PMID- 11034619 TI - Protein kinase C-dependent modulation of Na+ currents increases the excitability of rat neocortical pyramidal neurones. AB - The effect of the protein kinase C (PKC) activator 1-oleoyl-2-acetyl-sn-glycerol (OAG) on TTX-sensitive Na+ currents in neocortical pyramidal neurones was evaluated using voltage-clamp and intracellular current-clamp recordings. In pyramid-shaped dissociated neurones, the addition of OAG to the superfusing medium consistently led to a 30% reduction in the maximal peak amplitude of the transient sodium current (I(Na,T)) evoked from a holding potential of -70 mV. We attributed this inhibitory effect to a significant negative shift of the voltage dependence of steady-state channel inactivation (of approximately 14 mV). The inhibitory effect was completely prevented by hyperpolarising prepulses to potentials that were more negative than -80 mV. A small but significant leftward shift of INa,T activation was also observed, resulting in a slight increase of the currents evoked by test pulses at potentials more negative then -35 mV. In the presence of OAG, the activation of the persistent fraction of the Na+ current (INa,P) evoked by means of slow ramp depolarisations was consistently shifted in the negative direction by 3.9+/-0.5 mV, while the peak amplitude of the current was unaffected. In slice experiments, the OAG perfusion enhanced a subthreshold depolarising rectification affecting the membrane response to the injection of positive current pulses, and thus led the neurones to fire in response to significantly lower depolarising stimuli than those needed under control conditions. This effect was attributed to an OAG-induced enhancement of INa,P, since it was observed in the same range of potentials over which I(Na,P) activates and was completely abolished by TTX. The qualitative firing characteristics of both the intrinsically bursting and regular spiking neurones were unaffected when OAG was added to the physiological perfusing medium, but their firing frequency increased in response to slight suprathreshold depolarisations. The obtained results suggest that physiopathological events working through PKC activation can increase neuronal excitability by directly amplifying the I(Na,P)-dependent subthreshold depolarisation, and that this facilitating effect may override the expected reduction in neuronal excitability deriving from OAG-induced inhibition of the maximal INa, T peak amplitude. PMID- 11034620 TI - CO2 permeability and bicarbonate transport in microperfused interlobular ducts isolated from guinea-pig pancreas. AB - Permeabilities of the luminal and basolateral membranes of pancreatic duct cells to CO2 and HCO3- were examined in interlobular duct segments isolated from guinea pig pancreas. Intracellular pH (pHi) was measured by microfluorometry in unstimulated, microperfused ducts loaded with the pH-sensitive fluoroprobe 2'7' bis(2-carboxyethyl)-5(6)-carboxyfluorescein (BCECF). When HCO3-/CO2 was admitted to the bath, pHi decreased transiently as a result of CO2 diffusion and then increased to a higher value as a result of HCO3- uptake across the basolateral membrane by Na+-HCO3- cotransport. When HCO3-/CO2 was admitted to the lumen, pHi again decreased but no subsequent increase was observed, indicating that the luminal membrane was permeable to CO2 but did not allow HCO3- entry to the cells from the lumen. Only when the luminal HCO3- concentration was raised above 125 mM was HCO3- entry detected. The same was true of duct cells stimulated with forskolin. Recovery of pHi from an acid load, induced by exposure to an NH4+ pulse, was dependent on basolateral but not luminal Na+ and could be blocked by basolateral application of methylisobutylamiloride and H2DIDS. This indicates that the Na+-H+ exchangers and Na+-HCO3- cotransporters are located exclusively at the basolateral membrane. In the presence of HCO3-/CO2, substitution of basolateral Cl- with glucuronate caused larger increases in pHi than substitution of luminal Cl-. This suggests that the anion exchanger activity in the basolateral membrane is greater than that in the luminal membrane. We conclude that the luminal and basolateral membranes are both freely permeable to CO2, but while the basolateral membrane has both uptake and efflux pathways for HCO3-, the luminal membrane presents a significant barrier to the re-entry of secreted HCO3 , largely through the inhibition of the luminal anion exchanger by high luminal HCO3- concentrations. PMID- 11034621 TI - Real-time studies of zymogen granule exocytosis in intact rat pancreatic acinar cells. AB - An adequate understanding of secretion requires the measurement of exocytosis on the same time scale as that used for second messenger dynamics. To investigate the kinetics of ACh-evoked secretion in pancreatic acinar cells, exocytosis of zymogen granules was quantified by continuous, time-differential analysis of digital images. The validity of this method was confirmed by simultaneous fluorescence imaging of quinacrine-loaded zymogen granules. Basal rates of exocytosis were low (0.2 events min(-1)). ACh stimulated a biphasic increase in secretory activity, maximal rates exceeding 20 events min(-1) after 10 s of ACh application (10 microM). Over the next 15 s the rate of exocytosis fell to less than 4 events min(-1); then began a second phase of secretion that peaked 15 s later at approximately 11 events min(-1), but subsequently declined in the continued presence of agonist. Measurements of fura-2 fluorescence demonstrated a biphasic increase in intracellular [Ca2+] ([Ca2+]i). Comparison of the [Ca2+]i records and time-differential analysis revealed that the fall in exocytotic rate following the initial burst occurred despite the fact that [Ca2+]i remained high. The second phase of secretion depended on both [Ca2+]i and [ACh]. At 10 microM ACh there was a decrease in the steepness of the relationship between [Ca2+]i and exocytosis that led to an enhancement of the slow secretory phase. We propose that acinar cells contain two pools of secretory vesicles: a small pool of granules that is exocytosed rapidly, but is quickly depleted; and a reserve pool of granules that can be recruited by ACh in a process that is modulated by second messengers other than calcium. PMID- 11034622 TI - Activation of Ca2+--calmodulin kinase II induces desensitization by background light in dogfish retinal 'on' bipolar cells. AB - Retinal 'on' bipolar cells possess a metabotropic glutamate receptor (mGluR6) linked to the control of a G-protein and cGMP-activated channels which functions to generate high synaptic amplification of rod signals under dark-adapted conditions. Desensitization of 'on' bipolar cells is initiated by a rise in Ca2+ during background light too weak to adapt rod photoreceptors. Desensitization could also be elicited by raising intracellular Ca2+ above 1 microM. In order to investigate the mechanism of desensitization, whole-cell current responses to brief flashes and to steps of light were obtained from voltage-clamped 'on' bipolar cells in dark-adapted dogfish retinal slices. The inclusion of Ca2+ calmodulin kinase II (CaMKII) inhibitor peptides in the patch pipette solutions not only blocked desensitization of 'on' bipolar cells by dim background light and by 50 microM Ca2+, but also increased their flash sensitivity. The substrate of phosphorylation by CaMKII is the 'on' bipolar cell cGMP-activated channels. Desensitization probably results from a reduction in their sensitivity to cGMP and a voltage-dependent decrease in their conductance. A role for protein kinase C (PKC) in this process was excluded since activating PKC independently of Ca2+ with the phorbol ester PMA failed to induce desensitization of 'on' bipolar cells. PMID- 11034623 TI - Activation of silent mechanoreceptive cat C and Adelta sensory neurons and their substance P expression following peripheral inflammation. AB - The effect of inflammation on the excitability and the level of substance P (SP) in cat mechanoreceptive C and Adelta dorsal root ganglion (DRG) neurons were studied in vivo using intracellular recording and immunocytochemical techniques. Following injections of carrageenan (Carg) into the cat hindpaw, the percentage of C neurons exhibiting spontaneous activity increased from 7.2 to 20.7% and the percentage of Adelta neurons increased from 6.9 to 18.6%. In contrast to most cells from normal cats, which fired regularly below 10 Hz, many cells from Carg treated cats fired at higher frequencies or in bursts. Inflammation (Carg treatment) also depolarized membrane potentials, increased membrane input resistance, caused the disappearance of inward rectifying currents and lowered the mean current thresholds of tibial nerve-evoked responses in DRG neurons. With inflammation, the percentage of C or Adelta neurons responding to low threshold mechanoreceptive stimuli increased (C neurons: normal, 13%; inflamed, 41%; Adelta neurons: normal, 13 %; inflamed, 39 %), while the percentage of C or Adelta neurons responding to high threshold mechanoreceptive stimuli remained unchanged. Some receptive field (RF)-responsive cells were injected with Lucifer Yellow and their SP immunoreactivity was determined. Following Carg treatment, substantially higher percentages of RF-responsive cells were SP positive (C neurons: normal, 35.7%; inflamed, 60%; Adelta neurons: normal, 18.2%; inflamed, 66.7%). These combined increases in the excitability of DRG neurons and SP-containing RF responsive neurons could lead to sensitization of sensory neurons, thus contributing to the development of hyperalgesia. PMID- 11034624 TI - Reflex effects of independent stimulation of coronary and left ventricular mechanoreceptors in anaesthetised dogs. AB - Previous studies which have indicated that the stimulation of ventricular mechanoreceptors induces significant reflex responses can be criticised because of the likelihood of concomitant stimulation of coronary arterial baroreceptors. We therefore undertook this investigation to examine the coronary and ventricular mechanoreflexes in a preparation in which the pressure stimuli to each region were effectively separated. Dogs were anaesthetised, artificially ventilated and placed on cardiopulmonary bypass. A balloon at the ventricular outflow separated pressure in the left ventricle from that perfusing the coronary arteries. Ventricular pressures were changed by varying inflow and outflow of blood entering and leaving the ventricle through an apical cannula, and coronary pressure by changing pressure in a reservoir connected to a cannula tied in the aortic root. Pressures distending carotid and aortic baroreceptors were controlled. Changes in descending aortic perfusion pressure (flow constant) were used to assess systemic vascular responses. Large changes in carotid sinus and coronary pressures decreased vascular resistance by 35+/-1.9 and 40+/-2.5%, respectively. Intracoronary injections of veratridine (30-60 microg) decreased vascular resistance by 31+/-2.5%. However, large increases in ventricular pressure decreased resistance by only 9+/-2.2%. Significant changes in vascular resistance were obtained with increases in coronary arterial pressure from 60 to 90 mmHg. However, ventricular pressures had to increase to 152/18 mmHg (systolic/end-diastolic) before there was a significant response. These results show that coronary mechanoreceptors are likely to play an important role in cardiovascular control. If ventricular receptors have any function at all, it is as a protective mechanism during gross distension, possibly associated with myocardial ischaemia. PMID- 11034625 TI - Intersample fluctuations in phosphocreatine concentration determined by 31P magnetic resonance spectroscopy and parameter estimation of metabolic responses to exercise in humans. AB - The ATP turnover rate during constant-load exercise is often estimated from the initial rate of change of phosphocreatine concentration ([PCr]) using 31P magnetic resonance spectroscopy (MRS). However, the phase and amplitude characteristics of the sample-to-sample fluctuations can markedly influence this estimation (as well as that for the time constant (tau) of the [PCr] change) and confound its physiological interpretation especially for small amplitude responses. This influence was investigated in six healthy males who performed repeated constant-load quadriceps exercise of a moderate intensity in a whole body MRS system. A transmit- receive surface coil was placed under the right quadriceps, allowing determination of intramuscular [PCr]; pulmonary oxygen uptake (VO2) was simultaneously determined, breath-by-breath, using a mass spectrometer and a turbine volume measuring module. The probability density functions (PDF) of [PCr] and VO2 fluctuations were determined for each test during the steady states of rest and exercise and the PDF was then fitted to a Gaussian function. The standard deviation of the [PCr] and VO2 fluctuations at rest and during exercise (sr and sw, respectively) and the peak centres of the distributions (xc(r) and xc(w)) were determined, as were the skewness (gamma1) and kurtosis (gamma2) coefficients. There was no difference between sr and sw for [PCr] relative to the resting control baseline (s(r) = 1.554 %delta (s.d. = 0.44), s(w) = 1.514 %delta (s.d. = 0.35)) or the PDF peak centres (xc(r) = -0.013 %delta (s.d. = 0.09), xc(w) -0.197 %delta (s.d. = 0.18)). The standard deviation and peak centre of the 'noise' in VO2 also did not vary between rest and exercise (sr = 0.0427 l min(-1) (s.d. = 0.0104), s(w) = 0.0640 l min(-1) (s.d. = 0.0292); xc(r) = -0.0051 l min(-1) (s.d. = 0.0069), xc(w) 0.0022 l min(-1) (s.d. = 0.0034)). Our results demonstrate that the intersample 'noise' associated with [PCr] determination by 31P-MRS may be characterised as a stochastic Gaussian process that is uncorrelated with work rate, as previously described for VO2. This 'noise' can significantly affect the estimation of tau[PCr] and especially the initial rate of change of [PCr], i.e. the fluctuations can lead to variations in estimation of the initial rate of change of [PCr] of more than twofold, if the inherent 'noise' is not accounted for. This 'error' may be significantly reduced in such cases if the initial rate of change is estimated from the time constant and amplitude of the response. PMID- 11034626 TI - Adrenaline and glycogenolysis in skeletal muscle during exercise: a study in adrenalectomised humans. AB - The role of adrenaline in regulating muscle glycogenolysis and hormone-sensitive lipase (HSL) activity during exercise was examined in six adrenaline-deficient bilaterally adrenalectomised, adrenocortico-hormonal-substituted humans (Adr) and in six healthy control individuals (Con). Subjects cycled for 45 min at approximately 70% maximal pulmonary O2 uptake (VO2,max) followed by 15 min at approximately 86% VO2,max either without (-Adr and Con) or with (+Adr) adrenaline infusion that elevated plasma adrenaline levels (45 min, 4.49+/-0.69 nmol l(-1); 60 min, 12.41+/-1.80 nmol l(-1)). Muscle samples were obtained at 0, 45 and 60 min of exercise. In -Adr and Con, muscle glycogen was similar at rest (-Adr, 409+/-19 mmol (kg dry wt)(-1); Con, 453+/-24 mmol (kg dry wt)(-1)) and following exercise (-Adr, 237+/-52 mmol (kg dry wt)(-1); Con, 227+/-50 mmol (kg dry wt)( 1)). Muscle lactate, glucose-6-phosphate and glucose were similar in -Adr and Con, whereas glycogen phosphorylase (a/a + b x 100 %) and HSL (% phosphorylated) activities increased during exercise in Con only. Adrenaline infusion increased activities of phosphorylase and HSL as well as blood lactate concentrations compared with those in -Adr, but did not enhance glycogen breakdown (+Adr, glycogen following exercise: 274+/-55 mmol (kg dry wt)(-1)) in contracting muscle. The present findings demonstrate that during exercise muscle glycogenolysis can occur in the absence of adrenaline, and that adrenaline does not enhance muscle glycogenolysis in exercising adrenalectomised subjects. Although adrenaline increases the glycogen phosphorylase activity it is not essential for glycogen breakdown in contracting muscle. Finally, a novel finding is that the activity of HSL in human muscle is increased in exercising man and this is due, at least partly, to stimulation by adrenaline. PMID- 11034627 TI - Mitochondrial function and antioxidative defence in human muscle: effects of endurance training and oxidative stress. AB - The influence of endurance training on oxidative phosphorylation and the susceptibility of mitochondrial oxidative function to reactive oxygen species (ROS) was investigated in skeletal muscle of four men and four women. Mitochondria were isolated from muscle biopsies taken before and after 6 weeks of endurance training. Mitochondrial respiration was measured before and after exposure of mitochondria to exogenous ROS (H2O2 + FeCl2). Endurance training increased peak pulmonary O2 uptake (VO2,peak) by 24 % and maximal ADP-stimulated mitochondrial oxygen consumption (state 3) by 40% (P<0.05). Respiration in the absence of ADP (state 4), the respiratory control ratio (RCR = state 3/state 4) and the ratio between added ADP and consumed oxygen (P/O) remained unchanged by the training programme. Exposure to ROS reduced state 3 respiration but the effect was not significantly different between pre- and post-training samples. State 4 oxygen consumption increased after exposure to ROS both before (+189 %, P< 0.05) and after training (+243 %, P<0.05) and the effect was significantly higher after training (P<0.05, pre- vs. post-training). The augmented state 4 respiration could in part be attenuated by atractyloside, which indicates that ADP/ATP translocase was affected by ROS. The P/O ratio in ROS-treated mitochondria was significantly lower (P<0.05) compared to control conditions, both before (-18.6+/-2.2 %) and after training (-18.5+/-1.1%). Muscle activities of superoxide dismutase (mitochondrial and cytosolic), glutathione peroxidase and muscle glutathione status were unaffected by training. There was a positive correlation between muscle superoxide dismutase activity and age (r = 0.75; P<0.05; range of age 20-37 years), which may reflect an adaptation to increased generation of ROS in senescent muscle. The muscle glutathione pool was more reduced in subjects with high activity of glutathione peroxidase (r = 0.81; P<0.05). The influence of short-term training on mitochondrial oxygen consumption has for the first time been investigated in human skeletal muscle. The results showed that maximal mitochondrial oxidative power is increased after endurance training but that the efficiency of energy transfer (P/O ratio) remained unchanged. Antioxidative defence was unchanged after training when expressed relative to muscle weight. Although this corresponds to a reduced antioxidant protection per individual mitochondrion, the sensitivity of aerobic energy transfer to ROS was unchanged. However, the augmented ROS-induced non-coupled respiration after training indicates an increased susceptibility of mitochondrial membrane proton conductance to oxidative stress. PMID- 11034628 TI - The initiation of the swing phase in human infant stepping: importance of hip position and leg loading. AB - Hip extension and low load in the extensor muscles are important sensory signals that allow a decerebrate or spinal cat to advance from the stance phase to the swing phase during walking. We tested whether the same sensory information controlled the phases of stepping in human infants. Twenty-two infants between the ages of 5 and 12 months were studied during supported stepping on a treadmill. Forces exerted by the lower limbs, surface electromyography (EMG) from muscles, and the right hip angle were recorded. The whole experimental session was videotaped. The hip position and the amount of load experienced by the right limb were manipulated during stepping by changing the position of the foot during the stance phase or by applying manual pressure on the pelvic crest. Disturbances with different combinations of hip position and load were used. The stance phase was prolonged and the swing phase delayed when the hip was flexed and the load on the limb was high. In contrast, stance phase was shortened and swing advanced when the hip was extended and the load was low. The results were remarkably similar to those in reduced preparations of the cat. They thus suggest that the behaviour of the brainstem and spinal circuitry for walking may be similar between human infants and cats. There was an inverse relationship between hip position and load at the time of swing initiation, indicating the two factors combine to regulate the transition. PMID- 11034629 TI - Cancer-related symptoms. AB - Patients with cancer experience multiple symptoms including pain, dyspnea, fatigue, depression, and cognitive impairment. These symptoms impair patients' daily functioning and their quality of life. Symptoms that can be well managed are often undertreated. A major barrier to adequate symptom treatment is poor assessment. The use of simple measurement scales greatly improves the symptom assessment process, helps direct treatment choices, and provides information about the effectiveness of treatment. Recently, better methods for symptom assessment have been developed, including brief self-report tools for the assessment of multiple symptoms and interactive voice response systems for assessing symptoms at home. Symptom assessment can be linked to evidence-based or best practice guidelines to expedite optimal symptom treatment. Because patients with cancer receiving radiotherapy are seen in the clinic frequently, the radiation oncologist can play an integral role in a comprehensive approach that involves both the medical and radiotherapeutic treatment of cancer-related symptoms. PMID- 11034630 TI - Fractionation and outcomes with palliative radiation therapy. AB - A dose-response relationship can be established for local control of a variety of malignancies treated with radiation, yet palliation of symptoms oftentimes does not have a clear dose-response relationship. It is important that palliation be achieved with as efficient a fractionation schedule as possible in patients with limited life expectancy and with as few side effects as possible. This article reviews the literature addressing optimal schedules of radiation for palliation based on prognostic factors. PMID- 11034631 TI - Tissue tolerance to reirradiation. AB - There are increasing requests for delivering a second course of radiation to patients who develop second primary tumors within or close to previous radiotherapy portal or late in-field recurrences. Rational treatment decisions demand rather precise knowledge on long-term recovery of occult radiation injury in various organs. This article summarizes available experimental and clinical data on the effects of reirradiation to the skin, mucosa, gut, lung, spinal cord, brain, heart, bladder, and kidney. The data reveal that, in general, acutely responding tissues recover radiation injury within a few months and, therefore, can tolerate another full course of radiation. For late toxicity endpoints, however, tissues vary considerably in their capacity to recover from occult radiation damage. The heart, bladder, and kidney do not exhibit long-term recovery at all. In contrast, the skin, mucosa, lung, and spinal cord do recover subclinical injury partially to a magnitude dependent on the organ type, size of the initial dose, and, to a lesser extent, the interval between radiation courses. The available clinical data have inspired many radiation oncologists to undertake systematic studies addressing the efficacy and toxicity of reirradiation in various clinical settings. Hopefully, systematic scoring, collection, and analysis of patient outcome will produce quantitative data useful for clinical practice. PMID- 11034632 TI - Clinical experience with retreatment for palliation. AB - Every radiation oncologist is faced occasionally with the need to consider reirradiation for palliation. Because reirradiation has the potential to exceed normal tissue tolerances, there is a need to have information on the efficacy and toxicity of retreatment. This article reviews the reirradiation literature and provides guidance to clinicians with regard to the risks, benefits, and side effects of retreatment. PMID- 11034633 TI - The role of brachytherapy for palliation. AB - The goal of palliative radiation is to alleviate symptoms in a short amount of time and maintain an optimal functional and quality-of-life level while minimizing toxicity and patient inconvenience. Despite advances in multimodality antineoplastic therapies, failure to control the tumor at its primary site frustratingly remains the predominant source of morbidity and mortality in many patients with cancer. Escalation of doses of radiation using external beam irradiation has been shown to improve local tumor control, but limits are imposed by the tolerance of normal surrounding structures. The highly conformal nature of brachytherapy enables the radiation oncologist to accomplish safe escalation of radiation doses to the tumor while minimizing doses to normal surrounding structures. Thus, by enhancing the potential for local control, brachytherapy used alone or as a supplement to external beam radiation therapy retains a significant and important role in achieving the goals of palliation. Proper patient selection, excellent technique, and adherence to implant rules will minimize the risk of complications. The advantages realized with the use of brachytherapy include good patient tolerance, short treatment time, and high rates of sustained palliation. This article reviews various aspects of palliative brachytherapy, including patient selection criteria, implant techniques, treatment planning, dose and fractionation schedules, results, and complications of treatment. Tumors of the head and neck, trachea and bronchi, esophagus, biliary tract, and brain, all in which local failure represents the predominant cause of morbidity and mortality, are highlighted. PMID- 11034634 TI - Systemic radiopharmaceutical therapy of painful osteoblastic metastases. AB - Bone pain from osteoblastic metastases can be ameliorated 40% to 80% of the time. Although we can predict nonresponders, we cannot predict responders; however, patients with a better performance scale may have a better chance of pain relief. Radiopharmaceuticals containing phosphorus 32, strontium 89, samarium 153, rhenium 186, and tin 117m are effective, but we do not know which is the most efficacious and the safest. Toxicity includes the flare phenomenon and mild to moderate pancytopenia, but disseminated intravascular coagulation can cause severe, life-threatening thrombocytopenia. This treatment may be repeated at about 9- to 12-week intervals, perhaps earlier with (153)Sm lexidronam, (186)Re etidronate, and (117m)Sn pentetate, with a success rate approaching that of the initial injection. The duration of action of pain reduction ranges from 2 weeks to many months. Tumorical effects are probably not the only mechanism of pain relief. PMID- 11034635 TI - The role of bisphosphonates in metastatic breast cancer. AB - The American Society of Clinical Oncology has recently developed guidelines for the use of bisphosphonates in breast cancer. Highlights of these guidelines are reviewed. Specific issues addressed included when in the course of disease should treatment be started, the duration of therapy, the role of bisphosphonates in pain control, their safety, and current estimates of their cost effectiveness. Although intravenous bisphosphonates, primarily pamidronate, have been shown to reduce the frequency of skeletal-related complications in women with known lytic bone metastases from breast cancer, numerous major areas of clinical importance related to their use have not been studied and are highlighted. PMID- 11034636 TI - Future strategies needed for palliative care. AB - Too many cancer patients have less than optimal care at the end of life, as measured by unrelieved pain, death in a setting other than home, and uncoordinated care. Solutions will require at least the following: (1) new models of care and better coordination of care; (2) new drugs or techniques; (3) better professional knowledge and dissemination of that knowledge; (4) facing the issue of death; and (5) acknowledgment of cost constraints. Cost constraints will lead to more gaps between the haves and have nots in the United States in the next 5 years. PMID- 11034637 TI - 1999 North American Spine Society Presidential Address. The millennium threshold: is it the economy, stupid? PMID- 11034638 TI - Depression and chronic low back pain: establishing priorities in treatment. PMID- 11034639 TI - The biopsychosocial approach to spine care and research. PMID- 11034640 TI - Human intervertebral disc cells are genetically modifiable by adenovirus-mediated gene transfer: implications for the clinical management of intervertebral disc disorders. AB - STUDY DESIGN: Human intervertebral disc cells were cultured in monolayer and treated with adenovirus-containing marker genes to determine the susceptibility of the cells to adenovirus-mediated gene transfer. OBJECTIVES: To test the efficacy of the adenovirus-mediated gene transfer technique for transferring exogenous genes to human intervertebral disc cells in vitro. SUMMARY OF BACKGROUND DATA: Upregulated proteoglycan synthesis after direct in vivo adenovirus-mediated transfer of growth factor genes to the rabbit intervertebral disc has previously been reported. Before contemplating extending this approach to the treatment of human disc disease, it is necessary to demonstrate that human intervertebral disc cells are indeed susceptible to adenovirus-mediated gene transduction. METHODS: Human intervertebral disc cells were isolated from disc tissue obtained from 15 patients during surgical disc procedures. The cells were cultured in monolayer and treated with saline containing five different doses of adenovirus carrying the lacZ gene (Ad/CMV-lacZ), saline containing adenovirus carrying the luciferase gene (Ad/CMV-luciferase), or saline alone. Transgene expression was analyzed by 5-bromo-4-chloro-3-indolyl-beta-galactosidase (X-Gal) staining and luciferase assay. RESULTS: Adenovirus efficiently transferred lacZ and luciferase marker genes to cells from degenerated discs as well as to cells from nondegenerated discs. A minimum dose of 150 MOI Ad/CMV-lacZ was found to be sufficient to achieve transduction of approximately 100% of disc cells-regardless of patient age, sex, surgical indication, disc level, and degeneration grade. No statistically significant difference in the luciferase activities could be detected in disc cell cultures from degenerated and nondegenerated discs treated with Ad/CMV-luciferase. CONCLUSIONS: In vitro transducibility of human intervertebral disc cells by adenovirus is relatively insensitive to disc degeneration grade. Because the rate-limiting step for successful gene therapy is the ability to transfer genes efficiently to the target tissue, the achievement of efficient gene transfer to human intervertebral disc cells(using a direct, adenovirus-mediated approach) is an important and necessary step in the development of gene therapy strategies for the management of human intervertebral disc disorders. PMID- 11034641 TI - Direct current electrical stimulation increases the fusion rate of spinal fusion cages. AB - STUDY DESIGN: A randomized experimental evaluation of direct current stimulation in a validated animal model with an experimental control group, using blinded radiographic, biomechanical, histologic, and statistical measures. OBJECTIVES: To evaluate the efficacy of the adjunctive use of direct current stimulation on the fusion rate and speed of healing of titanium interbody fusion cages packed with autograft in a sheep lumbar interbody fusion model. SUMMARY OF BACKGROUND DATA: Titanium lumbar interbody spinal fusion cages have been reported to be 90% effective for single-level lumbar interbody fusion. However, fusion rates are reported to be between 70% and 80% in patients with multilevel fusions or with risk factors such as obesity, tobacco use, or metabolic disorders. The authors hypothesized that direct current stimulation would increase the fusion rate of titanium interbody fusion cages packed with autograft in a sheep lumbar interbody fusion model. METHODS: Twenty-two sheep underwent lumbar discectomy and fusion at L4-L5 with an 11- x 20-mm Bagby and Kuslich (BAK) cage packed with autograft. Seven sheep received a BAK cage and no current. Seven sheep had a cage and a 40 microA current applied with a direct current stimulator. Eight sheep had a BAK cage and a 100-microA current applied. All sheep were killed 4 months after surgery. The efficacy of electrical stimulation in promoting interbody fusion was assessed by performing radiographic, biomechanical, and histologic analyses in a blinded fashion. RESULTS: The histologic fusion rate increased as the direct current dose increased from 0 microA to 40 microA to 100 microA (P < 0.009). Histologically, all animals in the 100-microA group had fusions in both the right and left sides of the cage. Direct current stimulation had a significant effect on increasing the stiffness of the treated motion segment in right lateral bending (P < 0.120), left lateral bending (P < 0.017), right axial rotation (P < 0.004), left axial rotation (P < 0.073), extension (P < 0.078), and flexion (P < 0.029) over nonstimulated levels. CONCLUSION: Direct current stimulation increased the histologic and biomechanical fusion rate and the speed of healing of lumbar interbody spinal fusion cages in an ovine model at 4 months. PMID- 11034642 TI - The effect of nicotine on gene expression during spine fusion. AB - STUDY DESIGN: A rabbit model of posterolateral spine fusion was used to investigate the effect of nicotine on cytokine expression during spine fusion. OBJECTIVES: To determine the effects of nicotine on the known gene expression pattern of bone morphogens and related proteins expressed during spine fusion. SUMMARY OF BACKGROUND DATA: The mechanism by which nicotine increases the pseudarthrosis rate of spine fusion is unknown. Recently, a distinct temporal and spatial pattern of cytokine expression during bone formation has been described. The authors hypothesized that nicotine would alter this known pattern, thereby revealing the mechanism by which nicotine exerts its effect. METHODS: Twenty eight New Zealand White rabbits underwent posterolateral spine fusion with autogenous bone graft. Fourteen rabbits received systemic nicotine by a miniosmotic pump. Fusions were harvested at 0, 2, 5, and 7 days and 2, 3, and 4 weeks after arthrodesis. Specimens were divided into the outer zones adjacent to the transverse processes and the central zones between the transverse processes. Gene expression of type I and II collagen, bone morphogenic protein-2, -4, and -6 and basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) was then measured at each time point in each of the two zones. RESULTS: Nicotine inhibited expression of all cytokines measured, mainly in the central zone. However, the previously described temporal and spatial patterns of expression were preserved. CONCLUSIONS: Nicotine inhibits expression of a wide range of cytokines, including those associated with neovascularization and osteoblast differentiation. Therefore, the effects of nicotine appear to involve more than just local vasoconstriction. PMID- 11034643 TI - Real-time in vivo loading in the lumbar spine: part 1. Interbody implant: load cell design and preliminary results. AB - STUDY DESIGN: Instrumented interbody implants were placed into the disc space of a motion segment in two baboons. During the animal's activities, implants directly measured in vivo loads in the lumbar spine by telemetry transmitter. OBJECTIVES: Develop and test an interbody implant-load cell and use the implant to measure directly loads imposed on the lumbar spine of the baboon, a semiupright animal. SUMMARY OF BACKGROUND DATA: In vivo forces in the lumbar spine have been estimated using body weight calculations, moment arm models, dynamic chain models, electromyogram measurements, and intervertebral disc pressure measurements. METHODS: An analytical model was used to determine the force-strain relation in a customized interbody implant. After validation by finite element modeling, strain gauges were mounted onto the implant and connected to a telemetry transmitter. Implants were placed surgically into the L4 L5 disc space of skeletally mature baboons and the transmitter in the flank. After surgery, load data were collected from the animals during activities. Radiographs were taken monthly to assess fusion. RESULTS: The implant-load cell is sufficiently sensitive to monitor dynamic changes in strain and load. During extreme activity, highest measurable strain values were indicative of loads in excess of 2.8 times body weight. CONCLUSIONS: The study technique and technology are efficacious for measuring real-time in vivo loads in the spine. Measuring load on an intradiscal implant over the course of healing provides key information about the mechanics of this process. Loads may be used to indicate performance demands on the intervertebral disc and interbody implants for subsequent implant design. PMID- 11034644 TI - Twelve-month follow-up of a controlled trial of intradiscal thermal anuloplasty for back pain due to internal disc disruption. AB - STUDY DESIGN: Case-control study. OBJECTIVES: To determine the prima facie efficacy of intradiscal electrothermal anuloplasty (IDTA). SUMMARY OF BACKGROUND DATA: Although it is being used increasingly as a putative treatment for internal disc disruption, no studies have been published on the efficacy of IDTA. METHODS: Fifty-three patients with back pain determined by computed tomographic (CT) discography to be due to internal disc disruption were offered treatment. The outcomes of 35 patients treated with IDTA were compared with those of a convenience sample of 17 patients treated with a physical rehabilitation program, by using a visual analog pain scale, use of analgesics, and return to work as measures. RESULTS: At 3 months, only one control patient obtained any significant degree of relief of pain, compared with 23 in the index group. Relief of pain was sustained at 6 and 12 months and was associated with improvement in disability, reduced drug use, and a return to work rate of 53%. Depending on the stringency of criteria used, the success rate of IDTA may be as low as 23% or as high as 60% with confidence intervals of +/-16%. CONCLUSIONS: In carefully selected cases, IDTA can eliminate or dramatically reduce the pain of internal disc disruption in a substantial proportion of patients and appears to be superior to conventional conservative care for internal disc disruption. PMID- 11034645 TI - The effect of cigarette smoking and smoking cessation on spinal fusion. AB - STUDY DESIGN: The effect of cigarette smoking and smoking cessation on spinal fusion was studied in a retrospective review of 357 patients who had undergone instrumented spinal fusion. OBJECTIVE: To document the widely assumed but unreported benefit of cigarette smoking cessation on fusion rate and clinical outcome after spinal fusion surgery. BACKGROUND DATA: Cigarette smoking has been shown to inhibit lumbar spinal fusion and to adversely effect outcome in treatment of lumbar spinal disorders. Prior reports have compared smokers and nonsmokers, as opposed to comparing smokers and quitters. METHODS: This study retrospectively identified 357 patients who underwent a posterior instrumented fusion at either L4-L5 or L4-S1 between 1992 and 1996. Analysis of the medical record and follow-up telephone surveys were conducted. Clinical outcome and fusion status was analyzed in relation to preoperative and postoperative smoking parameters. RESULTS: In this study, the nonunion rate was 14.2% for nonsmokers and 26.5% for patients who continued to smoke after surgery (P < 0.05). Patients who quit smoking after surgery for longer than 6 months had a nonunion rate of 17.1%. The nonunion rate was not significantly affected by either the quantity that a patient smoked before surgery or the duration of preoperative smoking abatement. Return-to-work was achieved in 71% of nonsmokers, 53% of nonquitters, and 75% of patients who quit smoking for more than 6 months after surgery. DISCUSSION: These results validate the hypothetical assumption that postoperative smoking cessation helps to reverse the impact of cigarette smoking on outcome after spinal fusion. PMID- 11034646 TI - A prospective study of psychological predictors of lumbar surgery outcome. AB - STUDY DESIGN: Prospective design in which 102 patients were evaluated with a battery of psychological assessment tests 1-2 weeks before surgery, and outcome was assessed 6 months and 1 year after surgery. OBJECTIVES: The study examined whether three aspects of psychological distress (depression, anxiety, and hostility) predict several surgical outcomes (employment status, subjective pain change ratings, and changes in functional abilities). SUMMARY OF BACKGROUND DATA: Surgery for back pain has been shown to yield poor results in 15-45% of patients. Tools are needed to identify those "at risk" for poor outcome. Aspects of emotional distress, including anxiety, depression, and hostility, have been found to be relevant to various illness outcomes (e.g., cancer, heart disease), but their influence has not been prospectively evaluated for back pain surgical outcome. METHODS: Study patients completed measures of distress before surgery, including the Spielberger Trait Anxiety Inventory, Zung Depression Scale, Modified Somatic Perception Questionnaire, and Cook-Medley Hostility Scale. At 1 year follow-up, patients completed pain change ratings, functional abilities measure (Dallas Pain Questionnaire), and questions about employment status. RESULTS: Multivariate regression analyses, controlling for significant demographic variables, found that failure to return to work was predicted by presurgical anxiety (P < 0.001) and depression (P < 0. 01); failure to report improvement in pain was predicted by presurgical somatic anxiety (P < 0.01) and depression (P < 0.058); and failure to report improved functional abilities was predicted by presurgical somatic anxiety (P < 0.01) and depression (P < 0.05). Hostility did not predict any outcome. Regression analyses found a strong predictor to be a combination of the Zung Depression Scale and Modified Somatic Perception Questionnaire, known as the Distress and Risk Assessment Method (DRAM). CONCLUSIONS: These results indicate that screening for presurgical distress is likely to identify those patients at risk for poor outcome. Studies to evaluate whether presurgical psychological treatment improves outcome are warranted. PMID- 11034647 TI - Intradiscal electrothermal treatment for chronic discogenic low back pain: a prospective outcome study with minimum 1-year follow-up. AB - STUDY DESIGN: Prospective case series. OBJECTIVE: To determine the outcome of patients with chronic low back pain whose symptoms did not improve with aggressive nonoperative care and who chose (intradiscal electrothermal anuloplasty) IDET as an alternative to chronic pain management or interbody fusion surgery. SUMMARY OF BACKGROUND DATA: Patients with unremitting chronic discogenic low back pain are faced with the choice of long-term pain management or fusion surgery. Intradiscal electrothermal anuloplasty (IDET) was developed as an alternative minimally invasive treatment. MATERIALS AND METHODS: Sixty-two patients from the author's practice who had chronic low back pain unresponsive to nonoperative care, no evidence of compressive radiculopathy, and concordant pain reproduction at one or more disc levels on provocative discography were enrolled in the study. Visual analog scale (VAS) pain scores and Short Form (SF)-36 Health Status Questionnaire Physical Function subscale and SF-36 Bodily Pain subscale scores were assessed at baseline and at least 1 year later. RESULTS: Mean follow up was 16 months, and mean preoperative duration of symptoms was 60 months. Baseline and follow-up outcome measures demonstrated a mean change in VAS score of 3.0 (P < 0.001), mean change in SF-36 physical function of 20 (P < 0.001), and mean change in SF-36 bodily pain of 17 (P < 0.001). Symptoms improved in 44 (71%) of 62 of the study group on the SF-36 physical function subscale, in 46 (74%) of 62 on the SF-36 Bodily Pain subscale, and in 44 (71%) of 62 on the VAS scores. Twelve (19%) of 62 of the patients did not show improvement on any scale. CONCLUSION: A cohort of patients with chronic unremitting low back pain of discogenic origin whose symptoms had failed to improve with aggressive nonoperative care demonstrated a statistically significant and clinically meaningful improvement on the SF-36 and the VAS scores at a minimum follow-up of 1 year after IDET. The positive results should be validated with placebo controlled randomized trials and studies that compare IDET with alternative treatments.- PMID- 11034648 TI - Incidence of intravascular penetration in transforaminal lumbosacral epidural steroid injections. AB - STUDY DESIGN: A prospective, observational, human, in vivo study. OBJECTIVES: To evaluate the incidence of vascular penetration during fluoroscopically guided, contrast-enhanced, transforaminal lumbar epidural steroid injections (ESIs) and determine whether a "flash" (blood in the needle hub) or aspiration of blood can be used to predict a vascular injection. SUMMARY OF BACKGROUND DATA: Incorrectly placed, intravascular lumbosacral spinal injections result in systemic medication flow that misses the desired target. No previous studies evaluate the incidence of vascular injections in transforaminal ESIs, nor the ability of flash to predict a vascular injection. METHODS: The incidence of flash or positive blood aspiration and the incidence of fluoroscopically confirmed vascular spread were prospectively observed in 670 patients treated with lumbosacral fluoroscopically guided transforaminal ESIs. Presence of a flash or positive aspiration was documented. Contrast was injected to determine whether the needle tip was intravascular. RESULTS: Seven hundred sixty-one transforaminal ESIs were included. The overall rate of intravascular injections was 11.2%. There was a statistically significant higher rate of intravascular injections (21.3%) noted with transforaminal ESIs performed at S1 (n = 178), compared with those at the lumbar levels (8.1%, n = 583). Using flash or positive blood aspirate to predict intravascular injections was 97.9% specific, but only 44.7% sensitive. CONCLUSIONS: There is a high incidence of intravascular injections in transforaminal ESIs that is significantly increased at S1. Using a flash or blood aspiration to predict an intravascular injection is not sensitive, and therefore a negative flash or aspiration is not reliable. Fluoroscopically guided procedures without contrast confirmation are instilling medications intravascularly and therefore not into the desired epidural location. This finding confirms the need for not only fluoroscopic guidance but also contrast injection instillation in lumbosacral transforaminal ESIs. PMID- 11034649 TI - Imaging pitfalls of interbody spinal implants. AB - STUDY DESIGN: A variety of interbody implants were imaged by computed tomography and plain radiography within cadaveric spines to evaluate their basic imaging characteristics. OBJECTIVES: Sources of interpretation error by both computed tomography and plain radiography of interbody implants were investigated. SUMMARY OF BACKGROUND DATA: Lucencies have been reported around bone dowel implants in the postoperative period, which have been shown to resolve. The diagnosis of fusion through metallic implants has been difficult with both false-positive and false-negative results. The literature of imaging these implants is very sparse. METHODS: Four interbody constructs were placed in cadaveric spines under different conditions and imaged using both computed tomography and plain film radiography. RESULTS: Plain radiographs could not predict the presence of intraimplant bone whereas computed tomography was accurate. Metallic implants had a 1-3-mm computed tomography artifact limiting peri-implant interpretation. Lucencies could be seen on computed tomography but not on plain radiographs. However, the opposite was also seen. Lucencies around nonmetallic implants were more visible by a 4:1 ratio. CONCLUSIONS: Potential errors in interpretation were identified including assessment for bridging bone, assessment for lucency, and obscuration of peri-implant detail from metallic artifact. Lucencies were more visible with nonmetallic implants than with metallic constructs. Plain radiograph analysis of metallic implants tended to underestimate lucencies, whereas analysis of nonmetallic implants tended to overestimate lucencies. PMID- 11034650 TI - Radiation exposure to the spine surgeon during fluoroscopically assisted pedicle screw insertion. AB - STUDY DESIGN: In vitro study to determine occupational radiation exposure during lumbar fluoroscopy. OBJECTIVES: To assess radiation exposure to the spine surgeon during fluoroscopically assisted thoracolumbar pedicle screw placement. SUMMARY OF BACKGROUND DATA: Occupational radiation exposure during a variety of fluoroscopically assisted musculoskeletal procedures has been previously evaluated. No prior study has assessed fluoroscopy-related radiation exposure to the spine surgeon. METHODS: Bilateral pedicle screw placement (T11-S1) was performed in six cadavers using lateral fluoroscopic imaging. Radiation dose rates to the surgeon's neck, torso, and dominant hand were measured with dosimeter badges and thermolucent dosimeter (TLD) rings. Radiation levels were also quantified at various distances from the dorsal lumbar surface using an ion chamber radiation survey meter. RESULTS: The mean dose rate to the neck was 8.3 mrem/min. The dose rate to the torso was greatest when the surgeon was positioned ipsilateral to the beam source (53.3 mrem/min, compared with 2.2 mrem/min on the contralateral side). The average hand dose rate was 58.2 mrem/min. A significant increase in hand dose rate was associated with placement of screws ipsilateral to the beam source (P = 0.0005) and larger specimens (P = 0.0007). Radiation levels significantly decreased as distance from the beam source and dorsal body surface increased. The greatest levels of radiation were noted on the side where the primary radiograph beam entered the cadaver. CONCLUSION: Fluoroscopically assisted thoracolumbar pedicle screw placement exposes the spine surgeon to significantly greater radiation levels than other, nonspinal musculoskeletal procedures that involve the use of a fluoroscope. In fact, dose rates are up to 10-12 times greater. Spine surgeons performing fluoroscopically assisted thoracolumbar procedures should monitor their annual radiation exposure. Measures to reduce radiation exposure and surgeon awareness of high-exposure body and hand positions are certainly called for. PMID- 11034651 TI - A prospective randomized multicenter clinical evaluation of an anterior cervical fusion cage. AB - STUDY DESIGN: A prospective, concurrently controlled, randomized, multicenter trial of an anterior Bagby and Kuslich cervical fusion cage (BAK/C; Sulzer Spine Tech, Minneapolis, MN) for treatment of degenerative disc disease of the cervical spine. OBJECTIVES: To report clinical results with maximum 24-month follow-up of fusions performed with the BAK/C fusion cage. SUMMARY OF BACKGROUND DATA: Threaded lumbar cages have been used during the past decade as a safe and effective surgical solution for chronic disabling low back pain. Threaded cages have now been developed for use in anterior cervical interbody fusions to obviate the need for allografts or autogenous bone grafting procedures while providing initial stability during the fusion process. METHODS: Patients with symptomatic cervical discogenic radiculopathy were treated with either anterior cervical discectomy with uninstrumented bone-only fusion (ACDF) or BAK/C fusion cage(s). Independent radiographic assessment of fusion was made and patient-based outcome was assessed by visual analog pain scale and a Short Form (SF)-36 Health Status Questionnaire. RESULTS: Data analysis included 344 patients at 1 year and 180 at 2 years. When the two cage groups (hydroxya, patite-coated or noncoated) were compared with the ACDF group, similar outcomes were noted for duration of surgery, hospital stay, improvements in neck pain and radicular pain in the affected limb, improvements in the SF-36 Physical Component subscale and Mental Component subscale, and the patients' perception of overall surgical outcome. Symptom improvements were maintained at 2 years. A greater percentage of patients with ACDF needed an iliac crest bone harvest than did BAK/C patients (67% vs.- 3%). Successful fusion for one-level procedures at 12 months was 97.9% for the BAK/C groups and 89.7% for the ACDF group (P < 0.05). The complication rate for the ACDF group was 20.4% compared with an overall complication rate of 11.8% with BAK/C. There was no difference in complications that necessitated a second operative procedure. CONCLUSIONS: These results demonstrate that outcomes after a cervical fusion procedure with a threaded cage are the same as those of a conventional uninstrumented bone-only anterior discectomy and fusion with a low risk of complications and rare need for autogenous bone graft harvest. PMID- 11034652 TI - Four-year follow-up results of lumbar spine arthrodesis using the Bagby and Kuslich lumbar fusion cage. AB - STUDY DESIGN: This was a prospective multicenter clinical trial of a lumbar interbody fusion cage with a minimum of 4 years' follow-up. OBJECTIVES: To determine whether the early positive clinical results in fusions with lumbar cages, such as the Bagby and Kuslich (BAK) cage, are maintained beyond 2 years. SUMMARY OF BACKGROUND DATA: Threaded cages have been used increasingly for the treatment of symptomatic degenerative intervertebral disc disease. Concerns about the long-term clinical outcomes of this procedure have been posed, particularly regarding bony fusion viability, revision rates, potential adjacent level disease, and late complications. METHODS: The study cohort was a 196-patient subset from a prospective investigational device exemption. In addition to early postoperative examinations, these patients were examined biannually with a minimum of 4 years' follow-up. Patient outcome was assessed by a 6-point scale that evaluated pain relief, and functional improvement was determined by changes in activities of daily living. Fusion rates and return to work were determined. Complications and secondary operations were reported and categorized as non device related or device related. RESULTS: The patient cohort with 4-year follow up represented 25.6% of the original study population eligible at that time. Overall, the largest percentage of pain relief and functional improvements occurred by 3 months, and these improvements were maintained at each follow-up. Overall fusion rate was 91.7% and 95. 1% at 2 and 4 years, respectively. In this cohort, 39.5% of patients were working or were able to work within 3 months of surgery. After 4 years, 62.7% of patients were gainfully employed or able to work. The late-occurring complication rate in this cohort was 13.8% (27/196). Complications necessitating a second operation occurred in 8.7% (17/196), whereas reoperations that were deemed device related were performed in 3.1% (6/196). CONCLUSIONS: This study indicates that the early positive benefits of interbody fusion cage procedures are maintained through 4 years with acceptably low morbidity. PMID- 11034653 TI - Incidental durotomy in spine surgery. AB - STUDY DESIGN: Retrospective review of a large series of patients who underwent spinal surgery at a single institution during a 10-year period. OBJECTIVES: To further clarify the frequency of incidental durotomy during spine surgery, its treatment, associated complications, and results of long-term clinical follow-up. SUMMARY OF BACKGROUND DATA: Incidental durotomy is a relatively common occurrence during spinal surgery. There remains significant concern about it despite reports of good associated clinical outcomes. There have been few large clinical series on the subject. METHODS: A retrospective review was conducted of clinical and surgical records and radiographic data for consecutive patients who underwent spinal surgery performed by the two senior surgeons from January 1989 through December 1998. RESULTS: A total of 2144 patients were reviewed, and 74 were found to have dural tears occurring during or before surgery. Incidental durotomy occurred at the time of surgery in 66 patients (3.1% overall incidence). Incidence varied according to the specific procedure performed but was highest in the group that underwent revision surgery. The incidence of clinically significant durotomies occurring during surgery but not identified at the time was 0.28%. All dural tears that occurred during surgery and were recognized (60 of 66) were repaired primarily. Pseudomeningoceles developed in five of the remaining six patients. All six patients had subsequent surgical repair of dural defects because of failure of conservative therapy. A mean follow-up of 22.4 months was available and showed good long-term clinical results for all patients. CONCLUSIONS: Incidental durotomy, if recognized and treated appropriately, does not lead to long-term sequelae. PMID- 11034655 TI - Point of view PMID- 11034654 TI - Modification of C1-C2 transarticular screw fixation by image-guided surgery. AB - STUDY DESIGN: This is a feasibility study of image-guided surgery for C1-C2 transarticular screw fixation comparing postoperative screw position in a nonrandomized prospective cohort with a historic control group in which fluoroscopic guidance was used alone. OBJECTIVES: To evaluate the potential benefits and disadvantages of image-guided surgery for C1-C2 screw placement. SUMMARY OF BACKGROUND DATA: C1-C2 transarticular screw fixation is biomechanically superior to other current surgical stabilization procedures. The original technique for C1-C2 screw placement relies on anatomic landmarks and intraoperative fluoroscopy. Screw misplacement or anatomic variations can result in vertebral artery injury. Image-guided surgery involves using computed tomography (CT) data to plan the optimal screw trajectory before surgery and then use this data to guide screw placement during the actual surgery. Promising results of this technique are reported in the literature, but no direct comparison between image-guided surgery and conventional surgical techniques has been previously reported. METHODS: The image-guided surgery group consisted of 37 prospective patients. The historic control group included 78 patients who had similar surgeries performed using only fluoroscopic guidance. For the image guided surgery group, subluxation was reduced by positioning at the time of CT examination. The CT data were transferred to a StealthStation (Sofamor-Danek, Memphis, TN) surgical planning and guidance computer system, and an optimal screw trajectory was determined for the right and left transarticular screws. After matching the surgical field to the virtual computer field, C2 was drilled according to the planned screw trajectory, and screws were placed. Plain radiographs and CT were used for postoperative evaluation of the image-guided surgery group. RESULTS: Image-guided surgery reduced but did not eliminate the risk of screw misplacement. Surgical time was not increased overall. CONCLUSIONS: Image-guided surgery is an effective tool for the achievement of correct screw placement in C1-C2 transarticular screw fixation procedures. The procedure remains technically demanding. PMID- 11034656 TI - Cervical pedicle screws: comparative accuracy of two insertion techniques. AB - STUDY DESIGN: Independently assessed radiographic and anatomic comparison of device implantation methods. OBJECTIVES: To compare the relative accuracy of two techniques of inserting cervical pedicle screws. SUMMARY OF BACKGROUND DATA: In an attempt to define the anatomic risks of cervical pedicle screw insertion, image-guided stereotactic technology was shown to be superior to some other methods in vitro.- Meanwhile, in vivo experience with Abumi's technique of screw insertion has had few clinically relevant instances of screw malposition. There has been no direct comparison between current image-guided technology and Abumi's fluoroscopically assisted technique. METHODS: The pedicles (C3-C7) of human cadaveric cervical spines were instrumented with 3.5-mm screws with either of two techniques. Cortical integrity and potential neurovascular injury were independently assessed by computed tomographic (CT) scans and anatomic dissection. A cortical breach was considered "critical" if the screw encroached on any vital structure. If any part of the screw violated the cortex of the pedicle but no vital structure was at risk for injury, the breach was classified as "noncritical." RESULTS: In Group I (StealthStation; Sofamor-Danek, Memphis, TN), 82% of screws were placed in the pedicle, and 18% had a critical breach. In Group II (Abumi technique), 88% of screws were placed in the pedicle, and 12% had a critical breach. No statistically significant differences were demonstrated between each group (P = 0.59). Regarding pedicle dimensions and safety of insertion, a critical pedicle diameter of 4.5 mm was determined to be the size below which a critical breach was likely, but above which there was a significantly greater likelihood for safe screw placement. The most common structure injured in each group was the vertebral artery. CONCLUSIONS: The use of a computer-assisted image guidance system did not enhance safety or accuracy in placing pedicle screws compared with Abumi's technique. Both techniques have a noteworthy risk of injuring a critical structure if inserted into the pedicles with a diameter of less than 4.5 mm. Under laboratory conditions, pedicles with a diameter of more than 4.5 mm have a significantly greater likelihood of being safely instrumented by either technique. These data indicate that cervical pedicle screw placement is feasible, but it should be reserved for selected circumstances with clear indications and in the presence of suitable pedicle morphology. PMID- 11034657 TI - A prospective comparison of surgical approach for anterior L4-L5 fusion: laparoscopic versus mini anterior lumbar interbody fusion. AB - STUDY DESIGN: A prospective comparison of 50 consecutive patients who underwent L4-L5 anterior lumbar interbody fusion (ALIF). OBJECTIVES: To compare surgical time, blood loss, time in hospital, complications and adequacy of exposure between laparoscopic and mini-ALIF surgical approaches for L4-L5 anterior spinal fusion. SUMMARY OF BACKGROUND DATA: Advances in minimally invasive laparoscopic techniques have resulted in many centers adopting the endoscopic approach to L5 S1 as routine. However, the endoscopic approach to L4-L5 can be much more difficult. A direct comparison of open and laparoscopic techniques of exposure has not been reported. METHODS: From 1995 through 1998, data were prospectively collected on a series of 50 consecutive patients who underwent L4-L5 anterior interbody fusion with a threaded device, by either a laparoscopic or an open mini ALIF approach. RESULTS: Twenty-five patients underwent a laparoscopic procedure and 25 an open mini-ALIF approach. For single-level L4-L5 fusions, there was no statistical difference in operating time, blood loss, or length of hospital stay between laparoscopic or mini-ALIF groups. For two-level procedures, only the operative time differed, with laparoscopic procedures taking 25 minutes longer (P = 0.035). The rate of complications was significantly higher in the laparoscopic group (20% vs. 4%). In the laparoscopic group, 16% of patients had inadequate exposure, with the result that only a single cage was placed. In the open mini ALIF group, two cages were placed in all cases. CONCLUSIONS: There does not appear to be a significant advantage at the L4-L5 level of the transperitoneal laparoscopic surgical approach when compared with an open mini-ALIF retroperitoneal technique. PMID- 11034658 TI - Behavioral treatment for chronic low back pain: a systematic review within the framework of the Cochrane Back Review Group. AB - STUDY DESIGN: A systematic review of randomized controlled trials. SUMMARY OF BACKGROUND DATA: The treatment of chronic low back pain is not primarily focused on removing an underlying organic disease but at the reduction of disability through the modification of environmental contingencies and cognitive processes. Behavioral interventions are commonly used in the treatment of chronic (disabling) low back pain. OBJECTIVES: To determine whether behavioral therapy is more effective than reference treatments for chronic nonspecific low back pain and which type of behavioral treatment is most effective. METHODS: The authors searched the Medline and PsychLit databases and the Cochrane Controlled Trials Register up to April 1999, and Embase up to September 1999. Also screened were references of identified randomized trials and relevant systematic reviews. Methodologic quality assessment and data extraction were performed independently by two reviewers. The magnitude of effect was assessed by computing a pooled effect size for each domain (i.e., behavioral outcomes, overall improvement, back pain-specific and generic functional status, return to work, and pain intensity) using the random effects model. RESULTS: Only six (25%) studies were high quality. There is strong evidence (level 1) that behavioral treatment has a moderate positive effect on pain intensity (pooled effect size 0.62; 95% confidence interval [CI] 0. 25, 0.98), and small positive effects on generic functional status (pooled effect size 0.35; 95% CI: 0.04, 0.74) and behavioral outcomes (pooled effect size 0.40; 95% CI: 0.10, 0.70) of patients with chronic low back pain when compared with waiting-list controls or no treatment. There is moderate evidence (level 2) that a addition of behavioral component to a usual treatment program for chronic low backpain has no positive short-term effect on generic functional status (pooled effect size 0.31; 95% CI: 0.01, 0.64), pain intensity (pooled effect size 0.03; 95% CI: 0.30,0.36), and behavioral outcomes (pooled effect size 0.19; 95% CI: 0.08, 0.45). CONCLUSIONS: Behavioral treatment seems to be an effective treatment for patients with chronic low back pain,but it is still unknown what type of patients benefit most from what type of behavioral treatment. PMID- 11034659 TI - Editorial on residencies and fellowships. PMID- 11034661 TI - Educational: guidelines for orthopaedic and neurosurgical spinal fellowship training PMID- 11034660 TI - Resident and fellowship guidelines: educational guidelines for resident training in spinal surgery. PMID- 11034662 TI - Procedural guidelines for fellowship training in spinal surgery PMID- 11034664 TI - Imagery PMID- 11034665 TI - North american spine society: 14th annual meeting october 20-23, 1999 PMID- 11034663 TI - Transoral decompression, anterior plate fixation, and posterior wire fusion for irreducible atlantoaxial kyphosis in rheumatoid arthritis. AB - STUDY DESIGN: Fifteen consecutive patients with irreducible atlantoaxial kyphosis caused by rheumatoid arthritis were treated by combined transoral odontoid resection, anterior plate fixation, and posterior wire fusion. OBJECTIVES: To investigate the clinical results of this new surgical procedure. SUMMARY AND BACKGROUND DATA: Irreducible atlantoaxial kyphosis in rheumatoid arthritis results from a destruction of the craniocervical joint ligaments and the anterior aspects of the lateral atlantoaxial joints. The development of a paradental synovial pannus and atlantoaxial joint impaction prevents reduction by conservative treatment, such as skull traction. Posterior surgical procedures for the treatment of the irreducible atlantoaxial kyphosis with spinal cord compression have been associated with high morbidity and mortality. METHODS: Fifteen consecutive patients were treated by transoral odontoid resection. The fixation was performed with anterior plating, according to the method of Harms in combination with posterior wire fusion according to Brooks. Before and after surgery, evaluation was performed using the parameters of pain (visual analog scale), range of motion, and subjective assessment of improvement and the Health Assessment Questionnaire. The neurologic deficit was defined according to the classifications proposed by Ranawat, Frankel, and Nurwick. Plain radiographs, including lateral flexion and extension views, and magnetic resonance scans were obtained. RESULTS: No perioperative fatality occurred. The average clinical and radiographic follow-up was 50.7 +/- 15.6 months (range, 26-77). Postoperative pain was relieved (mean pain score before surgery, 7.9 +/- 1.87; after surgery, 3.8 +/- 1.27), and the range of motion of all patients increased (mean 21.5 +/- 14.0 degrees for rotation; mean 17.2 +/- 5. 54 degrees for bending). The score on the Health Assessment Questionnaire increased in three patients, remained unchanged in three and decreased in six patients (three had died). All patients improved at least one Ranawat level after surgery, except a patient in Ranawat Class II, whose condition remained unchanged. All patients were satisfied with the procedure and reported subjective improvement. CONCLUSION: Transoral plate fixation combined with posterior wire fixation after transoral odontoid resectionis an effective, reliable, and safe procedure for the treatment of irreducible atlantoaxial kyphosis in rheumatoid arthritis. PMID- 11034666 TI - Interventions for promoting adherence to tuberculosis management. AB - BACKGROUND: Up to half the people with tuberculosis do not complete their treatment. Strategies to improve adherence to diagnostic and treatment regimens are therefore important. OBJECTIVES: To assess the effects of various interventions aimed at promoting adherence to anti-tuberculosis treatment and completion of TB diagnostic protocols. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, the Cochrane Infectious Diseases Group trials register, Medline, Embase, Lilacs and reference lists of articles. We contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of interventions to promote adherence with curative or preventive chemotherapy and diagnostic protocols for tuberculosis. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Fourteen trials were included. Reminder cards sent to defaulters, a combination package of a monetary incentive and health education and more supervision of clinic staff increased the number of people completing their tuberculosis treatment. Intensive counselling/education did not help in one study. Direct observation showed better clinical outcomes in one study, and no difference in another. Return to the clinic for reading of a tuberculin skin test was enhanced by monetary incentives, assistance by lay health workers, contracts and telephone prompts but not by health education. REVIEWER'S CONCLUSIONS: We have found evidence of benefit for a number of specific interventions to improve adherence to anti-tuberculous therapy and completion of diagnostic protocols. These should be implemented by health care providers where appropriate to local circumstances. Future studies in low income countries are a priority and should measure adherence and clinical outcomes. PMID- 11034667 TI - Antibiotics for preventing respiratory tract infections in adults receiving intensive care. AB - BACKGROUND: Pneumonia is an important cause of mortality in intensive care units. The incidence of pneumonia in such patients ranges between 7% and 40%, and the crude mortality from ventilator associated pneumonia may exceed 50%. Although not all deaths in patients with this form of pneumonia are directly attributable to infections, it has been shown to contribute to mortality in intensive care units independently of other factors that are also strongly associated with such deaths. OBJECTIVES: The objective of this review was to assess the effects of antibiotics for preventing respiratory tract infections and overall mortality in adults receiving intensive care. SEARCH STRATEGY: We searched Medline, the Cochrane Acute Respiratory Infections Group trials register, proceedings of scientific meetings and reference lists of articles from January 1984 to December 1999. We also contacted investigators in the field. SELECTION CRITERIA: Randomised trials of antibiotic prophylaxis for respiratory tract infections and deaths among adult intensive care unit patients. DATA COLLECTION AND ANALYSIS: Investigators were contacted for additional information. At least two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Overall 33 trials involving 5727 people were included. There was variation in the antibiotics used, patient characteristics and risk of respiratory tract infections and mortality in the control groups. In 16 trials (involving 3361 patients) that tested a combination of topical and systemic antibiotic, the average rates of respiratory tract infections and deaths in the control group were 36% and 30% respectively. There was a significant reduction of both respiratory tract infections (odds ratio 0.35, 95% confidence interval 0.29 to 0.41) and total mortality (odds ratio 0.80, 95% confidence interval 0.69 to 0.93) in the treated group. On average 5 patients needed to be treated to prevent one infection and 23 patients to prevent one death. In 17 trials (involving 2366 patients) that tested topical antimicrobials the rates of respiratory tract infections and deaths in the control groups were 28% and 26% respectively. There was a significant reduction of respiratory tract infections (odds ratio 0.56, 95% confidence interval 0.46 to 0.68) but not in total mortality (odds ratio 1.01, 95% confidence interval 0.84 to 1.22) in the treated group. REVIEWER'S CONCLUSIONS: A combination of topical and systemic prophylactic antibiotics can reduce respiratory tract infections and overall mortality in adult patients receiving intensive care. The design of the trials included in this systematic review does not allow to assess whether or not the treatment leads to antimicrobial resistance. Trials with different design are warranted to reliably address this question. PMID- 11034668 TI - Antibiotics for sore throat. AB - BACKGROUND: Sore throat is a very common reason for people to attend for medical care. It is a disease that remits spontaneously, that is, 'cure' is not dependent on treatment. Nonetheless primary care doctors commonly prescribe antibiotics for sore throat and other upper respiratory tract infections. OBJECTIVES: To assess the benefits of antibiotics in the management of sore throat. SEARCH STRATEGY: Systematic search of the literature from 1945 to 1999, using electronic searches of MEDLINE (using the keywords, "pharyngitis", "sore throat" and "tonsillitis") after 1966, the Cochrane Library, the Cochrane collection of hand-searched trials, and the reference sections of the articles identified. Abstracts of identified articles were used to determine which studies were trials. SELECTION CRITERIA: Trials of antibiotic against control with either measures of the typical symptoms (throat soreness, headache or fever), or complications (suppurative and non-suppurative) of sore throat. DATA COLLECTION AND ANALYSIS: RevMan 4.0.3 MAIN RESULTS: 25 studies were included in the review. A total number of 11, 452 cases of sore throat have been studied. 1. Non-suppurative complications There was a trend for protection against acute glomerulonephritis by antibiotics, but insufficient cases were recorded to be sure of this effect. Several studies found benefit from antibiotics for acute rheumatic fever, which reduced this complication to less than one third (OR = 0.30; 95% CI = 0.20-0.45). 2. Suppurative complications Antibiotics reduced the incidence of acute otitis media to about one quarter of that in the placebo group (OR = 0.22; 95% CI = 0.11 0.43) and reduced the incidence of acute sinusitis to about one half of that in the placebo group (OR = 0.46; 95% CI = 0.10-2.05). The incidence of quinsy was also reduced in relation to placebo group (OR = 0.16; 95% CI = 0.07-0.35). 3. Symptoms Symptoms of headache, throat soreness and fever were reduced by antibiotics to about one half. The greatest time for this to be evident was at about three and a half days (when the symptoms of about 50% of untreated patients had settled). About 90% of treated and untreated patients were symptom-free by one week. 4. Subgroup analyses of symptom reduction Subgroup analysis by age; blind vs unblinded; or use of antipyretics yielded no significant differences. The results of swabs of the throat for Streptococcus influenced the effect of antibiotics. If the swab was positive, antibiotics were more effective (the OR reduced to 0.16, 95% CI 0.09, 0.26) than if it was negative (OR 0.65; 95% CI 0.38,1.1.2). REVIEWER'S CONCLUSIONS: Antibiotics confer relative benefits in the treatment of sore throat. However, the absolute benefits are modest. Protecting sore throat sufferers against suppurative and non-suppurative complications in modern Western society can only be achieved by treating many with antibiotics who will derive no benefit. Antibiotics shorten the duration of symptoms, but by a mean of only one day about half way through the illness (the time of maximal effect), and by about sixteen hours overall. PMID- 11034669 TI - Routine versus selective antifungal administration for control of fungal infections in patients with cancer. AB - BACKGROUND: Systemic fungal infection is considered to be an important cause of morbidity and mortality in cancer patients, particularly those with neutropenia. Antifungal drugs are often given prophylactically, or to patients with persistent fever. OBJECTIVES: The objective of this review was to assess the effect of antifungal drugs in cancer patients with neutropenia. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register and MEDLINE (November 1999) and the reference lists of articles. We searched the proceedings of the ICAAC, General Meeting of the ASM (from 1990 to 1999), and the 7th European Congress of Clinical Microbiology and Infectious Diseases (1995 to 1999) and contacted researchers in the field. SELECTION CRITERIA: Randomised trials of amphotericin B, AmBisome, fluconazole, ketoconazole, miconazole, or itraconazole compared with placebo or no treatment in cancer patients with neutropenia. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility, methodological quality and abstracted data. MAIN RESULTS: Twenty-nine trials involving 3875 patients were included. Intravenous amphotericin B reduced total mortality (relative risk 0.72, 95% confidence interval 0.51 to 1.02, P=0.06) based on 8 trials. This borderline result was confirmed by three trials which compared lipid soluble amphotericin B (AmBisome) with smaller doses of standard amphotericin B; the trials demonstrated an effect of AmBisome on mortality, relative risk 0.70 (95% CI 0.50 to 0.99). The risk difference for the two estimates combined is 0.040 (95% CI 0.012 to 0.068) which means that 25 patients (95% CI 15 to 83) would need to be treated with intravenous amphotericin B to avoid one death. In contrast, fluconazole, ketoconazole, miconazole and itraconazole had no effect on mortality. The incidence of invasive fungal infection decreased with administration of amphotericin B (relative risk 0.39, 95% CI 0.20 to 0.76), fluconazole (relative risk 0.39, 95% CI 0.27 to 0.57) and itraconazole (relative risk 0.45, 95% CI 0.20 to 0.99), but not with miconazole or ketoconazole. REVIEWER'S CONCLUSIONS: Intravenous amphotericin B is the only antifungal agent which has a documented effect on mortality, and there is not sufficient evidence to judge the relative merits of other antifungal agents. This drug should therefore be preferred for prophylactic or empirical antifungal therapy in cancer patients with neutropenia. PMID- 11034670 TI - Antidepressants for smoking cessation. AB - BACKGROUND: There are two reasons to believe antidepressants might help in smoking cessation. First, depression may be a symptom of nicotine withdrawal, and smoking cessation sometimes precipitates depression. Second, smoking appears to be due, in part, to deficits in dopamine, serotonin and norepinephrine, all of which are increased by antidepressants. OBJECTIVES: The aim of this review is to assess the effectiveness of antidepressant medications in aiding long term smoking cessation. The drugs include bupropion; doxepin; fluoxetine; imipramine; moclobemide; nortriptyline; selegiline; sertraline, tryptophan and venlafaxine. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register which includes trials indexed in Medline, Embase, SciSearch and PsycLit, and other reviews and meeting abstracts. SELECTION CRITERIA: We considered randomized trials comparing antidepressant drugs to placebo or an alternative therapeutic control for smoking cessation. We excluded trials with less than 6 months follow up. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of study population, the nature of the drug therapy, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least six months follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: There was one trial each of moclobemide, sertraline and venlafaxine, two of fluoxetine and nortriptyline, and five trials of bupropion, one of which tested long term use to prevent relapse. Nortriptyline and bupropion both increased cessation. In one trial the combination of bupropion and nicotine patch produced higher quit rates than patch alone. REVIEWER'S CONCLUSIONS: Some antidepressants (bupropion and nortriptyline) can aid smoking cessation. It is not clear whether these effects are specific for individual drugs, or a class effect. PMID- 11034671 TI - Combined inhaled anticholinergics and beta2-agonists for initial treatment of acute asthma in children. AB - BACKGROUND: Several randomized controlled trials have examined, with conflicting results, the efficacy of the addition of anticholinergics to beta2 agonists in acute pediatric asthma. The pooling for a larger number of randomized controlled trials may provide not only greater power for detecting group differences and also provide better insight into the influence of patients' characteristics and treatment modalities on efficacy. OBJECTIVES: The aims of this study were to estimate the therapeutic and adverse effects attributable to the addition of inhaled anticholinergics to beta2 agonists in acute pediatric asthma. SEARCH STRATEGY: We searched Medline (1966 to April 2000), Embase (1980 to April 2000), Cinahl (1982 to April 2000) and reference lists of studies. We also contacted drug manufacturers and trialists. SELECTION CRITERIA: Randomised trials comparing the combination of inhaled anticholinergics and beta2 agonists with beta2 agonists alone in children aged 18 months to 17 years with acute asthma. DATA COLLECTION AND ANALYSIS: Assessments of trial quality and data extraction were done by two reviewers independently. MAIN RESULTS: Of the 40 identified trials, 13 were relevant and eight of these were of high quality. The addition of a single dose of anticholinergic to beta2 agonists did not reduce hospital admission [RR=0.93 (95% CI: 0.65, 1.32)]. However, significant group differences in lung function supporting the combination of anticholinergics and beta2 agonists were observed 60 minutes [SMD=0.57 (95% CI:0.21, 0.93)] and 120 minutes [SMD=0.53 (95% CI: 0.17, 0.90)] after the dose of anticholinergic. In contrast, the addition of multiple doses of anticholinergics to beta2 agonists reduced the risk of hospital admission by 25% [RR=0.75 (95% CI: 0.62,0.89)] in children with predominantly moderate and severe exacerbations. Twelve (95% CI: 8, 32) children would need to be treated to avoid one admission. When restricting this strategy to children with severe exacerbations, seven (95% CI: 5, 20) children need to be treated to avoid an admission. At 60 minutes after the last anticholinergic inhalation, a weighted mean group difference of 9.68 in change in % predicted FEV1 [95% CI:5.70, 13.68] favored anticholinergic use. In the two studies where anticholinergics were systematically added to every beta2 agonist inhalation, irrespective of asthma severity, no group differences were observed for the few available outcomes. There was no increase in the amount of nausea, vomiting or tremor in patients treated with anticholinergics. REVIEWER'S CONCLUSIONS: A single dose of an anticholinergic agent is not effective for the treatment of mild and moderate exacerbations and is insufficient for the treatment of severe exacerbations. Adding multiple doses of anticholinergics to beta2 agonists appears safe, improves lung function and would avoid hospital admission in 1 of 12 such treated patients. Although multiple doses should be preferred to single doses of anticholinergics, the available evidence only supports their use in school-aged children with severe asthma exacerbation. There is no conclusive evidence for using multiple doses of anticholinergics in children with mild or moderate exacerbations. PMID- 11034672 TI - Doxapram versus methylxanthine for apnea in preterm infants. AB - BACKGROUND: Recurrent apnea is common in preterm infants, particularly at very early gestational ages. These episodes of loss of effective breathing can lead to hypoxemia and bradycardia which may be severe enough to require resuscitation including use of positive pressure ventilation. Doxapram and methylxanthine drugs have been used to stimulate breathing and so prevent apnea and its consequences. OBJECTIVES: To assess the effects of doxapram compared with methylxanthine in preterm infants with recurrent apnea. SEARCH STRATEGY: The Cochrane Collaboration Clinical Trials Register (Cochrane Library issue 3, 2000), MEDLINE (1966- July 2000), reference lists of relevant articles and conference proceedings. SELECTION CRITERIA: Randomized and quasi-randomized trials of doxapram compared with methylxanthine (e.g. theophylline, aminophylline or caffeine) for the treatment of apnea in preterm infants. DATA COLLECTION AND ANALYSIS: The methodological quality of each trial was reviewed by the second reviewer blinded to trial authors and institution(s). Additional information was requested from authors. Each reviewer extracted the data separately, then they were compared and differences resolved. Meta-analysis was carried out with use of relative risk and risk difference. MAIN RESULTS: Three trials involving 56 infants were included. No difference was detected between intravenous doxapram or methylxanthine in the incidence of failed treatment within 48 hours (relative risk 1.16, 95% confidence interval 0.43 to 3.13). No infants were reported to have been given mechanical ventilation on either treatment. No adverse effects were reported. REVIEWER'S CONCLUSIONS: Intravenous doxapram and intravenous methylxanthine appear to be similar in their short term effects for treating apnea in preterm infants, although these trials are too small to exclude an important difference between the two treatments or to exclude the possibility of less common adverse effects. Longer term outcome of infants treated in these trials has not been reported. Further studies would require a large number of infants to clarify whether there might be differences in responses or adverse effects with these two drugs at different ages. PMID- 11034673 TI - Glycerol for acute stroke. AB - BACKGROUND: Brain oedema is a major cause of early death after stroke. A 10% solution of glycerol is a hyperosmolar agent that is claimed to reduce brain oedema. OBJECTIVES: To determine whether I.V. glycerol treatment in acute stroke, either ischaemic or haemorrhagic, influences death rates and functional outcome in the short or long term and whether the treatment is safe. SEARCH STRATEGY: The Cochrane Stroke Group Trials Register was searched, conference proceedings were screened and some trialists were personally contacted. SELECTION CRITERIA: All completed, randomised and quasi-randomized, controlled, published and unpublished comparisons, evaluating clinical outcome in which intravenous (I.V.) glycerol treatment was initiated within the first days after stroke onset. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied the inclusion criteria, assessed the trial quality and extracted data and this was checked with all co reviewers. Death from all causes, functional outcome and adverse effects were analysed. MAIN RESULTS: Eleven completed, randomised trials comparing I.V. glycerol and control were considered. Analysis of death during the scheduled treatment period for acute ischaemic and/or haemorrhagic stroke was possible in ten trials where 482 glycerol treated patients were compared with 463 control patients. Glycerol was associated with a non-significant reduction in the odds of death within the scheduled treatment period (OR 0.78, 95% Confidence Intervals 0.58 - 1.06). Among patients with definite or probable ischaemic stroke, glycerol was associated with a significant reduction in the odds of death during the scheduled treatment period (odds ratio 0.65, 95% CI 0.44-0.97). However, at the end of the scheduled follow up period, there was no significant difference in the odds of death (odds ratio 0.98, 95% CI 0.73-1.31). Functional outcome was reported in only two studies but there were non-significantly more patients who had a good outcome at the end of scheduled follow up (odds ratio 0.73, 95% CI 0.37-1.42). Haemolysis seems to be the only relevant adverse effect of glycerol treatment. REVIEWER'S CONCLUSIONS: This systematic review suggests a favourable effect of glycerol treatment on short term survival in patients with probable or definite ischaemic stroke but the magnitude of the treatment effect may be minimal (as low as a 3% reduction in odds). Due to the relatively small number of patients and that the trials have been performed in the pre-CT era, the results must be interpreted cautiously. The lack of evidence of benefit in long term survival does not support the routine or selective use of glycerol treatment in patients with acute stroke. PMID- 11034674 TI - Co-ordinated multidisciplinary approaches for inpatient rehabilitation of older patients with proximal femoral fractures. AB - BACKGROUND: Hip fracture is a major cause of morbidity in older people and its impact, both on the individual and to society, is substantial. OBJECTIVES: To examine the effects of co-ordinated multidisciplinary inpatient rehabilitation, compared with usual orthopaedic care, for older patients with hip fracture. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, Medline (up to August 1999), and reference lists of published papers and books. We also contacted colleagues and trialists. SELECTION CRITERIA: Randomised and quasi-randomised trials of post surgical care using specialised rehabilitation of mainly older patients (aged 65 years or over) with hip fracture. DATA COLLECTION AND ANALYSIS: Trial assignment to included, excluded and awaiting assessment categories, was by consensus. Two reviewers independently assessed trial quality and extracted data. Limited additional information was sought from most trialists. As well as pooling of data from primary outcomes, supplementary analyses were performed to combine clinically relevant outcomes and investigate possible explanatory factors. MAIN RESULTS: In this substantive update, three new trials have been included. The eight included trials involved 1609 patients. The combined outcomes of death or requiring institutional care showed no significant difference between intervention and control groups (relative risk 0.91; 95% confidence interval 0.80 to 1.03). There was considerable heterogeneity in length of stay and cost data. Using death and deterioration in function as a further combined outcome variable yielded a relative risk of 0.90 (95% confidence interval 0.81 to 1.01). This should be interpreted with caution due to heterogeneity. No quality of life measures were reported and the two trials investigating carer burden showed no detrimental effect from the intervention. The review update did not result in any new data for these outcomes. REVIEWER'S CONCLUSIONS: The trials reviewed had different aims, interventions and outcomes. As a consequence, results were heterogeneous and the question of effectiveness of different types of co-ordinated inpatient rehabilitation after hip fracture cannot be answered conclusively. There is a trend to effectiveness when combined outcome variables (death and institutional care, death and deterioration in function) are considered. Future trials of post surgical care involving inpatient rehabilitation, or other models such as 'early supported discharge' and 'hospital at home' schemes, should aim to establish both effectiveness and cost effectiveness of multidisciplinary rehabilitation overall, rather than attempt to evaluate its components. PMID- 11034675 TI - Mefloquine for preventing malaria in non-immune adult travellers. AB - BACKGROUND: Mefloquine is commonly prescribed to prevent malaria in travellers, and has replaced other drugs because Plasmodium falciparum is commonly resistant to them. However, mefloquine may be associated with neuropsychiatric harmful effects. OBJECTIVES: To assess the effects of mefloquine in adult travellers compared to other regimens in relation to episodes of malaria, withdrawal from prophylaxis, and adverse events. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, MEDLINE, EMBASE, LILACS, Science Citation Index and bibliographies in retrieved papers and standard textbooks. We contacted researchers in the subject of malaria chemoprophylaxis, and drug companies. SELECTION CRITERIA: Randomised trials comparing mefloquine with other standard prophylaxis or placebo in non-immune adult travellers, and in non travelling volunteers. For adverse events, any published case reports were collected. DATA COLLECTION AND ANALYSIS: We independently assessed trial quality and extracted data. Adverse events from observational studies were categorised by the study type. We also contacted study authors. MAIN RESULTS: We included 10 trials involving 2750 non-immune adult participants. Five of these were field trials, and of these all were in mainly male soldiers. One trial comparing mefloquine with placebo showed mefloquine prevented malaria episodes in an area of drug resistance (odds ratio 0.04, 95% confidence interval 0.02 to 0.08). Withdrawals in the mefloquine group were consistently higher in four placebo controlled trials (odds ratio 3.56, 95% confidence interval 1.67 to 7.60). In five trials comparing mefloquine with other chemoprophylaxis, no difference in tolerability was detected. We found 516 published case reports of mefloquine adverse effects. 63 per cent of these published reports involved tourists and business travellers. There were four fatalities attributed to mefloquine. REVIEWER'S CONCLUSIONS: Mefloquine prevents malaria, but has adverse effects that limit its acceptability. There is evidence from non-randomised studies that mefloquine has potentially harmful effects in tourists and business travellers, and its use needs to be carefully balanced against this. Trials of comparative effects of antimalarial prophylaxis should include episodes of malaria and withdrawal from prophylaxis as outcomes. PMID- 11034676 TI - Drugs for preventing tuberculosis in HIV infected persons. AB - BACKGROUND: People with HIV have a increased risk of developing tuberculosis. Preventive therapy may help prevent progression of tuberculosis infection to disease. OBJECTIVES: The objective of this review was to assess the effects of preventive therapy with anti-tuberculosis drugs in people with HIV infection. SEARCH STRATEGY: The Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, Embase and reference lists of articles were searched. Researchers in the field were contacted. SELECTION CRITERIA: Randomised trials of anti-tuberculosis drugs in people with HIV infection but without evidence of active tuberculosis. DATA COLLECTION AND ANALYSIS: One reviewer assessed eligibility and trial quality. Study authors were contacted for additional information. MAIN RESULTS: Seven trials were included. Compared to placebo, preventive therapy was associated with a lower incidence of active tuberculosis (Peto odds ratio 0.54, 95% confidence interval 0.39 to 0.76). Risk of death (OR 0.96 95%CI 0.82 to 1.13) was not significantly different in the two groups. Incidence of tuberculosis was reduced in people with a positive tuberculin skin test (OR 0.35, 95% confidence interval 0.21 to 0.59), but was not significantly lower in those with a negative skin test (OR 0.82, 95% confidence interval 0.51 to 1.31). Similarly death was less frequent in those with a positive skin test who received preventive therapy (OR 0.70, 95% confidence interval 0.50 to 0/98), but this difference was not observed among those with a negative skin test OR 1.04, 95% confidence interval 0.86 to 1.28). Each regimen (isoniazid alone, isoniazid plus rifampicin, isoniazid plus rifampicin plus pyrazinamide, rifampicin plus pyrazinamide) had similar protective effects against active tuberculosis. REVIEWER'S CONCLUSIONS: Preventive therapy appears to be effective in reducing incidence of tuberculosis, and death from tuberculosis in HIV infected adults with a positive tuberculin skin test, at least in the short to medium term. Choice of regimen will depend on issues of adherence, side effects profile, cost and drug resistance. PMID- 11034677 TI - Antibiotics for acute otitis media in children. AB - BACKGROUND: Acute otitis media is one of the most common diseases in early infancy and childhood. Antibiotic use for acute otitis media varies from 31% in the Netherlands to 98% in the USA and Australia. OBJECTIVES: The objective of this review was to assess the effects of antibiotics for children with acute otitis media. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE, Index Medicus (pre 1965), Current Contents and reference lists of articles from 1958 to January 2000. SELECTION CRITERIA: Randomised trials comparing antimicrobial drugs with placebo in children with acute otitis media. DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Ten trials were eligible but only seven trials, with a total of 2,202 children, included patient-relevant outcomes. The methodological quality of the included trials was generally high. All trials were from developed countries. The trials showed no reduction in pain at 24 hours, but a 28% relative reduction (95% confidence interval 15% to 38%) in pain at two to seven days. Since approximately 80% of patients will have settled spontaneously in this time, this means an absolute reduction of 5% or that about 17 children must be treated with antibiotics to prevent one child having some pain after two days. There was no effect of antibiotics on hearing problems of acute otitis media, as measured by subsequent tympanometry. However, audiometry was done in only two studies and incompletely reported. Nor did antibiotics influence other complications or recurrence. There were few serious complications seen in these trials: only one case of mastoiditis occurred in a penicillin treated group. REVIEWER'S CONCLUSIONS: Antibiotics provide a small benefit for acute otitis media in children. As most cases will resolve spontaneously, this benefit must be weighed against the possible adverse reactions. Antibiotic treatment may play an important role in reducing the risk of mastoiditis in populations where it is more common. [This abstract has been prepared centrally.] PMID- 11034678 TI - Antibiotics for acute bronchitis. AB - BACKGROUND: Antibiotic treatment of acute bronchitis, which is one of the most common illnesses seen in primary care, is controversial. Most clinicians prescribe antibiotics in spite of expert recommendations against this practice. OBJECTIVES: The objective of this review was to assess the effects of antibiotic treatment for patients with a clinical diagnosis of acute bronchitis. SEARCH STRATEGY: We searched MEDLINE, EMBASE, reference lists of articles and the authors' personal collections up to 1996, and Scisearch from 1989 to 1996; we also wrote to study authors and drug manufacturers. An updated search of the Cochrane Controlled Trials Register and MEDLINE was conducted in 2000. SELECTION CRITERIA: Randomised trials comparing any antibiotic therapy with placebo in acute bronchitis or acute productive cough without other obvious cause in patients without underlying pulmonary disease. DATA COLLECTION AND ANALYSIS: At least two reviewers extracted data and assessed trial quality. Authors were contacted for missing data. MAIN RESULTS: Nine trials involving over 750 patients aged eight to over 65 and including smokers and non-smokers were included. The quality of the trials was variable. A variety of outcome measures were assessed. Overall, patients receiving antibiotics had better outcomes than did those receiving placebo. At a follow-up visit, they were less likely to have a cough (relative risk 0.64, 95% confidence interval 0.49 to 0.85; number needed to treat 5, 95% CI 3 to 14), show no improvement on physician assessment (RR 0.52, 95% CI 0.31 to 0.87; NNT 14, 95% CI 8 to 50), or have abnormal lung findings (RR 0.48, 95% CI 0.26 to 0.89; NNT 11, 95% CI 6 to 50); and had shorter durations of cough (weighted mean difference 0.58 days, 95% CI 0.01 to 1.16 days), productive cough (WMD 0.52 days, 95% CI 0.01 to 1.03 days), and feeling ill (WMD 0.58 days, 95% CI 0.00 to 1.16 days). There were no significant differences regarding the presence of night cough, productive cough, or activity limitations at follow-up, or in the mean duration of activity limitations. The benefits of antibiotics were less apparent in a sensitivity analysis that included data from two other studies of patients with upper respiratory tract infections with productive cough. Antibiotic-treated patients reported significantly more adverse effects (RR 1.48, 95% CI 1.02 to 2.14; number needed to harm 17, 95% CI 9 to 100) such as nausea, vomiting, headache, skin rash or vaginitis. REVIEWER'S CONCLUSIONS: Overall, antibiotics appear to have a modest beneficial effect in patients who are diagnosed with acute bronchitis. The magnitude of this benefit, however, is similar to that of the detriment from potential adverse effects. Furthermore, patients with other symptoms of the common cold who have been ill for less than one week are not likely to have any benefit from antibiotics. PMID- 11034679 TI - Antifibrinolytics for heavy menstrual bleeding. AB - BACKGROUND: Heavy menstrual bleeding (HMB) is an important cause of ill health in women. Medical therapy, with the avoidance of possibly unnecessary surgery, is an attractive treatment option. A wide variety of medications are available to reduce heavy menstrual bleeding but there is considerable variation in practice and uncertainty about the most appropriate therapy. Plasminogen activators are a group of enzymes that cause fibrinolysis (the dissolution of clots). An increase in the levels of plasminogen activators has been found in the endometrium of women with heavy menstrual bleeding compared to those with normal menstrual loss. Plasminogen activator inhibitors (antifibrinolytic agents) have therefore been promoted as a treatment for heavy menstrual bleeding. There has been a reluctance to prescribe tranexamic acid due to possible side effects of the drugs such as an increased risk of thrombogenic disease (deep venous thrombosis). Long term studies in Sweden, however, have shown that the rate of incidence of thrombosis in women treated with tranexamic acid is comparable with the spontaneous frequency of thrombosis in women. OBJECTIVES: To determine the effectiveness of antifibrinolytics in achieving a reduction in heavy menstrual bleeding. SEARCH STRATEGY: All studies which might describe randomised controlled trials of antifibrinolytic therapy for the treatment of heavy menstrual bleeding were obtained by electronic searches of MEDLINE 1966-1997, EMBASE 1980-1997 and the Cochrane Library. Companies producing antifibrinolytics and experts within the field were contacted for reference lists and information on unpublished trials. SELECTION CRITERIA: Randomised controlled trials in women of reproductive age treated with antifibrinolytic agents versus placebo, no treatment or any other medical (non-surgical) therapy for regular heavy menstrual bleeding within either the primary, family planning or specialist clinic settings. Women with post menopausal bleeding, intermenstrual bleeding, iatrogenic or pathological causes of heavy menstrual bleeding were excluded. DATA COLLECTION AND ANALYSIS: Fifteen eligible trials were assessed by three reviewers and eight of these did not meet with the inclusion criteria. Of the seven remaining trials, four of these could be included within the meta-analysis. The remaining three trials had a crossover design and despite contacting the authors and appropriate companies, we were unable to extract the results in a format suitable to include these within the meta-analysis. However the results are included within the text of the review for discussion. MAIN RESULTS: Antifibrinolytic therapy compared to placebo showed a significant reduction in mean blood loss (WMD -94.0 [-151.4, -36.5]) and significant change in mean reduction of blood loss (WMD -110.2 [-146.5, -73.8]). This objective improvement was not mirrored by a patient perceived improvement in monthly menstrual blood loss (RR 2.5 [0.9, 7.3]) in the one study which recorded this outcome (~~ Edlund 1995~~). Antifibrinolytic agents were compared to only three other medical (non-surgical) therapies: mefenamic acid, norethisterone administered in the luteal phase and ethamsylate. In all instances, there was a significant reduction in mean blood loss (WMD -73.0 [-123.4, -22.6], WMD -111.0 [ 178.5, -43.5] and (WMD -100 [-143.9, -56.1] respectively) and a strong, although non-significant trend in favour of tranexamic acid in the participants' perception of an improvement in menstrual blood loss. There were no significant differences in the frequency of reported side effects with tranexamic acid when compared to oral luteal phase progestogens (RR 0.4 [0.1, 1.2]) or withdrawal from treatment because of adverse events when compared with NSAIDs and ethamsylate when these treatments were used for heavy menstrual bleeding. Change in the quality of life measures, flooding and leakage and sex life, were significantly improved in the tranexamic acid group when compared to the oral progestagen group. These findings are based in most cases on only one trial. REVIEWER'S CONCLUSIONS: Antifibrinolytic therapy causes a greater reduction in objective measurements of heavy menstrual bleeding when compared to placebo or other medical therapies (NSAIDS, oral luteal phase progestagens and ethamsylate). This treatment is not associated with an increase in side effects compared to placebo, NSAIDS, oral luteal phase progestagens or ethamsylate. Flooding and leakage and sex life is significantly improved after tranexamic acid therapy when compared with oral luteal progestogens but no other measures of quality of life were assessed. No study has used resource cost as an outcome. There are no data available within randomised controlled trials which record the frequency of thromboembolic events. PMID- 11034680 TI - Antithrombotic drugs for carotid artery dissection. AB - BACKGROUND: Extracranial internal carotid artery dissection can lead to occlusion of the artery and hence cause an ischemic stroke. It is the underlying stroke mechanism in approximately 2.5% of all strokes, and the second leading cause of stroke in patients younger than 45 years of age. Antithrombotic agents (heparin, oral anticoagulants or antiplatelet drugs) may prevent arterial thrombosis in eICAD, but these benefits may be offset by increased bleeding. OBJECTIVES: To determine whether antithrombotic drugs (antiplatelet drugs, anticoagulation) are effective and safe in treatment of patients with extracranial internal carotid artery dissection, and which is the better treatment. SEARCH STRATEGY: We searched the Cochrane Stroke Group specialised Trials Register for relevant randomised trials and controlled clinical trials. In addition, we performed comprehensive searches of MEDLINE and EMBASE and checked reference lists of all relevant papers for additional eligible studies. SELECTION CRITERIA: Randomised controlled trials, controlled clinical trials assessing the efficacy of antiplatelet drugs or anticoagulants in the treatment of extracranial carotid artery dissection. For non-randomised trials, case series (studies), that reported on antithrombotic treatment with at least 4 patients, were eligible. All trials and studies were assessed for eligibility. Data from all eligible studies were extracted independently by two reviewers. Disagreements were resolved by discussion. DATA COLLECTION AND ANALYSIS: Data on the primary outcome measures were extracted systematically. These were: all deaths, vascular deaths, and disability. Secondary outcomes were: first stroke occurrence, stroke recurrence, any stroke during reported follow-up, extracranial haemorrhage, and intracranial haemorrhage. The first choice treatment was taken for analyses. MAIN RESULTS: No randomised trials and 49 case series (including 683 treated patients) were identified. No reliable comparisons of antiplatelet drugs or anticoagulants with control were available. 24 eligible studies including 286 patients (who either received antiplatelet drugs or anticoagulants) were included in the analysis. There was no significant difference in odds of death comparing antiplatelet drugs with anticoagulants, odds ratio (OR) 2.41, 95% CI 0.27-21.80). There was also no significant difference in the odds of being alive but disabled (OR 1.65, 95% CI 0.50-5.42). Few major haemorrhages were reported. REVIEWER'S CONCLUSIONS: There were no randomised trials comparing either anticoagulants or antiplatelet drugs with control. There is, therefore, no evidence to support their routine use for the treatment of extracranial internal carotid artery dissection. There were also no randomised trials, that directly compared anticoagulants with antiplatelet drugs, and the reported non-randomised studies did not show any evidence of a significant difference between the two. We suggest that a randomised trial including at least 1000 patients in each treatment arm with this condition is clearly needed. PMID- 11034681 TI - Cytidinediphosphocholine (CDP choline) for cognitive and behavioural disturbances associated with chronic cerebral disorders in the elderly. AB - BACKGROUND: CDP-choline has a widespread, but not exclusive use in the treatment of disorders of a cerebrovascular nature. The many years of its use have caused an evolution in dosage, method of administration, and selection of patients to which the treatment was given. Design of the clinical studies, including length of observation, severity of disease, and methodology of evaluation of the results have also varied. In spite of uncertainties about its efficacy, CDP-choline is frequently prescribed for cognitive impairment in several continental European countries, especially when the clinical picture is predominantly one of cerebrovascular disease. OBJECTIVES: The objective is to assess the efficacy of CDP-choline (cytidinediphosphocholine) in the treatment of cognitive, emotional, and behavioural deficits associated with chronic cerebral disorders of older people. SEARCH STRATEGY: The CDCIG register of trials and other databases were searched in July 2000 for all relevant, non-animal randomized controlled trials using the terms CDP-choline/CDP, Citicoline, Cytidine Diphosphate choline and Diphosphocholine. The Psychlit (1974-1996), Psychiatry (1980-1996) and MEDLINE electronic databases have been searched independently by the reviewers. The reviewers have also contacted manufacturers of CDP-choline. SELECTION CRITERIA: All relevant, non-animal, unconfounded, double-blind, placebo-controlled, randomized trials of CDP-choline for patients with cognitive impairment due to chronic cerebral disorders are considered for inclusion in the review. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed the included studies, extracted the data, and pooled when appropriate and possible. The pooled odd ratios (95% CI) or the average differences (95% CI) were estimated. No intention-to-treat data were available from the studies included. MAIN RESULTS: Seven of the included studies observed the subjects for a period between 20 to 30 days, one study was of 6 weeks duration, 3 studies used cycles extending over 2 and 3 months and one study observed continuous administration over 3 months. The studies differed in dose, inclusion criteria for subjects, and outcome measures. Results are reported for the domains of attention, memory testing, behavioural rating scales, global clinical impression and tolerability. There is no significant evidence of a beneficial effect of CDP-choline on attention. There are modest, but statistically significant, beneficial effects of CDP-choline on memory function and behaviour. For the outcome of clinical global impression, the odds ratio for improvement in the subjects treated with CDP-choline as opposed to the subjects treated with placebo is 8.89 [5.19 to 15.22]. The drug is well tolerated. REVIEWER'S CONCLUSIONS: There is some evidence that CDP-choline has a positive effect on memory and behaviour in at least the short term. The evidence of benefit from global impression is stronger, but is still limited by the duration of the studies. There is evidence that the effect of treatment is more homogeneous for patients with cognitive impairment secondary to cerebrovascular disorder. Further studies with a more appropriate length of treatment are recommended owing to the chronic and irreversible nature of the disorders for which this treatment is indicated. PMID- 11034682 TI - Discharge planning from hospital to home. AB - BACKGROUND: Discharge planning is the development of an individualised discharge plan for the patient prior to leaving hospital for home, with the aim of containing costs and improving patient outcomes. It has been suggested that discharge planning can reduce unplanned readmission to hospital. OBJECTIVES: To assess the effects of discharge planning for patients moving from hospital to home. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register, the Cochrane Controlled Trials Register, MEDLINE (1966 to 1996), EMBASE (1980 to 1996), Sigle (1980 to 1996), Bioethics (1985 to 1996), Health Plan (all available years), PsycLit (1974 to 1996), Cinahl (1982 to 1996), EconLit (1969 to 1996), Social Science Citation Index (1992 to 1996), and reference lists of articles. SELECTION CRITERIA: Randomised trials and controlled trials comparing discharge planning with routine discharge for hospital patients. The outcomes were mortality, clinical complications, hospital length of stay, readmissions, discharge destination, general and disease specific health status, functional status, psychological well being, patient satisfaction, carer satisfaction, carer burden, cost to the health service, patient and family, general practice, and community services. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Eight studies were included involving 4837 patients. Four studies recruited patients with a medical condition; four recruited patients with a mix of medical and surgical conditions, and one of these recruited medical and surgical patients as separate groups. There was a small reduction in hospital length of stay for elderly medical patients allocated to discharge planning (weighted mean difference -1.01, 95% CI -2.06 to 0.05). The effects of discharge planning on readmission rates were mixed. No statistically significant differences were detected for patient health outcomes. Patients with medical conditions allocated to discharge planning reported increased satisfaction compared with those receiving routine discharge. No statistically significant differences were reported for overall health care costs, although one study reported a significant reduction in readmission costs for medical patients allocated to discharge planning. REVIEWER'S CONCLUSIONS: The studies showed mixed results, which may reflect the different study populations and the different ways the intervention was implemented. There is some evidence that discharge planning may lead to reduced hospital length of stay, and in some cases reduced readmission to hospital. There is also some evidence that discharge planning increased patient satisfaction. There was no evidence that discharge planning reduced health care costs; however few studies conducted a formal economic analysis. PMID- 11034683 TI - Conservative versus operative treatment for hip fractures. AB - BACKGROUND: Until operative treatment involving the use of various implants was introduced in the 1950s, hip fractures were managed using conservative methods based on traction and bed rest. OBJECTIVES: To compare conservative with operative treatment for fractures of the proximal femur (hip) in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register and bibliographies of published papers, and contacted trialists. Date of the most recent search: August 1999. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing these two treatment methods in adults with hip fracture. Outcomes sought fell into four categories: a) fracture fixation complications, b) post-operative or clinical complications, c) anatomical restoration and d) final outcome measures including mortality. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, by use of an twelve item scale, and extracted data. Additional information was sought from trialists. After grouping by fracture type, comparable groups of trials were subgrouped by implant type and where appropriate, data were pooled using the fixed effects model. MAIN RESULTS: The five randomised trials identified involved only 425 elderly patients. One small and potentially biased trial of 23 patients with undisplaced intracapsular fracture showed a reduced risk of non-union for those fractures treated operatively. The four identified studies on extracapsular fractures tested a variety of surgical techniques and implant devices and only one trial involving 106 patients can be considered to test current practice. In this trial, no differences were found in medical complications, mortality and long-term pain. However, operative treatment was more likely to result in the fracture healing without leg shortening, a shorter hospital stay and a statistically non significant increase in the return of patients back to their original residence. REVIEWER'S CONCLUSIONS: Given the lack of available evidence to inform practice and the continued variation in practice, good quality randomised trials of operative versus conservative treatment for undisplaced intracapsular fractures are warranted. The limited available evidence from randomised trials does not suggest major differences in outcome between conservative and operative management programmes for extracapsular femoral fractures, but operative treatment appears to be associated with a reduced length of hospital stay and improved rehabilitation. However these results are derived mainly from one study. Conservative treatment will be acceptable where modern surgical facilities are unavailable, and will result in a reduction in complications associated with surgery, but rehabilitation is likely to be slower and limb deformity more common. Although further randomised trials would provide more robust data, they may be difficult to mount. PMID- 11034684 TI - Vaccines for preventing influenza in people with asthma (Cochrane Review). AB - BACKGROUND: Influenza vaccination is recommended for asthmatic patients in many countries as observational studies have shown that influenza infection can be associated with asthma exacerbations, but influenza vaccination itself has the potential to adversely affect pulmonary function. A recent overview concluded that there was no clear benefit of influenza vaccination in patients with asthma but this conclusions was not based on a systematic search of the literature. OBJECTIVES: Whilst influenza may cause asthma exacerbations, there is controversy about the use of influenza vaccinations, since they may precipitate an asthma attack in some people. The objective of this review was to assess the effects of influenza vaccination in children and adults with asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and checked reference lists of articles. SELECTION CRITERIA: Randomised trials of influenza vaccination in children (over two years of age) and adults with asthma. Studies involving people with chronic obstructive pulmonary disease were excluded. DATA COLLECTION AND ANALYSIS: Inclusion criteria and assessment of trial quality were applied by two reviewers independently. Data extraction was done by two reviewers independently. Study authors were contacted for missing information. MAIN RESULTS: Nine trials were included. Four of these trials were of high quality. One further article has been included since the previous version of this review. Inclusion of the new trial has not altered the conclusions of this review. The included studies covered a wide diversity of people, settings and types of influenza vaccination, so data from the different trials were not pooled. In one trial, no protective effect of influenza vaccination against asthma exacerbation was demonstrated, but the incidence of influenza was low during the study period. A higher number of asthma exacerbations following killed influenza vaccination was found in one trial (risk difference 3 1%, 95% confidence interval 0.3% to 5.8%). When people with upper respiratory tract infections were excluded, this difference was no longer significant. A small trial using recombinant vaccine found no significant difference in asthma exacerbations between the vaccinated and placebo groups. Updated search conducted July, 2000. No new trials were identified. REVIEWER'S CONCLUSIONS: There is not enough evidence to assess the benefits and risks of influenza vaccination for people with asthma. PMID- 11034685 TI - Inositol for respiratory distress syndrome in preterm infants. AB - BACKGROUND: Inositol is an essential nutrient required by human cells in culture for growth and survival. Inositol promotes maturation of several components of surfactant and may play a critical role in fetal and early neonatal life. OBJECTIVES: To assess the effectiveness/safety of supplementary inositol in preterm infants with RDS in reducing adverse neonatal outcomes. SEARCH STRATEGY: Medline, Embase, and Reference Update Databases were searched in August 1997 using key words: inositol and infant-newborn and random allocation or controlled trial or randomized trial (RCT). The reference lists of identified RCTs, personal files and Science Citation Index were searched. Unpublished additional information was obtained from the authors of one RCT published in abstract form. SELECTION CRITERIA: All randomized controlled trials of inositol supplementation to preterm infants with a control group that received a placebo or no intervention were included. Outcomes of interest were bronchopulmonary dysplasia (BPD), death, BPD or death, retinopathy of prematurity (ROP), intraventricular hemorrhage (IVH), necrotizing enterocolitis (NEC), and sepsis. DATA COLLECTION AND ANALYSIS: Data on neonatal outcomes were abstracted independently by the two researchers and any discrepancy was resolved through consensus. Revman was used for analysis of the data. MAIN RESULTS: Four reports of three RCTs were identified. One report was a duplicate publication. The outcome of death or bronchopulmonary dysplasia was reported in two trials, and was found to be significantly reduced (RR 0.56, 95% CI 0.42, 0.77; RD -0.215, 95% CI -0.323, 0.107). The outcome of death was reported in two trials and was found to be significantly reduced (RR 0.48, 95% CI 0.28, 0.80; RD -0.131, 95% CI -0.218, 0.043). Retinopathy of prematurity, stage 4 or needing therapy, was reported in two trials, and was found to be significantly reduced (RR 0.09, 95% CI 0.01, 0.67; RD -0.078, 95% CI -0.128, -0.027). Intraventricular hemorrhage, grade III IV, was significantly decreased (RR 0.55, 95% CI 0.32, 0.95; RD -0.090, 95% CI 0.170, -0.010). Neither sepsis nor necrotizing enterocolitis outcomes were increased. When a secondary analysis was done excluding a study published in abstract form, the results differed only in that there was a significant reduction in retinopathy of prematurity, any stage (RR 0.53, 95% CI 0.29, 0.97; RD -0.082, 95% CI -0.159,-0.005). REVIEWER'S CONCLUSIONS: Inositol supplementation results in statistically significant and clinically important reductions in important short-term adverse neonatal outcomes. A multi-center RCT of appropriate size is justified to confirm these findings. PMID- 11034686 TI - Interventions for treating chronic pelvic pain in women. AB - BACKGROUND: Chronic pelvic pain is common in women in the reproductive age group and it causes disability and distress and results in significant costs to health services. The pathogenesis of chronic pelvic pain is poorly understood. Often, investigation by laparoscopy reveals no obvious cause for pain. There are several possible explanations for chronic pelvic pain including undetected irritable bowel syndrome, the vascular hypothesis where pain is thought to arise from dilated pelvic veins in which blood flow is markedly reduced and altered spinal cord and brain processing of stimuli in women with chronic pelvic pain. As the pathophysiology of chronic pelvic pain is not well understood, its treatment is often unsatisfactory and limited to symptom relief. Currently, the main approaches to treatment include counseling or psychotherapy, attempting to provide reassurance using laparoscopy to exclude serious pathology, progestogen therapy such as medroxyprogesterone acetate, and surgery to interrupt nerve pathways. OBJECTIVES: We aimed to identify and review treatments for chronic pelvic pain in women in the reproductive years. The review included studies of patients with a diagnosis of pelvic congestion syndrome or adhesions but excluded those with pain known to be caused by i) endometriosis, ii) primary dysmenorrhoea (period pain), iii) pain due to active chronic pelvic inflammatory disease, or iv) irritable bowel syndrome. SEARCH STRATEGY: The search strategy adopted by the Cochrane Menstrual Disorders and Subfertility Group was used. SELECTION CRITERIA: Randomised controlled trials (RCTs) with women who had chronic pelvic pain, excluding endometriosis, primary dysmenorrhoea, pain due to chronic pelvic inflammatory disease, or irritable bowel syndrome. The reviewers were prepared to consider studies of any intervention including lifestyle, physical, medical, surgical and psychological treatments. Outcome measures were pain rating scales, quality of life measures, economic analyses and adverse events. DATA COLLECTION AND ANALYSIS: For each included trial, information was collected regarding the method of randomisation, allocation concealment, blinding, whether an intention to treat analysis could possibly be performed and relevant interventions and outcomes (see previous sections). Data were extracted independently by the two reviewers, using forms designed according to the Cochrane guidelines. MAIN RESULTS: Nine studies were identified of which five were of good methodological quality. Two studies were reported in a brief abstract only and were excluded. Progestogen (Medroxyprogesterone acetate) was associated with a reduction of pain during treatment. Counselling supported by ultrasound scanning was associated with reduced pain and improvement in mood. A multidisciplinary approach was beneficial for some outcome measures. Adhesiolysis was not associated with an improved outcome apart from where adhesions were severe. Sertraline was not beneficial. REVIEWER'S CONCLUSIONS: Further studies to confirm these observations are needed, together with full reporting of those studies which have been undertaken. Given the prevalence and health care costs associated with chronic pelvic pain in women, randomised controlled trials of other medical, surgical and psychological interventions are urgently required. PMID- 11034687 TI - Gonadotrophin therapy for ovulation induction in subfertility associated with polycystic ovary syndrome. AB - BACKGROUND: Approximately 15% of patients with PCOS remain anovulatory despite treatment with oral anti-oestrogen medications such as clomiphene citrate. In addition, about half of women with PCOS ovulating on anti-oestrogen treatment fail to conceive. Gonadotrophin stimulation is the next step in treatment for women who are "clomiphene resistant", however, results of gonadotrophin stimulation in women with PCOS are less successful. In PCOS associated with hypersecretion of LH, purified urinary follicle-stimulating hormone (u-FSH) preparations have theoretical advantages over the use of human menopausal gonadotrophin (hMG) preparations (containing both FSH and LH), but whether this claimed advantage extends into clinical practice remains uncertain. In addition, the use of gonadotrophin-releasing hormone analogues (GnRH-a) to produce pituitary desensitisation prior to ovulation induction in PCOS has been claimed to increase the success rates of treatment as well as reduce complications such as OHSS and multiple pregnancy. Gonadotrophin preparations have also been administered via different routes (intramuscular or subcutaneous), or using different stimulation regimens and protocols (step-up or standard) in an attempt to improve efficacy. OBJECTIVES: To determine the effectiveness of urinary derived gonadotrophins as ovulation induction agents in patients with PCOS trying to conceive. In particular, to assess the effectiveness of (1) different gonadotrophin preparations, (2) the addition of a gonadotrophin-releasing hormone agonist (GnRH-a) to gonadotrophin stimulation and (3) different modalities of gonadotrophin administration. SEARCH STRATEGY: The search strategy to identify RCTs consisted of (1) the Group's Specialised Register of Controlled Trials using the search strategy developed for the Menstrual Disorders and Subfertility Group as a whole (see the Review Group details for more information), (2) additional specific electronic Medline searches and (3) bibliographies of identified studies and narrative reviews. SELECTION CRITERIA: RCTs in which urinary-derived gonadotrophins were used for ovulation induction in patients with primary or secondary subfertility attributable to PCOS. DATA COLLECTION AND ANALYSIS: Twenty three RCTs were identified, 9 of which were excluded from analysis. The data were extracted independently by 2 authors. The following criteria were assessed: (1) the methodological characteristics of the trials, (2) the baseline characteristics of the studied groups and (3) the outcomes of interest: pregnancy rate (per cycle), ovulation rate (per cycle), miscarriage rate (per pregnancy), multiple pregnancy rate (per pregnancy), overstimulation rate (per cycle) and ovarian hyperstimulation syndrome (OHSS) rate (per cycle). Where suitable, meta analysis was performed using Peto's OR with 95% CI with the fixed effect Mantel Haentszel equation. MAIN RESULTS: (1) A reduction in the incidence of OHSS with FSH compared to hMG in stimulation cycles without the concomitant use of a GnRH-a (OR 0.20; 95% CI 0.08-0.46) and (2) a higher overstimulation rate when a GnRH-a is added to gonadotrophins (OR 3.15; 95% CI 1.48-6.70). REVIEWER'S CONCLUSIONS: Although 14 RCTs were included in this review, few dealt with the same comparisons, all were small to moderate size and their methodological quality was generally poor. Any conclusions, therefore, remain tentative as they are based on a limited amount of data and will require further RCTs to substantiate them. In none of the comparisons was there a significant improvement in pregnancy rate but this may be due to the lack of power (i.e. insufficient patients randomised to demonstrate a significant difference between treatments). There was a trend towards better pregnancy rates with the addition of a GnRH-a to gonadotrophin stimulation and these interventions warrant further study. Despite theoretical advantages, urinary-derived FSH preparations did not improve pregnancy rates when compared to traditional and cheaper hMG preparations; their only demonstrable benefit was a reduced risk of OHSS in cycles when administered without the concomitant use of a GnRH-a. No conclusions can be drawn on miscarriage and multiple pregnancy rates due to insufficient reporting of these outcomes in the trials. PMID- 11034688 TI - General versus spinal/epidural anaesthesia for surgery for hip fractures in adults. AB - BACKGROUND: The majority of hip fracture patients are treated surgically, requiring anaesthesia. OBJECTIVES: To compare different types of anaesthesia for surgical repair of hip fractures (proximal femoral fractures) in adults. This is primarily regional (spinal or epidural) anaesthesia versus inhalation general anaesthesia, but also includes ketamine anaesthesia versus inhalation general anaesthesia. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, Medline, selected orthopaedic and anaesthetic journals and conference proceedings, and reference lists of relevant articles. Date of the most recent search: August 1999. SELECTION CRITERIA: Randomised and quasi randomised trials comparing different methods of anaesthesia for hip fracture surgery in skeletally mature persons. Trials comparing the use of local nerve blocks are not considered in this review. Neither are trials using different types of drugs or techniques with one type of anaesthesia. The primary outcome was mortality. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, using a nine item scale, and extracted data. The other two reviewers independently checked these results. Wherever appropriate and possible, results were pooled. MAIN RESULTS: Sixteen trials, involving 2191 patients, which compared regional anaesthesia with general anaesthesia, were included. All trials had methodological flaws. Regional anaesthesia was associated with a decreased mortality at one month (53/781(6.8%) versus 78/826(9.4%)) of borderline statistical significance (relative risk 0.72, 95% confidence interval 0.51 to 1.00)). The results for three month mortality were not statistically significant, although the confidence interval does not exclude the possibility of a clinically relevant reduction (86/726 (11.8%) versus 98/765 (12.8%), relative risk 0.92, 95% confidence interval 0.71 to 1.21). The reduced numbers at one year, coming exclusively from two studies, preclude any useful conclusions for long term mortality (80/354 (22.6%) versus 78/372 (21.0%), relative risk 1.07, 95% confidence interval 0.82 to 1.41). Regional anaesthesia was associated with a tendency to a longer operation (weighted mean difference 4.8 minutes, 95% confidence interval 1.1 to 8.6 minutes), and a reduced risk of deep venous thrombosis (39/129 (30%) versus 61/37(76%); relative risk 0.64, 95% confidence interval 0.48 to 0.86), although this conclusion is insecure due to possible selection bias in the subgroups in whom this outcome was measured. No other statistically significant differences in outcome were identified. There was insufficient evidence to draw any conclusions from a further two included trials, involving a total of 100 patients, which compared other types of anaesthesia. REVIEWER'S CONCLUSIONS: Regional anaesthesia and general anaesthesia appear to produce comparable results for most of the outcomes studied. Regional anaesthesia may reduce short-term mortality but no conclusions can be drawn for longer term mortality. PMID- 11034689 TI - Psychotherapy for bulimia nervosa and binging. AB - BACKGROUND: Bulimia nervosa and like syndromes, such as binge eating disorder, are common in young Western women. A specific psychotherapy, cognitive behaviour therapy (CBT) has been developed for the treatment of bulimia nervosa. Other psychotherapies, some from a different theroretical framework, some modifications of CBT are also used. OBJECTIVES: The review aims to evaluate the psychotherapeutic treatments for those with binge eating syndromes, that have been tested in randomised controlled trials. Specifically, CBT therapy is compared with waiting list or a non-treatment group, any other psychotherapy, CBT in a "pure self-help" form and CBT augmented by exposure and response therapy. As well, the review aims to evaluate the evidence for the efficacy of other psychotherapies when compared to a no treatment control group and to evaluate the evidence for the efficacy of other psychotherapies when compared to a 'placebo' therapy. SEARCH STRATEGY: Handsearch of The International Journal of Eating Disorders since its first issue; database searches of MEDLINE, EXTRAMED, EMBASE, PSYCHLIT, CURRENT CONTENTS, LILACS, SCISEARCH, The Cochrane Collaboration Controlled Trials Register and the Cochrane Depression, Anxiety and Neurosis Group Database of Trials; citation list searching and personal approaches to authors communication are used. SELECTION CRITERIA: All studies that have tested any form of psychotherapy for adult patients with non-purging bulimia nervosa, binge eating disorder and/or EDNOS of a bulimic type, and which have applied a randomised controlled and standardized outcome methodology, are sought for the purpose of this review. DATA COLLECTION AND ANALYSIS: Data are entered into a spreadsheet programme, and into the REVMAN analysis program. Relative risk analyses are conducted of binary outcome data. The relative risk analysis is used rather than the odds ratio as the outcome measures proposed are not measuring a rare event (such as death) and the total number of studies is small. Standardized mean difference analyses are conducted of continuous variable outcome data, as the continuous outcome measures are not consistent across studies. Sensitivity analyses are conducted of a number of measures of trial quality. Data were not reported in such a way to permitsubgroup analyses, but the effect of treatment on depressive symptoms, psychosocial and/or interpersonal functioning, general psychiatric symptoms and weight is examined where possible. Chi-square tests for homogeneity are done, at 5% level of significance, using a fixed effects model. Funnel plots to evaluate presence of publication bias are completed and available in a text file upon request. MAIN RESULTS: To date, more than 1360 trials have been generated by searching and 63 trials have been evaluated in detail. Because of a relatively high number of exclusions (n=12) the trial inclusion criteria were broadened to include those with non-blinded outcome assessment, providing 25 trials for analyses. Because of incomplete published and available data, at best up to 10 studies had data available for any single analysis. The maximum number of total patients included in a single analysis is 543. The majority of studies (21) evaluate patients with bulimia nervosa of a purging type. CBT is superior to waiting list controls with respect to abstinence from binge eating (RR 0.64 CI.53 .78). CBT just fails to be significantly superior to other psychotherapies with respect to abstinence from binge eating (RR.79, CI.61-1.04). CBT in a full or less intensive form is not significantly superior to CBT in a pure self-help form. Augmentation of CBT with exposure therapy is not more effective than CBT alone. Non CBT-psychotherapies also have significantly greater abstinence rates in comparisons with wait-list controls, but there is a paucity of such studies (RR 0.67, CI.56-.81, n=3 studies). Funnel plots suggest a bias towards publication of positive outcome studies only. REVIEWER'S CONCLUSIONS: There is small body of evidence for the efficacy of cognitive-behaviour therapy in bulimia nervosa and similar syndromes, but the quality of trials is very variable (e.g. the majority are not blinded) and sample sizes are often very small. More trials are needed, particularly for binge eating disorder and other EDNOS syndromes, and evaluating other psychotherapies and less intensive psychotherapies. PMID- 11034690 TI - Carnitine supplementation of parenterally fed neonates. AB - BACKGROUND: Carnitine, a quaternary amino acid, plays an important role in the oxidation of long chain fatty acids. Both breast milk and infant formulas contain carnitine. However, it is not routinely provided in parenteral nutrition solutions. Non supplemented parenterally fed infants have very low tissue carnitine levels. The clinical significance of this is uncertain. Carnitine deficiency may be an etiological factor in the limited ability of premature babies to utilize parenteral lipid. In vitro studies have suggested that fatty acid oxidation is impaired when the tissue carnitine levels fall below 10% of normal. Therefore relative carnitine deficiency may impair fatty acid oxidation, thus reducing the available energy and impairing growth. OBJECTIVES: The primary aim of this review is to determine whether carnitine supplementation of parenterally fed neonates will improve weight gain. The secondary aims are to determine the effect on lipid tolerance and ketogenesis. SEARCH STRATEGY: Computerised searches were carried out by both reviewers. Searches were made of Medline, Embase, The National Research Register (UK), the Cochrane Controlled Trials Register and expert informants. The MeSH headings used were carnitine and parenteral nutrition. SELECTION CRITERIA: Only randomised trials were considered. Trials were included if they involved carnitine supplementation alone, parenterally fed newborn infants, and measured at least one outcome of interest (weight gain, plasma fatty acids, plasma triglycerides, quantity of lipid tolerated, respiratory quotient or beta hydroxybutyrate levels). DATA COLLECTION AND ANALYSIS: The two reviewers searched the literature separately and reached a consensus for inclusion of trials. Data were extracted and evaluated by the two reviewers independently of each other. Authors were contacted if possible to clarify or provide missing data. MAIN RESULTS: Fourteen studies were identified, six met the selection criteria. The results of the review are limited by the fact that the studies were generally short term and studied different outcomes. One study examined short term and long term weight gain, three reported only short term weight gain, three reported biochemical results in response to a short lipid challenge, and two reported results obtained during normal parenteral nutrition. Among infants supplemented with carnitine, there was no evidence of effect on weight gain, lipid utilization or ketogenesis. REVIEWER'S CONCLUSIONS: We found no evidence to support the routine supplementation of parenterally fed neonates with carnitine. PMID- 11034691 TI - Antimalarials for treating rheumatoid arthritis. AB - OBJECTIVES: To assess the short-term efficacy and toxicity of antimalarials for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register, Medline and Embase up to and including August 2000. We also carried out a handsearch of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing antimalarials against placebo in patients with RA DATA COLLECTION AND ANALYSIS: Data abstraction was carried out independently by two reviewers. The same two reviewers using a validated checklist (Jadad 1996) assessed the methodological quality of the RCTs and CCTs. Rheumatoid arthritis outcome measures were extracted from the publications for the 6-month endpoint. The pooled analysis was performed using standardized mean differences for joint counts, pain and global assessments. Weighted mean differences were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout. MAIN RESULTS: We found four trials, with 300 patients randomized to hydrochloroquine and 292 to placebo. Only trials evaluating hydroxychloroquine could be pooled in the analysis. A statistically significant benefit was observed when hydroxychloroquine was compared to placebo. The standardized mean differences for the various outcome measures ranged from -0.33 to -0.52, and were statistically significant. Statistically significant differences were also observed for ESR. Overall withdrawals and withdrawals due to lack of efficacy were significantly more frequent in the placebo group. No differences were observed in withdrawals due to toxicity. REVIEWER'S CONCLUSIONS: Hydroxychloroquine appears to be efficacious for the treatment of RA. Its overall effect appears to be moderate, but its low toxicity profile should be considered when treating patients with RA. PMID- 11034692 TI - Fully intermittent dosing with drugs for tuberculosis. AB - BACKGROUND: The number of people infected with tuberculosis continues to rise world-wide. Rifampicin-containing treatment regimens can achieve high cure rates. Intermittent drug treatment delivered in the community has the potential to improve adherence to treatment. OBJECTIVES: The objective of this review was to compare the effectiveness of rifampicin-containing short-course chemotherapy regimens, given two or three times a week, with similar regimens given daily in patients with pulmonary tuberculosis. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, the Cochrane Controlled Trials Register, Medline, and reference lists of articles. We contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of any multi drug regimen containing rifampicin in patients with confirmed pulmonary tuberculosis. Treatment had to be given up to three times a week for up to nine months, with any initial daily dosing period not more than one month, and was compared to daily dosing throughout for the same period. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial eligibility and quality. MAIN RESULTS: One trial involving 399 patients was included. The trial compared treatment three times per week with daily treatment for six months. There was no difference in cure rate (198 out of 199 people in the intermittent group compared to all 200 in the daily group), but 5 patients relapsed in the group receiving intermittent therapy compared to one in the group receiving the daily regimen. REVIEWER'S CONCLUSIONS: There is not enough evidence to assess the equivalence of effect between fully intermittent, rifampicin-containing short-course chemotherapy and similar daily therapy in patients with pulmonary tuberculosis. Larger randomised studies are required to establish the equivalence of fully intermittent, short-course chemotherapy, with daily regimens. PMID- 11034693 TI - Vaccines for preventing cholera. AB - BACKGROUND: Oral cholera vaccines are newer alternatives to the parenteral vaccines which have been thought to confer only moderate and short-term immunity. OBJECTIVES: The objective of this review was to assess the effect of cholera vaccines in preventing cases of cholera and preventing deaths. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group trials register, Medline, Embase and reference lists of articles. We handsearched the journal Vaccine, contacted researchers in the field and manufacturers. SELECTION CRITERIA: Randomised and quasi-randomised studies comparing cholera vaccines (killed or live) with placebo, control vaccines or no intervention, or comparing types, doses or schedules of cholera vaccine. We included adults and children irrespective of immune status or special risk category. DATA COLLECTION AND ANALYSIS: Data extraction and assessment of trial quality was done independently by two reviewers. MAIN RESULTS: Twenty-five trials were included. Eighteen efficacy trials of relatively good quality, testing parenteral and oral killed whole cell vaccines and involving over 2.6 million adults, children and infants were included. No randomised efficacy trials of live vaccines were available and therefore this review is restricted to killed vaccines only. Eleven safety trials have been conducted for both types of killed whole cell vaccines and have involved 9,342 people. For killed whole cell vaccines compared to placebo, the relative risk of contracting cholera at 12 months was 0.49, 95% confidence interval 0.41 to 0.59 (random effects model). This translates to an efficacy of 51%, 95% confidence interval 41% to 59%. Both parenteral and oral administration were relatively efficacious, but significant protection extended into the third year for oral killed whole cell vaccines. Children under 5 were only protected for up to a year, while older children or adults were protected for up to three years. Parenteral killed whole cell vaccines were associated with increased systemic and local adverse effects compared to placebo, while oral killed whole cell vaccines were not. REVIEWER'S CONCLUSIONS: Cholera killed whole cell vaccines appear to be relatively effective and safe. Protection against cholera appears to persist for up to two years following a single dose of vaccine, and for three to four years with an annual booster. PMID- 11034694 TI - Intravenous fluids for abdominal aortic surgery. AB - BACKGROUND: Surgery on the abdominal aorta, for aneurysmal and occlusive disease is a major undertaking which requires intensive support and fluid management. Blood products are often used, but the major fluid replacement is with crystalloids or colloids. There has been controversy for many years over which fluid is optimal and a number of studies have examined this subject, without any systematic review. OBJECTIVES: The objective of this review was to determine the effectiveness of different non-blood replacement fluids used in surgery on the abdominal aorta with a view to identifying the optimal fluid for use in such surgery. SEARCH STRATEGY: All publications describing (or which might describe) randomised controlled trials of non-blood replacement fluids in abdominal aortic surgery were sought using the search strategy described by the Cochrane Review Group on Peripheral Vascular Diseases. This strategy includes hand searching of relevant medical journals and extensive MEDLINE and EMBASE searches. In addition, trials have been identified from searches of references included in those trials already retrieved. SELECTION CRITERIA: Randomised controlled trials assessing the effects of one or more specific non-blood fluids used for replacement therapy in operations on, and confined to, the abdominal aorta. DATA COLLECTION AND ANALYSIS: Data were extracted to pre-prepared tables and then entered into the Review Manager software where statistical analysis and descriptive subjective analysis were performed. MAIN RESULTS: Nine trials, involving 412 patients were included. Patients undergoing aortic surgery had various physiological parameters measured before and after their operation (cardiac, respiratory, biochemical, haematological and protein). Ten fluids were studied: Ringer lactate, 5% dextrose in Ringer lactate, 5% dextrose in 0.45% saline, 5% dextrose in water, 1.8% saline, human albumin solution in Ringer lactate, human albumin solution in water, 5% dextrose with human albumin solution, Dextran 60, Hetastarch. Patients were randomised to fluid type. This review demonstrates that no single fluid has been shown to affect any outcome measure significantly more than any other across a range of outcome measures. However, each trial compared different fluids, and each fluid has not been compared against all others. The death rate in these studies was 2.9% (12 patients). REVIEWER'S CONCLUSIONS: Further studies are required, with sufficient sample size and power, to draw any further conclusions. There are no studies examining the effects of combination fluid therapy. PMID- 11034695 TI - Lecithin for dementia and cognitive impairment. AB - BACKGROUND: Alzheimer's disease sufferers have been found to have a lack of the enzyme responsible for converting choline into acetylcholine within the brain. Lecithin is a major dietary source of choline, so extra consumption may reduce the progression of dementia. OBJECTIVES: To determine the efficacy of lecithin in the treatment of dementia or cognitive impairment. SEARCH STRATEGY: The Cochrane Dementia and Cognitive Impairment Group Register of Clinical Trials has been searched, as have the electronic databases MEDLINE, EMBASE, Psychlit, ISI and Current Contents. Reference lists and relevant books have been examined. SELECTION CRITERIA: All unconfounded, randomized trials comparing lecithin with placebo in a treatment period longer than one day, in patients with dementia of the Alzheimer type, vascular dementia, mixed vascular and Alzheimer's disease, unclassified or other dementia or unclassified cognitive impairment not fulfilling the criteria for dementia are eligible for inclusion. DATA COLLECTION AND ANALYSIS: Data were extracted by two independent reviewers and cross-checked. Meta-analyses were performed when more than one trial provided data on a comparable outcome on sufficiently similar patients. Random effects analyses were performed whenever heterogeneity between results appeared to be present. Standardised mean difference were used due do the use of different scales and periods of treatment. Odds ratios for dichotomous data were pooled using the Mantel-Haenszel or DerSimonian and Laird methods. MAIN RESULTS: Twelve randomized trials have been identified involving patients with Alzheimer's disease (265 patients), Parkinsonian dementia (21 patients) and subjective memory problems (90 patients). No trials reported any clear clinical benefit of lecithin for Alzheimer's disease or Parkinsonian dementia. Few trials contributed data to meta analyses. The only statistically significant result was in favour of placebo for adverse events, based on one trial, which appears likely to be a spurious result. A dramatic result in favour of lecithin was obtained in a trial of subjects with subjective memory problems. REVIEWER'S CONCLUSIONS: Evidence from randomized trials does not support the use of lecithin in the treatment of patients with dementia. A moderate effect cannot be ruled out, but results from the small trials to date do not indicate priority for a large randomized trial. PMID- 11034696 TI - Antidepressant plus benzodiazepine for major depression. AB - BACKGROUND: Anxiety frequently coexists with depression. There is no systematic review to show if adding benzodiazepines to antidepressants can bring about any advantage over antidepressants alone in the treatment of depression, although such a combination prescription appears to be widely practiced worldwide OBJECTIVES: To determine whether, among adult patients with major depression, adding benzodiazepines to antidepressants brings about any benefit in terms of symptomatic recovery or side-effects in the short term (less than 8 weeks) and long term (more than 2 months), in comparison with treatment by antidepressants alone. SEARCH STRATEGY: Electronic searches of MEDLINE, EMBASE, International Pharmaceutical Abstracts, Biological Abstracts, LILACS, PsycLit, the Cochrane Library and the trial register of the Cochrane Depression, Anxiety and Neurosis Group (January 1972 to December 1998), combined with hand searching, reference searching, SciSearch and personal contacts. SELECTION CRITERIA: All randomized controlled trials that compared combined antidepressant-benzodiazepine treatment with antidepressant alone for adult patients with major depression (Feighner criteria, RDC, DSM-III, DSM-III-R, DSM-IV or ICD-10). Exclusion criteria are: antidepressant dosage lower than 100 mg of imipramine or its equivalent daily and duration of trial shorter than 4 weeks. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the eligibility and quality of the studies. Two reviewers independently extracted the data. Standardized weighted mean differences and relative risks were estimated with random effects model. The dropouts were assigned the least favorable outcome. Two sensitivity analyses examined the effect of this assumption as well as the effect of including medium quality studies. Three a priori subgroup analyses were performed with regard to the patients with or without comorbid anxiety and with regard to the types of benzodiazepines tested. MAIN RESULTS: Aggregating nine studies with a total of 679 patients, the combination therapy group was 37% (95%CI: 19 to 51%) less likely to drop out than the antidepressant alone group. The intention-to-treat analysis showed that the former were 63% (18 to 127%) to 38% (15 to 66%) more likely to show response (defined as 50% or greater reduction in the depression scale from baseline) up to 4 weeks. REVIEWER'S CONCLUSIONS: The potential benefits of adding a benzodiazepine to an antidepressant must be balanced judiciously against possible harms including development of dependence and accident proneness, on the one hand, and against continued suffering following no response and drop-out, on the other. PMID- 11034697 TI - Treatment programmes for people with both severe mental illness and substance misuse. AB - BACKGROUND: Substance misuse in the context of severe mental illness can have detrimental effects. A variety of treatments exist, but the drive has been to provide programmes integrating treatment of both substance misuse and severe mental illness. Such programmes require additional resources and may require radical redesign of service delivery systems. OBJECTIVES: To evaluate the effectiveness of treatment programmes within psychiatric care for people with problems of both substance misuse and serious mental illness. SEARCH STRATEGY: Biological Abstracts (1985-1998), CINAHL (1982-1998), The Cochrane Library (Issue 3, 1998), The Cochrane Schizophrenia Group's Register of trials (August 1998), EMBASE (1980-1998), MEDLINE (1966-1998), PsycLIT (1974-1998) and Sociofile (1974 1998) were comprehensively searched. Citations of all trials were searched and further studies sought from published trials and their authors. SELECTION CRITERIA: All randomised trials of any programme of substance misuse treatment for people with serious mental illness and current problems of substance misuse. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were also independently extracted. For homogeneous dichotomous data the Peto odds ratio (OR), and 95% confidence intervals (CI) were calculated on an intention-to-treat basis. MAIN RESULTS: Six relevant studies, four of which were small, were identified. In general, the quality of design and reporting was not high. Clinically important outcomes such as relapse of severe mental illness, violence to others, patient or carer satisfaction, social functioning and employment were not reported. There is no clear evidence supporting an advantage of any type of substance misuse programme for those with serious mental illness over the value of standard care. No one programme is clearly superior to another. REVIEWER'S CONCLUSIONS: The problems posed by substance misuse in the context of severe mental illness will not go away. The current momentum for integrated programmes is not based on good evidence. Implementation of new specialist substance misuse services for those with serious mental illnesses should be within the context of simple, well designed controlled clinical trials. PMID- 11034698 TI - Corticosteroids for pulmonary sarcoidosis. AB - BACKGROUND: Pulmonary sarcoidosis is a common condition with an unpredictable course. Oral or inhaled steroids are widely used in its treatment, but there is no consensus about when and in whom therapy should be initiated, what dose should be given and for how long. Corticosteroids given for several months have deleterious side-effects so it is important to know whether they have any maintained benefit in pulmonary sarcoidosis. OBJECTIVES: To determine the randomised controlled trial (RCT) evidence for the benefit of corticosteroids (oral or inhaled) in the treatment of pulmonary sarcoidosis. SEARCH STRATEGY: The Cochrane Airways Group interstitial lung disease RCT register was searched using the terms: sarcoidosis and (steroid* OR corticosteroid* OR prednisolone OR prednisone OR beclomethasone OR budesonide OR fluticasone). Bibliographies of retrieved RCTs and reviews were searched for additional RCTs. Pharmaceutical companies and authors of identified RCTs were contacted for other published and unpublished studies. SELECTION CRITERIA: Two reviewers independently assessed full text articles for inclusion based upon the following criteria: the study had to be a RCT or controlled clinical trial in adults with histological evidence of pulmonary sarcoidosis. DATA COLLECTION AND ANALYSIS: Study quality was assessed and data extracted independently by two reviewers. Outcomes were analysed as continuous and dichotomous outcomes, using standard statistical techniques. MAIN RESULTS: Nine RCTs were identified, two had insufficient data for any analysis. There were 516 patients in the five usable trials of oral steroids, and 66 patients in two trials of inhaled steroids. The oral steroid dose was equivalent to prednisolone 15-40 mg/day. The inhaled steroid was budesonide 0.8 - 1.2 mg/day. Outcomes were symptoms, chest X-ray (CXR) changes, lung function and global scores (a combination of all three outcomes). Oral steroids improved the CXR over 6-24 months. Two studies showed no improvement in lung function, in another there was an improvement in diffusing capacity in the treated group. Global scores improved in patients with Stage 2 and 3 disease but not with Stage 1 disease. There were no data on side-effects. Inhaled steroids had no effect on CXR. In one study diffusing capacity improved. In another, symptoms improved at the end of six months of treatment. REVIEWER'S CONCLUSIONS: Oral steroids improved the chest X-ray and a global score of CXR, symptoms and spirometry over 6-24 months., but there is little evidence of an improvement in lung function. There are no data beyond two years to indicate whether oral steroids have any modifying effect on long-term disease progression. Oral steroids are indicated for patients with Stage 2 and 3 disease with moderate - severe or progressive symptoms or CXR changes. The available data provide no guidance for the management of this disease after two years. Short term (less than six months) of inhaled steroids may improved symptoms, perhaps in patients who mainly have cough. PMID- 11034699 TI - Reality orientation for dementia. AB - BACKGROUND: Reality Orientation (RO) was first described as a technique to improve the quality of life of confused elderly people, although its origins lie in an attempt to rehabilitate severely disturbed war veterans, not in geriatric work. It operates through the presentation of orientation information (eg time, place and person-related) which is thought to provide the person with a greater understanding of their surroundings, possibly resulting in an improved sense of control and self-esteem. There has been criticism of RO in clinical practice, with some fear that it has been applied in a mechanical fashion and has been insensitive to the needs of the individual. There is also a suggestion that constant relearning of material can actually contribute to mood and self-esteem problems. There is often little consistent application of psychological therapies in dementia services, so a systematic review of the available evidence is important in order to identify the effectiveness of the different therapies. Subsequently, guidelines for their use can be made on a sound evidence base. OBJECTIVES: To assess the evidence of effectiveness for the use of Reality Orientation (RO) as a classroom-based therapy on elderly persons with dementia. SEARCH STRATEGY: Computerised databases were searched independently by 2 reviewers entering the terms 'Reality Orientation, dementia, control, trial or study'. Relevant web sites were searched and some hand searching was conducted by the reviewer. Specialists in the field were approached for undocumented material, and all publications found were searched for additional references. SELECTION CRITERIA: All randomised controlled trials (RCTs), and all controlled trials with some degree of concealment, blinding or control for bias (second order evidence) of Reality Orientation as an intervention for dementia were included. The criteria for inclusion/exclusion involved systematic assessment of the quality of study design and the risk of bias, using a standard data extraction form. A measure of cognitive and/or behavioural change was needed. DATA COLLECTION AND ANALYSIS: Data were extracted independently by both reviewers, using a previously tested data extraction form. Authors were contacted for data not provided in the papers. Psychological scales measuring cognitive and behavioural changes were examined. MAIN RESULTS: 6 RCTs were entered in the analysis, with a total of 125 subjects (67 in experimental groups, 58 in control groups). Results were divided into 2 subsections: cognition and behaviour. Change in cognitive and behavioural outcomes showed a significant effect in favour of treatment. REVIEWER'S CONCLUSIONS: There is some evidence that RO has benefits on both cognition and behaviour for dementia sufferers. Further research could examine which features of RO are particularly effective. It is unclear how far the benefits of RO extend after the end of treatment, but and it appears that a continued programme may be needed to sustain potential benefits. PMID- 11034700 TI - Reminiscence therapy for dementia. AB - BACKGROUND: Reminiscence Therapy (RT) has been defined as vocal or silent recall of events in a person's life, either alone, or with another person or group of people. It typically involves group meetings, at least once a week, in which participants are encouraged to talk about past events, often assisted by aids such as photos, music, objects and videos of the past. There is, often, little consistent application of psychological therapies in dementia services. A number of these 'therapies' were greeted with enthusiasm by health care practitioners in under stimulating care environments. They were expected to work miracles and their 'failure' to do this has led to their widespread disuse. A systematic review of the available evidence is important in order to identify the effectiveness of the different therapies. Subsequently, guidelines for their use can be made on a sound evidence base. OBJECTIVES: RT involves groups of elderly people talking of past events, assisted by aids such as videos, pictures and archives, as a means of communicating and reflecting upon their life experiences. The objective of the review is to assess the effects of RT for dementia. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE, PSYCHLIT, EMBASE, OMNI, BIDS, Dissertation Abstracts International, SIGLE and reference lists of relevant articles up to 1998, and we contacted specialists in the field. We also searched relevant Internet sites and we hand searched Aging and Mental Health, the Gerontologist, Journal of Gerontology, Current Opinion in Psychiatry, Current Research in Britain: Social Sciences, British Psychological Society conference proceedings and Reminiscence database. SELECTION CRITERIA: Randomised controlled trials and quasi-randomised trials of RT for dementia in elderly people. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Two trials are included in the review, but only one trial with 15 participants had extractable data. The results were statistically non-significant for both cognition and behaviour. REVIEWER'S CONCLUSIONS: No firm conclusions could be reached regarding the effectiveness of RT for dementia. The review highlighted the urgent need for more systematic research in the area. PMID- 11034701 TI - Drugs versus placebo for dysthymia. AB - OBJECTIVES: Dysthymia is a depressive disorder of chronic nature but of less severity than major depression, which depressive symptoms are more or less continuous for at least two years. The aim of this review was to conduct a systematic review of all RCTs comparing drugs and placebo for dysthymia. SEARCH STRATEGY: Electronic searches of Cochrane Library, EMBASE, MEDLINE, PsycLIT, Biological Abstracts and LILACS; reference searching; personal communication; conference abstracts; unpublished trials from the pharmaceutical industry; book chapters on the treatment of depression. SELECTION CRITERIA: The inclusion criteria for all randomised controlled trials were that they should focus on the use of drugs versus placebo for dysthymic patients. Exclusion criteria were: non randomised, mixed major depression/ dysthymia (trials not providing separate data) and depression secondary to other disorders (e.g. substance abuse). DATA COLLECTION AND ANALYSIS: The reviewers extracted the data independently. In order to achieve an intention-to-treat analysis, when trials failed to report it was assumed that people who died or dropped out had no improvement. Authors of relevant trials were contacted for additional and missing data. Absence of treatment response as defined by authors was the main measure of outcome used. Relative Risks (RR) and 95% confidence intervals (CI) of dichotomous data were calculated with the Random Effects Model. Where possible, number needed to treat (NNT) and number needed to harm (NNH) were estimated, taking the reciprocal of the absolute risk reduction. MAIN RESULTS: Currently the review includes 15 trials. Similar results were obtained in terms of efficacy for different groups of drugs, such as tricyclic (TCA), selective serotonin reuptake inhibitors (SSRI), monoamine oxidase inhibitors (MAOI) and other drugs (sulpiride, amineptine, and ritanserin). The pooled RR for absence of treatment response was 0.68 (95% CI 0.59-0.78) for TCA and the NNT was 4.3 (95% CI 3.2-6.5). SSRIs showed similar RR for this outcome: 0.64 (95% CI 0.55-0.74), the NNT being 4.7 (95% CI 3.5-6.9). Concerning MAOIs, the RR was 0.59 (95% CI 0.48-0.71) and the NNT was 2.9 (95% CI 2.2-4.3). Other drugs (amisulpride, amineptine and ritanserin) showed similar results in terms of absence of treatment response. Using more stringent criteria for improvement - full remission - the results were unchanged. Patients treated on TCA were more likely to report adverse events, compared with placebo. REVIEWER'S CONCLUSIONS: Drugs are effective in the treatment of dysthymia with no differences between and within class of drugs. Tricyclic antidepressants are more likely to cause adverse events and dropouts. As dysthymia is a chronic condition, there remains little information on quality of life and medium or long-term outcome. PMID- 11034702 TI - Cyclophosphamide for treating rheumatoid arthritis. AB - OBJECTIVES: To assess the short-term effects of cyclophosphamide for the treatment of rheumatoid arthritis. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's Register, the Cochrane Controlled Trials Register (issue 3, 2000), Medline and Embase up to and including August 2000. We also carried out a handsearch of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing oral cyclophosphamide against placebo (or an active drug at a dosage considered to be ineffective) in patients with rheumatoid arthritis. DATA COLLECTION AND ANALYSIS: Data abstraction was carried out independently by two reviewers. The same two reviewers using a validated checklist (Jadad 1996) assessed the methodological quality of the RCTs and CCTs. Rheumatoid arthritis outcome measures were extracted from the publications for baseline and end-of-study. The pooled analysis was performed using standardized mean differences (SMDs) for joint counts. Weighted mean differences (WMDs) were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout. MAIN RESULTS: A total of 70 patients were included in the pooled analysis of two trials, 31 receiving cyclophosphamide. A statistically significant benefit was observed for cyclophosphamide when compared to placebo for tender and swollen joint scores: SMDs were -0.57 and -0.59 respectively. The difference in ESR also favoured the active drug but did not reach statistical significance (-12 mm, 95%CI: -26 to 2.5). One trial reported the number of patients developing new or worse erosions: the OR for cyclophosphamide compared to placebo was 0.17 (95% CI: 0.05 to 0.57). Patients receiving placebo were six times more likely to discontinue treatment because of lack of efficacy than patients receiving cyclophosphamide. Withdrawals from adverse reactions were higher in the cyclophosphamide group (Odds ratio=2.9), although this difference was not statistically significant. Side effects from cyclophosphamide included hemorrhagic cystitis, nausea, vomiting, leucopenia, thrombocytopenia, alopecia, amenorrhea and herpes zoster infections. REVIEWER'S CONCLUSIONS: Cyclophosphamide appears to have a clinically and statistically significant benefit on the disease activity of patients with RA, similar to some disease modifying antirheumatic drugs (DMARDs) such as antimalarials or sulfasalazine, but lower than methotrexate. Toxicity however is severe, limiting its use given the low benefit-risk ratio compared to other antirheumatic agents. PMID- 11034703 TI - Therapeutic ultrasound for venous leg ulcers. AB - BACKGROUND: Ultrasound therapy is commonly used with the aim of improving the healing of chronic wounds such as pressure ulcers and venous leg ulcers. OBJECTIVES: To assess the effectiveness of therapeutic ultrasound in the treatment of venous leg ulcers. SEARCH STRATEGY: The Cochrane Wounds Group search strategy was used (see Scope) to search for randomised controlled trials (RCTs) of therapeutic ultrasound for the treatment of venous leg ulcers. SELECTION CRITERIA: Randomised controlled trials comparing therapeutic ultrasound with sham ultrasound, or other (standard) treatment DATA COLLECTION AND ANALYSIS: Results of searches were scrutinised by one reviewer (and checked by a second) to identify possible RCTs and full reports of these were obtained. Details of eligible studies were extracted and summarised using a data extraction sheet. Attempts were made to obtain missing data by contacting authors. Data extraction was checked by a second reviewer. MAIN RESULTS: A total of seven eligible RCTs were identified. Four trials compared ultrasound therapy with sham ultrasound, three trials compared ultrasound therapy with standard treatment. None of the trials found a difference in healing rates between any of the therapies, though it is noteworthy that the direction of treatment effect was consistently in favour of ultrasound (though this did not reach significance in the individual studies). REVIEWER'S CONCLUSIONS: There is no good evidence of a benefit of ultrasound therapy in the healing of venous leg ulcers. PMID- 11034704 TI - Donepezil for mild and moderate Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease is the most common cause of dementia in older people. One of the aims of therapy is to inhibit the breakdown of a chemical neurotransmitter, acetylcholine, by blocking the relevant enzyme. This can be done by a group of chemicals known as cholinesterase inhibitors. However, some (like tacrine) are associated with adverse effects such as hepatotoxicity, but E2020 (donepezil, Aricept) is thought to be more specific in its action, and safer. OBJECTIVES: The objective of this review is to assess whether or not donepezil improves the well-being of patients with mild or moderate Alzheimer's disease. SEARCH STRATEGY: The Cochrane Dementia and Cognitive Improvement Group specialized register was searched using the terms 'donepezil', 'E2020' and 'Aricept'. Members of the Donepezil Study Group and Eisai Inc were contacted. SELECTION CRITERIA: All unconfounded, double-blind, randomized controlled trials in which treatment with donepezil was compared with placebo for patients with Alzheimer's disease. DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer (JSB ), pooled where appropriate and possible, and the weighted mean differences or Peto odds ratios (95%CI) estimated. Where possible, intention-to treat (ITT) data were used. MAIN RESULTS: Eight trials are included, involving 2664 participants. The trials were of 12, 24 or 52 weeks duration in selected patients. Available outcome data cover domains including cognitive function and global clinical state, but data on several important dimensions of outcome are not available. For cognition there is a statistically significant improvement for both 5 and 10 mg/day of donepezil at 24 weeks compared to placebo (1.9 points on the ADAS-Cog scale, WMD 1.86, 95%CI -2.60 to -1.11; 2.9 points on the ADAS-Cog scale, WMD -2.91, 95% CI -3.65 to -2.16)and for 10mg/day donepezil compared to placebo at 52 weeks (1.7 MMSE points, 95% CI, -2.59 to -0.82). The results of three studies show some improvement in global clinical state (assessed by an independent clinician) in those treated with 5 and 10mg/day of donepezil compared with placebo at 12 and 24 weeks. The patients' own ratings of their Quality of Life showed no benefit of donepezil compared with placebo. There were significantly more withdrawals before the end of treatment from the 10mg/day (but not the 5mg/day) donepezil group compared with placebo which may have resulted in some overestimation of beneficial changes at 10mg/day A variety of adverse effects were recorded, with more incidents of nausea, vomiting, diarrhoea and anorexia in the 10mg/day group compared with placebo and the 5mg/day group, but very few patients left a trial as a direct result of the intervention. REVIEWER'S CONCLUSIONS: In selected patients with mild or moderate Alzheimer's disease treated for periods of 12, 24 or 52 weeks, donepezil produced modest improvements in cognitive function and study clinicians rated global clinical state more positively in treated patients. No improvements were present on patient self assessed quality of life and data on many important outcomes are not available. The practical importance of these changes to patients and carers is unclear. PMID- 11034705 TI - Rivastigmine for Alzheimer's disease. AB - BACKGROUND: Alzheimer's disease (AD) is the commonest cause of dementia affecting older people. One of the therapeutic strategies aimed at ameliorating the clinical manifestations of Alzheimer's disease is to enhance cholinergic neurotransmission in relevant parts of the brain by the use of cholinesterase inhibitors to delay the breakdown of acetylcholine released into synaptic clefts. Tacrine, the first of the cholinesterase inhibitors to undergo extensive trials for this purpose, was associated with significant adverse effects including hepatotoxicity. Several other cholinesterase inhibitors, including rivastigmine, with superior properties in terms of specificity of action and low risk of adverse effects, have now been introduced. Rivastigmine has received approval for use in 60 countries including all member States of the European Union and the USA. OBJECTIVES: To determine the clinical efficacy and safety of rivastigmine for patients with dementia of Alzheimer's type. SEARCH STRATEGY: The Cochrane Controlled Trials Register (April 2000) the Cochrane Dementia and Cognitive Improvement Group Register of Clinical Trials (July 2000), other electronic databases and other sources of reports were searched. SELECTION CRITERIA: All unconfounded, double-blind, randomized trials in which treatment with rivastigmine was administered to patients with dementia of the Alzheimer's type for more than two weeks and its effects compared with those of placebo in a parallel group of patients. DATA COLLECTION AND ANALYSIS: One reviewer (JSB) applied study selection criteria, assessed the quality of studies and extracted data. MAIN RESULTS: Seven trials, involving 3370 participants, were included. Use of rivastigmine in high doses was associated with statistically significant benefits on several measures. High-dose rivastigmine (6 to 12 mg daily) was associated with a 2.1 point improvement in cognitive function on the ADAS-Cog score compared with placebo (weighted mean difference -2.09, 95% confidence interval -2.65 to -1.54, on an intention-to-treat basis) and a 2.2 point improvment in activities of daily living assessed on the Progressive Deterioration Scale (weighted mean difference -2.15, 95% confidence interval 3.16 to -1.13, on an intention-to-treat basis) at 26 weeks. Fewer patients were graded as having severe dementia at 26 weeks (55% of patients taking rivastigmine compared with 59% on placebo; odds ratio 0.78, 95% confidence interval 0.64 to 0.94). At lower doses (4 mg daily or lower) differences were in the same direction but were statistically significant only for cognitive function. There were statistically significantly higher numbers of events of nausea, vomiting, diarrhoea, anorexia, headache, syncope, abdominal pain and dizziness among patients taking high-dose rivastigmine than among those taking placebo. There was some evidence that adverse events might be less common with more frequent, smaller doses of rivastigmine. REVIEWER'S CONCLUSIONS: Rivastigmine appears to be beneficial for people with mild to moderate Alzheimer's disease. In comparisons with placebo, improvements were seen in cognitive function, activities of daily living, and severity of dementia with daily doses of 6 to 12 mg. Adverse events were consistent with the cholinergic actions of the drug. Further resarch is desirable on dosage (frequency and quanitity) in a search for ways to minimize adverse effects. This review has not examined economic data. PMID- 11034706 TI - Hip protectors for preventing hip fractures in the elderly. AB - BACKGROUND: Hip fracture in the elderly is usually the result of a simple fall. Hip protectors have been advocated as a means to reduce the risk of sustaining a fracture in a fall on the hip. OBJECTIVES: To determine if external hip protectors reduce the incidence of hip fractures in elderly persons following a fall. SEARCH STRATEGY: The Cochrane Musculoskeletal Injuries Group trials register, MEDLINE, and reference lists of relevant articles were searched, and identified trialists contacted. Date of the most recent search: July 2000. SELECTION CRITERIA: All randomised or quasi-randomised controlled trials comparing the use of hip protectors with a control group. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, using a ten item scale, and extracted data. Additional information was sought from all trialists. Wherever appropriate and possible, the data are presented graphically. MAIN RESULTS: Six randomised trials involving 1752 participants were included within the review. All studies involved elderly people in nursing homes or residential care, three within the Scandinavian countries, one in Japan, one in the United Kingdom and one in Australia. The two largest studies involving 1409 participants randomised by nursing home or nursing home ward rather than by the individual (cluster randomisation). One study of 141 individuals was primarily a compliance study. Summation of results from the other five studies gave an occurrence of hip fractures of 16/660 (2.4%) for those allocated to wear hip protectors, against 63/951 (6.6%) to those not allocated to wear protectors. However due to the large number of participants allocated by cluster randomisation it was not possible to demonstrate conclusively that this difference between groups was statistically significant. Only one of the 16 hip fractures that occurred in the individuals allocated to wear hip protectors occurred whilst the protector was worn. No significant adverse effects of the hip protectors were reported but compliance, particularly in the long term, was poor. REVIEWER'S CONCLUSIONS: Hip protectors appear to reduce the risk of hip fracture within a selected population at high risk of sustaining a hip fracture. However, this conclusion is based on five trials of low to moderate quality. As two used cluster randomisation, pooling of data was limited. The generalisation of the results is unknown beyond high-risk populations. Results from eleven ongoing trials may clarify this situation. Acceptability by users of the protectors remains a problem, due to discomfort and practicality. PMID- 11034707 TI - Therapeutic ultrasound for pressure sores. AB - BACKGROUND: Pressure sores have been recorded as occurring in 4-10% of patients admitted to a UK District General Hospital (the precise rate depends on case-mix) and in an unknown proportion of patients in the community. They represent a major burden of sickness and reduced quality of life for patients and their carers, and are costly to health service providers. Pressure sores can be treated by using wound dressings, relieving pressure on the wound, by treating concurrent conditions which may delay healing, and by the use of physical therapies such as electrical stimulation, laser therapy and ultrasound. OBJECTIVES: To assess the effectiveness of the use of therapeutic ultrasound in the treatment of pressure sores. SEARCH STRATEGY: The Cochrane Wounds Groups search strategy was used (see Scope) to search for randomised controlled trials (RCTs) of therapeutic ultrasound for the treatment of pressure sores up to December 1999. SELECTION CRITERIA: Randomised controlled trials comparing therapeutic ultrasound with sham ultrasound or other (standard) treatment. DATA COLLECTION AND ANALYSIS: Results of searches were scrutinised by one reviewer to identify possible RCTs and full reports of these were obtained. Details of eligible studies were extracted and summarised using a data extraction sheet. Attempts were made to obtain missing information by contacting authors. Data extraction was checked by a second reviewer. Meta-analysis was used to combine the results of trials where the interventions and outcome measures were sufficiently similar. MAIN RESULTS: A total of 3 eligible RCTs were identified. Two RCTs compared ultrasound therapy with sham and the third compared a combination of ultrasound and ultraviolet light with laser and with standard treatment. Neither of the two RCTs comparing ultrasound with sham found a significant difference in healing rates. The trials were pooled, in the absence of significant heterogeneity. There was no evidence of benefit associated with the use of ultrasound in the treatment of pressure sores. In the three-arm comparison there was a significant increase in the weekly healing rates associated with the ultrasound/ultraviolet combination compared with laser but no statistically significant difference between ultrasound/ultraviolet and control. REVIEWER'S CONCLUSIONS: The results suggest no apparent evidence of a benefit of ultrasound therapy in the treatment of pressure sores. However the possibility of a beneficial or a harmful effect cannot be ruled out due to the small number of trials with methodogical limitations and small numbers of participants. PMID- 11034708 TI - Dietary marine fatty acids (fish oil) for asthma. AB - BACKGROUND: Epidemiological studies suggest that a diet high in marine fatty acids (fish oil) may have beneficial effects on inflammatory conditions such as rheumatoid arthritis and possibly asthma. OBJECTIVES: 1. To determine the effect of marine n-3 fatty acid (fish oil) supplementation in asthma. 2. To determine the effect of a diet high in fish oil in asthma. SEARCH STRATEGY: The Cochrane Airways Review Group register was search using the terms: marine fatty acids OR diet OR nutrition OR fish oil OR eicosapentaenoic acid OR EPA. Bibliographies of retrieved trials were searched and fish oil manufacturers contacted. SELECTION CRITERIA: Randomised controlled trials in patients with asthma more than two years of age were included. The study duration had to be in excess of 4 weeks. Double blind trials were preferred, but single-blind and open trials were also reviewed for possible inclusion. Three reviewers read each paper, blind to its identity. Decisions concerning inclusion were made by simple majority. Quality assessment was performed by all three reviewers independently. DATA COLLECTION AND ANALYSIS: The only comparison possible was between marine n-3 fatty acid supplementation and placebo. There were insufficient trials to examine dietary manipulation alone. MAIN RESULTS: Eight randomised controlled trials conducted between 1986 and 1998 satisfied the inclusion criteria. Six were of parallel design and two were cross-over studies. Seven compared fish oil with placebo whilst one compared high dose vs low dose marine n-3 fatty acid supplementation. None of the included studies reported asthma exacerbations, health status or hospital admissions. There was no consistent effect on any of the analyzable outcomes: FEV1, peak flow rate, asthma symptoms, asthma medication use or bronchial hyper reactivity. The single study performed in children also combined dietary manipulation with fish oil supplementation and showed improved peak flow and reduced asthma medication use. There were no adverse events associated with fish oil supplements. Updated Search conducted August 2000. No new trials were found. REVIEWER'S CONCLUSIONS: There is little evidence to recommend that people with asthma supplement or modify their dietary intake of marine n-3 fatty acids (fish oil) in order to improve their asthma control. Equally, there is no evidence that they are at risk if they do so. PMID- 11034709 TI - Inhaled short acting beta2-agonist use in asthma: regular vs as needed treatment. AB - BACKGROUND: Inhaled beta2-adrenergic agonists delivered by inhalation are very widely used in asthma. There has been much controversy of the use and possible consequences of the use of these agents for regular, as opposed as-needed use in asthma. OBJECTIVES: This review to assessed the clinical trial evidence to test whether using regular use of short-acting beta2-agonists reduced asthma control and pulmonary function; worsened symptoms, airway reactivity and quality of life; and increased the rate of exacerbations. SEARCH STRATEGY: A search was carried out of the Cochrane Airways Group "Asthma and Wheez* RCT" register using the terms: regular AND [beta agonist OR bronchodilator OR salbutamol OR albuterol Or terbutaline OR isoproterenol OR reproterol OR fenoterol]. Bibliographies of existing trials were searched and primary trial authors and pharmaceutical companies were approached for additional trials. SELECTION CRITERIA: Randomised controlled trials in which the short-acting beta2-agonist was given regularly in the experimental group, together with an inhaled bronchodilator for relief of symptoms ('rescue use'). The control group consisted of matching placebo inhaled regularly, with an inhaled bronchodilator for as-needed 'rescue use'. DATA COLLECTION AND ANALYSIS: Data were extracted and quality assessments were made by both reviewers. Parallel group and cross-over trials were analysed separately. Where possible data were pooled using a fixed effects model. MAIN RESULTS: Over 800 abstracts were identified, following a review, 60 papers were requested for full assessment by both reviewers. 34 trials from 30 papers met the entry criteria. Data from 31 outcomes were analyzable. There was little difference between the treatments for nearly all outcomes. In cross-over studies, evening peak flow was better with regular treatment, weighted mean difference (WMD) 13.1 l/min (95% confidence interval 24.3, 1.9). In contrast, the FEV1 was better with as-needed treatment (WMD 157 ml (95% CI: 123, 192). Bronchial hyper reactivity was slightly better in the as-needed group, standardised mean difference 0.23, 95% CI: 0.52, 1.12. REVIEWER'S CONCLUSIONS: These results support current guidelines. There is little advantage in using short-acting beta2-agonists regularly, and potentially some small clinical disadvantage. PMID- 11034710 TI - Aspirin for vascular dementia. AB - BACKGROUND: For patients with a diagnosis of vascular dementia there is evidence that aspirin is widely prescribed - in one study, completed by geriatricians and psychiatrists in the UK, 80% of patients with cognitive impairment (with vascular risk factors) were prescribed aspirin. However, a number of queries remain unanswered: Is there convincing evidence that aspirin benefits patients with vascular dementia? Does aspirin affect cognition or improve prognosis? In addition, does the risk of cerebral or gastric haemorrhage outweigh any benefit? OBJECTIVES: To assess the evidence of effectiveness of the use of aspirin for vascular dementia. SEARCH STRATEGY: Computerized databases were searched independently by two reviewers. In addition, relevant websites were searched and some journals were handsearched. Specialists in the field were approached for unpublished material and any publications found were searched for additional references. The most recent search for trials was carried out in February 2000. SELECTION CRITERIA: All randomized controlled trials investigating the effect of aspirin for vascular dementia are included. Inclusion/exclusion of studies comprised systematic assessment of the quality of study design and the risk of bias. DATA COLLECTION AND ANALYSIS: Data were extracted independently by both reviewers, using a previously tested data extraction form and, where required, authors were contacted for data not provided in the papers. The aim is to evaluate data recorded via tools assessing cognitive and behavioural changes along with mortality, morbidity and institutionalization data. MAIN RESULTS: No trials are eligible for inclusion in this review. REVIEWER'S CONCLUSIONS: There is no evidence that aspirin is effective in treating patients with a diagnosis of vascular dementia. Further research is needed to assess the effect of aspirin on cognition, and on other outcomes such as behaviour, and quality of life. At present there is no evidence relating to other queries about the use of aspirin for dementia (these are described in the Background section of this review). The most recent search for references to relevant research was carried out in February 2000, but no new evidence was found. PMID- 11034711 TI - Antibiotics for preventing leptospirosis. AB - BACKGROUND: Leptospirosis is an infectious disease transmitted by animals. Death occurs in about five per cent of the patients. In clinical practice, doxycycline is widely used for prevention. OBJECTIVES: To evaluate the effectiveness and safety of any antibiotic regimen versus placebo or other antibiotic regimens in the prophylaxis of leptospirosis. SEARCH STRATEGY: The sources used were: EMBASE, LILACS, MEDLINE, SCISEARCH, The Cochrane Controlled Trials Register, The Cochrane Hepato-Biliary Group Controlled Trials Register, bibliographies of published papers, and personal communication with authors. There were no language or date restrictions in any of the searches. SELECTION CRITERIA: STUDIES: All randomised clinical trials in which antibiotics were used as prophylactic regimen for leptospirosis. PARTICIPANTS: People potentially exposed to leptospirosis, such as people in endemic areas during the rainy season, health professionals and other professionals with high risk of infection. INTERVENTION: Any antibiotic regimen compared with a control group (placebo or another antibiotic regimen). OUTCOMES: Infection (primary outcome) and adverse events (secondary outcome). DATA COLLECTION AND ANALYSIS: Data were independently extracted and methodological quality of each trial was assessed by two reviewers as well as cross-checked. Details of the randomisation (generation and concealment), blinding, and the number of patients lost to follow-up were recorded. The results of each trial were summarised on an intention-to-treat basis in 2 x 2 tables for each outcome. MAIN RESULTS: Two trials comparing doxycycline with placebo met the inclusion criteria. We did not find trials comparing doxycycline versus other antibiotics, or other antibiotics versus placebo. One of the trials had excellent methodological quality. In the other trial, the allocation concealment process, generation of allocation sequence, and blinding methods were not described. Of the 1022 participants enrolled, 509 were treated with doxycycline and 513 with placebo. Of these, 940 participants were soldiers included in one trial. The patients assigned to the antibiotics group compared with the ones assigned to the placebo group showed: Symptomatic, verified leptospirosis: 0.6% (3/509) versus 4.9% (25/ 513); risk difference (random effects model) -4.1%, 95% confidence interval -5.9% to -2.3%. Number needed-to-treat 24 (95% confidence interval 17 to 43). Adverse effects: 3% (13/469 participants) versus 0.2% (1/471 participants); random effects model 2.6%, 95% confidence interval 1.0% to 4.1%. Number needed-to harm 39 (95% confidence interval 25 to 100). REVIEWER'S CONCLUSIONS: Prophylaxis of leptospirosis may be achieved by administrating doxycycline to soldiers training in endemic areas with a high risk of exposure to leptospirosis. Whether these findings apply to other scenarios or not remains to be proven. PMID- 11034712 TI - Antidepressants for depression in medical illness. AB - OBJECTIVES: To determine whether antidepressants are clinically effective and acceptable for the treatment of depression in people who also have a physical illness. SEARCH STRATEGY: Medline, Cochrane Library Trials Register and Cochrane Depression and Neurosis Group Trials Register were all systematically searched, supplemented by hand searches of two journals and reference searching. SELECTION CRITERIA: All relevant randomised trials comparing any antidepressant drug (as defined in the British National Formulary) with placebo or no treatment, in patients of either sex over 16, who have been diagnosed as depressed by any criterion, and have a specified physical disorder (for example cancer, myocardial infarction). "Functional" disorders where there is no generally agreed physical pathology (e.g. irritable bowel syndrome) were excluded. The main outcome measures are numbers of individuals who recover/improve at the end of the trial and, as a proxy for treatment acceptability, numbers who complete treatment. DATA COLLECTION AND ANALYSIS: Data was extracted independently by the reviewers onto data collection forms and differences settled by discussion. MAIN RESULTS: 18 studies were included, covering 838 patients with a range of physical diseases (cancer 2, diabetes 1, head injury 1, heart 1, HIV 5, lung 1, multiple sclerosis 1, renal 1, stroke 3, mixed 2). Depression was diagnosed clinically in 3 studies, otherwise by structured interview or checklist. Only 5 studies described how they performed randomisation. 1 study compared drug with no treatment, and the rest with placebo: all of the latter said they were double blind. 6 studies used SSRIs, 3 atypical antidepressants, and the remainder tricyclics. Patients treated with antidepressants were significantly more likely to improve than those given placebo (13 studies, OR 0.37, 95% CI 0.27-0.51) or no treatment (1 study, OR 3.45, 95% CI 11.1-1.10). About 4 patients would need to be treated with antidepressants to produce one recovery from depression which would not have occurred had they been given placebo (NNT 4.2, 95% CI 3.2-6.4). Most antidepressants (tricyclics and SSRIs together, 15 trials ) produced a small but significant increase in dropout (OR 1.66, 95% CI 1.14-2.40. NNH 9.8, 95% CI 5.4 42.9). The "atypical" antidepressant mianserinproduced significantly less dropout than placebo. Only 2 studies used numerical scales designed to measure effects on function and quality of life; in HIV (Karnofsky scale), drug was better than no treatment; in lung disease (Sickness Impact Profile), drug was not significantly different from placebo. Only 7 studies reported looking for changes in the physical disease. Antidepressants produced no change in immune function in HIV relative to placebo (2 studies) or no treatment (1 study). Relative to placebo, antidepressants produced no change in cardiovascular function in heart disease, in respiratory function in lung disease, or in vital signs or laboratory tests in cancer (1 study each). Nortriptyline produced worse control in diabetes. Trends towards tricyclics being more effective than SSRIs, but also more likely to produce dropout were noted, but these are based on non-randomised comparisons between trials. REVIEWER'S CONCLUSIONS: The review provides evidence that antidepressants, significantly more frequently than either placebo or no treament, cause improvement in depression in patients with a wide range of physical diseases. About 4 patients would need to be treated with antidepressants to produce one recovery from depression which would not have occurred had they been given placebo (NNT 4.2, 95% CI 3.2-6.4). Antidepressants seem reasonably acceptable to patients, in that about 10 patients would need to be treated with antidepressants to produce one dropout from treatment which would not have occurred had they been given placebo (NNH 9.8, 95% CI 5.4-42.9). The evidence is consistent across the trials, apart from 2 trials in cancer, where the "atypical" antidepressant mianserin produced significantly less dropout than placebo. Trends towards tricyclics being more effective than SSRIs, but also more likely to produce dropout were noted, but these are based on non-randomised comparisons between trials. Problems with the evidence include most of the trials' use of observers, rather than patients, to decide on improvement, and concentration mainly on symptoms rather than function and quality of life. There is also a possibility of undetected negative trials. Nevertheless, the review provides evidence that use of antidepressants should at least be considered in those with both physical illness and depression. Regarding diagnosis, the existence of a cheap and readily available treatment for depression should encourage detailed assessment of persistent low mood in the physically ill. PMID- 11034713 TI - Corticosteroids or ACTH for acute exacerbations in multiple sclerosis. AB - BACKGROUND: Corticosteroids are often used to improve the rate of recovery from acute exacerbation in multiple sclerosis (MS) patients. However, it is still unclear just how relatively effective these agents are and the type of drug, optimal dose, frequency, duration of treatment and route of administration are unknown. OBJECTIVES: The object of this review was to determine the efficacy and safety of corticosteroids or ACTH in reducing the short and long term morbidity from MS. Moreover, we wished to examine from indirect comparisons if the effect of corticosteroids is different according to different doses and drugs, routes of administration, length of treatment. SEARCH STRATEGY: A search strategy developed for the Cochrane MS Group (last searched: June 1999) completed with handsearching and personal contacts with trialists and pharmaceutical companies was used. SELECTION CRITERIA: All randomised, double-blind, unconfounded trials comparing corticosteroids or ACTH to placebo in patients with MS, treated for acute exacerbations, without any age or severity restrictions, were evaluated. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected articles for inclusion, assessed trials' quality and extracted the data. A third reviewer cross-checked them and disagreements were resolved by a joint discussion. MAIN RESULTS: Six trials contributed to this review; a total of 377 participants (199 treatment, 178 placebo) were randomised. The drugs analysed were methylprednisolone (MP) (four trials, 140 patients) and ACTH (two trials, 237 patients). Overall, MP or ACTH showed a protective effect against the disease getting worse or stable within the first five weeks of treatment (odds ratio[OR]=0.37, 95% confidence interval [CI] 0.24-0.57) with some but non significant greater effect for MP and intravenous administration. Short (five days) or long (15 days) duration of treatment with MP did not show any significant difference. Only one study (with 51 patients) reported data after one year of follow-up: no difference between oral MP and placebo in the prevention of new exacerbations or improvement in long term disability was detected. No data are available beyond one year of follow-up to indicate whether steroids or ACTH have any effect on long-term progression. One study reported that a short term treatment with high dose intravenous MP was not attended by adverse events. On the contrary, gastrointestinal symptoms and psychic disorders were significantly more common in the oral, high-dose MP than in the placebo group. Weight gain and edema were significantly more frequent in the ACTH group than in controls. REVIEWER'S CONCLUSIONS: We found evidence favouring the corticosteroid MP for acute exacerbation in MS patients. Data are insufficient to reliably estimate effect of corticosteroids on prevention of new exacerbations and reduction of long-term disability. Studies assessing long term risk/benefit and adverse effects of corticosteroids in MS patients are urgently needed. PMID- 11034714 TI - Anti-spasticity agents for multiple sclerosis. AB - BACKGROUND: Spasticity is a common problem in MS patients causing pain, spasms, loss of function and difficulties in nursing care. A variety of oral and parenteral medications are available. OBJECTIVES: To assess the absolute and comparative efficacy and tolerability of anti-spasticity agents in multiple sclerosis (MS) patients. SEARCH STRATEGY: Randomised controlled trials (RCTs) of anti-spasticity agents were identified using MEDLINE, EMBASE, bibliographies of relevant articles, personal communication, manual searches of relevant journals and information from drug companies. SELECTION CRITERIA: Double-blind, randomised controlled trials (either placebo-controlled or comparative studies) of at least seven days duration. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted data and the findings of the trials were summarised. Missing data were collected by correspondence with principal investigators. A meta-analysis was not performed due to the inadequacy of outcome measures and methodological problems with the studies reviewed. MAIN RESULTS: Twenty-three placebo-controlled studies (using baclofen, dantrolene, tizanidine, botulinum toxin, vigabatrin, prazepam and threonine) and thirteen comparative studies met the selection criteria. Only thirteen of these studies used the Ashworth scale, of which only three of the six placebo-controlled trials and none of the seven comparative studies showed a statistically significant difference between test drugs. Spasms, other symptoms and overall impressions were only assessed using unvalidated scores and results of functional assessments were inconclusive. REVIEWER'S CONCLUSIONS: The absolute and comparative efficacy and tolerability of anti-spasticity agents in multiple sclerosis is poorly documented and no recommendations can be made to guide prescribing. The rationale for treating features of the upper motor neurone syndrome must be better understood and sensitive, validated spasticity measures need to be developed. PMID- 11034715 TI - Naltrexone maintenance treatment for opioid dependence. AB - BACKGROUND: Despite widespread use of naltrexone maintenance in many countries for more than a decade, the evidence of its effects has not yet been systematically evaluated. OBJECTIVES: To evaluate the effects of naltrexone maintenance treatment in preventing relapse in opioid addicts after detoxification. SEARCH STRATEGY: We searched MEDLINE (1973-first year of naltrexone use in humans-July 2000), EMBASE (1974-July 2000), Cochrane Controlled Trials Register (Cochrane Library issue 2000, 3) and handsearched the "Bolletino per le Farmacodipendenze e l'Alcolismo" (1978 to 1997) and reference lists of relevant articles. We contacted pharmaceutical producers of naltrexone, authors and other Cochrane review groups. Date of most recent searches: July 2000. SELECTION CRITERIA: All controlled studies of naltrexone; treatment of heroin addicts after detoxification. DATA COLLECTION AND ANALYSIS: Reviewers evaluated data independently and analysed outcome measures taking into consideration adherence to and success of the study intervention. Data was extracted and analysed stratifying for the three categories of study quality. Where possible, meta-analysis was performed. MAIN RESULTS: Eleven studies met the criteria for inclusion in this review, even if not all of them were randomised. The methodological quality of the included study varied, but was generally poor. Meta analysis could be performed to a very low degree only, because the studies and their outcome measures were very heterogeneous. A statistically significant reduction of (re-)incarcerations was found for patients treated with naltrexone and behaviour therapy respect to those treated with behaviour therapy only. The other outcomes considered in the meta-analysis did not yield any significant results. Final conclusions on whether naltrexone treatment may be considered effective in maintenance therapy cannot be drawn from the clinical trials available so far. REVIEWER'S CONCLUSIONS: The available trials do not allow a final evaluation of naltrexone maintenance treatment yet. A trend in favour of treatment with naltrexone was observed for certain target groups (particularly people who are highly motivated), as has been previously described in the literature. PMID- 11034716 TI - Oral misoprostol for induction of labour. AB - BACKGROUND: Misoprostol is a synthetic prostaglandin which has been used to induce labour. Oral use of the drug misoprostol may be convenient, but an overdose could cause uterine hyperstimulation and precipitate labour which may be life-threatening for both mother and fetus. OBJECTIVES: The objective of this review was to assess the effects of oral misoprostol used for labour induction in women with a viable fetus. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched in May 2000. SELECTION CRITERIA: Randomised trials of oral misoprostol versus other methods, placebo or no treatment given to women with a viable fetus for labour induction. DATA COLLECTION AND ANALYSIS: This is one of a series of the Cochrane reviews of methods of cervical ripening and labour induction using standardised methodology. The Cochrane Pregnancy and Childbirth Group has developed a strategy to deal with the large volume and complexity of trial data related to labour induction. Data from all relevant trials are extracted centrally. They are incorporated into a series of primary reviews arranged by methods of induction of labour. The data from the primary reviews will be incorporated into secondary reviews. The secondary reviews are arranged by category of woman according to parity, membrane status and previous caesarean section. To avoid duplication of data in the primary reviews, the labour induction methods have been listed in a specific, hierarchical order. This review includes comparisons between oral misoprostol with only those methods above it on the list (placebo, vaginal prostaglandins, intracervical prostaglandins, oxytocin, amniotomy, oxytocin and amniotomy or vaginal misoprostol). MAIN RESULTS: One trial with 80 randomised women with prelabour rupture of membranes at term showed that, compared with placebo, oral misoprostol reduces the need for oxytocin infusion from 51 percent to 13 percent (relative risk 0.25, 95% confidence interval 0.1 to 0.6) and shortens delivery time by 8.7 hours (95% CI 6.0 to 11.3). Compared with vaginal prostaglandins, oral misoprostol showed no beneficial or harmful effects. However, only two trials were included with 957 randomised women in total. In six trials with 1042 randomised women that compared oral with vaginal misoprostol, oral misoprostol appeared to be less effective. More women in the oral misoprostol group did not achieve vaginal delivery within 24 hours of randomisation (54.2%) compared with 35.2% in the vaginal misoprostol group (relative risk 1.55, 95% confidence intervals 1.30 to 1.85). Caesarean section rate was 17.7% in the oral misoprostol group compared with 21.7% in the vaginal misoprostol group (relative risk 0.82, 95% confidence intervals 0.64 to 1.05). There was no difference in uterine hyperstimulation with fetal heart rate changes (9.5% versus 9.9%; relative risk 0.93, 95% confidence intervals 0.66 to 1.33). There were no reported cases of severe neonatal and maternal morbidity. REVIEWER'S CONCLUSIONS: Oral misoprostol may be an effective method for labour induction. However, the data on safety are lacking. It is possible that clinically effective oral regimens may have an unacceptably high incidence of complications such as uterine hyperstimulation and possibly uterine rupture. PMID- 11034717 TI - Adjuvant chemotherapy for localised resectable soft tissue sarcoma in adults. AB - BACKGROUND: Individually, randomised trilas have not shown conclusively whether adjuvant chemotherapy benefits adult patients with localised resectable soft tissue sarcoma. OBJECTIVES: Adjuvant chemotherapy aims to lessen the recurrence of cancer after surgery with or without radiotherapy. The objective of this review was to assess the effects of adjuvant chemotherapy in adults with resectable soft tissue sarcoma after such local treatment. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, UKCCCR Register of Cancer Trials, Physicians Data Query, EMBASE, MEDLINE and CancerLit. SELECTION CRITERIA: Randomised trials of adjuvant chemotherapy after local treatment in adults with localised resectable soft tissue sarcoma were included. Only trials in which accrual was completed by December 1992 were included. DATA COLLECTION AND ANALYSIS: Individual patient data were obtained. Accuracy of data and quality of randomisation and follow-up of trials was assessed. MAIN RESULTS: Fourteen trials of doxorubicin-based adjuvant chemotherapy involving 1568 patients were included. Median follow-up was 9.4 years. For local recurrence-free interval the hazard ratio with chemotherapy was 0.73 (95% Confidence Interval 0.56-0.94). For distant recurrence-free interval it was 0.70 (95% CI 0.57-0.85). For overall recurrence free survival it was 0.75 (95% CI 0.64-0.87). These correspond to significant absolute benefits of 6-10% at 10 years. For overall survival the hazard ratio of 0.89 (95% CI 0.76-1.03) was not significant but potentially represents an absolute benefit of 4% (95% CI -1 to 9) at 10 years. There was no consistent evidence of a difference in effect according to age, sex, stage, site, grade, histology, extent of resection, tumour size or exposure to radiotherapy. However, the strongest evidence of a beneficial effect on survival was shown in patients with sarcoma of the extremities. REVIEWER'S CONCLUSIONS: Doxorubicin-based adjuvant chemotherapy appears to significantly improve time to local and distant recurrence and overall recurrence-free survival in adults with localised resectable soft tissue sarcoma. There is some evidence of a trend towards improved overall survival. PMID- 11034718 TI - Intravenous or enteral loop diuretics for preterm infants with (or developing) chronic lung disease. AB - BACKGROUND: Lung disease in preterm infants is often complicated with lung edema. OBJECTIVES: The aim of this review was to assess the risks and benefits of administration of a diuretic acting on the loop of Henle (loop diuretic) in preterm infants with or developing chronic lung disease (CLD). Primary objectives were to assess changes in need for oxygen or ventilatory support and effects on long-term outcome, and secondary objectives were to assess changes in pulmonary mechanics and potential complications of therapy. SEARCH STRATEGY: We used the standard search method of the Cochrane Neonatal Review Group. We used the following keywords: [ or ] and , limited to and limited to or . We searched Medline (1966-1998), Embase (1974-1998) and the Cochrane Controlled Trials Register (CCTR) from the Cochrane Library (1998, issue 4). In addition, we hand searched several abstract books of national and international American and European Societies. SELECTION CRITERIA: We included in this analysis trials in which preterm infants with or developing chronic lung disease and at least 5 days of age were all randomly allocated to receive a loop diuretic either enterally or intravenously. Eligible studies needed to assess at least one of the outcome variables defined a priori for this systematic review. Primary outcome variables included important clinical outcomes, and secondary outcome variables included toxicity and pulmonary mechanics (e.g., lung compliance and airway resistance). DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study, using a form that was designed specifically for this review. Any disagreement was resolved by discussion. We combined parallel and cross-over trials and, whenever possible, transformed baseline and final outcome data measured on a continuous scale into change scores using Follmann's formula. MAIN RESULTS: The only loop diuretic used in the studies which met the selection criteria was furosemide. Most studies focused on pathophysiological parameters and did not assess effects on important clinical outcomes defined in this review, or the potential complications of diuretic therapy. In preterm infants < 3 weeks of age developing CLD, furosemide administration has either inconsistent effects or no detectable effect. In infants > 3 weeks of age with CLD, a single intravenous dose of 1 mg/kg of furosemide improves lung compliance and airway resistance for 1 hour. Chronic administration of furosemide improves both oxygenation and lung compliance. REVIEWER'S CONCLUSIONS: In preterm infants > 3 weeks of age with CLD, acute and chronic administration of furosemide improve lung compliance. Chronic administration of intravenous or enteral furosemide improves oxygenation. In view of the lack of data from randomized trials concerning effects on important clinical outcomes, routine or sustained use of systemic loop diuretics in infants with (or developing) CLD cannot be recommended based on current evidence. Randomized trials are needed to assess the effects of furosemide administration on survival, duration of ventilatory support and oxygen administration, length of hospital stay, potential complications and long-term outcome. PMID- 11034719 TI - Penicillamine for treating rheumatoid arthritis. AB - OBJECTIVES: To estimate the short-term effects of D-penicillamine for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register (issue 3, 2000) and Medline up to and including August 2000 and Embase from 1988 2000. We also carried out a handsearch of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials and controlled clinical trials comparing D-penicillamine against placebo in patients with rheumatoid arthritis. DATA COLLECTION AND ANALYSIS: The methodological quality of the trials was assessed independently by two reviewers (CS, EB) and checked by a third (MS) using a validated quality assessment tool (Jadad 1996). Rheumatoid arthritis outcome measures were extracted from the publications for the six-month endpoint and stratified according to D-penicillamine dosages: low (<500mg/day), moderate (500 to <1000mg/day) and high (1000 mg/day or greater). Data was abstracted by one reviewer and checked by a second (CS, MS). The pooled analysis was performed using the standardized mean difference for joint counts, pain and global assessments. The weighted mean difference was used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals and adverse reactions. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout, since no statistical heterogeneity was found. MAIN RESULTS: Six trials were identified, with 425 patients randomized to D-penacillamine and 258 to placebo. A statistically significant benefit was observed for D-penicillamine when compared to placebo for all three-dose ranges and for most outcome measures including: tender joint counts, pain, physician's global assessments and ESR. The standardized weighted mean differences between treatment and placebo in moderate doses were -0.51 [95% CI -0.88, -0.14] for tender joint counts, -0.56 (95% CI -0.87, -0.26) for pain and -0.97 (95% CI -1.25, -0.70) for global assessment. The difference for ESR was -10.6 mm/hr. Similar results were observed for the higher dose group. Total withdrawals were significantly higher in the moderate and high dosage D penicillamine groups (OR=1.63 and 2.13 respectively), mostly due to increased adverse reactions (OR = 2.60 and 4.95 respectively), including renal and hematological abnormalities. REVIEWER'S CONCLUSIONS: D-penicillamine appears to have a clinically and statistically significant benefit on the disease activity of patients with rheumatoid arthritis. Its efficacy appears to be similar to that of other disease modifying anti-rheumatic drugs (DMARDs), but with a significantly higher toxicity. Its effects on long-term functional status and radiological progression are not clear from this review. PMID- 11034720 TI - Azathioprine for treating rheumatoid arthritis. AB - OBJECTIVES: To assess the short-term effects of azathioprine for the treatment of rheumatoid arthritis (RA). SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register (issue 3, 2000), Medline up to and including August 2000 and Embase from 1988 to August 2000. We also conducted a handsearch of the reference lists of the trials retrieved from the electronic search. SELECTION CRITERIA: All randomized controlled trials and controlled clinical trials comparing azathioprine against placebo in patients with rheumatoid arthritis. DATA COLLECTION AND ANALYSIS: Data was extracted independently by two reviewers (CS, EB); disagreements were resolved by discussion or third party adjudication (MS). The same reviewers (CS, EB) assessed the methodological quality of the trials using a validated quality assessment tool. Rheumatoid arthritis outcome measures were extracted from the publications for the six-month endpoint. The pooled analysis was performed using standardized mean differences for joint counts, pain and functional status assessments. Weighted mean differences were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals and for adverse reactions. The 95% confidence intervals (95% CI) are presented. A chi square test was used to assess heterogeneity among trials. Fixed effects models were used throughout, since no statistical heterogeneity was found. MAIN RESULTS: Three trials with a total of 81 patients were included in the analysis. Forty patients were randomized to azathioprine and forty-one to placebo. A pooled estimate was calculated for two outcomes. A statistically significant benefit was observed for azathioprine when compared to placebo for tender joint scores. The standardized weighted mean difference between treatment and placebo was -0.98 (95% CI -1.45, -0.50). Withdrawals from adverse reactions were significantly higher in the azathioprine group OR=4.56 (95% CI 1.16, 17.85). REVIEWER'S CONCLUSIONS: Azathioprine appears to have a statistically significant benefit on the disease activity in joints of patients with RA. This evidence however is based on a small number of patients, included in older trials. Its effects on long-term functional status and radiological progression were not assessed due to lack of data. Toxicity is shown to be higher and more serious than that observed with other disease-modifying anti-rheumatic drugs (DMARDs). Given this high risk to benefit ratio, there is no evidence to recommend the use of azathioprine over other DMARDs. PMID- 11034721 TI - Antibiotics for preventing pneumonia in children with measles. AB - BACKGROUND: Measles causes more than a million deaths a year, of which most are children under five years of age who die from pneumonia. OBJECTIVES: The objective of this review was to assess the effects of antibiotics given to children with measles on reducing pneumonia or mortality, and to assess whether antibiotics should be given to all children with measles in communities with a high fatality rate. SEARCH STRATEGY: We searched MEDLINE (1966 - 1999), EMBASE (1980-1999) and the specialized trials register of the Acute Respiratory Infections Group in August 1999, and all relevant journals in the University of Melbourne medical library for the years 1935-46. SELECTION CRITERIA: Randomised or controlled trials of antibiotics for children with measles. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: Six trials with 1304 children were included. All but one of the trials were unblinded, and randomisation was either not described or was by alternate allocation. In four studies, the incidence of pneumonia in the control group was similar to that in the antibiotic prophylaxis group; in the other two studies, the incidence of pneumonia was unusually high in the control group so these children had a higher complication rate than the antibiotic group. Four of the 764 children given antibiotics died compared with one of the 637 controls. REVIEWER'S CONCLUSIONS: The quality of the trials reviewed was poor, and they provide very weak evidence for giving antibiotics to all children with measles. Available evidence suggests that antibiotics should be given only if a child has clinical signs of pneumonia or other evidence of sepsis. PMID- 11034722 TI - Aminosteroids for acute traumatic brain injury. AB - BACKGROUND: Traumatic brain injury is a leading cause of premature death and disability. Post-traumatic membrane lipid peroxidation has been proposed as one mechanism leading to secondary brain damage following head injury. Aminosteroids have been shown to inhibit lipid peroxidation in laboratory animals and have the potential to improve outcome following head injury. OBJECTIVES: To quantify the effectiveness and safety of aminosteroids in the treatment of acute traumatic brain injury. SEARCH STRATEGY: We searched the Cochrane Injuries Group trials register, The Cochrane Controlled Trials Register, MEDLINE and EMBASE. We contacted experts in the field and the company that manufactures tirilazad. SELECTION CRITERIA: We sought to identify all randomised controlled trials of aminosteroids versus placebo in the treatment of acute traumatic brain injury. Studies using a quasi random form of allocation, such as alternation, were excluded from the review. DATA COLLECTION AND ANALYSIS: One reviewer examined the electronic search results for reports of possibly relevant trials for retrieval in full. Two reviewers (IR and PA) applied the selection criteria independently to the trial report, with no disagreement. MAIN RESULTS: Two randomised controlled trials have examined the effect of the aminosteroid tirilazad mesylate on death and disability following head injury. To date, only the results of one of these trials are available for analysis. The risk of death in patients treated with tirilazad was almost identical to those given placebo RR=1.05 (95% confidence interval 0.86 to 1.29). The risk of death and severe disability in patients treated with tirilazad was again almost identical to those given placebo RR=1.07 (95% confidence interval 0.93 to 1.23). REVIEWER'S CONCLUSIONS: There is no evidence to support the routine use of aminosteroids in the management of traumatic head injury. On the basis of the existing evidence from randomised trials of aminosteroids in head injury it is not possible to refute the possibility of moderate but potentially clinically important benefits or harms. A further randomised controlled trial of tirilazad mesylate with 1156 participants has been completed, the results of which should become available in the near future. PMID- 11034723 TI - Corticosteroid therapy for nephrotic syndrome in children. AB - BACKGROUND: In nephrotic syndrome protein leaks from the blood to the urine through the glomeruli resulting in hypoproteinaemia and generalised oedema. About 70% of children experience a relapsing course with recurrent episodes of oedema and proteinuria. Children with untreated nephrotic syndrome frequently die from infections. The majority of children with nephrotic syndrome respond to corticosteroids. Corticosteroid usage has reduced the mortality rate in childhood nephrotic syndrome to around 3%, with infection remaining the most important cause of death. However corticosteroids have known adverse effects such as obesity, poor growth, hypertension, diabetes mellitus and osteoporosis. The original treatment schedules for childhood nephrotic syndrome were developed in an ad hoc manner. The optimal doses and durations of corticosteroid therapy that are most beneficial and least harmful have not been clarified. The aim of this systematic review is to assess the benefits and harms of corticosteroid therapy in treating children with nephrotic syndrome. OBJECTIVES: To determine the benefits and harms of different corticosteroid regimes in preventing relapse in children with steroid responsive nephrotic syndrome (SRNS). SEARCH STRATEGY: Published and unpublished randomised controlled trials were identified from the Cochrane Controlled Trials Register, Medline, Embase, reference lists of articles, abstracts from proceedings and contact with known investigators in the area. SELECTION CRITERIA: Randomised trials were included if they were carried out in children (aged three months to 18 years) in their initial or subsequent episode of SRNS, if they compared different durations, total doses or other dose strategies using prednisone or other corticosteroid agent and if they had outcome data at six months or more. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed all eligible studies for inclusion, assessed study quality and extracted data. The principle outcome measure was the number of children with and without relapse after six and 12-24 months. Secondary outcomes sought included the number of children who developed frequently relapsing nephrotic syndrome and adverse events. A random effects model was used to estimate summary effect measures (relative risk RR, risk difference RD) after testing for heterogeneity. Meta-regression was used to explore potential between-study differences due to the baseline risk of relapse, study quality and types of interventions used. MAIN RESULTS: Twelve trials were identified. A meta-analysis of five trials, which compared two months of prednisone with three months or more in the first episode, showed that the longer duration significantly reduced the risk of relapse at 12 - 24 months (relative risk 0.73; 95% CI 0.60,0.89) without an increase in adverse events. There was an inverse linear relationship (RR = 1.382 (SE 0.215) - 0.133 duration (SE 0.048); r2 = 0.66; p = 0.05) between the duration of treatment and risk of relapse. The number of children who became frequent relapsers and the mean relapse rate/patient/year were also significantly reduced without increase in serious adverse events. In children with frequently relapsing nephrotic syndrome, deflazacort was significantly more effective in maintaining remission than prednisone (RR 0.44; 95% CI 0.25, 0.78). REVIEWER'S CONCLUSIONS: From this meta-analysis of randomised controlled trials it can be concluded that children in their first episode of nephrotic syndrome should be treated for at least three months with an increase in benefit being demonstrated for up to seven months of treatment. In a population with a baseline risk for relapse following the first episode of 60% with two months of prednisone, daily prednisone for four weeks followed by alternate day therapy for six months would be expected to reduce the number of children experiencing a relapse by about 40%. In children who relapse frequently, deflazacort deserves further study. PMID- 11034724 TI - Local corticosteroid injection for carpal tunnel syndrome. AB - BACKGROUND: Carpal tunnel syndrome (CTS) is a clinical syndrome manifested by signs and symptoms of irritation of the median nerve at the level of the carpal tunnel in the wrist. Treatment of CTS can be surgical or non-surgical. Local corticosteroid injection for CTS has been previously studied but most studies have been either retrospective or uncontrolled. The effectiveness and duration of benefit of local corticosteroid injection for CTS remain unknown. OBJECTIVES: To evaluate the effectiveness of local steroid injection for carpal tunnel syndrome versus placebo injection or other non-surgical interventions in improving clinical outcome and to determine the length of symptom relief post injection. SEARCH STRATEGY: We searched the Cochrane Neuromuscular Disease Group register, MEDLINE, EMBASE and CINAHL. SELECTION CRITERIA: Studies using either a randomized or quasi-randomized methodology were eligible for inclusion. The studies included participants with the diagnosis of carpal tunnel syndrome and the treatment intervention was local corticosteroid injection. The primary outcome measure was clinical improvement after injection. DATA COLLECTION AND ANALYSIS: Three reviewers independently selected the trials to be included in the study. Studies were rated for their overall quality independently by the reviewers. Studies were compared for heterogeneity using the chi square statistic. Relative risks and 95% confidence intervals were calculated for each trial and summary relative risks and 95% confidence intervals (CI) were also calculated. MAIN RESULTS: We identified four randomized controlled trials studying local corticosteroid injection for the treatment of CTS. Two trials were excluded. One did not include clinical assessment as an outcome and the other did not provide patient outcomes, but only statistical values. Each of the remaining two trials had demonstrated clinical improvement of CTS at one month following local corticosteroid injection compared to placebo injection. The pooled relative risk (RR) favouring treatment was 3.62 (95% CI 1.94 to 6.73). REVIEWER'S CONCLUSIONS: Local corticosteroid injection for CTS provides greater clinical improvement in symptoms one month after injection compared to placebo. Symptom relief beyond one month compared to placebo has not been demonstrated. The effectiveness of local corticosteroid injection has not been compared to other non-surgical or surgical interventions for CTS in randomized controlled trials. PMID- 11034725 TI - Electrical stimulation for preventing and treating post-stroke shoulder pain. AB - BACKGROUND: Shoulder pain after stroke is common and disabling. The optimal management is uncertain, but electrical stimulation (ES) is often used to treat and prevent pain. OBJECTIVES: The objective of this review was to determine the efficacy of any form of surface ES in the prevention and / or treatment of pain around the shoulder at any time after stroke. SEARCH STRATEGY: We searched the Cochrane Stroke Review Group trials register and undertook further searches of MEDLINE, EMBASE and CINAHL. Contact was established with equipment manufacturers and centres that have published on the topic of ES. SELECTION CRITERIA: We considered all randomised trials that assessed any surface ES technique (functional electrical stimulation (FES), transcutaneous electrical nerve stimulation (TENS) or other), applied at any time since stroke for the purpose of prevention or treatment of shoulder pain. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials for inclusion, assessed trial quality and extracted the data. MAIN RESULTS: Four trials (a total of 170 subjects) fitted the inclusion criteria. Study design and ES technique varied considerably, often precluding the combination of studies. Population numbers were small. There was no significant change in pain incidence (Odds Ratio (OR) 0.64; 95% CI 0.19 to 2.14) or change in pain intensity (Standardised Mean Difference (SMD) 0.13; 95% CI -1.0 to 1.25) after ES treatment compared to control. There was a significant treatment effect in favour of ES for improvement in pain-free range of passive humeral lateral rotation (Weighted Mean Difference (WMD) 9.17; 95% CI 1.43 to 16.91). In these studies ES reduced the severity of glenohumeral subluxation (SMD -1.13; 95% CI -1.66 to -0.60), but there was no significant effect on upper limb motor recovery (SMD 0.24; 95% CI -0.14 to 0.62) or upper limb spasticity (WMD 0.05; 95% CI -0.28 to 0.37). There did not appear to be any negative effects of electrical stimulation at the shoulder. REVIEWER'S CONCLUSIONS: The evidence from randomised controlled trials so far does not confirm or refute that ES around the shoulder after stroke influences reports of pain, but there do appear to be benefits for passive humeral lateral rotation. A possible mechanism is through the reduction of glenohumeral subluxation. Further studies are required. PMID- 11034726 TI - Domiciliary oxygen for chronic obstructive pulmonary disease. AB - BACKGROUND: Domiciliary oxygen therapy has become one of the major forms of treatment for hypoxaemic chronic obstructive pulmonary disease (COPD) patients. OBJECTIVES: To determine the effect of domiciliary oxygen therapy on survival and quality of life in patients with COPD. SEARCH STRATEGY: Randomised controlled trials (RCTs) were identified using the Cochrane Airways Group COPD register using the search terms: home OR domiciliary AND oxygen. SELECTION CRITERIA: Any RCT in patients with hypoxaemia and COPD that compared long term domiciliary or home oxygen therapy with a control treatment. DATA COLLECTION AND ANALYSIS: Data extraction was performed independently by two reviewers. MAIN RESULTS: Five randomised controlled trials were identified. Data was aggregated from two trials of the treatment of nocturnal oxygen therapy in patients with mild to moderate COPD and arterial desaturation at night. Data could not be aggregated for the other three trials because of differences in trial design and patient selection. Nott 1980: continuous oxygen therapy versus nocturnal oxygen therapy: there was a significant improvement in mortality after 24 months (Peto odds ratio 0.45, 95% confidence interval 0.25 to 0.81). MRC 1981: domiciliary oxygen therapy versus no oxygen therapy: there was a significant improvement over five years in mortality in the group receiving oxygen therapy (Peto odds ratio 0.42, 95% confidence interval 0.18 to 0.98). In two studies of nocturnal oxygen versus no oxygen in patients with COPD and arterial desaturation at night: there was no difference in mortality between treated and non treated groups for either trial or when the trials were aggregated. In one study of long term oxygen versus no oxygen in moderate hypoxaemia: there was no effect on survival for up to three years of follow up. REVIEWER'S CONCLUSIONS: Long term oxygen therapy improved survival in a selected group of COPD patients with severe hypoxaemia (arterial PO2 less than 8.0 kPa). Long term oxygen did not appear to improve survival in patients with moderate hypoxaemia or in those with only arterial desaturation at night. PMID- 11034727 TI - Follow-up strategies for women treated for early breast cancer. AB - BACKGROUND: Follow-up examinations are commonly performed after primary treatment for women with breast cancer. They are used to detect recurrences at an early (asymptomatic) stage. OBJECTIVES: To assess the effectiveness of different policies of follow-up for distant metastases on mortality, morbidity and quality of life in women treated for early breast cancer. SEARCH STRATEGY: We searched the Breast Cancer Groups specialised register, the Cochrane Controlled Trials Register ( Cochrane Library Issue 4, 1999), MEDLINE (January 1975-September 1999) and EMBASE (1988-September 1999) using "Breast Neoplasms" and "follow-up". References from retrieved articles were also checked, as were the lists of presentations from recent breast cancer meetings. SELECTION CRITERIA: All randomised controlled trials (RCTs) assessing the effectiveness of different policies of follow-up after primary treatment were reviewed for inclusion. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and eligibility for inclusion in the review. Data were pooled in an individual patient data meta-analysis for the two RCTs testing the effectiveness of different follow-up schemes. Subgroup analyses by age, tumour size and lymph node status before primary treatment are also presented. MAIN RESULTS: Four RCTs involving 3204 women with early breast cancer (clinical stage I, II or III) have been included. Two RCTs involving 2563 women compared follow-up based on clinical visits and mammography with a more intensive scheme including radiological and laboratory tests. After pooling the data, no significant differences in overall survival (hazard ratio 0.96, 95% confidence interval 0.80 to 1.15) or disease free survival (hazard ratio 0.84, 95% confidence interval 0.71 to 1.00) emerged. No differences in overall survival and disease-free survival emerged in subgroup analyses according to patient age, tumour size and lymph node status before primary treatment. One RCT (296 women) compared follow-up performed by a hospital based specialist to follow-up performed by general practitioners. No significant differences in time to detection of recurrence and quality of life emerged. One RCT (196 women) compared regularly scheduled follow-up visits to less frequent visits restricted to the time of mammography. No significant differences emerged in interim use of telephone and frequency of GP's consultations. REVIEWER'S CONCLUSIONS: Follow-up programs based on regular physical examinations and yearly mammography alone appear to be as effective as more intensive approaches based on regular performance of laboratory and instrumental tests in terms of timeliness of recurrence detection, overall survival and quality of life. In one RCT, follow up care performed by general practitioners had comparable effectiveness to that delivered by hospital based specialists in terms of quality of life and time to detection of distant metastases. PMID- 11034728 TI - Preoperative radiotherapy for esophageal carcinoma. AB - BACKGROUND: The existing randomized evidence has failed to conclusively demonstrate the benefit or otherwise of preoperative radiotherapy in treating patients with potentially resectable esophageal carcinoma. OBJECTIVES: This meta analysis aimed to assess whether there is benefit from adding radiotherapy prior to surgery and whether or not any pre-defined patient subgroups benefit more or less from preoperative radiotherapy SEARCH STRATEGY: Medline and CancerLit searches were supplemented by information from trial registers and by hand searching relevant meeting proceedings and by discussion with relevant trialists, organisations and industry. The search strategy was run again in Medline, Embase and the Cochrane Library on 2nd May 2000, one year after original publication. No new trials were found. SELECTION CRITERIA: Trials were eligible for inclusion in this meta-analysis provided they randomized patients with potentially resectable carcinoma of the esophagus (of any histological type) to receive radiotherapy or no radiotherapy prior to surgery. Trials must have used a randomization method which precluded prior knowledge of treatment assignment and completed accrual by December 1993, to ensure sufficient follow-up by the time of the first analysis (September 1995). DATA COLLECTION AND ANALYSIS: A quantitative meta-analysis using updated data from individual patients from all properly randomized trials (published or unpublished) comprising 1147 patients (971 deaths) from five randomized trials. This approach was used to assess whether preoperative radiotherapy improves overall survival and whether it is differentially effective in patients defined by age, sex and tumour location. MAIN RESULTS: With a median follow-up of 9 years, in a group patients with mostly squamous carcinomas, the hazard ratio (HR) of 0.89 (95% CI 0.78-1.01) suggests an overall reduction in the risk of death of 11% and an absolute survival benefit of 3% at 2 years and 4% at 5 years. This result is not conventionally statistically significant (p=0.062). No clear differences in the size of the effect by sex, age or tumor location were apparent. REVIEWER'S CONCLUSIONS: Based on existing trials, there was no clear evidence that preoperative radiotherapy improves the survival of patients with potentially resectable esophageal cancer. These results indicate that if such preoperative radiotherapy regimens do improve survival, then the effect is likely to be modest with an absolute improvement in survival of around 3 to 4%. Trials or a meta-analysis of around 2000 patients (90% power, 5% significance level) would be needed to reliably detect such an improvement (from 15 to 20%). PMID- 11034729 TI - Exercise-based rehabilitation for coronary heart disease. AB - BACKGROUND: The burden of cardiovascular disease world-wide is one of great concern to patients and health care agencies alike. Circulatory diseases, including myocardial infarction (MI) and stroke, kill more people than any other disease. Cardiac rehabilitation aims to restore patients who have suffered myocardial infarction to optimal health through exercise only based rehabilitation or comprehensive cardiac rehabilitation (eg. smoking cessation advice, diet and counselling as well as exercise). Data from two published and widely cited meta-analyses (Oldridge 1988, O'Connor 1989) of over 4,000 patients each have demonstrated that patients randomised to exercise-based cardiac rehabilitation after MI have a statistically significant reduction in all-cause and cardiac mortality of about 20 to 25% compared to patients receiving conventional care. However, the trials included were small and often of poor methodological quality. Incomplete literature review methods may have resulted in publication bias thereby resulting in an over-estimate of the benefit of cardiac rehabilitation. The randomised controlled trials used in the reviews have focused almost exclusively on low-risk, middle-aged males post MI, thereby excluding women and the elderly. OBJECTIVES: To determine the effectiveness of exercise only rehabilitation and exercise in addition to other rehabilitation interventions (termed comprehensive cardiac rehabilitation) compared with usual care on the mortality, morbidity, health-related quality of life (HRQoL) and modifiable cardiac risk factors of patients with coronary heart disease. SEARCH STRATEGY: Electronic databases were searched for randomised controlled trials, using standardised trial filters, from the earliest date available to December 31st 1998. SELECTION CRITERIA: Men and women of all ages, in both hospital-based and community-based settings, who have had myocardial infarction, coronary artery bypass graft or percutaneous transluminal coronary angioplasty, or who have angina pectoris or coronary artery disease defined by angiography have been included. Studies involving participants following heart transplant, heart valve surgery or heart failure have been excluded. Follow up periods of less than 6 months were excluded. DATA COLLECTION AND ANALYSIS: Studies were selected independently by two reviewers, and data extracted independently. Authors were contacted where possible to obtain missing information. MAIN RESULTS: The current systematic review has allowed analysis of an increased number of patients from approximately 4500 in the earlier meta-analyses to 7683 (2582 in exercise only and 5101 in the comprehensive cardiac rehabilitation group). The quality of reporting overall was poor, with generally high losses to follow up. The pooled effect estimate for total mortality for the exercise only intervention shows a 27% reduction in all cause mortality (random effects model OR 0.73 (0.54, 0.98)). Similarly, comprehensive cardiac rehabilitation reduced all cause mortality compared to usual care, but to a lesser degree (OR 0.87 (0.71, 1.05)). Total cardiac mortality was reduced by 31% (random effects model OR 0.69 (0.51, 0.94)) and 26% (random effects model OR 0.74 (0.57, 0.96)) in the exercise only and comprehensive cardiac rehabilitation intervention groups respectively when compared to usual care. Neither intervention had any effect on the ocurrence of non-fatal myocardial infarction. There was a significant net reduction in total cholesterol in the comprehensive cardiac rehabilitation group (pooled WMD random effects model -0.57 mmol/l (-0.83, -0.31)), but not the exercise only rehabilitation group. Similarly, LDL was significantly reduced in the comprehensive cardiac rehabilitation group (pooled WMD random effects model -0.51 mmol/l (-0.82, -0.19). The effect of exercise only rehabilitation or comprehensive cardiac rehabilitation interventions on revascularisation rates, blood pressure or smoking behaviour could not be determined by this meta-analysis due to the small number of trials reporting these outcomes and heterogeneity between trials. It was not possible to combine the data from studies reporting HRQoL as an outcome. Eighteen different instruments were used to assess HRQoL in the 11 studies reporting it as an outcome. The data are presented qualitatively, only one trial reporting significant improvements with the intervention. REVIEWER'S CONCLUSIONS: Exercise-based cardiac rehabilitation appears to be effective in reducing cardiac deaths but the evidence base is weakened by poor quality trials. It is not clear from this review whether exercise only or a comprehensive cardiac rehabilitation intervention is more beneficial. The population studied in this review is still predominately male, middle aged and low risk. Identification of the ethnic origin of the participants was seldom reported. (ABSTRACT TRUNCATED) PMID- 11034730 TI - Developmental care for promoting development and preventing morbidity in preterm infants. AB - BACKGROUND: Preterm infants experience a range of morbidity related to the immaturity of their organ systems and to concurrent disease states. An unfavourable environment in the neonatal intensive care unit (NICU) may compound this morbidity. Modification of the environment could minimize the iatrogenic effects. Developmental care is a broad category of interventions designed to minimize the stress of the NICU environment. These interventions may include one or more elements such as control of external stimuli (vestibular, auditory, visual, tactile), clustering of nursery care activities, and positioning or swaddling of the preterm infant. Individual strategies have also been combined to form programs, such as the 'Neonatal Individualized Developmental Care and Assessment Program' (NIDCAP) (Als 1986). OBJECTIVES: In preterm infants, do developmental care interventions reduce neurodevelopmental delay, poor weight gain, length of hospital stay, length of mechanical ventilation, physiological stress and other clinically relevant adverse outcomes? SEARCH STRATEGY: The Neonatal Review Group search strategy was utilized. Searches were made of Medline from 1966 to July, 2000, and of CINAHL, The Cochrane Library, and conference and symposia proceedings in the English language from 1990 to July, 2000. A list of all relevant articles was sent to two experts in the field to identify any omissions or additional unpublished studies. SELECTION CRITERIA: Randomized trials in which elements of developmental care are compared to routine nursery care for infants < 37 weeks gestation and that measured clinically relevant outcomes. Reports were in English or a language for which a translator was available. Computerized searches were conducted and all potentially relevant titles and abstracts were extracted. Retrieved articles were assessed for relevance independently by two reviewers, based on predetermined criteria. Articles that met all criteria for relevance were assessed for methodological quality based on predetermined criteria. Articles judged to have the appropriate quality by both reviewers were included in the analysis. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the two authors. Meta-analyses were conducted for each intervention where the same outcome measures and/or instruments were used within comparable time points. MAIN RESULTS: This review detected 31 eligible randomized controlled trials involving four major groups of developmental care interventions, 19 sub-groups and multiple clinical outcomes. The results of the review indicate that developmental care interventions demonstrate some benefit to preterm infants with respect to: improved short-term growth outcomes, decreased respiratory support, decreased length and cost of hospital stay, and improved neurodevelopmental outcomes to 24 months corrected age. These findings were based on two or three small trials for each outcome, and did not involve meta-analyses of more than two trials for any one outcome. Although a number of other benefits were demonstrated, those results were from single studies with small sample sizes. The lack of blinding of the assessors was a significant methodological flaw in half of the studies. The cost of the interventions and personnel was not considered in any of the studies. REVIEWER'S CONCLUSIONS: Because of the inclusion of multiple interventions in most studies, the determination of the effect of any single intervention is difficult. Although there is evidence of some benefit of developmental care interventions overall, and no major harmful effects reported, there were a large number of outcomes for which no or conflicting effects were demonstrated. The single trials that did show a significant effect of an intervention on a major clinical outcome were based on small sample sizes, and the findings were often not supported in other small trials. Before a clear direction for practice can be supported, evidence demonstrating more consistent effects of developmental care interventions on important short- and long-term clinical outcomes is needed. The economic impact of the implementation and maintenance of developmental care practices should be considered by individual institutions. PMID- 11034731 TI - Nutritional supplementation for hip fracture aftercare in the elderly. AB - BACKGROUND: Fractures of the hip are an important cause of later ill health and mortality in elderly people. People with hip fractures are often malnourished at the time of fracture, and have poor food intake in hospital. OBJECTIVES: This review assesses the effects of nutritional interventions in elderly people recovering from hip fracture. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, the Cochrane Controlled Trials Register, Medline, Nutrition Abstracts and Reviews, Embase, Biosis, Cinahl, Healthstar and reference lists. We contacted investigators, and hand searched The American Journal of Clinical Nutrition, Proceedings of the Nutrition Society, Clinical Nutrition and The Journal of Parenteral and Enteral Nutrition. Date of the most recent search: January 2000. SELECTION CRITERIA: Randomised and quasi randomised trials of nutritional interventions of mainly older patients (aged over 65 years) with hip fracture. DATA COLLECTION AND ANALYSIS: Trial allocation to included, excluded and awaiting assessment categories, was by consensus. Both reviewers independently extracted data and assessed trial quality. Additional information was sought from all trialists. Pooling of data for primary outcomes and select exploratory analyses were undertaken. MAIN RESULTS: Fifteen randomised trials involving 1054 participants were included. Overall the quality of trials was poor; specifically in terms of allocation concealment, assessor blinding and intention to treat analysis. This, and the limited availability of outcome data, mean that the following results must be interpreted with caution. Oral multinutrient feeds (providing non-protein energy, protein, some vitamins and minerals), evaluated by five trials, may reduce unfavourable outcome (death or complications) (14/66 versus 26/73; relative risk 0.52, 95% confidence interval 0.32 to 0.84), but did not demonstrate an effect on mortality (12/91 versus 14/97; relative risk 0.85, 95% confidence interval 0.42 to 1.70). Four trials, examining nasogastric multinutrient feeding, showed no evidence for an effect on mortality (relative risk 0.99, 95% confidence interval 0.50 to 1.97), but the studies were heterogeneous regarding case-mix. Insufficient information was provided to evaluate unfavourable outcome. The effect of protein in an oral feed, tested in three trials, showed no evidence for an effect on mortality (relative risk 1.38, 95% confidence interval 0.82 to 2.34). It may have reduced the number of long term complications and days spent in rehabilitation wards. Two trials, testing intravenous thiamin (vitamin B1) and other water soluble vitamins, or 1 alpha-hydroxycholecalciferol (an active form of vitamin D) respectively, produced no evidence of benefit for either vitamin supplement. REVIEWER'S CONCLUSIONS: The strongest evidence for the effectiveness of nutritional supplementation exists for oral protein and energy feeds, but the evidence is still very weak. Future trials are required which overcome the defects of the reviewed studies, particularly inadequate size, methodology and outcome assessment. PMID- 11034732 TI - Epidural local anaesthetics versus opioid-based analgesic regimens on postoperative gastrointestinal paralysis, PONV and pain after abdominal surgery. AB - BACKGROUND: Gastrointestinal paralysis, nausea and vomiting, and pain, are major clinical problems following abdominal surgery. Anaesthetic and analgesic techniques that reduce pain and postoperative nausea and vomiting (PONV), and prevent or reduce postoperative ileus, may reduce postoperative morbidity, duration of hospitalisation and hospital costs. OBJECTIVES: To compare effects of postoperative epidural local anaesthetic with regimens based on systemic or epidural opioids, on postoperative gastrointestinal function, postoperative pain, PONV and surgical/anaesthetic complications. SEARCH STRATEGY: Trials were identified by computerised searches of the Cochrane Controlled Trials Register, MEDLINE, EMBASE and by checking the reference lists of trials and review articles. SELECTION CRITERIA: Randomised controlled trials comparing the effects of postoperative epidural local anaesthetic with systemic or epidural opioids. DATA COLLECTION AND ANALYSIS: Collected data included treatment in active (local anaesthetic) and control (opioid based) groups, time to first postoperative stool, time to first postoperative flatus, gastric emptying measured by the paracetamol absorption test, duration of the passage of barium sulphate, pain assessments, use of supplementary analgesics, nausea, vomiting and surgical/anaesthetic complications. MAIN RESULTS: Most studies in this review involved a small number of patients. Furthermore half of the studies indicated a poor level of methodology in particular regarding blinding and report of withdrawals. Heterogeneity of included studies was substantial. Results consistently showed reduced time to return of gastrointestinal function in the epidural local anaesthetic group compared with groups receiving systemic or epidural opioid (37 hours and 24 hours, respectively). Postoperative pain was comparable. Two studies compared the effect of epidural local anaesthetic with a combination of epidural local anaesthetic and opioid on gastrointestinal function. One study favoured epidural local anaesthetic and one study was indifferent. A meta analysis of five of eight studies comparing the effect of epidural local anaesthetic with a combination of epidural local anaesthetic and opioid on postoperative pain, yielded a reduction in VAS pain scores (0-100 mm) on the first postoperative day of 15 mm, in favour of the combination. No significant differences in PONV were observed between epidural local anaesthetic and opioid based regimens. REVIEWER'S CONCLUSIONS: Administration of epidural local anaesthetics to patients undergoing laparotomy reduce gastrointestinal paralysis compared with systemic or epidural opioids, with comparable postoperative pain relief. Addition of opioid to epidural local anaesthetic may provide superior postoperative analgesia compared with epidural local anaesthetics alone. The effect of additional epidural opioid on gastrointestinal function is so far unsettled. Randomized, controlled trials comparing the effect of combinations of epidural local anaesthetic and opioid with epidural local anaesthetic alone on postoperative gastrointestinal function and pain are warranted. PMID- 11034733 TI - Home intravenous antibiotics for cystic fibrosis. AB - BACKGROUND: Recurrent endobronchial infection in cystic fibrosis requires treatment with intravenous antibiotics for several weeks, which is usually administered in hospital, affecting health costs and quality of life for patients and their families. It is not known whether patients receiving intravenous treatment at home have better or equivalent health outcomes, if costs are reduced or if it is preferred than in-hospital treatment. Home treatment requires training to patients and carers and usually needs a few previous days in hospital. OBJECTIVES: To determine whether home intravenous antibiotic therapy in cystic fibrosis is as effective as in-patient intravenous antibiotic therapy and if it is preferred by patients and/or families. SEARCH STRATEGY: References to trials were obtained from the specialist cystic fibrosis trials register held by the editorial base of the Cochrane Cystic Fibrosis and Genetic Disorders Group. Handsearching of the abstracts books of all Spanish Conferences on cystic fibrosis and the last European Conference (Stockholm, 2000) was carried out by authors. SELECTION CRITERIA: Randomised controlled trials where home intravenous antibiotic treatment for patients with cystic fibrosis was compared with in hospital intravenous antibiotic treatment, including adults and children with cystic fibrosis. All kinds of antibiotics and regimens administered intravenous were included. DATA COLLECTION AND ANALYSIS: Three reviewers independently selected the trials to be included in the review, assessed methodological quality of each trial and extracted data using a standardised form. Because of several limitations, narrative synthesis was used at this stage. MAIN RESULTS: One study was included with 17 patients aged 10 to 41 years with an infective exacerbation by Pseudomonas aeruginosa. All their 31 admissions were analysed as independent events. Outcomes were measured at 21 days of follow-up after initiation of treatment. Home patients had fewer investigations performed than hospital patients (p<0.002) and general activity was higher in the home group. No differences were found for clinical outcomes, adverse events, complications of intravenous lines or line changes or time to next admission. Home patients received less low-dose home maintenance antibiotic. Quality of life measures showed no differences for dyspnoea and emotional state, but fatigue and mastery were worse for home patients, possibly due to a higher general activity and need of support. Personal, family, sleeping and eating disruptions were less important for home than hospital admissions. Home therapy was cheaper for families and the hospital. Indirect costs were not determined. REVIEWER'S CONCLUSIONS: The current evidence is restricted to one small study. It suggests that in the short term home therapy does not harm patients and in general reduces social disruptions. The decision to attempt home treatment should be based on an individual basis and appropriate local resources. More research is urgently required. PMID- 11034734 TI - Massage for low back pain. AB - BACKGROUND: Low back pain is one of the most common and costly musculoskeletal problems in modern societies. Proponents of massage therapy claim it can minimize pain and disability, and speed return to normal function. OBJECTIVES: To assess the effects of massage therapy for non-specific low back pain. SEARCH STRATEGY: We searched Medline, Embase, Cochrane Controlled Trials Register, Healthstar, CINAHL and Dissertation abstracts from 1966 to 1999 with no language restrictions. References in the included studies and in reviews of the literature were also screened. Contact with content experts and massage associations were also made. SELECTION CRITERIA: This review included randomized, quasi-randomized or controlled clinical trials that investigated the use of any type of massage (using the hands or a mechanical device) as a treatment for nonspecific low back pain. DATA COLLECTION AND ANALYSIS: One reviewer applied the selection criteria and extracted the data. Two reviewers (one blinded to authors, institutions and journals) independently assessed the quality of each trial. A qualitative analysis (best-evidence synthesis) was performed due to clinical heterogeneity among the included trials and insufficient data reported. MAIN RESULTS: Four randomized controlled trials met the inclusion criteria. Two trials were of high and two of low methodological quality. None evaluated massage as the main intervention. Rather, it was the control intervention in studies evaluating manipulation, electrical stimulation, and a lumbar corset. There is limited evidence showing that massage is less effective than manipulation immediately after the first session and moderate evidence showing it is less effective than TENS during the course of sessions in relieving pain and improving activity. At the completion of treatment and at 3 weeks after discharge there is no difference among massage and manipulation, electrical stimulation or corsets, but this evidence is limited. REVIEWER'S CONCLUSIONS: Based on the studies reviewed, there is insufficient evidence to recommend massage as a stand-alone treatment for non specific low back pain. There is a need for high quality controlled trials to further evaluate the effects of massage for this condition. PMID- 11034735 TI - Subjective barriers to prevent wandering of cognitively impaired people. AB - BACKGROUND: People with dementia often wander, at times putting themselves at risk and presenting challenges to carers and institutional staff. Traditional interventions to prevent wandering include restraint, drugs and locked doors. Cognitively impaired people may respond to environmental stimuli (sounds, images, smells) in ways distinct from healthy people. This has led to trials of visual and other selective barriers (such as mirrors, camouflage, grids/stripes of tape) that may reduce wandering. OBJECTIVES: We assess the effect of subjective exit modifications on the wandering behaviour of cognitively impaired people. The second objective is to inform the direction and methods of future research. SEARCH STRATEGY: The search strategy includes electronic searches of relevant bibliographic and trials databases, citation indices and relevant medical journals. SELECTION CRITERIA: Randomized controlled trials and controlled trials provide the highest quality evidence, but interrupted time series are also considered as they may contribute useful information. Participants are people with dementia or cognitive impairment who wander, of any age, and in any care environment - hospital, other institution, or their own home. Interventions comprise exit modifications that aim to function as subjective barriers to prevent the wandering of cognitively impaired people. Locks, physical restraints, electronic tagging and other types of barrier are not included. DATA COLLECTION AND ANALYSIS: The criteria for inclusion or exclusion of studies are applied independently by two reviewers. All outcomes that are meaningful to people making decisions about the care of wanderers are recorded. These include the number of exits or carer interventions, resource use, acceptability of the intervention and the effects on carer and wanderer anxiety or distress. heterogeneity of clinical area, of study design and of intervention was substantial. MAIN RESULTS: No randomized controlled or controlled trials were found. The other experimental studies that we identified were unsatisfactory. Most were vulnerable to bias, particularly performance bias; most did not classify patients according to type or severity of dementia; in all studies, outcomes were measured only in terms of wandering frequency rather than more broadly in terms of quality of life, resource use, anxiety and distress; no studies included patients with delirium; no studies were based in patients' homes. REVIEWER'S CONCLUSIONS: There is no evidence that subjective barriers prevent wandering in cognitively impaired people. PMID- 11034736 TI - Oral or topical nasal steroids for hearing loss associated with otitis media with effusion in children. AB - BACKGROUND: OME is common and may cause hearing loss with associated developmental delay. Treatment remains controversial. The effect of both systemic and intra-nasal steroids on effusions has been assessed by randomised controlled trials. OBJECTIVES: To examine evidence for or against treating children with hearing loss associated with OME with systemic or topical nasal steroids. SEARCH STRATEGY: Searches were conducted in February 2000. We searched the Cochrane Controlled Trials Register using the terms 'otitis-media', 'otitis media with effusion', 'glue ear', or 'OME', and 'steroids', 'glucocorticoids, synthetic', 'glucocorticoids, topical', 'anti-inflammatory agents, steroidal'. EMBASE and MEDLINE were also searched for additional information. SELECTION CRITERIA: Randomised controlled trials of oral and topical nasal steroids, either alone or in combination with another agent such as an antibiotic, were included. EXCLUSIONS: publications in abstract form only since adequate appraisal was not possible; uncontrolled, non-randomised or retrospective studies; studies reporting outcomes with ears (rather than children) as the unit of analysis. DATA COLLECTION AND ANALYSIS: Data were extracted from the published reports by the two authors independently (CCB and JH van der V) using standardised data extraction forms and methodology. The methodological quality of the included studies were independently assessed by the two authors using the scheme described in the Cochrane Handbook. Dichotomous results were expressed as an odds ratio using a fixed effects model together with the 95% confidence intervals. Continuous data were analysed using the weighted mean difference in a fixed effects model. Tests for heterogeneity between studies were performed using a Mantel-Haenszel approach. In trials with a cross over design, post-crossover treatment data were not used. MAIN RESULTS: No study prospectively documented hearing loss associated with OME prior to randomisation. Follow up was short term. No serious or lasting side effects were reported in the four studies that did mention side effects. Most comparisons involved small numbers of subjects. The odds ratio for OME persisting after short term follow up for children treated with oral steroids plus antibiotic compared to control plus antibiotic was 0.32 (95% CI 0.20 to 0.52). However there was significant heterogeneity between studies (p<0.01). Trends favoured steroids for most other comparisons, but confidence intervals included unity. There was no evidence of benefit for steroid treatment in the longer term, and no study assessed effect of steroid treatment on language development. REVIEWER'S CONCLUSIONS: There is evidence that steroids combined with an antibiotic lead to a quicker resolution of OME in the short term. However, there is not evidence for long term benefit from treating hearing loss associated with OME with either oral or topical nasal steroids. These treatments are therefore not recommended at the present time. Future studies should document hearing loss associated with OME before the start of study treatment. Follow up should be longer and ideally include symptom, audiometry and developmental outcomes. Data should not be presented with ears as the unit of analysis. PMID- 11034737 TI - Ziprasidone for schizophrenia and severe mental illness. AB - BACKGROUND: Typical antipsychotic drugs are widely used as the first line treatment for people with schizophrenia. However, the atypical class of antipsychotic drugs are making important inroads into this approach. Atypical is a widely used term used to describe some antipsychotics which have a low propensity to produce movement disorders and raise serum prolactin. There is some suggestion that the different adverse effect profiles of atypical antipsychotic group make them more acceptable to people with schizophrenia. Ziprasidone is one of the newer atypicals with a high serotonin receptor affinity. OBJECTIVES: To determine the effects of ziprasidone compared with placebo, typical and other atypical antipsychotic drugs for schizophrenia and related psychoses. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1980-1999), The Cochrane Library (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (January 1999), CINAHL (1982-1999), EMBASE (1980-1999), MEDLINE (1966-1999), LILACS (1982 1996), PSYNDEX (1977-1999) and PsycLIT (1974-1999) were undertaken. In addition, pharmaceutical databases on the Dialog Corporation Datastar and Dialog services were searched. References of all identified studies were searched for further trials. Pharmaceutical companies (Pfizer - the manufacturer of ziprasidone - and the manufacturers of all comparator drugs) and first authors of all included trials were contacted. SELECTION CRITERIA: All randomised controlled trials that compared ziprasidone to other treatments for schizophrenia and schizophrenia-like psychoses were included by independent assessment. DATA COLLECTION AND ANALYSIS: Citations and, where possible, abstracts were independently inspected by reviewers, papers ordered, re-inspected and quality assessed. Data were independently extracted. Data were excluded if loss to follow up was greater than 50%. For homogeneous dichotomous data the risk ratio (RR), 95% confidence interval (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to-treat basis. For continuous data, weighted mean differences were calculated (WMD). All data were inspected for heterogeneity. MAIN RESULTS: Data for this compound range from very short (one week) studies of the intramuscular preparation, to trials lasting over six months. For measures of mental state ziprasidone seems more effective than placebo (RR 0.8 CI 0.7-0.9) and as effective as haloperidol (RR 0.8 CI 0.7-1). It is less likely than haloperidol to cause movement disorders (RR 0.4 CI 0.2-0.6), but causes more nausea and vomiting (RR 2.1 CI 1.2-3.8). The injected form of the drug causes more pain at the injection site than haloperidol (RR 5.3 CI 1.3-22). REVIEWER'S CONCLUSIONS: Currently data are limited. Ziprasidone may be an effective antipsychotic with less extrapyramidal effects than haloperidol. It also, however, causes more nausea and vomiting than the typical drugs, and, at present, there is no data suggesting that it is different to other atypical compounds. Well planned, conducted and reported long term randomised trials are needed if ziprasidone is to be accepted into everyday use. PMID- 11034738 TI - Intravesical Bacillus Calmette-Guerin in Ta and T1 Bladder Cancer. AB - BACKGROUND: Intravesical therapy with Bacillus Calmette-Guerin (BCG) aims to reduce the incidence of tumour recurrence following transurethral resection (TUR) for patients with superficial bladder cancer. OBJECTIVES: The objective of this review was to compare the incidence of tumour recurrence after the standard therapy of transurethral resection versus transurethral resection plus intravesical Bacillus Calmette-Guerin. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (March 2000), Medline (February, 2000), EMBASE (February, 2000), Cancerlit (February, 2000), Healthstar (February, 2000), Database of Abstracts of Reviews of Effectiveness (February, 2000) and the Bath Information Data Service. The Proceedings of the American Society Clinical Oncology was hand searched (1996 - 1999). SELECTION CRITERIA: Randomised or quasi randomised trials of transurethral resection alone versus transurethral resection plus intravesical Bacillus Calmette-Guerin. Patients with Ta and T1 bladder cancer of medium or high risk of tumour recurrence, were eligible for inclusion. DATA COLLECTION AND ANALYSIS: Four reviewers assessed trial quality and two abstracted the data independently. The Peto odds ratios and log hazard ratios were determined to compare the number of patients with disease recurrence at 12 months and the rate of recurrence, respectively. MAIN RESULTS: Six randomised trials were included involving 585 eligible patients. There were significantly fewer patients with disease recurrence at 12 months in the BCG plus TUR group compared to those that received TUR alone (odds ratio 0.30, CI 0.21, 0.43). The overall log hazard ratio for recurrence (-0.83, variance 0.02) indicated a significant benefit of BCG treatment in reducing tumour recurrence. Toxicities associated with BCG consisted mainly of cystitis (67%), haematuria (23%), fever (25%) and urinary frequency (71%). No BCG-induced deaths were reported. REVIEWER'S CONCLUSIONS: In patients with medium/high risk Ta or T1 bladder cancer, immunotherapy with intravesical BCG following TUR appears to provide a significant advantage over TUR alone in delaying tumour recurrence. PMID- 11034739 TI - Vitamin K antagonists or low-molecular-weight heparin for the long term treatment of symptomatic venous thromboembolism. AB - BACKGROUND: Patients who have had an episode of symptomatic venous thromboembolism are usually treated for at least five days with intravenous unfractionated heparin or subcutaneous low-molecular-weight heparin. Thereafter, they received a three month course of a vitamin K antagonist, with a dose adjusted to achieve an International Normalized Ratio between 2.0 and 3.0. Some patients have contraindications to vitamin K antagonists. In addition, treatment with vitamin K antagonists has the disadvantage of regular laboratory measurements. OBJECTIVES: The objective of this review was to evaluate the efficacy and safety of long-term treatment of symptomatic venous thromboembolism with low-molecular-weight heparins compared with vitamin K antagonists. SEARCH STRATEGY: Computerized searches of MEDLINE, EMBASE and Current Contents were made and relevant journals were hand-searched using the search strategy described by the Cochrane Peripheral Vascular Disease Group. In addition, randomized clinical trials were located through personal communication with colleagues. Where necessary, the reviewers contacted pharmaceutical companies for further information. SELECTION CRITERIA: Two reviewers evaluated studies independently for methodological quality. DATA COLLECTION AND ANALYSIS: Two reviewers reviewed and extracted data independently using a standard form. Primary analysis concerned all patients in the studies during the period of randomized treatment. Additional separate analyses were performed for category I and category II studies; studies that used similar initial treatments in both study arms and those that used different treatment regimes during the initial treatment; and the total period of follow-up in the different studies. MAIN RESULTS: Five studies were identified that fulfilled our predefined criteria (three category I and two category II studies). When all five studies were combined, a statistically non significant reduction of the risk of recurrent symptomatic venous thromboembolism in favor of low-molecular-weight heparin treatment (OR 0.72; 95% CI [0.42, 1.23]) was found. In category I studies, analysis of the pooled data showed a statistically non-significant reduction of the risk of recurrent symptomatic venous thromboembolism in favor of low-molecular-weight heparin treatment (OR 0.75; 95% CI [0.40, 1.39]). This OR was mainly due to one possibly confounded study, and after omitting this study from the analysis a statistically non significant reduction of the risk of recurrent symptomatic venous thromboembolism in favor of vitamin K antagonist treatment remained (OR 1.95; 95% CI [0.74, 5.19]). No differences in the risk of bleeding (OR 0.63; 95% CI [0.21, 1.88]) and mortality (OR 1.13; 95% CI [0.47, 2.69] were observed. REVIEWER'S CONCLUSIONS: Low-molecular-weight heparins are possibly as effective and safe as vitamin K antagonists in the prevention of recurrent symptomatic venous thromboembolism after an episode of symptomatic deep venous thrombosis, but have the disadvantage of much higher medicinal costs. Treatment with low-molecular-weight heparin is possibly a safe alternative in some patients; for example patients who live in geographically inaccessible places; patients who are reluctant to go to the thrombosis service on a regular basis; and patients with contraindications to vitamin K antagonists (e.g. pregnant women). Therefore, in the absence of definitive evidence on the safety and efficacy of low-molecular-weight heparins compared with vitamin K antagonists, we believe that treatment with vitamin K antagonists is still the treatment of choice in the prevention of recurrent symptomatic venous thromboembolism after an episode of deep venous thrombosis, in the majority of patients. PMID- 11034740 TI - Once daily versus multiple daily dosing with intravenous aminoglycosides for cystic fibrosis. AB - BACKGROUND: Patients with cystic fibrosis, who are chronically colonised with the organism Pseudomonas aeruginosa, often require repeated courses of intravenous aminoglycoside antibiotics for the management of pulmonary exacerbations. The properties of aminoglycosides suggest that they could be given in higher concentrations less often. OBJECTIVES: To assess the effectiveness and safety of once-daily versus multiple-daily dosing of intravenous aminoglycoside antibiotics for the management of pulmonary exacerbations in cystic fibrosis. SEARCH STRATEGY: We searched the Cystic Fibrosis specialist trials register held at the Cochrane Cystic Fibrosis and Genetic Disorders Group's editorial base, which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Date of the most recent search: February 2000. SELECTION CRITERIA: All randomised controlled trials, whether published or unpublished, in which once daily dosing of aminoglycosides has been compared with multiple-daily dosing in terms of efficacy and/or toxicity, in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: Both reviewers independently extracted data and assessed trial quality. Authors of one study were contacted to obtain missing information. MAIN RESULTS: Two trials reporting results from a total of 70 patients were included in this review. Both trials compared once-daily dosing with thrice-daily dosing reporting data on Forced Expiratory Volume at one second (FEV1), Forced Vital Capacity (FVC), nutritional status and side effects. There was no significant difference in efficacy or in the incidence of ototoxicity and nephrotoxicity between treatment groups. REVIEWER'S CONCLUSIONS: Despite a lack of difference between the two groups we feel that these results should be viewed with caution as the numbers of patients involved was small and lacks the power to detect a difference between the groups. This systematic review has highlighted the need for a well designed, adequately-powered, multicentre, randomised controlled trial assessing the efficacy of once-daily versus multiple-daily dosing of intravenous aminoglycosides for pulmonary exacerbations in cystic fibrosis. PMID- 11034741 TI - Parent-training programmes for improving maternal psychosocial health. AB - BACKGROUND: The prevalence of mental health problems in women is 1:3 and such problems tend to be persistent. There is evidence from a range of studies to suggest that a number of factors relating to maternal psychosocial health can have a significant effect on the mother-infant relationship, and that this can have consequences for the psychological health of the child. It is now thought that parenting programmes may have an important role to play in the improvement of maternal psychosocial health. OBJECTIVES: The objective of this review is to address whether group-based parenting programmes are effective in improving maternal psychosocial health including anxiety, depression and self-esteem. SEARCH STRATEGY: A range of biomedical, social science, educational and general reference electronic databases were searched including MEDLINE, EMBASE CINAHL, PsychLIT, ERIC, ASSIA, Sociofile and the Social Science Citation Index. Other sources of information included the Cochrane Library (SPECTR, CENTRAL), and the National Research Register (NRR). SELECTION CRITERIA: Only randomised controlled trials were included in which participants had been randomly allocated to an experimental and a control group, the latter being either a waiting-list, no treatment or a placebo control group. Studies had to include at least one group based parenting programme, and one standardised instrument measuring maternal psychosocial health. DATA COLLECTION AND ANALYSIS: A systematic critical appraisal of all included studies was undertaken using the Journal of the American Medical Association (JAMA) published criteria. The data were summarised using effect sizes but were not combined in a meta-analysis due to the small number of studies within each group and the presence of significant heterogeneity. MAIN RESULTS: A total of 22 studies were included in the review but only 17 provided sufficient data to calculate effect sizes. These 17 studies reported on a total of 59 outcomes including depression, anxiety, stress, self esteem, social competence, social support, guilt, mood, automatic thoughts, dyadic adjustment, psychiatric morbidity, irrationality, anger and aggression, mood, attitude, personality, and beliefs. Approximately 22% of the outcomes measured suggested significant differences favouring the intervention group. A further 40% showed differences favouring the intervention group but which failed to achieve conventional levels of statistical significance, in some cases due to the small numbers that were used. Approximately 38% of outcomes suggested no evidence of effectiveness. REVIEWER'S CONCLUSIONS: It is suggested that parenting programmes can make a significant contribution to the improvement of psychosocial health in mothers. While the critical appraisal suggests some variability in the quality of the included studies, it is concluded that there is sufficient evidence to support their use with diverse groups of parents. However, it is also suggested that some caution should be exercised before the results are generalised to parents irrespective of the level of pathology present, and that further research is still required. PMID- 11034742 TI - Hypertonic versus isotonic crystalloid for fluid resuscitation in critically ill patients. AB - BACKGROUND: Hypertonic solutions are considered to have a greater ability to expand blood volume and thus elevate blood pressure and can be administered as a small volume infusion over a short time period. On the other hand, the use of hypertonic solutions for volume replacement may also have important disadvantages. OBJECTIVES: To determine whether hypertonic crystalloid decreases mortality in patients with hypovolaemia with and without head injuries. SEARCH STRATEGY: We searched MEDLINE, EMBASE, The Cochrane Controlled Trials Register and the Specialised register of the Injuries Group. We checked reference lists of all articles identified and searched the National Research Register. SELECTION CRITERIA: Randomised trials comparing hypertonic to isotonic crystalloid in patients with trauma, burns or undergoing surgery. DATA COLLECTION AND ANALYSIS: Two authors independently extracted the data and assessed the quality of the trials. MAIN RESULTS: Sixteen trials were identified with a total of 837 participants. Data on death were obtained in twelve of the studies. Only one trial reported data on disability. The pooled RR for death in trauma patients was 0.84 (95% CI 0.61-1.16), and in patients with burns 1.49 (95% CI 0.56-3.95), and in patients undergoing surgery 0.62 (95% cI 0.08-4.57). In the one trial that gave data on disability using the Glasgow Outcome Scale the relative risk was 0.99 (95% CI 0.06-15.93). REVIEWER'S CONCLUSIONS: This review does not give us enough data to be able to say whether hypertonic crystalloid is better than isotonic crystalloid for the resuscitation of patients with trauma, burns, or those undergoing surgery. However, the confidence intervals are wide and do not exclude clinically significant differences. Further trials are needed comparing hypertonic to isotonic crystalloid. Trials need to be large enough to detect a clinically important difference. PMID- 11034743 TI - Feather vs. non-feather bedding for asthma. AB - BACKGROUND: Two recent epidemiological studies have reported that children using non-feather pillows suffered from more frequent episodes of wheeze than those using feather pillows OBJECTIVES: To evaluate the efficacy of using feather bedding in the control of asthma symptoms. SEARCH STRATEGY: The Cochrane Airways Group asthma clinical trials register, derived from MEDLINE, EMBASE and hand searching of major journals, was searched using the terms: feather OR bed* OR linen* OR pillow SELECTION CRITERIA: Only randomised or controlled clinical trials were to be included. DATA COLLECTION AND ANALYSIS: No trials met the inclusion criteria for the review. MAIN RESULTS: 126 abstracts were identified. Ten of these were identified as possibly meeting the entry criteria, but on review of the full paper not were suitable. The reasons for exclusion were: not a randomised trial (n=6); allocation of bedding type combined with another intervention (n=4). REVIEWER'S CONCLUSIONS: Whilst recent epidemiological studies suggest that feather bedding is associated with less frequent wheeze than man made fibre fillings, the evidence currently available is insufficient to assess the clinical benefits of feather bedding in the management of asthma. PMID- 11034744 TI - Leukotriene receptor antagonists for non-cystic fibrosis bronchiectasis. AB - BACKGROUND: Leukotriene receptor antagonists are a new class of drug that were initially identified for use in asthma. As they have an effect on neutrophil mediated inflammation, they may be of benefit in bronchiectasis. OBJECTIVES: To determine whether leukotriene receptor antagonists have any additive benefit over and above conventional treatment for bronchiectasis (usually consisting of antibiotics and postural drainage). SEARCH STRATEGY: The Cochrane Airways Group clinical trials register derived from MEDLINE, EMBASE and hand searching of major journals was searched using the terms:Bronchiec* AND leukotrien* OR anti-leuk* OR cysteinyl, Bronchiec* AND monteluk*, Bronchiec* AND zafirluk* SELECTION CRITERIA: Only randomised, controlled trials were considered DATA COLLECTION AND ANALYSIS: The results of searches were analysed by both authors MAIN RESULTS: No randomised, controlled trials were identified REVIEWER'S CONCLUSIONS: Further research is required to establish any benefit from the use of leukotriene antagonists in bronchiectasis. PMID- 11034745 TI - Surgery vs non-surgical treatment for bronchiectasis. AB - BACKGROUND: Standard treatment for bronchiectasis comprises postural drainage and various regimes of antibiotic therapy. If the disease is confined to localised areas of lung, surgical resection of the affected segments is often performed. OBJECTIVES: To assess the benefit of surgical resection compared with standard ("conservative") treatment. SEARCH STRATEGY: The Cochrane Airways Group trials register derived from MEDLINE, EMBASE and hand searching of major journals was searched using the terms [bronchiect* AND surg* OR resection OR lobect* OR pneumonect* OR segementect*]. SELECTION CRITERIA: Only randomised, controlled trials were considered DATA COLLECTION AND ANALYSIS: The titles, abstracts and citations were independently reviewed by the two reviewers to assess potential relevance for full review. STATISTICAL CONSIDERATIONS Not applicable MAIN RESULTS: No randomised or controlled clinical trials were found, other than case series or case-controlled studies. REVIEWER'S CONCLUSIONS: Surgical treatment of bronchiectasis is widely used, but there appear to be no randomised controlled trials. It is not possible to provide an unbiased estimate of its benefit compared to conservative therapy. PMID- 11034746 TI - Antifungal therapies for allergic bronchopulmonary aspergillosis in people with cystic fibrosis. AB - BACKGROUND: Allergic Bronchopulmonary Aspergillosis (ABPA) is an allergic reaction to colonisation of the lungs with the fungus Aspergillus fumigatus and affects around 10% people with cystic fibrosis. ABPA is associated with an accelerated decline in lung function. Corticosteroids, in high doses, are the main treatment for ABPA although the long-term benefits are not clear and their many side effects are well documented. A group of compounds, the azoles, have activity against Aspergillus fumigatus and have been proposed as an alternative treatment for ABPA. Of this group, Itraconazole is the most active. A separate antifungal compound, Amphotericin B has been employed in aerosolised form to treat invasive infection with Aspergillus fumigatus, and may have potential for the treatment of ABPA. Antifungal therapy for ABPA in cystic fibrosis needs to be evaluated. OBJECTIVES: The review tested the hypotheses that antifungal interventions for the treatment of ABPA in cystic fibrosis: 1. improve clinical status compared to placebo or standard therapy (no placebo); 2. do not have unacceptable adverse effects. If benefit was demonstrated, the optimal type, duration and dose of antifungal therapy was assessed. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group specialist trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. In addition, pharmaceutical companies were approached. SELECTION CRITERIA: Randomised controlled trials, published or unpublished, where antifungal treatments have been compared to either placebo or no treatment, or where different doses of the same treatment have been used in the treatment of ABPA in patients with cystic fibrosis. DATA COLLECTION AND ANALYSIS: No completed randomised controlled trials were identified. MAIN RESULTS: No completed randomised controlled trials were identified. REVIEWER'S CONCLUSIONS: At present, there are no randomised controlled trials to evaluate the use of antifungal therapies for the treatment of ABPA in people with cystic fibrosis. Trials with clear outcome measures are needed to properly evaluate this potentially useful treatment for cystic fibrosis. PMID- 11034747 TI - Continuous distending pressure for respiratory distress syndrome in preterm infants. AB - BACKGROUND: Respiratory distress syndrome (RDS) is the single most important cause of morbidity and mortality in preterm infants (Greenough 1998, Bancalari 1992). Intermittent positive pressure ventilation (IPPV) with surfactant is the standard treatment for the condition. The major difficulty with IPPV is that it is invasive, resulting in airway and lung injury and contributing to the development of chronic lung disease. OBJECTIVES: In spontaneously breathing preterm infants with RDS, to determine if continuous distending pressure (CDP) reduces the need for IPPV and associated morbidity without adverse effects. SEARCH STRATEGY: The standard search strategy of the Neonatal Review group was used. This included searches of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register, MEDLINE (1966-Jan. 2000), previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants, journal hand searching mainly in the English language. SELECTION CRITERIA: All trials using random or quasi-random patient allocation of newborn infants with RDS were eligible. Interventions were continuous distending pressure including continuous positive airway pressure (CPAP) by mask, nasal prong, nasopharyngeal tube, or endotracheal tube, or continuous negative pressure (CNP) via a chamber enclosing the thorax and lower body, compared with standard care. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Collaboration and its Neonatal Review Group, including independent assessment of trial quality and extraction of data by each author, were used. MAIN RESULTS: CDP is associated with a lower rate of failed treatment (death or use of assisted ventilation), overall mortality, and mortality in infants with birthweights above 1500 g. The use of CDP is associated with an increased rate of pneumothorax. REVIEWER'S CONCLUSIONS: In preterm infants with RDS the application of CDP either as CPAP or CNP is associated with some benefits in terms of reduced respiratory failure and reduced mortality. CDP is associated with an increased rate of pneumothorax. The applicability of these results to current practice is difficult to assess, given the outdated methods to administer CDP, low use of antenatal corticosteroids, non availability of surfactant and the intensive care setting of the 1970s when these trials were done. Where resources are limited, such as in developing countries, CPAP for RDS may have a clinical role. Further research is required to determine the best mode of administration and its role in modern intensive care settings PMID- 11034748 TI - Prevention of NSAID-induced gastroduodenal ulcers. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) are important agents in the management of arthritic and inflammatory conditions, and are among the most frequently prescribed medications in North America and Europe. However, there is overwhelming evidence linking these agents to a variety of gastrointestinal (GI) toxicities. OBJECTIVES: To review the effectiveness of common interventions for the prevention of NSAID induced upper GI toxicity. SEARCH STRATEGY: A literature search was conducted, according to the Cochrane methodology for identification of randomized controlled trials in electronic databases, including MEDLINE from 1966 to January 2000, Current Contents for 6 months prior to January 2000, Embase to Febuary 1999, and a search of the Cochrane Controlled Trials Register from 1973 to 1999. Recent conference proceedings were reviewed and content experts and companies were contacted. SELECTION CRITERIA: Randomized controlled clinical trials (RCTs) of prostaglandin analogues (PA), H2-receptor antagonists (H2RA) or proton pump inhibitors (PPI) for the prevention of chronic NSAID induced upper GI toxicity were included. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted data regarding population characteristics, study design, methodological quality and number of patients with endoscopic ulcers, ulcer complications, symptoms, overall drop outs, drop outs due to symptoms. Dichotomous data was pooled using Revman V3.1. Heterogeneity was evaluated using a chi square test. MAIN RESULTS: Thirty-three RCTs met the inclusion criteria. All doses of misoprostol significantly reduced the risk of endoscopic ulcers. Misoprostol 800 ug/day was superior to 400 ug/day for the prevention of endoscopic gastric ulcers (RR=0.18, and RR=0.38 respectively, p=0.0055). A dose response relationship was not seen with duodenal ulcers. Misoprostol caused diarrhea at all doses, although significantly more at 800ug/day than 400ug/day (p=0.0012). Misoprostol was the only prophylactic agent documented to reduce ulcer complications. Standard doses of H2RAs were effective at reducing the risk of endoscopic duodenal (RR=0.24; 95% CI: 0.10-0.57) but not gastric ulcers(RR=0.73; 95% CI:0.50-1.09). Both double dose H2RAs and PPIs were effective at reducing the risk of endoscopic duodenal and gastric ulcers (RR=0.44; 95% CI:0.26-0.74 and RR=0.37;95% CI;0.27-0.51 respectively for gastric ulcer), and were better tolerated than misoprostol. REVIEWER'S CONCLUSIONS: Misoprostol, PPIs, and double dose H2RAs are effective at preventing chronic NSAID related endoscopic gastric and duodenal ulcers. Lower doses of misoprostol are less effective and are still associated with diarrhea. Only Misoprostol 800ug/day has been directly shown to reduce the risk of ulcer complications. PMID- 11034749 TI - Compression for preventing recurrence of venous ulcers. AB - BACKGROUND: Up to 1% of adults will suffer from leg ulceration at some time. The majority of leg ulcers are venous in origin and are caused by high pressure in the veins due to blockage or weakness of the valves in the veins of the leg. Prevention and treatment of venous ulcers is aimed at reducing the pressure either by removing / repairing the veins, or by applying compression bandages / stockings to reduce the pressure in the veins. The vast majority of venous ulcers are healed using compression bandages. Once healed they often recur and so it is customary to continue applying compression in the form of bandages, tights, stockings or socks in order to prevent recurrence. Compression bandages or hosiery (tights, stockings, socks) are often applied for ulcer prevention. OBJECTIVES: To assess the effects of compression hosiery (socks, stockings, tights) or bandages in preventing the recurrence of venous ulcers. To determine whether there is an optimum pressure/type of compression to prevent recurrence of venous ulcers. SEARCH STRATEGY: Searches of 19 databases including the Cochrane Wounds Group trials register and the Cochrane Controlled Trials Register, handsearching of journals, conference proceedings, and bibliographies up to June 2000. SELECTION CRITERIA: Randomised controlled trials evaluating compression bandages or hosiery for prevention of venous leg ulcers. DATA COLLECTION AND ANALYSIS: Data extraction and assessment of study quality were undertaken by two reviewers independently. MAIN RESULTS: No trials compared recurrence rates with and without compression. One trial (300 patients) compared high (UK Class 3) compression hosiery with moderate (UK Class 2) compression hosiery. A intention to treat analysis found no significant reduction in recurrence at five years follow up associated with high compression hosiery compared with moderate compression hosiery (relative risk of recurrence 0.82, 95% confidence interval 0.61 to 1.12). This analysis would tend to underestimate the effectiveness of the high compression hosiery because a significant proportion of people changed from high compression to medium compression hosiery. Compliance rates were significantly higher with medium compression than with high compression hosiery. One trial (166 patients) found no difference in recurrence between two types of medium (UK Class 2) compression hosiery (relative risk of recurrence with Medi was 0.74, 95% confidence interval 0.45 to 1.2). Both trials reported that not wearing compression hosiery was strongly associated with ulcer recurrence and this is circumstantial evidence that compression reduces ulcer recurrence. No trials were found which evaluated compression bandages for preventing ulcer recurrence. REVIEWER'S CONCLUSIONS: No trials compared compression with vs no compression for prevention of ulcer recurrence. Not wearing compression was associated with recurrence in both studies identified in this review. This is circumstantial evidence of the benefit of compression in reducing recurrence. Recurrence rates may be lower in high compression hosiery than in medium compression hosiery and therefore patients should be offered the strongest compression with which they can comply. Further trials are needed to determine the effectiveness of hosiery prescribed in other settings, i.e. in the UK community, in countries other than the UK. PMID- 11034750 TI - Nedocromil sodium vs. sodium cromoglycate for preventing exercise-induced bronchoconstriction in asthmatics. AB - BACKGROUND: Nedocromil sodium and sodium cromoglycate inhaled shortly before exercise appear to reduce the severity of exercise-induced bronchoconstriction. There is some debate over which drug may be more effective. OBJECTIVES: The objective of this review was to compare the effects on post-exercise lung function between prophylactic doses of nedocromil sodium (NSG) and sodium cromoglycate (SCG) in persons diagnosed with exercise-induced bronchoconstriction. SEARCH STRATEGY: Randomized controlled trials were identified from the Cochrane Airways Review Group Asthma Register which compiles searches of CINAHL, EMBASE, MEDLINE and CENTRAL, plus hand searches for trials in 20 journals. Bibliographies of relevant studies and review articles were searched and primary authors, content experts and manufacturers of drugs were contacted for additional relevant studies. No language restrictions were applied. SELECTION CRITERIA: Randomized controlled trials comparing NCS to SCG in prophylactic treatment of exercise-induced bronchoconstriction were eligible. Studies were included if: the participants, aged 6 or over, had a confirmed diagnosis of asthma with exercise-induced bronchoconstriction, were subjected to an exercise challenge sufficient to trigger bronchoconstriction, and the measures of lung function were reported as either changes in forced expiratory volume in one second or peak expiratory flow rate. DATA COLLECTION AND ANALYSIS: Data extraction and methodological quality assessments were conducted independently by two reviewers using standard forms and validated assessment criteria. In some cases results were extrapolated from graphs. Results from similar studies were pooled and reported as the weighted mean difference (WMD) or odds ratio (OR) with 95% confidence intervals (CI) using the random effects model. MAIN RESULTS: Of the 92 citations retrieved from the original search, a total of 8 studies were included in this review (117 participants). No significant difference was noted between NCS and SCG with respect to the maximum percent decrease in FEV1 (WMD = 0.88; 95% CI: -4.50, 2.74), complete protection (i.e. maximum % fall FEV1 still =>10%); OR = 0.95; 95% CI: 0.50 to 1.8, clinical protection (i.e. < 50% improvement over placebo); OR = 0.71; 95% CI: 0.36 to 1.39; unpleasant taste (OR = 6.85; 95% CI: 0.77, 60.73), or sore throat (OR = 3.46; 95% CI: 0.32, 37.48). For these pooled comparisons, no statistically significant heterogeneity was identified. Subgroup analyses based on age, dosage of medications and timing of exercise post-inhalation were consistent with the overall pooled analyses. REVIEWER'S CONCLUSIONS: No significant differences were evident between the effect of NCS and SCG during the immediate post-exercise period in adults and children with EIB with regards to pulmonary function - specifically maximum percent decrease in FEV1, complete protection, clinical protection, or side effects. PMID- 11034751 TI - Influenza vaccine for patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Influenza vaccinations are currently recommended in the care of people with COPD, but these recommendations are based largely on evidence from observational studies with very few randomised controlled trials (RCTs) reported. Influenza infection causes excess morbidity and mortality in COPD patients but there is also the potential for influenza vaccination to cause adverse effects or not to be cost effective. OBJECTIVES: To evaluate the evidence from RCTs for a treatment effect of influenza vaccination in COPD subjects. Outcomes of interest were exacerbation rates, hospitalisations, mortality, lung function and adverse effects. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register and reference lists of articles. References were also provided by a number of drug companies we contacted. SELECTION CRITERIA: RCTs that compared live or inactivated virus vaccines with placebo, either alone or with another vaccine in persons with COPD. Studies of people with asthma were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers extracted data. All entries were double checked. Study authors and drug companies were contacted for missing information. MAIN RESULTS: Nine trials were included but only four of these were specifically performed in COPD patients. The others were conducted on elderly and high-risk individuals, some of whom had chronic lung disease. In one study of inactivated vaccine in COPD patients there was a significant reduction in the total number of exacerbations per vaccinated subject compared with those who received placebo (weighted mean difference (WMD) -0.45, 95% confidence interval -0.75 to -0.15, p = 0.004). This difference was mainly due to the reduction in exacerbations occurring after 3 weeks (WMD -0.44, (95% CI -0.68 to -0.20, p<0.001). The number of patients experiencing late exacerbations was also significantly less (OR= 0.13, 95%CI 0.04 to 0.45, p=0.002). There was no evidence of an effect of intranasal live attenuated virus when this was added to inactivated intramuscular vaccination. In studies in elderly patients (only a minority of whom had COPD), there was a significant increase in the occurrence of local adverse reactions in vaccinees, but the effects were generally mild and transient. REVIEWER'S CONCLUSIONS: It appears, from the limited number of studies performed, that inactivated vaccine may reduce exacerbations in COPD patients. The size of effect was similar to that seen in large observational studies, and was due to a reduction in exacerbations occurring three or more weeks after vaccination. In elderly, high risk patients there was an increase in adverse effects with vaccination, but these are seen early and are usually mild and transient. PMID- 11034752 TI - Inhaled beclomethasone versus placebo for chronic asthma. AB - BACKGROUND: Inhaled beclomethasone diproprionate (BDP) has been, together with inhaled budesonide, the mainstay of anti-inflammatory therapy for asthma for many years. A range of new prophylactic therapies for asthma is becoming available and BDP is now frequently used as the reference treatment against which these newer agents are being compared. OBJECTIVES: The objectives of this review were to: a) Compare the efficacy of BDP with placebo in the treatment of chronic asthma. b) Explore the possibility that a dose response relationship exists for BDP in the treatment of chronic asthma. c) To provide the best estimate of the efficacy of BDP as a benchmark for evaluation of newer asthma therapies. SEARCH STRATEGY: We searched the Cochrane Airways Group Trial Register (1999) and reference lists of articles. We contacted trialists and Glaxo Wellcome for additional studies and searched abstracts of major respiratory society meetings (1997-1999). SELECTION CRITERIA: Randomised trials in children and adults comparing BDP to placebo in the treatment of chronic asthma. Two reviewers independently assessed articles for inclusion and methodological quality. DATA COLLECTION AND ANALYSIS: One reviewer extracted data; authors were contacted to clarify missing information. Quantitative analyses where undertaken using Review Manager (Revman) 4.0.3 with Metaview 3.1. MAIN RESULTS: 52 studies were selected for inclusion (3459 subjects). The studies were generally of high methodological quality. In non-oral steroid treated patients, BDP produced significant improvements in a number of efficacy measures compared to placebo including FEV1 weighted mean difference (WMD) 340ml (95% CI 190-500ml); FEV1 (% predicted) WMD 6% (95% CI 0.4 to 11.5%) and morning PEFR WMD 50 L/min (95% CI 8 to 92 L/min). BDP also led to reductions in rescue beta2 agonist use compared to placebo WMD 1.75 puffs/d (95% CI 1.4 to 2.4 puffs/d) and reduced the likelihood of trial withdrawal due to asthma exacerbation relative risk (RR) 0.26 (95% CI 0.15 to 0.43). In oral steroid treated patients BDP led to significantly greater reductions in oral prednisolone use WMD 5 mg/d (95% CI 4 to 6 mg/d) and a higher likelihood of discontinuing oral prednisolone RR 0.54 (95% CI 0.43 to 0.67). There was little evidence for a clincially worthwhile dose response effect, but few studies recruited patients with more severe asthma. REVIEWER'S CONCLUSIONS: This review has quantified the efficacy of BDP in the treatment of chronic asthma and strongly supports its use. Current asthma guidelines recommend titration of dose to individual patient response, but the published data provide little support for dose titration above 400 mcg/d in patients with mild to moderate asthma. There are insufficient data to draw any conclusions concerning dose-response in patients with severe disease. PMID- 11034753 TI - Addition of intravenous aminophylline to beta2-agonists in adults with acute asthma. AB - BACKGROUND: Aminophylline has been used extensively in acute asthma, but its role is unclear especially with respect to any additional benefit when added to beta2 agonists. OBJECTIVES: To determine the magnitude of effect of the addition of intravenous aminophylline to beta2-agonists in adult patients with acute asthma treated in the emergency setting. SEARCH STRATEGY: Studies were identified from the following sources: The Cochrane Airways Group register (derived from MEDLINE, EMBASE, CINAHL standardised searches), hand searched respiratory journals and meeting abstracts. Potentially relevant articles were obtained, and their bibliographic lists were hand searched for additional articles. The search included searches of the database up to 1999. SELECTION CRITERIA: Randomised controlled trials comparing intravenous aminophylline versus placebo in adults with acute asthma and treated with beta-adrenergic agonists. Patients could be treated with or without corticosteroids or other bronchodilators. DATA COLLECTION AND ANALYSIS: A total of 210 abstracts were identified. Two independent reviewers selected a total of 27 eligible studies for possible inclusion, in which quality assessment was performed and a third reviewer was used to adjudicate disagreements. Peak expiratory flow (PEFR) and forced expiratory volume in the first second (FEV1) data were extracted and entered in Review Manager from these studies. Information not obtained from the authors was estimated from graphs. All data were entered and double checked by two reviewers. Results are reported as weighted mean differences (WMD) or odds ratio (OR), both with 95% confidential intervals (CI). MAIN RESULTS: Fifteen trials were included. Overall, the quality of the studies was only moderate; concealment of allocation was assessed as clearly adequate in only seven (45%) of the trials. The doses of aminophylline and other medications and the severity of asthma varied between studies. There was no statistically significant effect of aminophylline on airflow outcomes at any time period. The aminophylline treated group had higher values of PEFR at 12 (PEFR 8 L/min or 2.3%) and 24 hours (PEFR 22 L/min or 6.4%), but these were not significant (p>0.05). Two subgroup analyses were performed by grouping studies according to mean baseline airflow limitation (n = 11 studies) and the use of any steroids (n = 9 studies). There was no relationship between baseline airflow limitation nor the use of steroids on the effect of aminophylline. Aminophylline treated patients reported more palpitations/arrhythmias (OR: 2.9; 95% CI: 1.5 to 5.7) and vomiting (OR: 4.2; 95% CI 2.4 to 7.4), but no difference was found in tremor or hospital admissions. REVIEWER'S CONCLUSIONS: In acute asthma, the use of intravenous aminophylline did not result in any additional bronchodilation compared to standard care with beta-agonists. The frequency of adverse effects was higher with aminophylline. No subgroups in which aminophylline might be more effective could be identified. These results should be added to consensus statements and guidelines. PMID- 11034754 TI - Single dose dihydrocodeine for acute postoperative pain. AB - BACKGROUND: Dihydrocodeine is a synthetic opioid analgesic developed in the early 1900s. Its structure and pharmacokinetics are similar to that of codeine and it is used for the treatment of postoperative pain or as an antitussive. It is becoming increasingly important to assess the relative efficacy and harm caused by different treatments. Relative efficacy can be determined when an analgesic is compared with control under similar clinical circumstances. OBJECTIVES: To quantitatively assess the analgesic efficacy and adverse effects of single-dose dihydrocodeine compared with placebo in randomised trials in moderate to severe postoperative pain. SEARCH STRATEGY: Published reports were identified from a variety of electronic databases including Medline, Biological Abstracts, Embase, the Cochrane Library and the Oxford Pain Relief Database. Additional studies were identified from the reference lists of retrieved reports. SELECTION CRITERIA: The following inclusion criteria were used: full journal publication, clinical trial, random allocation of patients to treatment groups, double blind design, adult patients, pain of moderate to severe intensity at baseline, postoperative administration of study drugs, treatment arms which included dihydrocodeine and placebo and either oral or injected (intramuscular or intravenous) administration of study drugs. DATA COLLECTION AND ANALYSIS: Data collection and analysis: Summed pain intensity and pain relief data over 4-6 hours were extracted and converted into dichotomous information to yield the number of patients obtaining at least 50% pain relief. This was used to calculate relative benefit and number needed-to-treat for one patient to obtain at least 50% pain relief. Single-dose adverse effect data were collected and used to calculate relative risk and number needed-to-harm. MAIN RESULTS: Fifty-two reports were identified as possible randomised trials which assessed dihydrocodeine in postoperative pain. Four reports met the inclusion criteria; all assessed oral dihydrocodeine. Three reports (194 patients) compared dihydrocodeine with placebo and one (120 patients) compared dihydrocodeine (30 mg or 60 mg) with ibuprofen 400 mg. For a single dose of dihydrocodeine 30 mg in moderate to severe postoperative pain the NNT for at least 50% pain relief was 8.1 (95% confidence interval 4.1 to 540) when compared with placebo over a period of 4-6 hours. Pooled data showed significantly more patients to have reported adverse effects with dihydrocodeine 30 mg than with placebo. When compared to ibuprofen 400 mg both dihydrocodeine 30 mg and 60 mg were significantly inferior. REVIEWER'S CONCLUSIONS: A single 30 mg dose of dihydrocodeine is not sufficient to provide adequate pain relief in postoperative pain. Statistical superiority of ibuprofen 400 mg over dihydrocodeine (30 mg or 60 mg) was shown. PMID- 11034755 TI - Single dose piroxicam for acute postoperative pain. AB - BACKGROUND: Piroxicam is a non-steroidal anti-inflammatory drug (NSAID) with analgesic properties, and is used mainly for treating rheumatic disorders. Some drugs have been directly compared against each other within a trial setting to determine their relative efficacies, whereas other have not. It is possible, however, to compare analgesics indirectly by examining the effectiveness of each drug against placebo when used in similar clinical situations. OBJECTIVES: To determine the analgesic efficacy and adverse effects of single-dose piroxicam compared with placebo in moderate to severe postoperative pain. To compare the effects of piroxicam with other analgesics. SEARCH STRATEGY: Published reports were identified from systematic searching of Medline, Biological Abstracts, Embase, The Cochrane Library and the Oxford Pain Relief Database. Additional studies were identified from the reference lists of retrieved reports. SELECTION CRITERIA: The following inclusion criteria were used: full journal publication, randomised placebo controlled trial, double-blind design, adult patients, postoperative pain of moderate to severe intensity at the baseline assessment, postoperative administration of oral or intramuscular piroxicam. DATA COLLECTION AND ANALYSIS: Summed pain intensity and pain relief data were extracted and converted into dichotomous information to yield the number of patients obtaining at least 50% pain relief. This was used to calculate estimates of relative benefit and number-needed-to-treat for one patient to obtain at least 50% pain relief. Information was collected on adverse effects and estimates of relative risk and number-needed-to-harm were calculated. MAIN RESULTS: Three trials (141 patients) compared oral piroxicam 20 mg with placebo and one (15 patients) compared oral piroxicam 40 mg with placebo. For single doses of piroxicam 20 mg and 40 mg the respective numbers-needed-to-treat for at least 50% pain relief were 2.7 (2.1 to 3.8) [95% confidence interval] and 1.9 (1.2 to 4.3) [95% confidence interval] compared with placebo over 4-6 hours in moderate to severe postoperative pain. The reported incidence of adverse effects was no higher with piroxicam (20 mg or 40 mg) than with placebo. REVIEWER'S CONCLUSIONS: Piroxicam appears to be of similar efficacy to other non-steroidal anti-inflammatory drugs (NSAIDs) and intramuscular morphine 10 mg when used as a single oral dose in the treatment of moderate to severe postoperative pain. PMID- 11034756 TI - Single dose oxycodone and oxycodone plus paracetamol (acetominophen) for acute postoperative pain. AB - BACKGROUND: Oxycodone is a strong opioid agonist which is useful for the management of severe pain. It is becoming increasingly important to assess the relative efficacy and harm caused by different treatments. This can be determined when an analgesic is compared with control under similar clinical circumstances. OBJECTIVES: To quantitatively assess the analgesic efficacy and adverse effects of single-dose oxycodone and oxycodone plus paracetamol in randomised trials in acute postoperative pain. SEARCH STRATEGY: Published reports were identified from Medline, Biological Abstracts, Embase, the Cochrane Library and the Oxford Pain Relief Database. Additional studies were identified from the reference lists of retrieved reports. SELECTION CRITERIA: The inclusion criteria were: full journal publication, clinical trial, random allocation of adult patients to treatment groups, double blind design, moderate to severe baseline pain, postoperative administration of study drugs, treatment arms which included oxycodone or oxycodone plus paracetamol and placebo (or active control for which comparable efficacy data exist), and oral, intramuscular or intravenous administration of study drugs. DATA COLLECTION AND ANALYSIS: Summed pain intensity and pain relief data over 4-6 hours were extracted and converted into dichotomous information yielding the number of patients obtaining at least 50% pain relief. Estimates of relative benefit and number-needed-to-treat were calculated. Single-dose adverse effect data were collected. MAIN RESULTS: Seventy-seven reports were identified. Seven reports met the inclusion criteria; all assessed oral oxycodone. For efficacy, a significant benefit of active drug over placebo was shown for all doses of oxycodone and oxycodone plus paracetamol, except oxycodone 5 mg. For adverse effects, the number of patients reporting adverse effects was extracted for each dose of active drug versus placebo. When these data were pooled for the individual doses significantly more adverse effects with active drug than with placebo were shown for all doses, except oxycodone 5 mg and its combination with paracetamol 325 mg. This was also shown for drowsiness/somnolence. Significantly more nausea, vomiting and dizziness/lightheadedness were reported with oxycodone 10 mg plus paracetamol (650 mg and 1000 mg) than with placebo. REVIEWER'S CONCLUSIONS: Single-dose oral oxycodone, with or without paracetamol, appears to be of comparable efficacy to intramuscular morphine and non-steroidal anti inflammatory drugs. Central nervous system adverse effects were common. PMID- 11034757 TI - Surgery for the resolution of symptoms in malignant bowel obstruction in advanced gynaecological and gastrointestinal cancer. AB - BACKGROUND: Intestinal obstruction commonly occurs in progressive advanced gynaecological and gastrointestinal cancers. Management of these patients is difficult due to the patients deteriorating mobility and function (performance status), the lack of further chemotherapeutic options and the high mortality and morbidity associated with palliative surgery. There are marked variations in clinical practice concerning surgery in these patients between different countries, gynaecological oncology units, and general hospitals as well as referral patterns from oncologists under whom these patients are often admitted under. There is therefore a need for all the present information to be collated, analysed (with appropriate palliative care outcomes) to establish if surgery is of benefit and what further research is needed. OBJECTIVES: The objective was to locate, appraise and summarise evidence from scientific studies on intestinal obstruction due to advanced gynaecological and gastrointestinal cancer, in order to assess the efficacy of surgery. SEARCH STRATEGY: A comprehensive list of studies was provided by an extensive search of electronic databases, relevant journals, bibliographic databases, conference proceedings, reference lists, the grey literature, personal contact and the world wide web. SELECTION CRITERIA: As the review concentrates on the 'best evidence' available of the role of surgery in malignant bowel obstruction in advanced gynaecological and gastrointestinal cancer the inclusion criteria were kept broad (included both prospective and retrospective studies) so as to include all studies relevant to the question. DATA COLLECTION AND ANALYSIS: Data extraction forms were used to collect data from the studies included in the review. Two researchers extracted the data independently to reduce error. Due to the methodological quality of the studies, only a qualitative assessment was possible. MAIN RESULTS: The role of surgery in malignant bowel obstruction remains controversial, and no firm conclusions from the many retrospective case series can be made. Control of symptoms varies from 42% to over 80%, though it is often unclear how symptoms were measured and whether the tools used to collect symptom scores are validated. There is a large range in the rates of re-obstruction, from 10-50%, though time to re-obstruction was often not included. There is a wide range of postoperative morbidity and mortality, although again the definition of both these surgical outcomes varied between many of the papers. REVIEWER'S CONCLUSIONS: The role of surgery in malignant bowel obstruction needs careful evaluation, using validated outcome measures of symptom control and quality of life scores. Further information would include re-obstruction rates together with the morbidity associated with the various surgical procedures. Currently, bowel obstruction is managed empirically, and there are marked variations in clinical practice by different units. There needs to be a greater standardisation of management so that comparisons between different series can be made. PMID- 11034758 TI - Effects of nonsteroidal anti-inflammatory drugs on post-operative renal function in adults. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) can play a major role in the management of acute pain in the peri-operative period. However, there are conflicting views on whether NSAIDs are associated with adverse renal effects. OBJECTIVES: The primary objective of this review was to determine the effects of NSAIDs on post-operative renal function in adults. SEARCH STRATEGY: Electronic searches for relevant randomised and quasi-randomised controlled trials in Cochrane Controlled Trials Register, MEDLINE and EMBASE were performed. Attempts were also made to identify trials from citation lists of relevant trials, review articles and clinical practice guidelines. Hand-searching of conference abstracts published in major anaesthetic journals was also performed. SELECTION CRITERIA: The inclusion criteria were randomised or quasi-randomised comparisons of individual NSAIDs with either each other or placebo for treatment of post operative pain, with relevant post-operative renal outcome measures, in adult surgical patients. DATA COLLECTION AND ANALYSIS: Of the 14 trials that fulfilled the selection criteria for this review, eight trials were relevant with sufficient data for meta-analysis. The data was extracted independently by two reviewers. The primary outcome measure was creatinine clearance within the first two days after surgery. Secondary outcome measures included serum creatinine, urine volume, urinary sodium level, urinary potassium level, fractional excretion of sodium, fractional excretion of potassium, need for dialysis and need for diuretic or dopamine treatment for renal insufficiency. Weighted mean differences for continuous outcomes and relative risk for dichotomous outcomes were estimated. MAIN RESULTS: As a group, NSAIDs reduced creatinine clearance by 18ml/min (95%CI: 6 to 31) and potassium output by 38mmol/day (95%CI: 19 to 56) on the first day after surgery compared to placebo. Serum creatinine clearance increased on the second day after surgery by 15umol/L (95%CI: 2 to 28) compared to placebo. No significant reduction in urine volume during the early post operative period was found. There was no significant difference in serum creatinine in the early post-operative period between patients receiving ketorolac and diclofenac in one trial. No cases of post-operative renal failure requiring dialysis were described. REVIEWER'S CONCLUSIONS: NSAIDs caused a clinically unimportant transient reduction in renal function in the early post operative period. NSAIDs should not be withheld from adults with normal pre operative renal function because of concerns about post-operative renal impairment. PMID- 11034759 TI - Kangaroo mother care to reduce morbidity and mortality in low birthweight infants. AB - BACKGROUND: Kangaroo mother care (KMC), defined as skin-to-skin contact between a mother and her newborn, frequent and exclusive or nearly exclusive breastfeeding, and early discharge from hospital, has been proposed as an alternative to conventional neonatal care for low birthweight (LBW) infants. OBJECTIVES: To determine whether there is evidence to support the use of KMC in LBW infants as an alternative to conventional care after the initial period of stabilization with conventional care. SEARCH STRATEGY: We used the standard search strategy of the Neonatal Review Group of the Cochrane Collaboration. MEDLINE, EMBASE, LILACS, POPLINE and CINAHL databases, and the Cochrane Controlled Trials Register (Cochrane Library) up to Issue 2, 2000, were searched using the key words terms "kangaroo mother care" or "kangaroo mother method" or "skin-to-skin contact" and "infants" or "low birthweight infants". SELECTION CRITERIA: Randomised trials comparing KMC and conventional neonatal care in LBW infants. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by two reviewers. Statistical analysis was conducted using the standard Cochrane Collaboration methods. MAIN RESULTS: Three studies, involving 1362 infants, were included. All the trials were conducted in developing countries. The studies were of moderate to poor methodological quality. The most common shortcomings were in the areas of blinding procedures for those who collected the outcomes measures, handling of drop outs, and completeness of follow-up. The great majority of results consist of results of a single trial. KMC was associated with the following reduced risks: nosocomial infection at 41 weeks' corrected gestational age (relative risk 0.49, 95% confidence interval 0.25 to 0.93), severe illness (relative risk 0.30, 95% confidence interval 0.14 to 0.67), lower respiratory tract disease at 6 months follow-up (relative risk 0.37, 95% confidence interval 0.15 to 0.89), not exclusively breastfeeding at discharge (relative risk 0.41, 95% confidence interval 0.25 to 0.68), and maternal dissatisfaction with method of care (relative risk 0.41, 95% confidence interval 0.22 to 0.75). KMC infants had gained more weight per day by discharge (weighted mean difference 3.6 g/day, 95% confidence interval 0.8 to 6.4). Scores on mother's sense of competence according to infant stay in hospital and admission to NICU were better in KMC than in control group (weighted mean differences 0.31 [95% confidence interval 0.13 to 0.50] and 0.28 [95% confidence interval 0.11 to 0.46], respectively). Scores on mother's perception of social support according to infant stay in NICU were worse in KMC group than in control group (weighted mean difference -0.18 (95% confidence interval -0.35 to -0.01). There was no evidence of a difference in infant mortality. However, serious concerns about the methodological quality of the included trials weaken credibility in these findings. REVIEWER'S CONCLUSIONS: Although KMC appears to reduce severe infant morbidity without any serious deleterious effect reported, there is still insufficient evidence to recommend its routine use in LBW infants. Well designed randomized controlled trials of this intervention are needed. PMID- 11034760 TI - Neuromuscular paralysis for newborn infants receiving mechanical ventilation. AB - BACKGROUND: Ventilated newborn infants breathing in asynchrony with the ventilator are at risk for complications during mechanical ventilation, such as pneumothorax or intraventricular hemorrhage, and are exposed to more severe barotrauma, which consequently could impair their clinical outcome. Neuromuscular paralysis, which eliminates spontaneous breathing efforts of the infant, has potential advantages in this respect. However, a number of complications have been reported with muscle relaxation in infants, so that concerns exist regarding the safety of prolonged neuromuscular paralysis in newborn infants. OBJECTIVES: To determine whether routine neuromuscular paralysis of newborn infants receiving mechanical ventilation compared with no routine paralysis results in clinically important benefits or harms. SEARCH STRATEGY: MEDLINE (from 1966 to May 2000) and EMBASE (from 1988 to May 2000) were searched, as well as The Cochrane Controlled Trials Register (issue 2, 2000). References of review articles were hand searched. Language restriction was not imposed. SELECTION CRITERIA: All trials using random or quasi-random patient allocation, in which the routine use of neuromuscular blocking agents during mechanical ventilation was compared to no paralysis or selective paralysis in newborn infants. Methodological quality was assessed blindly and independently by the two authors. DATA COLLECTION AND ANALYSIS: Data were abstracted using standard methods of the Cochrane Collaboration and its Neonatal Review Group, with independent evaluation of trial quality, and abstraction and synthesis of data by both authors. Treatment effect was analysed using relative risk, risk difference and weighted mean difference. MAIN RESULTS: Ten possibly eligible trials were identified, of which five were included in the review. All the included trials studied preterm infants ventilated for respiratory distress syndrome, and used pancuronium as the neuromuscular blocking agent. In the analysis of the results of all trials, no difference was found in mortality, air leak or chronic lung disease, but there was a significant reduction in intraventricular hemorrhage and a trend towards less severe intraventricular hemorrhages. In the subgroup analysis of trials studying a selected population of ventilated infants with evidence of asynchronous respiratory efforts, a significant reduction in intraventricular hemorrhage (any grade and severe IVH) was found, and a trend towards less air leak. In the subgroup analysis of trials studying an unselected population of ventilated infants, no differences were found for any of the outcomes. REVIEWER'S CONCLUSIONS: For ventilated preterm infants with evidence of asynchronous respiratory efforts, neuromuscular paralysis with pancuronium seems to have a favourable effect on intraventricular hemorrhage and possibly on air leak. Uncertainty remains, however, regarding the long term pulmonary and neurologic effects, and regarding the safety of prolonged use of pancuronium in ventilated newborn infants. There is no evidence from randomized trials on the effects of neuromuscular blocking agents other than pancuronium. Therefore, the routine use of pancuronium or any other neuromuscular blocking agent in ventilated newborn infants cannot be recommended based on current evidence. PMID- 11034761 TI - Prophylactic vitamin K for vitamin K deficiency bleeding in neonates. AB - BACKGROUND: Vitamin K deficiency can cause bleeding in an infant in the first weeks of life. This is known as Hemorrhagic Disease of the Newborn (HDN). HDN is divided into three categories: early, classic and late HDN. Early HDN occurs within 24 hours post partum and falls outside the scope of this review. Classic HDN occurs on days one to seven; common bleeding sites are gastrointestinal, cutaneous, nasal and from a circumcision. Late HDN occurs from week 2-12; the most common bleeding sites are intracranial, cutaneous, and gastrointestinal. Vitamin K is commonly given prophylactically after birth for the prevention of HDN, but the preferred route is uncertain. OBJECTIVES: To review the evidence from randomized trials in order to determine the effectiveness of vitamin K prophylaxis in the prevention of classic and late HDN. Main questions are: Is one dose of vitamin K, given after birth, able to significantly reduce the incidence of classic and late HDN? Is there a significant difference between the oral route and the intramuscular route in preventing classic and late HDN? Are multiple oral doses of vitamin K, given after birth, able to significantly reduce the incidence of classic and late HDN? SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. SELECTION CRITERIA: All trials using random or quasi-random patient allocation, in which methods of vitamin K prophylaxis in infants were compared to each other, placebo or no treatment, were included. DATA COLLECTION AND ANALYSIS: Data were extracted independently by each author and were analysed with the standard methods of the Cochrane Collaboration and its Neonatal Review Group, using relative risk, risk difference and weighted mean difference. MAIN RESULTS: Two eligible randomized trials, each comparing a single dose of intramuscular vitamin K with placebo or nothing, assessed effect on clinical bleeding. One dose of vitamin K reduced clinical bleeding at 1-7 days, including bleeding after circumcision, and improved biochemical indices of coagulation status. Eleven additional eligible randomized trials compared either a single oral dose of vitamin K with placebo or nothing, a single oral with a single intramuscular dose of vitamin K, or three oral doses with a single intramuscular dose. None of these trials assessed clinical bleeding. Oral vitamin K improved biochemical indices of coagulation status at 1-7 days. There was no evidence of a difference between the oral and intramuscular route in effects on biochemical indices of coagulation status. A single oral compared with a single intramuscular dose resulted in lower plasma vitamin K levels at two weeks and one month, whereas a 3-dose oral schedule resulted in higher plasma vitamin K levels at two weeks and at two months than did a single intramuscular dose. REVIEWER'S CONCLUSIONS: A single dose (1.0 mg) of intramuscular vitamin K after birth is effective in the prevention of classic HDN. Either intramuscular or oral (1.0 mg) vitamin K prophylaxis improves biochemical indices of coagulation status at 1-7 days. Neither intramuscular nor oral vitamin K has been tested in randomized trials with respect to effect on late HDN. Oral vitamin K, either single or multiple dose, has not been tested in randomized trials for its effect on either classic or late HDN. PMID- 11034762 TI - Surgery versus thrombolysis for acute limb ischaemia: initial management. AB - BACKGROUND: Peripheral arterial thrombolysis has become established as a useful adjunct in the management of peripheral arterial ischaemia. Much has been learnt about indications, risks and benefits using this technique, although data from randomised controlled studies is not extensive. The optimal initial management of the acutely ischaemic leg needs to be determined. OBJECTIVES: To determine if surgery or thrombolysis is the preferred option in the initial treatment of acute limb ischaemia. SEARCH STRATEGY: The search strategy was that adopted by the Cochrane Review Group on Peripheral Vascular Diseases. Additionally, reference lists of papers resulting from this search were reviewed. SELECTION CRITERIA: All randomised studies comparing thrombolysis and surgery in the management of acute limb ischaemia. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted independently by all three reviewers. MAIN RESULTS: Patients with acute lesions of less than seven days duration had a significantly increased survival at one year for patients having thrombolysis, compared to those undergoing initial surgery [84% v 58%, p=0.01; Odds ratio (95% CI) 0.28 (0.13,0.63)] largely associated with a reduced level of in-hospital cardio pulmonary complications (Ouriel 1994). Lesions less than 14 days duration fared better with initial lysis with a reduced amputation and reduced death rate at six months [15.3% v 37.5%; p=0.001; Odds ratio (95%CI) 0.29 (0.12,0.72)] (STILE 1994), compared to initial surgery. Analysis of the same trial at one year however revealed that native vessel thromboses had a more favourable outcome with initial surgery, largely due to continuing ischaemia in the lytic group [64% v 35%; p<0.0001; Odds ratio (95%CI) 3.26(1.96,5.52)] (Weaver 1996). Bypass graft thromboses less than 14 days old treated with initial thrombolysis were shown to have a reduced amputation rate (15% v 47%; p=0.05). However, overall, one year results revealed that thrombolysis of thrombosed grafts was associated with a higher level of continued ischaemia [73% v 50%; P=0.010; Odds ratio (95%CI) 2.72(1.27,5.80)] (Comerota 1996). REVIEWER'S CONCLUSIONS: A universal initial treatment with either surgery or thrombolysis cannot be advocated on the available evidence. There is no overall difference in limb salvage or death at one year between initial surgery and initial thrombolysis. Thrombolysis may however be associated with a higher risk of ongoing limb ischaemia, and a higher overall risk of haemorrhagic complications including stroke. The higher risk of complications needs to be balanced against the risks of surgery in the individual patient. PMID- 11034763 TI - Inhaled nitric oxide for acute hypoxemic respiratory failure in children and adults. AB - OBJECTIVES: To determine the effect of iNO, compared with inhaled placebo, on outcome in AHRF in children and/or adults. SEARCH STRATEGY: Randomised controlled trials (RCT's) were identified from electronic databases; MEDLINE, EMBASE, the Cochrane Library, and CINAHL, as well as from bibliographies of retrieved articles. Relevant journals and conference proceedings were hand searched and authors published in this field were contacted for knowledge of unpublished ongoing trials. SELECTION CRITERIA: RCT's comparing iNO with maximal conventional therapy and inhaled placebo, for AHRF in either children or adults. DATA COLLECTION AND ANALYSIS: Data were extracted and the analyses performed independently by two reviewers. All 5 authors were contacted for missing data. Qualitative assessment of each trial was made according to methodology described by Schulz (Schulz 1995), and analyses according to statistical methods in Review Manager MetaView 3.1. The fixed effect model was applied. Where possible, sub group analyses were performed to assess the impact of iNO in varied doses. MAIN RESULTS: Five RCT's were evaluated, assessing 535 patients with AHRF. Inhaled nitric oxide made no impact on mortality in trials without cross-over (RR 0.98, 95%CI 0.66, 1.44), or with cross-over of treatment failures to open-label iNO (RR 1.22, 95%CI 0.65, 2.29). Published evidence from one study demonstrated that iNO resulted in a transient improvement in oxygenation in the first 24 hours of treatment: the oxygenation index (OI) showed a mean difference of -3 [95% CI 5.354, -0.646], and PaO2/FiO2 ratio, a mean difference of 35 [95% CI 20.236, 49.764]. Limited data demonstrated no difference in ventilator-free days between treatment and placebo groups, and no specific dose of iNO was significantly advantageous over another. Other clinical indicators of effectiveness, such as duration of hospital and intensive care stay, were inconsistently reported. There were no complications reported to be directly attributable to this treatment. REVIEWER'S CONCLUSIONS: From the data provided to date, iNO had no effect on mortality and only transiently improved oxygenation in AHRF in children and/or adults. There was a lack of data to assess other end points. The long term adverse effects of this drug are not known, as no long term follow-up of trial participants has been reported. If further trials comparing iNO with an inhaled placebo are to proceed, they should be stratified for primary disease and must specifically evaluate clinically relevant outcomes, before any benefit of iNO in AHRF can be excluded. PMID- 11034764 TI - Selective serotonin reuptake inhibitors versus tricyclic and heterocyclic antidepressants: comparison of drug adherence. AB - BACKGROUND: Selective serotonin reuptake inhibitors are thought to have better discontinuation rates (i.e. less people dropping out) than tricyclic and heterocyclic antidepressant drugs. It is important to quantify the drop-out rates of different antidepressant drugs in order to have a better understanding of the relative tolerability of these drugs. OBJECTIVES: To assess the comparative tolerability of selective serotonin reuptake inhibitors and tricyclic/heterocyclic antidepressant drugs. SEARCH STRATEGY: We searched the Cochrane Collaboration Depression, Anxiety and Neurosis Controlled Trials Registers (1997 to 1999), MEDLINE (1966 to 1999), EMBASE (1974 to 1999) We also searched specialist journals, the reference lists of relevant papers and previous systematic reviews, conference abstracts and government documents. Representatives of the pharmaceutical industry were contacted. SELECTION CRITERIA: Parallel group randomised controlled trials comparing selective serotonin reuptake inhibitors with tricyclic or heterocyclic antidepressants in people with depression. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and a third reviewer checked any cases of disagreement. MAIN RESULTS: We included 136 trials. The selective serotonin reuptake inhibitors showed less participants dropping out compared to the tricyclic/heterocyclic group (odds ratio 1.21, 95% confidence interval 1.12 to 1.30). A statistically significant difference was found in total drop-outs between the selective serotonin reuptake inhibitors and the old tricyclics as well as the newer tricyclics. When the selective serotonin reuptake inhibitors were compared to the heterocyclic antidepressants, there was a non significant difference favouring the selective serotonin reuptake inhibitors. The poor tolerability profile of the old tricyclics was explained by differences in drop-outs for side-effects, but not for inefficacy. REVIEWER'S CONCLUSIONS: Whilst selective serotonin reuptake inhibitors do appear to show an advantage over tricyclic drugs in terms of total drop-outs, this advantage is relatively modest. This has implications for pharmaco-economic models, some of which may have overestimated the difference of drop-out rates between selective serotonin reuptake inhibitors and tricyclic antdepressants. These results are based on short-term randomised controlled trials, and may not generalise into clinical practice. PMID- 11034765 TI - Pharmacotherapy for posttraumatic stress disorder. AB - BACKGROUND: Posttraumatic stress disorder (PTSD) is a prevalent and disabling disorder. By definition prior psychological trauma plays a causal role in the disorder, and psychotherapy is a widely accepted intervention. Nevertheless there is growing evidence that PTSD is characterized by specific psychobiological dysfunctions, and this has contributed to a growing interest in the use of medication in its treatment. OBJECTIVES: The authors aimed to undertake a systematic review of randomized controlled trials (RCTs) of the pharmacotherapy of posttraumatic stress disorder (PTSD) following the guidelines and using the software of the Cochrane Collaboration, and to provide an estimate of the effects of medication in this disorder. Secondary objectives were to explore questions about whether particular classes of medication are more effective and/or acceptable than others in the treatment of PTSD, and about which factors (clinical and methodological) predict response to pharmacotherapy. SEARCH STRATEGY: Studies of the pharmacotherapy of PTSD were identified using literature searches of MEDLINE (1966 to 1999, using the textwords posttraumatic, post traumatic, medication, pharmacotherapy) and other electronic databases (PSYCLIT; National PTSD Center Pilots database; Dissertation Abstracts; Cochrane Collaboration Depression, Anxiety & Neurosis Controlled Trials Register). In addition, published and unpublished RCTs were requested from PTSD researchers and pharmaceutical companies. An initial broad strategy was undertaken to find not only RCTs, but also open-label trials and reviews of the pharmacotherapy of PTSD; additional studies were sought in reference lists of retrieved articles and included studies in any language. SELECTION CRITERIA: All RCTs of PTSD (including both placebo controlled and comparative trials), whether published or unpublished, but completed prior to the end of 1999 were considered for the review. DATA COLLECTION AND ANALYSIS: Selected RCTs were independently assessed and collated by 2 raters, and Review Manager (RevMan) software was used to capture data on treatment response and PTSD symptom ratings. Ratings of subtypes of PTSD symptoms (intrusive/re-experiencing, avoidant/numbing, hyperarousal), of comorbid symptoms (depression, anxiety), and of quality of life were included where possible. A range of additional information, which may explain possible clinical and methodological heterogeneity amongst the trials, was also captured. Summary statistics for dichotomous and continous measures were calculated, heterogeneity was assessed, and subgroup/sensitivity analyses undertaken. MAIN RESULTS: 15 short-term (12 weeks or less) RCTs were found, of which 9 had sufficient data for inclusion in the analysis. Methodological limitations were particularly apparent in early work; these included short (5 weeks or less) duration of trials and a reliance on self-report scales (rather than use of standardized clinician-rated scales). Despite these and other potentially important differences between the trials, many trials demonstrated efficacy for medication over placebo. Summary statistics were calculated for the Clinical Global Impressions scale change item (CGI-C) or close equivalent from 10 sets of data including various antidepressants and other agents - the proportion of non responders was lower in the pharmacotherapy group than in the control group (relative risk (95% CI) = 0.72 (0.64, 0.83)). Similarly, summary statistics for the intrusion, avoidance and total scales of the Impact of Events Scale (IES) from 4sets of data again including various agents showed a statistically (and clinically) significant difference between medication and placebo (Weighted Mean Difference (95% CI) = -3.81 (-6.72,-0.91), -3.31(-5.24,-1.37), -7.18 (-11.86, 2.50) respectively). REVIEWER'S CONCLUSIONS: Medication treatments can be effective in PTSD, acting to reduce its core symptoms, and should be considered as part of the treatment of this disorder. The existing evidence base does not provide sufficient data to suggest particular predictors of response to treatment, or to demonstrate that any particular class of medication is more effective or better tolerated than any other. However, the largest trials showing efficacy to date have been with the SSRIs, and in contrast, there have been negative studies of some agents. Given the high prevalence and enormous personal and societal costs of PTSD, there is a need for additional controlled trials in this area. Additional questions for future research include the effects of medication on quality of life in PTSD, appropriate dose and duration of medication, the use of medication in different trauma groups, in pediatric and geriatric subjects, and the value of early (prophylactic), combined (with psychotherapy), and long-term (maintenance) medication treatment. PMID- 11034766 TI - Cranial irradiation for preventing brain metastases of small cell lung cancer in patients in complete remission. AB - BACKGROUND: Prophylactic cranial irradiation halves the rate of brain metastases in patients with small cell lung cancer. Individual randomized trials conducted on patients in complete remission were unable to clarify whether this treatment improves survival. OBJECTIVES: This study aims to test whether prophylactic cranial irradiation prolongs survival of patients with small cell lung cancer in complete remission. SEARCH STRATEGY: Published and unpublished trials were eligible. Electronic databases, reference lists of trial publications, review articles and relevant books were used to identify potentially eligible trials. The search was also guided by discussions with investigators and experts, and the examination of meeting proceedings and of the Physician Data Query clinical trial registry. SELECTION CRITERIA: Randomized trials comparing prophylactic cranial irradiation with no prophylactic cranial irradiation in patients with small cell lung cancer in complete remission. DATA COLLECTION AND ANALYSIS: Meta-analysis based on updated individual data. The main endpoint was survival. MAIN RESULTS: The relative risk of death in the treatment group compared to the control group was 0.84 (95% confidence interval=0.73 to 0.97, P=0.01), corresponding to a 5.4 percent increase in the 3-year survival rate (from 15.3 percent in the control group to 20.7 percent in the treatment group). Prophylactic cranial irradiation also increased disease-free survival (relative risk=0.75, 95% confidence interval=0.65 to 0.86, P<0.001) and decreased the risk of brain metastases (relative risk=0.46, 95% confidence interval=0.38 to 0.57, P<0.001). Increasing doses of irradiation decreased the risk of brain metastases when four groups (8 Gy, 24-25 Gy, 30 Gy, 36-40 Gy) were analyzed [trend test, P=0.02], but the effect on survival did not differ significantly according to the dose. We found a trend (P=0.01) for a decrease in the brain metastasis risk in favour of earlier administration of cranial irradiation after the initiation of induction treatment. REVIEWER'S CONCLUSIONS: Prophylactic cranial irradiation significantly improves survival and disease-free survival for patients with small cell lung cancer in complete remission. Further clinical trials are needed to confirm the potential greater benefit on brain metastasis rate suggested when cranial irradiation is given earlier or at higher doses. PMID- 11034767 TI - Recombinant versus urinary follicle stimulating hormone for ovarian stimulation in assisted reproduction cycles. AB - BACKGROUND: Until recently, the main source of exogenous follicle stimulating hormone (FSH) for therapeutic use in infertility had been the urine of postmenopausal women. New developments have resulted in the production of FSH in vitro by recombinant DNA technology. The extremely high purity and batch-to-batch consistency of recombinant FSH (rFSH) make it an attractive alternative to urinary FSH (uFSH). OBJECTIVES: To conduct a systematic review and meta-analysis of randomized trials comparing the effectiveness of rFSH with uFSH in ovarian stimulation protocols in in vitro fertilization (IVF) or intra-cytoplasmic sperm injection (ICSI) treatment cycles. SEARCH STRATEGY: Search strategies included on line searching of the MEDLINE and EMBASE databases from 1985 to 1999, hand searching of bibliographies of relevant publications and reviews and abstracts of scientific meetings, peer consultation and contacting the pharmaceutical companies that manufacture the gonadotropins under consideration. SELECTION CRITERIA: Randomized trials comparing rFSH with uFSH for ovarian stimulation in IVF or ICSI treatment for infertility. DATA COLLECTION AND ANALYSIS: The main outcome measure was clinical pregnancy per cycle started. Also considered were clinical pregnancy per cycle reaching oocyte retrieval and per cycle reaching embryo transfer (ET), ongoing pregnancy per cycle started, spontaneous abortion, multiple pregnancy, ovarian hyperstimulation syndrome (OHSS), number of follicles and serum estradiol level on the day of human chorionic gonadotropin administration day, total dose of FSH, and number of oocytes retrieved. Common odds ratios (OR) and risk differences for rFSH relative to uFSH were calculated after testing for homogeneity of treatment effect across all trials. The fixed effects model was used, unless significant heterogeneity was present, in which case the random effects model was used. MAIN RESULTS: The overall odds ratio for clinical pregnancy per cycle started was 1.21 (95% confidence limits (CL) 1.04,1.42) for rFSH compared to uFSH. The risk difference was a 3.7% (0.8,6.7) absolute increase in clinical pregnancy rate with rFSH. The OR for ongoing pregnancy per cycle started was 1.29 (1.08,1.54). There was no significant difference between rFSH and uFSH in the rates of spontaneous abortion, multiple pregnancy or OHSS. The total dose of FSH was lower by 406 (185,627) IU with rFSH, but there was no significant difference in the number of follicles or serum estradiol on hCG day or in the number of oocytes retrieved. REVIEWER'S CONCLUSIONS: This review has demonstrated a statistically significant increase in clinical pregnancy rate with rFSH compared to uFSH, when used for ovarian stimulation in assisted reproduction. This benefit was observed only in standard IVF cycles and not in cycles in which ICSI was used. PMID- 11034768 TI - Transcutaneous electrical nerve stimulation for knee osteoarthritis. AB - BACKGROUND: Osteoarthritis (OA) is a disease that affects synovial joints, which mainly causes degeneration and destruction of hyaline cartilage. To date, no curative treatment for OA exists. The primary goals for OA therapy are to relieve pain, maintain or improve functional status, and minimize deformity. Transcutaneous electrical nerve stimulation (TENS) is a noninvasive modality in physiotherapy that is commonly used to control both acute and chronic pain arising from several conditions. A number of trials evaluating the efficacy of TENS in OA have been published. OBJECTIVES: To assess the effectiveness of TENS in the treatment of knee OA. The primary outcomes of interest were those described by the Outcome Measures in Rheumatology Clinical Trials (OMERACT) 3, which included pain relief, functional status, patient global assessment, and change in joint imaging for studies of one year or longer. The secondary objective was to determine the most effective mode of TENS application in pain control. SEARCH STRATEGY: We searched MEDLINE, EMBASE, CINAHL, HEALTHSTAR, PEDro, Current Contents and the Cochrane Controlled Trial Register using the Cochrane Musculoskeletal Group search strategy for trials up to and including December 1999. We also hand-searched reference lists and consulted content experts. SELECTION CRITERIA: Two independent reviewers selected the trials that met predetermined inclusion criteria. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted the data using standardized forms and assessed the quality of randomization, blinding and dropouts. A third reviewer was consulted to resolve any differences. For dichotomous outcomes, relative risks (RR) were calculated. For continuous data, weighted mean differences (WMD) or standardized mean difference (SMD) of the change from baseline were calculated. A fixed effects model was used unless heterogeneity of the populations existed. In this case, a random effects model was used. MAIN RESULTS: Seven trials were eligible to be included in this review. Six used TENS as the active treatment while one study used acupuncture-like TENS (AL-TENS). A number of 148 and 146 patients were involved in the active TENS treatment and placebo, respectively. Three studies were cross-over studies and the others were parallel group, randomized controlled trials (RCTs). Median methodological quality of these studies was two. Pain relief from active TENS and AL-TENS treatment was significantly better than placebo treatment. Knee stiffness also improved significantly in active treatment group compared to placebo. Different modes of TENS setting (High Rate and Strong Burst Mode TENS) demonstrated a significant benefit in pain relief of the knee OA over placebo. Subgroup analyses showed a heterogeneity in the studies with methodological quality of three or more and those with repeated TENS applications. REVIEWER'S CONCLUSIONS: TENS and AL-TENS are shown to be effective in pain control over placebo in this review. Heterogeneity of the included studies was observed, which might be due to the different study designs and outcomes used. More well designed studies with a standardized protocol and adequate number of participants are needed to conclude the effectiveness of TENS in the treatment of OA of the knee. PMID- 11034769 TI - Fluoride for treating postmenopausal osteoporosis. AB - OBJECTIVES: To assess the efficacy of fluoride therapy on bone loss, vertebral and non-vertebral fractures and side effects in postmenopausal women. SEARCH STRATEGY: We searched Medline, Current Contents and the Cochrane Controlled Trial Registry up to December 1998. SELECTION CRITERIA: Two independent reviewers selected RCTs which met predetermined inclusion criteria. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data using predetermined forms and assessed the methodological quality of the trials using a validated scale. For dichotomous outcomes, relative risks (RR) were calculated and for continuous outcomes, weighted mean differences (WMD) of percentage change from baseline were calculated. Where heterogeneity existed (determined by a chi-square test) a random effects model was used. MAIN RESULTS: Eleven studies (1429 subjects) met the inclusion criteria. The increase in lumbar spine bone mineral density (BMD) was found to be higher in the treatment group than in the control group with a WMD 8.1% (95%CI: 7.15,9.09) after two years of treatment and 16.1%(95%CI: 14.65,17.5) after four years. The RR for new vertebral fractures was not significant at two years [0.87 (95%CI: 0.51,1.46)] or at four years [0.9(95%CI: 0.71,1.14)]. The RR for new non-vertebral fractures was not significant at two years 1.2(95%CI: 0.68,2.1) but was increased at four years in the treated group 1.85(95%CI: 1.36,2.5), especially if used at high doses and in a non slow release form. The RR for gastrointestinal side effects was not significant at two years 2.18(95%CI: 0.86,1.21) but was increased at four years in the treated group 2.18(95%CI: 1.69,4.57) especially if fluoride was used at high doses and in a non slow release form. The number of withdrawals and dropouts was not different between treated and control groups at two and four years. REVIEWER'S CONCLUSIONS: Although fluoride has an ability to increase BMD at lumbar spine, it does not result in a reduction of vertebral fractures. In increasing the dose of fluoride, one increases the risk of non-vertebral fracture and gastrointestinal side effects without any effect on the vertebral fracture rate. PMID- 11034770 TI - Thermotherapy for treating rheumatoid arthritis. AB - BACKGROUND: Heat and cold therapy are often used as adjuncts in the treatment of rheumatoid arthritis by rehabilitation specialists. OBJECTIVES: To evaluate the effects of heat and cold on objective and subjective measures of disease activity in patients with rheumatoid arthritis. SEARCH STRATEGY: We searched Medline, Embase, PEDro, Current Contents, Sports Discus and CINAHL up to June 2000. The Cochrane Field of Rehabilitation and related therapies and the Cochrane Musculoskeletal Review Group were also contacted for a search of their specialized registers. Handsearching was conducted on all retrieved articles for additional articles. SELECTION CRITERIA: Randomized or controlled clinical trials of ice or heat compared to placebo or active interventions in patients with rheumatoid arthritis and case-control and cohort studies were eligible. No language restrictions were applied. Abstracts were accepted. DATA COLLECTION AND ANALYSIS: Two independent reviewers identified potential articles from the literature search. These reviewers extracted data using pre-defined extraction forms. Consensus was reached on all data extraction. Quality was assessed by two reviewers using a 5 point scale that measured the quality of randomization, double-blinding and description of withdrawals. MAIN RESULTS: Three studies (79 subjects) met the inclusion criteria. There was no effect on objective measures of disease activity (including inflammation, pain and x-ray measured joint destruction) of either ice versus control or heat versus control. Patients reported that they preferred heat therapy to no therapy (94% like heat therapy better than no therapy). There was no difference in patient preference for heat or ice. No harmful effects of ice or heat were reported. REVIEWER'S CONCLUSIONS: Since patients enjoy thermotherapy, and there are no harmful effects, thermotherapy should be recommended as a therapy which can be applied at home as needed to relieve pain. There is no need for further research on the effects of heat or cold for RA. PMID- 11034771 TI - Psychoeducation for schizophrenia. AB - BACKGROUND: Schizophrenia can be a severe and chronic illness characterised by lack of insight and poor compliance with treatment. Psychoeducational approaches have been developed to increase patients' awareness of their illness and its treatment. OBJECTIVES: To assess the effects of psychoeducational interventions compared to standard levels of knowledge provision. SEARCH STRATEGY: Electronic searches of CINAHL (1982-1999), The Cochrane Library CENTRAL (Issue 1, 1999), The Cochrane Schizophrenia Group's Register (January 1999), EMbase (1980-1999), MEDLINE (1966-1999), PsycLit (1974-1999), and Sociofile (1974-1999) were undertaken. These were supplemented with reference searching and personal contact with authors of all included studies. SELECTION CRITERIA: All relevant randomised controlled trials focusing on psychoeducation for schizophrenia or related serious mental illnesses, involving individuals or groups. Quasi-randomised trials were excluded. DATA COLLECTION AND ANALYSIS: Data were extracted independently by at least two reviewers from included papers. Authors of trials were contacted for additional and missing data. Relative risks (RR) and 95% confidence intervals (CI) of homogeneous dichotomous data were calculated. A random effects model was used for heterogeneous dichotomous data. Where possible the numbers needed to treat (NNT) were also calculated. Weighted or standardised means were calculated for continuous data. MAIN RESULTS: Ten studies are included in this review. All studies of group education included family members. Compliance with medication was significantly improved in a single study using brief group intervention (at one year) but other studies produced equivocal or skewed data. Any kind of psychoeducational intervention significantly decreased relapse or readmission rates at nine to 18 months follow-up compared with standard care (RR 0.8 CI 0.7-0.9 NNT 9 CI 6-22). Several of the secondary outcomes (knowledge gains, mental state, global level of functioning, status of high expressed emotion family members) were measured using scales that are difficult to interpret. Generally, however, findings were consistent with the possibility that psychoeducation has a positive effect on a persons' well being. No impact was found on insight, medication related attitudes or on overall satisfaction with services of patients or relatives but these findings rested on very few studies. Health economic outcome was only measured in one study and data were skewed. It was not possible to analyse whether different duration or formats of psychoeducation influenced effectiveness. REVIEWER'S CONCLUSIONS: Evidence from trials suggests that psychoeducational approaches are useful as a part of the treatment programme for people with schizophrenia and related illness. That the interventions are brief and inexpensive should make them attractive to managers and policy makers. More well-designed, conducted and reported randomised studies investigating the efficacy of psychoeducation are needed. PMID- 11034772 TI - Vasoactive drugs for acute stroke. AB - BACKGROUND: It is unclear whether blood pressure should be managed after acute stroke and if so whether it is best to reduce or increase blood pressure. OBJECTIVES: The objective of this review was to assess the effect of lowering or elevating blood pressure in people with acute stroke, and the effect of different vasoactive drugs on blood pressure in acute stroke. SEARCH STRATEGY: We searched the Cochrane Library (1999 Issue 1) using the CDSR and the CCTR databases, MEDLINE (from 1966), EMBASE (from 1980), BIDS ISI (Science Citation Index from 1981), and existing review articles. We contacted researchers in the field and pharmaceutical companies. SELECTION CRITERIA: Randomised trials of interventions that would be expected, on pharmacological grounds, to alter blood pressure in patients within two weeks of the onset of acute ischaemic or haemorrhagic stroke. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied the trial inclusion criteria, assessed trial quality, and extracted the data. MAIN RESULTS: Sixty five trials were identified involving in excess of 11,500 patients; a further 5 trials are ongoing. Data were obtained for 32 trials (5,368 patients). Significant imbalances in baseline blood pressure were present across trials of intravenous calcium channel blockers and prostacyclin. Major imbalances in baseline blood pressure between treatment and control groups have made the interpretation of these results difficult. Intravenous calcium channel blockers (CCBs) and oral CCBs significantly lowered late blood pressure as compared to controls. (systolic/diastolic BP): iv CCBs -8.2/-6.7 mm Hg (95% CI -12.6 to 3.8)/ (95% CI -9.2 to -4.3); oral CCBs -3.2/-2.1 mm Hg (95% CI -5.0 to -1.3)/ (95% CI -3.0 to -1.0). Beta blockers significantly lowered late diastolic blood pressure but not significantly late systolic blood pressure; -5.0/-4.5 mm Hg (95% CI -10.2 to 0.4)/(95% CI -7.8 to -1.15). Angiotensin converting enzyme inhibitors and prostacyclin non-significantly reduced late BP as compared to the controls by -5.4/-3.0 mm Hg (95% CI -16.5 to 5.8)/(95% CI -11.1 to 5.0) and -7.4/-3.9 mmHg (95% CI -15.6 to 0.2)/(95% CI -8.1 to 0.4) respectively. Magnesium, naftidrofuryl and piracetam had no significant effect on blood pressure. Oral CCBs and beta blockers each significantly reduced late heart rate (beats per minute (bpm)): CCBs -2.8 bpm (95%CI -3.9 to -1.7); beta blockers -9.3 bpm (95% CI -12.0 to 6.6). Prostacyclin significantly increased late heart rate by +5.6 bpm (95% CI 0.8 to 10.4). None of the drug classes significantly altered outcome apart from beta blockers and streptokinase which increased early case fatality (odds ratio 1.77, 95%CI, 1.05 to 3.00) and 2.27 (95% CI 1.4 to 3.67). REVIEWER'S CONCLUSIONS: There is not enough evidence reliably to evaluate the effect of altering blood pressure on outcome after acute stroke. CCBs, beta blockers, and probably ACE inhibitors, prostacyclin and nitric oxide, each lowered BP during the acute phase of stroke. In contrast, magnesium, naftidrofuryl and piracetam had little or no effect on BP. PMID- 11034773 TI - Cognitive rehabilitation for attention deficits following stroke. AB - BACKGROUND: Attention problems occur following stroke and are treated using computerised activities or paper and pencil tasks. OBJECTIVES: To determine the effects of cognitive rehabilitation for attention deficits following stroke. SEARCH STRATEGY: We searched the Cochrane Stroke Group Trials Register, Medline, EMBASE, CINHAL and CLIN PSYCH databases and reference lists from relevant articles. Date of most recent searches: December 1998 SELECTION CRITERIA: Controlled trials of attention training in stroke. Studies with mixed aetiology groups were excluded unless they included more than 75% of stroke patients or separate data were available for the stroke patients. DATA COLLECTION AND ANALYSIS: Two reviewers extracted trial data and assessed trial quality. Reviewers contacted investigators for further details of trials. MAIN RESULTS: Two trials were identified with 56 participants. The two trials showed a benefit of training on measures of alertness and sustained attention. Only one trial included a measure of functional independence and this showed no significant effect of training. REVIEWER'S CONCLUSIONS: There is some indication that training improves alertness and sustained attention but no evidence to support or refute the use of cognitive rehabilitation for attention deficits to improve functional independence following stroke. PMID- 11034774 TI - Anxiolytics for smoking cessation. AB - BACKGROUND: There are two reasons to believe anxiolytics might help in smoking cessation. Anxiety may be a symptom of nicotine withdrawal. Second, smoking appears to be due, in part, to deficits in dopamine, serotonin and norepinephrine, all of which are increased by anxiolytics and antidepressants. OBJECTIVES: The aim of this review is to assess the effectiveness of anxiolytic drugs in aiding long term smoking cessation. The drugs include buspirone; diazepam; doxepin; meprobamate; ondansetron; and the beta-blockers metoprolol, oxprenolol and propanolol. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register which includes trials indexed in Medline, Embase, SciSearch and PsycLit, and meetings abstracts. SELECTION CRITERIA: We considered randomized trials comparing anxiolytic drugs to placebo or an alternative therapeutic control for smoking cessation. We excluded trials with less than 6 months follow-up. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the type of study population, the nature of the drug therapy, the outcome measures, method of randomisation, and completeness of follow-up. The main outcome measure was abstinence from smoking after at least six months follow-up in patients smoking at baseline. We used the most rigorous definition of abstinence for each trial, and biochemically validated rates if available. Where appropriate, we performed meta-analysis using a fixed effects model. MAIN RESULTS: There was one trial each of the anxiolytics diazepam, meprobamate, metoprolol and oxprenolol. There were two trials of the anxiolytic buspirone. None of the trials showed strong evidence of an effect for any of these drugs in helping smokers to quit. However, confidence intervals were wide, and an effect of anxiolytics cannot be ruled out on current evidence. REVIEWER'S CONCLUSIONS: There is no consistent evidence that anxiolytics aid smoking cessation, but the available evidence does not rule out a possible effect. PMID- 11034775 TI - Vitamin E for Alzheimer's disease. AB - BACKGROUND: Vitamin E is a dietary compound that functions as an antioxidant scavenging toxic free radicals. Evidence that free radicals may contribute to the pathological processes in Alzheimer's disease has led to interest in the use of vitamin E in the treatment of this disorder. OBJECTIVES: To examine the effects of vitamin E treatment for people with Alzheimer's disease. SEARCH STRATEGY: The Cochrane Dementia Group Register of Clinical Trials was searched with the following terms: vitamin E, Alzheimer's disease, dementia, alpha-tocopherol, cognitive impairment, cognitive function and controlled trials. The latest search was carried out in July 2000. SELECTION CRITERIA: All unconfounded, double blind, randomized trials in which treatment with vitamin E at any dose was compared with placebo for patients with Alzheimer's disease. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied the selection criteria an assessed study quality. One reviewer extracted and analysed the data. For each outcome measure data were sought on every patient randomized. Where such data were not available an analysis of patients who completed treatment was conducted. MAIN RESULTS: Only one study was identified which met the inclusion criteria (Sano 1997). The primary outcome used in this study of 341 participants was survival time to the first of 4 endpoints, death, institutionalisation, loss of 2 out of 3 basic activities of daily living, or severe dementia, defined as a global Clinical Dementia Rating of 3. The investigators reported the total numbers in each group who reached the primary endpoint within two years for participants completing the study ("completers"). There appeared to be some benefit from vitamin E with fewer participants reaching endpoint - 58% (45/77) of completers compared with 74% (58/78) - a Peto odds ratio of 0.49, 95% confidence interval 0.25 to 0.96. However, more participants taking vitamin E suffered a fall (12/77 compared with 4/78; odds ratio 3.07, 95% CI 1.09 to 8.62). It was not possible to interpret the reported results for specific endpoints or for secondary outcomes of cognition, dependence, behavioural disturbance and activities of daily living. REVIEWER'S CONCLUSIONS: There is insufficient evidence of efficacy of vitamin E in the treatment of people with with Alzheimer's disease. The one published trial of acceptable methodology (Sano 1997) was restricted to patients with moderate disease, and the published results are difficult to interpret. There is sufficient evidence of possible benefit to justify further studies. There was an excess of falls in the vitamin E group compared with placebo which requires further evaluation. PMID- 11034776 TI - Amniotomy alone for induction of labour. AB - BACKGROUND: Amniotomy (deliberate rupture of the membranes) is a simple procedure which can be used alone for induction of labour if the membranes are accessible, thus avoiding the need for pharmacological intervention. However, the time interval from amniotomy to established labour may not be acceptable to clinicians and women, and in a number of cases labour may not ensue. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of amniotomy alone for third trimester labour induction in women with a live fetus. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled trials register and bibliographies of relevant papers. SELECTION CRITERIA: The criteria for inclusion included the following: (1) clinical trials comparing amniotomy alone for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random or pseudo-random allocation to the treatment or control group; (3) ideally adequate allocation concealment (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. A strategy was developed to deal with the large volume and complexity of trial data relating to labour induction. This involved a two-stage method of data extraction. The initial data extraction was done centrally, and incorporated into the series of primary reviews arranged by methods of induction of labour. The data from the primary reviews will be incorporated into a series of secondary reviews, arranged by category of woman to reflect clinical scenarios. To avoid duplication of data in the primary reviews, the labour induction methods have been listed in a specific order, from one to 25. Each primary review includes comparisons between one of the methods (from two to 25) with only those methods above it on the list. This review includes comparisons between amniotomy alone (number 5 on the list) with only those methods above it on the list (no treatment / placebo; intravaginal prostaglandins; intracervical prostaglandins; and oxytocin alone). MAIN RESULTS: Two trials comprising 50 and 260 women respectively were eligible for inclusion in this review. No conclusions could be drawn from comparisons of amniotomy alone versus no intervention, and amniotomy alone versus oxytocin alone (small trial, only one pre-specified outcome reported). No trials compared amniotomy alone with intracervical prostaglandins. One trial compared amniotomy alone with a single dose of vaginal prostaglandins for women with a favourable cervix, and found a significant increase in the need for oxytocin augmentation in the amniotomy alone group (44% versus 15%; RR 2.85, 95% CI 1.82-4.46). This should be viewed with caution as this was the result of a single centre trial. Furthermore, secondary intervention occurred 4 hours after amniotomy, and this time interval may not have been appropriate. REVIEWER'S CONCLUSIONS: Data is lacking about the value of amniotomy alone for induction of labour. While there are now other modern methods available for induction of labour (pharmacological agents), there remain clinical scenarios where amniotomy alone may be desirable and appropriate, and this method is worthy of further research. This research should include evaluation of the appropriate time interval from amniotomy to secondary intervention, women and caregivers' satisfaction and economic analysis. PMID- 11034777 TI - Oral beta-blockers for mild to moderate hypertension during pregnancy. AB - BACKGROUND: Hypertension is a common complication of pregnancy. Antihypertensive drugs are widely used in the belief these will improve outcome for both the woman (such as decreasing the risk of stroke or eclampsia) and her baby (such as decreasing the risk of preterm birth and its complications). Beta-blockers are a popular choice of antihypertensive agent during pregnancy; other choices include methyldopa and calcium channel blockers. OBJECTIVES: The aim of this review is to assess whether oral beta-blockers are overall better than placebo, or no beta blocker, for women with mild-moderate hypertension during pregnancy, and to assess whether oral beta-blockers have any advantages over other antihypertensive agents for women with mild-moderate hypertension during pregnancy. Both maternal outcomes (e.g., the incidence of severe hypertension) and perinatal outcomes (e.g., mortality) were of interest. SEARCH STRATEGY: Register of trials maintained by the Cochrane Pregnancy and Childbirth Group, MEDLINE 1966-97, bibliographies of retrieved papers, personal files. Date of last search: June 2000. SELECTION CRITERIA: Trials comparing beta-blockers with (i) placebo or no therapy, or (ii) other antihypertensive agents, for women with mild-moderate pregnancy hypertension (i.e., blood pressure under 170 mm Hg systolic, or 110 mm Hg diastolic). DATA COLLECTION AND ANALYSIS: All data were extracted independently by two investigators, who were not blinded to outcome or other trial characteristics. Whenever possible, missing data were obtained by personal communication with authors. Discrepancies were resolved by discussion. The overview was divided into two comparisons: (i) beta-blockers versus placebo or no therapy, and (ii) beta-blockers versus other antihypertensives. MAIN RESULTS: Twenty-seven trials, involving just under 2400 women, are included in this review. Fourteen trials (1516 women) compared beta-blockers with placebo/no beta blocker. Oral beta-blockers decrease the risk of severe hypertension (relative risk (RR) 0.37, 95% confidence interval (CI) 0.26-0.53) and the need for additional antihypertensive drugs (RR 0.44, 95% CI 0.31-0.62). There are insufficient data for any conclusions about the effect on perinatal mortality or preterm delivery. Beta-blockers seem to be associated with an increase in small for gestational age infants (RR 1.34, 95% CI 1.01-1.79). Maternal hospital admission may be decreased, neonatal bradycardia increased and respiratory distress syndrome decreased, but these outcomes are only reported in a very small proportion of trials. Eleven trials (787 women) compared beta-blockers with methyldopa. Beta-blockers appear to be no more effective and probably equally as safe (from maternal and perinatal perspectives) as methyldopa. Single small trials have compared beta-blockers with hydralazine and with nicardipine. It is unusual for women to change drugs due to side effects. REVIEWER'S CONCLUSIONS: The improvement in control of maternal blood pressure with use of beta-blockers would be worthwhile only if it were reflected in other more substantive benefits for the mother and/or baby, and none have yet been clearly demonstrated. The effect of beta-blockers on perinatal outcome is uncertain, given that the worrying trend to an increase in small for gestational age infants is partly dependent on one small outlying trial. Large, randomised controlled trials are needed to determine whether antihypertensive therapy in general (rather than beta blocker therapy specifically) results in benefits that outweigh the risks for treatment of mild-moderate pregnancy hypertension. If so, then it would be appropriate to look at which antihypertensive is best. Beta-blockers would remain a candidate class of agents. PMID- 11034778 TI - Intravenous prostaglandin for induction of labour. AB - BACKGROUND: Intravenous prostaglandin E2 and F2 alpha can be used to induce labour. The use of intravenous prostaglandins in this context has been limited by perceived unacceptable maternal side effect profiles. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of intravenous prostaglandin for third trimester cervical ripening or induction of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, The Cochrane Controlled Trials Register and bibliographies of relevant papers. SELECTION CRITERIA: The criteria for inclusion included the following: (1) clinical trials comparing intravenous prostaglandin used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involves a two-stage method of data extraction. The initial data extraction is done centrally, and incorporated into a series of primary reviews arranged by methods of induction of labour, following a standardised methodology. The data will then be extracted from the primary reviews into a series of secondary reviews, arranged by category of woman. To avoid duplication of data in the primary reviews, the labour induction methods have been listed in a specific order, from one to 25. Each primary review includes comparisons between one of the methods (from two to 25) with only those methods above it on the list. MAIN RESULTS: Thirteen trials were eligible for inclusion in this review. Two trials (comprising 400 women) compared intravenous prostaglandin E2 to intravenous oxytocin, a further seven trials (comprising 590 women) compared intravenous prostaglandin F2 alpha to intravenous oxytocin. Two trials (comprising 115 women) each randomised women to one of three treatment arms namely intravenous oxytocin or intravenous prostaglandin F2 alpha or prostaglandin E2. One trial reported a comparison of combined oxytocin and prostaglandin F2 alpha and oxytocin alone in 20 women and lastly one trial compared extra amniotic prostaglandin E2 versus intravenous prostaglandin E2 (40 women). The use of intravenous prostaglandin was associated with higher rates of uterine hyperstimulation with changes in the fetal heart rate (relative risk (RR) 6.76, 95% confidence interval (CI) 1.23-37.11) and without (RR 4.25, 95%CI 1.48 12.24) compared to oxytocin. Use of prostaglandins was also associated with significantly more maternal side effects (gastrointestinal, thrombophlebitis and pyrexia, RR 3.75, 95% CI 2.46-5.70) than oxytocin. Prostaglandin was no more likely to result in vaginal delivery than oxytocin (RR 0.85, 95% CI 0.61-1.18). No significant differences emerged from subgroup analysis or from the trials comparing combination oxytocin/prostaglandin F2 alpha and oxytocin or extra amniotic versus intravenous prostaglandin E2. REVIEWER'S CONCLUSIONS: Intravenous prostaglandin is no more efficient than intravenous oxytocin for the induction of labour but its use is associated with higher rates of maternal side effects and uterine hyperstimulation than oxytocin. No conclusions can be drawn form the comparisons of combination of prostaglandin F2 alpha and oxytocin compared to oxytocin alone or extra amniotic and intravenous prostaglandin E2. PMID- 11034779 TI - Mifepristone for induction of labour. AB - BACKGROUND: The steroid hormone, progesterone, inhibits contractions of the uterus. Antiprogestins (including mifepristone) have been developed to antagonise the action of progesterone, and these have a recognised role in medical termination of early or mid-pregnancy. Animal studies have suggested that mifepristone may also have a role in inducing labour in late pregnancy. OBJECTIVES: To determine the effects of mifepristone for third trimester cervical ripening or induction of labour. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register, and reference lists of relevant papers. SELECTION CRITERIA: Selection criteria included: (1) clinical trials comparing mifepristone used for third trimester cervical ripening or labour induction with placebo/no treatment or other labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. Data were extracted by the reviewer and, independently, by a colleague. MAIN RESULTS: Seven trials, that recruited 594 women, are included. All trials compared mifepristone with placebo, except for one that compared mifepristone with no treatment. Compared to placebo, mifepristone treated women were less likely to have an unfavourable cervix at 48 hours (relative risk [RR] 0.36, 95% confidence intervals [CI] 0.2-0.63) or 96 hours (RR 0.39, 95% CI 0.23-0.66). Mifepristone treated women were more likely to have delivered within 48 and 96 hours of treatment than were placebo treated/no treatment women - 48 hours: RR 2.82, 95% CI 1.82-4.36; 96 hours: RR 3.40, 95% CI 1.96-5.92. Mifepristone treated women were less likely to undergo caesarean section (RR 0.71, 95% CI 0.53-0.95). There is little information about fetal outcome, although there was no evidence that neonatal hypoglycaemia might be more common after exposure to mifepristone. Similarly, there is little information about maternal side-effects although some nausea and vomiting was reported in one trial. REVIEWER'S CONCLUSIONS: There is insufficient information available from clinical trials to support the use of mifepristone to induce labour. However, available data do show that mifepristone is better than placebo at ripening the cervix, and inducing labour. There is evidence of a possible reduction in the incidence of caesarean section following mifepristone treatment (compared to placebo) that would justify further trials. We found no trials that compared mifepristone with alternative methods of inducing labour e.g. prostaglandins. PMID- 11034780 TI - Individual or group antenatal education for childbirth/parenthood. AB - BACKGROUND: Structured antenatal education programs for childbirth and/or parenthood are commonly recommended for pregnant women and their partners by health care professionals in many parts of the world. Such programs are usually offered to groups but may be offered to individuals. OBJECTIVES: The objective of this review was to assess the effects of this education on knowledge acquisition, anxiety, sense of control, pain, support, breastfeeding, infant care abilities, and psychological and social adjustment. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register, and other databases were searched. The date of the last search was December, 1999. SELECTION CRITERIA: Randomized controlled trials of any structured educational program provided during pregnancy by an educator to either parent, that included information related to pregnancy, birth, or parenthood were included. The educational interventions could have been provided on an individual or group basis. Educational interventions directed exclusively to either increasing breastfeeding success or reducing smoking were excluded, since reviews of these topics can be found elsewhere in The Cochrane Library. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data were extracted by the author from published reports. MAIN RESULTS: Six trials, involving 1443 women, were included. Twenty-two were excluded. The largest of the included studies (n = 1275) examined an educational intervention to increase vaginal birth after cesarean section. This high quality study showed similar rates of vaginal birth after cesarean section in 'verbal' and 'document' groups, relative risk (RR) 1.1 (95% confidence interval (CI) 1.0-1.2). More general educational interventions were the focus of the other five studies (combined n = 168). The methodological quality of these trials is uncertain, since details of the randomization procedure, allocation concealment, and/or participant accrual/loss were not reported. No consistent results were found. Sample sizes were very small, ranging from 10-67. Interventions, populations, and outcomes measured were different in each study. No data from the five general education trials were reported concerning labour and birth outcomes, anxiety, breastfeeding success, or general social support. Knowledge acquisition and factors related to infant care competencies were measured. REVIEWER'S CONCLUSIONS: Individualized prenatal education directed toward avoidance of a cesarean birth does not increase the rate of vaginal birth after cesarean section. The effects of general antenatal education for childbirth and/or parenthood remain unknown. PMID- 11034781 TI - A Secondary beta Deuterium Kinetic Isotope Effect in the Chorismate Synthase Reaction. AB - Chorismate synthase (EC 4.6.1.4) is the shikimate pathway enzyme that catalyzes the conversion of 5-enolpyruvylshikimate 3-phosphate (EPSP) to chorismate. The enzyme reaction is unusual because it involves a trans-1,4 elimination of the C-3 phosphate and the C-6 proR hydrogen and it has an absolute requirement for reduced flavin. Several mechanisms have been proposed to account for the cofactor requirement and stereochemistry of the reaction, including a radical mechanism. This paper describes the synthesis of [4-(2)H]EPSP and the observation of kinetic isotope effects using this substrate with both Neurospora crassa and Escherichia coli chorismate synthases. The magnitude of the effects were (D)(V) = 1.08 +/- 0.01 for the N. crassa enzyme and 1.10 +/- 0.02 on phosphate release under single turnover conditions for the E. coli enzyme. The effects are best rationalised as substantial secondary beta isotope effects. It is most likely that the C(3)-O bond is cleaved first in a nonconcerted E1 or radical reaction mechanism. Although this study alone cannot rule out a concerted E2-type mechanism, the C(3) O bond would have to be substantially more broken than the proR C(6)-H bond in a transition state of such a mechanism. Importantly, although the E. coli and N. crassa enzymes have different rate limiting steps, their catalytic mechanisms are most likely to be chemically identical. Copyright 2000 Academic Press. PMID- 11034782 TI - A Structure-Activity Relationship for the Hydrolysis of Acetylamino Acids by Porcine Aminoacylase. AB - A structure-activity relationship is presented that satisfactorily predicts the rates of hydrolysis of a series of acetylglycine derivatives by porcine aminoacylase. It is apparent that the substrate specificity of aminoacylase is mainly kinetic in origin, the observed correlation with Taft's E(s) parameter supporting the notion that enzymolysis proceeds through a mechanism that is analogous to chemical hydrolysis. It is suggested that the alpha-CH(2)CH group of those substrates that possess this moiety is conformationally immobile upon binding. This lock facilitates rapid hydrolysis and results from steric interactions between the enzyme and substrate. The incorporation of alpha-methyl amino acid derivatives in the structure-activity relationship is consistent with a flexible active site model and it is concluded that the alpha-methyl effect in this system is a binding phenomenon. It is evident that the active center of porcine aminoacylase can comfortably accommodate amino acid derivatives with side chains containing less than six carbon atoms, contrary to previous assertions. It is suggested that the binding of bulkier derivatives necessitates the distortion of the active site. Derivatives possessing beta-hydroxyl groups are found to deviate from expected behavior and a nonproductive binding model is presented. Copyright 2000 Academic Press. PMID- 11034783 TI - Antimicrobial and Biological Effects of Bomphos and Phomphos on Bacterial and Yeast Cells. AB - In this study, the antimicrobial effects of monophosphazenes such as SM, BOMPHOS, and PHOMPHOS were examined on bacterial and yeast strains. In addition, the biological effects of these compounds were tested on the Saccharomyces cerevisiae and Candida albicans cells. The SM has an antimicrobial effect on the bacterial and yeast strains within the range of 100 and 1500 ug. When the concentration was increased, the inhibition zone expanded on the growth media (P < 0.01; P < 0.001). Like SM, BOMPHOS molecule has antimicrobial activity on the bacterial and yeast cells. The most effective concentrations of BOMPHOS on the microorganisms were observed by 1500 ug (P < 0.001). The PHOMPHOS did not effect on the bacterial and yeast cells between 100 and 1000 range, but it has an antimicrobial effect in 1500 ug. In vitro media, the biological effects of these molecules were compared with vitamin E, melatonin, and fish oil on the yeast cells. In S. cerevisiae growth media, the cell densities were increased SM, BOMPHOS, and PHOMPHOS after 20, 30, and 45 h. The highest increase in the cell density were observed in media of BOMPHOS. In C. albicans growth media, the cell density was increased by melatonin after 20, 30, and 45 h, but were decreased by other supplemental groups. Lipid level of S. cerevisiae was reduced by administered 300 and 1000 ug vitamin E and fish oil (P < 0.01). In addition, the lipid level of the same yeast cell were diminished by the 1000 ug melatonin and 300 ug PHOMPHOS (P < 0.05, P < 0.01). The lipid level of C. albicans were increased by vitamin E and BOMPHOS and fish oil, but was decreased with PHOMPHOS (P < 0.01). In conclusion, while high concentration of PHOMPHOS has antimicrobial effects on the bacterial and yeast cells, the SM and BOMPHOS have antimicrobial effects in all the concentrations. PHOMPHOS decreased the lipid level of C. albicans, but BOMPHOS increased in the the same yeast cell. In addition, the antioxidants such as vitamin E, melatonin, and fish oils have affected on the lipid synthesis of yeast cells. Copyright 2000 Academic Press. PMID- 11034784 TI - Inhibition Effects in the Hydrolysis Reactions of Esters and Peptides Catalyzed by Carboxypeptidase A: An Example of Cooperative Binding Effects with a Monomeric Enzyme. AB - N-benzoyl-L-phenylalanyl-L-phenylalanine is an excellent peptide substrate for carboxy-peptidase A; at 30 degrees C and pH 7.5, K(m) is 2.6 x 10(-5) M while k(cat) is 177 s(-1) (k(cat)/K(m) = 6.8 x 10(6) M(-1) s(-1)). Indole-3-acetic acid is a noncompetitive or mixed inhibitor towards the peptide and toward hippuryl-L phenylalanine; plots of E/V vs [Inhibitor] are linear. N-Benzoyl-L-phenylalanine is a competitive inhibitor of peptide hydrolysis, and plots of E/V vs [Inhibitor] are again linear. One molecule of inhibitor binds per active site, and these inhibitors bind in different sites. At constant peptide substrate concentration and a series of constant concentrations of indole-3-acetic acid, plots of E/V vs the concentration of N-benzoyl-L-phenylalanine are linear and intersect behind the E/V axis and above the [Inhibitor] axis. This shows that both inhibitors can bind simultaneously and that binding of one facilitates the binding of the other (beta = 0.18). Employing the ester substrate hippuryl-DL,beta-phenyllactate, the same type of behavior is observed in the reverse sense; N-benzoyl-L-phenylalanine is a linear noncompetitive inhibitor and indole-3-acetic acid is a linear competitive inhibitor. Again the two inhibitor plot is linear and intersects above the [Inhibitor] axis (beta = 0.12). Previous X-ray crystallographic studies have indicated that indole-3-acetic acid binds in the hydrophobic pocket of the S'(1) site, while N-benzoyl-L-phenylalanine binds in the S(1)-S(2) site. The product complex for hydrolysis of N-benzoyl-L-phenylalanyl-L-phenylalanine (phenylalanine + N-benzoyl-L-phenylalanine) occupies both of these sites. However, the present work shows that the peptide substrate does not bind to the enzyme at pH 7.5 so as to be competitive with indole-3-acetic acid. The binding sites may be formed via conformational changes induced or stabilized by substrate and product binding. Copyright 2000 Academic Press. PMID- 11034785 TI - Distinct Effect of Benzalkonium Chloride on the Esterase and Aryl Acylamidase Activities of Butyrylcholinesterase. AB - Acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) from vertebrates, other than their predominant acylcholine hydrolase (esterase) activity, display a genuine aryl acylamidase activity (AAA) capable of hydrolyzing the synthetic substrate o-nitroacetanilide to o-nitroaniline. This AAA activity is strongly inhibited by classical cholinesterase (ChE) inhibitors. In the present study, benzalkonium chloride (BAC), a cationic detergent widely used as a preservative in pharmaceutical preparations, has been shown to distinctly modulate the esterase and AAA activities of BChEs. The detergent BAC was able to inhibit the esterase activity of human serum and horse serum BChEs and AChEs from electric eel and human erythrocyte. The remarkable property of BAC was its ability to profoundly activate the AAA activity of human serum and horse serum BChEs but not the AAA activity of AChEs. Thus BAC seem to preferentially activate the AAA activity of BChEs alone. Results of the study using the ChE active site-specific inhibitor diisopropyl phosphorofluoridate indicated that BAC binds to the active site of ChEs. Furthermore, studies using a structural homolog of BAC indicated that the alkyl group of BAC is essential not only for its interaction with ChEs but also for its distinct effect on the esterase and AAA activities of BChEs. This is the first report of a compound that inhibits the esterase activity, while simultaneously activating the AAA activity, of BChEs. Copyright 2000 Academic Press. PMID- 11034786 TI - Differences Arising in Human Neutrophil Activation Passing from N-Formyl to N Acetyl-oligopeptides. AB - N-formyl- and N-acetyl-peptides were synthesized and compared in order to understand which features can best elicit biological responses. The behavior of N formyl-peptides confirms the previously found sequential obligations in the residues, while acetyl-derivatives do not seem suitable for an efficacious stimulation of human neutrophils. Copyright 2000 Academic Press. PMID- 11034787 TI - Variation of the HES concentration for the cryopreservation of keratinocytes in suspensions and in monolayers. AB - It has recently been shown that keratinocytes, both in suspension and in monolayers, can be successfully cryopreserved with hydroxyethyl starch (HES) (6, 9). HES is a nontoxic biodegradable macromolecule which is clinically approved as a plasma expander and which has already been used for the cryopreservation of red blood cells (10, 11). In this study we varied the HES concentration between 0 and 10 wt% in 2% steps for suspended cells and between 0, 4, 6, 8, and 10 wt% for monolayer cells in order to determine the effect on the survival rate and metabolic activity after cryopreservation. The experiments with the suspended cells were performed both with and without NCS. Cryopreserved keratinocytes can be transplanted onto patients for the treatment of deep dermal burns and leg ulcers. In this study, we achieved a survival rate of 80% for the suspended cells (10 wt% HES, 3 degrees C/min) and a survival rate of even 88% when the cells were cryopreserved as a monolayer using the same parameters. The addition of NCS did not improve the results for the suspended cells significantly. PMID- 11034788 TI - Platelet cryopreservation using a combination of epinephrine and dimethyl sulfoxide as cryoprotectants. AB - Current methods of platelet storage are unsatisfactory because of the short shelf life of platelets and the rapid loss of platelet viability. We have developed a cryopreservation method that results in less damage from freezing and higher recovered function of platelets. Platelets were cryopreserved using a combination of epinephrine (EPN) and dimethyl sulfoxide (Me(2)SO) as cryoprotectants. The response of platelets to agonists was studied by flow cytometry and aggregation tests. Cryopreserving platelets with Me(2)SO decreased platelet annexin V binding due to freezing. The combination of EPN with Me(2)SO enhanced Me(2)SO cryoprotection and decreased platelet microparticle generation, suggesting that cryopreserving platelets using this combination is associated with increased platelet integrity. Platelet cryopreservation with an Me(2)SO/EPN combination also increased platelet aggregability, which was demonstrated by decreasing the lag phase and increasing the aggregation density to 66.39% +/- 6.6 that of fresh platelet-rich plasmas. We conclude that adding EPN as a combined cryoprotectant improves the quality of Me(2)SO-frozen platelets. As a method of aggregation of cryopreserved platelets, this method is comparable to that of normal fresh platelets and may improve the conditions for platelet transfusion. PMID- 11034789 TI - Osmotic behavior of in vitro produced bovine blastocysts in cryoprotectant solutions as a potential predictive test of survival. AB - The osmotic behavior of bovine blastocysts produced in vitro was filmed during exposure to and dilution of cryoprotectant solutions used for vitrification. The relationship between the changes in the diameter of embryos and their subsequent survival was assessed. Embryos collected on Day 6 and Day 7 postinsemination were exposed to 10% glycerol (GLY) for 5 min, 10% GLY + 20% ethylene glycol (EG) for 5 min, and 25% Gly + 25% EG for 30 s, before dilution in 0.85 M galactose and finally in embryo transfer freezing medium (ETF). Embryos that had a higher probability of survival behaved as perfect osmometers, shrinking, reexpanding, or swelling according to an identical pattern, whereas embryos that deviated from this standard usually did not survive. The initial embryo diameter, duration of shrinkage and expansion in 10% glycerol, duration of reexpansion in ETF, and final embryo diameter were clearly predictive of the ability to hatch after culture in vitro. On a given day postinsemination, larger blastocysts were more likely than smaller blastocysts to survive and hatch after exposure to cryoprotectants with or without vitrification. PMID- 11034790 TI - Piglets born after vitrification of embryos using the open pulled straw method. AB - Morulae and unhatched blastocysts from Large White hyperprolific (LWh) and Meishan (MS) gilts were selected to test an ultrarapid open pulled straw (OPS) vitrification method with two media. The viability of vitrified/warmed embryos was estimated by the percentage of embryos that developed to the hatched blastocyst stage in vitro or by birth after transfer. In Experiment 1, two cryoprotectant dilution media were compared for cryopreservation of MS and LWh blastocysts: TCM was a standard Hepes-buffered TCM199 + 20% NBCS medium and PBS was a PBS + 20% NBCS medium. After a two-step equilibration in ethylene glycol, dimethyl sulfoxide, and sucrose, 2-5 blastocysts were loaded into OPS and plunged into liquid nitrogen. Embryos were warmed; a four-step dilution with decreasing concentrations of sucrose was applied. In PBS, LWh blastocysts (27%) had a lower viability in vitro than MS blastocysts (67%; P = 0.001). In TCM, no significant difference was observed between genotypes (41% for LWh and 43% for MS blastocysts) and both viability rates were lower than that of the control groups. In Experiment 2, morula-stage LWh and MS embryos were vitrified and warmed using PBS. The viability rate was low and did not differ between LWh (11%) and MS (14%). In Experiment 3, 200 MS and 200 LWh blastocysts were vitrified/warmed as described in Experiment 1 (PBS). In each of 20 MS recipients, 20 embryos were transferred. The farrowing rate was 55% and recipients farrowed four and five piglets (median) for MS and LWh blastocysts, respectively. The OPS method is therefore appropriate for cryopreservation of unhatched porcine blastocysts. PMID- 11034791 TI - Effect of cooling on the motility and function of human spermatozoa. AB - Human spermatozoa were cooled from 37 to 0 degrees C at 10 degrees C min(-1) in 5 degrees C steps with 1 min equilibration at each step, the temperature control was +/- 0.1 degrees C. Spermatozoa were held at 0 degrees C for 5 min and then rewarmed at the same rate. No significant effect of cooling on the straight-line velocity was found using computer-aided semen analysis. The physiological function of spermatozoa was also examined before and after cooling using hypoosmotic swelling, ionophore-provoked acrosome reaction, and binding to fragments of human zonae pellucidae. Spermatozoa were cooled either in seminal plasma or in conventional IVF medium with or without fractionation by centrifugation through a discontinuous Percoll gradient. When spermatozoa were cooled and rewarmed in seminal plasma there was no significant change in either the ionophore-induced acrosome reaction or the binding to zona pellucida fragments. When spermatozoa were fractionated by centrifugation through Percoll an increased response in both was seen. However, following cooling and rewarming, a significant decline in the response of both occurred. We suggest that motility alone is not a reliable predictor of changes in other physiological functions of spermatozoa following cooling. Furthermore, short-term cooling appears to have no significant detrimental effect on normozoospermic samples and cold shock may be avoided in the clinical context by controlled cooling and warming. PMID- 11034792 TI - Cryopreserved rat liver slices: a critical evaluation of cell viability, histological integrity, and drug-metabolizing enzymes. AB - The effects of a cryopreservation procedure on the biochemical, morphological and functional integrity of rat liver slices just after thawing and after 24 h culture were evaluated. Freshly prepared slices were incubated in modified University of Wisconsin solution containing 50% fetal calf serum and 10% dimethyl sulfoxide for 20 min on ice prior to a rapid cooling in liquid nitrogen. After 10 40 days, slices were thawed rapidly at 42 degrees C. Total protein content and (3 [4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) (MTT) reduction were well preserved at thawing, whereas ATP content was markedly decreased relative to freshly prepared slices (-83%). The major microscopic findings in sections of just-thawed liver slices consisted of hepatocellular dissociation and minimal apoptosis. The qualitative profile of antipyrine (AP) metabolism was well preserved in cryopreserved slices, but the amounts of phase I and phase II AP metabolites produced over a 3-h incubation period were markedly reduced relative to fresh slices (-58 to -71%). When cryopreserved slices were cultured for 24 h after thawing, the viability was markedly reduced, as reflected by the almost complete absence of MTT reduction and the loss of ATP content. Histological examinations showed extensive cellular necrosis. The amount of AP metabolites produced by cryopreserved slices was similar after a 3- or a 24-h culture period, indicating that AP metabolism capacities were lost at 24 h culture. In conclusion, our results suggest that cryopreserved rat liver slices may be a useful model for short-term in vitro determination of drug metabolism pathways. Further work is required to extend their use for toxicological studies. PMID- 11034793 TI - Bile analysis as a tool for assessing integrity of biliary epithelial cells after cold ischemia--reperfusion of rat livers. AB - Previous morphological studies failed to show appreciable injury of biliary epithelial cells (BEC) after cold ischemia of rat liver, although recent evidence indicated that BEC integrity and function were impaired in this model. We tested the hypothesis that analysis of bile for enzymes, such as lactate dehydrogenase (LDH), alanine transaminase (ALT), and aspartate transaminase (AST), can be used for assessing cold ischemic injury of BEC. Furthermore, we examined whether biliary gamma-glutamyltransferase (GGT) reflects warm ischemic injury of BEC and whether normothermic reperfusion aggravates the negative effect of cold ischemia on BEC integrity and function. Rat livers were reperfused after different periods of cold or warm ischemia using a blood-free perfusion model. Compared with controls, perfusate LDH, ALT, and AST levels and parameters of hepatocyte function, including hepatocyte tight junction permeability, were not significantly altered by 18-h cold ischemia. On the other hand, 9-h cold ischemia markedly increased biliary LDH, ALT, and AST levels. However, only LDH release into the bile was strongly dependent on the time of cold storage. Biliary GGT, LDH, and glucose levels decreased during the reperfusion period following 18-h cold ischemia. The results suggest that biliary LDH can be used for assessing injury of BEC in cold-preserved livers and that normothermic reperfusion does not aggravate preservation-induced injury of BEC after cold ischemic storage. PMID- 11034794 TI - Cryopreservation of Musca domestica (Diptera: Muscidae) embryos. AB - Prior studies on cryopreserving embryos of several non-drosophilid flies established that two Drosophila melanogaster embryo cryopreservation protocols were not directly suitable for use with these species. This paper describes our work on developing a protocol for cryopreservation of embryos of the housefly, Musca domestica. Significant progress was made when permeabilization of the vitelline membrane was optimized, a vitrification solution containing ethylene glycol, polyethylene glycol, and trehalose was formulated, and when cooling and recovery of the cryopreservation protocol included a step which passed the embryos through liquid nitrogen vapor. More than 70% of housefly embryos withstand treatments of dechorionation, permeabilization, loading with cryoprotectant, and dehydration in vitrification solution, but the cooling, warming, and poststorage rearing steps still cause a considerable reduction in survival. About 53% of the vitrified M. domestica embryos hatched into larvae. Relative to the percentage of the control adult emergence, about 13% of the embryos stored in liquid nitrogen developed into fertile adults. Hatching of the F(1) progeny of adults having been cryopreserved as embryos was similar to control levels. PMID- 11034795 TI - Measurement of the water permeability of the membranes of boar, ram, and rabbit spermatozoa using concentration-dependent self-quenching of an entrapped fluorophore. AB - Published values for sperm membrane water permeability (L(p)) obtained using a time-to-lysis methodology have produced anomalous results when used to model optimal cooling rates for cryopreservation of spermatozoa. As the lysis method is dependent on potentially questionable assumptions, we describe an alternative method for measuring sperm L(p). Spermatozoa were exposed to hypo- and hyperosmotic conditions using a stopped-flow apparatus and the time course of resulting volume changes was measured using concentration-dependent self quenching of the entrapped fluorophore, carboxyfluorescein (CF). L(p) was measured for boar, rabbit, and ram spermatozoa using a range of osmotic stresses (+/-50-100 mOsm). Values for exosmotic and endosmotic flow showed no evidence of rectification. Mean L(p) values were 0.84 microm/min/atm (boar), 0.28 microm/min/atm (rabbit), and 2.79 microm/min/atm (ram). These values are lower than the lysis method estimates, with the ram value reduced by approximately two thirds using the current methodology. The value for boar spermatozoa showed good agreement with published values obtained using an electronic cell-sizing technique. Substitution of the revised values for L(p) into the model for optimal cooling rates brings the calculated optimal rate closer to the lower empirically observed value but does not fully account for the previously reported discrepancies. PMID- 11034796 TI - EDITORIAL: The human genome project. PMID- 11034797 TI - COMMENTARY. AB - Commentary regarding: F. Cavaliere et al.-Peritonectomy and hyperthermic antiblastic perfusion in the treatment of peritoneal carcinomatosis. Copyright 2000 Harcourt Publishers Ltd. PMID- 11034798 TI - Educational tips in the treatment of malignant ulcerating tumours of the skin. AB - Coping with ulcerating or bleeding tumours or metastases of the skin that are not suitable for curative or palliative treatment poses a problem for patients, doctors and nursing staff. Treatment should focus on limiting local and systemic infection, combating unpleasant odours and reducing bleeding. Palliative amputation is sometimes a reasonable option. Treatment depends on the nature and site of the tumour and should be tailored to and carried out in consultation with the patient, the treating specialist and the oncology nursing team. The goal of treatment is to optimize the quality of life of patients in the terminal phase. PMID- 11034799 TI - Reply to Professor Bozzetti. PMID- 11034800 TI - Breast substitution. PMID- 11034801 TI - Detection of circulating breast cancer cells by reverse transcriptase polymerase chain reaction (RT-PCR). AB - The confounding problem in treatment of breast cancer is the metastasis of breast tumour. Reverse transcriptase polymerase chain reaction (RT-PCR) has been recently used in the detection of circulating breast cancer cells. This review reports on the development of this assay as well as its advantages and disadvantages. We feel that cytokeratin 20 and beta -human chorionic gonadotropin (hCG) mRNA are the best markers for the detection of circulating breast cancer cells. We suggest that the multiple RNA marker RT-PCR assay can help to increase both sensitivity and specificity of detection, and that quantitative RT-PCR assay is more effective than the qualitative assay in the detection of circulating breast cancer cells. PMID- 11034802 TI - Has the breast cancer 'two week wait' guarantee for assessment made any difference? AB - AIMS: To determine the referral practice of GPs to a Fast Access Breast Clinic before and after the implementation of the 'two week wait' and to demonstrate the impact of this guarantee on the detection rate of breast cancer and access to the Breast Clinic. METHODS: A complete audit cycle was performed in a District General hospital (Eastbourne District General Hospital). The main outcome measures were detected breast cancer, clinical accuracy of the GPs and the waiting time for a Fast Access breast clinic. RESULTS: Prior to the implementation of the Government's Directive, the detection rate of breast cancer was 22% and all cancers were seen within 1 week of referral letter. The routine waiting time for non-urgent assessment was 4 weeks. The clinical accuracy of GP referrals was 42%. On the basis of the findings in the first part of the audit, guidelines were sent to the GPs to aid in their referrals to FABC, prior to the 1 April deadline. After the implementation the re-audit showed that the cancer detection rate had dropped to 19% and only 85% of patients were seen in 2 weeks. The routine wait for non-urgent assessment had gone up to a minimum of 8 weeks. The clinical accuracy had slightly increased to 49%. CONCLUSION: With the transfer of power on deciding the urgency of referrals from the specialist to the GP, there has been a decline in the cancer detection rate and an increase in the waiting time for patients not deemed urgent by the GPs. The decision on the urgency of referral should be in the hands of the relevant specialist who individually can decide the best way to run their Fast Access Breast Clinics and thus achieve the government's guarantee of a maximum two week wait for patients suspected of having breast cancer. PMID- 11034803 TI - Prolactin as a local growth promoter in patients with breast cancer: GCRI experience. AB - AIMS: The aim of this study was to evaluate the prognostic value of pre-operative prolactin (PRL) in conjunction with established prognosticators, and the risk of disease relapse in patients with early and advanced breast cancer. To confirm the hypothesis that PRL is produced by breast tumours molecular analysis of PRL, using immunohistochemistry, mRNA by RT-PCR and direct sequencing, was performed. Furthermore, presence of prolactin receptors (PRLR) was evaluated by immunohistochemical localization in these patients. METHODS: In 111 breast cancer patients, pre-operative PRL was determined by an immunoradiometric assay (IRMA) method. Immunohistochemical localization of PRL (IHL-PRL) and PRLR was performed on formalin-fixed, paraffin-embedded tissue sections. Expression of PRL mRNA was carried out by reverse transcriptase polymerase chain reaction (RT-PCR). RT-PCR PRL amplimer was sequenced and compared with human pituitary PRL amplimer. RESULTS: Fifty-eight per cent (64/111) of the patients had hyperprolactinaemia (PRL520.0 ng/ml). With increasing tumour size, a higher incidence of hyperprolactinaemia was noted which was statistically significant (r=0.34, P=0.0001). In stage III patients, and in node positive patients, the incidence of hyperprolactinaemia was significantly higher compared to their respective counterparts (stage II vs stage III, r=0.37, P=0.00006; node negative vs node positive, r=0.30, P=0.001). Hyperprolactinaemic patients had a significantly higher risk of developing recurrent/metastatic disease and a higher mortality risk as compared to patients with PRL <20.0 ng/ml. The multivariate survival analysis indicated that apart from disease stage, prognosis of patients with pre operative hyperprolactinaemia was poorer than that of patients with PRL <20.0 ng/ml. Seventy-eight per cent (87/111) of the tumours showed positive immunoreactivity with PRL antibody indicating that PRL, or a similar molecule, is produced ectopically by breast tumours. PRL mRNA expression using RT-PCR confirmed the de novo synthesis of PRL. PRL mRNA expression was seen in 52% (33/63) of tumours. Sequence analysis of the 234 bp PRL amplimer revealed that the sequence was homologous to the sequence of exon 5 of human pituitary PRL mRNA. Furthermore, PRLR were present in 80% of tumours detected by immunohistochemical localization. A significant positive correlation was noted between IHL-PRL and PRLR (r=0.26, P=0.006). CONCLUSIONS: This multifaceted study of PRL suggests that breast cancer cells produce PRL and that this ectopically produced PRL may act as a major local growth promoter via autocrine and paracrine mechanisms. It may provide new insights into endocrine treatment of breast cancer. PMID- 11034804 TI - Prognostic factors predicting survival in the treatment of retroperitoneal sarcoma. AB - METHODS: Retroperitoneal soft tissue sarcomas are rare tumours. The management of these tumours has been difficult because of low resectability and a high recurrence rate. A retrospective review of a prospectively compiled database of 32 consecutive patients with retroperitoneal sarcomas treated at Oulu University Hospital between 1977 and 1996 was performed. RESULTS: The resectability rate of primary tumours was 75%, and 44% of the patients underwent radical resection. The recurrence rate after radical resection was 57% and the resectability rate for recurrent tumours after radical primary operation, 50%. The actuarial overall 5 year survival rate was 31%, 10-year survival rate 19% and median survival 36 months. In univariate analysis the principal factors associated with prognosis were radical resection, recurrent disease, pre-operative loss of weight and histological tumour grade. Complete excision of the primary tumour was the only significant predictor of survival in multivariate analysis. CONCLUSIONS: Complete resection of retroperitoneal sarcoma continues to be the most important prognostic factor. The inefficiency of adjuvant therapy, the high recurrence rate and the very low chance of curing the patient after recurrence make the prognosis of these patients poor. PMID- 11034805 TI - Advanced squamous cell carcinoma of the base of the tongue treated with surgery and post-operative radiotherapy. AB - BACKGROUND: Early stage squamous cell carcinoma of the base of the tongue has been successfully treated with radiotherapy and brachytherapy. However, the vast majority of these tumours seen in Western Europe are already at an advanced stage. Medical records of 79 patients with squamous cell carcinoma of the base of the tongue treated between 1980 and 1994 were examined. METHODS: Eighty-three per cent of the primary tumours were stage T3 or T4. Fifty-nine patients were treated with surgery and post-operative radiotherapy. Quality of life assessment amongst the survivors was performed by means of a questionnaire. RESULTS: Five year disease free survival in patients undergoing excision for T3-T4 tumours was 59%. Patients with T2-T3 tumours undergoing partial excision of the tongue base had a 3 year recurrence free survival rate of 68%. Distant metastasis occurred in 16%. Seventy-eight per cent of the patients judged their quality of life to be near normal. CONCLUSION: Surgery and post-operative radiotherapy offer a reasonably good survival in advanced carcinoma of the base of the tongue with preservation of quality of life. PMID- 11034806 TI - Limited resection for carcinoma of the upper thoracic oesophagus is not a realistic option. AB - AIMS: Due to its anatomical position, carcinoma of the proximal oesophagus results in early invasion of adjoining structures, often precluding (radical) resection. We performed a retrospective study to compare the potentially curative and palliative treatment results in patients with proximal (i.e. at or above the carina) vs distal oesophageal carcinoma. METHODS: Over a 3-year period 30 patients with proximal and 145 patients with distal oesophageal cancer underwent surgery. RESULTS: Microscopically radical resection was achieved in 11/30 patients (43%) with a proximal tumour and in 96/145 patients (66%) with a distal tumour (P=0.007). Three-year survival was 13.8%vs 44.3% respectively; localization was an independent prognostic factor. Recurrent upper aero-digestive tract symptoms developed in 38% of the patients with a proximal tumour and in 19% of the patients with distal carcinoma (P<0.05). DISCUSSION: Carcinoma of the proximal oesophagus has a worse prognosis than more distal carcinomas. Definite cure is exceptional; many patients are ineffectively palliated. In patients with proximal oesophageal carcinoma surgery should not be performed outside clinical trials testing multimodality treatment. PMID- 11034807 TI - SIOP treatment guidelines for renal tumours in small infants: fact or fantasy? AB - AIMS: Since as far back as 1980, SIOP (Societe Internationale d>>Oncologie Pediatrique) have advocated primary nephrectomy (PN) only for unilateral renal tumours in patients > tumour (WT). Fourteen of the 25 patients (56%) were treated with PN, including four patients with CMN. In group B there was one patient (2%) with CMN and 40 patients with WT. Thirteen of the patients (31%) were treated with PN. A total of 15 patients were treated before 1980 and 26 after 1980. Eight of 15 (53%) patients were treated with PN before 1980 and 21/26 (81%) were pre-treated after 1980, according to the protocol. CONCLUSION: Despite the SIOP recommendations, only 56% of patients 1 year (9%). DISCUSSION: The number of parathyroid glands preserved in situ did not help predict the duration of post surgical calcium treatment, nor the final outcome of hypocalcaemia. However, when total calcium levels were compared in patients having had one or two glands preserved vs three or four parathyroid glands, it was possible to show that despite prolonged calcium administration, late calcaemias remained significantly lower during the first 6 months in patients with a smaller number of parathyroid glands. Hypoparathyroidism, defined functionally on the basis of requirement of calcium supplementation 1 year after surgery, occurred in 8.6% of patients after bilateral lobectomy (despite measurable but inappropriately low-PTH concentration). This outcome could have been predicted earlier (after 3 to 6 months) and the patients perhaps given the benefit of definitive vitamin D treatment earlier, in order to avoid late and prolonged hypocalcaemia. Evaluation after 1 year showed that only one patient out of 82 bilateral lobectomies (1.2%) had permanent hypoparathyroidism and needed calcium whereas hypocalcaemia was persistent in one out of four patients who had undergone a staged procedure (i.e. heterolateral lobectomy years after a previous operation). PMID- 11034809 TI - Neoadjuvant radio-chemotherapy in advanced primarilynon-resectable carcinomas of the pancreas. AB - AIM: To investigate the feasibility of neoadjuvant radio-chemotherapy (RCT) in the treatment of primarily non-resectable pancreas carcinoma the parameters tumour regression, possibility of subsequent resection and tolerability were examined. METHOD: Between 1995 and 1997, 27 patients with locally inoperable (assessed by CT criteria) pancreatic carcinoma received radio-chemotherapy for 5 weeks comprising irradiation (55.8 Gy) and chemotherapy with 5-fluorouracil (5 FU, 1000 mg/m(2)/day; 120 h continuous infusion) and mitomycin C (10 mg/m(2)i.v. bolus, day 2 and day 30) during the first and fifth week of radiotherapy. Two target volumes were irradiated with fractionated doses of 1.8 Gy up to a total of 50.4 Gy. Radiation was applied once a day five times a week and target volume 1 was irradiated with the same fractionated dose, and an additional boost of 5.4 Gy to make an overall total of 55.8 Gy. RESULTS: Sixteen patients underwent explorative laparotomy, 10 of these were resected (eight Whipple's procedures, two distal pancreatic resections), while six could not be resected due to peritoneal carcinosis (n=3), local irresectability (n=2) and liver cirrhosis (n=1). A further nine patients were found to have unresectable tumours on CT and did not undergo surgery after restaging (five of these patients were staged as <>, three patients had distant metastases and one patient refused surgery). In two patients RCT was abandoned because of progression of disease. CONCLUSIONS: The study protocol described is feasible without significant acute toxicity and when used the resectability rate was improved; the survival rate, however, was not improved. Additional intra-arterial or intraportal application of such drugs as mitomycin C or cisplatin may be necessary. PMID- 11034810 TI - Pre-clinical evaluation of the activity of gemcitabine as a basis for regional chemotherapy of pancreatic and colorectal cancer. AB - AIMS: To estimate the potential activity of gemcitabine for hepatic arterial infusion (HAI) chemotherapy in pancreatic and colorectal cancer. METHODS: The anti-proliferative effects of gemcitabine were determined in MIA PaCa-2 and PMH2/89 pancreatic and HT29 and NMG64/84 colon cancer cell lines and in fresh tumours from patients with liver metastases of colon, rectal and pancreatic cancer in vitro using the human tumour colony forming assay. RESULTS: Gemcitabine showed concentration and time-dependent cytotoxic effects in all tested cell lines. The IC(50)of gemcitabine in MIA PaCa-2, PMH2/89, HT29 and NMG64/84 cells at 2 h exposure time were >100, 18, 100 and 2.5 microg/ml, respectively, at 4 h 15, 1.2, 45 and 0.5 microg/ml, respectively, and at 24 h 0.2, 0.1, 1.8 and 0.1 microg/ml, respectively. All tumours displayed concentration dependent inhibition of colony formation after exposure to gemcitabine for 2 h. The IC(50)values of gemcitabine in six of the 10 metastases were > B 13%, Dukes' C 6%) and VIP (50%, 23%, 20%, 17%). For TH there was extensive loss of innervation around tumours of all stages (69%, 5%, 7%, 0%). SP immunoreactive peri-arteriolar nerves were similar in control tissue (39%) and tissue adjacent to Dukes' A tumours (40%) but diminished to 19% and 0% in tissue adjacent to Dukes' B and C tumours, respectively. In none of the tissues was CGRP immunoreactivity above 4%. The mean distance over which there was reduced NPY immunoreactivity from the tumour edge was 2.43 mm for Dukes' A/B tumours compared with 7.20 mm for Dukes' C tumours; for VIP immunoreactivity this distance was 5.22 mm for Dukes' A/B tumours and 5.52 mm for Dukes' C tumours. CONCLUSIONS: The progressive loss, both in terms of vascular nerve immunoreactivity and distance from the tumour edge with tumour grade, suggests that the tumour itself may influence neural integrity in perivascular plexuses, perhaps via the secretion of an inhibitory factor. PMID- 11034812 TI - Head and neck skin involvement by non-cutaneous head and neck cancers: free flap reconstruction. AB - INTRODUCTION: Patients with skin involvement from head and neck cancer have a poor prognosis, with a median survival time of 2 months. METHOD AND RESULTS: During a 9 year period, 31 patients with skin involvement above the clavicle by non-cutaneous malignant tumours of the head and neck were treated. In 19 males and 12 females with a mean age of 62 years, the parotid gland (32%) and the oral cavity (29%) were the commonest sites of primary disease and 77% of the cases were squamous cell carcinomas. Twenty-six had recurrent disease, 20 had received previous radiotherapy and all underwent surgical resection with free flap reconstruction, the commonest being the radial forearm (78%). Complete histological clearance was achieved in 53% of the cases, and adjuvant post operative irradiation was given to 60%. With this form of management, palliation was extended to a mean survival of 23 months. Six patients are currently alive and disease free at a mean follow up of 4.5 years. CONCLUSIONS: Patients with head and neck skin involvement by non-cutaneous head and neck malignancies have a very poor prognosis, but surgical resection combined with free microvascular flap reconstruction and planned post-operative radiotherapy, can offer good long-term palliation. PMID- 11034813 TI - EDUCATIONAL SECTION: risk analysis in surgical oncology-part I: concepts and tools. AB - All clinical procedures invoke risk. Many interventions in cancer management carry a particularly high element of risk, expressed through morbidity and premature death. Formal risk analysis is a discipline which is fundamental to engineering, to finance, to the airline industry and many other sectors of public life. Clinical risk analysis involves risk prediction, risk management and risk avoidance. Risk analysis is rarely invoked or taught in the clinical sciences, and management appraisals on individual patients almost never include a formal estimate of risk. Clinical decisions tend to be guided by qualitative judgements, and by the personality interactions of patients and clinicians. A formal evaluation of risk on a case by case and procedural basis might reduce morbidity and cost in surgical oncology practice. This article introduces the concepts, the spectrum and history of risk analysis and the tools for risk prediction. PMID- 11034814 TI - Novel chemotherapeutic agents in colorectal cancer. AB - Colorectal cancer remains the second most common cause of cancer death in Europe. A number of new agents, with varying mechanisms of action, have been developed and are being evaluated, both as single agents and in combination. Irinotecan, a topisomerase I inhibitor, and oxaliplatin, a new platinum compound, have proven efficacy in advanced colorectal cancer resistant to 5-fluorouracil. Capecitabine and other oral fluoro-pyrimidines are also emerging as potentially effective drugs. Raltitrexed and other inhibitors of thymidylate synthase (TS) are entering clinical practice although their role has yet to be determined. The new fluoro pyrimidines and TS inhibitors are important classes of drug which have been designed to take advantage of the knowledge of folate metabolism gained from basic clinical research. Their design features could perhaps reduce the toxicity seen with the first generation cytotoxic agents. This review will focus on these new chemotherapeutic agents in colorectal cancer with respect to their mechanisms of action, current clinical activity and their potential use both in the adjuvant and palliative settings. PMID- 11034815 TI - The platysma muscle in neck dissection. AB - Although the platysma muscle is usually preserved during neck dissection, removal of this muscle is generally considered inconsequential. The present case report shows that sacrifice of the cervical branch of the facial nerve innervating this muscle and removal of the platysma's upper portion impairs the caudal retraction and eversion of the ipsilateral half of the lower lip in grinning or laughing. PMID- 11034816 TI - Perianal extramammary Paget's disease. AB - Perianal extramammary Paget's disease is a rare disorder of unknown aetiology and is frequently associated with malignancy. Since Sir James Paget's original description, extramammary Paget's disease has been surrounded by controversy, speculation and much interest on the part of surgeons, pathologists and dermatologists. This case report draws attention to this rare condition, which should be considered in the differential diagnosis of perianal disorders. The physician must maintain a high index of suspicion, especially in cases with characteristic lesions unresponsive to conventional dermatological therapy. The importance of awareness of the condition is stressed and the diagnosis and treatment are commented upon. Treatment and prognosis depend on the presence of an underlying invasive carcinoma. An adequate initial evaluation and long-term follow up are necessary to identify recurrence and development of other malignancies. PMID- 11034817 TI - Quadruple cancer in a single patient: a report of four cases. AB - Multiple primary neoplasia was once considered a rare curiosity but is now a well recognized phenomenon. Only a few papers have been published in the English literature with regard to occurrence of four or more primary malignancies in a single patient. We report four cases of quadruple cancer; a review of the literature about this topic is discussed. PMID- 11034818 TI - REPORT: breast cancer conference report: ASCO (20-23 May 2000). PMID- 11034819 TI - Axillary recurrence of breast cancer following negative sentinel lymph node biopsy and axillary clearance. PMID- 11034821 TI - Response to letter by Dr van geel PMID- 11034820 TI - Comparison between low and high pressure suction drainage following axillary clearance. PMID- 11034822 TI - Peroperative hyperthermic intraperitoneal chemotherapy (HIPEC) for advanced gastric cancer. PMID- 11034823 TI - De bree, witkamp AJ, zoetmulder FAN. Peroperative hyperthermic intraperitoneal chemotherapy (HIPEC) for advanced gastric cancer PMID- 11034824 TI - Colorectal cancer follow-up. PMID- 11034825 TI - A multi-stage model of adenoma development. AB - The overwhelming proportion of colorectal carcinomas are believed to originate as adenomatous polyps (adenomas), and the identification and removal of adenomas is an important component of colorectal cancer prevention efforts. Mathematical modeling of adenomas can increase our understanding of the natural history and biology of adenomas and colorectal cancer and can help in the effort to devise optimal prevention and screening strategies. Here we adapt the multi-stage model of carcinogenesis to the problem of the development and growth of adenomas. We show that, using plausible values for the biological parameters, the model can fit various aspects of adenoma data including adenoma prevalence by age, the size distribution of adenomas, clustering of adenomas within individuals and the correlation between distal and proximal adenomas. Explaining the clustering of adenomas within individuals, as well as other findings, requires heterogeneity in risk in the population; we show how such heterogeneity can be related to the distribution of biological parameters in the population. The model can also be adapted to account for adenoma development in two major syndromes related to colorectal cancer, familial adenomatous polyposis and hereditary non-polyposis colorectal cancer. PMID- 11034826 TI - Kinetic proofreading scanning models for eukaryotic translational initiation: the cap and poly(A) tail dependency of translation. AB - Two simplified kinetic proofreading scanning (KPS) models were proposed to describe the 5' cap and 3' poly(A) tail dependency of eukaryotic translation initiation. In Model I, the initiation factor complex starts scanning and unwinding the secondary structure of the 5' untranslated region (UTR) from the 5' terminus of mRNA. In Model II, the initiation factor complex starts scanning from any binding site in the 5' UTR. In both models, following ATP hydrolysis, the initiation factor complex either dissociates from mRNA or continues to scan and unwind RNA secondary structure in the 5' UTR. This step repeats n times until the AUG codon is reached. These two models show very different cap and/or poly(A) tail dependency of translation initiation. The models predict that both cap and poly(A) tail dependencies of translation, and translatability of mRNAs are coupled with the structure of 5' UTR: the translation of mRNA with structured 5' UTR is strongly cap- and poly(A) tail-dependent; while translation of mRNA with unstructured 5' UTR is less cap- and poly(A) tail-dependent. We use these two models to explain: (1) the cap and poly(A) tail dependence of translation; (2) the effect of exogenous poly(A) on translation; (3) repression of host mRNA and translation of late adenovirus mRNA in the late phase of adenovirus infection; (4) repression of host mRNA and translation of Vaccinia virus mRNA in virus infected cell; (5) heat shock repression of translation of normal mRNA and stimulation of translation of hsp mRNA; and (6) the synergistic effect of cap and poly(A) tail on stimulating translation. The kinetic proofreading scanning models provide a coherent interpretation of those phenomena. PMID- 11034827 TI - Optimal sex ratios in structured populations. AB - In this paper, we develop a general method to determine evolutionary equilibrium sex ratios and to check evolutionary stability, continuous stability and invadability in exact genetic models with or without dominance. This method is then applied to three kinds of models for structured populations: the first one concerns Hamilton's LMC model, except that only a fraction beta of female offspring mate with male offspring born in the same colonies, while a fraction 1 beta mate with male offspring chosen at random within the whole population; in the second model, it is assumed that partial dispersal of inseminated females occurs after mating; in the third model, partial dispersal of male and female offspring occurs before mating. In the first model, the effect of population regulation is studied while, in the other models, two kinds of dispersal are considered: proportional and uniform. PMID- 11034828 TI - A statistical mechanical model for predicting B-DNA curvature and flexibility. AB - A statistical mechanical model taking into account the symmetric twisting, tilting, sliding fluctuations and asymmetric rolling fluctuations has been proposed to predict the macroscopic curvature and flexibility of B-DNA. Based on the statistical data of structural parameters of double helix in nucleic acid database and the related theoretical analysis, the equilibrium angular parameters (Omega, rho and tau) describing the orientation of successive base-pair planes, the translation parameters (D(y)) along the long axis of neighboring base-pair step and the corresponding force constants are arranged for ten dimers appropriately. Under the assumption of independent angular parameters, independent base-pair steps and a simple energy function, we can calculate the macroscopic curvature and the flexibility of DNA sequences through the transformation matrix and the Boltzmann ensemble average. The predictions on curvature and flexibility of DNA have been compared with the corresponding experimental data. The agreement is remarkably good. It is demonstrated that the lowering of the temperature does increase the DNA curvature. PMID- 11034829 TI - Designing lymphocyte functional structure for optimal signal detection: voila, T cells. AB - One basic task of immune systems is to detect signals from unknown "intruders" amidst a noisy background of harmless signals. To clarify the functional importance of many observed lymphocyte properties, I ask: What properties would a cell have if one designed it according to the theory of optimal detection, with minimal regard for biological constraints? Sparse and reasonable assumptions about the statistics of available signals prove sufficient for deriving many features of the optimal functional structure, in an incremental and modular design. The use of one common formalism guarantees that all parts of the design collaborate to solve the detection task. Detection performance is computed at several stages of the design. Comparison between design variants reveals e.g. the importance of controlling the signal integration time. This predicts that an appropriate control mechanism should exist. Comparing the design to reality, I find a striking similarity with many features of T cells. For example, the formalism dictates clonal specificity, serial receptor triggering, (grades of) anergy, negative and positive selection, co-stimulation, high-zone tolerance, and clonal production of cytokines. Serious mismatches should be found if T cells were hindered by mechanistic constraints or vestiges of their (co-)evolutionary history, but I have not found clear examples. By contrast, fundamental mismatches abound when comparing the design to immune systems of e.g. invertebrates. The wide-ranging differences seem to hinge on the (in)ability to generate a large diversity of receptors. PMID- 11034830 TI - Numerical methods and parameter estimation of a structured population model with discrete events in the life history. AB - We consider two numerical methods for the solution of a physiologically structured population (PSP) model with multiple life stages and discrete event reproduction. The model describes the dynamic behaviour of a predator-prey system consisting of rotifers predating on algae. The nitrate limited algal prey population is modelled unstructured and described by an ordinary differential equation (ODE). The formulation of the rotifer dynamics is based on a simple physiological model for their two life stages, the egg and the adult stage. An egg is produced when an energy buffer reaches a threshold value. The governing equations are coupled partial differential equations (PDE) with initial and boundary conditions. The population models together with the equation for the dynamics of the nutrient result in a chemostat model. Experimental data are used to estimate the model parameters. The results obtained with the explicit finite difference (FD) technique compare well with those of the Escalator Boxcar Train (EBT) method. This justifies the use of the fast FD method for the parameter estimation, a procedure which involves repeated solution of the model equations. PMID- 11034831 TI - Modelling T-cell-mediated suppression dependent on interactions in multicellular conjugates. AB - Tolerance to peripheral body antigens involves multiple mechanisms, namely T-cell mediated suppression of potentially autoimmune cells. Recent in vivo and in vitro evidence indicates that regulatory T cells suppress the response of effector T cells by a mechanism that requires the simultaneous conjugation of regulatory and effector T cells with the same antigen-presenting cell (APC). Despite this strong requirement, it is not yet clear what happens while both cells are conjugated. Several hypotheses are discussed in the literature. Suppression may result from simple competition of regulatory and effector cells for activation resources on the APC; regulatory T cells may deliver an inhibitory signal to effector T cells in the same conjugate; or effector T cells may acquire the regulatory phenotype during their interaction with regulatory T cells. The present article tries to further our understanding of T-cell-mediated suppression, and to narrow-down the number of candidate mechanisms. We propose the first general formalism describing the formation of multicellular conjugates of T cells and APCs. Using this formalism we derive three particular models, representing alternative mechanisms of T-cell-mediated suppression. For each model, we make phase plane and bifurcation analysis, and identify their pros and cons in terms of the relationship with the large body of experimental observations on T-cell-mediated suppression. We argue that accounting for the quantitative details of adoptive transfers of tolerance requires models with bistable regimes in which either regulatory cells or effectors cells dominate the steady state. From this analysis, we conclude that the most plausible mechanism of T-cell-mediated suppression requires that regulatory T cells actively inhibit the growth of effector T cells, and that the maintenance of the population of regulatory T cells is dependent on the effector T cells. The regulatory T cell population may depend on a growth factor produced by effector T cells and/or on a continuous differentiation of effector cells to the regulatory phenotype. PMID- 11034832 TI - DNA topology in heterochromatin (a hypothesis). AB - A hypothesis explaining the known heterochromatin features--a compact DNA packaging, transcriptional inactivity, propensity to aggregate (stickiness) and position effect variegation-is described. The hypothesis is based on the assumption that DNA molecules in heterochromatin are topologically open and contain single-strand breaks in the regions with identical or similar primary sequences. PMID- 11034833 TI - Design of large metabolic responses. Constraints and sensitivity analysis. AB - Metabolic control analysis (Kacser & Burns (1973). Symp. Soc. Exp. Biol.27, 65 104; Heinrich & Rapoport (1974). Eur. J. Biochem.42, 89-95) has been extensively used to describe the response of metabolic concentrations and fluxes to small (infinitesimal) changes in enzyme concentrations and effectors. Similarly, metabolic control design (Acerenza (1993). J. theor. Biol.165, 63-85) has been proposed to design small metabolic responses. These approaches have the limitation that they were not devised to deal with large (non-infinitesimal) responses. Here we develop a strategy to design large changes in the metabolic variables. The only assumption made is that, for all the parameter values under consideration, the system has a unique stable steady state. The procedure renders the kinetic parameters of the rate equations that when embedded in the metabolic network produce the pattern of large changes in the steady-state variables that we aim to design. Structural and kinetic constraints impose restrictions on the type of responses that could be designed. We show that these conditions can be transformed into the language of mean-sensitivity coefficients and, as a consequence, a sensitivity analysis of large metabolic responses can be performed after the system has been designed. The mean-sensitivity coefficients fulfil conservation and summation relationships that in the limit reduce to the well known theorems for infinitesimal changes. Finally, it is shown that the same procedure that was used to design metabolic responses and analyse their sensitivity properties can also be used to determine the values of kinetic parameters of the rate laws operating "in situ". PMID- 11034834 TI - A mathematical expectation model for bird navigation based on the clock-and compass strategy. AB - We present here a mathematical formula for the directional distribution of migratory birds if they use a vector navigation/clock-and-compass strategy to find their winter quarters. It is based on mathematical expectation theory and shows that a simple parabola can describe the expected geographical spread of clock-and-compass birds as a function of migratory distance. Predictions based on this model are then tested against all same autumn ringing recoveries of first season Pied Flycatchers, Ficedula hypoleuca, ringed in Scandinavia and European Robins, Erithacus rubecula, ringed in Sweden and Finland and recovered north of the Sahara Desert. We find that the predictions of our analytical model fit the ringing recovery distribution of freely migrating conspecifics extremely well. PMID- 11034835 TI - Energy expenditure of heavy to severe exercise and recovery. AB - A method of indirect calorimetry is proposed that attempts to better quantify the energy expenditure associated with heavy/severe exercise and the recovery from that exertion. To accomplish this objective, the energy expenditure associated with rapid anaerobic glycolysis is separated from that of mitochondrial respiration both during and after heavy/severe exercise. This model contrasts with those hypotheses that employ oxygen uptake as the sole measure of energy expenditure (e.g. the oxygen debt) or that utilizing a measure of anaerobic energy expenditure while ignoring the recovery energy expenditure. Anaerobic metabolism and its energy promoting effect on oxidative recovery must be independently acknowledged regardless of the eventual fate of lactate. PMID- 11034836 TI - A Modified Vibron Model for Anharmonic Vibrations: Application to Rovibrational Levels of Linear XYZ and XY(2) Molecules. AB - The vibron model for anharmonic vibrations of simple polyatomic molecules has been modified by explicit inclusion of the bending energy in the vibron hamiltonian, and by incorporating vibrationally and rotationally dependent terms into the main parameters of the model. These modifications keep the simple form of the basic vibron energy expressions unchanged but make them far more flexible and capable of much higher precision than those of the pure algebraic vibron model, even one with all squares and binary products of the Casimir operators included. Applications of the modified equations to the vibrational levels and rovibrational energies of the linear molecules HCN, CO(2), and some of their isotopomers show that this modified model yields precision of fitting which is better by one to two orders of magnitude than that provided by the strict algebraic model. This suggests that the modified vibron model can provide a simple but useful alternative to the much more elaborate global treatments of the rovibrational data. Copyright 2000 Academic Press. PMID- 11034837 TI - Fourier Transform Spectroscopy of the O(2) Herzberg Bands. III. Absorption Cross Sections of the Collision-Induced Bands and of the Herzberg Continuum. AB - Absorption spectra of molecular oxygen were measured in the laboratory under temperature and pressure conditions prevailing in the Earth's atmosphere. Spectra of pure O(2), O(2) + N(2), and O(2) + Ar were recorded in the 41 700 to 33 000 cm(-1) region (240-300 nm) at a maximal optical path difference of 0.45 cm using a Fourier transform spectrometer and a multiple reflection gas cell. The different components of the spectra, namely the discrete bands of the three Herzberg systems, the Herzberg continuum, and the collision-induced diffuse Wulf bands, were separated. The contribution of the Herzberg bands was first subtracted using the line parameters determined previously [A. Jenouvrier, M.-F. Merienne, B. Coquart, M. Carleer, S. Fally, A. C. Vandaele, C. Hermans, and R. Colin, J. Mol. Spectrosc. 198, 136-162 (1999)] from high-resolution data. Spectra recorded at various pressures then made it possible to determine by linear regression the intensity of the Wulf bands and the Herzberg continuum. The characteristics of the Wulf bands have been investigated in details: vibrational analysis, pressure effect, foreign gas effect, and a simulated spectrum are reported. The Herzberg continuum cross section is determined below the dissociation limit. A comparison with literature data is given. The new O(2) absorption cross sections and O(2)-O(2) collision-induced absorption cross sections are useful in connection with atmospheric measurements of ozone and other trace gases in the UV spectral region. Copyright 2000 Academic Press. PMID- 11034838 TI - High-Resolution Fourier Transform Emission Spectroscopy of the B(2)Sigma(+) (v = 0)-X(2)Sigma(+) (v = 0) Transition of the PN(+) Ion. AB - Ultraviolet emission spectrum of the B(2)Sigma(+) (v = 0)-X(2)Sigma(+) (v = 0) transition of the PN(+) ion has been observed at a resolution of 0.05 cm(-1) by a Fourier transform spectrometer. The rotational analysis led to a great improvement of rotational constants in the B(2)Sigma(+) (v = 0) and X(2)Sigma(+) (v = 0) states as well as to the first determination of centrifugal distortion constants for both levels and spin-rotation interaction constant in the X(2)Sigma(+) (v = 0) state. Rotational perturbation in the B(2)Sigma(+) (v = 0) state was observed. Copyright 2000 Academic Press. PMID- 11034839 TI - Generation, Microwave Spectrum, Barrier to Internal Rotation of Methyl Group, and ab Initio MO Calculation of syn-2-Nitrosopropene, syn CH(2)&dbond;C(CH(3))&bond;N&dbond;O. AB - syn-2-Nitrosopropene was generated, in the gas phase, by chemical reaction of 1 chloro-2-(hydroxyimino)propane with K(2)CO(3) and identified by microwave spectroscopy. The microwave spectrum of the reaction product was observed in the frequency range from 8.0 to 40.0 GHz. The rotational constants (MHz) were determined as A = 8744.09(6), B = 4846.07(2), and C = 3177.84(3) for CH(2)&dbond;C(CH(3))&bond;(14)NO (normal species) and A = 8664.36(5), B = 4822.15(3), and C = 3157.04(3) for CH(2)&dbond;C(CH(3))&bond;(15)NO ((15)N species) in the ground vibrational state. The values of the planar moment (P(cc) = (I(a) + I(b) - I(c))/2) obtained for the normal and (15)N species were 1.525(1) and 1.526(1) u A(2), respectively. This suggests that the nitrogen atom lies in or is close to the ab inertial plane of the molecule and shows also that only two hydrogen atoms are located symmetrically out of the symmetry plane. The reaction product was determined to be syn-2-nitrosopropene by comparing the observed and calculated rotational constants, kappa (Ray's asymmetry parameter) and r(s) coordinates of the nitrogen atom. The dipole moments (D) were determined to be u(a) = 2.43(5), u(b) = 1.12(7), and u(total) = 2.67(7). The barrier heights of the internal rotation owing to the methyl group of the normal species in the ground and first excited torsional states were determined to be 1750(50) and 1740(50) cal/mol (1 cal/mol = 4.184 J/mol), respectively. The (14)N nuclear quadrupole coupling constants (MHz) were determined to be chi(aa) = 0.25(21), chi(bb) = -7.11(40), and chi(cc) = 6.85(61). Two vibrational excited states were observed and the vibrational frequencies (cm(-1)) of the C-N and C-C torsional modes were determined to be 160(40) and 175(40), respectively. The lifetime of syn-2-nitrosopropene was found to be ca. 2 min in the waveguide cell. Copyright 2000 Academic Press. PMID- 11034840 TI - Absolute nu(2) Line Intensities of HOCl by Simultaneous Measurements in the Infrared with a Tunable Diode Laser and Far-Infrared Region Using a Fourier Transform Spectrometer. AB - We have measured absolute line intensities in the nu(2) fundamental band at 1238 cm(-1) of both isotopomers of hypochlorous acid, HOCl. To obtain the partial pressure of the species in the sample mixture, unavailable through direct measurement since HOCl exists only in equilibrium with H(2)O and Cl(2)O and may decay by secondary reactions, we relied on known absolute line intensities in the pure rotational far-infrared (FIR) spectrum determined from Stark effect measurements. We have thus recorded simultaneously the FIR pure rotation spectrum of HOCl using a Bruker IFS120HR interferometer and the spectrum of a few vibration-rotation lines in the infrared (IR) nu(2) band using a tunable diode laser spectrometer. The absolute intensities of these IR lines thus determined allowed us to "calibrate" the intensities of vibration-rotation lines in the whole nu(2) band, measured previously using Fourier transform spectroscopy. The treatment of the data took into account the blackbody emission contribution in the FIR and the evolution of the HOCl amount during the recording of the spectra. The latter was found to be almost constant over hours after conditioning of the cell. The square of the nu(2) band vibrational transition dipole moment was determined to be 0.013947(23) D(2) and 0.013870(51) D(2) for HO(35)Cl and HO(37)Cl, respectively, that is, 29 to 73% lower than previous measurements. A linear Herman-Wallis factor was also determined for both isotopomers. Finally, the line intensities were least-squares fitted using a model that takes into account a weak resonance between the (010) and (002) levels. Copyright 2000 Academic Press. PMID- 11034841 TI - LaseRitz: Far-Infrared Laser Line Assignment and Prediction by the Ritz Combination Principle, with Application to Methanol and Hydrazine. AB - A "LaseRitz" program is described for the systematic assignment and prediction of far-infrared laser (FIRL) transitions for a given molecule, such as methanol or hydrazine, which can be generated by optical pumping with known laser lines. The input data set for the program consists of identified molecular energy levels in ground and excited vibrational states, as compiled by our Ritz assignment program from the analysis of infrared (IR) bands and the FIR ground state spectrum. The program scans the data set for appropriate IR matches with an input list of laser pump lines for CO(2), N(2)O, etc., and generates an output table of all possible FIRL lines consistent with the molecular selection rules and lying within a prescribed wavenumber range. Initial tests on CH(3)OH have led to three likely new FIRL assignments; for N(2)H(4), assignments have been found for 14 new FIRL transition systems and verified for five others. Lists have been produced (deposited as supplementary data with the JMS archive reached at www.idealibrary.com or available on request from the authors) of CH(3)OH and N(2)H(4) FIRL lines potentially accessible by optical pumping with regular, sequence-band, hot-band, and isotopic CO(2) lines, plus N(2)O lines for N(2)H(4). The systematic approach with the LaseRitz program is more global and rigorous than earlier line-by-line studies, permitting calculation of all FIRL wavenumbers to spectroscopic accuracy. Copyright 2000 Academic Press. PMID- 11034842 TI - The Near-Infrared (3)Phi --> (1)(3)Delta Transition of LaF. AB - Rotational analyses of 32 subbands of the near-infrared (3)Phi --> (1)(3)Delta system of LaF have been carried out, involving the levels v = 0-7 of the two states. The three subsystems were first treated as separate singlet systems, directly at equilibrium. Perturbations were revealed in the lower state, ascribed to spin-uncoupling interactions between (1)(3)Delta(3)(v) and (1)(3)Delta(2)(v + 1) levels. A 21 x 21 matrix representation at equilibrium of the complex of interacting levels (v = 0-6) was then constructed, each diagonal v-block corresponding to a triplet model of the rovibrational (3)Delta Hamiltonian. The observed perturbations were completely reduced in the harmonic oscillator approximation with an experimental value of the interaction parameter B(Delta)(0,1) = 0.01322(2) cm(-1). The wavenumbers of some 4500 lines of the system were fitted with an rms error of about 0.005 cm(-1). (Line lists are available in electronic form via http://www-obs.univ lyon1.fr/~ABernard/index.html.) Copyright 2000 Academic Press. PMID- 11034843 TI - Millimeter-Wave Spectra and Global Torsion-Rotation Analysis for the CH(3)OD Isotopomer of Methanol. AB - New millimeter-wave and microwave measurements for CH(3)OD have been combined with previous literature data and with an extended body of Fourier transform far infrared observations in a full global analysis of the first two torsional states (v(t) = 0 and 1) of the ground vibrational state. The fitted CH(3)OD data set contained 564 microwave and millimeter-wave lines and 4664 far-infrared lines, representing the most recent available information in the quantum number ranges J /=65 years) patients, men and women, smokers and nonsmokers, and patients with or without diabetes or hypertension. The relative effect was smaller, but absolute risk reduction was similar in patients with hypertension compared with those without hypertension. Relative risk reduction was significant in predefined categories of baseline lipid concentrations. Tests for interaction were not significant between relative risk reduction and baseline total cholesterol (5% to 95% range 177 to 297 mg/dL, 4.6 to 7.7 mmol/L), HDL cholesterol (27 to 58 mg/dL, 0.7 to 1.5 mmol/L), and triglyceride (74 to 302 mg/dL, 0.8 to 3.4 mmol/L) concentrations, analyzed as continuous variables. However, for LDL cholesterol, the probability values for interaction were 0.068 for the prespecified primary end point and 0.019 for the expanded end point. Relative risk reduction was similar throughout most of the baseline LDL cholesterol range (125 to 212 mg/dL, 3.2 to 5.5 mmol/L) with the possible exception of the lowest quintile of CARE/LIPID (<125 mg/dL) (relative risk reduction 5%, 95% CI 19% to -12%). CONCLUSIONS: Pravastatin treatment is effective in reducing coronary heart disease events in patients with high or low risk factor status and across a wide range of pretreatment lipid concentrations. PMID- 11034936 TI - Platelet Pl(A2) allele and incidence of coronary heart disease: results from the Atherosclerosis Risk In Communities (ARIC) Study. AB - BACKGROUND: The major platelet integrin glycoprotein IIb-IIIa plays a primary role in platelet aggregation and acute thrombus formation at the site of vascular injury. A genetic polymorphism of glycoprotein IIb-IIIa (Pl(A)) has recently been proposed as a potential genetic factor linking to platelet hyperaggregability and increased risk of myocardial infarction. Despite numerous, mostly nonprospective studies, the role of this polymorphism as a clinically relevant, inherited risk factor for coronary heart disease (CHD) is still controversial. The purpose of this study was to determine whether Pl(A2) is a risk factor for incident CHD and whether it is correlated with increased platelet activation in a case-cohort study nested within a prospective epidemiologic investigation. METHODS AND RESULTS: Blood samples were collected and processed from the Atherosclerosis Risk in Communities Study cohort at the baseline examination (1987 to 1989). They were stored at -80 degrees C. Pl(A1/A2) genotype and plasma beta-thromboglobulin levels were determined in 439 incident CHD cases and a reference cohort sample of 544 (of whom 18 were also CHD cases). The prevalence of the Pl(A2) allele was not different in cases versus noncases. No significant correlation between CHD risk factors and the Pl(A2) allele was noted either. Platelet activation, as measured by plasma beta-thromboglobulin levels, was not enhanced in individuals with the Pl(A2) allele. CONCLUSIONS: This prospective study indicates that healthy individuals carrying the Pl(A2) allele do not have an increased risk of CHD. PMID- 11034937 TI - Low-dose transdermal testosterone therapy improves angina threshold in men with chronic stable angina: A randomized, double-blind, placebo-controlled study. AB - BACKGROUND: Experimental studies suggest that androgens induce coronary vasodilatation. We performed this pilot project to examine the clinical effects of long-term low-dose androgens in men with angina. METHODS AND RESULTS: Forty six men with stable angina completed a 2-week, single-blind placebo run-in, followed by double-blind randomization to 5 mg testosterone daily by transdermal patch or matching placebo for 12 weeks, in addition to their current medication. Time to 1-mm ST-segment depression on treadmill exercise testing and hormone levels were measured and quality of life was assessed by SF-36 at baseline and after 4 and 12 weeks of treatment. Active treatment resulted in a 2-fold increase in androgen levels and an increase in time to 1-mm ST-segment depression from (mean+/-SEM) 309+/-27 seconds at baseline to 343+/-26 seconds after 4 weeks and to 361+/-22 seconds after 12 weeks. This change was statistically significant compared with that seen in the placebo group (from 266+/-25 seconds at baseline to 284+/-23 seconds after 4 weeks and to 292+/-24 seconds after 12 weeks; P:=0.02 between the 2 groups by ANCOVA). The magnitude of the response was greater in those with lower baseline levels of bioavailable testosterone (r=-0. 455, P:<0.05). There were no significant changes in prostate specific antigen, hemoglobin, lipids, or coagulation profiles during the study. There were significant improvements in pain perception (P:=0.026) and role limitation resulting from physical problems (P:=0.024) in the testosterone-treated group. CONCLUSIONS: Low-dose supplemental testosterone treatment in men with chronic stable angina reduces exercise-induced myocardial ischemia. PMID- 11034938 TI - Modifications of cardiac autonomic profile associated with a shift schedule of work. AB - BACKGROUND: Shift work is associated with an increased rate of cardiovascular diseases and accidents. Discordance between circadian rhythms of stress-related biological variables and the work-sleep schedule might explain the reduced efficiency of work. It is not clear whether a shift schedule of work may induce similar discordance in the 24-hour oscillatory pattern of the cardiac autonomic control in respect to the work-sleep periods. METHODS AND RESULTS: Twenty-two healthy male blue-collar shift workers underwent 24-hour ECG recordings during each of the 3 different shifts (first, 6 AM to 2 PM; second, 2 to 10 PM; third, 10 PM to 6 AM). Spectral analysis of heart rate variability over 24 hours provided the normalized markers of cardiac sympathetic (LF(nu)) and vagal (HF(nu)) modulation of the sinoatrial node activity and of the sympathovagal balance (LF/HF). LF(nu) and LF/HF exhibited 24-hour oscillations with different times of maximum and minimum in accordance with the working and sleeping periods, respectively. Lower values of LF(nu) and LF/HF suggestive of a reduced cardiac sympathetic modulation were present when the job task was performed at night compared with the values observed when the work was performed during morning and evening. CONCLUSIONS: Continuous weekly changes of time of maximum and minimum in the cardiac sympathetic and vagal autonomic control may play a role in the excessive rate of cardiovascular diseases in shift workers. The reduced values of the indexes of cardiac sympathetic modulation during night work might be related to the presence of sleepiness or diminished alertness, which in turn could facilitate errors and accidents. PMID- 11034939 TI - Cytomegalovirus seropositivity and C-reactive protein have independent and combined predictive value for mortality in patients with angiographically demonstrated coronary artery disease. AB - BACKGROUND: The role of inflammation in coronary artery disease (CAD) is being increasingly recognized. Markers of inflammation (eg, C-reactive protein [CRP]) and infection (eg, seropositivity to Chlamydia pneumoniae, cytomegalovirus [CMV], and Helicobacter pylori) have been proposed as risk factors for CAD, but these associations require further evaluation. METHODS AND RESULTS: We prospectively tested whether CRP levels and IgG seropositivity to C pneumoniae, CMV, and H pylori are predictors of subsequent mortality in 985 consecutive patients with angiographically demonstrated CAD (stenosis >/=70%). Patients were followed for an average of 2.7 years (range 1.5 to 4.0 years). Patients averaged 65 years of age; 77% were men; and 110 (11.2%) died during follow-up. CRP levels were significantly elevated in nonsurvivors compared with survivors (mean CRP 3.1 mg/dL versus 1.5 mg/dL, P:=0.003). After controlling for all known baseline variables, the 2nd and 3rd tertiles of CRP compared with the 1st produced a Cox hazard ratio (HR) for mortality of 2.4 (P:=0.001). Of the 3 infectious markers tested, only seropositivity to CMV (HR=1.9, P:<0.05) was predictive of mortality. The majority of mortality risk associated with elevated CRP or CMV seropositivity occurred when both risk factors were present (P: for trend <0.0001). Other independent predictors of increased risk of mortality were age (HR=1.07 per year, P:<0.0001), left ventricular ejection fraction (HR=0.97 per percent, P:<0.0001), and diabetes mellitus (HR=1.7, P:=0.02). CONCLUSIONS: CMV seropositivity and elevated CRP, especially when in combination, are strong, independent predictors of mortality in patients with CAD. This suggests an interesting hypothesis that a chronic, smoldering infection (CMV) might have the capacity to accelerate the atherothrombotic process. PMID- 11034940 TI - Participation of tyrosine phosphorylation in cytoskeletal reorganization, alpha(IIb)beta(3) integrin receptor activation, and aspirin-insensitive mechanisms of thrombin-stimulated human platelets. AB - BACKGROUND: Fibrinogen binding to the active conformation of the alpha(IIb)beta(3) integrin receptor (glycoprotein IIb/IIIa) and cytoskeletal reorganization are important events in platelet function. Tyrosine phosphorylation of platelet proteins plays an essential role in platelet signal transduction pathways. We studied the participation of tyrosine kinases on these aspects of platelet reactivity and their importance in cyclooxygenase (COX)-1 independent mechanisms in thrombin-stimulated human platelets. METHODS AND RESULTS: Using washed platelets from normal donors and tyrphostin-A47 and aspirin as tyrosine kinase and COX-1 inhibitors, respectively, we found that tyrphostin A47 downregulated (1) the thrombin-activated conformational change of alpha(IIb)beta(3), (2) actin polymerization and cytoskeletal reorganization, and (3) the quantity of tyrosine-phospho-rylated proteins associated with the reorganized cytoskeleton. The latter are important components of multimolecular signaling complexes. Concomitantly, platelet aggregation and secretion were significantly reduced. Aspirin did not affect receptor activation or tyrosine phosphorylation but did decrease the initial (30-second) burst of actin polymerization. Importantly, aspirin significantly amplified the inhibitory effect of tyrphostin-A47 on all aspects of platelet reactivity that we evaluated. CONCLUSIONS: Tyrosine protein phosphorylation is a regulatory control system of the inside-out mechanism of alpha(IIb)beta(3) activation and cytoskeletal assembly in thrombin-stimulated human platelets. Inhibition of these aspects of platelet function with tyrphostin-A47 is amplified when platelets are treated with aspirin. Therefore, tyrosine phosphorylation is a major component of early signaling events and of COX-1-independent mechanisms of thrombin-induced platelet reactivity. The study results may indicate a novel target for therapeutic intervention. PMID- 11034941 TI - P-selectin expression on platelets determines size and stability of platelet aggregates. AB - BACKGROUND: P-selectin mediates rolling of platelets and leukocytes on activated endothelial cells. After platelet activation, P-selectin is translocated from intracellular granules to the external membrane, whereas fibrinogen aggregates platelets by bridging glycoprotein (GP) IIb/IIIa between adjacent platelets. METHODS AND RESULTS: In this study, we define a novel role for P-selectin in platelet aggregation. Expression of P-selectin on the platelet surface correlated strongly with the mean platelet aggregate size. Inhibition of P-selectin binding to its ligand by either monoclonal anti-P-selectin antibodies directed against the lectin domain or soluble human P-selectin reversed platelet aggregation even when added up to 5 minutes after activation; however, fibrinogen binding to platelets was not affected. This deaggregating effect significantly reduced the maximal size and number of platelet aggregates. When added 1 minute after platelet activation, anti-P-selectin antibody achieved 95% to 100% of the deaggregating effect of EDTA, whereas the anti-GP IIb/IIIa antibody abciximab had no effect. Monoclonal antibodies against known P-selectin ligands, such as P selectin GP ligand-1 (PSGL-1) or GP Ib, had no effect on platelet aggregation, suggesting a different ligand for P-selectin in platelet aggregate stabilization. In kinetic studies, P-selectin was maximally expressed 10 minutes after platelet activation, whereas maximal activation of GP IIb/IIIa occurred within the first 10 seconds, suggesting that P-selectin operates after fibrinogen binding to activated GP IIb/IIIa. CONCLUSIONS: These results indicate that P-selectin interaction with a ligand, different from PSGL-1 or GP Ib, stabilizes initial GP IIb/IIIa-fibrinogen interactions, allowing the formation of large stable platelet aggregates. PMID- 11034942 TI - Acute endothelin-A receptor antagonism prevents normal reduction of myocardial ischemia on repeated balloon inflations during angioplasty. AB - BACKGROUND: Myocardial ischemia and reperfusion are associated with increased production of endothelin (ET)-1. METHODS AND RESULTS: We examined the effects of BQ-123, a selective ET(A) receptor antagonist, in 80 patients. All patients were randomly allocated to an intracoronary infusion of saline or BQ-123 (6 micromol/L over 20 minutes). The reference group consisted of 20 patients undergoing coronary angiography. BQ-123 produced a 10% (P:<0.005) increase in distal coronary artery diameter. The main study group consisted of 30 patients undergoing coronary angioplasty. All patients underwent a minimum of 3 balloon inflations (BIs). Surface and intracoronary electrocardiographic ST-segment shift as well as pain score were recorded at the end of each BI. BQ-123 or saline was given by intracoronary infusion between the second and the third BI in random allocation. In the saline group, intracoronary ST-elevation decreased from 1.26+/ 0.55 mV during the first BI to 0.77+/-0.56 mV during the third BI (P:<0.05) and the surface ST elevation decreased from 0.20+/-0.15 to 0.10+/-0.07 mV (P:<0.05). In the BQ-123 group, the respective values were 1.22+/-0.48 mV and 1.13+/-0.62 mV (intracoronary) and 0.17+/-0.18 and 0.17+/-0.21 mV (surface) (both P:=NS). The decrease in pain score was significantly higher in the saline group (F:=5.97, P:=0.004). In 30 patients (collateral circulation group), the angioplasty protocol was repeated with the use of a pressure guide wire. BQ-123 produced a significant (F:=3.30, P:=0.04) decrease in coronary wedge pressure. CONCLUSIONS: Acute ET(A) receptor antagonism prevents the normal reduction of myocardial ischemia on repeated BIs during angioplasty. This may be explained by a "steal" effect through coronary collaterals. PMID- 11034943 TI - A matrix metalloproteinase induction/activation system exists in the human left ventricular myocardium and is upregulated in heart failure. AB - BACKGROUND: Matrix metalloproteinases (MMPs) contribute to matrix remodeling in disease states such as tumor metastases. Extracellular matrix metalloproteinase inducer (EMMPRIN) has been reported to increase MMP expression, and membrane-type MMP or MT1-MMP has been implicated to activate MMPs. The present study examined whether and to what degree EMMPRIN and MT1-MMP were expressed in human left ventricular (LV) myocardium as well as the association with MMP activity and expression in dilated cardiomyopathy (DCM). METHODS AND RESULTS: LV myocardial zymographic MMP activity increased by >2-fold with both nonischemic DCM (n=21) and ischemic DCM (n=16) compared with normal (n=13). LV myocardial abundance of MMP-9 was increased with both forms of DCM. MMP-2 and MMP-3 were increased with nonischemic DCM. MMP-1 levels were decreased with both forms of DCM. EMMPRIN increased by >250% and MT1-MMP increased by >1000% with both forms of DCM. CONCLUSIONS: Increased LV myocardial MMP activity and selective upregulation of MMPs with nonischemic and ischemic forms of DCM occurred. Moreover, a local MMP induction/activation system was identified in isolated normal human LV myocytes that was upregulated with DCM. The control of MMP activation and expression in the failing human LV myocardium represents a new and potentially significant therapeutic target for this disease process. PMID- 11034944 TI - Homozygous mutation in cardiac troponin T: implications for hypertrophic cardiomyopathy. AB - BACKGROUND: Mutations in the gene that encode cardiac troponin T (cTnT) account for approximately 15% of cases of familial hypertrophic cardiomyopathy (HCM). These mutations are associated with a particularly severe form of HCM characterized by a high incidence of sudden death and a poor overall prognosis, despite subclinical or mild left ventricular hypertrophy. METHODS AND RESULTS: We evaluated a family with HCM and multiple occurrences of sudden death in children. DNA samples were isolated from peripheral blood or paraffin-embedded tissue, and all protein-encoding exons of the cTnT gene were sequenced. A mutation was identified in exon 11 and is predicted to substitute a phenylalanine-for-serine mutation at residue 179 (Ser(179)Phe) in cTnT. Both parents and 3 of 4 surviving and clinically unaffected children were heterozygous for this mutation; another clinically unaffected child did not carry the mutation. Genetic analysis of DNA from a child who died suddenly at age 17 years demonstrated he was homozygous for this mutation. A review of his echocardiogram revealed profound left and right ventricular hypertrophy. CONCLUSIONS: An homozygous Ser(179)Phe mutation in cTnT causes a severe form of HCM characterized by striking morphological abnormalities and juvenile lethality. In contrast, the natural history of the heterozygous mutation is benign. These studies emphasize the relevance of genetic diagnosis in hypertrophic cardiomyopathy and provide a new perspective on the clinical consequences of troponin T mutations. PMID- 11034945 TI - Genome-wide linkage analysis of systolic and diastolic blood pressure: the Quebec Family Study. AB - BACKGROUND: Blood pressure (BP), an important risk factor for coronary heart disease, is a complex trait with multiple genetic etiologies. While some loci affecting BP variation are known (eg, angiotensinogen), there are likely to be novel signals that can be detected with a genome scan approach. METHODS AND RESULTS: A genome-wide scan was performed in 125 random and 81 obese families participating in the Quebec Family Study. A multipoint variance-components linkage analysis of 420 markers (353 microsatellites and 67 restriction fragment length polymorphisms) revealed several signals (P:<0.0023) for systolic BP on 1p (D1S551, ATP1A1), 2p (D2S1790, D2S2972), 5p (D5S1986), 7q (D7S530), 8q (CRH), and 19p (D19S247). Suggestive evidence (0.00230.10 ng/mL (hazard ratio 6.85, 95% CI 3. 04 to 15.45). Total homocysteine level greater than median was also associated with mortality (hazard ratio 2.44, 95% CI 1.10 to 5.40). These hazard ratios did not change substantially after adjustment for other risk factors. Significant predictors for CVD events were baseline diabetes, cerebrovascular disease, serum glucose, and triglycerides. After adjustment, only glucose and triglycerides remained significantly related to CVD events (hazard ratio with 95% CI 1.33 [1.12 to 1.57] and 1.14 [1.04 to 1.26], respectively, for a 1-mmol/L increase in concentration). CONCLUSIONS: We conclude that total homocysteine and particularly cardiac troponin T are important predictors of mortality in patients with end-stage renal disease, whereas other laboratory variables and baseline disease status have less prognostic value. PMID- 11034947 TI - Oxidized LDL upregulates angiotensin II type 1 receptor expression in cultured human coronary artery endothelial cells: the potential role of transcription factor NF-kappaB. AB - BACKGROUND: We demonstrated earlier that angiotensin II (Ang II), by AT(1) receptor activation, upregulates oxidized LDL (ox-LDL) endothelial receptor LOX-1 gene expression and uptake of ox-LDL in human coronary artery endothelial cells (HCAECs). In this study, we investigated the regulation of Ang II receptors (AT1R and AT2R) by ox-LDL and the role of the redox-sensitive transcription factor NF kappaB in this process. METHODS AND RESULTS: HCAECs were incubated with ox-LDL for 24 hours. Ox-LDL (10 to 40 microg protein/mL) upregulated AT1R but not AT2R, mRNA, or protein. Ox-LDL degraded IkappaBalpha in cytoplasm and activated transcription factor NF-kappaB (P65) in HCAEC nuclear extract. Treatment of cells with the antioxidant alpha-tocopherol (10 to 50 micromol/L) attenuated ox-LDL mediated degradation of IkappaBalpha and activation of NF-kappaB (P65) and inhibited the upregulation of AT1R mRNA and protein. The role of NF-kappaB signal transduction was further examined by use of an NF-kappaB inhibitor, caffeic acid phenethyl ester (CAPE). Pretreatment of cells with CAPE inhibited ox-LDL-mediated degradation of IkappaBalpha and NF-kappaB activation and inhibited ox-LDL-induced upregulation of AT1R expression. Incubation of cells with both ox-LDL and Ang II increased cell injury, measured as cell viability and LDH release, compared with either ox-LDL or Ang II alone. alpha-Tocopherol as well as the specific AT1R blocker CV11974 (candesartan) attenuated the cell-injurious effects of ox-LDL. CONCLUSIONS: These observations suggest an important role of ox-LDL-mediated AT1R upregulation in cell injury. In this process, NF-kappaB activation seems to play a critical role in signal transduction. These findings provide a basis for the use of antioxidants and AT1R blockers in designing therapy of atherosclerosis. PMID- 11034948 TI - Na(+)/H(+) exchange inhibitor cariporide attenuates cell injury predominantly during ischemia and not at onset of reperfusion in porcine hearts with low residual blood flow. AB - BACKGROUND: This study investigated whether myocardial protection by inhibition of Na(+)/H(+) exchange (NHE) occurs during ischemia and/or during reperfusion. METHODS AND RESULTS: The left anterior descending coronary artery was occluded in 32 pigs for 60 minutes and then reperfused for 24 hours. Infarct sizes (nitroblue tetrazolium [NBT] stain, histology) were determined at the end of the experiments. An extracorporeal bypass was used to achieve a constant residual blood flow of 3 mL/min in the myocardium at risk during ischemia. The NHE-1 inhibitor cariporide or distilled water was infused into the extracorporeal bypass system. In group 1, active treatment was administered from the onset of ischemia until 10 minutes of reperfusion (n=8). In group 2, active treatment was infused during the first 30 minutes of ischemia only (n=8). The group 3 animals (n=8) received intracoronary cariporide after 45 minutes of ischemia until 10 minutes of reperfusion. The control animals (group 4, n=7) were treated similarly to group 1 animals, with the cariporide solution being replaced by distilled water. Infarct sizes of group 1 (NBT stain, 41.5+/-20%; histology, 44. 6+/-12%) and group 2 (NBT stain, 33.5+/-14%; histology 34.9+/-15%) differed significantly (at least P:=0.012) from infarct sizes of group 3 (NBT stain, 71.6+/-15%; histology, 69.2+/-12%) and the control group (NBT stain, 76+/-9%; histology 72.4+/-12%). Cariporide treatment in group 1 and group 2 significantly improved functional recovery after 24 hours of reperfusion. CONCLUSIONS: Myocardial protection by cariporide is predominantly achieved by NHE inhibition during ischemia and not during early reperfusion. PMID- 11034949 TI - Single-beat estimation of end-systolic elastance using bilinearly approximated time-varying elastance curve. AB - BACKGROUND: Although left ventricular end-systolic elastance (E(es)) has often been used as an index of contractility, technical difficulties in measuring volume and in changing loading conditions have made its clinical application somewhat limited. By approximating the time-varying elastance curve by 2 linear functions (isovolumic contraction phase and ejection phase) and estimating the slope ratio of these, we developed a method to estimate E(es) on a single-beat basis from pressure values, systolic time intervals, and stroke volume. METHODS AND RESULTS: In 11 anesthetized dogs, we compared single-beat E(es) with that obtained with caval occlusion. Although the decrease (but not the increase) in contractility (5.3 to 11.4 mm Hg/mL) and the change in loading conditions (3.7 to 34.0 mm Hg/mL) over wide ranges significantly altered the slope ratio, the estimation of E(es) was reasonably accurate (y=0.97 x 0.46, r=0. 929, SEE=2.1 mm Hg/mL). CONCLUSIONS: E(es) can be estimated on a single-beat basis from easily obtainable variables by approximating the time-varying elastance curve by a bilinear function. PMID- 11034950 TI - Effects of dopamine beta-hydroxylase inhibition with nepicastat on the progression of left ventricular dysfunction and remodeling in dogs with chronic heart failure. AB - BACKGROUND: Inhibition of dopamine beta-hydroxylase (DBH) results in a decrease in norepinephrine synthesis. The present study was a randomized, blinded, placebo controlled investigation of the long-term effects of therapy with the DBH inhibitor nepicastat (NCT) on the progression of left ventricular (LV) dysfunction and remodeling in dogs with chronic heart failure (HF). METHODS AND RESULTS: Moderate HF (LV ejection fraction [LVEF] 30% to 40%) was produced in 30 dogs by intracoronary microembolization. Dogs were randomized to low-dose NCT (0.5 mg/kg twice daily, n=7) (L-NCT), high-dose NCT (2 mg/kg twice daily, n=7) (H NCT), L-NCT plus enalapril (10 mg twice daily, n=8) (L-NCT+ENA), or placebo (PL, n=8). Transmyocardial (coronary sinus-arterial) plasma norepinephrine (tNEPI), LVEF, end-systolic volume, and end-diastolic volume were measured before and 3 months after initiating therapy. tNEPI levels were higher in PL compared with NL (86+/-20 versus 13+/-14 pg/mL, P:<0.01). L-NCT alone and L-NCT+ENA reduced tNEPI toward normal (28+/-4 and 39+/-17 pg/mL respectively), whereas HD-NCT reduced tNEPI to below normal levels (3+/-10 pg/mL). In PL dogs, LVEF decreased but was unchanged with L-NCT and increased with L-NCT+ENA. L-NCT and L-NCT+ENA prevented progressive LV remodeling, as evidenced by lack of ongoing increase in end diastolic volume and end-systolic volume, whereas H-NCT did not CONCLUSIONS: In dogs with HF, therapy with L-NCT prevented progressive LV dysfunction and remodeling. The addition of ENA to L-NCT afforded a greater increase in LV systolic function. NCT at doses that normalize tNEPI may be useful in the treatment of chronic HF. PMID- 11034951 TI - Calcineurin inhibitor attenuates the development and induces the regression of cardiac hypertrophy in rats with salt-sensitive hypertension. AB - BACKGROUND: It remains unclear how hemodynamic overload induces cardiac hypertrophy. Recently, activation of calcium-dependent phosphatase, calcineurin, has been elucidated to induce cardiac hypertrophy. In the present study, we examined the role of calcineurin in load-induced cardiac hypertrophy by using Dahl salt-sensitive (DS) rats, which develop both pressure and volume overload when fed a high salt diet. METHODS AND RESULTS: In the DS rat heart, the activity of calcineurin was increased and cardiac hypertrophy was induced by high salt diet. Treatment of DS rats with the calcineurin inhibitor FK506 (0.1 or 0.01 mg/kg twice daily) from the age of 6 weeks to 12 weeks inhibited the activation of calcineurin in the heart in a dose-dependent manner and attenuated the development of load-induced cardiac hypertrophy and fibrosis without change of hemodynamic parameters. Additionally, treatment with 0.1 mg/kg twice daily but not with 0.01 mg/kg twice daily of FK506 from the age of 12 weeks to 16 weeks induced regression of cardiac hypertrophy in DS rats. Load-induced reprogramming of gene expression was also suppressed by the FK506 treatment. CONCLUSIONS: These results suggest that calcineurin is involved in the development of cardiac hypertrophy in rats with salt-sensitive hypertension and that inhibition of calcineurin could induce regression of cardiac hypertrophy. PMID- 11034952 TI - Gene transfer of human prostacyclin synthase ameliorates monocrotaline-induced pulmonary hypertension in rats. AB - BACKGROUND: Prostacyclin is a potent vasodilator that also inhibits platelet adhesion and cell growth. We investigated whether in vivo gene transfer of human prostacyclin synthase (PGIS) ameliorates monocrotaline (MCT)-induced pulmonary hypertension in rats. METHODS AND RESULTS: The cDNA encoding PGIS was intratracheally transfected into the lungs of rats by the hemagglutinating virus of Japan-liposome method. Rats transfected with control vector lacking the PGIS gene served as controls. Three weeks after MCT injection, mean pulmonary arterial pressure and total pulmonary resistance had increased significantly; the increases were significantly attenuated in PGIS gene-transfected rats compared with controls [mean pulmonary arterial pressure, 31+/-1 versus 35+/-1 mm Hg ( 12%); total pulmonary resistance, 0.087+/-0.01 versus 0.113+/-0.01 mm Hg x mL x min(-1) x kg(-1) (-23%), both P:<0.05]. Systemic arterial pressure and heart rate were unaffected. Histologically, PGIS gene transfer inhibited the increase in medial wall thickness of peripheral pulmonary arteries that resulted from MCT injection. PGIS immunoreactivity was intense predominantly in the bronchial epithelium and alveolar cells. Lung tissue levels of 6-keto-PGF(1alpha), a stable metabolite of prostacyclin, were significantly increased for >/=1 week after transfer of PGIS gene. The Kaplan-Meier survival curves demonstrated that repeated transfer of PGIS gene every 2 weeks increased survival rate in MCT rats (log-rank test, P:<0.01). CONCLUSIONS: Intratracheal transfer of the human PGIS gene augmented pulmonary prostacyclin synthesis, ameliorated MCT-induced pulmonary hypertension, and thereby improved survival in MCT rats. PMID- 11034953 TI - Pulmonary capillary endothelium-bound angiotensin-converting enzyme activity in acute lung injury. AB - BACKGROUND: Pulmonary capillary endothelium-bound (PCEB) angiotensin-converting ectoenzyme (ACE) activity alteration is an early, sensitive, and quantifiable lung injury index in animal models. We hypothesized that (1) PCEB-ACE alterations can be found in patients with acute lung injury (ALI) and (2) PCEB-ACE activity correlates with the severity of lung injury and may be used as a quantifiable marker of the underlying pulmonary capillary endothelial dysfunction. METHODS AND RESULTS: Applying indicator-dilution techniques, we measured single-pass transpulmonary hydrolysis of the synthetic ACE substrate (3)H-benzoyl-Phe-Ala-Pro (BPAP) in 33 mechanically ventilated, critically ill patients with a lung injury score (LIS) ranging from 0 (no lung injury) to 3.7 (severe lung injury) and calculated the kinetic parameter A(max)/K(m). Both parameters decreased early during the ALI continuum and were inversely related to APACHE II score and LIS. Hydrolysis decreased with increasing cardiac output (CO), whereas 2 different patterns were observed between CO and A(max)/K(m). CONCLUSIONS: PCEB-ACE activity decreases early during ALI, correlates with the clinical severity of both the lung injury and the underlying disease, and may be used as a quantifiable marker of underlying pulmonary capillary endothelial dysfunction. PMID- 11034954 TI - Dual SPECT imaging of Loffler's endomyocarditis in the acute phase. PMID- 11034955 TI - Positive remodeling, regression of in-stent neointimal hyperplasia, and late stent malapposition in the absence of brachytherapy. PMID- 11034956 TI - US Department of Agriculture and animal rights group settle suit over mice, rats, and birds; Congress may intervene. PMID- 11034957 TI - Lifestyle risk factors for cancer: the relationship with psychosocial work environment. AB - BACKGROUND: Psychosocial work characteristics (job demands, control, support, job strain and iso-strain [high job strain combined with social isolation at work]) may be linked to cancer risk, by affecting cancer-related lifestyles like smoking, high alcohol consumption, low intake of fruits and vegetables and lack of physical activity. METHODS: Cross-sectional data obtained from 3309 respondents participating in an ongoing prospective cohort study in the Netherlands on psychosocial factors and cancer risk were used to study the association between psychosocial job characteristics and lifestyle. Information on job characteristics and risk factor prevalence was collected from 20-65-year old men and women, through self-administered questionnaires. Multiple logistic and linear regression analyses were undertaken by gender, with adjustment for age and education. RESULTS: No differences in the prevalence of lifestyle risk factors for cancer were found amongst the psychosocial work characteristics studied. Moreover, little evidence was found for a relation between job (or iso-) strain and cancer-related lifestyles in multivariate analyses. CONCLUSIONS: The present study found no support for the hypothesis that job strain or iso-strain are associated with a cancer-related lifestyle. Further research on the role of other psychosocial factors-like personality or social support outside work-in mediating associations between job characteristics and lifestyle, is warranted. PMID- 11034958 TI - Characteristics of respondents and non-respondents from a case-control study of breast cancer in younger women. AB - BACKGROUND: This study assessed the nature of potential biases by comparing respondents with non-respondents from a case-control study of breast cancer in younger women. METHODS: The case-control study was conducted in three regions in the US: Atlanta GA, Seattle/Puget Sound WA, and central New Jersey. An abbreviated interview or mailed questionnaire was completed by willing non respondents, most of whom had refused participation in the main study. RESULTS: Respondents and non-respondents appeared similar with respect to age, race, relative weight, smoking, family history of breast cancer, number of births, age at first birth, and several dietary items. Compared to non-respondents, case and control respondents were of shorter stature, and reported less frequent consumption of doughnuts/pastries. Respondent cases, compared with non-respondent cases, were more highly educated and more likely to have consumed alcohol regularly; similar but not statistically significant tendencies were observed for controls. Respondent cases experienced menarche earlier than non-respondents. Respondent controls were more likely to have used oral contraceptives than non respondents; a similar but not statistically significant tendency was observed in cases. Comparisons of crude and simulated relative risks using available non respondents' data generally showed a low impact of non-response on relative risks in this study. CONCLUSIONS: Our results suggest that non-response would not greatly affect relative risk estimates in this study, except possibly regarding height. However, we were limited by the numbers of informative non-respondents and the amount of data collected. Collecting similar information in future studies would be useful, especially since varying methods used to encourage participation may lead to differences in respondents' characteristics. PMID- 11034959 TI - Influence of menstrual and reproductive factors on ovarian cancer risk in women with and without family history of breast or ovarian cancer. AB - BACKGROUND: As women with a family history of ovarian and/or breast cancer possibly inherit genetic changes that alter their risk of ovarian cancer, other established risk factors for ovarian cancer may influence the risk differently in women with and without a family history of the disease. METHODS: Case-control study conducted between 1983 and 1991 in Northern Italy. Cases were 971 women, under 75 years, with incident, histologically confirmed epithelial ovarian cancer, and controls were 2758 women, under 75 years, admitted to hospitals for non-malignant, non-hormone-related conditions, who had not undergone bilateral oophorectomy. Of these, 93 cases and 139 controls had a family history of ovarian and/or breast cancer. RESULTS: The risk of ovarian cancer increased with irregular menstrual cycles, late age at menopause, natural menopause, nulliparity, never use of oral contraceptives and use of hormone replacement therapy. We computed an 'adult life risk score' (ALRS) considering the combined effect of these factors. Compared to women without a family history and a low ALRS, the OR was 1.7 for women without family history and high ALRS, 1.4 for women with a family history and low ALRS, and 3.5 for women with a family history and high ALRS. CONCLUSIONS: Intervention on selected hormonal risk factors for ovarian cancer might be important for women with a family history of the disease. PMID- 11034960 TI - A study on effectiveness of screening mammograms. AB - BACKGROUND: So far, no randomized controlled trials with a mean mammographic screening interval of > or = 2 years has demonstrated statistically significant mortality reduction for women younger than age 50. The issue of screening frequency is vital in detection of primary breast cancer. METHODS: The study group consisted of cancers diagnosed in women who participated in a serial screening programme with a mean screening interval of 2 years. To study the effectiveness of the screening, a comparison is made between the distribution of age at which the tumour could be detected when biennial mammographic screening is the only detection method, and the distribution of age at which the tumour would be detected by either biennial mammographic screening or the development of symptoms. Some recently developed statistic methods, such as bootstrap, the maximum likelihood distribution estimator for doubly censored data and the EM algorithm, are used in estimation of these distributions. RESULTS: The hypothesis tests and confidence intervals show that the difference between the two distributions was statistically significant for women younger than 50 and 50-70 years old, but not for women over 70 years. CONCLUSIONS: The statistical analysis indicates that for women younger than 50, and 50-70 years of age, a screening mammogram every other year is not frequent enough to detect primary breast cancer, but for women over 70 years, it might be sufficient. PMID- 11034961 TI - Selective screening for cervical neoplasia: an approach for resource-poor settings. AB - BACKGROUND: Cervical malignancies are the leading cause of cancer-related morbidity and mortality among women in developing countries. Although early detection programmes using cytological methods, followed by aggressive treatment of precursor lesions are accepted as the main disease control strategy, fiscal limitations make this strategy unfeasible in many countries. METHODS: To screen selectively, we developed two risk scores using data from a population-based case control study in Jamaica with 202 cases and 363 controls. Independent risk factors for cervical neoplasia were determined using logistic regression. An unweighted risk score for each subject was developed by a simple count of risk factors present and a weighted risk score was calculated by summing regression coefficients for each risk factor. RESULTS: Four patient characteristics were independently predictive of cervical neoplasia, older age (OR = 3.4, 95% CI : 1.8 6.7), > or = 4 pregnancies (OR = 5.6, 95% CI : 1.2-18.7), poverty (OR = 2.1, 95% CI : 1.3-3.3) and cigarette smoking (OR = 1.9, 95% CI : 1.2-3.2). Using cut-off points of > or = 20 for the weighted scores and > 3 for unweighted scores, the sensitivity and specificity were 65% and 69% for the unweighted score and 75% and 61%, respectively, for the weighted score. Areas under the receiver operating characteristic (ROC) curves for the weighted versus the unweighted scores were similar, suggesting similar overall accuracy. CONCLUSION: Selective screening using risk assessment strategies is potentially useful, particularly in resource poor settings. However, whether weighting factors is essential is dependent on prevalence of factors in a given setting. Although this approach needs validation in other populations, women at highest risk for cervical neoplasia can be identified using demographic factors available during a regular clinic visit. PMID- 11034962 TI - Cervical cancer mortality in Australia: contrasting risk by Aboriginality, age and rurality. AB - BACKGROUND: The poor health status of Australia's indigenous population is reflected in relatively high mortality rates from almost all causes, including preventable causes such as cervical cancer, where the rate is six to eight times that of non-Aboriginal women. However, there is little information on the geographical distribution of risk, an important issue for service deployment. This study examined the risk of death from cervical cancer in relation to Indigenous status, age and rurality. METHODS: Data from death registers from Australian states and territories who have identified Aboriginal people were examined for 1986-1997 to obtain a list of all deaths where the primary cause was cancer of the cervix. The data categorized females by 5-year age group, by metropolitan, rural or remote category and by Indigenous status. Mean age at death and standardized mortality ratios for deaths from cervical cancer were calculated for Aboriginal compared with non-Aboriginal women in metropolitan, rural and remote areas. RESULTS: The risk of death from cervical cancer for Aboriginal women compared with non-Aboriginal women increased by 4.3-fold for metropolitan areas, 9.7-fold for rural areas and 18.3-fold for remote areas. CONCLUSIONS: Aboriginal women in rural and remote areas of Australia are at significantly higher risk of death from cancer of the cervix than either Aboriginal women in metropolitan areas or non-Aboriginal women in any area. This result raises questions about access to services for prevention and early diagnosis and other factors that might impact on the incidence and natural history of the disease. PMID- 11034963 TI - Co-factors related to the causal relationship between human papillomavirus and invasive cervical cancer in Honduras. AB - BACKGROUND: A case-control study was conducted in Honduras to identify co-factors in the carcinogenic pathway by which human papillomavirus (HPV) causes invasive cervical cancer. METHODS: Ninety-nine cases aged 23-65 (median 47) years participated. Two controls were matched to each case by age and clinic where they first presented for cytological screening; controls had no cervical abnormalities. Information on risk factors was obtained by personal interviews in the clinics regarding sociodemographic, reproductive and behavioral characteristics. Human papillomavirus was detected in cervical scrapes by general primer-mediated polymerase chain reaction (PCR) followed by sequence analysis to identify the different types present. RESULTS: All cases had squamous cell tumours and most were FIGO (International Federation of Gynecologists and Obstetricians) class II or higher; HPV was strongly associated with cervical cancer (odds ratio [OR] = 7.66, 95% CI : 3.88-15.1). Among HPV-positive women, dose-response relationships were observed for education, age at first intercourse and exposure to wood smoke that persisted after adjustment for previous screening. Among HPV-negative women, the number of sexual partners and parity were associated with cervical cancer. The protective effect of previous cytological screening operated independently of HPV. CONCLUSIONS: Our findings speak for the powerful role that both primary and secondary education plays in fostering a lifestyle that reduces the risk of invasive cervical cancer. The data suggest that important elements of such a lifestyle include later age at first sexual intercourse, a limited number of pregnancies, greater likelihood of undergoing cytological screening and reduced exposure to carcinogens in the household environment. PMID- 11034964 TI - Clues to the aetiological heterogeneity of testicular seminomas and non seminomas: time trends and age-period-cohort effects. AB - BACKGROUND: Most previous epidemiological studies have treated testicular cancer as a single entity. However, some investigators suggest that testicular seminomas and non-seminomas may have different risk profiles. We examine the time trends in incidence of the two main histological types separately. METHODS: From 1970 through 1995, 7296 cases of testicular cancer were registered in the Canadian provinces of Ontario, Saskatchewan and British Columbia. In addition to analyses of the secular trends by age group and birth cohort, an age-period-cohort (APC) model with standard Poisson assumptions was fitted to the data to assess the time effects. RESULTS: The age-adjusted incidence rate for seminomas increased by 53%, from 1.5 per 100 000 males in 1970-1971 to 2.3 per 100 000 males in 1994-1995. Non-seminomas increased by 91%, from 1.1 to 2.1 per 100 000 males over the same period. Non-seminomas were more frequent at young ages whereas seminomas dominated in older ages. In contrast to seminomas, non-seminomas occurred predominantly among adolescent men (15-19 years), with a fourfold increase between 1970-1971 and 1994-1995. Age-period-cohort modelling showed that the increase in the risk of both seminomas and non-seminomas followed a birth cohort pattern, but with differences in birth cohorts in addition to significantly distinct age patterns. CONCLUSIONS: Our findings support the hypothesis postulating aetiological heterogeneity in the development of seminomas and non seminomas. We suggest that epidemiological studies of testicular cancer treat seminomas and non-seminomas separately. PMID- 11034965 TI - Ecological study of the association between soy product intake and mortality from cancer and heart disease in Japan. AB - BACKGROUND: The anticarcinogenic and antiatherogenic properties of soy have been demonstrated in experimental studies. To evaluate the relationship between soy product intake and mortality from several types of cancer and heart disease, an ecological analysis was performed in 47 prefectures in Japan. METHODS: Age standardized mortality rates for heart disease and stomach, colorectal, lung, breast and prostate cancer were obtained from the National Vital Statistics, 1995. Information on major nutrient and soy product intake was obtained from the National Nutritional Survey Report 1980-1985. In this survey, dietary habits were surveyed annually by 3-day diet record in about 6000 randomly selected households. RESULTS: Soy protein intake was significantly correlated with stomach cancer mortality rate in men after controlling for total energy, alcohol and salt intake, and the mean age and proportion of current smokers in the prefecture (r = -0.31, P = 0.04). Soy product intake estimated as total amount as well as isoflavone and soy protein intake were significantly positively correlated with colorectal cancer mortality rates in both sexes (for total amount, r = 0.32, P = 0.03 in men and r = 0.44, P = 0.001 in women) after controlling for covariates. The inverse correlation between soy product intake (as total amount or soy protein) and heart disease mortality rate was statistically significant in women after controlling for covariates (r = 0.32, P = 0.04 and r = -0.31, P = 0.045, respectively). CONCLUSION: The present study provides modest support for the preventive role of soy against stomach cancer and heart disease death. PMID- 11034966 TI - Comparison of fatal coronary heart disease occurrence based on population surveys in Japan and the USA. AB - BACKGROUND: Although vital statistics have indicated large Japanese-American differences in mortality rates for coronary heart disease (CHD), the magnitude of difference has not been documented well using comparable validation of cause of death. METHODS: Population-based fatal CHD data were compared between the Oita Cardiac Death Survey, Japan and the Atherosclerosis Risk in Communities (ARIC) Study, USA. Both studies (population: Oita City 198 093; the ARIC comunities 286 820) identified possible fatal CHD events (International Classification of Diseases, Ninth Revision [ICD-9]: 410-414, 250, 401-402, 427-429, 440, and 798 799) among residents aged 35-74 years during 1992-1993. Comparable criteria for classifying cause of death were applied. Sex-specific, age-adjusted mortality rates of CHD were calculated by place of death. RESULTS: In all, 330 deaths in Oita and 1398 in the ARIC communities had eligible ICD-9 death certificate codes; CHD codes (ICD-9 410-414) comprised 30.6% of investigated deaths in Oita and 58.6% in ARIC. For men, the non-validated rate ratio for CHD deaths (ARIC:Oita City) was 5.9 (95% CI : 4.2-8.5), which fell to 4.7 (95% CI : 3.5-6.4) with validation and inclusion of sudden deaths within one hour of onset as fatal CHD. For women, the overall non-validated rate ratio was 4.6 (95% CI : 2.8-7.6), which fell to 3.9 (95% CI : 2.4-6.3) with validation and but there was little further change when the sudden deaths were added. CONCLUSIONS: Our results suggest that differences in fatal CHD rates between Japanese and Americans were not as large as suggested by vital statistics when events were validated and sudden deaths were included. PMID- 11034967 TI - Smoking, body mass index, socioeconomic status and the menopausal transition in a British national cohort. AB - BACKGROUND: This study investigates whether cigarette smoking, body mass index (BMI) and socioeconomic status are independently associated with age at menopausal transition. METHODS: Menopausal status and risk factor information were collected prospectively from 1572 British women followed up since their birth in 1946, so far until 50 years. Cox's regression models were used to investigate the relationships of interest. RESULTS: Cigarette smokers started the perimenopause and reached the menopause earlier than ex-smokers and non-smokers. The relative risk for smokers compared with non-smokers was 1.31 (95% CI : 1.09 1.56) for perimenopause and 1.63 (95% CI : 1.17-2.27) for menopause. Body mass index was associated with the age at inception of the perimenopause only among smokers and ex-smokers, with underweight women having the earliest perimenopause. No association was observed between BMI and age at menopause. Smokers and underweight women were more likely than others to start hormone replacement therapy (HRT) before becoming postmenopausal. There was no effect of education or social class on age at inception of the perimenopause or age at menopause. Single women had an earlier perimenopause but the effect was confounded by parity. CONCLUSIONS: Smoking was independently related to an earlier menopausal transition, although the effect on inception of the perimenopause was particularly observed among underweight women. There was no independent effect of socioeconomic status. The popularity of HRT use in this cohort may have had an impact on the findings. PMID- 11034968 TI - Standardized mortality ratio and life expectancy: a comparative study of Chinese mortality. AB - BACKGROUND: Various models have been proposed for rapid conversion of the standardized mortality ratio (SMR) to life expectancy using data from developed countries. METHODS: We compared two methods for converting the SMR to life expectancy using mortality data from the largest developing country, China. RESULTS: The first model, using the Gompertz function, does not provide a good fit to the life expectancy and SMR of China. The regression lines derived from the second, a log-linear model using parameters estimated from the US white population are not a good fit to Chinese males and older females. However, if the parameters in the log-linear model are estimated using Chinese mortality data, the resultant regression lines fit the data reasonably well. CONCLUSION: The relationship between life expectancy and SMR based on mortality data from developed countries may not be valid for developing countries. Based on our empirical study, separate estimates of the coefficients of the model are required for developing countries. PMID- 11034969 TI - Time to pregnancy as a correlate of fecundity: differential persistence in trying to become pregnant as a source of bias. AB - BACKGROUND: Subfecundity is a frequent and often serious problem and it is important to identify its preventable determinants and to monitor fecundity over time. Since follow-up studies are difficult and expensive to conduct, time to pregnancy (TTP) in pregnant women is often used as a surrogate measure of fecundity. TTP data can be retrieved at low costs and they need no valid population registry as a source for sampling. While TTP may serve as a valid surrogate measure in many situations, its validity rests upon a number of assumptions. We have analysed one of these overlooked assumptions, the importance of persistence in trying to become pregnant. METHODS: By means of computer simulations we estimated bias caused by differences in persistence in pregnancy attempts. We investigated whether the assumptions made in the simulation were realistic by using empirical data from a European study. RESULTS: The mean waiting time to pregnancy and other estimates of subfecundity (or infertility) strongly depend upon the persistence of couples in pursuing a pregnancy. We show that even moderate changes in the planning behaviour considerably modify the waiting time distribution. Empirical data confirm that persistence in trying to become pregnant is age-related. CONCLUSIONS: Persistence in pregnancy attempts affects outcome measures of subfecundity in studies based upon TTP in pregnant women. It is likely that the length of time during which couples keep trying to become pregnant is influenced by a number of factors which would probably change over time or be different between populations to be compared. PMID- 11034970 TI - Effect of outdoor and indoor nitrogen dioxide on respiratory symptoms in schoolchildren. AB - BACKGROUND: Nitrogen dioxide (NO(2)), an oxidant gas that contaminates both outdoor and indoor air, is considered to be a potential risk factor for asthma. We investigated concurrently the effects of outdoor and indoor NO(2) on the prevalence and incidence of respiratory symptoms among children. METHODS: A cohort study was carried out over 3 years on 842 schoolchildren living in seven different communities in Japan. Indoor NO(2) concentrations over 24 hours were measured in both winter and summer in the homes of the subjects, and a 3-year average of the outdoor NO(2) concentration was determined for each community. Respiratory symptoms were evaluated every year from responses to questionnaires. RESULTS: The prevalence of bronchitis, wheeze, and asthma significantly increased with increases of indoor NO(2) concentrations among girls, but not among boys. In neither boys nor girls were there significant differences in the prevalence of respiratory symptoms among urban, suburban, and rural districts. The incidence of asthma increased among children living in areas with high concentrations of outdoor NO(2). Multiple logistic regression analysis showed that a 10 parts per billion (ppb) increase of outdoor NO(2) concentration was associated with an increased incidence of wheeze and asthma (odds ratios [OR] = 1.76, 95% CI : 1.04 3.23 and OR = 2.10, 95% CI : 1.10-4.75, respectively), but that no such associations were found with indoor NO(2) concentration (OR = 0.73, 95% CI : 0.45 1.14 and OR = 0.87, 95% CI : 0.51-1.43, respectively). CONCLUSIONS: These findings suggest that outdoor NO(2) air pollution may be particularly important for the development of wheeze and asthma among children. Indoor NO(2) concentrations were associated with the prevalence of respiratory symptoms only among girls. Girls may be more susceptible to indoor air pollution than boys. PMID- 11034971 TI - Estimating the burden of disease in one Swiss canton: what do disability adjusted life years (DALY) tell us? AB - BACKGROUND: Examining life expectancy and general mortality rates, the health of the population of Geneva can be described as one of the best in the world. However, in some areas Geneva fares worse than the rest of Switzerland or Europe. To re-appraise the current health priorities of the Genevan population, we analysed the relative importance of specific diseases and injuries calculating DALYs. METHODS: We followed the procedures developed for the Global Burden of Disease (GBD) study to ensure comparability. Some adaptations were made for mortality coding. Disability was estimated based on data for countries classified as Established Market Economies (EME) in the GBD study. RESULTS: Non-communicable diseases accounted for 79% of the disability adjusted life years (DALY), injuries represented 12%, and communicable diseases and other disorders 9%. Ischaemic heart disease was the largest single contributor to DALY, followed by unipolar major depression. Neuropsychiatric disorders and mental health accounted for more than 23% of DALY. CONCLUSIONS: Some of the most important problems identified depression, osteoarthritis and alcohol abuse-would have been overlooked in an analysis based solely on mortality data. The most striking finding is the importance of mental health problems. The main limitation is the lack of morbidity data for Geneva. PMID- 11034972 TI - Analysing the difference due to risk and demographic factors for incidence or mortality. AB - BACKGROUND: From an epidemiological and public health perspective there is an interest in quantifying differences in incidence and mortality between either time points, geographical areas or males and females. We propose a method for splitting such a difference in the number of cases/deaths into three components: (1) those due to risk; (2) those due to population structure (i.e. age distribution); and (3) those due to population size. We also propose graphical methods for presenting the results. Three examples are used to illustrate our methodology. PMID- 11034973 TI - Is audio computer-assisted self-interviewing a feasible method of surveying in Zimbabwe? AB - BACKGROUND: Research into reproductive health is dependent on participants accurately reporting sensitive behaviours. We examined whether audio computer assisted self-interviewing (ACASI), which increased sensitive behaviour reporting in the US, is a feasible method of surveying in developing countries. METHODS: Zimbabwean women in three educational groups were surveyed about demographics and family planning using interviewer and ACASI modes. An exit survey was administered to elicit information about the participants' opinions and experiences using ACASI. RESULTS: The majority of women (86%) preferred ACASI to interviewer mode. The reasons mentioned were always related to increased confidentiality and privacy. Ability to use ACASI and user preferences varied with educational level. More women with primary school or less education (53%) reported problems with computer use than women in the higher educational groups (10-12%). The percentage of women having perfect response concordance between ACASI and interviewer modes increased significantly with education (64%, 81%, and 84% respectively; P(trend) < 0.001). CONCLUSIONS: Use of ACASI may be more feasible in Zimbabwe and other developing countries than was originally thought, but ACASI programs should continue to be improved and tested in various countries and population groups. PMID- 11034974 TI - Use of comorbidity scores for control of confounding in studies using administrative databases. AB - BACKGROUND: Comorbidity scores are increasingly used to reduce potential confounding in epidemiological research. Our objective was to compare metrical and practical properties of published comorbidity scores for use in epidemiological research with administrative databases. METHODS: The literature was searched for studies of the validity of comorbidity scores as predictors of mortality and health service use, as measured by change in the area under the receiver operating characteristic (ROC) curve for dichotomous outcomes, and change in R(2) for continuous outcomes. RESULTS: Six scores were identified, including four versions of the Charlson Index (CI) which use either the three digit International Classification of Diseases, Ninth Revision (ICD-9) or the full ICD-9-CM (clinical modification) code, and two versions of the Chronic Disease Score (CDS) which used outpatient pharmacy records. Depending on the population and exposure under study, predictive validities varied between c = 0.64 and c = 0.77 for in-hospital or 30-day mortality. This is only a slight improvement over age adjustment. In one study the simple measure 'number of diagnoses' outperformed the CI (c = 0.73 versus c = 0.65). Proprietary scores like Ambulatory Diagnosis Groups and Patient Management Categories do not necessarily perform better in predicting mortality. Comorbidity indices are susceptible to a variety of coding errors. CONCLUSIONS: Comorbidity scores, particularly the CDS or D'Hoore's CI based on three-digit ICD-9 codes, may be useful in exploratory data analysis. However, residual confounding by comorbidity is inevitable, given how these scores are derived. How much residual confounding usually remains is something that future studies of comorbidity scores should examine. In any given study, better control for confounding can be achieved by deriving study-specific weights, to aggregate comorbidities into groups with similar relative risks of the outcomes of interest. PMID- 11034975 TI - Birds of a feather: using a rotational box plot to assess ascertainment bias. AB - BACKGROUND: Comparability of study participants with non-participants is customarily assessed by contrasting the distributions of sociodemographic characteristics. Such comparisons do not necessarily provide insight into whether or not participants of a given subgroup are similar to non-participants of the same subgroup. A geographical information system (GIS) may provide such insight by visually displaying the spatial distributions of participants and non participants. In a previously reported study of heterosexuals at elevated risk for human immunodeficiency virus (HIV), traditional methods suggested distributional differences in the demographic characteristics of participants and non-participants. METHODS: Based on residential address co-ordinates for each subgroup member, we used the subgroup's centroid as the origin and constructed a 360 degrees series of overlapping box plots of the distance of subgroups members to the origin, thereby producing closed polygons for each of the box plot demarcators. RESULTS: These rotational box plots revealed similar geographical distributions for most participant and non-participant subgroups, with the exception of African-American men and women. CONCLUSIONS: Observed differences resulted in part from the study design, and provided some insight into sampling problems encountered in social network studies. Based on Tobler's supposition that 'nearby things tend to be alike', the rotational box plot is a useful additional tool for investigating sample bias. PMID- 11034976 TI - Detection of epidemics in their early stage through infectious disease surveillance. AB - BACKGROUND: Surveillance of infectious diseases is done in many countries. The aims of such surveillance include the detection of epidemics. In the present study, the possibility of detecting an epidemic in its early stage using a simple method was evaluated for 16 infectious diseases. METHODS: We used as an index the number of cases per week per sentinel medical institution in the area covered by a health centre in infectious disease surveillance in Japan in 1993-1997. Periods of epidemics in health centre areas were determined according to the reported indices. The simple method used for detecting the early stage of an epidemic is that if the index exceeds a critical value, then an epidemic will begin in the following 4 weeks. The sensitivity, specificity and positive predictive value for this epidemic warning were evaluated for given critical values. RESULTS: When the specificity of the epidemic warning was more than 95%, the sensitivity was more than 60% in ten diseases, and more than 80% in four diseases (influenza-like illness, rubella, hand-foot-and-mouth disease, and herpangina). The positive predictive value was between 15.6% and 31.4% in these ten diseases. CONCLUSION: The early stage of epidemics of some infectious diseases might be detectable using this simple method. PMID- 11034977 TI - Alcohol and HIV: a study among sexually active adults in rural southwest Uganda. AB - OBJECTIVE: To investigate the association between alcohol consumption and HIV sero-positivity in a rural Ugandan population. METHODS: The adult population residing in a cluster of 15 neighbouring villages has been kept under epidemiological surveillance for HIV infection using annual censuses and sero surveys since 1989. At the eighth annual survey all respondents were asked about their history of alcohol consumption, the sale of alcohol in their household, and other socio-demographic information. After informed consent, blood was drawn for HIV serology. RESULTS: Of the total adult population 3279 (60%) were interviewed; 48% were males; 905 (27%) had not started sexual activity and were excluded from further analysis. Of the remaining 2374, 8% were HIV infected, 57% had ever drunk alcohol, and 4% lived in households where alcohol was sold. Living in a household where alcohol was sold was associated with a history of having ever drunk alcohol (OR 2.9, 95% CI : 1.7-4.8). HIV prevalence among adults living in households selling alcohol was 15% compared with 8% among those living in households not selling alcohol (OR 2.0, 95% CI : 1.1-3.6). Individuals who had ever drunk alcohol experienced an HIV prevalence twice that of those who had never drunk, 10% versus 5% (OR 2.0, 95% CI : 1.5-2.8). This association remained after adjusting for potential confounders including sale of alcohol in the household and Muslim religion (OR 1.8, 95% CI : 1.2-2.7). Only age, marital status and having ever drunk alcohol independently predicted HIV sero-positivity in a logistic regression model. CONCLUSIONS: We have demonstrated an association between a history of alcohol consumption and being HIV sero-positive. This unexplored factor may explain in part the observed lower prevalence of HIV infection among Muslims. Public health campaigns need to stress the relationship between HIV and alcohol. PMID- 11034978 TI - Association between clinical type of diarrhoea and growth of children under 5 years in rural Bangladesh. AB - BACKGROUND: The role of diarrhoea in the aetiology of growth retardation in young children remains controversial. To evaluate this, a population-based, longitudinal study of young children aged 6-48 months was conducted in Matlab, a rural area of Bangladesh, between May 1988 and April 1989. METHODS: Data obtained from 584 children were examined by one-year (n = 412) and 3-month (n = 1220) growth periods. Each growth period was analysed based on clinical types of diarrhoea, namely, non-diarrhoea, non-dysentery diarrhoea (diarrhoea without blood), and dysentery (diarrhoea with blood). Weight and height gains were compared among the study groups initially by one-way analysis of variance followed by multivariate analysis adjusting for potential confounding variables. RESULTS: Compared to non-diarrhoea and non-dysentery diarrhoea, dysentery was associated with significantly lower annual weight gain (1866 g [P < 0.01] and 1550 g [P < 0.05] versus 1350 g, respectively) and height gain (6.51 cm and 5.87 cm versus 5.27 cm [P < 0.01], respectively). Both 3-month dysentery and non dysentery intervals were significantly associated with less weight gain compared to non-diarrhoea intervals (490 g and 522 g versus 637 g [P: < 0.05], respectively). Dysentery intervals were also associated with significantly poorer height gain compared to other intervals (2.19 cm versus 2.42 cm [P < 0.05] and 2.46 cm [P < 0.01], respectively). CONCLUSIONS: The growth of young children is strongly influenced by the clinical type of diarrhoea and the impact is dependent on the proportion of dysentery episodes in the total diarrhoeal burden. PMID- 11034979 TI - Hepatitis C virus (HCV) infection and liver-related mortality: a population-based cohort study in southern Italy. The Association for the Study of Liver Disease in Puglia. AB - BACKGROUND: Hepatitis C virus (HCV) is a common cause of chronic liver diseases but the degree to which these diseases contribute to liver-related mortality is not well established. The aim of this study was to estimate the absolute and relative effects of HCV infection on liver-related mortality. METHODS: A population random sample of 2472 subjects aged > or = 30 years was enrolled and followed up from 1985 to 1996. At enrollment, a structured interview and a clinical evaluation were performed. Serum samples were tested using HCV ELISA and RIBA HCV. Outcomes were overall and liver-related mortality and tracing procedures included review of office and hospital records, death certificates, and interviews with general practitioners, attending hospital and next of kin. Statistical analysis was performed using Poisson and binomial prospective data regression. RESULTS: Crude overall and liver-related mortality rates were 7.66 (95% CI : 6.68-8.79) and 0.9 (95% CI : 0.3-2.2) per 10(3) person-years, respectively. For HCV infection effect, incidence rate ratio and difference (per 10(3) person-year), risk ratio and difference were 27.5 (95% CI : 6.5-115.6), 4 (95% CI : 3-7), 33.1 (95% CI : 7.8- 139.3) and 0.06 (95% CI : 0.04-0.08), respectively; all measures were adjusted for age at death, sex and daily alcohol intake. CONCLUSIONS: The results show a strong relative but weak absolute effect of HCV infection on liver-related mortality in the 10-year period considered. Poisson and binomial models are virtually equivalent, but the choice of the summarizing measure of effect may have a different impact on health policy. PMID- 11034980 TI - Seroepidemiology of Helicobacter pylori infection in a population of Egyptian children. AB - BACKGROUND: To describe the seroepidemiology of Helicobacter pylori infection in a population of Egyptian children under 3 years. METHODS: A cohort of children under 36 months, residing in Abu Homos, Egypt, were visited at home twice weekly. Information regarding the child's breastfeeding status was obtained, and periodic anthropometric and household hygiene surveys were performed. In June 1997, a serosurvey was conducted on 187 study participants over 6 months old. The serosurvey was repeated in October 1997. All sera were tested for IgG antibodies to H. pylori. RESULTS: The June prevalence of H. pylori infection was 10%, and the incidence from June to October was 15%. Between June and October, 8 (42%) of 19 children that were positive for H. pylori infection seroreverted to negative. All seroreversions occurred in children 6-17 months. Other than age, no sociodemographic or environmental factor was significantly associated with incident H. pylori infection. There was no significant differences in the weight for-age, weight-for-height, and height-for-age z-scores between children with and without prevalent H. pylori infection. CONCLUSIONS: Infection with H. pylori is common in Egyptian children under 3 years old and is not associated with malnutrition. No predictors for H. pylori infection were found. Our preliminary evidence for transient H. pylori infections in young children needs to be confirmed in a prospective cohort study, and predictors for persistent infection should be sought, since only these may be relevant to the known sequellae of infection. PMID- 11034981 TI - Effectiveness of incidence thresholds for detection and control of meningococcal meningitis epidemics in northern Togo. AB - BACKGROUND: Early outbreak detection is necessary for control of meningococcal meningitis epidemics. A weekly incidence of 15 cases per 100 000 inhabitants averaged over 2 consecutive weeks is recommended by the World Health Organization (WHO) for detection of meningitis epidemics in Africa. This and other thresholds are tested for ability to predict outbreaks and timeliness for control measures. METHODS: Meningitis cases recorded for 1990-1997 in health centres of northern Togo were reviewed. Weekly and annual incidences were determined for each district. Ability of different weekly incidence thresholds to detect outbreaks was assessed according to sensitivity, specificity, and positive and negative predictive values. The number of cases potentially prevented by reactive vaccination in 1997 was calculated for each threshold. RESULTS: Outbreaks occurred in 1995-1996 and in 1996-1997. The WHO-recommended threshold had good specificity but low sensitivity. Thresholds of 10 and 7 cases per 100,000 inhabitants in one week had sensitivity and specificity of 100% and increased the time available for intervention by more than one or two weeks, respectively. A maximum of 65% of cases could have been prevented during the 1997 epidemic, with up to 8% fewer cases prevented for each week of delay in achieving vaccine coverage. CONCLUSIONS: In northern Togo, thresholds of 7 or 10 cases per 100,000 inhabitants per week were excellent predictors of meningitis epidemics and allowed more time for a reactive vaccination strategy than current recommendations. PMID- 11034982 TI - High prevalence of congenital toxoplasmosis in Brazil estimated in a 3-year prospective neonatal screening study. AB - BACKGROUND: A pilot neonatal screening programme revealed a high (approximately 1 per 4800 live births) prevalence of congenital toxoplasmosis (CT) in the State of Rio Grande do Sul, Brazil. The purpose of this paper was to estimate in a larger prospective study the prevalence of CT in the country. METHODS: At the beginning of the study, an in-house indirect enzyme immunoassay (EIA) was used, to be later replaced with a commercial capture IgM fluorometric enzyme immunoassay (FEIA). Both methods detect specific anti-Toxoplasma gondii IgM-class antibodies eluted from dried blood spots. RESULTS: Of the total of 140,914 samples received from all over the country, 47 cases were identified and confirmed as CT. This finding suggests a prevalence of 1 per 3000 live births. Of the 47 patients, only eight (17%) had clinical manifestations: two had intracranial calcifications, four had retinal scars, one had an intracranial calcification and retinal scars, and one had hepatosplenomegaly with lymphoadenopathy. The testing was paid for by the patients' families who volunteered for the study and gave their informed consent. CONCLUSION: The 3-year prospective study using sensitive detection methods, reliable confirmation, and feedback from clinicians showed that CT has an extraordinarily high prevalence in Brazil, in fact the highest ever reported in the world. Although the long-term efficacy of treatment of CT has not been well documented, in view of the availability of reliable diagnostics, confirmation and monitoring, functional logistics, and networking for screening, the insidious nature of the sequelae and the very high prevalence of the disease, neonatal screening for CT should be considered an alternative to no screening at all. PMID- 11034983 TI - Long-term psychological stress and heart disease. PMID- 11034984 TI - The sex ratio of offspring of men exposed to metal fumes. PMID- 11034985 TI - International Labour Office: new study warns of HIV/AIDS catastrophe for workers and employers. PMID- 11034987 TI - Erratum PMID- 11034988 TI - Erratum PMID- 11034986 TI - New public/private sector effort initiated to accelerate access to HIV/AIDS care and treatment in developing countries. PMID- 11034989 TI - Structural basis for the impaired channeling and allosteric inter-subunit communication in the beta A169L/beta C170W mutant of tryptophan synthase. AB - We determined the 2.25 A resolution crystal structure of the betaA169L/betaC170W mutant form of the tryptophan synthase alpha(2)beta(2) complex from Salmonella typhimurium complexed with the alpha-active site substrate analogue 5-fluoro indole-propanol-phosphate to identify the structural basis for the changed kinetic properties of the mutant (Anderson, K. S., Kim, A. Y., Quillen, J. M., Sayers, E., Yang, X. J., and Miles, E. W. (1995) J. Biol. Chem. 270, 29936 29944). Comparison with the wild-type enzyme showed that the betaTrp(170) side chain occludes the tunnel connecting the alpha- and beta-active sites, explaining the accumulation of the intermediate indole during a single enzyme turnover. To prevent a steric clash between betaLeu(169) and betaGly(135), located in the beta sheet of the COMM (communication) domain (betaGly(102)-betaGly(189)), the latter reorganizes. The changed COMM domain conformation results in a loss of the hydrogen bonding networks between the alpha- and beta-active sites, explaining the poor activation of the alpha-reaction upon formation of the aminoacrylate complex at the beta-active site. The 100-fold reduced affinity for serine seems to result from a movement of betaAsp(305) away from the beta-active site so that it cannot interact with the hydroxyl group of a pyridoxal phosphate-bound serine. The proposed structural dissection of the effects of each single mutation in the betaA169L/betaC170W mutant would explain the very different kinetics of this mutant and betaC170F. PMID- 11034990 TI - An isoleucine-based allosteric switch controls affinity and shape shifting in integrin CD11b A-domain. AB - In response to cell activation signals, integrins switch from a low to a high affinity state. Physiologic ligands bind to integrins through a von Willebrand Factor A-type domain. Crystallographic studies revealed two conformations of this domain, "closed" and "open." The latter crystallizes in complex with a pseudoligand or ligand, suggesting that it represents the high affinity state; data linking structure and activity are lacking however. In this communication, we expressed stable low and high affinity forms of integrin CD11b A-domain and determined their binding isotherms and crystal structures. The low affinity form, generated by deleting an N-terminal extension extrinsic to the domain, did not bind to physiologic ligands, and crystallized in the closed conformation. The high affinity form was generated by either deleting or substituting an invariable C-terminal Ile(316), wedged into a hydrophobic socket in the closed form, but displaced from it in the open structure. Both mutants crystallized in the open conformation, and the Ile(316) --> Gly-modified integrin displayed high affinity. Structural differences between the low and high affinity forms were detected in solution. These data establish the structure-function correlates for the CD11b A domain, and define a ligand-independent isoleucine-based allosteric switch intrinsic to this domain that controls its conformation and affinity. PMID- 11034991 TI - The phosphatidylethanolamine-binding protein is the prototype of a novel family of serine protease inhibitors. AB - Serine proteases are involved in many processes in the nervous system and specific inhibitors tightly control their proteolytic activity. Thrombin is thought to play a role in tissue development and homeostasis. To date, protease nexin-1 is the only known endogenous protease inhibitor that specifically interferes with thrombotic activity and is expressed in the brain. In this study, we report the detection of a novel thrombin inhibitory activity in the brain of protease nexin-1(-/-) mice. Purification and subsequent analysis by tandem mass spectrometry identified this protein as the phosphatidylethanolamine-binding protein (PEBP). We demonstrate that PEBP exerts inhibitory activity against several serine proteases including thrombin, neuropsin, and chymotrypsin, whereas trypsin, tissue type plasminogen activator, and elastase are not affected. Since PEBP does not share significant homology with other serine protease inhibitors, our results define it as the prototype of a novel class of serine protease inhibitors. PEBP immunoreactivity is found on the surface of Rat-1 fibroblast cells and although its sequence contains no secretion signal, PEBP-H(6) can be purified from the conditioned medium upon recombinant expression. PMID- 11034992 TI - Structure and site-directed mutagenesis of a flavoprotein from Escherichia coli that reduces nitrocompounds: alteration of pyridine nucleotide binding by a single amino acid substitution. AB - The crystal structure of a major oxygen-insensitive nitroreductase (NfsA) from Escherichia coli has been solved by the molecular replacement method at 1.7-A resolution. This enzyme is a homodimeric flavoprotein with one FMN cofactor per monomer and catalyzes reduction of nitrocompounds using NADPH. The structure exhibits an alpha + beta-fold, and is comprised of a central domain and an excursion domain. The overall structure of NfsA is similar to the NADPH-dependent flavin reductase of Vibrio harveyi, despite definite difference in the spatial arrangement of residues around the putative substrate-binding site. On the basis of the crystal structure of NfsA and its alignment with the V. harveyi flavin reductase and the NADPH-dependent nitro/flavin reductase of Bacillus subtilis, residues Arg(203) and Arg(208) of the loop region between helices I and J in the vicinity of the catalytic center FMN is predicted as a determinant for NADPH binding. The R203A mutant results in a 33-fold increase in the K(m) value for NADPH indicating that the side chain of Arg(203) plays a key role in binding NADPH possibly to interact with the 2'-phosphate group. PMID- 11034993 TI - Characterization of intercellular adhesion molecule-1 regulation by Epstein-Barr virus-encoded latent membrane protein-1 identifies pathways that cooperate with nuclear factor kappa B to activate transcription. AB - The latent membrane protein-1 (LMP1) of Epstein-Barr virus induces gene transcription, phenotypic changes, and oncogenic transformation. One cellular gene induced by LMP1 is that for intercellular adhesion molecule-1 (ICAM-1), which participates in a wide range of inflammatory and immune responses. ICAM-1 may enhance the immune recognition of cells transformed by Epstein-Barr virus, and thus combat development of malignancy. Despite growing understanding of the various signaling functions of LMP1, the molecular mechanisms by which LMP1 induces ICAM-1 are not understood. Here, we demonstrate that transcriptional activation by LMP1 is absolutely dependent upon a variant NF-kappaB motif within the tumor necrosis factor alpha (TNFalpha) response element of the ICAM-1 promoter. Although the TNFalpha response element is sufficient for TNFalpha induction of the ICAM-1 promoter, LMP1 also required the cooperation of additional upstream sequences for optimal induction. Inhibitor studies of known LMP1-induced signaling pathways ruled out the involvement of c-Jun N-terminal kinase (JNK), p38 mitogen-activated protein kinase, and the Janus-activating tyrosine kinase 3 (JAK3), and confirmed NF-kappaB as a critical factor for induction of ICAM-1. However, although constitutive activation of NF-kappaB efficiently induced promoter activity, it was not sufficient to induce either ICAM-1 mRNA or ICAM-1 protein. Using signaling defective LMP1 mutants and deacetylation inhibitors, we showed that the C-terminal activator region 1 of LMP1 delivers a new cooperating signal to induce ICAM-1 mRNA. PMID- 11034995 TI - Distinct organization of DNA complexes of various HMGI/Y family proteins and their modulation upon mitotic phosphorylation. AB - High mobility group (HMG) proteins HMGI, HMGY, HMGI-C, and Chironomus HMGI are DNA-binding proteins thought to modulate the assembly and the function of transcriptional complexes. Each of these proteins contains three DNA-binding domains (DBD), properties of which appear to be regulated by phosphorylation. High levels of these proteins are characteristic for rapidly dividing cells in embryonic tissues and tumors. On the basis of their occurrence, specific functions for each of these proteins have been postulated. In this study we demonstrate differences in the nature of contacts of these proteins with promoter region of the interferon-beta gene. We show that HMGI and HMGY interact with this DNA via three DBDs, whereas HMGI-C and Chironomus HMGI bind to this DNA using only two domains. Phosphorylation of HMGY protein by Cdc2 kinase leads to impairing of contacts between the N-terminally located DBD and a single promoter element. The perturbations in the architecture of the protein.DNA complexes involve changes in the degree of unbending of the intrinsically bent IFNbeta promoter. Our results provide first insights into the molecular basis of functional specificity of proteins of the HMGI/Y family and their regulation by phosphorylation. PMID- 11034994 TI - Helicobacter pylori-induced prostaglandin E(2) synthesis involves activation of cytosolic phospholipase A(2) in epithelial cells. AB - Helicobacter pylori initiates an inflammatory response and gastric diseases, which are more common in patients infected with H. pylori strains carrying the pathogenicity island, by colonizing the gastric epithelium. In the present study we investigated the mechanism of prostaglandin E(2) (PGE(2)) synthesis in response to H. pylori infection. We demonstrate that H. pylori induces the synthesis of PGE(2) via release of arachidonic acid predominately from phosphatidylinositol. In contrast to H. pylori wild type, an isogenic H. pylori strain with a mutation in the pathogenicity island exerts only weak arachidonic acid and PGE(2) synthesis. The H. pylori-induced arachidonic acid release was abolished by phospholipase A(2) (PLA(2)) inhibitors and by pertussis toxin (affects the activity of G alpha(i)/G alpha(o)). The role of phospholipase C, diacylglycerol lipase, or phospholipase D was excluded by using specific inhibitors. An inhibitor of the stress-activated p38 kinase (SB202190), but neither inhibitors of protein kinase C nor an inhibitor of the extracellular regulated kinase pathway (PD98059), decreased the H. pylori-induced arachidonic acid release. H. pylori-induced phosphorylation of p38 kinase and cytosolic PLA(2) was blocked by SB202190. These results indicate that H. pylori induces the release of PGE(2) from epithelial cells by cytosolic PLA(2) activation via G alpha(i)/G alpha(o) proteins and the p38 kinase pathway. PMID- 11034996 TI - Oxidized phospholipids induce changes in hepatic paraoxonase and ApoJ but not monocyte chemoattractant protein-1 via interleukin-6. AB - In this study, we tested if interleukin-6 (IL-6) plays a role in mediating the effects of oxidized phospholipids (OXPL). Treatment of HepG2 cells with oxidized 1-palmitoyl-2-arachidonyl-sn-glycero-3-phosphoryl choline (OX-PAPC), or biologically active lipids present in mildly oxidized low density lipoprotein, increased apolipoprotein J (apoJ), and decreased paraoxonase (PON) mRNA levels. Antibodies to IL-6 blocked these changes. IL-6 treatment in the absence of OXPL produced the same pattern of mRNA changes observed with OXPL treatment alone. In vivo, OX-PAPC injected into C57BL/6J mice resulted in a marked reduction in PON activity and an increase in apoJ levels in plasma after 16 h. Injection of OX PAPC into IL-6-deficient C57BL/6J mice (IL-6 -/-) did not alter either PON activity or apoJ levels. We then tested if other mechanisms involved in fatty streak formation depended upon IL-6. Antibody to IL-6 had no effect on OX-PAPC induced secretion of MCP-1 by endothelial cells nor on MCP-1 mRNA expression in HepG2 cells. C57BL/6J and IL-6 -/- mice fed an atherogenic diet both demonstrated markedly reduced plasma PON activities and the IL-6 -/- mice developed fatty streaks to a greater degree than wild-type mice. We conclude that IL-6 is critical to short term but not long term regulation of PON and that IL-6 is not required for OXPL regulation of MCP-1. PMID- 11034997 TI - Hyperoxia inhibits oxidant-induced apoptosis in lung epithelial cells. AB - It has previously been shown that hyperoxia induces nonapoptotic cell death in cultured lung epithelial cells, whereas hydrogen peroxide (H(2)O(2)) and paraquat cause apoptosis. To test whether pathways leading to oxidative apoptosis in epithelial cells are sensitive to molecular O(2), A549 cells were exposed to 95% O(2) prior to exposure to lethal concentrations of H(2)O(2). The extent of H(2)O(2)-induced apoptosis was significantly reduced in cells preexposed to hyperoxia compared with room-air controls. Preexposure of the hyperoxia-resistant HeLa-80 cell line to 80% O(2) also inhibited oxidant-induced apoptosis, suggesting that this inhibition is not due to O(2) toxicity. Because hyperoxia generates reactive oxygen species and activates the redox-sensitive transcription factor nuclear factor kappa B (NF-kappa B), the role of antioxidant enzymes and NF-kappa B were examined in this inhibitory process. The onset of inhibition appeared to be directly related to the degradation of I kappa B and subsequent activation of NF-kappa B (either by hyperoxia or TNF-alpha), whereas no significant up-regulation of endogenous antioxidant enzyme activities was found. In addition, suppression of NF-kappa B activities by transfecting A549 cells with a dominant-negative mutant construct of I kappa B significantly augmented the extent of H(2)O(2)-induced apoptosis. These data suggest that hyperoxia inhibits oxidant-induced apoptosis and that this inhibition is mediated by NF-kappa B. PMID- 11034998 TI - Characterization of the intramolecular electron transfer pathway from 2 hydroxyphenazine to the heterodisulfide reductase from Methanosarcina thermophila. AB - Heterodisulfide reductase (HDR) is a component of the energy-conserving electron transfer system in methanogens. HDR catalyzes the two-electron reduction of coenzyme B-S-S-coenzyme M (CoB-S-S-CoM), the heterodisulfide product of the methyl-CoM reductase reaction, to free thiols, HS-CoB and HS-CoM. HDR from Methanosarcina thermophila contains two b-hemes and two [Fe(4)S(4)] clusters. The physiological electron donor for HDR appears to be methanophenazine (MPhen), a membrane-bound cofactor, which can be replaced by a water-soluble analog, 2 hydroxyphenazine (HPhen). This report describes the electron transfer pathway from reduced HPhen (HPhenH(2)) to CoB-S-S-CoM. Steady-state kinetic studies indicate a ping-pong mechanism for heterodisulfide reduction by HPhenH(2) with the following values: k(cat) = 74 s(-1) at 25 degrees C, K(m) (HPhenH(2)) = 92 microm, K(m) (CoB-S-S-CoM) = 144 microm. Rapid freeze-quench EPR and stopped-flow kinetic studies and inhibition experiments using CO and diphenylene iodonium indicate that only the low spin heme and the high potential FeS cluster are involved in CoB-S-S-CoM reduction by HPhenH(2). Fe-S cluster disruption by mersalyl acid inhibits heme reduction by HPhenH(2), suggesting that a 4Fe cluster is the initial electron acceptor from HPhenH(2). We propose the following electron transfer pathway: HPhenH(2) to the high potential 4Fe cluster, to the low potential heme, and finally, to CoB-S-S-CoM. PMID- 11034999 TI - Regulation of DNA binding and trans-activation by a xenobiotic stress-activated plant transcription factor. AB - As-1-type cis-elements augment transcription of both nuclear and pathogen genes in response to stress and defense cues in plants. Basic/leucine zipper proteins termed "TGA factors" that specifically bind as-1 elements are likely candidates for mediating these transcription activities. Our earlier work has shown that 2, 4-dichlorophenoxyacetic acid-induced xenobiotic stress enhances trans-activation by a chimeric fusion protein of the yeast Gal4 binding domain and TGA1a, a TGA factor of tobacco. Here we demonstrate that xenobiotic stress also enhances the ability of native TGA1a to bind as-1 and activate transcription of a known target gene. In addition, the previously identified xenobiotic stress-responsive domain of TGA1a was found to inhibit this factor's trans-activation potential by a mechanism that appears to involve stimulus-reversible interactions with a nuclear repressor protein. Results from these and other studies can now be placed in the context of a working model to explain basal and xenobiotic stress-induced activities of TGA1a through its cognate cis-acting element. PMID- 11035001 TI - Smooth muscle differentiation marker gene expression is regulated by RhoA mediated actin polymerization. AB - Smooth muscle cell (SMC) differentiation is regulated by a complex array of local environmental cues, but the intracellular signaling pathways and the transcription mechanisms that regulate this process are largely unknown. We and others have shown that serum response factor (SRF) contributes to SMC-specific gene transcription, and because the small GTPase RhoA has been shown to regulate SRF, the goal of the present study was to test the hypothesis that RhoA signaling is a critical mechanism for regulating SMC differentiation. Coexpression of constitutively active RhoA in rat aortic SMC cultures significantly increased the activity of the SMC-specific promoters, SM22 and SM alpha-actin, whereas coexpression of C3 transferase abolished the activity of these promoters. Inhibition of either stress fiber formation with the Rho kinase inhibitor Y-27632 (10 microm) or actin polymerization with latrunculin B (0.5 microm) significantly decreased the activity of SM22 and SM alpha-actin promoters. In contrast, increasing actin polymerization with jasplakinolide (0.5 microm) increased SM22 and SM alpha-actin promoter activity by 22-fold and 13-fold, respectively. The above interventions had little or no effect on the transcription of an SRF dependent c-fos promoter or on a minimal thymidine kinase promoter that is not SRF-dependent. Taken together, the results of these studies indicate that in SMC, RhoA-dependent regulation of the actin cytoskeleton selectively regulates SMC differentiation marker gene expression by modulating SRF-dependent transcription. The results also suggest that RhoA signaling may serve as a convergence point for the multiple signaling pathways that regulate SMC differentiation. PMID- 11035002 TI - Helix packing of functionally important regions of the cardiac Na(+)-Ca(2+) exchanger. AB - In a revised topological model of the cardiac Na(+)-Ca(2+) exchanger, there are nine transmembrane segments (TMSs) and two possible re-entrant loops (Nicoll, D. A., Ottolia, M., Lu, Y., Lu, L., and Philipson, K. D. (1999) J. Biol. Chem. 274, 910-917; Iwamoto, T., Nakamura, T. Y., Pan, Y., Uehara, A., Imanaga, I., and Shigekawa, M. (1999) FEBS Lett. 446, 264-268). The TMSs form two clusters separated by a large intracellular loop between TMS5 and TMS6. We have combined cysteine mutagenesis and oxidative cross-linking to study proximity relationships of TMSs in the exchanger. Pairs of cysteines were reintroduced into a cysteine less exchanger, one in a TMS in the NH(2)-terminal cluster (TMSs 1-5) and the other in a TMS in the COOH-terminal cluster (TMSs 6-9). The mutant exchanger proteins were expressed in HEK293 cells, and disulfide bond formation between introduced cysteines was analyzed by gel mobility shifts. Western blots showed that S117C/V804C, A122C/Y892C, A151C/T815C, and A151C/A821C mutant proteins migrated at 120 kDa under reducing conditions and displayed a partial mobility shift to 160 kDa under nonreducing conditions. This shift indicates the formation of a disulfide bond between these paired cysteine residues. Copper phenanthroline and the cross-linker N', N'-o-phenylenedimaleimide enhanced the mobility shift to 160 kDa. Our data suggest that TMS7 is close to TMS3 near the intracellular side of the membrane and is in the vicinity of TMS2 near the extracellular surface. Also, TMS2 must adjoin TMS8. This initial packing model of the exchanger brings two functionally important domains in the exchanger, the alpha 1 and alpha 2 repeats, close to each other. PMID- 11035003 TI - Pores formed by single subunits in mixed dimers of different CLC chloride channels. AB - CLC chloride channels comprise a gene family with nine mammalian members. Probably all CLC channels form homodimers, and some CLC proteins may also associate to heterodimers. ClC-0 and ClC-1, the only CLC channels investigated at the single-channel level, display two conductances of equal size which are thought to result from two separate pores, formed individually by the two monomers. We generated concatemeric channels containing one subunit of ClC-0 together with one subunit of ClC-1 or ClC-2. They should display two different conductances if one monomer were sufficient to form one pore. Indeed, we found a 8-picosiemens (pS) conductance (corresponding to ClC-0) that was associated with either a 1.8-pS (ClC-1) or a 2.8-pS (ClC-2) conductance. These conductances retained their typical gating, but the slow gating of ClC-0 that affects both pores simultaneously was lost. ClC-2 and ClC-0 current components were modified by point mutations in the corresponding subunit. The ClC-2 single pore of the mixed dimer was compared with the pores in the ClC-2 homodimer and found to be unaltered. We conclude that each monomer individually forms a gated pore. CLC dimers in general must be imagined as having two pores, as shown previously for ClC-0. PMID- 11035004 TI - C-terminal elements control location, activation threshold, and p38 docking of ribosomal S6 kinase B (RSKB). AB - RSKB, a p90 ribosomal S6 protein kinase with two catalytic domains, is activated by p38- and extracellular signal-regulated kinase mitogen-activated protein kinase pathways. The sequences between the two catalytic domains and of the C terminal extension contain elements that control RSKB activity. The C-terminal extension of RSKB presents a putative bipartite (713)KRX(14)KRRKQKLRS(737) nuclear location signal. The distinct cytoplasmic and nuclear locations of various C-terminal truncation mutants supported the hypothesis that the nuclear location signal was essential to direct RSKB to the nuclear compartment. The (725)APLAKRRKQKLRS(737) sequence also was essential for the intermolecular association of RSKB with p38. The activation of RSKB through p38 could be dissociated from p38 docking, because RSKB truncated at Ser(681) strongly responded to p38 pathway activity. Interestingly, Delta(725-772)-RSKB was nearly nonresponsive to p38. Sequence alignment with the autoinhibitory C-terminal extension of Ca+2/calmodulin-dependent protein kinase I predicted a conserved regulatory (708)AFN(710) motif. Alanine mutation of the key Phe709 residue resulted in strongly elevated basal level RSKB activity. A regulatory role also was assigned to Thr687, which is located in a mitogen-activated protein kinase phosphorylation consensus site. These findings support that the RSKB C-terminal extension contains elements that control activation threshold, subcellular location, and p38 docking. PMID- 11035005 TI - c-Src regulates the interaction between connexin-43 and ZO-1 in cardiac myocytes. AB - Connexin-43 is known to interact directly with ZO-1 in cardiac myocytes, but little is known about the role of ZO-1 in connexin-43 function. In cardiac myocytes, constitutively active c-Src inhibited endogenous interaction between connexin-43 and ZO-1 by binding to connexin-43. In HEK293 cells, by contrast, a connexin-43 mutant lacking the Src phosphorylation site (Tyr265) interacted with ZO-1 despite cotransfection of a constitutively active c-Src. Moreover, in vitro binding assays using recombinant proteins synthesized from regions of connexin-43 and ZO-1 showed that the tyrosine-phosphorylated C terminus of connexin-43 interacts with the c-Src SH2 domain in parallel with the loss of its interaction with ZO-1. Cell surface biotinylation revealed that, by phosphorylating Tyr265, constitutively active c-Src reduces total and cell surface connexin-43 down to the levels seen in cells expressing a mutant connexin-43 lacking the ZO-1 binding domain. Finally, electrophysiological analysis showed that both the tyrosine phosphorylation site and the ZO-1-binding domain of connexin-43 were involved in the regulation of gap junctional function. We therefore conclude that c-Src regulates the interaction between connexin-43 and ZO-1 through tyrosine phosphorylation and through the binding of its SH2 domain to connexin-43. PMID- 11035006 TI - Induction of duplex to G-quadruplex transition in the c-myc promoter region by a small molecule. AB - A major control element of the human c-myc oncogene is the nuclease hypersensitive purine/pyrimidine-rich sequence. This double-stranded DNA fragment, corresponding to the 27-base pair segment in the nuclease hypersensitive element of the c-myc promoter region, forms a stable Watson-Crick double helix under physiological conditions. However, this duplex DNA can be effectively converted to G-quadruplex DNA by a small molecular weight ligand. Both intermolecular and intramolecular G-quadruplex forms can be induced by this ligand. Similar transitional changes are also observed with the duplex telomeric sequence from the Oxytricha species. These results provide additional support to the idea that G-quadruplex structures may play structural roles in vivo and also provide insight into novel methodologies for rational drug design. These structurally altered DNA elements might serve as regulatory signals in gene expression or in telomere dynamics and hence are promising targets for drug action. PMID- 11035007 TI - A novel gamma -secretase assay based on detection of the putative C-terminal fragment-gamma of amyloid beta protein precursor. AB - Alzheimer's disease is characterized by the deposits of the 4-kDa amyloid beta peptide (A beta). The A beta protein precursor (APP) is cleaved by beta-secretase to generate a C-terminal fragment, CTF beta, which in turn is cleaved by gamma secretase to generate A beta. Alternative cleavage of the APP by alpha-secretase at A beta 16/17 generates the C-terminal fragment, CTFalpha. In addition to A beta, endoproteolytic cleavage of CTF alpha and CTF beta by gamma-secretase should yield a C-terminal fragment of 57-59 residues (CTF gamma). However, CTF gamma has not yet been reported in either brain or cell lysates, presumably due to its instability in vivo. We detected the in vitro generation of A beta as well as an approximately 6-kDa fragment from guinea pig brain membranes. We have provided biochemical and pharmacological evidence that this 6-kDa fragment is the elusive CTF gamma, and we describe an in vitro assay for gamma-secretase activity. The fragment migrates with a synthetic peptide corresponding to the 57 residue CTF gamma fragment. Three compounds previously identified as gamma secretase inhibitors, pepstatin-A, MG132, and a substrate-based difluoroketone (t butoxycarbonyl-Val-Ile-(S)-4-amino-3-oxo-2, 2-difluoropentanoyl-Val-Ile-OMe), reduced the yield of CTF gamma, providing additional evidence that the fragment arises from gamma-secretase cleavage. Consistent with reports that presenilins are the elusive gamma-secretases, subcellular fractionation studies showed that presenilin-1, CTF alpha, and CTF beta are enriched in the CTF gamma-generating fractions. The in vitro gamma-secretase assay described here will be useful for the detailed characterization of the enzyme and to screen for gamma-secretase inhibitors. PMID- 11035009 TI - Characterization of the mRNA capping apparatus of Candida albicans. AB - The mRNA capping apparatus of the pathogenic fungus Candida albicans consists of three components: a 520- amino acid RNA triphosphatase (CaCet1p), a 449-amino acid RNA guanylyltransferase (Cgt1p), and a 474-amino acid RNA (guanine-N7-) methyltransferase (Ccm1p). The fungal guanylyltransferase and methyltransferase are structurally similar to their mammalian counterparts, whereas the fungal triphosphatase is mechanistically and structurally unrelated to the triphosphatase of mammals. Hence, the triphosphatase is an attractive antifungal target. Here we identify a biologically active C-terminal domain of CaCet1p from residues 202 to 520. We find that CaCet1p function in vivo requires the segment from residues 202 to 256 immediately flanking the catalytic domain from 257 to 520. Genetic suppression data implicate the essential flanking segment in the binding of CaCet1p to the fungal guanylyltransferase. Deletion analysis of the Candida guanylyltransferase demarcates an N-terminal domain, Cgt1(1-387)p, that suffices for catalytic activity in vitro and for cell growth. An even smaller domain, Cgt1(1-367)p, suffices for binding to the guanylyltransferase docking site on yeast RNA triphosphatase. Deletion analysis of the cap methyltransferase identifies a C-terminal domain, Ccm1(137-474)p, as being sufficient for cap methyltransferase function in vivo and in vitro. Ccm1(137-474)p binds in vitro to synthetic peptides comprising the phosphorylated C-terminal domain of the largest subunit of RNA polymerase II. Binding is enhanced when the C-terminal domain is phosphorylated on both Ser-2 and Ser-5 of the YSPTSPS heptad repeat. We show that the entire three-component Saccharomyces cerevisiae capping apparatus can be replaced by C. albicans enzymes. Isogenic yeast cells expressing "all-Candida" versus "all-mammalian" capping components can be used to screen for cytotoxic agents that specifically target the fungal capping enzymes. PMID- 11035010 TI - Mapping of the ATP-binding sites on inositol 1,4,5-trisphosphate receptor type 1 and type 3 homotetramers by controlled proteolysis and photoaffinity labeling. AB - Submillimolar ATP concentrations strongly enhance the inositol 1,4,5 trisphosphate (IP(3))-induced Ca(2+) release, by binding specifically to ATP binding sites on the IP(3) receptor (IP(3)R). To locate those ATP-binding sites on IP(3)R1 and IP(3)R3, both proteins were expressed in Sf9 insect cells and covalently labeled with 8-azido-[alpha-(32)P]ATP. IP(3)R1 and IP(3)R3 were then purified and subjected to a controlled proteolysis, and the labeled proteolytic fragments were identified by site-specific antibodies. Two fragments of IP(3)R1 were labeled, each containing one of the previously proposed ATP-binding sites with amino acid sequence GXGXXG (amino acids 1773-1780 and 2016-2021, respectively). In IP(3)R3, only one fragment was labeled. This fragment contained the GXGXXG sequence (amino acids 1920-1925), which is conserved in the three IP(3)R isoforms. The presence of multiple interaction sites for ATP was also evident from the IP(3)-induced Ca(2+) release in permeabilized A7r5 cells, which depended on ATP over a very broad concentration range from micromolar to millimolar. PMID- 11035011 TI - The single pore residue Asp542 determines Ca2+ permeation and Mg2+ block of the epithelial Ca2+ channel. AB - The epithelial Ca(2+) channel (ECaC), which was recently cloned from rabbit kidney, exhibits distinctive properties that support a facilitating role in transcellular Ca(2+) (re)absorption. ECaC is structurally related to the family of six transmembrane-spanning ion channels with a pore-forming region between S5 and S6. Using point mutants of the conserved negatively charged amino acids present in the putative pore, we have identified a single aspartate residue that determines Ca(2+) permeation of ECaC and modulation by extracellular Mg(2+). Mutation of the aspartate residue, D542A, abolishes Ca(2+) permeation and Ca(2+) dependent current decay as well as block by extracellular Mg(2+), whereas monovalent cations still permeate the mutant channel. Variation of the side chain length in mutations D542N, D542E, and D542M attenuated Ca(2+) permeability and Ca(2+)-dependent current decay. Block of monovalent currents through ECaC by Mg(2+) was decreased. Exchanging the aspartate residue for a positively charged amino acid, D542K, resulted in a nonfunctional channel. Mutations of two neighboring negatively charged residues, i.e. Glu(535) and Asp(550), had only minor effects on Ca(2+) permeation properties. PMID- 11035012 TI - Identification of a carboxyl-terminal diaphanous-related formin homology protein autoregulatory domain. AB - Mammalian and fungal Diaphanous-related formin homology (DRF) proteins contain several regions of conserved sequence homology. These include an amino-terminal GTPase binding domain (GBD) that interacts with activated Rho family members and formin homology domains that mediate targeting or interactions with signaling kinases and actin-binding proteins. DRFs also contain a conserved Dia autoregulatory domain (DAD) in their carboxyl termini that binds the GBD. The GBD is a bifunctional autoinhibitory domain that is regulated by activated Rho. Expression of the isolated DAD in cells causes actin fiber formation and stimulates serum response factor-regulated gene expression. Inhibitor experiments show that the effects of exogenous DAD expression are dependent upon cellular Dia proteins. Alanine substitution of DAD consensus residues that disrupt GBD binding also eliminate DAD biological activity. Thus, DAD expression activates nuclear signaling and actin remodeling by mimicking activated Rho and unlatching the autoinhibited state of the cellular complement of Dia proteins. PMID- 11035013 TI - Cd36, a member of the class b scavenger receptor family, as a receptor for advanced glycation end products. AB - Interaction of advanced glycation end products (AGE) with AGE receptors induces several cellular phenomena potentially relating to diabetic complications. Five AGE receptors identified so far are RAGE (receptor for AGE), galectin-3, 80K-H, OST-48, and SRA (macrophage scavenger receptor class A types I and II). Since SRA is known to belong to the class A scavenger receptor family, and the scavenger receptor collectively represents a family of multiligand lipoprotein receptors, it is possible that CD36, although belonging to the class B scavenger receptor family, can recognize AGE proteins as ligands. This was tested at the cellular level in this study using Chinese hamster ovary (CHO) cells overexpressing human CD36 (CD36-CHO cells). Cellular expression of CD36 was confirmed by immunoblotting and immunofluorescent microscopy using anti-CD36 antibody. Upon incubation at 37 degrees C, (125)I-AGE-bovine serum albumin (AGE-BSA) and (125)I oxidized low density lipoprotein (LDL), an authentic ligand for CD36, were endocytosed in a dose-dependent fashion and underwent lysosomal degradation by CD36-CHO cells, but not wild-type CHO cells. In binding experiments at 4 degrees C, (125)I-AGE-BSA exhibited specific and saturable binding to CD36-CHO cells (K(d) = 5.6 microg/ml). The endocytic uptake of (125)I-AGE-BSA by these cells was inhibited by 50% by oxidized LDL and by 60% by FA6-152, an anti-CD36 antibody inhibiting cellular binding of oxidized LDL. Our results indicate that CD36 expressed by these cells mediates the endocytic uptake and subsequent intracellular degradation of AGE proteins. Since CD36 is one of the major oxidized LDL receptors and is up-regulated in macrophage- and smooth muscle cell derived foam cells in human atherosclerotic lesions, these results suggest that, like oxidized LDL, AGE proteins generated in situ are recognized by CD36, which might contribute to the pathogenesis of diabetic macrovascular complications. PMID- 11035014 TI - The Rela(p65) subunit of NF-kappaB is essential for inhibiting double-stranded RNA-induced cytotoxicity. AB - Double-stranded RNA (dsRNA) molecules generated during virus infection can initiate a host antiviral response to limit further infection. Such a response involves induction of antiviral gene expression by the dsRNA-activated protein kinase (PKR) and the NF-kappaB transcription factor. In addition, dsRNA can also induce apoptosis by an incompletely understood mechanism that may serve to further limit viral replication. Here we demonstrate a novel role for the RelA subunit of NF-kappaB in inhibiting dsRNA-induced cell death. dsRNA treatment resulted in caspase 3 activation and apoptotic morphological transformations in mouse embryonic fibroblasts (MEFs) derived from RelA-/- mice but not from RelA+/+ mice. Such dsRNA-induced killing could be inhibited by expression of either a dominant-negative mutant of PKR or wild-type RelA. Interestingly, caspase 3 activated following dsRNA treatment of RelA-/- MEFs was essential for apoptotic nuclear changes but dispensable for cytotoxicity. A broader specificity caspase inhibitor was also unable to inhibit dsRNA-induced cytotoxicity, suggesting that caspase activation is not essential for the induction of cell death by dsRNA in MEFs. However, combined inhibition of caspase 3 and reactive oxygen species production resulted in complete inhibition of dsRNA-induced cytotoxicity. These results demonstrate an essential role for NF-kappaB in protecting cells from dsRNA-induced apoptosis and suggest that NF-kappaB may inhibit both caspase dependent and reactive oxygen species-dependent cytotoxic pathways. PMID- 11035015 TI - The role of endocytosis in regulating L1-mediated adhesion. AB - L1 is a neural cell adhesion molecule critical for neural development. Full length L1 (L1(FL)) contains an alternatively spliced cytoplasmic sequence, RSLE, which is absent in L1 expressed in nonneuronal cells. The RSLE sequence follows a tyrosine, creating an endocytic motif that allows rapid internalization via clathrin-mediated endocytosis. We hypothesized that L1(FL) would internalize more rapidly than L1 lacking the RSLE sequence (L1(Delta)(RSLE)) and that internalization might regulate L1-mediated adhesion. L1 internalization was measured by immunofluorescence microscopy and by uptake of (125)I-anti-rat-L1 antibody, demonstrating that L1(FL) is internalized 2-3 times faster than L1(Delta)(RSLE). Inhibition of clathrin-mediated endocytosis slowed internalization of L1(FL) but did not affect initial uptake of L1(Delta)(RSLE). To test whether L1 endocytosis regulates L1 adhesion, cell aggregation rates were tested. L1(Delta)(RSLE) cells aggregated two times faster than L1(FL) cells. Inhibition of clathrin-mediated endocytosis increases the aggregation rate of the L1(FL) cells to that of L1(Delta)(RSLE) cells. Our results demonstrate that rapid internalization of L1 dramatically affects L1 adhesion. PMID- 11035017 TI - Binding of the transcription effector ppGpp to Escherichia coli RNA polymerase is allosteric, modular, and occurs near the N terminus of the beta'-subunit. AB - Among the prokaryotae, the nucleotide ppGpp is a second messenger of physiological stress and starvation. The target of ppGpp is RNA polymerase, where it putatively binds and alters the enzyme's activity. Previous data had implicated the beta-subunit of Escherichia coli RNA polymerase as containing a single ppGpp binding site. In this study, a photocross-linkable derivative of ppGpp, 6-thioguanosine-3',5'-(bis)pyrophosphate (6-thio-ppGpp), was used to localize the ppGpp binding site. In in vitro transcription assays, 6-thio-ppGpp inhibited transcription from the argT promoter identically to bona fide ppGpp. The thio group of 6-thio-ppGpp is directly photoactivatable and is thus a zero length cross-linker. Cross-linking of RNA polymerase was directed primarily to the beta'-subunit and could be competed efficiently by native ppGpp but not by GTP or GDP. Cyanogen bromide digestion analysis of the cross-linked beta'-subunit was consistent with an extreme N-terminal cross-link. To assess allosteric consequences of ppGpp binding to RNA polymerase, high level trypsin resistance in the presence and absence of ppGpp was monitored. Trypsin digestion of RNA polymerase bound to ppGpp leads to protection of an N-terminal fragment of the beta'-subunit and a C-terminal fragment of the beta-subunit. We propose that the N terminus of beta' together with the C terminus of beta constitute a modular ppGpp binding site. PMID- 11035016 TI - A new family of Cdc42 effector proteins, CEPs, function in fibroblast and epithelial cell shape changes. AB - Cdc42, a Rho GTPase, regulates the organization of the actin cytoskeleton by its interaction with several distinct families of downstream effector proteins. Here, we report the identification of four new Cdc42-binding proteins that, along with MSE55, constitute a new family of effector proteins. These molecules, designated CEPs, contain three regions of homology, including a Cdc42 binding domain and two unique domains called CI and CII. Experimentally, we have verified that CEP2 and CEP5 bind Cdc42. Expression of CEP2, CEP3, CEP4, and CEP5 in NIH-3T3 fibroblasts induced pseudopodia formation. Fibroblasts coexpressing dominant negative Cdc42 with CEP2 or expressing a Cdc42/Rac interactive binding domain mutant of CEP2 did not induce pseudopodia formation. In primary keratinocytes, CEP2- and CEP5 expressing cells showed reduced F-actin localization at the adherens junctions with an increase in thin stress fibers that extended the length of the cell body. Keratinocytes expressing CEPs also showed an altered vinculin distribution and a loss of E-cadherin from adherens junctions. Similar effects were observed in keratinocytes expressing constitutively active Cdc42, but were not seen with a Cdc42/Rac interactive binding domain mutant of CEP2. These results suggest that CEPs act downstream of Cdc42 to induce actin filament assembly leading to cell shape changes. PMID- 11035018 TI - Adrenodoxin reductase homolog (Arh1p) of yeast mitochondria required for iron homeostasis. AB - Arh1p is an essential mitochondrial protein of yeast with reductase activity. Here we show that this protein is involved in iron metabolism. A yeast strain was constructed in which the open reading frame was placed under the control of a galactose-regulated promoter. Protein expression was induced by galactose and repressed to undetectable levels in the absence of galactose, although cells grew quite well in the absence of inducer. Under noninducing conditions, cellular iron uptake was dysregulated, exhibiting a failure to repress in response to medium iron. Iron trafficking within the cell was also disturbed. Exposure of Arh1p depleted cells to increasing iron concentrations during growth led to drastic increases in mitochondrial iron, indicating a loss of homeostatic control. Activity of aconitase, a prototype Fe-S protein, was deficient at all concentrations of mitochondrial iron, although the protein level was unaltered. Heme protein deficiencies were exacerbated in the iron-loaded mitochondria, suggesting a toxic side effect of accumulated iron. Finally, a time course correlated the cellular depletion of Arh1p with the coordinated appearance of various mutant phenotypes including dysregulated cellular iron uptake, deficiency of Fe-S protein activities in mitochondria and cytoplasm, and deficiency of hemoproteins. Thus, Arh1p is required for control of cellular and mitochondrial iron levels and for the activities of Fe-S cluster proteins. PMID- 11035019 TI - DNA recognition by the methyl-CpG binding domain of MeCP2. AB - The methyl-CpG binding domain (MBD) of the transcriptional repressor MeCP2 has been proposed to recognize a single symmetrically methylated CpG base pair via hydrophobic patches on an otherwise positively charged DNA binding surface. We have tested this binding model by analysis of mutant derivatives of the MeCP2 MBD in electrophoretic mobility shift assays complemented by NMR structural analysis. Exposed arginine side chains on the binding face, in particular Arg-111, were found to be critical for binding. Arg-111 was found to interact with the conserved aspartate side chain Asp-121, which is proposed to orientate the arginine side chain to allow specific contacts with the DNA. The conformational flexibility of the disordered B-C loop region, which forms part of the binding face, was also shown to be important. In contrast, mutation of the exposed hydrophobic side chains had a less severe effect on DNA binding. This suggests that the Arg-111 side chain may contribute to sequence-specific recognition of the CpG site rather than simply making nonspecific contacts with the phosphate backbone. The majority of missense mutations within the MBD found in the human genetic disorder Rett syndrome were shown or predicted to affect folding of the domain rather than the DNA recognition event directly. PMID- 11035020 TI - Heme ligation and conformational plasticity in the isolated c domain of cytochrome cd1 nitrite reductase. AB - The heme ligation in the isolated c domain of Paracoccus pantotrophus cytochrome cd(1) nitrite reductase has been characterized in both oxidation states in solution by NMR spectroscopy. In the reduced form, the heme ligands are His69 Met106, and the tertiary structure around the c heme is similar to that found in reduced crystals of intact cytochrome cd1 nitrite reductase. In the oxidized state, however, the structure of the isolated c domain is different from the structure seen in oxidized crystals of intact cytochrome cd1, where the c heme ligands are His69-His17. An equilibrium mixture of heme ligands is present in isolated oxidized c domain. Two-dimensional exchange NMR spectroscopy shows that the dominant species has His69-Met106 ligation, similar to reduced c domains. This form is in equilibrium with a high-spin form in which Met106 has left the heme iron. Melting studies show that the midpoint of unfolding of the isolated c domain is 320.9 +/- 1.2 K in the oxidized and 357.7 +/- 0.6 K in the reduced form. The thermally denatured forms are high-spin in both oxidation states. The results reveal how redox changes modulate conformational plasticity around the c heme and show the first key steps in the mechanism that lead to ligand switching in the holoenzyme. This process is not solely a function of the properties of the c domain. The role of the d1 heme in guiding His17 to the c heme in the oxidized holoenzyme is discussed. PMID- 11035021 TI - Glyceraldehyde-3-phosphate dehydrogenase is required for vesicular transport in the early secretory pathway. AB - Protein transport in the early secretory pathway requires Rab2 GTPase. This protein promotes the recruitment of soluble components that participate in protein sorting and recycling from pre-Golgi intermediates (vesicular tubular clusters (VTCs)). We previously reported that a constitutively activated form of Rab2 (Q65L) as well as Rab2 wild type promoted vesicle formation from VTCs. These vesicles contained Rab2, beta-COP, p53/gp58, and protein kinase Ciota/lambda but lacked anterograde-directed cargo. To identify other candidate Rab2 effectors, the polypeptide composition of the vesicles was further analyzed. We found that vesicles released in response to Rab2 also contained the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). To study the relationship of this enzyme to Rab2 function, we performed a quantitative binding assay to measure recruitment of GAPDH to membrane when incubated with Rab2. Rab2-treated microsomes showed a 5-10-fold increase in the level of membrane-associated GAPDH. We generated an affinity-purified anti-GAPDH polyclonal to study the biochemical role of GAPDH in the early secretory pathway. The antibody arrests transport of a reporter molecule in an assay that reconstitutes ER to Golgi traffic. Furthermore, the affinity-purified antibody blocked the ability of Rab2 to recruit GAPDH to membrane. However, the antibody did not interfere with Rab2 stimulated vesicle release. These data suggest that GAPDH is required for ER to Golgi transport. We propose that membranes incubated with anti-GAPDH and Rab2 form "dead end" vesicles that are unable to transport and fuse with the acceptor compartment. PMID- 11035022 TI - Magnesium ion-mediated binding to tRNA by an amino-terminal peptide of a class II tRNA synthetase. AB - Aspartyl-tRNA synthetase is a class II tRNA synthetase and occurs in a multisynthetase complex in mammalian cells. Human Asp-tRNA synthetase contains a short 32-residue amino-terminal extension that can control the release of charged tRNA and its direct transfer to elongation factor 1 alpha; however, whether the extension binds to tRNA directly or interacts with the synthetase active site is not known. Full-length human AspRS, but not amino-terminal 32 residue-deleted, fully active AspRS, was found to bind to noncognate tRNA(fMet) in the presence of Mg(2+). Synthetic amino-terminal peptides bound similarly to tRNA(fMet), whereas little or no binding of polynucleotides, poly(dA-dT), or polyphosphate to the peptides was found. The apparent binding constants to tRNA by the peptide increased with increasing concentrations of Mg(2+), suggesting Mg(2+) mediates the binding as a new mode of RNA.peptide interactions. The binding of tRNA(fMet) to amino-terminal peptides was also observed using fluorescence-labeled tRNAs and circular dichroism. These results suggest that a small peptide can bind to tRNA selectively and that evolution of class II tRNA synthetases may involve structural changes of amino-terminal extensions for enhanced selective binding of tRNA. PMID- 11035024 TI - Residues lining the inner pore vestibule of human muscle chloride channels. AB - Chloride channels belonging to the ClC family are ubiquitous and participate in a wide variety of physiological and pathophysiological processes. To define sequence segments in ClC channels that contribute to the formation of their ion conduction pathway, we employed a combination of site-directed mutagenesis, heterologous expression, patch clamp recordings, and chemical modification of the human muscle ClC isoform, hClC-1. We demonstrate that a highly conserved 8-amino acid motif (P3) located in the linker between transmembrane domains D2 and D3 contributes to the formation of a wide pore vestibule facing the cell interior. Similar to a previously defined pore region (P1 region), this segment functionally interacts with the corresponding segment of the contralateral subunit. The use of cysteine-specific reagents of different size revealed marked differences in the diameter of pore-forming regions implying that ClC channels exhibit a pore architecture quite similar to that of certain cation channels, in which a narrow constriction containing major structural determinants of ion selectivity is neighbored by wide vestibules on both sides of the membrane. PMID- 11035023 TI - Association with the nuclear matrix and interaction with Groucho and RUNX proteins regulate the transcription repression activity of the basic helix loop helix factor Hes1. AB - Hairy/Enhancer of split 1 (Hes1) is a mammalian transcriptional repressor that plays crucial roles in the regulation of several developmental processes, including neuronal differentiation. The aim of this study was to elucidate the molecular mechanisms that regulate the transcription repression activity of Hes1. It is shown here that Hes1 associates with the nuclear matrix, the ribonucleoprotein network of the nucleus that plays important roles in transcriptional regulation. Nuclear matrix binding is mediated by the same Hes1 C terminal domain that is also required for transcriptional repression. This domain contains the WRPW motif that acts as a binding site for the transcriptional corepressor Groucho, which also localizes to the nuclear matrix. Both the nuclear matrix association and transcription repression activity of Hes1 are inhibited by deletion of the WRPW motif, indicating that Groucho acts as a transcriptional corepressor for Hes1. This corepressor role is not modulated by the Groucho related gene product Grg5. In contrast, the Runt-related protein RUNX2, which localizes to the nuclear matrix and interacts with Groucho and Hes1, can inhibit both the Groucho.Hes1 interaction and the transcription repression ability of Hes1. Together, these observations suggest that transcriptional repression by Hes1 requires interactions with Groucho at the nuclear matrix and that RUNX proteins act as negative regulators of the repressive activity of Groucho.Hes1 complexes. PMID- 11035025 TI - Hormone binding by protein disulfide isomerase, a high capacity hormone reservoir of the endoplasmic reticulum. AB - Protein disulfide isomerase (PDI) is a folding assistant of the eukaryotic endoplasmic reticulum, but it also binds the hormones, estradiol, and 3,3',5 triiodo-l-thyronine (T(3)). Hormone binding could be at discrete hormone binding sites, or it could be a nonphysiological consequence of binding site(s) that are involved in the interaction PDI with its peptide and protein substrates. Equilibrium dialysis, fluorescent hydrophobic probe binding (4,4'-dianilino-1,1' binaphthyl-5,5'-disulfonic acid (bis-ANS)), competition binding, and enzyme activity assays reveal that the hormone binding sites are distinct from the peptide/protein binding sites. PDI has one estradiol binding site with modest affinity (2.1 +/- 0.5 microm). There are two binding sites with comparable affinity for T(3) (4.3 +/- 1.4 microm). One of these overlaps the estradiol site, whereas the other binds the hydrophobic probe, bis-ANS. Neither estradiol nor T(3) inhibit the catalytic or chaperone activity of PDI. Although the affinity of PDI for the hormones estradiol and T(3) is modest, the high local concentration of PDI in the endoplasmic reticulum (>200 microm) would drive hormone binding and result in the association of a substantial fraction (>90%) of the hormones in the cell with PDI. High capacity, low affinity hormone sites may function to buffer hormone concentration in the cell and allow tight, specific binding to the true receptor while preserving a reasonable number of hormone molecules in the very small volume of the cellular environment. PMID- 11035026 TI - Subunit structure of a mammalian ER/Golgi SNARE complex. AB - SNAP receptor (SNARE) complexes bridge opposing membranes to promote membrane fusion within the secretory and endosomal pathways. Because only the exocytic SNARE complexes have been characterized in detail, the structural features shared by SNARE complexes from different fusion steps are not known. We now describe the subunit structure, assembly, and regulation of a quaternary SNARE complex, which appears to mediate an early step in endoplasmic reticulum (ER) to Golgi transport. Purified recombinant syntaxin 5, membrin, and rbet1, three Q-SNAREs, assemble cooperatively to create a high affinity binding site for sec22b, an R SNARE. The syntaxin 5 amino-terminal domain potently inhibits SNARE complex assembly. The ER/Golgi quaternary complex is remarkably similar to the synaptic complex, suggesting that a common pattern is followed at all transport steps, where three Q-helices assemble to form a high affinity binding site for a fourth R-helix on an opposing membrane. Interestingly, although sec22b binds to the combination of syntaxin 5, membrin, and rbet1, it can only bind if it is present while the others assemble; sec22b cannot bind to a pre-assembled ternary complex of syntaxin 5, membrin, and rbet1. Finally, we demonstrate that the quaternary complex containing sec22b is not an in vitro entity only, but is a bona fide species in living cells. PMID- 11035027 TI - 2'-O,4'-C-methylene bridged nucleic acid modification promotes pyrimidine motif triplex DNA formation at physiological pH: thermodynamic and kinetic studies. AB - Extreme instability of pyrimidine motif triplex DNA at physiological pH severely limits its use in an artificial control of gene expression in vivo. Stabilization of the pyrimidine motif triplex at physiological pH is, therefore, crucial in improving its therapeutic potential. To this end, we have investigated the thermodynamic and kinetic effects of our previously reported chemical modification, 2'-O,4'-C-methylene bridged nucleic acid (2',4'-BNA) modification of triplex-forming oligonucleotide (TFO), on pyrimidine motif triplex formation at physiological pH. The thermodynamic analyses indicated that the 2',4'-BNA modification of TFO increased the binding constant of the pyrimidine motif triplex formation at neutral pH by approximately 20 times. The number and position of the 2',4'-BNA modification introduced into the TFO did not significantly affect the magnitude of the increase in the binding constant. The consideration of the observed thermodynamic parameters suggested that the increased rigidity itself of the 2',4'-BNA-modified TFO in the free state relative to the unmodified TFO may enable the significant increase in the binding constant at neutral pH. Kinetic data demonstrated that the observed increase in the binding constant at neutral pH by the 2',4'-BNA modification of TFO resulted from the considerable decrease in the dissociation rate constant. Our results certainly support the idea that the 2',4'-BNA modification of TFO could be a key chemical modification and may eventually lead to progress in therapeutic applications of the antigene strategy in vivo. PMID- 11035028 TI - Identification of distinct signaling pathways leading to the phosphorylation of interferon regulatory factor 3. AB - Infection of host cells by viruses leads to the activation of multiple signaling pathways, resulting in the expression of host genes involved in the establishment of the antiviral state. Among the transcription factors mediating the immediate response to virus is interferon regulatory factor-3 (IRF-3) which is post translationally modified as a result of virus infection. Phosphorylation of latent cytoplasmic IRF-3 on serine and threonine residues in the C-terminal region leads to dimerization, cytoplasmic to nuclear translocation, association with the p300/CBP coactivator, and stimulation of DNA binding and transcriptional activities. We now demonstrate that IRF-3 is a phosphoprotein that is uniquely activated via virus-dependent C-terminal phosphorylation. Paramyxoviridae including measles virus and rhabdoviridae, vesicular stomatitis virus, are potent inducers of a unique virus-activated kinase activity. In contrast, stress inducers, growth factors, DNA-damaging agents, and cytokines do not induce C terminal IRF-3 phosphorylation, translocation or transactivation, but rather activate a MAPKKK-related signaling pathway that results in N-terminal IRF-3 phosphorylation. The failure of numerous well characterized pharmacological inhibitors to abrogate virus-induced IRF-3 phosphorylation suggests the involvement of a novel kinase activity in IRF-3 regulation by viruses. PMID- 11035029 TI - Identification of the functional interleukin-22 (IL-22) receptor complex: the IL 10R2 chain (IL-10Rbeta ) is a common chain of both the IL-10 and IL-22 (IL-10 related T cell-derived inducible factor, IL-TIF) receptor complexes. AB - Interleukin-10 (IL-10)-related T cell-derived inducible factor (IL-TIF; provisionally designated IL-22) is a cytokine with limited homology to IL-10. We report here the identification of a functional IL-TIF receptor complex that consists of two receptor chains, the orphan CRF2-9 and IL-10R2, the second chain of the IL-10 receptor complex. Expression of the CRF2-9 chain in monkey COS cells renders them sensitive to IL-TIF. However, in hamster cells both chains, CRF2-9 and IL-10R2, must be expressed to assemble the functional IL-TIF receptor complex. The CRF2-9 chain (or the IL-TIF-R1 chain) is responsible for Stat recruitment. Substitution of the CRF2-9 intracellular domain with the IFN-gammaR1 intracellular domain changes the pattern of IL-TIF-induced Stat activation. The CRF2-9 gene is expressed in normal liver and kidney, suggesting a possible role for IL-TIF in regulating gene expression in these tissues. Each chain, CRF2-9 and IL-10R2, is capable of binding IL-TIF independently and can be cross-linked to the radiolabeled IL-TIF. However, binding of IL-TIF to the receptor complex is greater than binding to either receptor chain alone. Sharing of the common IL 10R2 chain between the IL-10 and IL-TIF receptor complexes is the first such case for receptor complexes with chains belonging to the class II cytokine receptor family, establishing a novel paradigm for IL-10-related ligands similar to the shared use of the gamma common chain (gamma(c)) by several cytokines, including IL-2, IL-4, IL-7, IL-9, and IL-15. PMID- 11035030 TI - Gonadotropin-releasing hormone receptor microaggregation. Rate monitored by fluorescence resonance energy transfer. AB - Gonadotropin-releasing hormone (GnRH) regulates pituitary gonadotropin release and is a therapeutic target for human and animal reproductive diseases. In the present study we have utilized the technique of fluorescence resonance energy transfer to monitor the rate of GnRH receptor-receptor interactions. This technique relies on the observation that the degree of physical intimacy of molecules can be assessed by the tendency of proximal fluorophores to exchange energy. Our data indicate that GnRH agonist, but not antagonist, occupancy of the GnRH receptor promotes physical intimacy (microaggregation) between receptors. The time course indicates that this occurs promptly (<1 min) after occupancy and persists for at least 80 min and within the physiologically relevant range of the releasing hormone. The process measured is not inhibited by 0.1 mm vinblastin, 2 microm cytochalasin D, or 3 mm EGTA, an observation that distinguishes it from macroaggregation (patching, capping, and internalization). These observations, along with reports from other laboratories, are consonant with a growing body of evidence that indicates that microaggregation is an early event following agonist occupancy of the receptor and part of the mechanism by which effector regulation occurs. PMID- 11035031 TI - The glutathione transferase structural family includes a nuclear chloride channel and a ryanodine receptor calcium release channel modulator. AB - The ubiquitous glutathione transferases (GSTs) catalyze glutathione conjugation to many compounds and have other diverse functions that continue to be discovered. We noticed sequence similarities between Omega class GSTs and a nuclear chloride channel, NCC27 (CLIC1), and show here that NCC27 belongs to the GST structural family. The structural homology prompted us to investigate whether the human Omega class glutathione transferase GSTO1-1 forms or modulates ion channels. We find that GSTO1-1 modulates ryanodine receptors (RyR), which are calcium channels in the endoplasmic reticulum of various cells. Cardiac RyR2 activity was inhibited by GSTO1-1, whereas skeletal muscle RyR1 activity was potentiated. An enzymatically active conformation of GSTO1-1 was required for inhibition of RyR2, and mutation of the active site cysteine (Cys-32 --> Ala) abolished the inhibitory activity. We propose a novel role for GSTO1-1 in protecting cells containing RyR2 from apoptosis induced by Ca(2+) mobilization from intracellular stores. PMID- 11035032 TI - Transcriptional regulation of the yeast gmp synthesis pathway by its end products. AB - AMP and GMP are synthesized from IMP by specific conserved pathways. In yeast, whereas IMP and AMP synthesis are coregulated, we found that the GMP synthesis pathway is specifically regulated. Transcription of the IMD genes, encoding the yeast homologs of IMP dehydrogenase, was repressed by extracellular guanine. Only this first step of GDP synthesis pathway is regulated, since the latter steps, encoded by the GUA1 and GUK1 genes, are guanine-insensitive. Use of mutants affecting GDP metabolism revealed that guanine had to be transformed into GDP to allow repression of the IMD genes. IMD gene transcription was also strongly activated by mycophenolic acid (MPA), a specific inhibitor of IMP dehydrogenase activity. Serial deletions of the IMD2 gene promoter revealed the presence of a negative cis-element, required for guanine regulation. Point mutations in this guanine response element strongly enhanced IMD2 expression, also making it insensitive to guanine and MPA. From these data, we propose that the guanine response element sequence mediates a repression process, which is enhanced by guanine addition, through GDP or a GDP derivative, and abolished in the presence of MPA. PMID- 11035033 TI - The p115-interactive proteins GM130 and giantin participate in endoplasmic reticulum-Golgi traffic. AB - The transport factor p115 is essential for endoplasmic reticulum (ER) to Golgi traffic. P115 interacts with two Golgi proteins, GM130 and giantin, suggesting that they might also participate in ER-Golgi traffic. Here, we show that peptides containing the GM130 or the giantin p115 binding domain and anti-GM130 and anti giantin antibodies inhibit transport of vesicular stomatitis virus (VSV)-G protein to a mannosidase II-containing Golgi compartment. To determine whether p115, GM130, and giantin act together or sequentially during transport, we compared kinetics of traffic inhibition. Anti-p115, anti-GM130, and anti-giantin antibodies inhibited transport at temporally distinct steps, with the p115 requiring step before the GM130-requiring stage, and both preceding the giantin requiring stage. Examination of the distribution of the arrested VSV-G protein showed that anti-p115 antibodies inhibited transport at the level of vesicular tubular clusters, whereas anti-GM130 and anti-giantin antibodies inhibited after the VSV-G protein moved to the Golgi complex. Our results provide the first evidence that GM130 and giantin are required for the delivery of a cargo protein to the mannosidase II-containing Golgi compartment. These data are most consistent with a model where transport from the ER to the cis/medial-Golgi compartments requires the action of p115, GM130, and giantin in a sequential rather than coordinate mechanism. PMID- 11035034 TI - Isolation of a 40-kDa Huntingtin-associated protein. AB - Huntington's disease is caused by an expanded CAG trinucleotide repeat coding for a polyglutamine stretch within the huntingtin protein. Currently, the function of normal huntingtin and the mechanism by which expanded huntingtin causes selective neurotoxicity remain unknown. Clues may come from the identification of huntingtin-associated proteins (HAPs). Here, we show that huntingtin copurifies with a single novel 40-kDa protein termed HAP40. HAP40 is encoded by the open reading frame factor VIII-associated gene A (F8A) located within intron 22 of the factor VIII gene. In transfected cell extracts, HAP40 coimmunoprecipitates with full-length huntingtin but not with an N-terminal huntingtin fragment. Recombinant HAP40 is cytoplasmic in the presence of huntingtin but is actively targeted to the nucleus in the absence of huntingtin. These data indicate that HAP40 is likely to contribute to the function of normal huntingtin and is a candidate for involvement in the aberrant nuclear localization of mutant huntingtin found in degenerating neurons in Huntington's disease. PMID- 11035035 TI - Glutaredoxin protects cerebellar granule neurons from dopamine-induced apoptosis by activating NF-kappa B via Ref-1. AB - The neurotransmitter dopamine (DA) induces apoptosis via its oxidative metabolites. This study shows that glutaredoxin 2 (Grx2) from Escherichia coli and human glutaredoxin could protect cerebellar granule neurons from DA-induced apoptosis. E. coli Grx2, which catalyzes glutathione-disulfide oxidoreduction via its -Cys-Pro-Tyr-Cys- active site, penetrates into cerebellar granule neurons and exerts its activity via NF-kappaB activation. Analysis of single and double cysteine to serine substitutions in the active site of Grx2 showed that both cysteine residues were essential for activity. Although DA significantly reduced NF-kappaB binding activity, Grx2 could stimulate the binding of NF-kappaB to DNA by: (i) translocating NF-kappaB from the cytoplasm to the nucleus after promoting the phosphorylation and degradation of I-kappaBalpha, and (ii) activating the binding of pre existing nuclear NF-kappaB. The DNA binding activity of NF-kappaB itself was essential for neuronal survival. Overexpression of I-kappaB dominant negative gene (I-kappaB-DeltaN) in granule neurons significantly reduced their viability, irrespective of the presence of Grx2. Ref-1 expression was down regulated by DA but up-regulated by Grx2, while treatment of neurons with Ref-1 antisense oligonucleotide reduced the ability of Grx2 to activate NF-kappaB binding activity. These results show that Grx2 exerts its anti apoptotic activity through the activation of Ref-1, which then activates NF-kappaB. PMID- 11035036 TI - Fidelity of uracil-initiated base excision DNA repair in Escherichia coli cell extracts. AB - The error frequency and mutational specificity associated with Escherichia coli uracil-initiated base excision repair were measured using an M13mp2 lacZalpha DNA based reversion assay. Repair was detected in cell-free extracts utilizing a form I DNA substrate containing a site-specific uracil residue. The rate and extent of complete uracil-DNA repair were measured using uracil-DNA glycosylase (Ung)- or double-strand uracil-DNA glycosylase (Dug)-proficient and -deficient isogenic E. coli cells. In reactions utilizing E. coli NR8051 (ung(+) dug(+)), approximately 80% of the uracil-DNA was repaired, whereas about 20% repair was observed using NR8052 (ung(-) dug(+)) cells. The Ung-deficient reaction was insensitive to inhibition by the PBS2 uracil-DNA glycosylase inhibitor protein, implying the involvement of Dug activity. Under both conditions, repaired form I DNA accumulated in conjunction with limited DNA synthesis associated with a repair patch size of 1-20 nucleotides. Reactions conducted with E. coli BH156 (ung(-) dug(+)), BH157 (ung(+) dug(-)), and BH158 (ung(-) dug(-)) cells provided direct evidence for the involvement of Dug in uracil-DNA repair. The rate of repair was 5-fold greater in the Ung-proficient than in the Ung-deficient reactions, while repair was not detected in reactions deficient in both Ung and Dug. The base substitution reversion frequency associated with uracil-DNA repair was determined to be approximately 5.5 x 10(-)(4) with transversion mutations dominating the mutational spectrum. In the presence of Dug, inactivation of Ung resulted in up to a 7.3-fold increase in mutation frequency without a dramatic change in mutational specificity. PMID- 11035037 TI - Mutations in the TATA-binding protein, affecting transcriptional activation, show synthetic lethality with the TAF145 gene lacking the TAF N-terminal domain in Saccharomyces cerevisiae. AB - The general transcription factor TFIID, which is composed of the TATA box-binding protein (TBP) and a set of TBP-associated factors (TAFs), is crucial for both basal and regulated transcription by RNA polymerase II. The N-terminal small segment of yeast TAF145 (yTAF145) binds to TBP and thereby inhibits TBP function. To understand the physiological role of this inhibitory domain, which is designated as TAND (TAF N-terminal domain), we screened mutations, synthetically lethal with the TAF145 gene lacking TAND (taf145 Delta TAND), in Saccharomyces cerevisiae by exploiting a red/white colony-sectoring assay. Our screen yielded several recessive nsl (Delta TAND synthetic lethal) mutations, two of which, nsl1 1 and nsl1-2, define the same complementation group. The NSL1 gene was found to be identical to the SPT15 gene encoding TBP. Interestingly, both temperature sensitive nsl1/spt15 alleles, which harbor the single amino acid substitutions, S118L and P65S, respectively, were defective in transcriptional activation in vivo. Several other previously characterized activation-deficient spt15 alleles also displayed synthetic lethal interactions with taf145 Delta TAND, indicating that TAND and TBP carry an overlapping but as yet unidentified function that is specifically required for transcriptional regulation. PMID- 11035038 TI - Neonatal mortality in an aquaporin-2 knock-in mouse model of recessive nephrogenic diabetes insipidus. AB - Hereditary non-X-linked nephrogenic diabetes insipidus (NDI) is caused by mutations in the aquaporin-2 (AQP2) water channel. In transfected cells, the human disease-causing mutant AQP2-T126M is retained at the endoplasmic reticulum (ER) where it is functional and targetable to the plasma membrane with chemical chaperones. A mouse knock-in model of NDI was generated by targeted gene replacement using a Cre-loxP strategy. Along with T126M, mutations H122S, N124S, and A125T were introduced to preserve the consensus sequence for N-linked glycosylation found in human AQP2. Breeding of heterozygous mice yielded the expected Mendelian distribution with 26 homozygous mutant offspring of 99 live births. The mutant mice appeared normal at 2-3 days after birth but failed to thrive and generally died by day 6 if not given supplemental fluid. Urine/serum analysis showed a urinary concentrating defect with serum hyperosmolality and low urine osmolality that was not increased by a V2 vasopressin agonist. Northern blot analysis showed up-regulated AQP2-T126M transcripts of identical size to wild-type AQP2. Immunoblots showed complex glycosylation of wild-type AQP2 but mainly endoglycosidase H-sensitive core glycosylation of AQP2-T126M indicating ER retention. Biochemical analysis revealed that the AQP2-T126M protein was resistant to detergent solubilization. Kidneys from mutant mice showed collecting duct dilatation, papillary atrophy, and unexpectedly, some plasma membrane AQP2 staining. The severe phenotype of the AQP2 mutant mice compared with that of mice lacking kidney water channels AQP1, AQP3, and AQP4 indicates a critical role for AQP2 in neonatal renal function in mice. Our results establish a mouse model of human autosomal NDI and provide the first in vivo biochemical data on a disease causing AQP2 mutant. PMID- 11035039 TI - The ectodermal dysplasia receptor activates the nuclear factor-kappaB, JNK, and cell death pathways and binds to ectodysplasin A. AB - The ectodermal dysplasia receptor (EDAR) is a recently isolated member of the tumor necrosis factor receptor family that has been shown to play a key role in the process of ectodermal differentiation. We present evidence that EDAR is capable of activating the nuclear factor-kappaB, JNK, and caspase-independent cell death pathways and that these activities are impaired in mutants lacking its death domain or those associated with anhidrotic ectodermal dysplasia and the downless phenotype. Although EDAR possesses a death domain, it did not interact with the death domain-containing adaptor proteins TRADD and FADD. EDAR successfully interacted with various TRAF family members; however, a dominant negative mutant of TRAF2 was incapable of blocking EDAR-induced nuclear factor kappaB or JNK activation. Collectively, the above results suggest that EDAR utilizes a novel signal transduction pathway. Finally, ectodysplasin A can physically interact with the extracellular domain of EDAR and thus represents its biological ligand. PMID- 11035040 TI - Altered processing of fibronectin in mice lacking heparin. a role for heparin dependent mast cell chymase in fibronectin degradation. AB - We have previously generated a mouse strain with a defect in its heparin biosynthesis by targeting the gene for N-deacetylase/N-sulfotransferase-2 (NDST 2). The NDST-2(-/-) mice show reduced levels of various mast cell mediators such as histamine and various heparin-binding mast cell proteases, including chymases, tryptases, and carboxypeptidase A. In this work we have addressed the possible functional consequences of the lack of sulfated heparin. Peritoneal cells were harvested from normal and NDST-2(-/-) mice. After culturing the cells, conditioned media were collected and were subjected to SDS-polyacrylamide gel electrophoresis under reducing conditions. Several differences in the protein patterns were observed, including the presence of large amounts of a approximately 250-kDa protein in medium from NDST-2(-/-) mice that was absent in normal controls. Peptide microsequencing revealed identity of this protein with fibronectin. Western blot analysis showed the presence of fibronectin degradation products in cell cultures from normal mice, which were absent in cultures from NDST-2(-/-) animals. Further experiments showed that the degradation of fibronectin observed in cell cultures from NDST-2(+/+) mice was catalyzed by mast cell chymase in a strongly heparin-dependent manner. This report thus indicates a biological function for chymase/heparin proteoglycan complexes in fibronectin turnover. PMID- 11035041 TI - Ionizing radiation down-regulates p53 protein in primary Egr-1-/- mouse embryonic fibroblast cells causing enhanced resistance to apoptosis. AB - In this study, we sought to investigate the mechanism of the proapoptotic function of Egr-1 in relation to p53 status in normal isogenic cell backgrounds by using primary MEF cells established from homozygous (Egr-1(-/-)) and heterozygous (Egr-1(+/-)) Egr-1 knock-out mice. Ionizing radiation caused significantly enhanced apoptosis in Egr-1(+/-) cells (22.8%; p < 0.0001) when compared with Egr-1(-/-) cells (3.5%). Radiation elevated p53 protein in Egr-1(+/ ) cells in 3-6 h. However, in Egr-1(-/-) cells, the p53 protein was down regulated 1 h after radiation and was completely degraded at the later time points. Radiation elevated the p53-CAT activity in Egr-1(+/-) cells but not in Egr-1(-/-) cells. Interestingly, transient overexpression of EGR-1 in p53(-/-) MEF cells caused marginal induction of radiation-induced apoptosis when compared with p53(+/+) MEF cells. Together, these results indicate that Egr-1 may transregulate p53, and both EGR-1 and p53 functions are essential to mediate radiation-induced apoptosis. Rb, an Egr-1 target gene, forms a trimeric complex with p53 and MDM2 to prevent MDM2-mediated p53 degradation. Low levels of Rb including hypophosphorylated forms were observed in Egr-1(-/-) MEF cells before and after radiation when compared with the levels observed in Egr-1(+/-) cells. Elevated amounts of the p53-MDM2 complex and low amounts of Rb-MDM-2 complex were observed in Egr-1(-/-) cells after radiation. Because of a reduction in Rb binding to MDM2 and an increase in MDM2 binding with p53, p53 is directly degraded by MDM2, and this leads to inactivation of the p53-mediated apoptotic pathway in Egr-1(-/-) MEF cells. Thus, the proapoptotic function of Egr-1 may involve the mediation of Rb protein that is essential to overcome the antiapoptotic function of MDM2 on p53. PMID- 11035042 TI - Erythrocyte water permeability and renal function in double knockout mice lacking aquaporin-1 and aquaporin-3. AB - Aquaporin (AQP) water channel AQP3 has been proposed to be the major glycerol and non-AQP1 water transporter in erythrocytes. AQP1 and AQP3 are also expressed in the kidney where their deletion in mice produces distinct forms of nephrogenic diabetes insipidus. Here AQP1/AQP3 double knockout mice were generated and analyzed to investigate the functional role of AQP3 in erythrocytes and kidneys. 53 double knockout mice were born out of 756 pups from breeding double heterozygous mice. The double knockout mice had reduced survival and impaired growth compared with the single knockout mice. Erythrocyte water permeability was 7-fold reduced by AQP1 deletion but not further reduced in AQP1/AQP3 null mice. AQP3 deletion did not affect erythrocyte glycerol permeability or its inhibition by phloretin. Daily urine output in AQP1/AQP3 double knockout mice (15 ml) was 9 fold greater than in wild-type mice, and urine osmolality (194 mosm) was 8.4-fold reduced. The mice remained polyuric after DDAVP administration or water deprivation. The renal medulla in most AQP1/AQP3 null mice by age 4 weeks was atrophic and fluid-filled due to the severe polyuria and hydronephrosis. Our data provide direct evidence that AQP3 is not functionally important in erythrocyte water or glycerol permeability. The renal function studies indicate independent roles of AQP1 and AQP3 in countercurrent exchange and collecting duct osmotic equilibration, respectively. PMID- 11035043 TI - Stable SNARE complex prior to evoked synaptic vesicle fusion revealed by fluorescence resonance energy transfer. AB - Although it is clear that soluble N-ethylmaleimide-sensitive factor (NSF) attachment protein receptor (SNARE) complex plays an essential role in synaptic vesicle fusion, the dynamics of SNARE assembly during vesicle fusion remain to be determined. In this report, we employ fluorescence resonance energy transfer technique to study the formation of SNARE complexes. Donor/acceptor pair variants of green fluorescent protein (GFP), cyan fluorescent protein (CFP), and yellow fluorescent protein (YFP) are fused with the N termini of SNAP-25 and synaptobrevin, respectively. In vitro assembly of SNARE core complex in the presence of syntaxin shows strong fluorescence resonance energy transfer (FRET) between the CFP-SNAP-25 and YFP-synaptobrevin. Under the same conditions, CFP fused to the C terminus of SNAP-25, and YFP- synaptobrevin have no FRET. Adenovirus-mediated gene transfer is used to express the fusion proteins in PC12 cells and cultured rat cerebellar granule cells. Strong FRET is associated with neurite membranes and vesicular structures in PC12 cells co-expressing CFP-SNAP 25 and YFP-synaptobrevin. In cultured rat cerebellar granule cells, FRET between CFP-SNAP-25 and YFP-synaptobrevin is mostly associated with sites presumed to be synaptic junctions. Neurosecretion in PC12 cells initiated by KCl depolarization leads to an increase in the extent of FRET. These results demonstrate that significant amounts of stable SNARE complex exist prior to evoked synaptic vesicle fusion and that the assembly of SNARE complex occurs during vesicle docking/priming stage. Moreover, it demonstrates that FRET can be used as an effective tool for investigating dynamic SNARE interactions during synaptic vesicle fusion. PMID- 11035044 TI - Calcium binding to calmodulin leads to an N-terminal shift in its binding site on the ryanodine Receptor. AB - The skeletal muscle calcium release channel, ryanodine receptor, is activated by calcium-free calmodulin and inhibited by calcium-bound calmodulin. Previous biochemical studies from our laboratory have shown that calcium-free calmodulin and calcium bound calmodulin protect sites at amino acids 3630 and 3637 from trypsin cleavage (Moore, C. P., Rodney, G., Zhang, J. Z., Santacruz-Toloza, L., Strasburg, G., and Hamilton, S. L. (1999) Biochemistry 38, 8532-8537). We now demonstrate that both calcium-free calmodulin and calcium-bound calmodulin bind with nanomolar affinity to a synthetic peptide matching amino acids 3614-3643 of the ryanodine receptor. Deletion of the last nine amino acids (3635-3643) destroys the ability of the peptide to bind calcium-free calmodulin, but not calcium-bound calmodulin. We propose a novel mechanism for calmodulin's interaction with a target protein. Our data suggest that the binding sites for calcium-free calmodulin and calcium-bound calmodulin are overlapping and, when calcium binds to calmodulin, the calmodulin molecule shifts to a more N-terminal location on the ryanodine receptor converting it from an activator to an inhibitor of the channel. This region of the ryanodine receptor has previously been identified as a site of intersubunit contact, suggesting the possibility that calmodulin regulates ryanodine receptor activity by regulating subunit subunit interactions. PMID- 11035045 TI - The tumor suppressor PTEN is phosphorylated by the protein kinase CK2 at its C terminus. Implications for PTEN stability to proteasome-mediated degradation. AB - The tumor suppressor phosphatase PTEN regulates cell migration, growth, and survival by dephosphorylating phosphatidylinositol second messengers and signaling phosphoproteins. PTEN possesses a C-terminal noncatalytic regulatory domain that contains multiple putative phosphorylation sites, which could play an important role in the control of its biological activity. The protein kinase CK2 phosphorylated, in a constitutive manner, a cluster of Ser/Thr residues located at the PTEN C terminus. PTEN-phosphorylated defective mutants showed decreased stability in comparison with wild type PTEN and were more rapidly degraded by the proteasome. Inhibition of PTEN phosphorylation by the CK2 inhibitor 5,6-dichloro 1-beta-d-ribofuranosyl-benzimidazole also diminished the PTEN protein content. Our results support the notion that proper phosphorylation of PTEN by CK2 is important for PTEN protein stability to proteasome-mediated degradation. PMID- 11035046 TI - Cutting edge: B cell linker protein is dispensable for the allelic exclusion of immunoglobulin heavy chain locus but required for the persistence of CD5+ B cells. AB - The pre-B cell receptor (pre-BCR) and the BCR are required for B lymphopoiesis and for the allelic exclusion of Ig genes. Mice lacking B cell linker (BLNK) protein that is a component of the BCR signaling pathway have impaired B cell development. In this report, we show that allelic exclusion is intact in BLNK(-/ ) mice harboring a V(H)12 transgene. This differs from mice lacking the tyrosine kinase Syk that is upstream of BLNK in BCR signaling and contrasts with mice lacking SLP-76 that is the equivalent adaptor molecule in TCR-signal transduction. We also show that, whereas most wild-type V(H)12-expressing B cells are CD5(+), the majority of the splenic V(H)12-expressing BLNK(-/-) B cells are CD5(-). A small population of V(H)12-expressing, BLNK(-/-) CD5(+) B cells is detectable in the peritoneal cavity of younger but not older mice. This suggests that BLNK deficiency affects not only the generation but also the persistence of B-1 cells. PMID- 11035047 TI - Cutting edge: gab2 mediates an inhibitory phosphatidylinositol 3'-kinase pathway in T cell antigen receptor signaling. AB - Phosphatidylinositol 3'-kinase (PI3K) is a key component of multiple signaling pathways, where it typically promotes survival, proliferation, and/or adhesion. Here, we show that in TCR signaling, the scaffolding adapter Gab2 delivers an inhibitory signal via PI3K. Overexpression of Gab2 in T cell lines inhibits TCR evoked activation of the IL-2 promoter, blocking NF-AT- and NF-kappaB-directed transcription. Inhibition is abrogated by mutating the Gab2 p85-binding sites, by treatment with PI3K inhibitors or by cotransfection of phosphatase homolog of tensin. Our findings provide the first evidence of a negative function for a scaffolding adapter in T cells and identify Gab2/PI3K-containing complexes as novel regulators of TCR signaling. PMID- 11035048 TI - Phosphorothioate backbone modification modulates macrophage activation by CpG DNA. AB - Macrophages respond to unmethylated CpG motifs present in nonmammalian DNA. Stabilized phosphorothioate-modified oligodeoxynucleotides (PS-ODN) containing CpG motifs form the basis of immunotherapeutic agents. In this study, we show that PS-ODN do not perfectly mimic native DNA in activation of macrophages. CpG containing PS-ODN were active at 10- to 100-fold lower concentrations than corresponding phosphodiester ODN in maintenance of cell viability in the absence of CSF-1, in induction of NO production, and in activation of the IL-12 promoter. These enhancing effects are attributable to both increased stability and rate of uptake of the PS-ODN. By contrast, PS-ODN were almost inactive in down-modulation of the CSF-1R from primary macrophages and activation of the HIV-1 LTR. Delayed or poor activation of signaling components may contribute to this, as PS-ODN were slower and less effective at inducing phosphorylation of the extracellular signal related kinases 1 and 2. In addition, at high concentrations, non-CpG PS-ODN specifically inhibited responses to CpG DNA, whereas nonstimulatory phosphodiester ODN had no such effect. Although nonstimulatory PS-ODN caused some inhibition of ODN uptake, this did not adequately explain the levels of inhibition of activity. The results demonstrate that the phosphorothioate backbone has both enhancing and inhibitory effects on macrophage responses to CpG DNA. PMID- 11035050 TI - Activated murine endothelial cells have reduced immunogenicity for CD8+ T cells: a mechanism of immunoregulation? AB - The immunogenic properties of primary cultures of murine lung microvascular endothelial cells (EC) were analyzed. Resting endothelial cells were found to constitutively express low levels of MHC class I and CD80 molecules. IFN-gamma treatment of EC resulted in a marked up-regulation of MHC class I, but no change was observed in the level of CD80 expression. No CD86 molecules were detectable under either condition. The ability of peptide-pulsed EC to induce the proliferation of either the HY-specific, H2-K(k)-restricted CD8(+) T cell clone (C6) or C6 TCR-transgenic naive CD8(+) T cells was analyzed. Resting T cells were stimulated to divide by quiescent peptide-prepulsed EC, while peptide-pulsed, cytokine-activated EC lost the ability to induce T cell division. Furthermore, Ag presentation by cytokine-activated EC induced CD8(+) T cell hyporesponsiveness. The immunogenicity of activated EC could be restored by adding nonsaturating concentrations of anti-H2-K(k) Ab in the presence of an optimal concentration of cognate peptide. This is consistent with the suggestion that the ratio of TCR engagement to costimulation determines the outcome of T cell recognition. In contrast, activated peptide-pulsed EC were killed more efficiently by fully differentiated effector CD8(+) T cells. Finally, evidence is provided that Ag recognition of EC can profoundly affect the transendothelial migration of CD8(+) T cells. Taken together, these results suggest that EC immunogenicity is regulated in a manner that contributes to peripheral tolerance. PMID- 11035049 TI - V gamma 1+ T cells suppress and V gamma 4+ T cells promote susceptibility to coxsackievirus B3-induced myocarditis in mice. AB - Coxsackievirus B3 infections of C57BL/6 mice, which express the MHC class II IA but not IE Ag, results in virus replication in the heart but minimal myocarditis. In contrast, Bl.Tg.Ealpha mice, which are C57BL/6 mice transgenically induced to express IE Ag, develop significant myocarditis upon Coxsackievirus B3 infection. Despite this difference in inflammatory damage, cardiac virus titers are similar between C57BL/6 and Bl.Tg.Ealpha mice. Removing gammadelta T cells from either strain by genetic manipulation (gammadelta knockout(ko)) changes the disease phenotype. C57BL/6 gammadelta ko mice show increased myocarditis. In contrast, Bl.Tg.Ealpha gammadelta ko mice show decreased cardiac inflammation. Flow cytometry revealed a difference in the gammadelta cell subsets in the two strains, with Vgamma1 dominating in C57BL/6 mice, and Vgamma4 predominating Bl.Tg.Ealpha mice. This suggests that these two Vgamma-defined subsets might have different functions. To test this possibility, we used mAb injection to deplete each subset. Mice depleted of Vgamma1 cells showed enhanced myocarditis, whereas those depleted of Vgamma4 cells suppressed myocarditis. Adoptively transfusing enriched Vgamma4(+) cells to the C57BL/6 and Bl.Tg. Ealpha gammadelta ko strains confirmed that the Vgamma4 subset promoted myocarditis. Th subset analysis suggests that Vgamma1(+) cells biased the CD4(+) T cells to a dominant Th2 cell response, whereas Vgamma4(+) cells biased CD4(+) T cells toward a dominant Th1 cell response. PMID- 11035051 TI - Cytokine dysregulation induced by apoptotic cells is a shared characteristic of murine lupus. AB - Of the multiple murine models of autoimmunity, the three most closely resembling human systemic lupus erythematosus (SLE) are the MRL/lpr, New Zealand Black/White F(1), and male BXSB. Although these strains share many disease characteristics, no common cellular defect has previously been found in prediseased mice from all these strains. We show in this study that macrophages from prediseased mice of all three SLE-prone strains, as well as macrophages from mice whose genomes contribute to the development of SLE (MRL/+, New Zealand White, New Zealand Black, female BXSB, and LG/J), have an identical and profound defect in cytokine expression that is triggered by apoptotic cells. Strikingly, none of 13 nonautoimmune strains tested exhibited this defect. Given that apoptotic Ags have been increasingly recognized as the target of autoantibodies, a defect in cytokine expression that is triggered by apoptotic cells has broad potential to upset the balance between tolerance and immunity. PMID- 11035052 TI - A subset of cytolytic dendritic cells in rat. AB - Dendritic cells (DCs) are a rare population of leukocytes specialized in Ag processing and presentation to T cells. We have previously shown that cultured rat splenic DCs exhibit a cytotoxic activity against selected target cells. In this study, we analyzed this function in DCs freshly prepared from lymphoid organs using the DC-specific OX62 mAb and magnetic beads. Freshly extracted splenic DCs, but not lymph node and thymic DCs, exhibited a strong and moderate cytotoxic activity against YAC-1 and K562 target cells, respectively. FACS analyses showed that spleen contained a minor subset (10-15%) of CD4(+) and class II(int) DCs that also expressed the OX41 Ag and the lymphoid-related Ags CD5 and CD90 (Thy-1) and a major (80-85%) subset of CD4(-)/OX41(-)/CD5(-) and class II(int) DCs. The cytotoxic activity of splenic DCs was strictly restricted to the CD4(-) DCs, a subset poorly represented in LN and thymus. Contrasting with our previous report using cultured splenic DCs, freshly isolated splenic DCs killed YAC-1 cells using a Ca(2+)-independent mechanism, but this function did not appear mediated by Fas ligand, TNF-related apoptosis-inducing ligand, or TNF alpha. Therefore, rat DCs contain a subset of naturally cytolytic cells that could play a role in both innate and acquired immune responses. Together with our previous report, these data suggest that rat DCs can use two mechanisms of cytotoxicity depending on their maturation/activation state. PMID- 11035053 TI - A physiological ligand of positive selection is recognized as a weak agonist. AB - Positive selection is a process that ensures that peripheral T cells express TCR that are self-MHC restricted. This process occurs in the thymus and requires both self-MHC and self-peptides. We have recently established a TCR transgenic (TCR(trans)(+)) mouse model using the C10.4 TCR restricted to the MHC class Ib molecule, H2-M3. Having defined H2-M3 as the positively selecting MHC molecule, the severely limited number of H2-M3 binding peptides allowed us to characterize a mitochondrial NADH dehydrogenase subunit 1-derived 9-mer peptide as the physiological ligand of positive selection. Here, we demonstrate that the NADH dehydrogenase subunit 1 self-peptide is seen by mature C10.4 TCR(trans)(+) T cells as a weak agonist and induces positive selection at a defined concentration range. We also found that the full-length cognate peptide, a strong agonist for mature C10.4 TCR(trans)(+) T cells, initiated positive selection, albeit at significantly lower concentrations. At increased peptide concentrations, and thus increased epitope densities, either peptide only induced the development of partially functional T cells. We conclude that successful positive selection only proceeded at a defined, yet fairly narrow window of avidity. PMID- 11035054 TI - Accumulation of RXR alpha during activation of cycling human T lymphocytes: modulation of RXRE transactivation function by mitogen-activated protein kinase pathways. AB - We have previously reported that the activation of resting human immature peripheral blood T (PBT) lymphocytes is associated with the loss of retinoid X receptor alpha (RXRalpha) expression. In the present study, we have demonstrated that, unlike resting cells, activation of cycling human mature PBT lymphocytes, and T lymphocyte leukemia cell lines is accompanied by the accumulation of RXRalpha mRNA and protein. Interestingly, cyclosporin A further augmented RXRalpha expression, indicating the involvement of calcineurin pathways in the process. 9-cis retinoic acid inhibited the accumulation, suggesting that retinoids can regulate the synthesis of their own receptors during T cell activation. Transfection analysis in Jurkat cells, using RXRE-dependent reporter assays, showed that RXRalpha accumulated during T cell activation was transcriptionally inactive. To investigate the mechanism of such inhibition, the role of two mitogen-activated protein kinase pathways, c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK), in modulating RXRE dependent transcription, was explored. The expression of constitutively active MAP/ERK kinase kinase 1 (MEKK1) inhibited RXRE-dependent transcription, whereas dominant negative MEKK1 increased the transcription, indicating the involvement of JNK signaling pathways in the process. In contrast, expression of constitutively active MEK1, which activates ERK pathway, enhanced RXRE-dependent activation. When both were activated simultaneously, JNK pathway was dominant over ERK pathway and resulted in inhibition of RXRE-mediated transcription. These data demonstrate a dual regulatory control of RXRalpha expression during the activation of resting and cycling T lymphocytes and indicate a dynamic balance between JNK and ERK pathways in modulating RXRE-mediated transactivation. PMID- 11035055 TI - The effects of prolonged administration of 5-bromodeoxyuridine on cells of the immune system. AB - We have determined the in vivo effect of 5-bromodeoxyuridine (BrdU) administered to mice in the drinking water for various lengths of time on the performance of T and B lymphocytes in a number of experimental protocols. Young mice continuously exposed to BrdU fail to gain weight, and the lymphocytes recovered after a prolonged period of exposure are fewer in number than in control mice. The recovery of normal levels of T and B lymphocytes after irradiation is severely impaired. Ag-specific cells responding to Ag in an adoptive transfer model fail to expand as much in the presence of BrdU as in the absence, and the Ag-specific effectors produced in the presence of BrdU are less able to secrete cytokines upon restimulation in vitro. Polarized populations of Tc1 and Tc2 effectors generated in vitro proliferate less in the presence of BrdU, and the resulting effectors make less cytokines per cell upon restimulation. Thus, the incorporation of BrdU into T or B lymphocytes can, under some circumstances, seriously impair the performance of the labeled cells, and these findings raise a note of caution in the interpretation of studies that make use of long-term exposure to BrdU. PMID- 11035056 TI - Depletion of CD4 and CD8 T lymphocytes in mice in vivo enhances 1,25 dihydroxyvitamin D3-stimulated osteoclast-like cell formation in vitro by a mechanism that is dependent on prostaglandin synthesis. AB - To investigate the role of T lymphocytes in osteoclastogenesis, we performed in vivo depletion of CD4 and/or CD8 T lymphocyte subsets and evaluated in vitro osteoclast-like cell (OCL) formation. T lymphocyte depletion (TLD) with mAbs was confirmed 24 h later by flow cytometry. OCL formation was stimulated with 1, 25 dihydroxyvitamin D(3) (1,25-(OH)(2)D(3)) in bone marrow and with recombinant mouse (rm) receptor activator of NF-kappaB ligand (RANK-L) and rmM-CSF in bone marrow and spleen cell cultures. OCL formation was up to 2-fold greater in 1,25 (OH)(2)D(3)-stimulated bone marrow cultures from TLD mice than in those from intact mice. In contrast, TLD did not alter OCL formation in bone marrow or spleen cell cultures that were stimulated with rmRANK-L and rmM-CSF. The effects of TLD seemed to be mediated by enhanced PG synthesis, because the PGE(2) concentration in the medium of 1, 25-(OH)(2)D(3)-stimulated bone marrow cultures from TLD mice was 5-fold higher than that in cultures from intact mice, and indomethacin treatment abolished the stimulatory effect of TLD on OCL formation. There was a 2-fold increase in RANK-L expression and an almost complete suppression of osteoprotegerin expression in 1, 25-(OH)(2)D(3)-stimulated bone marrow cultures from TLD mice compared with those from intact mice. Although there was a small (20%) increase in IL-1alpha expression in 1, 25-(OH)(2)D(3) stimulated bone marrow cultures from TLD mice, TLD in mice lacking type I IL-1R and wild-type mice produced similar effects on OCL formation. Our data demonstrate that TLD up-regulates OCL formation in vitro by increasing PG production, which, in turn, produces reciprocal changes in RANK-L and osteoprotegerin expression. These results suggest that T lymphocytes influence osteoclastogenesis by altering bone marrow stromal cell function. PMID- 11035057 TI - Immortalization of human CD8+ T cell clones by ectopic expression of telomerase reverse transcriptase. AB - Replicative senescence of T cells is correlated with erosion of telomere ends. Telomerase plays a key role in maintaining telomere length. Therefore, it is thought that telomerase regulates the life span of T cells. To test this hypothesis, we have over-expressed human telomerase reverse transcriptase in human CD8(+) T cells. Ectopic expression of human telomerase reverse transcriptase led to immortalization of these T cells, without altering the phenotype and without loss of specificity or functionality. As the T cells remained dependent on cytokines and Ag stimulation for their in vitro expansion, we conclude that immortalization was achieved without malignant transformation. PMID- 11035058 TI - Divergent roles for CD4+ T cells in the priming and effector/memory phases of adoptive immunotherapy. AB - The requirement for CD4(+) Th cells in the cross-priming of antitumor CTL is well accepted in tumor immunology. Here we report that the requirement for T cell help can be replaced by local production of GM-CSF at the vaccine site. Experiments using mice in which CD4(+) T cells were eliminated, either by Ab depletion or by gene knockout of the MHC class II beta-chain (MHC II KO), revealed that priming of therapeutic CD8(+) effector T cells following vaccination with a GM-CSF transduced B16BL6-D5 tumor cell line occurred independently of CD4(+) T cell help. The adoptive transfer of CD8(+) effector T cells, but not CD4(+) effector T cells, led to complete regression of pulmonary metastases. Regression of pulmonary metastases did not require either host T cells or NK cells. Transfer of CD8(+) effector T cells alone could cure wild-type animals of systemic tumor; the majority of tumor-bearing mice survived long term after treatment (>100 days). In contrast, adoptive transfer of CD8(+) T cells to tumor-bearing MHC II KO mice improved survival, but eventually all MHC II KO mice succumbed to metastatic disease. WT mice cured by adoptive transfer of CD8(+) T cells were resistant to tumor challenge. Resistance was mediated by CD8(+) T cells in mice at 50 days, while both CD4(+) and CD8(+) T cells were important for protection in mice challenged 150 days following adoptive transfer. Thus, in this tumor model CD4(+) Th cells are not required for the priming phase of CD8(+) effector T cells; however, they are critical for both the complete elimination of tumor and the maintenance of a long term protective antitumor memory response in vivo. PMID- 11035059 TI - Endogenous production of TGF-beta is essential for osteoclastogenesis induced by a combination of receptor activator of NF-kappa B ligand and macrophage-colony stimulating factor. AB - Differentiation of osteoclasts, the cells primarily responsible for bone resorption, is controlled by a variety of osteotropic hormones and cytokines. Of these factors, receptor activator of NF-kappaB (RANK) ligand (RANKL) has been recently cloned as an essential inducer of osteoclastogenesis in the presence of M-CSF. Here, we isolated a stroma-free population of monocyte/macrophage (M/Mphi) like hemopoietic cells from mouse unfractionated bone cells that were capable of differentiating into mature osteoclasts by treatment with soluble RANKL (sRANKL) and M-CSF. However, the efficiency of osteoclast formation was low, suggesting the requirement for additional factors. The isolated M/Mphi-like hemopoietic cells expressed TGF-beta and type I and II receptors of TGF-beta. Therefore, we examined the effect of TGF-beta on osteoclastogenesis. TGF-beta with a combination of sRANKL and M-CSF promoted the differentiation of nearly all M/Mphi like hemopoietic cells into cells of the osteoclast lineage. Neutralizing anti TGF-beta Ab abrogated the osteoclast generation. These TGF-beta effects were also observed in cultures of unfractionated bone cells, and anti-TGF-beta blocked the stimulatory effect of 1, 25-dihydroxyvitamin D(3). Translocation of NF-kappaB into nuclei induced by sRANKL in TGF-beta-pretreated M/Mphi-like hemopoietic cells was greater than that in untreated cells, whereas TGF-beta did not up regulate the expression of RANK, the receptor of RANKL. Our findings suggest that TGF-beta is an essential autocrine factor for osteoclastogenesis. PMID- 11035060 TI - Supra-agonist peptides enhance the reactivation of memory CTL responses. AB - Single amino acid substitutions at TCR contacts may transform a natural peptide Ag in CTL ligands with partial agonist, antagonist, or null activity. We obtained peptide variants by changing nonanchor amino acid residues involved in MHC class I binding. These peptides were derived from a subdominant HLA-A2-presented, latent membrane protein 2-derived epitope expressed in EBV-infected cells and in EBV-associated tumors. We found that small structural changes produced ligands with vastly different activities. In particular, the variants that associated more stably to HLA-A2/molecules did not activate any CTL function, behaving as null ligands. Interestingly, T cell stimulations performed with the combination of null ligands and the natural epitope produced significantly higher specific CTL reactivation than reactivation of CTLs induced by the wild-type epitope alone. In addition, these particular variants activated memory CTL responses in the presence of concentrations of natural epitope that per se did not induce T cell responses. We show here that null ligands increased ZAP-70 tyrosine kinase activation induced by the natural epitope. Our results demonstrate for the first time that particular peptide variants, apparently behaving as null ligands, interact with the TCR, showing a supra-agonist activity. These variant peptides did not affect the effector T cell functions activated by the natural epitope. Supra-agonist peptides represent the counterpart of antagonists and may have important applications in the development of therapeutic peptides. PMID- 11035061 TI - Divergent response to LPS and bacteria in CD14-deficient murine macrophages. AB - Gram-negative bacteria and the LPS constituent of their outer membranes stimulate the release of inflammatory mediators believed to be responsible for the clinical manifestations of septic shock. The GPI-linked membrane protein, CD14, initiates the signaling cascade responsible for the induction of this inflammatory response by LPS. In this paper, we report the generation and characterization of CD14-null mice in which the entire coding region of CD14 was deleted. As expected, LPS failed to elicit TNF-alpha and IL-6 production in macrophages taken from these animals, and this loss in responsiveness is associated with impaired activation of both the NF-kappaB and the c-Jun N-terminal mitogen-activated protein kinase pathways. The binding and uptake of heat-killed Escherichia coli, measured by FACS analysis, did not differ between CD14-null and wild-type macrophages. However, in contrast to the findings with LPS, whole E. coli stimulated similar levels of TNF-alpha release from CD14-null and wild-type macrophages at a dose of 10 bioparticles per cell. This effect was dose dependent, and at lower bacterial concentrations CD14-deficient macrophages produced significantly less TNF-alpha than wild type. Approximately half of this CD14-independent response appeared to be mediated by CD11b/CD18, as demonstrated by receptor blockade using neutrophil inhibitory factor. An inhibitor of phagocytosis, cytochalasin B, abrogated the induction of TNF-alpha in CD14-deficient macrophages by E. coli. These data indicate that CD14 is essential for macrophage responses to free LPS, whereas other receptors, including CD11b/CD18, can compensate for the loss of CD14 in response to whole bacteria. PMID- 11035062 TI - Possible involvement of cyclophilin B and caspase-activated deoxyribonuclease in the induction of chromosomal DNA degradation in TCR-stimulated thymocytes. AB - TCR engagement of immature CD4(+)CD8(+) thymocytes induces clonal maturation (positive selection) as well as clonal deletion (negative selection) in the thymus. However, the cell death execution events of thymocytes during the negative selection process remain obscure. Using a cell-free system, we identified two different DNase activities in the cytosol of in vivo anti-TCR stimulated murine thymocytes: one that induced chromosomal DNA fragmentation, which was inhibited by an inhibitor of caspase-activated DNase, and another that induced plasmid DNA degradation, which was not inhibited by an inhibitor of caspase-activated DNase. We purified the protein to homogeneity that induced plasmid DNA degradation from the cytosol of anti-CD3-stimulated thymocytes and found that it is identical with cyclophilin B (Cyp B), which was reported to locate in endoplasmic reticulum. Ab against Cyp B specifically inhibited the DNA degradation activity in the cytosol of anti-CD3-stimulated thymocytes. Furthermore, recombinant Cyp B induced DNA degradation of naked nuclei, but did not induce internucleosomal DNA fragmentation. Finally, we demonstrated that TCR engagement of a murine T cell line (EL4) with anti-CD3/CD28 resulted in the release of Cyp B from the microsome fraction to the cytosol/nuclear fraction. Our data strongly suggest that both active caspase-activated DNase and Cyp B may participate in the induction of chromosomal DNA degradation during cell death execution of TCR-stimulated thymocytes. PMID- 11035063 TI - Arsenite induces apoptosis of murine T lymphocytes through membrane raft-linked signaling for activation of c-Jun amino-terminal kinase. AB - Because of its dual roles in acute toxicity and in therapeutic application in cancer treatment, arsenic has recently attracted a renewed attention. In this study, we report NaAsO(2)-induced signal cascades from the cell surface to the nucleus of murine thymic T lymphocytes that involve membrane rafts as an initial signal transducer. NaAsO(2) induced apoptosis through fragmentation of DNA, activation of caspase, and reciprocal regulation of Bcl-2/Bax with the concomitant reduction of membrane potential. We demonstrated that NaAsO(2) induced caspase activation is dependent on curcumin-sensitive c-Jun amino terminal kinase and barely dependent on SB203580-sensitive p38 kinase or PD98059 sensitive extracellular signal-regulated kinase. Additionally, staurosporine, which severely inhibited the activation of mitogen-activated protein (MAP) family kinases and c-Jun, partially blocked the NaAsO(2)-mediated signal for poly(ADP ribose) polymerase (PARP) degradation. Potentially as the initial cell surface event for intracellular signaling, NaAsO(2) induced aggregation of GPI-anchored protein Thy-1 and superoxide production. This Thy-1 aggregation and subsequent activation of MAP family kinase and c-Jun and the degradation of PARP induced by NaAsO(2) were all inhibited by DTT, suggesting the requirement of interaction between arsenic and protein sulfhydryl groups for those effects. beta cyclodextrin, which sequestrates cholesterol from the membrane rafts, inhibited NaAsO(2)-induced activation of protein tyrosine kinases and MAP family kinases, degradation of PARP, and production of superoxide. In addition, beta cyclodextrin dispersed NaAsO(2)-induced Thy-1 clustering. These results suggest that a membrane raft integrity-dependent cell surface event is a prerequisite for NaAsO(2)-induced protein tyrosine kinase/c-Jun amino-terminal kinase activation, superoxide production, and downstream caspase activation. PMID- 11035064 TI - Autoregulation of human monocyte-derived dendritic cell maturation and IL-12 production by cyclooxygenase-2-mediated prostanoid production. AB - PG added to cell culture profoundly affect the in vitro maturation and function of monocyte-derived dendritic cells (MDC). Because unstimulated monocytes express cyclooxygenase (COX)-1, and COX-2 when activated, we examined whether MDC express these enzymes and produce prostanoids that autoregulate maturation and IL-12 production. Immature MDC (I-MDC) and mature MDC express COX-1, but, unlike monocytes, both MDC populations constitutively express COX-2. However, COX-2 regulation in both MDC populations differs from monocytes, as IL-4 does not suppress enzyme expression. COX-2 is functional in MDC as a specific inhibitor, NS-398, significantly reduces PGE(2) production. I-MDC undergoing maturation with soluble CD40 ligand (sCD40L) increase PGE(2) synthesis, but prostanoid synthesis is switched to COX-1. However, with IFN-gamma present, sCD40L-stimulated PG metabolism is redirected to COX-2, and PGE(2) synthesis increases severalfold. Endogenous PG production by MDC does not regulate CD40, CD80, CD86, or HLA DR expression; however, it does promote MDC maturation, as NS-398 significantly reduces CD83 expression in I-MDC matured with sCD40L/IFN-gamma. PG produced through COX-2 also autoregulate IL-12, but the effects are dependent on the MDC maturation state. Blocking COX-2 reduces I-MDC secretion of IL-12p40, whereas it increases IL-12p40 and p70 production by maturing MDC. COX-2-mediated PG production impacts MDC function as maturing these cells in the presence of NS-398 yields MDC that stimulate significantly more IFN-gamma in an allogeneic mixed lymphocyte response than MDC matured without this inhibitor. These studies demonstrate that MDC express both COX isoforms constitutively and produce prostanoids, which autoregulate their maturation and function. PMID- 11035065 TI - Selective bystander proliferation of memory CD4+ and CD8+ T cells upon NK T or T cell activation. AB - Ag-experienced or memory T cells have increased reactivity to recall Ag, and can be distinguished from naive T cells by altered expression of surface markers such as CD44. Memory T cells have a high turnover rate, and CD8(+) memory T cells proliferate upon viral infection, in the presence of IFN-alphabeta and/or IL-15. In this study, we extend these findings by showing that activated NKT cells and superantigen-activated T cells induce extensive bystander proliferation of both CD8(+) and CD4(+) memory T cells. Moreover, proliferation of memory T cells can be induced by an IFN-alphabeta-independent, but IFN-gamma- or IL-12-dependent pathway. In these conditions of bystander activation, proliferating memory (CD44(high)) T cells do not derive from activation of naive (CD44(low)) T cells, but rather from bona fide memory CD44(high) T cells. Together, these data demonstrate that distinct pathways can induce bystander proliferation of memory T cells. PMID- 11035066 TI - IL-2R beta agonist P1-30 acts in synergy with IL-2, IL-4, IL-9, and IL-15: biological and molecular effects. AB - From the sequence of human IL-2 we have recently characterized a peptide (p1-30), which is the first IL-2 mimetic described. P1-30 covers the entire alpha helix A of IL-2 and spontaneously folds into a alpha helical homotetramer mimicking the quaternary structure of a hemopoietin. This neocytokine interacts with a previously undescribed dimeric form of the human IL-2 receptor beta-chain likely to form the p1-30 receptor (p1-30R). P1-30 acts as a specific IL-2Rbeta agonist, selectively inducing activation of CD8 and NK lymphocytes. From human PBMC we have also shown that p1-30 induces the activation of lymphokine-activated killer cells and the production of IFN-gamma. Here we demonstrate the ability of p1-30 to act in synergy with IL-2, -4, -9, and -15. These synergistic effects were analyzed at the functional level by using TS1beta, a murine T cell line endogenously expressing the common cytokine gamma gene and transfected with the human IL-2Rbeta gene. At the receptor level, we show that expression of human IL 2Rbeta is absolutely required to obtain synergistic effects, whereas IL-2Ralpha specifically impedes the synergistic effects obtained with IL-2. The results suggest that overexpression of IL-2Ralpha inhibits p1-30R formation in the presence of IL-2. Finally, concerning the molecular effects, although p1-30 alone induces the antiapoptotic molecule bcl-2, we show that it does not influence mRNA expression of c-myc, c-jun, and c-fos oncogenes. In contrast, p1-30 enhances IL-2 driven expression of these oncogenes. Our data suggest that p1-30R (IL-2Rbeta)(2) and intermediate affinity IL-2R (IL-2Rbetagamma), when simultaneously expressed at the cell surface, may induce complementary signal transduction pathways and act in synergy. PMID- 11035067 TI - Enhancement of T cell receptor signaling by a mild oxidative shift in the intracellular thiol pool. AB - Exposure of T cells to the macrophage products hydrogen peroxide (HP) or L lactate (LAC) was previously shown to enhance IL-2 production and to modulate glutathione (GSH) status. We now found that 50 microM HP and 30 mM LAC enhanced strongly the transcription from the IL-2 promoter in Jurkat T cells after stimulation with anti-CD28 together with or without anti-CD3 but not with anti CD3 Abs alone. Therefore, we used anti-CD3 plus anti-CD28-stimulated cells to investigate the effect of the GSH reductase inhibitor 1, 3-bis(2-chloroethyl)-1 nitrosourea (BCNU) on the signal cascade. BCNU enhanced the transcription to a similar extent as HP or LAC. Lowering the intracellular GSH/GSH disulfide ratio by BCNU, HP, or NO resulted in all cases in the fulminant enhancement of Jun-N terminal kinase and p38 mitogen-activated protein kinase but not extracellular signal-regulated kinase 1/2. Jun-N-terminal kinase and NF-kappaB activation was enhanced through pathways involving Rac, Vav1, PKCTheta, p56(lck), p59(fyn), and IkappaB kinases. In a cell-free system, the autophosphorylation of rFyn was stimulated by GSH disulfide but not by HP. These findings suggest that the oxidation of the cellular thiol pool may play a role as an amplifying mechanism for TCR/CD3 signals in immune responses. PMID- 11035068 TI - IL-15 and IL-15 receptor selectively regulate differentiation of common mucosal immune system-independent B-1 cells for IgA responses. AB - We show in this report a new regulatory role for IL-15 and IL-15R in the development of B-1 cells and their differentiation into IgA-producing cells. Mucosal IgA levels were found to be inhibited by anti-IL-15 mAb treatment in vivo, but enhanced by administration of rIL-15, while serum IgA levels remained unaffected. Mucosal B-1 cells preferentially proliferated in response to IL-15 in vitro. When mucosal B-1 and B-2 cells were separated into surface (s)IgM(+)sIgA( ) and sIgM(-)sIgA(+) fractions, IL-15R-specific mRNA was found to be predominant in both sIgM(+)sIgA(-) and sIgM(-)sIgA(+) B-1 cells at a much higher level than B 2 cells. Further, incubation of these different subsets of B-1 and B-2 cells with IL-15 resulted in greater enhancement of the corresponding receptor expression by B-1 subset when compared with B-2 fraction. Interestingly, de novo isolated sIgM(+)sIgA(-) B-1, but not sIgM(+)sIgA(-) B-2, cells were already class-switched cells because the germline Calpha transcript was detected and was then further enhanced by IL-15. IL-15 also supported differentiation of both sIgM(+)sIgA(-) and sIgM(-)sIgA(+) B-1 cells into IgA-producing cells. Taken together, these findings suggest that IL-15 is a critically important cytokine for the differentiation of both sIgM(+),IgA(-) and sIgM(-)sIgA(+) B-1 cells expressing IL 15R into IgA-producing cells in mucosal tissues. PMID- 11035069 TI - Maturation-dependent expression and function of the CD49d integrin on monocyte derived human dendritic cells. AB - Dendritic cells (DC) are highly specialized APC that are critical for the initiation of T cell-dependent immune responses. DC exert a sentinel function while immature and, after activation by inflammatory stimuli or infectious agents, mature and migrate into lymphoid organs to prime T cells. We have analyzed integrin expression on monocyte-derived DC (MDDC) and found that expression of CD49d integrins (CD49d/CD29 and CD49d/beta7) was induced/up regulated during TNF-alpha- or LPS-initiated MDDC maturation, reflecting the induction/up-regulation of CD49d and beta7 mRNA. CD49d mRNA steady-state level increased more than 10 times during maturation, with the highest levels observed 24 h after TNF-alpha treatment. CD49d integrin expression conferred mature MDDC with an elevated capacity to adhere to the CS-1 fragment of fibronectin, and also mediated transendothelial migration of mature MDDC. Up-regulation of CD49d integrin expression closely paralleled that of the mature DC marker CD83. CD49d integrin expression was dependent on cell maturation, as its induction was abrogated by N:-acetylcysteine, which inhibits NF-kappaB activation and the functional and phenotypic maturation of MDDC. Moreover, CD49d integrin up regulation and MDDC maturation were prevented by SB203580, a specific inhibitor of p38 mitogen-activated protein kinase, but were almost unaffected by the mitogen-activated protein/extracellular signal-related kinase kinase 1/2 inhibitor PD98059. Our results support the existence of a link between functional and phenotypic maturation of MDDC and CD49d integrin expression, thus establishing CD49d as a maturation marker for MDDC. The differential expression of CD49d on immature and mature MDDC might contribute to their distinct motility capabilities and mediate mature DC migration into lymphoid organs. PMID- 11035070 TI - The expression of p18INK4 and p27kip1 cyclin-dependent kinase inhibitors is regulated differently during human B cell differentiation. AB - Cell cycle progression is under the control of cyclin-dependent kinases (cdks), the activity of which is dependent on the expression of specific cdk inhibitors. In this paper we report that the two cdk inhibitors, p27(Kip1) and p18(INK4c), are differently expressed and control different steps of human B lymphocyte activation. Resting B cells contain large amounts of p27(Kip1) and no p18(INK4c). In vitro stimulation by Staphylococcus aureus Cowan 1 strain or CD40 ligand associated with IL-10 and IL-2 induces a rapid decrease in p27(Kip1) expression combined with cell cycle entry and progression. In contrast, in vitro Ig production correlates with specific expression of p18(INK4c) and early G(1) arrest. This G(1) arrest is associated with inhibition of cyclin D3/cdk6-mediated retinoblastoma protein phosphorylation by p18(INK4c). A similar contrasting pattern of p18(INK4c) and p27(Kip1) expression is observed both in B cells activated in vivo and in various leukemic cells. Expression of p18(INK4c) was also detected in various Ig-secreting cell lines in which both maximum Ig secretion and specific p18(INK4c) expression were observed during the G(1) phase. Our study shows that p27(Kip1) and p18(INK4c) have different roles in B cell activation; p27(Kip1) is involved in the control of cell cycle entry, and p18(INK4c) is involved in the subsequent early G(1) arrest necessary for terminal B lymphocyte differentiation. PMID- 11035071 TI - Unexpected autoantibody production in membrane Ig-mu-deficient/lpr mice. AB - In the B lymphocyte lineage, Fas-mediated cell death is important in controlling activated mature cells, but little is known about possible functions at earlier developmental stages. In this study we found that in mice lacking the IgM transmembrane tail exons (muMT mice), in which B cell development is blocked at the pro-B stage, the absence of Fas or Fas ligand allows significant B cell development and maturation, resulting in high serum Ig levels. These B cells demonstrate Ig heavy chain isotype switching and autoimmune reactivity, suggesting that lack of functional Fas allows maturation of defective and/or self reactive B cells in muMT/lpr mice. Possible mechanisms that may allow maturation of these B cells are discussed. PMID- 11035072 TI - Induction and expression of beta-calcitonin gene-related peptide in rat T lymphocytes and its significance. AB - Our previous data have shown that rat lymphocytes can synthesize calcitonin gene related peptide (CGRP), a neuropeptide. In this study the type, characteristics, and functional role of lymphocyte-derived CGRP were investigated. The results showed that treatment with Con A (4 microg/ml) and recombinant human IL-2 (rhIL 2; 750 U/ml) for 3-5 days induced CGRP synthesis and secretion by lymphocytes from both thymus and mesenteric lymph nodes in a time-dependent manner. Stimulation of these cells with Con A (1-8 microg/ml) or rhIL-2 (94-1500 U/ml) for 5 days induced a significant increase in CGRP secretion in a concentration dependent manner. The maximal secretion of CGRP with Con A by thymocytes was elevated from 104+/-11 to 381 +/- 44 pg/10(8) cells, and that by mesenteric lymph node lymphocytes was elevated from 83+/-10 to 349+/-25 pg/10(8) cells, respectively. The maximal CGRP secretion with rhIL-2 by thymocytes was elevated from 116+/-3 to 607+/-23 pg/10(8), and that by mesenteric lymph node lymphocytes was elevated from 117+/-9 to 704+/- 37 pg/10(8) cells, respectively. The nucleotide sequencing study showed that lymphoid cells expressed beta-CGRP cDNA only. The levels of beta-CGRP mRNA in mitogen-stimulated lymphocytes of both sources were also increased. However, LPS had no such effect on either source of cells. hCGRP(8-37) (2.0 microM), a CGRP(1) receptor antagonist, enhanced Con A induced proliferation and IL-2 release of thymocytes by 41.3 and 35.8% over those induced by Con A alone, respectively. The data suggest that T lymphocyte mitogens can induce the production of endogenous beta-CGRP from T lymphocytes, which may partially inhibit the proliferation and IL-2 release of rat T lymphocyte under immune challenges. PMID- 11035073 TI - Fas/Fas ligand interactions promote activation-induced cell death of NK T lymphocytes. AB - NKT cells are a versatile population whose immunoregulatory functions are modulated by their microenvironment. We demonstrate herein that in addition to their IFN-gamma production, NKT lymphocytes stimulated with IL-12 plus IL-18 in vitro underwent activation in terms of CD69 expression, blast transformation, and proliferation. Yet they were unable to survive in culture because, once activated, they were rapidly eliminated by apoptosis, even in the presence of their survival factor IL-7. This process was preceded by up-regulation of Fas (CD95) and Fas ligand expression in response to IL-12 plus IL-18 and was blocked by zVAD, a large spectrum caspase inhibitor, as well as by anti-Fas ligand mAb, suggesting the involvement of the Fas pathway. In accordance with this idea, NKT cells from Fas-deficient C57BL/6-lpr/lpr mice did not die in these conditions, although they shared the same features of cell activation as their wild-type counterpart. Activation-induced cell death occurred also after TCR engagement in vivo, since NKT cells became apoptotic after injection of their cognate ligand, alpha-galactosylceramide, in wild-type, but not in Fas-deficient, mice. Taken together, our data provide the first evidence for a new Fas-dependent mechanism allowing the elimination of TCR-dependent or -independent activated NKT cells, which are potentially dangerous to the organism. PMID- 11035074 TI - CXCR4 receptor expression on human retinal pigment epithelial cells from the blood-retina barrier leads to chemokine secretion and migration in response to stromal cell-derived factor 1 alpha. AB - Retinal pigment epithelial (RPE) cells form part of the blood-retina barrier and have recently been shown to produce various chemokines in response to proinflammatory cytokines. As the scope of chemokine action has been shown to extend beyond the regulation of leukocyte migration, we have investigated the expression of chemokine receptors on RPE cells to determine whether they could be a target for chemokine signaling. RT-PCR analysis indicated that the predominant receptor expressed on RPE cells was CXCR4. The level of CXCR4 mRNA expression, but not cell surface expression, increased on stimulation with IL-1beta or TNF alpha. CXCR4 protein could be detected on the surface of 16% of the RPE cells using flow cytometry. Calcium mobilization in response to the CXCR4 ligand stromal cell-derived factor 1alpha (SDF-1alpha) indicated that the CXCR4 receptors were functional. Incubation with SDF-1alpha resulted in secretion of monocyte chemoattractant protein-1, IL-8, and growth-related oncogene alpha. RPE cells also migrated in response to SDF-1alpha. As SDF-1alpha expression by RPE cells was detected constitutively, we postulate that SDF-1-CXCR4 interactions may modulate the affects of chronic inflammation and subretinal neovascularization at the RPE site of the blood-retina barrier. PMID- 11035075 TI - Distinct human T cell repertoires mediate immediate and delayed-type hypersensitivity to the Trichophyton antigen, Tri r 2. AB - The 29-kDa subtilase homologue, Tri r 2, derived from the dermatophyte fungus Trichophyton rubrum, exhibits unique immunologic characteristics in its ability to elicit immediate (IH) and delayed-type (DTH) hypersensitivity skin tests in different individuals. Thus, Tri r 2 provides a model for comparing the T cell repertoire in subjects with distinct immune responses to a single Ag. Recombinant Tri r 2 produced as a GST fusion protein in Escherichia coli stimulated strong in vitro lymphoproliferative responses in 10 IH and 10 DTH responders. Patterns of T cell epitope recognition were compared between skin test groups using 28 overlapping peptides (each in 12 replicate wells) derived from Tri r 2 to stimulate T lymphocyte proliferation in vitro. Peptide 5 (P5; aa 41-60) induced the strongest response in DTH subjects and showed the largest difference between DTH and IH responders in proliferation (mean standardized index, 2.22 and 0.82, respectively; p = 0.0047) and number of positive wells (81 vs 12). Responses to P5 were associated with diverse HLA haplotypes. These results showed that P5 contains an immunodominant epitope specifically associated with DTH and that this peptide is recognized in a permissive manner. Cross-validated linear discriminant analysis using T cell proliferative responses to two regions of Tri r 2 (aa 51-90 and 231-270) gave a 95% predictive accuracy for classification of subjects into IH or DTH groups. We conclude that different immune responses to Trichophyton are mediated by distinct T cell repertoires between individuals with IH and DTH reactions to Tri r 2. PMID- 11035076 TI - Macrophage-derived dendritic cells have strong Th1-polarizing potential mediated by beta-chemokines rather than IL-12. AB - Monocyte-derived dendritic cells (MDDCs) activate naive T lymphocytes to induce adaptive immunity, effecting Th1 polarization through IL-12. However, little is known about other potential DC Th1 polarizing mechanisms, or how T cell polarization may be affected by DCs differentiating in, or exposed to, a proinflammatory environment. Macrophages (MPhis) are DC precursors abundant in inflamed tissues, lymph nodes, and tumors. Thus we studied the T cell-activating and -polarizing properties of MPhi-derived DCs (PhiDCs). Monocytes were cultured in MPhi-CSF (M-CSF) to produce MPhis, which were then differentiated into DCs following culture with GM-CSF plus IL-4. PhiDCs activated a significant allogeneic MLR and were significantly better than MDDCs in activating T cells with superantigen. Most strikingly, PhiDCs elicited up to 9-fold more IFN-gamma from naive or Ag-specific T cells compared with MDDCs (with equivalent IL-4 secretion), despite producing up to 9-fold less IL-12. Neutralization of MDDC, but not PhiDC IL-12 significantly inhibited T cell IFN-gamma induction. PhiDCs produced up to 12-fold more beta-chemokines (macrophage-inflammatory protein 1alpha, -1beta, and RANTES) than MDDCs. Ab blockade of CCR5, but not CXC chemokine receptor 4, inhibited T cell IFN-gamma induction by PhiDCs significantly greater than by MDDCs. Thus DCs differentiating from MPhis induce T cell IFN-gamma through beta-chemokines with little or no requirement for IL-12. Myeloid DCs arising from distinct precursor cells may have differing properties, including different mechanisms of Th1 polarization. These data are the first reports of IFN-gamma induction through chemokines by DCs. PMID- 11035077 TI - Reciprocal expression of the TNF family receptor herpes virus entry mediator and its ligand LIGHT on activated T cells: LIGHT down-regulates its own receptor. AB - The TNF receptor (TNFR) family plays a central role in the development of the immune response. Here we describe the reciprocal regulation of the recently identified TNFR superfamily member herpes virus entry mediator (HVEM) (TR2) and its ligand LIGHT (TL4) on T cells following activation and the mechanism of this process. T cell activation resulted in down-regulation of HVEM and up-regulation of LIGHT, which were both more pronounced in CD8(+) than CD4(+) T lymphocytes. The analysis of HVEM and LIGHT mRNA showed an increase in the steady state level of both mRNAs following stimulation. LIGHT, which was present in cytoplasm of resting T cells, was induced both in cytoplasm and at the cell surface. For HVEM, activation resulted in cellular redistribution, with its disappearance from cell surface. HVEM down-regulation did not rely on de novo protein synthesis, in contrast to the partial dependence of LIGHT induction. Matrix metalloproteinase inhibitors did not modify HVEM expression, but did enhance LIGHT accumulation at the cell surface. However, HVEM down-regulation was partially blocked by a neutralizing mAb to LIGHT or an HVEM-Fc fusion protein during activation. As a model, we propose that following stimulation, membrane or secreted LIGHT binds to HVEM and induces receptor down-regulation. Degradation or release of LIGHT by matrix metalloproteinases then contributes to the return to baseline levels for both LIGHT and HVEM. These results reveal a self-regulating ligand/receptor system that contributes to T cell activation through the interaction of T cells with each other and probably with other cells of the immune system. PMID- 11035078 TI - Decreased neutrophil adhesion to human cytomegalovirus-infected retinal pigment epithelial cells is mediated by virus-induced up-regulation of Fas ligand independent of neutrophil apoptosis. AB - Human CMV (HCMV) retinitis frequently leads to blindness in iatrogenically immunosuppressed patients and in the end stage of AIDS. Despite the general proinflammatory potential of HCMV, virus infection is associated with a rather mild cellular inflammatory response in the retina. To investigate this phenomenon, the influence of HCMV (strains AD169 or Hi91) infection on C-X-C chemokine secretion, ICAM-1 expression, and neutrophil recruitment in cultured human retinal pigment epithelial (RPE) cells was studied. Supernatants from infected cultures contained enhanced levels of IL-8 and melanoma growth stimulating activity/Gro alpha and induced neutrophil chemotaxis compared with supernatants from uninfected RPE cells. Despite HCMV-induced ICAM-1 expression on RPE cells, binding of activated neutrophils to HCMV-infected RPE cells and subsequent transepithelial penetration were significantly reduced. Reduced neutrophil adhesion to infected RPE cells correlated with HCMV-induced up regulation of constitutive Fas ligand (FasL) expression. Functional blocking of FasL on RPE cells with the neutralizing mAbs NOK-1 and NOK-2 or of the Fas receptor on neutrophils with mAbB-D29 prevented the HCMV-induced impairment of neutrophil/RPE interactions. Fas-FasL-dependent impairment of neutrophil binding had occurred by 10 min after neutrophil/RPE coculture without apoptotic signs. Neutrophil apoptosis was first detected after 4 h. Treatment of neutrophils with a specific inhibitor of caspase-8 suppressed apoptosis, whereas it did not prevent impaired neutrophil binding to infected RPE. The current results suggest a novel role for FasL in the RPE regulation of neutrophil binding. This may be an important feature of virus escape mechanisms and for sustaining the immune privileged character of the retina during HCMV ocular infection. PMID- 11035079 TI - Identification of five different Patr class I molecules that bind HLA supertype peptides and definition of their peptide binding motifs. AB - We have sequenced the Pan troglodytes class I (Patr) molecules from three common chimpanzees and expressed them as single molecules in a class I-deficient cell line. These lines were utilized to obtain purified class I molecules to define the peptide binding motifs associated with five different Patr molecules. Based on these experiments, as well as analysis of the predicted structure of the B and F polymorphic MHC pockets, we classified five Patr molecules (Patr-A*0101, Patr B*0901, Patr-B*0701, Patr-A*0602, and Patr-B*1301) within previously defined supertype specificities associated with HLA class I molecules (HLA-A3, -B7, -A1, and -A24 supertypes). The overlap in the binding repertoire between specific HLA and Patr class I molecules was in the range of 33 to 92%, depending on the particular Patr molecule as assessed by the binding of HIV-, hepatitis B virus-, and hepatitis C virus-derived epitopes. Finally, live cell binding assays of nine chimpanzee-derived B cell lines demonstrated that HLA supertype peptides bound to Patr class I molecules with frequencies in the 20-50% range. PMID- 11035080 TI - Molecular cloning, characterization, and expression of TNF cDNA and gene from Japanese flounder Paralychthys olivaceus. AB - We cloned a cDNA and the gene for Japanese flounder TNF. The TNF cDNA consisted of 1217 bp, which encoded 225 amino acid residues. The identities between Japanese flounder TNF and members of the mammalian TNF family were approximately 20-30%. The positions of cysteine residues that are important for disulfide bonds were conserved with respect to those in mammalian TNF-alpha. The Japanese flounder TNF gene has a length of approximately 2 kbp and consists of four exons and three introns. The positions of the exon-intron junction positions of Japanese flounder TNF gene are similar to those of human TNF-alpha. However, the length of the first intron of Japanese flounder is much shorter than that of the human TNF-alpha gene. There are simple CA or AT dinucleotide repeats in the 5' upstream and 3'-downstream regions of the Japanese flounder TNF gene. Southern blot hybridization indicted that Japanese flounder TNF exists as a single copy. Expression of Japanese flounder TNF mRNA is greatly induced after stimulation of PBLs with LPS, Con A, or PMA. These results indicated that Japanese flounder TNF is more like mammalian TNF-alpha than mammalian lymphotoxin-alpha, with respect to its gene structure, length of amino acid sequence, number and position of cysteine residues, and regulation of gene expression. PMID- 11035081 TI - Notch signaling enhances survival and alters differentiation of 32D myeloblasts. AB - The Notch transmembrane receptors play important roles in precursor survival and cell fate specification during hematopoiesis. To investigate the function of Notch and the signaling events activated by Notch in myeloid development, we expressed truncated forms of Notch1 or Notch2 proteins that either can or cannot activate the core binding factor 1 (CBF1) in 32D (clone 3) myeloblasts. 32D cells proliferate as blasts in the presence of the cytokines, GM-CSF or IL-3, but they initiate differentiation and undergo granulopoiesis in the presence of granulocyte CSF (G-CSF). 32D cells expressing constitutively active forms of Notch1 or Notch2 proteins that signal through the CBF1 pathway maintained significantly higher numbers of viable cells and exhibited less cell death during G-CSF induction compared with controls. They also displayed enhanced entry into granulopoiesis, and inhibited postmitotic terminal differentiation. In contrast, Notch1 constructs that either lacked sequences necessary for CBF1 binding or that failed to localize to the nucleus had little effect. Elevated numbers of viable cells during G-CSF treatment were also observed in 32D cells overexpressing the basic helix-loop-helix protein (bHLH), HES1, consistent with activation of the CBF1 pathway. Taken together, our data suggest that Notch signaling enhances 32D cell survival, promotes entry into granulopoiesis, and inhibits postmitotic differentiation through a CBF1-dependent pathway. PMID- 11035082 TI - Cell-specific expression of the murine CD21 gene depends on accessibility of promoter and intronic elements. AB - The murine complement receptor type 2 gene (Cr2/CD21) is transcriptionally active in murine B and follicular dendritic cells, but not in murine T cells. We have previously shown that altering chromatin structure via histone deacetylase inhibitors results in CD21 expression in murine T cells, and that the minimal CD21 promoter provided appropriate cell-specific expression of luciferase reporter constructs only in the presence of the first third of intron 1, fragment A. We extend this work by showing that replacing the CD21 gene promoter with the SV40 promoter resulted in the loss of this cell-specific control. Further delineation of intronic regulatory elements by fragmentation also resulted in the loss of cell-specific gene expression, suggesting that multiple CD21 promoter and intronic elements interact for appropriate CD21 gene expression. To assess this model, we performed EMSAs to define protein binding sites within promoter and intronic regions and DNase I hypersensitivity assays to determine chromatin accessibility. Multiple DNA binding factors were shown to be present in B and T cell extracts; a minority demonstrated B cell specificity. However, the DNase I sensitivity of T cell CD21 regulatory elements was not comparable to that of B cells until the histone acetylation status of the gene was altered. Taken together, these data suggest that chromatin remodeling facilitates cell-specific CD21 gene expression by modulating access of transcription factors to regulatory elements in the promoter and intron. PMID- 11035083 TI - Identification and characterization of a beta proteasome subunit cluster in the Japanese pufferfish (Fugu rubripes). AB - The low molecular mass polypeptide (LMP2, LMP7, and MECL-1) genes code for beta type subunits of the proteasome, a multimeric complex that degrades proteins into peptides as part of the MHC class I-mediated Ag-presenting pathway. These gene products are up-regulated in response to infection by IFN-gamma and replace the corresponding constitutively expressed subunits (X, Y, and Z) during the immune response. In humans, the LMP2 and LMP7 genes both reside within the class II region of the MHC (6p21.3), while MECL-1 is located at 16q22.1. In the present study, we have identified all three IFN-gamma-regulated beta-type proteasome subunits in Fugu, which are present as a cluster within the Fugu MHC class I region. We show that in this species, LMP7, LMP2, and MECL-1 are linked. Also within this cluster is an LMP2-like subunit (which seems specific to all teleosts tested to date) and a closely linked LMP7 pseudogene, indicating that within Fugu and potentially other teleosts, there has been an additional regional duplication involving these genes. PMID- 11035084 TI - Mutational analysis reveals multiple distinct sites within Fc gamma receptor IIB that function in inhibitory signaling. AB - The low-affinity receptor for IgG, FcgammaRIIB, functions broadly in the immune system, blocking mast cell degranulation, dampening the humoral immune response, and reducing the risk of autoimmunity. Previous studies concluded that inhibitory signal transduction by FcgammaRIIB is mediated solely by its immunoreceptor tyrosine-based inhibition motif (ITIM) that, when phosphorylated, recruits the SH2-containing inositol 5'- phosphatase SHIP and the SH2-containing tyrosine phosphatases SHP-1 and SHP-2. The mutational analysis reported here reveals that the receptor's C-terminal 16 residues are also required for detectable FcgammaRIIB association with SHIP in vivo and for FcgammaRIIB-mediated phosphatidylinositol 3-kinase hydrolysis by SHIP. Although the ITIM appears to contain all the structural information required for receptor-mediated tyrosine phosphorylation of SHIP, phosphorylation is enhanced when the C-terminal sequence is present. Additionally, FcgammaRIIB-mediated dephosphorylation of CD19 is independent of the cytoplasmic tail distal from residue 237, including the ITIM. Finally, the findings indicate that tyrosines 290, 309, and 326 are all sites of significant FcgammaRIIB1 phosphorylation following coaggregation with B cell Ag receptor. Thus, we conclude that multiple sites in FcgammaRIIB contribute uniquely to transduction of FcgammaRIIB-mediated inhibitory signals. PMID- 11035085 TI - Metalloprotease-mediated shedding of enzymatically active mouse ecto-ADP ribosyltransferase ART2.2 upon T cell activation. AB - T cells proteolytically shed the ectodomains of several cell surface proteins and, thereby, can alter their responsiveness and can release soluble intercellular regulators. ART2.2 is a GPI-anchored ecto-ADP-ribosyltransferase (ART) related to ADP-ribosylating bacterial toxins. ART2.2 is expressed exclusively by mature T cells. Here we show that ART2.2 is shed from the cell surface in enzymatically active form upon activation of T cells. Shedding of ART2.2 resembles that of L-selectin (CD62L) in dose response, kinetics of release, and sensitivity to the metalloprotease inhibitor Immunex Compound 3, suggesting that ART2.2, like CD62L, is cleaved by TNF-alpha-converting enzyme or by another metalloprotease. ART2.2 shed from activated T cells migrates slightly faster in SDS-PAGE analyses than does ART2.2 released upon cleavage of the GPI anchor. This indicates that shedding of ART2.2 is mediated by proteolytic cleavage close to its membrane anchor. Shed ART2.2 is enzymatically active and ADP-ribosylates several substrates in vitro. Thus, shedding of ART2.2 releases a potential intercellular regulator. Finally, using a new FACS assay for monitoring ADP-ribosylation of cell surface proteins, we demonstrate that shedding of ART2.2 correlates with a reduced sensitivity of T cell surface proteins to ADP ribosylation. Our findings suggest that by shedding ART2.2 the activated T cell not only releases a potential intercellular regulator but also may alter its responsiveness to immune regulation by ART2.2-mediated ADP-ribosylation of cell surface proteins. PMID- 11035086 TI - Regulated production and molecular diversity of human liver and activation regulated chemokine/macrophage inflammatory protein-3 alpha from normal and transformed cells. AB - Liver and activation-regulated chemokine (LARC), also designated macrophage inflammatory protein-3alpha (MIP-3alpha), Exodus, or CCL20, is a C-C chemokine that attracts immature dendritic cells and memory T lymphocytes, both expressing CCR6. Depending on the cell type, this chemokine was found to be inducible by cytokines (IL-1beta) and by bacterial, viral, or plant products (including LPS, dsRNA, and PMA) as measured by a specific ELISA. Although coinduced with monocyte chemotactic protein-1 (MCP-1) and IL-8 by dsRNA, measles virus, and IL-1beta in diploid fibroblasts, leukocytes produced LARC/MIP-3alpha only in response to LPS. However, in myelomonocytic THP-1 cells LARC/MIP-3alpha was better induced by phorbol ester, whereas in HEp-2 epidermal carcinoma cells IL-1beta was the superior inducer. The production levels of LARC/MIP-3alpha (1-10 ng/ml) were, on the average, 10- to 100-fold lower than those of IL-8 and MCP-1, but were comparable to those of other less abundantly secreted chemokines. Natural LARC/MIP-3alpha protein isolated from stimulated leukocytes or tumor cell lines showed molecular diversity, in that NH(2)- and COOH-terminally truncated forms were purified and identified by amino acid sequence analysis and mass spectrometry. In contrast to other chemokines, including MCP-1 and IL-8, the natural processing did not affect the calcium-mobilizing capacity of LARC/MIP 3alpha through its receptor CCR6. Furthermore, truncated natural LARC/MIP-3alpha isoforms were equally chemotactic for lymphocytes as intact rLARC/MIP-3alpha. It is concluded that in addition to its role in homeostatic trafficking of leukocytes, LARC/MIP-3alpha can function as an inflammatory chemokine during host defense. PMID- 11035087 TI - Identification of a complex that binds to the CD154 3' untranslated region: implications for a role in message stability during T cell activation. AB - CD154 expression is regulated throughout a time course of CD3-dependent T cell activation by differential mRNA decay. To understand the molecular basis of the "stability" phase of this pathway, experiments were conducted to identify sequences and specific complexes important in this regulation. Gel retardation assays using extracts from both Jurkat T cells and CD3-activated CD4(+) T cells revealed a major complex (complex I) that bound a 65-bp highly CU-rich region of the CD154 3' untranslated region. The specificity of the CU-rich element for complex-I formation was confirmed by disruption of this complex by oligo(dCT) competition. Formation of complex I strongly correlated with CD154 mRNA stability across a time course of T cell activation. UV cross-linking identified a major oligo(dCT)-sensitive species at approximately 90 kDa that showed induced and increased expression in extracts from 24- and 48-hr anti-CD3-activated T cells, respectively. This protein was absent in equivalent extracts from resting or 2-h activated T cells. Using an in vitro decay assay, we found that a CD154-specific transcript was more rapidly degraded in 2-h-activated extract and stabilized in the 24- and 48-h extracts compared to extracts from resting T cells. Disruption of complex I resulted in the rapid decay of a CD154-specific transcript demonstrating a functional role for complex I in mRNA stabilization in vitro. These studies support a model of posttranscriptional regulation of CD154 expression being controlled in part by the interaction of a poly(CU)-binding complex with a specific sequence in the 3' untranslated region. PMID- 11035088 TI - Heavy chain revision in MRL mice: a potential mechanism for the development of autoreactive B cell precursors. AB - Abs reactive to DNA and DNA/histone complexes are distinguished by the presence of positively charged amino acids, such as arginine, in the heavy chain complementarity-determining region 3. The presence of these amino acids partly results from atypical V(H)-D-J(H) rearrangements such as D-D fusions and D inversions. Previous results in our laboratory demonstrated that newborn autoimmune MRL/MpJ-+/+ mice undergo these unusual recombinations more frequently when compared with normal C3H/HeJ controls. In addition, the heavy chain junctions in newborn MRL mice demonstrated a preferred usage of V(H)-proximal D genes and distal J(H) genes suggestive of secondary gene rearrangements. In this study we explore the possibility that adult MRL B220(+)IgM(-) pre B cells, which have not yet undergone Ag selection, exhibit similar rearrangement patterns. Indeed, MRL pre-B cells possessed more atypical rearrangements (D-D fusions) than those of C3H/HeJ mice. However, the biased use of upstream D genes and downstream J(H) genes observed in the newborn MRL mice was not present in the pre-B cell library. These results suggest that the heavy chain rearrangement process persists later during B cell life in lupus-prone mice and lead us to propose a model of heavy chain receptor revision in the periphery of autoimmune mice. PMID- 11035089 TI - Molecular recognition of human CD1b antigen complexes: evidence for a common pattern of interaction with alpha beta TCRs. AB - Ag-specific T cell recognition is mediated through direct interaction of clonotypic TCRs with complexes formed between Ag-presenting molecules and their bound ligands. Although characterized in substantial detail for class I and class II MHC encoded molecules, the molecular interactions responsible for TCR recognition of the CD1 lipid and glycolipid Ag-presenting molecules are not yet well understood. Using a panel of epitope-specific Abs and site-specific mutants of the CD1b molecule, we showed that TCR interactions occur on the membrane distal aspects of the CD1b molecule over the alpha1 and alpha2 domain helices. The location of residues on CD1b important for this interaction suggested that TCRs bind in a diagonal orientation relative to the longitudinal axes of the alpha helices. The data point to a model in which TCR interaction extends over the opening of the putative Ag-binding groove, making multiple direct contacts with both alpha helices and bound Ag. Although reminiscent of TCR interaction with MHC class I, our data also pointed to significant differences between the TCR interactions with CD1 and MHC encoded Ag-presenting molecules, indicating that Ag receptor binding must be modified to accommodate the unique molecular structure of the CD1b molecule and the unusual Ags it presents. PMID- 11035090 TI - Changing the antigen binding specificity by single point mutations of an anti-p24 (HIV-1) antibody. AB - The murine mAb CB4-1 raised against p24 (HIV-1) recognizes a linear epitope of the HIV-1 capsid protein. Additionally, CB4-1 exhibits cross-reactive binding to epitope-homologous peptides and polyspecific reactions to epitope nonhomologous peptides. Crystal structures demonstrate that the epitope peptide (e-pep) and the nonhomologous peptides adopt different conformations within the binding region of CB4-1. Site-directed mutagenesis of the fragment variable (Fv) region was performed using a single-chain (sc)Fv construct of CB4-1 to analyze binding contributions of single amino acid side chains toward the e-pep and toward one epitope nonhomologous peptide. The mutations of Ab amino acid side chains, which are in direct contact with the Ag, show opposite influences on the binding of the two peptides. Whereas the affinity of the e-pep to the CB4-1 scFv mutant heavy chain variable region Tyr(32)Ala is decreased 250-fold, the binding of the nonhomologous peptide remains unchanged. In contrast, the mutation light chain variable region Phe(94)Ala reduces the affinity of the nonhomologous peptide 10 fold more than it does for the e-pep. Thus, substantial changes in the specificity can be observed by single amino acid exchanges. Further characterization of the scFv mutants by substitutional analysis of the peptides demonstrates that the effect of a mutation is not restricted to contact residues. This method also reveals an inverse compensatory amino acid exchange for the nonhomologous peptide which increases the affinity to the scFv mutant light chain variable region Phe(94)Ala up to the level of the e-pep affinity to the wild-type scFv. PMID- 11035091 TI - Rapid recruitment of neutrophils containing prestored IL-12 during microbial infection. AB - Neutrophils are well known to rapidly migrate to foci of infection, where they exert microbicidal functions. We sought to determine whether neutrophils responding to in vivo infection with the protozoan pathogen Toxoplasma gondii were capable of IL-12 production as suggested by recent in vitro studies. Intraperitoneal infection induced a neutrophil influx by 4 h, accompanied by ex vivo IL-12 p40 and p70 release. Approximately 85% of the neutrophils displayed intracellular stores of IL-12, as determined by flow cytometry and confocal fluorescence microscopy. Neutrophils from IFN-gamma knockout mice also expressed IL-12, ruling out an IFN-gamma-priming requirement. Neither infected nor uninfected peritoneal macrophages displayed intracellular IL-12, but these cells were strongly IL-10(+). Infection per se was unnecessary for IL-12 production because peritoneal and peripheral blood neutrophils from uninfected animals contained IL-12(+) populations. Expression of the granulocyte maturation marker Gr-1 (Ly-6G) was correlated with IL-12 production. Mice depleted of their granulocytes by mAb administration at the time of infection had decreased serum levels of IL-12 p40. These results suggest a model in which neutrophils with prestored IL-12 are rapidly mobilized to an infection site where they are triggered by the parasite to release cytokine. Our findings place neutrophils prominently in the cascade of early events leading to IL-12-dependent immunity to T. gondii. PMID- 11035092 TI - Adenovirus E1A oncogene expression in tumor cells enhances killing by TNF-related apoptosis-inducing ligand (TRAIL). AB - Expression of the adenovirus serotype 5 (Ad5) E1A oncogene sensitizes cells to apoptosis by TNF-alpha and Fas-ligand. Because TNF-related apoptosis-inducing ligand (TRAIL) kills cells in a similar manner as TNF-alpha and Fas ligand, we asked whether E1A expression might sensitize cells to lysis by TRAIL. To test this hypothesis, we examined TRAIL-induced killing of human melanoma (A2058) or fibrosarcoma (H4) cells that expressed E1A following either infection with Ad5 or stable transfection with Ad5-E1A. E1A-transfected A2058 (A2058-E1A) or H4 (H4 E1A) cells were highly sensitive to TRAIL-induced killing, but Ad5-infected cells expressing equally high levels of E1A protein remained resistant to TRAIL. Infection of A2058-E1A cells with Ad5 reduced their sensitivity to TRAIL dependent killing. Therefore, viral gene products expressed following infection with Ad5 inhibited the sensitivity to TRAIL-induced killing conferred by transfection with E1A. E1B and E3 gene products have been shown to inhibit TNF alpha- and Fas-dependent killing. The effect of these gene products on TRAIL dependent killing was examined by using Ad5-mutants that did not express either the E3 (H5dl327) or E1B-19K (H5dl250) coding regions. A2058 cells infected with H5dl327 were susceptible to TRAIL-dependent killing. Furthermore, TRAIL-dependent killing of A2058-E1A cells was not inhibited by infection with H5dl327. Infection with H5dl250 sensitized A2058 cells to TRAIL-induced killing, but considerably less than H5dl327-infection. In summary, expression of Ad5-E1A gene products sensitizes cells to TRAIL-dependent killing, whereas E3 gene products, and to a lesser extent E1B-19K, inhibit this effect. PMID- 11035093 TI - Novel mechanism of antibody-independent complement neutralization of herpes simplex virus type 1. AB - The envelope surface glycoprotein C (gC) of HSV-1 interferes with the complement cascade by binding C3 and activation products C3b, iC3b, and C3c, and by blocking the interaction of C5 and properdin with C3b. Wild-type HSV-1 is resistant to Ab independent complement neutralization; however, HSV-1 mutant virus lacking gC is highly susceptible to complement resulting in > or =100-fold reduction in virus titer. We evaluated the mechanisms by which complement inhibits HSV-1 gC null virus to better understand how gC protects against complement-mediated neutralization. C8-depleted serum prepared from an HSV-1 and -2 Ab-negative donor neutralized gC null virus comparable to complement-intact serum, indicating that C8 and terminal lytic activity are not required. In contrast, C5-depleted serum from the same donor failed to neutralize gC null virus, supporting a requirement for C5. EDTA-treated serum did not neutralize gC null virus, indicating that complement activation is required. Factor D-depleted and C6-depleted sera neutralized virus, suggesting that the alternative complement pathway and complement components beyond C5 are not required. Complement did not aggregate virus or block attachment to cells. However, complement inhibited infection before early viral gene expression, indicating that complement affects one or more of the following steps in virus replication: virus entry, uncoating, DNA transport to the nucleus, or immediate early gene expression. Therefore, in the absence of gC, HSV-1 is readily inhibited by complement by a C5-dependent mechanism that does not require viral lysis, aggregation, or blocking virus attachment. PMID- 11035094 TI - Increased resistance against acute polymicrobial sepsis in mice challenged with immunostimulatory CpG oligodeoxynucleotides is related to an enhanced innate effector cell response. AB - Recent reports support the concept that the major defect in polymicrobial sepsis is an impaired immunologic response to infection. Oligodeoxynucleotides containing CpG sequence motifs (CpG-ODN) were previously shown to induce immune protection in models of chronic infection with intracellular bacteria, parasites, and viruses due to their ability to augment IFN-gamma-dependent Th1 responses. Here, we demonstrate that challenging mice with CpG-ODN substantially increases the resistance against acute polymicrobial sepsis. Systemic levels of IL-12, IL 18, and IL-10 were not altered in CpG-ODN-treated mice as compared with controls. In contrast, administration of CpG-ODN resulted in a strongly enhanced accumulation of neutrophils at the primary site of infection. Neutrophils of CpG ODN-treated mice exhibited an up-regulation of phagocytic receptors, an increased phagocytic activity, and an elevated production of reactive oxygen metabolites. These results suggest that the protective effects of CpG-ODNs in acute polymicrobial sepsis are related to an enhanced effector cell response of innate immunity. CpG-ODN may therefore represent potent agents for the treatment of sepsis-associated immunoparalysis. PMID- 11035095 TI - Role of IL-5 in innate and adaptive immunity to larval Strongyloides stercoralis in mice. AB - Protective immunity to Strongyloides stercoralis infective larvae in mice has been shown to be dependent on IL-5 based on mAb depletion studies. The goal of this study was to determine the functional role of IL-5 during the innate and adaptive immune response to larval S. stercoralis in mice. In these studies, three strains of mice were used: wild-type C57BL/6J (WT), IL-5 knockout (KO), and IL-5 transgenic (TG). Innate responses to the larvae indicated that there was enhanced survival in the KO animals and decreased survival in the TG animals compared with WT. Furthermore, killing of larvae in TG mice was associated with eosinophil infiltration and degranulation. In studying the adaptive immune response, it was observed that immunization of KO mice did not lead to the development of protective immunity. Experiments were then performed to determine whether KO mice reconstituted with Abs or cells could then develop protective immunity. KO mice displayed protective immunity via a granulocyte-dependent mechanism following injection of purified IgM from immune wild-type animals. Immunity in KO mice could also be reconstituted by the injection of eosinophils at the time of immunization. These eosinophils did not participate in actively killing the challenge infection, but rather were responsible for the induction of a protective Ab response. We conclude that IL-5 is required in the protective immune response for the production of eosinophils, and that eosinophils were involved in larval killing during innate immunity and in the induction of protective Abs in the adaptive immune response. PMID- 11035096 TI - Memory-type CD8+ T cells protect IL-2 receptor alpha-deficient mice from systemic infection with herpes simplex virus type 2. AB - IL-2Ralpha-deficient (IL-2Ralpha(-/-)) mice exhibit an impaired activation induced cell death for T cells and develop abnormal T cell activation with age. In our study, we found that IL-2Ralpha(-/-) mice at the age of 5 wk contained an increased number of CD44(+)CD69(-)CD8(+) T cells in lymph nodes, which expressed a high intensity of IL-2Rbeta and vigorously proliferated in response to a high dose of IL-15 or IL-2. The T cells produced a large amount of IFN-gamma in response to IL-15 plus IL-12 in a TCR-independent bystander manner. When IL 2Ralpha(-/-) mice were inoculated i.p. with HSV type 2 (HSV-2) 186 strain, they showed resistance to the infection accompanied by an increased level of serum IL 15. The depletion of CD8(+) T cells by in vivo administration of anti-CD8 mAb rendered IL-2Ralpha(-/-) mice susceptible to HSV-2-induced lethality. These results suggest that memory-type CD8(+) T cells play a novel role in the protection against HSV-2 infection in IL-2Ralpha(-/-) mice. PMID- 11035097 TI - Lipopolysaccharide-binding protein is vectorially secreted and transported by cultured intestinal epithelial cells and is present in the intestinal mucus of mice. AB - Lipopolysaccharide-binding protein (LBP) is an important modulator of the host's response to endotoxin. In a previous study, we found evidence for the synthesis of LBP by intestinal epithelial cells. In this study, we explored the polarity of LBP secretion by these cells. Polarized monolayers of Caco-2 cells were used as intestinal mucosa model. Cells were stimulated apically or basally with cytokines, and LBP secretion was analyzed. Furthermore, the presence of LBP in intestinal mucus of healthy and endotoxemic mice was studied using a mucus sampling technique. The constitutive unipolar LBP secretion from the apical cell surface was markedly enhanced when cells were exposed to cytokines at their apical surface. However, bioactive LBP was secreted from both cell surfaces after basolateral stimulation of cells. Cytokines also influenced the secretion of the acute phase proteins serum amyloid A, apoA-I, and apoB from both surfaces of Caco 2 cells. Furthermore, transport of exogenous LBP from the basolateral to the apical cell surface was demonstrated. In line with these in vitro data, the presence of LBP in intestinal mucus was strongly enhanced in mice after a challenge with endotoxin. The results indicate that LBP is present at the mucosal surface of the intestine, a phenomenon for which secretion and transport of LBP by intestinal epithelial cells may be responsible. PMID- 11035098 TI - A role for parasite-induced PGE2 in IL-10-mediated host immunoregulation by skin stage schistosomula of Schistosoma mansoni. AB - Significant quantities of PGE(2) were produced by cercariae of Schistosoma mansoni following incubation with linoleic acid, a free fatty acid found on the surface of the skin. Cyclooxygenase (COX) 2 inhibitors failed to block this PGE(2) production, suggesting that a different biochemical pathway may be involved in the production of PGE(2) by the parasite. In addition, the parasites were also able to induce PGE(2) and IL-10 from human and mouse keratinocytes. Analysis of mouse skin during skin migratory phases of infection confirmed these in vitro observations. COX2 inhibitors blocked the parasite-induced PGE(2) and IL 10 from keratinocytes. Further analysis of the parasite secretions showed that the PGE(2)/IL-10-inducing effect was associated with a fraction <30 kDa molecular size. Addition of this fraction or parasite-stimulated keratinocyte culture supernatant to Con A-stimulated spleen cells resulted in the suppression of cell proliferation. This effect could be blocked by anti-IL-10 treatment. In sharp contrast, attenuation of the parasites with gamma-irradiation significantly abrogated their ability to induce PGE(2) or IL-10 from skin cells. Significance of IL-10 in host immunoregulation by skin stage schistosomula of S. mansoni was further confirmed by using IL-10-deficient mice. In these mice the normal subdued cutaneous reaction to the parasite was absent. Instead, a prominent cellular reaction occurred around the parasite, and there was considerable delay in parasitic migration through the skin. Thus these results suggest a key role for parasite-induced PGE(2) in IL-10-dependent down-regulation of host immune responses in the skin. PMID- 11035099 TI - Partially TAP-independent protection against Listeria monocytogenes by H2-M3 restricted CD8+ T cells. AB - Effective protection against Listeria monocytogenes requires Ag-specific CD8(+) T cells. A substantial proportion of CD8(+) T cells activated during L. monocytogenes infection of C57BL/6 mice are restricted by the MHC class Ib molecule H2-M3. In this study, an H2-M3-restricted CD8(+) T cell clone specific for a known H2-M3 epitope (fMIGWII) was generated from L. monocytogenes-infected mice. The clone was cytotoxic, produced IFN-gamma, and could mediate strong protection against L. monocytogenes when transferred to infected mice. Macrophages pulsed with heat-killed LISTERIAE: presented Ag to the clone in a TAP independent manner. Both TAP-independent and -dependent processing occurred in vivo, as TAP-deficient mice infected with L. monocytogenes were partially protected by adoptive transfer of the clone. This is the first example of CD8(+) T cell-mediated, TAP-independent protection against a pathogen in vivo, confirming the importance of alternative MHC class I processing pathways in the antibacterial immunity. PMID- 11035100 TI - Induction of vigorous helper and cytotoxic T cell as well as B cell responses by dendritic cells expressing a modified antigen targeting receptor-mediated internalization pathway. AB - Efficient Ag presentation is essential to induce effective cellular and humoral immune responses. Thus, one central goal of current immunotherapy and vaccine development is to enhance Ag presentation to induce potent and broad immune responses. Here, a novel Ag presentation strategy is developed by transducing dendritic cells (DCs) to produce an Ag for presentation as an exogenous Ag to efficiently induce both humoral and cellular immunity. The principle of this strategy is illustrated by genetically modifying DCs to secrete a model hepatitis B virus Ag fused with a cell-binding domain and to process the fusion Ag as an exogenous Ag after receptor-mediated internalization for MHC class I and II presentation. Vigorous Ag-specific CD4(+) helper and CD8(+) cytotoxic T cell, as well as B cell, responses were induced by the transduced DCs in mouse models. Thus, this novel strategy uses a receptor-mediated internalization process to efficiently induce all arms of the adaptive immunity and may provide a powerful means to develop potent vaccines and immunotherapies. PMID- 11035101 TI - The Rel family member P50 mediates cytokine-induced C-reactive protein expression by a novel mechanism. AB - Transcription of C-reactive protein (CRP) in Hep 3B cells is induced by IL-6, acting through C/EBP isoforms and STAT3. IL-1beta, which alone has no effect, greatly enhances IL-6-induced transcription by unknown mechanisms. Because IL 1beta activates the NF-kappaB system, we explored the effects of overexpressed Rel family members on CRP expression. Unexpectedly, transactivation assays in transiently transfected Hep 3B cells showed p50 overexpression to markedly induce CRP transcription, acting in a region 3' to -86. In the presence of overexpressed p50, IL-1beta induced a 3-fold increase in CRP expression, and responses to IL-6 and to IL-6 plus IL-1beta were 4-fold greater than seen in cells without p50 overexpression. In contrast, overexpressed p65 abolished CRP induction by p50 and by cytokines. EMSA studies demonstrated that recombinant p50 bound to a nonconsensus kappaB site overlapping the proximal C/EBP binding site on the CRP promoter. Mutation of a polypyrimidine tract in the p50-binding site inhibited the transactivating effect of cytokines. P50- but not p65-containing dimers were found in nuclei of Hep 3B cells 18 h after stimulation with IL-1beta, when C/EBPbeta is greatly activated, in the presence or absence of IL-6. These findings suggest that IL-1beta induces nuclear translocation of p50-containing dimers and that p50 interacts with C/EBPbeta activated by both IL-6 and IL-1beta to induce CRP expression. PMID- 11035102 TI - The synthetic peptide Trp-Lys-Tyr-Met-Val-D-Met is a potent chemotactic agonist for mouse formyl peptide receptor. AB - Formyl peptides are potent neutrophil chemoattractants. In humans and rabbits, the formyl peptide receptor (FPR) binds N-formyl-Met-Leu-Phe (fMLF) with high affinity (K(d) approximately 1 nM). The mouse FPR (mFPR) is a low-affinity receptor for fMLF (K(d) approximately 100 nM); therefore, other agonists for this receptor may exist. Using mFPR-transfected rat basophilic leukemia cells, we found that a recently identified synthetic peptide Trp-Lys-Tyr-Met-Val-D-Met (WKYMVm) is a potent agonist for mFPR. WKYMVm induced calcium mobilization with an EC(50) of 1.2-1.5 nM. Optimal chemotaxis was achieved with 1 nM of WKYMVm, but it required 100 nM of fMLF. WKYMVm stimulated rapid and potent phosphorylation of the mitogen-activated protein kinases extracellular signal-related kinases 1 and 2 when used at 50 nM. Pertussis toxin only partially blocked calcium mobilization and production of inositol 1,4,5-trisphosphate in the stimulated mFPR cells, suggesting the possibility that this receptor couples to Galpha proteins other than Gi and Go. Competitive binding and desensitization data suggest that both peptides interact with the same receptor but may use nonoverlapping binding sites because WKYMVm was unable to effectively displace [(3)H]fMLF bound to mFPR. These results provide evidence for the presence of an alternative potent agonist for mFPR, and suggest a potential usage of WKYMVm for probing the ligand-receptor interactions with the murine formyl peptide receptor homologs. PMID- 11035103 TI - Role of intracellular chloride in the reversible activation of neutrophil beta 2 integrins: a lesson from TNF stimulation. AB - The process of beta(2) integrin activation, which enhances the interaction of these heterodimers with ligands, plays a crucial role in the adherence-dependent neutrophilic polymorphonuclear leukocytes' (PMN) responses to TNF. Our previous observation, showing that a marked decrease of the high basal Cl(-) content (Cl( )(i)) is an essential step in the TNF-induced activation of PMN, stimulated this study, which investigates the role of alterations of Cl(-)(i) in the activation of beta(2) integrins triggered by TNF. Here we show that TNF enhances the expression of activation-specific neoepitopes of beta(2) integrins, namely, epitope 24, a unique epitope present on all three leukocyte integrin alpha subunits, and epitope CBRM1/5, localized to the I domain on the alpha-chain of Mac-1 (CD11bCD18). Moreover, we demonstrate that the conformational changes underlying the expression of the neoepitopes are dependent on a drop in Cl(-)(i) because 1) inhibition of Cl(-)(i) decrease is invariably accompanied by inhibition of beta(2) integrin activation, 2) Cl(-)(i) decrease induced by means other than agonist stimulation, i.e., by placing PMN in Cl(-)-free buffers, activates beta(2) integrins, and 3) restoration of the original Cl(-)(i) levels is accompanied by deactivation of beta(2) integrins. We also show that Cl(-)(i) decrease is required for TNF-induced cytoplasmic alkalinization, but such a rise in pH(i) does not seem to be relevant for beta(2) integrin activation. The results of our study emphasize the role of Cl(-) as a new PMN "second messenger." PMID- 11035104 TI - ATP acts as an agonist to promote stimulus-induced secretion of IL-1 beta and IL 18 in human blood. AB - Cultured monocytes and macrophages stimulated with LPS produce large quantities of proIL-1beta, but release little mature cytokine to the medium. The efficiency at which the procytokine is converted to its active 17-kDa species and released extracellularly is enhanced by treating cytokine-producing cells with a secretion stimulus such as ATP or nigericin. To determine whether this need for a secretion stimulus extends to blood, individual donors were bled twice daily for 4 consecutive days, and the collected blood samples were subjected to a two-step IL 1 production assay. LPS-activated blood samples generated cell-free IL-1beta, but levels of the extracellular cytokine were greatly increased by subsequent treatment with ATP or nigericin. Specificity and concentration requirements of the nucleotide triphosphate effect suggests a P2X(7) receptor involvement. Quantities of IL-1beta generated by an individual donor's blood in response to the LPS-only and LPS/ATP stimuli were relatively consistent over the 4-day period. Between donors, consistent differences in cytokine production capacity were observed. Blood samples treated with ATP also demonstrated enhanced IL-18 production, but TNF-alpha levels decreased. Among leukocytes, monocytes appeared to be the most affected cellular targets of the ATP stimulus. These studies indicate that an exogenous stimulus is required by blood for the efficient production of IL-1beta and IL-18, and suggest that circulating blood monocytes constitutively express a P2X(7)-like receptor. PMID- 11035105 TI - CD47 ligation selectively inhibits the development of human naive T cells into Th1 effectors. AB - The CD47 Ag, also named integrin-associated protein, was recently reported to regulate the production of IL-12 by human monocytes and dendritic cells. The present study shows that CD47 ligation by CD47 mAb in primary cultures of cord blood mononuclear cells inhibits IL-12-driven Th1 cell development, as revealed by the cytokine secretion profile at restimulation and IFN-gamma production at the single-cell level. F(ab')(2) fragments of CD47 mAb or the synthetic peptide 4N1K, corresponding to the CD47 binding site of thrombospondin, display the same activity. CD47 engagement does not change the phenotype of IL-12-primed cells from Th1 to Th2 or affect IL-4-induced Th2 cell development. Moreover, CD47 mAb inhibits IL-12- but not IL-4-induced IL-2 production as well as IFN-gamma in primary cultures, which was correlated with a decrease of the IL-12Rbeta2 chain expression. Inclusion of exogenous IL-2 at priming corrects IL-12R expression as well as the inhibition of Th1 cell development. The data thus underline the role of IL-2 in Th1 cell development and further suggest that targeting IL-2 and IL-12 simultaneously may have some therapeutic advantage in Th1 autoimmune diseases. PMID- 11035106 TI - Protein kinase C zeta plays a central role in activation of the p42/44 mitogen activated protein kinase by endotoxin in alveolar macrophages. AB - Human alveolar macrophages respond to endotoxin (LPS) by activation of a number of mitogen-activated protein kinase pathways, including the p42/44 (extracellular signal-related kinase (ERK)) kinase pathway. In this study, we evaluated the role of the atypical protein kinase C (PKC) isoform, PKC zeta, in LPS-induced activation of the ERK kinase pathway. Kinase activity assays showed that LPS activates PKC zeta, mitogen-activated protein/ERK kinase (MEK, the upstream activator of ERK), and ERK. LPS did not activate Raf-1, the classic activator of MEK. Pseudosubstrate-specific peptides with attached myristic acid are cell permeable and can be used to block the activity of specific PKC isoforms in vivo. We found that a peptide specific for PKC zeta partially blocked activation of both MEK and ERK by LPS. We also found that this peptide blocked in vivo phosphorylation of MEK after LPS treatment. In addition, we found that LPS caused PKC zeta to bind to MEK in vivo. These observations suggest that MEK is an LPS directed target of PKC zeta. PKC zeta has been shown in other systems to be phosphorylated by phosphatidylinositol (PI) 3-kinase-dependent kinase. We found that LPS activates PI 3-kinase and causes the formation of a PKC zeta/PI 3-kinase dependent kinase complex. These data implicate the PI 3-kinase pathway as an integral part of the LPS-induced PKC zeta activation. Taken as a whole, these studies suggest that LPS activates ERK kinase, in part, through activation of an atypical PKC isoform, PKC zeta. PMID- 11035107 TI - In situ amplification of 5-lipoxygenase and 5-lipoxygenase-activating protein in allergic airway inflammation and inhibition by leukotriene blockade. AB - Leukotrienes are important mediators of the eosinophilic influx and mucus hypersecretion in the lungs in a murine model of asthma. We used in situ PCR in this model of human asthma to detect lung mRNA for 5-lipoxygenase (5-LO) and 5-LO activating protein (FLAP), key proteins necessary for leukotriene synthesis. Lung tissue was obtained on day 28 from mice treated with i.p. (days 0 and 14) and intranasal (days 14, 25, 26, and 27) OVA or saline. After fixation, the tissue sections underwent protease- and RNase-free DNase digestion, before in situ RT PCR using target-specific cDNA amplification. 5-LO and FLAP-specific mRNA was visualized by a digoxigenin detection system, and positive cells were analyzed by morphometry. 5-LO and FLAP-specific mRNA and protein were associated primarily with eosinophils and alveolar macrophages in the airways and pulmonary blood vessels in OVA-sensitized/challenged mice. 5-LO and FLAP protein expression increased on a per-cell basis in alveolar macrophages of OVA-treated mice compared with saline controls. Pulmonary blood vessel endothelial cells were also positive for 5-LO, FLAP mRNA, and protein. 5-LO inhibition significantly decreased 5-LO and FLAP-specific mRNA and protein expression in the lung inflammatory cells and endothelial cells. These studies demonstrate a marked increase in key 5-LO pathway proteins in the allergic lung inflammatory response and an important immunomodulatory effect of leukotriene blockade to decrease 5-LO and FLAP gene expression. PMID- 11035108 TI - IL-18 has IL-12-independent effects in delayed-type hypersensitivity: studies in cell-mediated crescentic glomerulonephritis. AB - IL-18 (formerly known as IFN-gamma-inducing factor) enhances Th1 responses via effects that are thought to be dependent on and synergistic with IL-12. The potential for IL-18 to exert IL-12-independent effects in delayed-type hypersensitivity (DTH) responses was studied in a model of Th1-directed, DTH mediated crescentic glomerulonephritis induced by planting an Ag in glomeruli of sensitized mice as well as in cutaneous DTH. Sensitized genetically normal (IL 12(+/+)) mice developed proteinuria and crescentic glomerulonephritis with a glomerular influx of DTH effectors (CD4(+) T cells, macrophages, and fibrin deposition) in response to the planted glomerular Ag. IL-12p40-deficient (IL-12( /-)) mice showed significant reductions in crescent formation, proteinuria, and glomerular DTH effectors. Administration of IL-18 to IL-12(-/-) mice restored the development of histological (including effectors of DTH) and functional glomerular injury in IL-12(-/-) mice to levels equivalent to those in IL-12(+/+) mice. IL-18 administration to IL-12(-/-) mice increased glomerular ICAM-1 protein expression, but did not restore Ag-stimulated splenocyte IFN-gamma, GM-CSF, IL-2, or TNF-alpha production. Sensitized IL-12(+/+) mice also developed cutaneous DTH following intradermal challenge with the nephritogenic Ag. Cutaneous DTH was inhibited in IL-12(-/-) mice, but was restored by administration of IL-18. IL 12(+/+) mice given IL-18 developed augmented injury, with enhanced glomerular and cutaneous DTH, demonstrating the synergistic effects of IL-18 and IL-12 in DTH responses. These studies demonstrate that even in the absence of IL-12, IL-18 can induce in vivo DTH responses and up-regulate ICAM-1 without inducing IFN-gamma, GM-CSF, or TNF-alpha production. PMID- 11035109 TI - ICAM-1-induced expression of proinflammatory cytokines in astrocytes: involvement of extracellular signal-regulated kinase and p38 mitogen-activated protein kinase pathways. AB - ICAM-1 is a transmembrane glycoprotein of the Ig superfamily involved in cell adhesion. ICAM-1 is aberrantly expressed by astrocytes in CNS pathologies such as multiple sclerosis, experimental allergic encephalomyelitis, and Alzheimer's disease, suggesting a possible role for ICAM-1 in these disorders. ICAM-1 has been shown to be important for leukocyte diapedesis through brain microvessels and subsequent binding to astrocytes. However, other functional roles for ICAM-1 expression on astrocytes have not been well elucidated. Therefore, we investigated the intracellular signals generated upon ICAM-1 engagement on astrocytes. ICAM-1 ligation by a mAb to rat ICAM-1 induced mRNA expression of proinflammatory cytokines such as IL-1alpha, IL-1beta, IL-6, and TNF-alpha. Examination of cytokine protein production revealed that ICAM-1 ligation results in IL-6 secretion by astrocytes, whereas IL-1beta and IL-1alpha protein is expressed intracellularly in astrocytes. The involvement of mitogen-activated protein kinases (MAPKs) in ICAM-1-mediated cytokine expression in astrocytes was tested, as the MAPK extracellular signal-regulated kinase (ERK) was previously shown to be activated upon ICAM-1 engagement. Our results indicate that ERK1/ERK2, as well as p38 MAPK, are activated upon ligation of ICAM-1. Studies using pharmacological inhibitors demonstrate that both p38 MAPK and ERK1/2 are involved in ICAM-1-induced IL-6 expression, whereas only ERK1/2 is important for IL-1alpha and IL-1beta expression. Our data support the role of ICAM-1 on astrocytes as an inflammatory mediator in the CNS and also uncover a novel signal transduction pathway through p38 MAPK upon ICAM-1 ligation. PMID- 11035110 TI - In vivo roles of integrins during leukocyte development and traffic: insights from the analysis of mice chimeric for alpha 5, alpha v, and alpha 4 integrins. AB - Mice chimeric for integrins alpha(5), alpha(V), or alpha(4) were used to dissect the in vivo roles of these adhesion receptors during leukocyte development and traffic. No major defects were observed in the development of lymphocytes, monocytes, or granulocytes or in the traffic of lymphocytes to different lymphoid organs in the absence of alpha(5) or alpha(V) integrins. However, in agreement with previous reports, the absence of alpha(4) integrins produced major defects in development of lymphoid and myeloid lineages and a specific defect in homing of lymphocytes to Peyer's patches. In contrast, the alpha(4) integrin subunit is not essential for localization of T lymphocytes into intraepithelial and lamina propria compartments in the gut, whereas one of the partners of alpha(4), the beta(7) chain, has been shown to be essential. However, alpha(4)-deficient T lymphocytes cannot migrate properly during the inflammatory response induced by thioglycolate injection into the peritoneum. Finally, in vitro proliferation and activation of lymphocytes deficient for alpha(5), alpha(V), or alpha(4) integrins upon stimulation with different stimuli were similar to those seen in controls. These results show that integrins play distinct roles during in vivo leukocyte development and traffic. PMID- 11035111 TI - Human monoclonal antibodies isolated from type I diabetes patients define multiple epitopes in the protein tyrosine phosphatase-like IA-2 antigen. AB - Protein tyrosine phosphatase-like IA-2 autoantigen is one of the major targets of humoral autoimmunity in patients with insulin-dependant diabetes mellitus (IDDM). In an effort to define the epitopes recognized by autoantibodies against IA-2, we generated five human mAbs (hAbs) from peripheral B lymphocytes isolated from patients most of whom had been recently diagnosed for IDDM. Determination and fine mapping of the critical regions for autoantibody binding was performed by RIA using mutant and chimeric constructs of IA-2- and IA-2beta-regions. Four of the five IgG autoantibodies recognized distinct epitopes within the protein tyrosine phosphatase (PTP)-like domain of IA-2. The minimal region required for binding by three of the PTP-like domain-specific hAbs could be located to aa 777 979. Two of these hAbs cross-reacted with the related IA-2beta PTP-like domain (IA-2beta aa 741-1033). A further PTP-like domain specific hAb required the entire PTP-like domain (aa 687-979) for binding, but critical amino acids clustered in the N-terminal region 687-777. An additional epitope could be localized within the juxtamembrane domain (aa 603-779). In competition experiments, the epitope recognized by one of the hAbs was shown to be targeted by 10 of 14 anti-IA-2-positive sera. Nucleotide sequence analysis of this hAb revealed that it used a V(H) germline gene (DP-71) preferably expressed in autoantibodies associated with IDDM. The presence of somatic mutations in both heavy and light chain genes and the high affinity or this Ab suggest that the immune response to IA-2 is Ag driven. PMID- 11035113 TI - The soluble endothelial protein C receptor binds to activated neutrophils: involvement of proteinase-3 and CD11b/CD18. AB - The protein C pathway is a primary regulator of blood coagulation and a critical component of the host response to inflammatory stimuli. The most recent member of this pathway is the endothelial protein C receptor (EPCR), a type I transmembrane protein with homology to CD1d/MHC class I proteins. EPCR accelerates formation of activated protein C, a potent anticoagulant and antiinflammatory agent. The current study demonstrates that soluble EPCR binds to PMA-activated neutrophils. Using affinity chromatography, binding studies with purified components, and/or blockade with specific Abs, it was found that soluble EPCR binds to proteinase-3 (PR3), a neutrophil granule proteinase. Furthermore, soluble EPCR binding to neutrophils was partially dependent on Mac-1 (CD11b/CD18), a beta(2) integrin involved in neutrophil signaling, and cell-cell adhesion events. PR3 is involved in multiple diverse processes, including hemopoietic proliferation, antibacterial activity, and autoimmune-mediated vasculitis. The observation that soluble EPCR binds to activated neutrophils via PR3 and a beta(2) integrin suggests that there may be a link between the protein C anticoagulant pathway and neutrophil functions. PMID- 11035112 TI - Impaired CD4 T cell activation due to reliance upon B cell-mediated costimulation in nonobese diabetic (NOD) mice. AB - Diabetes in nonobese diabetic (NOD) mice results from the activation of I-A(g7) restricted, islet-reactive T cells. This study delineates several characteristics of NOD CD4 T cell activation, which, independent of I-A(g7), are likely to promote a dysregulated state of peripheral T cell tolerance. NOD CD4 T cell activation was found to be resistant to antigenic stimulation via the TCR complex, using the progression of cell division as a measure. The extent of NOD CD4 T cell division was highly sensitive to changes in Ag ligand density. Moreover, even upon maximal TCR complex-mediated stimulation, NOD CD4 T cell division prematurely terminated. Maximally stimulated NOD CD4 T cells failed to achieve the threshold number of division cycles required for optimal susceptibility to activation-induced death, a critical mechanism for the regulation of peripheral T cell tolerance. Importantly, these aberrant activation characteristics were not T cell-intrinsic but resulted from reliance on B cell costimulatory function in NOD mice. Costimulation delivered by nonautoimmune strain APCs normalized NOD CD4 T cell division and the extent of activation induced death. Thus, by disrupting the progression of CD4 T cell division, polarization of APC costimulatory function to the B cell compartment could allow the persistence and activation of diabetogenic cells in NOD mice. PMID- 11035114 TI - Extracellular ATP and TNF-alpha synergize in the activation and maturation of human dendritic cells. AB - Extracellular ATP mediates numerous biological activities by interacting with plasma membrane P2 purinergic receptors. Recently, P2 receptors have been described on dendritic cells (DC), but their functional role remains unclear. Proposed functions include improved Ag presentation, cytokine production, chemotaxis, and induction of apoptosis. We investigated the effects of ATP and of other P2 receptor agonists on endocytosis, phenotype, IL-12 secretion, and T cell stimulatory capacity of human monocyte-derived DC. We found that in the presence of extracellular ATP, DC transiently increase their endocytotic activity. Subsequently, DC up-regulate CD86, CD54, and MHC-II; secrete IL-12; and exhibit an improved stimulatory capacity for allogeneic T cells. These effects were more pronounced when chemically modified ATP derivatives with agonistic activity on P2 receptors, which are resistent to degradation by ectonucleotidases, were applied. Furthermore, ATP and TNF-alpha synergized in the activation of DC. Stimulated with a combination of ATP and TNF-alpha, DC expressed the maturation marker CD83, secreted large amounts of IL-12, and were potent stimulators of T cells. In the presence of the P2 receptor antagonist suramin, the effects of ATP were completely abolished. Our results suggest that extracellular ATP may play an important immunomodulatory role by activating DC and by skewing the immune reaction toward a Th1 response through the induction of IL-12 secretion. PMID- 11035115 TI - HIV gag mRNA transfection of dendritic cells (DC) delivers encoded antigen to MHC class I and II molecules, causes DC maturation, and induces a potent human in vitro primary immune response. AB - Dendritic cells (DC) are the major APCs involved in naive T cell activation making them prime targets of vaccine research. We observed that mRNA was efficiently transfected, resulting in superior translation in DC compared with other professional APCs. A single stimulation of T cells by HIV gag-encoded mRNA transfected DC in vitro resulted in primary CD4(+) and CD8(+) T cell immune responses at frequencies of Ag-specific cells (5-12.5%) similar to primary immune responses observed in vivo in murine models. Additionally, mRNA transfection also delivered a maturation signal to DC. Our results demonstrated that mRNA-mediated delivery of encoded Ag to DC induced potent primary T cell responses in vitro. mRNA transfection of DC, which mediated efficient delivery of antigenic peptides to MHC class I and II molecules, as well as delivering a maturation signal to DC, has the potential to be a potent and effective anti-HIV T cell-activating vaccine. PMID- 11035116 TI - Role of IL-18 in CD4+ T lymphocyte activation in sarcoidosis. AB - Sarcoidosis is a granulomatous disease of unknown etiology associated with the expansion of IL-2-producing activated CD4(+) T lymphocytes. A number of factors including the recently described IL-18 have been implicated in IL-2 expression in vitro. We investigated the role of IL-18 in IL-2 expression in sarcoidosis. Eighteen individuals with sarcoidosis and 15 normal controls were studied. IL-18R expression and epithelial lining fluid (ELF) concentrations of IL-18 were significantly elevated in the sarcoid group (p = 0.0143 and 0.0024, respectively). Both AP1 and NF-kappaB, transcription factors that regulate IL-2 gene expression, were activated in vivo in sarcoid pulmonary CD4(+) T lymphocytes. Transcription factor activity was not detected in pulmonary CD4(+) T lymphocytes from normal controls or from peripheral blood CD4(+) T lymphocytes from individuals with sarcoidosis, further evidence of compartmentalization of the lymphoproliferative process in this condition. We examined the effects of IL 18 on AP1 and NF-kappaB in Jurkat T cells in vitro. These effects were both time and dose dependent. Examination of transcription factor activation and IL-2 gene expression in Jurkat T cells revealed that sarcoid but not normal ELF activated AP1 and NF-kappaB, induced IL-2 gene transcription, and up-regulated IL-2 protein production. Addition of IL-18 to normal ELF also induced IL-2 mRNA accumulation, whereas correspondent depletion of IL-18 from sarcoid ELF using neutralizing Abs abrogated all of the effects. These data strongly implicate IL-18 in the pathogenesis of sarcoidosis via activation of AP1 and NF-kappaB, leading to enhanced IL-2 gene expression and IL-2 protein production and concomitant T cell activation. PMID- 11035117 TI - Impaired antibody affinity maturation process characterizes a subset of patients with common variable immunodeficiency. AB - Common variable immunodeficiency (CVID) is an heterogeneous syndrome characterized by decreased levels of serum Ig and recurrent bacterial infection. Here, we were interested to study whether a qualitative defect of the affinity Ab maturation process could be combined to the low level of serum Ig in a cohort of 38 CVID patients. For this, we designed a novel and rapid screening test for the detection of hypomutated V gene expressed by memory B cells. This test delineated a subset of 9/38 (23%) CVID patients with an abnormal pattern of Ig V gene mutation. The mean frequency of V gene mutation of this subset was significantly lower (1.74%) compared with other CVID patients (5.46%) and normal donors (6.5%) (p<0.0001). The mean age of this subgroup was significantly higher than other hypogammaglobulinemic patients with normal levels of V gene mutation (p<0.02), whereas no difference in the duration of symptoms was noted between the two groups. This suggests that hypomutation characterizes patients who began CVID late in life. Recently, it was shown that non-Ig sequences, such as the intronic BCL-6 gene, could be the target of the somatic hypermutation process in normal memory B cells. Our finding of a normal mutation frequency of the BCL-6 gene in two hypomutated CVID point to a defect of the Ig targeting of hypermutation machinery in these cases. PMID- 11035118 TI - In vitro and in vivo induction of a Th cell response toward peptides of the melanoma-associated glycoprotein 100 protein selected by the TEPITOPE program. AB - The melanoma-associated Ag glycoprotein 100 was analyzed by the T cell epitope prediction software TEPITOPE. Seven HLA-DR promiscuous peptides predicted with a stringent threshold were used to load dendritic cells (DC), and induction of a proliferative response was monitored. PBMC of all nine donors including two patients with malignant melanoma responded to at least one of the peptides. The proliferative response was defined as a Th response by the selective expansion of CD4(+) cells, up-regulation of CD25 and CD40L, and IL-2 and IFN-gamma expression. Peptide-loaded DC also initiated a T helper response in vivo (i.e., tumor growth in the SCID mouse was significantly retarded by the transfer of PBMC together with peptide-loaded DC). Because the use of the TEPITOPE program allows for a prediction of T cell epitopes; because the predicted peptides can be rapidly confirmed by inducing a Th response in the individual patient; and because application of peptide-loaded DC suffices for the in vivo activation of helper cells, vaccination with MHC class II-binding peptides of tumor-associated Ags becomes a feasible and likely powerful tool in the immunotherapy of cancer. PMID- 11035119 TI - Telomerase activity is increased and telomere length shortened in T cells from blood of patients with atopic dermatitis and psoriasis. AB - We studied telomerase activity and telomere length in PBMC and purified CD4(+) and CD8(+) T cells from blood obtained from a total of 32 patients with atopic dermatitis, 16 patients with psoriasis, and 30 normal controls. The telomerase activity was significantly increased in PBMC from the patients compared with PBMC from normal donors. This increase was most pronounced in the subpopulation of CD4(+) T cells, which were significantly above the activity of the CD8(+) T cells in atopic dermatitis, psoriasis patients, and control persons. The telomere length was significantly reduced in all T cell subsets from both atopic dermatitis and psoriasis patients compared with normal individuals. Furthermore, the telomere length was found to be significantly shorter in CD4(+) memory T cells compared with the CD4(+) naive T cells, and both of the cell subsets from diseases were shown to be of significantly shorter telomere length than the same cell subsets from normal controls. No significant difference was observed between CD8(+)CD28(-) and CD8(+)CD28(+) T cell populations in both diseases. However, the telomere length of CD8(+)CD28(+) T cells from both diseases was significantly shorter than CD8(+)CD28(+) T cell subsets from normal donors. In conclusion, the increased telomerase activity and shortened telomere length indicates that T lymphocytes in atopic dermatitis and psoriasis are chronically stimulated and have an increased cellular turnover in vivo. PMID- 11035120 TI - Multiepitopic HLA-A*0201-restricted immune response against hepatitis B surface antigen after DNA-based immunization. AB - CTL together with anti-envelope Abs represent major effectors for viral clearance during hepatitis B virus (HBV) infection. The induction of strong cytotoxic and Ab responses against the envelope proteins after DNA-based immunization has been proposed as a promising therapeutic approach to mediate viral clearance in chronically infected patients. Here, we studied the CTL responses against previously described hepatitis B surface Ag (HBsAg)-HLA-A*0201-restricted epitopes after DNA-based immunization in HLA-A*0201 transgenic mice. The animal model used was Human Human D(b) (HHD) mice, which are deficient for mouse MHC class I molecules (beta(2)-microglobulin(-/-) D(b-/-)) and transgenic for a chimeric HLA-A*0201/D(b) molecule covalently bound to the human beta(2) microglobulin (HHD(+/+)). Immunization of these mice with a DNA vector encoding the small and the middle HBV envelope proteins carrying HBsAg induced CTL responses against several epitopes in each animal. This study performed on a large number of animals described dominant epitopes with specific CTL induced in all animals and others with a weaker frequency of recognition. These results confirmed the relevance of the HHD transgenic mouse model in the assessment of vaccine constructs for human use. Moreover, genetic immunization of HLA-A2 transgenic mice generates IFN-gamma-secreting CD8(+) T lymphocytes specific for endogenously processed peptides and with recognition specificities similar to those described during self-limited infection in humans. This suggests that responses induced by DNA immunization could have the same immune potential as those developing during natural HBV infection in human patients. PMID- 11035121 TI - Male osteoporosis. PMID- 11035122 TI - Occupation and upper limb disorders. PMID- 11035123 TI - HLA association with autoimmune disease: a failure to protect? AB - That certain HLA specificities are associated with predisposition to autoimmune disease does not necessarily imply that self-reactive T cells restricted to particular HLA alleles are eliciting the disease. In the present essay, we argue that HLA can be a major genetic factor in the development of autoimmune diseases without T cells being primarily involved in its initiation or perpetuation. There is now ample evidence that self-reactive, regulatory T cells can protect against pernicious autoimmunity. Hereafter, we propose that extended HLA haplotypes, such as DQ3-DR4, DQ3-DR9, DQ5-DR1 and DQ5-DR10 in the case of rheumatoid arthritis, predispose to impaired T-cell-mediated immune regulation. The haplotypes associated with impaired regulation are the combination of certain class II alleles and a yet unknown 'amplifier'. In this model, products of the HLA class II region are not involved in the presentation of particular organ-specific autoantigens. Therefore, HLA does not predispose to autoimmune disease per se, but rather fails to provide efficient protection. PMID- 11035124 TI - The effects of pulse methylprednisolone on matrix metalloproteinase and tissue inhibitor of metalloproteinase-1 expression in rheumatoid arthritis. AB - OBJECTIVE: To investigate the effects of a 1000 mg i.v. pulse of methylprednisolone succinate (pulse therapy) on the expression of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases-1 (TIMP-1) in the synovial membrane of the knee in patients with rheumatoid arthritis (RA). METHODS: Sequential arthroscopic biopsies of the knee were taken before and 24 h after pulse therapy (11 patients), at disease relapse (three patients) and after retreatment with pulse therapy (one patient). Immunoperoxidase staining for MMP-1 (interstitial collagenase), MMP-3 (stromelysin-1) and TIMP-1 was performed and the immunoreactive staining quantified by colour video image analysis. RESULTS: In the synovial lining layer, MMP-1 and TIMP-1 immunostaining was reduced by a mean of 47% (P = 0.02) and 72% (P = 0.05), respectively, 24 h after pulse methylprednisolone therapy. In the synovial sublining layer, MMP-1 was reduced by a mean of 51% (P = 0.08) and TIMP-1 by a mean of 73% (P = 0.02) 24 h after pulse methylprednisolone therapy. There was no change in MMP-3 staining in the synovial lining or sublining layer. CONCLUSIONS: High-dose pulse methylprednisolone therapy is associated with a rapid (within 24 h) and substantial decrease in the expression of MMP-1 and TIMP-1 but not MMP-3 in the synovial membrane in RA. PMID- 11035125 TI - Involvement of multinucleated giant cells synthesizing cathepsin K in calcified tendinitis of the rotator cuff tendons. AB - OBJECTIVES: Calcified tendinitis of the shoulder joint is a common painful condition. Resorption of the calcium deposits is one of the key events in the pathogenesis of this disease. The aim of this study was to examine whether the multinucleated giant cells that appear in this condition have osteoclast phenotypes. METHODS: Immunohistochemical and RNA in situ hybridization analysis of cathepsin K, a marker for osteoclasts, was performed in human surgical samples. RESULTS: The multinucleated cells located near the calcium deposits were positive for cathepsin K protein and mRNA. Reverse transcription-polymerase chain reaction using human cathepsin K-specific oligonucleotide primers confirmed that synthesis of cathepsin K mRNA occurs in the tissues of calcified rotator cuffs. CONCLUSION: The multinucleated giant cells which appear in the resorption area of calcium deposits in calcified tendinitis have the osteoclast phenotype. PMID- 11035126 TI - Elevation of plasma interleukin-18 concentration is correlated with disease activity in systemic lupus erythematosus. AB - BACKGROUND: Previous studies have indicated that the autoimmune phenomenon might be caused by an imbalance of T-helper cell (Th) cytokines. METHODS: We investigated the plasma concentrations of a novel pro-inflammatory Th1 cytokine, interleukin (IL)-18, and its inducer, IL-12, in patients with systemic lupus erythematosus (SLE) and correlated them with the SLE disease activity index (SLEDAI). Plasma IL-18 and IL-12 concentrations of 40 SLE patients and 18 sex- and age-matched control subjects were measured by enzyme-linked immunosorbent assay. RESULTS: Plasma IL-18 and IL-12 concentrations were significantly higher in SLE patients than in control subjects [median (interquartile range): IL-18, 320.0 pg/ml (164.4-475.6 pg/ml) vs 130.1 pg/ml (57.8-202.4 pg/ml), P<0.001; IL 12, 143.3 pg/ml (39.4-247.2 pg/ml) vs. 84.7 pg/ml (29.3-140.1 pg/ml), P<0.001]. Increases in IL-18 concentration correlated positively and significantly with SLEDAI score (r = 0.449, P = 0.004). CONCLUSION: The novel cytokine IL-18 might play a crucial role in triggering the inflammatory processes in SLE. PMID- 11035127 TI - Association of rheumatoid factors and anti-filaggrin antibodies with severity of erosions in rheumatoid arthritis. AB - OBJECTIVES: To evaluate and to compare the association of two types of autoantibodies-rheumatoid factors (RF) and anti-filaggrin antibodies (AFA)-with clinical severity and joint damage progression in rheumatoid arthritis (RA) patients. METHODS: In a cross-sectional study, we determined RF and AFA titres in 199 RA patients and 65 controls. Erosions apparent on X-rays were quantified using the Larsen score in 143 patients, and the distribution of these scores was studied according to disease duration in patients who were positive and negative for RF and AFA. RESULTS: RF were detected in 72% and AFA in 47% of RA patients. AFA were highly specific for RA (100%). RF positivity was correlated with the presence of subcutaneous nodules, sicca syndrome and the severity of erosions for a given disease duration. AFA positivity was correlated only with the presence of the HLA-DRB1 shared epitope. CONCLUSIONS: Since no significant correlation was observed between joint damage progression and AFA positivity, the determination of AFA does not appear to be useful in assessing the prognosis of RA. However, AFA, which appear early in RA, could be helpful for the diagnosis of RA in patients who do not fulfil four American College of Rheumatology criteria. PMID- 11035128 TI - Anticardiolipin antibody levels predict flares and relapses in patients with giant-cell (temporal) arteritis. A longitudinal study of 58 biopsy-proven cases. AB - OBJECTIVE: To evaluate the usefulness of anticardiolipin antibodies (aCL) in identifying flares and relapses in giant-cell arteritis. METHODS: We studied 58 consecutive patients with biopsy-proven temporal giant-cell arteritis. C-reactive protein and aCL serum levels were measured simultaneously at the time of diagnosis and at each out-patient visit until recovery. All observed episodes of a rise in C-reactive protein attributable to a precise cause, for which the simultaneous measurement of aCL was available, were analysed. RESULTS: The mean duration of clinical observation and serum aCL assessment was 34+/-18 and 24+/-11 months, respectively. Anticardiolipin antibody positivity (IgG or total antibodies > or =20 U) before treatment was found before treatment in 27 cases (46.6%) (mean 45.6+/-26 U/l, range 20-110 U). Levels of aCL decreased below 10 U with appropriate treatment in all patients except one, after a variable delay. No rise in aCL levels was recorded subsequently in any patient whose disease was controlled permanently. A significant rise in aCL was recorded in 20 of 27 (74%) of the flares or relapses of giant-cell arteritis, including seven of 12 flares in seven patients whose initial aCL level was <20 U vs none of the 28 inflammatory episodes unrelated to giant-cell arteritis (P<0.0000001). IgM aCL, infrequently found at diagnosis, was not associated with signs of disease activity. CONCLUSION: Serum aCL levels are useful in the detection of flares and relapses in giant-cell arteritis, with fairly good sensitivity (74%) and a specificity of 100%, and can be of value in distinguishing subclinical flares from infection. PMID- 11035131 TI - Circulating collagen metabolites in systemic sclerosis. Differences between limited and diffuse form and relationship with pulmonary involvement. AB - OBJECTIVE: To study collagen metabolites in systemic sclerosis (SSc) and their relationship with clinical manifestations of the disease. METHODS: Forty-eight SSc patients, 13 with a diffuse form (dcSSc), 23 with a limited form (lcSSc) and 12 with suspected SSc not fulfilling the ACR criteria, and 31 healthy controls were examined. Serum concentrations of aminoterminal type III procollagen peptide (PIIINP), aminoterminal and carboxyterminal type I procollagen peptides (PINP and PICP) and cross-linked carboxyterminal telopeptide of collagen I (ICTP) were determined by radioimmunoassay. RESULTS: Increased serum concentrations of ICTP were found in SSc patients compared with controls. Distinctly higher levels of ICTP were observed in dcSSc than in lcSSc. High serum ICTP was correlated with skin score and acute phase reactants, and with reduced pulmonary function. Serum PIIINP concentration was elevated in both lcSSc and dcSSc. CONCLUSION: Augmented collagen catabolism accompanies the increased collagen synthesis in SSc. Serum ICTP concentration is a marker of this feature and also reflects clinical severity. PMID- 11035130 TI - Do patients with rheumatoid arthritis established on methotrexate and folic acid 5 mg daily need to continue folic acid supplements long term? AB - BACKGROUND: It is postulated that some aspects of methotrexate toxicity may be related to its action as an anti-folate. Folic acid (FA) is often given as an adjunct to methotrexate therapy, but there is no conclusive proof that it decreases the toxicity of methotrexate and there is a theoretical risk that it may decrease the efficacy of methotrexate. OBJECTIVES: To look at the effect of stopping FA supplementation in UK rheumatoid arthritis (RA) patients established on methotrexate <20 mg weekly and FA 5 mg daily, to report all toxicity (including absolute changes in haematological and liver enzyme indices) and to report changes in the efficacy of methotrexate. METHODS: In a prospective, randomized, double-blind, placebo-controlled study, 75 patients who were established on methotrexate <20 mg weekly and FA 5 mg daily were asked to stop their FA and were randomized to one of two groups: placebo or FA 5 mg daily. Patients were evaluated for treatment toxicity and efficacy before entry and then at intervals of 3 months for 1 yr. RESULTS: Overall, 25 (33%) patients concluded the study early, eight (21%) in the group remaining on FA and 17 (46%) in the placebo group (P = 0.02). Two patients in the placebo group discontinued because of neutropenia. At 9 months there was an increased incidence of nausea in the placebo group (45 vs. 7%, P = 0.001). The placebo group had significantly lower disease activity on a few of the variables measured, but these were probably not of clinical significance. CONCLUSIONS: It is important to continue FA supplementation over the long term in patients on methotrexate and FA in order to prevent them discontinuing treatment because of mouth ulcers or nausea and vomiting. Our data suggest that FA supplementation is also helpful in preventing neutropenia, with very little loss of efficacy of methotrexate. PMID- 11035129 TI - The long-term effects of non-steroidal anti-inflammatory drugs in osteoarthritis of the knee: a randomized placebo-controlled trial. AB - BACKGROUND: Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to treat osteoarthritis (OA), though their long-term efficacy is uncertain. We report a comparison of the symptomatic responses to therapy with tiaprofenic acid, indomethacin and placebo over 5 yr. METHODS: A parallel-group, randomized, single-blind trial of patients with knee OA recruited 812 patients from 20 centres; 307 patients received tiaprofenic acid (300 mg b.d.), 202 indomethacin (25 mg t.d.s.) and 303 matching placebo for up to 5 yr. At the end of the parallel-group study, patients receiving tiaprofenic acid or placebo entered a 4 week blinded cross-over study of tiaprofenic acid or placebo, both given for 2 weeks. Assessments were at baseline, 4 weeks, then at 6-month intervals for up to 5 yr in the parallel group study and at 2-week intervals in the cross-over study. They comprised pain scores, duration of morning stiffness, patients' global assessments, paracetamol consumption, adverse reactions, withdrawals and functional outcomes. RESULTS: There were significant falls in overall pain scores in patients receiving NSAIDs compared with placebo at 4 weeks in the parallel group phase. Thereafter there were no advantages favouring active therapy. In the cross-over phase, pain scores were significantly lower in patients receiving tiaprofenic acid than placebo. Patients who had been receiving long-term tiaprofenic acid showed significant rises in their pain scores when receiving placebo therapy and vice versa. Adverse events were reported by 61% of patients receiving tiaprofenic acid, 63% on indomethacin and 51% on placebo. Potentially severe side-effects were rare; for example, there were only three cases of gastrointestinal bleeding on NSAIDs. The pattern of withdrawal was similar in patients taking NSAIDs and placebo in the parallel-group study; at 48 weeks 53% of the patients remained on tiaprofenic acid, 50% on indomethacin and 54% on placebo. CONCLUSIONS: NSAIDs significantly reduce overall pain over 4 weeks. This short-term responsiveness is retained, and even after several years of therapy with tiaprofenic acid pain scores increased over 2 weeks when it was changed to placebo. Our results do not show long-term benefits from the use of NSAIDs in OA and the majority of patients had persisting pain and disability despite therapy. PMID- 11035132 TI - Identification of candidate endothelial cell autoantigens in systemic lupus erythematosus using a molecular cloning strategy: a role for ribosomal P protein P0 as an endothelial cell autoantigen. AB - OBJECTIVE: To attempt to characterize the diversity and nature of antigens recognized by anti-endothelial cell antibodies (AECA) in patients with systemic lupus erythematosus (SLE) using a molecular cloning strategy. METHODS: AECA in sera of 15 SLE patients were measured by ELISA and Western blot analysis was used to examine the diversity of autoantigen targets in two clinically active patients. A human umbilical vein endothelial cell cDNA expression library was immunoscreened with sera from these two patients to identify their autoantigen targets. An anti-ribosomal P peptide antibody ELISA was used to assess the clinical significance of anti-ribosomal P protein antibodies in the sera of one patient. RESULTS: Significantly higher AECA levels were found in five patients with active disease and nephritis than in five patients with clinically inactive disease. Sera from two clinically active patients were found to recognize distinct spectra of autoantigens. The candidate autoantigens that were identified included (1) endothelial cell-specific plasminogen activator inhibitor; (2) the classical lupus antigen, i.e. ribosomal P protein P0; and (3) proteins never before described as putative autoantigens in SLE, including ribosomal protein L6, elongation factor 1alpha, adenyl cyclase-associated protein, DNA replication licensing factor, profilin II and the novel proteins HEAPLA 1 and HEAPLA 2 (human endothelial associated putative lupus autoantigens 1 and 2). In one patient, antibodies against ribosomal P protein P0 were predominant and levels of these antibodies correlated with total AECA levels, anti-DNA antibody titres, overall clinical score and renal disease in a longitudinal study. CONCLUSIONS: A panel of candidate endothelial autoantigens in SLE, which includes previously described autoantigens and novel targets, has been identified by a molecular cloning strategy. This novel molecular approach could also be applied to the identification of autoantigens in other autoimmune vascular diseases. PMID- 11035133 TI - 31P magnetic resonance spectroscopy in fibromyalgic muscle. AB - OBJECTIVE: To measure inorganic phosphate (Pi), phosphocreatine (PCr), ATP and phosphodiesters (PDE) in fibromyalgic muscle tissue by (31)P magnetic resonance spectroscopy. METHODS: A 1.5 Tesla scanner with a P 100 surface coil was used to examine 15 patients (mean age 49.9+/-14.3 yr) with fibromyalgia, according to the American College of Rheumatology criteria, and 17 healthy controls (mean age 30.2+/-5.8 yr). RESULTS: Compared with the controls, there were increases in the levels of PDE (+22%, P = 0.032) and Pi (+19%, P = 0.019) in the spectra of fibromyalgia patients, but there was no difference in pH. CONCLUSION: The metabolic differences we found may have been related to weakness and fatigue in the fibromyalgia patients, but they do not fully explain the fibromyalgia symptoms. PMID- 11035134 TI - IL-4 VNTR gene polymorphism in chronic polyarthritis. The rare allele is associated with protection against destruction. AB - OBJECTIVE: To evaluate the occurrence of variants of the interleukin 4 (IL-4) and IL-4 receptor (IL-4R) genes in patients with rheumatoid arthritis (RA) and their possible contribution to joint destruction. METHODS: Allelic frequencies for polymorphisms in the IL-4 [variable number of tandem repeat (VNTR) polymorphism in intron 3] and IL-4 receptor alpha chain (transition at nucleotide 1902) genes were assessed in 335 RA patients and 104 controls. Clinical indices of disease activity, disability and joint destruction and plasma levels of IL-1beta, IL-1Ra and sCD23 were assessed to evaluate a possible functional effect. RESULTS: Carriage of the rare IL-4(2) allele was higher in patients with non-destructive RA (40%) than in those with destructive RA (22.3%; odds ratio = 1.9, 95% confidence interval 1. 1-3.5, P = 0.0006) and in controls (26%, P = 0.002). Patients positive for this rare allele had significantly less joint destruction, assessed by the Larsen wrist index (P = 0.004) and a lower erythrocyte sedimentation rate (P = 0.04). A significantly higher carriage rate of IL-4(2) was seen in HLA-DR4/DR1(-) patients with non-destructive RA than in those with destructive RA. The IL-4 receptor polymorphism was not over-represented. Plasma levels of IL-1beta, IL-1Ra and sCD23, known to be modified by IL-4, were not different in individuals having different alleles. CONCLUSION: This IL-4 VNTR gene polymorphism may be a protective factor for severe joint destruction in RA that could be used as a prognostic marker early in the course of the disease. PMID- 11035135 TI - Effect of SR 49059, a V1a vasopressin receptor antagonist, in Raynaud's phenomenon. AB - OBJECTIVE: To assess whether vasopressin V1a receptor blockade reduces the abnormal vasoactive response to cold in patients suffering from Raynaud's phenomenon (RP). METHODS: SR 49059, an orally active, non-peptidic vasopressin V1a receptor antagonist, was given orally (300 mg once daily) to 20 patients with RP in a single-centre, double-blind, placebo-controlled, randomized cross-over study with two 7-day periods of treatment separated by 21 days of washout. Bilateral finger systolic blood pressure and skin temperature were assessed before and after immersion of the hand in cold water for 3 min (15 degrees C) during the screening phase and three times (before and 2 and 4 h after drug intake) on days 1 and 7 of each of the two treatment periods. Recovery of digital pressure and skin temperature was measured 0, 10, 20 and 32 min after the end of the cold immersion test. RESULTS: SR 49059 significantly attenuated the cold induced fall in systolic pressure by 14.5% (95% confidence interval 0-29; P = 0.045) on the most affected hand on day 7 compared with placebo. Temperature recovery after the end of the cold test showed a trend to enhancement 2 and 4 h after SR 49059 on day 7 (P = 0.060 and P = 0.062 respectively). The beneficial effects on finger pressure and temperature recovery were obtained without changes in supine blood pressure or in heart rate. CONCLUSION: SR 49059 given orally once a day for 7 days to patients with RP showed favourable effects compared with placebo on finger systolic pressure and temperature recovery after cold immersion, without inducing side-effects. PMID- 11035136 TI - Pharmacokinetic, pharmacodynamic and clinical effects of a humanized IgG1 anti CD4 monoclonal antibody in the peripheral blood and synovial fluid of rheumatoid arthritis patients. AB - BACKGROUND: CD4(+) T cells are important mediators in the pathogenesis of rheumatoid arthritis (RA). In this open-label, dose-escalating study, we examined the pharmacokinetic (PK), clinical, biological and immunological effects of a humanized IgG1 anti-CD4 monoclonal antibody (mAb), 4162W94, in the peripheral blood (PB) and synovial fluid (SF) of RA patients. METHOD: Twenty-four patients in four cohorts (six patients in each cohort) were allocated to be treated with five consecutive daily doses of 4162W94 (10, 30, 100 or 300 mg i.v.). Disease activity was measured by the American College of Rheumatology (ACR) criteria and disease activity score (DAS). We also measured 4162W94 concentration, the percentage of 4162W94-coated CD4(+) lymphocytes, percentage down-modulation of CD4, interleukin-6 (IL-6) and tumour necrosis factor alpha (TNFalpha) levels in the PB and SF. RESULTS: A direct relationship between 4162W94 dose, biological response and clinical outcome was seen. Treatment with 10 and 30 mg of 4162W94 for 5 consecutive days resulted in transient coating and down-modulation of CD4(+) lymphocytes, with little effect observed beyond the final dose. However, treatment with 100 and 300 mg resulted in sustained coating and/or down modulation for 3 weeks and 4 weeks, respectively, in PB and >4 weeks in SF in one patient from the 300 mg cohort. There was a dose-related moderate but transient depression in the CD4(+) lymphocyte count in most patients, with all but three returning to >0.40 x 10(9)/l or >75% baseline by the end of the study period. Significant clinical improvement (ACR 20%) was seen in only 1/6 patients in each of the 10- and 30-mg cohorts; however, 3/6 and 5/5 patients in the 100 and 300-mg cohorts, respectively, were ACR 20% responders. In addition, there were significant reductions in PB acute phase reactants as well as SF IL-6 and TNFalpha concentrations in parallel to clinical improvement. CONCLUSION: Data from this pilot study suggest that 4162W94 is a clinically active novel immunotherapeutic agent that may suppress inflammation in RA. PMID- 11035138 TI - Prevalence of articular hypermobility in schoolchildren: a one-district study in Barcelona. PMID- 11035137 TI - Is there an association of malignancy with systemic lupus erythematosus? An analysis of 276 patients under long-term review. AB - OBJECTIVE: To estimate the risk of malignancy in a UK cohort of patients with systemic lupus erythematosus (SLE) under long-term review. METHOD: The University College London Lupus Clinic Database was used to identify a cohort of 276 patients followed up prospectively between 1978 and 1999. Standardized incidence ratios and 95% confidence intervals for all cancers were calculated using age and sex-specific cancer incidence rates for the southeast of England. RESULTS: In total, 16 malignancies were diagnosed in 15 patients. However, five malignancies were diagnosed before the diagnosis of SLE and were therefore excluded from the final statistical analysis. One case of basal cell carcinoma was also identified, but this was also excluded from the final analysis as no comparable figures were available for the general population. Death as a direct consequence of the malignancy occurred in six (2.3%) patients, accounting for 22.6% of the deaths in our cohort of SLE patients. Compared with the general population, the overall estimated risk for all cancers was not increased in the lupus cohort (standardized incidence rate 1.16 (95% confidence interval 0.55-2. 13). Hodgkin's lymphoma was the only individual cancer that was increased in our cohort of patients [standardized incidence rate 17. 82 (95% confidence interval 0.45 99.23)]. CONCLUSIONS: In our cohort of patients with SLE we did not show an overall increased risk of malignancy. However, SLE was associated with an increased risk of Hodgkin's lymphoma compared with the general population. From our cohort of 276 patients, none of those treated with cyclosporin (3%) developed malignancy, and out of 49 (18%) patients treated with cyclophosphamide only one patient developed malignancy. Out of the 10 patients in the final analysis who developed malignancy, six had treatment with prednisolone, four with azathioprine, five with hydroxychloroquine and only one with cyclophosphamide. No statistical difference in the above cytotoxic therapy was observed between those patients who developed malignancy and those who did not. PMID- 11035139 TI - Acupuncture and rheumatoid arthritis. PMID- 11035140 TI - Re: Bingham and Emery. Resistant rheumatoid arthritis clinics--a necessary development? PMID- 11035141 TI - Successful treatment of pure red cell aplasia associated with systemic lupus erythematosus with cyclosporin A. PMID- 11035142 TI - Fatal haemophagocytic syndrome in the course of dermatomyositis with anti-Mi2 antibodies. PMID- 11035143 TI - Successful treatment of arthralgia with tamoxifen citrate in a patient with pachydermoperiostosis. PMID- 11035144 TI - Clinical protocol for monitoring of targeted therapies in rheumatoid arthritis. PMID- 11035145 TI - Do sex hormones play a role in fibromyalgia? PMID- 11035146 TI - Resolution of digital necrosis following treatment of multiple myeloma. PMID- 11035148 TI - Robin goodfellow PMID- 11035147 TI - Acromial apophysitis. PMID- 11035153 TI - Corrigenda PMID- 11035149 TI - MANUAL OF RHEUMATOLOGY. Edited By P. K. Pispati, N. E. Borges and S. S. Uppal. Indian rheumatism association (Mumbai), 1999 PMID- 11035154 TI - Differential mutation of transgenic and endogenous loci in vivo. AB - Although chemicals usually induce very similar frequencies of mutations in transgenes and endogenous genes in vivo when given acutely, chronic exposure to N ethyl-N-nitrosourea (ENU) produced a more complex pattern in which the endogenous locus was spared many mutations. Here, we demonstrate that the effect is neither ENU-specific nor locus-specific, and thus, may be important in the extrapolations of risk assessment and in understanding mutational mechanisms. During chronic mutagen exposure, mutations at the transgene accumulate linearly with time, i.e. in direct proportion to the dose received. In contrast, mutations at the endogenous gene are much less frequent than those of the transgene early in the exposure period and the accumulation is not linear with time, but rather accelerates as the exposure continues. Previous comparisons involved the endogenous Dlb-1 locus and the lacI transgene from the Big BlueMouse in the small intestine. These experiments involved the Dlb-1 locus and the lacZ transgene from the MutaMouse in the small intestine and the hprt locus and the lacZ transgene in splenocytes. Comparisons were made in both tissues after acute and chronic exposures to ENU, the original mutagen, and in the small intestine after exposures to benzo(a)pyrene. All comparisons showed that during chronic exposures mutations at the transgene accumulate linearly with the increasing duration of exposure, whereas induced mutations of the endogenous gene initially accumulate at a slower rate. Thus, the difference in mutational response observed during low chronic treatment is not unique to a particular transgene, endogenous gene, tissue, or mutagen used, but may be a general phenomenon of such genes. PMID- 11035155 TI - DNA adduct formation in northern pike (Esox lucius) exposed to a mixture of benzo[a]pyrene, benzo[k]fluoranthene and 7H-dibenzo[c, g]carbazole: time-course and dose-response studies. AB - The time-course and dose dependent formation of DNA adducts in juvenile northern pike (Esox lucius) following a single exposure to a mixture of benzo[a]pyrene (BaP), benzo[k]fluoranthene (BkF) and 7H-dibenzo[c,g]carbazole (DBC) were investigated by use of the (32)P-postlabelling assay. A complex adduct pattern was detected in liver and intestine of exposed fish. For the time-course studies fish were exposed either by oral administration or by intraperitoneal (i.p.) injection. Following a single i.p. injection of the mixture (40micromole/kg body weight of each substance) significantly elevated DNA adduct levels were detected in the liver after 1 day. Adduct levels were higher in liver than in intestine, in which significant elevation were detected from day 3 to 12. Following exposure via food (80micromole/kg body weight of each substance), adduct levels were detected in both liver and intestine 1 day after exposure, and continued to increase until day 3 in liver and day 6 in intestine. Calculation of a binding index, which compensates for differences in dosage, resulted in much higher adduct formation (five times in liver and 22 times in intestine) following oral exposure. Pikes receiving single oral doses of 12.5, 50, 100 or 200micromole/kg body weight of each substance exhibited significantly higher adduct levels in both liver and intestine compared to controls. Hepatic adduct levels were also higher in fish given 100 and 200micromole/kg compared to 12.5micromole/kg. Results from this study show that DNA adducts are rapidly formed in juvenile northern pike following both i.p. injection and feeding of a mixture of BaP, BkF and DBC. A maximum level was reached within a few days, which then persisted at approximately the same level for at least 9-12 days. The results also shows that higher levels of adducts were obtained following oral administration compared to i.p. injection, particularly in the intestine. PMID- 11035156 TI - Induction of centrosome injury, multipolar spindles and multipolar division in cultured V79 cells exposed to dimethylarsinic acid: role for microtubules in centrosome dynamics. AB - Role for microtubules in the induction of multiple microtubule organizing centers (MTOCs) and multipolar spindles by dimethylarsinic acid (DMAA), a methylated derivative of inorganic arsenics, was investigated with respect to the effects of microtubule disruption and reorganization. DMAA induced multiple signals of gamma tubulin, a well-characterized component of MTOCs in the centrosome, in a manner specific to mitotic cells. The multiple signals of gamma-tubulin were co localized with multipolar spindles caused by DMAA. Disruption of microtubules by nocodazole (NOZ) suppressed the appearance of centrosome injury caused by DMAA while disorganization of actin microfilaments by cytochalasin D did not. Post treatment incubation of cells in which multiple signals of gamma-tubulin caused by DMAA had been coalesced to one or two dots by NOZ caused the reappearance of mitotic cells with multiple signals of gamma-tubulin, in conjunction with reorganization of the microtubules. These results suggest a role for microtubules in the dynamic behavior of the mitotic centrosome. DMAA induced aberrant cytokinesis, such as tripolar and quadripolar division, in a concentration dependent manner. These results, together with the findings of earlier studies, suggest that the centrosome is the primary target for the induction of multipolar spindles by DMAA and the resultant induction of multinucleation and multipolar division. PMID- 11035157 TI - Benzo(a)pyrene activates L1Md retrotransposon and inhibits DNA repair in vascular smooth muscle cells. AB - Benzo(a)pyrene (BaP) modulates vascular smooth muscle cells (vSMCs) from a quiescent to proliferative phenotype, a shift associated with activation of L1Md retrotransposon [K.P. Lu, K.S. Ramos, Biochem. Biophys. Res. Commun. 253 (1998) 828-833]. The present studies were conducted to evaluate L1Md activation profiles in murine vSMCs treated with BaP or its oxidative metabolites, and to screen for possible insertional mutations into p53 and retinoblastoma (RB) genes. We also sought to examine the profile of DNA damage and repair in BaP-treated vSMCs. Northern analysis revealed that BaP (0. 03-3microM), and its major reactive 7,8 diol metabolite (0. 03-3microM), activate L1Md gene in a concentration-dependent manner. Two other metabolites, 3-OH BaP and 3,6-BaP quinone (0.03-3microM), as well as hydrogen peroxide (25-75microM) also activated L1Md. No insertional mutations into either p53 or RB genes were observed in vSMCs treated with BaP in vitro, although a slight elevation of p53 mRNA was observed as early as 4h after chemical challenge. Treatment of vSMCs with 3 or 30microM BaP for 4h increased unscheduled DNA synthesis (UDS) 1.4- and 2.5-fold, respectively. Challenge with 0. 3microM BaP for 24h inhibited DNA repair capacity in vSMCs for up to 48h. These results demonstrate that BaP and its oxidative metabolites activate L1Md retrotransposon in vSMCs, which coupled to DNA damage and inhibition of DNA repair are part of the atherogenic response elicited by BaP and related hydrocarbons. PMID- 11035158 TI - Mutagenicity of nitrosamines in methyltransferase-deficient strains of Salmonella typhimurium coexpressing human cytochrome P450 2E1 and reductase. AB - Although dialkylnitrosamines are environmentally significant carcinogens, the use of short-term bioassays to assess the mutagenic potential of these compounds is problematic. The Ames test, a mutagenicity assay based on the reversion of Salmonella typhimurium histidine auxotrophs, is the most widely used bioassay in genetic toxicology, but the traditional Ames tester strains are largely insensitive to dialkylnitrosamine mutagenicity. We have constructed two mutagenicity tester strains that co-express full-length human cytochrome P450 2E1 and P450 reductase in S. typhimurium lacking ogt and ada methyltransferases (YG7104ER, ogt- and YG7108ER, ogt-, ada-). These new strains are susceptible to dialkylnitrosamine mutagenicity in the absence of an exogenous metabolic activating system (S9 fraction). Mutagenicity is dependent upon the coexpression of P450 2E1 with P450 reductase and is similar to or greater than that obtained with the parental strains in the presence of S9 fraction from ethanol-induced rat liver. These strains were also sensitive to nitrosamines with longer alkyl side chains including diethylnitrosamine, dipropylnitrosamine and dibutylnitrosamine. Mutagenicity decreased with alkyl chain length, consistent with the stringency of the ada-encoded enzyme for methyl and ethyl DNA adducts. These new strains may prove useful in the evaluation of nitrosamine contamination of food and environmental samples. PMID- 11035159 TI - Simultaneous measurement of the frequencies of intrachromosomal recombination and chromosome gain using the yeast DEL assay. AB - The yeast DEL assay measures the frequency of intrachromosomal recombination between two partially-deleted his3 alleles on chromosome XV. The his3Delta alleles share approximately 400bp of overlapping homology, and are separated by an intervening LEU2 sequence. Homologous recombination between the his3Delta alleles results in deletion of the intervening LEU2 sequence (DEL), and reversion to histidine prototrophy. In this study we have attempted to further extend the use of the yeast DEL assay to measure the frequency of chromosome XV gain events. Reversion to His(+)Leu(+) in the haploid yeast DEL tester strain RSY6 occurs upon non-disjunction of chromosome XV sister chromatids, coupled with a subsequent DEL event. Here we have tested the ability of the yeast DEL assay to accurately predict the aneugenic potential of the diversely-acting, known or suspected aneugens actinomycin D, benomyl, chloral hydrate, ethyl methanesulfonate (EMS), methyl methanesulfonate (MMS), and methotrexate. Actinomycin D and benomyl strongly induced aneuploidy. EMS and methotrexate modestly induced aneuploidy, while chloral hydrate and MMS failed to illicit any significant induction. In addition, by FACS-analysis of DNA content it was shown that the majority of both spontaneous- and chemically-induced His(+)Leu(+) revertants were heterodiploid. Thus, our results indicate endoreduplication of almost entire chromosome sets as a major mechanism of aneuploidy induction in haploid Saccharomyces cerevisiae. PMID- 11035160 TI - Radiosensitivity of HCV cells and their v-ras and v-raf transfectants. AB - In the present work, it was found that transfection of cultivated urothelial cells HCV-29 with v-raf and v-ras oncogenes increased their sensitivity to ionizing radiation, as documented by clonogenic studies. Flow cytometry study showed, that HCV-29 and their v-ras transfectants were arrested around middle S phase, whereas v-raf transfectants randomly at each point of S phase. This unusual reaction of HCV-29 v-raf cells may partially explain studies of P21(WAF1/CIP1) and GADD45 genes, whose transcripts were found only in these cells. Increased radiosensitivity of v-ras transfectants is probably associated with c-JUN protein overexpression. Altogether the obtained results suggested different mechanism of reaction on irradiation of v-raf and v-ras transfected cells. PMID- 11035161 TI - Mutagenic effect by phenylalanine during gamma-irradiation of plasmid DNA in aqueous solution under oxic conditions. AB - Irradiation of DNA in aqueous solution or in cells with gamma-rays results in different mutational spectra, indicating that in both situations different patterns of DNA damages are induced. One of the causes for these different types of damages might be the formation of secondary, organic radicals, if cells are irradiated. Some organic compounds, including the amino acid phenylalanine, are well known to produce radicals during irradiation. Under oxic conditions these secondary radicals react with oxygen, thus forming peroxyl radicals which can be very harmful to DNA, and which may, therefore, induce DNA damage leading to mutations. This study examines the influence of the presence of phenylalanine during gamma-irradiation of DNA in aqueous solution under oxic conditions. The results indicate that the formation of phenylalanine radicals influences the types of induced mutations in the gamma-radiation-induced mutation spectrum. The most prominent difference is the increase in G:C to T:A transversions and the decrease in G:C to A:T transitions in the presence of phenylalanine. Further, it appears that the gamma-radiation-induced mutation spectrum after irradiation of DNA in aqueous solution is more comparable to the intracellular gamma-radiation induced mutation spectrum in E. coli cells, if phenylalanine is present during irradiation. Therefore, these results suggest that the presence of phenylalanine during irradiation of DNA in aqueous solution gives a better impression of gamma radiation-induced mutations in bacterial systems than water only. PMID- 11035162 TI - Antimutagenic effects of wheat bran diet through modification of xenobiotic metabolising enzymes. AB - Diets containing wheat bran (WB) protect against cancers of the colon or breast in rats, and may be beneficial in humans. In a previous study of rats treated with the carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), inclusion of 10% wheat bran in the diet led to an apparent reduction in IQ metabolites but not of intact IQ in plasma. In the present study, male Wistar rats were fed diets containing 0, 10 or 20% wheat bran, and effects on xenobiotic metabolising enzymes compared. Wheat bran-supplementation showed differential effects on phase I enzymes, significantly increasing the activity of hepatic cytochrome P450 isozyme CYP3A2, but slightly reducing the activity of CYP1A1/2. The activities of both hepatic phase II detoxification enzymes glutathione-S-transferase and glucuronosyl transferase were also reduced. Western blotting revealed similar effects on expression of the proteins. Interestingly, the expression of xenobiotic metabolising enzymes (XME) in the colon appeared to be modulated independently of hepatic XME. Although the wheat bran-supplemented diet still led to an increased expression of CYP3A, it now slightly increased CYP1A in the colon. However, 20% wheat bran significantly increased the expression of both glutathione transferase isozymes, GST A1 & A2, in the colon. Natures Gold (NG) is a commercial wheat bran derivative which is lower than wheat bran in dietary fibre, but enriched in vitamins, minerals and various phytochemicals. Dietary supplementation with 20% Natures Gold led to similar trends as seen in wheat bran fed rats, but more potent effects in both hepatic and colonic enzymes. The significance of these changes for activation of carcinogens to mutagenic metabolites was investigated using the Salmonella/mammalian microsome mutagenicity test. The activation of IQ and benzo[a]pyrene, but not cyclophosphamide, to a mutagen by hepatic S9 from wheat bran-fed or Natures Gold fed rats was significantly reduced compared with S9 from animals on a diet lacking wheat bran. We suggest that modulation of xenobiotic metabolising enzymes may be an important component of cancer protection by wheat bran, and this effect may relate to micronutrients or cancer-protective non-nutrient phytochemicals rather more than to dietary fibre. PMID- 11035163 TI - Radiation-induced genomic instability: a paradigm-breaking phenomenon and its relevance to environmentally induced cancer. AB - The existing paradigm governing radiobiology which is fundamental to the estimation of environmental radiation risk, cannot explain the phenomena of radiation induced genomic instability and the bystander effect. Both effects can, however, be understood in terms of the dynamical genome concept, qualitatively described herein. The dynamical genome concept may find further application in better understanding other aspects of biology, most notably the cancer process in general and the consequences of genetic modification. PMID- 11035164 TI - Erratum to "Accumulation of DNA damage in the organs of mice deficient in gamma glutamyltranspeptidase" PMID- 11035165 TI - A contribution to the knowledge of the pteridosperm genera Pseudomariopteris Danze-Corsin nov. emend. and Helenopteris nov. gen. AB - The macromorphology and epidermal anatomy of three Late Carboniferous-Early Permian pteridosperm species, conventionally assigned to the form-genus Pseudomariopteris Danze-Corsin, are described from the Stephanian of the Blanzy Montceau Basin (Central France). The generic diagnosis of Pseudomariopteris is emended, and P. busquetii is designated as the type species of the genus. The combination P. cordato-ovata (Weiss) Gillespie et al. is validated to replace the illegitimate name P. ribeyronii (Zeiller) Danze-Corsin. The species diagnoses of P. busquetii and P. cordato-ovata are emended. On the basis of its epidermal anatomy, P. paleaui, another species orininally described from Central France, is transferred to the newly established genus Helenopteris Krings et Kerp, and the new combination Helenopteris paleaui is introduced. Lectotypes of P. busquetii and H. paleaui are selected. In addition, specimens from other basins are (re )examined and illustrated. Seeds attached to a P. busquetii specimen suggest a callistophytalean relationship for Pseudomariopteris. Compression fossils and cuticles of Pseudomariopteris and Helenopteris are interpreted in terms of palaeoecology: Pseudomariopteris busquetii and P. cordato-ovata had climbing/scrambling growth habits; specialized climber hooks can be demonstrated for both species, either in compression or in cuticular preservation. Although the growth habit of H. paleaui is not yet entirely clear, the size of its fronds suggests that this taxon most probably was also a scrambler/climber. Finally, based on epidermal features, a tentative reconstruction of stages of the pinnule ontogeny of H. paleaui is provided. PMID- 11035166 TI - Spatial distribution of calcareous dinoflagellate cysts in surface sediments of the Atlantic Ocean between 13 degrees N and 36 degrees S. AB - To enhance the limited information available about the palaeo-ecological significance of calcareous dinoflagellates, we have studied their lateral distribution in surface sediments of the equatorial and south Atlantic between 13 degrees N and 36 degrees S. Calcareous dinoflagellate cysts appear to be widely distributed throughout the studied area. In the surface sediments, concentrations (cyst per gram dry sediment) of the vegetative stage Thoracosphaera heimii are generally higher than that of the (presumably) calcareous resting cysts. Distribution patterns in surface sediments of Orthopithonella granifera (Futterer) Keupp and Versteegh, Rhabdothorax spp. Kamptner., Sphaerodinella albatrosiana (Kamptner) Keupp and Versteegh S. albatrosiana praratabulated, Sphaerodinella tuberosa var. 1 (Kamptner) Keupp and Versteegh and S. tuberosa var. 2 and the ratios between these species have been compared with temperature, salinity, density and stratification gradients in the upper water column. Rhabdothorax spp. is characteristically present in sediments of more temperate regions characterized by high seasonality. Dinoflagellates producing these cysts are able to tolerate high nutrient concentrations, and mixing of the water column. S. albatrosiana is abundant in regions characterized by high sea surface temperatures and oligotrophic surface water conditions. In contrast, the distribution of S. tuberosa var. 2 is negatively related to temperature. The other cyst species did not show a characteristic pattern in relation to the studied environmental gradients.The ratio of Sphaerodinella tuberosa var. 2 to Orthopithonella granifera can be used for reconstructing the presence of stratification in the upper 50m of the water column, whereas the ratios of S. tuberosa var. 2 to Sphaerodinella albatrosiana and of O. granifera to Rhabdothorax spp. might be used for palaeotemperature reconstructions. Calcareous dinoflagellate cysts are abundant in oligotrophic areas and may be useful for the reconstruction of palaeoenvironmental conditions. PMID- 11035167 TI - Structurally preserved sphenophytes from the Triassic of Antarctica: reproductive remains of Spaciinodum. AB - Permineralized cones found organically attached to Spaciinodum collinsonii stems are described from the early Middle Triassic silicified flora from the Fremouw Formation of Antarctica, and the species diagnosis is emended to include the reproductive specimens. The apical cones are organized into internodal and leaf bearing nodal regions. Nodal septations span the central pith and cortex, and thin fimbrils subdivide the internodal areas into smaller chambers. The vascular system consists of 31-33 continuous bundles that do not alternate in position between successive nodes and internodes. Simple sporangia are associated with the cortical chambers and occur in one whorl on the axis. Spores are small, lack elaters, and have no discernible ultrastructure preserved, and they are interpreted to be immature. The Antarctic cones are different in structure from typical cones of modern and fossil members of Equisetales; however, they share similarities with some morphologically aberrant cones of extant Equisetum and several Late Paleozoic and Mesozoic compression-impression fossils. Spaciinodum is now the most complete anatomically described Mesozoic sphenophyte. PMID- 11035168 TI - Re-assignment of the Affinities of the Fossil Pollen Type Tricolpites trioblatus Mildenhall and Pocknall to Wilsonia (Convolvulaceae) and a reassessment of the ecological interpretations. AB - Tricolpites trioblatus Mildenhall and Pocknall was described from Upper Miocene Pliocene sediments of New Zealand and attributed to the Hebe complex (Scrophulariaceae), which is common in the New Zealand vegetation, especially in montane and subalpine habitats. Pollen in Miocene-Pliocene sediments in central Australia is identified with T. trioblatus, and the depositional situations included shallow lakes, with fresh or brackish waters, sometimes becoming saline. The affinities of T. trioblatus are re-examined in the light of these disparate environments in Australia and New Zealand. It has been found that all the fossil grains examined are more comparable to pollen of Wilsonia, and perhaps Cressa (Convolvulaceae), than to those of the Hebe complex. Wilsonia and Cressa are found in salt marshes, hence affinities with them are ecologically more credible for central Australia. T. trioblatus is found in late Eocene sediments deposited under episodic marine transgressions; an environment likely to stimulate the evolution of new species tolerant to saline conditions. PMID- 11035169 TI - Pollen input to, and incorporation in, two crater lakes in tropical northeast Australia. AB - Pollen input to the water surfaces of maar crater lakes Barrine and Eacham (1km(2) and 0.5km(2) respectively, ca. 720m a.s.l.), surrounded by rainforest in northeastern Australia, were measured in floating Tauber traps for a variety of periods spanning the years 1978 to 1985. The mean weekly total pollen catch is 75% of that estimated for the surrounding region of varied vegetation. Regardless of their sources, the influxes of pollen taxa are unevenly distributed over the lakes' surfaces. There is similar variation through time, with some indication that this might be smoothed over periods of 5years or more. There are no inflow streams and pollen input down surrounding slopes is negligible. Mean total pollen influx to seston traps close to the lakes' bottoms (1979 to 1988) is <5% of that to the Tauber traps, a discrepancy attributed to trap design. In contrast, mean annual influx measured by the Tauber traps is similar to that estimated from dated sediment samples younger than 1966 AD at Lake Barrine. The proportions of different pollen taxa are less affected by trap position or period of exposure than are their influxes; their mean percentages for Tauber traps, seston traps and sediment samples are all rather similar.The similarity between the mean contribution of exclusively rainforest pollen to the lakes' surfaces (56%) and to Oldfield traps in rainforest interiors (51%) emphasizes the importance of the rainforests, which clothe the crater walls, as sources of pollen to the lakes. The impact of pollen from other vegetation in the region, although present, is much smaller than at interfaces between rainforest and other vegetation types. All this pollen is mixed within and above the crater and delivered to the lakes' surfaces by gravity and frequent rainfall. Limnological processes redistribute it in the water body before it is incorporated into the deep-lake sediments.The value and limitations of current pollen transport and accumulation theory are noted in relation to sites with morphologies similar to these two crater lakes and to the irregular flowering and floristic inhomogeneity of tropical rainforest. The potential for the use of modern pollen input data in the interpretation of pollen analyses from the sediments of such sites is explored. PMID- 11035170 TI - Leaves of Cercocarpus mixteca n. sp. (Rosaceae) from Oligocene sediments, near Tepexi de Rodriguez, Puebla. AB - A new species of Cercocarpus, Cercocarpus mixteca Velasco de Leon & Cevallos Ferriz, is described based on leaf impressions from the Los Ahuehuetes locality, near Tepexi de Rodriguez, Puebla, Mexico. The lamina is obovate, 1.3cm in length by 0.5cm in width, has a serrate margin in its distal fourth, craspedodromous venation with a single straight mid-vein and two to four pairs of secondary ones, and areols that tend to be quadrangular in shape. A phenetic analysis of the agglomerative, non-hierarchical type, with mean linkage, is applied using 22 OTUs and 34 character states. The morphological characters observed on the leaves of the new fossil plants support the recognition of a new taxon closely related to the extant Cercocarpus paucidentatus growing naturally in northern Mexico. Its microphyll size corresponds with the temperate to xeric climate postulated for the Los Ahuehuetes locality; this further suggests that some taxa, like Cercocarpus, have a long history in low latitude North America. In this particular case, the extant Cercocarpus fothergilloides and Cercocarpus macrophyllus could, as they were able to colonise new humid and xeric areas, represent descendants of C. mixteca. PMID- 11035171 TI - Late Holocene history of savanna gallery forest from Carimagua area, Colombia. AB - The pollen record of a 65cm long core Laguna Carimagua-Bosque (4 degrees 04'N, 70 degrees 13'W) shows the late Holocene environmental history from a lake located within the gallery forest of the savannas of the Llanos Orientales of Colombia. Nine AMS radiocarbon dates of the organic deposits show that the core represents the period from ca. 1300(14)CyrBP to the present. The lake evolved from an active drainage system.During the period from ca. 1300 to 875(14)CyrBP (zone CMB-Ia), Mauritia-dominated swamp and gallery forest was present, dominated by Cecropia, and later also Acalypha and Alchornea. From 875 to 700(14)CyrBP (zone CMB-Ib), the lake was completely surrounded by gallery forest. Mauritiella and Cecropia occurred around the lake. Cecropia pioneer forest reached its greatest abundance and became gradually replaced by a more species-rich gallery forest, including Acalypha, Alchornea, Euterpe/Geonoma, Moraceae/Urticaceae, Piperaceae, and Virola. From 700 to 125(14)CyrBP (zone CMB-II), Cecropia lost its dominant role, and Mauritiella palms became more frequent. The main vegetation categories were swamp forest, gallery forest, understory elements, savanna shrubs and trees, and grass savanna. From 125(14)CyrBP to recent (zone CMB-III), the plant diversity in the gallery forest became highest, Mauritiella became very abundant, and among the savanna elements, woody Didymopanax increased.Comparison of four pollen records from savanna sites shows that pollen of savanna vegetation is markedly underrepresented in lake sediments when the lake lies within the gallery forest. As most of the drainage system of a savanna is hidden by gallery forest, we also expect a significant underrepresentation of the savanna ecosystem in river transported pollen assemblages. PMID- 11035172 TI - Hearing impairment prevention in developing countries: making things happen. AB - It is estimated that at least two thirds of the world's population of persons with disabling hearing impairment reside in developing countries. Yet, little and slow progress have been reported in these countries towards tackling this problem principally on account of inadequate resources. The prospects for improvement remain uncertain. This paper re-examines the peculiar nature of hearing impairment prevention within the context of the existing health-care needs of most of these nations. It establishes that the failure to recognize the dynamics of the social change that underlie an effective national programme on hearing impairment prevention may, in itself, forestall a successful and sustainable outcome even when more resources become available. PMID- 11035173 TI - The hearing profile of Nigerian school children. AB - The paucity of up-to-date and representative epidemiological data on hearing disorders in Nigeria has been observed as undermining the effective advocacy of prevention initiatives. This study attempts to address this problem by evaluating the prevalence and pattern of hearing impairment in school entrants. Parental interviews, otoscopy, pure-tone audiometric screening (frequency 0.5-4 kHz) and tympanometric examinations were conducted for a representative sample of 359 school children in an inner city area of Lagos. The prevalence of hearing loss was 13.9%. Middle ear abnormalities were noted in 20.9% of the study population, of which 18.7% were reported with otitis media with effusion. Impacted cerumen, documented in 189 children (52.6%), was the most common disorder. It showed significant association with hearing loss (P<0.001) and school performance (P<0.01). Tympanic membrane abnormalities were observed in 144 (40.1%) children. Of these, 45 (31.3%) showed abnormal tympanograms while 28 (19.4%) reported hearing loss (P<0.05). The early detection and management of hearing problems is relatively rare, thus precluding the determination of possible aetiological factors for the observed abnormalities. Poor public awareness, dearth of relevant facilities and the lack of early screening programmes are major known contributory factors. The well established national immunisation programme offers a cost-effective platform within the primary health-care system for addressing the high prevalence of hearing abnormalities in school children. PMID- 11035174 TI - The application of computer-enhanced imaging to improve preoperative counselling and informed consent in children considering bone anchored auricular prosthesis surgery. AB - OBJECTIVES: Bone anchored auricular prostheses have become a valuable option in the treatment of congenital and acquired deformities of the pinna. However, preoperative counselling and informed consent remains a challenging issue. Until recently it has been difficult to provide the child with a realistic prediction of their own postoperative appearance. This is particularly relevant when a remnant pinna needs to be excised prior to the second stage. The potential for psychological repercussions and the possibility that remnant excision might compromise future autologous tissue reconstruction make it imperative that the decision to proceed with surgery is founded on the best possible information. METHODS: The authors describe the use of computer enhanced images using the Adobe Photoshop (Apple Mac. Inc.) software package to provide such a preview. This technique is used in the outpatient clinic as an adjunct to counselling provided by clinic staff and is reinforced by meeting children who have already enrolled on to the implant programme. Children are encouraged to follow the stages of their planned operation on the computer screen, providing an accurate insight into the physical consequences of surgery. RESULTS: Our experiences suggest that this approach has encouraged a better qualitative understanding of implant surgery which has helped to foster the on-going commitment that is required to maintain a long-lasting, trouble-free implant site. CONCLUSIONS: This application of the Adobe Photoshop package has strengthened our basis for a personal informed consent and has provided an opportunity to lessen the adverse psychological consequences of such irreversible surgery. It is commended for its simplicity as it employs established software to enhance photographic prints or slides taken from the child's clinical records. PMID- 11035175 TI - Gastroesophageal reflux, allergy and chronic tubotympanal disorders in children. AB - The purpose of this study is to examine the relation between gastroesophageal reflux and allergy as possible causes of chronic tubotympanal pathology. The 30 examined children (ages 2-13) were divided into two groups based on the otological criteria. The 16 examined children suffered from a secretory otitis, which lasted more than four months. Upon further examination with a microscope, seven of these children exhibited symptoms of the adhesive process of the middle ear. Furthermore, 14 patients suffered from a recurrent otitis, i.e. more than five cases of otitis per year, while five patients from this group suffered from a chronic otitis with a central defect of the tympanum. The method used for the examination of the gastroesophageal reflux consisted of a continual 24 h esophageal pH monitoring. The particular apparatus used for this included antimony electrode (Synetics Medical, Sweden), while the analysis we performed was processed through the PC software program Gastrosoft Inc. The reflux index higher than five was considered pathological. At the time of the gastroesophageal reflux examination, we also performed the allergological analysis. The presence of allergy was confirmed by three methods: the positive allergological anamnesis, the positive skin pick test and by the elevated quantities of specific IgEs (Pharmacia CAP system). The examination resulted in the following: 18 of the examined children suffered from the pathological gastroesophageal reflux (60%); further seven of our patients tested positive on the allergological test (23%); and the four who tested positive for allergy also suffered from the pathological gastroesophageal reflux (13%). In comparison with allergies, the pathological GER was substantially more frequent in the patients who suffered from chronic tubotympanal disorders. PMID- 11035176 TI - Smell and taste in children with simple obesity. AB - In 30 children, aged 10-16 years and suffering from simple obesity, significantly lowered odour detection thresholds were noted. The thresholds were lower than the average for a given age group in around 20% of obese children in cases of odours stimulating olfactory nerve and in around 57% in cases of substances stimulating olfactory and trigeminal nerves. Odour identification thresholds were similarly affected, with identification of olfactory nerve plus trigeminal nerve stimulating odours affected more than twice as frequently. In 77% cases also the electrogustometric thresholds were found below the normal range values when anode was used as the stimulating electrode. The detected alterations may be linked to metabolic disturbances, which accompany simple obesity. PMID- 11035177 TI - Alteration of clinical picture and treatment of pediatric acute otitis media over the past two decades. AB - The clinical picture of acute otitis media (AOM) has changed greatly over the last few decades: serious complications have almost disappeared but more and more children suffer from recurrent middle ear infections and prolonged silent effusion. In this retrospective study we registered all AOM attacks among children under 10 in two rural municipal areas of Finland during 12-month-periods in 1978-79 and 1994-95. In addition to epidemiological data, the clinical picture and given treatments were recorded. Between study periods the number of children with recurrent AOM attacks increased heavily. The percentage of spontaneous otorrhea decreased from 6.0 to 3.3% (P=0.01) and the proportion of afebrile patients increased from 64.0 to 73.4% (P=0.002). In 1978-79 there were significantly more cases of diagnostic symptoms lasting over 24 h. Prescribing penicillin-V as a primary treatment decreased from 80.2 to 10.5% in favor of broad-spectrum antibiotics. The primary treatment with penicillin-V was associated with a decreased risk of recurrences. Acute tympanocentesis was performed less often and the incidence of surgical treatments (adenoidectomy and/or tympanostomy tube insertion) doubled from 6.2 to 12.4% of the acute cases. Although the clinical picture of AOM has become milder, children are treated with wider spectrum antibiotics. Nevertheless, a large number of children suffer from sequelae and there is a greater need for surgical treatments than 20 years ago. PMID- 11035178 TI - Reversible cardio-pulmonary changes due to adeno-tonsilar hypertrophy. AB - Adeno-tonsillar hypertrophy, with signs of upper airway obstruction is a common presentation in ENT clinics. Recently it is identified as a major cause of sleep apnea syndrome. Several isolated case reports of pulmonary hypertension and corpulmonale appeared in the literature. The authors report two such children aged less than 2 years with cardio-pulmonary changes occurring secondary to chronic adeno-tonsillar hypertrophy that were successfully treated with the surgical removal. PMID- 11035179 TI - Hypernasality--a rare initial symptom of a cerebellar astrocytoma. AB - Nasality is a disorder due to nasal resonance, which may be induced by a variety of etiologies. Transitional hypernasality is frequently seen in children after adenoidectomy. The alleged post-surgical hypernasality in the case presented was shown to be related to the late detection of an astrocytoma of the cerebellum and the brain stem in a 6-year-old boy. This case was characterized by increased hypernasality which failed speech therapy. A developing one-sided vocal fold palsy in combination with an ipsilateral soft-palate palsy indicated further investigation. Computerized tomography (CT) and magnetic resonance imaging (MRI) revealed a brain stem-tumor with a maximum size of 6 cm involving parts of the cerebellum. These findings demonstrated the need for a strict follow-up, even after adenoidectomy, in the presence of hypernasality for identifying concurrent etiologies as well as cases suitable for speech therapy. PMID- 11035180 TI - Pediatric coin ingestion: an unusual presentation. AB - A 35-month-old child presented to the Emergency Department with a suspected coin ingestion. A physical examination and radiographic examination revealed no evidence of the coin, and the child was prepared for discharge. When the child continued to refuse to drink, digital examination of the hard palate revealed the coin lodged behind the upper incisors. It was only possible to visualize when the patient's neck was fully extended. This case represents an unusual presentation of coin ingestion. It points out the importance of a meticulous physical examination and the need for reevaluation when findings are contradictory. PMID- 11035181 TI - The pathology of the temporal bones of a child with acquired cytomegalovirus infection: studies by light microscopy, immunohistochemistry and polymerase-chain reaction. AB - STUDY DESIGN: The first case of an acquired cytomegalovirus (CMV) infection of the inner ear is reported in a 3-year-old girl in remission from acute lymphocytic leukemia. METHODS: Horizontal sections of the temporal bones were studied by light microscopy and immunohistological staining by avidin-biotin complex-technique was performed on selected archival sections. Three sections were processed for detection of the virus genome by the polymerase chain reaction (PCR). RESULTS: By light microscopy the epithelium of the endolymphatic sac, the utricle and the semicircular canals showed deeply stained acidophilic inclusions and the stria vascularis had a loose structure especially in the intermediate layer. The changes were limited to the non-sensory parts of the labyrinth and no CMV type cells were observed in the organ of Corti. There was a loss of inner and outer hair cells and loss of cochlear ganglion cells caused by either the virus or treatment with gentamicin. Standard immunohistochemistry failed to demonstrate staining with CMV antibodies, but PCR, demonstrated CMV-DNA in one section. CONCLUSION: Molecular techniques may be able to detect acquired CMV infections in archival pediatric bones temporal bones. The histologic findings in the labyrinth were milder, however showed some similarity to children with congenital CMV labyrinthitis. PMID- 11035182 TI - Continuous enantioselective esterification of trans-2-phenyl-1-cyclohexanol using a new Candida rugosa lipase in a packed bed bioreactor. AB - Enantioselective resolution of trans-2-phenyl-1-cyclohexanol (TPCH) by a Candida rugosa lipase, obtained by fermentation in the laboratory, and immobilised on EP100 polypropylene powder has been carried out using isooctane as solvent and propionic acid as esterifying agent. The study have included the utilisation of this biocatalyst in a batch process and the optimisation of the esterification conditions by means of a Box-Hunter-based experimental design. The main variables controlling the process, concentration of acid and alcohol, have been numerically optimised using initial esterification rate as objective function. The optimal concentrations for the batch process were 50 mM for the alcohol and 71 mM for the acid. This esterification reaction kinetics corresponded to a reversible Michaelis-Menten kinetic law for the optimal conditions, which has permitted to select a plug-flow packed bed bioreactor as the most appropriate configuration to minimise the residence time and to avoid shear stress effect on the biocatalyst. The behaviour of the continuous packed bed bioreactor at two different residence times (302 and 582 min) was in accordance with predictions from batch experiments, with slightly deviations (less than 10%). Continuous experiments maintained high values of enantioselectivity (enantiomeric factor was practically 1) and conversion near equilibrium value (35%) when long-time operation was carried out. Besides, long-time stability of biocatalyst has permitted to scale up the production of enantioenriched (1R,2S)-TPCH propionate to yield gram quantities. PMID- 11035183 TI - Estimation of kinetic rates in batch Thiobacillus ferrooxidans cultures. AB - In this work, the key problem of estimation in bioprocesses when no structural model is available is dealt with. A nonlinear observer-based algorithm is developed in order to estimate kinetic rates in batch bioreactors. The algorithm uses the measurements of biomass concentration and either substrate concentration or redox potential to perform the estimation of the respective specific kinetic rates. For this purpose, a general mathematical model description of the process is provided. The estimation algorithm design is based on a nonlinear reduced order observer. The observer performance is validated with experimental results on a Thiobacillus ferrooxidans batch culture. PMID- 11035184 TI - Expression-independent consumption of substrates in cell-free expression system from Escherichia coli. AB - In a cell-free expression system derived from Escherichia coli, expression is abruptly ceased after 30 min of incubation while at this time not all the substrates have been utilized in expression. Expression-independent consumption of phosphoenolpyruvate and cysteine was found in this system, which was responsible for the above sudden cessation of expression. The above consumption was at least partially due to the dephosphorylation of nucleoside triphosphates and the conversion of cysteine into gamma-glutamylcysteine, respectively. Based on these, we developed a new system employing new S30 extract of lower phosphatase activity, higher cysteine concentration, and an inhibitor of glutathione synthesis pathway. This system showed 70% enhance in productivity (179-302 microg chloramphenicolacetyltransferase protein per ml reaction mixture per hour) over the model system. PMID- 11035185 TI - Biochemical analysis of retroviral structural proteins to identify and quantify retrovirus expressed by an NS0 murine myeloma cell line. AB - A subclone of the NS0 murine myeloma cell line, frequently used to produce recombinant monoclonal antibodies, was found by a transmission electron microscopy method to express a surprisingly high titer of 10(11) retroviral particles per ml of culture supernatant. Infectivity assays showed a very low infectious titer with the restricted host range expected for a murine amphotropic retrovirus. A Western blot assay for the viral capsid protein was developed to confirm the high titer values and provide a means for monitoring batch consistency and virus removal during the purification process. Mass spectrometry of several of the viral Gag proteins demonstrated that the cell line appeared to produce at least two closely related retroviruses. N-terminal sequencing of three of the Gag proteins demonstrated that these retroviruses were members of the murine leukemia retroviral family. Western blot detection with an antibody for the capsid protein gave a linear standard curve over the range of 0.1-3 ng per lane. This allows the detection of viral titers as low as 6x10(7) virions per ml without the need to concentrate the sample. The Western blot method has higher throughput and less variability than transmission electron microscopy methods and has potential for monitoring viral titer and clearance during development of manufacturing processes. PMID- 11035186 TI - Real time monitoring biomass concentration in Streptomyces clavuligerus cultivations with industrial media using a capacitance probe. AB - On-line monitoring biomass concentration in mycelial fed-batch cultivations of Streptomyces clavuligerus grown with soluble and partially insoluble complex media, was investigated with an in-situ capacitance probe fitted to an industrial pilot-plant tank. Standard off-line and on-line biomass determinations, including cell dry weight, packed mycelial volume, viscosity, DNA concentration and total CO(2) evolution in the exhaust gases, were performed throughout the experiments and compared to on-line capacitance measurements. Linear relations between capacitance and all other measurements were developed for both media that hold only in defined process phases, depending on the biomass state and the amount of insoluble matter present. For the industrial complex culture media good linear relations were obtained in the fast growth phase between capacitance and DNA concentration and total CO(2) evolution, while in the subsequent transition and stationary phases only with apparent viscosity was a reasonable correlation found. The capacitance probe was shown to be a valuable tool for real-time monitoring biomass concentration in industrial-like cultivation of mycelial streptomycetes. PMID- 11035187 TI - Stereospecificity of myo-inositol hexakisphosphate dephosphorylation by a phytate degrading enzyme of Escherichia coli. AB - Using a combination of high-performance ion chromatography analysis and kinetic studies, the stereospecificity of myo-inositol hexakisphosphate dephosphorylation by the phytate-degrading enzyme P2 of Escherichia coli was established. High performance ion chromatography revealed that the phytate-degrading enzyme P2 of E. coli degrades myo-inositol hexakisphosphate by stepwise dephosphorylation via D/L-Ins(1,2,3,4,5)P(5), D/L-Ins(2,3,4,5)P(4), D/L-Ins(2,4,5)P(3) or D/L Ins(1,2,4)P(3), D/L-Ins(1,2)P(2) or Ins(2, 5)P(2) or D/L-Ins(4,5)P(2) to finally Ins(2)P or Ins(5)P. Kinetic parameters for myo-inositol pentakisphosphate hydrolysis by E. coli and wheat phytase, respectively, showed that the myo inositol pentakisphosphate intermediate produced either by the phytate-degrading enzyme of wheat or E. coli are not identical. The absolute configuration of the myo-inositol pentakisphosphate isomer produced by the E. coli enzyme was determined by taking into consideration that wheat phytase produces predominantly the D-Ins(1, 2,3,5,6)P(5) isomer (Lim, P.E., Tate, M.E., 1973. The phytases: II. Properties of phytase fraction F(1) and F(2) from wheat bran and the myo-inositol phosphates produced by fraction F(2). Biochim. Biophys. Acta 302, 326-328). The data demonstrate that the phytate-degrading enzyme P2 of E. coli dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-Ins(1,2,3,4,5)P(5), D-Ins(2,3,4,5)P(4), D-Ins(2,4,5)P(3), Ins(2,5)P(2) to finally Ins(2)P (notation 6/1/3/4/5). PMID- 11035188 TI - Hydrolysis of lysergamide to lysergic acid by Rhodococcus equi A4. AB - From a mixture of lysergamide and its epimer isolysergamide, Rhodococcus equi A4 containing amidase preferentially hydrolyzed lysergamide into lysergic acid. PMID- 11035189 TI - Deletion of aprA and nprA genes for alkaline protease A and neutral protease A from bacillus thuringiensis: effect on insecticidal crystal proteins. AB - The aprA gene encoding alkaline protease A (AprA) was cloned from Bacillus thuringiensis subsp. kurstaki, and the cloned gene was used to construct aprA deleted (aprA1) strains of B. thuringiensis. An aprA1 strain of B. thuringiensis that contained the wild-type gene for neutral protease A (nprA(+)) displayed levels of extracellular proteolytic activity that were similar to those of an aprA(+)nprA(+) strain. However, when EDTA was included in the protease assay to inhibit NprA activity the aprA1nprA(+) strain displayed only 2% of the extracellular proteolytic activity of the aprA(+)nprA(+) strain. A strain that was deleted for both aprA and nprA (aprA1nprA3 strain) failed to produce detectable levels of proteolytic activity either in the presence or absence of EDTA in the assay. Compared with the aprA(+)nprA(+) strain the aprA1nprA(+) strain yielded 10% more full-length Cry1Bb crystal protein and the aprA1nprA3 strain yielded 25% more full-length Cry1Bb protein. No significant differences were seen in the 50% lethal dose of Cry1Bb protein from aprA(+)nprA(+) and aprA1nprA3 strains against three species of lepidopteran insects. These results suggest that enhanced yield of certain crystal proteins can be obtained by deletion of the genes aprA and nprA which are the major extracellular proteases of B. thuringiensis. PMID- 11035190 TI - DNA vaccination of mice with a plasmid encoding Puumala hantavirus nucleocapsid protein mimics the B-cell response induced by virus infection. AB - Inoculation of naked DNA has been applied for the development of prophylactic and therapeutic vaccines against different viral infections. To study the humoral immune response induced by DNA vaccination we cloned the entire nucleocapsid protein-encoding sequence of the Puumala hantavirus strain Vranica/Hallnas into the CMV promoter-driven expression unit of the plasmid pcDNA3, generating pcDNA3 VR1. A single dose injection of 50 microg of plasmid DNA into each M. tibialis anterior of BALB/c mice induced a high-titered antibody response against the nucleocapsid protein as documented 6 and 11 weeks after immunisation. PEPSCAN analysis of a serum pool of the pcDNA3-VR1-vaccinated animals revealed antibodies reacting with epitopes covering the whole nucleocapsid protein. The epitope specificity of the immune response induced by DNA vaccination seems to reflect the antibody response in experimentally virus-infected bank voles (the natural host of the Puumala virus) and humans. The data suggest that DNA vaccination could be used for the identification of highly immunogenic epitopes in viral proteins. PMID- 11035191 TI - Development of a p53 responsive GFP reporter; identification of live cells with p53 activity. AB - p53 is among the most intensely studied human proteins because of its vital role as the prototype tumor suppressor. As a result, there are widespread applications for p53 functional analysis in biotechnology as it relates to cancer research. p53 is a potent sequence specific transcription factor, which induces the expression of a number of genes whose products mediate cell cycle arrest and apoptosis. Because the tumor suppressor activity of p53 is dependent on its transcription regulatory function, we have undertaken to develop a p53-responsive green fluorescent protein reporter strategy to enable the identification of live cells containing p53 transcriptional transactivation activity. We demonstrate within the use of GFP fluorescence to monitor both endogenous and plasmid derived p53 biochemical and biological activity. Identifying live cells with p53 activity through GFP fluorescence will have wide application for both in vitro and in vivo studies of the p53 tumor suppressor protein. PMID- 11035192 TI - Improved resistance to transition metals of a cobalt-substituted alcohol dehydrogenase 1 from Saccharomyces cerevisiae. AB - Cobalt-substituted alcohol dehydrogenase 1 was purified from a yeast culture of Saccharomyces cerevisiae. Its reactivity towards different transition metals was tested and compared with the native zinc enzyme. The cobalt enzyme displayed a catalytic efficiency 100-fold higher than that of the zinc enzyme. Copper, nickel and cadmium exerted a mixed-type inhibition, with a scale of inhibition efficiency: Cu(2+)>Ni(2+)>Cd(2+). In general, a higher resistance of the modified protein to the inhibitory action of transition metals was observed, with two orders of magnitude for copper I(50). The presence of nickel in the complexes enzyme-coenzyme-inhibitor-substrate resulted in a decrease of the ampholytic nature of the catalytic site. On the contrary, cadmium and copper exerted an enhancement of this parameter. Electrostatic or other types of interactions may be involved in conferring a good resistance in the basic pH range, making cobalt enzyme very suitable for biotechnological processes. PMID- 11035193 TI - Preface PMID- 11035194 TI - The delivery of antisense therapeutics. AB - Antisense oligonucleotides, ribozymes and DNAzymes have emerged as novel, highly selective inhibitors or modulators of gene expression. Indeed, their use in the treatment of diseases arising from genetic abnormalities has become a real possibility over the past few years. The first antisense drug molecule is now available for clinical use in Europe and USA. However, their successful application in the clinic will require improvements in cellular targeting and intracellular delivery. This review aims to look at recent advances in the in vitro and in vivo delivery of antisense oligodeoxynucleotides and ribozymes. PMID- 11035195 TI - Selecting optimal antisense reagents. AB - Selection of the appropriate target site is crucial to the success of an antisense experiment. The selection is difficult because RNAs fold to form secondary structures, rendering most of the molecule inaccessible to intermolecular base pairing with complementary nucleic acids. Conventional approaches, such as selection by 'sequence-walking' or computer-assisted design, have not brought significant success. Several empirical selection methods have been reported, a number of which are summarised in this review. Of notable significance are the 'global' methods based on mapping of transcripts with the endoribonuclease H (RNase H) and oligonucleotide scanning arrays. PMID- 11035196 TI - Novel non-endocytic delivery of antisense oligonucleotides. AB - Antisense oligonucleotides (ONs) have several properties that make them attractive as therapeutic agents. Hybridization of antisense ONs to their complementary nucleic acid sequences by Watson-Crick base pairing is a highly selective and efficient process. Design of therapeutic antisense agents can be made more rationally as compared to most traditional drugs, i.e., they can be designed on the basis of target RNA sequences and their secondary structures. Despite these advantages, the design and use of antisense ONs as therapeutic agents are still faced with several obstacles. One major obstacle is their inefficient cellular uptake and poor accessibility to target sites. In this article, we will discuss key barriers affecting ON delivery and approaches to overcome these barriers. Current methods of ON delivery will be reviewed with an emphasis on novel non-endocytic methods of delivery. ONs are taken up by cells via an endocytic process. The process of ON release from endosomes is a very inefficient process and, hence, ONs end up being degraded in the endosomes. Thus, ONs do not reach their intended site of action in the cytoplasm or nucleus. Delivery systems ensuring a cytoplasmic delivery of ONs have the potential to increase the amount of ON reaching the target. Here, we shall examine various ON delivery methods that bypass the endosomal pathway. The advantages and disadvantages of these methods compared to other existing methods of ON delivery will be discussed. PMID- 11035197 TI - Transdermal delivery of antisense compounds. AB - Antisense technology holds tremendous promise for therapeutic applications and the study of gene function. A broadly applicable route of administration that would provide for non-invasive, simple, and convenient delivery is highly desirable. Application of oligonucleotides to the skin may represent a solution to the delivery question for both local treatment of skin disease and for systemic delivery. The iontophoretic mode of delivery for phosphorothioate oligonucleotides across hairless mouse skin reveals the potential limitation in the delivery of sufficient oligonucleotide to provide for efficacy. A potential solution to this problem is the use of significantly more potent C-5 propyne base modifications in a phosphorothioate oligonucleotide. The combination of the iontophoretic delivery mode with potent oligonucleotides resulted in selective inhibition of the CYP3A2 gene expression in the rat liver. Alternatively, oligomers with neutral charge combined with passive modes of transdermal delivery may also be feasible and represent an even more broadly applicable technology. Future studies will focus on specific applications of local and systemic therapy of antisense oligonucleotide in animal models for the design of treatment regimens. PMID- 11035198 TI - Alternative interpretations of the oligonucleotide transport literature: insights from nature. AB - Elucidation of the mechanism of oligonucleotide (ON) cellular internalization has met an impasse at the lipid penetration stage. ON internalization is commonly regarded to involve endocytosis, yet the method by which the ON penetrates the endosome membrane remains a mystery despite more than 10 years of research by multiple laboratories. In addition, the literature regarding this topic is fraught with discrepancies and inconsistencies. Therefore, the goal of this review is to propose and illustrate the feasibility of the notion that the literature discrepancies are perhaps an indication of a complex transport mechanism involving more than one uptake pathway. Accordingly, ON- and cell differences in uptake may be attributed to differences in the relative importance of these pathways for different cell types and ONs. An example of one such pathway is reviewed and critiqued in this communication with respect to its hypothetical role in ON uptake. Other innovative mechanisms should similarly be considered to stimulate new ideas, discussion and research in this unique and interesting field. PMID- 11035199 TI - Ribozyme therapy for HIV infection. AB - Within the past few years encouraging progress has been made in the treatment of HIV-1 infection, largely due to the combined use of HIV-1 protease inhibitors with nucleoside and non-nucleoside reverse transcriptase inhibitors. Despite this, HIV-1 infection is still a major global problem and the emergence of a drug resistant virus is ever present. There is a continuing need to develop new therapeutic strategies as well as improve upon all forms of existing therapies for the treatment of this viral infection. It has now been almost a decade since the first demonstration that ribozymes can effectively inhibit HIV-1 infectious spread in cell culture. Since then, ribozymes have progressed into human clinical trials primarily through gene therapy approaches. This progression brings ribozymes into the forefront as an important addition to the growing arsenal of anti-HIV-1 weapons. The following review covers the developments in anti-HIV-1 ribozyme usage over the past decade and summarizes the current state of ribozyme development for the purpose of inhibiting HIV-1 infection. PMID- 11035200 TI - Combination studies between polycationic peptides and clinically used antibiotics against Gram-positive and Gram-negative bacteria. AB - The in vitro interaction between five polycationic peptides, buforin II, cecropin P1, indolicidin, magainin II, and ranalexin, and several clinically used antimicrobial agents was evaluated against several clinical isolates of Gram positive and Gram-negative aerobic bacteria, using the microbroth dilution method. The combination studies showed synergy between ranalexin and polymyxin E, doxycycline and clarithromycin. In addition, magainin II was shown to be synergic with betalactam antibiotics. PMID- 11035201 TI - Linear and cyclic LFA-1 and ICAM-1 peptides inhibit T cell adhesion and function. AB - Short peptides derived from functional proteins have been used in several instances to inhibit activity of the parent proteins. In some cases, stability and efficacy were found to be increased by cyclization of these peptides. Inhibition of interaction of the two cell adhesion counter receptors leukocyte function-associated antigen (LFA)-1 and intercellular adhesion molecule (ICAM)-1 is being studied as a method for modulating autoimmune diseases such as rheumatoid arthritis and for facilitating organ transplantation. Here, several 10 amino acid peptides derived from the contact domains of LFA-1 and ICAM-1 were evaluated for their ability to interfere with intercellular adhesion by T cells and to inhibit a more biologic, mixed lymphocyte reaction. Both linear and cyclic forms of the peptides were effective at inhibiting intercellular adhesion. Cyclic forms were effective at inhibiting T cell activation and proliferation in the mixed lymphocyte reaction. PMID- 11035202 TI - A novel silencer element repressing expression of the GLP-1 receptor gene in fibroblasts and pancreatic A-cells, but not in pancreatic B- and D-cells. AB - The effects of the incretin hormone glucagon-like peptide 1 (7-36)amide (GLP-1) are mediated by the GLP-1 receptor (GLP-1R). This is expressed in a cell- and tissue-specific manner. Recently, we have cloned the 5'-flanking region of the human GLP-1R gene. The basal promoter activity is driven by the ubiquitous transcription factor Sp1. The tissue- and cell-specific expression of the gene requires several negatively acting cis-regulatory elements. We have now characterized one so far unknown distal cell-specific silencer element (DCS), repressing gene transcription of the human GLP-1R gene in fibroblasts and pancreatic A-cells, but not in pancreatic B- and D-cells. Our data suggests that the basal activity of the GLP-1R promoter is repressed in a tissue- and cell specific manner by this novel silencer element. PMID- 11035203 TI - Splice variants of PAC(1) receptor during early neural development of rats. AB - The specific pituitary adenylate cyclase-activating polypeptide (PACAP) receptor, PAC(1)-R, consists of at least seven isoforms, and they are differentially coupled to signal transduction pathways by alternative splicing. We have found that the major splice variants of the PAC(1) receptor seen during development are the short splice isoform, PAC(1)-R-s (which does not contain either the "hip" or "hop" cassette), and another form, PAC(1)-R-hop (which contains the "hop" cassette). We also have applied an innovative molecular histochemical technique, in situ reverse transcription-polymerase chain reaction (RT-PCR), and determined that these two splice isoforms are colocalized in the neuroepithelia from the primitive streak stage. PMID- 11035204 TI - Tissue distribution of PACAP27 and -38 in oligochaeta: PACAP27 is the predominant form in the nervous system of Lumbricus polyphemus. AB - The levels of pituitary adenylate cyclase-activating polypeptide (PACAP)27 and 38 were measured in the nervous, intestinal, excretory, and reproductive systems of Lumbricus polyphemus by radioimmunoassay. Although both PACAP27 and -38 were significantly detectable in all of the examined tissues, the distribution of the peptides was very heterogeneous. Their highest concentrations were found in the cerebral ganglia and the ventral cord, followed by the alimentary tract and the nephridial system, respectively. Moreover, the reproductive system also contained a substantial amount of PACAP. The dominant form of the peptide discovered in the majority of tissues was PACAP27. Interestingly, about 10 times more PACAP27 than PACAP38 was found, with the latter representing only a fraction of PACAP-like immunoreactivity in the tissues of Lumbricus polyphemus. PMID- 11035205 TI - Development of an antagonist of molluscan neuropeptide APGWamide with a peptide library. AB - Fifty-seven kinds of APGWamide-related peptides and a peptide library consisting of 38 peptide mixtures, each of which contained 19 kinds of APGWamide-related peptides, were synthesized with a multipeptide synthesizer, and their APGWamide agonistic or -antagonistic effects were examined on the anterior byssus retractor muscle of the bivalve Mytilus edulis and the crop of the land snail Euhadra congenita. The peptide mixtures having agonistic or antagonistic effects were subjected to HPLC purification to isolate the active peptides using the muscles as bioassay systems. Many peptides having agonistic or antagonistic effects were obtained. Of the antagonists, APGWGNamide, isolated from the peptide mixture of APGWGXamide, was the most potent. At 10(-4) M, APGWGNamide almost completely blocked the actions of 10(-6) M APGWamide on the anterior byssus retractor muscle of M. edulis and the crop of E. congenita. PMID- 11035206 TI - Fulicin regulates the female reproductive organs of the snail, Achatina fulica. AB - Fulicin is a D-amino acid-containing neuropeptide that has been thought to control male copulatory behavior in the land snail, Achatina fulica. In the present study, we demonstrated that the vagina and the oviduct of Achatina were densely innervated by fulicin-like immunoreactive neuronal fibers. We confirmed that fulicin was actually present in the vagina by mass spectrometry. Furthermore, fulicin showed a profound excitatory effect on contractions of the vagina and the oviduct. These results suggest that fulicin controls female egg laying behavior as an excitatory neuropeptide regulating the female reproductive organs of the snail. PMID- 11035207 TI - Opposing interplay between Neuropeptide FF and nitric oxide in antinociception and hypothermia. AB - This study examined the ability of the anti-opioid Neuropeptide FF (NPFF) to modify the endogenous activity of nitric oxide (NO). Antinociceptive and hypothermic effects of 1DMe (D.Tyr-Leu-(n.Me)Phe-Gln-Pro-Gln-Arg-Phe-NH(2)), an NPFF agonist, and of L-NAME (N(omega)nitro-L-arginine methyl ester), an inhibitor of nitric oxide synthase, were investigated in mice. Intraperitoneal (i.p.) injection of L-NAME induced, in the hot plate test, a dose-dependent antinociception not reversed by naloxone, an opioid antagonist, but inhibited by L-Arg, the NO synthesis precursor. Intracerebroventricular (i.c.v.) injections of 1DMe inhibit the antinociceptive activity of L-NAME in a dose-dependent manner. On the contrary, L-NAME markedly potentiated hypothermia induced by 1DMe injected in the third ventricle. These data show that Neuropeptide FF receptors exert a dual effect on endogenous NO functions and could modulate pain transmission independently of opioids. PMID- 11035208 TI - Complete inhibition of purinoceptor agonist-induced nociception by spinorphin, but not by morphine. AB - We found that spinorphin, a novel neuropeptide showed analgesia in a different manner compared with morphine. By measuring flexor responses induced by the intraplanter injection of substances, the presence of three different types of sensory neurons were demonstrated. Although spinorphin completely blocked 2 metylthioadenosine (2-MeS ATP, a P2X(3) agonist)-induced responses, morphine did not. On the other hand, morphine-induced blockade of bradykinin (BK, a B(2) receptor agonist)-responses was attenuated by pertussis toxin (PTX) treatment, whereas that of spinorphin was not. Thus it is suggested that spinorphin has a spectrum of analgesia which covers the blockade of nociception insensitive to morphine. PMID- 11035209 TI - Opioid peptide modulation of circulatory response to hyperventilation in humans. AB - After hyperventilation, systolic blood pressure (SBP) significantly decreased in 10 subjects (group 1), did not change in eight (group 2) and increased in 15 (group 3). Diastolic blood pressure and heart rate increased in all groups. The decrease in SBP was associated with a decrease in plasma catecholamines and increase in beta-endorphin, whereas the increase in SBP was accompanied by an increase in catecholamine and Met-enkephalin levels. Naloxone abolished the hyperventilation-induced SBP and catecholamine decrease only in group 1. These findings show an activation of the endogenous opioid system after hyperventilation and the role of beta-endorphin in reducing SBP in response to the test. PMID- 11035210 TI - Unlike thrombin, protein C and activated protein C do not affect vascular tone. AB - Because plasma levels of protein C (PC) or activated protein C (APC) are altered in certain diseases associated with vascular dysfunction, and APC has therapeutic potential in preventing microvascular coagulation in severe sepsis, potential vascular effects of PC and APC were compared to those of the vasoactive peptide, thrombin. Thrombin was a more potent relaxant agonist than contractile agonist in aorta. Unlike thrombin, cumulatively administered APC (10(-9)-10(-7) M) did not exert vascular effects in rat or rabbit aorta. Noncumulative challenge of PC (10( 7) M) and APC (8 x 10(-8) M) also did not contract rat or rabbit aortae, either with or without endothelium. Likewise, the same concentrations of PC and APC also did not relax norepinephrine-induced (10(-7) M) vascular tone in either rat or rabbit aortae. Thus, in contrast to thrombin, PC and APC failed to modulate vascular tone, suggesting that the therapeutic use of APC is unlikely to be accompanied by any direct effects on vascular motility. PMID- 11035211 TI - Angiotensin II does not induce apoptosis but rather prevents apoptosis in cardiomyocytes. AB - The ability of angiotensin II (ang II) to produce apoptosis is controversial. Cardiomyocytes, isolated from 7-day embryonic chick hearts and maintained in culture for 72 h, were treated with ang II. There was no evidence of ang II induced apoptosis consistently demonstrated by six different techniques: electrophoretic separation of fragmented DNA, staining with TUNEL, enzyme-linked immunosorbent assay specific for fragmented DNA, dual staining of cells with fluorescein diacetate and propidium iodide with analysis by flow cytometry, staining of nuclei with propidium iodide and cell microscopy. In contrast, apoptosis was readily induced by camptothecin or staurosporine or serum deprivation. The absence of ang II-induced cell death was also demonstrated in neonatal mouse cardiomyocytes in culture. We further sought to answer the question whether ang II Type 1 receptor blockade by antagonizing the potential beneficial effects mediated through this receptor and producing more ang II binding to the ang II Type 2 receptors, would lead to cardiac apoptosis. There was no evidence of ang II-induced apoptosis in the presence of the ang II Type 1 receptor antagonist losartan in embryonic chick cardiomyocytes. Rather ang II prevented serum deprivation-induced apoptosis. In summary, in these cardiomyocytes ang II does not induce but rather prevents apoptosis. PMID- 11035212 TI - Degradation of bradykinin by peritoneal and alveolar macrophages of the guinea pig. AB - Peptidase inhibitors and identification of the peptide fragments were used for the characterization of the bradykinin metabolism by alveolar and peritoneal macrophages. Both cell types show differences in the rate of inactivation and in the quantity of the metabolites generated. BK(1-5), BK(1-8), and BK(1-7) are the predominant direct metabolites. Metalloendopeptidase 24.15, carboxypeptidase M, and an unidentified peptidase are responsible for their formation. Angiotensin converting enzyme and neutral endopeptidase 24.11 do not play a crucial role in the degradation of bradykinin by macrophages. In the bronchoalveolar space, other cells than the macrophages are more important to the breakdown of this peptide. PMID- 11035213 TI - Potentiation of the effects of bradykinin on its receptor in the isolated guinea pig ileum. AB - Angiotensin I-converting enzyme (ACE/kininase II) inhibitors potentiated guinea pig ileum's isotonic contractions to bradykinin (BK) and its analogues, shifting the BK dose-response curve to the left. ACE inhibitors added at the peak of the contraction immediately enhanced it further (343 +/- 40%), although the ileum inactivated BK slowly (t(1/2) = 12-16 min). Chymotrypsin and cathepsin G also augmented the activity of BK up to three- or four-fold, but in a manner slower than that of ACE inhibitors. The BK B(2) receptor blocker HOE 140 inhibited all effects. Histamine and angiotensin II were not potentiated. ACE inhibitors potentiate BK independent of blocking its inactivation by inducing crosstalk between ACE and the BK B(2) receptor; proteases activate the receptor by different mechanism. PMID- 11035214 TI - Bombesin improves burn-induced intestinal injury in the rat. AB - This study was designed to determine the effect of exogenous bombesin (10 microg/kg/day, subcutaneously, three times a day) on intestinal hypomotility and neutrophil infiltration in the early and late phases of burn injury (partial thickness, second-degree burn of the skin). In acute (2 h after burn injury) or chronic (3 days after) burn groups, intestinal transit was delayed, which was reversed by bombesin treatment. In the acute burn group, but not in the chronic group, increased MPO activity was also reduced by bombesin treatment. The results demonstrate that bombesin ameliorates the intestinal inflammation due to burn injury, involving a neutrophil-dependent mechanism. PMID- 11035215 TI - The effect of alpha-melanocyte stimulating hormone on colonic inflammation in the rat. AB - The effect of alpha-melanocyte stimulating hormone (alpha-MSH) on colonic inflammation in the rat. In this study, we investigated the effects of alpha-MSH administration on trinitrobenzene sulfonic acid-induced colitis and the role of nitric oxide and prostaglandins in this response. alpha-MSH treatment (25 microg/rat, intraperitoneally; twice daily for 3 days) reduced the colonic macroscopic lesions compared to untreated ones in both acute and chronic colitis groups. This effect was reversed by pretreatment with the nitric oxide donor, sodium NP (4 mg/kg, intravenously) or cyclooxygenase-1 selective antagonist indomethacin (5 mg/kg, subcutaneously) in the acute group and with the cyclooxygenase-2 selective antagonist nimesulide (3 mg/kg, subcutaneously) in the chronic group. alpha-MSH had no effect on colonic wet weight and myeloperoxidase activity compared to the untreated colitis group. However, protein oxidation was markedly elevated in the alpha-MSH-treated group compared to untreated ones. Nitroprusside and indomethacin reversed the effect of alpha-MSH on macroscopic lesions in the acute groups, whereas nimesulide showed a similar effect in the chronic group. In conclusion, the results of our study show a protective role of alpha-MSH on colonic lesions which partially involves nitric oxide and prostaglandins. PMID- 11035216 TI - The rewarding properties of neuropeptide Y in perifornical hypothalamus vs. nucleus accumbens. AB - There is a high coexistence of substance abuse in humans with eating disorders. One theory offered to account for this fact is that a common biochemical substrate may exist that mediates both processes. Brain neuropeptide Y (NPY) is one neurochemical system that might contribute to these separate, yet related, problems. To clarify the role of NPY in mediating reward processes and the possible interaction between reward and feeding, the present study examined the effects of injecting NPY bilaterally into the perifornical hypothalamus (PFH) vs. the nucleus accumbens (NAC) on intake of preferred vs. non-preferred food types, as well as on conditioned place preference (CPP) learning. NPY (24, 78, 156 and 235 pmol/side) stimulated intake of both regular powdered chow and sucrose when injected into the PFH, but not the NAC. A CPP that was negatively correlated with food intake occurred with the low (24 pmol/side) dose of NPY in the PFH, while a CPP that was not correlated with food intake was produced with the same dose in the NAC. The extent of the CPPs produced by NPY injection in both brain sites mirrored that produced by peripheral injection of amphetamine (2.5 mg/kg). These results indicate that NPY elicits reward-related behavior, but not feeding, from the NAC, and both behaviors from the PFH. However, the feeding effect derived from the PFH appears to overshadow a rewarding effect derived from this site. Considered together, these findings suggest that altered NPY functioning in both brain regions may contribute to some of the pathophysiological processes observed in eating disordered patients who have additional proclivities for substance abuse. PMID- 11035217 TI - Secretoneurin: a functional neuropeptide in health and disease. AB - Chromogranins belong to an evolutionarily conserved family of proteins that serve as neuropeptide pro-proteins, besides having other functions. The secretogranin II-derived peptide secretoneurin is a 33-amino-acid polypeptide generated by proteolytic cleavage at paired dibasic sequences that exerts its effect by binding to specific receptors. Secretoneurin receptors have been kinetically and functionally characterized indicating that they are G-protein linked. Localization of secretoneurin and functional studies have helped to elucidate roles for secretoneurin, ranging from effects in the central nervous system to the modulation of the inflammatory response in the periphery. It has been shown that secretoneurin possesses biologic activities such as stimulation of dopamine release from striatal neurons and activation of monocyte migration, suggesting that the peptide may modulate both neurotransmission and inflammatory response. With an array of actions as diverse as that seen with other sensory neuropeptides, there is scope for numerous studies and therapeutic possibilities. PMID- 11035218 TI - Relationships among processing speed, working memory, and fluid intelligence in children. AB - The present review focuses on three issues, (a) the time course of developmental increases in cognitive abilities; (b) the impact of age on individual differences in these abilities, and (c) the mechanisms by which developmental increases in different aspects of cognition affect each other. We conclude from our review of the literature that the development of processing speed, working memory, and fluid intelligence, all follow a similar time course, suggesting that all three abilities develop in concert. Furthermore, the strength of the correlation between speed and intelligence does not appear to change with age, and most of the effect of the age-related increase in speed on intelligence appears to be mediated through the effect of speed on working memory. Finally, most of the effect of the age-related improvement in working memory on intelligence is itself attributable to the effect of the increase in speed on working memory, providing evidence of a cognitive developmental cascade. PMID- 11035219 TI - Aging and measures of processing speed. AB - Many variables have been assumed to reflect speed of processing, and most are strongly related to age in the period of adulthood. One of the primary theoretical questions with respect to aging and speed concerns the relative roles of specific and general age-related effects on particular speed variables. Distinguishing between specific (or unique) and general (or shared) age-related influences on measures of speed has been complicated, in part because the issues are sometimes framed in terms of extreme all-or-none positions, and because few researchers have employed analytical procedures suitable for estimating the relative contributions of each type of influence. However, recent methods focusing on partitioning age-related variance have indicated that large proportions of the age-related effects on individual speed variables are shared with age-related effects on other variables. Although these theoretical ideas and analytical procedures are fairly new, they may be relevant to a variety of psychophysiological or neurobiological variables. PMID- 11035220 TI - Attention and selection in the growing child: views derived from developmental psychophysiology. AB - Our understanding of developmental changes in attentional selection in the growing child has been advanced substantially by the results of (a relatively small number of) studies undertaken from a psychophysiological perspective. The basic outcome of these studies is that, in attentional filtering as well as selective set (the two basic paradigms in attention research), the processes necessary for attentional selection are in essence available even to the young child; however, the speed and efficiency of these processes tends to increase as the child grows into an adolescent. Under optimal conditions, filtering is performed at early stages of information processing, but less optimal stimulus characteristics and task requirements may induce a shift in the locus of selection to later processing stages for young children whereas older individuals are better able to preserve their early locus of selection. When early selection is constrained, young children are substantially more sensitive to the adverse effects of response competition. In selective set, sub-optimal conditions lead not so much to a shift in locus of selection processes, but to a shift in the age at which asymptote efficiency is attained. We have proposed hierarchical regression analysis as a useful technique to examine whether age-related differences in attention effects, as observed in specific ERP components and in RT, are reflections of an age effect on a single source of attentional selection or of separate sources that each contribute uniquely to the developmental trends seen in (attention effects on) RT. Re-analyses of existing data demonstrated that (again depending on task specifics) many but not all of the different component processes involved in attentional selection contributed unique variance to the age-related changes in attention effects. PMID- 11035221 TI - Age-related changes in involuntary and voluntary attention as reflected in components of the event-related potential (ERP). AB - The present paper provides an overview of age-related changes in both involuntary and voluntary attention in adult subjects as manifested in scalp-recorded ERPs. A decline in orienting with old age was inferred from a substantial reduction with age in the magnitude of deviance-related ERP components like MMN, target as well as nontarget P3s, novelty P3 and N400. A review of focused attention studies further suggested that old and young subjects do not differ substantially in the quality of attentional operations. In old subjects early selection processes, as reflected in their selection potentials, have a somewhat slower onset than in young subjects, especially in conditions in which selection is based upon complex discrimination of stimulus features. Furthermore, the global pattern emerging from visual and memory search studies is that search-related negativities in the ERPs are smaller and of longer duration in old than in young subjects over the central and anterior scalp sites. These effects could indicate that controlled search is less intense or takes more time per search operation in old than in young subjects. At more posterior scalp sites there was tendency towards an enhanced search-related negativity that could reflect a specific difficulty (or compensatory increase in mental effort) of old subjects in spatially locating targets in complex visual fields. PMID- 11035222 TI - Memory development and event-related brain potentials in children. AB - This review examines the evidence for the maturation of memory function during childhood using event-related brain potentials (ERP), and behavioral measures. It has been shown that brain structures implicated in different forms of memory mature during the first and into the second decade of life. Whereas the maturation rates of implicit and explicit memory have not been directly assessed in the literature, studies of the maturation of the corresponding brain regions imply that there should be a progression in the maturation of the different forms of memory. This review also motivates the use of brain imaging techniques for investigation of memory systems during the developing years. Although, only a handful of such studies with children are currently available, they demonstrate that such techniques can provide information that may be unavailable otherwise. For example, when children fail to generate the ERP old/new effect, an index of episodic retrieval, it has been suggested that they may lack the necessary pre existing representations in their long-term lexical or semantic memories. Similarly, age-related differences in ERP scalp topography during source memory paradigms suggest that children, who do not appear to show frontal scalp activity, lack inputs from frontal regions that are necessary for successful retrieval of source information. Future research with children will reveal more details about the nature of mnemonic processing during the developmental years. PMID- 11035223 TI - Event-related brain potential investigations of memory and aging. AB - A review of the literature that examines event-related brain potentials (ERPs) and memory with respect to aging reveals some consistency in the processes that might be spared and those that might be compromised with increasing age. By and large, the ERP repetition effect, recorded during indirect memory paradigms, appears to be relatively intact with aging, suggesting spared repetition priming mechanisms and the brain substrates upon which they depend. Some age-related findings during direct (i.e. explicit) memory testing suggest that a left-sided posterior old/new effect ( approximately 500-800 ms), thought to reflect a relatively automatic retrieval of item information, is equivalent in young and old. A later, long-duration, right-sided, prefrontal old/new effect, allied with the search for and/or the retrieval of contextual information (i.e. source memory), has been found to be smaller or absent in the waveforms of the old in two of three studies, suggesting impaired source memory mechanisms in the elderly. It is argued that the data are relatively consistent with spared item retrieval mechanisms in the elderly presumably supported by medial temporal lobe structures. However, although the data are suggestive, there are too few studies at this time to reach a firm conclusion as to whether the mechanisms that support contextual retrieval, presumably mediated by prefrontal cortical structures, are impaired in the elderly. PMID- 11035224 TI - Developmental changes in inhibitory processing: evidence from psychophysiological measures. AB - Two major theories of the development of inhibitory functioning are discussed that assume a close relation between inhibitory ability and the maturation of the frontal lobes. It is argued that a psychophysiological approach may add considerably to the study of developmental change in inhibitory processes. A selective review is presented of studies examining heart rate and brain potential measures obtained in a variety of paradigms supposedly showing inhibitory control. The results of these studies are discussed within the framework proposed by Stuss et al. [Stuss, D.T., Shallice, T., Alexander, M.P., Picton, T.W., 1995. A multidisciplinary approach to anterior attentional processing. In: Grafman, J., Holyoak, K.J., Boller, F. (Eds.), Structure and functions of the human prefrontal cortex. Ann. New York Acad. Sci. 769, 191-211], relating component processes of supervisory-system control to distinct brain regions and psychophysiological measures of attention. It is concluded that the supervisory-system framework provides a heuristic way for examining developmental changes in inhibitory processing. PMID- 11035225 TI - Structural and functional brain development and its relation to cognitive development. AB - Despite significant gains in the fields of pediatric neuroimaging and developmental neurobiology, surprisingly little is known about the developing human brain or the neural bases of cognitive development. This paper addresses MRI studies of structural and functional changes in the developing human brain and their relation to changes in cognitive processes over the first few decades of human life. Based on post-mortem and pediatric neuroimaging studies published to date, the prefrontal cortex appears to be one of the last brain regions to mature. Given the prolonged physiological development and organization of the prefrontal cortex during childhood, tasks believed to involve this region are ideal for investigating the neural bases of cognitive development. A number of normative pediatric fMRI studies examining prefrontal cortical activity in children during memory and attention tasks are reported. These studies, while largely limited to the domain of prefrontal functioning and its development, lend support for continued development of attention and memory both behaviorally and physiologically throughout childhood and adolescence. Specifically, the magnitude of activity observed in these studies was greater and more diffuse in children relative to adults. These findings are consistent with the view that increasing cognitive capacity during childhood may coincide with a gradual loss rather than formation of new synapses and presumably a strengthening of remaining synaptic connections. It is clear that innovative methods like fMRI together with MRI based morphometry and nonhuman primate studies will transform our current understanding of human brain development and its relation to behavioral development. PMID- 11035226 TI - Functional brain imaging and age-related changes in cognition. AB - There are a number of age-related structural and physiological changes in the brain that could have implications for cognitive function in the elderly. The impact of these age-related changes in the brain on cognition has been studied using neuroimaging to examine brain activity during tasks of memory, perception and attention, and determine how this activity differs between young and older individuals. It has often been found that older individuals utilize different areas of the brain than do young subjects when carrying out the same cognitive task. This has led some researchers to suggest that older persons utilize different functional brain networks, perhaps to compensate for reductions of efficiency in task-related brain areas. However, data collected to date on this issue are still limited, so although the evidence is intriguing, the definitive interpretation of these findings must await further experiments. PMID- 11035227 TI - Prevalence of Cryptosporidium infection in goats in selected locations in three agroclimatic zones of Sri Lanka. AB - The prevalence of Cryptosporidium oocysts in the faeces of 1020 goats in three age categories was examined during 1999 in selected locations of three agroclimatic zones of Sri Lanka. The oocysts were demonstrated using the Sheather's sucrose flotation method followed by staining with the modified Ziehl Neelsen technique. Cryptosporidium oocysts were detected in animals from all agroclimatic zones with the highest prevalence of infection in the dry zone (33.6%) compared with 24.7 and 21.7% in the intermediate zones and wet, respectively (P<0.001). Overall, Cryptosporidium oocyst counts were significantly higher in goats of <6 months and 7-12 months of age groups compared with goats of >12 months of age (P<0.001). Cryptosporidium oocysts were detected in 291/1020 (28.5%) animals, while 194/1020 animals (19%), 84/1020 animals (8.2%) and 13/1020 animals (1.3%) excreted low (1-1000 oocysts per gram of faeces), moderate (1000 5000 oocysts per gram of faeces) and high (>5000 oocysts per gram of faeces) counts, respectively. The mean Cryptosporidium count was 383 oocysts per gram of faeces. The majority of the infected goats were asymptomatic. These animals are likely to play an important role in the epidemiology of cryptosporidiosis in goat kids and humans. PMID- 11035228 TI - Prevalence of Cryptosporidium, Giardia and Eimeria infections in post-weaned and adult cattle on three Maryland farms. AB - The prevalence of Cryptosporidium, Giardia and Eimeria, in healthy, asymptomatic, post-weaned and mature cattle was investigated on three Maryland farms. One farm, a dairy research facility, had 150 multiparous Holstein milking cows; 24 were examined and Cryptosporidium andersoni was detected in three (12.5%) but neither Giardia nor Eimeria was detected. The second farm, a commercial dairy, had 57 multiparous Holstein milking cows and an equal number of heifers. Of 19 cows examined, C. parvum, Giardia duodenalis, and Eimeria bovis and/or E. ellipsoidalis were detected in two (10.5%), two (10.5%) and one (5.26%) cow, respectively. Of 23 heifers examined, C. parvum, Giardia, and E. bovis and E. ellipsoidalis, was detected in two (8.7%), four (17.4%), and five (21.7%), heifers, respectively. The third farm, a beef cattle breeding and genetics research facility, had 180 7- to 9-month old purebred black Angus. Of 118 examined for C. parvum and Giardia, 34 (28.8%) and 44 (37.3%) were positive, respectively, of 97 examined for E. bovis and/or E. ellipsoidalis 32 (33.0%) were positive. These findings, based on a method with a minimum detection level of 100 oocysts of C. parvum/g of feces, which underestimates the number of infected cattle, clearly demonstrate the presence of low level, asymptomatic infections in post-weaned and adult cattle in the United States and indicate the potential role of such cattle as reservoirs of infectious parasites. PMID- 11035229 TI - Culicoides arakawae (Diptera: Ceratopogonidae) population succession in relation to leucocytozoonosis prevalence on a chicken farm in Taiwan. AB - Leucocytozoonosis caused by Leucocytozoon caulleryi is a significant disease prevalent in open chicken houses of southern and eastern Asia. L. caulleryi is transmitted by Culicoides arakawae, a blood-sucking vector. Leucocytozoonosis prevalence is influenced by vector population succession. Thus this examination was performed on a farm to investigate vector population succession and leucocytozoonosis prevalence in experimental chicks and to obtain the ecology data for assessing the prevalence. The findings were as follows: (1) C. arakawae adults might be highly host specific because they were rarely discovered on cattle or pig farms, and none of the experimental chickens were infected by L. caulleryi on those farms. (2) Identifying and counting gorged and gravid C. arakawae to assess leucocytozoonosis prevalence is a practical strategy. The critical vector index should be 5.0 calculated by dividing the smallest vector mean from the prevalent period by the largest vector mean from the population not causing leucotocytozoonosis. (3) Taking vector means from three or more collections each month, should be the best assessment of leucocytozoonosis prevalence because C. arakawae succession appears to have a 3-week periodicity. Hopefully, these findings will contribute to assessing leucocytozoonosis prevalence. PMID- 11035230 TI - The specificity of antibody responses in cattle naturally exposed to Fasciola hepatica. AB - Fasciola hepatica causes significant morbidity and mortality in dairy cattle in the Andean region of Cajamarca, Peru, where prevalence of infection of up to 78% has been reported. ELISA and Western blot analyses were used to characterise antibody responses in dairy cattle to adult F. hepatica to excretory-secretory (E/S), somatic (SO) and surface (SU) antigens. Three groups of dairy cattle - calves, heifers and adult cows - naturally exposed to F. hepatica in this region, were monitored every 2 months over a 2-year period. Calves, heifers and adult cows all had antibodies which recognised a 28kDa protein in the SO preparation, whereas only adult cows had antibodies that recognised a 28kDa protein in E/S products. All three groups of cattle responded to a 60-66kDa group of proteins in E/S and SU preparations and a 17kDa antigen in SO products was recognised by antibodies from cows and heifers but not calves. The total antibody response to E/S antigens measured by ELISA, increased over time in calves and remained constantly high over the 2-year period in all three groups of cattle. Slight fluctuations in the antibody response occurred in the group of heifers and cows coinciding with seasonal changes in the level of challenge. PMID- 11035231 TI - Comparative evaluation of an indirect ELISA test for diagnosis of swine cysticercosis employing antigen from Taenia solium and Taenia crassiceps metacestodes. AB - Taenia solium cysticercosis is still a serious public health problem in several countries where poverty and lack of hygiene favor transmission. Because pigs are the primary intermediate hosts, prevalence of porcine cysticercosis is a reliable indicator of active transmission zones. Serological diagnostic methods are important tools for epidemiological studies since they can be applied to living animals on a large scale. Four antigen preparations (cyst fluid and crude) from T. solium and T. crassiceps metacestodes were compared for swine cysticercosis diagnosis by indirect ELISA (IE). Twenty-eight serum samples from swine naturally and experimentally infected by cysticerci of T. solium and 56 serum samples from swine reared in commercial herds were tested. Best results of overall sensitivity were obtained by the use of cyst fluid and crude antigen of T. crassiceps metacestode (100 and 96.4%, respectively). Using homologous antigen preparations we have observed higher specificity percentage (98.2% for cyst fluid and 96. 4% for crude metacestode T. solium antigen). We concluded that sensitivity is of far more importance than specificity for identification of endemic areas in order to prevent transmission to man. We conclude, therefore, that IE performed with cyst fluid antigen of T. crassiceps metacestode is a better tool for that purpose. PMID- 11035232 TI - The epidemiology of nematode and fluke infections in cattle in the Red River Delta in Vietnam. AB - Over a period of 13 months, faecal samples were collected monthly from approximately 45 cattle over 3 months of age. Additionally, 74 calves of 1-2 months were sampled to determine the presence of Toxocara vitulorum eggs. Individual egg counts and infective strongyle larvae from pooled faecal samples were examined. Post-mortem worm counts were carried out on six groups of tracer calves (n=12) that had been kept for 4 weeks on pasture in and around the village studied. The following helminths were identified: T. vitulorum, Cooperia punctata, C. pectinata, C. oncophora, Oesophagostomum radiatum, Trichostrongylus axei, T. colubriformis, Haemonchus spp., Fasciola spp. and Paramphistomum spp. In 8% of the samples collected from young calves, individual egg counts for T. vitulorum were found indicative for pathogenic worm burdens. Strongyle egg counts and worm counts indicated that transmission is low without a distinct seasonality. In animals of 3-9 months old, a strongyle egg count peak can be demonstrated which at a higher age steadily and significantly decreased. In faecal cultures Cooperia spp. were most prominent in all age groups throughout the year with the exception of the period September-November when Haemonchus spp. and Oesophagostomum spp. were most prevalent. Fasciola spp. eggs were found in 22% of the collected faecal samples and the egg counts were low indicating that the intensity of Fasciola spp. infection is mild. Based on the present data, regular anthelmintic treatments seem not to be justified, except for a single treatment at the age of 2 weeks against toxocariosis. PMID- 11035233 TI - Field efficacy of moxidectin in dogs and rabbits naturally infested with Sarcoptes spp., Demodex spp. and Psoroptes spp. mites. AB - The efficacy of moxidectin 1% injectable for cattle was evaluated in dogs and rabbits with naturally acquired sarcoptic, demodectic or psoroptic mites. Twenty two dogs with generalised demodicosis were orally treated with 0.4mg/kg moxidectin daily. Forty-one dogs suffering from sarcoptic mange were treated with 0.2-0.25mg/kg moxidectin either orally or subcutaneously every week for three to six times. Seven rabbits were treated orally with 0.2mg/kg moxidectin twice 10 days apart. Of the 22 dogs with demodicosis, 14% were stopped treatment because of side effects, 14% were lost and of the remaining 72% all were cured (mean therapy duration 2.4 months). Thirty-seven of the sarcoptic mange-infected dogs finished treatment and were cured. In 17% of dogs, side effects were noted. All seven rabbits treated for psoroptic mange were cured and did not show any side effect. Our results indicate that moxidectin is effective and a good alternative for the treatment of demodicosis and scabies in dogs and psoroptic mange in rabbits. Side effects seem to occur more frequently if applied subcutaneously, therefore the oral route should be preferred. PMID- 11035234 TI - CD8(+) T cell knockout mice are less susceptible to Cowdria ruminantium infection than athymic, CD4(+) T cell knockout, and normal C57BL/6 mice. AB - The role of T cells in immunity to Cowdria ruminantium was investigated by studying the responses to infection of normal, athymic, CD4(+) T cell knock out (KO) and CD8(+) T cell KO C57BL/6 mice. Normal C57BL/6 mice could be immunized by infection and treatment, and immunity was adoptively transferable from immune to naive mice by splenocytes. Following infection, athymic mice died sooner than normal mice (P=0.0017), and could not be immunized by infection and treatment. CD4(+) T cell KO mice were as susceptible to infection as normal mice and could be immunized by infection and treatment. In contrast, CD8(+) T cell KO mice were less susceptible than normal and CD4(+) T cell KO mice and 43% self-cured, while those that died did so after a prolonged incubation period. Antibody responses to C. ruminantium were CD4(+) T cell dependent, because responses were detected in immune normal and CD8(+) T cell KO mice but not in immune CD4(+) KO mice (P=0.005). Since CD8(+) T cell KO mice were less susceptible to infection, and since CD4(+) T cell KO mice could be immunized, it can be concluded that immunity to C. ruminantium can be mediated by both CD4(+) and CD8(+) T cells. PMID- 11035235 TI - Erratum to "Can an increased intake of metabolizable protein affect the periparturient relaxation in immunity against teladorsagia circumcincta in sheep?". PMID- 11035236 TI - Structural analysis of a defective transfer-like region in a staphylococcal aminoglycoside resistance plasmid. AB - Staphylococcus epidermidis clinical isolate MH6502 contained the 51. 9-kb nonconjugal plasmid pMH6502, which has homology to a major part of the transfer gene region of a known conjugal plasmid. Plasmid pMH6502 mediates aminoglycoside and ethidium bromide resistance. During restriction digest analysis of pMH6502, a double logarithmic regression of marker data gave a better linear relationship than a semi-logarithmic one. The analysis indicated several differences in the transfer-like region of pMH6502 compared to the analogous region of the S. aureus conjugal plasmid pG01. The transfer-like region was in the opposite orientation compared to pG01. An EcoRI site that is within the transfer-like region of pMH6502, has no analogue in pG01. A HindIII site, located outside a 6.3-kb EcoRI fragment in the transfer gene region of pG01, is inside the analogous fragment of pMH6502. A model is proposed to describe how a conjugal ancestral plasmid of pMH6502 could alter to its present form. PMID- 11035237 TI - Potential importance of Legionella species as etiologies in community acquired pneumonia (CAP). AB - Large percentages of patients with community acquired pneumonia (CAP) do not have a defined etiology. Between 1992-1993, 99 acute and convalescent sera were collected from patients with CAP of unknown etiology. The sera were tested using an indirect immunofluorescence antibody assay (IFA) against the following antigens: Legionella pneumophila, serogroups 3,5,6 and 7 and L. longbeachae, L. anisa, L. bozemanii and Legionella-Like Amoebal Pathogens (LLAP). A four-fold rise in titer to at least one of the antigens tested, was seen in 14% of patients; 8% to L. bozemanii, 4% to L. anisa, 2% to S. lyticum, 2% to LLAP 10 and 1% each to LLAP 1, 6 and 9. Two patients reacted to several antigens. These results indicate that other species of legionella may be important in the etiology of CAP. L. bozemanii was the organism identified in the majority of these infections. Better diagnostic studies i.e. cultures, serologies and urinary antigen testing, which recognize legionella isolates other than L. pneumophila serogroup 1 need to be developed. PMID- 11035238 TI - Comparison of Mycobacterium tuberculosis susceptibility testing performed with BACTEC 460TB (Becton Dickinson) and MB/BacT (Organon Teknika) systems. AB - The recently introduced automated culture systems MB/BacT (Organon Teknika, Belgium) was compared with radiometric BACTEC 460TB (Becton Dickinson, USA) to test antimicrobial susceptibility of Mycobacterium tuberculosis to first line drugs. On 113 strains 97.5% agreement was obtained, with the difference being not significant. Concordance was practically complete for the most important drugs, isoniazid and rifampin. The two methods however significantly differed for the time needed to complete the test; in fact MB/BacT required on the average five days more than BACTEC 460TB. Despite the delay in the completion of the test, the excellent reliability along with the elimination of radioactivity and full automation make MB/BacT an attractive alternative for susceptibility testing of M. tuberculosis. PMID- 11035239 TI - A comparison of dynamic characteristics of fluconazole, itraconazole, and amphotericin B against Cryptococcus neoformans using time-kill methodology. AB - This study evaluated the in vitro pharmacodynamics of fluconazole, itraconazole, and amphotericin B against Cryptococcus neoformans. MICs were determined for three clinical isolates according to NCCLS guidelines (M27). Time-kill studies were performed using antifungal concentrations of 0.25-32 x MIC and inocula of 10(3) and 10(5) CFU/ml. At predetermined time points over 72 hours, samples of each inoculum/drug combination were withdrawn and plated using a spiral plater. Colony counts were determined after incubation at 35 degrees C for 48 hours. Area under the kill curves (AUKCs) were plotted versus the AUC/MIC ratios. Inoculum effect was evaluated by calculating an estimated AUKC for the low inoculum then comparing it to the measured low inoculum using the unpaired Student's t-test. The MICs of fluconazole and itraconazole for isolate 97-1199, 97-1061, and 97-585 were 2, 4, 32 microg/ml and 0.03, 0.06, 0. 5 microg/ml, respectively. For amphotericin B, the MIC was 0. 25 microg/ml for each isolate. The triazoles demonstrated fungistatic activity against each isolate at both inocula with the exception of itraconazole against C. neoformans 97-585. Maximal suppression was noted at concentrations 8-16 x MIC correlating with an AUC/MIC of 192 for both inocula. Conversely, amphotericin B was fungicidal and displayed concentration dependent activity against each isolate at both inocula. Maximal killing was observed at concentrations >4 x MIC for the low inoculum and >8 x MIC for the high inoculum for each isolate. No statistically significant differences were detected between the measured and estimated AUKCs for each antifungal agent. In conclusion, our results suggest that the triazoles were most effective against C. neoformans when concentrations were maintained at 8-16 x MIC. Amphotericin B, on the other hand, was concentration-dependent; thus, greater activity was exerted at higher concentrations. PMID- 11035240 TI - Rapid identification of medically important Candida to species level by polymerase chain reaction and single-strand conformational polymorphism. AB - Invasive fungal disease has taken a great toll on immunocompromised patients. With the emergence of fluconazole and amphotericin B resistance, the rapid identification of fungi to species level is of clinical relevance in guiding appropriate antifungal therapy. Among these opportunistic fungi, Candida species are the most commonly encountered. We had developed a molecular method utilizing single-strand conformational polymorphism (SSCP) to delineate different patterns on a 260-bp amplicon from the 28S rRNA gene from six medically important Candida species. The SSCP banding patterns obtained from a total of 52 isolates were sufficiently unique to allow distinction between the species, thus indicated a high level of specificity. This method of PCR-SSCP can provide a simple and specific method for the rapid identification of medically important Candida to species level. PMID- 11035241 TI - Susceptibility testing of voriconazole, fluconazole, itraconazole and amphotericin B against yeast isolates in a Turkish University Hospital and effect of time of reading. AB - Voriconazole is a promising azole effective against a variety of fungi, including yeasts. In this study, we tested in vitro activities of voriconazole, fluconazole, itraconazole and amphotericin B against some ATCC and reference strains and 250 clinical yeast isolates. We also evaluated the effect of time of reading on MIC results. Voriconazole was the most active agent against Candida and Trichosporon isolates, including the putatively fluconazole-resistant C. krusei (MIC(90) 0.25 microg/ml) and C. glabrata (MIC(90) 0.5 microg/ml). Amphotericin B MICs were scattered in a considerably narrow range in both RPMI 1640 and Antibiotic Medium 3. MICs at 24 hours and 48 hours were similar in general for all antifungals tested. The highest percentage of strains that showed 24-hour and 48-hour MICs within +/-1-log(2) dilution was observed for amphotericin B tested in RPMI (99%), and the lowest for amphotericin B tested in Antibiotic Medium 3 (80%). In conclusion, voriconazole is very effective against a wide spectrum of Candida species and 24-hour readings could substitute 48-hour MIC evaluation. PMID- 11035242 TI - Antimicrobial activity of gemifloxacin (SB-265805), a newer fluoroquinolone, against clinical isolates of Neisseria gonorrhoeae, including fluoroquinolone resistant isolates. AB - Antimicrobial activity of gemifloxacin (SB-265805), a newly developed fluoroquinolone, to Japanese isolates of Neisseria gonorrhoeae was compared with those of various fluoroquinolones, including norfloxacin, ciprofloxacin, tosufloxacin, levofloxacin, sparfloxacin, and trovafloxacin. Among the fluoroquinolones tested, gemifloxacin was most active against N. gonorrhoeae isolates. The MIC90 values of gemifloxacin for 94 N. gonorrhoeae isolated from 1992 through 1993 and 100 isolated from 1996 through 1997 were 0.03 and 0.125 microg/ml, respectively. On the other hand, MIC90 values of the other fluoroquinolone for the 1992-1993 isolates and the 1996-1997 isolates ranged from 0.125 to 2 microg/ml and from 0.5 to 8 microg/ml, respectively. Gemifloxacin was also the most potent fluoroquinolone against 31 ciprofloxacin-resistant isolates with the ciprofloxacin MIC of 1 to 16 microg/ml, for which the gemifloxacin MIC50 and MIC90 values were 0.25 and 2 microg/ml, respectively. Moreover, the activity of gemifloxacin against fluoroquinolone-resistant gonococcal isolates containing multiple amino acid substitutions in both GyrA and ParC proteins was superior to those of the other compounds. PMID- 11035243 TI - In-vitro activity and killing effect of polycationic peptides on methicillin resistant Staphylococcus aureus and interactions with clinically used antibiotics. AB - The in-vitro activity of nisin, a 34-residue peptide produced by several Lactococcus lactis strains, and ranalexin, a 20-residue peptide isolated from the skin of the bullfrog Rana catesbeiana, alone and in combination with amoxycillin, amoxycillin-clavulanate, imipenem, clarithromycin, ciprofloxacin, rifampin and vancomycin was investigated against 40 nosocomial isolates of methicillin resistant Staphylococcus aureus (MRSA). All isolates were inhibited at concentrations of 1 to 32 microg/ml. Synergy was observed when the peptides were combined with other agents, with the exception of the beta-lactams. Finally, the consecutive exposures to each peptide did not result in selection of stable mutants with decreased susceptibility. Our finding show that nisin and ranalexin are active against MRSA, and that their activity is enhanced when they are combined with several antimicrobial agents. PMID- 11035244 TI - A novel mutation in the alpha-helix 1 of the C subunit of the F(1)/F(0) ATPase responsible for optochin resistance of a Streptococcus pneumoniae clinical isolate. AB - Previously reported mutations involved in optochin resistance of Streptococcus pneumoniae clinical isolates changed residues 48, 49 or 50, in the transmembrane alpha-helix 2 of the F(1)/F(0) ATPase subunit. We report here an unusual mutation which changes the sequence of the transmembrane alpha-helix 1 of the AtpC subunit. This mutation involves a Gly to Ser substitution resulting from a G to A transition at codon 14 of the atpC gene. PMID- 11035245 TI - Comparison of BACTEC MGIT 960 and BACTEC 460 for culture of Mycobacteria. AB - We have compared the BACTEC 460 system with the BACTEC MGIT 960 system for culture of mycobacteria from 1800 routine clinical specimens. Rate of isolation of M. tuberculosis and time to detection of positive culture was comparable for both systems (BACTEC 460, 35 isolates, BACTEC MGIT 960, 34 isolates). Contamination of cultures was more common with the BACTEC MGIT 960 system. With intensification of the decontamination process an acceptable contamination rate was achieved in the BACTEC MGIT 960 system but time to detection of positive culture was increased by 1 to 2 days. PMID- 11035246 TI - Mycobacterium intracellulare as a cause of a recurrent granulomatous tenosynovitis of the hand. AB - We report a case of recurrent granulomatous tenosynovitis with M. intracellulare in a 55-year-old HIV negative diabetic woman. Identification of the causative agent further than belonging to the M. avium-intracellulare complex is provided by specific PCR-amplification of genomic DNA and sequencing of an hypervariable region within its 16S RNA gene. Sixteen months antibiotic regimen of rifabutin and clarithromycin led to a complete resolution of the tenosynovitis. PMID- 11035247 TI - Special issue: Lars Ernster commemorative issue. PMID- 11035248 TI - In vivo control of respiration by cytochrome c oxidase in human cells. AB - The metabolic control of oxidative phosphorylation (OXPHOS) has attracted increasing attention in recent years, especially due to its importance for understanding the role of mitochondrial DNA mutations in human diseases and aging. Experiments on isolated mitochondria have indicated that a relatively small fraction of each of several components of the electron transport chain is sufficient to sustain a normal respiration rate. These experiments, however, may have not reflected the in vivo situation, due to the possible loss of essential metabolites during organelle isolation and the disruption of the normal interactions of mitochondria with the cytoskeleton, which may be important for the channeling of respiratory substrate to the organelles. To obtain direct evidence on this question, in particular, as concerns the in vivo control of respiration by cytochrome c oxidase (COX), we have developed an approach for measuring COX activity in intact cells, by means of cyanide titration, either as an isolated step or as a respiratory chain-integrated step. The method has been applied to a variety of human cell types, including wild-type and mtDNA mutation carrying cells, several tumor-derived semidifferentiated cell lines, as well as specialized cells removed from the organism. The results obtained strongly support the following conclusions: (i) the in vivo control of respiration by COX is much tighter than has been generally assumed on the basis of experiments carried out on isolated mitochondria; (ii) COX thresholds depend on the respiratory fluxes under which they are measured; and (iii) measurements of relative enzyme capacities are needed for understanding the role of mitochondrial respiratory complexes in human physiopathology. PMID- 11035249 TI - Mitochondrial energy metabolism is regulated via nuclear-coded subunits of cytochrome c oxidase. AB - A new mechanism on regulation of mitochondrial energy metabolism is proposed on the basis of reversible control of respiration by the intramitochondrial ATP/ADP ratio and slip of proton pumping (decreased H+/e- stoichiometry) in cytochrome c oxidase (COX) at high proton motive force delta p. cAMP-dependent phosphorylation of COX switches on and Ca2+-dependent dephosphorylation switches off the allosteric ATP-inhibition of COX (nucleotides bind to subunit IV). Control of respiration via phosphorylated COX by the ATP/ADP ratio keeps delta p (mainly delta psi(m)) low. Hormone induced Ca2+-dependent dephosphorylation results in loss of ATP-inhibition, increase of respiration and delta p with consequent slip in proton pumping. Slip in COX increases the free energy of reaction, resulting in increased rates of respiration, thermogenesis and ATP-synthesis. Increased delta psi(m) stimulates production of reactive oxygen species (ROS), mutations of mitochondrial DNA and accelerates aging. Slip of proton pumping without dephosphorylation and increase of delta p is found permanently in the liver-type isozyme of COX (subunit VIaL) and at high intramitochondrial ATP/ADP ratios in the heart-type isozyme (subunit VIaH). High substrate pressure (sigmoidal v/s kinetics), palmitate and 3,5-diiodothyronine (binding to subunit Va) increase also delta p, ROS production and slip but without dephosphorylation of COX. PMID- 11035250 TI - Mitochondrial free radical generation, oxidative stress, and aging. AB - Mitochondria have been described as "the powerhouses of the cell" because they link the energy-releasing activities of electron transport and proton pumping with the energy conserving process of oxidative phosphorylation, to harness the value of foods in the form of ATP. Such energetic processes are not without dangers, however, and the electron transport chain has proved to be somewhat "leaky." Such side reactions of the mitochondrial electron transport chain with molecular oxygen directly generate the superoxide anion radical (O2*-), which dismutates to form hydrogen peroxide (H2O2), which can further react to form the hydroxyl radical (HO*). In addition to these toxic electron transport chain reactions of the inner mitochondrial membrane, the mitochondrial outer membrane enzyme monoamine oxidase catalyzes the oxidative deamination of biogenic amines and is a quantitatively large source of H2O2 that contributes to an increase in the steady state concentrations of reactive species within both the mitochondrial matrix and cytosol. In this article we review the mitochondrial rates of production and steady state levels of these reactive oxygen species. Reactive oxygen species generated by mitochondria, or from other sites within or outside the cell, cause damage to mitochondrial components and initiate degradative processes. Such toxic reactions contribute significantly to the aging process and form the central dogma of "The Free Radical Theory of Aging." In this article we review current understandings of mitochondrial DNA, RNA, and protein modifications by oxidative stress and the enzymatic removal of oxidatively damaged products by nucleases and proteases. The possible contributions of mitochondrial oxidative polynucleotide and protein turnover to apoptosis and aging are explored. PMID- 11035251 TI - Persuasive evidence that quinone reductase type 1 (DT diaphorase) protects cells against the toxicity of electrophiles and reactive forms of oxygen. AB - An extensive body of evidence supports the conclusion that by catalyzing obligatory two-electron reductions of quinones to hydroquinones, NAD(P)H:quinone reductase (QR1) protects cells against the deleterious effects of redox cycling of quinones, their ability to deplete glutathione, and to produce neoplasia. The effects of elevation of QR1 levels by various enzyme inducers, inhibition of the enzyme by dicumarol, and genetic deletion of the enzyme (knockout mouse) are all consistent with the proposed protective functions. Measurement of QR1 activity in murine hepatoma cells grown in 96-well microtiter plates has provided a rapid and quantitative method for detecting inducer activity and determining inducer potency. This constitutes a strategy for the identification of potential chemoprotectors against cancer. Epidemiological studies show that humans who are genetically deficient in QR1 are more susceptible to the hematological toxicity and carcinogenicity of benzene exposure, and may be more susceptible to the development of a number of malignant tumors. PMID- 11035252 TI - Structures of mammalian cytosolic quinone reductases. AB - The metabolism of quinone compounds presents one source of oxidative stress in mammals, as many pathways proceed by mechanisms that generate reactive oxygen species as by-products. One defense against quinone toxicity is the enzyme NAD(P)H:quinone oxidoreductase type 1 (QR1), which metabolizes quinones by a two electron reduction mechanism, thus averting production of radicals. QR1 is expressed in the cytoplasm of many tissues, and is highly inducible. A closely related homologue, quinone reductase type 2 (QR2), has been identified in several mammalian species. QR2 is also capable of reducing quinones to hydroquinones, but unlike QR1, cannot use NAD(P)H. X-ray crystallographic studies of QR1 and QR2 illustrate that despite their different biochemical properties, these enzymes have very similar three-dimensional structures. In particular, conserved features of the active sites point to the close relationship between these two enzymes. PMID- 11035253 TI - Immunodetection of NAD(P)H:quinone oxidoreductase 1 (NQO1) in human tissues. AB - Despite the extensive interest in NADPH:quinone oxidoreductase (NQO1, DT diaphorase), there is little immunohistochemical information regarding its distribution in either normal human tissues or in human tumors. Using immunohistochemistry (IHC), we have examined cell-specific expression of NQO1 in many normal tissues and tumors as a step toward defining the distribution of NQO1 in humans. NQO1 was detected by IHC in respiratory, breast duct, thyroid follicle, and colonic epithelium, as well as in the corneal and lens epithelium of the eye. NQO1 was also detected by IHC in vascular endothelium in all tissues examined. NQO1 could also readily be detected in the endothelial lining of the aorta but was not detected using immunoblot analysis in the myocardium. Adipocytes stained positive for NQO1, and the enzyme was also detected by both IHC and immunoblot analysis in parasympathetic ganglia in the small intestine and in the optic nerve and nerve fibers. NQO1 was not highly expressed in five different human liver samples using immunoblot analysis, whereas studies using IHC demonstrated only trace NQO1 staining in isolated bile duct epithelium. NQO1 expresion was also examined by IHC in a variety of solid tumors. Marked NQO1 staining was detected in solid tumors from thyroid, adrenal, breast, ovarian, colon, and cornea and in non-small cell lung cancers. The NQO1 content of many solid tumors supports the use of NQO1-directed anticancer agents for therapeutic purposes, but the distribution of NQO1 in normal tissues suggests that potential adverse effects of such agents need to be carefully monitored in preclinical studies. PMID- 11035254 TI - Regulation of genes encoding NAD(P)H:quinone oxidoreductases. AB - NAD(P)H:quinone oxidoreductase (NQO1) and NRH:quinone oxidoreductase (NQO2) are flavoproteins that catalyze two-electron reduction and detoxification of quinones and its derivatives. This leads to the protection of cells against redox cycling, oxidative stress, and neoplasia. NQO1 is expressed ubiquitously in all the tissues. However, the level of expression varied among the human tissues. NQO1 gene is expressed at higher levels in several tumor tissue types, including liver and colon, as compared to normal tissues of similar origin. NQO1 gene expression is coordinately induced with other detoxifying enzyme genes in response to xenobiotics, antioxidants, oxidants, heavy metals, and radiations. Deletion mutagenesis in the NQO1 gene promoter identified several cis-elements including antioxidant response element (ARE), a basal element, and AP-2 element. ARE elements have also been found in the promoter regions of other detoxifying enzyme genes including glutathione S-transferases. ARE is essentially required for expression and coordinated induction of NQO1 and other detoxifying enzyme genes. Nuclear transcription factors Nrf2 and c-Jun bind to the ARE and activate the gene expression. The binding of Nrf2 + c-Jun to the ARE required unknown cytosolic factor(s). In addition to Nrf2 and c-Jun, other nuclear transcription factors including Nrf1, Jun-B, and Jun-D also bind to the ARE and regulate expression and induction of NQO1 gene. A hypothetical model is presented based on the available information on ARE-mediated regulation of detoxifying enzyme genes. Briefly, the Nrf2 is retained in the cytosplasm by a repressor protein Keap1 in untreated normal cells. The treatment of cells with xenobiotics and antioxidants leads to the activation of unknown cytosolic factor(s) that catalyze modification of Nrf2 and/or Keap1. The modification follows dissociation of Nrf2 and Keap1. The free Nrf2 translocates in the nucleus. Nrf2 in the nucleus heterodimerizes with c-Jun and binds to the ARE resulting in the induction of NQO1 and other ARE regulated genes expression. The identity of cytosolic factor(s) remains unknown. PMID- 11035255 TI - The role of NAD(P)H oxidoreductase (DT-Diaphorase) in the bioactivation of quinone-containing antitumor agents: a review. AB - Bioactivation of quinone-containing anticancer agents has been studied extensively within the context of the chemistry and structure of the individual quinones which may result in various mechanisms of bioactivation and activity. In this review we focus on the two electron enzymatic reduction/activation of quinone-containing anticancer agents by DT Diaphorase (DTD). This enzyme has become important in oncopharmacology because its activity varies with tissues and it has been found to be elevated in tumors. Thus, a selective tumor cell kill can exist for agents that are good substrates for this enzyme. In addition, the enzyme can be induced by a variety of agents, a fact that can be used in chemotherapy. That is induction by a nontoxic agent followed by treatment with a good DT-Diaphorase substrate. A wide variety of anticancer drugs are discussed some of which are not good substrates such as Adriamycin, and some of which are excellent substrates. The latter category includes a variety of quinone containing alkylating agents. PMID- 11035256 TI - Structure-function studies of DT-diaphorase (NQO1) and NRH: quinone oxidoreductase (NQO2). AB - DT-diaphorase, also referred to as NQO1 or NAD(P)H: quinone acceptor oxidoreductase, is a flavoprotein that catalyzes the two-electron reduction of quinones and quinonoid compounds to hydroquinones, using either NADH or NADPH as the electron donor. NRH (dihydronicotinamide riboside): quinone oxidoreductase, also referred to as NQO2, has a high nucleotide sequence identity to DT diaphorase and is considered to be an isozyme of DT-diaphorase. These enzymes transfer two electrons to a quinone, resulting in the formation of a hydroquinone product without the accumulation of a dissociated semiquinone. Steady and rapid reaction kinetic experiments have been performed to determine the reaction mechanism of DT-diaphorase. Furthermore, chimeric and site-directed mutagenesis experiments have been performed to determine the molecular basis of the catalytic differences between the two isozymes and to identify the critical amino acid residues that interact with various inhibitors of the enzymes. In addition, functional studies of a natural occurring mutant Pro-187 to Ser (P187S) have been carried out. Results obtained from these investigations are summarized and discussed. PMID- 11035257 TI - Regulation of ubiquinone metabolism. AB - Interest in ubiquinone (UQ) has increased during recent years, mainly because of its antioxidant function and its use as a dietary supplement. However, our knowledge of the biosynthesis, catabolism, and regulation of this lipid in mammalian tissues is quite limited. UQ exhibits a high rate of turnover in all tissues indicating that cells possess efficient metabolic pathways for handling this compound and controlling its tissue levels. Besides reviewing the generally accepted metabolic pathway, alternative synthetic mechanisms are described. The lack of data concerning catabolism and regulation of this compound is emphasized. Reasons for the rather limited uptake of dietary UQ are discussed and alternative mechanisms for its beneficial effects on organ function are suggested. Since appropriate tissue uptake of dietary UQ probably only occurs in deficient states, the definition of partial UQ deficiency and its consequences is urgently needed. The possibility of raising tissue UQ levels by drug treatment or natural metabolites is raised as a choice of preference for the future. PMID- 11035258 TI - Anti-atherogenic effect of coenzyme Q10 in apolipoprotein E gene knockout mice. AB - Oxidation of low-density lipoprotein (LDL) lipid is implicated in atherogenesis and certain antioxidants inhibit atherosclerosis. Ubiquinol-10 (CoQ10H2) inhibits LDL lipid peroxidation in vitro although it is not known whether such activity occurs in vivo, and, if so, whether this is anti-atherogenic. We therefore tested the effect of ubiquinone-10 (CoQ10) supplemented at 1% (w/w) on aortic lipoprotein lipid peroxidation and atherosclerosis in apolipoprotein E-deficient (apoE-/-) mice fed a high-fat diet. Hydroperoxides of cholesteryl esters and triacylglycerols (together referred to as LOOH) and their corresponding alcohols were used as the marker for lipoprotein lipid oxidation. Atherosclerosis was assessed by morphometry at the aortic root, proximal and distal arch, and the descending thoracic and abdominal aorta. Compared to controls, CoQ10-treatment increased plasma coenzyme Q, ascorbate, and the CoQ10H2:CoQ10 + CoQ10H2 ratio, decreased plasma alpha-tocopherol (alpha-TOH), and had no effect on cholesterol and cholesterylester alcohols (CE-OH). Plasma from CoQ10-supplemented mice was more resistant to ex vivo lipid peroxidation. CoQ10 treatment increased aortic coenzyme Q and alpha-TOH and decreased the absolute concentration of LOOH, whereas tissue cholesterol, cholesteryl esters, CE-OH, and LOOH expressed per bisallylic hydrogen-containing lipids were not significantly different. CoQ10 treatment significantly decreased lesion size in the aortic root and the ascending and the descending aorta. Together these data show that CoQ10 decreases the absolute concentration of aortic LOOH and atherosclerosis in apoE-/- mice. PMID- 11035259 TI - Lipid peroxidation in membranes and low-density lipoproteins: similarities and differences. AB - Lipid peroxidation has been a central aspect of studies of the nature of free radical species and their origin in biological systems. Moreover, there has been a growing interest in lipid peroxidation based on evidence that biologically active products are formed that influence cell function and the course of major human diseases. A review of the work in this area is contributed by Lars Ernster is presented with an emphasis on the mechanisms by which lipid peroxidation is initiated in biological lipid systems. Based on what was described for metal catalyzed oxidation of cell membranes, and the seminal studies on cytochrome P 450-mediated lipid peroxidation, several parallel and distinct aspects of lipid peroxidation are described. A key distinction between lipid peroxidation in cell membranes and lipoproteins reveals aspects of free radical initiated reactions involving proteins and lipids that determine pro- vs. anti-oxidant outcomes, and the role of lipid structure and order in delineating the progress of oxidation. PMID- 11035260 TI - The role of the redox protein thioredoxin in cell growth and cancer. AB - The thioredoxins are ubiquitous proteins containing a conserved -Trp-Cys-Gly-Pro Cys-Lys- redox catalytic site. Mammalian thioredoxin family members include thioredoxin-1 (Trx1), mitochondrial thioredoxin-2 (Trx2), and a larger thioredoxin-like protein, p32TrxL. Thioredoxin is reduced by NADPH and thioredoxin reductase and, in turn reduces oxidized cysteine groups on proteins. When thioredoxin levels are elevated there is increased cell growth and resistance to the normal mechanism of programmed cell death. An increase in thioredoxin levels seen in many human primary cancers compared to normal tissue appears to contribute to increased cancer cell growth and resistance to chemotherapy. Mechanisms by which thioredoxin increases cell growth include an increased supply of reducing equivalents for DNA synthesis, activation of transcription factors that regulate cell growth, and an increase in the sensitivity of cells to other cytokines and growth factors. The mechanisms for the inhibition of apoptosis by thioredoxin are just now being elucidated. Because of its role in stimulating cancer cell growth and as an inhibitor of apoptosis, thioredoxin offers a target for the development of drugs to treat and prevent cancer. PMID- 11035261 TI - Triggering and modulation of apoptosis by oxidative stress. AB - Cell survival requires multiple factors, including appropriate proportions of molecular oxygen and various antioxidants. Although most oxidative insults can be overcome by the cell's natural defenses, sustained perturbation of this balance may result in either apoptotic or necrotic cell death. Numerous, recent studies have shown that the mode of cell death that occurs depends on the severity of the insult. Oxidants and antioxidants can not only determine cell fate, but can also modulate the mode of cell death. Effects of oxidative stress on components of the apoptotic machinery may mediate this modulation. This review will address some of the current paradigms for oxidative stress and apoptosis, and discuss the potential mechanisms by which oxidants can modulate the apoptotic pathway. PMID- 11035262 TI - Separation of cytochrome c-dependent caspase activation from thiol-disulfide redox change in cells lacking mitochondrial DNA. AB - Release of mitochondrial cytochrome c (cyt c) is an early and common event during apoptosis. Previous studies showed that the loss of cyt c triggered superoxide production by mitochondria and contributed to the oxidation of cellular thiol disulfide redox state. In this study, we tested whether loss of the functional electron transport chain due to depleting mitochondrial DNA (mtDNA) would affect this redox-signaling mechanism during apoptosis. Results showed that cyt c release and caspase activation in response to staurosporine treatment were preserved in cells lacking mitochondrial DNA (rho0 cells). However, unlike the case with rho+ cells, in which a dramatic oxidation of intracellular glutathione (GSH) occurred after mitochondrial cyt c release, the thiol-disulfide redox state in apoptotic rho0 cells remained largely unchanged. Thus, mitochondrial signaling of caspase activation can be separated from the bioenergetic function, and mitochondrial respiratory chain is the principal source of ROS generation in staurosporine-induced apoptosis. PMID- 11035263 TI - Peroxynitrite formed by mitochondrial NO synthase promotes mitochondrial Ca2+ release. AB - Mitochondria contribute to the maintenance of the intracellular Ca2+ homeostasis by taking up and releasing the cation via separate and specific pathways. The molecular details of the release pathway are elusive but its stimulation by the cross-linking of some vicinal thiols and consequently NAD+ hydrolysis are known. Thiol cross-linking and NAD+ hydrolysis can be achieved by addition of peroxynitrite (ONOO-), the product of the reaction between superoxide (O2-) and nitric oxide (nitrogen monoxide, NO*) to mitochondria. Mitochondria contain an NO synthase (mtNOS), which is stimulated by Ca2+, and are a copious source of O2-. We show here that intramitochondrially formed ONOO- stimulates the specific, NAD+ linked Ca2+ release from mitochondria. Our findings that upon Ca2+ uptake mtNOS is stimulated, that ONOO- is formed, and that Ca2+ is subsequently released from intact mitochondria suggest the existence of a feedback loop, which prevents overloading of mitochondria with Ca2+. PMID- 11035264 TI - Reactions of peroxynitrite in the mitochondrial matrix. AB - Superoxide radical (O2-) and nitric oxide (NO) produced at the mitochondrial inner membrane react to form peroxynitrite (ONOO-) in the mitochondrial matrix. Intramitochondrial ONOO- effectively reacts with a few biomolecules according to reaction constants and intramitochondrial concentrations. The second-order reaction constants (in M(-1) s(-1)) of ONOO- with NADH (233 +/- 27), ubiquinol-0 (485 +/- 54) and GSH (183 +/- 12) were determined fluorometrically by a simple competition assay of product formation. The oxidation of the components of the mitochondrial matrix by ONOO- was also followed in the presence of CO2, to assess the reactivity of the nitrosoperoxocarboxylate adduct (ONOOCO2-) towards the same reductants. The ratio of product formation was about similar both in the presence of 2.5 mM CO2 and in air-equilibrated conditions. Liver submitochondrial particles supplemented with 0.25-2 microM ONOO- showed a O2- production that indicated ubisemiquinone formation and autooxidation. The nitration of mitochondrial proteins produced after addition of 200 microM ONOO- was observed by Western blot analysis. Protein nitration was prevented by the addition of 50 200 microM ubiquinol-0 or GSH. An intramitochondrial steady state concentration of about 2 nM ONOO- was calculated, taking into account the rate constants and concentrations of ONOO- coreactants. PMID- 11035265 TI - A simpler, more robust method for the analysis of 8-oxoguanine in DNA. AB - The oxidized DNA base 8-oxoguanine has been commonly measured by enzymatic digestion of DNA to nucleosides followed by high-performance liquid chromatography (HPLC) separation of the adduct 8-oxodeoxyguanosine. There has recently been an enormous debate surrounding the validity of this approach, from which it has become clear that artifactual oxidation of the native base to 8 oxoguanine can occur at numerous stages in sample preparation. Hence, we have designed an alternative protocol to traditional enzymatic digestion of DNA which (i) limits the potential for artifactual oxidation, (ii) speeds up the assay markedly, (iii) increases the assay's sensitivity moderately, and (iv) addresses criticisms that have been raised concerning the efficiency of DNA digestion by nucleases. In short, we use the Escherichia coli repair enzyme formamidopyrimidine (Fapy) glycosylase to release the base 8-oxoguanine from full length DNA, then separate 8-oxoguanine from high molecular weight molecules by ultrafiltration (10,000 Da exclusion) and analyze the base adduct by reverse phase HPLC. Benefits of this approach include (i) rapid removal of the roughly million-fold molar excess of unaltered bases from the sample, (ii) reduction in the length of enzymatic incubations and the number of steps, (iii) elimination of high temperature incubation, (iv) a very clean chromatographic separation, and (v) rapid elution of the analyte and correspondingly greater throughput. Using this improved method, we have followed the induction of 8-oxoguanine in the DNA of peroxide-treated HeLa cells, an experiment that had proved cumbersome with traditional methods. PMID- 11035266 TI - Ascorbate-dependent recycling of the vitamin E homologue Trolox by dihydrolipoate and glutathione in murine skin homogenates. AB - In the redox antioxidant network, dihydrolipoate can synergistically enhance the ascorbate-dependent recycling of vitamin E. Since the major endogenous thiol antioxidant in biological systems is glutathione (GSH) it was of interest to compare the effects of dihydrolipoate with GSH on ascorbate-dependent recycling of the water-soluble homologue of vitamin E, Trolox, by electron spin resonance (ESR). Trolox phenoxyl radicals were generated by a horseradish peroxidase (HRP) hydrogen peroxide (H2O2) oxidation system. In the presence of dihydrolipoate, Trolox radicals were suppressed until both dihydrolipoate and endogenous levels of ascorbate in skin homogenates were consumed. Similar experiments made in the presence of GSH revealed that Trolox radicals reappeared immediately after ascorbate was depleted and that GSH was not able to drive the ascorbate-dependent Trolox recycling reaction. However, at higher concentrations GSH was able to increase ascorbate-mediated Trolox regeneration from the Trolox radical. ESR and spectrophotometric measurements demonstrated the ability of dihydrolipoate or GSH to react with dehydroascorbate, the two-electron oxidation product of ascorbate in this system. Dihydrolipoate regenerated greater amounts of ascorbate at a much faster rate than equivalent concentrations of GSH. Thus the marked difference between the rate and efficiency of ascorbate generation by dihydrolipoate as compared with GSH appears to account for the different kinetics by which these thiol antioxidants influence ascorbate-dependent Trolox recycling. PMID- 11035267 TI - Potential health impacts of excessive flavonoid intake. AB - Plant flavonoids are common dietary components that have many potent biological properties. Early studies of these compounds investigated their mutagenic and genotoxic activity in a number of in vitro assays. Recently, a renewed interest in flavonoids has been fueled by the antioxidant and estrogenic effects ascribed to them. This has led to their proposed use as anticarcinogens and cardioprotective agents, prompting a dramatic increase in their consumption as dietary supplements. Unfortunately, the potentially toxic effects of excessive flavonoid intake are largely ignored. At higher doses, flavonoids may act as mutagens, pro-oxidants that generate free radicals, and as inhibitors of key enzymes involved in hormone metabolism. Thus, in high doses, the adverse effects of flavonoids may outweigh their beneficial ones, and caution should be exercised in ingesting them at levels above that which would be obtained from a typical vegetarian diet. The unborn fetus may be especially at risk, since flavonoids readily cross the placenta. More research on the toxicological properties of flavonoids is warranted given their increasing levels of consumption. PMID- 11035269 TI - Early total white blood cell recovery is a predictor of low number of apheresis and good CD34(+) cell yield. AB - OBJECTIVE: We analysed peripheral blood progenitor cell (PBPC) mobilisation and collection in order to assess the main factors related to CD34(+) cell yields in patients affected by haematological malignancies. PATIENTS AND METHODS: The features of CD34(+) cell mobilisation of patients with haematological malignancies that underwent autologous bone marrow transplantation were examined. Mobilisation chemotherapy consisted mainly of cyclophosphamide (CY) 4 or 7 g/m(2) followed by growth factors. Leukapheresis was started when the WBC counts reached 1.0x10(9)/l with the aim to collect at least 5x10(6) CD34(+) cells/kg body weight. The aphereses were performed on continuous-flow blood cell separators. The analysed variables were: age, diagnosis, CT mobilisation regimen, type of growth factor, number of previous CT lines, prior radiotherapy, days for WBC recovery and number of aphereses procedures to achieve the target of CD34(+) cells. RESULTS: There were 41 consecutive patients (26 M/15 F): 21 non-Hodgkin's lymphoma (NHL), 15 Hodgkin's disease (HD), two chronic myeloid leukaemia (CML) and three multiple myeloma (MM). Eleven patients could not collect the proposed threshold of CD34(+) cells. CY 4 mobilised patients recovered WBC counts in less days (P=0.03). By ANOVA, the days to WBC recovery had a linear function of the predictors "number of aphereses" and "type of mobilisation CT" (coefficients: 0.86 and 0.95, respectively). For the number of aphereses and WBC recovery after CT mobilisation, we obtained a correlation coefficient of 0.36 (P=0.02). CONCLUSION: This study shows that it is feasible to mobilise and collect PBPC in patients previously treated with CT with or without RT. There was a linear correlation between the days for WBC recovery and the number of aphereses needed to collect the target number of CD34(+) cells. The study suggests that early WBC recovery, using mainly CY 4 mobilisation chemotherapy, is an important predictor of a low number of aphereses to achieve a good CD34(+) yield. PMID- 11035270 TI - Difficulties in the management of an incomplete form of refractory thrombotic thrombocytopenic purpura, the usefulness of vincristine. AB - BACKGROUND: Although several etiologies can be identified in thrombotic thrombocytopenic purpura (TTP), idiopathic cases are still frequent. Incomplete forms are more rare. Currently, the diagnosis may be made in cases of thrombocytopenia and microangiopathic hemolytic anemia. According to the literature, mortality and morbidity are significantly improved with plasma exchange. However, treatment in refractory forms remains problematic. CASE REPORT: A 33-year old woman presented with an incomplete form of TTP, refractory to a combination of therapeutics. The patient underwent plasma infusion, plasma exchange, and then was started on corticosteroids. She also received intravenous immunoglobulins and antiplatelet agents in close proximity to vincristine (Oncovin) infusion. The main biological indicators used were the platelet count, hematocrit, LDH, and the presence of schistocytes. Following vincristine treatment, the patient's condition rapidly improved. CONCLUSION: Vincristine administered after the failure of standard therapeutics was effective in this refractory form of TTP. PMID- 11035272 TI - Leucodepletion process performance variation: is it dependent on the batch of filters, the processing centre or the counting technology? AB - The aim of this retrospective study was to determine whether variations in the enumerated white cell contamination of leucocyte-depleted products was caused by the filter batch, the processing centre or by counting technology related issues. The influence of donor variation is also considered. The results suggest that for some red cell processes, variation is mainly the result of counting technology differences. Other products do not display similar trends though all leucodepletion processes may give rare high white count failures due to donor related issues, though defective filter batch cannot be excluded requiring continual review. PMID- 11035271 TI - Three episodes of delayed hemolytic transfusion reactions due to multiple red cell antibodies, anti-Di, anti-Jk and anti-E. AB - There is no report in which three episodes of delayed hemolytic transfusion reaction (DHTR) occurred from multiple antibodies to red cells (RBCs) in the course of treatment of a patient. This paper describes episodes of anemia and hyperbilirubinemia in concert with the development of three alloantibodies in a multiple transfused patient. The patient was a 71-year-old male suffering from valvular heart disease and hemophilia B with a history of transfusions. Although he received compatible RBCs from 14 donors as judged by a crossmatch test using the albumin-antiglobulin method, three episodes of DHTR occurred after surgery. The first hemolytic episode on day 7 after surgery was due to anti-Di(a) because of clinical and laboratory evidence which included jaundice, sudden increases in total bilirubin (T-Bil) and lactate dehydrogenase (LD) levels, and a decrease (2.2 g/dl) in hemoglobin (Hb) level. The second hemolytic episode on day 16 resulted from newly producted anti-Jk(b). The patient experienced fever, fatigue, nausea and anorexia, and laboratory data showed a second increase in T-Bil, a second decrease (3 g/dl) in Hb, and moderate elevations of blood urea nitrogen (BUN) and creatinine (CRE) levels. The third hemolytic episode on day 39 was due to anti-E. The patient complained of fever and fatigue and had a third unexplained drop (1.5 g/dl) in Hb despite no bleeding. This is the first reported case in which three episodes of DHTR occurred from different red cell antibodies. PMID- 11035273 TI - Haematopoietic stem cell transplantation. PMID- 11035274 TI - Peripheral blood stem cell collection: the interaction of technology, procedure, and biological factors. AB - Centrifugal technology, continuous flow and discontinuous flow, has served as the technology platform for extracting cell concentrates of interest from peripheral blood (PB) for patient therapy for the past 35-40 yr. Models for procedure outcome exist for collection of normal donor (ND) platelet and granulocyte concentrates that integrate: (1) biological variables (pre-procedure PB cell concentration, the total circulating quantity of cells, donor/patient blood volume (BV)), (2) device efficiency, and (3) procedure parameters such as total blood processed (TBP), and in the case of cytoreductions - the volume collected. (cf. Hester J, Kellogg R, Mulzet A, et al., Blood (54) (1979) 254; Hester J, Ventura G, J Clin Apheresis (4) (1988) 188.) To date, no predictive CD34+ yield algorithm integrating these three variables has been formulated that could be applied prospectively for individual ND or patients (PT). There are economic, toxicity and statistical comparison benefits to be derived from generating such an algorithm.A small pilot study is presented with a brief review of current publications that suggest the circulating quantity of CD34+ cells available to be collected and the quantity mobilized during leukapheresis are the major contributing factors to CD34+ yield, somewhat obscuring the role of the total blood processed (TBP). Intraprocedure CD34+ cell mobilization, incompletely characterized to date, appears to be a dynamic nonlinear process, as the harvested yield does not rise proportionally as the fraction of BV processed increases. And, like the pre-procedure PB CD34+ concentration and total circulating quantity, CD34+ mobilization during leukapheresis probably relates to prior treatment and the priming regimen. Studies that provide: (1) separate analyses of PT populations divided according to chemotherapy toxicity factors; (2) design and implementation of optimal priming regimens with respect to dose 'intensity' of both growth factors and chemotherapy; and (3) standardization of laboratory assays of CD34+ enumeration seem essential to generating a predictive algorithm. PMID- 11035275 TI - Adoptive allogeneic immunotherapy--history and future perspectives. AB - For more than 30 yrs allogeneic hematopoietic stem cell transplantations have been successfully performed in patients with hematologic malignancies and bone marrow aplasia. Over the years the field of transplantation has changed dramatically. More and more unrelated donors became available, regimens for haploidentical transplantations were introduced and G-CSF mobilized peripheral blood stem cells and fetal cells from umbilical cord became available as alternate sources of hematopoietic stem cells. However, especially the introduction of donor lymphocyte infusions (DLI) for the successful treatment of leukemic relapses after allogeneic stem cell transplantations improved our understanding of transplantation immunology and opened amazing perspectives in allogeneic transplantation. It was long believed, that myeloablative therapy with high-dose chemotherapy and total body irradiation (TBI) are the sole antileukemic principles in allogeneic transplantations. But by now it became clear, that donor lymphocytes exert a very potent antileukemic effect, now referred as the graft versus-leukemia (GVL) or graft-versus-malignancy (GVM) reaction. The efficacy of DLI in controlling leukemic relapses suggests that myeloablative therapy is not essential for long-term disease control. By exploiting the GVL or GVM reaction more intensively the role of chemotherapy and TBI is changing to immunosuppression. Sufficient immunosuppression to allow grafting, however, can be achieved with much lower doses as those which have been used in conventional transplants. Therefore allogeneic transplants have become also available for the elderly or for patients with concurrent medical conditions, which would have excluded them from conventional transplants. Moreover, this allogeneic transplantation strategy with reduced intensity conditioning is now also under investigation in patients with susceptible solid tumors and autoimmune diseases. However, one major obstacle in allogeneic transplantations, namely the graft versus-host disease (GVHD), remains to be solved. PMID- 11035276 TI - T cell depletion of allogeneic stem cell grafts with anti-CD 52 monoclonal antibodies: the Ulm experience from 1983-1999. PMID- 11035277 TI - IVIG as a nonspecific accelerator of restitutio ad integrum. PMID- 11035278 TI - Antenatal steroid treatment and adverse fetal effects: what is the evidence? AB - This article reviews current animal and human data regarding possible adverse fetal effects from antenatal steroid treatment. Although it is now well accepted that such treatment is of benefit to fetal lung development, the potential for adverse fetal outcomes as a result of single or multiple glucocorticoid dosing has not been widely recognized. There are now growing concerns, based on animal and some human data, that repeated antenatal doses could lead to a decrease in birth weight, a decrease in fetal brain and other organ size, and abnormal neuronal development. Previous investigations have been hampered by nonstandardization in the type of glucocorticoid, route of delivery, timing of administration, and number of treatment courses. It is recommended that these concerns be addressed through large randomized, controlled clinical trials. In the meantime, it would be prudent to minimize antenatal steroid treatments to a single course with repeated dosing only if there is a persistent threat of preterm delivery. The practice of giving weekly injections of steroids starting at fetal viability and continuing into the third trimester is not supported. PMID- 11035279 TI - Chronic hypoxia and developmental regulation of cytochrome c expression in rats. AB - OBJECTIVE: To test the hypothesis that chronic hypoxia upregulates cytochrome c expression in heart, brain, and liver of fetal and maternal rats. METHODS: Time dated pregnant Sprague-Dawley rats were divided into normoxic and hypoxic (48 hours of 10.5% oxygen from days 19 to 21) groups, and were killed on day 21. Tissue levels of cytochrome c in heart, brain, and liver were determined by using monoclonal antiserum for cytochrome c. RESULTS: Chronic hypoxia caused a decrease in fetal body weight (5.3 +/- 0.1 to 4.7 +/- 0.1 g) and an increase in heart/body weight ratio (0.0048 +/- 0.0001 to 0.0061 +/- 0.0002). Cytochrome c levels were 4 , 2.6-, and 13-fold higher in heart, liver, and brain, respectively, of the mother than of the fetus. Chronic hypoxia did not change cytochrome c levels in maternal tissues but caused a 70% increase and 54% decrease in cytochrome c levels in the fetal heart and liver, respectively. No difference was observed in the fetal brain. CONCLUSIONS: The results suggest that expression of cytochrome c is tissue specific and developmentally regulated. Chronic hypoxia showed differential regulation of cytochrome c levels both developmentally and tissue specifically. The increased sensitivity of cytochrome c in fetal tissue to chronic hypoxia is likely to represent a fetal adaptive mechanism to the stress of chronic hypoxia. PMID- 11035280 TI - Maturation of ovine uterine smooth muscle during development and the effects of parity. AB - OBJECTIVES: To characterize changes in myometrial contractile proteins and myosin heavy chain (MHC) isoforms during ovine fetal and neonatal development and after pregnancy. We hypothesized that ovine myometrium demonstrates progressive cellular differentiation and maturation which begins in utero and extends into the postnatal period, and that pregnancy causes further cellular alterations. METHODS: Myometrium was obtained from female fetal (72- to 140-days of gestation, n = 19; term = approximately 145 days), postnatal (1 day to 3 months, n = 25), and parous noncycling nonpregnant (n = 9) sheep to measure total and soluble proteins, actin, MHC, and MHC isoforms. Contractile proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and expression of 200 kD MHC isoforms were determined with Western immunoblots. RESULTS: The contents of total and soluble proteins and actin and total myosin gradually increase (P <.003) during ovine development. Although the contribution of smooth-muscle 204 kD MHC increased (P <.001) from 23 +/- 8% of total MHC at <100 days of gestation to 75 +/- 2% 3 to 4 months postnatally, the 200-kD species fell proportionately. Before birth, MHC-B, a fetal isoform, is the predominant 200-kD protein; postnatally, it is replaced by SM2, demonstrating a switch from a synthetic to a mature contractile smooth-muscle phenotype. Pregnancy is associated with further increases in actin contents and redistribution of the contents of the 204-kD and SM2 MHC isoforms. CONCLUSIONS: Although the fetal and postnatal uterus has no known functional demand, ovine myometrial differentiation and maturation begin in the midtrimester and continue throughout the postnatal period. Thus, changes in smooth-muscle phenotype occur prenatally, as evidenced by a switch from MHC-B to SM2, which may signal completion of organ development and preparation for adult function. Pregnancy results in further modifications in myometrial proteins. PMID- 11035281 TI - Regulation of activin A, inhibin A, and follistatin production in human amnion and choriodecidual explants by inflammatory mediators. AB - OBJECTIVE: To determine the effects of inflammatory mediators on the production of activin A, inhibin A, and the binding protein follistatin in term amnion and choriodecidual tissues. METHODS: The effects of interleukin-1 beta (IL-1 beta; 1 ng/mL), tumor necrosis factor-alpha (TNF-alpha; 10 ng/mL), and bacterial lipopolysaccharide (LPS; 5 microg/mL) on production rates of activin A, inhibin A, and follistatin by term choriodecidual and amnion membranes in explant culture were determined using specific enzyme-linked immunoabsorbent assays. RESULTS: All explants (n = 6 placentas) produced detectable amounts of activin A, inhibin A, and follistatin under basal conditions; choriodecidual production rates were more than tenfold higher than amnion rates. In amnion explants, activin A production was stimulated by IL-1 beta and TNF-alpha to 450 +/- 155.4% and 531 +/- 170.8% of control, respectively (mean +/- standard error of the mean; P <.05 by analysis of variance), whereas production of inhibin and follistatin was stimulated to a much more modest extent. Similar responses were observed in the choriodecidual explants. Lipopolysaccharide had no significant effect on amnion activin A production, but stimulated choriodecidual production to 290 +/- 34% of control. Lipopolysaccharide exerted only limited effects on inhibin A and follistatin production. CONCLUSIONS: Treatment with proinflammatory mediators resulted in a preferential increase in activin A production compared with that of inhibin A or follistatin. These findings suggest that inflammation of the gestational membranes could result in increased local activin A production and bioactivity. PMID- 11035282 TI - Comparison of active phase labor between triplet, twin, and singleton gestations. AB - OBJECTIVE: To characterize the active phase of labor in triplet pregnancies and compare it with gestational age-matched twins and singletons. METHODS: Active phase rates were calculated beginning at 5 cm of dilation for women with triplet gestations longer than 24 weeks who labored and reached the second stage. Twin and singleton cohorts that also completed the first stage of labor were matched for gestational age at delivery (+/-1 week), parity, and epidural use. Intrapartum variables included oxytocin use (induction or augmentation, duration of infusion, and maximum dosage), cervical dilation at membrane rupture, and active phase dilation rate. RESULTS: Thirty-two triplet pregnancies met inclusion criteria between January 1994 and September 1998 and were each compared with twin and singleton cases in a 1:2 ratio. Triplet and twin active phase rates, while similar (1.8 versus 1.7 cm/hour, respectively), were significantly lower than the mean singleton dilation rate (2.3 cm/hour, P =.02). No other intrapartum variables differed between the three groups. Despite controlling for gestational age at delivery, mean birth weights were significantly higher in singletons and correspondingly lower in twins and triplets (2,493 versus 2,112 and 1,968 g, respectively; P =.001). An analysis of active phase dilation rates as a function of the cumulative birth weight per pregnancy demonstrated an inverse correlation, with slower progress in active labor associated with increasing total fetal weight (R = -.24; P =.002). CONCLUSIONS: Triplet and twin active phase dilation proceeds at a slower rate than that observed in singleton pregnancies. The rate of active phase dilation is inversely correlated to total fetal weight. PMID- 11035283 TI - Periovulatory and interleukin (IL)-1-dependent regulation of IL-6 in the immature rat ovary: a specific IL-1 receptor-mediated eicosanoid-dependent effect. AB - Interleukin (IL)-6, traditionally an IL-1-induced immune regulator, is nevertheless synthesized by a variety of tissues including the ovary. The purposes of this communication were to assess the ovarian expression of IL-6, examine its cyclic variation, and study its regulation by IL-1, a putative intermediary in the ovulatory process. Molecular probing revealed IL-6 transcripts to be most abundant in the thymus, liver, and ovary, which suggests that, in relative terms, untreated immature whole ovarian tissue is a significant site of IL-6 gene expression. Treatment of immature rats with pregnant mare serum gonadotropins resulted in a measurable, statistically significant (P <.05) increase in ovarian IL-6 mRNA compared with untreated controls. Isolated and indeed transient increments in the relative abundance of IL-6 transcripts were noted 4 hours after exposure to hCG, a point in time 8 hours from projected follicular rupture, a pattern highly reminiscent of that previously recognized for IL-1 beta transcripts. Treatment of whole ovarian dispersates from immature rats with IL-1 beta for 48 hours resulted in a significant (P <.05) increase (11 fold) over control in IL-6 transcripts. The IL-1 effect proved dose dependent; the first significant increase was noted at the 1-ng/mL dose level. Evaluation of the time requirements revealed IL-1 beta to significantly (P <.05) up-regulate (4.5-fold) IL-6 transcripts as early as 24 hours into the culture period. Cotreatment with the IL-1 receptor antagonist completely reversed the IL-1 effect, which suggests mediation through a specific IL-1 receptor. Treatment with indomethacin (10 microg/mL), an established inhibitor of prostaglandin biosynthesis, resulted in a significant (P <.05) decrease (79%) in the ability of IL-1 beta to up-regulate IL-6 transcripts. Importantly, the addition of prostaglandin E(2) (10 microg/mL) to untreated or indomethacin-treated cells significantly (P <.05) augmented the IL-1 beta effect. This suggests a role for eicosanoid signaling in IL-1 beta action. Treatment with aminoguanidine, an established inhibitor of nitric oxide synthase, significantly (P <.05) decreased (85%) the IL-1 beta effect. However, the addition of S-nitroso-n-acetyl penicilamine, an established nitrite generator, failed to reverse the aminoguanidine effect, which suggests that the inhibitor effect of aminoguanidine may be nitrite independent. Treatment with cycloheximide produced dose-dependent inhibition of the ability of IL-1 to up-regulate IL-6 transcripts; the maximal inhibitory effect was 89%. Taken together, these findings (1) reaffirm the rat ovary as a site of IL-6 expression; (2) document an in vivo increase in IL-6 transcripts before ovulation; (3) disclose a marked dependence of IL-6 on IL-1 beta; and (4) reveal the IL-1 beta effect to be dose and time dependent, receptor mediated, contingent upon de novo protein biosynthesis, and eicosanoid dependent but nitric oxide independent. These findings suggest that IL-1 action in the ovary may require the intermediacy of IL-6 in a manner like that encountered in extraovarian sites. PMID- 11035284 TI - Changes in serum levels of vascular endothelial growth factor in males and females throughout life. AB - OBJECTIVES: To study serum levels of vascular endothelial growth factor (VEGF), a potent angiogenic factor, during distinct periods of the female life span and compare them with corresponding levels of age-matched males. It is hypothesized that VEGF might be increased at periods of enhanced angiogenesis. METHODS: Venous blood was drawn from healthy females (n = 59) and males (n = 53) divided into six groups: fetuses (cord blood), neonates, children, adults (same females in the proliferative and secretory phases of their menstrual cycle), pregnant, and elderly (postmenopausal). Serum VEGF levels were measured by an enzyme immunoassay. RESULTS: Females showed 49% higher serum VEGF levels than males (t = 2.74, P = .01). Cord and neonatal blood levels were significantly increased compared with those of adults (t = 2.41, P = .02, and t = 5.81, P = .0001, respectively). All female age groups presented higher serum VEGF levels than the group of women in the proliferative phase of the cycle; nevertheless, VEGF levels in the secretory phase did not differ (t = 1.85, P = .07). CONCLUSIONS: Serum VEGF levels are higher in females than in males and during life periods characterized by enhanced growth and development, implying increased rates of angiogenesis. PMID- 11035285 TI - Single mutations of the PTEN gene in recurrent ovarian carcinomas. AB - OBJECTIVE: To establish the role of phosphate and tensin homologue on chromosome 10 (PTEN) mutations in tumorigenesis of the ovary, we determined the mutational spectrum of the PTENgene in surgical specimens of ovarian carcinomas. METHODS: The study group consisted of 86 ovarian cancer specimens (18 fluids, 68 solid specimens), including 30 primary ovarian cancer specimens and 56 of relapsed ovarian cancer from women with a median age of 57.9 years and a range of 27-85 years. Each of the nine exons of the PTEN gene was amplified separately by polymerase chain reaction (PCR). Both strands of the PCR products were sequenced directly by standard cycle sequencing procedures and subsequent computer-aided alignments with the wild-type sequence. RESULTS: In ascitic fluids of two women with recurrence of cancer, we observed mutations: one seven-base-pair insertion at codon 52 (GATGATG) and the other a base-pair substitution resulting in an amino acid change (T131I). We found no mutation in the primary ovarian cancers. CONCLUSIONS: Our data indicate that PTEN mutations have a subordinate role in tumorigenesis of the ovary. PMID- 11035302 TI - [Evolution of surgical treatment for cancer of the larynx in the 20th century]. PMID- 11035303 TI - A limited view of older white men. PMID- 11035304 TI - Health issues unique to the aging man. AB - Several health conditions specifically involve men, including impotence (erectile dysfunction), androgen deficiency syndrome, benign prostatic hypertrophy, prostate cancer, baldness, and gynecomastia. This article reviews the major features of these syndromes and highlights the specific nursing issues involved. PMID- 11035305 TI - Update on osteoporosis in elderly men. AB - Most nurses believe that osteoporosis is a problem of elderly women. Although the disease does occur primarily in women, aging men also are at risk. In fact, osteoporosis in elderly men typically causes greater morbidity and earlier mortality. Currently, no pharmacologic agents have been approved by the Food and Drug Administration to prevent or treat osteoporosis in this population. Several small clinical trials have been conducted in Europe, and several others are in progress in the United States. Geriatric nurses need to be aware of the risk of osteoporosis in elderly men and provide early counseling and prevention strategies. PMID- 11035306 TI - The controversy of prostate screening. AB - Within the context of aging and ageism, this article evaluates current research on the benefits and risks of prostate cancer screening. Prostate cancer is a concern for many men because it often strikes in the prime of life. Prostate cancer is the most common form of cancer in American men and the second leading cause of cancer-related mortality after lung cancer. Although the disease can be cured if discovered early, it is a slow-growing malignancy that leads to an agonizing death if left untreated. Differential diagnosis often is complicated by comorbid conditions that are part of the normal aging process. Moreover, practitioners' judgment often is prejudiced by common aging misconceptions. Despite the benefits of early screening and detection, the benefits of screening initiatives remain controversial. PMID- 11035307 TI - Exploring the efficacy of support groups for men with prostrate cancer. AB - For the 180,000 men who will be diagnosed with prostate cancer this year, improvements in the disease's medical management have prolonged life expectancy. Long-standing treatment side effects include bowel and urinary incontinence and erectile dysfunction, which frequently are embarrassing and contribute to poor emotional well-being. Despite the popularity of support groups in alleviating emotional symptoms related to cancer and its treatment, few men participate in such groups. This article focuses on the reasons why men do not find support groups appealing and presents alternatives that may be more acceptable. Suggestions are made for clinical and research implications. PMID- 11035308 TI - The forgotten faces: the lonely journey of powerlessness experienced by elderly single Chinese men with heart disease in Taiwan. AB - The purpose of this study is to explore the content and background context of powerlessness experienced by elderly single Chinese men with heart disease and their coping behaviors during their hospitalization stage. Data were gathered by semi-structured interviews at a leading veterans' hospital in northern Taiwan and analyzed using a qualitative content analysis mode. Twenty-six men who were 65 or older, diagnosed with heart disease, and who lived alone during the preadmission stage were recruited. Eighty-one percent (N = 21) reported that their perceptions of powerlessness occurred either in the preadmission or hospitalization stage or were expected to occur after discharge. Other complaints of powerlessness were attributed to having no choices in appropriate living places during the preadmission stage, having no control over discomfort, being unable to obtain care and companionship from families and friends, failing to get medical information about their disease and options of treatment during hospitalization, or expecting deteriorating health and receiving no assistance during emergencies or in the dying stage after discharge from the hospital. PMID- 11035309 TI - Graham McDougall, Jr., PhD, RN, CS: Academic, Leader, and Researcher. PMID- 11035310 TI - A framework for nurse staffing in long-term care facilities. AB - Nurse staffing is complex and requires the nurse administrator to consider multiple factors. The Framework for Nurse Staffing in Long-term Care (LTC) Facilities provides a comprehensive perspective of those factors and their interrelatedness. The purpose of the framework is to provide nurse administrators and managers with ways to analyze and evaluate nurse staffing in their own facilities and develop solutions and approaches that are specific to their needs and circumstances. The framework also can serve as the basis for developing and testing research questions that will guide nurse administrators in making informed decisions to determine, allocate, and deliver the resources necessary to provide quality care for residents. PMID- 11035311 TI - Correlates and consequences of chronic pain in older adults. AB - Chronic pain is a frequent challenge to older adults' coping skills. Despite the widespread occurrence of chronic geriatric pain, no comprehensive body of literature on this topic exists. Instead, research on chronic pain is scattered across disciplines and is perceived as inaccessible by scientists. We completed a comprehensive review and qualitative analysis of the geriatric chronic pain literature since 1990 and found 314 articles on this topic that reported. North American research. Physical, social, and psychologic variables associated with chronic pain and the elderly were mentioned in just over half (53%) of the articles. However, both psychosocial causes and consequences of chronic pain were understudied. Only 16% of the articles had social variables (gender, race, and age) as their primary interest; 27% focused on psychologic or psychiatric issues, with half including depression as the variable of interest. An analysis of the articles' content suggests that research on chronic pain in later life would be substantially improved if a more structured and comprehensive approach were used that combined the study of psychosocial issues with that of physical pain. Researchers and clinicians with a global understanding of chronic pain might help improve quality of life for older adults. PMID- 11035312 TI - Report from the World Alzheimer Congress 2000. PMID- 11035313 TI - Resources for Home Care Nurses. PMID- 11035314 TI - Should the definition of preeclampsia include a rise in diastolic blood pressure of >/=15 mm Hg to a level <90 mm Hg in association with proteinuria? AB - OBJECTIVE: This study was undertaken to compare baseline characteristics and pregnancy outcomes between normotensive women who did and those who did not have a rise in diastolic blood pressure of >/=15 mm Hg in association with proteinuria. STUDY DESIGN: We studied 4302 healthy nulliparous women from the Calcium for Preeclampsia Prevention trial who were delivered at >/=20 weeks' gestation. We selected as the study group normotensive women who developed proteinuria within 7 days of a rise in diastolic blood pressure of >/=15 mm Hg with respect to baseline on 2 occasions 4 to 168 hours apart. Baseline blood pressure was the mean of measurements at 2 clinic visits before 22 weeks' gestation. Other normotensive women used for comparison were those who did not develop gestational hypertension or a rise in diastolic blood pressure of >/=15 mm Hg in association with proteinuria. RESULTS: Except for greater weight (P <.001), body mass index (P <.001), and systolic blood pressure (P =.05) the baseline characteristics of the 82 women with a rise in diastolic blood pressure of >/=15 mm Hg in association with proteinuria did not differ significantly from those of the other normotensive women. Although they had a greater rate of weight gain (P <.005), larger babies (P =.06), and a 2-fold increase in abdominal delivery (P <.001), there was little other evidence of adverse pregnancy outcomes among these women. CONCLUSION: During normotensive pregnancy a rise in diastolic blood pressure of >/=15 mm Hg in association with proteinuria appears to be benign and is not a useful clinical construct. PMID- 11035315 TI - Magnesium prevents seizure-induced reduction in excitatory amino acid receptor (kainate and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid) binding in pregnant rat brain. AB - OBJECTIVE: Our purpose was to evaluate the effect of seizures on kainate and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor binding in maternal rat brain and whether maternal peripheral administration of magnesium sulfate can decrease this effect. STUDY DESIGN: Rats were implanted with a bipolar electrode into the hippocampus. One week of recovery was allowed before breeding. Pregnant rats were randomly assigned to 1 of 4 groups, as follows: group 1, sodium chloride and no seizures (n = 5); group 2, magnesium sulfate and no seizures (n = 4); group 3, sodium chloride and seizures (n = 8); and group 4, magnesium sulfate and seizures (n = 9). Doses of sodium chloride or magnesium sulfate were administered every 20 minutes for 4 hours to all rats on gestational days 9, 11, 13, 15, 17, and 19, followed by seizure induction (groups 3 and 4). On gestational day 20, rats were perfused, brains were dissected, and cryostat sections were taken, labeled in vitro, and placed on Hyperfilm for 4 weeks. The ligands used included kainate receptor agonist and alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid receptor agonist and antagonist. Optical density measurements of binding in 15 brain regions on each section were evaluated by 1- and 2-way analysis of variance. RESULTS: Seizure activity was associated with decreased alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor binding of its agonist in pregnant rat brains (seizure effect, 25.9 +/- 3.2 and 92.6 +/- 3.4 fmol/mg tissue in hindbrain and forebrain, respectively; no seizure effect, 44.5 +/- 4.7 and 110. 7 +/- 5.0 fmol/mg tissue in hindbrain and forebrain, respectively; P <.01). Magnesium administration was associated with increased binding of tritiated alpha-amino-3-hydroxy-5-methylisoxazole-4 propionic acid (magnesium effect, 44.9 +/- 4.2 and 110.4 +/- 4.5 fmol/mg tissue in hindbrain and forebrain, respectively; sodium chloride effect, 25.5 +/- 3.7 and 92.9 +/- 4.0 fmol/mg tissue in hindbrain and forebrain, respectively; P <.01). The same trend was seen with the kainate receptor in the hippocampus and hypothalamus, with a significant interaction effect between seizure and magnesium (P <.05). CONCLUSIONS: The mechanism for maternal rat brain injury resulting from seizure activity may be, at least in part, associated with alteration in the function of excitatory amino acid receptors. Administration of magnesium sulfate can counteract this effect and may reduce resultant maternal brain damage. PMID- 11035316 TI - Effects of thrombin on myometrial contractions in vitro and in vivo. AB - OBJECTIVE: A previous report from our laboratory demonstrated that thrombin stimulates myometrial contractions by activating the phosphatidylinositol signaling pathway in a dose-dependent fashion. The studies described in this report sought to determine whether thrombin and blood stimulate myometrial contractions both in vivo and in vitro and whether these uterotonic effects could be suppressed or prevented with thrombin inhibitors. STUDY DESIGN: In vitro contraction studies were performed with proestrus and estrus rat myometrial tissue. In vivo contraction studies were performed with nonpregnant and timed pregnant Sprague-Dawley rats. RESULTS: Pretreatment of thrombin with hirudin suppressed the uterotonic effects of thrombin in vitro. Fresh whole blood stimulated myometrial contractions in a dose-dependent fashion in vitro, and thrombin inhibitors decreased the myometrial response seen with blood alone. Thrombin increased the frequency, intensity, and tone of myometrial contractions in vivo in a dose-related manner. In pregnant animals increasing doses of whole blood increased the frequency and tone of myometrial contractions. In both pregnant and nonpregnant animals whole blood significantly stimulated myometrial contractions, whereas heparinization of the blood significantly suppressed this in vivo uterotonic effect. CONCLUSION: Thrombin is a potent uterotonic agent both in vitro and in vivo; furthermore, the uterotonic effects of blood appeared to be related to thrombin production during coagulation. These studies provide a possible mechanistic explanation for the observed increase in myometrial contractions in the presence of intrauterine bleeding and may also provide an insight into preterm labor associated with vaginal bleeding. PMID- 11035317 TI - Second-trimester plasma homocysteine levels and pregnancy-induced hypertension, preeclampsia, and intrauterine growth restriction. AB - OBJECTIVE: The purpose of this study was to determine whether second-trimester plasma homocysteine levels are elevated among women whose pregnancies are subsequently complicated by pregnancy-induced hypertension, preeclampsia, or intrauterine growth restriction. STUDY DESIGN: Women with normal but relatively low plasma zinc levels were randomly assigned to receive zinc supplementation or placebo from 19 weeks' gestation until delivery. Plasma homocysteine concentration and plasma and erythrocyte folate levels were determined for all available stored samples (zinc group, 231/294; placebo group, 206/286) at 26 and 37 weeks' gestation. Among all women with available samples, pregnancy-induced hypertension (n = 12) or preeclampsia (n = 4) developed in 16 women, and 22 pregnancies were complicated by intrauterine growth restriction. RESULTS: Mean homocysteine levels in women with pregnancy-induced hypertension and preeclampsia were similar to those of control subjects at 26 weeks' gestation but were significantly higher at 37 weeks' gestation. Homocysteine levels were similar between women with pregnancies complicated by intrauterine growth restriction and control subjects at both time points. CONCLUSION: Second-trimester plasma homocysteine concentrations do not predict the subsequent development of pregnancy-induced hypertension, preeclampsia, and intrauterine growth restriction. PMID- 11035318 TI - Neonatal sepsis and death after multiple courses of antenatal betamethasone therapy. AB - OBJECTIVE: This study was undertaken to compare the effects of single versus multiple courses of betamethasone therapy on the frequencies of neonatal outcomes and perinatal infectious morbidity among singleton pregnancies complicated by preterm delivery. STUDY DESIGN: We performed a nonconcurrent prospective analysis of singleton pregnancies delivered between 24 and 34 weeks' gestation after antenatal betamethasone exposure. Patients were categorized into two groups according to betamethasone exposure: (1) two 12-mg doses in a 24-hour interval on admission (single-course group) and (2) repeated dosing after the initial single course (multiple-course group). All patients received prophylactic antibiotics for group B streptococci. Any patients with ruptured membranes for >24 hours before delivery were excluded. Data were analyzed with the Student t test, the chi(2) test, and the Fisher exact test. Multiple logistic regression analyses were performed to examine the effect of each steroid dosing regimen on early onset neonatal sepsis and neonatal death. P <.05 was considered significant for all 2-tailed tests. RESULTS: A total of 453 patients were included, with 267 in the single-course group and 186 in the multiple-course group. The two groups were similar with respect to maternal demographic characteristics, gestational age at delivery, mode of delivery, birth weight, and maternal group B streptococcal colonization. Multiple courses were significantly associated with early-onset neonatal sepsis (odds ratio, 5.00; 95% confidence interval, 1.3-23. 2), chorioamnionitis (odds ratio, 9.96; 95% confidence interval, 2. 1-64.6), endometritis (odds ratio, 3.61; 95% confidence interval, 1. 7-8.1), and neonatal death (odds ratio, 2.92; 95% confidence interval, 1.3-6.9). The frequencies of the other neonatal outcomes analyzed, including respiratory distress syndrome and grade III or IV intraventricular hemorrhage, were similar between the 2 groups. Multiple logistic regression analyses confirmed that multiple courses of antenatal betamethasone were independently associated with early-onset neonatal sepsis (odds ratio, 1.25; 95% confidence interval, 1.1-1.9) and neonatal death (odds ratio, 1.70; 95% confidence interval, 1.1-1.9). CONCLUSIONS: Multiple courses of antenatal betamethasone are associated with increased risks of perinatal infectious morbidity and neonatal death. PMID- 11035319 TI - Mathematic modeling to predict abruptio placentae. AB - OBJECTIVE: This study was undertaken to identify correlates of abruptio placentae and to develop a mathematic model for the prediction of abruptio placentae. STUDY DESIGN: A total of 170,258 singleton birth records from 1991 to 1996 contained in the Schleswig-Holstein perinatal database were analyzed. Fifty-two recognized obstetric risk factors were subjected to univariate analysis. Correlates of abruptio placentae then underwent stepwise forward binary logistic regression. A constant value B(0), coefficients B(1) through B(p), an odds ratio, and a 95% confidence interval were calculated for individual correlates. RESULTS: Abruptio placentae occurred in 874 of 170,258 singleton gestations (0.5%). Of the 52 risk factors 31 proved to be correlates of abruptio placentae, with 16 among primiparous women and 25 among multiparous women. Ten correlates for primiparous, women and 13 for multiparous women emerged from the linear regression, with 7 correlates being shared by both primiparous and multiparous women. CONCLUSION: The probability that abruptio placentae will occur (p) can be calculated according to the following expression: p = e (z)/(1 + e (z)), where z = B(0) + B(1), em leaderB(p). For example, for a primiparous woman who smokes with bleeding at >28 weeks' gestation and a male fetus in the breech position, the following calculation would yield the chance of abruptio placentae:z = -2.25 + 2.51 + 0.41 + 0.24 + 0.60 = 1.51; p = e (1.51)/ (1 + e (1.51)) = 4. 53/5.53 = 0.82, or 82%. PMID- 11035320 TI - Cervical incompetence prevention randomized cerclage trial (CIPRACT): study design and preliminary results. AB - OBJECTIVE: The objective of this study was to compare different management strategies for women at risk for cervical incompetence. STUDY DESIGN: In an ongoing randomized trial patients with a previous preterm delivery at <34 weeks' gestation who met clinical criteria for the diagnosis of cervical incompetence are allocated to receive a prophylactic cerclage (prophylactic cerclage group) or not (observational group) in a proportion of 1:2. Transvaginal ultrasonographic follow-up examination of the cervix is performed in both groups. When a patient of the latter group has a cervical length <25 mm at <27 weeks' gestation, a further random assignment of therapeutic cerclage or no cerclage is performed. The analysis is by intent to treat. RESULTS: Primary random assignment allocated 23 women to the prophylactic cerclage group and 44 to the observational group. Both groups were comparable with respect to obstetric history. No significant difference was found between the prophylactic cerclage group and the observational group in preterm delivery at <34 weeks' gestation (3/23 vs 6/44, respectively) and neonatal survival (21/23 vs 41/44, respectively). A cervical length <25 mm was found in 18 patients (41%) in the observational group at a mean gestational age of 19.1 +/- 2.9 weeks' gestation. Incidence of preterm delivery at <34 weeks' gestation was significantly higher in the group with short cervical length (6/18 vs 0/26; P =.003). Secondary random assignment of the 18 patients with short cervical length allocated 10 to undergo therapeutic cerclage. Preterm delivery at <34 weeks' gestation was significantly less frequent in the therapeutic cerclage group (1/10 vs 5/8). CONCLUSION: Transvaginal ultrasonographic serial follow-up examinations of the cervix in women at risk for cervical incompetence, with secondary intervention as indicated, appears to be a safe alternative to the traditional prophylactic cerclage. Transvaginal ultrasonographic follow-up examination of the cervix can save the majority of women from unnecessary intervention. Placement of a therapeutic cerclage may reduce the incidence of preterm delivery at <34 weeks' gestation among high-risk patients. PMID- 11035321 TI - A randomized trial of cerclage versus no cerclage among patients with ultrasonographically detected second-trimester preterm dilatation of the internal os. AB - OBJECTIVE: The aim of this study was to compare perinatal outcomes of patients with second-trimester ultrasonographic evidence of preterm dilatation of the internal os treated with cerclage versus those of patients not treated with cerclage. STUDY DESIGN: From May 1998 through June 1999 patients with ultrasonographic evidence of preterm dilatation of the internal os between 16 and 24 weeks' gestation were randomly assigned to receive a McDonald cerclage or no cerclage. Before random assignment all patients underwent amniocentesis and urogenital cultures and then received 48 hours of therapy with indomethacin and antibiotics. After treatment each patient was followed up as an outpatient with bed rest and weekly ultrasonographic evaluation. RESULTS: Of the 61 patients 31 were randomly assigned to cerclage and 30 were randomly assigned to no cerclage. There were no differences between groups with respect to maternal demographic characteristics, risk factors for preterm birth, cervical measurements, rescue procedures, readmission, chorioamnionitis, and abruptio placentae. The mean gestational age at delivery (33.5 +/- 6.3 weeks) and the perinatal death rate (12. 9%) in the cerclage group were similar to the mean gestational age at delivery (34.7 +/- 4.7 weeks; P =.4) and the perinatal death rate (10.0%; P =.9) in the no-cerclage group. CONCLUSION: Treatment with McDonald cerclage of preterm dilatation of the cervix detected ultrasonographically during the second trimester did not improve perinatal outcomes. PMID- 11035322 TI - Patients with a prior failed transvaginal cerclage: a comparison of obstetric outcomes with either transabdominal or transvaginal cerclage. AB - OBJECTIVE: Our purpose was to compare the incidence of preterm birth after a prior failed vaginal cerclage in patients who had a subsequent transabdominal or a transvaginal cerclage. STUDY DESIGN: We conducted a retrospective cohort study of singleton pregnancies in women who had undergone (9-14 weeks) either a transabdominal or a transvaginal prophylactic cerclage after >/=1 prior failed transvaginal cerclage. Prior failed transvaginal cerclage was defined as a preterm birth at <33 weeks' gestation in the immediate prior pregnancy despite a transvaginal cerclage. All transabdominal cerclage procedures were performed by a single attending physician (George Davis, DO). Patients with a cervix too short for transvaginal cerclage placement, placenta previa, or major fetal anomalies were excluded. Primary outcome was preterm birth at <35 weeks' gestation. RESULTS: Forty transabdominal and 24 transvaginal cerclage pregnancies were analyzed. These 2 groups were similar in race and payer status but differed in age (34.0 +/- 4.2 vs 31.3 +/- 4.6 years, respectively; P =.01). The transabdominal cerclage group had more prior failed cerclage procedures per patient (1.8 +/- 1.0 vs 1.1 +/- 0.3; P =.02) and more prior 14- to 24-week spontaneous abortions per patient (2.4 +/- 1.3 vs 1.5 +/- 1.0; P =.02) than the transvaginal cerclage group. Preterm delivery at both <35 and <33 weeks' gestation was less common in the transabdominal cerclage group (18% vs 42%, P =.04; 10% vs 38%, P =.01; respectively) than in the transvaginal cerclage group. Gestational age at delivery was 36. 3 +/- 4.1 weeks in the transabdominal cerclage group and 32.8 +/- 8. 6 weeks in the transvaginal cerclage group (P =.03). Preterm premature rupture of membranes also occurred less often in the transabdominal cerclage group than in the transvaginal cerclage group (8% vs 29%, P =.03). CONCLUSION: In patients with a prior failed transvaginal cerclage, transabdominal cerclage is associated with a lower incidence of preterm delivery and preterm premature rupture of membranes in comparison with transvaginal cerclage. PMID- 11035323 TI - Management of cervical cerclage and preterm premature rupture of the membranes: should the stitch be removed? AB - OBJECTIVE: Our aim was to determine whether retention of cerclage after preterm premature rupture of the membranes occurring before 34 completed weeks' gestation influences pregnancy outcome. STUDY DESIGN: Singleton pregnancies with cerclage and premature rupture of the membranes between 24.0 and 34.9 weeks were reviewed. Women were excluded if they were first seen in labor, had chorioamnionitis, or were delivered within 48 hours. Control subjects consisted of women with premature rupture of the membranes without cerclage. RESULTS: Eighty-one cases of cerclage with premature rupture of the membranes met criteria for inclusion: 30 women (37%) had their cerclage removed at presentation, and 51 (63%) retained the cerclage until delivery. Cases were similar in terms of gestational age at placement and gestational age at premature rupture of the membranes. There was no significant difference between the retained, removed, or control groups in terms of latency, gestational age at delivery, chorioamnionitis, or neonatal morbidity and mortality. CONCLUSIONS: Retention of cervical cerclage after premature rupture of the membranes occurring before 34 completed weeks' gestation is associated with comparable clinical outcomes with respect to latency and perinatal outcome, when compared with removal of the cerclage. PMID- 11035324 TI - Timing of cerclage removal after preterm premature rupture of membranes: maternal and neonatal outcomes. AB - OBJECTIVE: Our aim was to evaluate immediate versus delayed removal of cerclage for women with preterm premature rupture of membranes with respect to maternal and neonatal outcomes. STUDY DESIGN: We retrospectively analyzed women with preterm premature rupture of membranes at <34 weeks' gestation with prior cerclage placement. Exclusion criteria included presentation with chorioamnionitis, active labor, or nonreassuring fetal status. Timing of cerclage removal, immediate (<24 hours) or delayed (>24 hours), was compared. RESULTS: There were 25 women in the delayed-removal group and 37 in the immediate-removal group. Average times to removal were 206.8 +/- 7.4 and 5.4 +/- 0.2 hours, respectively. Use of betamethasone was similar for both groups; however, antenatal antibiotic use (100% vs 80%; P =.03) and short-term tocolytic use (20% vs 3%; P =.04) were higher in the delayed-removal group. Duration of latency was significantly longer with delayed removal (10.1 vs 5.0 days; P <. 001). Delivery occurred >48 hours from preterm premature rupture of membranes in 96% (24/25) versus 54% (20/37; P <.001) and >7 days from rupture in 56% (14/25) versus 24% (9/37; P =.02), respectively. Rates of neonatal sepsis (at <10 days) and maternal infection were not statistically different. Neonatal outcomes did not significantly differ regarding mortality, respiratory distress syndrome, birth weight, or duration of stay in the intensive care nursery. CONCLUSION: With the current management scheme for preterm premature rupture of membranes, cerclage retention significantly increases duration of latency without significantly altering maternal or neonatal outcomes. PMID- 11035325 TI - Expectant management of severe preterm preeclampsia: is intrauterine growth restriction an indication for immediate delivery? AB - OBJECTIVE: Expectant management of severe preterm preeclampsia is gaining widespread acceptance in clinical practice. The objective of our study was 2-fold to determine the frequency of fetal deterioration with expectant management of severe preterm preeclampsia and to evaluate whether the presence of intrauterine growth restriction on admission is associated with a shorter admission-to delivery interval or more deliveries resulting from nonreassuring fetal status in comparison with pregnancies with preeclampsia but without intrauterine growth restriction. STUDY DESIGN: This was an observational study of women with singleton pregnancies at <34 completed weeks' gestation who were admitted to the hospital with the diagnosis of severe preeclampsia and managed expectantly. Fetal status on admission, admission-to-delivery interval, indication for delivery, and neonatal outcome were examined. RESULTS: Forty-seven women were studied during a 3-year period (1996-1999). Gestational age at admission was 29.8 +/- 2.6 weeks. The mean admission-to-delivery interval for the entire group was 6.0 +/- 5.1 days; in 42.5% delivery was for fetal indications. In comparison with the absence of intrauterine growth restriction, the presence of intrauterine growth restriction at admission resulted in a significantly shorter admission-to delivery interval (3.1 +/- 2.1 vs 6.6 +/- 6.1 days; P <.05). Most fetuses with intrauterine growth restriction (85.7%) were delivered before 1 week. Although 57% of fetuses with intrauterine growth restriction were delivered for fetal indications, versus 39% of fetuses without intrauterine growth restriction, these rates were not found to be significantly different. Neonatal outcomes, as reflected by Apgar scores, number of admissions to and duration of stay in the neonatal intensive care unit, and neonatal mortality rates, were similar. CONCLUSION: Pregnancies complicated by severe preterm preeclampsia and the presence of intrauterine growth restriction at admission may not benefit from expectant management beyond the 48 hours needed for betamethasone to act. Furthermore, all patients may benefit from close fetal monitoring before delivery because of the high rate of intervention for deteriorating fetal status. PMID- 11035326 TI - Group B streptococci causing neonatal bloodstream infection: antimicrobial susceptibility and serotyping results from SENTRY centers in the Western Hemisphere. AB - OBJECTIVE: Group B streptococcal infection is a common cause of neonatal sepsis. Surveillance of antimicrobial susceptibility and serotype frequencies of invasive group B streptococci is important to ensure the effectiveness of therapeutic regimens and to guide vaccine development. STUDY DESIGN: Prospective surveillance of neonatal bloodstream infection was performed at all Western Hemisphere sites participating in the SENTRY Program. From January 1997 through December 1999, a total of 122 isolates of bloodstream infections with group B streptococci were collected and sent to the University of Iowa for antimicrobial susceptibility testing and serotyping. RESULTS: No isolates were resistant to penicillin. More than 25% of isolates from US hospitals and 14% of isolates from Canadian hospitals were erythromycin resistant. Seven percent of isolates from the United States and Canada were resistant to clindamycin. No clindamycin or erythromycin resistance was found among isolates from Latin America. Clindamycin and erythromycin resistance was most frequent among serotype V strains. CONCLUSIONS: No emerging resistance to penicillin was noted among bloodstream infection isolates of group B streptococci from a broad geographic area; erythromycin and clindamycin resistance was found in the United States and Canada and appeared most frequently among serotype V strains. PMID- 11035327 TI - Hypoperfusion causes increased production of interleukin 6 and tumor necrosis factor alpha in the isolated, dually perfused placental cotyledon. AB - OBJECTIVE: Our purpose was to determine whether exposure of the isolated, perfused human placental cotyledon to different fetal circuit perfusion rates, and to concomitant pressure differences, alters placental production of interleukin 6 and tumor necrosis factor alpha. STUDY DESIGN: The maternal and fetal circulations of 2 cotyledons from 5 placentas were perfused for 4 hours. The fetal circulation of 1 cotyledon was perfused at a low rate of 1 mL/min, and the other at a high rate of 10 mL/min. The maternal circulation of each cotyledon was perfused at 10 mL/min. Effluents from the fetal circulation were collected at hourly intervals, and concentrations of interleukin 6 and tumor necrosis factor alpha were determined by enzyme-linked immunosorbent assay. Concentrations of interleukin 6, obtained from a prior study with an estimated physiologic fetal circulation rate of 4 mL/min, were compared with the low and high perfusion rate results. RESULTS: Concentrations of interleukin 6 and tumor necrosis factor alpha were greater at the perfusion rate of 1 mL/min, in comparison with the perfusion rate of 10 mL/min, with statistically significant differences achieved at 2 and 4 hours for interleukin 6 and at 4 hours for tumor necrosis factor alpha. Concentrations of both cytokines increased exponentially with time. Placental perfusion pressures were significantly greater at the perfusion rate of 10 mL/min. CONCLUSION: Placental hypoperfusion results in an increased production of both interleukin 6 and tumor necrosis factor alpha. This finding links placental perfusion abnormalities to the myriad of disorders associated with elevated concentrations of inflammatory cytokines, including cerebral palsy. PMID- 11035328 TI - Interleukin 6 determinations in cervical fluid have diagnostic and prognostic value in preterm premature rupture of membranes. AB - OBJECTIVE: Our aim was to determine whether interleukin-6 concentrations in cervical fluid samples are of value in the identification of microbial invasion of the amniotic cavity, prediction of the duration of the latency period, and assessment of the risk of neonatal complications in preterm premature rupture of membranes. STUDY DESIGN: A cohort study was performed in 86 patients with preterm premature rupture of membranes. Amniotic fluid and cervical fluid were collected. Amniotic fluid was cultured for aerobic and anaerobic bacteria, as well as mycoplasmas. Interleukin 6 was measured by a sensitive and specific immunoassay. The receiver operating characteristic curve, logistic regression, and survival techniques were used for analysis. RESULTS: (1) Patients with a positive amniotic fluid culture had a significantly higher median cervical fluid interleukin 6 concentration than those with negative results (median, 528 pg/mL; range, 174-825 pg/mL; vs median, 169 pg/mL; range, 8-986 pg/mL; P <.0001). (2) A cervical fluid interleukin 6 concentration of >350 pg/mL had a sensitivity of 92% and a specificity of 78% in the identification of a positive amniotic fluid culture. (3) Patients with a cervical fluid interleukin 6 concentration of >350 pg/mL had a significantly shorter median interval to delivery and higher rate of funisitis, preterm delivery within 2 days and 7 days, and the occurrence of significant neonatal morbidity than did those with a cervical fluid interleukin 6 concentration of <350 pg/mL (P <.05 for each). (4) The increased perinatal morbidity remained significant after adjustment for gestational age (P <.05). (5) There was a strong correlation between cervical fluid concentrations and amniotic fluid concentrations of interleukin 6 (P <.001). CONCLUSION: Cervical fluid interleukin 6 determinations are of value in the assessment of the likelihood of microbial invasion of the amniotic cavity, impending preterm delivery, and the occurrence of significant neonatal complications in the setting of preterm premature rupture of membranes. PMID- 11035329 TI - Plasma, urinary, and salivary 8-epi-prostaglandin f2alpha levels in normotensive and preeclamptic pregnancies. AB - OBJECTIVE: Our purpose was to measure and compare plasma, urinary, and salivary concentrations of 8-epi-prostaglandin F(2alpha) (8-isoprostane) in women with normotensive pregnancies and the respective concentrations in pregnancies complicated by preeclampsia. STUDY DESIGN: Plasma, urinary, and salivary 8 isoprostane levels were measured in pregnant women with preeclampsia (n = 40), normotensive pregnant women (n = 20), and nonpregnant women (n = 10). One-way analysis of variance was used to determine significant differences. RESULTS: Plasma free 8-isoprostane concentrations were increased in women with severe preeclampsia (342 +/- 50 pg/mL), in comparison with nonpregnant women (129 +/- 17 pg/mL) and normotensive pregnant women (150 +/- 11 pg/mL; P =.003, and.0001, respectively). Urinary excretion of 8-isoprostane was slightly but not significantly decreased in preeclampsia (1200 +/- 227 pg/mL), in comparison with urinary excretion in nonpregnant women (1625 +/- 364 pg/mL) and normotensive pregnant women (2149 +/- 432 pg/mL). Salivary concentrations of 8-isoprostane were increased in normotensive women (496 +/- 113 pg/mL), in comparison with nonpregnant women (150 +/- 27 pg/mL) but were not related to preeclampsia (419 +/ 96 pg/mL; P 4 days between January 1991 and June 1998 were included. Amniotic fluid volume was assessed as the maximum cord-free pocket with serial ultrasonographic examinations. Consenting women with persistent (>4 days) oligohydramnios (amniotic fluid 2 cm. The pregnancy, neonatal, and long-term neurologic outcomes of the cases that spontaneously maintained a median amniotic fluid pocket >2 cm (amnioinfusion-not-necessary group) were compared with those of women with oligohydramnios who underwent amnioinfusion but continued to have a median amniotic fluid pocket after preterm premature rupture of membranes 2 cm for >/=48 hours in 11 (30%) patients. This successful amnioinfusion group was comparable with the persistent oligohydramnios group (n = 25) in gestational age at first amnioinfusion (median, 20.2 weeks; range, 16-25.6 weeks; vs median, 20.3 weeks; range, 16.5-24.2 weeks; P =.4), number of amnioinfusions (median, 3; range, 1-9; vs median, 3; range, 1-5; P =.4), and interval between amnioinfusions (median, 6 days; range, 4-14 days; vs median, 8 days; range, 6-43 days; P =. 1). However, patients in the persistent oligohydramnios group had a significantly shorter interval to delivery, lower neonatal survival (20%), and higher rates of pulmonary hypoplasia (62%) and abnormal neurologic outcomes (60%) than the patients in the groups in which amnioinfusion was not necessary or was successful (all P 7 days from dosing and those of matched untreated infants. STUDY DESIGN: Eighteen infants who received multiple courses of corticosteroids and were delivered within 7 days of dosing were matched with 18 infants who received 1 course of corticosteroids >7 days before delivery (remote) and 18 untreated infants. Respiratory compliance and functional residual capacity were measured within 36 hours. Differences were compared by analysis of variance. RESULTS: Infant demographics were similar. Respiratory compliance was higher in the multiple-course group than in the remote or untreated group (P <.02). Functional residual capacity was higher in the multiple-course group than in the untreated group (P <.05) but similar to that found in the remote group. CONCLUSION: Babies delivered after multiple courses of corticosteroids and within 7 days of dosing demonstrated improved respiratory compliance compared with untreated and remotely treated infants. This suggests that the enzyme system responsible for surfactant production can be repetitively induced despite prior treatment with corticosteroids. The increased functional residual capacity in remotely treated infants may reflect a maturation of lung architecture independent of surfactant production. PMID- 11035334 TI - End-tidal carbon monoxide measurements in women with pregnancy-induced hypertension and preeclampsia. AB - OBJECTIVE: We sought to compare the end-tidal carbon monoxide breath levels in pregnant women with and without pregnancy-induced hypertension and preeclampsia. STUDY DESIGN: We prospectively performed end-tidal carbon monoxide measurements corrected for ambient carbon monoxide in nonsmoking women during late gestation (>31 weeks). The study group included 22 women with pregnancy-induced hypertension or symptoms of preeclampsia and a control group of 20 normotensive pregnant women. RESULTS: The carbon monoxide measurements corrected for ambient carbon monoxide (mean +/- SD) were significantly lower (P <.01) in the hypertensive group than in the control group (1.17 +/- 0.35 vs 1.70 +/- 0.54 ppm). The study group had a significantly higher number of low (<1.2 ppm) end tidal carbon monoxide measurements corrected for ambient carbon monoxide (13 [59.1%] vs 1 [5.0%]; P <.001). The end-tidal carbon monoxide measurements corrected for ambient carbon monoxide remained significantly lower in comparison with those found in the control group when the study group was divided into women with pregnancy-induced hypertension only (n = 11) and those with preeclampsia (n = 11) (1.19 +/- 0.37 ppm; P <.01; and 1.15 +/- 0.41 ppm; P <.01; respectively). CONCLUSIONS: Our findings suggest that carbon monoxide formation may be significantly lower in women with pregnancy-induced hypertension and preeclampsia. These data suggest that carbon monoxide could have a contributory role in the apparent paradox of the seemingly protective effect of smoking to decrease the risk of preeclampsia. PMID- 11035335 TI - Lactoferrin in intrauterine infection, human parturition, and rupture of fetal membranes. AB - OBJECTIVE: Lactoferrin is an iron-binding protein with antimicrobial properties. This study was undertaken to determine whether amniotic fluid concentrations of this protein change with gestational age, infection, labor, and rupture of membranes. STUDY DESIGN: This cross-sectional study included women who underwent transabdominal amniocentesis (n = 268) in the following groups: (1) mid trimester of pregnancy; (2) preterm labor who delivered at term, preterm labor who delivered preterm with intra-amniotic infection, and preterm labor who delivered preterm without intra-amniotic infection; (3) preterm premature rupture of membranes in the presence or absence of intra-amniotic infection; (4) term with intact membranes not in labor, in labor, and in labor with intra-amniotic infection; and (5) premature rupture of membranes at term not in labor. In addition, lactoferrin concentrations were determined in maternal plasma and cord blood of patients at term not in labor. Lactoferrin concentration was measured with an immunoassay. RESULTS: (1) Lactoferrin was detectable in 85.4% (229/268) of amniotic fluid samples, not detectable in all fluid obtained in the mid trimester, and detectable in all maternal and cord plasma samples. (2) The concentration of lactoferrin increased with advancing gestational age (r = 0.68; P <.0001). (3) Intra-amniotic infection was associated with significant increases in amniotic fluid lactoferrin concentrations in patients with preterm labor (no intra-amniotic infection median, 1641.2 ng/mL; range, <1.24-35,090.0 ng/mL; vs intra-amniotic infection median, 3833.6 ng/mL; range, 746.0-47,020.0 ng/mL; P <.001), term labor (no intra-amniotic infection median, 2085.8 ng/mL; range, 425.0-23,230.0 ng/mL; vs intra-amniotic infection median, 5627.0 ng/mL; range, <1.24-19,220.0 ng/mL; P <. 001), and preterm premature rupture of membranes (no intra-amniotic infection median, 2190 ng/mL; range, <1.24-7456.1 ng/mL; vs intra amniotic infection median, 3449.3 ng/mL; range, <1.24-83,600. 0; P <.01). (4) Spontaneous labor at term but not preterm was associated with a significant decrease in amniotic fluid lactoferrin concentration (P <.05). (5) Spontaneous term parturition was associated with a significant increase in umbilical cord plasma lactoferrin concentration (P <.005). CONCLUSION: (1) Intra-amniotic infection was consistently associated with dramatically increased concentrations of lactoferrin in amniotic fluid. (2) Term parturition was associated with a significant increase in lactoferrin concentration in the fetal compartment (umbilical cord blood) and a decrease in the amniotic compartment. We propose that lactoferrin is part of the repertoire of host defense mechanisms against intra-amniotic infection. PMID- 11035336 TI - Validation of bioimpedance estimates of cardiac output in preeclampsia. AB - OBJECTIVE: We sought to evaluate the capacity of a new thoracic electric bioimpedance system to estimate cardiac output in patients with preeclampsia. STUDY DESIGN: We performed a prospective comparison of thoracic electric bioimpedance and echocardiographic M-mode volumetric estimates of cardiac output (in liters per minute) in preeclampsia. Subjects with preeclampsia who were chosen by means of strict criteria (either a systolic blood pressure >/=140 mm Hg or a diastolic blood pressure 90 mm Hg, or both, and proteinuria >/=300 mg in 24 hours or >/=+1 on repeat dipstick measurement 6 hours apart) were asked to participate in an institutional review board-approved study. Thoracic electric bioimpedance and echocardiography were performed with the patients in the left lateral recumbent position. Thoracic electric bioimpedance estimates were recorded at bedside; investigators were blinded to 3 simultaneously obtained echocardiographic M-mode estimates of cardiac output. Means were entered as the estimate for each patient. To satisfy the assumption of independent samples, only 1 estimate from each technique was used for each patient. Data were analyzed by Bland-Altman comparison. Hemodynamic and demographic variables are presented as mean +/- SD. RESULTS: Fifteen patients were enrolled. Mean maternal age was 25.9 +/- 4.8 years, and mean gestational age was 34.0 +/- 3.5 weeks. Mean arterial pressure was 112 +/- 14 mm Hg. There was good agreement of thoracic electric bioimpedance-derived and M-mode-derived cardiac output estimates. CONCLUSIONS: In patients with preeclampsia, thoracic electric bioimpedance estimates of cardiac output compare well with echocardiographic M-mode estimates. PMID- 11035337 TI - Evidence for the participation of interstitial collagenase (matrix metalloproteinase 1) in preterm premature rupture of membranes. AB - OBJECTIVE: Rupture of membranes is thought to result from the effects of physical forces in localized areas of the membranes weakened by the degradation of structural collagens. Matrix metalloproteinases are enzymes that degrade extracellular matrix components and have been implicated in membrane rupture. The objective of this study was to determine whether spontaneous rupture of membranes is associated with a change in the amniotic fluid concentration of interstitial collagenase (matrix metalloproteinase 1 [MMP-1]), a major collagenase. STUDY DESIGN: A cross-sectional study was conducted to determine MMP-1 concentrations in amniotic fluid from 353 women in the following categories: (1) term with intact membranes not in labor and in labor, (2) preterm labor who delivered at term, (3) preterm labor who delivered preterm without microbial invasion of the amniotic cavity, (4) preterm labor who delivered preterm with microbial invasion of the amniotic cavity, (5) preterm premature rupture of membranes with and without microbial invasion of the amniotic cavity, (6) term premature rupture of membranes not in labor and in labor, and (7) mid trimester of pregnancy. Microbial invasion of the amniotic cavity was determined by an amniotic fluid culture positive for microorganisms. MMP-1 concentrations in amniotic fluid were determined by means of sensitive and specific immunoassays. RESULTS: (1) MMP-1 was detectable in 81.3% of amniotic fluid samples (287/353), and its concentrations increased with advancing gestational age (r = 0.4; P <.001). (2) Preterm premature rupture of membranes was associated with a significant increase in the median amniotic fluid concentration of MMP-1 (P =.02). (3) Women with term premature rupture of membranes had a significantly lower amniotic fluid MMP-1 concentration than those with intact membranes at term not in labor (P <.001). (4) Microbial invasion of the amniotic cavity in patients in preterm labor with intact membranes and in patients with preterm premature rupture of membranes was also associated with significant increases in the median amniotic fluid MMP-1 concentrations (P <.05 and P <.01, respectively). (5) Patients with preterm premature rupture of membranes and microbial invasion of the amniotic cavity had a significantly higher median amniotic fluid MMP-1 concentration than those with intact membranes and microbial invasion of the amniotic cavity (P =.01). (6) Neither term nor preterm parturition was associated with changes in amniotic fluid MMP-1 concentrations (P =.6 and P =.3, respectively). CONCLUSION: (1) Collagenase 1 (MMP-1) is a physiologic constituent of amniotic fluid. (2) Preterm premature rupture of membranes (in both the presence and absence of infection) was associated with an increase in the amniotic fluid MMP-1 concentrations. (3) Neither term nor preterm parturition was associated with a significant increase in the amniotic fluid concentration of MMP-1. PMID- 11035338 TI - Impact of high-dose corticosteroid therapy for patients with HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome. AB - OBJECTIVE: The purpose of this study was to determine whether corticosteroid administration to patients with antepartum HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome would alter laboratory values diagnostic for the disease. STUDY DESIGN: Cases of 37 women with antepartum HELLP syndrome managed between March 1995 and July 1999 were reviewed. Patients were classified on the basis of exposure to corticosteroids and to the dose used. Group 1 did not receive corticosteroids. Group 2 received a standard corticosteroid dosage regimen for promotion of fetal lung maturation. Group 3 received a high-dose corticosteroid regimen of >24 mg/d (most frequently 10 mg dexamethasone as an intravenous bolus dose every 6 hours for 2 doses followed by 6 mg as an intravenous bolus dose every 6 hours for 2 to 4 doses). Antepartum changes in laboratory values from diagnosis to delivery were evaluated by means of the Kruskal-Wallis test. RESULTS: Eleven patients did not receive corticosteroids, 15 were given a standard dose, and 11 received high-dose therapy. For each laboratory value assessed (platelet count, aspartate aminotransferase activity, and lactate dehydrogenase activity), the corticosteroid groups differed significantly from the no-treatment group (P 2 doses of betamethasone were excluded. All patients received broad-spectrum prophylactic antibiotics. Data were analyzed with the Student t test, the chi(2) test, and the Fisher exact test. Multiple logistic regression analyses incorporated multiple variables considered risk factors for respiratory distress syndrome and intraventricular hemorrhage. P <.05 for all 2-tailed tests was considered significant. RESULTS: A total of 362 patients were included, with 203 in the control group and 159 in the single-course group. Patients in these groups were delivered at 31.0 +/- 3.0 and 30.2 +/- 2.7 (mean +/- SD) weeks' gestation, respectively. The groups were similar with respect to selected demographic characteristics, latency until delivery, mode of delivery, birth weight, and maternal group B streptococcal colonization status. Univariate analysis demonstrated significant decreases in the frequencies of both respiratory distress syndrome (odds ratio, 0.31; 95% confidence interval, 0.2-0.5) and grade III/IV intraventricular hemorrhage (odds ratio, 0.14; 95% confidence interval, 0.1-0.6) in the single-course group. The frequencies of early neonatal sepsis, chorioamnionitis, endometritis, and neonatal death were similar between groups. Multiple logistic regression analyses determined that a single course of betamethasone was independently associated with reductions in the frequencies of both respiratory distress syndrome (odds ratio, 0.16; 95% confidence interval, 0.1-0.4) and grade III/IV intraventricular hemorrhage (odds ratio, 0.18; 95% confidence interval, 0.1-0.4). CONCLUSIONS: A single course of betamethasone administered antenatally to patients with preterm premature rupture of membranes was associated with decreases in the frequencies of both respiratory distress syndrome and advanced grades of intraventricular hemorrhage without any increase in perinatal infectious morbidity. PMID- 11035340 TI - Elevated circulating concentrations of platelet activating factor in preeclampsia. AB - OBJECTIVE: The aim of this study was to determine whether any association exists between preeclampsia and circulating platelet activating factor levels. STUDY DESIGN: We performed a cross-sectional observational study of circulating platelet activating factor concentrations in nonpregnant women, normotensive pregnant women in the third trimester, women with preeclampsia in the third trimester, and normotensive men. Platelet activating factor concentrations were measured with a commercially available platelet activating factor-specific radioimmunoassay (NEN Life Science Products, Inc, Boston, Mass). The primary outcome measure was the difference in mean platelet activating factor concentrations among the 4 study groups. Preeclampsia was determined according to the criteria of The American College of Obstetricians and Gynecologists. Data were analyzed with the Student t test, the chi(2) test, the Fisher exact test, analysis of variance, and the Tukey test for pairwise multiple comparisons, with significance established at P <.05. RESULTS: The mean (+/-SD) circulating concentration of platelet activating factor was significantly higher in the group with preeclampsia (338.1 +/- 26.9 ng/mL) than in either the normotensive pregnant group (217.9 +/- 25.9 ng/mL; P <.05) or the nonpregnant female group (237.9 +/- 20.9 ng/mL; P <.05). The 2 pregnant groups were similar with respect to selected demographic characteristics and gestational age at time of collection. There were no significant differences in the mean platelet activating factor concentrations between the group with preeclampsia and the normotensive male group or between the normotensive pregnant female group and the nonpregnant female group. CONCLUSION: Circulating platelet activating factor concentrations were increased in women with pregnancies complicated by preeclampsia with respect to those in normotensive pregnant women and normotensive nonpregnant women. Platelet activating factor may therefore serve as a marker for the risk of preeclampsia. PMID- 11035341 TI - A simplified index of the plasma sodium threshold for arginine vasopressin secretion-morning fasting, euhydrated sodium levels. AB - OBJECTIVE: Human pregnancy results in a reduction in plasma osmolality and thus a reduction in the osmotic threshold for arginine vasopressin secretion. Although the functional characteristics of the osmoregulatory system controlling arginine vasopressin secretion have been carefully defined, determination of the osmotic threshold requires a complex, labor-intensive protocol of an intravenous hypertonic saline infusion. To aid in studies of osmotic threshold resetting in pregnancy, we sought to develop a simplified method for determination of this value. STUDY DESIGN: Ten healthy nonpregnant women between the ages of 18 and 40 years were studied over 2 days. All patients were hospitalized, and morning euhydration was ensured by oral water hydration (5-10 mL/kg) the evening before the study. On the first study day, patients were fed a standard no-salt-added diet; plasma osmolality and sodium values were checked just before and 1 and 2 hours after meals. On the second study day, after fasting blood samples were obtained, patients received an intravenous infusion (0.06 mL. kg(-1). min(-1) for 120 minutes) of hypertonic (5%) saline to gradually increase the plasma sodium level. Blood samples were obtained every 15 minutes for measurement of plasma electrolytes and arginine vasopressin. Plasma arginine vasopressin concentrations were regressed against plasma osmolality and sodium concentration to calculate the osmotic threshold for arginine vasopressin secretion. RESULTS: Hypertonic saline injection significantly increased plasma sodium (from 139 +/- 1 to 149 +/- 1 mEq/L) and osmolality (from 284 +/- 2 to 304 +/- 2 mOsm/kg H(2)O). Plasma arginine vasopressin significantly increased (from 5 +/- 1 to 30 +/- 10 pg/mL). The mean sodium and osmolality thresholds for arginine vasopressin secretion were calculated as 137 +/- 2 mEq/L and 285 +/- 15 mOsm/kg H(2)O. The mean morning fasting sodium level was nearly identical to the calculated sodium threshold, whereas the morning fasting osmolality value was significantly different. CONCLUSIONS: The morning fasting, euhydrated sodium level can be used as a simplified index for the plasma osmotic threshold for arginine vasopressin secretion. This index may provide a useful predictive measure for pregnant women in whom the plasma volume does not expand. PMID- 11035342 TI - Outcome of pregnancies complicated by ruptured membranes after genetic amniocentesis. AB - OBJECTIVE: We sought to compare perinatal outcomes of pregnancies complicated by preterm premature rupture of membranes after genetic amniocentesis with pregnancies complicated by spontaneous preterm premature rupture of membranes at a similar gestational age. STUDY DESIGN: A retrospective study was performed in which a computerized database was reviewed to identify all patients presenting to our institution with preterm premature rupture of membranes within 48 hours of a genetic amniocentesis from July 1988 to August 1999. Control subjects were matched for gestational age at preterm premature rupture of membranes. Patients were all managed expectantly. Outcomes were compiled from review of medical records. Descriptive statistics, the Student t test, and the chi(2) test were used, with P <.05 considered significant. RESULTS: During the study period, genetic amniocentesis was performed 1101 times. Eleven (1%) women presented within 48 hours with preterm premature rupture of membranes. The mean gestational age at the time of rupture was not different between the cases in which preterm premature rupture of membranes occurred after genetic amniocentesis compared with the control subjects in whom preterm premature rupture of membranes occurred spontaneously (16.5 weeks vs 17.6 weeks, respectively). Women with preterm premature rupture of membranes after amniocentesis experienced significantly longer latency periods (124 vs 28 days; P =.0001) and delivered at more advanced gestational ages (34.2 vs 21.6 weeks; P =.0002) than those with spontaneous preterm premature rupture of membranes. The perinatal survival rate was 91% in pregnancies complicated by preterm premature rupture of membranes after genetic amniocentesis compared with a rate of 9% in control subjects (P =.005). CONCLUSIONS: Pregnancies complicated by preterm premature rupture of membranes after genetic amniocentesis result in significantly better perinatal outcomes compared with pregnancies complicated by spontaneous preterm premature rupture of membranes at a similar gestational age. Expectant management should be considered in such cases. PMID- 11035343 TI - Magnesium sulfate therapy affects attention and working memory in patients undergoing preterm labor. AB - OBJECTIVE: Patients commonly consent to undergoing invasive procedures while receiving magnesium sulfate therapy. This study evaluated the effects of magnesium sulfate on attention, comprehension, and memory in patients undergoing preterm labor. STUDY DESIGN: Consenting patients were studied while receiving(study) and not receiving (control) intravenous magnesium sulfate tocolysis for preterm labor. Excluded were patients with possible preeclampsia, imminent delivery, sedative administration, or prior mental illness. Patient comprehension was assessed with the Boston Diagnostic Aphasia Examination. Level of attention and working memory were evaluated with the Paced Auditory Serial Addition Test. Verbal learning, short-term memory, and recognition were determined with the Hopkins Verbal Learning Test. Gross mental-neurologic deficits were evaluated with the Mini-Mental Status Examination. The tests were administered by the same examiner. Control testing was performed >24 hours after intravenous magnesium sulfate was discontinued. Magnesium levels were obtained at the time of testing. The primary outcome measure was the Paced Auditory Serial Addition Test score because of its ability to elicit subtle differences in attention capacity. Statistical analysis included the paired t test and the McNemar test. RESULTS: Fifteen patients completed the study. Paced Auditory Serial Addition Test scores were significantly higher (ie, more errors were made) during magnesium sulfate therapy than periods without therapy (14 +/- 8 vs 7 +/- 7; P <.05). Comprehension (Boston Diagnostic Aphasia Examination score) was not different between the groups (P =.7). There was no difference in short-term memory (Hopkins Verbal Learning Test) or gross mental-neurologic deficits between the 2 groups (all P >.1). CONCLUSIONS: Magnesium sulfate therapy appears to have an effect on attention and working memory but not on long-term memory or comprehension. The significant differences in Paced Auditory Serial Addition Test scores reveal deficits in information-processing ability in patients on a regimen of magnesium sulfate therapy. PMID- 11035344 TI - Low-dose folic acid supplementation reduces plasma levels of the cardiovascular risk factor homocysteine in postmenopausal women. AB - OBJECTIVE: The aim of our randomized, controlled trial was to verify the effect of folic acid supplementation on homocysteine levels in postmenopausal women. STUDY DESIGN: Thirty-six women were divided randomly into 2 groups as follows: a placebo group (n = 18) and a group receiving 500 microg folic acid per day for 4 weeks (n = 18). To assess concentrations of plasma homocysteine, venous blood samples were taken on enrollment and after 4 weeks of treatment. RESULTS: Mean plasma homocysteine levels were 10.9 +/- 2.7 micromol/L in the placebo group and 7.8 +/- 2.35 micromol/L (P <.01) in the group receiving 500 microg folic acid per day for 4 weeks. The thirds (referred to as tertiles) of women with the highest baseline homocysteine plasma levels showed the greatest reduction in homocysteine, with a mean decrease of 4.35 micromol/L (32%; P <.01), in comparison with a decrease of 3.35 micromol/L (29%; P <.01) in the middle tertile and 1.3 micromol/L (22.4%; P =.09) in the lower tertile. CONCLUSIONS: The results show that low doses of folic acid are associated with a significant reduction in plasma concentrations of homocysteine. The highest initial levels of homocysteine showed the most important reduction after therapy. PMID- 11035345 TI - Cervical shedding of herpes simplex virus and cytomegalovirus throughout the menstrual cycle in women infected with human immunodeficiency virus type 1. AB - OBJECTIVE: Our purpose was to evaluate the frequency and patterns of the shedding of herpes simplex virus and cytomegalovirus in the female genital tract throughout the menstrual cycle. STUDY DESIGN: Seventeen women, all seropositive for herpes simplex virus types 1 and 2, cytomegalovirus, and human immunodeficiency virus type 1, underwent daily evaluation of cervical viral shedding for the duration of 1 menstrual cycle (21-31 visits per woman). Serum estradiol and progesterone levels were monitored 3 times weekly. RESULTS: Overall, herpes simplex virus deoxyribonucleic acid was detected in 43 (10%) of 450 cervical swabs, and cytomegalovirus deoxyribonucleic acid was detected in 232 (52%) of 450 cervical swabs. For individual women there was considerable variability in the percentage of days on which virus was detected, ranging from 0% to 33% for herpes simplex virus and from 20% to 97% for cytomegalovirus. Shedding of herpes simplex virus did not vary significantly with menstrual cycle; however, shedding of cytomegalovirus was significantly more frequent in the luteal phase (odds ratio, 1.9; 95% confidence interval, 1.1-3.4). A CD4(+) lymphocyte count <200/microL was associated with increased frequency of the detection of herpes simplex virus (odds ratio, 5.7; 95% confidence interval, 1.1 29.4). CONCLUSIONS: Asymptomatic cervical shedding of both herpes simplex virus and cytomegalovirus occurs very frequently in women infected with human immunodeficiency virus type 1. The risk of transmitting these viruses to sexual partners and neonates may be higher than previously recognized. PMID- 11035346 TI - Magnetic resonance imaging in an experimental model of human ovarian cancer demonstrating altered microvascular permeability after inhibition of vascular endothelial growth factor. AB - OBJECTIVE: Magnetic resonance imaging enhanced with macromolecular contrast medium was used to monitor effects of angiogenesis inhibition on tumor microvascular permeability and ascites volume in an athymic rat model of human ovarian cancer. STUDY DESIGN: Groups of 6 athymic rats implanted intraperitoneally with SKOV-3, a human ovarian cancer cell line, were treated through a 14-day course with antibody to vascular endothelial growth factor or with saline solution for control animals. Dynamic magnetic resonance imaging was performed with a 92,000-d contrast agent, albumin-(gadolinium diethylenetriaminepentaacetic acid)(30). Vascular permeability was estimated from dynamic enhancement data that were analyzed with a unidirectional 2-compartment kinetic model. RESULTS: Animals treated with vascular endothelial growth factor antibody accumulated significantly smaller volumes of peritoneal ascites (P <.05) and showed significantly lower magnetic resonance imaging-assayed tumor microvascular permeabilities (P <.05) than did control animals. CONCLUSION: Magnetic resonance imaging enhanced with a macromolecular contrast agent in an athymic rat model of human ovarian cancer treated with anti-vascular endothelial growth factor antibody can be used to measure a reduction in tumor microvascular permeability, corresponding to a reduction in ascites production. PMID- 11035347 TI - The fate of rectus fascia suburethral slings. AB - OBJECTIVE: Autologous rectus fascia is commonly used to construct suburethral slings for the treatment of genuine stress incontinence. This fascia performs well and has not been associated with clinical problems related to its choice as a sling material. However, the histologic appearance of such slings after implantation has not been documented. STUDY DESIGN: At the time of revision of autologous rectus fascia suburethral slings in 5 patients, biopsy specimens of the slings were obtained and submitted for histologic examination. A specimen of rectus fascia before implantation was also obtained from a sixth patient who had no symptoms. RESULTS: After implantation autologous rectus fascia slings remain viable. There is fibroblast proliferation, neovascularization, and remodeling of the graft. No evidence of inflammatory reaction or of graft degeneration was detected. A linear orientation of connective tissue and fibroblasts was seen in some areas, whereas other areas had remodeled to form tissue similar to noninflammatory scar. CONCLUSION: Autologous rectus fascia slings undergo extensive remodeling after implantation. PMID- 11035348 TI - Epithelial cell layer thickness and immune cell populations in the normal human vagina at different stages of the menstrual cycle. AB - OBJECTIVE: The aim of our study was to examine vaginal tissue during 3 phases of the menstrual cycle for the number of cell layers and epithelial immune cells. STUDY DESIGN: Vaginal biopsies were performed during 3 phases of the normal menstrual cycle (menstrual, days 1-5; preovulatory, days 7-12; and postovulatory, days 19-24) in 74 subjects. A subset of women had vaginal tissues stained with specific monoclonal antibody markers for Langerhans cells (CD1a), macrophages (KP1), T and B lymphocytes (CD4, CD8, CD21) and neutrophils (CD15). The number of cell layers and the number of immune cells in the vaginal tissue biopsy specimen were determined by a single observer who was blinded to clinical data. RESULTS: At 3 phases of the normal menstrual cycle, the mean number of epithelial cell layers underwent a small but statistically significant decrease from 27.8 +/- 0.7 on days 1-5 and 28.1 +/- 0.6 on days 7-12 to 26.0 +/- 0.7 on days 19-24 of the cycle (P =.01). Nonovulating women had a reduced mean epithelial cell layer count on days 7-12 (23.7 +/- 1. 4) compared with the epithelial cell layer count in ovulating women (28.8 +/- 0.7; P =.005). No significant changes were observed in the mean number per high-power field of Langerhans cells, macrophages, CD4 or CD8 lymphocytes, and neutrophil cell populations during the 3 phases of the cycle. B lymphocytes were not observed in the vaginal tissues. CONCLUSION: A small but statistically significant reduction in the number of vaginal epithelial cells was observed over the menstrual cycle. This reduction is not likely to be clinically significant. Immune cell populations in the vaginal tissues appeared stable throughout the menstrual cycle. PMID- 11035349 TI - The effect of a rapid change in availability of epidural analgesia on the cesarean delivery rate: a meta-analysis. AB - OBJECTIVE: This study was undertaken to quantitatively estimate the effect of a rapid introduction or withdrawal of on-demand epidural analgesia on the cesarean delivery rate. STUDY DESIGN: MEDLINE and meeting abstracts were searched for studies reporting the cesarean delivery rate immediately before and after a rapid change in the availability of epidural analgesia. Nine studies reporting data on 37,753 patients were selected. Meta-analysis was performed to estimate the means and 95% confidence intervals for the changes in rates of total cesarean deliveries, cesarean deliveries among nulliparous women, cesarean deliveries for dystocia, and operative vaginal deliveries. RESULTS: There was no significant change in the overall cesarean delivery rate with an increase in the availability of epidural analgesia. Similarly, the rates of cesarean deliveries among nulliparous patients, of cesarean deliveries for dystocia, and of operative vaginal deliveries did not significantly differ between periods of high and low epidural analgesia availability. CONCLUSION: A rapid change in the availability of epidural analgesia is not associated with any increase in the cesarean delivery rate. PMID- 11035350 TI - Pretreatment of human amnion-chorion with vitamins C and E prevents hypochlorous acid-induced damage. AB - OBJECTIVE: Preterm premature rupture of fetal membranes has been associated with infection, cigarette smoking, and bleeding. Hypochlorous acid (a reactive oxygen species) is central to the body's response to infection, yet it may damage surrounding tissue while destroying pathogens. We examined in vitro the tissue damaging actions of hypochlorous acid on the amnion-chorion and the protective role provided by pretreatment with vitamins C and E. STUDY DESIGN: Amnion-chorion samples were obtained from 4 term pregnancies, cut into segments, and divided into 6 exposure groups. Half were treated in advance with vitamins C and E (Trolox C) and half were treated with buffer solution alone. After rinsing, amnion-chorion samples were exposed to hypochlorous acid at 1 or 10 mmol/L for 4 hours. Histologic and immunocytochemical evaluations were conducted with antibodies for collagen I and IV. RESULTS: Extensive damage to amniotic epithelium and collagen I but not collagen IV resulted from hypochlorous acid exposure and was dose related. Pretreatment with vitamins C and E prevented this damage in all cases. CONCLUSION: Hypochlorous acid damages the amniotic epithelium and collagen I in the amnion-chorion. The protection against hypochlorous acid-induced damage provided by antioxidant therapy (vitamins C and E) is of therapeutic significance. PMID- 11035351 TI - Maternal and neonatal outcomes after induction of labor without an identified indication. AB - OBJECTIVE: This study was undertaken to examine associations between induction of labor and maternal and neonatal outcomes among women without an identified indication for induction. STUDY DESIGN: This was a population-based cohort study of 2886 women with induced labor and 9648 women with spontaneous labor who were delivered at 37 to 41 weeks' gestation, all without identified medical and obstetric indications for induction. RESULTS: Among nulliparous women 19% of women with induced labor versus 10% of those with spontaneous labor underwent cesarean delivery (adjusted relative risk, 1.77; 95% confidence interval, 1.50 2.08). No association was seen in multiparous women (relative risk, 1.07; 95% confidence interval, 0. 81-1.39). Among all women induction was associated with modest increases in instrumental delivery (19% vs 15%; relative risk, 1.20; 95% confidence interval, 1.09-1.32) and shoulder dystocia (3.0% vs 1. 7%; relative risk, 1.32; 95% confidence interval, 1.02-1.69). CONCLUSION: Among women who lacked an identified indication for induction of labor, induction was associated with increased likelihood of cesarean delivery for nulliparous but not multiparous women and with modest increases in the risk of instrumental delivery and shoulder dystocia for all women. PMID- 11035352 TI - The effect of fetal neck position on nuchal fold thickness. AB - OBJECTIVE: Our purpose was to determine whether ultrasonographic measurements of nuchal fold thickness are affected by the position of the fetal neck. STUDY DESIGN: Fetal nuchal fold thickness was prospectively measured in 258 women undergoing routine ultrasonography at 15 to 21 completed weeks of gestation. Patients with fetal structural or chromosomal anomalies were excluded. At the time of examination the position of the fetal head was noted as being extended or flexed on the basis of the angle between the spine and the base of the skull. Gestational age was based on menstrual dates or ultrasonographic biometric parameters. Data were tested for normality. Mann-Whitney U test and analysis of covariance were used (significance was considered to be P <.05). Data are presented as median and range. RESULTS: A total of 258 fetuses were examined with 167 (65%) in the flexed and 91 (35%) in the extended neck position. Gestational age was not significantly different between the flexed and extended groups (median, 19.1 weeks; range, 15.5-21.6 weeks; vs median, 19.1 weeks; range, 15.6 22 weeks; P =.23). Nuchal fold thickness was significantly lower in the flexed group than in the extended group (median, 3.5 mm; range, 1.3-6.2 mm; vs median, 3.9 mm; range, 2.2-4.9 mm; P =.0097). Nuchal fold thickness increased significantly with gestational age in both groups. The difference in nuchal fold thickness between the 2 groups persisted even after the increase in nuchal fold thickness was adjusted for with gestational age (P =.002, analysis of covariance). The difference between the 2 groups was higher at earlier gestations. CONCLUSION: Nuchal fold thickness is affected by gestational age and fetal neck position. Correction for these variables may improve the accuracy of nuchal fold thickness measurements in screening for fetal chromosomal anomalies. PMID- 11035353 TI - Mifepristone versus vaginally administered misoprostol for cervical priming before first-trimester termination of pregnancy: a randomized, controlled study. AB - OBJECTIVE: This study was undertaken to compare the effectiveness of mifepristone orally administered at 24 or 48 hours before first-trimester vacuum aspiration abortion with that of vaginally administered misoprostol as a cervical priming agent. STUDY DESIGN: In a randomized comparative trial 90 women who requested surgical termination of pregnancy were randomly assigned to receive 200 mg mifepristone orally 24 or 48 hours before the operation or 800 microg misoprostol vaginally 2 to 4 hours before the operation. The main outcome measures were baseline cervical dilatation, cumulative force required to dilate the cervix to 9 mm, and intraoperative blood loss. RESULTS: The baseline cervical dilatation was significantly greater among women who received mifepristone 48 hours before the operation (P =.02). This group also required the least mechanical force to dilate the cervix (P =.06). There were no significant differences among the 3 groups in the intraoperative blood loss, in the operating time, or in patient acceptability. Side effects such as hot flushes and headaches were significantly higher among women who received mifepristone 24 or 48 hours before the operation than among those who received misoprostol (P =.01 and P =. 002, respectively). CONCLUSION: Mifepristone is an effective cervical priming agent when orally administered 48 hours before vacuum aspiration for termination of first-trimester pregnancy. Because of its cost and availability in comparison with misoprostol, however, selective use may have to be considered. PMID- 11035354 TI - The impact of digital cervical examination on expectantly managed preterm rupture of membranes. AB - OBJECTIVE: The purpose of this study was to examine the effects of digital cervical examination on maternal and neonatal outcomes among women with preterm rupture of membranes. STUDY DESIGN: This analysis includes data from a previously reported trial of antibiotic treatment during expectant management of rupture of membranes at 24 to 32 weeks' gestation in singleton and twin gestations. Patients from both the randomized trial (n = 299 in the antibiotic group and n = 312 in the placebo group) and the observational component (n = 183) are included in this analysis. The groups were divided into those with one (n = 161) or two digital cervical examinations (n = 27) and those with no digital cervical examinations (n = 606). RESULTS: The gestational ages at enrollment were similar in the two groups (29 +/- 2 weeks' gestation for one or two examinations vs 29 +/- 2 weeks' gestation for no examinations; P =.85). There were no differences in chorioamnionitis (27% vs 29%; P =.69), endometritis (13% vs 11%; P =.5), or wound infection (0.5% vs 1%; P >.999) between the group with one or two examinations and the no-examination group. Infant outcomes were also similar in the two groups, including early sepsis (6% vs 5%; P =.68), respiratory distress syndrome (51% vs 45%; P =.18), intraventricular hemorrhage (7% vs 7%; P =.67), necrotizing enterocolitis (5% vs 3%; P =.19), and perinatal death (7% vs 5%; P =.45). A composite outcome made up of these neonatal outcomes was not different (56% vs 48%; P =.10) between the group with one or two examinations and the no examination group. The time from rupture to delivery was shorter in the digital examination group (median value, 3 vs 5 days; P <. 009). Multivariable analysis to adjust for antibiotic study group, group B streptococcal culture status, race, and maternal transfer did not modify these results. CONCLUSION: Performance of one or two digital cervical examinations during the course of expectant management of rupture of membranes between 24 and 32 weeks' gestation was associated with shorter latency but did not appear to worsen either maternal or neonatal outcome. PMID- 11035355 TI - Does aspirin have a role in improving pregnancy outcome for women with the antiphospholipid syndrome? A randomized controlled trial. AB - OBJECTIVE: This pilot investigation was undertaken to assess the efficacy of low dose aspirin therapy for the treatment of women with antiphospholipid antibodies when recurrent miscarriage is the only sequela. STUDY DESIGN: A double-blind, randomized, placebo-controlled trial was conducted in the setting of the recurrent miscarriage clinic of a tertiary referral obstetric hospital. The participants were 50 women with a history of recurrent miscarriages (>/=3) and antiphospholipid antibodies. Women with systemic lupus erythematosus or a history of thrombosis were excluded. Women were recruited after full investigative screening at the recurrent miscarriage clinic. Women with >/=3 fetal losses and persistently positive results for antiphospholipid antibodies were randomly allocated to receive either aspirin (75 mg daily) or placebo. Investigators, clinicians, and patients were blinded to the treatment. Rates of live births, antenatal complications, and delivery and neonatal outcomes were recorded prospectively. Data were compared by chi(2) analysis with Yates' correction, the Fisher exact test, or the Student t test as appropriate. RESULTS: There were 10 exclusions after random assignment because of inappropriate inclusion. Eighty five percent of the placebo (17/20) group and 80% of the aspirin-treated group (16/20) were delivered of live infants. This difference was not significant. There were no significant differences in antenatal complications or neonatal morbidity between the groups. CONCLUSIONS: This preliminary study suggests that low-dose aspirin has no additional benefit when added to supportive care for women for whom recurrent early fetal loss is the only sequela of the antiphospholipid syndrome. This live birth rate with supportive care alone exceeds the published live birth rates for women with antiphospholipid antibody mediated recurrent fetal loss who were treated with heparin or corticosteroids. This trial, like all other trials in this field, is small, but its results bring into question the need for pharmacologic intervention for women with antiphospholipid syndrome for whom recurrent fetal loss is the only sequela. Our results highlight the need for a large randomized controlled trial to identify the optimal treatment for this group of women and justify the inclusion of a placebo arm in any such trial. PMID- 11035356 TI - Relation between damage to the placenta and the fetal brain after late-gestation placental embolization and fetal growth restriction in sheep. AB - OBJECTIVE: Our aim was to determine the effects of 30 days of placental insufficiency on fetal brain development and to relate placental damage to the degree of fetal brain injury. STUDY DESIGN: Umbilicoplacental embolization was induced from 110 to 140 days of gestation (term, 147 days) in 7 fetal sheep, such that fetal arterial oxygen saturation was maintained at 50% of pre umbilicoplacental embolization values. Six control fetuses were used. At 140 days the fetal brains and placentas were subjected to structural and histochemical analysis. RESULTS: During umbilicoplacental embolization, fetal arterial oxygen saturation, PaO(2), and pH were reduced (P <.05). Thirty days of umbilicoplacental embolization caused a decrease in cross-sectional area of the placentome (P <.05), with 20% of tissue showing damage. All umbilicoplacental embolization fetuses were growth restricted and had brain damage, most prominently in the cerebral white matter. There was no relation between the extent of placental damage and the severity of fetal brain damage. CONCLUSIONS: The absence of a correlation between damage to the placenta and fetal brain is likely to be caused by variations between individuals in (1) the amount of placenta that is required to be functionally damaged to achieve the prescribed level of hypoxemia and (2) the response of the fetal brain to that level of hypoxemia. PMID- 11035357 TI - Antenatal factors at diagnosis that predict outcome in twin-twin transfusion syndrome. AB - OBJECTIVE: We sought to identify clinical factors at diagnosis that predict outcome in twin-twin transfusion syndrome. STUDY DESIGN: In this retrospective series 23 patients with twin-twin transfusion syndrome were seen in a tertiary referral fetal medicine center over a 3-year period. Ten antenatal factors were assessed to determine their ability to predict outcome by use of ordered logistic regression. These factors were the following: (1) absent or reversed end diastolic flow in the umbilical artery, nonvisible bladder, anhydramnios, and estimated fetal weight of <3rd percentile in the donor; (2) pulsatile umbilical vein, either absent or reversed end-diastolic flow in the ductus venosus, or both, and tricuspid-mitral valve regurgitation in the recipient; and (3) gestational age at presentation, estimated fetal weight discordancy, absent arterioarterial anastomosis, and spontaneous rupture of the membranes or cervical change as pregnancy factors. Management comprised serial amnioreduction (n = 10), selective feticide (n = 5; 4 also had amnioreduction), septostomy (n = 4; 1 also had amnioreduction), and delivery (n = 2). Two patients miscarried before treatment. RESULTS: The chance of survival of both twins fell and double deaths increased linearly with increasing number of adverse factors (P =.026). A low chance of survival was independently associated with absent or reversed end diastolic flow in the donor umbilical artery (P =.02) and with a pulsatile umbilical vein or absent or reversed end-diastolic flow in the ductus venosus (P =.03) of the recipient. The probability of at least one twin surviving was only 33% if there was absent or reversed end-diastolic flow in the donor umbilical artery or 37% when abnormal venous recordings were seen in the recipient. An arterioarterial anastomosis detected at diagnosis also influenced prognosis, with all twins surviving when an arterioarterial anastomosis was identified (P =.04). CONCLUSIONS: Three factors identified at diagnosis independently predict poor survival in twin-twin transfusion syndrome-absent or reversed end-diastolic flow in the donor umbilical artery, abnormal pulsatility of the venous system in the recipient, and absence of an arterioarterial anastomosis. These may have a role in the counseling of parents and in selecting the appropriate treatment strategy. PMID- 11035358 TI - Normal development of human fetal hematopoiesis between eight and seventeen weeks' gestation. AB - OBJECTIVE: The aim of this study was to compare the hematologic compositions of fetal blood and liver and to phenotypically quantify the hematopoietic stem and progenitor cells during early human gestation. STUDY DESIGN: Fifty fetal blood samples and 50 fetal livers were collected at 10 to 17 weeks' gestation and 8 to 17 weeks' gestation, respectively. Investigations included fetal blood cell counts, determinations of red blood cell index values, and flow cytometric analyses of mononuclear cells. RESULTS: Fetal red blood cell, white blood cell, and platelet counts all increased with gestation, reflecting hematologic development. The proportion of normoblasts decreased dramatically with gestation. Individual mature red blood cells were larger and contained more hemoglobin during early gestation. Circulating and hepatic T lymphocytes increased in number shortly before the 13th week of gestation, which reflected thymic maturation. As a proportion fetal liver contained fewer T lymphocytes than did fetal blood (2.5% vs 18.6%; P =.003) but more CD34(+) hematopoietic stem and progenitor cells (17.5% vs 4.3%; P =. 004). As a proportion, fetal liver contained more of the primitive CD34(+) and CD38(-) hematopoietic stem and progenitor cells than did fetal blood (32% vs 17%; P =.04). CONCLUSION: Both fetal blood and liver provide a rich source of hematopoietic stem and progenitor cells. Fetal liver provides a richer source of more primitive hematopoietic stem and progenitor cells than does fetal blood. For stem cell transplantation we suggest that fetal livers be collected before the 13th week of gestation, because T lymphocytes are present in much greater numbers in the fetal liver after this stage of gestation. Further, we suggest that in utero stem cell transplantations in fetuses with normal immune development should be performed before the 13th week of gestation. PMID- 11035359 TI - Increase of the isoprostane 8-isoprostaglandin f2alpha in maternal and fetal blood of rats with streptozotocin-induced diabetes: evidence of lipid peroxidation. AB - OBJECTIVE: Pregnancy complicated by diabetes is associated with maternal complications and fetal abnormalities. Animal models of diabetes suggest that heightened free radical production may be implicated in the pathogenesis of this condition. The purpose of this investigation was to evaluate oxidative stress in plasma from diabetic rats and their fetuses through measurement of concentrations of 8-isoprostaglandin F(2alpha), a stable marker of lipid peroxidation. STUDY DESIGN: Diabetes was induced in virgin and pregnant rats with streptozotocin. Blood samples were collected after 20 days of diabetes. Adult and fetal plasma 8 isoprostaglandin F(2alpha) concentrations were determined by gas chromatography mass spectroscopy. RESULTS: Significantly higher plasma 8-isoprostaglandin F(2alpha) concentrations were observed in the virgin rats with diabetes and in both the pregnant dams with diabetes and their fetuses when compared with their respective control groups without diabetes (P <.001). CONCLUSION: Oxidative stress was induced in both mother and fetus in rodent pregnancy complicated by diabetes. This finding may have implications for fetal dysmorphogenesis and in fetal programming for adulthood disease. PMID- 11035360 TI - A faculty development course in obstetrics and gynecology. AB - Sponsored by the nonprofit Berlex Foundation, this 6-day Faculty Development Course introduces small groups of new faculty to research design, critical appraisal of the literature, scientific writing, and evidence-based medicine. The course includes didactic presentations, small group discussions, and individual protocol development. Its influence on academic careers is impossible to determine, because defining an appropriate comparison group is not feasible. However, the cumulative effect should be to increase the research expertise of some young faculty in academic departments nationwide. In addition, many participants have introduced the principles of evidence-based medicine in their own teaching programs. PMID- 11035361 TI - Transperineal versus transvaginal ultrasonography during the second trimester. PMID- 11035363 TI - Cigarette smoking during pregnancy. PMID- 11035365 TI - Were there any misdiagnoses? PMID- 11035367 TI - Cytopathology or immunopathology? The puzzle of cytomegalovirus pneumonitis revisited. AB - Various hypotheses have been proposed to explain why cytomegalovirus pneumonitis (CMV-P) is frequent and severe in bone marrow transplant patients while remaining rare and mild in HIV infected patients. One hypothesis suggests that CMV-P is an immunopathological condition that is common in bone marrow transplantation (BMT) under the effects of an abnormally regenerating immune system that reacts against CMV infected lung tissue. Such a hypothesis implicates CD4 T lymphocytes as one of the critical cell populations involved in immunopathology and also suggests that this process would be aborted by CD4 T cell deficiency in HIV infection. However, studies correlating the onset of CMV-P with lymphocyte reconstitution following BMT have revealed that CD4 cells are present at very low frequencies in the blood during the early period after transplantation when most cases of CMV-P occur. Furthermore, studies directly investigating bronchoalveolar lavage cell types during episodes of CMV-P in BMT patients have also failed to demonstrate significant CD4 involvement and, instead, have emphasized a predominance of natural killer (NK) cells and CD8 cells. These findings serve as the basis for questioning the validity of a CD4-driven immunopathological model of CMV-P in BMT. On the other hand, a variety of experimental and clinical observations support the protective role of CMV-specific CD3+ CD8 T lymphocytes against CMV in both immunocompetent individuals and BMT patients. In a murine BMT model, adoptive transfer of syngeneic BM cells was associated with massive increases in lung CD8 cells which resulted in the resolution rather than the exacerbation of existing CMV-P. In the light of these findings a more plausible hypothesis for CMV-P in BMT is that during the early period after transplantation adequate protective CD8 responses are absent and an uncontrolled CMV proliferation is allowed to develop. Once a critical viral load is reached a cytokine 'storm' may be triggered in the lung tissue that aggravates direct CMV-associated cytopathic effects. Likely candidates for this process would include the release of tumour necrosis factor-alpha (TNF-alpha) from alveolar macrophages stimulated by interferon-gamma (IFN-gamma) released from NK cells that are reconstituted early after BMT. PMID- 11035368 TI - High-dose thiotepa and hematopoietic stem cell transplantation in pediatric malignant mesenchymal tumors: a phase II study. AB - The prognosis of metastatic malignant mesenchymal tumors (MMT) remains poor. Given the chemosensitivity of these neoplasms, a phase II study of high-dose thiotepa (HDT) was performed to evaluate the efficacy of this drug in this particular subset of pediatric tumors. Between 1986 and 1998, 18 patients, previously treated with conventional therapy for metastatic or refractory MMT, entered the study. Thiotepa was administered at a daily dose of 300 mg/m2 for 3 consecutive days. Hematopoietic stem cell rescue, consisting of bone marrow transplantation or peripheral stem cell transplantation, was performed 2 days after completion of HDT. A response exceeding 50% was observed in 6/18 patients (response rate 33%). Toxicity was severe but never led to death. HDT used at a dose of 900 mg/m2 yields measurable anti-tumor activity in previously treated patients. The next step in these particularly poor prognosis metastatic MMT will be to investigate HDT combined with other drugs, known to be efficient at high doses. PMID- 11035369 TI - Maintenance of immune memory to the hepatitis B envelope protein following adoptive transfer of immunity in bone marrow transplant recipients. AB - Adoptive transfer of immunity against hepatitis B surface antigen (HBsAg) has been documented in mice and humans. In the present study, we report long-term follow-up of antibodies to HBsAg in humans who received allogeneic bone marrow transplantation (BMT) from donors immunized with HBsAg. BM donors were immunized with recombinant HBsAg. BM or PB cells were transplanted to HLA matched recipients. Recipients were followed for anti-HBs seroconversion. Control groups included non-immunized or rHBsAg immunized healthy adults as well as individuals that had had hepatitis B and recovered spontaneously. PBLs were stimulated in vitro with rHBsAg and stimulation was expressed as stimulation index. Adoptive transfer of immunity to HBsAg was initially documented in 12 recipients of BM from anti-HBc+/anti-HBs+ donors. An almost 4 year follow-up showed detectable protective anti-HBs levels (>10 mIU/ml) in 50% of patients. Immunity to HBV was also documented in 22/35 BMT recipients (62%), who received their bone marrow from actively immunized donors. In 7/9 of these BMT recipients, anti-HBs antibodies levels were documented 25 months following BMT. In 6/8 (75%) of patients who received only PBLs from HBV immune donors, adoptive transfer of immunity to HBV, and seroconversion to HBsAg+, were documented within 2 months of i.v. injection. Evidence for specific cellular immune response with increased SIs was documented for healthy vaccinees, and BMT recipients, and in none of the healthy non-vaccinated controls. These results suggest that adoptive transfer of immunity to HBV is a useful method for providing long-lasting protection for BM recipients. PMID- 11035371 TI - The spectrum of Malassezia infections in the bone marrow transplant population. AB - A consecutive series of 3044 patients who underwent BMT at the University of Minnesota over a 25 year period were reviewed for the post-transplant occurrence of infection caused by the yeast Malassezia furfur. Six patients, ranging in age from 1 to 54 years, developed Malassezia infections at a median of 59 days post transplant. Five patients were allogeneic transplant recipients; the remaining patient had undergone autologous transplantation. A spectrum of clinical manifestations of Malassezia infection was seen in these patients, including infections of mucosal surfaces and the skin, in addition to catheter-related fungemia. Unlike many of the other more common opportunistic fungal infections in immunocompromised patients, neutropenia and the use of broad-spectrum antimicrobials do not appear to be significant risk factors for Malassezia infections in the BMT population. In addition, disseminated fungal infection despite the presence of fungemia is uncommon. Lastly, the outcome of Malassezia infections in these patients, whether folliculitis, mucosal infection, or fungemia, appears to be quite favorable, in contrast to the poorer outcome with many other fungal infections in BMT patients. Catheter removal and discontinuation of intravenous lipids are important for a successful outcome in fungemic cases. PMID- 11035370 TI - Human herpesvirus 6 is an important pathogen in infectious lung disease after allogeneic bone marrow transplantation. AB - Two hundred and ten bronchoalveolar lavage (BAL) samples were obtained from 50 patients 10 days before and on defined days after allogeneic bone marrow transplantation (BMT). The samples were examined for human cytomegalovirus (HCMV) and human herpesvirus-6 (HHV-6) by polymerase chain reaction (PCR). Fifteen patients (30%) had a positive result for HCMV in at least one sample and 25 (50%) were positive for HHV-6 in at least one sample. Five patients developed HCMV associated interstitial pneumonia (HCMV-IP) within 100 days after allogeneic BMT. Four of these patients were positive for both HCMV and HHV-6. Conspicuous HHV-6 positivity was detected in BAL samples obtained because of respiratory symptoms. No association was found between detection of HHV-6 and acute graft-versus-host disease. Engraftment failure or a delay in engraftment was observed in none of the 50 patients. The data from this study indicate that HHV-6 is a pathogen in HCMV-associated, as well as in non-HCMV-associated infectious lung disease after BMT. PMID- 11035372 TI - Long-term liver dysfunction after allogeneic bone marrow transplantation: clinical features and course in 61 patients. AB - This retrospective study has aimed at determining the prevalence, aetiology and clinical evolution of chronic liver disease (CLD) after allogeneic bone marrow transplantation (BMT). A total of 106 patients who had been transplanted in a single institution and who had survived for at least 2 years after BMT were studied. The prevalence of CLD was 57.5% (61/106). In 47.3% of cases more than one aetiopathogenic agent coexisted. The causes of CLD were iron overload (52.4%), chronic hepatitis C (47.5%), chronic graft-versus-host disease (C-GVHD) (37.7%), hepatitis B (6.5%), non-alcoholic steatohepatitis (NASH) (4.9%), autoimmune hepatitis (AIH) (4.9%) and unknown two (3.3%). Twenty-three patients with iron overload underwent venesections which were well tolerated. An improvement in liver function tests (LFTs) was observed in 21 (91%) patients. All six patients with siderosis as the only cause of CLD normalized LFT as well as three patients with HCV infection. Clinical evolution was satisfactory for patients with GVHD, AIH, NASH and hepatitis B. At the last visit 23 patients continued with abnormal LFTs, and 19 of them were infected by the HCV. A sustained biochemical and virologic response was achieved in only one case out of six patients with CHC who received interferon. We have found that CLD is a common complication in long-term BMT survivors. The aetiology is often multifactorial, iron overload, CHC and C-GVHD being the main causes. The CLD followed a rather 'benign' and slow course in our patients as none of them developed symptoms or signs of liver failure and we did not observe an increase in morbidity or mortality in these patients, but a longer follow-up is necessary in HCV infected patients based on the natural history of this infection in other populations. PMID- 11035373 TI - Preparation and analysis of fetal liver extracts. AB - The aim of this work is to describe the techniques that have been used for preparation and analysis of whole fetal liver extracts destined for in utero transplantation. Nine fetal livers between 12 and 17 weeks of gestation were prepared: cell counts and assessment of the hematopoietic cell viability were performed on cell suspensions. Hepatocytes represented 40 to 80% of the whole cell population. The remaining cells were constituted by hematopoietic cells (mainly erythroblasts), as well as by endothelial cells. The latter expressed CD34 on their surface, interfering with the assessment of CD34+ hematopoietic cells by flow cytometry. Direct visual morphologic control using alkaline phosphatase anti-alkaline phosphatase techniques was needed to differentiate hematopoietic from extra-hematopoietic CD34+ cells. Between 3.0 and 34.6 x 10(6) CD34+ viable hematopoietic cells were collected per fetal liver. Adequate differentiation of these cells into burst-forming units erythroid (BFU-E), colony forming units granulocyte-macrophage (CFU-GM), and colony-forming units granulocyte erythroid macrophage megakaryocyte (CFU-GEMM) has been shown for each sample in clonogeneic cultures. In conclusion, fetal liver is a potential source of hematopoietic stem cells. Their numeration, based on the presence of CD34, is hampered by the expression of this antigen on other cells contained in the liver cell extract, in particular endothelial cells. PMID- 11035374 TI - First manifestations of seronegative spondylarthropathy following autologous stem cell transplantation in HLA-B27-positive patients. AB - Two male patients with non-Hodgkin's lymphoma (NHL, follicular NHL, diffuse large B cell NHL, both in 2nd complete remission) and one female patient with acute myeloid leukemia in 1st complete remission developed arthralgias and enthesopathy following autologous stem cell transplantation. In 2/3 patients, sacroiliitis could be demonstrated on X-ray. In both patients, the rheumatic symptoms were classified as manifestations of a spondylarthropathy. All three patients were subsequently shown to be HLA-B27-positive. The patients were successfully treated with non-steroidal anti-inflammatory drugs. The differential diagnosis of joint pain following autologous stem cell transplantation should include HLA-B27 associated spondylarthropathies in addition to the more commonly seen bone and joint pain due to immobilization and medication. PMID- 11035375 TI - Patients with mantle-cell lymphoma relapsing after autologous stem cell transplantation may be rescued by allogeneic transplantation. AB - Two patients with disseminated mantle-cell lymphoma relapsed 24 and 13 months, respectively, after high-dose chemotherapy and autologous stem cell transplantation (autoSCT). Both patients had an HLA-identical sibling and received an allogeneic stem cell transplant (alloSCT) 32 and 18 months after autologous transplant, after conditioning with fractionated 12 Gy total body irradiation plus cyclophosphamide 120 mg/kg. They are both alive and in complete remission 24 months after transplant. Both patients have developed chronic graft versus-host disease and their Karnofsky performance status is 90%. AlloSCT may offer a useful approach in a subgroup of patients with mantle-cell lymphoma who have relapsed after autologous transplantation. PMID- 11035376 TI - Mixed chimera converted into full donor chimera with powerful graft-versus leukemia effects but no graft-versus-host disease after non T cell-depleted HLA mismatched peripheral blood stem cell transplantation. AB - Instead of donor T cell depletion, we used CTLA4 and TJU103 (a small organic compound believed to block CD4 binding to MHC II molecule of APC) to block donor T lymphocyte activation in vitro before infusion, and mycophenolate mofetil to control the activity of lymphocytes of the recipient. We successfully treated a patient with an HLA-mismatched graft without donor T cell depletion. Mixed chimerism was observed 30 days and 60 days after transplantation. STR-PCR showed that 28% and 62% of blood mononuclear cells (MNC) were donor derived at day +30 and day +60, respectively. Mixed chimerism converted into full donor chimerism, when 99.7% of the MNC in the recipient were donor derived after three courses of DLI. A powerful GVL effect related to mixed chimerism was observed. No acute GVHD occurred, only grade II chronic GVHD occurred 6 months after transplant. Based on this case, we suggest that: (1) stable mixed chimerism can be intentionally established across HLA barriers without donor T cell depletion; (2) mixed chimerism can be converted into full donor chimerism by DLI; (3) mixed chimerism induced with this approach can be associated with a very powerful GVL effect, and these may be enhanced by DLI, without severe GVHD. PMID- 11035378 TI - Proceedings of the 1st International Conference on the Management of Erectile Dysfunction. March 3-4, 2000. Chicago, Illinois, USA. PMID- 11035377 TI - Complete remission of idiopathic myelofibrosis following donor lymphocyte infusion after failure of allogeneic transplantation: demonstration of a graft versus-myelofibrosis effect. AB - A patient with idiopathic myelofibrosis (IM) in the osteosclerotic phase received an allogeneic stem cell transplant. Hemopoietic engraftment was rapid, and full donor chimerism was observed on day +70. However, a few months later, replacement of donor hemopoiesis by the patient's 20q- cell clone was observed, followed by reappearance of the blood IM features, marrow fibrosis and osteosclerosis. At 8 months from transplant donor lymphocytes were infused, which induced chronic GVHD. This resulted in normalization of the blood, with disappearance of the fibrosis and osteosclerosis, effects which persisted 20 months later. This case provides evidence for a graft-versus-disease effect in IM. PMID- 11035379 TI - First international conference on the management of erectile dysfunction. Overview consensus statement. PMID- 11035380 TI - The worldwide prevalence and epidemiology of erectile dysfunction. AB - This paper: (i) describes the worldwide prevalence of erectile dysfunction (ED); (ii) presents age-specific incidence rates for ED in the US; (iii) summarizes some key epidemiologic correlates of ED in the general population; and (iv) considers the possibility that ED may be a biobehavioral marker (sentinel event) of subsequent cardiovascular disease in men. Clinical, anthropometric, life style and hormonal data are presented from the milestone Massachusetts Male Aging Study (MMAS), a large (over 1000) prospective cohort of randomly sampled community dwelling, normally aging men. Newly updated population prevalence and (more importantly) age-specific incidence rates are reported. We also estimate the likely magnitude of ED that will accompany the worldwide globalization of aging. Key correlates (predictors) of incident ED, especially vasculogenic influences, are identified and discussed. In conclusion, ED is a common biobehavioral phenomenon and there are strong physiological and epidemiological reasons for considering it a major marker (or predictor) of subsequent cardiovascular disease in men. International Journal of Impotence Research (2000) 12, Suppl 4, S6-S11. PMID- 11035381 TI - Diagnostic evaluation of erectile dysfunction in the era of oral therapy. AB - Therapy for erectile dysfunction (ED) may be specific to the cause of ED or it may be nonspecific. There are only three causes of ED which have specific therapy: psychogenic, endocrine and certain types of reversible vasculogenic ED. In the era of oral therapy for ED, treatment is not cause-specific in the great majority of patients. For this great majority, only the basic evaluation of ED is needed. Only when there is a strong suspicion that the cause of a patient's ED is endocrine, psychogenic or reversible vascular disease are additional diagnostic tests indicated. In these three categories of patients, specific treatment of the cause of ED can produce a permanent and dramatic improvement in sexual function and satisfaction. International Journal of Impotence Research (2000) 12, Suppl 4, S12-S14. PMID- 11035382 TI - Gap junctions and ion channels: relevance to erectile dysfunction. AB - The corporal myocyte is a critical determinant of erectile capacity whose functional integrity, in the vast majority of impotent patients, is sufficient to guarantee its relevance as a therapeutic target. As with numerous other smooth muscle cell types, ion channels are important modulators of corporal smooth muscle tone/contractility. As such, the transmembrane flow of ions (ie Ca(2+), K(+) and Cl(-)) plays an important role in modulating membrane potential and contractile status in individual human corporal smooth muscle cells, while intercellular ion flow ensures the functionality of myocyte cellular networks. The integral membrane proteins that selectively regulate many aspects of these critical transmembrane (eg K(+) and Ca(2+) channels) and intercellular (eg gap junctions) ionic movements have been identified. To date, the large conductance calcium-sensitive K(+) channel (ie K(Ca)), the metabolically regulated K+ channel (ie K(ATP)), and the L-type voltage-dependent Ca(2+) channel appear to be the most physiologically relevant nonjunctional ion channels. With respect to intercellular ionic/solute/second messenger movement, connexin43-derived gap junction channels are widely recognized as an obligatory component to normal integrative erectile biology. The presence of an intercellular pathway ensures that individual cellular alterations are carefully orchestrated in the rapid and syncytial fashion required for normal erectile function. This report reviews the known details concerning junctional and nonjunctional ion channels in human corporal tissue, and illustrates how one particular application of this knowledge, that is, preclinical studies utilizing low efficacy gene therapy (ie low transfection efficiency) with the K(Ca) channel has further confirmed the physiological relevance and therapeutic potential of gap junctions and ion channels to erectile physiology/dysfunction. International Journal of Impotence Research (2000) 12, Suppl 4, S15-S25. PMID- 11035383 TI - Neurotransmitters: central and peripheral mechanisms. AB - Reflexive erection initiated by recruitment of penile afferents, involves both autonomic and somatic efferents. The reflex is mediated at the spinal cord level, modulated by supraspinal influences, and may use several transmitters. Dopamine, acetylcholine, nitric oxide, and peptides, such as oxytocin and ACTH/alpha-MSH, seem to have a facilitatory role, whereas serotonin may be either facilitatory or inhibitory, and enkephalins are inhibitory. Peripherally, the balance between contractant and relaxant factors controls the degree of contraction of the smooth muscle of the corpora cavernosa, and determines the functional state of the penis. Noradrenaline contracts both corpus cavernosum and penile vessels via stimulation of alpha1-adrenoceptors. The role of endothelins in the control of penile smooth muscle tone is presently unclear. Neurogenic nitric oxide (NO) is considered the most important factor for relaxation of penile vessels and corpus cavernosum. The role of other mediators, released from nerves or endothelium has not been definitely established. International Journal of Impotence Research (2000) 12, Suppl 4, S26-S33. PMID- 11035384 TI - Molecular mechanisms for the regulation of penile smooth muscle contractility. AB - Relaxation of smooth muscle is viewed as a 'resetting' of contractile machinery and the resumption of a pre-contractile state is accomplished by lowering cytosolic Ca(2+) and/or by decreasing the sensitivity of the contractile machinery to Ca(2+). There are several mechanisms whereby cytosolic Ca(2+) can be reduced and relaxation achieved but, in general, all pathways depend upon the accumulation of cyclic nucleotides cAMP and cGMP or on the activation of K(+) channels resulting in hyperpolarization. Recently, activation of Na(+)/K(+) ATPase by nitric oxide has been shown to be involved in the relaxation of trabecular smooth muscle. Since Na(+)/K(+) ATPase is electrogenic, its stimulation would cause hyperpolarization and, in turn, would prevent the opening of voltage-dependent Ca(2+) channels. This manuscript briefly reviews the molecular mechanisms affected by muscle relaxants and vasodilators in the treatment of erectile dysfunction. International Journal of Impotence Research (2000) 12, Suppl 4, S34-S38. PMID- 11035385 TI - Pathophysiology of erectile dysfunction: the contributions of trabecular structure to function and the role of functional antagonism. AB - Erectile dysfunction (ED) is estimated to impact more than 150 million men this year worldwide. An understanding of the pathophysiology of ED both furthers the basic scientific knowledge of disease processes and provides a rational design of pharmacotherapy. At present, there are two major views regarding the pathophysiology of ED. In the first hypothesis, the oxygen tension-dependent changes in the penis during erection are proposed to impact corpus cavernosum structure by inducing various cytokines, vasoactive factors and growth factors at the two different oxygen tensions (flaccidity and erection) which, in turn, alter smooth muscle metabolism and connective tissue synthesis. Decreases in the corpus cavernosum smooth muscle/connective tissue ratio have been correlated with an increased likelihood of diffuse venous leak and a failure of the veno-occlusive mechanism in prospective patient studies. Evidence for such a hypothesis incorporates nocturnal penile tumescence and circadian changes in oxygenation as important in maintaining erectile health. The alternate hypothesis proposes that ED is the result of a metabolic imbalance between relaxatory and contractile processes within the trabecular smooth muscle such that contractile processes predominate. Based on this hypothesis, therapy can be accomplished via drugs which shift this balance towards vasodilatation, or by gene therapy approaches to supplement the deficient components favoring smooth muscle relaxation. Both of these hypotheses predict a management strategy for ED that impacts pharmacotherapeutics. In this review of the pathophysiology of ED, each hypothesis will be examined and a synthesis devised incorporating both views. The future of research in this area as well as pharmacotherapy in ED in terms of pathophysiology is discussed including the merits and drawbacks of prophylaxis and prevention of ED. International Journal of Impotence Research (2000) 12, Suppl 4, S39-S46. PMID- 11035386 TI - Medical and surgical advances in the radical prostatectomy patient. AB - Maintaining the quality of life after surgery in the radical prostatectomy patient is of paramount importance. One of the major dilemmas in surgical management of radical retropubic prostatectomy (RRP) is preservation of the neurovascular bundle and, hence, erectile function and the continence mechanism. This manuscript addresses anatomical considerations for the surgeon and discusses the following issues with regard to medical and surgical therapies: (1) incidence of erectile dysfunction solely due to complications during RRP; (2) nerve damage during RRP; (3) vascular damage during RRP; (4) current medical and surgical therapies for restoring or maintaining potency; and (5) new advances on the horizon for management of the prostatectomy patient. International Journal of Impotence Research (2000) 12, Suppl 4, S47-S52. PMID- 11035387 TI - Clinical trials design: protocols and endpoints. AB - Over the last decade there has been a proliferation in clinical trials to test agents for the treatment of erectile dysfunction (ED). Many aspects of clinical trials design and conduct and guidelines for future conduct have been the subject of a recent comprehensive review (Rosen R et al. In: Proceedings of the 1st International Consultation on Erectile Dysfunction 1999). The present article attempts to extend that analysis from trials that focus purely on symptomatic improvement of ED to trials relevant to the management of the ED patient in the community. Although the regulatory approval process accounts for the bulk of the clinical trials undertaken, studies are also initiated for concept testing, post marketing surveillance and for promotional and/or pricing reasons. The trial design can be dependent on which of the above objectives is being served. However, there are also many common features that are summarized below; the major focus is placed on regulatory-standard or 'pivotal' studies. International Journal of Impotence Research (2000) 12, Suppl 4, S53-S58. PMID- 11035388 TI - Opportunities and challenges in oral therapy. AB - The major event in normal erectile function is relaxation of the smooth muscle both within the penile arterial system and the cavernosal tissue. The cavernosal smooth muscle is indistinguishable from peripheral vascular smooth muscle from a physiological point of view, ie both are mediated by the NO-cGMP pathway. In man, the major reason for erectile dysfunction (ED) is 'abnormal smooth muscle relaxation', where the smooth muscle of the cavernosa fails to trap blood within the cavernosa (trapping requires a high intracorporeal pressure). The failure of the cavernosal muscle to relax may be due to inadequate stimulation of the muscle or simply non-compliance of the smooth muscle itself. Therefore, attempts to reverse this abnormal smooth muscle relaxation should be directed at either increasing stimulation to the muscle and/or reversing the poor compliance of the muscle tissue itself. Because the major and most efficient pathway for smooth muscle relaxation is the NO-cGMP pathway, any medication that affects this system should prove to be more potent than other alternate pathways of corporal smooth muscle relaxation, as is evidenced from the clinical observations with current intracorporeal instilled drugs. Because the peripheral vascular smooth muscle is indistinguishable physiologically from the muscle of the cavernosal system, any oral drug must be able to differentiate the two smooth muscle compartments to prevent systemic side effects that could render such drugs clinically useless. At the present time, there are no methods to improve smooth muscle compliance but advances in tissue engineering may allow us to accomplish this in the future. International Journal of Impotence Research (2000) 12, Suppl 4, S59-S61. PMID- 11035389 TI - Sildenafil citrate: current clinical experience. AB - There is little question as to the efficacy of sildenafil citrate (Viagra) for the treatment of erectile dysfunction. Two years of post-marketing experience provide data which indicates that the efficacy and safety of sildenafil citrate is consistent with the data obtained in clinical trials. This paper provides an update of the clinical efficacy and safety of sildenafil collected since its market approval in March of 1998. International Journal of Impotence Research (2000) 12, Suppl 4, S62-S66. PMID- 11035390 TI - Apomorphine: an update of clinical trial results. AB - Apomorphine SL (TAP Holdings, Deerfield, IL) is a centrally acting treatment for erectile dysfunction (ED) that has been undergoing phase III trials. Over 3000 men have received apomorphine SL and over 75,000 doses have been taken. In the first three phase III parallel arm cross-over double-blind studies 854 patients were given a total of 8263 tablets of apomorphine SL in 2 and 4 mg doses. The patients were between 18 and 70 y old and outcome measures included per attempt rates of intercourse and erections firm enough for intercourse as well as psychometric instruments and partner responses. The majority (74.1%) had moderate and severe grades of ED on admission to the studies, 31% had hypertension, 16% had documented coronary artery disease, 16% had dyslipidemia and 16% had diabetes. Erections occurred rapidly (10-25 min) and in 54.4% of attempts at 4 mg (vs 33.8% placebo). A majority of the attempts at intercourse (50.6%) were successful at 4 mg in patients when recorded on a per-attempt basis. The most common but infrequent and mild side effect of nausea decreases with use. The phase III trials of apomorphine SL show that there is a clinically important restoration of erectile function from this new formulation of apomorphine. It has a rapid and safe effect through action in the central nervous system. Apomorphine SL brings a new choice to the management of ED that will further benefit the millions of couples affected. International Journal of Impotence Research (2000) 12, Suppl 4, S67-S73. PMID- 11035391 TI - Melanocortin receptor agonists, penile erection, and sexual motivation: human studies with Melanotan II. AB - We review our experience with Melanotan II, a non-selective melanocortin receptor agonist, in human subjects with erectile dysfunction (ED). Melanotan II was administered to 20 men with psychogenic and organic ED using a double-blind placebo-controlled crossover design. Penile rigidity was monitored for 6 h using RigiScan. Level of sexual desire and side effects were reported with a questionnaire. In the absence of sexual stimulation, Melanotan II led to penile erection in 17 of 20 men. Subjects experienced a mean of 41 min Rigiscan tip rigidity>80%. Increased sexual desire was reported after 13/19 (68%) doses of Melanotan II vs 4/21 (19%) of placebo (P<0.01). Nausea and yawning were frequently reported side effects due to Melanotan II; at a dose of 0.025 mg/kg, 12.9% of subjects had severe nausea. We conclude that Melanotan II is a potent initiator of penile erection in men with erectile dysfunction. Our findings warrant further investigation of melanocortin agonists and antagonists on penile erection. International Journal of Impotence Research (2000) 12, Suppl 4, S74 S79. PMID- 11035392 TI - Developmental status of topical therapies for erectile and ejaculatory dysfunction. AB - The most common of the sexual dysfunctions in men are premature ejaculation and impotence. Large strides have been made in the treatment of erectile disturbances but only limited and temporary successes have been achieved in the therapy of ejaculatory abnormalities. This manuscript examines the realm of topical therapy, its current limitations and the challenges that must be addressed if topical therapy is ever to have a niche in the urologist's treatment armamentarium. Topical agents for the treatment of premature ejaculation include anesthetics and herbal medications. The limited nature of the studies reported to date does not yet permit a reliable assessment of their efficacy. For the treatment of erectile dysfunction, some of the drugs administered by various other systemic routes appear to have some efficacy when delivered transdermally. Again, the studies and results are too limited for the urologist to develop a clear opinion about their efficacy. Further investigation of these drugs with the use of absorption enhancers is needed. International Journal of Impotence Research (2000) 12, Suppl 4, S80-S85. PMID- 11035393 TI - Review of intraurethral suppositories and iontophoresis therapy for erectile dysfunction. AB - This brief review examines the history of the intraurethral (IU) pharmacotherapy for the treatment of erectile dysfunction (ED). Emphasis is placed on the design, study endpoints and results of a limited number of clinical trials undertaken with PGE(1)-MUSE to demonstrate the efficacy of this unique delivery system. Next, the theory of the iontophoretic technique of drug delivery is discussed. Clinical data collected while applying these methods are presented, but are thus far limited to treatment of patients with Peyronie's disease. Iontophoresis as a drug delivery alternative may gain popularity with ED patients who fail other types of therapy or in patients with other penile disorders. International Journal of Impotence Research (2000) 12, Suppl 4, S86-S90. PMID- 11035394 TI - Current perspectives on intracavernosal pharmacotherapy for erectile dysfunction. AB - Thirty percent of males afflicted with erectile dysfunction (ED) do not respond to oral drugs, another 15% reveal contraindications to currently available therapy, and a subset of patients actually prefer injection therapy due to its predictable short time-to-onset of erection and reliable rigidity compared to oral drugs. This paper provides both a historical and current perspective on intracavernosal therapy and reviews the popular injectable therapeutic agents that are currently used for the treatment of ED. Emphasis is placed on the efficacy, mechanism of action and side effect profiles of approved and experimental injectable pharmacotherapy for ED. International Journal of Impotence Research (2000) 12, Suppl 4, S91-S100. PMID- 11035395 TI - Historical advances in penile prostheses. AB - In the 25 y since their introduction, semi-rigid and inflatable penile implants have become remarkably dependable mechanical instruments associated with high patient satisfaction. This report attempts to quantify the historical milestones of significance pertaining to these devices. As with any historical 'best of ' list, there will be controversy and omissions. Three broad topics contributing to the advancement of penile prosthetic surgery, devices and techniques are discussed: (1) Prosthetic design changes contributing to freedom from revision: distention controlled cylinders, Bioflex cylinders, connectionless systems and reservoir lockout valves; (2) Instrument innovations to facilitate prosthetic surgery: Scott retractor, Furlow inserter, Brooks dilators, Carrion-Rossello cavernotomes; (3) Clever surgical applications: SST repair, transverse scrotal incision, modeling for Peyronie's disease, salvage for infection and natural tissue repair. International Journal of Impotence Research (2000) 12, Suppl 4, S101-S107. PMID- 11035396 TI - Surgical management of penile prosthesis complications. AB - Penile implants are mechanical devices used to treat erectile dysfunction. Parts of the implants may wear out with time, and the implants themselves may damage the body cavities in which they have been placed, especially with excessive or aggressive use. Techniques to replace parts, repair body cavities, and deal with infection associated with prosthesis placement have been developed. Satisfaction with the resulting erections is the highest among all the modalities available for treating erectile dysfunction. International Journal of Impotence Research (2000) 12, Suppl 4, S108-S111. PMID- 11035397 TI - Testosterone replacement: when is there a role? AB - Hypogonadism is an uncommon cause of erectile dysfunction. Unfortunately, hypogonadal states in adult males are difficult to diagnose on purely clinical grounds and it is necessary to seek biochemical support. The simplest way to establish the diagnosis of hypogonadism is by determination of serum testosterone levels. Several methods exist but total testosterone determination plus assessment of sex hormone-binding globulin or bio-available testosterone appear to be the most reliable and accessible. Once a diagnostic of hypogonadism has been established in a man with erectile difficulties, a trial of androgen supplementation is warranted if no contraindications exist. Knowledgeable monitoring is essential. In the absence of an adequate response, co-morbidities should be diligently sought out. In the absence of reliable guidelines for androgen administration to patients with erectile failure, a set of recommendations are provided. International Journal of Impotence Research (2000) 12, Suppl 4, S112-S118. PMID- 11035398 TI - Diagnostic and treatment strategies for erectile dysfunction: the 'Process of Care' model. [University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School]. AB - The new era of erectile dysfunction medicine ushered in by the availability of an effective and safe oral medication paves the evolution of the field toward a multi-disciplinary and primary care setting. There exists a paucity of guidelines regarding the assessment and management of the patient with erectile dysfunction in the primary care arena. The 'Process of Care' model for the evaluation and treatment of erectile dysfunction has been developed to advance new guidelines for the diagnosis and management of erectile dysfunction in the primary care and multi-disciplinary setting. This model was developed under the auspices of the University of Medicine and Dentistry of New Jersey (UMDNJ)-Robert Wood Johnson Medical School (chairman: R Rosen) with input from a multi-disciplinary group of experts including representation from primary care, internal medicine, endocrinology, psychiatry, psychology and urology. The methodology employed was a modified delphi-technique (RAND method). The key components of the model are: 1)a rational approach to diagnosis and treatment, 2) emphasis on clinical history taking and a focused physical examination 3) specialized testing and referral in pre-defined situations 4) step-wise management approach with ranking of treatment options and 5)incorporation of patient and partner needs and preferences in the decision making process when possible (a goal-directed approach). The management algorithm is conceptually organized into progressive stages from Process--> Action--> Outcome. International Journal of Impotence Research (2000) 12, Suppl 4, S119-S121. PMID- 11035399 TI - Penile prosthesis implantation in the treatment of Peyronie's disease and erectile dysfunction. AB - Although men afflicted with Peyronie's disease (PD) usually have a number of treatment options, those who also present with erectile dysfunction (ED) arising from unknown or iatrogenic causes are not easily treated. Surgical straightening procedures that have been used to treat PD may not restore erectile function and failure to straighten the penis with surgery may be the result of erectile inadequacy during the post-operative period. This paper discusses penile prosthesis implantation as a surgical option for patients with PD, placing emphasis on the choice of devices and surgical techniques. Several new techniques which hold the promise of high success rates and low morbidity are mentioned. International Journal of Impotence Research (2000) 12, Suppl 4, S122-S126. PMID- 11035400 TI - The genetics and immunology of Peyronie's disease. AB - Peyronie's disease (PD) is a condition characterized by localized and often progressive fibrosis and scarring of the penis. This condition has an unknown etiology although several hypotheses have been proposed. These include traumatic, immunologic and genetic causes. We studied the genetics and immunology of PD using both molecular biologic and molecular genetic techniques. Men (n=283) with PD were identified by retrospective chart review of one physician's office practice. These men were contacted by telephone and asked to submit to an interview and blood test for genetic studies. Simultaneously, tissue and cells collected in the laboratory were examined by Western and Northern blot analysis for examination of protein and RNA for expression of HLA. Of the first 107 men contacted, 24 were available and consented to interview and blood testing. The mean age was 60.3 y with an average duration of PD of 4.9 y. One patient had a family history of PD while no patients had Dupuytren's contracture. Twenty patients were considered to have primary disease while four were secondary. Eleven patients had tissue prepared for Northern blot analysis and nine patients were the subject of Western blot analysis. All tissue, both Peyronie's and control expressed class I MHC while no tissue expressed class II MHC. The expression of mRNA of class I MHC was equal for Peyronie's and control patients while the expression at the protein level was less in the PD patients. We conclude that PD may have multiple etiologic agents. One cannot exclude a class II MHC association but in our population, HLA DQ is not expressed. Class I MHC may be involved as the expression of class I MHC protein is different in Peyronie's patients than in controls. Genetic studies are ongoing. International Journal of Impotence Research (2000) 12, Suppl 4, S127-S132. PMID- 11035401 TI - Priapism. AB - Priapism is a prolonged, painful, penile erection that fails to subside despite orgasm. An erection lasting longer than 4-6 h is considered to be priapic; nevertheless, pain does not usually ensue until 6-8 h have elapsed. Priapism is considered a failure of the detumescence mechanism, which may be due to excess release of contractile neurotransmitters, obstruction of draining venules, malfunction of the intrinsic detumescence mechanism, or prolonged relaxation of intracavernosal smooth muscle. There are essentially two main types of priapism: high flow (non-ischemic) and low flow (ischemic). Low flow priapism is the more common form, and it is associated with a decrease in venous outflow and vascular stasis that, in turn, cause tissue hypoxia and acidosis. This form of priapism is usually quite painful because of tissue ischemia. Penile blood aspirated from cavernous spaces appears dark in color. Immediate treatment is necessary or penile fibrosis will ensue. High flow priapism is usually due to trauma, although, on rare occasions it has been idiopathic or due to sickle cell disease. The hallmark of this type of priapism is an increase in arterial inflow in the setting of normal venous outflow. Aspirated penile blood is noted to be bright red and has a high pO(2). This form of priapism is not usually painful because it is non-ischemic. Treatment is dependent on the wishes of the patient but is not mandatory. International Journal of Impotence Research (2000) 12, Suppl 4, S133 S139. PMID- 11035402 TI - Future considerations: advances in the surgical management of erectile dysfunction. AB - As treatment options for erectile dysfunction (ED) continue to expand, and with more attractive alternatives such as effective systemic treatment becoming available, the number of men presenting for treatment of ED is increasing exponentially. Since a subset of these men continue to require surgical therapy, there is a potential for the number of operations for the treatment of ED and related disorders to actually increase. Areas in the surgical treatment of ED where improvements are needed are identified, including: measures to prevent penile prosthesis infections, better penile implants, improved penile augmentation procedures, better surgical procedures for the treatment of Peyronie's disease, improved penile revascularization procedures, and new motor and sensory penile nerve grafting procedures. International Journal of Impotence Research (2000) 12, Suppl 4, S140-S143. PMID- 11035403 TI - Impact of erectile dysfunction on quality of life: patient and partner perspectives. AB - Quality of life (QoL) has become one of the important parameters in the evaluation of treatment and assessment of medical conditions, and it may be an important tool in determining the urgency of the need for therapeutic intervention for erectile dysfunction (ED). It is important to evaluate QoL of the couple, because men and women alike will suffer because of male erectile disability. Future drug trials, as well as studies of sexological intervention programs, should involve both partners. International Journal of Impotence Research (2000) 12, Suppl 4, S144-S146. PMID- 11035404 TI - Screening for erectile dysfunction: rationale. AB - A number of epidemiologic and clinical studies that have examined the relationship between erectile dysfunction, depression and cardiovascular disease are reviewed. Hypothetical mechanisms that can explain the correlations observed among these phenomena are described and a mutually reinforcing predictive model is proposed. International Journal of Impotence Research (2000) 12, Suppl 4, S147 S151. PMID- 11035405 TI - Female sexual arousal disorder: new insights. AB - Epidemiologic investigations of women with female sexual dysfunction (FSD) from well-designed, random-sample, community-based populations are limited. Based on available information, FSD is common and estimated to occur in 22-43% of women. There are limited data on age-related and para-aging risk factors, which are critical to understand when planning treatment and prevention efforts. Based on correlates of FSD, associated risk factors include age, education, history of sexual abuse or sexually transmitted disease, overall state general happiness and physical health. This brief overview attempts to review what is known about the female sexual anatomy, describes factors that may affect female sexual responsiveness, and identifies several areas where additional research is needed to promote understanding of this complex physiological and psychosocial phenomenon. International Journal of Impotence Research (2000) 12, Suppl 4, S152 S157. PMID- 11035406 TI - New therapies and delivery mechanisms for treatment of erectile dysfunction. AB - Despite the successes of Viagra, the quest for new and better therapy for erectile dysfunction (ED) continues. In a recent survey of the first 220 patients placed on Viagra at our institution, 101 (46%) quit taking the drug: 76% of those who quit were not satisfied with the results. Patients clearly want an efficacious, safe, convenient medication with rapid onset. To meet these consumer demands, numerous new therapies are being developed. These include new oral medications, new intracavernosal pharmacotherapies, new delivery systems (such as novel intracorporal injectors and transdermal agents) and combination therapies. What is known about these new medications and delivery systems will be presented. Hopefully, from these innovations will come therapies that will improve the overall success and acceptance of treatment for ED. Since it is unlikely that any single agent will ever provide a solution for all men with ED, an expanded armamentarium of treatment options will greatly enhance the chances that any given man will be able to find a therapy that is both acceptable and appropriate to him. International Journal of Impotence Research (2000) 12, Suppl 4, S158 S162. PMID- 11035407 TI - Penile erectile function: recommendations for future research. AB - Research in the field of erectile function and dysfunction is rapidly expanding. Future directions of this research were discussed at the 1st International Consultation on Erectile Dysfunction, held in Paris, July 1999. Important research areas were identified, and some recommendations were made. There is a need for mouse models of erection for molecular biology purposes, and also for new animal models of erectile dysfunction. Behavioral assessments of sexual function are advantageous and deserve to be used more widely. More knowledge about the anatomical interrelations and interactions between different brain nuclei that modulate erection and sexual function is required. Centrally and peripherally, the potential of new neuronal and non-neuronal transmitters and modulators and their interactions should be further explored. The signal pathways in erectile tissues for different transmitters/modulators should be studied in detail, and also the distribution and roles of eg, receptors, cyclases, phosphodiesterases, and protein kinases. Further information is desired about excitation-contraction coupling, cell-to-cell transmission of activation, ion channels within erectile tissues and also about changes in contractile proteins that occur with erectile dysfunction. Important for treatment are strategies to prevent cavernosal degeneration and/or to restore cavernosal function. Intracavernosal somatic gene therapy is promising, and new molecular targets for drug treatments should be identified and explored. International Journal of Impotence Research (2000) 12, Suppl 4, S163-S167. PMID- 11035409 TI - Erratum. PMID- 11035408 TI - The first international conference on the management of erectile dysfunction conference participants PMID- 11035410 TI - 3rd International Symposium on NeuroVirology. PMID- 11035411 TI - Eppur si muove (Galileo galilei 1564-1642): the idiotypic dysregulation of autoantibodies as part of the etiology of SLE. PMID- 11035412 TI - 'Catastrophic' antiphospholipid syndrome. AB - When 'catastrophic' is applied as an adjective to the antiphospholipid syndrome, it implies a characteristic presentation due to predominantly small blood vessel thrombosis leading to rapidly progressive failure of multiple organs and a frequently fatal outcome. We present the case of a 48-year-old woman who presented with the 'catastrophic' antiphospholipid syndrome without previous history of coagulation disorder or connective tissue disease that illustrates the difficulties in diagnosing and managing this disorder. We also review the factors that have been reported to have a role in the development of this condition and show how this case throws light on its pathogenesis. PMID- 11035413 TI - Induction of anti-DNA antibodies by immunization with anti-DNA antibodies: mechanism and characterization. AB - Two well-characterized IgG monoclonal antibodies, reactive with double-stranded (ds) DNA and nucleosomes, were administered to normal BALB/c mice to examine the reproducibility and the biology of a previously reported model of anti-DNA antibody induction by immunization with anti-DNA antibodies. The monoclonal antibodies were purified either with or without a high-salt wash to remove nucleosomal antigens bound to them during the cell culture. Both monoclonal antibodies, but not normal IgG, induced significant IgG anti-dsDNA antibody production from 1 week to 25 weeks after the last immunization. The antibodies produced in this manner possess different binding preferences to ds synthetic polynucleotides than the antibodies used for the immunization, and they did not react with nucleosomes. The monoclonal antibodies purified with the high-salt wash were more effective in anti-DNA antibody induction than those purified without the high-salt wash. Even when bound to these monoclonal antibodies, neither dsDNA, nucleosomes, or ds synthetic polynucleotides exert significant antigenicity. For example, anti-DNA antibodies produced by mice immunized with an immune complex formed by poly(dA-dT) and one of the monoclonal antibodies that has a high affinity to this polynucleotide did not show an increased affinity to poly(dA-dT). Together, these results suggest that anti-DNA antibody molecules or processed antibody peptides, and not DNA/nucleosomes carried by anti-DNA antibodies, play a role in this model of anti-DNA antibody production. PMID- 11035414 TI - Intrathecal cytokines in systemic lupus erythematosus with central nervous system involvement. AB - Symptoms originating from central nervous system (CNS) are frequently occuring in patients with systemic lupus erythematosus (SLE). Reliable diagnostic markers for this condition are presently lacking. Importantly, CNS involvement in lupus patients is associated with increased morbidity and mortality. The aim of this retrospective evaluate was to study the diagnostic value of cerebrospinal fluid (CSF) cytokine levels in SLE patients with CNS involvement. 34 patients with SLE were hospitalized and investigated for the presence of CNS lupus. These patients were evaluated clinically and with magnetic resonance imaging (MRI) and CSF analyses, as well as with neuropsychiatric tests. 13 patients were found to have CNS lupus whereas another four of the patients fulfilled the criteria for CNS involvement but were excluded from this group due to other causes of CNS involvement. Lastly, in 17 SLE cases, the diagnosis of CNS lupus could not be confirmed. CSF levels of interleukin-6 (IL-6) and IL-8, as well as the CSF/serum IL-6 ratio, were elevated in the CNS lupus group, compared with the 17 SLE patients not fullfilling a diagnosis of cerebral lupus. Interestingly, follow-up of five patients being successfully treated for CNS lupus revealed profound decrease of intrathecal IL-6 levels. These results indicate that analysis of CSF cytokine levels, especially IL-6 and IL-8, may be useful in the diagnostics and possibly follow-up of SLE patients with cerebral lupus. PMID- 11035415 TI - Renal immunofluorescence and the prediction of renal outcome in patients with proliferative lupus nephritis. AB - The risk for endstage renal failure in patients with proliferative lupus nephritis (PLN) depends largely on the severity and reversibility of the inflammatory process as determined by light microscopy (LM). As the intrarenal formation of immune complexes is thought to initiate this inflammation, we studied whether renal immunofluorescence microscopy (IFM) provides clinical or prognostic information in addition to LM findings. Clinical data at the time of renal biopsy and during a mean follow-up of 46 months were extracted from the records of 69 SLE patients with proliferative LN (WHO class III/IV). Biopsy specimens were analyzed by LM for AI and CI, while IFM was performed on cryostat sections with the use of antisera against IgG, IgM, IgA, C3, C1q and fibrin. IFM findings were recorded in terms of the localization (glomerular, tubular or vascular) and intensity of fluorescence (score from zero to three). IFM findings were then related to clinical and LM findings and its prognostic value studied by survival analysis. Glomerular immune deposits were present in 99% of patients, tubular deposits in 38% and vascular deposits in 17%. A 'full-house' pattern (all three Ig classes) was found in 67% of biopsies and C3 and C1q deposits in 93% and 74% respectively. Median scores for AI and CI were 6 (1-18) and 3 (0-10); aside from a negative correlation between IgA deposits and CI, we found no other correlation between the amount or type of immune deposits and AI or CI. IgM deposits were associated with high serum levels of anti-dsDNA, while IgG deposits correlated with high ESR and serum creatinin levels. IFM scores were not related to steroid dose at the time of biopsy and neither type of glomerular, tubular or overall renal immune deposits had prognostic value for renal survival. Renal immunofluorescence does not reflect light microscopy findings in patients with PLN and does not contribute prognostic information in patients with PLN. Lupus (2000) 9, 504-510. PMID- 11035416 TI - The spectrum of ocular involvement in patients with systemic lupus erythematosus without ocular symptoms. AB - The aim of the study was to determine the spectrum of clinical ocular involvement in patients with inactive systemic lupus erythematosus (SLE) who have no ocular symptoms. Patients with a diagnosis of SLE based on the 1982 revised American College of Rheumatology criteria and with no ocular complaints were recruited from the SLE clinic. Clinical data regarding their systemic disease and disease activity were recorded and a full ophthalmic examination carried out. 52 patients of mixed ethnicity comprising of 75% Chinese, 19% Malays and 6% Indian patients were recruited. Of these, 51 (98%) were female with a mean age of 34+/-11 (range 16-74 y). 16 (31%) patients had dry eyes while corticosteroid induced glaucoma and cataract was detected in 1 (2%) and 7 (14%) patients, respectively. No patients were found to have sight-threatening ocular conditions such as cotton wool spots, vasculitis, optic neuropathy or uveitis. Patients with clinically inactive disease were found not to have sight-threatening ocular diseases that are known to be associated with SLE. Although they have no ocular complaints, nearly one-third of these patients have dry eyes. Ocular examination may be unnecessary when the disease is clinically inactive and in the absence of ocular symptoms. PMID- 11035417 TI - Determination of autoantibodies to annexin XI in systemic autoimmune diseases. AB - Annexin XI, a calcyclin-associated protein, has been shown to be identical to a 56,000 Da antigen recognized by antibodies found in sera from patients suffering from systemic autoimmune diseases. In this work hexahistidine-tagged recombinant annexin XI (His6- rAnn XI) was used as antigen in ELISA experiments for determination of autoantibodies to annexin XI in sera of patients with systemic rheumatic autoimmune diseases. Immunoblotting with HeLa cell extract and with His6-rAnn XI as antigen was used for confirmation of positive ELISA results. We found eleven anti-annexin XI positive sera (3.9%) out of 282 sera from patients with systemic rheumatic diseases. The highest number of annexin XI positive sera were found in primary antiphospholipid syndrome (3/17), and in subacute lupus erythematosus (1/6), while lower frequencies of positive sera were found in patients with systemic sclerosis (5/137), rheumatoid arthritis (1/21), and systemic lupus erythematosus (1/58). Sera from healthy donors and patients with chronic infections were negative, except for one Salmonella typhimurium antibody positive serum. Autoantibodies to annexin XI were found to relate to thrombosis, but not to other clinical or laboratory features. A relation between antibodies to annexins and thrombosis has so far only been known for annexin V. PMID- 11035418 TI - Subarachnoid hemorrhage and systemic lupus erythematosus. AB - The frequency, clinical profile, treatment and outcome of subarachnoid hemorrhage (SAH) in patients with systemic lupus erythematosus (SLE) were assessed retrospectively, based on the case records of SLE of the Jichi Medical School Hospital over a 20 year period. Clinically defined SAH was found in 10 (3.9%) out of 258 SLE patients, which represented a frequency higher than previously assumed. Five patients had active SLE and lacked an apparent cause of SAH, other than SLE. A high mortality rate (5/5), no visible aneurysm on angiogram (3/4), and an onset during intractable SLE or after discontinued or no steroid therapy because of medical noncompliance (4/5) were characteristic of patients with active SLE, and thus an earlier successful suppression of SLE, if possible, might have prevented their SAH. In contrast, in the 5 patients with inactive SLE, 2 out of 3 saccular aneurysms were successfully clipped and small bleeding of one patient without aneurysms remitted spontaneously without the need for additional steroid therapy. When one death, which occurred outside of medical care, was excluded, the survival ratio of the hospitalized SAH patients with inactive SLE was significantly better than that with active SLE (3/4 versus 0/5, P=0.0476). In conclusion, the relatively common occurrence of SAH in SLE patients, and a significantly different clinical impact of SAH in respect to active and inactive SLE, were suggested from the results. PMID- 11035419 TI - Mood states and disease activity in patients with systemic lupus erythematosus treated with bromocriptine. AB - We tested mood states in patients with systemic lupus erythematosus (SLE) treated with the prolactin-lowering drug, bromocriptine. Bromocriptine was given to seven patients in an open-label study to test its effects on active SLE. Two independent measures of SLE activity, the SLE Activity Measure (SLAM) and the SLE Disease Activity Index (SLEDAI), were scored and the Symptom Questionnaire (SQ) mood survey was administered at entry and at 6 monthly follow-up visits. The SLAM and SLEDAI scores improved significantly during treatment. Two of the four mood scales in the SQ (Anxiety Scale and Anger-Hostility Scale) showed significant improvement compared to the entry value at least once during treatment. Significant improvement was also observed in the Total Distress Score, which is the sum of the four scales and is a more sensitive measure of distress than the score of an individual scale. Depression, anxiety, somatic complaints, and total distress correlated positively with SLAM and/or SLEDAI scores. The Anxiety Scale and the Total Distress Score improved with treatment and did correlate positively with SLE activity. In contrast, the Anger-Hostility Scale improved with treatment but did not correlate with SLE activity. PMID- 11035420 TI - Illness intrusiveness explains race-related quality-of-life differences among women with systemic lupus erythematosus. AB - Our objective was to investigate whether quality of life in systemic lupus erythematosus (SLE) differs across ethnoracial groups and to identify factors that may explain race-related differences. Self-administered questionnaire data from 335 White, 40 Black, and 30 Asian women with SLE were obtained from a multi center database. Measures assessed illness intrusiveness, psychological well being, depressive symptoms, musculoskeletal pain, and learned helplessness. Extent of SLE disease activity was indexed by self-reported functional-system involvement. Educational attainment was indicated by number of years in school. Principal-components analysis reduced the four psychosocial measures to a single factor score. This represented psychosocial well-being In path analysis. Psychosocial well-being differed significantly across the three groups, with Whites reporting the highest, and Blacks the lowest, levels. Path analysis indicated that illness intrusiveness accounted for this race-related difference. Although disease activity was significantly associated with psychosocial well being, it did not differ across ethnoracial groups. Illness intrusiveness and educational attainment emerged as independent mediators of the race-related difference in psychosocial well-being. We conclude that race-related quality-of life differences exist among women with SLE and are mediated independently by illness intrusiveness and educational attainment. PMID- 11035421 TI - Increased cervical dysplasia in intravenous cyclophosphamide-treated patients with SLE: a preliminary study. AB - To determine if intravenous cyclophosphamide (IV-C) causes an excess of cervical dysplasia and/or cancer in systemic lupus erythematosus (SLE) patients, a retrospective review was conducted. Patients with SLE who received IV-C between 1988-98 (study group) were compared with a group of SLE patients who had not received IV-C (control group). Of the 79 IV-C-treated SLE patients identified, we excluded 18 because of absence of pertinent data. We found 10 cases of cervical dysplasia in the remaining 61 patients, compared to 2 in 49 non-exposed patients (P<0.04). Comparison of the two groups revealed no difference in: mean years of disease duration, months of follow-up and age. The non-exposed patients were more likely to be on estrogen and hydroxychloroquine but less often on steroids and azathioprine. The study group with and without dysplasia were assessed; we found no difference in the mean, or total IV-C dose, smoking and estrogen use. There was a significant decrease in time to dysplasia in those, given IV-C, with previous dysplasia compared to those without. These preliminary data suggests that IV-C causes an increased number of abnormal Papanicolaou (Pap) smears in SLE patients, particularly those with previous dysplasia. PMID- 11035422 TI - Anetoderma in systemic lupus erythematosus: relationship to antiphospholipid antibodies. AB - Anetoderma is an elastolytic disorder where multiple patches of slack skin are formed. Twelve patients with anetoderma associated with systemic lupus erythematous have been described, all in the dermatological literature. Recently, a role for antiphospholipid antibodies has been proposed with microthromboses as its pathogenic mechanism. We present herein a 20-year-old female patient who developed anetoderma soon after sun exposure. She was found to have a false positive VDRL and gradually developed other manifestations of SLE, including interstitial cystitis. She has had repeatedly positive antiphospholipid antibodies. Although there are patients who may have a primary form, diagnosis of anetoderma should trigger a search for SLE and/or antiphospholipid antibodies. PMID- 11035423 TI - Crossing of antinuclear antibodies and anti-leishmania antibodies. AB - Visceral leishmaniasis or Kala-azar is an endemic parasitic infection in Mediterranean countries. We report an interesting case occurring in a 38-year-old woman suffering from systemic lupus erythematosus and secondary antiphospholipid syndrome. In our patient visceral leishmaniasis occurred during high dose steroids treatment mimicking a flare of lupus. As the lupus resolved under immunosuppressive treatment, a reactivation of visceral leishmaniasis was observed and was confirmed by the successive serological tests which showed crossing of leishmania and antinuclear antibody titers. Our case shows that, faced with fever occurring in lupus patients in an endemic area, visceral leishmaniasis should be searched for before intensifying immunosuppressive treatments. PMID- 11035424 TI - Parvovirus B19 infection associated with the production of anti-neutrophil cytoplasmic antibody (ANCA) and anticardiolipin antibody (aCL). AB - We described four patients who had clinical diagnosis of erythema infectiosum and presented with skin rash, polyarthralgia, polyarthritis, and mild fever. Anti parvovirus B19 IgM and IgG antibodies were found in all four patients and parvovirus B19 DNA was detected in three of the four patients by polymerase chain reaction (PCR) in sera using standard methods. Anticardiolipin antibody (aCL) was positive in three of the four patients included three with anti-beta2 glycoprotein I (beta2GPI). The immunoglobulin isotype of aCL was found to be IgM. Anti-neutrophil cytoplasmic antibody (ANCA) included three p-ANCA and one c-ANCA was detected in all four patients by indirect immunofluoresence (IIF). Both anti proteinase 3 (PR3) and anti-myeloperoxidase (MPO) antibodies were found in two patients whom had polyarthritis for more than 6 months. These data indicate parvovirus B19 may be linked to the induction of an autoimmune response. PMID- 11035425 TI - Lamotrigine-induced lupus. AB - A 57-year-old woman was under treatment for epilepsy with lamotrigine 2 mg/kg/d since 1996. In April 1998 she was admitted to the Rheumatology Unit for arthralgias affecting the small joints of the hands, wrists and knees, erythematosus skin rash and myalgias. She referred a vascular abnormality at the hands exposed to cold, compatible with Raynaud's phenomenon. Serum antinuclear antibodies (ANA) were positive (1:320, speckled pattern); moreover, a positivity for anti-Ro/SSA was observed. Rheumatoid factor was negative, serum complement was normal. LAC and anticardiolipin antibodies were negative. Drug-related lupus diagnosis was made with resolution of symptoms and gradual normalisation of positivity of ANA and anti-Ro/SSA upon lamotrigine withdrawal. To our knowledge, this is the first case report of an association between lamotrigine and the onset of SLE (systemic lupus erythematosus). PMID- 11035426 TI - Pulmonary embolism and transitory anti-beta2-GPI antibodies in an adult with chicken pox. AB - Anti-beta2-glycoprotein I antibodies are considered as a specific marker for the antiphospholipid syndrome. In contrast to lupus circulating anticoagulant and anticardiolipin (aCL) antibodies, they are usually not found at significant levels in infections. We report a case of pulmonary embolism in an adult with varicella. Transient significant levels of aCL antibodies and of IgM anti-beta2 GPI antibodies were observed. No other prothrombotic factor, including free protein S antigen deficiency, was found. The direct pathogenic role of these transient antibodies on the thrombotic event may then be suspected. They are probably associated with VZV acute infection and are absent two months after varicella. PMID- 11035427 TI - Should low-dose aspirin also be a background therapy for all patients with systemic lupus erythematosus (SLE)? PMID- 11035428 TI - Cancer & Autoimmunity, by Y Shoenfeld and ME Gershwin. PMID- 11035429 TI - Images of lupus: diving for pearls. PMID- 11035430 TI - Nonsteroidal anti-inflammatory drugs in systemic lupus erythematosus. AB - Up to 80% of patients with systemic lupus erythematosus (SLE) are treated with nonsteroidal anti-inflammatory drugs (NSAID) for musculoskeletal symptoms, serositis and headache. This survey reviews the literature on non-selective and selective inhibitors of cyclooxygenases, with an emphasis on the efficacy and safety profile reported in SLE patients. No lupus-specific data on gastro intestinal side effects of NSAID exist. Both non-selective Cox inhibitors and selective Cox-2 inhibitors induce renal side effects, including sodium retention and reduction of the glomerular filtration rate. Lupus nephritis is a risk factor for NSAID-induced acute renal failure, but not for rare idiosyncratic toxic renal reactions to NSAID. In refractory nephrotic syndrome, NSAID have been used successfully. Cutaneous and allergic reactions to NSAID are increased in SLE patients as well as hepatotoxic effects, particularly with high dose aspirin. Whereas a variety of central nervous system side effects of NSAID are probably no more common in SLE patients than others, aseptic meningitis has been reported more frequently. Ovulation and pregnancy can be adversely affected by Cox inhibitors. The antiplatelet effect of aspirin and non-selective Cox inhibitors has a therapeutic potential in patients with antiphospholipid syndrome (APS). In summary, treatment of SLE with NSAID requires awareness for the increased frequency of some side effects and close monitoring of toxicity. PMID- 11035431 TI - Central nervous system involvement in systemic lupus erythematosus: cerebral imaging and serological profile in patients with and without overt neuropsychiatric manifestations. AB - The aim of this study was to evaluate morphological and functional abnormalities by cerebral imaging in a series of systemic lupus erythematosus (SLE) patients with and without overt central nervous system (CNS) manifestations, and to detect possible relationships with clinical parameters and a large panel of autoantibodies, including those reactive against neurotypic and gliotypic antigens. 68 patients with SLE were investigated in a cross-sectional study which included clinical evaluation of symptoms, cerebral magnetic resonance imaging (MRI) and brain single photon emission tomography (SPECT) analysis, electroencephalography (EEG), and serological tests for antibodies directed against nuclear, cytoplasmic neuronal and glial cell-related antigens. The results of this study showed: (1) a significant positive association of (a) anti glial fibrillary acidic protein (GFAP) serum antibodies with neuropsychiatric (NP) manifestations and (b) anti-serin proteinase 3 (anti-PR3/c-ANCA) serum antibodies with pathological cerebral SPECT; (2) the presence of significantly higher values of (a) SLICC organ damage index in patients with abnormal MRI and (b) SLAM activity index in patients with abnormal SPECT; and (3) the association of (a) abnormal MRI with nonactive NP manifestations and (b) combined abnormality of brain SPECT and MRI with the occurrence of overall overt NP manifestations and with those of the organic/major type. Neuropsychiatric manifestations, namely those of the organic/major type, appeared to be significantly associated to the presence of a serum antibody against GFAP, a gliotypic antigen. There was also evidence of an association between SPECT abnormality and the presence of anti-PR3 (c-ANCA). Furthermore, brain imaging by MRI and SPECT applied to SLE patients appears to express CNS involvement significantly related to specific categories of NP manifestations. The abnormalities detected by the two tests seem to be preferentially associated with different activity phases of the NP disorder or of the lupus disease. PMID- 11035432 TI - Psychiatric and psychosocial disorders in patients with systemic lupus erythematosus: a longitudinal study of active and inactive stages of the disease. AB - The objective was to analyze psychiatric disorders and psychosocial dysfunction in patients with systemic lupus erythematosus (SLE), studied longitudinally during active and subsequent inactive stage of their disease. During a 6 month period of study, we selected 20 consecutive patients with SLE who presented with a SLE flare. All patients fulfilled the 1982 revised criteria of the American College of Rheumatology for the classification of SLE. When patients entered the study, we performed psychiatric (CIS, RDC, STAI, HD, BDI, GHQ and MMS) psychosocial (GAS and VAS-P) scores assessment. One year later, we repeated the psychiatric and psychosocial assessment when patients showed inactive disease. The 20 patients evaluated were women, with a mean age of 34 y (SE 14.4, range 20 57). According to CIS evaluation, we diagnosed 8 (40%) psychiatric cases in the acute episode of SLE. The RDC diagnosis showed generalized anxiety in 5 patients, panic disorders in 2 patients and generalized anxiety plus depressive symptoms in one patient. One year later, when patients did not show disease activity, we diagnosed 2 (10%) psychiatric cases (P<0.05). When SLE patients were clinically inactive, they showed lower levels of psychological distress (GHQ scale, 1.8 vs 5.6, P<0.001), with a lower grade of anxiety measured by both HA (3.2 vs 8.2, P<0.01) and STAI-S (7.95 vs 20.90, P<0.001) scales. We also found a lower score in pain perception (VAS-P) (2.80 vs 4.25, P<0. 01) and higher occupational activity (VAS-P) (83.9 vs 66.2, P<0.01) and general functioning (GAS) (93.75 vs 83.50, P<0.05) during the inactive stage. No significant differences were found when we compared cognitive impairment, grade of depression and physical disability between inactive and active stages. We conclude that in SLE patients, psychiatric and psychosocial disorders during acute episodes are usually mild and seem to be related to the psychological impact of disease activity on patients. This type of psychiatric pathology is similar to that which would be expected in other groups coping with a stressful event, indicating that our patients did not react in a way specifically determined by their systemic disease. PMID- 11035433 TI - Elevation of proinflammatory cytokine (IL-18, IL-17, IL-12) and Th2 cytokine (IL 4) concentrations in patients with systemic lupus erythematosus. AB - Previous studies have indicated that the autoimmune phenomenon might be caused by an imbalance of T helper cell (Th) cytokines. We measured the plasma concentrations of three novel proinflammatory cytokines interleukin (IL)-17, IL 18, IL-12 and a key Th2 cytokine IL-4 in patients with systemic lupus erythematosus (SLE) and correlated the ratio of proinflammatory/Th2 cytokines with SLE disease activity index (SLEDAI). Plasma IL-12, IL-17, IL-18 and IL-4 concentrations of 36 SLE patients and 18 sex- and age-matched control subjects were measured by enzyme linked immunosorbent assay. All were significantly higher in SLE patients than control subjects (IL-12, mean+/-s.d. of 166.7+/-84.5 vs 93.5+/-39.2 pg/ml, P<0.001; IL-17, 76.5+/-45.7 vs 37.6+/-35.3 pg/ml, P=0.002; IL 18, 368.7+/-199. 5 vs 141.1+/-47.1 pg/ml, P<0.001; and IL-4, 27.1+/-15.3 vs 17.3+/-7. 2 pg/ml, P<0.05), and IL-18/IL-4 ratio correlated positively and significantly with SLEDAI score (r=0.435, P=0.006). We propose that SLE is characterized by an elevation of both Th1 and Th2 cytokines: the elevation of proinflammatory cytokine IL-12, IL-17 and IL-18 may trigger the inflammatory process in SLE and the elevation of IL-18/IL-4 ratio suggests an imbalance of cytokine profile to mediate the inflammatory response. PMID- 11035434 TI - High prevalence of antiphospholipid antibodies in leprosy: evaluation of antigen reactivity. AB - Antiphospholipid antibodies (aPL) have been reported not only in autoimmune disorders but also in various infectious diseases. Accumulating evidence indicates that beta2 glycoprotein I (beta2GPI) and prothrombin are the main proteins to which autoimmune aPL bind. The aim of this study was to evaluate the prevalence of different aPL in patients with leprosy. We included 51 outpatients (42 lepromatous and 9 borderline leprosy) without any clinical feature of the antiphospholipid syndrome (APS). 35 had lupus anticoagulant and 31 had anticardiolipin antibodies (aCL). Anti-beta2GPI antibodies were highly positive in 29/51 and anti- prothrombin antibodies (anti-II) were detected in 23/51. Almost all aCL and anti-beta2GPI were of IgM isotype, while IgG isotype was more frequent among anti-II. No statistical difference was found when aPL were evaluated in patients grouped according to their bacteriological status. Furthermore, patients under treatment (n=33) had a similar frequency of positive aPL compared to patients in vigilance (n=14). Assessing the specificity of antibody binding to CL and beta2GPI in ELISA by means of inhibition studies with cardiolipin-beta2GPI liposomes, leprosy and APS sera showed a similar behaviour. Comparable results were also found in both groups of patients when inhibition experiments with lysate of Mycobacterium leprae were carried out. In summary, leprosy-related aPL resemble those found in patients with APS but the immunoglobulin isotype is different, with IgM much more prevalent in leprosy patients. PMID- 11035435 TI - Lupus patients in an emergency unit. Causes of consultation, hospitalization and outcome. A cohort study. AB - The objectives were to determine causes of consultation, hospitalization and outcome in a cohort of lupus patients in an emergency unit. Patients with systemic lupus erythematosus (SLE) who visited the emergency department for consultation from 1 September 1996 to 17 May 1997 were included in the study. They were evaluated during the visit by looking at 100 variables such as demographic, socioeconomic, clinical, therapeutical, behavioral, (compliance), emotional (Beck depression inventory), disease activity, (Mex-SLEDAI), disease severity (Lupus SDI), chronic damage (SLICC-ACR), and physician's and patient's global assessments of severity. All causes of consultation, hospitalization and outcome were registered. Descriptive statistics, univariate analysis and multiple logistic regression were used for analysis. Significance was set at the 0.05 level. 180 patients were included. 164 were female, mean age 31.7/11.39 y, mean Mex SLEDAI score 3.8, mean SLICC-ACR 1.3. Fever, poliarthralgia and abdominal pain were the main causes of consultation with 26, 25 and 18 cases each. 49 patients were hospitalized and these were statistically different than non hospitalized patients in level of formal education (10.2 vs 11.8, P=0.03); compliance (7.6 vs 9, P=0.0001); malar rash (57% vs 82%, OR, 95% CI=0.28, 0.13 0.62, P=0.0008), chloroquine daily dose intake (45 vs 77 mg, P=0.04); disease severity in physician's global assessments (5.6 vs 2.1, P=0.0001) and Beck depression inventory (21 vs 16, P=0.01). Multiple logistic regression identified physician's global assessment, fewer ACR criteria and higher SLICC-ACR scores as the main variables associated with hospitalization. Five patients died; two with community acquired pneumonia, one with pancreatitis, multiple thromboses, and sepsis, one with pulmonary hemorrhage; and one with pulmonary thromboembolism. In conclusion, poor compliance, low level of formal education, severity, depression, lower ACR criteria and higher SLICC-ACR scores were important variables identified with hospitalization. Chloroquine use seemed to have a protective effect. Causes of death were related to infections and antiphospholipid syndrome. PMID- 11035436 TI - Antibodies to human myeloperoxidase in glomerular immune deposits of systemic lupus erythematosus. AB - Antibodies to human myeloperoxidase and cathepsin G have been detected in the serum of some patients with systemic lupus erythematosus. Therefore, the presence of antibodies to human myeloperoxidase and cathepsin G was examined in glomerular immune deposits. Glomerular basement membrane fragments were prepared from renal tissues obtained at autopsy from 19 patients with systemic lupus erythematosus. IgG was extracted from the glomerular basement membrane fragments and tested with sensitive immunoassays for antibodies to myeloperoxidase and cathepsin G. Antibodies to cathepsin G were not detected in the extracts but antibodies to human myeloperoxidase were found in extracts of one specimen. In the extract with 6M guanidine hydrochloride these antibodies were enriched 103-fold, compared to the initial supernatant of glomeruli, which served as a serum surrogate. The recovered antibodies to myeloperoxidase accounted for 12% of the recovered IgG. These findings add autoantibodies to human myeloperoxidase to the list of antibodies that have been shown to be present in glomerular immune deposits of patients with lupus glomerulonephritis. PMID- 11035437 TI - Increased serum soluble CD14, ICAM-1 and E-selectin correlate with disease activity and prognosis in systemic lupus erythematosus. AB - To improve monitoring of immunological and disease activity, we determined soluble markers of activity of the monocyte/macrophage system (sCD14) and the vascular endothelium (sE-selectin, sICAM-1) in patients with systemic lupus erythematosus (SLE) and primary Sjogren's syndrome (pSS) in comparison to patients with infections or sepsis. Concentrations of sCD14, sICAM-1 and sE selectin (soluble CD14, ICAM-1 and E-selectin, respectively) were measured in serum samples from patients with SLE and pSS, patients with sepsis, different infectious diseases and healthy controls using ELISA systems. Elevated levels of sE-selectin and sICAM-1 were detected in patients with SLE as well as sepsis, in contrast to patients with a localized infection (SLE and sepsis, respectively, versus infection P<0.001; Kruskal-Wallis test). Levels of sCD14 were persistently elevated in sera from patients with SLE, whereas these values decreased rapidly after effective therapy in patients with sepsis or infection. A continuous elevation of all of these three parameters was associated with a fatal outcome in patients with sepsis as well as in patients with SLE. Combined elevation of sCD14, sICAM-1 and sE-selectin correlates with the prognosis in patients with active SLE and indicates a remarkable immune activation involving the monocyte/macrophage system and the endothelium comparable to an activation found only in patients with sepsis. PMID- 11035438 TI - CD45 autoantibodies mediate neutralization of activated T cells from lupus patients through anergy or apoptosis. AB - The objectives were to provide estimates of the prevalence of autoantibody (Ab) directed to CD45 in lupus patients, identify the target autoantigen(s), and determine the ability of such reactivity to mediate neutralization of T lymphocytes. Sera from 64 patients were studied using 2 assays: Western blot and an ELISA with CD45 eluted from 3 cell lines as antigen (U937, Jurkat and Daudi). The role of carbohydrate specificity was investigated using enzyme digestion of blotted glycans, competition with sugars, and inhibition with lectins. Apoptosis was studied through annexin V binding, and cell cycle analysis using the propidium iodide method. AutoAb to CD45 were detected in 16/64 sera (25%) by Western blot, and 21/32 sera (66%) found positive in the ELISA. CD45 purified from Daudi cells was identified in the ELISA, but not in the blot. AutoAb were of the IgM and the IgG isotypes, but not specific for a particular cell type or CD45 isoform: 2 dominant specificities were recognized, one against p180, and another against high MW isoforms. Neuraminidase-induced enhancement of reactivity, together with the inhibitory effect of N-acetyl galactosamine and Dolichos diflorus lectin suggest that the epitopes are carbohydrates. AutoAb which were specific for activated CD4+T cells triggered the annexin V binding, and, in 2 of 4 cases, lymphocytes underwent apoptosis. In conclusion, carbohydrate conformational epitopes may be important as target antigens, and some CD45 autoAb have the capacity to neutralize activated T cells through anergy or apoptosis. PMID- 11035440 TI - Autoantibody responses to cardiolipin and DNA in infancy: association with lymphocytic panniculitis. AB - We report an infant with anticardiolipin (aCL) antibodies who presented with erythematous nodular skin lesions and elevated liver enzyme levels. The cutaneous manifestation was histologically lymphocytic lobular panniculitis with vasculitic and hemorrhagic changes. The infant also had low levels of anti-double stranded DNA (dsDNA) and anti-single stranded DNA (ssDNA) antibodies. There were no detectable antibodies to small nuclear ribonucleoproteins including Ro/SSA and La/SSB. His mother was consistently seronegative for any of these antibodies. Without corticosteroid therapy, cutaneous lesions resolved and anticardiolipin antibodies, but not anti-dsDNA and ssDNA antibodies, normalized within 16 months after the onset of the disease. Our patient demonstrated an uncommon presentation of aCL related cutaneous manifestation, the presentation with panniculitis being only the third such patient reported in the literature. Of great interest was the appearance of aCL antibodies and skin lesions during very early infancy. PMID- 11035439 TI - Systemic lupus erythematosus associated with moyamoya syndrome. AB - Moyamoya disease is an uncommon clinical entity, characterized by bilateral occlusion of the internal carotid artery and the development of collateral arteries. An 18-year-old Saudi male with systemic lupus erythematosus (SLE) presented with mild right hemiparesis, followed by recurrent ischemic stroke. Cerebral angiography showed bilateral internal carotid artery stenosis associated with the development of collateral circulation (moyamoya vessels). There was no evidence of active SLE or other risk factors for cerebral occlusion, such as antiphospholipid antibody syndrome. Medical and surgical interventions did not influence the poor outcome of the recurrent ischemic insults. PMID- 11035442 TI - Is hypoandrogenism a cause or a consequence of systemic lupus erythematosus in male patients? PMID- 11035441 TI - Erythema nodosum associated with antiphospholipid antibodies: a report of three cases. AB - Erythema nodosum is a dermatologic condition characterized by painful, erythematous nodules on the anterior surfaces of the lower extremities. Its association with a variety of conditions has been previously described. We present three cases of erythema nodosum in patients with elevated anticardiolipin antibodies. In one patient, a temporal relationship was seen in the simultaneous detection of antibodies and skin lesions. We propose an association between erythema nodosum and the antiphospholipid antibody syndrome (APS). PMID- 11035443 TI - General Summary. PMID- 11035444 TI - Association study on the DUSP6 gene, an affective disorder candidate gene on 12q23, performed by using fluorescence resonance energy transfer-based melting curve analysis on the LightCycler. AB - We introduced a new genotyping method, fluorescence resonance energy transfer based melting curve analysis on the LightCycler, for the analysis of the gene, DUSP6 (dual specificity MAP kinase phosphatase 6), in affective disorder patients. The DUSP6 gene is located on chromosome 12q22-23, which overlaps one of the reported bipolar disorder susceptibility loci. Because of its role in intracellular signalling pathways, the gene may be involved in the pathogenesis of affective disorders not only on the basis of its position but also of its function. We performed association analysis using a T>G polymorphism that gives rise to a missense mutation (Leu114Val). No evidence for a significant disease causing effect was found in Japanese unipolars (n = 132) and bipolars (n = 122), when compared with controls (n = 299). More importantly, this study demonstrates that melting curve analysis on the LightCycler is an accurate, rapid and robust method for discriminating genotypes from biallelic markers. This strategy has the potential for use in high throughput scanning for and genotyping of single nucleotide polymorphisms (SNPs). Molecular Psychiatry (2000) 5, 489-494. PMID- 11035445 TI - Forgetting hygiene. PMID- 11035446 TI - Physical activity and cause-specific mortality in the Whitehall study. AB - A prospective cohort study of London civil servants was used to examine the relation of physical activity to various causes of death. 6,702 men aged 40-64 y who participated in a baseline examination between 1969 and 1970 were followed up for 25 y during which time there were 2859 deaths. The association of two measures of physical activity (leisure time activity and usual walking pace) with cause-specific mortality was examined. Walking pace demonstrated inverse relations with mortality from all-causes, coronary heart disease (CHD), other cardiovascular disease (CVD), all cancers, respiratory disease, colorectal cancer and haematopoietic cancer following adjustment for risk factors which included age, employment grade, smoking, body mass index, and forced expiratory volume (P [trend]<0.05 for all). In analyses restricted to men without disease at entry, walking pace retained inverse associations with all-cause, CHD, other cardiovascular disease, and haematopoietic cancer mortality (P [trend]<0.05 for all). Leisure time activity was also inversely associated with mortality from all causes, CHD, other CVD, and all-cancers following adjustment for risk factors (P [trend]<0.05 for all). Eliminating deaths in the first 5 and 10 y of follow-up did not greatly alter these associations. It is concluded that physical activity may confer protection against death due to some cancers, in addition to reducing cardiovascular disease risk. PMID- 11035447 TI - Attendance at cultural events and physical exercise and health: a randomized controlled study. AB - The aim of this study was to assess the specific biomedico-social effects of participating in cultural events and gentle physical exercise effects apart from the general effect of participating in group activities. This was a randomized controlled investigation using a factorial design, where attending cultural events and taking easy physical exercise were tested simultaneously. The 21 participants, aged between 18 and 74 y were from a simple random sample of people registered as residents in Umea, a town in northern Sweden. Among the 1000 in the sample, 21 individuals (11 men, 10 women) were recruited into the experiment. Two out of the 21 subjects dropped out and were discounted from our analysis. Nine people were encouraged to engage in cultural activity for a two-month period. Diastolic blood pressure in eight of these nine was significantly reduced following the experiment. There were no marked changes observed in either systolic or diastolic blood pressure in those not required to engage in any form of extra-cultural activity. A decrease in the levels of both adrenocorticotropical hormone (ACTH) and s-prolactin was observed in culturally stimulated subjects, whereas the average baseline s-prolactin level of 7 ng/l for the non-culturally stimulated group was unchanged after the experiment. Physical exercise produced an increase in the high density lipoprotein (HDL) cholesterol level and in the ratio of HDL to LDL (low density lipoprotein). It was concluded that cultural stimulation may have specific effects on health related determinants. PMID- 11035448 TI - Grant funding of health voluntary organizations by Scottish health boards. AB - The objective of this study was to identify grant funding of voluntary organizations by Scottish health boards in the financial year 1997-1998. Scottish health board financial commitments to voluntary organizations were surveyed. Fifteen health board Directors of Finance participated in the study. The outcome measures were relative and absolute commitments of grant expenditure to the voluntary sector by health boards in Scotland. Total expenditure of Scotland's 15 health boards ranged from zero to 764,910 pounds sterling. One health board did not support any voluntary organizations while the greatest number supported by an individual health board was 43. Of health boards that made grants to voluntary bodies the range of expenditure per head of resident population was 0.09 pounds sterling-3.00p pounds sterling. The average grant expenditure to voluntary organizations ranged from 1,839 pounds sterling to 30,308 pounds sterling. The most substantial funding fell to voluntary bodies within the fields of mental health, alcohol and community elderly care. However, there was substantial variation between health boards in whether these bodies were funded, and to what extent funding was given. In conclusions, health boards have conflicting practices in funding voluntary organizations. Although there may be some variation in the needs of voluntary bodies across health boards, this is unlikely to explain the scale of the variation. Further work is required to explain this phenomenon and to propose policies for the support of the voluntary sector by the National Health Service that are acceptable to both funders and providers of services. PMID- 11035449 TI - Welfare benefits advice in primary care: evidence of improvements in health. AB - It is not uncommon for welfare benefits advice organisations to offer services in primary care settings. Given the link between deprivation and poor health, the maximising of individual income in this way may also be expected to improve health. However, such improvement has hitherto not been successfully measured. This paper reports on a small study of such a service, provided by the local Citizens Advice Bureau. Statistically significant increases in SF-36 scores were measured for those whose income increased as a result of receiving advice, despite the prevalence in the group (average age 56 y) of chronic disabling conditions such as arthritis and sensory impairment. These findings suggest that 'prescribing advice' is a health intervention which is appropriately situated in primary care. PMID- 11035450 TI - Small area variation in hospital admission: random or systematic? AB - For some conditions hospital admission is mandatory. This should lead to low variability in admission rates and no effect on admission rate of distance from hospital. If admission is discretionary, we would expect high variability in small area admission rates, and a decline in admission rate as travel time to hospital increases. We wanted to see if non-random variability of admission rates, as measured by the systematic coefficient of variation (SCV), and distance decay, as estimated in regression models, were related. We examined variability and travel time dependence of hospital admission for seven conditions in 62 small (mean population 9900) areas of Surrey, England. Age and sex standardized admission ratios (SAR) were calculated, and their dependence on travel time, adjusting for deprivation, were estimated by linear multiple regression adjusted for spatial correlation. Deprivation was measured by Jarman's score, and time by computerized estimates of drive time to the nearest acute hospital. We found an inverse relationship between time to hospital and admission ratio for ischaemic heart disease, bronchopneumonia and chronic bronchitis. Admission ratios for diabetes mellitus and stroke were related to neither deprivation nor time. For these seven conditions there was no simple relationship between SCV and travel time dependence. PMID- 11035451 TI - Effects of handicap on life expectancy: the case of China. AB - The purpose of this study was to quantify and partition the expected years of life with and without handicap for the Chinese population according to various types of handicaps, age-sex groups and regions. A large-scale sample survey on handicapped persons conducted in 1987, and the 1990 population census constitute the basis for computing the expected years of life free of handicapped condition using the method proposed by Sullivan. The expected years of life with handicap for the Chinese population in childhood (0-14 y), working ages (15-64 y) and the elderly (65 y+) were 0.40, 1.78, and 3.44 for males and 0.34, 1.69, and 4.55 for females. For the Chinese males over 65 y of age, there were about 1.83 expected years of life with aural handicap and 0.59 expected years of life with ocular handicap. For the Chinese females over 65 y of age, there were about 1.87 expected years of life with aural handicap and 1.16 expected years of life with ocular handicap. The burden of living with handicap is greater for females and the elderly. This general pattern hold for all types of handicap except for skeletal handicap. The expected years of life with handicap for the Chinese population provide useful information for setting public health policies, despite the difficulty in making comparisons with the similar data in other countries. PMID- 11035452 TI - Cancer survival rates and GDP expenditure on health: a comparison of England and Wales and the USA, Denmark, Netherlands, Finland, France, Germany, Italy, Spain and Switzerland in the 1990s. AB - Health funding is central to public health planning and clinical practice, hence this comparison of GDP health expenditure and five year post-diagnostic cancer survival rates of England and Wales with the USA and eight European countries. The three lowest proportional GDP health expenditures over the period 1980-1990 were Denmark, England and Wales, and Spain. The USA had the highest proportional GDP expenditure, followed by France, Germany, and The Netherlands. Overall the USA had the best cancer survival rates in the 14 sites reviewed, followed by Switzerland, The Netherlands, and Germany. The least successful were Spain, England and Wales, and Italy. In respect to the high incidence cancers, colorectal, lung, and female breast cancers, England and Wales survival rates were the poorest of all ten countries, followed by Denmark and Spain. Higher GDP health expenditure and longer survival rates for each gender were significantly correlated indicating a possible association between fiscal input and clinical outcomes, which poses problems for the development of effective public health. PMID- 11035453 TI - A comparison of normative and subjective assessment of the child prevalence of developmental defects of enamel amongst 12-year-olds living in the North West Region, UK. AB - Analysis of data from a 1996-97 cross sectional epidemiological study of the dental health of a sample of 12-y-old children living in Crewe, in north west England was used to compare normative and subjective assessment of developmental defects of enamel. Five hundred and twenty two 12-y-old children from secondary schools in Crewe were examined. One hundred and eighty two children (34.8%), had home post codes within the optimally fluoridated part of Crewe. Using the Developmental Defects of Enamel Index, 178 children (34%) in Crewe were normatively identified as having enamel defects present on their upper incisors. Thirty five children (6.7%), were unhappy with the appearance of their upper incisors because of marks that would not brush off. Neither the normative nor the subjective assessment of enamel defects demonstrated any difference in prevalence between the fluoridated and non-fluoridated areas. In Crewe, one in 20 children normatively diagnosed as being free from enamel defects were unhappy with the appearance of their upper incisors because of marks that would not brush off. There are differences in perception between dental professionals and 12-y-old children over the presence and relevance of developmental defects of enamel. Further research is required if we are to understand the difference in professional and lay perceptions of developmental defects affecting upper incisor teeth. PMID- 11035454 TI - Do patients know which healthcare professionals are doctors? AB - Using an anonymous questionnaire survey this study aimed to determine patients' knowledge of professional qualifications and which healthcare professionals are required to have undergone medical school training. Four hundred patients were questioned equally distributed between four general practices in Roehampton, London. The mean age of the sample was 43 y. Gender ratio was skewed with 31% men and 63% women (6% missing data). Amongst the three professional groups, just over 50% scored correctly. Almost 25% of respondents failed to correctly identify the listed doctors and medical non-doctors. The mean score of incorrectly identified alternative practitioners was 13%. There were significant age-related differences for scores, identified by chi-square and trend analysis, with the younger groups scoring better. Patients were largely unsure in identifying professional qualifications. It is concluded that confusion exists amongst patients as to which health-workers are doctors. Listing of professional qualifications does not help in clarifying this. Health-workers whose work involves invasive procedures or drug therapy are more likely to be thought of as medically qualified doctors. This confusion appears not to exist with alternative practitioners. Given the growing regard to appropriate and useful skill mix of health professionals (patients are increasingly seen in the first instance by non-doctors, eg nurse practitioners), it would seem appropriate that patients are made fully aware of who they are consulting. PMID- 11035455 TI - Cigarette smoking: knowledge, attitudes and behaviour in an adult population in Italy. AB - The study explores knowledge, attitudes, and behaviour regarding cigarette smoking and related factors in an adult population. A total of 935 parents of children attending the eighth class of ten randomly selected primary schools in Catanzaro (Italy) received a questionnaire consisting of questions on demographic and socioeconomic characteristics, knowledge, behaviour and attitudes about cigarette smoking. Six hundred and sixty-nine parents returned the questionnaire, with a response rate of 71.5%. Knowledge of risk associated to smoking was significantly higher in more educated subjects and in past smokers compared to current. Current and past smokers were respectively 39.6% and 17.2%, and current smokers were younger, not married, less prone to consider smoking as a major risk for their health and more likely to live with other smokers compared to past and never smokers. Females of higher education were more likely to be current smokers, whereas male current smokers were more likely to be less educated compared to past or never smokers. The results strongly recommend the provision of accurate information about the health consequences related to smoking, with a more intensive involvement of health care providers, particularly targeted to women and younger age groups. PMID- 11035456 TI - Improving the quality of spirometry in an epidemiological study: The Renfrew Paisley (Midspan) family study. AB - Population studies in Britain and elsewhere report deficiencies in quality of pulmonary function measurements. Methods were tested to improve the standardisation of spirometry in an epidemiological study. The spirometer provided visual feedback about acceptability and reproducibility to American Thoracic Society (ATS) standards. After 14 weeks technicians (research nurses) were given feedback and further training. Measurements were repeated in a 5% sample. Participant characteristics and technical factors (technician and technician feedback) predicted unacceptable forced expiratory volume in 1 second (FEV1) and excessively variable FEV1 and forced vital capacity (FVC). Only participant characteristics predicted unacceptable FVC. Feedback to technicians reduced test failure for FEV1 by half and excessive within-session variability by one-third. In the reproducibility study, coefficients of variation for FEV1 and FVC were 3%. Epidemiological studies can achieve standards of between-session reproducibility for spirometry comparable to levels reported by pulmonary function laboratories. Performance feedback to technicians improves the level of minimally acceptable spirometry, and within-session reproducibility. PMID- 11035457 TI - Hypertension awareness, detection and treatment in a university community: results of a worksite screening. AB - This study aimed to assess knowledge levels, explore the extent of undiagnosed hypertension, record previous screening activity and monitor the outcomes of previous hypertension treatment. To this end, information was collected from volunteer members of a university community, by means of a self-report questionnaire, about their personal characteristics, knowledge and experience of hypertension and stroke, and previous blood pressure measurements. In addition, their current blood pressure was recorded. A total of 653 staff and students completed the questionnaire and had their blood pressure measured. Analysis revealed that 82% were normotensive (diastolic blood pressure <90 mmHg). Hypertension was significantly associated with age, self-reported excess weight P<0.001) and marginally with self-reported non-healthy eating (P=0.06). Of the volunteers, 57% could not provide an accurate definition of a stroke. Knowledge levels were significantly and positively related to experience of stroke, healthy eating, not smoking and a recent blood pressure check. Of the respondents, 30% stated that they had not had their blood pressure measured in the previous two years. 51% of known hypertensives were not controlled. 68% of volunteers with diastolic blood pressure >89 mmHg were previously unaware of a potential hypertension problem. Hypertension rates amongst the university volunteers are higher than those recently recorded from a population sample. Scope exists for increasing knowledge and awareness, and for raising both screening rates and treatment outcomes. Improvements in these areas are required if current public health targets for heart disease and stroke are to be achieved. Worksite screening programmes can contribute to this endeavour. PMID- 11035458 TI - Consumption of prescribed and over-the-counter medicines with prolonged oral clearance used by the elderly in the Northern Region of England, with special regard to generic prescribing, dose form and sugars content. AB - As the longevity and population of elderly people has increased, the use of regular long-term medication for chronic medical problems has become more common. Medicines with prolonged oral clearance, for example syrups and chewable tablets, are commonly used in the elderly, many of whom retain their natural teeth into old age. These medicines may threaten dental health if they contain acidogenic sugars and are used long-term. As a part of an overall study of medication use in the elderly, three surveys were undertaken to assess the numbers of prescriptions and quantities of prescribed and 'over-the-counter' medicines with prolonged oral clearance dispensed during a 1 y period (1994), nationally and regionally. Of the 0.51 million litres of liquid oral medicines dispensed potentially for regular and long-term use by the elderly in the Northern Region in 1994, 94% was prescribed in primary care and 4% was sold over-the-counter from community pharmacies. When the effect of generic prescribing upon the sugars content of these medicines was considered, 96% of the volume of proprietary liquid oral medicines dispensed in primary care was sugars-free compared with 9% of generic liquid oral medicines. Of the 0.1 million litres of 'over-the-counter' liquid oral medicines sold in the Northern Region during 1994, 49% were sugars-free. In conclusion, although many prolonged oral clearance medicine preparations are sugars-free, due to generic prescribing a large proportion of the quantities dispensed for possible long-term use in the elderly are sugar-containing liquid oral medicines. In view of the increasing numbers of dentate elderly who require long-term medication, this is of some concern. The role of health professionals in raising awareness of the impact of generic prescribing on the sugars content of medicines is crucial if consumers are to benefit from the sugars-free option. PMID- 11035459 TI - Are Armed Forces infants more at risk than civilian infants? AB - The aim of the present study is to compare the health status of Armed Forces and civilian infants, accounting for social class. In a prospective cohort study, demographic data were obtained from mothers of liveborn infants from 436 civilian and 162 Armed Forces families. Birth details were taken from hospital maternity and child health systems. A six month follow-up was completed by health visitors. Standard social class classification, based on occupation, was used for civilian families and a new equivalent scheme for military personnel. No significant differences were found between civilian and military infants for birthweight, prematurity and failure to thrive. Military infants had significantly more hospital admissions (P=0.015) and accident and emergency attendances (P=0.002) mainly accounted for by the 'manual' social classes of the Armed Forces. Infant health status of civilian and military babies did not differ overall. Increased uptake of hospital services by military families can be explained by local circumstances. PMID- 11035460 TI - Child abuse among working children in rural Bangladesh: prevalence and determinants. AB - The paper aims to improve our understanding about the prevalence and determinants of child abuse in rural Bangladesh. Data from the 1995 sample survey of 4643 children aged 10-15 y in 150 villages were used. Findings revealed that 21% of the children were in the labour force although the Bangladeshi laws prohibited child labour. The prevalence of child abuse and exploitation was widespread in Bangladeshi villages as 2.3% of all children were physically abused, 2% were financially exploited, 1.7% were forced to involve in inappropriate activities, and 3% were forced to work for long hours. The prevalence of physical assault was much higher among younger children although the probability of other types of abuse was higher among older children. Boys were more exposed than girls to abuse of any kind. Poverty was also significantly associated with child abuse. Multivariate analysis suggested that the out-of-school children and the children of illiterate, landless and unskilled labourers were more likely to be abused than others when age and sex of children were controlled. The paper concludes that raising public awareness against child abuse and promoting preventive measures should be adopted to reduce child abuse in Bangladesh. PMID- 11035461 TI - Day care centres as an institution for health promotion among needy children: an analytical study in Sao Paulo, Brazil. AB - OBJECTIVE: To evaluate the role of day care centres in the nutritional state of children belonging to a low-income population, comparing the nutritional state of the children in the day care centres with children in the same population who were given other types of day care. DESIGN: Analytical cross-sectional study. SETTING: Public day care centres/primary health care. PARTICIPANTS: Children aged 0-6 yr attending in 4 public day care centres (n=446) and sample of population obtained during vaccination campaign (n=1626). MEASUREMENT: Nutritional evaluation was conducted through weight and height measurements, using as a standard the NCHS (USA) table and Gomez and Waterlow methodology. The type of daily care received by the children was assessed through a questionnaire answered by the responsible persons during the vaccination campaign. RESULTS: The nutritional status of children attended in the day care centers was better than children of the same community receiving other types of daily care (OR=0.48; CI 95%=0.36-0.65;P>0.0001). The improvement was related to more than 1 year of enrollment in the day care (OR=0.74; CI 95%=0.57-0.96;P=0.02). PMID- 11035462 TI - Sexual life style, drug habit and socio-demographic status of drug addicts in Bangladesh. AB - The sexual life style, drug habit and socio-demographic status of 253 male hospitalized drug addicts were investigated. This study was conducted during the period June 1997 to July 1998, and each case was selected by the random sampling method. The research instrument was an interviewer-administered questionnaire, the sexual habits, number and quality of sex partners, use of condoms, sexually transmitted diseases, etc., were considered as indicators of the sexual life style of the drug addicts. Eighty-eight percent (n=233) of the addicts were heterosexual. Bisexuality was found only in 7% (n=18) of the addicts. Eighty seven percent (n=240) of the addicts have multiple sex partners of either commercial or residential category. Most of the drug addicts (72%,n=181) did not use condoms. Fifty-seven percent (n=145) of the addicts were observed to have sexual diseases. As indicators of a drug habit, starting drug, choice of drug, period of addiction, sharing of needles, etc., were included. Cannabis was the starting substance for 59% (n=149) of the addicts. Heroin was the drug of choice for 60% (n=157) addicts. A single drug was taken only by 8% (n=20) of the addicts and the remaining 92% (n=233) took multiple drugs. The drug addicts (n=97) who used mostly injection (87%,n=84) shared needles. Education, occupation, income, age, marital status, influencing factors for addiction were considered as socio demographic characteristics. Young adults (79%,n=199), secondary educated (46%,n=116), low-mid income (60%,n=150), businessmen (46%,n=150) and married (60%,n=151) people were found highly involved in addiction. Self-curiosity and a friend's incitement were revealed as the most important influencing factors for taking drugs. PMID- 11035463 TI - Drug utilization and nutrition patterns among children from indigent and emigrant families in Crete, Greece. AB - AIM: To examine pharmaceutical needs, prescribed drugs, knowledge about pharmacotherapy and dietary patterns among indigent children. PATIENTS: 101 children (< or =18 y old), typically poor, from low-income families and emigrants coming back home from other countries, registered under Social Care in Chania, Crete. The control group comprised 81 Social Security insured children (< or =18 y old). The length of the study was from January 1995 to December 1997. RESULTS: The socio-demographic profile of the Social Care children reveals a no schooling rate of 9.7% vs 0% of the control group. The most common disease diagnosed in the indigent children was bronchitis (18.5%) compared with respiratory infection (14.6%) in the control group. Tuberculosis was diagnosed in 2.0% of the Social Care indigent children and in none of the insured children. The most frequently prescribed drug category in both groups as Defined Daily Doses (D.D.D.) was for the respiratory system (32.4% vs 21.2%), while antibiotics were the most expensive (41.6% in the indigent vs 54.9% in the control group). Only 27.4% of Social Care indigent patients, versus 51.2% of insured patients, understood the instructions regarding the proper use of their drugs. Regarding the dietary patterns, significant differences were found in the consumption of breakfast every day (73.7% vs 87.7%), red meat > or =4 per week (0% vs 6.3%) and fruits often (60.1% vs 75.0%). CONCLUSION: The present study emphasises the need for more information on drug use and the necessity for a continuing health educational intervention among indigent children. PMID- 11035464 TI - Commissioning regional specialist services-HIV and AIDS. Experiences of a consortium arrangement in the North East of England. AB - The introduction of effective double combination therapy for HIV disease in 1996 and triple therapy in 1997 had cost implications for both purchasers and providers of regional specialist services. There is evidence that more patients presented for treatment. A consortium purchasing approach was undertaken in the North East of England to manage the introduction of combination therapy, to ensure that therapy was available for those who could benefit and to minimise the financial risk for individual health districts. The strengths and weaknesses of such an approach are discussed as well as the potential application to commissioning other specialist services. PMID- 11035465 TI - Control of onchocerciasis in Nzerem-Ikpem, Nigeria: baseline prevalence and mass distribution of ivermectin. AB - Data from a survey of endemicity of human onchocerciasis and subsequent mass distribution of ivermectin to control this disease in 9 villages of the Nzerem Ikpem community in the Imo River basin of Nigeria are presented. Of the 1126 persons examined, 46.2% were positive for skin microfilarial, 29.2% for leopard skin, 34.2% for palpable subcutaneous nodules and 14.8% for various types of onchodermatitis. Ivermectin was distributed to a total of 1934 persons living in 233 households. The treatment coverage rate was 51%. 12.8% of those treated reported various mild reactions to the microfilaricide. Implications for community participation and long term sustenance of mass distribution programs are discussed. PMID- 11035466 TI - How do rural households perceive and prioritise malaria and mosquito nets? A study in five communities of Nigeria. AB - The aim of this study was to determine households' levels of prioritization and perception of malaria, ordinary mosquito nets and insecticide-treated nets (ITNs). A cross-sectional survey was conducted in five malaria holo-endemic communities in Enugu State, South-eastern Nigeria. The household heads or the representatives from randomly selected households were interviewed, using a pre tested interviewer-administered questionnaire. The majority of the respondents had a good knowledge about malaria and the use of ordinary mosquito nets to prevent malaria. However, few knew about the existence of ITNs. Most respondents also stated that malaria was a priority problem and perceived some risk of contracting it. Despite the high level of knowledge about the use of mosquito nets, only 14.0%, 15.7%, 9.6% and 8.0% of the respondents from four of the communities had ever purchased any type of mosquito nets, except in Orba where the proportion was 50.3%. However, more than 80% in all the communities expressed a desire to buy insecticide-treated mosquito nets for the prevention of mosquito bites. There was considerable knowledge about malaria and the use of mosquito nets to prevent it. There were also high levels of prioritization of the disease, mosquito nets and ITNs which signalled the possibility of establishing sustainable community-based ITN programmes, especially as households wanted to buy the ITNs. PMID- 11035467 TI - Horizontal transmission of HBV infection among students in Turkey. AB - In this study, the HBsAg carrier state and the role of horizontal transmission were investigated among primary and high school students in southeastern Anatolia where HBsAg seropositivity is remarkably high. In total, 350 students from primary school first grade, 350 students from fifth grade, 400 students from high school eleventh grade and 400 healthy adults as a control group were studied. In all cases HBsAg and anti-HBs were screened by ELISA. HBsAg positivity was 2.4% in first grade, 6.1% in fifth and 6.7% in eleventh grade students. Anti-HBs positivity was 14% in first grade, 20% in fifth and 21% in eleventh grade students. HBsAg positivity was 9% and anti-HBs, 49% in the control group. There is a significant difference between first and fifth grade students for HBsAg positivity (2.1% vs 6.1% and P<0.05). This difference decreased during the high school years (6.2% and P>0.05). There is also a similar statistically significant difference for anti-HBs positivity during the primary school years (14% vs 20%, P<0.05). These findings show that the risk of horizontal transmission of HBV is especially important during elementary school years between the ages of 7 and 11 y. All infants or at least elementary school first grade students in Turkey should have HBV vaccinations. PMID- 11035468 TI - Geographical display of health information: study of hepatitis C infection in Karachi, Pakistan. AB - The prevalence of hepatitis C and other infections is increasing in urban areas of developing countries. Data on such diseases are often limited to facility based information. However, even this is not available in a usable form to health care providers, health managers and policy makers. We present a simple technique for visually displaying facility based prevalence information on hepatitis C using basic geographic information system (GIS) techniques. We display the prevalence of hepatitis C for the city of Karachi, Pakistan for the first time. The distribution tends to indicate that there are areas of higher prevalence located in specific districts. There is also a trend of higher prevalence in less affluent urban areas. Such simple applications of mapping technology are useful for rapidly summarizing and displaying information in a contextually and spatially meaningful fashion, and its use should be encouraged for displaying health indicators in developing countries. PMID- 11035469 TI - Unintentional injury mortality and socio-economic development among 15-44-year olds: in a health transition perspective. AB - Injury imposes one of the greatest health risks in terms of mortality and morbidity among 15-44-y-olds. There is evidence that socio-economic development (SED) is related to injury risk, but the findings are inconsistent. We aimed to study the magnitude, pattern and relative importance of unintentional injury mortality (UIM) in relation to SED in this age group. Cross-sectional data on UIM by age-sex specific groups were obtained for 54 countries from the World Health Statistics Annuals 1993-1995. The relationship between UIM and SED (measured in gross national product (GNP) per capita) was studied using two methods: (1) with regression analysis, and (2) by categorizing the data into four income-based country groups and then comparing the differences in their mean values. The results were: (1) UIM rates were inversely correlated with GNP per capita and the relationship became stronger with increasing age (r=-0.22 for both sexes in the 15-24-y-olds, r=-0.65 for males, r=-0.54 for females in the 35-44-y-olds); (2) there was an increase in UIM rates between low-income and lower-middle-income countries (LoMIC), but a decrease between LoMIC and upper middle-income (UpMIC), and finally also a significant decrease between UpMIC and high-income countries in most age-sex groups (ie P<0.005 for males, P<0.05 for females in the 35-44-y olds). The highest rates of UIM were in LoMIC for all age-sex groups. Male rates were consistently higher than female in all age groups. In conclusion, SED was inversely related to UIM. There was an initial positive relation between GNP per capita and UIM, which became negative with increasing GNP per capita. We also found a health transition that had taken place in all country groups. PMID- 11035471 TI - Spinal cord injury and quality of life measures: a review of instrument psychometric quality. AB - INTRODUCTION: Quality of life (QOL) is often considered the primary endpoint in research, clinical medicine, and health promotion when impairments are incurable or insufficiently understood. For spinal cord injured (SCI) persons extended life spans and the need for life-long follow-up make it important to expand the outcome parameters of medical care and health services to include QOL measures. STUDY DESIGN: Review. OBJECTIVES: The aim of this study was to evaluate QOL research in SCI persons from the perspective of current criteria for instrument psychometric quality developed by Medical Outcomes Trust (MOT). METHODS: Relevant articles were extracted from the Medline, Cinahl, and PsycLit databases for approximately a recent 30-year period (1966 - 1999). The keyword 'spinal cord injuries' was cross-indexed with 'quality of life', 'personal satisfaction' and 'life satisfaction'. A total of 105 articles were identified and 46 met our inclusion criteria: (a) report of original research; (b) evaluation of QOL by self-report questionnaires or scales; and (c) publication in English. RESULTS: The quality of the research designs varied widely. Most of studies were cross sectional with limited study populations. The number of instruments or different combinations of instruments nearly equalled the number of studies conducted. Most questionnaires were condition-specific and only used by the developers. The variety of instruments and the diversity of their core content made results difficult to compare, and evaluations and conclusions unpredictable and sometimes contradictory. The low psychometric standard of many instruments used further aggravates the interpretation of results. CONCLUSION: To improve future research in special populations such as the SCI, the MOT criteria for review of QOL instruments should be further disseminated and applied to reach desirable consensus of a limited number of standardised generic and condition-specific QOL measures. PMID- 11035472 TI - Epidural electrical stimulation of posterior structures of the human lumbosacral cord: 3. Control Of spasticity. AB - OBJECTIVES: The purpose of this study was to evaluate the effect of spinal cord stimulation (SCS) on severe spasticity of the lower limbs in patients with traumatic spinal cord injury (SCI) under close scrutiny of the site and parameters of stimulation. MATERIALS AND METHODS: Eight SCI patients (four women, four men) were included in the study. Levels of spasticity before and during stimulation were compared according to a clinical rating scale and by surface electrode polyelectromyography (pEMG) during passive flexion and extension of the knee, supplemented by a pendulum test with the stimulating device switched either on or off over an appropriate period. RESULTS: Both the clinical and the experimental parameters clearly demonstrated that SCS, when correctly handled, is a highly effective approach to controlling spasticity in spinal cord injury subjects. The success of this type of treatment hinges on four factors: (1) the epidural electrode must be located over the upper lumbar cord segment (L1, L2, L3); (2) the train frequency of stimulation must be in the range of 50 - 100 Hz, the amplitude within 2 - 7 V and the stimulus width of 210 micross; (3) the stimulus parameters must be optimized by clinically assessing the effect of arbitrary combinations of the four contacts of the quadripolar electrode; and (4) amplitudes of stimulation must be adjusted to different body positions. CONCLUSIONS: Severe muscle hypertonia affecting the lower extremities of patients with chronic spinal cord injuries can be effectively suppressed via stimulation of the upper lumbar cord segment. PMID- 11035474 TI - Hyperbaric oxygen (HBO) therapy for acute traumatic cervical spinal cord injury. AB - STUDY DESIGN: A retrospective study of spinal cord injury (SCI) treated with and without hyperbaric oxygen (HBO) therapy. OBJECTIVES: To report on the use of HBO in spinal cord injury. SETTING: Neurosurgical Unit, Tokyo, Japan. METHODS: Thirty four cases of hyperextension spinal cord injury without bone damage and previous history of surgical intervention were divided into two groups, with (HBO) or without (non-HBO) therapy. The neurological findings at admission and their outcomes were evaluated by means of Neurological Cervical Spine Scale (NCSS)1 and the average improvement rates in individual groups were compared. RESULTS: The improvement rate ranged from 100% to 27.3% with the mean value of 75. 2% in the HBO group, while these values were 100%, 25.0% and 65.1% respectively in the non HBO group. CONCLUSION: In the HBO group, the improvement rate indicated effectiveness in acute traumatic cervical spinal cord injury. PMID- 11035473 TI - A histopathological analysis of the human cervical spinal cord in patients with acute traumatic central cord syndrome. AB - STUDY DESIGN: We have applied conventional histochemical and morphometric techniques to study the changes within the human spinal 'hand' motor neuron pool after spinal cord injury in patients who presented with acute traumatic central cord syndrome (ATCCS). OBJECTIVE: To determine whether a reduction of large alpha motor neurons at the C7, C8 and T1 spinal cord levels underlies the mechanism which causes hand dysfunction seen in patients with (ATCCS). BACKGROUND: The etiology of upper extremity weakness in ATCCS is debated and injury and/or degeneration of motor neurons within the central gray matter of the cervical enlargement has been advanced as one potential etiology of hand weakness. METHODS: The spinal cords of five individuals with documented clinical evidence of ATCCS and three age-matched controls were obtained. The ATCCS spinal cords were divided into acute/sub-acute (two cases) and chronic (three cases) groups depending on the time to death after their injury; the chronic group was further subdivided according to the epicenter of injury. We counted the motor neurons using light microscopy in 10 randomly selected axial sections at the C7, C8 and T1 spinal cord levels for each group. We also analyzed the lateral and ventral corticospinal tracts (CST) in all groups for evidence of Wallerian degeneration and compared them to controls. RESULTS: A primary injury to the lateral CST was present in each case of ATCCS with evidence of Wallerian degeneration distal to the epicenter of injury. There was minimal Wallerian degeneration within the ventral corticospinal tracts. In the chronic low cervical injury group, there was a decrease in motor neurons supplying hand musculature relative to the other injury groups where the motor neurons sampled at the time of death were not reduced in number when compared to the control group. CONCLUSIONS: We hypothesize that hand dysfunction in ATCCS can be observed after spinal cord injury without any apparent loss in the number of motor neurons supplying the hand musculature as seen in our acute/sub-acute (n=2) and our chronic high injury (n=1) groups. The motor neuron loss seen in the chronic low level injury was felt to be secondary to the loss of C7, C8, and T1 neurons adjacent to the injury epicenter. PMID- 11035475 TI - Improvement of urological-management abilities in individuals with tetraplegia by reconstructive hand surgery. AB - OBJECTIVE: To evaluate whether reconstructive hand surgery could improve the ability of tetraplegic patients to perform clean intermittent self catheterization (CIC). MATERIALS AND METHODS: The subjects were patients with complete traumatic tetraplegia admitted to the Nagano Rehabilitation Center between 1977 and 1996. A total of 63 subjects were included in this study with an age range of 18-73 years (mean 38.2 years). Reconstructive hand surgery was performed on 44 hands (28 cases). Current urological conditions were assessed by interview or mail questionnaire. RESULTS: Fifty-one per cent (22/43) of the patients with C6 level of tetraplegia and 86% (12/14) of those with C7 or C8 neurological level of injury could perform CIC independently and only one subject needed assisted CIC. CONCLUSION: CIC is the preferred option for people with tetraplegia. Reconstructive hand surgery is thus recommended to make urological management more independent for a selected group of people with tetraplegia. PMID- 11035476 TI - Parathyroid hormone-related protein (1-34) and urothelial redifferentiation in the neuropathic urinary bladder. AB - STUDY DESIGN: A comparative study of immunostaining for parathyroid hormone related protein (1-34) (PTHrP (1-34)) in the vesical epithelium of biopsies obtained from patients with non-neuropathic bladder and those with neuropathic bladder. OBJECTIVES: To investigate the immunostaining for PTHrP (1-34) in the control cases and in neuropathic bladders showing (1) normal transitional epithelium, (2) hyperplastic transitional epithelium, and (3) squamous metaplasia. SETTING: Regional Spinal Injuries Centre, and Department of Cellular Pathology, Southport & Ormskirk Hospitals NHS Trust, Southport, Department of Pathology, Royal Liverpool University Hospital and the Departments of Clinical Chemistry and Cell Biology, The University of Liverpool, Liverpool, England. METHODS: Cold cup biopsies of bladder mucosa were taken from patients suffering from neuropathic urinary bladder when they were undergoing a therapeutic procedure in the urinary tract. Immunohistochemistry was performed on these biopsy specimens using a rabbit polyclonal antibody raised to a synthetic peptide corresponding to human PTHrP (1-34). Control group (n=10) consisted of archival biopsies taken from non-neuropathic bladders. RESULTS: In the control group, the transitional epithelium showed no immunostaining, or at the most, very faint positive staining was seen in the transitional epithelium of non-neuropathic bladder. Positive immunostaining to PTHrP (1-34) was seen in the normal transitional epithelium of neuropathic bladder in nine of 13 cases. Hyperplastic transitional epithelium showed positive immunostaining for PTHrP (1-34) in 11 of 13 biopsies from patients with neuropathic bladder. Immunostaining for PTHrP (1 34) was observed in the metaplastic squamous epithelium in 14 of 17 cases with neuropathic bladder. CONCLUSION: The transitional epithelium of non-neuropathic bladder showed no immunostaining, or at the most, very faint positive staining for PTHrP (1-34). In contrast to this, positive immunostaining for PTHrP (1-34) was observed more frequently in the vesical epithelium of neuropathic bladder. This observation opens up avenues for innovative therapy with PTHrP or its analogues for possible modulation of urothelial differentiation in the neuropathic bladder. PMID- 11035477 TI - Urologic status of 74 spinal cord injury patients from the 1976 Tangshan earthquake, and managed for over 20 years using the Crede maneuver. AB - OBJECTIVE: To evaluate the urologic safety of long-term Crede maneuver as bladder management in spinal cord injured patients. METHODS: Seventy-four paraplegics were included in this cross-sectional study. They were injured in the Tangshan earthquake in 1976. All patients have large volume (flaccid) bladders and have practiced the Crede maneuver for more than 20 years to expel urine. Current residual urine volume and urologic complications were investigated. RESULTS: 93.2% of patients have residual urine larger than 100 ml and 50% of cases larger than 300 ml. The prevalence of urologic complications is high: pyuria in 82.4%, urinary lithiasis in 31.3%, ureteral dilatation in 59.5%, hydronephrosis in 35.1% and renal damage in 16.2%. Men are more susceptible to upper urinary tract deterioration than women (P<0.05). CONCLUSION: The Crede maneuver is not safe for long-term use in spinal cord injury patients, especially in men. PMID- 11035478 TI - One year follow up of spinal cord injury patients using a reciprocating gait orthosis: preliminary report. AB - OBJECTIVE: To examine the influence of social, physical and psychological factors in determining the usage/non usage of reciprocating gait orthosis (RGO) in spinal cord injury (SCI) patients. DESIGN: Prospective clinical trial. SETTING: A large rehabilitation hospital in Rome, Italy. PARTICIPANTS: Twenty four SCI patients of traumatic aetiology (all fulfilling the criteria to prescribe the device). METHODS: Social, physical and neurological examination according to ASIA standards; psychological enquiry by means of the Eysenck Personality Questionnaire (EPQ) and the scale for self rating anxiety and depression of the Cognitive Behavioural Assessment. RESULTS: After 1 year follow up 11 (46%) of our patients no longer used the RGO. There was no statistically significant difference between patients who used the RGO and those who rejected the orthosis with regard to social and physical data. There was a significant difference (P=0.005 at the end of training and P=0.003 at 1 year follow up) with regard to functional ambulation level. With regard to psychological enquiry RGO-non users showed a higher frequency of values over the mean in the E scale (extroversion) of the EPQ than RGO-users (P=0.05). CONCLUSIONS: None of the identified parameters were useful to predict the use/rejection of the orthosis. Although they need to be confirmed, our psychological data suggest that extensive psychological testing could be useful to sharpen the ability to predict. PMID- 11035479 TI - Psychological wellbeing among carers of people with spinal cord injury: a preliminary investigation from South India. AB - STUDY DESIGN: A cross-sectional study that assessed people with spinal cord injury (SCI) and their carers who attended the 3-day health care program. OBJECTIVES: The study examined the nature and prevalence of the factors associated with psychological morbidity among carers of people with SCI. SETTING: A community reintegrated population of persons with SCI and their carers attended the 3-day program in the Department of Physical Medicine and Rehabilitation, Christian Medical College Hospitals, Vellore, South India. METHODS: Thirty-eight people with SCI and their carers participated in this cross-sectional study. RESULTS: Thirty (78.9%) carers of people with SCI were psychologically distressed. While carers were distressed, they were not significantly depressed. Educational level of carers and suicidal behavior of people with SCI were significantly associated with psychological distress. CONCLUSION: The findings of this study suggest that health care workers should have a high index of suspicion of psychological morbidity, in carers of people with SCI. Identification of risk factors may lead to useful target interventions. PMID- 11035480 TI - Long-term morbidity and mortality after spinal cord injury: 50 years of follow up. AB - OBJECTIVE: To determine the long-term mortality rate and the types of morbidity among all people with spinal cord injuries (SCI) that occurred during the 1948 Israel War of Independence. METHOD: Chart review and telephone interviews for collecting demographic data, injury characteristics, marital status, physical activities, employment, morbidity and mortality. RESULTS: Twenty individuals with SCI (19 males, one female). There was no regular follow-up during the first 20 years post injury. The most frequent morbidities were genito-urinary, cardiovascular and decubiti. Ten (50%) had died during this overall follow-up interval. The average age at death was 60 years. The cause of death was cardiovascular in six, neoplastic disease in two, pneumonia in one, and one died from an unknown cause. CONCLUSIONS: The data analysis showed that those who died participated less in physical activity and fewer were employed as compared to the survivors. PMID- 11035481 TI - Post-traumatic syringomyelia following complete neurological recovery. AB - OBJECTIVE: To describe the later neurologic deterioration secondary to the appearance of a post-traumatic syringomyelic cavity, in a patient who, in the initial phase, had an incomplete spinal cord lesion (ASIA C), which improved to ASIA E. METHODS: A 52-year-old male patient who, at the age of 19 (1965), suffered a spinal cord injury. He presented with a fracture of the sixth and seventh cervical neurological segment at the time of the lesion, evolving to ASIA E. Nine years after the traumatism, he began to feel pain accompanied by a sensory and motor deficit. RESULTS: With the aid of myelography and MRI, the existence of a syringomyelic cavity was detected, which extended from the fourth to the seventh cervical segments. The patient was operated on, on various occasions, placing a syringo-subarachnoid shunt. The neurological status of the patient continued to deteriorate and, at present, he has a complete lesion below the fourth neurological cervical segment with a partially preserved sensitive area up to T1. CONCLUSION: The development of the syringomyelic cavity could be one of the causes of later neurologic deterioration in patients with traumatic spinal cord injury with neurological recovery 'ad integrum' in the initial phase of spinal cord injury. PMID- 11035482 TI - Rotation flaps in the treatment of ischial pressure sores--the bigger the better. AB - Pressure sores are a common problem in paraplegic patients or those otherwise immobilised for long periods of time. Their surgical management is predominantly undertaken by plastic surgeons but other surgeons will also treat them from time to time. We present a case of an ischial pressure sore treated by a general surgeon with a small buttock rotation flap that subsequently broke down with recurrence of the pressure sore. The subsequent surgical management under our care was compromised by his earlier surgery but nevertheless achieved a satisfactory outcome. The need to perform repeat surgery in the future must therefore be borne in mind when planning the initial operation, so that the maximum number of reconstructive options are preserved. PMID- 11035483 TI - Placenta of discordant twins: lack of change in histochemically detectable enzyme activities. AB - We localised three important enzymes histochemically in placental trophoblasts from women who gave birth to dichorionic discordant twins, in which the co-twin was affected by foetal growth restriction (FGR). The enzymes studied were adenosine diphosphate-degrading enzyme (ADP-degrading enzyme, plasma membrane enzyme), cytochrome c oxidase (mitochondrial enzyme), and glucose-6-phosphatase (endoplasmic reticular enzyme). We compared these enzyme activities and their distribution patterns among placentas of the smaller (FGR) co-twin, larger co twin, pre-eclamptic singleton with FGR, and normal singletons with birth weight of appropriate for their gestational ages. In FGR co-twin placentas, the intensity and localisation pattern of these three enzymes did not differ from those seen in the larger co-twin and normal singleton placentas. Decreased ADP degrading activity and cytochrome c oxidase negative mitochondria, which were characteristic features of pre-eclamptic trophoblasts, were not observed in FGR co-twin placentas. These observations indicated that, in the FGR co-twin, enzyme histochemically detectable trophoblastic cell dysfunction may be absent, or if present, less prominent, compared with pre-eclamptic FGR. We previously reported that placental trophoblasts from singleton idiopathic FGR also showed no reduction in these enzyme activities. In mechanism and pathophysiology, FGR in dichorionic discordant twins may be quite different from pre-eclamptic FGR, but somewhat resembles idiopathic FGR, though all three disorders lead to placental insufficiency, resulting in limited foetal growth. PMID- 11035484 TI - Infant zygosity can be assigned by parental report questionnaire data. AB - A parental report questionnaire posted to a population sample of 18-month-old twins correctly assigned zygosity in 95%of cases when validated against zygosity determined by identity of polymorphic DNA markers. The questionnaire was as accurate when readministered at 3 years of age, with 96% of children being assigned the same zygosity on both occasions. The results validate the use of parental report questionnaire data to determine zygosity in infancy. PMID- 11035485 TI - Zygosity diagnosis in young twins by parental report. AB - This study reports on zygosity determination in twins of childhood age. Parents responded to questionnaire items dealing with twin similarity in physical characteristics and frequency of mistaking one twin for another by parents, relatives and strangers. The accuracy of zygosity diagnosis was evaluated across twins aged 6, 8, and 10 and across parents. In addition, it was examined whether the use of multiple raters and the use of longitudinal data lead to an improvement of zygosity assignment. Complete data on zygosity questions and on genetic markers or blood profiles were available for 618 twin pairs at the age of 6 years. The method used was predictive discriminant analyses. Agreement between zygosity assigned by the replies to the questions and zygosity determined by DNA markers/blood typing was around 93%. The accuracy of assignment remained constant across age and parents. Analyses of data provided by both parents and collected over multiple ages did not result in better prediction of zygosity. Details on the discriminant function are provided. PMID- 11035486 TI - The comparative constitution of twinship: strategies and paradoxes. AB - In both traditional and modern societies, twinship, as an unusual mode of reproduction, involves difficulties for social systems in maintaining consistent classification systems. It is proposed that the most prevalent response to twinship involves various 'strategies of normalisation' to defuse and contain the potential disruption. This proposition is illustrated and analysed in relation to ethnographic maternal drawn mainly (but not exclusively) from African communities in the twentieth century. Following a discussion of twin infanticide as the most extreme of the normalising strategies, the article concludes by identifying a number of paradoxes in the social construction of twinship. PMID- 11035487 TI - Improved indices of insulin resistance and insulin secretion for use in genetic and population studies of type 2 diabetes mellitus. AB - Homeostasis model assessment (HOMA) provides indices of insulin secretion (beta) and insulin resistance (R) derived from fasting plasma glucose (FPG) and fasting plasma insulin (FPI) levels. However, these indices could not account for a significant heritability of fasting plasma glucose (FPG) (h2 = 0.75, P<0.01) in a group of 214 female twins. This result is consistent with a misclassification between effects due to insulin secretion and resistance in the HOMA indices. We report here evidence of such misclassification in the HOMA indices and describe a minor modification to the model which corrects it. Direct measures of insulin resistance (euglycaemic clamp) and secretion (i.v. glucose bolus) were obtained in 43 non-diabetic subjects. Heritability was estimated by statistical modelling of genetic and environmental influences in data from 214 non-diabetic female subjects. Modified HOMA (HOMA') indices were obtained from beta' = (Ln(FPI) - c)/FPG and R' = (Ln(FPI) - c)*FPG where c is a constant derived from regression analysis of Ln(FPI) vs FPG. Indices from both models correlated with the direct measures similarly (r = 0.63 (R), 0.49 (R'), 0.45 (beta), 0.39 (beta'), all P< 0.01). Directly measured insulin resistance and secretion were not significantly correlated (r = 0.13, P = 0.21). However, unmodified HOMA-beta and R were strongly related (r = 0.78, P<0.0001 vs. 0.13) demonstrating substantial misclassification. The relationship between beta' and R' (r = 0.13) was not different from that between the two direct measures and significant heritability of beta' (h2 = 0.68, P<0.01) and R' (h2 = 0.59, P<0.05) was evident in the twin data. The proposed modification to HOMA significantly reduces misclassification and reveals separate components of insulin resistance and insulin secretion in the heritability of FPG. PMID- 11035488 TI - The effect of apolipoprotein(a)-, apolipoprotein E-, and apolipoprotein A4- polymorphisms on quantitative lipoprotein(a) concentrations. AB - The effects of apolipoprotein (a), apolipoprotein-E, and apolipoprotein-A4 isoforms on quantitative lipoprotein(a) [Lp(a)] levels were assessed in a sample of 142 Dutch families consisting of two parents and their adolescent twin offspring. A total heritability of 95% was estimated for plasma Lp(a) concentrations. The largest part of this heritability was due to the apo(a) locus which explained 61% of the total variance in Lp(a) levels. The pattern of familial correlations for the residual part of the Lp(a) variance that could not be attributed to the apo(a) isoforms, suggested genetic influences on the residual variance. We addressed the question whether this residual genetic variance could be ascribed to the apoE or the apoA4 locus. A simultaneous analysis of all three loci showed that both the apoE and the apoA4 polymorphism did not contribute significantly to Lp(a) variation. PMID- 11035489 TI - Defining discordance in twin studies of risk and protective factors for late life disorders. AB - In studies that employ matched pair analysis to identify environmental exposures important for a disorder, criteria for discordant pairs are seldom discussed. Yet several assumptions concerning the definition of discordancy may have considerable influence over what results are found. Problems are exacerbated when age of onset for a disorder is late in life. We propose a new set of criteria for defining discordant pairs in studies of dementia, taking into account duration of discordance and competing causes of mortality, and evaluate the consequences of choosing alternative definitions of discordancy. PMID- 11035490 TI - Structured latent growth curves for twin data. AB - We describe methods to fit structured latent growth curves to data from MZ and DZ twins. The well-known Gompertz, logistic and exponential curves may be written as a function of three components - asymptote, initial value, and rate of change. These components are allowed to vary and covary within individuals in a structured latent growth model. Such models are highly economical, requiring a small number of parameters to describe covariation across many occasions of measurement. We extend these methods to analyse longitudinal data from MZ and DZ twins and focus on the estimation of genetic and environmental variation and covariation in each of the asymptote, initial and rate of growth factors. For illustration, the models are fitted to longitudinal Bayley Infant Mental Development Scale data published by McArdle (1986). In these data, all three components of growth appear strongly familial with the majority of variance associated with the shared environment; differences between the models were not great. Occasion-specific residual factors not associated with the curve components account for approximately 40% of variance of which a significant proportion is additive genetic. Though the growth curve model fit less well than some others, they make restrictive, falsifiable predictions about the mean, variance and twin covariance of other (not yet measured) occasions of measurement. PMID- 11035491 TI - Genetic epidemiology of complex traits. PMID- 11035492 TI - [Advertising in cadernos de Saude Publica] [In Process Citation] PMID- 11035493 TI - [The concept of space in epidemiology: a historical and epistemological interpretation]. AB - This study provides an interpretation of the concept of space in epidemiology. The authors highlight that the epistemological orientation of the space concept in epidemiology is the theory of disease, emphasizing the importance of the concept of specific etiologic agents and their transmission as the central structure for grasping the relationship between space and the body. Characterization of the space for circulation of etiologic agents was the epistemological base shaping the use of various theoretical developments in geography, allowing for the construction of different explanatory watersheds in the concept of space. The article specifically analyzes the Latin American watershed, reviewing the main authors orienting these studies, like Pavlovsky, Max Sorre, and Samuel Pessoa. The authors highlight Milton Santos' thinking as a fundamental reference in recent research on the social organization of space and disease emergence or prevalence. The authors also approach contemporary changes in the understanding of space as they are reflected in epidemiological studies. PMID- 11035494 TI - [Debate on the paper by dina czeresnia & adriana maria ribeiro] [In Process Citation] PMID- 11035495 TI - [Debate on the paper by dina czeresnia & adriana maria ribeiro] [In Process Citation] PMID- 11035496 TI - [Debate on the paper by dina czeresnia & adriana maria ribeiro] [In Process Citation] PMID- 11035497 TI - [Debate on the paper by dina czeresnia & adriana maria ribeiro] [In Process Citation] PMID- 11035498 TI - [Debate on the paper by dina czeresnia & adriana maria ribeiro] [In Process Citation] PMID- 11035499 TI - [Debate on the paper by dina czeresnia & adriana maria ribeiro] [In Process Citation] PMID- 11035501 TI - [The author replies] [In Process Citation] PMID- 11035500 TI - [Debate on the paper by dina czeresnia & adriana maria ribeiro] [In Process Citation] PMID- 11035502 TI - Environmental epidemiology applied to urban atmospheric pollution: a contribution from the Experimental Air Pollution Laboratory (LPAE). AB - Systematic investigation on the effects of human exposure to environmental pollution using scientific methodology only began in the 20th century as a consequence of several environmental accidents followed by an unexpected mortality increase above expected mortality and as a result of observational epidemiological and toxicological studies conducted on animals in developed countries. This article reports the experience of the Experimental Air Pollution Laboratory at the School of Medicine, University of Sao Paulo, concerning the respiratory system and pathophysiological mechanisms involved in responses to exposure to pollution using toxicological and experimental procedures, complemented by observational epidemiological studies conducted in the city of Sao Paulo. It also describes these epidemiological studies, pointing out that air pollution is harmful to public health, not only among susceptible groups but also in the general population, even when the concentration of pollutants is below the limits set by environmental legislation. The study provides valuable information to support the political and economic decision-making processes aimed at preserving the environment and enhancing quality of life. PMID- 11035503 TI - [Rotavirus infection in Brazil: epidemiology and challenges for its control]. AB - Worldwide, rotaviruses account for 600,000 to 870,000 deaths per year among infants and young children. In Brazil, rotaviruses were first seen in 1976 by scanning electron microscopy of stool samples from diarrheic infants in Belem, Para. Hospital-based studies have shown that rotaviruses are associated with 12 42% of cases of acute diarrhea. In addition, community-based studies yielded an average of 0.25 rotavirus-related diarrheal episodes per child per year. G types 1 to 4 account for about two-thirds of circulating strains, but the (unusual) P[8],G5 genotype has been claimed to cause over 10% of rotavirus diarrheal episodes. It has been shown that over 70% of children develop rotavirus antibodies by the age of 4-5 years. The tetravalent rhesus-human rotavirus vaccine (RRV-TV) conferred 35% protection according to a two-year follow-up study in Belem, Para, Brazil, but reached an efficacy of 60% during the first year of life. RRV-TV was also shown to be 75% protective against very severe gastroenteritis in northern Brazil. Vaccination with RRV-TV has been suspended recently in the United States because of the detection of intussusception as a side effect. Therefore, further vaccine trials in Brazil will probably involve rotavirus candidate vaccines other than RRV-TV. PMID- 11035504 TI - [Productive employment, gender and mental health]. AB - A cross-sectional study was conducted in Olinda, Pernambuco, to investigate a possible association between unemployment, informal work, and common mental disorders (CMD) assessed by the Self Reporting Questionnaire (SRQ - 20). While women working in the formal labor market showed significantly better mental health as compared to informal workers (OR = 3.02, 95% CI 1.3-7.2), housewives (OR = 2.29, 95% CI 1.0-5.0), and unemployed (OR = 2.66, 95% CI 1.1-6. 3) and inactive women (OR = 3.19, 95% CI 1.2-8.4), no difference was found among men. The actual pattern of the odds ratios suggests a modifying effect of gender in the association between employment status and CMD. However, the interaction term added to the final model was statistically significant for informal work, but not for unemployment. The results of the present study suggest that the experience of informal work may be different for men and women. This finding highlighted the need to incorporate a gender approach (reflecting a social dimension of sex related inequalities) to the theoretical framework based on social classes adopted here. PMID- 11035505 TI - [Intervertebral disk disease among oil drilling workers]. AB - A cross-sectional study among 1,026 oil drilling workers in Northeast Brazil found a prevalence rate of 5% for intervertebral disk disease, varying from 1.8% (activities without heavy lifting) and 4.5% (occasional lifting) to 7.2% (routine lifting). Disease prevalence was 10.5% among drilling workers with more than 15 years in the industry and 11.3% among those over 40 years of age. Prevalence ratio (PR) for the association between working in oil drilling operations and intervertebral disk disease was 2.3 (95% CI: 1.3-4.0). Retrospective information about exposure was collected to minimize the healthy worker survival effect. Using information on current occupation instead of occupational life history would cause an underestimated PR of 1.1 (95% CI: 0.6-1.9). Logistic regression showed results similar to the tabular analysis. Neither confounding nor interaction was evident. Growth of the Brazilian oil industry and recent changes in the work force contract and management, involving changes in risk management and health control, indicate a need for prompt ergonomic intervention in order to control intervertebral disk disease among oil drilling workers. PMID- 11035506 TI - [Mortality in childbearing-age women in Campinas, Sao Paulo (1985-1994)]. AB - To provide a profile of the main health problems in childbearing-age women, we studied all 3,086 death certificates from the SEADE Foundation for women from 10 to 49 years of age and residing in the municipality of Campinas, from January 1, 1985, to December 31, 1994. The primary cause of death was identified and classified according to the 10th review of the ICD. Population data were obtained from the Laboratory for Epidemiological Analyses and Research, UNICAMP. One fourth of the deaths were cardiovascular in origin, one-fifth were from external causes, and almost 20% were due to neoplasms. Maternal mortality was the ninth cause of death. External causes predominated in the 10-to-34-year age group, as compared to cardiovascular diseases and neoplasms in the 35-to-49-year group. Most alarming were the predominance of traffic accidents among causes of death in women up to age 34 (greater than AIDS during the study period) and the high mortality rate from homicides. PMID- 11035507 TI - Segmental hair mercury evaluation of a single family along the upper Madeira basin, Brazilian Amazon. AB - Mercury pollution (MeHg) up the aquatic food chains in the Amazonian ecosystems has been a major concern in environmental health. Riverside people (ribeirinhos) along the Upper Madeira river are heavy fish eaters. Hair is the best biomarker for MeHg exposure. By assuming a constant hair growth rate, it is possible to evaluate a temporal profile of Hg exposure over the recent defined past. In this paper we present the segmental total hair Hg concentrations from a single family from which some of the 10 persons investigated had high hair Hg concentrations (peak of 339 ppm). We also presented the hair MeHg content from 4 out of the 10 family members investigated. There was a wide variation in total hair Hg concentrations (8 to 339 ppm) among these individuals, who were mostly sharing their meals; there was also a wide variation in total Hg concentrations in the same individual over time (136 to 274 ppm). Hg speciation showed a mean and standard deviation in the MeHg content of 62% and 6%, respectively. The wide variation in total hair Hg concentration strongly indicated that it is possible to mitigate critical Hg exposure levels by conducting a fish advisory. PMID- 11035508 TI - [Economic and social impact of the National Tuberculosis Control Program (NTCP) on the Cuban population]. AB - Tuberculosis (TB) control strategies provide one of the greatest cost/effectiveness results. To assess the impact of the national TB control program on the Cuban population, the time series of new TB cases and death reports, potential years of life lost, and the numbers of beds for TB case hospitalization in the entire country during 1964-91 and 1992-96 were described by common simple calculation on the basis of estimated expected values. The reduction in new TB cases and deaths, potential years of life saved, and savings in expenditures for treatment, hospitalization, and unemployment compensation were estimated. From 1965 to 1991 new case reports were reduced by 94.6% (4% per year); 86,500 cases were avoided; 166,439 potential years of life were saved; 2,831,625 million pesos were saved on tuberculostatic drugs; 82.7 million pesos were saved on unemployment compensation under the social security system for workers with active TB. Estimated savings totaled 494,919,631.3 pesos. Nationwide intervention for TB control produced an important impact on the basis of the sociopolitical status making it possible to approach complete elimination of the disease in the future. PMID- 11035509 TI - [Socioeconomic factors and attitudes towards household prevention of American cutaneous leishmaniasis in an endemic area in Southern Bahia, Brazil]. AB - A survey was conducted to identify socioeconomic conditions and attitudes towards household prevention of American cutaneous leishmaniasis in Corte de Pedra, located in the county of Tancredo Neves, an endemic region in southern Bahia, Brazil. A questionnaire was applied in July 1997, focusing on social and economic variables, habits, and attitudes towards prevention of arthropod bites. All families (100%) living in the study area were surveyed, comprising 168 households with 851 individuals. Approximately 66.7% of the families earned up to one minimum wage, supporting an average of 5.1 residents per household. Most (57.2%) of the families did not use any type of protection against bites. Fumigation by burning various types of materials was the most customary form of prevention. Individual protection measures were rarely used. Since there was evidence of household and peridomiciliary transmission in the study area, use of impregnated bed nets is an alternative for intradomiciliary protection. PMID- 11035510 TI - [Age at menarche among schoolgirls from a rural community in Southeast Brazil]. AB - The purpose of this study was to determine the 3rd percentile (P3), 50th percentile (P50 = median age at menarche = MAM), and amplitude between the extremes (P97 and P3) of age at menarche among schoolgirls in the county of Barrinha, Sao Paulo, Brazil. Values were correlated with socioeconomic conditions such as social class, number of siblings, and father's employment status. This was a cross-sectional study based on the use of status quo adjusted by logit for calculation of percentiles. A questionnaire was applied to 1,602 schoolgirls aged 8 to 17 years (incomplete). MAM was 12 years (y) and 6 months (m), with a P97 of 10 y and 2 m and a P3 of 14 y and 10 m. Girls from lower-income families and those with unemployed fathers showed later MAM. No difference in MAM was observed with respect to number of siblings. Amplitude between P97 ad P3 was great in the overall sample. We conclude that Barrinha presented a MAM similar to and even lower than regional values for Brazil and for some developed countries. The study of the interval between extreme percentiles proved to be a better indicator of biological diversity and socioeconomic inequality than MAM alone. PMID- 11035511 TI - Dental caries in the primary dentition in public nursery school children in Juiz de Fora, Minas Gerais, Brazil. AB - The aim of this study was to assess the prevalence of dental caries in the primary dentition and associated variables in low socioeconomic preschool children enrolled in public nursery schools in Juiz de Fora, Minas Gerais, Brazil. Four public institutions were selected by geographic criteria (two in the central region and two in the peripheral region). The study population comprised 338 children (181 boys; 157 girls) aged 2-6 years old. Dental caries was recorded using the decayed, missing, and filled teeth (dmf-t) index. Among the examined children, 50.6% were caries-free. The mean dmf-t index was 2.03. It was higher in the peripheral nursery schools (p<0.01). A trend towards a difference between sexes (p = 0.06) was observed. Logistic regression analysis selected a previous child's visit to dentist (p<0.001), geographic location of the public nursery school (p<0.01), and age (p<0.01) as predictive variables for the dmf-t index. The study showed the need for an oral health program for this population, including both curative and preventive measures in order to achieve the WHO/FDI goals for the year 2000, namely 50% of children free of caries at age 5-6 years. PMID- 11035512 TI - [Planning and instrumental rationality: an analysis of theoretical production in strategic health planning in the 1990s in Brazil]. AB - This article analyzes recent theoretical production in Brazil on strategic health planning, focusing on instrumental logic. The approach centers on authors who have emphasized theoretical and methodological aspects of strategic planning. Emphasis is on the need to develop a new instrumentality capable of answering dilemmatic questions faced in health issues (professional health workers' efficacy/personal achievement) in order to recover the teleological planning action. The author provides a proposal that considers planning a modulator for the incorporation of technology by health institutions. PMID- 11035513 TI - A profile of sexually active male adolescent high school students in Lima, Peru. AB - To document knowledge and attitudes regarding sexuality and sexual practices of male adolescent high school students in Lima, Peru, a self-administered, anonymous survey was completed by 991 male adolescents aged 12-19 as part of a School-Based Sex Education Intervention model. Questions concerned sociodemographic information; family characteristics; personal activities; knowledge and attitudes regarding sexuality; sexual experience; and contraceptive use. Knowledge related to sexuality was limited. Males tended to mainly discuss sexuality with their male peers (49.8%). Attitudes towards sexual activity and condom use were largely positive, although some males expressed ambivalent feelings towards the latter. Of the sample, 43% had ever had sex; age at first sexual intercourse was 13 years. While 88% of the sample would use condoms, 74% also gave reasons for not using them. Sexual activity was related to age, ever having repeated a grade, living with only one parent or in a mixed family, activities such as going to parties, use of alcohol, tobacco, and drugs, and viewing pornographic videos or magazines. Many male adolescents were at risk of causing an unintended pregnancy or acquiring an STD. PMID- 11035514 TI - [Women caring for women: a study at the "Viva Maria" shelter, Porto Alegre, Rio Grande do Sul, Brazil]. AB - This research was conducted in Porto Alegre, Rio Grande do Sul, with a sample of battered women selected from a government shelter called the "Casa Viva Maria". We analyzed data on 110 women staying at the shelter during the previous two years (January 1996-June 1998). The profile of the women was as follows: abused women were young (mean age 29 years), all had low socioeconomic status, 12% were illiterate, 21% were black, 80% reported frequent abuse by their partners, and 18% had returned to violent homes. The researchers visited 34 former lodgers from the shelter and invited them to participate in a series of evaluation workshops. A total of 118 persons, including mothers and children, attended three evaluation meetings. During this process, researchers encouraged participants to express opinions, perceptions, and feelings about their past experience in the shelter and their own concept of violence. Finally, a focal group was organized with the "Viva Maria" staff members. Female workers reported how their job had been helpful for their personal development and had helped change their own lives. PMID- 11035515 TI - [Perspectives of chronically ill patients concerning medical care in Guadalajara, Mexico: a qualitative study]. AB - This paper reports partial findings from a broader study on the experience of people with chronic diseases. The objective was to explore the perspectives of diabetic patients towards medical care. A qualitative study was conducted in a poor neighborhood of Guadalajara, Mexico. Thirty subjects with diabetes mellitus participated in the study. Data was gathered by open and semi-open interviews in the subjects' homes and over the course of one year. Data were analyzed using a combination of content and conversational analyzing techniques. Three perspectives predominated when participants evaluated medical care: some define it as good, some as ambivalent, and the rest as bad. These perspectives were closely linked to their disease experience and available medical options according to their material resources. These perspectives change with time, are specific to each available service and type of medical care, and are constructed in terms of all the subjects' present chronic illnesses. Those treated through the social security system evaluate the care in negative terms, with the opposite occurring with those treated in public health care centers and private facilities. Implications regarding health care reform are discussed. PMID- 11035516 TI - [Avoidable perinatal deaths and obstetric health care structure in the public health care system: a case study in a city in greater metropolitan Rio de Janeiro]. AB - This paper investigates the occurrence of potentially avoidable perinatal deaths by associating failures in obstetric care with structural deficiencies in four maternity hospital comprising the local health care system in a city in Greater Metropolitan Rio de Janeiro in 1994. Information sources were a questionnaire, interview, observation, and hospital records. A theoretical organizational model was applied in data analysis. The four maternity hospitals showed problems in the three groups of factors used in this study: physical infrastructure, services, and equipment; organizational and administrative characteristics; and professional organization. In two maternity hospitals, the delay in cesarean deliveries was explained by deficiencies in the group of factors that includes facilities, services, and equipment. Health care failures directly associated with the physician were explained by high percentages of negative predisposing factors (mainly in the two private hospital), skill-related problems (more clearly in the two public hospitals), and by absence of measures for redirecting practice. PMID- 11035517 TI - [The evaluation of the physiological workload in the Brazilian legislation should be revised! The case of garbage collectors in Rio de Janeiro]. AB - The physiological workload (PW) involved in garbage collection was assessed in a probabilistic sample of 70 Rio de Janeiro city garbage collectors to determine the adequacy of Brazilian labor legislation regarding classification of work. PW was measured as energy expenditure (EE) and heart rate (HR) during total work time (TT) and actual time (AT) in garbage collection on 4 consecutive days. Median EE values were 288.4 and 319.1 kcal.h-1 during TT and AT, respectively, indicating moderately intense work according to Brazilian legislation. However, PW was considered heavy when work classifications based on individual response to work were used: 1) ratio of EE and resting metabolic rate was above 5.0, indicating heavy workload according to the WHO; 2) mean percentage of maximal EE was higher (36.2 and 41.1% for TT and AT, respectively) than the limit for garbage collection (30%) suggested as maximal for Dutch workers; and 3) percentage of maximal HR reserve was also higher than 30% (32.2 and 37.5% for TT and AT, respectively). These results indicate the need for a revision of the workload classification in the Brazilian legislation to take individual workers' characteristics into account. PMID- 11035518 TI - [Quality assessment of the public water supply in Nova Iguacu, Rio de Janeiro]. AB - This study proposes to develop methodologies for public water supply quality control by implementing evaluation and control mechanisms and enhancing environmental health surveillance decision-making. These objectives were based on data for waterborne diseases recorded at the Rio de Janeiro State Health Department. We selected the following neighborhoods: Posse (1st Township) and Caioaba (5th Township), with dissimilar situations, allowing us to produce a model of water supply coverage in the Municipality of Nova Iguacu, Rio de Janeiro State. The study underscored the poor quality of the water supply in the two townships, since 61% of the samples were positive on bacteriological examination, showing the undesirable effects of differential sanitation, especially in developing countries. PMID- 11035519 TI - [Economic crisis and infant mortality in Latin America since the 1980's]. AB - In the present study, based on data from nine Latin American countries, we found evidence of an association between the economic crisis and infant mortality during the last decades. The paper initially review previous studies on this issue and shows the need for a greater research focus on shorter time intervals. We then describe the deterioration and unequal conditions among the countries based on trends in selected social and economic indicators and the evolution of infant mortality rates. According to our statistical analysis, infant mortality bore an inverse association to short-term economic variations. We also found a significant and negative correlation between decreasing infant mortality rates and increasing poverty. The economic crisis displayed effects of varying intensity among the countries we analyzed, with social inequality appearing as the most probable explanatory variable. PMID- 11035520 TI - [Interaction between prescribers, dispensers, and patients: shared information as a possible therapeutic benefit]. AB - The article presents a critical view of the interaction between prescribers, dispensers, and patients, considering information one of the key issues in enhancing the qualitative aspects involved in this complex relationship. It describes the acquisition of information by health professionals, possible sources of this information, and the process involved in transforming it into knowledge. Briefly discussed are the physician's and pharmacist's roles, the patient's expectations as recipient, and consequences of pertinent health interventions. PMID- 11035521 TI - [An experience with public education in nutritional health in 2 communities in Jalisco, Mexico]. AB - This study describes a popular educational process conducted in two communities in Jalisco, Mexico. The purpose was to add an alfalfa concentrate to the population's diet as an alternative, locally available food source. Previous studies had shown that alfalfa contains high protein, vitamin, and essential amino acid levels and can be useful to supplement and improve child nutrition. This resource had not been used previously due to lack of knowledge concerning its properties and harvesting and processing procedures and because it had traditionally been used as livestock feed. The current study included four steps: 1) community knowledge, 2) a community survey using interviews, home visits, and child nutrition evaluation, 3) formation of work groups in a community meeting, and 4) an educational program, working with a self-diagnostic tool taking child nutritional status into account. Our work focused on two areas simultaneously: family nutrition and the alfalfa concentrate as a way to improve it. Although this process was lengthy, it resulted in the acceptance and inclusion of alfalfa concentrate. In addition, the community groups formed in the process remain as an ongoing organizational resource. PMID- 11035522 TI - [Evaluation of a comprehensive adolescent health care service]. AB - The authors evaluate the Adolescent Health Clinic in the Emaus community in Belem, Para, Brazil with regard to coverage, adequacy, accessibility, and utilization for the period 1994 to 1996. Coverage was calculated on the basis of clinical records and information collected from the target population. Adequacy was analyzed by comparing the service performance with goals established by PAHO/WHO for this kind of institution. Accessibility and utilization patterns were evaluated from information obtained by a population-based survey among adolescents living in the area. Results showed: a good degree of program adequacy vis-a-vis the final purpose, although some adjustments are needed, especially in human resources; lack of barriers to user access; reasonable coverage, despite the low proportion of consultations by adolescents; and utilization pattern compatible with the service provision profile, mainly directed towards curative and individual care. Recommendations are made to revise the hegemonic health care model to emphasize preventive, collective, and educational activities. PMID- 11035523 TI - [Diagnosis of intestinal amebiasis using coproscopic and immunological methods in a population sample in greater metropolitan Belem, Para, Brazil]. AB - We compare diagnostic methods for Entamoeba histolytica in fecal samples from the city of Belem, Para, Brazil. We analyze stool samples from children and adults (Group I); stool and serum samples from adults (Group II); and stool samples from children (Group III). In groups I and III, we used direct examination with lugol (DM), Faust et al (FM), and ELISA (detection of E. histolytica anti-GIAP coproantigen) and in group II, DM, iron hematoxylin staining (IHS), FM, ELISA, and the indirect immunofluorescence test (IFAT) for detection of IgG antibodies. Positivity was 10.50% by DM plus FM and 28.99% by ELISA. There was no correlation between positivity and age group. In Group II (n = 87), the positive rate was 4.59% by DM plus FM, 8.04% by IHS, 4.59% by IFAT, and 21.83% by ELISA. The ELISA test was the most sensitive for all groups. IFAT alone is still not a useful tool for diagnosis of E. histolytica infection. The ELISA test is simple, performed in one-third of cases used for IHS and IFAT, and greatly improves quality of diagnosis. We recommend this as the method of choice for diagnosis of suspected E. histolytica infection. PMID- 11035524 TI - [The culture of public health organizations: notes on the construction of an object]. AB - The author defines some central concepts for construction of the subject of "public health organizations". The concept of culture is exceedingly broad, and must be adjusted to the types of particular phenomena analyzed. As perceived in a public health organization, culture implies phenomena belonging to different social dimensions and which can be grasped through the concepts of political, civic, organizational, and professional culture. PMID- 11035525 TI - [Dengue study in a slum area in Rio de Janeiro: preliminary analysis]. AB - This study is part of a program to control and prevent dengue in a slum bordering on the grounds of the Oswaldo Cruz Foundation in Rio de Janeiro, Brazil. The results obtained from a questionnaire and entomological survey called attention to problems pertaining to the information transmitted by public health campaigns and its interpretation, since many practices result from misunderstanding or forgetting preventive messages. Dengue-related data include most frequent vector breeding sites, people's knowledge, and dengue-related habits. The study's conclusions in terms of dengue prevention point to the need for drafting messages not only about ideal preventive practices but also teaching possible solutions: disseminating frequent messages throughout the year and not only seasonally and planning health education to join health professionals and the population in the search for sustainable dengue control alternatives. PMID- 11035526 TI - [A research agenda for health policies and services in northeast Brazil]. AB - The aim of this paper is to gather elements for a discussion on health policies and services in northeast Brazil. The author begins with an analysis of the population's main health problems, the health system's limitations, and current opportunities for implementing health programs. She highlights the following research priorities: people's health situations according to their material living conditions and life styles; the private health sector's characteristics and development; the current decentralization process; and proposed innovations in services, programs, and systems. The author points out further that the policy making process in science and technology should assure a balance between researchers' freedom and people's health needs, regional focus, and universal issues. PMID- 11035528 TI - [Outsourcing in the Municipal Health Secretariat in Almirante Tamandare, Parana State, Brazil]. PMID- 11035527 TI - [New reproductive technologies: oocyte donation. What could be new in this field?]. AB - This article discusses the so-called "new" reproductive technologies. The author analyzes and challenges this adjective, since for over two decades this group of medical techniques and experiments has been widely disseminated in the medical market. The media's coverage of test tube babies, and especially developments in intervention on human germ cells and embryos, help challenge the supposed permanent novelty of everything surrounding reproductive technologies and genetic interventions. Society is doubtless experiencing an open process in full innovation, but the social and symbolic effects on planning maternity, paternity, and filiation are still not well perceived or discussed. To illustrate such contradictions, the article focuses on the case of oocyte donation, highlighting the need for new perspectives in terms of social control over the dissemination of reproductive technologies. PMID- 11035529 TI - [The dynamics of territorial occupation and visceral leismaniasis in the city of Sao Luis, Maranhao state, Brazil]. PMID- 11035530 TI - [Health and nutritional conditions of indigenous and non-indigenous riverine children along the rio Solimoes, Amazonas state, Brazil]. PMID- 11035531 TI - Influence of pigment content, intracellular calcium and cyclic AMP on the ability of human retinal pigment epithelial cells to contract collagen gels. AB - PURPOSE: The aim of the study was to determine to what extent collagen gel contraction could be reduced by calcium and calmodulin antagonists and agents that elevate cyclic AMP in order to develop a pharmacological approach to prevent/arrest RPE contraction of epiretinal membranes in proliferative vitreoretinopathy. We also explored a possible role of pigment in collagen gel contraction. METHOD: We measured RPE mediated contraction of 3D collagen gels in the presence and absence of the calcium and calmodulin antagonists TMB8, Verapamil and Tamoxifen and the cAMP elevating agents IBMX and Forskolin. The effect of pigment on collagen gel contraction was assessed by comparing gel contraction mediated by RPE cells re-pigmented with melanin with that mediated by unpigmented RPE. The effect of IBMX on RPE proliferation was assessed using a BrdU ELISA and the effects of IBMX on RPE cytoskeleton and cell shape were assessed using Actin and Cytokeratin immunocytochemistry. RESULTS: We report that both cAMP elevating agents and calcium and calmodulin antagonists reduce RPE mediated collagen gel contraction. Cyclic AMP elevation was more effective than a reduction in calcium in reducing contraction. There were no significant advantages in combining both approaches. The presence of melanin had no effect on gel contraction. Calcium antagonists and particularly agents which elevate cAMP caused RPE cells in collagen gels to extend fewer and shorter processes. cAMP elevation in particular caused RPE cells to become more rounded and develop arborized cell processes. Immunostaining for actin and cytokeratin revealed changes in cytoskeletal organisation in response to IBMX in that cells contained less actin than untreated cells and concentrated cytokeratins more centrally. CONCLUSION: We have identified two possible pharmacological approaches which may provide a new direction for preventing or slowing down the development of PVR. PMID- 11035532 TI - The local and systemic secretion of the pro-inflammatory cytokine interleukin-6 after transplantation of retinal pigment epithelium cells in a rabbit model. AB - The rejection of retinal pigment epithelium (RPE) allografts is one of the major problems for long-term success after retinal transplantation. However, the details of the immunological interactions in the subretinal space after transplantation are still unknown. The aim of our study was to investigate the role of IL-6 in the rejection process in the subretinal space and to use IL-6 monitoring for a possible early sign of rejection after transplantation of allogeneic RPE cells. For this we used a model of transplanting pigmented RPE cells, either activated in vitro with 1000 U/ml interferon-gamma (IFN-gamma) for 8 days or non-activated, into 30 albino rabbits. The IL-6 was investigated 3, 5, 7, 9 and 14 days after transplantation. Additionally, sham operated animals and the untreated eyes served as controls. At these time-points the animals were killed, the liquid in the vitreous cavity and serum was collected and the IL-6 present in these samples was quantified with an enzyme-linked immunosorbent assay. Under these conditions, IL-6 was detected in the liquid of the vitreous cavity and in the serum of all RPE-transplanted rabbits. In the group receiving activated RPE two cytokine peaks were measured, 3 and 7 days after transplantation in the vitreous cavity. In non-activated grafts, a maximum was detected on the 5th day after transplantation. Generally, the detected quantity of IL-6 depended on the host status and on the phase of rejection. No significant changes were seen in the sera from either group. Possibly, the host RPE cells are the main source of this interleukin in the transplantation area. The measuring of IL-6 in the rejection model suggests that it plays a role in the immune cascade in the subretinal space. PMID- 11035533 TI - Pupillography as an objective indicator of fatigue. AB - PURPOSE: To determine whether parameters calculated from pupillary activity can identify subjects with sleep deprivation, and whether the objective values correlate with a subjective feeling of fatigue. METHODS: pupil size in the dark was recorded continuously for 10 minutes in 12 healthy volunteers using an infrared video camera. Two recordings were made for each subject: after a full night's sleep, and after 24 hours of sleep deprivation. Several parameters calculated from pupil size and activity were analyzed and compared with a subjective rating of the state of alertness provided by the participants in each test. RESULTS: All pupillary parameters differed significantly between alertness and fatigue (p = 0.0076-0. 0186). Changes in one of the parameters - average pupillary diameter - correlated with changes in the subjective level of sleepiness (r = -0.51, p = 0.028). Although the values of most parameters differed among subjects, an absolute value of more than 25 in one parameter, cumulative pupillary variability ratio, was always associated with sleep deprivation. CONCLUSION: On-line analysis of the pupillogram using the suggested parameters can be performed easily to produce a real-time assessment of an individual's state of alertness or fatigue that correlates with his/her subjective assessment of this state. PMID- 11035534 TI - Effects of inhibition of glycation and oxidative stress on the development of cataract and retinal vessel abnormalities in diabetic rats. AB - PURPOSE: To study effects of inhibition of glycation, and oxidative stress on the development of cataract and retinal vessel abnormalities in diabetic rats. METHODS: Diabetes was induced in male Wistar rats with streptozocin (STZ; 60 mg/kg BW, i.p.). Diabetic as well as strain matched control rats were fed 1) a normal diet, 2) addition of aminoguanidine in the drinking water (0.5 g/l for diabetic rats and 1.0 g/l for control rats) or 3) probucol in the pellets (1% w/w). After 6 months, the number of acellular vessels, endothelial cells and pericytes were counted in trypsin digested retinal vessel preparations, and the total retinal tissue amount of glutathione (GSH) and cysteine was measured with HPLC. RESULTS: Cataract formation occurred after 13 weeks in diabetic animals compared with 17 weeks for those treated with aminoguanidine, and 16 weeks for those treated with probucol (p < 0.001 in both cases). Aminoguanidine inhibited the formation of acellular collapsed capillary strands, 9 (3-14) vs. 18 (12-262) (median, range) per quadrant in untreated diabetic rats (p = 0.004), while probucol did not have any effect. Neither aminoguanidine, nor probucol influenced the endothelial/pericyte ratio. Diabetes caused a reduction in the GSH/cysteine ratio (10.7 +/- 0.6 vs. 15.3 +/- 1. 5) (mean +/- SD; p < 0.001). Probucol partly restored this imbalance (p < 0.05) whereas aminoguanidine did not. CONCLUSIONS: The results indicate that cataract formation in diabetes involves both glycation and oxidative stress processes. The reduced formation of acellular collapsed capillary strands by aminoguanidine suggests a potential role for glycation in vascular damage. The positive effect of probucol on cysteine/GSH metabolism imbalance indicates that derangements of one of the retinal defense systems against oxidative stress can be normalized by antioxidants. PMID- 11035535 TI - Conjunctival inflammation induces Langerhans cell migration into the cornea. AB - PURPOSE: The virtual absence of Langerhans cells (LC) in donor or recipient corneal epithelium is known to be an important factor in the acceptance of orthotopic corneal allografts. Though it is well known that various types of stimulation to the cornea induce LC migration into the corneal epithelium, resulting in poor graft survival, the influence of conjunctival inflammation on LC migration into the cornea has not been elucidated. Therefore we examined whether LCs migrate into the cornea in the presence of conjunctival inflammation. METHODS: Sixteen BALB/c mice were divided into four groups. Group A: 4 mice with corneal inflammation induced by two 9-0 silk interrupted sutures in the central cornea (positive control); Group B: 4 mice with conjunctival inflammation induced by two 9-0 silk interrupted sutures in the temporal and nasal bulbar conjunctiva 1 mm from the limbus; Group C: 4 mice with conjunctival inflammation by two 10-0 nylon interrupted sutures in the temporal and nasal bulbar conjunctiva 1 mm from the limbus; and Group N: 4 mice with no inflammation (untreated, naive control). Fourteen days after suturing, the mice were sacrificed and LCs migrated into the corneal epithelium were counted by immunofluorescence assay using anti-Ia antibody. RESULTS: In Group A, Ia(+) cells in the cornea totaled 29.4 +/- 3.8 cells/mm(2); in Group B, 7.9 +/- 1.2 cells/mm(2); in Group C, 7.8 +/- 0.7 cells/mm(2); and in Group N, 1. 6 +/- 0.5 cells/mm(2). Significantly greater numbers of Ia(+) cells were detected in Groups A, B and C than in Group N (p < 0.005). CONCLUSIONS: Conjunctival inflammation caused by sutures in the bulbar conjunctiva induced LC migration into the cornea. These results indicate that conjunctival inflammation influences the corneal immunological environment, and may affect the fate of orthotopic corneal allografts. PMID- 11035536 TI - Lipoxygenase metabolism following laser induced retinal injury in rabbits. AB - PURPOSE: 1) to investigate whether leukotriene B(4) (LTB(4)) is a factor in the inflammatory reaction following chorioretinal laser injury in rabbits; 2) to study its relationship with the cyclooxygenase (COX) metabolic pathway; 3) to study the influence of Nordihydroguaiaretic acid (NDGA), an inhibitor; of the lipoxygenase (LOX) cascade, on both COX and LOX metabolism. METHODS: Prostaglandin E(2) (PGE(2)) and LTB(4) synthesis by incubated samples of chorioretina obtained from rabbits' eyes exposed to Neodymium:Yag laser along with these eicosanoids accumulation in the vitreous were measured over one week follow-up period. The effect of NDGA pre-treatment on the COX and the LOX pathways in the laser-injured chorioretina was also assessed. PGE(2) and LTB(4) levels in the vitreous and in the chorioretina incubation medium were quantified using the radioimmunoassay technique with the appropriate antibodies. RESULTS: LTB(4) in vitro production by rabbits' chorioretina subjected to ND; YAG laser was significantly elevated compared to control, peaking on day 7 to levels 2.45 fold greater than baseline (p < 0.01). PGE(2) formation, following a different pattern, was also enhanced and its maximal level (5.2 fold higher than control, p < 0.01) was achieved at the initial phase (day 1 post laser). Laser irradiation caused also an increase in the two eicosanoids accumulation in the vitreous, which was however not proportional to their production levels. NDGA treatment was associated with a sustained decrease in LTB(4) content in the vitreous, but had no effect on PGE(2) vitreal levels. CONCLUSIONS: Laser irradiation of the rabbits' retina induces an alteration in the LOX metabolic pathway, which is dissociated from the influence on the COX cascade, pointing for the first time to a possible role played by LTB( 4) as a mediator in the chorioretinal inflammatory reaction, with no connection to the role played by PGE(2). NDGA selectively inhibited LOX activity without affecting COX activity. PMID- 11035537 TI - Evaluation of fluorescein-labeled autologous leukocytes for examination of retinal circulation in humans. AB - PURPOSE: Increased leukocyte-endothelium interaction have been suggested as a phenomenon contributing to capillary occlusion and/or rupture of the blood-retina barrier during human retinal vascular diseases. This study was performed to evaluate if fluorescein-labeled autologous leukocytes (FLALs) can be used for examination of leukocyte transit in the human retina. METHODS: The preparation consisted of human dextran-separated leukocytes mixed with fluorescein. After reinjection in normal subjects and in one diabetic patient, a confocal scanning laser ophthalmoscope was used to visualize them in the retinal circulation. The changes between FLALs and control leukocytes in the expression of leukocytes adhesion molecules CD11b and CD62L were evaluated by flow cytometry. RESULTS: The circulating FLALs were clearly visible in retinal vessels. The mean (+/- SD) capillaries velocity was 1.43 (+/- 1.3) mm/s in the macula and 1.82 (+/- 1.4) mm/s in the peripapillary area. No leukostasis was detected in the normal subjects, while it was detected in te diabetic patient. Flow cytometry revealed an increase in CD11b and a decrease in CD62L expression of leukocytes after labeling, suggesting that compared to normal leukocytes FLALs are more susceptible to interact with vascular endothelium. CONCLUSIONS: The use of FLAL is presently the only technique applicable in humans for study of leukocyte transit in the retina. Their preparation is technically simple and unexpensive. Precise measurement of the velocity of leukocytes in small vessels can be obtained. Despite evidence of a certain degree of leukocyte activation after the labeling procedure, no leukostasis was detected in vivo in normal subjects. Potential applications for this technique may include the detection of leukostasis in the human retina during severe forms of diabetes and retinal phlebitis. PMID- 11035538 TI - Ocular penetration of topical Delta9-tetrahydrocannabinol from rabbit corneal or cul-de-sac application site. AB - PURPOSE: To determine the relative effects of low-volume drop placement either on the corneal surface or in the lower cul-de-sac on Delta9-tetrahydrocannabinol (THC) penetration into ocular fluids and tissues. METHODS: Pigmented rabbits received 10 microl drops in one eye at either application site. Care was taken to ensure that the drop remained at the site of drug placement. At different intervals after drop administration containing ( 3)H-Delta9-tetrahydrocannabinol in combination with either alpha-cyclodextrin or hydroxypropylmethylcellulose as excipients in different vehicles, eye fluids and tissues were harvested, digested, sampled and counted. RESULTS: Greater quantities of THC were found in the corneal epithelium, stroma/endothelium and aqueous humor of treated eyes after corneal drug application relative to values obtained after cul-de-sac drug application. Cul-de-sac applications gave higher values for inferior bulbar conjunctiva compared with corneal application. These results occurred for each vehicle despite a quantitative difference in drug penetration between the two vehicles. No THC was found in untreated contralateral eyes. CONCLUSIONS: Application of a lipophilic drug to the rabbit corneal surface exposed in the interpalpebral space produced higher tissue and fluid drug concentrations in the anterior segment of the eye compared with drug application into the lower cul-de sac. This observation has relevance to pre-clinical drug toxicity testing as well as drop placement in patients for the treatment of ocular diseases. PMID- 11035540 TI - Changes in diabetic retinal matrix protein mRNA levels in a common transgenic mouse strain. AB - Recently, all the structural features of non-proliferative diabetic retinopathy have been demonstrated in mice fed 30% galactose for 21-26 months. To determine whether changes in retinal matrix protein mRNA levels occur early in the course of murine diabetes we used a competitive RT-PCR method to quantitate retinal mRNA levels in an inbred mouse strain (FVB) commonly used for transgenic studies. Retinal mRNA was prepared from STZ-diabetic and non-diabetic FVB mice at 4, 8, 12 and 16 weeks and cDNA encoding basement membrane components was quantitated using MIMIC constructs that compete for the same primer pairs. alpha1 (IV) collagen, the beta1 and gamma1 chains of laminin, fibronectin, and vitronectin mRNAs were quantitated. For alpha1 (IV) collagen, statistically significant diabetes-induced increases were apparent by 8 weeks (3.11 +/- 0.20 vs. 1.29 +/- 0.19 x 10(6) molecules/mg total RNA, p < 0.005). Similarly, diabetes-induced increases were observed by 8 weeks for the beta1 chain of laminin (4.54 +/- 0.22 vs. 1.85 +/- 0.43 x 10(5) molecules/mg total RNA, p < 0.005), the gamma1 chain of laminin (7. 33 +/- 0.29 vs. 4.84 +/- 0.76 x 10(4)/microg total RNA, p < 0.05), and for fibronectin (2.22 +/- 0.21 vs. 1.35 +/- 0.15 x 10(6) molecules/mg total RNA, p < 0.05). The magnitude of change was greatest for alpha1 (IV) collagen (2.4-fold) and beta1 laminin (2. 5-fold) at 8 weeks, and least for fibronectin (1.6-fold). A smaller diabetes-induced increase in vitro nectin mRNA was also observed, but it failed to reach statistical significance at 12 and 16 weeks. These data provide the basis for assessing the effects of genetic manipulation on diabetic retinopathy in transgenic mouse models. PMID- 11035539 TI - Involvement of calpain isoforms in ischemia-reperfusion injury in rat retina. AB - PURPOSE: Much evidence has accumulated suggesting that activation of calpain causes neuronal cell death in ischemic brain. However, little is known about the involvement of calpain in retinal cell death in ischemic injury. Thus, the purpose of present study was to investigate the involvement of calpain isoforms (m- and mu-calpain) in ischemia-reperfusion injury in retina from rat. METHODS: Retinal ischemia was produced by occlusion of the central retinal artery for one hour, and this was followed by reperfusion for seven days. Calpain mRNAs, calpain activities, total calcium content and proteolysis of alpha-spectrin were determined in retina. Effect of a calpain inhibitor SJA6017 was histologically tested in retinal injury after ischemia-reperfusion. RESULTS: Following retinal ischemia, most of cells in the ganglion cell layer were sloughed off by day 1 after reperfusion, followed by loss of cells in the inner plexiform layer on day 3 and loss of cells in the inner nuclear layer by day 5. These morphologic changes were accompanied by several presumptive biochemical indicators of calpain activation: increased calcium, proteolysis of alpha-spectrin (a sensitive substrate for calpains), decreased caseinolytic activity for both calpains (suggesting calpain activation followed by autolytic degradation), increased mRNA levels for mu-calpain and calpastatin - the endogenous inhibitor of calpains - and decreased mRNA levels for mu-calpain. Moreover, the calpain inhibitor SJA6017 protected the reduction of cell density in the ganglion cell layer after ischemia reperfusion. CONCLUSION: These results suggest that calpain isoforms may play an important role in neuronal cell death induced by retinal ischemia-reperfusion injury in rat. PMID- 11035541 TI - AlphaB-crystallin in lacrimal gland duct and tears. AB - PURPOSE: To investigate alphaB-Crystallin expression and localization in the lacrimal gland and tear fluid. METHODS: Mouse, rat, porcine, monkey and human lacrimal gland samples were immuno-histochemically and immuno-electron microscopically stained with various antibodies against alphaB-crystallin. Western- and Northern-blotting was performed to demonstrate the presence of alphaB-crystallin mRNA and protein. Human tear fluid was analyzed for the presence of alphaB-crystallin using dot blotting. RESULTS: alphaB-Crystallin is located in the lacrimal gland duct cells but not in the acini. Electron microscopically, the protein was frequently found in apical electron-dense granules of lacrimal duct cells, occasionally also in the duct lumina. Western blotting confirmed the presence of alphaB-crystallin in the lacrimal gland, Northern blot samples revealed the presence of alphaB-crystallin mRNA. In the human tear fluid, alphaB-crystallin was present in all samples investigated. CONCLUSIONS: We demonstrate for the first time that alphaB-crystallin is present in the lacrimal gland. Presence of the protein in apical secretory granules as well as presence in the tear fluid might indicate secretion of alphaB-crystallin into the tear fluid. PMID- 11035542 TI - On the value-neutrality of the concepts of health and disease: unto the breach again. AB - A number of philosophers of medicine have attempted to provide analyses of health and disease in which the role that values play in those concepts is restricted. There are three ways in which values can be restricted in the concepts of health and disease. They can be: (i) eliminated, (ii) tamed or (iii) corralled. These three approaches correspond, respectively, to the work of Boorse, Lennox, and Wakefield. The concern of each of these authors is that if unrestricted values are allowed to infect our concepts of health and disease, then anything could be construed as healthy or diseased. They believe that, if at all possible, such a result should be avoided. Unfortunately, as I argue, this result is unavoidable and such attempts to limit values in these concepts are destined to fail. I argue for this position by showing how each of these three attempts to provide value restricted analyses of health and disease fail as analyses of the concepts of health and disease and that they fail because of their attempts to restrict the role of values in their accounts. I also show how, despite their best efforts, each of these analyses are, themselves, value-driven and value-laden. This leads to the conclusion that values infect our concepts of health and disease at all levels. PMID- 11035543 TI - The ends of medical intervention and the demarcation of the normal from the pathological. AB - This study examines the ends of medical intervention and argues that mainstream contemporary medicine assumes that appropriate ends may be discovered (i.e., naturalism), rather than created or decided upon (i.e., conventionalism). The essay then applies these considerations to the problem of the demarcation of the normal from the pathological. I argue that the common formulations of this dispute commit a fallacy, as they characterize the "normal" as a state of the organism and not as an ongoing process within it. Such a process may be characterized as self-creation and self-repair. Such considerations support the conclusion that normality may be regarded as a regulative idea, rather than as an end-state, and as part of the ends of medical intervention, depending upon choice and context. PMID- 11035544 TI - Fuzzy health, illness, and disease. AB - The notions of health, illness, and disease are fuzzy-theoretically analyzed. They present themselves as non-Aristotelian concepts violating basic principles of classical logic. A recursive scheme for defining the controversial notion of disease is proposed that also supports a concept of fuzzy disease. A sketch is given of the prototype resemblance theory of disease. PMID- 11035545 TI - On the place of fuzzy health in medical theory. AB - This commentary on Sadegh-Zadeh's article 'Fuzzy health, illness and disease,' has its focus on the philosophical background for applying fuzzy logic to medical theory. I concentrate on four issues. First, I contest some of Sadegh-Zadeh's statements on the present state of the theory of medicine, in particular with regard to assumptions ascribed to contemporary theorists. Second, I consider Sadegh-Zadeh's interesting idea that a person can have a disease to varying degrees, from not having it at all to having it completely. I argue that there are difficulties pertaining to the definition of particular diseases, which obstruct the application of this idea. The following two points concern medical semantics and principles for definition in general. I take issue with Sadegh Zadeh's description of the correct procedure for definition. I also contest his unconditional proposal for a social definition of health, illness and disease. PMID- 11035547 TI - Optic atrophy in association with cobalamin C (cblC) disease. AB - PURPOSE: To report the association of optic atrophy with cobalamin C (cblC) disease. METHODS: Descriptive case reports on three patients, two of whom were siblings. RESULTS: All three patients with cblC disease exhibited bilateral optic atrophy with decreased visual acuity. Of the two siblings, the younger sister had received cobalamin supplements from birth and the mother had been given cobalamin supplements prenatally. CONCLUSION: These three cases confirm the association of optic atrophy with cblC disease. Early treatment with cobalamin supplements does not appear to prevent the development of optic atrophy. PMID- 11035546 TI - Mutational analysis and clinical correlation in Leber congenital amaurosis. AB - Leber congenital amaurosis (LCA, MIM 204001) is a clinically and genetically heterogeneous retinal disorder characterized by severe visual loss from birth, nystagmus, poor pupillary reflexes, retinal pigmentary or atrophic changes, and a markedly diminished electroretinogram (ERG). PURPOSE: To examine 100 consecutive patients with LCA in order to assess the relative burden of the three known genes involved in LCA, namely retinal guanylyl cyclase (GUCY2D), retinal pigment epithelium protein ( RPE65), and the cone-rod homeobox (CRX), and to define their clinical correlates. METHODS: Mutational analysis and detailed clinical examinations were performed in patients diagnosed with LCA at the Johns Hopkins Center for Hereditary Eye Diseases and the Montreal Children's Hospital. RESULTS: Mutations were identified in 11% of our patients: GUCY2D mutations accounted for 6%, while RPE65 and CRX gene mutations accounted for 3% and 2%, respectively. The clinical presentation was variable; however, the visual evolution in patients with mutations in GUCY2D and CRX remained stable, while individuals with mutations in the RPE65 gene showed progressive visual loss. CONCLUSIONS: This study suggests that molecular diagnosis of Leber congenital amaurosis could provide important information concerning prognosis and course of treatment. PMID- 11035548 TI - Expression of the TIGR gene in the iris, ciliary body, and trabecular meshwork of the human eye. AB - Mutations in the the glaucoma gene GCL1A, also known as trabecular meshwork glucocorticoid response (TIGR) or myocilin (Myoc), have been shown to be associated with juvenile-onset primary open-angle glaucoma. Very little is known about the pattern of expression of the TIGR gene in human ocular tissues. In-situ hybridization experiments demonstrated the localization of TIGR mRNA in cells throughout the iris, ciliary muscle, and the filtering portion of the trabecular meshwork of normal eye donors. The expression of TIGR protein was investigated by Western blot using an epitope-directed antibody to the carboxy terminus region of TIGR. This antibody was able to distinguish a recombinant TIGR fusion protein from a truncated TIGR form containing the naturally occurring Gln(368)-->stop mutation. In tissue extracts from the iris, ciliary body, and trabecular meshwork, the antibody recognized a major protein band of 57-kDa molecular mass. Deglycosylation treatment with PNGase F, NANase II, and O-glycosidase indicated that the 57-kDa protein in these tissues was unglycosylated. In agreement with this observation, in coupled in-vitro transcription/translation systems, the 57 kDa TIGR protein was unaffected by the presence of the processing and glycosylation activities of canine pancreatic microsomal membranes. These findings support the view that the expression of TIGR mRNA in cells of the iris, ciliary body, and trabecular meshwork correlates with that of TIGR protein, and that the 57-kDa TIGR protein was unglycosylated. These results, which are in contrast with earlier reports, raise the possibility that the TIGR protein might be processed into distinct forms in a tissue-specific manner. PMID- 11035549 TI - Clinical characteristics of 14 japanese patients with X-linked juvenile retinoschisis associated with XLRS1 mutation. AB - To characterize the clinical features associated with XLRS1 gene mutations in Japanese patients with X-linked juvenile retinoschisis (xlRS), we evaluated the following data on 14 Japanese males from 13 unrelated families with XLRS1 mutations: age and symptoms at first visit to an ophthalmologist and ophthalmologic findings including visual acuity, refractive errors, fundoscopic appearance, and results of electroretinography (ERG) and electro-oculography (EOG). Each clinical finding was reviewed when the patients were between six and eight years of age. The best-corrected visual acuity in 12 patients (24 eyes) between the ages 6 and 8 years ranged from 1.0 to no light perception. Macular abnormalities were present in all cases. Peripheral retinoschisis was present in 14 of 26 eyes (53.8%). In the 21 eyes for which a single-flash ERG had been recorded, b-wave amplitude was reduced in 17 eyes. The EOG showed a low Arden ratio in three of the 13 eyes in the seven patients evaluated. No clear relationship was observed between the clinical features and the existing mutations. Three of four patients with a visual acuity less than 0.1 had retinal detachment or severe macular lesion that had occurred before the age of four years. Two patients harbored deletions of exon 1 or of the boundary region between exon 3 and intron 3, and one patient harbored R182C in exon 6. The present study shows a heterogeneity of mutations in the XLRS1 gene and phenotypic variations in 14 Japanese patients with xlRS. PMID- 11035550 TI - Bilateral papillomacular retinal folds and posterior microphthalmus: new features of a recently established disease. AB - Clinical findings of a 2.5-year-old girl presenting with barely detectable horizontal nystagmus and high hypermetropia are described. Despite the normal appearing anterior segments, the child had posterior microphthalmus and bilateral papillomacular retinal folds, conforming to a recently described, rare congenital disease. The patient also had significant posterior pole excyclorotation and avascular zones at the extreme temporal periphery without ridge formation or neovascularization. These findings were not reported previously. Other remarkable features include mildly depressed photopic and scotopic electroretinogram amplitudes and a short axial length of the vitreous cavity compared to age matched normals, measured by ultrasonography. The present case adds new elements to this relatively rare ocular developmental abnormality. PMID- 11035551 TI - Choroideremia, sensorineural deafness, and primary ovarian failure in a woman with a balanced X-4 translocation. AB - We present clinical and cytogenetic studies of a female patient affected with choroideremia, mild sensorineural deafness, and primary amenorrhea showing a balanced translocation between chromosomes X and 4. The breakpoint was precisely defined applying FISH techniques: 46,X,t(X;4)(q21.2;p16.3).ish t(X;4)(D4S96+, D4F26+; wcpX+). The X-chromosomal breakpoint was located within a region where both the choroideremia locus and a deafness locus (DFN3/POU3F4) have been mapped. The presence of X-linked disorders in this balanced carrier of X-autosomal translocations (XAT) can be explained either by the disruption of the structural coding or regulatory sequences of the gene(s) or by the submicroscopic deletion of this region leading to a contiguous gene deletion syndrome. The primary ovarian failure (POF) found in the present case has been already observed in XAT when the breakpoint is within a previously defined critical region (Xq13-26). A position effect is postulated as a possible explanation. PMID- 11035552 TI - Ultraviolet light exposure as a risk factor for ocular melanoma in Queensland, Australia. AB - The state of Queensland, Australia, has one of the highest incidences of cutaneous melanoma in the world; this has been linked to the high sun exposure of the mainly Caucasian population. The role of sun exposure in the development of ocular melanoma (melanoma of the conjunctiva, iris, ciliary body or choroid) remains unclear. A case-control study involving 125 patients with ocular melanoma treated between 1972 and 1996, and 375 age- and sex-matched controls (three for each patient) was performed. A standardised telephone questionnaire examining ultraviolet exposure and other potential risk factors was administered. Cumulative lifetime ocular ultraviolet B (UV-B) exposure was assessed using the Melbourne Visual Impairment Project instrument. Risk factors identified include personal history of melanoma of the skin (odds ratio [OR] 2. 42, 95% confidence interval [CI] 0.88, 6.62) or other skin cancers (OR 1.52, CI 0.99, 2.35), and family history of ocu-lar melanoma (OR 6.89, CI 0.70, 67.38). Protective factors included olive or black skin (OR 0.72, CI 0.40, 1.31), brown iris colour (OR 0.89, CI 0.51, 1.54), high resistance to sunburn (OR 0.58, CI 0.26, 1.31), and wearing prescription glasses (OR 0.78, CI 0.48, 1.25). Sunglass wearing was not found to be protective. Cumulative lifetime ocular UV-B exposure was not found to be a risk factor for ocular melanoma. However, there were too few cases of conjunctival and iris melanoma for these to be analysed as separate sub-groups. PMID- 11035553 TI - Utility-based estimates of the relative morbidity of visual impairment and angina. AB - PURPOSE: To quantify and compare the reduction in quality of life due to visual impairment and angina using patient preferences (utilities). METHODS: Using a standard time tradeoff method, we obtained utilities for current vision, monocular and binocular blindness, current angina, and moderate angina in 60 patients with both vision problems and angina pectoris who sought care at the National Eye Institute (NEI), National Naval Medical Center, or Barnes-Jewish Hospital. Patients were characterized clinically based on visual acuity and the Duke Activity Status Index (DASI). Patients also completed a seven-item version of the NEI Visual Functioning Questionnaire and the SF-36 Health Survey Questionnaire. RESULTS: Patients had a median visual acuity of 20/100 in the worst eye, 20/40 in the better eye, and a median DASI of 24.2 (0 = severe functional limitations due to anginal symptoms, 58.2 = no limitations). There was substantial variation in utilities among patients. The average utility for current vision (relative to ideal vision [= 1.0] and death [= 0.0]) was 0.82; the average utility for current angina (relative to no angina symptoms [= 1.0] and death [ = 0.0]) was 0.89. Among 26 patients with both visual impairment and recent anginal symptoms, the decrement in utility (on a scale ranging from ideal health [= 1.0] to death [= 0.0]) imposed by current visual impairment was greater than that imposed by current angina symptoms (0.146 versus 0.072, p=0.08, Wilcoxon signed rank test). The decrement in utility associated with binocular blindness was greater than the decrement associated with the symptoms of moderate angina (0.477 versus 0.039, p<0.0001). CONCLUSIONS: Clinical status is not a surrogate for patient preferences regarding vision impairment or angina. There is substantial variation in utilities within the study population for both experienced and theoretical impairment states which is not explained by variations in clinical status. Some states of visual impairment may pose a greater quality of life burden than anginal symptoms. Because patient preferences for vision vary greatly, individual assessment is warranted for consideration in therapeutic decision making. PMID- 11035554 TI - Astigmatism and Amblyopia among Native American Children (AANAC): design and methods. AB - The overall goal of the AANAC study is to improve detection of astigmatism and prevention of amblyopia in populations with a high prevalence of astigmatism. To meet this goal, the study will evaluate four methods of screening for astigmatism in preschool children and will assess both the short-term and long-term benefits of early correction of astigmatism in improving acuity and preventing amblyopia. This paper presents an overview of the design and methodology of the AANAC study. Subjects are members of the Tohono O'Odham Nation, a Native American tribe with a high prevalence of astigmatism. Preschool-age children who attend Head Start are screened with four tools: the Marco Nidek KM-500 autokeratometer, the MTI photoscreener, the Nikon Retinomax K-Plus autorefractor, and the Lea Symbols acuity chart. Sensitivity and specificity for detection of significant astigmatism, as measured by a technique that uses both cycloplegic retinoscopy and cycloplegic autorefraction, is determined for each of the four screening tools. Presence of amblyopia is evaluated by measurement of best-corrected recognition acuity and acuity for orthogonal gratings. Spectacles are provided to all 3-year-old children with > or =2.00 diopters (D) of astigmatism and all 4- and 5-year-old children with > or =1.50 D of astigmatism. Persistence of amblyopia after glasses wearing is evaluated by follow-up measurement of best corrected recognition acuity and acuity for orthogonal gratings, conducted 2-5 months after glasses are prescribed. Long-term effectiveness of early screening and glasses prescription is evaluated through measurement of recognition acuity in two groups of first-grade children: one group who participated in the Head Start program before the intensive vision screening program was initiated, and a second group who participated in the study's Head Start vision screening program. PMID- 11035555 TI - Recruitment methods for community-based screening for diabetic retinopathy. AB - Regular screening of all people with diabetes is the most efficient and cost effective way to detect early stages of diabetic retinopathy so that laser treatment can be performed at the optimal time. A major aim of the Program for the Early Detection of Diabetic Retinopathy was to increase compliance with guidelines for screening for diabetic retinopathy. This community-based screening program used non-mydriatic retinal photography and was initiated in four areas of Victoria, Australia from 1996-1998. Recruitment strategies included targeted mail outs, provision of the program brochure in English and the main languages spoken in the areas and media promotion in ethnic newspapers and on ethnic radio stations. In Victoria, only 55% of the population with diabetes currently access eye care services at the recommended intervals. This program was able to increase compliance with guidelines to 70% among people with diabetes that had not had a recent eye examination. A total of 1,197 people with diabetes were screened for diabetic retinopathy. Of the 1,197 people who were screened, 620 (15% of the estimated number of people with diabetes) had not had their eyes examined in the past two years. This pilot study identified strategies to encourage people with diabetes to have their eyes examined at the recommended intervals. PMID- 11035556 TI - Cataract surgical coverage: results of a population-based survey at Nkhoma, Malawi. AB - AIM: To determine the cataract surgical coverage (CSC) in the close proximity of Nkhoma Hospital, Malawi. METHOD: A rapid epidemiological assessment of the population aged 40 years and over was undertaken to determine the proportion of eyes and patients with cataract blindness that had previously received cataract surgery within a 10-mile radius of the hospital. RESULTS: A total of 993 people aged 40 years and above from all 31 small villages in the 10-mile radius were examined. The prevalence of blindness in this age group was 3.72%, of which 62% was due to cataract. The cataract surgical coverage for people with blinding cataract was calculated at 14.8%, and for eyes with cataract (visual acuity <3/60) 8.5%. CONCLUSION: Nkhoma Hospital has provided affordable cataract surgery for more than 20 years. Despite this, within a ten-mile radius of the hospital, only 1 in 7 people with bilateral blindness from cataract has received surgery. PMID- 11035557 TI - Angle-supported refractive implantation in stable, adult accommodative esotropia. AB - A patient with hyperopia and accommodative esotropia was implanted bilaterally with hyperopic angle-supported refractive implants. Control of the esotropia was achieved as was some improvement in fusional ability. After three years, the endothelium remains healthy, the eye is quiet and control of the esotropia is maintained. PMID- 11035558 TI - Listing's law in strabismus and amblyopia: a preliminary report. AB - We investigated whether Listing's law applies in patients with diminished or no stereopsis due to strabismus or amblyopia. Eye movements of normal subjects and patients with strabismus and/or amblyopia were recorded during monocular and binocular fixation; from these data the shape and relative orientation of displacement planes were calculated. In normal subjects, monocular or binocular fixation did not influence the thickness and relative orientation of displacement planes. No differences were found between normals and the patient with amblyopia due to anisometropia. In one patient with strabismus but without amblyopia, the orientation of displacement planes depended on the fixation conditions; a coupling between horizontal vergence effort and plane orientation was observed. Patients with amblyopia and strabismus showed abnormally shaped and/or abnormally orientated displacement planes, which depended on the fixation conditions. Differences between both eyes in the shape of the planes were also observed. These results show that normal Listing behavior can be present in subjects with diminished stereopsis. They also show that normal stereopsis does not necessarily mean normal Listing behavior, suggesting that Listing's law is mainly a result of motor strategy. PMID- 11035559 TI - Stimulus deprivation myopia in human congenital ptosis: a preliminary report of 50 unilateral cases. AB - OBJECTIVE: To investigate the frequency of hyperopia and suspected stimulus deprivation myopia in human congenital ptosis. METHODS: Ametropia was evaluated in both eyes of 50 patients with congenital ptosis. The age at investigation was one year and older, average 6 years and 10 months. The amount of refraction was documented as the spherical equivalent (100% cycloplegia). Differences between the paired eyes of 1.0 dpt or more were defined as anisometropia. Statistical analysis was performed using the chi-square and sign tests. RESULTS: The frequency of myopia was lower (7/50: 14%) than that of hyperopia (43/50: 86%) in the ptotic eye (p<0.001). However, myopia occurred more often in the ptotic eye (7/50: 14%) than in the fellow eye (3/50: 6%, p>0.3). Myopic anisometropia was found only in the ptotic eye (5/50 vs. 0/50, p = 0.06), but was not more frequent than hyperopic anisometropia: (5/50 vs. 9/50, p>0.4). A covered center of the pupil in 27 out of 50 eyes was not associated significantly with myopia (2 of 27 versus 5 of 23, p>0.2). Altogether, we found a significantly higher rate of myopia <-1 dpt and hyperopia > 2 dpt: 10% vs. 1.4% and 40% vs. 10.2% (p<0.001) in comparison with normal school children. CONCLUSION: In summary, the following three findings were noted, of which the first two were expected and the third was not. 1. Compared with the normal population, there was an overall higher frequency of myopia in unilateral congenital ptosis. 2. There was a higher frequency of myopia in the ptotic than in the fellow eye. 3. Compared with the normal population, there was also a higher frequency of hyperopia. The clinical presentation of human congenital ptosis may be influenced by compensating head posture, strabismus, accommodation and biochemical effects, and the condition may therefore differ from the classical well-defined animal model of stimulus deprivation myopia. PMID- 11035560 TI - Preliminary report: saccade analysis in acute Graves' ophthalmopathy. AB - A prospective study on the saccadic properties of 10 patients with acute Graves' ophthalmopathy (GO) compared to 10 healthy individuals was performed using the induction scleral search coil. No significant changes were observed among six patients with moderate disease. In four patients suffering from optic nerve compression, however, a substantial decrease of saccadic velocity and related parameters was detected. The absence of saccadic changes in earlier moderate stages of GO seems to be based on adaptation of the central saccadic generator. The velocity reduction in connection with a marked elevation of intraorbital pressure could be indicative of paretic motoneuron disturbances. PMID- 11035561 TI - Measurement of venous outflow pressure in the central retinal vein to evaluate intraorbital pressure in Graves' ophthalmopathy: a preliminary report. AB - PURPOSE: To evaluate the intraorbital pressure in patients with Graves' ophthalmopathy (GO) in relation to the intraocular pressure (IOP) and proptosis and to find out whether optic nerve compression is predictable. METHODS: The venous outflow pressure (VOP) in the central retinal vein was measured by the perviously described technique of oculodynamometry.1 Since the central retinal vein passes through the orbit, the VOP cannot be lower than the intraorbital pressure if outflow is to be guaranteed. The IOP was measured either in primary position or with slight chin elevation to avoid restriction of the globe. Fifty seven patients underwent a complete ophthalmologic examination, including VOP measurements, Hertel exophthalmometry and visual fields. RESULTS: The IOP in primary position ranged between 10 and 29 mmHg and in most (n=54) cases the VOP was 0-4 mmHg higher than the IOP. These patients had neither scotomas nor visual deterioration during an observation period of up to 2 years. In those cases (n=3) where the difference between IOP and VOP was 35 mmHg, the patients developed scotomas and visual deterioration and had to be treated (high-dose steroids or orbital decompression). The elevation in VOP did not correlate with the degree of proptosis. In one unilateral case, treatment of high IOP (32 mmHg) with dorzolamide drops led to a decrease in visual acuity of two lines, inferior field depression and relative afferent pupillary defect. The difference between IOP and VOP was 10 mmHg. Stopping treatment normalized visual function, the IOP rose to its original level and the difference between IOP and VOP was 4 mmHg. CONCLUSION: The increased IOP in GO is not caused by primary glaucoma but by elevated intraorbital pressure. The difference between IOP and VOP must be <5 mmHg to guarantee normal perfusion. We interpret these findings to suggest that loss of visual acuity and visual field defects may not only be caused by optic nerve compression at the apex but also by deterioration of optic nerve head perfusion. PMID- 11035562 TI - Vision screening survey of all children starting primary school in 1998 in the Federal State of Saarland, Germany. AB - BACKGROUND: The excellent co-operation with the pediatric public health service of the local health authorities enables us to present the collected results of a vision screening survey of all children starting primary school in 1998 in the Federal State of Saarland. The aim is to analyze the prevalence of amblyopia, strabismus, reduced visual acuity, refractive errors and severe visual impairment in one complete grade of pre-schoolers. METHODS: The examination parameters had been determined in co-operation with the Department of Pediatric Ophthalmology at the University of Saarland and had been fitted to the needs and abilities of lay persons (health workers) doing a vision screening as part of a general health check-up. Parameters were: visual acuity (Rodenstock R21), color vision (Ishihara), stereopsis (Lang), and test for latent hyperopia. Referral to an ophthalmologist if: visual acuity < 0.7; difference in visual acuity in both eyes of more than one line, or any other pathological test result. A total of 12,192 children were screened. RESULTS: The preventive pediatric examinations were complete in 5756 children (56.4%), incomplete in 4449 children (43. 6%) and in 1987 children (16.3%) the degree of completion could not be determined. Eyes: pathological findings in 41.7%. Reduced visual acuity in 30.8%, color vision defects in 1.3%, severe visual impairment in 0.3%. Pathological findings in other organ systems: skeleton 33.5%, teeth 32.6%. For the urban confederacy of Saarbrucken: referrals to ophthalmologists: n=1108. No feedback information: 380; refractive correction: 226; recommendation for regular checks: 346; no pathological findings: 156. CONCLUSION. The high percentage of pathological findings in the vision screening of 12,192 pre-schoolers is an important confirmation that there is a need for a preventive ophthalmologic examination before the age of six. Only an area-wide ophthalmologic vision screening around the second or third year of life can effectively reduce the high prevalence of pathological findings at the time of starting primary school. To improve the present screening situation, networks between ophthalmologists and pediatricians would be beneficial. PMID- 11035563 TI - Preliminary report: examination of young children with Lea symbols. AB - BACKGROUND: Lea symbols are highly sensitive for detection of amblyopia in cooperative patients. They are favorable for visual acuity assessment in childhood. Therefore, we assessed age-related normal values and interocular differences of Lea symbol visual acuity. METHODS/PATIENTS: We reexamined 50 out of 193 children aged 21 months to 7 years who came for a routine pediatric examination between January and November 1999. Lea symbol acuity (Lea Symbols Single Symbol Book (LS) and Lea 15-Line Folding Distance Chart (CLS)) and Landolt C acuity (single (LC) and crowded with 2.6' inter-optotype distance (LC(2.6))) were measured. A three out of four criterion was used. Strabismus and any organic eye disease were excluded by orthoptic and ophthalmologic examination, consisting of biomicroscopy, ophthalmoscopy, retinoscopy or refractometry, cover test or Hirschberg test and Lang Stereotest. RESULTS: Only 26% of the parents (50 out of 193) accepted an examination in our hospital. In 35 (32) of the 50 children, visual acuity could be measured in both eyes separately with single (crowded) Lea symbols, while 26 (25) children could be examined in both eyes monocularly with the Landolt-C with single (crowded) optotypes. Except for one 3-year-old boy, all of the children older than 30 months could be tested with single Lea symbols. Lea acuity surpassed Landolt acuity. The difference was about 1.5 lines (1.5 dB) for both the single and the crowded optotypes. In 63% (69%) of the children who could be tested monocularly, LS acuity (CLS acuity) was higher than 0.8 (0.63). 89% (83%) of the children had an interocular difference of maximum 1 line for single (crowded) Lea symbols. CONCLUSIONS: The youngest child whose visual acuity could be assessed with Lea symbols was 23 months old. Almost every child older than 30 months could be tested with Lea symbols. Lea acuity higher than 1 and an interocular difference less than 2 lines is not suspect for amblyopia. Children with a difference of more than one line should be reexamined. PMID- 11035564 TI - How do the corresponding eyes avoid disturbing contour overlaps? Historical answers (1859 - 1981). AB - The visual system has a limited capacity to fuse overlapping impressions. It takes advantage of three principles of organization: (1) The mixing of different impressions resulting in a new entity (compensation principle); examples are the mixing of colors, dark and light at the retina, Fechner's paradox. The twelve researchers whose work is reported here analyzed two other unifying principles: (2) Selection by successive contrasts (polarization principle) and (3) subordination in a hierarchy of vision (differentiation principle). PMID- 11035565 TI - External quality assessment in the examination of blood films for malarial parasites. PMID- 11035566 TI - Modified triple stain (carbol fuchsin/alcian blue/hematoxylin-eosin) for the identification of Helicobacter pylori. PMID- 11035567 TI - The challenge of evaluating the patient with chest pain. PMID- 11035568 TI - The legacy of Laennec. PMID- 11035569 TI - Neonatal bilirubin testing practices: reports from 312 laboratories enrolled in the Colege of American Pathologists Excel Proficiency Testing Program. AB - OBJECTIVE: To characterize laboratory practices for neonatal bilirubin testing and to identify opportunities for improvement. DESIGN: A voluntary self assessment questionnaire was used to assess the laboratory practices of 312 laboratories subscribing to the College of American Pathologists (CAP) Excel Chemistry Proficiency Testing Program. RESULTS: A range of preanalytic and analytic practices were reported. The most notable problems identified were the use of overly long capillary puncture devices (17.8%), failure to protect the specimen from light prior to analysis (3.4%), failure to use a control containing >171 micromol/L bilirubin (26.1%), use of single-level control (4.2%), and lack of established linearity limits (7.0%). CONCLUSION: Opportunities exist to improve specimen collection, processing, and analysis for some physician-office and small-hospital laboratories. PMID- 11035570 TI - Neuronal hypertrophy in acute appendicitis. AB - OBJECTIVE: The pathogenesis of appendicitis remains poorly understood. However, there is increasing evidence of involvement of the enteric nervous system in immune regulation and in inflammatory responses. This study was set up to characterize the status of the enteric nervous system in normal and in inflamed appendixes. METHODS: S100- and 2',2'-cyclic nucleotide 3' phosphodiesterase positive Schwann cells, synaptophysin, and neuron-specific, enolase-positive nerve fibers and tryptase-positive mast cells were evaluated with immunohistochemical staining in surgically resected appendixes from 20 children with histologically proven acute appendicitis (HA), 10 histologically normal appendixes (HN) from patients with a clinical diagnosis of appendicitis, and 10 normal appendixes from patients undergoing elective abdominal surgery. Immunostained sections were subjected to quantitative image analysis. The number and size of ganglia and the number of nerve fibers, Schwann cells, and mast cells in each tissue compartment was quantitatively or semiquantitatively measured. RESULTS: Increased numbers of fibers, Schwann cells, and enlarged ganglia, widely distributed in the muscularis externa and submucosa, were seen in all HA appendixes and in 4 of 10 HN appendixes. The number and size of ganglia in muscularis externa and in the submucosa of appendixes with HA were significantly greater compared with those in control appendixes (P <.001). A significantly increased number of individually stained nerve fibers and Schwann cells (P <.05) were present in the muscularis externa in HA appendixes compared with control appendixes. Significantly increased numbers of tryptase-positive mast cells (P <.05) were present in the submucosa, muscularis, and especially in the lamina propria in HA specimens, compared with that of control tissue. CONCLUSIONS: The significant increase in neural components and mast cells in acute appendicitis is unlikely to develop during a single acute inflammatory episode. This suggests an underlying chronic abnormality as a secondary reaction to repeated bouts of inflammation, obstruction, or both. These results challenge our current understanding of the pathophysiological processes that give rise to acute appendicitis. PMID- 11035571 TI - Long-term experience with an accelerated protocol for diagnosis of chest pain. AB - CONTEXT: More than 6 million patients present annually with chest pain suggestive of acute coronary syndrome. Rapid and accurate diagnosis is essential for best clinical outcomes, for optimal management of hospital resources, and for minimizing medicolegal exposure. OBJECTIVE: To evaluate the clinical and cost outcomes of an accelerated protocol for chest pain triage in a community-based hospital of moderate size. METHODS: One hundred successive patients with chest pain were diagnosed according to the Traditional Chest Pain Protocol, which included testing of serial blood samples for creatine kinase (CK)-MB and total CK. These patients were also subjected to the Accelerated Chest Pain Protocol under evaluation, which included testing at shortened intervals for myoglobin and cardiac troponin I in addition to CK and CK-MB. Diagnostic sensitivity and specificity were compared versus the final assigned diagnosis. The Accelerated Chest Pain Protocol was implemented for routine use. Follow-up evaluations were conducted at 1 month (test group A, N = 180) and 22 months (test group B, N = 180). Costs for diagnosis and treatment of the 2 test groups were compared with those for the control group. RESULTS: The 2 protocols had equivalent specificity values (99%). The sensitivity of the Accelerated Chest Pain Protocol was higher than that of the Traditional Chest Pain Protocol (95% vs 58%). Cost savings of 29% and a reduction in length of stay of 33% were achieved in test group B versus the control group. CONCLUSIONS: The Accelerated Chest Pain Protocol improved the accuracy and timeliness of diagnosis of acute coronary syndrome while reducing costs. PMID- 11035572 TI - Bone metastases from thyroid carcinoma: a histopathologic study with clinical correlates. AB - CONTEXT: Only limited information exists on the pathologic aspects of thyroid carcinomas with bone metastases, most large studies having concentrated mainly on their clinical features. OBJECTIVE: To study in detail the morphologic features of thyroid carcinomas with skeletal metastases. DESIGN: Seventy-nine cases of thyroid carcinoma with bone metastases treated at Memorial Sloan-Kettering Cancer Center, New York, NY, between 1964 and 1998 were investigated, with emphasis on the pathology of the primary and/or metastatic tumors and comparison of the morphologic features of the tumors at both the sites, wherever possible. The tumors were also compared for various clinical parameters. RESULTS: The cohort consisted of 22 papillary, 17 follicular, 16 insular, 10 anaplastic, 9 Hurthle cell, and 5 medullary carcinomas. Of these cases, 68% had poorly differentiated or undifferentiated features in the primary and/or metastatic tumors. The metastatic tumors were better differentiated than the primary in one third of the cases (6 of 18). Only one case showed a less differentiated metastasis. The overall 5- and 10-year survival probabilities after the bone metastases were 29% and 13%, respectively (Kaplan-Meier method). Although both the tumor type and differentiation seemed to affect survivals after bone metastasis (P =.007 and.012, respectively) (log-rank test), this was primarily due to the much worse prognosis in the cases of anaplastic and medullary carcinoma. Cases of Hurthle cell carcinoma showed the longest median survival. There was no significant difference in survival among patients up to or older than 45 years at the time of metastases (P =.31). CONCLUSIONS: Most thyroid carcinomas with bone metastases are of papillary type, and most have poorly differentiated or undifferentiated features. The influence of the microscopic tumor type and tumor differentiation on survival after bone metastasis primarily appears to be due to the much worse prognosis among anaplastic and medullary carcinomas. Age at diagnosis of bone metastases does not influence survivals. PMID- 11035573 TI - Relationship of myoma cell size and menopausal status in small uterine leiomyomas. AB - CONTEXT: Although myomas shrink after menopause, the cellular mechanism for this phenomenon has received little attention. It was recently demonstrated that fibrous degeneration is significantly associated with postmenopausal status in both small and large myomas. OBJECTIVE: The purpose of the present study was to evaluate whether reduction in myoma cell size is also associated with postmenopausal status in small myomas. DESIGN: Tumor size and patient age have also been related to fibrous degeneration in small (<1 cm) myomas. Therefore, in the present study, 10 pairs of premenopausal and postmenopausal small myomas were matched within 3 years for patient age, within 1 mm for size, and within 1 grade for degree of fibrous degeneration. Most of the women were in their 50s, the decade during which postmenopausal fibrous degeneration in small myomas is most prevalent. Myoma cell size was derived by morphometric evaluation of relative myoma cell area (correcting for percentage of stroma, as measured by point counting) and by direct counting of the number of myoma cells per unit area in trichrome-stained sections. RESULTS: Small myomas from postmenopausal women had significantly (P <.05) smaller cell sizes than did size-matched myomas from age matched premenopausal women. Myoma cell sizes and nucleus-cell (N/C) ratios were highly variable, especially in premenopausal myomas. CONCLUSIONS: Reduction in myoma cell size is significantly associated with postmenopausal status in small uterine leiomyomas and may be an important mechanism for postmenopausal shrinkage of myomas. In addition, the high variability of myoma cell size and N/C ratio may further support the somatic mutation theory (ie, the theory that diverse mutations may account not only for variations in the growth potential of uterine myomas, but also for variations in their cellular details). PMID- 11035574 TI - Incidental prostatic adenocarcinomas and putative premalignant lesions in TURP specimens collected before and after the introduction of prostrate-specific antigen screening. AB - BACKGROUND: Since the introduction of prostate-specific antigen (PSA) screening for the detection of prostatic adenocarcinoma (PCA), there has been an increase in the incidence of stage T1c PCA. The purpose of this study was to compare the frequency of incidental PCA found in transurethral resection of prostate (TURP) specimens for a 14-month period during 1989-1990 (before PSA screening was available) with the incidence of PCA for a 32-month period during 1997-1999 (after PSA screening became available). DESIGN: Consecutive TURP specimens from the 2 time periods were reviewed to identify incidental PCA, prostatic intraepithelial neoplasia (PIN), and atypical adenomatous hyperplasia (AAH). Cases of TURP for palliative treatment of known advanced PCA were excluded from the study. All TURP specimens were fixed in 10% buffered formalin and were processed according to the same protocol. RESULTS: We reviewed 533 and 449 TURP specimens for the time periods 1989-1990 and 1997-1999, respectively. Comparison of the results for these 2 time periods revealed that the combined prevalence of T1a and T1b PCA decreased over time from 12.9% to 8.0% (P =.06) with the introduction of PSA screening. A new group of T1c PCA was established in the post PSA screening period of 1997-1999. There were no statistically significant differences in the incidences of T1a PCA, PIN, and AAH in TURP specimens for the 2 time periods. CONCLUSION: The decreased incidence of T1b PCA in TURP specimens for the 1997-1999 period represents a shift in PCA staging. Some PCAs previously staged as T1b are now staged as T2 carcinomas, as a result of PSA screening and earlier clinical detection. The introduction of PSA screening has had no influence on the incidence of T1a PCA, PIN, or AAH in TURP specimens. PMID- 11035575 TI - Multiparameter immunohistochemical analysis of the cell cycle proteins cyclin D1, Ki-67, p21WAF1, p27KIP1, and p53 in mantle cell lymphoma. AB - BACKGROUND: Mantle cell lymphoma (MCL) is characterized by overexpression of cyclin D1, a G1 cyclin that participates in the control of cell cycle progression at the G1 to S phase transition. In addition to cyclin D1, other cell cycle regulatory molecules may be involved in the proliferation and progression of MCL. Mutation of p53, deletion of p16(INK4a), and loss of p21(WAF1) expression have been reported in some cases of blastoid MCL. OBJECTIVE: We sought to examine levels of expression of these proteins in typical and blastoid MCL and to determine whether differences were present between these subtypes of lymphomas. DESIGN: A retrospective series of typical and blastoid MCLs was evaluated for expression of the cell cycle-related proteins cyclin D1, p21(WAF1), p27(KIP1), Ki 67, and p53, as well as mitotic index. Paraffin-embedded archival tissues from 24 MCL specimens (17 typical, 7 blastoid) were immunostained with antibodies to p21(WAF1), p27(KIP1), p53, Ki-67, and cyclin D1. The percentage of positive cells for each specimen was estimated by counting 1500 cells under oil immersion microscopy. Levels of antigen expression were compared for the typical and blastoid MCLs. The mitotic index was estimated using twenty 100x oil immersion fields (OIFs) for each specimen. RESULTS: Cyclin D1 expression was seen in 22/24 specimens (92%). Blastoid MCLs were characterized by a significantly higher mean mitotic index (>20 mitoses/20 OIFs) and Ki-67 index (>45%) when compared with typical MCLs (P <.001 and P <.008, respectively; Fisher's exact test). High expression of p27(KIP1) (>25% staining) was seen more frequently in typical MCLs than in the blastoid variants (P =.03; Fisher's exact test). No significant differences were found between typical and blastoid MCLs for the expression of p21(WAF1) or p53. CONCLUSIONS: A significantly higher mitotic index and Ki-67 index were found in blastoid MCLs as compared with typical MCLs. Low p27(KIP1) expression was associated with the blastoid MCL variant. These findings confirm the high proliferative nature of blastoid MCL and suggest a role for p27(KIP1) in the negative regulation of the cell cycle in MCL. PMID- 11035576 TI - Hantavirus pulmonary syndrome is distinguishable from acute interstitial pneumonia. AB - CONTEXT: Hantavirus pulmonary syndrome (HPS) and acute interstitial pneumonia (AIP) can share similar clinical presentations. AIP is an acute, diffuse lung disease that has some clinical features suggesting a viral infection, although causative agent(s) have not been identified. OBJECTIVE: To clinically, histologically, and immunohistochemically compare cases of HPS to cases of AIP and to determine if any cases of AIP were actually examples of HPS. DESIGN: Seven cases of HPS and 9 cases of AIP were compared clinically and histologically by semiquantitative grading of features in lung tissue. The cases were also evaluated immunohistochemically for the presence of hantaviral antigens. RESULTS: Hantavirus pulmonary syndrome had a shorter clinical duration and more acute changes histopathologically; AIP was of longer clinical duration and was usually accompanied by histologic evidence of organization. Hantavirus pulmonary syndrome was distinguished by the presence of immature leukocytes in the pulmonary vasculature. No hantaviral antigens were identified immunohistochemically in the 9 case of AIP. Hantaviral antigens were identified in all 7 cases of HPS. CONCLUSION: Cases of AIP and fatal cases of HPS can generally be distinguished on clinical and histologic grounds, and this distinction can be further confirmed immunohistochemically. PMID- 11035577 TI - Estrogen and progesterone receptors in non-small cell lung cancer in 248 consecutive patients who underwent surgical resection. AB - CONTEXT: Different authors have reported estrogen receptor (ER) expression between 0% and 96.8% and progesterone receptor (PR) expression between 21.8% and 34.7%. OBJECTIVE: To examine the discrepancies in the literature regarding the expression of ERs and PRs in non-small cell lung cancer. DESIGN: Retrospective analysis. SETTING: A referral tertiary care center. PATIENTS: We reviewed 248 consecutive cases of stage I and II non-small cell lung cancers. METHODS AND RESULTS: Sections of formalin-fixed and paraffin-embedded tumor tissue were stained with ER and PR monoclonal antibodies using the avidin-biotin complex detection system with antigen retrieval. Men represented 66.1% of the patients, and women represented 33.9%. Large cell (undifferentiated) carcinoma constituted 10.4% of the entire population; squamous cell carcinoma, 39.1%; adenocarcinoma, 33.0%; and bronchoalveolar carcinoma, 17.3%. Patients with stage I disease represented 77.0% of the population. In this patient population, we found no nuclear or cytoplasmic expression of either ERs or PRs (95% confidence interval, 0%-1.2%). CONCLUSIONS: The absence of expression of ERs and PRs differs from previous articles, which use a variety of techniques, impairing a meaningful comparison of data. In addition, the presence of ER and PR expression in a lung carcinoma is supportive of a nonpulmonary primary tumor metastatic to the lung. The absence of their expression in non-small cell lung cancer does not support a role of these transcription factors in initiating and maintaining this neoplastic process. PMID- 11035578 TI - Cellular origin of gastrointestinal stromal tumors: a study of 27 cases. AB - CONTEXT: Interstitial cells of Cajal (ICCs), also known as pacemaker cells, are cells in the gastrointestinal tract that play a role in the control of gut motility. The ICCs express the c-kit proto-oncogene encoding a type III tyrosine kinase (KIT) receptor, a ligand that is known as stem cell factor (SCF). The maturation of ICCs is dependent on SCF-KIT interaction. The cellular origin, differentiation, nomenclature, and prognosis of gastrointestinal stromal tumors (GISTs) are controversial. OBJECTIVE: To test the hypothesis that GISTs originate from CD34-positive stem cells and differentiate toward an ICC phenotype. MATERIALS AND METHODS: We studied 27 cases of smooth muscle differentiated GISTs collected for 14 years (1985-1999), including 8 benign (leiomyoma), 15 malignant primary (leiomyosarcoma), and 4 metastatic to the liver. Immunohistochemical studies of selected lineage-directed monoclonal antibodies of c-kit (CD117), CD34, vimentin, desmin, alpha-actin, S100, and MIB-1 were performed on both normal and tumor tissues. RESULTS: Immunoperoxidase stains of normal gastrointestinal tract showed both c-kit and CD34-positive cells surrounding the Auerbach ganglia plexus in the gastrointestinal tract. Twenty-seven of 27 tumors strongly expressed c-kit. Fourteen of 27 tumors were positive for CD34. Of the malignant GISTs, 14 of 19 were positive for CD34; of the benign tumors, 0 of 8 were positive for CD34. Thus, CD34 was the best indicator of malignant phenotype. CONCLUSION: This is the first description of benign smooth muscle GISTs negative for CD34. The results of this study suggest that GISTs originate from CD34 positive stem cells and differentiate toward pacemaker cell phenotype. The lack of expression of CD34 in the benign GIST may indicate that benign GISTs are composed of more mature ICCs, whereas malignant GISTs are composed of dedifferentiated ICCs that express CD34-positive stem cells. PMID- 11035579 TI - Myospherulosis in renal cell carcinoma. AB - OBJECTIVE: To study retrospectively the identification, characteristics, and localization of myospherulosis in different types of renal cell carcinomas. DESIGN: Twenty-seven consecutive renal cell carcinomas treated by radical nephrectomy in 1 year were studied. All the tumor and nontumor slides were examined for myospherulosis. The demographic data, histological type of renal cell carcinoma, Robson stage, and Fuhrman grades were recorded. RESULTS: Myospherules were found in 10 cases. They were associated with the clear cell type and a higher nuclear grade. The cell type remained the only significant factor when these 2 factors were tested together with the tumor stage by logistic regression. Myospherulosis tended to be found in younger patients but was not associated with the sex or the side of the tumor. They were scattered within tumor cystic spaces or among sheets of tumor cells. Some of the myospherules might arise from histiocytes or even tumor cells. Compared with previous reports of myospherulosis associated with exogenous or endogenous lipid, the myospherules associated with renal cell carcinoma were smaller and more uniform in size. There is no associated fibrosis or foreign body giant cell reaction. CONCLUSION: As far as we know, this is the first report of myospherulosis occurring in malignant tumors in human, and their associated features are different from those previously described for myospherulosis related to exogenous or endogenous lipid. PMID- 11035580 TI - Enterocytozoon bieneusi as a cause of proliferative serositis in simian immunodeficiency virus-infected immunodeficient macaques (Macaca mulatta). AB - CONTEXT: Enterocytozoon bieneusi is the most frequent microsporidian parasite of human patients with acquired immunodeficiency syndrome and is a significant cause of diarrhea and wasting. Recently, this organism has also been recognized as a spontaneous infection of several species of captive macaques. As in humans, E bieneusi frequently causes enteropathy and cholangiohepatitis in immunodeficient simian immunodeficiency virus (SIV)-infected macaques. OBJECTIVE: To examine E bieneusi as an etiologic agent of nonsuppurative proliferative serositis in immunodeficient rhesus macaques (Macaca mulatta). DESIGN: Retrospective analysis of necropsy material obtained from immunodeficient SIV-infected rhesus macaques. RESULTS: Examination of SIV-infected rhesus macaques (n = 225) revealed E bieneusi proliferative serositis in 7 of 16 cases of peritonitis of unknown origin. The organism could be identified by in situ hybridization and polymerase chain reaction in sections of pleura and peritoneum obtained at necropsy. Serositis was always accompanied by moderate-to-severe infection of the alimentary tract, and morphologic evidence suggested dissemination through efferent lymphatics. Colabeling experiments revealed most infected cells to be cytokeratin positive and less frequently positive for the macrophage marker CD68. Sequencing of a 607-base pair segment of the small subunit ribosomal gene revealed 100% identity to sequences obtained from rhesus macaques (Genbank accession AF023245) and human patients (Genbank accession AF024657 and L16868). CONCLUSIONS: These findings indicate that E bieneusi disseminates in immunodeficient macaques and may be a cause of peritonitis in the immunocompromised host. PMID- 11035581 TI - Perceptions of the ethical acceptability of using medical examiner autopsies for research and education: a survey of forensic pathologists. AB - BACKGROUND: Forensic pathologists face difficult moral questions in their practices each day. Consistent ethical and legal guidelines for autopsy tissue use extending beyond usual clinical and legal imperatives have not been developed in this country. OBJECTIVE: To obtain the perceptions of medical examiners regarding the ethical acceptability of autopsy tissue use for research and education. METHOD: A written, self-report questionnaire was developed and piloted by a multidisciplinary team at the University of New Mexico, Albuquerque. All individuals who attended a platform presentation at the National Association of Medical Examiners Annual Meeting in September 1997 were invited to participate. RESULTS: Ninety-one individuals completed the survey (40% of all conference registrants and approximately 75% of presentation attendees). Sixty-three percent of respondents had encountered an ethical dilemma surrounding autopsy tissue use, and one third reported some professional ethics experience. Perspectives varied greatly concerning the ethical acceptability of using autopsy tissues to demonstrate or practice techniques (eg, intubation, brachial plexus dissection) and of fulfilling requests to supply varying kinds and quantities of tissues for research and education. Most respondents indicated that consent by family members was important in tissue use decisions. Respondents agreed on the importance of basic values in education and research, such as integrity, scientific or educational merit, and formal institutional approval of a project. Characteristics of the decedent did not influence decisions to release tissues, except when the individual had died from a mysterious or very rare illness. Attributes of medical examiners, with the exception of sex, also did not consistently predict responses. CONCLUSION: Significant diversity exists in beliefs among medical examiners regarding perceptions of the appropriate use of autopsy tissues for education and research. There is need for further inquiry and dialogue so that enduring policy solutions regarding human tissue use for education and research may be developed. PMID- 11035582 TI - Effect of a new international reference preparation for proteins in human serum (certified reference material 470) on results of the College of American Pathologists Surveys for plasma proteins. AB - OBJECTIVE: To examine the impact of a new international reference preparation for serum proteins on the among-manufacturer variance in the College of American Pathologists Surveys. DATA SOURCE: The results of the Surveys for the years 1993 1998, supplied by the College of American Pathologists. DATA EXTRACTION AND SYNTHESIS: Mean values for manufacturers' assays were compared for each protein in the quality control samples. Results were separated by the reported reference material from which values for calibrators had been transferred. Standard statistical parameters (means, standard deviations, and coefficients of variation) were calculated from the reported means. CONCLUSIONS: Among manufacturer coefficients of variation have dropped significantly for most serum proteins since the introduction of the new reference material. Possible reasons for continued differences are discussed. PMID- 11035583 TI - Actinomyces and actinobacillus actinomycetemcomitans-Actinomyces-associated lymphadenopathy mimicking lymphoma. AB - We present 2 unusual cases of long-standing, extensive reactive lymphadenopathy secondary to Actinomyces infection, 1 of which was also accompanied by Actinobacillus actinomycetemcomitans-Actinomyces complex infection. To our knowledge, histologic features of lymph node involvement by these organisms have not been previously reported in the literature. One patient had extensive cervical, posterior mediastinal, and abdominal lymphadenopathy. The second patient presented with a submandibular mass and cervical lymphadenopathy. Clinical features strongly suggested lymphoma. The histologic examination of the lymph nodes from both patients revealed reactive follicular hyperplasia, marked interfollicular and capsular fibrosis, and multiple interfollicular microabscesses. Characteristic Actinomyces colonies were identified at the center of the microabscesses in deep sections. Cultures were obtained from the lymph nodes of 1 patient, and were positive for A actinomycetemcomitans. Both patients had poor dental hygiene. Lymphadenopathy subsided with antibiotic therapy and appropriate dental care. PMID- 11035584 TI - Primary pulmonary mucosa-associated lymphoid tissue lymphoma with associated fungal ball in a patient with human immunodeficiency virus infection. AB - We describe a case of primary mucosa-associated lymphoid tissue lymphoma of the lung in a 44-year-old man with human immunodeficiency virus. Low-grade pulmonary lymphomas in human immunodeficiency virus-positive patients are rare and are described most commonly in pediatric patients. The gross, histologic, and molecular features of this unusual case are described. PMID- 11035585 TI - Natural killer-cell lymphoma involving the gynecologic tract. AB - Non-Hodgkin lymphomas (NHL) can involve the gynecologic tract, most often as a manifestation of systemic involvement, and most cases reported have been of B cell lineage. We describe 2 women with natural killer (NK)-cell lymphoma involving the gynecologic tract that initially presented with vaginal bleeding. In case 1, the patient had a stage IE nasal-type NK-cell lymphoma involving the cervix. The tumor was composed of medium-sized, irregular lymphoid cells with angioinvasion and necrosis. In case 2, the patient had a stage IV blastoid NK cell lymphoma/leukemia infiltrating all organs in a hysterectomy and bilateral salpingo-oophorectomy specimen. Subsequent biopsy specimens revealed that the bone marrow and lymph nodes were also involved. The neoplasm was composed of small to medium lymphoid cells with fine nuclear chromatin. Case 1 was assessed immunohistochemically and the neoplastic cells were positive for CD3, CD56, and TIA-1. Case 2 was analyzed using both immunohistochemical and flow cytometry methods. The neoplastic cells were positive for cytoplasmic CD3, CD4, CD7, CD43, CD45, and CD56 and were negative for surface CD3. Both cases were negative for Epstein-Barr virus (EBV) ribonucleic acid (RNA) and molecular studies showed no evidence of T-cell receptor gamma chain gene rearrangements. The immunophenotype and absence of T-cell receptor gene rearrangements support NK-cell origin. We report these cases to illustrate that NK-cell lymphomas can involve, and rarely arise in, the gynecologic tract. PMID- 11035586 TI - Inflammatory myofibroblastic tumor (inflammatory fibrosarcoma) of the bone. AB - Although inflammatory myofibroblastic tumors, originally described in the lung, have recently been recognized to occur in various sites, their origin in bone is exceptional. We present a case of inflammatory myofibroblastic tumor, so-called inflammatory fibrosarcoma, of the bone. The tumor occurred in the iliac bone of a 70-year-old woman. Standard radiographs and magnetic resonance imaging revealed a destructive bone tumor that expanded into the soft tissue. Although the patient underwent excision of the hemi-iliac bone, multiple pulmonary metastases were noted 1 year after the operation. On histologic examination, the tumor was found to be composed of a sarcoma-like cellular area and a hypocellular fibrous area. Inflammatory cell infiltration into the tumor was a distinctive feature and is analogous with that of conventional inflammatory myofibroblastic tumor or inflammatory fibrosarcoma of the soft tissue. This is the first report to our knowledge of an inflammatory myofibroblastic tumor of the bone with distant metastasis. PMID- 11035587 TI - Epididymal rhabdomyoma: report of a case, including histologic and immunohistochemical findings. AB - Genital rhabdomyoma is a rare tumor of skeletal muscle origin that is usually found in the vulvar area of young women. The English literature contains only 2 previous case reports involving men, both of whom were 19 years old. One of these lesions originated in the tunica vaginalis of the testis, and the other originated in the prostate gland. We present the clinical, histologic, and immunohistochemical findings of an epididymal rhabdomyoma in a 20-year-old man. To our knowledge, this is the first such case reported in this location. PMID- 11035588 TI - Epstein-Barr virus-associated high-grade B-cell lymphoma of mucosal-associated lymphoid tissue in a 9-year-old Boy. AB - We report an unusual case of Epstein-Barr virus (EBV)-associated mucosal associated lymphoid tissue (MALT) lymphoma involving the lungs, kidneys, and axillary lymph nodes in a child with congenital hypoadrenalism and panhypopituitarism. The patient presented with an aggressive clinical course and histologic evolution. Initial biopsies (1994) of the lung and kidney revealed histologic features of low-grade B-cell MALT lymphoma with lymphoepithelial lesions within the renal tubules and bronchial epithelium. Subsequent biopsies (1996, 1997, and 1999) revealed progressively greater cytologic atypia, polymorphism, and necrosis; an increased mitotic rate; and a preponderance of large cells, indicative of progression from a low-grade to a high-grade MALT lymphoma. Immunophenotyping of the lung and lymph node lesions revealed identical surface marker profiles: cells were CD19(+), CD20(+), immunoglobulin (Ig) G(+), kappa(+), lambda(-), CD5(-), CD10(-), CD23(-), and IgM(-), and also negative for T-cell markers. Genotypic analysis demonstrated the presence of immunoglobulin heavy chain rearrangement and monoclonality of EBV in the lung lesion by Southern blot hybridization and polymerase chain re()action (PCR). The clinicopathologic features suggest that these lesions might represent an immunosupression-related continuum of low-grade to high-grade MALT lymphomas. Infection with EBV may have contributed to this tumor's aggressive clinical and histologic evolution. PMID- 11035589 TI - Granulosa cell tumor of the adult type: a case report and review of the literature of a very rare testicular tumor. AB - We report a case of testicular granulosa cell tumor of the adult type in a 48 year-old man. Microscopically, the tumor consisted of round to ovoid cells with grooved nuclei that were arranged in several patterns, including microfollicular, macrofollicular, insular, trabecular, gyriform, solid, and pseudosarcomatous. These cells demonstrated strong immunopositivity with MIC2 (O13) antibody, vimentin, and smooth muscle actin and focal positivity with cytokeratin. Although this type of sex cord-stromal tumor is relatively common in the ovaries, it is still extremely unusual in the testis, and it probably represents the rarest type of testicular sex cord-stromal tumor. PMID- 11035590 TI - Adenoid cystic carcinoma of the ovary. AB - We describe a case of ovarian adenoid cystic carcinoma (ACC) of the left ovary in a 23-year-old woman. The tumor had the typical cribriform pattern of ACC, lacked any component of surface epithelial carcinoma, and showed myoepithelial differentiation. The features of salivary gland-type tumor seen in this case are unusual and different from those of so-called ACC-like carcinomas of the ovary, which only resemble the salivary gland tumor histologically. PMID- 11035591 TI - Hepatocellular carcinoma and non-Hodgkin lymphoma in a patient with chronic hepatitis C and cirrhosis. AB - Hepatitis C virus (HCV) is a hepatotropic virus, but its genome and replicative intermediates also have been detected in peripheral blood mononuclear cells in patients with chronic hepatitis C. Chronic HCV infection may lead to hepatocellular carcinoma and, in a small percentage of cases, to B-cell non Hodgkin lymphoma. To our knowledge, coexistence of these 2 tumors has not been reported previously. We describe a case of chronic hepatitis C and cirrhosis with 2 small hepatocellular carcinomas and incidental non-Hodgkin lymphoma of a hilar lymph node found during liver transplantation. Although the mechanisms of HCV oncogenesis in hepatocellular carcinoma and in lymphoma are unclear, the presence of these 2 tumors in a single patient are in agreement with the tropism of HCV and its role in oncogenesis. PMID- 11035592 TI - The characteristic appearance of non-alcoholic duct destructive chronic pancreatitis: a report of 2 cases. AB - We report 2 patients with an unusual form of chronic pancreatitis, both of whom were treated for clinical suspicion of pancreatic malignancy. The surgical specimens revealed a dense lymphoplasmacytic infiltration of the main and interlobular branches of the pancreatic duct, causing sclerosis of the duct wall, diffuse irregular lumenal narrowing, extensive parenchymal fibrosis, and organ enlargement. Neither case showed calcifications, fat necrosis, or cyst formation, features usually seen in alcoholic pancreatitis, nor was there any evidence of neoplasia. One patient had an unusual form of acalculous cholecystitis, but without cystic duct inflammation or fibrosis. Both patients recovered well from the surgical procedure and have not had any complications or relapse of their symptoms. To the best of our knowledge, these cases are representative of the recently described non-alcoholic duct destructive chronic pancreatitis, which is thought to be immune-mediated. PMID- 11035593 TI - Concomitant well-differentiated adenocarcinoma and leiomyosarcoma of the uterus. AB - We describe a case of a concomitant well-differentiated endometrial endometrioid adenocarcinoma and leiomyosarcoma of the uterus in a 66-year-old woman who presented with a 6-month history of vaginal bleeding. The patient underwent total hysterectomy for endometrial carcinoma diagnosed by endometrial biopsy. Gross examination of the specimen revealed an endometrial mass bulging into the endometrial cavity and an underlying well-circumscribed nodule separated from the endometrial mass by a myometrial band. Frozen section performed at the time of the total hysterectomy rendered a diagnosis of malignant mixed-mullerian tumor. Histologic examination of the permanent sections revealed well-differentiated endometrial endometrioid adenocarcinoma clearly separated from a high-grade leiomyosarcoma. Differential diagnosis included malignant mixed-mullerian tumor. However, no admixture of carcinomatous and sarcomatous elements was present. There were no heterologous elements. To the best of our knowledge, no similar case has been described in the English literature. PMID- 11035594 TI - Kaposiform hemangioendothelioma of the thymus. AB - Kaposiform hemangioendothelioma is a rare pediatric neoplasm that presents most commonly in the soft tissues. We report the case of a 1-month-old infant who presented with stridor and was found to have a diffusely infiltrating tumor in the thymus that extended into the pericardium and up the carotid sheaths. Histologic examination revealed a vascular tumor infiltrating among the lobules of the lymphocyte-depleted thymus. The lesion had features of both a capillary hemangioma and Kaposi sarcoma. Immunoperoxidase studies on formalin-fixed, paraffin-embedded tissue demonstrated the neoplastic endothelial cells to be positive for vascular markers CD31 and CD34. Antibody to factor VIII-related antigen labeled feeding vessels, but failed to stain the lobules of tumor. Although these tumors have been treated in a fashion similar to capillary hemangiomas in the past, it may be important to differentiate Kaposiform hemangioendotheliomas because of their association with Kasabach-Merritt syndrome and recent success with more aggressive chemotherapy regimens. PMID- 11035595 TI - Neutrophilic thrombophagocytosis. AB - Thrombocytopenia as a complication of B-cell lymphoma usually results from the removal of platelets from the circulation by splenic and hepatic mononuclear phagocytes.1,2 In the present paper, we describe a case of B-cell lymphoma with thrombocytopenia in which the platelets were phagocytosed by the patient's own neutrophilic granulocytes. This finding suggests that neutrophilic granulocytes actively participate in immune platelet clearance. PMID- 11035596 TI - Mixed mucus-secreting and oncocytic carcinoma of the thyroid: pathologic, histochemical, immunohistochemical, and ultrastructural study of a case. AB - We report a carcinoma that is, to the best of our knowledge, the first case of a mixed mucus-secreting and oncocytic carcinoma of the thyroid. We also describe the histochemical, immunohistochemical, and ultrastructural features of this tumor. A 59-year-old man complaining of severe bone pain and weight loss underwent clinical and radiologic investigations. The studies revealed a nodule in the left thyroid lobe that was "cold" by (131)I scintiscan and multiple lytic lesions of the skeleton that showed increased uptake by (99m)Tc-Sestamibi scintiscan. Left hemithyroidectomy was performed and the surgical specimen contained a well-circumscribed nodule of 3 cm in the greatest diameter. Light microscopy showed an oncocytic carcinoma with an area of glandular and papillary proliferation of mucin-producing cells. A double histochemical approach (Alcian blue-periodic acid-Schiff and Alcian blue-high-iron diamine) combined with ultrastructural investigation confirmed the presence of true mucus, ruling out the presence of breakdown products of thyroglobulin. Ultrastructural and immunohistochemical studies, together with clinical findings, excluded a possible metastatic origin of the mucin-producing component. PMID- 11035597 TI - Isolated visceral leishmaniasis presenting as an adrenal cystic mass. AB - A 69-year-old woman presented with a large left retroperitoneal suprarenal mass. Radical resection of the left kidney and the mass revealed a cystic adrenal tumor with a weight of 1500 g. Histologic examination showed that the cyst was composed mostly of partially organized clotted blood. The periphery of the mass consisted of a thin rim of cortical and medullary adrenal tissue with superimposed granulomatous chronic inflammation. The infectious nature of the process was manifested by the scattered intracellular and extracellular Leishmania amastigotes that were found throughout the lesion. The differential diagnosis of cystic adrenal masses and the unusual presentation of visceral leishmaniasis are discussed in this context. PMID- 11035598 TI - Pathologic quiz case: fatal pneumonia in an allogeneic bone marrow transplant recipient. PMID- 11035599 TI - Pathologic quiz case: an unusual salivary gland tumor. PMID- 11035600 TI - Pathologic quiz case: a 33-year-old man with an abdominal mass. PMID- 11035601 TI - Images in pathology. Placenta: 6 - 4 = 2. PMID- 11035602 TI - Images in pathology. The placenta in sickle cell disease. PMID- 11035606 TI - Tumors of the peripheral nervous system PMID- 11035608 TI - An introduction to general pathology PMID- 11035607 TI - Diagnostic cytopathology of the breast PMID- 11035609 TI - Comparison of clinical features of acute HIV-1 infection in patients infected sexually or through injection drug use. The Investigators of the Quebec Primary HIV Infection Study. AB - Acute HIV-1 infection (AHI) may present with a clinical picture that represents a diagnostic challenge. We tested the hypothesis that two different routes of infection, that is, sexual versus parenteral, might be associated with a difference in the clinical features of AHI. A prospective cohort of seroconvertors was established in Montreal in private medical clinics and hospitals from February 1996 to May 1999. The prevalence of the symptomatic presentation was almost overlapping within the two groups of newly infected individuals 69% (42 of 61) for men having sex with men (MSM) and 69% (18 of 26) for injection drug users (IDUs; p =.98). Comparison of all types of symptoms and signs as well as their duration was also similar in both groups. Of particular interest, the site of lymph node enlargement was not different despite the estimated sites of intravenous inoculation. Oral and anal ulcers were more frequently observed in MSM than in IDUs (6 versus 0 and 4 versus 1, respectively). Neither the mean CD4+ count (514.8 and 414.7 cells/mm3; p =.14) nor the mean viral load (4.45 and 4.70 log copies/ml; p =.40) were different between the two groups at the time of the first study visit. Our study results clearly indicate that health care workers can expect similar clinical presentation of AHI in MSM and in IDUs despite the different routes of infection. PMID- 11035610 TI - Early diagnosis of HIV-1-infected infants in Thailand using RNA and DNA PCR assays sensitive to non-B subtypes. AB - OBJECTIVES: To evaluate the sensitivity and specificity of RNA and DNA polymerase chain reaction (PCR) for early diagnosis of perinatal HIV-1 infection and to investigate early viral dynamics in infected infants. DESIGN: A cohort study of 395 non-breastfed infants born to HIV-infected mothers in a randomized clinical trial of short-course antenatal zidovudine. METHODS: Infant venous blood specimens collected at birth, 2 months, and 6 months of age were tested by qualitative DNA and quantitative RNA PCR (Roche Amplicor). To determine sensitivity and specificity of DNA and RNA PCR, results were compared with later DNA PCR results and to antibody results at 18 months. The HIV-1 subtype of the mother's infection was determined by peptide serotyping. RESULTS: In the study, 92% of mothers were infected with subtype E. DNA PCR sensitivity was 38% (20 of 53) at birth, and 100% at 2 months (53 of 53) and 6 months (47 of 47). RNA PCR sensitivity was 47% (25 of 53) at birth and 100% (53 of 53) at 2 months. All samples that tested DNA-positive tested RNA-positive. Specificity was 100% for both DNA and RNA testing at all timepoints. For infected infants, the median viral load of RNA-positive specimens was 407,000 copies/ml (5.6 log10) at birth, 3, 700,000 copies/ml (6.6 log10) at 2 months, and 1,700,000 copies/ml (6.2 log10) at 6 months. Infant RNA levels at 2 and 6 months did not differ by maternal zidovudine exposure, or RNA level at birth. CONCLUSION: This RNA PCR assay performed well for diagnosing perinatal HIV subtype E infection, detecting nearly half of infected infants at birth, and 100% at 2 and 6 months, with 100% specificity. Infected infant viral RNA levels were very high at 2 and 6 months, and were unaffected by maternal zidovudine treatment. PMID- 11035611 TI - Greater diversity of HIV-1 quasispecies in HIV-infected individuals with active tuberculosis. AB - OBJECTIVE: A continual increase in intrapatient HIV-1 heterogeneity is thought to contribute to evasion of host immune response and eventual progression to AIDS. Tuberculosis (TB) is diagnosed both early and late during the course of HIV-1 disease and may increase diversity of HIV-1 quasispecies by activating the HIV-1 immune response and increasing HIV-1 replication. We examined whether HIV-1 heterogeneity is altered in HIV-1-infected individuals with TB. METHODS: Blood samples were obtained from 7 HIV-1-infected patients with active TB (HIV/TB patients) and 9 HIV-1-infected patients (HIV patients) in Kampala, Uganda (CD4 counts of 0-650 cells/microl and HIV loads of 700-750,000 RNA copies/ml). The C2 C3 region of the HIV-1 envelope gene (env) was amplified by nested polymerase chain reaction (PCR) from lysed peripheral blood mononuclear cells (PBMCs) of each patient, and then subject to sequencing, clonal-quasispecies analysis and heteroduplex tracking analysis (HTA). RESULTS: HTA of env DNA fragments showed increased heterogeneity in the HIV/TB individuals compared with the HIV group. Further sequence and HTA analysis on ten individual env clones for each patient showed significantly greater HIV mutation frequencies in HIV/TB patients than in HIV patients. CONCLUSION: An increase in HIV-1 heterogeneity may be associated with a TB-mediated increase in HIV-1 replication. However, a diverse HIV-1 quasispecies population in HIV/TB patients as opposed to tight quasispecies clusters in HIV patients suggests a possible dissemination of lung-derived HIV-1 isolates from the TB-affected organ. PMID- 11035613 TI - Predicting HIV RNA virologic outcome at 52-weeks follow-up in antiretroviral clinical trials. The INCAS and AVANTI Study Groups. AB - OBJECTIVE: To determine the ability of intermediate plasma viral load (pVL) measurements to predict virologic outcome at 52 weeks of follow-up in clinical trials of antiretroviral therapy. METHODS: Individual patient data from three clinical trials (INCAS, AVANTI-2 and AVANTI-3) were combined into a single database. Virologic success was defined to be plasma viral load (pVL) <500 copies/ml at week 52. The sensitivity and specificity of intermediate pVL measurements below the limit of detection, 100, 500, 1000, and 5000 copies/ml to predict virologic success were calculated. RESULTS: The sensitivity, specificity, and positive and negative predictive values of a pVL measurement <1000 copies/ml at week 16 to predict virologic outcome at week 52 were 74%, 74%, 48%, and 90%, respectively, for patients on double therapy. For patients on triple therapy, the sensitivity, specificity, and positive and negative predictive values of a pVL measurement <50 copies/ml at week 16 to predict virologic outcome were 68%, 68%, 80%, and 47%, respectively. CONCLUSIONS: For patients receiving double therapy, a poor virologic result at an intermediate week of follow-up is a strong indicator of virologic failure at 52 weeks whereas intermediate virologic success is no guarantee of success at 1 year. For patients on triple therapy, disappointing intermediate results do not preclude virologic success at 1 year and intermediate successes are more likely to be sustained. PMID- 11035612 TI - Didanosine dosed once daily is equivalent to twice daily dosing for patients on double or triple combination antiretroviral therapy. The AI454-147 Team. AB - OBJECTIVE: To compare the antiviral activity, effect on CD4 cell count, and tolerability of didanosine (ddI) administered once daily and twice daily in HIV-1 infected patients receiving ddI with stavudine or zidovudine, with or without a protease inhibitor. The study was designed to demonstrate that once-daily dosing of ddI was not inferior to twice-daily dosing. DESIGN: Randomized, open-label, multicenter, two-arm study. PATIENTS AND METHODS: 121 HIV-1-infected adults on a stable regimen including ddI (twice daily) during the previous 3 months with a stable viral load <10,000 copies/ml started therapy. Of these, 62 were randomized to switch to a combination that included ddI once daily and 59 to continue with ddI twice daily. The ddI dose was 400 mg/day (250 mg/day if body weight was <60 kg). The primary efficacy analysis compared the time-averaged difference (TAD) between the two treatment regimens in change from baseline log10 plasma HIV-1 RNA levels over 24 weeks of therapy, with an equivalence margin between the two treatment groups of <0.5 log10 copies/ml. RESULTS: At week 24, the mean plasma HIV-1 RNA level had increased by 0.31 and 0.17 log10 copies/ml in the ddI once daily and ddI twice-daily groups, respectively. The time-averaged difference between the two groups in change from baseline plasma HIV-1 RNA levels over 24 weeks was (0.05 log10 copies/ml (95% confidence interval, -0.21 to +0.12 log10 copies/ml), indicating that the antiviral activity of ddI once daily is similar to that of ddI twice daily. After 24 weeks of treatment, changes from baseline in CD4 cell counts were similar in the two groups. Both regimens were generally well tolerated. CONCLUSIONS: Once-daily and twice-daily ddI are equally effective at reducing plasma HIV-1 RNA levels when used in a combination regimen with stavudine or zidovudine, with or without a protease inhibitor. PMID- 11035614 TI - Brief report: effect of antiretroviral agents on T-lymphocyte subset counts in healthy HIV-negative individuals. The Italian Registry on Antiretroviral Postexposure Prophylaxis. AB - To study the effect of antiretroviral agents on T-lymphocyte counts in HIV negative individuals, total counts and CD4+ and CD8+ lymphocyte counts were measured in health care workers (HCW) who had been occupationally exposed to HIV who were untreated (164 HCW, group A), or had received antiretroviral postexposure prophylaxis (PEP). PEP included zidovudine (150 HCW, group B), zidovudine plus lamivudine (48 HCW, group C), or zidovudine, lamivudine, and indinavir (85 HCW, group D), at standard dosage for a mean of 30, 27, and 27 days of treatment, respectively. Lymphocyte values were collected after a mean of 44 days following exposure in group A, 48 days in group B, and 30 days both in groups C and D. Student's t-, nonparametric Mann-Whitney, and Kruskal-Wallis tests were used for statistical analysis. A slight increase in mean CD4 (range, 4.8%-6. 7%) and CD8 (range, 1.4%-9.3%) cells/mm3 was observed in each group. Gender, PEP duration, side effects, and follow-up time did not correlate with responses. Data did not vary using CD4 and CD8 percentages. These findings seem to reject any direct effects of antiretroviral agents, independent of retroviral inhibition, on proliferation and redistribution of T lymphocytes, as well as the hypothesized braking of lymphocyte apoptosis. The observed variations could reflect biologic variability. PMID- 11035615 TI - Chemotherapy consisting of doxorubicin, bleomycin, vinblastine, and dacarbazine with granulocyte-colony-stimulating factor in HIV-infected patients with newly diagnosed Hodgkin's disease: a prospective, multi-institutional AIDS clinical trials group study (ACTG 149). AB - To ascertain the results of standard ABVD chemotherapy (doxorubicin, bleomycin, vinblastine, dacarbazine) in HIV-infected patients with newly diagnosed Hodgkin's disease (HD), a nonrandomized, prospective, multiinstitutional clinical trial was conducted by the AIDS Clinical Trials Group (ACTG), in HIV-infected patients with Hodgkin's disease. All patients received the standard ABVD regimen, with granulocyte-colony-stimulating factor (G-CSF). Antiretroviral therapy was not used. Between May, 1992 and August, 1996, 21 patients were added to the study and treated. The median CD4 count was 113 cells/mm3, and 29% had a history of a clinical AIDS-defining condition before diagnosis of HD. Systemic "B" symptoms were present in 90% at diagnosis. Stage IV HD was present in 67%, with bone marrow involvement in 12 (57%). Nodular sclerosis HD was present in 38%, with mixed cellular disease in 31%. Among all patients entered and treated, complete remission (CR) was attained in 9 (43%; 90% confidence interval [CI], 24%-63%), whereas partial response occurred in 4 (19%), leading to an overall objective response rate of 62% (90% CI, 42%-79%). Despite routine use of G-CSF, 10 patients (47.6%) experienced life-threatening neutropenia, with absolute neutrophil counts <500 cells/mm3. In all, nine opportunistic infections occurred in 6 patients (29%) during the study or shortly thereafter. Median survival was 1.5 years. Results of this study suggest that alternative treatment strategies should be explored, including use of chemotherapy together with antiretroviral therapy. PMID- 11035616 TI - Readiness for HIV vaccine trials: changes in willingness and knowledge among high risk populations in the HIV network for prevention trials. The HIVNET Vaccine Preparedness Study Protocol Team. AB - Longitudinal data were analyzed to determine changes in willingness to participate in HIV vaccine efficacy trials and knowledge of vaccine trial concepts among populations at high risk of HIV-1 infection. Gay men (MSM), male and female injection drug users, and non-injecting women at heterosexual risk were recruited (n = 4892). Follow-up visits occurred every 6 months up to 18 months. Willingness was significantly lower at follow-up visits compared with at baseline. Knowledge levels increased for all study populations. Problematic concepts were possible effects of the vaccine on the immune system and lack of knowledge about efficacy of a vaccine before the start of a trial. For concepts concerning safety, blinding, and guarantees of future participation in trials, MSM men had significant increases in knowledge, but little to no change occurred for the other populations. An increase in knowledge was associated with becoming not willing, particularly among MSM with low knowledge levels. At least half of high-risk participants were consistently willing to participate in future vaccine efficacy trials and with basic vaccine education, knowledge levels increased. Continued educational efforts at the community and individual level are needed to address certain vaccine trial concepts and to increase knowledge levels in all potential study populations. PMID- 11035617 TI - Trends in HIV-1 incidence in a cohort of prostitutes in Kenya: implications for HIV-1 vaccine efficacy trials. AB - BACKGROUND: Accurate predictions of HIV-1 incidence in potential study populations are essential for designing HIV-1 vaccine efficacy trials. Little information is available on the estimated incidence of HIV-1 in such populations, especially information on incidence over time and incidence while participating in risk-reduction programs. OBJECTIVES: To examine time trends in HIV-1 incidence in a vaccine preparedness cohort. DESIGN: Prospective cohort study of female prostitutes in Mombasa, Kenya. METHODS: HIV-1 incidence was determined using open and closed cohort designs. Generalized estimating equations were used to model HIV-1 and sexually transmitted disease (STD) incidence and sexual risk behaviors over time. RESULTS: When analyzed as a closed cohort, HIV-1 incidence declined 10 fold during 3 years of follow-up (from 17.4 to 1.7 cases/100 person-years; p <.001). More than 50% of the cases of HIV-1 occurred during the first 6 months after enrollment, and 73% during the first 12 months. When analyzed as an open cohort, HIV-1 incidence density fell during the first 4 calendar years, influenced by accumulation of lower risk participants and variations in study recruitment. Significant declines occurred in both STD incidence and high-risk sexual behaviors during follow-up. CONCLUSIONS: This study documents a dramatic decline in the risk of HIV-1 infection while participating in a prospective cohort, with most seroconversions occurring within 1 year of enrollment. Variations in HIV-1 incidence within high-risk populations should be anticipated during the design of vaccine trials. PMID- 11035618 TI - Expansion of rare CD8+ CD28- CD11b- T cells with impaired effector functions in HIV-1-infected patients. AB - The decline in the number of CD4+ T cells in HIV-1-infected patients is known to be related to the increased number of CD8+CD28- T cells. In this paper, we show that CD8+CD28- T cells from HIV-positive patients have an impaired capability to interact with human endothelial cells. This is due to the dramatic expansion, within this subset, of rare CD11b- cells lacking cell-cell adhesion functions. In 50 HIV-positive patients, 19.5% +/- 6.5% of all T cells were CD8+CD28-CD11b-, whereas only 0.8% +/- 0.4% of all T cells from healthy donors showed this uncommon phenotype. The percentage of circulating CD8+CD28-CD11b- T cells was strongly related to the percentage of CD4+ T cells (r = -0.82). This population is peculiar in terms of HIV infection and was found to possess some characteristics associated with effector functions but its cytotoxic properties were impaired. The percentage of target cells lysed by CD8+CD28-CD11b- was significantly lower than that of cells lysed by its CD11b- counterpart (p <.05) both at low (5:1) or at relatively high (20:1) effector/target ratios. CD8+CD28 CD11b- T cells, which lack the ability to interact with endothelial cells, are likely to accumulate and persist in circulation. The biologic properties of CD8+CD28-CD11b- T cells suggest that these cells might be endstage or aberrant differentiated effector cells. Lack of cell-cell adhesion and impaired cytolytic functions favor the hypothesis of a role for CD8+CD28-CD11b- T cells in the development of immunodeficiency. PMID- 11035619 TI - Are there gender differences in starting protease inhibitors, HAART, and disease progression despite equal access to care? AB - OBJECTIVES: To describe gender differences in starting and response to treatment regimens and long-term clinical outcome in a well-characterized regional population from the Southern Alberta HIV Clinic (SAC) of 1403 patients, where all medical care for HIV, including physician fees, laboratory tests, and antiretroviral drug costs is provided free of charge. DESIGN: Observational cohort study. METHODS: Cox proportional hazards models were used to examine the relative risk of starting treatment regimens and disease progression (new AIDS defining illness or death). RESULTS: There are 126 women in the SAC (9.0%). The median CD4 lymphocyte count at first visit among all patients was 350 cells/mm3, and was significantly higher among women than men (428 cells/mm3 versus 345 cells/mm3, respectively; p = 0.0024). Participating women were less well educated than participating men; 29% of women did not proceed beyond a tenth grade education compared with 13% of men; only 28% of women went to college or received a degree in contrast to 40% of men (p <. 001). The proportion of women in the cohort has increased over the past 5 years (p <.001). During a median follow-up period of 35 months that dates back as far as 1985, 572 patients (40.8%) died or progressed to a new AIDS-defining illness, of whom 30 were women (5. 2%). In a multivariate Cox model stratified by calendar quartile of first visit and adjusted for latest CD4, AIDS status, age, exposure group, education, and prior treatment, women were significantly less likely to start highly active antiretroviral therapy (HAART; defined as at least three antiretrovirals taken consecutively; relative hazard [RH], 0.69; 95% confidence interval [CI], 0.49 0.98; p =.033), significantly less likely to start a protease inhibitor containing treatment regimen (RH, 0.71; 95% CI, 0.52-0.98; p =.040) and significantly less likely to start a HAART regimen including a protease inhibitor (RH, 0.69; 95% CI, 0.48-1.00; p =.049). After adjustment for potentially confounding variables such as CD4 lymphocyte count and treatment regimen, no difference in disease progression was found between men and women (RH, 0.77; 95% CI, 0. 49-1.19; p =.24). Among patients who started HAART, the CD4 lymphocyte count and viral load at starting treatment regimens was similar between men and women, as were the immunologic and virologic response following initiation of treatment. CONCLUSIONS: Despite free access to antiretrovirals, women in the SAC were significantly less likely to start HAART treatment regimens, and the reasons for this need further investigation. Response to treatment was similar between genders. No evidence was found for a poorer long-term clinical outcome in women, but given the proven large clinical benefits of HAART, this may change in the future. PMID- 11035620 TI - HTLV-II and bacterial infections among injection drug users. AB - OBJECTIVE: To examine whether select bacterial infections are associated with HTLV-II infection among injection drug users, we conducted a nested case control study within an ongoing cohort study. METHOD: HTLV-II status was determined by enzyme-linked immunosorbent assay, immunofluorescent assay, and immunoblot. Diagnosis of bacterial pneumonia, infective endocarditis, and skin abscess was confirmed by standardized chart reviews. Three sets of cases were identified based on diagnosis of bacterial pneumonia, infective endocarditis validated by chart review, or self-reported skin abscess. Each case was matched to a minimum of 5 controls by age, HIV status, and study follow-up duration. Risk factors for each bacterial infection were analyzed separately by conditional logistic regression methods. RESULTS: Prevalence of HTLV-II infection ranged from 7% to 11% in cases and controls. The bivariate association of HTLV-II and bacterial pneumonia revealed an odds ratio (OR) of 1.1 (95% confidence interval [CI], 0.6 2.0); the association of infective endocarditis and HTLV-II revealed an OR of 1.7 (95% CI, 0. 7-3.9); and the association between HTLV-II and skin abscess revealed an OR of 1.3 (95% CI, 0.6-2.0). These ORs were unaltered by adjustment for other factors. CONCLUSION: Our results suggest that these three bacterial infections were not significantly associated with HTLV-II infection within a population of injection drug users. Additional associations between HTLV-II infection and disease outcomes merit further exploration. PMID- 11035621 TI - Potent activity but limited tolerance of ritonavir plus indinavir in salvage interventions. PMID- 11035623 TI - Research involves small steps not giant leaps. PMID- 11035622 TI - Drugs for prevention and treatment of postmenopausal osteoporosis. PMID- 11035624 TI - Issues in effective pain control. 1: Assessment and education. AB - Pain is a complex phenomenon to which an individual's response is determined by the interactions of physical, psychological, cultural and social factors (Melzack and Wall, 1988; Woodward, 1995; Horn and Munafo, 1997). It cannot be measured, and self-assessment by patients is increasingly recognized as the most accurate means of evaluating their pain. However, research indicates that even self assessment remains problematic because nurses frequently either do not use pain tools effectively, or do not always accept the self-assessment. Reasons for this seem to be rooted in the attitudes and beliefs of nurses, and inadequate communication. The latter problem could be improved by more effective use of pain tools, but attitudinal problems are more difficult to address once the barriers have become established. This article suggests that the most effective way to prevent such barriers arising is to ensure that nurses receive a focused, substantive input on pain assessment and pain control within their preregistration studies. Referring to proposals put forward several years ago by the Royal College of Nursing (Davis and Seers, 1991), the authors suggest that the process should begin very early in preregistration studies, preferably within the first 6 months, and involve teaching and learning strategies that encourage personal exploration and so are conducive to personal growth and development. In this way, postregistration studies could realistically focus on specialist aspects of pain care more appropriate to 'continuing education'. PMID- 11035625 TI - Ethical arguments for providing palliative care to non-cancer patients. AB - Palliative care professionals have begun to address the issues surrounding the provision of palliative care to non-cancer patients. Yet the situation remains inconsistent and morally unjustifiable. The duty to provide care, non maleficence, beneficence, protecting the patient's best interests and respecting patient autonomy are key responsibilities which palliative care professionals have for all their patients, regardless of their diagnosis. On the grounds of justice as fairness, equality and equity, the current inconsistencies in the provision of palliative care to non-cancer patients are unfair unequal and inequitable. Professionals can no longer ignore their moral responsibility to address these issues and change their practice to include the provision of care for dying patients regardless of their diagnoses. PMID- 11035626 TI - A longitudinal study on dying in a Norwegian hospital. AB - A research project conducted at a Norwegian hospital in 1977 (n = 213 deaths) was repeated in 1987 and 1997 (n = 100 deaths each year). The purpose was to discover if and how terminal care had changed during these 20 years. Data sources included case records, nurses' reports and interviews with close relatives approximately 12 weeks after death. The average stay in hospital decreased by one-third from 1977 to 1997. Active life-prolonging treatment, as defined by use of antibiotics, resuscitation and parenteral fluids, was greatly reduced from 1977 to 1987, but there was an increase in the use of such treatment in 1997. Interestingly, palliative treatments--either broadly defined as the giving of analgesics in general, or more narrowly as the giving of opiates only--have increased steadily over the years and are now offered to most patients. However, the findings demonstrated that the next-of-kin were less well informed about the patient's impending death in 1997 than in 1977. PMID- 11035627 TI - Palliative care nursing in rural Western Australia. AB - This article presents the findings of a study of rural palliative care nurses in Western Australia. The number of rural centres in Western Australia offering palliative care services is increasing; however, at present there is little empirical data available about the roles of the nurses involved. This study was undertaken to begin to correct this deficit. The study examined basic social processes associated with the role of rural palliative care nurses, and identifies issues that affect the nurses' professional practice. A modified grounded theory approach was used to form a conceptual framework that describes rural palliative care nursing. Theoretical sampling techniques were used to identify the six palliative care nurses working in rural Western Australia who participated in the study. Data were generated using in-depth interview and participant observation techniques. Constant comparative analysis of the data was employed to allow concepts to emerge from the data. The central theme that developed is the all-consuming nature of the rural palliative care nurse's role. Three subthemes relating to multiple roles, expectations of nurses, and coping strategies are also discussed. This research explored issues that rural palliative care nurses feel are relevant to their professional practice, and it describes the basic social processes inherent in the rural palliative care nurse's role. Recommendations for nursing research, education, administration and clinical practice are presented. PMID- 11035628 TI - Patients with non-malignant disease deserve an equitable service. PMID- 11035629 TI - A hope is not a promise: fostering hope within palliative care. AB - In palliative care, where patients have to live with a degree of uncertainty as to the possible course their illness may take and the length of life they may expect, hope is a major issue. It has been said that individuals who know they are going to die soon, can be literally dying for hope. Ahmedzai (1994) points out that a major challenge for palliative care is to find ways of inspiring patients towards new directions for hope. This reflective article will explore the nature of hope and discuss the potential problems surrounding false and unrealistic hope, and will focus on how nurses may foster and support hope throughout life-threatening illness and during the palliative phase of life. PMID- 11035630 TI - A new information resource for palliative care professionals. PMID- 11035631 TI - Paediatric palliative care comes of age. PMID- 11035632 TI - A genome scan in families from Australia and New Zealand confirms the presence of a maternal susceptibility locus for pre-eclampsia, on chromosome 2. AB - Epidemiological studies have shown that genetic factors contribute to the etiology of the common and serious pregnancy-specific disorder pre-eclampsia (PE)/eclampsia (E). Candidate-gene studies have provided evidence (albeit controversial) of linkage to several genes, including angiotensinogen on 1q42-43 and eNOS on 7q36. A recent medium-density genome scan in Icelandic families identified significant linkage to D2S286 (at 94.05 cM) on chromosome 2p12 and suggestive linkage to D2S321 (at 157.5 cM) on chromosome 2q23. In the present article, the authors report the results of a medium-density genome scan in 34 families, representing 121 affected women, from Australia and New Zealand. Multipoint nonparametric linkage analysis, using the GENEHUNTER-PLUS program, showed suggestive evidence of linkage to chromosome 2 (LOD=2.58), at 144.7 cM, between D2S112 and D2S151, and to chromosome 11q23-24, between D11S925 and D11S4151 (LOD=2.02 at 121.3 cM). Given the limited precision of estimates of the map location of disease-predisposing loci for complex traits, the present finding on chromosome 2 is consistent with the finding from the Icelandic study, and it may represent evidence of the same locus segregating in the population from Australia and New Zealand. The authors propose that the PE/E-linked locus on chromosome 2p should be designated the "PREG1" (pre-eclampsia, eclampsia gene 1) locus. PMID- 11035634 TI - Teeth and their sex-dependent dimorphic shape in three species of Costa Rican plethodontid salamanders (Amphibia: Urodela). AB - The shape of the teeth and their sex-dependent dimorphic expression in three species of Costa Rican plethodontids (Bolitoglossa subpalmata, Oedipina uniformis and Nototriton abscondens) were studied using light microscopy and scanning electron microscopy. The teeth of the vomerine tooth patches are about one third larger than the teeth of the jaws in B. subpalmata and O. uniformis, whereas all teeth of N. abscondens are of about uniform size. The occurrence of bicuspid tooth germs in the fetus proves that primary teeth are bicuspid in these directly developing plethodontids. Females possess only bicuspid teeth consisting of a pedicel and a crown, as is considered characteristic for urodeles after metamorphosis. Adult males possess conical monocuspid teeth on the premaxillary. These teeth--which are similar to the typical late larval tooth of salamanders presenting a larval stage--are about twice as big as the neighbouring bicuspid maxillary teeth. N. abscondens males possess some monocuspid teeth and teeth of aberrant shapes on the premaxillary and the maxillaries. A tendency to build more monocuspid teeth in the premaxillary region than in the maxillary region can be observed in this species. We suppose that different degrees of sensitivity to androgens in each section of the dental lamina of the upper jaw cause the secondary occurrence of conical monocuspid teeth predominantly on the premaxillary section. PMID- 11035633 TI - A locus for autosomal dominant colobomatous microphthalmia maps to chromosome 15q12-q15. AB - Congenital microphthalmia is a common developmental ocular disorder characterized by shortened axial length. Isolated microphthalmia is clinically and genetically heterogeneous and may be inherited in an autosomal dominant, autosomal recessive, or X-linked manner. Here, we studied a five-generation family of Sephardic Jewish origin that included 38 members, of whom 7 have either unilateral or bilateral microphthalmia of variable severity inherited as an autosomal dominant trait with incomplete penetrance. After exclusion of several candidate loci, we performed a genome-scan study and demonstrated linkage to chromosome 15q12-q15. Positive LOD scores were obtained with a maximum at the D15S1007 locus (maximum LOD score 3.77, at recombination fraction 0.00). Haplotype analyses supported the location of the disease-causing gene in a 13.8-cM interval between loci D15S1002 and D15S1040. PMID- 11035635 TI - The odontoclasts of Ambystoma mexicanum. AB - The resorption of teeth in Ambystoma mexicanum during postembryonal ontogenesis and induced metamorphosis occurs by means of light-microscopic detectable giant cells. These have morphological and functional characters similar to those of odontoclasts of other vertebrates. The multinucleated odontoclasts resorb not only the pedicel (base), but the stalk of the tooth, too. When active, the cells form a ruffled border and a sealing zone. In this way they are able to demineralize the hard tissues of teeth (dentin and mineral of the pedicel) and to dissolve the extracellular matrix. Resorption of enamel has not been observed. Marks of resorption resemble the Howship's lacunae of other tetrapods. TRAP as a typical enzyme of odontoclasts could not be detected histochemically. Dependence of PTH, which is supposed to be necessary for the formation and activation of odontoclasts as well as of thyroxine can be excluded, although the resorbing cells are functionally and cytologically identical with those of other vertebrates. This demands some other mechanism for the formation and regulation of the odontoclasts in A. mexicanum. PMID- 11035636 TI - Tyrosine hydroxylase-like immunoreactive synaptic boutons in the inferior colliculus of the cat. AB - The data on the distribution of catecholaminergic cells and fibers in such a significant subcortical relay auditory center as the inferior colliculus (IC) are both few and controversial, and ultrastructural data are lacking. Young adult mongrel cats of both sexes were used. Following routine preparation procedures, the ultrathin sections were prepared for the ultrastructural examination of tyrosine hydroxylase (TH)-like immunoreactivity. TH-positive neuronal perikarya were not detected in the IC. On the other hand, an appreciable number of TH immunoreactive unmyelinated axons and synaptic boutons were found in all subdivisions of the IC, most often in the nucleus externus, followed by the nucleus pericentralis, and a few were seen in the dorsomedial part of the central nucleus. The boutons measured 0.5-1.8 microns, contained pleomorphic synaptic vesicles, and established symmetrical synaptic contacts almost exclusively with dendrites of small caliber. PMID- 11035637 TI - Ultrastructural changes in human gingival fibroblasts in vitro after exposure to vapour phase smoke components. AB - Tobacco and some of its volatile and non-volatile components have been found to affect many types of cells including gingival fibroblasts. Because normal gingival fibroblast functioning is fundamental to the maintenance of the oral connective tissue as well as to wound healing, we examined the effect of two vapour phase smoke components (acrolein and acetaldehyde) on proliferation and ultrastructure of human gingival fibroblasts (HGFs) in culture. A human gingival fibroblast strain derived from healthy individuals was used in this study. The cells were incubated in the presence of different concentrations of acrolein and acetaldehyde and cell proliferation and fine morphology were evaluated. The results show that acrolein and acetaldehyde produced dose dependent inhibition of HGF viability and alteration of cytoplasmic organelles. The main ultrastructural finding for the HGF cytoplasm was the presence of vacuoles and lysosomal structures which became prominent with increasing concentration of acrolein and acetaldehyde. Our results suggest that the ultrastructural alterations we observed in HGFs may be due to the uptake and storage of acrolein and acetaldehyde by the cells. PMID- 11035638 TI - Microvascularisation of the raphe buccalis in edentulous subjects, fetus, new born and adult. AB - The aim of this study is to describe the microvascularisation of the raphe buccalis located at the inner surface of the cheek. The raphe buccalis is the zone where the maxillary and mandibulary prominences fuse in the embryo during the second month of life. This study was conducted using heads from edentulous subjects, fetuses, newborns and adults, injected with Indian ink in agar, then dissected or sectioned in the three basic planes. The vascular networks of the raphe buccalis are as followed: a deep reticular network, a superficial reticular network and a papillary network. The microvascularisation of the raphe buccalis classifies it as a continuation of the commissure of the mouth and of the mucosa of the cheek. PMID- 11035639 TI - Correlative changes during early morphogenesis of the sacroiliac complex in squamate reptiles. AB - We tested the "limb bud" hypothesis, which explains morphogenetic mechanisms of the formation of the sacroiliac skeletal complex in tetrapods. The hypothesis assumes that: 1) the destruction of the embryonal sacral myomeres and the appearance in their place of a sacral gap filled in with mesenchymal cells favor the development of the sacroiliac complex; and 2) the destruction of myomeres takes place under the influence of limb buds. We studied serial hystological sections of embryos from squamate reptiles with large limb buds (sand lizard, Lacerta agilis L.), small and short-living limb buds (slow worm, Anguis fragilis L.) and without limb buds (adder, Vipera berus (L.)). In embryos of the sand lizard, the hypaxial part of the second sacral myomere degenerated, whereas that of the first one survived in its cranial part. Thus, a large sacral gap was formed where two sacral ribs expanded later. They stretched in a manner similar to the sacral gap across the longitudinal axis of the body, the large ilium lying opposite them. In embryos of the slow worm, the sacral gap was of reduced size and was significantly beveled caudally. The only sacral rib and the upper part of the ilium, which lie within the sacral gap, were beveled in the same manner. In embryos of the adder, myomere destruction was not observed, and sacral ribs and the pelvic girdle did not arise. The obtained results generally agree with the limb bud hypothesis; therefore, it can be said that tetrapods possess a simple and effective morphogenetic mechanism by which the hind limbs create their own support on the axial skeleton. PMID- 11035640 TI - [Scanning electron microscopic findings concerning the formation of the organ of Corti near the helicotrema in guinea pigs]. AB - We report on findings in guinea pigs with objectively tested normal hearing ability. At the apical end of the Corti organ only in the inner row of hair cells the stereociliae are detectable by scanning electron microscopy. Here the row is interrupted several times. Near the helicotrema we find first the inner row of the outer hair cells, then the middle row and finally the outer row. At the beginning of the D-turn their arrangement is disordered. Normally, at the basal end of the D-turn all rows of inner and outer hair cells show their typical formation. This finding should be taken into consideration when making a cochleogram for proving of experimental noise damages. PMID- 11035641 TI - Functional angioarchitectural comparison of the fungiform papillae of rat, rabbit, cat in scanning electron microscopic specimens. AB - The aim of this study was to compare the functional angioarchitectural fungiform papillae (FuP) of three kinds of mammalia, rat, rabbit and cat, with microvascular cast specimens (MVCS) utilizing the corrosive resin cast method in scanning electron microscopy (SEM). FuP distributed geometrically on the dorsal surface of the tongues of the three mammals with different eating habits were of various kinds of form based on their functional and microvascular structure. The basic form of the capillary loop of FuP in the three mammals consisted of the ascending and descending branches of the capillary bed. The basic form of the loop microstructure in rat FuP was a cylindrical or bamboo-basket-like loop structure. In the rabbit, FuPs consisted of a carnation-like structure and in the cat FuP consisted of a half oval fishnet ball-like structure. It can be said conclusively that from this detailed investigation of the intracapillary microvascular architecture of the three mammals, FuP are effective as a sensory organ receiving the taste of foods and liquids by the expansion of the surface area in various functional and microvascular formations in the rat (cylindrical or bamboo-basket-like loop structure), rabbit (carnation-like) and cat (half oval fishnet-ball-like) respectively. PMID- 11035642 TI - Lesions do not provoke GFAP-expression in the GFAP-immunonegative areas of the teleost brain. AB - In the mammalian and avian brains the predominant astroglial elements are astrocytes, and the distribution of GFAP-immunopositivity is rather uneven, some large brain areas being almost devoid of GFAP-immunopositivity. In these areas however, an intense GFAP-immunopositivity appears following injury. In the teleost brain most of the areas are GFAP-immunopositive and ependymoglia predominates. However, a large area, the layer of the sensory neurons in the vagal lobe, is devoid of GFAP. The question arises, whether the lack of GFAP immunopositivity in the teleost brain is also due to the repression of the GFAP production, as in birds and mammals, or due to the lack of cells capable of expressing GFAP. To answer this question, stab wounds were made in the vagal lobe of goldfish, as well as in the tectum, in which moderately dense but highly organized GFAP-immunopositive glia has been detected in intact animals. In the layer of the sensory neurons in the vagal lobe no GFAP-immunopositivity appeared even after lesions had been introduced. In the tectum, a rather slight increase of the intensity of the immunostaining was observed in the glial fibers near the lesions but no typical reactive glia similar to that found in mammals or birds, was observed. The results suggest that a lesion does not provoke GFAP-expression in GFAP-immunonegative brain areas in teleosts, in contrast to what is observed in mammals and birds. PMID- 11035643 TI - Calcitonin gene-related peptide (CGRP) expression in the human neonatal paravertebral ganglia. AB - The distribution of calcitonin gene-related peptide-immunoreactivity (CGRP-IR) in human neonatal paravertebral ganglia was demonstrated by the method of indirect immunohistochemistry. A marked population (up to 21%) of CGRP-IR neurons and varicose nerve fibres was observed. The number of calcionin gene-related peptide immunoreactive neurons varied from ganglion to ganglion in the sympathetic trunk. In addition to its cotransmitter functions, the existence of CGRP in neonatal ganglionic nerve cells was suggested by its inductive and trophic actions on the growth and differentiation of neurons. PMID- 11035644 TI - Ultrastructural identification of specialized endocytic compartments in macrophages of the thymic cortico-medullary zone and germinal centers of peripheral lymphatic organs of the rat. AB - In this study transmission electron microscopy was used to investigate the ultrastructural features of macrophages which are strategically positioned in the thymic cortico-medullary zone and the dark zone of germinal centers of peripheral lymphatic organs in adult Wistar rats. We show that this, morphologically distinct, type of macrophage displays the entire range of cytoplasmic inclusions, which structurally closely correspond to those of endosomal/MHC-II-enriched compartments of antigen presenting cells. The macrophages of the cortico medullary zone and germinal centers contain numerous multivesicular bodies, as well as various kinds of cytoplasmic inclusions ranging from single to aggregated multivesicular/multilamellar bodies to large vacuoles. These multilamellar inclusions are composed of elongated, irregularly shaped cisternae, with abundance of internal membrane profiles and dense bodies. Often, cortico medullary zone and germinal center macrophages contain the typical multilamellar bodies. Polysaccharides are detected by the thiocarbohydrazide-silver proteinate method within the dense bodies of these macrophages. The functional significance of cortico-medullary zone and germinal center macrophages is briefly discussed. PMID- 11035645 TI - Computerized planimetry of normal and abnormal postmortem pancreatograms. AB - The aim of this study was to evaluate postmortem pancreatograms by means of computerized planimetry of the ductal drainage area. This method was applied to a total of 136 pancreatograms from autopsy specimens of the human duodenopancreas, with and without pathological changes. The mean value of the total ductal drainage surface area was 5,054 mm2 for normal specimens and 3,938 mm2 in cases with chronic pancreatitis; this difference was statistically highly significant. The analyses also included measuring the accessory duct drainage area (if an accessory duct was present), and the percentage of its share in the total gland area. This share was significantly larger in cases with chronic pancreatitis (24.4%) in comparison to the normal specimens (16.8%). In conclusion, computerized planimetry of pancreatograms supplies valuable data not only on the pathogenesis of chronic inflammation of the gland, but also on some of the possible predisposing factors for this condition. PMID- 11035646 TI - Anatomical variations of the human suprarenal arteries. AB - This is on anatomical study of the suprarenal arteries and their variations in 30 cadavers aimed at providing in a subsequent article the anatomical basis of arterial segments of the gland. The suprarenal glands were supplied by 3 main groups of suprarenal arteries: superior, middle and inferior. Only the superior and the inferior groups were present in all cases, since the middle vessels appeared in only 93.3% +/- 4.6 of the cases. The superior group included on each side 4 arteries in males and 5 in females; the middle group presented only 1 artery on each side in both males and females, and the inferior group exhibited on each side 2 arteries in males and 1 artery in females. The most variable group was the middle one, the aortic origin being the most frequent but with a relatively low incidence (53.3% +/- 9.1 on the right and 46.7% +/- 9.1 on the left). The superior group originated from the posterior branch of the ipsilateral inferior phrenic artery in 83.3% +/- 6.8 on the right and 80% +/- 7.3 on the left. The arteries of the inferior group were branches of the ipsilateral renal artery in 70% +/- 8.4 on the right and 50% +/- 9.1 on the left. The origin of the middle suprarenal arteries from the trunk of the inferior phrenic artery on both sides (26.7% +/- 8.1 on the right and 36.7% +/- 8.8 on the left) should be considered relevant. The anatomical findings warrant a further investigation for the identification, illustration and nomenclature of arterial anatomicosurgical segments. PMID- 11035647 TI - [Scholars in the German Anatomical Institute: longitudinal study concerning the number, gender, personnel positions and the academic education]. PMID- 11035648 TI - Barbara, what's a nice girl like you doing writing an article like this? : the scientific basis of folk remedies for colds and flu. PMID- 11035649 TI - Thromboembolic disease : can echocardiography assist management? PMID- 11035650 TI - Airway obstruction in severe COPD. PMID- 11035651 TI - The labyrinth of asthma therapy: lost in the choices. PMID- 11035652 TI - Fertility in patients with cystic fibrosis. PMID- 11035653 TI - Sublingual capnometry : in search of its holy grail. PMID- 11035654 TI - Pathophysiology of impaired right and left ventricular function in chronic embolic pulmonary hypertension: changes after pulmonary thromboendarterectomy. AB - STUDY OBJECTIVES: This study sought to evaluate the pathophysiology of left and right heart failure in patients with chronic thromboembolic pulmonary hypertension (CTEPH) who were hospitalized to undergo pulmonary thromboendarterectomy (PTE). DESIGN: Thirty-nine patients (16 women and 23 men; mean +/- SD age, 55+/-12 years) with severe CTEPH were examined before and 13+/-8 days after PTE by way of transthoracic echocardiography and right heart catheterization. MEASUREMENTS AND RESULTS: Examination results confirmed in all cases that before surgery the right ventricles were enlarged and systolic function was impaired. Moderate to severe tricuspid valve regurgitation was observed. Left ventricular eccentricity indexes reflected a leftward displacement of the interventricular septum. End-diastolic left ventricular size and systolic function had decreased, and the left ventricular filling pattern showed impaired diastolic function. After surgery, mean pulmonary artery pressure was significantly lower (48+/- 10 mm Hg vs. 25+/-7 mm Hg; p<0.05). The calculated end diastolic and end-systolic right ventricular areas had decreased: 30+/-7 cm(2) vs 21 +/-5 cm(2) (p<0.05) and 24+/-6 cm(2) vs. 14+/-4 cm(2) (p<0.05), respectively. Right ventricular fractional area change had increased (20+/-7% vs. 33+/-8%; p<0.05). Most of the patients exhibited a marked decrease in the severity of tricuspid regurgitation. Septal motion, left ventricular systolic function, and diastolic filling pattern returned to normal values (early to late diastolic left ventricular inflow ratio, 0.70+/-0.33 vs. 1.35+/-0.51; p<0.05). The mean cardiac index also improved (2.7+/-0.6 L/min/m(2) vs. 3.7+/-0.8 L/min/m(2)). CONCLUSIONS: In CTEPH, functions are impaired in the right as well as the left ventricles of the heart. Improved lung perfusion and the reduction of right ventricular pressure overload are direct results of PTE, which in turn bring a profound reduction of right ventricular size and a recovery of systolic function. Normalization of interventricular septal motion as well as improved venous return to the left atrium lead to a normalization of left ventricular diastolic and systolic function, and the cardiac index improves. PMID- 11035655 TI - Admission serum potassium in patients with acute myocardial infarction: its correlates and value as a determinant of in-hospital outcome. AB - STUDY OBJECTIVES: Although controversial, hypokalemia (LK) in patients with acute myocardial infarction (MI) is thought to predict increased in-hospital morbidity, particularly cardiac arrhythmias, and mortality. Also, the mechanism of low serum potassium in the setting of MI has not been delineated. We evaluated the frequency, attributes, and outcome, and speculated on the mechanism of LK in patients with MI. DESIGN: This was a prospective cross-sectional study of 517 consecutive patients with MI admitted to the coronary care unit (CCU). Serum potassium was measured in the emergency department and repeatedly thereafter throughout hospitalization, and was used in the analysis, along with a large array of clinical and laboratory variables. RESULTS: The patients were allocated to a LK and a normokalemic (NK) cohort, based on the emergency department serum potassium measurement. The 41 patients with LK (3.16+/-0.24 mEq/L; 7.9% of total) were comparable on admission in their baseline assessment to the 476 patients with normal serum potassium (4.28+/-0.56 mEq/L), except for lower emergency department magnesium (1.48+/-0.15 mg/dL vs. 1.96+/-0.26 mg/dL; p = 0.0005) and earlier presentation after onset of symptoms (3.0+/-4.1 h vs. 4.4+/- 6.2 h; p = 0.05). There was a poor correlation between serum potassium and magnesium on admission (r = 0.14). Peak creatine kinase (CK) and myocardial isomer of CK were higher in the LK patients (3,870+/-3, 840 IU/L vs. 2,359+/-2,653 IU/L [p = 0.018] and 358+/-312 IU/L vs. 228 +/- 258 IU/L [p = 0.013], respectively). Management of the two cohorts was the same, except for a higher rate of use of magnesium (14.6% vs. 4.6%; p = 0.007), serum potassium supplements (90.2% vs 43. 1%; p = 0.000005), and antiarrhythmic drugs (78.0% vs 50.4%; p = 0. 0007) in the LK patients. No difference was detected between the LK and NK patients in total mortality (24.4% vs. 18.3%; p = 0.34), cardiac mortality (17.1% vs. 15.3%; p = 0.52), atrial fibrillation (14.6% vs 13.9%; p = 0.89), and ventricular tachycardia (22.0% vs. 16.0%; p = 0.32), but ventricular fibrillation (VF) occurred more often (24.4% vs 13.0%; p = 0.04) in the LK patients. However, proportions of VF occurring in the emergency department, CCU, or wards in the two cohorts were not different, but they were higher during the time interval prior to emergency department admission in LK patients (17.1% vs 2.1%; p = 0.00001). CONCLUSIONS: LK is seen in approximately 8% of patients with MI in the emergency department; LK is associated with low emergency department magnesium, and low serum potassium levels in the CCU and throughout hospitalization. LK has no relationship to preadmission use of diuretics, it is associated with early presentation to the emergency department, and it is not a predictor of increased morbidity or mortality. PMID- 11035656 TI - Incidence, predictive factors, and prognostic significance of supraventricular tachyarrhythmias in congestive heart failure. AB - BACKGROUND: The incidence, predictive factors, morbidity, and mortality associated with the development of supraventricular tachyarrhythmias (SVTs) in patients with congestive heart failure (CHF) are poorly defined. METHODS: In the Digitalis Investigation Group trial, patients with CHF who were in sinus rhythm were randomly assigned to digoxin (n = 3,889) or placebo (n = 3,899) and followed up for a mean of 37 months. Baseline factors that predicted the occurrence of SVT and the effects of SVT on total mortality, stroke, and hospitalization for worsening CHF were determined. RESULTS: Eight hundred sixty-six patients (11.1%) had SVT during the study period. Older age (odds ratio [OR], 1.029 for each year increase in age; p = 0.0001), male sex (OR, 1.270; p = 0.0075), increasing duration of CHF (OR, 1.003 for each month increase in duration of CHF; p = 0.0021), and a cardiothoracic ratio of > 0.50 (OR, 1.403; p = 0.0001) predicted an increased risk of experiencing SVT. Left ventricular ejection fraction, New York Heart Association functional class, and treatment with digoxin vs placebo were not related to the occurrence of SVT. After adjustment for other risk factors, development of SVT predicted a greater risk of subsequent total mortality (risk ratio [RR] = 2.451; p = 0.0001), stroke (RR = 2.352; p = 0.0001), and hospitalization for worsening CHF (RR = 3. 004; p = 0.0001). CONCLUSION: In CHF patients in sinus rhythm, older age, male sex, longer duration of CHF, and increased cardiothoracic ratio predict an increased risk for experiencing SVT. Development of SVT is a strong independent predictor of mortality, stroke, and hospitalization for CHF in this population. Prevention of SVT may prolong survival and reduce morbidity in CHF patients. PMID- 11035657 TI - Human herpesvirus 8 is not associated with sarcoidosis in Japanese patients. AB - STUDY OBJECTIVES: The etiology of sarcoidosis remains unknown, but recently it was reported that human herpesvirus 8 (HHV-8) may be detected in sarcoid tissue in a high proportion of patients. This study was performed to determine whether HHV-8 is implicated in sarcoidosis in Japan. MATERIALS AND METHODS: Peripheral blood mononuclear cells (PBMCs) were obtained from 100 patients with sarcoidosis and 100 healthy donors living in central Japan. Additionally, 19 samples of sarcoid tissue, 10 of tuberculous tissue, and 10 of lung cancers were examined. DNA was extracted from PBMC or tissue samples, and a hemi-nested polymerase chain reaction assay was performed for HHV-8 detection. RESULTS: In the PBMC study, the detection rates for HHV-8 in patients with sarcoidosis and in normal donors were 2% and 1%, respectively, the difference not being significant (p>0.99). In the tissue study, HHV-8 was detected in 10.5% of sarcoid and in 15% of nonsarcoid tissues, again not a significant difference (p>0.99). CONCLUSIONS: From these results, we conclude that HHV-8 is not implicated in sarcoidosis in Japanese patients. PMID- 11035658 TI - Diagnosing sarcoidosis using endosonography-guided fine-needle aspiration. AB - STUDY OBJECTIVES: The ability to diagnose sarcoidosis cytologically has been reported previously, but the method is rarely used. Endoscopic ultrasonography (EUS) is a sensitive technique for detecting mediastinal lymph nodes, which in addition provides an opportunity to carry out guided fine-needle aspiration (FNA) cytology. We report herein on the use of EUS-FNA in the diagnosis of sarcoidosis. PATIENTS AND METHODS: Nineteen patients with suspected sarcoidosis were investigated using EUS-FNA with a linear echoendoscope and a 22-gauge Hancke Vilman needle. MEASUREMENTS AND RESULTS: In all 19 patients, EUS revealed enlarged mediastinal lymph nodes (mean size, 2.4 cm), located subcarinally (n = 15), in the aortopulmonary window (n = 12), or in the lower posterior mediastinum (n = 5). The nodes had an isoechoic or hypoechoic appearance, with atypical vessels in five cases. The amount of aspirate obtained using EUS-FNA was adequate in all patients, and contained blood in excess of normal in some, indicating a high degree of vascularity. Cytology demonstrated epithelioid cell granuloma formation, suggesting sarcoidosis. Mycobacterial cultures were negative in all of the patients except one, in whom the final diagnosis was tuberculosis. The specificity and sensitivity of EUS-FNA in the diagnosis of sarcoidosis were 94% and 100%, respectively. CONCLUSIONS: EUS of mediastinal lymph nodes in sarcoidosis reveals certain characteristic features. However, it is not capable of differentiating the lesions from tuberculosis or malignancy. EUS-FNA is a safe and sensitive method of aspirating material for cytology and mycobacterial cultures. We believe it will provide a useful alternative in the diagnosis of sarcoidosis. PMID- 11035659 TI - Fine-needle aspiration cytologic technique for lung cancer has a high potential of malignant cell spread through the tract. AB - BACKGROUND: Fine-needle aspiration cytologic technique (FNAC), a method to detect malignancy for undetermined pulmonary nodules, may have a high potential to spread malignant cells from the tumor to the pleural cavity. OBJECTIVE: The authors assessed malignant cell spread through the needle tract following FNAC for peripheral lung carcinoma. MATERIALS AND METHODS: Lung lobes resected from 20 patients during the treatment of lung carcinoma were examined. The visceral pleura over the lung carcinoma was irrigated by heparinized saline solution to clean the surface, and then irrigated before FNAC and irrigated following FNAC to collect cells on the visceral pleura. FNAC was performed once for each tumor. Papanicolau's method was employed for cytologic examination. RESULTS: There were 15 specimens of adenocarcinoma, 4 specimens of squamous cell carcinoma, and 1 specimen of atypical carcinoid. The maximum diameter of the specimens ranged from 10 to 60 mm (median, 25 mm). Pleural indentation was observed in 15 samples. All results of FNAC were positive and matched the histologic diagnosis. Pre-FNAC specimens revealed a positive malignancy rate of 10% (2 of 20), but post-FNAC specimens had a rate of 60% (12 of 20; p = 0.002) CONCLUSION: FNAC has the potential to spread malignant cells to the pleural space. Further study is needed to determine the clinical significance of the spread of malignant cells in the pleural space. PMID- 11035660 TI - Prognosis and recurrent patterns in bronchioloalveolar carcinoma. AB - STUDY OBJECTIVE: Bronchioloalveolar carcinoma (BAC) is an uncommon pulmonary neoplasm with various radiologic and clinical presentations. In this article, we analyze the initial radiologic findings, TNM stagings, surgical types, and radiologic features of recurrence, and correlate them with patient survival. DESIGN: A retrospective review of 93 patients who underwent resection for BAC from February 1989 to May 1999. PATIENTS: There were a total of 153 patients with BAC diagnosed during this period. Among them, 60 patients (39.2%) had diffuse disease and received medical therapy only, and the remaining 93 patients (60.8%), who had localized disease, underwent surgical resection. Patients who received surgical resection were enrolled in this study. MEASUREMENTS: Data regarding demographics, presentation symptoms, initial radiologic features, surgical type, tumor staging, recurrence status, radiologic patterns of recurrence, and survival were obtained from all patients. RESULTS: Female patients were significantly younger than male patients. Patients who were female, nonsmoking, undergoing curative surgery, lobectomy, or bilobectomy, and with early tumor staging and no nodal involvement had a better prognosis. Patients with a right lung tumor had a longer survival than those with a left lung tumor, with borderline significance. Among those who suffered from recurrent diseases, a second resection yielded a better survival. Multivariate analysis showed curative surgery, initial surgical type, recurrence status, radiologic patterns of recurrence, and duration from surgical resection to recurrence all had a significant impact on survival. CONCLUSIONS: Those patients with localized, early-stage BAC who underwent curative surgery had a better survival. Patients with localized recurrence after the initial surgery warranted a second resection. Those with a diffuse radiologic pattern of recurrence and/or early recurrence had a worse prognosis. PMID- 11035661 TI - Serum levels of vascular endothelial growth factor dependent on the stage progression of lung cancer. AB - STUDY OBJECTIVE: In lung cancer, vascular endothelial growth factor (VEGF) is an important cytokine and is correlated with tumor vessel density, malignant pleural effusions, and coagulation-fibrinolysis factors in vitro. We investigated the correlation between serum VEGF level and stage progression in lung cancer to study the predicted value of VEGF level. We also studied whether coagulation fibrinolysis factors and PaO(2) levels, which are also important factors for the prediction of the clinical course, are correlated with VEGF. METHODS: Forty-nine patients with lung cancer were investigated prospectively. VEGF levels of sera and malignant effusions, and plasma concentrations of coagulation-fibrinolysis factors were measured by enzyme-linked immunosorbent assay. We measured PaO(2) levels in all patients at rest. RESULTS: Serum levels of VEGF were increased significantly according to stage progression. Additionally, plasma concentrations of D dimer, thrombin-antithrombin complex (TAT), and tissue plasminogen activator/plasminogen activator inhibitor type I complex were elevated significantly according to stage progression. The serum VEGF level had a significant positive correlation with the TAT and D dimer levels. Serum VEGF levels had a significant negative correlation with PaO(2) levels. The incidence of cerebral vascular disorder was significantly higher in the patients with systemic hypoxemia than in those without (p<0.05). Mean VEGF levels in malignant effusions in eight patients (five with pleural effusions, two with pericardial effusions, and one with both) were extremely high, especially in pericardial effusions ([mean +/- SD] pleural effusions, 531.9+/-285.4 pg/mL; pericardial effusion, 3,071.6+/-81.3 pg/mL). CONCLUSION: We predict that in lung cancer, VEGF production and the abnormality of the coagulation-fibrinolysis system differ depending on the stage of progression of disease. Serum VEGF levels would be affected by PaO(2) levels in lung cancer. PMID- 11035662 TI - Survival in synchronous vs. single lung cancer: upstaging better reflects prognosis. AB - OBJECTIVE: To define prognostic parameters for patients with synchronous non small cell lung cancer (NSCLC). DESIGN: Retrospective study of period from 1970 through 1997. PATIENTS: Patients with a single (n = 2,764) and synchronous NSCLC (n = 85) who underwent pulmonary resection. METHODS: All tumors were classified postsurgically, and the tumors of the patients with synchronous lung cancer were staged separately. The most advanced tumor was used for comparison. Actuarial survival time was estimated, and risk factors influencing survival were evaluated. Patients who died within 30 days of surgery were excluded. MEASUREMENT AND RESULTS: Five-year survival for single NSCLC was 41% and for synchronous lung cancer it was 19%. The relative risk of death for patients with synchronous lung cancer was 1.75, compared to that for patients with single lung cancer. The most advanced tumor in synchronous cancer was a significant predictor of survival (p<0.005). The survival of patients with synchronous lung cancer in which the most advanced tumors were stage I (n = 40) and stage II (n = 27) was not different from that of patients with stage II (n = 834) and stage IIIA (n = 405) single lung cancer, respectively. CONCLUSION: The poorer survival of patients with synchronous NSCLC is confirmed and quantified. The stage of the most advanced tumor was the best predictor of prognosis. The prognosis of patients with synchronous NSCLC resembles the prognosis of patients with a single lung cancer of a higher stage. Upstaging in synchronous lung cancer is recommended on the basis of these observations. PMID- 11035663 TI - Implantation of ultraflex nitinol stents in malignant tracheobronchial stenoses. AB - STUDY OBJECTIVE: To assess the uncovered Ultraflex nitinol stent (Boston Scientific; Natick, MA) for its efficacy and safety. DESIGN, SETTING, AND PATIENTS: Between October 1997 and October 1998, we carried out a prospective multicenter study at six hospitals in Japan. Fifty-four Ultraflex stents were inserted in 34 patients with inoperable malignant airway stenosis using a flexible and/or a rigid bronchoscope under fluoroscopic and endoscopic visualization. MEASUREMENTS AND RESULTS: Clinical, endoscopic examination, and pulmonary function on days 1, 30, and 60 after stent implantation showed improvement. In 19 patients (56%), stent implantation was performed as an emergency procedure because of life-threatening tracheobronchial obstruction. Immediate relief of dyspnea was achieved in 82% of the patients. The dyspnea index improved significantly after implantation (before vs. days 1, 30, and 60; p<0.001). Significant improvements were observed in obstruction of airway diameter (81+/-15% before vs. 14+/-17% on day 1, 12+/-12% on day 30, and 22+/-28% on day 60; p<0.001). Vital capacity (VC), FEV(1), and peak expiratory flow (PEF) increased significantly after stent implantation: before vs immediately after VC (p<0.01), FEV(1) (p<0.001), and PEF (p<0.05). The main complications were tumor ingrowth (24%) and tumor overgrowth (21%). After coagulation with an Nd-YAG laser or argon plasma coagulation, mechanical coring out using the bevel of a rigid bronchoscope was necessary in patients showing tumor ingrowth or overgrowth. Removal and reposition were possible in case of misplacement. There were no problems with migration and retained secretions. The median survival time of patients was 3 months. The 1-year survival rate was 25.4%. CONCLUSIONS: In this study of the Ultraflex nitinol stent, we have demonstrated that patients were relieved from dyspnea, which contributed to improved quality of life, with minimal complications. This stent can be used safely, even in the subglottic region. Owing to its excellent flexibility and biocompatibility, the stent is also indicated in certain complicated situations, eg, narrow stenosis, hourglass stenosis, curvilinear stenosis, bilateral mainstem bronchial stenoses, and long stenosis of varying diameters. PMID- 11035664 TI - Adenoviral-mediated p53 gene transfer to non-small cell lung cancer through endobronchial injection. AB - OBJECTIVE: The objective was to determine the degree of toxicity and antitumor activity following bronchoscopic injection of an adenoviral-mediated p53 gene (Adp53) into tumors causing airway obstruction. DOSING: This was a subset analysis of a phase I dose escalation trial. SETTING: Patients were treated in the outpatient clinics at the University of Texas (MD Anderson Cancer Center, Houston, TX) and at Medical City Dallas Hospital (US Oncology, Dallas, TX). PATIENTS: Twelve patients (median age, 60 years) with advanced endobronchial non small cell lung cancer (NSCLC) (squamous cell carcinoma, six patients; adenocarcinoma, six patients) were entered into trial. The median tumor area was 5 x 3.2 cm. All patient tumors contained a p53 gene mutation. INTERVENTIONS: Adp53 (dose range, 1 x 10(6) to 1 x 10(11) plaque-forming units) was administered by bronchoscopic intratumoral injection once every 28 days. MEASUREMENTS AND RESULTS: Toxicity attributed to the Adp53 vector was minimal. Six of the 12 patients had significant improvement in airway obstruction, and 3 patients met the criteria for partial response. CONCLUSIONS: Direct bronchoscopic injection of Adp53 into endobronchial NSCLC is safe, with acceptable levels of toxicity. The initial clinical results demonstrating relief of airway obstruction warrant further clinical investigation. PMID- 11035665 TI - A polymorphism in the tumor necrosis factor-alpha gene promoter region may predispose to a poor prognosis in COPD. AB - STUDY OBJECTIVES: To determine whether the adenine (A)-guanine (G) substitution polymorphism at position - 308 on the tumor necrosis factor-alpha gene confers susceptibility to COPD or to the development of a more severe form of disease. DESIGN: A cross-sectional study was undertaken to compare the frequency of the A allele in a group of 106 patients with COPD with that in a control population (n = 99). Patients were followed up prospectively for a period of 2 years. PARTICIPANTS AND SETTING: Participants included 106 COPD patients recruited from a respiratory outpatient clinic and 99 control subjects recruited from patients admitted for cardiac catheterization. MEASUREMENTS AND RESULTS: DNA was extracted from venous blood, and each subject was genotyped for the polymorphism by polymerase chain reaction amplification and restriction digestion using Nco1. There was no increased frequency of the A allele in patients compared to control subjects. AA homozygous patients had less reversible airflow obstruction (p<0.05) and a significantly greater mortality (both all-cause and respiratory deaths) on follow-up (p<0.001), despite a shorter cigarette smoking history. CONCLUSIONS: This study suggests that homozygosity for this A allele predisposes to more severe airflow obstruction and a worse prognosis in COPD. PMID- 11035666 TI - Maximal inspiratory flow rates in patients with COPD. AB - OBJECTIVES: To assess the relevance of maximal inspiratory flow rates (MIFR) in the assessment of airway obstruction in COPD. SETTING: University teaching hospital. PARTICIPANTS: Ten consecutive COPD patients (O group; mean [+/- SD] age, 58.5+/-8.3 years) and 10 matched healthy subjects (H group; mean age, 58.7+/ 7.4 years). MEASUREMENTS: Lung volumes, FEV(1), specific airway conductance, single-breath lung diffusing capacity, MIFR, and maximal expiratory flow rates (MEFR). RESULTS: Mean FEV(1)/vital capacity (VC) was 74.7% in the H group and 37.8% in the O group (p<0.001). Total lung capacity was higher (p<0.001) in the O group compared with the H group. Lung diffusing capacity was less than half in the O group compared with the H group (p<0.001). MEFR at all lung volumes were lower in the O group (p<0.001). MIFR were comparable in the two groups, except at 25% inspired VC, where MIFR were lower in the O group (p< 0.05). CONCLUSION: MIFR are less sensitive than MEFR to detect airway obstruction in COPD patients. Yet, the interest of MIFR lay in the possibility to separate intrinsic from extrinsic involvement of airways. A normal MIFR associated with low MEFR, as in the present study, suggests either a lack of parenchymal support, an increased collapsibility of the airways, or a reversible peripheral airway narrowing. A fixed, generalized airway narrowing would be associated with a decrease of both MIFR and MEFR. PMID- 11035667 TI - Geographic variations in prevalence and underdiagnosis of COPD: results of the IBERPOC multicentre epidemiological study. AB - OBJECTIVES: To ascertain the prevalence, diagnostic level, and treatment of COPD in Spain through a multicenter study comprising seven different geographic areas. DESIGN AND PARTICIPANTS: This is an epidemiologic, multicenter, population-based study conducted in seven areas of Spain. A total of 4,035 men and women (age range, 40 to 69 years) who were randomly selected from a target population of 236,412 subjects participated in the study. INTERVENTIONS: Eligible subjects answered the European Commission for Steel and Coal questionnaire. Spirometry was performed, followed by a bronchodilator test when bronchial obstruction was present. RESULTS: The prevalence of COPD was 9.1% (95% confidence interval [CI], 8.1 to 10.2%), 15% in smokers (95% CI, 12.8 to 17.1%), 12.8% in ex-smokers (95% CI, 10.7 to 14.8%), and 4.1% in nonsmokers (95% CI, 3.3 to 5.1%). The prevalence in men was 14.3% (95% CI, 12.8 to 15. 9%) and 3.9% in women (95% CI, 3.1 to 4.8%). Marked differences were observed between sexes in smoking; the percentage of nonsmokers was 23% in men and 76.3% in women (p<0.0001). The prevalence of COPD varied among the areas, ranging from 4.9% (95% CI, 3.2 to 7.0%) in the area of the lowest prevalence to 18% (95% CI, 14.8 to 21.2%) in the area of the highest. There was no previous diagnosis of COPD in 78.2% of cases (284 of 363). Only 49.3% of patients with severe COPD, 11.8% of patients with moderate COPD, and 10% of patients with mild COPD were receiving some kind of treatment for COPD. Multivariate analysis showed that individuals had a higher probability of having received a previous diagnosis of COPD if they lived in urban areas, were of male gender, were > 60 years old, had higher educational levels, had > 15 pack year smoking history, or had symptoms of chronic bronchitis. CONCLUSIONS: COPD is a very frequent disease in Spain, and presents significant geographic variations and a very low level of previous diagnosis and treatment, even in the most advanced cases. PMID- 11035668 TI - Controlled short-term trial of fluticasone propionate in ventilator-dependent patients with COPD. AB - BACKGROUND: There is no agreement about the efficacy of systemic corticosteroids in patients with COPD, but corticosteroids often are employed during exacerbations of the disease. The use of systemic or inhaled corticosteroids in patients in stable condition is even more controversial, even though the more severely affected patients seem to respond better. Unfortunately, in this subset of patients, the use of forced expiratory maneuvers frequently fails to detect significant functional response. STUDY OBJECTIVES: We evaluated the short-term effects of an inhaled corticosteroid, fluticasone propionate (FP), on FEV(1) and on the mechanical properties of patients in stable condition with severe COPD and chronic hypercapnic respiratory failure who were receiving long-term ventilatory support. This allowed us to measure respiratory mechanics (RM) passively, thereby avoiding any problems linked with voluntary maneuvers. DESIGN: Randomized, placebo-controlled, crossover study. SETTING: A respiratory ICU. PATIENTS: Twelve hypercapnic COPD patients (mean [+/- SD] PaCO(2), 60+/-11 mm Hg; mean FEV(1), 13+/-5% predicted; and mean FEV(1)/FVC, 31 +/- 7%) were enrolled. INTERVENTIONS: A daily dose of 2,000 microg FP or placebo was administered via metered-dose inhaler during mechanical ventilation for 5 consecutive days. A washout of 72 h was allowed between regimens. MEASUREMENTS: End-expiratory and end-inspiratory airway occlusions were performed to assess static intrinsic positive end expiratory pressure (PEEPi,st), static compliance of the respiratory system (Cst,rs), maximal respiratory resistance (Rmax, rs), and minimal respiratory resistance (Rmin,rs). The bronchodilator response also was assessed by FEV(1) level. RESULTS: No significant changes were found in RM after administration of the placebo. By day 6, FP had induced the following significant decreases: PEEPi,st, 4.3+/-2.4 to 3.1+/-1.7 cm H(2)O (p<0.01); Rmax,rs, 19.0+/-6.5 to 14.6+/ 6 cm H(2)O/L/s (p<0.001); and Rmin,rs, 14.8+/-4.2 to 10.5+/-3.4 cm H(2)O/L/s (p<0.001). The Cst,rs and the effective additional resistance of the respiratory system did not change significantly, the latter suggesting that the major effect of FP was on the airway caliber (Rmin,rs). FEV(1) changes significantly (p<0.01) underestimated the bronchodilator response, as compared with changes in Rmin,rs. CONCLUSIONS: We conclude that in patients in stable condition with severe COPD and chronic hypercapnic respiratory failure, a brief trial of FP may induce a bronchodilator response, mainly related to a reduction in airway resistances, that is not detected by the usual pulmonary function tests. PMID- 11035669 TI - Latex allergy in children on home mechanical ventilation. AB - STUDY OBJECTIVE: Determining the incidence of latex allergy in children receiving home mechanical ventilation. BACKGROUND: The prevalence of latex allergy in the general population ranges from 0.1 to 1.0%. However, in patients with spina bifida and other chronic medical conditions associated with repeated exposure to latex, the prevalence may be as high as 60%. Children receiving home mechanical ventilation are frequently exposed to latex products. Therefore, we hypothesized that these children would be at increased risk for latex allergy. DESIGN: Fifty seven children receiving home mechanical ventilation (31 boys, 26 girls; mean [+/ SD] age, 7.8+/-6.6 years; range, 0.3 to 23.2 years) were enrolled. A radioallergosorbent test (RAST) for latex was administered and serum IgE levels were obtained in all patients. RESULTS: Seventeen patients (29.8%) were found to have a positive RAST for latex. Patients with latex allergy had required mechanical ventilation for an average of 6.1+/-4.1 years vs. 5.5+/-5.4 years (p = 0.69; not significant) in those without latex allergy. Eleven of 17 patients (64.7%) had elevated serum IgE levels in the group with latex allergy vs only 14 of 40 patients (35.0%) in the group with a negative latex RAST (p = 0.04; odds ratio, 3.4). CONCLUSION: We conclude that there is a high incidence of latex allergy in children requiring home mechanical ventilation. We speculate that screening all children receiving home mechanical ventilation may lead to the identification of patients with previously undiagnosed latex allergy and the prevention of untoward reactions from exposure to latex. PMID- 11035670 TI - Hemodynamic effects of noninvasive bilevel positive airway pressure on patients with chronic congestive heart failure with systolic dysfunction. AB - BACKGROUND AND STUDY OBJECTIVES: Noninvasive positive airway pressure may play a significant role in treating patients with congestive heart failure (CHF). We tested our hypothesis that noninvasive bilevel positive airway pressure improves left ventricular performance in patients with chronic CHF secondary to severe systolic dysfunction. OBJECTIVES: To determine the cardiac performance of patients using bilevel positive airway pressure, and to describe the hemodynamic effects of bilevel positive airway pressure and its use as a therapeutic adjunct in these patients. DESIGN: Prospective, cohort, nonrandomized study. SETTING: Outpatient medicine clinic. PATIENTS: Fourteen patients (9 men and 5 women) with stable chronic CHF with left ventricular ejection fraction < or =35%; mean age was 60.6 years (range, 43 to 87 years). INTERVENTIONS: Bilevel positive airway pressure via nasal mask at an inspiratory pressure of 5 cm H(2)O and an expiratory pressure of 3 cm H(2)O on spontaneous mode at room air for 1 h. MEASUREMENTS AND RESULTS: Myocardial performance and changes were measured using clinical and echocardiographic parameters. Baseline clinical and echocardiographic parameters were compared with the same parameters after 1 h of bilevel positive airway pressure. Statistically significant (p<0.05, Wilcoxon matched pair signed-rank test) decreases were noted in these mean values: systolic BP from 136.21 to 124.14 mm Hg (p = 0.008), heart rate from 85.07 to 74.71 beats/min (p = 0.002), respiratory rate from 23.07 to 15.43 breaths/min (p = 0.001), and systemic vascular resistance from 1671. 46 to 1236.27 dyne. s. cm(3) (p = 0.001). Statistically significant increases were noted in these mean values: cardiac output from 5.09 to 6.37 L/min (p = 0.004), ejection fraction from 28.71% to 34.36% (p = 0.001), and end-diastolic volume from 224.36 to 246.21 mL (p = 0.045). CONCLUSION: Bilevel positive airway pressure has excellent potential for improving left ventricular performance of patients with chronic CHF secondary to severe systolic dysfunction. PMID- 11035671 TI - Constant vs. automatic continuous positive airway pressure therapy: home evaluation. AB - STUDY OBJECTIVES: To compare the efficacy and patient tolerance, compliance, and preference between auto-continuous positive airway pressure (CPAP) and constant CPAP. DESIGN: Single-blinded, crossover, cohort study of consecutive patients with obstructive sleep apnea syndrome, with two treatment periods of 2 months each. PATIENTS: Twenty-five patients (22 men, 3 women) with sleep apnea syndrome confirmed by ambulatory polysomnography. MEASUREMENTS AND INTERVENTIONS: After baseline polysomnography, patients underwent in-laboratory polysomnography for titration of constant CPAP. The order of treatment was randomly allocated, either auto-CPAP and then constant CPAP, or vice versa. The auto-CPAP pressure range was 6 to 16 cm H(2)O. At the end of each 2-month period, patients underwent a control ambulatory polysomnography and received a questionnaire on subjective well-being and device evaluation. Duration of use was checked through CPAP device monitoring. RESULTS: No differences were found in apnea/hypopnea index (constant CPAP, 9.7+/-1.9 events/h vs auto-CPAP, 10.6+/-9.3 events/h), awakening/arousal index (constant CPAP, 13.7 +/- 8.0 events/h vs auto-CPAP, 15.5 +/- 8.9 events/h), slow-wave sleep duration, nocturnal saturation, or complaint of daytime sleepiness. The mean pressure required was significantly lower during auto-CPAP than during constant CPAP (8.8+/-1.8 cm H(2)O vs. 9.7+/-2.6 cm H(2)O, respectively). Patient tolerance, compliance, and duration of use were similar with both treatments. CONCLUSIONS: Auto-CPAP is as effective as constant CPAP. A wide pressure range for auto-CPAP can be used in all patients, suggesting that, in the future, use of a broad pressure range in the auto-CPAP mode could obviate the need for the titration night. PMID- 11035672 TI - Association of body position with severity of apneic events in patients with severe nonpositional obstructive sleep apnea. AB - STUDY OBJECTIVE: To compare the severity of sleep apneic events occurring in the supine posture vs the severity of sleep apneic events occurring in the lateral posture in patients with severe obstructive sleep apnea (OSA). DESIGN: A retrospective analysis of apneic event variables in a group of 30 OSA patients who underwent a complete polysomnographic evaluation in our sleep disorders unit. PATIENTS: Thirty patients with severe OSA (respiratory disturbance index [RDI] = 70.1+/-18.2) who were nonpositional patients (NPP), ie, in whom the ratio of the supine RDI to the lateral RDI is < 2 (supine RDI = 85.7+/-11.7, lateral RDI = 64.8+/-17.3), and who had > or =30 apneic events in the lateral position and 30 apneic events in the supine position during sleep stage 2 were included in the study. MEASUREMENTS: For the 30 apneic events in each body position, the following variables were evaluated: apnea duration (ApDur), minimum desaturation (MinDes), Delta desaturation (Delta-Des), duration of arousal (DurArous), maximum snoring loudness (MaxSL), and Delta heart rate (Delta-HR). In addition, three other variables assessed as a ratio of ApDur (Rate-D = Delta-Des/ApDur, R-HR =Delta-HR/ApDur, and R-Arous = DurArous/ApDur) were also calculated. RESULTS: For all variables evaluated, apneic events occurring in the supine posture were significantly more severe than those apneic events occurring in the lateral posture during sleep stage 2. ApDur of both body postures correlated significantly with DurArous, Delta-HR, and MaxSL, but not with Delta-Des and MinDes. ApDur correlated linearly with DurArous for both postures. The slopes of the two regression lines were similar (p = 0.578) but the regression line intercept for the supine apneas was significantly higher than that of lateral apneas (p<0.0001). In addition, the average number of supine apneic events that did not end with an arousal was smaller than the average number of lateral apneic events not ending with an arousal (4.4+/-6.0 vs. 10.5+/-6.7, respectively; p< 0.0001). Also, only 4 of 900 (0.44%) apneic events analyzed in the lateral posture ended with an awakening (> 15 s), whereas in the supine posture, there were 37 (4.1%) such events (p<0.001). CONCLUSIONS: These results show that even in patients with severe OSA who have a high number of apneic events in the supine and lateral posture, the apneic events occurring in the supine position are more severe than those occurring while sleeping in the lateral position. Thus, it is not only the number of apneic events that worsen in the supine sleep position but, probably no less important, the nature of the apneic events themselves. PMID- 11035673 TI - Simple predictors of uvulopalatopharyngoplasty outcome in the treatment of obstructive sleep apnea. AB - STUDY OBJECTIVE: Our objective was to determine whether baseline polysomnography, cephalometry, and anthropometry data could predict uvulopalatopharyngoplasty (UPPP) success or failure. DESIGN: We retrospectively reviewed polysomnography, cephalometry, and anthropometry data from patients who underwent UPPP for obstructive sleep apnea (OSA). SETTING: A university medical center. PATIENTS: OSA was diagnosed by polysomnography in 46 patients who underwent UPPP surgery for their sleep disorder. INTERVENTIONS: UPPP surgery with/or without tonsillectomy. MEASUREMENTS AND RESULTS: The mean patient age was 43 years, and the mean body mass index was 32.5 kg/m(2). The mean presurgical apnea-hypopnea index (AHI) was 45, and the mean baseline nadir oxygen saturation was 81%. Successful surgery was defined as a reduction in AHI to < 10 or to < 20 with a 50% reduction from the patient's baseline AHI. Of the 46 patients, 16 were successfully treated and 30 did not respond to surgical treatment. A mandibular hyoid distance (MP-H) > 20 mm was found to be significantly (p = 0.05) predictive of failure of UPPP. When stepwise regression analysis was performed utilizing postsurgical AHI as the dependent variable and presurgical AHI, age, body mass index, baseline nadir O(2) saturation, and five cephalometric measurements as independent variables, MP-H distance significantly (r = 0.524; p = 0.01) correlated positively with postsurgical AHI. The distance between the superior point of a line-constructed plane of the sphenoidale (parallel to Frankfort horizontal) and a point at the intersection of the palatal plane perpendicular to the hyoid correlated negatively with postsurgical AHI (r = 0.586; p = 0.05). By creating a logistic model of this data, an MP-H distance < 21 mm, an angle created by point A to the nasion to point B < 3 degrees, and the presence of a baseline AHI < 38 enhanced the predictability of UPPP success. CONCLUSIONS: The presence of a baseline AHI < 38 and an MP-H < or = 20 mm, and the absence of retrognathia are predictors of improvement after UPPP. Based on these findings, we would advocate the continued evaluation of cephalometric measurements and careful consideration of surgical treatment options for OSA. PMID- 11035674 TI - Abbreviated method for assessing upper airway function in obstructive sleep apnea. AB - STUDY OBJECTIVES: Previous studies have shown that the level of flow through the upper airway in patients with obstructive sleep apnea (OSA) is determined by the critical closing pressure (Pcrit) and the upstream resistance (RN). We developed a standardized protocol for delineating quasisteady-state pressure-flow relationships for the upper airway from which these variables could be derived. In addition, we investigated the effect of body position and sleep stage on these variables by determining Pcrit and RN, and their confidence intervals (CIs), for each condition. DESIGN: Pressure-flow relationships were constructed in the supine and lateral recumbent positions (nonrapid eye movement [NREM] sleep, n = 10) and in the supine position (rapid eye movement [REM] sleep, n = 5). SETTING: University Hospital Antwerp, Belgium. PATIENTS: Ten obese patients (body mass index, 32.0+/-5.6 kg/m(2)) with severe OSA (respiratory disturbance index, 63.0+/ 14.6 events/h) were studied. INTERVENTIONS: Pressure-flow relationships were constructed from breaths obtained during a series of step decreases in nasal pressure (34.1+/-6.5 runs over 3.6+/-1.2 h) in NREM sleep and during 7.8+/-2.2 runs over 0.8+/-0.6 h in REM sleep. RESULTS: Maximal inspiratory airflow reached a steady state in the third through fifth breaths following a decrease in nasal pressure. Analysis of pressure-flow relationships derived from these breaths showed that Pcrit fell from 1.8 (95% CI, -0.1 to 2.7) cm H(2)O in the supine position to -1.1 cm H(2)O (95% CI, -1.8 to 0.4 cm H(2)O; p = 0.009) in the lateral recumbent position, whereas RN did not change significantly. In contrast, no significant effect of sleep stage was found on either Pcrit or RN. CONCLUSIONS: Our methods for delineating upper airway pressure-flow relationships during sleep allow for multiple determinations of Pcrit within a single night from which small yet significant differences can be discerned between study conditions. PMID- 11035675 TI - Effects of high-dose inhaled fluticasone propionate via spacer on cell-mediated immunity in healthy volunteers. AB - BACKGROUND: Systemic corticosteroids are known to alter cell-mediated immunity (CMI). However, the effects of inhaled steroids on CMI are unclear. We therefore sought to assess CMI following high-dose inhaled steroids in healthy subjects. METHODS: Ten healthy nonasthmatic subjects self-administered fluticasone propionate (FP), 440 microg bid, with a spacer device. CMI was assessed by delayed hypersensitivity skin testing to multiple antigens and in vitro by phytohemagglutinin (PHA) stimulation of peripheral blood T lymphocytes. Percentages of CD3(+), CD4(+), and CD3(+)CD8(+) cells expressing CD69(+) were determined by three-color flow cytometry. Studies were conducted before and after 4 weeks of FP treatment. RESULTS: After 4 weeks of FP treatment, two of nine subjects became anergic, whereas six of nine subjects had reduced skin responses (one subject was excluded). Mean total skin test score fell from 18.4+/-10.9 to 9.1 +/-7.2 mm (p = 0.02). There was a decline in tuberculin responses in all four subjects who were positive prior to FP treatment. Following FP treatment, the percentage of unstimulated (from control subjects receiving saline solution) CD3(+)CD4(+)CD69(+) cells declined from 14.8+/-4.2% to 8. 5+/-4.6% (p = 0.02) and the CD3(+)CD8(+)CD69(+) cells decreased from 29.7+/-12.7% to 17.1 +/-5.0% (p = 0.007). PHA stimulation produced significant increases in the percentage of CD3(+)CD4(+)CD69(+) cells before and after FP treatment (67.0+/-9.1%, p<0.02 before FP; 55.4+/-17.0%, p<0.02 after FP), and in the percentage of CD3(+)CD8(+)CD69(+) cells before and after treatment (79.7+/-9.3%, p<0.03 before FP; 71.2+/-11.4%, p = 0. 008 after FP). CONCLUSIONS: High doses of FP suppress the proportion of activated circulating T cells but do not affect the ability of T cells to respond to direct stimulation with PHA. However, depression of skin test responses to antigens following treatment with FP suggests an impairment of in vivo clinical manifestations of T-cell activation by a mechanism that requires further investigation. PMID- 11035676 TI - Effects of once-daily formoterol and budesonide given alone or in combination on surrogate inflammatory markers in asthmatic adults. AB - OBJECTIVES: We wished to evaluate the effects of once-daily combination therapy on surrogate inflammatory markers. METHODS: Fifteen patients with atopic persistent asthma were evaluated (mean age, 32.4 years; FEV(1), 75.2% predicted) in a randomized, double-blind, double-dummy, placebo-controlled crossover study with a 1-week placebo washout period, comparing the following once-daily nighttime treatments: (1) formoterol (FM), 12 microg, for 2 weeks and FM, 24 microg, for 2 weeks; or (2) budesonide (BUD), 400 microg, for 2 weeks and BUD, 800 microg, for 2 weeks; or (3) FM, 12 microg, plus BUD, 400 microg, for 2 weeks and FM, 24 microg, plus BUD, 800 microg, for 2 weeks. Adenosine monophosphate (AMP) bronchial challenge, exhaled nitric oxide (NO), and serum eosinophilic cationic protein (ECP) were evaluated at 12 h postdosing after administration of each placebo and after 2 and 4 weeks of each treatment. RESULTS: The results of AMP challenge (provocative concentration causing a 20% fall in FEV(1)) at 4 weeks showed significant (p<0.05) improvements after patients had received all active treatments compared to placebo (20 mg/mL), with FM plus BUD, 261 mg/mL, being superior (p<0.05) to FM alone, 82 mg/mL, but not to BUD, 201 mg/mL. NO and ECP showed significant (p<0.05) reductions compared to placebo with FM plus BUD or BUD alone but not with FM alone. Combination therapy was associated with optimal patient preference (rank order, FM plus BUD > FM > BUD; p<0.0005), highest domiciliary peak expiratory flow, and lowest rescue inhaler usage. All three treatments produced equivalent improvements in spirometry. CONCLUSIONS: Patients preferred once-daily combination therapy, but this had no greater effect on inflammatory markers than therapy with BUD alone. FM alone had no anti inflammatory activity but exhibited bronchoprotection. This emphasizes the importance of first optimizing anti-inflammatory control with inhaled corticosteroids before considering adding a regular long-acting beta(2)-agonist. PMID- 11035677 TI - Fertility in men with cystic fibrosis: an update on current surgical practices and outcomes. AB - BACKGROUND: Men with cystic fibrosis (CF) have bilateral absence of the vas deferens causing an obstructive azoospermia that is not amenable to surgical correction. Advances in the field of reproductive medicine allow for the procurement of viable sperm and facilitate fertilization and pregnancy in couples where the man has CF. OBJECTIVES: To describe patient anatomy and semen characteristics and to determine the pregnancy rates of couples in whom the male partner has CF and who have undergone microsurgical epididymal sperm aspiration coupled with in vitro technology, specifically intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective analysis. SETTING: Clinical department of urology and two reproductive medicine units. PATIENTS: Thirteen married men with CF who were referred for infertility. INTERVENTIONS: History, physical examination, semen analysis, transrectal and renal ultrasonography, CF mutation analysis, and microsurgical sperm aspiration coupled with ICSI. RESULTS: All 13 men had low volume azoospermia, absent vasa, and aplasia/hypoplasia of the seminal vesicles. CF mutation analysis was carried out in 11 of 13 men, and 9 of 11 were DeltaF508 homozygous. Eight men underwent microsurgical sperm aspiration, and their partners underwent one or more cycles of ICSI. Five couples (62.5%) achieved a pregnancy, with four couples delivering (three sets of twins and one singleton). CONCLUSIONS: CF in men is accompanied by bilateral vasal aplasia. The resultant obstructive azoospermia can be treated quite successfully with a combination of sperm aspiration and ICSI. It is important for physicians involved in the care of men with CF to convey the message that prospects for fatherhood are excellent with current technology. PMID- 11035678 TI - Right ventricular dysfunction in adult severe cystic fibrosis. AB - STUDY OBJECTIVES: This study sought to assess the extent of impairment of cardiac function in adult patients with end-stage cystic fibrosis (CF) and to examine the relationship between cardiovascular abnormalities and the degree of hypoxemia and hypercapnia. DESIGN AND SETTING: A retrospective study in a tertiary cardiac and CF center. PARTICIPANTS AND INTERVENTIONS: A total of 103 adult patients with end stage CF awaiting lung or heart and lung transplantation (mean age [+/- SD], 26+/ 7 years; 54 men) underwent Doppler echocardiography and arterial blood gas analysis (mean PaO(2), 54+/-10 mm Hg; mean PaCO(2), 47+/-8 mm Hg). The findings were compared to those of 17 healthy control subjects (mean age, 24+/-7 years; 13 men) who had no history of cardiac or pulmonary disease. MEASUREMENTS AND RESULTS: All patients were in sinus rhythm with a mean tachycardia of 112+/-18 beats/min (control subjects, 76+/-16; p<0.0001) and had a cardiac output of 5.3 L/min (control subjects, 4.3 L/min; p<0.04). In the patient group, the left ventricular (LV) dimensions, systolic and diastolic function, and wall thickness were all within normal limits. The mean amplitude of long-axis excursion in patients was normal at the LV site, but that of the right ventricular (RV) free wall was significantly reduced as compared with control subjects (1.6+/-0.4 vs. 2.2+/-0.4 cm, respectively; p<0.001), which was found to correlate with the degree of hypoxemia (r = 0.63; p<0.02) and hypercapnia (r = -0.68; p<0.01). RV diastolic function, which was represented by the relative isovolumic relaxation time to cardiac cycle length, was longer in patients than in control subjects (8.7+/-4.8% vs. 5.0+/-3.0%, respectively; p<0.03). The pulmonary flow acceleration time (90+/-22 vs 121+/-34 ms, respectively; p<0.01) and the systolic stroke distance (7.0+/-2.2 vs. 10.5+/-1.9 cm/s(2); p<0.001) were both lower than normal. CONCLUSIONS: This study confirms the presence of significant RV systolic and diastolic dysfunction in the setting of consistent tachycardia and increased cardiac output in adult CF patients with severe disease. No specific LV abnormalities were detected in these patients. PMID- 11035679 TI - Targeting aerosol deposition in patients with cystic fibrosis: effects of alterations in particle size and inspiratory flow rate. AB - STUDY OBJECTIVE: To determine if aerosolized medications can be targeted to deposit in the smaller, peripheral airways or the larger, central airways of adult cystic fibrosis (CF) patients by varying particle size and inspiratory flow rate. DESIGN: Randomized clinical trial. SETTING: Outpatient research laboratory. PATIENTS: Nine adult patients with CF. INTERVENTIONS: Patients inhaled an aerosol comprised of 3.68+/-0.04 microm saline solution droplets (two visits) or 1.01+/- 0.2 microm saline solution droplets (two visits) for 30 s, starting from functional residual capacity and breathing at a slow or faster inspiratory flow rate. On all visits, the saline solution was admixed with the radioisotope (99m)Tc. Immediately after inhalation, a gamma camera recorded the deposition pattern of the radioaerosol in the lungs. Deposition images were analyzed in terms of the inner:outer zone (I:O) ratio, a measure of deposition in an inner zone (large, central airways) vs. an outer zone (small airways and alveoli). MEASUREMENTS AND RESULTS: For the 3.68-microm aerosol, I:O ratios averaged 2.29+/ 1.45 and 2.54+/-1.48 (p>0.05), indicating that aerosol distribution within the lungs was unchanged while breathing at 12+/-2 L/min vs. 31+/-5 L/min, respectively. For the 1.01-microm aerosol, I:O ratios averaged 2.09+/-0.96 and 3.19+/-1.95 (p<0.05), indicating that deposition was predominantly in the smaller airways while breathing at 18+/-5 L/min and in the larger airways while breathing at 38+/-8 L/min, respectively. CONCLUSIONS: These results suggest that the targeted delivery of an aerosol to the smaller, peripheral airways or the larger, central airways of adult CF patients may be achieved by generating an aerosol comprised of approximately 1.0-microm particles and inspiring from functional residual capacity at approximately 18 L/min and approximately 38 L/min, respectively. PMID- 11035680 TI - Hemodynamic effects of epoprostenol in patients with systemic sclerosis and pulmonary hypertension. AB - STUDY OBJECTIVES: To determine the cause of pulmonary hypertension (PH) in systemic sclerosis (SSc) patients since PH can occur because of pulmonary arteriopathy, pulmonary parenchymal destruction, and left ventricular cardiac dysfunction. DESIGN AND SETTING: Consecutive case series in a university hospital. PATIENTS: Nine SSc patients with PH (mean pulmonary artery pressure, 41 mm Hg), with (n = 6) or without (n = 3) concomitant interstitial lung disease (ILD). METHODS: Acute infusion of epoprostenol was begun at 2 ng/kg/min and was titrated upward at a rate of 2 ng/kg/min every 30 min until symptomatic complications developed or pulmonary artery vascular resistance (PVR) was reduced by 50%. RESULTS: Eight of nine patients demonstrated a reduction of > or = 20% in PVR, suggesting that vasoreactivity is common despite the presence of significant ILD. A single patient had no response to infusion with unchanged hemodynamics and oxygenation. One patient developed hypoxemia as cardiac output increased, suggesting a worsening of ventilation/perfusion matching or the presence of an anatomic shunt. Acute pulmonary edema developed in one patient at an infusion rate of 6 ng/kg/min. The results of cardiac catheterization suggested that pulmonary edema was caused by SSc heart disease. CONCLUSION: SSc patients with ILD have diverse and sometimes multiple causes of PH that can be determined by short-term epoprostenol infusion. Beneficial effects can be obtained from epoprostenol despite extensive ILD. PMID- 11035681 TI - Alveolar hemorrhage in systemic lupus erythematosus: presentation and management. AB - AIM: To describe our experience with alveolar hemorrhage (AH) in systemic lupus erythematosus (SLE). METHODS: Review of medical records and pertinent medical literature using MEDLINE and reference lists from retrieved publications. PATIENTS: Seven patients with SLE admitted with episodes of AH (n = 11). RESULTS: Six patients were female, and one was male. Mean age at the time of AH was 31.1 years. Mean duration of SLE was 4.5 years. AH occurred within 3 weeks of SLE onset in two patients. Recurrent AH was observed in four patients. Six patients were already receiving treatment for SLE at the time of AH. All patients presented with dyspnea and new pulmonary infiltrates. Hemoptysis occurred in only 54%. All patients had BAL within 48 h of presentation. Temperature > or =39 degrees C (102.2 degrees F) accompanied 82% of episodes. Glomerulonephritis was the most common nonpulmonary SLE manifestation (74%). Treatment with empiric IV antibiotics was initiated in 10 episodes. Initial treatment included high-dose corticosteroids (prednisone, 1 to 3 mg/kg/d [n = 2]; or IV methylprednisolone, 1 g/d [n = 9], with or without oral cyclophosphamide, 2 to 3 mg/kg/d [n = 7]). Plasmapheresis (three to four sessions) was added in five episodes for persistent AH. All patients survived. CONCLUSIONS: AH may mimic pneumonia. Hemoptysis may not be evident. Infection must be aggressively excluded, especially since many patients with AH are already receiving immunosuppressive therapy. AH frequently recurs despite ongoing immunosuppression. Although high mortality rates have been reported with AH in SLE, we observed 100% survival. PMID- 11035682 TI - Is there an association between angiotensin-converting enzyme gene variants and chronic nonproductive cough? AB - BACKGROUND: It is unclear why some patients develop a chronic nonproductive cough. Angiotensin-converting enzyme (ACE) inactivates tussive peptides in the airways such as bradykinin and tachykinins. An insertion/deletion polymorphism in the ACE gene accounts for variation in ACE levels, and patients with the II genotype have lowest serum ACE levels compared with ID and DD genotypes. We hypothesized that the II genotype would be associated with increased risk of developing a chronic cough. MATERIALS AND METHODS: We recruited 47 patients (33 women), referred for evaluation of cough (median cough duration, 24 months; range, 2 to 240 months). Cough patients were evaluated using a comprehensive diagnostic protocol, and cough reflex sensitivity was measured using a capsaicin inhalation challenge. ACE genotyping was performed on DNA samples from patients using the polymerase chain reaction followed by agarose gel electrophoresis. ACE genotypes in patients with chronic cough were compared with those in 199 healthy control subjects. Serum ACE levels were determined using a colorimetric assay. RESULTS: Genotype frequencies for the ACE gene were similar between patients and control subjects. There was no correlation between capsaicin sensitivity and ACE genotypes or serum ACE levels. CONCLUSION: Susceptibility to develop chronic cough is not associated with ACE genotype. PMID- 11035683 TI - Bedside detection of retained tracheobronchial secretions in patients receiving mechanical ventilation: is it time for tracheal suctioning? AB - OBJECTIVE: To identify parameters that indicate retained secretions and the need for tracheal suctioning (TS) in patients receiving mechanical ventilation (MV). DESIGN: Prospective observational study. SETTING: A 14-bed medical ICU in a 946 bed university hospital. PATIENTS: Sixty-six consecutive patients receiving MV. INTERVENTIONS: Two successive tracheal suctions, TS1 and TS2, performed at a 2-h interval as usual patient care. Retained secretions were considered significant if the volume of secretions removed by TS2 was > 0.5 mL. MEASUREMENTS AND RESULTS: Variations between TS1 and TS2 of pulse oximetric saturation (SpO(2)), peak inspiratory pressure (Ppeak), tidal volume (VT), and Ramsay score were compared between patients with TS2 < or = 0.5 mL (group 1; n = 27) and patients with TS2 > 0.5 mL (group 2; n = 39). The presence of a sawtooth pattern on flow volume loop displayed on the monitor screen of the ventilator and of respiratory sounds heard over the trachea before TS2 were compared between the two groups. Variations of Ppeak, VT, SpO(2), and Ramsay score between TS1 and TS2 did not differ between the two groups. However, group 2 had a sawtooth pattern (82% vs 29.6%; p = 0.0001) and respiratory sounds (66.6% vs. 25.9%; p = 0. 001) more frequently than group 1 before TS2. For the sawtooth pattern, the likelihood ratio (LR) of a positive test was 2.70 and the LR of a negative test was 0.25, while for respiratory sounds it was 2.50 and 0.45, respectively. When the presence of a sawtooth pattern and of respiratory sounds was combined, the LR of a positive test rose to 14.7 and the LR of a negative test was 0.42. CONCLUSIONS: A sawtooth pattern and/or respiratory sounds over the trachea are good indicators of retained secretions in patients receiving MV and may indicate the need for TS. Conversely, the absence of a sawtooth pattern may rule out retained secretions. PMID- 11035684 TI - Acute respiratory failure in the United States: incidence and 31-day survival. AB - STUDY OBJECTIVES: To estimate the incidence of acute respiratory failure (ARF) in the United States and to analyze 31-day hospital mortality among a cohort of patients with ARF. DESIGN AND SETTING: Retrospective cohort drawn from the Nationwide Inpatient Sample of 6. 4 million discharges from 904 representative nonfederal hospitals during 1994. PATIENTS: All 61,223 patients in the sample whose discharge records indicated all of the following: acute respiratory distress or failure, mechanical ventilation, > or = 24 h of hospitalization, and age > or = 5 years. RESULTS: An estimated 329,766 patients discharged from nonfederal hospitals nationwide in 1994 met study criteria for ARF. The incidence of ARF was 137.1 hospitalizations per 100,000 US residents age > or = 5 years. Incidence increased nearly exponentially each decade until age 85 years. Overall, 35.9% of patients with ARF did not survive to hospital discharge. At 31 days, hospital mortality was 31.4%. According to the proportional hazards model, significant mortality hazards included age (> or = 80 years and > or = 30 years), multiorgan system failure (MOSF), HIV, chronic liver disease, and cancer. Hospital admission for coronary artery bypass, drug overdose, or trauma other than head injury or burns was associated with a reduced mortality hazard. Interaction was present between age and MOSF, trauma, and cancer. A point system derived from the hazard model classified patients into seven groups with distinct 31-day survival probabilities ranging from 24 to 99%. CONCLUSIONS: The incidence of ARF increases markedly with age and is especially high among persons > or = 65 years of age. Nonpulmonary hazards explain short-term (31-day) survival. PMID- 11035685 TI - A pilot study of penicillin skin testing in patients with a history of penicillin allergy admitted to a medical ICU. AB - BACKGROUND: Penicillin skin testing is an accurate method to determine whether a person with a history of penicillin allergy is at risk of having an immediate reaction to penicillin. A patient with a negative reaction to a skin test may be able to use a penicillin compound safely, which could reduce the use of broad spectrum antibiotics in this patient population. METHODS: We prospectively studied all patients with histories of penicillin allergy who were admitted to a medical ICU during a 3-month period and who received antibiotics. Skin testing was performed with benzylpenicilloyl polylysine and penicillin G. We determined the incidence of true allergy, the percentage of patients in whom antibiotic coverage was modified, and the safety of the test. RESULTS: Two hundred fifty seven patients were admitted to the medical ICU of The Cleveland Clinic Foundation during the study period. Twenty-four patients (9%), labeled as penicillin allergic and receiving antibiotics, were enrolled. Three patients (13%, 3 of 21) gave histories of type I reaction to penicillin and were not skin tested. Twenty patients (95%, 20 of 21) had negative skin test reactions to penicillin and positive skin test reactions to histamine control. One patient (4%, 1 of 21) with negative skin test reactions to both penicillin and histamine control had a test dose challenge with piperacillin that was well tolerated. There were no adverse events. Antibiotic coverage was changed in 10 patients (48%) as a result of skin testing. CONCLUSION: Most patients with histories of allergy to penicillin have negative reactions to skin tests and may receive penicillin safely. Penicillin skin testing can be utilized as a safe and effective strategy to reduce the use of broad-spectrum antibiotics. PMID- 11035686 TI - Performance of transport ventilator with patient-triggered ventilation. AB - OBJECTIVES: Transport ventilators with inspiratory triggering functions and pressure support-control modes have recently become commercially available. We evaluated these ventilators in comparison with a standard ICU ventilator. STUDY DESIGN: Laboratory study with a mechanical lung model. METHODS: We compared the performance of four transport ventilators (model 740, Mallinckrodt, Pleasanton, CA; TBird, Bird Products Corp, Palm Springs, CA; LTV1000, Pulmonetic Systems, Colton, CA; Esprit, Respironics, Vista, CA) with a standard ICU ventilator (model 7200ae; Mallinckrodt) using a test lung that simulated spontaneous breathing (compliance, 46.8 mL/cm H(2)O; resistance, 5 cm H(2)O/L/s). The settings of ventilators were positive end-expiratory pressure (PEEP) of 0 or 5 cm H(2)O, and pressure support (PS) of 0 or 10 cm H(2)O. The settings of the test lung were inspiratory time of 1 s, respiratory rate of 10/min, peak inspiratory flow of 40, 60, and 80 L/min. To evaluate inspiratory function at each setting, we measured the inspiratory delay time (DT), inspiratory trigger pressure (P-I), and the time for airway pressure to rise from the baseline pressure to 90% of the end inspiratory pressure (T(90%)); for expiratory function, supraplateau expiratory pressure (P-E) and the time constant (taue) for pressure decrease during exhalation were evaluated. Oxygen requirement was assessed as the time required to empty a 3.5-L oxygen tank. RESULTS: For inspiratory triggering, four transport ventilators had DT < 100 ms, which is considered clinically satisfactory, in all the settings except for PS 0 cm H(2)O, PEEP 0 cm H(2)O, and inspiratory flow of 80 L/min with LTV1000. P-I increased only in LTV1000 when PEEP was increased from 0 to 5 cm H(2)O. taue for the transport ventilators was > 50% shorter than for the ICU ventilator except for PS 0 cm H(2)O and PEEP 5 cm H(2)O with TBird. Oxygen requirement was lowest for the Esprit, followed by the 740, LTV1000, and TBird. CONCLUSION: The newer Food and Drug Administration-approved transport ventilators have performance indexes comparable to the ventilator currently used in ICUs and can probably be recommended for clinical use. PMID- 11035687 TI - Effects of cardiac glycosides on atrial contractile dysfunction after short-term atrial fibrillation. AB - BACKGROUND: Despite a long history of use in the treatment of paroxysmal atrial fibrillation (AF), the efficacy of cardiac glycosides has not been established. If such drugs are beneficial in this condition, the general view is that the benefit must be related to their inotropic actions. METHODS AND RESULTS: To assess the effects of the rapid-acting cardiac glycoside, acetylstrophanthidin (AS), on AF and AF-induced right atrial (RA) "stunning," RA wall motion (with ultrasonic crystals), RA pressure, and peak first derivative of pressure (dp/dt) (with microtip transducers) were measured before and after 5 min of high intensity rapid atrial stimulation (10 Hz; 10 mA; 1 ms) and after the cessation of poststimulation AF. Measurements were made in neurally intact and autonomically blockaded dogs both before and after the administration of AS (0.01 mg/kg IV bolus and 0.015 mg/kg/h IV infusion). AS prevented the post-AF reduction in RA peak dp/dt under neurally intact and autonomically blockaded conditions, and it prevented the post-AF increase in the RA end-systolic dimension and the decrease in the percentage of RA systolic shortening with autonomic blockade. AS was beneficial whether or not baseline inotropy was enhanced by AS. The duration of AF following atrial stimulation was the same before and after AS, but when compared to controls, AS treatment appeared to prolong AF. CONCLUSIONS: Cardiac glycosides exert a favorable effect on AF-induced RA stunning, but this action is unrelated to its effects on the duration of AF. PMID- 11035688 TI - Comparisons between sublingual and gastric tonometry during hemorrhagic shock. AB - STUDY OBJECTIVES: To compare sublingual tissue PCO(2), a disarmingly simple and noninvasive measurement of the severity of perfusion failure, with gastric tonometric PCO(2) during hemorrhagic shock in five male domestic pigs weighing between 35 and 40 kg. DESIGN: Prospective animal study. SETTING: Animal laboratory in a research institution. PARTICIPANTS: Domestic pigs. INTERVENTIONS: Hemorrhagic shock was induced by a modification of the Wigger's method. BP was maintained at 50 mm Hg for 120 min followed by reinfusion of shed blood at a rate of 100 mL/min with the aid of an infusion pump. MEASUREMENTS AND RESULTS: During bleeding, the mean arterial pressure decreased from an average of 127 to 42 mm Hg, and cardiac output decreased from 7.7 to 2.4 L/min. Arterial blood lactate concentration concurrently increased from 1.2 to 13.9 mmol/L. Sublingual PCO(2) (PslCO(2)) increased from 59 to 105 mm Hg, and gastric PCO(2) increased from 61 to 111 mm Hg. The correlation between time-coincident sublingual and gastric measurements of PCO(2) was r = 0.91 (p<0.0001). Bland-Altman analyses demonstrated a close correspondence between the two measurements. The reinfusion of shed blood promptly reversed the hemodynamic abnormalities and reestablished gastric and PslCO(2) to near baseline values. This contrasted with a delayed reversal of lactic acidosis. CONCLUSIONS: Under experimental conditions of hemorrhagic shock, sublingual capnometry yielded measurements that were interchangeable with those of gastric tonometry. PMID- 11035689 TI - HIV-Related pulmonary hypertension: analytic review of 131 cases. AB - OBJECTIVE: To report two new cases of HIV-related pulmonary hypertension and to review and analyze the existing reports on the subject. METHOD: Two new cases of HIV-related pulmonary hypertension are described, and the cases, case series, and related articles on the subject in all languages were identified through a comprehensive MEDLINE search. RESULTS: Among the 131 reviewed cases, 54% were male, and the age range was 2 to 56 years (mean, 33 years). The interval between the diagnosis of HIV disease and the diagnosis of pulmonary hypertension was 33 months. In 82% of cases, pulmonary hypertension was related solely to HIV infection. Presenting symptoms were progressive shortness of breath (85%), pedal edema (30%), nonproductive cough (19%), fatigue (13%), syncope or near-syncope (12%), and chest pain (7%). The mean (+/- SD) pulmonary arterial systolic BP was 67 +/- 18 mm Hg (n = 116), and diastolic BP was 40+/-11 mm Hg (n = 39). Pulmonary vascular resistance was 983+/-420 dyne. s. cm(-5) (n = 29). Chest radiographs demonstrated cardiomegaly (72%) and pulmonary artery prominence (71%). Right ventricular hypertrophy was the most common electrocardiographic finding (67%). Dilatation of the right heart chambers was the most common echocardiographic finding (98%). Plexogenic pulmonary arteriopathy was the most common histopathology (78%). Pulmonary function tests demonstrated mild restrictive patterns with variably reduced diffusing capacities. The responses to vasodilator agents and antiretroviral therapy was variable. Sixty-six patients died during a median follow-up period of 8 months. The median length of time from diagnosis to death was 6 months. CONCLUSION: HIV infection is an independent risk factor for the development of pulmonary hypertension. The appearance of unexplained cardiopulmonary symptoms in HIV-infected individuals should suggest pulmonary hypertension. PMID- 11035690 TI - The effect of inflammation on mucociliary clearance in asthma: an overview. AB - Mucociliary clearance (MCC) is one of the most important nonspecific defense mechanisms of the respiratory tract, and its impairment is a well-documented feature of chronic respiratory diseases, including asthma. In vitro and in vivo data suggest that several inflammatory mediators influence the mucociliary apparatus. Epithelial damage and functional abnormalities have been described in bronchial asthma, along with changes in mucus-secreting cells and the chemical and rheological properties of airway fluid. Although the mechanisms of MCC impairment in asthma are not clearly understood, data in the recent literature suggest that airway inflammation plays a major role. In this article, we review studies on MCC alterations in light of up-to-date findings on pathogenetic mechanisms in asthma. PMID- 11035691 TI - Chicken soup inhibits neutrophil chemotaxis in vitro. AB - Chicken soup has long been regarded as a remedy for symptomatic upper respiratory tract infections. As it is likely that the clinical similarity of the diverse infectious processes that can result in "colds" is due to a shared inflammatory response, an effect of chicken soup in mitigating inflammation could account for its attested benefits. To evaluate this, a traditional chicken soup was tested for its ability to inhibit neutrophil migration using the standard Boyden blindwell chemotaxis chamber assay with zymosan-activated serum and fMet-Leu-Phe as chemoattractants. Chicken soup significantly inhibited neutrophil migration and did so in a concentration-dependent manner. The activity was present in a nonparticulate component of the chicken soup. All of the vegetables present in the soup and the chicken individually had inhibitory activity, although only the chicken lacked cytotoxic activity. Interestingly, the complete soup also lacked cytotoxic activity. Commercial soups varied greatly in their inhibitory activity. The present study, therefore, suggests that chicken soup may contain a number of substances with beneficial medicinal activity. A mild anti-inflammatory effect could be one mechanism by which the soup could result in the mitigation of symptomatic upper respiratory tract infections. PMID- 11035692 TI - Medical and surgical treatment of parapneumonic effusions : an evidence-based guideline. AB - OBJECTIVE: A panel was convened by the Health and Science Policy Committee of the American College of Chest Physicians to develop a clinical practice guideline on the medical and surgical treatment of parapneumonic effusions (PPE) using evidence-based methods. OPTIONS AND OUTCOMES CONSIDERED: Based on consensus of clinical opinion, the expert panel developed an annotated table for evaluating the risk for poor outcome in patients with PPE. Estimates of the risk for poor outcome were based on the clinical judgment that, without adequate drainage of the pleural space, the patient with PPE would be likely to have any or all of the following: prolonged hospitalization, prolonged evidence of systemic toxicity, increased morbidity from any drainage procedure, increased risk for residual ventilatory impairment, increased risk for local spread of the inflammatory reaction, and increased mortality. Three variables, pleural space anatomy, pleural fluid bacteriology, and pleural fluid chemistry, were used in this annotated table to categorize patients into four separate risk levels for poor outcome: categories 1 (very low risk), 2 (low risk), 3 (moderate risk), and 4 (high risk). The panel's consensus opinion supported drainage for patients with moderate (category 3) or high (category 4) risk for a poor outcome, but not for patients with very low (category 1) or low (category 2) risk for a poor outcome. The medical literature was reviewed to evaluate the effectiveness of medical and surgical management approaches for patients with PPE at moderate or high risk for poor outcome. The panel grouped PPE management approaches into six categories: no drainage performed, therapeutic thoracentesis, tube thoracostomy, fibrinolytics, video-assisted thoracoscopic surgery (VATS), and surgery (including thoracotoiny with or without decortication and rib resection). The fibrinolytic approach required tube thoracostomy for administration of drug, and VATS included post procedure tube thoracostomy. Surgery may have included concomitant lung resection and always included postoperative tube thoracostomy. All management approaches included appropriate treatment of the underlying pneumonia, including systemic antibiotics. Criteria for including articles in the panel review were adequate data provided for >/=20 adult patients with PPE to allow evaluation of at least one relevant outcome (death or need for a second intervention to manage the PPE); reasonable assurance provided that drainage was clinically appropriate (patients receiving drainage were either category 3 or category 4) and drainage procedure was adequately described; and original data were presented. The strength of panel recommendations on management of PPE was based on the following approach: level A, randomized, controlled trials with consistent results or individual randomized, controlled trial with narrow confidence interval (CI); level B, controlled cohort and case control series; level C, historically controlled series and case series; and level D, expert opinion without explicit critical appraisal or based on physiology, bench research, or "first principles." EVIDENCE: The literature review revealed 24 articles eligible for full review by the panel, 19 of which dealt with the primary management approach to PPE and 5 with a rescue approach after a previous approach had failed. Of the 19 involving the primary management approach to PPE, there were 3 randomized, controlled trials, 2 historically controlled series, and 14 case series. The number of patients included in the randomized controlled trials was small; methodologic weaknesses were found in the 19 articles describing the results of primary management approaches to PPE. The proportion and 95% CI of patients suffering each of the two relevant outcomes (death and need for a second intervention to manage the PPE) were calculated for the pooled data for each management approach from the 19 articles on the primary management approach. (ABST PMID- 11035693 TI - Ethical decision making and patient autonomy: a comparison of physicians and patients in Japan and the United States. AB - BACKGROUND: Patient-centered decision making, which in the United States is typically considered to be appropriate, may not be universally endorsed, thereby harboring the potential to complicate the care of patients from other cultural backgrounds in potentially unrecognized ways. This study compares the attitudes toward ethical decision making and autonomy issues among academic and community physicians and patients of medical center outpatient clinics in Japan and the United States. METHODS: A questionnaire requesting judgments about seven clinical vignettes was distributed (in English or Japanese) to sample groups of Japanese physicians (n = 400) and patients (n = 65) as well as US physicians (n = 120) and patients (n = 60) that were selected randomly from academic institutions and community settings in Japan (Tokyo and the surrounding area) and the United States (the Stanford/Palo Alto, CA, area). Responses were obtained from 273 Japanese physicians (68%), 58 Japanese patients (89%), 98 US physicians (82%), and 55 US patients (92%). Physician and patient sample groups were compared on individual items, and composite scores were derived from subsets of items relevant to patient autonomy, family authority, and physician authority. RESULTS: A majority of both US physicians and patients, but only a minority of Japanese physicians and patients, agreed that a patient should be informed of an incurable cancer diagnosis before their family is informed and that a terminally ill patient wishing to die immediately should not be ventilated, even if both the doctor and the patient's family want the patient ventilated (Japanese physicians and patients vs US physicians and patients, p < 0.001). A majority of respondents in both Japanese sample groups, but only a minority in both US sample groups, agreed that a patient's family should be informed of an incurable cancer diagnosis before the patient is informed and that the family of an HIV-positive patient should be informed of this disease status despite the patient's opposition to such disclosure (Japanese physicians and patients vs US physicians and patients, p < 0.001). Physicians in both Japan and the United States were less likely than patients in their respective countries to agree with physician assistance in the suicide of a terminally ill patient (Japanese physicians and patients vs US physicians and patients, p < 0.05). Across various clinical scenarios, all four respondent groups accorded greatest authority to the patient, less to the family, and still less to the physician when the views of these persons conflicted. Japanese physicians and patients, however, relied more on family and physician authority and placed less emphasis on patient autonomy than the US physicians and patients sampled. Younger respondents placed less emphasis on family and physician authority. CONCLUSIONS: Family and physician opinions are accorded a larger role in clinical decision making by the Japanese physicians and patients sampled than by those in the United States, although both cultures place a greater emphasis on patient preferences than on the preferences of the family or physician. Our results are consistent with the view that cultural context shapes the relationship of the patient, the physician, and the patient's family in medical decision making. The results emphasize the need for clinicians to be aware of these issues that may affect patient and family responses in different clinical situations, potentially affecting patient satisfaction and compliance with therapy. PMID- 11035694 TI - Asthma-like symptoms in wood product plant workers exposed to methylene diphenyl diisocyanate. AB - BACKGROUND: Diisocyanates, a group of highly reactive chemicals, have frequently been associated with occupational asthma. We evaluated respiratory health in workers at a new wood products manufacturing plant that uses methylene diphenyl diisocyanate (MDI), and was designed and operated with a goal of minimizing worker exposures. METHODS: Health surveys using standardized respiratory questionnaires were done prior to the initial use of diisocyanates in the plant, and semiannually thereafter for a period of 2 years. Other testing included occupational and work practice histories, serial peak flow measurements, spirometry, methacholine challenge, and measurement of specific IgE antibodies to MDI-albumin conjugate. RESULTS: Of 214 plant employees who participated in at least one health survey, a follow-up survey was also available from 178 employees (83%). New-onset asthma-like symptoms (NAS) were reported by 15 of 56 workers (27%) in areas with the highest potential for exposures to liquid MDI monomer and prepolymer, vs 0 of 43 workers in the lowest potential exposure areas (p = 0.001). In the areas with high potential exposure, NAS developed in 47% of workers who had noted MDI skin staining, vs 19% without skin stains (p = 0.07). Working around and cleaning up liquid MDI represented a significant risk for asthma-like symptoms in both current smokers and nonsmokers; work with finished wood products did not. Asthma-like symptoms were associated with variable airflow limitation (odds ratio [OR], 5.0; confidence interval [CI], 1.4 to 18.7) and specific IgE to MDI-albumin (OR, 3.2; CI, 1.1 to 9.0), but not with skin prick tests to common aeroallergens (OR, 1.1; CI, 0.5 to 2.7). CONCLUSIONS: During the first 2 years of operation, in a plant designed and operated to control exposure to diisocyanates, the development of asthma-like symptoms was reported in a relatively high proportion of the employees who worked with liquid MDI. To prevent asthma symptoms among workers, careful control of respiratory tract exposures associated with liquid MDI is important, especially during cleanup activities. Strict limitation of skin contact with diisocyanates may also be necessary. PMID- 11035695 TI - A 58-year old woman with recurrent productive cough and diarrhea. PMID- 11035696 TI - 54-year-old man with dyspnea, cough, and hypoxemia. PMID- 11035697 TI - A first event of dyspnea in an infant. PMID- 11035698 TI - Catamenial hemoptysis from tracheobronchial endometriosis: reappraisal of diagnostic value of bronchoscopy and bronchial brush cytology. AB - OBJECTIVES: To analyze the clinical data of four patients with a diagnosis of tracheobronchial endometriosis, and to reappraise the diagnostic value of bronchoscopy and bronchial brush cytology in these patients. METHODS: We conducted a retrospective study of four patients with documented tracheobronchial endometriosis treated at National Taiwan University Hospital from 1994 to 1998. The complete histories, diagnostic time interval, results of physical examinations, laboratory data, bronchoscopic findings, cytologic results, chest radiographs, and chest CT of these patients were analyzed. RESULTS: These patients tend to be younger and nonmultiparous as compared to other patients with thoracic endometriosis. Bronchoscopic examination performed within 1 day or 2 days of menses disclosed multiple purplish-red submucosal patches bilaterally that bled easily when touched. Cytologic evaluation of the brushing specimens demonstrated clusters of small cuboid cells consistent with an endometrial origin. Follow-up bronchoscopic examination in the middle of the menstrual cycle showed disappearance of the previous tracheobronchial lesions. The mean diagnostic interval was 3.25 months. All four patients were successfully treated with danazol therapy. CONCLUSIONS: Tracheobronchial endometriosis consists of a special subgroup of patients with thoracic endometriosis. Proper timing of bronchoscopic examination plays an important diagnostic role in these patients. Cytologic features as well as cyclic changes in bronchoscopic findings are sufficient to warrant the diagnosis. The results of treatment with danazol in these patients seemed favorable. PMID- 11035699 TI - Methemoglobinemia after infusion of ifosfamide chemotherapy: first report of a potentially serious adverse reaction related to ifosfamide. AB - Acute formation of methemoglobin is a life-threatening condition caused by multiple medications. In this article we report the first case of methemoglobinemia in a patient with metastatic uterine leiomyosarcoma, after infusion of ifosfamide chemotherapy. The patient recovered after prompt diagnosis and treatment of the condition. A mechanism for the formation of methemoglobin as a result of the ifosfamide infusion is offered. PMID- 11035700 TI - Successful treatment of juvenile laryngeal papillomatosis-related multicystic lung disease with cidofovir: case report and review of the literature. AB - Cidofovir, a nucleoside analog antiviral agent, has been used with moderate success in the treatment of juvenile laryngeal papillomatosis (JLP) by direct intralesional injection. We report the first case where IV cidofovir was used successfully to treat a rare but lethal multicystic lung disease complicating JLP. A 35-year-old woman with a history of JLP requiring multiple laser ablations of laryngeal papillomata each year presented with hemoptysis and was found on CT scan to have bilateral, multiple pulmonary nodules and cysts. The results of BAL fluid analysis demonstrated no evidence of malignancy, and cultures were negative for fungi and mycobacteria. Molecular DNA typing of a biopsy specimen obtained from a laryngeal papilloma confirmed infection with human papilloma virus type 11. She received 12 months of treatment with IV cidofovir followed by 9 months of combined treatmentwith IV cidofovir and subcutaneous interferon-alpha-2A. This therapeutic regime resulted in a markedly decreased requirement for surgical removal of laryngeal papillomata, and CT scanning documented the regression of the lesions in the lung parenchyma that persisted after the discontinuation of therapy. The results of this case demonstrate that cidofovir may be used successfully to treat JLP-related lung disease and suggest that further studies are warranted. PMID- 11035701 TI - Gastroesophageal reflux as a reversible cause of allograft dysfunction after lung transplantation. AB - Gastroesophageal reflux (GER) is increasingly recognized as contributing to a number of pulmonary disorders. The relationship of GER to pulmonary allograft dysfunction after lung transplantation is unknown. In this report, we describe a lung transplant recipient who developed an acute decline in pulmonary function several months after a retransplantation for chronic rejection. A pulmonary workup at that time, including bronchoscopy with biopsy, revealed bronchial inflammation with no allograft rejection or infection. Because of increasing GI symptoms after retransplantation, the patient also underwent additional testing, which revealed severe acid reflux. The treatment of this patient's acid reflux with Nissen fundoplication surgery resulted in a prompt and sustained improvement in his pulmonary function. We suggest that GER should be considered among the potential causes of allograft dysfunction after lung transplantation. PMID- 11035702 TI - Cardiac rhabdomyoma in an adult patient presenting with ventricular arrhythmia. AB - Cardiac rhabdomyomas are extremely uncommon in the adult patient. We describe a previously healthy man who presented with ventricular arrhythmias resulting from a right ventricular, cardiac rhabdomyoma. Echocardiography, CT scanning, and MRI are recognized as useful diagnostic modalities for intracardiac lesions. Cardiac catheterization in our patient demonstrated the presence of a tumor blush. This has not previously been reported with cardiac rhabdomyomas. Although lesions may spontaneously regress, surgery is often necessary and frequently resolves the underlying arrhythmia. PMID- 11035703 TI - Pheochromocytoma crisis, cardiomyopathy, and hemodynamic collapse. AB - Pheochromocytoma is a notorious clinical entity. Although suspicion is aroused by severe hypertension in young patients, this sign is often absent. We present a case in which early absence of hypertension and nonspecific signs and symptoms led to failure of prompt diagnosis. The delay proved fatal when the patient developed fulminant pheochromocytoma crisis. This case illustrates a variety of clinical features seen from the vantage of the evolution of the disease as it went unrecognized. The patient's course underscores the importance of familiarity with the gamut of manifestations for timely diagnosis, and the priority of the latter given the looming risk of overwhelming complications. PMID- 11035704 TI - Primary pulmonary hypertension and thyroid disease. PMID- 11035705 TI - Prognostic value of von Willebrand factor concentrations in pulmonary hypertension. PMID- 11035706 TI - Ciprofloxacin vs. the pneumococcus. PMID- 11035707 TI - Analysis of results in posttransplant lymphoproliferative disorder. PMID- 11035708 TI - Follow-up in Montelukast treatment. PMID- 11035709 TI - Epoprostenol for treatment of pulmonary hypertension in patients with systemic lupus erythematosus. PMID- 11035710 TI - Genetic manipulation of rickettsiae: a preview. PMID- 11035711 TI - Secretion of RTX leukotoxin by Actinobacillus actinomycetemcomitans. AB - Actinobacillus actinomycetemcomitans, the etiologic agent for localized juvenile periodontitis and certain other human infections, such as endocarditis, expresses a leukotoxin that acts on polymorphonuclear leukocytes and macrophages. Leukotoxin is a member of the highly conserved repeat toxin (RTX) family of bacterial toxins expressed by a variety of pathogenic bacteria. While the RTX toxins of other bacterial species are secreted, the leukotoxin of A. actinomycetemcomitans is thought to remain associated with the bacterial cell. We have examined leukotoxin production and localization in rough (adherent) and smooth (nonadherent) strains of A. actinomycetemcomitans. We found that leukotoxin expressed by the rough, adherent, clinical isolate CU1000N is indeed cell associated, as expected. However, we were surprised to find that smooth, nonadherent strains of A. actinomycetemcomitans, including Y4, JP2 (a strain expressing a high level of toxin), and CU1060N (an isogenic smooth variant of CU1000N), secrete an abundance of leukotoxin into the culture supernatants during early stages of growth. After longer times of incubation, leukotoxin disappears from the supernatants, and its loss is accompanied by the appearance of a number of low-molecular-weight polypeptides. The secreted leukotoxin is active, as evidenced by its ability to kill HL-60 cells in vitro. We found that the growth phase and initial pH of the growth medium significantly affect the abundance of secreted leukotoxin, and we have developed a rapid (<2 h) method to partially purify large amounts of leukotoxin. Remarkably, mutations in the tad genes, which are required for tight nonspecific adherence of A. actinomycetemcomitans to surfaces, cause leukotoxin to be released from the bacterial cell. These studies show that A. actinomycetemcomitans has the potential to secrete abundant leukotoxin. It is therefore appropriate to consider a possible role for leukotoxin secretion in the pathogenesis of A. actinomycetemcomitans. PMID- 11035712 TI - NO contributes to proliferative suppression in a murine model of filariasis. AB - Infection of BALB/c mice with microfilariae (mf) of Brugia pahangi leads to the suppression of antigen (Ag)-specific proliferative responses in the spleen. The proliferative defect is dependent on inducible nitric oxide synthase (iNOS) activity, since inhibition of iNOS with either L-N-monomethyl arginine (L-NMMA) or aminoguanidine reversed defective proliferation. Splenocytes from mf-infected animals produce high levels of gamma interferon (IFN-gamma) upon in vitro restimulation with Ag, and experiments in IFN-gamma receptor-deficient (IFN-gamma R(-/-)) mice demonstrated that signaling via the IFN-gamma R is essential in the induction of NO production and subsequent proliferative suppression. Restimulation of splenocytes from mf-infected animals with an extract of Acanthocheilonema viteae, a related filarial worm which lacks endosymbiotic bacteria, also resulted in NO production and proliferative suppression, demonstrating that lipopolysaccharide of bacterial origin is not essential to the induction of iNOS activity. These results extend previous observations that infection with different life cycle stages of Brugia leads to the development of differentially polarized immune responses and demonstrate one method by which these differences may exert their effects on the proliferative potential of cells from infected animals. PMID- 11035713 TI - Chemokine C10 promotes disease resolution and survival in an experimental model of bacterial sepsis. AB - Previous studies have suggested that the C-C chemokine C10 is involved in the chronic stages of host defense reactions. The present study addressed the role of C10 in a murine model of septic peritonitis, induced by cecal ligation and puncture (CLP). Unlike other C-C chemokines, C10 levels in the peritoneal wash were increased approximately 30-fold above baseline levels at 48 h after CLP surgery. Immunoneutralization of peritoneal C10 levels with polyclonal anti-C10 antiserum during CLP-induced peritonitis negatively impacted mouse survival over 4 days. In contrast, when 500 ng of recombinant murine C10 was administered immediately after CLP surgery, the 4-day survival rate increased from 20% to over 60%. The C10 therapy appeared to facilitate a rapid and significant enhancement of the levels of tumor necrosis factor alpha (TNF-alpha) and monocyte chemoattractant protein-1 (MCP-1) and a later increase in interleukin-13 (IL-13) levels in the peritoneal cavity. In vitro studies showed that the combination of IL-1beta and C10 markedly augmented TNF-alpha synthesis by peritoneal macrophages and that C10 synthesis was induced in these cells following their exposure to IL 13. At 24 h after CLP surgery, only 25% of C10-treated mice were bacteremic versus 85% of the control group that exhibited dissemination of bacteria into the circulation. The lack of bacteremia in C10-treated mice appeared to be related, in part, to in vitro evidence that C10 significantly enhanced the bacterial phagocytic activity of peritoneal macrophages. In addition, in vivo evidence suggested that C10 therapy significantly reduced the amount of material that leaked from the damaged gut. Taken together, the results of this study demonstrate that the C10 chemokine rapidly promotes disease resolution in the CLP model through its direct effects on the cellular events critically involved in host defense during septic peritonitis. PMID- 11035714 TI - The locus of enterocyte effacement (LEE)-encoded regulator controls expression of both LEE- and non-LEE-encoded virulence factors in enteropathogenic and enterohemorrhagic Escherichia coli. AB - Regulation of virulence gene expression in enteropathogenic Escherichia coli (EPEC) and enterohemorrhagic E. coli (EHEC) is incompletely understood. In EPEC, the plasmid-encoded regulator Per is required for maximal expression of proteins encoded on the locus of enterocyte effacement (LEE), and a LEE-encoded regulator (Ler) is part of the Per-mediated regulatory cascade upregulating the LEE2, LEE3, and LEE4 promoters. We now report that Ler is essential for the expression of multiple LEE-located genes in both EPEC and EHEC, including those encoding the type III secretion pathway, the secreted Esp proteins, Tir, and intimin. Ler is therefore central to the process of attaching and effacing (AE) lesion formation. Ler also regulates the expression of LEE-located genes not required for AE-lesion formation, including rorf2, orf10, rorf10, orf19, and espF, indicating that Ler regulates additional virulence properties. In addition, Ler regulates the expression of proteins encoded outside the LEE that are not essential for AE lesion formation, including TagA in EHEC and EspC in EPEC. delta ler mutants of both EPEC and EHEC show altered adherence to epithelial cells and express novel fimbriae. Ler is therefore a global regulator of virulence gene expression in EPEC and EHEC. PMID- 11035715 TI - Early nonspecific immune responses and immunity to blood-stage nonlethal Plasmodium yoelii malaria. AB - The early role of natural killer cells and gamma delta T cells in the development of protective immunity to the blood stage of nonlethal Plasmodium yoelii infection was studied. Splenic cytokine levels were measured 24 h after infection of natural killer cell-depleted immunodeficient and littermate mice or transiently T-cell-depleted normal mice. Splenic gamma interferon levels were significantly increased above background in immunodeficient and littermate mice 24 h after infection. Depletion of natural killer cells resulted in markedly depressed gamma interferon levels and poor control of parasitemia, particularly in severe combined immunodeficient mice. In the littermates, gamma interferon levels were partially reduced, but parasitemias were resolved normally. However, in athymic mice, natural killer cell depletion had no effect on gamma interferon production. Levels of tumor necrosis factor alpha were increased in all animals 24 h after infection, and responses were not affected by natural killer cell depletion. However, in T-cell-depleted animals, both gamma interferon and tumor necrosis factor alpha levels were decreased 24 h after infection, and depleted mice were unable to control their parasitemia. These results suggest that the early production of both cytokines is important in the early control of parasitemia and that both natural killer and gamma delta T cells contribute equally towards their production. The data also suggest that the subsequent resolution of infection requires early production of gamma interferon, which might act by switching on the appropriate T-helper-cell subsets and other essential parasitotoxic effector mechanisms. PMID- 11035716 TI - Antigenic variation of Anaplasma marginale by expression of MSP2 mosaics. AB - Anaplasma marginale is a tick-borne pathogen, one of several closely related ehrlichial organisms that cause disease in animals and humans. These Ehrlichia species have complex life cycles that require, in addition to replication and development within the tick vector, evasion of the immune system in order to persist in the mammalian reservoir host. This complexity requires efficient use of the small ehrlichial genome. A. marginale and related ehrlichiae express immunoprotective, variable outer membrane proteins that have similar structures and are encoded by polymorphic multigene families. We show here that the major outer membrane protein of A. marginale, MSP2, is encoded on a polycistronic mRNA. The genomic expression site for this mRNA is polymorphic and encodes numerous amino acid sequence variants in bloodstream populations of A. marginale. A potential mechanism for persistence is segmental gene conversion of the expression site to link hypervariable msp2 sequences to the promoter and polycistron. PMID- 11035717 TI - Genetic and functional analysis of a PmrA-PmrB-regulated locus necessary for lipopolysaccharide modification, antimicrobial peptide resistance, and oral virulence of Salmonella enterica serovar typhimurium. AB - The two-component regulatory system PmrA-PmrB confers resistance of Salmonella spp. to cationic antimicrobial peptides (AP) such as polymyxin (PM), bactericidal/permeability-increasing protein, and azurocidin. This resistance occurs by transcriptional activation of two loci termed pmrE and pmrHFIJKLM. Both pmrE and pmrHFIJKLM produce products required for the biosynthesis of lipid A with 4-aminoarabinose (Ara4N). Ara4N addition creates a more positively charged lipopolysaccharide (LPS) and thus reduces cationic AP binding. Experiments were conducted to further analyze the regulation of the pmrHFIJKLM operon and the role of this operon and the surrounding genomic region in LPS modification and antimicrobial peptide resistance. The pmrHFIJKLM genes are cotranscribed and over 3,000-fold regulated by PmrA-PmrB. The pmrHFIJKLM promoter bound PmrA, as determined by gel shift analysis, as did a 40-bp region of the PmrA-PmrB regulated pmrCAB promoter. Construction of nonpolar mutations in the pmrHFIJKLM genes showed that all except pmrM were necessary for the Ara4N addition to lipid A and PM resistance. The flanking genes of the operon (pmrG and pmrD) were not necessary for PM resistance, but pmrD was shown to be regulated by the PhoP-PhoQ regulatory system. BALB/c mice inoculated with pmrA and pmrHFIJKLM mutant strains demonstrated virulence attenuation when the strains were administered orally but not when they were administered intraperitoneally, indicating that Ara4N addition may be important for resistance to host innate defenses within intestinal tissues. PMID- 11035718 TI - Phagocytosis and protein processing are required for presentation of Cryptococcus neoformans mitogen to T lymphocytes. AB - In addition to eliciting antigen specific T-cell-mediated immunity, Cryptococcus neoformans possesses a mitogen (CnM) that activates naive T cells to proliferate. This mechanism of T-cell activation is accessory cell dependent and major histocompatibility complex unrestricted. CnM-induced T-cell proliferation correlates with internalization of the organism, suggesting that intracellular processing is required to liberate CnM prior to presentation to T cells. To determine whether phagocytosis and processing are required, various inhibitors of accessory cell uptake and processing were used. C. neoformans was observed within the accessory cells. Paraformaldehyde fixation of the accessory cell abrogated presentation of CnM to T cells, indicating that a dynamic accessory cell surface was required. A lysosomotropic agent abrogated the response to CnM but had no effect on a control stimulus that did not require processing. Both aspartic acid and cysteine protease inhibitors blocked effective processing of CnM, so that it was unable to stimulate T cells. Finally, an inhibitor of microfilament polymerization abrogated proliferation to CnM. These results indicate that the mitogenic activity of C. neoformans requires phagocytosis of the organism, lysosomal or endosomal processing, proteolytic activity, and microfilament polymerization and intracellular transport as a prerequisite for T-cell proliferation. PMID- 11035720 TI - Role of neutrophil leukocytes in cutaneous infection caused by Staphylococcus aureus. AB - Despite the high prevalence of cutaneous infections, little is known about the role of host immune responsiveness during Staphylococcus aureus dermatitis. We have recently described a murine model of infectious dermatitis induced by superantigen-producing S. aureus. To assess the role of neutrophils in staphylococcal dermatitis, mice were given granulocyte-depleting monoclonal antibody prior to and on several occasions following intracutaneous inoculation with staphylococci. The granulocyte-depleted mice that had been intradermally inoculated with S. aureus developed crusted ulcerations which tended not to heal, whereas animals injected with control monoclonal antibody displayed only minor and transient skin lesions. The finding of severe ulcerations in neutropenic mice correlated with a significantly higher burden of bacteria in the blood and skin during the early phase of the infection. Importantly, while mice with an intact granulocyte population showed only limited skin infection, bacteremia occurred in the great majority of the neutrophil-depleted animals. As a consequence, the latter individuals exhibited significantly increased levels of the proinflammatory cytokine interleukin-6 and specific antibodies to staphylococcal cell wall components and toxic shock syndrome toxin-1 in the serum. Our data point to a crucial protective role of granulocytes in S. aureus dermatitis. PMID- 11035719 TI - Modulation of B-cell proliferative response by a soluble extract of Nippostrongylus brasiliensis. AB - We and others have previously shown that nematodes or nematode products can stimulate or inhibit the generation of lymphocyte responses, suggesting that nematodes exert diverse effects on the developing immune responses of their host. In this study we examined the immunomodulatory effect of a soluble extract of Nippostrongylus brasiliensis (adult worm homogenate [AWH]) on B-cell responsiveness. We found that the extract inhibited the proliferation of B cells to lipopolysaccharide (LPS) stimulation in a dose-dependent manner. This effect was specific to B cells, since the extract did not inhibit T-cell proliferation to concanavalin A or anti-CD3 stimulation. The data presented here confirm that the extract is not toxic to B cells. We present evidence that the active factor is proteinaceous in nature and that the inhibitory activity is restricted to the adult stage of Nb. The extract does not appear to interfere with early activation events since it can be added up to 48 h after LPS stimulation, and it inhibited responses to phorbol myristate acetate and ionomycin. Furthermore, the proliferation of B cells to other activators was also inhibited by AWH. This observation shows that the inhibitory activity of AWH is not restricted to LPS mediated B-cell proliferation. We present evidence that, in the absence of accessory cells, the inhibitory effect of the extract was ablated. This observation shows that the activity of AWH is not mediated directly on B cells but is mediated via the production of negative signals from accessory cells (macrophages), which affect a downstream pathway required by all B-cell activators tested. These effects on B-cell and accessory cell function are likely to have a significant effect on the outcome of infections experienced concurrently. PMID- 11035721 TI - Detection and characterization of autoagglutination activity by Campylobacter jejuni. AB - In several gram-negative bacterial pathogens, autoagglutination (AAG) activity is a marker for interaction with host cells and virulence. Campylobacter jejuni strains also show AAG, but this property varies considerably among strains. To examine the characteristics of C. jejuni AAG, we developed a quantitative in vitro assay. For strain 81-176, which shows high AAG, activity was optimal for cells grown for < or = 24 h, was independent of growth temperature, and was best measured for cells suspended in phosphate-buffered saline at 25 degrees C for 24 h. AAG activity was heat labile and was abolished by pronase or acid-glycine (pH 2.2) treatment but not by lipase, DNase, or sodium metaperiodate. Strain 4182 has low AAG activity, but extraction with water increased AAG, suggesting the loss of an inhibitor. Strain 6960 has weak AAG with no effect due to water extraction. Our study with clinical isolates suggests that C. jejuni strains may be grouped into three AAG phenotypes. A variant derived from strain 81116 that is flagellate but immotile showed the strong AAG exhibited by the parent strain, suggesting that motility per se is not necessary for the AAG activity. AAG correlated with both bacterial hydrophobicity and adherence to INT407 cells. Mutants which lack flagella (flaA, flaB, and flbA) or common cell surface antigen (peb1A) were constructed in strain 81-176 by natural transformation-mediated allelic exchange. Both AAG activity and bacterial hydrophobicity were abolished in the aflagellate mutants but not the peb1A mutant. In total, these findings indicate that C. jejuni AAG is highly associated with flagellar expression. PMID- 11035722 TI - Bacteroides fragilis NCTC9343 produces at least three distinct capsular polysaccharides: cloning, characterization, and reassignment of polysaccharide B and C biosynthesis loci. AB - Bacteroides fragilis produces a capsular polysaccharide complex (CPC) that is directly involved in its ability to induce abscesses. Two distinct capsular polysaccharides, polysaccharide A (PS A) and PS B, have been shown to be synthesized by the prototype strain for the study of abscesses, NCTC9343. Both of these polysaccharides in purified form induce abscesses in animal models. In this study, we demonstrate that the CPC of NCTC9343 is composed of at least three distinct capsular polysaccharides: PS A, PS B, and PS C. A previously described locus contains genes whose products are involved in the biosynthesis of PS C rather than PS B as was originally suggested. The actual PS B biosynthesis locus was cloned, sequenced, and found to contain 22 genes in an operon-type structure. A mutant with a large chromosomal deletion of the PS B biosynthesis locus was created so that the contribution of PS B to the formation of abscesses could be assessed in a rodent model. Although purified PS B can induce abscesses, removal of this polysaccharide does not attenuate the organism's ability to induce abscesses. PMID- 11035723 TI - Genetic diversity of the capsular polysaccharide C biosynthesis region of Bacteroides fragilis. AB - A genetic approach was used to assess the heterogeneity of the capsular polysaccharide C (PS C) biosynthesis locus of Bacteroides fragilis and to determine whether distinct loci contain genes whose products are likely to be involved in conferring charged groups that enable the B. fragilis capsular polysaccharides to induce abscesses. A collection of 50 B. fragilis strains was examined. PCR analysis demonstrated that the genes flanking the PS C biosynthesis region are conserved, whereas the genes within the loci are heterogeneous. Only cfiA(+) B. fragilis strains, which represent 3% of the clinical isolates of B. fragilis, displayed heterogeneity in the regions flanking the polysaccharide biosynthesis genes. Primers were designed in the conserved regions upstream and downstream of the PS C locus and were used to amplify the region from 45 of the 50 B. fragilis strains studied. Fourteen PS C genetic loci could be differentiated by a combination of PCR and extended PCR. These loci ranged in size from 14 to 26 kb. Hybridization analysis with genes from the PS C loci of strains 9343 and 638R revealed that the majority of strains contain homologs of wcgC (N-acetylmannosamine dehydrogenase), wcfF (putative dehydrogenase), and wcgP (putative aminotransferase). The data suggest that the synthesis of polysaccharides that have zwitterionic characteristics rendering them able to induce abscesses is common in B. fragilis. PMID- 11035724 TI - A protective glycosylphosphatidylinositol-anchored membrane protein of Plasmodium yoelii trophozoites and merozoites contains two epidermal growth factor-like domains. AB - Using sera from mice immunized and protected against Plasmodium yoelii malaria, we identified a novel blood-stage antigen gene, pypag-2. The 2.1-kb pypag-2 cDNA contains a single open reading frame that encodes a 409-amino-acid protein with a predicted molecular mass of 46.8 kDa. Unlike many characterized plasmodial antigens, blocks of tandemly repeated amino acids are lacking in the pypAg-2 protein sequence. Recombinant pypAg-2, comprising the full-length protein minus the predicted N-terminal signal and C-terminal anchor sequences, was produced and used to raise a high-titer polyclonal rabbit antiserum. This antiserum was used to identify and characterize the native protein through immunoblotting, immunoprecipitation and immunofluorescence assays. Consistent with the presence of a glycosylphosphatidylinositol anchor, pypAg-2 fractionated with the detergent phase of Triton X-114-solubilized proteins and could be metabolically labeled with [(3)H]palmitic acid. By immunofluorescence, pypAg-2 expression was localized to both the trophozoite and merozoite membranes. Similar to Plasmodium falciparum merozoite surface protein 1, pypAg-2 contains two C-terminal epidermal growth factor (EGF)-like domains. Most importantly, immunization with recombinant pypAg 2 protected mice against lethal P. yoelii malaria. Thus, pypAg-2 is a target of protective immune responses and represents a novel addition to the family of merozoite surface proteins that contain one or more EGF-like domains. PMID- 11035725 TI - The Cryptococcus neoformans gene DHA1 encodes an antigen that elicits a delayed type hypersensitivity reaction in immune mice. AB - When mice are vaccinated with a culture filtrate from Cryptococcus neoformans (CneF), they mount a protective cell-mediated immune response as detected by dermal delayed-type hypersensitivity (DTH) to CneF. We have identified a gene (DHA1) whose product accounts at least in part for the DTH reactivity. Using an acapsular mutant (Cap-67) of C. neoformans strain B3501, we prepared a culture filtrate (CneF-Cap67) similar to that used for preparing the commonly used skin test antigen made with C. neoformans 184A (CneF-184A). CneF-Cap67 elicited DTH in mice immunized with CneF-184A. Deglycosylation of CneF-Cap67 did not diminish its DTH activity. Furthermore, size separation by either chromatography or differential centrifugation identified the major DTH activity of CneF-Cap67 to be present in fractions that contained proteins of approximately 19 to 20 kDa. Using N-terminal and internal amino acid sequences derived from the 20-kDa band, oligonucleotide primers were designed, two of which produced a 776-bp amplimer by reverse transcription-PCR (RT-PCR) using RNA from Cap-67 to prepare cDNA for the template. The amplimer was used as a probe to isolate clones containing the full length DHA1 gene from a phage genomic library prepared from strain B3501. The full-length cDNA was obtained by 5' rapid amplification of cDNA ends and RT-PCR. Analysis of DHA1 revealed a similarity between the deduced open reading frame and that of a developmentally regulated gene from Lentinus edodes (shiitake mushroom) associated with fruiting-body formation. Also, the gene product contained several amino acid sequences identical to those determined biochemically from the purified 20-kDa peptide encoded by DHA1. Recombinant DHA1 protein expressed in Escherichia coli was shown to elicit DTH reactions similar to those elicited by CneF-Cap67 in mice immunized against C. neoformans. Thus, DHA1 is the first gene to be cloned from C. neoformans whose product has been shown to possess immunologic activity. PMID- 11035726 TI - Preparation and preclinical evaluation of a novel liposomal complete-core lipopolysaccharide vaccine. AB - Our objective is to develop a prophylactic vaccine strategy that can be evaluated for surgical and other high-risk hospitalized patients. In this paper, we describe the preparation and preclinical evaluation of a liposomal complete-core lipopolysaccharide (LPS) vaccine that is nontoxic and broadly antigenic. Complete core (Ra-chemotype) LPSs were isolated from four gram-negative bacterial strains (Escherichia coli K-12, E. coli R1, Pseudomonas aeruginosa PAC608, and Bacteroides fragilis), mixed together to form a cocktail of complete-core LPSs, and then incorporated into multilamellar liposomes consisting of dimyristoyl phosphatidyl choline, dimyristoyl phosphatidylglycerol, and cholesterol in a 4:1:4 molar ratio. The endotoxic activities of these LPS-containing liposomes were less than 0.1% of the endotoxicities of the original free LPSs as measured by the Limulus amoebocyte lysate assay. In vivo administration of liposomal complete-core LPS mixed with Al(OH)(3) to rabbits resulted in no pyrogenicity or overt toxicity over a 7-day period. In immunoblots, sera from rabbits following active immunization elicited cross-reactive antibodies to a large panel of rough and smooth LPSs from numerous clinically relevant gram-negative bacteria, including E. coli (serotypes O1, O4, O6, O8, O12, O15, O18, O75, O86, O157, and O111), P. aeruginosa (Fisher-Devlin serotypes 1, 2, and 3, which correspond to International Antigenic Typing Scheme types 6, 11, and 2, respectively), Klebsiella pneumoniae (serotypes O1, O2ab, and O3), B. fragilis, and Bacteroides vulgatus. Active immunization of mice with liposomal complete-core LPS provided protection against a lethal challenge with E. coli O18 LPS. The vaccine tested was nontoxic, nonpyrogenic, and immunogenic against a wide variety of pathogens found in clinical settings. PMID- 11035727 TI - Augmentation of nitric oxide production by gamma interferon in a mouse vascular endothelial cell line and its modulation by tumor necrosis factor alpha and lipopolysaccharide. AB - The effect of gamma interferon (IFN-gamma), tumor necrosis factor alpha (TNF alpha), and lipopolysaccharide (LPS) on nitric oxide (NO) production in the mouse vascular aortic endothelial cell line END-D was examined. LPS, TNF-alpha, and a low concentration of IFN-gamma inhibited NO production in END-D cells, while a high concentration of IFN-gamma definitely enhanced it. The NO production induced by a high concentration of IFN-gamma was further augmented by using IFN-gamma in combination with LPS or TNF-alpha. In sequential incubations of LPS and IFN gamma, the enhancement of NO production required prior treatment with IFN-gamma. Stimulation of END-D cells with a high concentration of IFN-gamma led to the expression of inducible NO synthase (iNOS). The augmentation of NO production by IFN-gamma alone or in combination with LPS or TNF-alpha was completely blocked by several inhibitors of iNOS. It was strongly suggested that a high concentration of IFN-gamma itself enhanced NO production in END-D cells through inducing the expression of iNOS. LPS and TNF-alpha exclusively modulated the activity of iNOS once its expression was triggered by IFN-gamma. On the other hand, a low concentration of IFN-gamma, LPS, and TNF-alpha reduced NO production through down regulating constitutive NOS (cNOS). The differential regulation of cNOS- and iNOS mediated NO production by IFN-gamma, TNF-alpha, and LPS is discussed. PMID- 11035729 TI - Lack of CD4(+) T cells does not affect induction of CD8(+) T-cell immunity against Encephalitozoon cuniculi infection. AB - Cell-mediated immunity has been reported to play an important role in defense against Encephalitozoon cuniculi infection. Previous studies from our laboratory have underlined the importance of cytotoxic CD8(+) T lymphocytes (CTL) in survival of mice infected with E. cuniculi. In the present study, immune response against E. cuniculi infection in CD4(+) T-cell-deficient mice was evaluated. Similar to resistant wild-type animals, CD4(-/-) mice were able to resolve E. cuniculi infection even at a very high challenge dose (5 x 10(7) spores/mouse). Tissues from infected CD4(-/-) mice did not exhibit higher parasite loads in comparison to the parental wild-type mice. Conversely, at day 21 postinfection, susceptible CD8(-/-) mice had 10(14) times more parasites in the liver compared to control wild-type mice. Induction of the CD8(+) T-cell response in CD4(-/-) mice against E. cuniculi infection was studied. Interestingly, a normal antigen specific CD8(+) T-cell response to E. cuniculi infection was observed in CD4(-/-) mice (precursor proliferation frequency, 1/2.5 x 10(4) versus 1/10(4) in wild type controls). Lack of CD4(+) T cells did not alter the magnitude of the antigen specific CTL response (precursor CTL frequency; 1/1.4 x 10(4) in CD4(-/-) mice versus 1/3 x 10(4) in control mice). Adoptive transfer of immune CD8(+) T cells from both CD4(-/-) and wild-type animals prevented the mortality in CD8(-/-) mice. E. cuniculi infection thus offers an example of an intracellular parasitic infection where CD8(+) T-cell immunity can be induced in the absence of CD4(+) T cells. PMID- 11035728 TI - Neisseria gonorrhoeae porin modifies the oxidative burst of human professional phagocytes. AB - A hallmark of infection with the gram-negative bacterium Neisseria gonorrhoeae is the local infiltration and subsequent activation of polymorphonuclear neutrophils. Several gonococcal outer membrane proteins are involved in the interaction with and the activation of these phagocytes, including gonococcal porin, the most abundant protein in the outer membrane. Previous work suggests that this porin plays a role in various cellular processes, including inhibiting neutrophils activation and phagosome maturation in professional phagocytes. Here we investigated the ability of porin to modify the oxidative metabolism of human peripheral blood neutrophils and monocytes in response to particulate stimuli (including live gonococci) and soluble agents. The activation of the oxidative metabolism was determined by chemiluminescence amplified with either luminol or lucigenin. We found that treatment of the phagocytes with porin inhibits the release of reactive oxygen species measured as luminol-enhanced chemiluminescence in response to zymosan, latex particles, and gonococci. The engulfment of these particles was not, however, affected by porin treatment. Similar effects of porin on the chemiluminescence response were observed in cytochalasin B-treated neutrophils exposed to the soluble chemotactic peptide N-formylmethionyl-leucyl phenylalanine. This indicates that porin selectively inhibits granule fusion with those cellular membranes that are in direct contact with porin, namely, the phagosomal and plasma membranes. This porin-induced downregulation of oxidative metabolism may be a potent mechanism by which gonococci modulate oxygen-dependent reactions by activated phagocytes at inflammation sites. PMID- 11035730 TI - Suppression of gamma interferon transcription and production by nematode excretory-secretory antigen during polyclonal stimulation of rat lymph node T cells. AB - Although certain helminth infections preferentially induce type 2 T-cell responses, the immunological mechanisms responsible for type 2 T-cell polarization remain unclear. In the present study, we investigated the effects of excretory-secretory (ES) antigen from the nematode Nippostrongylus brasiliensis on cytokine production by mesenteric lymph node (MLN) cells isolated from naive rats. MLN cells produced considerable levels of gamma interferon (IFN-gamma) during a 72-h stimulation with concanavalin A (ConA) or with immobilized anti-CD3 plus soluble anti-CD28 antibodies (anti-CD3/CD28). With either stimulation, 10 microg of ES antigen per ml significantly suppressed IFN-gamma and interleukin-2 (IL-2) production without cytotoxic activity. The copresence of anti-IL-4, anti IL-10, or transforming growth factor beta (TGF-beta) blocking antibodies did not alter the suppressive effect of ES antigen on IFN-gamma production. ES antigen did not affect IL-10 production. Kinetic studies of the effect of ES antigen indicated that the antigen suppressed even ongoing IFN-gamma production. Reverse transcription-PCR study showed that in the presence of ES antigen, IFN-gamma mRNA expression by MLN cells was suppressed 6 and 12 h after ConA or anti-CD3/CD28 stimulation. ES antigen also significantly suppressed IFN-gamma production by purified CD4(+) or CD8(+) T cells during anti-CD3/CD28 stimulation but did not affect IL-4 production by CD4(+) T cells. These findings suggested that the nematode antigen suppressed production of IFN-gamma and IL-2 but not IL-4 or IL 10 production. ES antigen-mediated suppression of IFN-gamma during the initiation of the immune response may provide a microenvironment that helps generation of type 2 T cells. PMID- 11035731 TI - Distribution of open reading frames of plasticity region of strain J99 in Helicobacter pylori strains isolated from gastric carcinoma and gastritis patients in Costa Rica. AB - The plasticity region of Helicobacter pylori strain J99 is a large chromosomal segment containing 33 strain-specific open reading frames (ORFs) with characteristics of a pathogenicity island. To study the diversity of the plasticity region, 22 probes corresponding to 20 ORFs inside the plasticity region and two ORFs on its boundaries were hybridized to genomic DNA isolated from clinical strains of H. pylori from patients with gastritis or gastric adenocarcinoma. Highly variable hybridization patterns were observed. The majority of the clinical strains presented a hybridization profile similar to that of J99; thus, these ORFs are not J99 strain specific. No association was found between a particular hybridization pattern and the clinical origin of the strain. Nevertheless, two single ORFs (JHP940 and JHP947) were more likely to be found in gastric cancer strains. They may be new pathogenicity markers. An in vitro expression study of these ORFs was also performed for the J99 strain, under different conditions. Thirteen ORFs were consistently expressed, six were consistently shut off, and three were expressed differentially. Most of the constitutionally expressed genes were located on the 3' part of the plasticity region. Our results show that the plasticity region, rather than being considered a pathogenicity island per se, should be considered a genomic island, which represents a large fragment of foreign DNA integrated into the genome and not necessarily implicated in the pathogenic capacity of the strain. PMID- 11035732 TI - Antigenic structure of outer membrane protein E of Moraxella catarrhalis and construction and characterization of mutants. AB - Outer membrane protein E (OMP E) is a 50-kDa protein of Moraxella catarrhalis which possesses several characteristics indicating that the protein will be an effective vaccine antigen. To study the antigenic structure of OMP E, eight monoclonal antibodies were developed and characterized. Three of the antibodies recognized epitopes which are present on the bacterial surface. Fusion peptides corresponding to overlapping regions of OMP E were constructed, and immunoblot assays were performed to localize the areas of the molecule bound by the monoclonal antibodies. These studies identified a surface-exposed epitope in the region of amino acids 80 through 180. To further study the protein, two mutants which lack OMP E were constructed. In bactericidal assays, the mutants were more readily killed by normal human serum compared to the isogenic parent strains. These results indicate that OMP E is involved in the expression of serum resistance of M. catarrhalis. PMID- 11035733 TI - Human neutrophil-mediated nonoxidative antifungal activity against Cryptococcus neoformans. AB - It has long been appreciated that polymorphonuclear leukocytes (PMN) kill Cryptococcus neoformans, at least in part via generation of fungicidal oxidants. The aim of this study was to examine the contribution of nonoxidative mechanisms to the inhibition and killing of C. neoformans. Treatment of human PMN with inhibitors and scavengers of respiratory burst oxidants only partially reversed anticryptococcal activity, suggesting that both oxidative and nonoxidative mechanisms were operative. To define the mediators of nonoxidative anticryptococcal activity, PMN were fractionated into cytoplasmic, primary (azurophil) granule, and secondary (specific) granule fractions. Incubation of C. neoformans with these fractions for 18 h resulted in percent inhibition of growth of 67.4 +/- 3.4, 84.6 +/- 4.4, and 29.2 +/- 10.5 (mean +/- standard error, n = 3), respectively. Anticryptococcal activity of the cytoplasmic fraction was abrogated by zinc and depletion of calprotectin. Antifungal activity of the primary granules was significantly reduced by pronase treatment, boiling, high ionic strength, and magnesium but not calcium. Fractionation of the primary granules by reverse phase high-pressure liquid chromatography on a C(4) column over an acetonitrile gradient revealed multiple peaks with anticryptococcal activity. Of these, peaks 1 and 6 had substantial fungistatic and fungicidal activity. Peak 1 was identified by acid-urea polyacrylamide gel electrophoresis (PAGE) and mass spectroscopy as human neutrophil proteins (defensins) 1 to 3. Analysis of peak 6 by sodium dodecyl sulfate-PAGE revealed multiple bands. Thus, human PMN have nonoxidative anticryptococcal activity residing principally in their cytoplasmic and primary granule fractions. Calprotectin mediates the cytoplasmic activity, whereas multiple proteins, including defensins, are responsible for activity of the primary granules. PMID- 11035734 TI - Modulation of innate cytokine responses by products of Helicobacter pylori. AB - The gastric inflammatory and immune response in Helicobacter pylori infection may be due to the effect of different H. pylori products on innate immune mechanisms. The aim of this study was to determine whether bacterial components could modulate cytokine production in vitro and thus contribute to Th1 polarization of the gastric immune response observed in vivo. The effect of H. pylori recombinant urease, bacterial lysate, intact bacteria, and bacterial DNA on proliferation and cytokine production by peripheral blood mononuclear cells (PBMCs) from H. pylori negative donors was examined as a model for innate cytokine responses. Each of the different H. pylori preparations induced gamma interferon (IFN-gamma) and interleukin-12p40 (IL-12p40), but not IL-2 or IL-5, production, and all but H. pylori DNA stimulated release of IL-10. Addition of anti-IL-12 antibody to cultures partially inhibited IFN-gamma production. In addition, each bacterial product inhibited mitogen-stimulated IL-2 production by PBMCs and Jurkat T cells. The inhibitory effect of bacterial products on IL-2 production correlated with inhibition of mitogen-stimulated lymphocyte proliferation, although urease inhibited IL-2 production without inhibiting proliferation, suggesting that inhibition of IL-2 production alone is not sufficient to inhibit lymphocyte proliferation. The results of these studies demonstrate that Th1 polarization of the gastric immune response may be due in part to the direct effects of multiple different H. pylori components that enhance IFN-gamma and IL-12 production while inhibiting both IL-2 production and cell proliferation that may be necessary for Th2 responses. PMID- 11035735 TI - Genetic dissection of primary and secondary responses to a widespread natural pathogen of the gut, Eimeria vermiformis. AB - Because most pathogens initially challenge the body at epithelial surfaces, it is important to dissect the mechanisms that underlie T-cell responses to infected epithelial cells in vivo. The coccidian parasites of the genus Eimeria are protozoan gut pathogens that elicit a potent, protective immune response in a wide range of host species. CD4+ alpha beta T cells and gamma interferon (IFN gamma) are centrally implicated in the primary immunoprotective response. To define any additional requirements for the primary response and to develop a comparison between the primary and the secondary response, we have studied Eimeria infections of a broad range of genetically altered mice. We find that a full-strength primary response depends on beta(2)-microglobulin (class I major histocompatibility complex [MHC] and class II MHC and on IFN-gamma and interleukin-6 (IL-6) but not on TAP1, perforin, IL-4, Fas ligand, or inducible nitric oxide synthetase. Indeed, MHC class II-deficient and IFN-gamma-deficient mice are as susceptible to primary infection as mice deficient in all alpha beta T cells. Strikingly, the requirements for a highly effective alpha beta-T-cell driven memory response are less stringent, requiring neither IFN-gamma nor IL-6 nor class I MHC. The class II MHC dependence was also reduced, with adoptively transferable immunity developing in MHC class II(-/-) mice. Besides the improved depiction of an immune response to a natural gut pathogen, the finding that effective memory can be elicited in the absence of primary effector responses appears to create latitude in the design of vaccine strategies. PMID- 11035736 TI - Molecular cloning and analysis of a putative siderophore ABC transporter from Staphylococcus aureus. AB - From a mass-excised Staphylococcus aureus lambdaZapII expression library, we cloned an operon encoding a novel ABC transporter with significant homology to bacterial siderophore transporter systems. The operon encodes four genes designated sstA, -B, -C, and -D encoding two putative cytoplasmic membrane proteins (sstA and sstB), an ATPase (sstC), and a membrane-bound 38-kDa lipoprotein (sstD). The sst operon is preceded by two putative Fur boxes, which indicated that expression of the sst operon was likely to be iron dependent. SstD was overexpressed in Escherichia coli, purified by Triton X-114 phase partitioning, and used to generate monospecific antisera in rats. Immunoblotting studies located SstD in the membrane fraction of S. aureus and showed that expression of the lipoprotein was reduced under iron-rich growth conditions. Triton X-114 partitioning studies on isolated membranes provided additional biochemical evidence that SstD in S. aureus is a lipoprotein. Immunoreactive polypeptides of approximately 38 kDa were detected in a wide range of staphylococcal species, but no antigenic homolog was detected in Bacillus subtilis. Expression of SstD in vivo was confirmed by immunoblotting studies with S. aureus recovered from a rat intraperitoneal chamber implant model. To further define the contribution of SstD in promoting growth of S. aureus in vitro and in vivo, we used antisense RNA technology to modulate expression of SstD. Expression of antisense sstD RNA in S. aureus resulted in a decrease in SstD expression under both iron-rich and iron-restricted growth conditions. However, this reduction in SstD levels did not affect the growth of S. aureus in vitro in an iron-limited growth medium or when grown in an intraperitoneal rat chamber implant model in vivo. PMID- 11035737 TI - Visceral leishmaniasis in mice devoid of tumor necrosis factor and response to treatment. AB - Tumor necrosis factor (TNF)-deficient mice were challenged with Leishmania donovani to characterize TNF in the response of visceral intracellular infection to antileishmanial chemotherapy. In wild-type controls (i) liver infection peaked at week 2 and resolved, (ii) discrete liver granulomas developed at weeks 2 to 4 and involuted, and (iii) leishmanicidal responses to antimony (Sb), amphotericin B (AmB), and miltefosine were intact. In TNF knockout (KO) mice (i) initial liver infection was unrestrained, plateaued, and then declined somewhat by week 6, (ii) an absent early granulomatous reaction abruptly accelerated with striking tissue inflammation, widespread hepatic necrosis, and 100% mortality by week 10, and (iii) while the initial response to AmB and miltefosine was intact, killing induced by Sb therapy was reduced by >50%. Although initial AmB treatment during weeks 2 to 3 killed 98% of liver parasites, 75% of AmB-treated KO mice subsequently relapsed and died by week 12; however, additional maintenance AmB preserved long-term survival. These results for a model of visceral infection indicate that endogenous TNF is required early on to control intracellular L. donovani, support granuloma development, and mediate optimal initial effects of Sb and prevent relapse after ordinarily curative AmB treatment. A compensatory, TNF-independent antileishmanial mechanism developed in TNF KO mice; however, its effect was uncontrolled fatal inflammation. Chemotherapeutic elimination of the parasite stimulus reversed the hyperinflammatory response and preserved survival. PMID- 11035738 TI - Mononuclear cell recruitment, granuloma assembly, and response to treatment in experimental visceral leishmaniasis: intracellular adhesion molecule 1-dependent and -independent regulation. AB - In experimental visceral leishmaniasis, acquired resistance to intracellular Leishmania donovani is Th1 cell cytokine dependent and largely mediated by gamma interferon (IFN-gamma); the same response also permits conventional antimony (Sb) chemotherapy to express its leishmanicidal effect. Since the influxing blood monocyte (which utilizes endothelial cell ICAM-1 for adhesion and tissue entry) is a primary effector target cell for this cytokine mechanism, we tested the monocyte's role in host responsiveness to chemotherapy in mice with ICAM-1 gene disruptions. Mutant animals failed to develop any early granulomatous tissue response in the liver, initially supported high-level visceral parasite replication, and showed no killing after Sb treatment; the leishmanicidal response to a directly acting, alternative chemotherapeutic probe, amphotericin B, was intact. However, mutant mice proceeded to express a compensatory, ICAM-1 independent response leading to mononuclear cell influx and granuloma assembly, control over visceral infection, and the capacity to respond to Sb. Together, these results point to the recruitment of emigrant monocytes and mononuclear cell granuloma formation, mediated by ICAM-1-dependent and -independent pathways, as critical determinants of host responsiveness to conventional antileishmanial chemotherapy. PMID- 11035739 TI - Necrosis of lung epithelial cells during infection with Mycobacterium tuberculosis is preceded by cell permeation. AB - Mycobacterium tuberculosis establishes infection, progresses towards disease, and is transmitted from the alveolus of the lung. However, the role of the alveolar epithelium in any of these pathogenic processes of tuberculosis is unclear. In this study, lung epithelial cells (A549) were used as a model in which to examine cytotoxicity during infection with either virulent or avirulent mycobacteria in order to further establish the role of the lung epithelium during tuberculosis. Infection of A549 cells with M. tuberculosis strains Erdman and CDC1551 demonstrated significant cell monolayer clearing, whereas infection with either Mycobacterium bovis BCG or Mycobacterium smegmatis LR222 did not. Clearing of M. tuberculosis-infected A549 cells correlated to necrosis, not apoptosis. Treatment of M. tuberculosis-infected A549 cells with streptomycin, but not cycloheximide, demonstrated a significant reduction in the necrosis of A549 cell monolayers. This mycobacterium-induced A549 necrosis did not correlate to higher levels of intracellular or extracellular growth by the mycobacteria during infection. Staining of infected cells with propidium iodide demonstrated that M. tuberculosis induced increased permeation of A549 cell membranes within 24 h postinfection. Quantitation of lactate dehydrogenase (LDH) release from infected cells further demonstrated that cell permeation was specific to M. tuberculosis infection and correlated to A549 cellular necrosis. Inactivated M. tuberculosis or its subcellular fractions did not result in A549 necrosis or LDH release. These studies demonstrate that lung epithelial cell cytotoxicity is specific to infection by virulent mycobacteria and is caused by cellular necrosis. This necrosis is not a direct correlate of mycobacterial growth or of the expression of host cell factors, but is preceded by permeation of the A549 cell membrane and requires infection with live bacilli. PMID- 11035740 TI - Salmonella enterica serovar typhimurium-induced maturation of bone marrow-derived dendritic cells. AB - Murine bone marrow-derived dendritic cells (DC) can phagocytose and process Salmonella enterica serovar Typhimurium for peptide presentation on major histocompatibility complex class I (MHC-I) and MHC-II molecules. To investigate if a serovar Typhimurium encounter with DC induces maturation and downregulates their ability to present antigens from subsequently encountered bacteria, DC were pulsed with serovar Typhimurium 24 h prior to coincubating with Escherichia coli expressing the model antigen Crl-OVA. Quantitating presentation of OVA epitopes contained within Crl-OVA showed that Salmonella-pulsed DC had a reduced capacity to process Crl-OVA-expressing E. coli for OVA(257-264)/K(b) and OVA(265-277)/I A(b) presentation. In addition, time course studies of DC pulsed with Crl-OVA expressing serovar Typhimurium showed that OVA(257-264)/K(b) complexes could stimulate CD8OVA T-hybridoma cells for <24 h following a bacterial pulse, while OVA(265-277)/I-A(b) complexes could stimulate OT4H T-hybridoma cells for >24 but <48 h. The phoP-phoQ virulence locus of serovar Typhimurium also influenced the ability of DC to process Crl-OVA-expressing serovar Typhimurium for OVA(265 277)/I-A(b) presentation but not for OVA(257-264)/K(b) presentation. Furthermore, pulsing of DC with serovar Typhimurium followed by incubation for 24 or 48 h altered surface expression of MHC-I, MHC-II, CD40, CD54, CD80, and CD86, generating a DC population with a uniform, high expression level of these molecules. Finally, neither the serovar Typhimurium phoP-phoQ locus nor lipopolysaccharides (LPS) containing lipid A modifications purified from phoP mutant strains had a different effect on DC maturation from that of wild-type serovar Typhimurium or purified wild-type LPS. Thus, these data show that Salmonella or Salmonella LPS induces maturation of DC and that this process is not altered by the Salmonella phoP virulence locus. However, phoP did influence OVA(265-277)/I-A(b) presentation by DC infected with Crl-OVA-expressing serovar Typhimurium when quantitated after 2 h of bacterial infection. PMID- 11035741 TI - Staphylococcal fibronectin binding protein interacts with heat shock protein 60 and integrins: role in internalization by epithelial cells. AB - We reported previously that internalization of Staphylococcus aureus by nonprofessional phagocytes involves an interaction between fibronectin (Fn) binding protein (FnBP) and the host cell, resulting in signal transduction, tyrosine kinase activity, and cytoskeletal rearrangement (K. Dziewanowska, J. M. Patti, C. F. Deobald, K. W. Bayles, W. R. Trumble, and G. A. Bohach, Infect. Immun. 67:4673-4678, 1999). The goal of the present study was to identify the host molecules responsible for uptake of the organism through an interaction with FnBP. First, Fn was required for internalization. Addition of small amounts of exogenous Fn stimulated the uptake of S. aureus by HEp-2 cells, which are deficient in Fn synthesis. Fn antibodies blocked internalization of the organism by MAC-T cell monolayers, a bovine epithelial cell line which expresses Fn. Second, a monoclonal antibody (MAb) specific for beta(1) integrins dramatically reduced S. aureus invasion, suggesting that the formation of a Fn bridge linking the host cell beta(1) integrin and FnBP precedes internalization. However, ligand blotting of cell membrane proteins with a functional fragment of FnBP consistently identified an additional approximately 55-kDa receptor on both human and bovine epithelial cells. This protein was purified and identified by N terminal microsequencing as heat shock protein 60 (Hsp60). The interaction between FnBP and Hsp60 also occurred when the whole cells were used. Cell membrane localization of Hsp60 was confirmed by biotinylation with an agent nonpermeable to the cell membrane. Pretreatment of epithelial cells with a MAb specific for eukaryotic Hsp60 significantly reduced internalization of S. aureus. Combined, these results suggest that the FnBP binds directly to both Hsp60 and Fn and is linked to beta(1) integrins through a Fn bridge. The simultaneous involvement of Fn and two host cell ligands, beta(1) integrins and Hsp60, suggests that FnBP is a multifunctional adhesin that mediates internalization in a manner similar to that proposed for OpaA, the Neisseria gonorrhoeae FnBP homolog (J. P. M. van Putten, T. D. Duensing, and R. L. Cole, Mol. Microbiol. 29:369-379, 1998). PMID- 11035742 TI - TonB is required for intracellular growth and virulence of Shigella dysenteriae. AB - To assess the importance of TonB-dependent iron transport systems to growth of Shigella in vivo, a tonB mutant of Shigella dysenteriae was isolated and tested in cultured cells. The tonB mutant invaded epithelial cells, but did not form plaques in confluent monolayers of Henle cells, indicating an inability of this mutant to spread from cell to cell. The rate of intracellular multiplication of the tonB mutant was reduced significantly compared to that of the wild type. The loss of virulence in the tonB mutant was not due to loss of either Shu or Ent, the TonB-dependent systems which allow for transport of heme and ferrienterobactin, respectively. A shuA mutant lacking the outer membrane receptor for heme, an entB mutant defective in enterobactin synthesis, and a shuA entB double mutant each were able to invade cultured cells, multiply intracellularly, and form wild-type plaques. The ability of S. dysenteriae to access iron during intracellular growth was assessed by flow cytometric analysis of an iron- and Fur-regulated shuA-gfp reporter construct. Low levels of green fluorescent protein expression in the intracellular environment were observed in all strains, indicating that iron is available to intracellular bacteria, even in the absence of TonB-dependent iron transport. The failure of the tonB mutant to grow well in an iron-replete intracellular environment suggests that TonB plays a role in addition to heme- and siderophore-mediated iron acquisition in vivo, and this function is required for the intracellular growth and intercellular spread of S. dysenteriae. PMID- 11035743 TI - Role of catalase in Campylobacter jejuni intracellular survival. AB - The ability of Campylobacter jejuni to penetrate normally nonphagocytic host cells is believed to be a key virulence determinant. Recently, kinetics of C. jejuni intracellular survival have been described and indicate that the bacterium can persist and multiply within epithelial cells and macrophages in vitro. Studies conducted by Pesci et al. indicate that superoxide dismutase contributes to intraepithelial cell survival, as isogenic sod mutants are 12-fold more sensitive to intracellular killing than wild-type strains. These findings suggest that bacterial factors that combat reactive oxygen species enable the organism to persist inside host cells. Experiments were conducted to determine the contribution of catalase to C. jejuni intracellular survival. Zymographic analysis indicated that C. jejuni expresses a single catalase enzyme. The gene encoding catalase (katA) was cloned via functional complementation, and an isogenic katA mutant strain was constructed. Kinetic studies indicate that catalase provides resistance to hydrogen peroxide in vitro but does not play a role in intraepithelial cell survival. Catalase does however contribute to intramacrophage survival. Kinetic studies of C. jejuni growth in murine and porcine peritoneal macrophages demonstrated extensive killing of both wild-type and katA mutant strains shortly following internalization. Long-term cultures (72 h postinfection) of infected phagocytes permitted recovery of viable wild-type C. jejuni; in contrast, no viable katA mutant bacteria were recovered. Accordingly, inhibition of macrophage nitric oxide synthase or NADPH oxidase permitted recovery of katA mutant C. jejuni. These observations indicate that catalase is essential for C. jejuni intramacrophage persistence and growth and suggest a novel mechanism of intracellular survival. PMID- 11035744 TI - Identification of polymorphonuclear leukocyte and HL-60 cell receptors for adhesins of Streptococcus gordonii and Actinomyces naeslundii. AB - Interactions of oral streptococci and actinomyces with polymorphonuclear leukocytes (PMNs), mediated by sialic acid- and Gal/GalNAc-reactive adhesins, respectively, result in activation of the PMNs and thereby may contribute to the initiation of oral inflammation. Sialidase treatment of PMNs or HL-60 cells abolished adhesion of Streptococcus gordonii but was required for adhesion of Actinomyces naeslundii. The same effects of sialidase were noted for adhesion of these bacteria to a major 150-kDa surface glycoprotein of either PMNs or undifferentiated HL-60 cells and to a 130-kDa surface glycoprotein of differentiated HL-60 cells. These glycoproteins were both identified as leukosialin (CD43) by immunoprecipitation with a specific monoclonal antibody (MAb). Adhesion of streptococci and actinomyces to a 200-kDa minor PMN surface glycoprotein was also detected by bacterial overlay of untreated and sialidase treated nitrocellulose transfers, respectively. This glycoprotein was identified as leukocyte common antigen (CD45) by immunoprecipitation with a specific MAb. CD43 and CD45 both possess extracellular mucinlike domains in addition to intracellular domains that are implicated in signal transduction. Consequently, the interactions of streptococci and actinomyces with the mucinlike domains of these mammalian cell surface glycoproteins result not only in adhesion but, in addition, may represent the initial step in PMN activation by these bacteria. PMID- 11035745 TI - Essential role of platelet-activating factor in control of Leishmania (Leishmania) amazonensis infection. AB - In the present study we investigated the role of platelet-activating factor (PAF) and prostaglandins in experimental Leishmania (Leishmania) amazonensis infection and the relationship between these mediators and nitric oxide (NO) production. Mouse peritoneal macrophages elicited with thioglicolate were infected with leishmania amastigotes, and the infection index determined 48 h later. The course of infection was monitored for 5 weeks in mice infected in the footpad with promastigotes by measuring the footpad swelling and parasite load in regional lymph nodes and spleen. The addition of PAF to C57BL/6 mouse macrophages significantly inhibited parasite growth and induced NO production. Treatment of macrophages with a selective PAF antagonist, WEB2086, increased the infection, indicating that endogenously produced PAF regulates macrophage ability to control leishmania infection. This effect of PAF was abolished by addition of the inhibitor of NO synthesis, L-NAME, to the cultures. The addition of prostaglandin E(2) significantly increased the infection and NO production. Treatment with cyclo-oxygenase inhibitor, indomethacin, reduced the infection and PAF-induced release of NO. Thus, the increased NO production induced by PAF seems to be mediated by prostaglandins. The more-selective inhibitors of cyclo-oxygenase 2, nimesulide and NS-398, had no significant effect. Thus, antileishmanial activity correlates better with the presence of PAF or absence of prostaglandins than with NO production. In vivo treatment with PAF antagonists significantly increased leishmania lesions, as well as the parasite load, in regional lymph nodes and spleens. These findings indicate that PAF is essential for the control of leishmania infection. PMID- 11035746 TI - Inverse relation between disease severity and expression of the streptococcal cysteine protease, SpeB, among clonal M1T1 isolates recovered from invasive group A streptococcal infection cases. AB - The streptococcal cysteine protease (SpeB) is one of the major virulence factors produced by group A streptococci (GAS). In this study we investigated if differences exist in SpeB production by clonally related M1T1 clinical isolates derived from patients with invasive infections. Twenty-nine of these isolates were from nonsevere cases and 48 were from severe cases, including streptococcal toxic shock syndrome (STSS) and necrotizing fasciitis (NF) cases. The expression and amount of the 28-kDa SpeB protein produced were determined by quantitative Western blotting, and protease activity was measured by a fluorescent enzymatic assay. A high degree of variation in SpeB expression was seen among the isolates, and this variation seemed to correlate with the severity and/or clinical manifestation of the invasive infection. The mean amount of 28-kDa SpeB protein and cysteine protease activity produced by isolates from nonsevere cases was significantly higher than that from STSS cases (P = 0.001). This difference was partly due to the fact that 41% of STSS isolates produced little or no SpeB compared to only 14% of isolates recovered in nonsevere cases. Moreover, the cysteine protease activity among those isolates that expressed SpeB was significantly lower for STSS isolates than for isolates from nonsevere cases (P = 0.001). Increased SpeB production was also inversely correlated with intact M protein expression, and inhibition of cysteine protease activity blocked the cleavage of the surface M protein. Together, the data support the existence of both an "on-off" and a posttranslational regulatory mechanism(s) controlling SpeB production, and they suggest that isolates with the speB gene in the "off" state are more likely to spare the surface M protein and to be isolated from cases of severe rather than nonsevere invasive infection. These findings may have important implications for the role of SpeB in host-pathogen interactions via regulation of the expression of GAS virulence genes and the severity of invasive disease. PMID- 11035747 TI - SclA, a novel collagen-like surface protein of Streptococcus pyogenes. AB - Surface proteins of Streptococcus pyogenes are important virulence factors. Here we describe a novel collagen-like surface protein, designated SclA (streptococcal collagen-like surface protein). The sclA gene was identified in silico using the Streptococcal Genome Sequencing Project with the recently identified protein GRAB as the probe. SclA has a signal sequence and a cell wall attachment region containing the prototypic LPXTGX motif. The surface-exposed part of SclA contains a unique NH(2)-terminal domain of 73 amino acids, followed by a collagen-like region. The sclA gene was found to be positively regulated by Mga, a transcriptional activator of several S. pyogenes virulence determinants. A mutant lacking cell wall-associated SclA was constructed and was found to be as effective as wild-type bacteria in platelet aggregation, survival in fresh human blood, and adherence to pharyngeal cells. The sclA gene was found in all 12 S. pyogenes strains that were investigated using PCR. Sequence analysis revealed that the signal sequence and the cell wall attachment region are highly conserved. The collagen-like domain is variable in its NH(2)-terminal region and has conserved repeated domains in its COOH-terminal part. SclA proteins from most strains have additional proline-rich repeats spacing the collagen-like domain and the cell wall attachment sequence. The unique NH(2)-terminal region is hypervariable, but computer predictions indicate a common secondary structure, with two alpha helices connected by a loop region. Immune selection may explain the hypervariability in the NH(2)-terminal region, whereas the preserved secondary structure implies that this region has a common function. These features and the Mga regulation are shared with the M protein of S. pyogenes. Moreover, as with the gene encoding the M protein, phylogenetic analysis indicates that horizontal gene transfer has contributed to the evolution of sclA. PMID- 11035748 TI - Characterization of the catalytic domain of Clostridium novyi alpha-toxin. AB - Clostridium novyi alpha-toxin belongs to the family of large clostridial cytotoxins which act on cells through the modification of small GTP-binding proteins. We present here an analysis of the catalytic domain of alpha-toxin. A NH(2)-terminal 551-amino-acid fragment, alpha 551, was found to contain the full enzyme activity of the holotoxin, whereas a slightly shortened fragment encompassing 509 amino acids showed no detectable enzyme activity. Further characterization of the enzymatically active fragment alpha 551 revealed a substrate specificity for both UDP-N-acetylglucosamine and UDP-glucose. A Michaelis-Menten constant of 17 microM was determined for the substrate UDP-N acetylglucosamine, while that for UDP-glucose was about 20 times higher, indicating a weaker affinity of the toxin for the latter substrate. Mutation of the aspartic acids of a conserved motif DXD within alpha 551 reduced enzyme activity >700-fold and inhibited cytotoxicity after microinjection in cells. Inhibition of enzyme activity of the DXD mutant could be partially overcome by increased concentrations of manganese ions, suggesting the involvement of these aspartic acids in Mn(2+) binding. By construction of chimeras of enzymatically active fragments of C. sordellii lethal toxin and C. novyi alpha-toxin, we located the region involved in nucleotide-sugar specificity to amino acids 133 through 517. PMID- 11035749 TI - Role of streptolysin O in a mouse model of invasive group A streptococcal disease. AB - Many of the virulence factors that have been characterized for group A streptococci (GAS) are not expressed in all clinical isolates. One putative virulence factor that is present among most is streptolysin O (Slo), a protein with well-characterized cytolytic activity for many eukaryotic cells types. In other bacterial pathogens, proteins homologous to Slo have been shown to be essential for virulence, but the role of Slo in GAS had not been previously examined. To investigate the role of Slo in GAS virulence, we examined both revertible and stable slo mutants in a mouse model of invasive disease. When the revertible slo mutant was used to infect mice, the reversion frequency of bacteria isolated from the wounds and spleens of infected animals was more than 100 times that of the inoculum, indicating that there was selective pressure in the animal for Slo(+) GAS. Experiments with the stable slo mutant demonstrated that Slo was not necessary for the formation of necrotic lesions, nor was it necessary for escape from the lesion to cause disseminated infection. Bacteria were present in the spleens of 50% of the mice that survived infection with the stable slo mutant, indicating that dissemination of Slo(-) GAS does not always cause disease. Finally, mice infected with the stable slo mutant exhibited a significant decrease in mortality rates compared to mice infected with wild-type GAS (P < 0.05), indicating that Slo plays an important role in GAS virulence. PMID- 11035750 TI - Construction of a Vibrio cholerae vaccine candidate using transposon delivery and FLP recombinase-mediated excision. AB - Recent efforts to develop a vaccine against the diarrheal disease cholera have focused on the use of live attenuated strains of the causative organism, Vibrio cholerae. The Ogawa lipopolysaccharide phenotype is expressed by many epidemic strains, and motility defects reduce the risk of reactive diarrhea in vaccine recipients. We therefore converted a motile Inaba(+) vaccine candidate, Peru-2, to a nonmotile Ogawa(+) phenotype using a mariner-based transposon carrying rfbT, the gene required for expression of the Ogawa phenotype. Analysis of 22 nonmotile Peru-2 mutants showed that two were Ogawa(+), and both of these strains had insertions in the flgE gene. It was possible to convert these strains to antibiotic sensitivity by introducing a recombinase that acts on sites flanking the antibiotic marker on the transposon. The resulting strains are competent for colonization in infant mice and may therefore be suitable as vaccine candidates for use either independently or in a combination with strains of different biotypes and serotypes. PMID- 11035751 TI - A novel polymorphism in the toll-like receptor 2 gene and its potential association with staphylococcal infection. AB - The toll-like receptor 2 (TLR2) has gained importance as a major mammalian receptor for lipoproteins derived from the cell wall of a variety of bacteria, such as Borrelia burgdorferi, Treponema pallidum, and Mycoplasma fermentans. We were interested in identifying mutations in the TLR2 gene that might prove to be associated with altered susceptibility to septic shock. We performed a mutation screen of the TLR2 gene using single-stranded conformational polymorphism in 110 normal, healthy study subjects and detected an Arg753Gln mutation in three individuals. No other missense mutations were detected in the TLR2 open reading frame. Functional studies demonstrate that the Arg753Gln polymorphism, in comparison to the wild-type TLR2 gene, is significantly less responsive to bacterial peptides derived from B. burgdorferi and T. pallidum. In a septic shock population, the Arg753Gln TLR2 polymorphism occurred in 2 out of 91 septic patients. More importantly, both of the subjects with the TLR2 Arg753Gln polymorphism had staphylococcal infections. These findings suggest that a mutation in the TLR2 gene may predispose individuals to life-threatening bacterial infections. PMID- 11035753 TI - Construction and characterization of a nonproliferative El Tor cholera vaccine candidate derived from strain 638. AB - In recent clinical assays, our cholera vaccine candidate strain, Vibrio cholerae 638 El Tor Ogawa, was well tolerated and immunogenic in Cuban volunteers. In this work we describe the construction of 638T, a thymidine auxotrophic version of improved environmental biosafety. In so doing, the thyA gene from V. cholerae was cloned, sequenced, mutated in vitro, and used to replace the wild-type allele. Except for its dependence on thymidine for growth in minimal medium, 638T is essentially indistinguishable from 638 in the rate of growth and morphology in complete medium. The two strains showed equivalent phenotypes with regard to motility, expression of the celA marker, colonization capacity in the infant mouse cholera model, and immunogenicity in the adult rabbit cholera model. However, the ability of this new strain to survive environmental starvation was limited with respect to that of 638. Taken together, these results suggest that this live, attenuated, but nonproliferative strain is a new, promising cholera vaccine candidate. PMID- 11035752 TI - Identification of tgh-2, a filarial nematode homolog of Caenorhabditis elegans daf-7 and human transforming growth factor beta, expressed in microfilarial and adult stages of Brugia malayi. AB - A novel member of the transforming growth factor beta (TGF-beta) family has been identified in the filarial nematode parasite Brugia malayi by searching the recently developed Expressed Sequence Tag (EST) database produced by the Filarial Genome Project. Designated tgh-2, this new gene shows most similarity to a key product regulating dauer larva formation in Caenorhabditis elegans (DAF-7) and to the human down-modulatory cytokine TGF-beta. Homology to DAF-7 extends throughout the length of the 349-amino-acid (aa) protein, which is divided into an N terminal 237 aa, including a putative signal sequence, a 4-aa basic cleavage site, and a 108-aa C-terminal active domain. Similarity to human TGF-beta is restricted to the C-terminal domain, over which there is a 32% identity between TGH-2 and TGF-beta1, including every cysteine residue. Expression of tgh-2 mRNA has been measured over the filarial life cycle. It is maximal in the microfilarial stage, with lower levels of activity around the time of molting within the mammal, but continues to be expressed by mature adult male and female parasites. Expression in both the microfilaria, which is in a state of arrested development, and the adult, which is terminally differentiated, indicates that tgh-2 may play a role other than purely developmental. This is consistent with our observation that TGH-2 is secreted by adult worms in vitro. Recombinant TGH-2 expressed in baculovirus shows a low level of binding to TGF-beta-receptor bearing mink lung epithelial cells (MELCs), which is partially inhibited (16 to 39%) with human TGF-beta, and activates plasminogen activator inhibitor-1 transcription in MELCs, a marker for TGF-beta-mediated transduction. Further tests will be required to establish whether the major role of B. malayi TGH-2 (Bm TGH-2) is to modulate the host immune response via the TGF-beta pathway. PMID- 11035754 TI - Hemolytic and hemoxidative activities in Mycoplasma penetrans. AB - Mycoplasma penetrans is a newly isolated Mollicute from the urine of patients infected with human immunodeficiency virus that demonstrates the capacity to adhere to and invade human cells. A previous report, based on assays with mouse red blood cells (RBCs), indicated that M. penetrans lacked hemolytic activity. In our studies, we incubated different isolates of M. penetrans with various RBC species and observed hemolytic zones surrounding individual mycoplasma colonies. All M. penetrans strains displayed hemolysis after 2 to 3 days of incubation. Hemolytic activity diffused from single colonies, eventually causing complete lysis. Hemolysis was most pronounced with sheep RBCs, followed by horse, chicken, and human cells. Furthermore, hemolytic activity was demonstrable in both intact mycoplasma cell preparations and spent culture supernatant. However, unlike intact mycoplasmas, the hemolytic activity in the supernatant was dependent on the reducing agent, cysteine. In addition to hemolysis, a brown precipitate was closely associated with mycoplasma colonies, suggesting oxidation of hemoglobin. Absorption spectra indicated that hemoglobin was oxidized to methemoglobin, and the addition of catalase demonstrated H(2)O(2)-mediated hemoxidation. Other experiments suggested that hemoxidation enhanced total hemolysis, providing the first evidence of both hemolytic and hemoxidative activities in M. penetrans. PMID- 11035755 TI - Involvement of focal adhesion kinase in Escherichia coli invasion of human brain microvascular endothelial cells. AB - Escherichia coli K1 traversal across the blood-brain barrier is an essential step in the pathogenesis of neonatal meningitis. We have previously shown that invasive E. coli promotes the actin rearrangement of brain microvascular endothelial cells (BMEC), which constitute a lining of the blood-brain barrier, for invasion. However, signal transduction mechanisms involved in E. coli invasion are not defined. In this report we show that tyrosine kinases play a major role in E. coli invasion of human BMEC (HBMEC). E. coli induced tyrosine phosphorylation of HBMEC cytoskeletal proteins, focal adhesion kinase (FAK), and paxillin, with a concomitant increase in the association of paxillin with FAK. Overexpression of a dominant interfering form of the FAK C-terminal domain, FRNK (FAK-related nonkinase), significantly inhibited E. coli invasion of HBMEC. Furthermore, we found that FAK kinase activity and the autophosphorylation site (Tyr397) are important in E. coli invasion of HBMEC, whereas the Grb2 binding site (Tyr925) is not required. Immunocytochemical studies demonstrated that FAK is recruited to focal plaques at the site of bacterial entry. Consistent with the invasion results, overexpression of FRNK, a kinase-negative mutant (Arg454 FAK), and a Src binding mutant (Phe397 FAK) inhibited the accumulation of FAK at the bacterial entry site. The overexpression of FAK mutants in HBMEC also blocked the E. coli-induced tyrosine phosphorylation of FAK and its association with paxillin. These observations provide evidence that FAK tyrosine phosphorylation and its recruitment to the cytoskeleton play a key role in E. coli invasion of HBMEC. PMID- 11035756 TI - Temporal pore formation-mediated egress from macrophages and alveolar epithelial cells by Legionella pneumophila. AB - Legionella pneumophila does not induce apoptosis in the protozoan host, but induces pore formation-mediated cytolysis after termination of intracellular replication (L.-Y. Gao and Y. Abu Kwaik, Environ. Microbiol. 2:79-90, 2000). In contrast to this single mode of killing of protozoa, we have recently proposed a biphasic model by which L. pneumophila kills macrophages, in which the first phase is manifested through the induction of apoptosis during early stages of the infection, followed by an independent and temporal induction of necrosis during late stages of intracellular replication. Here we show that, similar to the protozoan host, the induction of necrosis and cytolysis of macrophages by L. pneumophila is mediated by the pore-forming toxin or activity. This activity is temporally and maximally expressed only upon termination of bacterial replication and correlates with cytolysis of macrophages and alveolar epithelial cells in vitro. We have identified five L. pneumophila mutants defective in the pore forming activity. The phagosomes harboring the mutants do not colocalize with the late endosomal or lysosomal marker Lamp-1, and the mutants replicate intracellularly similar to the parental strain. Interestingly, despite their prolific intracellular replication, the mutants are defective in cytotoxicity and are "trapped" within and fail to lyse and egress from macrophages and alveolar epithelial cells upon termination of intracellular replication. However, the mutants are subsequently released from the host cell, most likely due to apoptotic death of the host cell. Data derived from cytotoxicity assays, confocal laser scanning microscopy, and electron microscopy confirm the defect in the mutants to induce necrosis of macrophages and the failure to egress from the host cell. Importantly, the mutants are completely defective in acute lethality (24 to 48 h) to intratracheally inoculated A/J mice. We conclude that the pore-forming activity of L. pneumophila is not required for phagosomal trafficking or for intracellular replication. This activity is expressed upon termination of bacterial replication and is essential to induce cytolysis of infected macrophages to allow egress of intracellular bacteria. In addition, this activity plays a major role in pulmonary immunopathology in vivo. PMID- 11035757 TI - Expression of peptidoglycan-associated lipoprotein is required for virulence in the human model of Haemophilus ducreyi infection. AB - Haemophilus ducreyi expresses a peptidoglycan-associated lipoprotein (PAL) that exhibits extensive homology to Haemophilus influenzae protein 6. We constructed an isogenic PAL mutant (35000HP-SMS4) by the use of a suicide vector that contains lacZ as a counterselectable marker. H. ducreyi 35000HP-SMS4 and its parent, 35000HP, had similar growth rates in broth and similar lipooligosaccharide profiles. 35000HP-SMS4 formed smaller, more transparent colonies than 35000HP and, unlike its parent, was hypersensitive to antibiotics. Complementation of the mutant in trans restored the parental phenotypes. To test whether expression of PAL is required for virulence, nine human volunteers were experimentally infected. Each subject was inoculated with two doses (41 to 89 CFU) of live 35000HP and one dose of heat-killed bacteria on one arm and with three doses (ranging from 28 to 800 CFU) of live 35000HP-SMS4 on the other arm. Papules developed at similar rates at sites inoculated with the mutant or parent but were significantly smaller at mutant-inoculated sites than at parent inoculated sites. The pustule formation rate was 72% (95% confidence interval [CI], 46.5 to 90.3%) at 18 parent sites and 11% (95% CI, 2.4 to 29.2%) at 27 mutant sites (P < 0.0001). The rates of recovery of H. ducreyi from surface cultures were 8% (n = 130; 95% CI, 4.3 to 14.6%) for parent-inoculated sites and 0% (n = 120; 95% CI, 0.0 to 2.5%) for mutant-inoculated sites (P < 0.001). H. ducreyi was recovered from six of seven biopsied parent-inoculated sites and from one of three biopsied mutant-inoculated sites. Confocal microscopy confirmed that the bacteria present in a mutant inoculation site pustule lacked a PAL-specific epitope. Although biosafety regulations precluded our testing the complemented mutant in humans, these results suggest that expression of PAL facilitates the ability of H. ducreyi to progress to the pustular stage of disease. PMID- 11035758 TI - Key role for DsbA in cell-to-cell spread of Shigella flexneri, permitting secretion of Ipa proteins into interepithelial protrusions. AB - DsbA, a disulfide bond catalyst, is necessary for realization of the pathogenic potential of Shigella flexneri. Sh42, a mutant strain differing from wild-type M90TS solely because it expresses nonfunctional DsbA33G (substitution for 33C at the active site), secreted less IpaB and IpaC than M90TS in response to various stimuli in vitro. A kinetic study demonstrated that Sh42 responded more slowly to Congo red than M90TS. By modulating relative concentrations of functional and nonfunctional DsbA within bacteria, functional enzyme has been shown to be necessary for intercellular spread. By confocal microscopy, M90TS dividing in protrusions was shown to secrete Ipa proteins from the septation furrow, anticipating lysis of protrusions, while Sh42 showed minimal Ipa secretion in this location. In the light of a previous demonstration that DsbA is not necessary for entry of epithelial cells, we conclude that a role in virulence of this disulfide bond catalyst lies in facilitating secretion of Ipa proteins specifically within epithelial protrusions, in turn allowing cell-to-cell spread of S. flexneri. PMID- 11035759 TI - Evidence for vaccine synergy between Borrelia burgdorferi decorin binding protein A and outer surface protein A in the mouse model of lyme borreliosis. AB - Mice immunized with either the predominantly vector-stage lipoprotein outer surface protein A (OspA) or the in vivo-expressed lipoprotein decorin binding protein A (DbpA) are protected against Borrelia burgdorferi challenge. DbpA-OspA combinations protected against 100-fold-higher challenge doses than did either single-antigen vaccine and conferred significant protection against heterologous B. burgdorferi, B. garinii, and B. afzelii isolates, suggesting that there is synergy between these two immunogens. PMID- 11035760 TI - Micronemal transport of Plasmodium ookinete chitinases to the electron-dense area of the apical complex for extracellular secretion. AB - Plasmodium ookinetes secrete chitinases to penetrate the acellular, chitin containing peritrophic matrix of the mosquito midgut en route to invasion of the epithelium. Chitinases are potentially targets that can be used to block malaria transmission. We demonstrate here that chitinases of Plasmodium falciparum and P. gallinaceum are concentrated at the apical end of ookinetes. The chitinase PgCHT1 of P. gallinaceum is present within ookinete micronemes and subsequently becomes localized in the electron-dense area of the apical complex. These observations suggest a pathway by which ookinetes secrete proteins extracellularly. PMID- 11035761 TI - Yersinia pestis YscG protein is a Syc-like chaperone that directly binds yscE. AB - Pathogenic Yersinia species secrete virulence proteins, termed Yersinia outer proteins (Yops), upon contact with a eukaryotic cell. The secretion machinery is composed of 21 Yersinia secretion (Ysc) proteins. Yersinia pestis mutants defective in expression of YscG or YscE were unable to export the Yops. YscG showed structural and limited amino-acid-sequence similarities to members of the specific Yop chaperone (Syc) family of proteins. YscG specifically recognized and bound YscE; however, unlike previously characterized Syc substrates, YscE was not exported from the cell. These data suggest that YscG functions as a chaperone for YscE. PMID- 11035762 TI - Contribution of plasmid-encoded fimbriae and intimin to capacity of rabbit specific enteropathogenic Escherichia coli to attach to and colonize rabbit intestine. AB - Attachment to the intestinal mucosa is an essential step in the pathogenesis of diarrhea caused by enteropathogenic Escherichia coli (EPEC). Fimbriae and intimin, the outer membrane protein product of the chromosomal eae gene, contribute to this process, but their relative roles and the nature of their interaction are not known. The aim of this study was to determine the relative contribution of plasmid-encoded fimbriae, termed Ral, and intimin to the capacity of rabbit-specific EPEC (REPEC) to attach to the intestinal mucosa of rabbits. To achieve this, we constructed a series of mutants in REPEC strain 83/39 (O15:H-), in which the ralE and eae genes were insertionally inactivated. These strains were then inoculated into ligated loops of rabbit ileum, which were resected 18 h later and examined by light and electron microscopy. The results showed that intimin, but not Ral, is essential for the elicitation of attaching-effacing lesions by REPEC. Nevertheless, a delta eae Ral-bearing mutant adhered to the intestinal epithelium to the same extent as its eae-positive parent and far more extensively than an eae(+) delta ral strain. To examine the contribution of Ral and intimin to colonization of rabbit intestine, we fed these strains to weanling rabbits, which were killed 4 days later, so that the number of bacteria in various regions of the intestine could be determined. The results indicated that strain 83/39 requires both Ral and intimin to colonize the intestine successfully and that a delta eae delta ralE double mutant was incapable of colonizing the intestine. Taken together, these findings indicate that Ral and intimin act independently as adhesion factors of REPEC strain 83/39 and that this strain carries no other significant colonization factor. When both Ral and intimin are present, they appear to act cooperatively, with Ral-mediated adhesion preceding that mediated by intimin. PMID- 11035763 TI - Inhibition of Chlamydia pneumoniae replication in human aortic smooth muscle cells by gamma interferon-induced indoleamine 2, 3-dioxygenase activity. AB - Infection with Chlamydia pneumoniae, a human respiratory pathogen, has been implicated as a potential risk factor in atherosclerosis, possibly because the pathogen can exist in a persistent form similar to that described for Chlamydia trachomatis. The present study investigated whether gamma interferon (IFN-gamma) can induce indoleamine 2,3-dioxygenase (IDO) activity in aortic smooth muscle cells, leading to a marked inhibition of C. pneumoniae growth. Our data indicate a stimulation of IDO mRNA expression and dose-dependent enzymatic activity following IFN-gamma treatment. IDO-mediated increase in tryptophan catabolism resulted in a dose-dependent marked inhibition of C. pneumoniae replication. PMID- 11035764 TI - The sequence-variable, single-copy tprK gene of Treponema pallidum Nichols strain UNC and Street strain 14 encodes heterogeneous TprK proteins. AB - Syphilis is a chronic infection with early relapses that are hypothesized to result from the emergence of phenotypic variants of Treponema pallidum. Recent studies demonstrated that TprK, a target of protective immunity, is heterogeneous in several T. pallidum strains, but not in Nichols strain Seattle (A. Centurion Lara, C. Godornes, C. Castro, W. C. Van Voorhis, and S. A. Lukehart, Infect. Immun. 68:824-831, 2000). Analysis of PCR-amplified tprK from Nichols strain UNC and Street strain 14 treponemes showed that TprK has seven regions of intrastrain heterogeneity resulting from amino acid substitutions, insertions, and deletions. In contrast, analysis of PCR-amplified tprJ showed little intrastrain or interstrain heterogeneity. Reverse transcriptase PCR analysis demonstrated that mRNA transcripts representing unique polymorphic TprK proteins are present during syphilitic infection. Southern hybridization confirmed that Nichols strain UNC and Street strain 14 each contain a single copy of tprK, indicating that intrastrain heterogeneity is due to the presence of multiple treponemal subpopulations which contain a variant form of tprK. PMID- 11035765 TI - Association of protease activity in Vibrio cholerae vaccine strains with decreases in transcellular epithelial resistance of polarized T84 intestinal epithelial cells. AB - Culture supernatants prepared from reactogenic strains of Vibrio cholerae cause a decrease in the transcellular epithelial resistance of T84 intestinal cells. This decrease correlates with the presence of hemagglutinin/protease but not with the presence of other potential accessory toxins or proteases. These data suggest a possible role for hemagglutinin/protease in reactogenicity, although other factors may also contribute. PMID- 11035766 TI - The nonfunctional allele TCRBV6S1B is strongly associated with Helicobacter pylori infection. AB - To determine genetic susceptibility factors for Helicobacter pylori infection, polymorphic T-cell receptor gene elements were investigated in 203 H. pylori infected individuals and 180 uninfected individuals (controls). H. pylori infection is highly associated with individuals homozygous for the nonfunctional TCRBV6S1B element (odds ratio = 5.9; chi(2) = 13; P = 0.00032; P value corrected for multiple comparisons [Bonferroni correction] = 0. 00063). PMID- 11035767 TI - Transmission electron microscopic demonstration of phagocytosis and intracellular processing of segmented filamentous bacteria by intestinal epithelial cells of the chick ileum. AB - Segmented filamentous bacteria (SFB) are autochthonous bacteria colonizing the ileum of many young animals by attaching to intestinal epithelial cells. These nonpathogenic bacteria strongly stimulate the mucosal immune system and induce intestinal epithelial cells to express major histocompatibility complex class II molecules. We tried to discover whether SFB are phagocytized and intracellularly processed by the host cells, which is indicative of antigen processing. The middle part of the ileum was extracted from 10- and 20-day-old broiler chicks (Gallus gallus domesticus). Samples were processed and examined by scanning and transmission electron microscopy (SEM and TEM, respectively). In SEM, no, few, medium, and dense SFB colonization levels were classified. In TEM of cells from animals with medium or dense SFB colonization levels, we could observe extracellular particles ranging from those only indenting the cell membrane to particles found in the cytoplasmatic area beyond the terminal web. These particles had a structural similarity with SFB that were floating freely in the intestinal lumen. Furthermore, we observed unlacing of the membrane and septum surrounding the extracellular particles and their incorporation into host cytoplasmatic components, which strongly suggests that these particles are phagocytized and intracellularly processed SFB. This conclusion is supported by TEM analysis of samples with no or few SFB, in which we failed to find these characteristic morphologies. The phagocytosis process described here could be an important trigger for the stimulating effect of SFB on the mucosal immune system. PMID- 11035770 TI - The magnitude distribution of declustered earthquakes in Southern California. AB - The binned distribution densities of magnitudes in both the complete and the declustered catalogs of earthquakes in the Southern California region have two significantly different branches with crossover magnitude near M = 4.8. In the case of declustered earthquakes, the b-values on the two branches differ significantly from each other by a factor of about two. The absence of self similarity across a broad range of magnitudes in the distribution of declustered earthquakes is an argument against the application of an assumption of scale independence to models of main-shock earthquake occurrence, and in turn to the use of such models to justify the assertion that earthquakes are unpredictable. The presumption of scale-independence for complete local earthquake catalogs is attributable, not to a universal process of self-organization leading to future large earthquakes, but to the universality of the process that produces aftershocks, which dominate complete catalogs. PMID- 11035769 TI - Human lung cancer and p53: the interplay between mutagenesis and selection. AB - It is an almost consensus opinion that the major carcinogenic risk of tobacco smoke is in its direct mutagenic action on DNA of cancer-related genes. The key data supposedly linking smoke-induced mutations to lung cancer were obtained from the adduct spectrum of the p53 tumor suppressor gene. Results of our analysis of p53 mutations compiled from the International Agency for Research on Cancer p53 database (April 1999 update) and from the literature point to a different causative link. Our new analytical tests focused on complementary base substitutions and showed that it is strand-specific repair of primary lesions and site-specific selection of the resultant mutations that determine the lung cancer specific hot spots of G:C to T:A transversions along the p53 gene and also their increased abundance in lung tissues as compared with smoke-inaccessible tissues. However, on each of the two strands of p53 DNA, our tests revealed no significant difference between smokers and nonsmokers, either in the frequency of different types of mutations or in the frequency of their occurrence along the p53 gene. Moreover, in both smokers and nonsmokers, there was the same frequency of lung tumors with silent p53 mutations. Accordingly, we offer here a selection-based explanation of why lung cancers with nonsilent p53 mutations are more common in smokers than in nonsmokers. We conclude that physiological stresses (not necessarily genotoxic) aggravated by smoking are the leading risk factor in the p53-associated etiology of lung cancer. PMID- 11035768 TI - Latency-associated peptide of transforming growth factor beta enhances mycobacteriocidal immunity in the lung during Mycobacterium bovis BCG infection in C57BL/6 mice. AB - Latency-associated peptide of transforming growth factor beta (TGF-beta) (LAP) was used to determine whether in vivo modulation of TGF-beta bioactivity enhanced pulmonary immunity to Mycobacterium bovis BCG infection in C57BL/6 mice. LAP decreased BCG growth in the lung and enhanced antigen-specific T-cell proliferation and gamma interferon mRNA expression. Thus, susceptibility of the lung to primary BCG infection may be partially mediated by the immunosuppressive effects of TGF-beta. PMID- 11035771 TI - Social enhancement of fitness in yellow-bellied marmots. AB - The yellow-bellied marmot (Marmota flaviventris) is a social, ground-dwelling squirrel that lives either individually or in kin groups of from two to five adult females. Philopatry and daughter recruitment lead to the formation and persistence of matrilines at habitat sites. By using 37 years of demographic data for 12 habitat sites, we could determine long-term trends in the effects of group size on two measures of fitness, survivorship and net reproductive rate, which otherwise are obscured by annual fluctuations in these measures. Both size and number of matrilines varied among sites and survivorship and net reproductive rate varied among sites and among matriline sizes. The role of social organization was explored further by examining the effect of matriline size, averaged over all years and sites, on fitness. For both survivorship and net reproductive rate the relationship with matriline size was curvilinear. Fitness increased with the increase in matriline size and then decreased in the largest groups. Decreased fitness in matrilines of four or five was associated with agonistic behavior, a large number of 2-year-old females in the social group, and reproductive suppression. There is no evidence that females acted to increase their fitness by increasing indirect fitness; i.e., by assisting relatives, but attempted to increase direct fitness. Direct fitness increased when mortality and fission of large matrilines reduced group size and the surviving females increased reproduction. PMID- 11035772 TI - Batrachotoxin alkaloids from passerine birds: a second toxic bird genus (Ifrita kowaldi) from New Guinea. AB - Batrachotoxins, including many congeners not previously described, were detected, and relative amounts were measured by using HPLC-mass spectrometry, in five species of New Guinean birds of the genus Pitohui as well as a species of a second toxic bird genus, Ifrita kowaldi. The alkaloids, identified in feathers and skin, were batrachotoxinin-A cis-crotonate (1), an allylically rearranged 16 acetate (2), which can form from 1 by sigmatropic rearrangement under basic conditions, batrachotoxinin-A and an isomer (3 and 3a, respectively), batrachotoxin (4), batrachotoxinin-A 3'-hydroxypentanoate (5), homobatrachotoxin (6), and mono- and dihydroxylated derivatives of homobatrachotoxin. The highest levels of batrachotoxins were generally present in the contour feathers of belly, breast, or legs in Pitohui dichrous, Pitohui kirhocephalus, and Ifrita kowaldi. Lesser amounts are found in head, back, tail, and wing feathers. Batrachotoxin (4) and homobatrachotoxin (6) were found only in feathers and not in skin. The levels of batrachotoxins varied widely for different populations of Pitohui and Ifrita, a result compatible with the hypothesis that these birds are sequestering toxins from a dietary source. PMID- 11035773 TI - Pinpointing when T cell costimulatory receptor CTLA-4 must be engaged to dampen diabetogenic T cells. AB - Engagement of the T cell costimulatory receptor CTLA-4 can potently down-regulate an immune response. For example, in a T cell receptor transgenic mouse model of autoimmune diabetes, CTLA-4 interactions keep pancreatic islet-reactive T cells in check, evidenced by the finding that mAb blockade of CTLA-4 rapidly provokes diabetes in animals that would not normally succumb until many months later. Interestingly, this effect is only observed early in the course of disease, before insulitis is stably entrenched. Here, we have exploited a highly synchronous and easily manipulable transfer system to determine precisely when CTLA-4 must be engaged to check the diabetogenicity of islet-reactive T cells. Our results indicate that CTLA-4 interactions during initial priming of the T cells in the pancreatic lymph nodes are not determinant. Rather, the critical interactions occur when the T cells secondarily reencounter their antigen in the target organ, the pancreatic islets. In addition, we made use of CTLA-4-deficient mice to bolster our interpretation that CTLA-4 engagement has a dampening rather than an enhancing influence on diabetes progression. PMID- 11035774 TI - Tonsillar memory B cells, latently infected with Epstein-Barr virus, express the restricted pattern of latent genes previously found only in Epstein-Barr virus associated tumors. AB - Epstein-Barr virus (EBV) establishes a life-long persistent infection in most of the human population. In the peripheral blood, EBV is restricted to memory B cells that are resting and express limited genetic information. We have proposed that these memory cells are the site of long-term persistent infection. We now show that memory cells in the tonsil express the genes for EBV nuclear antigen 1 (EBNA1) (from the Qp promoter), latent membrane protein 1 (LMP1), and LMP2a but do not express EBNA2 or the EBNA3s. This pattern of latent gene expression has only been seen previously in EBV-associated tumors such as nasopharyngeal carcinoma, Hodgkin's disease (HD), and T/NK lymphomas. Normal circulating memory B cells frequently reenter secondary lymphoid tissue, where they receive signals essential for their survival. Specifically they require signals from antigen specific T helper cells and from antigen itself. LMP1 and LMP2 are known to be able to generate these signals in a ligand-independent fashion. We suggest, therefore, that the transcription pattern we have found in latently infected, tonsillar, memory B cells is used because it allows for the expression of LMP1, LMP2a, and EBNA1 in the absence of the immunogenic and growth-promoting EBNA2 and EBNA3 molecules. LMP1 and LMP2a are produced to provide the surrogate rescue and survival signals needed to allow latently infected memory cells to persist, and EBNA1 is produced to allow replication of the viral episome. PMID- 11035775 TI - Atmospheric aerosols as prebiotic chemical reactors. AB - Aerosol particles in the atmosphere have recently been found to contain a large number of chemical elements and a high content of organic material. The latter property is explicable by an inverted micelle model. The aerosol sizes with significant atmospheric lifetimes are the same as those of single-celled organisms, and they are predicted by the interplay of aerodynamic drag, surface tension, and gravity. We propose that large populations of such aerosols could have afforded an environment, by means of their ability to concentrate molecules in a wide variety of physical conditions, for key chemical transformations in the prebiotic world. We also suggest that aerosols could have been precursors to life, since it is generally agreed that the common ancestor of terrestrial life was a single-celled organism. The early steps in some of these initial transformations should be accessible to experimental investigation. PMID- 11035777 TI - Introduction to the Japanese-American Frontiers of Science. PMID- 11035776 TI - Control of cellular cholesterol efflux by the nuclear oxysterol receptor LXR alpha. AB - LXR alpha is a nuclear receptor that has previously been shown to regulate the metabolic conversion of cholesterol to bile acids. Here we define a role for this transcription factor in the control of cellular cholesterol efflux. We demonstrate that retroviral expression of LXR alpha in NIH 3T3 fibroblasts or RAW264.7 macrophages and/or treatment of these cells with oxysterol ligands of LXR results in 7- to 30-fold induction of the mRNA encoding the putative cholesterol/phospholipid transporter ATP-binding cassette (ABC)A1. In contrast, induction of ABCA1 mRNA in response to oxysterols is attenuated in cells that constitutively express dominant-negative forms of LXR alpha or LXR beta that lack the AF2 transcriptional activation domain. We further demonstrate that expression of LXR alpha in NIH 3T3 fibroblasts and/or treatment of these cells with oxysterols is sufficient to stimulate cholesterol efflux to extracellular apolipoprotein AI. The ability of oxysterol ligands of LXR to stimulate efflux is dramatically reduced in Tangier fibroblasts, which carry a loss of function mutation in the ABCA1 gene. Taken together, these results indicate that cellular cholesterol efflux is controlled, at least in part, at the level of transcription by a nuclear receptor-signaling pathway. They suggest a model in which activation of LXRs by oxysterols in response to cellular sterol loading leads to induction of the ABCA1 transporter and the stimulation of lipid efflux to extracellular acceptors. These findings have important implications for our understanding of mammalian cholesterol homeostasis and suggest new opportunities for pharmacological regulation of cellular lipid metabolism. PMID- 11035778 TI - Quantitative transcript imaging in normal and heat-shocked Drosophila embryos by using high-density oligonucleotide arrays. AB - Embryonic development in Drosophila is characterized by an early phase during which a cellular blastoderm is formed and gastrulation takes place, and by a later postgastrulation phase in which key morphogenetic processes such as segmentation and organogenesis occur. We have focused on this later phase in embryogenesis with the goal of obtaining a comprehensive analysis of the zygotic gene expression that occurs during development under normal and altered environmental conditions. For this, a functional genomic approach to embryogenesis has been developed that uses high-density oligonucleotide arrays for large-scale detection and quantification of gene expression. These oligonucleotide arrays were used for quantitative transcript imaging of embryonically expressed genes under standard conditions and in response to heat shock. In embryos raised under standard conditions, transcripts were detected for 37% of the 1,519 identified genes represented on the arrays, and highly reproducible quantification of gene expression was achieved in all cases. Analysis of differential gene expression after heat shock revealed substantial expression level changes for known heat-shock genes and identified numerous heat shock-inducible genes. These results demonstrate that high-density oligonucleotide arrays are sensitive, efficient, and quantitative instruments for the analysis of large scale gene expression in Drosophila embryos. PMID- 11035779 TI - Coupled two-way clustering analysis of gene microarray data. AB - We present a coupled two-way clustering approach to gene microarray data analysis. The main idea is to identify subsets of the genes and samples, such that when one of these is used to cluster the other, stable and significant partitions emerge. The search for such subsets is a computationally complex task. We present an algorithm, based on iterative clustering, that performs such a search. This analysis is especially suitable for gene microarray data, where the contributions of a variety of biological mechanisms to the gene expression levels are entangled in a large body of experimental data. The method was applied to two gene microarray data sets, on colon cancer and leukemia. By identifying relevant subsets of the data and focusing on them we were able to discover partitions and correlations that were masked and hidden when the full dataset was used in the analysis. Some of these partitions have clear biological interpretation; others can serve to identify possible directions for future research. PMID- 11035780 TI - Altered selectivity in an Arabidopsis metal transporter. AB - Plants require metals for essential functions ranging from respiration to photosynthesis. These metals also contribute to the nutritional value of plants for both humans and livestock. Additionally, plants have the ability to accumulate nonessential metals such as cadmium and lead, and this ability could be harnessed to remove pollutant metals from the environment. Designing a transporter that specifically accumulates certain cations while excluding others has exciting applications in all of these areas. The Arabidopsis root membrane protein IRT1 is likely to be responsible for uptake of iron from the soil. Like other Fe(II) transporters identified to date, IRT1 transports a variety of other cations, including the essential metals zinc and manganese as well as the toxic metal cadmium. By heterologous expression in yeast, we show here that the replacement of a glutamic acid residue at position 103 in wild-type IRT1 with alanine increases the substrate specificity of the transporter by selectively eliminating its ability to transport zinc. Two other mutations, replacing the aspartic acid residues at either positions 100 or 136 with alanine, also increase IRT1 metal selectivity by eliminating transport of both iron and manganese. A number of other conserved residues in or near transmembrane domains appear to be essential for all transport function. Therefore, this study identifies at least some of the residues important for substrate selection and transport in a protein belonging to the ZIP gene family, a large transporter family found in a wide variety of organisms. PMID- 11035781 TI - Learning and memory. AB - Memory is one of the most fundamental mental processes. Neuroscientists study this process by using extremely diverse strategies. Two different approaches aimed at understanding learning and memory were introduced in this symposium. The first focuses on the roles played by synaptic plasticity, especially in long-term depression in the cerebellum in motor learning, and its regulatory mechanism. The second approach uses an elegant chick-quail transplantation system on defined brain regions to study how neural populations interact in development to form behaviorally important neural circuits and to elucidate neurobiological correlates of perceptual and motor predispositions. PMID- 11035782 TI - Extrasolar planets. AB - The first known extrasolar planet in orbit around a Sun-like star was discovered in 1995. This object, as well as over two dozen subsequently detected extrasolar planets, were all identified by observing periodic variations of the Doppler shift of light emitted by the stars to which they are bound. All of these extrasolar planets are more massive than Saturn is, and most are more massive than Jupiter. All orbit closer to their stars than do the giant planets in our Solar System, and most of those that do not orbit closer to their star than Mercury is to the Sun travel on highly elliptical paths. Prevailing theories of star and planet formation, which are based on observations of the Solar System and of young stars and their environments, predict that planets should form in orbit about most single stars. However, these models require some modifications to explain the properties of the observed extrasolar planetary systems. PMID- 11035783 TI - Aging mechanisms. AB - Aging (senescence) has long been a difficult issue to be experimentally analyzed because of stochastic processes, which contrast with the programmed events during early development. However, we have recently started to learn the molecular mechanisms that control aging. Studies of the mutant mouse, klotho, showing premature aging, raise a possibility that mammals have an "anti-aging hormone." A decrease of cell proliferation ability caused by the telomeres is also tightly linked to senescence. Frontier experimental studies of aging at the molecular level are leading to fascinating hypotheses that aging is the price we had to pay for the evolution of the sexual reproduction system that produces a variety of genetic information and complex body structures. PMID- 11035784 TI - Mantle dynamics and seismic tomography. AB - Three-dimensional imaging of the Earth's interior, called seismic tomography, has achieved breakthrough advances in the last two decades, revealing fundamental geodynamical processes throughout the Earth's mantle and core. Convective circulation of the entire mantle is taking place, with subducted oceanic lithosphere sinking into the lower mantle, overcoming the resistance to penetration provided by the phase boundary near 650-km depth that separates the upper and lower mantle. The boundary layer at the base of the mantle has been revealed to have complex structure, involving local stratification, extensive structural anisotropy, and massive regions of partial melt. The Earth's high Rayleigh number convective regime now is recognized to be much more interesting and complex than suggested by textbook cartoons, and continued advances in seismic tomography, geodynamical modeling, and high-pressure-high-temperature mineral physics will be needed to fully quantify the complex dynamics of our planet's interior. PMID- 11035785 TI - Emerging viral diseases. PMID- 11035786 TI - Inhibition of cardiac L-type calcium channels by protein kinase C phosphorylation of two sites in the N-terminal domain. AB - We have investigated the mechanism underlying the modulation of the cardiac L type Ca(2+) current by protein kinase C (PKC). Using the patch-clamp technique, we found that PKC activation by 4-alpha-phorbol 12-myristate 13-acetate (PMA) or rac-1-oleyl-2-acetylglycerol (OAG) caused a substantial reduction in Ba(2+) current through Ca(v)1.2 channels composed of alpha(1)1.2, beta(1b), and alpha(2)delta(1) subunits expressed in tsA-201 cells. In contrast, Ba(2+) current through a cloned brain isoform of the Ca(v)1.2 channel (rbC-II) was unaffected by PKC activation. Two potential sites of PKC phosphorylation are present at positions 27 and 31 in the cardiac form of Ca(v)1.2, but not in the brain form. Deletion of N-terminal residues 2-46 prevented PKC inhibition. Conversion of the threonines at positions 27 and 31 to alanine also abolished the PKC sensitivity of Ca(v)1.2. Mutant Ca(v)1.2 channels in which the threonines were converted singly to alanines were also insensitive to PKC modulation, suggesting that phosphorylation of both residues is required for PKC-dependent modulation. Consistent with this, mutating each of the threonines individually to aspartate in separate mutants restored the PKC sensitivity of Ca(v)1.2, indicating that a change in net charge by phosphorylation of both sites is responsible for inhibition. Our results define the molecular basis for inhibition of cardiac Ca(v)1.2 channels by the PKC pathway. PMID- 11035787 TI - Antimicrobial activity of MHC class I-restricted CD8+ T cells in human tuberculosis. AB - Studies of mouse models of tuberculosis (TB) infection have indicated a central role for MHC class I-restricted CD8+ T cells in protective immunity. To define antigens and epitopes of Mycobacterium tuberculosis (MTB) proteins that are presented by infected cells to CD8+ T cells, we screened 40 MTB proteins for HLA class I A*0201-binding motifs. Peptides that bound with high affinity to purified HLA molecules were subsequently analyzed for recognition by CD8+ cytotoxic T lymphocytes. We identified three epitopes recognized by CD8+ T cells from patients recovering from TB infection. Those three epitopes were derived from three different antigens: thymidylate synthase (ThyA(30-38)), RNA polymerase beta subunit (RpoB(127-135)), and a putative phosphate transport system permease protein A-1 (PstA1(75-83)). In addition, CD8+ T cell lines specific for three peptides (ThyA(30-38), PstA1(75-83), and 85B(15-23)) were generated from peripheral blood mononuclear cells of normal HLA-A*0201 donors. These CD8+ T cell lines specifically recognized MTB-infected macrophages, as demonstrated by production of IFN-gamma and lysis of the infected target cells. Finally, CD8+ cytotoxic T lymphocytes reduced the viability of the intracellular MTB, providing evidence that CD8+ T cell recognition of MHC class I-restricted epitopes of these MTB antigens can contribute to effective immunity against the pathogen. PMID- 11035788 TI - Unique progressive cleavage mechanism of HIV reverse transcriptase RNase H. AB - HIV-1 reverse transcriptase (RT) degrades the plus strand viral RNA genome while synthesizing the minus strand of DNA. Many RNA fragments, including the polypurine tracts, remain annealed to the new DNA. Several RTs are believed to bind after synthesis to degrade all RNA fragments except the polypurine tracts by a polymerization-independent mode of RNase H activity. For this latter process, we found that RT positions the RNase H active site approximately 18 nt from the 5' end of the RNA, making the primary cut. The enzyme rebinds or slides toward the 5' end of the RNA to make a secondary cut creating two products 8-9 nt long. RT then binds the new 5' end of the RNA created by the first primary or the secondary cuts to make the next primary cut. In addition, we observed another type of RNase H cleavage specificity. RT aligns the RNase H active site to the 3' end of the RNA, cutting 5 residues in. We determined the relative rates of these cuts, defining their temporal order. Results show that the first primary cut is fastest, and the secondary and 5-nt cuts occur next at similar rates. The second primary cuts appear last. Based on these results, we present a model by which RT progressively cleaves RNA fragments. PMID- 11035790 TI - SBE-TAGS: an array-based method for efficient single-nucleotide polymorphism genotyping. AB - Generating human single-nucleotide polymorphisms (SNPs) is no longer a rate limiting step for genetic studies of disease. The number of SNPs in public databases already exceeds 200,000, and the total is expected to exceed 1,000,000 within a year. Rather, progress is limited by the inability to genotype large numbers of SNPs. Current genotyping methods are suitable for studying individual loci or at most a handful at a time. Here, we describe a method for parallel genotyping of SNPs, called single base extension-tag array on glass slides, SBE TAGS. The principle is as follows. SNPs are genotyped by single base extension (SBE), using bifunctional primers carrying a unique sequence tag in addition to a locus-specific sequence. Because each locus has a distinct tag, the genotyping reactions can be performed in a highly multiplexed fashion, and the resulting product can then be "demultiplexed" by hybridization to the reverse complements of the sequence tags arrayed on a glass slide. SBE-TAGS is simple and inexpensive because of the high degree of multiplexing and the use of an easily generated, generic tag array. The method is also highly accurate: we genotyped over 100 SNPs, obtaining over 5, 000 genotypes, with approximately 99% accuracy. PMID- 11035789 TI - Insulin-like growth factor 1 regulates the location, stability, and transcriptional activity of beta-catenin. AB - The insulin-like growth factor (IGF) type 1 receptor is required for growth, transformation, and protection from apoptosis. IGFs can enhance cell migration, which is known to be influenced via regulation of the E-cadherin/beta-catenin complex. We sought to investigate whether IGF-1 modulated the interaction between E-cadherin and beta-catenin in human colorectal cancer cells. We used the C10 cell line, which we established and have previously shown to lack adenomatous polyposis coli, E-cadherin, or beta-catenin mutations. We found that IGF-1 stimulation enhanced tyrosine phosphorylation of two proteins, beta-catenin and insulin-receptor substrate 1, which formed a complex with E-cadherin. Tyrosine phosphorylation of beta-catenin was accompanied by rapid (<1 min) dissociation from E-cadherin at the plasma membrane, followed by relocation to the cellular cytoplasm. IGF-1 also enhanced the stability of beta-catenin protein. Despite this, we observed no enhancement of transcriptional activity in complex with T cell factor 4 (Tcf-4) in human embryonic kidney 293 cells treated with IGF-1 or insulin alone. IGF-1 did, however, enhance transcriptional activity in combination with lithium chloride, an inhibitor of glycogen synthase kinase 3 beta, which also stabilizes beta-catenin. In conclusion, we have shown that IGF-1 causes tyrosine phosphorylation and stabilization of beta-catenin. These effects may contribute to transformation, cell migration, and a propensity for metastasis in vivo. PMID- 11035791 TI - Deoxysugars in glycopeptide antibiotics: enzymatic synthesis of TDP-L epivancosamine in chloroeremomycin biosynthesis. AB - The 2,3,6-trideoxysugar l-epivancosamine is the terminal sugar added to the aglycone scaffold in chloroeremomycin, a member of the vancomycin family of glycopeptide antibiotics. Five proteins from the chloroeremomycin biosynthetic cluster, ORF14 and ORF23 to ORF26, have been expressed heterologously in Escherichia coli and purified to near homogeneity, and each has been characterized for an enzymatic activity. These five enzymes reconstitute the complete biosynthesis of TDP-l-epivancosamine from TDP-4-keto-6-deoxy-d-glucose. This process involves C-2 deoxygenation, C-3 amination and methylation, C-5 epimerization, and C-4 ketoreduction. Intermediates and the final product of this pathway have been identified by mass spectrometry and NMR. The pathway established here represents the complete in vitro reconstitution of an unusual sugar for an important class of antibiotics and sets the groundwork for future combinatorial biosynthesis for new bioactive compounds. PMID- 11035792 TI - Manifold anomalies in gene expression in a vineyard isolate of Saccharomyces cerevisiae revealed by DNA microarray analysis. AB - Genome-wide transcriptional profiling has important applications in evolutionary biology for assaying the extent of heterozygosity for alleles showing quantitative variation in gene expression in natural populations. We have used DNA microarray analysis to study the global pattern of transcription in a homothallic strain of Saccharomyces cerevisiae isolated from wine grapes in a Tuscan vineyard, along with the diploid progeny obtained after sporulation. The parental strain shows 2:2 segregation (heterozygosity) for three unlinked loci. One determines resistance to trifluoroleucine; another, resistance to copper sulfate; and the third is associated with a morphological phenotype observed as colonies with a ridged surface resembling a filigree. Global expression analysis of the progeny with the filigreed and smooth colony phenotypes revealed a greater than 2-fold difference in transcription for 378 genes (6% of the genome). A large number of the overexpressed genes function in pathways of amino acid biosynthesis (particularly methionine) and sulfur or nitrogen assimilation, whereas many of the underexpressed genes are amino acid permeases. These wholesale changes in amino acid metabolism segregate as a suite of traits resulting from a single gene or a small number of genes. We conclude that natural vineyard populations of S. cerevisiae can harbor alleles that cause massive alterations in the global patterns of gene expression. Hence, studies of expression variation in natural populations, without accompanying segregation analysis, may give a false picture of the number of segregating genes underlying the variation. PMID- 11035793 TI - X-ray crystal structure of an anti-Buckminsterfullerene antibody fab fragment: biomolecular recognition of C(60). AB - We have prepared a monoclonal Buckminsterfullerene specific antibody and report the sequences of its light and heavy chains. We also show, by x-ray crystallographic analysis of the Fab fragment and by model building, that the fullerene binding site is formed by the interface of the antibody light and heavy chains. Shape-complementary clustering of hydrophobic amino acids, several of which participate in putative stacking interactions with fullerene, form the binding site. Moreover, an induced fit mechanism appears to participate in the fullerene binding process. Affinity of the antibody-fullerene complex is 22 nM as measured by competitive binding. These findings should be applicable not only to the use of antibodies to assay and direct potential fullerene-based drug design but could also lead to new methodologies for the production of fullerene derivatives and nanotubes as well. PMID- 11035794 TI - A viral member of the ERV1/ALR protein family participates in a cytoplasmic pathway of disulfide bond formation. AB - Proteins of the ERV1/ALR family are encoded by all eukaryotes and cytoplasmic DNA viruses for which substantial sequence information is available. Nevertheless, the roles of these proteins are imprecisely known. Multiple alignments of ERV1/ALR proteins indicated an invariant C-X-X-C motif, but no similarity to the thioredoxin fold was revealed by secondary structure predictions. We chose a virus model to investigate the role of these proteins as thiol oxidoreductases. When cells were infected with a mutant vaccinia virus in which the E10R gene encoding an ERV1/ALR family protein was repressed, the disulfide bonds of three other viral proteins-namely, the L1R and F9L proteins and the G4L glutaredoxin were completely reduced. The same outcome occurred when Cys-43 or Cys-46, the putative redox cysteines of the E10R protein, was mutated to serine. These two cysteines were disulfide bonded during a normal virus infection but not if the synthesis of other viral late proteins was inhibited or the E10R protein was expressed by itself in uninfected cells, suggesting a requirement for an upstream viral thiol oxidoreductase. Remarkably, the cysteine-containing domains of the E10R and L1R viral membrane proteins and the glutaredoxin are in the cytoplasm, in which assembly of vaccinia virions occurs, rather than in the oxidizing environment of the endoplasmic reticulum. These data indicated a viral pathway of disulfide bond formation in which the E10R protein has a central role. By extension, the ERV1/ALR family may represent a ubiquitous class of cellular thiol oxidoreductases that interact with glutaredoxins or thioredoxins. PMID- 11035795 TI - Phosphorylation-dependent targeting of cAMP response element binding protein to the ubiquitin/proteasome pathway in hypoxia. AB - Hypoxia activates a number of gene products through degradation of the transcriptional coactivator cAMP response element binding protein (CREB). Other transcriptional regulators (e.g., beta-catenin and NF-kappa B) are controlled through phosphorylation-targeted proteasomal degradation, and thus, we hypothesized a similar degradative pathway for CREB. Differential display analysis of mRNA derived from hypoxic epithelia revealed a specific and time dependent repression of protein phosphatase 1 (PP1), a serine phosphatase important in CREB dephosphorylation. Subsequent studies identified a previously unappreciated proteasomal-targeting motif within the primary structure of CREB (DSVTDS), which functions as a substrate for PP1. Ambient hypoxia resulted in temporally sequential CREB serine phosphorylation, ubiquitination, and degradation (in vitro and in vivo). HIV-tat peptide-facilitated loading of intact epithelia with phosphopeptides corresponding to this proteasome targeting motif resulted in inhibition of CREB ubiquitination. Further studies revealed that PP1 inhibitors mimicked hypoxia-induced gene expression, whereas proteasome inhibitors reversed the hypoxic phenotype. Thus, hypoxia establishes conditions that target CREB to proteasomal degradation. These studies may provide unique insight into a general mechanism of transcriptional regulation by hypoxia. PMID- 11035796 TI - Binding sites in Escherichia coli dihydrofolate reductase communicate by modulating the conformational ensemble. AB - To explore how distal mutations affect binding sites and how binding sites in proteins communicate, an ensemble-based model of the native state was used to define the energetic connectivities between the different structural elements of Escherichia coli dihydrofolate reductase. Analysis of this model protein has allowed us to identify two important aspects of intramolecular communication. First, within a protein, pair-wise couplings exist that define the magnitude and extent to which mutational effects propagate from the point of origin. These pair wise couplings can be identified from a quantity we define as the residue specific connectivity. Second, in addition to the pair-wise energetic coupling between residues, there exists functional connectivity, which identifies energetic coupling between entire functional elements (i.e., binding sites) and the rest of the protein. Analysis of the energetic couplings provides access to the thermodynamic domain structure in dihydrofolate reductase as well as the susceptibility of the different regions of the protein to both small-scale (e.g., point mutations) and large-scale perturbations (e. g., binding ligand). The results point toward a view of allosterism and signal transduction wherein perturbations do not necessarily propagate through structure via a series of conformational distortions that extend from one active site to another. Instead, the observed behavior is a manifestation of the distribution of states in the ensemble and how the distribution is affected by the perturbation. PMID- 11035797 TI - PERK mediates cell-cycle exit during the mammalian unfolded protein response. AB - The accumulation of unfolded proteins in the endoplasmic reticulum (ER) triggers the unfolded protein response (UPR)-signaling pathway. The UPR coordinates the induction of ER chaperones with decreased protein synthesis and growth arrest in the G(1) phase of the cell cycle. Three ER transmembrane protein kinases (Ire1alpha, Ire1beta, and PERK) have been implicated as proximal effectors of the mammalian UPR. We now demonstrate that activation of PERK signals the loss of cyclin D1 during the UPR, culminating in cell-cycle arrest. Overexpression of wild-type PERK inhibited cyclin D1 synthesis in the absence of ER stress, thereby inducing a G(1) phase arrest. PERK expression was associated with increased phosphorylation of the translation elongation initiation factor 2alpha (eIF2alpha), an event previously shown to block cyclin D1 translation. Conversely, a truncated form of PERK lacking its kinase domain acted as a dominant negative when overexpressed in cells, attenuating both cyclin D1 loss and cell-cycle arrest during the UPR without compromising induction of ER chaperones. These data demonstrate that PERK serves as a critical effector of UPR induced growth arrest, linking stress in the ER to control of cell-cycle progression. PMID- 11035798 TI - Phosphorylation of murine p53 at ser-18 regulates the p53 responses to DNA damage. AB - Ser-15 of human p53 (corresponding to Ser-18 of mouse p53) is phosphorylated by ataxia-telangiectasia mutated (ATM) family kinases in response to ionizing radiation (IR) and UV light. To determine the effects of phosphorylation of endogenous murine p53 at Ser-18 on biological responses to DNA damage, we introduced a missense mutation (Ser-18 to Ala) by homologous recombination into both alleles of the endogenous p53 gene in mouse embryonic stem (ES) cells. Our analyses showed that phosphorylation of murine p53 at Ser-18 in response to IR or UV radiation was required for a full p53-mediated response to these DNA damage inducing agents. In contrast, phosphorylation of p53 at Ser-18 was not required for ATM-dependent cellular resistance after exposure to IR. Additionally, efficient acetylation of the C terminus of p53 in response to DNA damage did not require phosphorylation of murine p53 at Ser-18. PMID- 11035799 TI - Quantifying genetic and nongenetic contributions to malarial infection in a Sri Lankan population. AB - Explaining the causes of variation in the severity of malarial disease remains a major challenge in the treatment and control of malaria. Many factors are known to contribute to this variation, including parasite genetics, host genetics, acquired immunity, and exposure levels. However, the relative importance of each of these to the overall burden of malarial disease in human populations has not been assessed. Here, we have partitioned variation in the incidence of malarial infection and the clinical intensity of malarial disease in a rural population in Sri Lanka into its component causes by pedigree analysis of longitudinal data. We found that human genetics, housing, and predisposing systematic effects (e. g., sex, age, occupation, history of infections, village) each explained approximately 15% of the variation in the frequency of malarial infection. For clinical intensity of illness, 20% of the variation was explained by repeatable differences between patients, about half of which was attributable to host genetics. The other half was attributable to semipermanent differences among patients, most of which could be explained by known predisposing factors. Three percent of variation in clinical intensity was explained by housing, and an additional 7% was explained by current influences relating to infection status (e.g., parasitemia, parasite species). Genetic control of Plasmodium falciparum infections appeared to modulate the frequency and intensity of infections, whereas genetic control of Plasmodium vivax infections appeared to confer absolute susceptibility or refractoriness but not intensity of disease. Overall, the data show consistent, repeatable differences among hosts in their susceptibility to clinical disease, about half of which are attributable to host genes. PMID- 11035800 TI - Polynucleotide phosphorylase functions both as a 3' right-arrow 5' exonuclease and a poly(A) polymerase in Escherichia coli. AB - In vitro, polynucleotide phosphorylase of Escherichia coli can both synthesize RNA by using nucleotide diphosphates as precursors and exonucleolytically degrade RNA in the presence of inorganic phosphate. However, because of the high in vivo concentration of inorganic phosphate in exponentially growing cells, it has been assumed that the enzyme works exclusively as an exonuclease. Here we demonstrate that, contrary to this prediction, polynucleotide phosphorylase not only synthesizes long, highly heteropolymeric tails in vivo, but also accounts for all of the observed residual polyadenylylation in poly(A) polymerase I deficient strains. In addition, the enzyme is responsible for adding the C and U residues that are found in poly(A) tails in exponentially growing cultures of wild type E. coli. PMID- 11035801 TI - The complex ATP-Fe(2+) serves as a specific affinity cleavage reagent in ATP Mg(2+) sites of Na,K-ATPase: altered ligation of Fe(2+) (Mg(2+)) ions accompanies the E(1)-->E(2) conformational change. AB - In the presence of ascorbate/H(2)O(2), ATP-Fe(2+) or AMP-PNP-Fe(2+) complexes act as affinity cleavage reagents, mediating selective cleavage of the alpha subunit of Na,K-ATPase at high affinity ATP-Mg(2+) sites. The cleavages reveal contact points of Fe(2+) or Mg(2+) ions. In E(1) and E(1)Na conformations, two major cleavages are detected within the conserved (708)TGDGVNDSPALKK sequence (at V712 and nearby), and one (E(1)Na) or two (E(1)) minor cleavages near V440. In media containing sodium and ATP, Fe(2+) substitutes for Mg(2+) in activating phosphorylation and ATP hydrolysis. In the E(1)P conformation, cleavages are the same as in E(1). Fe(2+) is not bound tightly. By contrast, in the E(2)P conformation, the pattern is different. A major cleavage occurs near the conserved sequence (212)TGES, whereas those in TGDGVNDSPALKK are less prominent. Fe(2+) is bound very tightly. On E(2)P hydrolysis, the Fe(2+) dissociates. The results are consistent with E(1)<-->E(2) conformation-dependent movements of cytoplasmic domains and sites for P(i) and Mg(2+) ions, inferred from previous Fe cleavage experiments. Furthermore, these concepts fit well with the crystal structure of Ca-ATPase [Toyoshima, C., Nakasako, M., Nomura, H. & Ogawa, H. (2000) Nature (London) 405, 647-655]. Altered ligation of Mg(2+) ions in E(2)P may be crucial in facilitating nucleophilic attack of water on the OP bond. Mg(2+) ions may play a similar role in all P-type pumps. As affinity cleavage reagents, ATP-Fe(2+) or other nucleotide-Fe(2+) complexes could be widely used to investigate nucleotide binding proteins. PMID- 11035802 TI - Origin of mitochondria in relation to evolutionary history of eukaryotic alanyl tRNA synthetase. AB - The origin of the eukaryotic cell remains an unsolved question. Numerous experimental and phylogenetic observations support the symbiotic origin of the modern eukaryotic cell, with its nucleus and (typically) mitochondria. Incorporation of mitochondria has been proposed to precede development of the nucleus, but it is still unclear whether mitochondria were initially part of basal eukaryotes. Data on alanyl-tRNA synthetase from an early eukaryote and other sources are presented and analyzed here. These data are consistent with the notion that mitochondrial genesis did not significantly precede nucleus formation. Moreover, the data raise the possibility that diplomonads are primary amitochondriates that radiated from the eukaryotic lineage before mitochondria became fully integrated as a cellular organelle. PMID- 11035803 TI - Localizing proteins in the cell from their phylogenetic profiles. AB - We introduce a computational method for identifying subcellular locations of proteins from the phylogenetic distribution of the homologs of organellar proteins. This method is based on the observation that proteins localized to a given organelle by experiments tend to share a characteristic phylogenetic distribution of their homologs-a phylogenetic profile. Therefore any other protein can be localized by its phylogenetic profile. Application of this method to mitochondrial proteins reveals that nucleus-encoded proteins previously known to be destined for mitochondria fall into three groups: prokaryote-derived, eukaryote-derived, and organism-specific (i.e., found only in the organism under study). Prokaryote-derived mitochondrial proteins can be identified effectively by their phylogenetic profiles. In the yeast Saccharomyces cerevisiae, 361 nucleus-encoded mitochondrial proteins can be identified at 50% accuracy with 58% coverage. From these values and the proportion of conserved mitochondrial genes, it can be inferred that approximately 630 genes, or 10% of the nuclear genome, is devoted to mitochondrial function. In the worm Caenorhabditis elegans, we estimate that there are approximately 660 nucleus-encoded mitochondrial genes, or 4% of its genome, with approximately 400 of these genes contributed from the prokaryotic mitochondrial ancestor. The large fraction of organism-specific and eukaryote-derived genes suggests that mitochondria perform specialized roles absent from prokaryotic mitochondrial ancestors. We observe measurably distinct phylogenetic profiles among proteins from different subcellular compartments, allowing the general use of prokaryotic genomes in learning features of eukaryotic proteins. PMID- 11035804 TI - Suppression of angiogenesis by lentiviral delivery of PEX, a noncatalytic fragment of matrix metalloproteinase 2. AB - Modulation of the balance between pro- and antiangiogenic factors holds great promise for the treatment of a broad spectrum of human disease ranging from ischemic heart disease to cancer. This requires both the identification of angiogenic regulators and their efficient delivery to target organs. Here, we demonstrate the use of a noncatalytic fragment of matrix metalloproteinase 2 (termed PEX) delivered by lentiviral vectors in different angiogenesis models. Transduction of human endothelial cells with PEX virus suppressed endothelial invasion and formation of capillary-like structures without affecting chemotaxis in vitro. Lentiviral delivery of PEX blocked basic fibroblast growth factor induced matrix metalloproteinase 2 activation and angiogenesis on chicken chorioallantoic membranes. PEX expression also inhibited tumor-induced angiogenesis and tumor growth in a nude mouse model. Thus, our study shows that lentiviral vectors can deliver sufficient quantities of antiangiogenic substances to achieve therapeutic effects in vivo. PMID- 11035805 TI - Adenomatous polyposis coli protein contains two nuclear export signals and shuttles between the nucleus and cytoplasm. AB - Mutational inactivation of the adenomatous polyposis coli (APC) tumor suppressor initiates most hereditary and sporadic colon carcinomas. Although APC protein is located in both the cytoplasm and the nucleus, the protein domains required to maintain a predominantly cytoplasmic localization are unknown. Here, we demonstrate that nuclear export of APC is mediated by two intrinsic, leucine rich, nuclear export signals (NESs) located near the amino terminus. Each NES was able to induce the nuclear export of a fused carrier protein. Both APC NESs were independently able to interact with the Crm1 nuclear export factor and substitute for the HIV-1 Rev NES to mediate nuclear mRNA export. Both APC NESs functioned within the context of APC sequence: an amino-terminal APC peptide containing both NESs interacted with Crm1 and showed nuclear export in a heterokaryon nucleocytoplasmic shuttling assay. Also, mutation of both APC NESs resulted in the nuclear accumulation of the full-length, approximately 320-kDa APC protein, further establishing that the two intrinsic APC NESs are necessary for APC protein nuclear export. Moreover, endogenous APC accumulated in the nucleus of cells treated with the Crm1-specific nuclear export inhibitor leptomycin B. Together, these data indicate that APC is a nucleocytoplasmic shuttle protein whose predominantly cytoplasmic localization requires NES function and suggests that APC may be important for signaling between the nuclear and cytoplasmic compartments of epithelial cells. PMID- 11035806 TI - Frataxin activates mitochondrial energy conversion and oxidative phosphorylation. AB - Friedreich's ataxia (FA) is an autosomal recessive disease caused by decreased expression of the mitochondrial protein frataxin. The biological function of frataxin is unclear. The homologue of frataxin in yeast, YFH1, is required for cellular respiration and was suggested to regulate mitochondrial iron homeostasis. Patients suffering from FA exhibit decreased ATP production in skeletal muscle. We now demonstrate that overexpression of frataxin in mammalian cells causes a Ca(2+)-induced up-regulation of tricarboxylic acid cycle flux and respiration, which, in turn, leads to an increased mitochondrial membrane potential (delta psi(m)) and results in an elevated cellular ATP content. Thus, frataxin appears to be a key activator of mitochondrial energy conversion and oxidative phosphorylation. PMID- 11035807 TI - Mass spectrometry and immobilized enzymes for the screening of inhibitor libraries. AB - A technique has been developed to rapidly screen enzyme inhibitor candidates from complex mixtures, such as those created by combinatorial synthesis. Inhibitor libraries are screened by using immobilized enzyme technologies and electrospray ionization ion cyclotron resonance mass spectrometry. The library mixture is first sprayed into the mass spectrometer, and compounds are identified. The library is subsequently incubated with the immobilized enzyme of interest under the correct conditions (buffer, pH, temperature) by using an excess of enzyme to ensure a surplus of sites for ligand binding. The immobilized enzyme/inhibitor mixture is centrifuged, and an aliquot of supernatant is again analyzed by electrospray ionization mass spectrometry. Potential inhibitors are quickly identified by comparison of the spectra before and after incubation with the immobilized enzyme. Non-inhibitors show no change in ion intensity after incubation, whereas weak inhibitors exhibit a visible decrease in ion abundance. Once inhibitor candidates have been identified, the library is reinjected into the mass spectrometer, and tandem mass spectrometry is used to determine the structure of the inhibitor candidates as needed. This method has been successfully demonstrated by identifying inhibitors of the enzymes pepsin and glutathione S-transferase from a 19- and 17-component library, respectively. It is further shown that the immobilized enzyme can be recycled and reused for continuous screening of additional new libraries without adding additional enzyme. PMID- 11035808 TI - Fossilized high pressure from the Earth's deep interior: the coesite-in-diamond barometer. AB - Mineral inclusions in diamonds provide an important source of information about the composition of the continental lithosphere at depths exceeding 120-150 km, i.e., within the diamond stability field. Fossilized high pressures in coesite inclusions from a Venezuela diamond have been identified and measured by using laser Raman and synchrotron x-ray microanalytical techniques. Micro-Raman measurements on an intact inclusion of remnant vibrational band shifts give a high confining pressure of 3.62 (+/-0.18) GPa. Synchrotron single-crystal diffraction measurements of the volume compression are in accord with the Raman results and also revealed direct structural information on the state of the inclusion. In contrast to olivine and garnet inclusions, the thermoelasticity of coesite favors accurate identification of pressure preservation. Owing to the unique combination of physical properties of coesite and diamond, this "coesite in-diamond" geobarometer is virtually independent of temperature, allowing an estimation of the initial pressure of Venezuela diamond formation of 5.5 (+/-0.5) GPa. PMID- 11035809 TI - Prostate formation in a marsupial is mediated by the testicular androgen 5 alpha androstane-3 alpha,17 beta-diol. AB - Development of the male urogenital tract in mammals is mediated by testicular androgens. It has been tacitly assumed that testosterone acts through its intracellular metabolite dihydrotestosterone (DHT) to mediate this process, but levels of these androgens are not sexually dimorphic in plasma at the time of prostate development. Here we show that the 3 alpha-reduced derivative of DHT, 5 alpha-androstane-3 alpha,17 beta-diol (5 alpha-adiol), is formed in testes of tammar wallaby pouch young and is higher in male than in female plasma in this species during early sexual differentiation. Administration of 5 alpha-adiol caused formation of prostatic buds in female wallaby pouch young, and in tissue minces of urogenital sinus and urogenital tubercle radioactive 5 alpha-adiol was converted to DHT, suggesting that circulating 5 alpha-adiol acts through DHT in target tissues. We conclude that circulating 5 alpha-adiol is a key hormone in male development. PMID- 11035810 TI - Phosphorylation and inactivation of glycogen synthase kinase 3 by protein kinase A. AB - Glycogen synthase kinase 3 (GSK-3) is implicated in multiple biological processes including metabolism, gene expression, cell fate determination, proliferation, and survival. GSK-3 activity is inhibited through phosphorylation of serine 21 in GSK-3 alpha and serine 9 in GSK-3 beta. These serine residues of GSK-3 have been previously identified as targets of protein kinase B (PKB/Akt), a serine/threonine kinase located downstream of phosphatidylinositol 3-kinase. Here, we show that serine 21 in GSK-3 alpha and serine 9 in GSK-3 beta are also physiological substrates of cAMP-dependent protein kinase A. Protein kinase A physically associates with, phosphorylates, and inactivates both isoforms of GSK 3. The results indicate that depending on the stimulatory context, the activity of GSK-3 can be modulated either by growth factors that work through the phosphatidylinositol 3-kinase-protein kinase B cascade or by hormonal stimulation of G protein-coupled receptors that link to changes in intracellular cAMP levels. PMID- 11035811 TI - Cytochrome c binding to Apaf-1: the effects of dATP and ionic strength. AB - In the apoptosis pathway in mammals, cytochrome c and dATP are critical cofactors in the activation of caspase 9 by Apaf-1. Until now, the detailed sequence of events in which these cofactors interact has been unclear. Here, we show through fluorescence polarization experiments that cytochrome c can bind to Apaf-1 in the absence of dATP; when dATP is added to the cytochrome c.Apaf-1 complex, further assembly occurs to produce the apoptosome. These findings, along with the discovery that the exposed heme edge of cytochrome c is involved in the cytochrome c.Apaf-1 interaction, are confirmed through enhanced chemiluminescence visualization of native PAGE gels and through acrylamide fluorescence quenching experiments. We also report here that the cytochrome c.Apaf-1 interaction depends highly on ionic strength, indicating that there is a strong electrostatic interaction between the two proteins. PMID- 11035812 TI - Regulation of the antioxidant response element by protein kinase C-mediated phosphorylation of NF-E2-related factor 2. AB - A coordinated cellular response to oxidative stress occurs in part through transcriptional regulation via a cis-acting sequence known as the antioxidant response element (ARE). NF-E2-related factor 2 (Nrf2), a member of the Cap'n'Collar family of basic region-leucine zipper (bZIP) transcription factors, has been implicated as an essential component of an ARE-binding transcriptional complex, but the signaling pathway leading to its activation has remained unclear. Using a reporter gene assay, we found that ARE-directed transcription was activated by phorbol 12-myristate 13-acetate (PMA), but completely suppressed by staurosporine and Ro-32-0432, selective inhibitors of protein kinase C (PKC). Immunocytochemistry and subcellular fractionation revealed that PMA, like tert butylhydroquinone (tBHQ), promoted the nuclear localization of Nrf2, a process that was blocked by staurosporine or Ro-32-0432. We showed that Nrf2, a previously unidentified kinase target, was phosphorylated in HepG2 cells. PMA transiently activated Nrf2 phosphorylation, whereas the addition of tBHQ or beta naphthoflavone (betaNF) led to a persistent stimulation, which was abolished by staurosporine, but not by U0126 and SB203580, respective inhibitors of MEK and p38 kinases. Purified Nrf2 was phosphorylated in vitro by the catalytic subunit of PKC, or by PKC immunoprecipitated from cell lysates. Significantly, PKC precipitated from tBHQ- or betaNF-treated cells showed enhanced activity against Nrf2. These findings indicate an important role of the PKC pathway in the ARE mediated gene expression, and suggest that PKC-directed phosphorylation of Nrf2 may be a critical event for the nuclear translocation of this transcription factor in response to oxidative stress. PMID- 11035813 TI - The semaphorin receptor plexin-B1 specifically interacts with active Rac in a ligand-dependent manner. AB - Semaphorin molecules serve as axon guidance signals that regulate the navigation of neuronal growth cones. Semaphorins have also been implicated in other biological processes, including the immune response. Plexins, acting either alone or in complex with neuropilins, have recently been identified as functional semaphorin receptors. However, the mechanisms of signal transduction by plexins remain largely unknown. We have demonstrated a direct interaction between plexin B1 and activated Rac. Rac specifically interacts with the cytosolic domain of plexin-B1, but not with that of plexin-A3 or -C1. Neither RhoA nor Cdc42 interacts with plexin-B1, indicating that the Rac/plexin-B1 interaction is highly specific. The binding of GTP and the integrity of the Rac effector domain are required for the interaction with plexin-B1. Furthermore, we have identified that a Cdc42/Rac interactive binding (CRIB) motif in the cytosolic domain of plexin-B1 is essential for its interaction with active Rac. We have also observed that the semaphorin CD100, a ligand for plexin-B1, stimulates the interaction between plexin-B1 and active Rac. Our results support a model by which activated Rac plays a role in mediating semaphorin signals, resulting in reorganization of actin cytoskeletal structure. PMID- 11035814 TI - Detection of mutations in transgenic fish carrying a bacteriophage lambda cII transgene target. AB - To address the dual needs for improved methods to assess potential health risks associated with chemical exposure in aquatic environments and for new models for in vivo mutagenesis studies, we developed transgenic fish that carry multiple copies of a bacteriophage lambda vector that harbors the cII gene as a mutational target. We adapted a forward mutation assay, originally developed for lambda transgenic rodents, to recover cII mutants efficiently from fish genomic DNA by lambda in vitro packaging. After infecting and plating phage on a hfl- bacterial host, cII mutants were detected under selective conditions. We demonstrated that many fundamental features of mutation analyses based on lambda transgenic rodents are shared by transgenic fish. Spontaneous mutant frequencies, ranging from 4.3 x 10(-5) in liver, 2.9 x 10(-5) in whole fish, to 1.8 x 10(-5) in testes, were comparable to ranges in lambda transgenic rodents. Treatment with ethylnitrosourea resulted in concentration-dependent, tissue-specific, and time dependent mutation inductions consistent with known mechanisms of action. Frequencies of mutants in liver increased insignificantly 5 days after ethylnitrosourea exposure, but increased 3.5-, 5.7- and 6. 7-fold above background at 15, 20, and 30 days, respectively. Mutants were induced 5-fold in testes at 5 days, attaining a peak 10-fold induction 15 days after treatment. Spontaneous and induced mutational spectra in the fish were also consistent with those of lambda transgenic rodent models. Our results demonstrate the feasibility of in vivo mutation analyses using transgenic fish and illustrate the potential value of fish as important comparative animal models. PMID- 11035816 TI - Turner's syndrome: a growing concern. PMID- 11035815 TI - The pachytene checkpoint prevents accumulation and phosphorylation of the meiosis specific transcription factor Ndt80. AB - In budding yeast, many mutants defective in meiotic recombination and chromosome synapsis undergo checkpoint-mediated arrest at the pachytene stage of meiotic prophase. We recovered the NDT80 gene in a screen for genes whose overexpression bypasses the pachytene checkpoint. Ndt80 is a meiosis-specific transcription factor that promotes expression of genes required for exit from pachytene and entry into meiosis I. Herein, we show that the Ndt80 protein accumulates and is extensively phosphorylated during meiosis in wild type but not in cells arrested at the pachytene checkpoint. Our results indicate that inhibition of Ndt80 activity is one mechanism used to achieve pachytene arrest. PMID- 11035817 TI - Pieces of the puzzle: diabetes and the structure and function of the heart and blood vessels. PMID- 11035818 TI - Atopy, autoimmunity, and the T(H)1/T(H)2 balance. PMID- 11035819 TI - Effect of inhaled corticosteroids on the hypothalamic-pituitary-adrenal axis and growth in children. PMID- 11035820 TI - Delayed diagnoses of Turner's syndrome: proposed guidelines for change. AB - OBJECTIVE: To measure the delays in diagnosis of Turner's syndrome (TS) and to propose strategies for earlier screening and diagnosis. METHODS: The medical records of 81 girls with TS were reviewed for age at diagnosis, reason(s) for karyotype analysis, and clinical features including growth failure. Delay in diagnosis was calculated as equal to age at diagnosis for children born with lymphedema and/or 2 or more of the following dysmorphic features: webbed neck, nail dysplasia, high palate, and short fourth metacarpal. For all others, delay in diagnosis was calculated as the difference between the age at which height fell below the 5th percentile and the age at which the diagnosis of TS was made. RESULTS: Lymphedema was the key to diagnosis in 97% of the girls diagnosed with TS in infancy, and short stature was the key to diagnosis for 82% of the girls diagnosed in childhood or adolescence. For girls diagnosed in childhood or adolescence, the delay in diagnosis averaged 7.7 +/- 5.4 years. Many had dysmorphic features and/or a history of lymphedema at birth, and diagnosis was made an average of 5.3 years after patients had fallen below the 5th percentile for height. By the time of diagnosis, patients were very short, averaging -2.9 SD in height. CONCLUSIONS: The diagnosis of TS is often delayed. We recommend karyotype analysis for all girls with unexplained short stature, delayed puberty, webbed neck, lymphedema, or coarctation of the aorta. Furthermore, karyotype analysis should be strongly considered for those who remain above the 5th percentile for height but have 2 or more features of TS, including high palate, nail dysplasia, short fourth metacarpal, and strabismus. PMID- 11035821 TI - Longitudinal analysis of growth over the first 3 years of life in Turner's syndrome. AB - OBJECTIVE: To evaluate longitudinal growth in Turner's syndrome (TS) over the first 3 years of life. METHODS: Growth of 47 patients with TS was compared with that of 40 age-matched control girls by using an analysis according to the Infancy-Childhood-Puberty and bi-exponential models. RESULTS: A mean of 1.2 SDs were lost before birth and a total of 3.0 SDs were lost by age 3 years. According to the Infancy-Childhood-Puberty model, intrauterine growth retardation contributed -1.24 SDs, a 5-month delay in childhood growth spurt contributed 0.96 SDs, and slow childhood growth contributed an additional -0.8 SDs by age 3 years. The bi-exponential analysis disclosed a quasi-linear first exponent and a confining second exponent, which merged at age 18 months in control subjects and 24 months in patients with TS. The first exponent confers an average annual growth rate of 8.4 cm/y in control subjects and 6.7 cm/y in patients with TS. CONCLUSIONS: Intrauterine growth retardation and the initial 3 years of life contribute most of the deficit in the final height of patients with TS. These data provide a reference of standards for longitudinal growth in patients with TS at age 3 months to 3 years. PMID- 11035822 TI - Carotid artery distensibility and cardiac function in adolescents with type 1 diabetes. AB - OBJECTIVE: To determine the presence and correlates of early heart and blood vessel dysfunction in adolescents with type 1 diabetes mellitus (DM) of relatively short duration. STUDY DESIGN: A total of 33 patients with DM (20 male, mean age 15.8 +/- 1.3 years, mean DM duration 9.3 +/- 3.9 years) and 16 healthy subjects in a nondiabetic control group (7 male, mean age 17.4 +/- 1.7 years) underwent (1) ultrasonography of the right carotid artery to assess distensibility, compliance, and intimal-medial thickness (IMT), (2) echocardiographic assessment of systolic and diastolic ventricular function, (3) lipid profile and hemoglobin A(1c), and (4) overnight timed urine collections for albumin excretion rate. RESULTS: Ultrasonography showed significantly lower carotid artery distensibility in the DM group (38.5 +/- 8.2 x 10(-3) vs 46.5 +/- 11.7 x 10(-3)/kPa, P =.01) but no difference in compliance (14.0 +/- 3.4 x 10(-7) vs 15.8 +/- 2.9 x 10(-7)m(2)/kPa, P =.08) or IMT (0.061 +/- 0.013 vs 0.060 +/- 0.014 cm, P =.77). Left ventricular (LV) end-diastolic diameter, LV posterior wall thickness, end-systolic wall stress, shortening fraction, ejection fraction, LV mass, and diastolic function were similar in both groups. Total cholesterol, low density lipoprotein, high density lipoprotein, triglycerides, and blood pressure were also similar. The median albumin excretion rate was 4.8 microg/min in the DM group (range 1.1 to 19.2) and 3.0 microg/min in the control group (range 1.4 to 5.8) (P =.03). Hemoglobin A(1c) correlated inversely with both distensibility (r = -.43, P =.02) and compliance (r = -.39, P =.032). CONCLUSIONS: This study indicates that early changes in macrovascular function, namely lower carotid artery distensibility, may precede abnormalities in cardiac function or in arterial IMT in adolescents with short duration type 1 DM. It also supports a relationship between hyperglycemia and carotid artery dysfunction. PMID- 11035823 TI - Decreased prevalence of atopic diseases in children with diabetes. The EURODIAB Substudy 2 Study Group. AB - OBJECTIVE: To test the hypothesis that atopic diseases in early life are associated with a reduced risk (protection) for the development of type 1 diabetes in childhood. STUDY DESIGN: European centers (n = 8) with access to population-based type 1 diabetes registries (>90% degree of ascertainment) participated in a case control study focusing on early exposures and risk factors for type 1 diabetes. Altogether, data from 1028 members of a case group and 2744 members of a control group corresponding to 85.4% eligible members of the case group and 76.1% of the control group were analyzed. Information in this study was collected regarding atopic diseases (atopic eczema, allergic rhinoconjunctivitis, and asthma). RESULTS: Atopic disease and asthma particularly are associated with significant reductions in risk of childhood type 1 diabetes. The risk reduction associated with asthma was observed reasonably consistently among the 8 study centers, which represent a wide range of diabetes incidence. Risk reductions associated with all 3 expressions of atopy were particularly marked in children whose type 1 diabetes was diagnosed in the 10- to 14-year age group. CONCLUSION: These findings indicate that atopic conditions may be protective against the development of type 1 diabetes and are consistent with the immunologic concept of T(H)1 (type 1 diabetes) and T(H)2 (atopy) diseases being mutually exclusive. PMID- 11035824 TI - The association of atopic dermatitis in infancy with immunoglobulin E food sensitization. AB - OBJECTIVE: To prospectively investigate the association of high levels of immunoglobulin E (IgE) sensitization to foods and the presence of atopic dermatitis (judged by reported topical steroid use during the first 16 months of life) in a birth cohort of 620 Australian children "at risk" of allergic disease because of family history. RESULTS: A total of 559 of the children in the cohort were fully evaluated, and the cumulative prevalence of atopic dermatitis was 24%. More children in the cohort who had atopic dermatitis had strongly positive skin test results (> or = 4+, histamine equivalent units, > or = approximately 6-mm wheal), consistent with IgE food sensitization to either cow's milk, egg, or peanut at 6 months (22% vs 5%, chi(2) = 35; P < 10(-6)) and at 12 months (36% vs 11%, chi(2) = 41; P < 10(-6)) than those without atopic dermatitis. The calculated attributable risk percent for IgE food sensitization as a cause of atopic dermatitis was 65% and 64% at these times. In a separate group of infants with severe atopic dermatitis, the equivalent rates of IgE food sensitization at 6 months was 83% and at 12 months, 65%. CONCLUSION: IgE food sensitization is a major risk factor for the presence of atopic dermatitis in infancy. PMID- 11035825 TI - Allergic gastroenteropathy in preterm infants. AB - OBJECTIVES: To determine the clinical presentation, histopathologic features, and outcome of biopsy-proven allergic gastroenteropathy (AGE) in preterm infants. We hypothesized that AGE is a more frequent cause of gastrointestinal disease in this population than previously suspected. STUDY DESIGN: The retrospective portion of the study, from 1992 to 1997, included preterm infants <37 weeks' gestation who underwent biopsy because of suspected AGE. The prospective portion, from January to December 1998, included 20 infants undergoing endoscopy and biopsy because of suspected AGE. RESULTS: Twenty-five infants (12 retrospective/13 prospective) with mean gestational age of 29 weeks at birth and mean postnatal age at diagnosis of 78 days were diagnosed with AGE. Three clinical patterns of presentation were noted: group 1, gastroesophageal reflux disease (n = 5); group 2, non-specific feeding intolerance (n = 8); and group 3, lower gastrointestinal bleeding (n = 12). Ten patients had negative biopsy findings (3 retrospective/7 prospective) and had clinical features indistinguishable from those of groups 1 and 2. Patients in group 3 were most likely to have positive biopsy findings (12 of 12). Fifteen patients responded to a casein hydrolysate formula, and 10 patients required an amino acid-based formula. Patients with AGE who had eosinophilic infiltration and villous atrophy took longer to recover than those with eosinophilic infiltration alone (P <.03). Subsequently, most have tolerated formula challenges and are currently tolerating cow's milk. CONCLUSIONS: AGE may be an under-recognized cause of gastrointestinal symptoms in preterm infants. Confirmation with endoscopy and biopsy can be done safely and provides the basis for appropriate dietary management. PMID- 11035826 TI - Deficiency of humoral immunity in cartilage-hair hypoplasia. AB - OBJECTIVE: Cartilage-hair hypoplasia (CHH), a metaphyseal chondrodysplasia, is usually associated with impaired cellular immunity. This study evaluates humoral immunity in patients with CHH. METHODS: The concentrations of immunoglobulins G, A, and M (IgG, IgA, and IgM) and IgG subclasses were studied in 20 patients. Data for 5 additional patients with recurrent infections were retrospectively reviewed. RESULTS: Seven of the prospectively evaluated patients (35%) had defective humoral immunity. Three patients had IgA deficiency. Four patients had IgG2 deficiency, accompanied by IgA deficiency, IgG4 deficiency, or both in 3 patients. IgG4 was low in most patients. Increased infections were usually associated with supranormal IgG and IgG1 and subnormal IgA, IgG2, or IgG4 concentrations. One retrospectively reviewed patient had severe hypogammaglobulinemia, and 3 had multiple IgG subclass deficiencies. CONCLUSIONS: Humoral immunity is impaired in CHH and contributes to the increased susceptibility to infections. PMID- 11035827 TI - White coat hypertension in children with elevated casual blood pressure. AB - OBJECTIVE: We reviewed our experience using ambulatory blood pressure monitoring (ABPM) in children referred to a hypertension clinic to determine the frequency of pediatric white coat hypertension (WCH). STUDY DESIGN: WCH was defined by 3 different diagnostic criteria: (1) mean 24-hour blood pressure (BP) less than Task Force-defined 95th percentile, (2) mean 24-hour BP less than 95th percentile from pediatric normative ABPM data, and (3) mean 24-hour BP less than ABPM 95th percentile and BP load (percentage of BP readings during 24-hour period exceeding the 95th percentile) less than 25%. RESULTS: Clinic BP values were available in 67 otherwise healthy children who underwent ABPM; 51 had confirmed clinic hypertension by Task Force criteria. WCH frequency in these 51 patients with the stated criteria was 53%, 45%, and 22%, respectively. Elevated BP load was found in 52% (12/23) of patients with normal mean BP. CONCLUSION: These results suggest that many children referred for casual BP elevation have WCH even by strict diagnostic criteria. ABPM may help differentiate WCH from persistent hypertension, thereby avoiding unnecessary diagnostic evaluation and identifying children most likely to benefit from early intervention. PMID- 11035828 TI - Discontinuation of enterostomy tube feeding by behavioral treatment in early childhood: a randomized controlled trial. AB - OBJECTIVE: To determine whether behavior therapy was more effective than nutritional therapy in obviating the need for enteral feeding in infants with resistance to feeding. STUDY DESIGN: Sixty-four children aged 4 to 36 months who were tube fed for at least 1 month and had resistance to feeding were randomly assigned to either behavioral or nutritional interventions (32 per group). For 7 consecutive weeks subjects and their primary feeders attended a weekly clinic with 1 of 2 dietitians followed by 4 follow-up visits. The nutritional intervention provided structured schedules and routines to stimulate the hunger/satiety cycle. The behavioral intervention provided the same schedules and routines plus behavioral therapy (extinction). The primary outcome measure was the proportion of successes, defined as infants no longer requiring tube feeding at the third follow-up visit in each group (4(1/2) months after start of trial). The decision to discontinue tube feeding was made by an independent observer who used criteria defined before the study commencement. RESULTS: Fifteen (47%) of 32 subjects in the behavioral group versus none in the nutritional group were successes (P <.001). CONCLUSION: Behavior therapy is more efficacious in eliminating the need for tube feeding than nutritional counseling alone. PMID- 11035829 TI - Effects of long-chain polyunsaturated fatty acid supplementation on fatty acid status and visual function in treated children with hyperphenylalaninemia. AB - BACKGROUND: Children with phenylalanine-hydroxylase deficiency (type-I hyperphenylalaninemia, HPA) follow a low-phenylalanine diet, severely restricted in animal foods and long-chain polyunsaturated fatty acids (LCPUFA). Consequently, they have a poor LCPUFA status, particularly for docosahexaenoic acid (DHA). DHA is relevant to visual and neural development. OBJECTIVE: To investigate the effects of a 12-month supplementation with LCPUFA in a double blind, placebo-controlled trial in treated children with HPA. STUDY DESIGN: Twenty children with well-controlled HPA were randomly allocated to receive either a fat supplement (supplying 26% as fatty acids including DHA, 8%) or a placebo. The fatty acid composition of erythrocyte lipids and the visual evoked potentials were measured at baseline and after 12 months of supplementation. Reference data were obtained from healthy children of comparable age. RESULTS: At baseline children with HPA had a poorer DHA status and prolonged P100 wave latencies than the reference group. At the end of the trial the LCPUFA group showed a significant increase in DHA levels of erythrocyte lipids. In the LCPUFA group P100 wave latency decreased and was negatively associated with the DHA changes. CONCLUSIONS: A balanced dietary supplementation with LCPUFA in children with HPA is associated with an increase of the DHA pool and improved visual function. PMID- 11035830 TI - Short stature in competitive prepubertal and early pubertal male gymnasts: the result of selection bias or intense training? AB - OBJECTIVES: To determine whether high-volume, high-impact physical training in prepubertal and early pubertal male gymnasts is associated with reduced statural and segmental growth and reduced serum insulin-like growth factor-I (IGF-I) and increased cortisol (C) levels. STUDY DESIGN: Height, sitting height, leg length, and segmental lengths (humerus, radius, femur, and tibia) and breadths (biacromial and bi-iliac), diet, serum IGF-I, testosterone, and C were measured in competitive male gymnasts and normoactive children (Tanner stage < or = 2) every 3 to 4 months over an 18-month period. RESULTS: At baseline, gymnasts (n = 31) were 0.7 years older than members of the control group (P <.05, n = 50) but were no different in terms of biologic maturity. Age-adjusted z scores showed that the gymnasts were shorter than members of the control group (-0.5 +/- 0.2 SD, P <.05) because of reduced leg length (-0.8 +/- 0.2 SD, P <.001) but not sitting height. Segmental lengths and bi-iliac breadth age-adjusted z scores were also reduced in the gymnasts (P ranging <.05 to <.001). No difference was detected for serum IGF-I or C. After 18 months of follow-up, no differences were found for rates of change in height, sitting height or leg length, segmental lengths, IGF-I, or C between those gymnasts and control subjects who remained prepubertal and early pubertal (gymnasts n = 18; control group n = 35). However, the magnitudes of baseline differences in anthropometric measures (z scores) persisted throughout the study. CONCLUSION: Short stature in these competitive male gymnasts was due to a reduced leg length but not sitting height. The lack of a difference in growth rates, IGF-I, and diet over the 18-month period indicates that the short stature reported in male gymnasts is due to selection bias rather than gymnastics training. PMID- 11035831 TI - Bronchoalveolar lavage cellular composition in acute asthma and acute bronchiolitis. AB - OBJECTIVE: To compare cellular inflammation in the airways between acute bronchiolitis and asthma. STUDY DESIGN: Using a bronchoalveolar lavage with flexible bronchoscopy procedure, we investigated the cellular constituents of BAL fluid in children with acute exacerbation of asthma (n = 18) and infants with acute bronchiolitis caused by respiratory syncytial virus (n = 20). These results were compared with those of healthy control subjects (n = 14). RESULTS: Total lavage fluid recovered was similar in all groups. The total cell numbers were highest in the bronchiolitis group. The BAL cellular profile in the asthma group was characterized by a higher median (interquartile range) ratio of eosinophils (2.4% [1.6%-9.5%]; P <.01) than in the bronchiolitis group (0% [0%-0%]) or the control group (0% [0%-0%]). Neutrophil ratio was higher in the bronchiolitis group (40.0% [26.5%-50.0%]; P <.01), with no difference found between the asthma group (3.3% [2.0%-7.9%]) and the control group (2.0% [0.8%-5.5%]). CONCLUSIONS: Asthma and acute bronchiolitis are characterized by an elevated cellular percentage of eosinophils and neutrophils, respectively, in bronchoalveolar lavage fluid. PMID- 11035832 TI - Feeding efficiency and respiratory integration in infants with acute viral bronchiolitis. AB - OBJECTIVE: To examine the effects of bronchiolitis on feeding efficiency and respiratory integration. STUDY DESIGN: We studied 21 infants with bronchiolitis and 21 bottle-fed healthy infants who formed a comparison group. Repeat evaluations of half the bronchiolitis group were performed during recovery. During each feeding study we measured the duration and frequency of sucking, the frequency of single and multiple swallows, the respiratory rate, the postswallow respiratory direction, and the suck and swallow volumes. RESULTS: The infants with bronchiolitis devoted significantly less time to sucking than their healthy peers (P <.05), and the mean suck volume was reduced. Although the frequency of swallowing was slightly higher, the volume of milk consumed per swallow was almost half the amount consumed by the comparison group (P <.01). Coordination of breathing with swallowing was also less effective (P <.01). CONCLUSION: Although most aspects of feeding are less efficient during periods of respiratory illness, others are preserved or recover rapidly. Coordination of breathing during feeding is also significantly impaired. PMID- 11035833 TI - The interaction of prematurity with genetic and environmental influences on cognitive development in twins. AB - OBJECTIVE: To investigate how the degree of prematurity interacts with genetic and environmental influences in their effect on verbal and nonverbal cognitive development. STUDY DESIGN: The target sample consisted of more than 2000 pairs of twins born in England and Wales in 1994. At 24 months, measures of verbal and non verbal cognitive development were obtained from the twins' parents. The sample was divided into 3 groups according to degree of prematurity: very preterm or high-risk (<32 weeks), moderately preterm or medium-risk (32-33 weeks), and mildly preterm/term or low-risk (>34 weeks). Quantitative genetic analyses were used to assess the contributions of genetic and environmental influences on vocabulary and cognitive development. RESULTS: The results indicated gene environment interactions. For the high-risk group, genetic effects on both verbal and non-verbal cognitive ability were completely overshadowed by shared environmental factors, whereas for both medium- and low-risk groups, additive genetic effects explained 18% to 33% of the variance. CONCLUSIONS: Our findings indicate that genetic factors are not responsible for cognitive outcomes of very preterm infants and suggest that early environmental influences appear to affect verbal and non-verbal cognitive development at 2 years of age. PMID- 11035835 TI - Unbound bilirubin associated with kernicterus: a historical approach. AB - OBJECTIVE: To determine the unbound bilirubin concentration (UBC) associated with kernicterus with the use of clinical data from clusters of kernicterus after sulfisoxazole and benzyl alcohol administration. DESIGN: Sulfisoxazole at 12 mg/dL and benzoate at 10 mmol/L are associated with kernicterus at total bilirubins near 12 and 10 mg/dL, respectively. The concurrent UBC was estimated by first measuring the drug-induced increases in UBC in plasma and artificial sera (peroxidase-diazo method). The increases were then applied to baseline UBC, determined by linear regression analysis of binding data (peroxidase method) from 86 newborns, at total bilirubins of 12 mg/dL for sulfisoxazole and 10 mg/dL for benzoate. Sensitivity and specificity were determined with existing data. RESULTS: Sulfisoxazole and benzoate increased UBC in artificial sera 2.1-fold and 4.1-fold, respectively, and in plasma (sulfisoxazole) 2.4-fold. Benzoate would increase baseline UBC from 0.29 to 1.19 microg/dL and sulfisoxazole from 0.36 to 0.86 microg/dL. The sensitivity and specificity of a UBC of 0.86 microg/dL for predicting kernicterus are 79% and 92% and for 1.19 microg/dL, 50% and 98%, respectively. CONCLUSION: Historic data predict that the unbound bilirubin above which kernicterus becomes likely lies between 0.86 and 1.19 microg/dL, in good agreement with existing information. PMID- 11035834 TI - Health status development in a cohort of preterm children. AB - OBJECTIVE: To determine the impact of preterm birth on health status (HS) development at the ages of 5 and 10 years in a cohort of children born before term. SAMPLE: Six hundred eighty-eight children, born in 1983 with a gestational age of <32 weeks and a birth weight of <1500 g. DESIGN: Prospectively collected HS variables, obtained from the parents, were analyzed in a longitudinal perspective by using principal component analyses. RESULTS: One third of the sample had minor to severe HS problems at both ages of measurement. One third had problems on one assessment only. The remainder of the sample had no HS problems at either age. The analyses grouped the HS variables into 3 combinations. Problems in basic functioning, such as mobility or speech, decreased with age. Negative moods substantially increased, and concentration problems increased slightly. Specifically at risk were preterm born children with handicaps, boys, and children who were small for gestational age. CONCLUSION: According to the parents, one third of the cohort had no HS problems at either age. The pattern of HS problems of the preterm born children changed between 5 and 10 years of age. PMID- 11035836 TI - The role of swallowing during active sleep in the clearance of reflux in term and preterm infants. AB - OBJECTIVES: To investigate swallowing and peristalsis in sleep and during gastroesophageal reflux (GER) in both healthy term and preterm infants at term equivalent age. STUDY DESIGN: Multichannel recordings were made in 12 healthy term and 11 preterm infants, under the same conditions, after feeding. Sleep state, cardiorespiratory variables, esophageal pH, and pharyngeal swallowing and peristalsis were measured. GER was defined as pH <4 for > or = 15 seconds, and swallows were classified as pharyngeal only, primary peristalsis (propagated, dropped, interrupted), or secondary peristalsis. RESULTS: Spontaneous swallowing rate was not significantly different between term and preterm infants and was sleep state-related, occurring in active sleep but rarely in quiet sleep. In response to acid GER, term infants significantly increased pharyngeal swallowing from a median of 0.7 (25th-75th interquartile range, 0.5-0.9) to 1.7 (1.0-3.0) swallows/min and secondary peristalsis from a median of 0.5 (25th-75th interquartile range, 0.3-0.8) to 1.1 (0.8-2.0) waves/min (P <.05). In contrast, the preterm infants demonstrated a significantly higher proportion of fully propagated peristaltic swallows compared with the term infants (53% and 27%, respectively) (P <.05). CONCLUSION: The occurrence of swallowing is sleep state related. In active sleep, term infants clear GER by increasing swallowing and secondary peristalsis, whereas preterm infants at term equivalent age clear GER by increasing propagated peristalsis. This method of clearance would explain the mechanism by which preterm infants have significantly shorter episodes of reflux than term infants. PMID- 11035837 TI - Lead exposure from blood transfusion to premature infants. AB - OBJECTIVE: The objective of this study was to determine the exposure of premature infants to lead from blood transfusions. STUDY DESIGN: Blood lead concentrations were determined for 19 very premature infants at the time of admission, at 4 weeks of life, and before and after transfusions and in the donor packed red blood cells of 79 transfusions. RESULTS: The number of transfusions per patient was 4. 2 +/- 2.8 (mean +/- SD) with 15.7 +/- 1.9 mL/kg packed red blood cells for a lead dose of 1.56 +/- 1.77 microg/dL. The total dose of lead from these transfusions over the 4-week period was 4.0 +/- 2.8 microg/kg (range, 0.9-10.6 microg/kg). Increases in post-transfusion blood lead concentration were linear with doses higher than 1.5 microg/dL. Packed red blood cells with a blood lead concentration of > or = 5 microg/dL resulted in an elevated post-transfusion blood lead concentration in some infants. CONCLUSIONS: The lead exposure to these infants through blood transfusion exceeds the acceptable daily intake values for lead and may result in unacceptably high post-transfusion blood lead concentrations. Use of packed red blood cells with lead concentrations <3.3 microg/dL is one cost-effective means to reduce exposure. PMID- 11035839 TI - Purpura fulminans caused by group A beta-hemolytic Streptococcus sepsis. PMID- 11035838 TI - Lead exposure and motor functioning in 4(1/2)-year-old children: the Yugoslavia prospective study. AB - OBJECTIVE: To investigate associations between lead exposure and early motor development. STUDY DESIGN: We conducted standardized assessments of motor function (Bruininks-Oseretsky Test of Motor Proficiency and Beery Developmental Test of Visual-Motor Integration) at age 54 months in 283 children whose mothers were recruited in pregnancy from a smelter town and a non-lead-exposed town in Yugoslavia and who have been monitored twice yearly since birth. Blood lead concentration (BPb) was summarized in a measure reflecting the average of the child's semiannual serial log BPbs through 54 months. RESULTS: Multiple regression showed that taken together, anthropometric measures (birth weight, body mass index) and markers of a stimulating and organized home life (HOME scale, parental education and intelligence, availability of siblings) explained a significant 10% to 18% of the variance in motor functioning. Beyond these contributions, BPb was significantly associated with poorer fine motor and visual motor function but was unrelated to gross motor coordination. CONCLUSIONS: Modest associations between early lead exposure and fine motor and visual motor functioning appear even after statistical adjustment is done for other contributors to motor development. Associations with BPb are specific to these areas of motor skill; gross motor development was unaffected. PMID- 11035840 TI - Symptomatic lead poisoning in infancy: a prospective case analysis. AB - This report of a case of symptomatic lead poisoning in infancy reinforces the need for continued vigilance in screening and the application of effective therapies to prevent serious physiologic, neurocognitive, and behavioral sequelae. Furthermore, this case illustrates the efficacy of repeated courses of outpatient succimer therapy in limiting a rebound in blood lead concentrations. PMID- 11035841 TI - Living-related liver transplantation for neonatal-onset propionic acidemia. AB - We report a child with neonatal-onset propionic acidemia treated with living related liver transplantation. Despite minimal improvement in the levels of circulating propionyl CoA metabolites, hyperammonemia was corrected, and no episode of metabolic decompensation was experienced after the transplantation was performed. Natural protein intake could be increased from 0.5 g/kg per day to 2 g/kg per day. Anemia was corrected, and the growth rate and mental development improved significantly. PMID- 11035842 TI - Resolution of hypoxemia in a liver transplant recipient after ligation of a portosystemic shunt. AB - This report describes the unique development of pulmonary vascular dilatation and hypoxemia associated with a portosystemic shunt in a pediatric liver transplant recipient. Ligation of the shunt resulted in resolution of hypoxemia. The outcome suggests that hepatic venous return to the pulmonary circulation is important in maintaining normal pulmonary vascular caliber. PMID- 11035843 TI - Psoriatic eruption in Kawasaki disease. AB - We describe 10 patients who developed a psoriatic skin eruption during either the acute or convalescent phase of Kawasaki disease. The skin eruption was pustular in 3 patients, but more typical psoriasiform skin lesions were seen in the remaining 7 patients. No patient has yet developed chronic psoriasis. PMID- 11035844 TI - Scleral melanocytosis and oculodermal melanocytosis (nevus of Ota) in Chinese children. AB - Two thousand nine hundred fourteen Chinese children (1510 males and 1404 females) were examined for the presence of scleral melanocytosis (SM) and oculodermal melanocytosis in a cross-sectional prevalence survey. SM was found in 4.9% of boys and 4.1% of girls under the age of 1 year. The peak prevalence was at 6 years of age, when 44.6% of boys and 46.6% of girls were affected. At 18 years of age, only 11.1% of boys and 13.2% of girls had SM. The overall prevalence, regardless of age, was 27.6% in boys and 27. 1% in girls. The condition was bilateral in 78% of cases. The medial superior quadrant was the most frequently affected site, and the lateral inferior quadrant was the least frequently affected site. Oculodermal melanocytosis occurred only in one patient; the pigmentation affected the left side of the face and the ipsilateral sclera. PMID- 11035845 TI - Hyperfunctioning malignant thyroid nodule in an 11-year-old girl: pathologic and molecular studies. AB - We identified a papillary carcinoma in an 11-year-old girl with a hyperfunctioning thyroid nodule. A met453thr mutation in TSHR was found in the nodule but not in normal thyroid tissue or in leukocytes. This case documents that this activating mutation is associated with neoplasia. PMID- 11035846 TI - Delivery in breech presentation--a cause of hemorrhoids in a newborn. PMID- 11035847 TI - Acute chest syndrome of sickle cell disease. PMID- 11035849 TI - Reply PMID- 11035848 TI - Acute chest syndrome of sickle cell disease. PMID- 11035851 TI - Reply PMID- 11035850 TI - Interpreting risks of steroids for preterm infants. PMID- 11035852 TI - FDA public health advisory: risk of electromagnetic interference with medical telemetry systems. PMID- 11035853 TI - What can one vote do? PMID- 11035854 TI - Preventing bicycle-related head trauma in children. AB - Although the majority of bicycle-related injuries and deaths could be prevented by the use of bicycle helmets, rates of helmet use in the United States remain well below Healthy People 2000's goal of 50% usage. Educational efforts to improve usage rates often fail to produce significant changes, in part because a child's understanding of risk plays only a small role in his or her decision to wear a helmet. To address the need for more effective injury prevention techniques, the authors propose a modification of the Health Belief Model, which is used by injury control experts to explain the various cognitive, social, and environmental factors that influence preventative health behavior. By incorporating the behavioral theory of self-efficacy in the structure of the Health Belief Model, trauma care providers and injury prevention specialists will be better able to design successful injury prevention programs that address key variables in health-related decision making. PMID- 11035855 TI - Emergency department thoracotomy: nursing implications for pediatric cases. AB - Cardiac arrest in the pediatric patient is an infrequent event. Although an emergency department thoracotomy is a potentially lifesaving procedure, it should be used in only a small, select group of patients. A literature review was conducted to determine the indications, surgical techniques, emergency procedures, and nursing responsibilities associated with an emergency department thoracotomy. PMID- 11035857 TI - Strategies for teaching safety education to children with special needs. AB - A nurse describes how a student community health project was used to develop a unique safety education program. Seat belt safety was presented to a group of school-age children who had special physical and emotional needs. The author lists the overall principles of conducting an educational session and provides examples of how those principles were applied with this student group. PMID- 11035858 TI - Lisfranc's or tarsometatarsal fracture-dislocation. AB - Lisfranc's injuries are one of the most commonly missed fractures in a multitrauma patient. Failure to recognize and treat such an injury can lead to acute (e.g., compartment syndrome of the foot) and chronic problems (e.g., arthritis). A list of reminders is included to help with the recognition and treatment of Lisfranc's injuries (Table 1). PMID- 11035859 TI - Holiday safety Web sites. PMID- 11035860 TI - White coat hypertension in children. AB - Ambulatory blood pressure monitoring (ABPM) has become more widely used in the assessment of elevated blood pressure in children. The accurate diagnosis of white coat hypertension (WCH) is particularly important in children because detection of elevated blood pressure often results in expensive and invasive diagnostic procedures to detect underlying disease. Recent normative pediatric data have both enhanced our ability to interpret ABPM results in pediatric patients and increased awareness that children suffer from WCH as has already been reported in adults. The few studies of WCH in children report a prevalence ranging from 44-88%, depending on the choice of threshold values for normalcy. When persistent hypertension is confirmed by three blood pressure measurements on three different occasions, ABPM should be performed as part of the initial evaluation. If hypertension is confirmed by ABPM, further evaluation should be tailored to the individual patient depending on the age, severity of hypertension, associated risk factors, and presence of end-organ injury. PMID- 11035861 TI - Is nurse-measured blood pressure a valid substitute for ambulatory blood pressure monitoring? AB - BACKGROUND: Because ambulatory blood pressure monitoring (ABPM) is not available everywhere, the objective of the study was to determine whether nurse-measured blood pressure could be an acceptable substitute to ABPM. METHODS: We analyzed the data of 2385 consecutive patients referred to our hypertension clinic for the performance of ABPM. Before ambulatory monitoring was performed, a nurse-measured BP was obtained three times using a Y-tube connecting the sphygmomanometer and the recorder. We compared the mean value of the three nurse-measured blood pressures with that of the 12h daytime ambulatory monitoring, considered as the reference. RESULTS: The difference between the nurse-measured and the ambulatory blood pressure was small but statistically significant, indicating that nurse measured blood pressure tends to overestimate both diastolic and systolic blood pressure. The difference between the nurse blood pressure and ABPM was greater among treated hypertensive patients than untreated patients. To diagnose hypertension, defined as a blood pressure of over 140/90mmHg by ABPM, the positive predictive value of the nurse blood pressure was 0.81 and the negative predictive value 0.63. However, these predictive values could be improved with less stringent cut-off values of blood pressure. Thus, for a diastolic blood pressure above 100mmHg, the positive predictive value of nurse blood pressure was 0.55 and the negative predictive value 0.91. These figures were relatively similar for previously treated and untreated patients. CONCLUSION: Nurse blood pressure is less accurate than ABPM in diagnosing hypertension, defined as a blood pressure of over 140/90mmHg. It could, however, be an acceptable substitute, especially to exclude people who do not need to be treated, in situations where lower resources require a less rigorous definition of hypertension. PMID- 11035862 TI - Impact of ambulatory blood pressure monitoring on the management of hypertension in children. AB - BACKGROUND: Ambulatory blood pressure monitoring (ABPM) is widely utilized for the evaluation and management of hypertension in adults but has not been routinely used in the care of hypertensive children. DESIGN: To examine the potential impact that the routine use of ABPM might have on the evaluation and management of hypertension in children, we reviewed our early experience with this technique, comparing management decisions based on ABPM with those based on casual blood pressure readings. METHODS: Twenty children (4-17 years old) underwent ABPM for either the initial evaluation of suspected hypertension or the ongoing management of known hypertension. ABPM studies were considered abnormal if over 40% of both systolic and/or diastolic blood pressure readings were above the 95th percentile for the child's age and gender. RESULTS: Of 13 children studied for suspected hypertension, nine had abnormal ABPM studies and were treated, but four had normal results and were therefore not treated. Of seven children with known hypertension studied by ABPM to assess their blood pressure control further, three had normal results and no management changes were made, but four had abnormal studies, prompting a change in therapy or further diagnostic testing. Overall, eight out of 20 ABPM studies led to the initiation of further diagnostic evaluation or a change in planned or ongoing therapy. CONCLUSIONS: We conclude that ABPM has the potential to become an important tool in the evaluation and management of childhood hypertension, and suggest that more widespread use of this technique in children is appropriate. PMID- 11035863 TI - Reliability of nocturnal blood pressure dipping. AB - BACKGROUND: Increasing evidence documents the fact that individuals whose blood pressure drops or 'dips' relatively little at night have a higher risk of numerous cardiovascular illnesses. OBJECTIVE: To examine the reliability of various measures of nocturnal blood pressure dipping. METHODS: This study examined 17 individuals with ambulatory blood pressure monitoring on three 24 h recordings while they pursued a schedule similar to that of in-patients on a clinical research unit. Nocturnal dipping of blood pressure was scored three ways: as the drop in blood pressure between 10 p.m. and 6 a.m. ('clocktime'), as the drop in blood pressure tailored to each individual's reported bedtime ('bedtime'), and as the drop in blood pressure accompanying polysomnographically verified sleep ('sleeptime'). RESULTS: Adequate reliability was obtained for all three measures of dipping. There was, in general, a significant correlation across testing occasions (P<0.05). The correlation coefficient ranged from 0.5 to 0.8, depending on which criterion of dipping was selected and whether the endpoint was systolic blood pressure, diastolic blood pressure, or mean arterial blood pressure. CONCLUSIONS: The reliability of systolic blood pressure dipping was somewhat lower than that of diastolic or mean arterial blood pressure dipping. Dipping appears to be a reliable construct. While no one definition of dipping was demonstrably better than another, the most sensible definition of dipping would allow some adjustment for defining 'night' on the basis of each individual's idiosyncratic bed time. PMID- 11035865 TI - Validation of A&D UA-767 device for the self-measurement of blood pressure. AB - BACKGROUND: The validation of self-measurement devices has been recommended. Automatic monitor A&D UA-767 (A&D Company, Ltd, Tokyo, Japan) is well known and widely used, but not tested according to the Association for Advancement of Medical Instrumentation (AAMI) and British Hypertension Society (BHS) recommendations. OBJECTIVE: To perform a clinical validation for use by adults of the A&D UA-767 device according to the criteria of the AAMI and a modified BHS protocol. METHODS: The test concerned 101 subjects (57 men and 44 women) aged 15 85 years with arm circumferences of 22-39 cm, a systolic blood pressure (SBP) range of 89-206 mmHg, and a diastolic blood pressure (DBP) range of 53-122 mmHg. For each subject, three readings of the UA-767 were compared with simultaneous auscultatory measurements by two trained independent observers who used a mercury manometer and dual stethoscope. The results were graded according to the BHS (1990 and 1993) and AAMI recommendations. RESULTS: Observers showed close agreement, with mean differences of 1.1+/-2.4 mmHg for SBP and -0.7+/-2.0 mmHg for DBP. The proportion of values agreeing to within 5, 10 and 15 mmHg were 93, 100, and 100% for SBP and 97, 100, and 100% for DBP for the two observers. The sphygmomanometer measurements were 132+/-24/79+/-14 mmHg (mean+/-SD). The average difference between the mercury sphygmomanometer and A&D UA-767 readings for SBP and DBP were, respectively, -0.4+/-5.4 and -0.4+/-4. 8 mmHg. The proportion of values agreeing to within 5,10, and 15 mmHg were 82, 94, and 98% for SBP and 80, 95, and 98% for DBP for the observers and device (A/A grade for BHS). CONCLUSIONS: For an adult population, the A&D UA-767 device for the self measurement of blood pressure satisfied the AAMI criteria, achieved a BHS grade of A/A and can therefore be recommended for monitoring blood pressure in home and clinical conditions by patients with mild-to-moderate arterial hypertension. PMID- 11035864 TI - Results of antihypertensive treatment by primary and secondary care physicians as assessed by ambulatory blood pressure monitoring. AB - BACKGROUND: We present data from a cross-sectional study on consecutive non randomized drug-treated mild-to-moderate essential hypertensives, whose blood pressure was ambulatorily monitored for 24 h to evaluate the presence of adequate control. DESIGN: Primary and secondary care physicians were invited to send to our clinic drug-treated patients with essential hypertension (JNC VI stages 1-2) to undergo 24-h ambulatory blood pressure monitoring (ABPM) while continuing their prescribed medications. METHODS: The 436 enrolled patients (255 males, 181 females, age 61+/-11 years) were left on their therapeutic regime: monotherapy in 208 patients (47. 7%) and combination therapy in 228 patients (52.3%). All the patients were divided into two care groups: primary care, 238 patients (54.6%) and secondary care, 198 patients (45.4%). A mean daytime blood pressure < or =135/85 mmHg was chosen as a definition of adequate blood pressure control. RESULTS: Adequate blood pressure control was found in 196/436 total patients (45%); 112/238 patients in primary care (47%) and 84/198 patients in secondary care (42.4%) (P=NS); 94/208 patients (45.2%) in monotherapy and 102/228 patients (44.7%) in combination therapy (P=NS); 125/255 male patients (49%) and 71/181 female patients (39.2%) (P=0.0428). In the logistic regression model, female sex was associated with a higher risk of inadequate blood pressure control of about 50%. CONCLUSIONS: Adequate blood pressure control, as assessed by ABPM, is not different in the two settings of family doctor's office and specialist's clinic and is predicted by male gender. The figures of adequate blood pressure control remind us of the rule of halves, regardless of treatment regimes and medications. PMID- 11035866 TI - A validation of the Mobil O Graph (version 12) ambulatory blood pressure monitor. AB - OBJECTIVE: To assess the clinical accuracy of the Mobil O Graph (version 12) ambulatory blood pressure monitor in an adult population. METHODS: The accuracy of the device was assessed by predefined criteria (British Hypertension Society, BHS) in 85 subjects recruited from the patients and staff in a teaching hospital. A series of same-arm sequential blood pressure measurements were taken: first two observers taking simultaneous mercury readings, followed by a reading with the Mobil O Graph ambulatory monitor. A total of seven readings were taken from each subject in the sitting position. The data were then analysed according to the BHS protocol and the criteria of the Association for the Advancement of Medical Instrumentation (AAMI). RESULTS: The Mobil O Graph ambulatory monitor fulfilled the criteria of the BHS protocol, achieving a grade B for systolic blood pressure (SBP) and a grade A for diastolic blood pressure (DBP). The mean differences were -2+/-8 mmHg for SBP and -2+/-7 mmHg for DBP. The device therefore also passed the AAMI standard (the mean to be within 5+/-8 mmHg). CONCLUSION: The Mobil O Graph ambulatory monitor performed in a satisfactory manner according to the BHS and the AAMI criteria and can therefore be recommended for clinical use in the general population. PMID- 11035867 TI - Clinical evaluation of the efficacy of the Braun PrecisionSensor oscillometric wrist blood pressure monitor for use on adults versus auscultation as defined by ANSI/AAMI SP10-1992. AB - OBJECTIVE: To evaluate the overall performance of a new oscillometric wrist blood pressure monitor (Braun PrecisionSensor, Braun GmbH, Kronberg, Germany) as defined by the ANSI/AAMI SP10-1992 guidelines, and to analyze the data for the optimized selection of the algorithm that derives the blood pressure values from the oscillometric blood pressure curves. METHODS: The clinical trial was a multi center, open, within-subject evaluation. Repeated sequential blood pressure measurements were taken on the left wrist using the Braun PrecisionSensor, and on the left upper arm using a T-tube stethoscope and a mercury sphygmomanometer as a standard auscultatory blood pressure measurement device. The reported results are based on a sample of 86 adult male and female subjects (57% female, 43% male). Three sets of measurements with each of both devices were evaluated for each individual. RESULTS: Close agreement was obtained between both observers in compliance with the ANSI/AAMI SP10-1992 guidelines. The mean values of the differences between the Braun PrecisionSensor and the mercury sphygmomanometer were 0.1mmHg for systolic and 1.9mmHg for diastolic blood pressure. The standard deviations were 7.1mmHg for systolic and 7.0mmHg for diastolic blood pressure. The use of an optimized algorithm improved the accuracy of the Braun PrecisionSensor, after which the standard deviations were 6.1mmHg for systolic and 6.3mmHg for diastolic blood pressure. CONCLUSIONS: The Braun PrecisionSensor satisfies the Association for the Advancement of Medical Instrumentation's efficacy and safety criteria for both systolic and diastolic blood pressures with both algorithms analyzed. PMID- 11035869 TI - JAMA 100 years ago: alleged insanity in the army PMID- 11035868 TI - A piece of my mind: five miles from tomorrow. PMID- 11035870 TI - 2000 Albert Lasker Medical Research Awards. PMID- 11035871 TI - Genital herpes vaccine shows limited promise. PMID- 11035872 TI - Health agencies update: promising HIV vaccine target PMID- 11035873 TI - Health agencies update: cold comfort PMID- 11035874 TI - Health agencies update: toolbox for drug abuse treatment PMID- 11035875 TI - Health agencies update: colon cancer screening PMID- 11035876 TI - Statin drugs and the risk of fracture. PMID- 11035877 TI - Statin drugs and the risk of fracture. PMID- 11035878 TI - Statin drugs and the risk of fracture PMID- 11035879 TI - Problems in pharmacoeconomic analyses. PMID- 11035881 TI - Problems in pharmacoeconomic analyses PMID- 11035880 TI - Problems in pharmacoeconomic analyses. PMID- 11035882 TI - Problems in pharmacoeconomic analyses PMID- 11035883 TI - Clinical diagnosis of carpal tunnel syndrome. PMID- 11035884 TI - Clinical diagnosis of carpal tunnel syndrome. PMID- 11035885 TI - Clinical diagnosis of carpal tunnel syndrome PMID- 11035886 TI - Outcomes of ancrod in acute ischemic stroke. PMID- 11035887 TI - Outcomes of ancrod in acute ischemic stroke. Independent Data and Safety Monitoring Board for ESTAT. Steering Committee for ESTAT. European Stroke Treatment with Ancrod Trial. PMID- 11035888 TI - Outcomes of ancrod in acute ischemic stroke PMID- 11035890 TI - Regional brain volume abnormalities and long-term cognitive outcome in preterm infants. AB - CONTEXT: Preterm infants have a high prevalence of long-term cognitive and behavioral disturbances. However, it is not known whether the stresses associated with premature birth disrupt regionally specific brain maturation or whether abnormalities in brain structure contribute to cognitive deficits. OBJECTIVE: To determine whether regional brain volumes differ between term and preterm children and to examine the association of regional brain volumes in prematurely born children with long-term cognitive outcomes. DESIGN AND SETTING: Case-control study conducted in 1998 and 1999 at 2 US university medical schools. PARTICIPANTS: A consecutive sample of 25 eight-year-old preterm children recruited from a longitudinal follow-up study of preterm infants and 39 term control children who were recruited from the community and who were comparable with the preterm children in age, sex, maternal education, and minority status. MAIN OUTCOME MEASURES: Volumes of cortical subdivisions, ventricular system, cerebellum, basal ganglia, corpus callosum, amygdala, and hippocampus, derived from structural magnetic resonance imaging scans and compared between preterm and term children; correlations of regional brain volumes with cognitive measures (at age 8 years) and perinatal variables among preterm children. RESULTS: Regional cortical volumes were significantly smaller in the preterm children, most prominently in sensorimotor regions (difference: left, 14.6%; right, 14.3% [P<.001 for both]) but also in premotor (left, 11.2%; right, 12.6% [P<.001 for both]), midtemporal (left, 7.4% [P =.01]; right, 10.2% [P<.001]), parieto occipital (left, 7.9% [P =.01]; right, 7.4% [P =.005]), and subgenual (left, 8.9% [P =.03]; right, 11.7% [P =.01]) cortices. Preterm children's brain volumes were significantly larger (by 105. 7%-271.6%) in the occipital and temporal horns of the ventricles (P<. 001 for all) and smaller in the cerebellum (6.7%; P =.02), basal ganglia (11.4%-13.8%; P90%) found the procedure to be acceptable. CONCLUSIONS: Our results suggest that vaginal misoprostol, 800 microg, can be used from 1 to 3 days after mifepristone, 200 mg, for early medical abortion, and need not be administered strictly 48 hours after mifepristone. JAMA. 2000;284:1948-1953. PMID- 11035892 TI - Cost-effectiveness of screening for colorectal cancer in the general population. AB - CONTEXT: A recent expert panel recommended that persons at average risk of colorectal cancer (CRC) begin screening for CRC at age 50 years using 1 of several strategies. However, many aspects of different CRC screening strategies remain uncertain. OBJECTIVE: To assess the consequences, costs, and cost effectiveness of CRC screening in average-risk individuals. DESIGN: Cost effectiveness analysis from a societal perspective using a Markov model. SUBJECTS: Hypothetical subjects representative of the 50-year-old US population at average risk for CRC. SETTING: Simulated clinical practice in the United States. MAIN OUTCOME MEASURES: Discounted lifetime costs, life expectancy, and incremental cost-effectiveness (CE) ratio, compared used 22 different CRC screening strategies, including those recommended by the expert panel. RESULTS: In 1 base-case analysis, compliance was assumed to be 60% with the initial screen and 80% with follow-up or surveillance colonoscopy. The most effective strategy for white men was annual rehydrated fecal occult blood testing (FOBT) plus sigmoidoscopy (followed by colonoscopy if either a low- or high-risk polyp was found) every 5 years from age 50 to 85 years, which resulted in a 60% reduction in cancer incidence and an 80% reduction in CRC mortality compared with no screening, and an incremental CE ratio of $92,900 per year of life gained compared with annual unrehydrated FOBT plus sigmoidoscopy every 5 years. In a base-case analysis in which compliance with screening and follow-up is assumed to be 100%, screening more often than every 10 years was prohibitively expensive; annual rehydrated FOBT plus sigmoidoscopy every 5 years had an incremental CE ratio of $489,900 per life-year gained compared with the same strategy every 10 years. Other strategies recommended by the expert panel were either less effective or cost more per year of life gained than the alternatives. Colonoscopy every 10 years was less effective than the combination of annual FOBT plus sigmoidoscopy every 5 years. However, a single colonoscopy at age 55 years achieves nearly half of the reduction in CRC mortality obtainable with colonoscopy every 10 years. Because of increased life expectancy among white women and increased cancer mortality among blacks, CRC screening was even more cost-effective in these groups than in white men. CONCLUSIONS: Screening for CRC, even in the setting of imperfect compliance, significantly reduces CRC mortality at costs comparable to other cancer screening procedures. However, compliance rates significantly affect the incremental CE ratios. In this model of CRC, 60% compliance with an every 5-year schedule of screening was roughly equivalent to 100% compliance with an every 10-year schedule. Mathematical modeling used to inform clinical guidelines needs to take into account expected compliance rates. JAMA. 2000;284:1954-1961. PMID- 11035893 TI - Use of the Web for medical information by a gastroenterology clinic population. AB - CONTEXT: Surveys have shown that 60 million persons in the United States searched for health information online in 1998. However, lack of sampling from a clinic population limits the generalizability of these surveys to clinical practice. OBJECTIVES: To determine gastroenterology patients' access to and use of the Web as a medical information resource, to identify for what information patients search, and to determine how often physicians recommend that patients search the Web. DESIGN, SETTING, AND PARTICIPANTS: Cross-sectional survey of 1006 gastroenterology outpatients in Durham, NC, and Rockford, Ill, conducted in August 1999. MAIN OUTCOME MEASURES: Patient characteristics and education level, access to the Web, use of the Web as a medical information resource, search methods, and plans for future Web use. RESULTS: A total of 924 patients (92%) completed the questionnaire. Median age was 53 years, 41% were men, and the median education level was having completed some college. Fifty percent (462/924) reported having access to the Web. Of the 462 with access, 235 (51%) had searched the Web for medical information within the previous 12 months. Therefore, 25.5% of all patients surveyed had searched the Web for medical information within the previous year. Sixty percent of patients intended to use the Web as a medical information resource in the future. Only 35 (4%) of 825 had ever been referred to the Web by a physician. CONCLUSIONS: In this clinic setting, more than one quarter of gastroenterology outpatients reported having obtained medical information from the Web within the previous year. More than two thirds of patients stated they would use the Web as a medical information resource in the future. JAMA. 2000;284:1962-1964. PMID- 11035894 TI - A proposed national policy on health care workers living with HIV/AIDS and other blood-borne pathogens. AB - In 1991, scientific uncertainty about the risk of transmission of human immunodeficiency virus or hepatitis B virus (hepatitis B e antigen [HBeAg] positive) led the Centers for Disease Control and Prevention to recommend that infected health care workers (HCWs) be reviewed by an expert panel and inform patients of their serologic status before engaging in exposure-prone procedures. The data demonstrate that risks of transmission in the health care setting are exceedingly low, suggesting that the national policy should be reformed. Implementation of the current national policy at the local level poses significant human rights burdens on HCWs, but does not improve patient safety. A new national policy should focus on the management of the workplace environment and injury prevention by creating a program to prevent blood-borne pathogen transmission; by encouraging infected HCWs to promote their own health and well being; by discontinuing expert review panels and special restrictions for exposure-prone procedures, which stigmatize HCWs; by discontinuing mandatory disclosure of a HCW's infection status in low-level risk procedures; and by imposing practice restrictions to avert significant risks to patients. Inclusion of these principles would achieve high levels of patient safety without discrimination and invasion of privacy. JAMA. 2000;284:1965-1970. PMID- 11035895 TI - Dopamine agonists in early therapy for Parkinson disease: promise and problems. PMID- 11035896 TI - Perinatal brain injury in preterm infants and later neurobehavioral function. PMID- 11035897 TI - Patient access to information on clinicians infected with blood-borne pathogens. PMID- 11035898 TI - Screening strategies for early detection of lung cancer: the time is now. PMID- 11035899 TI - Routine screening for lung cancer?: Maybe someday, but not yet. PMID- 11035900 TI - Fractures in the elderly: epidemiology and demography. AB - Osteoporosis and osteoporosis-related fractures are a major source of both morbidity and cost in the elderly, the fractures that are most commonly associated with osteoporosis being those of the hip, the distal forearm and the vertebrae, although it is believed that most other fractures occurring in the elderly are also related to osteoporosis. In this review, the incidence of all types of fracture is described based on the available literature, and the foreseeable trends resulting from demographic changes are discussed. Emphasis is given to the epidemiology of hip fracture since this is the most serious consequence of osteoporosis. Hip fractures occur all over the world, most currently occurring in Western countries, mainly Europe and the USA, but it is expected that there will be a large increase in the number of hip fractures in other countries because of demographic changes. The incidence of hip fractures increases exponentially with age, resulting in a 1-year incidence of 1% in women aged 80 in Western countries. Most hip fractures occur in women, but this is again partly due to demography, because of the longer life expectancy of women. Wrist fractures occur more often in women and do not show the same increase with age as hip fractures. The incidence reaches a plateau at age 60-70. Vertebral fractures show a modest increase with age and are again more common in women than men. The incidence of all other fractures increases modestly with age PMID- 11035901 TI - Pathogenesis of osteoporosis. AB - Osteoporosis is a very common disorder affecting millions of post-menopausal women and men of various ages. Although the disease is manifested by painful fractures of the spine, hip or radius, the underlying pathogenesis is complex and multifactorial. One of the strongest predictors of future osteoporotic fractures is low bone mineral density. The determinants of adult bone density include the rate of bone acquisition during adolescence and the absolute loss of bone during the six decades of adult life. Recent studies have clarified how bone mass is acquired during the early teen years in both boys and girls. Genetic factors play an enormous role in defining the height of acquisition of bone mass; however, these factors also interact with environmental and hormonal determinants. Many more studies have focused on adult bone loss. Disorders in bone remodelling result in an imbalance in bone turnover, favouring resorption over formation. Systemic factors such as oestrogen deprivation and parathyroid hormone strongly activate remodelling and can, in several circumstances, lead to imbalances in the remodelling cycle. The molecular cues that couple bone formation to resorption have recently been elucidated, and those factors may themselves become therapeutic targets for future treatment regimens to prevent osteoporosis and its resultant fractures. PMID- 11035902 TI - The use of bone densitometry in clinical practice. AB - Bone densitometry is an established method for the assessment of osteoporosis as, according to the definition of osteoporosis, an accurate determination of the level of bone mass is central to the diagnostic assessment of osteoporosis. The diversity of different bone densitometry techniques, however, needs to be acknowledged. The World Health Organization criteria of osteoporosis should not be used for peripheral measurements, and their application to subject groups other than white women is still controversial. A large variety of bone densitometry and quantitative ultrasound techniques can be used for fracture risk assessment. Their results should be interpreted in the context of other clinical examinations and can then be used in making treatment decisions. For monitoring purposes, the ratio of response rate and long-term precision error determines longitudinal sensitivity. For all of these applications, careful quality assurance procedures need to be implemented. If applied in a responsible fashion, bone densitometry represents a powerful approach that is indispensable for the assessment of osteoporosis. PMID- 11035903 TI - The role of bone turnover markers and risk factors in the assessment of osteoporosis and fracture risk. AB - The clinical evaluation of osteoporosis in individual patients involves confirmation of the diagnosis, the investigation of secondary causes of osteoporosis and the evaluation of subsequent fracture risk. Optimum clinical assessment involves bone mineral densitometry with the treatment thresholds modified by clinical risk factors for individual patients. Bone turnover markers and clinical risk factors can be used to identify patients at risk of osteoporotic fracture and those who have secondary osteoporosis. Risk assessment should involve the evaluation of absolute rather than relative risk. Further work is required to improve the integration of clinical risk factors, bone turnover markers and bone densitometry into appropriate models to enable the assessment of the absolute risk of fracture for individual patients. PMID- 11035904 TI - Selection of individuals for prevention of fractures due to bone fragility. AB - Most patients with fractures go untreated because of the lack of awareness of osteoporosis. Treatment is indicated for women and men with osteoporosis and women and men with fractures with either osteoporosis or osteopenia because (a) fractures increase morbidity and mortality, (b) the burden of fractures is increasing because longevity is increasing, and (c) bone loss accelerates, rather than decelerates in old age. The indication for drug therapy is less clear in women or men with osteopenia because drugs have not been proved to reduce fracture risk in this group. There is no evidence that treating individuals with only risk factors reduces the fracture rate. Screening has not been shown to reduce the burden of fractures. Altering the bone mineral density by a few percent in the population is likely to reduce the number of fractures, but how this can be achieved is unknown. The rigorously investigated drugs reducing the spine fracture rate are alendronate, raloxifene and risedronate. Calcium and vitamin D reduce hip fractures in nursing home residents but not community dwellers. In the community, only alendronate and risedronate have been reported to reduce hip fractures in randomized trials. The evidence for hormone replacement therapy is less satisfactory. It is likely to reduce the number of spinal fractures, but its role in hip fracture prevention is uncertain. Only alendronate has been reported to reduce spine fractures in men with osteoporosis. Evidence for the use of other drugs (calcitonin, fluoride, anabolic steroids and active vitamin D metabolites) in women or men is insufficient to justify their use. PMID- 11035905 TI - Prevention of osteoporotic fractures in post-menopausal women. AB - A number of pharmacological interventions are now available for the prevention of osteoporotic fractures in post-menopausal women. These include hormone replacement therapy, bisphosphonates, raloxifene, calcitonin, calcitriol and combined calcium and vitamin D. Factors influencing the positioning of these agents in clinical practice include their efficacy in preventing fractures at both the spine and the hip, tolerability, side-effects, cost and, in the case of raloxifene and hormone replacement therapy, the extra-skeletal risks and benefits of long-term treatment. The rates of onset and offset of the treatment effect are also important considerations; the observations that relatively short-term intervention produces a significant reduction in fracture risk in women with established osteoporosis, that treatment benefits are greatest in those with low bone mineral density and that the beneficial skeletal effects are not maintained after the withdrawal of treatment have resulted in a shift from long-term preventive strategies towards the targeting of high-risk individuals for intervention. PMID- 11035906 TI - Non-pharmacological interventions. AB - The aim of non-pharmacological intervention for osteoporosis is to prevent, treat or alleviate the consequences of osteoporosis, the main one of which is fracture. Non-pharmacological interventions consist of a wide spectrum of treatment modalities to decrease pain, correct postural change, improve mobility, enable the patient to follow a normal social life and prevent (further) fracture. An exercise programme can increase bone mass in adolescents and adults, but in the elderly its main emphasis should be on improving muscle strength and balance in order to decrease the risk of falls. Physiotherapy is commonly prescribed to mobilize the patient after a fracture, to decrease muscle spasm and pain, and to improve balance and co-ordination. An orthesis or back support may be used to correct kyphosis and decrease pain. Medication for pain is often needed and should cover both acute severe pain following fracture and chronic pain caused by postural change. A hip fracture is the most severe consequence of osteoporosis. The risk of hip fracture can be decreased by pharmacological treatment to increase bone mass and bone strength. However, in the very elderly the occurrence of falling may be more important than the failure of bone strength. Hip protectors have recently become available and have been shown to decrease the risk of hip fracture after a fall. These shunt the energy from the trochanter away to the sides. Non-pharmacological approaches to treatment are often neglected in daily practice, the emphasis being instead on treatment with drugs that decrease bone resorption and thereby increase bone strength. PMID- 11035907 TI - Glucocorticoid-induced osteoporosis. AB - Glucocorticoid drugs interact with bone metabolism at many levels, but their principal action is to reduce osteoblast number and bone matrix synthesis. Virtually all patients receiving glucocorticoids in doses above 5 mg per day lose bone, the amount lost being dependent on the cumulative steroid dose. The risk of fracture is also related to the individual's initial bone density, which in turn reflects race, sex, age, menopausal status, body weight, smoking and the nature of any underlying illness. Bone density measurement and personal fracture history are the best predictors of future fracture risk. Steroid-induced bone loss is reversible, so measures to minimize the systemic steroid dose or to withdraw these drugs altogether should be pursued no matter how long an individual has been using them. Increasing the calcium intake to 1.5 g per day, encouraging them to stop smoking and take more exercise, and treating any vitamin D deficiency are sensible measures in all patients. In those at high risk, bisphosphonates are the best documented interventions, although sex hormone replacement is also effective and can be used alone or in addition to bisphosphonates. PMID- 11035908 TI - Osteoporosis and osteoporotic fractures in men: a clinical perspective. AB - The lifetime risk of any fracture of the hip, spine or distal forearm in men aged 50 years has been estimated to be 13%, compared with 40% in women. Although the overall incidence of osteoporosis is less in men than in women, the disease still represents an important public health problem. In particular, hip fractures are associated with substantial mortality and morbidity, even more so than in women. In male patients presenting with osteoporotic fractures, major causes of skeletal fragility, such as hypogonadism, glucocorticoid excess, primary hyperparathyroidism and alcohol abuse, can often be identified. In as many as 50% of osteoporotic men, however, no aetiology can be found: these men suffer from a syndrome commonly referred to as idiopathic osteoporosis, which is presumably related to some type of osteoblast dysfunction. Recent evidence indicates that the loss of skeletal integrity in ageing men may be partially related to endocrine deficiencies, including vitamin D, androgen and/or oestrogen deficiency. While the consequences of vitamin D or oestrogen deficiency in women have been well established, the skeletal impact of these (partial) age-related deficiencies in men remains to be clarified. Osteoporosis in elderly men is a multifactorial disease, as it is in women. The prevention of osteoporosis should therefore focus not only on increasing the bone strength, but also on decreasing the risk of falls. However, the prevention and therapy of osteoporotic disorders in men are virtually unexplored. To date, the use of specific osteoporotic drugs in osteoporotic men is still based on reasonable but untested assumptions. PMID- 11035909 TI - Lower peak bone mass and its decline. AB - There is evidence that two-thirds of the risk of osteoporotic fracture can be predicted from the pre-menopausal bone mineral density. The frequency of osteoporosis in older women may be modifiable by implementing invention strategies in the pre- and peri-menopausal periods. Lower peak bone mineral density and bone loss can be identified in women with altered reproductive hormone or calciotrophic hormone concentrations, or selected lifestyle practices. Alterations in reproductive hormones may occur in adolescence (from an early age of pregnancy or the amenorrhoea of anorexia nervosa or exercise), in the pre menopause (nulliparity, oophorectomy, early ovarian failure or marginal hormonal status) or in the peri-menopause. Alterations in calciotrophic hormone concentrations include corticosteroid therapy and breast cancer treatment. Lifestyle risk factors include the misuse of alcohol and possibly smoking, physical inactivity or an imbalance in dietary intake. Effective intervention currently consists of treating underlying conditions and monitoring high-risk groups. PMID- 11035910 TI - Index PMID- 11035911 TI - Preface PMID- 11035912 TI - CeReS-18, a cell regulatory sialoglycopeptide, inhibits proliferation and migration of rat vascular smooth muscle cells. AB - CeReS-18, a cell regulatory sialoglycopeptide, has been shown to inhibit proliferation of a wide array of target cells. In the present study, the effect of CeReS-18 on vascular smooth muscle cell (SMC) proliferation was characterized in cultured rat aorta SMCs (A7r5). More extensively, the effect of CeReS-18 on platelet-derived growth factor (PDGF)-induced SMC migration was examined using a modified Boyden's chamber assay. CeReS-18 inhibits both SMC proliferation and migration in a concentration-dependent, calcium-sensitive, and reversible manner. Furthermore, cells preincubated with the inhibitor had an increased sensitivity to CeReS-18-mediated inhibition of SMC migration. Immunoprecipitation and in vitro phosphorylation assays demonstrated that MAP kinase activity was inhibited in the CeReS-18-treated cells and pretreatment with CeReS-18 suppressed the activation of MAP kinase stimulated by PDGF. However, it is not likely that the suppression of the MAP kinase pathway was directly responsible for the ability of CeReS-18 to inhibit migration of the rat aorta smooth muscle cells since a MEK specific inhibitor, PD98059, did not influence A7r5 cell migration. PMID- 11035913 TI - Disruption of cell-substrate adhesion activates the protein tyrosine kinase pp60(c-src). AB - Treatment of confluent chicken embryo fibroblasts (CEFs) with trypsin results in a dose- and time-dependent increase in c-Src protein tyrosine kinase (PTK) activity. A similar, but less marked, increase in c-Src PTK activity occurs upon incubation of CEFs in calcium-free phosphate-buffered saline, which also causes a decrease in cell-substrate adhesion. The increase in c-Src PTK activity following disruption of cell-substrate adhesion correlates with a decrease in the phosphorylation of c-Src at the regulatory site, Tyr527. The phosphotyrosine phosphatase inhibitor phenylarsine oxide blocks the increase in c-Src PTK activity seen following treatment with trypsin and the morphological changes associated with the disruption of cell-substrate adhesion. In contrast, disruption of cell-substrate adhesion causes a decrease in FAK PTK activity that rapidly returns to control levels when the cells are plated on fibronection coated dishes. Treatment of cells with cytochalasin D, which disrupts actin filaments but not cell-substrate adhesion, causes only a slight increase in c-Src PTK activity. Thus, these studies demonstrate a ligand-independent mechanism for the activation of c-Src that is consistent with its role in both cell adhesion and cell motility. Furthermore, these data suggest that similar to adhesion, loss of adhesion is not a passive process but can activate specific signaling pathways that may have significant effects on cellular function. PMID- 11035914 TI - The receptor-associated protein (RAP) interacts with several resident proteins of the endoplasmic reticulum including a glycoprotein related to actin. AB - The receptor-associated protein (RAP) is a chaperone found primarily in the endoplasmic reticulum (ER) that plays a necessary role in the folding and exocytic trafficking of members of the LDL receptor gene family including megalin and the LDL receptor-related protein (LRP). Recently, RAP has been shown to interact with a growing number of proteins including several that are unrelated to the LDL receptor family as well as new members of this rapidly expanding family. Based on these observations, we have applied chemical crosslinking procedures to identify additional novel RAP-interacting proteins, and thereby better characterize the scope of RAP's ER-related function. In this study, we have identified eight proteins with molecular weights of 32, 35, 46, 55, 70, 95, 170, and 200 kDa that interact with endogenous RAP. These proteins were found to associate with RAP in multiple cell types from different species, suggesting that their expression and interactions with RAP are ubiquitous. Results of pulse-chase experiments show that most of the proteins remain sensitive to endoglycosidase-H digestion, and also remain stably associated with RAP over an extended period, suggesting that they are ER resident proteins. All of the RAP-associated proteins appear to be largely soluble as they partition into the aqueous phase following TX-114 detergent extraction. Sequence analysis and immunoblotting of the 46-kDa RAP-associated glycoprotein (gp46) shows that it is structurally and immunologically related to actin. If gp46 is also functionally related to actin as an intracellular structural protein, it may represent a novel component of the putative ER matrix. PMID- 11035915 TI - Arrest of S-phase progression is impaired in Fanconi anemia cells. AB - Fanconi anemia (FA) is an inherited cancer-susceptibility disorder, characterized by genomic instability, hypersensitivity to DNA cross-linking agents, and a prolonged G2 phase of the cell cycle. We observed a marked dose-dependent accumulation of FA cells in the G2 compartment after treatment with 4,5',8 trimethylpsoralen (Me(3)Pso) in combination with 365 nm irradiation. Using bivariate DNA distribution methodology, we determined the proportion of replicating and arresting S-phase cells and observed that, whereas normal cells arrested DNA replication in the presence of Me(3)Pso cross-links and monoadducts, FA lymphoblasts failed to arrest DNA synthesis. Taken together, the above data suggest that, in response to damage induced by DNA cross-linking agents, the S phase checkpoint is inefficient in FA cells. This would lead to accumulation of secondary lesions, such as single- and double-strand breaks and gaps. The prolonged time in G2 phase seen in FA cells therefore exists in order to allow the cells to remove lesions which accumulated during the preceding abnormal S phase. PMID- 11035916 TI - Damage-resistant DNA synthesis in Fanconi anemia cells treated with a DNA cross linking agent. AB - Fanconi anemia (FA) is a recessive disorder associated with diverse congenital anomalies, progressive bone marrow failure, and a marked predisposition to develop cancer. At the cellular level, FA is characterized by a prolonged G(2) phase in proliferating cells and a marked hypersensitivity to both the cytotoxic and the clastogenic effects of agents which produce DNA interstrand cross-links. Treatment with these agents leads to even further prolongation of the G(2) phase in FA cells. We now show that FA cells, from four different complementation groups, fail to decrease their rates of replicative DNA synthesis, as do normal cells, following treatment with a DNA cross-linking agent. This may be responsible for the prolongation of the G2 phase seen in these cells, and suggests that the fundamental defect in response of FA cells to DNA cross-linking agents may be in the S phase, rather than the G(2) phase, of the cell cycle. PMID- 11035917 TI - HSP70 is involved in the control of chromosomal transcription in the amphibian oocyte. AB - The amphibian oocyte represents an excellent model for the study of transcription regulation. Indeed, any modification of transcriptional activity is directly reflected in lampbrush chromosome structure by concomitant morphological changes. Previous studies have led to the hypothesis of a putative role for heat-shock proteins HSP70 and/or HSC70 in transcriptional processes in the oocyte. In order to dissect out the relative role of HSP70 or HSC70 in these processes, we used an oligo-antisense strategy to specifically inhibit the function of the targeted protein. Effects of hsc70 and hsp70 antisense oligodeoxynucleotides were analyzed in terms of both mRNA quantity and protein synthesis. Their effects on oocyte transcription were analyzed at the level of structural organization of lampbrush chromosomes and nucleolar transcriptional activity. Our results show that specific inactivation of hsc70 mRNA by hsc70 antisense oligos led to a reversible inhibition of lampbrush chromosome transcription. However, such reversible inhibition of transcription is considered non-sequence specific since it is also induced by any oligo. In contrast, specific inactivation of hsp70 mRNA by hsp70 antisense oligos, which is correlated with a drop of HSP70 neosynthesis, results in an irreversible inhibition of lampbrush chromosome transcription. Furthermore, our results show that the inactivation of hsp70 or hsc70 mRNAs does not affect nucleolar transcription. Such data suggest a role for HSP70 in the control of chromatin modifications related to RNA polymerase II transcriptional activity. PMID- 11035918 TI - Heterotopic expression of the Xl-Fli transcription factor during Xenopus embryogenesis: modification of cell adhesion and engagement in the apoptotic pathway. AB - In the Xenopus laevis embryo, the overexpression of the Xl-FLI protein, a transcription factor of the ETS family, provokes severe developmental anomalies, which affect anteroposterior and dorsoventral polarities, optic cup formation, head cartilage morphogenesis, and erythrocyte differentiation. It has been proposed that these effects could be correlated to modifications of cell adhesion properties and/or to an increased engagement of cells in the apoptotic pathway during early development (Remy et al., Int. J. Dev. Biol. 40, 577-589, 1996). To address these questions, we have first analyzed the behavior of cells overexpressing the protein in both aggregation and adhesion assays. We observe perturbations of cell-cell interactions as well as perturbations of cell adhesion and spreading on fibronectin and extracellular matrix (ECM). Second, we have analyzed apoptosis of cells overexpressing the Xl-FLI protein, by testing DNA fragmentation, caspase-3 activity and by performing TUNEL assay. We show that Xl Fli overexpression results in the appearance of hallmarks of apoptosis, including exclusion of cells from the interior of the embryo, internucleosomal fragmentation of DNA and dose-dependent induction of caspase-3, resulting in the hydrolysis of poly(ADP-ribose) polymerase. In addition, a dominant-negative mutation of BMPs receptors decreases the effects of Xl-Fli overexpression, suggesting that a modification of the BMP signalling could be responsible for increased apoptosis. The latter appears to affect predominantly ventral and ventrolateral regions of the embryo. PMID- 11035919 TI - Segregation of RNA and separate packaging of DNA and RNA in apoptotic bodies during apoptosis. AB - Apoptosis is characterized by a complex and remarkably ordered choreography of events consisting of the preparatory and execution steps that all culminate in disposal of the cell remnants. The disposal occurs in a manner that is the least destructive to the tissue: the remains of nuclear chromatin and cytoplasm are packaged in apoptotic bodies which are then phagocytized by neighboring live cells without invoking inflammatory or autoimmune response. In the present study we describe that in the course of apoptosis cellular RNA becomes sequestered and packaged into granules and then into apoptotic bodies, separately from DNA. This separation, which appears to be initiated by the nucleolar segregation, was observed in HL-60 cells that were undergoing spontaneous apoptosis in cultures or were treated with the DNA-damaging drug, DNA topoisomerase I inhibitor camptothecin (CPT), or with the cell death ligand, tumor necrosis factor-alpha. RNA separation was also observed in apoptotic MCF-7 cells following treatment with CPT. RNA and DNA in apoptotic cells were identified histochemically, by their differential stainability with pyronin Y and Hoechst 33342 fluorochromes, respectively, and immunocytochemically, by labeling the RNA with BrU for various periods of time and detection of the incorporated precursor with fluoresceinated anti-BrU mAb; DNA was counterstained with 7-aminoactinomycin D. Over 90% of apoptotic bodies that contained RNA had no detectable DNA and vice versa, the apoptotic bodies containing DNA had no detectable RNA. Packaging RNA and DNA into separate apoptotic bodies suggests that the phagosomes of the cells that ingest these particles are specialized: some of them are responsible for DNA degradation, others for degradation of RNA. Such specialization may facilitate heterophagic degradation of nucleic acids during apoptosis. PMID- 11035920 TI - Pivotal role of the RB family proteins in in vitro thyroid cell transformation. AB - Rat thyroid differentiated cells (PC Cl 3) are an excellent model system with which to study the interaction between differentiation and cell transformation. We previously demonstrated that PC Cl 3 cells expressing the adenovirus E1A gene no longer depend on thyrotropin for growth and do not express thyroid differentiation markers. Here we show that an E1A mutant unable to bind the RB protein failed to transform the PC Cl 3 cells. Conversely, mutations in the E1A p300 interacting region did not affect its transforming ability. The pivotal role of RB family proteins in the thyroid cell transformation is supported by the thyrotropin independence induced by the E7 gene of human papilloma virus type 16, but not by a mutated form in the RB-binding region. PMID- 11035921 TI - Nitric oxide mediates laminin-induced neurite outgrowth in PC12 cells. AB - Laminin is a potent stimulator of neurite outgrowth in a variety of primary neurons and neuronal cell lines. Here, we investigate the role of nitric oxide in the signaling mechanism of laminin-mediated neurite outgrowth in the PC12 cell line. Within 8 s of exposure to laminin, PC12 cells produce nitric oxide. Peak laminin-induced nitric oxide levels reach 8 nM within 12 s of exposure to laminin and constitutive nitric oxide production is sustained for 1 min. A neurite outgrowth promoting synthetic peptide (AG73), derived from the laminin-1-alpha globular domain, also stimulated nitric oxide release. The nitric oxide synthase inhibitor, 1-NAME, prevents the formation of nitric oxide and here, 1-NAME inhibited both laminin-mediated and AG73-mediated neurite outgrowth by 88 and 95%, respectively. In contrast, C16, a synthetic peptide derived from the laminin 1-gamma chain, is shown here to promote PC12 cell attachment, but not neurite outgrowth. Interestingly, the C16 peptide did not activate nitric oxide release, suggesting that laminin-induced nitric oxide release in PC12 cells is associated only with neurite outgrowth promoting laminin domains and signals. In addition, the data here show that the nitric oxide released by PC12 cells in response to laminin is required as a part of the mechanism of laminin-mediated neurite outgrowth. PMID- 11035922 TI - Nuclear envelope fission is linked to cytokinesis in budding yeast. AB - We have investigated the relationship between nuclear envelope fission and cytokinesis during mitotic cell division in budding yeast. By carrying out time lapse and optical sectioning video microscopy analysis of cells that express green fluorescent protein (GFP)-tagged nuclear envelope and actomyosin ring components, we found that nuclear division is temporally coupled to cytokinesis. Light and electron microscopy analysis also showed that nuclear envelope fission and the division of the nucleoplasm are severely delayed in cytokinesis mutants, resulting in discoupling between the nuclear division cycle and the budding cycle. These results suggest that homotypic membrane fusion may be activated by components or the mechanical action of cytokinetic structures and presents a mechanism for the equal partitioning of the nucleus and the temporal coordination of this event with chromosome segregation during mitosis. PMID- 11035923 TI - Mitf is expressed in osteoclast progenitors in vitro. AB - Microphthalmia mutant (mi/mi) mice reveal defects in osteoclastogenesis and exhibit osteopetrosis. However, there have been no studies to test the importance of Mitf in in vitro osteoclastogenesis using the cells derived from mi/mi mice. Therefore, we investigated in vitro osteoclastogenesis using the cells derived from mi/mi mice. We cocultured spleen cells prepared from either wild-type or mi/mi mice with ST2 or TM8 stromal cells and found that formation of TRAP positive cells was significantly reduced in the cocultures of mi/mi spleen cells compared to wild-type spleen cells in the presence of 1,25(OH)(2) vitamin D(3) (vitamin D). We further investigated Mitf expression by Northern blot analysis in relation to the differentiation of osteoclasts using the cocultures of bone marrow cells with stromal/osteoblastic cells and found positive correlation in expression levels of c-fms and Mitf. Moreover, osteoclast-progenitor-like C7 cells expressed c-fms as well as Mitf mRNAs when cultured alone. C7 cells also expressed Mitf protein in their nuclei. Similar results were obtained when we used primary spleen cells, which differentiate into osteoclasts cultured in the presence of M-CSF and RANKL/ODF. Mitf expression levels in the cocultures of C7 cells and ST2 cells were not changed by treatment with vitamin D in the presence or absence of dexthamethasone. These results suggest that Mitf is expressed in osteoclast progenitors and its presence facilitates osteoclastogenesis. PMID- 11035925 TI - The tkNeo gene, but not the pgkPuro gene, can influence the ability of the beta globin LCR to enhance and confer position-independent expression onto the beta globin gene. AB - Whether drug-selectable genes can influence expression of the beta-globin gene linked to its LCR was assessed here. With the tkNeo gene placed in cis and used to select transfected cells, the beta-globin gene was expressed fourfold lower when it was positioned upstream of the LCR rather than downstream. This difference did not occur when the pgkPuro gene replaced tkNeo. Moreover, the beta globin gene situated upstream of the LCR was transcribed without position effects when it was cotransfected with a pgkPuro-containing plasmid, whereas cotransfection with a tkNeo plasmid gave measurable position effects. Previous results from transfected cells selected via a linked tkNeo gene suggested that the 3' end of the beta-globin gene has no impact on LCR-enhanced expression. Here, removal of the 3' end of the beta-globin gene resulted in lower and much more variable expression in both transgenic mice and cells cotransfected with pgkPuro. Together, the results suggest that tkNeo, but not pgkPuro, can strongly influence expression of the beta-globin gene linked to its LCR. The findings could partly explain why data on beta-globin gene regulation obtained from transfected cells have often not agreed with those obtained using transgenic mice. Hence, one must be careful in choosing a drug-selectable gene for cell transfection studies. PMID- 11035924 TI - Rac1-induced endocytosis is associated with intracellular proteolysis during migration through a three-dimensional matrix. AB - Transfection of Rat1 fibroblasts with an activated form of rac1 (V12rac1) stimulated cell migration in vitro compared to transfection of Rat1 fibroblasts with vector only or with dominant negative rac1 (N17rac1). To investigate the involvement of proteases in this migration, we used a novel confocal assay to evaluate the ability of the Rat1 transfectants to degrade a quenched fluorescent protein substrate (DQ-green bovine serum albumin) embedded in a three-dimensional gelatin matrix. Cleavage of the substrate results in fluorescence, thus enabling one to image extracellular and intracellular proteolysis by living cells. The Rat1 transfectants accumulated degraded substrate intracellularly. V12rac1 increased accumulation of the fluorescent product in vesicles that also labeled with the lysosomal marker LysoTracker. Treatment of the V12rac1-transfected cells with membrane-permeable inhibitors of lysosomal cysteine proteases and a membrane permeable selective inhibitor of the cysteine protease cathepsin B significantly reduced intracellular accumulation of degraded substrate, indicating that degradation occurred intracellularly. V12rac1 stimulated uptake of dextran 70 (a marker of macropinocytosis) and polystyrene beads (markers of phagocytosis) into vesicles that also labeled for cathepsin B. Thus, stimulation of the endocytic pathways of macropinocytosis and phagocytosis by activated Rac1 may be responsible for the increased internalization and subsequent degradation of extracellular proteins. PMID- 11035926 TI - Fine structural in situ analysis of nascent DNA movement following DNA replication. AB - Nascent DNA (newly replicated DNA) was visualized in situ with regard to the position of the previously replicated DNA and to chromatin structure. Localization of nascent DNA at the replication sites can be achieved through pulse labeling of cells with labeled DNA precursors during very short periods of time. We were able to label V79 Chinese Hamster cells for as shortly as 2 min with BrdU; Br-DNA, detected by immunoelectron microscopy, occurs at the periphery of dense chromatin, at individual dispersed chromatin fibers, and within dispersed chromatin areas. In these regions DNA polymerase alpha was also visualized. After a 5-min BrdU pulse, condensed chromatin also became labeled. When the pulse was followed by a chase, a larger number of gold particles occurred on condensed chromatin. Double-labeling experiments, consisting in first incubating cells with IdU for 20 min, chased for 10 min and then labeled for 5 min with CldU, reveal CldU-labeled nascent DNA on the periphery of condensed chromatin, while previously replicated IdU-labeled DNA has been internalized into condensed chromatin. Altogether, these results show that the sites of DNA replication correspond essentially to perichromatin regions and that the newly replicated DNA moves rapidly from replication sites toward the interior of condensed chromatin areas. PMID- 11035927 TI - Bone morphogenetic proteins secreted by breast cancer cells upregulate bone sialoprotein expression in preosteoblast cells. AB - It is well established that bone metastases comprise bone; however, the exact factors/mechanisms involved remain unknown. We hypothesized that tumor cells secreted factors capable of altering normal bone metabolism. The aims of the present study were to (1) determine the effects of secretory products isolated from HT-39 cells, a human breast cancer cell line, on osteoprogenitor cell (MC3T3 E1 cells) behavior, and (2) identify tumor-derived factor(s) that alters osteoblast activities. Conditioned media (CM) from HT-39 cells were collected following a 24-h serum-free culture. The ability of CM to alter gene expression in MC3T3-E1 cells was determined by Northern analysis. CM effects on cell proliferation and mineralization ability were determined using a Coulter counter and von Kossa stain, respectively. MC3T3-E1 cells were treated with CM plus noggin, a factor known to block bone morphogenic proteins (BMPs), to determine whether BMPs, shown to be present in CM, were linked with CM effects on MC3T3-E1 cell activity. In addition, inhibitors of MAP kinase kinase (MEK), protein kinase C (PKC), and protein kinase A were used to identify the intracellular signaling pathway(s) by which the active factors in CM regulated osteoblast behavior. CM treatment significantly enhanced BSP mRNA (2.5-fold over control), but had no effect on cell proliferation. Mineralization assay showed that CM enhanced mineral nodule formation compared to controls. Noggin inhibited CM-induced upregulation of BSP mRNA, suggesting that BMPs were responsible for upregulating BSP gene expression in MC3T3-E1 cells. The PKC inhibitor blocked CM-mediated upregulation of BSP, suggesting involvement of the PKC pathway in regulating BSP expression. BMPs secreted by HT-39 cells may be responsible for enhancing BSP expression in MC3T3-E1 cells. Continued studies targeted at determining the role of BMPs in regulating bone metabolism are important for understanding the pathogenesis of bone diseases. PMID- 11035928 TI - F9 embryonal carcinoma cells engineered for tamoxifen-dependent Cre-mediated site directed mutagenesis and doxycycline-inducible gene expression. AB - The study of gene functions in complex genetic environments such as mammalian cells would greatly benefit from systems allowing a tight control of gene expression. The tetracycline-inducible gene expression system and the site specific Cre/loxP recombination system have gained increasing popularity for conditional expression and gene disruption. To facilitate the analysis of gene functions in a cell autonomous system, we have established an F9 murine embryonal carcinoma cell line, constitutively expressing both the doxycycline-controlled transactivator rtTA and the tamoxifen-dependent Cre recombinase Cre-ER(T). The expression of a reporter gene placed under the control of tetracycline operators was induced about 1000-fold by doxycycline, and tamoxifen-induced excision of a loxP-flanked DNA segment occurred in all cells. This genetically engineered cell line, which allows, upon simple ligand addition, sophisticated genetic manipulations, such as sequential inactivation of loxP-flanked genes, and tightly controlled reexpression of their cDNAs, should be a valuable tool for studying mammalian gene functions. PMID- 11035929 TI - A novel assay to demonstrate an intersection of the exocytic and endocytic pathways at early endosomes. AB - The mechanism of transport of membrane proteins from the trans-Golgi to the cell surface is still poorly understood. Previous studies suggested that basolateral membrane proteins, such as the transferrin receptor and the asialoglycoprotein receptor H1, take an indirect route to the plasma membrane via an intracellular, most likely endosomal intermediate. To define this compartment we developed a biochemical assay based on the very definition of endosomes. The assay is based on internalizing anti-H1 antibodies via the endocytic cycle of the receptor itself. Internalized antibody formed immune complexes with newly synthesized H1, which had been pulse-labeled with [(35)S]sulfate and chased out of the trans Golgi for a period of time that was insufficient for H1 to reach the surface. Hence, antibody capture occurred intracellularly. Double-immunofluorescence labeling demonstrated that antibody-containing compartments also contained transferrin and thus corresponded to early and recycling endosomes. The results therefore demonstrate an intracellular intersection of the exocytic and endocytic pathways with implications for basolateral sorting. PMID- 11035930 TI - Downregulation of KRC induces proliferation, anchorage independence, and mitotic cell death in HeLa cells. AB - The large zinc finger protein KRC regulates transcription of target genes via the kappaB gene enhancer element. As an attempt to investigate the cellular function of KRC, we have established cell lines stably transfected with KRC expression vectors. Introduction of a vector directing expression of a transcript antisense to KRC mRNAs in several mammalian cell lines resulted in accelerated proliferation. Furthermore, in HeLa cells, downregulation of KRC conferred anchorage-independent growth and promoted cell cycle progression without an intervening cytokinesis, culminating in the formation of multinucleated giant cells. Ultimately these cells died. PMID- 11035931 TI - Loss of chromosome 13 in cultured human vascular endothelial cells. AB - In the vascular endothelium of human beings, telomere length is negatively related while the frequency of aneuploidy is positively related to donor age. Both in culture and in vivo the frequency of aneuploidy increases as telomere length is shortened. In this study we explored the relation between telomere length and aneuploidy in cultured human umbilical vein endothelial cells (HUVEC) by: (a) karyotype analysis and fluorescent in situ hybridization (FISH), (b) measurement of the terminal restriction fragments (TRF), and (c) assessment of replicative senescence by the expression of beta-galactosidase. Of 8 HUVEC strains, 7 cell strains lost chromosome 13, as shown by metaphase analysis and FISH of interphase cells. Five strains gained chromosome 11. In addition, five HUVEC strains became hypotetraploid shortly after the loss of chromosome 13. The loss of chromosome 13 was observed as early as PD 20, when mean TRF length was greater than 9 kb and the percentage of cells positive for beta-galactosidase was relatively low. The almost uniform loss of chromosome 13 suggests that this unique type of aneuploidy of HUVEC is the result of a progressive expression of clones with survival advantage. PMID- 11035932 TI - Cell adhesion molecule CEACAM1 associates with paxillin in granulocytes and epithelial and endothelial cells. AB - CEACAM1 functions as an epithelial tumor suppressor and as an angiogenic growth factor. In the present study, utilizing differentially (serine/threonine or tyrosine) phosphorylated cytoplasmic domains of CEACAM1 and CEACAM3 as bait to isolate associated proteins from granulocyte extracts, we have identified human paxillin as a binding partner of the tyrosine-phosphorylated cytoplasmic CEACAM1 domain. CEACAM1-paxillin complexes were coimmunoprecipitated from extracts of granulocytes, the colonic cell line HT29, and HUVECs. We identified phosphorylated Tyr-488-a residue in the cytoplasmic CEACAM1 domain known to be essential for the tumor suppressive effect-to be necessary for this association. The CEACAM1-paxillin interaction was confirmed using laser scanning confocal microscopy analyses in granulocytes and HT29 cells, where CEACAM1 colocalizes with paxillin at the plasma membrane. In HUVECs a highly polarized expression pattern and colocalization of paxillin and CEACAM1 was observed. These findings support the findings that CEACAM1 is linked to the actin-based cytoskeleton. PMID- 11035933 TI - Laminin-1 activates Cdc42 in the mechanism of laminin-1-mediated neurite outgrowth. AB - Here, we investigated the role of the small Rho GTPases Rac, Cdc42, and Rho in the mechanism of laminin-1-mediated neurite outgrowth in PC12 cells. PC12 cells were transfected with plasmids expressing wild-type and dominant-negative mutants of Rac (RacN17), Cdc42 (Cdc42N17), or Rho (RhoN19). Over 90% of the dominant negative Rho- and Rac-transfected cells extended neurites when plated on laminin 1; however, none of the PC12 cells transfected with the dominant-negative Cdc42 mutant extended neurites. In cells cotransfected with plasmids expressing c-Jun N terminal kinase and wild-type Cdc42, laminin-1 treatment stimulated detectable levels of c-Jun phosphorylation. Further, cotransfection with c-Jun N-terminal kinase and the dominant-negative Cdc42 mutant blocked laminin-1-mediated c-Jun phosphorylation. Transfection with either wild-type Rac or the dominant-negative Rac did not effect c-Jun phosphorylation. These data demonstrate that Cdc42 is activated by laminin-1 and that Cdc42 activation is required in the mechanism of laminin-1-mediated neurite outgrowth. PMID- 11035934 TI - Isolation and characterization of the notch ligand delta4. AB - Notch signaling plays a critical role in a variety of developmental programs. In vertebrates, the complexity of the process is underscored by the existence of multiple Notch receptors and multiple ligands, each of which displays a distinct expression profile. Furthermore, the ligands can be subdivided into two families, the Serrate/Jagged family and the Delta family. Here we present the isolation of a novel Notch ligand, Delta4. Expression analyses indicate that mouse Delta4 is highly expressed in the eye and lung during embryogenesis and in the heart, lung, liver, and kidney of the adult. Functionally, Delta4 is indistinguishable from Jagged1 in its abilities to inhibit myogenesis and to stimulate transcription through Notch1 and the DNA binding protein CSL. PMID- 11035935 TI - Cellular distribution and karyophilic properties of matrix, integrase, and Vpr proteins from the human and simian immunodeficiency viruses. AB - Infections by human and simian immunodeficiency viruses (HIV and SIV) are independent of host cell division since the preintegration complex (PIC), containing the viral DNA, is able to undergo active nuclear import after viral entry. In order to clarify the mechanisms responsible for nuclear import of the PIC, we have analyzed the subcellular distribution and the karyophilic properties of its viral components, matrix protein (MA), integrase (IN), Vpr, and Vpx. Although MA has been reported to contain a nuclear localization signal, the MA/GFP fusions are excluded from the nucleus and associated with cellular membranes. In contrast, both HIV-1 and SIV IN and Vpr localize in the nucleus of transfected cells. Interestingly, only Vpx from SIVsm virus accumulate in the nucleus while SIVsm Vpr is uniformly distributed throughout nucleus and cytoplasm. Coexpression of MA, Vpr, and IN does not induce any change in their respective intracellular localizations. Finally, we confirm the karyophilic properties of HIV-1 IN and Vpr using an in vitro nuclear import assay. These results indicate that the viral proteins IN and Vpr, which are strongly associated with the viral DNA within PIC, may participate in the nuclear import of the HIV PIC. PMID- 11035936 TI - MYCN-related suppression of functional CD44 expression enhances tumorigenic properties of human neuroblastoma cells. AB - Highly malignant neuroblastoma tumors with MYCN amplification have been shown to downregulate the expression of the CD44 adhesion receptor. We have previously shown that MYCN amplified neuroblastoma cell lines either lack CD44 expression or express a nonfunctional, nonhyaluronic acid-binding CD44 receptor. By analysis of cells with manipulated expression of either CD44 or MYCN, we demonstrate that transfection of cells with a CD44 full-length cDNA construct produced a functional receptor in single copy MYCN cells and a nonfunctional CD44 receptor in MYCN amplified cells, similar to the CD44 receptor expressed by cells with enforced MYCN. Analysis of the in vivo growth properties of the transfectants revealed that the restoration of a functional CD44 receptor in nonamplified cells resulted in the suppression of in vivo cell growth, therefore linking the MYCN related lack of hyaluronic acid-binding function of CD44 to the highly tumorigenic properties of a subset of neuroblastoma cells. PMID- 11035937 TI - Fungal role models: A bouquet of foes and friends. Fifth European conference on fungal genetics, Arachon, France, March 25-29, 2000. PMID- 11035938 TI - Horizontal gene transfer and the evolution of secondary metabolite gene clusters in fungi: an hypothesis. PMID- 11035939 TI - Genetics and molecular physiology of the yeast Kluyveromyces lactis. AB - With the recent development of powerful molecular genetic tools, Kluyveromyces lactis has become an excellent alternative yeast model organism for studying the relationships between genetics and physiology. In particular, comparative yeast research has been providing insights into the strikingly different physiological strategies that are reflected by dominance of respiration over fermentation in K. lactis versus Saccharomyces cerevisiae. Other than S. cerevisiae, whose physiology is exceptionally affected by the so-called glucose effect, K. lactis is adapted to aerobiosis and its respiratory system does not underlie glucose repression. As a consequence, K. lactis has been successfully established in biomass-directed industrial applications and large-scale expression of biotechnically relevant gene products. In addition, K. lactis maintains species specific phenomena such as the "DNA-killer system, " analyses of which are promising to extend our knowledge about microbial competition and the fundamentals of plasmid biology. PMID- 11035940 TI - Chimeric pheromone receptors in the basidiomycete Schizophyllum commune. AB - The pheromone receptor system of the basidiomycete Schizophyllum commune is capable of ligand discrimination to confer mating specificity. The pheromone receptors of the B alpha locus were investigated for ligand discrimination in a strategy of domain swapping experiments. Several altered phenotypes of chimeric receptors have been found. These include constitutive pheromone receptors which need no ligand for activation of the downstream cascade of events. In addition, receptors still dependent on ligand were identified that had altered pheromone activation profiles, including promiscuous receptors that are activated by pheromones of all nine specificities, including the former self. In addition, highly discriminative receptors were created which are activated by only two of the eight non-self-specificities. The chimeric receptors identify the last third of the receptor as the determinant for B alpha 1 specificity, whereas B alpha 2 specificity resides in noncontiguous domains covering the first and middle parts of the receptor molecule. PMID- 11035941 TI - Molecular and functional analyses of incompatibility genes at het-6 in a population of Neurospora crassa. AB - Two closely linked genes, un-24 and het-6, associated with the het-6 heterokaryon incompatibility functional haplotype were examined in 40 Neurospora crassa strains from a Louisiana sugarcane field. Partial diploid analyses were used to determine that half of the strains were functionally Oak Ridge (OR) and half were non-OR and indistinguishable from the standard Panama (PA) form. PCR-based markers were developed to identify polymorphisms within both un-24 and het-6. Two common forms of each gene occur based on these molecular markers. Rare forms of both un-24 and het-6 were identified as variants of the non-OR form by a DNA transformation assay. The heterokaryon incompatibility function of haplotypes, based on partial diploid analyses, was perfectly correlated with the PCR-based markers at both loci. This correlation indicates that the two loci are in severe linkage disequilibrium in this population sample and may act as an incompatibility gene complex. Southern hybridizations using OR- and PA-derived cloned probes from the region that spans un-24 and het-6 showed that the apparent absence of recombination in this approximately 25-kbp region is associated with low levels of overall sequence identity between the PA and OR forms. PMID- 11035942 TI - Molecular typing of Histoplasma capsulatum isolated from infected bats, captured in Mexico. AB - The present paper represents data on the genetic polymorphism of 13 Histoplasma capsulatum isolates recovered from infected bats randomly captured in the Mexican states of Morelos, Puebla, and Oaxaca. The polymorphic DNA patterns were analyzed by two-primer RAPD-PCR (random amplified polymorphic DNA-polymerase chain reaction) method. To amplify the fungal genome by PCR, the following primer arrangements were used: 5'-AACGCGCAAC-3' and 5'-AAGAGCCCGT-3'; 5'-AACGCGCAAC-3' and 5'-GTTTCCGCCC-3'; or 5'-AACGCGCAAC-3' and 5'-GCGATCCCCA-3'. A common polymorphic DNA pattern of H. capsulatum was revealed in different assays. This pattern is shared by 7 H. capsulatum isolates recovered from different specimens of nonmigratory bats (Artibeus hirsutus) captured in a cave in Morelos, by 5 isolates recovered from infected migratory bats (Leptonycteris nivalis) captured in Morelos and Puebla, and by 1 isolate from another migratory bat (L. curasoae) captured in Oaxaca. This polymorphic DNA pattern of H. capsulatum could represent fungal markers for the geographic areas studied, and considering its distribution in three different states of the Mexican Republic, the role of bats as responsible for H. capsulatum spreading in nature, in relation to their movements and migrations besides their shelter habits, is suggested. Analyses of DNA patterns of H. capsulatum isolated from infected bats, from clinical cases, and from blackbird excreta, have shown a major relatedness between bats and clinical isolates, in contrast to those isolates from bird excreta. PMID- 11035943 TI - Programmed ascospore death in the homothallic ascomycete Coniochaeta tetraspora. AB - Immature asci of Coniochaeta tetraspora originally contain eight uninucleate ascospores. Two ascospore pairs in each ascus survive and mature, and two die and degenerate. Arrangement of the two ascospore types in individual linear asci is what would be expected if death is controlled by a chromosomal gene segregating at the second meiotic division in about 50% of asci. Cultures originating from single homokaryotic ascospores or from single uninucleate conidia are self fertile, again producing eight-spored asci in which four spores disintegrate, generation after generation. These observations indicate that differentiation of two nuclear types occurs de novo in each sexual generation, that it involves alteration of a specific chromosome locus, and that the change occurs early in the sexual phase. One, and only one, of the two haploid nuclei entering each functional zygote must carry the altered element, which is segregated into two of the four meiotic products and is eliminated when ascospores that contain it disintegrate. Fusion of nuclei cannot be random-a recognition mechanism must exist. More study will be needed to determine whether the change that is responsible for ascospore death is genetic or epigenetic, whether it occurs just before the formation of each ascus or originates only once in the ascogonium prior to proliferation of ascogenous hyphae, and whether it reflects developmentally triggered alteration at a locus other than mating type or the activation of a silent mating-type gene that has pleiotropic effects. Similar considerations apply to species such as Sclerotinia trifoliorum and Chromocrea spinulosa, in which all ascospores survive but half the spores in each ascus are small and self-sterile. Unlike C. tetraspora, another four-spored species, Coniochaetidium savoryi, is pseudohomothallic, with ascus development resembling that of Podospora anserina. PMID- 11035944 TI - Production and characterization of recombinant guamerin, an elastase-specific inhibitor, in the methylotrophic yeast Pichia pastoris. AB - The elastase-specific inhibitor, guamerin, was expressed and secreted into a culture medium using the methylotrophic yeast Pichia pastoris, and the resulting recombinant guamerin was purified from the culture media using a two-step procedure composed of a hydrophobic interaction and reverse-phase chromatography. Up to 90 g/L of dry cell weight, the guamerin-producing recombinant P. pastoris was cultivated and guamerin was secreted into the culture medium at a level of 0.69 g/L. The recombinant guamerin was highly purified (>98%) with a recovery yield of 68%. Analyses of the purified guamerin revealed the same N-terminal amino acid sequence, amino acid composition, and molecular mass as found in the native leech protein. The recombinant guamerin exhibited the tight binding to porcine pancreatic elastase. Furthermore, the recombinant guamerin did not produce a humoral immune response in mice. PMID- 11035945 TI - Expression in Escherichia coli, folding in vitro, and characterization of the carbohydrate recognition domain of the natural killer cell receptor NKR-P1A. AB - NKR-P1A is a homodimeric type II transmembrane protein of the C-type lectin family found on natural killer (NK) cells and NK-like T cells and is an activator of cytotoxicity. Toward structure determination by NMR, the recombinant carbohydrate-recognition domain (CRD) of NKR-P1A has been expressed in high-yield in Escherichia coli and folded in vitro. The purified protein behaves as a monomer in size-exclusion chromatography and is bound by the conformation sensitive antibody, 3.2.3, indicating a folded structure. A polypeptide tag at the N-terminus is selectively cleaved from the CRD after limited trypsin digestion in further support of a compact folded structure. The disulfide bonds have been identified by peptide mapping and electrospray mass spectrometry. These are characteristic of a long form CRD. The 1D NMR spectrum of the unlabeled CRD and the 2D HSQC spectrum of the (15)N-labeled CRD are those of a folded protein. Chemical shifts of H(alpha) and NH protons indicate a considerable amount of beta strand structure. Successful folding in the absence of Ca(2+), coupled with the lack of chemical shift changes upon addition of Ca(2+), suggests that the NKR-P1A CRD may not be a Ca(2+)-binding protein. PMID- 11035946 TI - The delta-endotoxin proteins accumulate in Escherichia coli as a protein-DNA complex that can be dissociated by hydrophobic interaction chromatography. AB - The insecticidal protein CryIAc accumulated to form inclusion bodies in Escherichia coli upon overexpression of the cloned gene. The solubilized inclusion bodies contained the delta-endotoxin in association with DNA fragments of about 25 kb. The protein-DNA complex could be dissociated and the delta endotoxin purified by hydrophobic interaction chromatography on phenyl-Sepharose. The DNA was washed out in the high-salt buffer while the delta-endotoxin was bound to the matrix and was eluted at 4 degrees C by a stepwise decreasing potassium chloride gradient. The DNA-protein complex also contained plasmids harbored by the host strain. The plasmid DNA associated with the complex became competent to transform E. coli only after it was dissociated from the delta endotoxin. The hydrophobic interaction chromatography provides an efficient method for the purification of DNA-free activated toxin. PMID- 11035947 TI - Carica papaya glutamine cyclotransferase belongs to a novel plant enzyme subfamily: cloning and characterization of the recombinant enzyme. AB - A full-length cDNA encoding Carica papaya glutamine cyclotransferase was cloned by RT-PCR on the basis of results from amino acid sequencing of tryptic fragments of the native enzyme. The cDNA of 1036 nucleotides encodes a typical 22-residue signal peptide and a mature protein of 266 residues with a calculated molecular mass of 30,923 Da. Five plant ESTs encoding putative QCs highly homologous to PQC were identified and the numbers and locations of cysteines and N-glycosylation sites are conserved. The plant QC amino acid sequences are very different from the known mammalian QC sequences and no clear homology was observed. The PQC cDNA was expressed in Escherichia coli as either His-tagged PQC, with three different signal peptides and in fusions with thioredoxin, glutathione S-transferase, and (pre-) maltose-binding protein. In all cases, the expressed protein was either undetectable or insoluble. Expression in Pichia pastoris of PQC fused to the alpha-factor leader resulted in low levels of PQC activity. Extracellular expression of PQC in the insect cell/baculovirus system was successful and 15-50 mg/liter of active PQCs with three different secretion signals was expressed and purified. Further, PQC N-terminally fused to a combined secretion signal/His-tag peptide was correctly processed by the host signal peptidase and the His-tag could subsequently be removed with dipeptidyl peptidase I. The expressed products were characterized by activity assays, SDS-PAGE, N-terminal amino acid sequencing, MALDI-TOF mass spectroscopy, and peptide mass fingerprint analysis. PMID- 11035948 TI - Expression of functional soluble human alpha-globin chains of hemoglobin in bacteria. AB - Individual, soluble human alpha-globin chains were expressed in bacteria with exogenous heme and methionine aminopeptidase. The yields of soluble alpha chains in bacteria were comparable to those of recombinant non-alpha chains expressed under the same conditions. Molecular mass and gel-filtration properties of purified recombinant alpha chains were the same as those of authentic human alpha chains. Biochemical and biophysical properties of isolated alpha chains were identical to those of native human alpha chains as assessed by UV/vis, circular dichroism (CD), and nuclear magnetic resonance (NMR) spectroscopy which contrasts with previous results of refolded precipitated alpha chains made in the presence of heme in vitro (M. T. Sanna et al., J. Biol. Chem. 272, 3478-3486, 1997). Mixtures of purified, soluble recombinant alpha-globin and native beta-globin chains formed heterotetramers in vitro, and oxygen- and CO-binding properties as well as the heme environment of the assembled tetramers were experimentally indistinguishable from those of native human Hb A. UV/vis, CD, and NMR spectra of assembled Hb A were also the same as those of human Hb A. These results indicate that individual expressed alpha chains are stable in bacteria and fold properly in vivo and that they then can assemble with free beta chains to form hemoglobin heterotetramers in vivo as well as in vitro. PMID- 11035949 TI - Separation of copurifying GroEL from glutathione-S-transferase fusion proteins. AB - The purification of overexpressed fusion proteins using bacterial expression systems is a useful tool for the study of many proteins. One problem that can occur is the formation of stable interactions between the expressed fusion protein and certain endogenous bacterial proteins, such as the molecular chaperone GroEL. Such interactions may result in the copurification of contaminating bacterial proteins. Here we describe an efficient and inexpensive method for the removal of contaminating GroEL from a bacterially expressed GST fusion protein. In this method, denatured bacterial proteins are added to the bacterial lysates prior to the addition of glutathione Sepharose resin. The denatured proteins compete for GroEL binding, thereby releasing the GroEL contaminants from the expressed fusion protein. PMID- 11035950 TI - Expression, purification, and characterization of the human receptor activator of NF-kappaB ligand (RANKL) extracellular domain. AB - Receptor activator of NF-kappaB ligand (RANKL) is a type II transmembrane protein found on osteoblasts which functions as a major determinant of osteoclast differentiation and activation. RANKL mediates bone homeostasis through binding to the cognate ligand on osteoclasts, RANK, and a soluble decoy receptor, osteoprotegerin (OPG). We designed a construct encoding the extracellular domain of human RANKL that conformed to reports of native processing. To encourage folding and posttranslational modification of a normally membrane-inserted moiety, we expressed the RANKL truncate as a secreted protein using the signal sequence from OPG in a Trichoplusia ni cell line using a baculovirus expression vector. RANKL was purified by a three-step process including an OPG-Fc affinity column. SDS-PAGE and mass spectral analysis indicated that the protein was >99% pure and glycosylated. Circular dichroism spectra revealed that the protein exhibited structural elements similar to tumor necrosis factor-alpha. By BIAcore analysis, RANKL bound to OPG with an affinity of 6.7 nM. Sedimentation equilibrium analytical ultracentrifugation analyses established that our protein existed as a trimer. We conclude that our expressed human RANKL truncate is folded, is functional, and exhibits self-association consistent with other family members. PMID- 11035951 TI - Optimized expression and purification of toluene 4-monooxygenase hydroxylase. AB - Toluene 4-monooxygenase is a four-protein complex that catalyzes the O(2)- and NADH-dependent oxidation of toluene to p-cresol. The influence of various expression systems on the host cell growth characteristics, purified protein yields, and specific activity of the hydroxylase (T4moH) component of the complex was evaluated by considering the cell mass obtained per liter of fermentation culture medium, the purified protein obtained per gram of cell mass, and the specific activity of purified T4moH. The specific activity of purified T4moH was determined to be 1200-1250 nmol of p-cresol formed per minute per milligram of T4moH in air-saturated 50 mM phosphate buffer, pH 7.5, at 25 degrees C in the presence of optimal concentrations of the other protein components of the complex, saturating toluene (5.8 mM at 25 degrees C), and saturating NADH (1 mM). This value was obtained for T4moH purified from several different expression systems and apparently represents the maximal specific activity of the enzyme complex for toluene hydroxylation. By manipulation of vectors and gene inserts to eliminate adventitious catalytic turnover of NADH, up to 60-fold increase in the volumetric yield of T4moH activity was obtained from recombinant fermentations in Escherichia coli BL21(DE3). PMID- 11035952 TI - Purification of myristoylated and nonmyristoylated neuronal calcium sensor-1 using single-step hydrophobic interaction chromatography. AB - Neuronal calcium sensors (NCSs) belong to a family of Ca(2+)-binding proteins, which serve important functions in neurotransmission, and are highly conserved from yeast to humans. Overexpression of the neuronal calcium sensor-1, called frequenin in the fruit fly and in frog, increases the release of neurotransmitters. Studying the functional role of frequenin in mammals and understanding its structural dynamics is critically dependent on the availability of active purified protein. Neuronal calcium sensors like other members of the family share common structural features: they contain four EF-hands as potential binding sites for Ca(2+) and an N-terminal consensus sequence for myristoylation. Previously, recoverin, distantly related to NCSs, has been expressed and purified from Escherichia coli, involving a combination of different chromatographic steps. NCS-1 has earlier been purified adopting a two-step procedure used for recoverin purification. We have overexpressed NCS-1 from rat in its myristoylated and nonmyristoylated form in E. coli and purified it from crude lysates using a single-step hydrophobic interaction chromatography. The purified protein was identified by Western blotting and mass spectrometry and assayed for its ability to bind Ca(2+) using a Ca(2+) shift assay, terbium fluorescence, and Stains-all binding. The present protocol provides a rapid, more efficient and simplified, single-step method for purifying NCS-1 for structural and functional studies. This method can also be applied to purify related proteins of the superfamily. PMID- 11035953 TI - Roles of NADPH-P450 reductase in the O-deethylation of 7-ethoxycoumarin by recombinant human cytochrome P450 1B1 variants in Escherichia coli. AB - Four human cytochrome P450 1B1 (CYP1B1) allelic variants were purified from membranes of Escherichia coli in which respective CYP1B1 cDNAs and human NADPH P450 reductase cDNA have been introduced. Purified CYP1B1 variants were used to reconstitute 7-ethoxycoumarin O-deethylation activities with purified rabbit liver or recombinant (rat) NADPH-P450 reductase in the phospholipid vesicles and compared with those catalyzed by CYP1B1 enzymes in the membranes of E. coli in monocistronic (by adding the reductase) and bicistronic (without addition of extra reductase) systems. In the bicistronic system, the ratio of expression of NADPH-P450 reductase to CYP1B1 proteins was found to range from 0.2 to 0.5. Purified CYP1B1 enzymes (under optimal reconstitution conditions) catalyzed 7 ethoxycoumarin O-deethylation at rates one-third to one-fourth of those catalyzed by membranes of E. coli coexpressing CYP1B1 and the reductase proteins. Full catalytic activities in reconstituted systems were achieved with a twofold molar excess of NADPH-P450 reductase to CYP1B1; in membranes of E. coli with the monocistronic CYP1B1 construct, an eightfold molar excess of reductase to CYP1B1 was required. However, in membranes of bicistronic constructs, there was no additional stimulation of 7-ethoxycoumarin O-deethylation by extra NADPH-P450 reductase, despite the fact that the molar ratio of expression levels of reductase to CYP1B1 was <0.5. These results suggest that NADPH-P450 reductase produced in the bacterial membranes is more active in interacting with CYP1B1 proteins in the bicistronic system than the reductase added to artificial phospholipid vesicles or bacterial membranes. PMID- 11035954 TI - Recombinant maize protoporphyrinogen IX oxidase expressed in Escherichia coli forms complexes with GroEL and DnaK chaperones. AB - The clone corresponding to maize plastidic protoporphyrinogen IX oxidase (PPO) has been isolated by functional complementation and inserted into a pET16b vector for expression in Escherichia coli. Recombinant PPO was purified by standard affinity chromatography using a metal chelating resin. Two contaminants copurified with recombinant PPO and were identified as GroEL and DnaK. Since chaperone binding to hydrophobic regions of the protein is regulated by ATP availability, an ATP washing step was introduced prior to elution of the recombinant protein from an affinity column. This washing step selectively removed both chaperones and allowed the recovery of pure PPO. Coexpression of PPO and GroELS resulted in a sixfold increase of soluble PPO yield, suggesting that bacterial chaperones could be limiting during the folding of the heterologous protein. However, a portion of PPO was still found in the insoluble fraction. Buffer containing the GroEL and DnaK enabled resuspension of PPO from the insoluble fraction but failed to enhance refolding of the denaturated protein. Attempts to increase the amount of soluble PPO using a thioredoxin-PPO fusion protein were not successful. Initial characterization of the recombinant PPO found that it possessed a high V(max), an elevated affinity for substrate, and an elevated sensitivity to PPO inhibitor herbicides compared to previous reports. PMID- 11035955 TI - Purification and characterization of a luxO promoter binding protein LuxT from Vibrio harveyi. AB - Bioluminescence in the marine bacterium Vibrio harveyi is cell density dependent and is regulated by small molecules (autoinducers) excreted by the bacteria. The autoinducer signals are relayed to a central regulator, LuxO, which acts in its phosphorylated form as a repressor of the lux operon at the early stages of cell growth. We report in these studies the purification to homogeneity of a luxO DNA binding protein (LuxT) from V. harveyi after five major chromatography steps, including a highly effective DNA affinity chromatography step and reverse-phase HPLC. Regeneration of binding activity was accomplished after HPLC and SDS-PAGE by renaturation of LuxT from guanidine hydrochloride. It was also demonstrated that the functional LuxT was a dimer of 17 kDa that bound tightly (K(d) = 2 nM) to the luxO promoter. The sequences of three tryptic peptides obtained on digestion of the purified protein did not match any sequences in the Protein Data Bank, indicating that LuxT is a new V. harveyi lux regulatory protein. PMID- 11035956 TI - A mutant sarcosine oxidase in which activity depends on flavin adenine dinucleotide. AB - The covalent flavin attachment site in the Arthrobacter sarcosine oxidase (cysteine at position 318) was replaced with serine, and the mutational effect of C318S was analyzed. Wild type and C318S with a C-terminal 6-histidine tag were constructed and homogeneously purified by the single step. The covalently binding to flavin was not essential to the enzyme activity because the C318S mutant exhibited extremely weak activity. Moreover, the activity of the mutant was recovered in the presence of flavin adenine dinucleotide (FAD), and significantly increased as the concentration of FAD increased. This dependence of the mutant on FAD indicates that the noncovalent binding of free FAD to the mutant enzyme is reversible. PMID- 11035957 TI - Development of a recombinant bacterial expression system for the active form of a human transforming growth factor beta type II receptor ligand binding domain. AB - Expression systems have been designed to test the suitability of expressing the high cysteine containing extracellular domain (residues 1-136) of human transforming growth factor beta type II receptor (TbetaRII). Receptor expressed using a baculovirus system was functional following both enzymatic deglycosylation and elimination of the N-terminal 22 amino acids by protease degradation. Bacterial expression of a TbetaRII lacking the 26 N-terminal amino acids retained the ability to bind its ligand, TGF-beta1. Receptor expressed in bacteria was sensitive to proteolytic degradation at residue Lys98 but a K98T mutation eliminated degradation and did not disrupt binding. Although several different forms of TbetaRII were expressed, only a fusion with glutathione S transferase gave soluble TbetaRII, which was purified at a yield of 0.1 mg/10 L of bacterial growth. N-Terminal truncations of TbetaRII (residues 22-136 or 27 136) could be refolded from inclusion bodies and purified to an active form with an efficiency of 10%. PMID- 11035958 TI - Expression of recombinant galactose oxidase by Pichia pastoris. AB - Galactose oxidase catalyzes the oxidation of a variety of primary alcohols, producing hydrogen peroxide as a product. Among hexose sugars, the enzyme exhibits a high degree of specificity for the C6-hydroxyl of galactose and its derivatives, underlying a number of important bioanalytical applications. Galactose oxidase cDNA has been cloned for expression in Pichia pastoris both as the full-length native sequence and as a fusion with the glucoamylase signal peptide. Expression of the full-length native sequence results in a mixture of partly processed and mature galactose oxidase. In contrast, the fusion construct directs efficient secretion of correctly processed galactose oxidase in high density, methanol-induced fermentation. Culture conditions (including induction temperature and pH) have been optimized to improve the quality and yield (500 mg/L) of recombinant enzyme. Lowering the temperature from 30 to 25 degrees C during the methanol induction phase results in a fourfold increase in yield. A simple two-step purification and one-step activation produce highly active galactose oxidase suitable for a wide range of biomedical and bioanalytical applications. PMID- 11035959 TI - Expression and purification of histidine-tagged proteins from the gram-positive Streptococcus gordonii SPEX system. AB - Streptococcus gordonii (S. gordonii) has been used as a gram-positive bacterial expression vector for secreted or surface-anchored recombinant proteins. Fusion of the gram-positive bacterial N-terminal signal sequence to the target protein is all that is required for efficient export. This system is termed SPEX for Surface Protein EXpression and has been used to express proteins for a variety of uses. In this study, the SPEX system has been further developed by the construction of vectors that express polyhistidine-tagged fusion proteins. SPEX vectors were constructed with an N-terminal or C-terminal histidine tag. The C repeat region (CRR) from Streptococcus pyogenes M6 protein and the Staphylococcus aureus nuclease A (NucA) enzyme were tested for expression. The fusion proteins were purified using metal affinity chromatography (MAC). Results show that the fusion proteins were expressed and secreted from S. gordonii with the His tag at either the N- or C-terminal position and could be purified using MAC. The M6 fusions retained immunoreactivity after expression and purification as determined by immunoblots and ELISA analyses. In addition, NucA fusions retained functional activity after MAC purification. The M6-His and NucA-His fusions were purified approximately 15- and 10-fold respectively with approximately 30% recovery of protein using MAC. This study shows that the polyhistidine tag in either the N- or C-terminal position is a viable way to purify secreted heterologous proteins from the supernatant of recombinant S. gordonii cultures. This study further illustrates the value of the SPEX system for secreted expression and purification of proteins. PMID- 11035960 TI - Functional recombinant rabbit muscle phosphoglucomutase from Escherichia coli. AB - The gene coding for phosphoglucomutase (PGM) from Oryctolagus cuniculus (rabbit) has been expressed in Escherichia coli under a T7 expression system with a His tag. About half of the expressed PGM protein was present in inclusion bodies, but this protein was inactive when solubilized. The protein in the soluble cell fraction was isolated and purified in one step on a Ni-NTA column. The eluate from this column was adjusted to 95% saturated ammonium sulfate, and the resulting protein precipitate was resuspended in sodium phosphate buffer and dialyzed against 2.5 M ammonium sulfate. The final yield of protein was about 10 mg per liter of LB medium. The protein was judged to be greater than 90% pure on the basis of gel electrophoresis and activity measurements (128 U per milligram). Our motivation for developing this bacterial production system for PGM has been to prepare sufficient quantities of stable-isotope-labeled protein for experiments that utilize recently developed NMR technologies suitable for proteins the size of PGM (61.6 kDa). Preliminary NMR studies indicate that the current level of purity is adequate for this work. The construct described here was designed to incorporate an N-terminal His-tag for ease of isolation. Although PGM is a metalloprotein, the His-tag does not appear to interfere with activity. The presence of the His-tag should not pose a problem for proposed (31)P NMR investigations of the protein and its complexes in aqueous solution or incorporated into reverse micelles. However, we plan to design a cleavable His tag for later (1)H, (13)C, (15)N studies of the active site, which includes essential histidine residues. PMID- 11035961 TI - One-step purification of the recombinant catalytic subunit of pyruvate dehydrogenase phosphatase. AB - A facile one-step affinity chromatographic purification of the recombinant catalytic subunit (PDPc) of bovine pyruvate dehydrogenase phosphatase (PDP) to near homogeneity is described. PDPc binds in the presence of Ca(2+) to the inner lipoyl domain (L2) of the dihydrolipoamide acetyltransferase component (E2) of the mammalian pyruvate dehydrogenase complex. The affinity column consists of a glutathione S-transferase (GST)-L2 fusion protein bound to glutathione-Sepharose 4B beads. An extract of transformed Escherichia coli cells containing 50 mM Tris buffer (pH 7.5), 2 mM CaCl(2), 5 mM MgCl(2,) 150 mM NaCl, 0.5 mM dithiothreitol, 1% Triton X-100, and l M urea was passed through the affinity column, and the column was washed extensively with this buffer mixture. PDPc was eluted with 50 mM Tris buffer (pH 7.5) containing 5 mM MgCl(2), 0.5 mM dithiothreitol, and 1 mM EGTA. Approximately 22 mg of highly purified PDPc was obtained from 10 g (wet weight) of transformed cells. The preparation contained a small amount of a "nicked" form of PDPc. The cleavage is between Arg-394 and Arg-395. PMID- 11035962 TI - Intraoperative quality control of carotid endarterectomy. PMID- 11035963 TI - Cochrane collaborative review group on peripheral vascular disease: review abstracts. PMID- 11035964 TI - Is there a positive volume-outcome relationship in peripheral vascular surgery? Results of a systematic review. AB - OBJECTIVES: to examine the evidence for the existence, or otherwise, of a positive volume-outcome relationship in the area of peripheral vascular surgery. DESIGN: systematic overview of prospective or retrospective volume-outcome studies. DATA SOURCES: seven bibliographic databases were searched for English language articles published between 1986 and 1998. STUDY SELECTION: thirty-six articles published in peer-reviewed journals; excluding editorials, letters or abstracts; and addressing volume and outcome in peripheral vascular surgery. Criteria were applied and agreed by consensus between two of the authors. DATA EXTRACTION: the articles identified were independently assessed by two of the authors. Studies were categorised into three distinct areas - carotid endarterectomy- (17 studies), abdominal aortic aneurysm repair (16 studies) and other vascular interventions (four studies). Within each category studies were further classified according to full adjustment, partial adjustment or no adjustment for case mix. Where discrepancies arose, decisions were referred to a third author for arbitration. DATA SYNTHESIS: findings for carotid endarterectomy identified a positive volume-outcome relationship for both mortality and stroke at the physician level. There was less support for a positive relationship for mortality at the hospital level, and no evidence of benefits for stroke in higher volume hospitals. If only studies making a full adjustment for case mix are included, there is no clear support from statistically significant evidence for or against a positive volume-outcome relationship. For repair of unruptured abdominal aortic aneurysms there is evidence of a positive volume-outcome relationship at both the physician and hospital level, with evidence being particularly strong at the level of the hospital. For ruptured aneurysms the evidence is suggestive of there not being a positive volume-outcome relationship at the hospital level, while for physicians the evidence is more balanced with no clear support either way. For other vascular interventions there were insufficient studies (n=4) from which to draw meaningful conclusions. CONCLUSIONS: our results show that evidence of a relationship between volume and outcome in peripheral vascular surgery may be attributable to factors such as lack of adjustment for case-mix, different definitions of volume and poor quality of studies, especially those of retrospective design. Future studies should address these deficiencies by making full adjustment for case mix and by being prospective in design. PMID- 11035965 TI - Assessment of generic health-related quality of life in patients with intermittent claudication. AB - OBJECTIVES: to measure quality of life in patients with intermittent claudication and evaluate the ability of patients and vascular surgeons to make a similar assessment. DESIGN, MATERIALS AND METHODS: in this prospective study patients with intermittent claudication attending two vascular clinics were asked to complete a generic health-related quality of life instrument (MOS SF-36). Patient quality of life and vascular surgeons' assessment of patient quality of life were further evaluated using a single question/adjectival scale response combination. RESULTS: patients' self-assessment of their quality of life correlated better with the SF-36 score than did the surgeons' assessment. There was little correlation between the surgeons' and patients' own assessment of quality of life. The surgeons differed significantly from each other in their assessments. Claudicants had lower SF-36 scores than population norms in pain and physical aspects of quality of life. CONCLUSIONS: claudicants have worse quality of life than the general population, with pain and physical limitations being the most important domains. Surgeons predict the quality of life of claudicating patients less accurately than patients do themselves, and may differ from their colleagues in such assessments. Objective quality of life assessment in claudicants should be undertaken before treatment is decided. PMID- 11035966 TI - Optimal endothelialisation of a new compliant poly(carbonate-urea)urethane vascular graft with effect of physiological shear stress. AB - OBJECTIVE: to define the optimal seeding conditions of a new stress free poly(carbonate-urea)urethane (CPU) graft with compliance similar to that of human artery with honeycomb structure engineered during the manufacturing process to enhance adhesion and growth of endothelial cells. METHODS: (111)Indium-oxine radiolabeled human umbilical vein endothelial cells (HUVEC) were seeded onto CPU grafts at (a) concentrations from 2-24x10(5)cells/cm(2)and (b) incubated for 0.5, 1, 2, 4 and 6 h. Following incubation, graft segments were subjected to three washing/gamma counting procedures and scanning electron microscopy (SEM). Cell viability was measured using a modified Alamar blue(TM)assay. To test physiological retention a pulsatile flow phantom was used to subject optimally seeded (16x10(5), 4 h) CPU grafts to arterial shear stress for 6 h with real time acquisition of scintigraphic images of seeded grafts using a nuclear medicine gamma camera system. RESULTS: the seeding efficiency of 54+/-13% post three washes was achieved using 16x10(5)cells/cm(2). Similarly in SEM micrographs a seeding density of 16x10(5)cells/cm(2)resulted in a confluent monolayer. Seeded CPU segments incubated for 4 h exhibited significantly higher resistance to wash off than segments incubated for 30 min (p <0.05). Exposure of seeded grafts to pulsatile shear stress resulted in some cell loss with 67+/-3% of cells adherent following 6 h of perfusion with ongoing metabolic activity. Thus, optimal conditions were 16x10(5)cells/cm(2)at 4 h. CONCLUSIONS: the optimal seeding conditions have been defined for "tissue-engineered" vascular graft which allow complete endothelialisation and high cell-to-substrate strength that resists hydrodynamic stress. PMID- 11035967 TI - Occlusive arterial disease in HIV-infected patients: a preliminary report. AB - OBJECTIVES: to preliminarily describe the clinical features and management of arterial occlusive disease in human immunodeficiency virus (HIV) infected patients. MATERIALS: twenty HIV positive patients with symptomatic large-vessel arterial occlusion treated by a tertiary vascular unit in a 3-year period. METHODS: retrospective review of clinical case records. RESULTS: patients were noted to be young (median age 37 years), with preponderance of males. Twelve patients had evidence of advanced HIV infection. All patients had critical ischaemia, involving the upper limbs in four and the lower limbs in 16. Coagulation abnormalities were noted in two cases. Operative intervention in 18 patients included revascularisation in seven. Thrombotic occlusion of normal looking arteries was noted. Arterial biopsy revealed leucoIcytoclastic vasculitis indicative of HIV arteritis in three of five cases examined. CONCLUSIONS: initial experience with large-vessel occlusive disease in HIV positive patients suggests an underlying arteritic aetiology, with clinical and pathological features distinct from atherosclerosis. Further in-depth study is necessary to clarify the pathophysiological basis thereof. PMID- 11035968 TI - Surgical correction of isolated superficial venous reflux reduces long-term recurrence rate in chronic venous leg ulcers. AB - OBJECTIVES: surgical correction of isolated superficial venous reflux in ulcerated legs may reduce short term recurrence rates but the longer term benefits are unknown. DESIGN: prospective non-randomised cohort study. METHODS: consecutive patients with chronic leg ulcers were prospectively assessed at a one stop clinic over a 4-year period from July 1995 to July 1999. All patients with ankle brachial pressure indices (ABPI)50.85 were initially treated with weekly four-layer bandaging. Venous duplex studies in all ulcerated legs assessed venous reflux pattern with surgery being offered to all those with isolated superficial reflux, of whom 56% accepted. Patients were advised to wear class two elastic compression stockings after healing. RESULTS: 766 legs in 669 patients were assessed. Six hundred and thirty-three legs had an ABPI50.85, 236 (39%) demonstrating isolated superficial venous reflux. Surgery was performed on 131 of these legs. Twelve and 24 week healing rates were 50% and 72% for operated legs and 62% and 74% for non-operated legs (p=0.67; Kaplan-Meier life table analysis). Recurrence rates at 1, 2 and 3 years were 14%, 20% and 26% for operated legs and 28%, 30% and 44% for non-operated legs (p=0.03; Kaplan-Meier life table analysis). CONCLUSION: surgical correction of superficial venous reflux in legs with chronic leg ulceration may reduce ulcer recurrence rate at 1, 2 and 3 years. PMID- 11035969 TI - Optimal interval screening and surveillance of abdominal aortic aneurysms. AB - OBJECTIVES: to determine safe and optimal intervals of rescreening and surveillance for AAA. METHODS: hospital-based mass screening of 6339 65-73-year old men from 1994-98. 76.4% attended. One hundred and ninety-one (4%) had AAA53 cm. Twenty-four (0.5%) were initially >5 cm and referred for surgery, while the rest were offered annual control scans to check for expansion. Later, all 348 (7.5%) men who 3 to 5 years ago had an ectatic aorta (infrarenal aortic diameter of 25-29 mm or distal/renal aortic diameter ratio >1.2) were offered rescreening. Of these, 62 (18%) died before rescanning, while 248 of the survivors attended rescreening (87%). Furthermore, a random sample of 380 of those with non-ectatic aortas were offered rescreening. Of these, 49 (13%) died before rescreening (p=0.06), while 275 (83%) of the survivors attended re-screening. RESULTS: none of the controls had developed AAA. Of those who initially had an 25-29 mm aorta, 29% had developed AAA (size range 30-48 mm) with expansion rates varying from 1.0 to 4.7 mm/year. Only 3.5% with a ratio >1.2 developed AAA (size range: 30-34 mm) with expansion rates from 1.3 to 2.4 mm/year. During the fourth year of surveillance some AAA initially sized below 3.5 cm expanded to above 5 cm, while some sized 3.5-3.9 cm did so during the second year, >4 cm did so during the first year of surveillance. CONCLUSION: rescreening for AAA can be restricted to initially ectatic aortas sized 25-29 mm at 5-year intervals. Surveillance of small AAA can be restricted to 1-4 year intervals. PMID- 11035970 TI - A comparison of the mortality rate after elective repair of aortic aneurysms detected either by screening or incidentally. AB - OBJECTIVE: to compare predicted and actual mortality rates, using POSSUM scoring, after elective repair of abdominal aortic aneurysms (AAAs) detected from the Gloucestershire Aneurysm Screening Programme and those discovered incidentally. METHODS: a sample of 276 men undergoing elective AAA repair in Gloucestershire between 1991 and 1998 was studied. AAAs were either detected from the screening programme or were discovered incidentally and referred from other sources. Mortality data relating to these patients have been recorded prospectively. POSSUM scoring was performed retrospectively from patients>> notes in both groups and related to outcome (30 day and in-hospital mortality). POSSUM and P-POSSUM methodology were used to compare observed and predicted mortality rates. RESULTS: in the 276 men who had elective AAA repair, the overall mortality rate was 7%. Mortality was lower in screen-detected AAAs (3/111, 3%) than AAAs discovered incidentally (16/175, 9%) (p=0.05). Preoperative physiology scores were significantly lower in men with a screen-detected AAA (median 19, range 13-29 versus 21, 12-41, p<0.001). POSSUM operative scores were similar between the groups. Actual versus predicted death ratios in the sample group were more accurate using POSSUM (ratio 0.93) than P-POSSUM (2.38) analysis. CONCLUSIONS: men with a screen-detected AAA had a lower mortality rate after elective repair than in those detected incidentally; lower preoperative physiology scores suggested they were fitter (as well as younger). In this study POSSUM analysis more accurately predicted outcome than P-POSSUM. PMID- 11035971 TI - The role of type III collagen in family members of patients with abdominal aortic aneurysms. AB - OBJECTIVES: type III collagen is responsible for the tensile strength of the aorta-wall. To determine if genetic defect in the type III collagen production is associated with familial clustering of AAA. METHODS: fifty-six patients with AAA and 82 first-degree family members participated. The medical and family histories were obtained. All these relatives were screened by ultrasound for AAA. In 58 relatives of 20 families, skin biopsies were taken for protein analysis to measure type III collagen production in cultured fibroblasts. RESULTS: only one new AAA was detected in a brother of a patient. Four other relatives were already known with AAA. Three AAA patients had a type III collagen deficiency, but type III collagen was normal in all family members. CONCLUSION: type III collagen deficiency does not appear to be an aetiological factor in the development of AAA. PMID- 11035972 TI - Varicose veins on the internet. AB - OBJECTIVES: to evaluate the quality of information on the Internet concerning varicose veins. DESIGN: review of retrieved pamphlets and their scoring for educational value. MATERIALS: a sample of 41 documents were retrieved from the Internet using four "search engines". METHODS: characteristics, including country of origin, authorship, length, and presence of references were recorded. Based on factors such as disease summary, treatment options and complications a weighted score was created by two independent observers. RESULTS: eleven documents were published by an academic institution or professional organisation. Twenty-seven documents originated from private practice groups, the source of three was unidentifiable. The median weighted score was 21.5 (interquartile range: 7.5 48.5). Scores originating from non-profit making organisations were significantly higher than those from private practice groups (44.5 vs 13, p=0.04). The length of the document showed a significant positive correlation with its educational quality as measured by the information score (r=0.82, p<0.001). CONCLUSION: there is a plethora of data concerning varicose veins on the Internet. Some documents offer comprehensive information but many are confusing or misleading. Longer pamphlets and information presented by non-profit making organisations are more reliable than short documents and information offered by private medical groups. PMID- 11035973 TI - Using the Boazul cuff to reduce blood loss in varicose vein surgery. AB - OBJECTIVE: this study was designed to assess if the use of a sterile exsanguination tourniquet (Boazul cuff) reduced blood loss from the groin and avulsion wounds during varicose vein surgery, (saphenofemoral disconnection, stripping to knee and multiple avulsions). DESIGN: prospective, parallel cohort study. MATERIALS: thirty-eight patients undergoing primary varicose vein surgery. METHODS: the blood loss and number of avulsion wounds were recorded for each patient. RESULTS: the tourniquet was used on 21 legs and there were 24 legs in the non-tourniquet group. In the tourniquet group, the median blood loss from the groin was 15 ml, (range 5-70 ml), and from the leg was 5 ml, (range 0-120 ml). The corresponding figures for the group without the tourniquet were a median of 17.5 ml, (range 5-105 ml), and a median of 95 ml, (range 10-505 ml). The mean number of avulsions in the tourniquet group was 25 (range 9-38), and in the non tourniquet group was 15, (range 5-40). There was significantly less blood loss from the leg in the tourniquet group (p<0.0001) despite the fact that there were more avulsions in the tourniquet group (p<0.001). CONCLUSIONS: use of the Boazul cuff significantly decreases the blood loss from the avulsion sites during routine varicose vein surgery, and may encourage the surgeon to perform more avulsions. PMID- 11035974 TI - "Kissing balloon technique" for bilateral iliac artery obstruction in retroperitoneal fibrosis. PMID- 11035975 TI - Perivascular cystic degeneration of a saphenous vein graft. PMID- 11035976 TI - A persistent sciatic artery aneurysm with an associated internal iliac artery aneurysm. PMID- 11035977 TI - Post-traumatic clavicular pseudo-arthrosis--an unusual case of venous thoracic outlet syndrome. PMID- 11035978 TI - Coeliomesenteric trunk stenosis--a rare variation causing mesenteric ischaemia. PMID- 11035979 TI - Women's health and the environment in the 21st century. AB - For many years, the National Institute of Environmental Health Sciences has been a leader in studying the role of environmental factors in the causation of diseases that are particularly prevalent or unique to women. As we enter the next millennium, we face exciting new possibilities in broadening our understanding of how the environment impacts women's health. Sophisticated new technology and scientific information are now available to help us more precisely define environmental contributions to disease. Moreover, further development of our information base in environmental health sciences will usher in a new era of informed preventive care for women of all ages. The hallmark of this new era will be our ability to finally address the etiology and prevention of disease, rather than simply focusing on treatment and management of human illness. PMID- 11035980 TI - Advances in uterine leiomyoma research: conference overview, summary, and future research recommendations. AB - Uterine leiomyomas (fibroids, myomas) are the most common tumors occurring in the genital tract of women over 30 years of age. These benign uterine smooth-muscle tumors are estimated to be clinically significant in at least 25% of the American female population during their reproductive years. Furthermore, when thorough pathologic examination of hysterectomy specimens has been performed in patients with or without clinical history of myomatous uteri, the incidence of fibroids is 77%, suggesting that these tumors are far more prevalent than estimated by clinical cases. In spite of their high prevalence, little is known concerning the etiology or the molecular basis of their development and growth. It is well known that leiomyoma growth is regulated by ovarian steroid hormones, yet the exact molecular pathway(s) involved in tumor growth and the role of genetic susceptibility/predisposition and the environment are unclear. This article is an overview of some of the topics addressed at the conference on Women's Health and the Environment: The Next Century--Advances in Uterine Leiomyoma Research. A summary of research needs and recommendations for future research directions based on conference discussions are also presented. PMID- 11035981 TI - Genetic approaches to the study of uterine leiomyomata. AB - The use of a variety of genetic methods to dissect underlying molecular events in tumors continues to be a highly successful approach. Cytogenetic evaluations of uterine leiomyomata have revealed consistent rearrangements that have provided biological landmarks for positional cloning experiments. Genetic linkage analysis makes possible the identification of genes that might predispose women to develop myomas. New gene expression chip methods to study globally the transcripts present in uterine leiomyomata promise insight into molecular pathways in tumor biology. PMID- 11035982 TI - Pathology and pathophysiology of uterine smooth-muscle tumors. AB - Smooth-muscle tumors of uterine origin encompass a broad family of neoplasms. The leiomyoma, by far the most common of all the neoplasms, generally is hormone sensitive, with rates of growth semiquantitatively related to estrogen and progesterone receptor levels. Several forms of degenerative change can occur in the leiomyoma. The most common is hyaline degeneration, which is important in that it should not be mistaken for the coagulative tumor cell necrosis seen in leiomyosarcoma. Red degeneration (necrobiosis) is a form of degeneration that occurs characteristically but not exclusively in pregnancy, and the process is often the cause of pain and fever. Several forms of treatment have been used medically in the treatment of leiomyoma. Gonadotropin-releasing hormone analogs or agonists or selective arterial embolization with polyvinylformaldehyde particles may lead to substantial degeneration or infarction of the leiomyoma, respectively. Several variants of leiomyoma, the cellular and symplastic leiomyomas, are important to recognize, as they can be misinterpreted as sarcoma. In addition, there are two unusual growth patterns of leiomyoma that are important to recognize. Both the benign metastasizing leiomyoma and disseminated peritoneal leiomyomatosis are found outside the uterus, and neither is malignant. Recent studies offer insights into their origin and hormonal influences. From a diagnostic and therapeutic point of view, the leiomyosarcoma, while rare, is clinically of great import. Coagulative necrosis, cytologic atypia, and mitotic counts are all important in diagnosing the condition. PMID- 11035983 TI - Regulation of vascular endothelial growth factor expression by estrogens and progestins. AB - Estrogens increase the expression of vascular endothelial growth factor (VEGF) mRNA in the rodent uterus. This regulatory effect is rapid, beginning within 1 hr after hormone treatment, dose dependent, and blocked by the pure antiestrogen ICI 182,780. The induction of the transcript is blocked by inhibitors of RNA but not of protein synthesis, and we have recently identified estrogen response elements in the VEGF gene. Collectively, these findings indicate that estrogens regulate uterine VEGF expression at the transcriptional level via the classical nuclear estrogen receptor pathway. Estrogen induction of VEGF occurs in the stromal layer of the rodent uterus, and estradiol induces expression of VEGF transcript levels in cultured human uterine stromal cells. Progestins also induce VEGF expression in the rodent uterus, although the effect is less marked and slower in onset than estrogenic effects. The effect of progestins is blocked by the antiprogestin mifepristone (RU-486), suggesting that it is also mediated by a classical nuclear receptor pathway. In addition, progestins regulate expression of VEGF mRNA and protein in cultured human T47-D breast cancer cells. The development of uterine leiomyomas is associated with exposure to ovarian sex steroids, abnormal uterine bleeding is commonly seen in patients with leiomyomas, and fibroids require an increased vascular supply for their growth. These observations suggest that VEGF and other angiogenic factors may represent potential targets for the treatment and prevention of uterine fibroids. PMID- 11035984 TI - Advances in uterine leiomyoma research: the progesterone hypothesis. AB - Uterine leiomyomas are monoclonal tumors. However, the factors involved in their initiation and growth remain poorly understood. The neoplastic transformation of myometrium to leiomyoma likely involves somatic mutations of normal myometrium and the complex interactions of sex steroids and local growth factors. Traditionally, estrogen has been considered the major promoter of myoma growth. The purpose of this review is to highlight the biochemical, histologic, and clinical evidence that supports an equally important role for progesterone in the growth of uterine myomas. Biochemical studies suggest that progesterone, progestins, and the progesterone receptor modulate myoma mitotic activity. A hypothesis to explain the pathogenesis of myomas is presented. PMID- 11035985 TI - Immunohistochemical localization of growth factors and their receptors in uterine leiomyomas and matched myometrium. AB - Immunolocalization of transforming growth factor alpha (TGF-Alpha), epidermal growth factor (EGF), insulinlike growth factor (IGF)-I, vascular endothelial growth factor (VEGF(165,189,121)), basic fibroblast growth factor (FGF)-2, EGF receptor (R), IGF-IRbeta, and FGFR-1 was studied in uterine leiomyomas and matched myometrial samples taken from seven women (42-47 years of age) in the proliferative phase of the menstrual cycle. Immunolocalization of growth factor peptides was accomplished with either monoclonal or polyclonal antibodies to the amino or carboxy terminus of growth factor peptides or their respective receptors, or against full-length recombinant growth factor. All reactions were conducted using the avidin-biotin complex method. Immunolocalization of TGF alpha, EGF, EGF-R, IGF-I, IGF-IRbeta, FGF-2, FGFR-1, and VEGF was observed in the cytoplasm of smooth-muscle cells of leiomyomas and matched myometrium. The cytoplasm of vascular smooth-muscle cells expressed TGF-alpha, EGF, EGF-R, IGF-I, IGF-IRbeta, FGF-2, FGFR-1, and VEGF, whereas the vascular endothelium was positive for TGF-alpha, EGF, EGF-R, FGF-2, and FGFR-1 in both leiomyomas and matched myometria. Fibroblasts within the fibrous component of some leiomyomas were positive for IGF-I and FGF-2 and minimally positive for FGFR-1. In addition, the extracellular matrix of leiomyomas showed focal localization of FGF-2 and IGF I in some tumors. When scores of intensity and percent positive staining were compared, IGF-IRbeta was significantly increased in the leiomyomas compared to matched myometria, whereas EGF was significantly decreased in the uterine leiomyomas compared to matched myometria. In summary, these data revealed growth factors to be expressed differentially in smooth muscle, vascular and fibroblastic cell types of leiomyomas and matched myometria. Specifically, IGF IRbeta was significantly increased in leiomyomas; although a similar increase was seen with IGF-I peptide, statistical significance was not achieved. The EGF peptide was significantly decreased in the leiomyomas compared to matched myometrium. These data suggest that IGF-IRbeta and IGF-I peptide may be one of several growth factor/receptor pathways important in uterine leiomyoma growth during the proliferative phase of the menstrual cycle. In addition, decreased EGF may be secondary to the predominant estrogenic milieu present at time of sampling, as it has been proposed that progesterone, and not estrogen, may regulate EGF. PMID- 11035986 TI - Structure and function of the HMGI(Y) family of architectural transcription factors. AB - The three known members of the HMGI(Y) family of high-mobility group (HMG) mammalian nonhistone nuclear proteins (HMG-I, HMG-Y, and HMGI-C) are thought to participate in numerous biological processes (transcription, replication, retroviral integration, genetic recombination, etc.) by virtue of their ability to recognize and alter the structure of both DNA and chromatin substrates. In vitro and in vivo the HMGI(Y) proteins preferentially bind to the narrow minor groove of stretches of AT-rich DNA by means of highly conserved peptide motifs called AT hooks. In vitro the HMGI(Y) proteins also have the ability to selectively bind to distorted DNA structures and to bend, unwind, and supercoil DNA substrates. Additionally, the HMGI(Y) proteins have the ability to interact with various protein transcription factors both in vitro and in vivo. Specific protein-DNA and protein-protein interactions permit the HMGI(Y) proteins to function as architectural transcription factors that regulate gene expression in vivo by controlling the formation of stereospecific multiprotein complexes on the AT-rich regions of certain gene promoters. Transcriptional overexpression of the HMGI(Y) genes is highly correlated with both cancerous transformation and increased metastatic potential of a number of different cancers, and chromosomal rearrangements involving AT-hook motifs have been associated with various types of benign human mesenchymal tumors. The levels of HMGI(Y) proteins in human cells have been proposed to be sensitive diagnostic indicators of both neoplastic transformation and metastatic progression. Drugs based on the AT-hook motif offer the potential for development of new tumor therapeutic reagents. PMID- 11035987 TI - Comparing regulation of the connexin43 gene by estrogen in uterine leiomyoma and pregnancy myometrium. AB - Classical estrogen receptor (ER)-alpha is expressed in human myometrial and leiomyoma tissues from nonpregnant women. A comparison of these tissues shows that leiomyomas overexpress ER-alpha compared to normal myometrium. It was hypothesized that overexpression of ER-alpha in leiomyomas may account for observed overexpression of pregnancy-associated genes that are regulated by estrogen. For this reason, regulation of the labor-associated gene connexin43 cx43 was compared in primary cultures of myometrial and leiomyoma cells. It was shown that a DNA element called activating protein (AP)-1 in the cx43 promoter is necessary for induction of cx43 gene transcription in primary uterine smooth muscle cells after activation of cellular protein kinases. However, estrogen did not induce myometrial cx43 gene transcription in vitro; instead, it inhibited AP 1 induction of cx43 expression. This is likely because the myometrial and leiomyoma cells begin to express the novel ER-beta upon culturing, and agonist bound ER-beta is known to inhibit AP-1 activity. Interestingly, ER-beta is the predominant ER in myometrial tissue from pregnant women at term. Results from an examination of pregnancy myometrial tissue support the concept that AP-1 activity is involved in the induction of myometrial cx43 expression at term and that suppression of ER-beta expression is needed for this induction. As pregnancy myometrium expresses primarily ER-ss, and nonpregnancy leiomyomas express primarily ER-Alpha, AP-1 activity is predicted to be suppressed in pregnancy myometrium and elevated in leiomyomas under the influence of estrogen. This may be important in understanding tumor pathology, as AP-1 activity is associated with cell growth. PMID- 11035988 TI - Pregnancy, parturition, and prostaglandins: defining uterine leiomyomas. AB - Leiomyomas, benign smooth muscle tumors of the uterus, are the most common gynecologic neoplasm in women. Studies with surgically resected human tissues and primary cultures have revealed that several genes are differentially expressed in leiomyomas compared to matched normal myometrium. An estrogen-driven pattern of gene expression in leiomyomas, similar to that seen in normal myometrium during pregnancy and parturition, is associated with a persistent inappropriate response of neoplastic myometrial smooth muscle cells to ovarian hormones. This is possibly due to aberrant expression levels or signaling via estrogen and progesterone receptors. We propose the hypothesis that uterine leiomyomas mimic a differentiated myometrial cell at pregnancy and exhibit a hypersensitivity to sex steroid hormones that prevents the cells from responding to normal apoptotic or dedifferentiation signals and from returning to a nongravid phenotype. Support of this hypothesis is derived from experimental studies in female Eker rats that develop uterine leiomyomas with many similarities to the human disease. Our hypothesis accounts for the benign nature of these tumors and their high incidence in women during the reproductive years. By identifying the factors that participate in parturition and involution of the pregnant myometrium, we may better define uterine leiomyomas and thus identify novel targets for therapeutic strategies to treat these tumors. PMID- 11035989 TI - Epidemiologic contributions to understanding the etiology of uterine leiomyomata. AB - Uterine leiomyomata are hormonally dependent tumors that are a major source of gynecologic morbidity among women of reproductive age. Relatively few studies have attempted to identify specific risk factors for these neoplasms. In this review of epidemiologic contributions to the etiology of uterine leiomyomata, we begin by outlining methodologic issues in epidemiologic studies that arise from the fact that a large proportion of uterine leiomyomata does not come to medical attention. We then review the major findings from published epidemiologic studies, which to date have focused primarily on menstrual and childbearing history, exogenous hormone use, obesity, cigarette smoking, and other lifestyle and behavioral characteristics that may in part reflect aspects of a woman's hormonal milieu. None of the potential risk factors studied have demonstrated sufficiently consistent associations to guide decisions on the primary prevention of uterine leiomyomata. Clarifying the etiology and natural history of uterine leiomyomata will require studies designed to address methodologic issues and test hypotheses involving environmental and lifestyle influences on endocrine function, as well as on other possible etiologic mechanisms. Recent advances in molecular genetics present opportunities for epidemiologic studies of uterine leiomyomata to incorporate biomarkers of somatic changes found in these tumors and to examine inherited genetic factors related to possible causal physiologic mechanisms. PMID- 11035990 TI - Influence of exogenous estrogen receptor ligands on uterine leiomyoma: evidence from an in vitro/in vivo animal model for uterine fibroids. AB - The remarkable frequency of uterine leiomyoma in the human population calls into question the potential for the participation of environmental factors in tumor etiology. Having been implicated in the dramatic rise in hormone-related cancers in recent years, endocrine disruptors are salient suspects in this pathogenesis, although the mechanism by which they might participate is unclear. Investigations using the Eker rat model show that uterine leiomyoma may have an enhanced sensitivity to modulation via the estrogen receptor. This sensitivity could make these tumors a target for disruption by exogenous estrogen receptor ligands. Direct evidence for a pathogenic role of exogenous compounds in leiomyomas is lacking; however, it can be demonstrated that such diverse agents as organochlorine pesticides, dietary flavonoids, botanical extracts, and therapeutic antiestrogens have either estrogen agonist or antagonist function in myometrial tissues. The use of this model will help define the impact of exogenous estrogen receptor modulators on uterine leiomyoma and will permit the evaluation of strategies for therapeutic intervention. PMID- 11035991 TI - Clinical decision making regarding leiomyomata: what we need in the next millenium. AB - Leiomyomata represent the most common gynecologic tumors and are responsible for over 200,000 hysterectomies per year. They are almost invariably benign and represent clonal expansion of individual myometrial cells. They can cause a variety of symptoms including menometrorrhagia, dysmenorrhea, pelvic pain, reproductive failure, and compression of adjacent pelvic viscera, or be totally asymptomatic. Leiomyomata are more common in African-American women and have a non-Mendelian inheritance pattern with up to a 50% recurrence rate after surgical removal. The therapeutic choices depend on the goals of therapy, with hysterectomy most often used for definitive treatment, and myomectomy when preservation of childbearing is desired. Intracavitary and submucous leiomyomata can be removed by hysteroscopic resection. Laparoscopic myomectomy is now technically possible but apparently with an increased risk of uterine rupture during pregnancy. Although gonadotropin-releasing hormone-agonist-induced hypogonadism can reduce the volume of leiomyomata, the severe side effects and prompt recurrences make them useful only for short-term goals such as reversing anemia or shrinking an intracavitary tumor prior to hysteroscopic resection. Nonextirpative approaches such as myolysis and uterine artery embolization are being evaluated, and may provide more options if they prove to be safe and efficacious in long-term follow-up. Ultimately, if the genetic basis for fibroid development and/or the molecular mechanism(s) of myometrial proliferation are understood, additional nonsurgical therapeutic interventions may be forthcoming. Current clinical needs are to a) determine an effective prevention strategy in genetically predisposed individuals; b) slow the growth of leiomyomata; c) identify the mechanisms of infertility; d) improve early detection; e) develop better surgical techniques; f) reduce recurrences after myomectomy; g) develop nonextirpative options; and h) evaluate their long-term results. PMID- 11035992 TI - Review of the evidence for treatment of leiomyomata. AB - Leiomyomata of the uterus are common tumors of women, and a number of treatments are now available for symptoms of this disorder. Surgical treatments include hysterectomy, abdominal myomectomy, laparoscopic myomectomy, and myolysis. Nonsurgical treatments include uterine artery embolization and medical therapy. All have a small amount of accumulated data, but existing studies are generally small and of poor quality. There is a strong need for appropriately designed and analyzed randomized clinical trials, and surgical trials should preferably be multicenter/multisurgeon. In summary, although a large array of options exist, there are still little data upon which to base decisions regarding optimal treatment approaches. PMID- 11035993 TI - Impact of uterine fibroids on ART outcome. AB - Our objective was to investigate the effect of subserosal (SS), intramural (IM), and submucosal (SM) fibroids on the outcome of assisted reproductive technology (ART) treatment. A retrospective comparative study at a tertiary referral center for infertility was designed. The treatment outcome of 106 ART cycles in 88 patients with uterine fibroids (33 SS, 46 IM without cavity distortion, 9 SM) was compared with that of 318 ART cycles in age-matched patients without fibroids. The main outcome measure(s) were the findings on transvaginal uterine ultrasonography performed before the initiation of treatment and pregnancy and implantation rates. The pregnancy rates per transfer were 34.1, 16.4, 10, and 30.1% in the patients with SS fibroids, IM fibroids, SM fibroids, and no fibroids, respectively. The implantation rates were 15.1, 6.4, 4.3, and 15.7%, respectively. Both rates were significantly lower in patients with IM fibroids than in those with SS fibroids or no fibroids. We conclude that pregnancy and implantation rates were significantly lower in the groups of patients with IM and SM fibroids, even when there was no deformation of the uterine cavity. Pregnancy and implantation rates were not influenced by the presence of SS fibroids. Surgical or medical treatment should be considered in infertile patients who have IM and/or SM fibroids before resorting to ART treatment. PMID- 11035994 TI - Novel therapeutic strategies for leiomyomas: targeting growth factors and their receptors. AB - Leiomyomas (fibroids) are benign smooth-muscle cell (SMC) tumors of the uterus and are the most common pelvic tumors in women. These tumors occur primarily during the reproductive years and are the most common indication for hysterectomy in women. Unfortunately the only effective treatments for leiomyomas and the associated abnormal uterine bleeding are surgical, involving either hysterectomy, myomectomy, or hysteroscopic removal of the tumors. The goal of this paper is to discuss recent research findings that support the idea of using therapeutic compounds that block the actions of specific growth factors as therapeutic agents for treatment of leiomyomas and abnormal uterine bleeding. Most of the studies were carried out using cell cultures of leiomyoma or myometrial SMCs. Primary cultures of SMCs provide a system for investigation of the roles of growth factors and their receptors in proliferation of normal myometrial and leiomyoma SMCs. Several growth factors have been shown to be present and to have regulatory roles in the proliferation of uterine SMCs. Bioassay and Western blotting of fast protein liquid chromatography fractions of tissue extracts identified platelet derived growth factor, heparin-binding epidermal growth factor, hepatoma-derived growth factor, and basic fibroblast growth factor in normal myometrium and fibroid tumors. The presence of heparin-binding growth factors suggests a possible focus for therapeutic agents. RG13577 (a heparinlike compound) and halofuginone (an alkyloid) reversibly inhibited DNA synthesis of normal myometrial and leiomyoma cells without toxic effects. Pirfenidone, a known antifibrotic drug, inhibited DNA synthesis and synthesis of collagen type I mRNA in normal and fibroid cells, and decreased collagen type III mRNA only in normal myometrial cells. Another hopeful therapeutic candidate, interferon-Alpha, significantly inhibited growth factor-stimulated proliferation in both normal and leiomyoma cells. These results suggest that future nonsurgical treatments for leiomyomas may include compounds that block the actions of specific growth factors that regulate proliferation and collagen production by uterine SMCs. PMID- 11035995 TI - Current directions in physiological modeling for environmental health sciences: an overview. PMID- 11035996 TI - The multiple indicator dilution method and its utility in risk assessment. AB - The multiple-indicator dilution (MID) technique entails the injection of a mixture of labeled indicators into the blood vessel immediately at the entrance of an organ, e.g., the liver, kidney, heart, or lung, and characterization of outflow dilution profiles from timed venous samples. The mathematical basis of the method encompasses linear systems of partial differential equations that are formulated for flow- or barrier-limited transport combined with intracellular metabolism/excretion. The concept can be generalized to include metabolites. MID experiments are useful for determining tissue partition coefficients as well as kinetic parameters such as membrane permeabilities or metabolic/excretory intrinsic clearances, factors that affect the mean residence times or exposure of solutes to the organ. The main utility of the MID method lies in its role in identifying the basic mechanisms of the interaction of organs with vascular components. The concentration dependence in transport and removal is revealed by the rate coefficients upon varying the input concentrations of unlabeled substances into the organ at steady state. The data obtained with MID experiments can be incorporated into physiologically based pharmacokinetic (PBPK) models such as those used for biological risk assessment. This is especially pertinent in the case where diffusional barriers appear within organs. The insight gained from the MID organ approach may be useful for PBPK models with more realistic representation of organ kinetics. PMID- 11035997 TI - A model of gonadotropin regulation during the menstrual cycle in women: qualitative features. AB - Increasing concerns that environmental contaminants may disrupt the endocrine system require development of mathematical tools to predict the potential for such compounds to significantly alter human endocrine function. The endocrine system is largely self-regulating, compensating for moderate changes in dietary phytoestrogens (e.g., in soy products) and normal variations in physiology. However, severe changes in dietary or oral exposures or in health status (e.g., anorexia), can completely disrupt the menstrual cycle in women. Thus, risk assessment tools should account for normal regulation and its limits. We present a mathematical model for the synthesis and release of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) in women as a function of estrogen, progesterone, and inhibin blood levels. The model reproduces the time courses of LH and FSH during the menstrual cycle and correctly predicts observed effects of administered estrogen and progesterone on LH and FSH during clinical studies. The model should be useful for predicting effects of hormonally active substances, both in the pharmaceutical sciences and in toxicology and risk assessment. PMID- 11035998 TI - Statistical issues in toxicokinetic modeling: a bayesian perspective. AB - Determining the relationship between an exposure and the resulting target tissue dose is a critical issue encountered in quantitative risk assessment (QRA). Classical or physiologically based toxicokinetic (PBTK) models can be useful in performing that task. Interest in using these models to improve extrapolations between species, routes, and exposure levels in QRA has therefore grown considerably in recent years. In parallel, PBTK models have become increasingly sophisticated. However, development of a strong statistical foundation to support PBTK model calibration and use has received little attention. There is a critical need for methods that address the uncertainties inherent in toxicokinetic data and the variability in the human populations for which risk predictions are made and to take advantage of a priori information on parameters during the calibration process. Natural solutions to these problems can be found in a Bayesian statistical framework with the help of computational techniques such as Markov chain Monte Carlo methods. Within such a framework, we have developed an approach to toxicokinetic modeling that can be applied to heterogeneous human or animal populations. This approach also expands the possibilities for uncertainty analysis. We present a review of these efforts and other developments in these areas. Appropriate statistical treatment of uncertainty and variability within the modeling process will increase confidence in model results and ultimately contribute to an improved scientific basis for the estimation of occupational and environmental health risks. PMID- 11035999 TI - Canonical modeling: review of concepts with emphasis on environmental health. AB - The article reviews concepts of canonical modeling in the context of environmental health. Based on biochemical systems theory, the canonical approach was developed over the past thirty years and applied to complex systems primarily in biochemistry and the regulation of gene expression. Canonical modeling is based on nonlinear ordinary differential equations whose right-hand sides consist of products of power-law functions. This structure results from the linearization of complex processes in logarithmic space. The canonical structure has many intriguing features. First, almost any system of smooth functions or ordinary differential equations can be recast equivalently in a canonical model, which demonstrates that the model structure is rich enough to deal with all relevant nonlinearities. Second, a large body of successful applications suggests that canonical models are often valid and accurate representations of quite complex, real-world systems. Third, a set of guidelines supports the modeler in all phases of analysis. These guidelines address model design, algebraic and numerical analysis, and the interpretation of results. Fourth, the structure of canonical models, especially those in S-system form, facilitates algebraic and numerical analyses. Of particular importance is the derivation of steady-state solutions in an explicit symbolic or numerical form, which allows further assessments of stability and robustness. The homogeneous structure of canonical models has also led to the development of very efficient, customized computer algorithms for all steps of a typical analysis. Fifth, a surprising number of models currently used in environmental health research are special cases of canonical models. The traditional models are thus subsumed in one modeling framework, which offers new avenues of analysis and interpretation. PMID- 11036000 TI - The privileged access model of 1,3-butadiene disposition. AB - In previous attempts to model disposition of 1,3-butadiene in mice and rats, parameter values for 1,2-epoxybut-3-ene metabolism were optimized to reproduce elimination of this gas from closed chambers. However, each of these models predicted much higher concentrations of circulating epoxybutene than were subsequently measured in animals exposed to butadiene. To account for this discrepancy, a previous physiologically based pharmacokinetic model of butadiene disposition was modified to describe a transient complex between cytochrome P450 and epoxide hydrolase on the endoplasmic reticulum membrane. In this model the epoxide products are directly transferred from the P450 to the epoxide hydrolase in competition with release of products into the cytosol. The model includes flow restricted delivery of butadiene and epoxides to gastrointestinal tract, liver, lung, kidney, fat, other rapidly perfused tissues, and other slowly perfused tissues. Blood was distributed among compartments for arterial, venous, and capillary spaces. Oxidation of butadiene and epoxybutene and hydrolysis and glutathione conjugation of epoxides were included in liver, lung, and kidney. The model reproduces observed uptake of butadiene and epoxybutene from closed chambers by mice and rats and steady-state concentrations of butadiene, epoxybutene, and 1,2;3,4-diepoxybutane concentrations in blood of mice and rats exposed by nose only. Successful replication of these observations indicates that the proposed privileged access of epoxides formed in situ to epoxide hydrolase is a plausible mechanistic representation for the metabolic clearance of epoxide forming chemicals. PMID- 11036001 TI - Simulation modeling of the tissue disposition of formaldehyde to predict nasal DNA-protein cross-links in Fischer 344 rats, rhesus monkeys, and humans. AB - Formaldehyde inhalation causes formation of DNA-protein cross-links (DPX) in the nasal mucosa of Fischer 344 (F344) rats and rhesus monkeys. DPX are considered to be part of the mechanism by which cytotoxic and carcinogenic effects of formaldehyde in laboratory animals are exerted, and DPX data have been used as a measure of tissue dose in cancer risk assessments for formaldehyde. Accurate prediction of DPX concentrations in humans is therefore desirable. The goal of this work was to increase confidence in the prediction of human DPX by refining earlier models of formaldehyde disposition and DPX kinetics in the nasal mucosa. Anatomically accurate, computational fluid dynamics models of the nasal airways of F344 rats, rhesus monkeys, and humans were used to predict the regional flux of formaldehyde to the respiratory and olfactory mucosa. A previously developed model of the tissue disposition of formaldehyde and of DPX kinetics was implemented in the graphical simulation tool SIMULINK and linked to the regional flux predictions. Statistical optimization was used to identify parameter values, and good simulations of the data were obtained. The parameter estimates for rats and monkeys were used to guide allometric scale-up to the human case. The relative levels of nasal mucosal DPX in rats, rhesus monkeys, and humans for a given inhaled concentration of formaldehyde were predicted by the model to vary with concentration. This modeling approach reduces uncertainty in the prediction of human nasal mucosal DPX resulting from formaldehyde inhalation. PMID- 11036003 TI - Fluconazole plus cyclosporine: a fungicidal combination effective against experimental endocarditis due to Candida albicans. AB - Recent observations demonstrated that fluconazole plus cyclosporine (Cy) synergistically killed Candida albicans in vitro. This combination was tested in rats with C. albicans experimental endocarditis. The MICs of fluconazole and Cy for the test organism were 0.25 and >10 mg/liter, respectively. Rats were treated for 5 days with either Cy, amphotericin B, fluconazole, or fluconazole-Cy. Although used at high doses, the peak concentrations of fluconazole in the serum of rats (up to 4.5 mg/liter) were compatible with high-dose fluconazole therapy in humans. On the other hand, Cy concentrations in serum (up to 4.5 mg/liter) were greater than recommended therapeutic levels. Untreated rats demonstrated massive pseudohyphal growth in both the vegetations and the kidneys. However, only the kidneys displayed concomitant polymorphonuclear infiltration. The therapeutic results reflected this dissociation. In the vegetations, only the fungicidal fluconazole-Cy combination significantly decreased fungal densities compared to all groups, including amphotericin B (P < 0.0001). In the kidneys, all regimens except the Cy regimen were effective, but fluconazole-Cy remained superior to amphotericin B and fluconazole alone in sterilizing the organs (P < 0.0001). While the mechanism responsible for the fluconazole-Cy interaction is hypothetical, this observation opens new perspectives for fungicidal combinations between azoles and other drugs. PMID- 11036002 TI - A neutrophil-derived anti-infective molecule: bactericidal/permeability increasing protein. PMID- 11036004 TI - Anti-human immunodeficiency virus activities of nucleosides and nucleotides: correlation with molecular electrostatic potential data. AB - Examination of the anti-human immunodeficiency virus (HIV) data of some normal and isomeric dideoxynucleosides (ddNs and isoddNs), their three-dimensional (3-D) electron density patterns, their electrostatic potential surfaces (EPS), and their conformational maps reveals some interesting correlations. For example, the EPS of (S,S)-isoddA shows regions of high and low electrostatic potential remarkably similar to those of beta-D-3'-azido-3'-deoxythymidine (beta-D-AZT), ( )-oxetanocin A, and (-)-carbovir. Such correlations involving EPS data and anti HIV activity were also found with many other active nucleosides. Conversely, inactive compounds had EPS different from those of compounds in the same series that were active. For example, apio-ddNs, which are inactive against HIV, exhibit clear differences in electrostatic potential and 3-D electron density shape from isoddNs that are active against HIV. Additionally, the inactivity of (S,S)-isoddC and (S,S)-isoddT can be correlated convincingly with a combination of their EPS data and their conformational energy maps. The electrostatic potential distributions of active nucleoside triphosphates show remarkable correlations. For example, (S,S)-isoddATP, AZT triphosphate (AZTTP), and oxetanocin A TP have similar 3-D electron density surface patterns and similar high and low regions of electrostatic potential, which may suggest that these compounds proceed through related mechanisms in their interactions with, and inhibition of, HIV reverse transcriptase (RT). Docking of AZTTP, (S,S)-isoddATP, and other active triphosphates into the active site of HIV RT and calculation of the EPS of both the nucleotide and the active site show that there is excellent matching between inhibitor and enzyme binding site EPS data. The structure-activity profile discovered has contributed to the development of a first predictive quantitative structure-activity relationship analysis in the area. PMID- 11036005 TI - Once-daily dosing in dogs optimizes daptomycin safety. AB - Daptomycin is a novel lipopeptide antibiotic with potent bactericidal activity against most clinically important gram-positive bacteria, including resistant strains. Daptomycin has been shown to have an effect on skeletal muscle. To guide the clinical dosing regimen with the potential for the least effect on skeletal muscle, two studies were conducted with dogs to compare the effects of repeated intravenous administration every 24 h versus every 8 h for 20 days. The data suggest that skeletal-muscle effects were more closely related to the dosing interval than to either the maximum concentration of the drug in plasma or the area under the concentration-time curve. Both increases in serum creatine phosphokinase activity and the incidence of myopathy observed at 25 mg/kg of body weight every 8 h were greater than those observed at 75 mg/kg every 24 h despite the lower maximum concentration of drug in plasma. Similarly, the effects observed at 25 mg/kg every 8 h were greater than those observed at 75 mg/kg every 24 h at approximately the same area under the concentration-time curve from 0 to 24 h. Once-daily administration appeared to minimize the potential for daptomycin related skeletal-muscle effects, possibly by allowing for more time between doses for repair of subclinical effects. Thus, these studies with dogs suggest that once-daily dosing of daptomycin in humans should have the potential to minimize skeletal-muscle effects. In fact, interim results of ongoing clinical trials, which have focused on once-daily dosing, appear to be consistent with this conclusion. PMID- 11036006 TI - Contribution of dithiol ligands to in vitro and in vivo trypanocidal activities of dithiaarsanes and investigation of ligand exchange in an aqueous solution. AB - Twelve new dithiaarsanes were evaluated for their in vitro and in vivo trypanocidal properties in regard to their three parent molecules, 4-amino phenylarsenoxide, melarsenoxide, and 4-dansylamino-phenylarsenoxide. The most potent dithiaarsane, compound 2b, had a minimum effective concentration of 1.5 nM after 48 h of incubation and at a dose of 0.39 micromol/kg of body weight (0.2 mg/kg) administered subcutaneously cured 100% of mice acutely infected with Trypanosoma brucei brucei CMP. With this model, the chemotherapeutic index of compound 2b was 512, compared to 256 for melarsamine dihydrochloride (Cymelarsan) under the same conditions. With a chronic infection produced by T. brucei brucei GVR, compound 2b cured 100% of mice after treatment at a dose of 25 micromol/kg (12.5 mg/kg) for 4 consecutive days, whereas melarsamine dihydrochloride and potassium melarsonyl (Trimelarsan) cured less than 50% mice at this dose. For both acute and late-stage infections, dithiaarsanes having a melaminophenyl ring exhibited the most-potent trypanocidal activity. Compound 2b is thus one of the most active organoarsenicals described in a mouse trypanosomiasis model. Considering that the main intracellular targets of organoarsenicals are thiol groups, we studied the possibility of ligand exchange between Cymelarsan and several dithiols. In aqueous solution, we observed a rapid exchange of cysteamine from melarsamine with free cysteamine and also with various dithiols always in favor of more stable cyclic derivatives. These ligand exchanges suggest the ability of trivalent organoarsenicals to react with targets such as trypanothione and dihydrolipoic acid. Among several ligands, a 1,3-dimercaptopropane moiety appeared the most suitable for trypanocidal activity. PMID- 11036007 TI - Activities of clinafloxacin, gatifloxacin, gemifloxacin, and trovafloxacin against recent clinical isolates of levofloxacin-resistant Streptococcus pneumoniae. AB - The activities of two investigational fluoroquinolones and three fluoroquinolones that are currently marketed were determined for 182 clinical isolates of Streptococcus pneumoniae. The collection included 57 pneumococcal isolates resistant to levofloxacin (MIC >/= 8 microg/ml) recovered from patients in North America and Europe. All isolates were tested with clinafloxacin, gatifloxacin, gemifloxacin, levofloxacin, and trovafloxacin by the National Committee for Clinical Laboratory Standards broth microdilution and disk diffusion susceptibility test methods. Gemifloxacin demonstrated the greatest activity on a per gram basis, followed by clinafloxacin, trovafloxacin, gatifloxacin, and levofloxacin. Scatterplots of the MICs and disk diffusion zone sizes revealed a well-defined separation of levofloxacin-resistant and -susceptible strains when the isolates were tested against clinafloxacin and gatifloxacin. DNA sequence analyses of the quinolone resistance-determining regions of gyrA, gyrB, parC, and parE from 21 of the levofloxacin-resistant strains identified eight different patterns of amino acid changes. Mutations among the four loci had the least effect on the MICs of gemifloxacin and clinafloxacin, while the MICs of gatifloxacin and trovafloxacin increased by up to six doubling dilutions. These data indicate that the newer fluoroquinolones have greater activities than levofloxacin against pneumococci with mutations in the DNA gyrase or topoisomerase IV genes. Depending upon pharmacokinetics and safety, the greater potency of these agents could provide improved clinical efficacy against levofloxacin-resistant pneumococcal strains. PMID- 11036008 TI - Stages of polymyxin B interaction with the Escherichia coli cell envelope. AB - The effects of polymyxin B (PMB) on the Escherichia coli outer (OM) and cytoplasmic membrane (CM) permeabilities were studied by monitoring the fluxes of tetraphenylphosphonium, phenyldicarbaundecaborane, and K(+) and H(+) ions. At concentrations between 2 and 20 microgram/ml, PMB increased the OM permeability to lipophilic compounds and induced a leakage of K(+) from the cytosol and an accumulation of lipophilic anions in the cellular membranes but did not cause the depolarization of the CM. At higher concentrations, PMB depolarized the CM, forming ion-permeable pores in the cell envelope. The permeability characteristics of PMB-induced pores mimic those of bacteriophage- and/or bacteriocin-induced channels. However, the bactericidal effect of PMB took place at concentrations below 20 microgram/ml, indicating that this effect is not caused by pore formation. Under conditions of increased ionic strength, PMB made the OM permeable to lipophilic compounds and decreased the K(+) gradient but was not able to depolarize the cells. The OM-permeabilizing effect of PMB can be diminished by increasing the concentration of Mg(2+). The major new findings of this work are as follows: (i) the OM-permeabilizing action of PMB was dissected from its depolarizing effect on the CM, (ii) the PMB-induced ion-permeable pores in bacterial envelope were registered, and (iii) the pore formation and depolarization of the CM are not obligatory for the bactericidal action of PMB and dissipation of the K(+) gradient on the CM. PMID- 11036009 TI - Genetic diversity of the tet(M) gene in tetracycline-resistant clonal lineages of Streptococcus pneumoniae. AB - The aim of the present study was to examine the stability and evolution of tet(M) mediated resistance to tetracyclines among members of different clonal lineages of Streptococcus pneumoniae. Thirty-two tetracycline-resistant isolates representing three national (Spanish serotype 14, Spanish serotype 15, and Polish serotype 23F) and one international (Spanish serotype 23F) multidrug-resistant epidemic clones were all found to be tet(M) positive and tet(O), tet(K), and tet(L) negative. These isolates all carried the integrase gene, int, which is associated with the Tn1545-Tn916 family of conjugative transposons. High resolution restriction analysis of tet(M) products identified six alleles, tet(M)1 to tet(M)6: tet(M)1 to tet(M)3 and tet(M)5 in isolates of the Spanish serotype 14 clone, tet(M)4 in both the Spanish serotype 15 and 23F clones, and tet(M)6, the most divergent allele, in the Polish 23F clone. This indicates that tet(M) variation can occur at the inter- and intraclone levels in pneumococci. Two alleles of int were identified, with int1 being found in all isolates apart from members of the international Spanish 23F clone, which carried int2. Susceptibility to tetracycline, doxycycline, and minocycline was evaluated for all isolates with or without preincubation in the presence of subinhibitory concentrations of tetracyclines. Resistance to tetracyclines was found to be inducible in isolates of all clones; however, the strongest induction was observed in the Spanish serotype 15 and 23F clones carrying tet(M)4. Tetracycline was found to be the strongest inducer of resistance, and minocycline was found to be the weakest inducer of resistance. PMID- 11036010 TI - Genetic analysis of azole resistance in the Darlington strain of Candida albicans. AB - High-level azole resistance in the Darlington strain of Candida albicans was investigated by gene replacement in C. albicans and expression in Saccharomyces cerevisiae. We sequenced the ERG11 gene, which encodes the sterol C(14)alpha demethylase, from our copy of the Darlington strain. Both alleles contained the histidine for tyrosine substitution at position 132 (Y132H) reported in Darlington by others, but we also found a threonine-for-isoleucine substitution (I471T) not previously reported in the C. albicans ERG11. The encoded I471T change in amino acids conferred azole resistance when overexpressed alone and increased azole resistance when added to the Y132H amino acid sequence in an S. cerevisiae expression system. Replacement of one copy of ERG11 in an azole susceptible strain of C. albicans with a single copy of the Darlington ERG11 resulted in expression of the integrated copy and a modest increase in azole resistance. The profound azole resistance of the Darlington strain is the result of multiple mutations. PMID- 11036011 TI - Activities of LY333328 and vancomycin administered alone or in combination with gentamicin against three strains of vancomycin-intermediate Staphylococcus aureus in an in vitro pharmacodynamic infection model. AB - Staphylococcus aureus with intermediate glycopeptide susceptibility (glycopeptide intermediate S. aureus [GISA]) has been isolated from patients with apparent therapy failures. We studied the killing activity of vancomycin over a range of simulated conventional doses (1 to 1.5 g every 12 h) against three of these GISA strains in an in vitro pharmacodynamic infection model. We also studied the activity of a new glycopeptide (LY333328) at a simulated dose of 3 mg/kg of body weight every 24 h or 5 mg/kg every 24 h, as well as the potential for vancomycin and gentamicin synergy against these GISA strains. Four doses of vancomycin with or without gentamicin or two doses of LY333328 were administered over the 48-h study period. The vancomycin and LY333328 MICs and minimal bactericidal concentrations (MBCs) for the three GISA strains (strains 14379, 992, and Mu50) were 8 and 8 microgram/ml and 1 and 2 microgram/ml, respectively, for GISA 14379, 6 and 6 microgram/ml and 1 and 1 microgram/ml, respectively, for GISA 992, and 8 and 12 microgram/ml and 2 and 8 microgram/ml, respectively, for GISA Mu50. Vancomycin and LY333328 MICs and MBCs were 0.75 and 1.0 microgram/ml and 1 and 1 microgram/ml, respectively for a vancomycin-susceptible comparator strain (methicillin-resistant S. aureus [MRSA] 494). The addition of albumin to the growth medium increased the LY333328 MICs and MBCs approximately 8- to 16-fold. Vancomycin was bacteriostatic against the three GISA strains at doses of 1, 1.125, and 1.25 g every 12 h. Vancomycin was bactericidal at the dose of 1.5 g every 12 h against all strains; bactericidal activity occurred against the GISA strains at 8- to 10-fold lower ratios of the peak concentration to the MIC and the area under the concentration-time curve from time zero to 24 h (AUC(0-24)) to the MIC compared to those for the vancomycin-sensitive control strain. Overall, vancomycin activity was significantly correlated with the AUC(0-24) (R(2) = 0.79; P < 0.001) by multiple stepwise regression analyses. The addition of gentamicin did not significantly affect killing activity against any strain. LY333328 was bactericidal against GISA strains 14379 and 992 and against MRSA 494 only with the 5-mg/kg/day dose simulations. The higher dose of LY333328 also prevented regrowth over the 48-h experiments for all strains tested. Higher doses of vancomycin (1.5 g every 12 h) and LY333328 (5 mg/kg every 24 h) may represent potential treatment options for infections caused by GISA strains. PMID- 11036012 TI - In vitro activity of the new ketolide telithromycin compared with those of macrolides against Streptococcus pyogenes: influences of resistance mechanisms and methodological factors. AB - One hundred and seven clinical isolates of Streptococcus pyogenes, 80 susceptible to macrolides and 27 resistant to erythromycin A (MIC >0.5 microgram/ml), were examined. The erythromycin A-lincomycin double-disk test assigned 7 resistant strains to the M-phenotype, 8 to the inducible macrolide, lincosamide, and streptogramin B resistance (iMLS(B)) phenotype, and 12 to the constitutive MLS(B) resistance (cMLS(B)) phenotype. MICs of erythromycin A, clarithromycin, azithromycin, roxithromycin, and clindamycin were determined by a broth microdilution method. MICs of telithromycin were determined by three different methods (broth microdilution, agar dilution, and E-test methods) in an ambient air atmosphere and in a 5 to 6% CO(2) atmosphere. Erythromycin A resistance genes were investigated by PCR in the 27 erythromycin A-resistant isolates. MICs of erythromycin A and clindamycin showed six groups of resistant strains, groups A to F. iMLS(B) strains (A, B, and D groups) are characterized by two distinct patterns of resistance correlated with genotypic results. A- and B-group strains were moderately resistant to 14- and 15-membered ring macrolides and highly susceptible to telithromycin. All A- and B-group isolates harbored erm TR gene, D group strains, highly resistant to macrolides and intermediately resistant to telithromycin (MICs, 1 to 16 microgram/ml), were all characterized by having the ermB gene. All M-phenotype isolates (C group), resistant to 14- and 15-membered ring macrolides and susceptible to clindamycin and telithromycin, harbored the mefA gene. All cMLS(B) strains (E and F groups) with high level of resistance to macrolides, lincosamide, and telithromycin had the ermB gene. The effect of 5 to 6% CO(2) was remarkable on resistant strains, by increasing MICs of telithromycin from 1 to 6 twofold dilutions against D-E- and F-group isolates. PMID- 11036013 TI - Purification and biochemical characterization of the VIM-1 metallo-beta lactamase. AB - VIM-1 is a new group 3 metallo-beta-lactamase recently detected in carbapenem resistant nosocomial isolates of Pseudomonas aeruginosa from the Mediterranean area. In this work, VIM-1 was purified from an Escherichia coli strain carrying the cloned bla(VIM-1) gene by means of an anion-exchange chromatography step followed by a gel permeation chromatography step. The purified enzyme exhibited a molecular mass of 26 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and an acidic pI of 5.1 in analytical isoelectric focusing. Amino-terminal sequencing showed that mature VIM-1 results from the removal of a 26-amino-acid signal peptide from the precursor. VIM-1 hydrolyzes a broad array of beta-lactam compounds, including penicillins, narrow- to expanded-spectrum cephalosporins, carbapenems, and mechanism-based serine-beta-lactamase inactivators. Only monobactams escape hydrolysis. The highest catalytic constant/K(m) ratios (>10(6) M(-1). s(-1)) were observed with carbenicillin, azlocillin, some cephalosporins (cephaloridine, cephalothin, cefuroxime, cefepime, and cefpirome), imipenem, and biapenem. Kinetic parameters showed remarkable variability with different beta-lactams and also within the various penam, cephem, and carbapenem compounds, resulting in no clear preference of the enzyme for any of these beta-lactam subfamilies. Significant differences were observed with some substrates between the kinetic parameters of VIM-1 and those of other metallo-beta-lactamases. Inactivation assays carried out with various chelating agents (EDTA, 1,10-o-phenanthroline, and pyridine-2,6-dicarboxylic acid) indicated that formation of a ternary enzyme-metal-chelator complex precedes metal removal from the zinc center of the protein and revealed notable differences in the inactivation parameters of VIM-1 with different agents. PMID- 11036015 TI - In vitro efficacy of nikkomycin Z against the human isolate of the microsporidian species Encephalitozoon hellem. AB - Since 1985 microsporidia have been recognized as a cause of emerging infections in humans, mainly in immunocompromised human immunodeficiency virus-positive subjects. As chitin is a basic component of the microsporidian infective stage, the spore, we evaluated in vitro the susceptibility of a human-derived strain of Encephalitozoon hellem to nikkomycin Z, a peptide-nucleoside antibiotic known as a competitive inhibitor of chitin synthase enzymes. Transmission electron microscopy showed that this drug, at 25 microgram/ml, reduced the number of parasitic foci by about 35% +/- standard deviation after 7 days of culture (P < 0.0001) and induced cell damage of both mature and immature spores and also other sporogonic and merogonic stages. In particular, an irregular outline of the cell shape and an abnormally condensed cytoplasm in meronts and sporonts were documented. Also, the polar tubule and the polaroplast membranes appeared disarrayed in the sporoblast stage. The spore wall showed an enlarged endospore and delaminated exospore. Mature spores had a complete cytoplasmic disorganization and a swollen and delaminated cell wall. No ultrastructural cell damage was observed in uninfected control cultures treated with the drug. PMID- 11036014 TI - A comparative In vitro surveillance study of gemifloxacin activities against 2,632 recent Streptococcus pneumoniae isolates from across Europe, North America, and South America. The Gemifloxacin Surveillance Study Research Group. AB - From 1997 to 1999, 94 study centers in 15 European, 3 North American, and 2 South American countries contributed 2,632 isolates of Streptococcus pneumoniae to an international antimicrobial susceptibility testing study. Only 62.0% of isolates were susceptible to penicillin, while 22.3% were penicillin intermediate and 15.6% were penicillin resistant. Resistance to trimethoprim-sulfamethoxazole (24.4%), azithromycin (26.0%), and clarithromycin (27.1%) was also highly prevalent. For the penicillin-resistant isolates (n = 411), the MICs at which 90% of isolates are inhibited (MIC(90)s) for gemifloxacin, levofloxacin, ofloxacin, clarithromycin, and azithromycin were 0.03, 1, 2, >16, and >64 microgram/ml, respectively. Similarly, for isolates resistant to both azithromycin and clarithromycin (n = 649), gemifloxacin, levofloxacin, ofloxacin, and penicillin MIC(90)s were 0.03, 1, 2, and 4 microgram/ml, respectively. Overall rates of resistance to trovafloxacin (0.3%), levofloxacin (0.3%), grepafloxacin (0.6%), and ofloxacin (0.7%) were low. For ofloxacin-intermediate and -resistant isolates (n = 142), gemifloxacin had the lowest MIC(90) (0.12 microgram/ml) compared to the MIC(90)s of trovafloxacin (0.5 microgram/ml), grepafloxacin (1 microgram/ml), and levofloxacin (2 microgram/ml). For all S. pneumoniae isolates tested, gemifloxacin MICs were 2 log(10) CFU, compared to those in untreated controls, for both vancomycin susceptible and -resistant (VanA and VanB) E. faecalis strains and >4 log(10) CFU for a methicillin-resistant S. aureus isolate. The glycylcycline was more efficacious at a lower administered dose in the rat model of endocarditis than was vancomycin. The efficacy of GAR-936 in this model was apparently not enhanced by a factor in rat serum, as was observed for vancomycin with a time-kill curve. The results of this study demonstrate the therapeutic potential of GAR-936 for the treatment of enterococcal and staphylococcal infections and warrant further investigation. PMID- 11036016 TI - Activity of LY333328 combined with gentamicin in vitro and in rabbit experimental endocarditis due to vancomycin-susceptible or -resistant Enterococcus faecalis. AB - We investigated the activity of LY333328 alone and combined with gentamicin, both in vitro and in a rabbit model of experimental endocarditis, against the susceptible strain Enterococcus faecalis JH2-2 and its two glycopeptide-resistant transconjugants, BM4316 (VanA) and BM4275 (VanB). MICs of LY333328 and gentamicin were 2 and 16 microgram/ml, respectively, for the three strains. In vitro, LY333328 alone was bactericidal at 24 h against JH2-2 at a concentration of 2 microgram/ml and against BM4316 and BM4275 at a concentration of 30 microgram/ml. The combination of LY333328 and gentamicin (4 microgram/ml) was synergistic and bactericidal after 24 h of incubation against the three strains at LY333328 concentrations of 2 microgram/ml for JH2-2 and 8 microgram/ml for BM4275 and BM4316. The combination of LY333328 and gentamicin was the only regimen demonstrating in vitro bactericidal activity against BM4316. In vivo, intravenous treatment with LY333328 alone, providing peak and trough serum levels of 83.3 +/- 1.3 and 3.8 +/- 0.2 microgram/ml, respectively, was inactive against BM4316 and BM4275 and selected mutants resistant to LY333328 in half of the rabbits infected with the VanA-type strain (MICs, 8 to 20 microgram/ml). However, the LY333328 gentamicin combination was active against the three strains and prevented the emergence of mutants resistant to both components of the combination. We conclude that the LY333328-gentamicin combination might be of interest for the treatment of enterococcal infections, particularly against VanA-type strains. PMID- 11036018 TI - Genetic diversity of carbapenem-hydrolyzing metallo-beta-lactamases from Chryseobacterium (Flavobacterium) indologenes. AB - The class B carbapenem-hydrolyzing beta-lactamase IND-1 has been characterized for Chryseobacterium indologenes strain 001. With internal primers for the bla gene for IND-1 (bla(IND-1)) and an internal bla(IND-1) probe, PCR amplifications failed, while hybridization results were positive when DNA from another C. indologenes isolate, strain CIP101026, was used as a template. Thus, a bla(IND) related gene was cloned from this C. indologenes reference strain. Sequencing of the insert of a recombinant plasmid conferring resistance to carbapenems revealed an open reading frame with a G + C content of 39.9% and coding for a 243-amino acid preprotein named IND-2. IND-2 shared 80% amino acid identity with IND-1 and had a similar broad-spectrum resistance profile, including resistance to carbapenems. It was classified in functional subgroup 3a of class B carbapenem hydrolyzing beta-lactamases. IND-1 and IND-2, despite their genetic diversity, possessed similar kinetic parameters, except that ceftazidime was hydrolyzed less by IND-2. To obtain the entire bla(IND)-related gene sequences of eight other C. indologenes isolates, PCR was performed using internal and external primers, followed by inverse PCR techniques. The likely chromosome-mediated metallo-beta lactamases of the 10 C. indologenes isolates were divided into several groups and subgroups. IND-1, IND-2, IND-2a, IND-3, and IND-4 shared 77 to 99% amino acid identity. PMID- 11036019 TI - SME-type carbapenem-hydrolyzing class A beta-lactamases from geographically diverse Serratia marcescens strains. AB - Three sets of carbapenem-resistant Serratia marcescens isolates have been identified in the United States: 1 isolate in Minnesota in 1985 (before approval of carbapenems for clinical use), 5 isolates in Los Angeles (University of California at Los Angeles [UCLA]) in 1992, and 19 isolates in Boston from 1994 to 1999. All isolates tested produced two beta-lactamases, an AmpC-type enzyme with pI values of 8.6 to 9.0 and one with a pI value of approximately 9.5. The enzyme with the higher pI in each strain hydrolyzed carbapenems and was not inhibited by EDTA, similar to the chromosomal class A SME-1 beta-lactamase isolated from the 1982 London strain S. marcescens S6. The genes encoding the carbapenem hydrolyzing enzymes were cloned in Escherichia coli and sequenced. The enzyme from the Minnesota isolate had an amino acid sequence identical to that of SME-1. The isolates from Boston and UCLA produced SME-2, an enzyme with a single amino acid change relative to SME-1, a substitution from valine to glutamine at position 207. Purified SME enzymes from the U. S. isolates had beta-lactam hydrolysis profiles similar to that of the London SME-1 enzyme. Pulsed-field gel electrophoresis analysis revealed that the isolates showed some similarity but differed by at least three genetic events. In conclusion, a family of rare class A carbapenem-hydrolyzing beta-lactamases first described in London has now been identified in S. marcescens isolates across the United States. PMID- 11036020 TI - Identification of the coumermycin A(1) biosynthetic gene cluster of Streptomyces rishiriensis DSM 40489. AB - The biosynthetic gene cluster of the aminocoumarin antibiotic coumermycin A(1) was cloned by screening of a cosmid library of Streptomyces rishiriensis DSM 40489 with heterologous probes from a dTDP-glucose 4,6-dehydratase gene, involved in deoxysugar biosynthesis, and from the aminocoumarin resistance gyrase gene gyrB(r). Sequence analysis of a 30.8-kb region upstream of gyrB(r) revealed the presence of 28 complete open reading frames (ORFs). Fifteen of the identified ORFs showed, on average, 84% identity to corresponding ORFs in the biosynthetic gene cluster of novobiocin, another aminocoumarin antibiotic. Possible functions of 17 ORFs in the biosynthesis of coumermycin A(1) could be assigned by comparison with sequences in GenBank. Experimental proof for the function of the identified gene cluster was provided by an insertional gene inactivation experiment, which resulted in an abolishment of coumermycin A(1) production. PMID- 11036021 TI - Fluoroquinolone resistance in clinical isolates of Streptococcus pneumoniae: contributions of type II topoisomerase mutations and efflux to levels of resistance. AB - We report on amino acid substitutions in the quinolone resistance-determining region of type II topisomerases and the prevalence of reserpine-inhibited efflux for 70 clinical isolates of S. pneumoniae for which the ciprofloxacin MIC is >/=4 microgram/ml and 28 isolates for which the ciprofloxacin MIC is 64 microgram/ml for RPZ-TH, RPZ, lansoprazole, and omeprazole, respectively. No such inhibitory effects were observed with H(2) blockers or anti H. pylori agents. The motilities of Campylobacter jejuni and C. coli-but not those of Vibrio cholerae O1 and O139, Vibrio parahaemolyticus, Salmonella enterica serovar Typhimurium, and Proteus mirabilis-were also inhibited. Prolonged incubation with RPZ or RPZ-TH inhibited bacterial growth of only H. pylori, except for a turbid colony mutant. The results indicate that RPZ and RPZ TH have a characteristic inhibitory effect against the motility of H. pylori (spiral-shaped bacteria), which is distinguished from that against bacterial growth. PMID- 11036025 TI - Flow cytometric detection of Leishmania parasites in human monocyte-derived macrophages: application to antileishmanial-drug testing. AB - A flow cytometric technique was developed for detection of amastigotes of the protozoan Leishmania infantum in human nonadherent monocyte-derived macrophages. The cells were fixed and permeabilized with paraformaldehyde-ethanol, and intracellular amastigotes were labeled with Leishmania lipophosphoglycan-specific monoclonal antibody. Results showed that flow cytometry provided accurate quantification of the infection rates in human macrophages compared to the rates obtained by the conventional microscopic technique, with the advantage that a large number of cells could be analyzed rapidly. The results demonstrated, moreover, that labeling of intracellular amastigotes could reliably be used to evaluate the antileishmanial activities of conventional drugs such as meglumine antimoniate, amphotericin B, pentamidine, and allopurinol. They also established that various Leishmania species (L. mexicana, L. donovani) could be detected by this technique in other host-cell models such as mouse peritoneal macrophages and suggested that the flow cytometric method could be a valid alternative to the conventional method. PMID- 11036026 TI - Cloning and characterization of SmeDEF, a novel multidrug efflux pump from Stenotrophomonas maltophilia. AB - Stenotrophomonas maltophilia is a nosocomial bacterial pathogen intrinsically resistant to several antibiotics. The mechanisms involved in this intrinsic multiresistance phenotype are poorly understood. A library of chromosomal DNA from a spontaneous multidrug-resistant S. maltophilia D457R mutant (A. Alonso and J. L. Martinez, Antimicrob. Agents Chemother. 41:1140-1142, 1997) was screened for complementation of erythromycin susceptibility on an antibiotic hypersusceptible Escherichia coli DeltaacrAB strain. Cloning and further analysis revealed that a 6-kbp region constituting a transcriptional unit was capable of complementing the antibiotic-susceptible phenotype of an E. coli DeltaacrAB strain. We identified three open reading frames, smeD, smeE and smeF, which code for members of the membrane fusion protein, resistance nodulation division, and outer membrane factor families, respectively. Drug susceptibility assays indicated that the SmeDEF system cloned in E. coli mediates resistance to a wide range of antibiotics. Ethidium bromide and norfloxacin accumulation experiments in the presence and in the absence of carbonyl cyanide m-chlorophenylhydrazone showed that this system constitutes a drug efflux pump dependent on the membrane proton motive force. The presence of high levels of smeDEF mRNA in the multiresistant D457R mutant was consistent with the high levels of SmeF (formerly Omp54) observed in the same strain. In contrast, transcription levels of smeDEF in the D457 strain were tiny, which correlates with the low levels of SmeF observed for this strain. Also, for both the D457 and D457R strains, we observed growth phase-dependent regulation in which the highest level of transcription corresponded to early exponential phase, with transcription decreasing throughout the growth curve to undetectable levels at 24 h. PMID- 11036027 TI - Comparative efficacies of terbinafine and fluconazole in treatment of experimental coccidioidal meningitis in a rabbit model. AB - A rabbit model of coccidioidal meningitis was used to compare the therapeutic efficacies of terbinafine (TBF) and fluconazole (FCZ). Hydrocortisone acetate treated New Zealand White male rabbits were infected intracisternally with either 2.2 x 10(4) or 6.4 x 10(4) Coccidioides immitis arthroconidia. Oral treatment with polyethylene glycol 200 (PEG) twice daily (n = 8), TBF twice daily (n = 9; 200 mg/kg of body weight/day), or FCZ once daily (n = 8; 80 mg/kg/day) began on day 5 and continued for 21 days. Mean survival times were 20, 24, and 32 days for rabbits treated with PEG, TBF, and FCZ, respectively. All of the FCZ-treated animals (100%; P = 0.003), 56% of the TBF-treated animals (P = 0.4), and 25% of the PEG-treated animals survived the length of the study. Both FCZ and TBF were effective at reducing the incidence of paresis. Only FCZ was effective at reducing most neurological and systemic signs. FCZ treatments resulted in lower cerebrospinal fluid (CSF) protein concentrations and leukocyte counts and faster clearing of CSF fungal cultures compared with those for PEG-treated controls, but TBF treatments had no significant effect on these parameters. Neither drug affected CSF glucose levels. Mean serum TBF levels by bioassay were within the range of 3.5 to 6.2 microgram/ml at 1, 2, and 4 h postdosing and 0.35 to 7.0 microgram/ml at 14 h postdosing. No TBF was detected in CSF. Mean FCZ levels (24 to 25.5 h postdosing) by bioassay were 16.4 to 19.2 and 13.5 to 19.2 microgram/ml in serum and CSF, respectively. The reduction in the numbers of CFU in the spinal cord and brain was over 100-fold (P = 0.0005) in FCZ-treated animals and 2-fold (P C), 2480 (C --> T), 2535 (G --> A), and 2536 (G --> C). The mutations map within two different stems of the peptidyltransferase region of domain V. Because multiple copies of rDNA are present in the chromosome, gene conversion between mutant and wild-type 23S rDNA alleles may be necessary for stable resistance. Additionally, none of the characterized mutants showed cross resistance to any of a spectrum of protein synthesis inhibitors, suggesting that the target site of evernimicin may be unique. PMID- 11036031 TI - Gametocytocidal activity and synergistic interactions of riboflavin with standard antimalarial drugs against growth of Plasmodium falciparum in vitro. AB - Our previous studies have shown that riboflavin has activity against Plasmodium falciparum asexual-stage parasites in vitro. In the present study we examine the gametocytocidal activity of riboflavin and the interaction of riboflavin with some commonly used antimalarial drugs against the asexual forms of P. falciparum in vitro. The addition of riboflavin to P. falciparum cultures killed gametocytes at all stages, even those at late stages (III to V), which are not affected by many of the commonly used antimalarials. Combinations of riboflavin with mefloquine, pyrimethamine, and quinine showed a marked potentiation of the activities of these drugs against asexual-stage parasites in vitro. The combination of riboflavin with artemisinin was additive, while that with chloroquine was mildly antagonistic. High doses of riboflavin are used clinically to treat several inborn errors of metabolism with no adverse side effects. Its efficacy in combination with standard antimalarial drugs in treating and preventing the transmission of P. falciparum malaria can therefore be evaluated in humans. PMID- 11036032 TI - Potent antipneumococcal activity of gemifloxacin is associated with dual targeting of gyrase and topoisomerase IV, an in vivo target preference for gyrase, and enhanced stabilization of cleavable complexes in vitro. AB - We investigated the roles of DNA gyrase and topoisomerase IV in determining the susceptibility of Streptococcus pneumoniae to gemifloxacin, a novel fluoroquinolone which is under development as an antipneumococcal drug. Gemifloxacin displayed potent activity against S. pneumoniae 7785 (MIC, 0.06 microgram/ml) compared with ciprofloxacin (MIC, 1 to 2 microgram/ml). Complementary genetic and biochemical approaches revealed the following. (i) The gemifloxacin MICs for isogenic 7785 mutants bearing either parC or gyrA quinolone resistance mutations were marginally higher than wild type at 0.12 to 0.25 microgram/ml, whereas the presence of both mutations increased the MIC to 0.5 to 1 microgram/ml. These data suggest that both gyrase and topoisomerase IV contribute significantly as gemifloxacin targets in vivo. (ii) Gemifloxacin selected first-step gyrA mutants of S. pneumoniae 7785 (gemifloxacin MICs, 0.25 microgram/ml) encoding Ser-81 to Phe or Tyr, or Glu-85 to Lys mutations. These mutants were cross resistant to sparfloxacin (which targets gyrase) but not to ciprofloxacin (which targets topoisomerase IV). Second-step mutants (gemifloxacin MICs, 1 microgram/ml) exhibited an alteration in parC resulting in changes of ParC hot spot Ser-79 to Phe or Tyr. Thus, gyrase appears to be the preferential in vivo target. (iii) Gemifloxacin was at least 10- to 20-fold more effective than ciprofloxacin in stabilizing a cleavable complex (the cytotoxic lesion) with either S. pneumoniae gyrase or topoisomerase IV enzyme in vitro. These data suggest that gemifloxacin is an enhanced affinity fluoroquinolone that acts against gyrase and topoisomerase IV in S. pneumoniae, with gyrase the preferred in vivo target. The marked potency of gemifloxacin against wild type and quinolone-resistant mutants may accrue from greater stabilization of cleavable complexes with the target enzymes. PMID- 11036033 TI - Evidence for an efflux pump mediating multiple antibiotic resistance in Salmonella enterica serovar Typhimurium. AB - The mechanism of multiple antibiotic resistance in six isolates of Salmonella enterica serovar Typhimurium recovered from a patient treated with ciprofloxacin was studied to investigate the role of efflux in the resistance phenotype. Compared to the patient's pretherapy isolate (L3), five of six isolates accumulated less ciprofloxacin, three of six isolates accumulated less chloramphenicol, and all six accumulated less tetracycline. The accumulation of one or more antibiotics was increased by carbonyl cyanide m-chlorophenylhydrazone to concentrations similar to those accumulated by L3 for all isolates except one, in which accumulation of all three agents remained approximately half that of L3. All isolates had the published wild-type sequences of marO and marR. No increased expression of marA, tolC, or soxS was observed by Northern blotting; however, three isolates showed increased expression of acrB, which was confirmed by quantitative competitive reverse transcription-PCR. However, there were no mutations within acrR or the promoter region of acrAB in any of the isolates. PMID- 11036034 TI - Glycopeptide susceptibility profiles of Staphylococcus haemolyticus bloodstream isolates. AB - Twelve clinical strains of Staphylococcus haemolyticus (eight methicillin resistant and three methicillin susceptible), isolated from blood cultures between 1982 and 1997, were investigated for teicoplanin and vancomycin susceptibility profiles. On the basis of conventional MIC tests and breakpoints, four isolates were susceptible (MICs, 1 to 8 microgram/ml) and eight were resistant (MICs, 32 to 64 microgram/ml) to teicoplanin while all were susceptible to vancomycin (MICs, 1 to 2 microgram/ml). All four strains for which the conventional teicoplanin MICs were within the range of susceptibility expressed heterogeneous resistance to teicoplanin and homogeneous vancomycin susceptibility. Of the eight strains for which the conventional teicoplanin MICs were within the range of resistance, six expressed heterogeneous and two expressed homogeneous teicoplanin resistance while seven showed heterogeneous vancomycin resistance profiles (with subpopulations growing on 8 microgram of the drug per ml at frequencies of >/=10(-6) for six strains and 10(-7) for one) and one demonstrated homogeneous vancomycin susceptibility. Of six bloodstream isolates of other staphylococcal species (S. aureus, S. epidermidis, and S. simulans), for all of which the conventional teicoplanin MICs were >/=4 microgram/ml and the vancomycin MICs were 8-fold-reduced sensitivity, respectively). The mutations 46I/L, 54L/V, 84V, and 90M showed the strongest association with amprenavir resistance (P < 0. 0001). The scoring system using 84V and/or any two of a number of mutations (10I/R/V/F, 46I/L, 54L/V, and 90M) predicted amprenavir resistance with a sensitivity of 86.0% and a specificity of 81.0% within the analyzed group of samples. Of 62 samples with resistance against 4 PI, 23 (37.1%) were still sensitive to amprenavir. In comparison, only 2 of 23 samples (8.7%) from nelfinavir-naive patients with resistance against indinavir, saquinavir, and ritonavir were still sensitive to nelfinavir. Amprenavir thus appears to be an interesting alternative for PI salvage therapy. PMID- 11036058 TI - Effect of ceftazidime on systemic cytokine concentrations in rats. AB - The effect of a single dose of ceftazidime on circulating concentrations of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) in a rat model of sepsis was studied. IL-6 concentrations were significantly elevated (100 to 200 times the baseline) 6 h after ceftazidime administration in both septic and nonseptic (control) rats. TNF-alpha concentrations increased significantly in nonseptic (approximately 40 times the baseline) rats but not septic (approximately 2 to 3 times the baseline) rats. Ceftazidime administration was not associated with an increase in endotoxin concentrations. These findings suggest that ceftazidime modulation of proinflammatory cytokine concentrations may be independent of its antimicrobial properties. PMID- 11036059 TI - Heterogeneity of AmpC cephalosporinases of Hafnia alvei clinical isolates expressing inducible or constitutive ceftazidime resistance phenotypes. AB - Ten unrelated Hafnia alvei clinical isolates were grouped according to either their low-level and inducible cephalosporinase production or their high-level and constitutive cephalosporinase production phenotype. Their AmpC sequences shared 85 to 100% amino acid identity. The immediate genetic environment of ampC genes was conserved in H. alvei isolates but was different from that found in other ampC-possessing enterobacterial species. PMID- 11036060 TI - Genetic characterization of vanG, a novel vancomycin resistance locus of Enterococcus faecalis. AB - Enterococcus faecalis strain WCH9 displays a moderate level of resistance to vancomycin (MIC = 16 microgram/ml) and full susceptibility to teicoplanin but is negative by PCR analysis using primers specific for all known enterococcal vancomycin resistance genotypes (vanA, vanB, vanC, vanD, and vanE). We have isolated and sequenced a novel putative vancomycin resistance locus (designated vanG), which contains seven open reading frames, from this strain. These are organized differently from those of all the other enterococcal van loci, and, furthermore, the individual vanG gene products exhibit less than 50% amino acid sequence identity to other van gene products. PMID- 11036061 TI - Development of resistance to ciprofloxacin, rifampin, and mupirocin in methicillin-susceptible and -resistant Staphylococcus aureus isolates. AB - A relationship between resistance to methicillin and resistance to fluoroquinolones, rifampin, and mupirocin has been described for Staphylococcus aureus. Differences in resistance rates may be explainable by a higher spontaneous mutation rate (MR) or a faster development of resistance (DIFF) in methicillin-resistant S. aureus (MRSA). No differences in MR, DIFF, and mutations in grlA and gyrA were detected between methicillin-susceptible S. aureus and MRSA. The higher resistance rates in MRSA are not the result of hypermutability of target genes or a faster emergence of different mutations and may be the consequence of clonal spread of multiresistant MRSA. PMID- 11036062 TI - Updated sequence information and proposed nomenclature for bla(TEM) genes and their promoters. AB - The nucleotide sequences of 59 bla(TEM) genes encoding inhibitor-resistant TEM enzymes showed great genetic variability and were associated with different types of promoters. These findings led us to suggest an updated bla(TEM) gene nomenclature based on the origin of the bla(TEM) gene (bla(TEM-1A), bla(TEM-1B), bla(TEM-1C), bla(TEM-1D), bla(TEM-1E), and bla(TEM-1F)) and the promoter type. PMID- 11036063 TI - Suppression of posttreatment recurrence of experimental visceral Leishmaniasis in T-cell-deficient mice by oral miltefosine. AB - T-cell-deficient nude mice infected with Leishmania donovani were treated with miltefosine and then given either no treatment or intermittent miltefosine. Intracellular visceral infection recurred in untreated mice but was suppressed by once- or twice-weekly oral administration of miltefosine. Miltefosine may be useful as oral maintenance therapy for T-cell-deficient patients with visceral leishmaniasis. PMID- 11036064 TI - Mints as adaptors. Direct binding to neurexins and recruitment of munc18. AB - Mint1 (X11/human Lin-10) and Mint2 are neuronal adaptor proteins that bind to Munc18-1 (n/rb-sec1), a protein essential for synaptic vesicle exocytosis. Mint1 has previously been characterized in a complex with CASK, another adaptor protein that in turn interacts with neurexins. Neurexins are neuron-specific cell surface proteins that act as receptors for the excitatory neurotoxin alpha-latrotoxin. Hence, one possible function for Mint1 is to mediate the recruitment of Munc18 to neurexins. In agreement with this hypothesis, we now show that the cytoplasmic tail of neurexins captures Munc18 via a multiprotein complex that involves Mint1. Furthermore, we demonstrate that both Mint1 and Mint2 can directly bind to neurexins in a PDZ domain-mediated interaction. Various Mint and/or CASK containing complexes can be assembled on neurexins, and we demonstrate that Mint1 can bind to Munc18 and CASK simultaneously. Our data support a model whereby one of the functions of Mints is to localize the vesicle fusion protein Munc18 to those sites at the plasma membrane that are defined by neurexins, presumably in the vicinity of points of exocytosis. PMID- 11036065 TI - The yeast Na+/H+ exchanger Nhx1 is an N-linked glycoprotein. Topological implications. AB - Nhx1, the endosomal Na(+)/H(+) exchanger of Saccharomyces cerevisiae represents the founding member of a newly emerging subfamily of intracellular Na(+)/H(+) exchangers. These proteins share significantly greater sequence homology to one another than to members of the mammalian Na(+)/H(+) exchanger (NHE) family encoding plasma membrane Na(+)/H(+) exchangers. Members of both subtypes are predicted to share a common organization, with an N-terminal transporter domain of transmembrane helices followed by a C-terminal hydrophilic tail. In the present study, we show that Nhx1 is an asparagine-linked glycoprotein and that the sites of glycosylation map to two residues within the C-terminal stretch of the polypeptide. This is the first evidence, to date, for glycosylation of the C terminal region of any known NHE isoform. Importantly, the mapping of N-linked glycosylation to the C-terminal domain of Nhx1 is indicative of an unexpected membrane topology, particularly with regard to the orientation of the tail region. Although one recent study demonstrated that certain epitopes in the C terminal domain of NHE3 were accessible from the exoplasmic side of the plasma membrane (Biemesderfer, D., DeGray, B., and Aronson, P. S. (1998) J. Biol. Chem. 273, 12391-12396), numerous other studies implicate a cytosolic disposition for the hydrophilic C-terminal tail of plasma membrane NHE isoforms. Our analysis of the glycosylation of Nhx1 is strongly indicative of residence of at least some portion of the hydrophilic tail domain within the endosomal lumen. These findings imply that the organization of the tail domain may be more complex than previously assumed. PMID- 11036066 TI - The role of the C1 and C2 a-domains in type VI collagen assembly. AB - Constructs of each of the three chains of type VI collagen were generated and examined in an in vitro transcription/translation assay supplemented with semipermeabilized cells. Each of the constructs when used in the in vitro system was shown to be glycosylated and to undergo intracellular assembly, the extent of which was determined by the nature of the C-terminal globular domains. All three chains containing the C1 domain formed monomers; however, the C2 domain was required for dimer and tetramer formation. In the case of the full-length alpha2(VI) chain, monomers, dimers, and tetramers formed in a time-dependent manner. Although the splice variant alpha2(VI)C2a could form monomers, it was unable to form dimers and tetramers. Similar results to the alpha2(VI) chain were found for the full-length alpha1(VI) chain, although assembly was at a slower rate. In the case of the alpha3(VI) chain containing both C1 and C2 domains only monomers were observed. Addition of the C3, C4, and C5 did not change this pattern. Homology modeling suggested that a 10-amino acid insertion in the C2 domain of the alpha3(VI) chain may interfere with dimer formation. A near full length construct of the alpha3(VI) chain only formed monomers but was shown to facilitate tetramer formation in cotranslation experiments. PMID- 11036067 TI - In vitro binding of ribosomes to the beta subunit of the Sec61p protein translocation complex. AB - The Sec61p complex forms the core element of the protein translocation complex (translocon) in the rough endoplasmic reticulum (rough ER) membrane. Translating or nontranslating ribosomes bind with high affinity to ER membranes that have been stripped of ribosomes or to liposomes containing purified Sec61p. Here we present evidence that the beta subunit of the complex (Sec61beta) makes contact with nontranslating ribosomes. A fusion protein containing the Sec61beta cytoplasmic domain (Sec61beta(c)) prevents the binding of ribosomes to stripped ER-derived membranes and also binds to ribosomes directly with an affinity close to the affinity of ribosomes for stripped ER-derived membranes. The ribosome binding activity of Sec61beta(c), like that of native ER membranes, is sensitive to high salt concentrations and is not based on an unspecific charge-dependent interaction of the relatively basic Sec61beta(c) domain with ribosomal RNA. Like stripped ER membranes, the Sec61beta(c) sequence binds to large ribosomal subunits in preference over small subunits. Previous studies have shown that Sec61beta is inessential for ribosome binding and protein translocation, but translocation is impaired by the absence of Sec61beta, and it has been proposed that Sec61beta assists in the insertion of nascent proteins into the translocation pore. Our results suggest a physical interaction of the ribosome itself with Sec61beta; this may normally occur alongside interactions between the ribosome and other elements of Sec61p, or it may represent one stage in a temporal sequence of binding. PMID- 11036068 TI - Acceleration of thrombomodulin gene transcription by retinoic acid: retinoic acid receptors and Sp1 regulate the promoter activity through interactions with two different sequences in the 5'-flanking region of human gene. AB - The interactions between retinoic acid- (RA)-dependent transcriptional regulatory sequences of the 5'-untranslated region of the thrombomodulin gene and nuclear RA responsive proteins were studied using human pancreas BxPC-3 cells. Deletion mutants of pTM-CAT plasmid revealed the presence of distal and proximal RA responsive regions containing direct repeat with 4 spaces (DR4) and three of four Sp1 sites, respectively. Cotransfection of a pTM-CAT plasmid with expression plasmids of RA receptors (RARalpha, RARbeta, and RARgamma) augmented the promoter activity under the condition of lower retinoid X receptor-alpha (RXRalpha) expression, whereas the activity was greatly diminished when RXRalpha was highly expressed. An electrophoretic mobility shift assay with cDNA containing the DR4 indicated that heterodimers of RAR and RXRalpha interacted with the DR4 site, although the interaction gradually disappeared with the increase in the ratio of RXRalpha/RAR. On the other hand, Sp1 protein interacted especially with the tandem Sp1 site corresponding to the first and second Sp1 sequences of the four Sp1 sites in the proximal RA-responsive region. The binding of Sp1 to Sp1 sites was independent of RAR-RXR heterodimer but increased with the increase in Sp1 concentration in the presence of unknown factor(s) of reticulocyte lysate. Upon treatment of the cells with RA, time-dependent increases in the ratio of RARbeta to RXRalpha and the phosphorylated form of Sp1 were observed. We concluded that two genomic DNA regions, the DR4 site (-1531 to -1516) and the first and second Sp1-binding sites (-145 to -121), were involved in the RA-dependent augmentation of thrombomodulin gene expression through increased interactions of the two regions with heterodimer of RAR-RXRalpha and nuclear Sp1, respectively. PMID- 11036069 TI - The M3 muscarinic acetylcholine receptor expressed in HEK-293 cells signals to phospholipase D via G12 but not Gq-type G proteins: regulators of G proteins as tools to dissect pertussis toxin-resistant G proteins in receptor-effector coupling. AB - The M(3) muscarinic acetylcholine receptor (mAChR) expressed in HEK-293 cells couples to G(q) and G(12) proteins and stimulates phospholipase C (PLC) and phospholipase D (PLD) in a pertussis toxin-insensitive manner. To determine the type of G protein mediating M(3) mAChR-PLD coupling in comparison to M(3) mAChR PLC coupling, we expressed various Galpha proteins and regulators of the G protein signaling (RGS), which act as GTPase-activating proteins for G(q)- or G(12)-type G proteins. PLD stimulation by the M(3) mAChR was enhanced by the overexpression of Galpha(12) and Galpha(13), whereas the overexpression of Galpha(q) strongly increased PLC activity without affecting PLD activity. Expression of the RGS homology domain of Lsc, which acts specifically on Galpha(12) and Galpha(13), blunted the M(3) mAChR-induced PLD stimulation without affecting PLC stimulation. On the other hand, overexpression of RGS4, which acts on Galpha(q)- but not Galpha(12)-type G proteins, suppressed the M(3) mAChR induced PLC stimulation without altering PLD stimulation. We conclude that the M(3) mAChR in HEK-293 cells apparently signals to PLD via G(12)- but not G(q) type G proteins and that G protein subtype-selective RGS proteins can be used as powerful tools to dissect the pertussis toxin-resistant G proteins and their role in receptor-effector coupling. PMID- 11036070 TI - The crystal structure of the fab fragment of the monoclonal antibody MAK33. Implications for folding and interaction with the chaperone bip. AB - The Fab fragment of the murine monoclonal antibody, MAK33, directed against human creatine kinase of the muscle-type, was crystallized and the three-dimensional structure was determined to 2.9 A. The antigen-binding surface of MAK33 shows a convex overall shape typical for immunoglobulins binding large antigens. The structure allows us to analyze the environment of cis-prolyl-peptide bonds whose isomerization is of key importance in the folding process. These residues seem to be involved with not only domain stability but also seem to play a role in the association of heavy and light chains, reinforcing the importance of beta-strand recognition in antibody assembly. The structure also allows the localization of segments of primary sequence postulated to represent binding sites for the ER specific chaperone BiP within the context of the entire Fab fragment. These sequences are found primarily in beta-strands that are necessary for interactions between the individual domains. PMID- 11036071 TI - Human T-lymphotropic virus type I Tax protein utilizes distinct pathways for p53 inhibition that are cell type-dependent. AB - p53 plays a pivotal role in transmitting signals from many forms of genotoxic stress to genes and factors that control the cell cycle and apoptosis. We have previously shown that the human T-lymphotropic virus type I Tax protein can inhibit p53 function. Recently we reported that Tax inhibits p53 function in Jurkat cells and mouse embryo fibroblasts through a mechanism involving the nuclear factor kappa B pathway and correlates with phosphorylation on serines 15 and 392 of p53. However, several groups have also observed a mechanism that correlates with p300 binding of Tax. To address this controversy and to determine the mechanism by which Tax inhibits p53 function, we examined the activation functions of Tax required for p53 inhibition. In HeLa and H1299 cells the cAMP response element-binding protein/activating transcription factor activation function is essential, as demonstrated by the Tax mutants M47 and K88A. In addition, expression of exogenous p300 in H1299 cells allows full recovery of p53 transactivation in the presence of Tax. Consistent with p300 being a limiting factor in H1299, Saos-2, and HeLa cells, we found that the level of endogenous p300 is relatively low in these cells compared with Jurkat cells or the human T lymphotropic virus type I-infected C81 and MT2 cells. Thus our data suggests that Tax utilizes distinct mechanisms to inhibit p53 function that are cell type dependent. PMID- 11036072 TI - Mitochondrial calcium oscillations in C2C12 myotubes. AB - Mitochondrial Ca(2+) concentration ([Ca(2+)](m)) was monitored in C2C12 skeletal muscle cells stably expressing the Ca(2+)-sensitive photoprotein aequorin targeted to mitochondria. In myotubes, KCl-induced depolarization caused a peak of 3.03 +/- 0.14 micrometer [Ca(2+)](m) followed by an oscillatory second phase (5.1 +/- 0.1 per min). Chelation of extracellular Ca(2+) or blockade of the voltage-operated Ca(2+) channel attenuated both phases of the KCl response. The inhibitor of the sarcoplasmic reticulum Ca(2+)-ATPase, cyclopiazonic acid, reduced the amplitude of the KCl-induced [Ca(2+)](m) peak and prevented the oscillations, suggesting that these were generated intracellularly. No such [Ca(2+)](m) oscillations occurred with the nicotinic agonist carbachol, cyclopiazonic acid alone, or the purinergic agonist ATP. In contrast, caffeine produced an oscillatory behavior, indicating a role of ryanodine receptors as mediators of the oscillations. The [Ca(2+)](m) response was desensitized when cells were exposed to two consecutive challenges with KCl separated by a 5-min wash, whereas a second pulse of carbachol potentiated [Ca(2+)](m), indicating differences in intracellular Ca(2+) redistribution. Cross-desensitization between KCl and carbachol and cross-potentiation between carbachol and KCl were observed. These results suggest that close contacts between mitochondria and sarcoplasmic reticulum exist permitting Ca(2+) exchanges during KCl depolarization. These newly demonstrated dynamic changes in [Ca(2+)](m) in stimulated skeletal muscle cells might contribute to the understanding of physiological and pathological processes in muscular disorders. PMID- 11036073 TI - ESE-1 is a novel transcriptional mediator of inflammation that interacts with NF kappa B to regulate the inducible nitric-oxide synthase gene. AB - Inflammation is a hallmark of several vascular diseases. The nuclear factor kappaB (NF-kappaB) transcription factors are dimeric proteins involved in the activation of a large number of genes in response to inflammatory stimuli. We report the involvement of a novel member of the ETS transcription factor, ESE-1, in mediating vascular inflammation. ESE-1 is induced in response to inflammatory cytokines and lipopolysaccharide in vascular smooth muscle cells, endothelial cells, and cells of the monocyte-macrophage lineage. This induction occurs within hours of stimulation and is mediated by NF-kappaB transactivation of the ESE-1 promoter. We have identified the inducible form of nitric-oxide synthase (NOS2) as a putative target for ESE-1. ESE-1 can bind to the p50 subunit of NF-kappaB, and cotransfection of ESE-1 with the p50 and p65 subunits of NF-kappaB synergistically enhances transactivation of the NOS2 promoter by ESE-1. An ESE-1 binding site within the NOS2 promoter has been identified, the site-directed mutagenesis of which completely abolishes the ability of ESE-1 to transactivate the NOS2 promoter. Finally, in a mouse model of endotoxemia, associated with acute vascular inflammation, ESE-1 is strongly expressed in vascular endothelium and smooth muscle cells. In summary, ESE-1 represents a novel mediator of vascular inflammation. PMID- 11036074 TI - Sequence-specific and methylation-dependent and -independent binding of rice nuclear proteins to a rice tungro bacilliform virus vascular bundle expression element. AB - Nuclear proteins from rice (Oryza sativa) were identified that bind specifically to a rice tungro bacilliform virus promoter region containing a vascular bundle expression element (VBE). One set of proteins of 29, 33, and 37 kDa, present in shoot and cell suspension extracts but hardly detectable in root extracts, bound to a site containing the sequence AGAAGGACCAGA within the VBE, which also contains two CpG and one CpNpG potential methylation motifs. Binding by these proteins was determined to be cytosine methylation-independent. However, a novel protein present in all analyzed extracts bound specifically to the methylated VBE. A region of at least 49 nucleotides overlapping the VBE and complete cytosine methylation of the three Cp(Np)G motifs was required for efficient binding of this methylated VBE-binding protein (MVBP). PMID- 11036075 TI - The role of N-glycosylation in transport to the plasma membrane and sorting of the neuronal glycine transporter GLYT2. AB - Glycine transporter GLYT2 is an axonal glycoprotein involved in the removal of glycine from the synaptic cleft. To elucidate the role of the carbohydrate moiety on GLYT2 function, we analyzed the effect of the disruption of the putative N glycosylation sites on the transport activity, intracellular traffic in COS cells, and asymmetrical distribution of this protein in polarized Madin-Darby canine kidney (MDCK) cells. Transport activity was reduced by 35-40% after enzymatic deglycosylation of the transporter reconstituted into liposomes. Site directed mutagenesis of the four glycosylation sites (Asn-345, Asn-355, Asn-360, and Asn-366), located in the large extracellular loop of GLYT2, produced an inactive protein that was retained in intracellular compartments when transiently transfected in COS cells or in nonpolarized MDCK cells. When expressed in polarized MDCK cells, wild type GLYT2 localizes in the apical surface as assessed by transport and biotinylation assays. However, a partially unglycosylated mutant (triple mutant) was distributed in a nonpolarized manner in MDCK cells. The apical localization of GLYT2 occurred by a glycolipid rafts independent pathway. PMID- 11036076 TI - Interaction kinetics of reversible inhibitors and substrates with acetylcholinesterase and its fasciculin 2 complex. AB - Fasciculin 2 (Fas2), a three-fingered peptide of 61 amino acids, binds tightly to the peripheral site of acetylcholinesterases (AChE; EC ), occluding the entry portal into the active center gorge of the enzyme and inhibiting its catalytic activity. We investigated the mechanism of Fas2 inhibition by studying hydrolysis of cationic and neutral substrates and by determining the kinetics of interaction for fast equilibrating cationic and neutral reversible inhibitors with the AChE.Fas2 complex and free AChE. Catalytic parameters, derived by eliminating residual Fas2-resistant activity, reveal that Fas2 reduces k(cat)/K(m) up to 10(6)-fold for cationic substrates and less than 10(3)-fold for neutral substrates. Rate constants for association of reversible inhibitors with the active center of the AChE.Fas2 complex were reduced about 10(4)-fold for both cationic and neutral inhibitors, while dissociation rate constants were reduced 10(2)-to 10(3)-fold, compared with AChE alone. Rates of ligand association with both AChE and AChE.Fas2 complex were dependent on the protonation state of ionizable ligands but were also markedly reduced by protonation of enzyme residue(s) with pK(a) of 6.1-6.2. Linear free energy relationships between the equilibrium constant and the kinetic constants show that Fas2, presumably through an allosteric influence, markedly alters the position of the transition state in the reaction pathway. Since Fas2 complexation introduces an energetic barrier for hydrolysis of substrates that exceeds that found for association of reversible ligands, Fas2 influences catalytic parameters by a more complex mechanism than simple restriction of diffusional entry and exit from the active center. Conformational flexibility appears critical for facilitating ligand passage in the narrow active center gorge for both AChE and the AChE.Fas2 complex. PMID- 11036077 TI - Outside-in signaling pathway linked to CD146 engagement in human endothelial cells. AB - CD146 (S-Endo 1 Ag or MUC18) is a transmembrane glycoprotein expressed on endothelial cells on the whole vascular tree. CD146 is located at the intercellular junction where it plays a role in the cohesion of the endothelial monolayer. CD146 engagement initiates an outside-in signaling pathway involving the protein tyrosine kinases FYN and FAK as well as paxillin. Here we report that CD146 engagement by its specific monoclonal antibody in human umbilical vein endothelial cells induces a Ca(2+) influx that is sensitive to thapsigargin and EGTA treatment, indicating that CD146 engagement initiates a store-operated calcium mobilization. In addition, biochemical and pharmacological analysis revealed that CD146 engagement initiates the tyrosine phosphorylation of phospholipase C-gamma, Pyk2, and p130(Cas). Pharmacological inhibition of Ca(2+) flux with 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acetoxymethyl ester and EGTA indicated that an increase in Ca(2+) is required for Pyk2 and p130(Cas) tyrosine phosphorylation. Moreover, a complex association was observed between Pyk2, p130(Cas), and paxillin. These results indicate that CD146 is coupled to a FYN-dependent pathway that triggers Ca(2+) flux via phospholipase C-gamma activation leading subsequently to the tyrosine phosphorylation of downstream targets such as Pyk2, p130(Cas), FAK, and paxillin. In addition to its role in cell-cell adhesion, CD146 is a signaling molecule involved in the dynamics of actin cytoskeleton rearrangement. PMID- 11036078 TI - Evidence for two distinct epitopes within collagen for activation of murine platelets. AB - It has recently been shown that the monoclonal antibody JAQ1 to murine glycoprotein VI (GPVI) can cause aggregation of mouse platelets upon antibody cross-linking and that collagen-induced platelet aggregation can be inhibited by preincubation of platelets with JAQ1 in the absence of cross-linking (Nieswandt, B., Bergmeier, W., Schulte, V., Rackebrandt, K., Gessner, J. E., and Zirngibl, H. (2000) J. Biol. Chem. 275, 23998-24002). In the present study, we have shown that cross-linking of GPVI by JAQ1 results in tyrosine phosphorylation of the same profile of proteins as that induced by collagen, including the Fc receptor (FcR) gamma-chain, Syk, LAT, SLP-76, and phospholipase C gamma 2. In contrast, platelet aggregation and tyrosine phosphorylation of these proteins were inhibited when mouse platelets were preincubated with JAQ1 in the absence of cross-linking and were subsequently stimulated with a collagen-related peptide (CRP) that is specific for GPVI and low concentrations of collagen. However, at higher concentrations of collagen, but not CRP, aggregation of platelets and tyrosine phosphorylation of the above proteins (except for the adapter LAT) is re established despite the presence of JAQ1. These observations suggest that a second activatory binding site, which is distinct from the CRP binding site on GPVI on mouse platelets, is occupied in the presence of high concentrations of collagen. Although this could be a second site on GPVI that is activated by a novel motif within the collagen molecule, the absence of LAT phosphorylation in response to collagen in the presence of JAQ1 suggests that this is more likely to be caused by activation of a second receptor that is also coupled to the FcR gamma-chain. The possibility that this response is mediated by a receptor that is not coupled to FcR gamma-chain is excluded on the grounds that aggregation is absent in platelets from FcR gamma-chain-deficient mice. PMID- 11036079 TI - Hsp90 regulates p50(cdc37) function during the biogenesis of the activeconformation of the heme-regulated eIF2 alpha kinase. AB - Recent studies indicate that p50(cdc37) facilitates Hsp90-mediated biogenesis of certain protein kinases. In this report, we examined whether p50(cdc37) is required for the biogenesis of the heme-regulated eIF2 alpha kinase (HRI) in reticulocyte lysate. p50(cdc37) interacted with nascent HRI co-translationally and this interaction persisted during the maturation and activation of HRI. p50(cdc37) stimulated HRI's activation in response to heme deficiency, but did not activate HRI per se. p50(cdc37) function was specific to immature and inactive forms of the kinase. Analysis of mutant Cdc37 gene products indicated that the N-terminal portion of p50(cdc37) interacted with immature HRI, but not with Hsp90, while the C-terminal portion of p50(cdc37) interacted with Hsp90. The Hsp90-specific inhibitor geldanamycin disrupted the ability of both Hsp90 and p50(cdc37) to bind HRI and promote its activation, but did not disrupt the native association of p50(cdc37) with Hsp90. A C-terminal truncated mutant of p50(cdc37) inhibited HRI's activation, prevented the interaction of Hsp90 with HRI, and bound to HRI irrespective of geldanamycin treatment. Additionally, native complexes of HRI with p50(cdc37) were detected in cultured K562 erythroleukemia cells. These results suggest that p50(cdc37) provides an activity essential to HRI biogenesis via a process regulated by nucleotide-mediated conformational switching of its partner Hsp90. PMID- 11036080 TI - A set of Hox proteins interact with the Maf oncoprotein to inhibit its DNA binding, transactivation, and transforming activities. AB - Maf oncoprotein is a basic-leucine zipper (bZip) type of transcriptional activator. Since many transcription factors are known to form functional complexes, we searched for proteins that interact with the DNA-binding domain of Maf using the phage display method and identified two homeodomain-containing proteins, Hoxd12 and MHox/Prx1/Phox1/Pmx1. Studies with mutants of Hox and Maf proteins showed that they associate through their DNA-binding domains; the homeodomain of Hox and the bZip domain of Maf, respectively. Reflecting the high similarity of the bZip domain, all other Maf family members tested (c-/v-Maf, MafB, MafK, MafF, and MafG) also associated with the Hox proteins. Pax6, whose homeodomain is relatively similar to MHox, also could interact with Maf. However, two other bZip oncoproteins, Fos and Jun, failed to associate with the Hox proteins, while a distantly related Hox family member, Meis1, could not interact with Maf. Through interactions with the bZip domain, the Hox proteins inhibited the DNA binding activity of Maf, whereas the binding of Hox proteins to their recognition sequences was not abrogated by Maf. We further showed that coexpression of the Hox proteins repressed transcriptional activation and transforming activity of Maf. These results suggested that the interaction of a set of Hox proteins with Maf family members may interfere not only with their oncogenicity but also with their physiological roles. PMID- 11036081 TI - DNA melting within a binary sigma(54)-promoter DNA complex. AB - The final sigma(54) subunit of the bacterial RNA polymerase requires the action of specialized enhancer-binding activators to initiate transcription. Here we show that final sigma(54) is able to melt promoter DNA when it is bound to a DNA structure representing the initial nucleation of DNA opening found in closed complexes. Melting occurs in response to activator in a nucleotide-hydrolyzing reaction and appears to spread downstream from the nucleation point toward the transcription start site. We show that final sigma(54) contains some weak determinants for DNA melting that are masked by the Region I sequences and some strong ones that require Region I. It seems that final sigma(54) binds to DNA in a self-inhibited state, and one function of the activator is therefore to promote a conformational change in final sigma(54) to reveal its DNA-melting activity. Results with the holoenzyme bound to early melted DNA suggest an ordered series of events in which changes in core to final sigma(54) interactions and final sigma(54)-DNA interactions occur in response to activator to allow final sigma(54) isomerization and the holoenzyme to progress from the closed complex to the open complex. PMID- 11036082 TI - Biochemical basis for depressed serum retinol levels in transthyretin-deficient mice. AB - Transthyretin (TTR) acts physiologically in the transport of retinol in the circulation. We previously reported the generation and partial characterization of TTR-deficient (TTR(-)) mice. TTR(-) mice have very low circulating levels of retinol and its specific transport protein, retinol-binding protein (RBP). We have examined the biochemical basis for the low plasma retinol-RBP levels. Cultured primary hepatocytes isolated from wild type (WT) and TTR(-) mice accumulated RBP in their media to an identical degree, suggesting that RBP was being secreted from the hepatocytes at the same rate. In vivo experiments support this conclusion. For the first 11 h after complete nephrectomy, the levels retinol and RBP rose in the circulations of WT and TTR(-) mice at nearly identical rates. However, human retinol-RBP injected intravenously was more rapidly cleared from the circulation (t(12) = 0.5 h for TTR(-) versus t(12) >6 h for WT) and accumulated faster in the kidneys of TTR(-) compared with WT mice. The rate of infiltration of the retinol-RBP complex from the circulation to tissue interstitial fluids was identical in both strains. Taken together, these data indicate that low circulating retinol-RBP levels in TTR(-) mice arise from increased renal filtration of the retinol-RBP complex. PMID- 11036083 TI - The yeast NuA4 and Drosophila MSL complexes contain homologous subunits important for transcription regulation. AB - In Drosophila, the MSL complex is required for the dosage compensation of X linked genes in males and contains a histone acetyltransferase, MOF. A point mutation in the MOF acetyl-CoA-binding site results in male-specific lethality. Yeast Esa1p, a MOF homolog, is essential for cell cycle progression and is the catalytic subunit of the NuA4 acetyltransferase complex. Here we report that NuA4 purified from yeast with a point mutation in the acetyl-CoA-binding domain of Esa1p exhibits a strong decrease in histone acetyltransferase activity, yet has no effect on growth. We demonstrate that Eaf3p (Esa1p-associated factor-3 protein), a yeast protein homologous to the Drosophila dosage compensation protein MSL3, is also a stable component of the NuA4 complex. Unlike other subunits of the complex, it is not essential, and the deletion mutant has no growth phenotype. NuA4 purified from the mutant strain has a decreased apparent molecular mass, but retains wild-type levels of histone H4 acetyltransferase activity. The EAF3 deletion and the ESA1 mutation lead to a decrease in PHO5 gene expression; the EAF3 deletion also significantly reduces HIS4 and TRP4 expressions. These results, together with those previously obtained with both the MSL and NuA4 complexes, underscore the importance of targeted histone H4 acetylation for the gene-specific activation of transcription. PMID- 11036084 TI - Identification of an inhibitor of hsc70-mediated protein translocation and ATP hydrolysis. AB - Members of the hsc70 family of molecular chaperones are critical players in the folding and quality control of cellular proteins. Because several human diseases arise from defects in protein folding, the activity of hsc70 chaperones is a potential therapeutic target for these disorders. By using a known hsc70 modulator, 15-deoxyspergualin, as a seed, we identified a novel inhibitor of hsc70 activity. This compound, R/1, inhibits the endogenous and DnaJ-stimulated ATPase activity of hsc70 by 48 and 51%, respectively, and blocks the hsc70 mediated translocation of a preprotein into yeast endoplasmic reticulum-derived microsomal vesicles. Biochemical studies demonstrate that R/1 most likely exerts these effects by altering the oligomeric state of hsc70. PMID- 11036085 TI - Prothymosin alpha functions as a cellular oncoprotein by inducing transformation of rodent fibroblasts in vitro. AB - Prothymosin alpha (ProTalpha), a cellular molecule known to be associated with cell proliferation, is transcriptionally up-regulated on expression of c-myc and interacts with histones in vitro and associates with histone H1 in cells. Previous studies have also shown that ProTalpha is involved in chromatin remodeling. Recent studies have shown that ProTalpha interacts with the acetyl transferase p300 and an essential Epstein-Barr virus protein, EBNA3C, involved in regulation of viral and cellular transcription. These studies suggest a potential involvement in regulation of histone acetylation through the association with these cellular and viral factors. In the current studies, we show that heterologous expression of ProTalpha in the Rat-1 rodent fibroblast cell line results in increased proliferation, loss of contact inhibition, anchorage independent growth, and decreased serum dependence. These phenotypic changes seen in transfected Rat-1 cells are similar to those observed with a known oncoprotein, Ras, expressed under the control of a heterologous promoter and are characteristic oncogenic growth properties. These results demonstrate that the ProTalpha gene may function as an oncogene when stably expressed in Rat-1 cells and may be an important downstream cellular target for inducers of cellular transformation, which may include Epstein-Barr virus and c-myc. PMID- 11036086 TI - Crystal structure of the C-type lectin-like domain from the human hematopoietic cell receptor CD69. AB - CD69, one of the earliest specific antigens acquired during lymphoid activation, acts as a signal-transducing receptor involved in cellular activation events, including proliferation and induction of specific genes. CD69 belongs to a family of receptors that modulate the immune response and whose genes are clustered in the natural killer (NK) gene complex. The extracellular portion of these receptors represent a subfamily of C-type lectin-like domains (CTLDs), which are divergent from true C-type lectins and are referred to as NK-cell domains (NKDs). We have determined the three-dimensional structure of human CD69 NKD in two different crystal forms. CD69 NKD adopts the canonical CTLD fold but lacks the features involved in Ca(2+) and carbohydrate binding by C-type lectins. CD69 NKD dimerizes noncovalently, both in solution and in crystalline state. The dimer interface consists of a hydrophobic, loosely packed core, surrounded by polar interactions, including an interdomain beta sheet. The intersubunit core shows certain structural plasticity that may facilitate conformational rearrangements for binding to ligands. The surface equivalent to the binding site of other members of the CTLD superfamily reveals a hydrophobic patch surrounded by conserved charged residues that probably constitutes the CD69 ligand-binding site. PMID- 11036087 TI - Heterogeneous distribution of isoactins in cultured vascular smooth muscle cells does not reflect segregation of contractile and cytoskeletal domains. AB - We have previously demonstrated that alpha-smooth muscle (alpha-SM) actin is predominantly distributed in the central region and beta-non-muscle (beta-NM) actin in the periphery of cultured rabbit aortic smooth muscle cells (SMCs). To determine whether this reflects a special form of segregation of contractile and cytoskeletal components in SMCs, this study systematically investigated the distribution relationship of structural proteins using high-resolution confocal laser scanning fluorescent microscopy. Not only isoactins but also smooth muscle myosin heavy chain, alpha-actinin, vinculin, and vimentin were heterogeneously distributed in the cultured SMCs. The predominant distribution of beta-NM actin in the cell periphery was associated with densely distributed vinculin plaques and disrupted or striated myosin and alpha-actinin aggregates, which may reflect a process of stress fiber assembly during cell spreading and focal adhesion formation. The high-level labeling of alpha-SM actin in the central portion of stress fibers was related to continuous myosin and punctate alpha-actinin distribution, which may represent the maturation of the fibrillar structures. The findings also suggest that the stress fibers, in which actin and myosin filaments organize into sarcomere-like units with alpha-actinin-rich dense bodies analogous to Z-lines, are the contractile structures of cultured SMCs that link to the network of vimentin-containing intermediate filaments through the dense bodies and dense plaques. PMID- 11036088 TI - Transduction of TAT-HA-beta-galactosidase fusion protein into salivary gland derived cells and organ cultures of the developing gland, and into rat submandibular gland in vivo. AB - We have studied the transduction of TAT-HA-beta-galactosidase fusion protein into two cell lines of rat salivary gland origin, A5 and C6-21, into cells of fetal mouse submandibular glands in organ culture, and into rat submandibular gland after retrograde duct injection, using a histochemical method to demonstrate beta galactosidase activity. Transduction of the fusion protein into A5 and C6-21 cells was concentration- and time-dependent. Therefore, the intensity of the beta galactosidase staining, which was cytoplasmic, was less after 1 hr of exposure compared to exposures up to 24 hr. However, the fusion protein was transduced into 100% of both types of cultured cells. When explants of mouse fetuses at 13 days of gestation were exposed to the fusion proteins, both epithelial and mesenchymal cells were stained for the enzyme, with a conspicuous accumulation of the reaction product at perinuclear cytoplasmic regions. The histochemical staining of the mesenchymal cells was more intense compared to that seen in epithelial cells. TAT-HA-beta-galactosidase fusion protein was also delivered to rat submandibular glands by retrograde duct injection. Histochemical staining for beta-galactosidase activity of cryostat sections prepared from the injected glands revealed that the transduction of the fusion protein was also time- and dose-dependent. In the glands of rats sacrificed from 10 min to 1 hr after the retrograde injection, essentially all acinar and duct cells showed cytoplasmic staining. The intensity of the staining then declined, and was not seen in the glands of rats killed 24 hr after the injection of the fusion proteins. These results indicate that a full-length, active TAT fusion protein can be targeted to salivary gland cells both in vitro and in vivo to analyze physiological, developmental, and pathophysiological processes. PMID- 11036089 TI - Regional expression and histological localization of cysteine sulfinate decarboxylase mRNA in the rat kidney. AB - Cysteine sulfinate decarboxylase (CSD) is the rate-limiting biosynthetic enzyme of the pathway that forms taurine, a putative osmolyte in the kidney, which was previously localized in various segments of the nephron. Although CSD is known to be expressed in whole kidney extracts, no information on CSD mRNA regional expression and histological localization is yet available. Western blotting and Northern blotting were performed in four dissected regions of the kidney using an antiserum against recombinant CSD and a [(32)P]-dCTP-labeled CSD cDNA probe, respectively. In situ hybridization was carried out using a [(35)S]-CTP-labeled CSD RNA probe. A single protein (53 kD) and a single mRNA (2.5 kb) were detected, both of which appeared to be most enriched in the outer stripe of the outer medulla. In situ hybridization of CSD mRNA showed strong labeling of the thick tubules in the outer stripe of the outer medulla and in cortical medullary rays that corresponded to the proximal straight tubules. The significance of this restricted expression of CSD is discussed in relationship to the data previously reported on the location of taurine and the location of the taurine transporter along the nephron. PMID- 11036090 TI - Distribution of group II phospholipase A2 protein and mRNA in rat tissues. AB - Group II phospholipase A2 (PLA2) is an acute-phase protein and an important component of the host defense against bacteria. In this study we investigated the distribution of PLA2 protein by immunohistochemistry and the distribution of mRNA of PLA2 by Northern blotting and in situ hybridization in rat tissues. PLA2 protein was localized in the Paneth cells of the intestinal mucosa, chondrocytes and the matrix of cartilage, and megakaryocytes in the spleen. By Northern blotting, mRNA of PLA2 was found in the gastrointestinal tract, lung, heart, and spleen. By in situ hybridization, PLA2 mRNA was localized in the Paneth cells of the small intestinal mucosa but in no other cell types. Our results show specific distribution of PLA2 in a limited number of cell types in rat tissues. The reagents developed in this study (the anti-rat PLA2 antibody and probes for Northern blotting and in situ hybridization of mRNA of rat PLA2) will provide useful tools for future studies concerning the role of PLA2 in various experimental disease models. PMID- 11036091 TI - Quantitation of plasma membrane expression of a fusion protein of Na/H exchanger NHE3 and green fluorescence protein (GFP) in living PS120 fibroblasts. AB - We developed a confocal morphometric analysis to quantitate the relative plasma membrane (PM) expression of the Na/H exchanger NHE3 in living PS120 fibroblasts. NHE3 is a membrane transport protein that is acutely regulated by changes in the number of molecules expressed at the PM. To quantitate the PM expression of NHE3 under various experimental conditions, we stably expressed a chimera of rabbit NHE3 and green fluorescent protein (NHE3-GFP) in PS120 fibroblasts. A three dimensional (3D) map of the intracellular distribution of NHE3-GFP was obtained by confocal laser scanning microscopy (CLSM) of cells superfused with a styryl dye, FM 4-64. This fluorophore rapidly and reversibly labeled the outer lipid layer of the PM, which allowed generation of a digital mask of the PM and calculation of the fraction of a total cellular NHE3-GFP expressed at the PM. This analysis was successfully used to quantitate the relative PM expression of NHE3-GFP in control cells (25%) and a decrease in the expression caused by subsequent exposure of cells to wortmannin (5.1%). Reliability of the method was confirmed by cell surface biotinylation, which yielded very similar results. Confocal morphometric analysis is fast and reproducible and could potentially be used for investigations on regulation of expression of other membrane proteins. PMID- 11036092 TI - Bone morphogenetic protein (BMP) localization in developing human and rat growth plate, metaphysis, epiphysis, and articular cartilage. AB - We assessed the distribution and relative staining intensity of bone morphogenetic protein (BMP)-1-7 by immunohistochemistry in tibial growth plates, epiphyses, metaphyses, and articular cartilage in one 21-week and one 22-week human fetus and in five 10-week-old Sprague-Dawley rats. In the rats, articular cartilage was also examined. BMP proteins were mostly cytoplasmic, with negligible matrix staining. Highest BMP levels were seen in (a) hypertrophic and calcifying zone chondrocytes of growth plate (BMP-1-7), (b) osteoblasts and/or osteoprogenitor fibroblasts and vascular cells of the metaphyseal cortex and medulla (BMP-1-6), (c) osteoclasts of the metaphysis and epiphysis (BMP-1,-4,-5, and -6), and (d) mid to deep zone articular chondrocytes of weanling rats (BMP-1 7). BMP staining in osteoclasts, an unexpected finding, was consistently strong with BMP-4, -5, and -6 but was variable and dependent on osteoclast location with BMP-2,-3, and -7. BMP-1-7 were moderately to intensely stained in vascular canals of human fetal epiphyseal cartilage by endothelial cells and pericytes. BMP-1,-3, 5,-6, and -7 were localized in hypertrophic chondrocytes adjacent to cartilage canals. We conclude that BMP expression is associated with maturing chondrocytes of growth plate and articular cartilage, and may play a role in chondrocyte differentiation and/or apoptosis. BMP appears to be expressed by osteoclasts and might be involved in the intercellular "cross-talk" between osteoclasts and neighboring osteoprogenitor cells at sites of bone remodeling. PMID- 11036093 TI - Bismuth autometallography: protocol, specificity, and differentiation. AB - We provide a detailed protocol of the autometallographic bismuth technique and evaluate the specificity of the technique. We show by the multi-element technique "proton-induced X-ray microanalysis" (PIXE) that the autometallographic grains contain silver, bismuth, and sulfur, proving that autometallography can be used for specific tracing of bismuth bound as bismuth sulfide clusters in tissue sections from Bi-exposed animals or humans. In sections from animals exposed concurrently to selenium and bismuth, the autometallographic grains also contain selenium. This demonstrates that, if present in excess in the organisms, selenium will bind to exogenous bismuth, creating bismuth selenide clusters. As a further possible control for specificity and as a tool for differentiating among autometallographically detectable metals in sections containing more than one, we describe how bismuth sulfide clusters can be removed from Epon-embedded tissue sections by potassium cyanide. PMID- 11036094 TI - Localization of transcription factor AP-1 family proteins in ameloblast nuclei of the rat incisor. AB - We examined by immunocytochemistry the localization of the AP-1 family proteins c Jun, JunB, JunD, c-Fos, FosB, Fra-1, and Fra-2 in rat incisor ameloblasts. Most of the antibodies against AP-1 family proteins, except for c-Fos-specific antibody, labeled ameloblast nuclei. The labeling intensity of the c-Jun, JunD, and Fra-2 antibodies was stronger than that of JunB, FosB, and Fra-1. Antibody reactivities of c-Jun, JunD, and Fra-2 were greatly enhanced during or after the transition zone. Furthermore, c-Jun antibodies labeled maturation ameloblasts in a cyclic pattern, which was correlated with ameloblast modulation. Disruption of ameloblast modulation by colchicine injection resulted in greatly decreased reactivity of the c-Jun antibody in the ameloblast nuclei of the maturation zone. Phospho-specific antibodies to c-Jun labeled ameloblast nuclei only weakly throughout the secretion, transition, and maturation zones. These results suggest that the stage-specific localization of AP-1 in ameloblasts is closely related to tooth enamel formation. PMID- 11036095 TI - Immunohistochemical detection of ribosomal transcription factor UBF and AgNOR staining identify apoptotic events in neoplastic cells of Hodgkin's disease and in other lymphoid cells. AB - Ribosomal RNA synthesis is a key molecular process for understanding the mechanisms that drive cell proliferation. In this process, the upstream binding factor (UBF) is involved in regulating rDNA transcription at the nucleolus, together with RNA polymerase I. Recently, UBF was demonstrated to be a substrate for selective cleavage by specific proteases during apoptosis. Here we studied the expression of UBF in several cases of Hodgkin's disease (HD) by immunostaining and found it to be absent or clearly diminished in a high proportion of Reed-Sternberg cells and Hodgkin cells compared to small reactive lymphocytes. This result contrasted with labeling of those cells by the AgNOR technique, a marker of cell proliferation dependent on increased amounts of several proteins related to ribosome assembly. Disappearance of UBF and preservation of other NOR proteins is consistent with the pattern of selective proteolysis by caspases described in early stages of apoptosis. This correlates well with our results observed on induction of apoptosis in Jurkat cells treated with anti-FAS/APO-1 serum and with those in aged germinal center B-cells, in which UBF was no longer seen although the staining signal of other NOR proteins was maintained. These results support the concept that the rate of apoptosis is higher in neoplastic cells of HD than in the benign reactive lymphocyte population. Differential proteolysis of NOR proteins, as revealed by double staining of UBF and AgNOR, may prove valuable for identification of early stages of apoptosis in cytological and histopathological samples. PMID- 11036096 TI - Myotube formation is delayed but not prevented in MyoD-deficient skeletal muscle: studies in regenerating whole muscle grafts of adult mice. AB - We compared the time course of myogenic events in vivo in regenerating whole muscle grafts in MyoD(-/-) and control BALB/c adult mice using immunohistochemistry and electron microscopy. Immunohistochemistry with antibodies to desmin and myosin revealed a striking delay by about 3 days in the formation of myotubes in MyoD(-/-) autografts compared with BALB/c mice. However, myotube formation was not prevented, and autografts in both strains appeared similar by 8 days. Electron microscopy confirmed myotube formation in 8- but not 5-day MyoD(-/-) grafts. This pattern was not influenced by cross-transplantation experiments between strains examined at 5 days. Antibodies to proliferating cell nuclear antigen demonstrated an elevated level of replication by MyoD(-/-) myoblasts in autografts, and replication was sustained for about 3 days compared with controls. These data indicate that the delay in the onset of differentiation and hence fusion is related to extended proliferation of the MyoD(-/-) myoblasts. Overall, although muscle regeneration was delayed it was not impaired in MyoD(-/ ) mice in this model. PMID- 11036097 TI - Tissue distribution of the lipocalin alpha-1 microglobulin in the developing human fetus. AB - Alpha-1 microglobulin (alpha(1)m), a lipocalin, is an evolutionarily conserved immunomodulatory plasma protein. In all species studied, alpha(1)m is synthesized by hepatocytes and catabolized in the renal proximal tubular cells. alpha(1)m deficiency has not been reported in any species, suggesting that its absence is lethal and indicating an important physiological role for this protein To clarify its functional role, tissue distribution studies are crucial. Such studies in humans have been restricted largely to adult fresh/frozen tissue. Formalin-fixed, paraffin-embedded multi-organ block tissue from aborted fetuses (gestational age range 7-22 weeks) was immunohistochemically examined for alpha(1)m reactivity. Moderate to strong reactivity was seen at all ages in hepatocytes, renal proximal tubule cells, and a subset of pancreatic islet cells. Muscle (cardiac, skeletal, or smooth), adrenal cortex, a scattered subset of intestinal mucosal cells, tips of small intestinal villi, and Leydig cells showed weaker and/or variable levels of reactivity. Connective tissue stained with variable location and intensity. The following cells/sites were consistently negative: thymus, spleen, hematopoietic cells, lung parenchyma, glomeruli, exocrine pancreas, epidermis, cartilage/bone, ovary, seminiferous tubules, epididymis, thyroid, and parathyroid. The results underscore the dominant role of liver and kidney in fetal alpha(1)m metabolism and provide a framework for understanding the functional role of this immunoregulatory protein. PMID- 11036098 TI - A highly sensitive quantitative cytosensor technique for the identification of receptor ligands in tissue extracts. AB - Because G-protein-coupled receptors (GPCRs) constitute excellent putative therapeutic targets, functional characterization of orphan GPCRs through identification of their endogenous ligands has great potential for drug discovery. We propose here a novel single cell-based assay for identification of these ligands. This assay involves (a) fluorescent tagging of the GPCR, (b) expression of the tagged receptor in a heterologous expression system, (c) incubation of the transfected cells with fractions purified from tissue extracts, and (d) imaging of ligand-induced receptor internalization by confocal microscopy coupled to digital image quantification. We tested this approach in CHO cells stably expressing the NT1 neurotensin receptor fused to EGFP (enhanced green fluorescent protein), in which neurotensin promoted internalization of the NT1 EGFP receptor in a dose-dependent fashion (EC(50) = 0.98 nM). Similarly, four of 120 consecutive reversed-phase HPLC fractions of frog brain extracts promoted internalization of the NT1-EGFP receptor. The same four fractions selectively contained neurotensin, an endogenous ligand of the NT1 receptor, as detected by radioimmunoassay and inositol phosphate production. The present internalization assay provides a highly specific quantitative cytosensor technique with sensitivity in the nanomolar range that should prove useful for the identification of putative natural and synthetic ligands for GPCRs. PMID- 11036099 TI - Transient expression of transglutaminase C during prenatal development of human muscles. AB - Tissue transglutaminase (TGase C, TGase II) is known to participate in cellular processes during morphogenesis, differentiation, and development of various prenatal tissues and organs. The expression of TGase C during myoblast proliferation and attachment to external laminae was examined by immunohistochemical (IH) localization at 5-12 weeks of developmental stages of prenatal human muscle in 23 embryos. IH detection using a monospecific antibody to TGase C showed a prominent expression of TGase C in muscle cells as stage- and spatial-specific patterns during an early embryonal period. The myoblasts of intervertebral, tongue, and limb muscles, attached to adjacent cartilaginous skeletons or fibrous fascia, showed a pronounced expression of TGase C at 5-6, 6 7, and 7-8 weeks after fertilization, respectively. The most intense activity of TGase C was observed in some cardiac myoblasts infiltrating into endocardial mesenchyme at 6-7 weeks after fertilization. Although weak staining was detected until 14 weeks after fertilization, the level of TGase C expression in all muscles was significantly decreased after 6-7 weeks, with the exception that the smooth muscle cells of blood vessels and gastrointestinal tract showed diffusely intense staining of TGase C between 5 and 12 weeks after fertilization. Western blotting analysis of the cellular extracts of pooled samples showed a single strong band at 80 kD at 6 weeks after fertilization. This band became weaker after 8-10 weeks of prenatal development. These findings of transient expression of TGase C, which coincides with the development of myoblast anchoring and differentiation, suggest that TGase C plays a role in myoblast attachment to the extracellular laminae during the early embryonal period. PMID- 11036100 TI - Methods for imaging labeled neurons together with neuropil features in Drosophila. AB - We describe staining protocols for serial semithin sections of Drosophila central ganglia that allow visualization of gene expression in particular neurons with counterstaining to display the ganglion architecture. Green fluorescent protein (GFP), expressed in a subset of sensory neurons from a selected enhancer trap line, is visualized by conventional immunohistochemistry with a peroxidase-linked antibody, and neural architecture is revealed by reduced silver staining. This makes visible in histological sections the same GFP-labeled cells seen with confocal microscopy, but with the especial advantage that neuropil structures are also revealed at the level of individual cells and neuron processes. Not only does this allow the physical relationships among intracellularly labeled neurons to be determined by reference to specific features in the neuropil but it also enables a function to be ascribed provisionally to particular regions of neuropil. These methods have particular utility for mapping morphological information on specific neurons in the context of central nervous system architecture, both in adult Drosophila and during development. PMID- 11036101 TI - Dragons 'round the fleece again: STI571 versus alpha1 acid glycoprotein. PMID- 11036102 TI - Adenosine triphosphate-binding cassette proteins and bioavailability: "we can pump you up (or out)". PMID- 11036103 TI - After 40 years, mammography remains as much emotion as science. PMID- 11036104 TI - Oncologist's role critical to clinical trial enrollment. PMID- 11036105 TI - Stat bite: Cervical cancer incidence and mortality in U.S. blacks and whites, 1973-1997. PMID- 11036106 TI - Efforts aimed at risk communication flourish. PMID- 11036107 TI - Sunscreen: bodies of research remain to be covered. PMID- 11036108 TI - Camps bring adventure to children with cancer. PMID- 11036109 TI - Role of alpha1 acid glycoprotein in the in vivo resistance of human BCR-ABL(+) leukemic cells to the abl inhibitor STI571. AB - BACKGROUND: Chronic myeloid leukemia is caused by a chromosomal translocation that results in an oncogenic fusion protein, Bcr-Abl. Bcr-Abl is a tyrosine kinase whose activity is inhibited by the antineoplastic drug STI571. This drug can cure mice given an injection of human leukemic cells, but treatment ultimately fails in animals that have large tumors when treatment is initiated. We created a mouse model to explore the mechanism of resistance in vivo. METHODS Nude mice were injected with KU812 Bcr-Abl(+) human leukemic cells. After 1 day (no evident tumors), 8 days, or 15 days (tumors >1 g), mice were treated with STI571 (160 mg/kg every 8 hours). Cells recovered from relapsing animals were used for in vitro experiments. Statistical tests were two-sided. RESULTS: Tumors regressed initially in all STI571-treated mice, but all mice treated 15 days after injection of tumor cells eventually relapsed. Relapsed animals did not respond to further STI571 treatment, and their Bcr-Abl kinase activity in vivo was not inhibited by STI571, despite high plasma concentrations of the drug. However, tumor cells from resistant animals were sensitive to STI571 in vitro, suggesting that a molecule in the plasma of relapsed animals may inactivate the drug. The plasma protein alpha1 acid glycoprotein (AGP) bound STI571 at physiologic concentrations in vitro and blocked the ability of STI571 to inhibit Bcr-Abl kinase activity in a dose-dependent manner. Plasma AGP concentrations were strongly associated with tumor load. Erythromycin competed with STI571 for AGP binding. When animals bearing large tumors were treated with STI571 alone or with a combination of STI571 and erythromycin, greater tumor reductions and better long-term tumor-free survival (10 of 12 versus one of 13 at day 180; P:<.001) were observed after the combination treatment. CONCLUSION: AGP in the plasma of relapsed animals binds to STI571, preventing this compound from inhibiting the Bcr/Abl tyrosine kinase. Molecules such as erythromycin that compete with STI571 for binding to AGP may enhance the therapeutic potential of this drug. PMID- 11036110 TI - Role of breast cancer resistance protein in the bioavailability and fetal penetration of topotecan. AB - BACKGROUND AND METHODS: Breast cancer resistance protein (BCRP/MXR/ABCP) is a multidrug-resistance protein that is a member of the adenosine triphosphate binding cassette family of drug transporters. BCRP can render tumor cells resistant to the anticancer drugs topotecan, mitoxantrone, doxorubicin, and daunorubicin. To investigate the physiologic role of BCRP, we used polarized mammalian cell lines to determine the direction of BCRP drug transport. We also used the BCRP inhibitor GF120918 to assess the role of BCRP in protecting mice against xenobiotic drugs. Bcrp1, the murine homologue of BCRP, was expressed in the polarized mammalian cell lines LLC-PK1 and MDCK-II, and the direction of Bcrp1-mediated transport of topotecan and mitoxantrone was determined. To avoid the confounding drug transport provided by P-glycoprotein (P-gp), the roles of Bcrp1 in the bioavailability of topotecan and the effect of GF120918 were studied in both wild-type and P-gp-deficient mice and their fetuses. RESULTS: Bcrp1 mediated apically directed transport of drugs in polarized cell lines. When both topotecan and GF120918 were administered orally, the bioavailability (i.e., the extent to which a drug becomes available to a target tissue after administration) of topotecan in plasma was dramatically increased in P-gp-deficient mice (greater than sixfold) and wild-type mice (greater than ninefold), compared with the control (i.e., vehicle-treated) mice. Furthermore, treatment with GF120918 decreased plasma clearance and hepatobiliary excretion of topotecan and increased (re-)uptake by the small intestine. In pregnant GF120918-treated, P-gp-deficient mice, relative fetal penetration of topotecan was twofold higher than that in pregnant vehicle-treated mice, suggesting a function for BCRP in the maternal fetal barrier of the placenta. CONCLUSIONS: Bcrp1 mediates apically directed drug transport, appears to reduce drug bioavailability, and protects fetuses against drugs. We propose that strategic application of BCRP inhibitors may thus lead to more effective oral chemotherapy with topotecan or other BCRP substrate drugs. PMID- 11036111 TI - Predicting the cumulative risk of false-positive mammograms. AB - BACKGROUND: The cumulative risk of a false-positive mammogram can be substantial. We studied which variables affect the chance of a false-positive mammogram and estimated cumulative risks over nine sequential mammograms. METHODS: We used medical records of 2227 randomly selected women who were 40-69 years of age on July 1, 1983, and had at least one screening mammogram. We used a Bayesian discrete hazard regression model developed for this study to test the effect of patient and radiologic variables on a first false-positive screening and to calculate cumulative risks of a false-positive mammogram. RESULTS: Of 9747 screening mammograms, 6. 5% were false-positive; 23.8% of women experienced at least one false-positive result. After nine mammograms, the risk of a false positive mammogram was 43.1% (95% confidence interval [CI] = 36.6%-53.6%). Risk ratios decreased with increasing age and increased with number of breast biopsies, family history of breast cancer, estrogen use, time between screenings, no comparison with previous mammograms, and the radiologist's tendency to call mammograms abnormal. For a woman with highest-risk variables, the estimated risk for a false-positive mammogram at the first and by the ninth mammogram was 98.1% (95% CI = 69.3%-100%) and 100% (95% CI = 99.9%-100%), respectively. A woman with lowest-risk variables had estimated risks of 0.7% (95% CI = 0.2%-1.9%) and 4.6% (95% CI = 1. 1%-12.5%), respectively. CONCLUSIONS: The cumulative risk of a false positive mammogram over time varies substantially, depending on a woman's own risk profile and on several factors related to radiologic screening. By the ninth mammogram, the risk can be as low as 5% for women with low-risk variables and as high as 100% for women with multiple high-risk factors. PMID- 11036112 TI - Passive smoking exposure and female breast cancer mortality. AB - BACKGROUND: Several studies have reported positive associations between environmental tobacco smoke (ETS) and increased risk of breast cancer. However, studies of active smoking and risk of breast cancer are equivocal and in general do not support a positive association. To try to resolve this paradox, we examined the association between breast cancer mortality and potential ETS exposure from spousal smoking in an American Cancer Society prospective study of U.S. adult women. METHODS: We assessed breast cancer death rates in a cohort of 146 488 never-smoking, single-marriage women who were cancer free at enrollment in 1982. Breast cancer death rates among women whose husbands smoked were compared with those among women married to men who had never smoked. Cox proportional hazards modeling was used to control for potential risk factors other than ETS exposure. RESULTS: After 12 years of follow-up, 669 cases of fatal breast cancer were observed in the cohort. Overall, we saw no association between exposure to ETS and death from breast cancer (rate ratio [RR] = 1.0; 95% confidence interval [CI] = 0.8-1.2). We did, however, find a small, not statistically significant increased risk of breast cancer mortality among women who were married before age 20 years to smokers (RR = 1. 2; 95% CI = 0.8-1.8). CONCLUSIONS: In contrast to the results of previous studies, this study found no association between exposure to ETS and female breast cancer mortality. The results of our study are particularly compelling because of its prospective design as compared with most earlier studies, the relatively large number of exposed women with breast cancer deaths, and the reporting of exposure by the spouse rather than by proxy. PMID- 11036113 TI - CYP17 promoter polymorphism and breast cancer in Australian women under age forty years. AB - BACKGROUND: The cytochrome P450c17alpha enzyme functions in the steroid biosynthesis pathway, and altered endogenous steroid hormone levels have been reported to be associated with a T to C polymorphism in the 5' promoter region of the CYP17 gene. Because steroid hormone exposure is known to influence breast cancer risk, we conducted a population-based, case-control-family study to assess the relationship between the CYP17 promoter polymorphism and early-onset breast cancer. METHODS: Case subjects under 40 years of age at diagnosis of a first primary breast cancer, population-sampled control subjects, and the relatives of both case and control subjects were interviewed to record family history of breast cancer and other risk factors. CYP17 genotype was determined in 369 case subjects, 284 control subjects, and 91 relatives of case subjects. Genotype distributions were compared by logistic regression, and cumulative risk was estimated by a modified segregation analysis. All statistical tests were two tailed. RESULTS: Compared with the TT genotype (i.e., individuals homozygous for the T allele), the TC genotype was not associated with increased breast cancer risk (P: =.7). Compared with the TT and TC genotypes combined, the CC genotype was associated with a relative risk of 1. 81 (95% confidence interval [CI] = 1.15 2.86; P: =.01) before adjustment for measured risk factors and 1.63 (95% CI = 1.00-2.64; P: =.05) after adjustment. There was an excess of CC genotypes in case subjects who had at least one affected first- or second-degree relative, compared with control subjects unstratified by family history of breast cancer (23% versus 11%; P: =.006), and these case subjects had a threefold to fourfold higher risk than women of other groups defined by genotype and family history of breast cancer. Analysis of breast cancer in first- and second-degree relatives of case subjects with the CC genotype, excluding two known carriers of a deleterious mutation in BRCA1 or BRCA2, gave a relative hazard in women with the CC genotype of 3.48 (95% CI = 1.13-10.74; P: =.04), which is equivalent to a cumulative risk of 16% to age 70 years. CONCLUSIONS: The CC genotype may modify the effect of other familial risk factors for early-onset breast cancer. PMID- 11036114 TI - Patient preferences for axillary dissection in the management of early-stage breast cancer. AB - BACKGROUND: Recent data on the value of adjuvant therapy in lymph node-negative breast cancer and promising early data on less invasive strategies for managing the axilla have raised questions about the appropriate role of axillary lymph node dissection (ALND) in the management of early-stage breast cancer. We sought to evaluate how women weigh potential benefits of ALND-prognostic information, enhanced local control, and tailored therapy-against the risks of long-term morbidity that are associated with the procedure. METHODS: We used hypothetical scenarios to survey 82 randomly selected women with invasive breast cancer who had been treated with ALND and 62 women at risk for invasive breast cancer by virtue of a history of ductal carcinoma in situ (DCIS) who had not undergone ALND. RESULTS: Women in both the invasive cancer and the DCIS groups required substantial improvements in local control of the cancer (5% and 15%, respectively) and overall survival (3% and 10%, respectively) before they would opt for this procedure. Women with invasive cancer would choose ALND if it had only a 1% chance of altering treatment recommendations, whereas DCIS subjects required a 25% chance. Sixty-eight percent and 29% of women in the invasive cancer and DCIS groups, respectively, would accepted a 40% risk of arm dysfunction to gain prognostic information that would not change treatment. CONCLUSIONS: For most subjects treated previously for invasive breast cancer and almost half those at risk of the disease, the potential benefits of ALND, particularly the value of prognostic information, were sufficient to outweigh the risks of morbidity. However, women varied considerably in their preferences, highlighting the need to tailor decisions regarding management of the axilla to individual patients' values. PMID- 11036115 TI - Mutational analysis of endothelial cells derived from von Hippel-Lindau-related renal cancer. PMID- 11036116 TI - Re: race and differences in breast cancer survival in a managed care population. PMID- 11036117 TI - Re: improving the cost-effectiveness of colorectal cancer screening. PMID- 11036118 TI - A comparison of glyburide and insulin in women with gestational diabetes mellitus. AB - BACKGROUND: Women with gestational diabetes mellitus are rarely treated with a sulfonylurea drug, because of concern about teratogenicity and neonatal hypoglycemia. There is little information about the efficacy of these drugs in this group of women. METHODS: We studied 404 women with singleton pregnancies and gestational diabetes that required treatment. The women were randomly assigned between 11 and 33 weeks of gestation to receive glyburide or insulin according to an intensified treatment protocol. The primary end point was achievement of the desired level of glycemic control. Secondary end points included maternal and neonatal complications. RESULTS: The mean (+/-SD) pretreatment blood glucose concentration as measured at home for one week was 114+/-19 mg per deciliter (6.4+/-1.1 mmol per liter) in the glyburide group and 116+/-22 mg per deciliter (6.5+/-1.2 mmol per liter) in the insulin group (P=0.33). The mean concentrations during treatment were 105+/-16 mg per deciliter (5.9+/-0.9 mmol per liter) in the glyburide group and 105+/-18 mg per deciliter (5.9+/-1.0 mmol per liter) in the insulin group (P=0.99). Eight women in the glyburide group (4 percent) required insulin therapy. There were no significant differences between the glyburide and insulin groups in the percentage of infants who were large for gestational age (12 percent and 13 percent, respectively); who had macrosomia, defined as a birth weight of 4000 g or more (7 percent and 4 percent); who had lung complications (8 percent and 6 percent); who had hypoglycemia (9 percent and 6 percent); who were admitted to a neonatal intensive care unit (6 percent and 7 percent); or who had fetal anomalies (2 percent and 2 percent). The cord-serum insulin concentrations were similar in the two groups, and glyburide was not detected in the cord serum of any infant in the glyburide group. CONCLUSIONS: In women with gestational diabetes, glyburide is a clinically effective alternative to insulin therapy. PMID- 11036119 TI - Markers of myocardial damage and inflammation in relation to long-term mortality in unstable coronary artery disease. FRISC Study Group. Fragmin during Instability in Coronary Artery Disease. AB - BACKGROUND: In patients with unstable coronary artery disease, there is a relation between the short-term risk of death and blood levels of troponin T (a marker of myocardial damage) and C-reactive protein and fibrinogen (markers of inflammation). Using information obtained during an extension of the follow-up period in the Fragmin during Instability in Coronary Artery Disease trial, we evaluated the usefulness of troponin T, C-reactive protein, and fibrinogen levels and other indicators of risk as predictors of the long-term risk of death from cardiac causes. METHODS: Levels of C-reactive protein and fibrinogen at enrollment and the maximal level of troponin T during the first 24 hours after enrollment were analyzed in 917 patients included in a clinical trial of low molecular-weight heparin in unstable coronary artery disease. The patients were followed for a mean of 37.0 months (range, 1.6 to 50.6). RESULTS: During follow up, 1.2 percent of the 173 patients with maximal blood troponin T levels of less than 0.06 microg per liter died of cardiac causes, as compared with 8.7 percent of the 367 patients with levels of 0.06 to 0.59 microg per liter and 15.4 percent of the 377 patients with levels of at least 0.60 microg per liter (P=0.007 and P=0.001, respectively). The rates of death from cardiac causes were 5.7 percent among the 314 patients with blood C-reactive protein levels of less than 2 mg per liter, 7.8 percent among the 294 with levels of 2 to 10 mg per liter, and 16.5 percent among the 309 with levels of more than 10 mg per liter (P=0.29 and P=0.001, respectively). The rates of death from cardiac causes were 5.4 percent among the 314 patients with blood fibrinogen levels of less than 3.4 g per liter, 12.0 percent among the 300 with levels of 3.4 to 3.9 g per liter, and 12.9 percent among the 303 with levels of at least 4.0 g per liter (P=0.004 and P=0.69, respectively). In a multivariate analysis, levels of troponin T and C reactive protein were independent predictors of the risk of death from cardiac causes. CONCLUSIONS: In unstable coronary artery disease, elevated levels of troponin T and C-reactive protein are strongly related to the long-term risk of death from cardiac causes. These markers are independent risk factors, and their effects are additive with respect to each other and other clinical indicators of risk. PMID- 11036120 TI - Lipoprotein-associated phospholipase A2 as an independent predictor of coronary heart disease. West of Scotland Coronary Prevention Study Group. AB - BACKGROUND: Chronic inflammation is believed to increase the risk of coronary events by making atherosclerotic plaques in coronary vessels prone to rupture. We examined blood constituents potentially affected by inflammation as predictors of risk in men with hypercholesterolemia who were enrolled in the West of Scotland Coronary Prevention Study, a trial that evaluated the value of pravastatin in the prevention of coronary events. METHODS: A total of 580 men who had had a coronary event (nonfatal myocardial infarction, death from coronary heart disease, or a revascularization procedure) were each matched for age and smoking status with 2 control subjects (total, 1160) from the same cohort who had not had a coronary event. Lipoprotein-associated phospholipase A2, C-reactive protein, and fibrinogen levels, and the white-cell count were measured at base line, along with other traditional risk factors. The association of these variables with the risk of coronary events was tested in regression models and by dividing the range of values according to quintiles. RESULTS: Levels of C-reactive protein, the white-cell count, and fibrinogen levels were strong predictors of the risk of coronary events; the risk in the highest quintile of the study cohort for each variable was approximately twice that in the lowest quintile. However, the association of these variables with risk was markedly attenuated when age, systolic blood pressure, and lipoprotein levels were included in multivariate models. Levels of lipoprotein-associated phospholipase A2 (platelet-activating factor acetylhydrolase), the expression of which is regulated by mediators of inflammation, had a strong, positive association with risk that was not confounded by other factors. It was associated with almost a doubling of the risk in the highest quintile as compared with the lowest quintile. CONCLUSIONS: Inflammatory markers are predictors of the risk of coronary events, but their predictive ability is attenuated by associations with other coronary risk factors. Elevated levels of lipoprotein-associated phospholipase A2 appear to be a strong risk factor for coronary heart disease, a finding that has implications for atherogenesis and the assessment of risk. PMID- 11036121 TI - Efficacy of mycophenolate mofetil in patients with diffuse proliferative lupus nephritis. Hong Kong-Guangzhou Nephrology Study Group. AB - BACKGROUND: The combination of cyclophosphamide and prednisolone is effective for the treatment of severe lupus nephritis but has serious adverse effects. Whether mycophenolate mofetil can be substituted for cyclophosphamide is not known. METHODS: In 42 patients with diffuse proliferative lupus nephritis we compared the efficacy and side effects of a regimen of prednisolone and mycophenolate mofetil given for 12 months with those of a regimen of prednisolone and cyclophosphamide given for 6 months, followed by prednisolone and azathioprine for 6 months. Complete remission was defined as a value for urinary protein excretion that was less than 0.3 g per 24 hours, with normal urinary sediment, a normal serum albumin concentration, and values for serum creatinine and creatinine clearance that were no more than 15 percent above the base-line values. Partial remission was defined as a value for urinary protein excretion that was between 0.3 and 2.9 g per 24 hours, with a serum albumin concentration of at least 30 g per liter. RESULTS: Eighty-one percent of the 21 patients treated with mycophenolate mofetil and prednisolone (group 1) had a complete remission, and 14 percent had a partial remission, as compared with 76 percent and 14 percent, respectively, of the 21 patients treated with cyclophosphamide and prednisolone followed by azathioprine and prednisolone (group 2). The improvements in the degree of proteinuria and the serum albumin and creatinine concentrations were similar in the two groups. One patient in each group discontinued treatment because of side effects. Infections were noted in 19 percent of the patients in group 1 and in 33 percent of those in group 2 (P = 0.29). Other adverse effects occurred only in group 2; they included amenorrhea (in 23 percent of the patients), hair loss (19 percent), leukopenia (10 percent), and death (10 percent). The rates of relapse were 15 percent and 11 percent, respectively. CONCLUSIONS: For the treatment of diffuse proliferative lupus nephritis, the combination of mycophenolate mofetil and prednisolone is as effective as a regimen of cyclophosphamide and prednisolone followed by azathioprine and prednisolone but is less toxic. PMID- 11036122 TI - Transmission of Histoplasma capsulatum by organ transplantation. PMID- 11036123 TI - Images in clinical medicine. Mullerian adenosarcoma and early pregnancy. PMID- 11036124 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 32-2000. A boy with vertebral compression fractures. PMID- 11036125 TI - Oral hypoglycemic drugs for gestational diabetes. PMID- 11036126 TI - Inflammatory markers of coronary risk. PMID- 11036127 TI - Treatment of lupus nephritis--a work in progress. PMID- 11036129 TI - Correction: Use of Colonoscopy to Screen Asymptomatic Adults for Colorectal Cancer. PMID- 11036128 TI - The Republican and Democratic candidates speak on health care. PMID- 11036130 TI - Is (18)F-fluorodeoxyglucose positron emission tomography in recurrent colorectal cancer a contribution to surgical decision making? AB - BACKGROUND: Accuracy of (18)F-fluorodeoxyglucose positron emission tomography (FDG-PET) and contribution to surgical decision making in recurrent or metastatic colorectal cancer were evaluated. METHODS: One hundred whole-body PET tests in colorectal cancer patients (1994 to 1998) were compared with computed tomography (CT), liver ultrasonography, and carcinoembryonic antigen (CEA) test. Mean follow up was 12 months. RESULTS: Sensitivity, specificity, and accuracy of FDG-PET for malignant findings were, respectively, 98%, 90% and 95%; for 87 CT scans, 91%, 72%, and 82%; for 98 CEA tests, 76%, 90%, and 82%; for detection of liver metastases with PET, 100%, 99%, and 99%; and for 68 ultrasound tests, 87%, 96%, and 93%. PET accuracy for local recurrence was 96%. Additional information was provided by PET in 86% of cases (abdomen, thorax, liver). PET influenced surgical decisions in 61% of cases. CONCLUSION: FDG-PET adds relevant accuracy to the conventional staging of patients with colorectal cancer and may cost-effectively help to select the appropriate treatment. PMID- 11036131 TI - Magnetic resonance angiography of aorto-iliac disease. AB - BACKGROUND: Four different techniques for aorto-iliac magnetic resonance angiography (MRA) were assessed for accuracy using a digital subtraction angiography (DSA) gold standard. Surgeons' confidence in their ability to generate treatment plans with MRA and DSA was assessed, in consultation with a radiologist. METHODS: Two different two-dimensional (2D) time-of-flight (TOF) sequences, a phase-contrast sequence, and a contrast-enhanced (CE) MRA sequence were used. Receiver operating characteristic (ROC) curves were plotted and areas (A(z)) calculated from radiologists' readings. Surgeons' confidence in their ability to utilize the images for treatment planning was assessed with a 5-point Likert scale. Thirty-six patients were evaluated. RESULTS: CE MRA had a sensitivity, specificity, and A(z) of.92,.93, and.96, respectively, for stenoses 50% or greater. CE MRA performed better than other sequences, but the improvement compared with gated 2D TOF was not statistically significant. Interobserver agreement for CE MRA and DSA yielded identical Kappa values. Surgeons were most confident in DSA, followed by CE MRA, which was significantly preferred to other techniques. CONCLUSIONS: CE MRA closely approximates DSA in terms of diagnostic accuracy. Surgeons considering treatment plans are confident in the CE MRA technique, relative to other MRA methods. PMID- 11036132 TI - Factors influencing survival after resection for periampullary neoplasms. AB - BACKGROUND: The purpose of this study was to determine predictors of survival after resection for periampullary neoplasms. METHODS: Over a 15-year period, 208 patients underwent laparotomy for periampullary neoplasms. Data were analyzed to assess predictors of survival. RESULTS: Pathologic examination showed pancreatic cancer (n = 136; 65%), ampullary cancer (n = 28; 13%), distal common bile duct cancer (n = 10; 5%), duodenal cancer (n = 4; 2%), neuroendocrine tumor (n = 11; 5%), cystadenocarcinoma (n = 4; 2%), cystadenoma (n = 5; 2%), and other (n = 10; 5%). A total of 129 patients underwent pancreatic resection (71 Whipples, 35 total pancreatectomies, 21 distal pancreatectomies, and 2 partial pancreatectomies) whereas 79 patients were found to be unresectable and underwent palliative bypass and/or biopsy. Median survival was 20.4 months for resectable patients versus 4.5 months for unresectable patients (P<0.001). Of the 129 resected patients, factors significantly (P<0.05) favoring long-term survival on univariate analysis included well-differentiated histology, common bile duct or ampullary adenocarcinoma, early stage, tumor diameter <2 cm, negative margins, and absence of lymph node metastases, perineural, or vascular invasion. Age, sex, race, and type of procedure had no influence on survival. On multivariate analysis, only tumor differentiation appeared independently related to survival. Using Kendall's tau analysis, tumor type and grade correlated significantly with all other predictors. CONCLUSIONS: Of all variables studied, tumor type and poor tumor differentiation in periampullary neoplasms appear to be markers that predict a constellation of other adverse findings. PMID- 11036133 TI - Enteral nutrition prolongs delayed gastric emptying in patients after Whipple resection. AB - BACKGROUND: Delayed gastric emptying is one of the most frequent postoperative complications after Whipple resection. In the present study we evaluated the role of enteral nutrition in the development of delayed gastric emptying after Whipple resection. PATIENTS AND METHODS: Between January 1996 and June 1998, 64 patients (30 female, 34 male) underwent a classic (n = 27) or pylorus-preserving (n = 37) Whipple resection. Two patients were excluded; 30 patients received enteral and 32 patients received no-enteral nutrition. RESULTS: Delayed gastric emptying occurred significantly more in patients with enteral (17 of 30, 57%) than in patients with no-enteral nutrition (5 of 32, 16%) (P <0.01). Consequently, patients in the enteral nutrition group had a nasogastric tube for a significantly (P<0.01) longer period and had a significantly (P<0.01) longer hospital stay than patients in the no-enteral nutrition group. There were no differences in the frequency of occurrence of other postoperative complications between patients with enteral and no-enteral nutrition. CONCLUSION: In patients undergoing a Whipple resection, enteral nutrition is associated with a higher frequency of delayed gastric emptying with no advantages regarding other postoperative complications and should therefore be restricted to specific indications. PMID- 11036134 TI - Simple versus double jejunal pouch for reconstruction after total gastrectomy. AB - BACKGROUND: Even though many types of reconstruction after total gastrectomy have been proposed to reduce postgastrectomy syndromes, choosing a method that would further improve the quality of life and nutrition of the gastrectomized patient is controversial. Hunt-Lawrence single pouch reconstruction seems to obtain better results compared with the more common Roux-en-Y technique, but both of these reconstructive approaches are associated with some reduction in food intake and some problems in achievement of ideal body weight. METHODS: In this prospective, randomized trial, after total gastrectomy 18 patients had reconstruction according to the Hunt-Lawrence or single pouch technique (SP group), whereas for 23 patients, the technique was modified with construction of a second pouch in the distal portion of the jejunal loop (DP group). Patients in the two groups were compared at 12 months after surgery for problems in gastrointestinal function, quality of life, improvement in body weight and nutritional parameters, serum albumin, hemoglobin level, and serum protein. RESULTS: The DP group demonstrated fewer symptom problems, better weight maintenance, and better laboratory values when compared with patients undergoing standard single jejunal pouch reconstruction. CONCLUSIONS: Reconstruction with use of a double pouch as a gastric substitute leads to better outcome assessments than with a single pouch reconstruction. Our double pouch technique has demonstrated significant improvement in quality of life and nutritional recovery in terms of functional results as well as patient satisfaction. PMID- 11036135 TI - Preemptive bupivacaine offers no advantages to postoperative wound infiltration in analgesia for outpatient breast biopsy. AB - BACKGROUND: Adequate analgesia is important after surgery and in particular after ambulatory surgery. Preemptive administration of analgesics, ie, prior to commencing surgery, has many theoretical advantages. METHODS: In this prospective randomized study, the use of preincisional bupivacaine was compared with a postincision dose for the relief of postoperative pain, in 74 patients undergoing day-case breast biopsy. RESULTS: Demographic criteria were similar in both groups. There were no differences in pain scores postoperatively on the visual analog scale (VAS): VAS at 30 minutes 4.5 ([SD] 2.4) versus 4.7 (1.9); P = not significant (NS); VAS at 60 minutes 3.3 (2. 3) versus 3.6 (2.2); P = NS; VAS at 120 minutes 1.9 (1.7) versus 2.5 (2.0); P = NS; VAS at 240 minutes 0.9 (1.0) versus 1.3 (1.4); P = NS. There was no difference in the number of patients requiring additional analgesia: 13 (36%) versus 18 (47%); P = NS. Nor was there a difference in the time to additional analgesia: 55.0 (37.8) versus 55.3 (39.2) minutes; P = NS. CONCLUSIONS: The administration of local anaesthesia prior to starting surgery does not appear to have any advantage over its postoperative administration in patients undergoing ambulatory breast biopsy. PMID- 11036136 TI - Etiology of small bowel obstruction. AB - BACKGROUND: Small bowel obstruction (SBO) is a major cause of morbidity and financial expenditure in hospitals around the world. The leading cause of SBO in the western world has become adhesions. The goal of this study was to determine the causes of SBO. METHODS: The medical records of all patients admitted to one hospital between 1986 and 1996 with the diagnosis of SBO were reviewed retrospectively. This included 552 patients accounting for 1,001 admissions. RESULTS: The etiology of SBO was adhesions (74%), Crohn's disease (7%), neoplasia (5%), hernia (2%), radiation (1%), and miscellaneous (11%). Patients with Crohn's disease were younger than patients with other etiologies. Surprisingly, recurrence rates were similar for patients treated operatively as for those treated nonoperatively with the exception in the hernia group where higher recurrence rates were noted for patients initially treated in a nonoperative manner. CONCLUSION: The most common cause of SBO is adhesions followed by Crohn's disease and neoplasia. PMID- 11036137 TI - Improvement of the Roux limb function using a new type of "uncut Roux" limb. AB - BACKGROUND: The Roux stasis syndrome is characterized by symptoms of upper gut stasis following Roux-en-Y gastrojejunostomy. The aim of this study was to compare a new type of uncut Roux-en-Y gastrojejunostomy with the conventional Roux-en-Y gastrojejunostomy after subtotal gastrectomy. METHODS: 51 patients (31 men and 20 women) had the conventional Roux-en-Y reconstruction and 54 patients (38 men and 16 women) had the new type of uncut Roux-en-Y reconstruction. The new type of uncut Roux-en-Y gastrojejunostomy consisted of an artificial jejunal occlusion and a short Roux limb (20 to 30 cm). RESULTS: The criteria included one of the four following conditions at the time of follow-up: chronic abdominal pain, postprandial fullness, persistent nausea, and intermittent vomiting that are worsened by eating. According to the criteria, the Roux stasis syndrome occurred in 19 patients (37.3%) with conventional Roux-en-Y reconstruction, and in 10 patients (18.5%) with uncut Roux-en-Y reconstruction (P = 0.033). CONCLUSIONS: A new type of Roux operation is able to alleviate not only the Roux stasis syndrome but also alkaline reflux gastritis or esophagitis by preserving motility of the Roux limb and diversion of duodenal juice from the gastric remnant. PMID- 11036138 TI - Quality of life before and after laparoscopic fundoplication. AB - BACKGROUND: Laparoscopic fundoplication is a well-established surgical option for the treatment of gastroesophageal reflux disease. The aim of this study was to assess the surgical outcomes from the patient's point of view by using a validated quality of life instrument. METHODS: Fifty patients have been prospectively included. All patients underwent a standardized 270-degree posterior fundoplication. Quality of life was measured by the Gastrointestinal Quality of Life Index (GIQLI), a 36-item-questionnaire. The patients received the questionnaire before surgery, and 3 months and 1 year after surgery. RESULTS: Preoperative score was 95.6+/-21 points. The score increased significantly (P <0.0005) at 3 months (103.6+/-16) and 1 year (111.4+/-22) after surgery. This improvement concerned the four domains of the questionnaire (symptoms, social functioning, physical status, and emotions). The score in patients at 1 year remained, however, significantly lower than that in healthy persons (126+/-18). CONCLUSIONS: GIQLI is a sensitive tool to assess surgical outcomes after fundoplication. The quality of life after surgery did not reach the level of healthy population, not because of failure of surgery to treat GERD but probably because of functional dyspepsia that was present prior to surgery and did not change after fundoplication. PMID- 11036139 TI - Chairpersons' opinions regarding quality control of surgical faculty performance in Japanese academic surgery departments. AB - BACKGROUND: The governance and power structure of the department of surgery depends to a large extent on the chairperson's decisions in Japanese medical schools. This paper reports the current collective opinions of surgery department chairpersons regarding the quality assessment of surgical faculty performance. METHODS: Surveyed were 78 chairpersons of general surgery departments from 72 Japanese medical schools. Chairpersons were questioned about administrative and organizational decision making: rank order requirements for full-time surgical faculties, coordination of staff for surgical operations, and performance outcome measures. RESULTS: In all, 68 (87%) chairpersons responded. When selecting surgical faculties, publishing competence (45%) and collaborative personality (44%) were the two foremost concerns of chairpersons. Teaching experience (0%) and board certification (2%) showed the lowest rate for the first priority among the 6 elements listed. The operator was mainly decided by the chairperson (63%) whereas the rest of the operative team members were decided by either the chairperson (28%), a specialty team (38%), or attending surgeons (32%). Thirty three chairpersons (49%) of 68 respondents used the morbidity and mortality conference as the only available approach for assessing surgical performance on a regular basis, whereas the remaining half did not have routine outcome measures. CONCLUSIONS: The results of this study indicate that surgery department chairpersons deemed collaborative personality and publishing competence the two major requirements for candidates of surgical faculties. Although the morbidity and mortality conference is currently the only available approach for assessing surgical performance, the majority of chairpersons felt that outcome measures should be based on more objective and structured criteria. PMID- 11036140 TI - Ileostomy construction in complex reoperative surgery with associated abdominal wall defects. AB - Patients who have undergone recent abdominal surgery and have generalized obliterative adhesions provide a major management problem if they require urgent ileostomy for sepsis or fistulization, particularly when there is an associated abdominal wall defect. This report describes a technique for creating an ileostomy through the skin and subcutaneous tissue only, medial to the fascial edge. This minimizes the intraoperative difficulties of stoma creation and reduces the dangers associated with mobilizing enough bowel to bring out through the fascia. PMID- 11036141 TI - Cystic adventitial disease of the popliteal artery. PMID- 11036142 TI - A simpler, less expensive technique for delayed primary closure of fasciotomies. AB - A variety of techniques have recently been advanced for delayed primary closure of wounds following emergent fasciotomy for compartment syndrome. We introduce a very simple, effective method for gradual reapproximation of margins using daily reapplication of Steri-strips (3M Surgical Products, St. Paul, Minnesota). This method allows final closure of fasciotomy wounds with simple suture in 5-8 days without scar contractures, marginal necrosis, infection, or significant pain. Moreover, because it requires no specialized equipment and can be applied in skilled nursing centers or at home by trained nurses, this technique could reduce the cost of caring for fasciotomy patients. PMID- 11036143 TI - A ten-year analysis of surgical education research. AB - BACKGROUND: Surgical education peer-reviewed publications have markedly increased over the last decade. The purpose of this study was to review the surgical education literature published over the last 10 years and address the following questions: What subjects in surgical education tend to be studied? What are the most to least commonly employed research designs and statistics? Has there been a change in how research data are collected? Where are these studies published? METHODS: A literature search encompassing surgical education papers published between January 1988 and August 1998 was performed. Four investigators coded qualifying abstracts on journal type, subject of research, data collection methods, research design, and statistics. Each investigator was asked to code 10 articles at the start of the study to assess interrater reliability. RESULTS: A total of 420 abstracts were evaluated. Interrater reliability yielded percent agreements ranging from 82% to 96%. Curriculum and teaching were the most frequent topics studied (40%), followed by assessment (23%) and program evaluation (18%). Most research designs used were descriptive (41%). Experimental design has progressively increased from 2% in 1988-89 to 16% in 1998. A total of 551 statistical methods were accounted for in the 420 abstracts. The most common statistical analyses used were descriptive statistics (32%). The predominant mode of data collection was through testing or direct observations (34%). Survey instruments followed closely as a popular data collection method at 27%. The majority of papers were published in peer-reviewed surgical journals (64%),followed by medical education journals (22%) and "other" journals (14%). CONCLUSIONS: An analysis of the surgical education literature demonstrates the growing emphasis on the use of educational research to explore relevant issues and problems. Descriptive research is most popular, with an increasing trend in experimental research. Publication of educational research in peer-reviewed surgical journals is becoming more popular. This study informs those interested in the surgical education research literature of current trends, and what they need to know for a more critical appraisal of this body of literature. PMID- 11036144 TI - Laparoscopic excision of accessory spleen. AB - BACKGROUND: Laparoscopic splenectomy has become an accepted procedure in the management of several hematologic diseases. Less clear is the effectiveness of laparoscopic excision of accessory spleens after initial splenectomy in the management of recurrent hematologic disease. We report here our early experience of this technique. METHODS: All patients who underwent laparoscopic excision of accessory spleens (LEAS) after initial splenectomy were reviewed for preoperative studies, technical success, and effects on either platelet count or hemoglobin level. RESULTS: In 5 patients LEAS was attempted. Two patients had initial open splenectomies, and 3 had initial laparoscopic splenectomies. Hematologic diagnoses were immune thrombocytopenic purpura (3), chronic lymphocytic leukemia induced thrombocytopenia (1), and autoimmune hemolytic anemia (1). All patients underwent preoperative damaged red blood cell scintigraphy, which demonstrated functioning splenic tissue, and abdominal computed tomography scans, which demonstrated a nodule in 4 of 5 patients. LEAS was technically successful in 4 patients, with the 1 failure also being the patient in whom the computed tomography scan could not demonstrate the accessory spleen. However, only 2 of the 4 patients after LEAS had durable hematologic responses to surgery, despite follow-up damaged red blood cell scintigraphy showing no residual functioning splenic tissue. CONCLUSION: LEAS can be technically successful when the accessory spleen is demonstrated on both damaged red blood cell scintigraphy and computed tomography scan; therefore, adequate visualization in both studies is required. However, hematologic response to excision may be less effective than with the initial splenectomy. Further study is needed to determine the causes of these outcomes. PMID- 11036145 TI - Inflammatory consequences of the translocation of bacteria and endotoxin to mesenteric lymph nodes. AB - BACKGROUND: Translocation of intestinal bacteria to mesenteric lymph nodes (MLNs) has been documented in humans under a variety of circumstances, yet its clinical significance remains to be established. The aim of this study was to correlate detectable translocation to MLNs of bacteria and endotoxin with local and systemic signs of inflammation. METHODS: From each of 10 patients with carcinoma of the cecal region two MLNs were harvested prior to resection. The presence of bacteria and endotoxin in the lymphatic tissue and blood was determined by culture methods and DNA preparation (PCR) and by a Limulus assay, respectively. Inflammatory mediators were determined in plasma and in MLN homogenates. RESULTS: Viable bacteria were detected in MLNs of 7 patients and in 9 of 20 lymph nodes. PCR revealed traces of bacteria in 4 patients and in 6 of their MLNs. Combining both modalities, the translocation rate was 80% and 55% for patients and MLNs, respectively. There was no detectable bacteremia. Endotoxin was found in the plasma of 7 patients and in 9 MLNs from 5 patients. There was no correlation between culture findings and endotoxin concentrations. Moreover, bacteriological data did not correspond to local or systemic inflammation. The group of MLN with detectable endotoxin differed significantly from LPS-negative nodes with respect to interleukin-6, interleukin-10, and sCD14. Systemic concentrations of endotoxin and inflammatory parameters did not correspond to levels within MLNs. CONCLUSION: Translocation to MLNs occurs in patients with cecal carcinoma. This, however, seems not to be of major clinical significance if no additional physiologic insults are encountered. Irrespective of the presence of bacteria, there are variations in inflammatory reactions between lymph nodes from one and the same patient, probably reflecting fluctuating response mechanisms to low-grade translocation. PMID- 11036146 TI - A nation's experience of bleeding complications during laparoscopy. AB - BACKGROUND: Bleeding complications during laparoscopic surgery are rare but probably underreported. The aim of the current study was to elucidate the clinical relevance of bleeding complications and major vascular injuries during standard laparoscopic procedures. PATIENTS AND METHODS: The Swiss Association of Laparoscopic and Thoracoscopic Surgery (SALTS) prospectively collected the data on 14,243 patients undergoing different standard laparoscopic procedures (1995 to 1997). These data were analyzed with special interest in intraoperative and postoperative bleeding complications and major vascular injuries. RESULTS: In all, 331 patients (2.3%) had intraoperative bleeding complications. Whereas 44 patients suffered from an external bleed of the abdominal wall, the bleeding was internal in the remaining 287. Thirty-three patients with internal bleeding required blood transfusion with a mean blood loss of 1,630 mL. Surgical hemostasis was necessary in 68% of external and 91% of internal bleeds. There were 250 patients (1.8%) with postoperative bleeding complications. External bleeding occurred in 143 patients, and 107 patients developed internal bleeding. External bleeding was mainly treated conservatively (92%), whereas 50% of internal bleeds required further surgical intervention. Major vascular injuries occurred in 12 patients (incidence 0.08%) with open treatment being necessary in all cases. CONCLUSIONS: Bleeding complications are, in fact, common during laparoscopic surgery. Meticulous dissection technique, immediate recognition, and adequate surgical treatment are mandatory for their management. PMID- 11036147 TI - The substrate for brain-stimulation reward in the lateral preoptic area. I. Anatomical mapping of its boundaries. AB - Given the putative role of the lateral preoptic area as a primary contributor of the cell bodies of origin of the descending pathway linking a subset of lateral hypothalamic and ventral tegmental area reward neurons, the distribution of self stimulation sites in this structure was mapped in 22 animals using moveable electrodes and threshold procedures. Ninety-seven electrode sites were evaluated with placements ranging from just rostral to the midline convergence of the anterior commissure back to the transition zone between the lateral preoptic and lateral hypothalamic areas; of these, roughly 2/3 supported self-stimulation which was widely observed throughout the lateral preoptic area and medial forebrain bundle. In general, self-stimulation thresholds obtained from lateral sites were most stable, and progressively so approaching more caudal regions. Examination of the slopes of the period/current trade-off functions revealed a tendency for higher values in lateral and caudal sites; in contrast, dorsoventral excursions did not influence these estimates. Taken together, these data provide support for the notion that the substrate for brain-stimulation reward in the lateral preoptic area has a relatively homogeneous distribution that is more diffusely organized than that found in reward sites activated further caudally in the medial forebrain bundle. PMID- 11036148 TI - The substrate for brain-stimulation reward in the lateral preoptic area. II. Connections to the ventral tegmental area. AB - This experiment investigated the existence of a direct anatomical connection between lateral preoptic and ventral tegmental areas that mediate brain stimulation reward using the behavioral adaptation of the collision test. This test is a double-pulse, two-electrode technique based on the axonal conduction failure that occurs when two separate sites in the same axon bundle are concurrently stimulated. This anatomical arrangement is inferred from the shape of the function relating the effectiveness of double-pulse stimulation to the interval between pulses. In this study, nine rats with a total of 44 pairs of sites were examined. In two pairs only was there a profile suggestive of an axonal collision effect, while the double-pulse effectiveness curve consistent with the properties of transynaptic collision was apparent for a single pair of sites; the remaining 93% were associated with relatively flat effectiveness curves. While electrode misalignment could be responsible for these results, there was adequate sampling to suggest that the preponderance of first stage signals that give rise to the rewarding effects mediated by the lateral preoptic and ventral tegmental areas do not travel along the same fiber bundle. However, stimulation applied to both sites concurrently produces a summation that is roughly 40% greater than stimulation at either site alone, suggesting reasonable integration of the reward signals generated by lateral preoptic and ventral tegmental area stimulation. PMID- 11036149 TI - In vivo study of tumor metabolism: an application of new multi-probe microdialysis system in the striatum of freely moving rats grafted with C6 cells. AB - The purpose of this study is to investigate the in vivo tumoral brain metabolism in free moving rats using microdialysis. Cells from C6 glioma cell line were inoculated in one striatum 15 days before the microdialysis experimentation. Then, using a new system allowing perfusion of several microdialysis probes in free moving rat, normalised dialysate levels of glucose, lactate and pyruvate were monitored in both glioma and control striatum. At the end of the procedure, animals were sacrificed for histological study. Data shows that probe functioning is similar in both tissues. The results for normalised glucose level were in striatum control: 2.14 mM, in tumoral striatum: 1.71 mM (P>0.1); for lactate, respectively, 0.86 and 1.65 mM (P<0.05) and for pyruvate, respectively, 65.56 and 140. 94 microM (P<0.05). This data clearly shows a significant increase of pyruvate and lactate in tumoral striatum compared to normal striatum, correlating previous in vitro studies on glioma metabolism. We conclude that this microdialysis technique is of value in tumoral brain and could constitute an interesting tool for a better understanding of glioma metabolism. PMID- 11036150 TI - Relations between the directions of vibration-induced kinesthetic illusions and the pattern of activation of antagonist muscles. AB - In humans, tendon vibration evokes illusory sensations of movement that are usually associated with an excitatory tonic response in muscles antagonistic to those vibrated (antagonist vibratory response, AVR), i.e., in the muscle groups normally contracted if the illusory movement had been performed. The aim of the present study was to investigate the relation between the parameters of the illusory sensation of movement and those of the AVR and to determine whether vectorial models could account for the integration of proprioceptive inputs from several muscles, as well as for the organization of the elementary motor commands leading to one unified motor response. For that purpose, we analyzed the relations between the anatomical site of the tendon vibration, the direction of the illusory movement, the muscles in which the AVR develops, and the characteristics of the AVR (surface EMG, motor unit types, firing rates, and activation latencies). This study confirmed the close relationship between the parameters of an AVR and those of the kinesthetic illusion. It showed that, during illusions of movements in different directions, motor units are activated according to a specific pattern correlated with their type, with the direction of the illusory movement and with the biomechanical properties of their bearing muscles. Finally, kinesthetic illusions and AVRs can be effectively represented using similar vectorial computations. These strong relations between the perceptual and motor effects of tendon vibration once again suggest that the AVR may result from a perceptual-to-motor transformation of proprioceptive information, rather than from spinal reflex mechanisms. PMID- 11036151 TI - Influence of corticotrophin releasing factor on neuronal cell death in vitro and in vivo. AB - Several studies have demonstrated that antagonists of the corticotrophin releasing factor (CRF) receptor markedly inhibit experimentally induced excitotoxic, ischaemic and traumatic brain injury in the rat, and that CRF expression is elevated in response to experimentally induced stroke or traumatic brain injury. CRF is also induced by the pro-inflammatory cytokine interleukin 1 (IL-1), which participates in various forms of neurodegeneration. The aim of this study was to test the hypothesis that CRF is toxic directly in vivo or in vitro. In primary cultures of rat cortical neurons, exposure to CRF (10 pM-100 nM) for 24 h failed to cause cell death directly, or to modify the neurotoxic effects of N-methyl-D-aspartate (NMDA). Similarly, infusion of CRF (0.3-5 microg) into specific brain regions of the rat did not induce cell death and did not significantly alter the neuronal damage produced by infusion of excitatory amino acids. These data demonstrate that CRF is not directly neurotoxic, and suggest that either CRF mediates neuronal damage by indirect actions (e.g. on the vasculature) and/or that CRF is not the endogenous ligand which contributes to neurodegeneration through activation of CRF receptors. PMID- 11036152 TI - Sodium-ascorbate cotransport controls intracellular ascorbate concentration in primary astrocyte cultures expressing the SVCT2 transporter. AB - Expression of the Na(+)-ascorbate cotransporter, SVCT2, was detected in rat brain and in primary cultures of cerebral astrocytes by Northern blot analysis. SVCT2 expression in cultured astrocytes increased in response to the cyclic AMP analog, dibutyryl cyclic AMP. A mathematical model of ascorbic acid transport was developed to evaluate the hypothesis that Na(+)-ascorbate cotransport across the plasma membrane regulates the steady state intracellular concentration of ascorbic acid in these cells. The outcomes predicted by this model were compared to experimental observations obtained with primary cultures of rat cerebral astrocytes exposed to normal and pathologic conditions. Both cotransport activity and intracellular ascorbic acid concentration increased in astrocytes activated by dibutyryl cyclic AMP. Conversely transport activity and ascorbic acid concentration were decreased by hyposmotic cell swelling, low extracellular Na(+) concentration, and depolarizing levels of extracellular K(+). In cells incubated for up to 3 h in medium having an ascorbic acid concentration typical of brain extracellular fluid, the changes in intracellular ascorbic acid concentration actually measured were not significantly different from those predicted by modeling changes in Na(+)-ascorbate cotransport activity. Thus, it was not necessary to specify alterations in vitamin C metabolism or efflux pathways in order to predict the steady state intracellular ascorbic acid concentration. These results establish that SVCT2 regulates intracellular ascorbic acid concentration in primary astrocyte cultures. They further indicate that the intracellular-to-extracellular ratio of ascorbic acid concentration at steady state depends on the electrochemical gradients of Na(+) and ascorbate across the plasma membrane. PMID- 11036153 TI - Convulsive seizures induced by N-methyl-D-aspartate microinjection into the mesencephalic reticular formation in rats. AB - Effects of microinjections of a single 2 or 10 nmol dose of N-methyl-D-aspartate (NMDA) into the unilateral mesencephalic reticular formation (MRF) on behavior and electroencephalogram were examined in rats (n=18) during a 15 min period (Exp. 1), and subsequent effects of sound stimulation with key jingling applied at 15, 30, and 45 min after the injections were observed (Exp. 2). The microinjections of 2 nmol dose of NMDA (n=10) induced hyperactivity (9 of 10 rats) and running/circling (8 of 10 rats) in Exp. 1, and hyperactivity (3 of 10 rats) in Exp. 2. Moreover, the microinjections of 10 nmol dose of NMDA (n=8) induced not only hyperactivity (8 of 8 rats) and running/circling (7 of 8 rats) but also generalized tonic-clonic seizures (GTCS) (5 of 8 rats) in Exp. 1; these seizure patterns were also elicited by sound stimulation in Exp. 2. The seizure patterns were accompanied by electroencephalographic seizure discharges in the MRF and the motor cortex. In contrast, the control group rats (n=10) which received a single dose of saline microinjection into the unilateral MRF showed no behavioral or electroencephalographic changes in both Exp. 1 and 2. These findings suggest that the MRF has an important role in the development of GTCS, which follows hyperactivity and running/circling, and that potentiation of excitatory neurotransmission in the MRF participates in the development of audiogenic seizures as well as GTCS. PMID- 11036154 TI - Are autonomic signals influencing cortico-spinal motor excitability? A study with transcranial magnetic stimulation. AB - In order to investigate the role of visceral afferent inputs flowing along autonomic fibers on corticospinal tract excitability, the variability of Motor Evoked Potentials (MEPs), elicited by Transcranial Magnetic Stimulation (TMS), was analysed during simultaneous monitoring of electrocardiogram (EKG) phases, breathing phases and sudomotor skin responses (SSRs) in a group of 10 healthy subjects. A cascade of at least 60 consecutive magnetic stimuli, with an interstimulus interval randomly varying between 20 and 40 s, was acquired. At the end of the recording session, the subject was asked to make at random five not consecutive self-paced forced inspirations. TMS was carried out at an intensity 10% above motor threshold excitability via a circular coil placed over the motor area of the right hemisphere. MEPs were recorded from the contralateral abductor digiti minimi muscle (ADM). Sudomotor Skin Responses (SSRs) were recorded on both hand palms. MEPs latency and amplitude did not show significant correlation with any of the EKG and respiratory phases. During forced inspiration, a significant latency shortening was found. TMS elicited SSRs, whose amplitudes were not correlated with MEP parameters. During forced inspiration a significant SSR amplitude increment, not correlated with MEP latency shortening, was also observed. These results assign a minor if any role to the considered autonomic parameters in modulating corticospinal motor excitability. PMID- 11036155 TI - Ca(2+)-independent activity of Ca(2+)/calmodulin-dependent protein kinase II involved in stimulation of neurite outgrowth in neuroblastoma cells. AB - We investigated the involvement of Ca(2+)-independent activity of Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) in stimulation of neurite outgrowth. When neuroblastoma Neruo2a (Nb2a) cells expressing the alpha isoform of CaM kinase II (Nb2a/alpha cells) were stimulated by plating, they changed shape from round to flattened, and began to form neurites within 15 min. Numbers of cells bearing neurites increased from 15 min to about 2 h. Neurite length increased markedly from 30 min to 2 h after stimulation. Ca(2+) independent activity of CaM kinase II increased immediately after stimulation, peaked at about 30 min, and then gradually decreased. Autophosphorylation of Thr 286 followed the same time course as the increase in Ca(2+)-independent activity. The autophosphorylation and appearance of Ca(2+)-independent activity preceded the formation of neurites. The effect of mutation of the autophosphorylation site in the kinase whose Thr-286 was replaced with Ala (alphaT286A kinase) or Asp (alphaT286D kinase) was examined. alphaT286A kinase was not converted to a Ca(2+) independent form, and alphaT286D kinase had Ca(2+)-independent activity significantly as an autophosphorylated kinase. Cells expressing alphaT286A kinase did not form neurites, and were indistinguishable from control Nb2a cells. Cells expressing alphaT286D kinase had much longer neurites than Nb2a/alpha cells expressing the wild type kinase, although the initiation of neurite outgrowth was very late. These results indicated that Ca(2+)-independent activity of the kinase autophosphorylated at Thr-286 involves for neurite outgrowth. PMID- 11036156 TI - Effects of subtypes alpha- and beta-adrenoceptors of the lateral hypothalamus on the water and sodium intake induced by angiotensin II injected into the subfornical organ. AB - The present experiments were conducted to investigate the role of the alpha(1A)-, alpha(1B), beta(1)- and beta(2)-adrenoceptors of the lateral hypothalamus (LH) on the water and salt intake responses elicited by subfornical organ (SFO) injection of angiotensin II (ANG II) in rats. 5-methylurapidil (an alpha(1A)-adrenergic antagonist), cyclazosin (an alpha(1B)-adrenergic antagonist) and ICI-118,551 (a beta(2)-adrenergic antagonist) injected into the LH produced a dose-dependent reduction, whereas efaroxan (an alpha(2)-antagonist) increased the water intake induced by administration of ANG II into the SFO. These data show that injection of 5-methylurapidil into the LH prior to ANG II into the SFO increased the water and sodium intake induced by the injection of ANG II. The present data also show that atenolol (a beta(1)-adrenergic antagonist), ICI-118,551, cyclazosin, or efaroxan injected into the LH reduced in a dose-dependent manner the water and sodium intake to angiotensinergic activation of SFO. Thus, the alpha(1)- and beta adrenoceptors of the LH are possibly involved with central mechanisms dependent on ANG II and SFO that control water and sodium intake. PMID- 11036157 TI - Prevention of 1-methyl-4-phenylpyridinium- and 6-hydroxydopamine-induced nitration of tyrosine hydroxylase and neurotoxicity by EUK-134, a superoxide dismutase and catalase mimetic, in cultured dopaminergic neurons. AB - Oxidative stress has been implicated in the selective degeneration of dopaminergic (DAergic) neurons in Parkinson's disease (PD). In this study, we tested the efficacy of EUK-134, a superoxide dismutase (SOD) and catalase mimetic, on the nitration of tyrosine hydroxylase (TH), a marker of oxidative stress, and neurotoxicity produced by 1-methyl-4-phenylpyridinium (MPP(+)) and 6 hydroxydopamine (6-OHDA) in primary DAergic neuron cultures. Exposure of cultures to 10 microM MPP(+) reduced dopamine (DA) uptake and the number of tyrosine hydroxylase immunoreactive (THir) neurons to 56 and 52% of control, while exposure to 30 microM 6-OHDA reduced DA uptake and the number of THir neurons to 58 and 59% of control, respectively. Pretreatment of cultures with 0.5 microM EUK 134 completely protected DAergic neurons against MPP(+)- and 6-OHDA-induced neurotoxicity. Exposure of primary neuron cultures to either MPP(+) or 6-OHDA produced nitration of tyrosine residues in TH. Pretreatment of cultures with 0.5 microM EUK-134 completely prevented MPP(+)- or 6-OHDA-induced nitration of tyrosine residues in TH. Taken together, these results support the idea that reactive oxygen species (ROS) are critically involved in MPP(+)- and 6-OHDA induced neurotoxicity and suggest a potential therapeutic role for synthetic catalytic scavengers of ROS, such as EUK-134, in the treatment of PD. PMID- 11036158 TI - Block of sodium currents in rat dorsal root ganglion neurons by diphenhydramine. AB - To elucidate the local anesthetic mechanism of diphenhydramine, its effects on tetrodotoxin-sensitive (TTX-S) and tetrodotoxin-resistant (TTX-R) sodium currents in rat dorsal root ganglion (DRG) neurons were examined by the whole-cell voltage clamp method. Diphenhydramine blocked TTX-S and TTX-R sodium currents with K(d) values of 48 and 86 microM, respectively, at a holding potential of -80 mV. It shifted the conductance-voltage curve for TTX-S sodium currents in the depolarizing direction but had little effect on that for TTX-R sodium currents. Diphenhydramine caused a shift of the steady-state inactivation curve for both types of sodium currents in the hyperpolarizing direction. The time-dependent inactivation became faster and the recovery from the inactivation was slowed by diphenhydramine in both types of sodium currents. Diphenhydramine produced a profound use-dependent block when the cells were repeatedly stimulated with high frequency depolarizing pulses. The use-dependent block was more pronounced in TTX R sodium currents. The results show that diphenhydramine blocks sodium channels of sensory neurons similarly to local anesthetics. PMID- 11036159 TI - Direct radiolabeling by [3H]quisqualic acid of group I metabotropic glutamate receptor in rat brain synaptic membranes. AB - [3H]Quisqualic acid (QA) was synthesized and used to label metabotropic glutamate receptor (mGluR) in rat brain synaptic membranes in the presence of three different ionotropic glutamate receptor agonists at respective saturating concentrations. Of several mGluR agonists tested, group I agonists were more potent in displacing [3H]QA binding than group II and group III agonists in the presence of the three ionotropic agonists. [3H]QA binding was markedly inhibited by guanine nucleotide analogues in a concentration-dependent manner at a concentration range of 10 nM to 1 mM. Scatchard analysis revealed that [3H]QA binding consisted of a single component with a K(d) of 50.9+/-5.3 nM and a B(max) of 431. 6+/-18.3 fmol/mg protein. These results suggest that [3H]QA indeed labels group I mGluR functionally coupled to GTP binding protein in rat brain synaptic membranes when determined under the experimental conditions employed. PMID- 11036160 TI - Antalarmin blockade of corticotropin releasing hormone-induced hypertension in rats. AB - Central administration of CRH results in endocrinological, cardiovascular, and behavioral effects that suggest stress or anxiety. Among these is a marked pressor response. Parenteral administration of CRH, however, results in hypotension. We used parenteral administration of antalarmin, a novel, small molecule CRH1 receptor antagonist, and alpha-helical CRH(9-41), a peptidic CRHR1/CRHR2 antagonist to attempt to determine the receptor mechanisms through which CRH is acting in both of these situations. Our results suggest that the hypertension produced by central CRH administration is mediated through central CRHR1 receptors, whereas the hypotension produced by parenteral CRH administration is mediated through peripheral CRHR2 receptors. PMID- 11036161 TI - Noncompetitive inhibition of the glycine receptor-mediated current by melatonin in cultured neurons. AB - The effect of melatonin on the glycine receptor-mediated response was studied in cultured chick spinal cord neurons using the whole-cell voltage-clamp recording technique. Melatonin rapidly and reversibly inhibited the glycine-induced current in a dose-dependent fashion, with an EC(50) of 934 microM and a maximal inhibition of 100%. Furthermore, melatonin noncompetitively inhibited the glycine response by an agonist-independent mechanism that was distinct from that of an open-channel blocker. PMID- 11036163 TI - Pentobarbital-induced modulation of flexor and H-reflexes in spinal rats. AB - Electrophysiological recordings of the H-reflex and nonnociceptive flexion reflex were obtained from pentobarbital-anesthetized Intact rats and from both, anesthetized and unanesthetized groups of Acute and Chronic Spinal rats. Results showed that the flexor, but not H-reflex, of Chronic Spinal rats was significantly larger than that of all other groups, which did not differ among themselves. The antispastic drug baclofen dose-dependently decreased the flexion response of Chronic Spinal rats (A(50)=4.3 mg/kg+/-2.1 and 9.0 mg/kg). PMID- 11036162 TI - Na(+)-Ca(2+) exchanger isoforms in rat neuronal preparations: different changes in their expression during postnatal development. AB - We examined the relative amounts of Na(+)-Ca(2+) exchanger (NCX) isoform mRNAs in cultured neurons, astrocytes and developmental rat brain. NCX1 transcript was predominant in neurons and astrocytes, but NCX2 transcript was about four-fold higher than NCX1 or NCX3 transcript in adult rat cortex. NCX2 transcript in the cortex increased markedly during postnatal development, whereas NCX1 and NCX3 transcripts decreased. Na(+)-dependent 45Ca(2+) uptake in the cortical homogenate increased significantly during postnatal development. PMID- 11036164 TI - Different astroglial reaction between the vagal dorsal motor nucleus and nucleus ambiguus following vagal-hypoglossal nerve anastomosis in cats. AB - The dorsal motor nucleus of the vagus (DMV) and nucleus ambiguus (NA) were both traced with horseradish peroxidase (HRP) retrograde labelling technique after vagal-hypoglossal nerve anastomosis (VHA). By light microscopy, reinnervation of the new target, viz. tongue skeletal musculature, by DMV and NA was established at 22 days postoperation (dpo) as shown by the neuronal labelling with HRP. Ultrastructurally, signs of retrograde degeneration occurred in some DMV and NA neurons between 3 and 25 days after VHA. The incidence of darkened dendrites, an early sign of dendritic loss, was more common in the DMV compared to the NA. Accompanying the neuronal alteration were drastic astrocytic reactions in the DMV, but not in the NA. Between 3 and 7 dpo, the astrocytes in the DMV showed extensively hypertrophied processes and by 22 dpo, the somata and dendrites of HRP-labelled DMV neurons, but not NA's, appeared to be delineated by the increased lamellar astrocytic processes. Such a feature was sustained throughout the remaining postoperative intervals up to 500 dpo. It is concluded that the DMV motoneurons being autonomic in nature are probably not conducive to the newly acquired target organ. Hence, the insulation of the regenerating DMV motoneurons by the astroglial ensheathment would be vital in the neuronal remodelling and reconstruction of the vagal-hypoglossal pathway. PMID- 11036165 TI - Interferon beta-1a downregulates TNFalpha-induced intercellular adhesion molecule 1 expression on brain microvascular endothelial cells through a tyrosine kinase dependent pathway. AB - TNFalpha (100 U/ml, 24 h) upregulated intercellular adhesion molecule 1 (ICAM1) expression on brain microvascular endothelial cell (BMEC) culture. The tyrosine kinase (TK) inhibitor genestein (100 microgram/ml), the protein kinase C (PKC) inhibitor staurosporin (1 nM), and interferon (IF) beta-1a (1000 U/ml) antagonized TNFalpha effect. When an ineffective dose of IFbeta-1a (100 U/ml) was challenged with ineffective doses of either genestein (10 microgram/ml) or staurosporin (0.1 nM), the combination IFbeta-1a-genestein significantly reduced TNFalpha-induced ICAM1 expression whereas IFbeta-1a-staurosporin did not. These findings indicate that a TK- rather than a PKC-dependent mechanism is involved in the modulation of TNFalpha response by IFbeta-1a on BMECs. PMID- 11036166 TI - Nitric oxide is involved in sustained and delayed cell death of rat retina following transient ischemia. AB - We have investigated the role of nitric oxide (NO) in the rat retina following ischemic injury induced by transient increase of intraocular pressure. The thickness of both the inner plexiform layer and inner nuclear layer decreased during early postischemic stages (up to 1 week). In late postischemic stages (2-4 weeks), the thickness of the outer nuclear layer (ONL) decreased markedly. Thus, mechanisms other than excitotoxic ones may contribute to postischemic retinal cell death. Treatment of rats with N(G)-nitro-L-arginine methyl ester, a nitric oxide synthase (NOS) inhibitor, significantly reduced ischemic damage. Our findings suggest that NO is involved in the mechanism of ischemic injury, and plays a key role in the delayed and sustained cell death in the ONL following transient retinal ischemia. PMID- 11036167 TI - Anesthetic concentrations of riluzole inhibit neuronal nitric oxide synthase activity, but not expression, in the rat hippocampus. AB - We hypothesized that anesthetic dose of riluzole, an inhibitor of glutamate neurotransmission, may affect the activity and/or expression of neuronal NOS (nNOS). Riluzole, N(G)-nitro-L-arginine-methyl ester (L-NAME) and 7-nitro indazole (7-NI) produced a concentration-related inhibition of nNOS activity in vitro. Riluzole competed with 7-NI for inhibition of nNOS activity, but had no effect on nNOS or endothelial NOS (eNOS) protein expression. Also, nNOS activity was significantly decreased in riluzole-anesthetized rats (40 mg kg(-1) i.p., 32+/-6% from controls, P<0.05). Therefore, blockade of nNOS activity may be involved in the anesthetic effects of riluzole in vivo. PMID- 11036168 TI - Plasma cortisol levels in elderly female subjects with Alzheimer's disease: a cross-sectional and longitudinal study. AB - We investigated the plasma cortisol levels at a fasting state in elderly female Alzheimer's disease (AD), vascular dementia (VD), and non-demented subjects (n=66, 28 and 21, respectively). Twenty-eight AD subjects were followed for 40 months. The plasma cortisol levels in AD and VD subjects were significantly higher than those of non-demented subjects at baseline. In AD subjects in relatively early stages of the disease [Mini-Mental State Examination (MMSE)], at baseline, high plasma cortisol led to rapid declines in MMSE scores over a 40 month period. PMID- 11036169 TI - Projections from the rostral pole of the inferior colliculus to the cat superior colliculus. AB - The neuroanatomical data given here reveal a dense projection from the rostral pole of the cat inferior colliculus (rpIC) to the superior colliculus (SC). A portion of this pathway distributes in 'patches' across the ventral portion of the intermediate grey layer. These finding suggest that the rpIC input to the SC might play a role in determining the auditory receptive fields of SGI neurons and in the construction of the SC's precise two dimensional map of auditory space. PMID- 11036170 TI - Erratum to: effects of blockade of voltage-sensitive Ca(2+)/Na(+) channels by a novel phenylpyrimidine derivative, NS-7, on CREB phosphorylation in focal cerebral ischemia in the rat PMID- 11036171 TI - Neuromodulations of the intractable neurologic symptoms PMID- 11036172 TI - Neuromodulation: an overview. AB - For over two centuries, electricity has been known to induce modification of neural and nerve fiber activity and has been proposed to be used to treat some neurological dysfunctions. The new era of the use of electrical current in the treatment of neurological symptoms began in 1967 with the use of totally implanted devices that deliver a controlled amount of electricity on a precise structure within the nervous systems and was first used to control pain. Extensive research has been carried out ever since to elucidate the mechanism of action of this treatment and extend its indication for the treatment of the other neurological symptoms. So far, there is evidence that the treatment is safe and efficient for long periods of time, as it does not induce permanent damage to the stimulated structure. Most likely, electrical current at the parameters used for therapeutic purpose induces an inhibition of the structure on which it is applied. However, this may be accompanied by either inhibition or excitation of anatomically related structures. For this reason, it seems more convenient to refer to this type of therapy as neuromodulation.A review of the historical development of this fascinating area is presented, with special attention to the evidence derived from experimental work on the parameters that electrical current must maintain to avoid damage to the underlying tissue. PMID- 11036173 TI - Instrumentation for neuromodulation. AB - The explosion in understanding how the central nervous system (CNS) works affords new opportunities to interact with the nervous system to compensate for dysfunction due to disease or injury. Neuromodulation is a term that describes methods that carry out that interaction based on principles of nerve cell physiology. There currently are two neuromodulation techniques used that require implantable devices-neurostimulation and implantable, chronic drug delivery. This article describes the devices used for neuromodulation, the motivation for the different feature sets of the devices, and the physiological and technological principles underpinning their use. PMID- 11036174 TI - Functional imaging and neurophysiological assessment of spinal and brain therapeutic modulation in humans. AB - We summarize here our experience in the neurophysiological and neuroimaging assessment of spinal and brain neuromodulation for pain relief. Techniques reviewed include somatosensory evoked potentials (SEPs), nociceptive spinal (RIII) reflexes, and positron emission tomography (PET), which have been applied both to investigate the mechanisms and to optimize the application of neurostimulation procedures. SEPs are especially useful in the preoperative assessment of patients with neuropathic pain, as they allow the establishment of the functional state of the dorsal column system. Patients with strongly abnormal SEPs due to ganglionic or preganglionic pathology are not likely to benefit from spinal (SCS) or peripheral (TENS) neurostimulation, because ascending fibers disconnected from their soma will undergo rapid degeneration and not be excitable. In the postoperative period, nociceptive spinal reflexes yield objective data concerning the effects of neurostimulation on spinal circuitry. In our experience, the best clinical results are achieved in patients with preserved preoperative SEPs, in whom neurostimulation entails profound attenuation of nociceptive reflexes.PET-scan imaging techniques have recently been used to demonstrate changes in cerebral blood flow during new neuromodulation schemes such as motor cortex stimulation for pain control (MCS). PET studies highlight the thalamus as the key structure mediating functional MCS effects. Thalamic activation would trigger a cascade of synaptic events influencing activity in other pain-related structures including the anterior cingulate gyrus, insula, and upper brainstem. The combination of clinical electrophysiology and functional neuroimaging provides insight into the mechanisms of action of neuromodulation procedures, guides clinical decision, and contributes to optimize patient selection. PMID- 11036175 TI - Spinal cord stimulation for chronic pain management. AB - This article outlines the role of spinal cord stimulation in contemporary chronic pain management. The anatomical and neurophysiological correlates of stimulation of the intraspinal structures are discussed. The most common indications are presented, including failed back syndrome, reflex sympathetic dystrophy, neurogenic thoracic outlet syndrome, and spinal cord injury, etc. The most common complications are presented, including paralysis, infection, electrode migration, cerebrospinal fluid leak, and pain. Spinal cord stimulation is one of the most effective techniques available in the management of severe chronic pain that has been refractory to other more conservative modalities. PMID- 11036176 TI - Motor cortex stimulation in the treatment of central and neuropathic pain. AB - BACKGROUND: Motor cortex stimulation has been proposed for the treatment of central pain. METHODS: Thirty-two patients with refractory central and neuropathic pain of peripheral origin were treated by chronic stimulation of the motor cortex between May 1993 and January 1997. The mean follow-up was 27.3 months. The first 24 patients were operated on according to the technique described by Tsubokawa. The last 13 cases (8 new patients and 5 reinterventions) were operated on by a technique including localization by superficial CT reconstruction of the central region and neuronavigator guidance. The position of the central sulcus was confirmed by the use of intraoperative somatosensory evoked potentials. The somatotopic organization of the motor cortex was established preoperatively by studying the motor responses at stimulation of the motor cortex through the dura. RESULTS: Ten of the 13 patients with central pain (77%) and 10 of the 12 patients with neuropathic facial pain experienced substantial pain relief (83.3%). One of the three patients with post-paraplegia pain was clearly improved. A satisfactory result was obtained in one patient with pain related to plexus avulsion and in one patient with pain related to intercostal herpes zoster. None of the patients developed epileptic seizures. CONCLUSIONS: Our results confirm that chronic stimulation of the motor cortex is an effective method in treating certain forms of refractory pain. PMID- 11036177 TI - Vim thalamic stimulation for tremor. AB - Chronic stimulation of the ventral intermediate nucleus (Vim) of the thalamus is highly effective for the treatment of tremor. Patients with tremor associated with Parkinson's disease and essential tremor appear to respond best. Patients with cerebellar tremors may also respond but to a lesser extent. Although tremor is improved, Vim DBS does not substantially improve the daily living activities of patients with Parkinson's disease. This is related to the lack of effect on rigidity, bradykinesia, and gait and postural disturbances associated with Parkinson's. For this reason, the majority of patients with Parkinson's disease who require surgery are better treated with interventions in the globus pallidus or subthalamic nucleus, targets that allow improvement in all cardinal features of Parkinson's disease. In contrast, Vim DBS has unequivocal functional benefit in patients with essential tremor, this is likely to remain the major indication of this form of therapy. The mechanism of action of thalamic DBS is not understood and remains a research priority. PMID- 11036178 TI - Subthalamic prelemniscal radiation stimulation for the treatment of Parkinson's disease: electrophysiological characterization of the area. AB - Previous reports have provided evidence of a reticulo-thalamic system, extending from the mesencephalic reticular formation (MRF) to the ventrolateral thalamus (VL), involved in the production of tremor. In humans, a funnel of fibers in the posterior subthalamus named the prelemniscal radiations (Raprl) has been described as an exquisite target to treat tremor in cases of Parkinson's disease. In the present study, a group of 14 patients suffering from Parkinson's disease, with prominent unilateral tremor and rigidity, were implanted with tetrapolar depth brain stimulation (DBS) electrodes in Raprl to perform chronic electrical stimulation (ES) for the treatment of patient symptoms. Electrodes were left externalized to corroborate their placement throughout MRI studies and also to perform the following electrophysiological battery: (a) recording of somatosensory-evoked responses (SEP) through different electrode contacts and scalp by means of a paradigm to study the attention process; (b) evoking scalp EEG responses by stimulation with low (3 cps, 6 cps) and high (60-120 cps) frequencies with stimuli delivered through different electrode contacts, and (c) studying recovery cycle (RC) potentials in the Raprl while the upper MRF was being stimulated and, conversely, the RC in MRF while Raprl was being stimulated, before and after subacute Raprl stimulation. Thereafter, the electrodes were internalized and connected to a pulse generator (IPG) to carry on chronic ES, while the effects of stimulation were determined through a quantitative evaluation that measured phasic and tonic muscular activity with EMG recordings during different motor tasks. Results indicate the following: (a) that late, but not early, SEP components were recorded in Raprl and modulated in different attentive conditions; (b) that bilateral recruiting responses and spike and wave complexes were elicited by Raprl through low-frequency stimulation, while bilateral positive DC shifts induced by high-frequency stimulation were recorded, similar to those obtained in animals from MRF, and (c) that Raprl-ES induced RC inhibition at Raprl, but Raprl ES did not change MRF-RC. Long-term Raprl-ES induced a significant decrease in tremor and rigidity. It was concluded that Raprl represents a subthalamic circuit electrophysiologically related to MRF in the genesis of tremor and rigidity and in the process of selective attention. Raprl-ES induced a significant improvement in tremor and rigidity by causing inhibition of the stimulated area. PMID- 11036180 TI - Cerebellar stimulation for cerebral palsy spasticity, function, and seizures. AB - Chronic cerebellar stimulation (CCS) applied to the superio-medial cortex reduces generalized cerebral spasticity, athetoid movements, and seizures. Eighteen clinics have reported on 600 cerebral palsy (CP) patients who comprise 90% of those treated with CCS. CP patients have varying degrees of limited abilities interfered with by spasticity (primitive reflexes, increased muscle tone, co contractions, and spasms) and by athetoid movements in two-thirds of the patients. With CCS, spasticity reduction occurred in 85% (marked 25%, moderate 34%, mild 27%) and resulted in improvements in patient drooling, speech, respiration, posture, motor performance, gait, joint range of motion, and mood states. Radiofrequency (RF)-linked stimulators were used initially with serious equipment and calibration problems; 68% of 422 patients improved. When totally implantable controlled-currrent stimulators were used, 86% of 178 patients improved. Our double-blind study of 20 CP patients using this implantable stimulator showed 12 (60%) improved in motor performance, joint range of motion, and profile of mood states when the stimulator was ON. When abilities are graded (1: poor to 9: best), the seven patients with the higher functioning grades (5-8) all improved (99% confidence level). Intractable seizures occurred in 27 (8%) of our CP patients. At a 17-year follow-up, 19 patients contacted were using or had used CCS with 10 (53%) seizure-free and 6 (32%) with reduced seizures. CCS should be given by a totally implanted controlled-current stimulator (1-4 microCoulombs/sq. cm. /phase, 150-200 Hz) applied intermittently to the superio medial cerebellar cortex for safe, effective, and continuous results. PMID- 11036179 TI - Subthalamic stimulation for Parkinson's disease. AB - Deep brain stimulation by high frequency (HFS) has been developed starting in the thalamic target (Vim) from pragmatic observations and subsequently followed by other targets, such as the subthalamic nucleus (STN) and pallidum as an application of current knowledge from basic preclinical research in neuroscience. The mechanism involved by this neurosurgical approach is not completely solved. For Vim we have formed the hypothesis that HFS induces a jamming of sensory-motor loops but for the STN, from our experimental research in rats we have shown that HFS induces functional inhibition of cell activity in the target nuclei. In our patients the implantation of the stimulation electrodes was carried out stereotactically, under local anesthesia, using ventriculography, MRI, microrecordings and clinical evaluation of the effects of stimulation on rigidity. When the stimulation is turned ON in the STN area a significant decrease in rigidity was determined by the neurologists. Stimulation or even penetration of the electrode may be responsible for transient dyskinesias. The average location of the clinically efficient contact of the chronic stimulating electrodes is statistically located at 5.02 +/- 0.71 1/12 degrees of AC-PC in the AP direction, at -1.5 +/- 0.66 1/8 degrees of the height of the thalamus in the ventricle direction, with laterality at 11.98 +/- 1.12 mm in the lateral direction. The beneficial effects of STN stimulation are significant providing that the electrodes are correctly placed into the target. There is strong improvement of the symptoms of the triad in which akinesia, rigidity, and tremor are reduced on average to 41. 6, 48.6, and 27%, respectively, when compared with the previous preoperative level. From our experience, HFS of the STN could be considered the surgical therapy of choice at advanced stages of Parkinson's disease. PMID- 11036181 TI - Vagus nerve stimulation for seizures. AB - It is agreed that 1% of the general population is afflicted with epilepsy and close to 30% of epilepsy patients are intractable to medications. In spite of a recent increase in the number of new medications that are available on the market, many patients continue to have seizures or their seizures are controlled at the expense of intolerable side effects. Resection epilepsy surgery is an alternative; however, not every intractable patient is a good candidate for this surgery. Additionally, it is only offered to a small fraction of these patients due to the lack of an adequate number of comprehensive epilepsy programs and financial support for such surgeries. Vagus nerve stimulation (VNS) is a novel adjunctive therapy that has recently become commercially available for intractable epilepsy. It is indicated as an add-on treatment for seizures of partial onset with or without secondary generalization in patients 12 years of age or older. The VNS system is comprised of a battery generator that delivers regular intermittent electrical stimuli programmed via menu-driven software and an interrogating wand. The generator is implanted in the left upper chest and connected to the left cervical vagus nerve via a pair of semi-circular helical electrodes wound around the vagus nerve and wires tunneled under the skin. Surgery is normally completed within 2 h under general anesthesia and the patient can go home within a few hours postoperatively. Experiments in humans began in 1988 with two single-blind pilot studies that demonstrated the feasibility and safety of this unconventional therapy. Following these studies, two multicenter, active-control, parallel, double-blind protocols showed a statistically significant reduction in partial onset seizures with reasonable and well tolerated side effects. Adverse events related to VNS included voice alteration and a tingling sensation in the throat during stimulation only and a decrease in intensity over several weeks. Coughing during stimulation occurred normally when therapy was initiated and shortness of breath occurred mainly during exertion. Long-term follow-up suggests that reduction in seizure frequency and intensity is maintained over time. VNS is a novel adjunctive anti-epilepsy therapy that offers patients a better-tolerated option than medications in general and that is less invasive and extensive than resection surgery. Its efficacy may compare to novel potent anti-epilepsy drugs; however, VNS does not replace resection epilepsy surgery in selected patients in whom chances of seizure-free results are high (70 90%). PMID- 11036182 TI - Acute and chronic electrical stimulation of the centromedian thalamic nucleus: modulation of reticulo-cortical systems and predictor factors for generalized seizure control. AB - The present report recapitulates the clinical and electrophysiologic studies we have performed on patients with certain forms of medically intractable epilepsy to investigate the basic mechanisms and predictor factors for seizure control of the electrical stimulation of the thalamic centromedian nucleus (CM) procedure. Acute electrical stimulation of CM reveals that in humans, as in other animals, CM represents a thalamic relay of a reticulo-cortical system that participates crucially in wakefulness and attentive processes and in regulation of cortical excitability, as well as in the physiopathology of genuine generalized epileptic seizures. For example, unilateral, threshold, low-frequency (6/sec) stimulation of CM produced electrocortical incremental responses, while high-frequency (60/sec) stimulation of CM produced electroencephalogram (EEG) desynchronization and electronegative DC shifts with no behavioral counterparts. In contrast, combined suprathreshold low-frequency (3/sec) stimulation of CM on one side and of mesencephalic reticular stimulation on the other produced generalized spike wave complex discharges accompanied by the symptoms of a typical absence attack, including motionless stare, eye blinking, and unresponsiveness of patients to a series of flashes under a simple response task. Chronic bilateral, threshold, high-frequency (60/sec) stimulation of CM significantly decreased the number of primary and secondary generalized tonic-clonic seizures and atypical absence attacks and the amount of interictal generalized EEG discharges in both. In addition, it improved the psychological performance of patients and normalized the EEG by increasing the frequency of background EEG activity. In contrast, chronic stimulation of CM reduced neither the number of complex partial seizures nor the epileptic EEG activities localized in the temporal region. Good outcomes of the chronic CM stimulation procedure were achieved depending on correct selection of patients and accuracy of ventriculographic stereotactic targets, as well as on periodic clinical and EEG evaluation and electrophysiologic monitoring of CM electrical stimulation reliability. However, the presence of 3- to 6-month long-lasting effects of CM stimulation made statistical evaluation of ON-OFF effects of CM stimulation under placebo, double-masked randomized experiments difficult. PMID- 11036183 TI - Subacute and chronic electrical stimulation of the hippocampus on intractable temporal lobe seizures: preliminary report. AB - Recent animal experiments show that the application of an electrical stimulus to the amygdala or hippocampus following the kindling stimulus produced a significant and long-lasting suppressive effect on this experimental model of epilepsy. This is a preliminary report on the development of a surgical neuromodulatory procedure by chronic electrical stimulation of the hippocampus (CHCS) for control of intractable temporal lobe seizures in patients in whom anterior temporal lobectomy is not advisable, i.e., patients with bilateral temporal foci or a unilateral focus spreading to surrounding cerebral regions of the dominant hemisphere. This work was divided in two main consecutive stages. In the first stage, we demonstrated that subacute hippocampal stimulation (SAHCS) blocks intractable temporal lobe epileptogenesis with no additional damage to the stimulated tissue, and in a second stage, we attempt to demonstrate that CHCS may produce a sustained, long-lasting antiepileptic condition without additional undesirable effects on language and memory. In addition, taking advantage of this unique and ethically permissible situation, we attempt to determine whether or not the antiepileptic effects of SAHCS and CHCS are due to inhibition of the stimulation of hippocampal tissue by means of a number of electrophysiological, single photon computed tomography (SPECT) perfusion, and autoradiographic techniques.SAHCS during 3-4 weeks prior to anterior temporal lobectomy applied to a critical area located either at the anterior Pes hippocampus close to the amygdala or at the parahippocampal gyrus close to the entorhinal cortex abolished clinical seizures and significantly decreased the number of interictal spikes at focus after 5-6 days. Microscopy analysis of the stimulated tissue showed no evident histopathological differences between stimulated vs. non-stimulated hippocampal tissues. Additionally, CHCS persistently blocked temporal lobe epileptogenesis for 3-4 months with no apparent additional undesirable effects on short memory. Also, inhibition of the stimulated hippocampus seems to be one of the possible mechanisms underlying the beneficial antiepileptic effects of SAHCS and CHCS. This was revealed by increased threshold and decreased duration of the afterdischarges induced by hippocampal stimulation, flattening of the hippocampal evoked response recovery cycles, SPECT hypoperfusion of the hippocampal region, and increased hippocampal benzodiazepine receptor binding. Future studies increasing the number and time of follow-up of patients under hippocampal stimulation are necessary before considering CHCS a reliable procedure for controlling intractable temporal lobe seizures. PMID- 11036184 TI - Allosteric modulation in spontaneously active mutant gamma-aminobutyric acid(A) receptors [corrected]. AB - Tryptophan substitutions were made in the second transmembrane domain of the gamma-aminobutyric acid(A) (GABA(A)) receptor alpha and beta subunits and the resulting mutant receptors, containing alpha(2)(S270W) and/or beta(1)(S265W), were expressed in Xenopus oocytes. Mutation of either or both subunits resulted in receptors that exhibited enhanced sensitivity to agonist and were spontaneously active in the absence of GABA. The spontaneous activity was blocked by picrotoxin or bicuculline. The enhancement of GABA-induced currents by pentobarbital, by the neurosteroid 5alpha-pregnan-3alpha-ol-20-one, and by the benzodiazepine flunitrazepam was dramatically reduced in the mutant receptors. These results are consistent with the idea that a mutation that promotes gating behavior in a ligand-gated ion channel will also show reduced effects of all positive allosteric modulators in a generalized manner, even when these modulators act at distinct sites on the receptor. PMID- 11036186 TI - Attenuated Creutzfeldt-Jakob Disease agents can hide more virulent infections. AB - We previously showed that a slow infectious strain of Creutzfeldt-Jakob Disease (CJD) can dramatically suppress the expression of a fast virulent agent injected intracerebrally 80days later. While the slow SY agent eventually produced disease at approximately 400days, there was little evidence of the fast FU agent. However, two of 18superinfected mice showed a minor increase in pathologic changes. To determine if FU was partially or completely suppressed, or if FU and SY agents formed a 'chimera' with intermediate incubation properties as predicted by prion theory, we passaged representative brains. All traces of FU were obliterated in typical brains of suppressed mice. The two aberrant mice however had mixed SY and FU infections, with FU reappearing at late stages of SY disease. Thus less virulent sporadic CJD infections in older people can conceal other agents such as variant CJD, the more recently evolved and virulent agent linked to bovine spongiform encephalopathy. This powerful model of agent-induced repression also implicates targets other than prion protein (PrP) in eliminating infection. PMID- 11036185 TI - Prolonged exposure to hyperbaric oxygen induces neuronal damage in primary rat cortical cultures. AB - While seizure attack is one of the serious complications during the hyperbaric oxygen (HBO) therapy, there is still no direct evidence showing that HBO can induce neuronal damage in the brain. The objective of this study was first to investigate whether HBO would lead to neurotoxicity in the primary rat cortical culture. Second, since alterations in neurotransmitters have been suggested in the pathophysiology of central nervous system (CNS) oxygen toxicity, the protective effects of the N-methyl-D-aspartate (NMDA) receptor antagonism and nitric oxide (NO) synthase inhibition on the HBO-induced neuronal damage were examined. The results showed that HBO exposure to 6 atmosphere absolute pressure (ATA) for 30, 60, and 90 min increased the lactate dehydrogenase (LDH) activity in the culture medium in a time-dependent manner. Accordingly, the cell survival, measured by the 3,(4,5-dimethyl-2-thiazolyl)2, 5-diphenyl-tetrazolium bromide (MTT) assay, was decreased after HBO exposure. Pretreatment with the NMDA antagonist MK-801 protected the cells against the HBO-induced damage. The protective effect was also noted in the cells pretreated with L-N(G)-nitro arginine methyl ester, an NO synthase inhibitor. Thus, our results suggest that activation of NMDA receptors and production of NO play a role in the neurotoxicity produced by hyperbaric oxygen exposure. PMID- 11036187 TI - Direct visualization of cholecystokinin subtype2 receptors in rat central nervous system using anti-peptide antibodies. AB - The cholecystokinin receptor, subtype 2 (CCK(2)R), is considered, based on receptor autoradiography, to be the predominant receptor for this peptide transmitter in the mammalian central nervous system. To directly visualize the CCK(2)R we utilized a convenient and sensitive immunohistochemical procedure using antipeptide receptor antibodies raised in rabbits against unique portions of the carboxyl tail and third intracellular loop of the CCK(2)R. Antibodies were characterized by ELISA and Western blotting, and used for immunohistochemistry in rat brain sections. Studies with both antibodies revealed a widespread topographic distribution of CCK(2)R-like immunoreactivity (CCK(2)R-LI) in regions such as cortex, olfactory bulb, nucleus accumbens, septum, striatum, hippocampus, basolateral amygdala, habenula, hypothalamus, thalamus, ventral mesencephalon, inferior colliculus, parabrachial nucleus, pontine nucleus, supercolliculus, red nucleus, subcommisural and occulomotor nucleus, area postrema, solitary, olivary, cochlear, cuneate and trigeminal nuclei and spinal cord dorsal horn in agreement with the results of previous receptor autoradiography. PMID- 11036188 TI - Oxidative stress and an altered methionine metabolism in alcoholism. AB - The exact mechanism of brain atrophy in patients with chronic alcoholism remains unknown. There is growing evidence that chronic alcoholism is associated with oxidative stress and with a derangement in sulphur amino acid metabolism (e.g. ethanol-induced hyperhomocysteinemia). Furthermore, it has been reported that homocysteine induces neuronal cell death by stimulating N-methyl-D-aspartate receptors as well as by producing free radicals. To further evaluate this latter hypothesis we analysed serum levels of both homocysteine and markers of oxidative stress (malondialdehyde) in alcoholic patients who underwent withdrawal from alcohol. Homocysteine and malondialdehyde were quantified by high performance liquid chromatography (HPLC) in serum samples of 35 patients (active drinkers). There was a significant correlation (P<0. 01) between blood alcohol concentration and elevated homocysteine (Spearman's r=0.71) and malondialdehyde (r=0.90) levels on admission. In addition, homocysteine and malondialdehyde levels were found to be significant decreased after 3 days of withdrawal treatment (Wilcoxon test: homocysteine, Z=-5.127; malondialdehyde, Z=-3.120; P<0.01). We postulate that excitatory neurotransmitters and mechanisms of oxidative stress in patients with chronic alcoholism may partly mediate excitotoxic neuronal damage and hereby cause brain shrinkage. PMID- 11036189 TI - Feeding-induced c-fos expression in the nucleus of the solitary tract and dorsal medullary reticular formation in neonatal rats. AB - We investigated feeding-associated activation of neurons in the medulla oblongata during the suckling period in rats, using the c-fos gene-encoded protein (Fos) immunohistochemistry. After an isolation from mothers for 12 h, neonates were either breast-fed intensively or further isolated for another 3 h, and sacrificed on postnatal day 1, 3, 5, 7 and 14 (P1-14). In the former pups, Fos immunoreactive (FI) neurons were predominantly localized in the nucleus of the solitary tract (NST) and in the dorsal medullary reticular formation (RF). The number of FI cells peaked on P5-7 and decreased on P14 in the NST, and increased remarkably on P3 and was consistently high until P14 in the dorsal RF. In contrast, much fewer FI cells were found in the NST and RF in the latter pups. The results indicated that not only the NST but also the dorsal RF were implicated in feeding behavior in the suckled pups. PMID- 11036190 TI - Substance P modulation of striatal dopamine outflow is determined by M(1) and M(2) muscarinic receptors in male wistar rats. AB - Substance P (SP) stimulates striatal dopamine outflow through a cholinergic muscarinic link. SP-induced increase in acetylcholine (Ach) is concentration dependent, whereas the stimulation of dopamine outflow is seen only over a limited concentration range. M(1) and M(2) receptor stimulation has opposite effects on dopamine outflow. We postulated that the effect of SP on dopamine outflow depends on the M(1)/M(2) balance. We show that Ach (10-2500 microM) stimulates dopamine outflow in striatal slices in a biphasic manner, similar to SP (0.01-100 nM). An inactive SP concentration (10 nM) which was higher than the active concentration range, became active in the presence of the M(2) antagonist methoctramine (100 microM). Conversely, the effect of 1 nM SP was reversed by the M(1) antagonist pirenzepine (1 microM). Our observations show that SP modulation of dopamine outflow is determined by a balance between M(1) and M(2) receptors. PMID- 11036191 TI - Effects of alpha-dendrotoxin and dendrotoxin K on extracellular excitatory amino acids and on electroencephalograph spectral power in the hippocampus of anaesthetised rats. AB - Dendrotoxins, important pharmacological tools for studying K(+) channels, are potently convulsant in the central nervous system and evidence suggests that different members of the dendrotoxin family may act at pre- or post-synaptic sites. Using a combination of intrahippocampal infusion, microdialysis and electroencephalograph (EEG) recording, we have compared the effects of alpha dendrotoxin and dendrotoxin K on extracellular levels of excitatory amino acids in anaesthetised rats. Our findings show that although infusion of 35 pmol of both peptides was associated with elevated extracellular aspartate and glutamate, these increased levels were more sustained with dendrotoxin K. Furthermore, there was EEG evidence of an associated transient functional change consistent with an action on pre-synaptic K(+) channels. In contrast, infusion of alpha-dendrotoxin produced only a brief effect on amino acid levels and no evidence of a functional consequence. PMID- 11036192 TI - Middle and long latency peak sources in auditory evoked magnetic fields for tone bursts in humans. AB - The relative position of the P50m and the N100m sources of the auditory evoked magnetic field remains unclear. Magnetoencephalography was performed in 24 normal subjects. Contralateral P50m to left and right ear stimulus was observed in 21 and 19hemispheres, respectively. Ipsilateral P50m to left and right ear stimulus was observed in 17 and 16hemispheres, respectively. N100m was observed in all subjects for all stimuli. Relative position of the equivalent current dipole of the P50m was 1.0+/-7.6 (mean+/-SD) mm posterior, 2.0+/-5.8mm inferior and 1.8+/ 8.0mm medial to the N100m dipole position considering all observations. We suggest that the P50m and N100m sources are colocated in an extended area of the cortex. PMID- 11036193 TI - Differences in event-related and induced brain oscillations in the theta and alpha frequency bands related to human intelligence. AB - High (intelligence quotient (IQ)=126) and low intelligent individuals (IQ=88) were listening to tone pips and performed an auditory oddball task while their electroencephalogram was recorded. Significant differences relating to intelligence were observed in induced and event related band power in the theta (4-7 Hz) and upper alpha band (10-13 Hz). In the oddball task high intelligent individuals displayed less theta synchronization and more synchronization in the upper alpha band. The results were explained by the more efficient engagement of neural networks in more intelligent individuals. PMID- 11036194 TI - Cutaneous nerve degeneration induced by acrylamide in mice. AB - Acrylamide is a neurotoxin producing distal axonopathy. Previous studies mainly focused on large-diameter motor and sensory nerves, and the influences of acrylamide neurotoxicity on small-diameter sensory nerves in the skin remained elusive. We investigated skin innervation in mice intoxicated by acrylamide. Small-diameter sensory nerves in the skin degenerated after acrylamide intoxication. Epidermal nerve swelling was the earliest sign of acrylamide intoxication, with 29.5+/-2.4% of swollen epidermal nerves in the initial stage (P<0.001). There was a trend of progressive loss of epidermal nerves with a significantly reduced epidermal nerve density in the late stage (P<0.003). In the mean time, degenerating dermal nerves exhibited a beaded appearance. These results suggest the scenario of small-diameter cutaneous nerve degeneration in acrylamide neurotoxicity: beginning with epidermal nerve terminal swelling in the initial stage and resultant epidermal nerve depletion in the late phase. PMID- 11036195 TI - Calcium-dependent modulation by ethanol of mouse synaptosomal pyroglutamyl aminopeptidase activity under basal and K(+)-stimulated conditions. AB - We studied the in vitro effects of ethanol (25, 50 and 100 mM) on pyroglutamyl aminopeptidase activity (pGluAP), which has been reported as thyrotrophin releasing-hormone-degrading activity. pGluAP was measured in presence or absence of calcium, under basal and K(+)-stimulated conditions, in synaptosomes and their incubation supernatant, using pyroglutamyl-beta-naphthylamide as substrate. In basal conditions, in synaptosomes, pGluAP was inhibited by ethanol in a calcium independent way. In the supernatant, the response differed depending on the concentration of ethanol. Depolarization with K(+) modified pGluAP in synaptosomes and supernatant depending on the presence or not of calcium. In synaptosomes, in absence of calcium, the activity was inhibited at the highest concentrations of ethanol. In contrast, in the supernatant, under depolarizing conditions, ethanol increases pGluAP in absence of calcium. These changes may be in part responsible of the behavioural changes associated to alcohol intake. PMID- 11036196 TI - Dental occlusion modifies gaze and posture stabilization in human subjects. AB - Repercussion of dental occlusion was tested upon postural and gaze stabilization, the latter with a visuo-motor task evaluated by shooting performances. Eighteen permit holders shooters and 18 controls were enrolled in this study. Postural control was evaluated in both groups according to four mandibular positions imposed by interocclusal splints: (i) intercuspal occlusion (IO), (ii) centric relation (CR), (iii) physiological side lateral occlusion and (iv) controlateral occlusion, in order to appreciate the impact of the splints upon orthostatism. Postural control and gaze stabilization quality decreased, from the best to the worst, with splints in CR, IO and lateral occlusion. In shooters, the improvement in postural control was parallel to superior shooting performance. A repercussion of dental occlusion upon proprioception and visual stabilization is suggested by these data. PMID- 11036197 TI - Caspase-3 activation by beta-amyloid and prion protein peptides is independent from their neurotoxic effect. AB - Synthetic peptides corresponding to residues 25-35 of beta-amyloid (beta 25-35) and 106-126 of prion protein (PrP 106-126) are amyloidogenic and cause neuronal death by apoptosis in vitro. We evaluated, in rat cortical neurons, the role of caspases activation in the peptides neurotoxicity by measuring of caspase-3 (CPP32) activity and applying a non-selective caspase inhibitor (z-VAD-fmk) or CPP32-specific inhibitor (Asp-Glu-Val-Asp-CHO (DEVD-CHO)). CPP32 was dose dependently activated by both peptides (2.5-50 microM). The caspase inhibitors completely abolished the CPP32 activation induced by the peptides. However, the neurotoxic effect was partially attenuated with z-VAD-fmk, while no antagonism was found with DEVD-CHO. Thus, although beta 25-35 and PrP 106-126 robustly activated CPP32, their neurotoxic effect was independent of this caspase activation. PMID- 11036198 TI - Post-translational reduction of cell surface expression of insulin receptors by cyclosporin A, FK506 and rapamycin in bovine adrenal chromaffin cells. AB - Long-term (>/=3 h) treatment of cultured bovine adrenal chromaffin cells with cyclosporin A (CsA) decreased cell surface (125)I-insulin binding by 62% in a concentration (IC(50)=18 microM)- and time (t(1/2)=16 h)-dependent manner, but did not change the K(d) value. FK506 (1 microM) or rapamycin (3 microM) treatment reduced (125)I-insulin binding. Western blot analysis showed that CsA treatment decreased insulin receptor (IR) beta-subunit level (t(1/2)=15 h) in membrane fraction, but did not alter total cellular levels of IR precursor and IR beta subunit. Internalization rate of cell surface IR measured by using brefeldin A, an inhibitor of vesicular exit from the trans-Golgi network, was comparable between non-treated and CsA-treated cells. Thus, CsA, FK506 and rapamycin inhibit peptidyl prolyl cis-trans isomerase activities of cyclophilin and FK506-binding protein, and down-regulate IR presumably by reducing cell surface externalization of IR. PMID- 11036199 TI - Modulation of the deactivation kinetics of a recombinant rat T-type Ca(2+) channel by prior inactivation. AB - Using patch clamp methods we have investigated the deactivation properties of the T-type Ca(2+) channel generated by expression of the rat alpha(1I) subunit in HEK293 cells. The amplitude of the repolarisation-induced tail current was strongly correlated (R=0. 998) with the current amplitude immediately prior to repolarisation. The rate of deactivation was voltage-dependent between -120 mV (tau(deact)=0.9+/-0.0 ms) and -60 mV (tau(deac)=3.3+/-0.5 ms). Interestingly, the rate of deactivation observed at -80 mV was clearly dependent on the level of inactivation induced immediately prior to repolarisation, with greater inactivation producing significantly slower deactivation. In contrast, the rate of deactivation appeared completely independent of the level of steady-state inactivation. Together these data indicate the presence of a tight relationship between the recent induction of inactivation of this T-type channel and its subsequent rate of deactivation. PMID- 11036200 TI - Amyloid-like inclusions in Huntington's disease. AB - Huntington's disease is a progressive, autosomal dominantly inherited, neurodegenerative disease that is characterized by involuntary movements (chorea), cognitive decline and psychiatric manifestations. This is one of a number of late-onset neurodegenerative disorders caused by expanded glutamine repeats, with a likely similar biochemical basis. Immunohistochemical studies on Huntington's disease tissue, using antibodies raised to the N-terminal region of huntingtin (adjacent to the repeat) and ubiquitin, have recently identified neuronal inclusions within densely stained neuronal nuclei, peri-nuclear and within dystrophic neuritic processes. However, the functional significance of inclusions is unknown. It has been suggested that the disease-causing mechanism in Huntington's disease (and the other polyglutamine disorders) is the ability of polyglutamine to undergo a conformational change that can lead to the formation of very stable anti-parallel beta-sheets; more specifically, amyloid structures. We examined, using Congo Red staining and both polarizing and confocal microscopy, post mortem human brain tissue from five Huntington's disease cases, two Alzheimer's disease cases and two normal controls. Brains from five transgenic mice (R6/2)(12) expressing exon 1 of the human huntingtin gene with expanded polyglutamine, and five littermate controls, were also examined by the same techniques. We have shown that some inclusions in Huntington's disease brain tissue possess an amyloid-like structure, suggesting parallels with other amyloid associated diseases such as Alzheimer's and prion diseases. PMID- 11036201 TI - Imipramine and phenelzine decrease glutamate overflow in the prefrontal cortex--a possible mechanism of neuroprotection in major depression? AB - Antidepressant drugs have been used for decades, but the neurobiological substrate of their efficacy is not completely understood. Although these drugs have well-established effects on monoamines, evidence is emerging that they may also affect other neurotransmitter systems. It has been shown that treatment with a wide range of antidepressants changes the binding characteristics of the N methyl-D-aspartate type of glutamate receptor. This change is delayed and occurs only in the cortex. The mechanism that triggers it is unknown. We hypothesized that N-methyl-D-aspartate receptor alterations may be due to changes in the dynamics of cortical excitatory amino acid release. Such changes are of particular interest in areas such as the prefrontal cortex, a region involved in stress responses and affected in major depression. We investigated the effects of two antidepressants with different modes of action, imipramine and phenelzine, on glutamate and aspartate outflow in rat prefrontal cortex and striatum. We showed that antidepressants significantly decreased stimulated glutamate outflow. The effect had a rapid onset, was sustained during chronic administration and was only seen in the prefrontal cortex. This change may initiate receptor alterations. Furthermore, if antidepressants can dampen states of hyperglutamatergic activity and the subsequent excitotoxicity, their chronic use may have a considerable neuroprotective potential in major depression. PMID- 11036202 TI - Cannabinoid 1 receptors are expressed in nociceptive primary sensory neurons. AB - Expression of cannabinoid 1 (CB1) and vanilloid 1 (VR1) receptor proteins was studied in adult, cultured rat dorsal root ganglion neurons. Immunostaining of CB1 receptors alone produced labelling in 57+/-2% of the cultured dorsal root ganglion neurons (n=3 cultures). The area of the labelled cells was between 200 and 800 microm(2) with an average of 527+/-68 microm(2). VR1 immunolabelling revealed immunopositivity in 42+/-6% of the total population of dorsal root ganglion neurons. Cells showing VR1-like immunopositivity had an area between 200 and 600 microm(2). The mean area of the VR1-like immunopositive neurons was 376+/ 61 microm(2). Double immunostaining with antisera raised against the CB1 and VR1 receptor proteins, showed a high degree of co-expression between CB1 and VR1 receptors. An average of 82+/-3% of the CB1-like immunopositive cells also showed VR1-like immunoreactivity (n=3 cultures) while 98+/-2% of the VR1-like immunolabelled neurons showed CB1 receptor-like immunostaining (n=3 cultures). Our data suggests that nociceptive primary sensory neurons co-express CB1 and VR1 receptors to a very high degree. We propose that this may provide an anatomical basis for a powerful combination of VR1 mediated excitation and CB1-mediated inhibition of nociceptive responses at central and peripheral terminals of nociceptive primary afferents. PMID- 11036203 TI - Dopamine D5 receptors of rat and human brain. AB - In contrast to dopamine D1 receptors, the anatomical distribution of D5 receptors in the CNS is poorly described. Therefore, we have studied the localization of dopamine D5 receptors in the brain of rat and human using our newly prepared subtype-specific antibody. Western blot analysis of brain tissues and membranes of cDNA transfected cells, and immunoprecipitation of brain dopamine receptors suggest that this antibody is highly selective for native dopamine D5 receptors. The D5 antibody labeled dopaminergic neurons of mesencephalon, and cortical and subcortical structures. In neostriatum, the D5 receptors were localized in the medium spiny neurons and large cholinergic interneurons. The D5 labeling in caudate nucleus was predominantly in spines of the projection neurons that were frequently making asymmetric synapses. Occasionally, the D5 receptors were also found at the symmetric synapses. Within the cerebral cortex and hippocampus, D5 antibody labeling was prominent in the pyramidal cells and their dendrites. Dopamine D5 receptors were also prominent in the cerebellum, where dopamine innervation is known to be very modest. Differences in the localization of D5 receptors between both species were generally indistinguishable except in hippocampus. In rat, the hippocampal D5 receptor was concentrated in the cell body, whereas in human it was also associated with dendrites. These results show that D5 receptors are localized in the substantia nigra-pars compacta, hypothalamus, striatum, cerebral cortex, nucleus accumbens and olfactory tubercle. Furthermore, the presence of D5 receptors in the areas of dopamine pathways suggests that this receptor may participate actively in dopaminergic neurotransmission. PMID- 11036204 TI - Immunohistochemical localization of glial cell line-derived neurotrophic factor in the human central nervous system. AB - Glial cell line-derived neurotrophic factor, initially purified from the rat glial cell line B49, has the ability to promote the survival and differentiation of various types of neurons in the central and peripheral nervous systems. In the present study, to evaluate the physiological role of glial cell line-derived neurotrophic factor in the central nervous system, we investigated the cellular and regional distribution of glial cell line-derived neurotrophic factor immunoreactivity in autopsied control human brains and spinal cords using a polyclonal glial cell line-derived neurotrophic factor-specific antibody. On western blot analysis, the antibody reacted with recombinant human glial cell line-derived neurotrophic factor, and recognized a single band at a molecular weight of approximately 34,000 in human brain homogenates. Glial cell line derived neurotrophic factor immunoreactivity was observed mainly in the neuronal somata, dendrites and axons. In the telencephalon, diencephalon and brainstem, the cell bodies and proximal processes of several neuronal subtypes were immunostained with punctate dots. Furthermore, immunopositive nerve fibers were also observed, and numerous axons were intensely immunolabeled in the internal segment of the globus pallidus and the pars reticulata of the substantia nigra. In the cerebellum, the most conspicuous immunostaining was found in the Purkinje cells, in which the somata and dendrites were strongly immunolabeled. Intense immunoreactivity was also detected in the posterior horn of the spinal cord. In addition to the neuronal elements, immunopositive glial cell bodies and processes were observed in various regions. Our results suggest that glial cell line derived neurotrophic factor is widely localized, but can be found selectively in certain neuronal subpopulations of the human central nervous system. Glial cell line-derived neurotrophic factor may regulate the maintenance of neuronal functions under normal circumstances. PMID- 11036205 TI - Induction of vascular endothelial growth factor receptors and phosphatidylinositol 3'-kinase/Akt signaling by global cerebral ischemia in the rat. AB - Vascular endothelial growth factor is an angiogenic peptide that binds to tyrosine kinase receptors on target cells to activate signal transduction pathways involving phosphatidylinositol 3'-kinase and the serine-threonine protein kinase, Akt. To determine whether this signaling pathway is activated in cerebral ischemia, we examined the expression of vascular endothelial growth factor receptors 1 and 2, and phosphatidylinositol 3'-kinase-activated phospho Akt, in the cerebral cortex and hippocampus following transient global cerebral ischemia in the rat. Western blot analysis and immunocytochemistry demonstrated induction of vascular endothelial growth factor receptor 1 and 2 expression, and of anti-phosphatidylinositol 3'-kinase-immunoprecipitated phospho-Akt, in vulnerable regions of the cortex and hippocampus after 15 min of global ischemia and 4-72 h of reperfusion. These findings demonstrate that vascular endothelial growth factor receptors and receptor-coupled signal transduction pathways are induced in ischemic brain in vivo, and could therefore participate in endogenous neuroprotective responses to ischemia. PMID- 11036206 TI - Involvement of cerebral cortical structures in the classical conditioning of eyelid responses in rabbits. AB - The classical conditioning of the eyelid motor system in alert behaving rabbits has been used to study the expression of Fos in the hippocampus, and in the occipital, parietal, piriform and temporal cortices. Animals were classically conditioned with both delay and trace conditioning paradigms. As conditioned stimulus, both short and long (20 and 100 ms) tones (600 Hz, 90 dB) or short, weak (20 ms, 1kg/cm(2)) air puffs were used. The unconditioned stimulus was always a long, strong (100 ms, 3 kg/cm(2)) air puff that started 250-270 ms after the onset of the conditioned stimulus. The expression of Fos was significantly increased after both delayed and trace conditioning in the hippocampus, and in the parietal and piriform cortices contralateral to the unconditioned stimulus presentation side, compared with equivalent ipsilateral structures in conditioned animals, or with Fos production in the same contralateral structures in pseudo conditioned and control animals. Fos expression in some cortical sites was specific to tone versus air puff stimuli when used as conditioned stimulus. Thus, Fos expression was significantly increased in the contralateral temporal lobe when tones were used as conditioned stimulus, for both delayed and trace conditioning paradigms, but not when animals were conditioned to short, weak air puffs. The present results indicate a specific Fos activation in several cerebral cortical structures during associative eyelid conditioning. PMID- 11036207 TI - Neuropeptide Y Y5 receptor protein in the cortical/limbic system and brainstem of the rat: expression on gamma-aminobutyric acid and corticotropin-releasing hormone neurons. AB - Neuropeptide Y displays diverse modes of action in the CNS including the modulation of cortical/limbic function. Some of these physiological actions have been at least partially attributed to actions of neuropeptide Y on the Y5 receptor subtype. We utilized an antibody raised against the Y5 receptor to characterize the distribution of this receptor subtype in the rat cortical/limbic system and brainstem. Y5-like immunoreactivity was located primarily in neuronal cell bodies and proximal dendritic processes throughout the brain. In the cortex, Y5 immunoreactivity was limited to a subpopulation of small gamma-aminobutyric acid interneurons (approximately 15 microm diameter) scattered throughout all cortical levels. Double label immunofluorescence was also used to demonstrate that all of the Y5 immunoreactive neurons in the cortex displayed intense corticotropin releasing hormone immunoreactivity. The most intense Y5 immunoreactive staining in the hippocampus was located in the pyramidal cell layer of the small CA2 subregion and the fasciola cinerea, with lower levels of staining in the hilar region of the dentate gyrus and CA3 subregion of the pyramidal cell layer. Nearly all of the Y5 immunoreactive neurons in the hilar region of the hippocampus displayed gamma-aminobutyric-acid immunoreactivity. In the brainstem, Y5 immunoreactivity was most intense in the Edinger-Westphal nucleus, locus coeruleus and the mesencephalic trigeminal nucleus. The present study provides neuroanatomical evidence for the possible sites of action of the neuropeptide Y/Y5 receptor system in the control of cortical/limbic function. The presence of Y5 immunoreactivity on cell bodies and proximal dendritic processes in specific regions of the hippocampus suggests that this receptor functions to modulate postsynaptic activity. These data also suggest that the neuropeptide Y/Y5 system may play a role in the modulation of a specific population of GABAergic neurons in the cortex, namely those that contain corticotropin releasing hormone. The location of the Y5 receptor immunoreactivity fits with the known physiological actions of neuropeptide Y and this receptor. PMID- 11036208 TI - Dopamine and noradrenaline efflux in the prefrontal cortex in the light and dark period: effects of novelty and handling and comparison to the nucleus accumbens. AB - We used on-line microdialysis measurements of dopamine and noradrenaline extracellular concentrations in the medial prefrontal cortex of awake, freely moving rats during the dark and the light period of the day to study whether (i) basal efflux would be higher in the active, dark period than in the inactive, light period; (ii) the activation induced by environmental stimuli would be dependent on these conditions. When determined one day after cannula placement, noradrenaline and dopamine levels were higher during the dark. Maximal relative increases induced by novelty and handling were 150% and 175-200%, respectively, and were very similar in the light and the dark, but the net increases were higher in the dark. Separate groups were tested one week after cannula placement to ensure recovery of possibly disturbed circadian rhythms. While basal levels in the dark were now approximately twice those in the light, the maximal relative and net increases after both novelty and handling were very similar. Basal levels of dopamine in the nucleus accumbens (one day after cannula placement) were not different in the light or dark, but were increased by novelty and handling to about 130% only in the light period, not in the dark. Thus, in the prefrontal cortex, dopamine strongly resembles noradrenaline, in that basal efflux was state dependent, whereas activation by stimuli was not. In the nucleus accumbens, basal dopamine efflux was not state dependent, but activation by stimuli was. These results suggest that there are differential effects of circadian phase on basal activity and responsiveness of the mesolimbic vs the mesocortical dopamine system. PMID- 11036209 TI - Behavioral, neurochemical and endocrinological characterization of the early social isolation syndrome. AB - Rearing rats in isolation has been shown to be a relevant paradigm for studying early life stress and understanding the genesis of depression and related affective disorders. Recent studies from our laboratory point to the relevance of studying the social isolation syndrome as a function of home caging conditions. Accordingly, the present series of experiments assessed the contribution of each condition to the expression of the prepulse inhibition of the acoustic startle, food hoarding and spontaneous locomotor activity. In addition, ex vivo neurochemical changes in the brains of isolated and grouped rats reared either in sawdust-lined or in grid-floor cages were determined by measuring dopamine and serotonin as well as their major metabolites in a "psychosis circuit" that includes mainly the hippocampus and selected hippocampal efferent pathways projecting towards the anterior cingulate and infralimbic cortices, nucleus accumbens, dorsolateral caudate nucleus, amygdala and entorhinal cortex. The results of the present study demonstrate that rearing rats in isolation (i) produces a syndrome of generalized locomotor hyperactivity; (ii) increases the startle response; (iii) impairs prepulse inhibition; (iv) tends to increase food hoarding behavior; (v) increases basal dopamine turnover in the amygdaloid complex; (vi) decreases basal dopamine turnover in the infralimbic part of the medial prefrontal cortex; and (vii) decreases basal turnover of serotonin in the nucleus accumbens. In the entorhinal cortex, dopamine neurotransmission seemed to be more sensitive to the caging conditions since a decreased basal turnover of dopamine was observed in grid-reared animals. Plasma corticosterone levels were also increased in grid-reared animals compared with rats reared in sawdust cages. Finally, isolates reared on grids showed a significant positive correlation between plasma corticosterone levels and dopamine in the left nucleus accumbens.Altogether, these results support the contention that there is a link between social isolation, attention deficit, spontaneous locomotor hyperactivity and reduced dopamine turnover in the medial prefrontal cortex. Furthermore, our data demonstrate that rearing rats in grid-floor cages represents a form of chronic mild stress associated with increased corticosterone levels, decreased basal turnover of entorhinal dopamine and increased dopamine activity in the left nucleus accumbens. Finally, a significant and selective decrease in the basal turnover of serotonin in the nucleus accumbens of isolated rats may be linked to the isolation-induced locomotor hyperactivity. PMID- 11036210 TI - Alteration of expression levels of neuronal nitric oxide synthase and haem oxygenase-2 messenger RNA in the hippocampi and cortices of young adult and aged cognitively unimpaired and impaired Long-Evans rats. AB - Neuronal nitric oxide synthase and haem oxygenase-2 are postulated to be important enzymes involved in neuronal transmission and modulation of free radical levels in neurons. Hippocampal and cortical neuronal nitric oxide synthase and haem oxygenase-2 expressions were compared in young adult (6 months) and aged (24-26 months) Long-Evans rats. Aged rats were assigned as either cognitively unimpaired or impaired based on their performances in the Morris water maze behavioural task. In situ hybridization revealed increased neuronal nitric oxide synthase messenger RNA levels in selected regions of the hippocampi and cortices of aged rats. Moreover, aged cognitively impaired animals showed significantly higher neuronal nitric oxide synthase messenger RNA expression than aged cognitively unimpaired animals in several brain regions. For haem oxygenase 2 mRNA expressions, both young and aged cognitively impaired rats showed increased expressions in hippocampi compared with aged cognitively unimpaired rats, while no difference was found in cortices between all three animal groups. The increase in neuronal nitric oxide synthase messenger RNA expression levels in the aged animals may be related to increased free radical production occurring in ageing. Alternatively, elevated neuronal nitric oxide synthase and haem oxygenase 2 messenger RNA expressions may represent compensatory responses to oxidative stress and age-related changes in neuronal functions. Regarding cognitive status, aged cognitively impaired rats showed significant spatial memory deficits relative to young and aged cognitively unimpaired rats. Our data suggest a correlation between age-related cognitive impairment and change in messenger RNA expressions for the neuronal nitric oxide synthase and haem oxygenase-2 systems in brain areas implicated in learning and memory processes. PMID- 11036211 TI - Brain-derived neurotrophic factor delays hippocampal kindling in the rat. AB - Epileptic seizures increase the expression of brain-derived neurotrophic factor in the hippocampus. Since this neurotrophin exerts modulatory effects on neuronal excitability in this structure, it may play an important role in hippocampal epileptogenesis. This question was addressed by studying the effects of chronic infusions of recombinant brain-derived neurotrophic factor and brain-derived neurotrophic factor antisense in the hippocampus during the first seven days of hippocampal kindling. Infusion with brain-derived neurotrophic factor (6-24 microg/day) significantly delayed the progression of standard hippocampal kindling and strongly suppressed seizures induced by rapid hippocampal kindling. These suppressive effects were dose dependent, long lasting, not secondary to neuronal toxicity and specific to this neurotrophin, as nerve growth factor accelerated hippocampal kindling progression. They also appeared to be specific to the hippocampus, as infusion of brain-derived neurotrophic factor (48 microg/day) in the amygdala only resulted in a slight and transient delay of amygdala kindling. Conversely to the protective effects of exogenous brain derived neurotrophic factor, chronic hippocampal infusion of antisense oligodeoxynucleotides (12 nmol/day), resulting in reduced expression of endogenous brain-derived neurotrophic factor in the hippocampus, aggravated seizures during hippocampal kindling. Taken together, our results lead us to suggest that the seizure-induced increase in brain-derived neurotrophic factor expression in the hippocampus may constitute an endogenous regulatory mechanism able to restrain hippocampal epileptogenesis. PMID- 11036212 TI - Carbenoxolone depresses spontaneous epileptiform activity in the CA1 region of rat hippocampal slices. AB - An important contributor to the generation of epileptiform activity is the synchronization of burst firing in a group of neurons. The aim of this study was to investigate whether gap junctions are involved in this synchrony using an in vitro model of epileptiform activity. Hippocampal slices (400 microm) were prepared from female Sprague-Dawley rats (120-170 g). The perfusion of slices with a medium containing no added magnesium and 4-aminopyridine (50 microM) resulted in the generation of spontaneous bursts of population spikes of a fast frequency along with less frequent negative-going bursts. The frequency of the bursts produced was consistent over a 3h period. Carbenoxolone (100 microM), a gap junction blocker and mineralocorticoid agonist, perfused for 75 min, reduced the frequency of both types of spontaneous burst activity. Perfusion of spironolactone (1 microM), a mineralocorticosteroid antagonist, for 15 min prior to and during carbenoxolone perfusion did not alter the ability of carbenoxolone to depress the frequency of spontaneous activity. The incubation of hippocampal slices in carbenoxolone prior to recording increased the time taken for the spontaneous activity to start on change to the zero magnesium/4-aminopyridine medium and decreased the total number of spontaneous bursts over the first 60 min period. The ability of carbenoxolone to delay induction of epileptiform activity and reduce established epileptiform activity suggests that gap junctions contribute to the synchronization of neuronal firing in this model. PMID- 11036213 TI - GABAergic interneurons are the targets of cannabinoid actions in the human hippocampus. AB - Cannabinoids have been shown to disrupt memory processes in mammals including humans. Although the CB1 neuronal cannabinoid receptor was identified several years ago, neuronal network mechanisms mediating cannabinoid effects are still controversial in animals, and even more obscure in humans. In the present study, the localization of CB1 receptors was investigated at the cellular and subcellular levels in the human hippocampus, using control post mortem and epileptic lobectomy tissue. The latter tissue was also used for [3H]GABA release experiments, testing the predictions of the anatomical data. Detectable expression of CB1 was confined to interneurons, most of which were found to be cholecystokinin-containing basket cells. CB1-positive cell bodies showed immunostaining in their perinuclear cytoplasm, but not in their somadendritic plasmamembrane. CB1-immunoreactive axon terminals densely covered the entire hippocampus, forming symmetrical synapses characteristic of GABAergic boutons. Human temporal lobectomy samples were used in the release experiments, as they were similar to the controls regarding cellular and subcellular distribution of CB1 receptors. We found that the CB1 receptor agonist, WIN 55,212-2, strongly reduced [3H]GABA release, and this effect was fully prevented by the specific CB1 receptor antagonist SR 141716A. This unique expression pattern and the presynaptic modulation of GABA release suggests a conserved role for CB1 receptors in controlling inhibitory networks of the hippocampus that are responsible for the generation and maintenance of fast and slow oscillatory patterns. Therefore, a likely mechanism by which cannabinoids may impair memory and associational processes is an alteration of the fine-tuning of synchronized, rhythmic population events. PMID- 11036214 TI - Acquisition of an appetitive behavior reverses the effects of long-term treatment with lithium in rats. AB - Rats exposed to a long-term treatment with lithium chloride develop a deficit of avoidance accompanied by a reduction in the basal levels of extraneuronal dopamine and in dopamine accumulation in the nucleus accumbens shell after acute uptake inhibition. Such a condition is similar to that of an experimental model of depression induced by exposing rats to a chronic stress procedure. Rats exposed to chronic stress are also unable to acquire an appetitive behavior sustained by a highly palatable food. Thus, it was studied whether rats fed a diet containing lithium would develop an appetitive behavior induced by a pure hedonic stimulus. Rats on the lithium diet developed a clear-cut escape deficit condition accompanied by a decreased dopamine output in the nucleus accumbens shell; nevertheless, they learned the appetitive behavior within a similar period to controls. The development of the appetitive behavior coincided with the recovery of the capacity to avoid a noxious stimulus and with the return of the dopaminergic transmission in the nucleus accumbens shell to values similar to those of control rats. It may be concluded that the mechanism of action underlying the behavioral and neurochemical sequelae of a chronic stress is distinct from that of the analogous effects produced by lithium. PMID- 11036215 TI - Comparison of the fine structure of cortical and collicular terminals in the rat medial geniculate body. AB - Neurons throughout the rat medial geniculate body, including the dorsal and ventral divisions, display a variety of responses to auditory stimuli. To investigate possible structural determinants of this variability, measurements of axon terminal profile area and postsynaptic dendrite diameter were made on inferior colliculus and corticothalamic terminal profiles in the medial geniculate body identified by anterograde tracer labeling following injections into the inferior colliculus or cortex. Over 90% of the synapses observed were axodendritic, with few axosomatic synapses. Small (<0.5 microm(2)) and large (>1.0 microm(2)) collicular profiles were found throughout the medial geniculate, but were smaller on average in the dorsal division (0.49+/-0.49 microm(2)) than in the ventral division (0.70+/-0.64 microm(2)). Almost all corticothalamic profiles were small and ended on small-caliber dendrites (0.57+/-0.25 microm diameter) throughout the medial geniculate. A few very large (>2.0 microm(2)) corticothalamic profiles were found in the dorsal division and in the marginal zone of the medial geniculate. GABA immunostaining demonstrated the presence of GABAergic profiles arising from cells in the inferior colliculus. These profiles were compared with GABAergic profiles not labeled with anterograde tracer, which were presumed to be unlabeled inferior colliculus profiles or thalamic reticular nucleus profiles. The distributions of dendritic diameters postsynaptic to collicular, cortical and unlabeled GABAergic profiles were compared with dendritic diameters of intracellularly labeled medial geniculate neurons from rat brain slices. Our results demonstrate a corticothalamic projection to medial geniculate body that is similar to other sensory corticothalamic projections. However, the heterogeneous distributions of excitatory inferior collicular terminal sizes and postsynaptic dendritic diameters, along with the presence of a GABAergic inferior collicular projection to dendrites in the medial geniculate body, suggest a colliculogeniculate projection that is more complex than the ascending projections to other sensory thalamic nuclei. These findings may be useful in understanding some of the differences in the response characteristics of medial geniculate neurons in vivo. PMID- 11036216 TI - Characterization of synaptic transmission in the ventrolateral periaqueductal gray of rat brain slices. AB - Synaptic transmission evoked by focal stimulation in the ventrolateral periaqueductal gray was characterized using the whole-cell recording technique in rat brain slices. At resting membrane potential (-62+/-1 mV), focal stimulation (0.05-0.1 ms, 0.03 Hz) usually evoked a 6-cyano-7-nitroquinoxaline-2, 3-dione sensitive fast excitatory postsynaptic potential and a DL-2-amino-5 phosphonopentanoic acid-sensitive slow excitatory postsynaptic potential with a bicuculline-sensitive inhibitory postsynaptic potential in between. In the presence of kynurenic acid, bicuculline-sensitive inhibitory postsynaptic currents recorded in the voltage-clamp mode displayed a reversal potential of 68+/-3 mV, resembling GABA(A) receptor-mediated inhibitory postsynaptic currents. However, no GABA(B) receptor-mediated inhibitory postsynaptic current was evoked, even at stronger stimulating intensity. 6-Cyano-7-nitroquinoxaline-2,3-dione sensitive fast excitatory postsynaptic currents were isolated by DL-2-amino-5 phosphonopentanoic acid plus bicuculline and DL-2-amino-5-phosphonopentanoic acid sensitive slow fast excitatory postsynaptic currents by bicuculline plus 6-cyano 7-nitroquinoxaline-2,3-dione. Both types of excitatory postsynaptic current reversed at potentials near 0 mV. The I-V curve of slow fast excitatory postsynaptic currents or N-methyl-D-aspartate currents displayed a negative slope at potentials more negative than -30 mV in an Mg(2+)-sensitive manner. The control postsynaptic currents reversed at potentials between -50 and -35 mV, inclined to the reversal potential of GABA(A), but not glutamate, receptor channels. It is concluded that, in the ventrolateral periaqueductal gray, focal stimulation elicits both inhibitory and excitatory transmission, while the former is dominant. The inhibitory transmission is mediated by GABA(A) but not GABA(B) receptors. The excitatory transmission is mediated by glutamate acting on alpha amino-3-hydroxy-5-methylisoxazole-4-propionate/kainate as well as N-methyl-D aspartate receptors. PMID- 11036217 TI - Differential involvement of mu(1)-opioid receptors in endomorphin- and beta endorphin-induced G-protein activation in the mouse pons/medulla. AB - Several genetic mouse models of differential sensitivity to opioids have been used to investigate the mechanisms underlying individual variation in responses to opioids. The CXBK mice are inbred recombinant mice which have a lower level of mu(1)-opioid receptors than their parental strain. Endomorphin-1 and endomorphin 2 are endogenous opioid peptides that are highly selective for mu-opioid receptors, while beta-endorphin, which is also an endogenous opioid peptide, is non-selective for mu-, delta- and putative epsilon-opioid receptors. The present study was designed to investigate the effects of these endogenous opioid peptides on G-protein activation by monitoring guanosine-5'-o-(3-[35S]thio)triphosphate binding to pons/medulla membranes of CXBK mice and their parental strain C57BL/6 ByJ mice. Endomorphin-1 (0.1-10 microM), endomorphin-2 (0.1-10 microM) and beta endorphin (0.1-10 microM) increased guanosine-5'-o-(3-[35S]thio)triphosphate binding to the pons/medulla membranes from C57BL/6 ByJ and CXBK mice in a concentration-dependent manner. However, the increases of guanosine-5'-o-(3 [35S]thio)triphosphate binding induced by either endomorphin-1 or endomorphin-2 in CXBK mice were significantly much lower than those in C57BL/6ByJ mice. However, no significant difference was found in the increases of the guanosine-5' o-(3-[35S]thio)triphosphate binding induced by beta-endorphin in C57BL/6 ByJ and CXBK mice. Moreover, whereas the increase of guanosine-5'-o-(3 [35S]thio)triphosphate binding induced by 10 microM endomorphin-1 or endomorphin 2 were almost completely blocked by a mu-opioid receptor antagonist beta funaltrexamine (10 microM) in both strains, the increase of guanosine-5'-o-(3 [35S]thio)triphosphate binding induced by 10 microM beta-endorphin was attenuated to approximately 70% of stimulation by co-incubation with 10 microM beta funaltrexamine in both strains. The residual stimulation of [35S]guanosine-5'-o (3-thio)triphosphate binding by 10 microM beta-endorphin in the presence of 10 microM beta-funaltrexamine was further attenuated by the addition of putative epsilon-opioid receptor partial agonist beta-endorphin (1-27) (1 microM) in both strains. Like the endomorphins, the synthetic mu-opioid receptor agonist [D Ala(2),N-MePhe(4), Gly-ol(5)]enkephalin at 10 microM showed lower increases of guanosine-5'-o-(3-[35S]thio)triphosphate binding in CXBK mice than those in C57BL/6ByJ mice. However, there was no strain difference in the stimulation of guanosine-5'-o-(3-[35S]thio)triphosphate binding induced by 10 microM of the selective delta(1)-opioid receptor agonist [D-Pen(2,5)]enkephalin, delta(2) opioid receptor agonist [D-Ala(2)]deltorphin II or kappa-opioid receptor agonist U50,488H. The results indicate that the G-protein activation by endomorphin-1 and endomorphin-2 in the mouse pons/medulla is mediated by both mu(1)- and mu(2) opioid receptors. Moreover, beta-endorphin-induced G-protein activation in the mouse pons/medulla is, in part, mediated by mu(2)- and putative epsilon-, but not by mu(1)-opioid receptors. PMID- 11036218 TI - Expression of interleukin-6 receptor, leukemia inhibitory factor receptor and glycoprotein 130 in the murine cerebellum and neuropathological effect of leukemia inhibitory factor on cerebellar Purkinje cells. AB - Expression of glycoprotein 130 and the related receptors, including interleukin-6 receptor and leukemia inhibitory factor receptor, was examined in the murine cerebellum at the protein level. Western blot analysis revealed that interleukin 6 receptor, leukemia inhibitory factor receptor and glycoprotein 130 were expressed in the murine cerebellum. Immunoreactivities for interleukin-6 receptor, leukemia inhibitory factor receptor and glycoprotein 130 were strongly localized on the cell body of Purkinje cells, indicating that both interleukin-6 and leukemia inhibitory factor could act directly on Purkinje cells in murine adult mice. The expressions of interleukin-6 receptor, leukemia inhibitory factor receptor and glycoprotein 130 were observed on the cell membranes of Purkinje cells by immunoelectron microscopy. Immunoreactivity for the interleukin-6 receptor was also detected in the cytoplasm of Purkinje cells. Injection of a murine hemopoietic cell line, FDC-P1 cells, transfected with the complementary DNA encoding the leukemia inhibitory factor led to a reduction in calbindin positive dendrites of the Purkinje cells.The present results suggest that the leukemia inhibitory factor affects cerebellar functions through Purkinje cells. PMID- 11036219 TI - Changes in GABA-immunoreactivity during development of the rostral subdivision of the nucleus of the solitary tract. AB - GABA plays an important role in the processing of gustatory information in the rostral nucleus of the solitary tract. The following study used post-embedment immunohistochemistry in the rat brainstem to localize GABA at both the light and electron microscopic levels to characterize the developmental distribution of GABA and synaptogenesis of GABA-immunoreactive terminals in the rostral nucleus of the solitary tract. During the first postnatal week, GABA is present in the rostral nucleus of the solitary tract, but less of it is synaptic than any time later in development. Of the few synaptic terminals present at postnatal day 1, less than 20% are GABA-immunoreactive. This proportion more than doubles to reach adult levels by postnatal day 10. By weaning (postnatal day 20), GABA immunoreactive cells are found in nearly the same density as in the adult. Development continues after weaning and is characterized by a disproportionate loss of non-GABA-containing cells. Finally, one previously identified subtype of GABA-immunoreactive terminal matures very late during the postweaning phase of development. The study provides the first analysis of the development of GABA related circuitry in the rostral nucleus of the solitary tract using anatomical methods. These data provide the background with which to view the emerging physiology of developing taste neurons. PMID- 11036220 TI - Bi-directional changes in affective state elicited by manipulation of medullary pain-modulatory circuitry. AB - The rostral ventromedial medulla contains three physiologically defined classes of pain-modulating neuron that project to the spinal and trigeminal dorsal horns. OFF cells contribute to anti-nociceptive processes, ON cells contribute to pro nociceptive processes (i.e. hyperalgesia) and neutral cells tonically modulate spinal nociceptive responsiveness. In the setting of noxious peripheral input, the different cell classes in this region permit bi-directional modulation of pain perception (analgesia vs hyperalgesia). It is unclear, however, whether changes in the activity of these neurons are relevant to the behaving animal in the absence of a painful stimulus. Here, we pharmacologically manipulated neurons in the rostral ventromedial medulla and used the place-conditioning paradigm to assess changes in the affective state of the animal. Local microinjection of the alpha(1)-adrenoceptor agonist methoxamine (50.0 microg in 0.5 microl; to activate ON cells, primarily), combined with local microinjection of the kappa-opioid receptor agonist U69,593 (0.178 microg in 0.5 microl; to inhibit OFF cells), produced an increase in spinal nociceptive reactivity (i.e. hyperalgesia on the tail flick assay) and a negative affective state (as inferred from the production of conditioned place avoidance) in the conscious, freely moving rat. Additional microinjection experiments using various concentrations of methoxamine alone or U69, 593 alone revealed that the rostral ventromedial medulla is capable of eliciting a range of affective changes resulting in conditioned place avoidance, no place-conditioning effect or conditioned place preference (reflecting production of a positive affective state). Overall, however, there was no consistent relationship between place-conditioning effects and changes in spinal nociceptive reactivity. This is the first report of bi-directional changes in affective state (i.e. reward or aversion production) associated with pharmacological manipulation of a brain region traditionally associated with bi directional pain modulation. We conclude that, in addition to its well-described pain-modulating effects, the rostral ventromedial medulla is capable of modifying animal behavior in the absence of a painful stimulus by bi-directionally influencing the animal's affective state. PMID- 11036221 TI - Neutralizing antibodies against neurite growth inhibitor NI-35/250 do not promote regeneration of sensory axons in the adult rat spinal cord. AB - Neutralization of the myelin-associated neurite growth inhibitors NI-35 and NI 250 by IN-1 antibodies can promote axonal regeneration of several types of central nervous neurons. Here, we investigated in adult rats whether IN-1 can promote regeneration of ascending sensory axons across a peripheral nerve bridge back into the spinal cord. IN-1 was administered by hybridoma cells injected in the cerebral cortex or thoracic cord, its presence confirmed in tissue sections and cerebrospinal fluid, and its effectiveness demonstrated in co-cultures of oligodendrocytes and sensory neurons. With a two week infusion of control vehicle into the dorsal spinal cord 3 mm rostral to the nerve graft, only 3+/-2% of the anterogradely labeled sensory fibers present at the rostral end of the nerve graft had grown up to 0.5 mm, but not farther into the spinal cord. A similar limited extent of regeneration was seen with IN-1 or with infusion of Dantrolene, an inhibitor of NI-35/250 activity in vitro. With infusion of nerve growth factor rostral to the nerve graft, 40% of the fibers at the rostral end of the graft were found at 0.5 mm, 34% at 1 mm, 24% at 2 mm and 14% at 3 mm (the infusion site) into the spinal cord. Treatment with IN-l antibodies did not enhance the growth-promoting effects of nerve growth factor. We suggest that the neurite growth inhibitors NI-35 or NI-250 do not play a major inhibitory role in the regeneration of the ascending sensory fibers across a nerve bridge and back into the spinal cord of the adult rat. PMID- 11036222 TI - Prostaglandin E(2) enhances axonal transport and neuritogenesis in cultured mouse dorsal root ganglion neurons. AB - The effects of prostaglandin E(2) on axonal transport in cultured mouse dorsal root ganglion neurons were investigated by analysing the number of axonally transported particles under video-enhanced microscopy. Application of prostaglandin E(2) increased the number of particles transported in anterograde and retrograde directions. The EP(2) prostaglandin receptor agonist butaprost mimicked the effect of prostaglandin E(2), but the EP(1)/EP(3) prostaglandin receptor agonist 17-phenyl trinor prostaglandin E(2) and the EP(3) prostaglandin receptor agonist M&B 28767 had no effect. The membrane-permeable cyclic AMP analogue dibutyryl cyclic AMP and the adenylate cyclase activator forskolin mimicked the effect of prostaglandin E(2). The protein kinase A inhibitor H-89 reversibly reduced the number of particles in both anterograde and retrograde directions. The effects of prostaglandin E(2) and dibutyryl cyclic AMP were blocked by H-89. Taken together with previous biochemical studies showing that prostaglandin E(2) increases cyclic AMP levels, the present results suggest that prostaglandin E(2) enhances axonal transport via the EP(2) receptor and cyclic AMP-dependent protein kinase A pathway. We further investigated the role of prostaglandin E(2) in neurite growth. Prostaglandin E(2) increased both the number of cells exhibiting neurites and the neurite growth rate, operating by a similar mechanism to stimulation of axonal transport. Prostaglandin E(2) may modulate axonal transport to supply materials for morphogenesis as well as other functions in sensory neurons. PMID- 11036223 TI - Neuron-independent Ca(2+) signaling in glial cells of snail's brain. AB - To directly monitor the glial activity in the CNS of the pond snail, Lymnaea stagnalis, we optically measured the electrical responses in the cerebral ganglion and median lip nerve to electrical stimulation of the distal end of the median lip nerve. Using a voltage-sensitive dye, RH155, we detected a composite depolarizing response in the cerebral ganglion, which consisted of a fast transient depolarizing response corresponding to a compound action potential and a slow depolarizing response. The slow depolarizing response was observed more clearly in an isolated median lip nerve and also detected by extracellular recording. In the median lip nerve preparation, the slow depolarizing response was suppressed by an L-type Ca(2+) channel blocker, nifedipine, and was resistant to tetrodotoxin and Na(+)-free conditions. Together with the fact that a delay from the compound action potential to the slow depolarizing response was not constant, these results suggested that the slow depolarizing response was not a postsynaptic response. Because the signals of the action potentials appeared on the saturated slow depolarizing responses during repetitive stimulation, the slow depolarizing response was suggested to originate from glial cells. The contribution of the L-type Ca(2+) current to the slow depolarizing response was confirmed by optical recording in the presence of Ba(2+) and also supported by intracellular Ca(2+) measurement. Our results suggested that electrical stimulation directly triggers glial Ca(2+) entry through L-type Ca(2+) channels, providing evidence for the generation of glial depolarization independent of neuronal activity in invertebrates. PMID- 11036224 TI - Combination of molecular modeling and quantitative structure-activity relationship analysis in the study of antimycobacterial activity of pyridine derivatives. AB - A set of 4-benzylsulfanyl derivatives of pyridine-2-carbonitriles and pyridine-2 carbothioamides, previously tested for their antimycobacterial activity, were analysed by quantitative structure-activity relationship (QSAR) techniques, using some physicochemical and quantum-chemical parameters. The resulting QSAR revealed that the activity increases with electron withdrawing substituents in the benzyl moiety of studied compounds. HOMO orbitals can play an important role in the description of the mechanism of interactions at the molecular level. Additionally, the results of multiple linear regression indicate the differences between Mycobacterium tuberculosis and M. avium. The hydrophobicity of studied compounds is important for activity against M. avium. PMID- 11036225 TI - In vitro release of caffeine from concentrated W/O emulsions: effect of formulation parameters. AB - Concentrated water in oil emulsions have been obtained with four different emulsifiers to study the effect of formulation parameters on the in vitro release of caffeine. The in vitro release was studied on polysulfone membranes. Among the four emulsifiers, only one gave a statistically higher release of caffeine after 15 h (at a fixed percentage of dispersed phase). The concentration of the emulsifier does not have a significant effect on the release of caffeine. In contrast, diffusion of caffeine from concentrated W/O emulsions has been found to be highly dependent on the internal phase volume. The flux of caffeine increases with the percentage internal water phase. The droplet diameter decreases and the apparent viscosity increases with the percentage of the dispersed phase. And, the shape of the droplets goes from spherical to polyhedral as the percentage dispersed phase is increased. However, the flux could be correlated neither with the apparent viscosity nor with the droplet diameter at a fixed percentage of the dispersed phase. Results suggest that the shape factor may have an influence on the release of caffeine from concentrated emulsions. All the release profiles followed a zero-order kinetic. PMID- 11036226 TI - A comparison of the permeation enhancement potential of simple bile salt and mixed bile salt:fatty acid micellar systems using the CaCo-2 cell culture model. AB - The aim of this study was to compare the permeation enhancing potential and toxicity of simple bile salt and bile salt:fatty acid mixed micellar systems using the CaCo-2 cell culture model. The effects of micellar systems of sodium cholate, (NaC), and sodium taurocholate, (NaTC), on the permeability of the hydrophilic markers, mannitol (182) and polyethylene glycols (PEGS) 900 and 4000, were assessed. Simple micelle systems of the unconjugated bile salt, NaC, caused greater enhancement of the hydrophilic markers than the conjugated bile salt, NaTC. In the case of NaC systems the enhancement was coincident with excess membrane disruption and toxicity as indicated by altered TEERs, TEMs, MTT values, and, the lack of recovery following removal of the enhancer. In contrast, the NaTC systems were less toxic, and, in the simple micelle form the likely mechanism of enhancement of the hydrophilic markers is via a transient effect on the tight junctions. Formation of mixed micellar systems with linoleic acid (LA) accentuated the toxic effects of NaC. In comparison, NaTC:LA mixed micelles showed superior permeability enhancement versus simple micelles without increasing membrane toxicity. The mechanism of enhancement of NaTC:LA appears more complex and involves a possible combination effect on both the paracellular and transcellular routes. PMID- 11036227 TI - The selection of non-steroidal anti-inflammatory agents for dermal delivery. AB - An analysis has been conducted to show how the penetration of a selection of non steroidal anti-inflammatory agents (NSAIDs) through the skin may be predicted. The calculations are based on physicochemical parameters that can be predicted using commercially available software. Where available the predictions compare favourably with the literature values. The bio-effectiveness of the NSAID will be a function of both its penetration through the skin and its potency. The variation in potency has also been considered. Most NSAIDs are carboxylic acids, therefore the pK(a) will be an important determinant in ionisation and hence permeation. pH partition behaviour into the skin has been considered together with the relative impact of decreased permeation but increased solubility with degree of ionisation. PMID- 11036228 TI - Protective effect caused by the exopolymer excreted by Pseudoalteromonas antarctica NF(3) on liposomes against the action of octyl glucoside. AB - The capacity of the glycoprotein (GP) excreted by Pseudoalteromonas antarctica NF(3), to protect phosphatidylcholine (PC) liposomes against the action of octyl glucoside (OG) was studied in detail. Increasing amounts of GP assembled with liposomes resulted for the same interaction step in a linear increase in the effective surfactant to PC molar ratios (Re) and in a linear fall in the surfactant partitioning between bilayer and the aqueous phase (partition coefficients K). Thus, the higher the proportion of GP assembled with liposomes the lower the surfactant ability to alter the permeability of vesicles and the lower its affinity with these bilayer structures. In addition, increasing GP proportions resulted in a progressive increase of the free surfactant concentration (S(W)) needed to produce the same alterations in liposomes. The fact that S(W) was always lower than the surfactant critical micelle concentration indicates that the interaction was mainly ruled by the action of surfactant monomers, regardless of the amount of assembled GP. PMID- 11036229 TI - The novel combination of dynamic vapour sorption gravimetric analysis and near infra-red spectroscopy as a hyphenated technique. AB - The novel combination of an environmental controlled gas flow microbalance (Dynamic Vapour Sorption, Surface Measurement Systems, UK) with a NIR spectrometer (Foss NIR Systems) is described. The study follows the gravimetric changes and the spectroscopic changes in the amorphous and crystalline states of lactose at 298 K. NIR spectra and gravimetric water sorption were recorded simultaneously for the same sample. Differentiation of the amorphous and crystalline states of lactose was possible from the evaluation of peak intensity and shifts in the known fingerprint regions of the NIR spectra, i.e. 1350-1510 and 1825-1975 nm which correspond to water changes, and 2075-2160 nm which tends to illustrate changes in the organic/structural backbone character. Gravimetric analysis confirmed that the amorphous lactose crystallised, as weight changes can be linked to structural changes. The combined technique maintains the high performance of the DVS microbalance for gravimetric analysis but also provides a preset, regulated and controllable environment for studies using NIR spectroscopy probes, which was previously not possible. The results obtained agree with accepted data, and therefore provide validation for the hyphenation technique. The use of the combined DVS-NIR instrument has indicated two new pieces of information, firstly the amorphous form loses some water before the crystallisation is detectable. This indicates that water desorption may precede crystallisation, rather than the other way around, and secondly, the sample has completed crystallisation before water desorption has ended. PMID- 11036230 TI - The quantification of small degrees of disorder in lactose using solution calorimetry. AB - There is a realisation that small quantities of amorphous material can have a significant impact on the properties of crystalline solids. Consequently there is a growing interest in quantifying the amount of amorphous material that is present in "crystalline powders". Success has been reported when using isothermal microcalorimetry and vapour sorption techniques, however, the use of solution calorimetry has largely been ignored. In this study the enthalpies of solution of mixtures of amorphous and crystalline lactose are reported concentrating on the range 0-10% w/w amorphous content. It was found that there was a possible error due to water vapour penetration into the ampoule, resulting in crystallisation of the amorphous content, however this was overcome by double sealing the ampoules with wax. Subsequently there was a good correlation between the enthalpy of solution and the amorphous content, which was not adversely affected by stirring rate used during the experiment. Over the range from 0 to 10% amorphous content, quantification of the amorphous content of an unknown would be good to +/-0.5%. The effects of residual moisture retained within a sample were also investigated. Storage at 33% or 43% RH resulted in a much reduced wetting (exothermic) response compared with that seen for completely dry samples, which in turn led to a higher net enthalpy of solution. PMID- 11036231 TI - Preparation and in-vitro evaluation of poly[N-vinyl-2-pyrrolidone-polyethylene glycol diacrylate]-chitosan interpolymeric pH-responsive hydrogels for oral drug delivery. AB - Biocompatible and biodegradable pH-responsive hydrogels based on N-vinyl pyrrolidone (NVP), polyethylene glycol diacrylate (PAC) and chitosan were prepared for controlled drug delivery. These interpolymeric hydrogels were synthesized by a free radical polymerization technique using azobisisobutyronitrile (AIBN) as initiator and N,N'-methylenebisacrylamide (BIS) as crosslinker. These hydrogels were subjected to equilibrium swelling studies in enzyme-free simulated gastric and intestinal fluids (SGF and SIF). These swelling studies clearly indicated that these hydrogels were swollen more in SGF when compared to SIF. Theophylline and 5-fluorouracil (5-FU) were entrapped into these hydrogels and equilibrium-swelling studies were carried out for the drug entrapped gels in enzyme-free SGF and SIF. The in-vitro release profiles of the drugs were established in enzyme-free SGF. More than 50% of the entrapped drugs were released in the first 2 h at gastric pH and the rest of the drug release was slower. PMID- 11036232 TI - Dextran-methylprednisolone succinate as a prodrug of methylprednisolone: in vitro immunosuppressive effects on rat blood and spleen lymphocytes. AB - The in vitro immunosuppressive activity of a conjugate of methylprednisolone (MP) with dextran 70 kDa (DEX-MPS) was tested using the lymphocyte proliferation assay after stimulation of lymphocytes with concanavalin A (Con-A). Blood and spleen lymphocytes, isolated from drug-free male Sprague-Dawley rats, were used in the assay. First, the optimum concentration of Con-A for stimulation of lymphocytes was determined. The inhibition of the lymphocyte proliferation was then tested in the presence of 0.25,0.5, 1.0,2.5,5.0,10,20, and 50 nM concentrations (MP equivalent) of DEX-MPS or free MP. The maximum stimulation of lymphocytes with Con-A was observed at mitogen concentrations of 2.5 and 10 microg/ml for the spleen and blood lymphocytes, respectively. For free MP, sigmoidal relationships were observed between the effect (% inhibition of lymphocyte proliferation) and the logarithm of MP concentration. Additionally, the maximum inhibitory effect (I(max)) and MP concentration producing half of I(max) (IC(50)) were, respectively, 98% and 1.38 nM for the blood and 86% and 3.1 nM for the spleen lymphocytes. For MP conjugated to dextran, the response-log concentration curves were substantially shifted to the right with IC(50) values of 40 and 52 nM for the blood and spleen lymphocytes, respectively. It is concluded that compared with free MP, the steroid attached to dextran has minimal immunosuppressive activity. Therefore, to be effective in vivo, DEX-MPS should release MP in the body. PMID- 11036233 TI - The utilization of surface free-energy parameters for the selection of a suitable binder in fluidized bed granulation. AB - Surface free energy was determined for model substances pentoxyfilline, acyclovir, lactose and binding agents (that were used in the granulation process) hydroxypropilmethyl cellulose (HPMC) and polyvinylpyrrolidone (PVP) were determined by contact angle measurements. The methods of Wu, Good-van Oss and Della Volpe were used for solid-surface free-energy calculation. Spreading coefficients (S) were calculated and correlated with granulate properties. Granulates consisted of model drug and binding agent, and were produced in fluid bed granulator Glatt powder coater granulator GPCG1 by means of spraying the colloidal solution of binder on the model substance. Granules contained either 5% or 10% binder. Inverse granules, however, were also produced by spraying the model drug (i.e. pentoxyfilline and lactose) on the binding agent (HPMC, PVP). Particle size distribution, friability, true density, bulk density and tapped density of the granulates were determined. Although many different parameters influence the granule properties, it has been found that the interactions between the drug and the binder play a very important role. Spreading coefficients were found to be in good correlation with the friability of granulates. Positive spreading coefficient values of the binder over the model substance correlate well with the low friability of the granules containing lower amount of binder, i.e. 5%. In the group of the same binder, the spreading coefficient values decrease from pentoxyfilline over lactose to acyclovir. Friability results show that, for the system under consideration, PVP offers certain advantages over the grade of HPMC employed. The increase of the binder amount from 5 to 10% resulted in more friable granulates. Lower work of cohesion of the binder (PVP and HPMC) than the work of adhesion between binder and the model substances is considered responsible for the higher friability of the granules. The inverse granulation process, where the suspension of the model substance was sprayed over the solid binder particles, proved more efficient with HPMC than with PVP. According to the spreading coefficient results, the binder should spread over the drug. However, the kinetics of wetting appears to play an important role in the granulation process. According to these results, the conclusion was made that water wets HPMC much faster than PVP. PMID- 11036234 TI - Application of in vivo confocal microscopy to the objective evaluation of ocular irritation induced by surfactants. AB - An ocular irritation test using confocal laser scanning ophthalmoscopy has been developed in which corneal lesions subsequent to instillation of surfactants are specifically marked by fluorescein and assessed by digital image processing. The sum of the observed fluorescent corneal areas is taken into account as an endpoint of ocular irritation. Eight currently used nonionic, cationic and anionic surfactants were applied onto the cornea of rabbits and mice, four times per day during 3 days at various concentrations. Benzalkonium chloride, a cationic surfactant, at a concentration range of 0.01-0.5%, was tested in the same manner. The cornea was evaluated in vivo for ocular tolerance by confocal microscopy. In both rabbits and mice, the test revealed following irritation rankings: cationic>anionic>nonionic surfactants. Furthermore, in both animal models, the ocular damage increased with the concentration of benzalkonium. The test was sensitive enough to detect ocular microlesions at concentrations of surfactants as low as 0.01% for benzalkonium. These findings demonstrate the usefulness of confocal microscopy for the non-invasive, in situ evaluation of ocular tolerance. PMID- 11036235 TI - Effect of solvent on the preparation of surfactant-free poly(DL-lactide-co glycolide) nanoparticles and norfloxacin release characteristics. AB - The surfactant-free nanoparticles of poly(DL-lactide-co-glycolide) (PLGA) were prepared by dialysis method without surfactant and physicochemical properties such as particle size and drug contents were investigated against used initial solvent. The size of PLGA nanoparticles and drug contents were significantly changed by used initial solvent. The size of PLGA nanoparticles prepared from dimethylacetamide (DMAc), dimethylformamide (DMF), and dimethylsulfoxide (DMSO) as a initial used solvent was smaller than that of acetone. Selected initial solvent used to dissolve the copolymer significantly affects the size of nanoparticles and drug contents. It was shown that PLGA nanoparticles have spherical shapes from the results of scanning electron microscopy (SEM) and transmission electron microscopy (TEM) observations. It was thought that surfactant-free nanoparticles of PLGA entrapping norfloxacin (NFX) has nice drug loading capacity without free-drug on the surface of nanoparticles through the analysis of X-ray powder diffraction. From these results, it was showed the potential that the PLGA nanoparticles could be formed successively by dialysis method without surfactant. Release kinetics of NFX used as a model drug was governed by not only drug contents but also particle size parameter. The higher the drug contents and the larger the particle size resulted in slower the drug release. PMID- 11036236 TI - A new long acting ophthalmic formulation of carteolol containing alginic acid. AB - Alginic acid was evaluated as a potential vehicle in ophthalmic solutions for prolonging the therapeutic effect of carteolol. This anionic vehicle was expected to slow down drug elimination by the lacrimal flow, both by undergoing in-situ gel formation and by interacting with the mucus. In vitro studies indicated that carteolol is released slowly from alginic acid formulations, suggesting an ionic interaction. The adhesive behavior of alginic acid solution was better than that of another polymer, hydroxyethylcellulose (HEC). Intraocular pressure (IOP) measurements of rabbit eyes treated with a 1% carteolol formulation with or without alginic acid showed that this polymer significantly extended the duration of the pressure-reducing effect of carteolol to 8 h. The increased ocular bioavailability of 1% carteolol in the presence of alginic acid led to an equivalent concentration in the target tissue although administration was only once a day compared with twice a day for 1% carteolol alone. The overall results of this study indicate that the alginic-acid vehicle is an excellent drug carrier, well tolerated, and could be used for the development of a long-acting ophthalmic formulation of carteolol. PMID- 11036237 TI - Characterization of neuropeptide Y expression in the brain of a perciform fish, the sea bass (Dicentrarchus labrax). AB - The distribution of neuropeptide Y (NPY) gene expression was mapped in the brain of the sea bass (Dicentrarchus labrax) by in situ hybridization with 35S-UTP labeled cRNA probes. Gene expression was mainly detected within the forebrain, although NPY mRNA transcripts were also localized in the tectum and tegmentum mesencephali and posterior brain. New NPY-expressing nuclei were found in the dorsal and ventral telencephalon, preoptic area, tuberal hypothalamus, synencephalon, tegmentum mesencephali and posterior brain. The profuse NPY gene expression within the main neuroendocrine areas of the teleost fish further supports a physiological role in the control of the pituitary secretion. In addition, NPY gene was expressed within the primary visual, olfactory and gustatory circuits of teleost which, subsequently, project to hypothalamic feeding center in teleost fish. Our results extend the NPY-expressing areas known in teleost species. PMID- 11036238 TI - Localization of dopamine D1A and D1B receptor mRNAs in the forebrain and midbrain of the domestic chick. AB - The distribution and cellular localization of dopamine D1A and D1B receptor mRNAs in the forebrain and midbrain of the domestic chick were examined using in situ hybridization histochemistry with 35[S]-dATP labeled oligonucleotide probes, visualized with film and emulsion autoradiography. Labeling for D1A receptor mRNA was intense in the medial and lateral striatum, and moderately abundant in the pallial regions termed the archistriatum and the neostriatum, in the hypothalamic paraventricular nucleus region, and in the superficial gray layer of optic tectum of the midbrain. D1B receptor mRNA was abundant in the medial and lateral striatum, and in the pallial region termed the hyperstriatum ventrale, and moderately abundant in the intralaminar dorsal and posterior thalamus and in the superficial gray of the optic tectum. At the cellular level, about 75% of neurons in the medial striatum and 59% of neurons in the lateral striatum were labeled for D1A receptor mRNA, whereas about 39% of the neurons in the medial striatum and 21% in the lateral striatum were labeled for D1B receptor mRNA. Large striatal neurons were not labeled for D1A or D1B receptor mRNA. The data suggest that while both D1A and D1B receptors mediate dopaminergic responses in many neurons of the avian striatum, primarily D1A receptors mediate dopaminergic responses in the archistriatum and the neostriatum, while primarily D1B receptors mediate dopaminergic responses in the hyperstriatum ventrale and the thalamus. PMID- 11036239 TI - Regional distribution of PPARbeta in the cerebellum of the rat. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear receptors belonging to the superfamily of steroid hormone receptors. Different subtypes of PPARs (alpha, beta and gamma) have been described, PPARalpha and PPARgamma presenting a more tissue specific distribution than PPARbeta. Specific polyclonal antibodies directed against each subtype of PPARs were produced and characterized. The general expression of PPAR proteins was investigated in rat brain and cerebellar extracts by Western blotting. In order to localize the PPAR proteins and transcripts in the cerebellum, immunocytochemical and in situ hybridization assays were performed. Our Western blot analysis revealed a 52 kDa band with the anti-PPARbeta antibody in brain and cerebellar homogenates, but no band with the anti-PPARalpha, gamma1/gamma2 and gamma2. By immunocytochemistry, a high expression of PPARbeta appeared in the nucleus of Purkinje cells. The in situ hybridization assays showed that PPARbeta transcripts were localized in the cytoplasm of the Purkinje cells. No labeling was observed for the other PPAR isoforms in the cerebellum. Purkinje cells represent the only efferent way from the cerebellar cortex and modulate spinal cord activity. The regional distribution of PPARbeta in these cells suggests some fundamental role for this subtype in this pathway. PMID- 11036240 TI - Kappa opioid receptors are expressed by interneurons in the CA1 area of the rat hippocampus: a correlated light and electron microscopic immunocytochemical study. AB - A local GABA-system is known to have a mediatory function between several afferents and the principal cells of the hippocampus. This study examines the distribution and fine structure of kappa opioid receptor-immunoreactive elements in the CA1 subfield and reveals some new aspects concerning the structural basis of opioid-GABA interaction in the rat hippocampal formation. Kappa receptors were visualized immunocytochemically with a previously produced and characterized monoclonal antibody, the mAb KA8 (Maderspach, K., Nemeth, K., Simon, J., Benyhe, S., Szucs, M., Wollemann, M., 1991. A monoclonal antibody recognizing kappa-, but not mu- and delta-opioid receptors. J. Neurochem. 56, 1897-1904). The antibody selectively recognizes the kappa opioid receptor with preference to the kappa(2) subtype. Neuronal cell bodies, proximal dendrites and occasionally glial processes surrounding neuronal perikarya were labelled in the CA1 area. The immunopositive cells were present mainly in the stratum oriens, followed by the stratum pyramidale in a rostrocaudally increasing number. Their shape was fusiform, or multipolar. Occasionally kappa receptor-immunoreactive boutons surrounding weakly immunopositive somata were also observed. Electron microscopy of immunopositive neurons showed that the DAB labelling was intensive in the perinuclear cytoplasm. The widths and electron densities of the postsynaptic densities of some axosomatic synapses were remarkably increased. Similar increase of postsynaptic densities were observable at some axodendritic and axospinous synapses. On the basis of their location and fine structural properties the labelled cells are suggested to be GABAergic inhibitory interneurons, probably belonging to the somatostatinergic sub-population. The axons of these inhibitory interneurons are known to arborize in the stratum lacunosum-moleculare where the entorhinal afferents terminate. A modulatory effect of opioids on the entorhinal input, mediated by somatostatinergic interneurons is suggested PMID- 11036241 TI - Distribution of met-enkephalin immunoreactivity in the diencephalon and the brainstem of the dog. AB - The endogenous opioid system, in particular the enkephalins, has been implicated in a vast array of neurological functions. The dog could be a suitable model for the study of complex interactions between behavioral state and regulatory physiology in which the opioid system appeared to be implicated. Moreover, opiate derivatives are currently used in veterinary clinic and sometimes pharmacologically tested in the dog. However, there are no anatomical data regarding the organization of the opioid system in this species. The present work represents the first attempt to map the distribution of Met(5)-enkephalin-like immunoreactive (Met-enk-li) cell bodies and fibers in the diencephalon and the brainstem of the dog. In the diencephalon, labeled cells were present in all the mid-line and intralaminar thalamic nuclei; the lateral posterior, pulvinar and suprageniculate nuclei; the ventral nucleus of the lateral geniculate body and the medial geniculate body. Additionally, Met-enk-li cells were seen in every hypothalamic nucleus except in the supraoptic. Variable densities of labeled fibers were also seen in all these nuclei except in the medial geniculate body and in most areas of the lateral posterior and pulvinar nuclei. In the mesencephalon, positive cells were found in the periaqueductal gray, the Edinger Westphal and interpeduncular nuclei, delimited areas of the superior and inferior colliculi and the ventral tegmental area. In the rhombencephalon, labeled cells were seen in the majority of the nuclei in the latero-dorsal pontine tegmentum, the nuclei of the lateral lemniscus, the trapezoid, vestibular medial, vestibular inferior and cochlear nuclei, the prepositus hypoglossal, the nucleus of the solitary tract and the dorsal motor nucleus of the vagus, the infratrigeminal nucleus and the caudal part of the spinal trigeminal nucleus and in the rhombencephalic reticular formation. The distribution of fibers included additionally the substantia nigra, all the trigeminal nerve nuclei, the facial nucleus and a restricted portion of the inferior olive. These results are discussed with regard to previous reports on the distribution of Met-enk in other species. PMID- 11036242 TI - Magnetic resonance imaging of maxillary cancer--possibility of detecting bone destruction. AB - The purpose of this study was to evaluate the detectability of bone destruction of maxillary cancer with magnetic resonance imaging (MRI) using 14 cases of squamous cell carcinoma of the upper jaw. The detectability of bone destruction including the degree of spread to adjacent soft tissues was evaluated and compared to that of clinical examination, computed tomography (CT) and conventional X-ray films. MRI could show bone destruction of each bony part almost equally with CT, but differentiation among simple bone defects, bone expansion and bone destruction was difficult on MRI. The pattern of bone destruction of alveolus that could be detected on conventional X-ray examinations, could not be assessed on either CT or MRI. Soft tissue infiltration of the tumour was more clearly detected on MRI compared with CT and conventional X-ray films. PMID- 11036243 TI - Early stage carcinoma of oral tongue: prognostic factors for local control and survival. AB - The purpose of this study was to assess prognostic factors, treatment outcomes and patterns of relapse in patients with early stage (T1-2 N0) squamous cell carcinoma of oral tongue treated primarily by surgery. The medical records of all patients with early stage (T1-2 N0) oral tongue cancer, radically treated at King Faisal Specialist Hospital and Research Center between January 1980 and December 1997, were reviewed. Eighty-five patients were identified for analysis, 38 male and 47 female. With a median follow-up for surviving patients of 64 months, 5 year actuarial overall, disease-specific (DSS), and relapse-free survival (RFS) were 71, 75, and 63%, respectively. Univariate analysis for DSS showed survival advantage for patients with tumor thickness (TT) of < or =10 mm (P=0.0002) and distance from resection margin (DFRM) of > 5 mm (P=0.005). The effect of TT of < or =10 mm was maintained (P=0.001) on multivariate analysis. Higher RFS was observed with TT of < or =10 mm (P=0.0002), DFRM of > 5 mm (P=0.0002) and DFRM of >10 mm (P=0.007). On multivariate analysis higher RFS was also found for TT < or =10 mm (P=0.01) and DFRM >5 mm (P=0.01). Salvage of local tongue recurrence was higher than neck node failure, with 5-year DSS of 71 and 19%, respectively (P=0.007). Time interval for recurrence showed no significant impact on outcome. In T1-2 N0 oral tongue cancer, TT, and DFRM are significant prognostic factors for both local control and survival. Neck node recurrence is associated with poor prognosis and low salvage rate. PMID- 11036244 TI - Expression of E-cadherin, cellular differentiation and polarity in epithelial salivary neoplasms. AB - To investigate whether expression of E-cadherin correlates with polarised tissue organisation, grade or tumour type in salivary neoplasms, frozen sections from 30 salivary gland neoplasms were stained immunohistochemically for E-cadherin using the antibody HECD-1 and compared to the staining patterns in five samples of normal salivary gland. Lesions with areas of lack of staining were restained at two higher antibody concentrations. Normal salivary gland stained strongly around the periphery of acinar and ductal cells. Neoplasms mostly stained strongly regardless of neoplasm type. Reproducible loss of expression was found only in epithelial cells showing stromal or plasmacytoid (hyaline) differentiation in pleomorphic adenoma. Low- and high- grade mucoepidermoid carcinomas, adenocarcinoma NOS and carcinoma ex pleomorphic adenoma showed focal loss of expression but this was not related to tissue architecture, differentiation or invasiveness. We conclude that the relationship seen between E-cadherin expression and cell polarity/glandular organisation in breast and colon does not appear to exist for salivary gland neoplasms in which the diversity of architectural patterns precludes detection of any simple relationship. E-cadherin expression seems unlikely to be a useful marker for diagnosis or prognosis in salivary neoplasia in general. PMID- 11036245 TI - Primary intraoral leiomyosarcoma of the tongue: an immunohistochemical study and review of the literature. AB - Leiomyosarcoma is a relatively uncommon mesenchymal tumor that exhibits smooth muscle differentiation. We report a new case of leiomyosarcoma involving the tongue of a 67-year-old male. Histologically, the tumor was composed of variably oriented fascicles of spindle-shaped cells with 'cigar-shaped' nuclei and eosinophilic cytoplasm, containing occasional PAS-positive granules. Atypical mitotic figures and necrotic foci were frequently detected. Consistent desmin, alpha-smooth muscle-specific and sarcomeric actin, and vimentin immunoreactivity was demonstrated in the tumor cells, whereas cytokeratins, CD 30, CD 31, CD 34, CD 45, CD 68, EMA, GFAP, HMB 45 and S-100 protein were negative. The patient underwent wide surgical excision of the tumor and is alive and disease-free at a 5-year follow-up. This report emphasizes the difficulties in the differential diagnosis of these uncommon tumors in an intra-oral location. PMID- 11036246 TI - Detection of oral and oropharyngeal cancer by microsatellite analysis in mouth washes and lesion brushings. AB - Microsatellite allele losses are characteristic features of head and neck squamous cell carcinoma and can be used as molecular markers for malignancy. We have investigated the detection of microsatellite allele loss in mouth washes and lesions brushings from 19 patients with squamous cell carcinoma of the oral cavity and oropharynx as a means of tumour detection. In 84% of the analysed cases, allele loss previously identified in the tumour of these patients, was detected in these easily obtained specimens. No alterations were found in material derived from 10 healthy individuals. Success of detection was independent of tumour stage, suggesting that this approach may be useful for early diagnosis as well as for follow-up. PMID- 11036247 TI - Distinct patient groups in oral cancer: a prospective study of perceived health status following primary surgery. AB - Health-related quality of life (HRQOL) is an important consideration in the management of patients with cancer. Pre-treatment data can give an indication of the anticipated response following cancer treatment. However, in order to use HRQOL data meaningfully it is essential to have an appreciation of how different patient groups respond. The aim of the study was to identify distinct patient groups at baseline using the University of Washington head and neck cancer questionnaire (UW-QOL). It was also the intention to see how these groups differed in their: (1) clinical and demographic attributes; (2) European Organisation for Research and Treatment of Cancer (EORTC) C30 and SF 36 scores; and (3) longitudinal trends over 6 and 12 months. One-hundred and thirty consecutive patients with previously untreated oral and oro-pharyngeal cancer were recruited. All were treated by primary surgery. Attrition was evident in the study but care was taken to allow for this. There were two distinct groups of patients identified by the cumulative UW-QOL score at presentation. These groups discriminated throughout the EORTC C30 and SF 36, at baseline and longitudinally, with the exception of mental health. This study demonstrates that HRQOL items are interrelated. There are difficulties in using baseline scores for treatment selection, and scores need to be interpreted in the light of clinical and demographic factors. PMID- 11036248 TI - Thrombospondin-1 expression in oral squamous cell carcinomas: correlations with tumor vascularity, clinicopathological features and survival. AB - Thrombospondin-1 (TSP-1) is a 450 kd glycoprotein synthesized and incorporated into the extracellular matrix by numerous cell types and reported to suppress tumor growth and progression by its inhibition of angiogenesis. In order to clarify the biological role of TSP-1 and determine its clinicopathological significance in oral squamous cell carcinomas (SCCs), we identified TSP-1 protein expression in 54 oral SCCs by immunohistochemistry and correlated it with microvessel density (MVD), clinicopathological features and patient's survival. Thirty-two out of 54 carcinomas (59%) were identified as having a low level of TSP-1 expression (TSP-1-L), and 22/54 (41%) carcinomas identified as having a high level of TSP-1 expression (TSP-1-H). The MVD counts (mean+/-S.D.=9.0+/-4.9) in TSP-1-H tumors was significantly lower than that (mean+/-S.D.=12.7+/-4.4) in TSP-1-H tumors (P=0.0065). The level of TSP-1 expression was not correlated with T category and histologic grade, but inversely correlated with the pattern of tumor invasion (P=0.0136) and with lymph nodal status (P=0.0119). Furthermore, Kaplan-Meier analysis showed that the 5-year survival rate of TSP-1-H group was significantly higher than that of TSP-l-L group. Our results suggested that TSP-1 expression exerts an inhibitory effect on tumor vascularity, and that it has value in assessment of aggressiveness and prognosis of oral SCCs. PMID- 11036249 TI - Reduced expression of CD44 variant 9 is related to lymph node metastasis and poor survival in squamous cell carcinoma of tongue. AB - Expression of CD44v9 was immunohistochemically studied in 120 biopsy specimens from primary squamous cell carcinoma (SCC) of the tongue and correlated with clinicopathological findings of the SCCs. The tumors were classified into three groups according to immunostaining pattern of CD44v9; 53 cases with distinct positivity in all cancer cells except for those in the central part of nests (Group 1, non-reduced group), 42 cases with reduced expression in peripheral cells of nests (Group 2, reduced group), and 25 cases with complete disappearance of the expression in one or more nests (Group 3, negative group). Nineteen of 25 (76%) tumors in Group 3 and 14 of 42 (33%) in Group 2 exhibited lymph node metastasis, compared with only 8 of 53 (15%) in Group 1. The average survival time in Groups 1, 2 and 3 was 4496+/-204, 3866+/-379 and 2719+/-359 days, respectively and became shorter with the reduction of CD44v9 expression. These results suggest that the down-regulation of CD44v9 in SCC of the tongue may relate to the detachment of tumor cells from primary lesions, establishment of lymph node metastasis and consequently the death of patients. PMID- 11036250 TI - Exfoliative cytology of normal buccal mucosa to predict the relative risk of cancer in the upper aerodigestive tract using the MN-assay. AB - The high frequency of second or third primary tumors was first explained by Slaughter et al. with the concept of field cancerisation. Another theory postulates micrometastatic lesions as a reason for this phenomenon. The micronuclei (MN)-assay was evaluated to provide evidence for the concept of field cancerisation and to quantify the premalignant field change of normal mucosa in order to predict the individual cancer risk. MN-assay was carried out in 55 patients with squamous cell carcinoma of the head and neck, in 16 patients with a leucoplakia and in 99 healthy controls. A detailed questionnaire for population monitoring was completed. Buccal cytosmears of healthy mucosa of the study participants were examined for the MN count per 1000 cells. There was a direct correlation between tobacco abuse and increasing MN count as a sign of a cytogenetic damage of buccal mucosa cells. Alcohol did not influence the formation of MN. Both buccal sites were damaged in the same degree as proof of field cancerisation. The relative cancer risk (odds ratio) for smoking healthy controls with a definite MN frequency was estimated. Our study underscores the importance of the MN-assay as a biomarker to predict the relative cancer risk in the upper aerodigestive tract under suspicion of the individual susceptibility and the exposition to known carcinogenic agents such as tobacco and alcohol. The concept of field cancerisation was confirmed. PMID- 11036251 TI - Oral mucosal non-Hodgkin's lymphoma--a dangerous mimic. AB - Reports of T-cell lymphomas in the oral cavity are rare. Most have presented as a persisting ulcerated swelling. This paper reports two men, one of whom presented with a short history of increasing facial swelling and pain apparently related to a lower premolar tooth, and the other who had recurrent oral ulceration in several sites over a period of years. These types of cases are likely to present initially to general dental practitioners. PMID- 11036252 TI - Fat and free will. PMID- 11036253 TI - Resolving differences in GABAA receptor mutant mouse studies. PMID- 11036254 TI - Transforming growth factor-beta: death takes a holiday. PMID- 11036255 TI - Dendritic mRNA translation: deciphering the uncoded. PMID- 11036256 TI - Delivering the goods to synapses. PMID- 11036257 TI - Organization through experience. PMID- 11036259 TI - The neurosciences from A to Z PMID- 11036258 TI - Neural origins of 'I remember'. PMID- 11036260 TI - Self-administration behavior is maintained by the psychoactive ingredient of marijuana in squirrel monkeys. AB - Many attempts to obtain reliable self-administration behavior by laboratory animals with delta-9-tetrahydrocannabinol (THC), the psychoactive ingredient in marijuana, have been unsuccessful. Because self-administration behavior has been demonstrated in laboratory animals for almost all other psychoactive drugs abused by humans, as well as for nicotine, the psychoactive ingredient in tobacco, these studies would seem to indicate that marijuana has less potential for abuse. Here we show persistent intravenous self-administration behavior by monkeys for doses of THC lower than doses used in previous studies, but comparable to doses in marijuana smoke inhaled by humans. PMID- 11036261 TI - Parallel visuospatial and audiospatial working memory processes in the monkey dorsolateral prefrontal cortex. AB - The dorsolateral area of the prefrontal cortex (PFC) in primates is involved in visuospatial working memory, but the cellular basis of spatial working memory for auditory information is poorly understood. Here we examined dorsolateral PFC neurons using visual and auditory oculomotor delayed-response tasks. We found that the dorsolateral PFC contains two groups of neurons, each showing directional delay-period activity depending on the location of the visual or auditory cue, suggesting that parallel neuronal processes for visual and auditory spatial working memory occur in the dorsolateral PFC. PMID- 11036262 TI - Impaired recognition and experience of disgust following brain injury. AB - Huntington's disease can particularly affect people's recognition of disgust from facial expressions, and functional neuroimaging research has demonstrated that facial expressions of disgust consistently engage different brain areas (insula and putamen) than other facial expressions. However, it is not known whether these particular brain areas process only facial signals of disgust or disgust signals from multiple modalities. Here we describe evidence, from a patient with insula and putamen damage, for a neural system for recognizing social signals of disgust from multiple modalities. PMID- 11036263 TI - Two cis-acting elements in the 3' untranslated region of alpha-CaMKII regulate its dendritic targeting. AB - Dendritic localization of the alpha subunit of Ca2+/calmodulin-dependent protein kinase II (alphaCaMKII) mRNA in CNS neurons requires its 3' untranslated region (3'UTR). We investigated this targeting mechanism by identifying two cis-acting elements in the 3'UTR. One is a 30-nucleotide element that mediated dendritic translocation. A homologous sequence in the 3'UTR of neurogranin, transcripts of which also reside in dendrites, also funtioned in cis to promote its dendritic transport. Other putative elements in the alphaCaMKII mRNA inhibit its transport in a resting state. This inhibition was removed in depolarized neurons, and such activity-dependent derepression was a primary requirement for their dendritic targeting. PMID- 11036264 TI - Reduction of endogenous transforming growth factors beta prevents ontogenetic neuron death. AB - We show that following immunoneutralization of endogenous transforming growth factors beta (TGF-beta) in the chick embryo, ontogenetic neuron death of ciliary, dorsal root and spinal motor neurons was largely prevented, and neuron losses following limb bud ablation were greatly reduced. Likewise, preventing TGF-beta signaling by treatment with a TbetaR-II fusion protein during the period of ontogenetic cell death in the ciliary ganglion rescued all neurons that normally die. TUNEL staining revealed decreased numbers of apoptotic cells following antibody treatment. Exogenous TGF-beta rescued the TGF-beta-deprived phenotype. We conclude that TGF-beta is critical in regulating ontogenetic neuron death as well as cell death following neuronal target deprivation. PMID- 11036265 TI - Disruption of Eph/ephrin signaling affects migration and proliferation in the adult subventricular zone. AB - The subventricular zone (SVZ) of the lateral ventricles, the largest remaining germinal zone of the adult mammalian brain, contains an extensive network of neuroblasts migrating rostrally to the olfactory bulb. Little is known about the endogenous proliferation signals for SVZ neural stem cells or guidance cues along the migration pathway. Here we show that the receptor tyrosine kinases EphB1-3 and EphA4 and their transmembrane ligands, ephrins-B2/3, are expressed by cells of the SVZ. Electron microscopy revealed ephrin-B ligands associated with SVZ astrocytes, which function as stem cells in this germinal zone. A three-day infusion of the ectodomain of either EphB2 or ephrin-B2 into the lateral ventricle disrupted migration of neuroblasts and increased cell proliferation. These results suggest that Eph/ephrin signaling is involved in the migration of neuroblasts in the adult SVZ and in either direct or indirect regulation of cell proliferation. PMID- 11036266 TI - Postnatal synaptic potentiation: delivery of GluR4-containing AMPA receptors by spontaneous activity. AB - To examine how functional circuits are established in the brain, we studied excitatory transmission in early postnatal hippocampus. Spontaneous neural activity was sufficient to selectively deliver GluR4-containing AMPA receptors (AMPA-Rs) into synapses. This delivery allowed non-functional connections to transmit at resting potentials and required NMDA receptors (NMDA-Rs) but not CaMKII activation. Subsequently, these delivered receptors were exchanged with non-synaptic GluR2-containing AMPA-Rs in a manner requiring little neuronal activity. The enhanced transmission resulting from this delivery and subsequent exchange was maintained for at least several days and required an interaction between GluR2 and NSF. Thus, this sequence of subunit-specific trafficking events triggered by spontaneous activity in early postnatal development may be crucial for initial establishment of long-lasting functional circuitry. PMID- 11036267 TI - Dual MAP kinase pathways mediate opposing forms of long-term plasticity at CA3 CA1 synapses. AB - Although the function of the p42/p44 mitogen-activated protein (MAP) kinase pathway in long-term potentiation at hippocampal CA3-CA1 synapses has been well described, relatively little is known about the importance of the p38 MAP kinase pathway in synaptic plasticity. Here we show that the p38 MAP kinase pathway, a parallel signaling cascade activated by distinct upstream kinases, mediates the induction of metabotropic glutamate receptor-dependent long-term depression at CA3-CA1 synapses. Thus, two parallel MAP kinase pathways contribute to opposing forms of long-term plasticity at a central synapse. PMID- 11036268 TI - Adenoviral vector-mediated rescue of the OMP-null phenotype in vivo. AB - The use of gene deletion by homologous recombination to determine gene or protein function has wide application in vertebrate neurobiology. An ideal complement to gene deletion would be subsequent gene replacement to demonstrate re-acquisition of function. Here we used an adenoviral vector to replace the olfactory marker protein (OMP) gene in olfactory receptor neurons of adult OMP-null mice and demonstrated the subsequent re-acquisition of function. Our results show that short-term expression of OMP restores the kinetics of electrophysiological responses of OMP-null mice to those of the control phenotype. This adenoviral mediated rescue of the OMP-null phenotype is consistent with involvement of OMP in olfactory transduction. PMID- 11036269 TI - Ectopic synaptogenesis in the mammalian retina caused by rod photoreceptor specific mutations. AB - In addition to rod photoreceptor loss, many mutations in rod photoreceptor specific genes cause degeneration of other neuronal types. Identifying mechanisms of cell-cell interactions initiated by rod-specific mutations and affecting other retinal cells is important for understanding the pathogenesis and progression of retinal degeneration. Here we show in animals with rod and cone degeneration due to mutations in the genes encoding rhodopsin and cGMP phosphodiesterase beta subunit (PDE-beta) respectively, that rod bipolar cells received ectopic synapses from cones in the absence of rods. Thus, synaptic plasticity links certain rod specific mutations to retina-wide structural alterations that involve different types of neurons. PMID- 11036270 TI - Perception of Fourier and non-Fourier motion by larval zebrafish. AB - A moving grating elicits innate optomotor behavior in zebrafish larvae; they swim in the direction of perceived motion. We took advantage of this behavior, using computer-animated displays, to determine what attributes of motion are extracted by the fish visual system. As in humans, first-order (luminance-defined or Fourier) signals dominated motion perception in fish; edges or other features had little or no effect when presented with these signals. Humans can see complex movements that lack first-order cues, an ability that is usually ascribed to higher-level processing in the visual cortex. Here we show that second-order (non Fourier) motion displays induced optomotor behavior in zebrafish larvae, which do not have a cortex. We suggest that second-order motion is extracted early in the lower vertebrate visual pathway. PMID- 11036271 TI - BDNF upregulation during declarative memory formation in monkey inferior temporal cortex. AB - In primates, visual long-term memory of objects is presumably stored in the inferior temporal (IT) cortex. Because brain-derived neurotrophic factor (BDNF) is involved in activity-dependent neural reorganization, we tested the hypothesis that BDNF would be upregulated in IT cortex during formation of visual pair association memory. To eliminate genetic and cognitive variations between individual animals, we used split-brain monkeys for intra-animal comparison in PCR-based mRNA quantitation. The monkeys learned a pair-association (PA) task using one hemisphere and a control visual task using the other, to balance the amount of visual input. We found that BDNF was upregulated selectively in area 36 of IT cortex during PA learning, but not in areas involved in earlier stages of visual processing. In situ hybridization showed that BDNF-expressing cells were localized in a patchlike cluster. The results suggest that BDNF contributes to reorganization of neural circuits for visual long-term memory formation in the primate. PMID- 11036272 TI - Clustering of perirhinal neurons with similar properties following visual experience in adult monkeys. AB - The functional organization of early visual areas seems to be largely determined during development. However, the organization of areas important for learning and memory, such as perirhinal cortex, may be modifiable in adults. To test this hypothesis, we recorded from pairs of neurons in perirhinal cortex of macaques while they viewed multiple complex stimuli. For novel stimuli, neuronal response preferences for pairs of nearby neurons and far-apart neurons were uncorrelated. However, after one day of experience with the stimuli, response preferences of nearby neurons became more similar. We conclude that specific visual experience induces development of clusters of perirhinal neurons with similar stimulus preferences. PMID- 11036273 TI - Remembering episodes: a selective role for the hippocampus during retrieval. AB - Some memories are linked to a specific time and place, allowing one to re experience the original event, whereas others are accompanied only by a feeling of familiarity. To uncover the distinct neural bases for these two types of memory, we measured brain activity during memory retrieval using event-related functional magnetic resonance imaging. We show that activity in the hippocampus increased only when retrieval was accompanied by conscious recollection of the learning episode. Hippocampal activity did not increase for items recognized based on familiarity or for unrecognized items. These results indicate that the hippocampus selectively supports the retrieval of episodic memories. PMID- 11036274 TI - Neuronal activity in human primary visual cortex correlates with perception during binocular rivalry. AB - During binocular rivalry, two incompatible monocular images compete for perceptual dominance, with one pattern temporarily suppressed from conscious awareness. We measured fMRI signals in early visual cortex while subjects viewed rival dichoptic images of two different contrasts; the contrast difference served as a 'tag' for the neuronal representations of the two monocular images. Activity in primary visual cortex (V1) increased when subjects perceived the higher contrast pattern and decreased when subjects perceived the lower contrast pattern. These fluctuations in V1 activity during rivalry were about 55% as large as those evoked by alternately presenting the two monocular images without rivalry. The rivalry-related fluctuations in V1 activity were roughly equal to those observed in other visual areas (V2, V3, V3a and V4v). These results challenge the view that the neuronal mechanisms responsible for binocular rivalry occur primarily in later visual areas. PMID- 11036275 TI - Early colorectal cancer--concepts and clinical implications: introduction. PMID- 11036276 TI - Histopathology of early colorectal cancer. AB - The histopathologic spectrum of early colorectal cancer (ECC) is compared from the Western and Japanese perspective. In the West, most ECC presents as a malignant adenoma, whereas superficial ECC with no adenomatous component is well described in the Japanese literature. Furthermore, superficial ECC is thought to progress rapidly and disseminate early. The discrepant viewpoints are considered in the light of different approaches to histopathologic interpretation. Lesions reported by Japanese pathologists as superficial carcinomas limited to the mucosa are diagnosed as flat adenomas in the West. In one Japanese series only 4 of 218 (1.8%) flat or small sessile neoplasms removed by endoscopic mucosal resection were associated with submucosal invasion by carcinoma. Therefore superficial ECC with submucosal spread is uncommon. Most ECC in hereditary nonpolyposis colorectal cancer evolves through the adenoma-carcinoma pathway (including flat adenoma). However, adenoma removal in the context of population-based screening programs has not lowered the incidence of colorectal cancer; flat colorectal neoplasia may be important in clinical practice. PMID- 11036277 TI - Natural history of early colorectal cancer. AB - Since superficial tumors have been found, their peculiar pathologic features have evoked questions concerning their biologic behavior, their natural history. The aim of the present study was to elucidate the natural history of colorectal cancers (CRCs) including superficial cancers, using a retrospective radiologic method. Forty nine cancers that had had initial configurations of early cancer seen by previous radiography and that were examined pathologically were the subject of the present study. Growth speeds [doubling time (DT) calculation] and configurational changes at the various stages (invasion depth) were compared between polypoid growth (PG) and nonpolypoid growth (NPG). Growth speeds of mucosal cancer and submucosal cancer were also compared. The results showed that early CRC grows slowly (DT 31.2 months) when the cancer is limited to the mucosa. However, as tumors grow down to the submucosa, their growth speed accelerates (DT 25.8 months). The DT of these early cancers were longer than that of advanced cancers. The pathologic growth pattern (NPG or PG) of the CRCs did not affect the tumor growth speed. In respect to tumor configuration, when the tumor is limited to the submucosa the antecedent growth pattern may be easily deduced. It seems difficult, however, to know the initial growth patterns in advanced cancers because cancers with polyloid growth frequently change to a nonpolypoid growth pattern when in advanced stages. Among 32 advanced cancers, only 6 (19%) derived from IIc/IIc + IIa cancer. The most common (more than 70%) origin of advanced cancer seems to be IIa, Is, and Isp lesions. These results suggest that NPG cancers or superficial depressed cancers are not the main origins of advanced cancers, and that these cancer do not show extraordinarily rapid growth. PMID- 11036278 TI - Histoclinical analysis of early colorectal cancer. AB - To evaluate the clinicopathologic characteristics of early colorectal cancer (ECC), histopathologic and clinical features of 90 ECC patients who underwent surgical resection (not including the endoscopic polypectomized cases) and 1704 patients with advanced colorectal cancer were analyzed. Smaller size, better histologic differentiation, less lymph node (LN) metastasis, and better clinical outcome were observed in those with ECC than in patients with more advanced lesions. LN metastasis was found in 5 patients with ECC among the 56 patients who underwent bowel resection (8.9%). Tumors with LN metastasis were more frequently associated with deep submucosal invasion, absence of an adenomatous component within the tumor, sessile configuration, and lymphovascular invasion. Tumors showing deep submucosal layer invasion were associated with a more unfavorable histologic grade, lymphovascular invasion, LN metastasis, sessile morphology, and absence of an adenomatous component within the tumor. Recurrence was observed in two patients who underwent local excision for their submucosal cancer. One of the patients was salvaged after bowel resection, but one patient died of distant metastasis. From our data analysis and literature review, extensive submucosal invasion, lymphovascular invasion, sessile configuration, and tumors with no adenomatous component should be considered high risk factors for LN metastasis and recurrence after limited therapy. PMID- 11036279 TI - Radiographic diagnosis of early colorectal cancer, with special reference to the superficial type of invasive carcinoma. AB - The possibility of radiographically diagnosing carcinoma with submucosal involvement (invasive carcinoma) is discussed based on 119 invasive carcinomas of the large bowel that had been treated surgically and colonoscopically over a period of 9 years (1989-1997) at the Cancer Institute Hospital in Tokyo. Of these lesions, 38 were superficial-type invasive carcinoma, accounting for 31.9% (38/119) of all invasive colorectal carcinomas, including 36 lesions (94.7%, 36/38) of types IIa and IIa + IIc and 2 lesions (5.3%, 2/38) of type IIc + IIa. No pure type IIc was seen. The radiographic images obtained were correlated with macroscopic findings and analyzed in terms of visualization of the lesion's contour, central depression, converging folds, and basal indentation. A definitive diagnosis of superficial invasive carcinoma can be made radiographically if a lesion measures 10 mm and reveals moderate to severe basal indentation in a complete or nearly complete profile radiographic image. Attention should be paid to the presence of superficial-type advanced carcinomas measuring 10 mm or less, which is not infrequently experienced in Japan. Polypoid invasive carcinoma can be definitively diagnosed in the same way with much more certainty than can the superficial type because few polypoid advanced carcinomas are less than 20 mm. The size and the radiographic sign of the basal indentation are the most important indicators for the diagnosis of invasive carcinoma. PMID- 11036280 TI - Endoscopic management of polypoid early colon cancer. AB - Endoscopic management of polypoid early colonic cancer (malignant polyps and polypoid carcinomas) is no longer controversial. When the endoscopist is satisfied that excision is complete and histology is "favorable" (a resection margin of 2 mm and well or moderately well differentiated tumor), surgery is unnecessary. When histology show "unfavorable" characteristics (which a few histologists still take to include invasion into lymphatics), surgical or laparoscopic resection may be indicated, providing the patient is considered at suitable risk. Surgery kills some patients without finding residual cancer and cannot save others with metastases, so it should be recommended only with due clinical consideration. Sessile or broad-based polyps, especially those in the rectum, are more likely to be "high risk" and merit specialist management if local removal is to be attempted and to allow proper histologic assessment. Endoscopic approaches such as saline injection polypectomy, india-ink tattooing, and use of the argon beam coagulator are applicable in some cases. New approaches that still require trials include ultrasonographic probes, which occasionally clarify the degree of invasion, and prototype stapling devices to allow full thickness histologic specimens to be obtained. PMID- 11036281 TI - Surgical management of early colorectal cancer. AB - An early colorectal carcinoma is TNM stage T1NxMx. Most early carcinomas of the colon and rectum can be treated by adequate local excision, such as colonoscopic polypectomy and per-anal excision. If there are adverse risk factors, especially poorly differentiated carcinoma, lymphovascular invasion, or incomplete excision, a radical resection is indicated if there is no contraindication. In the case of a low rectal carcinoma, adjuvant chemoradiation should be considered. Recently a new classification has been developed: sm1 is invasion to the upper one-third of the submucosa, sm2 is invasion to the middle one-third, and sm3 is invasion to the lower one-third. Lesions of sm1 and sm2 have a low risk of local recurrence and lymph node metastasis; local excision is adequate. The sm3 lesions and sm2 flat and depressed types have a high risk of local recurrence and lymph node metastasis; further treatment is indicated. PMID- 11036282 TI - Role of laparoscopic surgery for treatment of early colorectal carcinoma. AB - The first laparoscopic colorectal surgery was performed in 1991. Several reports have showed some advantages for the laparoscopic technique compared to open procedures, but the lack of well designed trials has produced doubts about the real advantage of laparoscopy for colorectal cancer. To date, retrospective and prospective evidence suggests that laparoscopy is as safe as conventional colorectal surgery. Two recent prospective, randomized trials have showed that the short-term outcome is not compromised after laparoscopy for colorectal cancer. Furthermore, less pain, faster recovery of respiratory parameters, and better preservation of cell-mediated immune function have been associated with the laparoscopic technique. Laparoscopic surgery for early colorectal cancer may have a role in a well selected group of patients. PMID- 11036283 TI - Endorectal ultrasonography and treatment of early stage rectal cancer. AB - The purpose of this study was to evaluate the accuracy of preoperative staging by endorectal ultrasonography (EUS) and its contribution to treatment of early stage rectal cancer (ESRC). The results of EUS for 154 consecutive patients with ESRC (pTis to pT2) were compared prospectively with histologic findings, assessed according to the TNM classification. Results of treatment selection and long-term outcomes were analyzed retrospectively. There were 35 patients histologically staged as pTis, 8 as pT1-slight (invasion confined to the superficial one-third of the submucosa), 37 as pT1-massive (invasion extending to the deeper submucosa), and 74 as pT2. The equipment used was an echoendoscope GF-UM2 or GF UM3 (Olympus, Tokyo, Japan). Sensitivity/specificity/overall accuracy rates for detection of slight submucosal invasion, massive submucosal invasion, and muscularis propria invasion were 99%/74%/96%, 98%/88%/97%, and 97%/93%/96%, respectively. Incidences of lymph node metastasis in pTis, pTis to pT1-slight, pT1, pT1-massive, and pT2 cases were 0%, 0%, 18%, 22%, and 30%, respectively. Incidences of lymph node metastasis in ESRCs staged by EUS (u) as uTis, uT1 slight, uT1-massive, uT2, and uT3 by EUS were 0%, 0%, 26%, 36%, and 64%, respectively. Sensitivity, specificity, and overall accuracy rates for detection of positive nodes in overall ESRCs were 53%, 77%, and 72%, respectively. Of the 43 patients with pTis to pT1-slight tumors, 22 underwent endoscopic polypectomy or local excision, 20 radical surgery, and 1 radical surgery after endoscopic polypectomy due to vascular invasion. All these patients are alive and all but one (who refused radical surgery due to vascular invasion after local excision and developed liver and lung metastases) are disease-free. Of the 37 patients with pT1-massive tumors, 34 underwent radical surgery and 3 transcoccygeal segmental resection. All these patients are alive disease-free except for one who died of peritoneal carcinomatosis after radical surgery. All patients with pT2 tumors underwent radical surgery. The overall 5-year survival rates for pTis, pT1, and pT2 cases were 100%, 98%, and 97%, respectively. EUS is an accurate method for evaluating invasion depth in ESRC. Patients with uTis or uT1-slight tumors staged by EUS are at low risk of positive nodes and good candidates for endoscopic polypectomy or local excision. Those with uT1-massive or uT2 lesions should be treated with a radical operation because of the high incidence of positive nodes. PMID- 11036284 TI - Screening for early colorectal cancer. AB - There is now solid evidence from randomized trials suggesting that it is possible to reduce mortality from colorectal cancer by 15% to 25% by screening with fecal occult blood tests (FOBTs). The major benefit results from detection of early cancer in average-risk persons above 50 years of age who have a positive test followed by colonoscopy. However, it has to be demonstrated that the same acceptability can be reached in the general population as that obtained in trials. Many countries must establish a screening organization in a limited area to learn how satisfactory quality assurance can be obtained before a country-wide screening program is set up. So far, screening has not resulted in a reduced incidence of colorectal cancer in true population studies despite removal of two to three times as many possible precursors compared to controls. Cost effectiveness will probably be as good as that known from screening for breast cancer with mammography and better than that for cervical cancer. However, the calculations are based on the unhydrated Hemoccult-II test in randomized trials. More sensitive methods would be attractive, but none has yet been evaluated properly in average-risk persons. There is no general agreement how to screen high risk groups such as patients with previous colorectal adenomas and carcinomas, one or two first-degree relatives with colorectal neoplasia, or ulcerative colitis. Families with familial adenomatous polyposis or hereditary nonpolyposis colorectal cancer, however, are presented with firm guidelines. Genetic screening has been helpful in no more than these two small groups in the colorectal carcinoma universe. PMID- 11036285 TI - Early colorectal cancer with special reference to the superficial nonpolypoid type from a histopathologic point of view. AB - The incidence and histopathologic characteristics of nonpolypoid (superficial type) early colorectal carcinomas were studied and compared with those of the polypoid type. The superficial type was subclassified as elevated (type IIa), type IIa with central depression (type IIa + IIc), plain (type IIb), depressed (type IIc), and IIc with marginal elevation (type IIc + IIa). The superficial type comprised 22% and 27% of intramucosal and submucosal carcinomas, respectively. Pure type IIb was not found, and there were only three pure type IIc lesions. Type IIa + IIc and IIc + IIa (and IIc) showed a significantly higher rate of submucosal invasion among the small tumors (59% and 71% less than 20 mm, respectively) compared to the polypoid type; type IIa showed no significant difference. The incidence of lymph node metastasis among submucosal carcinomas showed no significant difference between the superficial type and the polypoid type. About 64% and 52% of type IIa and IIa + IIc tumors accompanied residual adenoma, suggesting that they originated from small, flat adenomas through the adenoma-carcinoma sequence, whereas type IIc + IIa (and IIc) did not have an adenomatous component, implying that they arose de novo or originated through an adenoma-carcinoma sequence at a smaller size than the type IIa and IIa + IIc lesions. Superficial-type early colorectal carcinomas are not rare, and they are not uniform in nature. Rapid growth and invasion to the submucosa is characteristic of superficial-type lesions with a central depression, and only superficial depressed (type IIc + IIa, IIc) lesions can arise de novo. Although they grow rapidly to invade the submucosa, it cannot be said that they show more aggressive behavior than the polypoid type. PMID- 11036286 TI - Colonoscopic diagnosis and management of nonpolypoid early colorectal cancer. AB - Nonpolypoid colorectal neoplasms are grossly classified into three groups: slightly elevated (small flat adenomas), laterally spreading, and depressed. Flat adenomas are not invasive until they are rather large, whereas depressed lesions can invade the submucosa even when they are extremely small. Nonpolypoid lesions are difficult to detect and are often overlooked. Keys to detect them are their slight color change, interruption of the capillary network pattern, slight deformation of the colonic wall, spontaneously bleeding spots, shape change of the lesion with insufflation and deflation of air, and interruption of the innominate grooves. Spraying of indigo carmine dye helps to clarify the lesions. A pit pattern analysis with a zoom colonoscope is useful for the diagnosis and staging of early colorectal cancer. Small flat adenomas are thought to be precursors of protruded polyps and lateral spreading tumors, whereas depressed lesions are thought to grow endophytically and become advanced cancers. Small depressed lesions are treated with an endoscopic mucosal resection (EMR) technique; but when they massively invade the submucosa, surgical resection is indicated. Laterally spreading tumors are not as invasive despite their large size and therefore are good indications for the EMR or piecemeal EMR method. Small flat adenomas need not be treated urgently, as almost none is invasive. Accurate diagnosis with dye-spraying and zoom colonoscopy is vital for deciding the treatment strategy. PMID- 11036287 TI - Colorectal carcinogenesis based on molecular biology of early colorectal cancer, with special reference to nonpolypoid (superficial) lesions. AB - The multistep genetic model of colorectal carcinogenesis is based on the concept of the adenoma-carcinoma sequence. The adenoma-carcinoma sequence theory has been generally accepted for polypoid early colorectal cancers (ECCs). On the other hand, an increasing number of nonpolypoid (superficial) ECC have been reported. Nonpolypoid (superficial) ECCs show distinct characteristics histologically and genetically, and some claim these lesions may develop by de novo type carcinogenesis. In fact, clinicopathologic studies have shown that most nonpolypoid (superficial) cancers have no adenomatous lesions in the surrounding area. Genetic analyses have also revealed that nonpolypoid (superficial) ECCs show a pattern of genetic alterations different from that of polypoid ECCs. The K ras mutation rate is lower in nonpolypoid (superficial) ECCs than in polypoid ECCs, but there is no significant difference in the p53 mutation rate between two types of tumor. During the development of ECCs, the K-ras gene seems to determine the macroscopic configuration: whether polypoid or nonpolypoid (superficial). These results suggest that nonpolypoid (superficial) ECCs originate from a pathway different from the conventional genetic pathway that follows the adenoma carcinoma sequence. However, this does not mean that this new pathway is following de novo type carcinogenesis, because there is a possibility that nonpolypoid (superficial) adenomas, or so-called flat adenomas, develop into nonpolypoid (superficial) ECCs following the adenoma-carcinoma sequence. At the present time, there is still not enough evidence to conclude whether nonpolypoid (superficial) ECC is derived from de novo carcinogenesis or the conventional adenoma-carcinoma sequence. Further analysis, especially concerning APC gene mutation in ECCs, is essential to elucidate the carcinogenesis of nonpolypoid (superficial) ECCs. PMID- 11036288 TI - Nonprotruding colorectal neoplasms: epidemiologic viewpoint. AB - Histologic sections from 781 nonprotruding colorectal neoplasias (adenomas and early carcinomas) recorded in Sweden and in Japan were reviewed by the same observer, using strict histologic criteria. Low grade dysplasia (LGD) was present in 82.8% (299/361) of the nonprotruding neoplasms recorded in Sweden but in only 42.6% (179/420) of those recorded in Japan. On the other hand, high grade dysplasia (HGD) was found in 42.4% (178/420) of the nonprotruding colorectal neoplasias in Japanese patients but in only 14.1% (51/361) of those in Swedish patients. Whereas 15.0% (63/420) of the nonprotruding neoplasms in Japan were intramucosal carcinomas (IMCs) or submucosal carcinomas (SMCs), only 3.0% (11/361) of those reviewed in Sweden were IMCs or SMCs. Thus in Japan the lesions were histologically more "advanced" (significantly more lesions had HGD) and more "aggressive" (significantly more lesions were IMC or SMC) than in Sweden. The cause(s) of that phenomenon remains elusive. The term nonpolypoid adenoma connotes a group of dysplastic lesions with different histologic phenotypes. Interestingly, similar phenotypes were recently found in overt colorectal carcinomas in both Sweden and Japan. Experimental and clinical findings indicate that nonprotruding colorectal adenomas are closely associated with colonic lymphoid nodules, suggesting that the lymphoid-associated colorectal mucosa might be the birthplace of many nonprotruding adenomas (and consequently of nonprotruding early colorectal carcinomas). PMID- 11036289 TI - Benefits of endoscopic vein harvesting. AB - The purpose of this study was to evaluate and compare the benefits of endoscopic saphenous vein harvesting (EVH) with the traditional incision technique (TIT) for coronary artery bypass grafting (CABG) in respect to the technical procedure and clinical outcome. In a prospective nonrandomized, case-matched study the greater saphenous vein was harvested for CABG in 22 patients using the endoscopic technique and in 18 patients with the traditional method. Comparisons were made for the operating time, length of incision and vein harvested, graft quality, postoperative complications, and pain assessment. Patient demographics were well matched. EVH required smaller incisions than did the TIT (10.5 +/- 6.6 vs. 31.2 +/- 7.8 cm, respectively; p < 0.0001). Harvest time and vein quality were comparable in the two groups. Total vein operating time was shorter following the endoscopic technique (60 +/- 24 vs. 100 +/- 35 minutes, respectively; p < 0.0001). EVH had fewer complications (NS), and postoperative pain was significantly less (p = 0.0034). The major advantages of endoscopic vein harvesting are a significant reduction of postoperative pain and strikingly better cosmetic results. Wound complications seem to be less frequent. PMID- 11036290 TI - Bedside percutaneous tracheostomy: prospective evaluation of a modification of the current technique in 100 patients. AB - Percutaneous dilatational tracheostomy (PDT) is being increasingly used. Concerns have been raised as to its safety, especially when it is done at the bedside. A prospective evaluation was conducted of 100 consecutive, unselected critically ill patients with PDT. The mean intensive care unit (ICU) stay before PDT was 12 days. One surgeon performed PDT alone (5 cases) or assisted residents (95 cases) in all operations; 84 were performed at the ICU bedside. Only the first six patients were taken to the operating room solely for tracheostomy. A modified technique was used: (1) the endotracheal tube was left in place during sequential dilations; (2) dilators were inserted in a 60-degree cephalad orientation to the skin and directed caudally after penetration of the anterior tracheal wall; (3) a digit was inserted through the tracheal opening to guide withdrawal of the endotracheal tube to the level of the vocal cords; and (4) size 8 tracheostomy cannulas were inserted over 28F dilators. The average time from skin incision to insertion of the tracheostomy tube was 12 minutes (< 10 minutes, 41 patients; 10 to 15 minutes, 37 patients; > 15 minutes, 22 patients). Sixty-five percent had unfavorable anatomic conditions due to spinal precautions or diffuse neck edema. Postoperative complications occurred in four patients; surgical emphysema after tracheal lacerations in three, cannula dislodgment in one. All complications were successfully managed without an operation by tube exchange (n = 3) or observation (n = 1); there was no procedure-related mortality. Forty patients were available for long-term follow-up (6-18 months after tracheostomy) by telephone; one had persistent hoarseness without respiratory difficulty. We concluded that bedside PDT is safe and easy to teach when performed with a technique that ensures correct instrumentation. PMID- 11036291 TI - Experimental study of the effect of nitric oxide inhibition on mesenteric blood flow and interleukin-10 levels with a lipopolysaccharide challenge. AB - The septic shock-induced decrease in mesenteric blood flow and release of proinflammatory cytokines are among the major pathophysiologic changes presumed to lead to multiple organ dysfunction syndrome (MODS). Increased nitric oxide (NO) levels are associated with both decreased mesenteric blood flow and positive modulation of proinflammatory cytokine release. In this study we aimed to determine the effect of the timing of the inhibition of nitric oxide synthase (NOS) on mesenteric blood flow and serum interleukin-10 (IL-10) concentrations during endotoxin shock. A nonspecific NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME), a specific NOS inhibitor aminoguanidine (AG), or placebo were injected 20 minutes before or 20 minutes after a lipopolysaccharide (LPS) or placebo challenge to Swiss-albino mice, as pretreatment or posttreatment, respectively. At 120 minutes after LPS or placebo injection the mesenteric blood flow was measured, and blood samples from the heart were obtained for IL-10 levels in both groups. Pretreatment and posttreatment with both NOS inhibitors prevented the LPS-induced decrease in mesenteric blood flow. Pretreatment was more effective for this purpose. Pretreatment accentuated the LPS-induced increase in serum IL-10 concentrations, whereas posttreatment had no significant effect. We conclude that the timing of NOS inhibition is important for attenuating some deleterious effects of endotoxin. PMID- 11036292 TI - Preoperative evaluation of pancreatic masses with positron emission tomography using 18F-fluorodeoxyglucose: diagnostic limitations. AB - Identification of pancreatic cancer in patients presenting with an enlarged pancreatic mass is a major diagnostic problem. Positron emission tomography (PET) using the radiolabeled glucose analogue 18F-fluorodeoxyglucose (FDG) has been suggested to provide excellent accuracy for noninvasive determination of suspicious pancreatic masses. We conducted a prospective study to verify these results. Forty-two patients admitted for pancreatic surgery underwent PET scanning. Image analysis was based on visual film evaluation and quantification of regional tracer uptake. PET imaging was visually analyzed by three observers blinded for the results of other diagnostic tests; they qualitatively graded the scans using a five-point scale (I = low to V = high) for the presence and intensity of focal FDG uptake. Diagnosis was proven by histology (n = 38) or follow-up (n = 4). Furthermore, the results of PET were compared with helical computed tomography (CT) and conventional ultrasonography (US), done during the routine diagnostic workup before pancreatic cancer surgery. Regarding only the results with scores of IV and V as positive for representing definite malignancy yielded a sensitivity of 71% and a specificity of 64% for film reading. Quantification of regional tracer uptake contributed no significant diagnostic advantage for differentiation between benign and malignant tumors. Helical CT revealed a sensitivity of 74% and a specificity of 45.5% and abdominal US 56% and 50%, respectively. We concluded that PET imaging provides only fair diagnostic accuracy (69%) for characterizing enlarged pancreatic masses. PET does not allow exclusion of malignant tumors. In doubtful cases, the method must be combined with other imaging modalities, such as helical CT. The results indicate that the number of invasive procedures is not significantly reduced by PET imaging. PMID- 11036293 TI - Appraisal of treatment strategy by staging laparoscopy for locally advanced gastric cancer. AB - More accurate preoperative staging is necessary to determine the treatment strategy for locally advanced gastric cancer. Thirty-two patients with T3 or T4 gastric cancer expected to undergo curative resection based on conventional examinations underwent staging laparoscopy. The disease stages determined were compared with those obtained by conventional methods. The discrepancy rate of disease staging was 16 of 32 (50.0%), with down-staging in 5 of 32 (15.6%) and up staging in 11 of 32 (34.4%). Of the 32 patients, 13 (40.6%) were found to have unsuspected peritoneal dissemination. The positive predictive value for peritoneal metastasis by staging laparoscopy was 100%, whereas the negative predictive value was 89% (17/19). The accuracy rate was 94%. After laparoscopy, 15 of the 32 (46.9%) were diagnosed as candidates for curative resection. Of these 15 patients who underwent surgery, 13 (86.7%) underwent curative resection (1 R0 and 12 R1); the remaining two underwent R2 resection because of peritoneal metastasis that was undetected by staging laparoscopy. Patients with tumors judged noncurable by laparoscopy (n = 11) received neoadjuvant chemotherapy. In 7 of the 11 cases, salvage surgery was done (one R0, three R1, three R2 resections). A second staging laparoscopy was performed in four cases to determine the indication for salvage surgery. Three of the four were judged to be curable and underwent curative resection. Staging laparoscopy is an effective tool for detecting unsuspected peritoneal metastasis, and it can increase the curative resection rate and decrease unnecessary laparotomy for advanced gastric cancer. Second-look laparoscopy enables accurate assessment of the chemotherapeutic response, which can help in decisions about salvage surgery. PMID- 11036294 TI - Improvement of operative mortality after curative resection for gastric cancer: population-based study. AB - It is not well known if the improvement in operative mortality after surgery for gastric cancer reported in hospital series can be extrapolated to the whole population. The aim of this study was to determine trends in operative mortality over a 20-year period in a nonselected community-based series of patients. A database of 648 patients with gastric cancer resected with curative intent between 1976 and 1995 in a region with a half-million population was divided into two periods: 1976-1983 and 1984-1995. Nonconditional logistic regression was performed to estimate the independent effects of the studied factors. Operative mortality was higher during the 1976-1983 period than during the 1984-1995 period (17.1% vs. 7.1%; p < 0.0001). When comparing the two study periods, operative mortality decreased dramatically from 26.2% to 10.0% in patients over age 70, from 31.8% to 7.9% after total gastrectomy, and from 30.7% to 6.3% after proximal esophagogastrectomy. Operative mortality after total gastrectomy was nearly the same as that after distal gastrectomy (7.9% vs 5.9%) during the second study period. During the first study period, operative mortality was independently associated with age at diagnosis, type of gastrectomy, and to a lesser degree stage at diagnosis; during the second study period, only age and stage at diagnosis were associated with the risk of operative mortality. This study indicates that in this well defined population operative mortality after curative resection for gastric cancer has decreased during the last 20 years. The results should encourage aggressive management of patients with gastric cancer, even in patients over 70 years of age. PMID- 11036295 TI - Postoperative biliary ascariasis: presentation and management--experience. AB - Subsequent to preoperative and perioperative indications the common bile duct was explored in 705 patients over a 12-year period, from January 1983 to December 1994. Consequent postoperative T-tube cholangiography revealed the presence of worms in 22 patients. Expulsion of the worms followed T-tube irrigation with 0.9% normal saline in 18 patients. Only one patient had to be reexplored to remove the ascaris. In two patients the worm was removed along with the T-tube, and in one patient the worm came out through the T-tube tract. PMID- 11036296 TI - Debridement of gunshot wounds: semantics and surgery. AB - Debridement is a well established modality for management of gunshot wounds. The word "debridement" is originally French. It was used for the first time during the eighteenth century in the surgical context and meant "wound incision." For French surgeons, it has retained to this day its original meaning. In medical English, though, the use of the term has been marred by persisting confusion about its definition. Two quite different surgical procedures still compete for the definition of debridement: wound incision and wound excision. These procedures are also at the center of a modern controversy about the management of gunshot wounds. The orthodox doctrine, inherited from military surgeons, consists of aggressive tissue excision around the bullet track. This radical policy is being challenged by advocates of a more conservative approach. Minimal tissue excision is sufficient and safe in many cases provided careful monitoring of the wound is instituted. Wound incision alone to relieve tension and allow drainage is possible in certain cases. The tug-of-war between excision and incision is outlined herein with reference to the semantic tribulations of the word "debridement" and the implications for patient care. PMID- 11036297 TI - [Antibacterial therapy of patients with inflammatory necrotic pancreatitis]. AB - The infectionized necrotic pancreatitis (NP) course, complicated by localized and diffusive peritonitis, abscess and retroperitoneal phlegmon in 86 patients, was analyzed. Severity of state of patients according to the APACHE II scale was estimated. Recommendations for application of empirical and purposeful antibacterial therapy was elaborated. Total mortality for infectionized NP with complicated course was 25.5%. PMID- 11036298 TI - [Contemporary approaches to conservative treatment of purulent necrotic complications of an acute pancreatitis]. AB - The experience of treatment of 860 patients with purulent-necrotic complications of an acute pancreatitis was summarized. The complex approach, the methods of an organism detoxication application, the operative procedure optimization permit to stop the purulent-necrotic process progress and to perform the operative intervention in the period of time, favourable for the patient. PMID- 11036299 TI - [Diagnosis of the pancreonecrosis using the gas chromatography method]. AB - In 62 patients with various forms of an acute pancreatitis (AP) the contents of methane, oxygen, nitrogen, carbon dioxide gas in the blood, using the gas chromatography method, was studied up. In 55 patients with pancreonecrosis the changes of concentration of methane, oxygen and nitrogen had correlated with the disease course severity, the data of ultrasound investigation and bolus computer tomography. Basing on the investigations performed in 62 patients with an AP, in 12--with other acute surgical diseases of abdominal cavity and in 20 healthy persons, it was established the possibility of determination of the methane level in the blood as a criterion of the various forms of pancreonecrosis diagnosis and the method of its diagnosis was proposed. PMID- 11036300 TI - [Prognostic value of oxygen metabolism indices in the diffuse purulent peritonitis complicated by sepsis]. AB - There were the results of examination of 42 patients with diffuse purulent peritonitis, complicated by sepsis, analyzed. High risk of the oxygen debt, occurring due to limitation of the oxygen delivery to tissues, was noted. Normalization of delivery consumption ratio of oxygen and elimination of the oxygen debt in the early course of the disease are needed for the disease severity reduction and the mortality lowering. PMID- 11036301 TI - [The application of hemosorption in the treatment of patients with obturative jaundice after cholecystectomy]. AB - In 47 patients there was applied the hemosorption (HS) in complex of treatment of obstructive jaundice (OJ) after cholecystectomy. The jaundice duration was 4-6 weeks at average. Choledocholithiasis, the papilla magna duodeni stenosis, choledochal cicatricial stricture were the most frequent causes of OJ. There were conducted 73 HS procedures. In 2 h the level of general bilirubin, cholesterol, urea, creatinine, biliary acids, phenols, middle molecular weight peptides, the alkaline phosphatase activity had lowered significantly. Positive changes of the indices were noted. One-two day term after the HS conduction is optimal one for the radical operation performance. PMID- 11036302 TI - [Benign tumors of the stomach and intestine as a cause of bleeding]. AB - There were examined 30 patients with benign tumor of stomach and intestine, complicated by hemorrhage. The operation conduction is indicated when benign tumor is revealed because of the profuse hemorrhage recurrency danger. When the bleeding gastric or intestinal polyp is present the conservative therapy or the conduction of endoscopic hemostasis and biopsy with subsequent performance of the planned operative intervention are effective. PMID- 11036303 TI - [Morphological changes of gastric mucosa in duodenal ulcer disease in elderly and senile patients]. AB - Morphological investigation of gastric mucosa was done in 85 patients of various age with duodenal ulcer disease using visual-analogue scale of the modified classification of the Sidney system (1996). In all the patients there was revealed chronic gastritis, possessing age morphological peculiarities. The frequency of infectionizing with Helicobacter pylori is reduced with the age increased. PMID- 11036304 TI - [Metachronous megacolon and colonic cancer in adults]. AB - The experience of treatment of 9 patients with metachronous megacolon and colonic cancer was analyzed. Clinical signs of the disease are diverse. Radical surgical intervention is singularis method of treatment of such patients. PMID- 11036305 TI - [Technical aspects of revascularizing transplantation of omentum majus on myocardium, upper extremity and brain]. AB - There was the technique elaborated for the revascularized transplantation of the omentum majus flap on myocardium, upper extremity and brain. There were performed 26 operations of nondirect revascularization of myocardium, 3--of microsurgical transplantation on upper extremity and 3--on the brain. PMID- 11036306 TI - [Laparoscopy in the diagnosis and treatment algorithm for an acute surgical diseases and trauma of abdomen]. AB - Laparoscopy was performed in 1977 patients in an acute surgical diseases and the abdominal cavity (AC) injuries, the curative one--in 895. In patients with clinical signs of the AC organs acute disease or trauma it is possible the four variants diagnostic and curative program application: radial methods of investigation and laparocentesis--for diagnosis, laparoscopy and laparotomy--for diagnosis and treatment. The laparoscopy application is indicated even if the "acute abdomen" clinical features presence are confirmed the radial diagnosis and laparocentesis result. If necessary the diagnostical laparoscopy is transformed into curative procedure, which is used with the miniinvasive access application, resulting in low operative risk. PMID- 11036307 TI - [Surgical treatment of the liver injury in combined closed thoracoabdominal trauma]. AB - The experience of surgical treatment of 513 injured persons with severe closed thoracoabdominal trauma (CTAT) in an acute period was summarized. In 142 (27.8%) patients the hepatic injury was revealed. The CTAT structure, the diagnosis and surgical tactics peculiarities in the liver injury, complications, the mortality outcome causes were analyzed. Lethality was 64.1%. PMID- 11036308 TI - [The experience of application of peritoneal dialysis in cardiosurgery of an early age children]. AB - The experience of the peritoneal dialysis (PD) successful application in 16 children of an early age was summarized. In all the patients the terminal state with an acute cardiac and renal insufficiency was noted after cardiosurgical operation conduction using artificial blood circulation. The improvement of hemodynamical indices, renal and pulmonary function as well, were promoted by an early application of PD. PMID- 11036309 TI - [The renal function disorders in children with peritonitis of appendicular genesis]. AB - In 36 children with the appendicular genesis peritonitis the changes of excretory, ionregulating and acidexcretorial renal functions were studied up, using clearance-method under the conditions of spontaneous nocturnal diuresis. The glomerular filtration rate, the endogenic creatinine concentrational and the sodium excretion indices were the most informative exponents. PMID- 11036310 TI - [Age-related changes of pulmonary surfactant in patients with acute peritonitis and their correction]. AB - The results of research of the superficial activity of pulmonary surfactant (PS) in 124 patients with peritonitis are given. In patients younger than 60 years old PS changes depended on the phase: initial decrease in the reactive phase, stabilization--in the toxic phase, severe decrease--in the terminal one. In patients older than 60 progressive decrease of PS activity was shown, which was a risk factor for the bronchopulmonary complications: pneumonia, athelectasis, acute respiratory failure. The complex of actions for the prophylaxis and correction of PS insufficiency in patients with peritonitis is proposed. The efficacy of use of the exogenic PS substance sucrim was shown. PMID- 11036311 TI - [Prophylaxis and treatment of postoperative complications in peritonitis of appendicular genesis]. AB - The results of treatment of postoperative complications in 227 children with destructive forms of appendicitis and appendicular peritonitis were analyzed. The expediency of the low-intensive laser radiation application in the treatment of various forms of peritonitis was noted. The frequency of the postoperative complications occurrence was 17.3% at average, constituting one half in comparison with the control groups. PMID- 11036312 TI - [Criteria of immunotherapy application and its efficacy control in postoperative purulent-inflammatory complications]. AB - In 136 patients with surgical diseases the dynamics of the immunity indices and cytological changes of the wound discharge after the operation were studied up. Twenty practically healthy persons had constituted the control group. The most informative immunogram indices, which may be used for the postoperative complications occurrence prognosis, were determined. The results of investigations of the cytological changes in the wound are reflecting the immunoinflammatory reaction adequacy and serve as criterion for the immunotherapy administration substantiation in its efficacy control. PMID- 11036313 TI - [Immune state of patients of various age with the thyroid gland tumor]. AB - The comparative analysis of immune state of the various age patients with benign and malignant thyroid gland (TG) tumors is presented. In the TG cancer the immunogenesis insufficiency is more pronounced. The data obtained trusts the perspectiveness of the immunocorrecting therapy application in patients with the TG cancer. PMID- 11036314 TI - [The change of biogenic amine blood level under the influence of the low intensity laser infrared radiation]. AB - The dynamics of the biogenic amines level in the blood of patients after performance of hemorrhoidectomy was dependent on severity of an organism stress reaction predicting the wound healing outcome. When the wound irradiation using the low-energy laser of infrared diapason is performed in patients with the lowered reactivity of organism the histamine and serotonin contents dynamics is the same as in the patients with normal reactivity. PMID- 11036315 TI - [Modern principles of conservative treatment of necrotic pancreatitis]. PMID- 11036316 TI - [Contemporary aspects of treatment of colonic cancer complicated by ileus. Part I. Classification, surgical strategies, results of treatment]. PMID- 11036317 TI - [The choice and substantiation of optimal strategies in the surgical treatment of thyroid cancer]. PMID- 11036318 TI - [Necrotic-inflammatory affection of foot in patients with diabetes mellitus. Terminology and classification]. PMID- 11036319 TI - [Congenital cystic duplication of stomach in a child simulating a tumor]. PMID- 11036320 TI - [Gastric perforation into the left pleural cavity after pleuropulmonectomy for pulmonary cancer]. PMID- 11036321 TI - [Jejunal leiomyoma as a cause of severe bleeding]. PMID- 11036322 TI - [The method of radical surgical treatment of acute relapsing paraproctitis]. PMID- 11036323 TI - [Observation of rare coexistence of the abdominal cavity organs surgical diseases]. PMID- 11036324 TI - [Observation of cardiac tumor in a child]. PMID- 11036325 TI - [Observation of the cutaneous melanoblastoma metastasis in small intestines]. PMID- 11036326 TI - [Twisting of omentum of the sigmoid intestine]. PMID- 11036327 TI - [The injury of the vaginal fornix penetrating into the abdominal cavity]. PMID- 11036328 TI - [The observation of incarcerated hernia in premature infant]. PMID- 11036329 TI - [Angiofollicular lymphoma of mesenteric localization in a child]. PMID- 11036330 TI - [Yes or no when it comes to antibiotics in acute otitis?]. PMID- 11036331 TI - [A rational use of antibiotics counteracts the development of resistance]. PMID- 11036332 TI - [Consensus on acute otitis in children: antibiotic treatment of uncomplicated otitis is questioned]. PMID- 11036333 TI - [Evidence-based medicine begins at home. Incentives for better decisions in the clinical everyday]. AB - Everyday clinical work raises many questions regarding diagnosis, prognosis and treatment. Traditional sources such as text books do not give all the answers, especially not in situations when decisions are needed without further delay. By learning the fundamentals of evidence-based medicine we may stand a better chance of founding our decisions on the best available evidence in order to provide better patient care. A focused question need to be posed, followed by a cost effective literature search. This clinical question comprises of four parts: Who is the patient? Which is the intervention? What is the comparison? What outcome is relevant to the patient and what are the adverse effects? PMID- 11036334 TI - [A conference report. Evidence-based medicine supported by qualitative research]. PMID- 11036335 TI - [To sail and navigate on the sea of medicine]. PMID- 11036336 TI - [Treatment of localized prostatic cancer--different alternatives. Patients with localized prostatic cancer must actively participate in the choice of therapy]. AB - The treatment options currently available for patients with clinically localized prostate cancer are radical prostatectomy, irradiation and deferred symptom guided treatment. Today we do not have any controlled data indicating that any of these treatments is better than the other. Management policies that have evolved are, at best, based on structured comparisons of data from uncontrolled treatment series. In such comparisons the differences between the various treatment options with respect to disease-specific survival up to 10 to 15 years after diagnosis are modest. These comparisons are based on patient series diagnosed 15 to 20 years ago, i.e. before prostate specific antigen (PSA) was available as a marker for prostate cancer. The utilization of PSA has dramatically changed the conditions for diagnosing prostate cancer, in that with PSA we find more men with prostate cancer with smaller tumors at diagnosis. Today we find many tumors because of an elevated PSA followed by systematic biopsies in men without palpable prostatic lesions. Thus it is difficult to translate long-term outcome data from older patient series to the current situation in which tumors are found under changed clinical conditions. Consequently, a patient can only be informed about the different treatment options and their side effects. In the treatment decision process he has to balance a possible benefit in survival against side effects, according to his own preferences. We need randomized studies comparing the different treatment options. Such research is under way in Scandinavia, but due to the long course of the disease we will probably have to wait many years before we can answer the questions which sparked these studies. PMID- 11036337 TI - [VLCD a safe and simple treatment of obesity]. AB - This review summarizes Swedish experience with VLCD (Very Low Calorie Diets). VLCD-treatment is a safe and relatively simple way to induce weight reduction in obese patients. The rapid and profound initial weight loss reduces cardiovascular risk factors and relieves obesity-associated symptoms. Weight loss on the order of 20-25 kg is common after 12-16 weeks of treatment. The long-term results, about 10% weight reduction after two years, are similar to what can be expected with pharmacological treatment. VLCD's should be incorporated into long-term treatment programs including diet, physical exercise and lifestyle modification. A team of nurses and/or dieticians can, to a large extent, manage a VLCD-program, restricting the need for involvement of the physician. PMID- 11036338 TI - [Low malignancy grade glioma in the Uppsala++/Orebro region. Prognostic factors and survival among 119 patients]. AB - The benefit of surgery and radiotherapy for patients with low-grade gliomas is controversial. We studied case records of 119 patients diagnosed with low-grade glioma during 1987-1993 at our hospital. Statistical analysis was made on overall survival and on survival related to histological diagnosis and treatment modality. A large variation in survival of patients with similar histological diagnosis was seen. No correlation was found between survival and treatment modality. This clinical diversity probably reflects variations in tumor biological characteristics of low-grade gliomas. Greater understanding of the biology of these tumors will thus be needed for more effective therapies. PMID- 11036339 TI - [Risk of increased malignancy a reason for follow-up of low malignancy grade glioma. Early identification of high-risk patients is the key to good care]. AB - Management of patients with low-grade gliomas remains a challenge. Controversies exist regarding the value of surgery and radiotherapy as well as the right timing of these treatments with respect to disease course. Several factors have been shown to have a positive correlation with better outcome, involving tumor size and localization, age and clinical condition of the patient on presentation, and epileptic seizures as a single symptom. The absence of contrast enhancement on CT scans is also a favorable prognostic factor. Taking into account the results of these studies as well as our own experience, we suggest some strategies for the management of patients with low-grade gliomas. The importance of identifying "high-risk" patients by systematic clinical and radiological follow-up is discussed. PMID- 11036340 TI - [Licorice--an old drug and currently a candy with metabolic effects]. PMID- 11036341 TI - [Charles Emil Hagdahl--physician, cookery books writer, art collector and reformer of correctional care]. PMID- 11036342 TI - [Mobile telephones and the risk of brain tumor--the principle of precaution should be practiced]. PMID- 11036343 TI - [A reply: the principle of precaution and mobile telephones--no reason to limit the use now]. PMID- 11036344 TI - [Pay attention to children with language development deviations]. PMID- 11036345 TI - [TSH and TPOAb should be the first to analyze in suspected dysfunction of thyroid gland]. PMID- 11036346 TI - [Mass screening for early discovery of colorectal cancer--are we ready]. PMID- 11036347 TI - [Final reply: the view on circumcision reveals an undemocratic attitude]. PMID- 11036348 TI - [Conclusive comment: our hopes are toward a more open religious attitude]. PMID- 11036349 TI - [Circumcision should and will be discussed]. PMID- 11036350 TI - [Circumcision of boys--a not-a-problem in which many are involved]. PMID- 11036351 TI - [Girls in Sweden are circumcized in spite of more stringent notification law]. PMID- 11036352 TI - [Memento mori--it's not possible to inform the death away!]. PMID- 11036353 TI - [Computer tomography or not in minor head injury? A study from the USA shows that it is possible to reduce the number of examinations]. PMID- 11036354 TI - [Glomerulonephritis--an occupational disease--new discoveries on a disease associated with exposure to toxic compounds]. PMID- 11036355 TI - [Only a directed prevention against meningococcal disease is justified. Inform patient's contacts about the risk of infection instead]. PMID- 11036356 TI - [Evaluation of analytic test methods. Stomach-intestines status is the best diagnostic method in suspected vitamin B12 deficiency]. AB - In the great majority of cases vitamin B12 deficiency is caused by chronic atrophic gastritis. This study has compared several different tests for the identification of patients for whom substitution therapy is needed or warranted. It is concluded that serum tests reflecting the state of the gastric body mucosa (serum pepsinogen A combined with serum gastrin or serum pepsinogen C) appear to be the logical first step in the workup of these patients in terms of high sensitivity and superior specificity as compared with other tests. PMID- 11036357 TI - [Increased procalcitonin level in bacteriogenic metabolic disturbances. A new possibility for diagnosis and treatment monitoring in sepsis]. AB - Earlier observations of increased plasma concentrations of immunoreactive calcitonin (32 amino acids) in sepsis and other non-tumorous conditions may be explained by increased secretion of procalcitonin, the 116-amino acid prohormone. At present, the site(s) of origin of procalcitonin in sepsis, the factors regulating its biosynthesis and release, the route(s) of its elimination from blood as well as its biological function(s) are unknown. The rapid increase in procalcitonin concentration in sepsis--in some patients earlier than that of C reactive protein--and decrease upon successful chemotherapy makes procalcitonin a potentially important biomarker in monitoring patients with suspected or confirmed sepsis. PMID- 11036358 TI - [Laboratory diagnosis of gastrinoma. Nordic collaboration can improve the quality]. AB - Measurement of gastrin in serum or plasma in patients with gastrinoma may be complicated by the presence of circulating biosynthetic intermediates which may not be detected by commonly available immunoassays. In contrast, the "processing independent analysis" of gastrins developed by professor Jens Rehfeld et al in Copenhagen detects gastrin forms irrespective of their size. The authors review gastrinoma pathophysiology, the biochemistry of gastrin and other biomarkers of gastrinoma, the differential diagnosis of hypergastrinemia as well as other methods currently employed in the workup of gastrinoma patients, and illustrate with a clinical case. PMID- 11036359 TI - [Acute abdomen calls for considerable care resources. Analysis of 3727 in patients in the county of Stockholm during the first quarter of 1995]. AB - A total of 3,727 in-patients with acute abdominal symptoms were identified during the first quarter of 1995 at the surgical clinics of the nine hospitals with emergency departments in the county of Stockholm. The diagnoses were: non specific abdominal pain 24%; cholecystitis 9%; appendicitis 8%; bowel obstruction 7%; intra-abdominal malignancy, diseases of the urinary tract and peptic ulcer 6% each; gastrointestinal hemorrhage, diverticulitis of the colon and pancreatitis 5% each; other diseases as a cause of abdominal symptoms, 19%. 1,601 operations were performed of which 47% were endoscopic procedures. The mean duration of hospital stay was 4.8 days. The length of stay increased significantly with age. The age-related relative frequency of hospitalization due to acute abdominal pain was also dramatically higher in the elderly cohorts. These facts and the prognosis of an 18% increase of inhabitants 50 years of age or older until 2010 in Greater Stockholm signal an increased need of hospital resources for this large group of patients in the coming years. PMID- 11036360 TI - [Incorrect use of drugs alarms physicians, nurses and pharmacists. Every other patient doesn't follow physician's prescription]. PMID- 11036361 TI - [Alarm from expert meeting when it comes to Swedish clinical research. "We are no longer as good as our reputation used to be"]. PMID- 11036362 TI - [IT development at the department of orthopedics in Skovde. Better patient information with "personal" computer printouts]. PMID- 11036363 TI - [Lombroso introduced a new perspective on forensic psychiatry: focus on the criminal, not the crime]. PMID- 11036364 TI - [When twilight sets in... Hello Lakarforbundet, where are you?!]. PMID- 11036365 TI - [Answer from the Lakarforbundet: we are aware of the problem--and we must find a solution together]. PMID- 11036366 TI - [In the light of the Adel-reform--physicians are the last piece in a jig-saw puzzle]. PMID- 11036367 TI - [A reply on cervix cancer screening: cervical cytological test doesn't fulfil the requirements of a good screening test]. PMID- 11036368 TI - [The National Board of Health and Welfare on Creutzfeldt-Jakob disease: physicians are to be notified about suspected cases]. PMID- 11036369 TI - [Risks and information, and the difference between understanding and the the ability to perform]. PMID- 11036370 TI - [HbA1C control also in patients with diabetes type 2!]. PMID- 11036371 TI - [Nurses' aides are not allowed to handle drugs--but there are some exceptions]. PMID- 11036372 TI - [Temperature measurements in health care--a question of quality assurance]. PMID- 11036373 TI - [Symptoms--always subjective!]. PMID- 11036374 TI - [Constructional weakness of Turbuhaler]. PMID- 11036375 TI - [Surgery and radiotherapy best in the treatment of rectal cancer. Correct use of patient registries can further increase the quality]. PMID- 11036376 TI - [More accurate tumor diagnosis results in more efficient treatment. New techniques makes the preoperative staging of gastrointestinal cancer possible]. PMID- 11036377 TI - [Improved therapeutic results in rectal cancer. Preoperative radiotherapy and continuing education of the surgeons is efficient]. AB - The aim of the Stockholm Colorectal Cancer Study Group is to improve treatment results in colorectal cancer. Between 1980 and 1993 the group conducted two randomised trials on preoperative radiotherapy in rectal cancer including 1406 patients. In both trials radiotherapy significantly reduced local recurrence and improved cancer specific survival. To further improve treatment results, four workshops have been held to introduce the "TME concept" to surgeons in Stockholm. The workshops included live surgical demonstrations and histopathology sessions. When patients included in the Stockholm trials (1980-1993) were compared to patients included in the "TME project" (1995-1996) the rate of abdominoperineal procedures was reduced from 60 to 27 per cent. Within two years of follow up the local recurrence rate was reduced from 15 to 6 per cent and cancer related deaths from 16 to 9 per cent. It is concluded that surgical teaching initiatives may have a significant impact on cancer outcomes. PMID- 11036378 TI - [Quality registry for better treatment of rectal cancer established. Economical support is required to use the benefits of the information]. AB - The treatment of rectal cancer has changed significantly during the last 30 years. With improved surgical technique and the introduction of preoperative radiotherapy sphincter preserving surgery is now predominant and the rate of local recurrence has been reduced substantially. However, new therapy concepts may also introduce an increased risk of complications. A register to monitor quality control in rectal cancer treatment in Sweden was established in 1995. It covers over 95 per cent of the patients with rectal cancer reported to the Swedish National Cancer Registry. Collection of data and validation are done by six regional oncology centres under supervision of surgeons appointed by the hospitals involved. The results are then collated to a nationwide quality register, enabling regions to compare themselves with other regions, and hospitals with other hospitals. PMID- 11036379 TI - [Regional medical care program in Stockholm-Gotland region: a first step toward a unified care of patients with stomach cancer]. AB - Cancer treatment in Sweden is coordinated by regional oncology centers responsible for keeping registers of cancer cases and assisting the medical profession in establishing regional consensus on the treatment of specific cancer forms. The article describes the development of a regional multidisciplinary protocol for gastric cancer, with standardization of preoperative investigations and curative surgery. A follow-up procedure deals with postoperative problems and defines recurrence rates. PMID- 11036380 TI - [Newly discovered human retroviruses. Association with disease is still undetermined]. AB - Retroviruses are enveloped RNA viruses which can transcribe RNA to DNA and integrate into the chromosomal DNA of their host cell. Heritable integrations give rise to endogenous retroviral sequences (ERVs). The rest is exogenous, infecting from individual to individual. This survey highlights an emerging scenario in human retrovirology. Humans have thousands of distinct ERVs. Although most are damaged by mutations, many are expressed as RNA, a few also as proteins and viral particles. The latter are not known to be infectious. Obviously, human ancestors encountered many different exogenous retroviruses, some of which may still be extant. In fact, an exogenous retrovirus related to ERVs was recently discovered. It is the fifth human exogenous retrovirus, human retrovirus 5 (HRV 5). It succeeds the two human T-lymphotropic viruses (HTLVs) and the two human immunodeficiency viruses (HIVs). The newly discovered endogenous and exogenous human retroviruses are now being investigated for association with disease. There are indications of selective ERV activation in multiple sclerosis, schizophrenia and seminoma. HRV-5 has been associated with rheumatoid arthritis, systemic lupus erythematosus and non-Hodgkin lymphoma. It is not yet known whether these first observations signal a pathogenic role for the newly discovered retroviruses. PMID- 11036381 TI - [The star-shaped cells. Astrocytes are involved in the pathogenesis and progress of neurological diseases]. AB - Recently, knowledge about the role of astrocytes in the brain has increased substantially. As a result we have had to rethink old views regarding how the brain works at the cellular level. Neurons can no longer be regarded as the only cell types of functional significance. The picture instead appears to be far more complex, with an ongoing exchange of information between different cell types, and this interaction is suggested to be particularly important between neurons and astrocytes. Astrocytes express receptors for different classes of neurotransmitters, and have both voltage and receptor operated ion channels. Through active uptake and release of ions, neurotransmitters and water they control the brain interstitium. Intercellular communication via transfer of neuroactive substances through gap junctions makes it possible to coordinate different activities in large areas of the brain. Dysfunction of astrocytic physiology is thought to contribute to the pathogenesis and progress of various neurological disorders such as epilepsy, stroke and cerebral edema. PMID- 11036382 TI - [Nasal polyposis--of interest to several specialties. Improved diagnosis and new therapeutic possibilities]. AB - Nasal polyposis, a disease seen mainly in humans and dominated in most cases by an eosinophilic inflammation of unknown etiology, has been an enigma for a few thousand years. Polyps are, however, overrepresented in patients with general airway inflammation as seen in asthma and idiopathic rhinitis, and in patients with infective nasal disease of chronic origin as seen in cystic fibrosis, immune deficiencies and dental sinusitis. Due to the eosinophilic nature of the inflammation, polyps are on the whole sensitive to local steroid treatment with spray or drops. Surgery is indicated when the polyps are solitary or when medical treatment fails. PMID- 11036383 TI - [Nasal polyps from a patient's perspective. Good results with drug therapy and surgery]. PMID- 11036384 TI - [Nasal congestion among women. At least every fifth pregnant woman suffers--most common among smokers]. PMID- 11036385 TI - [Reduced but better health care for armed forces]. AB - The Swedish Armed Forces are presently undergoing the most extensive reorganization and downscaling in their history. In a new set of objectives for the Swedish Armed Forces Medical Service laid down by the Surgeon General, the aim is to provide combat casualty care with a quality on par with that of the civilian, peace-time health care system. This will be achieved by establishing advanced trauma care by specialist physicians and nurses closer to the point of injury, using armoured medical evacuation vehicles, and by introducing new treatment modalities such as intraosseous infusion using hyperoncotic solutions. PMID- 11036386 TI - [Narrative medicine--a story about a patient and his disease]. PMID- 11036387 TI - [About back pain--in the body and/or in the soul. A comment to a SBU-report on back pain]. PMID- 11036388 TI - [Treatment of depression in primary health care. Competence, engagement and empathy give results]. PMID- 11036389 TI - [Broader perspective is required]. PMID- 11036390 TI - [Chronic fatigue syndrome. Different definitions are confusing]. PMID- 11036391 TI - [What is the cause of low concentration of B12 in cerebrovascular fluid in multiple sclerosis?]. PMID- 11036392 TI - [Language and speech--not a subject for a state of the art conference on child health care]. PMID- 11036393 TI - [What does a human life cost?]. PMID- 11036394 TI - [On the dialogue with theologians]. PMID- 11036395 TI - [Radiation and radiation protection in the society]. PMID- 11036396 TI - The incidence of opportunistic fungi in patients suspected of tuberculosis. AB - The incidence of opportunistic fungi in bronchoalveolar lavage specimens from patients suspected of tuberculosis in Isfahan, Iran, was determined. From 200 patients 36 yeasts (18%) and seven filamentous fungi (3.5%) were isolated. Out of 44 patients who had fungal infections, 12 cases were affected with definite tuberculosis. PMID- 11036397 TI - Changes of virulence factors accompanying the phenomenon of induced fluconazole resistance in Candida albicans. AB - We investigated a fluconazole-sensitive (MICflu = 5 micrograms ml-1) clinical isolate and a fluconazole-resistant (MICflu > 80 micrograms ml-1) laboratory mutant Candida albicans strain developed from the sensitive one. We studied putative virulence factors including germination, adherence ability to either buccal epithelial cells or acrylate surface, the secreted aspartic proteinase, and the extracellular phospholipase activity of the two strains as well as their growth. The fluconazole-resistant strain proved to be superior to the original strain in all the virulence traits tested. The higher virulence of the fluconazole-resistant strain was also supported by a mouse model. These results suggest that the development of fluconazole resistance can be accompanied by serious morphological and physiological changes: several putative virulence traits, moreover the in vivo virulence can increase simultaneously. PMID- 11036398 TI - Scopulariopsis brevicaulis: a keratinophilic or a keratinolytic fungus? AB - The morphologic expression of human hair and nail invasion in vitro by Scopulariopsis brevicaulis isolates was studied by light microscopy on whole material and on semi-thin sections, and also by scanning and transmission electron microscopy. Only three isolates of the nine that were examined were keratinolytic, capable of both attacking keratinic substrates and demolishing their keratin. The process showed all the characteristic of enzymatic digestion and was in agreement with the biochemical and morphogenetic scheme proposed for other keratinolytic fungi during their invasion of human hair in vitro. All the active isolates were capable of developing structures related to surface erosion and radial penetration contemporaneously. However the extent and rate of keratinolysis were rather low when compared with the efficiency of other keratinolytic fungi. This finding suggests that S. brevicaulis is of secondary importance in the mineralization of keratinic substrates in natural environments. From the medical standpoint the mere demonstration of keratinolytic activity means that it may be regarded as a real cause of primary infection. PMID- 11036399 TI - Comparison of Etest with the broth microdilution method in susceptibility testing of yeast isolates against four antifungals. AB - A comparative evaluation of the Etest and the broth microdilution methods for antifungal susceptibility testing of 102 clinical yeast isolates against amphotericin B, fluconazole, itraconazole, and ketoconazole was conducted. The agreements between the Etest and the broth microdilution methods were 93.1% for amphotericin B 85.2% for ketoconazole, 82.3% for itraconazole and 79.4% for fluconazole. These results suggest that the Etest approach to antifungal susceptibility testing may be a viable alternative to the NCCLS reference methods for testing yeasts, but that further evaluations are needed. PMID- 11036400 TI - Anti-Cryptococcus activity of combination of extracts of Cassia alata and Ocimum sanctum. AB - The paper reports the anti-Cryptococcus activity of combination of ethanolic extracts of leaves of Cassia alata and Ocimum sanctum. The activity of combination of the extracts was heat-stable and worked at acidic pH. PMID- 11036401 TI - Oral yeasts and coliforms in HIV-infected individuals in Hong Kong. AB - The objective was to determine the oral carriage patterns of yeasts and coliforms and their relationships, if any, with age, risk group, CDC classification, CD4+ count and medications in a predominantly Chinese, HIV-infected cohort in Hong Kong. A prospective longitudinal study was carried out over a 12-month period, of 32 predominantly Chinese male HIV-infected cohort in a hospital setting in Hong Kong. Oral carriage rates were determined by the concentrated rinse culture method and correlated with other clinical parameters using regression analysis. A total of 73 oral rinse samples were collected and the weighted mean carriage rates of oral yeasts and coliforms were 54.8% and 28.8%, respectively. The most common yeast and the Enterobacteriaceae isolated were Candida albicans and Enterobacter cloacae, respectively. An increased carriage rate of yeasts was associated with zidovudine usage and Centres for Disease Control (CDC) stage IV of the HIV infection whereas the opposite was associated with the usage of antiparasitics and multivitamins. Although the oral carriage rate of coliforms was significantly lower in individuals taking antibacterials and multivitamins, it was not significantly influenced by age, CD4+ lymphocyte count and the intake of antivirals, antifungals or folates. These data imply that oral yeast carriage in HIV infection is related to the severity of the disease as opposed to oral coliform carriage which appears to be unusually transient in the study cohort. PMID- 11036402 TI - In vitro susceptibilities of 11 clinical isolates of Exophiala species to six antifungal drugs. AB - The antifungal activities of miconazole, terbinafine, itraconazole, UR 9825, voriconazole and amphotericin B against 11 clinical isolates of Exophiala spp. were tested by the broth microdilution method. All drugs were very active against Exophiala spp.. The 90% minimal inhibitory concentration (MIC90) ranged from 0.125 to 1 microgram ml-1. Terbinafine was the most active drug against Exophiala spinifera, Exophiala dermatitidis and Exophiala castellanii and seems to be a promising agent in the treatment of infections caused by these fungi. PMID- 11036403 TI - Case report. Chronic necrotizing pulmonary aspergillosis in a non-neutropenic patient treated with liposomal amphotericin B. AB - Chronic necrotizing pulmonary aspergillosis is a rare infection encountered mainly in immunocompromised patients. We present the case of a young woman where the diagnosis of chronic necrotizing pulmonary aspergillosis was established after an open biopsy of the right lung. Liposomal amphotericin B was administered postoperatively for 10 days with moderate success. In a concise review we summarize the basic notions of chronic necrotizing pulmonary aspergillosis. PMID- 11036404 TI - Case report: chronic dermatophyte infection in a patient with vitiligo and discoid lupus erythematosus. AB - We report the case of a patient suffering from vitiligo and discoid lupus erythematosus. The nails of the left hand were involved with subungual hyperkeratosis. He was treated by local and systemic corticosteroid and chloroquine but hyperkeratotic lesions progressed on the affected sites and were found to be caused by Trichophyton violaceum. PMID- 11036405 TI - [Methodology of the evaluation and the reproductive potential of girls]. AB - Health status of girls as future mothers causes great apprehension. The term "reproductive potential" is recommended to be used instead of "reproductive health". A program for evaluating the reproductive potential has been developed, including 5 sections. Results of evaluating the reproductive potential of 12,272 girls in St. Petersburg, Yakutsk, Novgorod, and the Novgorod region are analyzed. PMID- 11036406 TI - [Dynamics of the cause of death of the Saratov region population]. AB - Short-term fluctuations in mortality are analyzed for the Saratov region in order to indirectly evaluate the efficiency of public health. Analysis revealed new tendencies in population deaths, which however manifested not so clearly as was expected. Studies and prediction of short-term fluctuations in the mortality coefficient from causes of death remain an important problem. PMID- 11036407 TI - [Prevalence of congenital malformations]. AB - Analyzes the incidence of congenital malformations in children and the relationship between hereditary diseases and average life span for the city of Krasnodar in 1987-1998, with special emphasis on morbidity, death/birth rate, and neonatal mortality caused by hereditary diseases. The incidence of hereditary diseases incompatible with life has increased in recent years and is expected to increase more, due to deterioration of social and economic situation in the country, communal conditions, and ecology. PMID- 11036408 TI - [Opinion of physicians concerning sexual health of children]. AB - Opinions of physicians on sexual health of prepubertal children were studied using questionnaires, which were distributed among pediatrists, endocrinologists, gynecologists, neurologists, psychotherapists, urologists, sexologists, dermatologists, and specialists in sexually-transmitted diseases. The majority of respondents of all specializations could not give definitions of sexual health, its criteria and pediatric sexopathology. Almost all considered desirable the introduction of a pediatric sexologist at health centers and regular sexual education of prepubertal children. PMID- 11036409 TI - [Social and hygienic characteristics of families of schoolchildren with chronic diseases]. AB - Comparative analysis of families of schoolchildren in Vladikavkaz demonstrated the social and hygienic characteristics of families with schoolchildren suffering from chronic diseases, necessitating the formation of a special priority group with a complex of specific medical and social requirements, in order to develop prophylactic recommendations. The role of a family in prevention of diseases of schoolchildren should be increased and special recommendations should be developed for families with schoolchildren suffering from chronic diseases. PMID- 11036410 TI - [Problems in meeting the requirements for health services]. PMID- 11036411 TI - [Analysis of the flow of users of health services at a diagnostic center]. AB - A marketing investigation was carried out in 1998 in the Yekaterinburg Multiprofile Diagnostic Center in order to detect the problems in the activity of this institution and adopt the managing decisions. A random sampling of 405 subjects was selected, 58.8% of them were examined at the expense of budget financing and 41.2% for extra payment. The results indicate that 50.5% patients consulting the specialists of the Center were in need of diagnosis, 26.2% needed a more accurate diagnosis of a previously diagnosed condition, 9.4% patients consulted for correcting their treatment protocols, 8.4% were followed up at the Center, and 4.9% were examined within the framework of prophylactic check-ups. Half of patients were completely satisfied, about one-third were satisfied partially, and only 2% were not satisfied at the Center. 86.9% respondents aid that they would apply into the Center again, if necessary. The main sources of information of the city and region population were local physicians (52.7%), friends and relatives previously examined at the Center (19.8%). Marketing investigations in public health become an integral part of effective management under conditions of expanding the scope of marketing services in practical medicine. PMID- 11036412 TI - [Integration of activities of regional hospitals and territorial medical institutions]. AB - Medical and economic efficiency of regional therapeutic and prophylactic institutions is to develop in integration with therapeutic and prophylactic institutions of administrative territories of a subject of the federation, which necessitates modifications in the functions and organizational structure of organization and methodology departments of regional, central, and municipal hospitals. PMID- 11036413 TI - [Current problems of rural public health reform]. AB - Main directions of the rural public health improvement are outlined and substantiated: creation of a regional network of medical institutions with integration of territorial public health systems, development of hospital replacing technologies which can be realized in the rural regions, transfer to the general practitioner model with rational distribution of functions between specialists, medics and paramedics, and introduction of automated managing systems. PMID- 11036414 TI - [Organizational aspects in creation of the register of patients with renal insufficiency]. AB - The authors validate the need in creation of regional registers of patients as exemplified by the register of patients with renal failure in the Republic of Tatarstan. Philosophy and stages in the formation of the registers are suggested, making use of computer information technology "Dialysis Center". The technology is characterized in detail with regard to public health organization and management at a regional level. PMID- 11036415 TI - [Development of hospital-substituting forms of medical care in the Samara region]. AB - Development of hospital-substituting forms of medical care in the Samara region is discussed, 13% of patients treated at hospitals were rendered medical care through hospital-substituting institutions, which resulted in economy of 137 million roubles. The authors make their suggestions on optimizing statistical forms of files and records to be used in registration of medical care at day-time hospitals. PMID- 11036416 TI - [Interterritorial accounts for medical care in 1993-1999]. PMID- 11036417 TI - [Teaching of history of medicine at the St. Petersburg Medical University]. AB - Training in history of medicine is carried out at St. Petersburg I. P. Pavlov State Medical University in the context of all forms of human society: "nonprogressive", "cyclic development", and "progressive". Each topic is presented in logical sequence with consideration for factors of mutual effect and mutual contributions of medicine of different regions, including Russia. Major attention is paid to the key points in development of medicine, its regularities, and basic characteristics. PMID- 11036418 TI - [Development of clinical training during the 1st half of the 19th century. I]. PMID- 11036419 TI - [History of the organization of the Medical Institute at the Moscow University]. PMID- 11036420 TI - [Role of the nobility in the development of zemstvo medicine in the Moscow province]. PMID- 11036421 TI - [Contributions of D.N. Zhbankov to the development of zemstvo medicine]. PMID- 11036422 TI - [Data on medical support of war actions of frontier troops during the 1st period of the Great Patriotic War]. PMID- 11036423 TI - [Medicine in medieval Russia. 2]. PMID- 11036424 TI - Patients with tuberculosis in Bolivia: why do they die? AB - The objective of this research was to analyze why patients with tuberculosis (TB) die and to evaluate whether there are factors contributing to their fatal outcome that could be corrected. A cross-sectional observational study was conducted of the patients with active TB or its sequelae admitted to the TB ward of the main public hospital in the city of Santa Cruz, Bolivia, over a 29-month period, from October 1993 through February 1996. The available records of the patients who died during hospitalization were reviewed. Out of 597 patients, 94 of them (15.7%) died. We examined the records of 90 of these 94 patients. Their mean age was 35.1 years (standard deviation, 16.7 years), and 45 of the patients (50.0%) were male. On admission 42 of the 90 patients (46.7%) had never been treated for TB or had received anti-TB treatment for less than one month, 23 (25.6%) had returned after having abandoned their TB treatment, 8 (8.9%) had had an erroneous diagnosis, 6 (6.7%) had tuberculosis sequelae, 6 (6.7%) were undergoing tuberculosis treatment, and 5 (5.6%) were known to have multidrug-resistant TB. Of the 90 patients, 83 (92.2%) had pulmonary tuberculosis (median lobes affected, 4), 6 (6.7%) had pleural tuberculosis, and 12 (13.3%) had extrapulmonary tuberculosis (some patients had more than one form of TB). Patients died a median of 5.5 days after entering the TB ward. The causes of death were: hemoptysis, 6 patients (6.7%); other tuberculosis-related causes, 65 patients (72.2%); drug reactions, 6 patients (6.7%); nontuberculosis causes, 6 patients (6.7%); and undetermined causes, 7 patients (7.8%). Factors possibly contributing to death were late diagnosis (38.9%), errors in follow-up (14.4%), and errors in treatment (24.4%). In conclusion, most patients with active or inactive TB admitted to our ward died as a consequence of tuberculosis. There were several factors possibly contributing to their fatal outcome that could be corrected. PMID- 11036425 TI - [Influence of helminthic infections and nutritional status on immune response in Venezuelan children]. AB - We investigated the influence of nutritional status, as determined from anthropometric measurement, and of helminthic infections on the immune response of children of low socioeconomic status in two rural communities in Venezuela: El Cardon in the state of Nueva Esparta and San Daniel in the state of Miranda. A total of 125 boys and girls between 2 and 15 years old participated in the study. Their socioeconomic stratum was determined by a modified Graffar method. A physical examination was performed, as was also an anthropometric evaluation that took into account three indicators--weight-for-height, weight-for-age, and height for-age--according to parameters established by the World Health Organization. Other examinations included feces, secretory IgA in saliva, total serum IgE, and anti-Ascaris-specific immunoglobulins. The children in both of the communities were in strata IV and V of the of Graffar scale, with a significantly greater number of stratum V inhabitants in San Daniel (P < 0.001). The results suggest that exposure level and individual susceptibility to the parasites are determining factors in parasitic infection and immune system behavior. The intensity of the parasitic burden plays an important role in stimulating polyclonal IgE, which diminishes the effectiveness of the specific response to those infections. On the other hand, nutritional deficiencies could change the immune mechanisms of the mucous membranes, negatively influence the synthesis of secretory IgA, and stimulate the production of polyclonal IgE. Poor sanitary and socioeconomic conditions promote more exposure to gastrointestinal parasites and a deficient nutritional status, which modulates the immune response and affects serum IgE and secretory IgA production mechanisms. PMID- 11036427 TI - Microbial quality of water in rural communities of Trinidad. AB - A cross-sectional study was conducted in four rural communities of northeastern Trinidad to determine the microbial quality of water supply to households and that quality's relationship to source and storage device. Of the 167 household water samples tested, total coliforms were detected in 132 of the samples (79.0%), fecal coliforms in 102 (61.1%), and E. coli in 111 (66.5%). There were significant differences among the towns in the proportion of the samples contaminated with coliforms (P < 0.001) and E. coli (P < 0.001). Of 253 strains of E. coli studied, 4 (1.6%) were mucoid, 9 (3.6%) were hemolytic, and 37 (14.6%) were nonsorbitol fermenters. Of 69 isolates of E. coli tested, 10 (14.5%) were verocytotoxigenic. Twenty-eight (14.0%) of 200 E. coli isolates tested belonged to enteropathogenic serogroups. Standpipe, the most common water source, was utilized by 57 (34.1%) of the 167 households. Treated water (pipeborne in homes, standpipes, or truckborne) was supplied to 119 households (71.3%), while 48 households (28.7%) used water from untreated sources (rain, river/stream, or well) as their primary water supply. The type of household storage device was associated with coliform contamination. Water stored in drums, barrels, or buckets was more likely to harbor fecal coliforms (74.2% of samples) than was water stored in tanks (53.3% of samples), even after controlling for water source (P = 0.04). Compared with water from other sources, water piped into homes was significantly less likely to be contaminated with total coliforms (56.9% versus 88.8%, P < 0.001) and fecal coliforms (41.2% versus 69.8%, P < 0.01), even when the type of storage device was taken into account. However, fecal contamination was not associated with whether the water came from a treated or untreated source. We concluded that the drinking water in rural communities in Trinidad was grossly unfit for human consumption, due both to contamination of various water sources and during household water storage. PMID- 11036426 TI - [Infant mortality in Rio de Janeiro, Brazil: risk areas and distance traveled by patients to get to health care facilities]. AB - The infant mortality rate has been considered a summary of the quality of life and level of development of a given population. However, this indicator is very sensitive to such simple measures as oral rehydration therapy, vaccination, and continuation of breast-feeding. Given that such health activities have become more widespread, an infant mortality rate may no longer reflect a particular development model. With the aim of broadening the discussion regarding infant mortality, this study analyzed the 153 neighborhoods of the city of Rio de Janeiro, Brazil. Our objective was to identify areas with more risk, and the priority groups for interventions to decrease infant mortality. We analyzed neonatal and postneonatal mortality in each neighborhood. We also identified the children's home neighborhood and the location of their deaths and related these results to the socioeconomic classification of the corresponding neighborhoods. In relation to the average infant mortality rate for the city, we could not make statistically significant comparisons for some neighborhoods due to their small number of births. One-third of the infant deaths could have been prevented with early diagnosis and treatment. Only 15% of the deaths were considered unavoidable. Both neonatal mortality and postneonatal mortality were geographically dispersed, with no direct association with the socioeconomic profile of the neighborhoods. An analysis of the children's place of residence and the location of their deaths showed flows of patients from poor areas to more affluent city areas with better health services. This pattern highlights the effect of access to quality medical care on infant mortality. PMID- 11036428 TI - [Rapid assessment of the impact of Haemophilus influenzae vaccine serotype b in Colombia. Public Health Laboratories]. AB - In May 1998 the Ministry of Health of Colombia started a universal vaccination campaign against Haemophilus influenzae type b (Hib) for children under one year of age. The impact of this intervention on the incidence of acute bacterial meningitis was assessed in 1999, using data from the laboratory-based surveillance system coordinated since 1994 by the Microbiology Group of the Colombian National Institute of Health. The analysis compared the annual number of cases of Hib meningitis in children under one year of age diagnosed through the surveillance system before the vaccine was introduced with the number of cases reported during the first year after the vaccine's introduction. The expected number of cases, given the average annual number of cases diagnosed between June 1994 and June 1998, was compared with the number of cases observed after the vaccination program was introduced, from June 1998 through May 1999. To control for the quality of the surveillance system, a similar analysis was done for cases of meningitis due to Streptococcus pneumoniae. The analysis was restricted to those departments of Colombia that had consistently participated in the surveillance system. For the years 1994 through 1998 the numbers of confirmed cases of Hib meningitis were, respectively, 45, 37, 61, 64, and 31. In the period after the vaccine's introduction 31 cases were observed, as compared to the 52 expected (P < 0.001). During the same annual periods there were 32, 26, 43, 48, and 42 confirmed cases of meningitis from S. pneumoniae in children less than 5 years old, showing no significant reduction in the expected number of those cases. The 40% decrease noted in Hib meningitis cases was not attributable to changes in the surveillance system and was due mainly to the effects of the vaccination program. PMID- 11036429 TI - [Impact of Streptococcus pneumoniae on pneumonia in Latin American children. SIREVA-Vigia Group]. AB - Community-acquired pneumonia is one of the leading causes of infant morbidity and mortality. Studies conducted in developing countries indicate that the most serious symptoms of pneumonia are associated with bacterial causes, mainly Streptococcus pneumoniae, followed by Haemophilus influenzae type b. Managing those infections in children under two years of age is hindered by the lack of appropriate vaccines and by the decreased susceptibility of S. pneumoniae to penicillin and other antibiotics. In 1993, at the initiative of the Regional System for Vaccines of the Pan American Health Organization, and with funding from the Canadian International Development Agency, a study was designed to identify the S. pneumoniae capsular types that cause invasive disease in Latin American children under 5 years of age. The objective of the study was to determine the ideal composition of a conjugate vaccine that could be used in Latin America, and the penicillin susceptibility of the S. pneumoniae isolates. The initiative was undertaken in Argentina, Brazil, Chile, Colombia, Mexico, and Uruguay. This report analyzes the information that the participating countries generated on pneumococcal pneumonia. A total of 3,393 children were found with systemic S. pneumoniae infections, of which 1,578 corresponded to pneumonias. The analysis focused on 1,409 cases of pneumonia in Argentina, Brazil, Colombia, Mexico, and Uruguay. Of the children, 63.8% of them were under two years of age. Twelve prevalent capsular types were identified, of which serotypes 14, 5, and 1 were the three most common in the majority of the countries. At the beginning of the study the highest level of penicillin resistance was found in Mexico (47.0%), and the lowest in Colombia (12.1%). Over the 1993-1998 period, resistance to penicillin increased in the five countries. Penicillin resistance was associated with a small number of capsular serotypes, mainly 14 and 23F. The first of those serotypes was resistant to penicillin and to trimethoprim-sulfamethoxazole, and the second was multiresistant. The frequency of resistance to trimethoprim sulfamethoxazole was high in all of the countries; Argentina had the highest level, 58.0%. A decrease in susceptibility to chloramphenicol was uncommon, except in Colombia, where there was a resistance level of 23.4%. Resistance to erythromycin was low in all the countries, and all the isolates were susceptible to vancomycin. PMID- 11036430 TI - Economic burden of illness from pesticide poisonings in highland Ecuador. AB - Active surveillance of acute pesticide poisonings in a potato-growing region of highland Ecuador during 1991-1992 uncovered a rate of 171/100,000, due predominantly to occupational exposures to organophosphate and carbamate pesticides. Occupational exposure among agricultural workers was the most common reason for poisoning (32 male workers and 1 female worker, out of a total of 50 cases). Of these 33 cases, 28 of them reported pesticide application as the work task just prior to poisoning, with over 80% citing the use of World Health Organization Hazard Category I pesticides. The suicide rate of 17.1/100,000 and the overall mortality rate of 20.5/100,000 that we found are among the highest reported anywhere in the world. At the exchange rates prevailing at that time, median costs associated with these poisonings were estimated as follows: public and social security health care direct costs of US$ 9.85/case; private health costs of US$ 8.33/case; and lost-time indirect costs of US$ 8.33/agricultural worker. Each one of those costs was over five times the daily agricultural wage, which was then about US$ 1.50. Further costing of pesticide poisonings should be carried out in other settings to provide appropriate information for decisions about pesticide use. In addition, integrated pest management should be further evaluated as an appropriate technology to reduce the economic burden of illness from pesticide poisonings in developing countries. PMID- 11036431 TI - [Health care reform and changes in nursing practice in philanthropic hospitals in Ribeirao Preto (SP), Brazil]. AB - This paper describes part of a multicenter study sponsored by the Pan American Health Organization to assess health care reforms and their implications for nursing in several countries. The objective of this research was to learn the views of nurses working in philanthropic hospitals in Ribeirao Preto, in the state of Sao Paulo, Brazil, regarding the changes in nursing practice coming from Brazil's health care reform and implementation of the Unified Health System (UHS). Data were obtained through structured interviews with seven nurses who met the selection criteria, from the three philanthropic hospitals in Ribeirao Preto. The nurses reported a decline in the quality of care and in the number of beds for UHS patients. The nurses reported that UHS implementation initially led to infrastructure improvements in the philanthropic hospitals. However, the reforms eventually shifted toward improving the care of private and privately insured patients. In addition, the nurses emphasized their heavy work loads and low pay. The nurses' reports indicated that Brazil's UHS is going through a crisis. In general, the nurses linked this crisis to problems in funding and allocation of resources. PMID- 11036432 TI - [Individualized drug therapy--need for improvement]. AB - The lack of dose adjustment to the individual needs of a patient is a major cause for the substantial difference between the efficacy of therapies established in large trials and the therapeutic results obtained in everyday practice (reduced effectiveness). In addition, these differences affect drug safety and ultimately also costs. Hence strategies to maintain good compliance and to avoid incompatibilities, drug interactions and adverse events should be applied whenever drugs are prescribed. This supplement compiles different techniques and measures which may support health professionals in reaching these goals. PMID- 11036433 TI - [Drug information resources in Internet]. AB - Many reliable drug information resources are now available free via the internet. The websites selected for this synopsis are apt to help the busy practitioner in his or her decision on an individualised drug therapy. PMID- 11036434 TI - [Individualization of compliance]. AB - Almost half of all prescribed drugs is taken by patients in an incorrect way, in a deficient way or not at all. Reasons for such an insufficient compliance behavior are multifaceted. In order to optimize patients' compliance, an individualized approach is necessary which requires both medical intervention and preventive measures. In the present article tools are described that may assist the physician in optimizing individual compliance. Such strategies are information, communication and verbal contract, treatment plan, auxiliary aids and follow-up appointments. PMID- 11036435 TI - [Consideration of drug absorption in customizing drug therapy]. AB - The rate and extent of drug absorption from the small intestine are related to the release of the active ingredient from a dosage form, its solubility in the liquid phase of gastrointestinal contents, and the transport of the dissolved compound or the intact dosage form from the stomach into the duodenum. With pharmaceutical preparations releasing the active compound within the stomach, and enteric-coated "micro"-formulations (micropellets), gastric emptying is possible during the interdigestive and the digestive period. Potential differences of drug absorption between fasting administration and intake during the digestive period are unpredictable, because they are related to the release characteristics of the dosage form. However, larger enteric-coated preparations like tablets can leave the stomach only with a phase 3 contraction of fasting motility; intake during the digestive period will result in gastric retention of this type of dosage form until all food has left the stomach and fasting motility is restored. Consequently the onset of drug absorption is delayed. This interaction between food and large enteric-coated dosage forms is predictable from pyloric function in relation to the gastric motility. As it occurs regularly, it can be taken into account when prescribing enteric-coated dosage forms. If concomitant intake of food and enteric-coated drugs is unavoidable, but a rapid onset of drug absorption is necessary, micropellets are the dosage form of choice. When the therapeutic effect is insufficient, drug dosage form and timing of drug administration should be checked before prescribing a different active compound. PMID- 11036436 TI - [Drug interactions in practice]. AB - As potential modulators of drug safety and effectiveness the compatibility and stability of drugs are important elements in drug prescription and drug administration to patients. According to various characteristics, important differences between areas of stationary hospital care and ambulatory care are evident. While the challenges in the ambulatory care setting have been focussed on dermatological topical medication, incompatibility assessment is a challenge for the physician and the pharmacist in a hospital setting. Particularly in intensive care patients often up to 20 different drugs are administered in only few infusion devices or through a limited number of parenteral infusion lines. Among the compounds most often incriminated to cause incompatibilities are furosemide, phenytoin, midazolam and diazepam when used as i.v. admixtures. In recent years new challenges arose in ambulatory care in the area of oncologic treatment. Cytotoxic drug therapy and supportive care therefore requires increasing attention in terms of incompatibility of ready to use combined drug admixtures. To prevent incompatibilities never more than one drug should be added, nutrition solutions should never be used as carrier solutions for drug administration, mixtures should not be stored and should be administered immediately after preparation. During the administration of unavoidable mixtures visible changes of the solutions should be carefully sought. PMID- 11036437 TI - [Individualization of drug therapy in renal or liver insufficiency]. AB - Individualisation of drug dosage in patients with renal or hepatic failure may prevent excessive drug accumulation and thus potentially reduce adverse drug reactions and costs. In renal failure, renal function may be estimated by combined evaluation of serum creatinine values and patient characteristics. Then individual elimination capacity of a given drug in the individual patient may be calculated and dosage accordingly adjusted. In severe liver cirrhosis, after peroral administration of drugs with a high extraction ratio each single dose has to be reduced because of increased bioavailability and decreased clearance. After i.v. administration and dosing of drugs with a low extraction ratio maintenance dose should be reduced either by prolonging the dosing interval or by decreasing each single dose. PMID- 11036438 TI - [Clinical relevance of drug metabolism polymorphisms]. AB - Individual variation in drug response is a substantial clinical problem. Research in pharmacogenetics is currently evolving in two directions, firstly to identify genes and gene products associated with certain diseases, which may serve as targets for new drugs, and secondly to identify genes and allelic variants of genes that affect the expected and adverse response to current and future drugs. In that respect drug metabolising enzymes play a key role. A readily and widely available pheno- and/or genotyping service for drug metabolism polymorphisms is currently not established and the clinical relevance has only been shown for a limited number of drugs. The overall pharmacologic effects are typically not monogenic traits, they are a result of an interaction of several genes encoding proteins involved in drug metabolism, disposition, and effects. Each individual represents a unique combination of polymorphic genes that are known to be involved in the metabolism and disposition of medications, in the target structures of drug therapy, and in the pathogenesis of diseases. This will lead to the development of DNA chips for genotyping which will guide the selection and dosing of drug therapy in the future. PMID- 11036439 TI - [Clinically relevant adverse drug interactions]. AB - Drug interactions may lead to adverse drug effects or therapeutic failure. Many clinically relevant unwanted interactions are caused by a change in the activity of cytochrome P450 isoenzymes or the activity of active drug transport systems (e.g. p-glycoprotein). Most drug interactions may be anticipated and prevented by dose modification or using alternative drugs. PMID- 11036440 TI - [Management of adverse drug effects]. AB - Adverse drug reactions (ADRs) are still considered one of the main problems of drug therapy. ADRs are associated with considerable morbidity, mortality, decreased compliance and therapeutic success as well as high direct and indirect medical costs. Several considerations have to come into play when managing a potential ADR. It is critical to establish an accurate clinical diagnosis of the adverse event. Combining information about drug exposure together with considering other possible causes of the reaction is crucial to establish a causal relationship between the reaction and the suspected drug. Identification of the underlying pathogenesis of an ADR together with the severity of the reaction will have profound implications on continuation of drug therapy after an ADR. Since spontaneous reports about ADRs are a key stone of a functioning post marketing surveillance system and therefore play a key role in improving drug safety, health care professionals are highly encouraged to report ADRs to a local or national organization. However, because the majority of ADRs is dose-dependent and therefore preventable, individualization of pharmacotherapy may have a major impact on reducing such events. PMID- 11036441 TI - [Investigation of patients with intracranial disease]. PMID- 11036442 TI - [Carotid stenosis is not treated sufficiently in Denmark]. PMID- 11036443 TI - [Carotid endarterectomy for symptomatic stenosis of carotid artery]. PMID- 11036444 TI - [Treatment of carotid stenoses--surgery or with balloon]. PMID- 11036445 TI - [Investigation of patients with transient cerebral ischemia]. PMID- 11036446 TI - [Acute glaucoma following endarterectomy of internal carotid artery]. AB - A 71 year-old woman had 85% and 60% stenoses of the right and left internal carotid arteries, respectively. Right-sided endarterectomy was performed because of amaurosis fugax. Two days postoperatively, she developed classical symptoms and signs of acute glaucoma. Right-sided acute angle closure glaucoma was diagnosed by tonometry and gonioscopy, and treated with laser-iridectomy. The patient was discharged two days later without neurological or ophthalmological deficits. Manifest acute glaucoma postoperatively seems never to have been reported. However, the choroid is not autoregulated, and the intraocular pressure has been reported to increase after internal carotid-endarterectomy. PMID- 11036447 TI - [Chronic subdural hematoma with apoplectic form or other non-characteristic onset]. AB - The medical records of all patients with the diagnosis subdural haematoma admitted to Gentofte county hospital during the period 1994-1998 were examined. In the five year period, 45 patients, (23 female and 22 male, mean age 74 years), were discharged from Gentofte county hospital, with the CT-verified diagnosis of subdural haematoma. The 45 patients were referred to hospital with the following diagnosis: Subdural haematoma, 19 pt's (42%): stroke/Transient ischaemic attack (TIA), 17 pt's (38%) and other, 9 pt's (20%). After clinical evaluation at the hospital the diagnoses were: Subdural haematoma 30 pt's (65%); stroke/TIA 9 pt's (20%) and other; 7 pt's (15%). In the group of 15 misdiagnosed patients, the correct diagnosis was established by CT-scan with a median delay of two days (range 0-49 days). Of the 15 patients, 14 were operated. One patient died, one had severe neurological defects. In conclusion, more than half of the patients with subdural haematoma were admitted with a diagnosis other than subdural haematoma, mainly stroke/TIA. In hospitals receiving patients with cerebral diseases or symptoms, in particular stroke, access to a CT-scan should be easy. PMID- 11036448 TI - [Surgery for nearsightedness]. AB - Myopia can today be reduced or eliminated by refractive surgery. Excimer laser surgery of the cornea by surface sculpturing (photorefractive keratectomy) or intrastromal tissue removal (LASIK) are the most widely used techniques, although implantation of intra corneal ring segments for low myopia also appears promising. Treatment of high myopia (> 10 diopters) is still difficult although epikeratoplasty or phakic IOL implantation are present possibilities. The perfect surgery for myopia remains to be developed, but the existing techniques will without doubt be further optimised. In 10 years time, supra normal visual acuity may even be obtained when surgical, optical, and biological variables can be described and controlled in each individual undergoing refractive surgery. PMID- 11036449 TI - [Surgical treatment of otosclerosis with argon laser]. AB - OBJECTIVES: Argon laser stapedotomy is a new modality for the treatment of otosclerosis. The first results obtained in Denmark are reported. MATERIALS AND METHODS: A retrospective follow-up study was undertaken to evaluate the results of argon laser stapedotomy which was introduced in 1991 at the department of Otolaryngology, Rigshospitalet, Copenhagen. The results are based on 85 consecutive primary stapedotomy operations with argon laser in 76 patients during the period January 1, 1991 to June 30, 1996. Mean follow-up was 16.6 months (6 60). RESULTS: The postoperative air-bone gap was closed within 10 dB or less in 83% of the operations and in 20 dB or less in 98%. The mean speech reception threshold (SRT) was 48 dB preoperatively. All patients had an improved SRT after the operation, with a mean value of 23 dB. At 4000 Hz and 8000 Hz, 89% and 65% of the patients had maintained or improved their hearing. No patient developed profound sensorineural hearing loss or anacusis after the operation. Only one patient developed symptoms of a temporary perilymphatic fistula. CONCLUSION: Argon laser stapedotomy is a safe technique. Its complications are fewer and not as serious as conventional methods. In addition, the achieved hearing results are better, and thus it is a good example of "minimal invasive high success surgery". PMID- 11036450 TI - [Results and experiences with 55 cochlear implantations]. AB - INTRODUCTION: The etiology of deafness in deaf people is defects in the cochlea hair cells. Cochlear implant treatment gives deaf born and patients with acquired deafness the possibility to reestablish or obtain hearing by electric stimulation of the cochlear nerve. In this paper the results of treatment of 35 adults and 20 children are reported. METHOD: The candidates for cochlear implant treatment are extensively investigated before the decision is made to operate. The operative treatment takes place under general anaesthesia and the operative technique is outlined. In the treatment of the deaf, different types of implants and stimulation strategies have been used. The effect of treatment is considered by hearing test and evaluation of speech. RESULTS: Nine of the adults obtained a hearing quality that made using a telephone possible. Almost all adults can by the combination of cochlear implant and lip reading perform a normal conversation. The results in the deaf born children are obtained slowly as the deaf born have to develop hearing and speech from zero. All the treated children have improved possibilities of communication. DISCUSSION: Cochlear implant treatment of deaf people now seems well established both internationally and nationally in Denmark. Cochlear implant equipment has improved considerably throughout the past years and that it today is possible to establish hearing in deaf born or patients with acquired deafness can be considered as one of the greatest developments in otology, maybe the greatest. PMID- 11036451 TI - [Penicillin treatment of acute maxillary sinusitis in adults. A randomized, double-blind, placebo-controlled trial from general practice]. AB - INTRODUCTION: The aim of the study was to compare the effectiveness of penicillin V with placebo in the treatment of adult patients with acute maxillary sinusitis (in general practice). MATERIAL AND METHODS: The study was designed as a randomised, double-blind, placebo-controlled trial in 26 Danish general practices. The participants were 133 adult patients with acute maxillary sinusitis clinically diagnosed on maxillary pain and raised values of either C reactive protein (CRP) or the erythrocyte sedimentation (ESR) rate. Main outcome measures were pain score and illness score, and CRP and ESR values after initiation of treatment. RESULTS: Penicillin V led to a better recovery than placebo. The difference in pain reduction was statistically significant three days after initiation of treatment, whereas no significant difference was found in the reduction in the sense of illness. At the end of the study, significantly more patients in the penicillin group were completely free of pain than in the placebo group. This difference was found only in patients with an initial pain score of more than three. The cure rate was 71% in the penicillin group and 37% in the placebo group. Significantly more patients treated with penicillin achieved normal CRP values than those receiving placebo, respectively 88% and 75%. CONCLUSION: Penicillin V is more effective than placebo in the treatment of acute maxillary sinusitis in adults in general practice, but only in patients with pronounced pain. PMID- 11036452 TI - [Surgical results of submandibular gland excision]. AB - INTRODUCTION: Extirpation of the submandibular gland is the standard treatment for benign disease in cases where conservative treatment fails. This study attempts a follow-up focusing on: the effectiveness of the treatment, the number of complications and the long-term complaints related to this surgical procedure. MATERIAL AND METHODS: This study included 110 patients who, due to benign disease, underwent submandibular gland excision at our hospital during 1989-1995. All the patients had their charts reviewed and were invited for a follow-up consultation, with an examination and an interview about complaints arising after the procedure. RESULTS: Average follow-up time was 7.2 years (range 3.3 to 9.4 years) and 72 patients (65.5%) reported back for follow-up. Sixty-eight of the examined patients (92%) reported relief from their preoperative symptoms. Forty three patients (42%) had complications in connection with their surgery and fifteen patients (15%) needed a reoperation. The most frequent postoperative complaints were dryness of the mouth and reduced function of the marginal branch of the facial nerve. DISCUSSION: Most of the patients achieved relief of symptoms after surgery. There were a high number of complications, but most of these were of minor severity and temporary. The frequency of palsy in the submandibular branch decreased with time after operation while the frequency of oral dryness increased with time. Excision of the submandibular gland is an effective low-risk treatment for benign disease in the submandibular gland. PMID- 11036453 TI - [Surgical treatment of severe snoring. Long-term effects]. AB - AIM: The long-term results of uvulopalapharyngoplasty (UPPP) reported by patients who had been operated for severe snoring but without clinically significant obstructive sleep apnoea were explored. METHODS: Median three years (one to eight years) after the operation follow-up results were obtained from 80 of 85 patients (94%), using a questionnaire. RESULTS: In nine patients (11%) the operation had been without any effect. Eleven patients (14%) indicated that they were completely cured of snoring. In sixty patients (75%) snoring was reduced. In about half of these the initial effect had been more pronounced than the present. About 60% of the patients felt better during daytime, were less tired or sleepy than before the operation. Slight or temporary side effects were present in 30% of the patients. Eighty percent of the patients would recommend the operation to others. CONCLUSION: This study confirms that UPPP is effective in relieving the symptoms in non-apnoeic snorers, without significant side effects. A certain reduction of the positive effect occurs during the first two years in about half of the patients. Nevertheless most patients are satisfied with the operation and would recommend it to others. PMID- 11036454 TI - [Surgical treatment of hypernasality in Denmark]. AB - OBJECTIVE: A palatopharyngoplasty is a widely used operation performed to diminish hypernasality. The purpose of this study was to assess the results of the palatopharyngoplasties and to estimate if the type of cleft or other structural defect influenced these results. MATERIAL AND METHOD: This is a retrospective study of the patient records from 1/1-1991 to 1/11-1996 comprising 297 operations on which 284 records were available. The results of the operations were evaluated by speech pathologists who have rated the nasality before and after the palatopharyngoplasties. RESULTS: Following operation 90.1% of the patients obtained normal resonance or reduced hypernasality. The type of cleft or other structural defect did not influence the results. DISCUSSION: Palatopharyngoplasty is an effective operative procedure in the treatment of hypernasality, regardless of the type of cleft or structural defect. To further optimize the treatment a higher level of standardization is recommended. PMID- 11036455 TI - [Verbal autopsy: a method for epidemiologic studies in developing countries. Experiences from apoplexy cases]. PMID- 11036456 TI - [The mischievous eponyms]. PMID- 11036457 TI - [Loved child has many names]. PMID- 11036458 TI - [Prolonged pregnancy]. PMID- 11036459 TI - [The struggle for the existence and divergence of traits. 1940]. PMID- 11036460 TI - [The significance of adaptive modifications in evolution. 1940]. PMID- 11036461 TI - [A phenogenetic analysis of discreteness. 1941]. PMID- 11036462 TI - [The quantitative characteristics of animal adaptivity and the types of population dynamics in higher vertebrates. 1942]. PMID- 11036463 TI - [The effect of the nature of their dispersal on the course of insect ontogeny. 1945]. PMID- 11036464 TI - [The geographical variability in the population count dynamics and evolution. 1945]. PMID- 11036465 TI - [The diversity and unity of vital phenomena and quantitative methods in biology. 1981]. PMID- 11036466 TI - Primary yolk sac tumor of the rectum. AB - Extragonadal germ cell tumors are well recognized in men but have rarely been reported in women. Reports have primarily focused on the pediatric population and have suggested a poor prognosis for extragonadal yolk sac tumors. A 23-year-old woman with a yolk sac tumor arising in the rectum is described. A review of the English-language literature (MEDLINE 1966-1998) regarding extragonadal germ cell tumors in females is provided. Treatment with four courses of cisplatin, etoposide, and bleomycin was followed by surgical resection of the involved area. No residual tumor was identified. She remains disease free 3.5 years later. Previous reports are limited by the small number of patients, focus on the pediatric population, and treatment before the availability of cisplatin. Extragonadal germ cell tumors in women are extremely rare but can be successfully treated with aggressive chemotherapy and surgery similar to testis cancer. PMID- 11036467 TI - Survival of patients who had salvage castration after failure on bicalutamide monotherapy for stage (D2) prostate cancer. AB - Patients with hormone-naive stage D2 prostate cancer often benefit from castration. This treatment, however, frequently produces many unacceptable physical and psychological side effects, especially in younger and sexually active patients. Bicalutamide is an oral antiandrogen with excellent tolerance and preservation of sexual function. Three institutions participated in phase II and III trials of bicalutamide monotherapy (50 mg daily) as primary therapy in hormone-naive patients with stage D2 prostate cancer. Upon bicalutamide failure, all patients underwent castration and were followed until death. Fifty-four patients received bicalutamide 50 mg orally once a day. One patient (2%) had complete response, 9 patients (17%) had partial response, and 27 patients (50%) had stable disease. Seventeen patients (31%) had progressive disease. The median time to bicalutamide failure was 47.4 weeks, 70.5 weeks for the responders vs. 25.4 weeks for the nonresponders (p < 0.001). The median survival time after the sequential use of bicalutamide and castration was 119.2 weeks for all 54 patients, 162.0 weeks for the responders, and 73.5 weeks for nonresponders (p < 0.0001). The median survival time after initiation of castration was 71.1 weeks for all 54 patients, 91.4 weeks for bicalutamide responders, and 42.1 weeks for nonresponders (p < 0.01). In hormone-naive patients with stage D2 prostate cancer, sequential treatment with bicalutamide monotherapy followed by castration upon failure may produce survival time within the range reported for initial treatment with castration. Thus, considering the favorable quality of life profile of bicalutamide, further studies are needed to define the role of sequential hormonal therapy in younger sexually active patients. PMID- 11036468 TI - P-glycoprotein expression and multidrug resistance in cutaneous T-cell lymphoma. AB - Advanced-stage cutaneous T-cell lymphoma (CTCL) is generally resistant to standard chemotherapy. Because P-glycoprotein (P-gp) has been detected in other types of resistant solid tumors, leukemias, and lymphomas, we analyzed P-gp expression in CTCL. Twenty-seven patients with CTCL and circulating Sezary cells in the peripheral blood as observed on a peripheral smear treated at the Yale Photopheresis Center between 1987 and 1993 were identified. Twenty-five of these patients had skin biopsies evaluated for expression of P-gp using JSB-1 (Accurate Chemical), MRK-16 (gift of T. Tsuruo), and UIC-2 (gift of E. Metchner). P-gp expression was considered present if immunoreactivity was noted with two of the three antibodies. Eighteen of 25 patients (72%) evaluated exhibited expression. The patients were treated with various combinations of drugs consisting of topical and systemic steroids electron beam therapy, psoralens in combination with UV light A (PUVA), systemic chemotherapy, and photopheresis before testing the tissue for P-gp expression. Treatment with systemic chemotherapy in P-gp positive patients produced responses in 3 and no responses in 11 patients (4 were lost to follow-up). Seven patients did not express P-gp: One patient responded to treatment, five did not respond, and one patient was lost to follow-up. These results demonstrate that P-gp is frequently expressed in CTCL. P-gp expression in our study was not a useful predictor of drug resistance. PMID- 11036469 TI - Multidrug resistance phenotype and paclitaxel (Taxol) sensitivity in human renal carcinoma cell lines of different histologic types. AB - We compared the effects of paclitaxel (Taxol) in human renal cell carcinoma (RCC) of different histologic types. The growth inhibitory effects of paclitaxel on 34 human RCC cell lines of strictly defined different histologic types were determined by 3-[4,5-dimethylthiazolyl]-2,5-diphenyltetrazoliumbromide (MTT) assays. Paclitaxel-induced morphologic alterations were visualized by light and immunofluorescence and by transmission electron microscopy. The expression and function of P-glycoprotein and multidrug resistance-associated protein (MRP) were defined by reverse transcriptase polymerase chain reaction and fluorescence activated cell sorting (FACS) analysis, respectively. Modulation of P glycoprotein function was performed by verapamil or Cremophor EL. A significant (p < 0.05) dose-dependent paclitaxel-induced growth inhibition could be demonstrated in all cell lines, with the effects of paclitaxel dissolved in Cremophor EL/ethanol (= Taxol) exceeding the effects of paclitaxel dissolved in dimethyl sulfoxide. The extent of response markedly varied between the different cell lines, although chromophilic RCCs exhibited a more pronounced response to Taxol (IC50: 0.03-0.38 microM) than clear cell RCCs (IC50: 0.01-36.69 microM). Exposure to paclitaxel/Taxol induced an increase of microtubule bundles in the clear cell and the chromophobe RCCs but not in the chromophilic RCCs. The expression of the MRP was low in RCC cell lines and was not found to be related to paclitaxel/Taxol sensitivity. In contrast, the expression level of P glycoprotein was much more pronounced and showed a positive correlation (p < 0.05) with the response to paclitaxel. Reversal of P-glycoprotein function by verapamil or Cremophor EL enhanced the growth inhibitory effects of paclitaxel and further supported the role of P-glycoprotein for paclitaxel sensitivity of human RCCs. Paclitaxel/Taxol effectively inhibits proliferation of human RCCs in vitro, irrespective of their histologic types. Moreover, expression and function of P-glycoprotein markedly contribute to paclitaxel responsiveness, although other as yet undefined drug resistance mechanisms are effective in human RCCs as well. PMID- 11036470 TI - Evaluation of drug delivery and survival impact of dose-intense relative to conventional-dose methotrexate, vinblastine, doxorubicin, and cisplatin chemotherapy in urothelial cancer. AB - The efficacy of dose-intense methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC) chemotherapy relative to conventional-dose M-VAC in patients with advanced transitional cell carcinoma is unknown. The outcomes of 33 patients on two successive protocols using dose-intense M-VAC with granulocyte colony stimulating factor (G-CSF) support were compared with those of 129 patients treated with conventional-dose M-VAC to assess for an impact of dose-intense therapy on long-term survival. The mean relative dose intensity of chemotherapy delivered to the dose-intense cohort was 55% higher than that delivered to the conventional-dose cohort (p = 0.0001). However, no significant differences were observed with regard to response proportion (72% vs. 76%), median survival (13.3 vs. 16.7 months, p = 0.31), or 5-year survival (16% vs. 15%). Growth factor support enabled a statistically significant increase in the delivered dose intensity of M-VAC chemotherapy, but no survival advantage relative to conventional-dose M-VAC was observed. PMID- 11036471 TI - Antisense approaches for the treatment of cancer. PMID- 11036472 TI - Diagnostic investigations in cancer pain. AB - Defining the specific cause of pain in patients with cancer often has substantial therapeutic and prognostic implications. This process often requires the use of specific diagnostic investigations. Here we critically review the diagnostic investigations used in the evaluation of common pain problems in patients with cancer. Familiarity with this information facilitates the development of efficient and rational diagnostic strategies. PMID- 11036473 TI - A medical hypothesis: phosphorus balance and prostate cancer. AB - Over the last three decades the mortality rate for prostatic carcinoma has steadily increased. Carcinoma of prostate (CaP), the most common malignancy in men, is also the second most common cause of cancer deaths in men. However, few epidemiologic studies have been done, and there are scant clues to the etiology/pathogenesis of CaP. As treatment failures for advanced carcinoma continue to frustrate clinicians, more emphasis has recently been focused on strategies to prevent invasive CaP. Prostatic hyperplasia is a universal phenomenon in aging men. Mechanism and signals causing this growth are not understood. Thus, prostatic diseases affect men over the age of 45 and increase in frequency with age so that by the eighth decade more than 90% of men have benign prostatic hyperplasia, of which some progress to CaP. Data from several studies support that higher levels of active metabolite of vitamin D, 1,25-(OH)2 D, reduce the risk of prostatic hyperplasia and CaP. Men with high serum levels of 1,25-(OH)2-D have a reduced risk of poorly differentiated and clinically advanced CaP. Receptor for vitamin D has been reported in both normal and cancer prostate cells. 1,25-(OH)2-D inhibits proliferation and induces differentiation of normal and neoplastic cells. Hypercalcemic activity of 1,25-(OH)2-D or its analogues, however, thwart their use for therapy in humans. 1,25-(OH)2-D also has an established role in phosphorus homeostasis. Low dietary intake of phosphorus leads to an increase in serum concentration of 1,25-(OH)2-D. In addition, dietary fructose reduces plasma phosphate levels by 30 to 50% for more than 3 hr due to a rapid shift of phosphate from extracellular to intracellular compartment. Fruit intake has been shown to be associated with reduced risk of CaP, particularly the advanced type. Put together, these observations support that dietary determinants of hypophosphatemia, leading to increased plasma levels of 1,25-(OH)2-D, could reduce the risk of aging men to develop prostatic diseases, both benign prostatic hyperplasia and CaP. PMID- 11036475 TI - Tamoxifen for treatment of premenopausal women with breast cancer. PMID- 11036474 TI - The FAMMM syndrome: epidemiology and surveillance strategies. AB - Research into the epidemiology of the melanoma-prone FAMMM syndrome, molecular genetics of the occurrences of melanoma, the photobiology of DNA damage/repair, diagnostic epiluminescence, microscopic/imaging techniques, and a new concept of photoprotection have altered melanoma strategies in surveillance and prevention. Molecular genetic research has implicated the importance of hereditary aspects of melanoma and associated malignancies. High-risk pedigrees can be identified through an informatic analysis of the occurrence patterns of melanoma and systemic cancers in kindreds. All ultraviolet radiation results in cutaneous DNA damage and in high-risk individuals may cause melanoma. We may reverse the epidemic trend in melanoma occurrences in these high-risk pedigrees if we are willing to change our cultural approach to sunlight exposure with restrictive sunlight behavior, wearing of ultraviolet protective clothes, the use of broad spectrum ultraviolet protection from nightly topical dihydroxyacetone coupled with daytime UVB sunscreens, and periodic surveillance. PMID- 11036476 TI - Should tamoxifen be used to treat premenopausal women with breast cancer? PMID- 11036477 TI - Reducing toxicity of hormonal therapy for prostate cancer. PMID- 11036478 TI - Human experimentation in Judaism. AB - The supreme value of human life, the sanctity of human life, and the obligation of patients to accept standard efficacious medical therapy are axiomatic Judaic principles. If a seriously and/or terminally ill patient has already received all standard treatments and is asked to consider experimental therapy that may prolong his or her life or may hasten death, he or she is allowed but not required to accept the treatment. Healthy people may volunteer to participate in research studies that involve little or no risk. PMID- 11036479 TI - [DMW (Deutsche Medizinische Wochenschrift) Walter Siegenthaler Prize]. PMID- 11036480 TI - [DMW (Deutsche Medizinische Wochenschrift) Paul Boerner Prize]. PMID- 11036481 TI - [The whole width of cardiology]. PMID- 11036482 TI - [Brugada syndrome]. PMID- 11036483 TI - [Transthoracic echocardiography in endocarditis: a comparison of conventional and harmonic echocardiography]. AB - BACKGROUND AND OBJECTIVE: Compared with conventional echocardiography (CEC) harmonic imaging (HEC) provides better resolution of the endocardial line and valvular apparatus. This prospective study was undertaken to compare the value of harmonic and conventional transthoracic echocardiography in endocarditis and compare them with standard transoesophageal echocardiography (TOE) and operative findings. PATIENTS AND METHOD: Conventional and harmonic echocardiographic imaging was compared in 30 patients (aged 58 +/- 17 years; 19 males and 11 females) with endocarditis clinically judged to require surgical intervention. The results of both methods were then compared with those of standard TOE and the intraoperative findings. RESULTS: Intraoperatively 15 floating structures, 9 abscesses and 5 perforations were demonstrated. Transoesophageal echocardiography was better than the standard method in diagnosing floating structures and detecting abscesses (15 vs. 10 vegetations, p < 0.05; 7 vs. 5 abscesses, p = 0.05). TOE was better than either method in the diagnosis of abscess. CONCLUSION: Harmonic transthoracic echocardiography is better than the conventional mode in diagnosing complications of endocarditis. PMID- 11036484 TI - [Left ventricular diastolic function in normal pregnancy. A prospective study using M-mode echocardiography and Doppler echocardiography]. AB - BACKGROUND AND OBJECTIVE: During pregnancy many substantial changes occur in the cardiovascular system. Aim of this study was to examine how physiological preload alterations influence left ventricular haemodynamic parameters. PATIENTS AND METHODS: During the 9th, 24th and 33rd weeks of pregnancy and 8 weeks after childbirth 36 patients underwent echocardiographic studies. 36 young not pregnant women (25 +/- 7 years) served as controls. The following Doppler echocardiographic parameters were measured: peak early diastolic flow velocity (VE, ms); acceleration (AT; ms) and deceleration time (DT; ms) of flow velocity in early diastole; peak late diastolic flow velocity (VA; m/s) and isovolumetric relaxation time (IVRT; ms). In all women the left ventricular muscle mass index (LVMMI), fractional shorting (FS; %) and the ratio between septum and posterior ventricular wall were calculated. RESULTS: During pregnancy all women showed an elevation of the left ventricular muscle mass index (LVMMI: from 66 +/- 6 to 100 +/- 9 g/m2; p < 0.01) and a decrease of fractional shortening (FS: from 38 +/- 4 to 31 +/- 3%). All patients developed a relevant diastolic dysfunction: reduced early diastolic flow velocity (VE: from 0.89 +/- 0.11 to 0.83 +/- 0.19 m/s; P < 0.01), reduced E/A ratio (1.7 +/- 0.4 to 1.2 +/- 0.4; P < 0.01), prolonged IVRT (72 +/- 12 to 114 +/- 12 ms; P < 0.01) and deceleration time (DT: to 189 +/- 17 to 227 +/- 18 ms; P < 0.01). Eight weeks after childbirth all parameters of left ventricular systolic and diastolic functions were normal. CONCLUSION: Preload alterations during normal pregnancy lead to reversible physiological left ventricular hypertrophy. Furthermore, we found a short-time reduction of systolic function just before childbirth and a significant alteration of the left ventricular diastolic filling pattern (abnormal relaxation pattern). While left ventricular systolic function was normal in all patients one week after childbirth, left ventricular hypertrophy and left ventricular diastolic dysfunction persisted for nearly two months. PMID- 11036485 TI - [Recurrent syncope in a 34-year-old woman triathlete]. AB - HISTORY: Some weeks previously a 34-year old athlete, specializing in the triathlon, had 6 syncopes in one day. They had caused abrasions and contusions resulting from the falls. At another hospital paroxysmal atrial fibrillation had been diagnosed and treatment with disopyramide (2 x 200 mg) initiated, but she had about 15 further syncopes within 2 weeks. She was admitted for establishing their cause. INVESTIGATIONS: Initial ECGs and neurological examination failed to provide a diagnosis and she was discharged with an "event recorder". DIAGNOSIS, TREATMENT AND COURSE: Three weeks after discharge she had another syncope. The event recorder was activated by the patient's partner and revealed polymorphous ventricular tachycardia. She underwent extensive invasive cardiological tests, including a right ventricular biopsy, but no abnormality was demonstrated. However, a provocation test with ajmaline produced ST segment elevations in V1 and V2 typical of the syndrome previously described by the Brugadas (right bundle branch block, precordial ST elevations in V1-V3 and sudden cardiac death). A cardioverter-defibrillator was implanted. During the subsequent observation period of 2 month the ICD delivered one countershock, triggered by the onset of polymorphous ventricular tachycardia with syncope. CONCLUSION: In patients with serious ventricular arrhythmias but no diagnostic findings, including a normal resting ECG, a drug provocation test should be performed to exclude a Brugada syndrome. PMID- 11036486 TI - [New possibilities by means of electronic DMW (Deutsche Medizinische Wochenschrift) pages]. PMID- 11036487 TI - [Ergometry after acute myocardial infarct]. PMID- 11036488 TI - [Estrogen replacement therapy and coronary disease]. PMID- 11036489 TI - [Infection thesis as explanation of the higher incidence of myocardial infarction in males]. PMID- 11036490 TI - [Self-medication--economic, sociopharmacologic and toxicological aspects]. PMID- 11036491 TI - Mycobacterium leprae typing by genomic diversity and global distribution of genotypes. AB - The genetic diversity and related global distribution of 51 Mycobacterium leprae isolates were studied. Isolates were obtained from leprosy patients from 12 geographically distinct regions of the world and two were obtained from nonhuman sources. Polymerase chain reaction (PCR) followed by DNA sequencing was performed targeting the rpoT gene of M. leprae. Isolates were classified into two groups based on the number of tandem repeats composed of 6 base pairs in the rpoT gene. Isolates from Japan (except Okinawa) and Korea belonged to one group, while those from Southeast Asian countries, Brazil, Haiti and Okinawa in Japan belonged to a second genotype. M. leprae obtained from two nonhuman sources (an armadillo and a mangabey monkey) revealed the latter genotype. These results demonstrate the genetic diversity of M. leprae and the related genotype-specific distribution in the world. PMID- 11036492 TI - Trials of daily, long-term minocycline and rifampin or clarithromycin and rifampin in the treatment of borderline lepromatous and lepromatous leprosy. AB - Daily, long-term treatment with minocycline 100 mg and rifampin 600 mg was initiated in 24 previously untreated borderline lepromatous (BL) and lepromatous (LL) patients for a total of 646 patient-months, averaging 26.9 months per patient. The same regimen was started in 12 BL and LL patients having a bacteriologic relapse for a total of 379 patient-months, averaging 32.5 months per patient, and in 12 patients judged to be at high risk for relapse for a total of 354 patient-months, averaging 29.5 months per patient. Daily, long-term treatment with clarithromycin 500 mg and rifampin 600 mg was initiated in 8 previously untreated BL and LL patients for a total of 174 patient-months, averaging 21.8 months per patient. The results in these 56 patients were compared to those obtained in 34 previously untreated BL and LL patients who were treated concurrently receiving daily, long-term dapsone 100 mg and rifampin 600 mg. No evidence of dangerous drug reactions or bone marrow, kidney or liver toxicity was seen in any of these five patient groups. Drug intolerance in 10 of the 90 patients studied necessitated discontinuing the chosen regimen, 4 from rifampin, 3 from dapsone, 2 from minocycline and 1 of undetermined attribution. The use of either minocycline or clarithromycin in conjunction with rifampin appears to pose no great risk when used long term. PMID- 11036493 TI - Induction of lepromin positivity and immunoprophylaxis in household contacts of multibacillary leprosy patients: a pilot study with a candidate vaccine, Mycobacterium w. AB - We screened 487 household contacts of multibacillary (MB) patients for evidence of disease and their lepromin status. From the 444 results available, 302 (68.02%) were lepromin positive and 142 (31.98%) were lepromin negative on initial testing. The initial lepromin status as assessed in the group of 54 contacts having disease at the outset showed 24 out of 46 (52.2%) to be lepromin positive and 22 of 46 (47.8%) to be lepromin negative. In the same group, among 24 lepromin positives, 22 (91.7%) had paucibacillary (PB) and 2 (8.3%) had multibacillary (MB) disease; among the lepromin negatives, 12 (54.5%) had PB and 10 (45.5%) had MB disease. Out of 72 initially lepromin-negative contacts administered Mycobacterium w vaccine and followed up, the cumulative percentages show that 53 (73.6%) converted to positivity after a single dose, 10 (87.5%) after a second dose and 67 (93.1%) after the third dose. The incidence of new cases with leprosy was 8 out of 231 (3.46%) among lepromin-positive contacts and 5 out of 93 (5.38%) among lepromin-negative contacts administered Mycobacterium w vaccine. Among 231 lepromin-positive contacts, the new cases occurred in those with a 1+ and 2+ lepromin response only, and no case occurred among 51 contacts with a 3+ lepromin response. The incidence among lepromin-positive contacts in this study (3.46%) was similar to the observations in two other studies: 3.2% by Dharmendra, et al. and 6.9% by Chaudhary, et al. However, the incidence among lepromin-negative contacts administered Mycobacterium w vaccine was significantly lower than that observed among lepromin-negative contacts not administered any vaccination in the other two studies (14.1% by Dharmendra, et al. and 29.0% by Chaudhary, et al.). To conclude, although a study of small sample size, the preliminary evaluation indicates that administration of Mycobacterium w vaccine seems to have the potential to reduce the incidence of leprosy among household contacts of leprosy patients. More explicit results about the vaccine will be available from the ongoing field trials in Kanpur Dehat in the near future. PMID- 11036494 TI - Distinct patterns of microvasculature in the cutaneous lesions of leprosy. AB - This work is an investigation on the microvasculature of the cutaneous infiltrates of leprosy with the immunohistochemical staining of endothelial cells in cutaneous biopsies. Anti-Factor VIII-related antigen antibody (anti-FVIII-ra) and Ulex Europaeus-1 lectin (UEA-1) binding were utilized as endothelial cell markers. Thirty-nine patients grouped according to the Ridley-Jopling classification (14 borderline tuberculoid, 18 borderline lepromatous, 6 lepromatous, and 1 indeterminate leprosy) were selected for this study. Two microvascular architectural patterns could be clearly distinguished: lepromatous lesions presented a dense and tortuous mesh of microvessels among the Mycobacterium leprae-glutted macrophages; whereas the microvessels in the tuberculoid lesions were restricted to the periphery of the granulomas and were not seen among the central epithelioid cells. We were able to distinguish three basic morphological kinds of infiltrate distribution related to the microvessels: micronodules, cords and macronodules. Intensifications of the FVIII-ra immunoreactivity and UEA-1 binding capacity were observed in the endothelial cells of microvessels involved by the inflammatory infiltrate. A distinct cytokine expression profile at the leprosy poles and the role of mast cells in angiogenesis were speculated as factors contributing to these distinct patterns. Growth of the lesion and systemic dissemination of M. leprae in the bipolar spectrum of leprosy may hypothetically be influenced by the vascular-infiltrate relationship. The detection of angiogenesis in the cutaneous lesions of leprosy may bring about alternate and/or additional strategies for leprosy treatment. PMID- 11036495 TI - Epithelioid granuloma in the iris of a lepromatous leprosy patient; an unusual finding. AB - This case report depicts a case of histopathologically confirmed polar lepromatous (LL) leprosy with a bacterial index of 4+. He experienced recurrent episodes of erythema nodosum leprosum (ENL) in the first 5 years after diagnosis. Skin smears became negative after 6 years of dapsone monotherapy and have remained negative since that time. At 23 years after diagnosis, the patient had developed cataracts and underwent intracapsular cataract extractions with broad based iridectomies. In one of the iris specimens, histopathologic examination revealed a focal granuloma composed of epithelioid cells. Subsequently a lepromin skin test showed a positive Mitsuda reaction with a borderline tuberculoid histopathology. This clearly illustrates the immunological upgrading of a polar lepromatous patient, perceived first in the iris tissue. PMID- 11036496 TI - Experimental Mycobacterium leprae infection in BALB/c mice: effect of BCG administration on TNF-alpha production and granuloma development. AB - In the present study, the experimental model of Mycobacterium leprae infection in the foot pads of BALB/c mice was used to investigate the effects of BCG administration on tumor necrosis factor-alpha (TNF-alpha) production and granuloma development. It was observed that mice intravenously infected with BCG 7 months after M. leprae inoculation into the foot pads presented a more effective mycobacteria clearance, revealed by a significant reduction of BCG colony forming units in the spleen and by the reduction of acid-fast bacilli (AFB) in the foot pads. BCG infection at the peak of M. leprae infection also modulated the granulomatous response to M. leprae by converting mononuclear granulomas into an epithelioid-cell granuloma. Furthermore, lower TNF-alpha serum levels were detected in M. leprae-infected mice when compared to mice infected with M. leprae + BCG. An analysis of the TNF-alpha gene expression in the spleen by semiquantitative reverse transcription-polymerase chain reactions (RT-PCR) demonstrated that co-infection with BCG induced an earlier expression of TNF alpha mRNA than in M. leprae-infected mice. The numbers of TNF-alpha-positive cells and apoptotic cells were also enhanced in epithelioid versus non epithelioid granulomas. As a whole, the data suggest that co-infection of M. leprae-infected mice with BCG modulates TNF-alpha synthesis which, in turn, leads to induction of protective epithelioid granuloma formation in the foot pads and subsequent mycobacterial clearance. Macrophage differentiation into epithelioid cells, in association with the enhancement of TNF-alpha production at the granuloma site, may represent a triggering signal that induced apoptosis in these cells, leading to mycobacterial elimination. Moreover, the rate of apoptosis in epithelioid granulomas may well be related to the extent of immunopathologically mediated tissue damage. PMID- 11036497 TI - Histological assessment of dermal nerve damage occurring during multidrug therapy for leprosy. AB - This is a prospective histomorphological assessment of dermal innervation in biopsies taken before and after multidrug therapy (MDT) from 41 leprosy patients: 35 borderline tuberculoid (BT), 3 borderline lepromatous (BL), 3 lepromatous (LL). Biopsies of the same lesions taken before commencement (diagnostic therapy) and at the end of therapy (check biopsy) were compared. Hematoxylin and eosin, immunoperoxidase stain for S-100 protein, and the Holmes' silver impregnation method for nerve cells and fibers were used. Skin biopsies were classified as having detectable or undetectable nerves. Of 35 patients with BT leprosy, 17 had no detectable nerves in their diagnostic biopsies; in the check biopsies of 13 of these 17, dermal nerves remained undetectable, in 2 they were S-100 positive but were Holmes negative. Identifiable dermal nerves were present in diagnostic biopsies from 18 patients; in the check biopsies 5 of these 18 had no detectable nerves while in the remaining 13 nerve branches could be detected. The study provides histological documentation of complete damage to dermal innervation in 62.85% (22/35) of patients with BT leprosy, of which 14.28% (5/35) occurred during MDT. Of the patients with detectable dermal innervation at the onset of MDT, 27.7% (5/18) suffered continuing damage during MDT. PMID- 11036499 TI - Comments on leprosy at age 141. PMID- 11036498 TI - Survey of newly diagnosed leprosy patients in native and foreign residents of Japan. PMID- 11036501 TI - A study of the biology of M. lufu and prospects for using it in leprosy investigations. PMID- 11036500 TI - Multidrug therapy in geriatric patients. PMID- 11036502 TI - Disabilities in leprosy. PMID- 11036503 TI - Clinical and histopathological correlation in leprosy. PMID- 11036504 TI - Microcystins (cyanobacterial toxins) in drinking water enhance the growth of aberrant crypt foci in the mouse colon. AB - Microcystis aeruginosa produces toxic cyclic peptides called microcystins, potent hepatotoxins that have been implicated in tumor promotion in skin and liver. The model used in this investigation was the azoxymethane (AOM)-induced aberrant crypt focus (ACF) in the male C57Bl/6J mouse colon. Three intraperitoneal (i.p.) injections of 5 mg/kg AOM were administered at 7-d intervals to mice; 19 d after the last AOM injection, drinking water containing Microcystis extract was commenced and continued for a further 212 d. The content of microcystins in the drinking water was determined by mouse bioassay, high-performance liquid chromatography (HPLC), capillary eletrophoresis, and protein phosphatase inhibition. The doses employed were 0, 382, and 693 micrograms/kg bodyweight/d at the midpoint of the trial. Following postmortem examination blood cells, serum enzymes and organ pathology were investigated. A significant microcystin dose dependent increase in the area of aberrant crypt foci was observed. There was no marked increase in the number of crypts/colon. Two overt colonic tumors (approximately 30 mm3) were seen in microcystin-treated mice, and one microscopic colonic tumor in an AOM-alone-treated mouse. This investigation provides the first evidence for the stimulation of preneoplastic colon tumor growth by microcystin. PMID- 11036505 TI - Evaluation of oxidative stress in experimental colitis: effects of L-arginine nitric oxide pathway manipulation. AB - In this study it was of interest to evaluate the impact of nitric oxide (NO) modulation by administration of arginine/NAME, on oxidative stress in experimental colitis induced by 2,4,6-trinitrobenzenesulfonic acid. Arginine was used to increase NO levels while NAME lowered oxidant levels. Histopathological findings of colon revealed mucosal inflammation in all groups but significantly higher with arginine alone. The levels of NO and of thiobarbituric acid-reactive substances (TBARS, a marker of lipid peroxidation) were observed to be significantly higher in the arginine-administered group compared to glycine, and these levels were found to decrease on administration of NAME to both glycine- and L-arginine-administered groups. Glutathione peroxidase (GSH-Px) activity and glutathione (GSH) levels were significantly higher in arginine administered group compared to glycine. Significantly higher CuZn superoxide dismutase (CuZn-SOD) activity was observed in the L-arginine + L-NAME group compared to arginine. Data show that NO plays a role in oxidant damage found in experimental colitis and that the use of NAME may potentially inhibit injury. PMID- 11036506 TI - Ultraviolet radiation and reactive oxygen generation as inducers of keratinocyte apoptosis: protective role of tea polyphenols. AB - Ultraviolet A (UVA) radiation produces serious damage to skin, especially to dermis, but its damage to epidermis and responsible mechanisms are not fully understood. Studies were thus undertaken to investigate the effects of UVA or reactive oxygen species (ROS) on lipid peroxidation, cell cycle, and apoptosis in primary cultured rat keratinocytes and to determine the possible protective effects of tea polyphenols (TPP). UVA or ROS increased the release of plasma enzyme lactate dehydrogenase (LDH), and increased lipid peroxidation production (malondialdehyde, MDA), but decreased the activity of glutathione peroxidase (GSH Px), indicating that UVA or ROS were cytostatic and peroxidizing to keratinocytes. TPP stabilized and protected cell membranes from ROS or UVA by inhibiting the release of LDH, lowering MDA levels, and increasing GSH-Px activity. Flow cytometry (FCM) analysis revealed that UVA or ROS decreased the proliferative index (PI); hence the cell growth was blocked in the S/G2 phase, with an increase in the percentage of apoptosis in primary keratinocytes. TPP modified the UVA or ROS-induced changes in PI and apoptosis. TPP may be useful to protect keratinocytes from UVA irradiation. In summary, these data demonstrated that UVA damage to skin keratinocytes in vitro was similar to that for ROS and that TPP protects against UVA-induced cytotoxicity by inhibiting lipid peroxidation and apoptosis. PMID- 11036507 TI - In vitro penetration of soil-aged mercury through pig skin. AB - The dermal bioavailability of mercury "aged" in soil for 3 mo was compared to that of pure mercury (without soil) and to mercury in brief contact with soil (16 h). Studies were conducted in vitro with [203Hg]mercuric chloride on dermatomed male pig skin by flow-through diffusion cell methodology. Less than 0.5% of the initial mercury dose penetrated through skin into receptor fluid after each treatment. The majority of pure mercury became covalently bound to skin. However, a short contact time with either an Atsion (sandy) or Keyport (clay) soil significantly decreased the total penetration of mercury (sum of receptor fluid and skin) by 40%. After aging, a 95% reduction in total penetration was observed for the compound relative to chemical without soil. Both soils bind mercury more strongly with time, as evidenced by larger quantities of radioactivity in soil and smaller amounts in skin decontaminate after aging than in soil for 16 h. Decreased mercury bioavailability with aging indicates lower health risk and reduced need for soil cleanup. PMID- 11036508 TI - Stimulation of MCF-7 cell proliferation by low concentrations of Chinese domestic polychlorinated biphenyls. AB - The simple and sensitive in vitro MCF-7 human breast cancer cell proliferation assay was used to examine the proliferation abilities of two Chinese commercial polychlorinated biphenyl (PCB) mixtures made in the 1960s. Chinese PCB3 and Chinese PCB5 were compared with 17 beta-estradiol (E2). All of the positive activities of these types of Chinese PCBs were significantly different compared to controls with respect to MCF-7 cell doubling time. At lower levels of 7.8 pg/ml and 182 pg/ml, the Chinese PCB3 showed 94% and 86% of relative proliferation effects compared to 17 beta-estradiol, respectively. Chinese PCB5, also showed higher cell proliferation activity at lower level of 8.3 pg/ml, with relative proliferation effect as high as 107% in comparison to 17 beta-estradiol. Thus, both PCBs seem to be different from corresponding Aroclor mixtures. However, Chinese PCBs did not express cell proliferation effects at higher levels of 9.1 ng/ml for Chinese PCB3 and 166 pg/ml and 8.3 ng/ml for Chinese PCB3. This may be due to cytotoxicity and/or antiestrogenic compounds in the mixtures. PMID- 11036509 TI - Physiologically based modeling of the maximal effect of metabolic interactions on the kinetics of components of complex chemical mixtures. AB - The objective of this study was to predict and validate the theoretically possible, maximal impact of metabolic interactions on the blood concentration profile of each component in mixtures of volatile organic chemicals (VOCs) [dichloromethane (DCM), benzene (BEN), trichloroethylene (TCE), toluene (TOL), tetrachloroethylene (PER), ethylbenzene (EBZ), styrene (STY), as well as para, ortho-, and meta-xylene (p-XYL, o-XYL, m-XYL)] in the rat. The methodology consisted of: (1) obtaining the validated, physiologically based toxicokinetic (PBTK) model for each of the mixture components from the literature, (2) substituting the Michaelis-Menten description of metabolism with an equation based on the hepatic extraction ratio (E) for simulating the maximal impact of metabolic interactions (i.e., by setting E to 0 or 1 for simulating maximal inhibition or induction, respectively), and (3) validating the PBTK model simulations by comparing the predicted boundaries of venous blood concentrations with the experimental data obtained following exposure to various mixtures of VOCs. All experimental venous blood concentration data for 9 of the 10 chemicals investigated in the present study (PER excepted) fell within the boundaries of the maximal impact of metabolic inhibition and induction predicted by the PBTK model. The modeling approach validated in this study represents a potentially useful tool for screening/identifying the chemicals for which metabolic interactions are likely to be important in the context of mixed exposures and mixture risk assessment. PMID- 11036510 TI - [Hypernatremia in the aged: clinical characteristics]. AB - OBJECTIVE: Hypernatremia in young adults is a rare condition, sometimes occurring after gastrointestinal diseases, osmotic diuresis or diabetes insipidus. The clinical characteristics and prognosis of hypernatremia in geriatric patients would be different. Each case must be examined separately. PATIENTS AND METHODS: We conducted a prospective uncontrolled study among patients over 70 with sodium levels equal to or greater than 150 mmol/l (at admission or during the hospitalization). RESULTS: Hypernatremia was found in 77 patients. Mean age was 84.1 +/- 6.7 years. The mean peak serum sodium level was 157.3 +/- 7.4 mmol (two thirds ranged from 150 to 160). Hypernatremia onset was produced by a wide range of symptoms: 48% had a febrile illness, 470% were on diuretics, 15% had had a stroke, but only 9% were suffering from diarrhea or vomiting. Most of the patients had disabling chronic illnesses (62% loss of two or more ADL of Katz), severe dementia (84% level = 6 on the Reisberg global deterioration scale). In spite of fluid replacement and follow-up treatment, outcome was poor: 62% of the patients died within three months. CONCLUSION: Besides digestive and renal losses, hypernatremia in elderly patients is increased by fluid supply disorders consecutive to hypodipsia contracted at the same time as disability and dementia. Hypodipsia is a sign of poor prognosis. Prevention must be undertaken early, emphasizing the importance of identifying dehydration early among elderly people with chronic disabling illnesses. PMID- 11036511 TI - [Absolute cardiovascular risk in diabetes]. AB - OBJECTIVE: Diabetes is a main cause of cardiovascular morbidity and mortality. The aim of this study was to evaluate the influence of diabetes on cardiovascular risk using prediction equations. PATIENTS AND METHODS: The cardiovascular risk of 356 diabetics was calculated from the prediction equations established by the Framingham, Ducimetiere, Laurier-Chau and PROCAM studies, with and without the factor "diabetes". Calculations were made according to the conditions of each equation then by applying formulae to the whole population. DISCUSSION: Diabetes increased the cardiovascular risk at 4 years from 0.7% with the Ducimetiere equation to 3.6% to 8% with the Framingham equation. Formulae increased the cardiovascular risk when they were applied to the whole population. Models of prediction did not take into account however duration of diabetes and balance, parameters which are closely linked to cardiovascular risks. CONCLUSION: Absolute cardiovascular risk, whose practical interest has often been underlined, therefore has limitations in diabetes. PMID- 11036512 TI - [Severe chronic renal failure subsequent to acute pyelonephritis in alcoholic patients]. AB - BACKGROUND: Chronic alcoholism and malnutrition are uncommon causes of complicated acute pyelonephritis (APN). CASE REPORTS: Since 1997, we have seen 5 patients with chronic alcoholism (3 women and 2 men, mean age 53.4 +/- 13 years) without cirrhosis, diabetes or renal failure who developed severe APN in a state of malnutrition (albumin 22 +/- 3 g/l, total cholesterol 0.86 +/- 0.2 g/l). Diagnosis was made 14.6 +/- 9 days after onset of atypical symptoms which the patients neglected. There was a major bacterial inoculum: Escherichia coli 10(6.2 +/- 2) (3 multisusceptible and 2 amoxicillin-resistant strains); positive blood cultures in 3 cases. The imaging study showed bilateral diffuse lesions with focal swelling and kidney enlargement, without obstacle, abscess, or papillary necrosis. All patients had severe acute renal failure (maximum serum creatinine: 582 +/- 210 mumol/l; 3 patients underwent dialysis). Mean duration of antibiotic therapy was 40 +/- 7 days (i.v.: 22 +/- 3 d). Renal scarring occurred since creatinine clearance was 33 +/- 22 ml/min 2 months after the initial episode. One patient progressed to end-stage renal failure. CONCLUSION: In malnourished alcoholic patients, APN may be unusually severe due to late diagnosis leading to the risk of irreversible renal damage and severe chronic renal failure. PMID- 11036513 TI - [Severe thrombopenia during digitoxin therapy]. PMID- 11036514 TI - [Diabetes in trisomy X: an unknown association?]. PMID- 11036515 TI - [Spondylodiscitis caused by Enterobacter aerogenes]. PMID- 11036516 TI - [The ELITE 2 study]. PMID- 11036517 TI - [Drug watch in the hospital]. PMID- 11036518 TI - [Management of patients with malignant mesothelioma of the pleura]. PMID- 11036519 TI - [Alveolar echinococcosis: a dreadful orphan disease]. AB - A WIDESPREAD DISEASE: Significant progress in screening for alveolar echinococcosis has reduced the number of new cases observed in Europe. Health education and serodetection campaigns have allowed earlier diagnosis and more effective treatment. The disease cannot however be totally eradicated due to the widespread wild reservoirs, sometimes even in the center of large cities. THERAPEUTICS: Early diagnosed and treatment can inhibit the inevitable progression observed after clinical manifestations appear. Drugs can block disease progression and surgical excision can be most effective. Inversely, the hopes raised by liver transplantation in patients with advanced stage disease have not been fulfilled due to the more or less late-onset metastasis favored by immunosuppressive treatments. PERSPECTIVES: There has been considerable progress in our knowledge of this parasite disease, particularly in improved diagnostic techniques. They have also demonstrated that humans are poor hosts for the parasite which is often spontaneously ejected. We are beginning to better understand the mechanisms of this spontaneous cure. Practical consequences would be a definition of receptive patient profiles or "vaccine" or immunotherapeutic procedures. PMID- 11036520 TI - [Impact of Chlamydia pneumoniae infections on asthma]. AB - VIRUSES AND BACTERIA: The fact that the airways are exposed to a large number of infectious agents could explain the frequency of respiratory infections and their causal effect in bronchial inflammation. Viruses are most often the causal agent, but the frequency of bacterial infections make them potential candidates in certain respiratory diseases. Chlamydia are particularly important due to their capacity to provoke immune dysfunction and chronic inflammation. EFFECT ON ASTHMA: It is not surprising to find biological evidence of Chlamydia pneumoniae infection in a large number of subjects who experience major degradation of their asthma because asthmatic subjects are particularly susceptible to respiratory infections and Chlamydia pneumoniae is a frequent cause of such infections. PATHOGENIC EFFECT: Finding Chlamydia pneumoniae as the causal agent in asthma is however much more surprising, with a much different consequence. There are however many epidemiological and clinical findings and case observations (Chlamydia pneumoniae asthma associations, prolonged favorable course in certain obstructive bronchial diseases after a short antibiotic regimen) as well as provocative pathophysiological data favoring this particular form of "infectious asthma". FURTHER INFORMATION: Large-scale studies with rigorous methodology remain to be performed. The would be needed to determine the exact relationship between Chlamydia pneumoniae infections and certain types of asthma, particularly when wheezing occurs after a respiratory infection and when chronic obstruction develops. The could also determine the role of anti-Chlamydia pneumoniae antibiotics in case of obstructive respiratory failure and also determine their efficacy on long-term outcome. PMID- 11036521 TI - [Standards, options and guidelines for management of patients with malignant mesothelioma of the pleura]. PMID- 11036522 TI - [Tophaceous gout]. PMID- 11036523 TI - [Percentage of asthmatic patients with acute bronchitis and Chlamydia pneumoniae seropositivity]. PMID- 11036524 TI - [Protein import into mitochondria]. AB - This review is focused on the import of processable precursor proteins into the mitochondrial matrix; the import of carrier proteins into the inner mitochondrial membrane is also briefly discussed. Post- and cotranslational theories of the import, specific features of the presequence structures, and effects of some cytosolic factors on the import of precursor proteins are reviewed. The data on the structure of the protein translocases of the outer (TOM complex) and the inner (TIM complex) membranes of mitochondria and the current models of the precursor protein import by these translocases are also summarized. PMID- 11036525 TI - [MALD-MS in the quantitative analysis of peptides and proteins]. AB - A modified method of isotope dilution was applied to the quantitative determination of peptides and proteins by MALDI MS at subpicomolar level. The essence of the method consists in the quantitative analysis of the enzymic hydrolysis products rather than the starting compounds. This allows the measurements to be performed at a higher resolution and makes the method independent of the molecular mass of oligopeptides and proteins examined. Fragments obtained by hydrolysis of the same oligopeptide or protein in a known concentration by the same enzyme and labeled with the stable 18O isotope are used as internal standards. The label is introduced by carrying out the hydrolysis in H(2)18O, and the oligopeptide concentration is calculated from the isotope distribution between the labeled and unlabeled hydrolysis products in the mass spectrum. This method was tested in the determination of concentrations of the angiotensinogen (1-14) fragment (oligopeptide), extracellular RNAase from Bacillus amyloliquefaciens (protein) and its protein inhibitor, barstar M. Usefulness of this method in kinetic studies was also demonstrated. PMID- 11036526 TI - [Ribonuclease from Bacillus thuringiensis var. subtoxicus. Gene structure and biosynthesis regulation]. AB - The gene for extracellular guanyl-specific ribonuclease of Bacillus thuringiensis var. subtoxicus (RNase Bth), a close homologue of the B. intermedius RNase (binase), was completely sequenced. Analysis of nucleotide sequences in the regions adjoining RNase genes revealed an identical organization of the chromosomal loci of RNase Bth and binase. Growth characteristics of the Bacillus thuringiensis var. subtoxicus strain and its synthesis of RNase were studied. It was shown that the exogenous inorganic phosphate inhibits the biosynthesis of RNase. At the same time, actinomycin D in low doses stimulates the enzyme synthesis. Comparative analysis of the influence of inorganic phosphate and actinomycin D on the biosynthesis of RNAse Bth and binase suggests a possibility of coincidence of regulatory pathways of synthesis of these enzymes. PMID- 11036527 TI - [Chemical ribonucleases. 2. Design and hydrolytic properties RNase mimetics based on diazabicyclo[2.2.2]octane with various positive charges]. AB - A procedure was proposed allowing one to synthesize RNA mimics on the basis of conjugates of diazabicyclo[2.2.2]octane with imidazole bearing a varying number of positive charges (nDm series, where n is the number of positive charges at neutral pH, m is the code of an imidazole-containing fragment of the catalytic domain: 1, histamine; 2, histidine methyl ester). The hydrolytic activity of six compounds of this series was studied under physiological conditions using in vitro transcript of human mitochondrial tRNA(Lys) as a substrate. It was shown that the rate of RNA hydrolysis with nDm conjugates rises with an increase in the number of positive charges: an approximately 30-fold acceleration of hydrolysis was observed with an increase in the total charge of the construct from +2 to +4. PMID- 11036528 TI - [Isolation and structural study of polysaccharides from campion Silene vulgaris]. AB - Silenan SV, a pectic polysaccharide, was isolated from the aerial part of Silene vulgaris (Moench) Garke (Oberna behen (L.) Ikonn.), widespread through the European North of Russia. The polysaccharide was found to contain residues of galacturonic acid (63%), arabinose, galactose, and rhamnose as the main constituents. The results of a partial acidic hydrolysis, pectinase digestion, and NMR studies of silenan SV indicated that its molecule contains a linear alpha 1,4-D-galacturonan backbone and ramified regions. The core of the ramified regions is composed of residues of alpha-1,4-D-galacturonic acid along with 2 substituted alpha-rhamnopyranose residues. The NMR data showed that the silenan SV side chains are composed of the blocks built from the terminal alpha-1,5 linked arabinofuranose and beta-1,4-linked galactopyranose residues; these most likely are the side chains of rhamnogalacturonan, characteristic of other pectic polysaccharides. The nonreducing ends of these side chains contain alpha arabinofuranose residues. PMID- 11036529 TI - [Synthesis and properties of fluorescent labeled detergents analogues of glycocholic acid]. AB - For studying membrane processes with participation of detergents, fluorescent analogues of glycocholic acid containing p-hydroxybenzyl, 7-nitrobenz-2-oxa-1,3 diazol-4-yl, or fluorescein-5-thiocarbamoyl fluorophore in the glycyl moiety attached to glycocholic acid were synthesized. The fluorophores are in the probes near their carboxyl groups and, in membrane systems, should therefore be situated on the interface and be sensitive to phase transitions. The critical micelle concentrations were determined for the analogues and found to be close to those of cholate and glycocholate in the case of the first two compounds. We presume that the behavior of the probes in membrane systems will mimic the behavior of the bile acid salts. PMID- 11036530 TI - [Synthesis of cationic glucosyldiglycerides]. AB - A series of glucosyl dialkylglycerols with the pyridinium, N-morpholinium, and N methylimidazolium polar heads were synthesized by quaternization of the corresponding bases with the acetylated glucosyl diglyceride 6-O-methane- and 6-O toluenesulfonates. The resulting compounds were designed for the use in gene delivery systems. PMID- 11036531 TI - [Synthesis of amphiphilic photochromic benzo-15(18)-crown-5(6)-ethers and their properties in monolayers]. AB - New amphiphilic photochromic benzo-15(18)-crown-5(6) ethers (APC) differing in the position of the octadecyl substituent and the size of the crown cavity were synthesized. The compounds form stable monolayers in the air/water and air/alkaline metal salt solution interfaces. The results of the pressure isotherm measurements, atomic force microscopy (AFM), and electronic spectroscopy show that the structure of the monolayers formed depends on the structure of the parent APC and the nature of the cation in salt solutions. The area per molecule of APC in the monolayer (specific area) is the smallest on the water surface and increases by 20-40% on the aqueous subphase surface with an increasing concentration of salts therein to indicate the formation of APC complexes with the metal cations. When the hydrophobic aliphatic substituent is displaced from position 3 to position 5 of the benzothiazole ring, the specific area on the surface of water and subphases decreases twofold, which indicates the compactization of the monolayer on this modification. A reversible E-Z photoisomerization of APC was found in the monolayers formed in the salt solution/air interface. The features of the reaction are defined by the specific organization of the amphiphilic molecules in the monolayer and by the nature of the cation. PMID- 11036532 TI - [Phosphinic analog of methionine inhibits growth of leucosis cell L1210 and transforms to phosphinic analog of S-adenosylmethionine]. AB - A phosphinic analogue of methionine bearing a phosphinic H(OH)(O)P fragment in place of the carboxyl group inhibited the growth of the L1210 cells and was intracellularly transformed to the phosphinic analogue of S-adenosylmethionine. PMID- 11036533 TI - Mucosal biopsy techniques and interaction with the pathologist. AB - The endoscopy era made it possible to see many of the diseases that were being treated by clinicians. The use of endoscopic biopsy further enhanced that ability. This article illustrates how gastrointestinal biopsy and other practices can be improved so that patients benefit more than they might otherwise. This article focuses on pinch biopsy forceps technique and on dialogue with the pathologist. PMID- 11036534 TI - Molecular and immunologic pathology for the endoscopist. Special techniques. AB - Special techniques for the endoscopic biopsy include both immunologic (IHC) and molecular approaches (ISH, PCR). Both approaches have been adapted to foster additional diagnostic power with little or no additional work by the endoscopist. The techniques take advantage of tissue-specific antigens, unique markers in infectious diseases, and unique nucleic acid sequences present in some malignancies. From these advantages, benign conditions may be more easily distinguished from malignant ones, the causes of some infectious diseases can be confirmed, and clinically relevant classification of malignancies can be made. Some special techniques have added difficulties, such as fastidious requirements for some IHC or Southern blotting. At UCLA, approaches with the least technical challenges are those in practice for endoscopic biopsy. Lastly, the information gleaned from special techniques requires endoscopic and histopathologic context for accurate interpretation. PMID- 11036535 TI - Endoscopic mucosal resection. Current concepts. AB - Recent advances in endoscopic mucosal resection of superficial early digestive tract cancers are truly remarkable. The extraordinary long-term outcomes of patients who have been treated with endoscopic mucosal resection have encouraged the widespread practice of endoscopic mucosal resection in Japan. These minimally invasive techniques allow safe and effective treatment of diseases that would otherwise require major surgery. This article provides an overview of endoscopic mucosal resection techniques, their associated outcomes, and other potential applications of endoscopic mucosal resection. PMID- 11036536 TI - Endoscopic and endosonography guided fine-needle aspiration. AB - Endoscopic ultrasound-guided fine-needle aspiration is emerging as the preferred technique for the cytologic diagnosis of various gastrointestinal lesions. This technique may not be routinely available, but there is still a role for endoscopic or endoscopic ultrasound-assisted fine-needle aspiration. This article provides an overview of the evolution of these various techniques and discusses the advantages, disadvantages, indications, and contraindications of each. PMID- 11036538 TI - Pediatric gastrointestinal mucosal biopsy. Special considerations in children. AB - In most disorders of the gastrointestinal mucosa that occur in children and adults, the mucosal manifestations are the same. This article focuses on disorders and approaches to gastrointestinal procedures and mucosal biopsy that are of a particularly or peculiarly pediatric nature. This article discusses, in detail, the issues pertaining to endoscopy and other techniques of mucosal biopsy in children. Some approaches and techniques are considerably different than those in adults. PMID- 11036537 TI - Gastrointestinal mucosal biopsy in HIV disease and AIDS. AB - The role of the gastroenterologist as consultant for patients with HIV infection is reviewed, with a particular focus on when endoscopy with biopsy may be helpful in the diagnostic evaluation. Suggestions on where to biopsy, how to collect samples, and what pathologies might be anticipated are included. In the clinical setting of new antiviral therapies, there has been a dramatic change in the etiologic factors for common presentations such as diarrhea. A review of suspect infections and malignancies is included. PMID- 11036539 TI - Esophageal biopsy. AB - Esophageal biopsy is a natural extension of the endoscopic evaluation of many patients. It has a high likelihood of diagnostic accuracy in malignant and infectious esophageal disease. With careful attention to landmarks and good communication between the gastroenterologist and the pathologist BE should be readily diagnosed. With proper attention to the recommended biopsy protocols dysplasia should be detected accurately. Future research will determine the value of biopsy in the patient with gastroesophageal reflux and a normal endoscopy. PMID- 11036540 TI - Gastric biopsy. AB - This article focuses on the global settings where biopsy is done, the practical issues of how to do it, and what might be the benefit. The section on biopsy tips is condensed to provide very practical guidelines and some new information for even the most seasoned of endoscopists. This article covers topics such as the three histologic zones of the stomach, when to biopsy to rule out neoplasia, and biopsy in benign gastric mucosal disease. PMID- 11036541 TI - Small intestinal mucosal biopsy for investigation of diarrhea and malabsorption in adults. AB - The use of small intestinal biopsy for diagnosis in diarrhea and suspected malabsorption depends on an optimal interaction between the clinician-endoscopist and the pathologist. This necessitates open and interactive communication between involved physicians and an appreciation for correct tissue handling and biopsy orientation in the endoscopy unit and the pathology laboratory. Classification of biopsy changes on the basis of architectural abnormalities in the small intestinal biopsy may be helpful in defining the diagnosis and include severe (flat) or variably severe (mild or moderate) abnormalities. For some small intestinal disorders that are characterized by diarrhea or malabsorption, the biopsy findings may be distinctive and lead to a specific diagnosis. For others, like celiac disease, the changes are less specific, and it has become better recognized that an increasing number of conditions can produce similar histopathologic changes. Definition of typical gluten-sensitive biopsy changes in this disorder is critical. PMID- 11036542 TI - Colonoscopy plus biopsy in the inflammatory bowel diseases. AB - Colon biopsies are critical in helping to diagnose diarrhea, to distinguish different forms of colitis, to determine the extent of disease, and to determine if neoplasia has arisen in the setting of chronic colitis. Table 3 summarizes the recommended locations and numbers of biopsies for different scenarios. Some of the technical aspects pertaining to those are also discussed elsewhere in this issue. Colon biopsies in some instances can be definitive, but this usually requires the appropriate clinical scenario. For instance, to appreciate that segmental granulomatous colitis is Crohn's disease and not the much rarer colonic sarcoidosis requires ancillary clinical information. Often colon biopsies may definitively reveal an abnormality, but the findings may be nonspecific in regards to a definitive diagnosis. To use colon biopsies most appropriately in patient management and to get the most mileage from them usually requires frequent clinician-pathologist interaction, often repeat endoscopy with [table: see text] biopsies at a different time, and the assessment of the biopsies in the clinical context. PMID- 11036543 TI - [The metacarpal ligament of the thumb. Topography and functional significance of a heretofore unknown fiber tract of the thumb and its anatomic relationship to the internal interosseus muscle]. AB - During cadaver dissections of the thenar muscles of 81 human hands, we found a distinct ligament at the palmar side of the first metacarpal in 58% ([symbol: see text] 47 hands). This not previously described ligament crosses the princeps pollicis artery regularly. The ligament arises from the base of the first metacarpal bone in 83%. Sometimes (in 15%) it arises from the trapezium or from the middle portion of the first metacarpal bone in one case. The primary insertion of this ligament is the ulnar sesamoid. We have named these fibres "metacarpal ligament of the thumb". It could be classified into five different types. We think that this ligament is helpful in the stabilisation of the ulnar sesamoid. Besides it fixes the princeps pollicis artery to the metacarpal bone and therefore it is possible that it could compress the artery. Maybe the "metacarpal ligament of the thumb" is a rudiment of the interosseous muscles of the thumb or of the deep head of the flexor pollicis brevis muscle or of the oblique head of the adductor pollicis muscle. There is a very close topographic relationship between this ligament and the "internal interosseous muscle" of the thumb which was described by Schmidt and Lanz (1992) and which was mentioned by Henle (1858), who named it "M. interosseus volaris primus" for the first time. We could isolate this muscle in 69%. PMID- 11036544 TI - [Spontaneous corrections, growth disorders and post-traumatic deformities after fractures in the area of the forearm of the growing skeleton]. AB - Growth phenomena after paediatric forearm fractures are described. The capacity for spontaneous remodelling of malunions should be primarily considered in the treatment of fractures of the growing skeleton. Thus, unnecessary reductions, anaesthesia and posttraumatic deformities can be prevented. Generally speaking, a high remodelling capacity can be expected in cases of enough remaining growth, proximity to a physis with high activity, and if the main deformity lies in the plane of motion of the nearest joint. It is widely accepted that distal radius and/or ulna fractures are fully remodelled up to the age of 11 to 12 years. However, the remodelling capacity of fractures of the proximal and middle third of the shaft is smaller and less well known. Stimulating growth disturbances at the upper extremities are clinically of minor importance. Growth arrests are rare. Their fateful occurrence is not predictable and not closely related to fracture pattern or amount of dislocation. PMID- 11036545 TI - [Late sequelae of fractures of the distal third of the forearm during the growth period]. AB - Fractures to the distal third of the forearm are the most common fractures of the upper extremity, with the majority occurring between the age of ten and 14 years. With the exception of the rare epiphyseal fractures, they have a favourable prognosis. The present study investigates the frequency and extent of potential clinical and radiological late sequelae of fractures in the distal third of the forearm during growth. Of the patients treated at the Innsbruck University Department of Traumatology from 1980 to 1992, 220 patients of a growing age with 232 closed fractures in the distal third of the forearm were followed up. The radius alone was affected in 60% of these cases; the radius and the ulna in 40%. Fractures of the ulna alone were not present. The mean age of the patients at the time of injury was nine years (range one to 16 years) and the mean time of follow up ten years (range five to 16 years). In addition to the patient's subjective assessment, the right and left sides were compared with regard to mobility of the wrist and rotational movement of the forearm. Based on standard X-rays, the frontal (radio-ulnar) and lateral (dorso-palmar) radial joint angle as well as the difference in the radio-ulnar plane were compared with the contralateral side. Clinical and radiological findings were summarised into an overall result. 19% of the patients reported pain in the injured wrist. Mobility of the wrist in the sagittal and/or frontal plane was limited in 5% of patients and rotation of the forearm was limited in 16% of patients. A statistically significant accumulation of limited rotation was seen after physeal fractures of the ulna ("one-way" ANOVA-test, p = 0.0033). A difference between the left and right side in regard to the frontal radial joint angle was seen in 6% of patients and a difference in the lateral radial joint angle was registered in 2% of patients. A difference in the radio-ulnar plane was observed in 37% of patients. In the presence of relative ulna-plus variance, 75% of patients complained of pain in the ulnocarpal compartment of the wrist. In these patients, dynamic magnetic resonance tomography revealed a compression of the ulnocarpal disk between the proximal carpal bones and the head of the ulna, as well as degeneration in the central portion of the disk. The overall outcome was very good in 72%, good in 19%, moderate in 6% and poor in 3% of patients. The younger the children had been at the time of injury, the more favourable were the results (chi-square test, p = 0.009). Children older than ten years of age with an angulatory deformity of more than 20 degrees and/or fragment dislocation over half of the breadth of the shaft at fracture consolidation showed the poorest results. Further factors having a negative influence on the outcome were repeated reduction manoeuvres and an additional fracture of the ulna. PMID- 11036546 TI - [Malunited fractures of the forearm during the growth period with special reference to the forearm longitudinal axis. Case reports]. AB - Malunion of forearm fractures located in the distal third will remodel satisfactorily providing the child is less than twelve years of age. Complete correction of gross deformity cannot be anticipated in diaphysial fractures when the child is over five years of age. Malunion of fractures of the forearm in children can lead to permanent functional disability with limitation of forearm rotation. In the case of functional disability, there is an indication for corrective osteotomy at the age over twelve in malunion of a fracture located in the distal third and already after age five in gross deformity of fractures to the midshaft of the forearm. PMID- 11036547 TI - [Monteggia injuries in childhood]. AB - The Monteggia-fracture dislocation is a characteristic combined injury of the forearm with fracture of the ulna and dislocation of the head of the radius. Monteggia- and equivalent injuries are rare. In the present study, we attempt to clarify the concept of therapy and observe the influence of this injury on the growth of the forearm bones. Between 1977 and 1996, 27 patients (three to thirteen years) with a Monteggia or a Monteggia-equivalent injury were treated. We present clinical and radiological long-term results of 20 patients, two to 21 years following the injury. Group I includes 12 patients with a classic Monteggia injury, five patients were treated conservatively, seven patients surgically. Ten patients were free of pain and had no loss of motion. In one patient there was a loss of pronation following an accompanying injury of the distal radius. Primary paresis of the radial nerve has a good prognosis. One patient presents a persisting dislocation of the radius head with a loss of flexion in the elbow joint, and a secondary persisting paraesthesia of the median nerve. Group II includes eight patients with a Monteggia-equivalent injury; all of them were treated surgically. The results in this group showed more loss of motion in elbow function, forearm rotation, and dislocation of the axis in the elbow joint. Monteggia and Monteggia-equivalent injuries in childhood have good functional results if correct reduction of the fracture of the ulna and the head of the radius is performed. If this is not possible conservatively, patients need open reduction and internal fixation. PMID- 11036548 TI - [Percutaneous osteosynthesis of scaphoid fracture with the Herbert-Whipple screw- technique and results]. AB - In order to diminish the surgical trauma a percutaneous osteosynthesis of acute fractures of the scaphoid was performed in 22 cases using the cannulated Herbert Whipple screw. Displaced fractures were reduced under fluoroscopy in a position of extension and supination while pulling the thumb with a weight of 5 kg by means of a Japanese finger trap. Additional correction could be achieved by using Kirschner wires as joysticks. The guide wire allowed a correct positioning of the cannulated screw. We have found that radiocarpal and midcarpal arthroscopy was of little benefit for fracture reduction. However, arthroscopy was important for the detection of concomitant carpal injuries. Due to minor projection over the distal scaphoid pole, three screws had to be removed underlining the difficulty of exact length determination. Sound bony union was achieved in all 22 cases even in those treated without plaster immobilisation. Limitations in wrist motion and grip strength have been found after concomitant injuries such as fractures of the distal radius and scapholunate ligament injury. On the other hand, there was no impairment of wrist function after percutaneous osteosynthesis of isolated scaphoid fractures. In conclusion, percutaneous osteosynthesis of the fractured scaphoid with the cannulated Herbert-Whipple screw permits a reliable treatment with minimal operative trauma. PMID- 11036549 TI - [Use of a cannulated 3.0 mm AO screw with an intraosseous support washer in osteosynthesis of the scaphoid: results and analysis of problems in 28 cases]. AB - The cannulated 3.0 mm AO/ASIF screw with threaded washer is a new implant for scaphoid osteosynthesis. After insertion into the distal scaphoid pole, the washer serves as an intraosseous support for the head of the cannulated lag screw. From June 1997 to March 1998 the new implant was used in 28 male patients between 14 and 50 years of age. In fourteen patients, acute scaphoid fractures were operated on, including five transscaphoid perilunate fracture dislocations (Herbert type B2 and B4, respectively). Fourteen patients had scaphoid pseudarthroses, among those two re-pseudarthroses after prior surgery elsewhere. Using a palmar approach, the scaphoid was reduced. In pseudarthroses an iliac cortico-cancellous bone block was inserted, and the implants were inserted in a distal-to-proximal manner. In the fourteen acute fractures, follow-up assessment at a mean of 11 months revealed one pseudarthrosis (7%) and four screw removals (29%) for screw back-out after bony consolidation. In the fourteen pseudarthroses, follow-up assessment at a mean of ten months revealed one re pseudarthrosis (7%) after a technical fault. It was treated by re-operation using the same implant, and healing was now uneventful. One screw removal became necessary after bony consolidation in another patient (7%). In conclusion, our preliminary results suggest that the new implant is suitable for stabilization of scaphoid pseudarthroses after insertion of an iliac crest bone graft. The complication rate in the treatment of acute fractures was inacceptably high. PMID- 11036550 TI - [Malignant course of pigmented villonodular synovitis of the flexor tendon sheath of the small finger--case report and review of the literature]. AB - The case of a 55-year-old patient is described, presenting the clinical sign of a chronic infection of the tendon sheath. Upon incision a mass of brown synovial tumor was found. The X-ray showed a circumscript bone erosion. MRI demonstrated tumor involvement of all flexor tendons up to the forearm. Under radiation the tumor initially was diminished. Half a year later, pulmonary metastases were found. The destruction of the whole skeleton of the hand led to a forearm amputation. Later, a metastasis was found in the tongue. The patient died with the clinical signs of pulmonary insufficiency. Autopsy showed diffuse pulmonary metastases. This case is discussed together with other rare cases of malignant pigmented villonodular synovialitis arising from joints and tendon sheaths. PMID- 11036551 TI - Computational modeling of left heart diastolic function: examination of ventricular dysfunction. AB - A computational model that accounts for blood-tissue interaction under physiological flow conditions was developed and applied to a thin-walled model of the left heart. This model consisted of the left ventricle, left atrium, and pulmonary vein flow. The input functions for the model included the pulmonary vein driving pressure and time-dependent relationship for changes in chamber tissue properties during the simulation. The Immersed Boundary Method was used for the interaction of the tissue and blood in response to fluid forces and changes in tissue pathophysiology, and the fluid mass and momentum conservation equations were solved using Patankar's Semi-Implicit Method for Pressure Linked Equations (SIMPLE). This model was used to examine the flow fields in the left heart under abnormal diastolic conditions of delayed ventricular relaxation, delayed ventricular relaxation with increased ventricular stiffness, and delayed ventricular relaxation with an increased atrial contraction. The results obtained from the left heart model were compared to clinically observed diastolic flow conditions, and to the results from simulations of normal diastolic function in this model [1]. Cases involving impairment of diastolic function were modeled with changes to the input functions for fiber relaxation/contraction of the chambers. The three cases of diastolic dysfunction investigated agreed with the changes in diastolic flow fields seen clinically. The effect of delayed relaxation was to decrease the early filling magnitude, and this decrease was larger when the stiffness of the ventricle was increased. Also, increasing the contraction of the atrium during atrial systole resulted in a higher late filling velocity and atrial pressure. The results show that dysfunction can be modeled by changing the relationships for fiber resting-length and/or stiffness. This provides confidence in future modeling of disease, especially changes to chamber properties to examine the effect of local dysfunction on global flow fields. PMID- 11036552 TI - Observation and quantification of gas bubble formation on a mechanical heart valve. AB - Clinical studies using transcranial Doppler ultrasonography in patients with mechanical heart valves (MHV) have detected gaseous emboli. The relationship of gaseous emboli release and cavitation on MHV has been a subject of debate in the literature. To study the influence of cavitation and gas content on the formation and growth of stable gas bubbles, a mock circulatory loop, which employed a Medtronic-Hall pyrolytic carbon disk valve in the mitral position, was used. A high-speed video camera allowed observation of cavitation and gas bubble release on the inflow valve surfaces as a function of cavitation intensity and carbon dioxide (CO2) concentration, while an ultrasonic monitoring system scanned the aortic outflow tract to quantify gas bubble production by calculating the gray scale levels of the images. In the absence of cavitation, no stable gas bubbles were formed. When gas bubbles were formed, they were first seen a few milliseconds after and in the vicinity of cavitation collapse. The volume of the gas bubbles detected in the aortic track increased with both increased CO2 and increased cavitation intensity. No correlation was observed between O2 concentration and bubble volume. We conclude that cavitation is an essential precursor to stable gas bubble formation, and CO2, the most soluble blood gas, is the major component of stable gas bubbles. PMID- 11036553 TI - Physiological flow simulation in residual human stenoses after coronary angioplasty. AB - To evaluate the local hemodynamic implications of coronary artery balloon angioplasty, computational fluid dynamics (CFD) was applied in a group of patients previously reported by [Wilson et al. (1988), 77, pp. 873-885] with representative stenosis geometry post-angioplasty and with measured values of coronary flow reserve returning to a normal range (3.6 +/- 0.3). During undisturbed flow in the absence of diagnostic catheter sensors within the lesions, the computed mean pressure drop delta p was only about 1 mmHg at basal flow, and increased moderately to about 8 mmHg for hyperemic flow. Corresponding elevated levels of mean wall shear stress in the midthroat region of the residual stenoses, which are common after angioplasty procedures, increased from about 60 to 290 dynes/cm2 during hyperemia. The computations (Ree approximately equal to 100-400; alpha e = 2.25) indicated that the pulsatile flow field was principally quasi-steady during the cardiac cycle, but there was phase lag in the pressure drop-mean velocity (delta p - u) relation. Time-averaged pressure drop values, delta p, were about 20 percent higher than calculated pressure drop values, delta ps, for steady flow, similar to previous in vitro measurements by Cho et al. (1983). In the throat region, viscous effects were confined to the near-wall region, and entrance effects were evident during the cardiac cycle. Proximal to the lesion, velocity profiles deviated from parabolic shape at lower velocities during the cardiac cycle. The flow field was very complex in the oscillatory separated flow reattachment region in the distal vessel where pressure recovery occurred. These results may also serve as a useful reference against catheter measured pressure drops and velocity ratios (hemodynamic endpoints) and arteriographic (anatomic) endpoints post-angioplasty. Some comparisons to previous studies of flow through stenoses models are also shown for perspective purposes. PMID- 11036554 TI - Hemolytic potential of hydrodynamic cavitation. AB - The purpose of this study was to determine the hemolytic potentials of discrete bubble cavitation and attached cavitation. To generate controlled cavitation events, a venturigeometry hydrodynamic device, called a Cavitation Susceptibility Meter (CSM), was constructed. A comparison between the hemolytic potential of discrete bubble cavitation and attached cavitation was investigated with a single pass flow apparatus and a recirculating flow apparatus, both utilizing the CSM. An analytical model, based on spherical bubble dynamics, was developed for predicting the hemolysis caused by discrete bubble cavitation. Experimentally, discrete bubble cavitation did not correlate with a measurable increase in plasma free hemoglobin (PFHb), as predicted by the analytical model. However, attached cavitation did result in significant PFHb generation. The rate of PFHb generation scaled inversely with the Cavitation number at a constant flow rate, suggesting that the size of the attached cavity was the dominant hemolytic factor. PMID- 11036555 TI - Biaxial mechanical properties of the native and glutaraldehyde-treated aortic valve cusp: Part II--A structural constitutive model. AB - We have formulated the first constitutive model to describe the complete measured planar biaxial stress-strain relationship of the native and glutaraldehyde treated aortic valve cusp using a structurally guided approach. When applied to native, zero-pressure fixed, and low-pressure fixed cusps, only three parameters were needed to simulate fully the highly anisotropic, and nonlinear in-plane biaxial mechanical behavior. Differences in the behavior of the native and zero- and low-pressure fixed cusps were found to be primarily due to changes in the effective fiber stress-strain behavior. Further, the model was able to account for the effects of small (< 10 deg) misalignments in the cuspal specimens with respect to the biaxial test axes that increased the accuracy of the model material parameters. Although based upon a simplified cuspal structure, the model underscored the role of the angular orientation of the fibers that completely accounted for extreme mechanical anisotropy and pronounced axial coupling. Knowledge of the mechanics of the aortic cusp derived from this model may aid in the understanding of fatigue damage in bioprosthetic heart valves and, potentially, lay the groundwork for the design of tissue-engineered scaffolds for replacement heart valves. PMID- 11036556 TI - On the electric potentials inside a charged soft hydrated biological tissue: streaming potential versus diffusion potential. AB - The main objective of this study is to determine the nature of electric fields inside articular cartilage while accounting for the effects of both streaming potential and diffusion potential. Specifically, we solve two tissue mechano electrochemical problems using the triphasic theories developed by Lai et al. (1991, ASME J. Biomech Eng., 113, pp. 245-258) and Gu et al. (1998, ASME J. Biomech. Eng., 120, pp. 169-180) (1) the steady one-dimensional permeation problem; and (2) the transient one-dimensional ramped-displacement, confined compression, stress-relaxation problem (both in an open circuit condition) so as to be able to calculate the compressive strain, the electric potential, and the fixed charged density (FCD) inside cartilage. Our calculations show that in these two technically important problems, the diffusion potential effects compete against the flow-induced kinetic effects (streaming potential) for dominance of the electric potential inside the tissue. For softer tissues of similar FCD (i.e., lower aggregate modulus), the diffusion potential effects are enhanced when the tissue is being compressed (i.e., increasing its FCD in a nonuniform manner) either by direct compression or by drag-induced compaction; indeed, the diffusion potential effect may dominate over the streaming potential effect. The polarity of the electric potential field is in the same direction of interstitial fluid flow when streaming potential dominates, and in the opposite direction of fluid flow when diffusion potential dominates. For physiologically realistic articular cartilage material parameters, the polarity of electric potential across the tissue on the outside (surface to surface) may be opposite to the polarity across the tissue on the inside (surface to surface). Since the electromechanical signals that chondrocytes perceive in situ are the stresses, strains, pressures and the electric field generated inside the extracellular matrix when the tissue is deformed, the results from this study offer new challenges for the understanding of possible mechanisms that control chondrocyte biosyntheses. PMID- 11036557 TI - A microstructural model of elastostatic properties of articular cartilage in confined compression. AB - A microstructural model of cartilage was developed to investigate the relative contribution of tissue matrix components to its elastostatic properties. Cartilage was depicted as a tensed collagen lattice pressurized by the Donnan osmotic swelling pressure of proteoglycans. As a first step in modeling the collagen lattice, two-dimensional networks of tensed, elastic, interconnected cables were studied as conceptual models. The models were subjected to the boundary conditions of confined compression and stress-strain curves and elastic moduli were obtained as a function of a two-dimensional equivalent of swelling pressure. Model predictions were compared to equilibrium confined compression moduli of calf cartilage obtained at different bath concentrations ranging from 0.01 to 0.50 M NaCl. It was found that a triangular cable network provided the most consistent correspondence to the experimental data. The model showed that the cartilage collagen network remained tensed under large confined compression strains and could therefore support shear stress. The model also predicted that the elastic moduli increased with increasing swelling pressure in a manner qualitatively similar to experimental observations. Although the model did not preclude potential contributions of other tissue components and mechanisms, the consistency of model predictions with experimental observations suggests that the cartilage collagen network, prestressed by proteoglycan swelling pressure, plays an important role in supporting compression. PMID- 11036558 TI - In vivo modeling of interstitial pressure in the brain under surgical load using finite elements. AB - Current brain deformation models have predominantly reflected solid constitutive relationships generated from empirical ex vivo data and have largely overlooked interstitial hydrodynamic effects. In the context of a technique to update images intraoperatively for image-guided neuronavigation, we have developed and quantified the deformation characteristics of a three-dimensional porous media finite element model of brain deformation in vivo. Results have demonstrated at least 75-85 percent predictive capability, but have also indicated that interstitial hydrodynamics are important. In this paper we investigate interstitial pressure transient behavior in brain tissue when subjected to an acute surgical load consistent with neurosurgical events. Data are presented from three in vivo porcine experiments where subsurface tissue deformation and interhemispheric pressure gradients were measured under conditions of an applied mechanical deformation and then compared to calculations with our three dimensional brain model. Results demonstrate that porous-media consolidation captures the hydraulic behavior of brain tissue subjected to comparable surgical loads and that the experimental protocol causes minimal trauma to porcine brain tissue. Working values for hydraulic conductivity of white and gray matter are also reported and an assessment of transient pressure gradient effects with respect to deformation is provided. PMID- 11036559 TI - Infant skull and suture properties: measurements and implications for mechanisms of pediatric brain injury. AB - The mechanical properties of the adult human skull are well documented, but little information is available for the infant skull. To determine the age dependent changes in skull properties, we tested human and porcine infant cranial bone in three-point bending. The measurement of elastic modulus in the human and porcine infant cranial bone agrees with and extends previous published data [McPherson, G. K., and Kriewall, T. J. (1980), J. Biomech., 13, pp. 9-16] for human infant cranial bone. After confirming that the porcine and human cranial bone properties were comparable, additional tensile and three-point bending studies were conducted on porcine cranial bone and suture. Comparisons of the porcine infant data with previously published adult human data demonstrate that the elastic modulus, ultimate stress, and energy absorbed to failure increase, and the ultimate strain decreases with age for cranial bone. Likewise, we conclude that the elastic modulus, ultimate stress, and energy absorbed to failure increase with age for sutures. We constructed two finite element models of an idealized one-month old infant head, one with pediatric and the other adult skull properties, and subjected them to impact loading to investigate the contribution of the cranial bone properties on the intracranial tissue deformation pattern. The computational simulations demonstrate that the comparatively compliant skull and membranous suture properties of the infant brain case are associated with large cranial shape changes, and a more diffuse pattern of brain distortion than when the skull takes on adult properties. These studies are a fundamental initial step in predicting the unique mechanical response of the pediatric skull to traumatic loads associated with head injury and, thus, for defining head injury thresholds for children. PMID- 11036560 TI - Studies on the three-dimensional temperature transients in the canine prostate during transurethral microwave thermal therapy. AB - Thermal therapy of benign prostatic hyperplasia requires accurate prediction of the temperature distribution induced by the heating within the prostatic tissue. In this study, the Pennes bioheat transfer equation was used to model the transient heat transfer inside the canine prostate during transurethral microwave thermal therapy. Incorporating the specific absorption rate of microwave energy in tissue, a closed-form analytical solution was obtained. Good agreement was found between the theoretical predictions and in-vivo experimental results. Effects of blood perfusion and the cooling at the urethral wall on the temperature rise were investigated within the prostate during heating. The peak intraprostatic temperatures attained by application of 5, 10, or 15 W microwave power were predicted to be 38 degrees C, 41 degrees C, and 44 degrees C. Results from this study will help optimize the thermal dose that can be applied to target tissue during the therapy. PMID- 11036561 TI - Predictions of mechanical output of the human M. triceps surae on the basis of electromyographic signals: the role of stimulation dynamics. AB - In order to assess the significance of the dynamics of neural control signals for the rise time of muscle moment, simulations of isometric and dynamic plantar flexion contractions were performed using electromyographic signals (EMG signals) of m. triceps surae as input. When excitation dynamics of the muscle model was optimized for an M-wave of the medial head of m. gastrocnemius (GM), the model was able to make reasonable predictions of the rise time of muscle moment during voluntary isometric plantar flexion contractions on the basis of voluntary GM EMG signals. The rise time of muscle moment in the model was for the greater part determined by the amplitude of the first EMG burst. For dynamic jumplike movements of the ankle joint, however, no relationship between rise time of muscle moment in the experiment and muscle moment predicted by the model on the basis of GM EMG signals was found. Since rise time of muscle moment varied over a small range for this movement, it cannot be completely excluded that stimulation dynamics plays a role in control of these simple single-joint movements. PMID- 11036562 TI - Substrate deformation levels associated with routine physical activity are less stimulatory to bone cells relative to loading-induced oscillatory fluid flow. AB - Although it is well accepted that bone tissue metabolism is regulated by external mechanical loads, it remains unclear to what load-induced physical signals bone cells respond. In this study, a novel computer-controlled stretch device and parallel plate flow chamber were employed to investigate cytosolic calcium (Ca2+i) mobilization in response to a range of dynamic substrate strain levels (0.1-10 percent, 1 Hz) and oscillating fluid flow (2 N/m2, 1 Hz). In addition, we quantified the effect of dynamic substrate strain and oscillating fluid flow on the expression of mRNA for the bone matrix protein osteopontin (OPN). Our data demonstrate that continuum strain levels observed for routine physical activities (< 0.5 percent) do not induce Ca2+i responses in osteoblastic cells in vitro. However, there was a significant increase in the number of responding cells at larger strain levels. Moreover, we found no change in osteopontin mRNA level in response to 0.5 percent strain at 1 Hz. In contrast, oscillating fluid flow predicted to occur in the lacunar-canalicular system due to routine physical activities (2 N/m2, 1 Hz) caused significant increases in both Ca2+i and OPN mRNA. These data suggest that, relative to fluid flow, substrate deformation may play less of a role in bone cell mechanotransduction associated with bone adaptation to routine loads. PMID- 11036563 TI - Cell-level finite element studies of viscous cells in planar aggregates. AB - A new cell-level finite element formulation is presented and used to investigate how epithelia and other planar collections of viscous cells might deform during events such as embryo morphogenesis and wound healing. Forces arising from cytoskeletal components, cytoplasm viscosity, and cell-cell adhesions are included. Individual cells are modeled using multiple finite elements, and cell rearrangements can occur. Simulations of cell-sheet stretching indicate that the initial stages of sheet stretching are characterized by changes in cell shape, while subsequent stages are governed by cell rearrangement. Inferences can be made from the simulations about the forces that act in real cell sheets when suitable experimental data are available. PMID- 11036564 TI - The mechanics of heterotypic cell aggregates: insights from computer simulations. AB - Finite element-based computer simulations are used to investigate a number of phenomena, including tissue engulfment, cell sorting, and checkerboard-pattern formation, exhibited by heterotypic cell aggregates. The simulations show that these phenomena can be driven by a single equivalent force, namely a surface (or interfacial) tension, that results from cytoskeletal components and cell-cell adhesions. They also reveal that tissue engulfment, cell sorting, and checkerboard-pattern formation involve several discernible mechanical features or stages. With the aid of analytical arguments, we identify the conditions necessary for each of these phenomena. These findings are consistent with previous experimental investigations and computer simulations, but pose significant challenges to current theories of cell sorting and tissue engulfment. PMID- 11036565 TI - Asymptotic analysis of the stress field in adhering dental restorations. AB - Polymer-based composites are widely used in restorative dentistry as alternatives to metals and ceramics to fill cavities in teeth. They adhere to the walls of the cavity in the tooth, thus forming a composite body consisting of dentine, enamel, and composite resin. Geometric discontinuities along the interfaces between these materials can induce singularities in the stress field, which in turn lead to premature failure of the restoration. In the present investigation, a complex stress function technique is employed to derive the order of the stress singularity. It is shown that the order of the singularity depends on both the material properties of the restorative material and the local geometry of the cavity. It is also shown that the singularity in the stress field can be avoided through careful design of the cavity shape. The results presented correlate well with experimental results reported in the literature. PMID- 11036566 TI - Bending vibrations of the femur and the oscillatory behavior of a cemented femoral hip endoprosthesis. AB - The paper presents a novel method for recording amplitude and phase of 6D vibrations of a spatial pendulum over a wide frequency range (10 Hz up to 20 kHz). The six degrees of freedom of the pendulum mass were monitored by three electrodynamic stereo pickups. At rest, the tips of the needles and the pendulum's center of mass defined the reference system with respect to which the oscillations of the mass were recorded in terms of their amplitudes and phases. Its small dimensions, constant transfer characteristics, linearity, high dynamics, and virtual lack of reaction onto the moving system over the entire frequency range provided the advantages of the measuring system. This method was used to analyze the spatial 6D-vibrations of the head of a cemented femoral hip endoprosthesis when the femur was stimulated to bending vibrations. The head of the prosthesis carried out axial rotational vibrations at every frequency used to stimulate the femur. The amplitudes of the axial rotations of the cortical bone were small in comparison to the ones of the prosthesis head, indicating that axial rotational vibrations following femur bending vibrations mainly stressed the spongiosa and the cement layer. This was observed over the entire frequency range, including at the low frequencies relevant for gait. Over the low-frequency range, as well as at some of the higher resonance frequencies, stationary instantaneous helical axes characterized the vibrations. The measurements suggest the mechanism that the interface "implant-bone" may already be stressed by axial torsional loads when the femur is loaded by bending impacts that are known to occur during walking. PMID- 11036567 TI - Validity of photoelastic strain measurement on cadaveric proximal femora. AB - Rosette strain gages indicate shear and principal strains at specific points, whereas photoelastic coatings provide shear strain information over a broad area. Information regarding bone loading and load transfer from a prosthetic implant to adjacent bone can be obtained using either strain-measuring technique on loaded femora. This study compared proximal femoral strains derived from photoelastic coatings to those obtained from rosette strain gages applied directly to the bone in order to determine the relationships between photoelastic shear strains and rosette shear and principal strains. Photoelastic shear strains underestimated rosette shear strains and exceeded the larger of the rosette principal strains. Principal strains derived from photoelastic coatings augmented with strain separator gages underestimated their rosette counterparts in most instances. Correlation was strong and nearly linear for all measures, indicating that photoelastic coatings can accurately express proportional strain changes despite imperfect agreement in absolute strain magnitudes. The best agreement between absolute strain magnitudes occurred in the proximal medial, or calcar, region. Understanding the relationships between the various measures obtained using the two strain measurement methods will allow more accurate estimates of actual strains to be made from photoelastic coatings. PMID- 11036568 TI - Three-dimensional finite element analysis of glenoid replacement prostheses: a comparison of keeled and pegged anchorage systems. AB - Glenoid component loosening is the dominant cause of failure in total shoulder arthroplasty. It is presumed that loosening in the glenoid is caused by high stresses in the cement layer. Several anchorage systems have been designed with the aim of reducing the loosening rate, the two major categories being "keeled" fixation and "pegged" fixation. However, no three-dimensional finite element analysis has been performed to quantify the stresses in the cement or to compare the different glenoid prosthesis anchorage systems. The objective of this study was to determine the stresses in the cement layer and surrounding bone for glenoid replacement components. A three-dimensional model of the scapula was generated using CT data for geometry and material property definition. Keeled and pegged designs were inserted into the glenoid, surrounded by a 1-mm layer of bone cement. A 90 deg arm abduction load with a full muscle and joint load was applied, following van der Helm (1994). Deformations of the prosthesis, stresses in the cement, and stresses in the bone were calculated. Stresses were also calculated for a simulated case of rheumatoid arthritis (RA) in which bone properties were modified to reflect that condition. A maximum principal stress based failure model was used to predict what quantity of the cement is at risk of failure at the levels of stress computed. The prediction is that 94 percent (pegged prosthesis) and 68 percent (keeled prosthesis) of the cement has a greater than 95 percent probability of survival in normal bone. In RA bone, however, the situation is reversed where 86 percent (pegged prosthesis) and 99 percent (keeled prosthesis) of the cement has a greater than 95 percent probability of survival. Bone stresses are shown to be not much affected by the prosthesis design, except at the tip of the central peg or keel. It is concluded that a "pegged" anchorage system is superior for normal bone, whereas a "keeled" anchorage system is superior for RA bone. PMID- 11036569 TI - A well-posed, embedded constraint representation of joint moments from kinesiological measurements. AB - Joint moment estimation using the traditional inverse dynamics analysis presents two challenging problems, which limit its reliability. First, the quality of the computed moments depends directly on unreliable estimates of the segment accelerations obtained numerically by differentiating noisy marker measurements. Second, the representation of joint moments from combined video and force plate measurements belongs to a class of ill-posed problems, which does not possess a unique solution. This paper presents a well-posed representation derived from an embedded constraint equation. The proposed method, referred to as the embedded constraint representation (ECR), provides unique moment estimates, which satisfy all measurement constraints and boundary conditions and require fewer acceleration components than the traditional inverse dynamics method. Specifically, for an n-segment open chain planar system, the ECR requires n-3 acceleration components as compared to 3(n-1) components required by the traditional (from ground up) inverse dynamics analysis. Based on a simulated experiment using a simple three-segment model, the precision of the ECR is evaluated at different noise levels and compared to the traditional inverse dynamics technique. At the lowest noise levels, the inverse dynamics method is up to 50 percent more accurate while at the highest noise levels the ECR method is up to 100 percent more accurate. The ECR results over the entire range of noise levels reveals an average improvement on the order 20 percent in estimating the moments distal to the force plate and no significant improvement in estimating moments proximal to the force plate. The new method is particularly advantageous in a combined video, force plate, and accelerometery sensing strategy. PMID- 11036570 TI - Bicycle drive system dynamics: theory and experimental validation. AB - Bicycle pedaling has been studied from both a motor control and an equipment setup and design perspective. In both cases, although the dynamics of the bicycle drive system may have an influence on the results, a thorough understanding of the dynamics has not been developed. This study pursued three objectives related to developing such an understanding. The first was to identify the limitations of the inertial/frictional drive system model commonly used in the literature. The second was to investigate the advantages of an inertial/frictional/compliant model. The final objective was to use these models to develop a methodology for configuring a laboratory ergometer to emulate the drive system dynamics of road riding. Experimental data collected from the resulting road-riding emulator and from a standard ergometer confirmed that the inertial/frictional model is adequate for most studies of road-riding mechanics or pedaling coordination. However, the compliant model was needed to reproduce the phase shift in crank angle variations observed experimentally when emulating the high inertia of road riding. This finding may be significant for equipment setup and design studies where crank kinematic variations are important or for motor control studies where fine control issues are of interest. PMID- 11036571 TI - The osmotic swelling characteristics of cardiac valve prostheses. AB - Several types of mechanical cardiac prostheses have been constructed with Delrin occluders, a material that is subject to osmotic swelling. The leaftets are designed to expand to specific tolerances when immersed in blood. The synthetic blood analogs commonly used in vitro contain hydrophilic compounds that can alter the osmotic expansion of the Delrin occluders. A static leak test chamber was employed to illustrate the effects of various test fluids on the sustained regurgitation phase of Delrin valves. PMID- 11036572 TI - Effect of residual stress and heterogeneity on circumferential stress in the arterial wall. AB - Quantifying the stress distribution through the arterial wall is essential to studies of arterial growth and disease. Previous studies have shown that both residual stress, as measured by opening angle, and differing material properties for the media-intima and the adventitial layers affect the transmural circumferential stress (sigma theta) distribution. Because a lack of comprehensive data on a single species and artery has led to combinations from multiple sources, this study determined the sensitivity of sigma theta to published variations in both opening angle and layer thickness data. We fit material properties to previously published experimental data for pressure diameter relations and opening angles of rabbit carotid artery, and predicted sigma theta through the arterial wall at physiologic conditions. Using a one layer model, the ratio of sigma theta at the internal wall to the mean sigma theta decreased from 2.34 to 0.98 as the opening angle increased from 60 to 130 deg. In a two-layer model using a 95 deg opening angle, mean sigma theta in the adventitia increased (112 percent for 25 percent adventitia) and mean sigma theta in the media decreased (47 percent for 25 percent adventitia). These results suggest that both residual stress and wall layers have important effects on transmural stress distribution. Thus, experimental measurements of loading curves, opening angles, and wall composition from the same species and artery are needed to accurately predict the transmural stress distribution in the arterial wall. PMID- 11036573 TI - A system for quantifying the cooling effectiveness of bicycle helmets. AB - This article describes the design and development of a system that is capable of quantifying the thermal comfort of bicycle helmets. The motivation for the development of the system stems from the desire both to increase helmet use and to provide the designer with a quantitative method of evaluating the thermal comfort of a helmet. The system consists of a heated mannequin head form, a heated reference sphere, a small wind tunnel, and a data acquisition system. Both the head form and the reference sphere were instrumented with thermocouples. The system is capable of simulating riding speeds ranging from 4.5-15.5 m/s. A cooling effectiveness, C1, that is independent of both ambient conditions and wind velocity is defined as a measure of how well the helmet ventilates as compared to the reference sphere. The system was validated by testing six commercially available bicycle helmets manufactured between approximately 1992 and 1998. PMID- 11036574 TI - Comparison of cardiotropic drug effects on haemodynamic and myocardial energetics in patients with heart failure: a computer simulation. AB - The purpose of our study is to compare haemodynamic responses and the ischaemic potential of commonly used inotropes (dopamine, dobutamine and milrinone) using a computer model of the cardiovascular system. Cardiotropic drugs interact with the model by changing ventricular elastance and resistance of the individual circulation. All three drugs increase cardiac index in a dose-dependent manner. Dopamine at medium and high infusion rates increases heart rate, systemic vascular resistance and arterial blood pressure. The associated increase in coronary blood flow, however, is not sufficient to account for increased oxygen demand. Both dobutamine and milrinone decrease vascular resistance and increase coronary blood flow. The more pronounced increase in heart rate associated with dobutamine, however, results in a higher ischaemic potential for this drug. Our simulation demonstrates that although all the drugs studied improve cardiac function in simulated patients with heart failure, milrinone accomplishes this at a lower energy cost. The computer simulation developed can be used to assess the complex effect of cardiotropic drugs and possibly suggest optimal drug therapy in specific clinical situations. PMID- 11036575 TI - Different responses of Finapres and the oscillometric finger blood pressure monitor during intensive vasomotion. AB - Two different techniques for non-invasive beat-to-beat finger arterial blood pressure monitoring are compared in six healthy volunteers during local hand heating from 21 to 38 degrees C. The degree of peripheral vasoconstriction was established by recording the thumb pulp skin blood flow with a laser Doppler instrument. For time episodes without vasoconstriction no systematic difference in the readings of beat-to-beat mean blood pressure of the two monitors was found (the oscillometric device UT9201 minus Finapres difference was 0.3 mm Hg, SD 0.3). For the episodes with vasoconstriction the difference was statistically significant (6.7 mm Hg, SD 2.0). The oscillometric device minus Finapres difference and the laser Doppler signal were found to be inversely correlated, the correlation coefficient varying from -0.28 to -0.67. A disagreement between the readings of the instruments during intensive vasomotion is assumed to be caused mainly by the tendency of the oscillometric method to overestimate the finger mean blood pressure under the condition of peripheral vasoconstriction. PMID- 11036576 TI - Computational haemodynamics analysis and comparison study of arterio-venous grafts. AB - Haemodialysis arterio-venous graft failure is related to the development of stenotic lesions most commonly located near the venous anastomosis, especially in the toe region. 'Disturbed flow' interaction with the vessel wall surface, characterized by haemodynamic parameters based on the local wall shear stress or the radial pressure gradient, have been widely recognized as the trigger mechanism of a cascade of abnormal biological events leading to occlusive developments. Assuming incompressible laminar flow and rigid, in-plane vessel walls, validated haemodynamics are numerically simulated for a constant-diameter end-to-side base case, the Venaflo graft, and an improved graft-end configuration. The geometric design of the new graft-end was based on the reduction of three time- and area-averaged haemodynamic parameters, i.e. the wall shear stress gradient, wall shear stress angle gradient, and radial pressure gradient. Considering the critical toe region, the Venaflo graft has demonstrated measurable improvements over the base case configuration in predictive computer simulations as well as in clinical trials. The performance improvement should be further enhanced with the modifications illustrated by the new design. PMID- 11036577 TI - A virtual instrument (VI) for haemodynamic management in ICU and during surgery. AB - Systolic pressure variations (SPV) during mechanical ventilation and its single components, related to short apnea, reflect changes of the volemic condition of the patient. To introduce their determination during clinical monitoring for different fluid states and for different tidal volumes, they must be computed on line without introducing interference with standard activities. A system computing on-line systolic pressure variation during mechanical ventilation, connected to standard monitoring devices, has been proposed. It is based on a notebook PC implemented with graphical software comprising a user panel in the form of a virtual instrument and is able to acquire, process and present signals from different instruments utilized in ICU and during surgery. It can be used as a base to assess the ability of computed parameters helpful in clinical decision. The use of a notebook PC and open software allows operators, even if non-expert in computer science, to test and implement this, as well as other innovative tools in clinical practice. PMID- 11036578 TI - Q-Pro: a quality control management system for medical equipment. AB - Q-Pro is an application for quality control (QC) and inspection of medical equipment. The system has been designed on the basis of a broad requirements analysis, contributed by clinical engineers from several European countries and with a focus on current and forthcoming regulatory requirements concerning the quality control and risk management for medical equipment. Q-Pro comprises a generalized application, providing the necessary flexibility to accommodate the different degrees of difficulty and specialization in creating or customizing QC protocols, carrying out inspections and managing collected data. The system incorporates a tool library for QC protocol design, widely used multimedia as well as a local database for protocol and inventory data archiving. The paper presents a detailed account of the system context of use, design and functionality. PMID- 11036579 TI - Platinum stimulating electrodes in physiological media. AB - The study reported here seeks to characterize behaviour of platinum electrodes of the 45-electrode spiral nerve cuff for selective electrical stimulation of different superficial regions of peripheral nerves in physiological solution (0.9% NaCl) and Eliott's buffered solution. Each electrode of the spiral cuff had a flat geometric surface of 2 mm2. To delineate an operational potential window between hydrogen and oxygen evolution during stimulation the electrochemical technique of cyclic voltammetry was used. In a typical cyclic voltammetry experiment, the potential of the tested electrode was cycled at an appropriate rate between two potential limits. The surfaces of the electrodes, obtained after injection of the biphasic charge as defined in a physiological solution (0.9% NaCl), thus simulating long-term electrical stimulation, were investigated using a high resolution Auger electron spectroscopy (AES) method. PMID- 11036580 TI - An overview of telemedicine: the virtual gaze of health care in the next century. AB - This article considers the role of telemedicine in the production of biomedical health care using three specific theoretical constructs as lenses through which to examine this phenomenon: (1) Foucault's medical "gaze"; (2) the political economy of health; and (3) deterritorialization and multisite ethnography. This examination focuses first on the changing corporate structure of health care and changing political attitudes toward telemedicine. Second, it documents the current use of telemedicine in prisons, the military, and in cross-cultural settings. Third, it discusses responses to telemedicine of individual physicians, health care staff, and patients, finding that these responses are broadly conditioned by an individual's mode of articulation with biomedical institutions and that they are mediated by personal experience. PMID- 11036582 TI - Medicalizing homelessness: the production of self-blame and self-governing within homeless shelters. AB - This article draws upon three years of ethnographic research within an emergency homeless shelter in Massachusetts to explore the subject-making effects of routine shelter helping practices. A medicalized discourse of deviancy is uncovered that provides the dominant conceptual framework within which both concerned homeless people and shelter staff remain enmeshed. As a result, helping practices focus on detecting, diagnosing, and treating understood deviancy within the bodies or selves of homeless people. The dominant discursive practices produce homeless subjects who learn to look within their selves for the "cause" of their homelessness. Treatment focuses on reforming and governing the self. Alternative discourses suggesting the need for practices challenging broader political economic processes are thus marginalized as peripheral and unreasonable. PMID- 11036581 TI - Witnessing and the medical gaze: how medical students learn to see at a free clinic for the homeless. AB - This article analyzes doctor-patient communication as it is taught to medical students in a student-run free clinic for the homeless. Moving beyond Foucault's concept of the medical gaze, it incorporates Byron Good's theorizing about the soteriological aspects of medicine and medical education as well as aspects of practice theory as illuminated by Anthony Giddens. Ethnographic examples illustrate the necessary tension between objectification and subject-making that exists in the specific practices engaged in by both students and preceptors at the clinic site. PMID- 11036583 TI - Taxonomic anxieties: Axis I and Axis II in prison. AB - This essay describes the use of the Diagnostic and Statistical Manual of Psychiatry in the prison setting. The distinction between the Axis I disorders (major mental illness) and those designated Axis II (character disorders) is explored in terms of the professional division of labor in prison and the problems posed for prison discipline by "behaviorally disturbed" inmates. Conflicts over diagnosis are placed in ethnographic and historical perspective and form the basis for a discussion of the problematic relationship between disciplinary space and issues of subjection and agency. PMID- 11036584 TI - Secret encounters: black men, bisexuality, and AIDS in Alabama. AB - Black men suffer the highest rates of HIV infection in Alabama. However, little is known about the HIV risks of this sector of the population, primarily because the current public health focus is on women and children. The dearth of research on HIV risk among black men in Alabama is addressed by drawing on focus group, elicitation, and key informant data from an ongoing epidemiologic study on AIDS in that state. These hypothesis-generating qualitative interviews were used to identify three high-risk scenarios: "sex for money or drugs"; "prison sex"; and "sneaky sex" by married or nominally heterosexual men. It was found that covert and unprotected sex among bisexually active black men was commonplace for reasons that included prostitution, habituation to same-sex relations during incarceration, and the desire to maintain a facade of heterosexuality in homophobic communities. It was concluded that bisexual activity is highly correlated with secrecy and unprotected sex. The risks of bisexuality among black men are exacerbated by incarceration, homophobia, drug use, and the prison and public health focus on surveillance rather than prevention. PMID- 11036586 TI - Chasing the dragon: the cultural metamorphosis of opium in the United States, 1825-1935. AB - Many things to many people, opium has played a role in the emergence of several power bases in the United States. In turn, these bases of power have shaped what opium is for the rest of us. Allopathic medicine brought opium and its derivatives under its control around the turn of the century, promulgating "addiction theory" and addiction clinics as part of its rise to preeminence among rival forms of medicine. Opium also played a role in the U.S.'s international economic and imperialistic ascendance. When politicians began to deploy a new discourse on opium early in this century, they were able to appropriate medical rhetoric. As the politics of opium heated up, some doctors were able to exploit the emerging politically inspired discourse to generate a subtly different medical knowledge of opiates and addiction while establishing a new subdiscipline with the political support of lawmakers and state institutions. PMID- 11036585 TI - Negotiating cultural consensus in a breast cancer self-help group. AB - This article describes a shared model of the breast cancer experience negotiated by the members of a spontaneously organized breast cancer self-help group in eastern North Carolina. In the course of sharing their personal experience narratives with one another, these women worked to negotiate points of agreement among the varying sources of knowledge and oftentimes conflicting belief systems they held about breast cancer. The synthetic model they created rejected many of the assumptions underlying the dominant biomedical view of cancer "survivorship," particularly its emphasis on the autonomous individual as decision maker and its attendant male-gendered sports and military imagery--assumptions that often implicitly structured the agendas and topics discussed in the formal, medically sanctioned support groups these women found unappealing. The implications for theories about the construction of shared cultural models and for continuing efforts to design support groups to meet the needs of a diverse patient population are explored. PMID- 11036587 TI - Purification and biochemical characterization of TC10. PMID- 11036588 TI - Expression and purification of Rho/RhoGDI complexes. PMID- 11036589 TI - Bacterial expressed DH and DH/PH domains. PMID- 11036590 TI - Biochemical analysis of regulation of Vav, a guanine-nucleotide exchange factor for Rho family of GTPases. PMID- 11036591 TI - Activation of Rac1 by human Tiam1. PMID- 11036592 TI - Activation of Rho GEF activity by G alpha 13. PMID- 11036593 TI - Purification and characterization of guanine nucleotide dissociation stimulator protein. PMID- 11036594 TI - Purification and biochemical activity of Salmonella exchange factor SopE. PMID- 11036595 TI - Stimulation of Rho GDI release by ERM proteins. PMID- 11036596 TI - Expression and activity of human prenylcysteine-directed carboxyl methyltransferase. PMID- 11036597 TI - Purification and evaluation of large clostridial cytotoxins that inhibit small GTPases of Rho and Ras subfamilies. PMID- 11036598 TI - Rho GTPase-activating toxins: cytotoxic necrotizing factors and dermonecrotic toxin. PMID- 11036599 TI - Measurement of GTPase.effector affinities. PMID- 11036600 TI - Purification and in vitro activity of Rho-associated kinase. PMID- 11036601 TI - Purification and in vitro activities of p21-activated kinases. PMID- 11036602 TI - Stimulation of phospholipase C-beta 2 by Rho GTPases. PMID- 11036603 TI - Regulation of phospholipase D1 activity by Rho GTPases. PMID- 11036604 TI - In vitro interaction of phosphoinositide-4-phosphate 5-kinases with Rac. PMID- 11036605 TI - Activity and regulation of p35/Cdk5 kinase complex. PMID- 11036606 TI - Actin assembly mediated by Arp2/3 complex and WASP family proteins. PMID- 11036607 TI - In vitro actin polymerization using polymorphonuclear leukocyte extracts. PMID- 11036608 TI - Determination of GTP loading on Rac and Cdc42 in platelets and fibroblasts. PMID- 11036609 TI - Determination of GTP loading on Rho. PMID- 11036610 TI - Use and properties of ROCK-specific inhibitor Y-27632. PMID- 11036611 TI - Inducible membrane recruitment of small GTP-binding proteins by rapamycin-based system in living cells. PMID- 11036612 TI - Expression of Rho GTPases using retroviral vectors. PMID- 11036613 TI - Expression of Rho GTPases using adenovirus vectors. PMID- 11036614 TI - In vivo activity of wild-type and mutant PAKs. PMID- 11036615 TI - Single cell assays for Rac activity. PMID- 11036616 TI - Effect of Rho GTPases on Na-H exchanger in mammalian cells. PMID- 11036617 TI - Actin filament assembly in permeabilized platelets. PMID- 11036619 TI - Isolation and in vitro contraction of stress fibers. PMID- 11036618 TI - Rho GTPases: secretion and actin dynamics in permeabilized mast cells. PMID- 11036620 TI - Inhibition of Rho GTPases using protein geranylgeranyltransferase I inhibitors. PMID- 11036621 TI - Imaging spatiotemporal dynamics of Rac activation in vivo with FLAIR. PMID- 11036622 TI - Rho-like GTPases in tumor cell invasion. PMID- 11036623 TI - Mouse embryo fibroblasts: a genetic model system for studying Rho- and Ras dependent cell cycle progression. PMID- 11036624 TI - Analyses of transforming activity of Rho family activators. PMID- 11036625 TI - Rho GTPases and cell migration-fibroblast wound healing. PMID- 11036626 TI - Rho GTPases and cell migration: measurement of macrophage chemotaxis. PMID- 11036627 TI - Rho GTPases and macrophage phagocytosis. PMID- 11036628 TI - Rho GTPases and axonal growth cone collapse. PMID- 11036629 TI - Study of in situ function of cytoskeletal proteins in lamellipodia and filopodia using microscale chromophore-assisted laser inactivation. PMID- 11036630 TI - Modulation of actin cytoskeleton by Salmonella GTPase activating protein SptP. PMID- 11036631 TI - The all purpose gene fusion. PMID- 11036632 TI - A practical guide to the construction and use of lac fusions in Escherichia coli. PMID- 11036633 TI - Tagging exported proteins using Escherichia coli alkaline phosphatase gene fusions. PMID- 11036634 TI - Applications of gene fusions to green fluorescent protein and flow cytometry to the study of bacterial gene expression in host cells. PMID- 11036635 TI - IVET and RIVET: use of gene fusions to identify bacterial virulence factors specifically induced in host tissues. AB - IVET was designed to identify those bacterial genes that are induced when a pathogen infects its host. A subset of these induced genes encode virulence factors, products specifically required for the infection process. The paradigm IVET system is based on complementation of an attenuating auxotrophic mutation by gene fusion and is designed to be of use in a wide variety of pathogenic organisms. In S. typhimurium, we have used this system successfully to identify a number of genes that are induced in a BALB/c mouse and that, when mutated, confer a virulence defect. The RIVET system is based on recombinase gene fusions, which, on induction during infection, mediate a site-specific recombination, the product of which can be screened for after recovery of bacteria from host tissues. In V. cholerae, we have used this system successfully to identify genes that are induced transcriptionally during infection of the gastrointestinal tract of infant mice. RIVET is also uniquely designed for postidentification analysis of in vivo-induced genes: (1) it has been used to analyze the temporal and spatial patterns of virulence gene induction during infection and (2) it has been used to dissect the regulatory requirements of in vivo induction with respect to both bacterial regulatory factors and host-inducing environments. The IVET system has several applications in the area of vaccine and antimicrobial drug development. This technique was designed for the identification of virulence factors and thus may lead to the discovery of new antigens useful as vaccine components. The IVET system facilitates the isolation of mutations in genes involved in virulence and, therefore, should aid in the construction of live-attenuated vaccines. In addition, the identification of promoters that are expressed optimally in animal tissues provides a means of establishing in vivo-regulated expression of heterologous antigens in live vaccines, an area that has been problematic previously. Finally, we expect that our methodology will uncover many biosynthetic, catabolic, and regulatory genes that are required for growth of microbes in animal tissues. The elucidation of these gene products should provide new targets for antimicrobial drug development. PMID- 11036636 TI - Identification of exported bacterial proteins via gene fusions to Yersinia pseudotuberculosis invasin. PMID- 11036637 TI - Use of imidazoleglycerolphosphate dehydratase (His3) as a biological reporter in yeast. PMID- 11036638 TI - Use of fusions to human thymidine kinase as reporters of gene expression and protein stability in Saccharomyces cerevisiae. PMID- 11036639 TI - Use of fusions to thymidine kinase. PMID- 11036640 TI - In vivo analysis of lacZ fusion genes in transgenic Drosophila melanogaster. PMID- 11036641 TI - Utility of the secreted placental alkaline phosphatase reporter enzyme. PMID- 11036642 TI - Fusions to imidazopyrazinone-type luciferases and aequorin as reporters. PMID- 11036643 TI - Novel methods for chemiluminescent detection of reporter enzymes. AB - Chemiluminescent reporter gene assays provide highly sensitive, quantitative detection in simple, rapid assay formats for detection of reporter enzymes that are widely employed in gene expression studies. Chemiluminescent detection methodologies typically provide up to 100-1000x higher sensitivities than may be achieved with fluorescent or colorimetric enzyme substrates. The variety of chemiluminescent 1,2-dioxetane substrates available enable assay versatility, allowing optimization of assay formats with the available instrumentation, and are ideal for use in gene expression assays performed in both biomedical and pharmaceutical research. In addition, 1,2,-dioxetane chemistries can be multiplexed with luciferase detection reagents for dual detection of multiple enzymes in a single sample. These assays are amenable to automation with a broad range of instrumentation for high throughput compound screening. PMID- 11036644 TI - Fusions to chloramphenicol acetyltransferase as a reporter. PMID- 11036645 TI - Fusions to beta-lactamase as a reporter for gene expression in live mammalian cells. PMID- 11036646 TI - Purification of proteins using polyhistidine affinity tags. PMID- 11036647 TI - Generating fusions to glutathione S-transferase for protein studies. PMID- 11036648 TI - Use of the Strep-Tag and streptavidin for detection and purification of recombinant proteins. PMID- 11036649 TI - Streptavidin-containing chimeric proteins: design and production. PMID- 11036650 TI - Fusions to maltose-binding protein: control of folding and solubility in protein purification. PMID- 11036651 TI - Thioredoxin as a fusion partner for production of soluble recombinant proteins in Escherichia coli. PMID- 11036652 TI - Affinity purification of recombinant proteins fused to calmodulin or to calmodulin-binding peptides. PMID- 11036653 TI - The S.Tag fusion system for protein purification. PMID- 11036654 TI - Fusions to self-splicing inteins for protein purification. PMID- 11036655 TI - Fusion proteins containing cellulose-binding domains. PMID- 11036656 TI - Biotinylation of proteins in vivo and in vitro using small peptide tags. PMID- 11036657 TI - Biotinylation of proteins in vivo: a useful posttranslational modification for protein analysis. PMID- 11036658 TI - Methods for generating multivalent and bispecific antibody fragments. PMID- 11036659 TI - Design and use of phage display libraries for the selection of antibodies and enzymes. PMID- 11036660 TI - Use of an Lpp-OmpA fusion vehicle for bacterial surface display. PMID- 11036661 TI - Identification of bacterial class I accessible proteins by disseminated insertion of class I epitopes. PMID- 11036662 TI - Bordetella pertussis adenylate cyclase toxin: a vehicle to deliver CD8-positive T cell epitopes into antigen-presenting cells. PMID- 11036663 TI - Use of anthrax toxin fusions to stimulate immune responses. PMID- 11036664 TI - Use of fusions to viral coat proteins as antigenic carriers for vaccine development. PMID- 11036665 TI - [On the 60th anniversary of the Department of Embryology of Moscow State University]. AB - The Department of Embryology of Moscow State University is 60 years old on September 1, 2000. This is not all that old, especially in comparison with the almost 250-year history of the university itself or the 150-year history of embryology in Russia, but it is sufficient, in looking back, to grasp our present situation in the common flow of events and time. PMID- 11036666 TI - [Work of morphogenesis in the Department of Embryology of Moscow State University: toward understanding the dynamic architecture of development]. AB - This is a review of studies on morphogenesis carried out at the Department of Embryology, Moscow State University, over the past 30 years. The main direction of studies has been to reveal and describe the properties of self-organizing fields of mechanical stresses in developing embryos. PMID- 11036667 TI - [The loach--the object of experimental embryologic research in the Department of Embryology]. AB - This paper provides a history of research at the Department of Embryology on the teleostean loach fish Misgurnus fossilis L. as a model. The use of the loach in education is briefly described. PMID- 11036668 TI - [Distant wave interaction in the early embryogenesis of the loach Misgurnus fossilis L]. AB - Groups of loach (Misgurnus fossilis L.) embryos of different ages were kept in different quartz cuvettes for 20-24 h so that only optic contact between the groups was possible. Subsequent observations showed that parameters of their development deviated from those in the control groups. Wave-mediated biocorrection proved to have both positive and negative effects, depending on the developmental stages of the interacting groups. Changes in spectral characteristics and polarization of biological radiation affected the results of the experiments. Various developmental abnormalities caused by distant wave mediated interactions of embryos and specific to each combination of developmental stages and conditions of optic communication are described. PMID- 11036669 TI - [Dynamics of the intensity of respiration in early embryogenesis of amphibians]. AB - We studied growth and respiration rate during early ontogenesis of the axolotl Ambystoma mexicanum, Bosca's newt Triturus waltlii, the green toad Bufo viridis, and the smooth clawed frog Xenopus laevis. The respiration rate in these amphibian species increases during embryonal and larval development, peaks after transition to active feeding, and decreases at later stages of ontogenesis. The patterns of dynamics of this energy metabolism index in tailed and tailless amphibians have some differences related to their specific development. The changes in respiration rate in the embryos and larvae are correlated with the concentrations of mitochondria. PMID- 11036670 TI - [Early embryo orientation by direction of the cardinal point in natural clutches of two ranidae species inhabiting geographically distant regions]. AB - The azimuth (the least angle with the north-south direction) of the first cleavage furrow and anteroposterior axes of neurula was measured on projections of photographs of natural clutches. The azimuth distribution of the craniocaudal axis of Rana dalmatina neurulae in clutches from southern Italy and the first cleavage furrows of R. arvalis embryos in clutches from central Russia proved identical. Both craniocaudal axes and first cleavage furrows were mostly oriented from west to east. The azimuth distribution of the craniocaudal axis of R. arvalis neurulae in clutches subjected to repeated cold shock proved to be random. The causes and mechanisms of predominant orientation of the embryos in natural clutches of frogs are discussed. We propose that magnetic sensitivity is acquired by cytoskeleton elements, most likely microtubules, during reorganization in the course of normal development or due to experimental influences. PMID- 11036671 TI - [Use of chimeric mice for studying the effects of genomic imprinting]. AB - This is a review of the data of clonal analysis of developing tissues in parthenogenetic and androgenetic chimeric mice. The time and causes of death of the parthenogenetic and androgenetic cell clones in chimeras are considered. The data obtained suggest that the development of cell clones, derivatives of the mesoderm and endoderm, is determined by the expression of alleles of the imprinted loci of paternal chromosomes, while the formation of cell clones, derivatives of the ectoderm, depends on the expression of other imprinted loci of maternal chromosomes. The death of androgenetic and parthenogenetic (gynogenetic) mammalian embryos is due to the lack of the expression of certain imprinted loci of the maternal and paternal genome, respectively. PMID- 11036672 TI - [Study of adhesive proteins from neural tissues of the 8-day old chicken embryonic eye]. AB - Two groups of proteins were isolated from the retina and pigment epithelium of eight-day-old chick embryos. Experiments with suspension cultures of retinal cells demonstrated that only the retinal extracts and the fraction of its acidic proteins can stimulate cell aggregation in vitro. Analysis by the method of high performance liquid chromatography showed that fractions of acidic and basic retinal proteins, which markedly differ in their electric charge and biological activity, have similar composition. To study the effect of these proteins on the morphological and functional state of pigment epithelium in vitro, a new experimental model is proposed, with the posterior segment of the newt (Pleurodeles waltl) eye used as a test tissue. The fraction of basic proteins isolated from the chick embryonic pigment epithelium stabilized cell differentiation in the newt pigment epithelium. The analyzed proteins proved to be biologically active at extremely low doses, corresponding to 10(-12) M solutions. PMID- 11036673 TI - [Discrete probability of a model for information provision on early embryonic development]. AB - A percolation model of the diffuse redistribution of morphogenetic information in early regulative development is analyzed. It is demonstrated that the statistical average values of cell connectedness remaining below the percolation threshold of the spatial redistribution of developmental determinants do not provide for the formation of cell structures of the necessary size. The average number of cell interactions should exceed the percolation threshold, and, therefore, the carriers of morphogenetic information in early development can move over distances comparable with the size of the entire embryo. The assumption concerning the percolation mechanism of cell death is used as a basis for estimating the statistical average value of cell connectedness at which the predicted number of cells theoretically isolated from the flow of signal molecules corresponds to the observed frequencies of dying embryonic cells. The estimated average number of cell interactions significantly exceeds the threshold of information resource percolation in the embryonic space and agrees with estimations of other authors, based on direct observations. The probable role of the diffusion front, or percolation cluster shell, in the regionalization of embryonic structures differing in their prospective values is discussed. It is shown that the duration of the communicative period, along with the statistical average number of channels providing for the intercellular transfer of signal molecules by diffusion, is a parameter controlling the processes of determination of embryonic structures. PMID- 11036674 TI - [Role of protein kinase A and calcium ions in regulating ovulation of oocytes by gonadotropins in the grass frog (Rana temporaria) in vitro]. AB - The inhibitor of cAMP-dependent protein kinase N-[2-(methylamino)ethyl]-5 isoquinolinesulfonamide (H-8) (12.5-50 microM) decreased the rate of ovulation of the follicle-enclosed Rana temporaria oocytes induced by the homologous pituitary extract in amphibian Ringer solution and in a chloride-free medium. The inhibitor of voltage-dependent calcium channels diltiazem (10 and 100 microM) decreased the rate of ovulation in Ringer solution but did not affect it in a chloride-free medium or decreased the ovulation inhibitory effect of this medium. It was concluded that cAMP-dependent protein kinase and intracellular free calcium ions were involved as second messengers in the gonadotropin regulation not only in maturation of amphibian oocytes but also in ovulation. PMID- 11036675 TI - [Activity of exogenous genetic designs, introduced into cells by a ballistic transfection method, in murine ontogenesis]. AB - We used the method of particle bombardment (ballistic transfection) to introduce beta-galactosidase and human dystrophin genes into mouse embryos and skeletal muscles of adult mice. We examined the mechanisms of DNA transfer into skeletal muscle cells, the biological processes accompanying and following this transfer, the susceptibility of various types of muscle cells to transfection, and the duration of expression of and conditions affecting the introduced genes. We have also developed an effective, convenient, and practical methods of skeletal muscles transfection. PMID- 11036676 TI - From polio to AIDS, 1950-2005. PMID- 11036677 TI - [What is the role of internal medicine in the collective of a general hospital?]. AB - General internal medicine tends to decrease in many Belgian hospitals. On the opposite, United States are now clearly engaged in a reform of their health system, with a drastic augmentation of the number of general internists. This reflection recognizes the specificities of general internal medicine: the first contact, the longitudinal follow-up, the comprehensive approach and the coordination with other medical participants. The need for subspecialists would be reduced and the need for primary comprehensive care physicians would be increased. The development of a performent sector of general internal medicine is able to respond to the demand of the patients and also of the general family practitioners. This model has positive impacts for the whole hospital community. We are all weary of discussions of state medicine, of the high cost of medical care, of the adequacy of medical care for the indigent, of the shortage of hospital beds for those who can pay, of the shortage of nurses, and so on. But these are, in part our problems; if their solution is to be to our liking, we must be active in them. The spector of state medicine is continually raised before us. Greater participation, by the federal, state and local governments in matters of health seems inevitable although most of us think it is important to retain in some manner or other the principle of private enterprise. Change of some sort will come; it is evident that unless we ourselves reorganize the practice of medicine, it will be reorganized for us. PMID- 11036678 TI - Serum lactate dehydrogenase activity increases in both endogenous and exogenous hypercorticisms. AB - Both endogenous and exogenous hypercorticisms are associated with a modest increase of the activity of serum lactate dehydrogenase. Considering both the wide use of this parameter and the frequent prescription of corticosteroids in clinical practice, awareness of this phenomenon is useful to avoid unnecessary investigations. PMID- 11036679 TI - The beta 3-adrenoceptor: physiological role and potential therapeutic applications. PMID- 11036680 TI - Repetitive analyses of P-glycoprotein in chronic myeloid leukaemia. AB - P-glycoprotein, a pump located in the plasma cell membrane, extrudes several clinically important drugs from the cell, and hence causes multidrug resistance. Reversing clinical drug resistance is possible by using agents that inhibit the activity of P-glycoprotein. We describe the results of sequential flow cytometric determinations of P-glycoprotein expression and activity in two patients suffering from acute lymphoblastic transformation of chronic myeloid leukaemia. Neither P-glycoprotein expression, nor its activity could be detected in the initial sample of the first patient. In the second patient, no P-glycoprotein expression was found at diagnosis. However, after chemotherapy containing P glycoprotein substrates, a significant expression was found in both patients and the functional flow cytometric test was positive. In order to achieve an accurate selection of patients that might benefit from the clinical use of P-gp inhibitors, repeated analyses are indicated in each patient suffering from acute leukaemia, during the course of the illness. PMID- 11036681 TI - Streptococcus milleri, a rare cause of pericarditis; successful treatment by pericardiocentesis combined with parenteral antibiotics. AB - We report the case of a 71-year-old woman suffering from purulent pericarditis, but displaying only minor symptoms. No predisposing factors were found. In addition, the infection was caused by Streptococcus milleri, a rare causative agent for this affection. We review the literature and discuss the different types of treatment of purulent pericarditis. PMID- 11036682 TI - Pasteurella dagmatis septicaemia in an immunocompromised patient without a history of dog or cat bites. AB - We report a rare case of Pasteurella dagmatis septicaemia in a 66-year-old immunocompromised patient, without a history of cat bites, dog bites or scratches. PMID- 11036683 TI - [Diagnosis and treatment of an unusual cause of metabolic acidosis: ethylene glycol poisoning]. PMID- 11036684 TI - Progress and prospects of ergot alkaloid research. AB - Ergot alkaloids, produced by the plant parasitic fungi Claviceps purpurea are important pharmaceuticals. The chemistry, biosynthesis, bioconversions, physiological controls, and biochemistry have been extensively reviewed by earlier authors. We present here the research done on the organic synthesis of the ergot alkaloids during the past two decades. Our aim is to apply this knowledge to the synthesis of novel synthons and thus obtain new molecules by directed biosynthesis. The synthesis of clavine alkaloids, lysergic acid derivatives, the use of tryptophan as the starting material, the chemistry of 1,3,4,5-tetrahydrobenzo[cd]indoles, and the structure activity relationships for ergot alkaloids have been discussed. Recent advances in the molecular biology and enzymology of the fungus are also mentioned. Application of oxygen vectors and mathematical modeling in the large scale production of the alkaloids are also discussed. Finally, the review gives an overview of the use of modern analytical methods such as capillary electrophoresis and two-dimensional fluorescence spectroscopy. PMID- 11036685 TI - Antimicrobial peptides of lactic acid bacteria: mode of action, genetics and biosynthesis. AB - A survey is given of the main classes of bacteriocins, produced by lactic acid bacteria: I. lantibiotics II. small heat-stable non-lanthionine containing membrane-active peptides and III. large heat-labile proteins. First, their mode of action is detailed, with emphasis on pore formation in the cytoplasmatic membrane. Subsequently, the molecular genetics of several classes of bacteriocins are described in detail, with special attention to nisin as the most prominent example of the lantibiotic-class. Of the small non-lanthionine bacteriocin class, the Lactococcus lactococcins, and the Lactobacillus sakacin A and plantaricin A bacteriocins are discussed. The principles and mechanisms of immunity and resistance towards bacteriocins are also briefly reported. The biosynthesis of bacteriocins is treated in depth with emphasis on response regulation, post translational modification, secretion and proteolytic activation of bacteriocin precursors. To conclude, the role of the leader peptides is outlined and a conceptual model for bacteriocin maturation is proposed. PMID- 11036686 TI - Biochemical engineering aspects of solid state bioprocessing. AB - Despite centuries of use and renewed interest over the last 20 years in solid state fermentation (SSF) technology, and despite its good potential for a range of products, there are currently relatively few large-scale commercial applications. This situation can be attributed to the complexity of the system: Macroscale and microscale heat and mass transfer limitations are intrinsic to the system, and it is only over the last decade or so that we have begun to understand them. This review presents the current state of understanding of biochemical engineering aspects of SSF processing, including not only the fermentation itself, but also the auxiliary steps of substrate and inoculum preparation and downstream processing and waste disposal. The fermentation step has received most research attention. Significant advances have been made over the last decade in understanding how the performance of SSF bioreactors can be controlled either by the intraparticle processes of enzyme and oxygen diffusion or by the macroscale heat transfer processes of conduction, convection, and evaporation. Mathematical modeling has played an important role in suggesting how SSF bioreactors should be designed and operated. However, these models have been developed on the basis of laboratory-scale data and there is an urgent need to test these models with data obtained in large-scale bioreactors. PMID- 11036687 TI - Multistage magnetic and electrophoretic extraction of cells, particles and macromolecules. AB - Improved techniques for separating cells, particles, and macromolecules (proteins) are increasingly important to biotechnology because separation is frequently the limiting factor for many biological processes. Manufacturers of new enzymes and pharmaceutical products require improved methods for recovering intact cells and intracellular products. Similarly isolation, purification, and concentration of many biomolecules produced in fermentation processes is extremely important. Often such downstream processing contributes a large portion of the product cost. In conventional methods like centrifugation and even modern methods like chromatography, scale-up problems are enormous, making them uneconomical and prohibitively expensive unless the product is of very high value. Therefore there has been a need for efficient and economical alternative approaches to bioseparation processes to eliminate, reduce, or facilitate solids handling. Magnetic and electric field assisted separations may hold considerable potential for providing a future major improvement in bioseparation technology. In the present review the merits and demerits of the existing methods are discussed. We present mainly our own research on the development of unified multistage extraction processes that are versatile enough to handle cells and particles as well as macromolecules as described below. We describe multistage methods, namely ADSEP (Advanced Separator), MAGSEP (Magnetic Separator), and ELECSEP (Electrophoretic Separator), for quantitatively separating cells, particles, and solutes by using magnetically and electrophoretically assisted extraction processes. To the best of our knowledge, multistage magnetic and electrophoretic separations have not been reported in the earlier literature. The theoretical underpinnings of these separations are crucial to their success and to the identification of their advantages over other separation processes in particular applications. Hence mathematical modeling is stressed here, presenting our own models while also reviewing models reported in the literature. We also present suggestions for future work while analyzing the scale-up and economic aspects of these extraction processes. Commercial uses of the magnetic and electrophoretic processes, having both ground- and space-based research elements, also are presented in this review. PMID- 11036688 TI - Recovery of proteins and microorganisms from cultivation media by foam flotation. AB - Foaming is often present in aerated bioreactors. It is undesired, because it removes the cells and the cultivation medium from the reactor and blocks the sterile filter. However, it can be used for the recovery of proteins and microorganisms from the cultivation medium. The present review deals with the characterization of model protein foams and foams of various cultivation media. The suppression of foaming by antifoam agents and their effect on the oxygen transfer rate, microbial cell growth and product formation are discussed. The influence of process variables on the recovery of proteins by flotation without and with surfactants and mathematical models for protein flotation are presented. The effect of cultivation conditions, flotation equipment and operational parameters on foam flotation of microorganisms is reviewed. Floatable and non floatable microorganisms are characterized by their surface envelope properties. A mathematical model for cell recovery by flotation is presented. Possible application areas of cell recovery by flotation are discussed. PMID- 11036689 TI - The natural functions of secondary metabolites. AB - Secondary metabolites, including antibiotics, are produced in nature and serve survival functions for the organisms producing them. The antibiotics are a heterogeneous group, the functions of some being related to and others being unrelated to their antimicrobial activities. Secondary metabolites serve: (i) as competitive weapons used against other bacteria, fungi, amoebae, plants, insects, and large animals; (ii) as metal transporting agents; (iii) as agents of symbiosis between microbes and plants, nematodes, insects, and higher animals; (iv) as sexual hormones; and (v) as differentiation effectors. Although antibiotics are not obligatory for sporulation, some secondary metabolites (including antibiotics) stimulate spore formation and inhibit or stimulate germination. Formation of secondary metabolites and spores are regulated by similar factors. This similarity could insure secondary metabolite production during sporulation. Thus the secondary metabolite can: (i) slow down germination of spores until a less competitive environment and more favorable conditions for growth exist; (ii) protect the dormant or initiated spore from consumption by amoebae; or (iii) cleanse the immediate environment of competing microorganisms during germination. PMID- 11036690 TI - Development of applied microbiology to modern biotechnology in Japan. AB - Development of modern biotechnology in Japan is characterized by unique contributions from applied microbiology and bioindustry. This review tries to summarize these original contributions with special emphasis on industrial production of useful substances by microorganisms. In the first part, development of applied microbiology and bioindustry in the last half of the twentieth century is summarized with a brief overview of the traditional background. In the second part, recent progress is reviewed with citation of typical achievements in biotechnology, applied enzymology, secondary metabolites, genetic engineering, and screening of microbial diversity, respectively. PMID- 11036691 TI - Microbial production of amino acids in Japan. AB - The microbial biotechnology of amino acids production which was developed and industrialized in Japan have been summarized. The amino acids include L-glutamic acid, L-lysine, L-threonine, L-aspartic acid, L-alanine, L-cysteine, L dihydroxyphenylalanine, D-p-hydroxyphenyl-glycine, and hydroxy-L-proline. PMID- 11036692 TI - Development of biotechnology in India. AB - India has embarked upon a very ambitious program in biotechnology with a view to harnessing its available human and unlimited biodiversity resources. It has mainly been a government sponsored effort with very little private industry participation in investment. The Department of Biotechnology (DBT) established under the Ministry of Science and Technology in 1986 was the major instrument of action to bring together most talents, material resources, and budgetary provisions. It began sponsoring research in molecular biology, agricultural and medical sciences, plant and animal tissue culture, biofertilizers and biopesticides, environment, human genetics, microbial technology, and bioprocess engineering, etc. The establishment of a number of world class bioscience research institutes and provision of large research grants to some existing universities helped in developing specialized centres of biotechnology. Besides DBT, the Department of Science & Technology (DST), also under the Ministry of S&T, sponsors research at universities working in the basic areas of life sciences. Ministry of Education's most pioneering effort was instrumental in the creation of Biochemical Engineering Research Centre at IIT Delhi with substantial assistance from the Swiss Federal Institute of Technology, Zurich, Switzerland to make available state-of-the-art infrastructure for education, training, and research in biochemical engineering and biotechnology in 1974. This initiative catalysed biotechnology training and research at many institutions a few years later. With a brief introduction, the major thrust areas of biotechnology development in India have been reviewed in this India Paper which include education and training, agricultural biotechnology, biofertilizers and biopesticides, tissue culture for tree and woody species, medicinal and aromatic plants, biodiversity conservation and environment, vaccine development, animal, aquaculture, seri and food biotechnology, microbial technology, industrial biotechnology, biochemical engineering and associated activities such as creation of biotechnology information system and national repositories. Current status of intellectual property rights has also been discussed. Contribution to the India's advances in biotechnology by the industry, excepting a limited few, has been far below expectations. The review concludes with some cautious notes. PMID- 11036693 TI - History of biotechnology in Austria. AB - Austria has contributed significantly to the progress of the biotechnologies in the past and is actively engaged in doing so today. This review describes the early history of biotechnology in Austria, beginning with the Vienna process of baker's yeast manufacture in 1846, up to the achievements of the 20th century, e.g. the submerged vinegar process, penicillin V, immune biotechnology, biomass as a renewable source of fermentation products (power alcohol, biogas, organic acids etc.), biopulping, biopolymers, biocatalysis, mammalian cell technology, nanotechnology of bacterial surface layers, and environmental biotechnology. PMID- 11036694 TI - Biotechnology in Hungary. PMID- 11036695 TI - Biotechnology in Switzerland and a glance at Germany. AB - The roots of biotechnology go back to classic fermentation processes, which starting from spontaneous reactions were developed by simple means. The discovery of antibiotics made contamination-free bioprocess engineering indispensable, which led to a further step in technology development. On-line analytics and the use of computers were the basis of automation and the increase in quality. On both sides of the Atlantic, molecular biology emerged at the same time, which gave genetic engineering in medicine, agriculture, industry and environment new opportunities. The story of this new advanced technology in Switzerland, with a quick glance at Germany, is followed back to the post-war years. The growth of research and teaching and the foundation of the European Federation of Biotechnology (EFB) are dealt with. The promising phase of the 1960s and 1970s soon had to give way to a restrictive policy of insecurity and anxiousness, which, today, manifests itself in the rather insignificant contributions of many European countries to the new sciences of genomics, proteomics and bioinformatics, as well as in the resistance to the use of transgenic agricultural crops and their products in foods. PMID- 11036696 TI - Independence of scientific publishing. Reaffirming the principle. AB - Historically, the American Psychological Association (APA), as part of its contract with the editors of APA journals, has granted to the editors sole responsibility to accept or reject manuscripts for publication, without such actions being open for review by APA. Such a policy fosters the freedom of inquiry and expression so necessary for a healthy science. It also serves to protect the Association because publication of an article in an APA journal does not represent endorsement by APA itself. The lessons of history are clear: When political or other pressures interfere with the autonomy of science, the societal and scientific consequences are grim. APA should reaffirm the principle that a healthy science of psychology requires an open exchange of ideas and findings. PMID- 11036697 TI - Will managed care change our way of being? AB - The authors studied psychotherapeutic practices commonly used in managed care settings and the theories and rhetorical strategies that justify them to speculate about if or how they are beginning to influence societywide understandings about the proper way of being human at the turn of the millennium. The practices--and effects--of managed care regulations on the self are interpreted by studying how the patient, the therapist, and the therapeutic relationship come to light in managed care settings. These categories are then used to speculate about the configuration of the newly emerging, 21st-century self. By extending hermeneutic concerns about instrumentalism and technicism, the authors suggest a new way of thinking about psychotherapy modeled less on positivist science and more on moral discourse. Finally, given this more hermeneutic understanding of psychotherapy, the authors speculate about alternative conceptions and arrangements of care. PMID- 11036698 TI - David and Goliath. When empirical and clinical standards of practice meet. AB - Traditional clinical methods of assessing the effectiveness of psychological treatments have come under attack. Experience and strong belief frequently lead to false confidence in treatments and sometimes result in damage to patients. Advocates have called for a scientific standard to replace the extant standards based on expert opinion and cost. Yet there are costs to the use of both the old standards and scientific standards based on manualized treatments and associated research. This article proposes a set of criteria for determining whether a treatment is scientifically credible based on empirically informed principles rather than on techniques or single-theory formulations. This proposal offers a way to overcome the problems of rigid manuals as well as those associated with forcing clinicians to adhere to theories and practices that are outside of their interest, experience, and expertise. Instead, scientifically sound, cross-cutting principles of treatment selection are proposed by which a treatment could be evaluated for scientific credibility and applied from a number of theoretical frameworks. PMID- 11036699 TI - And now what, after such tribulations? Memory and dislocation in the era of uprooting. AB - This article explores some of the often overlooked, traumatic psychological consequences that follow from major social and political disruption and upheaval. Specifically, it examines the importance of maintaining memory and legacy in the face of widespread uprootedness and dislocation of the sort that can undermine and even obliterate personal, social, and collective identities. The role that authentic social frameworks for memory play in preserving psychological rootedness and that accurate historical narratives play in resistance and regrouping is explored. Finally, the importance of creating a new psychology that is fully grounded in history and culture is emphasized. PMID- 11036700 TI - The scientific status of American psychology in 1900. AB - In 1900, psychologists were attempting to define themselves and searching for their role among both academic and non-academic public. The success of experimental methods served to advance their position as exemplary scientists, although, as the authors argue in this article, other factors were also important. First, the issue of measurement involved many disagreements about the tools needed to measure psychological constructs or even whether psychologists should measure anything at all. Second, the relationship between the brain and psychological constructs enhanced psychology's status for some, whereas others felt that psychologists should stay away from such topics. Parallels with present day concerns among psychologists are addressed at the end of the article. PMID- 11036701 TI - Philosophy of psychology at the turn of the century. AB - The philosophy of psychology at the turn of the century was an amalgam of perspectives and commitments--experimental science, Darwinian theory, positivism- forged partly out of achievements in experimental science and partly in response to transcendentalist (Hegelian) challenges. The amalgam itself appeared as an early version of the positivism that became developed and dominant early in the 20th century. For many psychologists at the turn of the century, experimental science, as practiced chiefly in physics and chemistry, was tantamount to a philosophy of science and one stripped of what were taken to be distracting and useless metaphysical quibbles. The assets and liabilities of this allegiance were recognized even as it was forming. PMID- 11036702 TI - A whisper of salvation. American psychologists and religion in the popular press, 1884-1908. AB - The advocates of the new psychology that emerged at the end of the 19th century were faced with a need to gain support from a public that was searching for a new basis for social and political order, yet was chary of any science identified with godless materialism. The first generation of American psychologists was faced with the dilemma of defining their approach as distinct from the old psychology while defusing public concern about the materialistic implications of their new science. Many of these new psychologists developed a rhetorical strategy of incorporating religious sentiment into their writing for the popular press. Their strategy emphasized the harmony of the new science with religious faith and stressed the moral qualities of psychological work. PMID- 11036703 TI - Snapshots of Freud in America, 1899-1999. AB - In 1899, Sigmund Freud was virtually unknown in America, and his new book. The Interpretation of Dreams, went unreviewed. A century later, his life and work were the subject of a major exhibition of the U.S. Library of Congress, and he appeared for the third time on the cover of Time magazine. This article describes a few of the intervening events. PMID- 11036704 TI - Psychotherapeutics and the problematic origins of clinical psychology in America. AB - The problematic place of psychotherapy within the larger history of scientific psychology is reviewed, especially in the absence of any definitive history of clinical psychology yet written. Although standard histories of psychology imply that psychotherapy was somehow derived from the tradition of German laboratory science, modern historiography reveals a dramatically different story. Personality, abnormal, social, and clinical psychology have their roots in an international psychotherapeutic alliance related more to French neurophysiology, and this alliance flourished for several decades before psychoanalysis. Reconstruction of the American contribution to this alliance, the so-called Boston school of abnormal psychology, suggests an era of medical psychology in advance of today. Note is also made of the possible misattribution of Lightner Witmer as the father of clinical psychology. PMID- 11036705 TI - Science, policy, and the protection of children. AB - Child maltreatment is a particular concern for many researchers, practitioners, and policymakers. The articles in this Child Sexual Abuse: Science, Practice, and Policy Issues section, edited by Jeffrey Haugaard, address this issue from several perspectives. Haugaard articulates the difficulties of defining certain types of abuse and of researching ambiguous variables. Karen Saywitz and her coauthors provide an overview of current treatments for child sexual abuse victims. Recent national statistics on child maltreatment and federal efforts to address the problem are described by Olivia Golden. Tom DeLay highlights current problems in the child welfare system and the federal government's role in responding to them. PMID- 11036706 TI - The challenge of defining child sexual abuse. AB - Although child sexual abuse has been a concern for many researchers, therapists, and advocates for the past 3 decades, several fundamental issues regarding child sexual abuse remain unresolved. In particular, the term child sexual abuse has never been unequivocally defined. The lack of a commonly accepted definition of child sexual abuse continues to inhibit research, treatment, and advocacy efforts. Early researchers used broad and inclusive definitions of child sexual abuse--definitions that often continue to be used today. The consequences of these definitions are discussed, and strategies for developing other definitions of child sexual abuse are suggested. PMID- 11036707 TI - Treatment for sexually abused children and adolescents. AB - The authors review research demonstrating the variable effects of childhood sexual abuse, the need for intervention, and the effectiveness of available treatment models. The well-controlled treatment-outcome studies reviewed do not focus on sensationalistic fringe treatments that treat sexually abused children as a special class of patients. Instead, studies demonstrate empirical evidence for extending and modifying treatment models from mainstream clinical child psychology to sexually abused children. The authors propose a continuum of interventions to meet the needs of this heterogeneous group. Interventions range from psychoeducation and screening, to short-term, abuse-focused cognitive behavioral therapy with family involvement, to more comprehensive long-term plans for multiproblem cases. Last discussed are gaps in the research and suggestions for future research to address the pressing dilemmas faced by clinicians and policymakers. PMID- 11036708 TI - The federal response to child abuse and neglect. AB - Addressing the problem of child maltreatment is a high priority for the Clinton administration. Guided by the principles of safety, permanency, and the child's well-being, the Administration on Children and Families (ACF) has made great strides in improving the lives of maltreated children. Critical programs administered by ACF include the Adoption and Safe Families Act, Community-Based Family Resource and Support Program grants, Children's Justice Act programs, and Child Abuse Prevention and Treatment Act research and demonstration projects. Projects serve both to expand existing programs and to develop innovative approaches. ACF has also sponsored several multidisciplinary national conferences designed to generate a sense of shared responsibility and a renewed commitment to solving problems of child abuse and neglect. PMID- 11036709 TI - Fighting for children. AB - Child abuse is a problem of national importance. Nationwide, billions of dollars are spent on the child welfare system, but child abuse rates remain high. At the federal level, new laws, such as the Child Abuse Prevention and Enforcement Act, the Foster Care Independence Act, and the Adoption and Safe Families Act, represent steps toward addressing the problem. However, increased efforts are needed to raise awareness of the problem of child abuse and to prevent it. Furthermore, more attention is needed to address the need for reform of the child welfare system and to involve the private sector in helping abused children. PMID- 11036710 TI - Incompatible with evolutionary theorizing. PMID- 11036711 TI - The Darwin is in the details. PMID- 11036712 TI - From allies to adversaries? PMID- 11036713 TI - Once again, the origins of sex differences. PMID- 11036714 TI - Formal logic and dialectical thinking are not incongruent. PMID- 11036715 TI - Dialectical thinking: neither eastern nor western. PMID- 11036716 TI - What is missing in Chinese-western dialectical reasoning? PMID- 11036717 TI - Dialectical responses to questions about dialectical thinking. PMID- 11036718 TI - Work to reduce stigma. PMID- 11036719 TI - Automated abrasive blasting equipment for use on steel structures. PMID- 11036720 TI - Ergonomics of rebar tying. PMID- 11036721 TI - Use of advanced training technologies by industrial hygienists: what can the ACGIH Computer Committee contribute? PMID- 11036722 TI - Respirable crystalline silica exposures during tuck pointing. PMID- 11036723 TI - 2000 Stokinger Lecture Award. Chemical exposure guidance levels consistency, integrity and public trust. PMID- 11036724 TI - The impact of recirculating industrial air on aircraft painting operations. PMID- 11036725 TI - Assessment of exposure to manganese in welding operations during the assembly of heavy excavation machinery accessories. AB - Welder exposure to metals in various industrial sectors is poorly characterized. We had the opportunity to carry out an exploratory study to characterize manganese exposure in welding operations in a recently established Quebec factory that assembled accessories for heavy excavation machinery. Ten workers were sampled for total manganese for at least two consecutive days out of three followed by two consecutive days for respirable manganese (with a size selective sampler with a median cut-off of 4 microns), during a typical week in the summer of 1998. Parts being welded were characterized as large or small. Small parts were those being welded on tables during subassembly. Workers were divided into two groups according to the parts they were welding. Seventy-eight percent of the total manganese exposure levels of welding operations during the assembly of large accessories of heavy excavation machinery exceeded the manganese American Conference of Governmental Industrial Hygienists (ACGIH) threshold limit value (TLV) of 0.20 mg/m3 (GM 0.24 mg/m3, n = 14) while none exceeded the TLV during the assembly of small pieces (GM 0.06 mg/m3, n = 8). Welding operations during the assembly of large heavy excavation machinery accessories may pose a significant health hazard. Considering the importance of task-related variables affecting exposure among workers, further studies are needed to better characterize exposure determinants of welding operations during the assembly of heavy excavation machinery accessories. PMID- 11036726 TI - Methods to lower the dust exposure of bag machine operators and bag stackers. AB - This article reviews various dust control technologies developed over the years at the Pittsburgh Research Laboratory of the National Institute for Occupational Safety and Health (NIOSH) to provide various options and alternatives to lower bag machine operators' and bag stackers' dust exposures. Dust exposure records for the past 20 years show that bag machine operators and bag stackers normally have the highest respirable dust exposures of workers at mineral processing plants. A substantial amount of research has been performed over the years to minimize the dust exposure to these workers and the intent is to present all this information together in one article. Most of the research describes engineering controls that were adapted to existing facilities to reduce the dust generated during bag filling, bag conveying, and bag stacking. In some cases, a single technique succeeded in lowering respirable dust concentrations for all three processes, thus reducing the dust exposure to both the bag machine operator and the bag stacker. In other cases, a technique was developed to specifically reduce the dust exposure of one process or the other. This research also reviews various controls for secondary dust exposure, including general ventilation requirements to mill buildings, the effects of background dust sources, and personal work practices. This information is presented to help industrial hygienists, plant managers, engineers, and workers lower the dust exposure of bag machine operators and bag stackers. PMID- 11036727 TI - Metal exposure among abrasive blasting workers at four U.S. Air Force facilities. AB - Button Aerosol Samplers were used to monitor the personal exposure of workers performing abrasive blasting operations at four U.S. Air Force facilities. Inhalable aerosols containing 25 metals, including cadmium, lead, and chromium, were investigated. The Button Aerosol Sampler was chosen because of its ability to successfully withstand mechanical stress, prevent very large particles from collection, and protect the filter from overloading and shredding by rebound particles. In addition, previous studies have shown that the sampling efficiency of this personal Aerosol Sampler exhibits low sensitivity to the ambient air conditions and that it adequately follows the inhalability convention. Inductively coupled plasma (ICP) was used to analyze the collected samples for all 25 metals. In addition, visual absorption spectrophotometry (VAS) was used to analyze for hexavalent chromium because of the presence of strontium chromate. The collected samples yielded 8-hr time-weighted average (TWA) concentrations that were up to 250, 6, and 5 times higher than the permissible exposure limits (PELs) for cadmium, lead, and hexavalent chromium, respectively. Also, the chromium levels measured by the ICP and VAS exceeded the strontium chromate threshold limit value (TLV) by up to 640 and 950 times, respectively. No correlation was found between the ICP and VAS hexavalent chromium concentrations. The likely reasons of this were the presence of Cr (II) and (III) that cannot be detected by the VAS, and the chemical interference from iron and some other metals in the samples. The Button Aerosol Sampler was shown to be useful for the monitoring of workers' exposure to heavy metals during abrasive blasting operations. PMID- 11036728 TI - Health effects of mycotoxins in indoor air: a critical review. AB - Industrial hygienists (IHs) are called upon to investigate exposures to mold in indoor environments, both residential and commercial. Because exposure standards for molds or mycotoxins do not exist, it is important for the industrial hygienist to have a broad knowledge of the potential for exposure and health effects associated with mold in the indoor environment. This review focuses on the toxic effects of molds associated with the production of mycotoxins, and the putative association between health effects due to mycotoxin exposure in the indoor environment. This article contains background information on molds and mycotoxins, and a brief summary and review of animal exposure studies, case reports, and epidemiological studies from the primary literature concerning inhalation of mycotoxins or potentially toxin-producing molds. The relevance of the findings in the reviewed articles to exposures to mold in indoor, non agricultural environments is discussed. Although evidence was found of a relationship between high levels of inhalation exposure or direct contact to mycotoxin-containing molds or mycotoxins, and demonstrable effects in animals and health effects in humans, the current literature does not provide compelling evidence that exposure at levels expected in most mold-contaminated indoor environments is likely to result in measurable health effects. Even though there is general agreement that active mold growth in indoor environments is unsanitary and must be corrected, the point at which mold contamination becomes a threat to health is unknown. Research and systematic field investigation are needed to provide an understanding of the health implications of mycotoxin exposures in indoor environments. PMID- 11036729 TI - Experimental evaluation of a mathematical model for predicting transfer efficiency of a high volume-low pressure air spray gun. AB - The transfer efficiency of a spray-painting gun is defined as the amount of coating applied to the workpiece divided by the amount sprayed. Characterizing this transfer process allows for accurate estimation of the overspray generation rate, which is important for determining a spray painter's exposure to airborne contaminants. This study presents an experimental evaluation of a mathematical model for predicting the transfer efficiency of a high volume-low pressure spray gun. The effects of gun-to-surface distance and nozzle pressure on the agreement between the transfer efficiency measurement and prediction were examined. Wind tunnel studies and non-volatile vacuum pump oil in place of commercial paint were used to determine transfer efficiency at nine gun-to-surface distances and four nozzle pressure levels. The mathematical model successfully predicts transfer efficiency within the uncertainty limits. The least squares regression between measured and predicted transfer efficiency has a slope of 0.83 and an intercept of 0.12 (R2 = 0.98). Two correction factors were determined to improve the mathematical model. At higher nozzle pressure settings, 6.5 psig and 5.5 psig, the correction factor is a function of both gun-to-surface distance and nozzle pressure level. At lower nozzle pressures, 4 psig and 2.75 psig, gun-to-surface distance slightly influences the correction factor, while nozzle pressure has no discernible effect. PMID- 11036730 TI - Work-related injuries in drywall installation. AB - Administrative data sources were used to describe the work-related injuries of drywall carpenters, to calculate rates of occurrence, and to explore high risk sub-groups. Health insurance eligibility files were used to identify a cohort of active union carpenters affiliated with a union local whose predominant work involved drywall installation in the state of Washington. These files contained the hours worked by each individual for each month between January 1989 and December 1995, providing person-hours at risk as a union carpenter. The Washington Department of Labor and Industries (L&I) provided records of workers' compensation claims filed by these individuals. Over seven years 1773 drywall carpenters filed 2567 workers' compensation claims representing an overall rate of 53.3 per 200,000 hours worked. These claims were filed by 1046 different individuals, or 59.0 percent of the cohort. Claims resulting in paid lost time from work were filed at a rate of 12.5 per 200,000 hours worked (n = 609) by 445 (25.1%) different individuals. The most common mechanisms of injury involved being struck (38.3%), overexertion (28.1%), and falls (13.2%). Struck by injuries most commonly involved cuts to the upper extremity. Overexertion injuries were most commonly described as sprains or strains involving the back. Sheetrock was associated with over 40 percent of these injuries. Falls most commonly involved injuries to the knee followed by the back and multiple injuries. Struck by injuries decreased steadily with increasing age and increasing time in the union. There was a steady increase in the rate of falls with increasing age. Overexertion injuries were responsible for the greatest proportion of costs for medical care, permanent impairment, and paid lost days. The high rates of overexertion injuries among these workers is consistent with known ergonomic stresses on drywall jobs. However, these workers are also at high risk of acute traumatic injuries. PMID- 11036731 TI - A new measure of parenting practices involving preadolescent- and adolescent-aged children. AB - Parenting behaviors have played critical roles in various theoretical and clinical models that have sought to explain the development and course of deviant child behavior. Notwithstanding the importance of this topic, the extant literature reveals very few adequate instruments for describing such patterns. To address 3 this need, the authors developed a self-report procedure for assessing parenting practices involving preadolescent and adolescent children, the Loeber Youth Questionnaire. Preliminary reliability and concurrent validity data were quite encouraging, and the instrument's two major dimensions (affect and control) fit well with the existing clinical and research literature. Limitations and future research directions are addressed. PMID- 11036732 TI - Treatment of GAD. Targeting intolerance of uncertainty in two types of worry. AB - This study evaluates the efficacy of a cognitive-behavioral treatment for generalized anxiety disorder (GAD) that addresses two types of worries: (a) those about situations that are amenable to problem solving, and (b) those about situations that are not. The treatment's goal is to help patients become more tolerant of uncertainty by discriminating between both types of worry and applying the correct strategy to each type. A multiple baseline design was used and subjects were 4 adults with a primary diagnosis of GAD. Treatment outcome was assessed with daily self-monitoring, self-report questionnaires, and standardized clinician ratings. At posttest and 6-month follow-up, 3 of 4 subjects no longer met diagnostic criteria for GAD and had attained high end-state functioning. At 12-month follow-up, none of the subjects met GAD diagnostic criteria but end state functioning was variable. The results also show that treatment outcome was highly related to change in intolerance of uncertainty. PMID- 11036733 TI - When instructions fail. The effects of stimulus control training on brain injury survivors' attending and reporting during hearing screenings. AB - Bedside hearing screenings are routinely conducted by speech and language pathologists for brain injury survivors during rehabilitation. Cognitive deficits resulting from brain injury, however, may interfere with obtaining estimates of auditory thresholds. Poor comprehension or attention deficits often compromise patient abilities to follow procedural instructions. This article describes the effects of jointly applying behavioral methods and psychophysical methods to improve two severely brain-injured survivors' attending and reporting on auditory test stimuli presentation. Treatment consisted of stimulus control training that involved differentially reinforcing responding in the presence and absence of an auditory test tone. Subsequent hearing screenings were conducted with novel auditory test tones and a common titration procedure. Results showed that prior stimulus control training improved attending and reporting such that hearing screenings were conducted and estimates of auditory thresholds were obtained. PMID- 11036734 TI - The Herald of Free Enterprise disaster. Lessons from the first 6 years. AB - A substantial body of data has been collected on survivors of the Herald of Free Enterprise disaster over the first 6 years. These data show the psychological effects to be considerable, and although they appear to decrease over time, 6 years later there remains a substantial minority that remains highly distressed. Our research has also pointed to those factors that appear to be important in determining the severity and chronicity of symptoms. Levels of crisis support early on seem to be protective. Not everyone has access to supportive others, and these people would seem to be at increased risk of disturbance. But even if crisis support is potentially available from family and friends, not everyone is in a position to draw on these resources. Those individuals who possess negative attitudes toward emotional expression might be less likely to seek out support. Evidence would suggest that modifying such attitudes might be an important component of intervention. A further target for intervention would seem to be the causal attributions made by survivors. It was found that those who perceived the causes of events during the disaster as internal and controllable were at greatest risk of psychological disturbance. The data gathered in the wake of this disaster suggest that intervening early with respect to these three components (crisis support, attitude to emotional expression, and attributional style) is highly likely to mitigate against long-term distress. PMID- 11036735 TI - Promoting independent task performance by persons with severe developmental disabilities through a new computer-aided system. AB - This study involved two experiments. In Experiment 1, a computer-aided system for promoting task performance by 6 persons with severe developmental disabilities was compared with a card system. The computer-aided system was portable and presented pictorial task instructions (one instruction per step) and prompts. In Experiment 2, the same system was used, but the number of instruction occasions was reduced. In one condition, the system presented all the instructions used in Experiment 1 but mostly in clusters rather than individually. In another, the system presented part of the Experiment 1 instructions. Three Experiment 1 participants also served in Experiment 2. Experiment 1 results indicated all 6 participants had higher percentages of correct steps with the computer system and preferred it to the card system. Experiment 2 results indicated that the condition in which the instructions were clustered was more effective for maintaining correct task performance. Implications of the findings were discussed. PMID- 11036736 TI - The relative effectiveness of EMDR versus relaxation training with battered women prisoners. AB - Five women prisoners with a history of being battered and who met the DSM-IV criteria for post-traumatic stress disorder were assessed (A phase) and provided with structured relaxation training (RT) (B phase, or placebo treatment), followed by eye movement desensitization and reprocessing (EMDR) therapy (C phase). Using the Beck Anxiety Inventory and the Impact of Events Scale's avoidance behavior and intrusive thoughts subscales as outcome measures, RT alone did not result in any clinical improvements. The subsequent provision of EMDR did not improve upon this lack of success with 4 of the 5 participants; 1 did improve on anxiety and intrusive thoughts. The apparent ineffectiveness of EMDR with these participants may be attributed to several explanations. Foremost perhaps is the hypothesis that EMDR is not sufficient to ameliorate the effects of chronic abuse. PMID- 11036737 TI - The development and validation of the attitude toward father scale. A tool for assessing the father's role in children's behavior problems. AB - Numerous measures assess how parental influences may relate to children's development of psychological difficulties. The majority of such measures focuses specifically on the mother-child relationship or assume both parents contribute equally and similarly to their children's psychological well-being. Previous research has largely ignored the need to assess the father-child relationship when examining parental influences on behavior problems. The goal of the present study was to develop and validate a self-report questionnaire to assess the father-child relationship. The Attitude Toward Father Scale was developed and validated using three independent samples of college undergraduates. The scale, which includes three subscales, was shown to have good psychometric properties. It was found effective in examining associations between father-child relationship scores and scores on adjustment measures. Findings showed total scale and subscale scores associated with measures of stress, alcoholism, hostility, depression, anxiety, and paranoia. Implications of findings and suggestions for future research are discussed. PMID- 11036738 TI - Higher education. PMID- 11036739 TI - Routine check-ups. PMID- 11036740 TI - Orthodontics on TV. PMID- 11036742 TI - Mixed messages. PMID- 11036741 TI - Teeth whitening. PMID- 11036743 TI - Special needs dentistry. PMID- 11036744 TI - Scleroderma presents as facial hypoaesthesia. PMID- 11036745 TI - Anti antibiotics? PMID- 11036746 TI - Specific clinical problem areas. AB - In this final article of the series, the clinical and technical aspects of a range of similar forms of immediate prosthesis are discussed. Similarly, copy denture and reline/rebase procedures are detailed. PMID- 11036747 TI - The treatment of adult patients with a mental disability. Part 3: The use of restraint. AB - Over recent years practitioners are increasingly being asked to attend to, or to provide treatment for, adult patients with some degree of mental illness, either in their homes or in the dental surgery. This final paper in the series deals with the lawful delivery of care or treatment in the face of resistance, through the use of restraint. PMID- 11036748 TI - Tobacco and oral disease. EU-Working Group on Tobacco and Oral Health. PMID- 11036749 TI - Ethnic and gender variations in university applicants to United Kingdom medical and dental schools. AB - AIM: To explore ethnic and gender variations amongst applicants to undergraduate United Kingdom medical and dental schools. METHOD: Retrospective analyses of University and College Admissions Services (UCAS) data on all students applying to study pre-clinical medicine and dentistry, during the academic years 1994/5, 1995/6 and 1996/7. Information for each medical and dental applicant included age, gender, social class and ethnic group. RESULTS: Of all applicants, just over half (50.2%) were male, though a greater proportion of applicants to dentistry were male (54.1%) than for medicine (49.3%) (OR = 1.21, 95% CI = 1.15, 1.28). Over one third (36.4%) of all students were from minority ethnic groups, a larger proportion of which were dental students (48.3%) than were medical students (33.8%) (OR = 1.83, 95% CI = 1.73, 1.94). There were also marked differences between medicine and dentistry when the ethnic groups were examined separately. The largest number of applicants from minority ethnic groups came from the Indian community, and this group increased in size annually by 4.1% (P < 0.05) for medicine, and 29% (P < 0.05) for dentistry. CONCLUSIONS: Significant inter-ethnic and gender differences are observed amongst applicants to medicine and dentistry. Dentistry appears to be relatively more attractive to minority ethnic applicants. PMID- 11036750 TI - Dental fluorosis in permanent incisor teeth in relation to water fluoridation, social deprivation and toothpaste use in infancy. AB - OBJECTIVES: To determine the prevalence and severity of fluorosis in permanent incisor teeth in young children in a fluoridated and a fluoride-deficient community and to establish what relationship, if any, there was between the occurrence of dental fluorosis and the reported use of fluoride toothpaste in childhood. DESIGN: A prevalence study of children aged 8-9 years who had been continuous residents in fluoridated Newcastle or fluoride-deficient Northumberland. METHOD: The permanent maxillary central incisor teeth were examined clinically and photographically by one examiner using the Thylstrup Fejerskov index; the photographs were read blind to child identity and clinical score. A closed-response questionnaire enquired into the child's early experiences of toothbrushing and use of fluoride toothpastes. Social deprivation was measured by a Jarman score. The study took place in 1998. OUTCOME MEASURE: Prevalence of dental fluorosis measured by the Thylstrup-Fejerskov index. RESULTS: Complete data were available for 78% (n = 409) and 79% (n = 403) of eligible sampled children in the two areas, respectively. Clinical and photographic results agreed closely and had high reproducibility. The prevalence of fluorosis was 54% in the fluoridated area and 23% in the fluoride-deficient area when all grades (> 0) of fluorosis were included; percentage prevalence of mild to moderate fluorosis (> or = 3) was 3% and 0.5% in the two areas, respectively. Multivariate analysis indicated that area of residence (odds ratio = 4.5), Jarman score (odds ratio = 0.99 per Jarman unit) and type of toothpaste (odds ratio = 1.6) were statistically significantly related to presence or absence of fluorosis: the risk factors were--fluoridated area, affluence, and use of adult toothpaste. CONCLUSIONS AND RECOMMENDATIONS: The prevalence of aesthetically important dental fluorosis was low, although higher in the fluoridated area. Use of a child's toothpaste (with lower fluoride concentration) could decrease risk in a fluoridated area. Adherence to the guidelines published by the British Society of Paediatric Dentistry is recommended. PMID- 11036751 TI - Scottish consortium for development and education in dental primary care. AB - A national consortium for dental primary care in Scotland has been formed as a result of integrated planning by groups involved in managing and delivering postgraduate dental education in Scotland. In 1998 a partnership was formed with representatives from the Scottish Council for Postgraduate Medical and Dental Education (SCPMDE) and the three Scottish dental institutions at Dundee, Edinburgh and Glasgow Universities. The principal aim of the Consortium is to promote evidence-based dental care through postgraduate education and research. The activities of the Consortium will provide a broadly based adjunct to current postgraduate provision with partner institutions contributing in different ways. Since its inception the Consortium has focused on two main areas: 'Higher Training' for Primary Care. Scottish Dental Practice Based Research Network. PMID- 11036752 TI - Human mesenchymal stem cells: progenitor cells for cartilage, bone, fat and stroma. PMID- 11036753 TI - Investigating the genetic control of stem cell behavior. AB - The precise control of hematopoietic stem cell fate decisions involves molecules that are differentially expressed in the stem cell versus its non-stem progeny. The construction of representative and high quality cDNA libraries containing stem cell-specific sequences is a first step in elucidating stem cell control mechanisms. Automated bioinformatics in conjunction with high-throughput random sequencing and high-density parallel array hybridization studies make it possible to dissect stem cell molecular pathways and networks. It is the differential analysis of these interacting pathways which will provide the greatest insights into the biological differences underlying stem cell behavior. PMID- 11036754 TI - Precursor B cells from Pax-5-deficient mice--stem cells for macrophages, granulocytes, osteoclasts, dendritic cells, natural killer cells, thymocytes and T cells. PMID- 11036755 TI - Insertional mutagenesis as a route to identifying genes involved in self renewal of haemopoietic stem cells. AB - The genes controlling self renewal in the haemopoietic system are still unknown. Using retroviral insertional mutagenesis we have established multipotent haemopoietic stem cell lines (FDCP-mix) that possess an increased self renewal capacity in vitro. To identify genes involved in the regulation of self renewal, proviral integration sites were cloned from FDCP-mix cells and used as probes to screen independently isolated FDCP-mix cell lines for a common proviral insertion site. So far, two common integration sites have been identified, A25 and M4. A25 is rearranged in 50% of the FDCP-mix cell lines and M4 in 10%. Genes located at or near these sites are likely candidates for the control of self renewal of haemopoietic stem cells. PMID- 11036756 TI - The helix-loop-helix inhibitor Id2 and cell differentiation control. PMID- 11036757 TI - Role of beta 1 integrin for hemato-lymphopoiesis in mouse development. PMID- 11036758 TI - The Ikaros family and the development of early intraembryonic hematopoietic stem cells. PMID- 11036759 TI - Function of cytokines in lymphocyte development. PMID- 11036760 TI - Re-evaluation of B lymphocyte lineage differentiation schemes. PMID- 11036761 TI - Growth factors regulating lymphatic vessels. PMID- 11036762 TI - Hemangioblastic precursors in the avian embryo. PMID- 11036763 TI - Cloning of JAM-2 and JAM-3: an emerging junctional adhesion molecular family? PMID- 11036764 TI - Thymus epithelial cell reaggregate grafts. PMID- 11036765 TI - A novel anti-Ep-CAM antibody to analyze the organization of thymic medulla in autoimmunity. PMID- 11036766 TI - Genetic dissection of thymus development. PMID- 11036767 TI - Developing thymocytes organize thymic microenvironments. PMID- 11036768 TI - The role of mesenchyme in thymus development. AB - We have reviewed the evidence that thymic mesenchymal cells and their progeny thymic fibroblasts play an important role in early T-cell development. Although it is possible that mesenchyme plays an inductive role in thymic epithelial morphogenesis, we have presented evidence to suggest that there is a direct effect of mesenchyme and fibroblasts on T-cell development. Moreover the association of these cell types with an ECM raises the possibility that the latter might be important in integrin and/or cytokine presentation especially during the CD4(-)8- phase of T-cell development. PMID- 11036769 TI - Making central T-cell tolerance efficient: thymic stromal cells sample distinct self-antigen pools. PMID- 11036770 TI - T cell activation and polarization by DC1 and DC2. PMID- 11036771 TI - ILT receptors at the interface between lymphoid and myeloid cells. PMID- 11036772 TI - Functional subsets of memory T cells identified by CCR7 expression. PMID- 11036773 TI - Functional organization of secondary lymphoid organs by the chemokine system. PMID- 11036774 TI - The cluster of ABCD chemokines which organizes T cell-dependent B cell responses. PMID- 11036776 TI - Precursors to neonatal lymph nodes: LT beta+CD45+CD4+CD3- cells are found in fetal liver. PMID- 11036775 TI - Affinity maturation in ectopic germinal centers. PMID- 11036777 TI - The role of tumor necrosis factor and lymphotoxin in lymphoid organ development. PMID- 11036778 TI - [Water requirement of food-producing animals and pets]. AB - In contrast to other essential nutrients there are only few publications dealing with the water requirement of food producing and companion animals. The exact derivation of water requirement and more detailed knowledge about the actual water consumption of each animal is only required if water supply is limited or too expensive to be provided in abundance. In case of limited water supply the water requirement is of special interest in order to prevent negative effects on animal health, performance or welfare. Intentions for water restriction or reasons for an accidental reduced water intake are quite different and variable in animal husbandry or keeping of companion animals. The following conditions only represent a few examples: water restriction in order to keep the litter dry (for example in poultry houses), due to technical problems concerning water supply like blocked-up waterers or failure to estimate the water requirement correctly (e.g. pet rabbits fed on concentrate and carrots without any additional water). Water consumption measured under conventional housing conditions (in litre per kg dry matter intake) varies in a wide range (from < 1:1 up to > 4:1) from one species to another. With higher performance leading to increased feed intake, water consumption usually also rises. For this reason it is very useful to calculate water consumption in relation to dry matter intake only. Besides there are several other factors of practical concern that have great influence on water intake: environmental temperature, intake of nutrients, that must be eliminated via the kidneys (e.g. electrolytes, nitrogen), as well as the amount of water spent for certain products like the sweat in horses or cow's milk. The importance of adequate water supply is best understood regarding the consequences of water deprivation (reduced feed intake, concentration of urine, impaired thermoregulation, reduced renal excretion of metabolic waste products, intake of other fluids that may be critical with regard to hygiene, behavioural problems). Because of the consequences of an inadequate water intake for animal welfare- last but not least--legislation on keeping animals provides guidelines for water supply that are controlled (among others) by veterinary administration. PMID- 11036779 TI - [Rules to control ddrinking water supply systems in livestock farming]. AB - Water is regarded as one of the most important distributors of infectious diseases. Moreover it can carry chemical compounds and toxic substances. Presently there are no specific legal regulations which define the quality of the drinking water for animals in Germany. However, some rules and criteria exist which help to survey and secure the water supply of animals on farm level. When controlling the hygienic quality of water on the farm it is necessary to investigate not only the water itself but also the local facilities around the water well. If health problems occur in the stock the water tanks, the tubing system which distributes the water in the animal houses and the drinkers have to be checked carefully. Water samples for the analysis on microorganisms and chemical contaminants should be taken according to a strict protocol. The regulations of the German drinking water directive for humans can be used as an orientation to characterise the drinking water quality for the animals. Nevertheless it seems useful to elaborate animal species specific thresholds for water contaminants particularly for food producing animals. A proposal for some threshold values is reported. PMID- 11036780 TI - [Microbial contamination in human and animal drinking water]. AB - Water plays an important roll in the epidemiology of a lot of infectious diseases. Groundwater as well as surface water contains generally microorganisms of several species, which cannot always differentiated properly in autochthonous flora and contaminants with health significance. Sources of bacterial or viral contaminants may be feces from man and/or animals but with different counts in ground and surface water. With respect to water used for supplying farm animals it is stated that it must have initially drinking water quality. Since it generally looses this quality in this supply system the following requirements shall be met: Free of Salmonellas and/or Campylobacter in 100 ml, no E. coli in 10 ml, total bacterial count at 37 degrees C less than 1000 cfu/ml and total bacterial count at 20 degrees C less than 10,000 cfu/ml. PMID- 11036781 TI - [Parasitic zoonotic disease agents in human and animal drinking water]. AB - Human- and veterinary important parasites of the subkingdom of protozoans and helminths infect humans and animals by ingestion of parasites in contaminated water. The parasites are excreted from the body of infected humans, livestock, zoo animals, companion animals or wild animals in the feces. Recreational waters, agricultural practices and wild animals serve as vehicles of transmission of the parasites in the water supplies. The following topics are addressed: a) the life cycles of parasitic diseases-causing agents with proven or potential transmission via water b) the development and the current research status of the analytical techniques for the detection of parasitic diseases-causing agents from water c) the occurrence of Cryptosporidium and Giardia in surface water supplies and in treated water d) the possible water sources and transmission ways of the parasites into the water supplies e) the behaviour and the possibilities for the removal or elimination of the parasites by water treatment. PMID- 11036782 TI - [Water as a vector for infectious stages of parasites in livestock]. AB - Water is a key factor in the life cycle of many parasite species. The development of the free-living generation (oocysts, cysts, eggs, larvae) and the longevity of the resulting infectious stages are extremely dependent on the availability of water. Damp stables are principally associated with an increased risk of parasite infection. Infectious stages may accumulate in dirty watering equipment and in pools of water on pasture. Utilization of surface water, especially when contaminated with sewage or effluent from sewage plants, imposes an increased risk of parasite infection on farm animals. Flooding of pastures with water containing parasite stages can lead to extensive contamination. Rainfall, flooding or inadequate land disposal of manure may contaminate surface water with parasite stages of animal origin from adjacent agricultural areas. PMID- 11036783 TI - [Examination of drinking water used in livestock production. Microbiological and physico-chemical methods. Ready to use test kits in field experiments]. AB - Livestock health care service is very much involved and interested in surveillance of the drinking water as well. However, in order to examine the water immediately "on the fly", test kits have to be provided, which offer results comparable to these obtained in the laboratories according to official prescription. The German Army was confronted with a similar situation during the secently performed mission in crisis regions. At the early state of a mission usually laboratory equipment is not yet established. Therefore a set of test kits was compiled suitable for mobile microbiological examination of drinking water. This set was excessively examined comparison with reference methods. In conclusion it is shown, that the mobile set gains equal or even better results compared to those obtained according to legally prescribed standard procedures. PMID- 11036784 TI - [Geologic and anthropogenic contaminants in drinking water]. AB - A review is given on the quality of water samples, regularly sent to the LUFA Oldenburg for analysis. Most of the samples derive from reduced groundwater bodies, indicated by a high concentration of iron (Fe, median 3 mg/l) and a low concentration of nitrate (NO3-, median 3 mg/l). Samples from peat land regions often show high concentrations of ammonium (NH4-) and a high oxidability. Samples from near the coast or near a salt mine may contain high salt concentrations. The deviation of the results is usually very high; in individual cases any parameter is able to make the water unusable. Each component may spoil the water for animal consumption if its concentration is too high. A scheme of standard values is given with the 4 graduations safe/enhanced/dangerous/unusable. Kind and age of the animals have to be taken into account, when water is assessed for animal consumption. The intake with the fodder of some substances such as Fe or NO3- is many times higher than the intake with the water. PMID- 11036785 TI - [Substances with pharmacological effects including hormonally active substances in the environment: identification of tetracyclines in soil fertilized with animal slurry]. AB - Many drugs used in human medicine are detectable in surface waters from the low to the very low microgram/L concentration range. In drinking waters only some of these substances were detected, the concentrations are usually an order of magnitude below the concentrations found in surface waters. A risk assessment of long time effects caused by a permanent intake of these low concentrations of drug residues cannot be done at this time. Hormonally active substances in surface waters may present an ecotoxicological risk, there are many investigations currently under way to assess this problem. Our investigations show for the first time that residues of the commonly used veterinary drugs tetracycline and chlortetracycline can be detected in the surface of soil (0-40 cm) fertilized with animal slurry. The maximum concentrations found were 32.3 micrograms/kg and 26.4 micrograms/kg respectively. Leaching of these compounds into seeping water sampled at a depth of 80-140 cm could not be detected with the methods employed. The significance of the detected antibiotic residues in soil samples for the quality of food of animal origin or any ecotoxicological consequences needs further investigations. The knowledge about the concentrations of veterinary drug residues resulting from animal husbandry in the environment is the first step for such a risk assessment. PMID- 11036786 TI - [Effect of drinking water on animal health: toxicologic health risks]. AB - To avoid health risks in farm animals it is necessary to limit the intake of unwanted chemical compounds via air, feed or drinking water. The basis for this procedure are experimental results of Veterinary Toxicology, after which acceptable daily intakes can be estimated. A health risk is not present, if those limits are not exceeded. Risks may not only occur as an impairment of health itself but as well as a reduction of yield of food producing animals (body growth, production of meat and fat, fertility, milk and egg production). In addition the cumulation of unwanted compounds in tissues, which are used for the production of food of animal origin, is of great importance for the limitation of daily intakes of those environmental pollutants. Following the results of toxicological experiments in the target animals "Toxicological Drinking Water Standards for Animals" can be established. PMID- 11036787 TI - [Drinking water as a medium for drugs]. AB - If drug treatment is necessary in large herd sizes, the administration with feed or drinking water is preferred. The use of the drinking water as a vehicle for drugs may be accompanied by several problems. An important condition for a successful medication is a good water solubility of the administered compound. The solubility is influenced by the quality of the used drinking water. The stability of the dissolved drug and incompatibilities with other water ingredients are additional limiting factors. The success of the treatment is influenced also by the taste of the medicated water which may cause a reduced water consumption. The use of tap water instead of spring water may be helpful to improve the efficacy of drug treatment via drinking water. PMID- 11036788 TI - [Water pollution, self-purification and restoration possibilities]. AB - Organic pollution is still the most important anthropogenic stress factor influencing the usability of ground and surface water as drinking water for cattle. Organic pollution is decomposed in several distinct heterotrophic processes that cause oxygen deficit, and finally end in the production of ammonia, hydrogen sulphide or methane. Even after the oxygen balance being restored higher nutrient concentrations (eutrophication) will remain. Eutrophication itself often leads to secondary pollution processes that adversely affect oxygen availability. Toxins may be generated by certain bluegreen bacteria growing under highly eutrophic conditions. Due to the concentrations of organic substances bacterial growth is forced, sometimes including faecal indicators and pathogenic species originating from the effluents of most of the communal sewage treatment works. Another kind of man made pollution concerns certain toxic substances. Disregarding the effects of accidental spills, the industrial production and use of pesticides in agriculture as well as other industrial chemicals or heavy metals may cause concentrations beyond acceptable limits. The use of surface water for cattle may become a problem for this reason. Accidental spills must be reckoned with as well, so the technical equipment for providing alternative sources of drinking water in emergencies must be available. Whereas the use of groundwater is normally not affected by toxic substance or pathogenic bacteria, high concentrations of iron, humic acids or salination may affect the suitability of water for cattle. PMID- 11036789 TI - [Do we need regulation of drinking water for animals? Recommendations for the water supply of farm animals and pets]. AB - The role of water as an essential nutrient for the animals, their health, performance and welfare is often underestimated. Presently there exists no specific legal regulation which defines the water quality for animal consumption. Some local recommendations make reference to the German regulation for the drinking water of humans. This seems to be useful in respect to food producing animals at a first glance. However, significant practical difficulties can occur. Therefore it is necessary to include in a possible regulation for the drinking water of animals additional animal and management specific aspects such as the type of animal, the keeping system, the water supply system, the distribution systems and the drinkers which are used. It may also be useful to define animal specific thresholds for certain groups of compounds and contaminants. As a first step towards a possible regulation 12 recommendations are given to improve and standardise the water supply of farm and pet animals. PMID- 11036790 TI - Naturally occurring isocyano/isothiocyanato and related compounds. PMID- 11036791 TI - Sulfur-containing amides from Glycosmis species (Rutaceae). PMID- 11036792 TI - Adoption, permanent care and foster care: home-based care in and beyond the 1990s. AB - Home-based care in Australia has changed considerably in the past two decades. The majority of children in adoptive, permanent care and foster families are likely to have experienced abuse, neglect and multiple placements. The disruptive behaviours displayed by these children undermine the potential for attachment offered by their new families. While the needs of the child will be obvious, the challenge for paediatricians is to recognize the relative instability of these newly established families and the high levels of stress they experience. PMID- 11036793 TI - Serological survey of measles and rubella immunity in Sydney preschool children. AB - OBJECTIVES: To estimate the prevalence of serological evidence of immunity to measles and rubella in preschool children in central and southern Sydney (NSW, Australia) and the prevalence of immunity in children with either documented or parentally reported immunization. METHODS: Geographical cluster random sampling was used to select children aged between 18 and 60 months to participate in the present study. Standardized interviews obtained information on each child's reported (by parents) immunization status and documentary evidence of immunization was recorded from the Personal Health Record. Venous blood was collected, serum was separated and stored frozen until tested. Measles and rubella antibodies were measured using ELISA, with either immunofluorescence or haemagglutination inhibition being used to clarify equivocal results. The study was conducted from 1992 to 1994 in conjunction with surveys of blood lead concentrations, iron status and micronutrient status. RESULTS: Parents of 726 of 953 children identified between 9 and 60 months of age agreed to participate in the lead, immunization, iron status and micronutrient studies. Sufficient blood for antibody testing was obtained from 580 children, aged 18 to 62 months at the time of collection. Parents reported that 94.7% (95% confidence interval (CI) 92.7-96.5%) of children had received a measles-mumps or measles-mumps-rubella (MMR) immunization. General practitioners administered 72.8% of these immunizations. The prevalence of serological evidence of immunity to measles and rubella was 88.8% (95% CI 86.2-91.4%) and 91.9% (95% CI 89.6-94.2%). respectively. There was documented evidence of measles and rubella immunization for 88.4% (95% CI 85.7-91.2%) and 86.4% (95% CI 83.4-89.3%) of children, respectively. Of children with documented measles immunization, 91.6% (95% CI 89.2-94.0%) had detectable measles antibody. Of children with documented rubella immunization 97.2% (95% CI 95.8-98.6%) had detectable rubella antibody. CONCLUSIONS: Measles and rubella immunization rates in central and southern Sydney are relatively high and most of these immunizations are provided by the private sector. Immunity to rubella in children with documented rubella immunization is at the level that would be expected from seroconversion studies. Immunity to measles in children with documented measles immunization is slightly lower than expected from seroconversion studies, highlighting the need for the second MMR immunization in preschool children, as well as making near universal immunization imperative if this disease is to be eradicated. PMID- 11036794 TI - Outcome of respiratory syncytial virus infection and a cost-benefit analysis of prophylaxis. AB - OBJECTIVES: To determine outcome and length of stay (LOS) for infants younger than 2 years of age admitted to hospital for respiratory syncytial virus (RSV) infection and to perform a cost-benefit analysis of prophylaxis with RSV gamma globulin (Respigam; CSL Laboratories, Melbourne, Victoria, Australia) and monoclonal antibody (Synagis; Abbott Australasia, Kurnell, NSW, Australia) in the Australian context. METHODS: Retrospective outcome study using data from an Australian hospital and intensive care databases over a 30 month period encompassing three RSV seasons (1 May 1997-31 October 1999). RESULTS: The mortality for RSV infection was very low, being 0.29% of all patients admitted to hospital and 0% in infants without co-existing disease. Only 11.4% of infants required admission to the intensive care unit (ICU). Respiratory syncytial virus infection was not associated with prolonged hospitalization; patients managed in the general wards had a mean length of stay (LOS) of 4.13 days with a median of 3.0 days, while those requiring intensive care had a mean LOS of 9.8 days (including 5.1 days in the ICU). Use of RSV prophylaxis would be expected to reduce the requirement for hospital admission; however, the cost of prophylaxis is conservatively estimated to be between 7.2- and 65.3-fold the money saved in hospital care, depending on the weight of the patient and choice of drug. CONCLUSIONS: Respiratory syncytial virus is a major cause of morbidity, but not mortality, in infants. In infants requiring hospitalization for this disease, the LOS is relatively short and the mortality extremely low. Use of prophylactic agents, which reduce the probability of hospital admission but have no effect on mortality, cannot be justified in the Australian context on cost-benefit grounds. PMID- 11036795 TI - Neonatal outcome of gastroschisis and exomphalos: a 10-year review. AB - OBJECTIVE: To study neonatal outcomes associated with gastroschisis and exomphalos in a regional neonatal unit. METHODS: A retrospective (1988-97) data analysis to study the effect of the type of defect/surgery, mode/place of delivery and associated anomalies on time to start and reach full feeds, duration of total parental nutrition (TPN) support and total hospital stay. Exact bivariate test procedures were used for data analysis. RESULTS: Twenty-one cases of gastroschisis (17 inborn) and five cases (four inborn) of exomphalos were identified. Of these, 23.8% cases of gastroschisis and 60% of cases of exomphalos had associated gut anomalies. The survival rates for gastroschisis and exomphalos were 91 and 100%, respectively. The median time to start and reach full enteral feeds in outborn neonates was longer than in inborn neonates (9 vs 25 days, respectively, P = 0.01; and 16 vs 49 days, respectively, P = 0.01), as was the duration of TPN support (14 vs 42 days, respectively; P = 0.02). Neonates with gastroschisis had significant delays in starting and reaching full feeds compared with neonates with exomphalos (median 13 vs 4.5 days, respectively, P = 0.03; and 24 vs 8, respectively, P = 0.02) and they required prolonged support with TPN (median 23 vs 6 days, respectively; P= 0.01). Antenatal detection was significantly more frequent in inborn compared with outborn neonates (100 vs 67%, respectively; P = 0.03). The severity of associated gut anomalies and the delivery to surgery interval did not differ significantly to explain the increased morbidity in outborn neonates. Outcome was not significantly different after analysis by type of surgery and mode of delivery. CONCLUSIONS: Increased morbidity in outborn neonates may be related to factors such as temperature, care, hydration status, care of the defect and vascular compromise of prolapsed gut during prolonged transportation. PMID- 11036796 TI - Child injury mortality in New Zealand 1986-95. AB - OBJECTIVES: Injury has been described as 'the last major plague of the young'. We provide an epidemiological description of injury, as a leading cause of death in New Zealand, and identify options for prevention. METHODS: We identified all deaths due to any cause for the period 1986-95 from the national data and calculated the potential years of life lost for each death. For the same period, we identified all injury deaths for children aged 0-14 years. Causes of injury were examined in four age groups. RESULTS: In the population as a whole, injury was the fourth leading cause of death and the leading cause of potential years of life lost. Injury killed children at the rate of 16.8 per 100,000 person-years. The victims were predominantly male (62%) and 52% were under 5 years of age. In infancy (<1 year of age), suffocation was the leading cause of injury mortality. From 1 to 14 years of age, motor vehicle traffic incidents were the leading cause of mortality. CONCLUSIONS: Motor vehicle traffic incidents, drowning, suffocation and suicide stood out as areas with the greatest potential for reducing child injury mortality. A number of existing prevention strategies show promise (e.g. child restraints), others are inadequately implemented (e.g. swimming pool fencing) or are of unknown efficacy (e.g. government suicide prevention policies). Strategies to reduce infant suffocation and child non-traffic pedestrian deaths remain to be developed and tested. PMID- 11036797 TI - Quality of life of mothers and families caring for preterm infants requiring home oxygen therapy: a brief report. AB - OBJECTIVE: To investigate the impact on mothers and families of caring for a premature infant receiving home oxygen therapy (HOT). METHODS: Standard questionnaires were used to compare the functioning of mothers and families of 10 premature infants receiving HOT, 10 premature infants who were discharged from hospital on HOT but who no longer required it and 20 premature infants who had never required treatment with HOT. RESULTS: After adjustment for gestational age, chronological age, birthweight and place of residence (urban/rural), the care required by premature infants receiving HOT had a significantly greater impact on their families than the care of infants not receiving HOT. Mothers of premature infants receiving HOT reported significantly less vitality and more mental health problems than mothers of infants not receiving HOT. CONCLUSIONS: The use of HOT for premature infants may have a significant adverse impact on their mothers and families. PMID- 11036798 TI - Epstein-Barr virus associated with immune thrombocytopenic purpura in childhood: a retrospective study. AB - OBJECTIVE: Although Epstein-Barr virus (EBV) is known to cause immune thrombocytopenic purpura (ITP), the epidemiology of this pathogen in children with ITP is not known. In the present study, the clinicoepidemiology and laboratory characteristics of EBV-associated ITP in childhood were analysed retrospectively. METHODS: The study cohort consisted of 108 children in whom ITP was diagnosed between 1990 and 1998. Patients were divided into EBV or non-EBV groups according to their serological status at diagnosis. RESULTS: Thirty-five (32.4%) of 108 children had ITP associated with acute EBV infection. The clinical manifestations and laboratory data were similar in children with and without acute EBV. Responses to various modalities of therapy were analysed. The average time to achieve complete remission (platelet count > or =150 x 10(9)/L) in EBV and non-EBV groups was 26 and 16 days, respectively. CONCLUSIONS: The incidence of childhood ITP associated with acute EBV infection is relatively high in Taiwan. Patients with EBV-associated ITP tended to resolve more slowly than those without EBV infection. PMID- 11036799 TI - Hepatic glycogenosis: reversible hepatomegaly in type 1 diabetes. AB - OBJECTIVE: To describe the aetiology, clinical features and appropriate treatment for hepatic glycogenosis in poorly controlled type 1 diabetes. METHODS: A review of three adolescents with poor diabetes control, hepatomegaly and elevated serum liver transaminase concentrations. RESULTS: Symptoms included abdominal pain, anorexia, nausea and vomiting. All had tender hepatomegaly; two had splenomegaly. Liver biopsy was performed on two patients. Histology revealed hepatic glycogenosis in both; one also demonstrated macrovesicular steatosis. With improved glycaemic control, all three showed resolution of their symptoms, organomegaly and elevated serum liver transaminase concentrations. CONCLUSIONS: Insulin-reversible hepatic glycogenosis is the most common cause of hepatomegaly and raised serum liver transaminase concentrations in children and adolescents with type 1 diabetes. Having excluded other causes of hepatic dysfunction, a 4 week therapeutic trial of improved glycaemic control is recommended prior to more invasive investigations. PMID- 11036800 TI - X-linked agammaglobulinaemia and the underlying genetics in two kindreds. AB - OBJECTIVE: Molecular analysis of the Bruton's tyrosine kinase (Btk) gene in two unrelated families, with a combined total of seven boys, affected by X-linked agammaglobulinaemia (XLA). METHODS: Protein electrophoresis and western blotting were used for the examination of Btk protein synthesis in blood leucocytes. Isolation of the coding sequence of the Btk gene was performed by amplification using the reverse transcription-polymerase chain reaction (RT-PCR) technique. Sequence alterations were screened for by the single-stranded conformation polymorphism (SSCP) method and characterized by standard sequencing protocols. RESULTS: Western blotting revealed Btk protein to be absent in leucocytes of affected males from both families. A novel 3 b.p. deletion in exon 3 of the Btk gene was found to be responsible for the XLA phenotype in the affected proband in one family (kindred I). A diagnostic PCR assay was established to detect this mutation in other affected male siblings and carrier females. For the second family (kindred II), the coding sequence of the Btk gene and the promoter region were found to be normal. CONCLUSIONS: The present study has demonstrated genetic heterogeneity in the Btk gene in South African XLA patients and has identified a novel mutation in this gene in the largest of the affected kindreds. The gene mutation in the second kindred was undetermined and may be indicative of a defect in some other gene associated with Btk function or stability. Western blotting was found to be informative in establishing a deficiency of Btk protein in both probands and is recommended as a frontline procedure in the molecular diagnosis and work-up of XLA. PMID- 11036801 TI - Medium-term outcomes are comparable with short-term outcomes in children with attention deficit hyperactivity disorder treated with stimulant medication. AB - OBJECTIVE: To compare the short- and medium-term effects of psychostimulant medication in children with attention deficit hyperactivity disorder (ADHD). METHODS: Seventy-three children with ADHD participated in a double-blind crossover study of dextroamphetamine (DEX) and methylphenidate (MPH; results previously reported). At the completion of this study, subjects continued to take the preferred stimulant. Subjects were restudied 6-9 months later. The principal outcome measures were the Revised Conners' Parent and Teacher Rating Scales. RESULTS: Fifty-three families (73%) returned the follow-up surveys. At 6-9 months, mean T scores were still significantly lower than the mean at baseline for all factors of both the CPRS-R and CTRS-R (P < 0.01). There were no statistically significant differences between scores at 6-9 months and scores at the completion of the corresponding medication period in the crossover trial. CONCLUSIONS: After 6-9 months treatment with stimulant medication, ratings remained significantly better than at baseline. This suggests that the early benefits of stimulants are sustained for at least 6 months. PMID- 11036802 TI - Cardiac responses to mild hypoxic hypercapnia in newborn babies: no effect of sleep position. AB - OBJECTIVE: To evaluate whether cardiac responses to a level of hypoxic hypercapnia that may be observed in rebreathing studies are altered with infant sleep position. METHODOLOGY: Eighteen healthy term infants (< 5-days-old) were studied. Heart rate (HR) and HR variability were monitored during air breathing and during 3 min exposure to a mixture of 15% O2/3% CO2 in both the prone and supine positions. Power spectral analysis of HR was performed. RESULTS: Heart rate was the only measured variable to be significantly changed in response to 15% O2/3% CO2. Hypoxic hypercapnia elicited no significant responses in power spectral HR variables. There was no effect of sleeping position on any of the measured variables. CONCLUSIONS: There are no significant differences in cardiac responses to mild hypoxic hypercapnia between sleep positions and power spectral indices of the autonomic control of HR are not altered by sleep position in newborn babies. PMID- 11036803 TI - Breastfeeding failure in a longitudinal post-partum maternal nutrition study in Hong Kong. AB - OBJECTIVE: To describe factors associated with breastfeeding failure during the first 6 months post-partum in a sample of Hong Kong Chinese women participating in a longitudinal study of maternal nutrition. METHODOLOGY: Forty-four Hong Kong Chinese lactating mothers who intended to breastfeed exclusively for at least 3 months were recruited and followed for 6 months post-partum. Demographic data were compared with 20 mothers who intended to use formula feeding. Mothers were followed up at 2 and 6 weeks and 3 and 6 months and details of infant feeding practices were obtained. Information was sought on breastfeeding management in hospital, reasons for discontinuation of breastfeeding or for providing supplements to babies and intention to seek, and sources of, lactation support. RESULTS: Thirty-nine mothers who planned to breastfeed completed the follow up. Compared with mothers in the formula-feeding group, breastfeeding mothers were more likely to be professionals or housewives. Continuation of any breastfeeding (total and partial) was noted in 30 (77%), 22 (57%), 16 (41%) and 12 (31%) mothers at 2 and 6 weeks and 3 and 6 months post-partum, respectively. The majority (97%) of mothers stated that they were given information on the benefits and management of breastfeeding. However, late initiation of breastfeeding and providing supplements to babies were common. Perceptions of insufficient milk supply (44%), breast problems (31%) and being too tired (28%) were the main reasons stated for stopping breastfeeding or for providing supplements to babies. Midwives from the postnatal wards and hotlines were the main sources of lactation support. CONCLUSIONS: These results highlight difficulties in sustaining breastfeeding, either exclusive or partial, in Hong Kong Chinese women. Despite being recruited on the basis of intending to exclusively breastfeed for 3 months, less than half these mothers were still breastfeeding and only approximately one third were exclusively or predominantly breastfeeding at 3 months. More needs to be done within the hospital environment to initiate breastfeeding immediately after birth and to avoid giving unnecessary supplements and more effort is needed to foster a mother's confidence, commitment and knowledge of breastfeeding. PMID- 11036804 TI - Gastroschisis: early enteral feeds may improve outcome. AB - OBJECTIVE: Population-based retrospective review of gastroschisis from 1986 to 1996. METHODS: This was a retrospective review of gastroschisis. Seventy cases were identified from the Birth Defects Registry of Western Australia (WA). Hospital medical records of live-born cases were reviewed. RESULTS: The live-born incidence of gastroschisis in WA was 2.1 per 10,000 live births for the period 1986-96. The incidence in mothers aged less than 20 years was 8.3-fold that of women aged over 30 years (P < 0.0001). The incidence rate for the period 1995-96 was over twice the rate for 1986-88. Age at first enteral feed was significantly related with length of hospital stay and duration of total parental nutrition (TPN). Each day delay in commencing enteral feed was associated with an increase in hospital stay of 1.05 days and an increase in TPN duration of 1.06 days. The method of delivery of the infant, age at repair, length of anaesthetic time, duration of postoperative paralysis and gestational age was not associated with length of stay or TPN duration. The data were divided into two cohorts: (i) 1986 90; and (ii) 1991-96. There was a statistically significant reduction in hospital stay from a geometric mean of 45.7 (1986-90) to 22.9 days (1991-96). CONCLUSIONS: Gastroschisis has a favourable outlook, with 89.7% survival of live births. Over the 10 year period studied, there has been a reduction in length of hospital stay and duration of TPN. The age at which the infant is first fed enteral feeds appears to be important in affecting the length of hospital stay and the duration of TPN, with delays associated with a longer hospital stay and longer TPN duration. PMID- 11036806 TI - Does maternal obesity adversely affect breastfeeding initiation and duration? AB - OBJECTIVE: To examine the relationship between maternal obesity and the initiation and duration of breastfeeding. METHODS: Analysis was made of the 1995 National Health Survey, in which personal interviews were conducted on a multistage area sample of private dwellings and a list sample of non-private dwellings in all states and territories of Australia. Mothers between the ages of 17 and 50 years (n = 1991) with children under the age of 4 years in 1995 participated in the study. RESULTS: Of the group of mothers with a body mass index (BMI) of 20-25, 89.2% (95% confidence interval (CI) 87.4-91.0) initiated breastfeeding, compared with 82.3% (95% CI 77.6-87.0) of mothers with a BMI of 30 or more. There was also a significant difference between the mean and median duration of breastfeeding of obese and non-obese mothers (BMI 30 and over, < 25, respectively). These differences remained significant when maternal smoking, age and other sociodemographic factors were taken into consideration. CONCLUSIONS: Health professionals should be aware that obese women may be at increased risk of not breastfeeding or stopping breastfeeding prematurely. PMID- 11036805 TI - Regional differences in outcome for very low-birthweight infants: do they persist at 7-8 years of age? AB - OBJECTIVE: To determine whether regional differences in early neonatal morbidity in a national cohort of very low-birthweight (VLBW) infants persisted at 7-8 years of age. METHODS: Perinatal data collected prospectively from birth on all VLBW infants born in New Zealand in 1986 and admitted to a neonatal unit included the hospital principally caring for the infant: hospitals A-D being level III hospitals and 'Other' including the smallest level III and all level II hospitals. At 7-8 years of age, 298 surviving children (96% survivors living in New Zealand) were assessed at a home visit. Parents were given a comprehensive questionnaire to complete, the children underwent a visual examination and were tested with the Revised Wechsler Intelligence Scale for Children and the child's teacher was sent a questionnaire to complete. RESULTS: Neonatal survival was significantly greater in the two largest hospitals (A and B) and this difference in survival remained at 7-8 years of age after adjustment for perinatal factors (P < 0.05). There were no differences between hospitals in risks of long-term sensorineural disability and behavioural or educational outcomes. There were interhospital differences in rates of visual problems and, after adjustment for confounding factors, there remained a marginally significant (P = 0.06) increased risk of myopia in hospital D. CONCLUSIONS: Despite differences in early morbidity favouring larger hospitals, there were no substantive differences in long-term (7 8 years) outcomes across a range of measures in this national cohort of VLBW infants. PMID- 11036807 TI - Ureaplasma urealyticum and its association with chronic lung disease in Asian neonates. AB - OBJECTIVE: The aim of the present prospective cohort study was to evaluate the relationship between lower respiratory tract colonization with Ureaplasma urealyticum and development of chronic lung disease (CLD) in a high-risk neonatal population. METHODS: Prospective cohort study of preterm infants with a birthweight < 1,500 g needing mechanical ventilation within 24 h of birth in a tertiary care neonatal unit. Endotracheal aspirates from these infants were cultured within 24 h for U. urealyticum and the rate of colonization was determined. The primary outcome measure was the incidence of CLD at 28 days of life. RESULTS: Of the 41 infants studied, 10 (24%) infants were colonized with U. urealyticum. The colonization rate was higher in babies < 1,000 g compared with babies weighing 1,000-1,500 g (P = 0.04). There was no significant difference between the colonized and non-colonized groups with regard to the antenatal use of steroids, maternal prolonged rupture of membranes, gestational age, birthweight, sex, respiratory distress syndrome, use of surfactant, patent ductus arteriosus and gastrooesophageal reflux. Of the 37 survivors, 20 (54%) developed CLD; eight infants (88.5%) in the colonized group developed CLD compared with 12 infants (42.8%) in the non-colonized group (P = 0.01). CONCLUSIONS: Neonates colonized with U. urealyticum were twice as likely to have CLD than non-colonized babies (relative risk 2.01; 95% confidence interval 1.27-3.37). These data suggest a significant association between colonization with U. urealyticum and CLD in infants weighing < 1,500 g. PMID- 11036808 TI - Implementation of evidence-based management of acute bronchiolitis. AB - OBJECTIVE: Implementation of clinical guidelines is frequently delayed well beyond their dissemination and the publication of clinical evidence. The recently published Australian guidelines for the management of acute viral bronchiolitis (AVB) have been evaluated by assessing the current practice of Australian paediatricians. METHODS: Questionnaire survey of all Australian paediatricians and a review of the literature. RESULTS: Of a total of 891 questionnaires, 555 (62%) were returned. Of the respondents, 373 (67%) treated children with AVB and, of these, 232 (67%) treated 10-50 children per year. A wide variation in management practice for both outpatient and inpatient treatment of AVB was identified. Up to 70% of paediatricians who treated AVB indicated using pharmaceutical agents in their outpatient management (88% in inpatient management), most using these agents 'sometimes' or in high-risk children. Paediatric respiratory physicians tended to use bronchodilators less frequently than general paediatricians. Compared with many countries in Europe, few Australian paediatricians routinely use supplementary drugs in the inpatient managenment of AVB; in particular, bronchodilators (61 vs 7%) and corticosteroids (11 vs 1%) are used far less often. A review of the literature demonstrated that pharmaceutical agents do not influence the course of AVB. CONCLUSIONS: Despite a lack of evidence for their efficacy and the recommendation of the Australian guidelines, pharmaceutical agents are frequently used in the management of AVB by paediatricians in Australia, although far less than reported in a recently published European survey. Guidelines alone are not sufficient to implement change and there is a need for more specific strategies to ensure that children receive appropriate management for this common condition. PMID- 11036809 TI - Errors in the certification of neonatal death. AB - OBJECTIVE: To examine the precision of the perinatal death certificate (PDC) and ascertain the possible sources of error in the certification of neonatal deaths. METHODS: The 'Main' and 'Other' causes of death recorded on the PDC were obtained from the Registry of Births, Deaths and Marriages and compared with those from a clinicopathological summary (CPS) completed after all pending laboratory results and/or autopsy information were available. RESULTS: There were 179 neonatal deaths during the 7 year period under review. The PDC and CPS main causes of death were concordant in 103 of 179 infants (58%) and discordant in the remaining 76 infants (42%). The PDC main cause of death was incorrectly classified in 61 of 76 infants (80%) with discordant findings and was incompletely classified in the remaining 15 infants (20%). The following discordancies were recorded for the 61 infants with an incorrect classification: (i) transposition of the 'Main' and 'Other' causes of death, resulting in a sequencing discordancy in 14 infants (23%); (ii) recording a non-pathological condition as the main cause of death in 40 infants (66%); and (iii) recording an incorrect pathological condition as the main cause of death in seven infants (11%). Eight of the 61 (13%) incorrect classifications and four of the 15 (27%) incomplete classifications were associated with laboratory and/or autopsy data being unavailable when the PDC was completed. CONCLUSIONS: The concordancy between the PDC and CPS would have increased from 58 to 91% if the 'Main' and 'Other' causes of death had been sequenced correctly, if the main cause of death had been ascribed to a pathological disease rather than a non-pathological condition and if corrective information from pending laboratory tests and/or autopsy examination had been made available to the Registry of Births, Deaths and Marriages. PMID- 11036810 TI - 3: Basic concepts of statistical reasoning: standard errors and confidence intervals. PMID- 11036811 TI - Is it Kawasaki disease? AB - Atypical cases of KD are common (up to 10% of the total) and the diagnosis should be considered without the full complement of diagnostic criteria. The risk of coronary dilation is high if IVIG is not given. Administration of IVIG is effective at preventing aneurysms, if given early. The high-risk groups for coronary artery disease are infants younger than 6 months of age and older children with very high platelet counts, high ESR and fever persisting for more than 2 weeks. PMID- 11036812 TI - Central nervous system candidiasis in preterm infants: limited value of biochemical markers for diagnosis. AB - Two rare cases of isolated central nervous system (CNS) candidiasis in preterm infants have been diagnosed in a tertiary neonatal centre over the past 6 years. Despite the life-threatening nature of the disease, biochemical infection markers were not useful for the early identification of localized fungal infection. Because the infection was likely to have been blood borne, we postulated that the initial fungal load was probably low and that the organisms were rapidly eliminated from the circulation after a few had been deposited in the CNS. Hence, the absence of fungaemia or systemic involvement precluded the activation of cytokines and cellular markers. Clinicians should be aware of the limitation of biochemical infection markers so that diagnosis and treatment of fungal infection will not be delayed. PMID- 11036813 TI - Tension pneumo-orbitus and pneumocephalus induced by a nasal oxygen cannula: report on two paediatric cases. AB - The present paper highlights the potential dangers of misplaced nasopharyngeal oxygen cannulae causing secondary pneumo-orbitus and pneumocephalus in two paediatric patients. While this complication is uncommon, early recognition allows prompt and appropriate intervention, with cessation of nasal oxygen, cannula removal, early investigation with computed tomography (CT) head/orbit scan and orbital or cranial decompression, if required. Early CT imaging identifies medial orbital or paranasal sinus fractures, the presence of sinusitis, associated intracranial air and assessment of the degree of orbital or intracranial tension. Antibiotics are not usually required for this type of clean injury unless pre-existing sinusitis is identified. In both cases, direct orbital decompression was performed with excellent results after identification of marked unilateral tense exophthalmos, delayed pupillary reactions to light and ophthalmopegia. PMID- 11036814 TI - Ataxia with isolated vitamin E deficiency: a clinical, biochemical and genetic diagnosis. AB - A case of ataxia with isolated vitamin E deficiency, in conjunction with supportive genetic studies, is reported. This is a neurodegenerative condition that involves a mutation in the tocopherol (alpha) transfer protein gene (TTPA). Measurement of serum vitamin E concentration should be included as part of the investigations in children with progressive ataxia, even in the absence of fat malabsorption. Early treatment with vitamin E may protect such patients against further neurological damage. PMID- 11036815 TI - Unilateral right-sided internal jugular phlebectasia in asthmatic children. AB - The most common cause of a neck mass that increases in size on straining is laryngocele. Internal jugular phlebectasia, which is of unknown cause, may present similarly. We present three cases of internal jugular phlebectasia, all of whom were asthmatic children. This association of asthma and internal jugular phlebectasia has not been reported previously. PMID- 11036816 TI - Infectious mononucleosis-like illness in an infant with measles. PMID- 11036817 TI - Automatic car locking and toddler entrapment. PMID- 11036818 TI - Parental awareness of side effects associated with the use of inhaled corticosteroids in Malaysian asthmatic children. PMID- 11036819 TI - Ankylosing spondylitis--at the interface of bone and cartilage. PMID- 11036820 TI - Enthesitis, osteitis, microbes, biomechanics, and immune reactivity in ankylosing spondylitis. PMID- 11036821 TI - Fish oil and rheumatoid arthritis: antiinflammatory and collateral health benefits. PMID- 11036822 TI - A DNA polymorphism at the alpha2-macroglobulin gene is associated with the severity of rheumatoid arthritis. AB - OBJECTIVE: To determine if DNA polymorphisms at the alpha2-macroglobulin (alpha2m) and angiotensin converting enzyme (ACE) genes were associated with rheumatoid arthritis (RA). METHODS: A total of 160 patients (71 with early active severe RA, 89 with non-severe RA) were genotyped (polymerase chain reaction) for the alpha2m (5 bp deletion/insertion) and ACE (I/D) polymorphisms. We also genotyped 500 healthy controls from the same Caucasian population (Asturias, Northern Spain). RESULTS: Carriers of the alpha2m deletion allele were at a significantly higher frequency among patients with an early active severe form of the disease, compared to patients with non-severe RA (p = 0.037). The frequency of the alpha2m deletion allele was significantly higher in patients with severe compared to nonsevere RA (p = 0.017). In addition, the frequency of the deletion allele was significantly higher among patients with 5 or more episodes of acute exacerbation of disease activity per year (n = 39) compared to those with none (n = 46) (p = 0.002). Gene and genotype frequencies for the ACE-I/D polymorphism did not differ between those with early active severe and non-severe RA. CONCLUSION: The genetic variation at alpha2m is associated with the severity of RA. Carriers of the alpha2m deletion allele would have increased risk of developing an early active severe form of the disease. Our data suggest that alpha2m could be a valuable target in the treatment of RA. PMID- 11036823 TI - Metabolic activation stimulates acid production in synovial fibroblasts. AB - OBJECTIVE: In rheumatoid arthritis (RA), synovial fibroblasts express proteases such as collagenases or cathepsins and inflammatory cytokines at elevated levels and so contribute to the inflammatory degradation process. Extracellular matrix degradation and cathepsin activity is dependent upon the presence of an acidic milieu. We examined whether activated synovial fibroblasts secrete acidic components. METHODS: Synovial fibroblasts were isolated and immortalized to study the mechanisms of metabolic activation. Naive and immortalized fibroblasts were activated with different cytokines. The responses were investigated by immunoblot to detect Egr-1 and by a cytosensor microphysiometer analysis to evaluate acid secretion. Basic gene expression patterns were investigated in naive and immortalized cells by RT-PCR analysis. RESULTS: We found RA synovial fibroblasts respond to different cytokines associated with the pathomechanisms of RA including interleukin 1, basic fibroblast growth factor, platelet derived growth factor, and tumor necrosis factor-alpha, with metabolic activation and enhanced secretion of acidic components. In addition, naive and SV40 TAg immortalized fibroblasts rapidly release acidic components after stimulation with phorbol ester or ionomycin as well. CONCLUSION: Activated synovial fibroblasts not only express inflammatory cytokines and matrix degrading proteases that are associated with the pathomechanisms of RA, but upon stimulation may release acidic components that lower pH and consequently enhance cathepsin activity and collagen solubilization. PMID- 11036824 TI - Lag time between onset of symptoms and access to rheumatology care and DMARD therapy in a cohort of patients with rheumatoid arthritis. AB - OBJECTIVE: To study demographic and clinical variables associated with a longer delay in disease modifying antirheumatic drug (DMARD) therapy initiation in a cohort of patients with rheumatoid arthritis (RA). METHODS: We studied 527 new RA patients (74.3% female, median age at symptom onset 55 yrs) in a hospital setting who fulfilled the ACR criteria for the diagnosis of RA. Demographic, clinical, laboratory, and treatment variables were collected longitudinally into a computerized research database. Risk factors for delay in use of DMARD therapy and first evaluation by a rheumatologist were analyzed using a Cox regression model. RESULTS: The median lag time between symptom onset and first rheumatologist encounter was 17 months and between onset of symptoms and first DMARD therapy 19 months. Variables associated with longer delay to DMARD therapy were the lag time between symptom onset and first rheumatologist visit (RR 0.73, 95% CI 0.71-0.76) and years of education. Variables associated with longer delay in first visit with rheumatologist were swollen/tender joint count, age at symptom onset, home support, labor force status, marital status, and years of education. CONCLUSION: Awareness of factors associated with a longer delay in access to rheumatology care and DMARD therapy may help break down barriers that prevent their early access, irrespective of patient age, socioeconomic status, initial symptoms, or need for treatment. PMID- 11036825 TI - Cyclosporin A inhibits CD69 expression induced on synovial fluid and peripheral blood lymphocytes by interleukin 15. AB - OBJECTIVE: To study the modulation of CD69 expression on peripheral blood (PB) and synovial fluid (SF) lymphocytes by interleukin 15 (IL-15) and several other cytokines and chemokines widely detected in the rheumatoid microenvironment. The effect of cyclosporin A (CSA) or methotrexate (MTX) in the cytokine mediated regulation of CD69 was analyzed. METHODS: CD69 expression on lymphocytes was assessed by flow cytometry after incubation with different cytokines, chemokines, phorbol myristate acetate, or calcium ionophore in the presence or absence of CSA, MTX, or both. The effect of IL-15 and SF supernatants in maintaining CD69 expression on SF lymphocytes was also assessed. IL-15 levels in SF supernatants were measured by ELISA. RESULTS: IL-15 induced the greatest upregulation of CD69 expression on PB lymphocytes in a time and dose dependent manner. IL-15 was able to maintain a high CD69 expression on SF lymphocytes. SF supernatants from rheumatoid arthritis (RA), which contain significant amounts of IL-15, also reversed the CD69 downregulation of SF lymphocytes in culture. CSA, but not MTX, inhibited the CD69 upregulation mediated by IL-15 both in PB and SF lymphocytes. CONCLUSION: IL-15 appears to be responsible, at least in part, for the high CD69 expression on lymphocytes from the rheumatoid microenvironment. Consistent with the virtual absence of lymphocyte derived cytokines in RA synovium, the prevention of IL-15 mediated CD69 upregulation on lymphocytes may explain the effect of CSA in the treatment of RA. PMID- 11036826 TI - Imbalance in production between vascular endothelial growth factor and endostatin in patients with rheumatoid arthritis. AB - OBJECTIVE: To clarify whether synovial cell proliferation indicates an imbalance in production between angiogenic growth factors and angiogenesis inhibitors in rheumatoid arthritis (RA), we investigated the production of basic fibroblast growth factor (b-FGF) and vascular endothelial growth factor (VEGF) as representative angiogenic growth factors and endostatin as a representative angiogenesis inhibitor. METHODS: The b-FGF, VEGF, and endostatin levels in 90 samples of peripheral blood (PB) and 15 samples of joint fluid obtained from patients with RA and 30 samples of PB and 10 samples of joint fluid from patients without RA, including 20 patients with inflammatory arthritis without purulent arthritis, and 10 patients with osteoarthritis were measured by ELISA. VEGF and endostatin levels in blood samples from 22 patients with RA were measured at 2 points: before and 4 or 5 months after the commencement of medication. RESULTS: The b-FGF and VEGF levels in the PB and joint fluid samples from patients with RA were markedly elevated compared to samples from patients without RA. In contrast, endostatin levels in PB and joint fluid samples from patients with RA were almost the same as in the samples from patients without RA. VEGF levels in blood samples obtained 4 or 5 months after the commencement of medication (combination of prednisolone 5 mg/day and disease modifying antirheumatic drugs: either bucillamine 100 mg/day or salazosulfapyridine 1,000 mg/day) were significantly decreased from 27.1 +/- 8.5 pg/ml in samples obtained before commencement of medication to 18.1 +/- 16.2 pg/ml. Endostatin levels in the corresponding samples were significantly increased, from 31.5 +/- 7.0 to 57.1 +/- 22.8 ng/ml [correction]. CONCLUSION: Our results reveal significant differences in b-FGF and VEGF levels in PB and joint fluid samples, but no difference in endostatin levels, between patients with RA and those without RA, suggesting that angiogenesis in RA occurs as a result of an imbalance in production between angiogenic growth factors and angiogenesis inhibitors. PMID- 11036827 TI - Efficacy of fish oil concentrate in the treatment of rheumatoid arthritis. AB - OBJECTIVE: To determine the efficacy of fish oil derived (n-3) fatty acid supplementation (3-6 capsules/day) in subjects with rheumatoid arthritis (RA) whose (n-6) fatty acid intake in the background diet was < 10 g/day, compared to olive/corn oil capsule supplement over a 15 week period. METHODS: A placebo controlled, double blind, randomized 15 week study to determine the effect of supplementation on clinical variables in 50 subjects with RA whose background diet was naturally low in (n-6) fatty acids. Fish oil containing 60% (n-3) fatty acids was supplemented at a rate of 40 mg/kg body weight. RESULTS: Analysis of 9 clinical variables indicated there was a significant difference (p < 0.02) between control and treatment groups. Five subjects in the treatment group and 3 in the control group met the American College of Rheumatology 20% improvement criteria. Dietary supplementation resulted in a significant increase in eicosapentaenoic acid in plasma and monocyte lipids in the supplemented group. CONCLUSION: The findings suggest that fish oil supplementation that delivers (n 3) fatty acids at a dose of 40 mg/kg body weight/day, with dietary (n-6) fatty acid intake < 10 g/day in the background diet, results in substantial cellular incorporation of (n-3) fatty acids and improvements in clinical status in patients with RA. PMID- 11036828 TI - Expression of the precursor of secretoneurin, secretogranin II, in the synovium of patients with rheumatoid arthritis and osteoarthritis. AB - OBJECTIVE: Secretoneurin (SN) is a neuropeptide that is chemotactic for mononuclear cells and it has been suggested to be involved in the mediation of pain; there is also evidence that SN is involved in the inflammation process. As secretogranin II (SGII) is the precursor of SN, we investigated expression of SGII mRNA and SN protein in the synovium of patients with rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS: Snap frozen synovial tissue specimens from 12 patients with RA and 11 patients with OA were examined. RNA was isolated and cDNA copied by reverse transcription. The expression of SGII was determined by polymerase chain reaction and in situ hybridization (ISH). SGII expressing cells were compared with CD68 positive cells stained by immunohistochemistry. SN protein was also detected by immunohistochemistry. RESULTS: A 524 bp SGII specific fragment could be amplified by PCR from the cDNA of all specimens. ISH showed scattered expression of SGII in both RA and OA synovial tissue; its expression pattern was characterized by positive staining for SGII in both the lining and the sublining layer. Immunohistochemical double labeling with anti CD68 antibodies revealed expression of SGII in CD68 negative, fibroblast-like cells, whereas CD68 positive macrophages did not. In RA and OA, the SGII staining by ISH was positive with a diffuse staining throughout the entire synovial tissue. SN protein expression was scattered in RA but more intense in OA synovium. CONCLUSION: The expression of SGII mRNA in RA and OA synovial fibroblasts clearly supports the hypothesis that SN is involved in the synovial tissue inflammation in both diseases. The significant lower SN expression in RA could be due to an inhibitory mechanism with respect to the SN levels in synovial fluid. SN might be involved in the modulation of afferent nerve transmission and therefore might play a role in the sensation of pain, especially in patients with OA. PMID- 11036829 TI - The influence of age, sex, and race on the upper reference limit of serum C reactive protein concentration. AB - OBJECTIVE: The recommended reference range for serum C-reactive protein (CRP) concentrations is usually not adjusted for age and sex. We sought to determine if age, sex, and race or ethnicity influence the distribution of CRP values, and if upper reference limits of CRP should be adjusted by demographic factors. METHODS: Interviews, physical examinations, and blood draws were performed on > 22,000 individuals age > or = 4 yrs representative of the noninstitutionalized population of the United States, as part of the Third National Health and Nutrition Evaluation Survey (NHANES III). Serum CRP concentrations were measured by nephelometric immunoassay. RESULTS: The 95th percentile value of CRP in the overall population was 0.95 mg/dl for males and 1.39 mg/dl for females, and varied with age and race. For ages 25-70 yrs, the age adjusted approximate upper reference limit (mg/dl) was CRP = age/50 for males, and CRP = age/50 + 0.6 for females. The upper limits for Mexican-Americans and non-Hispanic whites were similar, whereas for non-Hispanic black adults the approximate upper limit was CRP = age/30 for males and CRP = age/50 + 1.0 for females. Even after accounting for identified inflammatory conditions, demographic factors influenced the reference limits of CRP. The 95th percentile values were uniformly lower in children than in older adults. CONCLUSION: Demographic factors, including age, sex, and race, should be used to adjust the upper reference limit for CRP. Clinicians should be aware of these factors when using CRP values to assess inflammatory diseases. PMID- 11036830 TI - One year followup variables predict disability 5 years after presentation with inflammatory polyarthritis with greater accuracy than at baseline. AB - OBJECTIVE: To compare the accuracy of simple demographic and clinical variables recorded at baseline with those recorded after one year followup, in predicting self-reported functional disability recorded 5 years after initial assessment in patients with early inflammatory polyarthritis (IP). METHODS: We followed annually for 5 years 528 patients registered by the Norfolk Arthritis Register (a primary care based cohort of patients with early IP) using the Health Assessment Questionnaire (HAQ). Backward stepwise logistic regression was used to determine the clinical and demographic variables, collected at either baseline or first followup, that were associated with disability (HAQ > or = 1.00/> or =1.50) at 5 years. RESULTS: At the 5th anniversary assessment, the prevalence of moderate disability (HAQ > or = 1.00) was 47%. Twenty-nine percent reported more severe disability (HAQ > or = 1.50). Variables recorded at first anniversary assessment were better able to predict patients at risk of developing a poor outcome than baseline variables. Multivariate methods identified age at symptom onset, HAQ score, presence of nodules, and a statistically derived factor describing joint tenderness recorded at first year as important predictors of both moderate disability (HAQ > or =1.00) and a higher level of disability (HAQ > or = 1.50). When tested in an independent validation sample, the accuracy of the models generated from data recorded at the first year was 76% (HAQ > or = 1.00) and 83% (HAQ > or = 1.50). CONCLUSION: It was possible to predict disability at 5 years with high accuracy using simple clinical variables and demographic data collected 4 or 5 years previously. First year HAQ score was the strongest predictor of future disability. HAQ score at 5 years could be predicted more accurately using data collected at first anniversary visit than using data recorded at baseline. PMID- 11036831 TI - Evaluation of predictive factors for neurocognitive dysfunction in patients with inactive systemic lupus erythematosus. AB - OBJECTIVE: To determine predictive factors associated with the cognitive dysfunction in patients with inactive systemic lupus erythematosus (SLE). METHODS: Consecutive patients followed at the Lupus Clinic with inactive SLE (SLE Disease Activity Index, SLEDAI, = 0) underwent a battery of neuropsychological tests; Beck Depression Inventory and psychiatric assessment were also performed. Neurocognitive dysfunction was defined as 3 abnormal scores. Data were analyzed using chi-square tests, ANOVA tests, and logistic regression. RESULTS: Twenty five of the 58 patients with SLE (43%) versus 9 of 47 healthy controls (19%) demonstrated neurocognitive dysfunction (p < 0.01). Neurocognitive dysfunction was not associated with depression or a psychiatric diagnosis, use of steroids, or previous or current evidence for fibromyalgia. SLEDAI > 10 at first presentation to the Lupus Clinic and previous vasculitis were associated with neurocognitive dysfunction, but previous central nervous system disease, renal disease, renal damage, or atherosclerotic complications were not. Neurophysiologic studies at the time of the assessment were not predictive of neurocognitive dysfunction. CONCLUSION: Patients with inactive SLE demonstrate neurocognitive dysfunction. This is associated with more disease activity at presentation, but is not associated with specific organ involvement or organ damage. PMID- 11036832 TI - Single photon emission computed tomography dual isotope myocardial perfusion imaging in women with systemic lupus erythematosus. I. Prevalence and distribution of abnormalities. AB - OBJECTIVE: To determine the prevalence of myocardial perfusion abnormalities in women with systemic lupus erythematosus (SLE) using single photon emission computed tomography (SPECT) dual isotope myocardial perfusion imaging (DIMPI). METHODS: Consecutive female patients registered at the University of Toronto Lupus Clinic were offered DIMPI evaluation and all who accepted were studied. Patients underwent SPECT DIMPI using dipyridamole stress. Resting and stress images were acquired using thallium-201 (201TI) and technetium 99m-sestamibi (99mTc sestamibi), respectively. We recorded segmental perfusion abnormalities, severity and reversibility of any abnormality, and number of vessel territories involved. Ejection fraction was also measured. RESULTS: One hundred thirty patients were studied. Mean (SD) age and disease duration at study were 45.1 (11.1) years and 14.6 (9.4) years, respectively. Thirteen patients (10%) had a history of angina pectoris or myocardial infarction. Overall, 52 (40%) patients had an abnormality of myocardial perfusion, including 11 (85%) with a history of angina or myocardial infarction. In those with no history of coronary artery disease, 41 (35%) had an abnormality detected. The perfusion defect was reversible in 47 (90%). In 37 (71%) cases perfusion defects were seen in the region of a single vessel territory. Eighteen (13.8%) patients had an ejection fraction (EF) < 50%. CONCLUSION: Using SPECT DIMPI, 40% of all women with SLE and 35% of women with SLE with no history of coronary artery disease had abnormalities of myocardial perfusion, suggesting a high prevalence of early coronary artery disease. The early detection of disease will facilitate study of atherosclerotic risk factors; such women can also be targeted for a focused program of risk factor management. PMID- 11036833 TI - Large macrophage colony-forming cells identical to high proliferative potential colony-forming cells in peripheral blood of patients with collagen vascular diseases: high occurrence among patients with systemic sclerosis and dermatomyositis. AB - OBJECTIVE: To examine peripheral blood (PB) of patients with various collagen vascular diseases (CVD) for the presence of colony-forming cells (CFC) that form large macrophage colonies (> 2.5 mm in diameter, > 10,000 cells). METHODS: Peripheral blood mononuclear cells were obtained from 92 patients with various active CVD and 20 healthy controls, and assayed for in vitro colony formation. There were 14 patients with systemic lupus erythematosus (SLE), 30 with rheumatoid arthritis (RA), 17 with systemic sclerosis (SSc), 20 with polymyositis (PM)/dermatomyositis (DM) (11 PM, 9 DM) and 11 with systemic vasculitis. RESULTS: Large macrophage CFC were detected in PB of 7% of patients with SLE (1/14), 17% with RA (5/30), 47% with SSc (8/17), 30% with PM/DM (6/20) [9% PM (1/11) and 56% DM (5/9)], 0% of those with systemic vasculitis (0/11) and 0% of the healthy subjects (0/20). There was a significant difference between the occurrence of CFC in patients with PM versus patients with DM (p < 0.05). The occurrence of CFC in patients with SSc or DM was significantly higher than that in patients with other CVD including SLE, RA, PM, and systemic vasculitis (p < 0.05). CONCLUSION: Based on the size of the colonies they formed, the CFC corresponded to high proliferative potential colony-forming cells, a subset of primitive hematopoietic cells. Our findings among patients with CVD indicate that these primitive hematopoietic progenitor cells, which are believed to constitute a noncirculating population in healthy individuals, are found most frequently in PB of patients with SSc and DM. It is likely that primitive hematopoietic cells are frequently mobilized into the peripheral circulation during the pathogenesis of SSc and DM. PMID- 11036834 TI - Antibodies to AB blood group antigens mimic anti-salivary duct autoantibodies in patients with limited sicca symptoms. AB - OBJECTIVE: We evaluated the clinical relevance and pathogenic significance of anti-salivary duct autoantibodies (ASDA) in Sjogren's syndrome (SS) and rheumatoid arthritis (RA) by examining (1) their frequency in healthy controls, patients with sicca symptoms, and patients with various autoimmune and infective disorders; (2) their localization by confocal microscopy; and (3) their tissue distribution and cross reactivity with blood group antigens. METHODS: Indirect immunofluorescence (IF) was performed on commercial cryostat sections of monkey parotid salivary gland. Sections were examined by fluorescence and confocal laser scanning microscopy. Sera giving positive staining on the ducts were tested by IF on a range of monkey tissues and salivary glands from several mammalian species. Blocking experiments were performed with human erythrocytes of different ABO blood groups and AB antigens. RESULTS: We identified 2 distinct ductal staining patterns. The first resembled ASDA described in earlier studies and showed patchy bright staining of the apical (luminal) surfaces of the ducts and staining of apical cytoplasmic vesicles. The other was only observed with anti-mitochondrial antibody positive sera and stained the mitochondrial-rich ductal epithelium in a distinctive punctate pattern. Antibodies staining the apical surface of ducts were detected rarely in patients with antiRo/La autoantibody-positive primary SS (1/76) and RA (1/36) and were found in only 1115 with RA and secondary SS. ASDA were detected in sera from 13/51 (25.5%) of patients referred to our clinic with limited sicca symptoms who were anti-Ro/La antibody-negative and had no typical clinical or laboratory features of classical primary SS. The apical ductal staining pattern was not observed with sera from 63 healthy controls without sicca symptoms or in patients with autoimmune and infective disorders. Twelve of the 13 patients whose sera gave ASDA-like staining were blood group O and one group A. Ductal staining was abolished in all sera after absorption with blood group AB erythrocytes or AB antigen. In 5 patients ductal staining was removed by absorption with B erythrocytes but not with A erythrocytes; in the remainder ductal reactivity was abolished by both A and B erythrocytes. CONCLUSION: ASDA seem to occur rarely in patients with primary SS and RA. However, isotype switched IgG AB blood group antibodies cross react with primate salivary ducts and may produce false positive ASDA staining. Detection of ASDA may be of value in identifying a subset of patients who present with mild sicca symptoms without other autoimmune features. PMID- 11036835 TI - Treatment of established collagen induced arthritis with prostaglandin E1 incorporated in lipid microspheres. AB - OBJECTIVE: In view of evidence obtained from in vitro and in vivo experiments that prostaglandin E1 (PGE1) has regulatory effects on disordered immune responses and inflammation, we investigated whether lipo-PGE1, an efficient drug delivery system incorporating PGE1 into lipid microspheres, can ameliorate arthritis in the collagen induced arthritis (CIA) model of rheumatoid arthritis (RA). METHODS: DBA/1J male mice were immunized with bovine type II collagen in adjuvant, and treated daily from onset of clinical arthritis with intravenous administration of lipo-PGE1 (5-50 microg/kg) or lipid vehicle as a control. Arthritis was assessed over a 10 day treatment period by monitoring for paw swelling and clinical score. Histopathology of the arthritic hind paws was also evaluated. Lipo-PGE1 accumulation in arthritic joint tissues was measured using 3H labeled PGE1 incorporated in lipid microspheres. RESULTS: Arthritis was significantly suppressed in lipo-PGE1 treated mice compared with lipid vehicle treated controls (p < 0.05, p < 0.016, respectively) in a dose-dependent manner. Histopathological assessment showed a significant reduction of pannus formation and joint destruction in lipo-PGE1 treated mice compared with controls (p < 0.05). Lipo-PGE1 preferentially accumulated in arthritic joints for a longer period than free PGE1. CONCLUSION: Using an efficient drug delivery system, PGE1 can suppress CIA, and lipo-PGE1 may have a potential therapeutic role in RA. PMID- 11036836 TI - Fas and Fas ligand gene polymorphisms in primary Sjogren's syndrome. AB - OBJECTIVE: To screen for polymorphisms in the apoptosis regulating Fas and Fas ligand (FasL) genes in patients with primary Sjogren's syndrome (SS), and to explore associations with susceptibility to the disease. METHODS: Polymorphisms in Fas and FasL of 70 patients with primary SS and 72 controls were determined by polymerase chain reaction combined with the restriction enzyme fingerprinting single strand conformation polymorphism technique, verified by automatic sequencing and natural or amplification created restriction site tests. RESULTS: Polymorphisms were found in both Fas and FasL, but only some of the Fas polymorphisms were found in statistically significant differences between patients and controls. Patients displayed a higher frequency of the G/G genotype at position -671 than the controls, and the -671 G allele frequency for primary SS was increased compared to controls. A higher frequency of the C allele at position IVS2nt176 and IVS5nt82 was also found. Of note, the nucleotide variants in intron 2 and intron 5 were associated. CONCLUSION: We describe the positions and frequencies of several polymorphisms in the genes encoding Fas and FasL in patients with primary SS. None caused any amino acid change. Three Fas alleles, of which one is located in the promoter area, showed significant although modest differences between patients and controls. PMID- 11036837 TI - De novo synthesis of proteinase 3 by cytokine primed circulating human polymorphonuclear neutrophils and mononuclear cells. AB - OBJECTIVE: When polymorphonuclear neutrophils (PMN) and peripheral blood monocytes (PBMC) are stimulated with tumor necrosis factor alpha (TNF-alpha), preexisting granule stored proteinase 3 (PR3) is translocated to the surface of their plasma membrane. We investigated whether PR3 gene reactivation and new PR3 protein production were also features of priming by cytokine. METHODS: Normal human PMN and PBMC were isolated and stimulated in vitro with TNF-alpha. They were harvested at different intervals and subjected to total RNA and protein analysis. PR3 mRNA was identified by reverse transcription polymerase chain reaction, Northern blot, and sequencing. De novo PR3 synthesis was evaluated by metabolic labeling with [35S] methionine followed by immunoprecipitation using anti-neutrophil cytoplasmic antibodies from serum of patients with active Wegener's granulomatosis and mouse monoclonal anti-native PR3 antibodies. RESULTS: Resting PMN and PBMC do not express PR3 mRNA. During priming, PR3 mRNA appears in PMN at 2 h, peaks at 6 h, and has disappeared at 12 h. By comparison, in primed PBMC, PR3 mRNA appears at 6 h, peaks at 12 h, and disappears at 24 h. Immunoprecipitation of metabolically labeled PR3 revealed new synthesis of PR3 by both cell types, a process that was inhibited by cycloheximide. CONCLUSION: Primed PMN and PBMC can express PR3 mRNA and synthesize new PR3 protein, providing an alternative source to membrane PR3. Whether that small amount of inducible PR3 has a primary structure, a localization, or a role different from those of preformed PR3 stored in granules remains to be clarified. PMID- 11036839 TI - Rheumatologic manifestations of pachydermoperiostosis and preliminary experience with bisphosphonates. AB - OBJECTIVE: Pachydermoperiostosis is manifested by finger clubbing, hypertrophic skin changes, and periosteal bone formation. We describe 5 cases revealed primarily by their rheumatologic manifestations. Also reported are preliminary experiences on the use of intravenous pamidronate as a treatment. METHODS: This is a retrospective study including the analysis of clinical manifestations, laboratory results and morphological examinations gathered from patients' medical records. We evaluated efficacy of treatment with 1 mg/kg iv pamidronate in the 3 new cases. RESULTS: Before treatment with iv pamidronate, the patients' global assessment was poor (twice) and very poor (once). The physician's global assessment was poor in 3 patients. After treatment with iv pamidronate, 2 patients had significant improvement. Physician and patient global assessments were very good, good, and moderate. No side effects were observed. All biological variables were within normal ranges at 12 month followup visit. CONCLUSION: Pachydermoperiostosis must be recognized by the rheumatologist, since it can present symptomatically through articular manifestations. When conventional treatment modalities fail, iv pamidronate might be useful. PMID- 11036838 TI - Change in diagnosis among orthopedists compared to non-orthopedists in the management of acute knee injuries. AB - OBJECTIVE: Uncertainty regarding diagnosis is associated with lower patient satisfaction and can lead to delays in definitive treatment and to inappropriate use of resources. We sought to compare change in diagnosis among orthopedists and non-orthopedists caring for a community based cohort of individuals with incident acute knee injuries. METHODS: We conducted a longitudinal investigation of a population based cohort of Olmsted County residents with their first episode of acute knee injury occurring between January 1, 1993, and December 31, 1995. We reviewed the entire (inpatient and outpatient) medical records for these patients and collected extensive clinical data on all diagnoses made (including possible and probable) and the specialty of the attending physician(s) making them. Diagnoses were categorized as: (1) meniscus injury, cruciate injury, or osteochondral fracture; (2) ligament injury, patellar instability, patellar injury; or (3) sprain, strain, injury (unspecified). Diagnostic switches were defined as changes from one diagnostic category to another, or the addition or subtraction of a diagnostic category. We then examined the quality of the documented evidence supporting meniscal, ligamentous, and cruciate diagnoses (at initial evaluation) by comparing the clinical evidence to the recommendations outlined by the American Academy of Orthopaedic Surgeons clinical algorithm on acute knee injury. Analyses were conducted comparing (1) the number of diagnostic switches and (2) the quality of the documented evidence among those cases initially cared for by orthopedists and those cared for by non-orthopedists, using logistic regression analysis adjusting for age, sex, and injury severity. The influence of these variables on costs of care was also examined. RESULTS: There were 664 patients (361 men and 303 women) in our study population, with an average age of 36.0 years (minimum 17, maximum 87). Of these, 324 were excluded because they only had one clinical encounter for their acute knee injury. Of the remaining 340, 59 (17.4%) were initially cared for by an orthopedist and 211 (62.1%) were cared for by an orthopedist at some time during their care. Diagnostic switches were significantly less frequent in the group who were cared for by orthopedists (55% vs 74%, p < 0.001). This result persisted after adjusting for age, sex, and severity (p = 0.003). The proportion of cases whose diagnoses were supported by evidence was significantly higher among the group whose first attending physician was an orthopedist (63.0% vs 37.6%, p = 0.002). Both change in diagnosis (p < 0.001) and physician specialty (p < 0.001) were statistically significant predictors of costs of care. CONCLUSION: Compared to non-orthopedic care, orthopedic care for acute knee injury was associated with fewer changes in diagnosis, and diagnoses made by orthopedists were more likely to be supported by evidence. However, even after adjusting for severity, orthopedic care remained significantly more costly than non-orthopedic care. PMID- 11036841 TI - Localized monocyte chemotactic protein-1 production correlates with T cell infiltration of synovium in patients with psoriatic arthritis. AB - OBJECTIVE: To examine normal and psoriatic skin and synovial tissue from patients with psoriatic arthritis (PsA) for evidence of monocyte chemotactic protein-1 (MCP-1) mediated T cell chemotaxis. METHODS: Peripheral blood (PB), synovial fluid (SF), normal and psoriatic skin, and synovial biopsies were obtained from patients with PsA (n = 19) and compared to samples from normal (n = 5) and disease (n = 5) controls (NC, DC). Immune cell populations in PB and SF samples were assessed by immunofluorescent labeling and flow cytometry, levels of soluble MCP-1 were determined by quantitative ELISA, and immunohistochemistry was used to detect T cell subsets and macrophages and MCP-1 protein in frozen skin and synovial tissue sections. RESULTS: CD8+ but not CD4+ T cells were elevated in SF compared to PB, and the majority of these cells expressed CD45RO. Plasma MCP-1 levels in PsA were elevated relative to NC. MCP-1 levels were significantly higher than paired plasma samples in patients with recent onset (< 6 mo) synovitis (n = 10). A positive correlation was observed between synovial T cell numbers and MCP-1 levels in SF. MCP-1 protein was present in all tissues examined, but most intense expression was observed in synovium. CONCLUSION: Elevated concentrations of MCP-1 concomitant with memory T cell infiltration in PsA SF suggests that MCP-1 mediated chemotaxis is involved in the recruitment of T lymphocytes into the synovial compartment of patients with PsA. PMID- 11036840 TI - A pooled data analysis on the use of intermittent cyclical etidronate therapy for the prevention and treatment of corticosteroid induced bone loss. AB - OBJECTIVE: To conduct a pooled data analysis in a group of patients defined by sex, menopausal status, and underlying disease in order to examine the effect of intermittent cyclical etidronate in the prevention and treatment of corticosteroid induced osteoporosis. METHODS: We selected 5 randomized, placebo controlled studies that examined the efficacy of intermittent cyclical etidronate therapy in which the raw data were available for analysis. Three were prevention studies and 2 treatment studies. The primary outcome was the difference between treatment groups in the percentage change from baseline in lumbar spine bone density. Secondary outcomes included the difference between treatment groups in the percentage change from baseline in femoral neck and trochanter bone density, and vertebral fracture rates. RESULTS: Results are separately pooled for the prevention and treatment studies. The prevention studies had significant mean differences (95% CI) between groups in mean percentage change from baseline in lumbar spine, femoral neck, and trochanter bone density of 3.7 (2.6 to 4.7), 1.7 (0.4 to 2.9), and 2.8% (1.3 to 4.2) after one year of treatment, in favor of the etidronate group. The treatment studies displayed a mean difference between groups in mean percentage change from baseline in lumbar spine bone density of 4.8 (2.7 to 6.9) and 5.4% (2.5 to 8.4) after one and 2 years of therapy. In the prevention studies, a reduced fracture incidence was observed in the etidronate group compared with the placebo group (relative risk 0.50; CI 0.21 to 1.19). CONCLUSION: Etidronate therapy was effective in preventing bone loss in the prevention studies and in preventing or slightly increasing bone mass in the treatment studies. A fracture benefit was observed in postmenopausal women treated with etidronate in the prevention studies. PMID- 11036842 TI - Parvovirus arthropathy outbreak in southwestern United States. AB - OBJECTIVE: We describe an outbreak of parvovirus (PV) arthropathy that was detected in a rheumatology clinic in San Antonio, Texas, during the winter of 1994. Parvovirus B19 causes acute symmetric polyarthritis (ASPA) in adults. In the US, the majority of cases described are from the northern US. METHODS: An outbreak of PV arthropathy was monitored in a San Antonio area rheumatology clinic. RESULTS: Of the 16 affected patients, 69% were female, ages ranging from 23 to 60 years; 75% had close contact with children, 58% of whom were exposed to children with clinical PV. All patients noted an acute arthritis except for 2 patients with polyarthralgias. The most common presentation was ASPA (9/16), with 10/16 complaining of viral prodrome, and 5/16 having a nonspecific rash, but none with the typical "slapped cheek" appearance. Eleven patients had an ASPA at some time in their illness. Of these, 3 had a true migratory arthritis that developed into an ASPA and another 2 were additive. Two additional patients had persistent asymmetric polyarthritis. The most common joints involved were the metacarpophalangeals, proximal interphalangeals, wrists, and knees. Most patients' syndromes lasted < 6 weeks, but 3 patients had symptoms that lasted longer than 6 months. Eight of 10 had elevated erythrocyte sedimentation rate. Rheumatoid factor was detected in 3 patients and antinuclear antibody in 2. All patients were treated symptomatically with nonsteroidal antiinflammatory drugs and a few also received low dose corticosteroids. Because of suspicious clinical presentations, 2 patients were presumed to have gonococcal arthritis before PV titers were available. CONCLUSION: This is the first large series on adults with PV arthropathy reported in the southern US. In contrast to the usual features of ASPA, the outbreak appears unique in that almost 40% of cases presented with a true migratory arthritis. PMID- 11036843 TI - A decline in lower extremity lean body mass per body weight is characteristic of women with early phase osteoarthritis of the knee. AB - OBJECTIVE: Sarcopenia progresses with aging, but the effect of muscle loss on degenerative joint disorders has not been precisely evaluated. We assessed the distribution of lean body mass (LBM) in middle aged women who had osteoarthritis (OA) of the knee for < 5 years. METHODS: Two hundred thirty-five Japanese women aged 45-69 years, including 117 with knee OA for < 5 years, and 118 age and sex matched healthy controls were studied. Body composition measurements were carried out by segmental bioelectrical impedance using 8 surface electrodes. Anthropometric data consisting of LBM of upper extremities/body weight (U-LBM/W), LBM of trunk/body weight (T-LBM/W), and LBM of lower extremities/body weight (L LBM/W) for knee OA participants were compared to corresponding data for controls. In the knee OA group, the significance of correlations between anthropometric data and Lequesne index of severity for knee OA and between anthropometric data and disease duration were assessed. RESULTS: L-LBM/W was significantly lower in knee OA participants compared with controls (19.2 +/- 2.7% vs 21.0 +/- 2.9%; p < 0.0001). However, no significant differences in U-LBM/W and T-LBM/W were observed between the knee OA and control groups (p > 0.2). L-LBM/W did not correlate significantly with the index of severity or disease duration (p > 0.2). CONCLUSION: Decline in L-LBM/W, but not U-LBM/W or T-LBM/W, may be important in the pathogenesis of knee OA. PMID- 11036844 TI - Extracorporeal shock wave application for chronic plantar fasciitis associated with heel spurs: prediction of outcome by magnetic resonance imaging. AB - OBJECTIVE: To clarify morphologic features associated with the clinical outcome of extracorporeal shock wave application (ESWA) in chronic plantar fasciitis. METHODS: In this prospective study 43 patients (48 heels) with chronic courses of plantar fasciitis were clinically examined before and after repetitive low energy ESWA. Standard radiographs of the affected heels were obtained before ESWA to document the existence of a calcaneal heel spur. Magnetic resonance imaging (MRI) was performed before ESWA to evaluate abnormalities of the plantar fascia, the surrounding soft tissue structures, and bone marrow edema of the calcaneus. RESULTS: After ESWA (mean followup 19.3 mo), clinical evaluation of all 48 heels revealed a statistically significant decrease in the mean visual analog scale score from 74.5 to 25.4. Using the Roles and Maudsley score (RM), an established scoring system for categorizing results of treatment following ESWA for patients with plantar fasciitis, patients could be divided into 2 groups, i.e., satisfactory clinical outcome of ESWA (grades 1 and 2 by RM scale; n = 36 heels) and unsatisfactory outcome (grades 3 and 4 by RM scale; n = 12 heels). While thickness of plantar aponeurosis, soft tissue signal intensity changes, and soft tissue contrast medium uptake did not correlate with clinical outcome, the presence of a calcaneal bone marrow edema was highly predictive for satisfactory clinical outcome (positive predictive value 0.94, sensitivity 0.89, specificity 0.8). CONCLUSION: This study indicates that in patients with chronic plantar fasciitis, the presence of calcaneal bone marrow edema on pretherapeutic MRI is a good predictive variable for a satisfactory clinical outcome of ESWA. PMID- 11036845 TI - The inhibition of spontaneous and tumor necrosis factor-alpha induced neutrophil apoptosis by crystals of calcium pyrophosphate dihydrate and monosodium urate monohydrate. AB - OBJECTIVE: Spontaneous neutrophil apoptosis may be inhibited by various proinflammatory stimuli. which may result in prolonged lifetimes and responses of these phagocytic cells with the potential for extended inflammation. We investigated the effect of short term incubation of opsonized crystals of monosodium urate monohydrate (MSUM) or calcium pyrophosphate dihydrate (CPPD) on both spontaneous and tumor necrosis factor-alpha (TNF-alpha) induced neutrophil apoptosis. METHODS: Peripheral neutrophils were incubated with plasma opsonized crystals of CPPD or MSUM in the presence or absence of TNF-alpha for 4 h at 37 degrees C. Apoptosis was determined using 3 separate assays: (1) an agarose DNA fragmentation assay, (2) a cytoplasmic histone associated DNA fragmentation assay, and (3) a caspase 3 fluorometric assay. RESULTS: All 3 assays showed similar results. Both MSUM and CPPD crystals inhibited spontaneous apoptosis in neutrophils. TNF-alpha induced high levels of apoptosis in neutrophils. However, co-incubation of the cells with TNF-alpha and crystals resulted in the inhibition of apoptosis to levels below those of control cells. Pretreatment of neutrophils with the protein synthesis inhibitor cycloheximide prevented the inhibition of apoptosis by crystals. CONCLUSION: These data support the concept of crystal induced inhibition of neutrophil apoptosis as part of the pathophysiology of the diseases collectively known as crystal induced arthritis. PMID- 11036846 TI - Pool exercise combined with an education program for patients with fibromyalgia syndrome. A prospective, randomized study. AB - OBJECTIVE: To evaluate the effects of 6 months of pool exercise combined with a 6 session education program for patients with fibromyalgia syndrome (FM). METHODS: The study population comprised 58 patients, randomized to a treatment or a control group. Patients were instructed to match the pool exercises to their threshold of pain and fatigue. The education focused on strategies for coping with symptoms and encouragement of physical activity. The primary outcome measurements were the total score of the Fibromyalgia Impact Questionnaire (FIQ) and the 6 min walk test, recorded at study start and after 6 mo. Several other tests and instruments assessing functional limitations, severity of symptoms, disabilities, and quality of life were also applied. RESULTS: Significant differences between the treatment group and the control group were found for the FIQ total score (p = 0.017) and the 6 min walk test (p < 0.0001). Significant differences were also found for physical function, grip strength, pain severity, social functioning, psychological distress, and quality of life. CONCLUSION: The results suggest that a 6 month program of exercises in a temperate pool combined with education will improve the consequences of FM. PMID- 11036847 TI - Stringent endocrinological testing reveals subnormal growth hormone secretion in some patients with fibromyalgia syndrome but rarely severe growth hormone deficiency. AB - OBJECTIVE: Several reports suggest that growth hormone (GH) deficiency may be a pathogenic factor in fibromyalgia syndrome (FM). This hypothesis has never been adequately examined. METHODS: We measured serum GH concentration after insulin induced hypoglycemia in subjects with FM. GH secretion in subjects with a maximal GH increase < 10 ng/ml after hypoglycemia was assessed by additional arginine stimulation. RESULTS: In one of 56 subjects tested, GH remained below 3 ng/ml in both tests, satisfying the criteria for adult GH deficiency. Thirty-two subjects (67%) had a maximal GH > 10 ng/ml. We retrospectively found an inverse correlation between low density lipoprotein levels and maximal GH concentration in a subgroup of patients. CONCLUSION: Severe GH deficiency is not a significant pathogenic factor in most patients with FM. We observed an impaired reactivity of the somatotropic axis in one-third of patients with FM, in keeping with a functional alteration of the hypothalamus. PMID- 11036848 TI - Relationship between sex and antibodies to high mobility group proteins 1 and 2 in juvenile idiopathic arthritis. AB - OBJECTIVE: To compare the frequencies of antibodies to high mobility group proteins 1 and 2 (HMG-1, HMG-2) in boys and girls with juvenile idiopathic arthritis (JIA). METHODS: Sera of 60 children (44 girls, 16 boys) with JIA were screened for the presence of anti-nuclear antibodies (ANA) and antichromatin antibodies by indirect immunofluorescence (IIF) on eukaryotic cells and were assayed further for the presence of antibodies to purified HMG-1 and HMG-2 by enzyme immunoassays. RESULTS: A positive test for ANA was significantly associated with the presence of antibodies to both HMG-1 and HMG-2. There was a significant association between antibodies targeting the chromosomal regions of metaphase cells and antibodies to both HMG-1 and HMG-2. Females were significantly more likely than males to have ANA, and specifically more likely to have antibodies to HMG-1. There was a significant association between the presence of antibodies to HMG proteins and chromosomal reactivity detected by IIF on HEp-2 cells. CONCLUSION: The results suggest that females with JIA are more likely to be ANA positive than males and more likely than males to have antibodies to HMG-1. PMID- 11036849 TI - Intravenous immunoglobulin therapy for children with Stevens-Johnson syndrome. AB - OBJECTIVE: To review the experience with intravenous immunoglobulin (IVIG) therapy in patients with Stevens-Johnson syndrome (SJS) in our institution. METHODS: All charts of patients with SJS admitted to Children's Hospital between November 1988 and June 1998 were reviewed. RESULTS: Twelve patients with SJS were detected. There were 8 males and 4 females, with a mean age 6 years (range 10 mo to 17 yrs). All patients presented with high fever and cutaneous and mucous membrane changes, and the diagnosis SJS was confirmed by a dermatologist. Of the 12 patients with SJS, 7 were treated with IVIG, 2 with corticosteroids, and 3 with supportive care. IVIG was administered in a single infusion at 1.5-2 g/kg, and was given on an average of hospital day 3 (range 1-8 days). The average duration of fever was 8 days (range 3-14) in the IVIG treated patients compared to 14 days (range 6-20) in the non-IVIG treated group (p = 0.06). The mean hospital stay was 12 days (range 4-22) for the patients treated with IVIG and 15 days (range 6-25) for the non-IVIG treated group (p = 0.5). No toxicity was observed with IVIG therapy. CONCLUSION: Duration of fever was shortened in patients treated with IVIG, although statistical significance was marginal. The hospital stay was slightly shortened in patients treated with IVIG; however, statistical significance was not reached. Prospective and controlled, multicenter studies are needed to further investigate these preliminary findings. PMID- 11036850 TI - Intravenous immunoglobulin therapy for juvenile dermatomyositis: efficacy and safety. AB - OBJECTIVE: To assess the efficacy of intravenous immunoglobulin (IVIG) for the treatment of juvenile dermatomyositis (JDM) in patients who were unresponsive to corticosteroids (steroid resistant or steroid dependent) or showed unacceptable toxicity. METHODS: A retrospective chart review of the course of all patients with JDM treated with IVIG who attended the Dermatomyositis Clinic at The Hospital for Sick Children, Toronto, Canada, from August 1986 to December 1996. RESULTS: Eighteen patients with JDM were treated with IVIG. Ten patients were taking additional 2nd line treatments, methotrexate, azathioprine, cyclosporine, and cyclophosphamide. The main indication for starting IVIG was the failure of steroid therapy to induce remission of JDM. Twelve patients showed clinical improvement with IVIG. In these patients, the corticosteroid dose was reduced by > 50% for > 3 months without clinical or biochemical flare. Nine of these 12 patients had IVIG alone as a 2nd line agent, whereas 3 patients were treated with additional agents. Six patients remained steroid dependent; they subsequently required multiple agents to induce remission of JDM. CONCLUSION: Most steroid dependent or steroid resistant patients in our clinic were able to markedly reduce their dose of corticosteroid with the addition of IVIG. Given the retrospective nature of our data and the fact that multiple agents were sometimes used together, it will be important to confirm these findings in a controlled trial. PMID- 11036851 TI - Polyarteritis nodosa mimicking prostatic cancer. AB - We describe a 72-year-old man with prostate enlargement, prostate-specific antigen level of 35 ng/dl, mild polyarthritis, and constitutional symptoms. Prostatic ultrasonography suggested neoplasm; however, transrectal biopsy revealed findings consistent with polyarteritis nodosa (PAN). The patient went on to develop leg paresthesia and dysesthesia, increased serum creatinine, and systemic hypertension. Steroids and intravenous cyclophosphamide were administered, followed by improvement. Our case emphasizes the protean onset of PAN, and provides a new differential diagnosis of prostatic diseases related to elevated prostate-specific antigen. PMID- 11036852 TI - Case series: coexistence of Sjogren's syndrome and sarcoidosis. AB - OBJECTIVE: To present cases of coexisting sarcoidosis and Sjogren's syndrome (SS). METHODS: We analyzed the clinical data of our 464 patients with SS with a clinical picture suggesting coexisting SS and sarcoidosis. We followed them for several years. RESULTS: We found 5 patients with coexisting SS and sarcoidosis. In 3 patients, pure sarcoidosis could mimic SS. CONCLUSION: In our experience, the frequency of sarcoidosis in SS is much higher than in the general population. One of the European criteria is that before the diagnosis of SS is made, the presence of sarcoidosis must be excluded. From our experience of 5 cases we determined that the 2 diseases can coexist, which indicates the value of these criteria. PMID- 11036853 TI - Wegener's granulomatosis: pitfalls in the management of pulmonary disease: A case of Wegener's granulomatosis with a hilar mass. AB - Wegener's granulomatosis (WG) is a systemic, granulomatous vasculitis that typically affects the upper airways, lungs, and, in most cases, the kidneys. Lung involvement occurs in 85% of patients. A classic feature of WG is multiple pulmonary nodules, which frequently cavitate. Hilar adenopathy or mediastinal masses are rare. These atypical pulmonary findings should raise suspicion of diseases other than WG and lead to biopsy with cultures, even when the diagnosis of WG appears to be certain. These guidelines proved to be reliable in a patient with WG in whom a hilar mass was associated with tuberculosis. PMID- 11036854 TI - Spinal hyperostosis--a rare skeletal manifestation of psoriasis vulgaris. AB - A 28-year-old woman with uncomplicated psoriasis vulgaris presented with spinal hyperostosis and osteitis. The absence of peripheral arthritis, sacroiliitis, and diffuse idiopathic skeletal hyperostosis (DISH) raised the possibility that the spinal lesion was an extracutaneous manifestation of psoriasis. We review the association between uncomplicated psoriasis vulgaris and skeletal involvement. PMID- 11036855 TI - Hypothenar hammer syndrome followed by systemic sclerosis. AB - We describe the first case of bilateral hypothenar hammer syndrome (HHS) followed by systemic sclerosis (SSc) that was associated with silica exposure (Erasmus syndrome). The patient was a woman smoothing tiles in an earthenware factory who presented with bilateral digital ischemia associated with Raynaud's phenomenon. HHS was diagnosed, based on an angiographic study showing aneurysm of the ulnar arteries and occlusions of multiple digital arteries. Pulmonary silicosis was also diagnosed on pulmonary tomodensitometry. Two years later digital swelling with acroosteolysis developed. The FANA test was positive (titer 1:640) and anticentromere antibody tests were also positive. Esophageal manometry showed dysmotility of the lower esophagus. These findings were consistent with a diagnosis of SSc. PMID- 11036856 TI - Cerebral aspergillosis in systemic lupus erythematosus. PMID- 11036857 TI - Pseudotumor of the knee in a kidney-pancreas transplant patient. PMID- 11036858 TI - Familial hypertrophic synovitis. PMID- 11036859 TI - Undergraduate education in the AFLAR region. PMID- 11036860 TI - Percutaneous vertebroplasty in the treatment of osteoporotic vertebral compression fractures: an open prospective study. PMID- 11036861 TI - Cold injury in 2 patients with connective tissue disease--frostbite arthritis plus. PMID- 11036862 TI - Effect of estrogen and progesterone on gene expression of growth regulatory molecules and proto-oncogene in vascular smooth muscle cells. AB - We examined influences of estrogen and progestogen on gene expression of the growth regulatory molecules: platelet-derived growth factor (PDGF), interleukin-1 (IL-1), interleukin-6 (IL-6) and proto-oncogene c-myc in vascular smooth muscle cells (VSMC) by reverse transcription-polymerase chain reaction (RT-PCR) and Southern-blotting. VSMC were exposed to estrone-sulfate (E1-S) and medroxyprogesterone acetate (MPA) to induce differentiation. E1-S inhibited the expression of PDGF-A chain, IL-1, IL-6 and c-myc mRNA, whereas MPA had no effect. Inhibition by E1-S was not affected by treatment combined with MPA. These findings suggest that estrogen modulates these growth regulatory molecules and c myc gene expression in VSMC but not progestogen. We concluded that estrogen may have a direct atheroprotective effect through inhibition of growth regulatory factors. PMID- 11036863 TI - Increase in serum ceruloplasmin with aging is not observed in type 2 diabetes. AB - Changes of serum ceruloplasmin (Cp) levels have been reported in many conditions including diabetes mellitus (DM), in which the serum Cp levels were increased. In this study, we have examined the influence of aging on serum Cp levels in normal individuals and in individuals with DM. Serum Cp levels were measured in 85 outpatients with type 2 diabetes (type 2 DM group) as well as in 71 healthy individuals (control group). All patients recruited for this study were negative for the glutamic acid decarboxylase (GAD) antibody. The subjects were sub-grouped based on their ages (<55 and 55 < or =). The serum Cp levels in the control group increased significantly with aging (r=0.325, p<0.0055), while levels in the type 2 DM group did not (r=0.091, p=0.4079). The levels in the type 2 DM group (<55) were significantly higher than those in the control group (<55) (p = 0.0029), while the Cp levels in the type 2 DM group (55 < or =) were not different from those in the control group (55 < or =) (p=0.4187). An age-related increase of serum Cp levels was observed in normal individuals, but this change was not observed in type 2 DM patients since serum Cp levels in type 2 DM patients of all ages were similar to the levels in normal elderly individuals. PMID- 11036864 TI - A defect of lipopolysaccharide-induced nitric oxide synthase gene expression in the paraventricular nucleus of Lewis rats. AB - The hypothalamo-pituitary-adrenal (HPA) axis activity of Lewis rats has been reported to be hyporesponsive to immune challenge, and nitric oxide (NO) has been demonstrated to be involved in the regulation of the HPA axis during the response to immune challenge. The present study investigates the effect of systemic injection of lipopolysaccharide (LPS) on NO production within the paraventricular nucleus (PVN) of immature female Fisher-344 (F344) and Lew/N rats (Lew/N), to clarify the pathophysiological role of NO in the dysregulation of the HPA axis in Lewis/N. Intraperitoneal injection of 25 mg/kg LPS significantly increased neuronal NO synthase (nNOS) mRNA expression in the PVN of F344 rats, but did not change nNOS mRNA levels in the PVN of Lew/N rats. CRF mRNA levels in the PVN significantly increased in response to LPS injection only in F344 rats. In contrast, inducible NOS (iNOS) mRNA increased similarly in both strains. These results demonstrated a defect of up-regulation of nNOS gene expression in the PVN of Lew/N rats following immune challenge. The defect appears to be associated with the dysfunction of the HPA axis in this strain. An increase in iNOS mRNA may partially restore NO production in the PVN. PMID- 11036865 TI - Evaluation of changes in bone density and biochemical parameters after parathyroidectomy in primary hyperparathyroidism. AB - We examined the relationships between serum levels of intact parathyroid hormone (PTH) and alkaline phosphatase (ALP) versus bone mineral density (BMD) at the lumbar spine and radius in terms of their preoperative values and of their annual percentage and net changes after parathyroidectomy (PTX) in 44 Japanese patients (14 men and 30 women) with primary hyperparathyroidism (pHPT). Lumbar and radial BMD values were measured by dual energy X-ray absorptiometry and single photon absorptiometry and were used for evaluating the cancellous and cortical bone mass, respectively. Age- and sex-adjusted value (Z-score) of the radial BMD was significantly lower than that of the lumbar BMD before and after PTX (P < 0.05). In preoperative patients, serum levels of both intact PTH and ALP were significantly and negatively correlated with Z-score of the radial BMD (P < 0.05 and P < 0.001, respectively), but not with that of the lumbar BMD. After PTX, serum levels of calcium, phosphorus, ALP, and PTH became normal, and both lumbar and radial BMD values markedly increased over 1 year, with percentage changes of 12.2+/-1.4% and 11.6+/-1.6%, respectively, which were larger than those in any other Caucasian study previously documented. Even in patients without osteopenia (Z-score of BMD 20), lumbar and radial BMD values increased considerably after the operation (9.6+/-1.9% and 6.7+/-1.4%, respectively). Annual percentage and net changes in lumbar BMD were significantly and negatively correlated with those in ALP with high correlation coefficients, but those in radial BMD were correlated only with the annual net change in ALP but not with the percentage change. No significant correlations were observed between annual changes in either lumbar or radial BMD and those in intact PTH. Taken together, this study shows that PTX causes dramatic improvements in both the cancellous and cortical bone mass in Japanese pHPT patients regardless of the severity of their osteopenia, and suggests that the cancellous and cortical bones react differently to a preoperative endogenous PTH excess and a high bone turnover rate as well as to the postoperative normalization of a bone turnover rate in the patients. PMID- 11036866 TI - Involvement of distinct signaling pathways in activin-induced increases in FSH secretion and enlargement of FSH cell population in the rat pituitary. AB - Activin-A induces increases in FSH secretion, as well as the number of immunoreactive FSH cells, in cultured rat pituitary cells. In this study, we examined whether mechanisms involved in these two actions of activin-A are identical or not, with respect to the involvement of cellular proliferation and of Ca2+-dependent signaling pathways. Treatment with activin-A (25 ng/ml) for 48 h caused increases in the number of cultured rat anterior pituitary cells that incorporated BrdU, a thymidine analog. The stimulatory effects of activin-A on FSH secretion and on the percentage of immunoreactive FSH cells were, however, not inhibited by the presence of the mitotic inhibitor cytosine arabinoside. On the other hand, the stimulatory effect of activin-A on the percentage of immunoreactive FSH cells was completely blocked in the presence of the Ca2+/calmodulin kinase inhibitor KN-62 or the L-type Ca2+ channel blocker nicardipine. Neither of these inhibitors, however, revealed significant influence on the effect of activin-A on FSH secretion. These results suggest that activin-A exhibits its dual effect on FSH cells without causing cellular proliferation. Furthermore, activin-A appears to induce increases in FSH secretion and enlargement of FSH cell population through distinct intracellular signaling pathways, the former through Ca2+-independent and the latter through Ca2+ dependent mechanisms. PMID- 11036867 TI - Stimulation of choleresis by insulin-like growth factor-I in rats. AB - Insulin-like growth factor-I (IGF-I) is an endogenous growth factor which is mainly produced in the liver. The functions of IGF-I can be summarized as growth promoting and insulin-like metabolic actions. In the present study, the effect of IGF-I on bile flow and bile acid secretion was investigated in rats. In normal rats bile flow was significantly increased by single exogenous administration of IGF-I, and by 1 week treatment of IGF-I, both bile flow and bile acid secretion were significantly increased. Moreover, to further understand the relationship between IGF-I and bile acid secretion, hypophysectomized rats were next used. We found that the decreases in bile flow and bile acid secretion observed in rats after hypophysectomy, as well as the decrease in the endogenous level of IGF-I in the blood, were partially reversed by 1 week exogenous IGF-I treatment. Overall, this study showed that IGF-I stimulates choleresis associated with an elevation of bile acid secretion in both normal and hypophysectomized rats when exogenously administered, suggesting the importance of IGF-I in the stimulation of choleresis in vivo. PMID- 11036868 TI - A struma ovarii with increased serum basement membrane components: a case report. AB - We report a case of struma ovarii with hyperthyroidism and elevated serum concentrations of type IV collagen and laminin. Circulating levels of type IV collagen and laminin were measured using specific radioimmunoassays (RIAs) for 7S collagen and the P-1 fragment of laminin, and the basement membrane components in the tumor were investigated by immunohistochemical analysis. Strong immunohistochemical staining specific for type IV collagen and for laminin was observed to be localized in the follicular walls. The serum levels of these antigens, as determined by RIA, were very high before removal of the tumor but decreased rapidly postoperatively. The present findings suggest that struma ovarii produces large amounts of type IV collagen and laminin. In addition, elevated levels of thyroid hormones might enhance the turnover of the basement membrane in various tissues. PMID- 11036869 TI - Sphingosine 1-phosphate stimulates insulin secretion in HIT-T 15 cells and mouse islets. AB - Sphingosine is involved in the regulation of cellular processes as a second messenger in various kinds of cells. Since the possible involvement of sphingosine has not been investigated in pancreatic beta-cells, we determined the expression of putative sphingosine 1-phosphate (S1P) receptors and the effect of sphingosine on pancreatic beta-cell function using a clonal Hamster beta-cell line, HIT-T 15 cells and isolated mouse islets. We showed the expression of putative S1P receptors, Edg-3 and AGR16/H218 in HIT-T 15 cells. Ten and 20 microM S1P significantly stimulated insulin secretion for 10 minutes in HIT-T 15 cells. Ten microM S1P significantly increased insulin secretion from isolated mouse islets. Ten microM S1P obviously increased intracellular Ca2+ concentration ([Ca2+]i). Fifty nM nifedipine did not affect the S1P stimulation of insulin secretion in HIT-T 15 cells. Two microM U73122 (phospholipase C inhibitor) completely deleted 10 microM S1P-induced stimulation of insulin secretion for 10 minutes, but U73343 (an inactive analogue of U73122) did not. S1P dose dependently inhibited intracellular cyclic AMP levels. Pretreatment with 100 ng/ml pertussis toxin (PTX) partially, but significantly attenuated an increase of insulin secretion by 10 microM S1P. These data suggested that PTX-sensitive G protein-dependent pathway may, at least in part, be involved in an increase of non-glucose stimulated insulin secretion by S1P through the activation of phospholipase C-Ca2+ system. PMID- 11036870 TI - Preoperative diagnosis of thyroid carcinomas by aspiration biopsy-reverse transcription-polymerase chain reaction (ABRP): a report of two cases. AB - A preoperative molecular-based diagnostic technique for thyroid papillary and anaplastic carcinomas was recently developed to detect oncofetal fibronectin (onfFN) messenger RNA (mRNA) in fine needle aspiration biopsy (FNAB) by means of reverse transcription-polymerase chain reaction (RT-PCR). We report two cases of thyroid tumors, in which cytological diagnosis was not successful but in which RT PCR analysis provided information that helped to identify the biological features of the tumors. One case could not be diagnosed by aspiration biopsy cytology (ABC) because of poor fixation; however, RT-PCR did detect the expression of onfFN mRNA, which suggested the existence of papillary carcinoma cells. After surgery, this tumor was diagnosed as a papillary carcinoma by histological examination. The other case was diagnosed as a papillary carcinoma by ABC, but onfFN mRNA was not detected in the FNAB by RT-PCR. The neoplasm was diagnosed as a follicular tumor by histological examination. These cases suggest the benefits of combining both genetic and cytological approaches to the examination of FNAB. PMID- 11036871 TI - A case of myotonic dystrophy (MD) associated with glucose-induced hyperinsulinemia followed by reactive hypoglycemia and increased number of cytosine-thymine-guanine (CTG) trinucleotide repeats in MD gene. AB - A 39-year-old man with myotonic dystrophy consulted our hospital for nausea, vomiting and dizziness that occurred after 75 g oral glucose tolerance test (OGTT). Reexamination of OGTT revealed remarkable hyperinsulinemia (622 microU/ml) followed by reactive hypoglycemia (50 mg/dl) and such hypoglycemic symptoms as nausea, vomiting, dizziness and palpitation. DNA analysis of the circulating lymphocytes revealed increased (1,500 times) number of cytosine thymine-guanine (CTG) trinucleotide repeats in myotonic dystrophy protein kinase (DM kinase) gene. Gel chromatographic analysis of the plasma in combination with sensitive enzyme immunoassay of insulin revealed that the ratio of proinsulin to total immunoreactive insulin was elevated at fasting (12.9%), and was decreased to 8.9% at 60 min after glucose administration. These findings may indicate that biologically active authentic insulin was predominantly secreted after glucose administration in the present case. This is the first case report of myotonic dystrophy with hyperinsulinemia associated with reactive hypoglycemia induced by oral glucose administration. PMID- 11036872 TI - A case of lymphocytic infundibuloneurohypophysitis showing diabetes insipidus followed by anterior hypopituitarism associated with thrombasthenia. AB - We report a case of a 42-year old male patient with diabetes insipidus followed by anterior hypopituitarism associated with thrombasthenia. The patient had been diagnosed with thrombasthenia since the age of 19. He was admitted and diagnosed as diabetes insipidus in 1995. Although T1-weighted image of magnetic resonance imaging (MRI) showed empty sella and partial pituitary stalk hypertrophy, the anterior pituitary functions were normal at that time. Three years later, he was re-admitted after an episode of general malaise and impotence in 1998. Endocrinological studies revealed adrenal insufficiency, hypothyroidism and hypogonadism. T1-weighted image of MRI demonstrated the thickening of pituitary stalk and neurohypophysis. Analysis of anti-pituitary antibodies by immunoblotting identified a major band at 61.5 kDa. The diabetes insipidus was controlled by desmopressin acetate and the shrinkage of pituitary stalk was seen after hormonal replacement therapy including glucocorticoid and thyroid hormone. We suggested that this case represented lymphocytic infundibuloneurohypophysitis, in which a chronic inflammatory process occurred in infundibulum and/or neurohypophysis and that hypopituitarism developed possibly due to damage to the pituitary portal vessels caused by a thickened pituitary stalk, although a pituitary biopsy was not done because of the risk of bleeding in thrombasthenia. The pituitary autoantibodies in sera from patients with hypopituitarism may be helpful to characterize the patient with lymphocytic hypophysitis. PMID- 11036873 TI - Interleukin (IL)-4 and IL-13 inhibit the differentiation of murine osteoblastic MC3T3-E1 cells. AB - Interleukin-4 (IL-4) inhibits the spontaneous and stimulated bone resorption resulting from the inhibition of osteoclast formation, as well as osteoclastic activity. Since IL-13 shares some biological properties with IL-4, it was recently reported that IL-13 inhibits bone resorption. The present study was designed to determine the effects of murine IL-4 (IL-4) and murine IL-13 (IL-13) on the murine osteoblastic cell line MC3T3-E1. IL-4 and IL-13 stimulated 3H thymidine incorporation in the MC3T3-E1 cells and its proliferation in dose dependent manners. A spontaneous increase in alkaline phosphatase (ALP) activity in the cells after plating was inhibited by IL-4 or IL-13, and both cytokines blunted an increase in ALP activity by human parathyroid hormone (PTH) (1-34). PTH-stimulated cyclic AMP (cAMP) production was inhibited by pretreatment with IL 4 and IL-13 for 48 hr in dose dependent manners. Pretreatment with IL-4 and IL-13 for 48 hr caused a decrease in PTH-induced cAMP production at any stimulatory concentration. However, the effective dose (ED50) was unchanged by the pretreatment with these cytokines. Pretreatment with IL-4 and IL-13 did not modulate cAMP generation by forskolin. In contrast, cAMP generation by PGE2 is greater in the cells treated with the cytokines compared to those without the cytokines. These results indicate that IL-4 and IL-13 act on MC3T3-E1 cells in the same manner, stimulating cell proliferation, but inhibiting cell differentiation. The inhibition of osteoblast differentiation by IL-4 and IL-13 may be associated with a decrease in PTH actions on osteoblasts. PMID- 11036874 TI - Multiple branchial cleft-like cysts in a female patient with Hashimoto's thyroiditis. AB - We report a case of branchial cleft-like cysts (intrathyroidal lymphoepithelial cysts) associated with Hashimoto's thyroiditis. Palpation did not detect any nodules. Multiple cystic lesions were detected in the lateral side of the thyroid bilateral lobes by imagings of an I-123 scintigram, Tl-201 scintigram, sonography, and computerized tomography. Sonography displayed multiple cysts with strong echogenic spots in the cystic fluid. Repeated fine needle aspiration biopsies of the cysts consistently revealed only normal lymphocytes. Although these lesions could not be given diagnosis, subtotal thyroidectomy leaving the intact isthmus was performed. Microscopic findings revealed multiple branchial cleft-like cysts lined by flattened epithelial cells. Surrounding the epithelial lining were dense lymphoid follicles with large, reactive germinal centers. The remaining thyroid parenchyma showed Hashimoto's thyroiditis. Multiple branchial cleft-like cysts should be considered when sonographic examination reveals multiple cysts in the lateral side of the bilateral lobes, and fine needle aspiration biopsy displays only normal lymphocytes. To our knowledge, this is the first case of branchial cleft-like cysts associated with Hashimoto's thyroiditis reported in Japan. PMID- 11036875 TI - Characterization of an early decline in baseline plasma glucose concentration after acute insulin elevation during euglycemic hyperinsulinemic clamp in patients with type 2 diabetes mellitus. AB - To investigate the contribution of the liver to whole-body insulin resistance in patients with type 2 diabetes mellitus, we analyzed the early decline (slope "a") in the baseline plasma glucose level following acute hyperinsulinemia in the initial phase of a euglycemic hyperinsulinemic clamp study, rather than using an isotope-dilution method. Slope "a" was comparable among groups of diabetic and non-diabetic subjects, and did not correlate well with glucose infusion rate (GIR), an index of peripheral (primarily skeletal muscle) insulin resistance. In contrast, slope "a" was significantly lower in obese (BMI > 25) type 2 diabetic patients compared with their non-obese counterparts, consistent with the general belief that obesity is a condition of insulin resistance in liver as well as in peripheral tissues. A subset of six insulin-resistant (nearly zero GIR) type 2 diabetic patients (pubertal adolescents) demonstrated a markedly blunted slope "a". Their insulin resistance (GIR) substantially recovered concomitant with an increase in slope "a" after pretreatment with somatostatin analogue in two cases studied, suggesting possible suppression of hepatic glucose production through lowering of plasma glucagon concentrations. Furthermore, slope "a" correlated significantly (r = -0.480, p<0.0001) with HOMA index (FPG x FIRI), the latter being recently regarded as an index of hepatic insulin resistance. These data showed that slope "a" obtained from euglycemic hyperinsulinemic clamp may be a clinically useful index of hepatic insulin resistance rather than an index of peripheral insulin resistance. PMID- 11036876 TI - Changes in proliferative potential, apoptosis and Bcl-2 protein expression in cytotrophoblasts and syncytiotrophoblast in human placenta over the course of pregnancy. AB - In order to evaluate placental trophoblast proliferation and apoptosis during pregnancy, we investigated proliferating cell nuclear antigen (PCNA) expression, apoptosis and Bcl-2 protein expression in the human placenta using avidin/biotin immunoperoxidase method to examine PCNA and Bcl-2 protein expression, and TUNEL method to assess apoptosis. The appearance of apoptotic cells in very early term placental trophoblasts was also examined by transmission electron microscopy. PCNA was immunolocalized in the nuclei of cytotrophoblasts (C-cells). Determination of the mean percentage of PCNA-positive nuclei of C-cells revealed that PCNA expression in C-cells was highest in very early term (4th to 5th wk) placentas and significantly decreased with the advance of pregnancy. Bcl-2 protein was immunolocalized in the cytoplasm of syncytiotrophoblast (S-cell), being least abundant in very early term placentas, less abundant in early term and midterm placentas, and most abundant in term placentas. On the basis of TUNEL method, apoptosis was apparent in the nuclei of both C-cells and S-cell. The apoptosis positive rate of C-cell nuclei was highest in very early term 4th to 5th wk placentas, and significantly decreased in early term 7th to 9th wk and midterm placentas, but somewhat increased in term placentas compared to that in midterm placentas. On the other hand, apoptosis positive rate of S-cell nuclei was remarkably higher only in very early term 4th to 5th wk placentas compared to that in early term, midterm and term placentas. Transmission electron microscopy revealed the appearance of apoptotic nucleus in very early term placental trophoblasts. These results demonstrate for the first time that apoptosis in the human normal placenta predominates in both C-cells and S-cell in very early term 4th to 5th wk pregnancy and drastically diminished after 7th wk of pregnancy. An apparent increase in apoptosis in C-cells in term placentas compared to that in midterm placentas may reflect aging of the placenta or parturition-associated biological change. The abundant expression of Bcl-2 protein in S-cell in term placentas may be responsible for the diminished occurrence of apoptosis in S-cell in term placentas. PMID- 11036877 TI - Apoplexy of pituitary macroadenoma after combined test of anterior pituitary function. AB - Pituitary apoplexy has been reported as a very rare complication of combined tests of anterior pituitary function and of TRH or gonadotropin-releasing hormone (GnRH) administration in pituitary tumor. A 34-year-old man with a GH-secreting pituitary macroadenoma and diabetes mellitus received an injection of 400 microg TRH, 100 microg GnRH, and 0.15 U/Kg regular insulin. Twenty minutes later, he complained of a severe headache and vomited. Visual acuity and visual field did not change and his headache was persistent during the next 24 hours of conservative management. Magnetic resonance imaging (MRI) of the sella turcica done the day after the event showed definitive elevation of the optic chiasm and slight enlargement of tumor and focal areas of mixed high signal and low signal intensities in the macroadenoma on noncontrast T1-weighted images. Headache subsided markedly within a day of octreotide therapy. Transsphenoidal removal of the pituitary tumor was performed 9 days after the hormone study. Ischemic necrosis and hemorrhage were confirmed in the acidophilic adenoma with positive immunostaining for GH. Postoperative course was uneventful and his serum insulin like growth factor-1 (IGF-1) level and blood glucose levels were normalized. Three months after the surgery the dynamic test was repeated without adverse effects. To our knowledge, this is a very rare case of apoplexy of GH-secreting pituitary adenoma after a combined stimulation test of anterior pituitary function. PMID- 11036878 TI - Multifocal fibrosclerosis as a possible cause of panhypopituitarism with central diabetes insipidus. AB - Multifocal fibrosclerosis denotes a combination of similar fibrous disorders occurring at different anatomical sites. We encountered a 53-year-old male patient with orbital pseudotumor, chronic paranasal sinusitis, fibrous nodules of the lungs, intracranial pachymeningitis, and panhypopituitarism with central diabetes insipidus (DI) as a possible manifestation of multifocal fibrosclerosis. It has been reported that intracranial pachymeningitis or orbital pseudotumor associated with multifocal fibrosclerosis could invade the sella turcica causing a variety of anterior and/or posterior pituitary dysfunctions. In our case, intracranial pachymeningitis apparently involved the pituitary stalk and gland. Isolated gonadotropin deficiency, in addition to central DI, preceded panhypopituitarism. Although panhypopituitarism with central DI due to multifocal fibrosclerosis is quite rare and only one case has ever been reported, this systemic fibrotic disorder can be a possible cause of panhypopituitarism with central DI. PMID- 11036879 TI - Possible reproductive toxicity of styrene in peripubertal male mice. AB - Environmental estrogens (endocrine disruptive chemicals) have been shown to affect reproduction in wild life and it has been reported that maternal exposure to those chemicals has adverse effects on the male reproductive tract. However, little is known about the potential effects of prepubertal or pubertal exposure to environmental estrogens on the male reproductive tract. Here we examine plasma hormone levels of mice following 4-week oral administration of styrene. Plasma free testosterone levels were dramatically decreased following 4 weeks of styrene treatment compared with control group. No differences in plasma corticosterone and luteinizing hormone levels were seen between styrene and control groups. Thus, exposure to styrene around pubertal period may directly disrupt the male reproductive tract. These facts suggest that more detailed studies regarding assessment of the risk to the developing human testes from exposure to styrene and other environmental estrogens in prepubertal and pubertal period are warranted. PMID- 11036880 TI - Thyroxine prophylaxis after bilateral subtotal thyroidectomy for multinodular goiter. AB - In this study, we investigated the value of thyroxine administration to prevent recurrence after bilateral subtotal thyroidectomy for multinodular goiter. Patients that had benign multinodular goiter were operated on with the same surgical principles: ligation of both superior and inferior thyroid arteries on both sides, bilateral subtotal resection of thyroid gland including all visible nodules. On the 3rd postoperative day, the patients were divided into two groups: with 100 microgram 1-thyroxine daily (Thyroxine group) or no therapy (Control group). No recurrences were encountered among 40 patients followed up for 6 months and 20 patients for at least one year. One patient in the control group developed manifest hypothyroidism (5.3%). The mean TSH level of the control group was significantly higher than that of thyroxine group at 1st, 2nd, 3rd, 4th, 5th, 6th, and 12th months. At the end of the first year, the mean TSH level of the control group was four times that of the normal. On the other hand, the mean TSH level of the thyroxine group was within normal limits but not suppressed. In conclusion, we found that the pituitary-thyroid axis did not become normal spontaneously one year after thyroidectomy. Therefore, postoperative thyroxine administration seems to be of value, especially in endemic regions like Turkey. PMID- 11036881 TI - Altered CD15 glycolipid expression in the developing rat cerebellum following treatment with antithyroid drug, propylthiouracil. AB - Thyroid hormone plays an important role during central nervous system (CNS) development. Experimentally-induced perinatal hypothyroid rats show abnormal cerebellar cytoarchitecture, but the underlying mechanism is not fully understood. Altered cell-cell interactions may contribute to such abnormalities, since the expression of NCAM is increased in hypothyroid animals. In the present study, we examined the expression of carbohydrate epitope 3-fucosyl-N-acetyl lactosamine (CD15 antigen), that can be localized on both astrocytes and neurons in the developing brain, and is considered to play an important role in glial neuronal interaction and cell migration. Newborn rats were treated with an antithyroid drug, propylthiouracil (PTU) and the CD15 glycolipid levels in the cerebellum were examined by enzyme-linked immunosolvent assay (ELISA) using 7A monoclonal antibody raised against rat forebrain antigen. A transient elevation of CD15 level was observed on postnatal day 10 in PTU-treated animals. Analysis of neutral glycolipids on high performance thin layer chromatography (HPTLC), revealed two distinct immunoreactive bands, corresponding to Fuc-nLc6 and Fuc nLc4. The Fuc-nLc4 is preferentially increased in the PTU-treated group. These results suggest that a transient increase in CD15 glycoconjugates with isoform specific manner induced by PTU may contribute to morphological abnormalities in hypothyroid rat cerebellum affecting granule cell migration. PMID- 11036882 TI - Crucial role of insulin in leptin maintenance: profound decrease in serum leptin by octreotide acetate in insulinoma subjects. AB - To further clarify the relationship between insulin and leptin, time course changes in plasma glucose, serum insulin and leptin levels were analyzed after subcutaneous administration of 100 microg octreotide acetate in two insulinoma subjects. Octreotide acetate induced a prompt decrease in serum insulin level, accompanied with an increase in plasma glucose in both patients. Following the decrease in serum insulin level, serum leptin concentrations were profoundly decreased by 66% and 44%, 8-12 hrs after octreotide injection; that is, the concentrations decreased from 41.1 to 13.8 ng/ml in patient 1, and from 17.5 to 9.8 ng/ml in patient 2. Daily profiles of plasma glucose, serum insulin and leptin without octreotide administration did not show such alterations in these indexes in patient 1. These data show that circulating leptin may be susceptible to decline dependent on the decrease in serum insulin, suggesting that insulin plays a crucial role in the maintenance of leptin secretion in humans. PMID- 11036883 TI - Genomic DNA analysis of thyrotropin receptor in a family with hereditary hyperthyroidism. AB - Mutations of the thyrotropin receptor (TSH-R) gene have been reported in some cases of hyperthyroidism. We report a case of a family that had a high incidence of hyperthyroidism (6/13) which strongly suggested hereditary factors. We then analyzed whether the family had mutations of the TSH-R gene. No significant mutations in exon 10 of the TSH-R gene were found in the patient by restriction fragment length polymorphism analysis and polymerase chain reaction direct sequencing, when compared with those with 4 normal subjects and 2 patients with Graves' disease. Unknown mutations in the extracellular region of the receptor or other genes in this family remain to be studied. PMID- 11036884 TI - The heart in context. PMID- 11036885 TI - One step forward or one step back with tamoxifen? PMID- 11036886 TI - A late step in the right direction of stroke care. PMID- 11036887 TI - Progressive supranuclear palsy in the molecular age. PMID- 11036888 TI - How strong and how wide is the link between HPV and oropharyngeal cancer? PMID- 11036889 TI - Ureteric stents, far from ideal. PMID- 11036890 TI - Towards invasive diagnostic techniques as standard management of ventilator associated pneumonia. PMID- 11036891 TI - Serious life events and congenital malformations: a national study with complete follow-up. AB - BACKGROUND: Emotional stress during organogenesis could, in theory, cause congenital malformations. We undertook a follow-up study to test the hypothesis that psychosocial stress increases the prevalence of malformations, particularly of the cranial neural crest. METHODS: We defined severe life events as death or first hospital admission for cancer or acute myocardial infarction in partners or children. From 1980 to 1992 all women exposed to severe life events during pregnancy and up to 16 months previously were identified by means of five national registers. We included 3560 exposed pregnancies and 20,299 pregnancies without such exposures randomly selected as a control cohort. FINDINGS: The frequency of cranial-neural-crest malformations was higher in pregnancies with exposure to severe life events than in those without such exposure (42 [1.18%] vs 131 [0.65%]; adjusted odds ratio 1.54 [95% CI 1.05-2.27]). For other malformations, the frequencies were 3.04% and 3.26% (1.14 [0.94-1.42]). Women exposed in two consecutive pregnancies had a higher risk of cranial-neural-crest malformations (2.99 [1.06-8.43]). Death of an older child during the first trimester was associated with an adjusted odds ratio of cranial-neural-crest malformations of 4.75 (1.63-13.8). Unexpected death of a child during the first trimester was associated with adjusted odds ratios of 8.36 (2.41-29.0) for cranial-neural-crest malformations and 3.64 (1.29-10.3) for other malformations. INTERPRETATION: These findings support the hypothesis that severe emotional stress during pregnancy, especially that related to death of a child, may cause congenital malformations, particularly those of the cranial neural crest. PMID- 11036892 TI - Risk and prognosis of endometrial cancer after tamoxifen for breast cancer. Comprehensive Cancer Centres' ALERT Group. Assessment of Liver and Endometrial cancer Risk following Tamoxifen. AB - BACKGROUND: Tamoxifen increases the risk of endometrial cancer. However, few studies have produced reliable risk estimates by duration, dose, and recency of use, or addressed the prognosis of endometrial cancers in tamoxifen-treated women. METHODS: We did a nationwide case-control study on the risk and prognosis of endometrial cancer after tamoxifen use for breast cancer. Information on tamoxifen use and other risk factors for endometrial cancer was obtained from 309 women with endometrial cancer after breast cancer (cases), and 860 matched controls with breast cancer but without endometrial cancer. For 276 cases, we obtained tissue blocks of endometrial cancer to review the diagnosis, and used immunohistochemistry to examine hormone-receptor status and overexpression of p53. FINDINGS: Tamoxifen had been used by 108 (36.1%) of 299 cases and 245 (28.5%) controls (relative risk 1.5 [95% CI 1.1-2.0]). Risk of endometrial cancer increased with longer duration of tamoxifen use (p < 0.001), with relative risks of 2.0 (1.2-3.2) for 2-5 years and 6.9 (2.4-19.4) for at least 5 years compared with non-users. Endometrial cancers of stage III and IV occurred more frequently in long-term tamoxifen users (> or = 2 years) than in non-users (17.4% vs 5.4%, p=0.006). Long-term users were more likely than non-users to have had malignant mixed mesodermal tumours or sarcomas of the endometrium (15.4% vs 2.9%, p < or = 0.02), p53-positive tumours (31.4% vs 18.2%, p=0.05), and negative oestrogen receptor concentrations (60.8% vs 26.2%, p < or = 0.001). 3-year endometrial cancer-specific survival was significantly worse for long-term tamoxifen users than for non-users (76% for > or = 5 years, 85% for 2-5 years vs 94% for non users, p=0.02). INTERPRETATION: Long-term tamoxifen users have a worse prognosis of endometrial cancers, which seems to be due to less favourable histology and higher stage. However, the benefit of tamoxifen on breast-cancer survival far outweighs the increased mortality from endometrial cancer. Nevertheless, we seriously question widespread use of tamoxifen as a preventive agent against breast cancer in healthy women. PMID- 11036893 TI - A palliative-care intervention and death at home: a cluster randomised trial. AB - BACKGROUND: The Palliative Medicine Unit at University Hospital of Trondheim, Norway, started an intervention programme that aims to enable patients to spend more time at home and die there if they prefer. Close cooperation was needed with the community health-care professionals, who acted as the principal formal caregivers, and a multidisciplinary consultant team coordinated the care. We did a cluster randomised trial to assess the intervention's effectiveness compared with conventional care METHODS: Community health-care districts in and around Trondheim, Norway, were defined as the clusters to be randomised. We enrolled 434 patients (235 assigned intervention and 199 conventional care [controls]) in these districts who had incurable malignant disease and an expected survival of 2 9 months. Main outcomes were place of death and time spent in institutions in the last month of life. FINDINGS: 395 patients died. Of these, more intervention patients than controls died at home (54 [25%] vs 26 [15%], p<0.05). The time spent at home was not significantly increased, although intervention patients spent a smaller proportion of time in nursing homes in the last month of life than did controls (7.2 vs 14.6%, p<0.05). Hospital use was similar in the two groups. INTERPRETATION: The palliative-care intervention enabled more patients to die at home. More resources for care in the home (palliative care training and staff) and an increased focus on use of nursing homes would be necessary, however, to increase time at home and reduce hospital admissions. PMID- 11036894 TI - Alternative strategies for stroke care: a prospective randomised controlled trial. AB - BACKGROUND: Organised specialist care for stroke improves outcome, but the merits of different methods of organisation are in doubt. This study compares the efficacy of stroke unit with stroke team or domiciliary care. METHODS: A single blind, randomised, controlled trial was undertaken in 457 acute-stroke patients (average age 76 years, 48% women) randomly assigned to stroke unit, general wards with stroke team support, or domiciliary stroke care, within 72 h of stroke onset. Outcome was assessed at 3, 6, and 12 months. The primary outcome measure was death or institutionalisation at 12 months. Analyses were by intention to treat. FINDINGS: 152 patients were allocated to the stroke unit, 152 to stroke team, and 153 to domiciliary stroke care. 51 (34%) patients in the domiciliary group were admitted to hospital after randomisation. Mortality or institutionalisation at 1 year were lower in patients on a stroke unit than for those receiving care from a stroke team (21/152 [14%] vs 45/149 [30%]; p<0.001) or domiciliary care (21/152 [14%] vs 34/144 [24%]; p=0.03), mainly as a result of reduction in mortality. The proportion of patients alive without severe disability at 1 year was also significantly higher on the stroke unit compared with stroke team (129/152 [85%] vs 99/149 [66%]; p<0.001) or domiciliary care (129/152 [85%] vs 102/144 [71%]; p=0.002). These differences were present at 3 and 6 months after stroke. INTERPRETATION: Stroke units are more effective than a specialist stroke team or specialist domiciliary care in reducing mortality, institutionalisation, and dependence after stroke. PMID- 11036895 TI - First permanent implant of the Jarvik 2000 Heart. AB - BACKGROUND: Heart failure is a major public-health concern. Quality and duration of life on maximum medical therapy are poor. The availability of donor hearts is severely limited, therefore an alternative approach is necessary. We have explored the use of a new type of left-ventricular assist device intended as a long-term solution to end-stage heart failure. METHODS: As part of a prospective clinical trial, we implanted the first permanent Jarvik 2000 Heart--an intraventricular device with an innovative power delivery system--into a 61-year old man (New York Heart Association functional class IV) with dilated cardiomyopathy. We assessed the effect of this left-ventricular assist device on both native heart function and the symptoms and systemic characteristics of heart failure. FINDINGS: The Jarvik 2000 Heart sustained the patient's circulation, and was practical and user-friendly. After 6 weeks, exercise tolerance, myocardial function, and end-organ function improved. Symptoms of heart failure have resolved, and continuous decreased pulse-pressure perfusion has had no adverse effects in the short term. There has been no significant haemolysis and no device related complications. The skull-mounted pedestal is unobtrusive and has healed well. CONCLUSIONS: The initial success of this procedure raises the possibility of a new treatment for end-stage heart failure. In the longer term, its role will be determined by mechanical reliability. PMID- 11036896 TI - Mother-to-child transmission of hepatitis C virus: evidence for preventable peripartum transmission. AB - BACKGROUND: Little information is available about the timing of mother-to-child transmission of hepatitis C virus (HCV), and no interventions to decrease transmission rates have been identified. We examined the effect of risk factors, including mode of delivery, on the vertical transmission rate. METHODS: Data from HCV-infected women and their infants from three hospitals in Ireland and from a British Paediatric Surveillance Unit study of infants born to HCV-infected mothers were used to estimate the vertical transmission rate and risk factors for transmission. We used a probabilistic model using methods that simultaneously estimated the time to HCV-antibody loss in uninfected infants and the diagnostic accuracy of PCR tests for HCV RNA. FINDINGS: 441 mother-child pairs from the UK (227) and Ireland (214) were included. 50% of uninfected children became HCV antibody negative by 8 months and 95% by 13 months. The estimated specificity of PCR for HCV RNA was 97% (95% CI 96-99) and was unrelated to age; sensitivity was only 22% (7-46) in the first month but rose sharply to 97% (85-100) thereafter. The vertical transmission rate was 6.7% (4.1-10.2) overall, and 3.8 times higher in HIV coinfected (n=22) than in HIV-negative women after adjustment for other factors (p=0.06). No effect of breastfeeding on transmission was observed, although only 59 women breastfed. However, delivery by elective caesarean section before membrane rupture was associated with a lower transmission risk than vaginal or emergency caesarean-section delivery (odds ratio 0 [0-0.87], p=0.04, after adjustment for other factors). INTERPRETATION: The low sensitivity of HCV RNA soon after birth and the finding of a lower transmission rate after delivery by elective caesarean section suggest that HCV transmission occurs predominantly around the time of delivery. If the findings on elective caesarean section are confirmed in other studies, the case for antenatal HCV testing should be reconsidered. PMID- 11036897 TI - A contrast crisis. PMID- 11036898 TI - Combined imaging techniques for pancreatic cancer. AB - Advanced pancreatic cancer has a poor prognosis. Early detection with imaging techniques can, however, improve the outlook for patients who undergo surgical resection, the only potential curative treatment. Individual techniques, however, have poor sensitivity for small masses and cannot easily differentiate tumour tissue from pancreatic masses associated with chronic pancreatitis. We combined biochemical detection on positron emission tomography with the anatomical accuracy of computed tomography and were able to improve accuracy of interpretation of imaging for patients with pancreatic cancer. PMID- 11036899 TI - Corticosteroid-responsive diabetes mellitus associated with autoimmune pancreatitis. AB - Autoimmune pancreatitis, which can be treated with corticosteroid therapy, has the potential to induce diabetes. In a cohort study of 2220 patients with suspected chronic pancreatitis, we found that all four patients with autoimmune chronic pancreatitis also had diabetes. Treatment with prednisolone subsequently improved insulin secretion and glycaemic control in these patients. PMID- 11036900 TI - Pesticide exposure and risk of mild cognitive dysfunction. AB - Little is known about the adverse effects of substances, such as pesticides and metals, on the development of mild cognitive dysfunction (MCD). Cross-sectional and prospective data from the Maastricht Aging Study were used to find out the potential neurotoxicity of particular substances. Exposure to pesticides, for example by arable farmers and gardeners, was associated with increased risks of MCD. Exposure to metals and organic solvents was not associated with MCD. Our findings might reflect subtle changes in brain function among people exposed to pesticides. PMID- 11036901 TI - Increased prevalence of renal disease in silica-exposed workers. AB - We report an increased prevalence of renal disease, particularly glomerulonephritis, in Upper Weardale, UK. Silica exposure during fluorspar mining and processing is a likely cause. PMID- 11036902 TI - Association between mutations of the follicle-stimulating-hormone receptor and repeated twinning. AB - Follicle-stimulating hormone (FSH) has a role in folliculogenesis and spontaneous twinning. Using the candidate gene approach, we searched for mutations in the gene encoding the FSH receptor in a woman who had given birth to two sets of dizygotic twins without fertility treatment. We identified two linked mutations (Thr307Ala and Asn680Ser) that were closely associated with this phenotype. We suggest that expression of both mutations increases the sensitivity of the receptor to FSH. PMID- 11036903 TI - Putting nerve cell development into reverse. PMID- 11036904 TI - Pseudomonas genome reveals a formidable foe. PMID- 11036905 TI - Row continues over proposed reform of residential care in UK. PMID- 11036906 TI - International concern grows over psychiatric abuses in China. PMID- 11036907 TI - Austria investigates allegations of tainted-blood exports. PMID- 11036908 TI - WHO draws attention to women's health in south-east Asia. PMID- 11036909 TI - Traumatic brain injury. AB - The decrease in mortality and improved outcome for patients with severe traumatic brain injury over the past 25 years can be attributed to the approach of "squeezing oxygenated blood through a swollen brain". Quantification of cerebral perfusion by monitoring of intracranial pressure and treatment of cerebral hypoperfusion decrease secondary injury. Before the patient reaches hospital, an organised trauma system that allows rapid resuscitation and transport directly to an experienced trauma centre significantly lowers mortality and morbidity. Only the education of medical personnel and the institution of trauma hospital systems can achieve further improvements in outcome for patients with traumatic brain injuries. PMID- 11036910 TI - The endotoxin-lipoprotein hypothesis. AB - The advent of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) has revolutionised the treatment of hypercholesterolaemia. Statin treatment, by lowering the atherogenic lipoprotein profile, reduces morbidity and mortality in patients with cardiovascular disease. Treatment with simvastatin causes a reduction of events of new-onset heart failure, but this may be attributable to properties other than its lipid-lowering effects. There is some evidence that lower serum cholesterol concentrations (as a surrogate for the totality of lipoproteins) relate to impaired survival in patients with chronic heart failure (CHF). Inflammation is a feature in patients with CHF and increased lipopolysaccharide may contribute substantially. We postulate that higher concentrations of total cholesterol are beneficial in these patients. This is potentially attributable to the property of lipoproteins to bind lipopolysaccharide, thereby preventing its detrimental effects. We hypothesise there is an optimum lipoprotein concentration below which lipid reduction would, on balance, be detrimental. We also propose that, in patients with CHF, a non lipid-lowering statin (with ancillary properties such as immune modulatory and anti-inflammatory actions) could be as effective or even more beneficial than a lipid-lowering statin. PMID- 11036911 TI - The idea is more important than the experiment. PMID- 11036912 TI - Adult cardiovascular risk factors in premature babies. PMID- 11036913 TI - Adult cardiovascular risk factors in premature babies. PMID- 11036914 TI - Adult cardiovascular risk factors in premature babies. PMID- 11036916 TI - Gelastic epilepsy caused by hypothalamic hamartoma. PMID- 11036915 TI - Adult cardiovascular risk factors in premature babies. PMID- 11036917 TI - Short-acting beta2-agonists. PMID- 11036918 TI - Mitochondrial DNA recombination. PMID- 11036919 TI - Early non-invasive management of acute coronary syndromes. PMID- 11036920 TI - Depressive state and common cold. PMID- 11036921 TI - Selenium and human health. PMID- 11036922 TI - Selenium and human health. PMID- 11036923 TI - Selenium and human health. PMID- 11036924 TI - Selenium and human health. PMID- 11036925 TI - False negatives in oestrogen-receptor assay. PMID- 11036926 TI - Mother-to-child HIV-1 transmission. PMID- 11036927 TI - End-of-life decisions for newborn infants. PMID- 11036928 TI - End-of-life decisions for newborn infants. PMID- 11036929 TI - Irregular gout. PMID- 11036930 TI - Multistep signaling and transcriptional requirements for pituitary organogenesis in vivo. AB - During development of the mammalian pituitary gland, specific hormone-producing cell types, critical in maintaining homeostasis, emerge in a spatially and temporally specific fashion from an ectodermal primordium. We have investigated the molecular basis of generating diverse cell phenotypes from a common precursor, providing in vivo and in vitro evidence that development of these cell types involves at least four sequential phases of signaling events and the action of a gradient at an ectodermal boundary. In the first phase, we hypothesize that this notochord induces invagination of Rathke's pouch from the oral ectoderm. This is followed by appearance of an ectodermal boundary, formed with exclusion of Shh from the nascent pouch. Next, signals from the ventral diencephalon- expressing BMP4, Wnt5a, FGF10, and FGF8--in concert with Shh represent critical in vivo signals for pituitary determination. Subsequently, a dorsal-ventral BMP2 signal gradient emanates from a ventral pituitary organizing center, forming at the boundary to oral ectoderm region from which Shh expression is selectively excluded. In concert with a dorsal FGF8 signal, this creates opposing gradients that generate overlapping patterns of specific transcription factors that underlie cell lineage specification events. The mechanisms by which these transient gradients of signaling molecules lead to the appearance of four ventral pituitary cell types appear to involve the reciprocal interactions of two transcription factors, Pit-1 and GATA-2, which are epistatic to the remainder of the cell type-specific transcription programs and serve as a molecular memory of the transient signaling events. Unexpectedly, this program includes a DNA-binding independent function of Pit-1, suppressing the ventral GATA-2-dependent gonadotrope program by inhibiting GATA-2 binding to gonadotrope- but not thyrotrope-specific genes. This indicates that both DNA-binding-dependent and independent actions of abundant determining factors contribute to generate distinct cell phenotypes. In the fourth phase, temporally specific loss of the BMP2 signal is required to allow terminal differentiation. The consequence of these sequential organ and cellular determination events is that each of the pituitary cell types--gonadotropes, thyrotropes, somatotropes, lactotropes, corticotropes, and melanotropes appears to be determined, in a ventral to dorsal gradient, respectively, apparently based on a combinatorial code of transcription factors induced by the gradient of specific signaling molecules. PMID- 11036931 TI - Vascular endothelial growth factor and the regulation of angiogenesis. AB - The development of a vascular supply is essential not only for organ development and differentiation during embryogenesis but also for wound healing and reproductive functions in the adult Folkman, 1995). Angiogenesis is also implicated in the pathogenesis of a variety of disorders: proliferative retinopathies, age-related macular degeneration, tumors, rheumatoid arthritis, and psoriasis (Folkman, 1995; Garner, 1994). Several potential regulators of angiogenesis have been identified, including fibroblast growth factor-a (aFGF), bFGF, transforming growth factor-alpha (TGF-alpha), TGF-beta, hepatocyte growth factor/scatter factor (HGF/SF), tumor necrosis factor-alpha (TNF-alpha), angiogenin, and interleukin-8 (IL-8) (Folkman and Shing, 1992; Risau, 1997). More recently, the angiopoietins, the ligands of the Tie-2 receptor (Suri et al., 1996; Maisonpierre et al., 1997), have been identified. Vascular endothelial growth factor (VEGF) is an endothelial-cell-specific mitogen. The finding that VEGF was potent and specific for vascular endothelial cells and, unlike bFGF, freely diffusible, led to the hypothesis that this molecule plays a unique role in the regulation of physiological and pathological angiogenesis (Ferrara and Henzel, 1989: Leung et al., 1989). Over the last few years, several additional members of the VEGF gene family have been identified, including placenta growth factor (PIGF) (Maglione et al., 1991,1993), VEGF-B (Olofsson et al., 1996), VEGF C (Joukov et al., 1996; Lee et al., 1996), and VEGF-D (Orlandini et al., 1996. Achen et al., 1998). There is compelling evidence that VEGF plays an essential role in the development and differentiation of the cardiovascular system (Ferrara and Davis-Smyth, 1997). PMID- 11036932 TI - Placental prolactins and the physiology of pregnancy. AB - Mammalian pregnancy is characterized by a concerted and widespread series of changes in maternal physiology, many of which are direct responses to the binding of placental hormones to maternal targets. Among these placental hormones are proteins closely related to prolactin. In rodents, a large number of these placental prolactin-related hormones are expressed that have a broad spectrum of activities, including activities on endothelial cells and blood cells. PMID- 11036933 TI - Autocrine and paracrine Mullerian inhibiting substance hormone signaling in reproduction. AB - Members of the transforming growth factor beta (TGFbeta) superfamily are polypeptide growth factors that exhibit diverse effects on normal cell growth, adhesion, mesenchymal-epithelial interactions, cell differentiation, and programmed cell death. This chapter will discuss the work of ourselves and others on one member of this large superfamily, Mullerian inhibiting substance (MIS, or anti-Mullerian hormone, AMH) and its role in reproductive tract development and the adult gonad. Using recombinant MIS protein, it is possible to begin unraveling the molecular mechanism of duct involution in the embryo. Our recent results suggest that MIS triggers cell death by altering mesenchymal-epithelial interactions. In addition to the developmental effects of MIS in secondary sexual differentiation, expression studies of the MIS ligand and the MIS type II receptor (MISIIR) suggest a potential regulatory role for MIS in adult germ cell maturation and gonadal function. Recent data from others suggest that MIS may act in a paracrine manner to block differentiation of interstitial cells of the adult gonad by repressing all or some steps of steroidogenesis. Our studies are highly suggestive of direct repression of steroidogenic enzyme gene expression by activation of the MIS signaling pathway. Thus, for the first time, an opportunity to define fully target genes and components of the MIS signaling pathway may be possible. PMID- 11036934 TI - Chronic hypersecretion of luteinizing hormone in transgenic mice disrupts both ovarian and pituitary function, with some effects modified by the genetic background. AB - When the pituitary or hypothalamus becomes resistant to steroid negative feedback, a vicious cycle ensues, resulting in chronic hypersecretion of luteinizing hormone (LH) from the pituitary and steroids from the ovaries. In women, LH hypersecretion is implicated in infertility, miscarriages, and development of granulosa cell tumors. Progress in defining the underlying mechanisms of LH toxicity, however, has been limited by the lack of well-defined animal models. To that end, we have developed a new transgenic mouse model (alpha LHbetaCTP) wherein LH hypersecretion occurs chronically and results in several dire pathological outcomes. Chronic hypersecretion of LH was achieved by introducing a transgene containing a bovine alpha subunit promoter fused to the coding region of a chimeric LHbeta subunit. The alpha subunit promoter directs transgene expression only to gonadotropes. The LHbeta chimera contains the carboxyl-terminal peptide (CTP) of the human chorionic gonadotropin beta subunit linked to the carboxyl terminus of bovine LHbeta. This carboxyl extension extends the half-life of LH heterodimers that contain the chimeric beta subunit. In intact alpha-LHbetaCTP females, serum LH is elevated five- to ten-fold in comparison to nontransgenic littermates. Levels of testosterone (T) and estradiol (E2) also are elevated, with an overall increase in the T-to-E2 ratio. These transgenic females enter puberty precociously but are anovulatory and display a prolonged luteal phase. Anovulation reflects the absence of gonadotropin releasing hormone (GnRH) and the inability to produce a pre-ovulatory surge of LH. The ovaries are enlarged, with reduced numbers of primordial follicles and numerous, giant, hemorrhagic follicles. Despite the pathological appearance of the ovary, females can be superovulated and mated. Although pregnancy occurs, implantation is compromised due to defects in uterine receptivity. In addition, pregnancy fails at midgestation, reflecting a maternal defect presumably due to estrogen toxicity. When the transgene is in a CF-1 background, all females develop granulosa cell tumors and pituitary hyperplasia by five months of age. They die shortly thereafter due to bladder atony and subsequent kidney failure. When the transgene is placed in other strains of mice, their ovaries develop a luteoma rather than a granulosa cell tumor and the pituitary develops pituitary hyperplasia followed by adenoma. In summary, alpha-LHbetaCTP mice provide a direct association between abnormal secretion of LH and development of a number of ovarian and pituitary pathological responses. PMID- 11036935 TI - Apolipoprotein B: from editosome to proteasome. AB - Apolipoprotein (apo) B, the protein component of low-density lipoproteins (LDLs), has been under intense investigation for the last three decades. During the first decade after its initial description, most reports dealt with the physical chemical characterization of apoB in its natural environment (i.e., intact LDL particles). A few studies dealing with attempts to elucidate the primary structure of apoB were published at this time (Deutsch et al., 1978; Bradley et al., 1980). However, most of these, in retrospect, represented heroic efforts that were doomed to failure because of the huge size and insoluble nature of apoB, once it is separated from its lipid environment. Indeed, during the 1970s, there was no universal agreement on the true molecular weight of the protein, which was not established until sometime into the second decade of apoB research (Yang et al., 1986b). The next 10 years were punctuated by breakthroughs on three different fronts in our understanding of apoB. The first exciting discovery was that apoB exists in two forms, apoB-100 and apoB-48 (Kane et al., 1980; Elovson et al., 1981). The next breakthrough was the elucidation of the primary structure of apoB-100 by a combination of cDNA cloning (Chen et al., 1986; Knott et al., 1986; Yang et al., 1986a) and direct peptide sequencing (Yang et al., 1986a, 1989). This decade of renaissance in apoB research was concluded by the elucidation of the structure of apoB-48. More important in terms of basic cellular molecular biology was the discovery of RNA editing, when apoB-48 was found to be the translation product of an edited apoB mRNA (Chen et al., 1987; Powell et al., 1987). RNA editing had just been described for a kinetoplastid protozoa the year before (Benne et al., 1986). ApoB mRNA editing was the first instance of RNA editing described in a higher eukaryote (Chan and Seeburg, 1995; Grosjean and Benne. 1998). The last decade, which brings us to the present, has been marked by studies that benefited from the breakthroughs of the 1980s. which enabled many different laboratories to examine various aspects of apoB structure, function, and expression. The function of apoB in vivo was analyzed in different animal models (e.g., transgenic animals that overexpress apoB) (Linton et al., 1993; Callow and Rubin, 1995; Veniant et al., 1997) and in knockout animals that have no functional apoB (Farese et al., 1995,1996; Huang et al., 1995,1996). Furthermore, the structure-function relationship of apoB has been investigated in mice that express site-specific apoB mutants (Callow and Rubin, 1995; Veniant et al., 1997: Boren et al., 1998). A breakthrough in a related area led to the identification and cloning of microsomal triglyceride transfer protein (MTP) (Wetterau and Zilversmitt, 1984: Wetterau et al., 1992; Sharp et al., 1993) and the demonstration that MTP is essential for apoB production (Gordon et al., 1994; Leiper et al., 1994). The absence of MTP was found to lead to the complete degradation of apoB, which harks back to an observation in 1987 that, even in the presence of MTP, a substantial proportion of newly synthesized apoB-100 undergoes intracellular degradation before secretion (Borchardt and Davis, 1987). Indeed, the intracellular degradation of apoB-100 is the major determinant of its production rate from the liver, since the transcription of apoB appears to be constitutive and not subject to much regulation (Pullinger et al., 1989). It was in 1996, almost a decade after the first description of apoB's destruction inside the cell, that the proteasome-ubiquitin pathway was found to be the major mechanism for the intracellular degradation of apoB-100 (Yeung et al., 1996). Another important development within the last decade was the cloning of APOBEC-1, the catalytic subunit of the apoB mRNA editing complex (editosome) (Teng et al., 1993). This chapter will review some of the major landmarks in apoB research in the last 10 to 15 years, concentrating mainl PMID- 11036936 TI - Mechanism of capacitative Ca2+ entry (CCE): interaction between IP3 receptor and TRP links the internal calcium storage compartment to plasma membrane CCE channels. AB - Activation of cells by agents that stimulate inositol trisphoshate (IP3) formation causes, via IP3 receptor (IP3R) activation, the release of Ca2+ from internal stores and also the entry of Ca2+ via plasma membrane cation channels, referred to as capacitative Ca2+ entry or CCE channels. Trp proteins have been proposed to be the unitary subunits forming CCE channels; however, there is no definitive proof for this hypothesis. We have now identified amino acid sequences of a Trp and of an IP3R that interact to form stable complexes. These complexes appear to form in vivo, as evidenced by co-immunoprecipitation of Trp with IP3R and by the fact that expression of the respective interacting sequences modulates development of CCE brought about by store depletion. The finding that a Trp interacting sequence of IP3R interferes with natural CCE leads us to conclude that Trp proteins are, indeed, structural members of CCE channels. We conclude further that direct coupling of IP3R to Trp is a physiological mechanism by which cells trigger CCE in response to signals that stimulate phosphoinositide hydrolysis and IP3 formation. Pros and cons of various CCE activation models are discussed. PMID- 11036937 TI - Estrogen receptors: selective ligands, partners, and distinctive pharmacology. AB - The action of nuclear hormone receptors is tripartite, involving the receptor, its ligands, and its co-regulator proteins. The estrogen receptor (ER), a member of this superfamily, is a hormone-regulated transcription factor that mediates the effects of estrogens and anti-estrogens (e.g., tamoxifen) in breast cancer and other estrogen target cells. This chapter presents our recent work on several aspects of estrogen action and the function of the ER: 1) elucidation of ER structure-function relationships and development of ligands that are selective for one of the two ER subtypes, ERalpha or ERbeta; 2) identification of ER selective co-regulators that potentiate the inhibitory effectiveness of anti estrogens and dominant-negative ERs and modulate the activity of estrogens; 3) characterization of genes that are regulated by the anti-estrogen-ER versus the estrogen-ER complex; and 4) elucidation of the intriguing pharmacology of these ER complexes at different gene regulatory sites. These findings indicate that different residues of the ER hormone-binding domain are involved in the recognition of structurally distinct estrogens and anti-estrogens and highlight the exquisite precision of the regulation of ER activities by ligands, with small changes in ligand structure resulting in major changes in receptor character. Studies also explore the biology and distinct pharmacology mediated by ERalpha and ERbeta complexed with different ligands through different target genes. The upregulation of the anti-oxidant detoxifying phase II enzyme, quinone reductase, by the anti-estrogen-occupied ER, mediated via the electrophile response element in the QR gene, may contribute to the beneficial antioxidant effects of anti estrogens in breast cancer and illustrates the activation of some genes by ER via non-estrogen response element sequences. The intriguing biology of estrogen in its diverse target cells is thus determined by the structure of the ligand, the ER subtype involved, the nature of the hormone-responsive gene promoter, and the character and balance of co-activators and co-repressors that modulate the cellular response to the ER-ligand complex. The continuing development of novel ligands and the study of how they function as selective agonists or antagonists through ERalpha or ERbeta should allow optimized tissue selectivity of these agents for hormone replacement therapy and treatment and prevention of breast cancer. PMID- 11036938 TI - Glia-to-neuron signaling and the neuroendocrine control of female puberty. AB - It is becoming increasingly clear that astroglial cells are active participants in the process by which information is generated and disseminated within the central nervous system (CNS). In the hypothalamus, astrocytes regulate the secretory activity of neuroendocrine neurons. They contribute to facilitating sexual development by stimulating the release of luteinizing hormone-releasing hormone (LHRH), the neuropeptide that controls sexual development, from LHRH neurons. Astrocytes secrete several growth factors able to stimulate LHRH secretion. Two members of the epidermal growth factor (EGF) family--transforming growth factor alpha (TGFalpha) and the neuregulins (NRGs)-are produced in hypothalamic astrocytes and elicit LHRH secretion indirectly, via activation of receptor complexes formed by three members of the EGF receptor family, also located on astrocytes. Activation of these receptors results in the production of at least one neuroactive substance, prostaglandin E2 (PGE2), which stimulates LHRH secretion upon binding to specific receptors on LHRH neurons. Overexpression of TGFalpha in the hypothalamus accelerates puberty, whereas blockade of either TGFalpha or NRG actions delays the process, indicating that both peptides are physiological components of the neuroendocrine mechanism that controls sexual maturation. An increase in hypothalamic expression of at least two of the erbB receptors is initiated before the pubertal augmentation of gonadal steroid secretion and is completed on the day of the first preovulatory surge of gonadotropins. This secondary increase is brought about by gonadal steroids. Estrogen and progesterone facilitate erbB-mediated glia-to-LHRH neuron communication by enhancing astrocytic gene expression of at least one of the EGF related ligands (TGFalpha) and two of the receptors (erbB-2 and erbB-4). They also facilitate the LHRH response to PGE2 via induction of PGE2 receptors in LHRH neurons. A search for genes that may act as upstream regulators of the pubertal process resulted in the identification of two potential candidates: Oct-2, a POU domain gene originally described in cells of the immune system, and TTF-1, a member of the Nkx family of homeodomain transcriptional regulators required for diencephalic morphogenesis. The hypothalamic expression of both genes increases during juvenile development before the first hormonal manifestations of puberty take place. Their mRNA transcripts are localized to specific hypothalamic cellular subsets, where they appear to regulate different, but interactive, components of the neuronal-glial complex controlling LHRH secretion. While Oct-2 transactivates the TGFalpha promoter, TTF-1 does so to the erbB-2 and LHRH genes but inhibits preproenkephalin promoter activity, suggesting that both transcriptional regulators may act coordinately in the normal hypothalamus to activate genes involved in facilitating the advent of puberty and repress those restraining sexual development. Altogether, these observations indicate that the central activation of the pubertal process involves the participation of both neuronal and astroglial networks and the contribution of upstream transcriptional regulators acting on both the neuronal and glial components of the system. PMID- 11036939 TI - New steps in the Wnt/beta-catenin signal transduction pathway. AB - Wnt regulates developmental and oncogenic processes through its downstream effector, beta-catenin, and a set of other intracellular regulators that are largely conserved among species. Wnt family genes encode secreted glycoproteins that act as ligands for membrane receptors belonging to the Frizzled family of proteins. Wnt-1 originally was found as a proto-oncogene that was upregulated in tumors caused by the mouse mammary tumor virus. The Drosophila homologue of Wnt 1, wingless, is a segment polarity gene that regulates body patterning of the fly embryo. In Xenopus, the Wnt pathway regulates formation of the ventral-dorsal axis. Although Wnt proteins are expressed widely in mammals, the function of the Wnt signaling pathway in normal adult mammalian tissues is not understood. Downstream components of the Wnt pathway, APC (adenomatous polyposis coli) and beta-catenin, clearly are involved in human cancer. There are also several reports that Wnt ligands are highly expressed in tumors. Wnt stabilizes cytoplasmic beta-catenin and activates beta-catenin/Lef-1 (lymphoid enhancer factor), Tcf (T-cell factor)-dependent gene transcription. This regulation of cytosolic beta-catenin is mediated by glycogen synthase kinase-3 (GSK-3) activity but in neither case is the mechanism known. The mechanism by which Wnt inhibits GSK-3 is unknown. Recent studies have shown that some of the intracellular signaling molecules that mediate the Wnt pathway are in complexes, including Dishevelled (Dsh or Dvl), GSK-3beta, and APC protein. However, little is known about how Wnt or other upstream stimuli regulate these complexes to stabilize beta-catenin. We took a variety of approaches to identify new components of the Wnt pathway. Using an expression-cloning technique, we isolated casein kinase I (CKI)epsilon as a positive regulator of beta-catenin in the Wnt pathway. Overexpression of CKIepsilon mimics Wnt by stabilizing beta-catenin, thereby increasing expression of beta-catenin-dependent genes. Inhibition of endogenous CKIepsilon attenuated gene transcription stimulated by Wnt or by Dsh. CKIepsilon forms a complex with Axin and the other downstream components of the Wnt pathway. CKIepsilon is a positive regulator of the Wnt pathway and a possible functional link between upstream signals and the intracellular Axin signaling complex that regulates beta-catenin. In separate experiments, we have identified a Dishevelled associated kinase (DAK) that binds to Dsh and regulates its functions. Dsh is required for two different pathways, the Wnt pathway and planar polarity pathway in Drosophila. DAK dramatically enhances the function of Dsh in the Wnt pathway and inhibits its function in the planar polarity pathway. This chapter will discuss these newly identified components of the Wnt pathway. PMID- 11036940 TI - Regulation of the pituitary somatotroph cell by GHRH and its receptor. AB - Hormones from the hypothalamus mediate interactions between the nervous and endocrine systems by controlling the activity of specific target cells in the anterior pituitary gland. The hypothalamic peptide, growth hormone-releasing hormone (GHRH), acts on pituitary somatotroph cells to stimulate their proliferation during development and to regulate their ability to produce and secrete growth hormone (GH). These actions are mediated by a recently identified receptor for GHRH that belongs to family B-III of the G protein-coupled receptor superfamily. The rat GHRH receptor is expressed predominantly in the pituitary gland and in somatotroph cells. To investigate this tissue- and cell-specific expression, the receptor gene has been cloned and characterized. The receptor gene promoter is selectively expressed in pituitary cells and is regulated by the pituitary-specific transcription factor Pit-1. There is a sexual dimorphism in GHRH receptor expression in the rat pituitary, suggesting regulation by gonadal steroids. In addition, glucocorticoids are potent positive regulators of GHRH receptor gene expression. Substantial evidence points to an important role for GHRH in regulating the proliferation and functional activity of the somatotroph cell. This is best observed in the dwarf little mouse, which harbors a mutation in the extracellular domain of the GHRH receptor that abolishes the receptor's hormone-binding and signaling properties, resulting in severe somatotroph hypoplasia. Complementary studies in transgenic mice overexpressing the ligand GHRH reveal corresponding somatotroph hyperplasia. Consistent with these observations, GHRH potently activates the MAP kinase pathway in pituitary somatotroph cells. To better understand the hormone-binding and signaling properties of the GHRH receptor, mutant and chimeric receptors have been analyzed to define domains important for GHRH interaction. The GHRH receptor signals predominantly through cAMP-dependent pathways; however, a variant form of the GHRH receptor with an insertion into the third intracellular domain, generated through alternative RNA processing, binds GHRH but fails to signal, suggesting potential modulation of receptor function at a post-transcriptional level. This chapter will integrate these basic investigations of GHRH and its receptor with current information on the involvement of the GHRH signaling system in human diseases of GH secretion and growth. PMID- 11036941 TI - Secretagogues and the somatotrope: signaling and proliferation. AB - Somatotrope function requires consideration of both growth hormone (GH) secretion and cellular proliferation. The regulation of these processes is, to a large extent, controlled by three hypothalamic hormones: GH-releasing hormone (GHRH), somatostatin (SRIF), and an as-yet-unidentified GH secretagogue (GHS). Each binds to G protein-linked membrane receptors through which signaling occurs. Our laboratory has used a series of genetic and transgenic models with perturbations of individual components of the GH regulatory system to study both somatotrope signaling and proliferation. Impaired GHRH signaling is present in the lit mouse, which has a GHRH receptor (R) mutation, and the dw rat, which has a post-receptor signaling defect. Both models also have impaired responses to a GHS, implying an interaction between the two signaling systems. The spontaneous dwarf rat (SDR), in which a mutation of the GH gene results in total absence of the hormone, shows characteristic changes in the hypothalamic regulatory hormones due to an absence of GH feedback and alterations in the expression of each of their pituitary receptors. Treatment of SDRs with GHRH and a GHS has allowed demonstration of a stimulatory effect of GHRH on GHRH-R, GHS-R, and SRIF type 2 receptor (SSTR-2) expression and an inhibitory effect on SSTR-5 expression. GH also modifies the expression of these receptors, though its effects are seen at later time periods and appear to be indirect. Overall, the results indicate a complex regulation of GH secretion in which somatotrope receptor, as well as ligand expression, exerts an important physiological role. Both the SDR and the GH-R knockout (ko) mouse have small pituitaries and decreased somatotropes, despite elevated GHRH secretion and intact GHRH-R signaling. Introduction of the hGHRH transgene into GH-R ko mice confirmed that the proliferative effects of GHRH require GH/insulin like growth factor-I (IGF-I) action. The results offer new insights into factors participating in somatotrope proliferation. PMID- 11036942 TI - SH2-B and SIRP: JAK2 binding proteins that modulate the actions of growth hormone. AB - Growth hormone (GH) has long been known to be a primary determinant of body height and an important regulator of body metabolism, yet the cellular and molecular bases for these effects of GH are only beginning to be understood. In 1993, GH receptor (GHR) was first observed to bind to the tyrosine kinase JAK2. GH increased JAK2's affinity for GHR, potently activated JAK2, and stimulated the phosphorylation of tyrosines within JAK2 and the cytoplasmic domain of GHR. In the intervening six years, a variety of signaling molecules have been identified that are tyrosyl phosphorylated in response to GH, presumably by the activated JAK2. These signaling molecules include 1) the latent cytoplasmic transcription factors--designated signal transducers and activators of transcription (Stats)- that have been implicated in the regulation of a variety of GH-dependent genes; 2) Shc proteins that lead to activation of the Ras-MAP kinase pathway: and 3) insulin receptor substrate (IRS) proteins that bind and thereby activate phosphatidylinositol 3' kinase and presumably other proteins. Recently, we have identified two additional signaling molecules for GH that bind to JAK2 and are phosphorylated on tyrosines in response to GH: SH2-B and signal regulated protein (SIRP). Based upon amino acid sequence analysis, SH2-B is presumed to be a cytoplasmic adapter protein. It binds with high affinity via its SH2 domain to phosphorylated tyrosines within JAK2. GH-induced binding of SH2-B to JAK2 via this site potently activates JAK2, leading to enhanced tyrosyl phosphorylation of Stat proteins and other cellular proteins. Because of its other potential protein protein interaction domains and its recruitment and phosphorylation by kinases that are not activated by SH2-B, SH2-B is thought likely to mediate other, more specific actions of GH, as yet to be determined. SIRP is a transmembrane protein that is now known to bind to integrin-associated protein. It appears to bind directly to JAK2 by a process that does not require tyrosyl phosphorylation, although is itself highly phosphorylated on tyrosines in response to GH. The phosphorylated SIRP recruits one or more molecules of the tyrosine phosphatase SHP2 that, in turn, de-phosphorylates SIRP and most likely JAK2. Thus, SIRP is predicted to be a negative regulator of GH action. It seems likely that the diverse actions of GH will be found to require coordinated interaction of all of these signaling proteins with each other as well as with other signaling molecules that are activated by GH and the numerous other ligands that are present at cells during a response to GH. PMID- 11036943 TI - VIII International Workshop on Chronic Lymphocytic Leukaemia. 29-31 October 1999, Paris, France. Proceedings and abstracts. PMID- 11036944 TI - Genetics of chronic lymphocytic leukaemia. PMID- 11036945 TI - CLL variants and T-cell-related issues. PMID- 11036946 TI - Cellular interactions, immunodeficiency and autoimmunity in CLL. PMID- 11036947 TI - Chronic lymphocytic leukemia. Preclinical oral poster session. PMID- 11036948 TI - Current therapy for CLL. PMID- 11036949 TI - Chronic lymphocytic leukemia . Clinical oral poster session. PMID- 11036950 TI - Emerging therapies and future directions in chronic lymphocytic leukaemia- chemotherapy. PMID- 11036951 TI - Emerging therapies and future directions in CLL--monoclonal antibodies, vaccines and gene therapy. PMID- 11036953 TI - Telomerase inhibitors: targeting the vulnerable end of cancer? AB - In the past decade, a great deal has been learnt about the maintenance of telomeres in mammalian cells by the specialized reverse transcriptase, telomerase, and its associated proteins. The catalytic component of telomerase, hTERT, appears to be selectively activated in the vast majority of tumors relative to most somatic cells suggesting that its inhibition may result in antitumor effects. Although beset with some unusual issues as a drug target, recent 'target validation' studies using hTERT dominant-negative and antisense approaches strongly support the view that potent and selective telomerase inhibitors will induce inhibitory effects on tumors, especially in those possessing relatively short telomeres. Inhibitory strategies have focused on three main areas: antisense molecules (oligonucleotides, RNA molecules, ribozymes and peptide nucleic acids) directed against the hTR RNA component of telomerase, small molecule reverse transcriptase inhibitors (e.g. azidothymidine), and, probably most advanced, small molecules capable of interacting with and stabilizing four-stranded (G-quadruplex) structures formed by telomeres. G quadruplex interactive agents that inhibit telomerase at sub-micromolar concentrations in cell-free assays have been described. Lead optimization and preclinical whole-cell and animal antitumor and pharmacology studies are now progressing which should result in the first generation of telomerase inhibitors being evaluated in the clinic within the next few years. PMID- 11036954 TI - The clinical pharmacology of alkylating agents in high-dose chemotherapy. AB - Alkylating agents are widely used in high-dose chemotherapy regimens in combination with hematological support. Knowledge about the pharmacokinetics and pharmacodynamics of these agents administered in high doses is critical for the safe and efficient use of these regimens. The aim of this review is to summarize the clinical pharmacology of the alkylating agents (including the platinum compounds) in high-dose chemotherapy. Differences between conventional and high doses will be discussed. PMID- 11036955 TI - Phase II study of bendamustine in patients with relapsed or cisplatin-refractory germ cell cancer. AB - Despite generally high cure rates in patients with metastatic germ cell cancer, patients with incomplete response to first-line cisplatin-based chemotherapy or with relapsed disease following high-dose salvage therapy exhibit a very poor prognosis. We investigated the efficacy and toxicity of bendamustine, a bifunctional alkylating benzimidol derivative with only partial cross-resistance to other alkylating agents such as ifosfamide or cyclophosphamide. Nineteen patients with cisplatin-refractory germ cell tumors (GCT) or relapse after high dose chemotherapy plus autologous stem cell support were treated with bendamustine at a dose of 120 mg/m2 on 2 consecutive days at 3 week intervals. Patients had received a median of 9 (range 4-20) platinum-containing treatment cycles prior to bendamustine and 13 patients (68%) had previously received carboplatin/etoposide-based high-dose chemotherapy. One patient achieved a partial remission of only 6 weeks duration. No other responses were seen. Toxicity was low with one patient developing WHO grade 3 thrombocytopenia as the only WHO grade 3/4 toxicity observed. Hematologic toxicity was similar in patients pretreated with and without high-dose chemotherapy plus autologous stem cell support. We conclude that bendamustine has little or no clinically relevant activity in patients with cisplatin-refractory GCT or relapsed disease after high dose chemotherapy. PMID- 11036956 TI - Dose-finding study of paclitaxel and carboplatin in patients with advanced non small cell lung cancer. AB - This dose-finding study was designed to determine the maximum tolerated dose (MTD), efficacy and toxicity of combined paclitaxel and carboplatin in 35 previously untreated patients with advanced non-small cell lung cancer (NSCLC). Paclitaxel was given as a 3-h infusion at escalating dose levels (100-250 mg/m2) immediately followed by carboplatin as a 30-min infusion (325 or 350 mg/m2) every 3 weeks. The dose-limiting toxicity, paresthesia, occurred at the highest dose level, therefore the recommended dose was established one level below (paclitaxel 225 mg/m2 with carboplatin 325 mg/m2). Neutropenia was the most common hematotoxicity; dose dependency was not apparent. Two patients, at different dose levels, had febrile neutropenia. Thrombocytopenia was rare. Non-hematological toxicities grade 3 or higher included infection, anorexia, alopecia and paresthesia. One patient had a hypersensitivity reaction (transient hypotension). The overall response rate was 23% and median survival time was 7.5 months. Promising activity and acceptable toxicity supports the development of this combination as a useful chemotherapeutic option in advanced NSCLC. PMID- 11036957 TI - Adjuvant intraportal chemotherapy for Dukes B2 and C colorectal cancer also receiving systemic treatment: results of a multicenter randomized trial. Groupe Regional d'Etude du Cancer Colo-Rectal (Belgium). AB - In a randomized trial, the authors evaluated the possible adjuvant activity of intraportal chemotherapy (with 5-fluorouracil 500 mg/m2/day in continuous infusion for 7 days and mitomycin C 10 mg/m2 at day 7) administered after surgery to half of the patients who underwent a full resection for Dukes B2 or C colorectal cancer. The procedure appeared manageable and safe. Two hundred and sixty patients were initially randomized, among whom 173 were finally considered as fully evaluable after having completed six courses of systemic chemotherapy. The reasons for withdrawal were basically tumoral ones and patients or doctors compliance. After a median follow-up of 4.5 years, no difference could be observed in the patients evolution assessed as relapses or deaths rate, or as relapse-free (at 5 years: 68% in the portal treatment group versus 70% in the control group) or overall survival (at 5 years: 76 versus 74%). The frequency of hepatic metastases (21 versus 18%) was also similar in both groups. PMID- 11036958 TI - Inhalational interleukin-2 liposomes for pulmonary metastases: a phase I clinical trial. AB - The lung is a common site of both metastases and primary neoplasia. This phase I study was designed to test the feasibility and toxicity of administering interleukin (IL)-2 liposomes by aerosol to patients with pulmonary metastases. The goal was to test whether IL-2 liposomes could be given by aerosol using biologically effective but non-toxic doses in an outpatient setting. Liposomes containing IL-2 or placebo (buffer) were synthesized and mixed to provide a constant lipid dose, and were nebulized using a Puritan twin jet nebulizer and a standard compressor. The liposome-containing mist was inhaled for about 20 min 3 times a day in order to selectively stimulate immune function within the lung and to avoid systemic toxicity. The dose chosen was based on canine efficacy and toxicity studies that used bronchoalveolar lavage to demonstrate increased cell numbers and activation of mononuclear cells after inhalation of nebulized IL-2 liposomes. Nine patients were treated in three cohorts of three patients at 1.5, 3.0 and 6.0 x 10(6) IU of IL-2 3 times a day. No significant toxicity was observed. We conclude that the delivery of IL-2 liposomes by inhalation is well tolerated. Further studies of inhalational IL-2 liposomes to determine efficacy as an anti-cancer therapy are warranted. PMID- 11036959 TI - Use of docetaxel (Taxotere) in patients with paclitaxel (Taxol) hypersensitivity. AB - Anaphylaxis or significant hypersensitivity reaction is one of the most catastrophic potential complications of chemotherapy. There is a 2-5% risk of hypersensitivity with paclitaxel, a commonly used chemotherapeutic agent for various cancers. Three patients, who developed hypersensitivity to paclitaxel infusion, received docetaxel without allergic reactions. Docetaxel may therefore be an alternative treatment for patients with paclitaxel hypersensitivity. PMID- 11036960 TI - Paclitaxel-induced remission in docetaxel-refractory anthracycline-pretreated metastatic breast cancer. AB - Paclitaxel and docetaxel are excellent agents with a high antitumor effect for the treatment of previously anthracycline-exposed metastatic breast cancer. There has been no standard treatment for patients who undergo therapy of a taxan resistant metastatic breast cancer. Paclitaxel has partial non-cross-resistance in vitro with docetaxel in inhibiting microtubule disaggregation. We present the case of a patient with docetaxel-refractory anthracycline-pretreated metastatic breast cancer who achieved remission with paclitaxel. PMID- 11036961 TI - In vitro sensitivity of human endometrial cancer cell lines to paclitaxel or irinotecan (CPT-11) in combination with other aniticancer drugs. AB - We have evaluated the growth inhibitory effects of paclitaxel alone or irinotecan (CPT-11) alone and their combined effects with other drugs on human endometrial cancer cell lines. IC50 doses of paclitaxel (Tx), SN-38 (active metabolite of CPT 11; 7-ethyl-10-hydroxycamptothecin) and cisplatin, including other drugs which have been used for treatment of patients with endometrial cancer, were examined using five human endometrial cancer cell lines (Ishikawa, HEC-1A, HEC-50B, HEC-59 and HEC-108). When in vitro sensitivity was defined IC50 lower than 10% of the peak plasma concentration (PPC), all endometrial cancer cell lines were sensitive to paclitaxel and three of five endometrial cancer cell lines were sensitive to SN-38, whereas cisplatin was not active against any endometrial cancer cell lines used in this study. Regarding the other drugs, aclarubicin (ACR) and actinomycin D (ACD) were active against four of five endometrial cancer cell lines, etoposide (VP-16) and pirarubicin (THP) against two, and 5-fluorouracil (5-FU) against only one, while ifosfamide (4-OHIFO) was not active against any endometrial cancer cell lines. When combined effects of paclitaxel or SN-38 with other one drug were determined by the median-effect analysis, paclitaxel followed by cisplatin resulted in synergistic effects to all endometrial cancer cell lines. Paclitaxel followed by SN-38 also had synergistic effects to four cell lines. Sequential but not simultaneous administration of taxol and THP-adriamycin showed synergistic effects to three cell lines. In combinations of SN-38 with other drugs, simultaneous administration of SN-38 and cisplatin resulted in synergistic effects to all cell lines. It is noteworthy that ACD followed by SN-38 showed synergistic effects to all cell lines, and simultaneous treatment of ACD and SN 38 or SN-38 followed by ACD also resulted in synergistic effects to three cell lines. THP-adriamycin followed by SN-38 had synergistic effects to four cell lines. The present quantitative data analysis for synergism provides a basis for a rational design of clinical protocols for combination chemotherapy in patients with endometrial cancer of the uterus. PMID- 11036962 TI - Synergistic antitumoral activity of combined UFT, folinic acid and oxaliplatin against human colorectal HT29 cell xenografts in athymic nude mice. AB - This study was designed to assess the inhibition of tumor growth by oxaliplatin combined with UFT and folinic acid (FA). Growth inhibition was studied in nude mice transplanted with human colorectal HT29 tumor cell xenografts and treated for 28 days with oral UFT (20 mg/kg/day) and FA (4 mg/kg/day), i.p. oxaliplatin (10 mg/kg on day 1) or a combination of oxaliplatin, UFT and FA, or else not treated (controls). Tumor surface area and weight were recorded twice a week, and mice were sacrificed at day 28. Two separate experiments were performed for each group of 25 mice. At day 28, mean tumor weights (g) were 2.89+/-0.22 (controls), 2.03+/-0.14 (oxaliplatin), 2.02+/-0.21 (UFT/FA) and 1.23+/-0.17 (oxaliplatin+UFT/FA). For the three treatment groups, tumor weight decreases were 30.1% (p<0.05), 29.9% (p<0.05) and 57.5% (p<0.001), respectively. Combined treatment (UFT/FA+oxaliplatin) reduced tumor weight by 39% compared to oxaliplatin alone (p<0.05) or UFT/FA (p<0.05). These results demonstrate the synergistic effect of the combination of oxaliplatin, UFT and FA in this HT29 cell xenograft model, and warrant further investigations in patients with metastatic colorectal cancer. PMID- 11036963 TI - A bioassay for the activity of PSC 833 in human serum for modulation of P glycoprotein-mediated multidrug resistance. AB - We established a rapid and sensitive ex vivo bioassay to detect the multidrug resistance (MDR)-inhibitory activity of SDZ PSC 833 ([3'-keto-Bmt1]-[Val2] cyclosporin (PSC 833)) in two RPMI 8226 human myeloma sublines (parent 8226 and doxorubicin-resistant subline Dox6) in 75% human serum. In vitro sensitivity of the tumor to doxorubicin was determined by 3-h drug exposure growth inhibition assay (MTT assay). PSC 833 in serum restored the IC50 of doxorubicin in the P glycoprotein (P-gp)-positive resistant subline to the same level as in the sensitive cells at 1 microg/ml, which has been shown to be an achievable concentration in clinical trials. In addition, the cytotoxic effect of doxorubicin was enhanced by PSC 833 in the sera of the patient in whom the blood level was 705.7 ng/ml. However, 10 microg/ml PSC 833 in serum does not cause a complete recovery in the IC90 of doxorubicin in the resistant sublines. This MDR inhibitory activity was supported by the finding that PSC 833 in serum does not increase accumulation of rhodamine 123 in doxorubicin-resistant cells in an in vitro functional assay. The present study provides evidence that PSC 833 in human serum is effective to modulate P-gp-mediated MDR but insufficient for the reversal of MDR from the clinicopharmacological point of view. PMID- 11036964 TI - Cost-utility analysis of second-line hormonal therapy in advanced breast cancer: a comparison of two aromatase inhibitors to megestrol acetate. AB - Randomized trials comparing the aromatase inhibitors, anastrozole and letrozole, to megestrol acetate (MA) in postmenopausal women with advanced breast cancer demonstrated that both agents are better tolerated than MA with comparable efficacy. In addition, one trial revealed that tumor response and time to treatment failure were significantly better with letrozole. Since oncologists are faced with a choice between three agents with at least comparable efficacy but different toxicity profiles and cost, a cost-utility analysis was conducted to quantify these differences and to determine if the new agents are more cost effective than MA. In the absence of a randomized three-arm trial, a decision model was developed to simulate the most common therapeutic outcomes. The clinical data were obtained from an overview analysis of randomized trials. Total hospital resource consumption was collected from 87 patients with advanced disease that had failed second-line hormonal therapy. Utility estimates were obtained from interviewing a random sample of 25 women from the general public and 25 female health care professionals using the Time Trade-Off technique. The model suggested a similar duration of quality-adjusted progression-free survival between drugs (letrozole 150 days, anastrozole 153 days and MA 146 days). Letrozole had an overall cost of Can$2949 per patient which was comparable to MA at Can$2966 per patient. In contrast, anastrozole was slightly more costly than MA at $Can3149 per patient, respectively. The analysis revealed that letrozole has comparable overall costs relative to MA while providing at least equivalent quality-adjusted progression-free survival. These outcomes were largely related to its higher tumor response rate, which translated to a lower proportion of patients requiring chemotherapy. Anastrozole was slightly more costly than MA and did not demonstrate superiority in quality-adjusted progression-free survival in this palliative setting. PMID- 11036965 TI - Analysis of protein self-association under conditions of the thermodynamic non ideality. AB - Analysis of protein-protein interactions in highly concentrated solutions requires a consideration of the non-ideality in such solutions which is expressed by the virial coefficients. Different equations are presented to estimate effects of the thermodynamic non-ideality on the macromolecular interaction of self associating proteins in sedimentation equilibrium experiments. Usually the influence of thermodynamic non-ideal behavior are described by concentration power series. The convergence of such power series is limited at high solute concentration. When expressing the thermodynamic non-ideality by an activity power series this disadvantage can be minimized. The developed centrifuge equations are the basis for a global analysis to estimate equilibrium constants and the corresponding thermodynamic activities of the reactants. Based on fit analysis of synthetic concentration profiles it was established that marked deviations from the expected association constants are observed for proteins with strong association forces between solute molecules. Considerable differences were also observed in weakly interacting systems. This was due to the excluded volume of the protein which is similar in magnitude to the binding constant. For interactions with moderate affinities values extremely close to the true binding values were obtained, as confirmed by experimental results with concanavalin A. PMID- 11036966 TI - Oxygen and acrylamide quenching of protein phosphorescence: correlation with protein dynamics. AB - Oxygen quenching of protein phosphorescence and activation enthalpies for the structural fluctuations underlying O2 and acrylamide diffusion were determined for RNase T1, glyceraldehyde-3-phosphate dehydrogenase and beta-lactoglobulin, which have the phosphorescing residues located in relatively solvent-exposed and flexible regions of the polypeptide. The results, compared with those obtained for proteins characterised by a very rigid environment, established that kqO2 was directly correlated to the flexibility of the protein matrix surrounding the chromophore. While the migration of acrylamide was characterised by delta H(double dagger), which was strongly dependent on the fluidity of the structure about the Trp residue, the values of the activation enthalpies for the oxygen migration of all the proteins studied were rather similar, approximately 10 kcal mol(-1), in spite of the depth of the chromophore and the rigidity of its environment. The implications of these findings for the migration of small solutes inside proteins have been discussed. PMID- 11036967 TI - Effect of local anesthetics on the bilayer membrane of dipalmitoylphosphatidylcholine: interdigitation of lipid bilayer and vesicle micelle transition. AB - The phase transitions of dipalmitoylphosphatidylcholine (DPPC) bilayer membrane were observed by means of differential scanning calorimetry (DSC) as a function of the concentration of local anesthetics, dibucaine (DC x HCl), tetracaine (TC x HCl), lidocaine (LC x HCl) and procaine hydrochlorides (PC x HCl). LC x HCl and PC x HCl depressed monotonously the temperatures of the main- and pre-transition of DPPC bilayer membrane. The enthalpy changes of both transitions decreased slightly with an increase in anesthetic concentration up to 160 mmol kg(-1). In contrast, the addition of TC x HCl or DC x HCl, having the ability to form a micelle by itself, induced the complex phase behavior of DPPC bilayer membrane including the vesicle-to-micelle transition. The depression of both temperatures of the main- and pre-transition, which is accompanied with a decrease in enthalpy, was observed by the addition of TC x HCl up to 21 mmol kg(-1) or DC x HCl up to 11 mmol kg(-1). The pretransition disappeared when these concentrations of anesthetic were added, and the interdigitated gel phase appeared above these concentrations. The appearance of the interdigitated gel phase, instead of the ripple gel phase, brings about the stabilization of the gel phase by 1.8-2.4 kcal mol(-1). In the concentration range of 70-120 mmol kg(-1) TC x HCl (or 40-60 mmol kg(-1) DC x HCl), the enthalpy of the main transition exhibited a drastic decrease, resulting in the virtual disappearance of the main transition. This process includes the decrease in vesicle size with increasing anesthetic concentration, resulting in the mixed micelle of DPPC and anesthetics. Therefore, in this range of anesthetic concentration, the DPPC vesicle solubilized an anesthetic which coexists with the DPPC-anesthetic mixed micelle. Above the concentration of 120 mmol kg(-1) TC x HCl (or 60 mmol kg(-1) DC x HCl), there exists the DPPC-anesthetic mixed micelle. Two types of new transitions concerned with the mixed micelle of DPPC and micelle-forming anesthetics were observed by DSC. PMID- 11036968 TI - Depth-dependent analysis of membranes using benzophenone-based phospholipids. AB - Any attempt to probe the membrane hydrophobic core with chemical reagents necessitates the use of reactive intermediates like carbenes and nitrenes, which can insert into C-H bonds. Several photoactivable reagents based on carbenes and nitrenes have been reported. However, the high reactivity of these reagents, often leads to very low insertion yields. We report here a high degree of cross linking (35-40%) achieved with three benzophenone-based phospholipids and analyze the carbon functionalization data using a multiple Gaussian function. These phospholipids are so designed so as to permit depth-dependent labeling in membranes. Single bilayer vesicles were prepared from these phospholipids and dimyristoylphosphatidylcholine. The cross-linked product was isolated and characterized by mass spectroscopy. The results obtained indicated that the cross linked product was dominated by dimeric product formed by intermolecular cross linking. The Gaussian analysis used here provides insight into the relative depths of the probes inside the membrane. PMID- 11036969 TI - Microscopic model of protein crystal growth. AB - A microscopic, reversible model to study protein crystal nucleation and growth is presented. The probability of monomer attachment to the growing crystal was assumed to be proportional to the protein volume fraction and the orientational factor representing the anisotropy of protein molecules. The rate of detachment depended on the free energy of association of the given monomer in the lattice, as calculated from the buried surface area. The proposed algorithm allowed the simulation of the process of crystal growth from free monomers to complexes having 10(5) molecules, i.e. microcrystals with already formed faces. These simulations correctly reproduced the crystal morphology of the chosen model system--the tetragonal lysozyme crystal. We predicted the critical size, after which the growth rate rapidly increased to approximately 50 protein monomers. The major factors determining the protein crystallisation kinetics were the geometry of the protein molecules and the resulting number of kinetics traps on the growth pathway. PMID- 11036970 TI - Structure of a fusion peptide analogue at the air-water interface, determined from surface activity, infrared spectroscopy and scanning force microscopy. AB - We have investigated a point mutant of the HIV-1 fusion peptide in a compressed monolayer at the air-water interface. A variety of surface sensitive techniques were applied to study structural features under conditions mimicking the hydrophobic/hydrophilic environment of a biomembrane. Possible partitioning into the aqueous bulk phase and molecular areas were examined by surface activity based mass conservation plots. This shows that the peptide is practically fully accumulated in the interface. Secondary structure and orientation was analyzed by means of polarized infrared reflectivity. Brewster angle microscopy and scanning force microscopy contributed nanostructural images. At low surface pressures the molecules form anti-parallel beta-sheets lying flat on the interface. Upon a moderate increase of the lateral pressure a flat beta-turn structure appears with inter- and intramolecular H-bonds. We also observed aggregates forming fingerprint-like structures with a diameter of approximately double the hydrophobic length of a beta-turn conformation. Beyond approximately 18 mN m(-1) the beta-turns straighten up. The lowest measured tilt angle was 45 degrees at 36 mN m(-1). PMID- 11036971 TI - Fluorescence dynamics of green fluorescent protein in AOT reversed micelles. AB - We have used the enhanced green fluorescent protein (EGFP) to investigate the properties of surfactant-entrapped water pools in organic solvents (reversed micelles) with steady-state and time-resolved fluorescence methods. The surfactant used was sodium bis(2-ethylhexyl)sulfosuccinate (AOT) and the organic solvents were isooctane and (the more viscous) dodecane, respectively. The water content of the water pools could be controlled through the parameter w0, which is the water-to-surfactant molar ratio. With steady-state fluorescence, it was observed that subtle fluorescence changes could be noted in reversed micelles of different water contents. EGFP can be used as a pH-indicator of the water droplets in reversed micelles. Time-resolved fluorescence methods also revealed subtle changes in fluorescence decay times when the results in bulk water were compared with those in reversed micelles. The average fluorescence lifetimes of EGFP scaled with the relative fluorescence intensities. Time-resolved fluorescence anisotropy of EGFP in aqueous solution and reversed micelles yielded single rotational correlation times. Geometrical considerations could assign the observed correlation times to dehydrated protein at low w0 and internal EGFP rotation within the droplet at the highest w0. PMID- 11036972 TI - Evaluation of total mercury concentrations in fish consumed in the municipality of Itaituba, Tapajos River Basin, Para, Brazil. AB - The environmental exposure to mercury by riverine and Indian communities can occur through the ingestion of fish and products contaminated by mercury compounds. The present study aims to evaluate the total mercury concentration in the different fish species most consumed in the municipality of Itaituba, Tapajos river basin, where there are intense gold-mining activities. These fish samples were analyzed by atomic absorption spectrophotometry by cold vapor technique, Automatic Mercury Analyzer HG-3500. The largest mercury concentrations were found in the carnivorous species ranging from 112.4 to 2250 microg/g, while the detritivorous, herbivorous and omnivorous species presented total mercury levels ranging from 3.2 to 309.8 microg/g, which is below the limit established by the World Health Organization. This paper also reports and identifies which species are more polluted and present a statistical relationship among concentration and weight of the carnivorous species studied in detail, Brachyplatystoma flavicans (dourada) (r2 = 0.691) and Pseudoplatystoma sp. (surubim) (r2 = 0.654). PMID- 11036973 TI - Fish mercury concentration in the Alto Pantanal, Brazil: influence of season and water parameters. AB - The tropical flood plain Pantanal is one of the world's largest wetlands and a wildlife sanctuary. Mercury (Hg) emissions from some upstream gold mining areas and recent findings of high natural Hg levels in tropical oxisols motivated studies on the Hg cycle in the Pantanal. A survey was made on total Hg in the most consumed piscivorous fish species from rivers and floodplain lakes in the north (Caceres and Barao de Melgaco) and in the south part of Alto Pantanal (around the confluence of the Cuiaba and Paraguai rivers). Samples were collected in both the rainy and dry seasons (March and August 1998) and included piranha (Serrasalmus spp.), and catfish (Pseudoplatystoma coruscans, pintado, and Pseudoplatystoma fasciatum, cachara or surubim). There was only a small spatial variation in Hg concentration of the 185 analyzed fish samples from the 200 x 200 km large investigation area, and 90% contained total Hg concentration below the safety limit for regular fish consumption (500 ng g(-1)). Concentration above this limit was found in both Pseudoplatystoma and Serrasalmus samples from the Baia Sia Mariana, the only acid soft-water lake included in this study, during both the rainy and dry seasons. Concentration above this limit was also found in fish outside Baia Sia Mariana during the dry season, especially in Rio Cuiaba in the region of Barao de Melgaco. The seasonal effect may be connected with decreasing water volumes and changing habitat during the dry season. The results indicate that fertile women should restrict their consumption of piscivorous fishes from the Rio Cuiaba basin during the dry season. Measures should be implanted to avoid a further deterioration of fish Hg levels. PMID- 11036974 TI - Mercury species in the liver of ringed seals. AB - Four types of mercury species, namely, methylmercury, organic mercury other than methylmercury, inorganic mercury, and insoluble mercury, deemed to be mercuric selenide (HgSe), were found in the liver of 45 ringed seals from the Canadian Arctic. On average, methylmercury, at 2%, made up the smallest fraction of the total mercury in the liver of these animals. Of the total mercury concentration in seal liver, 53% was insoluble mercury, estimated to be largely mercuric selenide. Other workers have found this compound to be present in mammalian liver and identified it to be HgSe. Organic mercury other than methylmercury made up 4%, and inorganic mercury 42% of the total mercury in the liver. The sum of the independently determined mercury species agreed well with the total mercury concentration in the liver. Species other than mercuric selenide are known to be toxic. Mercuric selenide, considered to be a stable end product of the demethylation process of methylmercury, although not readily eliminated from the liver, is inert and apparently non-toxic. Only approximately half of the total mercury in the liver was potentially toxic mercury. All four mercury species were positively correlated with the age of animals, the regression slope on age being 20 times larger for insoluble Hg (HgSe) than for methylmercury. A number of reported observations, such as the long half-life of Hg in liver (> or = 10 years), the dependence of Hg on age, and the often-observed one-to-one relationship between Hg and Se (on a molar basis), are readily explained by the temporal accumulation of HgSe in the liver. In the future, a more accurate assessment of the health risk to animals and humans from the consumption of contaminated animal tissues will be possible, by measuring all mercury species rather than just total mercury or methylmercury. Total mercury alone in the liver is an inadequate indicator of toxicity to animals. Methylmercury was analyzed by capillary gas-liquid chromatography with ECD detection, and the other species were operationally/experimentally defined using physical/chemical methods. PMID- 11036975 TI - The use of stable carbon isotopes to evaluate the importance of fine suspended particulate matter in the transfer of methylmercury to biota in boreal flooded environments. AB - Applying the classic geochemical technique of stable carbon isotope ratios (delta13C), we confirmed the existence of a trophic link between fine particulate matter (FPM) and zooplankton in freshwater ecosystems, and examined possible reasons for the elevated MeHg concentrations ([MeHg]) in hydroelectric reservoirs as compared to natural lakes. Comparing natural and flooded environments, the delta13C and [MeHg] values for FPM and zooplankton differ significantly. Using a mixing model to calculate the contribution of terrestrial carbon to FPM, the differences in delta13C between natural and flooded sites are explained by an increasingly important autochthonous component in reservoirs. The stable isotopic evidence presented here strongly suggests that, despite the much greater abundance of detrital vascular-plant carbon, microalgae are important in supporting aquatic food webs in the oligotrophic flooded systems studied. Due to a significant inverse relationship between [MeHg] in FPM and the percentage of terrestrial carbon (r2 = 0.87), we propose that the higher [MeHg] in the zooplankton of flooded sites as compared to lakes are the result of proportionally higher levels of autochthonous material (algae/bacteria; i.e. potential sources/methylators of Hg) in the FPM of reservoirs. PMID- 11036976 TI - Methylmercury in water, seston, and epiphyton of an Amazonian river and its floodplain, Tapajos River, Brazil. AB - Levels and accumulation of MeHg were characterized in filtered water, suspended organo-mineral matter, phytoplankton, zooplankton and epiphyton during the dry season and the wet season. In open water of the lentic and lotic ecosystems, the MeHg in filtered water (< 0.2 microm) was near or below the detection limit (< 0.02-0.03 ng/l). These concentrations represent < 5% of the Hgtot. content in filtered water. Inundated forests (Igapo) and macrophyte floating mats were the only sites where MeHg was significantly detected (0.07-0.24 ng/l), representing 3 22% of the Hgtot. in filtered water. MeHg concentrations in organo-mineral suspended matter (2-26 ng/g dry wt. representing 0.6-7.3% of Hgtot.) were correlated with the N content but not with the C content. Data suggest that MeHg enrichment of suspended matter is strongly influenced by the presence of degraded planktonic remains relatively rich in N and MeHg. In zooplankton, MeHg concentrations (20-140 ng/g dry wt.) increased from the dry season to the end of the wet season. This increase was followed by higher proportions of MeHg during the wet season in comparison to the dry season (15-40 to 50-70% of the Hgtot.). The epiphytic material collected from the roots of macrophyte floating mats contained 2-8.5 ng/g dry wt. of MeHg. The proportion of MeHg to Hgtot. in epiphyton (1.5-8.3%) correlated with its C and N contents. The data suggest a greater bioavailability of MeHg in the Tapaj6s River ecosystems due to the seasonal increase in water level and the consequent inundation of the floodplain. Inundation favours the development of large macrophyte floating mats which increases the bioavailability of epiphyton to herbivorous/detritivorous fish. The root zone of floating macrophytes and the flooded organic horizon of the Igapo forest are the only sites along the Tapajos River where significant MeHg can be detected in the water column and sediment. This new study supports the hypothesis that MeHg production and transfer to the first link of the food chain in Amazonian river systems is closely related to organic matter biogeochemistry in the floodplain environment. PMID- 11036977 TI - Biomonitoring of mercury in the Kastela Bay using transplanted mussels. AB - Kastela Bay, located on the central part of east Adriatic coast, is heavily contaminated by mercury. The main source of contamination was a chlor-alkali plant, which was operating for 40 years since 1949. Previous studies showed that mercury concentrated in the sediment close to the plant is being dispersed throughout the bay by remobilization processes. Subsequent remobilization of mercury buried in the sediment may be harmful to various marine organisms. In order to assess the availability of remobilised mercury to marine organisms and to assess the health risk of consumption of shellfish species from the bay, a monitoring programme was initiated in September 1997. One thousand six hundred specimens of mussels (Mytilus galloprotincialis) of the same size and age, collected in a very clean area, were transferred to four stations in the bay 1.5 m above the sea bottom. Every month during the summer period and every second month in the winter period, 50 mussels were taken from each station for the analysis of mercury and monomethylmercury (MeHg) content. The results obtained during 1 year of biomonitoring suggest that the equilibrium concentration in transplanted mussels was established in a relative short period of time. The digestive gland is preferential organ for the accumulation of total mercury. Seasonal variation of mercury content both in the whole soft tissue, and organs (gills and digestive gland) was observed at all stations. However, methyl mercury is more concentrated in soft tissues than in digestive gland and gills, despite the fact that it is absorbed through these organs. The mercury levels in tissues were below accepted limits for human consumption. Obtained results showed that the implantation of mussels in an area containing sediment contaminated by mercury may be a good monitoring tool for the assessment of the availability of remobilised mercury to marine organisms. PMID- 11036978 TI - Total mercury in Pema perna mussels from Guanabara Bay--10 years later. AB - Guanabara Bay (GB) is an important estuary in regard to the productivity of the southeastern Brazilian coast. It is heavily polluted by oil, land runoff, and sewage, had large areas reclaimed, and its basin has been seriously deforested. The objective of the present work is to compare the total mercury levels in mussels from the bay, measured at the beginning and at the end of a 10-year interval (1988 and 1998). The commonly occurring species of bivalve in the bay is the mussel Perna perna. The total mercury content in mussels from GB has probably remained constant for the last 10 years (17.3-74.1 microg Hg kg(-1)), with possible isolated fluctuations attributed to occasional changes in water quality. Mussels have a good potential as biomonitors of water quality in GB. The need for the establishment of long-term biomonitoring programs is also commented. PMID- 11036979 TI - Detection of localized methylmercury contamination by use of the mussel adductor muscle in Minamata Bay and Kagoshima Bay, Japan. AB - Based on our previous finding that the concentrations of total mercury in mussel adductor muscle approximated those of methylmercury, we compared concentrations of total mercury in the adductor muscle of the mussel Mytilus galloprovincialis, collected from four sites around Minamata City from 1993 to 1995 and four sites in Kagoshima Bay from 1997 to 1998, to assess the level of localized methylmercury contamination. Though the input of mercury from the chemical plant had stopped by around 1970, concentrations of total mercury in the mussel adductor muscle were higher at two sites (26-121 ng/g, n = 135) near the main fallout of wastewater from the chemical plant in Minamata Bay than at the other sites, i.e. two sites 1-5 km from the former sites in Minamata City (6-28 ng/g, n = 52), and all sites in Kagoshima Bay (2-30 ng/g, n = 287). The localized methylmercury contamination around the chemical plant in Minamata Bay was documented also by our sensitive analysis of mercury concentrations in seawater and sediment samples. The survey of concentrations of total mercury in the mussel adductor muscle seems to be useful for monitoring the methylmercury contamination in coastal areas. PMID- 11036980 TI - Mercury methylation along a lake-forest transect in the Tapajos river floodplain, Brazilian Amazon: seasonal and vertical variations. AB - The seasonal and spatial variations of net methylmercury production in sediments, soils and other sites were evaluated by assays with 203Hg at different depths and locations along a lake-forest transect at lake Enseada Grande, Tapajos river. Soil and sediment samples were taken at the surface and at different depths up to 9 cm. Fresh samples and acidified controls (1-3 g dry wt.) were slurried with local water and incubated in the dark at 25-28 degrees C for 3 days with 0.5-1.6 microg Hg g(-1) (dry wt.) added as 203HgCl2. CH3 203Hg was extracted and measured in scintillation cocktail after acid leaching. Methylmercury production varied by orders of magnitude among sites and among sediment or soil layers. Seasonal variations were smaller than those with sample depth and location. In both seasons, MeHg formation in sediment and soil or flooded soil decreased with depth and was, in the top layers, one order of magnitude higher in the C-rich littoral macrophyte zone (2.3-8.9%) and flooded forest (3.2-4.5%) than in the center of the lake (0.2-0.56%). Similar MeHg production was found in slurried dry soils (dry season) and in soils already flooded for months. In the macrophyte zone soil (dry season), methylation was mainly associated with the thin Paspalum sp. rootlet layer. In the forest site, vertical variation in methylation was less pronounced in flooded than in dry soils and during the inundation the higher methylation rate was found in the flocculent sediment settled over the litter layer. The roots of floating Paspalum sp. were an important Hg methylation site, particularly those heavily colonized with periphyton (3.4-5.4%). Methylation in surface or near-bottom water was undetectable (< 3 x 10(-2)%) at all sites. Flooded forests and macrophyte mats are specific features of the Amazon and are important links between Hg inputs from natural and manmade sources and MeHg exposure of local populations through fish intake. PMID- 11036981 TI - Mercury net methylation in five tropical flood plain regions of Brazil: high in the root zone of floating macrophyte mats but low in surface sediments and flooded soils. AB - In aquatic systems, bottom sediments have often been considered as the main methylmercury (MeHg) production site. In tropical floodplain areas, however, floating meadows and flooded forests extend over large areas and can be important Hg methylating sites. We present here a cross-system comparison of the Hg net methylation capacity in surface sediments, flooded soils and roots of floating aquatic macrophytes, assayed by in situ incubation with 203Hg and extraction of formed Me203 Hg by acid leaching and toluene. The presence of mono-MeHg was confirmed by thin layer chromatography and other techniques. Study areas included floodplain lakes in the Amazon basin (Tapajos, Negro and Amazon rivers), the Pantanal floodplain (Paraguay river basin), freshwater coastal lagoons in Rio de Janeiro and oxbow lakes in the Mogi-Guacu river, Sao Paulo state. Different Hg levels were added in assays performed in 1994-1998, but great care was taken to standardise all other test parameters, to allow data comparisons. Net MeHg production was one order of magnitude higher (mean 13.8%, range 0.28-35) in the living or decomposing roots of floating or rooted macrophyte mats (Eichhornia azurea, E. crassipes, Paspalum sp., Eleocharis sellowiana, Salvinia sp., S. rotundifolia and Scirpus cubensis) than in the surface layer of underlying lake sediments (mean 0.6%, range 0.022-2.5). Methylation in flooded soils presented a wide range and was in some cases similar to the one found in macrophyte roots but usually much lower. In a Tapajos floodplain lake, natural concentrations of MeHg in soil and sediment cores taken along a lake-forest transect agreed well with data on net methylation potentials in the same samples. E. azurea, E. crassipes and Salvinia presented the highest methylation potentials, up to 113 times higher than in sediments. Methylation in E. azurea from six lakes of the Paraguay and Cuiaba rivers, high Pantanal, was determined in the 1998 dry and wet seasons and ranged from 1.8 to 35%. Methylation was lower in washed roots than in untreated roots of E. azurea and methylation in solids isolated from the roots, was higher than in sediments but lower than in untreated roots. This indicates that the methylation in roots zones occurs mainly in the root-associated solids. Floating meadows are sites of intense production of biomass and of highly bioavailable MeHg and appear to be an essential link of the MeHg cycle in tropical aquatic systems. PMID- 11036982 TI - Capacity of mercury volatilization by mer (from Escherichia coli) and glutathione S-transferase (from Schistosoma mansoni) genes cloned in Escherichia coli. AB - A study was carried out to evaluate the capacity for mercury volatilization by genetically engineered strains that express the mer and glutathione S-transferase genes from Escherichia coli and Schistosoma mansoni, respectively. This method enabled strains containing simultaneously mer and glutathione S-transferase genes to grow in high concentrations of mercuric chloride (30 microg/ml) and to volatilize part of the mercury (248 microg/g cell dry wt.) present in the culture medium, while strains bearing only a single gene, did not have the same behavior. Up to 70% of the total mercury of bacterial volatilization occurred in the first 4 h. Although the findings were preliminary, the genetically engineered strain containing simultaneously the mer and glutathione S-transferase genes show a great potential for bioremediation. It may be used in a closed system to remove by volatilization, and recover mercury (Hg0) from contaminated effluents, such as industrial effluent, for instance. PMID- 11036983 TI - The influence of humic substances on the speciation and bioavailability of dissolved mercury and methylmercury, measured as uptake by Chaoborus larvae and loss by volatilization. AB - The influence of dissolved humic substances (HS) on the bioavailability of dissolved inorganic and methyl mercury (Hg) was quantified by measuring the direct uptake of 203Hg in Chaoborus larvae using laboratory microcosms containing artificial freshwater. The animals were exposed individually in triplicate aquaria at 10 different concentrations of HS covering the whole range found in natural freshwaters (0-110 mg C l(-1)). Mercury-203 concentrations were monitored repeatedly in the same individuals and in their ambient water during up to 10 days. Near-steady state Hg concentrations in Chaoborus were usually reached within 5 days. The bioconcentration factor (BCF, direct uptake only) for the larvae in the absence of HS was 0.55+/-0.09 (S.E.) ml individual(-1) for inorganic Hg and 5.3+/-0.7 ml individual(-1) for methyl Hg, thus showing a 10 fold difference. Normalizing to the organic carbon content of the larvae yields a BCF(OC) in the absence of HS of 2.8+/-0.4 x 10(3) ml (gC)(-1) for inorganic Hg and 2.7+/-0.3 x 10(4) ml (gC)(-1) for methyl Hg. The uptake of both inorganic and methyl Hg decreased markedly with increasing concentration of HS. For inorganic Hg, the decrease in uptake was most pronounced at HS concentrations below 0.2 mg C l(-1). For methyl Hg, the relationship between uptake and log([HS]) was sigmoid, showing a reduction by > 90% when increasing HS concentrations from 1 to 50 mg C l(-1). Similar patterns were observed for losses of Hg from the water phase, mainly through volatilization. These results have implications for both the biouptake and the abiotic cycling of Hg in natural ecosystems and suggest that most dissolved inorganic Hg is bound to dissolved organic matter in most natural freshwaters, whereas dissolved methyl Hg is bound only in humic waters. Assuming that only free aqueous Hg is taken up by the organisms, the rather simple methodology employed here can be used for estimating distribution coefficients (K(OC)) for Hg between HS and water. In this study, the K(OC) values were 2.5+/-0.7 (S.E.) x 10(7) ml (gC)(-1) for inorganic Hg and 1.5+/-0.6 x 10(5) ml (gC)(-1) for methyl Hg. Values of similar magnitude were derived from observed losses of Hg from the water phase, supporting the assumption of an immobilization of both inorganic and methyl Hg by HS. The strong negative influence of dissolved HS on the bioavailability of both inorganic and methyl Hg in freshwater suggests that the high Hg levels often found in fish from humic lakes in the boreal forest zone cannot be explained alone by direct uptake of methyl Hg from the water phase into biota at low trophic levels. PMID- 11036984 TI - Photoreduction and evolution of mercury from seawater AB - The photoreduction of mercury (Hg2+ to Hg0) in natural seawater was investigated by means of a radiotracer (203Hg2+) solution exposed to natural and simulated sunlight. Different light regimes (dark, natural daylight, and a solar simulator), and dissolved organic carbon (DOC) levels from commercially available humic acids concentrations, were tested in the laboratory to evaluate the possibility of occurrence of the reaction in the environment. The natural seawater prepared accordingly to each experimental condition was continuously purged of the Hg0 formed, which was then re-oxidised in an acid trap and determined. The use of a solar simulator permitted the test of light intensity and wavelength dependence of the process under investigation. The reaction is dependent on the concentration of DOC in the experimental solution, increasing light intensity and decreasing wavelength. Reduction rates were in the range of 0.04-2.2% h(-1) for the DOC concentrations and light regimes tested. The process might have geochemical implications for the cycling of mercury around the air-sea interface. PMID- 11036985 TI - Electrochemically enhanced oxidation reactions in sandy soil polluted with mercury AB - For remediation of soils contaminated with heavy metals, the electrodialytic remediation (EDR) method is a highly relevant method, see e.g. Hansen et al. (Hansen HK, Ottosen LM, Kliem BK, Villumsen A. Electrodialytic remediation of soils polluted with Cu, Cr, Hg, Pb, and Zn. J Chem Tech Biotechnol 1997;70:67 73). During the process the heavy metals are transferred to the pore water in dissolved form or attached to colloids and move within the applied electric field. The method is found to be useful in many soil types, but has its strength in fine-grained soils. It is exactly in such soils that other remediation methods fail. Four cell experiments were made in order to investigate how relevant the method is for a more sandy soil and if it is suitable for non-ionic heavy metals such as elemental mercury. The duration was 27 days for two of the experiments and two experiments lasted 54 days, and the mercury within the soil was initially 1200-1900 mg kg(-1), of which 84% was elemental Hg. To monitor the process the pseudo-total mercury concentration was distinguished between elemental mercury and non-metallic mercury species by thermodesorption. During the electrodialytic treatment an increase of the content of non-metallic mercury occurred and a corresponding decrease of the content of elemental mercury which indicates a transformation of the latter species into any other non-metallic species. Generally, oxidation of Hg by dissolved oxygen in a solution is kinetically inhibited and thus quite slow. The redistribution of Hg was closely connected to a decrease of soil pH during the experiments. This corresponds very well to the thermodynamic calculations from which it was found that a decrease in the pH of the soil will result in an increase in the oxidation rate of elemental Hg. Results from this investigation show that the electrodialytic remediation method alone is not efficient in situations with sandy soils containing elemental mercury. As a solution for this problem it is suggested to add chloride to the soil system. Chloride would act as a complexing agent avoiding precipitation and enhancing the dissolution of precipitates as well as elemental mercury. PMID- 11036986 TI - Evaluating mercury transformation mechanisms in a laboratory--scale combustion system AB - Mercury speciation measurements during injections of 10 microg/m3 Hg0(g) into a 42-MJ/h combustion system containing gaseous O2-Ar- and O2-N2-rich mixtures indicate that 43 and 55% of the Hg (g) spike was transformed rapidly (< 0.1 s) to Hg2+X(g) within a refractory-lined heat exchanger where gas temperatures decrease from approximately 620 to 200 degrees C. O2(g) is the probable Hg0(g) oxidant (i.e. X = O2-). The apparent formation of HgO(g) involves a heterogeneous reaction with adsorbed Hg0 or O2 on refractory surfaces or a Hg0(g)-O2(g) reaction catalyzed by corundum (Al2O3) and/or rutile (TiO2) components of the refractory. The potential catalytic effects of Al2O3 and TiO2 on Hg0(g) oxidation were investigated by injecting Al2O3 and TiO2 powders into approximately 650 degrees C subbituminous coal (Powder River Basin, Montana, USA) combustion flue gas. On-line Hg0(g) and total mercury measurements indicate, however, that Al2O3 and TiO2 injections were ineffective in promoting the formation of additional Hg2+X(g). Apparently, either the chemically complex flue gas hindered the catalytic effects of Al2O3 and TiO2, or these compounds are simply not Hg0(g) oxidation catalysts. PMID- 11036987 TI - Characterization and speciation of mercury-bearing mine wastes using X-ray absorption spectroscopy. AB - Mining of mercury deposits located in the California Coast Range has resulted in the release of mercury to the local environment and water supplies. The solubility, transport, and potential bioavailability of mercury are controlled by its chemical speciation, which can be directly determined for samples with total mercury concentrations greater than 100 mg kg(-1) (ppm) using X-ray absorption spectroscopy (XAS). This technique has the additional benefits of being non destructive to the sample, element-specific, relatively sensitive at low concentrations, and requiring minimal sample preparation. In this study, Hg L(III)-edge extended X-ray absorption fine structure (EXAFS) spectra were collected for several mercury mine tailings (calcines) in the California Coast Range. Total mercury concentrations of samples analyzed ranged from 230 to 1060 ppm. Speciation data (mercury phases present and relative abundances) were obtained by comparing the spectra from heterogeneous, roasted (calcined) mine tailings samples with a spectral database of mercury minerals and sorbed mercury complexes. Speciation analyses were also conducted on known mixtures of pure mercury minerals in order to assess the quantitative accuracy of the technique. While some calcine samples were found to consist exclusively of mercuric sulfide, others contain additional, more soluble mercury phases, indicating a greater potential for the release of mercury into solution. Also, a correlation was observed between samples from hot-spring mercury deposits, in which chloride levels are elevated, and the presence of mercury-chloride species as detected by the speciation analysis. The speciation results demonstrate the ability of XAS to identify multiple mercury phases in a heterogeneous sample, with a quantitative accuracy of +/-25% for the mercury-containing phases considered. Use of this technique, in conjunction with standard microanalytical techniques such as X-ray diffraction and electron probe microanalysis, is beneficial in the prioritization and remediation of mercury-contaminated mine sites. PMID- 11036988 TI - Determination of total mercury in workers' urine in gold shops of Itaituba, Para State, Brazil. AB - Gold extraction and its commercialization in the Amazon region is mainly by rudimentary procedures. Therefore, during the process of extraction and recovery of this precious metal, large amounts of mercury vapors are thrown into the environment. This paper is an attempt to establish a correlation between the concentration of total mercury in the urine of workers at the gold shop in the Municipality of Itaituba, Para, and the information related to the lifestyles of each individual studied. Through statistical analysis, it was possible to divide the workers into three groups: people with normal mercury concentrations, [Hg] < or = 10 ppb, (29%); with concentrations at the biological limit of tolerance, [Hg] up to 50 ppb, (49%); and contaminated people, [Hg] > or = 50 ppb (22%). It may be concluded that fish consumption, time of alcohol consumption, number of amalgam fillings, as well as working hours, are important variables when evaluating mercurial contamination of people who are occupationally exposed to mercury vapors. PMID- 11036989 TI - The Northeast States and Eastern Canadian Provinces mercury study: a framework for action: summary of the Canadian chapter. AB - The objective of this paper is to give an overview of the findings of the Canadian chapter of the Northeast States and Eastern Canadian Provinces Mercury Study, which was conducted between 1995 and 1998. The Canadian chapter provided information on mercury emissions, sources and levels in air, water, biota and humans. Industry, governments and universities provided information for the Canadian chapter. The study showed that the Northeast States and Eastern Canadian Provinces (NES/ECP) is a region impacted by airborne mercury. Annual mercury emissions for the NES/ECP region are estimated to be approximately 19 t (12%) of the combined Canadian and US national anthropogenic mercury emissions of 155 t/year. 210Pb-dated lake sediment cores from Atlantic Canada showed a mercury enrichment factor of 2.5 for coastal sites with mercury increases starting in 1860. Regional mercury wet deposition for the NES/ECP region was 7-11 microg/m2/year. Provincial and federal fish health advisories have been updated in the ECP for children and women of child-bearing age limiting the consumption of freshwater fish, as well as fresh or frozen shark, tuna or swordfish. PMID- 11036990 TI - Developing consensus: mercury science and policy in the NAFTA countries (Canada, the United States and Mexico). AB - The international science community has recognized methylmercury in the aquatic food chain, as a potential environmental and human health risk. As a result, countries around the world have implemented a number of mercury management initiatives. The United States, Mexico and Canada in consultation with stakeholders are developing a trilateral North American Regional Action Plan (NARAP) on mercury. Through public involvement in the decision making process, the NARAP has offered opportunities for more transparency in transactions between governments, industry and stakeholders. In spite of the available scientific information, there are still important uncertainties associated with the mercury issue. These knowledge gaps include: the most appropriate methylmercury reference dose for sensitive groups; the percentage of the North American population that is at risk from methylmercury exposure; spatial and temporal mercury deposition patterns in each country; the link between mercury emissions, atmospheric deposition and methylmercury concentrations in fish; and the relative magnitude of contributions from natural and anthropogenic sources. PMID- 11036991 TI - Electrolytic treatment of mercury-loaded activated carbon from a gas cleaning system AB - This study aimed at extracting the adsorbed mercury from the mercury-loaded activated carbon so as to recycling both, the elemental mercury and the carbon, after being reactivated. The process used in this study was the electro-oxidation of the mercury in a reaction system where the loaded carbon is acting as an anode, during the electrolysis of brine, the electrolyte of the cell. PMID- 11036992 TI - Application of physico-chemical amendments for the counteraction of mercury pollution AB - In order to counteract mercury pollution, due to gold recovery practices, in situ strategies are required. In this study, some physico-chemical additions were tested, in different environmental compartments, to verify their potential to mitigate mercury contamination. Results indicate that the addition of sulfide reduces chemical methylation of Hg(II), but enhances the solubility of Hg0. The use of oxides, phosphate, and organic matter may be effective in the immobilization of Hg(II), depending upon Hg speciation. Calcium was effective in counteracting the solubility enhancement of Hg0 promoted by the presence of Aldrich humic acid. PMID- 11036993 TI - What is our standard of care? PMID- 11036994 TI - A comparison of shear bond strengths of three visible light-cured orthodontic adhesives. AB - The purpose of this study was to evaluate the shear bond strength and the site of bond failure for 2 visible light-cured composites (Transbond XT and Enlight) and a resin-modified glass ionomer cement (RMGIC; Fuji Ortho LC). Seventy-five extracted human premolars were collected and randomly divided into 3 test groups. Brackets were bonded to the teeth in each test group with the respective adhesive according to the manufacturers' instructions. Each specimen was debonded using an Instron Universal Testing Machine at a crosshead speed of 0.1 mm/min. The mode of bond failure was observed by using light microscopy. The results of this study demonstrated that the light-cured composites had a higher shear bond strength than the RMGIC. The adhesive-remnant scores were similar for the composites with the mean values at about 2, which indicates that more than half of the adhesive remained on the tooth. The RMGIC had a mean score of 3, which was significantly different from the composites and indicated that all of the adhesive remained on the tooth with a distinct impression of the bracket. PMID- 11036995 TI - Bonding of light-cured glass ionomer cement to polycarbonate resin treated with experimental primers. AB - The effect of experimental primers on the shear bond strength of polycarbonate composite resin with light-cured glass ionomer cements was investigated. Mixtures of methylmethacrylate (MMA) with the comonomers 2-hydroxyethyl methacrylate (HEMA), triethyleneglycol methacrylate (TEGDMA), and bisphenol-A glycidymethacrylate (bisGMA) were used as primers. Polycarbonate composite resin rods of circular cross section and plates were bonded, with and without precured and nonprecured primers, using 2 light-cured glass ionomer cements (commercially available [LC] and experimental [EX]). In addition, commercial polycarbonate composite resin brackets with precured 50% TEGDMA/MMA primer were bonded to etched human enamel with both cements. Shear bond strengths were measured. Results were compared by ANOVA and Scheffe's tests at P = .05. The 30% HEMA/MMA, 50% TEGDMA/ MMA, 10% bisGMA/MMA, and 30% bisGMA/MMA primers produced the higher shear bond strengths (9.5 to 20.8 MPa) with LC and EX to polycarbonate composite resin. The 50% TEGDMA/MMA primer was most effective in improving the shear bond strengths of both LC and EX. Precured 50% TEGDMA/ MMA primer on a commercial resin bracket was effective in providing good shear bond strength to enamel. PMID- 11036996 TI - Ongoing innovations in biomechanics and materials for the new millennium. AB - Material innovations are reviewed within the context of ongoing biomechanical developments that relate the critical contact angle of second-order angulation (theta c) to the overall resistance to sliding (RS). As a science in its embryonic stage of development, RS is partitioned into classical friction (FR), elastic binding (BI), and physical notching (NO). Both FR and BI are defined in terms of normal forces (N) and kinetic coefficients (mu k). The angulation at which NO occurs (theta z) is introduced as a second boundary condition to theta c. Given this scientific backdrop, material modifications are sought that reduce RS. Approaches include minimizing mu k or N within the context of FR and theta < theta c, as, for example, by surface modifications of arch wires and brackets or by engineering novel ligation materials. Stabilizing theta at theta approximately equal theta c should provide more efficient and effective sliding mechanics by developing innovative materials (eg, composites) in which stiffness (EI) varies without changing wire or bracket dimensions. Between the boundaries of theta c and theta z (ie, theta c < theta < theta z), BI may be reduced by decreasing EI or increasing interbracket distance (IBD), independent of whether a conventional or composite material is used. PMID- 11036997 TI - Response of headgear release mechanisms to nonaxial force application. AB - Safety products have been developed to help reduce the incidence of trauma caused by headgear. Previous studies have reported the characteristics of breakaway-type headgear release mechanisms with axial force application. Not all accidental releases are triggered by an axial force and it is necessary to understand the characteristics of these mechanisms with nonaxial force application. Thirteen headgear release mechanisms were tested as part of a complete headgear system. With the system attached to a plaster head and neck model a tensile force was applied to the system at 30 degrees to the sagittal plane at 2 rates. The force of activation at release and the distance traveled were determined and analyzed statistically. Force values ranged from 4.6 to 36.7 pounds and face bow travel before release ranged from 0.97 to 3.42 inches. No consistent pattern of rate dependence was observed. Several devices demonstrated the desirable combination of low force and face bow travel at release. PMID- 11036998 TI - Ultrasound imaging of condylar motion: a preliminary report. AB - Studies of condylar motion within the temporomandibular joint have been going on for some time. These studies have used techniques that included direct viewing via x-rays, magnetic resonance imaging, and arthroscopy, as well as indirect measurements using axiographs, kinesiographs, photocells, and light-emitting diodes. These viewing and measuring methods have important disadvantages and shortcomings. Recent advances in ultrasound technology, used extensively in medicine, have been adapted to dynamic imaging of the temporomandibular joint in the near-sagittal plane. Preliminary results strongly suggest that condylar motion is curvilinear throughout its range of motion. No evidence was seen to support the notion that condylar motion occurs about a fixed axis or point at any time during movement. Additional improvements in ultrasound technology may allow further definitive studies, and it may become usable in diagnosing temporomandibular dysfunction and disease states in the near future. PMID- 11036999 TI - Comparison of landmark identification in traditional versus computer-aided digital cephalometry. AB - The aim of this study was to assess landmark identification on digital images in comparison with those obtained from original radiographs. Ten cephalometric radiographs were selected randomly. Seven orthodontic residents identified 19 cephalometric landmarks on both the original radiographs and the digital images. To assess the concordance between landmarks identified on the original radiographs and on their digital counterparts, the x, y coordinates for each landmark in the 2 modalities were transformed with the identical references. The placement differences for 19 landmarks between 2 methods were calculated and their components in horizontal and vertical directions were analyzed respectively. Multivariate analysis of variance showed that the "cephalometric radiograph" and "landmark" variation had greater influence than that from "method" (landmark identification on digital / original radiograph). It was also noted that the differences of landmark identification between original radiographs and their digital counterparts were statistically significant. The landmarks with significant differences of horizontal component on the x-axis were Me, Gn, ANS, PNS, LIA. The differences were generally under 1 mm with the exception of Or, Me, PNS, LIA. The landmarks with significant differences of vertical component on the y-axis were Po, Or, Gn. The inter-observer error for each landmark in digital images was generally larger than that in the original radiographs. However, statistically significant differences of inter-observer errors between 2 modalities were only found for 4 of the 19 landmarks. These 4 landmarks, Po, Ar, ANS, and UM, should be scrutinized more carefully during potential applications of digital cephalometry. PMID- 11037000 TI - Perception of facial esthetics by native Chinese participants by using manipulated digital imagery techniques. AB - This investigation utilized a manipulated digital video imaging model to elicit profile facial esthetics preferences in a lay population of native Chinese participants from Beijing. A series of 4 distinct digitized distortions were constructed from an initial lateral cephalogram. These images represented skeletal or dental changes that differed by 2 standard deviations from the normative values for Chinese adults. Video morphing then created soft-tissue profiles. A series of nonparametric tests validated the digitized distortion model. The native Chinese participants in this sample found that the profile distortions most acceptable were the "flatter", or bimaxillary retrusive distortion, in the male stimulus face and the "anterior divergent", or maxillary deficiency, in the female stimulus face. PMID- 11037001 TI - Changes in position of the temporomandibular joint disc and condyle after disc repositioning appliance therapy: a functional examination and magnetic resonance imaging study. AB - Disc-repositioning splints are routinely used in the treatment of anteriorly displaced discs. The rationale of these appliances is to direct the mandibular condyle anteriorly in the glenoid fossa and to recapture the disc onto the condyle. The stability of disc recapture depends on reestablishment of the occlusion and the adaptive capabilities of the temporomandibular joint. It could therefore be suggested that treatment success is potentially higher in the active growth period. In this case report, partial disc recapture was observed on magnetic resonance images after application of a maxillary disc-repositioning appliance. Orthodontic treatment was applied for the retention of disc recapture. PMID- 11037002 TI - A precis of the proceedings of the first international orthodontic editor's symposium. PMID- 11037003 TI - The efficacy of ginkgo for elderly people with dementia and age-associated memory impairment: new results of a randomized clinical trial. AB - OBJECTIVES: To evaluate the efficacy, the dose-dependence, and the durability of the effect of the ginkgo biloba special extract EGb 761 (ginkgo) in older people with dementia or age-associated memory impairment. DESIGN: A 24-week, randomized, double-blind, placebo-controlled, parallel-group, multicenter trial. SETTING: Homes for the elderly in the southern part of the Netherlands. PARTICIPANTS: Older persons with dementia (either Alzheimer's dementia or vascular dementia; mild to moderate degree) or age-associated memory impairment (AAMI). 214 Participants were recruited from 39 homes for the elderly. INTERVENTION: The participants were allocated randomly to treatment with EGb 761 (2 tablets per day, total dosage either 240 (high dose) or 160 (usual dose) mg/day) or placebo (0 mg/d). The total intervention period was 24 weeks. After 12 weeks of treatment, the initial ginkgo users were randomized once again to either continued ginkgo treatment or placebo treatment. Initial placebo use was prolonged after 12 weeks. MEASUREMENTS: Outcomes were assessed after 12 and 24 weeks of intervention. Outcome measures included neuropsychological testing (trail-making speed (NAI-ZVT-G), digit memory span (NAI-ZN-G), and verbal learning (NAI-WL)), clinical assessment (presence and severity of geriatric symptoms (SCAG), depressive mood (GDS), self-perceived health and memory status (report marks)), and behavioral assessment (self-reported level of instrumental daily life activities). RESULTS: An intention-to-treat analysis showed no effect on each of the outcome measures for participants who were assigned to ginkgo (n = 79) compared with placebo (n = 44) for the entire 24-week period. After 12 weeks of treatment, the combined high dose and usual dose ginkgo groups (n = 166) performed slightly better with regard to self-reported activities of daily life but slightly worse with regard to self-perceived health status compared with the placebo group (n = 48). No beneficial effects of a higher dose or a prolonged duration of ginkgo treatment were found. We could not detect any subgroup that benefited from ginkgo. Ginkgo use was also not associated with the occurrence of (serious) adverse events. CONCLUSIONS: The results of our trial suggest that ginkgo is not effective as a treatment for older people with mild to moderate dementia or age-associated memory impairment. Our results contrast sharply with those of previous ginkgo trials. PMID- 11037004 TI - Dementia diagnostic guidelines: methodologies, results, and implementation costs. AB - OBJECTIVE: To facilitate the diagnostic process for dementia. Five guidelines and four consensus statements on specific diagnostic recommendations, specialist referral recommendations, and costs of recommended diagnostic procedures were compared and summarized. DATA SOURCES AND SELECTION: A MEDLINE search from 1984 to 1999 and queries to experts yielded 14 guidelines and consensus statements that addressed the diagnosis of dementia. Only nine documents which had national or international scopes were reviewed. METHODS: Comparisons were made on the specific diagnostic criteria for patient history, clinical examination, functional assessment, laboratory tests, neuroimaging, and other diagnostic tests, as well as specialist referral recommendations and costs for the recommended diagnostic procedures. The first three authors reviewed independently each document and completed a table on specific recommendations in each document. To settle disagreements about specific recommendations, they discussed them until they reached a consensus. To interpret the intent of vague statements, they used their best judgment. RESULTS: The documents differed in content, recommendations, and development methodology. They were based on either expert opinion or scientific evidence, or both. Although the nine documents were nearly unanimous in several recommendations, including assessing the presenting problem, taking a medical history, conducting physical and neurological examinations, and assessing the patient's mental and cognitive status, considerable differences in recommendations were common. Such differences led to large differentials in the estimated costs (range, $190 to $2,001) for recommended diagnostic assessments. CONCLUSIONS: A systematic approach to diagnostic recommendations for dementia may induce greater consistency among guidelines and consensus statements. The current approach leads to considerable variability in recommendations and estimated costs. PMID- 11037005 TI - The role of routine laboratory studies and neuroimaging in the diagnosis of dementia: a clinicopathological study. AB - OBJECTIVE: To determine the neuropathological diagnoses of longitudinally followed patients with potentially reversible causes of dementia and to examine the results of the "dementia work-up," especially neuroimaging, by comparison with the pathological diagnosis. DESIGN: A neuropathologic series of 61 consecutive patients, with review of clinical, laboratory, neuroimaging, and pathological results. RESULTS: Of the 61 patients, forty-eight (79%) had a clinical diagnosis of probable or possible Alzheimer's disease (AD). Compared with the pathological diagnosis, the sensitivity and specificity of the clinical diagnosis of AD were 96% and 79%, respectively. Of the 61 patients, 9 had abnormal laboratory tests, the correction of which did not improve the subsequent course. These patients were found to have AD8 and frontotemporal dementia on pathology. In two patients, neuroimaging was helpful in the clinical diagnoses of frontotemporal dementia and progressive supranuclear palsy (PSP). Neuroimaging revealed cerebrovascular disease in 18 patients, only two of whom were suspected clinically. Pathology confirmed AD in 17 and PSP in 1 of these patients. Sensitivity and specificity for the clinical diagnosis of cerebrovascular disease in comparison with pathology were 6% and 98%, respectively. With the added information from neuroimaging, that sensitivity increased to 59% and specificity decreased to 81%. CONCLUSIONS: All cases with abnormal laboratory or neuroimaging results had AD or some other neurodegenerative disease on pathology. The "dementia work-up" did not reveal any reversible causes for dementia in this group of patients. Neuroimaging may have a role, especially in the diagnosis of possible AD with concomitant cerebrovascular disease. PMID- 11037006 TI - Preventing the spread of vancomycin-resistant enterococci in a long-term care facility. AB - OBJECTIVES: To test the hypothesis that infection control practices can prevent the spread of vancomycin-resistant enterococci (VRE) to residents of a long-term care facility (LCF) from an affiliated acute care facility with a high endemic rate of colonization. DESIGN: Point prevalence study of the rate of rectal colonization. SETTING: A state-supported veterans nursing home and an acute care veterans hospital. PARTICIPANTS: Residents in a state veterans home. INTERVENTIONS: Identification of patients with rectal colonization by VRE before transfer to the state veterans home, contact isolation for colonized veterans, use of oral bacitracin to eliminate colonization. MEASUREMENTS: Rectal swab and culture for VRE, review of clinical records and recording of presumptive risk factors for VRE colonization. The risk factors were age, gender, length of stay at nursing home, treatment with vancomycin or oral antibiotics, prior hospitalization at the acute care facility during the prior year, use of indwelling urethral catheters, presence of diarrhea, and fecal or urinary incontinence. RESULTS: Sixty-nine of 200 residents were cultured in the first study (1996) and 130 of 230 residents were cultured in the second study (1998). Residents who consented to culture differed from those who did not only with regards to gender (2 vs 7, P = .012). In neither study were any residents found to be colonized with VRE who had not already been identified as positive on admission. CONCLUSIONS: Adherence to infection control practices by the patient care staff of the LTCF was associated with the absence of transmission of VRE colonization among its residents. The presence of rectal colonization with VRE in an acute care patient should not be a barrier to acceptance in a nursing home. PMID- 11037007 TI - A serious outbreak of parainfluenza type 3 on a nursing unit. AB - OBJECTIVE: To report a serious outbreak of respiratory illness in a nursing home, with isolation of parainfluenza type 3 in four cases. DESIGN: Viral respiratory cultures from a sample of symptomatic residents, and retrospective chart review. SETTING: A 50-bed nursing unit/floor in a skilled nursing facility. PARTICIPANTS: All residents of the nursing unit. MEASUREMENTS: Respiratory viral cultures and clinical chart review. RESULTS: Twenty-five of 49 residents developed new respiratory symptoms between September 2 and September 25, 1999. Ten cases (40%) had a tympanic temperature of 100 degrees F or greater. Eighteen (72%) had a chest X-ray with 11 (44%) new infiltrates. Sixteen (64%) were treated with antibiotics. Three cases were hospitalized and four died (16%) within 1 to 9 days after onset of symptoms. Four of 10 viral cultures yielded parainfluenza type 3. CONCLUSIONS: Parainfluenza type 3 may cause outbreaks complicated by pneumonia and fatal outcome. Clinicians should consider uniform secretion precautions to contain all viral URIs in nursing homes. PMID- 11037008 TI - Physician orders for life-sustaining treatment (POLST): outcomes in a PACE program. Program of All-Inclusive Care for the Elderly. AB - OBJECTIVES: To evaluate whether terminal care was consistent with Physician Orders for Life-Sustaining Treatment (POLST), a preprinted and signed doctor's order specifying treatment instructions in the event of serious illness for CPR, levels of medical intervention, antibiotics, IV fluids, and feeding tubes. DESIGN: Retrospective chart review. SETTING: ElderPlace, a Program of All Inclusive Care for the Elderly (PACE) site in Portland, Oregon. PARTICIPANTS: All ElderPlace participants who died in 1997 were eligible (n = 58). Reasons for exclusion were no POLST (1), missing POLST (1), and insufficient documentation of care (2). MEASUREMENTS: POLST instructions for each participant and whether or not each of the treatments addressed by the POLST was administered in the final 2 weeks of life. RESULTS: The POLST specified "do not resuscitate" for 50 participants (93%); CPR use was consistent with these instructions for 49 participants (91%). "Comfort care" was the designated level of medical intervention in 13 cases, "limited interventions" in 18, "advanced interventions" in 18, and "full interventions" in 5. Interventions administered were at the level specified in 25 cases (46%); at a less invasive level in 18 (33%), and at a more invasive level in 11 (20%). Antibiotic administration was consistent with POLST instructions for 86% of 28 subjects who had infections in the last 2 weeks of life, and less invasive for 14%. Care matched POLST instructions in 84% of cases for IV fluids and 94% for feeding tubes. CONCLUSIONS: POLST completion in ElderPlace exceeds reported advance directive rates. Care matched POLST instructions for CPR, antibiotics, IV fluids, and feeding tubes more consistently than previously reported for advance directive instructions. Medical intervention level was consistent with POLST instructions for less than half the participants, however. We conclude that the POLST is effective for limiting the use of some life-sustaining interventions, but that the factors that lead physicians to deviate from patients' stated preferences merit further investigation. PMID- 11037009 TI - Mammography use, breast cancer stage at diagnosis, and survival among older women. AB - BACKGROUND: Women age 65 years and older account for most newly diagnosed breast cancers and deaths from breast cancer. Yet, older women are least likely to undergo mammography, perhaps because mammography's value is less well demonstrated in older women. OBJECTIVE: To investigate the relationship between prior mammography use, cancer stage at diagnosis, and breast cancer mortality among older women with breast cancer. DESIGN: Retrospective cohort study using the Linked Medicare-Tumor Registry Database. SETTING: Population-based data from three geographic areas included in the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) program. PARTICIPANTS: Women aged 67 and older diagnosed with a first primary breast cancer, from 1987 to 1993, residing in Connecticut, metropolitan Atlanta, Georgia, or Seattle-Puget Sound, Washington. MEASUREMENTS: Medicare claims were reviewed and women were classified according to their mammography use during the 2 years before diagnosis: nonusers (no prior mammograms), regular users (at least two mammograms at least 10 months apart), or peri-diagnosis users (only mammogram(s) within 3 months before diagnosis). Mammography utilization was linked with SEER data to determine stage at diagnosis and cause of death. Our main outcome variables were (1) stage at diagnosis, classified as early (in situ/Stage I) or late (Stage II or greater), and (2) breast cancer mortality, measured from diagnosis until death from breast cancer or end of the follow-up period (December 31, 1994). RESULTS: Older women who were nonusers of mammography were diagnosed with breast cancer at Stage II or greater more often than regular users (adjusted odds ratio (OR), 3.12; 95% confidence interval (CI), 2.74-3.58). This association was present within each age group studied. Nonusers of mammography were at significantly greater risk of dying from their breast cancer than regular users for all women (adjusted hazard ratio (HR), 3.38; 95% CI, 2.65-4.32) and for women within each age group. Even assuming a lead time of 1.25 years, nonusers of mammography continued to be at increased risk of dying from breast cancer. Our findings remained significant for all women and for the two youngest age groups (67-74 years, 75-85 years), although the benefit was no longer statistically significant for the oldest women (85 years and older). CONCLUSIONS: Older women who undergo regular mammography are diagnosed with an earlier stage of disease and are less likely to die from their disease. These data support the use of regular mammography in older women and suggest that mammography can reduce breast cancer mortality in older women, even for women age 85 and older. PMID- 11037010 TI - Sleep problems as a risk factor for falls in a sample of community-dwelling adults aged 64-99 years. AB - OBJECTIVES: The purpose of this study was to determine if reported nighttime sleep problems and daytime sleepiness were associated with reported falling during the previous 12 months in a representatively sampled older adult population. DESIGN: Random-digit dial telephone survey. SETTING: Representatively sampled older adult population living in northern California. PARTICIPANTS: Participants were 971 women and 555 men, aged 64 to 99 years. MEASUREMENTS: Twenty-minute telephone interview adapted from the National Health Interview Survey. RESULTS: Two hundred and eighty-four participants reported falling during the previous 12 months (19% of the sample). Significantly more women fell than men (20% and 14%, respectively, P < .001). The following variables were significant risk factors for falling in univariate analyses: female gender, being unmarried, living alone, income less than $15,000 per year, difficulty walking, having more than one chronic medical condition, history of cardiovascular disease, hypertension, arthritis, sensory impairment, psychological difficulties, and nighttime sleep problems. All of the nighttime sleep problem variables remained significant risk factors for falling after controlling for other risk factors for falling. CONCLUSIONS: The results provide support for an independent association between reported sleep problems and falls in an older population. One of the implications of these data is that behavioral research focusing on the effectiveness of insomnia treatment in old age should not only examine typical sleep-related outcomes (e.g., total time asleep, number of awakenings) but also the occurrence of falls as well. PMID- 11037011 TI - Factors related to sleep disturbance in older adults experiencing knee pain or knee pain with radiographic evidence of knee osteoarthritis. AB - OBJECTIVES: To describe the types and frequencies of sleep complaints and the biopsychosocial factors associated with sleep disturbance in a large community sample of older adults experiencing knee pain or knee pain with radiographic evidence of knee osteoarthritis (OA). DESIGN: Baseline analyses of an observational prospective study. SETTING AND PARTICIPANTS: Participants were 429 men and women aged 65 years and older experiencing knee pain or knee pain with radiographic evidence of OA enrolled in the Observational Arthritis Study in Seniors (OASIS). MEASUREMENTS: Demographic variables (age, gender, ethnicity, education), health (X-rays of knee rated for OA severity, medical conditions, medication use, smoking status, body mass index, self-rated health), physical functioning (self-rated physical functioning, physical performance), knee pain, and psychosocial functioning (social support, depression) were measured. RESULTS: Problems with sleep onset, sleep maintenance, and early morning awakenings occurred at least weekly among 31%, 81%, and 51% of participants, respectively. Bivariate correlates of greater sleep disturbance in those with OA were less education, cardiovascular disease, more arthritic joints, poorer self-rated health, poorer physical functioning, poorer physical performance, knee pain, depression, and less social support. In regression analyses, each set of variables representing the domains of health, physical functioning, pain, and psychosocial functioning contributed to the prediction of sleep disturbance beyond the demographic set. Finally, in a simultaneous model, white race (trend, P = .06), poorer self-rated health, poorer physical functioning, and depressive symptoms were predictive of sleep disturbance. CONCLUSIONS: Sleep disturbance is common in older adults experiencing knee pain or knee pain with radiographic evidence of OA and is best understood through the consideration of demographic, physical health, physical functioning, pain, and psychosocial variables. Interventions that take into account the multidetermined nature of sleep disturbance in knee pain or knee OA are most likely to be successful. PMID- 11037012 TI - Prevalence, incidence, and risk factors associated with hip fractures in community-dwelling older Mexican Americans: results of the Hispanic EPESE study. Establish Population for the Epidemiologic Study for the Elderly. AB - OBJECTIVES: To determine the rates and risk factors associated with hip fractures in the community-dwelling older Mexican-American population. DESIGN: A prospective survey of a regional probability sample of older Mexican Americans aged 65 and over. SETTING: The 1993-1996 Hispanic Established Population for the Epidemiologic Study of the Elderly (H-EPESE), a probability sample of noninstitutionalized Mexican Americans, aged 65 and over, living in the Southwestern states of Texas, New Mexico, Colorado, Arizona, and California. PARTICIPANTS: In 1993-1994 and in 1995-1996, 2895 persons, aged 65 and over, considered Mexican American, were selected at baseline as a weighted probability sample. Sample weights were used to extrapolate to the estimated 498,176 older Mexican Americans residing in the Southwest US. MEASUREMENTS: Self-reported hip fracture and functional measures by in home interviews. RESULTS: Hip fracture prevalence was 4.0% at baseline. The overall incidence of hip fractures for women was 9.1 fractures/1000 person-years. The incidence rate for men was 4.8 fractures/1000 person-years. Extrapolation from these data to the entire older Mexican American population indicated that approximately 5162 new fractures occurred in the population during the 2 year study period. In women, hip fractures were associated independently with advanced age, not being married/living alone, having had a stroke, limitations with activities of daily living and instrumental activities of daily living. In men, only the latter limitations were associated independently with hip fracture. CONCLUSIONS: This study indicates that older Mexican American people may have hip fracture incidence rates that place them at highest risk among the Hispanic subgroups. In light of a sparse literature on this population, the fracture estimates derived from this work contributes to our understanding of the true fracture estimates in this population. Based on the extrapolated population rates, hip fracture in this population is a significant public health problem. Adequate preventive measures need to be implemented in this growing US population. PMID- 11037013 TI - Expert physician recommendations and current practice patterns for evaluating and treating men with osteoporotic hip fracture. AB - OBJECTIVES: To develop recommendations for the evaluation and the treatment of men with osteoporotic hip fracture from expert publications in the field of male osteoporosis, and to define the current practice patterns in a tertiary care VA Medical Center in Durham, North Carolina. DESIGN: Survey research; a retrospective cohort study. SETTING: Tertiary care VA Medical Center in Durham, North Carolina. PARTICIPANTS: (1) US physicians who published on the subject of male osteoporosis in the peer-reviewed literature between 1993 and 1997 identified by MEDLINE database search. (2) All 119 men admitted to the Durham VA Medical Center with ICD9 code for hip fracture between 1994 and 1998. OUTCOME MEASURES: (1) Osteoporosis evaluation and treatment recommendations of published physicians obtained by survey instrument. (2) Actual osteoporosis evaluation completed and therapy prescribed during index hospitalization in a cohort of men with hip fractures, determined by chart and database review. RESULTS: (1) Forty three physician-researchers were surveyed with an 84% response rate. For an osteoporosis evaluation, 89% of respondents recommended measuring serum testosterone, 85% serum calcium, 75% 25-OH vitamin D levels, 73% myeloma screen, and 61% serum thyroid-stimulating hormone (TSH). Dual Energy X-ray Absorptiometry would be obtained by 92%. More than 70% recommended calcium, vitamin D, and bisphosphonates for men with a normal metabolic evaluation, and 60% suggested weight-bearing exercise. (2) In the cohort of men admitted with hip fractures, 50% had a serum calcium level and 3% had a serum TSH level measured. Vitamin D was prescribed to 25% of patients in the form of a multivitamin, and 4% received calcium. There was no bisphosphonate, testosterone, or calcitonin use. CONCLUSIONS: Physicians who have published on osteoporosis recommended metabolic evaluation and osteoporosis therapy after hip fracture. Only minimal evaluation and treatment occurred in a cohort of men with osteoporotic hip fractures. PMID- 11037014 TI - Physical activity, functional limitations, and disability in older adults. AB - OBJECTIVES: To explore initially how low levels of physical activity influence lower body functional limitations in participants of the Longitudinal Study of Aging. Changes in functional limitations are used subsequently to predict transitions in the activities of daily living/instrumental activities of daily living (ADL/IADL) disability, thus investigating a potential pathway for how physical activity may delay the onset of ADL/IADL disability and, thus, prolong independent living. DESIGN: Analysis of a complex sample survey of US civilian, noninstitutionalized population aged 70 years and older in 1984, with repeated interviews in 1986, 1988, and 1990. SETTING AND PARTICIPANTS: Analyses concentrated on 5151 men and women targeted for interview at all four LSOA interviews. MEASUREMENTS: Characteristics used in analyses: gender, age, level of physical activity, comorbid conditions including the presence of hypertension, diabetes, arthritis, and atherosclerotic heart disease, levels of functional limitations, and ADL/IADL disability. RESULTS: Transitional models provide evidence that older adults who have varying levels of disability and who report at least a minimal level of physical activity experience a slower progression in functional limitations (OR = .45, P < .001 for severe vs less severe limitations). This low level of physical activity, through its influence on changes in functional limitations, is shown to slow the progression of ADL/IADL disability. CONCLUSIONS: Results from analyses provide supporting evidence that functional limitations can mediate the effect that physical activity has on ADL/IADL disability. These results contribute further to the increasing data that seem to suggest that physical activity can reduce the progression of disability in older adults. PMID- 11037015 TI - Moderate alcohol consumption and hearing loss: a protective effect. AB - OBJECTIVE: To determine if moderate alcohol consumption is associated inversely with hearing loss in a large population based study of older adults. DESIGN: Cross-sectional population based cohort study. Data are from the 1993-1995 examinations for the population based Epidemiology of Hearing Loss Study (EHLS) (n = 3571) and the Beaver Dam Eye Study (BDES) (n = 3722). SETTING: Midwestern community of Beaver Dam, Wisconsin. PARTICIPANTS: Residents of Beaver Dam aged 43 to 84 in 1987-1988 were eligible for the BDES (examinations in 1988-1990 and 1993 1995). During 1993-1995, this same cohort was eligible to participate in the baseline examination for the EHLS. MEASUREMENTS: Hearing thresholds were measured by pure tone air and bone conduction audiometry (250-8000 Hz.). History of alcohol consumption in the past year, heavy drinking (ever), medical history, occupation, noise exposure, and other lifestyle factors were ascertained by a questionnaire that was administered as an interview. RESULTS: In multiple logistic regression analyses controlling for potential confounders, moderate alcohol consumption (>140 grams/week) was inversely associated with hearing loss (PTA(.5,1,2,4 > 25 dB HL); odds ratio [OR] = .71, 95% confidence interval [CI] = .52, .97; where PTA is pure tone average). A similar association was found for moderate hearing loss (PTA(.5,1,2,4 > 40 dB HL); OR = 0.49, 95% CI = 0.32, 0.74). Alcohol consumption was associated inversely with the odds of having a low frequency hearing loss (OR = 0.61) or a high frequency hearing loss (OR = 0.60). These findings did not vary significantly by age or gender. There was an increase in the odds of having a high frequency hearing loss (OR = 1.35, 95% CI = 1.04, 1.75), in those with a history of heavy drinking (> or =4 drinks/day). Including cardiovascular disease or its related factors did not significantly attenuate the protective effect. CONCLUSIONS: There is evidence of a modest protective association of alcohol consumption and hearing loss in these cross-sectional data. This finding is in agreement with a small body of evidence suggesting that hearing loss is not an inevitable component of the aging process. PMID- 11037016 TI - Impact of depressive symptoms on hospitalization risk in community-dwelling older persons. AB - OBJECTIVE: To determine whether depressive symptoms in older adults are associated with an increased risk for hospitalization. DESIGN: A 6 month cohort study. SETTING: Five counties in the northern Piedmont of North Carolina from the Duke University site of the Established Populations for Epidemiological Studies of the Elderly project. PARTICIPANTS: The sample included 3486 community-dwelling adults, aged 65 and older. MEASUREMENTS: Crude risk ratios for the effect of depressive symptoms on 6 month risk for hospitalization were calculated, followed by a multivariable analysis controlling for demographics and health status. RESULTS: Three hundred participants were hospitalized during the 6 month follow up period. The crude risk ratio for the effect of depressive symptoms on hospitalization was 1.95 (95% CI = 1.47-2.58). Subgroup analysis showed significant positive risk ratios for men aged 65 to 74 and > or =75, and women aged 65 to 74. After a multivariable analysis, however, these associations remained significant only among men > or =75 (RR = 3.43; 95% CI = 1.33-8.86). CONCLUSIONS: Depressive symptoms were independently associated with a more than threefold increased risk for hospitalization among men aged > or =75. This result reflects differences in the effects of depressive symptoms across age and gender groups, and emphasizes that symptoms of depression influence overall health and medical utilization among, at the very least, the oldest subset of men. PMID- 11037017 TI - Cognitive decline is associated with systemic oxidative stress: the EVA study. Etude du Vieillissement Arteriel. AB - OBJECTIVE: To determine whether systemic oxidative stress status is associated with cognitive decline. DESIGN: A longitudinal population-based study. SETTING: A cohort study of older subjects in Nantes, France. PARTICIPANTS: A total of 1166 high cognitive functioning subjects aged 60 to 70 in the Etude du Vieillissement Arteriel (EVA) cohort with a 4 year follow-up. MEASUREMENTS: Subjects completed a baseline interview and a global cognitive test (Mini-Mental Status Examination (MMSE)). Blood samples were obtained at baseline to determine plasma levels of selenium, carotenoids, thiobarbituric acid reactant substances (TBARS), an indicator of lipoperoxidation, and red blood cell vitamin E. Risk of cognitive decline, defined as a loss of 3 points in MMSE score between baseline and the 4 year follow-up, was assessed by oxidative stress level. RESULTS: Subjects with the highest levels of TBARS show an increased risk of cognitive decline (adjusted odds ratio (OR) = 2.25; confidence interval (CI) 95% = 1.26-4.02). This result is reinforced in the lower antioxidant status subgroup. Subjects with low levels of selenium have an increased risk of cognitive decline (OR = 1.58; CI 95% = 1.08 2.31) after adjustment for various confounding factors. CONCLUSIONS: These results suggest that increased levels of oxidative stress and/or antioxidant deficiencies may pose risk factors for cognitive decline. The direct implication of oxidative stress in vascular and neurodegenerative mechanisms that lead to cognitive impairment should be further explored. PMID- 11037018 TI - Outcome of nursing home-acquired pneumonia: derivation and application of a practical model to predict 30 day mortality. AB - OBJECTIVES: To derive a prediction model of 30 day mortality for nursing home acquired pneumonia (NHAP) based on factors that can be readily identified by nursing home staff at the time of diagnosis and to apply the model to management issues related to NHAP including clarifying the importance of prepneumonia functional status as a predictor of outcome of NHAP. DESIGN: This was a retrospective chart review of 378 episodes of NHAP treated in the nursing home or hospital during two periods: November 1997 to April 1998 and November 1998 to April 1999. SETTING: Eleven nursing homes in the greater Buffalo, NY region. PARTICIPANTS: Nursing home residents with radiographically proven pneumonia who had at least one of the following signs/symptoms: cough, fever, purulent sputum, respiratory rate > or =25 breaths/minute, localized auscultatory findings, or pleuritic pain. MEASUREMENTS: Status (alive or dead) of each resident at 30 days (30 day mortality) after diagnosis of NHAP was the dependent variable. Factors predicting 30 day mortality were identified by logistic regression analysis. A scoring system was developed based on the results of the logistic model. Each episode of NHAP in the derivation cohort was scored using the model and the cohort was stratified by the model score into six categories or risk for mortality (0-5). The predictability of the model in the derivation cohort was measured using receiver operator characteristics curve analysis. RESULTS: Of 378 episodes of NHAP, 74% were treated initially in the nursing home and 26% were hospitalized initially for treatment. The overall 30 day mortality was 21.4%; however, the mortality rate was significantly higher for those treated initially in the hospital (29.6% vs 16.6%; P = .012). Logistic regression analysis identified four predictors of 30 day mortality: (1) respiratory rate >30 breaths/minute (2 points), (2) pulse > 125 beats/minute (1 point), (3) altered mental status (1 point), and (4) a history of dementia (1 point). Applying the scoring system to each episode in the derivation cohort demonstrated increasing mortality with increasing score. The c statistic for the model in the derivation cohort was .74. Based on the severity of NHAP, model episodes treated initially in the hospital were more acutely ill than those who were treated initially in the nursing home, and episodes treated with a parenteral antibiotic in the nursing home were more acutely ill than those who were treated with an oral agent. Functional status was not a predictor of 30 day mortality although there was a trend of higher mortality in the most dependent group (P = .065). The severity of NHAP model was able to define low and high risk mortality groups within a functional status category. CONCLUSIONS: A severity of NHAP model was derived from a large cohort of episodes in multiple facilities. The model had reasonable discriminatory power in the derivation cohort. The model may aid clinicians in making treatment decisions in the nursing home setting and in making hospitalization decisions. Although prepneumonia functional status provides a reasonable estimate of NHAP severity and prognosis, the severity of NHAP model permitted further refinement of these estimates. The severity of NHAP model requires validation before it can be recommended for general use. PMID- 11037019 TI - Relationships between nutritional markers and the mini-nutritional assessment in 155 older persons. AB - OBJECTIVE: To investigate the relationships between nutritional status measured by a comprehensive nutritional assessment including anthropometric measurements, nutritional biological markers, evaluation of dietary intake, and the Mini Nutritional Assessment (MNA) nutrition screening tool. DESIGN: A prospective study. PARTICIPANTS: One hundred fifty-five older subjects (53 men and 102 women; mean age = 78 years; range = 56-97 years). These participants were hospitalized in a geriatric evaluation unit (n = 105) or free living in the community (n = 50). MEASUREMENT: Weight, height, knee height, midarm and calf circumferences, triceps and subscapular skinfolds, albumin, transthyretin (prealbumin), transferrin, ceruloplasmin, C-reactive protein, alpha1-acid glycoprotein, cholesterol, vitamins A, D, E, B1, B2, B6, B12, folate, copper, zinc, a 3 day food record combined with a food-frequency questionnaire; the MNA nutritional screening. RESULTS: The MNA scores have been found to be significantly correlated to nutritional intake (P < .05 for energy, carbohydrates, fiber, calcium, vitamin D, iron, vitamin B6, and vitamin C), anthropometric and biological nutritional parameters (P < .001 for albumin, transthyretin, transferrin, cholesterol, retinol, alpha-tocopherol, 25-OH cholecalciferol zinc). An MNA score between 17 and 23.5 can identify those persons with mild malnutrition in which nutrition intervention may be effective. CONCLUSIONS: The MNA is a practical, noninvasive, and cost-effective instrument allowing for rapid nutritional evaluation and effective intervention in frail older persons. PMID- 11037020 TI - Influenza outbreak detection and control measures in nursing homes in the United States. AB - OBJECTIVE: To evaluate the use of influenza vaccine, rapid influenza testing, and influenza antiviral medication in nursing homes in the US to prevent and control outbreaks. METHODS: Survey questionnaires were sent to 1017 randomly selected nursing homes in nine states. Information was collected on influenza prevention, detection and control practices, and on outbreaks during three influenza seasons (1995-1998). RESULTS: The survey response rate was 78%. Influenza vaccine was offered to residents and staff by 99% and 86%, respectively, of nursing homes. Among nursing homes offering the influenza vaccine, the average vaccination rate was 83% for residents and 46% for staff. Sixty-seven percent of the nursing homes reported having access to laboratories with rapid antigen testing capabilities, and 19% reported having a written policy for the use of influenza antiviral medications for outbreak control. Nursing homes from New York, where organized education programs on influenza detection and control have been conducted for many years, were more likely to have reported a suspected or laboratory-confirmed influenza outbreak (51% vs 10%, P = .01), to have access to rapid antigen testing for influenza (92% vs 63%, P = .01), and to use antivirals for prophylaxis and treatment of influenza A for their nursing home residents (94% vs 55%, P = .01) compared with nursing homes from the other eight states. CONCLUSIONS: Influenza outbreaks among nursing home residents can lead to substantial morbidity and mortality when prevention measures are not rapidly instituted. However, many nursing homes in this survey were neither prepared to detect nor to control influenza A outbreaks. Targeted, sustained educational efforts can improve the detection and control of outbreaks in nursing homes. PMID- 11037021 TI - Nonspecific presentation of pneumonia in hospitalized older people: age effect or dementia? AB - OBJECTIVES: Older adults, when presenting with pneumonia, are often thought to present with nonspecific symptoms instead of more suggestive symptom(s). However, studies designed to determine whether age is associated with nonspecific presentations have yielded contradictory results. Many studies have not distinguished between the effects of preexisting cognitive impairment that results from dementia and the effects of age. The aim of this study is to determine whether there are significant differences in the presentation of pneumonia in demented versus nondemented patients across two age groups. We hypothesized that the nonspecific presentation of pneumonia in older people is due to dementia rather than to chronological age. DESIGN: We compared retrospectively nonspecific (weakness, decreased appetite, urinary incontinence, falls, and delirium) and specific (cough, sputum production, dyspnea, and chest pain) symptoms of pneumonia in 148 hospitalized adult subjects from two urban, general medical teaching hospitals. RESULTS: When the subjects with dementia were included in the analysis, two (falls and delirium) of the five nonspecific symptoms were associated with older age and one other symptom (weakness) showed a trend toward statistical significance. However, when we excluded the demented subjects, nonspecific presenting symptoms were similar in old and young adults with the exception of an increased frequency of delirium on presentation. Similarly, when demented subjects were excluded, we found a stronger association of younger age with the classic specific symptoms than were seen when the demented subjects were included. CONCLUSIONS: We conclude that age differences in the presentation of pneumonia are largely due to the presence of dementia. PMID- 11037022 TI - Nocturnal polyuria in older people: pathophysiology and clinical implications. AB - OBJECTIVES: To review the physiological changes of aging which affect the systems involved in urine formation and to consider how these changes interact with changes in bladder function, thereby leading to the onset of nocturnal polyuria with associated urinary frequency, nocturia, and incontinence. Based on this information, data are presented on the effectiveness of pharmacological interventions which reduce the rate of urine formation and, thus, can be of benefit in reducing symptoms, especially during the nighttime. METHODS: Peer reviewed journal articles were identified by MEDLINE Search and by review of the literature. CONCLUSIONS: As a consequence of age-associated diminished renal concentrating capacity, diminished sodium conserving ability, loss of the circadian rhythm of antidiuretic hormone secretion, decreased secretion of renin angiotensin-aldosterone, and increased secretion of atrial natriuretic hormone, there is an age-related alteration in the circadian rhythm of water excretion leading to increased nighttime urine production in older people. The interaction of nocturnal polyuria with age-related diminution in functional bladder volume and detrusor instability results in the symptoms of urinary frequency, nocturia and, in some persons, incontinence. The additional impact of Alzheimer's disease on these physiological and aging changes, as well as on a diminished perception of bladder fullness, leads to an even greater risk of urinary incontinence in these patients. Treatment of nocturnal polyuria with the antidiuretic hormone analog, DDAVP (desmopressin), can result in decreased nocturnal urine production with improvement in symptoms of frequency, nocturia, and incontinence. PMID- 11037023 TI - Health care utilization by old-old long-term care facility residents: how do Medicare fee-for-service and capitation rates compare? AB - OBJECTIVE: To describe the healthcare utilization of a long-term care population receiving primary and specialty care in a closed system and to compare Medicare fee-for-service (FFS) reimbursement with the amount that would have been paid under capitation for these services. SETTING: A life care community in California composed of two facilities, both having residential care and nursing facility (NF) beds. PARTICIPANTS: Residents (n = 700) living in the community between September 1995 and February 1996. METHODS: Data on Medicare Part A and Part B reimbursements were gathered from billing records for hospitalizations, based on diagnostic related group payments, primary and specialty care visits, various procedures, diagnostic tests, and therapeutic services. These data were compared with what the facility, in collaboration with the providers and an affiliated hospital, would have received under Medicare capitated rates at that time. RESULTS: Annually, residents averaged 16.3 primary care visits, 7.7 specialist visits, and 3453 hospital days per thousand. Nursing facility residents received significantly more primary care than did those in residential care. Total Medicare Part A and B payments per resident per month averaged $558. The monthly capitation rate in effect at the time for this population was substantially higher at $1085, generating an annual "risk pool" of $9.1 million. Care provided in the two facilities varied greatly. Hospitalization rates, clinic-based primary care and specialist visits, and therapy sessions were greater in facility one. Overall expenditures were lower for residents at facility two, where the majority of care was provided by trained geriatricians in collaboration with physician extenders and without sophisticated clinical pathways and utilization controls. CONCLUSIONS: Our data support other studies that suggest that teams of geriatricians and physician extenders can reduce hospitalization rates and overall expenditures. Capitated rates for the frail, geriatric population warrant careful study. These rates must balance fiscal responsibility with the need for adequate, risk-adjusted payments that create incentives for providers to produce high quality as well as cost-effective care. PMID- 11037024 TI - Medicare prescription drug statement of principles. American Geriatrics Society Clinical Practice Committee. PMID- 11037025 TI - Care management position statement. American Geriatrics Society. PMID- 11037026 TI - Protecting long-term care patients from antibiotic resistant infections: ethics, cost-effectiveness, and reimbursement issues. PMID- 11037027 TI - Improving advance care planning: lessons from POLST. Physician Orders for Life Sustaining Treatment. PMID- 11037028 TI - Neuroimaging in the dementia assessment: is it necessary? PMID- 11037029 TI - Treatment guideline for nursing home-acquired pneumonia. PMID- 11037030 TI - Vascular dementia presenting as Lewy-body dementia. PMID- 11037031 TI - Education and care at the end of life. PMID- 11037032 TI - Carotid wall shear stress and aging. PMID- 11037033 TI - Efficacy of iloprost in nonexudative, age-related macular degeneration. PMID- 11037034 TI - Interfacing mind and brain: a neurocognitive model of recognition memory. AB - A variety of processes contribute to successful recognition memory, some of which can be associated with spatiotemporally distinct event-related potential old/new effects. An early frontal and a subsequent parietal old/new effect are correlated with the familiarity and recollection subcomponents of recognition memory, respectively, whereas a late, postretrieval old/new effect seems to reflect an ensemble of evaluation processes that are set by the task context in which retrieval occurs. Both the early frontal and the parietal old/new effects are differentially modulated by the informational content (e.g., object forms and spatial locations) of recognition and seem to rely on brain systems damaged in amnesia. The late frontal effect appears to reflect prefrontal cortex activation. A neurophysiologically based model of recognition memory retrieval is presented and it is shown that coupling recognition memory subprocesses with distinct old/new effects allow examination of the time course of the processes that contribute to correct and to illusory memories. In conjunction with event-related functional magnetic resonance imaging activation patterns the brain systems recruited by various aspects of episodic memory retrieval can be identified. PMID- 11037035 TI - Effects of nicotine and caffeine, separately and in combination, on EEG topography, mood, heart rate, cortisol, and vigilance. AB - Effects of nicotine and caffeine, separately and in combination, were assessed in 12 male habitual smokers in a repeated-measures design. Caffeine (0-mg vs. two 150-mg doses administered in a decaffeinated/sugar-free cola drink post-baseline and 90 min later) was crossed with nicotine (ad libitum own dosing vs. 1.0-mg machine-delivered dose vs. 0.05-mg machine-delivered dose). Participants smoked a total of five cigarettes at 30-min intervals over a 2-hr period. Caffeine and nicotine had large effect sizes on electroencephalogram (EEG) power; however, these effects were modulated by the eyes open versus closed condition, the other drug, and electrode site. EEG effects of open versus closed eyes tended to be of the same size and direction as those of nicotine and caffeine. However, whereas nicotine increased EEG power in some higher frequency bands in some conditions, caffeine decreased EEG power across almost all conditions. Serum cortisol concentration, vigor, and pleasantness were increased by nicotine, but not by caffeine. Level of depressive mood depended on an interaction of caffeine and nicotine. Vigilance performance was enhanced significantly by caffeine and was increased almost significantly by nicotine. The findings were interpreted in terms of common and differential mechanisms of the two drugs. PMID- 11037036 TI - Short duration power changes in the EEG during recognition memory for words and faces. AB - Although memory has been widely studied using event-related potentials, memory related changes in the electroencephalogram (EEG) have been relatively neglected. The aim of this study was to determine whether evidence could be found for memory related changes in the EEG. EEG was recorded from a sample of healthy volunteers while they performed word and face recognition memory tasks. Data were analyzed using the method of event-related desynchronization. In the theta frequency range there was a short-duration increase in power that occurred in the first 250 ms that was maximal at temporal sites (T5/T6). For words, but not faces, there was a repetition effect in theta such that new words elicited greater synchronization than old words at the midline frontal electrode (Fz). In the alpha frequency range there was a lateralized repetition effect, which occurred from 750 ms. In upper alpha this effect was lateralized in the expected way with greater desynchronization at temporo-parietal sites on the left for words and on the right for faces. For lower alpha, the lateralization was reversed. The meanings of these findings are interpreted in the light of existing models of recognition memory. PMID- 11037037 TI - Selective inhibition is indexed by heart rate slowing. AB - This study examined the hypothesis that global and selective inhibition are mediated by distinct mechanisms: respectively, a peripheral mechanism, indexed by heart rate slowing, and a central mechanism, indexed by cortical but not autonomic measures. Three varieties of a Go-NoGo task were presented in which the Go signal required an index finger response rapidly followed by a middle finger response. The NoGo signal required the inhibition of (a) both responses (global inhibition), (b) the middle finger response (simple selective inhibition), or (c) the index finger response of one hand and the middle finger response of the other hand (complex selective inhibition). As anticipated, global inhibition was indexed by heart rate slowing. Most importantly, heart rate slowing was also elicited by selective inhibition and was more pronounced for complex than simple selective inhibition. These findings suggest that global and selective inhibition are mediated by one rather than two mechanisms and that heart rate is sensitive to the demands placed on this inhibition mechanism. PMID- 11037038 TI - Mental fatigue and task control: planning and preparation. AB - The effects of mental fatigue on planning and preparation for future actions were examined, using a task switching paradigm. Fatigue was induced by "time on task," with subjects performing a switch task continuously for 2 hr. Subjects had to alternate between tasks on every second trial, so that a new task set was required on every second trial. Manipulations of response-stimulus intervals (RSIs) were used to examine whether subjects prepared themselves for the task change. Behavioral measurements, event-related potentials (ERPs), and mood questionnaires were used to assess the effects of mental fatigue. Reaction times (RTs) were faster on trials in which no change in task set was required in comparison with switch trials, requiring a new task set. Long RSIs were used efficiently to prepare for the processing of subsequent stimuli. With increasing mental fatigue, preparation processes seemed to become less adequate and the number of errors increased. A clear poststimulus parietal negativity was observed on repetition trials, which reduced with time on task. This attention-related component was less pronounced in switch trials; instead, ERPs elicited in switch trials showed a clear frontal negativity. This negativity was also diminished by time on task. ERP differences between repetition and switch trials became smaller with increasing time on task. PMID- 11037039 TI - Developmental change in auditory selective attention as reflected by phasic heart rate changes. AB - Heart rate was recorded from five different groups of children (ages 7, 10, 12, 14, and 20 years) while they were performing an auditory selective attention task. The participants were instructed to count rare tone pips embedded in a series of standard tone pips presented at one (attended) ear while ignoring rare and standard stimuli presented at the other (unattended) ear. A pattern of anticipatory heart rate deceleration followed by acceleration was associated with rare tone pips at the attended ear but not with rare tone pips that should be ignored. The absence of differential sensitivity of heart rate responses to rare tone pips presented at the unattended ear was observed for all age groups. These findings were interpreted to suggest that the ability to ignore irrelevant target stimuli has reached mature levels during middle childhood. The depth of anticipatory deceleration increased until age 14, suggesting that the ability to maintain attentional set continues to develop beyond childhood. PMID- 11037040 TI - Secretory immunoglobulin A and cardiovascular reactions to mental arithmetic, cold pressor, and exercise: effects of alpha-adrenergic blockade. AB - The mechanism underlying acute changes in secretory immunoglobulin A (sIgA) remains to be determined. In this experiment, sIgA and cardiovascular activity were monitored at rest and while participants performed a mental arithmetic task, cold pressor, and submaximal cycle exercise following placebo or 1 mg of the alpha-adrenergic blocker, doxazosin. Under placebo, the tasks produced patterns of cardiovascular activity indicative of combined alpha- and beta-adrenergic, alpha-adrenergic, and beta-adrenergic activation, respectively. Doxazosin was associated with reduced blood pressure during cold pressor, but not during arithmetic or exercise. Mental arithmetic elicited increases in sIgA concentration and exercise produced increases in both sIgA concentration and secretion rate; these changes were unaffected by alpha blockade. In contrast, the cold pressor was associated with decreases in both sIgA concentration and secretion rate, which were blocked by doxazosin. These data suggest that acute decreases, but not increases, in sIgA are mediated by alpha-adrenergic mechanisms. PMID- 11037041 TI - Relationship between P300 amplitude and subsequent recall for distinctive events: dependence on type of distinctiveness attribute. AB - Distinctive words elicit the P300 component of the event-related brain potential, and are also likely to be recalled. Previous studies have shown that the larger the P300 elicited by distinctive words, the more likely it is that those words will be recalled. The present study addressed whether this relationship is affected by the manner in which distinctiveness is induced. Distinctiveness was manipulated either by varying the size of the characters in which a word was displayed, or by surrounding the word with a frame at close or far distance. All distinctiveness attributes resulted in improved recall performance. The words whose size was distinctive elicited a large P300, and P300 amplitude was larger for subsequently recalled words. The frame attributes elicited a small P300, and the amplitude of these P300s was not correlated with subsequent recall performance. Instead, a frontal slow wave was correlated with subsequent recall performance in the far frame group. It is concluded that the relationship between P300 amplitude and subsequent recall depends on the type of distinctiveness attribute, and should therefore not be ascribed to a generalized effect of distinctiveness on memory encoding processes. PMID- 11037042 TI - Location selection in the visual domain. AB - According to A.H.C. Van der Heijden (1992), attentional selection of visual stimuli can be considered as location selection. Depending on the type of task, location selection can be considered to be automatic (e.g., in case of abrupt onsets), directly controlled (e.g., in case of symbolic precues), or indirectly controlled (e.g., in case of visual search). In tasks with symbolic precues a contralateral enhancement of the P1 event-related potential (ERP) component has been found, whereas in visual search tasks a contralateral enhancement of the N2 component has been found. We hypothesized that both lateralized effects reflect the influence of location selection, which differs only in its moment of activation. Linear arrays (a target among five distractors, presented at two eccentricities) requiring left or right responses, were preceded by cues indicating either the precise target location or the side of the target, or by bilateral cues that indicated all array positions. Array-evoked ERPs, corrected for cue-evoked and interactive effects, showed not only lateralized but also target location-specific effects for both components, which supports the view that, depending on the type of task, either the P1 or the N2 component reflect the influence of location selection. PMID- 11037043 TI - Event-related brain potential and heart rate manifestations of visual selective attention. AB - Twenty-eight volunteers were instructed to attend stimuli presented at one side of the computer screen and to ignore stimuli presented at the other side. Both attended and unattended stimulus series consisted of targets (25%) and nontargets (75%) defined on the basis of stimulus shape. Attended targets required a binary choice based on stimulus color. Selective attention led to the expected increase in both midlatency (N2b) and late (P3) brain potential components. Furthermore, selective attention led to increased anticipatory cardiac slowing preceding the target stimulus and to increased primary bradycardia. Correlational analyses revealed a positive relation between the effects of selective attention on N2b amplitude and primary bradycardia suggestive of cortical involvement in the chronotropic control of heart rate. PMID- 11037044 TI - Valence-dependent modulation of psychophysiological measures: is there consistency across repeated testing? AB - The present study used the picture perception paradigm to examine the extent to which three well-documented psychophysiological measures demonstrate consistency across time in response to emotional stimuli. The three measures were the eye blink startle response and the activation in two facial muscle regions (zygomatic and corrugator). Twenty-seven young women were assessed on two occasions, 2 weeks apart. Whereas activation in the corrugator and zygomatic muscle regions demonstrated the predicted patterns at both assessments (with some attenuation in the zygomatic muscle regions), the startle response had limited consistency across the two assessments. The startle response revealed the predicted linear pattern of valence modulation during the first assessment. During the second assessment, startle magnitude response was a quadratic function of valence ratings and a linear function of arousal ratings. The unexpected pattern of startle response during the second session appeared to be related to the content of the pleasant slides, with action slides generating quadratic valence modulation and erotic slides continuing to exhibit the expected linear valence modulation. PMID- 11037045 TI - Facial reactions to happy and angry facial expressions: evidence for right hemisphere dominance. AB - Previous research on asymmetric effects of emotional expression and brain hemispheric asymmetry has supported opposing theories of hemispheric dominance in the control of emotional reactions. In the present study, 32 subjects were exposed to pictures of happy and angry facial stimuli while facial electromyographic (EMG) activity from the zygomatic major and the corrugator supercilii muscle regions was detected from the left and right sides of the face. The subjects reacted spontaneously and rapidly with larger zygomatic EMG activity to happy facial stimuli and larger corrugator EMG activity to angry stimuli. These distinct reactions were significantly larger on the left side of the face. It is concluded that the present results support the hypothesis that the right brain hemisphere is predominantly involved in the control of spontaneously evoked emotional reactions. PMID- 11037046 TI - An event-related brain potential study of cross-modal links in spatial attention between vision and touch. AB - Event-related potential (ERP) evidence for the existence of cross-modal links in endogenous spatial attention between vision and touch was obtained in an experiment where participants had to detect tactile or visual targets on the attended side and to ignore the irrelevant modality and stimuli on the unattended side. For visual ERPs, attentional modulations of occipital P1 and N1 components were present when attention was directed both within vision and within touch, indicating that links in spatial attention from touch to vision can affect early stages of visual processing. For somatosensory ERPs, attentional negativities starting around 140 ms poststimulus were present at midline and lateral central electrodes when touch was relevant. No attentional somatosensory ERP modulations were present when vision was relevant and tactile stimuli could be entirely ignored. However, in another task condition where responses were also required to infrequent tactile targets regardless of their location, visual-spatial attention modulated somatosensory ERPs. Unlike vision, touch apparently can be decoupled from attentional orienting within another modality unless it is potentially relevant. PMID- 11037047 TI - Image motion and context: a between- and within-subjects comparison. AB - In two previous experiments, we studied how stimulus motion affects both the self report of emotion experience and the physiological sequelae of emotion. In both studies, image motion intensified emotional responding, and the effect of motion was relatively specific to the arousal dimension of the emotion; there was little evidence that image motion altered the valence of the image. Moving images also appeared to sustain the attention of the participants for a longer period of time than did the still images. In these two experiments, however, image motion was manipulated within participants. In the present experiment, we used a between subjects manipulation of image motion and found a nearly identical pattern of results. These data indicate that motion inherently increments the arousal value of an image and that this increment is not dependent on the context in which motion is introduced. PMID- 11037048 TI - Focused attention in anhedonia: a P3 study. AB - Attentional and information processing impairments have been evidenced in nonclinical anhedonic subjects. However, the extent of attentional deficit has not been determined. We studied focused attention, the ability to reject irrelevant or distracting messages, in anhedonic nonclinical subjects. The event related potentials and behavioral performances of anhedonic subjects were compared with those of control subjects during the Eriksen focused attention task (C.W. Eriksen & B.A. Eriksen, 1974); the task combined one compatible and one incompatible condition, the latter causing an interference. Anhedonic subjects exhibited a smaller P300 and slower reaction times than control subjects. Varying task conditions had different effects on anhedonic subjects and controls, suggesting that anhedonic subjects may have developed a conservative response strategy. In view of previous works, these results suggest that attentional impairment is not ascribed to specific processes, but may involve a more global deficit, that is, a resource allocation deficit. PMID- 11037049 TI - Current perspectives of bispecific antibody-based immunotherapy. AB - The field of bispecific antibodies is an evolving field of research that has increasing clinical appeal. The fusion of two antibodies or antibody fragments introduced a new way to override natural specificity of T cell and induce effector responses against tumor targets in MHC-unrestricted manner. Initial experiences with bispecific antibodies demonstrate both the promise for and limitations of this anti-cancer strategy. Significant body of work has shown that bispecific antibodies have potential to induce T cell mediated anti-tumor responses in pre-clinical models. However, immunotherapy with bispecific antibodies in humans has yet to prove its value in clinical settings. In addition, the production of high-quality bispecific antibodies for clinical applications, the optimal size and avidity of bispecific antibodies, and in vivo T cell pre-activation remain critical issues. In this review, we summarize recent progress in bispecific antibody-based immunotherapy and address essential aspects of this anti-cancer strategy. PMID- 11037050 TI - Molecular diagnostics in the treatment of childhood acute lymphoblastic leukemia. AB - Somatically acquired genetic alterations play an important role in the pathogenesis of acute lymphoblastic leukemia. The molecular analysis of these alterations has increased our understanding of the mechanisms of leukemogenesis. In addition, this information has led to improvements in our abilities to predict treatment response and to deliver the optimal intensity of treatment to individual patients. For example, the prognosis for patients with acute lymphoblastic leukemia whose leukemic cells express the TEL-AML1 fusion is favorable when they are treated on modem chemotherapy protocols, whereas patients whose leukemic lymphoblasts contain the MLL-AF4 or the BCR-ABL fusion sometimes require allogeneic hematopoietic stem cell transplantation for cure. Molecular techniques are also used to detect minimal residual disease and genetic polymorphisms that are important in optimizing drug therapy. PMID- 11037051 TI - P202: an interferon-inducible negative regulator of cell growth. PMID- 11037052 TI - The distribution of the major peripheral blood T, B and NK cell subsets does not predict the clinical outcome of Graves' disease patients after methimazole therapy. AB - Biological markers capable of predicting the clinical outcome of antithyroid drug therapy could be clinically useful in selecting the modality of treatment for Graves' disease, but at present they are unavailable. In the present study we prospectively explore the value of 22 different peripheral blood T, B and NK lymphocyte subsets to predict remission and relapse in a group of 42 Graves' disease patients. Eighteen patients were studied at diagnosis, before treatment, and 24 during antithyroid drug therapy. All cases were followed-up for at least one year after finishing an 18 month cycle of methimazole therapy. The combination of flow cytometry and 3- color immunofluorescence did not reveal significant differences in the distribution of the major peripheral blood T, B and NK cell subsets between the relapsed patients and those in remission, both in the groups studied at diagnosis and in those analyzed during the cycle of antithyroid drug therapy. In our search for a prognostic marker for relapse prediction we found that some lymphoid subpopulations such as total B cells, total NK and NK CD8+ cells showed high sensitivity (88-100%). In turn, other subsets such as TCD8+, total T and B cells expressing the CD25 antigen displayed high specificity (77-88%). PMID- 11037053 TI - Abrogation of surgery-induced decline in circulating dendritic cells by subcutaneous preoperative administration of IL-2 in operable cancer patients. AB - Surgery-induced immunosuppression is characterized by a decline in lymphocyte count, particularly T lymphocyte number. In addition, preliminary studies have shown that the postoperative period is also characterized by a decline in the number of circulating dendritic cells (DC), whose fundamental anticancer role has been recently demonstrated. Previous studies had already shown that the preoperative injection of IL-2 may completely abrogate surgery-induced lymphocytopenia, whereas its eventual influence on DC system during the perioperative period is still unknown. The present study was performed to evaluate the influence of IL-2 preoperative immunotherapy on the perioperative changes in circulating DC number in patients affected by colorectal cancer. The study included 14 consecutive patients, who were randomized to be treated with or without IL-2 presurgical immunotherapy (12 million IU/day for 3 days subcutaneously). Circulating immature and mature cells were evaluated before surgery and at days 3 and 7 of the postoperative period. The detection was made by FACS using monoclonal antibodies against CD123 and CD11c to recognize immature and mature DC, respectively. Surgery induced a significant decline in the mean number of both immature and mature DC. The pre-surgical administration of IL-2 completely abrogated surgery-induced decline in immature DC cell amount. Moreover, mature DC mean number was diminished only at day 3 of the postoperative period, since the value observed at day 7 was not significantly lower than that found before surgery. This preliminary study shows that surgery-induced immunosuppression is characterized also by a significant decline in the mean number of both immature and mature DC. Moreover, this study would suggest that the preoperative immunotherapy with IL-2 may counteract surgery-induced failure of DC system. Because of the fundamental antitumor role of DC, this evidence could have a prognostic impact on the clinical course of the neoplastic disease. PMID- 11037054 TI - Dynamics of diurnal changes in serum concentration of TNF-alpha soluble receptors in gastrointestinal cancer patients. AB - The objective of the study was to investigate the dynamic changes in soluble TNFalpha p-55 and p-75 receptors in serum of advanced cancer patients during 24 hours. The group examined consisted of 42 patients suffering from advanced gastrointestinal neoplasms (colorectal, gastric and pancreatic cancer). Serum levels of the cytokine and both receptors in cancer patients were measured using ELISA type kits 6 times a day (8.00 a.m., 2.00 p.m., 6.00 p.m., 10.00 p.m., 2.00 a.m. and again 8.00 a.m.) as well as in healthy controls. The levels of TNFalpha and its soluble receptors were substantially increased in the examined group and displayed statistically significant circadian fluctuations. The presence of circadian rhythm of the cytokine was proved (acrophase - 00.36 a.m.), however no diurnal rhythm of soluble TNF receptors was observed. The concentration of p-55 receptor was distinctly lower then p-75. The peak p-55 value appeared at 10.00 p.m. while the p-75 reached its minimum level at the same time. Although there was no statistical correlation between the receptor concentrations the shapes of both curves remained inversely proportional. The present results may suggest the presence of complex self-regulation mechanisms in advanced gastrointestinal cancer patients. PMID- 11037055 TI - Inhibition of HIV-1 transcription by cyclopentenone prostaglandin A1 in Jurkat T lymphocytes. AB - Cyclopentenone prostaglandins inhibit virus replication in several DNA and RNA virus models. In this report we investigated the effect of prostaglandin A1 (PGA1) on HIV-1 transcription in human CD4+ Jurkat T lymphocyte cells. A dramatic reduction of HIV-1 RNA levels was detected up to seven days post infection in both unstimulated and phorbol 12-mystrate 13-acetate (PMA)-stimulated cells treated with PGA1- PGA1 treatment of cells was also effective in inhibiting the transcription of a chloramphenicol acetyltransferase (CAT) reporter gene, under the control of HIV-1 LTR, in Jurkat-Tat cells. We also show that PGA1 induced the synthesis of 70-kDa heat-shock protein (HSP70) in this cell system and the induction correlated with the drug-antiviral activity. PGA1 was also found to induce the loss of the tumor suppressor p53 protein, in the "proliferative" conformation, in a time correlation with the induction of the HSP70 As the "proliferative" p53 has been involved in the positive trans-activation of the HIV 1 LTR its depletion could contribute to the inhibitory mechanisms of PGA1 on virus transcription. PMID- 11037056 TI - Markers of cell lineage, differentiation and activation. AB - The most widespread use of CD markers is in the determination of cell lineage and sublineage. For example, T cells are identified by the expression of CD3 (reviewed in this issue of CD corner). A mature T cell may belong to the T4 subset, in which case it will express CD4. Similarly, there are markers for other cell populations and sub-populations. Within the lineages, it is helpful to distinguish cells at different stages of differentiation and activation. Differentiation status is particularly useful in the diagnostic analysis of the lymphoid and myeloid malignancies, and in research on the haemopoietic system. Examples include markers for naive or antigen-experienced cells (especially the CD45 isoforms) and molecules such as CALLA (CD9) found on B-lineage precursors, including B lineage acute lymphoblastic leukaemia. Activation status is especially interesting in studies of cell function. Activation markers include growth factor receptors such as CD25 (a component of the receptor for IL-2), and molecules who's cellular function is not fully understood, such as CD69 and CD98. These markers have revolutionised aspects of pathology and research, and the ease with which some cell populations can be identified has lead to some unrealised, and perhaps unrealistic, expectations. We expect to be able to identify T helper type 1 (TH1) and T helper type 2 (TH2) cells on the basis of a simple surface marker; we are frustrated by the lack of a single marker for all dendritic cells or for all NK cells; we are confused by the un-coordinated expression of different activation markers; we tend to over-interpret phenotype in some situations. To find solutions to these problems it is helpful to examine why the successful lineage markers work so well, and to reconsider our expectations. Lineage markers are the main focus in this commentary; the question of activation markers and markers of differentiation state will be considered in a separate paper. PMID- 11037057 TI - CD148. PMID- 11037058 TI - CD2. PMID- 11037059 TI - CD3 complex. PMID- 11037060 TI - CD5. Other names: T1, Leu-1, Tp67 (human), and Ly-1 (mouse). PMID- 11037061 TI - CD6. PMID- 11037062 TI - CD27. Other names: Tp55. PMID- 11037063 TI - Everything is connected. PMID- 11037064 TI - Long-term career attainments of deaf and hard of hearing college graduates: results from a 15-year follow-up survey. AB - This article reports on the results of a national longitudinal survey of 240 college graduates with hearing loss. Results confirm that economic benefits resulted from these alumni's postsecondary training. Most respondents were relatively successfully employed and satisfied with life. Over time, increasing numbers had completed higher degrees and secured white-collar positions. Between 1988 and 1998, men in the study sample made more consistent earnings gains than their female counterparts. Larger proportions of deaf alumni had earned advanced degrees and secured white-collar jobs than hard of hearing alumni. Deaf alumni also earned more. Results also showed that recipients of associate's degrees earned more than recipients of bachelor's degrees. Implications of the findings for secondary educators, vocational rehabilitation counselors, and postsecondary service providers are discussed. Recommendations are made on how to improve career decision making by deaf and hard of hearing adolescents, enrich the career potential of deaf and hard of hearing women, and increase the productivity of workers with hearing loss. PMID- 11037065 TI - The sign language skills classroom observation: a process for describing sign language proficiency in classroom settings. AB - In collaboration with teachers and students at the National Technical Institute for the Deaf (NTID), the Sign Language Skills Classroom Observation (SLSCO) was designed to provide feedback to teachers on their sign language communication skills in the classroom. In the present article, the impetus and rationale for development of the SLSCO is discussed. Previous studies related to classroom signing and observation methodology are reviewed. The procedure for developing the SLSCO is then described. This procedure included (a) interviews with faculty and students at NTID, (b) identification of linguistic features of sign language important for conveying content to deaf students, (c) development of forms for recording observations of classroom signing, (d) analysis of use of the forms, (e) development of a protocol for conducting the SLSCO, and (f) piloting of the SLSCO in classrooms. The results of use of the SLSCO with NTID faculty during a trial year are summarized. PMID- 11037066 TI - College teachers' perceptions of English language characteristics that identify English language learning disabled deaf students. AB - Deaf individuals typically experience English language difficulties at all levels of linguistic knowledge. Hearing individuals with English language learning disabilities (LD) can exhibit the same kinds of English language difficulties as deaf individuals. Although the existence of deaf individuals who also have LD has long been recognized, no definite criteria for identifying them exist, partly because of the confounding effects of deafness and LD on English language development. Despite the confound, previous surveys suggest that teachers believe atypical English-language behavior is a potential diagnostic marker for LD in deaf individuals. In the present study, a survey solicited the intuitions of experienced teachers and tutors of English to deaf college students regarding the degree of difficulty deaf students with and without LD might be expected to have in dealing with 30 specific English language phenomena. Spelling knowledge and a variety of English discourse, lexical syntactic, and morphological phenomena emerged as candidates for further study as potential markers of LD in the deaf population. PMID- 11037067 TI - Beyond parent education: the impact of extended family dynamics in deaf education. AB - The impact of childhood deafness on family dynamics, with particular attention given to the influence and role of grandparents, is discussed. Models for the successful involvement of parents in the education of deaf children are already in place and can be applied to extended family, especially grandparents, with promising results. An informal questionnaire focusing on grandparents' support, involving 10 hearing parents of deaf infants and preschool children, is reviewed. PMID- 11037068 TI - "A good future for deaf children": a five-year sign language intervention project. AB - Deaf preschoolers and hearing family members learned sign language in a 5-year intervention project. Once weekly, each child met with a teacher who was deaf. Parents, siblings, and other relatives met about once monthly to study sign language, and all families in the project signed together about twice yearly. The present study addressed four questions asked of parents about the project: (a) How did the children learn to sign? (b) Did both the parents and the children benefit from the project? (c) What was the position of sign language in the family? (d) Did the project have some impact on the family's social network? The families indicated satisfaction with the project; they learned to sign and their social networks expanded. Parents favored bilingual education: Sign language was the main language but learning Finnish was also important. Learning sign language was not easy, especially for the fathers. The families that were most actively involved in the lessons learned the most. PMID- 11037069 TI - Factors contributing to parents' selection of a communication mode to use with their deaf children. AB - This study examines factors contributing to parents' selection of a communication mode to use with their children with hearing loss. More than 90% of children with prelingual hearing loss have normally hearing parents. Communication difficulties are among the obstacles facing these parents in connection with these children's development. Controversy over manual and aural/oral methods of communication creates further complications. Case studies of two families with deaf children were conducted to identify factors that could influence parents' selection of a communication method. Semistructured questionnaires and unstructured interviews were used in data collection. A qualitative approach was used in data analysis. Based on the results, the factors influencing parental choice were grouped under four themes: (a) the influence of information provided to parents, (b) parents' perceptions of assistive technology, (c) attitudes of service professionals and educational authorities, and (d) quality and availability of support services. Implications of these themes for service provision are discussed. PMID- 11037070 TI - Treating fibromyalgia: science vs. art. PMID- 11037071 TI - Generalized anxiety disorder in family practice patients. PMID- 11037072 TI - Foreign-body aspiration, asthma and chronic bronchitis. PMID- 11037073 TI - Withholding and withdrawing life-sustaining treatment. AB - Withholding or withdrawing life-sustaining therapies is ethical and medically appropriate in some circumstances. This article summarizes the American Medical Association's Education for Physicians on End-of-life Care (EPEC) curriculum module on withholding or withdrawing therapy. Before reviewing specific treatment preferences, it is useful to ask patients about their understanding of the illness and to discuss their values and general goals of care. Family physicians should feel free to provide specific advice to patients and families struggling with these decisions. Patients with decision-making capacity can opt to forego any medical intervention, including artificial nutrition/hydration and cardiopulmonary resuscitation. PMID- 11037074 TI - Anemia in the elderly. AB - Anemia should not be accepted as an inevitable consequence of aging. A cause is found in approximately 80 percent of elderly patients. The most common causes of anemia in the elderly are chronic disease and iron deficiency. Vitamin B12 deficiency, folate deficiency, gastrointestinal bleeding and myelodysplastic syndrome are among other causes of anemia in the elderly. Serum ferritin is the most useful test to differentiate iron deficiency anemia from anemia of chronic disease. Not all cases of vitamin B12 deficiency can be identified by low serum levels. The serum methylmalonic acid level may be useful for diagnosis of vitamin B12 deficiency. Vitamin B12 deficiency is effectively treated with oral vitamin B12 supplementation. Folate deficiency is treated with 1 mg of folic acid daily. PMID- 11037075 TI - Treating fibromyalgia. AB - Fibromyalgia is an extremely common chronic condition that can be challenging to manage. Although the etiology remains unclear, characteristic alterations in the pattern of sleep and changes in neuroendocrine transmitters such as serotonin, substance P, growth hormone and cortisol suggest that dysregulation of the autonomic and neuroendocrine system appears to be the basis of the syndrome. The diagnosis is clinical and is characterized by widespread pain, tender points and, commonly, comorbid conditions such as chronic fatigue, insomnia and depression. Treatment is largely empiric, although experience and small clinical studies have proved the efficacy of low-dose antidepressant therapy and exercise. Other less well-studied measures, such as acupuncture, also appear to be helpful. Management relies heavily on the physician's supportive counseling skills and willingness to try novel strategies in refractory cases. PMID- 11037076 TI - Generalized anxiety disorder. AB - Patients with generalized anxiety disorder experience worry or anxiety and a number of physical and psychologic symptoms. The disorder is frequently difficult to diagnose because of the variety of presentations and the common occurrence of comorbid medical or psychiatric conditions. The lifetime prevalence is approximately 4 to 6 percent in the general population and is more common in women than in men. It is often chronic, and patients with this disorder are more likely to be seen by family physicians than by psychiatrists. Treatment consists of pharmacotherapy and various forms of psychotherapy. The benzodiazepines are used for short-term treatment, but because of the frequently chronic nature of generalized anxiety disorder, they may need to be continued for months to years. Buspirone and antidepressants are also used for the pharmacologic management of patients with generalized anxiety disorder. Patients must receive an appropriate pharmacologic trial with dosage titrated to optimal levels as judged by the control of symptoms and the tolerance of side effects. Psychiatric consultation should be considered for patients who do not respond to an appropriate trial of pharmacotherapy. PMID- 11037078 TI - Photo quiz. Facial masses. PMID- 11037077 TI - A family physician's guide to monitoring methotrexate. AB - Methotrexate has a long history of use in the treatment of various immunologic diseases, including rheumatoid arthritis and psoriasis. Although the drug is usually prescribed by a subspecialist, a family physician may assume responsibility for monitoring methotrexate therapy. Major toxic effects, such as hepatic, pulmonary, renal and bone marrow abnormalities, require careful monitoring. Minor toxic effects, such as stomatitis, malaise, nausea, diarrhea, headaches and mild alopecia, are common but respond to folate supplementation. Methotrexate is administered once weekly as a single dose or in divided doses given over a 24-hour period. To reduce the incidence of major toxic effects, methotrexate should never be given in daily doses. Relative contraindications include renal dysfunction, liver disease, active infectious disease and excessive alcohol consumption. Both women and men of reproductive age should use birth control during methotrexate therapy. Potential drug interactions include salicylates and nonsteroidal anti-inflammatory drugs, which are both commonly used in patients with rheumatoid arthritis or psoriasis. A premethotrexate evaluation is important to ensure proper patient selection for this effective but potentially toxic drug. PMID- 11037079 TI - Headache Consortium releases guidelines for use of CT or MRI in migraine work-up. PMID- 11037080 TI - A consultant takes over. PMID- 11037081 TI - Patient education. American Academy of Family Physicians. PMID- 11037082 TI - The need for treatment evidence for common mental disorders in developing countries. PMID- 11037083 TI - Randomized controlled trial of cognitive behaviour therapy for repeated consultations for medically unexplained complaints: a feasibility study in Sri Lanka. AB - BACKGROUND: Research on the management and the outcome of treatment of medically unexplained symptoms is very limited. Development of simple but effective techniques for treatment and demonstration of their effectiveness when applied in primary health care are needed. METHODS: A randomized controlled trial was carried out with follow-up assessments at 3 months after baseline assessments using the Short Explanatory Model Interview (SEMI), General Health Questionnaire (GHQ-30), Bradford Somatic Inventory (BSI) and patient satisfaction on a visual analogue scale. The study was carried out in a general out-patient clinic in Sri Lanka. The intervention group received six, 30 min sessions based on the principles of cognitive behavioural therapy over a period of 3 months. The control group received standard clinical care. RESULTS: Eighty patients out of the 110 patients referred, were eligible. Sixty-eight were randomly allocated equally to the control and treatment groups. All 34 in the treatment group accepted the treatment offer and 22 completed between three and six sessions. At 3 months, 24 in the treatment and 21 in the control group completed follow-up assessments. Intention-to-treat analysis revealed significant differences in mean scores of outcome measures (adjusted for baseline scores) between control and intervention groups respectively--complaints 6.1 and 3.8 (P = 0.001), GHQ 10.4 and 6.3 (P = 0.04), BSI score 15.6 and 132 (P < 0-01), visits 7.9 and 3.1 (P = 0.004). CONCLUSIONS: Intervention based on cognitive behavioural therapy is feasible and acceptable to patients with medically unexplained symptoms from a general out-patients clinic in Sri Lanka. It had a significant effective in reducing symptoms, visits and distress, and in increasing patient satisfaction. PMID- 11037084 TI - Familial aggregation for conduct disorder symptomatology: the role of genes, marital discord and family adaptability. AB - BACKGROUND: There is extensive evidence of statistical associations between family discord/ maladaptation and antisocial behaviour in the children, but questions remain on the extent to which the psychopathological risks are genetically or environmentally mediated. METHODS: Twin pairs (N = 1,350), aged 8 to 16 years, in the general population-based Virginia Twin Study of Adolescent Behavioral Development were assessed using the Child and Adolescent Psychiatric Assessment interview administered separately to both twins and both parents. Structured interviews for parental lifetime psychiatric disorders were also administered to the mothers and fathers. Maternal reports on Olsson's Family Adaptability and Cohesiveness questionnaire and the Dyadic Adjustment Scale were used as indices of the family environment. A path analytical model based on an extended twin-family design was used to test hypotheses about parent offspring similarity for conduct disorder symptomatology. RESULTS: Family discord and maladaptation, which intercorrelated at 0.63, were associated with a roughly two fold increase in risk for conduct disorder symptomatology. When parental conduct disorder was included in the model the environmental mediation effect for family maladaptation remained, but that for family discord was lost. CONCLUSION: It is concluded that there is true environmental mediation from family maladaptation, operating as a shared effect, which accounts for 3.5 % of the phenotypic variance. The assumptions underlying this genetic research strategy are made explicit, together with its strengths and limitations. PMID- 11037085 TI - Cohort differences in genetic and environmental influences on retrospective reports of conduct disorder among adult male twins. AB - BACKGROUND: Rates of child and adolescent conduct disorder (CD) have increased steadily over the past several decades. What is not known is whether the underlying genetic and environmental influences on individual differences in CD have also changed. METHODS: Retrospective reports of antisocial behaviour prior to age 18 were obtained from a population-based sample of 2769 adult males from male-male twin pairs born between 1940 and 1974. Using a summary score of number of CD symptoms, structural equation modelling was used to investigate whether mean level and variation in CD increased with more recent cohorts, and whether any increase in variance could be explained by familial or non-familial factors. RESULTS: Both mean level CD symptoms and variation were increased in more recent cohorts. Model fitting indicated that the primary increase in variance was due to familial factors, most notably, an increase in the shared environmental influences on CD, from 0.01 (95 % CI = 0.00; 0.27) to 0.30 (95 % Cl = 0.01; 0 44). Heritability estimates remained largely unchanged, although an increase in genetic factors could not be ruled out. CONCLUSIONS: Secular changes in sociodemographic factors responsible for increasing rates of CD may also account for the greater magnitude of shared environmental influences on variation in CD found among more recent cohorts. PMID- 11037086 TI - Birth order and ratio of brothers to sisters in transsexuals. AB - BACKGROUND: As previous studies with homosexual males have revealed a later birth order, more older brothers and more brothers than sisters, this research was extended to a large series of transsexual males and females, some of whom are homosexual. METHODS: The male sample comprised 442 male-to-female transsexuals, subdivided by sexual partner preference: 106 homosexual, 135 heterosexual, 155 bisexual and 46 asexual. One hundred female-to-male transsexuals were also studied: 75 homosexual, 16 bisexual, seven heterosexual and five asexual. Birth order was computed by both Slater's Index and Berglin's Index. RESULTS: Homosexual male-to-female transsexuals have a later than expected birth order and more older brothers than other subgroups of male-to-female transsexuals. Each older brother increases the odds that a male transsexual is homosexual by 40 %. CONCLUSIONS: Hypotheses explaining the extension of prior findings to this large sample of transsexual males include a progressive maternal immunization to the male foetus either through the H-Y antigen or protein-bound testosterone or alterations in foetal androgen levels in successive pregnancies, all modifying male psychosexual development. Data on the sexual orientation of younger brothers of homosexual male transsexuals in this study are not consistent with the progressive immunization hypothesis. PMID- 11037087 TI - Social phobia in a population-based female adolescent twin sample: co-morbidity and associated suicide-related symptoms. AB - BACKGROUND: This report attempted to replicate and extend prior work examining social phobia (SP), co-morbid psychiatric illnesses, and the risk of suicidal ideation and suicide attempts incurred by their adolescent sufferers. METHODS: SP, alcohol dependence (ALD) and major depressive disorder (MDD) diagnoses, and suicide-related symptoms, were assessed in a population-based adolescent female twin sample. The differentiation of risks as a function of co-morbidity was explored. A trivariate model was fitted to estimate sharing of genetic and environmental vulnerability between SP and co-morbid disorders. RESULTS: The lifetime prevalence of SP was 16.3 %. Significant risk for co-morbid MDD (OR = 3.2) and ALD (OR = 2.1) was observed. Strong evidence for shared genetic vulnerability between SP and MDD (respective heritabilities 28%, 45%; genetic r = 1.0) was observed with moderate support noted for similar sharing between SP and ALD (genetic r = 0.52, heritability for ALD 63%). SP with co-morbid MDD was associated with elevated risk for ALD and for suicide-related symptoms. CONCLUSIONS: SP is a common illness often followed by co-morbid MDD and ALD. SP with comorbid MDD predicts a substantially elevated risk of ALD and suicide related symptoms, stressing the need for early SP detection. PMID- 11037088 TI - An educational intervention for front-line health professionals in the assessment and management of suicidal patients (The STORM Project). AB - BACKGROUND: Suicide prevention is a health priority in many countries. Improved management of suicide risk may improve suicide prevention. This study aimed to assess the feasibility of health district-wide training in the assessment and management of people at risk of suicide; and to assess the impact of training on assessment and management skills. METHODS: Staff in three health care settings, namely primary care, accident and emergency departments and mental health services (N = 359), were offered suicide risk management training in a district wide programme, using a flexible 'facilitator' approach. The main outcomes were the rate of attendance at training, and changes in suicide risk assessment and management skills following training. RESULTS: It was possible to deliver training to 167 health professionals (47 % of those eligible) during a 6 month training period. This included 95 primary care staff (39%), 21 accident and emergency staff(42%) and 51 mental health staff (78%). Of these, 103 (69%) attended all training. A volunteer sample of 28 staff who underwent training showed improvements in skills in the assessment and management of suicide risk. Satisfaction with training was high. The expected costs of district-wide training, if it were able to produce a 2.5% reduction in the suicide rate, would be 99,747 pound sterling per suicide prevented and 3,391 pound sterling per life year gained. CONCLUSIONS: Training in the assessment and management of suicide risk can be delivered to approximately half the targeted staff in primary care, accident and emergency departments and mental health services. The current training package can improve skills and is well accepted. If it were to produce a modest fall in the suicide rate, such training would be cost-effective. However, a future training programme should develop a broader training package to reach those who will not attend. PMID- 11037089 TI - Suicide and attempted suicide among older adults in Western Australia. AB - BACKGROUND: Suicide rates are high in later life. Risk factors include male sex and depressive illness. This study investigated the relationship between suicidal behaviour and contact with mental health services among the elderly in Western Australia. METHODS: Record linkage was used to obtain records of hospital admissions and mental health service contacts for all suicide attempts and deaths in the period 1980-95. Standardized incidence ratios were calculated for the elderly, general population and people with mental health service contacts. Cox regression was used to evaluated potential risk factors for elderly people who were in contact with mental health services. RESULTS: People over 60 years of age accounted for 15% of suicides and 4.6% of attempted suicides. Suicide rates were 3.3 times higher in males and 4.4 times higher in females when compared to the general population of elderly people. For attempted suicide, the rate was 5.8 times higher in males and 6.6 times higher in females with prior contact with mental health services. Highest risk of suicide was found in patients with diagnoses of affective psychoses (RR = 3.7), adjustment reaction (RR = 3.2) or depressive disorder (RR = 2.8). The diagnosis of cancer was associated with decreased risk of suicide (RR = 3.6) and attempted suicide (RR = 1.9). CONCLUSIONS: Suicide rates are high among the elderly in Western Australia. Suicide is significantly associated with the diagnosis of mood disorder. Suicide attempts are less common, and are associated most strongly with mood and personality disorders. The decreased risk of self-harm behaviour among patients with cancer warrants further investigation. PMID- 11037090 TI - The stability of the factor structure of the General Health Questionnaire. AB - BACKGROUND: Different versions of the General Health Questionnaire (GHQ), including the GHQ-12 and GHQ-28 have been subjected to factor analysis in a variety of countries. The World Health Organization study of psychological disorders in general health care offered the opportunity to investigate the factor structure of both GHQ versions in 15 different centres. METHODS: The factor structures of the GHQ-12 and GHQ-28 extracted by principal component analysis were compared in participating centres. The GHQ-12 was completed by 26,120 patients and 5,273 patients completed the GHQ-28. The factor structure of the GHQ-28 found in Manchester in this study was compared with that found in the earlier study in 1979. RESULTS: For the GHQ-12, substantial factor variation between centres was found. After rotation, two factors expressing depression and social dysfunction could be identified. For the GHQ-28, factor variance was less. In general, the original C (social dysfunction) and D (depression) scales of the GHQ-28 were more stable than the A (somatic symptoms) and B (anxiety) scales. Multiple cross-loadings occurred in both versions of the GHQ suggesting correlation of the extracted factors. In Manchester, the factor structure of the GHQ had changed since its development. Validity as a case detector was not affected by factor variance. CONCLUSIONS: These findings confirm that despite factor variation for the GHQ-12, two domains, depression and social dysfunction, appear across the 15 centres. In the scaled GHQ-28, two of the scales were remarkably robust between the centres. The cross-correlation between the other two subscales, probably reflects the strength of the relationship between anxiety and somatic symptoms existing in different locations. PMID- 11037091 TI - The Personal Health Questionnaire: a new screening instrument for detection of ICD-10 depressive disorders in primary care. AB - BACKGROUND: The next generation of studies on antidepressant drug prescriptions in general practice needs to assess both the patterns of prescription and its appropriateness. This study aimed to assess the performance of the Personal Health Questionnaire (PHQ), a new questionnaire for detecting individuals with ICD- 10 depressive disorders, to be used in association with companion instruments for assessing the 'quality' of antidepressant prescriptions in primary care settings. METHODS: The PHQ was completed by 1,413 primary care attenders (100 were re-tested after 7-14 days) and 139 were selected and interviewed using the SCAN-2 and the 17-item HDRS. All data were analysed using appropriately weighted procedures to control for two-phase sampling design and non-response bias. Individual weights were estimated by logistic regression analysis and trimming strategy. RESULTS: PHQ internal consistency and test-retest on both Likert score and number of symptoms were high. The PHQ discriminated well between individuals with and without depressive disorders. A Likert score > or = 9 provided a good trade-off between sensitivity (0.78) and specificity (0.83). The screening accuracy of the PHQ in detecting subjects likely to benefit from antidepressant drug treatment (SCAN cases with a HDRS total score of 13 or higher) was satisfactory (ROC area 0.87: sensitivity 0.84; specificity 0.78). CONCLUSIONS: The PHQ can be strongly suggested as an accurate and economic screener to identify primary care attenders at high risk of being clinically depressed. However, in order to identify patients requiring antidepressant drug treatment, a second-phase assessment of PHQ high scorers (total score of > or = 10), using the HDRS, is needed. PMID- 11037092 TI - The Reassurance Questionnaire (RQ): psychometric properties of a self-report questionnaire to assess reassurability. AB - BACKGROUND: The aim of this study was to develop a questionnaire that assessed the extent to which patients usually feel reassured by their attending physician. METHODS: The study population consisted of 204 subjects from the general population, 113 general practice patients, 130 general medical out-patients and 183 general medical patients with unexplained physical symptoms participating in an intervention study on the effect of cognitive behavioural therapy. RESULTS: Factor analysis yielded a one-factor solution. The internal consistency was moderate to high and the test-retest reliability was high. The convergent validity of the Reassurance Questionnaire (RQ) was satisfactory to good, but the scores on the RQ did not appear to differentiate between the general population, general practice patients and general medical out-patients. In medical out patients with unexplained physical symptoms, the RQ discriminated well between hypochondriacal and non-hypochondriacal patients. Scores on the RQ tended to be associated with a bad outcome in terms of recovery of presenting symptoms at 1 year follow-up. There was no association between scores on the RQ and frequency of physician contact. In patients with unexplained physical symptoms treated with cognitive behavioural therapy, scores on the RQ decreased over a period of 6 months and 1 year. CONCLUSIONS: The RQ was demonstrated to have psychometrically sound properties and appeared to be a useful instrument to assess reassurability in medical patients. PMID- 11037093 TI - Fatigue rating scales: an empirical comparison. AB - BACKGROUND: There has been limited research comparing the efficacy of different fatigue rating scales for use with individuals with chronic fatigue syndrome (CFS). This investigation explored relationships between two commonly-used fatigue rating scales in CFS research, the Fatigue Scale and the Fatigue Severity Scale. Theoretically, these scales have been described as measuring different aspects of the fatigue construct. The Fatigue Scale was developed as a measure of the severity of specific fatigue-related symptoms, while the Fatigue Severity Scale was designed to assess functional outcomes related to fatigue. METHODS: Associations of these scales with the eight definitional symptoms of CFS and with eight domains of functional disability were examined separately in: (1) an overall sample of individuals with a wide range of fatigue severity and symptomatology; (2) a subsample of individuals with CFS-like symptomatology, and, (3) a subsample of healthy controls. RESULTS: Findings revealed that both scales are appropriate and useful measures of fatigue-related symptomatology and disability within a general population of individuals with varying levels of fatigue. However, the Fatigue Severity Scale appears to represent a more accurate and comprehensive measure of fatigue-related severity, symptomatology, and functional disability for individuals with CFS-like symptomatology. PMID- 11037094 TI - Quality of life impairments associated with diagnostic criteria for traumatic grief. AB - BACKGROUND: This study examined the association between a diagnosis of traumatic grief and quality of life outcomes. METHOD. Sixty-seven widowed persons were interviewed at a median of 4 months after their loss. The multiple regression procedure was used to estimate the effects of a traumatic grief diagnosis on eight quality of life domains, controlling for age, sex, time from loss and diagnoses of major depressive episode and post-traumatic stress disorder. RESULTS: A positive traumatic grief diagnosis was significantly associated with lower social functioning scores, worse mental health scores, and lower energy levels than a negative traumatic grief diagnosis. In each of these domains, traumatic grief was found to be a better predictor of lower scores than either major depressive episode or post-traumatic stress disorder. CONCLUSIONS: The results suggest that a traumatic grief diagnosis is significantly associated with quality of life impairments. These findings provide evidence supporting the criterion validity of the proposed consensus criteria and the newly developed diagnostic interview for traumatic grief the Traumatic Grief Evaluation of Response to Loss (TRGR2L). PMID- 11037095 TI - Eye movement desensitization and reprocessing in the treatment of post-traumatic stress disorder: a review of an emerging therapy. AB - BACKGROUND: Eye Movement Desensitization and Reprocessing (EMDR) is a relatively new form of psychotherapy for post-traumatic stress disorder. We critically reviewed randomized controlled trials of EMDR. METHODS: A wide range of electronic databases and reference lists of articles obtained were searched and relevant experts were consulted. Studies were critically appraised according to established criteria. RESULTS: We found 16 published randomized controlled trials (RCTs) comparing EMDR with alternative psychotherapy treatments, variants of EMDR and with delayed treatment groups. Studies were generally small (mean number of patients = 35) and of variable methodological quality, with only five reporting blinding of outcome assessors to treatment allocation, and in some cases with high loss to follow-up. In most cases EMDR was shown to be effective at reducing symptoms up to 3 months after treatment. In one case benefit was maintained up to 9 months and in another (uncontrolled) follow-up treatment effect was present at 15 months. Two studies suggest that EMDR is as effective as exposure therapies, three claim greater effectiveness in comparison to relaxation training, and three claim superiority over delayed treatment groups. Of the studies examining specific treatment components, two found that treatment with eyes moving was more effective than eyes fixed, while three studies found the two procedures to be of equal effectiveness. CONCLUSION: The evidence in support of EMDR is of limited quality but results are encouraging for this inexpensive, simple therapy. Further research is warranted in larger samples with longer periods of follow-up. PMID- 11037096 TI - Imaging attentional and attributional bias: an fMRI approach to the paranoid delusion. AB - BACKGROUND: The pathophysiology of auditory hallucinations and delusions of control has been elucidated using functional imaging. Despite their clinical importance, there have been few similar attempts to investigate paranoid delusions. We have examined two components of social cognition (attentional and attributional biases) that contribute to the formation and maintenance of paranoid delusions, using functional magnetic resonance imaging (fMRI). METHOD: Normal subjects performed tasks requiring attentional and attributional judgements. We investigated the neural response particularly associated with attention to threatening material relevant to self and with the 'self-serving' attributional bias. RESULTS: The determination of relevance to self of verbal statements of differing emotional valence involved left ventrolateral prefrontal cortex (left inferior frontal gyrus, BA 47), right caudate and right cingulate gyrus (BA 24). Attention to threatening material relevant to self differentially activated a more dorsal region of the left inferior frontal gyrus (BA 44). Internal attributions of events, where the self was viewed as an active intentional agent, involved left precentral gyrus (BA 6) and left middle temporal gyrus (BA 39). Attribution of events in a non 'self-serving' manner required activation of the left precentral gyrus (BA 6). CONCLUSIONS: Anomalous activity or connectivity within these defined regions may account for the attentional or attributional biases subserving paranoid delusion formation. This provides a simple model for paranoid delusion formation that can be tested in patients. PMID- 11037097 TI - The use of cognitive context in schizophrenia: an investigation. AB - BACKGROUND: Cognitive deficits in schizophrenia have recently been ascribed to impaired representation and use of cognitive context. Context is defined as relevant information held temporarily in mind to mediate appropriate but often non-habitual responses. METHODS: Parallel studies in a variety of cognitive domains were designed in order to explore the generality of any schizophrenic deficit in context use. In all of the tasks (a Stroop task, a Continuous Performance Task and a cued spatial location task), we examined how performance was affected by the time for which contextual information must be held in mind, and by whether context or task demands were consistent or varying between trials. It was predicted that manipulation of these variables would produce tests especially sensitive to schizophrenic attentional problems. RESULTS: Predictions were partially confirmed. Although increasing contextual demands failed in most cases to produce disproportionate slowing of performance in patients, error data were largely in line with predictions. At the same time, the data did not suggest a simple unitary context deficit. Instead, different aspects of context--the time over which contextual information must be held in mind and the consistency of context--were differentially important in different tasks. CONCLUSIONS: The cognitive impairments of schizophrenic patients cannot be simply characterized as a generalized context deficit. A more differentiated, if not task specific, picture of schizophrenic deficits is suggested. PMID- 11037098 TI - Attributions of causality, responsibility and blame for positive and negative symptom behaviours in caregivers of persons with schizophrenia. AB - BACKGROUND: Causal, responsibility and blame attributions for positive and negative symptom behaviours were examined in 70 caregivers of persons with schizophrenia. METHODS: The majority of subjects belonged to self-help group organizations. The three types of attributions for positive and negative symptom behaviours were assessed by self-report questionnaires. RESULTS: The extent of patient responsibility did not differ between the two types behaviours. Intentionality and knowledge were equally important in predicting responsibility for positive symptom behaviours, while intent was the most important predictor of responsibility for negative symptom behaviours with the patient capacity playing a significant but minor role. The entailment model was not supported for the two types of behaviours. CONCLUSIONS: Increased attention should be given to responsibility dimensions in assigning moral accountability to the patient. The entailment model should be further explored in problematical caregiving situations. PMID- 11037099 TI - Decentring and distraction reduce overgeneral autobiographical memory in depression. AB - BACKGROUND: Increased recall of categorical autobiographical memories is a phenomenon unique to depression and post-traumatic stress disorder, and is associated with a poor prognosis for depression. Although the elevated recall of categorical memories does not change on remission from depression, recent findings suggest that overgeneral memory may be reduced by cognitive interventions and maintained by rumination. This study tested whether cognitive manipulations could influence the recall of categorical memories in dysphoric participants. METHODS: Forty-eight dysphoric and depressed participants were randomly allocated to rumination or distraction conditions. Before and after the manipulation, participants completed the Autobiographical Memory Test, a standard measure of overgeneral memory. Participants were then randomized to either a 'decentring' question (Socratic questions designed to facilitate viewing moods within a wider perspective) or a control question condition, before completing the Autobiographical Memory Test again. RESULTS: Distraction produced significantly greater decreases in the proportion of memories retrieved that were categorical than rumination. Decentring questions produced significantly greater decreases in the proportion of memories retrieved that were categorical than control questions, with this effect independent of the prior manipulation. CONCLUSIONS: Elevated categorical memory in depression is more modifiable than has been previously assumed; it may reflect the dynamic maintenance of a cognitive style that can be interrupted by brief cognitive interventions. PMID- 11037100 TI - Psychiatric illness predicts poor outcome after surgery for hip fracture: a prospective cohort study. AB - BACKGROUND: Hip fracture is common in the elderly. Previous studies suggest that psychiatric illness is common and predicts poor outcome, but have methodological weaknesses. Further studies are required to address this important issue. METHODS: We prospectively recruited 731 elderly participants with hip fracture in two Leeds hospitals. Psychiatric diagnosis was made within 5 days of surgery using the Geriatric Mental State schedule and other standardized instruments, and data on confounding factors was collected. Main study outcomes were length of hospital stay, and mortality over 6 months after fracture. RESULTS: Fifty-five per cent of participants had cognitive impairment (dementia in 40% and delirium in 15%), 13% had a depressive disorder, 2% had alcohol misuse and 2% had other psychiatric diagnoses. Participants were likely to remain in hospital longer if they suffered from dementia, delirium or depression. The relative risks of mortality over 6 months after hip fracture were increased in dementia and delirium, but not in depression. CONCLUSIONS: Psychiatric illness is common after hip fracture, and has significant effects on important outcomes. This suggests a need for randomized, controlled trials of psychiatric interventions in the elderly hip fracture population. PMID- 11037101 TI - Attachment style in patients with unexplained physical complaints. AB - BACKGROUND: Patients who present with physical symptoms that lack an organic explanation are common, difficult to help and poorly understood. Their medical help-seeking is a form of care-eliciting behaviour and, as such, may be understandable in terms of attachment style. Adult attachment style influences functioning in relationships, and may affect help-seeking behaviour from professional carers such as the family doctor. METHOD: A consecutive sample of 2,042 primary-care attenders completed questionnaires on: the reason for consultation, attribution of symptoms, psychiatric distress (GHQ), somatic distress (BSI), and self-reported adult attachment style (ASQ). Their doctors rated presentations into explained physical, unexplained physical, or psychological. RESULTS: There is a powerful relationship between type of presentation and adult attachment style. Both abnormal attachment and level of psychiatric distress increased significantly from the explained physical group, through the unexplained physical group to the group who presented psychologically. Logistic regression models determined three explanatory variables that made significant independent contributions to presentation type: psychiatric distress, attachment style and symptom attribution. CONCLUSION: Presentation to the doctor with unexplained physical symptoms is associated with both higher levels of psychiatric symptoms and abnormal attachment style when compared to presentations with organic physical symptoms. Patients who present overt psychological symptoms suffer more psychiatric distress and have more abnormal attachment than those presenting physical symptoms (either organically explained or unexplained). Models to explain these findings are discussed. PMID- 11037102 TI - Psychological morbidity among informal caregivers of older people: a 2-year follow-up study. AB - BACKGROUND: It has long been assumed that mental frailty in older care-recipients results in a deterioration in family caregivers' psychological well-being. This hypothesis has not been tested in longitudinal studies. Research about the impact on families of supporting older people in institutions is limited. The present study examined psychological morbidity in informal caregivers of frail older people at home and in institutions. Caregivers were followed up over 2 years. Predictors of psychological morbidity in caregivers and factors related to deterioration in their well-being over time were identified. METHODS: Two-year panel survey of informal caregivers of older care-recipients, using semi structured interview schedules and the 30-item GHQ. Care-recipients were those defined as physically or mentally frail after screening a stratified random sample of people aged > or = 65. Respondents were 276 caregivers of older people living at home at baseline and 47 visitors of those in long-term care at baseline. RESULTS: There were no differences in GHQ scores between caregivers at home and visitors, and no changes in GHQ score over time. The strongest predictors of psychological morbidity at follow-up, and of change in GHQ scores over time, were baseline GHQ score and indicators of subjective well-being. Characteristics of care-recipients, including frailty type (mental or physical), were not significantly associated with changes in caregivers' psychological morbidity. CONCLUSIONS: Mental frailty in care-recipients was not associated with deterioration over time in caregivers' psychological well-being. Caregiver characteristics were stronger than those of care-recipients in predicting psychological morbidity at follow-up and in predicting deteriorating well-being over time. PMID- 11037103 TI - Screening for cognitive impairment in older African-Caribbeans. AB - BACKGROUND: There are increasing numbers of older African-Caribbeans in the United Kingdom. Screening instruments are commonly used in the detection of cognitive impairment, but have not been assessed within this population. This study aimed to develop culturally modified versions of screening instruments for cognitive impairment (Mini-Mental State Examination (MMSE) and Abbreviated Mental Test (AMT)) and to determine their sensitivity and specificity in the diagnosis of dementia. METHODS: The instruments were modified using a process involving a community group of African-Caribbeans and an academic group of health professionals. They were used in a two-stage study involving community resident African-Caribbeans aged 60 years or over in inner-city Manchester, comparing the screening instruments against a computerized diagnostic interview. RESULTS: One hundred and thirty people completed the study. The results for the largest subgroup, the Jamaicans (N = 96) were analysed. Effects of gender, age and education on the MMSE and AMT scores were evaluated. The correlations between the screening instruments and diagnostic interview were highly significant (P < 0.001). At appropriate cut-offs both screening instruments demonstrated high sensitivity and acceptable specificity levels. CONCLUSIONS: A defined process with lay input has assisted in producing culturally modified versions of the MMSE and AMT that perform well compared with a diagnostic interview, if an appropriate cut-off is used. They are easy to administer and acceptable to older African Caribbean people. The results need to be viewed within the limitations of the current study. PMID- 11037104 TI - Absence of interactions between social support and stressful life events in the prediction of major depression and depressive symptomatology in women. AB - BACKGROUND: While high levels of social support (SS) are associated with a decreased risk for major depression (MD) or less depressive symptomatology, and stressful life events (SLEs) have a substantial causal relationship with MD, uncertainty remains as to whether a main-effect or a buffering model best explains the nature of the relationship among SS, MD and SLEs. METHOD: Using two waves of interview data on 2,163 female twin pairs from a population-based twin registry, and discrete time survival analysis with both logistic and linear regression models, we examine the ability of interactions between eight dimensions of SS and 16 categories of stressful life events to predict MD onset and levels of depressive symptomatology. RESULTS: In the presence of a significant effect of a SLE on MD (beta > or = 100), we found evidence for seven interactions out of a possible 93, of which none involved buffering effects. Similarly, examination of depressive symptomatology detected a total of two interactions (both buffering) out of possible 28. We found no evidence, beyond what would be expected by chance, for the existence of buffering effects where either MD or depressive symptomatology was used as the dependent variable. CONCLUSIONS: There is little evidence to suggest the presence of the buffering effect of social support in the face of adverse life events for women. We suggest that it is important to use alternative models (multiplicative and additive) to examine data, to investigate the match between stressors and social resources, and to investigate fully whether detected interactions actually represent a buffering effect. PMID- 11037105 TI - The effects of trauma among kidnap victims in Sardinia, Italy. AB - BACKGROUND: No study to date has investigated the effects of the trauma of being kidnapped for ransom. In the present study, we aimed to assess the general health status and the presence of post-traumatic stress disorder (PTSD) and major depression (MDD) in a sample of kidnap victims. We also focused attention on dissociative experiences and on the development of the Stockholm syndrome during captivity. METHODS: We investigated the traumatic experiences and reported general health status of 24 kidnap victims using a semistructured interview. The Structured Clinical Interview for DSM-IV was used to assess the presence of PTSD and MDD. The Dissociative Experiences Scale was also administered. RESULTS: The lifetime frequency of PTSD and MDD were 45.9% and 37.5% respectively. The Stockholm syndrome had been present in 50% of the sample during captivity. The presence of PTSD can be predicted by the number of violent experiences, whereas the number of humiliating or deprivation experiences predicts the development of the Stockholm syndrome. Subjects with both PTSD and the Stockholm syndrome reported a greater number of physical complaints at the interview. CONCLUSIONS: There is no significant connection between PTSD and the Stockholm syndrome. Both are indices of the severity of the trauma of being kidnapped, but they are associated with different aspects of the traumatic experience. The presence of both syndromes appears to have a detrimental effect on physical health. PMID- 11037106 TI - Informant ratings of cognitive decline in old age: validation against change on cognitive tests over 7 to 8 years. AB - BACKGROUND: Informant questionnaires on cognitive decline are increasingly used as screening tests for dementia. Informants can provide a longitudinal perspective that is not possible with cognitive screening tests administered at one point in time. However, there are limited data on the validity of such questionnaires when judged against longitudinal change on cognitive tests. METHODS: A community sample of elderly people aged > or = 70 was assessed on cognitive tests at baseline and after a follow-up of 7-8 years. The participants were given the Mini-Mental State Examination and tests of episodic memory and mental speed. At follow-up, the short-form of the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE) was completed by informants for 287 of the participants. RESULTS: Elderly people who were rated by informants as having moderate or severe decline had declined significantly on cognitive tests. The IQCODE correlated as highly with cognitive test change scores as these change scores correlated with each other. CONCLUSIONS: The IQCODE is a valid way of assessing cognitive decline when assessment can be carried out only at one point in time. PMID- 11037107 TI - High rates of attempted suicide in Asian women, especially at young ages; investigation of precipitating factors. PMID- 11037108 TI - Characterization of transgenic mouse strains using six human hepatic cytochrome P450 probe substrates. AB - 1. Transgenic mice were evaluated with six human cytochrome P450 (CYP) selective probe substrates, as little is known about their metabolism in the mouse. Mouse strains characterized include C57BL/SJL, FVB/N, mdr 1a/1b (-/-), ob/ob and ACCA. 2. Human CYP probe substrates used for characterization of mouse CYP activities included bufuralol, testosterone, dextromethorphan, phenacetin, diclofenac and S mephenytoin. Activities were compared with those obtained in human liver microsomes and in human recombinant enzyme preparations. All transgenic mouse strains showed similar apparent K(m) with bufuralol, testosterone and dextromethorphan which compared favourably with those observed in human liver microsomes. 3. K(m) for phenacetin O-deethylase and S-mephenytoin 4' hydroxylation were more variable across strains and in some cases demonstrated biphasic kinetics. Phenacetin O-deethylase activity was low in all mouse strains except FVB/N and mdr 1a/1b (-/-). Diclofenac 4-hydroxylation did not occur to any significant extent in the five strains of mouse evaluated here. 4. The findings suggest the validity of using five of the probes for transgenic mouse hepatic CYP characterization and gross comparison with data generated with human CYP. PMID- 11037109 TI - Purification and characterization of oxidoreductases-catalyzing carbonyl reduction of the tobacco-specific nitrosamine 4-methylnitrosamino-1-(3-pyridyl)-1 butanone (NNK) in human liver cytosol. AB - 1. Four enzymes were purified to homogeneity from human liver cytosol and were demonstrated to be responsible for carbonyl reduction of the tobacco-specific nitrosamine 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK). 2. Carbonyl reductase (EC 1.1.1.184), a member of the short-chain dehydrogenase/reductase (SDR) superfamily, was compared with three isoenzymes of the aldo-keto reductase (AKR) superfamily in terms of enzyme kinetics, co-substrate dependence and inhibition pattern. 3. AKR1C1, 1C2 and 1C4, previously designated as dihydrodiol dehydrogenases (DD1, DD2 and DD4), showed lower K(m) (0.2, 0.3 and 0.8 mM respectively) than did carbonyl reductase (7 mM), whereas carbonyl reductase exhibited the highest enzyme efficiency (Vmax/K(m)) for NNK. Multiplication of enzyme efficiencies with the relative quantities of individual enzymes in cytosol resulted in a rough estimate of their contributions to total alcohol metabolite formation. These were approximately 60% for carbonyl reductase, 20% each for AKR1C1 and 1C2, and 1% for AKR1C4. 4. Except for AKR1C4, the enzymes had a strong preference for NADPH over NADH, and the highest activities were measured with an NADPH-regenerating system. Carbonyl reductase activity was extensively inhibited by menadione, rutin and quercitrin, whereas medroxyprogesterone acetate, phenolphthalein and flufenamic acid were potent inhibitors of AKR1C1, 1C2 and 1C4. 5. In conclusion, cytosolic members of the SDR and AKR superfamilies contribute to reductive NNK detoxification in human liver, the enzymes responsible being carbonyl reductase and aldoketo reductases of the AKRIC subfamily. PMID- 11037110 TI - Modulation of rat epididymal gamma-glutamyl transpeptidase by nonylphenols. AB - 1. gamma-Glutamyl-transpeptidase (gamma-GTP), present at low levels in the testis, seminal vesicle, prostate gland and epididymis in rat at 4 days of age, showed rapid developmental increases at the time of weaning. 2. Administration of nonylphenols (NP) to the neonatal male rat pup (from days 1 to 15) impaired the subsequent development of gamma-GTP in the epididymis but not in the testis, seminal vesicles or prostate gland. 3. Single injection of NP to weaned pups at approximately 22 days of age decreased gamma-GTP in the epididymis but not in other male accessory sex organs. This effect was transient, dose-dependent and blocked by the oestrogen receptor-specific antagonist ICI 182,780. 4. Single injection of oestradiol to weaned rat at approximately 22 days of age also decreased gamma-GTP in the epididymis but not in the testis, prostate gland or seminal vesicles. 5. In in vitro assays, NP did not inhibit epididymal gamma-GTP activity even at 100 microM final concentration. Under similar conditions, acivicin, a specific inhibitor for gamma-GTP, showed a dose-dependent inhibition of gamma-GTP activity. 6. It is suggested that NP impair gamma-GTP expression in the epididymis of developing male rat and act in part via the oestrogen receptor. PMID- 11037112 TI - Pharmacokinetics of the M1-agonist talsaclidine in mouse, rat, rabbit and monkey, and extrapolation to man. AB - 1. Talsaclidine is an M1-agonist under development for the treatment of Alzheimer's disease. The aim of the study was to investigate the absorption, distribution, metabolism and excretion (ADME) of single intravenous and oral doses of [14C]-talsaclidine in mouse, rat, rabbit and monkey. Previous data in humans showed that the drug was mainly excreted into the urine as the unchanged parent drug. The hypothesis was tested if animal data of drugs, which are mainly excreted renally, could be extrapolated to human. 2. The apparent volume of distribution at steady-state (V(ss)) was comparable in all animal species (2-5 l x kg(-1)) indicating an extensive distribution of the drug into tissues. The plasma protein binding was low and comparable in all species including man (< or = 7%). Elimination in terms of clearance was rapid-to-moderate depending on the species. The total plasma clearance (Cl) decreased in the order: mouse (128 ml x min(-1) x kg(-1))> rat (73.9) > monkey (10.6). Urinary excretion is the dominant route of excretion (> or = 86%). 3. A good correlation was achieved with human and animal data in allometric scaling of CI and V(ss). This confirms the hypothesis that renal filtration is scalable over the species and, given a comparable protein binding, animal data is predictive for man. PMID- 11037111 TI - CYP isoform induction screening in 96-well plates: use of 7-benzyloxy-4 trifluoromethylcoumarin as a substrate for studies with rat hepatocytes. AB - 1. In this study, 7-benzyloxy-4-trifluoromethylcoumarin (BFC) was evaluated as a substrate to assess the induction of cytochrome P450 (CYP) isoform enzyme activities in rat hepatocytes using a 96-well plate format. 2. BFC was metabolized by both untreated and sodium phenobarbitone (NaPB)-treated rat hepatocytes in a time- and concentration-dependent manner to the highly fluorescent product 7-hydroxy-4-trifluoromethylcoumarin (HFC). 3. HFC was extensively conjugated with D-glucuronic acid and/or sulphate in both untreated and NaPB-treated rat hepatocytes, thus necessitating the inclusion of an enzymatic deconjugation step in the assay procedure. 4. The time-course of induction of 7-ethoxyresorufin metabolism by the CYP1A inducer beta naphthoflavone (BNF), 7-benzyloxyresorufin metabolism by the CYP2B inducer NaPB and BFC metabolism b both BNF and NaPB was studied in rat hepatocytes treated for 24-96 h. The optimal time for induction of metabolism of all three substrates was 72 h, with no medium changes being necessary during this period. 5. The effect of treatment with 0.5-20 microM BNF, 50-2000 microM NaPB, 2-20 microM dexamethasone (DEX), 20-100 microM methylclofenapate (MCP), and 50 and 200 microM isoniazid (ISN) for 72 h on BFC metabolism in cultured rat hepatocytes was studied. BFC metabolism was induced by treatment with BNF, NaPB and MCP, but not with either DEX or ISN. 6. The metabolism of BFC in liver microsomes from the control rat and rat treated with CYP isoform inducers was also studied. BFC metabolism was induced by treatment with NaPB, BNF and DEX. 7. The metabolism of BFC was also studied using microsomes from baculovirus-infected insect cells containing rat cDNA-expressed CYP1A, CYP2B, CYP2C and CYP3A isoforms. Whereas BFC was metabolized to some extent by all the rat cDNA-expressed CYP isoforms examined, at a substrate concentration of 2.5 microM the greatest rates of BFC metabolism were observed with the CYP1A1, CYP1A2 and CYP2B1 preparations. 8. In summary, the results demonstrate that BFC is a good substrate for assessing the induction of CYP1A and CYP2B isoforms in rat hepatocytes in a 96-well plate format. PMID- 11037113 TI - Pharmacokinetics and metabolism of a vitamin D analogue (Seocalcitol) in rat and minipig. AB - 1. Seocalcitol (EB 1089), a vitamin D analogue with strong antiproliferative effects in vitro and in vivo, is presently under clinical evaluation for the systemic treatment of various solid tumours. 2. The aim was to investigate the pharmacokinetics of Seocalcitol after single and multiple oral administration to rat and minipig. Furthermore, the hepatic metabolism of Seocalcitol was studied both in vitro and in vivo. In vitro metabolism was also investigated in human. 3. In rat, the pharmacokinetic profile of Seocalcitol (Cmax, AUC, Tmax, T(1/2)) was the same after single and oral administration. Pharmacokinetics were also demonstrated as dose-independent. The same was more difficult to evaluate in the minipig due to a great variation among individual animals. 4. In the male rat, the serum T(1/2) was 3 h, but in the female rat and minipig (both genders) T(1/2) = 8 h. 5. At Tmax the concentration of Seocalcitol in the liver (both species) was 10-fold higher than the concentration in serum. The major metabolites in the liver were various isomers of 26-hydroxy Seocalcitol, although the concentration of the individual isomers in rat and minipig were not the same. 6. The same metabolites were formed in vitro following incubations with rat, minipig and human S9 fractions. PMID- 11037114 TI - Diltiazem blood pharmacokinetics in the pregnant and non-pregnant rabbit: maternal and foetal tissue levels. AB - 1. The aim was to investigate the pharmacokinetics of diltiazem (DTZ) and its metabolites, deacetyldiltiazem (M1) and N-demethyldiltiazem (MA), in the pregnant rabbit following DTZ intravenous administration. In addition, DTZ tissue distribution in both the non-pregnant and pregnant rabbit and foetuses was also studied. 2. The slope of the alpha- and beta-phases increased slightly in six of the eight pregnant rabbits as compared with the non-pregnant animal, but the other pharmacokinetic parameters that largely determine drug disposition (AUC, V(n), CL) showed no significant differences. 3. MA blood disposition was unaltered by pregnancy. However, all the pharmacokinetic parameters calculated for the deacetylated metabolite of DTZ were significantly modified in the pregnant as compared with the non-pregnant rabbit. 4. DTZ tended to concentrate in most of the tissues examined. Significant differences were observed in the DTZ concentration in the uterus and kidney from the pregnant as compared with the non pregnant rabbit. 5. The findings suggest that DTZ diffuses easily through the placenta, reaching DTZ blood concentrations equivalent to that observed in maternal blood. However, the concentration of DTZ and its metabolites in the selected foetal tissues was either higher (in brain and muscle) or lower than that observed in maternal tissues, suggesting a different tissue affinity and/or a different metabolic activity in the foetuses as compared with the mothers. PMID- 11037115 TI - FELASA recommendations for the education and training of persons carrying out animal experiments (Category B). Report of the Federation of European Laboratory Animal Science Associations Working Group on Education of Persons Carrying out Animal Experiments (Category B) accepted by the FELASA Board of Management. PMID- 11037116 TI - Ethics committee recommendations for laboratory animals in private research in France. AB - Complementary to existing legislation, non-public research companies in France have been working together voluntarily within an organization known as Grice (Interprofessional Working Group on Ethics Committees for Laboratory Animals/Groupe de Reflexion Interprofessionnel sur les Comites d'Ethique appliquee a l'animal de laboratoire) with the objective of creating institutional ethics committees in an effort to promote animal welfare and good scientific procedures. Each company's commitment to the creation of these committees has been expressed by signing the Charter. Each ethics committee is composed of at least three members, including one who is not a scientist; a veterinarian is highly desirable. The committee examines all procedures and protocols involving animals and hands down a favourable or unfavourable opinion, or requests improvements, especially concerning animal well-being. Consensual approval of the protocol is an essential requirement before the purchase or allocation of animals. The committee examines every aspect of laboratory animal housing and care, and inspects all temporary or permanent animal housing facilities. Grice will continue its efforts in relation with public research organizations as well as with groups and in other countries whose objectives are in line with its own. PMID- 11037117 TI - Sufentanil and medetomidine anaesthesia in the rat and its reversal with atipamezole and butorphanol. AB - Injectable anaesthetics are widely used to anaesthetize rats, but recovery times are often prolonged. Reversible anaesthetic regimens have the advantage that animals may be recovered quickly, thus reducing the incidence of postoperative complications such as hypothermia, and also providing a means of treating inadvertent anaesthetic overdose. This study assessed and compared the characteristics of anaesthesia induced with combinations of sufentanil and medetomidine administered as a single subcutaneous or intraperitoneal dose, and reversal with butorphanol and atipamezole. Combinations of sufentanil/medetomidine at 40 microg/150 microg and 50 microg/150 microg/kg administered subcutaneously, and 80 microg/300 microg/kg by intraperitoneal injection were found to produce surgical anaesthesia for 101+/-49, 124+/-45 and 76+/-23 min (means +/- SD) respectively. All three combinations produced marked respiratory depression 30 min after injection (< 50% of resting respiratory rate). Oxygen saturation, measured by pulse oximetry, was < 50% in all groups 30 min following drug administration. Subcutaneous administration is recommended since it resulted in a more reliable and more rapid induction of anaesthesia than intraperitoneal administration. The administration of butorphanol and atipamezole (0.2/0.5 mg/kg s.c.) resulted in a rapid (< 7 min) reversal of anaesthesia and an associated respiratory depression. The induction of anaesthesia with sufentanil/medetomidine and its reversal with a combination of atipamezole and butorphanol is an effective technique for anaesthetizing rats. However, due to the marked respiratory depression and the resulting hypoxia, we recommend that this regimen should only be used in animals which are free from respiratory disease and that oxygen should be provided during anaesthesia. PMID- 11037118 TI - Reduction of isoflurane MAC with buprenorphine and morphine in rats. AB - Preoperative analgesics are being increasingly used to provide analgesia in the intraoperative and postoperative period. Opioids reduce anaesthetic requirements, although the effect varies with the different drug and species. The aim of this work was to determine whether buprenorphine reduces the minimum alveolar concentration (MAC) of isoflurane in a dose-related fashion, and whether this effect is similar to morphine when clinical doses of both drugs are used in the rat. Thirty-six male Wistar rats were anaesthetized with isoflurane, and MAC was determined before and after the administration of either buprenorphine or morphine. MAC of isoflurane was determined from alveolar gas samples when a standard noxious stimulus, in the form of a tail clamp, was applied. The duration and degree of reduction of the MAC of isoflurane were recorded. Basic cardiovascular and respiratory measurements were also recorded. Buprenorphine (10, 30 and 100 microg/kg) and morphine (1, 3 and 10 mg/kg) reduced in a dose dependent fashion the MAC of isoflurane by 15%, 30% and 50%, respectively. Buprenorphine resulted in less cardiovascular and respiratory depression and had a longer-lasting action than morphine. In conclusion, buprenorphine has a dose related isoflurane sparing effect in the rat similar to that caused by morphine at clinical doses of both drugs. PMID- 11037119 TI - The successful use of fentanyl/fluanisone ('Hypnorm') as an anaesthetic for intracranial surgery in neonatal rats. AB - This study reports on the successful use of fentanyl citrate and fluanisone ('Hypnorm') anaesthesia for intracranial surgery in neonatal (7-day-old) rats. Provided the anaesthetic was administered subcutaneously, the animals showed a very high survival rate in the short term (81/85, 95%) and showed no ill effects in the long term. The depth of anaesthesia was sufficient to allow the operation to be carried out without the animal reacting to any painful stimuli. However, the animals did make random movements during the period of surgical anaesthesia which were not related to any painful stimuli. Although these movements did not interfere with the surgery performed here, such movements would interfere with operations requiring greater precision, such as the localized micro-injection of neural tracers. PMID- 11037120 TI - An alternative method to stereotactic inoculation of transplantable brain tumours in large numbers of rats. AB - The rat 9L gliosarcoma brain tumour model has been widely used in brain cancer studies. Intracerebral implantation of the cells in the parietal lobe of the brain has been performed using the stereotactic or freehand inoculation methods. For large numbers of rats, we wished to develop a method more accurate and precise than the freehand method, but less labour intensive than the stereotactic method. A template implantation technique was developed and compared quantitatively with the stereotactic method. Rats were inoculated with either the template or stereotactic method at doses of 1000, 5000, 10000, 20000 or 40000 cells. Results of this comparison showed that the template method is precise and accurate for tumour placement within the brain cortex, and decreases labour requirements. Mean survival rates between groups were not significantly different at doses of 5000, 20000 or 40000 cells inoculated. Significance was seen at the low dose of 1000 cells (P < 0.001). This was attributable to an absence of tumour growth in five of six stereotactic rats in this group. Significance was also seen at the 10000 dose level (P < 0.05) with the stereotactic rats again surviving longer than the template rats. However, in this case all the stereotactic rats had tumour growth. Brain weights did not differ significantly between groups, except at the 1000 dose level where no growth of tumour occurred in five of the six stereotactic animals. Body weight gain within one week following surgery did not differ significantly between any of the groups at alpha = 0.05. Studies on rat cadavers showed no statistical difference in placement measurements between the stereotactic and template methods. These results indicate that the template method for intracerebrally implanting tumour cells in rats provides a precise, accurate and rapid procedure that maximizes reproducibility with a significant reduction in labour requirements, when compared with the conventional stereotactic methodology. PMID- 11037121 TI - Telemetric monitoring of blood pressure in freely moving mice: a preliminary study. AB - This paper describes for the first time the possibility for recording the systolic pressure (SP), diastolic pressure (DP), and the mean arterial pressure (MAP) as well as the heart rate (HR) and locomotor activity (LA) in freely moving mice, using a commercially available telemetry and data acquisition system. The system comprises a new, small radio-telemetry transmitter implanted in the peritoneal cavity, a receiver board placed underneath the home cage, a multiplexer and a computer-based data acquisition system. The signals from the receiver were consolidated by the multiplexer and were stored and analysed by the computer. The telemetered pressure signals (absolute pressure) were corrected automatically for changes in atmospheric pressure measured by an ambient pressure monitor. The effects of implantation on animal behaviour, and, after the animals had recovered, the effects of handling on the SP, DP, MAP and HR were examined. The radio-telemetry system for recording the SP, DP, MAP and HR provides an accurate and reliable method for monitoring the direct effects of handling on SP, DP, MAP and HR. In addition, by using this new blood pressure (BP) transmitter, we maintain that BP measurements in freely moving mice are more efficient, reliable, and less labour-intensive than the measurement techniques described in the literature thus far. PMID- 11037122 TI - Comparison of assays for antibodies to Encephalitozoon cuniculi in rabbits. AB - Two indirect immunofluorescence (IIF) assays, two enzyme-linked immunosorbent assays (ELISAs) and the carbon immunoassay (CIA) for determination of antibodies to Encephalitozoon cuniculi were compared using 210 sera of rabbits, 135 of which originated from seven infected colonies, while 75 originated from four uninfected colonies. There was no evidence of a difference between the different assays with respect to the number of positive sera. There was a clear correlation between the quantitative response measured by IIF and CIA and the other assays, and between both IIF tests, while no such correlation was found in the quantitative response measured by ELISAs, which might be explained by the less quantitative nature of the ELISA. Therefore quantitative determination of antibodies to E. cuniculi should be performed by IIF and not by ELISA. The nosographic sensitivities N1 and specificities N2 of the assays were > or = 0.94 and > or = 0.97 respectively. Small differences in N1 and N2 between the assays, although not statistically significant, were responsible for differences in the calculated predictive values of a positive test and of a negative test. As expected, the magnitude of these differences depended on the fraction of positive sera sampled from a given colony. There was strong evidence of such a difference between the fraction of positive sera found in different colonies, but the sample size from some colonies was too small to allow any conclusion, whether this was due to differences in the prevalences of the infection in the colonies or something else. We conclude that any of the assays will be suitable for the routine health monitoring of laboratory rabbit colonies for E. cuniculi infection, as recommended by the Federation of European Laboratory Animal Science Associations. PMID- 11037123 TI - Differential susceptibility to Brugia pahangi infection in Mongolian gerbils (Meriones unguiculatus) of different coat colour. AB - The influence of intraspecific host variables on the response to parasitic infections is an important aspect of host-parasite relationships, yet little is known about this aspect of filariasis for lack of a model. This study presents coat colour mutants of the Mongolian gerbil (Meriones unguiculatus) as potential new models for research into the effects of host genetic variation on response to filarial infection. Peak level of microfilaraemia, eosinophil response, body weight and degree of splenomegaly in gerbils infected with Brugia pahangi varied with agouti, albino, and black coat colour. These results suggested that coat colour-related genes might influence host immune response to developmental stages of the parasite and eosinophil-mediated reaction might cause host damage. PMID- 11037124 TI - An intra-articular method for assessing topical application to a synovial joint. AB - Intra-articular injection of drugs is increasingly used in human medicines. We report a method for the direct administration of a test substance to the synovial fluid of the canine stifle joint. This method caused little distress or pathology, making it suitable for pre-clinical assessment of new drugs in dogs and other species. PMID- 11037125 TI - Feeding sugar overnight maintains metabolic homeostasis in rats and is preferable to overnight starvation. AB - Rats are often starved overnight for many different reasons. Overnight starvation causes loss of body and liver weights, depletion of liver glycogen, decrease of blood glucose and loss of amino acids because of gluconeogenesis. Providing pure sucrose cubes as the sole overnight nutrient is a simple, inexpensive way to empty the gastrointestinal (GI) tract, while minimizing liver changes and preventing decrease of blood glucose and loss of amino acids. Adding sugars to the overnight drinking water as the sole nutrient has the same beneficial effects, provided the type of sugar and its concentration allow for sufficient intake and provided hyponatremia is avoided. Feeding sucrose cubes or sugar solutions will empty the gastrointestinal tract as effectively as starvation. In all instances, simple precautions against coprophagy and pica should be taken in order to secure optimal benefit. PMID- 11037126 TI - Effects of various levels of dietary Artemisia abyssinica leaves on rats. AB - Artemisia abyssinica leaves, a traditional medicine for the treatment of various disorders, were fed to male Wistar rats at 2% and 10% of the standard diet for 6 weeks. A 2% A. abyssinica leaf diet was not toxic to rats. Depression in growth, hepatopathy and nephropathy were observed in rats fed a diet containing 10% of A. abyssinica leaves. These findings were accompanied by leukopenia, anaemia and alterations of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and gamma glutamyl transferase (GGT) activities with changes in concentrations of total protein, albumin, cholesterol and urea. PMID- 11037127 TI - Preferences of mice, Mus musculus, for different types of running wheel. AB - Mice are increasingly used in research. In particular, their wheel running is often used as a measure of activity, and as a marker of phase of circadian rhythms. Learning about the preferences of mice for different types of wheel may improve their welfare and suggest ways of increasing activity levels. Mice, Mus musculus, were given a choice between different types of running wheel by putting them in cages equipped with two wheels. Strong preferences were shown for wheels with a plastic mesh flooring, rather than the standard metal rods only. The mesh was even preferred over a solid base, although this effect was not seen in mice that had been given access only to wheels with the solid base immediately prior to the choice test. Small diameter wheels, sometimes sold as mouse wheels, were preferred less than standard-sized wheels with rods. The results suggest that types of running wheel often used in laboratories can be improved by considering the animals' preferences. The types of wheel tested here are easy to maintain and entail little additional cost, while increasing the mouse's interest in running and exercise. PMID- 11037129 TI - New insights into symbiotic associations between ants and bacteria. AB - Many ants live in complex mutualistic or parasitic relationships with other insects or plants, some of which are classical examples of the mutual benefit of symbiosis. However, only in the past few years have new insights into the symbiosis of ants and microorganisms been reported. Examples are the symbiosis of ants of the genus Camponotus with intracellular bacteria present in their midgut, and the tripartite relationship of ants of the tribe Attini with an extracellular bacterium essential for the cultivation of their fungus gardens. The analysis of the parasitic and mutualistic interactions of these organisms will allow interesting insights into the evolution of symbiosis and possibly lead to novel strategies of pest control. PMID- 11037128 TI - A tale of two genomes: resolution of dimeric chromosomes in Escherichia coli and Bacillus subtilis. AB - Dimeric chromosomes can be formed during replication of circular bacterial chromosomes by an odd number of homologous recombination events between sister chromosomes. In the absence of a compensating recombination reaction such dimers cannot be segregated from each other as the cell divides. This review highlights the shared and divergent mechanisms employed by Escherichia coli and Bacillus subtilis in their effort to resolve and partition dimeric chromosomes safely. In particular, we discuss the Xer-type recombinases, RecA, FtsK/SpoIIIE, and dif. PMID- 11037130 TI - Oligonucleotide probe for the visualization of Escherichia coli/Escherichia fergusonii cells by in situ hybridization: specificity and potential applications. AB - There are several occasions when enumeration of Escherichia coli cells is needed. These include examination of urine specimens and water or food samples. Present methods rely on growth in more or less selective media (colony-forming units on agar or the most probable number method using liquid media). Unfortunately, no really selective medium with 100% efficiency of plating is available for E. coli. A 24-mer oligonucleotide probe (Colinsitu), complementary to a piece of 16S ribosomal ribonucleic acid, has been tested for specifically visualizing E. coli cells by in situ hybridization and epifluorescence microscopy. The fluorescent dye-labeled probe was able to stain cells of E. coli, Shigella spp. and E. fergusonii. Shigella spp. are known to belong to the E. coli genomospecies and E. fergusonii is the nomenspecies closest to E. coli by DNA-DNA hybridization. The probe did not stain any strain of 169 other genomospecies of the family Enterobacteriaceae or of a few other species frequently encountered in the environment. Revivification without cell division allowed the visualization of E. coli cells in contaminated water. In situ hybridization using the Colinsitu probe is a potential tool for the confirmation of (atypical) E. coli in reference centers and the rapid (3-6 h) detection and enumeration of E. coli in urine specimens, contaminated water and food. More work is needed to include in situ hybridization in laboratory routine. PMID- 11037131 TI - Identification of Escherichia coli flagellar types by restriction of the amplified fliC gene. AB - A total of 182 strains of Escherichia coli (133 reference strains, 22 clinical strains, nine nonmotile strains and 18 strains derived from K-12) were characterized by HhaI restriction of the amplified flagellin gene (fliC). The amplified fliC product was a single band between 0.9 and 2.6 kbp. With the collection of reference strains which represented 48 flagellar types (H-types), a total of 62 patterns (F-types) were observed after HhaI restriction. A single F type was associated with each of 39 H-types and more than one F-type was associated with the other nine H-types. Antigenically related H-types 12 and 45 gave a single F-type. The determination of HhaI-fliC F-types could allow deduction of all H-types and subdivision of some of these. Application of this identification system to 22 E. coli clinical isolates yielded nine F-patterns and the deduced H-types were confirmed by serotyping in all cases. Nine nonmotile strains were studied and their F-types were also identified. The proposed determination of fliC restriction patterns should be helpful for epidemiological studies. PMID- 11037132 TI - Construction of chromosomal integrants of Bacillus sphaericus 2362 by conjugation with Escherichia coli. AB - IncP-based plasmids conjugated between Escherichia coli and mosquitocidal strains of Bacillus sphaericus at frequencies of 10(-7) to 10(-9) per recipient. Plasmid transfer was most efficient when a restriction-deficient strain of B. sphaericus 2362 (serotype 5a5b) was used as recipient and was least efficient with recipients from serotypes 1a and 2a2b. A deleted version of the cryptic locus 'gene 80' from strain 2362 was cloned into the suicide vector pMTL30, which could not replicate in B. sphaericus to provide a site for chromosomal integration. Conjugational transfer from E. coli and integration into the B. sphaericus recipient chromosome was achieved with this construct. The coding region of the cry11A gene from Bacillus thuringiensis subsp. israelensis was PCR-amplified and fused to the promoter of the crystal protein (Bin) gene of B. sphaericus 2362. This construct was cloned into the integrative vector, conjugated with B. sphaericus 2362 and chromosomal integrants were recovered which harboured the cry11A gene. The fusion gene was efficiently transcribed in the recombinant host, but cells failed to accumulate appreciable amounts of Cry11A toxin. This system offers a simple and efficient means of transferring plasmids into B. sphaericus and obtaining chromosomal integration for strain construction and gene analysis. PMID- 11037133 TI - Diphtheria toxin NAD affinity and ADP ribosyltransferase activity are reduced at tryptophan 153 substitutions for alanine or phenylalanine. AB - Previous studies on chemical modifications of diphtheria toxin (DT) fragment A have suggested that the Trp153 amino acid residue is essential for the ADP ribosylation of elongation factor 2. We verified this experimentally after replacing Trp153 by Phe or Ala residues through in vitro mutagenesis of a cloned toxin gene fragment. Each of the mutant fragment A forms were found to reveal a reduced ADP ribosyl transferase (ADPRT) activity as well as lower affinity for NAD. Both ADPRT activity and NAD affinity of DT fragment A were only partially destroyed by nearly synonymous Trp153 ==> Phe153 substitution, but dramatically destroyed by Ala153 substitution. At the same time, each of the mutant fragment A forms appeared to be thermostable, suggesting that the mutations do not dramatically destroy the structure of the protein. These results clearly demonstrate that Trp153 is not highly specific for DT fragment A structure maintenance, but is highly specific for the key toxin functions such as ADP ribosylation of elongation factor 2 and NAD binding. We suggest that the Trp153 role in DT functioning may be that of binding the ribose moiety of NAD, which is crucial for DT catalytic activity and hence for toxicity. PMID- 11037134 TI - Effects of global regulatory proteins and environmental conditions on fimbrial gene expression of F165(1) and F165(2) produced by Escherichia coli causing septicaemia in pigs. AB - Escherichia coli O115:F165 strains are associated with septicaemia in young pigs and possess at least two types of fimbriae. F165(1) fimbriae belong to the P fimbrial family and F165(2) fimbriae belong to the S fimbrial family. Regulatory regions of foo (F165(1)) and fot (F165(2)) fimbrial gene clusters from wild-type strain 4787 were sequenced and characterised. Expression of F165(1) and F165(2) fimbrial genes was analysed by using lacZ and/or luxAB as reporter genes under the control of the native fimbrial promoters. Differential expression of fimbrial genes was observed. Global regulatory mechanisms such as catabolite repression, leucine-responsive regulatory protein (Lrp), methylation and DNA supercoiling were demonstrated to influence foo and fot expression. foo and fot expression was optimal at 37 degrees C and under aerobic conditions. Expression of foo was higher on minimal medium, whereas fot expression was higher on complex Luria Bertani medium. This could reflect an in vivo differential expression. PMID- 11037135 TI - Salmonella flagellin fused with a linear epitope of colonization factor antigen I (CFA/I) can prime antibody responses against homologous and heterologous fimbriae of enterotoxigenic Escherichia coli. AB - In this work, a 15-amino-acid-long peptide derived from the enterotoxigenic Escherichia coli CFA/I fimbria (11VDPVIDLLQADGNAL25) was genetically fused to the Salmonella flagellin and used to prime and boost serum antibody responses (IgG) against homologous (CFA/I) and heterologous (CS1) colonization factors (CFs) in BALB/c mice. Antibodies raised against the hybrid flagellin (Fla II) cross reacted with CFA/I, CS1, CS2, and PCFO166 but not with CS4. Parenteral administration of Fla II primed antibody responses against both CFA/I and CS1 but boosted IgG responses only against CFA/I. These findings confirm that linear epitopes derived from the CFA/I fimbria can prime antibody responses against homologous and heterologous CFs and indicate that Salmonella flagellin represents a potential carrier for the development of broad-range peptide-based anti colonization ETEC vaccines. PMID- 11037136 TI - Genotypic characterization of strains of commercial wine yeasts by tetrad analysis. AB - The objective of this work was to use tetrad analysis to define the genotypes of a number of commercially available wine yeasts for a range of characteristics related to wine making. The levels of sporulation and spore viability of 13 wine yeasts were determined. Sporulation was very low in one strain and varied from low to high in the other 12 strains. Spore viability of these 12 strains varied from 0-95% and this range was comparable to a large sample of naturally-occurring wine strains. Colonies from viable spores, predominantly from 4-spored asci, from 11 strains were characterized for the ten traits: homothallism/heterothallism, fermentation of sucrose, galactose, maltose; growth on glycerol (nonfermentable); slow growth on glucose and glycerol; level of sulfide production; copper resistance; putative presence of a recessive lethal mutation (inviability of at least two spores/tetrad); yellow pigment (in colonies) on sugar media. The number of heterozygosities for these ten characteristics varied from zero to seven in 11 strains, and eight strains were genetically distinct. Another three strains, distinct from these eight strains, were identical for the ten characteristics and also equivalent for the levels of sporulation and spore viability. Although these three strains are marketed under different designations, there is a strong probability that they were derived from a common ancestral strain. The genotypic characterization of these 11 strains constitutes an important foundation for their identification and their use in breeding programs. PMID- 11037137 TI - Levels of neurotransmitter amino acids in the cerebrospinal fluid of patients with acute ischemic insult. AB - The dynamics of excitatory (glutamate, aspartate) and inhibitory (GABA, glycine) neurotransmitter amino acid contents in the cerebrospinal fluid were studied in 110 patients with hemispheric ischemic insult. These studies revealed significant increases in the levels of glutamate and aspartate in the first six hours of illness, and the level and duration of these changes correlated with the severity of the insult. Peak GABA and glycine levels were seen at the end of the first day after strokes, reflecting the delayed activation of the mechanisms of protective inhibition. The insufficiency of GABAergic mediation in strokes located in the hemispheres to a significant extent mirrored the severity of clinical features and the potential of restorative processes. Early significant biochemical criteria were identified for objective assessment of the severity of brain ischemia, and these had prognostic value for the course and outcome of strokes. The most unfavorable prognostic signs were the presence of low (or undetectable) GABA levels in the first days after insult and progressive increases in aspartate levels to the third day on the background of sharp reductions in glutamate levels (after initial elevation on the first day). PMID- 11037139 TI - Electron microscopic characteristics of the dorsomedial nucleus of the amygdaloid body of the brain. PMID- 11037138 TI - Mitochondrial megaconia and pleioconia in the rat brain as possible adaptive reactions in conditions of lethal radiation and radiomodified lesions. PMID- 11037140 TI - Proliferative zones in the epithelium of the choroid plexuses of the human embryo brain. PMID- 11037141 TI - Changes in the topography of a number of outer membrane proteins in cultured neurons in conditions of selective lesioning of different elements of the cytoskeleton with neurotoxins. PMID- 11037142 TI - The morphogenesis of Mauthner neurons in tadpoles of the common frog after early unilateral enucleation of the eye. PMID- 11037143 TI - Localization of NO synthase in Lugaro cells and the mechanisms of NO-ergic interaction between inhibitory interneurons in the rabbit cerebellum. PMID- 11037144 TI - The use of antioxidants to prevent glutamate-induced derangement of calcium ion metabolism in rat cerebral cortex synaptosomes. AB - Glutamate is shown to induce increases in intracellular Ca2+ concentrations ([Ca2+]i), increases in 45Ca2+ influx, decreases in the activity of Na+,K+-ATPase activity, and activation of the Na+/Ca2+ exchanger in rat cerebral cortex synaptosomes. NMDA receptor antagonists virtually prevented these effects. Preincubation of synaptosomes with alpha-tocopherol, superoxide dismutase, and ganglioside GM1 normalized [Ca2+]i, 45Ca2+ influx, and Na+,K+-ATPase activity in rat cerebral cortex synaptosomes exposed to glutamate. Glutamate and GM1 activated the Na+/K+ exchanger, and their effects were additive. Calcium ions entering cerebral cortex nerve cells via NMDA receptors during exposure to high glutamate concentrations appeared to be only the trigger for the processes activating free-radical reactions. Activation of these reactions led to increases in Ca2+ influx into cells, decreases in Na+,K+-ATPase activity, and significant increases in [Ca2+]i, though this could be prevented by antioxidants and gangliosides. PMID- 11037145 TI - The involvement of glutamatergic transmission in the mechanism of movement disorders induced by reversive rotation of white mice. AB - The ability of the selective non-competitive NMDA receptor blocker MK-801 and a series of new glutamate antagonists --the adamantane derivatives IEM-1754 and IEM 1857 and phencyclidine (IEM-1925)--to prevent movement disorders induced by reversive rotation in mice was studied. l.p. MK-801 at a dose of 0.15 ml and IEM 1754 at a dose of 5.0 mg/kg prevented the development of akinesia in response to reversive rotation as effectively as scopolamine, a known agent which provides effective prophylaxis for movement diseases. IEM-1857, the quaternary analog of IEM-1754, was not effective. IEM-1925 significantly increased the responses of mice to reversive rotation, possibly because of its high activity in relation to other subtypes of glutamate receptors. These data provide evidence for the involvement of glutamatergic transmission in the mechanism of movement disorders of vestibular origin. PMID- 11037146 TI - Colocalization of neurotransmitters in presynaptic boutons of inhibitory synapses in the lamprey spinal cord. AB - Electron-microscopic immunocytochemical studies were performed to detect GABA and glycine immunoreactivity in presynaptic axon terminals in the central gray matter of the spinal cord of the lamprey Lampetra fluviatilis. The immunopositive presynaptic terminals contacting identified dendrites of motoneurons and unidentified postsynaptic profiles included terminals immunopositive for GABA only (44%) and glycine only (26%), as well as terminals containing GABA and glycine (30%). Glycine-immunopositive presynaptic terminals contained flattened synaptic vesicles. Large synaptic vesicles with dense cores were present along with classical synaptic vesicles in 74% of GABA-immunopositive boutons. PMID- 11037147 TI - A new class of agonists and antagonists of N-methyl-D-aspartic acid receptors: derivatives of imidazole-4,5- and pyrazole-3,4-dicarboxylic acids. AB - Studies of imidazole-4,5- and pyrazole-3,4-dicarboxylic acid derivatives revealed a number of new agonists and antagonists of N-methyl-D-aspartic acid (NMDA) receptors. Studies were based on whole-cell patch-clamp methods applied to rat hippocampus pyramidal cells. Increases in the lipophilicity of the environment of the nitrogen atom, keeping the distance between the terminal acid functions constant, led to a weakening of NMDA antagonism and increases in NMDA antagonism. Increases in the lipophilicity around the nitrogen atom could also lead to less of selectivity in the interaction with NMDA receptors and the appearance of non NMDA antagonist properties. PMID- 11037148 TI - Characteristics of cholinoreceptors on identified TAN neurons of the ground snail Achatina fulica. AB - The characteristics of cholinoreceptors located on neurons TAN1, TAN2, and TAN3 of the ground snail Achatina fulica were studied by incubation of the central ganglia in a bath with cholinotropic preparations during intracellular recording of background neuron spike activity. Acetylcholine, nicotine, the selective n cholinoreceptor agonist suberyldicholine, and the selective n-cholinoreceptor agonist 5-methylfurmethide concentration-dependently inhibited background spike activity to the level of complete blockade at concentrations of 500 microM. The m cholinoblocker metamizil (500 microM) completely prevented the inhibitory activity of concentrations of 5-methylfurmethide of up to 500 microM. The central n-cholinoblocker etherophen (500 microM) completely blocked the inhibitory activity of 500 microM suberyldicholine. However, metamizil and etherophen added separately only partially decreased the inhibitory effects of acetylcholine but could not completely block the effect of acetylcholine. At the same time, mixtures of metamizil and etherophen (500 microM each) completely blocked the inhibition of background spike activity induced by acetylcholine. These results show that both classes of cholinoreceptors act on TAN neurons in the same direction. PMID- 11037149 TI - Involvement of the striatum in the central regulation of the hormonal functions of the gonads. AB - Studies reported here show that intrastriatal administration of corticoliberin to rats decreases the blood testosterone level. However, in conditions of chemical deficiency of dopaminergic transmission in the dorsal striatum induced by injection of 6-hydroxydopamine, the effect of this neurohormone did not appear. It is concluded that extrahypothalamic corticoliberin is involved in regulating the hormonal reproductive system acting via dopaminergic mechanisms. PMID- 11037150 TI - The effects of sodium nitroprusside on mediator release and the functional properties of postsynaptic membranes in the neuromuscular synapse. AB - Experiments on neuromuscular preparations of frog skin- thoracic muscle and sartorius muscle, using extracellular recording and two-electrode clamping of the muscle fiber membrane potential, were used to study the effects of the nitric oxide donor sodium nitroprusside on endplate currents. At a concentration of 100 microM, sodium nitroprusside sharply decreased the amplitude and quantum composition of the endplate currents, and also decreased the miniature endplate current frequency. The amplitude-time characteristics of miniature endplate currents, the voltage-dependent amplitude, and the decay time constant of miniature endplate currents did not change as compared with controls. However, unlike the situation with other secretion inhibitors, the decrease in endplate current amplitude was not accompanied by increased facilitation in response to rhythmic stimulation or changes in postsynaptic potentiation in conditions of application of pairs of stimuli to muscles. The suppression of acetylcholine secretion was not seen with inactivated sodium nitroprusside solution. These results provide evidence that nitric oxide can be a powerful inhibitor of both spontaneous and evoked transmitter secretion in the neuromuscular synapse, and that this is accompanied by decreases in the efficiency of presynaptic forms of short-term plasticity, while the functional characteristics of the postsynaptic membrane remain unchanged. PMID- 11037151 TI - Intrasynaptic ephaptic feedback in central synapses. AB - Electrophysiological laboratory studies on rat visual cortex and hippocampus slices are reviewed. The aim was to confirm the existence of positive feedback in central synapses operating by an electrical (ephaptic) mechanism, as suggested by Byzov. Byzov's hypothesis holds that artificial hyperpolarization of the postsynaptic membrane potential should increase the amplitude of the excitatory postsynaptic current (EPSC) and potential (EPSP) in some central synapses not only by means of increases in the electromotive force (EMF). but also by means of increases in transmitter release from the presynaptic apparatus. Some experiments showed that hyperpolarization altered the parameters of presynaptic transmitter release, i.e., the quantity of "failed" responses N0, the coefficient of variation CV, and the quantum composition m of minimal EPSC and EPSP. The effect was particularly marked for EPSP in giant synapses formed by mossy fibers on neurons in field CA3. "Supralinear" functions were observed for these synapses in the relationship between EPSC amplitude and membrane potential in conditions of hyperpolarization of membrane potentials and in the relationship between presynaptic paired-stimulus facilitation and membrane potential. All of these "non-classical" effects disappeared when summed rather than minimal EPSC were evoked. The results are in agreement with computer experiments based on the Byzov model and are regarded as support for Byzov's hypothesis. Regardless of their explanation, the data obtained here demonstrate a new feedback mechanism for central synapses, which allows the postsynaptic neuron to control the efficiency of some synapses via changes in membrane potential. This mechanism can significantly increase the efficiency of large ("perforated") synapses and explains the increase in the number of this type of synapse after various experimental manipulations, such as those inducing long-term potentiation or forming conditioned reflexes. PMID- 11037152 TI - Properties of derived cochlear action potentials in forward tonal masking in guinea pigs. AB - Chronic experiments on guinea pigs were used to study the characteristics of cochlear nerve action potentials and the derived potential in conditions of forward masking. The auditory nerve action potential was recorded from the round window of the cochlea. The derived potential was obtained by subtracting the action potential recorded with masking from the response obtained in silence. These studies showed that the derived potential showed higher sensitivity to masking than the traditional measure of masking, i.e., the decrement in the amplitude of the auditory nerve action potential. The derived potential reflects not only changes in the action potential during masking, but also changes in its shape. Differences between the derived potential and the action potential decrement provide evidence that the amplitude and time changes in the derived potential provide more complete information on the nature of activity in the auditory fibers. PMID- 11037154 TI - Characteristics of women with cosmetic breast augmentation surgery compared with breast reduction surgery patients and women in the general population of Sweden. AB - To determine whether women with cosmetic breast implants have distinct demographic, lifestyle, and reproductive characteristics that put them at increased risk for subsequent morbidity, the authors compared 1,369 such women to 2,211 women who had undergone breast reduction surgery, and to a random sample of 49,262 women from the general population of Sweden. Information was collected through self-administered questionnaires, and comparisons were made using the prevalence odds ratio. Women with cosmetic breast implants were significantly (p <0.05) more likely to be current smokers, have a lower body mass index, have had a prematurely terminated pregnancy (induced abortion or miscarriage), and have had fewer live births than either women who underwent breast reduction or women from the general population. Type of implant (silicone gel or saline) did not modify the associations. Regardless of the comparison group used, studies of the health effects of breast implants need to consider that women who undergo cosmetic breast implantation have certain distinct characteristics. PMID- 11037153 TI - Image features selected by neurons of the cat primary visual cortex. AB - The sensitivity of neurons in field 17 of the visual cortex in cats to cross shaped, Y-shaped, and star-shaped figures flashing in the receptive field was studied. About 40% of the neurons studied (114 of 289) were found to generate large responses (with an average response factor of 3.06 +/- 0.32) to one of the figures flashing in the center of the receptive field, as compared with the responses produced to a single bar in the optimal orientation. Most of these neurons (72%) were selectively sensitive to the shape and orientation of figures; the remainder demonstrated some degree of tuning invariance to these properties. The latent periods of responses to figures were usually shorter than those of responses to bars. Tuning parameters for bars and figures were generally related: neurons with acute orientational tuning to a bar were usually highly selective to both the configuration and the orientation people figures. Separate or combined stimulation with crosses in the center and near periphery of the receptive fields demonstrated summation, antagonism, or the lack of any interaction between these zones in producing sensitivity to crosses. Local blockade of intracortical GABAergic inhibition by microiontophoretic application of bicuculline showed that in one third of the neurons studied, sensitivity to figures was generated or enhanced by inhibition in normal conditions, while one third of cells showed suppression by inhibition, and sensitivity in the remainder was independent of inhibition. These data show that reconsideration of existing concepts of the role of field 17 in selecting only first-order shape features of images (i.e., the orientations of single lines) is needed, since almost half the neurons in the cat primary visual cortex can efficiently detect second-order features (angles and line intersections). PMID- 11037155 TI - Using the pulsed dye laser to influence scar formation after breast reduction surgery: a preliminary report. AB - Many patients undergoing bilateral breast reduction surgery develop problems. Foremost among these problems is scar hypertrophy and its attendant symptoms. Although there is as yet no firm prognostic indicator for hypertrophy, scars that become hypertrophic often have a particular blood vessel pattern, observable by transcutaneous microscopy, showing vessels that lay transversely across the incision line with minimal crosslinks between them. Hypertrophic scars that develop in incision lines become wide, with the final width of the scar dependent on the maximum thickness during the growth stages before maturation and resolution. Close monitoring of scars forming in the incision line using the transcutaneous microscope detected this aligned vessel pattern before overt hypertrophy was seen. Use of the Pulsed Dye Laser caused disruption in the vessel pattern, appeared to inhibit additional hypertrophic development, and promoted early maturation of scars. PMID- 11037156 TI - The use of external ultrasound combined with superficial subdermal liposuction. AB - Superficial subdermal liposuction (SSL) was presented by the authors as an evolution in traditional liposuction in 1989. It allows one to treat the superficial fat to obtain better skin retraction during every liposuction procedure. The use of ultrasound (US) in liposuction began 10 years ago as the direct application of this energy to the adipose tissue using a probe to obtain the selective destruction of fat, followed by its aspiration. This technique gave rise to many complications and thus many concerns. The external delivery of US has been proposed to overcome some of the drawbacks associated with this procedure. The authors investigate the role of external ultrasound (EU) used in conjunction with SSL. When comparing EU and SSL with SSL alone, pretreatment with EU results in less bleeding during liposuction, aspirated fat that is clearer and more oily, and less bruising and swelling during the early postoperative period. EU is an ideal complementary procedure to SSL because the crumbling of the fat induced by US permits a more uniform aspiration of the subdermal fat layer, making skin retraction more even and effective. PMID- 11037157 TI - Hazards of piercing and facial body art: a report of three patients and literature review. AB - Piercing has become a widespread fashion trend in Western industrialized nations within recent years. The invasive application of ornaments through cutaneous and mucosal surfaces enables the penetration of various pathogens into subcutaneous tissue. The authors describe the hazards of piercing and facial body art as they apply to 3 patients. Perichondrial auricular abscess, granulomatous perichondritis of the nasal ala, and embedding of a stud in the lower lip were the respective diagnoses. Literature was reviewed for the cultural origins and current practices of piercing, the legal background of piercing as a business, typical medical complications, and treatment recommendations. Numerous communications have been published on medical complications of piercing. The patients presented and the review of the literature illustrate that piercing is not a harmless fashion and that regulations of piercing as a business seem desirable to prevent further complications. PMID- 11037158 TI - Endoscopic-assisted harvest of the temporoparietal fascial flap. AB - Elevation of the temporoparietal fascial flap by conventional T or Y incisions in the temporal region frequently leaves conspicuous scarring, hair thinning, or baldness. To avoid such undesirable effects, endoscopic-assisted harvest of the temporoparietal fascial flap was performed in 9 patients with microtia. Through two horizontal incisions in the temporal region, the temporoparietal fascia was dissected, and the flap was harvested using bipolar scissors and coagulating shears. Flaps were dissected and harvested successfully without any complications in 7 patients, although extra incisions were required to facilitate coagulation in 2 patients. The authors introduce this harvesting technique and describe some representative cases. Using endoscopic guidance, this is a versatile, safe procedure with minimal morbidity, and is applicable to other reconstructive procedures that require a temporoparietal fascial flap, including the free flap. PMID- 11037159 TI - Extraoral vs. intraoral distraction osteogenesis in the treatment of hemifacial microsomia. AB - During hemifacial microsomia (HMF), an important phase of the treatment is elongation of the hypoplastic mandible, mainly the ramus, at an early stage. Twenty-two patients with HFM were treated with distraction osteogenesis: 12 with an extraoral device (10 unidirectional and 2 multidirectional) and 10 with an intraoral device. The mean elongation with the extraoral device was 21 mm, and with the intraoral device was 17 mm, resulting in a more symmetrical facial appearance. The advantages and disadvantages of both methods are presented, based on the authors' experience and a review of the literature. The extraoral device permits elongation of a greater distance, enables extraoral control of the vector of elongation, and conserves the gonial angle by working in many directions. The main disadvantages of the extraoral device are the social inconvenience to the patient and the extraoral cutaneous scars. Conversely, the intraoral device is much more socially convenient to the patient and avoids residual scarring. However, in 2 patients treated with an intraoral device, an undesired contralateral open bite appeared as a result of reduced vector control. The intraoral method should always be considered first because of its previously mentioned advantages. However, in severely hypoplastic patients, when three dimensional correction and gonial angle control are necessary, or when there is a limited space along the planned distracted bone, the extraoral device has an advantage over the intraoral device. PMID- 11037160 TI - Reliability of island flaps raised after superficial and deep burn injury. AB - In select cases, to prevent any functional loss and to initiate early function during the early burn period, the reconstructive procedure of choice may be flap coverage. In these circumstances, when the ideal flap donor site is burned, the clinician may be hesitant to raise this flap because of questionable flap survival. The authors conducted this study to determine whether a superficially or deeply burned skin island flap would survive when elevated during the early postburn period. If these flaps are usable, they could expand the options available for burn wound coverage. They used a rat epigastric island flap model, and divided 50 study animals into two groups. In group 1 (N = 25), the right epigastric flap site was burned superficially and the left side was left uninjured. Island flaps were raised on both sides 4 days after the burn injury. The flaps were then sutured back into their original sites, and were evaluated 5 days after the surgery. In group 2 (N = 25), the right epigastric flap site was burned deeply and the left side was left uninjured. Island flaps were raised 4 days after the burn injury on both sides, as in group 1. The flaps were then sutured back into place and were evaluated 5 days after the surgery. All of the control flaps on the rats' left sides survived in both groups. In addition, all the superficially burned flaps survived in group 1 (100%), and 21 of the deeply burned flaps survived in group 2 (84%). There was no significant difference between superficially and deeply burned flaps with regard to survival, and the burned flaps were as successful as the unburned control flaps in both groups (p = 0.11). Skin island flaps elevated after superficial or deep burn injury are reliable in this animal model. PMID- 11037161 TI - The free musculocutaneous tensor fascia lata flap as a backup procedure in tumor surgery. AB - The musculocutaneous tensor fascia lata (TFL) flap provides a small muscle belly and a strong fascial layer in combination with abundant skin coverage (15 x 40 cm), which makes the flap an attractive unit for composite free tissue transfer. The free TFL flap was used in nine cases of recurrent cancer of the chest wall (N = 7) and the abdominal wall (N = 2). The mean size of the full-thickness defects after tumor excision measured 12 x 25 cm. The operating time ranged from 4 to 8 hours (mean operating time, 5.5 hours). The operation was performed with two teams, and no repositioning of the patient was necessary during the operation. By raising the TFL flap, no additional area of the trunk was involved. The authors did not experience a prolonged ventilation time in their group of multimorbid patients. The donor site was closed directly (4 of 9 patients) or split skin grafted (5 of 9 patients). There was no functional deficit. In one patient the venous anastomosis had to be revised. There were no further complications, and no flaps were lost. The hospital stay was short (21 days on average), the outcome successful, and primary healing was obtained. The free TFL flap proved to be a reliable flap that is easy technically to harvest. Thus the free TFL flap is a valuable backup procedure in tumor surgery. PMID- 11037162 TI - An anatomic examination of the suprascapular flap. AB - To investigate the potential utility of a suprascapular flap, the authors examined dissected cadavers to identify the ramification of the suprascapular artery, and directly injected 2.5% patent blue dye to observe the distribution of its cutaneous perforators. Results indicate that a suprascapular flap is not practicable. The branching pattern of the suprascapular artery from the subclavian artery varies widely, making dissection of the vessels highly problematic. Moreover, no constant presence of cutaneous perforators from the artery could be identified in either the suprascapular fossa or the shoulder. PMID- 11037163 TI - The effect of blood supply in muscle and an elevated muscle flap on endogenous tissue-engineered bone by rhBMP-2 in the rat. AB - Metabolism and remodeling of bony tissue are maintained and controlled by blood supply. In this study, bony tissue osteoinduced heterotopically by recombinant human bone morphogenetic protein-2 (rhBMP-2) in rat muscles with different amounts of blood supply was investigated. The implant beds were in the latissimus dorsi muscle (LDM) and in the space wrapped with an elevated latissimus dorsi muscle flap (LDMF). The blood flow was preestimated at two muscle sites using a laser Doppler blood flowmeter. The flow value of the LDM was 2.23 +/- 0.17 ml/min/100 g (mean +/- standard deviation). The flow values of the LDMF were 1.71 +/- 0.22 ml/min/100 g 30 minutes after elevating the LDMF, 1.75 +/- 0.20 ml/min/100 g after 1 week, 1.83 +/- 0.19 ml/min/100 g after 2 weeks, and 1.99 +/- 0.18 ml/min/100 g after 3 weeks. Bony tissues induced heterotopically by rhBMP-2 were examined radiographically and histologically. On radiographs, radiopaque shadows in the LDM were almost as large as those in the LDMF. The radiopacity in the LDM was a little higher than that of the LDMF. The microscopic findings showed increased trabeculae and more hematopoietic marrow in the LDM than lamellalike bone in the LDMF. Mean bony area in the implant was 1.05 +/- 0.26 mm2 in the LDM, and was 0.70 +/- 0.11 mm2 in the LDMF. Bony proportions in the overall implant area were 18.3 +/- 3.46% in the LDF and 12.1 +/- 2.18% in the LDMF. The current study indicates that blood supply is an important factor for promoting heterotopic osteoinduction by rhBMP-2 to produce massive bony tissue as an endogenous method of tissue engineering. PMID- 11037164 TI - Contribution of conventional axial computed tomography (nonhelical), in conjunction with panoramic tomography (zonography), in evaluating mandibular fractures. AB - Previous studies comparing the sensitivity between different radiological exams have concluded that conventional axial computed tomography (CT; nonhelical) is unsuitable in the assessment of mandibular fractures. Axial CT was shown to have a reduced sensitivity compared with plain radiographs and panoramic tomography because it missed nondisplaced fractures in the posterior portion of the mandible. Because the resolution of CT has improved from the time of these previous studies, the authors were interested in assessing whether axial CT (nonhelical) could now provide additional clinically useful information and enhance our understanding of mandibular fractures, beyond that obtained from panoramic tomography alone. In their study, 5 staff surgeons initially evaluated the panoramic tomograms and then the CT scans of 39 patients with 66 fractures. A series of four questions assessed the relative contribution of these two radiological exams in formulating an optimal operative plan for each patient. The authors found that axial CT provided supplementary information regarding missed fractures, comminution, and the exact size and degree of displacement of fracture fragments. This additional data could have changed the operative plan in a substantial proportion of patients (17 of 39). Axial CT demonstrated two missed parasymphyseal fractures (2 of 39 patients) that were not seen on these patients' panoramic tomograms. Axial CT also revealed undiscovered comminution or demonstrated fracture displacement more precisely in 39% of patients (15 of 39) and 24% of fractures (16 of 66). This study demonstrates that axial CT was clinically useful as an additional investigation to panoramic tomography. Axial CT helped elucidate further the nature of suspected mandibular fractures. PMID- 11037165 TI - A unique case of secondary microvascular glansplasty: the last genitoreconstructive frontier? AB - Isolated amputation of the noncarcinogenic penile glans is an extremely rare event. Immediate replantation as a composite graft or by microvascular techniques is the treatment of choice. Secondary reconstruction of the penile glans may be indicated after necrosis as a sequela of infection, surgical resection for malignancy, traumatic loss, and self-mutilation. To date, reported surgical techniques of glanular reconstruction have only been described as part of total phalloplasty. Secondary reconstruction of the penile glans using a sensate radial forearm free flap was performed in a 28-year-old man who presented 3 years after self-amputation in a psychotic state. PMID- 11037166 TI - Critical leg ischemia resulting from interruption of collaterals by harvest of the rectus abdominis free flap: endovascular salvage. AB - In the setting of aortoiliac occlusive disease, the inferior epigastric artery may be an important pathway for collateral blood supply to the lower extremities. A 72-year-old man developed critical ischemia of both legs after harvest of a rectus abdominis free flap as a result of interruption of the inferior epigastric artery. In patients with aortoiliac occlusive disease, the contribution of the inferior epigastric artery to lower extremity blood flow should be evaluated noninvasively. In the setting of reversed flow, the inferior epigastric artery usually should not be divided. PMID- 11037167 TI - Pregnancy as a tissue expander in the repair of a massive ventral hernia. AB - The use of tissue expanders has been described in numerous applications, including the closure of massive abdominal wall defects. The advantages of tissue expansion include providing adequate soft tissue for stable coverage of prosthetic material. In a subfascial plane, expanders can also expand muscle and fascia to allow total autologous repair of massive ventral hernias. These techniques for abdominal wall reconstruction are well established to cover viscera, to repair hernias, and to restore acceptable contour. The authors present the novel case of a woman with unstable skin graft coverage of a massive ventral hernia whose pregnancy was used as a surrogate intra-abdominal tissue expander to facilitate abdominal wall reconstruction. PMID- 11037168 TI - The sandwich technique for closure of a palatal fistula. AB - A full-thickness fistula of the hard palate can be closed by various methods. Recurrences are seen many times and more stable methods of closure have been researched. The authors attempted to close a palatal fistula by adhering to the main rule of reconstruction as stated by Gillies, "replace the lost tissues in kind." They used a buccinator musculomucosal transposition flap for the nasal lining, a cranial bone graft for the palatal bone, and a local mucoperiosteal transposition flap for the oral closure. The flaps and bone adapted well to the fistula. There were no recurrences during 12 months of follow-up. This "sandwich flap"--a three-layer closure--is a reliable technique for the repair of a full thickness palatal fistula. PMID- 11037170 TI - Giant plexiform neurofibroma of the back. AB - Complete excision of a giant neurofibroma can be technically difficult. Thorough preoperative planning with magnetic resonance imaging, computed tomography, and arteriography are indicated to define the extent of the mass and to facilitate operative planning. By following the treatment guidelines discussed in this case report, the authors feel that these tumors can be excised safely with minimal morbidity. PMID- 11037169 TI - Docetaxel (taxotere) extravasation: a report of five cases with treatment recommendations. AB - Docetaxel (Taxotere) is a relatively new antineoplastic agent that is proving to be clinically useful in the treatment of a number of major solid tumors, including breast, ovarian, lung, and prostate carcinoma. Common systemic toxicities include neutropenia, alopecia, nausea, and vomiting. The authors report 5 patients (age range, 54-89 years) who experienced extravasation injuries with dramatic clinical presentations during peripheral intravenous administration of docetaxel. The authors did not find any other reported series of docetaxel extravasation in an extensive literature review. They present these 5 patients and provide treatment recommendations. PMID- 11037171 TI - Multiple-digit onychomycosis: a simple surgical cure. AB - The management of intractable onychomycosis involving multiple fingernails continues to be optimized. Failure of pharmacological treatment necessitates operative intervention. Current surgical procedures are complex and have frequent donor site complications. This report describes a safe, simple, and effective surgical treatment that eradicates the disease, results in low morbidity, and yields high patient satisfaction. PMID- 11037172 TI - Unilateral virginal breast hypertrophy. AB - A case of unilateral virginal breast hypertrophy with a review of the etiological factors and treatment modalities is presented. A 16-year-old girl presented with progressive enlargement of the left breast of 5 months' duration. The result of the mammographic examination was consistent with cystosarcoma phyllodes. Fine needle aspiration biopsy revealed giant fibroadenoma. Although some of the characteristics of the fine-needle aspiration biopsy specimen were suspicious for cystosarcoma phyllodes, there were no adequate epithelial structures, which are obligatory for the diagnosis. The patient was treated with subcutaneous mastectomy and subpectoral insertion of a silicone gel implant. The histopathological examination was consistent with virginal hypertrophy. The breast maintained its volume with no further growth in the affected or in the normal breast after 4 years of follow-up. PMID- 11037173 TI - Sclerosing lobular hyperplasia of the breast after reduction mammaplasty. AB - Sclerosing lobular hyperplasia is a uncommon benign fibroproliferative lesion of the breast similar to a fibroadenoma. It usually presents as a mass in the outer quadrant of the breast of younger women. The authors report an atypical case of sclerosing lobular hyperplasia that presented as bilateral breast masses in an elderly woman after a reduction mammaplasty. Breast masses that develop after reduction are often a result of fat necrosis, internal scarring, or organizing hematomas. Those that do not resolve warrant open biopsy because they may represent malignant or benign neoplasms. PMID- 11037174 TI - Use of triamcinolone acetonide injection in ear reconstruction. AB - Reconstruction of the complicated contours of the ear is a difficult task. To maintain and recreate the reconstructed contours of ears masked by the following various reasons is even more difficult. During the first stage of cartilage transplantation, the ear contours are quite often masked as a result of prolonged edema; thick, draping skin; organized hematoma; or inflammatory exudates. Dressings and splints have been reported as management methods of the ear postoperatively after the first stage of cartilage transplantation, but it is difficult to use these for a period of 6 months, the time when edema subsides. The author used triamcinolone acetonide injection locally in the area of the scapha and triangular fossa of the reconstructed ear to recreate the contours that were masked. This is a simple and effective method for achieving the desirable contours. PMID- 11037175 TI - The "stegosaurus" dressing: a simple and effective method of securing skin grafts in the burn patient. AB - Skin grafts are vulnerable to shear stress, infection, and hematoma formation during the postoperative period, all of which reduce graft survival. Various methods of dressing application and materials have been described in the literature to try and prevent graft loss. The authors report the use of the "stegosaurus dressing" (Eggcrate Pad) in 6 patients to secure skin grafts. Patients chosen were those who were either noncompliant or who sustained burns in unfavorable anatomic sites. All grafts demonstrated complete take without infection and hematoma formation. This foam dressing provides an even pressure to the recipient bed, absorbs drainage, and protects the graft from shearing. It also demonstrates the versatility to be used in difficult nonburn skin graft areas. The stegosaurus dressing is easy to apply, inexpensive, and provides a very secure dressing over the skin graft. PMID- 11037176 TI - Endoscopic harvest of pectoralis major muscle flap for correction of a neck deformity. PMID- 11037177 TI - Drains in reduction mammaplasty. PMID- 11037178 TI - Rhomboid flap for surgery of the scalp with or without expansion: an experimental study. PMID- 11037179 TI - History of neuropathology in Japan. PMID- 11037180 TI - Encephalomyelitis, brain tumors, neuromuscular diseases and miscellaneous disorders. AB - Japanese neuropathologists have accomplished and contributed to a considerable number of achievements, and some of these are cited in other articles in this issue. Several of these achievements as well as other miscellaneous discoveries are briefly summarized in the present paper. Specifically these relate to rabies postvaccinal encephalomyelitis, experimental allergic encephalomyelitis, brain tumor research, neuromuscular disorders, schizophrenia, viral infections, glial inclusion body in multiple system atrophy, and the neurobiology of glia. PMID- 11037181 TI - Minamata disease. AB - Minamata disease (methylmercury poisoning) was first discovered in 1956 around Minamata Bay, Kumamoto Prefecture, Japan. A second epidemic in Japan occurred in 1965 along the Agano River, Niigata Prefecture. This paper presents a brief review of Minamata disease with an emphasis on the cases found in Kumamoto Prefecture. At autopsy, the most conspicuous destructive lesion in the cerebrum was found in the anterior portions of the calcarine cortex. Less severe but similar lesions may be found in the post-central, pre-central and temporal transverse cortices. Secondary degeneration from primary lesions may be seen in cases with long survival. In the cerebellum, pathological changes occur deep in the hemisphere. The granule cell population was more affected, compared with Purkinje cells. Among peripheral nerves, sensory nerves were more affected than motor nerves. Our recent experimental studies that reveal knowledge of the pathogenesis of methylmercury poisoning will be discussed. PMID- 11037182 TI - Subacute myelo-optico-neuropathy: clioquinol intoxication in humans and animals. AB - It remains a tragic event that some 10,000 individuals in Japan developed a unique neurologic disease, subacute myelo-optico-neuropathy (SMON). Many of the affected patients still suffer serious sequelae, such as dysesthesia and muscle weakness in the lower extremities, and loss or deficits in visual acuity. Neuropathologic studies on SMON patients and experimental reproduction of the disease in animals which had been administered clioquinol helped resolve the etiology of this disease. Common pathologic features seen in SMON patients and in dogs and cats chronically intoxicated with clioquinol were distal dominant axonopathy, mainly in the spinal long tracts and optic tracts. Particular abdominal symptoms present in patients after clioquinol ingestion could also be reproduced experimentally in dogs. SMON research in Japan may be worth reviewing for determining the etiology and preventing similar neurotoxic diseases in the future. PMID- 11037183 TI - Nasu-Hakola disease: a review of its leukoencephalopathic and membranolipodystrophic features. AB - The clinicopathological features of Nasu-Hakola disease are described by reviewing previously reported cases and adding consideration to newly disclosed evidence. This disease is an autosomal recessive disorder characterized by membranocystic lipodystrophy in the skeletal system and sclerosing leukoencephalopathy in the nervous system. The leukoencephalopathic alterations are demyelinization of the cerebral white matter, associated with conspicuous fibrillary gliosis and preservation of the subcortical arcuate fibers. Sudanophilic granules are focally scattered in the perivascular space or widely infiltrated in the affected white matter, and some neuronal loss with deposits of calcospherites is encountered in the basal ganglia and also in the thalamus. Spheroid formation with an increased number of neurofilaments in the neuronal axon is considered a possible pathogenesis, and a primary vascular mechanism is also suggested. Interestingly, most of the reported cases of Nasu-Hakola disease are from Japan and Finland which suggests heredofamilial background as a cause. PMID- 11037184 TI - Neuropathological findings of Onuf's nucleus and its significance. AB - Onuf's nucleus is a small group of cells which are located mainly in the anterior horn of the second sacral segment of the spinal cord. This paper describes the history of studies relating to this nucleus including a discussion of its relation with the various pathological studies which have been made. PMID- 11037185 TI - Muscular dystrophy. AB - Muscular dystrophy is a group of genetically determined muscular disorders marked by progressive wasting and weakness of the skeletal muscle, but which often affect cardiac and smooth muscles or other tissues. The patterns of inheritance are either dominant or recessive although the gene may be defective because of a new mutation. Growing evidence revealed the marked heterogeneity of the muscle disorders, and considerable numbers of Japanese scientists and physicians have contributed to the research progress in muscular dystrophy. Among these the discovery of an increased serum creatine kinase activity in muscular dystrophy opened the way for the most reliable laboratory test for muscular dystrophy in 1959, and subsequently accelerated progress in a broad range of research areas in medicine. Progress in modern genetics and molecular pathology provided another breakthrough in muscular dystrophy research and, in 1987, dystrophin was identified, a deficiency of which causes DMD. The present review article highlights contributions of Japanese scientists to muscular dystrophy research. PMID- 11037186 TI - Hereditary dentatorubral-pallidoluysian atrophy. AB - Pathology and associated clinical symptoms of hereditary dentatorubral pallidoluysian atrophy (H-DRPLA) which was established as a new inherited neurodegenerative disease in 1982 are described. Obligatory lesions in the central nervous system combine with degeneration of the dentatorubral and pallidoluysian pathway, and occasional degenerative lesions are found in the cerebral white matter, putamen, Goll's nucleus of the medulla oblongata, and lateral corticospinal and Goll's tract of the spinal cord. The main clinical symptoms are myoclonus, epilepsy, dementia or mental retardation, cerebellar ataxia and choreoathetosis. Furthermore, newly developing aspects in the pathology of H-DRPLA following the discovery of the gene locus of H-DRPLA in 1994 are briefly described. PMID- 11037187 TI - Pathology of familial amyloidotic polyneuropathy with TTR met 30 in Kumamoto, Japan. AB - Seventeen autopsy and 12 sural nerve biopsy cases of familial amyloidotic polyneuropathy (FAP) with transthyretin (TTR) Met 30 were examined clinicopathologically. In the autopsy cases, amyloid deposits were prominent in the peripheral nerve tissues, autonomic nervous system, choroid plexus, cardiovascular system and kidneys. Amyloid involvement in the posterior and anterior roots of the spinal cord, spinal ganglia, thyroid and gastrointestinal tract were also frequent. In the sural nerve biopsy, degenerative changes corresponding in degree to the duration of the clinical course were observed in the endoneurium, and amyloid deposition occurred around the blood vessels. Electron microscopy revealed degenerative changes in the axon, myelin sheath and Schwann cells. The morphometric study showed decreased numbers of small-caliber myelinated fibers during the early stage. TTR was confirmed immunohistochemically as a major component of amyloid deposits. In transgenic mice carrying the human mutant TTR gene, amyloid deposition was observed in various organs except in the peripheral nerves and choroid plexus. Liver transplantation therapy to FAP patients has been carried out and future follow-up studies should investigate the effects of therapy. PMID- 11037188 TI - Creutzfeldt-Jakob disease. AB - It is valuable to summarize the milestone study of prion diseases done in Japan for review in the journal Neuropathology in 2000. Many studies done in Japan promote world prion research activity, and also influence further research projects in other groups abroad. In this review the author focuses on the transmission experiment, the discovery of abnormal prion protein localization in the synaptic structures or follicular dendritic cells, and the genetic analysis of prion protein gene for the establishment of familial prion diseases. PMID- 11037189 TI - Untangling Alzheimer's disease from fibrous lesions of neurofibrillary tangles and senile plaques. AB - Neuropathological evidence suggests that the two fibril lesions of neurofibrillary tangles (NFT) and senile plaques are the major findings in brain tissue of Alzheimer's disease (AD) and that their occurrence is strongly associated with the symptoms of dementia. Genetic findings have indicated that the pathological molecules from the lesions function as causal agents. There is little evidence, however, to directly indicate that fibril lesions themselves kill neuronal cells in vivo. In spite of such limitations it is important to consider the molecular events involved in AD etiology. In this review of the contribution of Japanese neuropathologists to studies of AD, I will introduce briefly their work and highlight some current topics for consideration on the etiology of AD, and the basis of cell death, and will offer my perspective on outstanding conflicting issues. PMID- 11037190 TI - Moyamoya disease. AB - Moyamoya disease is a specific chronic cerebrovascular occlusive disease first reported by Japanese surgeons in 1957. The disease is characterized by stenosis or occlusion of the terminal portions of the bilateral internal carotid arteries and abnormal vascular network in the vicinity of the arterial occlusion. It may cause ischemic attacks or cerebral infarction, which is more frequent in children than in adults. In adults, cerebral hemorrhage may occur. The disease is distributed in all age groups, but the highest peak is in childhood at less than 10 years of age. The characteristic histopathologic features of the steno occlusive arteries are fibrocellular thickening of the intima containing proliferated smooth muscle cells and prominently tortuous and often duplicated internal elastic lamina. There is usually no atheromatous plaque in the arterial wall. Etiology of the disease is still unknown; however, multifactorial inheritance is considered possible because of a higher incidence of the disease in Japanese and Koreans and approximately 10% of familial occurrence among the Japanese. Recent genetic studies suggest some responsible genetic foci in chromosomes 3, 6 and 17. PMID- 11037191 TI - HTLV-I-associated myelopathy. AB - HTLV-I was first described as a pathogenic human retrovirus that causes adult T cell leukemia (ATL). Soon after the discovery of HTLV-I, an association of this virus with a slowly progressive neurological disorder was found independently in Japan and Caribbean islands, and this new clinical entity (HTLV-I-associated myelopathy with tropical spastic paraparesis) was named HAM/TSP. Autopsy findings clarified the chronic inflammatory nature of the disease. Detailed neuropathological analysis demonstrated: (i) T-cell-dominant mononuclear cell infiltration; (ii) diffuse and symmetrical degeneration of the anterolateral and inner portion of the posterior columns involving both myelin and axons; (iii) the presence of cytotoxic T cells and apoptosis of helper/inducer T cells; (iv) in vivo localization of HTLV-I provirus in the perivascular infiltrated T cells; and (v) accentuation of inflammatory lesions at the site with slow blood flow. From these findings it is suggested that a T-cell-mediated chronic inflammatory process targeting the HTLV-I-infected T cells is the primary pathogenic mechanism of HAM/TSP. PMID- 11037192 TI - Crow-Fukase syndrome. AB - Crow-Fukase syndrome is a unique multisystem disorder that is also known as POEMS syndrome (an acronym for polyneuropathy, organomegaly, endocrinopathy, the presence of M-protein and skin change). This syndrome is strongly associated with plasma cell dyscrasia. Circulating light chains of M component, almost invariably IgG lambda or IgA lambda, are found in 75% of patients. Neuropathologically, segmental demyelination, particularly in the proximal segment of the peripheral nerve trunk, is the primary process. Axonal degeneration and marked endoneurial edema are also characteristic. Focal excessive myelin outfolds with globular features corresponding to periodicity and paranodal enlargement of myelin are also highly characteristic of this syndrome. Vascular endothelial growth factor (VEGF) was found to be specifically and highly elevated in the serum of patients with this syndrome, suggesting a pathogenic role. M-protein, interleukin (IL) 1beta, IL-6 and tumor necrosis factor (TNF)-alpha are also considered to be involved in the pathogenesis. Treatment consists of radiation and surgical resection of the myeloma, chemotherapy, and a high dose of intravenous immunoglobulin (IVIg). PMID- 11037193 TI - Diffuse Lewy body disease. AB - Diffuse Lewy body disease (DLBD) has been studied from various viewpoints and, although clinical diagnostic criteria for DLBD have been proposed, diagnosis remains difficult. DLBD has been reported to be the second most common form of dementia in the aged, following Alzheimer-type dementia. It has, however, been clinically under-diagnosed. Therefore, the search for diagnostic markers for DLBD must continue. Very recently, 'dementia with Lewy bodies' (DLB) was proposed as a generic term for various forms of dementia with Lewy bodies, including DLBD and similar disorders. Cortical Lewy bodies are the most important pathologic marker for diagnosis of DLBD. At present, however, the mechanism responsible for cortical Lewy body formation has yet to be disclosed. PMID- 11037194 TI - Dementia with motor neuron disease. AB - Dementia with motor neuron disease has been described as a new clinicopathologic entity and more than 100 cases have been reported in Japan since 1964. The clinicopathologic criteria in the diagnosis of dementia with motor neuron disease are: (i) frontotemporal lobe-type dementia with insidious onset, mostly in the presenile period; (ii) neurogenic muscular wasting during the course of the illness (amyotrophic lateral sclerosis- or SPMA-like symptoms); (iii) duration from the onset of illness to death of 2-5 years (average, 30.6 months); (iv) both extrapyramidal symptoms and definite sensory deficits are present less commonly; (v) no characteristic abnormalities in the cerebrospinal fluid or electroencephalogram on screening; (vi) no known parental consanguinity or familial occurrence; and (vii) non-specific, mild to slight degenerative changes in the frontotemporal cortex, hypoglossal nuclei and spinal cord, and frequently in the substantia nigra. Dementia with motor neuron disease is characterized by ubiquitin-immunoreactive intraneuronal inclusions in cortical layer II and hippocampal dentate granule cells. PMID- 11037195 TI - Mitochondrial encephalomyopathy. AB - Mitochondrial encephalomyopathy is a disease based on multisystemic mitochondrial dysfunction. Pathologic, biochemical and molecular genetic approaches to the disease have revealed the complex features of the phenotype and its relationship to the genotype. Last decade's great success of the mtDNA study must move research of the disease into the next phase. PMID- 11037196 TI - Autosomal recessive juvenile parkinsonism: a key to understanding nigral degeneration in sporadic Parkinson's disease. AB - The contribution of genetic factors to the pathogenesis of Parkinson's disease (PD) is supported by the demonstration of the high concordance in twins studies using positron emission tomography (PET), the increased risk among relatives of PD patients in case-control and family studies, and the existence of familial PD and parkinsonism by single gene defect. Recently several genes have been mapped and/or identified. Alpha-synuclein is involved in a rare dominant form of familial PD with dopa-responsive parkinsonism features and Lewy body-positive pathology. In contrast, parkin is responsible for the autosomal recessive form (AR-JP) of early onset PD with Lewy body-negative pathology. The clinical features of this form include early onset (in the 20s), levodopa-responsive parkinsonism, diurnal fluctuation, and slow progression of the disease. Parkin consists of 12 exons and the estimated size is over 1.5 Mb. To date, variable mutations such as deletions or point mutations resulting in missense and nonsense changes have been reported in AR-JP patients. In addition, the localization of parkin indicates that parkin may be involved in the axonal transport system. More recently we have found that parkin interacts with the ubiquitin-conjugating enzyme E2 and is functionally linked to the Ub-proteasome pathway as a ubiquitin ligase, E3. These findings fit the characteristics of a lack of Lewy bodies (these are cytoplasmic inclusions that are considered to be a pathological hallmark). Our findings should enhance the exploration of the mechanisms of neuronal death in PD as well as other neurodegenerative disorders of which variable inclusion bodies are observed. PMID- 11037197 TI - Juvenile muscular atrophy of distal upper extremity (Hirayama disease): focal cervical ischemic poliomyelopathy. AB - Hirayama disease is characterized by an initially progressive muscular weakness and atrophy of the distal upper limb(s) in adolescence, occurring predominantly in males, followed by spontaneous arrest within several years. Although the disease could be separated from motor neuron disease, some authors considered the illness to be a variant of degenerative motor neuron disease until the first autopsy case was reported which showed focal ischemic changes in the anterior horn of the lower cervical cord. Since then, many in Japan have recognized the disease as cervical ischemic poliomyelopathy; however, several authors in foreign countries did not or do not appreciate the pathologic findings of the disease, and still consider the illness a degenerative motor neuron disease. The pathology of the disease prompted neuroradiologic investigations, which have revealed dynamic changes of the cervical dural sac and spinal cord induced by neck flexion. The cause of these dynamic changes is unknown. However, as the number of patients is exceedingly large in Japan, there may be an ethnic factor. PMID- 11037198 TI - Delayed neuronal death. AB - A brief, transient episode of cerebral ischemia causes selective loss of CA1 pyramidal cells in the hippocampus. Interestingly, it takes 2-3 days for the neuronal damage of CA1 to become morphologically obvious; hence, the term 'delayed neuronal death' was coined to describe this phenomenon. Approximately two decades after it was first recognized, the precise mechanism of delayed neuronal death is not yet fully understood. The most widely supported explanation is the glutamate-Ca2+ theory, in which vulnerability of CA1 neurons is ascribed to their property of neurotransmitters and their receptors. Although this theory explains the early phase of neuronal response to ischemia fairly well, it does not offer a satisfactory explanation for the delayed phase of the process. Excessive accumulation of Ca2+ is observed in CA1 following transient ischemia, but accumulating evidence suggests that additional changes at the molecular level have to follow to completing the dying process, including expression of various apoptosis-related genes. Further studies are, therefore, necessary to elucidate the mechanism of delayed neuronal death which, in turn, may open a new window for the treatment of stroke. PMID- 11037199 TI - Visiting scientists to the USA from Japan and their representative works. PMID- 11037200 TI - 100 years of collaboration between Japanese and German neuropathologists. PMID- 11037201 TI - France and Japan in neuropathology. PMID- 11037202 TI - International exchange program. UK and Japan. PMID- 11037203 TI - Neuropathological relationships between Austria and Japan. PMID- 11037204 TI - International exchange program. Canada and Japan. PMID- 11037205 TI - Medical sociology at the start of the new millennium. PMID- 11037206 TI - Residential segregation and the epidemiology of infectious diseases. AB - Several empirical studies have documented the effects of residential segregation on health inequalities between the US African-American and white populations. However, the majority of such studies have not explained the pathways that link residential segregation and specific health outcomes. This paper presents a conceptual framework of the role that residential segregation may play in the epidemiology of tuberculosis (TB) and other infectious diseases. This is an important issue given the concentration of TB cases among US racial/ethnic minorities and the increasing gap in the incidence of infectious diseases between minorities and the white majority. Segregation may have an indirect effect on the transmission of TB because of its negative impact on the quality of neighborhood environment in segregated communities. Segregation concentrates poverty, overcrowded and dilapidated housing and social disintegration in minority areas, and results in limited access to health care. Furthermore, two dimensions of residential segregation (isolation and concentration) may have direct effects on TB transmission. The isolation of minorities confines TB to segregated areas and prevents transmission to the rest of the population. High-density levels in minority areas increase the probability of transmission within the segregated group. In order to operationalize the above pathways, health researchers may rely on the segregation literature, which has conceptualized various dimensions of residential segregation and proposed ways to measure them. The indirect pathways that link segregation and TB can be captured through exposure indices, which quantify the concentration of risk factors for TB for various racial and ethnic groups. The direct pathways can be captured through the isolation index (which is a proxy for the degree of interaction between the segregated group and the rest of the population) and two proposed measures of density (which are proxies for the likelihood of transmission within the segregated group and from the segregated group to the rest of the population). PMID- 11037207 TI - Disability, space and sexuality: access to family planning services. AB - In this paper we examine, from a social perspective, access to family planning clinics for disabled people. We argue that disabled people are commonly understood to be either asexual, uninterested in sex or unable to take part in sexual activity, or sexual 'monsters' unable to control their sexual drives and feelings. These understandings are reproduced through the use of cultural representations and myths, and are evidenced in the planning and design of family planning clinics and the information and services they provide. To illustrate our arguments we present the findings of a short questionnaire survey of all family planning clinics in Northern Ireland. Physical access to these clinics was partial, and access to information and services were extremely limited. These results indicate that disabled people are not expected to be using the services (consultation, treatment, information) that family planning clinics provide. As such, family planning clinics in Northern Ireland represent a landscape of exclusion, denying disabled people access to services and reproducing cultural ideologies concerning disability and sexuality. PMID- 11037208 TI - Mortality differentials among women: the Israel Longitudinal Mortality Study. AB - The first aim of this study was to examine differentials in mortality among Israeli adult women with respect to ethnic origin, marital status, number of children and several measures of socio-economic status; the second was to compare mortality differentials among women with those found for Israeli men. Data are based on a linkage of records from a 20% sample of the 1983 census with the records of deaths occurring until the end of 1992. The study population includes 79,623 women and the number of deaths was 14,332. Measures of SES included education, number of rooms, household amenities and possession of a car. Results indicated higher mortality among women originating from North Africa compared with Asian and European women. Adjustment to SES eliminated the excess mortality among North African women and revealed a lower mortality of Asian women, relative to Europeans. Among women aged 45-69, substantial and consistent mortality differentials were evident for all SES indicators examined where mortality declined with improved socio-economic position. Mortality was related to women's childbearing history, with the highest mortality among childless women. Mortality differentials among women aged 70+ were generally narrower than those found for younger women. Gender differences in mortality differentials varied by the socio demographic indicator and age. PMID- 11037209 TI - Stage of change is associated with assessment of the health risks of maternal smoking among pregnant women. AB - This study explored pregnant women's assessment of the health risks associated with maternal smoking. The aim was to determine if stage of change relating to smoking is associated with risk assessment. A cross-sectional survey (employing a self-completion questionnaire) was conducted of all women who attended antenatal clinics at Leicester Royal Infirmary, National Health Service Trust, UK over a 2 week period. Questionnaires were completed by 254 respondents. Twenty seven percent of non-smokers agreed with more than 75% of a series of statements about the dangers of maternal smoking compared to 5% of smokers and 44% of women in social class I (highest social class) agreed with more than 75% of the statements compared with only 10% of women in social classes IV and V (lower social class groups). Married women were twice as likely to concur with more than 75% of the health risks compared to single or cohabiting women and 29% of women intending to breastfeed agreed with more than 75% of the statements compared with only 8.7% of women not intending to breastfeed. There was no significant effect of age, whether the pregnancy was planned, previous obstetric complications or whether the woman had a child with asthma or respiratory infections. A multiple regression analysis indicated that smokers were much less likely to agree with the health risks than their non-smoking counterparts (p = 0.034). Stage of change was related to both the number of health risks agreed with and the level of conviction. A woman's stage of change could be assessed at the start of antenatal care so that appropriate smoking cessation advice can be offered. PMID- 11037210 TI - Driving donation: a geographic analysis of potential organ donors in the state of Ohio, USA. AB - For many years, researchers of organ transplantation have primarily studied the differences in ethnic, cultural and religious characteristics of organ donors across the United States. Unfortunately, this work has failed to include a spatial parameter. Spatial analysis can play an important role in helping researchers better understand these characteristics across diverse landscapes. Using basic statistical techniques, a geographic information system (GIS) and data acquired from actual drivers license records, this study examines the demographic, socioeconomic, religious and electoral landscapes as they relate to potential organ donorship rates across the state of Ohio. Results indicate that income and education are important factors, and political affiliation, race and proximity to organ procurement organizations can also affect one's potential to donate. PMID- 11037211 TI - To boil or not: drinking water for children in a periurban barrio. AB - Boiling water, or other water purification methods, are common recommendations of health promoters in developing countries to improve the quality of drinking water in an attempt to decrease the incidence of childhood diarrhea. Health education programs frequently employ an approach based on knowledge deficits to promote this practice. However, there has been little published about water purification practices or associated variables such as knowledge deficits. We interviewed 266 randomly selected child caregivers about water purification in a poor periurban district of Santo Domingo, Dominican Republic. Though most reported that they provided purified drinking water for their children when they were babies, only half of the children five years of age and under were regularly drinking purified water at the time of the study. Only one knowledge variable remained significantly related to purifying drinking water in the final multivariate model. Other factors that remained in the final model were level of maternal education, endorsing being too tired to boil water and a global measure of social support. Several other hypothesized variables were not related to purifying water. Knowledge deficits may play only a limited role in determining this prevention practice. Further work is required to better identify key factors to improve this practice and hence guide health promotion efforts. PMID- 11037212 TI - Contract medicine arrangements in Hong Kong: an example of risk-bearing provider networks in an unregulated environment. AB - It is increasingly common in Hong Kong and elsewhere for employers to contract directly with physician networks to provide medical services to employees. These contracts are known in Hong Kong as contract medicine arrangements. In other countries and areas, managed care organizations are generally required by regulation or legislation to ensure that services of adequate quality are provided to patients who are locked in to network providers. There are no such requirements in Hong Kong and concerns have been raised about potential quality and cost trade-offs in contract medicine arrangements. Satisfaction surveys were sent to contract medicine enrollees in one large company in Hong Kong. The response rate was 30% and analysis of non-respondent data shows that respondents were representative of their group. Comparison of satisfaction using logistic regression showed that risk-bearing networks paid by capitation had consistently lower satisfaction ratings across all major dimensions including access, interpersonal care, communication with the doctor, choice of doctor, and outcomes. These findings suggest that quality, at least as perceived by the patient, may be lower in these networks. The issue is of concern in Asia where infrastructures and data systems are not well developed to adequately monitor quality of care or protect patient interests. This study highlights the need to structure pre-paid provider networks and managed care organizations so that quality of care is not compromised. At a time when managed care concepts are being applied throughout Asia, we believe attention needs to be drawn to this problem. PMID- 11037213 TI - "To every thing there is a season"--social time and clock time in addiction treatment. AB - Research on therapeutic interventions and the development or efficacy of treatment services consider 'time' only as a technical, 'objective' condition. Time series analysis and cohort studies describe changes in addiction careers over time, but fail to take into account the role of 'subjective' or 'social time', e.g. the functions of organisational and individual patterns of time use and time budgets. This paper reviews the notion of 'time' in addiction treatment systems. More specifically, the explicit or implicit role played by 'the time factor' in specific types of treatment such as '12-step programs', in-patient, out-patient care and individual treatment plans differs considerably and implies a re-definition and interpretation of 'Past', 'Present' and 'Future'. Temporal conceptions and time estimations of patients and therapists may influence the access to treatment and treatment outcomes. Societal values--lack of time in affluent societies--and a general acceleration in the fields of communication, consumption, work and leisure are mirrored in the treatment system. Recovery as a long-lasting learning process stands in sharp contrast to the 'quick fix'. The question is raised whether a post-modern concept of time is gaining importance as a counter movement, promoting a more individualised and differentiated treatment response and not any longer based on assumingly objective, technical criteria such as cost-efficiency. More research is needed on group-specific time concepts in treatment programs and the acceleration hypothesis in treatment systems. PMID- 11037214 TI - Negotiating natural death in intensive care. AB - Recent empirical evidence of barriers to palliative care in acute hospital settings shows that dying patients may receive invasive medical treatments immediately before death, in spite of evidence of their poor prognosis being available to clinicians. The difficulties of ascertaining treatment preferences, predicting the trajectory of dying in critically ill people, and assessing the degree to which further interventions are futile are well documented. Further, enduring ethical complexities attending end of life care mean that the process of withdrawing or withholding medical care is associated with significant problems for clinical staff. Specific difficulties attend the legitimation of treatment withdrawal, the perceived differences between 'killing' and 'letting die' and the cultural constraints which attend the orchestration of 'natural' death in situations where human agency is often required before death can follow dying. This paper draws on ethnographic research to examine the way in which these problems are resolved during medical work within intensive care. Building on insights from the literature, an analysis of observational case study data is presented which suggests that the negotiation of natural death in intensive care hinges upon four strategies. These, which form a framework with which to interpret social interaction between physicians during end of life decision making in intensive care, are as follows: firstly, the establishment of a 'technical' definition of dying--informed by results of investigations and monitoring equipment--over and above 'bodily' dying informed by clinical experience. Secondly, the alignment of the trajectories of technical and bodily dying to ensure that the events of non-treatment have no perceived causative link to death. Thirdly, the balancing of medical action with non-action, allowing a diffusion of responsibility for death to the patient's body; and lastly, the incorporation of patient's companions and nursing staff into the decision-making process. PMID- 11037215 TI - Neutralizing differences: producing neutral doctors for (almost) neutral patients. AB - Today 50% of medical students in Canada are women; they come from a wide range of racial, cultural, academic, and class backgrounds; they may openly identify as gay or lesbian. Yet to the extent that professional socialization produces uniformity of values, attitudes and future practice styles, the impact of increasing diversity is lessened. Based on a survey with undergraduate medical students, interviews with 25 students, and interviews with 23 faculty members and administrators at one Canadian medical school, this paper argues that there are impetuses within believe that the social class, 'race', ethnicity, gender, sexual orientation of a physician is not--and should not be--relevant during physician patient interactions. In short, intentional and unintentional homogenizing influences in their training work to neutralize the impact of increasing social differences among medical students. PMID- 11037216 TI - The co-occurrence of correct and incorrect HIV transmission knowledge and perceived risk for HIV among women of childbearing age in El Salvador. AB - This article examines the co-occurrence of correct and incorrect knowledge about documented and undocumented modes of HIV transmission among women of childbearing age in El Salvador, and the relationship between HIV transmission knowledge and perceived risk. Incorrect beliefs about HIV transmission co-occur at high levels with, and are largely independent of, accurate knowledge about documented modes of transmission. The co-occurrence of correct and incorrect HIV transmission knowledge was shown to have important implications for perceived risk. Both correct and incorrect HIV transmission knowledge increased the odds of risk perception; uncertainty about risk was decreased among those with higher levels of correct knowledge and increased among those with higher levels of incorrect knowledge. Among those who considered themselves to be at some risk for HIV, higher levels of correct knowledge reduced uncertainty about the degree of risk, while higher levels of incorrect knowledge increased the degree of risk perceived. High levels of endorsement of the documented modes of HIV transmission do not necessarily indicate accurate or adequate knowledge about HIV transmission in the population. Co-occurring inaccurate beliefs about undocumented modes of transmission reflect cultural understandings of contagion and disease, and influence how individuals make sense of medical-scientific information about transmission. Our results suggest that the co-occurrence of correct and incorrect HIV transmission knowledge shapes individual-level risk perceptions. Given the independence of accurate knowledge and inaccurate beliefs. HIV/AIDS education and prevention programs must seek to directly undermine inaccurate beliefs about HIV transmission as part of their efforts to promote behavior change. PMID- 11037217 TI - Place, social exchange and health: proposed sociological framework. AB - A sociological framework is proposed to better understand how spatial characteristics translate into people's physical and psychosocial conditions that are relevant to their health. In particular, high susceptibility to poor health among specific adult population groups is analyzed in terms of exclusion from or inadequate participation in a society's structure of opportunities. Acquisition of, and agency through, core social roles, such as the work role, the family and marital role, and civic roles, are essential prerequisites for successful personal self-regulation in adult life, strengthening a sense of self-esteem, self-efficacy, and belonging (self-integration). It is argued that exclusion from, or loss of core social roles, threats to their continuity and confinement to non-reciprocal exchange impair personal self-regulation and trigger a state of 'social reward deficiency'. This state, in turn, elicits prolonged stressful experience, and it may reinforce a person's craving for stress-relieving, potentially addictive health-damaging behavior. This framework is applied to the explanation of the life expectancy gap between Western and Central/Eastern European countries. Although most of the epidemiological evidence reviewed in support of this approach originates from investigations that were conducted in western countries several results reported in the collection of articles published in this Special Issue of Social Science and Medicine are in line with the proposed framework. PMID- 11037218 TI - A multilevel city health profile of Moscow. AB - This report describes a multilevel city-wide profile of physical health in Moscow, examining individual and urban level factors. Objectives of the paper were to: (1) identify macro and micro risk factors for poor physical health in Moscow;(2) assess the effect of two dimensions of micro determinants--personal health habits and social connectivity, such as social cohesion, social support, and social networks; (3) examine the hypothesis that relative social inequality is a significant structural condition at the community level which influences the physical health of individuals, as a main and as a joint effect with psychosocial behaviors. A random sample of Moscow adults, with household telephones (N = 2000), was collected 17-19 September, 1991, and had a completed interview rate of 81.8%. The questionnaire replicated items from the California Alameda Study and the US Health Interview Survey. Respondents' urban area of residence was linked to macro measures of inequality derived from the Moscow census. This report describes the baseline survey of a prospective study design. Results of this study demonstrate that the social context in a community affects the health of people living there independently from the effects of individual health lifestyle or social connectivity. The structural conditions in Moscow which significantly increased the vulnerability of specific social groups for poor physical health were identified in a hierarchical linear regression: relative social inequality in the form of income inequality; urban area poverty risks; and mean level of alcohol consumption in urban areas. The psychosocial conditions included poor diet, lack of social cohesion and social support, involvement in formal social networks in the form of professional groups. A multilevel theoretical perspective is important for defining the targets of preventive health policy by identifying the structural conditions which increase the health disadvantage of some social groups. Further research is needed in refining the concept of relative social inequality, as well as investigating whether psychosocial factors, such as social cohesion, are mediating links between sick societies and their sick citizens. PMID- 11037219 TI - Health lifestyles in Russia. AB - Utilizing data from the nationwide Russian Longitudinal Monitoring Survey (RLMS), this paper analyzes health lifestyles in Russia. Heavy alcohol use and smoking, a high-fat diet, and lack of leisure-time exercise are the principal culprits in fostering high rates of heart disease and other causes of premature mortality. This is especially the situation for middle-age, working-class males whose mortality is greater than any other segment of society. This paper focuses on alcohol use as a particularly lethal component of male lifestyles and presents a theoretical argument, grounded in the work of Weber and especially Bourdieu, that poor health lifestyles practices are largely the outcome of structural conditions (life chances) rather than agency (life choices). Societal and group norms and routine practices can adversely affect longevity and this is the case for Russian male blue-collar workers in the middle period of their life course. PMID- 11037220 TI - Health-related lifestyles and alienation in Moscow and Helsinki. AB - Health-related lifestyles (smoking, drinking alcohol, exercise and diet) and feelings of alienation (powerlessness and hopelessness) of the citizens of Helsinki and Moscow are examined and discussed in a framework of life chances and life choices. The data were collected by a postal survey of 18-64 yr old citizens of Helsinki (N = 824) and Moscow (N = 545) in 1991. Almost all respondents in both cities used alcohol, but heavy drinking was more frequently reported in Helsinki. Muscovite men were smokers more often and Muscovite women less often than their counterparts in Helsinki. Nearly half of the Muscovites, but less than one-fifth of the Helsinki respondents considered their diet unhealthy or of poor quality. Regular exercise was much more common among the Finns compared to the Muscovites. The sex difference in health-related lifestyles was wider in Moscow than in Helsinki, especially concerning health-damaging behaviour. Feelings of alienation were more pronounced in Moscow. In both cities alienation was more clearly associated with socioeconomic life chance factors than with lifestyle factors. In Helsinki feelings of alienation had stronger associations both with health and health related lifestyles, which possibly points to a conventional stratification effect of a market-based class society. In Moscow, which represents a more traditional community, alienation seemed to be part of a widely felt general discontent. Health was a highly salient value in both cities, especially among women. In Helsinki a high valuation of health was connected with less smoking, more exercise and a healthier diet. Valuing health did not seem to emerge as a distinct healthy lifestyle in Moscow where behavioural choices were limited by many material constraints. PMID- 11037221 TI - Socioeconomic factors, material inequalities, and perceived control in self-rated health: cross-sectional data from seven post-communist countries. AB - This study examined the association between perceived control and several socioeconomic variables and self-rated health in seven post-communist countries (Russia, Estonia, Lithuania, Latvia, Hungary, Poland, Czech Republic). Questionnaire interviews were used to collect data on self-rated health in the last 12 months, education, marital status, perceived control based on nine questions, and material deprivation based on availability of food, clothing and heating. For each population, two ecological measures of material inequalities were available: an inequality score estimated from the survey data as the distance between the 90th and 10th percentiles of material deprivation, and Gini coefficient from published sources. Data on 5330 men and women aged 20-60 were analysed. Prevalence of poor health (worse than average) varied between 8% in Czechs and 19% in Hungarians. The age-sex-adjusted odds ratio for university vs primary education was 0.36 (0.26-0.49); odds ratios per 1 standard deviation increase in perceived control and in material deprivation were 0.58 (95% CI 0.48 0.69) and 1.51 (1.40-1.63), respectively. The odds ratio for an increase in inequality equivalent to the difference between the most and the least unequal populations was 1.49 (0.88-2.52) using the material inequality score and 1.41 (0.91-2.20) using the Gini coefficient. No indication of an effect of either inequality measure was seen after adjustment for individuals' deprivation or perceived control. The results suggest that, as in western populations, education and material deprivation are strongly related to self-rated health. Perceived control appeared statistically to mediate some of the effects of material deprivation. The non-significant effects of both ecological measures of inequality were eliminated by controlling for individuals' characteristics. PMID- 11037222 TI - Psychosocial risk factors, inequality and self-rated morbidity in a changing society. AB - The aim of this study was to analyse the interaction of social, economic, psychological and self-rated health characteristics of the Hungarian population in representative, stratified nation-wide samples during the period of sudden political-economic changes. In 1988 20,902 and in 1995 12,640 persons, representing the Hungarian population over the age of 16 by age, sex and place of residence were interviewed. Self-rated morbidity characteristics, shortened Beck Depression Inventory, hopelessness, hostility, ways of coping, social support, control over working situation and socioeconomic characteristics were examined. Age dependent changes could be observed between 1988 and 1995 with increasing depressive symptomatology, hopelessness, lack of control over working situation in the population above 40 years, while in the younger population improvements in depressive symptomatology could be seen. According to hierarchical loglinear analysis, depressive symptom severity mediates between relative socioeconomic deprivation and higher self-rated morbidity rates, especially among men. Depressive symptomatology is closely connected with hostility, low control in working situation, low perceived social support and emotional ways of coping. A vicious circle might be hypothesised between socially deprived situation and depressive symptomatology, which together has a major role in higher self-rated morbidity rates. PMID- 11037223 TI - Educational differences in self-rated health during the Russian transition. Evidence from Taganrog 1993-1994. AB - In the beginning of 1992, in order to do away with the ruins of the old communist system once and for all, radical economic reforms--'shock therapy'--were introduced in Russia. However, there are winners and losers in the Russian transition from communism to market economy and democracy. The aim of the study is to investigate whether there are educational differences in self-rated health among the citizens of Taganrog in 1993-94. If there are indeed differences, the second aim is to investigate whether they can be explained by variations in specific social, economic and psychological circumstances, which are known to be affected by the present social and economic transformation in Russia. The analysed survey was carried out in a middle-sized Russian city, Taganrog, in late 1993 and early 1994. It was conducted by means of face-to-face interviews, with a sampling frame consisting of dwellings selected from an official register and stratified by type and size. The analysed sample consists of 2372 respondents, aged 25-54 years, in 1414 households. Data were analysed in logistic regressions with self-rated health as a dichotomised outcome variable. The results show that those with less than compulsory education reported poor health twice as often as the highest-educated group. Material prosperity and relations within the family were important for self-rated health and explained to a certain degree the educational health differences. Lower educational groups now live under conditions often characterised by economic hardship, family conflicts, etc., and consequently also poor health. PMID- 11037224 TI - Self-rated health and occupational conditions in Russia. AB - A low level of health promotion and disease prevention awareness in Russia, such as disregard for personal health during periods of well-being or illness, have contributed to a decline in general population health over the past decade. The relationship of working conditions and health awareness was explored among a sample of adults in Russia. This research project was conducted, July-August, 1998, in the city of Kazan. Data was collected by probability sampling of addresses and personal interview. Working conditions influence general public and family health. The occupation of the spouse contributed significantly to family conflicts, and financial problems ranked first as the cause of marital conflict. Due to lack of material resources, one-third of respondents, even though employed, reported being dissatisfied with the quality of their nutritional status and one-fifth with the lack of leisure time. Although more than two-thirds were satisfied with their work, every fifth did not consider their wages sufficient, every fourth wanted to change occupations, and every third was afraid of being fired. The majority of employed respondents reported low salaries, worked a full 8-hour day at their primary occupations and one-fifth had a second job. Younger people were especially prone to disregard their health with intensive work schedules. About half of respondents reported being exposed to toxic health hazards in their past or current jobs. Almost one-fifth agreed to work at hazardous occupations because of higher salaries. Social status, indicated by a higher education, was associated with having the opportunity to chose work in more favourable circumstances, and consequently with less health risks. In summary, the research demonstrated that health was an instrumental value in Russia, exploited as a economic resource not only during periods of well being but also during illness, by individuals not seeking preventive or timely health care because of the fear of losing their jobs. PMID- 11037225 TI - The HIV/AIDS epidemic in Ukraine: its potential social and economic impact. AB - Present and immediately foreseeable medical knowledge suggest that HIV infection cannot be avoided by vaccination and that an affordable cure for the resulting syndrome, AIDS, is a long way off. There is a strong possibility that Ukraine is confronted by an HIV epidemic which will spread into the general population and that the most common mode of transmission will be through heterosexual intercourse. The epidemic in the Ukraine is currently concentrated among intravenous drug users. It is estimated that between 60,000 and 180,000 people may currently be infected. In present economic and social circumstances there are many features of Ukrainian society that may add to the probability of the epidemic becoming widespread in the general population. It is likely that this process may have already commenced. The result of this will be numerous additional deaths and illness over the short (5 year) (19,000-23,000 deaths), medium (10-15 year) (61,000-111,000), and longer terms (>20 year) (in excess of 40,000-160,000 deaths). The research reported here was undertaken in 1997-8 and describes the potential medium to long term social and economic impact of an HIV/AIDS epidemic in Ukraine. Using the concepts of risk environment, susceptibility and vulnerability, it reports the problems which might be expected to develop in relation to care of excess orphans, the elderly, vulnerable households and regions as well as among those working in the "third sector", a social sector upon which exponents of the importance of developing sound "civil society" in "transitional economies" place heavy emphasis. PMID- 11037226 TI - Eastern European transition and suicide mortality. AB - The current paper seeks to systematize the discussion on the causes of the changes in Eastern European countries' suicide mortality during the last 15 years by analyzing the changes in relation to some common causes: alcohol consumption, economic changes, "general pathogenic social stress", political changes, and social disorganization. It is found that the developments in suicide have been very different in different countries, and that the same causes cannot apply to all of them. However, the relation between suicide mortality and social processes is obvious. A model consisting of the hypothetical general stress (as indicated by mortality/life expectancy), democratization, alcohol consumption, and social disorganization (with a period-dependent effect) predicted the percentual changes in the suicide rates in 16 out of the 28 Eastern Bloc countries in 1984-89 and 1989-94 fairly accurately, while it failed to do this for Albania, Poland, Romania, Slovakia, and the Caucasian and Central Asian newly independent states. Most interesting were the strong roles played by changes in life expectancy, the causes of which are discussed, and the fact that economic change seemed to lack explanatory power in multiple analyses. The data are subject to many potential sources of error, the small number of units and the large multicollinearity between the independent variables may distort the results. Nevertheless, the results indicate that the changes in Eastern European suicide mortality, both decreases and increases, may be explained with the same set of variables. However, more than one factor is needed, and the multicollinearity will continue to pose problems. PMID- 11037227 TI - How much does social capital add to individual health? A survey study of Russians. AB - Some Russians are healthier than others. To what extent does their health vary with involvement or exclusion from social capital networks? The first section reviews alternative theories: human capital as the primary determinant; social capital, whether generic, situation-specific or simply a new label for old measures of social integration; or a composite theory--both human and social capital are major determinants of health. The evidence to test hypotheses consists of individual-level data about self-assessed physical and emotional health from the special-purpose social capital questionnaire used in the 1998 New Russia Barometer survey, a nationwide representative sample of the adult Russian population. Multiple regression analysis shows that on their own human capital and social capital each account for a notable amount of variance in health. When both forms of capital are combined in a composite model, each retains major influence, demonstrating that social capital does make an independent contribution to health. Significant social capital influences include involvement or exclusion from formal and informal networks; friends to rely on when ill; control over one's own life; and trust. Significant human capital influences besides age include subjective social status, gender and income. Regression-based estimates of impact show that social capital increases physical and emotional health more than human capital; together they can easily raise an individual's self-reported health from just below average on a five-point scale to approaching good health. PMID- 11037228 TI - Changes in cow teat tissue created by two types of milking cluster. AB - We have investigated the responses of cow teats to machine milking in a study of relatively newly installed commercial milking parlours fitted with one of two types of milking cluster. The first was a common type with a large claw volume (> 200 ml), 15-16 mm i.d. long milk tube, 10 mm short pulse tube, cluster weight < 3 2 kg and used alternate pulsation. The second was a more traditional type with a 150 ml claw bowl volume, 13.5 mm i.d. long milk tube, 8 mm short pulse tube, cluster weight approximately 3.5 kg and used simultaneous pulsation. We scored approximately 50 cows in each of 20 herds, all within 60 s of cluster removal, for changes from the premilking teat condition: teat colour (creation of reddening or blueness), firmness, thickening at the base of the teat associated with the position of the liner mouthpiece, and whether the teat duct orifice was open. There were statistically significant differences in the proportion of cows displaying these four alterations in teat condition between herds using the two types of cluster. The more common type of cluster was always associated with better teat condition. The cause and effect of poorer teat condition have not been fully established and are likely to be multifactorial. The principal risk factors may be cluster weight, overmilking, vacuum applied during the overmilking phase and the design of the liner mouthpiece. PMID- 11037229 TI - Effects of teatcup liner tension on teat canal keratin and teat condition in cows. AB - The effect of tension of teatcup liners on teat end condition and quantity of keratin in the teat canal was investigated. Liner tension was increased by using longer teatcup shells. The first experiment used six Holstein cows in early lactation. Left quarters were milked with liners under medium or normal tension by using Conewango liners in 142 mm shells. Right quarters were milked with liners under high tension by mounting the liners in teatcup shells 149 mm in length. By day 16 teat end condition and sensitivity to manipulation were worsened by thrice daily milking when liners were under a higher tension. Two subsequent experiments each used 12 different Holstein cows. These cows were in mid lactation and were milked twice daily for 10 or 30 d. Left quarters were milked with liners under high tension Right quarters were milked with liners under low tension by using teatcup shells 126 mm in length. The quantity of keratin removed during milking was not influenced by liner tension: however, the quantity of keratin at the end of the experiments was increased 10-20% in teats that were milked using liners under higher tension. Histological analysis and keratin content were consistent with epithelial hyperplasia induced by milking with liners under increased tension. PMID- 11037230 TI - Comparison of heat and pressure treatments of skim milk, fortified with whey protein concentrate, for set yogurt preparation: effects on milk proteins and gel structure. AB - Heat (85 degrees C for 20 min) and pressure (600 MPa for 15 min) treatments were applied to skim milk fortified by addition of whey protein concentrate. Both treatments caused > 90 % denaturation of beta-lactoglobulin. During heat treatment this denaturation took place in the presence of intact casein micelles; during pressure treatment it occurred while the micelles were in a highly dissociated state. As a result micelle structure and the distribution of beta lactoglobulin were different in the two milks. Electron microscopy and immunolabelling techniques were used to examine the milks after processing and during their transition to yogurt gels. The disruption of micelles by high pressure caused a significant change in the appearance of the milk which was quantified by measurement of the colour values L*, a* and b*. Heat treatment also affected these characteristics. Casein micelles are dynamic structures, influenced by changes to their environment. This was clearly demonstrated by the transition from the clusters of small irregularly shaped micelle fragments present in cold pressure-treated milk to round, separate and compact micelles formed on warming the milk to 43 degrees C. The effect of this transition was observed as significant changes in the colour indicators. During yogurt gel formation, further changes in micelle structure, occurring in both pressure and heat-treated samples, resulted in a convergence of colour values. However, the microstructure of the gels and their rheological properties were very different. Pressure-treated milk yogurt had a much higher storage modulus but yielded more readily to large deformation than the heated milk yogurt. These changes in micelle structure during processing and yogurt preparation are discussed in terms of a recently published micelle model. PMID- 11037231 TI - Chromatographic characterization of ovine kappa-casein macropeptide. AB - Ovine casein macropeptide (CMP) was characterized by anion-exchange FPLC and reversed-phase (RP) HPLC. To study heterogeneity (the degree of glycosylation and phosphorylation), CMP was desialylated with neuraminidase and dephosphorylated with acid phosphatase. Following RP-HPLC, the main CMP components were identified using either on-line or off-line mass spectrometry. The most abundant ovine CMP component was a diphosphorylated carbohydrate-free form, followed by one or two monophosphorylated and a non-phosphorylated asialo-aglyco species. Aglyco non phosphorylated, monophosphorylated and diphosphorylated forms were in the ratio 3:20:77. Only approximately 30% of ovine CMP was glycosylated. Assuming that the monosaccharide fraction of ovine CMP is composed of N-acetylgalactosamine, galactose and N-glycolylneuraminic acid, molecular masses consistent with the presence of CMP containing tetra-, tri-, di- and monosaccharide were identified. PMID- 11037232 TI - Mineral and casein equilibria in milk: effects of added salts and calcium chelating agents. AB - We have investigated the effects of adding a range of mineral salts and calcium chelating agents on the distribution of casein and minerals between the non pelleted and pelleted phases of milk obtained upon centrifugation at 78000 g for 90 min. Adding CaCl2 or mixtures of NaH2PO4 and Na2HPO4 to reconstituted skim milk (90 g milk solids/kg) at pH 6.65 increased both pelleted casein and pelleted calcium phosphate. Opposite effects were obtained by adding citrate or EDTA. The change in pelleted calcium phosphate was not simply related to casein release from the micelle. Upon adding 5 mmol EDTA/kg milk, 20% of the pelleted Ca, 22% of the pelleted phosphate and 5% of the micellar casein were removed. Increasing the concentration of EDTA to 10 mmol/kg milk decreased the pelleted Ca by 44% and the pelleted phosphate by 46%, and caused 30% of the micellar casein to be released. The effects of adding phosphate, citrate or EDTA at pH 6.65, followed by the addition of CaCl2, demonstrated the reversibility of the dissolution and formation of the micellar calcium phosphate. There were limits to this reversibility that were related to the amount of colloidal calcium phosphate removed from the casein micelles. Adding CaCl2 to milk containing > or = 20 mmol EDTA or > or = 30 mmol citrate/kg milk did not result in complete reformation of casein micelles. Light-scattering experiments confirmed that the dissolution of moderate amounts of colloidal calcium phosphate had little effect on micellar size and were reversible, while the dissolution of larger amounts of colloidal calcium phosphate resulted in large reductions in micellar size and was irreversible. PMID- 11037233 TI - Flavour sulphides are produced from methionine by two different pathways by Geotrichum candidum. AB - We have investigated the capacities of Geotrichum candidum strains to produce sulphides from methionine. This attribute is very important in cheese technology because of the flavouring potential of sulphur compounds. A spectrophotometric procedure using 5,5'-dithiobis(2-nitrobenzoic acid) to determine sulphides was tested on a collection of G. candidum strains, and confirmed by gas chromatography-mass spectrometry. The strains were distinguished on the basis of their ability to produce methanethiol. Gas chromatography-mass spectrometry also made it possible to identify other sulphides, such as dimethyl disulphide, dimethyl trisulphide and dimethyl sulphide. Using sonicated cells, the specific production of these four sulphides was studied in presence of L-[S-methyl 2H]methionine. Both methanethiol and dimethyl sulphide were produced from methionine, but two different pathways were used by G. candidum. PMID- 11037234 TI - DNA fingerprinting of thermophilic lactic acid bacteria using repetitive sequence based polymerase chain reaction. AB - DNA fingerprints of lactic acid bacteria were generated by polymerase chain reaction using a primer based on the repetitive elements found in the genome of Streptococcus pneumoniae (BOX-PCR). The method made it possible to identify 37 isolates from raw milk. industrial starters and yogurt. Differentiation at species, subspecies and strain level was possible for Lactobacillus delbrueckii subsp. lactis, Lb. delbrueckii subsp bulgaricus and Str. thermophilus. BOX-PCR was also applied to studying the strain composition of a starter culture and the direct detection of strains in commercial fermented milk. PMID- 11037235 TI - Coagulating and lipolytic activities of artisanal lamb rennet pastes. AB - Lamb rennet pastes were prepared by the procedure most commonly used by Idiazabal cheese manufacturers. We studied the effects on their coagulating and lipolytic activities of the state of the stomach at the time of death (full of milk or empty), the amount of NaCl added, the origin of the lambs and paste storage time. Coagulating activities were generally between 155 and 363 units/g tissue. Pastes prepared from stomachs of lambs from slaughterhouse flocks had significantly higher coagulating activities than those of lambs from separate flocks. No significant decrease in coagulating activity was observed after 1 year storage at 4 degrees C. Chymosin represented 75-80% of the total coagulating activity with the remainder being pepsin. Rennet paste extracts with pH < 4.7 did not have increased coagulating activities when their pH was lowered to 2.0, while those with pH > 5.2 had activities 1.5-fold those before treatment. Lipase activity was higher in extracts of rennet pastes prepared using the stomachs of lambs that arrived at the slaughterhouse in the morning just prior to slaughter than in those prepared with the stomachs of lambs that had arrived on the previous evening. However, the reverse was the case for esterase activity. Activating the coagulating activity by pH cycling completely destroyed both lipolytic activities. Storage at 4 degrees C for > 1 year did not affect esterase activity but lipase activity decreased substantially after 4-5 months. Lipase, but not esterase, activity was responsible for the liberation of short-chain free fatty acids from ovine milk fat. PMID- 11037236 TI - Standardized reaction times used to describe the mechanism of enzyme-induced gelation in whey protein systems. AB - Whey protein isolate (WPI), either untreated or pretreated at 80 degrees C for 30 min, was incubated with a proteinase from Bacillus licheniformis until a gel was formed. Standardized reaction times, directly linked to the degree of hydrolysis, were obtained from plots of the relative amount of peptides released v. reaction time obtained under different conditions (enzyme concentration, temperature, pH, NaCl addition). This provided a connection between the gelation profile and the degree of hydrolysis. In the case of untreated WPI, gelation occurred at lower degrees of proteolysis when the enzyme concentration was decreased, demonstrating that a rate-limiting aggregation process occurred at the same time as the proteolysis in a manner similar to the renneting of milk. This was not the case for preheated WPI, when gelation was found to take place at a constant degree of proteolysis, independent of the enzyme concentration. In this case, the mechanism could be described by assuming the thermally induced aggregates present in this substrate had progressively more stabilizing peptide segments shaved off, resulting in increased attraction between individual aggregates that ultimately led to gelation. Results obtained at 40-60 degrees C supported this, as we found no effect of temperature on the degree of proteolysis at gelation for the untreated WPI, whereas the degree of proteolysis decreased with increasing temperature when heated WPI was hydrolysed. The effect of pH and NaCl addition on the process was to reduce repulsion between the aggregating species so that gelation was induced at a decreased degree of proteolysis. PMID- 11037237 TI - Rheological properties of milk gels formed by a combination of rennet and glucono delta-lactone. AB - The effects of heat treatment of milk, and a range of rennet and glucono-delta lactone (GDL) concentrations on the rheological properties, at small and large deformation, of milk gels were investigated. Gels were made from reconstituted skim milk at 30 degrees C, with two levels each of rennet and GDL. Together with controls this gave a total of sixteen gelation conditions, eight for unheated and eight for heated milk. Acid gels made from unheated milks had low storage moduli (G') of < 20 Pa. Heating milks at 80 degrees C for 30 min resulted in a large increase in the G' value of acid gels. Rennet-induced gels made from unheated milk had G' values in the range approximately 80-190 Pa. However, heat treatment severely impaired rennet coagulation: no gel was formed at low rennet levels and only a very weak gel was formed at high levels. In gels made with a combination of rennet and GDL unusual rheological behaviour was observed. After gelation, G' initially increased rapidly but then remained steady or even decreased, and at long ageing times G' values increased moderately or remained low. The loss tangent (tan delta) of acid gels made from heated milk increased after gelation to attain a maximum at pH approximately 5.1 but no maximum was observed in gels made from unheated milk. Gels made by a combination of rennet and GDL also exhibited a maximum in tan delta, indicating increased relaxation behaviour of the protein-protein bonds. We suggest that this maximum in tan delta was caused by a loosening of the intermolecular forces in casein particles caused by solubilization of colloidal calcium phosphate. We also suggest that in combination gels made from unheated milk a low value for the fracture stress and a high tan delta during gelation indicated an increased susceptibility of the network to excessive large scale rearrangements. In contrast. combination gels made from heated milk formed firmer gels crosslinked by denatured whey proteins and underwent fewer large scale rearrangements. PMID- 11037238 TI - Relatedness of Staphylococcus aureus isolates from bovine mammary gland suffering from mastitis in a single herd. PMID- 11037239 TI - Composition of the sterol fraction of caprine milk fat by gas chromatography and mass spectrometry. PMID- 11037240 TI - Effect of beta-lactoglobulin polymorphism on milk-related traits of dairy ewes analysed by a repeated measures design. PMID- 11037241 TI - Action of cardosin A from Cynara humilis on ovine and caprine caseinates. PMID- 11037242 TI - Involvement of a pasteurizer in the contamination of milk by Bacillus cereus in a commercial dairy plant. PMID- 11037243 TI - Lipolysis and oxidative stability of soft ripened cheeses containing vegetable oils. PMID- 11037244 TI - MMR and the measles crisis. PMID- 11037245 TI - Salt and blood pressure. PMID- 11037246 TI - General practice. PMID- 11037247 TI - Health and homelessness in Dublin. AB - This study examined health and related issues in the adult homeless population of Dublin. The data was collected over a five day period by trained interviewers using an anonymous questionnaire. The response rate was 64%. The population was made up of different demographic sub-groups. Almost 80% were smokers, 30% drank alcohol beyond recommended limits and 30% used illegal drugs. Almost half perceived their health as poor. Sixty six per cent of people had at least one physical or psychiatric problem. Chronic disease was reported by 41%. Health problems varied with demographic and behavioural factors. Utilisation of services varied with age, sex and other demographic factors. Many barriers to service utilisation were identified. Homeless people have increased risks for illness and suffer similar, but more prevalent, health problems to the general population for which they do not always receive adequate or appropriate care. Recommendations include further research and specific service developments. PMID- 11037248 TI - Establishing a primary care based anticoagulation clinic. PMID- 11037249 TI - The acceptability of pneumococcal vaccine to older persons in Ireland. AB - A study was carried out to demonstrate in an Irish population whether older persons at risk from pneumococcal disease would accept an offer of the pneumococcal vaccine at the same time as their influenza vaccination. Of the 450 patients from 2 practices invited to attend for vaccination, 367 (81.6%) accepted both vaccines, a further 17 (3.8%) accepted the influenza vaccine only and a further 3 (0.7%) accepted the pneumococcal vaccine only. Three hundred and seven (68.2%) patients had received influenza vaccine at some time in previous years and these were statistically more likely to accept either vaccine than those who had not. This study has demonstrated that older persons at risk from pneumococcal disease will respond positively to a written invitation from their GP to avail of the pneumococcal vaccine. PMID- 11037250 TI - A cost-benefit evaluation of helicopter transfers to the Beaumont neurosurgical unit. AB - A retrospective analysis of all consecutive helicopter transfers to the Beaumont neurosurgical unit was performed over a two year period from January 1994 to December 1996. There were 55 transfers, 55% of which were for trauma patients. Sixty two per cent were intubated prior to transfer. The mean length of stay at the referring hospital prior to transfer was 12.25 hours (range 3-98 hours). Twenty five per cent underwent a neurosurgical operation within 2 hours of arrival at the unit. Seventy eight per cent of patients either made a full recovery or were left with mild to moderate disability. Whilst helicopters do not seem to result in a marked reduction in total transfer time compared with 'blue light' ambulances, they do result in a shorter time being spent in the unstable transfer environment and this, in turn, might be associated with improved patient outcome. PMID- 11037251 TI - Stress in an accident and emergency department. PMID- 11037252 TI - Lupus support and awareness in the Republic of Ireland. PMID- 11037253 TI - Effectiveness of mask ventilation performed by hospital doctors in an Irish tertiary referral teaching hospital. AB - The objective of this study was to assess the effectiveness of mask ventilation performed by 112 doctors with clinical responsibilities at a tertiary referral teaching hospital. Participant doctors were asked to perform mask ventilation for three minutes on a Resusci Anne mannequin using a facemask and a two litre self inflating bag. The tidal volumes generated were quantified using a Laerdal skillmeter computer as grades 0-5, corresponding to 0, 334, 434, 561, 673 and > 800 ml respectively. The effectiveness of mask ventilation (i.e. the proportion of ventilation attempts which achieved a volume delivery of > 434 mls) was greater for anaesthetists [78.0 (29.5)%] than for non anaesthetists [54.6 (40.0)%] (P = 0.012). Doctors who had attended one or more resuscitation courses where no more effective at mask ventilation than their colleagues who had not undertaken such courses. It is likely that first responders to in-hospital cardiac arrests are commonly unable to perform adequate mask ventilation. PMID- 11037254 TI - The other form of ADR operated in US health care: binding arbitration. PMID- 11037255 TI - Counting red cells--is it an answer to EIPH ? PMID- 11037256 TI - What can we learn by growing equine cells in culture? PMID- 11037257 TI - Superficial digital flexor tendonitis in the horse. AB - The superficial digital flexor tendon (SDFT) is an elastic structure that during maximal exercise appears to operate close to its functional limits. The biomechanical and biochemical responses to exercise, injury, and healing are still poorly understood but ongoing research is providing valuable new information which is addressed in this review. It appears that the SDFT matures early, after which time it has limited ability to adapt to stress and undergoes progressive degeneration. Focal hypocellularity, collagen fibril degeneration, selective fibril loading and alterations in the noncollagenous matrix occur primarily within the central core region of the midmetacarpal segment. Current treatment strategies have had equivocal results in returning animals to optimal athletic activity. To date it would seem that progressive rehabilitation programmes coupled with regular ultrasonographic evaluations are a cost-effective and comparable strategy when compared to surgical treatment methods. Recent interest in pharmacological modulation of intrinsic healing of collagenous structures has led to the investigation of various growth factors as potential therapeutic aids in the healing of tendon injuries. However, one of the major goals in tendon research, and one which holds the most optimism for success in the immediate future, is the prevention of tendon injuries. PMID- 11037258 TI - Relationship of pulmonary arterial pressure to pulmonary haemorrhage in exercising horses. AB - Exercise-induced pulmonary haemorrhage (EIPH) is characterised by blood in the airways after strenuous exercise and results from stress failure of the pulmonary capillaries. The purpose of this experiment was to establish a threshold value of transmural pulmonary arterial pressure at which haemorrhage occurs in the exercising horse. Five geldings, age 4-14 years, were run in random order once every 2 weeks at 1 of 4 speeds (9, 11, 13, 15 m/s); one day with no run was used as a control. Heart rate, pulmonary arterial pressure and oesophageal pressure were recorded for the duration of the run. Transmural pulmonary arterial pressure was estimated by electronic subtraction of the oesophageal pressure from the intravascular pulmonary arterial pressure. Within 1 h of the run, bronchoalveolar lavage was performed and the red and white blood cells in the fluid were quantified. Red cell counts in the lavage fluid from horses running at 9, 11 and 13 m/s were not significantly different from the control value, but after runs at 15 m/s, red cell counts were significantly (P<0.05) higher. White cell counts were not different from control values at any speed. Analysis of red cell count vs. transmural pulmonary arterial pressure indicated that haemorrhage occurs at approximately 95 mmHg. Red cell lysis in the lavage fluid was also apparent at transmural pulmonary arterial pressures above 90 mmHg. We conclude that, in the exercising horse, a pulmonary arterial pressure threshold exists above which haemorrhage occurs, and that pressure is often exceeded during high speed sprint exercise. PMID- 11037259 TI - Anaemia, diarrhoea and opportunistic infections in Fell ponies. AB - This report summarises clinical and pathological observations on Fell pony foals with a range of signs that included ill thrift, anaemia, respiratory infection, glossal hyperkeratosis and diarrhoea. Some of the foals had normochromic, normocytic anaemia and some had low levels of plasma proteins, including immunoglobulin G. Antibiotic and supportive treatment was ineffective and all affected foals died or were killed on humane grounds. Postmortem examination of 12 foals and tissues from 2 other foals revealed a range of lesions that included glossal hyperkeratosis, typhlocolitis, intestinal cryptosporidiosis, granulomatous enteritis, proliferative and necrotising bronchiolitis consistent with adenovirus infection; lesions similar to those in the respiratory tract were present in the salivary gland and pancreas of individual foals. Lymphoid tissue was judged to be smaller than expected. These observations suggest the possibility of opportunistic infections secondary to some form of undefined immunocompromised state. PMID- 11037260 TI - Relationship between clinical signs and lung function in horses with recurrent airway obstruction (heaves) during a bronchodilator trial. AB - During a trial to determine the dose response to the beta2-adrenergic agonist pirbuterol, we judged the severity of airway obstruction by use of a clinical scoring system and compared this to objective data obtained by quantitative measures of lung function. Six horses affected by recurrent airway obstruction were used in this trial. Four hundred and sixty-eight measurements of lung function and clinical scores were obtained from 13 measurement periods when horses received each of 6 doses of pirbuterol. Scores of 1-4 were assigned to degree of nasal flaring and abdominal effort and summed for a total score. The veterinarian scoring the signs did not know the dose of pirbuterol received by the horse and was unaware of the lung function data. Nasal, abdominal and total scores were significantly related to changes in lung function and changes in breathing pattern. There were significant differences between total scores greater than 5 in indices that reflected changes in breathing strategy (peak inspiratory and expiratory flow), peripheral airway obstruction (dynamic elastance), and effort of breathing (maximal change in pleural pressure). Below a total score of 5, there were fewer significant differences in lung function even though measurements of pulmonary resistance and dynamic elastance indicated considerable airway obstruction. Failure of clinical score to reflect this low grade airway obstruction suggests that airway disease is underdiagnosed and its detection would be helped by the availability of a convenient lung function test. PMID- 11037261 TI - In vitro model of equine muscle regeneration. AB - Equine satellite cells are responsible for muscle healing and regeneration in the mature horse. We describe the in vitro cell culture conditions required for clonal populations of equine satellite cells to undergo both proliferation and differentiation. Our hypothesis is that these in vitro conditions model regeneration of muscle and can be used to evaluate potential therapeutics. In this study, 2 areas of satellite cell response were tested: proliferation of clones induced by growth factors, and fusion induced by culture conditions. Equine satellite cell clones showed differences in their response to growth factors as well as accumulation of cellular protein concentrations. Equine satellite cells proliferate in response to both human and bovine FGF. IGF-1, a powerful mitogen of other satellite cell culture systems, was not as effective for inducing equine satellite cell proliferation. Protein concentrations were also measured in satellite cell cultures. Clones differed in cellular protein produced depending on growth conditions. Conditions inducing differentiation into myotubes was also determined for a 96 well assay and can be used to study the final stage of functioning muscle production. This in vitro model is the first step in identifying potential therapeutics to speed wound healing and promote muscle regeneration in horses. PMID- 11037262 TI - Surgical treatment of 45 horses affected by squamous cell carcinoma of the penis and prepuce. AB - The medical records of 45 horses treated for suspected squamous cell carcinoma of the penis and/or prepuce were reviewed. The age of 40 horses was known, and these had a mean age of 17.4 years. The duration of neoplasia was known for only 3 of the 45 horses. The results of histological evaluation of lesions, available for 35 horses, confirmed that the diseased tissue was squamous cell carcinoma. The location of gross neoplastic lesions was recorded for 43 horses; the glans penis was involved in 24 horses, the body of the penis or the inner lamina of the preputial fold in 27 horses, and the external fold of the prepuce in 10 horses. Surgical treatments of these horses included phallectomy (penile amputation) in 35 horses, segmental posthectomy in 4 horses, phallectomy plus segmental posthectomy in 2 horses, and en bloc resection of the penis, prepuce and superficial inguinal lymph nodes with penile retroversion in 4 cases. Short-term complications in the immediate postoperative period included preputial oedema and haemorrhage at the end of urination. One horse developed acute urinary retention because of severe urethral oedema. Long-term (>1 year) follow-up information was obtained for 31 horses. Neoplasia of the penis and/or prepuce recurred in 6 of these 31 horses (19%), and in 5 of these the recurrence necessitated euthanasia of the horse. PMID- 11037263 TI - Tracheobronchial mucus viscoelasticity during environmental challenge in horses with recurrent airway obstruction. AB - The goal of this study was to compare the rheological properties of mucus from horses with recurrent airway obstruction (RAO) to that from healthy controls during environmental challenge by stabling in stalls with straw as bedding and hay as feed. We determined viscoelasticity (log G* dyn/cm2, at 10 radian/s) and calculated mucociliary clearability index (MCI) and cough clearability index (CCI), which are derivative parameters of G* and the ratio of viscosity and elasticity measured at 1 and 100 radian/s, respectively. We also investigated the solids content of mucus, and cytology of bronchoalveolar lavage fluid (BALF). Samples were obtained before (0 h) and 6, 24 and 48 h after environmental challenge. The central findings were rheological changes in airway mucus, which occurred over time in RAO-affected animals, but not in controls. Mucus rheology was similar in both groups at 0 and 6 h. In RAO-affected horses, mucus viscoelasticity, as measured by log G*, increased from 2.49 +/- 0.18 dyn/cm2 (mean +/- s.e.) at 0 h to 3.05 +/- 0.13 dyn/cm2 at 24 h after environmental challenge, and was accompanied by significant decreases in MCI and CCI. Percent solids of mucus did not differ significantly between the 2 groups, nor over time. Rheological values did not correlate with BALF cytology. We conclude that viscoelastic properties of tracheal mucus samples from RAO horses in remission do not differ from those of normal horses. However, environmental challenge causes clinical signs of small airway disease and a concurrent increase in mucus viscoelasticity only in RAO horses. Therefore, we infer that unfavourable changes in mucus rheology may contribute to stasis and accumulation of mucus in RAO horses in exacerbation, but not in clinical remission. PMID- 11037265 TI - Haemodynamic response to exercise in Standardbred trotters with red cell hypervolaemia. AB - In order to evaluate the haemodynamic response to exercise in Standardbred trotters with red cell hypervolaemia (RCHV), 12 trotters with RCHV were compared with 9 normovolaemic (NV) trotters. Haemodynamic data were recorded during exercise at 4 different speeds on a treadmill. Oxygen uptake was determined with an open bias flow system. Pulmonary artery pressure (PAP), systemic artery pressure (SAP), heart rate, packed cell volume (PCV) and plasma lactate and haemoglobin ([Hb]) concentrations were measured. Arteriovenous O2 content difference, cardiac output, stroke volume, pulmonary vascular resistance (PVR) and total systemic resistance (TSR) were calculated. Oxygen uptake, arteriovenous O2 content difference, heart rate, cardiac output, stroke volume, TSR and lactate did not differ between groups. The RCHV horses had significantly higher both mean diastolic and systolic PAP compared to NV horses and this difference increased with higher workload. Further, a higher SAP, PVR, PCV and [Hb] were found in RCHV horses during the course of exercise. Eleven of the RCHV horses, but none of the NV, showed exercise-induced pulmonary haemorrhage on endoscopic examination. The increase in red cell volume, resulting in a high PCV and high total blood volume, is suggested to be an important contributor to both the increased blood pressures in pulmonary and systemic circulation during exercise and to the development of exercise-induced pulmonary haemorrhage in RCHV horses. PMID- 11037264 TI - Equine proliferative enteropathy: a cause of weight loss, colic, diarrhoea and hypoproteinaemia in foals on three breeding farms in Canada. AB - Proliferative enteropathy (PE) is a transmissible enteric disease caused by Lawsonia intracellularis. An outbreak of equine PE was diagnosed in foals from 3 breeding farms. Most foals had been weaned prior to the appearance of clinical signs, which included depression, rapid and marked weight loss, subcutaneous oedema, diarrhoea and colic. Poor body condition with a rough haircoat and a potbellied appearance were common findings in affected foals. Respiratory tract infection, dermatitis and intestinal parasitism were also found in some foals. Haematological and plasma biochemical abnormalities included hypoproteinaemia, transient leucocytosis, anaemia and increased serum creatinine kinase concentration. Postmortem diagnosis of PE was confirmed on 4 foals based on the presence of characteristic intracellular bacteria within the apical cytoplasm of proliferating crypt epithelial cells of the intestinal mucosa, using silver stains, and by results of PCR analysis and immunohistochemistry. Antemortem diagnosis of equine PE was based on the clinical signs, hypoproteinaemia and the exclusion of common enteric infections. Faecal PCR analysis was positive for the presence of L. intracellularis in 6 of 18 foals tested while the serum of all 7 foals with PE serologically evaluated had antibodies against L. intracellularis. Most foals were treated with erythromycin estolate alone or combined with rifampin for a minimum of 21 days. Additional symptomatic treatments were administered when indicated. All but one foal treated with erythromycin survived the infection. This study indicates that equine PE should be included in the differential diagnosis of outbreaks of rapid weight loss, diarrhoea, colic and hypoproteinaemia in weanling foals. PMID- 11037266 TI - Environment and prednisone interactions in the treatment of recurrent airway obstruction (heaves). AB - Recurrent airway obstruction (RAO) or heaves is a manifestation of a hypersensitivity to dust, moulds, and spores in the environment of a susceptible horse. Although in the majority of RAO-affected horses, clinical remission can be achieved by keeping horses at pasture to reduce their allergen exposure, this often is not practicable. For this reason, we investigated if changing the environment of a single stall in a 4 stall stable was sufficient to improve lung function and reduce inflammation in RAO-affected horses. In addition, we determined if addition of oral prednisone provided additional benefit. Twelve RAO susceptible horses were stabled, fed hay, and bedded on straw until they developed airway obstruction. At this point, bedding was changed to wood shavings and they were fed a pelleted diet for 2 weeks. Lung function was measured and bronchoalveolar lavage was performed before and 3, 7, and 14 days after environmental modification. In a crossover design, horses were treated for the 14 days with prednisone tablets (2.2 mg/kg bwt, q. 24 h). Horses then returned to pasture for 30 days. Airway obstruction was greatest before environmental modification. Significant improvement in lung function occurred within 3 days of the change in environment and continued to Day 7. Airway function was best after 30 days at pasture. The clinical response achieved by environmental modification was not significantly improved by addition of oral prednisone. The total number of cells, total neutrophils, and percent neutrophils was greatest before environmental modification. In the absence of prednisone, total and percent neutrophils did not decrease until Day 14 and total cell number until 30 days at pasture. In the presence of prednisone, total cells and total and percent neutrophils decreased by Day 3 and again at pasture. The fact that lung function can be improved within 3 days by environmental management alone emphasises the need for allergen reduction as the cornerstone of treatment of RAO. Although prednisone induced a more rapid reduction in airway inflammation, this was not associated with a more rapid improvement in airway function. PMID- 11037267 TI - Evidence supporting an increased presence of reactive oxygen species in the diseased equine joint. AB - Reactive oxygen species (ROS) are capable of degrading many components of the joint in the presence of insufficient antioxidant defences, and as a result have been implicated in the pathogenesis of joint disease in horses. However, to our knowledge, evidence of ROS occurring in diseased joints of horses has not been reported. The objective of this experiment was to compare differences in synovial fluid protein carbonyl content (as a marker of oxidative modification of synovial fluid proteins by ROS) and the antioxidant status of synovial fluid between clinically normal and diseased equine joints. Synovial fluid was collected from the metacarpophalangeal, metatarsophalangeal, carpal and tarsal joints of 4 horses, age 2-5 years, as controls, and from diseased joints (metacarpophalangeal, metatarsophalangeal, carpal, tarsal and/or femoropatellar) of 61 horses, age 2-5 years. Synovial fluid protein carbonyl content was higher (P<0.01) in diseased joints as compared to controls. Antioxidant status of synovial fluid from diseased joints was higher, but not significantly, than that of controls (P = 0.0595). These findings require further study to determine their contribution to the overall disease process. PMID- 11037268 TI - Cytological identification and quantification of testicular cell types using fine needle aspiration in horses. AB - Fifteen stallions of different breeds, age 3-11 years, had their right testicles evaluated by fine needle aspiration cytology (FNAC). Cytological analysis showed the following spermatogenic cell types: spermatogonia (1.6% +/- 1.1); spermatocyte I (3.4% +/- 2.2); spermatocyte II (0.8% +/- 0.7); early spermatids (25.5% +/- 9.5); late spermatids (37.0% +/- 9.3). Spermatozoal numbers were expressed as the spermatic index (SI = 31.5% +/- 8.5) and Sertoli cells were expressed as the Sertoli cell index (SEI = 20.9% +/- 17.0) (means +/- s.d). Identification of cell types was relatively easy and no immediate adverse effects of aspiration were noted. The results suggest that FNAC of testis may assist clinical diagnosis in the study of male equine infertility. PMID- 11037269 TI - Preoperative bending and twisting of a dynamic compression plate for the repair of tibial tuberosity fracture in the horse. PMID- 11037270 TI - Serum alpha-1-proteinase inhibitor concentration in 2 Quarter Horse foals with idiopathic pyogranulomatous panniculitis. PMID- 11037271 TI - Two cases of equine mucormycosis caused by Absidia corymbifera. PMID- 11037272 TI - Intervention and human rights. PMID- 11037273 TI - The impact on civilians of the bombing of Kosovo and Serbia. AB - Before the 1999 bombing, Kosovo was among the poorest regions in Europe, with low scores on indices of health care. After the war, housing for much of the population is below even basically acceptable standards and health care is disrupted, with serious risk of epidemic diseases. Societal disintegration has led to high levels of stress. In Serbia, also a poor country, which already suffered from high levels of pollution before the war, large amounts of several highly toxic chemicals were liberated into the environment by the bombing. Targeting of electrical generating plant, water treatment facilities, Danube bridges, railways and roads has adverse effects on civilian life, endangers health and seriously affects health care. The military strategy of attack on civilian infrastructures is a war on public health. PMID- 11037274 TI - Traumatic experiences and psychological reactions among women in Bosnia during the war. AB - Internally displaced women (N = 77) in Bosnia were interviewed before and after participating in group psychotherapy during the war in 1994 and 1995 to gain background information and assess stress exposure and stress reactions. Nearly all the women had experienced loss of family members, many acts of violence and mental health impairment. The participants had high scores on a symptom scale, in terms of intrusive, avoidance and arousal symptoms and somewhat lower on depression/powerlessness items. After participating in short-term group therapy they reported significant reduction in symptoms. The women who had experienced most traumatic events and had most symptoms, reported greatest reduction in symptomatology. Some of the implications of the findings are discussed and it is concluded that group therapy may be helpful in war conditions, even though traditional preconditions for psychotherapy are not present. PMID- 11037275 TI - Human radiation experiments: what price informed consent? AB - In 1992 a system of public access was established to more than three million pages of files in government depositories related to radiation experiments on US citizens, including children, pregnant women, and convicts, studying the effects of radioactive isotopes and testicular irradiation. The background to some of these studies is described; many were considered by Ethics Committees and the results published in the open literature after peer review. PMID- 11037276 TI - Disease and security: the effect of emerging and re-emerging diseases. AB - This article examines the role of emerging new and re-emerging diseases in the failure of the 'Health for All by 2000' concept. The gap between infection and control has widened instead of closing, which has definite military and security implications. It is argued that there is a direct link between disease and the social order, and that the traditional view of development first, followed by an increase in health status, must change. However, the political will and the necessary resources to combat emerging and re-emerging diseases are still lacking in many countries. Through the improper use of treatments and dosages, most notably antibiotics, new strains of previously eradicated diseases are appearing. Added to this are new patterns of relationships between man, his environment and the occurrence of disease leading to new diseases making their appearance. There is a direct relationship between war and disease, making disease something with political, military and security relevance. The aspect of intelligence which has the task of predicting the future, now has to take note of the occurrence of disease and its effects. PMID- 11037277 TI - The new millennium--do we have the courage of optimism? PMID- 11037278 TI - Learning from Kosovo: the future of humanitarian intervention. PMID- 11037279 TI - 2nd World Water Forum. AB - The world is facing a water crisis. About one billion people lack access to safe water and three billion to appropriate sanitation. There will probably be two billion extra people on the planet by 2025; so while today's shortages are for fresh water and sanitation, tomorrow's are predicted to be for growing crops. Already, 10% of world food production is drawing down underground water faster than the recharge rates. Water also threatens to cause conflicts between regions- a state of affairs that is expected to worsen as this century progresses. To raise public awareness of these issues and pressurize politicians to act, the World Water Commission (WWC) held a Forum in March 2000 in The Hague. The Forum collected together a huge range of people from governments, non-government organizations (NGOs), workers for the environment and development, private sector water providers, trades unions and scientific institutes. They all discussed a document (World Water Vision: A Water Secure World, referred to as the Vision) drawn up by the WWC as a basis for action. The Forum culminated in a meeting of ministers from over 140 governments. PMID- 11037280 TI - Bilateral receptive field neurons in the hindlimb region of the postcentral somatosensory cortex in awake macaque monkeys. AB - Single-neuron activities were recorded in the hindlimb region of the primary somatosensory cortex and part of area 5 in awake Japanese monkeys. A total of 1050 units were isolated from five hemispheres of four animals. Receptive fields (RFs) and submodalities were identified for 90% of isolated neurons in areas 3a and 3b. The percentage decreased as the recoding site moved to the more caudal areas. Deep or skin submodality neurons were dominant in area 3a or area 3b, respectively. Deep submodality neurons increased in more caudal areas and were the majority in areas 2 and 5. These observations were consistent with those in the hand and/or digit or arm and/or trunk region. The identified neurons were classified by their RF positions into four types: the foot, leg, foot and leg, or hindlimb and other body parts type. Among 831 identified neurons, 33 neurons had bilateral RFs, 14 had ipsilateral RFs, and the rest (N=784) had contralateral RFs. The relative incidence of neurons with bilateral or ipsilateral RFs among identified neurons was less than 1% in areas 3a, 3b, and 1, and 16% or 25% in areas 2 or 5, respectively. Within areas 2 and 5, the percentage of neurons with bilateral or ipsilateral RFs was significantly smaller in the foot type (5%) than in other RF types (24-57%). RFs of the foot type were on the sole or single toe but never on multiple toes. These observations contrasted with the previous findings that neurons with bilateral RFs were more frequently seen in the hand and/or digit region and that RFs on multiple digit tips were dominant there. The present study thus demonstrated that neurons with bilateral RFs do exist in the hindlimb region. Similarly to the forelimb region, they were found mostly in areas 2 and 5, the caudalmost areas of the postcentral gyrus and hierarchically higher stages in information processing. The relative paucity of neurons with bilateral RFs on the foot, especially those with RFs on multiple toes, may reflect functional differences between the foot and the hand. PMID- 11037281 TI - Crossed reciprocal inhibition evoked by electrical stimulation of the lamprey spinal cord. AB - Activation of a motoneuron pool is often accompanied by inhibition of the antagonistic pool through a system of reciprocal inhibition between the two parts of the neuronal network controlling the antagonistic pools. In the present study, we describe the activity of such a system in the isolated spinal cord of the lamprey, when a tonic motor output is evoked by extracellular stimulation (0.5-1 s train of pulses, 20 Hz) of either end of the spinal cord. With two electrodes symmetrically positioned in relation to the midline, stimulation with either of them separately elicited prolonged (1-5 s) ipsilateral ventral root activity. Activity could be abolished by stronger, simultaneously applied, stimulation of the contralateral side of the cord, suggesting that reciprocal inhibition between hemisegments operates when a tonic motor output is generated. Simultaneous stimulation of both sides of the spinal cord with a single electrode with a large tip (300-400 microm in diameter), positioned over the anatomical midline, elicited inconsistent right-side, leftside, or bilateral ventral root responses. A minor displacement (10-20 microm) to the left or right from the midline resulted in activation of ipsilateral motoneurons, whereas the contralateral motoneurons were silent. These findings indicate that a small asymmetry in the excitatory drive to the left and right spinal hemisegments can be further amplified by reciprocal inhibition between the hemisegments. Longitudinal splitting of the spinal cord along the midline resulted in reduced reciprocal inhibition between the hemisegments separated by the lesion. The reduction was proportional to the extent of the split. The inhibition was abolished when the split reached nine segments in length. From these experiments, the longitudinal distribution of the commissural axons responsible for inhibition of contralateral motor output could be estimated. PMID- 11037282 TI - Postural invariance in three-dimensional reaching and grasping movements. AB - The question of whether the final arm posture to be reached is determined in advance during prehension movements remains widely debated. To address this issue, we designed a psychophysical experiment in which human subjects were instructed to reach and grasp, with their right arm, a small sphere presented at various locations. In some trials the sphere remained stationary, while in others (the perturbed trials) it suddenly jumped, at movement onset, to a new unpredictable position. Our data indicate that the final configuration of the upper limb is highly predictable for a given location of the sphere. For movements directed at stationary objects, the variability of the final arm posture was very small in relation to the variability allowed by joint redundancy. For movements directed at "jumping" objects, the initial motor response was quickly amended, allowing an accurate grasp. The final arm posture reached at the end of the perturbed trials was neither different from nor more variable than the final arm posture reached at the end of the corresponding stationary trials (i.e. the trials sharing the same final object location). This latter result is not trivial, considering both joint redundancy and the motor reorganization imposed by the change in sphere location. In contrast to earlier observations, our data cannot be accounted for by biomechanical or functional factors. Indeed, the spherical object used in the present study did not constrain the final arm configuration or the hand trajectory. When considered together, our data support the idea that the final posture to be reached is planned in advance and used as a control variable by the central nervous system. PMID- 11037283 TI - Spatial and temporal aspects of eye-hand coordination across different tasks. AB - The way in which saccadic eye movements are elicited influences their latency and accuracy. Accordingly, different tasks elicit different types of saccades. Using the tasks steps, gap, memory, scanning and antisaccade, we analyzed combined eye and hand movements to determine whether both motor systems share control strategies. Errors and latencies were measured to examine whether changes in eye motor behavior are reflected in hand motor behavior. Directional and variable errors of eye and hand changed differently according to the tasks. Moreover, errors of the two systems did not correlate for any of the tasks investigated. Contrary to errors, mean latencies of eye movements were organized in the same pattern as hand movements. A correlation of latencies indicates that both motor systems rely on common information to initiate movement. Temporal coupling was stronger for intentional tasks than for reflexive tasks. PMID- 11037284 TI - Coordination disorders in patients with Parkinson's disease: a study of paced rhythmic forearm movements. AB - Whereas the consequences of Parkinson's disease (PD) for the performance of single-limb movements are well documented (i.e., bradykinesia, akinesia, rigidity, and tremor), fairly little is known about its implications for the coordination between limb movements. To help resolve this situation an experiment was conducted in which 11 PD patients and 11 control subjects performed rhythmic forearm movements at a comfortable amplitude in the in-phase, antiphase, and single-arm mode at pacing frequencies ranging from 0.5 to 3 Hz. The PD group displayed marked coordination problems over and above the known clinical motor symptoms of PD. The performance of both the in-phase and antiphase modes was significantly affected in the PD group compared to the control group; furthermore, the variability of relative phase was significantly increased in this group. These observations were not caused by problems to synchronize the movements with the external pacing signal. In addition to the bimanual coordination problems, involuntary mirror movements (MM) were observed in the single-arm control trials that were significantly larger in the PD group (4.4% of the amplitude of the moving arm) than in the control group (2.3%), suggesting a reduced ability to suppress a basic in-phase coupling of the arms. In the PD group, MM were largest during movements of the least-affected arm. These parkinsonian coordination problems are interpreted in terms of recent evidence on the neural organization of bimanual coordination, suggesting that they are due to cortical rather than callosal dysfunction. PMID- 11037285 TI - Postnatal development of differential projections from the caudal and rostral motor cortex subregions. AB - The primary motor cortex of cats, monkeys, and humans has distinct rostral and caudal subregions. In the cat, projections from the caudal subregion terminate predominantly in laminae 4-6 and, from the rostral subregion, in laminae 6-8. The purpose of this study was to determine if these distinctive termination patterns are present during early postnatal development, when corticospinal axons are establishing connections with spinal neurons, or if there was postnatal refinement of the distribution of terminations. We used the anterograde tracer biotinylated dextran amine (BDA) to label selectively projections from the two subregions in immature (postnatal days 25, 35 at time of analysis) and mature animals. We compared the distribution of spinal gray matter labeling from caudal and rostral motor cortex. In immature animals, substantial bilateral terminations were present after tracer injection into either subregion. Partial axon reconstructions revealed that individual axons terminated bilaterally. The dorso ventral laminar distribution of contralateral labeling after caudal motor cortex injections was significantly more extensive for immature than mature animals. In immature animals, most of the labeling was present in laminae 5-7 (dorsal portion). with lesser amounts in laminae 1-4 and 7 (ventral portion), 8, and 9. In mature animals, there were significant reductions in the amount of label in laminae 7-9, resulting in contraction of the labeled territory. The distribution of dorsal horn-ventral horn labeling shifted from 41% and 59% in immature animals to 77% and 23% in maturity. The distribution of contralateral labeling after rostral motor cortex injections also was different in immature and mature animals, but the changes were less extensive than for the caudal motor cortex. In immature animals, the distribution of labeling was similar to that after caudal motor cortex injections in animals of the same age. In mature animals, there was a significant reduction in the amount of labeling in laminae 1-4 and a smaller reduction in 7 (ventral)-9. The overall dorsal horn-ventral horn distribution, however, remained largely unchanged, from 40% and 60% in immature animals to 44% and 56% in maturity. In immature animals after rostral motor cortex injection, corticospinal terminations were present within the lateral motor nuclei. Thus, the distinctive spinal termination patterns of caudal and rostral motor cortex in maturity each reflected postnatal refinement of the distribution of axon terminations. It is plausible that the dorso-ventral refinement of corticospinal terminations relies on activity-dependent competition between caudal and rostral motor cortex, similar to activity-dependent refinement of the laterality of terminations. Our results, however, suggest that the developmental program for achieving corticospinal connectional specificity by the two subregions is different because there was large-scale refinement of the terminations of caudal motor cortex but only local refinement of rostral motor cortex terminations. PMID- 11037286 TI - Rapid functional plasticity of the somatosensory cortex after finger amputation. AB - Recent research indicates that areas of the primary somatosensory (SI) and primary motor cortex show massive cortical reorganization after amputation of the upper arm, forearm or fingers. Most of these studies were carried out months or several years after amputation. In the present study, we describe cortical reorganization of areas in the SI of a patient who underwent amputation of the traumatized middle and ring fingers of his right hand 10 days before cortical magnetic source imaging data were obtained. Somatosensory-evoked magnetic fields (SEF) to mechanical stimuli to the finger tips were recorded and single moving dipoles were calculated using a realistic volume conductor model. Results reveal that the dipoles representing the second and fifth fingers of the affected hand were closer together than the comparable dipoles of the unaffected hand. Our findings demonstrate that neural cell assemblies in SI which formerly represented the right middle and ring fingers of this amputee became reorganized and invaded by neighbouring cell assemblies of the index and little finger of the same hand. These results indicate that functional plasticity occurs within a period of 10 days after amputation. PMID- 11037287 TI - The effect of internal GTPgammaS on GABA-release in cultured hippocampal neurons. AB - The effects of presynaptic guanosine-5'-O-(3-thio)triphosphate (GTPgammaS) on GABAergic inhibitory postsynaptic currents (IPSCs) were studied in cultured hippocampal neurons using whole-cell recordings. Inclusion of GTPgammaS (0.5-1 mM) in the presynaptic electrode reduced both the amplitude and paired-pulse depression of IPSCs, indicating that the probability of GABA-release had been reduced. Presynaptic GTPgammaS increased the depression of IPSCs by the GABA(B) receptor-agonist baclofen (10 microM), and the effect of baclofen was poorly reversible after washing. Stimulation of the GABAergic neuron at 80 Hz for 1 s was accompanied by tetanic depression of the IPSCs by 52+/-6% and was followed by post-tetanic potentiation (PTP), reaching a peak value of 71+/-21% and lasting about 100 s. IPSCs evoked after tetanic stimulation were depressed and PTP was absent when tetanic stimulation was applied within 3 min after starting injection of GTPgammaS into the presynaptic neuron. At longer times, basal release underlying a single IPSC was depressed. This affected the ratios recorded in response to tetanic stimulations such that tetanic depression was abolished, while PTP increased to 117+/-34%. In conclusion, GTPgammaS reduces the probability of GABA-release in both a use- and time-dependent manner, most likely through an inhibitory action on presynaptic Ca2+-influx through voltage-gated Ca2+ channels or an interaction with small GTP-binding proteins in the nerve terminals. PMID- 11037288 TI - Torsional vestibulo-ocular reflex during whole-body oscillation in the upright and the supine position. I. Responses in healthy human subjects. AB - In rhesus monkeys, the dynamic properties of the torsional vestibulo-ocular reflex (VOR) are modified by otolith input: compared with torsional oscillations about an earth-vertical axis (canal-only stimulation), the phase lead observed at frequencies below 0.1 Hz is cancelled when the animals are rotated about an earth horizontal axis (canal-and-otolith stimulation); the gains of the torsional VOR, however, are nearly identical in both conditions. To test whether or not canal otolith interaction in humans is similar to that in rhesus monkeys, we examined ten healthy human subjects on a three-axis servo-controlled motor-driven turntable. The subjects were oscillated in upright or supine position in complete darkness over a similarly wide range of frequencies (0.05-1.0 Hz) with peak velocities <40 degrees/s. Eye movements were recorded using the three-dimensional search coil technique. Compared with the torsional vestibulo-ocular gains during canal-stimulation only (earth-vertical axis), the gains obtained during combined canal-otolith-stimulation (earth-horizontal axis) were significantly higher throughout the entire frequency range (P<0.05). The gain increased by 0.100+/ 0.074 (SD), independent of frequency. During the earth-horizontal axis stimulation, the phase remained always around zero, which is in contrast to the canal-stimulation only, during which one finds an increasing phase lead as frequency decreases. We conclude that, in healthy humans as in rhesus monkeys, the phase lead from the canal signals at low frequencies is effectively cancelled by the otolith input. In contrast to rhesus monkeys, however, otolith signals in healthy humans increase the gain of the torsional VOR at frequencies from 0.05 to 1.0 Hz. This normal database is crucial for the interpretation of results obtained in patients with vestibular disorders. PMID- 11037289 TI - Spatial localization in patients with unilateral posterior left or right hemisphere lesions. AB - In this study, perceptual and visuomotor spatial localization were examined in patients with unilateral right (RH) or left (LH) hemisphere lesions and in a group of control subjects. Perceptual localization was measured with a position discrimination task; in the visuomotor localization task, subjects had to point to a visual target. Both tasks were investigated in conditions with or without background visibility and with central and peripheral targets. In the visuomotor task, hand visibility was also manipulated. In both tasks, targets were presented in the left and right visual hemifield. The perceptual task revealed impairments for both LH and RH patients in the contra-lateral visual hemifield. RH patients also revealed slightly larger impairments in conditions without a visual background. In the visuomotor task, the LH patients were not impaired, whereas the RH patients were impaired in conditions without hand visibility and/or background visibility. Hence, our data strengthen the idea that spatial localization is not a unitary function and that perceptual and visuomotor localization can be selectively impaired. We suggest that one of the important factors distinguishing between localization impairments in RH and LH patients might be absolute versus relative localization. PMID- 11037290 TI - Segment interdependency and difficulty in two-stroke sequences. AB - We previously demonstrated that velocity and movement time for the initial segment for a two-stroke movement are scaled in relation to the difficulty of the second segment. The interdependent kinematic changes were interpreted as evidence that movement planning/organization processes consider the movement parameters of both segments when determining the movement characteristics of the entire sequence. In this experiment we examined two-stroke movements where the difficulty of the first segment had either a low or high level of difficulty to determine if the interdependent kinematic changes are diminished when parameter specification is high for the initial segment. Two-stroke arm movements toward defined targets were made in the horizontal plane on an x-y digitizer. The direction of the first segment was an elbow extension movement away from the trunk. The direction of the second segment varied between forearm extension and flexion movements. Two different indexes of difficulty (IDs) of the first segment and two of the second segment were created by varying target size. In the low ID condition for the first segment, movement duration of the initial segment lengthened and peak velocity decreased when the ID of the second segment was increased, and this pattern was found for both the extension-extension and extension-flexion sequences. In contrast, when the level of difficulty was high for the first segment, the interdependencies disappeared for the extension extension sequence: movement duration and peak velocity were unaffected by the difficulty of the second segment. For the extension-flexion sequence, however, the interdependencies were found in the movement time of the initial segment but were eliminated in the peak velocity, i.e., movement time increased, but the peak velocity did not change. Furthermore, for both the extension-extension and extension-flexion sequences, the intersegment interval was lengthened as the level of difficulty increased. These findings suggest that difficulty of the initial segment affects how the motor planning/organization processes treat adjacent segments of the sequence. In particular, the data support the hypothesis that when the initial movement segment has a high index of difficulty, motor planning/organization processes appear to treat the adjacent segments separately as two discrete actions. PMID- 11037291 TI - Basic response determinants of single neurons to amplitude modulation in the auditory midbrain. AB - Amplitude-modulated (AM) signals represent important components of environmental sounds. While single-cell responses to AM tones in the central auditory system were often studied using repetitive modulation, owing to its presence in vocalization signals, the AM response has not been fully depicted in terms of receptive field in the stimulus domain. This study was aimed to characterize the receptive field of AM response with respect to nonrepetitive AM stimuli and to understand how complex acoustic signals may be coded in the brain. A novel AM stimulus was implemented with a random envelope and a systemic change in intensity across trials. From 393 single units recorded in the inferior colliculus (IC) of urethane-anesthetized rats, responses to the AM stimulus were first characterized in terms of dot-raster pattern. Three types of response were identified: type I showing a monotonic response to mainly the steady states of the AM envelope and type II to rising phases of the AM envelope with a clear intensity preference. Type III showed a mixed response of both type I and type II. A small number of units, called type IV, responded to both rising and falling phases of the modulation. Using perispike averaging, the AM receptive field, or "level temporal receptive field" (LTRF), was displayed in a "stimulus level versus perispike time" plane. The LTRF, particularly of the type II response, clearly revealed triggering features of the cell. The triggering features are consistent with the representation of the cell's response in a receptive space formed by the Cartesian axes of the velocity of amplitude modulation, the intensity of the sound, and the range of modulation. We therefore considered these stimulus parameters as the three basic determinants of the AM response in the auditory midbrain. PMID- 11037292 TI - Acquisition of a new motor skill is accompanied by changes incutaneomuscular reflex responses recorded from finger muscles in man. AB - Cutaneomuscular reflex (CMR) responses and motor unit synchronisation have been recorded to investigate possible reorganisation of central nervous pathways in six healthy adults learning a novel skill with the non-dominant hand. Multi-unit surface EMG signals were recorded from first dorsal interosseous (1DI) and abductor digiti minimi (ADM) muscles during sustained flexion of the index finger and abduction of the little finger (flex/ab) and during bilateral controlled finger abduction (ab/ab). CMR responses were elicited by concomitant stimulation of the digital nerves of the index finger at 2.5 times threshold for perception. For training purposes the subject practised the novel flex/ab task phasically at a rate of 1 Hz for 10 min daily for 12 days. Cortically mediated components of the CMR responses recorded from 1DI and ADM were significantly larger in both muscles during training. Taking the data for all subjects together, when subjects performed sustained flex/ab (novel task), the size of the I1 and E2 components were increased by 25 and 55% in IDI and 192 and 167% in ADM (Mann-Whitney U-test, P<0.05). Corresponding values for sustained ab/ab were more modest: 31 and 16% in 1DI and 88 and 54% in ADM. Changes in the size of the spinal E1 component were not the same in each muscle: values for sustained flex/ab and ab/ab were 33 and 31% smaller in 1DI and 89 and 59% larger in ADM. No significant changes were found in the amount of synchrony of motor unit firing between 1DI and ADM when subjects performed either task during training. These results suggest that learning a new motor skill produces changes which take place predominantly in the cortical pathways of the CMR and these may be due to changed connectivity within motor and/or sensory cortex which has previously been shown in the monkey. PMID- 11037293 TI - Effects of cholinergic neuromodulation in cerebellar flocculus on transparent motion processing in the rabbit. AB - The way a rabbit moves its eyes in response to a stimulus consisting of two moving random dot patterns largely depends on the relative luminances of the two patterns. Concurrent rotation of the animal enhances the response to the visual pattern that represents the same head movement as the vestibular stimulation. In this paper we investigate the role that the flocculus plays in this behaviour. We injected the non-selective acetylcholine agonist carbachol into the flocculus. These injections are known to increase the gain of the optokinetic reflex, but have a smaller effect on the vestibulo-ocular reflex. We investigated the effect on the oculomotor response to (vestibulo-) transparent stimuli, where one pattern oscillated sinusoidally and the other pattern was stable with respect to the head. We found that the injections caused a higher response gain at a lower luminance of the oscillating pattern. Furthermore the influence of concurrent vestibular stimulation decreased. These findings agree with a role of the flocculus that is downstream of the visual normalisation, but upstream of the visual-vestibular interaction. PMID- 11037294 TI - Rapid modifications of somatostatin neuron activity in the periventricular nucleus after acute stress. AB - We have previously reported that stress induces a rapid increase in hypothalamic somatostatin (SS) release. In the present work, we investigated whether SS synthesis is also affected by this treatment. Male rats were subjected to 15-min immobilization (IMO) stress, and measurements of both SS mRNA levels and SS mRNA containing cells were analyzed in the periventricular nucleus (PeV) by radioactive and nonradioactive in situ hybridization (ISH), respectively. In addition, SS content and total SS mRNA were measured in the whole hypothalamus by radioimmunoassay (RIA) and northern blot analysis, respectively. ISH was conducted by applying either a radioactive-labeled (35S) or a digoxigenin (DIG) labeled oligonucleotide probe on histological sections containing the periventricular region of the anterior hypothalamic area (AHA). ISH analysis using radioactive label showed a significant increase in SS mRNA levels in stressed rats. In contrast, stress treatment decreased the number of DIG-labeled cells expressing SS mRNA in this region by 35% as compared to the same histological sections from naive control rats. In addition, a significant decrease in the total SS mRNA DIG-labeled area was observed. Finally, SS content and SS mRNA measured in the whole hypothalamus of stressed rats were markedly inhibited as compared to control rats. Our data show that IMO stress induces a significant and rapid increase in SS mRNA level accompanied by a decrease in the number of cells expressing SS mRNA in the PeV-AHA. The present results suggest that a subset of PeV SS neurons, which became silent at the onset of stress, are regulated independently of the remaining whole mass of PeV neurons. This differential control is in line with the cellular heterogeneity described in periventricular SS-producing neurons and with the multiple hypothalamic and pituitary functions assigned to SS. PMID- 11037295 TI - A test between two hypotheses and a possible third way for the control of prehension. AB - We used an obstacle avoidance task to test two opposing accounts of how the nervous system controls prehension. The visuomotor account supposes that the system independently controls the grip formation and transport phase of prehensile movements. In contrast, the digit channel hypothesis suggests that the system controls the thumb and finger more or less independently. Our data strongly favoured the traditional visuomotor channel hypothesis and demonstrated that the time taken to grasp an object in the presence of obstacles was well predicted by a Fitts' law relationship. We suggest a "thirdway" hypothesis in order to retain the advantages of the digit channel hypothesis within the visuomotor framework. The third-way hypothesis suggests that the nervous system selects a single digit to transport to the object. We speculate that the actual digit selected might depend upon attention and the nature of the prehension task. This hypothesis is able to account for most of the empirical findings unearthed by researchers investigating the control of prehension. PMID- 11037296 TI - Distribution of non-monosynaptic excitation to early and late recruited units in human forearm muscles. AB - The distribution of monosynaptic and nonmonosynaptic excitation was investigated within flexor carpi radialis (FCR) and extensor carpi radialis (ECR) motoneurone (MN) pools. FCR H reflexes of different size were conditioned by various conditioning stimuli eliciting different effects: (1) musculocutaneous-induced non-monosynaptic excitation of FCR MNs at the onset of biceps contraction, (2) heteronymous monosynaptic Ia facilitation, (3) reciprocal Ia inhibition, and (4) presynaptic inhibition of Ia terminals. Musculocutaneous-induced non-monosynaptic excitation increased continuously with the size of the unconditioned reflex. In contrast, heteronymous monosynaptic Ia excitation first increased and then decreased, with increases in the unconditioned reflex size, reciprocal inhibition and presynaptic inhibition showing an approximately similar tendency. This suggests that the non-monosynaptic excitation is distributed more evenly to early and late recruited MNs than monosynaptic Ia excitation, reciprocal inhibition and presynaptic inhibition. A different pattern of homonymous radial-induced monosynaptic and non-monosynaptic excitation was also found for individual ECR MNs investigated with the poststimulus time histogram (PSTH) method. Whereas the monosynaptic Ia excitation tended to be most marked in lower threshold MUs, the nonmonosynaptic excitation was evenly distributed to lower and higher threshold MUs. We propose that the even distribution of the non-monosynaptic excitation in the motoneuronal pool may be of significance when it is necessary to activate a wide range of MNs more or less simultaneously. PMID- 11037297 TI - Recent advances in the molecular basis of inherited skin diseases. AB - Over the last few years the molecular basis of several inherited skin diseases has been delineated. Some discoveries have stemmed from a candidate gene approach using clinical, biochemical, immunohistochemical, and ultrastructural clues, while others have arisen from genetic linkage and positional cloning analyses. Notable advances have included elucidation of specific gene pathology in the major forms of inherited skin fragility, ichthyosis, and keratoderma. These findings have led to a better understanding of the significance of individual structural proteins and regulatory enzymes in keratinocyte adhesion and differentiation. From a clinical perspective, the advances have led to better genetic counseling in many disorders, the development of DNA-based prenatal diagnosis, and a foundation for planning newer forms of treatment, including somatic gene therapy, in selected conditions. PMID- 11037298 TI - Molecular genetics and target site specificity of retroviral integration. AB - Integration is an essential step in the life cycle of retroviruses, resulting in the stable joining of the viral cDNA to the host cell chromosomes. While this critical process makes retroviruses an attractive vector for gene delivery, it also presents a potential hazard. The sites where integration occurs are nonspecific. Therefore,it is possible that integration of retroviral DNA will affect host gene expression and disrupt normal cellular functions. The mechanism by which integration sites are chosen is not well understood, and is influenced by several factors, including DNA sequence and structure, DNA-binding proteins, DNA methylation, and transcription. Integrase, the viral enzyme responsible for catalyzing integration, also plays a key role in controlling the choice of target sites. The integrase domain responsible for target site selection has been mapped to the central core region. A better understanding of the interaction between the target-specifying motif of integrase and the target DNA may allow a means to manipulate integration into particular chromosomal sites. Another approach to directing integration is to fuse integrase with a sequence-specific DNA-binding protein, which results in a bias of integration in vitro into the recognition site of the fusion partner. Successful incorporation of the fusion protein into infectious virions and the identification of optimal proteins that can be fused to integrase will advance the development of site-specific vectors. Retroviruses are promising for the delivery of genes in experimental and therapeutic protocols. A better understanding of integration will aid in the design of safer and more effective gene transfer vectors. PMID- 11037299 TI - Xeroderma pigmentosum and related disorders: defects in DNA repair and transcription. AB - The genetic disorders xeroderma pigmentosum (XP), Cockayne syndrome (CS), and trichothiodystrophy (TTD) are all associated with defects in nucleotide excision repair (NER) of DNA damage. Their clinical features are very different, however, XP being a highly cancer-prone skin disorder, whereas CS and TTD are cancer-free multisystem disorders. All three are genetically complex, with at least eight complementation groups for XP (XP-A to -G and variant), five for CS (CS-A, CS-B, XP-B, XP-D, and XP-G), and three for TTD (XP-B, XP-D, and TTD-A). With the exception of the variant, the products of the XP genes are proteins involved in the different steps of NER, and comprise three damage-recognition proteins, two helicases, and two nucleases. The two helicases, XPB and XPD, are components of the basal transcription factor TFIIH, which has a dual role in NER and initiation of transcription. Different mutations in these genes can affect NER and transcription differentially, and this accounts for the different clinical phenotypes. Mutations resulting in defective repair without affecting transcription result in XP, whereas if transcription is also affected, TTD is the outcome. CS proteins are only involved in transcription-coupled repair, a subpathway of NER in which damage in the transcribed strands of active genes is rapidly and preferentially repaired. Current evidence suggests that they also have an important but not essential role in transcription. The variant form of XP is defective in a novel DNA polymerase, which is able to synthesise DNA past UV damaged sites. PMID- 11037301 TI - Sickness absence with psychiatric disorders--an increased risk for marginalisation among men? AB - BACKGROUND: Sickness absence with psychiatric disorders is a major public health problem with serious consequences for the individual, the employer and society. The aim was to assess the occurrence of psychiatric sickness absence with special focus on sex differences. METHODS: A nationwide sickness insurance register was used. Population at risk was defined as all individuals entitled to sickness benefits in 1994 (N = 1,978,030). Those who were sick-listed for more than 14 consecutive days with a psychiatric diagnosis (n = 28,799) were selected as cases. RESULTS: Of the population under study, 1.46% had at least one psychiatric sickness absence episode. Women had twice the male cumulative incidence of sickness absence for a psychiatric diagnosis. Cumulative incidence was highest among those aged 45-59 years. Men had more sickness absence days. Depression was the most common diagnosis among both women and men. CONCLUSION: Increased efforts are needed to recognise, treat and rehabilitate individuals with a lowered work capacity due to sickness absence. The increased risk of long sick-leave spells among men needs further attention. PMID- 11037300 TI - Primary immunodeficiency mutation databases. AB - Primary immunodeficiencies are intrinsic defects of immune systems. Mutations in a large number of cellular functions can lead to impaired immune responses. More than 80 primary immunodeficiencies are known to date. During the last years genes for several of these disorders have been identified. Here, mutation information for 23 genes affected in 14 immunodefects is presented. The proteins produced are employed in widely diverse functions, such as signal transduction, cell surface receptors, nucleotide metabolism, gene diversification, transcription factors, and phagocytosis. Altogether, the genetic defect of 2,140 families has been determined. Diseases with X-chromosomal origin constitute about 70% of all the cases, presumably due to full penetrance and because the single affected allele causes the phenotype. All types of mutations have been identified; missense mutations are the most common mutation type, and truncation is the most common effect on the protein level. Mutational hotspots in many disorders appear in CPG dinucleotides. The mutation data for the majority of diseases are distributed on the Internet with a special database management system, MUTbase. Despite large numbers of mutations, it has not been possible to make genotype-phenotype correlations for many of the diseases. PMID- 11037302 TI - Mortality in Western Australian psychiatric patients. AB - BACKGROUND: The aim was to examine mortality in psychiatric patients in Western Australia (WA), over a time period of considerable change in the delivery of mental health services. METHODS: A population-based record linkage analysis was undertaken to quantify mortality among people with mental illness in WA. Mortality rates were calculated in users of mental health services and compared with rates in the whole population of WA. Trends in mortality were also examined using relative survival analysis, and proportional hazards regression. RESULTS: The overall mortality rate ratio was 2.57 in males (95% CI: 2.51-2.64), and 2.18 in females (2.12-2.24). The highest cause-specific mortality rate ratio was for deaths due to suicide [RR: 7.37 in males (95% CI: 6.74-8.05) and 8.38 in females (95% CI: 7.11-9.89)], with mortality rate ratios being significantly greater than 1 for all other major causes of death. A relative survival analysis found that the excess mortality risk was concentrated in the first few years after first contact with mental health services. Proportional hazards regression analysis found a slight elevation of mortality rates over time. CONCLUSIONS: Mortality among psychiatric patients remains high and appears to be increasing. Highest excess mortality rate is associated with suicide, but mortality rates are significantly elevated for all major causes of death. PMID- 11037303 TI - Psychometric properties of the schedules for clinical assessment in neuropsychiatry (SCAN-2.1). AB - BACKGROUND: The Schedules for Clinical Assessment in Neuropsychiatry (SCAN), the successor of the ninth version of the Present State Examination (PSE-9), is one of the latest instruments developed by the World Health Organisation for the assessment of psychiatric disorders. So far, the psychometric properties have only been established for certain sections of the instrument. The present study is the first to test the psychometric properties of SCAN-2.1 for most of the disorders covered by the SCAN, and was carried out prior to a survey conducted in the Nijmegen Health Area (the Netherlands). METHODS: Interviewers were psychology graduates with little clinical experience. Two designs were used. In one design, pairs of independent live interviews with the same respondent were compared (test retest situation). In the other, ten videotaped interviews by experts were rated by each of the interviewers (standardized situation), and the outcomes were compared with those of the other interviewers as well as with a reference score. RESULTS: In the test-retest situation the kappa coefficient for diagnostic caseness was qualified as substantial (0.62) and for diagnostic categories and diagnostic groups as moderate to good (0.24 to 0.64). In the standardized situation using videotaped interviews by experts, sensitivity as well as specificity proved to be substantial to almost perfect. The agreement per interviewer with regard to the reference diagnoses ranged from 87% (diagnostic group) to 94% (diagnostic caseness). Agreement on the syndrome level (without duration and interference criteria of DSM-IV) was excellent. CONCLUSIONS: Although the instrument is traditionally used by experienced clinicians, this study shows that less experienced (but well trained) interviewers can apply SCAN reliably. Special attention should be paid to the items without explicit interview questions, as they tend to be more sensitive to neglect than the items with interview questions. PMID- 11037304 TI - The Parental Bonding Instrument: confirmatory evidence for a three-factor model in a psychiatric clinical sample and in the National Comorbidity Survey. AB - BACKGROUND: Study of the contribution of retrospective perceptions of dysfunctional parenting in relation to adult psychopathology has been greatly facilitated by the development of the 25-item Parental Bonding Instrument (PBI; Parker et al. 1979). METHOD: The present study employed confirmatory factor analytic techniques to evaluate competing models of the basic dimensions underlying different versions of the PBI, in a psychiatric sample from a mood disorders program and with a new modification of the PBI employed in the US National Comorbidity Survey. RESULTS: The results indicated that a three-factor model originally identified in a 16-item version of the PBI modified for epidemiological research (Kendler 1996) showed the best fit to the data. The three dimensions of care, overprotection, and authoritarianism also explained the underlying structure of the NCS-modified, eight-item PBI that is now part of the NCS public use dataset available to psychopathology researchers. CONCLUSIONS: The replicability of findings across gender, age, and clinical versus community samples attests to the robustness of this three-factor structure of parenting styles. PMID- 11037305 TI - Marital and family functioning: different measures and viewpoints. AB - BACKGROUND: Dysfunctional relationships have been considered to play an important part in the onset and maintenance of psychiatric disorders, particularly depression. Influential factors appear to be perception of low social support and emotional warmth and/or high levels of criticism and control by the recipient. The IBM was developed for use as a simple self-report measure to rate important components of the relationship between marital partners, described as the constructs of care and control. The aim of this study was to test the validity of the IBM. METHODS: The IBM was compared with data from a structured marital interview based on the Self-Evaluation of Social Support Schedule (SESS). Data were also obtained from a nominated adult offspring witness. Family functioning was assessed using the General Functioning component of the McMaster Family Assessment Device (FAD). The General Health Questionnaire (GHQ) was used to determine levels of psychological morbidity. The witness data for sons and daughters were analysed to gauge gender effects in reporting. RESULTS: The results show that 'care' and 'control' were identifiable constructs discerned by partners and witnesses with 'care' rated more consistently than 'control'. Care between parents was an indicator of the overall quality of the family environment. Care received by the wife from her husband seemed to set the emotional tone for the family. Daughters seemed to be more 'in tune' with perceived care by both parents. Sons were less so, overall, but were more 'in tune' with their fathers' perceived care than with their mothers'. CONCLUSION: Both the IBM, a self-report measure, and the structured marital interview provided consistent information about the quality of marital relationships, particularly perceptions of care. Perceptions of control were less consistently reported, which may suggest that 'control' is a less robust construct. This may suggest that 'control' is a more subjective experience, as the cues are more directed at the partner than other family members. PMID- 11037306 TI - Social support, personality and depressive symptoms over 7 years: the Health and Lifestyle cohort. AB - BACKGROUND: We tested the hypothesis that a negative relationship between social support and depression is stronger in extroverts. METHODS: Data on social support and personality were obtained from an existing cohort of 9003 adults (the Health and Lifestyle Survey, UK), of whom 3594 respondents who were followed-up 7 years later contributed to the present analysis. Six depression items from the 30-item General Health Questionnaire, summed, were divided into five levels and a proportional odds analysis was performed. Information on social support was also obtained at follow-up. RESULTS: For females, there was a highly significant interaction between Time of Residence in Area and extroversion (P<0.001). For males, interactions involving Adults in Household and Living as Married reached borderline significance (0.050.10). CONCLUSION: It is unlikely that a lack of social support is more or less harmful for introverts, although the hypothesis should be re-tested in a study with a much shorter period of follow-up and higher response rate. PMID- 11037307 TI - Application of stratum-specific likelihood ratios in mental health screening. AB - BACKGROUND: The accuracy of a diagnostic procedure is commonly assessed by measuring sensitivity, specificity and positive and negative predictive values. Likelihood ratios provide an alternative method for describing these results, though they are typically reported only for dichotomized outcomes. However, likelihood ratios can also be applied to ordinal or continuous results. METHODS: The present paper discusses the application of stratum-specific likelihood ratios in a primary care setting using the General Health Questionnaire (GHQ-12) and the Symptom Check List 90-R (SCL-90-R). A randomly selected sample (n = 408) of adult outpatients from primary care offices in Dusseldorf was screened using the German versions of the GHQ-12 and the SCL-90-R. RESULTS: Logistic regression analysis indicated that stratum-specific or multilevel likelihood ratios preserve more information than a fixed threshold approach with a single cutoff point. For each test, five clinically useful strata with monotonically increasing stratum specific likelihood ratios were selected. CONCLUSIONS: Stratum-specific likelihood ratios have enormous practical value, and they are becoming an important way of expressing and comparing the usefulness of different tests. Stratum-specific likelihood ratios reduce the spectrum bias that might arise if only two categories (cases and non-cases) are chosen. Additionally, multilevel likelihood ratios can be used as bedside information to obtain the post-test probability from the pre-test probability of the disorder. PMID- 11037308 TI - Mortality among persons with a history as psychiatric inpatients with functional psychosis. AB - BACKGROUND: An important aim in all psychiatric care should be a reduction of overall mortality. Information on mortality patterns in different types of psychiatric populations is vital for a successful design of treatment strategies and preventive programmes. The present study aims to describe mortality among persons with a history as psychiatric inpatients with functional psychosis. METHODS: All psychiatric inpatients, 17,878 men and 23,256 women, registered in the Swedish National Hospital Discharge Registry between 1978 and 1982 with a functional psychosis (ICD-8 = 295-299) as principal diagnosis were followed for mortality during the time period 1983-85. Life tables were constructed and death rates for various types of causes of death were calculated. RESULTS: Compared to the general population, the excess mortality in the study group caused a reduction in life expectancy of 22.1-27.9% (95% CI) among the men and 15.0-21.7% among the women. In the age group 2049 years, 62% of the excess mortality was caused by suicide. In the age group 50-89 years, only 8% of the excess was suicide, while 52% was caused by cardiovascular disorders. CONCLUSIONS: Reduction in mortality rates requires different strategies in different age groups. Even if the suicide rate were reduced to zero, it would only have a marginal influence on the highly elevated mortality among patients in upper middle age and among the elderly. Among the younger patients (<35 years), on the other hand, practically all excess mortality was caused by suicide and accidents. PMID- 11037309 TI - Newton Morton: the Wisconsin years. PMID- 11037310 TI - Newton Morton's influence on genetics: the Morton number. PMID- 11037311 TI - Genetic dissection of complex traits: an overview. AB - Genetic dissection of even simple Mendelian traits has been sufficiently challenging. Complex traits are proving to be much more challenging and frustrating than previously thought. The concepts, methods, and strategies discussed in this volume emphasize the critical importance of study design, appropriate methods of analysis, including relatively newer and emerging methods, and issues relating to the interpretation of results from genome scans; some thoughts on the future the new millennium holds are offered, as well. This chapter overviews the key steps involved in the study of complex traits, which are discussed in detail in subsequent chapters. It is suggested that a combination of lumping and splitting strategies is more appropriate for the analysis of complex traits, and large-scale collaborations should make this possible. For example, by pooling data and/or results from multiple studies on a given disease/trait, one may attain a sample size large enough to permit the division of the data into multiple relatively more homogeneous subgroups. The sample size of the subgroups may still be sufficiently large sample, but the genetic dissection within each subgroup should be much less daunting. The expectation is that analyses within subgroups will enhance gene finding, especially when any interacting determinants are taken into account at the time of dividing the data into subgroups. Perhaps the methods are not yet optimum, but the future holds much promise. In the meantime, the cutting-edge methods discussed in this volume by leading experts should help. There is an increasing healthy tendency for investigators to collaborate by pooling materials and results across studies, with the goal of increasing the sample size and thus the power. We believe that such efforts are essential for the genetic dissection of complex traits and should contribute to greater success, especially if there is a real commitment to meaningful collaboration. After all, for most complex traits, the question is not whether there are genes, only when and how they might be found. PMID- 11037312 TI - Familial resemblance and heritability. AB - Familial resemblance, which arises when members within families are more similar than are unrelated pairs of individuals, may be estimated in terms of correlations (or covariances) among family members. The magnitudes of such correlations generally reflect both the extent of environmental sharing and the degree of biological relationship between the relatives. Heritability, or more appropriately multifactorial heritability or generalized heritability, quantifies the strength of the familial resemblance and represents the percentage of variance ina trait that is due to all additive familial effects including additive genetic effects and those of the familial environment. However, the traditional concept of heritability, which may be more appropriately called the genetic heritability, represents only the percentage of phenotypic variance due to additive genetic effects. Resolving the sources of familial resemblance entails other issues. For example, there may be major gene effects that may be largely or entirely nonadditive, temporal or developmental trends, and gene-gene (epistasis) and gene-environment interactions. The design of a family study determines which of these sources are resolvable. For example, in intact nuclear families consisting of parents and offspring, the genetic and familial environmental effects are not resolvable because these relatives share both genes and environments. However, extended pedigrees and twin and adoption study designs allow separation of the heritable effects and, possibly, more complex etiologies, including interactions. Various factors affect the estimation and interpretability of heritability, for example, assumptions regarding linearity and additivity of the effects, assortative mating, and the underlying distribution of the data. Nonnormality of the data can lead to errors in hypothesis testing, although it yields reasonably unbiased estimates. Fortunately, these and other complications can be directly modeled in many of the sophisticated software packages available today in genetic epidemiology. PMID- 11037313 TI - Linkage and association: basic concepts. AB - Many investigators are turning their efforts to dissecting the etiology of complex traits. The primary tools for gene discovery, localization, and functional analysis are linkage and association studies. While the conceptual underpinnings of these approaches have long been known, advances in recent decades in molecular genetics, in the development of efficient computational algorithms, and in computing power have enabled the large-scale application of these methods. Here, we review the biological basis of linkage and association among loci and the common methods used to assess these relationships with respect to observed phenotypes. We further consider the two most common approaches- genome scans and candidate gene studies--especially their respective strengths, weaknesses, and resource requirements. Finally, we highlight some of the major challenges that arise from these investigative approaches and those that are inherent in the nature of complex traits. The chapters that follow elaborate on many of these topics. PMID- 11037314 TI - Definition of the phenotype. AB - Definition of the phenotype is a key issue in designing any genetic study whose goal is to detect disease genes. This chapter describes strategies to increase the power to detect susceptibility loci for complex diseases. A narrowly defined disease phenotype can offer advantages over broad definitions. Studies of clinical disease can also benefit from judicious selection of endophenotypes and related quantitative traits for analysis. The effect of diagnostic and measurement error is also discussed; power is maximized when strategies to reduce error are incorporated into a study design. PMID- 11037315 TI - Genotyping for human whole-genome scans: past, present, and future. AB - Efficient and effective whole-genome 10-cM short tandem repeat polymorphism (STRP) scans are now available. Doubling or tripling STRP density to an average spacing of 3-5 cM is readily achievable. However, if typing costs for diallelic polymorphisms can be brought close to, or preferably less than, one-third those of STRPs, then diallelics may gradually supplement or supplant STRPs in whole genome scans. The power of higher density genome scans for gene map ping by association and for many other research and clinical applications is great. It would be wise to continue investing heavily for many years in genotyping technology. PMID- 11037316 TI - The lod score method. AB - The lod score method originated in a seminal article by Newton Morton in 1955. The method is broadly concerned with issues of power and the posterior probability of linkage, ensuring that a reported linkage has a high probability of being a true linkage. In addition, the method is sequential, so that pedigrees or lod curves may be combined from published reports to pool data for analysis. This approach has been remarkably successful for 50 years in identifying disease genes for Mendelian disorders. After discussing these issues, we consider the situation for complex disorders, where the maximum lod score (MLS) statistic shares some of the advantages of the traditional lod score approach but is limited by unknown power and the lack of sharing of the primary data needed to optimally combine analytic results. We may still learn from the lod score method as we explore new methods in molecular biology and genetic analysis to utilize the complete human DNA sequence and the cataloging of all human genes. PMID- 11037317 TI - Extension of the lod score: the mod score. AB - In 1955 Morton proposed the lod score method both for testing linkage between loci and for estimating the recombination fraction between them. If a disease is controlled by a gene at one of these loci, the lod score computation requires the prior specification of an underlying model that assigns the probabilities of genotypes from the observed phenotypes. To address the case of linkage studies for diseases with unknown mode of inheritance, we suggested (Clerget-Darpoux et al., 1986) extending the lod score function to a so-called mod score function. In this function, the variables are both the recombination fraction and the disease model parameters. Maximizing the mod score function over all these parameters amounts to maximizing the probability of marker data conditional on the disease status. Under the absence of linkage, the mod score conforms to a chi-square distribution, with extra degrees of freedom in comparison to the lod score function (MacLean et al., 1993). The mod score is asymptotically maximum for the true disease model (Clerget-Darpoux and Bonaiti-Pellie, 1992; Hodge and Elston, 1994). Consequently, the power to detect linkage through mod score will be highest when the space of models where the maximization is performed includes the true model. On the other hand, one must avoid overparametrization of the model space. For example, when the approach is applied to affected sibpairs, only two constrained disease model parameters should be used (Knapp et al., 1994) for the mod score maximization. It is also important to emphasize the existence of a strong correlation between the disease gene location and the disease model. Consequently, there is poor resolution of the location of the susceptibility locus when the disease model at this locus is unknown. Of course, this is true regardless of the statistics used. The mod score may also be applied in a candidate gene strategy to model the potential effect of this gene in the disease. Since, however, it ignores the information provided both by disease segregation and by linkage disequilibrium between the marker alleles and the functional disease alleles, its power of discrimination between genetic models is weak. The MASC method (Clerget-Darpoux et al., 1988) has been designed to address more efficiently the objectives of a candidate gene approach. PMID- 11037318 TI - Major strengths and weaknesses of the lod score method. AB - Strengths and weaknesses of the lod score method for human genetic linkage analysis are discussed. The main weakness is its requirement for the specification of a detailed inheritance model for the trait. Various strengths are identified. For example, the lod score (likelihood) method has optimality properties when the trait to be studied is known to follow a Mendelian mode of inheritance. The ELOD is a useful measure for information content of the data. The lod score method can emulate various "nonparametric" methods, and this emulation is equivalent to the nonparametric methods. Finally, the possibility of building errors into the analysis will prove to be essential for the large amount of linkage and disequilibrium data expected in the near future. PMID- 11037319 TI - Overview of model-free methods for linkage analysis. AB - Methods of model-free linkage analysis do not require a detailed specification for the mode of inheritance of the trait locus being linked. Beginning with methods proposed by Penrose in the 1930s, which allowed detection of linkage only, these methods now allow one to use multipoint analysis both to locate trait genes and to estimate variance components that give information on the genetic mechanism underlying the trait. The newer methods can utilize data on multiple types of pairs of relatives other than just sibpairs, and they can detect multiple trait loci. In combination with special sampling schemes, these methods give hope that they may play a crucial role in unraveling the genetic etiology of multifactorial traits, regardless of whether epistatic interactions are present. The results of such analyses can guide the use of more powerful model-based linkage analyses. PMID- 11037320 TI - Variance component methods for detecting complex trait loci. AB - Variance component-based linkage analysis has become a major statistical tool for the localization and evaluation of quantitative trait loci influencing complex phenotypes. The variance component approach has many benefits--it can, for example, be used to analyze large pedigrees, and it is able to accommodate multiple loci simultaneously in a true oligogenic model. Important biological phenomena such as genotype-environment interaction and epistasis are also examined easily in a variance component framework. In this chapter, we review the basic statistical features of variance component linkage analysis, with an emphasis on its power and robustness to distributional violations. PMID- 11037321 TI - Linkage and association with structural relationships. AB - The use of structural equations (path analysis) provides an alternative, equivalent formulation to variance components models. Instead of partitioning the variance, we focus on modeling the underlying random variables themselves through a system of linear, mixed model, regression equations. A few specific examples of genetic path models for linkage and association (linkage disequilibrium) are discussed. This formulation provides a simple yet elegant framework that can continue to be extended to meet the challenges of modeling and dissecting the genetic nature of complex traits in the new century. PMID- 11037322 TI - The future of genetic case-control studies. AB - The case-control study design has been a veritable workhorse in epidemiological research since its inception and acceptance as a valid and valued field of inquiry. The reasons for this owe to the simplicity of the required sampling and the (potential) ease of analysis and interpretation of results. Unfortunately, there are a number of problems that plague the use of the case-control design in assessing relationships between genetic variation and disease susceptibility in the population at large. Many of these problems are entirely analogous to problems that inhere in applications of the case-control design in nongenetic settings. These problems include stratification, the assessment of statistical significance, heterogeneity, and the interpretation of multiple outcomes or phenotypic information. In this chapter we describe 10 problems thought to plague genetic case-control studies and offer potential solutions to each. Many of our proposed solutions require the use of multiple DNA markers to accommodate the genetic background of the individuals sampled as cases and controls. It is hoped that our discussions and proposals will spark further debate about the analysis and ultimate utility of the case-control study in genetic epidemiology research. PMID- 11037323 TI - Cost of linkage versus association methods. AB - Identifying genetic factors that influence disease risk is a major goal in genetic epidemiology. In the past, for disease with relatively simple etiology, genetic linkage methods have been highly effective for this purpose. However, as we begin to study more complex diseases and disorders for which each specific genetic factor may play a minor role in causation, the relative values of genetic linkage methods that do not require linkage disequilibrium versus association based methods that do require linkage disequilibrium must be evaluated. Here, we compare the cost-effectiveness of linkage and association methods for identifying genetic factors for a quantitative trait locus that explains 10% of interindividual variability. We find that the choice of analytical scheme depends upon the degree of disequilibrium in the population. Because this parameter has not been adequately assessed, planning association studies is currently difficult. PMID- 11037324 TI - Genotype-environment interaction in transmission disequilibrium tests. AB - Transmission disequilibrium tests (TDTs) provide an approach to the detection of associations between alleles at marker loci and risk of complex disorders. The logistic regression approach to TDTs proposed by Sham and Curtis (1995) is generalized to provide separate tests of the main effects of marker loci on genetic risk and genotype-environment interaction (G x E) arising because multiple alleles differ in their sensitivity to specified environmental covariates. A modification of the same model may be used to detect the effects of genomic imprinting on the expression of susceptibility loci. In the presence of G x E, highly significant genetic effects may be present that will not produce marked twin or sibling resemblance and will not yield significant associations in conventional TDTs. However, simulation studies show how the logistic regression model can be used to detect the main effects of marker alleles and their interaction with covariates on continuous outcomes in offspring-parent trios, pairs of siblings and their parents, and monozygotic twin pairs and their parents. TDT tests with MZ with twin pairs permit the detection of alleles whose primary effects on the phenotype are mediated through the control of sensitivity to latent features of the within-family environment. It is shown that although the genotype-environment correlation caused by the environmental effects of parental alleles on offspring phenotypes can produce spurious marker-phenotype association in population studies, the outcome of TDTs is not biased thereby. PMID- 11037325 TI - Major strengths and weaknesses of model-free methods. AB - This chapter discusses some of the principal advantages and disadvantages inherent in the use of model-free (MF) methods. The principal advantage is that one does not need to specify, a priori, a genetic model for the trait of interest, which often is not known for many complex phenotypes of interest. On the other hand, as with all nonparametric approaches, use of model-free methods results in reduced power for detection of linkage compared with model-based methods when the model is correctly specified. The MF methods also have a potential for computational simplicity and are ideally suited for analysis of specific relative sets such as affected sibpairs. The MF methods are ideally suited to the analysis of quantitative traits for which finding and implementing a suitable genetic model for use in a parametric linkage analysis may be cumbersome. On the other hand, for discrete traits, most model-free methods allow for only a simple definition of "affected," making it difficult to consider such factors as age at onset, diagnostic accuracy of phenotype, or sex-specific disease risks. A factor that can be viewed as both a strength and weakness of MF methods is the large number of statistical approaches and implementation options of model-free methods; while providing a number of choices for the more sophisticated users, such variety also may lead to the risk of overanalysis of the data by selecting the approach that gives the desired result. In the end, the choice between model-free and model-based methods will largely depend on the nature of the phenotype under study and the existing knowledge base about its underlying mode of inheritance. PMID- 11037326 TI - Meta-analysis for model-free methods. AB - The intricate nature of complex genetic traits dictates that novel methodologies be developed and utilized to achieve better power, better accuracy, and more favorable balance between type I and type II errors than could be achieved by the traditional methods as they are used in mapping Mendelian traits. Meta-analysis provides one such method for synthesizing information from multiple studies. This has the advantage of being able to pool relatively weak signals from individual studies into a collectively stronger evidence of genetic effects, while at the same time providing a quantitative framework for modeling variability among studies. The traditional lod score measures significance level of a linkage effect in an individual study, and its additive property make it a natural candidate for combining results across independent studies. To incorporate the within-study variation of the linkage effect into the pooled overall measure of genetic effect, the effect sizes (such as the proportion of genes shared identical-by-descent, IBD) should be pooled directly across studies. Traditional regression models and mixed effects models can be used to estimate the overall genetic effect size and its variance, and to test heterogeneity among studies. Our simulation studies show that designing studies with moderate power and pooling their results via meta-analysis may be more cost-effective than large dedicated studies. We believe that, as a newly emerging methodology, the meta analysis approach has the potential to become an integral part of our toolbox that will expedite the search for complex human disease genes. PMID- 11037327 TI - Classification methods for confronting heterogeneity. AB - Recursive partitioning/tree models are discussed as a method of dissecting the complex nature of traits with different causal mechanisms operating in different subsets of the data (e.g., different genes operating in different subsets of families). In addition to the straightforward application of classification and regression trees to define more homogeneous subsets of the data on which to conduct further analysis, developments incorporating linkage analysis into the definition of the regression trees (Shannon et al., 2000) are discussed. The pros and cons of recursive partitioning vs. the related approach of context-dependent analysis (Turner et al., 1999) are also reviewed as two promising analysis strategies that may be useful for genetic dissection of complex traits. PMID- 11037328 TI - Applications of neural networks for gene finding. AB - A basic description of artificial neural networks is given and applications of neural nets to problems in human gene mapping are discussed. Specifically, three data types are considered: (1) affected sibpair data for nonparametric linkage analysis, (2) case-control data for disequilibrium analysis based on genetic markers, and (3) family data with trait and marker phenotypes and possibly environmental effects. PMID- 11037329 TI - Genome partitioning and whole-genome analysis. AB - Standard DNA marker-based approaches to mapping genes that influence complex traits typically consider a limited number of hypotheses. Most of these hypotheses concentrate on the effect of a single individual locus (or relatively few loci) on the trait of interest. Although of tremendous importance scientifically, such hypotheses do not accommodate the full range of genetic phenomena that may contribute to phenotypic expression. We present novel approaches to complex trait analysis that make as complete use of marker information as is possible. The proposed methodologies can be used to entertain a wide variety of hypotheses, including those that engage, for example, the contribution of a particular chromosome, genome-wide heterozygosity, and multiple genomic regions, to phenotypic expression. We consider a number of possible extensions of the proposed methods as well as their limitations. Although we discuss many methodological details in the context of quantitative trait locus mapping involving sampling units such as human pedigrees and hybrids resulting from crosses between inbred strains of model organisms, our procedures can be easily adapted to standard sibpair and other sampling unit-based designs. Ultimately, the proposed approaches not only have the potential to increase power to identify individual loci that harbor trait-influencing genes, but also present a framework for testing a number of hypotheses about the nature of the genetic determinants of phenotypes in general. PMID- 11037330 TI - Deciphering the genetic architecture of a multivariate phenotype. AB - A heritable multivariate quantitative phenotype comprises several correlated component phenotypes that are usually pleiotropically controlled by a set of major loci and environmental factors. One approach to decipher the genetic architecture of a multivariate phenotype, in particular to map the underlying loci, is to reduce the dimensionality of the data by means of a data reduction technique, such as principal component analysis. The extracted principal components are then analyzed in conjunction with marker data to map the underlying loci. We have examined the efficiency of this approach with and without taking into account the correlation structure of the multivariate phenotype when extracting principal components. We have assumed that genome-wide scan data on sibpairs are available for low-density (widely spaced) and high density markers. Using extensive simulations, based on three models of the multivariate phenotype, we have shown that although ignoring the correlation structure of the multivariate phenotype does not have any serious impact on the efficiency of mapping the underlying trait loci in wide marker intervals, there is a significant adverse effect of this practice for fine-mapping. We, therefore, recommend that the correlation structure of the multivariate phenotype be carefully examined to decide on the strategy of extracting principal components for deciphering the genetic architecture of the multivariate phenotype. PMID- 11037331 TI - On the resolution and feasibility of genome scanning approaches. AB - Before contemplating a genome scan to identify the map position of disease predisposing genes, an investigator should have prior evidence of the genes' existence. It is therefore logically consistent to evaluate a genome scan experiment as an estimation problem, rather than as a hypothesis-testing problem, since absent prior evidence of the existence of disease genes, it is probably unwise to conduct the experiment at all. Recombination in a single meiosis can be modeled as a point process along the chromosome, and linkage or linkage disequilibrium (LD) mapping statistics are a simple function of the superposition of the recombination processes occurring in all meioses under study. Thus, multipoint lod scores are shown to be step functions, in the absence of ambiguity about the inheritance of chromosomal segments. The ability to map a disease gene is a function of how well the ascertained phenotypes predict the underlying trait locus genotypes. This chapter presents a thorough investigation of the properties of the multipoint lod score and uses results from renewal theory to examine the effects of deviations from a deterministic phenotype-genotype relationship. The quality of estimated gene locations is assessed through computing the mean and variance of the length of the expected 3-lod-unit support interval around the maximum likelihood estimate. The more deterministic the model, the smaller this interval is. A more exact quantification of details of this effect is used to describe the statistical properties of such genome scanning experiments from the perspective of estimation, with appropriately little regard to hypothesis testing. Hypothesis testing, however, is discussed as an appropriate context to describe linkage and LD analysis in situations where candidate genes are being screened, since only there does one have definable null and alternative hypotheses that have not been rejected before the beginning of the experiment. By contrast, it is hoped that the null hypothesis "there is no gene affecting this phenotype" has been rejected by other means before an expensive genome scan is even contemplated (though that this is often not done is probably the main problem!). PMID- 11037332 TI - The role of interacting determinants in the localization of genes. AB - We describe the potential gains in power for localizing disease genes that can be obtained by allowing for interactions with environmental agents or other genes. The focus is on linkage and association methods in nuclear families with dichotomous phenotypes. A logistic model incorporating various main effects and interactions is used for penetrance, but similar methods apply to censored age-at onset or continuous phenotypes. We begin by discussing the influence of gene environment interactions in segregation analysis, illustrated with analysis of smoking as a modifying factor for lung cancer. We then discuss a number of approaches to linkage analysis-model-free and model-based(including generalized estimating equations) incorporating interactions with environmental factors and other genes, either candidate genes or linked loci. We find that a test of heterogeneity in IBD sharing probabilities across strata defined by sharing of environmental factors can offer greater power for detecting linkage than the simple mean test, provided the interaction effect is sufficiently strong; we explore the conditions under which this gain in power occurs. Finally, we describe approaches for testing association and disequilibrium involving interactions, utilizing case-control, case-parent, and pedigree-based approaches. A technical problem that must be addressed in many analyses is the effect of missing data on environmental covariates; we use multiple imputation in an analysis of lung cancer segregation to illustrate an approach to this problem. PMID- 11037333 TI - Linkage disequilibrium mapping: the role of population history, size, and structure. AB - Linkage disequilibrium mapping attempts to infer the location of a disease gene from observed associations between marker alleles and disease phenotype. This approach can be quite powerful when disease chromosomes are descended from a single founder mutation and the markers considered are tightly linked to the disease locus. The success of linkage disequilibrium map ping in fine-scale localization has led to the suggestion that genome-wide association testing might be useful in the detection of susceptibility genes for complex traits. Such studies would likely be performed in small, relatively isolated founder populations, where heterogeneity of the disease is less likely. To interpret the patterns of association observed in such populations, we need to understand the effect of population size, history, and structure on linkage disequilibrium. In this chapter, we first review measures of allelic association at a single locus. Measures of association between two loci are described, and some theoretical results are reviewed. We then consider some methods for inferring linkage between a marker and a rare disease, focusing on those that model the ancestry of the disease chromosomes. Next we discuss factors whose effect on disequilibrium are understood, and finally we describe the characteristics of some human populations that may be useful for disequilibrium mapping of complex traits. PMID- 11037334 TI - Optimum study designs. AB - Because simplistic designs will lead to prohibitively large sample sizes, the optimization of genetic study designs is critical for successfully mapping genes for complex diseases. Creative designs are necessary for detecting and amplifying the usually weak signals for complex traits. Two important outcomes of a study design--power and resolution--are implicitly tied together by the principle of uncertainty. Overemphasis on either one may lead to suboptimal designs. To achieve optimality for a particular study, therefore, practical measures such as cost-effectiveness must be used to strike a balance between power and resolution. In this light, the myriad of factors involved in study design can be checked for their effects on the ultimate outcomes, and the popular existing designs can be sorted into building blocks that may be useful for particular situations. It is hoped that imaginative construction of novel designs using such building blocks will lead to enhanced efficiency in finding genes for complex human traits. PMID- 11037335 TI - One-stage versus two-stage strategies for genome scans. AB - One way to determine the genetic etiology of a complex disease is to find the chromosomal regions that tend to be shared among affected relatives and yet tend to differ between affected and unaffected relatives. This can be done by using equally spaced markers to perform a global linkage search of the whole genome. With the rapid development of molecular technology to readily type individuals, it is becoming relatively easy to carry out such genome searches for disease genes. The simplest approach is to type every individual in the sample at every marker locus. However, such an approach wastes a great deal of effort in genotyping large areas of the genome that are eventually found not to show any evidence for linkage. Efficient design of genome scans for linkage analysis is therefore an important issue. We first describe strategies in which a single sample is typed for markers in one or two stages, with neither the use of a second sample nor any attempt at replication of a linkage result. Then in the discussion we briefly mention other multistage strategies, including one that involves a degree of replication. PMID- 11037336 TI - Significance levels in genome scans. AB - Genome-wide linkage scans using affected sibpair families are being conducted on many complex diseases, such as type 1 and type 2 diabetes, multiple sclerosis, rheumatoid arthritis, schizophrenia, asthma, cardiovascular diseases, obesity, and alcoholism. Despite extensive efforts by many groups, progress has been exceedingly slow, and only a few genes and some genomic regions involved in complex diseases have been identified. The general picture is one of difficulty in locating disease genes and replication of reported linkages. This results from the fact that complex diseases and traits may result principally from genetic variation that is relatively common in the general population involving a large number of genes, environmental factors, and their interactions. Genome-wide association studies are now feasible through the use of PCR methodologies with pooled DNA samples and microsatellite variation, and more recently single nucleotide polymorphism (SNP) variation. Issues relating to significance levels in genome-wide linkage and association scans are discussed, and suggestions for dealing with false positive (type I) errors proposed. PMID- 11037337 TI - False positives and false negatives in genome scans. AB - It is emphasized that two types of errors are made in the testing of a hypothesis, false positive (type I) and false negative (type II). Genome-wide scans involving many markers give rise to the problem of multiple testing, which results in an increased number of false positives, thus necessitating a correction in the nominal significance level. While the literature has concentrated reasonably heavily on controlling false positives in genomic scans, the need to control false negatives has been largely neglected. This chapter highlights this need and attempts to strike a balance between the two error types. The need to develop alternative methods for discriminating between false positives and true positives is also stressed. PMID- 11037338 TI - Sequential methods of analysis for genome scans. AB - As the preceding chapters illustrate, now that whole-genome scan analyses are becoming more common, there is considerable disagreement about the best way to balance between false positives and false negatives (traditionally called type I and type II errors in the statistical parlance). Type I and type II errors can be simultaneously controlled, if we are willing to let the sample size of analysis vary. This is the secret that Wald (1947) discovered in the 1940s that led to the theory of sequential sampling and was the inspiration for Newton Morton in developing the lod score method. We can exploit this idea further and capitalize on an old, but nearly forgotten theory: sequential multiple decision procedures (SMDP) (Bechhoffer, et al., 1968), which generalizes the standard "two hypotheses" tests to consider multiple alternative hypotheses. Using this theory, we can develop a single, genome-wide test that simultaneously partitions all markers into "signal" and "noise" groups, with tight control over both type I and type II errors (Province, 2000). Conceiving this approach as an analysis tool for fixed sample designs (instead of a true sequential sampling scheme), we can let the data decide at which point we should move from the hypothesis generation phase of a genome scan (where multiple comparisons make the interpretation of p values and significance levels difficult and controversial), to a true hypothesis testing phase (where the problem of multiple comparisons has been all but eliminated so that p values may be accepted at face value). PMID- 11037339 TI - From genetics to mechanism of disease liability. AB - The molecular basis of single-gene Mendelian disorders resulting from gain or loss of function is being clarified at a rapid pace. Progress in the genetics of common disease, by contrast, has been frustratingly limited, as we discuss by reference to essential hypertension (EH). The application of standard genetic paradigms to hypertension research has yielded remarkable findings. Arterial pressure (AP) variation in laboratory rats has been correlated with various genes. Likewise, rare Mendelian hypertension syndromes are increasingly understood in molecular terms. The implications of these findings for EH have proven to be modest, however. Genetic methods have been applied to investigate directly essential hypertension in humans, with mixed results. The power of such methods to identify genetic determinants of EH has been questioned. The issues confronting the genetic analysis of EH are discussed by drawing from our ongoing work along the hypothesis that molecular variants of the angiotensinogen gene may constitute inherited predispositions to the condition. Simply establishing correlation is already a daunting task. Far more challenging yet is to establish causation for a physiological phenotype, that is, to understand the mechanism by which a genetic factor may predispose to essential hypertension. Susceptibility imparted by genetic variation, modest and quantitative, modulates response to environmental exposure over time. The product of the gene under examination may be highly pleiotropic, being involved with multiple physiological processes in multiple tissues. Finally, as physiological phenotypes are defined at the level of the entire organism, ultimate demonstration of genetic determination may require specific genetic manipulations in entire organisms. PMID- 11037340 TI - Complex inheritance: the 21st century. AB - At least for the early years of the twenty-first century we can anticipate some of the advances to be made in mapping, positional cloning, pooling of evidence over samples for linkage and allelic association, and fully parametric methods that combine the latter with segregation analysis. This preoccupation with problems the twentieth century failed to solve is not grounds for pessimism if the new century provides solutions and applies them to problems of biological interest. We may hope that genetic epidemiology will be part of a community that addresses the needs of geneticists for international communication, a stable nomenclature, genome databases, and a consensus on ethical, legal, and social issues transcending regional prejudices. PMID- 11037341 TI - Association of human papillomavirus infection with carcinoma of the cervix uteri and its precursor lesions: theoretical and practical implications. AB - Human papillomaviruses (HPVs) are the major aetiological agents of cervical carcinoma. In this review, epidemiological and molecular data are combined to present a model for HPV-induced cervical carcinogenesis. The impact of current knowledge regarding diagnostic and therapeutic approaches is shown, i.e. the use of HPV tests in cervical cancer screening, in the management of atypical smears of uncertain diagnosis and in smears indicative of mild dysplasias, as well as in follow-up examinations during and after therapy. In addition, the value of the two most frequently used HPV detection systems, polymerase-chain reaction (PCR) and hybrid capture (HC) analysis, is discussed. PMID- 11037342 TI - In situ reverse-transcriptase polymerase chain reaction demonstration of the EWS/FLI-1 fusion transcript in Ewing's sarcomas and peripheral primitive neuroectodermal tumors. AB - It is now widely accepted that the EWS/FLI-1 fusion transcript is associated with tumors of the Ewing family. To test whether it is possible to detect the fusion transcript by means of combining polymerase chain reaction (PCR) methodology and immunohistochemistry, we investigated tumors of the Ewing family using in situ reverse transcriptase (RT)-PCR. We were able to demonstrate the t(11;22) fusion transcript in five of six cases of Ewing's sarcoma and four of four peripheral primitive neuroectodermal tumors. These results were confirmed using fluorescence in situ hybridization in seven tumor samples. In situ RT-PCR-labeled fusion transcripts were found in virtually all tumor cells within a given sample, indicating that each cell possessed the t(11;22) transcript. We conclude from these results that in situ RT-PCR can be used for the rapid detection of EWS/FLI 1 fusion transcripts in biopsy material. The findings also suggest that all cells of the tumors of the Ewing family carry the EWS/FLI-1 fusion transcript. PMID- 11037343 TI - p53 gene status and expression of p53, mdm2, and p21Waf1/Cip1 proteins in colorectal cancer. AB - Abrogation of the normal p53 pathway is the most common molecular alteration in human cancer. p53 Gene status can be potentially assessed through the expression of proteins known to be activated by the wild-type p53 (wt p53) system, such as mdm2 and p21Waf1/Cip1. In this study, the frequency of mdm2, p21Waf1/Cip1, and p53 protein expression was investigated using immunohistochemistry (IHC) in 88 colorectal carcinomas (CRCs). The relationship between these expressions and p53 status was examined. p53 status and the immunophenotypes characterizing these tumors were correlated with standard prognostic variables. Mutation of p53 was detected using single-strand conformational polymorphism (SSCP) analysis and sequencing. Concordance between p53 gene status and p53 immunoreactivity was seen in 62 of 88 (70.45%) carcinomas. Mdm2 expression was found in 22 of 45 (48.88%) and 5 of 43 (11.62%) of the tumors with wt p53 and mutated p53 (P<0.0001), respectively. Predominantly, higher p21Waf1/Cip1 expression was associated with wt p53 (P<0.001). All wt p53 cases that expressed mdm2 also expressed p21Waf1/Cip1. These results suggest that there is a subgroup of CRCs in which p53 is functionally active, inducing transcription of mdm2 and Waf1/Cip1. Their combined evaluation may provide important clues for planning adjuvant systemic therapy and gene therapy based on the restitution of p53 function. However, no significant association was found between the immunophenotypes and the standard prognostic variables investigated. PMID- 11037344 TI - Patterns of chromosomal imbalances in benign solitary fibrous tumours of the pleura. AB - Solitary fibrous tumours (SFTs) of the pleura, in contrast to malignant mesothelioma, occur independently of previous asbestos exposure. They are benign tumours, but may recur if the stalk to the adjacent pleural or lung tissue remains in situ during surgical removal. The molecular pathology of SFTs is largely unknown. We used comparative genomic hybridisation (CGH) to characterise 12 localised SFTs and 12 predominantly sarcomatoid mesotheliomas. Fifty-eight percent of the investigated SFTs did not show any chromosomal imbalances. The most frequent defects were losses on chromosome arms 13q (33%), 4q and 21q (17% each). Significant gains were seen at chromosome 8 and at 15q in two cases each. There was no correlation between tumour size and molecular pathology findings. In contrast, 75% of the mesotheliomas carried chromosomal defects. On average, the mesotheliomas showed over three times as many defects per tumour as the SFTs. Localisation of several frequent losses and gains were similar to those of the SFTs. Therefore, in individual cases, a clear distinction between SFTs and sarcomatoid mesotheliomas is not possible based on CGH analysis alone. Further molecular characterisation of this rare tumour entity will be necessary to elucidate possible genes involved in early tumorigenesis. PMID- 11037345 TI - Regulation of proliferation and apoptosis in sporadic and hereditary medullary thyroid carcinomas and their putative precursor lesions. AB - C-cell hyperplasia (CCH) and medullary thyroid carcinoma (MTC) in patients affected by germline mutations of the RET oncogene represent an exceptional opportunity to study the regulation of proliferation and apoptosis during tumour initiation and progression. In 56 specimens [CCH, n=1; MTC with CCH, n=26; MTC, n=20; lymph-node metastasis (LNM), n=9] from 46 patients [multiple endocrine neoplasia type 2a (MEN2a), n=24; MEN2b, n=2; familiar MTC (FMTC), n=4; sporadic MTC, n-16] and 3 cases of non-neoplastic CCH, proliferation activity (MIB1), the rate of apoptosis [dUTP nick end labelling (TUNEL)] and expression of p53, bcl-2, bcl-x and bax were investigated and compared with clinical data. In MEN associated CCH and small MTC, bcl-2 was strongly expressed, bcl-x was moderately expressed and bax was only weakly expressed. Advanced tumours and LNM did show a more heterogeneous bcl-2 staining accompanied by an increased bax expression and accelerated proliferation. The rate of apoptosis was extremely low in all investigated tumours. P53 was detectable in three patients with rapidly growing and extensively metastasising MTC. No somatic p53 mutations were found. Hereditary MTC with germline RET mutations at codon 918 (MEN2b) and codon 634 revealed a bias towards a higher proliferation activity at a younger age and are more frequently accompanied by LNM. CCH and MTC are characterised with a preponderance of bcl-2 as a factor blocking the programmed cell death. While MTC, in general, is a slowly growing tumour, a minority of tumours do progress rapidly with high proliferation. The factors leading to an accelerated tumour progression do not seem to take their effect via the regulation of apoptosis. Certain alterations of RET are supposed to have a direct or indirect implication on proliferation and, because of this, an effect on the clinical course. PMID- 11037346 TI - Detection of gonadotropin-releasing hormone receptor in normal human pituitary cells and pituitary adenomas using immunohistochemistry. AB - Gonadotropin-releasing hormone (GnRH), which is a well-known regulator of gonadotroph function, has recently been considered to be a paracrine factor involved in the control of somatotroph, lactotroph, and corticotroph cells. GnRH action is initiated by binding to a specific cell surface receptor, the gonadotropin-releasing hormone receptor (GnRHR), which is expressed by follicle stimulating hormone/luteinizing hormone (FSH/LH) cells. Using in situ hybridization techniques, GnRHR messenger ribonucleic acid (mRNA) has recently been detected in normal human anterior pituitary gland and in various pituitary adenomas, including FSH/LH-cell, growth hormone (GH)-cell, adrenocorticotropic hormone (ACTH)-cell, and null-cell adenomas. However, immunohistochemical studies indicating the specific cell distribution of GnRHR in normal pituitary cells have never been reported. The aim of the present investigation was to evaluate the immunohistochemical expression of GnRHR in different types of normal pituitary cells and related tumors. Using double-label immunohistochemical techniques on formalin-fixed and paraffin-embedded tissues and specific antibodies directed against pituitary hormones and GnRHR, we found GnRHR immunoreactivity not only in FSH/LH cells, but also in GH- and thyroid-stimulating hormone (TSH) cells. GnRHR was detected in FSH/LH-cell, GH-cell, mixed GH- and prolactin (PRL)-cell, and alpha-subunit (alpha-SU)/null-cell adenomas. The findings of this study suggest that the interaction between GnRH and GnRHR may play a role in paracrine/autocrine regulation of different types of normal pituitary cells and pituitary adenomas. PMID- 11037347 TI - CD99 immunoreactivity in gastrointestinal and pulmonary neuroendocrine tumours. AB - Although considered a specific marker for Ewing's sarcoma/peripheral neuroectodermal tumour, the MIC2 gene product (CD99) has been immunolocalised in a variety of human tumours. The present study evaluated immunohistochemically the prevalence of CD99 expression in a series of 68 neuroendocrine tumours of different gastrointestinal and pulmonary sites. We now report on membrane and/or granular cytoplasmic immunoreactivity in 25% of these tumours, independent of their anatomical sites. In lung neuroendocrine tumours, CD99 was preferentially confined to typical carcinoids (P=0.009). A statistically significant relationship was observed between the number of CD99 positive cells but not the immunostaining patterns and the presence of local invasion and/or distant metastases (P<0.001). Moreover, there was a tendency for CD99-reactive tumours to show a reduced proliferative activity expressed by a Ki67 index of 2% (P=0.119). The number of CD99 immunoreactive cells or patterns of immunoreactivity did not correlate with the presence of associated clinical syndrome or particular hormonal immunostaining. Although the molecular basis underlying CD99 expression in neuroendocrine tumours is still poorly understood, our data suggest that CD99 may be involved in cell-to-cell adhesion of neuroendocrine tumour cells and in downregulation of their proliferative activity. PMID- 11037348 TI - Patterns of keratin polypeptides in 110 biphasic, monophasic, and poorly differentiated synovial sarcomas. AB - Synovial sarcoma is a mesenchymal neoplasm of unknown histogenesis that shows various degrees of epithelial differentiation. It is known to contain simple epithelial keratins, and the possibility of complex epithelial keratin expression has been suggested. In this study, we immunohistochemically examined 110 well documented synovial sarcomas including 44 biphasic, 48 monophasic, and 18 poorly differentiated (undifferentiated, highly mitotically active) tumors for 11 different keratin (K) polypeptides of the Moll catalogue. The epithelia of biphasic synovial sarcomas showed consistent, extensive reactivity for K7, K8, K14, K18, and K19. Other keratins seen in the epithelia of biphasic tumors included K17 (variable, in 77%), K13 (25%), K16 (23%), and K6 (24%) in the minority of biphasic tumors, predominantly in stratified-appearing epithelia. K10 was detected only focally in one case that showed keratinizing squamous differentiation. Focal expression of K20 was seen in 27% of cases. Monophasic synovial sarcomas had a more limited keratin repertory. Simple epithelial keratin positivity was detected, usually focally for K7 (79%), K19 (60%), K8 (45%), and K18 (46%). Two cases showed more extensive keratin positivity in the spindle cells. The monophasic tumors showed limited positivity for complex epithelial keratins: K14 (28%) and K17 (10%). K20 was detected focally in 6% of the monophasic tumors; other keratins were not detected. The poorly differentiated synovial sarcomas showed limited simple epithelial keratin reactivity, usually limited to scattered cells: K19 (61%), K7 (50%), K18 (47%), K8 (33%), but five cases showed more extensive positivity. Complex epithelial keratins were scant: K14 in one case and K17 in two cases. The immunoreactivity of capillary endothelia seen for K7 and K18 (but not for K8 and K19 with the antibodies used) is a potential diagnostic pitfall, and may cause overdiagnosis of synovial sarcoma if not properly recognized. In summary, we show complex patterns of keratins in synovial sarcoma, especially in the biphasic tumors. Such patterns establish a baseline in differential diagnostic considerations, and give an insight into the complex epithelial differentiation of this enigmatic mesenchymal tumor. PMID- 11037349 TI - Benign metastasizing leiomyoma of the uterus: documentation of clinical, immunohistochemical and lectin-histochemical data of ten cases. AB - The clinical histories of 10 women suffering from benign metastasizing leiomyoma (BML) after hysterectomy and information on lung lesions detected in these women are presented, together with corresponding data for 2 women with metastasizing leiomyosarcoma of the uterus for comparison: gross appearance, survival, and light microscopical, immunohistochemical and lectin-histochemical findings are reported. All patients with BML had undergone hysterectomy for uterus leiomyomatosus without any detection of sarcomatous lesions in the uterus wall. After a median period of 14.9 years intrapulmonary masses were detected by imaging techniques. On average, six nodules with a mean diameter of 1.8 cm were seen. Resection of the lesions was performed in all cases. The immunohistochemical and lectin-histochemical examination of the tumors included analysis of the proliferation-associated protein Ki-67, the p53 protein, estrogen and progesterone receptor, sarcolectin as an indicator of the presence of lymphokine macrophage migration inhibitory factor, antibodies and the labeled protein to assess galectin (galactoside-binding animal lectin)-dependent parameters, analysis of tumor vascularization (CD-34), and expression of bcl-2, vimentin, smooth muscle actin, desmin, and keratin. The lesions were characterized by low proliferation activity of 2.9% (measured with Ki-67), frequent hormone receptor expression (8 of the 10 cases presented hormone specific receptors), low to moderate vascularization compared with metastases from the two uterine sarcomas, remarkable p53 overexpression and frequent expression of the lymphokine, the galectins and accessible binding sites. The median survival of the BML patients was 94 months after excision of the intrapulmonary lesions, and the maximum survival of the two sarcoma patients was 22 months. The results recorded in this patient sample with the methodology applied suggest that benign metastasizing leiomyomas are a slow-growing variant of leiomyosarcoma of the uterus, which becomes clinically apparent at a young age and progresses with low velocity. PMID- 11037350 TI - Significant association of strictures and internal fistula formation in Crohn's disease. AB - Intestinal inflammation in Crohn's disease (CD) may be complicated by the occurrence of strictures and fistulae. The pathogenesis of fistula formation is unknown. We therefore wanted to determine whether mechanical factors might contribute to the development of fistulae. Furthermore, we tried to define the path of internal fistulae through the muscular layer. For this purpose, surgical resection specimens from 42 consecutive patients with CD were prospectively studied. In gross examination the whole bowel was cut into circumferential cross sections 0.3 cm thick. Abnormal areas were histologically examined. Strictures were found in 38 patients (90.5%), and fistulae were observed in 27 (64.3%) patients. In 11 (40.7%) specimens fistulae were found within a stricture, in 15 (55.6%) at the proximal end, and in 1 (3.7%) no stricture was found. In 7 (25.9%) cases with fistulae, herniated mucosa was found within the muscularis propria or the subserosa. In 7 (25.9%) cases a blood vessel was identified near a fistula traversing the muscularis propria. From these findings we conclude that that mechanical factors may contribute to fistula formation. This is further supported by the fact that fistulae appear to traverse the muscular layer along piercing vessels. PMID- 11037351 TI - AgNOR quantity as a prognostic tool in hyperplastic and neoplastic parathyroid glands. AB - Prediction of evolution of secondary hyperplasia and tumours of the parathyroid glands is still a problem in histopathology. To assess whether the quantity of silver-stained nucleolar organiser region (AgNOR) proteins might be used as a prognostic tool in parathyroid pathology, a standardised AgNOR analysis has been performed on 19 cases of parathyroid hyperplasia caused by secondary hyperparathyroidism (PH), 8 cases of adenoma (PA) and 10 cases of carcinoma (PC). Clinico-pathological data and follow-up information were available. On formalin fixed and paraffin-embedded sections, the visualisation and quantification of AgNORs were achieved according to the 1995 guidelines of the Committee on AgNOR Quantification. Then, the mean area (square micrometres) of AgNORs per nucleus (NORA) was evaluated by means of an image analyser and specific softwares. After testing the normal distribution of NORA values, statistical parametric tests were utilised; Kaplan-Meier and Cox multivariate analyses were also performed. In parathyroid lesions, a progressive increase of mean NORA values was observed from PH (2.895 microm2; SE 0.171) through PA (3.638 microm2; SE 0.125) to PC (4.701 microm2; SE 0.179); these differences were highly significant (P<0.001), although some degree of overlap was found among single NORA values. A significantly higher mean NORA value was revealed in PC with distant metastases than was noted in cases with no current clinical evidence of disease progression. Furthermore, a significantly (P<0.001) higher mean NORA value was encountered in the group of PH with recurrences (3.600 microm2; SE 0.106) than in nonrecurrent PH (2.261 microm2; SE 0.087). Multivariate analyses indicated that the NORA value was an independent prognostic parameter determining the risk of recurrence in PH. We suggest that AgNOR quantity may be a promising additional tool for predicting the biological behaviour of parathyroid lesions. PMID- 11037352 TI - Interstitial pneumonia in Hermansky-Pudlak syndrome: significance of florid foamy swelling/degeneration (giant lamellar body degeneration) of type-2 pneumocytes. AB - Although usual interstitial pneumonia (UIP)-like IP has been known as the most serious complication of Hermansky-Pudlak syndrome (HPS), its pathologic features and pathogenesis are poorly understood. We investigated biopsied and autopsied lung tissues from five patients who died of UIP-like IP associated with HPS (HPSIP). The salient histopathologic features of HPSIP observed were: (1) alveolar septa displaying florid proliferation of type-2 pneumocytes (2PCs) with characteristic foamy swelling/degeneration; (2) patchy fibrosis with lymphocytic and histiocytic infiltration centered around respiratory bronchioles, occasionally showing constrictive bronchiolitis; and (3) honeycomb change without predilection for the lower lobes or subpleural area. Those peculiar 2PCs were histochemically characterized by the over accumulation of phospholipid, immunohistochemically by a weak positivity for surfactant protein, and ultrastructurally by the presence of numerous giant lamellar bodies that compressed the nucleus with occasional cytoplasmic disruption, together suggesting a form of cellular degeneration with an over accumulation of surfactant (giant lamellar body degeneration). The present study strongly indicates that there is a basic defect in the formation/secretion process of surfactant by the 2PCs in HPS, which may well be the triggering factor for the HPSIP development. Other factors, such as macrophage dysfunction, may be working synergistically for further acceleration of the inflammatory process. PMID- 11037353 TI - Induction of adenocarcinoma containing myoepithelial cells in rat submandibular gland by 7,12-dimethylbenz(a)anthracene. AB - In an attempt to induce adenocarcinoma containing myoepithelial cells (MECs) in the rat submandibular gland, we injected 7,12-dimethylbenz(a)anthracene (DMBA) dissolved in acetone into the glands of rat pups at the age of 10 days. In both male and female pups, the glands, including their developing terminal secretory units, contained far greater numbers of cells positive for proliferating cell nuclear antigen (PCNA) than did adult glands. A single administration of 1% DMBA (0.05 ml/130 g b.w.) did not produce adenocarcinoma, but did induce occasional sarcomas, such as rhabdomyosarcoma and fibrosarcoma, in 2 months. Most glands regenerated with minimal scar formation. Microscopically, these glands were atypical in that they contained increased numbers of PCNA-positive cells, underdeveloped granular ducts, and striated ducts surrounded by MECs positive for alpha smooth muscle actin (alphaSMA). Though these features were also observed in the regenerated glands after acetone injection, the number of PCNA-positive cells was relatively high in the glands of DMBA-treated females, especially in the terminal secretory unit. The second DMBA injection at 10 weeks of age produced adenocarcinoma made up of alphaSMA-positive MECs and keratin 19-positive duct cells. Such MEC-associated adenocarcinoma was induced in the glands of more than half the female but not the male animals. Replacement of either of the double DMBA treatments with acetone, or DMBA treatment, single or double, of adult glands did not produce adenocarcinoma, but did produce sarcoma and squamous cell carcinoma. These results suggest that (1) at least two genetic mutations are necessary for induction of adenocarcinoma with MECs in the rat submandibular gland, (2) the mutation is efficiently introduced to pup glands whose terminal secretory units exhibit extreme proliferative activity, and (3) the second mutation is difficult to introduce in male glands, whose proliferative activity is relatively low, and/or transformed cells need some female hormone after the mutation to propagate. PMID- 11037354 TI - Different frequency of cilia with transposition in human nasal and bronchial mucosa. A case of acquired ciliary dyskinesia. AB - Nasal and bronchial cilia and spermatozoa of a patient with a high clinical suspicion of a ciliary dyskinesia syndrome were ultrastructurally studied and quantified. Defective cilia showed two types of axonemal patterns: 9d+0s and 8d+1d. Of these, 9d+0s cilia prevailed in the proximal region, whereas 8d+1d prevailed in the distal region. Translocation of a peripheral doublet to the central position occurred at the middle region of cilia lacking the central pair, probably to compensate for its absence. Quantitative analysis showed that the percentages of anomalous cilia were 5.32+/-0.93 in nasal samples and 43.17+/-2.34 in bronchial samples. Spermatozoa without the central pair or with a translocated microtubular doublet were rarely observed, but a variety of nonspecific defects were seen. Even though transposition is generally considered to be an inherited ciliary defect and one of the causes of primary ciliary dyskinesia, in this case quantitative ultrastructural analysis and clinical data indicate that this is an acquired ciliary defect. PMID- 11037355 TI - So-called mesothelial/monocytic incidental cardiac excrescences obtained during valve replacement surgery: report of three cases and literature review. AB - We present three cases of so-called mesothelial/monocytic incidental cardiac excrescences (MICE) of the heart and a brief review of related literature. Case 1 was a 51-year-old woman who underwent mitral- and aortic-valve replacement. A tissue sample was submitted as a thrombus attached to the left atrial endocardium. Case 2 was a 69-year-old woman who underwent mitral-valve replacement. The sample was incidentally obtained as whitish clot-like fragments, but its exact origin was not known. Case 3 was a 68-year-old woman who underwent mitral-valve replacement for suspected infective endocarditis. The sample adherent to the pericardium was removed after valvular surgery. Histologically, these lesions were composed of a mixture of plump histiocytoid cells, a papillary arrangement of cuboidal cells, various sized vacuoles, and fibrin. The nests of cuboidal cells resembled cancer cells but showed features of mesothelial cells and no proliferative activity, immunohistochemically or ultrastructurally. In all cases, a suction tube placed in the left atrium was occasionally used to remove overflowing intrapericardial fluid during the surgery. The tip of the suction tube was covered with spiral wire, which is likely to transfer the stripped pericardial mesothelial cells to the left atrium. The significance of MICE is their possibility of being misdiagnosed as metastatic carcinoma by pathologists and a risk of arterial embolization by mesothelial debris clinically. PMID- 11037356 TI - Health and society in the 21st century. PMID- 11037358 TI - Liveliness. AB - OBJECTIVE: The objective of this study is to explore the significance of the experience of liveliness in psychotherapeutic interactions and its relevance to the practice of psychotherapy. METHOD: Stern's notion of 'vitality affects' and Emde's concept of a 'primary affective core' are employed in developing a concept of liveliness, or an enlivening-deadening axis of experience. A critique of Freud's 'principles of mental functioning' is made in the light of this concept. Clinical examples are provided as illustrations of the relevance of considering the 'sense of liveliness', and its sustainability, in psychotherapy. RESULTS AND CONCLUSIONS: A sense of liveliness relates closely to activity within a system of interpersonal resonance with non-linear characteristics. The experience of, and responses to, vitality affects may be an important basis of a sense of liveliness. Sudden shifts towards experiences of deadness are a matter for concern in psychotherapy. This sphere of experience, although occurring largely outside verbal awareness, may constitute a distinct type of mental process. Three types of mental activity or process are postulated: (i) emergent, pre representational activity characterised by the sense of liveliness; (ii) Play related thought or activity typically experienced as enlivening; and (iii) work related or adaptational thought or activity. PMID- 11037357 TI - Attention deficit hyperactivity disorder: current progress and controversies. AB - OBJECTIVE: Attention deficit hyperactivity disorder (ADHD) is a common chronic and disabling condition in children. This paper reviews the taxonomic issues and the major comorbid conditions, neurobiological correlates, treatment and public health issues associated with ADHD. METHOD: Pertinent recent papers are reviewed from the psychological and psychiatric literature. RESULTS: The two major taxonomies now define a similar group of children with ADHD of a combined type/hyperkinetic disorder. Advances in the understanding and treatment of ADHD demonstrate the complex multidimensional links between neurobiology, psychology and behaviour. Careful assessment of individual factors in treatment planning and ongoing monitoring of psychostimulant medication treatment in the longer term are recommended. CONCLUSIONS: There is much still to learn about ADHD, and increased levels of clinical research and treatment resources are required. PMID- 11037359 TI - Adolescent depressive disorder: a population-based study of ICD-10 symptoms. AB - OBJECTIVE: Earlier studies have suggested that symptoms of depressive disorder in adolescents may differ from those found in adults. Even so, diagnostic criteria developed in adults have come to be widely applied to younger subjects. This study examines the frequency of ICD-10 symptoms in depressive disorder and their association with severity in a large community sample of adolescents aged 15 to 18 years. METHOD: A six-wave prospective study of adolescent health and emotional wellbeing in 2032 Australian secondary school students provided an opportunity to conduct a two-phase study of adolescent onset depression. A self-administered computerised form of the revised Clinical Interview Schedule (CIS-R) was used as a first phase diagnostic measure. Second phase assessment using the Composite International Diagnostic Interview (CIDI) allowed the delineation of a group fulfilling criteria on both instruments. The ICD-10 symptoms and severity profiles for depression were generated with standard algorithms. RESULTS: 1947 (95.8%) out of 2032 subjects in the designated sample completed phase 1 assessment at least once. Participation rates at phase 2 interviews were 93%. Over the 30-month study period 69 subjects (10 male, 59 female) fulfilled criteria for ICD-10 depressive episodes on both the CIS-R and CIDI. Thirty-one per cent (n = 21) had experienced a severe episode, 46% (n = 32) moderate and 23% (n = 16) mild episodes. Loss of interest and pleasure, decreased energy and fatigue, sleep disturbance, suicidal ideation and diminished concentration most clearly distinguished adolescents with depressive disorder from controls. Self reproach and guilt, psychomotor agitation and/or retardation and appetite disturbance with weight change showed the clearest increase in frequency with increasing severity of episode. The somatic syndrome was reported by close to one in three of those with a severe depressive episode, but was uncommon in those with mild and moderate episodes. CONCLUSIONS: The ICD-10 diagnostic criteria are applicable to depressive disorder in older adolescents. With the exception of depressed mood, found in one in five non-cases, all other symptoms were common in cases and uncommon in non-cases. Practitioner awareness of symptoms indicating the presence and severity of disorder should enhance early identification and choice of treatment in adolescent depression. PMID- 11037360 TI - The impact of the organisation of mental health services on the quality of assessment provided to older patients with depression. AB - OBJECTIVE: The objective of this study was to examine the impact of the organisation of mental health services on the quality of medical and psychiatric assessment provided to patients with depression over 50 years of age. METHOD: A retrospective clinical audit of 99 patients with primary depressive disorders who were over 50 years of age was used. These patients were assessed initially by specialised psychogeriatric outpatient and community services (44%), community based adult mental health services (35%) or an inpatient service (21%). At 2-3 years follow up, clinical outcomes were rated by treating physicians and included current depression status, cognitive and medical status, course of illness since initial assessment and current living circumstances. RESULTS: Patients who were assessed by the community-based adult mental health service received the least comprehensive assessment. Although these patients were more likely to be living independently, they tended to have the poorest depression outcome. Patients who were assessed by the specialised or inpatient services received more comprehensive initial assessment and better coordinated long-term care. Although these patients had more medical and cognitive comorbidity they had better overall depression outcomes. CONCLUSIONS: Within a service system that determines access according to an arbitrary age of onset, patients with depression receive the best assessment from specialised psychogeriatric services. However, patients with an early age of onset, more chronic disorders and poor outcomes are treated largely within community-based adult services. Psychiatric services need to ensure that all older patients with depression receive appropriate biomedical and psychosocial assessment, as well as continuity of medical and psychological treatment. PMID- 11037361 TI - Effect of somatic comorbidity on alleviation of depressive symptoms. AB - OBJECTIVE: The aim of this study was to investigate whether somatic comorbidity (SC) impedes recovery from depression. METHOD: The study design was naturalistic. Diagnosis of depression was confirmed by means of the Structured Clinical Interview for DSM-III-R (SCID). Changes in the symptom scales for those patients with somatic comorbidity (n = 75) were compared with corresponding changes in depressive patients without somatic comorbidity (n = 41) in a 6-month follow up. RESULTS: Measured on the Hamilton and Beck scales, recovery rates of those with SC was only slightly lower to that of the others. The difference was statistically significant only in relation to the Hamilton scale. Forty-four per cent of those with SC and 42% of the other patients recovered from their depression (BDI score < 10 on follow up). Logistic regression analysis showed no independent association between recovery and somatic comorbidity. CONCLUSIONS: Moderate somatic comorbidity has only a minor effect on recovery from depression. PMID- 11037362 TI - Chronic fatigue syndrome and Australian psychiatry: lessons from the UK experience. AB - OBJECTIVE: The aim of this paper is to outline the opportunities and dangers the chronic fatigue syndrome (CFS) issue presents to Australian psychiatry. METHOD: The scientific literature of the last 50 years on CFS in adults was reviewed and samples of recent media portrayals of CFS in the UK and Australia were collected. The author has worked in both the UK and Australia managing adult CFS patients in specialist outpatient consultation-liaison (C-L) psychiatry settings. RESULTS: Chronic fatigue syndrome has been at the heart of an acrimonious debate in the UK, both within the medical profession and in the wider community. UK psychiatry has been drawn into the debate, at times being the target of strong and potentially damaging criticism, yet UK psychiatry, especially the C-L subspecialty, has played a crucial role in clarifying appropriate research questions and in devising management strategies. The issue has served to enhance and broaden psychiatry's perceived research and clinical role at the important medicine-psychiatry interface in that country. CONCLUSIONS: Handled properly, the CFS issue offers Australian psychiatry, especially C-L psychiatry, an opportunity to make a useful contribution to patient care in a clinically difficult and contentious area, while at the same time serving to help broaden psychiatry's scope in the Australian medical landscape. PMID- 11037363 TI - Issues associated with the identification of cognitive change following coronary artery bypass grafting. AB - OBJECTIVE: Coronary artery bypass grafting (CABG) is a surgical procedure used to treat individuals with ischaemic heart disease and to relieve angina. Disruption to the central nervous system (CNS) has frequently been reported by patients who have undergone CABG. METHOD: The following paper is a review of the literature that has examined the effects of CABG on the CNS. RESULTS AND CONCLUSIONS: It becomes apparent that issues about the incidence and severity of post-CABG cognitive decline are still unresolved. First, the cause of post-CABG CNS change has not yet been established, although the presence of changes to brain microvasculature as a result of the presence of microemboli appears to be a likely factor. Second, while some studies have reported high rates of poor performance on neuropsychological tests postoperatively, these reports are often subject to confounds such as variability in postoperative testing intervals, the definition of decline and the neuropsychological test batteries used. Finally, improvements in surgical techniques and changes in patient characteristics may have changed the real nature and prevalence of post-CABG cognitive decline. The review finishes with a series of recommendations for the neuropsychological study of CABG. PMID- 11037364 TI - Prevalence of psychological distress and use of support services by cancer patients at Sydney hospitals. AB - OBJECTIVE: The study aimed to estimate the prevalence of anxiety and depression within a cross section of cancer patients in the Sydney region, and to assess the use of and degree of satisfaction with available support services. METHOD: A survey was conducted at oncology outpatient departments of four Sydney hospitals. Participants completed the Hospital Anxiety and Depression Scale (HADS) and a questionnaire seeking information on their use of patient support services, as well as demographic and clinical information. RESULTS: A total of 504 valid questionnaires were returned. Using a cut-off score of 11 on the HADS, the prevalence of clinically significant anxiety and depression was 11.5% and 7.1% respectively; 17% of patients had received individual counselling while 6.5% had attended support groups. The majority of patients who had attended counselling or support groups reported them to have been 'extremely' or 'reasonably' helpful (86% and 83% respectively). Of the patients who were experiencing clinically significant anxiety or depression, 75% had not received any counselling or psychological treatment. The main factors which predicted clinically significant anxiety or depression were: restricted activity levels, advanced disease, a non English-speaking background and being female. CONCLUSIONS: While the prevalence of clinically significant anxiety or depression detected by the HADS was reasonably low, a substantial number of possible cases were identified. The majority of affected patients were not accessing counselling or psychological treatment. Systematic screening of oncology patients at hospital entry might enable more immediate identification of clinically affected patients, who could then be referred for further testing or psychological treatment. PMID- 11037365 TI - Characterising psychosis in the Australian National Survey of Mental Health and Wellbeing Study on Low Prevalence (psychotic) Disorders. AB - OBJECTIVE: This study examines the factorial structure of symptoms and signs in psychosis in data from the Study on Low Prevalence (psychotic) Disorders which is part of the National Survey of Mental Health and Wellbeing, Australia 1997-1998. METHOD: The present study examined a wide variety of symptoms taken from the Schedules for Clinical Assessment in Neuropsychiatry items and the substance use items in the Diagnostic Interview for Psychosis, an instrument specially constructed for the national study. The instrument was applied to 980 community and hospital subjects with a wide range of psychotic illness diagnoses. The data were factor analysed and scales of 'domains of psychopathology' derived. RESULTS: The data were best fitted by five principal factors ('domains') which can be approximately labelled dysphoria, positive symptoms, substance use, mania and negative symptoms/incoherence. These factors together explained 55.4% of variance in symptoms. Solutions with more numerous factors did not improve the representation. CONCLUSION: The five domains successfully characterise a large part of the variance in psychopathology found in the present study of low prevalence (psychotic) disorders. The approach allows sufferer's symptom range and severity to be well expressed without multiple comorbid diagnoses or the limits imposed by categorical diagnosis. Knowledge of alternative dimensional representations of psychopathology may usefully complement our use of categories, enhance awareness of symptoms and ensure that important psychopathology is heeded in practice and research. PMID- 11037367 TI - A cognitive-behavioural, group-based intervention for social anxiety in schizophrenia. AB - OBJECTIVE: The objective of this study was to investigate the efficacy of group based cognitive-behavioural therapy (CBT) for social anxiety in schizophrenia. METHOD: Patients with schizophrenia (20) with comorbid social anxiety were randomly assigned to the group-based CBT or wait-list control condition. Pre-, post- and 6-week follow-up ratings included measures of social anxiety and avoidance, mood and quality of life. RESULTS: The intervention group improved on all outcome measures and the control group showed no change in symptomatology. CONCLUSIONS: Group-based CBT is effective in treating social anxiety in schizophrenia. PMID- 11037366 TI - Community treatment orders: relationship to clinical care, medication compliance, behavioural disturbance and readmission. AB - OBJECTIVE: The objective of this study was to investigate the readmission rate, and the level of patient disturbance and community care associated with readmission following Community Treatment Orders (CTOs) in New South Wales, Australia. METHOD: The readmission rates of all patients given CTOs within a 4 year period and a matched comparison group were investigated. The following factors were compared before, during and following a CTO: medication non compliance, number of clinical services and duration of disturbed behaviour preceding hospitalisations. RESULTS: Of 123 patients on CTOs (mean length, 288 days; SD, 210 days), 38 were readmitted during the CTO, the majority in the first 3 months and a further 21 patients were readmitted following termination of the CTO. Evidence of lower severity of illness in the comparison patients prevented meaningful evaluation of the readmission rates of the two groups. While on CTOs, patients receiving depot medications showed high compliance and a significantly reduced readmission rate compared with that of patients receiving oral medications. In the 2 months prior to hospitalisations during CTOs, compared with those before or after CTOs, patients received more frequent consultations and showed a shorter duration of medication non-compliance and disturbed behaviour. The level of services in the 3 months following discharge were comparable for patients on CTOs and the comparison group. CONCLUSIONS: CTOs may reduce rehospitalisations by use of depot medication. Earlier and possibly more frequent readmissions in the CTO group shortened the disturbance associated with illness recurrence. It would appear that to establish a control group with equivalent severity of disorder necessary to evaluate the impact of CTOs requires a random allocation design. PMID- 11037368 TI - Enhancing case managers' skills in the assessment and management of antipsychotic medication side-effects. AB - OBJECTIVE: Our goal was to reduce the prevalence of antipsychotic medication side effects by providing a short-term training program on the assessment and management of side-effects to case managers. METHOD: Forty-four patients in receipt of community-based mental health services were allocated to comparison (n = 20) and intervention (n = 24) groups based on the health service district in which they resided. While case managers working with the intervention group attended a short-term training program to improve their assessment and management of neuroleptic side-effects, case managers providing services to the comparison group received no additional training. Side-effects were assessed pre- and postintervention using the Liverpool University Neuroleptic Side-effect Rating Scale (LUNSERS). RESULTS: A reduction in the overall prevalence of side-effects in both groups was observed, however, only those patients in the intervention group reported a statistically significant reduction in mean side-effect scores between the pre- and postmeasures (Wilcoxon Matched Pairs Signed-ranks Test, z = 2.8411, two-tailed, p < 0.01). In addition, qualitative data collected during the second survey revealed that patients in the intervention group had acquired some positive management strategies for dealing with unwanted side-effects. The strategies were elicited from eight different patients distributed across six of the 12 case managers who took part in the training program. CONCLUSIONS: Training cases managers in the assessment and management of side-effects may help to reduce their impact on the lives of people prescribed neuroleptic medication. PMID- 11037369 TI - Youth suicide trends in Australian metropolitan and non-metropolitan areas, 1988 1997. AB - OBJECTIVE: The objective of this study was to examine trends in suicide among 15 34-year-olds living in Australian metropolitan and non-metropolitan areas between 1988 and 1997. METHOD: Suicide and population data were obtained from the Australian Bureau of Statistics. We calculated overall and method-specific suicide rates for 15-24 and 25-34-year-old males and females separately, according to area of residence defined as non-metropolitan (< or = 20,000 people) or metropolitan. RESULTS: Between 1988 and 1997 suicide rates in 15-24-year-old non-metropolitan males were consistently 50% higher than metropolitan 15-24-year olds. In 1995-1997, for example, the rates were: 38.2 versus 25.1 per 100,000 respectively (p < 0.0001). The reverse pattern was seen in 25-34-year-old females with higher rates in metropolitan areas (7.5 per 100,000) compared with non metropolitan areas (6.1 per 100,000, p = 0.21) in 1995-1997. There were no significant differences according to area of residence in 25-34-year-old males or 15-24-year-old females. Over the years studied we found no clear evidence that suicide rates increased to a greater extent in rural than urban areas. Rates of hanging suicide have approximately doubled in both sexes and age groups in both settings over this time. Despite an approximate halving in firearm suicide, rates remain 3-fold higher among nonmetropolitan residents. CONCLUSION: Non metropolitan males aged 15-24 years have disproportionately higher rates of suicide than their metropolitan counterparts. Reasons for this require further investigation. Hanging is now the most favoured method of non-metropolitan suicide replacing firearms from 10 years ago. Although legislation may reduce method-specific suicide the potential for method-substitution means that overall rates may not fall. More comprehensive interventions are therefore required. PMID- 11037370 TI - The cost of being perfect: perfectionism and suicide ideation in university students. AB - OBJECTIVE: The aim of the study was to assess the relationship between dimensions of perfectionism and suicide ideation in a tertiary student population in Australia. METHOD: The methodology involved 405 students completing the General Health Questionnaire (GHQ-28) which includes a subset of questions which can be used to assess suicide ideation, and the Multidimensional Perfectionism Scale. RESULTS: The presence of suicide ideation was associated with higher scores on total perfectionism and two perfectionism dimensions, and total GHQ scores. There were significant differences between participants with high levels of perfectionism and participants with moderate to low levels of perfectionism on a measure of suicide ideation. Neither gender nor age were associated with differences in the scores, with results indicating high levels of perfectionism may indicate a vulnerability to suicide ideation. CONCLUSIONS: Perfectionism is a valued attribute in high-achieving populations. The question needs to be asked, however, at what cost? The findings indicate that high levels of perfectionism may be associated with an increased vulnerability to suicide ideation. Future research is needed to gain a better understanding of the complex interrelationship between personality and temperament, environmental factors and self-destructive behaviour. PMID- 11037371 TI - Death by hanging: implications for prevention of an important method of youth suicide. AB - AIM: The aim of this study was to identify factors associated with deaths by hanging among young people in Queensland, Australia. METHOD: An examination of coroner's reports for all deaths by hanging of people under 25 years of age that occurred in Queensland in the years 1995 and 1996. RESULTS: All cases were recorded as suicides. Most were males and a quarter were indigenous persons. Half the deaths occurred in regional or rural areas. Unemployment, the experience of personal loss, psychiatric illness and alcohol use were possible precipitating agents. Early warning signs were the onset of uncharacteristic behaviours and threats of suicide. CONCLUSIONS: The private nature of hanging means that there are rarely opportunities to prevent it in the period immediately before the fatal event. Earlier interventions will have to be considered. To prevent hanging as a means of suicide, we need to understand more about the difficulties experienced by some young men who are living in rural areas. We need more information about the cultural problems experienced by indigenous youths in their teenage years. Young people in the justice system may need personal support. There is a pressing need to determine if young people, especially in rural areas, have adequate access to the professional expertise needed to diagnose and treat mental disorders. PMID- 11037372 TI - The perceived utility of six selected measures of consumer outcomes proposed for routine use in Australian mental health services. AB - OBJECTIVE: This paper aims to assess the utility of six measures of consumer outcomes: the Behaviour and Symptom Identification Scale, the Mental Health Inventory (MHI), the Medical Outcomes Study 36-Item Short-Form Survey, the Health of the Nation Outcome Scales, the Life Skills Profile (LSP) and the Role Functioning Scale previously recommended for the routine assessment in Australian mental health services. METHOD: Consumers and service providers were invited through focus group discussions and surveys to describe the perceived utility of these selected measures. RESULTS: All six measures were rated favourably. The qualitative and quantitative findings suggest that the MHI elicited the most positive results of the consumer measures. No observer-rated scale was clearly preferred. CONCLUSION: The qualitative feedback obtained indicated that process and context issues may be as important to the successful use of routine instruments for the measurement of consumer outcomes in clinical practice as the choice of instrument. PMID- 11037373 TI - Measuring disability, need and outcome in Australian community mental health services. AB - OBJECTIVE: This study trialled routine measurement of disability, need and outcome in mental health services within Sydney. METHOD: Fifteen community mental health clinicians with a combined caseload of 283 patients participated in the study. The Health of the Nation Outcome Scales (HoNOS) was used to assess disability and outcome and the patient and staff versions of the Camberwell Assessment of Need (CAN) were used to assess need. RESULTS: The HoNOS and CAN appear to be promising contenders for routine use. Patients receiving assertive case management were rated as having higher levels of disability and need than patients receiving standard case management. Significant change in outcome was demonstrated with the HoNOS. CONCLUSIONS: To ensure the continued measurement of consumer outcome, issues such as staff education, training, and the development of computerised information systems should be addressed. PMID- 11037374 TI - Interrater reliability of the Camberwell Assessment of Need Short Appraisal Schedule. AB - OBJECTIVE: Australian mental health policy aims to introduce evidence-based practice within a community care approach. This aim requires reliable measures that can be used by a wide variety of professionals. The interrater reliability of the Camberwell Assessment of Need Short Appraisal Schedule (CANSAS) was assessed under routine conditions. METHOD: Three interviewer-observer dyads assessed the needs of 14 inpatients and 18 day patients of a psychiatric rehabilitation unit in New South Wales, Australia. RESULTS: Agreement on the identification of an area of need was high. However, agreement was higher on patient ratings than on staff ratings. Correlations on staff ratings of met needs were also moderate (r = 0.53), suggesting discrepancies in rating the level of need. CONCLUSION: Differences in staff ratings may be attributed to ambiguity in the definition of need and levels of need and/or the sources of information used by the rater making the assessment. An approach to establishing an operational definition of need is suggested, and an increase in the number of levels of need is recommended. Implications for Australian mental health policy are noted. PMID- 11037375 TI - Transsexualism in female monozygotic twins: a case report. AB - OBJECTIVE: A case report is presented of a gender identity disorder involving a pair of female monozygotic twins who requested sex reassignment. As far as we know, this case is the first in psychiatric literature and supports a genetic aetiology of this disorder. CLINICAL PICTURE: The patients were two 18-year-old female monozygotic twins who had showed symptoms of transsexualism since early childhood. They had no other physical or psychiatric disorder except borderline mental functioning. TREATMENT: The patients were referred for sex reassignment. OUTCOME: They were lost to follow up after initial evaluation. CONCLUSION: In addition to other possible (yet not confirmed) causes of transsexualism, a genetic basis for this disorder is suggested by this case. PMID- 11037376 TI - Capgras syndrome: possibly more common among the Maori of New Zealand. AB - OBJECTIVE: The report describes an apparently greater incidence of Capgras syndrome among the Maori population compared with the European population, in the most easterly catchment area served by Tauranga Hospital in New Zealand's Bay of Plenty. METHOD: Over the last year we have become aware of five cases of Capgras syndrome in our catchment area. This area (population nearly 21,000) consists of a rapidly expanding new suburb of the city of Tauranga and a rural area extending 55 km east of the city. These figures were compared with those of the westerly catchment area served by Tauranga Hospital, where the psychiatric team is not aware of any examples of Capgras syndrome among their population. The 1996 census figures were obtained in order to calculate a population ethnicity breakdown. RESULTS: Five cases of Capgras syndrome were identified in the most easterly catchment area where 19% of the population identified as Maori, 75% as European and 6% as other or non-specified. All of the cases occurred in Maori patients. This compares with no identified cases of Capgras syndrome in the most westerly catchment area where 12% of the population identified as Maori, 87% as European and 1% as other or non-specified. Four out of five cases were female. Two cases had a history of cannabis use. Three cases had exhibited dangerous behaviour towards family members. CONCLUSIONS: There is an apparently greater incidence of Capgras syndrome among the New Zealand Maori population compared with the European population in the most easterly catchment area served by Tauranga Hospital. In our population Capgras syndrome is a common, not rare, feature of psychotic illness, and the cases support a previously reported association of this syndrome with dangerousness. PMID- 11037377 TI - Response to 'The flight of the wild goose: the psychiatrist as a leader'. PMID- 11037378 TI - Borderline personality disorder. PMID- 11037379 TI - Response to 'Does reducing safety behaviours improve treatment response in patients with social phobia?'. PMID- 11037380 TI - Rethinking Karl Ludwig Kahlbaum's contribution to biological psychiatry. PMID- 11037381 TI - Response to 'Psychostimulants, adult attention deficit hyperactivity disorder and morbid jealousy'. PMID- 11037382 TI - Shine: further notes of discord. PMID- 11037383 TI - Management of acute psychosis. PMID- 11037384 TI - Catatonic and non-catatonic neuroleptic malignant syndrome. PMID- 11037385 TI - Cyproterone for hypersexuality in a psychotic patient with Wilson's disease. PMID- 11037386 TI - Risperidone and tardive dyskinesia: a case of blepharospasm. PMID- 11037387 TI - Myocarditis associated with clozapine treatment. PMID- 11037388 TI - Dimensions and categories: the "big five" factors and the DSM personality disorders. AB - The five-factor model of personality, which has been widely studied in personality psychology, has been hypothesized to have specific relevance for DSM defined personality disorders. To evaluate hypothesized relationships of the five factor model of personality to personality disorders, 144 patients with personality disorders (diagnosed via a structured interview) completed an inventory to assess the five-factor model. Results indicated that the majority of the personality disorders can be differentiated in theoretically predictable ways using the five-factor model of personality. However, while the personality disorders as a whole appear to be differentiable from normal personality functioning on the five factors, the patterns are quite similar across the disorders, a finding that may provide some insight into the general nature of personality pathology but may also suggest problems with discriminant validity. Third, it does not appear that considering disorders as special combinations of features (as might be expected in some categorical models) is more informative than considering them as the sum of certain features (as might be expected in a dimensional model). PMID- 11037389 TI - Qualitative block design analysis in posttraumatic stress disorder. AB - Qualitative and quantitative Block Design performance was examined in Vietnam combat veterans with PTSD diagnoses (n = 23) and Vietnam combat veterans without PTSD or other mental disorders diagnoses (n = 19). Results indicated that PTSD diagnosed veterans committed more single block rotations than the comparison sample, and that their errors occurred more frequently in right hemispace than errors made by the comparison sample. The two groups did not differ in the number of configural errors made, errors committed in left hemispace, or in quantitative performance measures. Findings are suggestive of relative left hemisphere hypoactivation and are congruent with prior research documenting cerebral asymmetries in emotional disorders. PMID- 11037390 TI - Psychological test usage with adolescent clients: survey update. AB - In 1991, Archer, Maruish, Imhof, and Piotrowski presented survey findings based on the responses of a national sample of psychologists who performed psychological assessment with adolescent clients. The current survey was designed to update their results by examining the test use practices reported by 346 psychologists who work with adolescents in a variety of clinical and academic settings. These respondents represented an adjusted survey return rate of 36% and predominantly consisted of doctoral prepared psychologists (95%) in private practice settings (51%). The survey respondents had a mean of 13.6 years of post degree clinical experience, and spent an average of 45% of their clinical time working with adolescents. Survey results reveal a substantial similarity in test usage between the 1991 survey and the current investigation. For example, the Wechsler Intelligence Scales, Rorschach, Thematic Apperception Test (TAT), and Minnesota Multiphasic Personality Inventory (MMPI) remain among the widely used tests with adolescents. However, several changes were also noted including a reduction in the use of the Bender-Gestalt and increases in the use of parent and teacher rating instruments. The current findings are used to estimate the relative popularity of an extensive list of test instruments, compare current findings to 1991 survey results, and to examine several issues related to general effects of managed care procedures and policies on test usage with adolescents. PMID- 11037391 TI - Second-order confirmatory factor analysis of the WAIS-III. Wechsler Adult Intelligence Scale. AB - According to Vernon's structure-of-intellect paradigm, abilities can be conceptualized as a hierarchy, with a factor of general intelligence at the top of the hierarchy, and successively more specific abilities toward the bottom. This paradigm has proven useful for interpreting a number of Wechsler intelligence scales. However, most of the research with this paradigm has used exploratory factor analysis, and the validity of the paradigm for the newest Wechsler Adult Intelligence Scale (WAIS-III) has yet to be evaluated. The present study examined the WAIS-III using second-order confirmatory factor analysis, which is a more appropriate analytic tool when specific hypotheses are tested. Using the standardization sample for the WAIS-III (N = 2,450), support was found for the hierarchical factor structure with a second-order factor of general intelligence and four first-order factors of Verbal Comprehension, Perceptual Organization, Working Memory, and Processing Speed. PMID- 11037392 TI - Psychometric analysis of racial differences on the Maudsley Obsessional Compulsive Inventory. AB - Black university students scored significantly higher than White students on the Maudsley Obsessional Compulsive Inventory (MOCI). They tended to endorse more Cleaning and Checking subscale items in the pathological direction. Subsequent analyses examined whether this finding is a reflection of valid group differences in the prevalence of OCD or a psychometric artifact. Structured interviews were conducted to determine the correspondence of MOCI scores with OCD diagnoses. The race difference in endorsement frequency on the MOCI did not extend to OCD diagnoses. The MOCI scores showed modest predictive validity in Whites, but they did not predict interview-based diagnoses in Blacks. Multivariate item response theory was then employed to examine race differences in the Cleaning and Checking subscales. Equivalent item discrimination parameters fit the data for Black and White participants for both subscales. A more restrictive model in which relative item difficulties were also constrained to be equal for Black and White participants did not fit. This interaction between race and item difficulty suggests that the items do not have equivalent psychometric properties in Blacks and Whites. PMID- 11037393 TI - The diagnostic efficiency of the Rorschach Depression Index and the Schizophrenia Index: a review. AB - This review focuses on the diagnostic efficiency of the new versions of the Rorschach Comprehensive System Depression Index (DEPI) and the Schizophrenia Index (SCZI). Clinical diagnosis according to the Diagnostic and Statistical Manual of Mental Disorders was chosen as the external validation criteria. The sensitivity, specificity, and overall classification rates for the indices were presented from the studies or computed from the data when possible. The positive and negative predictive validity was estimated at three different base rates. As regards the DEPI the results showed a large variation in diagnostic performance as the index seemed to have relatively more success in identifying nonpsychotic and unipolar depression than psychotic and bipolar depression. The DEPI did not successfully identify depression among adolescent patients. As regards the SCZI the results more consistently indicated that the index effectively discriminates between psychotic and nonpsychotic patients and the predictive validity of both a positive and negative SCZI was found to be high. PMID- 11037394 TI - An investigation of the relationship between psychopathic traits and malingering on the psychopathic personality inventory. AB - This study employed a repeated-measures simulation design to examine (a) the specific effects of malingering on a recently developed measure of psychopathy, the Psychopathic Personality Inventory (PPI), and (b) the broader association between psychopathic traits and dissimulation. One hundred and forty-three participants completed the PPI twice (both under standard instructions and with instructions to feign psychosis), and also completed post-test questionnaires assessing their attitudes toward engaging in malingering across several hypothetical settings. When attempting to feign psychosis, participants produced elevated scores on a validity scale designed to identify deviant responding, and use of a cross-validated cutoff score with this scale produced high sensitivity and specificity rates across the honest and malingering conditions. Furthermore, PPI scores (in the honest condition) were significantly correlated with a willingness to engage in dissimulation across various hypothetical forensic/correctional scenarios. Results are discussed in terms of the "fakability" of the PPI, as well as the relationship between psychopathic personality features and malingering more generally. PMID- 11037395 TI - Repeated assessment of the Tower of Hanoi test: reliability and age effects. AB - The purpose of this research was to analyze the effects of repeating an executive function test. Three versions of the Tower of Hanoi (TOH) test were repeated three times each, with test-retest intervals of 2 months. Two groups of children participated in the research (7.7 and 11.6 years, n = 22 and n = 28). Repeating the assessment improved the performance and decreased the total performance time in both of the groups. The older participants improved their performance faster than the younger ones. The reliability of all the scores, besides the error scores, seemed to be satisfactory after the first few assessments. The stability of the scores was maintained through all the assessments. The planning time did not explain the variations of the achieved score. The reasons for the initially low reliabilities are discussed, and modifications for the test administration and scoring are suggested. PMID- 11037396 TI - Suggestions for revised scoring of the Tower of Hanoi test. AB - Detailed time and error analyses of the Tower of Hanoi (TOH) test was performed using four repeated assessments of eight children (ages 9-12 years), who had perceptual and problem solving deficits. The time before each move was measured. In addition to the traditionally counted time scores, new, relative time scores were computed in order to separate the planning time from the general reaction speed. New error scores were defined and sum scores of serious errors (perserative moves, illegal moves, and wrong results) and mild errors (self corrected moves, almost performed moves, and interrupted trials) were computed. The relative planning time correlated positively with the achieved score, and negatively with the serious errors. The serious errors correlated negatively with the achieved score. The relative planning time seems to measure the quality of planning better than does the raw planning time, and it is a recommended score for TOH analysis. The value of new error scores requires additional research. PMID- 11037397 TI - Acute chest pain--suspected aortic dissection. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037398 TI - Acute chest pain--suspected myocardial ischemia. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037399 TI - Acute chest pain--suspected pulmonary embolism. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037400 TI - Shortness of breath--suspected cardiac origin. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037401 TI - Chronic chest pain--suspected cardiac origin. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037402 TI - Blunt chest trauma--suspected aortic injury. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037403 TI - Blunt abdominal or pelvic trauma--suspected vascular injury. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037404 TI - Suspected lower extremity deep vein thrombosis. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037405 TI - Pulsatile abdominal mass. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037406 TI - Diagnostic imaging in patients with claudication. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037407 TI - Suspected congenital heart disease in the adult. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037408 TI - Suspected bacterial endocarditis. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037409 TI - Acute chest pain--no ECG evidence of myocardial ischemia/infarction. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037410 TI - Chronic chest pain without evidence of myocardial ischemia/infarction. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037411 TI - Recurrent symptoms following lower extremity arterial bypass surgery. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037412 TI - Recurrent symptoms following lower extremity angioplasty: claudication and threatened limb. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037413 TI - Sudden onset of cold, painful leg. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037414 TI - Unilateral upper extremity swelling and pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037415 TI - Hematemesis. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037416 TI - The patient with suspected small bowel obstruction: imaging strategies. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037417 TI - Imaging strategies in the initial evaluation of the jaundiced patient. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037418 TI - Pre-treatment staging of colorectal cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037419 TI - Imaging of blunt abdominal trauma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037420 TI - Evaluation of patients with acute right upper quadrant pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037421 TI - Evaluation of acute right lower quadrant pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037422 TI - Evaluation of left lower quadrant pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037423 TI - Suspected abdominal abscess. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037424 TI - Imaging recommendations for patients with newly suspected Crohn's disease, and in patients with known Crohn's disease and acute exacerbation or suspected complications. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037425 TI - Liver lesion characterization. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037426 TI - Imaging evaluation of the palpable abdominal mass. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037427 TI - Acute pancreatitis. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037428 TI - Imaging evaluation of patients with acute abdominal pain and fever. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037429 TI - Suspected liver metastases. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037430 TI - Imaging recommendations for patients with dysphagia. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037431 TI - Screening for colorectal cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037432 TI - Imaging evaluation of suspected ankle fractures. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037433 TI - Cervical spine trauma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037434 TI - Diagnostic imaging of avascular necrosis of the hip. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037435 TI - Soft tissue masses. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037436 TI - Bone tumors. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037437 TI - Stress/insufficiency fractures (excluding vertebral). American College of Radiology. ACR Appropriateness Criteria. PMID- 11037438 TI - Imaging of the multiply injured patient. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037439 TI - Metastatic bone disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037440 TI - Evaluation of the patient with painful hip or knee arthroplasty. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037441 TI - Shoulder trauma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037442 TI - Imaging diagnosis of osteomyelitis in patients with diabetes mellitus. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037443 TI - Nontraumatic knee pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037444 TI - Chronic ankle pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037445 TI - Chronic wrist pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037446 TI - Chronic elbow pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037447 TI - Chronic neck pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037448 TI - Chronic foot pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037449 TI - Acute trauma to the knee. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037450 TI - Acute hand and wrist trauma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037451 TI - Follow-up examinations for bone tumors, soft tissue tumors, and suspected metastasis post therapy. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037452 TI - Hip arthroplasty--radiography procedure recommendations. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037453 TI - Chronic hip pain. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037454 TI - Osteoporosis and bone-mass measurement. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037455 TI - Cerebrovascular disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037456 TI - Progressive neurological deficit. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037457 TI - Epilepsy. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037458 TI - Vertigo and hearing loss. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037459 TI - Acute low back pain--radiculopathy. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037460 TI - Atraumatic isolated headache--when to image. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037461 TI - Myelopathy. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037462 TI - Head trauma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037463 TI - Dementia. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037464 TI - Imaging of intracranial infections. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037465 TI - Multiple sclerosis--when and how to image. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037466 TI - Neuroendocrine imaging. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037467 TI - Ataxia. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037468 TI - Orbits, vision, and visual loss. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037469 TI - Spine trauma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037470 TI - Neurodegenerative disorders. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037471 TI - Work-up of the solitary pulmonary nodule. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037473 TI - Routine daily portable x-ray. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037472 TI - Staging of bronchogenic carcinoma, non-small cell lung carcinoma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037474 TI - Routine chest radiographs in uncomplicated hypertension. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037475 TI - Hemoptysis. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037476 TI - Rib fractures. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037477 TI - Dyspnea. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037478 TI - Acute respiratory illness. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037479 TI - Acute respiratory illness in HIV-positive patients. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037480 TI - Screening for pulmonary metastases. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037481 TI - Radiologic investigation of patients with renovascular hypertension. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037482 TI - Recurrent lower urinary tract infections in women. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037483 TI - Imaging in acute pyelonephritis. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037484 TI - Acute onset flank pain, suspicion of stone disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037485 TI - Radiologic investigation of patients with hematuria. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037486 TI - Obstructive voiding symptoms secondary to prostate disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037487 TI - Pretreatment staging of invasive transitional cell carcinoma of the bladder. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037488 TI - Pretreatment staging of clinically localized prostate cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037489 TI - Acute onset of scrotal pain (without trauma, without antecedent mass). American College of Radiology. ACR Appropriateness Criteria. PMID- 11037490 TI - Radiologic investigation of causes of renal failure. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037491 TI - Renal cell carcinoma staging. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037492 TI - Diagnostic approach to renal trauma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037493 TI - Trauma to the bladder and urethra. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037494 TI - Staging of testicular malignancy. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037495 TI - Indeterminate renal masses. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037496 TI - The incidentally discovered adrenal mass. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037497 TI - Follow-up of renal cell carcinoma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037498 TI - Follow-up imaging of bladder carcinoma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037499 TI - Post-treatment follow-up of prostate cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037500 TI - Vomiting in infants up to 3 months of age. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037501 TI - Imaging of the pediatric patient with seizures. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037502 TI - The limping child. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037503 TI - Imaging the child with suspected physical abuse. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037504 TI - Sinusitis in the pediatric population. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037505 TI - Developmental dysplasia of the hip. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037506 TI - Fever without source. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037507 TI - Imaging evaluation of acute right lower quadrant and pelvic pain in adolescent girls. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037508 TI - Hematuria. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037509 TI - Urinary tract infection. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037510 TI - Headache. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037511 TI - Ovarian cancer screening. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037512 TI - Role of imaging in abnormal vaginal bleeding. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037513 TI - First trimester bleeding. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037514 TI - Role of imaging in second and third trimester bleeding. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037515 TI - Staging and follow-up of ovarian cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037516 TI - Evaluation of multiple gestations. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037517 TI - Growth disturbances: risk of intrauterine growth retardation. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037518 TI - Role of imaging in cancer of the cervix. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037519 TI - Suspected adnexal masses. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037520 TI - Premature cervical dilatation. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037521 TI - Endometrial cancer of the uterus. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037522 TI - Imaging work-up for stage I breast carcinoma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037523 TI - Appropriate imaging work-up of palpable breast masses. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037524 TI - Work-up of nonpalpable breast masses. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037525 TI - Appropriate imaging work-up of breast microcalcifications. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037526 TI - Inferior vena cava filter placement. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037527 TI - Iliac angioplasty. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037528 TI - Percutaneous transluminal renal angioplasty. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037529 TI - Needle biopsy in the thorax. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037530 TI - Thrombolysis for lower extremity arterial and graft occlusions. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037531 TI - Percutaneous biliary drainage in malignant biliary obstruction. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037532 TI - Percutaneous catheter drainage of infected intra-abdominal fluid collections. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037533 TI - Bone metastases. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037534 TI - Pre-irradiation evaluation and management of brain metastases. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037535 TI - Solitary brain metastasis. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037536 TI - Multiple brain metastases. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037537 TI - Follow-up and retreatment of brain metastasis. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037538 TI - Ductal carcinoma in situ and microinvasive disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037539 TI - Postmastectomy radiotherapy. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037540 TI - Locally advanced breast cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037542 TI - Conservative surgery and radiation in the treatment of stage I and II carcinoma of the breast. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037541 TI - Local regional recurrence and salvage surgery. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037543 TI - Staging evaluation for patients with Hodgkin's disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037544 TI - Pediatric Hodgkin's disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037545 TI - Stage III and IV Hodgkin's disease treatment guidelines. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037546 TI - Follow-up of Hodgkin's disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037547 TI - Staging of non-small cell lung carcinoma. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037548 TI - Postoperative radiotherapy in non-small cell lung cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037549 TI - Non-small cell lung cancer, nonsurgical, aggressive therapy. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037550 TI - Neoadjuvant therapy for marginally resectable (clinical N2), non-small cell lung cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037551 TI - Non-aggressive, non-surgical treatment of inoperable non-small cell lung cancer (NSCLC). American College of Radiology. ACR Appropriateness Criteria. PMID- 11037552 TI - Follow-up of non-small cell lung cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037553 TI - Staging evaluation for patients with adenocarcinoma of the prostate. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037554 TI - Permanent source brachytherapy for prostate cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037555 TI - Locally advanced (high-risk) prostate cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037556 TI - Node-positive prostate cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037557 TI - Postradical prostatectomy irradiation in carcinoma of the prostate. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037558 TI - Treatment planning for clinically localized prostate cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037559 TI - Definitive external beam irradiation in stage T1 and T2 carcinoma of the prostate. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037560 TI - Management of resectable rectal cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037561 TI - Locally unresectable rectal cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037562 TI - Rectal cancer: presentation with metastatic and locally advanced disease. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037563 TI - Anal cancer. American College of Radiology. ACR Appropriateness Criteria. PMID- 11037564 TI - Amphetamine et al. PMID- 11037565 TI - Medication errors. PMID- 11037566 TI - Telling the truth about cancer: views of elderly patients and their relatives. AB - The aim of this study was to compare the attitudes of elderly patients and their relatives towards telling the truth about cancer. 120 patients were asked if they would wish to be told about bad news, such as cancer, which might emerge during the admission. Matched relatives were asked if such information should be disclosed to the patient. Of the 120 patients, 99 (83%) wanted to be told the truth; 66 relatives (55%) relatives wanted their next of kin informed. There was agreement in 73 (61%) pairs. The kappa statistic was 0.16 (95% confidence interval -0.03 to 0.35), which indicates poor agreement. We conclude that most elderly people wish to be informed of a diagnosis of cancer. Patient preferences cannot be predicted by talking to relatives. PMID- 11037567 TI - Secondary prevention following fractured neck of femur: a survey of orthopaedic surgeons practice. AB - Fractured neck of femur is one of the most common reasons for hospital admission in the elderly population. Despite this little attention is given to the area of secondary prevention. To assess the variation in clinical practice among orthopaedic surgeons with regard to the use of secondary preventive measures in relation to fractured neck of femur. A postal survey using a self-completed questionnaire among all orthopaedic surgeons throughout the Republic of Ireland. The study questionnaire addressed unit information prophylaxis for deep venous thrombosis and investigation and secondary prevention of fractured neck of femur. It gave a clinical scenario of a 72-year old female who was admitted with a fractured neck of femur after a minor fall. A reminder was sent to the non responders. Data were analysed using chi-squared test. The number of completed questionnaires returned was 89 giving a response rate of 66%. Of these 71(82.6%) reported that they would not initiate or recommend investigation of the extent of the underlying osteoporosis in the specific case upon which the questionnaire was based. With regard to the issue of secondary prevention 16(18%) stated that they would prescribe Vitamin D and 26(29%) would prescribe calcium supplements. 15(17%) would use bisphosphonates and 15(17%) would prescribe hormone replacement therapy. Only one respondent would use an external hip protector. There is an underuse of secondary preventive measures with regard to fractured neck of femur. There needs to be clear definition of roles and perhaps the development of agreed local protocols in order that patients with hip fractures do not miss out on potentially beneficial treatment. There is an underuse of secondary preventive measures with regard to fractured neck of femur. There needs to be clear definition of roles and perhaps the development of agreed local protocols in order that patients with hip fractures do not miss out on potentially beneficial treatment. PMID- 11037568 TI - Quality of life following total hip replacement. AB - Orthopaedic surgeons have traditionally used disease-specific scoring systems that focus on relief of pain and improvement in function. However, these allow little scope for patient assessed outcomes such as quality of life. This study assessed quality of life before and after Total Hip Replacement (THR) using the Short Form 36 (SF-36) questionnaire. 100 patients were interviewed pre operatively and 9 months to one year post-operatively. Data on waiting times/length of stay/complication rates were collected by reviewing clinical notes. The mean length of stay was 22.0 +/- 10.6 days, with a pre-operative length of stay of 5.0 days. 12 patients in all had a post-operative complication, including one post-operative death. There was a statistically significant improvement in overall SF-36 score (+19%) and in 7 out of the 8 parameters that make up the SF-36 score (p < 0.001). Patients undergoing THR in Navan showed a statistically significant improvement in their quality of life post-operatively. Complication rates were in keeping with international norms. It is apparent that patients receive a high quality of care. PMID- 11037569 TI - Renal transplantation in pre-school children. AB - This short report describes the outcome of 13 renal transplants in 11 children under 5 years of age. Nine (82%) of the 11 recipients are alive; 2 children died with functioning grafts. Approximately 50% of grafts are functioning at 5 years post transplantation. Children with congenital kidney malformations can be successfully managed to transplantation. PMID- 11037570 TI - Evaluating practice at an Irish tertiary level fertility clinic 1995/98. AB - This paper audits patients patterns and the diagnostic activities of the Rotunda Hospital, Infertility Clinic by following 186 new patients first attending between 1st March 1995 and 28th February 1996 and their progress in the clinic through to 28th February 1998. Sixty-four percent of patients came from the Eastern Health Board. Sixty-nine per cent had been previously investigated elsewhere. The non-notified drop-out rate was 23%. Of the 158 patients fully investigated, the male partner was the major factor in 27% of couples. Tubal problems accounted for 20% and endometriosis 13%. An ovulation factor was found in 13%. Twenty-seven percent of couples were given the label of unexplained infertility. In 8% more than one factor was present. Fifty-five percent were referred on for IVF/ICSI. Many at this tertiary referral centre had already attended elsewhere. A very high proportion of these had repeat investigations to complete the fertility profile. This policy is questioned as it is not cost effective. The incidence of cause of infertility in the main has little changed in 15 years. PMID- 11037571 TI - A review of the epidemiology of leptospirosis in the Republic of Ireland. PMID- 11037572 TI - Hashimoto's encephalopathy; an unusual cause of status epilepticus. PMID- 11037573 TI - Massive retroperitoneal liposarcoma. PMID- 11037574 TI - Occupational contact allergy to immersion oil. PMID- 11037575 TI - Hospital cost of acute myocardial infarction in the thrombolytic era. PMID- 11037576 TI - Residential randon exposure and lung cancer risk in Misasa, Japan: a case-control study. AB - In order to investigate an association between residential radon exposure and risk of lung cancer, a case-control study was conducted in Misasa Town, Tottori Prefecture, Japan. The case series consisted of 28 people who had died of lung cancer in the years 1976-96 and 36 controls chosen randomly from the residents in 1976, matched by sex and year of birth. Individual residential radon concentrations were measured for 1 year with alpha track detectors. The average radon concentration was 46 Bq/m3 for cases and 51 Bq/m3 for controls. Compared to the level of 24 or less Bq/m3, the adjusted odds ratios of lung cancer associated with radon levels of 25-49, 50-99 and 100 or more Bq/m3, were 1.13 (95% confidence interval; 0.29-4.40), 1.23 (0.16-9.39) and 0.25 (0.03-2.33), respectively. None of the estimates showed statistical significance, due to small sample size. When the subjects were limited to only include residents of more than 30 years, the estimates did not change substantially. This study did not find that the risk pattern of lung cancer, possibly associated with residential radon exposure, in Misasa Town differed from patterns observed in other countries. PMID- 11037577 TI - Sensitization by wortmannin of heat- or X-ray induced cell death in cultured Chinese hamster V79 cells. AB - Here we found that wortmannin sensitized Chinese hamster V79 cells to hyperthermic treatment at 44.0 degrees C as determined either by colony formation assay or by dye exclusion assay. Wortmannin enhanced heat-induced cell death accompanying cleavage of poly (ADP-ribose) polymerases (PARP). Additionally, the induction of heat shock protein HSP70 was suppressed and delayed in wortmannin treated cells. Heat sensitizing effect of wortmannin was obvious at more than 5 or 10 microM of final concentrations, while radiosensitization was apparent at 5 microM. Requirement for high concentration of wortmannin, i.e., order of microM, suggests a possible role of certain protein kinases, such as DNA-PK and/or ATM among PI3-kinase family. The sensitization was minimal when wortmannin was added at the end of heat treatment. This was similar to the case of X-ray. Since heat induced cell death and PARP cleavage preceded HSP70 induction phenomenon, the sensitization to the hyperthermic treatment was considered mainly caused by enhanced apoptotic cell death rather than secondary to suppression or delay by wortmannin of HSP70 induction. Further, in the present system radiosensitization by wortmannin was also at least partly mediated through enhancement of apoptotic cell death. PMID- 11037578 TI - Novel approach to in vivo screening for radioprotective activity in whole mice: in vivo electron spin resonance study probing the redox reaction of nitroxyl. AB - Previously, we reported that X-irradiation enhanced the signal decay of a spin probe injected into whole mice measured by in vivo ESR, and that the observed enhancement was suppressed by the pre-administration of cysteamine, a radioprotector [Miura, Y., Anzai, K., Urano, S. and Ozawa, T. (1997) Free Rad. Biol. Med. 23: 533-540]. In the present study, the suppression activity of the X ray-induced increase in the ESR signal decay rate (termed suppression index, SI) was measured for several radioprotectors: 5-hydroxytryptamine (5-HT), S-2-(3 aminopropylamino)-ethylphosphorothioic acid (WR-2721), 4-hydroxy-2,2,6,6 tetramethyl-piperidine-N-oxyl (TEMPOL), cimetidine, interleukin-1 beta (IL-1 beta) and stem cell factor (SCF). The enhancement of the ESR signal decay of carbamoyl-PROXYL due to X-irradiation was suppressed by a treatment with all of the radioprotectors examined, showing positive SI values. However, a dose dependency of 5-HT or WR-2721 was not observed, suggesting that several mechanisms exist for radioprotection and a modification of the signal decay rate. Although the in vivo ESR system cannot be used in place of the 30-day survival method for the assessment of radioprotectors, this system might be applicable to in vivo, non-invasive screening prior to using the 30-day survival method. PMID- 11037579 TI - Dose and dose-rate effects of X rays and fission neutrons on lymphocyte apoptosis in p53(+/+) and p53(-/-) mice. AB - Following the exposure of mice to X rays or fission neutrons, the frequency (F) of apoptosis was measured after 4 h, and the weight loss or lymphocyte content loss in the thymus and spleen was measured after 24 h. In p53(+/+) mice, F increased linearly with the dose (D (Gy)) and the induced rate per Gy of F (detected by TUNEL staining) was 0.05 and 0.23 for X rays and fission neutrons, respectively. Therefore, the RBE of fission neutrons was 4.6 for apoptosis induction. This indicates that radiation-induced apoptosis is mostly due to double strand breaks (DSBs) in DNA because we previously obtained almost the same RBE value of fission neutrons for the induction of crossover mutations in Drosophila melanogaster, which arise from the recombinational repair of DSBs. In p53(+/+) mice, decreases in the organ weight and the lymphocyte content were observed for the thymus and the spleen 24 h after X-irradiation. These atrophic changes in the thymus and the spleen quantitatively corresponded to the total apoptotic cell deaths occurring in them. However, in p53(-/-) mice, no vigorous apoptosis was induced after X-irradiation, and hyperplastic changes in the weight and the lymphocyte content appeared in the thymus and the spleen 24 h after X irradiation. In p53(+/+) mice, there was no difference in the induced rate per Gy of reduction in the surviving fraction of lymphocytes between acute (0.4 Gy/min) and chronic (3 mGy/min) gamma-irradiations. Namely, radiation-induced apoptosis in lymphocytes is a dose-rate independent event. PMID- 11037580 TI - Application of a newly developed photoluminescence glass dosimeter for measuring the absorbed dose in individual mice exposed to low-dose rate 137Cs gamma-rays. AB - A photoluminescence glass dosimeter, GD-301, was applied to the measurement of low absorbed doses in mice exposed to low-dose rate 137Cs gamma-rays. The dosimeter system consists of small rod-shaped glass chip detectors capable of embedded in the body of a mouse and an automatic readout device equipped with a standard detector irradiated with 137Cs gamma-source. The measured absorbed doses were compared with the "exposure" estimated by an ionization chamber and with the doses measured by a BeO:Na thermoluminescence system. The results clearly demonstrate the superiority of the glass dosimetry regarding simplicity of operation, stability of long-term dose accumulation and good detector uniformity, which allow accurate tissue dosimetry. PMID- 11037581 TI - Quantitative detection of apoptotic thymocytes in low-dose X-irradiated mice by an anti-single-stranded DNA antibody. AB - The quantitative detection of apoptotic cells in low frequency in the thymus of mice irradiated with X-rays using an anti-single-stranded DNA antibody was explored. The antibody against single-stranded DNA (anti-ssDNA) was obtained with rabbits hyperimmunized with complexes of alkaline-denatured calf thymus DNA. AKR female mice were irradiated with 10 to 100 cGy or 4 Gy X-rays; thereafter, thymus sections were prepared at various times after irradiation. The detection and counting of apoptotic cells in the section were performed after histochemical staining using an anti-ssDNA antibody. The results demonstrate that, although sensitive and quantitative detection of apoptotic cells in irradiated thymus using the anti-ssDNA antibody is possible, the sensitivity is lower compared to that of in situ end-labeling methods, such as TUNEL or ISEL. The antibodies could also be used for rat thymus and spleen. In addition, an increase in positively stained cells by both methods was detected as early as 6 min after the irradiation of mice. PMID- 11037582 TI - Relative biological effectiveness of carbon ions for causing fatal liver failure after partial hepatectomy in mice. AB - To evaluate the acute phase damage to liver by carbon ions, BALB/c mice were irradiated with carbon ions or X-rays after two-thirds partial hepatectomy, and their survival was followed. The 50% lethal dose within 60 days (LD50/60) was 42.2 +/- 0.25 Gy (standard error) for X-rays, and 22.7 +/- 0.25 Gy for carbon ions. The relative biological effectiveness (RBE) of carbon ions was 1.86 (95% confident limits: 1.69-2.04) as calculated from the LD50/60. Mice irradiated at much higher doses, 60 Gy of X-rays or 24 Gy of carbon ions, showed significantly higher serum ammonia levels and lower serum albumin levels than normal, suggesting hepatic failure as a cause of death. Hepatocytes showed karyorrhexis and karyolysis in carbon ion irradiated and spotty necrosis in X-ray irradiated mice, suggesting nuclear damage. Mice irradiated with LD50 of X-rays or carbon ions had a remarkably lower bromodeoxyuridine (BrdU) labeling index and mitotic index than control. Treatments with both BrdU and vincristine showed that none of the hepatocytes that synthesized DNA after irradiation completed mitosis, indicating G2 arrest. The liver weight of irradiated mice significantly decreased depending on the dose. Carbon ions as well as X-rays damaged hepatocytes directly and suppressed liver regeneration leading to fatal liver failure. PMID- 11037584 TI - [Cheers for the randomized controlled trial! The only method able to evaluate benefit of medical measures]. PMID- 11037583 TI - LET dependency of heavy-ion induced apoptosis in V79 cells. AB - We investigated the relationship between the LET values and cell death, defined as either apoptosis or loss of reproductive integrity (reproductive death), using Chinese hamster V79 cells. The cells were irradiated with X-rays or carbon-ion beams from the Heavy Ion Medical Accelerator in Chiba (HIMAC) at the National Institute of Radiological Sciences (NIRS). Apoptosis was defined based on the morphological change upon treating of cells with caffeine. The apoptotic index, the ratio of apoptotic cells to the total, after exposure to 2 Gy of X-rays was 0.043. Upon irradiation with carbon-ion beams, the index was gradually increased with increasing LET values, reaching a maximum of 0.076 at 110 keV/micron, and then decreased to 0.054 at 237 keV/micron. An analogous pattern of the LET dependence was observed between reproductive death and apoptotic death. The cell survival values obtained after 2 Gy exposure (SF2) were 0.64, 0.13, and 0.24, respectively. A similar trend was found for the RBE values calculated from the initial slope for both apoptosis and reproductive death. These results strongly suggest that the target for both types of cell death is the same. PMID- 11037585 TI - [Duplicate publication a way of embellishing research results. Unethical misuse which threatens the validity of systematic reviews and meta-analysis]. AB - Studies showing favourable results tend to be published more than once. This practice of duplicate publication is, however, dishonest and unethical on the part of scientists and pharmaceutical companies. Inclusion into a meta-analysis of the results from multiple papers derived from one and the same controlled study gives an inflated picture of the efficacy of the treatment. If manufacturers and researchers not only made all their study data public but also registered trials prospectively these pitfalls could be avoided. PMID- 11037586 TI - [Laparoscopic surgery--evidence-based ?]. AB - The literature has been searched for current results in laparoscopic cholecystectomy, hernia repair, appendectomy and fundoplication. This was performed as a systematic review. Laparoscopic cholecystectomy was judged to be safe and cost/effective, with good patient acceptability. However a need for further studies is indicated. Laparoscopic technique in hernia repair has a longer learning curve and is more expensive than open repair, with no major difference in recurrence rates. It is preferable in bilateral repairs. Laparoscopic appendectomy in the hands of experienced surgeons is cost/effective. Time to recovery is shorter and the rate of infectious complications is lower than in conventional procedures. There are still too few results reported from laparoscopic fundoplication to permit reliable conclusions. PMID- 11037587 TI - [Evidence based complementary medicine--does it exist? The first international congress discussed value, safety and risks]. PMID- 11037588 TI - [Treatment of advanced prostatic cancer]. AB - In advanced prostate cancer when the tumor has metastasized, endocrine therapy is the primary treatment alternative. Endocrine therapy was introduced 60 years ago by Huggins and Hodges in the form of castration or estrogen treatment. Unfortunately we must now concede that no decisive breakthrough in the pharmacological treatment of prostate cancer has occurred since that time. We do have several different endocrine treatment alternatives at our disposal today- all with varying advantages and drawbacks. We do, however, eagerly await new therapeutic options for the pharmacological eradication of prostatic adenocarcinoma. Today our interest is focused on methods of tailoring endocrine therapy in ways that offer the individual patient an optimal combination of quality of life and anticancer efficacy. The various opinions and research results which inform current strategies of endocrine therapy are described briefly. PMID- 11037590 TI - [Diet probably plays an important role in the development of prostatic cancer]. AB - Prostate cancer is the most common malignant disease in Sweden and the most common cause of cancer-related death among Swedish men. There is, however, a wide geographical variation in the age-standardized incidence and mortality rates. The highest incidence is found in north-western Europe and the US and the lowest in the Asian countries. The reasons for these discrepancies are thought to be related to environmental factors such as variations in dietary pattern. High intake of calories, high Body Mass Index, and consumption of animal fat are all associated with an increased risk of prostate cancer, while high intake of soy and other phytoestrogens, selenium, vitamin A and high serum levels of vitamin D are associated with low risk. As well, gonadal hormones and growth factors are believed to be involved in the complex etiology of prostate cancer. Genetic factors play an important role in the development of prostate cancer, and a hereditary form of the disease, accounting for approximately 5-10% of cases, has been identified. In order to develop effective preventive strategies to reduce prostate cancer mortality and morbidity, it is necessary to expand our knowledge about the etiology of this common disease. PMID- 11037589 TI - [Brachytherapy with palladium-103--a new way to deliver radiation therapy for localized prostatic cancer]. AB - The incidence of prostate cancer is increasing and, due to the use of PSA, more and more patients are diagnosed with localized disease. These patients may be offered treatment with the intent to cure. Traditionally, this has been either by radical prostatectomy or some form of radiation therapy. This article describes the minimally invasive method with permanent seed implantation. The procedure, indications, contraindications, side effects and results from the literature 28 patients have been treated so far with acceptable proximate side effects and reductions in serum-PSA as expected. PMID- 11037591 TI - [HIV epidemics without mercy. Several generations of children and adolescents in many African countries will be eradicated]. PMID- 11037592 TI - [Prevention of perinatal asphyxia. Can more be done by fetal monitoring?]. AB - Intrapartum death has become a rare event since the introduction of electronic fetal monitoring (EFM). Randomized studies of EFM versus intermittent auscultation show that EFM reduces the risk of low Apgar score at birth and of neonatal seizures, although it may be possible to ensure low perinatal mortality even with intermittent auscultation alone. The rate of operative deliveries may increase with EFM, at least if scalp blood sampling is not used. Since scalp blood sampling is an invasive procedure providing only temporary information, the search goes on for new methods as complements to EFM. Fetal pulse oximetry and ST analysis of fetal ECG (STAN) are two methods currently being tested in clinical studies. Regardless of the efficiency of monitoring methods that we may develop, there will always be a need for competent staff at our labor wards. Ongoing training of staff is necessary if asphyxia is to be avoided as far as possible. PMID- 11037593 TI - [Telemedicine for home-based monitoring of newborn infants reduces the length of care and increases the quality. Pilot projects for newborn infants with chronic lung disease in need of oxygen therapy]. PMID- 11037594 TI - [School maladjustment common among children with very low birth weight Special attention and support are required during school start]. AB - Children of very low birth weight (VLBW), defined as less than 1500 g, and normal birth weight controls (NBW) were enrolled in a long-term follow-up study. Five of 86 surviving VLBW children had a neurological handicap. Seventy VLBW children and 72 NBW children were re-examined at the age of nine, which entailed a neurological examination, a non-verbal intelligence test and a test for reading ability, mathematical skills and vocabulary. Their behavior was rated regarding hyperactivity, social behavior and fine and motor skills. The two groups differed with regard to the neurological examination and the tests, with poorer results shown for the low birth weight group. The VLBW children were also more hyperactive and scored lower on fine motor skills. Considering only those who scored normally as regards non-verbal intelligence (54% of VLBW children and 88% of controls), practically all differences disappeared. PMID- 11037595 TI - [The debate goes on: deficient stringency in MFR's documentation on child health care]. PMID- 11037596 TI - [The MFR documentation on child health care: more guidelines are required when it comes to children with developmental deviations]. PMID- 11037597 TI - [Costs and problems in cervix cancer screening are considerably underestimated]. PMID- 11037598 TI - [Reply to a comment: Why did the diagnosis fibromyalgia first appear at the end of the 20th century?]. PMID- 11037599 TI - [The scientist as a patient: the one who knows where the shoe pinches]. PMID- 11037600 TI - [Natriuretic peptides in the diagnosis of heart symptoms]. PMID- 11037601 TI - [Serious alternative medicine can be a complement]. PMID- 11037602 TI - [A dialogue depends on mutual respect]. PMID- 11037603 TI - [On psychiatric diagnoses--a study from Gothenburg]. PMID- 11037604 TI - [What is evidence-based treatment of otitis?]. PMID- 11037605 TI - [DSM diagnostics in psychiatry]. PMID- 11037606 TI - [Risks with mobile phones--blocking mechanisms?]. PMID- 11037607 TI - [Wrong conclusion on hospital-based home care services, SBU!]. PMID- 11037608 TI - [Johann Sebastian Bach's mortal remains found and identified after an assidious detective work]. PMID- 11037609 TI - [Perinatal origin of malignant diseases]. AB - In the present days the molecular genetical investigations are the mainstream of establishing the etiology of malignant diseases. Beside these surveys, prenatal, neonatal, environmental and developmental risk factors for malignancy have repeatedly been investigated during the last few years. Mounting evidences show the importance of the intrauterine and perinatal period in tumor genesis and health quality in later life. This review article summarizes the results of traditional epidemiologic studies which identified a number of risk factors for malignancy. These easily detectable anamnestic data, developmental, physical features can further support the prenatal origin of tumors and can give new directions for the modern molecular genetical investigations. PMID- 11037610 TI - [Effect of fructose-1,6-diphosphate on myocardial purin and pyrimidin metabolism during coronary artery bypass grafting surgery]. AB - During ischaemia, the glycolytic pathway (Embden-Meyerhof) is up regulated in an attempt to produce ATP anaerobically. However, this is short-lived due to negative feedback on the key glycolytic enzyme phosphofructokinase by accumulating lactate. Fructose-1,6-diphosphate (FDP), a high energy intermediary metabolite of this pathway, is unique in that is enters glycolysis distal to this inhibitory site. Exogenously administered FDP should therefore theoretically yield ATP independent of lactate accumulation and thereby ameliorate ischaemic injury. Clinical benefit has been shown in coronary artery bypass grafting (CABG) surgery, congestive cardiac failure and adult respiratory distress syndrome. Ischaemia-reperfusion injury induced by cardiopulmonary bypass (CPB) presents clinically as an impairment of myocardial function in the postoperative period. At a cellular level this reflects myocardial metabolic changes and nucleotide degradation (directly linked to high energy phosphate turnover). Quantification of myocardial nucleotide catabolite release therefore provides useful information regarding intermediary metabolism and cytoprotection conferred to myocardial (inosine, uridine) and endothelial (hypoxanthine) tissue. The authors investigated the myocardial cytoprotective effects of FDP in 16 patients scheduled for elective CABG surgery. Aortic and coronary sinus blood were collected directly into liquid nitrogen and analysed by high performance liquid chromatography prior to CPB and at different time points after reperfusion. FDP was administered intravenously in 8 patients and 5% dextrose was administered in 8 other patients. Analysis of transmyocardial (coronary sinus-aortic) nucleotide metabolite levels showed increased release of inosine, hypoxanthine and uridine in both the FDP and the control groups following reperfusion. However, compared to baseline (pre-aortic clamping) values, hypoxanthine and inosine concentrations were significantly elevated at 0, 1, 5 and 10 minutes following reperfusion in the control group. This was in contrast to earlier recovery to baseline levels (after 5 minutes of reperfusion) in the FDP group. Furthermore, when compared to control group, the hypoxanthine and inosine concentrations were significantly decreased by FDP treatment. Uridine concentrations were significantly elevated at 1 and 5 minutes in the control group and no significant change was observed in the FDP group. In conclusion, these data suggest that FDP, through an intermediary metabolic effect, may contribute to myocardial and endothelial cytoprotection during the ischaemic insult of cardiac surgery. PMID- 11037611 TI - [Cytoprotective effect of Amifostine in the combined chemotherapy of advanced ovarian carcinoma]. AB - The authors treated 10 patients with advanced (stage III-IV) ovarian cancer with a high-dose cyclophosphamide and cisplatin (CP) combined therapy in amifostine protection. Due to the effect of the cytoprotective agent the cycles of treatment were well tolerated and 7 of 10 patients achieved a complete remission. Nephrotoxicity, allergic reaction or minor complications did not occur. Starting of the next treatment cycle was to be delayed in only one case because of anaemia. The agent has an implicit place in the treatment of patients with ovarian cancer in order to achieve better results and improve the quality of life. PMID- 11037612 TI - [Are the hemochromatosis (HFE) gene mutation and hepatitis C virus (HCV) infection risk factors for porphyria cutanea tarda?]. AB - As it is not clear whether mutations in hemochromatosis gene (HFE) and hepatitis C virus (HCV) act independently in the pathogenesis of porphyria cutanea tarda (PCT), and prevalence of both risk factors reveals a great variety in different parts of the world, PCT patients from our Central East European country were investigated for this aspect. The occurrence of the C282Y and H63D mutations in HFE gene were determined in 19 PCT patients and compared with the reported control frequencies. Furthermore, the presence of HCV infection was determined and related to the patients' HFE status. The C282Y mutation was found in 3/19 cases (one patient was homozygous and two heterozygous), with an 10.5% allele frequency (vs. 3.8% control) (p < 0.05). Five patients were heterozygous for the H63D mutation, allele frequency 13.1%, which did not differ from the reported control prevalence of 12.3%. Six patients (31.7%) were HCV-RNA positive, out of the six one was heterozygous for H63D mutation and one was compound heterozygous. HCV infection and HFE C282Y mutations may probably be independent predisposing factors for development of PCT in Hungarian patients. PMID- 11037613 TI - [Combined chemotherapy after prolonged oral etoposide maintenance therapy in patients with small-cell lung cancer]. AB - The authors have reported the results of induction, and maintenance therapy of 36 patients suffering from small-cell lung cancer. The induction chemotherapy with cisplatin and etoposide proved to be successful in 34 cases out of the 36 patients. The patients were divided in two groups following the induction treatment: with 17 patients no further chemotherapy happened. The other 17 patients received maintenance cytotoxic treatment with prolonged oral etoposide. There was a significant increase in the average survival time in cases of patients who received maintenance therapy (p < 0.001). The average survival of these patients was 908.4 days, while that of the patients of the control group proved to be 404.5 days. PMID- 11037614 TI - [Index Copernicus: The Central and Eastern European Journals Ranking System. Why indexing needed in the region?] . AB - Index Copernicus is ranking system set up by members of the medical community in the Region. There were created five groups of parameters like scientific, editorial and technical quality, circulation and frequency-market stability, which allow for the generation of such a ranking system. The Authors of the Ranking System are aware of the deficiencies of parametrical analysis of science, however they believe the numbers at least set up clear, objective and just rules for all. Index Copernicus could be said the primary objectives of the system for which it has been created for. PMID- 11037615 TI - [Scientometry of research publications]. PMID- 11037616 TI - [Guidelines for authors for complying with biometric methods in the content and form of their research reports] . PMID- 11037617 TI - [Autoimmune dermatoses: new therapeutic strategies]. AB - In the recent past, advances in both basic and clinical research have considerably contributed to the understanding of the cellular and molecular events that lead to autoimmune diseases. Nevertheless, current treatment protocols are essentially confined to systemic corticosteroids and cytotoxic agents. These are effective in controlling inflammation, however the occurrence of relapses after the treatment is stopped and the risks of toxicity and long term immunosuppression necessitate the search for more selective strategies of immunointervention. Major efforts are directed towards therapies with increased cellular selectivity and, ideally, autoantigen specificity. Cyclosporine A, monoclonal antibodies (anti-CD4) and immunotoxins (IL-2 fusion toxins) currently constitute the main clinical orientation. At the experimental level, strategies are directed towards antigen-specific manipulation of costimulatory pathways (anti-gp39, CTLA4Ig), peptide therapy (MHC antagonism, TCR vaccination), and tolerance induction to the autoantigen (oral tolerization). Alternatively, intervention in the cytokine cascade of the inflammatory response by means of cytokine-induced immunomodulation (IL-4, IL-10) or cytokine antagonism (anti-TNF alpha) are strategies with a therapeutic potential. However, it must be borne in mind, that in animal models of antigen-induced autoimmune disease, it is simple to intervene in the development of autoimmunity, since the induction events are designed by the investigator. In contrast, in the majority of patients with autoimmune disease, we do not know whether a specific autoantigen initiated the disease, nor do we know the time point at which the antigenic challenge occurred. As clinicians we are faced with late-term manifestations of the immunologic events that lead to clinical disease. Finally, in limited forms of cutaneous autoimmune disease, it is important to recognize distinct subsets with a favorable prognosis, for which less aggressive therapeutic modalities may be used. PMID- 11037618 TI - [Concepts for immune intervention in psoriasis vulgaris]. AB - Psoriasis vulgaris is a chronic inflammatory skin condition with a genetic background. Activated T-cells and their secreted products seem to play an essential role in the induction as well as the promotion of the psoriatic plaque. We will focus on some recent concepts on the immunopathogenesis of psoriasis highlighting the role of dendritic cells as initiators of the disease as well as the recruitment of disease specific T-cells. Concepts for immunointervention will be introduced. PMID- 11037619 TI - [Immuno-intervention in cutaneous lymphomas]. AB - Cutaneous lymphomas are a heterogeneous group of lymphoproliferative disorders. Most of the cutaneous lymphomas present a rather good prognosis because they disseminate to internal organs late in the disease process, if at all. Since radiotherapy and intensive chemotherapy have limited efficacy, immunological interventions are regularly used in this disease group. These interventions include unspecific treatment modalities such as Interferon-alpha, Interleukin-2, but also rather specific interventions like photopheresis, fusion toxins and as a new development, antigen-loaded dendritic cells. A new humanized anti CD20 antibody can be used for patients with cutaneous B-cell lymphomas, expressing the B-cell antigen CD20 in most of the cases. We expect that new information on the immunobiology of these diseases will allow more specific immunological interventions, providing the basis for a better therapy with optimal tolerability for patients suffering from cutaneous lymphomas. PMID- 11037620 TI - [Methods for increasing the immunogenicity of vaccines]. AB - In the past years, enormous efforts have been undertaken to develop vaccine strategies against cancer. The aim is to have the immune system generate what are called killer cells that can specifically recognize the tumor. The surface of tumor cells contains MHC/HLA antigens which present short-chain peptides of tumor specific antigens. A large number of these oligopeptide antigens have been characterized in recent years. They are now available for use as tumor-specific vaccines. The problem is, however, that the immune response of producing T killer cells is very inefficient when these oligopeptide antigens are injected. As the physiological function of these killer cells virus-infected cells, a process associated with substantial tissue damage, the immune system has learned to use these killer cells with reticence over the course of evolution, in other words, when the life of the host is threatened. This does not happen until pathogens start to spread via lymphogenous or hematogenous pathways. And then it takes a certain amount of time after the invader is present for replication to take place. Since the oligopeptide antigens used as vaccines have a very short half life in the tissue, not enough of them get to the lymph nodes and stay there for enough time to efficiently induce an immune response. Using a mouse model, we were able to show that the efficiency of the vaccine can be increased a million fold by directly injecting the vaccine into a lymph node or the spleen which imitates lymphogenous or hematogenous spread. The efficiency of the "inactivated vaccine" can be enhanced even more by continuous administration of the vaccine over several days, simulating an especially dangerous virus replication. The evidence gathered in this mouse model was transferred to a clinical trial. The melanoma-specific inactivated vaccine is infused directly into a lymph node of tumor patients. The infusion is continued for several days. Booster vaccines are given every two weeks. PMID- 11037621 TI - Ecdysone receptors and their biological actions. PMID- 11037622 TI - Chromatin-remodeling complexes involved in gene activation by the glucocorticoid receptor. PMID- 11037623 TI - Mechanism of action of estrogens and selective estrogen receptor modulators. AB - Estrogen, one of several sex steroid hormones, mediates its actions through the estrogen receptor. The estrogen receptor (ER) has two subtypes, ER alpha and ER beta, each of which predominates in specific tissues and organs. Cofactor proteins interact with the ER to maximize ligand-dependent transactivation of target-gene promoters. The estrogen response element is the final step in estrogen-mediated gene regulation, and current research is focused on alternate response elements. The resulting biologic action can vary according to the specific type of ER, cofactor milieu, response element, and ligand. Selective estrogen receptor modulators (SERMs) exhibit tissue-specific estrogen agonist or antagonist activity. The SERM raloxifene, which binds to ER and targets a distinct DNA element, may distinguish agonist vs antagonist activity by ER subtype and has unique activity among other SERMs because of its molecular conformation. Phytoestrogens, a potential alternative to hormone replacement therapy and for cancer prevention, do not consistently mimic estrogen's activity. Different types of phytoestrogens have different potencies, and taking high-dose supplements after menopause may not emulate the apparent benefits of lifelong consumption of phytoestrogen-rich diets. In conclusion, the complexity of estrogen action--through different ER subtypes, with various cofactors, on alternate response element--is further enhanced by ligands with selective estrogen activity. Additional research is needed to elucidate these pathways and the resulting biological effects. PMID- 11037624 TI - The role of protein kinase C in the development of the complications of diabetes. AB - Diabetes mellitus produces a state of chronic hyperglycemia which in turn leads to the development of severe complications including retinopathy, nephropathy, neuropathy, and atherosclerosis. Many different mechanisms have been put forward to attempt to explain how glucose elevations can damage these various organ systems. Protein kinase C activation is one of the sequelae of hyperglycemia and is thought to play a role in the development of diabetic complications. There are multiple mechanisms for its activation in the diabetic state and multiple downstream effects attributable to that activation. The role of protein kinase C activation in the development of the above-mentioned complications of diabetes is discussed in this chapter. In addition, the potential use of isoform-specific inhibitors of protein kinase C for the treatment of diabetic complications is proposed. PMID- 11037625 TI - Transcriptional regulation of the LH beta gene by gonadotropin-releasing hormone and the protein kinase C system. PMID- 11037626 TI - Angiotensin II and calcium channels. AB - Sixty years after its initial discovery, the octapeptide hormone angiotensin II (AngII) has proved to play numerous physiological roles that reach far beyond its initial description as a hypertensive factor. In spite of the host of target tissues that have been identified, only two major receptor subtypes, AT1 and AT2, are currently fully identified. The specificity of the effects of AngII relies upon numerous and complex intracellular signaling pathways that often mobilize calcium ions from intracellular stores or from the extracellular medium. Various types of calcium channels (store- or voltage-operated channels) endowed with distinct functional properties play a crucial role in these processes. The activity of these channels can be modulated by AngII in a positive and/or negative fashion, depending on the cell type under observation. This chapter reviews the main characteristics of AngII receptor subtypes and of the various calcium channels as well as the involvement of the multiple signal transduction mechanisms triggered by the hormone in the cell-specific modulation of the activity of these channels. PMID- 11037627 TI - Liposome targeting to tumors using vitamin and growth factor receptors. AB - Liposome-encapsulated anticancer drugs reveal their potential for increased therapeutic efficacy and decreased nonspecific toxicities due to their ability to enhance the delivery of chemotherapeutic agents to solid tumors. Advances in liposome technology have resulted in the development of ligand-targeted liposomes capable of selectively increasing the efficacy of carried agents against receptor bearing tumor cells. Receptors for vitamins and growth factors have become attractive targets for ligand-directed liposomal therapies due to their high expression levels on various forms of cancer and their ability to internalize after binding to the liposomes conjugated to receptors' natural ligands (vitamins) or synthetic agonists (receptor-specific antibodies and synthetic peptides). This chapter summarizes various strategies and advances in targeting liposomes to vitamin and growth factor receptors in vitro and in vivo with special emphasis on two extensively studied liposome-targeting systems utilizing folate receptor and HER2/neu growth factor receptor. PMID- 11037628 TI - Vitamins and homocysteine metabolism. PMID- 11037629 TI - Cobalamins and folates as seen through inborn errors of metabolism: a review and perspective. PMID- 11037630 TI - Advanced non-small cell lung cancer in the elderly: effective and well tolerated weekly gemcitabine and cisplatin regimen. A pilot study. AB - BACKGROUND: The frequency of advanced non-small cell lung cancer (NSCLC) increases with age and more effective and less toxic chemotherapy schedules are needed in elderly patients. Cisplatin-based regimens are considered the best treatment for advanced NSCLC, although they produce only a modest advantage in overall survival with considerable toxicity. METHODS: In the present study the activity and toxicity of a weekly gemcitabine and cisplatin schedule was evaluated in a small group of advanced NSCLC patients aged 68 years or more. Treatment consisted of gemcitabine 1000 mg/m2 i.v. and cisplatin 35 mg/m2 i.v., both given weekly on days 1, 8, 15 followed by 1 week of rest. RESULTS: Fifteen previously untreated patients entered the study; their median age was 72 years (range 68-76). One hundred and sixteen weekly administrations were delivered. The median dose-intensity was 614.5 mg/m2 per week for gemcitabine (82%) and 21 mg/m2 per week for cisplatin (80%). All the 15 patients were evaluable for response and toxicity. The overall response rate was 40% [95% CI = 16-68%]. The main toxicity was WHO grade III-IV thrombocytopenia that was recorded in 6 patients (40%). Other major toxicities were very low and no treatment-related deaths were reported. CONCLUSIONS: This schedule appears to be active, to have a favourable toxicity profile and can be considered in advanced NSCLC elderly patients. Of interest, the patients enrolled received high dose intensities of both drugs. PMID- 11037631 TI - [Biochemical markers of bone turnover and YKL 40 in ankylosing spondylitis. Relation to disease activity]. AB - BACKGROUND: YKL-40 is a glycoprotein produced by chondrocytes and synovial cells. The plasmatic levels of this metabolite increase in some pathologies such as rheumatoid arthritis and osteoarthrosis, so much so that it can be considered an effective marker of disease activity and in the therapeutic monitoring of these diseases. It has been interesting to dose a group of both male and female subjects affected by seronegative spondylarthritis, comparing this parameter with the disease activity indexes and with the bone turnover markers. METHODS: The study has been carried out on 48 subjects (26 males and 22 females) between 17 and 68 years affected by spondylarthritis, diagnosed in conformity with ARA standards. None of the patients carried out basic treatment or by glycocorticoids, and 22 patients took FANS when required. In these subjects the disease activity markers (VES, PCR, fibrinogen, mucoprotein) and some of the classic bone remodelling markers (blood calcium and phosphates, calciuria, phosphaturia, Ca++, Ntx, osteocalcine, bone isoenzyme of alkaline phosphatase, hydroxyproline, procollagen and YKL-40) were dosed. RESULTS: The comparison between different parameters pointed out that the highest values are obtained in subjects of most advanced age with the highest phlogosis indexes, without any correlation with sex. The quite interesting comparison shows a correlation between the bone remodelling indexes and YKL-40, being particularly remarkable when the disease is more aggressive or during relapse. CONCLUSIONS: It is then possible to confirm that, though preliminary, these data may suggest evaluations on wider case histories to research YKL-40 as a surgical monitoring marker in seronegative poliarthritis. PMID- 11037632 TI - [Histiocytic necrotizing lymphadenitis or Kikuchi-Fujimoto disease. Report of a case]. AB - Histiocytic Necrotising Lymphadenitis by Kikuchi-Fujimoto (NHL) is a rare disease of unknown etiopathogenesis, characterized by cervical lymphadenomegaly, fever and asthenia. It has a good prognosis with a complete functional recovery of the affected lymph nodes. In 1998 a 28 year-old patient (A. G.) was admitted in the Department of Internal Medicine, Garibaldi Hospital, Catania for fever, asthenia and cervical lymphadenopathy. Hemato-chemical tests performed during hospitalization showed only relative leukopenia and a significant increase of ESR. An initial cervical lymph node biopsy made the diagnosis of Hodgkin's lymphoma. However, the favourable clinical course and the normalization of the previous altered hematochemical tests, induced to perform a second lymph node biopsy which led to diagnosis of NHL. The patient was given antibiotics and anti inflammatory drugs for ten days with a complete healing which persisted at a twelve-month follow-up. The conclusion is drawn that NHL of unknown etiopathogenesis and with a good prognosis is more frequent than what revealed by the data in the literature because of its insidious and aspecific clinical presentation. PMID- 11037633 TI - [Bilateral sclerosing lipogranuloma of the gluteal region with calcification. Report of a clinical case and review of the literature]. AB - A rare case of large bilateral sclerosing lipogranuloma with multiple calcifications of gluteal region is described in an old female patient affected by a cerebrovascular disease. The lesions appeared as firm, nontender, plaques, 9 10 cm in diameter, covered with hyperpigmented skin. This uncommon disorder is discussed on the basis of data obtained from an extensive literature review. The term "sclerosing lipogranuloma" was coined in 1950, and it defines a disease of the subcutaneous fat, which for a trauma or unknown reasons undergoes necrosis of fat cells with the release of fat droplets into intercellular spaces and a peculiar local sclerosing granulomatous reaction of fatty tissue. The cytosteatonecrosis and sclerosing lipogranuloma, post-traumatic or secondary to injection of exogenous oily substances, usually localized in the breast of women and in genitalia of men, are relatively well known. Sclerosing lipogranulomatosis of the orbita and eyelides, an infrequent but severe complication after endonasal surgery, has also been reported. Rarely, the lipogranuloma can be spontaneous or idiopathic or primitive. A particular form of genetic diffuse lipogranulomatosis is the Farber's syndrome, firstly described on 1947. In our patient, the absence of trauma seems to indicate a primitive lipogranuloma. The presence of an acute rheumatic syndrome responsive to corticosteroids, and the positivity of antimitochondrial autoantibodies are in accordance with the report of sclerosing multiple lipogranulomatosis associated with a lupus-like syndrome. Because of the long duration and the absence of acute local symptoms, this syndrome can be considered benign with favorable prognosis, but the physician should know it. PMID- 11037634 TI - [Juvenile primary refractory anemia with excess of blasts. Report of a case]. AB - Myelodysplastic syndromes (MDS) are diseases typical of the adult age, characterized by a clonal alteration of the totipotent staminal cell which causes an inefficient hemopoiesis, reduction of bone marrow cell number, increased bone marrow cell destruction, dysplasia of at least two of the three hemopoietic cell lines and by the tendency to evolve towards acute myeloid leukemia (AML). In patients with MSD, particularly in the advanced primary form, single or multiple chromosomal abnormalities can be found frequently, which may show up and/or modify themselves in any moment of the disease (multistep pathogenetic hypothesis) and whose severity influences significantly the prognosis of MSD patients. In November 1998, a 22 year old female patient (C. C.) was admitted to the Department of Internal medicine of the Garibaldi Hospital, Catania for anemia of unknown origin. The normalities of the hematochemical tests and of other instrumental examinations, induces to perform an osteo-medullary biopsy which revealed the presence of a typical MSD, refractory anemia with excess of blasts type. This disease in its primary form is rare in youth and has a high tendency to evolve in AML. The good clinical conditions of the patient, the absence of chromosomal abnormalities, the normal levels of HbF, the short time interval of the initial diagnosis induced to proceed to bone marrow transplantation, that, as shown by the data reported in the international literature, may give her a good quoad vitam prognosis. PMID- 11037635 TI - [Diabetes mellitus and spontaneous hypoglycemia. Report of a clinical case with extrainsular neoplasm]. AB - The authors report a case of hypoglycemia in a subject with NIDDM and CAD. The clinical syndrome, which was diagnosed as an induced form for a long time, was found to be spontaneous and caused by metastasised kidney neoplasm. This led to considerable problems of differentiated diagnosis owing to the concomitance of diabetes and vascular pathology. An adequate assessment of the clinical findings and the instrumental and biohumoral tests would have enabled a more rapid diagnosis, thus allowing a correct therapeutic approach to be adopted. PMID- 11037636 TI - The Heart Surgery Forum is now in Index Medicus/MEDLINE. PMID- 11037637 TI - [Digitized analysis of hand movements in psychiatry. Methods, clinical findings and perspectives]. AB - Numerous morbo- and pharmacogenic motor disorders are signs of psychiatric disease. In past, their exact and quantitative registration has not been possible or only possible using quite extensive procedures. Now we can use newer methods like the computer-aided analysis of hand movements, an elegant procedure allowing the detailed representation of discrete motor dysfunction. While these procedures are used with increasing tendency in neurology, their use aiming the investigation of psychiatric hypotheses is still at the beginning. Relevant application fields of the computer-aided analysis of hand movement are the differentiated and quantitative registration of movement disorders that could only be detected by use of rating scales so far; the registration of subclinical motor abnormalities leading to the detection of the participation of the motor system in certain subgroups of psychiatric patients; the differentiation of morbo and pharmacogenic motor disorders (e.g. hypokinesia induced by neuroleptics versus schizophrenic negative symptomatology); the registration of neurological soft signs (e.g. in schizophrenic patients); the individual dose adjustment accounting for the threshold of extrapyramidal motor side effects (EPMS) under neuroleptic treatment and the detection and documentation of clinical and subclinical tremor. For some of these application fields there already exist promising findings. Some newer results will be presented. Schizophrenic patients were found to manifest impairments of velocity and degree of automatization and a reduced regularity of repetitive hand movements (fine motor dysdiadochokinesia)- a possible indicator for a disturbance of cerebral development. Patients with Alzheimer's disease showed loss of automatization and well preserved peak velocity. This finding could be relevant for the early detection of Alzheimer's dementia. Distal EPMS induced by neuroleptics can be exactly documented and their course can be represented using this method. The quick and economical utilization of digitized analysis of hand movements makes this procedure suitable for a broader application in the ambulatory setting. PMID- 11037638 TI - [Mechanisms of cerebral autoregulation, assessment and interpretation by means of transcranial doppler sonography]. AB - Cerebrovascular autoregulation assures constancy of cerebral perfusion despite blood pressure changes, as long as mean blood pressure remains in a range between 50-170 mmHg. Static and dynamic myogenic mechanisms dampen sudden blood pressure changes. Neurogenic influences of sympathetic, noradrenergic fibers modulate primarily proximal, large diameter segments of cerebral arteries, but also small 15-20 microns diameter vessels. Parasympathetic, vasodilating impulses are of less influence. Monoaminergic brainstem centers such as the dorsal raphe nucleus, locus coeruleus or nucleus reticularis pontis oralis also influence vessel tone. Metabolic, local parenchymal and endothelial substances have major impact on cerebral vessel tone. Particularly important are nitric oxide, calcitonin gene related peptide, substance P, endothelin, potassium channels and autocoids such as histamine, bradykinin, arachidonic acid, prostanoids, leucotrienes, free radicals or serotonin. The clinical examination of autoregulation is mostly based on brief blood pressure changes induced by drugs such as angiotensin, phenylephrine or sodium nitroprusside, or by challenge maneuvers. Frequently, blood pressure is challenged by a tilt-table maneuver, the "leg-cuff"-method according to Aaslid, or a Valsalva maneuver. The analysis of coherence and phase relation between spontaneous or metronomic breathing modulation of blood pressure and brain perfusion also assesses autoregulatory function. Cerebral blood flow is determined by means of transcranial Doppler sonography, mostly of the proximal segment of the mid-cerebral artery. There is some controversy whether a decrease of cerebral blood flow velocity measured at this segment indicates vasodilatation at the insonated segment or reflects blood flow reduction due to decreased perfusion of down-stream vessel segments. Various clinical and animal studies are presented demonstrating diameter constancy of the insonated mid-cerebral artery segment and thus indicating that slowing of mid cerebral artery blood flow velocity as assessed by transcranial Doppler sonography is due to a decrease of down-stream perfusion. Direct, intraoperative measurements of vessel diameter confirm this conclusion. PMID- 11037639 TI - [Development and validation of a test for early diagnosis of dementia with differentiation from depression (TFDD)]. AB - Psychometric tests used for the early detection of dementia often are seen as too difficult or too complex. Classical neuropsychologic tests were not developed for this purpose. Sensitivity and specificity to discriminate "healthy" vs. "ill" are low. For measuring both dementive and depressive symptoms, so far no test has been published. The objective of this study was to develop a sensitive and specific test for dementia that is easy to administer and to evaluate. Moreover, it should discriminate dementia from depressive pseudodementia. With respect to former studies, items were selected that recognized patients in the beginning of the disease. Additionally, depressive symptoms were rated. With the items for dementia, 88 patients with dementia of the Alzheimer type, 52 patients with depressive disorder and 37 healthy elderly controls were investigated. In this group of already diagnosed patients, the test reached a sensitivity and specificity of 100 percent (healthy elderly controls vs. patients with Alzheimer's disease: n = 125, U = 0, p < 0.001; patients with depressive disorder vs. patients with Alzheimer's disease: n = 140, U = 0, p < 0.001; healthy elderly controls vs. patients with depressive disorder: n = 89, U = 485.5, p < 0.001). For the dementia items, the inter-rater-reliability was rs = 0.996 (p < 0.001, n = 18), for the depression items it was rs = 0.753 (n = 18, p < 0.001). The test retest-reliability was rs = 0.868 (p < 0.001, n = 35) for the dementia items and rs = 0.7 (n = 8, p < 0.05) for the depression items. These validation data will make the test useful for practitioners. Its ability to discriminate patients suffering from dementia of the Alzheimer type from healthy controls is comparable to tests consuming more time. PMID- 11037640 TI - [Early intervention following trauma. An overview of programs and their effectiveness]. AB - A powerful motivation for clinicians working in the field of trauma is the wish to prevent or minimize chronic posttraumatic reactions by early interventions. This paper illustrates the process and the elements of important programs that have been introduced into the psychological or psychiatric literature. Studies that have been done to evaluate these programs are discussed. With regard to early interventions given to traumatized victims regardless of whether they suffer from posttraumatic stress or not, it is clearly shown that the evidence that these programs have a substantial positive benefit is disappointingly scarce. However, with regard to the treatment of acute stress disorder, a brief cognitive-behavioral program appears to effectively prevent the development of chronic posttraumatic stress disorders. In summary, it has to be noted that much more research on this issue is needed. PMID- 11037641 TI - In vitro dissolution of cholesterol and brown pigmented gallstones: a comparison of MTBE, DMSO and BA-EDTA. AB - BACKGROUND: Gallstones are a common problem in Taiwan and surgical removal remains the essential treatment. Successful dissolution of the stones with chemical solutions and then removal by endoscopic or percutaneous methods have previously been reported. We designed this study to find the ideal agent for dissolving gallstones. METHODS: Twelve chemical solutions with dimethylsulfoxide (DMSO), methyl tert-butyl ether (MTBE) and ethylenediamine tetra-acetic acid (EDTA) in different mixtures were tested to investigate their ability to dissolve gallstones in vitro. The dissolution of stones was performed at 37 degrees C and each procedure was repeated five to seven times. RESULTS: The solvent containing DMSO/MTBE (1/1) had a higher dissolving capacity for cholesterol stones, with solubility reaching 96.8% after 6 hours. The solution containing DMSO/MTBE (7/3) had the maximal solubility for calcium bilirubinate stones, with solubility reaching 22.9% after 6 hours. Also, we found that the intact stones of calcium bilirubinate became fragmented after treatment with the DMSO/MTBE solution without stirring. CONCLUSIONS: The DMSO, MTBE and EDTA agents that we used effectively dissolved gallstones, especially cholesterol stones, in vitro. Further in vivo studies are necessary to confirm the efficacy and safety of these solvents before clinical application. PMID- 11037642 TI - Brain-damaged survivors after intrauterine death of a monochorionic twin. AB - BACKGROUND: Potential risks for a surviving twin after fetal death of a co-twin in twin-to-twin transfusion syndrome (TTTS) has been documented. Although some studies suggest ending a twin pregnancy after a single fetal death as soon as possible in order to minimize the risks of thromboembolic complications in the surviving twin, we are more concerned about the risks of a premature birth. In this study, we searched for a potential marker to predict thromboembolic complications in the surviving twin. METHODS: From 1993 to 1998, nine women in two teaching hospitals had pregnancies complicated by TTTS and the death of one fetus. In addition to routine ultrasound examinations and obstetric monitoring, all patients had disseminated intravascular coagulation tests. The outcome of the surviving fetus was recorded. RESULTS: Three patients had silent and minimal coagulopathy (33%) that revealed only the presence of D-dimer. Among these three patients, two had a disappearance of serum D-dimer, but the other one had persistent D-dimer levels for more than 5 weeks and delivered a fetus with a cerebral infarction. Except for the above-mentioned fetus, all surviving fetuses were normal and healthy and were delivered at the median gestational age of 33 (range, 31-36) weeks. Of the nine surviving children, the mean interval between fetal death of one twin and delivery of the healthy twin was 22.7 days, ranging from 3 days to 47 days. CONCLUSIONS: All patients having TTTS associated with one fetal death should be carefully monitored for coagulopathy; the appropriate time for delivery might depend on the duration of persistent D-dimer in the maternal blood. The long-term presence of D-dimer in the maternal serum may indicate a severe underlying thromboembolic complication in the surviving twin after intrauterine death of a monochorionic twin. Due to the increased risks of morbidity in premature births, the benefit of early termination of pregnancy in order to protect against subsequent sequelae in the surviving twin is minor and remains an area for future research. PMID- 11037643 TI - Perinatal management of congenital complete heart block. AB - BACKGROUND: The perinatal management of congenital complete heart block (CCHB) remains controversial. The purpose of this study was to present a therapeutic modality for CCHB. METHODS: We collected retrospective cases of all pregnant women admitted to our hospital between January 1992 and June 1999 whose babies developed CCHB antenatally. After a series of examinations, maternal, fetal and neonatal data were analyzed. RESULTS: Nine fetuses from six mothers (cases 1-6) in nine different pregnancies were studied. In case 1, both consecutive fetuses had CCHB and in case 2, all three consecutive fetuses had CCHB. The other mothers (cases 3-6) had only one fetus each with CCHB. Of the seven fetuses with isolated CCHB, four underwent observation only due to late-onset, or nonimmunologic CCHB, two received dexamethasone and/or intravenous immunoglobulin therapy because of the presence of hydropic signs, and one received dexamethasone at 23 weeks' gestation due to early-onset CCHB. Shortening fractions of the right ventricle had good compensation in four fetuses, without any treatment, and improving compensation in two of three fetuses receiving dexamethasone therapy. All seven fetuses were delivered smoothly and pacemakers were implanted shortly after birth. Two other fetuses had a poor outcome due to associated ventricular septal defect or hemoglobin Bart's disease. Furthermore, we gave dexamethasone (2 mg/day) instead of prednisolone (10 mg/day) for the next pregnancies of patients 3 to 5, beginning at 12 weeks of gestation. No fetal CCHB developed again. CONCLUSIONS: For pregnant women with previous fetal immunologic CCHB, early initiation of dexamethasone instead of prednisolone might be effective to cross the placenta and avoid recurrences. Dexamethasone is also effective for fetal CCHB of early onset, fetal hydrops or heart failure. Observation only is suggested for nonimmunologic CCHB and remote or late-onset immunologic CCHB. Other modalities were tried for very sick fetuses, but their effectiveness was not predictable. PMID- 11037644 TI - Vaginal fluid creatinine, human chorionic gonadotropin and alpha-fetoprotein levels for detecting premature rupture of membranes. AB - BACKGROUND: Diagnosis of premature rupture of membranes (PROM) is difficult in equivocal cases. The concentrations of human chorionic gonadotropin (hCG), alpha fetoprotein (AFP) and creatinine are high in amniotic fluid. The purpose of this study was to determine the usefulness of vaginal fluid hCG, AFP and creatinine measurements in the detection of PROM. METHODS: About 3 ml of normal saline was used to irrigate the posterior vaginal fornix and was collected for the measurement of hCG, AFP and creatinine. The control group included 10 normal pregnant women in the third trimester (> 28 weeks of gestational age). Levels of hCG, AFP and creatinine were compared with those of 10 women with confirmed PROM. RESULTS: The median levels of vaginal fluid hCG of normal pregnant women and pregnant women with confirmed PROM were 35.0 mIU/ml and 478.0 mIU/ml (p = 0.0046), respectively. For AFP, the corresponding values were 0.80 ng/ml and 54.24 ng/ml (p < 0.0001), respectively, and for creatinine, the values were 0.05 mg/dl and 0.95 mg/dl (p < 0.0001), respectively. All three markers were significantly higher in the experimental group than in the control group. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy for hCG were 80%, 70%, 72.7%, 77.8% and 75%, respectively. For AFP, these values were 90%, 100%, 100%, 90.9%, and 95.0%, respectively, and for creatinine, they were 90%, 100%, 100%, 90.9%, and 95%, respectively. The diagnostic value of AFP or creatinine level in vaginal washing may be better than that of hCG, though the difference was not statistically significant, probably due to the limited case number. CONCLUSIONS: Creatinine in vaginal fluid washings is a useful marker for PROM. It was less expensive and easier to measure than hCG and AFP, and appears to be more accurate than hCG. PMID- 11037645 TI - Does Ser364Pro mutation of connexin 43 exist in Taiwanese patients with Ivemark syndrome? AB - BACKGROUND: A previous study by Britz-Cunningham et al (N Engl J Med, 1995) indicated that a mutation of the connexin 43 (CX43) gap junction gene might be responsible for Ivemark syndrome. Ser364Pro substitution (TCA-->CCA) is the most common mutation located in the cytoplasmic tail domain of CX43. This domain may be an important part of the conductance channel of the gap junction. It may, therefore, result in heart anomalies and situs inversus during embryonic development, resulting in Ivemark syndrome. METHODS: We investigated 10 patients with Ivemark syndrome, 10 healthy individuals, one patient with Kartagener syndrome and one with polysplenia and situs inversus but without heart anomaly for this mutation. Seminested polymerase chain reaction (PCR) was performed using a DNA template from DNA extracted from peripheral blood cells. Direct sequencing was done after purification of the second round of PCR products. Then, the sequence was compared with the last 402 bp of the cDNA-coding region of CX43. RESULTS: No base changes were found in the patients with Ivemark syndrome or other patient groups at the previously reported CX43 residues of Thr326, Gln352, Ser364, Ser365 and Ser373. CONCLUSIONS: The results indicate that Ser364Pro mutation of CX43 did not exist in the 10 Taiwanese patients with Ivemark syndrome. Other genes responsible for the Ivemark syndrome should be further investigated. PMID- 11037646 TI - Intratracheal oxygen administration during bronchoscopy in newborns: comparison between two different weight groups of infants. AB - BACKGROUND: The development of ultrathin fiberoptic bronchoscopy (FB) has made the examination of neonatal airways a practical possibility. The aim of this study was to assess the effects of intratracheal oxygen (ITO) administration on blood oxygenation and carbon dioxide (CO2) changes during FB in different body weight infants. METHODS: Newborns suspected of having airway problems, but in a stable cardiopulmonary condition were studied. An ultrathin (outside diameter, 2.2 mm) fiberoptic bronchoscope that was modified by adding an external tube (internal diameter, 0.3 mm; outside diameter, 0.64 mm) to deliver oxygen was used. For ITO administration, a low oxygen flow rate of 0.1 l/kg/min was delivered directly into the trachea. Oxygenation and CO2 measurements were obtained at five different stages: 1) just before FB (baseline); 2) with the tip of the bronchoscope at the supralarynx; 3) with the tip at the carina without ITO; 4) with the tip at the carina with ITO; and 5) 15 minutes after FB. Forty infants were studied completely and divided into two groups according to their body weight: 1) the light-weight group (< 2,500 g), 21 infants; and 2) the heavy weight group (> or = 2,500 g), 19 infants. RESULTS: In both groups, arterial blood oxyhemoglobin saturation and oxygen tension decreased significantly (p < 0.05) when the tip of the bronchoscope advanced from the nostril to the supralarynx, and further decreased (p < 0.01) when at the carina level. Small infants had greater decrements of both oxygenation measurements (p < 0.05) than the large infants. After ITO administration, both oxygenation measurements increased significantly (p < 0.001) and returned to baseline following FB. Both end tidal pressure of CO2 (P(ET)CO2) and arterial CO2 tension (PaCO2) significantly increased from the baseline when the FB tip was advanced from the supralarynx to the carina (p < 0.05). During ITO administration, the PaCO2 increased (p < 0.01) but the P(ET)CO2 decreased (p < 0.001). After FB, both CO2 measurements returned to baseline. The pH only decreased during ITO administration. CONCLUSIONS: We conclude that FB causes significant hypoxemia and hypercapnia in newborns, especially in underweight infants. Appropriate ITO can be considered a safe and beneficial technique for maintaining oxygenation during FB. P(ET)CO2 monitoring may mask true blood CO2 retention during ITO administration. PMID- 11037647 TI - Resolution of refractory hepatic hydrothorax after chemical pleurodesis with minocycline. AB - Management of refractory hepatic hydrothorax is a challenge to physicians in clinical practice. We reported two patients with hepatic hydrothorax, non alcoholic cirrhosis and rapidly recurring pleural effusion. They failed to improve with diuretics and repeated thoracentesis. Refractory hepatic hydrothorax was successfully treated by minocycline-induced pleural symphysis. After pleurodesis, ventilatory function returned to normal in both patients. No recurrence of pleural effusion was noted. We suggest that minocycline pleurodesis is an alternative treatment for refractory hepatic hydrothorax because it is simple, safe and effective. PMID- 11037648 TI - Hyperamylasemia associated with endometroid carcinoma of the ovary. AB - Hyperamylasemia and alternations of serum isoamylases have been recorded in lung tumors, tubal disorders such as acute salpingitis and ruptured ectopic pregnancies and a variety of ovarian tumors, and they have been suggested as potential tumor markers. Hyperamylasemia was noted in a patient with a stage IIIC endometroid adenocarcinoma of the ovary. Serum levels of amylase decreased rapidly after removal of the ovarian tumor. In patients presenting with acute abdominal pain and elevated amylase levels, ovarian cancer should be considered in addition to acute pancreatitis. PMID- 11037649 TI - Endometrial stromal sarcoma of the vagina. AB - Endometrial stromal sarcoma is a rare tumor and has unique histopathologic features. Most tumors of this kind occur in the uterus; thus, the vagina is an extremely rare site. A 34-year-old woman presented with endometrial stromal sarcoma arising in the vagina. No correlative endometriosis was found. Because of the uncommon location, this tumor was differentiated from other more common neoplasms of the vagina, particularly embryonal rhabdomyosarcoma and other smooth muscle tumors. Although the pathogenesis of endometrial stromal tumors remains controversial, the most common theory of its origin is heterotopic Mullerian tissue such as endometriosis tissue. Primitive cells of the pelvis and retroperitoneum are an alternative possible origin for the tumor if endometriosis is not present. According to the literature, the tumor has a fairly good prognosis compared with other vaginal sarcomas. Surgery combined with adjuvant radiotherapy appears to be an adequate treatment. PMID- 11037650 TI - [Randomized biopsies (R.B.) in initial superficial bladder tumors (I.S.B.T.). Why?]. PMID- 11037651 TI - [Pyelocalicial diverticula]. AB - OBJECTIVE: To review the embryological and clinical aspects of the different types of pyelocaliceal diverticula, with special reference to the differential diagnosis and treatment. METHODS: Images of type I and II pyelocaliceal diverticula are shown. The conditions that cause difficulty in making the differential diagnosis are discussed. RESULTS: Urography continues to be the diagnostic method preferred and is sometimes aided by retrograde ureteropyelography. CONCLUSIONS: Pyelocaliceal diverticula are cystic eventrations of the upper urinary tract lying within the renal parenchyma that communicate through a narrow channel into the main collecting system. They occur in 0.2 to 0.5% of the population and are congenital in origin. Calyceal diverticula are frequently found incidentally on routine excretory urograms, but patients may complain of flank pain, hematuria or recurrent urinary infections. In the past, treatment required open renal surgery. Endourologic procedures are widely utilized today. PMID- 11037652 TI - [Brachytherapy of the prostate]. AB - OBJECTIVE: To present our results with brachytherapy in the treatment of localized cancer of the prostate. METHODS: From September 1999 to April 2000, 20 patients with T1 and T2 cancer of the prostate were treated with brachytherapy. The method utilized is described and the results are presented. The advantages of iodine-125 implantation using transrectal ultrasound are discussed. RESULTS: The morbidity was low and were mild irritative symptoms of the lower urinary tract that were managed without difficulty with medical treatment. Determination of PSA three months after the procedure showed good results in 80% of the cases. CONCLUSIONS: There is an increasing number of cases diagnosed with localized cancer of the prostate. Good results have been achieved with radical surgery and radiotherapy. Although the number of patients with 10 years follow-up is small, brachytherapy of the prostate has achieved good results with a low morbidity. This therapeutic modality should be offered to the patient with localized cancer of the prostate and the patient allowed to decide on the type of treatment. PMID- 11037653 TI - [Loss of heterozygosity in the 9p21 region as an inactivation mechanism of the p16 suppressor gene]. AB - OBJECTIVE: To determine the loss of heterozygosity (LOH) on 9p21 (locus D9S1747) in patients with renal carcinoma by analysis of microsatellite polymorphisms. METHODS: 40 patients with sporadic renal cancer were studied. LOH on 9p21 was performed by analysis of microsatellite polymorphisms. RESULTS: 23.7% showed LOH on 9p21. No correlation was found between this genetic alteration and tumor features. CONCLUSIONS: LOH on 9p21 was found in 23.7% of the patients in this series. LOH was found in 26.9% of renal cell carcinomas, 25% of papillary carcinomas and 25% of Bellini duct carcinomas. LOH was not found in the other histological types. PMID- 11037654 TI - [Bellini's carcinoma. Our experience]. AB - OBJECTIVE: To present our experience with Bellini duct carcinoma, a rare form of renal adenocarcinoma with well-defined histological, cytogenetic and immunohistochemical characteristics. The literature is reviewed. METHODS/RESULTS: We reviewed the records of 430 patients with renal tumor that had been treated over a 10-year period. Only 6 cases with Bellini duct carcinoma were found. The mean age was 60 years, all patients were male and hematuria and lumbar pain were the most common clinical manifestations. Diagnosis was made by imaging techniques (US, urography, CT). The definitive diagnosis was based on the histological and immunohistochemical findings following nephrectomy. The one-year and three-year survival rates were 50% and 0%, respectively, indicating the aggressive nature of this variant of renal adenocarcinoma. CONCLUSIONS: Bellini duct carcinoma is an uncommon variant of renal carcinoma with nonspecific clinical features. It is a very aggressive tumor type whose treatment is only by nephrectomy and the outcome is poor. PMID- 11037655 TI - [Radical cystectomy in the treatment of cancer of the bladder]. AB - OBJECTIVE: To analyze retrospectively the efficacy of radical cystectomy alone in the treatment of transitional cell carcinoma of the bladder. METHODS: 125 patients who underwent radical cystectomy were evaluated. The mean follow-up was 62 months. At the time of the study, 65 patients were alive (3 with bladder tumor and 1 with a second primary) and 60 patients had died (50 from bladder cancer and 10 from other causes). Nine patients were lost to follow-up. The Kaplan-Meier method was used for the survival analysis and the log-rank test for the comparison of the variables. RESULTS: The overall survival at 5 years was 50% and the cancer-specific survival was 56%. By tumor stage, the cancer-specific survival at 3 and 5 years were respectively: 83% and 85% for pT1, 78% and 70% for pT2, 52% and 42% for pT3, 24% and 12% for pT4 and 14% for pN+ (p < 0.0001). No differences were found between stages pT2a (73% and 68%) and pT2b (71% and 53%) (p = 0.2). The survival was significantly higher in patients with no residual tumor in the cystectomy specimen (pT0) (93% and 83%) than in those with residual tumor (60% and 53%) (p = 0.03). CONCLUSIONS: Radical cystectomy alone in the treatment of transitional cell carcinoma of the bladder was found to be effective in patients with tumor stage pT2. It is less effective in patients with tumor in the advanced stages (pT3 or pT4) or lymph node invasion. Radical cystectomy is an overtreatment in patients with no residual tumor in the cystectomy specimen. PMID- 11037656 TI - [Versatility of Snodgrass technique for the correction of different types of hypospadias]. AB - OBJECTIVE: To present our experience with the Snodgrass technique for primary hypospadias repair and for reoperations. METHODS: From May 1996 to August 1998, the Snodgrass technique was performed on 97 patients with hypospadias; 72% were distal, 19.5% were midshaft and 8.5% were proximal. Of these patients, 8.5% had a previous hypospadias repair. The patients were followed for a period of two years. RESULTS/CONCLUSIONS: Complications, including fistulas, occurred in 4 patients. The cosmetic results were excellent and there were no complications in patients who had a previous urethroplasty. PMID- 11037657 TI - [Conservative surgery of renal carcinoma in a case of synchronic bilateral involvement, with large size tumors and different histological type]. AB - OBJECTIVE: To report a case of a large, bilateral, synchronous renal tumor. METHODS/RESULTS: Treatment by conservative surgery achieved good results. The histological analysis showed two different carcinomas. The most most important steps of the surgical procedure are described. Renal preservation in patients with malignant tumors is discussed. CONCLUSIONS: The case described herein is rare since the tumors were synchronous and of uncommon histological type (one of the tumors was a chromophobic carcinoma). Despite the large size of the tumors, they were successfully managed by conservative surgery. PMID- 11037658 TI - [Pacinian neurofibroma of the vulva]. AB - OBJECTIVE: To report on a rare case of Pacinian neurofibroma of the vulva. METHODS: A 27-year-old patient who presented with a vulvar tumor is described. Patient evaluation showed no other remarkable findings. RESULTS: The nodule was resected without difficulty. The histopathological analysis-demonstrated Pacinian neurofibroma with abundant concentric laminar structures. The immunohistochemical (CD34+), ultrastructural and histological analyses showed perineural cells. No signs of neurofibromatosis were found. CONCLUSIONS: Pacinian neurofibroma may present in the vulva and could probably arise from the so-called perineural fibroblasts. PMID- 11037659 TI - [Solitary pulmonary nodule in a patient with treated carcinoma of the renal pelvis: metastatic disease?]. AB - OBJECTIVE: To describe a case of a solitary pulmonary nodule in a patient that had been treated for carcinoma of the renal pelvis. METHODS: A solitary pulmonary nodule was detected on the chest film of a patient that had been treated for carcinoma of the renal pelvis. The characteristics of the nodule are described and its diagnosis, with special reference to metastasis and primary pulmonary carcinoma, is discussed. RESULTS: The anatomopathological study demonstrated a chondroid hamartoma. CONCLUSIONS: A metastatic or a primary tumor is suspected when a solitary pulmonary nodule is detected in a patient that has been previously treated for urothelial carcinoma. However, other types of lesions with a more favourable outcome cannot be discarded, such as chondroid hamartoma as in the case described herein. PMID- 11037660 TI - [Incomplete sagittal septum of the bladder]. AB - OBJECTIVE: To describe an additional case of incomplete sagittal septum of the bladder. METHODS: A case of incomplete sagittal septum of the bladder in a young, asymptomatic patient with paralysis of both lower limbs is presented and literature is briefly reviewed. RESULTS: The diagnosis was made by intravenous urography performed five years after the accident that had caused the paralysis. Radiological evaluation showed an "apple heart" bladder with reduced capacity. No treatment was required. CONCLUSIONS: Incomplete sagittal septum of the bladder is rare. To our knowledge, only 7 cases have been previously reported. Incomplete sagittal septum is like complete septum, but both cavities communicate anteriorly or distally, according to the direction and depth the septum protrudes into the bladder. PMID- 11037661 TI - [Basal cell carcinoma of the scrotum. A rare localization linked to a bad prognosis]. AB - OBJECTIVE: To present a case of basal cell carcinoma of the scrotum. METHODS: In a review of 56 scrotal tumors, we found 4 primary neoplasias, 3 benign mesenchymal tumors and one malignant tumor, the basal cell carcinoma of the scrotum described herein. RESULTS: A 52-year-old patient with no remarkable urological or dermatological history, complained of an excrescence in the right hemiscrotum that he had noted for several years. A clinical diagnosis of basal cell carcinoma of the scrotum was made and the lesion was surgically excised. Pathological analysis of the surgical specimen confirmed the diagnosis. CONCLUSIONS: Basal cell carcinoma is the most common type of skin cancer in the middle-aged and elderly, but localization to the scrotum is rare. It is a tumor that grows locally and rarely metastasizes, although scrotal tumors are much more aggressive and patients should therefore be followed very closely after resection of the tumor. PMID- 11037662 TI - [Well differentiated mucus-producing adenocarcinoma of the right renal pelvis. Presentation of a case]. AB - OBJECTIVE: To present a rare case of well-differentiated mucus-producing adenocarcinoma of the renal pelvis. METHODS: Patient history, the results of the complementary tests after surgery, treatment and anatomopathological findings are presented. RESULTS/CONCLUSIONS: Well-differentiated mucus-producing adenocarcinoma of the renal pelvis is rare and is often associated with lithiasis, a long history of obstruction and inflammation. Nests of tumor cells surrounded by abundant mucus is a characteristic histological finding in this tumor type. PMID- 11037663 TI - [Retroperitoneal ganglioneuroma in childhood]. AB - OBJECTIVE: An incidentally discovered retroperitoneal ganglioneuroma in a 4-year old child is presented. METHODS: Ultrasonography and CT were performed. The surgical specimen was analyzed by macroscopic, histological and immunohistochemical techniques. RESULTS: US and CT localized a retroperitoneal mass independent from the left kidney and adrenal gland. The histological study showed a fascicular proliferation with myxoid and fibrillar areas mixed with mature ganglion cells. These cells were positive for neurofilament and neuron specific enolase. The patient had a favorable outcome with no signs of tumor recurrence. CONCLUSIONS: Ganglioneuroma is a rare, completely mature tumor that has to be differentiated from neuroblastoma. PMID- 11037664 TI - [Epidermoid carcinoma of the penis with vertebral metastasis treated with decompression and anterior fixation]. AB - OBJECTIVE: To describe a case of epidermoid carcinoma of the penis with vertebral metastasis. METHODS/RESULTS: A patient with epidermoid carcinoma of the penis underwent penile resection. One year postoperatively the patient presented with paraparesia and lumbar pain, and was diagnosed as having vertebral metastasis at L3. Surgical resection of the tumor was performed with resection of L3 and arthrodesis of L2-L4 via the anterior retroperitoneal approach. The anatomopathological findings were compatible with metastatic epidermoid carcinoma. The rarity of bony metastases from this tumor and the clinical course of the patient are discussed. CONCLUSIONS: In patients with lumbar pain and a previous history of malignant disease, metastasis should be suspected even in those tumors that are considered to have behave well. PMID- 11037665 TI - Intralesional recombinant interferon alpha-2b in Peyronie's disease. AB - OBJECTIVE: To evaluate interferon alpha-2b (IFN) in the treatment of Peyronie's disease (PD) since IFN exerts antifibrotic action through collagen synthesis inhibition and fibrolysis stimulation. METHODS: The study comprised 34 patients, aged 31 to 63, with clinical and ultrasonographic (US) diagnosis of PD, who gave their consent to enter the study. They had the disease for 10.1 +/- 5.6 (2-22) months. Ten million IU of IFN were injected intralesionally, twice weekly for 14 weeks or less if there was complete remission. Clinical evaluation included penis angle at erection, sexual dysfunction (pain, possibility of intercourse) and palpable plaque. Plaque size was evaluated by US. Systemic and local adverse reactions, and anti-IFN antibodies were monitored as well. RESULTS: Sexual dysfunction disappeared in 19/24 (79.2%) patients with this disorder, palpable lesions in 21/34 (62%), angle at erection in 15/32 (47%), and pain in 16/17 (94%). Complete clinical response was achieved in 16/34 patients (47%). Ultrasonographic response rate was 88%, (53% complete). Plaque size decreased from 56.7 +/- 42.9 (median: 35.4) before treatment to 12.7 +/- 22.6 mm2 (median: 0) (p < 0.00001; Wilcoxon's paired test). Clinical and US responses correlated. No patient showed progression. Eight of 9 patients in whom other treatments had failed responded to IFN therapy (5 complete). The main systemic adverse reaction in most patients (mild or moderate) was the flu-like syndrome expected for IFN. Local reactions, more related to the administration procedure than to IFN itself, were small hematoma (10 patients), edema (3), cysts that were excised surgically (2), and venous leak (1). No patient developed anti-IFN antibodies. CONCLUSIONS: IFN treatment can be a suitable option for the management of PD. The results appear to be better than those achieved with other procedures. Further work should include comparative studies, long-term follow-up of treated patients, and alternative ways of administration. PMID- 11037666 TI - [Application of geographical information systems in epidemiological studies exemplified by the ISAAC study in Munich]. AB - Geographical Information Systems (GIS) are increasingly applied as modern tools for analysis and visualization of health-related spatial data, especially in epidemiological research. GIS are used by medical researchers and executives in the public health service. A community-based survey was conducted according to the phase II protocol of the International Study of Asthma and Allergies in Childhood (ISAAC) in Munich. The spatial patterns of disease incidence were analysed and related to exposure data by GIS. The prevalence study on fourth grade pupils (n = 3354) and school beginners (n = 2890) was conducted during the school term 1995/96 in Munich. Parental questionnaires and measurements of lung function and immunological parameters were used. The questionnaire data were integrated in a GIS database. In this paper we discuss methodological aspects of GIS-based spatial analysis related to epidemiological data. In addition, we investigate whether there were spatial clusters of children with wheeze in the last 12 months of a magnitude unlikely to occur by chance and which could indicate local health risks. The study was based on permutation tests where global and local methods were applied. No spatial clusters of children with asthma symptoms were identified in the city of Munich. PMID- 11037667 TI - ["Interface problems" in medical rehabilitation: development of a brief questionnaire for assessing the need for information and communication among general practitioners]. AB - Against the background of current and future demographic changes the relevance of medical rehabilitation in the German health care system is generally acknowledged. It is a central element of treatment programmes especially for elderly and chronically ill patients. In recent years, problems due to lack of co operation and communication as well as insufficient links between acute care and rehabilitation treatment have found increasing attention. Previous attempts to increase involvement especially of general practitioners in the rehabilitation process were only slightly successful. In a pilot study conducted by the Rehabilitationswissenschaftlicher Arbeitskreis Schleswig-Holstein a new procedure was tested to increase involvement of general practitioners. A brief questionnaire was attached to the report of discharge of rehabilitation patients and was sent to 130 practitioners. With this questionnaire the general attitude and need for information of the practitioners should be analysed. Knowledge about these variables should help to improve communication and co-operation. The response rate was quite high: about 75% of the contacted practitioners answered the questionnaire. All practitioners described medical rehabilitation as important and also had a quite positive attitude toward the same. Two-thirds of the physicians were interested in more intensive contacts and requested further information. PMID- 11037668 TI - [Therapy of cancers from the viewpoint of the Austrian population]. AB - The aim of this study was to survey the level of knowledge of the Austrian population about cancer treatment and to explore the ratings of the respondents concerning the importance of different modalities of treatment. A representative sample of Austrians > or = 15 years of age (n = 2073) was interviewed. Out of the respondents 94% claim that they have heard or read about chemotherapy. With regard to surgery and radiotherapy, this is stated by 83% and 73%, respectively. Hormonal therapy, immunotherapy and complementary therapy are known by a much smaller percentage (34% and less), whereas 52% claim that they have heard about homeopathy as a form of cancer treatment. The self-assessment of knowledge shows that 64% of all respondents state to have some knowledge about chemotherapy (surgery: 45%, radiotherapy: 26%). Persons with limited formal education report to have a lower level of knowledge. Surgery is rated to be the most important regimen, followed by chemotherapy and radiotherapy. Respondents with some knowledge about a certain treatment give higher ratings concerning the importance of this modality. The survey shows that a high percentage of the Austrian population has heard or read about chemotherapy and claims to have some knowledge about it. Especially the latter is not found for other treatments. Surgery, chemotherapy and radiotherapy are seen as the most important modalities of treatment. Individuals with a low level of education constitute an especially important target group for information about cancer treatment. PMID- 11037669 TI - [Inpatient treated, mono-symptomatic childhood enuresis--results of follow-up in adulthood]. AB - 32 formerly monosyndromatic enuretic patients who had been treated for this condition between 1980 and 1992 were compared with a matched control group from the general population in respect of sociobiography, psychopathological and dimensional factors (depressiveness, satisfaction with life, global functioning). Most of the former patients did not fulfil diagnostic criteria for an ICD-10 diagnosis at follow-up, although there was a 37% vs. 9% difference between former patients and controls in this respect--without a clear diagnostic pattern of such disorders. Furthermore, former patients had slightly higher depression scores and slightly lower global functioning than controls at follow-up. These results confirm that childhood enuresis has a low negative predictive value concerning the development of psychiatric disorders, although it may constitute a vulnerability factor. PMID- 11037670 TI - [Dental care in cross-cultural networks-- case management--approaches in group prevention]. AB - Oral epidemiology studies of previous years have shown an increasing difference in caries cases in respect of different social strata. Thus, frequency of caries cases is related to social status. High rates of caries prevalence are found especially among children from typical areas of welfare problems. Already, today every fifth child is born into a family of immigrants. In areas of typical social deprivation their share is about 40% and more. Since the previous educational campaigns for social fringe groups have hardly shown any positive effect on dental health, new strategies are necessary. In a community of Hanover with low socio-economic status and a generally high caries level, the treatment strategies of dental care for the young have centred on the case-management approaches of social welfare programmes since the early 90s. Beside the expanded basic preventive programme, which includes application of a fluoride varnish for children, social compensatory measures with intercultural networks are also being taken. This concept shows very clearly that the dental health of children living in areas of social disorganisation can be effectively improved by means of these strategies. For the future, dental care for these children requires more intercultural competence and more knowledge of social welfare work by adolescent dental care providers. PMID- 11037671 TI - [Psychosocial stress and utilization of medical services after coronary bypass operation]. AB - This study examines the relationship between psychosocial stress and social support before coronary surgery and the amount of health care utilization in a sample of 136 patients during postoperative hospitalization. The aim of the study is to test the hypothesis that there is a correlation between a high psychosocial stress profile and the utilization of medical care (so called high utilizers). The sample consists of 80.7% men and 19.3% women aged between 31 and 78 years (mean 64; sd 9.1). In this first data analysis psychosocial impact is assessed by anxiety, depression and social support (HADS-D, F-SOZU). Detailed somatic factors concerning severity of the illness, inpatient course and the utilization of health care (medication, technical examinations, consultations) are assessed by means of a documentation system. With regard to anxiety, depression and social support the sample is located within the normal range. In contrast to our expectations the results show that high scores of anxiety and depression as well as a low level of emotional support do not correlate significantly with an increased use of medication, the number of consultations and technical examinations. Furthermore no correlation has been found between the length of hospitalization and preoperative comorbidity as compared to the mentioned psychosocial stress variables. On the other hand the data analysis showed that about 30% of the patients during the postoperative period utilize about half of the total amount of the different medical treatments. In the postoperative period these high utilizers cannot be distinguished from the other patients, neither by sociodemographic variables nor by means of an increased psychosocial stress or severity of illness. PMID- 11037672 TI - [Indirect costs and time costs of (ambulatory) rehabilitation of mothers with psychosomatic disorders who have preschool children]. AB - Ambulatory rehabilitation concepts for women with psychosomatic disorders and with pre-school children are rare and moreover not yet assessed. An economic concept for the evaluation of indirect costs and (patient) time costs is being developed in this article and applied to an ongoing ambulatory rehabilitation programme for mothers at the Hanover Medical School. In health economic evaluations time cost is expressed by loss and reduction of working time, time for housework, and leisure time. These are indirect cost items (working time) and direct non-medical costs (housework and leisure time). To estimate the loss of working time (and hence production loss) the human capital approach and the frictional cost approach can be applied. Loss of time due to housework can be estimated either by the production of goods and services or by the opportunity costs of the equivalent working time. Loss of leisure time can be partial or total whereas a total loss and a loss of working time are considered to be analogous. The health economic evaluation of the ambulatory rehabilitation programme for mothers is designed as a randomised controlled study with repeated data collection. The parameters of indirect and direct non-medical costs are measured at the beginning of the rehabilitation programme and until twelve months later by means of questionnaires, face-to-face and telephone interviews. So far, results of the evaluation show that the actual time cost of the rehabilitation programme is DM 6,162 for each mother and the time cost because of the utilisation of the health care system is DM 996 per four weeks. Therefore, the patient costs are obviously higher than the direct medical costs for the programme which makes it clear that taking into account the costs of the patient (especially the time costs) can make a decisive difference in the evaluation of alternative treatment programmes and may possibly reverse the advantages of an alternative. PMID- 11037673 TI - [Psychosocial competence of general practitioners in managing psychiatric patients: initial results of a questionnaire study]. AB - OBJECTIVE: Given the frequency of patients with psychosocial problems and mental disorders in primary care our study focuses on the relationship between the psychosocial qualifications of general practitioners and their management of these patients. METHOD: A questionnaire was sent to all general practitioners in Hessen asking for sociodemographic and profession-related data as well as their psychosocial competence and their procedure during a regular day in practice. Based on 396 questionnaires we compared five groups of general practitioners with different psychosocial qualifications, proportionate incidence of diagnoses, medical procedures and referrals to specialists and hospitals. RESULTS: The average age of the participants of our study is 45 years, they have been working for 14 years. 163 general practitioners without and 59 with a special interest in psychosocial qualification and 174 psychosocially qualified physicians participated in the study. Of a total of 65 patients seen during the reference day 18 were diagnosed as suffering from mental disorders. Their number increased with greater psychosocial competence of the physicians. Psychosomatic and reactive disorders are the most frequent mental disorders to be seen in primary care with nearly 60%. Verbal therapy is often applied, procedures of the so called psychosomatic basic care and prescription of psychotropic drugs play only a minor role. Nearly every fourth patient is referred to other specialists, referrals for inpatient treatment occur with 1.3% only. DISCUSSION: Because of the low response rate our study is not representative. Nevertheless the results permit some conclusions concerning the relationship between psychosocial qualification of general practitioners and their management of patients with mental disorders. PMID- 11037674 TI - [Responsibilities and status of the medical expert]. AB - Expertising by assessment is an elementary, specific medical assignment, which in the history of medicine has early been linked to medical activity. No community can do without it, though all along critical voices have accompanied this occupation. The expert is in a field of tension between the claims of the individual and the reasonable interests of the community and the insurance companies. This results in points of attack and criticism challenging for the expert and his function. There should be consensus of opinion in respect of the responsibility and position of the medical expert among the members of the medical profession. This paper will make a contribution to this problem regarding the role of the expert and his self-image. PMID- 11037675 TI - [Social wealth in Germany II]. PMID- 11037676 TI - Production & characterization of monoclonal antibodies to Mycobacterium tuberculosis. AB - BACKGROUND & OBJECTIVES: Monoclonal antibodies (MAbs) against Mycobacterium tuberculosis H37Rv culture filtrate (CF) were raised by immunizing BALB/c mice and characterization was done. Attempts have been directed towards identifying mycobacterial antigens in biological fluids by employing polyclonal and monoclonal antibodies specific for M. tuberculosis. Immunohistologic studies, using MAbs for the localization of whole or fragmented bacilli in the biopsy specimens were also carried out. METHODS: Intrasplenic IS and intraperitoneal i.p. routes of immunization, were compared. The MAbs were characterized for their isotype, binding specificity, nature of binding epitope, reactivity in immunoassays etc. RESULTS: IS and i.p. routes of immunization, were compared and i.p. was found superior. Ten MAbs designated TRC 1-10 were produced. Of these, 7 MAbs, TRC 1-7 reacted with the 30/31 kDa doublet (antigen 85 complex), TRC 8 with 12 kDa in addition to 30/31 kDa and TRC 9 and 10 with the 24 and 12 kDa antigens respectively. Six MAbs were classified as broadly cross reactive and 2 showed limited cross reactivity. TRC 8 and 10 showed species specificity. Employing TRC 8 in sandwich ELISA, antigen was detected in sera from 17 of 25 pulmonary tuberculosis patients and 3 of 20 controls. TRC 8 was found to be useful in detecting antigens specifically in M. tuberculosis and M. leprae infected tissues, by immunoperoxidase staining. INTERPRETATION & CONCLUSION: TRC 8 was found to be restricted in its reactivity to M. tuberculosis complex and M. leprae. TRC 8 may prove useful in immuno-diagnosis of tuberculosis. PMID- 11037677 TI - Utility of polymerase chain reaction using two probes for rapid diagnosis of tubercular pleuritis in comparison to conventional methods. AB - We have used polymerase chain reaction (PCR) with IS6110 and a new set of primers from an insertion element like repetitive sequence, (TRC4) to detect Mycobacterium tuberculosis in pleural effusion samples from 50 patients having pleuritis. The results of PCR were compared with the results of conventional methods like smear, culture and adenosine deaminase activity. Thirty six specimens were positive and 14 were negative by PCR. Among the 36 samples, 33 were from patients with clinical evidence of tuberculosis including response to anti-tuberculosis therapy. Only six samples were positive by the gold standard which is culture, and three were positive by smear. The measurement of adenosine deaminase activity classified 19 samples as positives. The overall sensitivity and specificity of PCR was 100 and 85 per cent respectively. PCR using IS6110 and TRC4 primers is a sensitive test as compared to conventional tests for detection of M. tuberculosis from pleural fluid samples of patients with tubercular pleuritis. PMID- 11037678 TI - Leprosy affected beggars as a hidden source for transmission of leprosy. AB - BACKGROUND & OBJECTIVES: Despite the large scale implementation of multidrug therapy (MDT), the incidence rates of leprosy have not declined in several hyperendemic countries. Before searching for non-human reservoirs of leprosy it would be necessary to look for hidden human sources. This would include destitute leprosy affected persons who resort to begging and operate in congested areas. Hence this study was undertaken. METHODS: One major town and three semi-urban areas in Vellore district of Tamil Nadu and Chittoor town in Andhra Pradesh were purposefully selected for the study. All beggars in these towns were systematically identified and examined by allopathic doctors. Skin smears were examined for bacteriological index. RESULTS: Among the 193 beggars screened, 58 had leprosy. Of these 10 were smear positive. Several beggars, although living separately, were in touch with their relatives. Most beggars were pavement dwellers and regularly begged at places of worship, bus stands and shopping centres. INTERPRETATION & CONCLUSION: The fact that nearly 20 per cent of the leprosy affected beggars were skin smear positive highlights the need for regular screening and treatment of such beggars. Those positive should be actively treated and their close contacts frequently screened. This hidden reservoir should be completely eliminated. PMID- 11037679 TI - Changing scenario of cryptococcosis in a tertiary care hospital in north India. AB - BACKGROUND & OBJECTIVES: With the increase in the number of patients of AIDS, the incidence of cryptococcosis is on the rise in India. It was therefore considered important to evaluate the predisposing factors, laboratory investigations and outcome of patients with cryptococcosis in this changed scenario. METHODS: We assessed 58 patients with cryptococcosis retrospectively over a five year period (January 1995-December 1999) at the Nehru Hospital, Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh. RESULTS: The annual incidence of cryptococcosis in PGIMER, Chandigarh has increased about 15 fold from 1970-1982 (pre AIDS era) to 1995-1999 (present series). Of the 47 patients studied for predisposing factors, 36 patients were identified with predisposing factors, HIV infection (57.4%) was the commonest followed by haematologic malignancies (6.3%) and renal transplant (4.2%). Forty one patients were diagnosed by isolation of the organism as well as antigen detection in cerebrospinal fluid/serum, 9 by isolation alone and 8 by antigen detection alone. Quantitative antigen titres were done in 38 patients and a significantly higher (P < 0.01) antigen titre (> 512) was observed in HIV positive patients as compared to HIV negative patients. All isolates tested were of Cryptococcous neoformans var neoformans biotype and no resistance to antifungal agents was noted. Twenty of 41 patients receiving treatment improved. The results were compared with other studies available from India. INTERPRETATION & CONCLUSION: The incidence of cryptococcosis is on the rise in this part of north India and this can be attributed to an increase in AIDS cases. PMID- 11037680 TI - Transfusion transmitted diseases in haemophilics from western India. AB - BACKGROUND & OBJECTIVES: Transfusion related human immunodeficiency virus (HIV), hepatitis B virus (HBV) and hepatitis C virus (HCV) infections have been a major cause for morbidity and mortality in the haemophilic population in the west. The prevalence of these markers of transfusion transmitted viral diseases in severe and moderate haemophilia patients was studied. METHODS: The seropositivity for these viral markers was evaluated in 400 haemophilics (323 severe and 77 moderate) in a 5-year survey starting from 1995. First 188 of these patients were also tested for HCV. Serological tests for HIV, HBsAg and HCV were done by third generation ELISA; positive samples were also confirmed by Western blot. RESULTS: Fifteen of the 400 patients were found to be HIV positive (3.8%), 24/400 were HBsAg positive (6%) and 45/188 (23.9%) were positive for HCV (28 for both non structural and core antigen, 13 for core only and 4 for non-structural antigen only). The lowest age of HIV positivity was 12 yr and that of HCV positivity was 8 yr. INTERPRETATION & CONCLUSION: The above study shows a reduction in blood product related HIV transmission in severe and moderately affected haemophilics but more stringent policy for blood product usage, universal hepatitis C screening, hepatitis B vaccination and continuous awareness programmes for medical staff, general public and patients is needed to reduce the incidence of these diseases in haemophilics. PMID- 11037681 TI - Effect of basolateral amygdala & ventromedial hypothalamic lesions on ingestion & taste preference in rat. AB - BACKGROUND & OBJECTIVES: Amygdaloid nuclei and the hypothalamic structures are known to have intimate functional relationships. But to date the nature of this relationship has not been completely understood. In the present study, this relationship was evaluated. METHODS: Lesions were performed in basolateral nucleus of amygdala (BLA) and the ventromedial hypothalamus (VMH) in the same rats sequentially to evaluate both the individual, as well as combined effects of lesions of these two centres. Food intake, water intake and the body weight were studied before and after the lesions. RESULTS: The first lesion of BLA or VMH increased the food and water intake significantly (P < 0.01). But in the VMH lesioned rats, further lesioning of BLA, reduced the intensity of the hyperphagia. This suggested a kind of interrelationship between these centres, pointing out that intact BLA was instrumental in the development of VMH induced hyperphagia. To further assess the interactions of these two centres, three bottle free choice taste preference tests were undertaken by using sweet, salty solution along with tap water. The lesion of the BLA increased the intake of sweet tasting saccharin solution. This preference was retained even after the lesion of the VMH in the same rats. INTERPRETATION & CONCLUSION: Therefore it appeared as if the VMH neurons might not be involved in the sweet taste preference following BLA lesion, suggesting involvement of some other pathway for taste selection responses. But the development of full fledged VMH lesion induced hyperphagia could occur only in the presence of intact BLA. PMID- 11037682 TI - No correlation between primary mandibular anterior crowding and vertical craniofacial configuration or lower incisor inclination. AB - The aim of the present study was to evaluate the correlation between primary mandibular anterior crowding and vertical craniofacial configuration or sagittal lower incisor inclination. The collective comprised 100 consecutive patients (50 males, 50 females) subject to 4 selection criteria: mandibular anterior crowding > or = 1.0 mm, no previous orthodontic therapy, completely preserved deciduous canines and molars, and lateral cephalograms of good diagnostic quality before initiation of orthodontic treatment. Study casts of all patients were used to measure the mesiodistal width of the incisors and deciduous canines as well as the anterior arch circumference. The difference between the 2 represented the amount of crowding. On all cephalograms 16 parameters of vertical craniofacial configuration and 9 parameters of lower incisor inclination were measured. For error evaluation all measurements were repeated independently on 10 randomly selected study casts and cephalograms. This error amounted for the combined tooth width to 0.51% +/- 0.81, for arch circumference to 0.03% +/- 0.68, and for all cephalometric parameters to 0.38% +/- 4.81. For all parameters, arithmetic means, standard deviations and ranges were calculated. Further, correlation coefficients were calculated between anterior crowding and all 25 cephalometric parameters. The arithmetic mean of crowding was m = 2.0 +/- 1.3 mm with a range of 1.0 to 6.6 mm. The values of all cephalometric parameters were close to well accepted norms in the literature. The correlation coefficients between crowding and all cephalometric parameters varied from r = 0.0 to 0.3. According to this study there is no correlation between primary mandibular anterior crowding and vertical craniofacial configuration or sagittal lower incisor inclination. PMID- 11037683 TI - Investigations on the palatal rugae pattern in cleft patients. Part I: A morphological analysis. AB - The characteristics of the palatal rugae zone (number of rugae, relief type, posterior limitation) were investigated on the maxillary casts of 44 patients with unilateral cleft lip and palate and 28 patients with bilateral clefts by means of reflex microscopy, a three-dimensional, computer-assisted, touch-free measuring system for the metrical registration and analysis of the parameters directly on the maxillary casts for the segments of the 2 cleft groups. The features "number of palatal rugae" and "relief type" (primary rugae) were determined both before and after surgical repair of the cleft palate. Both segments in unilateral cleft lip and palate and both lateral segments in bilateral clefts most commonly had 4 to 5 palatal rugae. The number of rugae in cleft patients is thus in a range that other authors have reported for non-cleft individuals. Following palatal cleft repair, the rugae counts per segment decreased significantly in patients with unilateral and bilateral cleft lip and palate but the 3rd rugae was never lost after surgery. The relief type identified in unilateral and bilateral cleft lip and palate was the same as in isolated cleft palates and did not differ from that in non-cleft subjects. The posterior limitation of the palatal rugae zone was determined both in a tooth-defined manner and as an absolute linear distance (at all time points). The most frequent tooth-defined posterior limitation of the rugae zone in unilateral and bilateral clefts was the second deciduous molar, which is also the position identified for non-cleft individuals. The linear distance from the tuberosity line to the rugae zone increased in all segments of unilateral and bilateral clefts during the interval up to palatal cleft repair, indicating sagittal maxillary development in the posterior area of the palate. Surgical repair of the cleft palate resulted in a significant shortening of the distance in both segments of the unilateral cleft, most likely due to the displacement of mucosa and periosteum required to cover the palatal cleft. PMID- 11037684 TI - Dental age in southwest Germany. A radiographic study. AB - A cross-sectional study was undertaken in order to assess the dental age of healthy Southwest German boys and girls between the ages of 2 and 20 years by evaluating 1,003 panoramic radiographs. Dental age was assessed according to the method of Demirjian et al. All permanent teeth of the lower left jaw except the third molar were rated, the development of each tooth was divided into 8 defined stages. Statistical evaluation revealed a correlation between the parameters chronological age and score sum of r = 0.85 for girls and r = 0.89 for boys. The values of the score sum in relation to chronological age were distributed as in a logistic function. Two gender-specific equations for calculating dental age were devised and a marked sexual dimorphism was found. With the beginning of root formation, the girls showed accelerated development. The fact that dental age distribution in Southwest Germany is not significantly correlated to that of a French-Canadian collective underlines Demirjian's demand for regional standards of dental development to be calculated using his score system. PMID- 11037685 TI - The need for treatment and satisfaction with dental appearance among young Finnish adults with and without a history of orthodontic treatment. AB - The aim of this investigation was to evaluate orthodontic treatment need and patient satisfaction among young adults living in a city where free-of-charge orthodontic treatment was provided. A total of 281 18- to 19-year-old subjects randomly selected from the population register of the city of Vantaa took part in the study. The drop-out rate was 30%. Treatment need was clinically assessed according to the Index of Orthodontic Treatment Need (IOTN), consisting of a Dental Health Component (DHC) and an Aesthetic Component (AC). Information on previous orthodontic treatment was based on the patient records. Satisfaction of the subjects with their dental appearance and with the orthodontic treatment received was obtained using a questionnaire. The rate of orthodontic treatment among the subjects was 46% (54% for the females and 37% for the males, p < 0.05). 4% had discontinued treatment. A definite need for treatment (DHC 4 to 5/AC 8 to 10) was assessed in 15% of the subjects, and borderline/moderate need (DHC 3/AC 5 to 7) in 36%. No difference in IOTN scores between the treated and untreated subjects was found. Females had significantly more often no treatment need (DHC 1 to 2/AC 1 to 4) compared with males (p < 0.05). The majority of subjects (89%) reported that they were very or quite satisfied with their dental appearance. The odds of being satisfied were significantly higher for the treated subjects (OR = 2.71, p < 0.05) and lower for those at the non-attractive end of the AC scale (OR = 0.14, p < 0.01). Neither gender nor DHC grade significantly affected the odds of being satisfied among the subjects. The results indicate that the majority of young adults in this study were satisfied with their dental appearance regardless of objective treatment need of various degrees. The high treatment rate in relation to unnoticed treatment need calls for reevaluation of priorities in patient selection. PMID- 11037686 TI - Biomechanical investigation of the hybrid retraction spring. AB - We succeeded in developing a retraction spring that shows virtually constant retracting forces up to an activation of about 4.5 mm through the use of various non-linear materials. Compared to all other known retraction springs, an actual bodily retraction is possible over a large range for the first time. The clinical application requires no more than one reactivation. The anti-tipping moment is 10 Nmm and is to be considered constant over the entire activation range. This anti tipping moment produces an extrusive force for the canine and an intrusive force for the molar. This side-effect can be avoided by bending a sweep into the steel portion or compensated by bending a step into the steel portion of this retraction spring. The anti-rotational moment is about 3 to 5 Nmm measured over the entire activation range. In contrast to many other springs, the favorable M/F ratio for the anti-tipping movement allows an actual bodily retraction of canines. Even when the retracting force is no longer active, the moment that moves the root of the canine distally is still acting, so that the spring can also be used for the root movement. The M/F ratio for the anti-rotational movement is between 3 and 5 mm and therefore allows retraction of the canine without causing major distortions. Customary brackets with a .018" or .022" horizontal slot can be used, as the spring is designed for a .018" x .018" vertical slot. Each spring can be used for both the left and the right canines. The steel portion allows second-order and, if desired, third-order bends to be made. The additionally necessary bends have already been described. The hybrid retraction spring can also be applied for en masse retraction of incisors if a cross-tube is used for the anterior area. PMID- 11037687 TI - Bonded retainers--clinical reliability. AB - Bonded retainers have become a very important retention appliance in orthodontic treatment. They are popular because they are considered reliable, independent of patient cooperation, highly efficient, easy to fabricate, and almost invisible. Of these traits, reliability is the subject of this clinical study. A total of 549 patients with retainers were analyzed with regard to wearing time, extension of the retainer, mean time between failures, operator, and age of patient. The average frequency of breakage or loss was 0.55 per retainer per year. This frequency was dependent primarily on the operator who bonded the retainer and on the extent of the retainer. If the upper canines were involved, reliability was lower. The majority of failures occurred during the first 3 to 6 months. The study showed that bonded retainers represent a highly efficient and reliable retention appliance suited to long-term use. PMID- 11037688 TI - Lingual orthodontics (Part 4): Economic lingual treatment (ECO-lingual therapy). AB - One key factor in the economic lingual treatment concept presented in this series of articles is the use of prefabricated, customized nickel titanium arch wires with superelasticity and temperature-dependent spring-back qualities (memory effect). Using a modern, e-modulus driven archwire sequence, a non-extraction case can be treated with 1 or 2 wires per arch, and an extraction case with 2 or 3 wires per arch. This article, the last in the Lingual Orthodontics series, uses clinical examples to show the various clinical potentials of these wires with temperature-dependent superelasticity, as not only part of but occasionally all the treatment is done with these prefabricated, customized nickel titanium wires. PMID- 11037689 TI - Report on the 12th symposium of the Interdisciplinary Research Group on Cleft Lip and Palate, 29th/30th October, 1999. PMID- 11037690 TI - Effect of wearing clothes on oxygen uptake and ratings of perceived exertion while swimming. AB - For a comparative study between swimming in swimwear (control-sw) and swimming in clothes (clothes-sw), oxygen uptake (VO2) and ratings of perceived exertion (RPE) were measured. The subjects were six male members of a university swimming team. Three swimming strokes--the breaststroke, the front crawl stroke and the elementary backstroke--were applied. With regards to clothes-sw, swimmers wore T shirts, sportswear (shirt and pants) over swimwear and running shoes. In both cases of control-sw and clothes-sw, the VO2 was increased exponentially with increased swimming speed. The VO2 of the subjects during the clothed tests did not exceed 1.4 times of that in the case of control-sw at swimming speeds below 0.3 m/s. As swimming speeds increased, VO2 difference in both cases increased. Consequently, VO2 in the clothed tests was equal to 1.5-1.6 times and 1.5-1.8 times of that in the swimwear tests at speeds of 0.5 and 0.7 m/s, respectively. At speeds below 0.6 m/s in clothes-sw, the breaststroke showed lower VO2 than the front crawl stroke, and the elementary backstroke showed higher VO2 than the other two swimming strokes. RPE increased linearly with %peak VO2. In addition, any RPE differences among the three swimming strokes were not shown in the control-sw tests. At an exercise intensity above 60 %peak VO2, clothed swimmers showed slightly higher RPE in the front crawl stroke compared to that in the two other swimming strokes. PMID- 11037691 TI - Influence of water exercise and land stretching on salivary cortisol concentrations and anxiety in chronic low back pain patients. AB - Land stretching exercises are common exercise therapy for low back pain (LBP) patients. However, recently, water exercise became a popular rehabilitation for LBP patients, and many studies have reported the physical benefits of water exercise. This study compared the psychological and endocrinological effects of water exercise and land stretching by measuring salivary cortisol concentration and anxiety in chronic LBP patients. Seven volunteers (4 female and 3 male, mean age: 61.9 +/- 11.8 yrs) who suffered from chronic LBP (pain duration: 4.5 +/- 1.3 yrs) participated in the sessions of water exercise and land stretching programs (90 minutes) on different days. The land stretching program consisted mainly of stretching, and the water exercise program contained not only stretching, but also walking, jogging, muscle strengthening, swimming and relaxation. After both exercise programs, the subjective pain scores of the patients showed a significant decrease. Salivary cortisol concentrations were also significantly decreased during pre- to post-90 minute water exercise. (P < 0.05). With land stretching, salivary cortisol concentrations also decreased significantly (P < 0.05). State anxiety decreased significantly (P < 0.05) after both water exercise and land stretching compared with pre-exercise scores (P < 0.05), though no significant changes were found in the patients' trait anxiety scores. No significant correlation was found between salivary cortisol concentrations and state anxiety with water exercise and land stretching. The findings of the present study suggested both exercises showed similar tendencies, and had decreased salivary cortisol level and state anxiety. PMID- 11037692 TI - Effects of exercise experienced in the life stages on climacteric symptoms for females. AB - The purpose of this study is to investigate the effects of exercise experienced in the life stages on climacteric symptoms for females after menopause. Four hundred and eight postmenopausal women completed a questionnaire. The results were as follows: (1) Mean age at menopause +/- standard deviation was 50.1 +/- 0.5 and did not show a significant relationship with the degree of exercise in the life stages. (2) The degree of climacteric symptoms had a significant relationship, or a tendency toward a significant relationship, with the degree of exercise in and after the 40's; and the greater the degree of exercise, the lesser the degree of climacteric symptoms. (3) Kupperman's index was found to be, or tended to be, significantly related to the degree of exercise in and after the 30's. Those who exercised heavily in their 30's showed a significantly lower Kupperman's index. Those who answered that they had exercised "moderately" in their "40's to menopause" and "menopause to 60 years old" tended to have the lowest index. (4) Exercise experience in the life stages was negatively correlated, in particular, to psychosomatic symptoms among the 3 climacteric symptom categories. This negative correlation tended to be higher in those who answered that they had done "less exercise" in and after their 30's. (5) A significant relationship was noted between the degree of exercise in the 30's and "weakness" in Kupperman's index, and between exercise in and after the 40's and "nervousness" and "melancholia". Therefore, it is suggested that exercising "moderately" from the subjective viewpoint in the climacteric period may alleviate psychosomatic symptoms. PMID- 11037693 TI - Field studies on inhibitory influence of skin pressure exerted by a body compensatory brassiere on the amount of feces. AB - The present experiment investigated the effects of skin pressure produced by a body compensatory brassiere on defecation activity. Seven healthy females (11-41 yrs) volunteered as participants, being free of medication and constipation. The experiment lasted 3 weeks. The participants did not wear the body compensatory brassiere for the first week, wore it during waking hours for the second week, and again did not wear it for the third week. Whenever they desired to defecate, they did so and then weighted the amount of feces immediately by themselves. Eating times, daily amounts of foods and drinks, their menu, work intensity and its duration, retiring and rising time were controlled to be as similar as possible from day to day. The main finding was that the amount of feces was significantly smaller during the second week (wearing the body compensatory brassiere) than the first and third weeks (not wearing the body compensatory brassiere). These observations are discussed in terms of the suppression of the parasympathetic nervous system and intestine motility, and the delayed transit time in the large intestine. PMID- 11037694 TI - Physiological responses and RPE during underwater treadmill walking in women of middle and advanced age. AB - The purpose of this study was to examine the physiological responses and RPE during water walking using the Flowmill, which has a treadmill at the base of a water flume, in order to obtain basic data for prescribing water walking for people of middle and advanced age. Twenty healthy female volunteers with an age of 59.1 +/- 5.2 years took part in this study. They belonged to the same swimming club and regularly swam and exercised in water. Walking in water took place in the Flowmill. Subjects completed four consecutive bouts of 4 min duration at progressively increasing speeds (20, 30, 40 and 50 m/min) with 1 min rest between each bout. In addition, water velocity was adjusted to the walking speed of each bout. Subjects were instructed to swing both arms in order to maintain their balance during walking in water. The water depth was to the level of the xiphoid process and the water temperature was 30.31 +/- 0.08 degrees C. Both heart rate (HR) and oxygen uptake (VO2) increased exponentially as walking speed increased. HR was 125 +/- 15 bpm, and VO2 was 18.10 +/- 2.72 ml/kg.min-1 during walking in water at 50 m/min, which was the highest speed. The exercise intensity at this speed was equivalent to 5.2 +/- 0.8 Mets. The relationship between HR and VO2 during walking in water showed a highly significant linear relationship in each subject. There was also a highly significant linear relationship in the mean HR and VO2 of all subjects. Blood lactate concentration (LA) measured at rest and immediately after each bout was 1.1 +/- 0.4 mmol/l at rest, 1.0 +/- 0.2 mmol/l at 20 m/min, 1.0 +/- 0.3 mmol/l at 30 m/min, 1.1 +/- 0.2 mmol/l at 40 m/min, and 2.4 +/- 0.7 mmol/l at 50 m/min. LA at 50 m/min was significantly higher than at rest and at the other speeds. The relationship between HR and RPE during walking in water showed a highly significant linear relationship. The relationship between walking speed and energy expenditure calculated from VO2 and the respiratory exchange ratio (R) showed a high significant exponential relationship. These results suggested that HR and RPE can be effective indices for exercise prescription during Flowmill walking as with land walking. PMID- 11037695 TI - Effects of relative humidity on the population growth of house-dust mites. PMID- 11037696 TI - Preoperative exposure to calcium channel blockers suggests increased blood product use following cardiac surgery. AB - PURPOSE: In this study the authors reviewed the medical records of a random sample of patients undergoing coronary artery bypass grafting (CABG) during the preceding ten years at University Medical Center. The purpose of this study was to evaluate the impact of exposure to calcium channel blockers (CCB's) on blood product use following cardiopulmonary bypass (CPB). METHODS: DESIGN: Retrospective medical record review. SETTING: University hospital. PARTICIPANTS: 527 patients undergoing CABG or re-do CABG. INTERVENTIONS: The medical records of approximately 50% of patients undergoing CABG or re-do CABG at University Medical Center between 1988 and 1998 were randomly selected by the medical records librarian for review. Preoperative medications, bypass time and temperature, and blood product use were recorded. RESULTS: Of the 527 patients studied, 309 (59%) had no exposure to CCB's. 218 (41%) were on CCB's at the time of admission. Patients who were on CCB's had an average 12.5 (+/- 1.0) blood product units transfused following bypass whereas those not on CCB's had an average 8.7 (+/- 0.6) units transfused (p < 0.001). Use of packed red blood cells (p < 0.001), fresh frozen plasma (p = 0.018) and platelets (p = 0.023) were each individually significantly increased. CONCLUSIONS: In this study, it appeared that patients exposed to CCB's before cardiac revascularization received significantly more blood products than those who were not exposed to CCB's. Because of the limitations imposed by retrospective studies, further prospective studies are warranted to define the clinical significance of CCB use in the perioperative period. PMID- 11037698 TI - Med Web info-developing rules for the Web. PMID- 11037697 TI - Clonidine toxicity in an adolescent patient. AB - Clonidine is a central acting a2-agonist used primarily as an antihypertensive agent. Recently, it has been used for the treatment of attention deficit hyperactivity disorder in children and adolescents. When taken in excess, it can produce profound CNS depression, apnea, bradycardia and hypotension. A transient period of hypertension can sometimes occur. Treatment is primarily supportive, including respiratory support, atropine for bradycardia, and fluids and dopamine for hypotension. The CNS depression sometimes responds to naloxone. Young children are very sensitive to the toxic effects of clonidine. A case of an 11 year old adolescent who took an overdose of his clonidine is described to illustrate the toxicity of this agent. PMID- 11037699 TI - [The "open abdomen" as one of the possible therapeutic alternatives in postoperative complications]. AB - The authors discuss the important problem of healing, infection of surgical wounds and causes of healing per secundam. Despite efforts on the part of surgeons to close the surgical wound, in particular in old people with polymorbidity, hypoproteinaemia and anaemia, disintegration of the surgical wound may occur which cannot be closed and healing must proceed while the abdomen is open. PMID- 11037700 TI - [Gastroduodenal hemorrhage]. AB - Haemorrhage is a serious complication of peptic ulcer disease. It is an indication for urgent diagnostic and therapeutic endoscopy, which is at present the method of first choice. All patients with gastroduodenal peptic ulcer bleeding, who underwent endoscopy in Ist Department of Surgery in Bratislava between January 1995 and December 1999, were considered for retrospective study. A total of 291 patients (195 male and 96 female) underwent urgent endoscopy with a finding in 34.7% of patients with gastric ulcer and in 65.3% patients duodenal ulcer. The finding was Forrest I in 23%, Forrest II in 25.7% and Forrest III in 51.3% patients. Endoscopic hemostasis was used in 12.37% of patients. A first haemorrhage was found in 82.9% patients, a recurrent one in 17.1% patients. 41.5% of patients had positive peptic ulcer history. Surgical treatment was indicated in cases, when bleeding was not controlled by endoscopic means, or in cases of recurrent bleeding within 48 hours in 19 patients (6.52%). PMID- 11037701 TI - [Surgical treatment of acute diverticulitis of the large intestine]. AB - The authors analyze in this paper covering a ten-year period (1990-1999) a group of patients with acute perforated diverticulitis. The patients were classified according to Hinchey's classification. During this period 35 patients were operated with a high preoperative morbidity. With regard to the low number of complications of acute diverticulitis the authors do not prefer early preventive resections of non-complicated diverticulitis, but they tend to extend indications, first of all in cases of repeated recurrences, haemorrhage and in case of stenosis. In the IIIrd and IVth stage according to Hinchey's classification dominates Hartmann's operative procedure. But the authors agree with the trend to perform resection with primary anastomosis in the IIIrd stage, especially in cases of a good status of the patient and short duration of the disease. In cases of acute perforated diverticulitis operated in the first 24 hour interval the mortality rate was 23.1%. PMID- 11037702 TI - [Rectal cancer associated with irradiation--case report]. AB - The authors report one patient with rectal cancer after pelvic irradiation for gynaecological malignancy. The interval between irradiation and the diagnosis of rectal cancer was 31 years. Histological findings of radiation proctocolitis adjacent to the cancer were observed. PMID- 11037703 TI - [Malignant polyps of the colon and rectum]. AB - It is important for the general surgeon to have a clear idea of what is a malignant polyp and factors determining its clinical management. A malignant polyp is either sessile or pedunculated and harbors an invasive carcinoma which means that malignant cells have penetrated into or through the muscularis mucosae. Carcinoma in situ, intramucosal carcinoma, superficial carcinoma, carcinoma within the the mucosa or lamina propria are terms commonly used that must be distinguished from true invasive malignancy. These polyps should be treated by complete polypectomy and followed up as in a benign polyp. A polyp with invasive carcinoma requires careful evaluation to make a decision if simple polypectomy is sufficient treatment or whether surgical treatment is indicated. This decision is based on the risk factors such as a residual/recurrent tumour and the risk of lymph node metastases. We present a current review of the literature on the subject. PMID- 11037704 TI - [Biliary peritonitis]. AB - Biliary peritonitis is one of the infrequent types of acute abdomen. It is encountered in particular in older people and is associated with a high lethality. The diagnosis is difficult because the symptoms are not typical and frequently not very expressed. The importance of the disease is enhanced by the fact that a certain percentage of the cases develops as a complication and direct consequence of medical intervention. As several of these cases were encountered at our clinic in recent years and we had the opportunity to observe the "natural" course of the disease up to the lethal end in a female patient who refused all invasive diagnostic and therapeutic interventions, we present the clinical picture of the disease incl. auxiliary and laboratory examinations and the course of the disease. PMID- 11037705 TI - [Use of metal stents in treatment of benign stenoses of the biliary tract]. AB - OBJECTIVE: The authors submit for discussion a group of patients where they resolved a relapsing benign stenosis of the biliary pathways by insertion of a metal stent. MATERIAL AND METHODS: Since 1995 the authors introduced a metal stent in 29 patients with benign stenosis of the biliary pathways. In the majority they used a Palmaz stent. RESULTS: The 30-day mortality in the group of patients was 3.4% (1 patient, myocardial infarction). Two patients (6.9%) died 3 and 18 months resp. after insertion of the stent (pneumonia complicated by a pulmonary abscess, hepatorenal failure). Signs of obstruction of the stent (raised levels of bilirubin and liver enzymes) were observed in 5 patients (17%). In 8 cases (27%) migration of the stent occurred. The follow-up period in 26 patients with benign stenosis with a stent is two months--4 years (mean 19 months). Sixteen patients are followed up for more than 12 months. CONCLUSION: In the treatment, in particular of relapsing benign stenosis of the biliary pathways, specially when located in the area of the anastomosis, insertion of a metal stent is effective alternate treatment. PMID- 11037706 TI - [Dysphagia after the Rosetti fundoplication for gastroesophageal reflux]. AB - The authors describe as the possible cause of postoperative severe dysphagia after laparoscopic Rosetti's fundoplication low insertion of the cuff drawn behind the oesophagus to the anterior gastric wall. When the stomach is full the cuff is drawn into a spiral and the latter presses against the cardia and twists the cardia and abdominal oesophagus. For the condition unimpaired swallowing of the first mouthfuls is typical followed by severe dysphagia which can lead even to malnutrition. Common examinations may be normal except for the unaccountable dilatation of the oesophagus. An important role in detection of the cause is played by investigation of the passage of the barium suspension during swallowing with the patient in an upright position. The authors recommend insertion of the cuff as high as possible with careful mensuration of the permeability of the abdominal oesophagus during and at the end of surgery and fixation of the cuff to the diaphragm of diaphragmatic cruri. PMID- 11037707 TI - [Surgical treatment of fractures of the odontoid process]. AB - Forty-one cases of C2 odontoid process fractures, from a group of 273 surgically treated cervical spine trauma patients, were treated by anterior screw fixation during a 6-year period. There were 37 patients with Anderson and D'Alonzo type II. fractures and 4 with type III. The mean age of the patients was 52 years and the fracture was caused by falls in most cases. Medical complications were not observed during surgery, but in 3 patients, implant-related technical problems were recorded. In the 3-month follow-up, 2 patients died because of medical complications not related to the procedure. There were no other complications in the remaining patients. We included 21 patients in a 6-month follow-up. In 19 of them, the fracture healed (90.5%), while in 2 patients osteosynthesis failed, which required further stabilization. In the authors' opinion, direct osteosynthesis should be considered the method of choice in most C2 type II fractures as well as in some of type III fractures. PMID- 11037708 TI - [Repair of traumatic lesions in peripheral nerves--introduction]. AB - The reconstruction of traumatic lesions of peripheral nerves (PN) underwent the most significant improvement with the introduction of operating microscopes, microsurgical techniques and pertinent instruments, including microsutures. New information with respect to the pathophysiology of the degeneration and regeneration of peripheral nerves on a subcellular level also helped in the delineation of the rules and timing of optimal conditions for suturing. The present microtechnique seems to have reached its plateau with regard to capacity of available materials, technology and methodology. Despite this progress there is room for further improvement as addressed in this presentation. In this part of our work we would like to present a survey and analysis of the necessary theoretical knowledge of the basis of degeneration of a traumatized peripheral nerve at the level of molecular changes, as well as to indicate new technologies and materials. A detailed comparison of the classical method (microsuture) with the new technology (laser) has so far failed to demonstrate any significant difference. Therefore it appears necessary, when seeking advantages of the new methodology, to examine its application and not its quality, which will be the subject of the second part our work. PMID- 11037709 TI - Diagnostic importance of magnetic resonance angiography in arterial reconstructive surgery. AB - Magnetic resonance angiography (MRA) permits the non-invasive visualisation of blood flow in arteries and veins. It has an enormous potential in imaging of various vascular pathologies and it has been establishing itself as a reliable alternative to widespread contrast X-ray methods explaining morphology of vascular system. This advanced and safe diagnostic method is very rarely used in the clinical practice of vascular surgeons in Slovak republic. The aim of this report is to outline a clinical importance of MRA for arterial reconstructive surgery, its advantages, limitation as an imagine technique and its practical application in different anatomical areas of the human body which are related to operative activity of vascular surgeons. It is intended for briefly comparing MRA with other vascular imagine techniques more common used in clinical practice of our surgical and vascular surgery departments and outpatient clinics. PMID- 11037710 TI - [Injuries of the intracranial part of the carotid artery]. AB - The objective of the submitted work was to draw attention to different types of injuries of the intracranial portion of the carotid artery and some problems associated with its solution. The investigated group comprised 7 subjects (2 women, 5 men) aged 19 to 76 years who attended between Jan. 1, 1995 and Feb. 29 2000 the Neurosurgical Clinic in Kosice with sequelae of injuries of the intracranial carotid artery. Two patients developed a pseudoaneurysm. In one case it was manifested by subarachnoid haemorrhage on the 24th day after a crash, in one instance by diplopia six weeks after a retrobulbar injection. Injury of the intracavernous portion of the artery was manifested in one instance by profuse epistaxis on the 17th day after a fall from a bicycle, four times by the development of a carotid-cavernous fistula several days to 3 years after the head injury. In the diagnosis classical as well as MR and digital subtraction angiography were used. In the patient with epistaxis classical carotid angiography was 3 times negative. The results are comparable with data in the literature. In three patients the problem was resolved by ligature of the common carotid artery on the neck. In another three it was necessary to use extra intracranial trapping. One patient will be subjected to endovascular surgery. Two patients died (a 76-year-old woman from bronchopneumonia, a 19-year-old man from meningitis, despite a liquor fistula treated correctly by a patch). PMID- 11037711 TI - [An unusual contraindication for vascular reconstruction--case report]. AB - The authors report on an unusual cause of interruption of a planned vascular operation. The authors detected an anomalous biliary duct at the beginning of the vascular reconstruction operation. This duct was situated between the peritoneum and the abdominal fascia, and was unintentionally opened during the operation. Thus the laparotomy wound was contaminated by bile. To prevent contamination of the vascular prosthesis, which was planned for the reconstruction (bifurcation prosthesis), the operation had to be interrupted. PMID- 11037712 TI - The role of antithrombotic therapy on the patency of peripheral bypasses. AB - Peripheral bypass surgery is a well-established treatment for symptomatic atherosclerotic disease of the legs. To improve the long-term patency antithrombotic drugs and other adjuvant treatment are applied. In this review we summarize the results of randomized studies concerning antithrombotic treatment for prophylaxis after peripheral bypass surgery. Heparin is used routinely intra- and perioperatively without strong data, and according to few data low molecular weight heparin may be superior in this situation. Aspirin had positive effects in placebo-controlled studies after infrainguinal PTFE-bypasses, furthermore it is recommended due to its general cardiovascular risk reduction. Ticlopidine improved long-term patency in one randomized study after venous bypass surgery. Otherwise there are some data that long-term oral anticoagulation may be preferable in patients after venous bypass surgery. PMID- 11037713 TI - The transitional zone in the tunica media of renal arteries has a maximal length of 10 millimetres. AB - BACKGROUND: The goal of our study was to demonstrate and to determine the length of the transitional zone in the tunica media in renal arteries. The majority of renal artery atherosclerotic stenotic lesions occurs in this segment. PATIENTS AND METHODS: Anatomical and histological studies were performed on 26 renal arteries from 13 adults at autopsy (mean age 61.6 years, range 33 to 87 years). RESULTS: In the macroscopical examination the right renal arteries (RRA) were longer with a median 53.8 mm (range 38 to 65 mm) than the left renal arteries (LRA) with a median 47.6 mm (range 35 to 63 mm), the circumferences were nearly the same: RRA 10.9 mm (range 5 mm to 15 mm) and LRA 11 mm (range 5 mm to 15 mm). Probes for histological examinations were taken from three different regions of each renal artery (origin, 5 mm and 10 mm distal to the origin). We observed a typical elastic arterial structure at the origin and muscular types at the distal 10 mm region. At the distal 5 mm region variable ratios of elastic tissue (ET) and smooth muscle cells (SMC) were found as follows: 15 arteries presented an equal ratio of EM:SMC, 7 arteries presented ET > SMC and 4 arteries presented ET < SMC ratios. CONCLUSIONS: In this study we confirmed that in renal arteries, a transitional zone (TZ) that is an arterial segment with transition from elastic to muscular type, does exist, involving the maximal length of 10 mm. Further studies on the impact of the biomechanical properties of the transitional zone as a potential localizing factor in renal atherosclerotic disease are justified. In addition, the complex biomechanical behavior of the TZ of the arterial wall should be taken into consideration when interventional procedures are planned. PMID- 11037714 TI - Changes in the extracellular matrix of the vein wall--the cause of primary varicosis? AB - BACKGROUND: Conflicting theories on the development of primary varicosis have led to the molecular biological investigation of the vein wall or, more accurately, of the extracellular matrix. It was the aim of this study to quantify matrix expression and to compare pathological changes in the vein wall with valve orientated staging of varicosis, in order to determine indicators of the primary cause of varicosis. MATERIALS AND METHODS: Three hundred seventy-two tissue specimens of greater saphenous veins were obtained from 17 patients with varicosities and categorised according to Hach stage and procurement site. The specimens were compared with 36 specimens collected from six patients without varicosities, incubated with fluorescence-stained antibodies for collagen 4, laminin, fibronectin and tenascin prior to being assessed with confocal laser scan microscopy. In addition, 22 vein specimens (16 varicose, 6 normal veins) serving as negative controls were investigated. RESULTS: Image analysis and statistical evaluation showed that compared with normal veins, varicose veins are associated with a significant increase in matrix protein expression for collagen 4, laminin and tenascin. A trend towards an increase in matrix expression was further observed for fibronectin. There was, however, no difference between varicose veins and clinically healthy vein segments inferior to a varicose segment. CONCLUSION: If the findings of the present investigation can be confirmed by other studies, alterations in the vein wall may be regarded as the primary cause of varicosis and valvular insufficiency as the result of these changes. PMID- 11037715 TI - Prostaglandin E1-therapy reduces circulating adhesion molecules (ICAM-1, E selectin, VCAM-1) in peripheral vascular disease. AB - BACKGROUND: It has been postulated that adhesion molecules (AM) may be involved in development and progression of human atherosclerosis. We examined whether prostaglandin (PG) E1 affects circulating levels of the AM (ICAM-1, VCAM-1 and E selectin) in peripheral vascular disease (PVD) patients. METHODS AND RESULTS: AM are significantly (p < 0.01) increased in PVD (n = 65) as compared to controls (n = 31). There was no influence of risk factors. 26 PVD-patients received 2 different schemes of PGE1-therapy (group A [n = 17]; 5 ng PGE1/kg/min x 6 h x 5 d x 4 wk; group B [n = 9]; 60 micrograms PGE1/2 h x 5 d x 2 wk). PGE1 decreases all the AM significantly (p < 0.01) using both therapeutic schemes. Stopping PGE1 therapy reverses values within about 4 weeks. Details on therapeutic regimens (dose, duration, route, etc.) and individual response still need to be assessed. CONCLUSION: Our results indicate that PGE1-treatment of PVD is associated with a significant benefit on circulating AM. These findings are in line with the described anti-inflammatory actions of PGE1 and may represent a further contributing factor to the great variety of beneficial actions of PGE1 on human atherosclerosis. PMID- 11037716 TI - Ambulatory pressure gradient in the veins of the lower extremity. AB - BACKGROUND: Neovascularization is an important cause of venous reflux recurrence after high ligation of the long saphenous vein. The pathogenesis of this phenomenon is so far obscure. It is possible that a hemodynamic factor--a pressure gradient between the femoral vein and the residual long saphenous vein- could be the trigger initiating the process of neovascularization. PATIENTS AND METHODS: Venous pressure measurements on eight patients with primary varicose veins were performed in the erect position in the insufficient long saphenous vein on the thigh. Mean pressures in the quiet standing position and ambulatory pressures were considered. By interrupting the saphenous reflux either distally or proximally to the point of measurement the pressure conditions either in the femoral or in the crural veins were simulated. RESULTS: With the tourniquet placed distally to the point of measurement, the venous pressure in the upper interrupted segment of the long saphenous vein (equivalent to the pressure in the femoral vein) remained uninfluenced during ambulation. In contrast, by interrupting the reflux proximally to the point of measurement, a marked decrease of the ambulatory pressure in the lower part of the long saphenous vein (equivalent to the pressure in the crural veins) was noted. CONCLUSIONS: A pressure difference occurs between the veins of the thigh and the lower leg during the activation of the muscle venous pump. This fact may explain the tendency of recurrencies of varicose veins after high ligation of the long saphenous vein as well as the initiation of reflux. PMID- 11037717 TI - The role of microcirculatory techniques in patients with diabetic foot syndrome. AB - BACKGROUND: Diabetic foot syndrome (DFS) is a frequent complication of long standing diabetes mellitus, occurring in 10 to 30 percent of all diabetics with a vital risk for the affected limb and high mortality rates. Macroangiopathy, diabetic polyneuropathy and infections are trigger factors for DFS. Recent results imply a pathogenic role of functional and structural microcirculatory changes. The exact role of microangiopathy and the value of microcirculatory diagnostic methods in DFS have not yet been defined. PATIENTS AND METHODS: 78 patients with DFS (28 type I, 50 type II diabetics, mean age 63 years) were evaluated with video capillary microscopy, transcutaneous partial oxygen tension (tcpO2) measurement and laser Doppler fluxmetry (LDF) at the forefoot of the affected leg at admission and after revascularisation. Mean hospital stay was 28 +/- 11.7 days. Patients were stratified according to the etiology of DFS in patients with neuropathic lesions, macroangiopathic ulcers and mixed neuropathic angiopathic lesions. RESULTS: All groups had impaired microcirculation, and significant differences between groups were found in respect to capillary density. Reactive hyperemia, LDF pattern and tcpO2 did not differ significantly. Microcirculatory examinations did not yield additional information to clinical and Doppler sonographic results. CONCLUSION: In clinical practice, the role of microcirculation evaluation techniques for diabetic foot syndrome is limited. PMID- 11037718 TI - Segmental manifestation of peripheral atherosclerosis and its association to risk factors. AB - BACKGROUND: To prove whether the aortoiliac, femoropopliteal or crural segments of the peripheral arteries might have a different sensitivity to a risk profile we did a statistical analysis of segmental peripheral atherosclerosis and risk factors. PATIENTS AND METHODS: In 132 patients (mean age 61 +/- 13 years) with peripheral arterial occlusions the arterial segments with occlusion or stenosis were angiographically documented: 17 had occluded or stenosed aortoiliac, 45 femoropopliteal and 25 crural arteries and 45 patients had multiple manifestations. Analysis of total cholesterol, HDL- and LDL-cholesterol, triglyceride, lipoprotein a, fibrinogen, uric acid, homocysteine, hematocrit, erythrocyte sedimentation rate, HBA1, IgG- and IgM-antibodies versus Cytomegalovirus, Herpes simplex-virus, Chlamydia pneumoniae and Helicobacter pylori were done and nicotine abuse, arterial hypertension and obesity were evaluated. RESULTS: Age of the patients had the strongest correlation with isolated segmental manifestation (p < 0.0001). Patients with isolated aortoiliac manifestation were younger than patients without this manifestation (54 +/- 9 years versus 62 +/- 13 years). Patients with isolated femoropopliteal manifestation were older than patients without this manifestation (66 +/- 11 years versus 58 +/- 13 years). None of the investigated risk factors showed a correlation with these age related differences. Independent from the age related differences for the nicotine abuse a p-value of 0.08 was estimated, but in smokers a diffuse manifestation was most frequent. CONCLUSION: There are age dependent differences of the prevalence of isolated aortoiliac or femoropopliteal atherosclerotic occlusions or stenosis. An association of these differences to a specific risk profile was not found. PMID- 11037719 TI - FloSeal: a new hemostyptic agent in peripheral vascular surgery. AB - BACKGROUND: Bleeding is a common and often severe side-effect in vascular surgery. The use of glue is widely accepted to achieve a dry surgical field. The application of sealant is limited when the surface is covered with blood. Aim of this study was to evaluate a new sealant (FloSeal) in patients undergoing vascular surgery. PATIENTS AND METHODS: Between June 1998 and July 1999 a total of 17 patients with peripheral vascular interventions was included in this investigation. Effectiveness was measured by bleeding severity prior and after application, time to hemostasis, amount of fusion matrix necessary for hemostasis, the potential need for additional hemostatic measures, or the need for reoperations to control the bleeding. RESULTS: In 15 out of 17 patients bleeding was controlled with FloSeal alone, two patients required further surgical or hemostatic treatment. There were no local or systemic complications after use of this product. CONCLUSION: FloSeal is an advantageous hemostatic tool. PMID- 11037720 TI - Tibioperoneal arterial lesions and critical foot ischaemia: successful management by the use of short vein grafts and percutaneous transluminal angioplasty. AB - BACKGROUND: In a substantial number of mainly diabetic patients isolated crural arterial lesions are found to be the underlying cause for severe ischaemic foot lesions. Without revascularisation, patients with this specific occlusion pattern will inevitably face major amputation. To attain limb salvage in this setting, since the early eighties short vein grafts were used to bypass the occluded infrapopliteal arteries. More recently, percutaneous transluminal angioplasty (PTA) was also attempted to avoid limb loss in selected patients. PATIENTS AND METHODS: Since May 1986 in 125 patients 130 autologous bypass grafts from the BK popliteal artery or the proximal tibioperoneal arteries to malleolar vessels were performed in the presence of extended crural arterial occlusions and critical foot ischaemia (rest pain 3, tissue loss 127). In another series in 89 limbs (rest pain 5, tissue loss 84) of 84 patients PTA was done to treat 168 focal stenoses of > 50% diameter reduction and 11 short occlusions in a total of 135 crural arteries. RESULTS: Using life-table analysis, primary and secondary cumulative patency rates for short vein grafts with distal graft origin were 90% and 98% at 30 days, 76% and 83% at one year and 46% and 49% at seven years, respectively. The corresponding limb salvage rates amounted to 95%, 80% and 63%. Initial complete or partial technical success after PTA of crural arteries could be obtained in 93%: The limb salvage rates achieved were 95% at 30 days, 82% at one year and 63% at six years. CONCLUSION: Our results suggest that--depending on the extent of lesions--both short vein grafts as well as PTA are successful complementary treatment modalities to avoid limb loss in predominantly diabetic patients with infrapopliteal artery disease and critical ischaemia. PMID- 11037721 TI - A multivariate analysis of factors affecting patency of femoropopliteal and femorodistal bypass grafting. AB - BACKGROUND: The most important factors that determine the outcome after femoropopliteal and femorodistal arterial reconstruction are still controversial. This report analysis the factors that determine the early and late patency of distal arterial reconstruction. PATIENTS AND METHODS: A retrospective analysis of patency after femorodistal arterial reconstruction with a new method for evaluation of angiographic runoff was performed for 336 arterial reconstructions. The different pre-, per- and postoperative risk factors were analysed in a Cox proportional hazards model. RESULT: The patency was significantly better for vein grafts in comparison to composite grafts and prosthetic grafts. It was 74% for vein, 46% for composite and 43% for prosthetic reconstructions, respectively, at 12 months after arterial reconstruction. The cumulative life table patency rate in extremities with good, intermediate and poor runoff was 62, 30 and 10%, respectively at 36 months. The patency rates for extremities operated on for claudication was significantly better than for extremities operated on for critical ischaemia. The multivariate analysis of different factors in a Cox analysis revealed that only the status of distal runoff, the graft material and the site of the distal anastomosis independently and significantly influenced the patency rates. CONCLUSIONS: A new model for evaluation of distal runoff proved to predict the patency rate of femoropopliteal and femorodistal arterial reconstructions reasonably well in this retrospective analysis. PMID- 11037722 TI - Continuous contralateral jugular acid-base and blood gas monitoring during carotid endarterectomy. AB - BACKGROUND: The aim of this study is to evaluate the intravascular multiparameter sensor Paratrend 7 (P7) for continuous acid-base and blood gas monitoring after retrograde jugular catheterization during carotid endarterectomy. PATIENTS AND METHODS: We studied 11 patients with history of smoking (72.7%), coronary artery disease (72.7%), hypertension (100%), diabetes mellitus (55.5%) and TIA's and/or nondisabling stroke (90.9%). The contralateral internal jugular vein was punctured retrogradely and the calibrated P7 sensor was introduced. The sensor was removed after surgery. The P7 provides continuous graphical display of pH, pCO2, and pO2, while temperature, oxygen saturation, HCO3 concentration and base excess are displayed numerically. RESULTS: Mean duration of carotid cross clamping was 17.0 +/- 6.2 min. Mean stump pressure was 50.2 +/- 12.9 mmHg. Intraluminal shunting was not used in any operation. All sensors were easily inserted. During clamping, pH became persistently more acidic (7.31 to 7.28; p < 0.05), pCO2 was elevated (44.7 to 49.8 mmHg; p < 0.05) while, in the majority of the patients, there was a non significant decrease in pjvO2/SjvO2. Declamping was followed by a short period of decrease of pH and elevation of pCO2 reminiscent of wash out phenomena. PjvO2 was significantly elevated (53.8 +/- 5.2 to 59.0 +/- 5.8 mmHg; p < 0.001) after the restoration of flow. In one case, P7 was diagnostic for unsuccessful endarterectomy. CONCLUSIONS: P7 is useful during carotid endarterectomy providing continuous and "on-line" information on brain metabolism. It is a simple and powerful technique, which should be further investigated. PMID- 11037723 TI - [Acquired naevus flammeus (Fegeler syndrome)]. AB - We report the development of an acquired nevus flammeus following a ski accident. This disorder was first described by Fegeler 1949, who reported the case of a 43 year-old soldier who acquired a nevus flammeus in the face following a cranial trauma. Since then, a number of similar case reports have been published. Differential diagnosis is discussed, such as the unilateral nevoid teleangiectasia syndrome, eruptive nevoid teleangiectasia and eruptive spider nevi. PMID- 11037724 TI - Further cases of CSF confirmed in East Anglia. PMID- 11037725 TI - Suspected adverse reactions, 1999. PMID- 11037726 TI - Setting a course for the future. PMID- 11037727 TI - Presentation of awards. PMID- 11037728 TI - Histopathological survey of neoplasms in flat-coated retrievers, 1990 to 1998. AB - Over the period from March 1990 to December 1998, veterinary surgeons in general practice were invited to submit tissues suspected of being neoplastic which had been removed from flat-coated retrievers. When possible, pedigree details were obtained from the owners. In addition, data were collected from flat-coated retrievers known to have suffered from a neoplastic condition and for which a histopathological report was available. A total of 1023 submissions was obtained from 782 dogs. These included 165 non-neoplastic lesions (16 per cent), 447 benign samples (44 per cent) and 411 malignant samples (40 per cent). Soft tissue sarcomas accounted for 55 per cent of the malignant samples (26 per cent of all tumour samples and 22 per cent of all submissions) with 63 per cent of them being diagnosed as undifferentiated. Carcinomas accounted for 20 per cent of malignant samples (8 per cent of all submissions). Of the benign tumours, cutaneous histiocytoma was the most common diagnosis (48 per cent of benign tumours, 25 per cent of all tumours and 21 per cent of all submissions). PMID- 11037729 TI - Patterns of parasitic nematode infection and immunity in dairy heifers treated with ivermectin in a sustained-release bolus formulation either at turnout or in the middle of the grazing season. AB - Twenty-eight Holstein-Friesian heifers, born the previous year and weighing between 130 and 310 kg, were allocated to one of two treatment groups by restricted randomisation, based on their initial weight. The heifers in group 1 were each treated with ivermectin in a sustained-release bolus formulation at turnout in April, and those in group 2 were each given an ivermectin bolus on July 10, 84 days after turnout. On that day the mean geometric worm egg counts of groups 1 and 2 were 0.4/g and 38.8/g, respectively, and they both had a mean plasma pepsinogen concentration of 0.59 iu/litre; in group 1, two of 14 faecal samples were positive for Dictyocaulus viviparus larvae, and in group 2 all 13 samples were positive; in group 1 eight calves were positive and three inconclusive for the presence of antibodies to D viviparus, and in group 2 the corresponding figures were 10 positive and two inconclusive; the mean liveweights of groups 1 and 2 were 274.4 kg and 262.8 kg, respectively. By December 4,231 days after turnout, the corresponding results were: mean geometric worm egg counts of 2.2/g and 0.5/g; one of 13 and none of 14 faecal samples positive for D viviparus larvae; 12 positive and two inconclusive and none positive and 10 inconclusive for the presence of antibodies to D viviparus; 214 days after turnout their mean liveweights were 361.1 kg and 358.3 kg. Although the patterns of parasitic nematode infection were different in the two groups during the grazing season, by the time they were housed both groups had achieved similar liveweights and showed evidence of an immune response to both D viviparus and gastrointestinal nematodes. PMID- 11037730 TI - Commercial slaughter methods used on Atlantic salmon: determination of the onset of brain failure by electroencephalography. AB - This study investigated the effect of exsanguination without prior stunning, or carbon dioxide narcosis followed by exsanguination, or percussive stunning or spiking the brain, on the time taken to abolish the visual evoked responses (VERS) of farmed Atlantic salmon. Only percussive stunning and spiking killed the fish immediately and the other two methods resulted in aversive reactions by the fish. The VERS were lost between 148 and 440 seconds after exsanguination without stunning, and between 300 and 554 seconds after carbon dioxide narcosis followed by exsanguination. During both these procedures the fish showed strong aversive behaviour. In contrast, percussive stunning and spiking the brain could result in the immediate loss of VERS and no aversive reactions from the fish if the stun was applied correctly. PMID- 11037731 TI - Ectopic adrenal tissue in the cow. PMID- 11037732 TI - Isolation of Mycoplasma columborale from a fly (Musca domestica). PMID- 11037733 TI - Successful medical treatment of laryngeal chondritis in cattle. PMID- 11037734 TI - Bethanechol-responsive bladder atony in a colt foal after cystorrhaphy for cystorrhexis. PMID- 11037735 TI - Origins of the CSF outbreak. PMID- 11037736 TI - Increase in PDNS diagnoses in the Netherlands. PMID- 11037737 TI - Dispensing review. PMID- 11037738 TI - Sheep transport case. PMID- 11037739 TI - Headache characteristics during the development of tolerance to nitrates: pathophysiological implications. AB - Recent studies suggest that nitric oxide (NO) plays an important role in nitrate induced headache and in spontaneous migraine attacks. Organic nitrates act as prodrugs for NO and headache is a predominant adverse effect of nitrates but often disappears during continuous treatment. Insight into tolerance to headache could lead to insight into vascular headache mechanisms in general. The specific aim of the present study was therefore to characterize the headache and accompanying symptoms during continuous nitrate administration until a state of tolerance to headache had developed. 5-isosorbide-mononitrate (5-ISMN) 30 mg three times daily was administered orally for 7 days in 11 healthy subjects in a double-blind, randomized placebo controlled cross-over design. Wash-out between periods was 14 days or more. Haemodynamic data from the present study were compared to the observed changes of headache over time. Headache during 5-ISMN was longer lasting and more severe compared to placebo (P<0.004). In 10 subjects the headache fulfilled the pain sub-criteria for migraine and in five subjects all diagnostic criteria for migraine without aura were fulfilled. Conversely, 20 min of intravenous infusion of glyceryl trinitrate caused a milder headache and no migraine. The present results therefore suggest that NO may elicit a migraine attack in many healthy subjects if a high enough dose is given for several hours. A close temporal association between the disappearance of headache and the attenuation of the 5-ISMN induced dilatation of the superficial temporal artery was observed. In contrast, tolerance in the middle cerebral artery already appeared after 24 h, which was earlier than the development of tolerance to headache. If vasodilatation is the cause of headache the results point to extracerebral arteries. However, cytotoxic and pain modulating central nervous system effects of NO, the time courses of which are unknown, may also play a role, involving both intra- and extracranial arteries. PMID- 11037740 TI - Induction of nitrate tolerance is not a useful treatment in cluster headache. AB - The aims of the present study were to investigate whether induction of nitrate tolerance is a useful treatment in cluster headache and to correlate any changes in attack frequency of cluster headache and nitrate-induced headache to the vascular adaptation during continuous nitrate administration. The results were compared to results obtained from studies of nitrate tolerance in healthy subjects. MATERIALS AND METHODS: 5-isosorbide-mononitrate (5-ISMN) 30 mg was administered orally three times daily for 4 weeks in nine sufferers of chronic cluster headache in a double-blind, randomized placebo-controlled cross-over design. Blood velocity in the middle cerebral artery was measured with transcranial Doppler and the diameters of the temporal and radial arteries were measured with high frequency ultrasound. The haemodynamic data were compared to changes in the frequency of cluster headache attacks and interval headaches over time. RESULTS: Tolerance was complete within 24 h in the middle cerebral arteries and after 7 days in the symptomatic temporal artery, while tolerance of the radial artery was not observed within this period. The time profiles of tolerance were almost identical to the time profiles observed in healthy subjects. A close temporal association between the disappearance of nitrate-induced headache and tolerance of the temporal artery was observed but tolerance had no effect on cluster headache attack frequency. CONCLUSIONS: Induction of tolerance to nitrates cannot be used to treat cluster headache. If pain is related to arterial dilatation the results point to extracerebral rather than cerebral arteries as the site of nociception. However, other peripheral and central pain-modulating effects of nitric oxide, the time courses of which are unknown, should also be taken into consideration. PMID- 11037741 TI - Comparison of rizatriptan 10 mg vs. zolmitriptan 2.5 mg in the acute treatment of migraine. Rizatriptan-Zolmitriptan Study Group. AB - The efficacy and tolerability of rizatriptan (MAXALT) and zolmitriptan (ZOMIG) were compared in a randomized, double-blind, double-dummy, stratified (on prior use of rizatriptan and/or zolmitriptan), placebo-controlled, single attack study in 766 patients. Rizatriptan tended to provide freedom from pain sooner than zolmitriptan (hazard ratio 1.26, P = 0.075), acting within 60 min following dosing. More patients were pain free at 2 h on rizatriptan than on zolmitriptan (43.2% vs. 35.6%, P=0.041), while headache relief at 2 h was similar (70.5% vs. 66.8%). At 2 h, fewer patients on rizatriptan had symptoms of photophobia (35.6% vs. 43.5%, P = 0.029) and nausea (25.2% vs. 32.5%, P=0.046), and more patients on rizatriptan had normal function (45.4% vs. 37.0%, P=0.025) than zolmitriptan. Headache recurred in 28% of patients taking rizatriptan, 29% taking zolmitriptan and 26% taking placebo. Both active treatments were effective compared to placebo and were well tolerated. The most common side-effects with rizatriptan were asthenia/fatigue, somnolence and dizziness, while the most common side-effects with zolmitriptan were asthenia/fatigue and dizziness. PMID- 11037742 TI - The influence of ergotamine abuse on psychological and cognitive functioning. AB - Migraine patients abusing ergotamine often have chronic daily headaches associated with tiredness, sleep and memory disturbances, and reduced general well-being. We quantified psychological and cognitive functioning in 12 migraine patients with and 12 without ergotamine abuse (> or = 5 days/week for > or = 6 months) and 12 healthy controls. Psychological functioning assessed by Symptom Checklist-90 (SCL-90) and Profile Of Mood State (POMS), was impaired in ergotamine abusers compared to healthy controls. Cognitive functioning divided into four domains: attention (critical flicker frequency analysis and mental control subscale of the Wechsler Memory Scale (WMS), speed of information processing (reaction time tasks and lexical decision tasks), memory (four subscales of the WMS) and cognitive flexibility (trailmaking test and WMS digits backwards), was impaired in ergotamine abusers in speed of information processing and cognitive flexibility. These differences disappeared after correction for total SCL-90 scores. In conclusion, ergotamine abuse is associated with high psychological distress but not with structural impaired cognitive functioning. PMID- 11037743 TI - Tolerability and efficacy of naratriptan tablets in the acute treatment of migraine attacks for 1 year. Naratriptan Long-Term Study Group. AB - OBJECTIVE AND DESIGN: This open-label study was conducted to evaluate the tolerability and efficacy of the 5HT1 agonist naratriptan with repeated use in the acute treatment of migraine attacks for 1 year. Four hundred and seventeen (417) migraine patients treated 15,301 migraine attacks over the course of the study. RESULTS: The results show that 84% of attacks treated with a single 2.5 mg dose of naratriptan were not associated with the occurrence of an adverse event. The percentage of attacks associated with an adverse event did not increase with number of doses used to treat a given attack (1 vs. 2) or duration of use (0-6 months vs. > 6-12 months). The only adverse events experienced in > 2% of attacks throughout the 1-year study were nausea (3% of attacks), hyposalivation (2% of attacks), and drowsiness/sleepiness (2% of attacks). Headache relief 4 h post dose was reported in a median 70% of moderate or severe attacks and a median 86% of mild attacks treated with naratriptan tablets 2.5 mg. The percentages of patients reporting headache relief did not diminish as a function of increased duration of treatment (0-6 months vs. > 6-12 months) or frequency of use (for > 36 vs. < 36 attacks). The mean number of tablets taken per attack was 1.2. A second naratriptan 2.5 mg tablet was taken for headache recurrence in a mean 16% (median 8%) of attacks. CONCLUSION: The results of this study demonstrate that naratriptan tablets 2.5 mg taken for acute migraine attacks over a 1-year period are well-tolerated and effective. PMID- 11037744 TI - Memory disturbances in migraine with and without aura: a strategy problem? AB - Cognitive defects in migraine have been reported by several authors. These findings however, are controversial. In this study we carried out an investigation on 14 patients with migraine with aura and 16 with migraine without aura according to the International Headache Society criteria. They were submitted to a comprehensive battery of neuropsychological tests. The patients were compared with a control group not significantly different as to age, sex and education. Migraine subjects showed impaired neuropsychological performances only on some cognitive tests. Both groups of patients did worse than the control group on visuo-spatial memory tasks, while only migraineurs without aura showed significantly impaired verbal memory performances. The memory defects, both on visuo-spatial and on verbal cognitive tasks, could depend on an impaired recall mechanism. These memory difficulties seem related to strategically and organizationally defective aspects of learning. PMID- 11037745 TI - Migraine prevalence in adolescents aged 13-15: a student population-based study in Taiwan. AB - To estimate the lifetime migraine prevalence in school adolescents aged 13-15 in Taiwan, we conducted a self-administered questionnaire survey in four sampled public junior high schools. Migraine was diagnosed according to the diagnostic criteria of the International Headache Society. A total of 4064 students (1983 boys, 2081 girls) completed the questionnaire (response rate 91.6%). The lifetime prevalence of migraine was 6.8%. It was significantly higher in girls than boys (7.8% vs. 5.7%) and increased with age in both sexes. Students with migraine were more likely to be absent from school because of their headaches than those with non-migraine headaches (30% vs. 14%, odds ratio (OR) 2.7). They were also more likely to use painkillers for their headaches than their non-migraine headache peers (72% vs. 40%, OR 4.0). These results suggest that migraine is a common disorder of adolescents in Taiwan and its impact on the quality of life can not be ignored. PMID- 11037746 TI - Central sensitization in tension-type headache--possible pathophysiological mechanisms. AB - The aim of the present thesis was to investigate the pathophysiology of chronic tension-type headache with special reference to central mechanisms. Increased tenderness to palpation of pericranial myofascial tissues is the most apparent abnormality in patients with tension-type headache. A new piece of equipment, a so-called palpometer, that makes it possible to control the pressure intensity exerted during palpation, was developed. Thereafter, it was demonstrated that the measurement of tenderness could be compared between two observers if the palpation pressure was controlled, and that the Total Tenderness Scoring system was well suited for the scoring of tenderness during manual palpation. Subsequently, it was found that pressure pain detection and tolerance thresholds were significantly decreased in the finger and tended to be decreased in the temporal region in chronic tension-type headache patients compared with controls. In addition, the electrical pain threshold in the cephalic region was significantly decreased in patients. It was concluded that the central pain sensitivity was increased in the patients probably due to sensitization of supraspinal neurones. The stimulus-response function for palpation pressure vs. pain was found to be qualitatively altered in chronic tension-type headache patients compared with controls. The abnormality was related to the degree of tenderness and not to the diagnosis of tension-type headache. In support of this, the stimulus-response function was found to be qualitatively altered also in patients with fibromyalgia. It was concluded that the qualitatively altered nociception was probably due to central sensitization at the level of the spinal dorsal horn/trigeminal nucleus. Thereafter, the prophylactic effect of amitriptyline, a non-selective serotonin (5-HT) reuptake inhibitor, and of citalopram, a highly selective 5-HT reuptake inhibitor, was examined in patients with chronic tension-type headache. Amitriptyline reduced headache significantly more than placebo, while citalopram had only a slight and insignificant effect. It was concluded that the blockade of 5-HT reuptake could only partly explain the efficacy of amitriptyline in tension-type headache, and that also other actions of amitriptyline, e.g. reduction of central sensitization, were involved. Finally, the plasma 5-HT level, the platelet 5-HT level and the number of platelet 5-HT transporters were found to be normal in chronic tension-type headache. On the basis of the present and previous studies, a pathophysiological model for tension-type headache is presented. According to the model, the main problem in chronic tension-type headache is central sensitization at the level of the spinal dorsal horn/trigeminal nucleus due to prolonged nociceptive inputs from pericranial myofascial tissues. The increased nociceptive input to supraspinal structures may in turn result in supraspinal sensitization. The central neuroplastic changes may affect the regulation of peripheral mechanisms and thereby lead to, for example, increased pericranial muscle activity or release of neurotransmitters in the myofascial tissues. By such mechanisms the central sensitization may be maintained even after the initial eliciting factors have been normalized, resulting in the conversion of episodic into chronic tension-type headache. Future basic and clinical research should aim at identifying the source of peripheral nociception in order to prevent the development of central sensitization and at ways of reducing established sensitization. This may lead to a much needed improvement in the treatment of chronic tension-type headache and other chronic myofascial pain conditions. PMID- 11037747 TI - Recurrent short-lasting headache associated with paroxysmal hypertension: a clonidine-responsive syndrome. AB - The clinical syndrome of hypertension, headache, palpitation, diaphoresis, flushing, and emotional lability is classically associated with pheochromocytoma. Two patients are presented with this constellation of symptoms in whom investigations for pheochromocytoma were unrevealing. Headache was the presenting and most prominent complaint, with daily episodes of short-lasting, intermittent, and paroxysmal attacks. Each paroxysm of headache was associated with a dramatic increase in systolic and diastolic blood pressure. After failure to control the labile fluctuations in blood pressure and headache with several classes of anti hypertensive medications used in combination, a prompt and persistent response occurred after the administration of clonidine. The pathophysiology of this syndrome and the mechanism of clonidine action are reviewed in the context of a possible failure of the baroreceptor reflex. PMID- 11037748 TI - SUNCT syndrome responsive to gabapentin (Neurontin). AB - A 48-year-old male suffering with SUNCT (severe unilateral neuralgiform headache with conjunctival injection and tearing, rhinorrhea and sub-clinical sweating) presented in 1996 after a 10-year history of multiple failed therapies. The symptoms included strictly left-sided ocular, as well as facial and temple pain. The pain attacks were burning, sharp, shooting and occurred 25 times daily, lasting 2 to 3 minutes with tearing and conjunctival injection. There was no associated nausea or vomiting, but there was photophobia. No other autonomic changes were reported and the pain was not triggerable. Initially Indocin (indomethacin) was tried without significant benefit. Gabapentin (Neurontin) was then started with improvement at 1800 mg per day. The patient was then lost to follow-up for 3 years, as he moved from the Los Angeles area. He returned in 1999 having stopped the gabapentin after his prescription ran out in 1996, reporting the pain returned immediately. Again gabapentin was prescribed and at 900 mg three times daily he has been pain free for 12 months. PMID- 11037749 TI - Cough headache associated with Chiari type-I malformation: responsiveness to indomethacin. PMID- 11037750 TI - Enamelin maps to human chromosome 4q21 within the autosomal dominant amelogenesis imperfecta locus. AB - Amelogenesis imperfecta is a group of hereditary enamel defects. Of the autosomal dominant forms, only the local hypoplastic type has been mapped to human chromosome 4q 13-4q21. Enamelin is a large enamel matrix protein secreted by ameloblasts. The purpose of this study was to determine the human chromosomal localization of enamelin to establish an association with various forms of amelogenesis imperfecta. Chromosomal mapping was performed by polymerase chain reaction (PCR) amplification using somatic hybrid and deletion/derivation cell line panels with an enamelin primer set based on 100% conserved regions between pig and mouse cDNAs. Sequence-tagged site content mapping using eight markers within the critical local hypoplastic amelogenesis imperfecta region was then performed using an isolated human enamelin genomic BAC clone. The human enamelin amplicon was confirmed by DNA sequence analysis, revealing 81% and 73% identity to pig and mouse cDNAs, respectively. PCR amplification using a somatic cell hybrid panel placed enamelin on chromosome 4 with analysis of a regional chromosome 4 mapping panel refining the localization to 4q 13.1-q21.23. An identified human enamelin BAC genomic clone was shown to contain markers D4S2604 and D4S2670, as well as the first exon of the human ameloblastin gene, placing enamelin in the critical amelogenesis imperfecta locus between markers HIS1 and D4S2604 at 4q21. Our results suggest that enamelin is a strong candidate gene for this disease. Furthermore, human 4q21 may contain a second cluster of enamel matrix genes located proximally to the identified cluster of dentin and bone genes. PMID- 11037751 TI - Craniofacial morphology, dental occlusion, tooth eruption, and dental maturity in boys of short stature with or without growth hormone deficiency. AB - The aim of this project was to study the craniofacial morphology, dental occlusion, dental maturation and tooth eruption in short-statured boys with growth hormone secretion ranging from low to high. The measurements from lateral and posteroanterior cephalograms, orthopantomograms and plaster models were used. Almost all linear measurements of the facial structures were significantly smaller. A disproportionate growth in the cranial base structures as well as in the jaws resulted in facial retrognathia, a proportionately smaller posterior than anterior facial height, and a steep vertical inclination of the mandible. Dental crowding was more common and the overbite was small. Dental maturity and tooth eruption were delayed 1.2 and 1.3 yr, respectively. No significant differences between the idiopathic short-statured and the growth hormone deficient group in any of the above-mentioned variables were found. It can be concluded that although most of the cephalometric variables measured differed significantly from the average, the facial appearance of the boys is not conspicuous and is of minor clinical importance. However, the short-statured boys might be in greater need of orthodontic treatment due to the higher percentage of dental crowding. PMID- 11037752 TI - Factors influencing proximal dental contact strengths. AB - The aim of this study was to systematically measure proximal contact strength in complete natural dentitions of 30 adults (25.3 +/- 3.0 years of age), and to analyze its relationship to tooth type, tooth location, chewing effort and time of day variation. The contact strengths were measured dynamically during removal of a calibrated 0.05-mm-thick metal strip between the proximal contacts of adjacent teeth. Proximal contact strengths were lower in the maxilla (2.51 +/- 1.36 N) compared to the mandible (4.26 +/- 1.88 N). Within the jaws, the lowest proximal contact strength was measured between canine and first premolar (2.91 +/ 1.79 N) and the highest between second premolar and first molar (3.73 +/- 1.95 N). Chewing increased the proximal contact strength within the maxilla (before: 2.51 +/- 1.36 N, after: 3.02 +/- 1.45 N) but it remained unchanged in the mandible (before: 4.26 +/- 1.88 N, after: 4.22 +/- 1.85 N). The proximal contact strength increased significantly from morning (3.39 +/- 1.86 N) to noon (3.61 +/- 1.77 N), and then decreased in the afternoon (3.43 +/- 1.60 N). It was concluded that proximal contact strength can be significantly influenced by location, tooth type, chewing and time of day variation. Based on the differences in distribution due to the effect of chewing and time of day, it is speculated that proximal contact strength is a physiological entity of multifactorial origin. PMID- 11037753 TI - Prevalence of hepatitis C virus in patients with lichen planus of the oral cavity and chronic liver disease. AB - Lichen planus (LP) may represent a mucosal reaction to a variety of factors including hepatitis C virus (HCV) infection. We compared the prevalence of HCV infection in patients with LP of the oral mucosa and chronic liver disease (LP CLD) with those suffering exclusively from LP or from chronic liver disease (CLD). A total of 267 outpatients participated in a prospective study. There were 41 patients in the LP-CLD group, 128 in the LP group, and 98 in the CLD group. The diagnosis of LP was based on typical macroscopic and histopathologic features and the diagnosis of liver disease on liver histology. Serum samples were screened for anti-HCV antibodies. In 89 patients, serum HCV RNA was also measured. The overall prevalence of anti-HCV antibodies was 29.2% (78/267 patients). Serum HCV RNA levels were positive in 96.2% of anti-HCV-positive patients and in none of anti-HCV-negative subjects. Anti-HCV-positivity was more frequent in the groups of LP-CLD (78%) and CLD (42.8%) than in the LP group (3.1%). It is concluded that HCV infection plays an etiopathogenetic role in CLD associated with oral LP, whereas according to the present findings, the majority of patients suffering exclusively from oral LP are not infected by the HCV. PMID- 11037754 TI - Differences in the composition of the subgingival microbiota of two periodontitis populations of different geographical origin. A comparison between Spain and The Netherlands. AB - The purpose of this study was to compare the subgingival microbiota of two geographically distinct patient populations using identical clinical and bacteriological methods. Adult patients with a diagnosis of periodontitis were consecutively selected according to pre-defined clinical criteria. Microbiological samples were taken from the deepest four sites with bleeding. The samples were plated on blood agar plates, for the determination of the total anaerobic counts and identification of specific bacterial pathogens, and on TSBV and McConkey for isolation of Actinobacillus actinomycetemcomitans and enteric rods, respectively. Thirty-one patients in Spain and 30 patients in The Netherlands were selected. Both patient groups showed similar clinical characteristics, both in terms of age, gender and periodontal clinical variables. A. actinomycetemcomitans was significantly more prevalent (23.3% vs. 3.2%) in the Dutch group, while Porphyromonas gingivalis was significantly more prevalent (64.5% vs. 36.7%) in the Spanish group. Bacteroides forsythus and most commensal periodontal pathogens showed similar prevalences, except Peptostreptococcus micros that was significantly more frequent in the Dutch group (96.7% vs. 74.2%). In summary, the subgingival microbiota from the Spanish group was characterised by a high prevalence of P. gingivalis and low of A. actinomycetemcomitans, while the flora from the Dutch group was characterised by a high prevalence of A. actinomycetemcomitans and P. micros. PMID- 11037755 TI - Automated immunofluorescence for enumeration of selected taxa in supragingival dental plaque. AB - The present study investigated a recently developed automated image analysis technique for its applicability to the enumeration of selected bacteria in supragingival dental plaque. Following initial calibration, the system is capable to count fluorescence-labeled target cells in up to 48 samples without user interference. Test samples contained a characteristic mixture of planktonic bacteria, small almost planar bacterial aggregates, and large, virtually indisruptable clumps with cells from multiple species. Due to their complex composition, these samples provided a challenging validation step for the image analysis system. Automated enumeration of target bacteria was compared with visual counting of the fluorescence-labeled bacteria. Results are shown for six taxa (Actinomyces naeslundii, Fusobacterium nucleatum, Prevotella intermedia/Prev. nigrescens, Streptococcus gordonii/Strep. oralis/Strep. sanguis, Strep. sobrinus, and Veillonella dispar/ V. parvula) with characteristic differences in abundance, cell morphology and aggregation behavior. Results revealed good correspondence between the two enumeration techniques (correlation coefficients ranging from 0.77 to 0.92) provided that the portion of target bacteria exceeded 0.05% of the total bacterial cell number. This work demonstrates the applicability and usefulness of fully automated immunofluorescence to analyze such complex ecosystems as supragingival dental plaque. PMID- 11037756 TI - Microbiological aspects of an in situ model to study effects of antimicrobial agents on dental plaque ecology. AB - This study validates an in situ model for ecological studies of dental plaque exposed to various antimicrobial agents with different modes of action on plaque bacteria. Eleven subjects wore two acrylic appliances, each containing two bovine enamel discs, during two 1-wk test periods. Using a split-mouth crossover design, the appliances were dipped twice daily for 1 min into water (control; treatment A), fluoride (26.3 mM NaF; B), zinc acetate (20.0 mM; C), or fluoride plus zinc acetate (D). Four of the subjects used also chlorhexidine diacetate (2.2 mM; E) and chlorhexidine plus fluoride (F). At the end of each period, plaque was collected from the discs, after which the microbiota were analyzed by culture, automated quantitative immunofluorescence, and a viability fluorescence stain. As compared to control, treatments B, C, and D resulted in a significant reduction of individual taxa as detected by immunofluorescence, whereas similar bacterial viability and total bacterial numbers were observed. In contrast, chlorhexidine significantly reduced bacterial viability, total cell numbers, and the abundance of most of the enumerated taxa. We conclude that this in situ model is well suited to study effects of antimicrobial agents on dental plaque ecology. Combined with viability testing, immunofluorescence is obviously superior to culture in detecting taxa-specific shifts caused by antimicrobial agents. PMID- 11037757 TI - Dental plaque mass and acid production activity of the microbiota on teeth. AB - With the purpose of elucidating the effect of dental plaque thickness on the acid production activity in dental plaque, we studied the prevalence of acid anions in dental plaque of children harbouring different amount of plaque on their teeth. On two occasions, the occurrence of plaque on the dentition was scored (Silness Loe's index), and plaque on available smooth surfaces was collected and the wet weight determined. On the first occasion, the amount of acid anions in resting plaque, and on the second occasion, the acid anions of sucrose-exposed plaque were analysed with isotachophoresis. Mean value of plaque wet weights per individual dentition was 11.7 +/- 9.0 mg (first occasion) and 11.1 +/- 8.9 mg (second occasion). Dominating anions in resting plaque were acetate < propionate < lactate and in sucrose-exposed plaque lactate < acetate < propionate. The microbial acid production activity (acid anion per mg wet plaque weight) decreased with increasing weights of the plaque mass. The findings illustrate the cellular glycolytic metabolism of plaque microorganisms in thin and thick plaque, being a consequence of the diverse environmental condition existing in these ecosystems. PMID- 11037758 TI - Film-forming properties and viscosities of saliva substitutes and human whole saliva. AB - Hypo-salivation, related to medical remedies, is an increasing clinical problem. Studies report a weak correlation between subjective mouth dryness and objective sialometry. This indicates that both quantity and quality of saliva are important for the surface-associated functions of saliva, such as lubrication and hydration, to be expressed. Film-forming properties and viscosities of three saliva substitutes were compared to human saliva. Adsorption to surfaces was measured by ellipsometry, infrared spectroscopy and drop-volume technique. Viscosity measurements were carried out using an oscillating rheometer. Saliva, with the lowest viscosity value and the highest protein content, presented superior film retention on both hydrophilic and hydrophobic surfaces. The carboxymethylcellulose-based MAS 84 showed intermediate values of viscosity, poorest ability to reduce surface tension, and negligible film-forming capacity. The porcine mucin-based Saliva Orthana showed about twice the viscosity of saliva and film-forming capability on preferably hydrophobic substrates. Salinum, a linseed extract, possessed the highest viscosity value and an initial surface tension close to that of saliva. The film retention on hydrophilic surfaces was not as effective as for saliva. The results indicate that the film-forming capacity of saliva substitutes is a property also to be considered in the exploration of clinically effective artificial salivas. PMID- 11037759 TI - Catch-up growth and craniofacial dimensions following administration of the antineoplastic agent vincristine to young rats. AB - Decreased linear growth is a common problem during childhood anticancer therapy. Although catch-up growth may occur, short stature is sometimes permanent. A macroscopic experimental study was performed to investigate the catch-up growth of body and craniofacial structures of growing rats subjected to the antineoplastic agent vincristine. A total of 150 Long-Evans/Turku strain rats were randomly divided into one of two treatment or control groups. A group of 10 d-old rats was given a single s.c. injection of 0.0375 mg/kg vincristine, while another treatment group of 30-d-old rats received three injections of 0.05 mg/kg vincristine. All rats were killed at the age of 100 d. Twelve different craniofacial dimensions, tibia and body length, and final weight were recorded. Female and male groups were analyzed separately. As an expression of catch-up growth, most of the body and craniofacial structures returned to the control level by 100 d of age. A clear sex-dependency was seen in the measurements which remained reduced so that females seemed to catch-up better than males. The timing of the first injection was found to be important for the catch-up growth in the neurocranial area. PMID- 11037760 TI - Bovine dental papilla-derived cells immortalized with HPV 18 E6/E7. AB - In vitro investigations of cell-specific metabolism and cell interactions as well as biocompatibility studies are often hampered by the limited lifespan of primary cells originating from target tissues like the oral mucosa, gingiva or pulp. Pulp cells, as do other primary cells, undergo senescence after several passages in vitro. However, senescence can be overcome by transfection of primary cells with oncogenes like the HPV 18 (human papillomavirus 18) E6/E7 oncogene, resulting in immortalized cell lines. The purpose of our study was to establish and preliminary characterize an immortalized bovine dental papilla-derived cell line by transfection with HPV 18 E6/E7 for future use in biocompatibility testing of dental materials. First passage dental papilla-derived cells from molar tooth germs of 6-month-old calves were transfected by electroporation with pUC18 LCR E6/E7 coding for the HPV 18 E6/E7 oncogene. Cells underwent crisis after 5 wk in culture. Distinct cell colonies arose after about 9 wk. Cells were cloned by single cell dilution in passage 15 and 17. Out of three cell clones maintained in culture, two cell clones showed cell death after 28 and 30 wk, respectively. One cell clone overcame a second crisis after 38 wk and was maintained in culture until passage 75. Stable gene expression of HPV 18 E6/E7 oncogenes was verified by polymerase chain reaction (PCR) and immunohistochemistry. Reverse transcription (RT)-PCR revealed that the established cell line (passage 50) expresses procollagen type I, alkaline phosphatase and osteocalcin. This suggests that the immortalization with HPV 18 E6/E7 results in a cell line, which maintained phenotypic characteristics of odontoblast-like cells. PMID- 11037761 TI - Release of prostaglandin E2, IL-6 and IL-8 from human oral epithelial culture models after exposure to compounds of dental materials. AB - A three-dimensional human tissue model based on TR146 cells isolated from a squamous cell carcinoma of the buccal mucosa was used to test for the release of the proinflammatory molecules prostaglandin E2 (PGE2), interleukin 6 (IL-6), and interleukin 8 (IL-8) after exposure to nickel chloride (NiCl2), cobalt chloride (COCl2), palladium chloride (PdCl2), and triethylene glycol dimethacrylate (TEGDMA). These compounds have documented adverse biological effects in vitro. The release of PGE2 from the tissue culture models was inversely correlated with cell viability (MTT assay). Toxic concentrations of NiCl2 and CoCl2 induced the release of PGE2 by factors of about 200-300 compared to controls, but PdCl2 which was nontoxic enhanced PGE2 levels about 10-fold. TEGDMA, however, did not stimulate PGE2 release. None or weakly toxic concentrations of Ni and Co chloride induced IL-6 and IL-8 release by a factor of 5-10 compared to controls. The amounts of IL-6 were induced 25- to 30-fold by PdCl2 under physiological conditions, and IL-8 levels were also slightly enhanced. Nontoxic TEGDMA concentrations induced IL-6 levels 5-fold, but IL-8 amounts increased only slightly. We conclude that a steep rise of PGE2 is closely associated with cytotoxicity. On the other hand, the specific induction of IL-6 occurs at much lower concentrations. Therefore, the measurement of this cytokine may be included as another parameter in evaluating the biological activity of dental materials under nontoxic experimental conditions in vitro. PMID- 11037762 TI - Does chemomechanical caries removal affect dentine adhesion? AB - The purpose of this study was to determine whether shear bond strengths (SBS) to chemomechanically excavated dentine (Carisolv) differed from the SBS after conventional caries removal (bur). The following adhesive systems were used: Dyract AP/NRC/Prime & Bond NT; Dyract AP/Prime & Bond NT; Tetric Ceram/ Syntac SC; Tetric Ceram/Syntac SC (no etch) (TC 2); Definite/Etch & Prime 3.0; Definite/experimental. One-hundred and twenty human molars with occlusal caries were assigned to 12 groups. Sequential caries removal was controlled with laser fluorescence. After water storage, the samples were tested in a single-plane shear test assembly. The results demonstrated differences between the bonding systems, whereas the mode of caries removal had no consistent effect. If total etch technique was neglected, the results of the composite resin (TC 2) indicated a tendency to higher SBS in the Carisolv group (18.6 +/- 4.6 MPa) compared with conventional treatment (14.1 +/- 3.9 MPa). The present data demonstrate that chemomechanical caries removal has no adverse effect on bonding of modern adhesive systems to dentine. Smear layer-dissolving or -modifying bonding systems could potentially benefit from chemomechanical pretreatment. PMID- 11037763 TI - Adhesive bonding of titanium with a thione-phosphate dual functional primer and self-curing luting agents. AB - A consistent bond between the metal framework and the luting agent is desired when resin-bonded prostheses are constructed with titanium. The purpose of this study was to evaluate three different metal primers on titanium bonding. Two sources of titanium (machined 99.9% titanium and cast Titan Ingot JS2) were used. Disk specimens were bonded with eight combinations of three primers and two luting agents (Panavia 21 and Super-Bond C&B), including two controls. Shear bond strengths were determined after 24-h water storage and after 10,000 cycles of thermocycling. Bond strengths were influenced by thermocycling, primer, luting agent and their combinations, but no significant differences were found between the machined 99.9% titanium and the cast ingot. The thione-phosphate dual functional primer (Alloy Primer) was comparable to the phosphate primer (Cesead II Opaque Primer) and the thiophosphate primer (Metal Primer II) for bonding the titanium metals examined. The most durable bond was obtained in three combinations of these primers and one luting agent (Super-Bond C&B). PMID- 11037764 TI - Gender-specific differences in temporomandibular retrodiscal tissues of the goat. AB - Healthy, adult, male and female goat temporomandibular retrodiscal tissues were characterized to determine if biochemical differences existed between the genders. RNA concentrations were not different between male and female retrodiscal tissues; however, the DNA concentration in female retrodiscal tissues was 82% greater than in male retrodiscal tissues. Collagen concentrations were significantly greater in male retrodiscal tissues, and this was reflected in significant gender differences of type I and III collagen concentrations. More specifically, male temporomandibular retrodiscal tissues contained 70% more type I collagen and 119% more type III collagen when compared to female retrodiscal tissues. These differences in collagens and DNA reflect a gender difference in temporomandibular retrodiscal tissue composition that underlies divergent biomechanical and neurophysiological properties. PMID- 11037765 TI - Is anesthesia caused by potentiation of synaptic or intrinsic inhibition? Recent insights into the mechanisms of volatile anesthetics. AB - Volatile anesthetics modulate synaptic (GABAA receptor-mediated) and intrinsic (K+ channel-controlled) neuronal inhibition. GABAA receptor activity is enhanced, leading to increased charge transfer and prolonged synaptic inhibition, and members of the two pore domain family of potassium channels are activated, leading to neuronal hyperpolarization and reduced excitability. These effects may underlie different components of the complex anesthetic state. PMID- 11037766 TI - The effects of midazolam and morphine on analgesic and sedative activity of ketamine in rats. AB - The aim of this study was to investigate possible interactions between the analgesic activity of ketamine (an N-methyl-D-aspartate antagonist), midazolam (a benzodiazepine derivative) and morphine using the tail-flick test in rats. Animals were treated s.c. with ketamine (1.0-10.0 mg/kg), midazolam (0.3 mg/kg), or morphine (0.6 mg/kg) alone. or in combination The strongest analgesic effect of ketamine was observed after 3.0 mg/kg. In higher doses no enhancement of ketamine activity were found. After morphine and ketamine (3.0 mg/kg) or morphine, midazolam and ketamine co-administration. higher antinociceptive effects compared to ketamine activity were found. Rats administered midazolam and ketamine (3.0 mg/kg) showed a decrease of the effect of ketamine analgesia, and the antinociceptive effect of the three-component mixture was lower than after co injection of morphine and ketamine. The interaction of these two compounds with ketamine (5.0 mg/kg) occurred in a different manner, because midazolam led to a strong enhancement of ketamine analgesia. After morphine and ketamine (5.0 mg/kg) administration, very weak increase of ketamine analgesia was observed. The results of this study allow better understanding of the alteration of the analgesic effects of low doses of ketamine under the influence of morphine and midazolam. PMID- 11037767 TI - Effect of ligation of spleen vessels on left ventricular function and coronary blood flow in dogs injected with scorpion venom. AB - Scorpion sting may cause myocardial dysfunction in human victims, probably by increased O2 demand and decreased O2 supply. In dog, scorpion venom (SV) causes no myocardial dysfunction. Myocardium is probably protected by "autotransfusion" of blood from the spleen to the circulation, increasing coronary blood flow (CBF) and O2 delivery. We hypothesized that ligation of spleen vessels prior to injection of SV in dogs would prevent the autotransfusion of blood, thereby causing myocardial ischemia due to decreased CBF, simulating the hemodynamic pattern of human envenomation. We studied cardiac output (CO), CBF, left ventricular (LV) O2 delivery and contractility in 11 dogs injected with 0.07 mg/kg of SV (Leiurus quinquestriatus). Ligation of spleen vessels was performed on 6 of the 11 dogs prior to SV injection. 15 min after SV injection CO had increased by 186% in control dogs, while ligation of spleen vessels completely prevented CO elevation (p<0.001). In both groups, however, LV dp/dt increased by 400% and dp/dt/p by 170% (p<0.001). CBF increased by 350% and 550% in the spleen and control groups (p<0.001) respectively. This was associated with elevation of diastolic blood pressure and a decrease in coronary vascular resistance. LV O2 delivery increased (p<0.05) in both groups. At 60 minutes there was a decrease in CO, stroke work, and LV end systolic pressure in both groups, while LV contractility remained above baseline. Scorpion venom injection in dogs causes an initial increase in CO by auto-transfusion of blood from the spleen. Prevention of the autotransfusion does not preclude increases in CBF, O2 delivery and LV contractility. PMID- 11037768 TI - Assessment of intestinal permeability in rats by permeation of inulin fluorescein. AB - The measurement of intestinal permeability is widely used to assess different aspects of mucosal barrier disorders and related disease states, and has been proposed for evaluation of disease activity. To provide a simple method for assessment of intestinal permeability, we examined the permeation of inulin fluorescein (InFl) in rat models of small intestinal injury and colitis. Small intestinal or colonic inflammation was induced by either i.p. administration of indomethacin or rectal administration of trinitrobenzene sulfonic acid (TNBS), respectively. For monitoring of intestinal permeability, InFl was administered orally or rectally to rats with small intestinal or colonic inflammation, respectively, and its level in blood was determined by the fluorescence intensity in the plasma. In small intestinal injury, InFl reached its peak in plasma 3 h after oral administration, while in colitis the InFl peak was reached 1 h after rectal administration. The highest permeability was observed at 72 h or 12 h after induction of small intestinal or colonic inflammation, respectively. In small intestinal injury the InFl permeation, as measured by its plasma level prior to sacrifice, was in agreement with intestinal damage evaluated after sacrifice. In colitis, the permeability at 12 h after induction of the disease correlated well with mortality. These findings demonstrate that InFl can be used as a novel, safe and easy-to-use probe for the evaluation of gut permeation and follow-up of gastrointestinal injury. PMID- 11037769 TI - Analysis of bioactive components in traditional Chinese medicines by molecular biochromatography with alpha1-acid glycoprotein stationary phase. AB - Molecular biochromatography with alpha1-acid glycoprotein (AGP) stationary phase was proposed to screen and analyse the biologically active components in traditional Chinese medicines (TCMs) with extracts from Radix Salviae miltiorrhizae as a tested sample. More than ten peaks were resolved based on their affinity to AGP. The effects of concentrations of acetonitrile, pH, concentration of inorganic salt and temperature on the retention behaviors of several major active components were also investigated, and it was found that the hydrophobic effect is the major contributor to retention. Tanshinone IIA was identified as one of the principal bioactive components, which is the marker for the quality control of Radix Salviae miltiorrhizae and a complicated remedy named YiXiTongMai. The amount of tanshinone IIA in Radix Salviae miltiorrhizae and YiXiTongMai determined by this method was 2.9 mg/g (net weight, RSD 4.9%, n=5) and 0.078 mg/g (net weight, RSD 2.5%, n=3), respectively. The possibility for fast differentiation of the TCM sources was also studied by the comparison of the fingerprint of chromatograms for eight typical TCMs on the AGP column. It was observed that different TCMs showed different fingerprint characteristics. Even for the same plant, Rhizoma cimicifugae from three different geographical sources, although there were common characteristics, distinct differences in types and concentrations of biologically active components were clearly observed. It was shown that molecular biochromatography was an effective and fast way for the analysis and screening of biologically active compounds in traditional Chinese medicines. PMID- 11037770 TI - The effects of sertraline and fluoxetine on anxiety in the elevated plus-maze test in mice. AB - The aim of this study was to evaluate the acute and chronic effects of two SSRIs (sertraline and fluoxetine) on anxiety by the elevated plus-maze test. Diazepam increased the time spent in open arms significantly whereas the anxiogenic m chlorophenylpiperazine (m-CPP) decreased the time significantly. Acute sertraline (10 mg x kg(-1)) and fluoxetine (20 mg x kg(-1)) treatment significantly decreased the time spent in open arms. Acute fluoxetine (20 mg x kg(-1)) treatment also decreased the total number of enclosed arm entries. Seven days sertraline treatment decreased the time spent in open arms, whereas 14 days fluoxetine treatment increased the time spent in open arms. These results show that acute administration of SSRIs may produce anxiogenic effects in the elevated plus-maze test. PMID- 11037771 TI - The presence of Trypanosoma cruzi antigenic deposits. PMID- 11037772 TI - The results of tick-bone encephalitis (TBE)-vaccination. PMID- 11037773 TI - Risk factors for Echinococcus granulosus infection: a case-control study. AB - Despite the importance of cystic hydatid disease worldwide, no case-control study evaluating the risk factors for Echinococcus granulosus infection has been published to date. Thus, we carried out a hospital study to quantify different risk factors associated with the disease in a province marked by a high incidence of hydatidosis (Soria, Spain). The study population was composed of 127 cases and 127 controls matched by sex, age, and residence. Odds ratios (ORs) for hydatidosis decreased inversely with size of place of birth and residence, and increased with the number of dogs and years of coexistence with them. The variable involving the possibility of dogs ingesting uncooked viscera or carrion proved to be of greater importance (OR = 3.99, 95% confidence interval = 1.94 8.20). Risk factors for hydatidosis traceable to the family environment are of greater relative importance than those attributable to working directly with livestock. No association could be found between ingestion of raw green vegetables and hydatidosis. PMID- 11037774 TI - Malaria and anemia in antenatal women in Blantyre, Malawi: a twelve-month survey. AB - Malaria and anemia are common in pregnant African women. We screened 4,764 Malawian women at first antenatal visits for malaria and anemia. A total of 42.7% had a malaria infection, which was more common and of higher density in primigravidae (prevalence = 47.3%, geometric mean = 332 parasites/microl) and teenagers (49.8%, 390/microl) than in multigravidae (40.4%, 214/microl) or older women (40.6%, 227/microl). However, 35% of gravida 3+ women were parasitemic. A total of 57.2% of the women was anemic (hemoglobin < 11 g/dl), with moderate anemia (7.0-8.9 g/dl) in 14.9% and severe anemia (< 7 g/dl) in 3.2%. Prevalences of malaria and anemia were highest in the rainy season. Women with moderate/severe anemia had higher parasite prevalences and densities than women with mild/no anemia. Logistic regression showed that age, season, and trimester of presentation were significantly associated with the prevalence of malaria, but gravidity was not. In this urban setting, age and season are more important than gravidity as predictors of malaria at first antenatal visit, and parasitemia is frequent in women of all gravidities. PMID- 11037775 TI - Invasive bacterial infections of children in a rural province in the central Philippines. AB - The etiology of invasive bacterial infections was studied among 956 Filipino children less than five years old who fulfilled the World Health Organization criteria for severe or very severe pneumonia or had suspected meningitis or sepsis. The most common invasive infections were due to Streptococcus pneumoniae (12 [1.3%]) and Haemophilus influenzae (12 [1.3%]); including four cases of pneumococcal meningitis and 11 cases of H. influenzae meningitis. Type 1 was the most common (six of the 12 isolates) of the pneumococcal serotypes. Serotypes/groups 1, 6, 14, and 23 accounted for 91.7% of the invasive isolates. The majority of the H. influenzae strains from blood (10 out of 10) and cerebrospinal fluid (6 out of 7) were type b. Almost all of the invasive S. pneumoniae (9 out of 12) and H. influenzae (11 out of 12) infections were seen before one year of age, which stresses the need to investigate early immunization of children for H. influenzae type b and S. pneumoniae, as well as maternal immunization to maximize the potential of immunoprophylaxis. PMID- 11037776 TI - Intestinal parasitic infections, with a special emphasis on cryptosporidiosis, in Amerindians from western Venezuela. AB - The prevalences of intestinal parasites and intensities of helminth infections were studied in two Amerindian villages in Venezuela. Single stool specimens were collected from 303 individuals from Saimadoyi and 130 from Campo Rosario. Wet mounts, iron-hematoxylin-stained smears, and formalin-ether concentrates were examined for the presence of parasites; modified Ziehl-Neelsen carbol-fuchsin staining of 10% formalin-preserved stool was used to identify Cryptosporidium parvum. Helminth ova counts were made using the standard smear egg count technique. Mixed infections (Campo Rosario = 69.9%, Saimadoyi = 71.6%) were frequent. Overall infection rates with one or more species (Campo Rosario = 79.2%, Saimadoyi = 95.4%; P < 0.01) and with any protozoans (Campo Rosario = 60.8%, Saimadoyi = 72.3%; P < 0.05) were high and predominant (P < 0.05) in Saimadoyi. Cryptosporidiosis was identified in 38 subjects (8.8%) in both villages; 60.6% were asymptomatic carriers. The mean egg counts of helminths were heavier in Campo Rosario (P < 0.05), which was probably due to the drastic reduction of their lands along with their low standard of living. This study documents the change of intestinal parasitism pattern and deterioration of the health of Amerindians by the process of acculturation. PMID- 11037777 TI - Long-term effects of a nationwide control program on the seropositivity for Trypanosoma cruzi infection in young men from Argentina. AB - Unselected nationwide cohorts of Argentine men 18 years of age summoned for military service were tested for antibodies to Trypanosoma cruzi each year from 1981 to 1993. After an initial screening using indirect hemagglutination test, the positive sera were retested by titrated indirect hemagglutination and immunofluorescence antibody tests at 39 laboratories or at the national reference center in Buenos Aires. Nearly 1.8 million men were examined for T. cruzi antibodies using the same standardized procedures under a quality assurance program. The prevalence of seropositivity for T. cruzi decreased significantly from 5.8% in 1981 to 1.9% in 1993, but the decrease was not homogeneous among provinces within each region or constant over time. Prior to the nationwide control campaign initiated in 1961-1962, 15 provinces had high (> 20%) percentages of houses with domiciliary infestation by Triatoma infestans bugs, which decreased to nine provinces in 1982, and four provinces in 1992. The observed decrease in the prevalence of seropositivity for T. cruzi may be mostly attributed to the spraying with insecticides to eliminate the domiciliary populations of Triatoma infestans. The lack of a sustainable triatomine surveillance program set a limit to the decrease of seropositivity rates and prompted a revised strategy based on community participation. PMID- 11037778 TI - Application of a polymerase chain reaction-ELISA to detect Wuchereria bancrofti in pools of wild-caught Anopheles punctulatus in a filariasis control area in Papua New Guinea. AB - Chemotherapy-based eradication programs are aimed at stopping transmission of Wuchereria bancrofti by its obligatory mosquito vector. This study compares one year post-treatment W. bancrofti infection rates of Anopheles punctulatus, the main vector of lymphatic filariasis in Papua New Guinea, using traditional dissection techniques and a polymerase chain reaction (PCR)-based ELISA of a parasite-specific Ssp I repeat. A total of 633 mosquitoes in 35 batches were dissected. Six batches contained W. bancrofti-infected mosquitoes, giving a minimum infection rate of 0.9%. This value was not different than the actual infection rate, which was 9 (1.4%) of 633 mosquitoes (P = 0.48). The DNA was extracted from 47 pools containing a mean of 13.2 mosquitoes per pool. A total of 621 mosquitoes were processed for the PCR-ELISA, including 486 caught by human bait and 135 by light trap, which included both dead and live mosquitoes. Of 23 pools of alcohol-preserved human-bait mosquitoes, seven were positive by the PCR ELISA, giving an infection rate identical to that obtained by dissection of individual mosquitoes (1.4%). The minimum infection rates for pools of light-trap mosquitoes found dead and alive were 2.7% (2 of 74) and 4.9% (3 of 61), respectively. These values did not differ from each other (P = 0.84), but the overall infection rate of light-trap mosquitoes was greater than that of mosquitoes captured by human bait (3.7% versus 1.4%; P = 0.09). These data indicate that the PCR-ELISA of a W. bancrofti Ssp I repeat using pools of mosquitoes is comparable to traditional dissection techniques for monitoring transmission intensity following introduction of mass chemotherapy. This approach may also be useful for rapid and cost-effective assessment of transmission in endemic areas where the frequency of overt lymphatic pathology is low. PMID- 11037779 TI - Active, multisite, laboratory-based surveillance for Cryptosporidium parvum. AB - Cryptosporidium parvum leaped to the attention of the United States following the 1993 outbreak in Milwaukee, Wisconsin, which sickened 400,000 people. Other outbreaks in the United States have been associated with drinking and recreational water, consumption of contaminated foods, contact with animals, and childcare attendance. Despite its public health importance, the number of people who become infected each year is not known. In 1997, active surveillance for C. parvum was added to the Foodborne Diseases Active Surveillance Network (FoodNet), a collaborative effort among the Centers for Disease Control and Prevention, selected state health departments, the U.S. Departments of Agriculture and Food and Drug Administration. During the first 2 years of surveillance, 1,023 laboratory-confirmed cases of cryptosporidiosis were detected in FoodNet (Connecticut, Minnesota, Oregon, and selected counties in California, Georgia, Maryland, and New York). The annual rate per 100,000 persons was 2.3. Sixteen percent of case-patients were hospitalized. A seasonal increase in case detection was noted in late summer among persons less than 15 years of age. These data represent the first active multistate ascertainment of laboratory-confirmed cryptosporidiosis cases and provide useful information on the burden of disease in the United States. PMID- 11037781 TI - Vector densities that potentiate dengue outbreaks in a Brazilian city. AB - To identify the critical vector density that potentiates dengue outbreaks in an endemic site and to identify obstacles to anti-dengue activities, we correlated a series of dengue outbreaks in a Brazilian city with the intensity of its anti vector source-reduction activities. The proportion of houses infested by vector mosquitoes correlated inversely with intensity of anti-mosquito interventions, and the vector population developed independently of rainfall. Local periods of drought promoted vector abundance in two ways: residents stored water in which vector mosquitoes could breed, and cholera outbreaks due to contaminated water diverted local health workers from routine anti-vector activities. One dengue outbreak became apparent to authorities more than two months after it commenced but would have been identified almost immediately had dengue-like disease in indicator hospitals been monitored. Active surveillance, therefore, offers a window of opportunity for promptly executed anti-dengue interventions. Source reduction measures that suppress vector infestations to less than 1% of houses effectively avert outbreaks of dengue. PMID- 11037780 TI - Contrasting functions of IgG and IgE antimalarial antibodies in uncomplicated and severe Plasmodium falciparum malaria. AB - Plasmodial infection results in a significant elevation of the blood concentrations of immunoglobulins including IgE. Two well-characterized groups of adult Thai patients with either uncomplicated or severe Plasmodium falciparum malaria were studied over a period of four weeks. The mean parasitemias were approximately three-fold higher in patients with severe malaria than in those with uncomplicated disease. The mean concentrations of both total IgG and IgG antiplasmodial antibodies tended to be highest in the group with uncomplicated disease while total IgE and IgE antibodies were higher in the group with severe disease. The IgE antibodies detected in approximately 65% of the patients were positively correlated to parasitemia. These results suggest that antiplasmodial IgG antibodies are involved in reducing the severity of P. falciparum malaria, while IgE antibodies may contribute to the pathogenesis of this infection. PMID- 11037782 TI - Risk factors, seasonality, and trends of cryptosporidiosis among patients infected with human immunodeficiency virus. AB - To determine risk factors, seasonality, and trends of cryptosporidiosis among human immunodeficiency virus-positive (HIV+) patients in the New Orleans area, data from the New Orleans component of the Adult/Adolescent Spectrum of HIV Disease Study (ASD) were analyzed. A total of 6,913 HIV+ patients > or = 13 years of age were enrolled in the ASD database between 1990 and 1998. After an average follow-up of 42 months, cryptosporidiosis had been diagnosed in 239 patients (3.5%). The risk of developing cryptosporidiosis was higher among patients with CD4+ cell counts < 100 x 10(6)/L, among those who ever developed an acquired immunodeficiency syndrome (AIDS)-opportunistic illness, and among patients < 35 years old compared with their counterparts. A slight increase in cryptosporidiosis cases occurred in the spring compared with other seasons, but the difference was not statistically significant (P > 0.05). The prevalence of cryptosporidiosis increased from 2.9% (n = 7) in 1989 to 20% in 1994 (n = 48) before decreasing to 5.3% in 1998 (n = 14). Since a fair number of cryptosporidiosis cases are still being reported in the New Orleans area after the introduction of highly active antiretroviral therapy, further studies are needed to provide insight into the existence of potential environmental sources of Cryptosporidium. PMID- 11037783 TI - Risk factors for Giardia intestinalis infection in agricultural villages practicing wastewater irrigation in Mexico. AB - This study assessed the risk factors for Giardia intestinalis infection in an agricultural population in Mexico. Exposure groups included 2,257 individuals from households exposed to untreated wastewater, 2,147 from a group using the effluent from a series of reservoirs, and 2,344 from rain-fed agricultural villages. Stool samples were collected from 6,748 individuals. Wastewater samples were tested for fecal coliforms/100 ml and Giardia sp. cysts/L. Untreated wastewater samples contained 10(8) fecal coliforms/100 ml and up to 300 Giardia sp. cysts/L. Hydraulic retention (3-7 months) in the reservoirs, however, provided an improved effluent quality (10(1)-10(4) fecal coloforms/100 ml and < or = 5 Giardia sp. cysts/L). Children 1-14 years of age had the highest prevalence of infection (20%). Data showed marginal associations between storing drinking water in unprotected containers and lack of facilities for feces disposal and the risk of infection (odds ratios [ORs] = 1.76 and 1.19, 95% confidence intervals [CIs] = 0.95-3.23, and 0.97-1.45, respectively). Individuals purchasing vegetables at the city market had higher rates of infection than those buying at the village shop (OR = 2.49, 95% CI = 1.00-6.17). No excess risk was found in individuals exposed to untreated wastewater compared with controls (OR = 1.07, 95% CI = 0.84-1.36); the group using reservoir water was not different from the controls (OR = 1.22, 95% CI = 0.94-1.58). No risk from agricultural activities was detected (OR = 0.83). This pattern of infection may be addressed by primary health care and wastewater treatment. PMID- 11037784 TI - Short report: a consideration of primaquine dose adjustment for radical cure of Plasmodium vivax malaria. AB - Relapse of Plasmodium vivax malaria following standard primaquine dosing has been reported from many areas, and more recently from sub-Saharan Africa. In this report we describe eight episodes (in five patients) of treatment failure in non immune Israeli travelers returning from Ethiopia. Retrospective calculation of the primaquine dose per kilogram of body weight for 23 treatment courses showed a lower total dose per kilogram in heavier patients. The mean calculated dose (95% CI) in the eight failed treatments was 2.5 +/- 0.3 mg/kg compared with 4.4 +/- 0.5 mg/kg in the 15 successful treatment courses. Weight-adjusted dosing regimens may prevent inadvertent subtherapeutic drug failure, and thus apparent primaquine failure. In these cases, no relapses were observed in those who received > 3.5 mg/kg. Consideration should be given to adjusting the dose of primaquine according to body weight. For those infected by strains from Ethiopia a dose > 3.5 mg/kg is preferable. PMID- 11037785 TI - The changing in vitro susceptibility pattern to pyrimethamine/sulfadoxine in Plasmodium falciparum field isolates from Kilifi, Kenya. AB - Two clinical trials that used Falcidin (Cosmos Ltd., Nairobi, Kenya), the antifolate combination of pyrimethamine/sulfadoxine (PM/SD), as treatment for non severe falciparum malaria in children at Kilifi, Kenya in 1987-1988 and 1993-1995 have presented an opportunity to assess in vitro the susceptibility trend of Plasmodium falciparum to PM and SD over time on the Kenya coast. The first set of isolates was collected prior to the introduction of PM/SD into the Kenya Medical Research Institute/Wellcome Trust Research unit while the second set was taken soon after PM/SD was introduced in the study area as the first-line treatment drug for uncomplicated falciparum malaria. In the first trial, 69 isolates collected before and after treatment of malaria with PM/SD were tested directly in the field for susceptibility to PM and SD using the standard in vitro micro test technique, with minimal levels of folate. In the second trial, 97 isolates similarly collected were adapted to culture, and tested as described elsewhere. In both studies, PM and SD susceptibility tests were done separately. There was a highly significant decrease (P < 0.01) in the in vitro sensitivity of P. falciparum isolates to PM and SD between the two trials. In the first trial, the isolates were either sensitive to both PM and SD or resistant to PM and sensitive to SD. During the second trial, isolates were either resistant to PM and sensitive to SD or resistant to both drugs. These results are important in estimating the useful therapeutic life (UTL) of PM/SD in this area and in identifying alternative antifolate drugs. PMID- 11037786 TI - A randomized, double-blind, parallel-group, comparative safety, and efficacy trial of oral co-artemether versus oral chloroquine in the treatment of acute uncomplicated Plasmodium falciparum malaria in adults in India. AB - In India, treatment of acute, uncomplicated Plasmodium falciparum malaria is becoming increasingly difficult due to resistance to chloroquine, thus there is a need for new antimalarial drugs. CGP 56697 (co-artemether), a new drug, is a combination of artemether and lumefantrine in a single oral formulation (one tablet = 20 mg of artemether plus 120 mg of lumefantrine). In a double-blind study, 179 patients with acute uncomplicated P. falciparum malaria were randomly assigned to receive either CGP (n = 89) given as a short course of 4 x 4 tablets over a 48-hr period or chloroquine (n = 90) given as four tablets (one tablet = 150 mg of chloroquine base) initially, followed by two tablets each at 6-8, 24, and 48 hr. Due to a death in the chloroquine group and a decrease in the chloroquine cure rate to < 50% (based on the blinded overall cure rate at that time), recruitment was terminated prematurely. CGP 56697 showed a superior 28-day cure rate (95.4% versus 19.7%; P < 0.001), time to parasite clearance (median = 36 versus 60 hr; P < 0.001), and resolution of fever (median = 18 versus 27 hr; P = 0.0456). This drug provides a safe, effective, and rapid therapy for the treatment of acute uncomplicated P. falciparum malaria. PMID- 11037787 TI - Studies of the neurotoxicity of oral artemisinin derivatives in mice. AB - Intramuscular injections of high doses of the oil-soluble antimalarial artemisinin derivatives artemether and arteether produce an unusual pattern of selective damage to brain stem centers in experimental mammals, predominantly those involved in auditory processing and vestibular reflexes. We have shown recently in adult Swiss albino mice that parenteral artesunate, a water-soluble derivative, is significantly less neurotoxic than intramuscular artemether in this murine model. Using the same model, in which the drugs were administered daily for 28 days, the neurotoxic potential of the oral drugs was assessed and compared with the parenteral routes of administration. The dose causing neurotoxicity or death in 50% of animals (ED50), was approximately 300 mg/kg/day of oral artemether and artesunate compared to 50 mg/kg/day of intramuscular artemether. Doses of intramuscular artemether > 100 mg/kg/day were uniformly lethal. When oral artemether was given in peanut oil there was an increase in neurotoxicity and mortality compared with the aqueous suspension (P = 0.002), and when the food pellets were coated with artemether in oil, giving relatively constant oral intake, neurotoxicity was further increased; ED50 = 150 mg/kg/day (P = 0.017). These data indicate that once-daily oral administration of artesunate or artemether is relatively safe, presumably because the central nervous system is exposed transiently, whereas constant exposure either from depot intramuscular injection of oil-based drug, or constant oral intake carries relatively greater neurotoxic potential. PMID- 11037788 TI - Potential for New York mosquitoes to transmit West Nile virus. AB - We evaluated the potential for several North American mosquito species to transmit the newly introduced West Nile (WN) virus. Mosquitoes collected in the New York City Metropolitan Area during the recent (1999) WN outbreak were allowed to feed on chickens infected with WN virus isolated from a crow that had died during this outbreak. These mosquitoes were tested approximately 2 weeks later to determine infection, dissemination, and transmission rates. Culex pipiens mosquitoes were highly susceptible to infection, and nearly all individuals with a disseminated infection did transmit WN virus by bite. In contrast, Aedes vexans were only moderately susceptible to oral infection; however, those individuals inoculated with WN virus did transmit virus by bite. PMID- 11037789 TI - Density and spatial distribution of Ixodes pacificus (Acari: Ixodidae) in two recreational areas in north coastal California. AB - The density and distribution of Ixodes pacificus was assessed at 2 parks in north coastal California. The density of I. pacificus adults and nymphs varied significantly between years, trails, and sides of trails. Adult ticks occurred on vegetation along sun-exposed trails in January through March, their density (0 1.93 per 20 m) correlated with brush density, trail width, and presence of an uphill slope. Nymphs (0.06-5.10 per 20 m) occurred in leaf litter along shaded trails in May-July. Adult I. pacificus were rare at picnic sites (0.00-0.24 per 20 m), but nymphal densities (0.93-2.37 per 20 m) were comparable with those along some shaded trails. The prevalence of Borrelia burgdorferi in ticks (2.8% overall) did not differ significantly between locations, years, or stages. We conclude that the risk of acquiring Lyme disease in these sites is low, but varies among trails, seasons, and years. PMID- 11037790 TI - Further evidence for secretion of matrix metalloproteinase-1 by Meckel's chondrocytes during degradation of the extracellular matrix. AB - We examined the possibility that chondrocytes in Meckel's cartilage might secrete matrix metalloproteinase-1 (MMP-1) during degradation of the extracellular matrix. Evidence for the secretion of MMP-1 was obtained by immunohistochemical staining and immunoelectron microscopy, in addition to general histochemical staining for proteoglycans. Not only staining with toluidine blue and alcian blue but also immunostaining for chondroitin sulfate proteoglycan (CSPG) revealed that levels of glycoproteins are rapidly reduced at the late stage of degradation. MMP 1 was detected continuously in cells from chondrocytes at the early stage to hypertrophic chondrocytes at the late stage. Immunoelectron microscopy revealed that the deposition of colloidal golds shifted from an intracellular localization in chondrocytes at the early stage to pericellular spaces at the late stage. The localization of tissue inhibitor of the metalloproteinase-1 (TIMP-1) at the early stage was similar to that of MMP-1, but the level of TIMP-1 decreased significantly in hypertrophic cartilage. These findings suggest that MMP-1 is present continuously in Meckel's chondrocytes but that the active form, which degrades the extracellular matrix, is the MMP-1 that accumulates in the pericellular spaces around hypertrophic chondrocytes. PMID- 11037791 TI - Disruption of the olfactory placode and brain conditioned medium increase the number of LHRH immunostained neurons in explants. AB - The olfactory placode gives rise to both olfactory receptor neurons, which remain as a component of the peripheral nervous system, and to luteinizing hormone releasing hormone (LHRH) neurons, which migrate to the central nervous system. In this study, we used chick olfactory placode explants to ask several questions regarding LHRH neuronal differentiation. We found that explants of ectoderm from the fronto-nasal region of embryos as early as Hamilton & Hamburger (HH) stage 12 gave rise to LHRH neurons, that explants from all regions of the olfactory placode were able to generate LHRH neurons, that both brain conditioned medium and disruption of the olfactory placode increase the number of LHRH neurons observed in explants, and that the combination of these two manipulations results in the production of more LHRH neurons than either treatment alone. We conclude that LHRH neurons originate in the olfactory epithelium and that some of the same factors which influence olfactory receptor neuron development also affect LHRH neuronal development. PMID- 11037792 TI - Inhibition of nebulin and connectin (titin) for assembly of actin filaments during myofibrillogenesis. AB - In order to examine the role of cytoskeletal scaffolding proteins, nebulin and connectin (titin), in actin dynamics during myofibrillogenesis, rhodamine (rh) labeled actin was microinjected into cultured skeletal muscle cells in which the function of these proteins had been inhibited with their respective antibodies. In the nebulin function-inhibited cells, exogenously introduced actin formed irregularly distributed amorphous patches or bright foci inside the cells, but it was not incorporated into myofibrillar structures at any stage. Thus, the blockage of actin binding sites of nebulin seems to inhibit the association of actin monomers to the preexisting nebulin scaffold. In the cells inhibited with anti-connectin antibody, incorporation of rh-actin was similar to that in antibody-uninjected cells. These results support the idea that nebulin is related to the accessibility/exchangeability of actin into nascent myofibrils, but connectin does not have such a role in actin assembly. Since all antibodies recognizing different domains of nebulin filaments blocked actin incorporation along the entire length of actin filaments, inhibition of any domains of nebulin filaments seems to affect actin dynamics. PMID- 11037793 TI - Ultrastructure of the seminal vesicle of Phlebotomus perniciosus Newstead (Diptera, Psychodidae). AB - The seminal vesicles of Phlebotomus perniciosus were investigated by light microscopy, confocal scanning laser microscopy and by scanning and transmission electron microscopy. They have a complex structure, and three different morphological compartments called A, B and C are distinguished on the basis of their position and fine structure. Compartment A is continuous with the vasa deferentia and consists of a cylindrical wall limiting a lumen in which the spermatozoa are stored. Compartment B is hemispherical and surrounds compartment A like a muff. Compartment C constitutes an external coat surrounding A and B. The epithelial cells of each compartment are characterized by morphologically different secretory granules. The ultrastructural features of these cells are described and their role in sandfly reproductive biology is discussed. PMID- 11037794 TI - Differentiation of endothelial barrier function during normal angiogenesis requires homotypic VE-cadherin adhesion. AB - Endothelial cells express two principal cadherins: VE-cadherin and N-cadherin. We established previously that only VE-cadherin expression was increased during differentiation of barrier function by angiogenic endothelium of the chick chorioallantoic membrane (CAM). Presently anti-VE-cadherin mAb, applied to the CAM at day 4.5 of gestation, served to inhibit the abrupt reduction of macromolecular extravasation that occurs normally at day 5.0. Neither anti-N cadherin nor nonimmune IgG, on the other hand, prevented this temporal decrease of endothelial permeability. Despite the differential permeability responses, morphometric evaluations defined a reduction of mean paracellular cleft width after the application of either anti-VE-cadherin or anti-N-cadherin. Hence, alteration of molecular sieving characteristics within the junctional clefts, rather than modification of cleft dimensions; likely served as the principal modulator of macromolecular extravasation after inhibition of homotypic VE cadherin adhesion. These results provide support to the concept that VE-cadherin contributes to the normal differentiation of endothelial barrier function during CAM angiogenesis in vivo. PMID- 11037795 TI - The fibre dimensions of uterine smooth muscle of the rabbit following treatment by female sex steroids. AB - The effects of female sex hormones on the dimensions of myometrial smooth muscle fibres were studied by using ovariectomized rabbits. After one month of treatment, the fiber dimensions of the outer myometrial layer were measured, using cryostat sections. Calculated smooth muscle fiber volume was found to be in the sequence: control < medroxyprogesterone < estradiol < estradiol + medroxyprogesterone < estradiol alone. The measurements show that medroxyprogesterone-treated uteri contain the narrowest and the longest smooth muscle fibres, while estradiol treatment have the largest cells. This study complements previous observations in showing that medroxyprogesterone alone, or in combination with other modulators, contributes to sustain pregnancy by increasing internal resistance of estradiol-primed myometrial smooth muscle fiber fascicles. Our discussion, based on recent literature, shows that this resistance is ultimately controlled by changes in the myometrium innervation, in the repression of some controlling myofibrillar components, in the expression of specific membrane receptors and ionic channels, and in favoring the switching of molecular connexins in gap junctions, making P paramount in maintaining pregnancy. Moreover, other recent observations have also shown that probably an hcG-like hormone actually control P receptors expression in myometrial smooth muscles. PMID- 11037796 TI - Contractile and cytoskeletal proteins of smooth muscle cells in rat, rabbit, and human arteries. AB - The aim of this study was to determine whether similar populations of smooth muscle cells, in relation to contractile and cytoskeletal proteins, are present in normal and diseased human coronary arteries and normal and injured rat and rabbit arteries. Rat aortae and rabbit carotid arteries were de-endothelialised and the resulting neointimal thickening examined at set time points 2-24 weeks later. Immunohistochemistry revealed that arteries had three distinct populations of cells in respect to alpha-smooth muscle actin, smooth muscle myosin heavy chain and vimentin (staining intensities '-', '+' or '++' for each protein), but only two populations in respect to desmin ('-' and '+'). The different populations of cells were found in the neointima at all times after injury, in human atherosclerotic plaque and in the media of diseased, injured and uninjured vessels, although in different proportions. It was concluded that arteries of the human, rat and rabbit have cells with a wide spectrum of contractile and cytoskeletal proteins. Expression of the different proteins did not reflect the state of the artery after injury or during the disease process, and was not associated with the expansion of a subset of cells within the artery wall. PMID- 11037797 TI - Isolation and characterization of myogenic satellite cells from the muscular dystrophic hamster. AB - Myogenic satellite cells were isolated from control and dystrophic hamster diaphragms to examine cellular mechanisms involved in the physiology of muscular dystrophy. The Bio 14.6 dystrophic hamster, which possesses a defect in the delta sarcoglycan gene, develops biochemical and physical symptoms of Duchenne-like and limb girdle muscular dystrophies. Because primary cultures of the control and dystrophic satellite cells became extensively contaminated with non-myogenic cells during proliferation, cell clones were developed to provide pure cultures for study. Cell culture conditions were optimized with the use of Ham's F-12K medium containing 10% fetal bovine serum +5% horse serum + 10 ng/mL basic fibroblast growth factor + 50 microg/mL porcine gelatin. Proliferation rates of the two clonal cultures were similar between the two lines. Satellite cell derived myotubes from both primary cultures and clones differed between control and dystrophic animals. Dystrophic myotubes tended to be long and narrow, while the control-derived myotubes were broader. Measurement of muscle-specific creatine kinase during differentiation revealed that the dystrophic myotubes possessed higher creatine kinase levels than control myotubes (up to 146-fold at 168 h). The results demonstrate that satellite cells can be isolated from the hamster and may provide a useful tool to study muscular dystrophies associated with defects in the sarcoglycan complex and the involvement of sarcoglycans in normal skeletal muscle growth and development. PMID- 11037798 TI - Distribution of neurotransmitters and their effects on glucagon secretion from the in vitro normal and diabetic pancreatic tissues. AB - The distribution of adrenergic, cholinergic and amino acid neurotransmitters and/or their enzymes were examined in both the normal and diabetic pancreatic tissues in rat using immunohistochemistry to determine whether changes in the pattern of distribution of nerves containing these neurotransmitters will occur as a result of diabetes mellitus. In addition to this, the effect of noradrenaline (NA), adrenaline (ADR), acetylcholine (ACh) and gamma-amino butyric acid (GABA) on glucagon secretion from the isolated normal and diabetic pancreatic tissues was also investigated. Pancreatic fragments from the tail end of normal and diabetic rats were removed and incubated with different concentrations (10(-8)-10(-4) M) of these neurotransmitters. Glucagon secretion into the supernatant was later determined by radioimmunoassay. NA at 10(-6) M evoked a three-fold increase in glucagon secretion from normal pancreatic tissue fragments. In diabetic pancreatic tissue, NA at 10(-6) M was able to increase glucagon secretion 1.5 times the value obtained from diabetic basal. ADR (10(-8) M) increased glucagon secretion slightly but not significantly in normal pancreatic tissue. ADR inhibited glucagon secretion from diabetic pancreas at all concentrations. ACh (10(-8) M) induced a five-fold increase in glucagon secretion from normal pancreatic tissue. In a similar way, ACh evoked a two-fold increase in glucagon secretion from diabetic pancreas at 10(-4) M. In normal pancreatic tissue, GABA produced a slight but not significant increase in glucagon secretion at 10(-4) M. In contrast to this it inhibited glucagon secretion from diabetic pancreatic tissue fragments at all concentrations. In summary, tyrosine hydroxylase- and choline acetyltransferase-positive nerves are equally well distributed in both normal and diabetic rat pancreas. There was an increase in the number of glucagon positive cells and a decrease in the number of GABA positive cells in diabetic pancreas. NA and ACh have a potent stimulatory effect on glucagon secretion from normal pancreatic tissue fragments, whereas ADR and GABA produced a small but not significant increase in glucagon secretion from normal pancreas. NA and GABA stimulated glucagon secretion from diabetic pancreas. In contrast, ADR and ACh inhibited glucagon secretion from diabetic pancreas. Neurotransmitters vary in their ability to provoke glucagon secretion from either normal or diabetic pancreas. PMID- 11037799 TI - Elucidation of the genetic basis of the common 'intermediate metabolizer' phenotype for drug oxidation by CYP2D6. AB - A subgroup of 10-15% of Caucasians are termed phenotypical 'intermediate metabolizers' of drug substrates of CYP2D6 because they have severely impaired yet residual in-vivo function of this cytochrome P450. Genotyping based on the currently known CYP2D6 alleles does not predict this phenotype satisfactorily. A systematic sequencing strategy through 1.6 kb of the CYP2D6 5'-flanking sequence revealed six mutations of which three were exclusively associated with the functional CYP2D6*2 allele (-1496 C to G; -652 C to T; and -590 G to A), two were associated with the nonfunctional *4 and with the functional *10-alleles (-1338 C to T and -912 G to A) and one (-1147 A to G) was seen in all *2, *4 and *10 alleles investigated. The -1496 C to G mutation was found to be polymorphic within CYP2D6*2 alleles. In a family study, the wild-type CYP2D6 *2[-1496 C] and the novel variant [-1496 G] allele co-segregated with lower and higher CYP2D6 in vivo function, respectively, as shown by phenotyping using sparteine as probe drug. In a representative population sample selected for genotypes comprising one CYP2D6*2 and one non-functional allele, the median urinary metabolic ratio (MRs) for sparteine oxidation was 4.4-fold reduced in individuals with the variant allele (*2[-1496 G], MRs = 0.53, n = 27) compared with individuals lacking the mutation (*2[-1496 C], MRs = 2.33, n = 12; P < 0.0001). The mutation -1496 C to G has an estimated frequency of approximately 20% in the general population and allows establishment of a genotype for the identification of over 60% of intermediate metabolizers in Caucasian populations. PMID- 11037801 TI - Role of cytochrome P450 2D6 (CYP2D6) in the stereospecific metabolism of E- and Z doxepin. AB - The tricyclic antidepressant, doxepin, is formulated as an irrational mixture of E (trans) and Z (cis) stereoisomers (85%: 15%). We examined the stereoselective metabolism of doxepin in vitro, with the use of human liver microsomes, recombinant CYP2D6 and gas chromatography-mass spectrometry. In human liver microsomes over the concentration range 5-1500 microM, the rate of Z-doxepin N demethylation exceeded that of E-doxepin above 100 microM in two of three livers. Eadie-Hofstee plots were curvilinear indicating the involvement of several enzymes in N-demethylation. Coincubation of doxepin with 7,8-naphthoflavone and ketoconazole reduced the rates of N-demethylation of E- and Z-doxepin by 30-50% and 40-60%, respectively, suggesting the involvement of CYP1A and CYP3A4, whilst quinidine had little effect on N-demethylation. In contrast, doxepin hydroxylation was exclusively stereo-specific; E-doxepin and E-N-desmethyldoxepin were hydroxylated with high affinity in liver microsomes and by recombinant CYP2D6 (Km in the range of 5-8 microM), but there was no evidence of Z-doxepin hydroxylation. In 'metabolic consumption' experiments with liver microsomes (having measurable CYP2D6 activity) and initial substrate concentration of 1 microM, the consumption of E-doxepin was greater (P < 0.05, n = 5) than that of Z doxepin. Quinidine inhibited the consumption of E-doxepin but did not affect the consumption of Z-doxepin. With N-desmethyldoxepin, quinidine inhibited the consumption of E-N-desmethyl-doxepin whereas Z-N-desmethyldoxepin appeared to be a terminal oxidative metabolite. In summary, CYP2D6 is a major oxidative enzyme in doxepin metabolism; predominantly catalysing hydroxylation with an exclusive preference for the E-isomers. The relatively more rapid metabolism of E-isomeric forms, and the limited metabolic pathways for the Z-isomers may explain the apparent enrichment of Z-N-desmethyldoxepin that is observed in vivo. PMID- 11037800 TI - Prospective CYP2D6 genotyping as an exclusion criterion for enrollment of a phase III clinical trial. AB - A phase III study was performed to compare the efficacy and safety of lamotrigine (Lamictal), desipramine (Norpramin), and placebo in the treatment of unipolar depression. Desipramine is extensively metabolized by cytochrome P450 2D6 (CYP2D6), and kinetics of this compound are altered in poor metabolizers. Genotyping was utilized to exclude poor metabolizers in order to increase subject safety and to eliminate the need to continuously monitor plasma desipramine levels. As part of screening, subjects were genotyped for the *3(A), *4(B), and *5(D) alleles, which identify approximately 95% of poor metabolizers. Extensive metabolizers were eligible for randomization to the lamotrigine, desipramine, or placebo arm. Follow-up genotyping for the *6(T) and *7(E) alleles was performed after study enrollment and was used to identify poor metabolizers who may have been incorrectly identified as extensive metabolizers upon initial three-allele screening. Of 628 subjects screened for *3(A), *4(B), *5(D) alleles, 590 (93.9%) were classified as extensive metabolizers. The remaining 38 (6.1%) subjects were poor metabolizers and excluded. Subsequent *6(T) and *7(E) testing revealed that two poor metabolizers had been enrolled, and the follow-up genotyping provided an explanation for the high desipramine plasma concentrations in one subject. No differences in phenotypic or allelic frequencies were found between the study population and literature populations. However, the frequency of poor metabolizers varied among clinical sites (0-15%). For a compound that is extensively metabolized by CYP2D6, prescreening subjects for *3(A), *4(B), *5(D), *6(T) and *7(E) alleles can increase subject safety and eliminate the need to continuously monitor drug plasma concentrations. PMID- 11037802 TI - Poor metabolizers at the cytochrome P450 2D6 and 2C19 loci are at increased risk of developing adult acute leukaemia. AB - We have genotyped over 550 cases of acute leukaemia and 950 matched controls from a population-based case-control study, to investigate the impact cytochrome P450s 2D6, 2C19 and 1A1 have on susceptibility to adult acute leukaemia. Analysis included potential associations between polymorphic status and acute myeloid leukaemia (AML), acute lymphoblastic leukaemia (ALL), plus the FAB and cytogenetic subtypes therein. A significant increased risk was found for CYP2D6 poor metabolizer phenotype and acute leukaemia [odds ratio (OR) = 1.69, 95% confidence interval (CI) 1.17-2.43], a risk also found in AML and ALL. No interaction was found with smoking. However, a significant age-related association between CYP2D6 polymorphism and acute myeloid leukaemia implied that the excess risk was confined to persons aged 40 years and over (OR 2.38, 95% CI 1.53-3.71). Amongst AML cases, increased odds ratios were observed in both de novo (OR 1.54, 95% CI 1.02-2.32) and secondary leukaemia (OR 2.83, 95% CI 0.91 8.77), and among patients with a chromosomal abnormality (OR 2.00, 95% CI 1.11 3.61). An increased risk was found for the CYP2C19 poor metabolizer phenotype (OR 1.68, 95% CI 0.97-2.92) which was also present in AML and ALL. For this CYP450 locus, an increased risk was suggested in secondary leukaemia (OR 2.67, 95% CI 0.44-16.3) and amongst AML cases with a chromosomal abnormality (OR 6.72, 95% CI 2.22-20.4). No difference in CYP1A1 genotype distribution was found for acute leukaemia, AML, ALL or any other diagnostic classification group used. No significant interactions between CYP2D6, CYP2C19 or CYP1A1 were found. PMID- 11037803 TI - Relation between inducibility of CYP1A1, GSTM1 and lung cancer in a French population. AB - Smoking is the principal cause of lung cancer. However, not all smokers will develop this disease. Individual susceptibility to chemically induced cancer may be explained in part by genetic differences in the activation and detoxification of procarcinogens. The activation phase of polycyclic aromatic hydrocarbon (PAH) metabolism is governed by the enzyme CYP1A1, induced by PAH when it enters the body. The extent to which PAH induces CYP1A1 activity varies greatly from one subject to another. CYP1A1 inducibility has long been associated, although inconsistently, with an increased risk of lung cancer. In 1982, Kouri corroborated Kellerman's results with a new method for measuring inducibility, but few studies have reported using this method. The glutathione S-transferases (GSTs) are involved in the detoxification phase of PAH, and the allelic deletion of GSTM1 has been also associated with an increased risk of lung cancer. We conducted a case-control study to examine the risk of lung cancer related, separately and together, to CYP1A1 inducibility, GSTM1 polymorphism and cigarette smoking in a French population. The 611 subjects were 310 incident lung cancer cases and 301 hospital control subjects. We were able to constitute a DNA bank for 552 subjects (89.5%) and gather detailed information on smoking history for all of them. Inducibility could be measured for 195 cases and 183 control subjects. Results for GSTM1 polymorphism concern 247 cases and 254 control subjects. GSTM1 polymorphism and inducibility could both be assessed for 179 cases and 166 control subjects. The odds ratio related to inducibility was 1.7 [1.0-3.0] for medium and 3.1 (1.3-7.4) for hyper inducers. The association with GSTM1 was 1.6 (1.0-2.6). With a reference category of subjects who were both low inducers and GSTM1(+), we found an odds ratio for lung cancer of 8.1 (2-31) for the subjects with both risk factors [i.e. GSTM1(-) and hyper inducers]. Our data did not reveal evidence of interaction between smoking and inducibility. On the other hand, we found an interaction of 3.6 (0.6-21) between inducibility and GSTM1. PMID- 11037804 TI - Structural heterogeneity at the UDP-glucuronosyltransferase 1 locus: functional consequences of three novel missense mutations in the human UGT1A7 gene. AB - One of the most important mechanisms involved in host defense against xenobiotic chemicals and endogenous toxins is the glucuronidation catalysed by UDP glucuronosyltransferase enzymes (UGT). The role of genetic factors in determining variable rates of glucuronidation is not well understood, but phenotypic evidence in support of such variation has been reported. In the present study, six single nucleotide polymorphisms were discovered in the first exon of the UGT1A7 gene, which codes for the putative substrate-binding domain, revealing a high structural heterogeneity at the UGT1 gene locus. The new UGT1A7 proteins differ in their primary structure at amino acid positions 129, 131 and 208, creating four distinct UGT1A7 allelic variants in the human population: UGT1A7*1 (N129 R131 W208), *2 (K129 K131 W208), *3 (K129 K131 R208), and *4 (N129 R131 R208). In functional studies, HEK cells stably transfected to express the four allelic UGT1A7 variants exhibited significant differences in catalytic activity towards 3 , 7-, and 9-hydroxy-benzo(a)pyrene. UGT1A7*3 exhibited a 5.8-fold lower relative Vmax compared to wild-type *1, whereas *2 and *4 had a 2.6- and 2.8-fold lower relative Vmax than *1, respectively, suggesting that these mutations confer slow glucuronidation phenotype. Kinetic characterization suggested that these differences were primarily attributable to altered Vmax. Additionally, it suggested that each amino acid substitutions can independently affect the UGT1A7 catalytic activity, and that their effects are additive. The expression pattern of UGT1A7 studied herein and its catalytic activity profile suggest a possible role of UGT1A7 in the detoxification and elimination of carcinogenic products in lung. A population study demonstrated that a considerable proportion of the population (15.3%) was found homozygous for the low activity allele containing all three missense mutations, UGT1A7*3. These findings suggest that further studies are needed to investigate the impact of the low UGT1A7 conjugator genotype on individual susceptibility to chemical-induced diseases and responses to therapeutic drugs. PMID- 11037805 TI - Glutathione transferase T1 phenotype affects the toxicokinetics of inhaled methyl chloride in human volunteers. AB - The aim of the present study was to investigate how the genetic polymorphism in glutathione transferase T1 (GSTT1) affects the metabolism and disposition of methyl chloride in humans in vivo. The 24 volunteers (13 males and 11 females) who participated in the study were recruited from a group of 208 individuals previously phenotyped for GSTT1 by measuring the glutathione transferase activity with methyl chloride in lysed erythrocytes ex vivo. Eight individuals with high (+/+), eight with medium (+/0) and eight with no (0/0) GSTT1 activity were exposed to methyl chloride gas (10 p.p.m.) in an exposure chamber for 2 h. Uptake and disposition was studied by measuring the concentration of methyl chloride in inhaled air, exhaled air and blood. A two-compartment model with two elimination pathways corresponding to exhalation and metabolism was fitted to experimental data. The average net respiratory uptake of methyl chloride was 243, 158, and 44 micromol in individuals with high, intermediate and no GSTT1 activity, respectively. Metabolic clearance was high (4.6 l/min) in the +/+ group, intermediate (2.4 l/min) in the +/0 group, and close to zero in 0/0 individuals, while the exhalation clearance was similar in the three groups. No exposure related increase in urinary S-methyl cysteine was detected. However, gender and the GSTTl phenotype seemed to affect the background levels. In conclusion, GSTT1 appears to be the sole determinant of methyl chloride metabolism in humans. Thus, individuals with nonfunctional GSTT1 entirely lack the capacity to metabolize methyl chloride. PMID- 11037806 TI - Pharmacological properties of the naturally occurring Phe-124-Cys variant of the human 5-HT1B receptor: changes in ligand binding, G-protein coupling and second messenger formation. AB - The aim of this study was to analyse whether substitution of phenylalanine in position 124 of the human (h) 5-HT1B receptor by cysteine, a naturally occurring variant of this receptor, modifies not only ligand binding, but also G-protein coupling and second messenger formation. Stably transfected rat C6 glioma cells, which express either the h5-HT1B variant receptor (VR) or the wild-type receptor (WTR) were used. In saturation experiments with [3H]5-carboxamidotryptamine ([3H]5-CT), the maximum binding (Bmax) of the VR amounted to only 60% of that to WTR. In competition experiments with 1 nM [3H]5-CT, the following 5-HT receptor ligands exhibited a higher affinity for the mutant receptor than for the WTR: L 694,247, 5-CT, 5-HT, sumatriptan (agonists listed at decreasing order of potency) and SB-224289 (a selective h5-HT1B receptor inverse agonist with competitive antagonistic properties). In contrast, the mixed 5-HT1B/1D receptor antagonist GR 127935 exhibited equal affinity for both isoforms. The efficacy of L-694,247, 5 CT, 5-HT and sumatriptan in stimulating [35S]GTPgammaS binding (a measure of G protein coupling) to membranes of cells expressing the VR was approximately 50 65% lower compared to membranes of cells expressing the WTR, but their potency was 2.8-3.6-fold higher. SB-224289, which decreased [35S]GTPgammaS binding when given alone, but not GR-127935, was more potent in antagonizing the stimulatory effect of 5-CT on [35S]GTPgammaS binding to membranes expressing the VR compared to membranes expressing the WTR. In whole cells expressing the VR, 5-CT and sumatriptan inhibited the forskolin-stimulated cAMP accumulation 3.2-fold more potently than in cells expressing the WTR. In conclusion, our data suggest that the Phe-124-Cys mutation modifies the pharmacological properties of the h5-HT1B receptor and may account for pharmacogenetic differences in the action of h5-HT1B receptor ligands. Thus, the sumatriptan-induced vasospasm which occurs at low incidence as a side-effect in migraine therapy may be related to the expression of the (124-Cys)h5-HT1B receptor in patients with additional pathogenetic factors such as coronary heart disease. PMID- 11037807 TI - Factors in the pathogenesis of tumors of the sphenoid and maxillary sinuses: a comparative study. AB - OBJECTIVES/HYPOTHESIS: To explain the processes that lead to the development of tumors in the maxillary and sphenoid sinuses. STUDY DESIGN: A 32-year review of the world's literature on neoplasms of these two sinuses and a randomized case controlled study comparing the normal mucosal architecture of the maxillary to the sphenoid sinus. METHODS: Analysis of a 32-year world literature review reporting series of cases of maxillary and sphenoid sinus tumors. Tumors were classified by histological type and separated into subgroups if an individual incidence rate was reported. Histomorphometry of normal maxillary and sphenoid sinus mucosa was performed in 14 randomly selected patients (10 sphenoid and 4 maxillary specimens). Specimens were fixed in 10% formalin, embedded in paraffin, and stained with periodic acid-Schiff (PAS) and hematoxylin. Histomorphometric analysis was performed with a Zeiss Axioscope light microscope (Carl Zeiss Inc., Thornwood, NY) mounted with a Hamamatsu (Hamamatsu Photonics, Tokyo, Japan) color chilled 3 charge coupled device digital camera. The images were captured on a 17 inch Sony (Sony Corp., Tokyo, Japan) multiscan monitor and analyzed with a Samba 4000 Image Analysis Program (Samba Corp., Los Angeles, CA). Five random areas were selected from strips of epithelium removed from each sinus, and goblet and basal cell measurements were made at magnifications x 100 and x 400. RESULTS: The literature review revealed that the number and variety of tumors in the maxillary sinus are much greater than those in the sphenoid. The incidence of metastatic lesions to each sinus is approximately equal. No recognized pattern of spread from any particular organ system could be determined. On histomorphometric study there were no statistically significant differences between the sinuses in the concentration of goblet cells, basal cells, or seromucinous glands. CONCLUSIONS: Factors involved in the pathogenesis of tumors of the maxillary and sphenoid sinuses include differences in nasal physiology, embryology, morphology, and topography. There are no significant histological differences in the epithelium and submucous glands between the two sinuses to explain the dissimilar formation of neoplasms. PMID- 11037808 TI - Otolaryngology 1900-1999: a century of progress? PMID- 11037809 TI - Building the powerful 10-minute office visit, Part I: Introduction to the new section. AB - OBJECTIVE: This is the first of a planned progressive series of articles entitled "Tutorials in Clinical Research" written to assist those who would become more active in the application of scientific methods to their practices and to assist those who would seek to develop and publish new discoveries of their own. On completion of Tutorials, the reader should be able to 1) critically and efficiently read the literature and 2) use his or her personal practice as a laboratory for clinical investigation. STUDY DESIGN: Tutorial. METHODS: The Journal editor agreed to launch the series "Tutorials in Clinical Research," which was conceived out of a need for an easily understandable and accessible collection of tutorials for the busy practitioner. The Clinical Research Working Group at Washington University, Department of Otolaryngology-Head and Neck Surgery, will write, edit, and solicit additional authors for the completion of this planned 22-article series. The intended audience includes, among other groups, practitioners without research funds who are interested in the application of scientific methods to their practices. RESULTS: This first article in the series outlines how to begin and three steps for the initial application of scientific methods to practice, including critical literature review, prospective analysis of practice, and specific project development. CONCLUSIONS: The application of scientific methods to practice can be fun, educational, and effective in the enhancement of quality care. The "value added" result is the development of the powerful 10-minute patient office visit for the busy practitioner. PMID- 11037810 TI - Workforce issues for the academic otolaryngologist. AB - BACKGROUND: The current medico-economic environment has led to profound changes in our health care system and questions of physician surplus. These issues have particularly affected the academic health care system, as research funding and departmental support have decreased, and many young otolaryngologists are questioning academic careers because of these uncertainties. The current study was undertaken to assess the workforce environment for the academic otolaryngologist, particularly the young physician. METHODS: Surveys were sent to the academic chairmen of all accredited otolaryngology residency programs in the United States, requesting information on faculty appointments--actual and projected-as well as subspecialty appointments and expectations of young faculty. RESULTS: The response rate was 60% (59/98). Faculty additions have been relatively stable from 1994 to 1998, with approximately 37 assistant professor and 5 associate professor positions filled yearly. Faculty additions were the result of departmental expansion in 83% of cases and spanned many subspecialties. The subspecialty positions most frequently added from 1994 to 1998 were generalists (57), head and neck oncologists (53), pediatric otolaryngologists (48), and otologists (39), with generalists filling 15 positions in 1998. Ninety three percent of programs anticipate faculty additions in the next 5 years; most will be at the assistant professor level (77%), with 30% of positions for generalists, 20% for head and neck oncologists, and 18% for pediatric otolaryngologists. Faculty expectations are primarily clinical, with research being least important. CONCLUSIONS: Academic positions are available for the young otolaryngologist, particularly in the fields of general otolaryngology, head and neck oncology, and pediatric otolaryngology. PMID- 11037811 TI - Efficacy of transoral intraluminal Wallstents for tracheal stenosis or tracheomalacia. AB - OBJECTIVES: The efficacy and safety of intraluminal Wallstent Endoprosthesis (Boston Scientific/ Medi-Tech, Quincy, MA) placement to restore airway patency in patients with tracheal stenosis or tracheomalacia are unknown. STUDY DESIGN: Retrospective review in setting of tertiary, referral, and academic center. METHODS: A retrospective review of 13 consecutive patients over a 2-year period who underwent transoral resection of tracheal stenosis and immediate transoral Wallstent placement. One patient had tracheomalacia. All of the patients were considered at high risk for transcervical surgery or had failed prior traditional open procedures. RESULTS: The average patient age was 54.2 years, with nine male and four female patients. All had Cotton/Myer stenoses (grades II to IV) with moderate to severe degrees of inspiratory stridor. Four patients were tracheotomy dependent. The length of stenosis varied from 1 to 4 cm. One patient had a 10-cm segment of tracheomalacia. At the time of writing, none of the patients has had a problem with significant migration or extrusion and most of the patients have incorporated the stent well without any short-term obstructive granulation tissue. After a mean follow-up of 15 months (range, 4-24 mo). 10 of the 12 patients with stenosis (83%) have remained free of any inspiratory noise during breathing. The one patient with tracheomalacia also has remained free of symptoms. CONCLUSIONS: Transoral Wallstents appear to be safe and may be a reasonable alternative in the restoration of airway patency in select patients with tracheal stenosis or tracheomalacia. PMID- 11037812 TI - Endoscopic sinus surgery in patients infected with HIV. AB - OBJECTIVES: To be able to plan appropriate surgical treatment for patients with HIV infection who have sinusitis refractory to medical therapy. DESIGN: We retrospectively reviewed the charts of 186 patients with HIV who required surgical treatment for sinusitis between 1987 and 1998. One hundred six charts provided the necessary information and an adequate follow-up to be included in the study. Collected data included preoperative and postoperative symptoms, radiographic staging, CD4 count at the time of surgery when available, and type and extent of surgery. RESULTS: Surgical treatment evolved over the 12 years from limited surgery to standard endoscopic sinus surgery (ESS). Eighteen patients had invasive fungal disease or complications of sinusitis requiring radical surgery. Thirty-six patients were treated with minimal procedures to address involved sinuses only. These patients were treated between 1987 and 1991. Recurrent disease or further complications occurred in 80.6% of the patients in this group. Since 1992, 52 patients were treated with standard ESS following the same indications for HIV- patients. This group had an improvement of symptoms in 75% of the cases, a rate comparable to the success rate in HIV- patients. CONCLUSIONS: HIV+ patients undergoing standard ESS enjoy a satisfactory success rate. HIV+ patients with surgical indication for endoscopic sinus surgery should be treated as non-HIV+ patients. Apparently, low CD4 count (< 100) does not serve as a contraindication for definitive surgery. PMID- 11037813 TI - A clinicopathological study of sinonasal neuroendocrine carcinoma and sinonasal undifferentiated carcinoma. AB - OBJECTIVE: Sinonasal undifferentiated carcinoma (SNUC) and sinonasal neuroendocrine carcinoma (SNEC) are relatively newly recognized, rare entities requiring further clinicopathological analysis to advance our understanding and determine prognostic distinctions between them. STUDY DESIGN: Retrospective chart review. METHODS: Cases were retrieved from the Copath system. One patient was seen in consultation from an outside institution. Histological and immunohistochemical findings, patient demographics, treatment regimens, and outcomes were analyzed and compared. RESULTS: Ten patients (7 men, 3 women) ranging in age from 17 to 58 years (mean age, 44.7 y) were included. Four patients were classified with SNEC, six as having SNUC. The predominant site was the superior nasal cavity or ethmoids (seven cases), followed by the maxilla (four cases). Disease in four patients was clinically staged as N1 (three with SNUC, one with SNEC), and in six patients as NO (three with SNEC, three with SNUC). Of the nine patients who were treated initially with surgical resection, seven received postoperative radiation therapy alone, one received postoperative radiation and chemotherapy, and one had only limited postoperative chemotherapy. One patient was treated with radiation therapy and chemotherapy alone, without surgical resection. Follow-up was obtained ranging from 6 to 108 months (mean period, 26.4 mo). Three patients died of disease 10, 14, and 41 months, respectively, after diagnosis. Three patients had persistent disease at 6, 9, and 21 months, respectively, two of them with distant metastases. Four patients were disease free after 6, 18, 31, and 108 months, respectively. CONCLUSIONS: SNUC and SNEC are both aggressive tumors, usually presenting in middle age as a nasal mass. Both tumors have the capacity to metastasize locally and distantly, and both can result in poor outcomes. This small series precludes a demographic or prognostic distinction between the two groups. PMID- 11037814 TI - More aggressive behavior of squamous cell carcinoma of the anterior tongue in young women. AB - OBJECTIVE: To review the combined experience from two large medical centers in treating young female patients with anterior tongue cancer to determine the clinical course of this unique subset of patients. STUDY DESIGN: Retrospective study. METHODS: Seventeen female patients less than 40 years of age (group A) and 17 older patients, both male and female, greater than 40 years of age (group B) who had treatment for invasive squamous cell carcinoma of the anterior tongue were studied. The charts were reviewed for the clinical staging, treatment, and outcome of each patient. The disease-free survival and recurrence rates were compared between the two cohorts. RESULTS: The mean disease stage between the groups was II. The survival analysis showed a significant difference between the two groups in recurrences (group A = 65%, group B = 41%; P = .02). Further, of the patients who had recurrence, the young women did so significantly earlier in their disease course than the older patients (group A = 14 mo, group B = 40 mo; P < .05). Although the survival differences did not reach statistical significance (P = .15), the power of the study was low (power = 0.26) resulting in a high level type II error. CONCLUSION: These data suggest that young women with squamous cell carcinoma of the anterior tongue have significantly higher rates of recurrent disease and the interval to recurrence is significantly shorter than in older patients. Further investigation is warranted until a statistically significant cohort is accrued; until that time, these patients warrant an aggressive initial treatment and close surveillance for recurrence. PMID- 11037815 TI - Prudent management of the mid-cheek mass: revisiting the accessory parotid gland tumor. AB - OBJECTIVES/HYPOTHESIS: The head and neck surgeon's fascination with parotid surgery arises from the gland's spectrum of histopathological presentations, as well as the diversity of its morphological features. A mass arising in the mid cheek region may often be overlooked as a rare accessory lobe parotid neoplasm. This report serves to revisit the topic of accessory parotid gland neoplasms to emphasize proper management, particularly the surgical aspects, so that consequences of salivary fistula, facial nerve paralysis, and recurrence are avoided. STUDY DESIGN: This is a retrospective review of our experience with four accessory parotid gland neoplasms and five other masses mimicking this lesion. METHODS: A literature review and retrospective chart review. RESULTS: Over a 6 year period, we have encountered four true accessory lobe tumors, all pleomorphic adenomas. These presented very similarly to four other more commonly encountered masses not of salivary origin and one normal but hyperplastic accessory parotid gland. All were removed through a wide parotidectomy-style approach modified by extending incisions anterosuperiorly and inferoanteriorly. The only complication was a minor salivary fistula in one patient. There were no permanent facial paralyses. CONCLUSIONS: Accessory parotid gland neoplasms are rare and may present as innocuous extraparotid mid-cheek masses. A high index of suspicion, prudent diagnostic skills (including fine-needle aspiration [FNA] biopsy followed by computed tomography [CT] imaging), and meticulous surgical approach (extended parotidectomy-style incision and limited peripheral nerve dissection when possible) are the keys to successful management of these lesions. PMID- 11037816 TI - External beam radiation followed by planned neck dissection and brachytherapy for base of tongue squamous cell carcinoma. AB - BACKGROUND: Surgical resection of tongue base cancer can leave the patient with significant functional deficits. Other therapies, such as external beam radiation followed by neck dissection and radiation implants, have shown equal tumor control with good functional outcome. METHODS: Between March 1991 and July 1999, 12 patients at Oregon Health Sciences University, the Portland Veterans Administration Medical Center and West Virginia University School of Medicine Hospital were treated with external beam radiation followed by neck dissection and Ir192 implants. Two patients had T1 disease, two had T2, five patients had T3 tumors, and three had T4 tumors. Six had N2a necks, three had N2b necks, and three had N2c. Follow-up ranged from 13 months to 8 years. RESULTS: After external beam radiation, five patients had complete response and seven had partial response in the neck without complications. One patient underwent a unilateral radical neck dissection, eight had unilateral selective neck dissections involving levels I to IV, and three had dissections involving levels I to III. One of the five patients who had a complete clinical response in the neck had pathologically positive nodes. One patient had a pulmonary embolus that was treated and had no permanent sequelae. There were three complications from brachytherapy. Two patients had soft tissue necrosis at the primary site and one patient had radionecrosis of the mandible. All healed without further therapy. One patient had persistent disease and underwent a partial glossectomy but died of local disease. Distant metastasis developed in two patients. All others show no evidence of disease and are able to eat a normal diet by mouth. CONCLUSION: This combination of therapies should be considered when treating tongue base cancer. PMID- 11037817 TI - Incidence of unsuspected metastases in lateral cervical cysts. AB - OBJECTIVE: Solitary cystic squamous cell carcinoma metastases may be difficult to distinguish clinically from a benign cervical cyst. We sought to identify the incidence of solitary cystic squamous cell carcinoma metastasis in patients presenting with apparently benign cervical cysts. STUDY DESIGN: Retrospective review. METHODS: The records of all patients who presented with isolated lateral cervical cysts between 1983 and 1999 were reviewed. Patients with a clinically apparent primary malignancy, a history of head and neck cancer, a history of irradiation, or age less than 18 years were excluded from analysis, as were patients with a histological diagnosis of nonsquamous cell malignancy or those without a final histological diagnosis. RESULTS: One hundred twenty-one adult patients presented with an initial diagnosis of lateral cervical cyst. Metastatic squamous cell carcinoma was demonstrated histologically after surgical excision in 12 patients (9.9%). The incidence of malignancy was significantly greater in patients greater than 40 years of age (23.5%, P < .0001). Results of preoperative fine-needle aspiration (FNA) were negative for malignancy in five cases of metastatic squamous cell carcinoma. Panendoscopy with directed biopsies revealed an occult primary in the base of tongue in three patients, tonsil in one patient, and nasopharynx in one. No primary was found in six patients, despite repeated examinations and close follow-up. CONCLUSIONS: Solitary cervical cysts in patients older than 40 years of age should be presumed to be carcinoma until proven otherwise. A negative FNA result may be misleading, because of hypocellularity of the cyst fluid. Excisional biopsy should be undertaken with provisions made for frozen-section analysis of the specimen and contingency panendoscopy with directed biopsies of Waldeyer's ring if frozen-section histological examination reveals malignancy. PMID- 11037818 TI - Glutathione S-transferase pi in squamous cell carcinoma of the head and neck. AB - OBJECTIVES/HYPOTHESIS: Oxidative/reductive (redox) DNA damage from radical species such as nitric oxide (NO*) are increasingly being implicated in the development of cancer. Moreover, redox-protective cellular mechanisms, such as glutathione S-transferase, may determine cellular susceptibility to this redox mediated damage. METHODS: Formalin-fixed, paraffin-embedded tissue samples of 11 normal oral mucosa, 15 reactive/dysplastic lesions, and 131 head and neck squamous cell carcinomas (HNSCCs) were immunohistochemically stained using a polyclonal antibody against glutathione S-transferase pi (GST-pi). Slides were reviewed in a blinded fashion by the study pathologist (G.K.H.) and intensity was graded, noting the pattern of immunostaining. These staining characteristics were compared with those obtained using monoclonal antibodies against endothelial constitutive nitric oxide synthase (ecNOS) and nitrotyrosine, a marker of NO*'s pathological nitrosylation of proteins on serial sections of the same tissue. Patient charts were reviewed and clinical data collected. RESULTS: The expression of GST-pi was significantly increased in reactive/dysplastic and HNSCC samples compared with normal squamous mucosa (P < .001 for both). Furthermore, among the carcinomas the expression of GST-pi was significantly increased in higher-grade lesions (P < .02). The expression of GST-pi correlated highly with the expression of ecNOS and nitrotyrosine (P < .0001 for both). CONCLUSIONS: These findings demonstrate that GST-pi is upregulated in conjunction with the NO*-generating ecNOS isoform and implicate GST-pi in protecting HNSCC from the cytotoxic effects of high concentrations of NO* found in the tumor bed. PMID- 11037819 TI - Therapeutic efficacy of Th1 and Th2 L-selectin--CD4+ tumor-reactive T cells. AB - OBJECTIVE: To evaluate the cytokine secretion profile and therapeutic efficacy of Th1 CD4+ L-selectin-tumor-draining lymph node lymphocytes in the treatment of murine pulmonary metastases. STUDY DESIGN: Prospective, murine in vivo and in vitro study. METHODS: B6 mice were injected bilaterally subcutaneously with MCA 205 sarcoma cells to initiate tumor growth. Eleven days later, tumor-draining inguinal lymph nodes were harvested. Single-cell suspensions were prepared and fractionated using magnetically activated cell sorting. Sorted CD4+ L-selectin lymphocytes were activated with anti-CD3 monoclonal antibody for 48 hours either alone to give a Th1 phenotype, or in the presence of interleukin (IL)-4 and anti interferon-gamma (alpha-IFN-gamma) monoclonal antibody to elicit a Th2 phenotype. Activated cells were then expanded for 3 days in IL-2. Resulting cells were used to treat 3-day pulmonary metastases. Enzyme-linked immunosorbent assay and intracellular fluorescent-activated cell-sorter (FACS) scanning were used to evaluate the cytokine secretion profiles of these cells. RESULTS: Activated and expanded L-selectin- CD4+ T cells demonstrated a Th1 cytokine profile and excellent antitumor efficacy. In contrast, L-selectin- CD4+ lymphocytes activated in the presence of IL-4 and alpha-IFN-gamma monoclonal antibody demonstrated a Th2-like profile and significantly (P < .05) poorer antitumor efficacy. CONCLUSIONS: The cytokine environment during the activation of tumor-draining lymph nodes can influence the therapeutic efficacy of activated L-selectin-, CD4+ T cells. Cell mediated, Th1-dependent immunity appears to play an important role in mediating tumor regression. Culture conditions promoting Th2 cells resulted in T cells associated with diminished antitumor efficacy. PMID- 11037820 TI - Elimination of temporal bone cerebrospinal fluid otorrhea using hydroxyapatite cement. AB - OBJECTIVE: This report introduces a new method to control cerebrospinal fluid (CSF) otorrhea using hydroxyapatite cement (HAC) via a transmastoid approach. This technique eliminates the need for a transmastoid or middle cranial fossa approach with soft tissue repair and prolonged hospitalization caused by lumbar drainage. STUDY DESIGN: Retrospective review. METHODS: Thirteen cases of transmastoid repairs of CSF otorrhea using HAC from August 1996 to February 1999 were reviewed. RESULTS: The CSF leak was controlled in every patient using HAC through a transmastoid approach. The reconstruction involved eight tegmen defects, three posterior fossa dural plate defects, and two congenital inner ear fistula secondary to Mondini malformation. Postoperative wound infection in one patient was the only complication that occurred. The average hospital stay was 48 hours. Follow-up ranged from 12 to 44 months with no recurrence of CSF otorrhea. CONCLUSION: The successful use of HAC to control CSF otorrhea through a transmastoid approach reduces patient morbidity by obviating the need for middle cranial fossa approaches, donor soft tissue sites, and spinal drainage. PMID- 11037821 TI - Selective indications for the management of extensive anterior epitympanic cholesteatoma via combined transmastoid/middle fossa approach. AB - OBJECTIVES/HYPOTHESIS: Cholesteatoma that is present in the anterior epitympanic space may extend medially along the supralabyrinthine route to the geniculate ganglion, labyrinth, and cochlea and medially toward Kawase's triangle and the anterior petrous apex. Superiorly it may erode into the middle fossa. Contemporary microsurgical techniques allow for optimal management of these lesions with minimal morbidity, provided that the irregular and complex osteology of the petrous base is understood. The objective of the study was to review the relevant regional anatomy, pathobiology, and current algorithm used in treatment of this select patient population using a combined transmastoid/middle fossa (TM/MF) approach. METHODS: A retrospective review was performed of all clinical and radiographic data from patients undergoing combined TM/MF management of extensive anterior epitympanic cholesteatoma between July 1984 and June 1998. Data from physical examinations, preoperative imaging studies, and operative findings and other relevant data were tabulated and analyzed for patients undergoing TM/MF management of cholesteatoma. RESULTS: Of 488 patients with cholesteatoma treated by the otological service between 1984 and 1998, 11 patients underwent TM/MF exposure and removal of anterior epitympanic cholesteatoma. Total cholesteatoma removal was accomplished in six patients. In three patients, because of facial nerve involvement, labyrinthine fistulae, or internal carotid artery involvement, open-cavity surgery was performed. In two patients, residual or recurrent cholesteatoma was exteriorized at "second-look" procedures. In this small cohort of patients the majority had extension to the arcuate eminence, geniculate ganglion, or Kawase's triangle or had "blue-lining" of the cochlea or labyrinth. To a lesser degree, the middle ear and mastoid contents were involved. Further facial nerve dysfunction or sensorineural hearing loss was not noted after surgery. CONCLUSIONS: Selective TM/MF removal of cholesteatoma provides an optimal route for removing complex cholesteatoma in patients with intact sensorineural function and medial cholesteatoma extension. PMID- 11037822 TI - A review of facial nerve outcome in 100 consecutive cases of acoustic tumor surgery. AB - OBJECTIVE: To determine the facial nerve outcomes at a tertiary neurotological referral center specializing in acoustic neuroma and skull base surgery. STUDY DESIGN: Retrospective review of 100 consecutive patients in whom acoustic neuromas were removed using all of the standard surgical approaches. METHODS: Functional facial nerve outcomes were independently assessed using the House Brackmann facial nerve grading system. RESULTS: The tumors were categorized as small, medium, large, and giant. If one excludes the three patients with preoperative facial palsies, 100% of the small tumors, 98.6% of the medium tumors, 100% of the large tumors, and 71% of the giant tumors had facial nerve function grade I-II/VI after surgery. CONCLUSION: Facial nerve results from alternative nonsurgical treatments must be compared with facial nerve outcomes from experienced surgical centers. Based on the facial nerve outcomes from our 100 consecutive patients, microsurgical resection remains the preferred treatment modality for acoustic tumors. PMID- 11037823 TI - Sensorineural and conductive hearing loss associated with lateral semicircular canal malformation. AB - OBJECTIVE: Lateral semicircular canal (LSCC) malformation is one of the most common radiological inner ear malformations. Traditionally, inner ear malformations are thought to be associated with sensorineural hearing loss (SNHL). Recent experience with patients with LSCC malformation suggested that LSCC malformation may be associated with both SNHL and conductive hearing loss (CHL). The auditory phenotype associated with LSCC malformation is not well delineated. The objective of this study is to define the nature of the hearing loss associated with LSCC malformation. STUDY DESIGN: Retrospective review METHODS: Retrospective review of clinical records, audiological evaluation, and imaging studies. RESULTS: Two patients with unilateral and 13 patients with bilateral LSCC malformation were identified. LSCC malformation was associated with CHL in 14% (4 ears), SNHL in 71% (20 ears), normal hearing in 11% (3 ears) and CHL due to atresia in one ear. Hearing loss varied from mild to profound but did not correlate with the severity of LSCC malformation. In bilateral malformation, the hearing loss was asymmetric in half of the cases. Malformation of the posterior limb of the LSCC was always associated with a large vestibular aqueduct. An absent or rudimentary LSCC was invariably associated with a cochlear abnormality. CONCLUSIONS: LSCC malformation, like other inner ear malformations such as large vestibular aqueduct and X-linked mixed deafness with perilymph gusher, can be associated with CHL, SNHL, or normal hearing. Children with unexplained conductive hearing loss often undergo exploratory surgery to improve hearing. Given that inner ear malformations may be associated with a pure CHL, it is critical that children undergo computed tomography scan of the temporal bone prior to undergoing exploratory surgery. PMID- 11037824 TI - Treatments for snoring: a comparison of somnoplasty and an oral appliance. AB - OBJECTIVES: Objectives of the study are 1) to test the effectiveness of somnoplasty (radiofrequency volumetric tissue reduction of the soft palate) for the control of loud, socially disruptive, snoring; 2) to test the long-term efficacy of this treatment by spouse report; and 3) to compare the effectiveness of somnoplasty with another treatment. STUDY DESIGN: Twenty patients with complaints of loud snoring and a respiratory disturbance index no greater than 15 respiratory events per hour with sleep-related episodes of oxygen desaturation no lower than 80% were offered an experimental treatment. Ten were treated with somnoplasty, and a comparison group of 10 matched patients used an oral appliance. All were restudied in the laboratory wearing a device programmed to count minutes of sleep during which snoring was loud, soft, or absent. METHODS: For the 10 somnoplasty patients, a spouse rating of snoring determined whether the patient received one or more treatments. Five patients had a single treatment to three sites, and five others had two such treatments. Ten patients wore an oral appliance of the tongue-retaining type (Snore X, Fremont, CA). RESULTS: Seven of the 10 somnoplasty patients met the improvement criteria set for reduced loud snoring (a spouse rating of 3 or less on a 10-point scale, and 10% or less of sleep time in loud snoring in the laboratory). The comparison group also had a significant improvement in the percentage of sleep time in loud snoring while wearing the Snore X appliance. CONCLUSION: Since there was no significant difference between the two groups in percentage of sleep time spent in loud snoring while treated, the choice of method to control snoring must be based on factors other than efficacy. PMID- 11037825 TI - Maxillomandibular advancement for persistent obstructive sleep apnea after phase I surgery in patients without maxillomandibular deficiency. AB - OBJECTIVE: To assess the outcomes of maxillomandibular advancement (MMA) for the treatment of persistent obstructive sleep apnea syndrome (OSA) after phase I reconstruction in patients who do not have maxillomandibular deficiency. METHODS: From January 1997 to September 1998, 25 patients previously treated with phase I reconstruction (uvulopalatoplasty, genioglossus advancement, and/or hyoid suspension) who did not have maxillary and mandibular deficiencies underwent MMA for persistent OSA. Variables examined include age, sex, body mass index (BMI), respiratory disturbance index (RDI), lowest oxygen saturation (LSAT), and cephalometric data. In addition, a minimum of 6 months after surgery, questionnaires containing a 10-cm visual analogue scale (0 = no change, 10 = drastic change) were mailed to the patients. The questionnaire subjectively assessed the patient's perception of the facial appearance after surgery, whether there was pain or discomfort of the temporomandibular joint, the overall satisfaction with the treatment outcomes, and whether the patient would recommend the operation to other patients. RESULTS: Nineteen (76%) questionnaires were completed and returned by 15 men and 4 women. The mean age was 45.3 +/- 6.6 years and the mean BMI was 33.1 +/- 7.1 kg/m2. The mean RDI improved from 63.6 +/- 20.8 to 8.1 +/- 5.9 events per hour, and the mean LSAT improved from 73.3 +/- 13.2% to 88.1 +/- 4.1%. One patient was defined as an incomplete responder (RDI >20). One patient reported transient pain and discomfort of the temporomandibular joint. Although all of the patients felt that there were changes in their facial appearance after surgery, 18 of the 19 patients gave either a neutral or a favorable response to their facial esthetic results. All of these patients were satisfied with the overall outcomes and would recommend the treatment to others. CONCLUSION: MMA is a highly effective treatment for persistent OSA after phase I surgery in patients who otherwise do not have maxillomandibular deficiency. The patient satisfaction is extremely high. Furthermore, previous concerns of unfavorable postoperative facial esthetics and temporomandibular joint dysfunction do not appear to be significant. PMID- 11037826 TI - Obstructive sleep apnea syndrome: a comparison between Far-East Asian and white men. AB - OBJECTIVES: To investigate the possible differences between Far-East Asian men and white men in obstructive sleep apnea syndrome (OSAS). STUDY DESIGN: Prospective nonrandomized controlled study. METHODS: This study compared consecutive Far-East Asian men with OSAS (n = 50) with two selected groups of White men with OSAS (n = 50 in each group). One group of white men was controlled for age, respiratory disturbance index (RDI), and minimum oxygenation saturation (LSAT). Another group was controlled for age and body mass index (BMI). Cephalometric analysis was performed on all subjects. RESULTS: The majority of the Far-East Asian men were found to be nonobese (mean BMI, 26.7 +/- 3.8) but had severe OSAS (mean RDI, 55.1 +/- 35.1). When controlled for age, RDI, and LSAT, the white men were substantially more obese (mean BMI, 29.7 +/- 5.8, P = .0055). When controlled for age and BMI, the white men had less severe illness (RDI, 34.1 +/- 17.9, P = .0001). Although the posterior airway space and the distance from the mandibular plane to hyoid bone were less abnormal in the Far-East Asian men, the cranial base dimensions were significantly decreased. CONCLUSIONS: The majority of the Far-East Asian men in this study were found to be nonobese, despite the presence of severe OSAS. When compared with white men, Far-East Asian men were less obese but had greater severity of OSAS. There may be differences in obesity and craniofacial anatomy as risk factors in these two groups. PMID- 11037827 TI - Engineering autogenous cartilage in the shape of a helix using an injectable hydrogel scaffold. AB - OBJECTIVE: Previous successful efforts to tissue engineer cartilage for an auricle have used an immunocompromised nude mouse xenograft model. Subsequent efforts in an immunocompetent autogenous animal model have been less successful because of an inflammatory response directed against the foreign scaffold polymer used to provide an auricular shape. We studied an alternative polymer material and surgical technique to engineer autogenous cartilage in the shape of a human ear helix using injectable hydrogel scaffolding, Pluronic F-127 (polyethylene oxide and polypropylene oxide). SUBJECT: Yorkshire swine. MATERIAL AND METHODS: Fresh autogenous chondrocytes were suspended in a biodegradable, biocompatible co polymer hydrogel, Pluronic F-127, at a concentration of 3 x 10(7) cells/mL. To support the contour of the implant, a skin fold channel in the shape of the helix of a human ear was created in the skin in three sites on the ventral surface of the animal. The cell-hydrogel suspension was injected through the skin fold channel. For controls, injections were made into identical channels using either cells alone or the Pluronic F-127 without cells. After 10 weeks, the specimens were excised and examined both grossly and histologically. RESULTS: Grossly, all implants retained a helical-like shape. Excised specimens possessed flexible characteristics consistent with elastic cartilage. The specimens could be folded and twisted and on release of mechanical pressure would instantly return to the original shape. Histological evaluation of the implants using H&E, Safranin O, trichrome blue, and Verhoeff's stains demonstrated findings consistent with mature elastic cartilage. Control injection of hydrogel alone demonstrated no evidence of cartilage formation and control injection of chondrocytes alone showed evidence only of disassociated elastic cartilage. CONCLUSION: Injection of autologous porcine auricular chondrocytes suspended in a biodegradable, biocompatible hydrogel of Pluronic F-127 resulted in the formation of cartilage tissue in the approximate size and shape of a human ear helix. This preliminary method extends the concept of auricular tissue engineering from an immunocompromised xenograft animal model to an immunocompetent autologous animal model. PMID- 11037828 TI - Otolaryngology consultation for peritonsillar abscess in the pediatric population. AB - OBJECTIVE: To assess clinical outcomes of children seen in consultation for peritonsillar abscess treated without the routine use of computed tomography or needle aspiration. STUDY DESIGN: Retrospective review of patients evaluated in the emergency department for possible peritonsillar abscess. Patient outcomes are reviewed with a statistical analysis of children grouped according to age. METHODS: A series of 102 patients, ages 8 months to 19 years, who were evaluated by the emergency department with otolaryngology consultation for possible peritonsillar abscess. All patients were admitted and given intravenous fluid replacement, antibiotics, and analgesia. Patients who responded to 24 hours of medical treatment were discharged, whereas patients who did not respond underwent elective tonsillectomy. MAIN OUTCOME MEASURE: Outcome of patients evaluated for peritonsillar abscess treated without immediate surgery, needle aspiration, or computed tomography. Outcomes are correlated with age and clinical findings. RESULTS: Fifty-two patients were discharged after initial medical therapy. Fifty patients underwent elective tonsillectomy; 40 of these patients were found to have abscesses at the time of surgery. When analyzed according to age, patients ages 8 months to 6 years were more likely to respond to medical treatment than children ages 7 to 12 and 12 to 19 (P = .023). Significant differences in the mean age of children requiring surgery (11.0 y) compared with those who responded to medical treatment (7.9 y) were observed (P = .003). Younger children who underwent tonsillectomy had a lower incidence of surgically confirmed abscess. CONCLUSIONS: A significant number of children presenting with odynophagia, malaise, pharyngotonsillar bulge, and decreased oral intake respond to medical therapy without radiological evaluation or surgical intervention. Additionally, younger children (1-6 y) are more likely to respond to medical treatment than older children. Pertinent clinical data, as well as advantages and disadvantages of this approach, are discussed. PMID- 11037829 TI - Value of antral puncture in the intensive care patient with fever of unknown origin. AB - OBJECTIVE: To evaluate the use of maxillary sinus puncture as a routine diagnostic procedure to exclude or confirm purulent sinusitis in intensive care unit (ICU) patients presenting with fever or a septic state of unknown origin. STUDY DESIGN: Retrospective. METHODS: All patients admitted to the ICU at the University Hospital Ghent who required ENT examination to exclude acute sinusitis as possible cause of their otherwise inexplicable fever or septic state underwent maxillary sinus puncture via the inferior meatus. The results of clinical examination and the relation between the presence of foreign bodies (e.g., nasogastric tubes) and culture results from the middle meatus and sinuses were analyzed. RESULTS: One hundred five punctures were performed in 53 patients. Macroscopic purulent effusions were obtained from 25 and nonpurulent effusions from 19 sinuses. The presence of a nasogastric tube did not influence puncture results but significantly increased colonization of the middle meatus. Staphylococcus aureus and Gram-negative agents were frequently cultured from sinus aspirates. Although purulent secretions often reveal no growth, most patients present with a multibacterial (40%) or monobacterial (28%) infection. Simple anterior rhinoscopy reduces the need for antral puncture. Only 8% of punctures in patients with a normal clinical examination were positive. CONCLUSIONS: Antral puncture proves to be a simple, fast, safe, inexpensive, and effective procedure for immediate diagnosis of acute nosocomial sinusitis in ICU patients and is therefore recommended as first procedure in these patients, even when only minor clinical abnormalities are present. PMID- 11037830 TI - Sinonasal disease and olfactory impairment in HIV disease: endoscopic sinus surgery and outcome measures. AB - OBJECTIVES: To investigate the effects of ESS (ESS) on olfactory impairment, disease-specific symptoms, and general quality of well-being In HIV+ patients with sinonasal disease. STUDY DESIGN: Study 1: Nasal cytology, rhinomanometry, nasal examination including endoscopy, disease-specific sinonasal symptoms, olfactory threshold sensitivity, and odor identification testing were performed before and after ESS in HIV+ patients with sinonasal complaints. Study 2: Quality of well-being was assessed before and after ESS in HIV+ patients with sinonasal complaints and controls. RESULTS: Significant olfactory sensitivity loss persisted for patients with chronic sinusitis after ESS, suggesting that the impairment in these patients may be due to viral disease rather than inflammation. Significant improvement in other disease-specific symptom scores (nasal obstruction, nasal congestion, headache, sinus pain, etc.) and results of the general quality of well-being assessment showed ESS to be beneficial in the extended health management of HIV illness. CONCLUSIONS: The current study indicated both olfactory dysfunction and subjective negative symptoms in HIV+ patients with chronic sinusitis. Although olfactory dysfunction remained, ESS was successful in providing marked alleviation of symptomatology in HIV+ patients with chronic sinusitis. The results support ESS as an appropriate treatment option for HIV+ patients with chronic sinusitis. PMID- 11037831 TI - Tumor necrosis factor gene polymorphism in chronic sinusitis. AB - OBJECTIVES: It is likely that genetic factors play a role in the etiology of chronic sinusitis, and airway inflammation is an important pathological feature in chronic sinusitis. We hypothesized that individuals with greater inflammatory responses may be more likely to acquire the disease. Polymorphisms of the tumor necrosis factor (TNF) genes have been described, and certain inflammatory diseases are reportedly associated with certain alleles of TNF genes. The purpose of this study is to examine whether there is an association between some alleles of TNF genes and chronic sinusitis. STUDY DESIGN: Thirty-eight Japanese patients with intractable chronic sinusitis were selected on the basis of the following criteria: 1) persistent mucous or mucopurulent nasal discharge and/or postnasal dripping for longer than 3 years and 2) opacification in bilateral maxillary sinuses and ethmoid cells on plain radiographic films. METHODS: Both tumor necrosis factor-alpha (TNF-alpha) and tumor necrosis factor-beta (TNF-beta) gene polymorphisms were analyzed by polymerase chain reaction (PCR) with restriction fragment length polymorphisms in these patients and 35 healthy control subjects. RESULTS: A significantly higher frequency (P < .05) of TNFB*2 allele of TNF-beta gene polymorphism was observed in patients with chronic sinusitis (74%) compared with control subjects (56%). There was no association between alleles of TNF alpha and chronic sinusitis. CONCLUSION: We concluded that TNF-beta gene polymorphism may form a component of the genetic predisposition to chronic sinusitis in Japanese patients. PMID- 11037832 TI - Clinical course of pediatric congenital inner ear malformations. AB - OBJECTIVE: To determine any factors that could improve the early detection and management of congenital inner ear malformations. STUDY DESIGN: A retrospective review was performed of all patients with a diagnosis of inner ear malformation at Loyola University Medical Center (LUMC) and the Hospital for Sick Children (HSC) between 1987 and 1995. Clinical records and audiometric data were accumulated. One neuroradiologist reviewed every temporal bone computed tomography (CT) scan. METHODS: Forty-six pediatric patients with congenital inner ear anomalies evaluated at two tertiary care hospitals. RESULTS: The average patient age at initial assessment was 25.7 months. A family history of hearing loss was noted in only five patients (12.8%). A major nonotological deformity was seen in 41% of patients. The average hearing threshold was 88 dB. All three patients with sudden hearing loss had vestibular aqueduct enlargement. Two of the three patients with common cavity anomalies had a history of recurrent meningitis. Twenty-seven patients had a vestibular aqueduct deformity, the most frequent radiographic abnormality in the series. CONCLUSIONS: Because inner ear malformation was diagnosed after 24 months of age in a significant percentage of patients, we recommend increased parental education and vigilance by primary care practitioners. Universal newborn screening may be the key to earlier detection of these infants. For children with idiopathic sensorineural hearing loss, we recommend a temporal bone CT scan. Patients with vestibular aqueduct enlargement must be counseled about the risk of progressive sensorineural hearing loss, meningitis, and the need to avoid contact sports. Patients with common cavity abnormalities should be considered for exploratory tympanotomy and also educated about the risk for meningitis. PMID- 11037833 TI - Vestibular schwannoma growth: the continuing controversy. AB - OBJECTIVE: To investigate the growth of vestibular schwannoma (VS) in a series of 123 patients with 127 tumors allocated to the "wait and scan" group in the period 1973-1999. STUDY DESIGN: Retrospective review of prospectively registered data on all patients with VSIE from the entire country who were allocated to the wait and scan group. METHOD: Clinical charts, audiometric data, and neuroradiological images were reviewed and tabulated for age, hearing level expressed as speech reception threshold (SRT) and speech discrimination score (SDS), maximum extra canalicular tumor extension, and possible changes in tumor diameter. The material was updated three times (in June 1993, June 1996, and June 1999). Via the Danish national register, data on whether the included patients were alive or dead were collected in 1999. RESULTS: The tumor growth, growth rate, and growth patterns were calculated in three periods 1973 to 1993 (mean observation period, 3.4 y), 1973 to 1996 (mean observation period, 3.8 y), and from 1973 to 1999 (mean observation period, 4.2 y). By termination of the first period, 94 tumors (74%) exhibited measurable growth, 23 tumors (18%) no measurable growth, and 10 tumors (8%) revealed negative growth. By the end of the extended observation period, tumor growth was observed in 104 tumors (82%), no tumor growth in 15 tumors (12%), and negative growth in 8 tumors (6%). Subsequent to the third observation period, growth was observed in 108 tumors (85%), no growth in 11 tumors (9%) and negative growth in 8 tumors (6%). However, the results may also be interpreted in another way: 52 patients (42%) were alive at the time of writing, tumor growth did not demand any intervention, 23 patients (19%) died as a result of non-tumor related causes, and 35 patients (28%) were previously treated and alive by the termination of the third observation period. CONCLUSION: Depending on the observation period, three sets of growth results were obtained. The long observation period, updating and re-updating the results, gave us the opportunity for a de novo interpretation of the results and the long-term consequences of the wait and scan policy. Combined with other factors, the achieved results should be considered when timing of surgery is to be decided. PMID- 11037834 TI - Quantitative analysis of tympanic membrane disease using video-otoscopy. AB - OBJECTIVE: To perform quantitative analysis of pathological changes in the tympanic membrane using video-otoscopic images. STUDY DESIGN: Prospective case control study. METHODS: Forty-two ears of children with chronic otitis media with effusion (OME) and 15 ears of normal children were included in this study. Tympanic membrane images were captured and digitized using a Welch-Allyn (Skaneatales Falls, NY) VDX-300 Illumination and Imaging system with S-VHS input to a MIRO DC 30 (Pinnacle Systems, Mountain View, CA) visual board in a Power PC based computer. These images were visualized and recorded during static and pneumatic pressure changes. Quantitative analysis of tympanic membrane disease was performed using Image Pro Plus Imaging software (Media Cybernetics, Del Mar, CA). The measurements included area of the tympanic membrane and its quadrants, area of tympanic membrane involved by disease, angle formed at the umbo, and length of the malleus versus vertical length of the tympanic membrane. RESULTS: Tympanosclerosis was present in 57% of ears and occurred most frequently in the anteroinferior quadrant, but the maximum area of involvement was in the posteroinferior quadrant. The ratio of the angles formed at the umbo was significantly greater (P = .01) for the OME group compared with the control group. The ratio of the length of the umbo and the vertical length of the tympanic membrane was almost identical for the OME and the control groups (P = .4). CONCLUSIONS: Videootoscopic images can be used for quantitative analysis of tympanic membrane disease. The ratio of the posterior angle to the anterior angle formed at the umbo seems to be a more reliable indicator of post otitis media than is a reduced length of the long process of malleus. PMID- 11037835 TI - A clinicopathological study of 15 patients with neuroglial heterotopias and encephaloceles of the middle ear and mastoid region. AB - OBJECTIVES/HYPOTHESIS: Heterotopic masses of neuroglial tissue involving non midline structures, specifically, the middle ear region, are exceptional. The pathogenesis of these lesions and, in particular, their relation to encephaloceles, is uncertain. STUDY DESIGN AND METHODS: H&E-stained sections from 15 lesions diagnosed as neuroglial heterotopias or encephaloceles involving the middle ear region were reviewed. Radiographic or operative evidence of a central nervous system (CNS) relation and clinical factors possibly related to pathogenesis were analyzed. RESULTS: All 15 lesions (from six men and nine women; mean age, 49 y; range, 16-67 y), regardless of their relation to the CNS, were composed of varying proportions of neurons and glia with associated chronic inflammatory cells and reactive gliosis. No significant ependymal or choroid plexus component was present. Operative findings revealed that two lesions had definite CNS connections and two were unrelated to the CNS; this relation could not be determined in the remaining cases. Seven of 10 patients for whom clinical information was available had a history of chronic otitis media or mastoiditis or both; four of these seven patients also had a history of previous trauma or surgery. Three patients, including both patients whose lesions had no demonstrable CNS attachment, had no predisposing factors. CONCLUSIONS: Most neuroglial heterotopias of the middle ear are probably acquired encephaloceles. These lesions occur in older patients than do their midline counterparts. Determination of the relation of these lesions to adjacent CNS structures must be done radiographically or using operative findings, because histology alone cannot be reliably used to render an accurate diagnosis. PMID- 11037836 TI - Influence of serum lipids on auditory function. AB - OBJECTIVE: To evaluate the influence of serum lipids on auditory function. STUDY DESIGN: The study group comprised 607 men and 317 women who underwent medical examination at our hospital. Eligibility criteria included 1) age of 40 to 59 years; 2) normal ear drums; 3) no history of noise exposure or of diseases associated with hearing loss; 4) normal results of a glucose tolerance test; and 5) normal hearing or sensorineural hearing loss with a flat-form or high-tone gradual-form audiogram. We investigated the relation between the hearing level of the better-hearing ear and serum concentrations of total cholesterol, triglyceride, and high-density lipoprotein cholesterol. METHODS: For each variable, the subjects were divided into two groups--a high-level group (serum lipid concentration > or = 1 SD higher than the mean) and a low-level group (serum lipid concentration < or = 1 SD lower than the mean). Differences in hearing level between the two groups were compared according to sex with the t test. RESULTS: For total cholesterol and total triglyceride, there was no significant difference in hearing level between the high-level group and the low level group in either sex. As for high-density lipoprotein cholesterol, hearing levels at 2,000 Hz (P < .05) and 4,000 Hz (P < .01) in the high-level group were significantly better than those in the low-level group in men. CONCLUSION: A low high-density lipoprotein cholesterol concentration is associated with hearing loss. Arteriopathy may play a role in auditory dysfunction. PMID- 11037837 TI - Influenza A virus infection of human middle ear cells in vitro. AB - HYPOTHESIS: Human-derived normal middle ear mucosal cells can be harvested and cultured and will support influenza A virus (INF A) infection. STUDY DESIGN: Protocols for the collection and in vitro culture of middle ear mucosal cells were developed and used to investigate the effects of INF A infection as it relates to the pathogenesis of otitis media. MATERIALS AND METHODS: Middle ear mucosa was harvested during surgeries that opened the normal middle ear. Middle ear mucosal cells were plated and grown in collagen-coated dishes. Cells were characterized before and after INF A exposure using phase-contrast and immunofluorescence microscopy as well as reverse transcriptase-polymerase chain reaction (RT-PCR) for cytokeratin 18 gene expression and INF A. RESULTS: Primary cultures of human middle ear epithelial cells were established. Prolonged growth of middle ear cells yielded a second cell type that failed to stain for cytokeratin on immunofluorescence but continued to produce positive RT-PCR results on cytokeratin 18 analysis. After INF A exposure, cytological changes and immunofluorescence staining showed cellular infection. RT-PCR analysis using INF A-specific primers showed positive results for up to 72 hours after viral exposure. CONCLUSIONS: Primary cultures of human middle ear mucosal cells have been established. Two distinctly different cell culture systems have been developed: 1) middle ear epithelial cells and 2) either dedifferentiated epithelial cells or fibroblasts. Exposure of both cell types to INF A demonstrates that each can support cellular infection and viral replication. These models should be useful for studies of the pathogenesis of virus-mediated otitis media. PMID- 11037839 TI - New options for aerodigestive tract replacement after extended pharyngolaryngectomy. AB - OBJECTIVES: Assess the results of a new type of reconstruction of the aerodigestive tract after extended pharyngolaryngectomy. STUDY DESIGN: Follow-up of a total of eight patients who had surgery using ileocolic free graft. METHODS: The surgical technique is described. Five patients underwent pharyngolaryngectomy/cervical esophagectomy, and three patients had total laryngectomy with subtotal pharyngectomy. Patients were monitored to assess complications and recovery of satisfactory swallowing and speech. RESULTS: The technique, thanks to the use of material from the colon, proved to be extremely useful for the reconstruction of the digestive tract. At the same time, ileal anastomosis with the tracheal stump enabled aerodigestive crossing restoration, protected by the ileocecal valve. All patients recovered good swallowing capacity and phoniatric expression, which were obtained by digital occlusion of the tracheostomy, forcing the expiratory air through the ileum and ileocecal valve. Manometric tests also showed that after a while there was a gradual synchronization of swallowing between the transplanted colic segment and the residual esophagus. CONCLUSIONS: The technique described in the present study may be regarded, also in the light of possible further applications, as a new and interesting option for the reconstruction of the aerodigestive tract. PMID- 11037838 TI - Complement C3 cleavage and cytokines interleukin-1beta and tumor necrosis factor alpha in otitis media with effusion. AB - OBJECTIVES: To analyze whether complement C3a anaphylatoxin, other C3 fragments, interleukin-1beta (IL-1beta), or tumor necrosis factor-alpha (TNF-alpha) contributes to inflammation in chronic otitis media with effusion (OME). METHODS: The amount of C3a was measured by enzyme-linked immunoassay. Further breakdown of C3 was analyzed by Western blotting. IL-1beta and TNF-alpha concentrations were measured by radioimmunoassay. Bacteria were analyzed by culture and polymerase chain reaction. RESULTS: Highly elevated levels of C3a and other C3 cleavage fragments were found in all middle ear effusion (MEE) samples. The mean values (+/- SEM, n = 26) for C3a, IL-1beta, and TNF-alpha were 5,973 +/- 1,124 ng/mL, 1,043 +/- 490 pg/mL, and 79 +/- 14.3 pg/mL, respectively. Comparison to an average C3 level of 555 (+/-108) microg/mL indicated that at least 40.5% +/- 6% of total C3 had become activated within the MEE. C3a concentrations were higher in the group in which the effusion had been present in the middle ear for a prolonged period (> or =4 mo) (P = .04). Children with multiple tube insertions had higher C3 (P = .006) and TNF-alpha (P = .04) concentrations in their MEE samples than those receiving their first tubes. C3 and C3a concentrations in MEE correlated to each other (correlation coefficient [r] = 0.513, P = .0056), as did concentrations of IL-1beta and TNF-alpha (r = 0.7016, P < .0001). No significant correlation was found between complement C3 or C3a levels and IL-1beta, TNF alpha, or bacterial growth. CONCLUSIONS: Highly elevated levels of C3a in MEE indicate ongoing complement activation, which is stronger than in almost any other disease demonstrated previously. Elevated C3a levels contribute to chemotactic and inflammatory potential in the MEE and correlate with the chronicity of the disease. PMID- 11037840 TI - Raman spectroscopy for early detection of laryngeal malignancy: preliminary results. AB - OBJECTIVE: Raman spectroscopy, the analysis of scattered photons after monochromatic laser excitation, is well established in nonbiological sciences. Recently this method has been used to differentiate premalignant and malignant lesions from normal tissue. Its application for early diagnosis has been explored in a variety of sites (e.g., esophagus, cervix), but not, to date, in laryngeal cancer. The objective of this study was to perform a feasibility study of the use of Raman spectroscopy for early diagnosis of laryngeal malignancy. METHODS: Biopsy specimens were snap-frozen, and top sections were sent for histopathological analysis. Only homogenous samples with clearly defined pathological findings were used in this study: seven histologically normal samples, four exhibiting dysplasia, and four with carcinoma. Samples were defrosted and placed under a Renishaw (Wotton-Under-Edge, UK) System 1000 Raman microspectrometer for analysis. Between 5 and 12 spectra were acquired from each sample, with an excitation wavelength of 830 nm. Average characteristic spectra for each disease or condition were compared. Further multivariate statistical analysis of the data was carried out to evaluate and maximize the differences in the spectra for each disease or condition. RESULTS: The most visible differences in the spectra occur between 850 and 950 cm(-1) and 1,200 and 1,350 cm(-1). The later peaks are directly related to protein conformation and C-H bond stretch in nucleic acid bases. The relative intensity of the nucleic acid peak increases with progression to malignancy. Use of linear discriminant analysis made it possible to separate the spectra with disease to a greater degree of accuracy than using empirical peak ratio methods alone. Classification results obtained from cross-validation of the discriminant model showed prediction sensitivities of 83%, 76%, and 92% and specificities of 94%, 91%, and 90% for normal, dysplastic, and squamous cell carcinoma of the larynx, respectively. CONCLUSIONS: There was strong evidence to support spectral identification of malignancy and earlier abnormal changes. More substantive studies of the spectral differences between malignant and non-neoplastic tissue are warranted. Raman spectroscopy may become a useful adjunct to pathological diagnosis allowing directed or guided biopsies and assessment of adequacy of resection margins. PMID- 11037841 TI - Analysis of treatment results for floor-of-mouth cancer. AB - OBJECTIVE: This study reports the results of treating floor-of-mouth cancer with five different treatment modalities with long-term follow-up. STUDY DESIGN: Retrospective study of 280 patients with floor-of-mouth cancer treated in the Department of Otolaryngology-Head and Neck Surgery at Washington University Medical School (St. Louis, MO) from 1960 to 1994. METHODS: Patients with biopsy proven squamous cell carcinoma of the floor of mouth who were previously untreated were treated with curative intent by one of five modalities and were all eligible for 5-year follow-up. The treatment modalities included local resection alone, composite resection alone (with neck dissection), radiation therapy alone, local resection with radiation therapy, and composite resection with radiation therapy. Multiple diagnostic, treatment, and follow-up parameters were studied using standard statistical analysis to determine statistical significance. RESULTS: The overall 5-year disease-specific survival (DSS) was 56% with death due to tumor in 44% of patients. The 5-year cumulative disease specific survival (CDSS) was 0.61 (Kaplan-Meier probability) with a mean of 8.3 years and a median of 9.7 years. The DSS by treatment modality included local resection (76%), composite resection (63%), radiation therapy (43%), local resection with radiation therapy (61%), and composite resection with radiation therapy (55%). Overall, there was no significant difference in DSS by treatment modality. Recurrence at the primary site (41%) was the most common site of treatment failure. Nineteen percent of patients had recurrence in the neck. Eighty-eight percent of initial recurrences occurred within 60 months after the onset of treatment. Metastasis to a distant site occurred in 30% of patients. Twenty percent of these patients had second primary cancers, and 53% of these patients died of their second primary cancers. CONCLUSIONS: Significantly improved 5-year DSS was seen in the patients with clear margins, early clinical tumor stage, and negative nodes. Significantly decreased 5-year survival was seen in the patients with involved margins, advanced clinical tumor stage, positive nodes, and tumor recurrence. Patients with no clinically positive nodes (cNO) can be observed safely for regional nodal disease and subsequent positive nodes can be treated as they occur with no adverse affect on survival. Because of high recurrence rates at the primary site and neck, and an increased rate of both distant metastasis and the development of second primary cancers, patients should be monitored closely for a minimum of at least 5 years. PMID- 11037842 TI - Accuracy, utility, and cost of frozen section margins in head and neck cancer surgery. AB - OBJECTIVES: Intraoperative frozen section analysis of surgical margins is widely used in head and neck cancer surgery. This study evaluates frozen section accuracy relative to permanent controls and final margins from the entire specimen, the rate at which frozen sections impact intraoperative management, and the resultant cost. STUDY DESIGN: Retrospective. METHODS: From 1997 to 1999 the frozen section results, permanent controls, and final tumor margins from 80 consecutive patients undergoing 420 intraoperative frozen section margins for head and neck malignancy were reviewed. RESULTS: A 98.3% accuracy rate (sensitivity, 88.8%; specificity, 98.9%) was found compared with permanent sections of the same tissue. However, 40% (8 of 20) of patients with positive final margins on the resection specimen, and 100% (15 of 15) with close (<5 mm) margins were not detected by frozen section analysis. The overall accuracy of frozen section in the evaluation of close or positive final margins was 71.3% (sensitivity, 34.3%; specificity, 100%). In addition, 5% (4 of 80) of patients potentially benefited from intraoperative frozen section by virtue of immediate margin revision. The estimated cost of intraoperative frozen section averaged as much as $3,123 per patient, with a cost-benefit ratio of 20:1. CONCLUSIONS: Intraoperative frozen section margins are accurate, but they are costly and cannot reliably eradicate positive final margins. Patients with early-stage lesions and those undergoing re-resection for recurrence or salvage surgery after radiation failure derived the greatest potential benefit from frozen section margins. PMID- 11037843 TI - Anesthesia for laryngeal surgery in the office. PMID- 11037844 TI - Intralaryngeal approach to laryngeal web using lateralization with silastic. PMID- 11037845 TI - A pilot series demonstrating fluorescence staining of laryngeal papilloma using 5 aminolevulinic acid. PMID- 11037846 TI - Fever and neutropenia: changing patterns of medical practice. PMID- 11037847 TI - Unconventional therapies: "eyes wide shut". PMID- 11037848 TI - Is adjuvant therapy ever warranted in localized neuroblastoma. PMID- 11037849 TI - Prognostic factors in acute lymphoblastic leukemia. PMID- 11037850 TI - Randomized placebo-controlled trial of oral antibiotics in pediatric oncology patients at low-risk with fever and neutropenia. AB - PURPOSE: Fever combined with neutropenia in pediatric oncology patients has traditionally been managed in the hospital with broad-spectrum intravenous antibiotics until there is documented neutrophil recovery. Recent evidence has suggested that patients at "low-risk" can be discharged from the hospital before neutrophil recovery. Whether oral antibiotics are required at the time of discharge is not known. PATIENTS AND METHODS: Using a randomized, double-blind, placebo-controlled study design, 73 patients at low-risk with episodes of fever and neutropenia were discharged home while still neutropenic: 37 administered with oral cloxacillin and cefixime and 36 administered with corresponding placebos. Low-risk criteria included: afebrile for more than 24 hours, negative blood culture results at 48 hours, absence of clinical sepsis, cancer in bone marrow remission, and absence of comorbid conditions. RESULTS: Five patients (14%; 95% confidence interval [CI]; 2%-25%) in the antibiotic arm and two patients (6%; 95% CI; 0%-13%) in the placebo arm were readmitted to the hospital with recurrent fever while still neutropenic (P = 0.43). One patient randomized to the placebo arm had a positive blood culture result on readmission, which responded to appropriate intravenous antibiotics. All of the readmissions were uneventful and there were no fatalities. The average cost per episode of fever and neutropenia was $1,821 Canadian dollars with only minimal incremental cost to the antibiotic arm. CONCLUSION: This study supports the discontinuation of antibiotics in pediatric oncology patients at low-risk who still have neutropenia at the time of discharge from the hospital. PMID- 11037851 TI - Use of unconventional therapies by children with cancer at an urban medical center. AB - PURPOSE: The aim of this study was to determine the prevalence, modalities, and determinants of use of unconventional therapies among children with cancer receiving conventional treatment at an urban academic medical center in the United States. PATIENTS AND METHODS: We interviewed the parents of patients and/or patients who were receiving or had received conventional therapy for treatment of childhood cancer. Of 78 patients/ parents asked, 75 consented to the interview, which included demographic factors, life events, and use of unconventional therapies. All participants also consented to the abstraction of chart data for this study. RESULTS: Overall, 84% of respondents reported the use of one or more unconventional therapies. The most commonly used modalities were changes in diet, nutritional and herbal agents, and mind/body treatments. Most users had tried more than one unconventional modality. No difference in use was seen by cancer diagnosis, race/ethnicity, socio-economic status, or educational attainment of the respondent. Of the therapies used, 50% were not reported to the physicians. Of patients reporting use of an unconventional approach, 85% were concurrently enrolled on clinical trials for primary treatment of their cancer. CONCLUSIONS: The use of unconventional therapies is highly prevalent among children with cancer and is not associated with demographic or clinical factors or participation in clinical trials. The possibility that an unconventional treatment may interact with a protocol treatment underscores the need for more information about the use of such therapies among all patients. PMID- 11037852 TI - Self-image of adolescent survivors of long-term childhood leukemia. AB - The purpose of our research was to evaluate the attitude to face the life cycle and the impact that the experience of childhood leukemia may have had in a group of adolescents who had the disease cured. A questionnaire was administered at the Pediatric Hematology Center, San Gerardo Hospital, Monza, Italy, to all former patients age 12 to 20 years and off therapy from leukemia for at least 2 years (total of 116 adolescents) during 1997; 70 patients responded to the mailing and a comparison group of 70 secondary-school students was investigated. The two groups were matched as closely as possible on key characteristics (age, gender, socio-economic level of families, education and occupation of the parents, and geographic area of residence). The Offer Self-Image Questionnaire was the instrument used in this study. Overall, the teenagers in whom leukemia was cured showed a more positive and mature self-image (psychologic, social, attitude toward family, and coping) compared with the student group (statistical evidence, P < 0.05). An effective psychosocial support for patients and their families during their treatment, in addition to medical therapy, is strongly recommended. The majority of survivors of childhood cancer grow successfully without serious psychologic sequelae. PMID- 11037853 TI - Malignant ovarian tumors in children: 22 years of experience at a single institution. AB - PURPOSE: Malignant ovarian tumors of childhood are relatively rare and thus, management is still unclear. We reviewed our experience with these tumors to evaluate their histopathologic characteristics, treatment, and outcome. PATIENTS AND METHODS: From January 1975 to December 1997, 56 patients had their malignant ovarian tumors diagnosed, treated, and followed-up in our institution. All tumors were completely excised when possible; otherwise, biopsy was performed. Staging was made according to Federation Internationale de Gynecologie Oncologique classification. Chemotherapy was recommended for all patients. Twelve cases were treated with vincristine, actinomycin, cyclophosphamide (VAC) before 1986; 12 with cisplatin, vinblastine, and bleomycin (PVB) from 1986 to 1989; and 23 with the bleomycin, etoposide, and cisplatin (BEP) regimen from 1989 to present. The Kaplan-Meier survival method was used to calculate the survival. The log-rank test was used to compare groups with respect to survival. RESULTS: Age range was 0 to 16 years (median 11 yrs; average 9.8 yrs). Only two patients were younger than 1 year. The most common presenting symptom was abdominal pain, occurring in 27 patients (48.2%). Thirty-three patients (60%) had total one-sided salpingo oophorectomy and three patients had bilateral salpingo-oophorectomy. Nineteen patients had stage I, 15 had stage II, 19 had stage III, and 3 had stage IV disease. Dysgerminoma was the most common type. Overall survival (OAS) and event free survival were 68% (median follow-up time: 71 mos) and 57%, respectively, after 22 years. Histopathology was not correlated with survival. Two important predictors for survival are age (P < 0.0001) and treatment protocol (P = 0.013). The BEP protocol was superior to the other regimens. The OAS was 74.6% in BEP, 55% in PVB, and 63.6% in VAC regimens. CONCLUSION: Although age at diagnosis and treatment with BEP regimen have major roles in determining prognosis of the ovarian tumors in childhood, for patients with advanced ovarian germ cell tumors, intensification of chemotherapy or the development of new approaches is necessary. PMID- 11037854 TI - Pediatric melanoma: are recent advances in the management of adult melanoma relevant to the pediatric population. AB - PURPOSE: Although melanoma in childhood is a rare condition, there is evidence that it is increasing in frequency. As advances are being made in the understanding and therapy of adult melanoma, we need to consider the relevance of these advances to the pediatric population. PATIENTS AND METHODS: We have reviewed our experience at the University of Colorado Health Sciences Center with the clinical parameters, therapy, and outcomes of melanoma in 27 patients age 16 years or younger and contrasted these to the adult experience. RESULTS: Most cases were diagnosed early with the median thickness of the primary melanoma being 0.75 mm. Six of seven patients who had lymph node metastases develop remain alive at a median follow-up of 62 months. Durable complete responses to a variety of therapies were seen in three of five patients with advanced disease outside the central nervous system. Our experience with sentinel node biopsy, adjuvant interferon, and new therapies for metastatic melanoma were also reviewed and appear to be relevant for younger patients. CONCLUSIONS: The behavior of melanoma in the pediatric population at our center is similar to that seen in adults. The integration of recent advances in the staging and therapy of melanoma in adults would be of benefit to children with this condition. PMID- 11037856 TI - Successful cord blood transplantation for sickle cell anemia from a sibling who is human leukocyte antigen-identical: implications for comprehensive care. AB - We report the successful transplantation of umbilical cord blood stem cells from a sibling who is human leukocyte antigen-matched to a child with sickle cell anemia. Conditioning was with busulfan, cyclophosphamide, and antithymocyte globulin. Time to neutrophil count >500/microL was 23 days and to platelet count >50,000/microL was 49 days. Full donor engraftment was achieved without graft versus-host disease. This case demonstrates the potential usefulness of harvesting cord blood from full siblings of patients with sickle cell disease. Routine collection of umbilical cord blood from siblings should be considered for patients with sickle cell disease, and may increase acceptance and use of transplantation by families. PMID- 11037855 TI - Fludarabine-based protocol for haploidentical peripheral blood stem cell transplantation in Hurler syndrome. AB - To assess the feasibility of performing a haploidentical peripheral blood stem cell transplantation (PBSCT) in a child with Hurler syndrome after a novel conditioning regimen consisting of fludarabine monophosphate, anti-T-lymphocyte globulin, low-dose busulfan, and single-dose total body irradiation of 750 cGy. A 16-month old boy with Hurler syndrome underwent haploidentical PBSCT from his 3/6 HLA-matched sister. Pretransplant conditioning consisted of fludarabine (30 mg/m2 per day) from day -10 to day -5, busulfan (4 mg/kg per day) on days -7 and -6, rabbit anti-T-lymphocyte globulin (10 mg/kg per day) from day -4 to day -1, and total body irradiation of 750 cGy on day -1. In vitro T-cell depletion was carried out with rat antihuman CDw52 monoclonal antibody (Campath-1G). The fludarabine-based protocol was well-tolerated, with mild toxicity and no major transplant-related complications or graft-versus-host disease. Engraftment was complete and stable. Chimerism was 100% donor origin, as determined by restriction fragment length polymorphism. Cytogenetic and polymerase chain reaction-various number of tandem repeats (PCR-VNTR) analyses of peripheral blood and bone marrow showed 100% reconstitution with female donor cells. The patient underwent the transplant 30 months ago and is in good clinical condition, with normal counts, no signs of graft-versus-host disease, and no infectious episodes; neurologic signs have stabilized. Haploidentical PBSCT, T-cell-depleted by means of Campath-1G, may serve as a therapeutic alternative for patients with Hurler syndrome when a fully matched sibling is not available. PMID- 11037857 TI - Severe 6-thioguanine-induced marrow aplasia in a child with acute lymphoblastic leukemia and inherited thiopurine methyltransferase deficiency. AB - 6-thioguanine (6TG) is undergoing investigation for use in the maintenance phase of acute lymphoblastic leukemia (ALL). Just as with 6-mercaptopurine (6MP), it is be expected that 6TG would cause pancytopenia in individuals with inherited thiopurine methyltransferase (TPMT) deficiency. We report the first case of severe and prolonged pancytopenia caused by 6-thioguanine in an 8-year-old boy with ALL and inherited TPMT deficiency. Neutropenia lasted 67 days, whereas anemia and thrombocytopenia did not recover for 96 days. To obviate this life threatening complication, clinicians should consider assaying TPMT activity before initiating therapy with 6MP and, particularly, 6TG in children with ALL. PMID- 11037858 TI - Successful dose-intensive treatment of desmoplastic small round cell tumor in three children. AB - Desmoplastic small round cell tumor (DSRCT) is a rare soft tissue tumor of primitive origin occurring primarily in children and young adults. Based on published reports in the literature, the response to conventional chemotherapy is poor. We report three pediatric patients successfully treated with dose intensive, multimodal therapy. Between August 1994 and March 1998, we evaluated three consecutive patients with DSRCT at Children's Hospital and Regional Medical Center, Seattle, Washington. We established the diagnosis based on clinical presentation, radiologic staging, and pathologic review with immunohistochemical staining. All patients received a combined modality protocol including dose intensive chemotherapy (two of them with peripheral blood stem cell [PBSC] support), second look surgery, and consolidative local irradiation. The patients remain in continuous remission at 66, 42, and 26 months after diagnosis, respectively. Two of our patients were younger than any previously reported patient, extending the age group for which DSRCT should be considered on diagnosis of small round cell tumors. The uniform survival achieved in our series indicates potential benefit for the combination of dose-intensive multiagent chemotherapy, local irradiation, and aggressive surgical approach in this disease. PMID- 11037859 TI - Concurrent acute lymphoblastic leukemia and juvenile pilocytic astrocytoma in a pediatric patient. AB - The concurrence of acute lymphoblastic leukemia (ALL) and an asymptomatic juvenile pilocytic astrocytoma is described. A 6-year-old boy without clinical evidence of neurofibromatosis had a juvenile pilocytic astrocytoma diagnosed on radiologic examination and before treatment of acute pre-B cell lymphoblastic leukemia. The patient has had a partial resection of the astrocytoma and is 9 months into treatment of his ALL, which is in complete remission. p53 gene mutation was not identified in this patient. The concurrent diagnosis before treatment of ALL and juvenile pilocytic astrocytoma, the latter normally an indolent tumor, suggests that some cases of astrocytoma previously ascribed to radiotherapy or other treatment may in fact be caused by other factors. PMID- 11037860 TI - Renal enlargement as presentation of isolated renal relapse in childhood leukemia. AB - Extramedullary relapses in children with acute lymphoblastic leukemia occur most frequently in the central nervous system and in the testis. In this report, the authors describe a 16-year-old girl with an isolated renal relapse of acute lymphoblastic leukemia after a disease-free interval of 2 years and 8 months. This clinically inconspicuous renal relapse was suggested by a routine follow-up renal sonography. No evidence of disease was found in bone marrow or peripheral blood. Renal biopsy was required to establish the diagnosis. Treatment consisted of intensive chemotherapy and autologous bone marrow transplantation. The patient has been in second complete continuous remission for 7 years. The authors recommend the use of an intensive multidrug salvage regimen. PMID- 11037861 TI - Diabetes insipidus as a presenting symptom of acute myelogenous leukemia. AB - This report describes a case of diabetes insipidus associated with acute myelogenous leukemia. An 11-year-old boy presented with fatigue, polydipsia and polyuria. His evaluation revealed a diagnosis of acute myelogenous leukemia FAB M2, and a water deprivation test confirmed the diagnosis of central diabetes insipidus. His brain magnetic resonance imaging (MRI) showed a thickened, enhancing pituitary stalk with absence of the normal hyperintense signal in the posterior pituitary. He was treated with systemic chemotherapy, intensive intrathecal therapy, and 1,000 cGy to the pituitary. The patient achieved a remission but continued to need desmopressin therapy to control his diabetes insipidus. Diabetes insipidus is a rare complication of acute myelogenous leukemia that can be caused by leukemic infiltration of the pituitary. The diabetes insipidus is irreversible despite intensive systemic and central nervous system chemotherapy and radiation. PMID- 11037862 TI - Fatal lymphoproliferative disease as a complication of Evans syndrome. AB - A 9-month-old boy had bruising and petechiae. Investigation revealed a Coombs positive hemolytic anemia and immune-mediated thrombocytopenia. The infant was treated with intravenous immunoglobulin and steroids. The infant eventually had recurrent fevers, hepatosplenomegaly, pulmonary nodules, and parenchymal central nervous system (CNS) lesions develop. Results of a lung biopsy revealed a polyclonal lymphoproliferative disease. Polymerase chain reaction analysis showed the presence of the Epstein-Barr (EB) viral genome in the lung nodules. The infant died from progressive lung disease 6 months after the initial symptoms of Evans syndrome. Lymphoproliferative disease is known to occur in a variety of settings after immunosuppression, especially in solid organ transplant recipients. We report a case of polyclonal lymphocyte proliferation in a patient with Evans syndrome. PMID- 11037863 TI - Lymphoid malignancy as a presenting sign of ataxia-telangiectasia. AB - Ataxia-telangiectasia (AT) is an uncommon genetic disorder characterized by cerebellar ataxia, oculocutaneous telangiectasias, progressive immunodeficiency, and a predisposition to lymphoid malignancy. The genetic defect in AT predisposes not only to malignancy but also to severe toxicity from anti-neoplastic therapies. It is important to consider the diagnosis of AT in any child with a lymphoid malignancy at a younger than expected age, or who has a pre-existing ataxia, to anticipate unusually severe toxicities from the antineoplastic therapy, to avoid confusing the development of ataxia with toxicity from therapy, and to provide appropriate genetic counseling. We describe two children at a young age with a lymphoid malignancy diagnosed before the diagnosis of AT. One patient had severe toxicity from his chemotherapy, requiring truncation of the planned course of treatment. The other child was able to tolerate his entire planned course of therapy, but ataxia that was initially interpreted as toxicity from chemotherapy rather than as a sign of his AT developed. Lymphoid malignancy may be the presenting sign of AT. Making this diagnosis may influence therapy of the malignancy. The neurologic manifestations of the disease can be misinterpreted as toxicities of the chemotherapy, and diagnosis of AT allows appropriate genetic counseling for the family. PMID- 11037864 TI - Lymph node manifestations of limited Churg-Strauss syndrome. AB - Churg-Strauss syndrome is a systemic vasculitis characterized by asthma, tissue and blood eosinophilia, and granulomatous vasculitis. Lymph node involvement as part of systemic disease or as the primary site of involvement is rare. We report a single case of primary (isolated) nodal Churg-Strauss syndrome occurring in an 11-year-old boy with asthma, fever, night sweats, and cervical adenopathy. The clinical diagnosis was lymphoma. The unusual presentation of Churg-Strauss syndrome limited to lymph nodes is important to recognize and diagnose correctly because the administration of steroid therapy is associated with a favorable outcome. PMID- 11037865 TI - Left brachial artery thrombus, left axillary vein thrombus, and stroke in a neonate with factor V Leiden mutation. AB - Factor V Leiden mutation is reportedly the most common hereditary risk factor for thrombosis. Spontaneous venous thromboses in children with factor V Leiden are rare without the presence of an additional risk factor for thrombosis. Spontaneous arterial thromboses are even more rare. In this case report, we describe an unusual case of a neonate born with both arterial and venous thromboses involving the left brachial artery, the left brachial vein, and stroke involving the right middle cerebral artery. The infant was subsequently found to be heterozygous for the factor V Leiden mutation. His mother was also heterozygous for the mutation but did not have a history of thrombosis. Evaluation for the factor V Leiden mutation should form part of the work-up of an infant with either arterial or venous thromboses or stroke because it increases the infant's relative risk for future thrombosis. PMID- 11037866 TI - Primary intestinal non-Hodgkin lymphoma. PMID- 11037867 TI - Diamond-Blackfan anemia and midline defects. PMID- 11037868 TI - Malignant fibrous histiocytoma preceding medulloblastoma. PMID- 11037869 TI - An uncommon cause of dyspnea in a teenage girl: massive pulmonary embolism. PMID- 11037870 TI - Hemolytic uremic syndrome secondary to the treatment of acute lymphoblastic leukemia. PMID- 11037871 TI - A further case of chromosome 8q rearrangement in lipoblastoma. PMID- 11037872 TI - Low-dosage immunoglobulins for an infant with hypogammaglobulinemia, maple syrup urine disease, and parvovirus B19-associated aplastic crisis. PMID- 11037873 TI - Cellular mechanisms and the role of cytokines in bone erosions in rheumatoid arthritis. PMID- 11037874 TI - Acidic fibroblast growth factor in synovial cells. AB - OBJECTIVE: To characterize the production and regulation of acidic fibroblast growth factor (aFGF) in type B (fibroblast-like) synoviocytes cultured from both inflammatory and noninflammatory synovial lesions. METHODS: Immunohistochemistry, Western blotting, and reverse transcriptase-polymerase chain reaction were used to examine the expression of aFGF by synovial cells in vitro. Incorporation of 3H thymidine by NIH3T3 cells in the presence or absence of neutralizing antibody to aFGF was used to measure bioactive aFGF levels in culture media. RESULTS: Acidic FGF was detected in all synovial cell lines during growth in vitro; however, synoviocytes from rheumatoid arthritis (RA) patients sustained more abundant production of cytoplasmic and nuclear aFGF. Acidic FGF production persisted after multiple passages and did not depend on the presence of serum. Both RA and noninflammatory synovial cells were competent to release aFGF into the media, even though aFGF lacks a signal peptide. Tumor necrosis factor alpha, interleukin 6, and epidermal growth factor did not increase aFGF expression in vitro; in contrast, transforming growth factor beta1 (TGFbeta1) was found to markedly increase aFGF production by cultured synovial cells. CONCLUSION: Acidic FGF synthesis and release is a component of synovial cell growth that is markedly increased in RA. TGFbeta1, and not proinflammatory cytokines, is a potent inducer of aFGF production by synoviocytes in vitro. These findings suggest that in RA, interactions between TGFbeta1 and aFGF may contribute to angiogenesis and fibroblast proliferation, potentially independently of inflammatory mediators. PMID- 11037875 TI - Antigen-presenting cells containing bacterial peptidoglycan in synovial tissues of rheumatoid arthritis patients coexpress costimulatory molecules and cytokines. AB - OBJECTIVE: Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by intimal lining hyperplasia and massive infiltration of the synovial sublining by antigen-presenting cells (APCs), lymphocytes, and plasma cells. Peptidoglycan (PG), a major cell wall component of gram-positive bacteria, which is abundantly expressed in all mucosa, is believed to be involved in the pathogenesis of RA because of its ability to induce the production of proinflammatory cytokines as well as to induce arthritis in rodents. While PG has been detected in APCs in RA joints, little is known about the role of these cells in RA. In this study, the presence and immune competence of PG-containing cells in synovial tissues from 14 RA and 14 osteoarthritis (OA) patients were analyzed in situ. METHODS: Using immunohistochemistry, we examined the coexpression of phenotypic markers, costimulatory molecules, and cytokines by PG-containing cells. RESULTS: PG was present in higher numbers in RA than in OA synovial tissues, although the difference was not significant. PG-containing cells were mainly macrophages, but some mature dendritic cells also contained PG. A high percentage of PG-containing cells in both RA and OA synovial tissues coexpressed HLA-DR. CD40, CD80, and CD86 expression by PG-containing cells was higher in RA than in OA tissues. Furthermore, PG-containing cells coexpressed cytokines, which modulate inflammatory reactions, in particular, tumor necrosis factor alpha and interleukins 6 and 10. CONCLUSION: The results suggest that PG-containing cells may contribute to inflammation within the microenvironment of the joint in RA patients. PMID- 11037876 TI - The mechanism of taurine chloramine inhibition of cytokine (interleukin-6, interleukin-8) production by rheumatoid arthritis fibroblast-like synoviocytes. AB - OBJECTIVE: Taurine chloramine (Tau-Cl) has been shown to inhibit the production of proinflammatory cytokines (interleukin-6 [IL-6] and IL-8) by fibroblast-like synoviocytes (FLS) isolated from rheumatoid arthritis (RA) patients. The present study was conducted to elucidate the mechanism of inhibitory action exerted by Tau-Cl. METHODS: The effects of Tau-Cl on 1) the transcription of genes coding for IL-6 and IL-8, and 2) the activity of nuclear factor kappaB (NF-kappaB) and activator protein 1 (AP-1) transcription factors, which are crucial for the transcription of these cytokine genes, were investigated in FLS isolated from the synovial tissue of RA patients. FLS were cultured in vitro for 3-6 passages and stimulated with recombinant human IL-1beta (1 ng/ml) in the presence of either Tau or Tau-Cl, which were added simultaneously with the stimulus at concentrations of 250 microM or 500 microM. The relative expression of IL-6 and IL-8 messenger RNA (mRNA) was evaluated after 4 hours of stimulation, using competitive reverse transcriptase-polymerase chain reaction. The DNA binding activity of NF-kappaB and AP-1 was examined 30 minutes and 2 hours after cell stimulation, respectively, using electromobility gel shift assay. RESULTS: IL 1beta triggered a significant rise in the activity of transcription factors NF kappaB and AP-1, followed by an elevation of cytokine IL-6 and IL-8 mRNA expression. Tau-Cl, but not Tau, reduced IL-1beta-triggered cytokine mRNA expression, exerting stronger inhibitory activity on the levels of IL-6 than on those of IL-8. Importantly, Tau-Cl also diminished the activity of NF-kappaB and, to a lesser extent, that of AP-1 transcription factor. Neither IL-1beta nor Tau Cl affected the activity of octamer transcription factor 1. CONCLUSION: Tau-Cl inhibition of IL-6 and IL-8 synthesis in FLS from RA patients results from the ability of this compound to diminish the activity of the major transcriptional regulators (NF-kappaB and AP-1), which subsequently reduces the transcription of these cytokine genes. PMID- 11037877 TI - Decreased cellular activity and replicative capacity of osteoblastic cells isolated from the periarticular bone of rheumatoid arthritis patients compared with osteoarthritis patients. AB - OBJECTIVE: Periarticular osteopenia is frequently observed in rheumatoid arthritis (RA). Bone loss has been considered to be at least partly due to inadequate bone formation, which in turn, is largely dependent on the number of osteoblasts and the osteoblastic activity. Normal human somatic cells undergo a finite number of cell divisions and ultimately enter a nondividing state called replicative senescence. It has been proposed that the telomere, the terminal sequence of chromosomes, is the mitotic clock that triggers senescence. In the present study, we sought to clarify the relationship between periarticular osteopenia and osteoblast replicative senescence in RA. METHODS: We examined age related changes in cellular activity (alkaline phosphatase activity, osteocalcin and C-terminal type I procollagen secretion, and cAMP response to parathyroid hormone), replicative capacity, and senescent cell expression in osteoblasts from periarticular bone samples obtained from 15 patients with RA and 15 age-matched patients with osteoarthritis (OA). Cellular replicative capacity was analyzed by the mean telomere length and in vitro remaining replicative lifespan of the cells. RESULTS: In both OA and RA groups, the cell proliferation rate, the levels of osteoblastic markers, mean telomere length, and replicative lifespan in osteoblastic cells gradually decreased with the increasing age of the donor. The percentage of senescent osteoblastic cells in the periarticular bone increased with age in both groups, and the rate of expression of senescent cells was higher in RA patients than in age-matched OA patients. The osteoblastic activities and replicative capacity of osteoblastic cells from RA patients were lower than those from OA patients at any donor age. The age-related decreases in the osteoblastic activity and replicative capacity of osteoblastic cells from periarticular bone were greater in RA patients than in OA patients. CONCLUSION: Our results suggest that osteoblast replicative senescence in periarticular bones occurs more rapidly with aging in RA than in OA patients and contributes to periarticular osteopenia in RA. PMID- 11037878 TI - Immortalized human adult articular chondrocytes maintain cartilage-specific phenotype and responses to interleukin-1beta. AB - OBJECTIVE: To develop a reproducible immortalized human chondrocyte culture model for studying the regulation of chondrocyte functions relevant to arthritic diseases in adult humans. METHODS: Primary adult articular chondrocytes were immortalized with a retrovirus expressing a temperature-sensitive mutant of SV40 large T antigen (tsTAg). The established tsT/AC62 chondrocyte cell line was examined in monolayer and alginate culture systems. The levels of messenger RNA (mRNA) encoding cartilage matrix proteins and interleukin-1beta (IL-1beta) inducible mRNA were analyzed by reverse transcriptase-polymerase chain reaction. Matrix protein synthesis was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of 35S-sulfate-labeled proteoglycans and Western blotting of type II collagen and aggrecan. Type II collagen (COL2A1)-luciferase reporter gene expression was analyzed by transient transfection. Phosphorylated stress activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), p38 mitogen activated protein kinase (p38 MAPK), and activating transcription factor 2 (ATF 2) were detected by Western blotting. RESULTS: The tsT/AC62 cells expressed TAg at the permissive temperature (32degrees C), and the loss of TAg at 37 degrees C and 39 degrees C correlated with decreased cell proliferation. Cells in alginate culture deposited abundant alcian blue-stainable matrix and continued to proliferate at 32 degrees C. Preferential retention of aggrecan was observed in the cell-associated matrix, while biglycan and decorin were secreted into the medium of monolayer and alginate cultures. The levels of COL2A1 and aggrecan mRNA were increased after transfer from monolayer to alginate culture at 32 degrees C. Treatment with IL-1beta decreased COL2A1 and aggrecan mRNA levels and increased the levels of matrix metalloproteinases 1, 3, and 13 mRNA, as well as those of cyclooxygenase 2, type I collagen, and secretory phospholipase A2 type IIA mRNA, but not those of inducible nitric oxide synthase mRNA. IL-1beta also stimulated phosphorylation of p38 MAPK, SAPK/JNK, and ATF-2. The p38 MAPK-selective inhibitor, SB203580, partially reversed IL-1beta-induced inhibition of COL2A1 mRNA levels and COL2A1-luciferase reporter gene expression. CONCLUSION: The tsT/AC62 cells provide a reproducible model that mimics the adult articular chondrocyte phenotype, particularly in alginate culture, and demonstrates characteristic responses to IL-1beta. These studies also show, for the first time, that p38 MAPK is one of the signals required for IL-1beta-induced inhibition of COL2A1 gene expression. Availability of this model will permit identification of signals that regulate cytokine responses, and will also provide rational strategies for targeting these pathways. PMID- 11037879 TI - The increased swelling and instantaneous deformation of osteoarthritic cartilage is highly correlated with collagen degradation. AB - OBJECTIVE: To provide evidence for the hypothesis that the loss of tensile strength of osteoarthritic (OA) cartilage (resulting in swelling-the hallmark of OA) is due to an impaired collagen network and not to loss or degradation of proteoglycans. METHODS: The amount of degraded collagen molecules, the fixed charge density (FCD) on a dry-weight basis, the degree of swelling in saline, and the instantaneous deformation (ID; a test reflecting the tensile stiffness of the collagen network) were measured in full-depth OA femoral condyle samples. In addition, levels of the crosslink hydroxylysylpyridinoline (HP), the amount of degraded collagen molecules, and the degree of swelling were determined in the 3 zones (surface, middle, and deep) of OA cartilage. We also compared the ID of normal and OA cartilage. RESULTS: In full-depth OA cartilage, a close relationship was found between swelling and ID. Swelling and ID correlated strongly with the amount of degraded collagen molecules, and were not related to FCD. OA cartilage showed the same zonal pattern in HP levels as normal cartilage (i.e., an increase with depth). No relationship was found between collagen crosslinking and swelling of the surface, middle, and deep zones. In all 3 zones, swelling was proportional to the amount of degraded collagen molecules. Compared with that of normal cartilage, the change in ID of OA cartilage was most pronounced at the surface in a direction parallel to the direction of the collagen fibrils. CONCLUSION: The decreased stiffness of the OA collagen network (as measured by swelling and ID) is strongly related to the amount of degraded collagen molecules. The anisotropy in ID parallel and perpendicular to the direction of the fibrils revealed that the impairment of strength resides mainly in, and not between, the fibrils. Proteoglycans play only a minor role in the degeneration of the tensile stiffness of OA cartilage. PMID- 11037880 TI - Calcium pentosan polysulfate inhibits the catabolism of aggrecan in articular cartilage explant cultures. AB - OBJECTIVE: The catabolism of aggrecan and loss of aggrecan fragments from articular cartilage is a key event in the pathogenesis of arthritic diseases such as osteoarthritis. The catabolism of aggrecan is mediated by the specific proteolytic activity termed aggrecanase. The aim of this study was to investigate the effect of the chondroprotective agent calcium pentosan polysulfate (CaPPS) on the aggrecanase-mediated catabolism of aggrecan. METHODS: The catabolism of 35S labeled aggrecan and loss of tissue glycosaminoglycans (GAGs) were investigated using bovine articular cartilage explant cultures maintained in medium containing varying concentrations of CaPPS (1-100 microg/ml) in the presence or absence of 10(-6)M retinoic acid or 7 ng/ml recombinant human interleukin-1alpha (rHuIL 1alpha). In addition, the effect of CaPPS on the degradation of aggrecan monomers by aggrecanase activity present in conditioned medium from joint capsule explant cultures was investigated. RESULTS: CaPPS inhibited the catabolism of 35S-labeled aggrecan in a dose-dependent manner, particularly when retinoic acid or rHuIL 1alpha was used to stimulate aggrecan catabolism. These effects were reflected in the tissue levels of GAG remaining in these cultures at the end of the experiment. CaPPS inhibited the degradation of aggrecan monomers by soluble aggrecanase activity. CONCLUSION: CaPPS inhibits the catabolism of aggrecan by articular cartilage in a dose-dependent manner, particularly when the processes responsible for aggrecan loss are stimulated. This effect occurs, at least in part, through direct inhibition of aggrecanase activity. CaPPS did not adversely affect overall chondrocyte metabolism, as shown by the incorporation of 35S sulfate and 3H-leucine into macromolecules and by lactate production in cartilage explant cultures. PMID- 11037881 TI - CCAAT binding transcription factor binds and regulates human COL1A1 promoter activity in human dermal fibroblasts: demonstration of increased binding in systemic sclerosis fibroblasts. AB - OBJECTIVE: To determine the binding factors that interact with the proximal promoter region of the human type I collagen gene, COL1A1, and to examine their involvement in its transcriptional regulation in normal and systemic sclerosis (SSc) dermal fibroblasts. METHODS: Nuclear extracts from dermal fibroblasts from 4 patients with SSc and 4 age- and sex-matched control individuals were examined by electrophoresis mobility shift assays with a COL1A1 promoter fragment encompassing nucleotides -174 to -50 bp. Supershift assays with antibodies specific to various transcription factors, and competition experiments using consensus, wild-type, or mutated oligonucleotides corresponding to their specific binding sites, were performed. The effects of specific oligonucleotides as "intracellular competitors" were examined by transient transfection experiments in SSc fibroblasts using a COL1A1 construct containing -174 bp of the promoter. RESULTS: The findings demonstrate that the CCAAT binding transcription factor (CBF) binds the proximal CCAAT box located at -100 to -96 bp, but not the distal CCAAT box at -125 to -121 bp, of the human COL1A1 promoter in both SSc and normal fibroblasts. CBF binding activity was 3-5-fold higher in the SSc fibroblasts. Moreover, the promoter activity of the -174-bp COL1A1 construct was decreased by up to 50% when specific oligonucleotides were used as "intracellular competitors." In addition, Sp1 and Sp3 were other transcription factors found to be involved in the formation of the DNA-protein complexes within this region of the COL1A1 promoter. CONCLUSION: These results indicate that the transcription factor CBF binds the human COL1A1 proximal promoter region in human dermal fibroblasts, and its binding activity is higher in SSc fibroblasts. PMID- 11037882 TI - Role of apoptosis and transforming growth factor beta1 in fibroblast selection and activation in systemic sclerosis. AB - OBJECTIVE: We hypothesized that pathophysiologic events during the development of systemic sclerosis (SSc) may lead to selection and propagation of certain apoptosis-resistant fibroblast subpopulations. The aim of this study was to examine a possible role for apoptosis in fibroblast selection in SSc and the role of transforming growth factor beta1 (TGFbeta1). METHODS: We compared SSc and normal fibroblasts for their susceptibility to anti-Fas-induced apoptosis and analyzed 2 models that might lead to fibroblast resistance to apoptosis in this process: long-term exposure to either anti-Fas or TGFbeta1. RESULTS: SSc-derived fibroblasts were resistant to anti-Fas-induced apoptosis, showing 5.5 +/- 17.2% (mean +/- SD) apoptosis, compared with 32.1 +/- 14.0% among normal fibroblasts (P < 0.05). Anti-Fas-selected normal fibroblasts showed 9.0 +/- 3.7% apoptosis, compared with 21.6 +/- 5.9% for sham-treated cells, which is consistent with the elimination of apoptosis-susceptible subpopulations. Normal fibroblasts subjected to 6 weeks of TGFbeta1 treatment showed not only resistance to apoptosis, but also proliferation (118.5 +/- 35.4%), after anti-Fas treatment, compared with sham-treated cells (35.1 +/- 11.1% apoptotic cell death). TGFbeta1 treatment also increased the proportion of myofibroblasts (47% versus 28% in controls). Cultured SSc fibroblasts had a greater proportion of myofibroblasts (32-83%) than did normal fibroblasts (4-25%). We also examined the relationship between collagen gene expression and the myofibroblast phenotype in normal and SSc skin sections. Only 2 of 7 normal sections had alpha-smooth muscle actin (a-SMA)-positive cells (mean +/- SD score 0.29 +/- 0.49 on a scale of 0-3), but all SSc sections were positive for alpha-SMA, with a mean score of 1.90 +/- 0.88 for lesional and 1.50 +/- 0.71 for nonlesional sections. Scores for alpha1(I) procollagen messenger RNA (mRNA) in lesional skin (mean +/- SD 3.30 +/- 0.82 on a scale of 1-4) were significantly higher than in normal (1.43 +/- 0.79) or nonlesional (1.40 +/- 0.52) skin, but scores varied, and there was no correlation between collagen mRNA and alpha-SMA levels. CONCLUSION: Our results show that resistance to apoptosis is an important part of the SSc phenotype. TGFbeta1 may play a role by inducing apoptosis-resistant fibroblast populations, and also by inducing myofibroblasts and by enhancing extracellular matrix synthesis. PMID- 11037883 TI - Increased phosphorylation of transcription factor Sp1 in scleroderma fibroblasts: association with increased expression of the type I collagen gene. AB - OBJECTIVE: To determine the potential roles of transcription factors Sp1 and Sp3 in the increased expression of the human alpha2(I) collagen gene in scleroderma fibroblasts. METHODS: Dermal fibroblasts from 7 patients with diffuse systemic sclerosis (SSc; scleroderma) of recent onset and from 7 healthy individuals were studied. The levels of expression of alpha2(I) procollagen, Sp1, and Sp3 messenger RNA (mRNA), with or without stimulation by transforming growth factor beta (TGFbeta) or oncostatin M (OSM), were evaluated by Northern blot analysis, and the respective protein levels were determined by immunoblotting. The DNA binding activity of nuclear proteins recognizing the cis-acting elements in the human alpha2(I) collagen promoter was examined by gel mobility shift assays. The levels of Sp1 phosphorylation were investigated by immunoprecipitation using an antiphosphoserine-specific antibody. RESULTS: SSc fibroblasts showed basal alpha2(I) collagen mRNA levels that were approximately 3 times higher than those in normal fibroblasts. TGFbeta or OSM increased human alpha2(I) collagen mRNA expression in normal dermal fibroblasts, but these cytokines failed to increase alpha2(I) collagen mRNA levels in SSc fibroblasts. There were no significant differences in the levels of expression of Sp1 or Sp3 between SSc and normal fibroblasts. However, increased Sp1 phosphorylation was detected in SSc fibroblasts compared with normal fibroblasts. Mithramycin, a specific inhibitor of Sp1 binding, abolished the increased expression of the alpha2(I) collagen gene in SSc fibroblasts, in a dose-dependent manner. CONCLUSION: These results demonstrate the involvement of Sp1 in the up-regulation of expression of the alpha2(I) collagen gene in SSc fibroblasts. PMID- 11037884 TI - Intercellular adhesion molecule 1 and beta2 integrins in C1q-stimulated superoxide production by human neutrophils: an example of a general regulatory mechanism governing acute inflammation. AB - OBJECTIVE: To investigate the role of intercellular adhesion molecule 1 (ICAM-1) and beta2 integrins in the production of superoxide (O2-) by C1q-stimulated human polymorphonuclear leukocytes (PMN). METHODS: PMN were pretreated with F(ab')2 fragments of monoclonal antibodies (mAb) that blocked or did not block beta2 integrin-mediated adhesion. The cells were added to wells coated with C1q, and the production of O2- was monitored kinetically as a color change due to reduction of cytochrome c. In some experiments, C1q was co-immobilized with purified ICAM-1. RESULTS: Blocking mAb to the shared beta2 integrin subunit, CD18, completely inhibited the O2- response triggered by immobilized C1q, while blocking mAb to the alpha subunits of the beta2 integrins each partially blocked the O2- response. PMN treated with C1q were found to activate the beta2 integrins lymphocyte function-associated antigen 1 and CR3 for binding to ICAM-1. Co immobilization of ICAM-1 with C1q cooperatively triggered O2- production by PMN. CONCLUSION: beta2 integrin binding to an ICAM provided an essential costimulatory signal for O2-production triggered by C1q in PMN. Our findings suggest a model for PMN activation in which 2 stimuli are required for O2- production: a first signal that also activates PMN beta2 integrins, followed by a second, beta2 integrin-mediated signal, which occurs physiologically upon PMN binding to ICAM 1. The requirement for this dual signal for PMN generation of O2- would serve as a regulatory mechanism to limit the production of O2- to a tissue environment where C1q, or some other stimulus, is colocalized with stromal cells bearing up regulated ICAM-1. This mechanism may explain why all tissues can express ICAM-1 and may explain in part why inhibitors of tumor necrosis factor alpha, a major physiologic stimulus of ICAM-1 up-regulation, are potent antiinflammatory agents. PMID- 11037885 TI - Acetylcholine prevents intercellular adhesion molecule 1 (CD54)-induced focal adhesion complex assembly in endothelial cells via a nitric oxide-cGMP-dependent pathway. AB - OBJECTIVE: Nitric oxide (NO) is induced by exposure of endothelial cells (EC) to acetylcholine, where it acts in a paracrine manner to relax smooth muscle and as a defensive molecule to inhibit the adhesion of leukocytes to EC. The mechanism(s) of the antiadhesive properties of constitutive NO are poorly understood. In these studies, we found that NO induced by acetylcholine exerts autocrine effects, which interfere with normal adhesion mechanisms. METHODS: The function of the adhesion molecule intercellular adhesion molecule 1 (CD54) of EC was measured using latex beads coated with antibody to CD54 as a model for CD54 ligation by the leukocyte beta2 integrin. Recruitment of filamentous actin (F actin) and of the signaling molecule vasodilator-stimulated phosphoprotein (VASP) was measured by immunofluorescence microscopy. RESULTS: Exposure of EC to anti CD54 beads induced the subplasmalemmal assembly of F-actin and VASP. Acetylcholine blocked the anti-CD54 bead-induced translocation of F-actin and VASP; this effect was reversed by inhibition of NO production. The NO action did not interfere with binding, but completely inhibited the assembly of the focal activation complex, which we believe is necessary for firm heterotypic adhesion between leukocyte and EC. Further studies indicated that the NO effect was due to its capacity to raise cGMP. Platelet endothelial cell adhesion molecule 1 (CD31, also implicated in leukocyte adhesion) did not mimic CD54 responses. CONCLUSION: These results indicate that the ligation of endothelial cell CD54 induces the assembly of subplasmalemmal F-actin and the recruitment of VASP. NO derived from constitutive nitric oxide synthase acts to disrupt these CD54-elicited endothelial cell responses. This action may protect vascular endothelium from leukocyte-mediated injury. PMID- 11037886 TI - Clearance of anti-double-stranded DNA antibodies: the natural immune complex clearance mechanism. AB - OBJECTIVE: To develop an in vitro model for investigating the mechanism by which autoantibodies in immune complexes (ICs) that are bound to primate erythrocytes via antigen-based heteropolymers (AHPs) are cleared from the circulation and localized to the liver. METHODS: IgG anti-double-stranded DNA (anti-dsDNA) antibodies in ICs with dsDNA were bound to human erythrocytes via complement receptor 1 (CR1) either by opsonization with normal human serum as a complement source or through the use of an AHP, which consists of an anti-CR1 monoclonal antibody (mAb) that is chemically crosslinked with dsDNA. We performed parallel investigations of the mechanism of transfer of both types of erythrocyte-bound ICs to a monocytic cell line (U937). Erythrocytes with CR1-bound ICs were incubated with U937 cells under a variety of conditions, and subsequently, the levels of IgG anti-dsDNA, CR1, AHP, or C3b on both erythrocytes and U937 cells were measured by flow cytometry with appropriate fluorescently labeled probes. RESULTS: In the presence of U937 cells, both the AHP-anti-dsDNA and C3b-opsonized ICs were rapidly removed from the erythrocytes; at 37 degrees C, more than half of the complexes were removed in 2 minutes. Monomeric mouse IgG2a mAb blocked the transfer of both types of complexes by 75%, suggesting that Fcgamma receptor type I (FcgammaRI) is the main phagocyte receptor responsible for the removal of ICs from erythrocytes. Levels of CR1 on the erythrocyte surface were reduced during transfer of the AHP-anti-dsDNA ICs, suggesting that transfer involves a concomitant removal of CR1, presumably by proteolysis. CONCLUSION: Transfer of AHP-anti-dsDNA ICs from erythrocyte CR1 to model phagocytes occurs by a mechanism that is similar to the natural mechanism of IC clearance, involving recognition by FcgammaRI and removal of erythrocyte CR1 as key steps. PMID- 11037887 TI - Role of macrophages in Staphylococcus aureus-induced arthritis and sepsis. AB - OBJECTIVE: A model of hematogenously induced Staphylococcus aureus arthritis was used to analyze the role of macrophages in this highly destructive condition. In this model, the majority of cells in the cartilage-synovial junction that participate in the destructive process are macrophages. METHODS: To assess the role of monocytes/macrophages in staphylococcal arthritis, mice were inoculated with S aureus or given phosphate buffered saline as control. Mice were rendered monocytopenic by administration of etoposide, a drug that selectively depletes the monocyte/macrophage population. RESULTS: Throughout the course of infection, the etoposide-treated mice exhibited a significantly less severe arthritis than the control animals. These data were confirmed by histopathologic analysis of the joints. The down-regulation of development of arthritis was accompanied by decreased serum levels of the proinflammatory cytokines tumor necrosis factor alpha and interleukin-6. In contrast, infection-triggered mortality was increased in the etoposide-treated mice as compared with the control animals. Notably, the monocytopenic mice exhibited elevated bacterial burden in the blood and kidneys on days 3 and 7 after inoculation with staphylococci. CONCLUSION: This study indicates a dual role of mononuclear phagocytes in the pathogenesis of S aureus induced infection. On the one hand, absence of macrophages leads to a favorable outcome concerning the severity of arthritic lesions, but on the other hand, the clearance of bacteria by monocytes/macrophages is decreased, resulting in poor survival. PMID- 11037888 TI - The major role of macrophages and their product tumor necrosis factor alpha in the induction of arthritis triggered by bacterial DNA containing CpG motifs. AB - OBJECTIVE: To understand the mechanisms of arthritis triggered by CpG-containing oligonucleotides (ODN). METHODS: Following the induction of CpG ODN-triggered arthritis in mice, we analyzed the impact of depletion of immune cells, including neutrophils, natural killer (NK) cells, and monocyte/macrophages, on the arthritis, as well as the impact in SCID mice lacking T and B cells. In addition, tumor necrosis factor alpha (TNFalpha) knockout mice were studied, and intraarticular administration of p65 antisense to nuclear factor kappaB (NF kappaB) was used to examine effects in CpG ODN-triggered arthritis. Cytokine messenger RNA expression in synovial tissue was evaluated by in situ hybridization. RESULTS: The presence of macrophages was mandatory for the mediation of arthritis triggered by CpG ODN, whereas the absence of neutrophils, NK cells, T cells, and B cells was of minor importance in this regard. The proinflammatory cytokines TNFalpha, interleukin-1beta, and interleukin-12, which originate from macrophages, were frequently found in the inflamed joints, and TNFalpha was confirmed to be an important mediator in the development of arthritis, since the incidence and severity of joint inflammation were markedly reduced in TNFalpha knockout mice. NF-kappaB exerted an important regulatory role in the development of CpG ODN-mediated arthritis, since local administration of antisense to the p65 subunit of NF-kappaB diminished the incidence of inflammation by 50%. CONCLUSION: Macrophages and their products play an important role in the development of arthritis triggered by bacterial DNA containing CpG motifs. PMID- 11037889 TI - Development of spontaneous arthritis in beta2-microglobulin-deficient mice without expression of HLA-B27: association with deficiency of endogenous major histocompatibility complex class I expression. AB - OBJECTIVE: Mice deficient in beta2-microglobulin (beta2m), but expressing the human major histocompatibility complex (MHC) class I molecule HLA-B27, have been reported to develop spontaneous inflammatory arthritis (SA). We sought to determine whether, under certain conditions, beta2m deficiency alone was sufficient to cause SA, and if this might be a result of class I deficiency. METHODS: The following types of mice were produced: mice of the MHC b haplotype genetically deficient in beta2m (beta2m(0)) on several genetic backgrounds (C57BL/6J [B6], BALB/cJ, SJL/J, MRL/MpJ, and B6,129), mice deficient in the transporter associated with antigen processing (TAP1(0)) on a B6,129 background, and HLA-B27-transgenic beta2m(0) mice on a B6 background. Cohorts were transferred from specific pathogen-free (SPF) to conventional (non-SPF) animal rooms, and evaluated clinically and histologically for the development of SA. RESULTS: SA occurred in TAP1(0) and beta2m(0)/class I-deficient mice with a mixed B6,129 genome at a frequency of 30-50%, while 10-15% of B6, SJL/J, and BALB/cJ beta2m(0) mice developed this arthropathy. MRL/ MpJ beta2m(0) mice were unaffected. Expression of B27 did not increase the frequency of SA in B27 transgenic B2m(0) B6 mice compared with that in beta2m(0) B6 controls. CONCLUSION: Class I deficiency is sufficient to cause SA in mice. The frequency of disease, as well as B27-specific SA, is markedly dependent on a non-MHC genetic background. These results suggest that class I deficiency in a genetically susceptible mouse can mimic B27-associated arthropathy. PMID- 11037890 TI - Evidence for antimuscarinic acetylcholine receptor antibody-mediated secretory dysfunction in nod mice. AB - OBJECTIVE: Antibodies directed against general and specific target-organ autoantigens are present in the sera of human patients and animal models with autoimmune disease. The relevance of these autoantibodies to the disease process remains ambiguous in most cases. In autoimmune exocrinopathy (Sjogren's syndrome), autoantibodies to the intracellular nuclear proteins SSA/Ro and SSB/La, as well as the cell surface muscarinic cholinergic receptor (M3) are observed. To evaluate the potential role of these factors in the loss of secretory function of exocrine tissues, a panel of monoclonal and polyclonal antibodies was developed for passive transfer into the NOD animal model. METHODS: Monoclonal antibodies to mouse SSB/La, rat M3 receptor, and a rabbit polyclonal antiparotid secretory protein antibody were obtained for this study. These antibody reagents were subsequently infused into NOD-scid mice. Saliva flow rates were subsequently monitored over a 72-hour period. Submandibular gland lysates were examined by Western blotting for alteration of the distribution of the water channel protein aquaporin (AQP). RESULTS: Evaluation of the secretory response indicated that only antibodies directed toward the extracellular domains of the M3 receptor were capable of mediating the exocrine dysfunction aspect of the clinical pathology of the autoimmune disease. In vitro stimulation with a muscarinic agonist of submandibular gland cells isolated from mice treated with anti-M3 antibody, but not saline or the isotype control, failed to translocate AQP to the plasma membrane. CONCLUSION: These findings define a clear role for the humoral immune response and the targeting of the cell surface M3 signal transduction receptor as primary events in the development of clinical symptoms of autoimmune exocrinopathy. Furthermore, the anti-M3 receptor activity may negatively affect the secretory response through perturbation of normal signal transduction events, leading to translocation of the epithelial cell water channel. PMID- 11037891 TI - Nucleosomes are major T and B cell autoantigens in systemic lupus erythematosus. AB - OBJECTIVE: Double-stranded DNA (dsDNA) is a well-known target of autoantibodies in systemic lupus erythematosus (SLE). The majority of these autoantibodies are of the IgG isotype and show affinity maturation, both of which are known hallmarks of T cell help. T cell responses to autoantigens, including DNA, have been reported only incidentally in SLE patients. Nevertheless, in murine SLE, naked DNA and complexed DNA (nucleosomes) are known to be recognized by T cells. This study aimed to characterize the antinucleosome response and its clinical impact on human SLE. METHODS: Nucleosomes were prepared from chicken erythrocytes. Sera from SLE and control patients were investigated by enzyme linked immunosorbent assay (ELISA) for nucleosome-specific antibody responses. Peripheral blood mononuclear cells (PBMC) from SLE and control patients were analyzed by a kinetic T cell proliferation assay. PBMC were subsequently analyzed for nucleosome-specific T cell proliferation. RESULTS: Of 136 SLE patients, 56% were seropositive for antinucleosome antibodies. In contrast, only 3% of 309 control patients (with rheumatoid arthritis, mixed connective tissue disease, undifferentiated connective tissue disease, Lyme borreliosis, scleroderma, Sjogren's syndrome, ulcerative colitis, hepatitis B virus infection, or human immunodeficiency virus infection) were seropositive. Thus, the antinucleosome ELISA had a sensitivity of 56%, a specificity of 97%, and a diagnostic confidence of 90% when applied to SLE. It was therefore superior to an anti-DNA ELISA that demonstrated a 69% diagnostic confidence in the same population. Antinucleosome reactivity in SLE patients correlated significantly with disease activity (P < 0.0001), nephritis (P < 0.002), and psychosis (P < 0.02). When proliferation assays were applied, 14 of 26 SLE patients (54%) were positive for nucleosome specific T cells that proliferated in response to their cognate antigen. A suppressed response was elicited in 3 SLE patients (12%); in these patients, the PBMC response to nucleosomes was lower than the proliferation of PBMC in the presence of culture medium only. PBMC from the remaining 9 SLE patients (35%) were nonresponsive to nucleosomes in either way. Responding, nonresponding, and suppressed populations differed from each other significantly (P < 0.0001). None of the PBMC from 7 healthy donors and 10 control patients could be stimulated with nucleosomal antigens. CONCLUSION: We present evidence that nucleosomes are major autoantigens in human SLE and that antinucleosomal antibodies are highly specific for the disease. The antinucleosome ELISA has been shown to be superior to the anti-dsDNA ELISA and may thus be a significantly better tool for diagnosing SLE. Nucleosome-specific T cells in SLE patients may help B cells class switch to IgG and undergo affinity maturation. PMID- 11037892 TI - A cost-effectiveness analysis of treatment options for patients with methotrexate resistant rheumatoid arthritis. AB - OBJECTIVE: Recently, new treatment options for rheumatoid arthritis (RA) patients with an inadequate response to methotrexate (MTX) have become available. Given the wide variability in efficacy and costs among these different treatment options, we sought to determine their cost-effectiveness (CE) in order to guide policy in different cost-constrained settings. METHODS: We performed a CE analysis comparing 6 treatment options for patients with MTX-resistant RA: 1) etanercept + MTX, 2) etanercept monotherapy, 3) cyclosporine + MTX, 4) triple therapy (hydroxychloroquine, sulfasalazine, and MTX), 5) continuation of MTX monotherapy, and 6) no second-line agent. A decision model was used with a time horizon of 6 months. We used 2 measures of effectiveness based on published clinical trial data: the American College of Rheumatology 20% response criteria (ACR 20); and a weighted average of proportions of patients achieving responses of ACR 70, ACR 50, and ACR 20 (ACR 70 weighted response [ACR 70WR]). Incremental CE ratios were calculated as the additional cost per patient achieving either outcome, compared with the next least expensive option. To help interpret CE relative to these RA-specific outcomes, we conducted a separate, "reference" CE analysis of MTX use in MTX-naive RA patients, using the same outcomes. RESULTS: In our reference analysis, MTX therapy for MTX-naive RA cost $1,100 per ACR 20 outcome and $1,500 per ACR 70WR, compared with no second-line agent. In our base case analysis with either outcome, MTX continuation, cyclosporine + MTX, and etanercept monotherapy cost more, but either were not more efficacious or had a higher incremental CE ratio than the next most expensive option (i.e., they were dominated). Therefore, these options were not cost-effective. The least expensive option, triple therapy, cost 1.3 times more per patient with ACR 20 outcome ($1,500/ACR 20) and 2.1 times more per ACR 70WR ($3,100/ACR 70WR) than MTX therapy for MTX-naive RA. The most efficacious option, the combination of etanercept and MTX, cost 38 times more per patient with ACR 20 outcome ($4,600/ACR 20) and 23 times more per ACR 70WR ($34,800/ACR 70WR) than MTX therapy for MTX-naive RA. Overall, the results of extensive sensitivity analyses did not substantially affect these results. CONCLUSION: Our analysis indicates that if 15 mg/week MTX is cost-effective for achieving ACR 20 or ACR 70WR in MTX naive RA over a 6-month period, then most likely so is triple therapy in MTX resistant RA. Whether etanercept + MTX is cost-effective depends on whether $34,800/ACR 70WR (or $42,600/ACR 20) over a 6-month period is considered acceptable. PMID- 11037893 TI - Fcgamma receptor type IIIA is associated with rheumatoid arthritis in two distinct ethnic groups. AB - OBJECTIVE: To investigate a possible association between a functional polymorphism in the intermediate-affinity receptor for IgG called Fc-gamma receptor type IIIA (FcgammaRIIIA [CD16]) and rheumatoid arthritis (RA). METHODS: This was an allelic association study in which a single nucleotide polymorphism in FcgammaRIIIA was examined as a susceptibility and/or severity factor for RA. The FcgammaRIIIA-158V/F polymorphism was genotyped by direct sequencing in 2 well characterized ethnic groups, UK Caucasians (141 RA patients and 124 controls) and North Indians and Pakistanis (108 RA patients and 113 controls). RESULTS: The FcgammaRIIIA-158V/F polymorphism was associated with RA in both ethnic groups (P = 0.028 for UK Caucasians, P = 0.050 for North Indians and Pakistanis, and P = 0.003 for both groups combined). FcgammaRIIIA-158VF and -158W individuals had an increased risk of developing RA in both populations (UK Caucasians odds ratio [OR] 1.6, P = 0.050; North Indians and Pakistanis OR 1.9, P = 0.023; and combined groups OR 1.7, P = 0.003). In the UK Caucasian group, the highest risk was for nodular RA, a more severe disease subset, associated with homozygosity for the FcgammaRIIIA-158V allele (OR 4.4, P = 0.004). There was also evidence for an interaction between the RA-associated HLA-DRB1 allele and the presence of at least 1 FcgammaRIIIA-158V allele in predicting susceptibility to RA (OR 5.5, P = 0.000). CONCLUSION: We have demonstrated that the FcgammaRIIIA-158V/F polymorphism is a susceptibility and/or severity marker for RA in 2 distinct ethnic groups. This finding may ultimately provide additional insights into the pathogenesis of RA and other autoantibody/immune complex-driven autoimmune diseases. PMID- 11037894 TI - Juvenile rheumatoid arthritis: linkage to HLA demonstrated by allele sharing in affected sibpairs. AB - OBJECTIVE: To test for linkage between the HLA region and juvenile rheumatoid arthritis (JRA), with stratification by onset and course types, in a cohort of affected sibling pairs (ASPs). METHODS: Eighty pairs of siblings with JRA who were registered with the Research Registry for JRA ASPs (sponsored by the National Institute of Arthritis and Musculoskeletal and Skin Diseases) were typed for HLA-DR. The observed ratio of sharing of none, one, or both parental DR alleles was compared against the expected ratio of 1:2:1 by goodness-of-fit chi square tests. A group of 265 unrelated control subjects served as a comparison population for HLA-DR allele frequencies among patients, by Fisher's exact test. RESULTS: Overall, there was excess sharing of 2 DR alleles among ASPs with JRA. The observed ratio of sharing 0, 1, or 2 DR alleles was 8:40:32, instead of the expected ratio of 20:40:20 (P < 0.001). When stratified by JRA onset type, excess allele sharing was demonstrated among ASPs who were concordant for onset type (P = 0.002). This was true for both pauciarticular and polyarticular onset. When stratified by disease course, excess allele sharing was also demonstrated among ASPs who were concordant for disease course (P < 0.001). This was true for both the pauciarticular and the polyarticular course. Among the 32 ASPs who shared two DR alleles, 5 pairs had both DR8 and DR11, which was significantly more frequent (P < 0.0001) than the incidence in the control group (n = 0). CONCLUSION: This study of an independent cohort of multiplex families confirms the previously reported linkage between pauciarticular JRA and the HLA-DR region that was identified using a different analytic method in a cohort of simplex families. Additionally, this study establishes evidence for linkage between polyarticular JRA and the HLA-DR region. PMID- 11037895 TI - Efficacy and safety of diacerein in osteoarthritis of the knee: a double-blind, placebo-controlled trial. The Diacerein Study Group. AB - OBJECTIVE: To evaluate the efficacy and safety of diacerein, a drug with interleukin-1beta--inhibitory activity in vitro, in patients with knee osteoarthritis (OA). METHODS: A total of 484 patients fulfilling the American College of Rheumatology criteria for knee OA were enrolled in this 16-week, randomized, double-blind, placebo-controlled, parallel study group with 3 diacerein dosages of 50 mg/day, 100 mg/day, and 150 mg/day (administered twice daily). RESULTS: In the intent-to-treat population, 100 mg/day diacerein (50 mg twice daily) was significantly superior (P < 0.05) to placebo using the primary criterion (visual analog scale [VAS] assessment of pain on movement). Significant improvement (P < 0.05) was also observed for the secondary criteria, which included the Western Ontario and McMaster Universities OA Index (WOMAC), the WOMAC subscores, and the VAS assessment of handicap. In patients treated with diacerein dosages of 50 mg/day and 150 mg/day, favorable but not significant results were observed for the primary criterion. The best daily dosage of diacerein, calculated from the effect on the VAS assessment of pain on movement, was 90.1 mg. In the per-protocol population, the analysis of the primary criterion showed significant dose-dependent differences (P < 0.05) between each of the 3 diacerein dosages and the placebo. No differences were observed among the 3 diacerein groups. A significantly higher incidence (P < 0.05) of adverse events (AEs), as well as a higher rate of dropoout due to AEs, was observed in patients treated with 150 mg/day diacerein versus those treated with placebo, 50 mg/day diacerein, or 100 mg/day diacerein. Mild-to-moderate transient changes in bowel habits were the most frequent AEs, increasing with the dosage. CONCLUSION: Diacerein, a drug for the treatment of OA, was shown to be an effective treatment for symptoms in patients with knee OA. Taking into account both efficacy and safety, the optimal daily dosage of diacerein for patients with knee OA is 100 mg/day (50 mg twice daily). PMID- 11037896 TI - The 4G/5G polymorphism of the type 1 plasminogen activator inhibitor gene and thrombosis in patients with antiphospholipid syndrome. AB - OBJECTIVE: To investigate the relationship between the 4G/5G polymorphism of the type 1 plasminogen activator inhibitor (PAI-1) gene and thrombotic manifestations in patients with antiphospholipid syndrome (APS). METHODS: We studied a total of 247 patients included in the following 4 groups: 70 patients with primary APS, 104 patients with systemic lupus erythematosus (40 with antiphospholipid antibodies [aPL] and clinical [secondary] APS, 13 with aPL but without clinical APS, and 51 with neither detectable aPL nor a history of thrombosis), 14 asymptomatic individuals with aPL, and 59 patients with thrombosis but without known thrombosis risk factors. A control group of 100 healthy individuals was also analyzed. PAI-1 4G/5G polymorphism was determined by polymerase chain reaction and endonuclease digestion. RESULTS: The allele frequency of 4G/5G in controls was 0.47/0.53. There were no differences in allele distribution among patient groups or between patients and controls. However, a higher frequency of the 4G allele was observed in APS patients with versus those without thrombosis (0.57 versus 0.39; P < 0.05) (odds ratio [OR] 2.83, 95% confidence interval [95% CI] 1.18-6.76). This higher frequency of the 4G allele was attributable to the higher frequency in patients with versus those without arterial thrombosis (0.64 versus 0.43; P < 0.01) (OR 5.96, 95% CI 1.67-21.32), while patients with venous thrombosis had an allele distribution similar to that of those without venous thrombosis (0.49 versus 0.50; P not significant). There was a trend toward higher PAI-1 antigen and activity levels in APS patients and controls with the 4G/4G genotype, but this did not reach statistical significance. CONCLUSION: The presence of the 4G allele of the 4G/5G polymorphism of the PAI-1 gene may be an additional risk factor for the development of arterial thrombosis in APS. PMID- 11037897 TI - Muscle abnormalities in juvenile dermatomyositis patients: P-31 magnetic resonance spectroscopy studies. AB - OBJECTIVE: To characterize metabolic abnormalities in the muscles of children with the juvenile variant of dermatomyositis (JDM) by the use of noninvasive P-31 magnetic resonance spectroscopy (MRS). METHODS: Thirteen patients with JDM (ages 4-16 years) were studied. Biochemical status was evaluated with P-31 MRS by determining the concentrations of the high-energy phosphate compounds, ATP and phosphocreatine (PCr), ratios of inorganic phosphate (Pi) to PCr (Pi:PCr ratio), levels of free cytosolic ADP, and phosphorylation potentials (PPs) during rest, exercise, and recovery. RESULTS: Significant metabolic abnormalities were observed in the thigh muscles of 10 severely affected patients during rest, 2 graded levels of exercise, and recovery. Mean ATP and PCr levels in the muscles of JDM patients were 35-40% below the normal control values (P < 0.003). These data, along with elevated Pi:PCr ratios, higher ADP levels, and abnormal values for PPs, indicated defective oxidative phosphorylation in the mitochondria of diseased JDM muscles. MRS findings were normal in 2 additional patients who had improved with prednisone treatment and in 1 patient who had no muscle weakness (amyopathic variant of JDM). CONCLUSION: JDM patients can be monitored with noninvasive P-31 MRS without sedation. Biochemical defects in energy metabolism are concordant with the weakness and fatigue reported by JDM patients. Quantitative MRS data are useful for evaluating patients and optimizing drug treatment regimens. PMID- 11037898 TI - TNFalpha-308A allele in juvenile dermatomyositis: association with increased production of tumor necrosis factor alpha, disease duration, and pathologic calcifications. AB - OBJECTIVE: To characterize the association between the TNFalpha-308A allele and 1) duration of active disease, 2) peripheral blood mononuclear cell (PBMC) synthesis of tumor necrosis factor alpha (TNFalpha) in vitro, and 3) pathologic calcifications in patients with juvenile dermatomyositis (DM). METHODS: The TNFalpha-308 alleles were determined by polymerase chain reaction in 37 white patients with juvenile DM and in 29 control subjects. Patients were grouped according to duration of immunosuppressive therapy: long (> or =36 months) or short (<36 months). Unstimulated PBMC were examined by enzyme-linked immunosorbent assay for TNFalpha production in vitro. Sixty-five white patients with juvenile DM were examined for pathologic calcifications. RESULTS: TNFalpha 308A was identified in 18 of 37 patients with juvenile DM, in contrast with 5 of 29 controls (P = 0.009). Sixteen of the 18 patients with juvenile DM who had the TNFalpha-308A allele had a disease course > or =36 months, compared with 6 of 19 patients with TNFalpha-308G (P = 0.001). PBMC from 16 of the 18 juvenile DM patients with TNFalpha-308A synthesized more TNFalpha (median 53 pg/ml) compared with PBMC from 9 of 19 patients with TNFalpha-308G (median 19 pg/ml) (P = 0.007). Nineteen of 22 juvenile DM patients requiring therapy for > or =36 months produced more TNFalpha (median 20.5 pg/ml) in comparison with 6 of 15 juvenile DM patients with a <36-month treatment course (median TNFalpha 0.0 pg/ml) (P = 0.005). Detectable calcifications were present in 3 of 8 children with juvenile DM who had TNFalpha-308AA, compared with 2 of 21 children with TNFalpha-308AG and 1 of 36 children who had TNFalpha-308GG (P = 0.017). CONCLUSION: A long course of juvenile DM and the presence of pathologic calcifications were associated with the TNFalpha-308A allele and with the increased production of TNFalpha, which may perpetuate the inflammatory response. PMID- 11037899 TI - HLA-B44031;DRB1*1503 and other sub-Saharan African major histocompatibility complex haplotypes in African Americans and Afro-Caribbeans carry C4A gene deletions: implications for ethnicity-specific lupus susceptibility genes. PMID- 11037900 TI - Adoption by lupus patients who are infertile: comment on the article by Guballa et al. PMID- 11037901 TI - Effects of prostaglandins on form deprivation myopia in the chick. AB - PURPOSE: To investigate the possible role of endogenous prostaglandins in the development of form deprivation myopia, as well as the effects of exogenous prostaglandins using atropine as a positive control. METHODS: Monocular form deprivation was accomplished by mounting a translucent occluder on one eye of 2-3 day old chicks for 1-4 weeks. Ocular occlusion for 1-2 weeks was used for pharmacological blocking experiments. The axial length of the eye was measured by ultrasonography. RESULTS: Indomethacin, administered intramuscularly, subconjunctivally or intravitreally had no significant effect on myopia development. Exogenous PGE2, PGF2alpha and latanoprost acid administered subconjunctivally, or topically as isopropyl ester eyedrops had no statistically significant effect on the myopia development. However, PGF2alpha significantly (p<0.01) attenuated the development of myopia after intravitreal injection. The other two prostaglandins had no statistically significant effect. CONCLUSIONS: Endogenous prostaglandins are unlikely to play a significant role in the development of form deprivation myopia in the chick. However, PGF2alpha suprisingly seems to retard the development of form deprivation myopia, but only when administered intravitreally. Whether the mechanism of the myopia retardation is direct or indirect remains unknown. PMID- 11037902 TI - Myopia profile in Copenhagen medical students 1996-98. Refractive stability over a century is suggested. AB - PURPOSE: To assess the myopia prevalence and profile in today's Danish medical students and to compare the pattern with previous and contemporary investigations. METHODS: As part of the tutoring in ophthalmology 1996-98 Copenhagen medical students of five successive terms (in their 10th semester, n=294) were interviewed and spot-checked about their refraction. Myopia from -0.5 D was recorded and myopia onset age stated. RESULTS: The age range was 22-41 years, median age 26. Myopia ranging from -0.5 to -8 D was encountered in 147 students (50% of all; females 53.9%, males 45%). In seven the myopia was unilateral. Median values for myopia degree and onset age were -2.5 D and 16 years in female myopes (n=89) which differed significantly from the male values (n=58) of -1.5 D and 18 years. Including the age of 18 as lower cut-off for adult onset myopia 44.9% of the myopes belonged to this category. CONCLUSIONS: The results are in close agreement with recent Norwegian student investigations, and there is no obvious trend of a worsening of the academic myopia issue in our part of the world. This is in marked contrast to student myopia profiles as reported in Taiwan, Singapore, and Hong Kong where myopia is booming, by number and degree. PMID- 11037903 TI - Colour contrast sensitivity in cataract and pseudophakia. AB - PURPOSE: To study the influence of cataract on peripheral and central colour contrast sensitivity. METHODS: Peripheral and central colour contrast sensitivity was measured with a computer graphics system along the protan, deutan and tritan axes. Included were 30 patients with cataract divided into three sub-groups: cortical cataract, nuclear sclerosis and posterior subcapsular cataract. Colour contrast was measured before and after cataract operation. RESULTS: There were significant differences in peripheral colour contrast thresholds comparing the preoperative and postoperative results. This difference existed even in patients (n=19) with a pre-operative visual acuity > or = 0.5 (mean 0.6). The tritan axis was the one most affected by cataract. There was no significant difference between cataract sub-groups. Also, the central colour contrast sensitivity was affected by cataract. Again, the tritan axis was the most affected one. There was no significant difference between the cataract sub-groups. We also found large and significant differences in central colour contrast thresholds between normal subjects and postoperative values from the cataract group in all colour axes. The colour contrast sensitivity was poorer in pseudophakes than in normals. There was a difference between the three groups of different IOL material used (PMMA, acrylic and silicone). The difference was significant in the protan axis, the acrylic group having the best colour contrast sensitivity. CONCLUSION: Peripheral colour contrast sensitivity was affected by cataract, even when only moderately developed. This finding is of importance and should be considered when the method is used to study other eye diseases e.g. glaucoma. Central colour contrast sensitivity was also affected by cataract. The pseudophakes were found to have poorer colour contrast sensitivity than normals. The material in the IOL seemed to be of importance for colour contrast. PMID- 11037904 TI - Contrast sensitivity function in patients with beta-thalassemia major. AB - PURPOSE: To investigate contrast sensitivity function in patients with beta thalassemia major, after regular transfusion and chelation therapy. METHODS: We measured contrast sensitivity at four spatial frequencies in 30 patients with beta-thalassemia major and in 30 matched normal control subjects. All subjects underwent an ophthalmic examination that included fluorescein angiography. The contrast sensitivity results from the two groups were compared between them. Patients' contrast sensitivity values were correlated to the variables age, duration of transfusion, duration of chelation therapy and serum ferritin levels, to select the important predictors. RESULTS: Contrast sensitivity function in all beta-thalassemic patients was significantly lower (p<0.0001) compared to the normal control subjects, for all spatial frequencies tested. The most important predictor of contrast sensitivity loss was patients' age. CONCLUSION: Contrast sensitivity testing can detect early changes in the visual function of beta thalassemic patients and should be considered as a monitor for patients under chronic transfusion-chelation therapy. PMID- 11037905 TI - The effect of phenylephrine on pain and flare intensity in eyes with uveitis. AB - PURPOSE: To investigate the influence of protein concentration in the anterior chamber, measured by laser flare meter, on pain sensation after phenylephrine instillation in patients with iridocyclitis. METHODS: Twenty-five consecutive patients with iridocyclitis were included. Patients with cataract, exfoliation syndrome, diabetes mellitus, glaucoma or any other previous ocular diseases or ocular surgery were excluded. Patients were divided into two groups: Group 1- without fibrinoid reaction (FR) in the anterior chamber (18 patients), and Group 2--with FR (7 patients). Protein concentration in the anterior chamber was measured with laser flare meter (FC 500, Kowa Co., Japan). Pupil size was measured by Alcon Tilo Scale, and pain sensation was estimated by Visual Analogue Scale (VAS, Kabi Pharmacia). All measurements were done before and 1 hour after topical instillation of 10% phenylephrine hydrochloride into the subconjunctival sac of the inflamed eyes. RESULTS: Eyes with iridocyclitis and fibrinoid reaction (FR) have a higher flare intensity compared to those without FR (p<0.05). Pupil size was significantly increased after phenylephrine instillation in both study groups (Wilcoxon test, p<0.05). The VAS pain and flare intensity were significantly decreased in group without FR after phenylephrine instillation (Group 1) compared to values before treatment (Wilcoxon test, p<0.05). In eyes with FR (Group 2), no significant influence of phenylephrine instillation was found on VAS pain and flare intensity. CONCLUSIONS: After phenylephrine instillation, flare intensity and pain were significantly decreased only in eyes with iridocyclitis and without FR. The decreasing level of flare intensity, and paralysis of the pupil after phenylephrine instillation seem to alleviate pain in those eyes. PMID- 11037906 TI - Reliability of computerized perimetric threshold tests as assessed by reliability indices and threshold reproducibility in patients with suspect and manifest glaucoma. AB - BACKGROUND: High reproducibility of test measurements is often considered an indication of high reliability of test results. The aim of the current study was to analyse the role of the traditional perimetric reliability indices, False Negative and False Positive responses, and Fixation Losses, as indicators of test reliability in comparison with threshold reproducibility in patients with suspect or manifest glaucoma. METHODS: Perimetry was performed in one eye in each of 76 patients. Each eye was tested twice within approximately one week using the Humphrey II 30-2 SITA STANDARD program. Frequencies of False Positive and False Negative answers, and rates of Fixation Losses were related to threshold reproducibility and to general field status as expressed by Mean Deviation from age-normal threshold values using stepwise multiple linear regression analysis. RESULTS: Substantial field loss was associated both with low threshold reproducibility (p<0.0001) and with increased frequency of False Negative answers (p=0.047). The traditional reliability indices contributed marginally compared to amount of field loss when predicting threshold reproducibility; the coefficient of determination decreased non-significantly from 0.37 to 0.33 when excluding the three reliability indices as explanatory variables from the regression model. Frequencies of False Positive answers and Fixation Losses showed no association to field status or to threshold reproducibility. CONCLUSION: Reliability of visual field test results in patients with glaucoma expressed as threshold reproducibility, can be predicted by amount of field loss alone, and traditional patient reliability indices contribute surprisingly little in this regard. PMID- 11037907 TI - Markers of thrombophilia in retinal vein thrombosis. AB - PURPOSE: To study the prevalence of risk factors for systemic thromboembolism in patients with retinal vein thrombosis (RVT). METHODS: Fifty-four patients younger than 70 years, diagnosed with a retinal vein thrombosis at the Department of Ophthalmology, Aarhus University Hospital, were examined for the presence of venous thrombosis risk factors. RESULTS: 23 patients had a central RVT, 26 had a branch RVT, and 4 had a macular RVT. Nineteen (35.2%) of the patients displayed increased levels of plasma homocysteine, one patient (1.9%) the Factor V Leiden mutation, and one patient (1.9%) displayed an antiphospholipid antibody. All other tests for thrombophilia rendered normal. In 15 of the patients with hyperhomocysteinemia, folic acid substitution returned plasma homocysteine to a normal value in 12 cases. CONCLUSION: A surprisingly high prevalence of hyperhomocysteinemia was detected in this cohort of RVT patients, clearly superseding the prevalence of around 17% found in patients suffering venous thromboembolism in other vascular compartments. Our finding points to the likelihood that hyperhomocysteinemia may be a significant risk factor for retinal vein thrombosis. PMID- 11037908 TI - The anterior lens capsule used as support material in RPE cell-transplantation. AB - PURPOSE: To investigate the use of an ocular basement membrane as support material for transplanted porcine RPE cells. METHODS: Porcine RPE cells were grown on bovine corneal extracellular matrix (ECM), isolated bovine- and porcine lens capsules, and tissue culture plastic. Cell density, and cell morphology were studied by phase contrast microscopy and transmission electron microscopy. RESULTS: RPE cells grown on porcine anterior lens capsule and on ECM obtained better morphology and higher final cell density than cells grown on plastic and on bovine anterior lens capsule. It was possible to transplant the porcine anterior lens capsule to the subretinal space in pigs. Within two weeks of observation, the lens capsule was well tolerated in the subretinal space. CONCLUSION: The anterior lens capsule seems to be promising as support material for use in RPE cell-transplantation. PMID- 11037909 TI - Chronic bilateral keratitis in autoimmune polyendocrinopathy-candidiadis ectodermal dystrophy (APECED). A long-term follow-up and visual prognosis. AB - PURPOSE: To report the outcome of chronic bilateral keratitis and other ocular manifestations in autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED). MATERIAL AND METHODS: The material consists of 69 patients with APECED, who were examined by the senior author as referrals from the pediatricians. The patients were seen at the Helsinki University Eye Hospital a total of 370 times and the follow-up periods range from 2 to 25 years. RESULTS: 25% of the patients had chronic bilateral keratitis with symptoms of intense photophobia, blepharospasm and lacrimation. On 13 patients the first symptoms appeared before the age of 4 years. Keratitis was the first presenting sign before any evidence of systemic disease in three patients, and in all patients keratitis was among the first three manifestations of the syndrome. The clinical picture from the acute to the chronic cicatricial stage is described. The best corrected visual acuity at the end of the follow-up was 0.6 or better in six patients only. Of the total material 12 patients had lenticular opacities as the manifestation of hypoparathyroidism. CONCLUSIONS: Chronic bilateral keratitis is an ocular manifestation of APECED and in its etiology both hypoparathyroidism and candidiasis may be ruled out as single causes. When an ophthalmologist is confronted with a young child with bilateral chronic keratitis with symptoms of intense photophobia, blepharospasm, lacrimation and either with mucocutaneous candidiasis or hypoparathyroidism, the patient should be referred to a pediatric endocrinological consultation for the possibility of underlying APECED. PMID- 11037910 TI - Search for autoantibodies against the HNK-1 carbohydrate epitope in the human eye in intermediate uveitis. AB - PURPOSE: Molecules bearing the immunogenic HNK-1 epitope are present in the inner connective tissue layer and epithelia of the ciliary body. We investigated whether autoantibodies to these molecules can be detected in patients with intermediate uveitis, which affects the ciliary body. METHODS: Serum was collected from 9 patients with intermediate uveitis, and from 6 controls with idiopathic iritis and 3 controls with sarcoid uveitis, representing nongranulomatous and granulomatous uveitis, respectively. The sera were used as polyclonal antibodies to immunostain 3 formalin-fixed, paraffin-embedded normal human donor eyes by the avidin-biotinylated peroxidase complex method. RESULTS: No immunostaining in the ciliary body could be detected using the sera from patients with intermediate uveitis or from the controls. Serum within blood vessels was nonspecifically immunolabelled with the secondary anti-human anti serum. CONCLUSION: No autoantibodies against the HNK-1 epitope or other antigens of the ciliary body could be demonstrated in patients with intermediate uveitis. It is unlikely that such autoantibodies against the HNK-1 epitope have a role in intermediate uveitis. PMID- 11037911 TI - Short term visual prognosis after retinal laser photocoagulation for diabetic maculopathy. AB - PURPOSE: Retinal photocoagulation can improve the visual prognosis of patients with diabetic maculopathy complicated with clinically significant macular oedema. However, this effect covers a wide variation of visual outcome with some patients improving and other patients worsening several visual acuity steps. Therefore, parameters are needed that can be used to ensure that treatment is modified or avoided in those patients who are at risk of experiencing visual loss. METHODS: The change in visual acuity shortly after laser photocoagulation for diabetic maculopathy was assessed in 95 eyes of 79 patients as a part of a routine quality assessment programme, and was compared to the age at onset of diabetes, the pre treatment duration of diabetes, the number of retinopathy lesions and the number of laser applications given to treat the maculopathy. RESULTS: On average visual acuity was unchanged at the post treatment control (mean=-0.04, SD=0.15), however, with a wide variation (range: -0.44 to 0.33). There was no correlation between the change in visual acuity and any of the studied background parameters. CONCLUSIONS: It is concluded that the treatment intervention rather than the general disease state is the main determining factor for the visual prognosis after laser photocoagulation for diabetic maculopathy. Other parameters should be identified to act as a basis for differentiating and improving laser photocoagulation of diabetic maculopathy. One such possible parameter might be the distance of retinopathy lesions and laser applications from the retinal fixation area. PMID- 11037912 TI - Grafting of the posterior cornea. Description of a new technique with 12-month clinical results. AB - PURPOSE: To describe the technique of grafting only the posterior cornea and to report 12-month clinical results. METHOD: A two-layer technique with an anterior recipient flap created by a microkeratome and a posterior penetrating donor graft allows for a watertight wound closure and at the same time a peroperative correction of astigmatism. Four eyes (3 patients) were followed for 12 months. RESULTS: The surgical technique could be completed in all cases without complications. The postoperative course was uneventful. The intrastromal absorbable sutures disappeared spontaneously and completely. Graft thickness showed the expected 6-month minimum while recipient flap thickness remained constant. After 1 year endothelial cell densities were 1200-2300 cells/mm2. Confocal microscopy showed activated keratocytes in the flap and quiescent keratocytes in the donor tissue by one year. The anterior chamber depth was normal in all cases. The optical quality of the cornea was studied by automatic keratometry and keratoscopy (TMS). The obtained optical properties were not optimal. CONCLUSIONS: The developed novel technique gives a better wound closure and a complication free postoperative course. It may allow for better control of postoperative astigmatism. In order to disseminate the use of the technique, eyebanks should supply posterior corneas to the surgeon. PMID- 11037913 TI - Orbital space-occupying lesions in Denmark 1974-1997. AB - PURPOSE: To determine the frequency, distribution according to diagnosis and the recurrence frequency of orbital space-occupying lesions in Denmark, to determine sex and age of the patients and to establish a national orbital data register. METHODS: All biopsied/surgically removed orbital lesions collected by Danish pathological departments during the period 1974-1997 were identified by SNOMED codes. In addition, in each case gender and age of the patients and number of recurrences were registered. RESULTS: 965 orbital lesions from 841 patients were identified during the 24-year period. The incidence of orbital lesions increased significantly over the study period and at present about 80 cases/year are registered. The ratio benign/malignant lesion per year remained constant during the observation period. Lesions in children constituted a total of 152 (16%), the percentage of lesions in children being constant during the period. Malignant tumors comprised a total of 364 (45%) in adults and 34 (22%) in children. Malignant tumors were in adults distributed with 43% primary, 48% secondary invading and 9% metastatic tumors, and in children with 65% primary, 29% secondary invading and 6% metastatic tumors. Normal tissue was only found in 6% of the cases recorded. CONCLUSION: A registration of all histologically evaluated orbital space-occupying lesions in Denmark during a period of 24 years was performed and a national orbital database established. Orbital lesions are rare and primarily benign. PMID- 11037914 TI - Visually impaired Swedish children. The 1980 cohort study--a 19-year ophthalmological follow-up. AB - PURPOSE: To assess the ophthalmological outcome in a cohort of visually impaired children in a geographically defined area. METHODS: A 19-year follow-up of medical records, interview and, if needed, re-examination of the initial 128 patients, born 1962-1976. RESULTS AND CONCLUSION: Follow-up was possible in 123 patients (96%); 76 (59%) were still visually impaired, 17 (13%) were deceased, while 30 (23%) had acquired a visual function > or = 0.3. The chances of gaining a visual function outside the WHO limits for visual impairment was significantly higher for patients without additional impairments (p=0.0023). The initial ophthalmologic diagnosis remained unchanged in 88%. The diagnoses with improved visual development included albinism and congenital nystagmus, while retinal diseases showed poorer results. An increase in visual function could be seen even in the initially older age-groups, indicating a maturation of visual function beyond what is usually considered the limit of plasticity of the visual system. PMID- 11037915 TI - Visually impaired Swedish children. The 1980 cohort study--aspects on mortality. AB - PURPOSE: Assessment of the burden of mortality in a cohort of visually impaired children in a geographically defined area. METHODS: A 19-year follow-up of medical records and death certificates. RESULTS AND CONCLUSIONS: Of the initial 128 patients, born 1962-76, 17 (13%) had died, giving a highly significant increase in mortality compared to the equivalent age-specific Swedish population (p-value <0.000001). All deceased had additional impairments. Respiratory causes of death were found in 12/17 cases. The primary ophthalmological diagnosis was cerebral visual impairment in 8 cases, optic atrophy in 6 cases, one each of congenital cataract, microphthalmus and chorioretinitis. A more pronounced level of visual impairment gave an increased risk of mortality. A significantly higher mortality rate among multiply impaired was seen in the combined group with 'childhood blindness' (visual acuity <0.05) and 'visual impairment, undetermined or unknown' opposed to the group with 'low vision' (visual acuity 0.05-<0.3) (p=0.047). PMID- 11037916 TI - Refractive results of radial keratotomy: a ten-year retrospective study. AB - PURPOSE: To investigate the long-term effects and stability of refraction after radial keratotomy procedure. METHODS: Radial keratotomy was performed on 123 persons to reduce myopia (range: -1 to -13 diopters) in 1986 to 1989. A mean of 11.5 years later (range 10 to 13), 61 of these patients with 102 eyes underwent a standardised refractive examination where subjective spherical equivalent refraction was measured and compared to the preoperative and the one month postoperative refractive measurement collected from the patients records. RESULTS: There was a reduction in spherical equivalent from an average of -5.46 diopters (SD 2.38) preoperatively to -2.32 diopters (SD 1.96) 11.5 years postoperatively. The mean change in direction of myopia between 1 month and 11.5 years postoperatively was 0.17 diopters (SD 1.18). This change was not statistically significant. From 1 month to 11.5 years, 10 of the eyes had developed more than 1 diopter hyperopia, and 20% more than 1 diopter myopia. When asked directly, all patients were satisfied with the result of their operation in general; 2 patients still complained of glare. CONCLUSION: No significant changes in refraction were found between 1 month and 11.5 years after radial keratotomy. Previously reported long-term studies on this field have found a trend toward progressive hyperopia. No evidence of such change can be supported by this study. PMID- 11037917 TI - Ophthalmological findings in children with congenital toxoplasmosis. Report from a Swedish prospective screening study of congenital toxoplasmosis with two years of follow-up. AB - Congenital toxoplasmosis may lead to severe visual impairment or neurological sequelae in the child. PURPOSE: To study the severity of the primary and late ophthalmological dysfunction during a prospective incidence study of congenital toxoplasmosis in the Stockholm and Skane counties. METHODS: Blood collected on phenylketonuria (PKU) cards from 40,978 consecutively born children were investigated for antitoxoplasma antibodies. Children with verified congenital toxoplasmosis were treated for 12 months with antiparasitic therapy and followed ophthalmologically, neurologically and serologically every third month. RESULTS: Three children had congenital toxoplasmosis. Two of these were asymptomatic at birth and would have escaped early detection without screening. One child had unilateral severe visual impairment and CNS involvement. The incidence of congenital toxoplasmosis was less than 1:10,000. CONCLUSION: Neonatal screening is of importance to diagnose asymptomatic infected children with congenital toxoplasmosis as treatment has been shown to reduce long-term sequelae. Ophthalmological investigations should start early and continue in co-operation with paediatricians. PMID- 11037918 TI - Secondary glaucoma in patients with juvenile rheumatoid arthritis-associated iridocyclitis. AB - PURPOSE: The prevalence and management of glaucoma were evaluated in patients with juvenile rheumatoid arthritis (JRA)-associated iridocyclitis. METHODS: The records of 69 patients with JRA-associated iridocyclitis were reviewed. RESULTS: Twenty-nine (42%) of these patients had secondary glaucoma or ocular hypertension. Glaucoma was controlled with topical treatment in only 7 of the 41 affected eyes (17%); systemic carbonic anhydrase inhibitor therapy resulted in control of another 8 eyes. Surgery controlled all but one of the remainder. CONCLUSION: Glaucoma is a common complication of JRA-associated iridocyclitis. It results from prolonged, inadequately treated intraocular inflammation and in some instances, from steroid use. Medical and surgical therapy for the glaucoma associated with JRA-uveitis is challenging and incompletely effective. We suspect that a more aggressive approach to the treatment of JRA-associated uveitis, earlier in the course of the disease may reduce this vision robbing contribution to the process. PMID- 11037919 TI - Visual field defects in patients taking vigabatrin. AB - PURPOSE: To investigate visual field loss in patients on long-term treatment with the antiepileptic drug vigabatrin, recently reported to cause visual disturbances. METHODS: Eighteen patients taking vigabatrin for 0.5-9.5 years were examined with automated perimetry up to 60 degrees from fixation using the Humphrey Field Analyser. Five patients with epilepsy receiving other medications served as controls. Patients found to have a visual field defect underwent ophthalmologic examination. RESULTS: Among the 18 patients in the vigabatrin group, visual field defects categorised as mild were revealed in 6 right eyes (33%) and 8 left eyes (44.4%), while defects categorised as severe were found in 9 right eyes (50%) and 8 left eyes (44.4%). The majority of the defects (66.7% in the right eye) were peripheral constriction with nasal predominance. The location of the defects was confirmed in 8 patients also tested with Kowa AP340 perimetry. CONCLUSION: According to our results, visual field defects among the patients on vigabatrin therapy may occur more frequently than previously recognised. PMID- 11037920 TI - Ocular changes in a case of Kyrle's disease. 20-year follow-up. AB - PURPOSE: To re-evaluate a case of Kyrle's disease with conjunctival and corneal changes in a 52-year-old patient with onset at the age of 5 years. METHODS: Repeated biopsies from the skin, conjunctiva and oral mucosa along with a corneal button obtained from a right eye keratoplasty were examined histopathologically. RESULTS: The skin, oral, conjunctival and corneal histopathology showed parakeratosis, dyskeratotic cells and single cell keratinization. The clinical picture and the histopathological findings are consistent with the diagnosis of Kyrle's disease. CONCLUSIONS: This is the first reported case of Kyrle's disease with conjunctival changes. We suggest that Kyrle's disease, although predominantly affecting young adults, may also be seen in childhood. The disease may be a genodermatosis. It is not confined only to the skin, but can be seen in mucous membranes like the conjunctiva and buccal mucosa, and in tissues having the same embryological origin as the skin, i.e. the cornea. PMID- 11037921 TI - Anterior uveitis following combined vaccination for measles, mumps and rubella (MMR): a report of two cases. AB - PURPOSE: To describe two children who developed anterior uveitis after vaccination for common childhood diseases. METHODS: A retrospective study of two Saudi patients who were seen at The Eye Center, Riyadh, Saudi Arabia. RESULTS: Aged 12 and 14, respectively, the two patients developed anterior non granulomatous uveitis 6 and 4 weeks after having the combined vaccination for measles, mumps and rubella (MMR). No other definite cause of uveitis was detected from history or from laboratory work up. Both patients responded to traditional regimens of uveitis therapy. However, they required a treatment period of several months. CONCLUSION: Anterior uveitis may occur following MMR vaccination. Cases of vaccination thus require ophthalmic awareness if visual symptoms or a painful red eye develop. PMID- 11037922 TI - Retinitis pigmentosa associated with peripheral sea fan neovascularization. AB - PURPOSE: To describe a case with retinitis pigmentosa associated with sea fan type retinal neovascularization. METHODS: Complete ocular examination including fluorescein angiography was performed in a 9-year-old girl. RESULTS: Ophthalmoscopically, in addition to arteriolar narrowing and bone corpuscular pigmentation of both retinae, a vascular lesion with surrounding intraretinal exudation was noted in the upper equatorial region of the right eye. On fluorescein angiography, the lesion stained in the form of a sea fan neovascularization. CONCLUSION: Sea fan type of neovascularization can be seen in association with retinitis pigmentosa. Fluorescein angiography is important in identifying the exact nature of such a lesion. PMID- 11037923 TI - Central retinal vein occlusion and HELLP syndrome. AB - PURPOSE: To present a rare case of central retinal vein occlusion in conjunction with the HELLP syndrome. METHODS: A 30-year-old woman presented in the 28th week of her second pregnancy with severe pre-eclampsia with HELLP syndrome; delivery by caesarean section was recommended. Ten days later, the patient complained of severely decreased visual acuity in her right eye. RESULTS: Ophthalmoscopy revealed a central retinal vein occlusion with venous engorgement and tortuosity, multiple flame hemorrhages, and disc and macular edema. Electroretinography revealed a reduction of b-wave/a-wave ratio. Fluorescein-angiography showed a blockage due to extensive retinal hemorrhages with late mild staining of the walls of veins. The patient presented a spontaneous improvement in visual acuity (0.8 two months after) and a complete resolution of ophthalmoscopic findings. CONCLUSION: Ophthalmic complications are possible during and soon after this syndrome. This is the first description of a patient suffering a central retinal vein occlusion during puerperium after the HELLP syndrome. PMID- 11037924 TI - Complicated eyelid reconstruction after an unusual glabellar flap repair. AB - Remote flaps may be used for lid reconstruction when tissue loss is extensive and there is insufficient tissue in the adjoining areas. Median forehead flaps are usually used for upper lid, medial canthal or nasal repairs. We describe a complicated reconstruction of the lid and correction of a deformity which resulted from the injudicious use of a glabellar flap for lower lid repair. Improper use of a median forehead flap may interfere with the functioning of the lid, leading to corneal exposure and poor cosmesis. Lower lid defects are better repaired by advancement flaps or techniques like Tenzel's semicircular flap, reverse Cutler Beard, Hughes procedure or Mustarde's repair. PMID- 11037925 TI - Extramedullary plasmacytoma of the orbit. AB - PURPOSE: To report a case of extramedullary plasmacytoma of the orbit. METHODS: A 71-year-old patient presented with diplopia eyelid fullness and limitation of ocular motility in the left eye. Visual acuity was counting fingers, intraocular pressure 34 mmHg and fundus eye examination showed choroidal folds in the involved eye. RESULTS: CT scan showed a mass filling the superior and external left orbit without bone destruction. A biopsy was performed revealing that the tumour was composed of plasmacytoid cells positive with immunohistochemical stains for Kappa light chains and epithelial membrane antigen. Systemic work up was negative. The diagnosis of extramedullary orbital plasmacytoma was made. The patient was treated with external beam radiotherapy (40 Gy) and has remained disease free for four years (49 months). CONCLUSION: Extramedullary plasmacytomas of the orbit are extremely rare tumours. Accurate and early diagnosis is essential for the therapeutic approach. Extensive medical work up to rule out multiple myeloma or other malignant lymphoproliferative conditions involving the orbit is needed when the diagnosis of orbital extramedullary plasmacytoma is suspected because treatment and prognosis are very different. PMID- 11037926 TI - Substituting latanoprost (Xalatan) for isopropyl unoprostone (Rescula) in monotherapy and combination therapy. PMID- 11037927 TI - Diabetic maculopathy. PMID- 11037928 TI - Corticosteroid treatment of facial haemangioma associated with sternal aplasia and supra-umbilical raphe. PMID- 11037929 TI - Endogenous panuveitis in a patient with Rickettsia conorii infection. PMID- 11037930 TI - Diversity and the state of sociological aging theory. AB - In the literature on aging there have been almost simultaneous calls for researchers to make more explicit links between theory and research and for researchers to incorporate diversity in their work. Although gerontologists have begun to document diversity, theory is often absent from this research. In this article, the author examines sociological aging theories of inequality and argues that this absence of theory may not be due to an oversight on the part of researchers. Rather, aging theories need to be rethought to be better suited for diversity research. PMID- 11037931 TI - Cognitive status and relocation stress: a test of the vulnerability hypothesis. AB - PURPOSE: This study investigated whether cognitively impaired nursing home residents are at particular risk of experiencing harmful effects during a mass, intra-institutional, interbuilding relocation. DESIGN AND METHODS: A pretest-post test experimental-comparison group design was used. Data on cognitive status, functional capacity, psychosocial health status, physical health status, and mortality were abstracted from the Minimum Data Set Plus and were analyzed using continuous and discrete survival analyses, controlling for covariates as well as baseline status of outcome variables. RESULTS: None of the Relocation x Cognitive Status interaction effects were significant. Relocation main effects indicated that movers in general were more likely than nonmovers to decline in physical health status. Evidence also emerged for a positive long-term effect of moving on psychosocial health status. IMPLICATIONS: These findings suggest cognitively impaired residents are not at unusual risk of harmful effects as a consequence of mass, interbuilding transfer. Given the significant relocation main effects, though, caution must be taken in moving cognitively impaired residents, as it should be in moving any residents. PMID- 11037932 TI - Memory complaints, coping, and well-being in old age: a systemic approach. AB - A study on memory complaints (as measured by selected subscales of the Metamemory in Adulthood Questionnaire) and its context was conducted on 179 older adults. A path analysis showed that memory complaints influence coping behavior through memory-related anxiety and perceived seriousness of complaints and that both memory complaints and coping influence well-being. Locus of control was found to be the most important antecedent variable in the model. PMID- 11037933 TI - Environmental press and adaptation to disability in hospitalized live-alone older adults. AB - PURPOSE: This study examined the ability of personal competency variables at the time of hospital discharge to predict primary instrumental activities of daily living (IADLs) and secondary outcomes (living arrangements) in a sample of 194 urban, live-alone, older adults who had a new onset disability. DESIGN AND METHODS: Consecutively admitted medical rehabilitation patients, 72% women and 85% African American, participated in the study. Using path analysis, three of the four competency variables collected at the time of hospitalization (cognition, medical burden, activities of daily living) predicted IADLs at 3 and 6 months after hospitalization (e.g., cooking, telephone use, money management). IADLs, in turn, predicted living arrangements at 3 and at 6 months after hospitalization. RESULTS: The findings provided strong support for the importance of assessing a broad range of competency variables when investigating adaptation to disability. IMPLICATIONS: The increased understanding of adaptation in live alone older adults with a new-onset disability is particularly timely given the increase in live-alone older adults and the dire consequences associated with change in living arrangement (i.e., mortality and morbidity) in this group. PMID- 11037934 TI - Hospital provision of institutional long-term care: pattern and correlates. AB - PURPOSE: This study examined the pattern and correlates of institutional long term care provision among U.S. community hospitals, differentiating two categories of services: (1) skilled nursing and rehabilitation (SN-R) and (2) other long-term care (O-LTC). DESIGN AND METHODS: Multinomial logistic regression analysis was used to examine the associations of hospital and community characteristics with the pattern of long-term care provision (SN-R only, O-LTC only, both SN-R and O-LTC, and None) among 3,842 hospitals. RESULTS: The pattern of long-term care provision was significantly associated with hospitals' mission (for-profit and teaching status) and their internal and external resources. IMPLICATIONS: Results suggest the importance of considering hospital and community characteristics in predicting the impact of policy changes and in envisioning the role of hospitals in long-term care. PMID- 11037935 TI - Does type of disease matter? Gender differences among Alzheimer's and Parkinson's disease spouse caregivers. AB - PURPOSE OF STUDY: Mental health outcomes are widely reported among spouse caregivers, with wives generally faring worse than husbands. We hypothesized that gender differences would not be as strong in a cognitively intact group because caring for cognitively intact spouses may involve less severe reciprocity losses. We also examined gender differences in coping strategies within each group. DESIGN AND METHOD: 175 spouse caregivers for patients with Alzheimer's disease (AD; n = 88) and Parkinson's disease (PD; n = 87) were interviewed. Participants completed perceived stress (PSS), depression (CES-D), state anxiety (STAI, Form Y), and coping strategies (WCCL-R) measures. RESULTS: Wives in the AD group reported significantly worse mental health outcomes than husbands, while wives and husbands in the PD group showed no differences. AD caregiving wives were less likely than husbands to use problem-focused coping strategies. There were no significant gender differences in either group for social support or emotion focused coping. IMPLICATIONS: Loss of reciprocity in marital relationships may affect women more negatively than men. Future studies that address underlying mechanisms of gender differences and focus on similar caregiving situations and contexts deserve attention. PMID- 11037936 TI - Assessment and understanding of pain in patients with dementia. AB - The literature on pain in dementia patients is reviewed. A summary of methods for assessment of pain in demented elderly persons and an examination of studies that used such methods are included. In addition, literature theorizing a decrease in affective pain in this population is discussed; management of pain in such patients is not discussed extensively. Research reveals 3 major findings: (a) a moderate decrease in pain occurs in cognitively impaired elderly persons, (b) communicative dementia patients' reports of pain tend to be as valid as those of cognitively intact patients, and (c) assessment scales developed thus far for noncommunicative patients require improvement in accuracy and facility. Many questions about pain in dementia patients remain, and the continued development of valid pain assessment techniques is a necessity. PMID- 11037937 TI - Stability of performance of activities of daily living using the MDS. AB - The Minimum Data Set (MDS) requires assessment of performance of activities of daily living (ADLs) by newly admitted nursing home residents over all shifts for a 7-day period, for a total of 21 assessments. This study evaluated within subject equivalence of multiple assessments of 42 residents' admission MDS ADL performance. Friedman two-way analysis of variance for ranks documented no significant within-subject differences among repeated measurements for all 13 MDS ADL variables. Thus, fewer than 21 assessments may accurately assess ADL performance. PMID- 11037938 TI - Therapy use and discharge outcomes for elderly nursing home residents. AB - This study examines therapy use and discharge outcomes (community discharge, mortality, or remaining in the facility) over a 90-day period for 1,419 elderly, post-acute care nursing home admissions in South Dakota. Subjects met criteria as rehabilitation candidates (i.e., absence of serious behavioral or medical conditions that would limit rehabilitation potential). Receipt of therapies was related significantly to age (younger), Medicare coverage, hip fracture or stroke diagnosis, absence of cancer diagnosis, and resident or staff expectations for functional improvement. Therapy use was related positively to community discharge and negatively to mortality when controlling for covariates such as age, marital status, payment source, functional status, cognitive status, and major diagnoses. Also, community discharge was related positively to the facility's volume of therapy provision and percentage of Medicare-covered stays. PMID- 11037939 TI - Private long-term care insurance and the asset protection motive. AB - This research examined the role of assets in the decision to purchase insurance for long-term care using survey data from the Asset and Health Dynamics Among the Oldest Old (AHEAD) study. Previous research suggests that assets matter, but the size and direction of the effect varies. An important issue regarding the role of assets has not been explored adequately--whether the effect of assets differs between less wealthy and very wealthy individuals. A methodology to control for this type of variation is employed in this analysis. Results suggest that increases in assets have the greatest influence on the probability that less wealthy individuals own long-term care insurance, and have a negligible impact on the wealthy. This has important implications for policies designed to increase long-term care insurance ownership. PMID- 11037940 TI - The balance of benefit: a review of intergenerational transfers in Australia. AB - PURPOSE: This article reviews the financial and nonfinancial transfers taking place intergenerationally and between older people and the community. DESIGN AND METHOD: Secondary data were used in the analysis and discussion to provide an overview of the Australian context. RESULTS: Within the public arena, governments provide major financial contributions through money transfers and the provision of residential support. Older people provide considerable community support by undertaking voluntary services. This article concludes that the balance of benefit is difficult to determine; however, in terms of public expenditure older people are major recipients. Within the family, the balance of benefit is reversed. Older people are major monetary contributors to adult children and their families in the transition to an independent status. Older people are also the principal carers of their frail-aged partners, thus reducing both the burden of care on their adult children and government institutions. IMPLICATIONS: The analysis reported here has major implications for the development of policy and structural change and for reducing negative stereotypes of dependency in old age. PMID- 11037941 TI - "Outings to your taste": a nutrition program for the elderly. AB - "Outings to Your Taste" is an innovative program that aims to improve the nutritional status and social network of elderly people who receive home delivered meals. This article examines participation in one of the program's components, outings to community restaurants. Participation data were collected on-site and information about client characteristics was collected in at-home interview surveys of targeted clients (n = 144). While about half of the clients had tried at least one outing, more than 25% of them participated in at least one third of the outings offered to them. Results indicate that the program attracted a variety of clients in terms of sociodemographic, health, and social isolation characteristics. PMID- 11037942 TI - The achievements of a multiservice project for older homeless people. AB - This report of the achievements of an experimental multiservice center in London for older street people begins with reviews of the types of long-term accommodation available for resettlement and the work of its outreach team, 24 hour open access rooms, and residential, assessment, and resettlement services. Two outcomes are examined: whether users returned to the streets and whether they were resettled in long-term housing. Those with alcohol dependency were most difficult to resettle. Logistic regression analyses of the factors influencing the two outcomes indicate that the duration of residence in the center was the the predominant influence. PMID- 11037943 TI - A novel gene: sawD related to the differentiation of streptomyces ansochromogenes. AB - A 1.3 kb DNA fragment was cloned from a total DNA library of Streptomyces ansochromogenes using Southern hybridization. Nucleotide sequencing analysis indicated that the 1320 bp DNA fragment contained a complete open reading frame (ORF). In search of databases, the deduced product of ORF containing 213 amino acids is homologous to the serine protease of Caulobacter cresceatus, and a conserved serine-catalytic active site (GPSAG) exists. The gene was designated as sawD. The function of this gene was studied with the strategy of gene disruption, and the result showed that the sawD may be related to sporulation and especially to the spore septation in Streptomyces ansochromogenes. The preliminary result indicated that sawD mutant could produce abundant pigment in contrast with the wild type, it seems that sawD gene may be involved in pigment biosynthesis, and this gene is also dispensable for biosynthesis of nikkomycin in Streptomyces ansochromogenes. PMID- 11037944 TI - Construction of plant vectors with high level expression of B.t. toxin gene and studies on their expression behavior in transgenic tobaccos. AB - Breeding pest-resistant plants using plant genetic engineering technique is an effective strategy in integrated pest management (IPM). Increasing the expression level of foreign insecticidal protein by using a strong promoter is a useful method. In this work, a plant expression vector, pBinMoBc, was constructed. It contained the CryIA(c) gene under the control of a chimeric OM promoter and the omega factor. As a control, another vector, pBinoBc, was also constructed in this study. pBinoBc carrying CryIA(c) gene was under the control of a CaMV 35S promoter. The vectors were transferred into tobacco plants via Agrobacterium mediated transformation. ELISA assay showed that the average expression level of the CryIA(c) gene in pBinMoBc transgenic tobacco plants is 2.44-times that in pBinoBc transgenic tobacco plants. The expression of B.t. toxin in individual plant can be up to 0.255% of total soluble proteins. Bioassay showed that pBinMoBc transgenic tobacco plants had more notable insecticidal effects than pBinoBc transgenic tobacco plants. The above results showed that the chimeric OM promoter is a stronger promoter than the CaMV 35S promoter that was widely used in plant genetic engineering. This is very useful in pest-resistant plant genetic engineering. PMID- 11037945 TI - Expression of transthyretin gene in Pichia pastoris. AB - Plasmid pSK-TTR was digested by BamHI and the DNA fragment containing Transthyretin gene was cloned into the BamHI site of vector pHIL-SI. The recombinant plasmid pHIL-SI-TTR was digested by BglII; the larger fragment was transformed into yeast Pichia pastoris. SDS-PAGE analysis showed that the yeast transformant can express and secrete TTR as a fusion protein, TTRF. To purify TTRF,the DEAE-Sepherose F.F. chromatography and Sephacryl S-200 chromatography were used. The in vitro test showed that TTRF could inhibit the growth of human hepatoma cells. PMID- 11037946 TI - Expression of rainbow trout growth hormone cDNA in Saccharomyces cerevisiae. AB - Rainbow trout growth hormone cDNA was modified by polymerase chain reaction (PCR). The modified cDNA was subcloned into the E. coli-yeast shuttle vector pMA91 under the yeast PGK promoter, and transformed into Saccharomyces cerevisiae Y33 to construct an expression strain Y33 (pMArGH16). The recombinant gene could express the growth hormone peptide (about 3% of the total yeast proteins) in Y33 (pMArGH16). The expression product was used as a supplement to feed Tilapia fingerlings. The result showed that the recombinant fish GH could significantly enhance the growth of Tilapia fingerlings. PMID- 11037947 TI - Gene fusion and expression of heat-labile and heat-stable enterotoxins of enterotoxigenic Escherichia coli. AB - The vaccine candidate comprising the genes that code the B subunit of the heat labile enterotoxin (LT-B) and the heat-stable enterotoxin (ST) of enterotoxigenic E. coli (ETEC) had been constructed by recombinant genetic techniques. The 5'terminus of the gene encoding pro-ST was genetically fused to the 3'terminus of the LT-B gene. The pro-ST gene containing mature ST sequence and pro sequence which codes for the pro region of ST precursors was amplified by PCR from pSLN004 plasmid. To reduce toxicity of the ST in vitro was substituted Leu for Ala residue at position 14 of ST by oligonucleotide-directed site mutagenesis. For this construction, the expression of ST antigenicity and LT antigenicity were obtained when a five amino-acid or a nine-amino-acid linker were included between the LT-B and pro-ST moieties. The LT-B/pro-ST fusion peptides possessed no enterotoxic activity of heat-stable and heat-labile enterotoxins, and retained the ability to bind GM1 ganglioside. More importantly, these LT-B/pro-ST fusion peptides were immunogenic. The preparations containing the hybrid molecule elicited special antibodies that were to recognize native toxin in vitro. PMID- 11037948 TI - Optimization of conditions for safflower cell culture and accumulation of cellicolous product tocopherols. AB - The culture of calli, which were induced from cotyledon of Carthamus tinctorius L, was able to synthesize tocopherols. Results determined by NP-HPLC showed that the main component was alpha-tocopherol. When 0.1% casein was added into MS medium, and 2,4-D concentration was adjucted to 0.30 mg/L and 6-BA to 1.80 mg/L, alpha-, beta- and gamma-forms of tocopherol reached 4.1706, 0.3120 and 0.1650 microg/g fresh calli, respectively. The effective tocopherol content was 4.4091 microg/g fresh calli, and was 57 times higher than that in the control. PMID- 11037949 TI - Study on rt-PA production by recombinant CHO cells immobilized with a porous microcarrier. AB - Using the porous microcarrier Cytopore, immobilized cultivation of a recombinant CHO cell line 4B3 producing tissue type plasminogen activator (rt-PA) was performed in a 3L stirring tank bioreactor with the perfusion of serum-free medium DF5S. 4B3 cells could enter the inner space and grow both in the inner space and on the surface of the Cytopore. The highest viable cell density of 4B3 and the highest production level of rt-PA were 8.83 x 10(6) cells/mL and 12,473 IU/mL, respectively. Purified by MPG adsorption chromatography and lysine Sepharose 4B affinity chromatography, the purified rt-PA with a purity exceeding 98% was obtained from the culture supernatant. PMID- 11037950 TI - Continuous ethanol fermentation using the fusion strain SPSC (I) flocs: floc size distribution, cell growth, and ethanol fermentation kinetics. AB - Based on the description for the floc size distribution, the superficial kinetics of the SPSC(I) floc growth and ethanol production were obtained. mu' = 0.134S/19.23+S for the flock growth and v' = 0.747S/10.871+S [(1- p/126.3)(2.593)] for the floc ethanol production. PMID- 11037951 TI - Detection and nucleotide analysis of RNA5 of beet necrotic yellow vein virus isolated in China. AB - Using the RT-PCR method, five isolates of beet necrotic yellow vein virus from China were used for the RNA5 genomic component detection and sequence analysis. The result showed that RNA5 was only found in the isolates from Baotou and Hohhot, but not in those of Xingjiang, Helongjiang, and Wulate of Inner Mongolia. The RNA5 components had 1338 nucleotides and 1358 nucleotides in the length of Baotou and Hohhot isolates, respectively, in which the single open reading frame (ORF) encoding for a proteins of 26kD were contained. Compared with the published sequences of F72 and D5 isolates, these RNA5 components shared the identity of 93.7% approximately 98.5% in nucleotide acids and 91.8%-98.2% in deduced amino acids. PMID- 11037952 TI - Management of pain and pain-related symptoms in hospitalized veterans with cancer. AB - Unrelieved pain continues to be a problem among hospitalized patients with cancer. The purpose of this study was to evaluate pain management outcomes in a group of veterans with cancer receiving inpatient care. The sample consisted of 90 veterans with cancer hospitalized in one of two large veterans medical centers in the southeastern United States. Daily pain was assessed by administering the visual analog scale (VAS) for pain three times in a 24-hour period and averaging these three scores. The Brief Pain Inventory (BPI) and Constipation Assessment Scale (CAS) were administered once. The charts were audited using the Chart Audit for Pain (CAP). The sample was predominantly male (93.3%) and white (82.8%). The length of time since diagnosis ranged from newly diagnosed during this hospitalization to 16 years. Average daily pain was 32.9 on the VAS and 4 on the BPI. However, approximately one-fourth of the patients reported average daily pain above the midpoint (VAS > 50), and some patients reported average daily pain to be as high as 98. Fewer than half of charts (42%) showed evidence that a pain rating scale was used. Other assessment data also were very limited. Patients reported that pain interfered with all activities on the BPI, with highest interference scores for walking and sleep (mean, 5.5). Although 80% of the patients reported some problem with constipation, the chart audit indicated that this was recorded in only 11 patient records. No patient records indicated a problem with sedation. The findings indicate that limited attempts were made to manage pain using nonpharmacologic methods. In addition, only one of the nine charts reporting these attempts showed evidence that results from the attempt were evaluated. It may be concluded that pain management continues to be less than ideal in these veterans hospitals. Study results indicate that nurses are not documenting careful assessment of pain, not documenting evaluation of approaches to pain management, and not attending to the constipation that is inevitable when opioids are administered. Continued emphasis on nursing education related to pain management is needed. Future research should be undertaken to evaluate these outcomes. PMID- 11037953 TI - A clinical profile to predict decision making, risk behaviors, clinical status, and health-related quality of life for cancer-surviving adolescents: part 2. AB - This is the second part of a two-part article describing a multifactorial model of clinical factors predicting decision-making quality, risk behaviors, clinical status, and health-related quality of life for cancer-surviving adolescents. To support the conceptualization of the model, findings from the literature and from the research program of the current author are presented. In part 1, support for the antecedent predictors, both primary and secondary factors, was presented. In part 2, the mediator of decision making, the moderator of risk motivation, and the expected outcomes related to risk behaviors, clinical status, and health related quality of life are addressed. Besides a description supporting the second part of the clinical profile and its empirical underpinnings in part 2, methodologic challenges in future research and implications for clinical trials and clinical use specific to cancer-surviving adolescents also are discussed. PMID- 11037955 TI - Sociodemographic predictors of adherence to annual cervical cancer screening in minority women. AB - The Papanicolaou (Pap) test is an effective screening mechanism for reducing morbidity and mortality from cervical cancer. Nevertheless, Pennsylvania ranks fifth in national cervical cancer incidence and fourth in national cervical cancer mortality, with a significant number of cases contributed by Philadelphia. Substantial subgroups of American women, specifically ethnic minorities, the elderly, the uninsured, and the poor, have not been screened or are not screened at regular intervals. A secondary data analysis was conducted to test whether age, income, insurance coverage, marital status, level of education, and number of persons living at home could predict whether a woman among convenience sample of 204 black and Hispanic women adhered to annual Pap testing. A woman was considered adherent to annual Pap testing if she reported undergoing a Pap smear in the 14 months preceding her enrollment in the study. African American woman who were high school graduates and had insurance coverage were more likely to be adherent to annual Pap testing. Hispanic women older than 50 years and born outside the mainland United States were less likely to be adherent to annual Pap testing. Findings suggest that cancer nurses working to promote cervical cancer screening in Philadelphia should continue to target at-risk populations, specifically uninsured and less-educated black and Hispanic women older than 50 years who were born outside the mainland United States. PMID- 11037954 TI - Anger and cancer: an analysis of the linkages. AB - The purposes of this article are to review literature on anger's link to cancer, to analyze the state of the science in this area, and to propose some directions for future research. Extremely low anger scores have been noted in numerous studies of patients with cancer. Such low scores suggest suppression, repression, or restraint of anger. There is evidence to show that suppressed anger can be a precursor to the development of cancer, and also a factor in its progression after diagnosis. Some studies indicate that it may be beneficial for patients to mobilize anger to battle their cancer. However, there is a paucity of research on the outcomes of various anger interventions. Longitudinal studies that repeatedly measure anger and other moods over the disease trajectory are needed. PMID- 11037956 TI - Women's perspectives regarding the impact of ovarian cancer: implications for nursing. AB - Ovarian cancer is the fourth leading cause of cancer-related death in women. Ovarian cancer and its treatment have a considerable effect on the quality of life of women diagnosed with the disease. Currently, little is known about the perspectives of women regarding their experiences of living with ovarian cancer or the impact of recurrent disease. This article presents data from a national study of Canadian women living with ovarian cancer and describes the impact of the disease and its treatment. In this study, 93 women had recurrent disease, and 170 had not experienced recurrent disease. Women in both groups were similar, ranging in age from 21 to 61 years. Two-thirds of the women were married, and all were white. A greater proportion of the women with recurrent disease reported bowel problems; fears of dying, pain, getting around; and feelings of self-blame. On the average, women with recurrent disease reported experiencing more problems since diagnosis than those without recurrent disease (p = 0.01). The proportion of women who perceived that they received adequate help for their problems ranged from 20% to 85%. Implications for oncology nurses regarding assessment, referral for assistance, and patient education are apparent from the study findings. PMID- 11037957 TI - Meanings of dying at home for Chinese patients in Taiwan with terminal cancer: a literature review. AB - To maintain dignity, patients with terminal cancer must be able to do things in their own way, to make their own decisions, and to preside over their own dying. Among the tasks considered essential for patients with terminal cancer is deciding where they prefer to die. The actual place of death has been recognized in hospice care as indicating quality of care. Approximately two-thirds of patients with cancer, when asked about the preferred place of death, say they wish to die in their own homes. Patients with terminal cancer dying at home may find physical and emotional comfort there. Home is a place where people may feel safety and a sense of belonging. In dying at home, patients with terminal cancer also may have a greater chance to control their environment, more autonomy and privacy, and a sense of normality. In this article, special cultural meaning of dying at home for the Chinese patient and the family is reviewed. It is essential for health care professionals to understand Chinese cultural beliefs and values related to dying at home in order to provide culturally sensitive care for Chinese dying patients and their families and to enhance their sense of control over the unknown process of dying. PMID- 11037958 TI - A comparison of nurse and patient perceptions of chemotherapy treatment stressors. AB - To assess how closely nurses and patients considered various situations as causing stress in chemotherapy treatment, this study was designed to compare nurse and patient perceptions of chemotherapy treatment stressors for patients in an oncology hospital. This descriptive study was conducted in the summer of 1997. Fifty patients receiving cytotoxic treatment were chosen by random sample. All 21 registered nurses of the Tabriz University of Medical Sciences who worked in the oncology department were contacted. A Likert-type questionnaire was designed to measure the stressfulness of commonly occurring items in chemotherapy treatment. The intensity of each item was measured by a scale with response choices ranging from the most to the least important. The patients were asked to indicate their perceptions of the seriousness of stress by sorting items. The nurses were asked to complete the same questionnaire as the patients completed. The greatest physical stressor mentioned by the patients was fatigue (66%), and by nurses, alopecia (62%). The nurses' perceptions of psychosocial items causing the greatest stress included fear of disease recurrence (90.5%), fear of death (90.5%), economic problems (90.5%), and appearance changes (90.5%), whereas the patients perceived dependency (80%), economic problems (70%), and loss of social activity (66%), as the greatest stressors (p < 0.05). The findings showed disagreement between the two groups on intensity of physical and psychosocial stressors (p < 0.05). The results from this study increased nurses' awareness of important stress factors in chemotherapy treatment. Identification of the situations perceived to be more stressful than others helped clinical nurses to modify their care and provide for their patients in a way that removes or reduces the stressors. PMID- 11037959 TI - Cancer nursing research priorities: a Norwegian perspective. AB - The purpose of the study was to determine research priorities among Norwegian nurses in cancer care, and to investigate implications that these priorities might have for future planning of nursing research. Differences between specialists in cancer nursing and other nurses working in cancer care, and between the current results and earlier findings in this area also were evaluated. Half the members of The Norwegian Society of Nurses in Cancer Care (n = 197) were mailed a questionnaire used in a similar Canadian study. The nurses were asked to select the five topics they perceived as most important from a list of 80 items, and to rank them in order of research priority. The response rate was 43% (197/464), and 75 respondents were specialists in cancer nursing. Quality of life was given the highest research priority in the total sample. Psychosocial support/counseling, communication between patient and nurse, patient participation in decision making, nurse burnout, and ethics also were ranked highly. In contrast to the others, cancer nursing specialists ranked ethics as their number one priority. Except for symptom management, the priorities given in Norway and other Western countries were found to be similar. These results might suggest topics for future research tailored to the needs of cancer nursing. PMID- 11037960 TI - The significance of fellow patients for the patient with cancer: what can nurses do? AB - Based on two qualitative studies, this article describes the significance of fellow patients in cancer wards, and the relevance for health personnel involved in patient-patient relationships is discussed. Using grounded theory, in-depth interviews were conducted in the first study with 21 adult cancer patients and in a follow-up study with 8 patients and 27 health care providers. The findings indicate that the significance of fellow patients for the patient with cancer can be described in terms of three different dimensions: attitudes toward own illness, interpersonal relationships, and environmental factors. Although contact with fellow patients most often seems to engender positive experiences, some negative experiences are reported also. The suggestions in this article for practical implications are meant to help nurses and other health care providers to promote positive and prevent negative consequences in patient-patient relationships in hospitals. PMID- 11037961 TI - Midwives' descriptions of their familiarity with cancer: a qualitative study of midwives working with population-based cervical cancer screening in urban Sweden. AB - Nurse-midwives are responsible for taking Papanicolaou (Pap) smears in Swedish population-based cervical cancer screening programs. A research project examining the screening program from the perspective of different stakeholders includes an interview study of 21 midwives working in Stockholm. This article explores the way the midwives describe cancer-related knowledge and aspects of screening, contrasting this with relevant findings from a substudy of 66 healthy women participating in screening. A semistructured interview guide with open-ended questions was used to investigate ideas about benefits and risks in the screening program, risk factors for cervical cancer, the reliability of the test itself, sources of information/knowledge relevant for cervical cancer screening, and the manner in which the midwife described her role in the screening program. The transcripts of the audiotaped interviews were analyzed thematically using a team approach. The interviewed midwives showed a great deal of consensus in their descriptions of lacking familiarity with cervical cancer and its prevention and treatment. The midwives said they lack recent education and knowledge, often avoiding use of the word "cancer" with women attending screening. It seems that the midwives experienced little professional guidance in discussing cancer related issues with women attending the screening program. In this study, they appeared to rely on personal knowledge, values, and experience instead. PMID- 11037962 TI - Changes in public health nurses' knowledge and perception of counseling and clinical skills for breast and cervical cancer control. AB - This study discusses the findings from two surveys of 459 nurses working in public health about their knowledge and perception of counseling and clinical skills related to breast and cervical cancer control. The first survey was conducted in 1993 before professional education efforts with the Breast and Cervical Cancer Control Program (BCCCP) began in North Carolina. The second survey was carried out in 1996. The results show a significant increase from 1993 to 1996 in recognition of increasing age, nulliparity, late childbearing, and family history as risk factors for breast cancer, and for smoking and early age at first sexual intercourse as risk factors for cervical cancer. Nurses' skills were more likely to be rated as excellent or very good in 1996 as compared with ratings in 1993 for educating women about mammography, performing a clinical breast examination, teaching breast self-examination by demonstration, performing a Papanicolaou (Pap) smear and bimanual examination, and counseling women about abnormal Pap tests. Nurses who took a BCCCP-sponsored adult physical assessment course were more likely to know correct risk factors for breast and cervical cancer, and to show improvement in teaching breast self-examination, performing a Pap smear, and counseling women about abnormal Pap tests. PMID- 11037963 TI - Leptin in the regulation of immunity, inflammation, and hematopoiesis. AB - Leptin, the product of the ob gene, is a pleiotropic molecule that regulates food intake as well as metabolic and endocrine functions. Leptin also plays a regulatory role in immunity, inflammation, and hematopoiesis. Alterations in immune and inflammatory responses are present in leptin- or leptin-receptor deficient animals, as well as during starvation and malnutrition, two conditions characterized by low levels of circulating leptin. Both leptin and its receptor share structural and functional similarities with the interleukin-6 family of cytokines. Leptin exerts proliferative and antiapoptotic activities in a variety of cell types, including T lymphocytes, leukemia cells, and hematopoietic progenitors. Leptin also affects cytokine production, the activation of monocytes/macrophages, wound healing, angiogenesis, and hematopoiesis. Moreover, leptin production is acutely increased during infection and inflammation. This review focuses on the role of leptin in the modulation of the innate immune response, inflammation, and hematopoiesis. PMID- 11037964 TI - The protective effect of IFN-gamma in experimental autoimmune diseases: a central role of mycobacterial adjuvant-induced myelopoiesis. AB - The study of animal models for organ-specific autoimmune disease contributes to our understanding of human diseases such as multiple sclerosis and rheumatoid arthritis. Although experimental autoimmune diseases develop spontaneously in certain strains of mice, others need to be induced by administration of organ specific autoantigen, often together with complete Freund's adjuvant (CFA), containing heat-killed mycobacteria. In the two types of models, the role of endogenous interferon-gamma (IFN-gamma) has extensively been investigated by using neutralizing anti-IFN-gamma antibodies and by employing mice genetically deficient in IFN-gamma or its receptor. In these studies disease-promoting as well as disease-protective roles of endogenous IFN-gamma have been described. Remarkably, in most models that rely on the use of CFA, there is abundant evidence for a protective role. Here, we review evidence that this role derives from an inhibitory effect of IFN-gamma on myelopoiesis elicited by the killed mycobacteria. These findings explain the bimodal role of IFN-gamma in different models of autoimmune disease and raise questions regarding the clinical relevance of these models. PMID- 11037965 TI - The immunobiology and clinical potential of immunostimulatory CpG oligodeoxynucleotides. AB - Over 100 years ago, Coley first explored the use of bacterial products as immunostimulatory therapy for nonbacterial disease. It is now clear that bacterial DNA, and synthetic oligodeoxynucleotides containing specific motifs centered on a CpG dinucleotide (CpG ODN), are potent immunostimulatory agents. The molecular mechanisms responsible for the immunostimulatory effects of CpG ODN have yet to be elucidated fully, although it is clear that CpG ODN act rapidly on a variety of cell types. This includes activation of B cells, natural killer cells, and antigen-presenting cells including monocytes, macrophages, and dendritic cells. These effects have led to evaluation of CpG ODN as immune adjuvants in immunization where they have been shown in animal models to enhance the development of a TH1-type immune response. Preliminary results from clinical trials using CpG ODN as an immune adjuvant are promising. Preclinical studies suggest CpG ODN can also enhance innate immunity against a variety of infections, synergize with monoclonal antibody to enhance antibody-dependent cellular cytotoxicity, and alter the Th1/Th2 balance as a possible treatment for allergic diseases and asthma. Clinical evaluation has recently begun to determine whether promising preclinical results with CpG ODN can be translated into effective and tolerable clinical treatment approaches. PMID- 11037966 TI - Impaired splenic erythropoiesis in phlebotomized mice injected with CL2MDP liposome: an experimental model for studying the role of stromal macrophages in erythropoiesis. AB - Erythropoiesis occurs in the presence of erythropoietin (EPO) without macrophages in vitro. In hematopoietic tissues, however, erythroid cells associate closely with stromal macrophages, forming erythroblastic islands via interactions with adhesion molecules. To elucidate the role of macrophages in erythropoiesis, we selectively abrogated stromal macrophages of splenic red pulp of phlebotomized mice by injection with dichloromethylene diphosphonate encapsulated in multilamellar liposomes (CL2MDP-liposome). In the spleen, no erythropoietic activity occurred until 5 days after the treatment. Colony assay revealed that the erythropoiesis was suppressed at the level of CFU-E. The splenic erythropoietic activity gradually developed from day 6 after the treatment, when F4/80+ macrophages began to appear in the red pulp. EPO mRNA was expressed in kidney but not in liver or spleen of phlebotomized mice injected with CL2MDP liposome, and the serum EPO concentration in these mice was higher than that in phlebotomized mice. These findings suggest that abrogation of stromal macrophages by injection with CL2MDP-liposome impairs the splenic microenvironment for erythropoiesis induced by hypoxic stress, and this may be an excellent experimental model for further characterization of the in vivo role of splenic macrophages in erythropoiesis. PMID- 11037967 TI - Migration of dermal cells expressing a macrophage C-type lectin during the sensitization phase of delayed-type hypersensitivity. AB - Dermal cells expressing a macrophage C-type lectin (mMGL) were previously suggested to migrate to regional lymph nodes during the sensitization phase of delayed-type hypersensitivity (DTH). The migration seemed to be induced by the solvent used to dissolve the antigen, and the DTH response was significantly enhanced by the migration. In this study, immunohistochemical analysis of skin after epicutaneous application of one of such solvents, a mixture of acetone and dibutylphthalate (AD), revealed a transient decrease in the number of mMGL positive cells in the dermis. A similar decrease in this cell population was also observed in an ex vivo assay with skin explants excised from AD-treated sites. Conditioned medium from organ culture of AD-treated skin induced a similar decrease of mMGL-positive cells in untreated dermis, indicating the involvement of soluble factors. mMGL-positive cells seemed to represent a unique subpopulation of F4/80-positive dermal cells. PMID- 11037968 TI - Functional consequences of FcepsilonRIalpha up-regulation by IgE in human basophils. AB - These studies examine the functional changes that occur after up-regulation of FcepsilonRIalpha by immunoglobulin E (IgE) for human basophils. Basophils were cultured with and without IgE antibody (PS myeloma IgE or anti-gp120-specific IgE) for 1 week and challenged with anti-IgE, anti-FcepsilonRIalpha, or antigen for histamine and IL-4 secretion. There were no statistically significant changes in their response to anti-IgE or anti-receptor antibodies, as compared with controls incubated for the same period, whereas receptor expression increased an average of 4-fold. There was increased responsiveness to antigenic challenge, most notably at suboptimal concentrations of antigen (gp120 peptide-ovalbumin conjugate). For a 6-fold difference in cell surface density of gp120-specific IgE, there was a 2.2-fold change in antigen potency or 3-fold increases in histamine release at lower antigen concentrations. Similar results were found for secretion of IL-4. Basophil sensitivity, which is a measure of the density of antigen-specific IgE required for 50% of maximal secretion, was used to determine whether up-regulation of FcepsilonRIalpha was coordinated with up-regulation of other components of the IgE-signaling pathway. The results indicated up regulation of FcepsilonRI is not always accompanied by changes that allow sensitivity to be maintained. These results indicate that functional up regulation does occur but that its magnitude may be modulated because not all components of the signaling pathway are up-regulated in a balanced manner. PMID- 11037969 TI - Altered membrane trafficking in activated bone marrow-derived macrophages. AB - Activation of macrophages with interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS) leads to increased intracellular resistance to microbes and increased major histocompatibility complex class II-restricted antigen presentation, processes that both use the vacuolar compartment. Despite the requirement of the macrophage vacuolar compartment for microbicidal activities and antigen processing, the rates of endocytosis and membrane trafficking in activated macrophages are not clearly defined. In this study, vacuolar compartment dynamics were analyzed in murine bone marrow-derived macrophages activated with LPS and/or IFN-gamma, conditions that increased macrophage nitric oxide production and resistance to infection by Listeria monocytogenes. Relative to nonactivated cells, activated macrophages showed diminished rates of fluid phase pinocytosis and phagocytosis and delayed progression of macropinosomes and phagosomes to late endosomes and lysosomes. In contrast to the slowing of membrane trafficking, rates of macropinosome acidification were similar between activated and nonactivated cells. One consequence of this slowed membrane trafficking in activated macrophages was a prolonged exposure of incoming molecules to an acidic nonlysosomal compartment, a condition which may facilitate microbicidal chemistries or antigen processing. PMID- 11037970 TI - Osteopontin augments CD3-mediated interferon-gamma and CD40 ligand expression by T cells, which results in IL-12 production from peripheral blood mononuclear cells. AB - Osteopontin is an RGD-containing bone matrix protein with cytokine-like functions that is associated with early stages of Th1-mediated diseases. Although the function of osteopontin in these responses is unknown, it is expressed by activated T cells and macrophages and can costimulate T cell proliferation. Studies have demonstrated that early IL-12 and IFN-gamma expression is required to induce a protective response to many intracellular pathogens. Herein, we demonstrate that osteopontin stimulation augments the ability of anti-CD3 monoclonal antibody to induce CD40 ligand (CD40L) and IFN-gamma expression on human T cells, resulting in CD40L- and IFN-gamma-dependent IL-12 production in vitro. These findings suggest a functional role for osteopontin in early Th1 responses, namely regulation of T cell-dependent IL-12 production. Further, osteopontin up-regulation of CD40L provides mechanistic support for the association of osteopontin with polyclonal B cell proliferation and humoral autoimmune disease. PMID- 11037971 TI - Immunopharmacological activity of Echinacea preparations following simulated digestion on murine macrophages and human peripheral blood mononuclear cells. AB - We have investigated the immunostimulatory, anti-inflammatory, and antioxidant activities of various Echinacea raw materials and commercially available products on murine macrophages and human peripheral blood mononuclear cells (PBMCs). To emulate oral dosing, a simulated digestion protocol was employed as a means of sample preparation. Echinacea-induced macrophage activation was used as a measure of immunostimulatory activity determined via quantitative assays for macrophage derived factors including tumor necrosis factor alpha, interleukin (IL)-1alpha, IL-1beta, IL-6, IL-10, and nitric oxide. Echinacea herb and root powders were found to stimulate murine macrophage cytokine secretion as well as to significantly enhance the viability and/or proliferation of human PBMCs in vitro. In contrast, Echinacea extracts chemically standardized to phenolic acid or echinacoside content and fresh pressed juice preparations were found to be inactive as immunostimulatory agents but did display, to varying degrees, anti inflammatory and antioxidant properties. PMID- 11037972 TI - Activation-dependent expression of the blood group-related lewis Y antigen on peripheral blood granulocytes. AB - The expression of the difucosyl-lactosamine type 2 oligosaccharide Lewis Y (LeY) on peripheral blood cells was investigated. As assessed by the reactivity with the mouse anti-LeY monoclonal antibody (mAb) ABL 364 among circulating blood cells, the expression of the LeY oligosaccharide was uniquely restricted to granulocytes. Although the density of LeY expressed on resting granulocytes was weak, in vitro activation of granulocytes with fMLP induced a rapid and pronounced increase in granulocyte LeY expression. Analysis of CEA-related glycoproteins immunoprecipitated with anti-CD66 mAbs followed by immunoblotting with mAb ABL 364 showed that granulocyte LeY is attached to members of the CD66 cluster, in particular to the 160/90 kD glycoprotein recognized by anti-CD66 mAb CBL/gran 10. The activation-associated increase in LeY attached to CD66 adhesion molecules implicates a role of the LeY determinant in the cytoadhesive properties of granulocytes. PMID- 11037973 TI - Bleomycin-induced pulmonary fibrosis is independent of eosinophils. AB - Eosinophils have been shown to increase in tissues during many fibrotic conditions and consequently have been suggested to contribute to the development of fibrosis. This study tested the hypothesis that eosinophils are essential in the development of lung fibrosis in mice in response to bleomycin (BLM). Anti-IL 5 antibody was administered intraperitoneally into mice 2 h prior to endotracheal BLM inoculation and thereafter, every other day. Lung eosinophilia was evaluated by measurement of eosinophil peroxidase activity and confirmed by eosinophil counts in histologic sections. Lung fibrosis was evaluated by hydroxyproline content and confirmed by collagen staining in histological sections. Results demonstrated that BLM induced pronounced lung eosinophilia, which was maximal 7 days after BLM treatment and remained elevated through day 14, in C57B1/6 SCID mice and CBA/J mice. In contrast, eosinophilia was a minor component in the lungs of wildtype C57B1/6 mice after BLM treatment, although lung fibrosis developed similarly in all three strains of mice. Treatment with anti-IL-5 completely abrogated eosinophilia but failed to block pulmonary fibrosis induced by BLM in all mouse strains, including C57B1/6 SCID, wildtype C57B1/6 mice, and CBA/J mice. Analysis of cytokine mRNA by RNase-protection assay in C57B1/6 SCID mice indicated that BLM treatment caused enhanced expression of the cytokines, TNF alpha, and IL-6 at days 3, 7, and 14 post-BLM inoculation, regardless of whether eosinophils were depleted by anti-IL-5. Finally, the importance of eosinophils in lung fibrosis was examined in IL-5 gene knockout mice (IL-5tm1Kopf). BLM treatment induced significant lung fibrosis in IL-5 knockout mice in the absence of eosinophilia. These findings indicate that eosinophils are not an absolute requirement for BLM-induced pulmonary fibrosis in the mouse. PMID- 11037974 TI - Escherichia coli cytotoxic necrotizing factor-1 (CNF-1) increases the adherence to epithelia and the oxidative burst of human polymorphonuclear leukocytes but decreases bacteria phagocytosis. AB - Recruitment of polymorphonuclear leukocytes (PMNL) is a hallmark of both urinary and digestive infections caused by Escherichia coli. Cytotoxic necrotizing factor 1 (CNF-1) is a toxin produced by uropathogenic E. coli strains that mediates its effects via the activation of small GTP-binding proteins. However, the role and the consequences of CNF-1 on PMNL physiology remain largely unknown. In this study, we provide evidence that CNF-1 dramatically affects the PMNL cytoskeleton architecture by inducing an increased content of F-actin. Furthermore, we demonstrate that CNF-1 increases functional features of PMNL, such as superoxide generation and adherence on epithelial T84 monolayers, but significantly decreases their phagocytic function. Our results suggest that CNF-1 may behave as a virulence factor in urinary or digestive infection by stimulating PMNL cytotoxicity as a result of its enhancing effect on their adherence to epithelial cells as well as the production of radical oxygen products. Moreover, the decreased phagocytosis of PMNL induced by CNF-1 likely facilitates growth of bacteria. In these conditions, CNF-1 would intervene in the initiation and in the perpetuation of the inflammatory process. PMID- 11037975 TI - Sequential migration of neutrophils across monolayers of endothelial and epithelial cells. AB - In the course of granulocyte-dominated lung inflammation, granulocytes migrate across the endothelium and epithelium of the lung and cause severe tissue damage. To study this process in more detail, we developed a bilayer transmigration model composed of primary human endothelial and lung epithelial cells, simultaneously cultured on opposite sides of Transwell filters. Electron microscopical analysis showed that the morphology of the cells and the expression of junctional proteins remained unaltered and that matrix components were deposited onto the filter. Intriguingly, neutrophil migration was more efficient across the bilayers than across single epithelial monolayers and did not differ from migration across single endothelial monolayers. Coculture experiments showed that endothelial cells stimulated epithelial cells to release IL-6 and that epithelial cells enhanced release of IL-8 from endothelial cells. Together these data reveal bidirectional signaling and enhanced neutrophil migration in a transmigration model of primary human epithelial and endothelial cells. PMID- 11037976 TI - A novel tumor necrosis factor (TNF) mimetic peptide prevents recrudescence of Mycobacterium bovis bacillus Calmette-Guerin (BCG) infection in CD4+ T cell depleted mice. AB - Tumor necrosis factor (TNF) is required to control mycobacterial infections, but its therapeutic value is limited by its in vivo instability and toxicity. The efficacy of a nontoxic TNF-mimetic peptide (TNF70-80) was tested in mice infected with Mycobacterium bovis bacillus Callette-Guerin (BCG). In vitro TNF70-80 and recombinant human TNF (hTNF) acted with interferon gamma (IFN-gamma) to reduce bacterial replication and to induce synthesis of bactericidal nitric oxide (NO) in BCG-infected, bone marrow-derived murine macrophages. The dose-dependent inhibitory effect on bacterial replication was blocked by neutralizing anti-IFN gamma and anti-hTNF mAbs. Further, n-monomethyl-L-arginine (n-MMA) and a soluble TNF-receptor I (TNFRI-IgG) blocked bacterial growth and NO synthesis. Therefore, the peptide acted with IFN-gamma via induction of NO synthase and signaled through TNFRI receptors. Concomitant in vivo treatment with TNF70-80 or hTNF prevented reactivation of chronic BCG infection in mice depleted of CD4+ T cells by injecting anti-CD4 antibodies. Granuloma number and bacterial load were comparable in treated, T cell-depleted mice and in chronically infected, intact animals. Thus, TNF70-80 and hTNF can modulate recrudescent BCG infection in CD4+ T cell-deficient mice. PMID- 11037977 TI - Monocyte:astrocyte interactions regulate MCP-1 expression in both cell types. AB - As astrocytes are a source of monocyte chemoattractant protein-1 (MCP-1) and lie in close apposition to brain microvessels, interactions between astrocytes and infiltrating monocytes might regulate production of this chemokine. To investigate this possibility, a monocyte:astrocyte co-culture model was utilized to assess the respective roles of these two cell types in regulating MCP-1 production. Results indicate that, while neither monocytes nor astrocytes alone produce detectable levels of MCP-1, co-culture of these two cell types results in time-dependent production of this chemokine. Such production requires de novo protein synthesis and is dependent on physical contact between monocytes and astrocytes, involving engagement of the cell-adhesion molecules ICAM-1 and VCAM 1. Additionally, interleukin 1-beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) are soluble mediators of this response. These findings imply that monocyte extravasation into the CNS may be critically regulated at the blood brain barrier by specialized monocyte:astrocyte interactions. PMID- 11037978 TI - Engagement of beta2 integrins induces surface expression of beta1 integrin receptors in human neutrophils. AB - Induction of beta1 integrin (CD49/CD29) expression in polymorphonuclear leukocytes (PMN) has been shown to be associated with transendothelial migration recently. Yet, beta1 integrin expression is relatively insensitive to cell activation with soluble agonists, such as N-formyl-methionyl-leucyl-phenylalanine (fMLP). We hypothesized that beta2 integrins (CD11/CD18), critically involved in PMN adhesion and extravasation, may play a role in regulating 1 integrin expression in PMN. Antibody cross-linking of CD18, mimicking adhesion-dependent engagement of beta2 integrins, resulted in rapid, tyrosine kinase-dependent upregulation of beta1 integrins. This response was potentiated by simultaneous chemoattractant (fMLP) stimulation of PMN. Moreover, upregulation of beta1 integrins evoked by CD18 cross-linking was found to support adhesion of fMLP stimulated PMN to matrix proteins and also was critical for the ability of PMN to migrate in collagen gels in response to a gradient of fMLP. Taken together, these data demonstrate that engagement of beta2 integrins in human PMN induces beta1 integrin expression in these cells of significance for their migration in the extravascular tissue. Thus, beta2 integrins may serve the function to regulate PMN locomotion in extravascular tissue via receptor crosstalk with beta1 integrins. PMID- 11037979 TI - CD97 isoform expression in leukocytes. AB - Different adhesive capacity in interactions with CD55 has been ascribed to the isoforms of the leukocyte CD97 antigen, CD97 (EGF 1,2,5), CD97 (EGF 1,2,3,5), and CD97 (EGF 1,2,3,4,5). In the study, coexpression of the three CD97 isoforms and predominance of CD97 (EGF 1,2,5) transcripts in leukocytes are demonstrated. The contribution of CD97 (EGF 1,2,3,5) and CD97 (EGF 1,2,3,4,5) to total CD97 levels varied among most cell types only slightly, although relatively higher mRNA levels of both isoforms were detected in U 937 cells and monocytes. In peripheral blood lymphocytes, CD97 isoforms did not show clear variation after PMA stimulation and were down-regulated equally after CD97 cross-linking. Moreover, the CD97 isoform pattern was not altered in monocytes after interferon-gamma stimulation and in synovial T cells from patients with rheumatoid arthritis. CD97 mRNA levels did not necessarily correspond to CD97 surface density. The findings suggest that adhesive activity of CD97 toward CD55 is unlikely to be regulated by differential CD97 isoform expression. PMID- 11037980 TI - Differential expression of the chemokine receptors by the Th1- and Th2-type effector populations within circulating CD4+ T cells. AB - The in vitro studies have proposed that human Th1 cells favor expression of CXCR3 or CCR5, whereas Th2 cells favor CCR3 and CCR4. In this study, the in vivo relevance of expression of these chemokine receptors on Th cells was investigated in patients with atopic dermatitis (AD) as the Th2-dominated disorder and nonatopic normal individuals. Flow-cytometric analysis using monoclonal antibodies against CXCR3, CCR5, CCR3, and CCR4 disclosed that a substantial proportion of memory (CD45RO+) CD4+ T cells in the blood of AD and normal patients expressed CXCR3, CCR5, or CCR4, but expression of CCR3 on these cells was negligible. Stimulation studies combined with intracellular cytokine staining revealed that the cells capable of producing Th2 cytokines, such as interleukin-4 (IL-4), IL-5, and IL-13, were restricted to the CCR4-expressing population within memory CD4+ T cells. Concerning Th1 cytokine production, interferon-gamma (IFN gamma)-producing cells resided exclusively in CXCR3-expressing memory CD4+ T cells, although IFN-gamma production was found in both memory CD4+ T cells with and without CCR5 expression. We observed that CCR4-expressing memory CD4+ T cells in the blood were more increased in AD patients as compared with normal patients, whereas CXCR3-expressing memory CD4+ T cells were present in a lower frequency in AD than seen in normal patients. These results suggest that CXCR3 and CCR4, but not CCR5 or CCR3, appear to serve as the useful markers for identification of circulating Th1 and Th2 effector populations. PMID- 11037981 TI - Differential localization of protein kinase C isotypes in equine eosinophils and neutrophils. AB - Phorbol esters, which activate protein kinase C (PKC), stimulate equine eosinophil superoxide production and adherence. After showing that superoxide production could be inhibited by the nonselective PKC inhibitors, staurosporine and bisindolymaleimide I, the PKC isotypes in equine eosinophils were characterized, because evidence suggests that individual isotypes may play distinct roles in regulating eosinophil function. Western blots demonstrated that equine eosinophils expressed PKC alpha, beta, delta, epsilon, iota, and zeta. However, unlike the equine neutrophil, the majority of the PKC was detected in the particulate fraction of the cell. Despite this unusual location, the PKC in equine eosinophils was activatable, suggesting that it is functionally competent. The regulatory role of PKC in equine eosinophils may reflect the association of activity with the particulate fraction and the profile of isotype expression. PMID- 11037983 TI - Nuclear Regulatory Commission revises medical policy statement. PMID- 11037982 TI - Recognition of CHO cells by inhibitory and activating Ly-49 receptors. AB - Upon ligand recognition, members of the murine Ly-49 receptor family can transmit inhibitory or activating signals that regulate NK cell function. Ly-49A, G, and D have been shown to recognize the murine class I molecule H-2Dd as a potential ligand. Recent studies also have demonstrated also that Ly-49D+ NK cells can lyse CHO cells, although the ligand responsible for this recognition was not identified. Because allorecognition by NK cells may be important in bone-marrow transplantation and because of the overlapping class I recognition by these receptors, recognition of CHO cells by Ly-49G and A was investigated. Our data suggest that Ly-49G and probably A transmit inhibitory signals in response to CHO cells. Receptor inhibition was assessed by examining NK lytic function, IFN-gamma secretion, and DAP12 phosphorylation in response to CHO cells by sorted subsets of Ly-49D vs. G B6 NK cells. Our results suggest that CHO cells may express a common ligand(s) that is capable of engaging Ly-49D, G, and possibly A in C576BL/6 NK cells. In addition to our findings that Ly-49 inhibitory receptors also recognize CHO cells, activating receptors other than Ly-49D are present in B6 mice that can lyse CHO cells. PMID- 11037984 TI - Tracking the origin of the NRC 30-mCi rule. PMID- 11037985 TI - William H. Beierwaltes: a life in nuclear medicine. PMID- 11037986 TI - 99mTc-sulfur colloid gastroesophageal scintigraphy with late lung imaging to evaluate patients with posterior laryngitis. AB - The aim of this study was to use gastroesophageal and pulmonary scintigraphy to evaluate the prevalence of gastroesophageal reflux and airway involvement among patients with posterior laryngitis. METHODS: The study included a total of 201 patients (131 females, 70 males; age range, 15-77 y; mean age +/- SD, 49 +/- 16 y). All patients had posterior laryngitis documented by laryngoscopy and symptoms such as a dry cough, painful swallowing, and hoarseness. A control population of 20 healthy volunteers (13 females, 7 males; age range, 19-74 y; mean age, 53 +/- 13 y) was also evaluated. After a 12-h fast, all subjects underwent gastroesophageal scintigraphy through administration of 300 mL orange juice labeled with 185 MBq 99mTc-sulfur colloid. After 18 h, planar anteroposterior thoracic images were acquired with the subjects supine. RESULTS: Sixty-seven percent of patients (134/201) had scans positive for gastroesophageal reflux; of these, 30 (22%) had distal reflux and 104 (78%) had proximal reflux. In addition, the scans of 31 patients were positive for proximal reflux-associated pulmonary uptake. The frequency, duration, and degree of reflux episodes were significantly greater in patients with proximal reflux than in patients with distal reflux (P < 0.001). The 67 patients in whom reflux was not detected had diseases or reflux associated cofactors that could account for laryngeal symptoms. No statistically significant difference in symptoms or esophageal motility parameters could be identified among the patient groups, but patients with proximal reflux had significantly prolonged gastric emptying times compared with healthy volunteers. CONCLUSION: Most patients with posterior laryngitis had detectable proximal gastroesophageal reflux. Exposure of the proximal part of the esophagus to acid, by setting the stage for microaspiration of gastric material into the larynx, remains a major cause of damage to the laryngeal mucosa. Slowed gastric emptying may be a predisposing factor. Moreover, symptoms such as a dry cough, painful swallowing, or hoarseness may not be reliable predictors of the presence of gastroesophageal reflux or of associated airway involvement. PMID- 11037987 TI - Pulmonary distribution and kinetics of inhaled [11C]triamcinolone acetonide. AB - Triamcinolone acetonide (TAA) is an anti-inflammatory steroid used for topical treatment of allergic rhinitis and asthma. Drug deposition onto target tissues is an important parameter, so methods for accurate deposition measurement are needed. Lung deposition is especially problematic to measure because of the large field of view and low relative drug penetration. Our main objective was to use PET to measure the deposition and postdeposition kinetics of TAA in the lung after administration from the Azmacort inhaler. The second objective was to evaluate changes in distribution caused by the inhalation spacer that is built into the product. METHODS: 11C-labeled TAA was formulated as the Azmacort product, 5 healthy volunteers inhaled it, and PET scans were obtained of its distribution in the head and chest. Region-of-interest analysis with CT overlay was used to analyze the distribution and kinetics in the airway and lung. RESULTS: From 10% to 15% of the inhaled drug dose was deposited in target airway regions in a distally decreasing pattern. Deposition in the oral cavity was about 30% of the dose. Slow absorption or clearance of drug from target tissues was observed over time. Use of the inhalation spacer caused statistically significant increases in all target tissues (factor of 2-5) and a roughly 40% decrease in oral deposition. Measurable amounts of the drug remained in target regions throughout the scanning period. CONCLUSION: Local pulmonary distribution and kinetics of inhaled drugs can be measured accurately by PET for drug development. The integrated actuator-spacer significantly enhanced deposition of TAA in target tissues and reduced deposition in the oropharyngeal region. PMID- 11037988 TI - Noninvasive estimation of FDG input function for quantification of cerebral metabolic rate of glucose: optimization and multicenter evaluation. AB - The aims of this study were to determine whether body weight or body surface area (BSA) should be used for noninvasive measurement of the cerebral metabolic rate of glucose (CMRGlc) by FDG PET with a standardized input function (SIF) and an autoradiographic method and to validate the procedure in a large population from different PET centers. METHODS: Plasma input functions measured by intermittent arterial blood sampling after intravenous injection of FDG, in 101 patients from 1 institution who were fasting for at least 4 h, were used to generate the SIF. The SIF was generated by averaging over 101 patients the input function normalized with the net injected dose and initial distribution volume (DV) of FDG estimated by the formula c x Ha x Wb, where H is body height and W is body weight. To evaluate the estimation of DV by BSA or body weight, the coefficient of variation (CV) of the ratio of Ha x Wb to the measured DV was calculated by changing a and b independently. Estimation of the CMRGlc with SIF based on the formula for DV was validated with an additional 192 subjects from 3 institutions who underwent FDG PET while fasting. The result of simulation was compared with the results of 4 previously published formulas for BSA and body weight. RESULTS: The optimal set of parameters, in which a was 0.80 and b was 0.35, minimized the CV. The averaged percentage error of the CMRGlc based on the optimal set of parameters for DV estimation and SIF was 8.9% for gray matter and 10.6% for white matter. Four BSA formulas brought about a similar error, which was significantly smaller than that based on body weight (P < 0.001, ANOVA). CONCLUSION: Noninvasive estimation of CMRGlc is made possible by careful measurement of the net injected dose and BSA. PMID- 11037989 TI - Does performing image registration and subtraction in ictal brain SPECT help localize neocortical seizures? AB - Ictal brain SPECT (IS) findings in neocortical epilepsy (patients without mesiotemporal sclerosis) can be subtle. This study is aimed at assessing how the seizure focus identification was improved by the inclusion of individual IS and interictal brain SPECT (ITS)-MRI image registration as well as performing IS - ITS image subtraction. METHODS: The study involved the posthoc analysis of 64 IS scans using 99mTc-ethyl cysteinate dimer that were obtained in 38 patients without mesiotemporal sclerosis but with or without other abnormalities on MRI. Radiotracer injection occurred during video-electroencephalographic (EEG) monitoring. Patients were injected 2-80 s (median time, 13 s) after clinical or EEG seizure onset. All patients had sufficient follow-up to correlate findings with the SPECT results. All patients had ITS and MRI, including a coronal volume sequence used for registration. Image registration (IS and ITS to MRI) was performed using automated software. After normalization, IS - ITS subtraction was performed. The IS, ITS, and subtraction studies were read by 2 experienced observers who were unaware of the clinical data and who assessed the presence and localization of an identifiable seizure focus before and after image registration and subtraction. Correlation was made with video-EEG (surface and invasive) and clinical and surgical follow-up. RESULTS: Probable or definite foci were identified in 38 (59%) studies in 33 (87%) patients. In 52% of the studies, the image registration aided localization, and in 58% the subtraction images contributed additional information. In 9%, the subtraction images confused the interpretation. In follow-up after surgery, intracranial EEG or video-EEG monitoring (or both) has confirmed close or reasonable localization in 28 (74%) patients. In 6 (16%) patients, SPECT indicated false seizure localization. CONCLUSION: Image registration and image subtraction improve the localization of neocortical seizure foci using IS, but close correlation with the original images is required. False localizations occur in a minority of patients. PMID- 11037991 TI - Reproducibility of renal length measurements with 99mTc-DMSA SPECT. AB - Renal length measurements are used in evaluating several abnormalities of the pediatric genitourinary tract. This study assesses reproducibility of renal length measurements obtained with 99mTc-dimercaptosuccinic acid (DMSA) SPECT. METHODS: The lengths of 98 kidneys of 51 children (age range, 1-16 y; mean age, 5.4 y) who underwent 99mTc-DMSA SPECT were measured independently by 2 observers. Renal length was calculated by converting pixels between points at the superior and inferior renal margins on a summated coronal image to centimeters. Lengths were measured for kidneys as they appeared in situ and after realignment along their long axes. SPECT reconstruction, choice of display parameters, positioning of points used for measuring, and alignment were performed independently by each observer. Interobserver variability, interobserver correlation, and mean differences between observers' measurements (expressed as measurement of observer 2 - measurement of observer 1) were calculated. RESULTS: Correlation between the observers' measurements was highly significant for both nonaligned and aligned studies (r = 0.95 and 0.97, respectively; both, P < 0.0001). Interobserver variability expressed as 1 SD was 3.6 mm for nonaligned studies and 2.8 mm for aligned studies. The mean difference between the 2 observers' measurements for nonaligned studies was 2.0 +/- 4.8 mm (P < 0.0001) with a range of -11 to 14 mm. For aligned studies the mean difference between the 2 observers' measurements was -0.1 +/- 4.0 mm (P = 0.88) with a range of -20 to 10 mm. Differences between observers were not dependent on absolute renal length (P = 0.68 for nonaligned studies; P = 0.40 for aligned studies). CONCLUSION: The variability in renal length measurements determined by 99mTc-DMSA SPECT is similar to that reported previously using sonography. Because the interobserver differences in renal length are similar to annual renal growth rates during childhood, caution should be applied when incorporating renal length measurements determined by 99mTc-DMSA SPECT into management algorithms. Additional studies are required to further establish interobserver variability, to assess intraobserver variability, and to evaluate means of improving standardization. PMID- 11037990 TI - Parenchymal mean transit time analysis of 99mTc-DTPA captopril renography. AB - Proposed renal hemodynamic mechanisms of captopril suggest that quantitation of renographic retention parameters should help identify patients suspected of having renovascular disease. The parenchymal mean transit time (MTT) is theoretically superior to other measures of retention, but data supporting its superiority are few. METHODS: Two groups of subjects were studied with diethylenetriamine pentaacetic acid (DTPA) baseline and captopril renography, one (n = 43) with demographically defined essential hypertension (group I) and the other (n = 60) with a high prevalence of renovascular disease (group II). Abnormal parenchymal MTT values were derived from the statistical confidence limits of group I data and then applied to group II subjects for comparison with angiographic results. RESULTS: Depending on the sensitivity of the threshold chosen, specificity varied, but the overall accuracy of baseline parenchymal MTT for renovascular hypertension detection ranged from 54% to 58%. Change in parenchymal MTT (post-captopril - pre-captopril) accuracy was 55%-61% and was not significantly different. Neither method improved on previously reported quantitative or qualitative criteria. Group II subjects had significantly worse renal function than did group I subjects, and 23% had nondiagnostic renograms. CONCLUSION: Parenchymal MTT analysis of DTPA captopril renography is not more accurate and offers no advantages compared with qualitative renography or with more commonly used renographic measures in our subjects. This may relate to the high prevalence of renal dysfunction in our population. In subjects with renal dysfunction, the low sensitivity and the trend toward low specificity of parenchymal MTT do not support its routine use for the evaluation of renovascular disease among patients suspected of having renovascular hypertension. PMID- 11037992 TI - Equilibration of 6-[18F]fluoro-L-m-tyrosine between plasma and erythrocytes. AB - Intracranial or intraventricular blood pools have been suggested as noninvasive sources of an input function for quantitative PET. These techniques measure the concentration of the tracer in whole blood, but the concentration in plasma depends on the equilibration of the tracer between plasma and erythrocytes. METHODS: FDG, 6-[18F]fluoro-L-m-tyrosine (FmT), or its major metabolite, 6 [18F]fluoro-3-hydroxyphenylacetic acid (FHPAA), was added to blood samples obtained from healthy fasting volunteers along with radioiodinated human serum albumin (RIHSA). Samples were incubated at 37 degrees C for times between 10 s and 2 h and then plunged into an ice bath and centrifuged. Whole blood and plasma were counted for 18F and 125I activities. The resulting time courses were fit to successively more complex models, evaluated using an F test. RESULTS: All radioactivity associated with RIHSA remained in the plasma, whereas FDG equilibrated instantaneously between plasma and erythrocytes. FmT took about 1 h to equilibrate between plasma and erythrocytes; this time course could be described by a single exponential with a half-life of 10 min. FHPAA equilibrated within the first 5 min of the study. CONCLUSION: Our results show that, unlike FDG, the partitioning of FmT between plasma and erythrocytes is a relatively slow process. We present an analytic correction that may be applied to the measured time course of radioactivity in whole blood to obtain the time course of the tracer in plasma. PMID- 11037993 TI - Radiographic and scintigraphic courses of union in cervical interbody fusion: hydroxyapatite grafts versus iliac bone autografts. AB - This study investigated the radiographic and scintigraphic courses of union in cervical interbody fusion using hydroxyapatite (HA) grafts or iliac bone autografts. METHODS: Twelve patients underwent both serial plain radiography and bone scintigraphy during the 12 mo after surgery. Serial plain radiographs were obtained every month until the end of the study period. Bone scintigrams with 99mTc-hydroxymethylene diphosphonate (HMDP) were obtained at 2 wk and at 1, 2, 3, and 6 mo. Uptake of 99mTc-HMDP in the graft was expressed as a ratio of the counts in the graft to those in the axis. RESULTS: In the HA graft group, the plain radiographs of all patients showed a radiolucent stripe that disappeared 7.3 +/- 1.5 (mean +/- SD) months after surgery. In the autograft group, a radiolucent stripe around the graft was not seen for any patient, and union was confirmed by follow-up radiographs within 6 mo after surgery. The serial changes in the 99mTc-HMDP uptake ratio showed no difference between the 2 groups. The 99mTc-HMDP uptake ratio peaked 1 mo after surgery and decreased rapidly to a plateau within 2 mo. CONCLUSION: In the HA graft group, despite the presence of a radiolucent stripe around the graft for more than 6 mo, the scintigraphic course of union was not different from that in the autograft group. The likelihood is that the presence of a radiolucent stripe around the HA graft in the early months after surgery is not always a sign of pseudoarthrosis. PMID- 11037994 TI - Ability of somatostatin receptor scintigraphy to identify patients with gastric carcinoids: a prospective study. AB - Gastric carcinoids are of increasing clinical concern because they may develop in hypergastrinemic states, especially with the increased chronic use of potent acid suppressants that can cause hypergastrinemia. However, gastric carcinoids are difficult to diagnose. Somatostatin receptor scintigraphy (SRS) has a high sensitivity and specificity for localizing carcinoids in other locations. The purpose of this study was to determine whether SRS could localize gastric carcinoids. METHODS: Two groups of patients with Zollinger-Ellison syndrome (ZES) with hypergastrinemia, each having a different increased risk of developing gastric carcinoids, were studied. One hundred sixty-two consecutive patients with ZES were studied prospectively, with 39 having multiple endocrine neoplasia, type 1 (MEN-1) (high increased risk), and 123 not having MEN-1 (low increased risk). Patients were admitted to the hospital initially and then yearly, undergoing SRS with SPECT, upper gastrointestinal endoscopy, and Jumbo Cup biopsies of any gastric abnormalities, as well as random biopsies of the gastric body. Tumor localization studies were also performed. Both the results of the routine SRS interpretation and the results of a masked review, with particular attention to the stomach of high risk MEN-1 patients, were correlated with the gastric biopsy results. RESULTS: Gastric SRS localization was positive in 19 (12%) of 162 patients. Sixteen patients had a gastric carcinoid, and 12 of these patients had SRS localization. The sensitivity of SRS in localizing a gastric carcinoid was 75%, with a specificity of 95%. Positive and negative predictive values were 63% and 97%, respectively. CONCLUSION: SRS is a noninvasive method that can identify patients with gastric carcinoids with a reasonable sensitivity and a high specificity. SRS should prove useful in the treatment of patients with hypergastrinemic states that have an increased incidence of gastric carcinoids. In patients with MEN-1, one must realize that localization in the upper abdomen on SRS may be caused by a gastric carcinoid and not a pancreatic endocrine tumor. PMID- 11037995 TI - FDG PET and immunoscintigraphy with 99mTc-labeled antibody fragments for detection of the recurrence of colorectal carcinoma. AB - The aim of this study was to compare FDG PET with a new monoclonal antibody-based imaging agent that comprises an anti-carcinoembryonic antigen (CEA) monoclonal antibody Fab' fragment directly labeled with 99mTc. METHODS: Twenty-eight patients who were previously treated for colorectal carcinoma and in whom recurrence was suspected were examined with FDG PET and immunoscintigraphy. The most common indications were an elevation of serum CEA (13 patients), suggestive lesions documented by CT (9 patients), sonography (4 patients), and severe constipation (2 patients). Planar imaging and SPECT were performed 4-6 h after intravenous injection of the new imaging agent. Whole-body PET was performed 45 60 min after intravenous injection of FDG. The findings were confirmed by conventional diagnostic modalities, surgery, and histology. RESULTS: Histology confirmed local tumor recurrence in 9 of 28 patients. Clinical follow-up or CT confirmed the presence of liver metastases in 9 patients and lymph node involvement, lung metastases, and bone metastases in 2 patients each. The new agent correctly detected 8 of 9 local recurrences, whereas FDG PET was able to detect all 9 cases and in 1 case was false-positive. Liver metastases were confirmed in 9 patients by FDG PET but in only 1 patient by the new agent. Two cases with lymph node metastases and 2 cases with lung metastases were correctly identified by FDG PET, but none were detected by the new agent. Finally, bone metastases were identified in 1 patient by FDG PET but not with the new agent, whereas bone marrow infiltration (n = 1) was diagnosed by both imaging modalities. CONCLUSION: These results indicate that FDG PET and 99mTc-labeled anti-CEA Fab' are suitable for the diagnosis of local recurrence of colorectal carcinoma but that FDG PET is clearly superior in the detection of distant metastases (liver, bone, and lung) and lymph node involvement. PMID- 11037996 TI - The diagnostic utility of the lognormal behavior of PET standardized uptake values in tumors. AB - A meta-analysis of data primarily from PET oncologic investigations using FDG PET was performed. Its purpose was to establish statistical features of the distributions of standardized uptake values (SUVs) as possible aids in the diagnostic process. METHODS: We obtained 1536 values of oncologic markers from patient studies of 40 investigations in the literature. Statistical parameters were tabulated for analysis. RESULTS: A significant observation is that, unlike skewed SUV histograms, log10SUV has Gaussian behavior, which is not uncommon for biologic quantities. This was found for SUVs of FDG and 2 amino acids as well as a few other cancer markers. A possible model for explaining this is proposed. For FDG, the SD sigma of the log10SUVs for an average cancer category was 0.23. Examining data within the framework of the model points to physiologic factors as dominating SUV variability rather than PET protocols. When data for a single cancer category were available from multiple institutions, averages, mean(SUV)s, disagree beyond chance expectations. Diagnostic utility suggestions include a universal linear relationship between sensitivity and severity, defined as SUV/mean(SUV), on semilogarithmic probability paper; a generic receiver-operating characteristic curve for all cancers; using [log10(mean(SUVmal)/mean(SUVnorm))] divided by (sigma(mal)2 + sigma(norm)2)(1/2) as a simple diagnostic effectiveness measure; and using Gaussian log10SUVs to avoid erroneous P values. CONCLUSION: Using the logarithms of markers, such as SUVs, several advantages stemming from their Gaussian nature can be achieved with benefits ensuing to the diagnostic process. PMID- 11037997 TI - Treatment of nonresectable hepatocellular carcinoma with intrahepatic 90Y microspheres. AB - Treatment for nonresectable hepatocellular carcinoma (HCC) is palliative. The relatively greater arteriolar density of hepatic tumors compared with normal liver suggests that intrahepatic arterial administration of 90Y-microspheres can be selectively deposited in tumor nodules and results in significantly greater radiation exposure to the tumor than external irradiation. The purpose of this study was to determine the proportion (frequency) and duration of response, survival, and toxicity after intrahepatic arterial injection of 90Y-microspheres in patients with HCC. METHODS: Patients with documented HCC, Eastern Cooperative Oncology Group performance status 0-3, adequate bone marrow, and hepatic and pulmonary function were eligible for study. Patients who had significant shunting of blood to the lungs or gastrointestinal (GI) tract or who could not undergo cannulation of the hepatic artery were excluded. Patients received a planned dose of 100 Gy through a catheter placed into the hepatic artery. RESULTS: Twenty-two patients were treated with 90Y-microspheres; 20 of the treated patients (median age, 62.5 y) were evaluated for treatment efficacy. Nine patients were Okuda stage I, and 11 were Okuda stage II. The median dose delivered was 104 Gy (range, 46-145 Gy). All 22 treated patients experienced at least 1 adverse event. Of the 31 (15%) serious adverse events, the most common were elevations in liver enzymes and bilirubin and upper GI ulceration. The response rate was 20%. The median duration of response was 127 wk; the median survival was 54 wk. Multivariable analysis suggested that a dose >104 Gy (P = 0.06), tumor-to-liver activity uptake ratio >2 (P = 0.06), and Okuda stage I (P = 0.07) were associated with longer survival. CONCLUSION: Significantly higher doses of radiation can be delivered to a HCC tumor by intrahepatic arterial administration of 90Y-microspheres than by external beam radiation. This treatment appears to be beneficial in nonresectable HCC with acceptable toxicity. PMID- 11037998 TI - Factors affecting sentinel node localization during preoperative breast lymphoscintigraphy. AB - Variable success rates for identifying axillary (AX) sentinel nodes in breast cancer patients using preoperative lymphoscintigraphy have been reported. We evaluated the effects of age, weight, breast size, method of biopsy, interval after biopsy, and imaging view on the success of sentinel node identification and on the kinetics of radiopharmaceutical migration. METHODS: Preoperative breast lymphoscintigraphy was performed in consecutive breast cancer patients from February 1998 to December 1998. The ipsilateral shoulder was elevated on a foam wedge and the arm was abducted and elevated overhead. Imaging using this modified oblique view of the axilla (MOVA) started immediately after peritumoral injection of Millipore-filtered 99mTc-sulfur colloid and continued until AX sentinel nodes were identified. Anterior views were obtained after MOVA. AX, internal mammary (IM), and clavicular (CL) basins were monitored in all patients. MOVA was compared with the anterior view for sentinel node identification. Age, weight, breast size, method of biopsy, interval after biopsy, and primary tumor location were evaluated for their effects on sentinel node localization and transit times from injection to arrival at the sentinel nodes. RESULTS: Seventy-six lymphoscintigrams were obtained for 75 patients. AX sentinel nodes were revealed in 75 (99%) cases. IM or CL sentinel nodes were found in 19 (25%) cases and were not related to tumor location; exclusive IM drainage was present in 1 (1%) case. Identification of AX sentinel nodes was equivalent with MOVA and anterior views in 18 (24%) patients, was better with MOVA in 20 (26%) patients, and was accomplished only with MOVA in 38 (50%) patients. Median transit time was 17.5 min (range, 1 min to 18 h) after injection, and larger breast size was associated with increased transit time. No effect of age, weight, biopsy method, interval from biopsy, or tumor location on transit time was found. CONCLUSION: Use of MOVA can improve identification of AX sentinel nodes. Although AX drainage is the predominant pattern, a tumor in any portion of the breast can drain to IM sentinel nodes. Transit time was influenced by breast size. Overall short arrival times with this technique allow sentinel lymph node dissection to be performed on the same day as lymphoscintigraphy. PMID- 11037999 TI - Evaluation of toxicity and efficacy of 186Re-hydroxyethylidene diphosphonate in patients with painful bone metastases of prostate or breast cancer. AB - Twenty-eight patients (12 men with prostate cancer, 16 women with breast cancer) were included in a phase II trial to evaluate the efficacy of 186Re hydroxyethylidene diphosphonate (HEDP) on pain from bone metastasis and the toxicity of this agent. METHODS: After intravenous administration of 1295 MBq 186Re-HEDP, the efficacy was evaluated by means of a daily log. RESULTS: We observed an objective response in 67% of prostate cancer patients and in 36% of breast cancer patients. The mean duration of response was 45 d for prostate cancer patients and 24 d for breast cancer patients. No major adverse effects were observed. Marrow toxicity did not exceed grade 2 for white blood cells and grade 3 for platelets using National Cancer Institute criteria. CONCLUSION: 186Re HEDP provides safe symptomatic relief of pain in prostate cancer patients. The benefit of this treatment is less clear in breast cancer patients. Further studies should be conducted to evaluate treatment by 186Re-HEDP at an earlier stage of the disease. PMID- 11038000 TI - Grading of tumors and tumorlike lesions of bone: evaluation by FDG PET. AB - Clinical diagnosis of skeletal tumors can be difficult, because such lesions compose a large, heterogeneous group of entities with different biologic behaviors. The aim of this prospective study was to assess the value of PET in grading tumors and tumorlike lesions of bone. METHODS: Two hundred two patients with suspected primary bone tumors were investigated using FDG PET. Uptake of FDG was evaluated semiquantitatively by determining the tumor-to-background ratio (T/B). All patients underwent biopsy, resulting in the histologic detection of 70 high-grade sarcomas, 21 low-grade sarcomas, 40 benign tumors, 47 tumorlike lesions, 6 osseous lymphomas, 6 plasmacytomas, and 12 metastases of an unknown primary tumor. RESULTS: All lesions, with the exception of 3 benign tumors, were detected by increased FDG uptake. Although sarcomas showed significantly higher T/Bs than did latent or active benign lesions (P < 0.001), aggressive benign lesions could not be distinguished from sarcomas. Using a T/B cutoff level for malignancy of 3.0, the sensitivity of FDG PET was 93.0%, the specificity was 66.7%, and the accuracy was 81.7%. CONCLUSION: FDG PET provides a promising tool for estimating the biologic activity of skeletal lesions, implicating consequences for the choice of surgical strategy. PMID- 11038001 TI - Is there a role for FDG PET in the diagnosis of musculoskeletal neoplasms? PMID- 11038002 TI - Peptide receptor imaging and therapy. AB - This article reviews the results of somatostatin receptor imaging (SRI) in patients with somatostatin receptor-positive neuroendocrine tumors, such as pituitary tumors, endocrine pancreatic tumors, carcinoids, gastrinomas, and paragangliomas, or other diseases in which somatostatin receptors may also be expressed, like sarcoidosis and autoimmune diseases. [(111)In-DTPA0]octreotide is a radiopharmaceutical that has great potential for helping visualize whether somatostatin receptor-positive tumors have recurred. The overall sensitivity of SRI to localize neuroendocrine tumors is high. In several neuroendocrine tumor types, inclusion of SRI in the localization or staging procedure may be very rewarding in terms of cost effectiveness, patient management, or quality of life. The value of SRI in patients with other tumors, such as breast cancer or malignant lymphomas, or in patients with granulomatous diseases has to be established. The application of radiolabeled peptides may be clinically useful in another way: after the injection of [(111)In-DTPA0]octreotide, surgeons can detect tumor localizations by a probe that is used during the operation. This may be of particular value if small tumors with a high receptor density are present (e.g., gastrinomas). As the success of peptide receptor scintigraphy for tumor visualization became clear, the next logical step was to try to label these peptides with radionuclides emitting alpha or beta particles, or Auger or conversion electrons, and to perform radiotherapy with these radiolabeled peptides. The results of the described studies with 90Y- and (111)In-labeled octreotide show that peptide receptor radionuclide therapy using radionuclides with appropriate particle ranges may become a new treatment modality. One might consider the use of radiolabeled somatostatin analogs first in an adjuvant setting after surgery of somatostatin receptor-positive tumors to eradicate occult metastases and second for cancer treatment at a later stage. PMID- 11038003 TI - Blocking catabolism with eniluracil enhances PET studies of 5-[18F]fluorouracil pharmacokinetics. AB - Noninvasive methods for measuring the pharmacokinetics of chemotherapeutic drugs such as 5-fluorouracil (FU) are needed for individualized optimization of treatment regimens. PET imaging of [18F]FU (PET/[18F]FU) is potentially useful in this context, but PET/[18F]FU is severely hampered by low tumor uptake of radiolabel and rapid catabolism of FU in vivo. Pretreatment with eniluracil (5 ethynyluracil) prevents catabolism of FU. Hypothesizing that suppression of catabolism would enhance PET/[18F]FU, we examined the effects of eniluracil on the short-term pharmacokinetics of the radiotracer. METHODS: Anesthetized rats bearing a subcutaneous rat colorectal tumor were given eniluracil or placebo and injected intravenously 1 h later with [18F]FU or [3H]FU. In the 18F studies, dynamic PET image sequences were obtained 0-2 h after injection. Tumors were excised and frozen at 2 h and then analyzed for labeled metabolites by high performance liquid chromatography. Biodistribution of radiolabel was determined by direct tissue assay. RESULTS: Eniluracil improved tumor visualization in PET images. With eniluracil, tumor standardized uptake values ([activity/g]/[injected activity/g body weight]) increased from 0.72 +/- 0.06 (mean +/- SEM; n = 6) to 1.57 +/- 0.20 (n = 12; P < 0.01), and tumor uptake increased by factors of 2 or more relative to plasma (P < 0.05) and bone, liver, and kidney (P < 0.01). Without eniluracil (n = 5), 57% +/- 4% of recovered radiolabel in tumor at 2 h was on catabolites, with the rest divided among FU (2% +/- 1%), anabolites of FU (38% +/- 7%), and unidentified peaks (4% +/- 2%). With eniluracil (n = 8), catabolites, FU, and anabolites comprised 2% +/- 1%, 41% +/- 5%, and 57% +/- 4%, respectively, of the recovered radiolabel in tumors. CONCLUSION: Eniluracil increased tumor accumulation of 18F relative to host tissues and fundamentally changed the biochemical significance of that accumulation. With catabolism suppressed, tumor radioactivity reflected the therapeutically relevant aspect of FU pharmacokinetics--namely, uptake and anabolic activation of the drug. With this approach, it may be feasible to measure the transport and anabolism of [18F]FU in tumors by kinetic modeling and PET. Such information may be useful in predicting and increasing tumor response to FU. PMID- 11038004 TI - Enzyme inhibition as an aid to simplify pharmacokinetic measurements? PMID- 11038005 TI - Preliminary evaluation of 15-[18F]fluoro-3-oxa-pentadecanoate as a PET tracer of hepatic fatty acid oxidation. AB - The liver is an important site of fat oxidation. Abnormalities of hepatic mitochondrial fatty acid oxidation (HMFAO) are associated with obesity, type II diabetes, alcoholic hepatitis, and nonalcoholic steatohepatitis. Noninvasive assessment of HMFAO by PET has been impeded by the lack of a specific radiotracer. METHODS: No-carrier-added 15-[18F]fluoro-3-oxa-pentadecanoate (FOP) was synthesized and evaluated in living rats and isolated rat livers. RESULTS: FOP showed high uptake and slow clearance of radioactivity from livers in living rats. Inhibition of HMFAO by pretreatment of fasting rats with the carnitine palmitoyltransferase-I (CPT-I) inhibitor reduced the liver-to-blood ratio by 64%. In isolated rat livers, perfused in normoxic (95% O2) and hypoxic (15% O2) conditions with glucose (5 mmol/L) and palmitate (0.15 mmol/L), the externally measured kinetics of FOP showed reversible binding in tissue. The kinetics were adequately fit by a catenary 2-compartment model for estimation of tracer distribution volumes (DVs). The DVs of both compartments were found to correlate with fractional palmitate oxidation rate (FPOR) in experiments in normoxic and hypoxic conditions. The correlation was particularly strong for the slower second compartment (DV2 [mL/g dry weight] = 34.1 FPOR [mL/min/g dry weight] - 0.7, r = 0.89). Relatively small levels of diffusible metabolites of FOP were formed in vivo and in isolated rat liver. CONCLUSION: The selective uptake of FOP by liver and the high sensitivity of hepatic FOP DV to changes of HMFAO with CPT-I inhibition and hypoxia suggests potential usefulness for the 3-oxa fatty acid analog in assessments of hepatic mitochondrial oxidation of exogenous fatty acids with PET. These data emphasize that further studies are required to clarify the intracellular disposition of FOP in the liver and test its validity as a tracer of HMFAO over a broad range of conditions. PMID- 11038006 TI - Histochemical correlates of (15)O-water-perfusable tissue fraction in experimental canine studies of old myocardial infarction. AB - A method has been proposed to quantitate the myocardial water-perfusable tissue fraction (PTF) in the area of hypoperfused asynergic segments using (15)O-water (H2(15)O) and PET. This study investigated the histochemical correlates of PTF (and perfusable tissue index, PTI) in a canine model of old myocardial infarction. METHODS: Myocardial infarction was produced in 12 mongrel dogs, and PET was performed 1 mo later, providing quantitative parametric images of PTF, regional myocardial blood flow (MBF), and extravascular density from H2(15)O, (15)O-carbon monoxide, and transmission datasets. At the end of scanning, the myocardium was sectioned, and the PET images were compared directly with the corresponding myocardial sections. RESULTS: The distribution of tissue necrosis identified by histochemical staining corresponded well with the defect in PTF but not in MBF. PTF agreed with the equilibrium images of myocardial H2(15)O distribution, obtained after injection of a large bolus of H2(15)O. The defect surface area identified on PTF agreed well quantitatively with the morphometric estimates of the surface area of myocardial infarction. PTI agreed with the absolute proportion of histochemically defined normal myocardium (0.87 +/- 0.09 and 0.83 +/- 0.08, respectively; P < 0.01). Both PTF and PTI decreased significantly in segments of myocardial infarction and showed a significant difference between the transmural and nontransmural myocardial infarction. CONCLUSION: The absolute mass and proportion of histochemically defined noninfarcted tissue may be quantitated with PTF and PTI in the area of myocardial infarction segments. PMID- 11038007 TI - Synthesis of [76Br]bromofluorodeoxyuridine and its validation with regard to uptake, DNA incorporation, and excretion modulation in rats. AB - This investigation aimed to validate 5-[76Br]bromo-2'-fluoro-2'-deoxyuridine (BFU) as a proliferation marker using PET. METHODS: Five megabecquerels 76Br-BFU were injected into the tail vein of Sprague-Dawley rats. At 6 or 16 h after injection, the rats were killed and the radioactivity concentration was measured in 6 different organs and blood. The fraction of radioactivity incorporated into DNA was determined for the spleen and small intestine. In parallel experiments, the animals were pretreated with hydroxyurea. In a few experiments, the urinary excretion of radioactivity was measured from administration of 76Br-BFU until 6 h. A sample of urine was analyzed with HPLC. In separate experiments, rats were given different doses of cimetidine, and the organ uptake and the fraction of radioactivity in DNA were determined at 24 h. RESULTS: The highest organ uptake of radioactivity was found in the spleen, followed by the small intestine. Approximately 90% of the radioactivity in these organs was incorporated into DNA, and inhibition by hydroxyurea was pronounced. Intact tracer constituted more than 95% of the radioactivity in urine. With cimetidine, the uptake of radioactivity increased approximately 2-5 times at different doses, whereas the urine radioactivity decreased markedly. CONCLUSION: 76Br-BFU was predominantly incorporated into DNA after administration in vivo in rats. If cimetidine was given in combination with the tracer, an increased contrast of radioactivity concentration between organs of high proliferation and organs of low proliferation was observed. The investigation suggested that 76Br-BFU has good potential as a PET tracer for the assessment of proliferation in vivo. PMID- 11038008 TI - Treatment of SW620 cells with Tomudex and oxaliplatin induces changes in 2-deoxy D-glucose incorporation associated with modifications in glucose transport. AB - Many studies suggest that changes in the uptake of the glucose analog FDG after therapy, compared with pretreatment uptake, predicts tumor response to therapy. However, clinical interpretation is compromised by a limited understanding of the effect of therapy on FDG and 2-deoxy-D-glucose (DG) uptake at the tumor cell level. METHODS: Uptake of 2-deoxy-D-[1-(3)H]glucose (3H-DG) by SW620 colonic tumor cells was measured before and 8, 16, 24, and 48 h after treatment with the novel platinum drug oxaliplatin and the novel thymidylate synthase inhibitor Tomudex. Glucose transport was determined by measuring the initial rate of uptake of the nearly nonmetabolized glucose analog 3-O-methyl-D-[1-(3)H]glucose (3H OMG). The effect of these drugs on cell cycle kinetics was determined using flow cytometry. RESULTS: Treatment of SW620 cells with oxaliplatin was found to decrease uptake of 3H-DG after up to 24 h, but uptake returned to control levels after longer treatment. The initial decrease in 3H-DG incorporation was associated with a lower rate of glucose transport. Treatment of cells with Tomudex induced an increase in 3H-DG uptake that depended on treatment duration. Both glucose transport and the volume of distribution of 3H-OMG were higher in Tomudex-treated cells than in control cells. Flow cytometry showed that oxaliplatin induced a G2 and M arrest, whereas a buildup of cells in the S phase was associated with Tomudex treatment. Both treatments induced apoptosis in SW620 cells. CONCLUSION: Changes in uptake of DG by SW620 colonic tumor cells responding to therapy is specific to the drug type. Modulation of glucose transport was associated with changes in 3H-DG uptake. PMID- 11038010 TI - 99mTc-apcitide scintigraphy and the detection of acute deep vein thrombosis. PMID- 11038009 TI - Accuracy of 131I tumor quantification in radioimmunotherapy using SPECT imaging with an ultra-high-energy collimator: Monte Carlo study. AB - Accuracy of 131I tumor quantification after radioimmunotherapy (RIT) was investigated for SPECT imaging with an ultra-high-energy (UHE) collimator designed for imaging 511-keV photons. METHODS: First, measurements and Monte Carlo simulations were carried out to compare the UHE collimator with a conventionally used, high-energy collimator. On the basis of this comparison, the UHE collimator was selected for this investigation, which was carried out by simulation of spherical tumors in a phantom. Reconstruction was by an expectation maximization algorithm that included scatter and attenuation correction. Keeping the tumor activity constant, simulations were carried out to assess how volume-of interest (VOI) counts vary with background activity, radius of rotation (ROR), tumor location, and size. The constant calibration factor for quantification was determined from VOI counts corresponding to a 3.63-cm-radius sphere of known activity. Tight VOIs corresponding to the physical size of the spheres or tumors were used. RESULTS: Use of the UHE collimator resulted in a large reduction in 131I penetration, which is especially significant in RIT where background uptake is high. With the UHE collimator, typical patient images showed an improvement in contrast. Considering the desired geometric events, sensitivity was reduced, but only by a factor of 1.6. Simulation results for a 3.63-cm-radius tumor showed that VOI counts vary with background, location, and ROR by less than 3.2%, 3%, and 5.3%, respectively. The variation with tumor size was more significant and was a function of the background. Good quantification accuracy (<6.5% error) was achieved when tumor size was the same as the sphere size used in the calibration, irrespective of the other parameters. For smaller tumors, activities were underestimated by up to -15% for the 2.88-cm-radius sphere, -23% for the 2.29-cm radius sphere, and -47% for the 1.68-cm-radius sphere. CONCLUSION: Reasonable accuracy can be achieved for VOI quantification of 131I using SPECT with an UHE collimator and a constant calibration factor. Difference in tumor size relative to the size of the calibration sphere had the biggest effect on accuracy, and recovery coefficients are needed to improve quantification of small tumors. PMID- 11038011 TI - Effects of various mutations in the neurophysin/glycopeptide portion of the vasopressin gene on vasopressin expression in vitro. AB - The vasopressin gene encodes three polypeptides besides the signal peptide: vasopressin, neurophysin II (neurophysin), and the carboxy-terminal glycopeptide (glycopeptide). Although the function of vasopressin is well characterized, those of the latter two are not completely understood. In the present study, we investigated the effects of various mutations within the neurophysin/glycopeptide portion of the vasopressin gene on vasopressin secretion in vitro, to clarify the role of each peptide in vasopressin biosynthesis. Expression vectors containing the vasopressin gene, either wild-type or various mutants, were transiently transfected into AtT20 cells, which are known to have the enzymes necessary for the proper processing of the vasopressin precursor protein. The amount of vasopressin secreted into the culture medium was estimated by specific radioimmunoassay. Variable degrees of decreased vasopressin secretion were observed with mutant vasopressin genes harboring deletions or amino acid substitutions in neurophysin. The naturally-occurring frame-shift mutation in the hereditary diabetes insipidus (Brattleboro) rat completely eliminated vasopressin expression. In contrast, a missense mutation found in patients with familial neurogenic diabetes insipidus only partially decreased vasopressin secretion. Finally, the mutant vasopressin gene lacking the N-linked glycosylation site in glycopeptide had no effect on vasopressin expression. Our data suggest that 1) intact neurophysin is not indispensable for vasopressin expression, although an altered structure of neurophysin significantly affects the secretion of the hormone; 2) the pathogenesis of diabetes insipidus with the two naturally occurring mutations found in the rat (Brattleboro rat) and human (familial central diabetes insipidus) seem to be different; and 3) glycosylation of the carboxy-terminal glycopeptide is not essential for the expression of vasopressin. PMID- 11038012 TI - Detection rates of TT virus DNA in serum of umbilical cord blood, breast milk and saliva. AB - To date, the routes of mother-to-infant transmission of TT virus (TTV) have not been fully elucidated. The present study examines the detection rates of TTV DNA in the serum of pregnant Japanese women and in cord blood at the time of delivery, as well as in the saliva and breast milk of mothers one-month postpartum. Primers derived from the well-known translated region N22 (N22 system), as well as the untranslated region (UTR system) were used. The prevalence of TTV DNA in the serum of pregnant women was found to be 11.9% (19/160) using the N22 system and 72.4% (55/76) using the UTR system. No TTV DNA was detected in the cord blood samples (0/160) when the N22 system was used for detection but TTV DNA was detected in 11.8% (7/76) of samples studied with the UTR system. Using the N22 system, TTV DNA was not detected in breast milk, but was detected in saliva. However using the UTR system, TTV DNA was detected in both specimens. These results imply that some babies are vertically infected with TTV via cord blood at the time of delivery or via breast milk or saliva. However, further research is necessary to confirm this hypothesis. polymerase chain reaction; pregnant women; horizontal route of transmission PMID- 11038013 TI - Prevention of nephrotoxicity of cisplatin by repeated oral administration of ebselen in rats. AB - The ability of ebselen, which exhibits glutathione peroxidase (GSH-Px)-like activity, to prevent cisplatin (CDDP)-induced nephrotoxicity was examined in rats. CDDP (6 mg/kg [20 micromol/kg] body weight) was injected intraperitoneally. In subgroups, daily ebselen doses of 2.75 (10 micromol), 5.5 (20 micromol), or 11.0 mg (40 micromol)/kg body weight were administrated orally 1 hour prior to CDDP treatment. Treatment with CDDP alone resulted in significantly increased plasma creatinine (Cr) and blood urea nitrogen (BUN) levels. Repeated administration of 5.5 and 11.0 mg/kg ebselen prevented the CDDP-induced elevation of plasma Cr and BUN levels and protected against kidney damage. Relative to controls, rat that received CDDP treatment displayed a decreased ratio of reduced glutathione (GSH) to oxidized glutathione (GSSG), an indicator directly related to oxidative stress, and elevated malondialdehyde (MDA) levels in the kidney. In comparison with controls, activity of GSH-Px activity, which antioxidant enzyme, was also reduced in the kidney of rats treated with CDDP. Repeated administration of 5.5 or 11.0 mg/kg ebselen prevented CDDP-induced alteration of GSH/GSSG ratios, MDA levels, and GSH-Px activity; however, no protection against CDDP was observed with administration of 2.75 mg/kg ebselen. Effective protection of CDDP induced nephrotoxicity with ebselen was observed only when the molar amount of each daily ebselen treatment equaled or exceeded PMID- 11038014 TI - Muller cells in the preconditioned retinal ischemic injury rat. AB - The role of Muller cells in the preconditioned retinal ischemic injury rat was investigated. In anesthetized Sprague Dawley rats, retinal ischemia for 5 minutes constituted the preconditioning stimulus for the left eye. After 24 hours, both eyes were clamped for 60 minutes. In 30, 60, 90, and 120, minutes and 1 day, 3 days, and 7 days after ischemia, electroretinograms were recorded, and the eyeballs were enucleated. After fixation with 4% paraformaldehyde, the avidin biotin-peroxidase technique was applied to show glutamine synthetase (GS) and glial fibrillary acidic protein (GFAP). Furthermore, for the solubilized retinas, Western blot analysis and enzyme-linked immunosorbent assay were performed to detect GS and GFAP in the extracts. Preconditioning performed 24 hours before ischemia significantly improved the recovery of the a-, and b-waves 1 day after 60 minute ischemia. In the 30, 60, 90, and 120 minutes after ischemia, the recovery of the a-wave only was observed. There was a nonsignificant trend toward greater recovery in the first 120 minutes after 60 minute ischemia, especially in the b-wave. GS immunoreactivity had no significant difference between non preconditioned and preconditioned groups 30, 60, 90, and 120 minutes after ischemia. In 1 day after ischemia, GS immunoreactivity decreased in both groups. In 3 and 7 days after ischemia, GS immunoreactivity recovered only in the preconditioned group. The retinas at 3 and 7 days after 1 hour of ischemia showed increased GFAP immunoreactivity in the non-preconditioned group. In the preconditioned group, only slight GFAP immunoreactivity was observed. These results suggested that the mechanism of preconditioned retinal ischemia may be related to Muller cells in the retina. PMID- 11038015 TI - TT virus infection in Japanese children: isolates from genotype 1 are overrepresented in patients with hepatic dysfunction of unknown etiology. AB - The pathogenecity of the TT virus (TTV) especially during childhood remains obscure. We investigated the prevalence of TTV in 40 patients with non-A to C hepatic dysfunction (non-A to C hepatic dysfunction group). Five patients with fulminant hepatitis of unknown etiology were enrolled in this group. We also examined 380 children without a history of transfusion or liver disease (control group). Subsequently, the genotypes of TTV strains isolated were analyzed in terms of their nucleotide sequences including 222 bp in the open reading frame 1 region. The prevalence of serum TTV DNA was 10/40 (25%) in the non-A to C hepatic dysfunction group and 25/380 (7%) in the control group. Sixty-six percent (23/35) of all examined cases exhibited either genotype 1 or 2. However, assessment of genotype in the non-A to C hepatic dysfunction group (10 cases) revealed a higher prevalence of genotype 1 than of all other genotypes (80% vs. 20%). This result differed significantly from that of the control group (25 cases; 32% vs. 68%). Such overrepresentation of genotype 1 suggests that this type of TTV strain is associated with the development of hepatic dysfunction of unknown etiology in Japanese children. PMID- 11038016 TI - Protective effect of melatonin on methylmercury-Induced mortality in mice. AB - Effect of melatonin on the mortality in methylmercury chloride (MMC)-intoxicated mice was evaluated. Mice were given MMC in the diet (40 mg Hg/g) with or without melatonin in drinking water (20 mg/ml) for 5 weeks. In the control group, given MMC alone, 4 of 10 mice began to show neurological signs (e.g., abnormal righting reflex, staggering gaitfallen and posture on its side) concomitant with loss of body weight 4-7 days before death. This group also showed 60% of survival rate on the 35th day. However, the treated group, concomitantly given melatonin, showed a 100% of survival rate on the 35th day, although 1 of 10 mice began to show the neurological signs on the 33rd day. The level of thiobarbituric acid reactive substance in the brain, as an indication of oxidative damage, showed a significant decrease in the treated group compared with the control group. Thus, the 100% survival rate in the treated group may be partly due to antioxidative effect of melatonin on the MMC induced neurotoxicity. PMID- 11038017 TI - Lamivudine as an alternative therapy for interferon-resistant chronic hepatitis B and the characteristics of hepatitis B virus: a case report. AB - A 27-year-old man who had been diagnosed as having chronic hepatitis B suffered disease exacerbation with marked reactivation of hepatitis B virus (HBV). Treatment with interferon (IFN) did not improve his condition, and his serum HBV DNA level increased to over 10 000 pg/ml during IFN administration. Following replacement with lamivudine, there was a substantial reduction in HBV DNA to an undetectable level, and liver function parameters subsequently improved to within the normal range. Quantitative analysis of the precore mutant HBV DNA, which is a variant that cannot express hepatitis B e antigen due to a G-to-A point mutation in the precore region of the viral genome, revealed that the amount present was greater than for the precore wild-type HBV DNA in the serum taken before IFN treatment. This case suggests that lamivudine would be an appropriate alternative to IFN, particularly in patients infected with HBV containing an excess of precore mutants resistant to IFN therapy. PMID- 11038018 TI - General decline of cancer incidence in US. PMID- 11038019 TI - France first in health care. PMID- 11038020 TI - France and Council of Europe against gene-patenting law. PMID- 11038021 TI - Task force on German fraud disillusioned. PMID- 11038022 TI - A row on euthanasia goes on while popular support increases. PMID- 11038023 TI - The regulation of tobacco and tobacco smoke. PMID- 11038024 TI - The optimal 5-fluorouracil regimen for the adjuvant therapy of colon cancer: where to from here? PMID- 11038025 TI - 'Hold the Mayo': solid facts or pulp fiction? PMID- 11038026 TI - QUOROM--another step forwards. PMID- 11038027 TI - QUOROM and the search for an updated 'clinical method' in the era of evidence based medicine. PMID- 11038028 TI - Therapeutic management of primary central nervous system lymphoma: lessons from prospective trials. AB - Primary central nervous system lymphomas (PCNSL) are aggressive malignancies, exhibiting one of the worst prognoses among lymphomas. The best treatment modality for PCNSL has not yet been identified. Several therapeutic questions still remain unanswered, and some methodological pitfalls in clinical trials prevent definitive conclusions from being drawn. In this review, certain aspects of trial design as well as emerging therapeutic guidelines are analyzed, and future perspectives are discussed. In the vast majority of prospective trials, general criteria for treatment of aggressive lymphomas were adopted, choosing primary chemotherapy (CHT) followed by radiotherapy (RT) as therapeutic modality. This strategy produced a five-year survival of 22%- 40% in comparison to the 3% 26% reported with RT alone. Systemic high-dose methotrexate (HD-MTX) seems to be the most effective drug, producing a response rate of 80%-90% and a two-year survival of 60%-65%. To date, the addition of other drugs at conventional doses have not consistently improved outcome. With a few exceptions, any regimen without HD-MTX comprehensively performed no better than RT alone. In combined treatment. RT doses should be decided on the bases of response to primary CHT and the number of lesions, and, until definitive conclusions from well-designed trials are available, RT parameters should follow the widely accepted principles used for other aggressive lymphomas. CHT as exclusive treatment, keeping RT for relapses or persistent disease, appears to be an attractive strategy. However, the worldwide experience with this modality is still limited, and corroborating data are needed. Intrathecal CHT still has not found a defined role in PCNSL management. Preliminary data seem to indicate that adequate meningeal treatment with HD-MTX, but without intrathecal CHT, could also be suitable in positive cerebrospinal fluid patients. Future efforts should be addressed to identify new active drugs and more efficient CHT combinations, to evaluate the efficacy of high-dose CHT supported by autologous peripheral blood stem cells transplantation, and to clarify the impact of RT delay in complete responders, the usefulness of intrathecal CHT, and the best management for elderly patients. The assessment of impact of treatment on neuropsychological functions and quality of life is a mandatory endpoint in clinical trials. PMID- 11038029 TI - Attitudes towards and participation in randomised clinical trials in oncology: a review of the literature. AB - BACKGROUND: Clinical trials are the principal means by which new treatment approaches are evaluated in medicine. It has been argued that randomised clinical trials provide the highest standard of care and at the same time help to contribute to scientific knowledge. However, only a relatively small proportion of cancer patients receive treatment as part of a formal clinical trial. This article provides a broad review of the issues pertinent to physician and patient participation in randomised clinical trials. METHODS: Search of computerised electronic databases (Medline, Psychlit, Cinhail, Embase). RESULTS: There are a variety of physician and patient characteristics which have previously been shown to influence participation in randomised clinical trials. Additionally, ethical concerns about randomised trials in general and the additional requirements of informed consent for clinical trials, may impact on recruitment. Whilst there is some research examining strategies to improve patient understanding about clinical trials and promote patient involvement in clinical decision-making, there are deficiencies in these areas. In particular there is a paucity of research examining the association between knowledge about clinical trials, anxiety associated with a new cancer diagnosis and willingness to participate in randomised clinical trials. CONCLUSIONS: Further research also is needed to evaluate strategies to better inform patients about clinical trials. PMID- 11038030 TI - Adjuvant chemotherapy with 5-fluorouracil, L-folinic acid and levamisole for patients with colorectal cancer: non-randomised comparison of weekly versus four weekly schedules--less pain, same gain. QUASAR Colorectal Cancer Study Group. AB - BACKGROUND: QUASAR is a large trial of adjuvant chemotherapy for colorectal cancer in which clinicians could choose to deliver a standard adjuvant cytotoxic chemotherapy regimen, 5-fluorouracil (5-FU) and L-folinic acid (L-FA), in either a once-weekly or a four-weekly schedule. We report results of a non-randomised comparison between these schedules with respect to survival, recurrence and differential toxicity. PATIENTS AND METHODS: In a factorial (2 x 2) trial design, QUASAR compared high-dose (175 mg) versus low-dose (25 mg) L-FA and levamisole versus placebo. The dose of 5-FU was fixed at 370 mg/m2 and although the recommended schedule was i.v. bolus delivery, daily for 5 days repeated four weekly for 6 months, a significant proportion of randomising clinicians were constrained to deliver once-weekly 5-FU-L-FA for 30 weeks. RESULTS: Four thousand nine hundred twenty-seven patients were entered into QUASAR between May 1994 and October 1997, eighteen hundred twenty-nine of whom have recurred and sixteen hundred eighty-nine died. Similar numbers 2370 vs. 2559 were treated with the once-weekly and four-weekly schedules and the demographic features of the 2 groups were well balanced: stage C, 73.3% once-weekly vs. 71.0% four-weekly; colon, 68.0% vs. 68.3%; high-dose FA, 50.1% vs. 49.9%; levamisole, 49.3% vs. 49.3%; females, 40.2% vs. 41.7%; median age (years) 62 vs. 61. The risk of recurrence and survival were similar regardless of schedule: three-year survival was 70.6% once-weekly vs. 71.0% four-weekly; three-year recurrence risk was 35.6% once-weekly vs. 35.5% four-weekly; But, the once-weekly regimen was much less toxic: number of patients for whom toxicity was reported (once-weekly: four weekly), stomatitis, 37 vs. 337; diarrhoea, 260 vs. 440; neutropenia, 20 vs. 153. CONCLUSIONS: The once-weekly regimen is much less toxic than and, apparently, about as effective as the four-weekly schedule. This suggests that the toxicity of 5-FU-L-FA adjuvant chemotherapy could be reduced substantially by weekly scheduling without compromising efficacy. Alternatively, efficacy might be enhanced with equal toxicity by more dose-intense weekly FU-L-FA regimens. However, this conclusion from a non-randomised comparison needs confirmation in prospective randomised studies. PMID- 11038031 TI - Trends in survival for patients diagnosed with cancer in Vaud, Switzerland, between 1974 and 1993. AB - BACKGROUND: Analysis of trends in cancer survival in defined well surveilled populations can provide useful indications on advancements in cancer management and treatment. PATIENTS AND METHODS: Survival rates from the Vaud Cancer Registry were computed for 31,158 cases registered in 1984-1993, and compared with those registered in 1974-1978 and 1979-1983. RESULTS: A systematic, albeit generally moderate, tendency towards increasing five-year relative survival was observed for both sexes and most major cancer sites, including oral cavity and pharynx (0.38-0.43). stomach (0.21-0.26), colon (0.49-0.55), rectum (0.45-0.51), lung (0.08-0.12), skin melanoma (0.67-0.89), female breast (0.67-0.80), endometrium (0.72-0.84), ovary (0.28-0.37). prostate (0.44-0.66), testis (0.73-0.96), bladder (0.31-0.50), kidney and renal pelvis (0.41-0.59), thyroid (0.73-0.81), non Hodgkin's lymphomas (0.37-0.63), Hodgkin's disease (0.61-0.81), and leukaemias (0.27-0.39). Survival for all cancers and both sexes combined, rose from 0.51 0.64 (0.57 for males, 0.71- for females). No appreciable change in survival was observed for cancers of oesophagus, liver, gallbladder, pancreas, larynx, cervix uteri, brain, multiple myeloma, as well as unidentified or unknown origin neoplasms. CONCLUSIONS: Survival estimates for most cancer sites are comparable to the US SEER dataset, and their pattern of trends are discussed in terms of improved diagnosis and treatment for various neoplasms. PMID- 11038033 TI - Cancer-related fatigue: inevitable, unimportant and untreatable? Results of a multi-centre patient survey. Cancer Fatigue Forum. AB - OBJECTIVES: To investigate cancer patients' experience of fatigue and their perceptions about the causes, management and impact of this symptom. DESIGN: Cross-sectional, questionnaire-based survey. SETTINGS: Three regional cancer centres; Glasgow, Birmingham and Southampton. PARTICIPANTS: One thousand three hundred seven outpatients with cancer attending the three units over a 30-day period. MAIN OUTCOME MEASURES: Investigator designed questionnaire and the fatigue sub-scale of the Functional Assessment of Cancer Therapy Fatigue (FACT-F) questionnaire. RESULTS: The response rate was 576 of 1307 (44%). Fatigue was reported to affect 58% of patients 'somewhat or very much'. The comparable figures for pain and nausea/vomiting were 22% and 18%, respectively. Fatigue had never been reported to the hospital doctor by 52% (281 of 538) of patients with this symptom. Only 75 patients (14%) had received treatment or advice about the management of their fatigue. Fatigue was reported to be not well-managed by 33% (180 of 538) of patients with this symptom. The comparable figures for pain and nausea/vomiting were 9% (46 of 538) and 7% (37 of 538), respectively. The median FACT-F score was 18 (range 0-52). On multivariate analysis 54% of the variation in FACT-F scores could be explained by the combination of quality of life, depression, dyspnoea, weight loss/anorexia and use of analgesics in the previous month. CONCLUSIONS: Fatigue has been identified as an important problem by patients with cancer. It affects more patients for more of the time than any other symptom and is regarded by patients as being more important than either pain or nausea/vomiting. Research into the aetiology and management of this symptom should be regarded as a priority. PMID- 11038032 TI - Skin tests predict survival after autologous tumor cell vaccination in metastatic melanoma: experience in 81 patients. AB - BACKGROUND: Currently there is no standard adjuvant treatment following surgical resection of metastatic melanoma. We investigated whether surgery followed by autologous tumor cell-BCG vaccination was beneficial for malignant melanoma patients. In this study we focus on the prognostic value of DTH response following vaccination therapy. PATIENTS AND METHODS: Eighty-one patients with AJCC stage III and IV melanoma were selected. Whenever feasible, radical metastasectomy was performed. ASI was initiated by the administration of three weekly intra-cutaneous vaccinations with 10(7) irradiated autologous tumor cells, starting four weeks after surgery. Depending on the size of DTH response to the first three injections, subsequent vaccinations were planned. The first two vaccines also contained 10(7) BCG organisms as an immune stimulatory adjuvant. RESULTS: Induration as well as erythema correlated strongly with survival (P < 0.0001 and P = 0.0004). After radical metastasectomy in stage III melanoma patients a five-year survival of 48% was observed. In stage IV disease, a five year survival of 34% was seen, after radical surgery had been performed. When macroscopic disease was present at start of vaccination treatment, no clinical responses occurred. Apart from transient skin ulceration at the site of BCG containing vaccinations, no serious side effects were observed. CONCLUSIONS: This study shows that large-scale preparation of autologous melanoma cell vaccines is feasible. while vaccination results in DTH responses that correlate significantly with survival. ASI seemed to be beneficial in stage III and stage IV melanoma when given in the adjuvant setting, while causing only very mild side effects. PMID- 11038034 TI - Clinical and pharmacological phase I study with accelerated titration design of a daily times five schedule of BBR3464, a novel cationic triplatinum complex. AB - OBJECTIVES: To define the maximum tolerated dose (MTD), the toxicity and pharmacokinetic profile of BBR3464, a novel triplatinum complex. PATIENTS AND METHODS: Fourteen patients with advanced solid tumors not responsive to previous antitumor treatments received BBR 3464 on a daily x 5 schedule every twenty eighth day. The drug was given as a one-hour infusion with pre-and post-treatment hydration (500 ml in one hour) and no antiemetic prophylaxis. The starting dose was 0.03 mg/m2/day. A modified accelerated titration escalation design was used. Total and free platinum (Pt) concentrations in plasma and urine were assessed by ICP-MS on days 1 and 5 of the first cycle. RESULTS: Dose was escalated four times up to 0.17 mg/m2/day. Short-lasting neutropenia and diarrhea of late onset were dose-limiting and defined the MTD at 0.12 mg/m2. Nausea and vomiting were rare, neither neuro- nor renal toxic effects were observed. BBR3464 showed a rapid distribution phase of 1 hour and a terminal half-life of several days. At 0.17 mg/m2 plasma Cmax and AUC on day 5 were higher than on day 1, indicating drug accumulation. Approximately 10% of the equivalent dose of BBR3464 (2.2%-13.4%) was recovered in a 24-hour urine collection. CONCLUSIONS: The higher than expected incidence of neutropenia and GI toxicity might be related to the prolonged half-life and accumulation of total and free Pt after daily administrations. Lack of nephrotoxicity and the low urinary excretion support the use of the drug without hydration. The single intermittent schedule has been selected for clinical development. PMID- 11038035 TI - Epidoxorubicin and docetaxel as first-line chemotherapy in patients with advanced breast cancer: a multicentric phase I-II study. AB - BACKGROUND: The combination of anthracyclines and taxanes is currently considered the first choice chemotherapy in advanced breast cancer (ABC) and considerable emphasis has been placed on programs exploring the safest and most efficient way to integrate these classes of drugs in both the metastatic and, more recently, the adjuvant setting. We report here the overall results of the combination of epidoxorubicin (E) 90 mg/m2 and docetaxel (D) 75 mg/m2 as first-line chemotherapy in ABC. PATIENTS AND METHODS: A total of 70 patients were entered in the initial dose-finding study (20 patients) and in the subsequent extended phase II trial (50 patients). Overall 54% of patients had dominant visceral disease and 57% had at least two metastatic sites. Adjuvant anthracyclines were allowed in the phase II part of the study based on the lack of cardiac toxicity observed in the phase I study at a median cumulative E dose of 480 mg/m2. A maximum of eight cycles of the combination was allowed, and cardiac function was monitored at baseline and after every second course by echocardiography. RESULTS: Overall, the median number of cycles administered with the combination was 4 (range 3-8). Neutropenia was confirmed to be the main haematological toxicity, with granulocyte colony stimulating factor (G-CSF) support required in 44% of the cycles. Febrile neutropenia occurred in 12% of cycles of the combination but 52% of the episodes could be managed on an outpatient basis with oral antibiotics. Overall, the median cumulative dose of E, including prior adjuvant anthracyclines, was 495 mg/m2 (range 270-1020 mg/m2). One patient who received adjuvant E together with radiotherapy to the left chest wall developed fully reversible clinical signs of cardiotoxicity and a significant decrease of LVEF to 35% after a cumulative E dose of 870 mg/m2, with four additional patients (6%) developing asymptomatic and transient decline of resting LVEF. The overall response rate (ORR) in 68 evaluable patients was 66% (95% confidence interval (95% CI): 54%-73%). A comparable antitumour activity of 71% was reported in the group of patients with a prior adjuvant chemotherapy with anthracyclines. After an overall median follow up time of 22 months (range 4-39+), the median time to progression (TTP) was 4.5 months and the median duration of response was 8 months (range 3-16). No pharmacokinetic (Pk) interaction could be demonstrated between E and D when given simultaneously and sequentially with a one-hour interval. CONCLUSIONS: The combination of E and D in a multiinstitutional setting is an active and safe regimen in poor-prognosis patients with ABC. New combinations and schedules are worth considering in an attempt to further improve disease response and long-term control of the disease. PMID- 11038036 TI - Vinorelbine-gemcitabine in advanced non-small-cell lung cancer (NSCLC): an AASLC phase II trial. Austrian Association for the Study of Lung Cancer. AB - PURPOSE: The purpose of the present phase 11 trial was to determine the efficacy and toxicity of vinorelbine-gemcitabine in patients with advanced non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: From December 1997 to February 1999, 78 chemotherapy-naive patients (median age 60 years, Karnofsky performance status of 100, 90, 80 and 70 present in 5%, 41%, 36% and 18% of the patients, respectively) with stage IIIB (17%) or IV (83%) NSCLC (65% adenocarcinomas, 22% squamous-cell carcinomas, 10% large-cell carcinomas, 3% mixed-cell carcinomas) received 25 mg/m2 vinorelbine and 1200 mg/m2 gemcitabine on days 1, 8 and 15 of a four-week cycle. RESULTS: In an intent-to-treat analysis, partial responses were seen in 19% of the patients. The median duration of response was 4.4 months. The median survival time was seven months and the one-year survival rate was 32%. Myelosuppression was the main side effect with WHO grade 3/4 neutropenia and thrombocytopenia in 35% and 11% of the patients, respectively. Other side effects were usually mild to moderate. CONCLUSIONS: Vinorelbine-gemcitabine is active, well tolerated and easy to administer on an outpatient basis in advanced NSCLC. Thus a randomized comparison of this combination with platinum-based protocols is warranted in patients with advanced NSCLC. PMID- 11038037 TI - Comparison of allogeneic transplant versus chemotherapy for relapsed childhood acute lymphoblastic leukaemia in the MRC UKALL R1 trial. MRC Childhood Leukaemia Working Party. AB - BACKGROUND: Although reinduction rates are good for children with relapsed acute lymphoblastic leukaemia there is no consensus on whether bone marrow transplantation (BMT) is the most effective treatment to prolong second remission. PATIENTS AND METHODS: Analyses comparing the outcome of related donor allogeneic BMT (related allograft) with chemotherapy are unreliable because of selection biases. To avoid these biases, the MRC UKALL R1 trial was analysed by HLA-matched donor availability. RESULTS: No significant difference in outcome was found between the donor and no donor groups. The donor group had a non significant eight-year event-free survival (EFS) advantage of 8%, (95% confidence interval -9%-24%) over the no donor group. Patients with a first remission less than two years appeared to benefit most from having a donor, although the effect was only marginally significantly different from patients with longer first remission. Analysis by treatment received gave similar results, with BMT patients having a 5% (P = 0.8) eight-year EFS advantage over patients who received chemotherapy. CONCLUSIONS: Related allograft was not found to be significantly better than chemotherapy, but there was the possibility of a moderate EFS benefit with related allograft. especially in patients with a short first remission. PMID- 11038038 TI - High-dose treatment with autologous bone marrow support as consolidation of first remission in younger patients with acute myelogenous leukaemia. AB - BACKGROUND: Debate and controversy remain as to the optimal post-remission therapy for younger patients with acute myelogenous leukaemia (AML). The aim of this study was to evaluate high-dose treatment (HDT) with autologous bone marrow support (ABMS) as consolidation of first complete remission (CR). PATIENTS AND METHODS: One hundred forty-four patients (AML-M3 excluded, median age 38 years, range 15-49 years) received remission induction therapy comprising: adriamycin 25 mg/m2, days 1-3, cytosine arabinoside (ara-C) and 6-thioguanine, both at 100 mg/m2 bid, days 1-7. Patients in whom CR was achieved received two further cycles of the same treatment prior to bone marrow being harvested and cryopreserved. HDT comprised ara-C: 1 g/m2 b.i.d. x six days and total body irradiation (TBI): 200 cGy b.i.d. for three days. Thawed autologous marrow was then re-infused. RESULTS: Complete remission was achieved in 106 of 144 patients (73%) who were thus eligible to receive ara-C + TBI + ABMS; 61 actually received it. Following HDT, the median time to neutrophil recovery (> 0.5 x 10(9)/l) was 25 days (range 11-72 days) and to platelet recovery (> 20 x 10(9)/l), 42 days (range 15-159 days). There were eight treatment-related deaths. Analysis by 'intention to treat' shows both remission duration (log-rank, P = 0.001) and survival (log-rank, P = 0.004) to be significantly longer for the 106 patients eligible to receive HDT than for a historical control group (n = 133) who received identical remission induction and consolidation therapy but without ara-C + TBI + ABMS. With a median follow-up of 5.5 years, 39 of 106 patients remain in CR (37%) and 54 (51% of those in whom CR was achieved) remain alive, with a predicted actuarial survival of 52% at 5 years. CONCLUSIONS: The addition of ara-C + TBI + ABMS to conventional consolidation therapy significantly improved remission duration and survival over those of a historical control group of patients with AML (aged < 50, AML-M3 excluded). HDT was, however, associated with significant treatment-related mortality and slow blood count recovery. The use of ara-C + TBI supported by peripheral blood progenitor cells should make the treatment safer and more widely applicable in AML. PMID- 11038039 TI - Tumor response and estrogen suppression in breast cancer patients treated with aromatase inhibitors. AB - BACKGROUND: The rationale for the hormonal treatment of breast cancer (BC) is based on depriving tumor cells of estrogenic stimulation. Aromatase inhibitors (Als) block the conversion of peripheral tissue androgens to estrogens with different levels of potency. In an attempt to investigate the relationship between tumor response and estrogen suppression, we reviewed the hormonal and clinical data of two previous studies with formestane (250 and 500 mg i.m. fortnightly) in advanced BC patients. PATIENTS AND METHODS: Two hundred four BC patients were selected on the basis of the availability of records concerning their plasma estrone (El) and estradiol (E2) levels assessed at scheduled times. The degree of estrogen suppression and the best clinical response of each patient during the trials were considered. RESULTS: There was a positive and significant (P < 0.05) correlation between baseline and post-formestane E1 and E2 levels, with a decrease in the levels of both hormones irrespective of any antitumor response. In particular, the degree of plasma estrogen suppression was similar in the patients who experienced a complete remission and those with progressive disease (PD). CONCLUSIONS: The plasma estrogen suppression induced by aromatase inhibition is not the only mechanism accounting for its clinical activity. Many clinical trials have demonstrated that all AIs induce a similar antitumor response regardless of their potency, and further investigations are warranted in order to improve our understanding as to why the patients with PD also show a significant plasma estrogen suppression. It is possible that intratumoral aromatase activity may be a marker for selecting the BC patients most likely to respond to AI treatment. PMID- 11038040 TI - Raltitrexed ('Tomudex') and radiotherapy can be combined as postoperative treatment for rectal cancer. AB - BACKGROUND: The optimal adjuvant therapy for operable rectal cancer is likely to be a combination of radiotherapy and chemotherapy. Raltitrexed ('Tomudex') is a specific thymidylate synthase inhibitor with a convenient administration schedule, acceptable and manageable toxicity, radiosensitising properties, and proven efficacy in the treatment of advanced colorectal cancer. It may, therefore, offer advantages compared with standard 5-FU chemotherapy regimens used in colorectal cancer. The aim of this phase I, dose-escalation study was to determine the recommended dose of raltitrexed for use with postoperative pelvic radiotherapy in patients with rectal cancer. PATIENTS AND METHODS: Patients with resected Dukes' stage B or C rectal cancer were treated with a combination of raltitrexed and radiotherapy (50.4 Gy at 1.8 Gy per fraction over five to six weeks). At least three patients were treated at each of three escalating raltitrexed dose levels (2.0, 2.6 and 3.0 mg/m2) once every three weeks. Toxicity was assessed by the recording of WHO adverse events and biochemistry and haematology determinations. RESULTS: A total of 22 patients entered the study, 17 of whom had Dukes' stage C disease. All three patients entered at a dose level of 3.0 mg/m2 experienced dose-limiting toxicity (DLT) (2 patients had grade 3 leucopenia and 1 patient had grade 2 leucopenia and grade 3 diarrhoea); however, only 2 of 1 patients entered at a dose level of 2.6 mg/m2 experienced DLT (1 patient had grade 4 neutropenia and 1 patient died probably due to aspiration pneumonia unrelated to treatment). The most common haematological toxic events were leucopenia (8 patients) and anaemia (6 patients). Only four haematological or biochemical toxic events were of grade 3 or 4. Other common toxicities were diarrhoea and nausea, which occurred in 15 and 9 patients, respectively. CONCLUSIONS: This study demonstrates that raltitrexed can be combined with postoperative radiotherapy for treatment of patients with Dukes' stage B or C rectal cancer. The recommended dose of raltitrexed in this setting is 2.6 mg/m2, which is close to the full monotherapy dose. PMID- 11038041 TI - Pegylated liposomal doxorubicin (doxil): reduced clinical cardiotoxicity in patients reaching or exceeding cumulative doses of 500 mg/m2. AB - BACKGROUND: The indications for pegylated liposomal doxorubicin (doxil) are expanding. We, therefore, wished to assess the safety of delivering doses exceeding 500 mg/m2 of doxil to patients with solid tumors. PATIENTS AND METHODS: Subjects accrued to eight phase I and II protocol studies conducted at two institutions, were assessed for cardiac function at baseline and at specified intervals by MUGA scans. In this retrospective analysis, the findings of 42 patients, from the total of 237 entered, who had reached or exceeded cumulative doses of 500 mg/m2 (range 500-1500 mg/m2) were reviewed. Changes in left ventricular ejection fraction (LVEF), and in clinical cardiac status were analyzed. Six patients, three who had received prior doxorubicin, also underwent endomyocardial biopsies after cumulative doses of 490-1320 mg/m2. RESULTS: None of the 42 patients had clinical congestive heart failure (CHF) secondary to cardiomyopathy. Post doxil MUGA scans were available for 41 of the 42 patients. Five had a drop of 10% or more in LVEF; three of these had received prior doxorubicin. Billingham endomyocardial biopsy scores ranged from 0-1 in five patients, while the sixth had a score of 1.5 after both 900 mg/m2 and 1320 mg/m2 doxil. Of a remaining 195 patients, 1 episode of CHF was recorded in a patient who had received 312 mg/m2 doxil over 120 mg/m2 of mitoxantrone and chest radiation. CONCLUSIONS: Cumulative doses in excess of 500 mg/m2 of doxil appear to carry a considerably lesser risk of cardiomyopathy as judged by serial LVEF's and clinical follow-up, than is generally associated with free doxorubicin. Heart biopsies have provided reassuring data in a small number of patients, even if pretreated with doxorubicin. However, since three doxorubicin pretreated patients were among the five experiencing drops in LVEF, more data are warranted on such patients. PMID- 11038042 TI - A phase II study of high-dose epirubicin in ovarian cancer patients previously treated with cisplatin. EORTC Gynecological Cancer Cooperative Group. AB - BACKGROUND: In vitro data demonstrated a dose-response relationship for doxorubicin in ovarian cancer cell lines. However, this dose-response question for anthracyclines has never been adequately addressed in ovarian cancer patients. A phase I study with epirubicin gave support to these in vitro findings and recommended a dose of 150 mg/m2 for phase II testing. PATIENTS AND METHODS: The present report concerns the final analysis of an EORTC-Gynecological Cancer Cooperative Group (GCCG) phase II study of high-dose epirubicin (HDE) in cisplatin-pretreated patients with epithelial ovarian cancer. A total of 100 eligible patients were included; 34 had progressed during first-line therapy (group 1), 17 had persistent disease after first-line therapy (group 2) and 49 had relapsed following an initial response to first-line therapy (group 3). All patients had measurable or evaluable disease, were aged < 75 years, had a WHO performance status 0-2, had adequate vital organ function and gave consent. Epirubicin was administered by rapid i.v. infusion at a dose of 150 mg/m2 and given at three-week intervals. Escalation to 180 mg/m2 was to be carried out if white blood cell nadir count was > 2.0 x 10(9)/l and platelet nadir count was > 75 x 10(9)/l. RESULTS: A total of 361 HDE treatment cycles were administered, the median number per patient being 4. Of the 85 patients who received at least two cycles of protocol treatment, 26 (31%) did not have any dose modification, 23 (26%) had dose reduction, while 36 (43%) had the dose increased to 180 mg/m2, at least for one cycle. The response rate in all eligible patients was 20% (95% confidence interval 13%-30%), 15% in group 1, 12% in group 2 and 27% in group 3. Patients with a cisplatin-free interval of > 12 months responded in 41%. The median duration of response was nine months (range 19 weeks to 3 years). Main toxicities were myelosuppression (leucopenia, neutropenia), nausea, vomiting, alopecia and mucositis. There were three cases of excessive toxicity leading to early discontinuation of HDE treatment and in one patient this contributed to death. No serious cardiotoxicity was recorded. CONCLUSIONS: It is concluded that HDE is active in platinum-pretreated patients with epithelial ovarian cancer and should be further studied in first-line in combination with paclitaxel and a platinum compound. PMID- 11038043 TI - Combination chemotherapy with navelbine and continuous infusion of 5-fluorouracil in metastatic, chemotherapy refractory breast cancer. AB - BACKGROUND: The protracted continuous infusion (PCI) of 5-fluorouracil (5-FU) has proven in several studies an active and well tolerated treatment for advanced, pretreated breast cancer. Navelbine has also activity in this setting. PATIENTS AND METHODS: Heavily pretreated patients with metastatic breast carcinoma were eligible for the study. Treatment consisted of 5-FU 250 mg/m2 given as a PCI by an elastomeric pump and navelbine 20 mg/m2 on days 1 and 8, every four weeks. Eighty-three patients (median age 54 years; range 32-82 years) entered the study. The median number of metastatic tumour sites was 2, with visceral involvement in 56 patients. Apart from five patients with contraindications, all patients had been pretreated with anthracyclines. Thirty-one patients had received taxanes and seventy-four bolus 5-FU. RESULTS: A median of 5 cycles (range 1-14) per patient was administered. The median duration of 5-FU infusion was 17 weeks (range, 4 90). In the 80 evaluable patients (3 not yet evaluable) 12 complete remissions and 24 partial remissions occurred (response rate, 45%). Median duration of response was 9 months. Toxicity was mild. Median survival was 20 months. CONCLUSIONS: PCI-5-FU combined with navelbine offers a reasonable chance of tumour regression with modest side effects in patients with heavily pretreated breast cancer. PMID- 11038044 TI - 5-Fluorouracil and folinic acid with or without CPT-11 in advanced colorectal cancer patients: a multicenter randomised phase II study of the Southern Italy Oncology Group. AB - PURPOSE: The combination regimen CPT-11 plus bolus and infusion 5-fluorouracil (5 FU) with high-dose leucovorin (hybrid regimen LV5FU2) has been tested for activity and toxicity against advanced colorectal carcinoma in a randomised, multicenter phase II trial. PATIENTS AND METHODS: A total of 102 chemotherapy naive patients were randomised in a 1:2 fashion to receive: leucovorin 100 mg/m2 administered as a two-hour infusion before 5-FU 400 mg/m2 as an intravenous bolus, and FU 600 mg/m2 as a 22-hour infusion immediately after 5-FU bolus injection repeated on days 1 and 2 (LV5FU2 regimen, arm A, 34 patients) or CPT-11 at 180 mg/m2 (150 mg/m2 for patients of age > or = 70 and < 75 years) only on day 1 immediately before LV5FU2 therapy (LV5FU2 + CPT-11 regimen, arm B 68 patients). Both treatments were repeated every two weeks. The presence of a calibration arm assured consistency and more realistic evaluation of results achieved with the LV5FU2 + CTP-II regimen. RESULTS: Thirty-three and sixty-four patients were evaluable in arm A and B, respectively. The overall response rate was 18% in arm A (95% CI: 7%-34%) and 40% in arm B (95% CI: 28%-52%). Median time to progression, median duration of response and survival were similar in both groups. Responders (CR + PR) survived statistically longer than non-responders only in arm B (20 vs. 10 months, P = 0.0016). All patients were evaluable for toxicity which was mild in both groups; gastrointestinal disturbances were the most common. There were no treatment-related deaths. Grade 3-4 toxicity was uncommon in both arms. CONCLUSIONS: The addition of CPT-11 to the hybrid LV5FU2 regimen provided a significant overall response rate (40%) with relatively mild toxicity. The overall response rate was 18% in patients treated with LV5FU2 alone in the calibration arm. Thus, considering other encouraging data from the literature, the CPT-11 + FU-LV combination therapy can be regarded as a new, very effective treatment option for first-line treatment of advanced colorectal cancer patients. PMID- 11038045 TI - Immunohistochemical determination of p53 protein does not predict clinical response in advanced colorectal cancer with low thymidylate synthase expression receiving a bolus 5-fluorouracil-leucovorin combination. AB - BACKGROUND: We assessed the hypothesis that a compromised p53 function could account for the non response of colon cancer patients with low thymidylate synthase (TS) expression receiving a bolus 5-fluorouracil (5-FU) leucovorin (LV) combination. PATIENTS AND METHODS: The study population consisted of 41 patients with unresectable metastatic colon cancer, homogeneously, treated with bolus 5-FU and LV. RESULTS: Twenty-seven patients (66%) showed high levels of TS expression. The difference in the proportion of objective responses between patients with low (CR + PR: 7 of 14, 50%) and high (CR + PR: 0 of 27) TS levels was statistically significant (P = 0.0001, chi-square test). p53 nuclear over-expression was found in 27 of 41 patients (66%). No differences were observed in p53 overexpression in patients with high (66%) or low (66%) TS expression. p53 status was not found to be associated with response even in patients with low TS expression. CONCLUSIONS: p53 status measured by immunohistochemistry does not seem to be useful to identify unresponsive patients with low TS expression. PMID- 11038047 TI - Syndrome of inappropriate antidiuretic hormone associated with chemotherapy induced tumour lysis in small-cell lung cancer: case report and literature review. AB - A patient with a small-cell lung cancer (SCLC) developed an asymptomatic hyponatremia, with all features of the syndrome of inappropriate antidiuretic hormone secretion (SIADH), two days after the start of his first cycle of chemotherapy with vindesine, ifosfamide and cisplatin. Progression of the tumour with an increase in paraneoplastic SIADH, or drug-induced causes of hyponatremia, could be ruled out by his further clinical course. The event was interpreted as a consequence of ADH release during the initial tumour cell lysis after effective chemotherapy. The occurrence of hyponatremia during the initial phase of chemotherapy for SCLC should be interpreted with caution. Although it is most commonly due to an increase in paraneoplastic ADH secretion reflecting ineffective therapy, it can also be due to release of ADH from malignant cells in the period of rapid tumour lysis, reflecting effective therapy. Based on this rare occurrence, a review of the aetiology, clinical findings, diagnosis, prognosis and treatment of SIADH in general is presented. PMID- 11038046 TI - Response to primary chemotherapy in breast cancer patients with tumors not expressing estrogen and progesterone receptors. AB - BACKGROUND: We recently demonstrated that in premenopausal patients with estrogen receptors (ER)-absent tumors, early initiation of systemic chemotherapy after primary surgery might improve outcome. These data indicate a different responsiveness to chemotherapy for tumors not expressing hormone receptors. To test this hypothesis we evaluated the responsiveness to preoperative chemotherapy in patients with ER and progesterone receptors (PgR)-absent tumors. PATIENTS AND METHODS: Patients with biopsy-proven T2-T3, N0-2 breast cancer treated at a single institution from January 1995 to August 1999 with preoperative chemotherapy were retrospectively evaluated. ER and PgR were determined immunohistochemically and classified for this purpose as absent (0% of the cells positive) or positive (> or = 1% of the cells). RESULTS: On 117 evaluable patients 72 had an objective response (61%). A significant difference in response was observed for patients with ER and PgR absent compared with those with ER and/or PgR-positive tumors (82% vs. 57%, P = 0.03 Fishers's exact test). Pathological complete remission rates were also significantly different in the two groups (23% vs. 7%, respectively; P = 0.04). CONCLUSIONS: The different degree of response according to hormone receptors expression supports the hypothesis that tumors not expressing both ER and PgR might represent a different clinical entity in terms of chemotherapy responsiveness. PMID- 11038049 TI - Can mitomycin C represent a valid partner for 5-fluorouracil in second-line chemotherapy of colorectal cancer? PMID- 11038048 TI - Tamoxifen-induced severe hypertriglyceridemia and pancreatitis. AB - Tamoxifen exhibits favorable effects on the lipid and lipoprotein profile since it decreases the total and LDL cholesterol levels as well as the Lp(a) levels. Additionally, a small increase in serum triglycerides is commonly found after tamoxifen administration. However, severe hypertriglyceridemia which can sometimes be associated with life-threatening complications is occasionally noticed. Herein, we describe a patient who developed tamoxifen-induced severe hypertriglyceridemia and pancreatitis. An analysis of the underlying pathogenetic mechanisms as well as a review of the relevant literature is also provided. PMID- 11038050 TI - Illumination increases the affinity of phosphoenolpyruvate carboxylase to bicarbonate in leaves of a C4 plant, Amaranthus hypochondriacus. AB - Illumination increased markedly the affinity to bicarbonate of phosphoenolpyruvate carboxylase (PEPC; EC 4.1.1.31) in leaves of Amaranthus hypochondriacus L., a C4 plant. When leaves were illuminated, the apparent Km for (HCO3-) of PEPC decreased by about 50% concurrent with a 2- to 5-fold increase in Vmax and 3- to 4-fold increase in Ki for malate. The inclusion of ethoxyzolamide, an inhibitor of carbonic anhydrase, during the assay had no effect on kinetic and regulatory properties of PEPC indicating that carbonic anhydrase was not involved during light-induced sensitization of PEPC to HCO3-. Pretreatment of leaf discs with cycloheximide (CHX), a cytosolic protein synthesis inhibitor, suppressed significantly the light-enhanced decrease in apparent Km (HCO3-). Further, in vitro phosphorylation of purified dark-form PEPC by protein kinase A (PKA) decreased the apparent Km (HCO3-) of the enzyme, in addition increasing Ki (malate) as expected. Such changes, due to in vitro phosphorylation of purified PEPC by PKA, occurred only with wild-type PEPC, but not in the mutant form of maize (S15D) which is already a mimic of the phosphorylated enzyme. These results suggest that phosphorylation of the enzyme is important during the sensitization of PEPC to HCO3- by illumination in C4 leaves. Since illumination is expected to increase the cytosolic pH and the availability of dissolved HCO3- in mesophyll cells, the sensitization by light of PEPC to HCO3- could be physiologically quite significant. PMID- 11038051 TI - Genetic engineering on shikonin biosynthesis: expression of the bacterial ubiA gene in Lithospermum erythrorhizon. AB - The naphthoquinone pigment shikonin from Lithospermum erythrorhizon Sieb. et Zucc. (Boraginaceae) was the first plant secondary metabolite produced in industrial scale from plant cell cultures. We have now manipulated the biosynthetic pathway leading to shikonin in L. erythrorhizon by introduction of the bacterial gene ubiA. This gene of Escherichia coli encodes 4-hydroxybenzoate 3-polyprenyltransferase, a membrane-bound enzyme that catalyzes a key step in ubiquinone biosynthesis. Using geranyl diphosphate (GPP) as substrate, it is able to catalyze the formation of 3-geranyl-4-hydroxybenzoate (GBA), a principal step of shikonin biosynthesis. The prokaryotic ubiA gene was fused to two signal sequences for targeting of the resulting peptide to the endoplasmic reticulum (ER). Constructs with different constitutive promoters were introduced into L. erythrorhizon using Agrobacterium rhizogenes-mediated transformation. In the resulting hairy root lines, high UbiA enzyme activities could be observed, reaching 133 pkat mg(-1). Expression of ubiA resulted in an accumulation of GBA in an amount exceeding that of the control culture by a factor of 50. However, the ubiA-transformed lines showed only a marginal (average 22%) increase of shikonin production in comparison to the control lines, and there was no significant correlation of UbiA enzyme activity and shikonin accumulation. This suggests that overexpression of ubiA alone is not sufficient to increase shikonin formation, and that further enzymes are involved in the regulation of this pathway. PMID- 11038052 TI - Isolation of a novel 190 kDa protein from tobacco BY-2 cells: possible involvement in the interaction between actin filaments and microtubules. AB - Interaction between actin filaments (AFs) and microtubules (MTs) has been reported in various plant cells, and the presence of a factor(s) connecting these two cytoskeletal networks has been suggested, but its molecular entity has not been elucidated yet. We obtained a fraction containing MT-binding polypeptides, which induced bundling of AFs and of MTs. A 190 kDa polypeptide which associated with AFs was selectively isolated from the fraction. This polypeptide was thought to have an ability to bind to both AFs and MTs. We raised a monoclonal antibody against the 190 kDa polypeptide. Immunostaining demonstrated the association of the 190 kDa polypeptide with AF bundles and with MT bundles formed in vitro. Immunocytochemical studies throughout the cell cycle revealed that the 190 kDa polypeptide was localized in the nucleus before nuclear envelope breakdown, and in the spindle and the phragmoplast during cell division. After the re-formation of the nuclear envelope, the 190 kDa polypeptide was sequestered to the daughter nuclei. Using the antibody, we succeeded in cloning a cDNA encoding the 190 kDa polypeptide. PMID- 11038053 TI - Regulation of elongation growth by gibberellin in root segments of Lemna minor. AB - Hormonal control of elongation growth was analyzed in segments excised from the elongation zone of Lemna roots. Exogenous GA3 did not promote the segment elongation but rather inhibited it. Uniconazole-P, a gibberellin biosynthesis inhibitor, significantly inhibited the segment elongation, and the inhibitory effect was completely nullified by GA3. In the epidermis, cell elongation was inhibited, but lateral cell expansion was not affected by uniconazole-P. Orientation of cortical microtubules of epidermal cells was disturbed by treatment with uniconazole-P for 12 h, and the disorganization of cortical microtubules was ameliorated by GA3. These findings suggested that disorganization of cortical microtubules induced inhibition of elongation growth of root. However, stabilization of cortical microtubules by taxol, a microtubule stabilizing agent, did not affect the inhibition of segment elongation by uniconazole-P. These results suggested that endogenous gibberellin controls the elongation growth of root by regulating cell elongation. PMID- 11038054 TI - Characterization and expression of monosaccharide transporters (osMSTs) in rice. AB - This study deals with the cloning and characterization of monosaccharide transporter cDNAs in rice. OsMST1-3 (Oryza sativa monosaccharide transporters 1 3) have two sets of putative six transmembrane domains separated by a central long hydrophilic region. Heterologous expression of OsMST3 in the yeast Saccharomyces cerevisiae indicated that OsMST3 has transport activity for some monosaccharides in an energy-dependent H+ co-transport manner. Northern blot and in situ hybridization analyses showed that OsMST3 mRNA is detectable in leaf blades, leaf sheaths, calli and roots, especially the xylem as well as in sclerenchyma cells in the root. These results suggested that OsMST3 is involved in the accumulation of monosaccharides required for cell wall synthesis at the stage of cell thickening. PMID- 11038055 TI - Highly substituted glucuronoarabinoxylans (hsGAXs) and low-branched xylans show a distinct localization pattern in the tissues of Zea mays L. AB - Polyclonal antibodies which recognized highly substituted glucuronoarabinoxylans (hsGAXs) and low-branched xylans and did not cross-react with each other, were raised in order to examine localization of these epitopes in internodes of maize. Immunofluorescent labeling revealed different pattern between two succeeding developmental stages. The hsGAX epitope was localized evenly in primary walls in all tissue types, and strongly in unlignified secondary walls in phloem. However, lignified secondary walls in protoxylem, parenchyma and a part of fibers were faintly labeled with this epitope. Moreover, the epitope showed limited binding in lignified parenchyma and fiber walls at ultrastructural level. Low-branched xylan epitope was localized evenly throughout lignified walls in all tissue types. This epitope was also localized only in lignified walls of other organs such as leaf, root apex and dark-grown mesocotyl. Low-branched xylans are significantly related to lignification. Localization of hsGAX epitope in their organs was similar to that in internodes. The hsGAX epitope was distributed both in unlignified walls of all tissues and in lignified walls of parenchyma and annular thickening of protoxylem. We propose that hsGAX has separate functions in lignified and unlignified tissues. In conclusion, at tissue level, hsGAX is localized mainly in unlignified walls, and low-branched xylans in lignified walls. PMID- 11038056 TI - Are isocitrate lyase and phosphoenolpyruvate carboxykinase involved in gluconeogenesis during senescence of barley leaves and cucumber cotyledons? AB - The aim of this study was to investigate whether gluconeogenesis catalysed by phosphoenolpyruvate carboxykinase (PEPCK) occurs during leaf senescence. This was addressed by determining changes in the abundance and intercellular location of enzymes necessary for gluconeogenesis during the senescence of barley leaves and cucumber cotyledons. PEPCK was never present in barley leaves, despite the presence of large amounts of isocitrate lyase (ICL), a key enzyme of the glyoxylate cycle, and of its product, glyoxylate. Although PEPCK was present in non-senescent cucumber cotyledons, its abundance declined during senescence. Throughout senescence, PEPCK was only present in the trichomes and vasculature, whereas ICL was located in mesophyll cells. Pyruvate,Pi dikinase (PPDK) which, in concert with NAD(P)-malic enzyme, is also capable of catalysing gluconeogenesis, was present in non-senescent barley leaves and cucumber cotyledons, but in both plants its abundance decreased greatly during senescence. The abundance of ICL was greatly reduced in senescing detached barley leaves by either illumination or by co-incubation with sucrose, and greatly increased in darkened attached barley leaves. These results argue against the large-scale occurrence of gluconeogenesis during senescence catalysed either by PEPCK or PPDK. In cucumber cotyledons, PEPCK may play a role in metabolic processes linked to the export of amino acids, a role in which phosphoenolpyruvate carboxylase may also be involved. The amount of ICL was increased by starvation and during senescence may function in the conversion of lipids to organic acids, which are then utilised in the mobilisation of amino acids from leaf protein. PMID- 11038057 TI - Possible involvement of 65 kda MAP in elongation growth of azuki bean epicotyls. AB - Although regulation of the dynamics of plant microtubules (MTs) by microtubule associated proteins (MAPs) has been suggested, the mechanism has not yet been elucidated. As one candidate, a MAP composed of a 65 kDa polypeptide (65 kDa MAP) has been isolated from tobacco cultured cells [Jiang and Sonobe (1993), J. Cell Sci 105: 8911. To investigate the physiological role of the 65 kDa MAP in situ, we analyzed the changes in content and colocalization of this MAP with cortical MTs in relation to elongation growth, using azuki bean epicotyls (Vigna angularis Ohwi et Ohashi). All apical, intermediate, and basal segments prepared from 6 d seedlings showed high growth activity. In 12 d seedlings, growth activity of intermediate and basal segments was low, although that of apical segments was high. The relationship between the growth activity and the orientation of cortical MTs in the epidermal cells was analyzed. Cells could be classified into four types with respect to orientation of cortical MTs: transverse (T), oblique (O), longitudinal (L) to the vertical axis of cells, and random (R). In rapidly growing segments, three types of cells, T, O, L, were observed at similar ratios. In such segments, significant amounts of the 65 kDa MAP were expressed, and it colocalized well with cortical MTs. In segments showing low growth activity, most of the cells showed oblique and longitudinal orientation of cortical MTs. In such segments, the content of the 65 kDa MAP was low. These results suggested involvement of this 65 kDa MAP in regulation of the elongation growth of this epicotyl. PMID- 11038058 TI - Ser162-Dependent inactivation of overproduced sucrose-phosphate synthase protein of maize leaf in transgenic rice plants. AB - To investigate the role of Ser162 in phosphorylation-dependent regulation of maize sucrose-phosphate synthase (SPS) activities in rice, transgenic rice plants expressing wild-type or mutagenized maize SPS were produced. Our results indicate that Ser162 was responsible for overproduction-induced inactivation of SPS protein and for light/ dark modulation of this protein in vivo. PMID- 11038059 TI - Induction of chalcone synthase expression in white spruce by wounding and jasmonate. AB - The phenylpropanoid pathway has important functions in angiospermous plants following exposure to environmental stresses, such as wounding and pathogen attack, that lead to production of compounds including lignin, flavonoids and phytoalexins. Chalcone synthase (CHS) is a key enzyme in this pathway, catalyzing the first step in flavonoid biosynthesis, whose expression can be induced in response to environmental stress. To explore the response of conifers to environmental stress, expression of spruce CHS and its inducibility were investigated. A partial spruce CHS cDNA clone was isolated using PCR. Examination of the expression patterns of the CHS gene family in white spruce revealed accumulation of CHS mRNA in needle tissue following mechanical wounding, or application of signal molecules like jasmonic acid or methyl jasmonate. Repeated mechanical wounding or jasmonate applications had an enhancing effect on transcript accumulation in needles. A systemic accumulation of CHS mRNAs following wounding was also observed. Conifers thus appear to possess a general wound response similar to that found for angiosperms, which includes CHS induction as well as its inducibility by jasmonic acid and airborne methyl jasmonate. PMID- 11038060 TI - A novel glycolate oxidase requiring flavin mononucleotide as the cofactor in the prasinophycean alga Mesostigma viride. AB - A new type of glycolate-oxidizing enzyme was found in the prasinophycean alga Mesostigma viride. This enzyme was characterized as the glycolate oxidase that strongly requires FMN, thus distinguishing it from most other Prasinophytes that possess glycolate dehydrogenase instead. PMID- 11038061 TI - Comparison of cancellous bone-derived cell proliferation in autologous human and fetal bovine serum. AB - Conventionally, culture medium is supplemented with fetal bovine serum (FBS): such serum presents potential risks of foreign protein contamination and transmission of viral or prion-related disease if used in culture of cells intended for human reimplantation. As it has been suggested that a composite of cultured human cancellous bone-derived cells and a bone graft substitute may present a solution to the well-recognized complications and limited availability associated with harvest of fresh bone graft, this study aimed to compare the proliferative response of human cancellous bone-derived cells supplemented with FBS or autologous human serum (AHS) to determine whether AHS is a practical alternative. Explant cultures were established using greater trochanter trabecular bone from 10 consenting patients (aged 57-84) undergoing total hip arthroplasty. At the same time, serum was harvested. The cells were characterized by alkaline phosphatase expression and by in vitro mineralization in enhanced medium. At confluence, cells were aliquoted into multiwell plates and grown for 9 days in medium supplemented with 5%, 10%, 15%, or 20% AHS or 10% FBS. Proliferative response was determined by a crystal violet dye binding assay. There was no significant difference between proliferation in 5% AHS and 10% FBS. However, 10%, 15%, and 20% AHS all produced significantly greater proliferation than 10% FBS. The proliferative response was dose related. FBS is said to be rich in growth and attachment factors, which is why it is widely used in tissue culture. These results suggest that species specificity, even when using adult serum, outweighs these advantages. It should therefore be considered as a prerequisite for any program involving reimplantation of cultured human cells. Clinical trials of cultured human cancellous bone-derived cells have now begun. PMID- 11038062 TI - Reduced NO production improves early canine islet xenograft function: a role for nitric oxide in islet xenograft primary nonfunction. AB - Isolated canine islets transplanted to hyperglycemic rats fail to restore euglycemia in almost all cases, although the grafted islet tissue appears to be morphologically intact for up to 48 h following transplantation. Cytokines typically produced in the xenograft environment (e.g., IL-1 and TNF) inhibit insulin biosynthesis and secretion from isolated pancreatic islets, and are associated with the production of nitric oxide (NO). To further define the relationship between NO production and islet xenotransplantation, the inhibition of NO in a splenocyte/islet coculture system, and the in vivo effect of this inhibition on canine islet xenotransplantation, was investigated. Splenocytes (SPLC) from Lewis rats were cocultured with canine islets (freshly isolated or cultured 7 days), supernatant removed, and NO concentration (NO2) determined by optical density (Griess reaction, 550 nm, expressed as nmol nitrite/10(6) cells/18 h). Lipopolysaccharide (LPS) was used as a positive control of SPLC production of NO. Stimulation by LPS resulted in maximal NO production (2.20 +/- 0.16 nmol/10(6) cells/18 h, p < 0.001 compared to baseline values of 0.73 +/- 0.04 nmol/10(6) cells/18 h). In the presence of NO inhibitors (NMA, polymyxin B, hydrocortisone, aminoguanidine, DMSO), nitrite levels did not significantly rise above unstimulated values. Freshly isolated canine islets did stimulate NO production (1.26 +/- 0.12 nmol/10(6) cells/18 h, p < 0.001). In contrast, cultured canine islets did not stimulate NO production (0.84 +/- 0.09 nmol/10(6) cells/18 h). Transplantation of freshly isolated canine islets to STZ-diabetic recipient Lewis rats resulted in amelioration of hyperglycemia in only 50% (n = 6) of recipients 12 h posttransplant, with a return to hyperglycemia at all subsequent time points. Transplantation of 7-day cultured canine islets resulted in amelioration of hyperglycemia in 88% of recipients 12 h posttransplant and 63% of recipients 24 h posttransplant [p = 0.028, mean survival time (MST) = 1.0 days, n = 8]. Transplantation of canine islet xenografts with aminoguanidine therapy (BID, n = 11) resulted in amelioration of hyperglycemia in 100% of recipients at 12 h posttransplant, decreasing to 82% by 24 h following transplantation (p = 0.002, MST = 0.9 days). These results demonstrate that freshly isolated canine islets are potent stimulators of NO production by rat SPLC in vitro, and that culture of canine islets, or addition of NO inhibitors, abrogates stimulated NO production. These results also demonstrate a statistically significant improvement (p < 0.001) in early function of canine islet xenografts following 7 days of islet culture prior to transplant, and following recipient treatment with aminoguanidine. These studies suggest that the production of NO in the microenvironment of the graft site may adversely affect engraftment and function of canine islets, and suggest that the abrogation of islet-stimulated NO production may improve engraftment following islet xenotransplantation. PMID- 11038063 TI - The efficiency of muscle-derived cell-mediated bone formation. AB - The development of new clinically applicable methods for the delivery of bone morphogenic protein (BMP) is an area of intensive research. Cell-mediated gene therapy approaches are being explored as a potential delivery vehicle. Primary muscle-derived cells isolated from an adult mouse were transduced with an adenoviral-BMP-2 construct. These cells were injected into the triceps surae of severe combined immune deficient (SCID) mice where they induced heterotopic bone formation. BMP-2 expression by these muscle-derived cell constructs was measured in vitro to estimate in vivo BMP-2 delivery. In vitro expression of BMP-2 by 3 x l0(5) muscle-derived cells was 87.89 ng/72 h. These results suggest that the efficiency of muscle cell-based gene delivery of BMP-2 exceeds the direct delivery of recombinant BMP-2 protein. PMID- 11038064 TI - Evaluation of an intrathecal immune response in amyotrophic lateral sclerosis patients implanted with encapsulated genetically engineered xenogeneic cells. AB - A phase I/II clinical trial has been performed in 12 amyotrophic lateral sclerosis (ALS) patients to evaluate the safety and tolerability of intrathecal implants of encapsulated genetically engineered baby hamster kidney (BHK) cells releasing human ciliary neurotrophic factor (CNTF). These patients have been assessed for a possible intrathecal or systemic immune response against the implanted xenogeneic cells. Hundreds of pg CNTF/ml could be detected for several weeks in the cerebrospinal fluid (CSF) of 9 out of 12 patients, in 2 patients up to 20 weeks after capsule implantation. Slightly elevated leukocyte counts were observed in 6 patients. Clear evidence for a delayed humoral immune response was found in the CSF of only 3 patients out of 12 (patients #4, #6, and #10). Characterization of the antigen(s) recognized by the antibodies present in these CSF samples allowed to identify bovine fetuin as the main antigenic component. The defined medium used for maintaining the capsules in vitro before implantation contains bovine fetuin. Fetuin may therefore still be adsorbed to the surface of the cells and/or the polymer membrane, or be present in the medium surrounding the encapsulated cells at the time of implantation. Because of the insufficient availability of CSF samples, as well as the relatively poor sensitivity of the assays used, a weak humoral immune response against components of the implanted cells themselves cannot be excluded. However, the present study demonstrates that encapsulated xenogeneic cells implanted intrathecally can survive for up to 20 weeks in the absence of immunosuppression and that neither CNTF nor the presence of antibodies against bovine fetuin elicit any adverse side effects in the implanted patients. PMID- 11038065 TI - Myoblast transfer therapy in the new millennium. PMID- 11038066 TI - Why do cultured transplanted myoblasts die in vivo? DNA quantification shows enhanced survival of donor male myoblasts in host mice depleted of CD4+ and CD8+ cells or Nk1.1+ cells. AB - Overcoming the massive and rapid death of injected donor myoblasts is the primary hurdle for successful myoblast transfer therapy (MTT), designed as a treatment for the lethal childhood myopathy Duchenne muscular dystrophy. The injection of male myoblasts into female host mice and quantification of surviving male DNA using the Y-chromosome-specific (Y1) probe allows the speed and extent of death of donor myoblasts to be determined. Cultured normal C57BL/10Sn male donor myoblasts were injected into untreated normal C57BL/10Sn and dystrophic mdx female host mice and analyzed by slot blots using a 32P-labeled Y1 probe. The amount of male DNA from donor myoblasts showed a remarkable decrease within minutes and by 1 h represented only about 10-18% of the 2.5 x 10(5) cells originally injected (designated 100%). This declined further over 1 week to approximately 1-4%. The host environment (normal or dystrophic) as well as the extent of passaging in tissue culture (early "P3" or late "P15-20" passage) made no difference to this result. Modulation of the host response by CD4+/CD8+ depleting antibodies administered prior to injection of the cultured myoblasts dramatically enhanced donor myoblast survival in dystrophic mdx hosts (15-fold relative to untreated hosts after 1 week). NK1.1 depletion also dramatically enhanced donor myoblast survival in dystrophic mdx hosts (21-fold after 1 week) compared to untreated hosts. These results provide a strategic approach to enhance donor myoblast survival in clinical trials of MTT. PMID- 11038067 TI - The influence of muscle fiber type in myoblast-mediated gene transfer to skeletal muscles. AB - Myoblast transplantation has been hindered by immune rejection problems, as well as the poor survival and spread of transplanted cells. Our recent study has shown that the poor survival of the injected cells can be totally overcome by the use of specific populations of muscle-derived cells. In the present study, we have investigated whether a relationship exists between the fate of transplanted cells and the muscle fiber types. Four kinds of myogenic cells [primary myoblasts at a high purity (PMb), myoblasts isolated from fast single fibers (FMb), mdx (MCL), and MtMd-1 cell lines] were infected with an adenoviral vector carrying a LacZ reporter gene and injected into mdx hindlimb muscle. The LacZ transduced myofibers formed by the fusion of the injected myoblasts at 2-10 days postinjection were colocalized with MyHC stainings. The PMb cells, which expressed both slow and fast MyHCs in vitro, displayed the same phenotypes when injected into the m. soleus and m. gastrocnemius (white) muscles, which contained 70% and 0% of slow myofibers, respectively, and showed a high degree of fusion with host muscle fibers. In contrast, the FMb cells only expressed fast MyHCs in vitro and fused exclusively with each other or with host fast muscle fibers when injected in the m. gastrocnemius. Injected MCL and MtMd-1 fused predominantly with each other and displayed a similar expression of MyHCs to those they expressed in vitro. Just a few host myofibers were found to express the reporter gene product following implantation of both cell lines, indicating that these myogenic cell lines display an intrinsic potential to fuse together rather than with host myofibers. Based on the data, we concluded that 1) the essential key to survival is the ability of the donor cells to fuse with the host myofibers, and 2) the most successful combination is achieved between donor primary muscle cells that express both fast and slow MyHC and a host muscle type that facilitates fusion. PMID- 11038068 TI - A factor implicated in the myogenic conversion of nonmuscle cells derived from the mouse dermis. AB - Using the mdx mouse model for human Duchenne muscular dystrophy we have shown that a cell population residing in the dermis of C57B1/10ScSn mouse skin is capable of converting to a myogenic lineage when implanted into the mdx muscle environment. It was important to determine the characteristics of the converting cell. A previous in vitro study indicated that 10% of cells underwent conversion but only when the cells were grown in medium previously harvested from a myogenic culture. In the present study we cloned cells derived from the dermis to identify the converting cells. Clones grown in normal growth medium showed no conversion, but when grown in medium conditioned by muscle cells around 40% conversion was achieved in several individual clones. We investigated whether the protein beta galactoside binding protein (betaGBP), which is secreted by myoblasts and acts as a cell growth regulator of fibroblasts. could be a candidate factor responsible for conversion. Medium harvested from COS-1 cells infected with a construct containing betaGBP has been used for this investigation. Growth of dermal fibroblasts in medium enriched with this factor showed a high rate of conversion to cells expressing muscle-specific factors. PMID- 11038069 TI - Thymic myoid cells as a source of cells for myoblast transfer. AB - Transplantation of disaggregated myoblasts from normal donor to the muscles of a diseased host, or reimplantation of genetically modified host myoblasts, has been suggested as a possible route to therapy for inherited myopathies such as Duchenne muscular dystrophy, or to supply missing proteins that are required systemically in diseases such as hemophilia. With two exceptions, studies of myoblast transfer in the mouse have involved transplantation of donor myoblasts isolated from adult or neonatal skeletal muscle satellite cells. In this study we present evidence that thymic myoid cells are capable of participating in the regeneration of postnatal skeletal muscle, resulting in the expression of donor derived proteins such as dystrophin and retrovirally encoded proteins such as beta-galactosidase within host muscles. This leads us to conclude that thymic myoid cells may provide an alternative to myoblasts derived from skeletal muscle as a source of myogenic cells for myoblast transfer. PMID- 11038070 TI - Intramuscular migration of myoblasts transplanted after muscle pretreatment with metalloproteinases. AB - The effect of pretreatments of host muscles with metalloproteinases (MMPs) or with notexin on the migration of transplanted myoblasts was investigated. Transgenic TnILacZ mice in which the beta-galactosidase gene is under the control of a quail fast skeletal troponin I gene promoter were used as donors. A polyethylene microtube with four perforations was inserted in the tibialis anterior (TA) of CD1 mice. Both pretreatment substances and cells were slowly injected through that microtube. Muscles were pretreated 2 days before myoblast injection either with a mixture of collagenase, matrilysin, and notexin or with only collagenase and matrilysin or only notexin. As control for our experiments, TnILacZ and C2C12 myoblasts were also injected in TA muscles not pretreated. Comparison of short and long-term myoblast radial migration was performed using a dye (PKH26) and X-gal staining, respectively. The recipient mice were immunosuppressed with FK506. Two days after myoblast transplantation, the cell movement in muscles pretreated with collagenase, matrilysin, and notexin was slightly greater than in muscles pretreated only with collagenase and matrilysin but was about twice that observed in muscles treated with notexin alone. Almost no radial migration of TnILacZ myoblasts was observed in untreated muscles. The C2C12 myoblasts showed a four-to fivefold higher migration capacity than TnILacZ myoblasts. At 15 days after TnILacZ myoblast transplantation, the farthest positive beta-gal muscle fibers show a two- to threefold extension of the initial migration observed at 2 days, demonstrating the ability of myoblasts to continue the migration following all pretreatments and even in the untreated muscles. In addition, more muscle fibers expressed the beta-gal reporter gene in muscles pretreated only with MMPs. Our results clearly demonstrate that muscle pretreatments with MMPs increase myoblast migration and fusion with host muscle fibers after transplantation and that the C2C12 cell line producing MMPs has a higher migratory capacity. PMID- 11038071 TI - Deflazacort increases laminin expression and myogenic repair, and induces early persistent functional gain in mdx mouse muscular dystrophy. AB - Deflazacort slows the progress of Duchenne muscular dystrophy (DMD) with fewer side effects than prednisone. In mdx mice, deflazacort treatment augments repair and growth of new muscle fibers. We tested the hypothesis that deflazacort improves muscle function and promotes repair by increasing myogenic cell proliferation and fiber differentiation. mdx mice (3.5 weeks old) were treated with deflazacort (1.2 mg/kg) or vehicle for 4 weeks. Forelimb grip strength was measured. After 4 weeks, the right tibialis anterior muscle (TA) was crush injured to induce synchronous regeneration. DNA was labeled using different markers 24 and 2 h before collecting tissues 4 days after injury. The expression of creatine kinase (CK) isoforms, laminin-2 (merosin) mRNA and protein, and proliferation by myogenic cells were measured and satellite cells were identified by immunolocalization of c-met receptor. Peak grip strength increased 15% within 10 days of treatment, and was maintained up to 6 weeks after the end of treatment in a second experiment. Expression of CK MM in the regenerating TA rose from 46% to 55% of total CK activity after deflazacort treatment. Satellite cells were more numerous and appeared earlier on new fibers, in concert with a threefold increase in proliferation by myogenin+ (but not MyoD+) myoblasts. alpha2-Laminin mRNA expression and protein increased 1.3-5.5-fold relative to MM CK in regenerating and dystrophic TA, respectively. These studies support the hypothesis that deflazacort promotes functional gains, myogenic differentiation, myoblast fusion, and laminin expression in regenerating dystrophic muscle. The potential to augment precursor specification, strength, and possible membrane stability may be useful in directing long-term benefits for DMD patients and short-term amplification of precursors prior to myoblast transfer. PMID- 11038072 TI - Duty, trust, and the training of residents. PMID- 11038073 TI - Novel strategies for immunomodulation after trauma: revisiting hypertonic saline as a resuscitation strategy for hemorrhagic shock. PMID- 11038074 TI - Advanced or basic life support for trauma: meta-analysis and critical review of the literature. AB - BACKGROUND: The question of whether to use advanced life support (ALS) or basic life support (BLS) for trauma patients in the prehospital setting has been much debated and still lacks a clear answer. The purpose of this study was to conduct a comprehensive critical review of the literature regarding this controversy METHODS: A total of 174 articles on prehospital ALS or BLS for trauma were reviewed. Fifteen of these studies were found to involve mortality statistics for both ALS- and BLS-treated patients. Odds ratios were calculated for survival in ALS versus BLS and summarized across studies on the basis of multivariate scoring systems that incorporated both design and methodological assessment. Overall odds ratios for all studies were calculated on the basis of both raw data from the papers, and weighted odds ratios were calculated from the scoring systems. RESULTS: Six studies were scored as being methodologically average (5 favoring BLS and 1 favoring ALS), two were scored as good (1 favoring BLS and 1 favoring ALS), seven as excellent (6 favoring BLS and 1 favoring ALS). Ten studies had an average study design score (6 favoring BLS and 4 favoring ALS) and seven had a good study design score (6 favoring BLS and 1 favoring ALS). Weighted odds ratio for dying was 2.59 for patients receiving ALS compared with those receiving BLS. The crude odds ratio was 2.92. CONCLUSION: The aggregated data in the literature have failed to demonstrate a benefit for on-site ALS provided to trauma patients and support the scoop and run approach. PMID- 11038075 TI - Missed injuries in patients with multiple trauma. AB - BACKGROUND: Understanding the etiology of missed injuries is essential in minimizing its occurrence. A retrospective review was conducted to identify the incidence, contributing factors, and clinical outcomes of missed injuries. METHODS: All trauma patients assessed by St Michael's Hospital trauma service from April 1, 1995, to July 31, 1997, were included in the study. Demographic and medical data were compared and statistically analyzed in two patient groups to identify factors associated with missed injuries. RESULTS: Forty six of 567 patients (8.1%) had missed injuries. Patients with missed injuries had higher mean Injury Severity Scores and longer stays in the hospital and intensive care unit compared with patients without missed injuries (p < 0.05). Patients with missed injuries were more likely to have lower Glasgow Coma Scale scores and to have required pharmacologic paralysis (p < 0.05). Of the factors contributing to missed injuries, 56.3% were potentially avoidable and 43.8% were unavoidable. Seven patients with missed injuries had clinically significant outcomes, including one patient death. Of the seven clinically significant missed injuries, five were attributable to potentially avoidable factors. CONCLUSION: Patients with missed injuries tend to be more severely injured with initial neurologic compromise. The majority of missed injuries are potentially avoidable with repeat clinical assessments and a high index of suspicion. PMID- 11038076 TI - Effects of other intra-abdominal injuries on the diagnosis, management, and outcome of small bowel trauma. AB - BACKGROUND: Difficulty with and delays in diagnosis are possible causes of increased morbidity and mortality in small bowel injuries. We assessed whether multiple intra-abdominal injuries led to earlier laparotomy and whether this resulted in improved outcome. METHODS: Patients with small bowel injuries between January 1993 and December 1997 from the trauma database at St. Michael's Hospital in Toronto were assessed after dividing them into those with isolated small bowel injury ("isolated") and those with small bowel injuries in association with other intra-abdominal injuries ("nonisolated"). Parameters compared were age, gender, length of stay, mortality, intra-abdominal complications, mechanism of injury, diagnostic time, and how the diagnosis was made. RESULTS: Of 1,207 patients, 244 sustained abdominal injuries, and 83 had small bowel injuries (30 patients in the isolated group and 53 in the nonisolated group). Groups were similar with respect to age and gender, yet differed significantly with respect to mechanism and mean Injury Severity Scores (isolated, 18 +/- 8 vs. nonisolated, 30 +/- 15). Outcome differed between groups, as mortality (isolated, 0 of 30 vs. nonisolated, 4 of 53 deaths), length of stay (isolated, 13 +/- 2 vs. nonisolated, 22 +/- 3 days), and patients with intra-abdominal complications (isolated, 5 of 30 vs. nonisolated, 14 of 53 patients) were significantly higher in the nonisolated group. Time to diagnosis was significantly less in the nonisolated group. Decision for laparotomy and diagnosis of small bowel injuries were based more on physical findings in the nonisolated group and on computed tomography in the isolated group. CONCLUSION: The presence of associated intra-abdominal injuries significantly affects presentation and outcome of patients with small bowel injuries and the selection of diagnostic modalities. PMID- 11038077 TI - Recombinant humanized monoclonal antibody against CD18 (rhuMAb CD18) in traumatic hemorrhagic shock: results of a phase II clinical trial. Traumatic Shock Group. AB - BACKGROUND: Activated neutrophils have been shown to play a pivotal role in resuscitation injury after traumatic hemorrhagic shock. Blocking the adhesion of neutrophils with a recombinant humanized monoclonal antibody against CD18 (rhuMAb CD18) may reduce resuscitation injury but increase the risk of infection. This was a dose-finding phase II study to determine safety, pharmacokinetics, pharmacodynamics, and clinical outcome parameters for additional studies. METHODS: This was a prospective, placebo-controlled, randomized (3:1), double blind phase II trial enrolling 116 blunt and penetrating trauma patients from 14 trauma centers over a 9-month period. Patients with hypotension (blood pressure < or =90 mm Hg) from hemorrhagic shock were given a single intravenous dose of rhuMAb CD18 or placebo. The three doses tested were 0.5, 1, and 2 mg/kg. The drug was administered within 4 hours of the hypotensive episode and no later than 6 hours from time of injury. Exclusion criteria included head injury resulting in Glasgow Coma Scale score less than 8 or a history of cardiopulmonary resuscitation in the trauma center. An independent Drug Safety and Monitoring Review Board periodically reviewed unblinded data for safety issues and to give approval for dose escalation. RESULTS: Minor and major infection rates in rhuMAb CD18 groups were comparable to placebo. There was no evidence of antibody formation against rhuMAb CD18. Linear PK was observed within the dose range studied. Duration of neutrophil binding was dose-dependent, with 2 mg/kg resulting in greater than 90% neutrophil CD18 receptor saturation for approximately 48 hours. The mortality was 6.7% (2 of 30) in the placebo group, 4.8% (1 of 21) in the 0.5-mg/kg group, 8.5% (4 of 47) in the 1-mg/kg group, and 0% (0 of 18) in the 2-mg/kg group. The study was not powered for efficacy, and none of the efficacy variables demonstrated statistical significance. Favorable trends were seen in the 2-mg/kg group as compared with placebo in median intensive care unit length of stay (5 vs. 9 days) and median time on ventilator (34 vs. 72 hours). CONCLUSIONS: A single 2-mg/kg dose of rhuMAb CD18 maintains greater than 90% saturation of neutrophil CD18 receptors for approximately 48 hours in patients with traumatic hemorrhagic shock undergoing resuscitation. There was no trend toward increased infection. A larger trial is needed to demonstrate the clinical efficacy of rhuMAb CD18, perhaps using more reliable endpoints. PMID- 11038078 TI - Abdominal perfusion pressure: a superior parameter in the assessment of intra abdominal hypertension. AB - OBJECTIVE: To assess the clinical utility of abdominal perfusion pressure (mean arterial pressure minus intra-abdominal pressure) as both a resuscitative endpoint and predictor of survival in patients with intra-abdominal hypertension. METHODS: 144 surgical patients treated for intra-abdominal hypertension between May 1997 and June 1999 were retrospectively reviewed. Multivariate logistic regression and receiver operating characteristic curve analysis of common physiologic variables and resuscitation endpoints were performed to determine the decision thresholds for each variable that predict patient survival. RESULTS: Abdominal perfusion pressure was statistically superior to both mean arterial pressure and intravesicular pressure in predicting patient survival from intra abdominal hypertension and abdominal compartment syndrome. Multiple regression analysis demonstrated that abdominal perfusion pressure was also superior to other common resuscitation endpoints, including arterial pH, base deficit, arterial lactate, and hourly urinary output. CONCLUSION: Abdominal perfusion pressure appears to be a clinically useful resuscitation endpoint and predictor of patient survival during treatment for intra-abdominal hypertension and abdominal compartment syndrome. PMID- 11038079 TI - Ostomy as a risk factor for posttraumatic infection in penetrating colonic injuries: univariate and multivariate analyses. AB - BACKGROUND: Primary repair for penetrating colonic injury is an acceptable practice in uncomplicated injuries, but it is still viewed with trepidation in high risk patients. METHODS: The records of 350 patients evaluated at an urban Level I trauma center for penetrating colonic injuries over an 8-year period (1989-1997) were reviewed. These included 33 stab and 317 gunshot wounds. Thirty nine patients died within 48 hours. Of the remaining 311 patients, 78 (25%) developed 152 infections. These infections were classified as traumatic or nosocomial in nature. Traumatic infections (46%) included abdominal abscesses or peritonitis (28), wound infections (30), missile tract infections (8), and fistulas (4), whereas nosocomial infections (54%) included pneumonia (25), bacteremia (25), urinary tract infections (17), miscellaneous (8), empyema (4), and sinusitis (3). Significance for analyses was set at p < 0.05. RESULTS: Univariate analysis was performed to identify risk factors for the development of infections. The five most significant risk factors, using all infections as an outcome, were as follows: penetrating abdominal trauma index (PATI) greater than 30, presence of an ostomy, multiple transfusions, Injury Severity Score (ISS) of 16 or greater, and Revised Trauma Score less than 7.8. All were highly significant (p < 0.0001). Multivariate analysis with all infections as an outcome revealed that four of the five risk factors had independent effects, with the following significance: PATI greater than 30, ISS of 16 or greater, ostomy, and multiple transfusions. Multivariate analysis for traumatic infections revealed only two of the above to be independent risk factors: presence of an ostomy (p = 0.004) and a PATI greater than 30 (p = 0.039), both of which can be considered local factors. Conversely, multivariate analysis of nosocomial infections revealed independent risk for the two other factors, both of which can be considered systemic factors: multiple transfusions (p = 0.011) and ISS of 16 or greater (p = 0.026). CONCLUSION: Although most of the above factors are beyond the control of the trauma surgeon, the creation of an ostomy is a clinical decision. The creation of an ostomy in high-risk patients does not protect them from septic complications and, indeed, may independently contribute to local abdominal infections. PMID- 11038080 TI - Incidence and susceptibility of pathogenic bacteria vary between intensive care units within a single hospital: implications for empiric antibiotic strategies. AB - BACKGROUND: The purpose of this study was to determine whether the incidence of recovery and patterns of antibiotic susceptibility of pathogenic bacteria vary between intensive care units (ICUs) in a single teaching hospital. METHODS: Culture and susceptibility results were collected prospectively for a 3-month period (April through June 1999) in each of the surgical, trauma, and medical ICUs. The number of unique isolates and susceptibility patterns were determined. Susceptibility of isolates among ICUs was compared with chi2. RESULTS: Statistically significant differences between ICUs in susceptibility to various antibiotics were found for Staphylococcus aureus, Enterococcus sp, Acinetobacter sp, Enterobacter sp, Klebsiella sp, and Pseudomonas sp. Notably, vancomycin resistant Enterococcus was not seen in the medical ICU, whereas it was seen in both the surgical and trauma ICUs. Klebsiella spp resistant to ceftazidime were seen only in the trauma ICU. The aminoglycosides and quinolones had attenuated activity against Pseudomonas sp in the surgical ICU, whereas they remained highly effective in the trauma ICU. Cefazolin had no activity against the Enterobacter sp in either of the surgical ICUs, but was highly effective in the medical ICU. CONCLUSION: Although the microbiologic results of this study should not be extrapolated to other institutions, the principle is of value. There is variability between ICUs in a single large teaching hospital in susceptibility of bacterial pathogens to various antibiotics. This may have implications in the design of empiric antibiotic strategies and the planning of the hospital formulary. Hospital wide or composite ICU antibiograms are inadequate for planning empiric therapy in the ICU. PMID- 11038081 TI - Systemic inflammatory response syndrome score at admission independently predicts mortality and length of stay in trauma patients. AB - BACKGROUND: Recent studies have documented that the systemic inflammatory response syndrome (SIRS) score is a useful predictor of outcome in critical surgical illness. The duration and severity of SIRS are associated with posttrauma multiple organ dysfunction and mortality. We sought to determine whether the severity of SIRS at admission is an accurate predictor of mortality and length of stay (LOS) in trauma patients. METHODS: Prospective data of 4,887 trauma admissions to a Level I trauma center over a 18-month period (January 1997 to July 1998) were analyzed. Patients were stratified by age and Injury Severity Score (ISS), and a SIRS severity score (1 to 4) was calculated at admission (1 point for each component present: fever or hypothermia, tachypnea, tachycardia, and leukocytosis). The SIRS score was evaluated as an independent predictor of mortality and LOS by chi2 and multivariate logistic regression. RESULTS: Trauma patients (n = 4,887, 83% blunt injuries, 72% male) had the following characteristics: 73.1% were age 18 to 45 years, 17.5% were age 46 to 65 years, and 9.4% were age > or =66 years; 77.7% had ISS less than 15, 18.8% had ISS 16 to 29, and 3.5% had ISS greater than 29. Analysis of variance adjusting for age and ISS determined that SIRS score of 2 was a significant predictor of LOS. Furthermore, the relative risk of death increased significantly with SIRS score of 2 when age and ISS were held constant. CONCLUSION: Logistic regression analysis confirmed that a SIRS score of 2 was a significant independent predictor of increased mortality and LOS in trauma patients. These data suggest that admission SIRS scoring in trauma patients is a simple tool that may be used as a predictor of outcome and resource utilization. PMID- 11038082 TI - Relationship of cerebral perfusion pressure and survival in pediatric brain injured patients. AB - BACKGROUND: Adult brain injury studies recommend maintaining cerebral perfusion pressure (CPP) above 70 mm Hg. We evaluated CPP and outcome in brain-injured children. METHODS: We retrospectively reviewed the hospital courses of children at two Level I trauma centers who required insertion of intracranial pressure (ICP) monitors for management of traumatic brain injury. ICP, CPP, and mean arterial pressure were evaluated hourly, and means were calculated for the first 48 hours after injury. RESULTS: Of 188 brain-injured children, 118 had ICP monitors placed within 24 hours of injury. They suffered severe brain injury, with average admitting Glasgow Coma Scale scores of 6 +/- 3. Overall mortality rate was 28%. No patient with mean CPP less than 40 mm Hg survived. Among patients with mean CPP in deciles of 40 to 49, 50 to 59, 60 to 69, or 70 mm Hg, no significant difference in Glasgow Outcome Scale distribution existed. CONCLUSION: Low mean CPP was lethal. In children with survivable brain injury (mean CPP > 40 mm Hg), CPP did not stratify patients for risk of adverse outcome. PMID- 11038083 TI - Stapled versus hand sewn anastomoses in patients with small bowel injury: a changing perspective. AB - INTRODUCTION: Recent studies indicate that trauma patients with hollow viscus injuries requiring anastomosis who are managed with stapling have a higher rate of complications than do those in whom a hand-sewn anastomosis is used. We undertook this study to determine whether this finding applied to patients with small bowel trauma at our institution. METHODS: Records of patients with small bowel injuries were retrospectively reviewed. Demographics, severity of injury, injury management, and outcome data were collected. RESULTS: Patients who had their small bowel injuries managed by hand-sewn repair versus resection and stapled anastomosis demonstrated a nonsignificant decrease in overall complication rate (35% vs. 44%) and rate of intra-abdominal complication (10% vs. 18%). Yet the rate of intra-abdominal abscess formation was significantly lower with hand-sewn repair than with resection and stapled anastomosis (4% vs. 13%). However, when hand-sewn primary repairs were excluded from the analysis and injuries that required resection and either stapled or hand-sewn anastomosis were compared, there was a similar overall complication rate (41% vs. 41%) and rate of intra-abdominal complications (17% vs. 21%). CONCLUSION: The rate of intra abdominal complications did not differ significantly between patients requiring small bowel resection and reanastomosis managed by either a stapled or hand-sewn technique. In our experience, surgical stapling devices appear to be safe for use in repairing traumatic small bowel injury. PMID- 11038084 TI - Enteral nutritional support and wound excision and closure do not prevent postburn hypermetabolism as measured by continuous metabolic monitoring. AB - BACKGROUND: Estimation of nutritional needs in burn patients is difficult. In 24 severely burned patients, we measured CO2 production and O2 consumption continuously during their period of mechanical ventilation. METHODS: Patients with extensive burns were placed on a continuous metabolic monitor (CMM) (Puritan Bennett Co., Kingwood, TX), and metabolic expenditure was recorded each 24 hours. High protein enteral feedings were started within several hours of admission, and administration rates were adjusted to meet daily caloric demands as determined by the CMM. Full-thickness wounds were excised as early as patient condition permitted, and wounds were closed with autograft, allograft, or TransCyte (Advanced Tissue Sciences Inc., La Jolla, CA). Daily 24-hour caloric needs as measured by CMM were compared with baseline caloric needs predicted by the Harris Benedict equation and also compared with actual daily caloric intake. Patients were removed from study when they were off continuous mechanical ventilation. RESULTS: A total of 24 patients were studied, with a mean age of 46 years and a 44% total body burn size (partial- and full-thickness). All full-thickness burns were completely excised by a mean of 6.5 days postburn. Mean daily energy expenditures remained elevated through the duration of the study period (42 days), with a mean elevation of 184.9% of baseline as predicted by Harris Benedict equation. Patients received enteral feedings, which met 99.4% of actual caloric needs as predicted by CMM during the study period. CONCLUSION: Continuous metabolic monitoring demonstrates that early wound excision and wound closure, coupled with aggressive enteral nutritional support with high protein formulas, do not prevent the marked hypermetabolism that accompanies thermal injury. PMID- 11038085 TI - Influence of glucose kinetics on plasma lactate concentration and energy expenditure in severely burned patients. AB - BACKGROUND: In critically ill patients, elevation in the plasma lactate concentration has traditionally been interpreted as indicating a deficiency in oxygen availability and is often an impetus to increase oxygen delivery clinically. However, another possible basis for increased lactate concentrations may be simply a mass effect from increased pyruvate availability (i.e., accelerated glycolysis). METHODS: In six hypermetabolic burned patients, the rates of glucose production and oxidation were quantified using a tracer infusion of 6,6 d2 glucose combined with indirect calorimetry. Measurements were obtained after a 9-hour fast and after a 3-hour infusion of unlabeled glucose at 30 micromol/kg/min. No patient was overtly septic, hypoxic, or hypovolemic. RESULTS: The infusion of glucose significantly increased the arterial glucose concentration and rate of glucose oxidation, with a corresponding increase in the arterial plasma concentration of lactate and pyruvate. Resting energy expenditure and oxygen consumption were not affected by the infusion of glucose. CONCLUSIONS: These findings show that elevations in plasma lactate in severely injured patients may, in part, be related to increases in glucose flux and not entirely a reflection of any deficit in oxygen availability. Such findings highlight a potential pitfall for interpreting plasma lactate concentrations as an index of tissue oxygen availability in hypermetabolic patients. PMID- 11038086 TI - Accuracy of administrative data in trauma: splenic injuries as an example. AB - BACKGROUND: Accurate data are needed to evaluate clinical outcomes, therapeutic modalities, and quality of care in trauma. Administrative data, usually used for billing, have been used to evaluate performance and assess therapy in other medical specialties. This study was performed to determine whether administrative databases are accurate in the recording of information about trauma patients with splenic injuries. METHODS: Patients who had blunt splenic injuries were identified using a state trauma registry. The medical records of those patients were reviewed. The data collected by chart review were compared with data in the statewide administrative database of patients who had splenic injuries at the same four Level I and II trauma centers in the same 5-year period. Age, sex, admission date, and hospital were matched to assure comparison of the identical cohort. chi2 analysis was used to compare dichotomous data and Student's t test continuous data. RESULTS: The administrative database identified 641 and the trauma registry identified 529 patients with a diagnosis of splenic injury. A total of 401 patients were found in both databases. Of these, 120 (22.7%) patients were not recorded in the administrative database. Injury Severity Score was underreported by the administrative database (25.74 +/- 14.7 vs. 19.52 +/- 11, p < 0.0001). The administrative database underreported orthopedic, chest, and head injuries (317 vs. 215, 325 vs. 228, and 234 vs. 155, respectively; all p < 0.0001). Use of abdominal computed tomographic scan and diagnostic peritoneal lavage were also underreported (260 vs. 56 and 104 vs.17, both p < 0.0001). The number of operations on the spleen and number of orthopedic procedures were underreported (259 vs. 225, p < 0.014 and 147 vs. 94, p < 0.0001). Complications were markedly underreported by the administrative database (200 vs. 47, p < 0.0001) CONCLUSION: This study shows that administrative data lack accuracy in the recording of associated injuries, injury severity, diagnostics, procedures, and outcomes data in patients with splenic injuries. Whether these data should be used to evaluate treatment modalities or quality of care in trauma is questionable. PMID- 11038087 TI - Helical computed tomographic scan in the evaluation of mediastinal gunshot wounds. AB - BACKGROUND: The standard evaluation of mediastinal gunshot wounds usually requires angiography and either esophagoscopy or esophagography. In the present study, we have evaluated the role of helical computed tomographic (CT) scanning in reducing the need for angiographic and esophageal studies. METHODS: This was a prospective study of patients with mediastinal gunshot wounds who were hemodynamically stable and would otherwise require angiography and esophageal evaluation. All patients underwent CT scan of the chest with intravenous contrast to delineate the missile trajectory. If the missile tract was in close proximity to the aorta, great vessels, or esophagus, then traditional evaluation with angiographic or esophageal evaluation was pursued. RESULTS: A total of 24 patients met the inclusion criteria and underwent CT scan evaluation of their mediastinal gunshot wounds. One patient was taken for sternotomy to remove a missile embedded in the myocardium solely on the basis of the result of the CT scan. Because of proximity of the bullet tract, 12 patients required additional evaluation with eight angiograms and nine esophageal studies. One of these patients had a positive angiogram (bullet resting against the ascending aorta) and underwent sternotomy for missile removal; all other studies were negative. The remaining 11 patients were found to have well-defined missile tracts that approached neither the aorta nor the esophagus, and no additional evaluation was pursued. There were no missed mediastinal injuries in this group. Overall, 12 of 24 patients (50%) had a change in management (either received an operation or avoided additional radiographic or endoscopic evaluation) on the basis of the CT scan. CONCLUSION: The helical CT scan provides a rapid, readily available, noninvasive means to evaluate missile trajectories. This permits accurate assessment of potential mediastinal injury and reduces the need for routine angiographic and esophageal studies. PMID- 11038088 TI - Blunt and penetrating trauma of the thoracic aorta and aortic arch branches: an autopsy study. AB - BACKGROUND: Due to the highly lethal nature of trauma of the thoracic aorta and aortic arch branches (TA-AAB), autopsy studies are essential for the investigation of its epidemiologic characteristics. METHODS: The reports of 11,446 consecutive medicolegal autopsies were reviewed. Among 1,980 injury related fatalities, 251 victims (12.7%) with 302 TA-AAB injuries were found. Several trauma variables were recorded and their relations were examined. RESULTS: Blunt TA-AAB injuries were recorded in 86.4% of the victims. They were located mainly at the aortic isthmus and distal descending thoracic aorta and were accompanied to a great extent by extrathoracic trauma. The vast majority of penetrating lacerations were located at the ascending aorta, arch, and arch branches and were mostly associated with other lethal intrathoracic injuries. All penetrating trauma victims died before reaching the hospital, whereas 5.5% of the blunt trauma victims were admitted to the hospital alive. CONCLUSION: Major differences between blunt and penetrating TA-AAB injuries were revealed, regarding their location, patterns of concomitant injuries, and victims' survival time. Patients injured in motor vehicle crashes, as opposed to various other causes of trauma, were found to have the best chances of reaching the hospital alive. PMID- 11038089 TI - Assessment of gunshot bullet injuries with the use of magnetic resonance imaging. AB - BACKGROUND: Magnetic resonance imaging (MRI) is rarely used for preoperative assessment of shotgun injuries because of concerns of displacing the possibly ferromagnetic foreign body within the surrounding tissue. METHODS: A total of 56 different projectiles underwent MRI testing for ferromagnetism and imaging quality in vitro and in pig carcasses with a commercially available 1.5-MRI scan. Image quality was compared with that of computed tomographic scans. RESULTS: Projectiles with ferromagnetic properties can be distinguished easily from nonferromagnetic ones by pretesting the motion of an identical projectile within the MRI coil. When ferromagnetic projectiles were excluded, MRI yielded the more precise images compared with other imaging techniques. Projectile localization and associated soft tissue injuries were visualized without artifacts in all cases. CONCLUSIONS: When ferromagnetic foreign bodies are excluded by pretesting their properties within the MRI with a comparative projectile, MRI portends an excellent imaging procedure for assessing the extent of injury and planning the removal by surgery. PMID- 11038090 TI - Glucocorticoid and Fas ligand induced mucosal lymphocyte apoptosis after burn injury. AB - BACKGROUND: The purpose of this study was to examine the effects of a steam burn injury on apoptosis in gut-associated lymphoid tissue and to determine whether endogenous glucocorticoid and Fas ligand signaling were involved in this process. METHODS: Histologic analysis, in situ deoxynucleotidyl transferase dUTP nick-end labeling staining and annexin V and 7-amino-actinomycin-D flow cytometry of lymphocyte populations were evaluated in intraepithelial lymphocytes and Peyer's patch. Additional mice were pretreated with a glucocorticoid receptor antagonist (mifepristone) before the steam burn. Similarly, C3H/HeJ-FasL(gld) mice lacking functional Fas ligand were also studied. RESULTS: Apoptosis was significantly increased in intraepithelial lymphocytes and Peyer's patch after the burn injury. Mifepristone pretreatment significantly reduced apoptosis in both T- and B-cell populations in intraepithelial lymphocytes after the burn injury. In contrast, the increased apoptosis seen in B-cells from Peyer's patch was not seen in C3H/HeJFasL(gld) mice, whereas the increased apoptosis in CD8+ T-cells was unaffected. CONCLUSION: Both corticosteroids and FasL contribute to the apoptosis in gut-associated lymphoid tissues early after burn injury. PMID- 11038091 TI - Correction of cubitus varus by French or dome osteotomy: a comparative study. AB - BACKGROUND: The purpose of this study was to compare the outcomes of two different techniques of supracondylar osteotomy, French osteotomy and dome osteotomy, for the correction of posttraumatic cubitus varus. METHODS: A comparative randomized study was undertaken of 25 patients (average age 10 years) with an established posttraumatic cubitus varus deformity (mean time from injury to corrective osteotomy, 1.7 years). Patients were followed-up with for 1 year, and carrying angle, Baumann's angle, and internal rotation deformity were measured. Postoperative complications were assessed. RESULTS: A significant (p < 0.01) correction of carrying angle and Baumann's angle was achieved with both techniques, with no statistically significant differences between them. Although internal rotation deformity was corrected by both techniques, the correction was significantly greater with the dome osteotomy (p < 0.01). However, the persistence of internal rotation did not seem to affect the final outcome. There was a higher incidence of postoperative complications in the dome osteotomy group, including infection (1), inadequate correction (1), nerve palsy (1), loss of motion (5), and circulatory compromise (1). CONCLUSION: The dome osteotomy is technically more difficult than the French osteotomy and has a higher incidence of complications. We suggest that the French osteotomy be used for the correction of cubitus varus after supracondylar fractures of the elbow in children. PMID- 11038092 TI - Blunt hepatic injury: minimal intervention is the policy of treatment. AB - BACKGROUND: Many publications recommend nonoperative treatment for stable blunt hepatic injury patients. Unstable hemodynamic status is the only indication for surgery. When operation is indicated, controversies exist regarding which operative procedure will be more beneficial to the patients. The purposes of this study are to compare the results of operative and nonoperative management of patients with blunt hepatic injuries and to identify the optimal surgical approach when surgery is indicated. METHODS: Different prospective protocols of treating adult blunt hepatic injuries were conducted. From 1992 to 1993 (group I), urgent surgery would be performed in the presence of hemoperitoneum. The policy shifted to aggressive nonoperative approach between 1996 and 1997 (group II). The patients from each period were divided into three subgroups. Group A included the patients who received nonoperative treatment in either period. Group B consisted of the patients who received surgery in the first period and nonoperative management in the second period. Group C included the patients who were operated on in either group. Comparisons were made between matched groups. RESULTS: Groups IA and IIA patients had minor injuries and could be successfully treated nonoperatively. The results of groups IB and IIB were similar concerning hospital stay, morbidity, and mortality. Transfusion requirements of group IIB patients were significantly higher (2.2 vs. 1.1 units,p = 0.01) than those of group IB. However, 25 (58%) celiotomies of group IB patients were nontherapeutic. When surgery was indicated, group IC patients had significantly higher liver related mortality (14 of 49 vs. 3 of 55, p = 0.002). Anatomic resection was performed more frequently in that period. CONCLUSION: Nonoperative treatment significantly decreased the rate of nontherapeutic laparotomy but carried the risks of higher transfusion requirements and delaying operation. When surgery was indicated, the policy of minimal intervention positively affected the patients' outcomes. The goal of surgery should be hemorrhage control rather than resection of the injured liver tissues. PMID- 11038093 TI - Prognostic value of trauma scores in pediatric patients with multiple injuries. AB - BACKGROUND: For the quantification of multiple injuries in children, a range of different trauma scores are available, the actual prognostic value of which has, however, not so far been investigated and compared in a group of patients. METHODS: In 261 polytraumatized children and adolescents, 11 trauma scores (Abbreviated Injury Scale [AIS], Injury Severity Score [ISS], Glasgow Coma Scale [GCS], Acute Trauma Index [ATI], Shock Index [SI], Trauma Score [TS], Revised Trauma Score [RTS], Modified Injury Severity Score [MISS], Trauma and Injury Severity Score [TRISS]-Scan, Hannover Polytrauma Score [HPTS], and Pediatric Trauma Score [PTS]) were calculated, and their prognostic relevance in terms of survival, duration of intensive care treatment, hospital stay, and long-term outcome analyzed. RESULTS: With a specificity of 80%, physiologic scores (TS, RTS, GCS, ATI) showed a greater accuracy (79-86% vs. 73-79%) with regard to survival prediction than did the anatomic scores (AIS, HPTS, ISS, PTS); combined forms of these two types of score (TRISS-Scan, MISS) did not provide any additional information (76-80%). Overall, the TRISS-Scan was the score that showed the highest correlation with duration of treatment and long-term outcome. Trauma scores specially conceived for use with children (PTS, MISS) failed to show any superiority vis-a-vis trauma scores in general. CONCLUSION: With regard to prognostic quality and ease of use in the practical setting, TS and the TRISS Scan are recommended for polytrauma in children and adolescents. Special pediatric scores are not necessary. PMID- 11038095 TI - Abdominal compartment syndrome in patients with isolated extraperitoneal injuries. PMID- 11038094 TI - Sickness Impact Profile Score versus a Modified Short-Form survey for functional outcome assessment: acceptability, reliability, and validity in critically ill patients with prolonged intensive care unit stays. AB - BACKGROUND: Quality of life after surgical critical illness is an important measure of outcome. The Sickness Impact Profile Score (SIP) has been validated in critically ill patients, but the Modified Short-Form (MSF) has not been directly compared with it. METHODS: The SIP and MSF-36 were coadministered to 127 patients (surrogates) with a prolonged surgical critical illness at baseline at 1, 3, 6, and 12 months. Reliability, validity, and acceptability were determined for overall and subscores at each time point. RESULTS: The overall SIP and eight subscores, including physical health and psychosocial health, were all significantly improved at 1 year compared with baseline (p < 0.05). However, the MSF-36 was improved only in health perception (p < 0.05), but pain scores were higher (p < 0.05) than at baseline. Internal consistency of the MSF-36 was poor at 1 and 3 months. Correlation between the tools was excellent at baseline and 1 year but variable in overall and subscores at other time points. CONCLUSION: The SIP is more comprehensive, reliable, and acceptable in determining specific quality-of-life abnormalities, but the MSF-36 is easier to administer and correlates well at baseline and 1 year in patients with a prolonged critical illness. PMID- 11038096 TI - Avulsion of the pubic branch of the inferior epigastric artery: a cause of hemodynamic instability in minimally displaced fractures of the pubic rami. PMID- 11038097 TI - Transverse sacral fracture with intrapelvic intrusion of the lumbosacral spine: case report and review of the literature. PMID- 11038098 TI - Bilateral below-knee amputation surgery at the scene: case report. PMID- 11038099 TI - Neutropenic enterocolitis in a trauma patient during antibiotic therapy for osteomyelitis. PMID- 11038100 TI - Traumatic dislocation of the mandibular condyle into the temporal fossa in a child. PMID- 11038101 TI - Quadriplegia after upper extremity trauma: case report. PMID- 11038102 TI - Lacerations of the left ventricle from rib fractures after blunt trauma. PMID- 11038103 TI - Air pellet embolization after penetrating cardiac injury. PMID- 11038104 TI - Blunt traumatic azygous vein injury diagnosed by computed tomography: case report and review of the literature. PMID- 11038105 TI - Combined penetrating injury of the innominate artery, left common carotid artery, trachea, and esophagus. PMID- 11038106 TI - Treatment of bilateral blunt carotid injury using percutaneous angioplasty and stenting: case report and review of the literature. PMID- 11038107 TI - Successful recovery after an orbitocranial injury. PMID- 11038108 TI - Taking care of the "good guys": a trauma center-based model of medical support for tactical law enforcement. PMID- 11038109 TI - Taking care of the "good guys": a trauma center-based model of medical support for tactical law enforcement. PMID- 11038110 TI - Evaluate the complications of prone positioning in patients with severe ARDS after trauma. PMID- 11038111 TI - Posterolateral dislocation of the elbow with concomitant fracture of the lateral humeral condyle. PMID- 11038112 TI - Extraordinary case of shoulder trauma caused by a foreign bone. PMID- 11038113 TI - Clear and insightful review of hepatic venous injuries. PMID- 11038114 TI - Modelling of the solvent evaporation method for the preparation of controlled release acrylic microspheres using neural networks. AB - The purpose of the present study was to model the solvent evaporation procedure for the preparation of acrylic microspheres by using artificial neural networks (ANNs) to obtain an understanding of the selected preparative variables. Three preparative variables, the concentration of the dispersing agent (sucrose stearate), the stirring rate of emulsion system, and the ratio of polymers (Eudragit RS-L) were studied, each at different levels, as input variables. The response (output) variables examined to characterize microspheres and drug release were the size of the microspheres and T63.2%, the time at which 63.2% of drug is released. The results were also analysed by the multiple linear regression (MLR) to provide a comparison with the ANN methodology. Although both ANN and MLR methods were found to be similar in characterizing the process studied, the results showed that an ANN method gave a better prediction than the MLR method. For the size values of the microspheres, the predictability of the ANN model was quite high (R2 = 0.9602) based on the input variables. A relationship between these variables and size values of microspheres was also obtained by the MLR model (R2 = 0.9050). The performances of both models for the release data from microspheres based on the same input variables were at the level of 53%. According to the results, the ANN methodology can provide an alternative to the traditional regression methods, as a flexible and accurate method to study process and formulation factors. PMID- 11038115 TI - Effect of tabletting compaction pressure on alginate microspheres. AB - Alginate and alginate-hydroxypropylmethylcellulose (HPMC) microspheres were prepared by the emulsification method. The compaction of microspheres for producing tablet dosage forms raises concerns about possible damage to microsphere walls with subsequent unpredictable dissolution rates. The effect of different compaction pressures on the integrity of the microspheres was investigated. The addition of a diluent, microcrystalline cellulose (MCC), was required to make compacts containing alginate and alginate-HPMC microspheres. Compacts containing alginate-HPMC (7:3) microspheres had the highest crushing strength followed by compacts containing alginate-HPMC (9:1) microspheres and alginate microspheres. However, compact crushing strength did not vary significantly with increased compaction pressures over the range of compaction pressures investigated. Differences in the drug release profiles of the original non-compacted and compacted alginate and alginate-HPMC microspheres were slight and not marked. Although dentation and distortion of the microspheres were observed with increasing compaction pressures, the microspheres generally remained intact, with minimal rupture/fracture. PMID- 11038116 TI - Sustained release properties of alginate microspheres and tabletted microspheres of diclofenac sodium. AB - This study focused on the properties of diclofenac sodium (DNa) alginate (alg) microspheres and tabletted DNa alg microspheres using different polymers as additives. DNa alginate microspheres were prepared by the emulsification method and different polymers such as Eudragit (Eud) NE 30 D, Eudragit (Eud) RS 30 D and Aquacoat, which were incorporated into alg gel to control the release rate of drug. The release properties of DNa alg microspheres (1:1) were affected by the size, drug load of microspheres and also by the incorporated polymers, pH and ionic strength of dissolution medium. Tabletting of alg microspheres using carrageenan (carr), alg, pectin, NaCMC, tragacanth (trgh) and HPMC as additives in a (50:50) ratio produced tablets with good physical properties and also better controlled release of DNa. Dissolution studies were carried out in pH 7.2 phosphate buffer and phosphate buffers whose pH values were gradually changed from pH 3 to 7.4. The rank order of DNa release from tablets was carr < alg < pectin < NaCMC < trgh < HPMC which relates to the viscosity and swelling properties of polymers. The drug release was very slow from trgh and HPMC based tablets, but addition of carr or alg in different ratios could adjust the release rate of drug. PMID- 11038117 TI - In vitro degradation and dissolution behaviours of microspheres prepared by three low molecular weight polyesters. AB - Three low-molecular weight polyesters, poly(L-lactic acid) (PLA), copoly(lactic acid/glycolic acid) (PLGA) and poly(delta-valerolactone) (PV), were used to prepare water-soluble sodium diclofenac-loaded microspheres by using the oil-in oil (o/o) emulsification-solvent evaporation method. Their micromeritic and physicochemical properties, and degradation and dissolution behaviours were determined in vitro. The results indicate that high encapsulation efficiency and better monodispersity might be achieved by the o/o emulsification-solvent evaporation method, depending on the amount of drug loading used. The slower evaporation of organic solvent from the system during microencapsulation seemed to modify the crystallinity of drug and polyester in the microspheres, determined by powder x-ray diffractometry and differential scanning calorimetry. The in vitro degradation rate of all the microspheres in pH7.4 phosphate buffer solution showed first-order kinetics and ranked in the order of PLGA > PLA > PV microspheres. Furthermore, the first-order release rate was also found in all the microspheres after an initial drug burst and ranked in the order of PLGA> PLA > PV microspheres, too. The relationship between degradation and dissolution behaviours of these microspheres is discussed. PMID- 11038118 TI - Branched oligoester microspheres fabricated by a rapid emulsion solvent extraction method. AB - Methyl formate was used as the solvent of biodegradable oligoesters for the fabrication of microspheres with encapsulated bovine serum albumin (BSA). The procedure of dispersion of the double emulsion of the w/o/w type and its dilution and solvent extraction is very rapid, taking only several minutes. A higher yield and better encapsulation efficiency were obtained with copolymers of DL-lactic acid with mannitol than with pure linear poly DL-lactic acid. The procedure was accelerated, and yields and encapsulation efficacy were enhanced by the addition of 5% methyl formate to the external water phase. The microspheres were smaller than 100 microm. No benefits were obtained from the addition of wetting agents or other additives to the intermediate (oligoesteric) phase. Further development should concentrate particularly on hydrodynamic conditions and optimization of the composition of the external phase. PMID- 11038119 TI - Biodegradable nanoparticles as a delivery system for cyclosporine: preparation and characterization. AB - Cyclosporine (CyA) was incorporated into polycaprolactone nanoparticles (PCL-NP) in order to increase its oral bioavailability and to control drug distribution, thereby potentially reducing its toxicity. Prior to in vivo studies, the carrier was optimized and characterized by using different techniques. Light scattering (LS) and transmission and scanning electron microscopy (TEM and SEM) indicated the NP were spherical in shape with a mean size of approximately 100 nm. The influence of the solvent evaporation conditions and the polymer and drug amounts on CyA incorporation was established in order to optimize drug loading. When acetone and excess water were removed at constant temperature, no aggregation phenomena were observed. A value of 180 mg PCL was the minimum polymer amount necessary to encapsulate 95% of the drug initially added to the preparation. Under these conditions, HPLC analysis revealed that approximately 130 microg CyA per mg PCL were incorporated for a total CyA concentration of 2.5 mg/ml, being part of the drug adsorbed onto the particle surface. No structural changes or instability of the components during NP preparation were detected by gel permeation chromatography (GPC) and differential scanning calorimetry (DSC). However, GPC studies showed a competition between poloxamer and CyA for adsorption onto the carrier. In addition, DSC results suggested that at least part of the drug associated to NP remained in its crystal form. Therefore, CyA loaded NP were easily manufactured and characterized and allow for the administration of therapeutic drug doses to experimental animals. PMID- 11038120 TI - Labile conjugation of a hydrophilic drug to PLA oligomers to modify a drug delivery system: cephradin in a PLAGA matrix. AB - The physical entrapment of a hydrophilic drug within degradable microspheres is generally difficult because of poor entrapment yield and/or fast release, depending on the microsphere fabrication method. In order to counter the effects of drug hydrophilicity, it is proposed to covalently attach the drug to lactic acid oligomers, with the aim of achieving temporary hydrophobization and slower release controlled by the separation of the drug from the degradable link within the polymer matrix. This strategy was tested on microspheres of the antibiotic cephradin. As the prodrug form, the entrapment of the drug was almost quantitative. The prodrug did degrade in an aqueous medium, modelling body fluids, but cleavage did not occur at the drug-oligomer junction and drug molecules bearing two lactyl residual units were released. When the prodrug is entrapped within a PLAGA matrix, no release was observed within the experimental time period. However, data suggest that conjugation via a bond more sensitive to hydrolysis than the main chain PLA ester bonds should make the system work as desired. PMID- 11038121 TI - 5-Fluorouracil-loaded chitosan coated polylactic acid microspheres as biodegradable drug carriers for cerebral tumours. AB - The development of injectable microspheres for anticancer drug delivery into the brain is a major challenge. The possibility of entrapping 5-fluorouracil (5-FU) in chitosan coated monodisperse biodegradable microspheres with a mean diameter of 10-25 um was demonstrated. An emulsion of 5-FU (in water) and polylactic acid (PLA) dissolved in acetone-dichloromethane mixture was poured into an aqueous solution of chitosan (or poly-vinyl alcohol) with stirring using a high-speed homogenizer, for the formation of microspheres. 5-FU recovery in microspheres ranged from 44-66% depending on the polymer and emulsification systems used for the preparation. Scanning electron microscopy revealed that the chitosan coated microspheres had less surface micropores compared to PVA based preparations. The drug release behaviour from microspheres suspended in phosphate buffered saline exhibited a biphasic pattern. The amount of drug release was much higher initially (approximately 25%), followed by a constant slow release profile for a 30 days period of study. This chitosan coated PLA/PLGA microsphere formulation may have potential for the targeted delivery of 5-FU to treat cerebral tumours. PMID- 11038122 TI - Effect of solid lipid nanoparticles (SLN) on cytokine production and the viability of murine peritoneal macrophages. AB - The purpose of this study was to investigate possible immunomodulatory and cytotoxic effects of solid lipid nanoparticles (SLN) on murine peritoneal macrophages. Immunomodulatory effects of SLN composed of either a lipid- (glycerol-behenate) or a wax (cetylpalmitate) matrix stabilized by the surfactant Poloxamer 188 were analysed by detection of proinflammatory and down-regulatory cytokines in supernatants of thioglycollate-elicited peritoneal macrophages using enzyme-linked immunosorbent assay (ELISA). Cytotoxicity of SLN was assessed using the ITT test. Incubation of macrophages with either SLN at low concentrations did not increase production of interleukin (IL)-6, IL-12, and tumour necrosis factor (TNF)-alpha. At higher SLN concentrations, a concentration-dependent decrease in IL-6 secretion was observed compared to background production of IL-6 by untreated macrophages. IL-12 and TNF-alpha production was neither detected in supernatants of macrophages treated with SLN at any concentration nor in those of untreated cells. The decrease in IL-6 secretion was paralleled by concentration dependent cytotoxicity of SLN on these cells. In contrast, incubation with polystyrene reference particles neither resulted in decreased IL-6 production nor in a loss of viability. SLN-treated macrophages were found to up-regulate their cytokine production following stimulation with Pansorbin, despite the concentration-dependent cytotoxicity induced by SLN. Down-regulatory effects on SLN-treated macrophages by IL-10 were not observed. In conclusion, incubation of SLN with murine peritoneal macrophages did not induce the production of proinflammatory and down-regulatory cytokines. At high concentrations of SLN, cytotoxic effects on these cells were observed. Cytotoxicity appears to be the main cause of decreased cytokine production by these cells. PMID- 11038123 TI - Plastic behaviour of polyelectrolyte microcapsules derived from colloid templates. AB - The deformability and osmotic properties of hollow microcapsules were studied by means of the micropipette video microscopic technique. The microcapsules were prepared by consecutive multiple adsorption of the polyanion, poly(styrene sulphonate), and the polycation, poly(allylamine hydrochloride), onto melamine formaldehyde resin latex of 5 microm diameter, which was decomposed after completing the coating by transferring to hydrochloric acid of pH 1.1. The polyelectrolyte microcapsules reacted to micropipette suction with plastic deformation. If lipids are added to the polyelectrolyte layers, the capsules cannot be visibly deformed by micropipette suction up to 10(4) N/m2. However, plastic shrinking was observed if the stress was generated by the osmotic pressure of a sucrose solution of 10(6) N/m2. PMID- 11038124 TI - Literature alerts. PMID- 11038125 TI - Experimental evidence of host specificity of Bartonella infection in rodents. AB - A large number of Bartonella species and genetic variants were compared for their ability to cause bacteremia in different rodent species: the cotton rat (Sigmodon hispidus), white-footed mouse (Peromyscus leucopus), BALB/c mouse and Wistar rat. Experimental data supported field observations that host specificity can occur among certain Bartonella species and rodent species. Bacteremia could only be readily produced in cotton rats or white-footed mice if the strains used for inoculation were originally obtained from the same species or from a phylogenetically close species. A few Bartonella colonies could be observed in the blood of some BALB/c mice by 7 days after inoculation, but no evidence of the persistence of the infection was found. Host specificity suggests the possibility of a long co-speciation of Bartonella species with their rodent hosts. Host parasite relationships measured by the duration and level of bacteremia and the minimal infectious dose may serve as additional criteria for classification of Bartonella isolates obtained from natural environments. PMID- 11038126 TI - Anaplasma marginale inactivated vaccine: dose titration against a homologous challenge. AB - The present study was performed to dose-titrate an Anaplasma marginale experimental immunogen derived from partially purified initial bodies from three geographically different Mexican strains. Three five-bovine groups were inoculated twice on days zero and 21 with A. marginale initial bodies equivalent to 1.5 x 10(10) (group I), 3 x 10(10) (group II) or 6 x 10(10) (group III) infected erythrocytes mixed with STDCM adjuvant. A similar group served as non vaccinated controls. All four groups were challenged with 1 x 10(8) infected erythrocytes from a donor cow with an increasing rickettsemia of strain MEX-15 on day 87 post-vaccination. The prepatent period was very similar for all four groups. All five non-vaccinated controls presented typical acute anaplasmosis syndrome reaching a mean of 30.9% rickettsemia and a loss of 73.4% in the packed cell volume (PCV). Two of five controls died of acute anaplasmosis. Within the vaccinated groups only one animal (group II) suffered acute disease and died. Although all the other vaccinated animals were free of clinical signs, they developed very low rickettsemias (3.2, 3.8 and 4.3%) and PCV losses of 49.9, 47.8, and 49.3% for groups I, II and III. The starting mean weight was very similar for all four groups. All animals lost weight following challenge but losses for groups I and II were lower and significantly different from group IV losses (P < or = 0.1). Although there were no significant differences among vaccinated groups, group III was more severely affected. Taken altogether, these results show a 93.3% protection against both illness and death for all groups; and 100% protection for groups I and III, and 80% for group II. PMID- 11038127 TI - Characterization of Salmonella isolates from beef cattle, broiler chickens and human sources on Prince Edward Island. AB - Non-typhoid Salmonella serovars remain a potential threat to human health, and beef cattle and broiler chickens are possible sources of these organisms on Prince Edward Island (PEI). In this study, the ceca of beef cattle belonging to fasted and non-fasted groups, and broiler chickens were examined for Salmonella at the time of slaughter. The characteristics of the isolates, including antimicrobial resistance patterns and virulence genes, were studied along with the isolates obtained from cases of human salmonellosis on PEI during the study period (1996-97). The prevalence of Salmonella in beef cattle was 4.6% (11/240). The rate was significantly higher in fasted cattle (7.46%), than in non-fasted cattle (0.94%). The prevalence rate in chickens was 32.5% (39/120). In beef cattle, Salmonella typhimurium phage type (PT) or definitive type (DT) 104 which was resistant to ampicillin, chloramphenicol, streptomycin, sulfisoxazole and tetracycline, was the most predominant type (64%). In chickens, S. heidelberg, with resistance to gentamicin, streptomycin and sulfisoxazole, predominated. Of 26 isolates from humans, the most common serovar was S. typhimurium, including a multidrug-resistant strain of DT104. Examination by PCR revealed presence of the virulence gene invA in all serovars, and the spvC gene in all S. typhimurium isolates, of both beef cattle and human origin. Among the other serovars the latter gene was found in 7 human isolates, but in none of the chicken or beef isolates. All but 3 of the spvC-positive isolates possessed a 90 kilobasepair (kbp) plasmid suggesting that the 3 isolates had the spvC gene on their chromosome. These findings were confirmed by plasmid DNA isolation using 3 different protocols and by sequence analysis of the spvC-PCR product. PMID- 11038128 TI - Characterization of shiga toxin-producing Escherichia coli strains isolated from calves in Poland. AB - Fecal samples from 67 3-5-months-old calves with diarrhea were screened for the presence of shiga toxin-producing Escherichia coli (STEC). Several accessory virulence factors genes were also tested. Among 192 E. coli isolates tested, 15 (7.6%) were found to harbour the shiga toxin 1 or 2 (stx1 or stx2) genes. The stx2-carrying samples were further subtyped by PCR for the stx2c, stx2d, and stx2e toxin variants. It was shown that stx2-positive bacteria mainly possessed the stx2c shiga toxin type gene. The enterohemolysin (hlyA) and intimin (eae) genes were found in seven (46.7%) STEC strains whereas the cytotoxic necrotizin factor 1 and 2 or the P fimbrial genes were detected in two isolates only. This study confirmed that calves are a reservoir of STEC strains (with all pathogenicity genes) that may be virulent for humans. PMID- 11038129 TI - Detection and characterization of Clostridium species in soil of Zambia. AB - In the retrospective study of soil-borne diseases of cattle in Zambia, malignant edema and blackquarter were widespread. One hundred and sixty-five cases with malignant edema and 103 cases with blackquarter were reported between 1985 and 1997. It was found that specific soil-conditions associate the emergence of the soil-borne diseases. Soil samples from five areas in Zambia were examined for the presence of genus Clostridium. Direct immunofluorescent assay (IFA) examination showed that C. septicum, C. novyi and C. chauvoei were detected in the soil of specific areas in Zambia, respectively. Causal organisms such as C. perfringens were isolated from the soil samples. The information of area-specific distribution of Clositridium species may give an efficient program in protecting cattle and man. PMID- 11038130 TI - The Irish brawn drain: English League clubs and Irish footballers, 1946-1995. AB - This paper draws on world systems and resource dependency theories to show how the changing recruitment practices of English League clubs have deepened the brawn drain from Irish football, thereby compounding its underdevelopment. An analysis of the origins, method of recruitment and destinations of Irish players (North and South) who appeared in the English League between 1946 and 1995 shows that English clubs imported large numbers of Irish players throughout the second half of the twentieth century. However, it was the inclusion of Irish teenagers within the youth policies of the largest clubs in the period after the 1970s that marked a break from the traditional pattern of buyer-supplier relations. Instead of continuing to purchase players who had established reputations within the Irish leagues, English clubs began to hire the most promising schoolboys before they joined Irish sides. As this practice spread, it eventually eliminated a valuable source of income: the selling of players to English clubs. Despite this development it would, however, be inappropriate to view the relationship between the Irish and English football industries as a simple zero sum game as Irish clubs benefit from employing highly trained young players who return home after failing to establish careers in England. PMID- 11038131 TI - Football fandom and post-national identity in the New Europe. AB - Through European club football, we can begin to detect the outlines of a new Europe of competing cities and regions which are being disembedded from their national contexts into new transnational matrices. Focusing on a specific network of Manchester United fans, broadly located in the city of Manchester, this article examines the development of European consciousness among this group of individuals. This consciousness does not consist of a European supranationalism but rather of a new emphasis on the locale of Manchester and an increasing recognition that Manchester United and the city of Manchester must compete autonomously with other major clubs and cities in Europe. PMID- 11038132 TI - 'Bataille's boys': postmodernity, Fascists and football fans. AB - In his analysis of football hooliganism, Anthony King claims to reveal the historical, conceptual scheme young, male supporters draw upon. This 'masculine vision', he states, is similar to that held by the Freikorps. Both groups are said to adhere to modernist notions of masculinity, sexuality and nationhood, reinforced by rituals which maintain boundaries between these 'proper' males and deviant 'others'. Occasionally, football hooligans breach these boundaries in acts of postmodern transgression. King also claims that fans link sex and violence in their imaginations. In this response I examine King's critique of his fellow theorists; challenge his 'Freikorps-Fans' analogy; demonstrate the problem he has in establishing the sex-violence link and question the relevance of his concept of postmodernity. PMID- 11038133 TI - Electoral democracy, revolutionary politics and political violence: the emergence of Fascism in Italy, 1920-21. AB - This study examines the determination of the Italian Fascists' extra parliamentary, para-military, violent strategy. What were the effects of the socialists' political strategy, relying on electoral democracy, on the creation and strategy of the Fascist Action Squads? A comparison among Italy's 69 provinces, based on quantitative and qualitative historical evidence reveals a distinct pattern in the Fascists' violence. They attacked mainly provinces where the Socialists enjoyed the greatest electoral support. This pattern was a product of two historical processes: (a) the threat of the Socialist party to the landlords' economic and political hegemony, and (b) the landlords' tradition of militant anti-worker organization which culminated in their alliance with the Fascists. The Fascists' struggle for, and takeover of, political power was not an immanent historical necessity. It was first and foremost an anti-socialist reaction. It was shaped both 'from below', by the political power and radicalism of the PSI and the para-military capacity of the Fascist Squads; and 'from above', by the active support the Fascists received from the landlords and the state. Supported by organized landlords and blessed with the authorities' benevolence, the Squads were able to destroy - physically and politically - the legitimately constituted provincial governments of the Socialists. The alliance with the landlords determined the Squads' almost exclusive attacks on Socialist provincial strongholds that constituted the greatest threat to the landlords' interests, while provinces dominated by the ruling Liberal party were excluded from the Squads' path of 'punitive expeditions'. PMID- 11038134 TI - On sources and narratives in historical social science: a realist critique of positivist and postmodernist epistemologies. AB - Critics of the interdisciplinary enterprise of historical sociology commonly contend that the narrational accounts of past social phenomena provided by historians are inadequate to the task of theory-building and testing. In support of this negative assessment, opponents will adduce informational deficiencies in the available data (the standard positivist appraisal of historical evidence), or cite the interpretive anarchy that seemingly prevails at the narrative phase of emplotment (the skeptical, postmodernist contention that historiographic texts 'construct' rather than veridically represent the events they artfully contrive to signify). Both of these lines of criticism are unbalanced, and therefore seriously misleading as regards the epistemic foundations of historical sociological inquiry. The 'social authenticity' and 'informational density' of historical evidence does allow for veridical reconstructions of the past, while the reflexive interpretive protocols of source criticism and the sociology of knowledge can be deployed to provide warrant for discriminating arbitrations between competing theories and narratives. The various epistemological deformations in the study of human affairs that have been encouraged by the old idiographic-nomothetic polarity - chronic ahistoricism within the social sciences, the atheoretical predilections of much conventional historiography - are rectifiable through the consolidation of a fully integrated sociological history, a unified and inclusive historical social science. PMID- 11038136 TI - Spencer is dead, long live Spencer: individualism, holism, and the problem of norms. AB - The debate between the advocates of sociological individualism and those of holism has been pervasive in the development of social theory. This debate is often situated in the false problems of sociology, since it is seen as a particular form of the perennial and irresolvable dilemma between social nominalism and realism, as well as between freedom and determinism. Nevertheless, the debate is far from over within contemporary sociology and other social science, as indicated by the resurgence of individualism in rational action theory and its repudiation by holistic social theories. The aim of this paper is to identify some modern variations on this theme as well as to discern certain common tendencies of two seemingly opposite theoretical perspectives, viz. the convergence upon a normative solution to the problem of social order. This convergence is therefore denoted normative convergence between sociological individualism and holistic sociology. PMID- 11038135 TI - The power and limits of ethnonationalism: Palestinians and Eastern Jews in Israel, 1974-1991. AB - The relative standings of four ethnic groups - Muslim Palestinians, Christian Palestinians, Asian-African Jews, European Jews -were compared, using mobility data from 1974 and 1991. The findings show that despite the lack of government support and the prevalence of inexorable discrimination against Israeli Palestinians, they have narrowed the gap with Asian-African Jews in both education and occupational prestige. This finding demonstrates that ideological and political hegemony is not always effective in improving the socio-economic standing of preferred minorities (Asian-African Jews), and that social and economic structures may counterbalance the anti-Palestinian nationalist ideology. The analysis suggests that residential and educational segregation of Palestinians protects them from direct competition with European Jews, whereas Asian-African Jews have to compete with this dominant group in schools, as well as in the labour market. PMID- 11038137 TI - Autopoiesis and socialization: on Luhmann's reconceptualization of communication and socialization. AB - In 1984, Niklas Luhmann published Soziale Systeme in which he applies the idea of autopoiesis (= self-production) to social systems. Abstracted from its biological connotations, the concept of autopoiesis leads to a sharp distinction between different kinds of autopoietic organization, i.e. between life, consciousness and communication. According to Luhmann, the relationship between social systems and human beings cannot be adequately analysed except by taking into account that they are environments for one another. If this theoretical background is accepted, the concepts and theory of socialization need to be revised. Luhmann takes issues with classical notions such as internalization, inculcation, or 'socialization to the grounds of consensus' (Talcott Parsons). After a historical overview of social systems research and general systems theory, it is indicated how communications trigger further communications and realize the autopoiesis of social systems. In the second part of the article, the distinction between social systems and psychic systems is used to discuss issues crucial to socialization theory. Both a revision of the concept of socialization, and lines for an empirical research programme are proposed in accordance with Luhmann's theory of social systems. PMID- 11038138 TI - Cyclin-dependent kinase and protein kinase C inhibitors: a novel class of antineoplastic agents in clinical development. AB - Malignant cells have acquired adaptations, which give them a growth and survival advantage over normal cells. One effect of many of these adaptations is that many cancerous cells are less likely to undergo programmed cell death (apoptosis) and, moreover, are resistant to chemotherapy-induced apoptosis. Other features of neoplastic cells are the loss of regulated or orderly progression through the cell cycle. In normal and cancerous cells, a balance between proapoptotic and antiapoptotic signals exists. Protein kinase C (PKC) is a cellular serine/threonine kinase with a central role in the mediation of mitogenic signals as well as the regulation of antiapoptotic signals. Inhibition of PKC by a novel group of chemical agents (PKC inhibitors) can induce apoptosis in some malignant cell lines, act as differentiating agents, and enhance the effect of cytotoxic chemotherapy. Other kinase inhibitors are potent inhibitors of kinases involved in the control of cell cycle progression (cyclin-dependent kinases [cdks]). Cdk inhibitors are able to induce cell cycle arrest in neoplastic cells and also act as enhancers of chemotherapy-induced apoptosis. The catalytic domain of different classes of kinases (PKC and cdk) share considerable homology. As a result, many kinase inhibitors that act by blocking the catalytic site are not highly specific and may act as inhibitors of both PKC and cdks. Preclinical studies point to potential applications for some of these PKC/cdk inhibitors, and current clinical trials are exploring the role these agents might have in cancer therapy. In this article, we discuss the rationale for the development of this novel class of agents and highlight those drugs, which have shown promise in clinical testing. PMID- 11038139 TI - Comments on an in vitro model of hormonally refractory prostate cancer. PMID- 11038140 TI - Thymidylate synthase and the cell cycle: what should we believe? PMID- 11038141 TI - Tyrosine kinase inhibitors for suppression of tumorigenic growth of Ras transformed cells: PAK-ing in the signals. PMID- 11038142 TI - Androgen deprivation induces selective outgrowth of aggressive hormone-refractory prostate cancer clones expressing distinct cellular and molecular properties not present in parental androgen-dependent cancer cells. AB - PURPOSE: The mechanism of progression of human prostate cancer (CaP) cells under androgen ablation therapy remains unclear. To study the alternative pathways of CaP cell growth under conditions of androgen deprivation, androgen-independent CaP variants were selected and expanded from an androgen-dependent CaP line via an in vitro androgen deprivation treatment. Cellular and molecular properties of these androgen-independent variants were characterized both in vitro and in vivo and compared with those of their parental androgen-dependent cells. METHODS: Androgen deprivation treatment of an androgen-dependent CaP cell line, LNCaP, was carried out by replacing culture medium with RPMI 1640 medium plus 10% charcoal stripped serum. Cells that survived through the androgen deprivation treatment were harvested and expanded in the androgen-deficient culture medium and were designated CL-1. The CL-1 cells were also recultured in androgen-containing medium and designated CL-2. The growth (cell cycle analysis, 3H-thymidine incorporation assay, growth expansion, and colonization efficiency), expression of CaP-associated markers (semiquantitative reverse transcriptase polymerase chain reaction), interaction with endothelial and bone marrow stromal cells, sensitivity to anticancer agents and radiation (growth inhibition), and tumorigenicity of CL-1 and CL-2 cells were determined and compared with these characteristics in parental LNCaP cells. RESULTS: CL-1 and CL-2 cells are fast growing cells when compared with parental LNCaP cells. They were capable of potentiating the growth of endothelial and bone marrow stromal cells in co culture experiments and acquired significant resistance to radiation and to anticancer cytotoxic agents (Taxol paclitaxel, vinblastine, and etoposide). In contrast to the poorly tumorigenic parental LNCaP cells, CL-1 and CL-2 lines proved highly tumorigenic, exhibiting invasive and metastatic characteristics in intact and castrated mice or in female mice within a short period of 3 to 4 weeks. No growth supplements (e.g., Matrigel) were needed. When transfected with the green fluorescence protein (GFP) gene and transplanted orthotopically in the accessory sex gland, extensive metastatic disease from the primary CL tumor could be identified in bone, lymph nodes, lung, liver, spleen, kidney, and brain. Semiquantitative reverse transcriptase polymerase chain reaction analysis revealed a markedly distinct molecular expression profile in the CL lines: overexpression of basic fibroblast growth factor, interleukin-6, interleukin-8, vascular endothelial growth factor, transforming growth factor-beta, epidermal growth factor receptor, caveolin, and bcl-2 messenger RNAs and marked down regulation of E-cadherin, p-53, and pentaerythritol tetranitrate. CONCLUSIONS: Early administration of hormonal therapy after failure of first-line treatment is associated with a profound clonal selection of aggressive AI variants, such as CL 1 and CL-2 lines. These tumor lines, with their parental counterparts, can serve as valuable tools for studying the cellular and molecular mechanisms of CaP progression and metastasis under hormonal therapy. CL-1 and CL-2 offer a unique and reproducible model for the evaluation of drug sensitivity and for other therapeutic modalities for advanced prostate cancer. PMID- 11038143 TI - Cell cycle regulation of the G0/G1 transition in 5-fluorouracil-sensitive and resistant human colon cancer cell lines. AB - PURPOSE: Resistance to 5-fluorouracil (5-FU) has been associated with thymidylate synthase (TS) gene amplification and increased TS protein levels. Increased TS protein expression has also been found to be a significant independent prognostic factor for disease-free survival and overall survival in patients treated with adjuvant 5-FU-based chemotherapy. In these studies and in our prior preclinical studies, TS has been considered a marker of proliferative capacity. The purpose of the current study was to further evaluate the association between TS levels and cell cycle regulation, by investigating cell cycle kinetics in a 5-FU resistant cell line with constitutive overexpression of TS. The influence of increased TS levels on cell cycle progression may provide insight into methods to overcome 5-FU resistance. MATERIALS: 5-FU-sensitive NCI H630(WT) and 5-FU resistant NCI H630(R1) (with 15- to 20-fold higher TS protein levels) were utilized in this investigation to determine the influence of constitutive overexpression of TS on cell cycle kinetics. RESULTS: There was no apparent influence of increased TS levels on cell cycle distribution during asynchronous growth, and both cell lines reach plateau growth phase in 120 hours, arresting in G0/G1 as determined by flow cytometry. In the H630(WT) cells, this G0/ G1 arrest was associated with a 14- to 17-fold reduction in TS activity and protein levels (using the TS-106 monoclonal antibody), whereas in the H630(R1) cells, only a two to fivefold reduction was noted. Flow cytometry analysis utilizing Ki-67 indicated that there was no evidence of a G0 population in the confluent H630(R1), whereas 26% +/- 7% of confluent H630(WT) cells were Ki-67 negative (G0) and the remainder had low Ki-67 signal intensity. Analysis of pRb phosphorylation and p16 and p21 expression suggested that the arrest point for both cell lines was before the point at which Rb phosphorylation takes place, yet the confluent H630(R1) cells had threefold higher p21 than confluent H630(WT) cells. DISCUSSION: These data suggest that the 5-FU-resistant H630(R1) cell lines arrest at a later point in G0/G1 and have a potentially greater capacity for proliferation. PMID- 11038144 TI - An anti-Ras cancer potential of PP1, an inhibitor specific for Src family kinases: in vitro and in vivo studies. AB - BACKGROUND: We previously found that both PAK, a Rac/CDC42-activated Ser/Thr kinase, and its binding partner PIX are required for malignant transformation caused by oncogenic Ras mutants, such as v-Ha-Ras. Furthermore, oncogenic Ras requires an autocrine pathway to activate PAK. This pathway involves at least two distinct receptor kinases: EGF receptor (ErbB1) and ErbB2. Interestingly, both of these kinases are known to activate Src family kinases that phosphorylate CAT, another binding partner of PIX. PURPOSE: The major aim of this study was to determine whether Src family kinases are required for both Ras-induced PAK activation and malignant transformation. For this purpose, we used PP1, an inhibitor specific for Src family kinases, which does not inhibit either EGF receptor or ErbB2. METHODS AND RESULTS: We studied the effect of PP1 on the anchorage-dependent growth of normal and v-Ha-Ras transformed NIH 3T3 fibroblasts, PAK activation and anchorage-independent growth of Ras transformants, and development of Ras-induced sarcomas in nude mice. We found that PP1 (10 nM) strongly inhibits PAK activity in Ras transformants. PP1 at this concentration is known to inhibit c-Fyn kinase, but not c-Src kinase, and none of the three known Src family kinases (c-Src, c-Fyn, and c-Yes) expressed in fibroblasts is activated by v-Ha-Ras. Thus, it is most likely that the primary target of this drug is an as yet unidentified Ras-activated Tyr (Y) kinase or kinases, which we call "Ray." Although PP1 has no effect on their anchorage dependent growth, it significantly inhibits their anchorage-independent growth in soft agar, as well as a rapid growth of Ras-induced sarcomas in mice. CONCLUSION: Like EGF receptor and ErbB2, a member of Src family kinases (most likely a new Src-related kinase called "Ray") is essential for the Ras-induced activation of PAK and the malignant transformation both in vitro and in vivo. These findings suggest that PP1 and other inhibitors specific for Src family kinases are potentially useful for the treatment of Ras-associated cancers. PMID- 11038145 TI - Intraoperative conformal optimization for transperineal prostate implantation using magnetic resonance spectroscopic imaging. AB - PURPOSE: Recent studies have demonstrated that magnetic resonance spectroscopic imaging (MRSI) of the prostate may effectively distinguish between regions of cancer and normal prostatic epithelium. This diagnostic imaging tool takes advantage of the increased choline and creatine versus citrate ratio found in malignant, compared with normal, prostate tissue. The purpose of this report is to present our initial experience integrating MRSI data into an intraoperative computer-based optimization planning system for prostate cancer patients who underwent permanent interstitial I 125 implantation. The goal of this approach was to achieve dose escalation to intraprostatic tumor deposits on the basis of MRSI findings without exceeding the tolerance of adjacent normal tissue structures. MATERIALS AND METHODS: MRSI was obtained before surgery for four consecutive patients with clinically localized prostate cancer. The ratios of choline and citrate for the prostate were analyzed, and regions in which malignant cells were suspected to be present were identified. These ratios were calculated on a spatial grid overlying the axial MRSI of the prostate. MRSI coordinates containing these suspicious regions were registered to the intraoperative ultrasound images. A computer-based treatment planning system, which relied on a genetic algorithm, was used to determine the optimal seed distribution necessary to achieve maximal target volume coverage with the prescription dose and to maintain urethra and rectal doses within tolerance ranges. The treatment planning system was specifically designed to escalate the dose to MRS-positive voxels while at the same time achieving preferential sparing of surrounding normal tissues. Patients underwent transperineal interstitial implantation with I 125 by use of this intraoperatively generated plan. Postimplant computed tomographic scans were performed on the same day of the procedure in all cases, and dosimetric guidelines of the American Brachytherapy Society were used to assess implant quality. RESULTS: Based on the postimplant computed tomographic evaluation, the intraoperative optimization treatment planning program was able to achieve a minimum dose of 139% to 192% of the 144-Gy prescription dose to the MRS-positive voxels. The percentage of the prostate volume receiving 100% of the prescription dose ranged from 92% to 97%, and the dose delivered to 90% of the target for the target volume ranged from 96% to 124%. Despite the dose escalation achieved for the positive voxels, the urethral and rectal doses were maintained within tolerance ranges. The average and maximal rectal doses ranged from 28% to 43% and 69% to 115% of the prescription dose, respectively. The average and maximal urethral doses ranged from 66% to 144% and 118% to 166% of the prescription dose, respectively. CONCLUSIONS: Using this brachytherapy optimization system, we could demonstrate the feasibility of MRS optimized dose distributions for I 125 permanent prostate implants. This approach may have an impact on the ability to select regions within the prostate to safely employ dose escalation for patients treated with permanent interstitial implantation and to improve outcome for patients with organ-confined prostatic cancers. PMID- 11038146 TI - Phase I and pharmacokinetic study of novobiocin in combination with VP-16 in patients with refractory malignancies. AB - PURPOSE: The coumarin antibiotic novobiocin potentiates the activity of etoposide (VP-16) in vitro by increasing intracellular accumulation of VP-16. The drug efflux pump inhibited by novobiocin appears to be distinct from both of the major proteins associated with the multidrug resistance phenotype in human cancers, the 170-kDa P-glycoprotein and the 190-kDa multidrug resistance protein. In a recent study, we found that novobiocin augmented VP-16 accumulation ex vivo in 16 of 24 fresh tumor samples at concentrations that could be achieved in vivo. Therefore, we conducted a clinical trial to determine the maximum tolerated dose and the pharmacokinetics of novobiocin when given in combination with VP-16. PATIENTS AND METHODS: Patients with refractory cancer were treated with VP-16 on days 1, 3, and 5. Antiemetics, consisting of ondansetron and dexamethasone, were given 60 minutes before the VP-16 was administered. Novobiocin was given orally 30 minutes before the VP-16, and the dose was escalated in successive groups of patients according to a standard dose escalation design. Treatment cycles were repeated every 4 weeks. Plasma concentrations of novobiocin were determined during the first treatment cycle by high-performance liquid chromatography. RESULTS: Thirty three patients were treated for a total of 69 cycles. Eleven patients were treated with a starting dose of VP-16 of 120 mg/m2, and three of these patients experienced neutropenic fever. The dose of VP-16 was reduced to 100 mg/m2, and an additional 22 patients were enrolled. The dose of novobiocin ranged from 3 to 9 g. At a novobiocin dose of at least 5.5 g, plasma concentrations of at least 150 microM were sustained for 24 hours. Dose-limiting toxicities consisted of neutropenic fever and reversible hyperbilirubinemia. Nausea, which was a limiting toxicity in other trials of novobiocin, was well controlled with the use of serotonergic antiemetics. Diarrhea was common but mild in most patients. DISCUSSION: In previously treated patients, the recommended dose of novobiocin in this schedule is 7 g/m2/day. Novobiocin does not appear to augment the toxicity of VP-16 to the bone marrow or the gastrointestinal mucosa. Plasma concentrations of novobiocin equivalent to the levels required to modulate VP-16 in vitro are readily achievable for total but not unbound free drug. PMID- 11038147 TI - The impact of contralateral breast cancer on the outcome of breast cancer patients treated by mastectomy. AB - PURPOSE: To evaluate the clinical and pathological features of breast cancer patients who develop contralateral breast cancer (CBC) and assess the impact of the second breast cancer on their prognosis. PATIENTS AND METHODS: This retrospective study includes 2136 women with stage I-III breast cancer treated between 1927 and 1987 at the University of Chicago Hospitals. A total of 132 (6.2%) developed CBC during a median follow-up period of 14.2 years; all of them were treated with mastectomy for both breast cancers. We compare the prognostic characteristics, treatments, and outcomes of patients who developed bilateral breast cancer with those who had only unilateral breast cancer (UBC). We also compare the features of the first and the second tumors among patients with bilateral breast cancer (BBC). RESULTS: The annual incidence rate for CBC remained constant at an average rate of 0.23%, resulting in a cumulative incidence rate of 6.2%. Patients with BBC were significantly younger than those with UBC (median age, 51 years vs 54 years). No other significant differences were observed between BBC and UBC patients. Among BBC patients, the second cancer was smaller (2.0 cm vs 3.0 cm) and was associated with a lower incidence of axillary node involvement (29% vs 52%). The development of CBC was associated with worse survival (hazard ratio = 1.46 in comparison with patients who did not develop CBC, 95% CI of 1.09-1.95). On multivariate analysis, factors that decreased the disease-specific survival (DSS) in patients with BBC were a higher number of positive lymph nodes of the first and second cancers, a larger size of the second cancer, and a shorter interval between the two primaries. DISCUSSION: At the time of diagnosis of first breast cancer, BBC patients were significantly younger than UBC patients. The second cancer among the BBC patients was at an earlier stage than the first one; however, no difference was noticed in the pathological feature between the cancer in the UBC patients and the first cancer of BBC patients. There is an indication that the longer the interval between the two cancers, the better the survival of the BBC patients. PMID- 11038148 TI - Two randomised and placebo-controlled studies of an oral prostacyclin analogue (Iloprost) in severe leg ischaemia. The Oral Iloprost in severe Leg Ischaemia Study Group. AB - Two separate studies are described using the same prostacyclin analogue in a similar group of patients. OBJECTIVES: to assess the tolerability and efficacy of two dose regimens of oral Iloprost compared with placebo in the treatment of patients with ischaemic ulcers, gangrene or rest pain due to severe arterial disease over a period of 4 weeks (Study A) and one year (Study B). DESIGN: multicentre, placebo controlled, double-blind, randomized prospective studies. SUBJECTS & METHODS: 178 (study A) and 624 (study B) patients with trophic skin lesions (ulcers or gangrene) or ischaemic rest pain due to severe arterial disease. To confirm severe arterial disease patients were required to have a systolic ankle Doppler pressure of 70 mmHg or less or a toe systolic Doppler pressure of 50 mmHg or less in one leg. In both studies patients were randomly allocated to three treatment groups: placebo, low dose Iloprost (50-100 microgram twice a day) or high dose (150-200 microgram twice a day) In Study A the main outcome measures were tolerability of different doses of Iloprost and death, major amputation, healing of trophic lesions and relief of rest pain at the end of the follow up, which was 5 months after the end of the treatment. In Study B the primary end point was time to major amputation and stroke or death up to 12 months. Secondary pre-defined end points included the combined end point of patients alive without amputation, no trophic skin changes, no rest pain and not on regular analgesics. RESULTS: the proportion of patients who completed the 4 week treatment period in Study A at the intended dose was 58%, 43%, 45% respectively in the placebo, low dose and high dose Iloprost groups. In an intention to treat analysis the proportion of patients who survived without major amputation, ulcers or gangrene and had no rest pain was 11% in the placebo group, 19% in the low dose iloprost group and 28% in the high dose Iloprost group. The pooled Iloprost groups showed a statistically significantly better result than the placebo group (p=0.04), as did the high dose Iloprost group compared to the placebo (p=0.014). In Study B there was no treatment benefit in terms of a primary end point of amputation and death. However the secondary combined end point of patients who survived without a major amputation, ulcers or gangrene and had no rest pain, nor a need for regular analgesia was favourable for Iloprost, with 18% of patients in the placebo group reaching this optimal secondary end point, compared to 23% in the low dose Iloprost group and 26% in the higher dose Iloprost group (p<0.05). CONCLUSIONS: oral Iloprost administered for a year showed no clear benefit in patients with advanced severe leg ischaemia (PAOD III and IV). The results obtained with 4 weeks' treatment in Study A and in previous trials of intravenous Iloprost could not be reproduced PMID- 11038149 TI - Specificity of cytochrome P450 2A3-catalyzed alpha-hydroxylation of N' nitrosonornicotine enantiomers. AB - N'-nitrosonornicotine (NNN) induces tumors in the rat nasal cavity and esophagus and is believed to be a causative agent for esophageal cancer in tobacco users. To exert its carcinogenic potential, NNN must be metabolically activated by alpha hydroxylation at either the 2'- or 5'-carbon. We previously reported that the human cytochrome P450 (P450), 2A6, efficiently and specifically catalyzed NNN 5' hydroxylation. P450 2A3, which is expressed in the rat nasal cavity and to a small extent in the esophagus, is closely related to P450 2A6. P450 2A3, like 2A6, is a good catalyst of NNN alpha-hydroxylation (K(m) 7 microM; V(max) 17 nmol/min/nmol). However, in contrast to P450 2A6, 2A3 catalyzed both 5'- and 2' hydroxylation of NNN. The ratio of 2'- to 5'-hydroxylation was 1:3. These data, both with P450 2A6 and 2A3, were obtained using racemic NNN. P450 2A3 catalyzed metabolism of (S)-NNN occurred exclusively at the 5'-position. The predominant pathway of (R)-NNN metabolism was 2'-hydroxylation, and occurred to a 3-fold greater extent than did 5'-hydroxylation. These data are in contrast to those obtained from a recent study of (R)- and (S)-NNN metabolism by cultured rat esophagus. In that study, (S)-NNN was metabolized predominantly by 2' hydroxylation and (R)-NNN equally by 2'- and 5'-hydroxylation. Taken together, these data provide strong evidence that P450 2A3 is not the rat esophageal P450 that catalyzes the metabolic activation of NNN. P450 2A3 may be an important catalyst of NNN activation in rat nasal mucosa. PMID- 11038150 TI - Chronic nifedipine dosing enhances cephalexin bioavailability and intestinal absorption in conscious rats. AB - Cephalexin, a beta-lactam antibiotic, is rapidly absorbed via the di-and tripeptide intestinal transporters, as for many peptidomimetic drugs. Acute nifedipine has been shown to increase intestinal absorption of several beta lactams: amoxicillin and cefixime in humans, and cephalexin in the rat. We showed previously that the nervous system was involved in the increasing effect of nifedipine on cephalexin intestinal absorption in anesthetized rats. The aim of the present study was 2-fold: 1) to investigate whether the effect of nifedipine is maintained in conscious rats, and 2) to determine whether the nifedipine effect will persist during chronic nifedipine administration. Acute and chronic oral administration of nifedipine significantly increased oral cephalexin area under the plasma concentration-time curve (34 and 25%, respectively) and maximum concentration in plasma (57 and 51%, respectively), while the distribution and elimination parameters of intra-arterial cephalexin were not affected by acute or chronic nifedipine administration. In conclusion, acute nifedipine effect on intestinal absorption of cephalexin is independent of anesthesia in rats. Since nifedipine could still enhance cephalexin intestinal absorption after a 7-day b.i.d. treatment, it can be envisaged to apply this effect to increase bioavailability of poorly absorbed peptidomimetic drugs in man. PMID- 11038151 TI - Milk thistle, a herbal supplement, decreases the activity of CYP3A4 and uridine diphosphoglucuronosyl transferase in human hepatocyte cultures. AB - Milk thistle extract is one of the most commonly used nontraditional therapies, particularly in Germany. Milk thistle is known to contain a number of flavonolignans. We evaluated the effect of silymarin, on the activity of various hepatic drug-metabolizing enzymes in human hepatocyte cultures. Treatment with silymarin (0.1 and 0.25 mM) significantly reduced the activity of CYP3A4 enzyme (by 50 and 100%, respectively) as determined by the formation of 6-beta-hydroxy testosterone and the activity of uridine diphosphoglucuronosyl transferase (UGT1A6/9) (by 65 and 100%, respectively) as measured by the formation of 4 methylumbelliferone glucuronide. Silymarin (0.5 mM) also significantly decreased mitochondrial respiration as determined by MTT reduction in human hepatocytes. These observations point to the potential of silymarin to impair hepatic metabolism of certain coadministered drugs in humans. Indiscriminate use of herbal products may lead to altered pharmacokinetics of certain drugs and may result in increased toxicity of certain drugs. PMID- 11038152 TI - Metabolites of caspofungin acetate, a potent antifungal agent, in human plasma and urine. AB - Caspofungin acetate (MK-0991) is a semisynthetic pneumocandin derivative being developed as a parenteral antifungal agent with broad-spectrum activity against systemic infections such as those caused by Candida and Aspergillus species. Following a 1-h i.v. infusion of 70 mg of [(3)H]MK-0991 to healthy subjects, excretion of drug-related material was very slow, such that 41 and 35% of the dosed radioactivity was recovered in urine and feces, respectively, over 27 days. Plasma and urine samples collected around 24 h postdose contained predominantly unchanged MK-0991, together with trace amounts of a peptide hydrolysis product, M0, a linear peptide. However, at later sampling times, M0 proved to be the major circulating component, whereas corresponding urine specimens contained mainly the hydrolytic metabolites M1 and M2, together with M0 and unchanged MK-0991, whose cumulative urinary excretion over the first 16 days postdose represented 13, 71, 1, and 9%, respectively, of the urinary radioactivity. The major metabolite, M2, was highly polar and extremely unstable under acidic conditions when it was converted to a less polar product identified as N-acetyl-4(S)-hydroxy-4-(4 hydroxyphenyl)-L-threonine gamma-lactone. Derivatization of M2 in aqueous media led to its identification as the corresponding gamma-hydroxy acid, N-acetyl-4(S) hydroxy-4-(4-hydroxyphenyl)-L-threonine. Metabolite M1, which was extremely polar, eluting from HPLC column just after the void volume, was identified by chemical derivatization as des-acetyl-M2. Thus, the major urinary and plasma metabolites of MK-0991 resulted from peptide hydrolysis and/or N-acetylation. PMID- 11038153 TI - Heterologous expression of CYP2K1 and identification of the expressed protein (BV CYP2K1) as lauric acid (omega-1)-hydroxylase and aflatoxin B1 exo-epoxidase. AB - LMC2 is the most abundant constitutively expressed hepatic cytochrome P450 found in sexually immature rainbow trout (Onchorynchus mykiss) and is also the isozyme that activates the carcinogen aflatoxin B1 (AFB1). This P450 has been cloned, sequenced, and designated as CYP2K1. The present report describes the heterologous expression of enzymatically active CYP2K1 (BV-CYP2K1) in baculovirus Spodoptera frugiperda (Sf9) insect cells and its catalytic and immunoreactivity characterization in comparison with that of the previously purified LMC2 P450. Homogenates of Sf9 cells expressing the CYP2K1 enzyme and LMC2 both catalyzed the hydroxylation of lauric acid and the epoxidation of AFB1 in the presence of rat NADPH-cytochrome P450 reductase. Both LMC2 and BV-CYP2K1 catalyzed the oxidation of lauric acid primarily at the (omega-1) position plus small amounts at the (omega-2) position. Formation of AFB1 epoxide was shown indirectly by the appearance of an AFB1 epoxide-glutathione conjugate when P450 incubation mixtures contained AFB1, glutathione (GSH) together with mouse liver cytosol or purified rat GSH-transferase. When the AFB1 epoxide-GSH conjugate produced by BV-CYP2K1 and purified LMC2 was analyzed by HPLC using a chiral column, it had a retention time identical to that produced by CYP3A4, a human P450 known to form exclusively the AFB1 exo-epoxide. These results, therefore, confirm that the cDNA-expressed CYP2K1 protein is catalytically and immunologically identical to purified trout LMC2 and that these two enzymes produce primarily the highly carcinogenic stereoisomeric exo-epoxide form of AFB1. PMID- 11038154 TI - Cytochrome P4502C9 is the principal catalyst of racemic acenocoumarol hydroxylation reactions in human liver microsomes. AB - The oral anticoagulant acenocoumarol is given as a racemic mixture. The (S) enantiomer is rapidly cleared and is the reason why only (R)-acenocoumarol contributes to the pharmacological effect. The objective of the study was to establish the cytochrome P450 (CYP) enzymes catalyzing the hydroxylations of the acenocoumarol enantiomers. Of various cDNA-expressed human CYPs, only CYP2C9 hydroxylated (S)-acenocoumarol. Hydroxylation occurred at the 6-, 7-, and 8 position with equal K(m) values and a ratio of 0.9:1:0.1 for V(max). CYP2C9 also mediated the 6-, 7-, and 8-hydroxylations of (R)-acenocoumarol with K(m) values three to four times and V(max) values one-sixth times those of (S)-acenocoumarol. (R)-Acenocoumarol was also metabolized by CYP1A2 (6-hydroxylation) and CYP2C19 (6 , 7-, and 8-hydroxylation). In human liver microsomes one enzyme only catalyzed (S)-acenocoumarol hydroxylations with K(m) values < 1 microM. In most of the samples tested the 7-hydroxylation of (R)-acenocoumarol was also catalyzed by one enzyme only. The 6-hydroxylation was catalyzed by at least two enzymes. Sulfaphenazole could completely inhibit in a competitive way the hydroxylations of (S)-acenocoumarol and the 7-hydroxylation of (R)-acenocoumarol. The 6 hydroxylation of (R)-acenocoumarol could be partially inhibited by sulfaphenazole, 40 to 50%, and by furafylline, 20 to 30%. Significant mutual correlations were obtained between the hydroxylations of (S)-acenocoumarol, the 7 hydroxylation of (R)-acenocoumarol, the 7-hydroxylation of (S)-warfarin, and the methylhydroxylation of tolbutamide. The results demonstrate that (S) acenocoumarol is hydroxylated by a single enzyme, namely CYP2C9. CYP2C9 is also the main enzyme in the 7-hydroxylation of (R)-acenocoumarol. Other enzymes involved in (R)-acenocoumarol hydroxylation reactions are CYP1A2 and CYP2C19. Drug interactions must be expected, particularly for drugs interfering with CYP2C9. Also, drugs interfering with CYP1A2 and CYP2C19 may potentiate acenocoumarol anticoagulant therapy. PMID- 11038155 TI - CYP3A4 is a major isoform responsible for oxidation of 7-hydroxy-Delta(8) tetrahydrocannabinol to 7-oxo-delta(8)-tetrahydrocannabinol in human liver microsomes. AB - The human liver enzyme microsomal alcohol oxygenase was able to oxidize both 7alpha- and 7beta-hydroxy-Delta(8)-tetrahydrocannabinol (7alpha- and 7beta hydroxy-Delta(8)-THC) to 7-oxo-Delta(8)-THC. The oxidative activity was determined by using a panel of 12 individual cDNA-expressed human cytochrome P450s (CYPs) (1A1, 1A2, 2A6, 2B6, 2C8, 2C9-Arg, 2C9-Cys, 2C19, 2D6-Met, 2D6-Val, 2E1 and 3A4). Among the CYP isoforms examined, CYP3A4 showed the highest activity for both of substrates. The metabolism of 7alpha- and 7beta-hydroxy-Delta(8)-THC to 7-oxo-Delta(8)-THC was also detected for CYPs 1A1 (4.8% of CYP3A4), 1A2 (4.7%), 2A6 (2.3%), 2C8 (16.6%), and 2C9-Cys (5.4%), and CYPs 1A1 (0.4%), 2C8 (1.3%), 2C9-Arg (4.3%), and 2C9-Cys (0.9%), respectively. The 7alpha- and 7beta hydroxy-Delta(8)-THC microsomal alcohol oxygenase activities in human liver were significantly inhibited by addition of 100 microM troleandomycin, 1 microM ketoconazole, and anti-CYP3A antibody, although these activities were not inhibited by 1 microM 7, 8-benzoflavone and 50 microM sulfaphenazole. When the substrates were incubated with the CYP3A4-expressed microsomes under oxygen-18 gas phase, atmospheric oxygen was incorporated into 35% of 7-oxo-Delta(8)-THC formed from 7alpha-OH-Delta(8)-THC, but only 12% of 7-oxo-Delta(8)-THC formed from 7beta-OH-Delta(8)-THC. These results indicate that CYP3A4 is a major isoform responsible for the oxidation of 7alpha- and 7beta-hydroxy-Delta(8)-THC to 7-oxo Delta(8)-THC in liver microsomes of humans, although the oxidation mechanisms for 7alpha- and 7beta-hydroxy-Delta(8)-THC might be different. PMID- 11038156 TI - Prenylflavonoids from hops inhibit the metabolic activation of the carcinogenic heterocyclic amine 2-amino-3-methylimidazo[4, 5-f]quinoline, mediated by cDNA expressed human CYP1A2. AB - The heterocyclic amine 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) is a potential human carcinogen found in cooked food that requires initial metabolic activation by cytochrome P450s, primarily CYP1A2. The present study was conducted to examine whether recombinant human CYP1A2 expressed in insect cells mediates the metabolic activation of IQ and whether prenylflavonoids found in hops and beer would modulate the CYP1A2-mediated activation of IQ. The cDNA-expressed human CYP1A2 was found to strongly activate IQ as measured by the Ames Salmonella assay and by the covalent binding of IQ metabolites to calf thymus DNA and protein. Inhibition studies showed that the prenylchalcone xanthohumol and the prenylflavanones 8-prenylnaringenin and isoxanthohumol strongly inhibited the mutagenic activation of IQ mediated by cDNA-expressed human CYP1A2 in the Ames Salmonella assay. The three prenylflavonoids also markedly inhibited the human CYP1A2-mediated binding of IQ to metabolites that bind to DNA. The inhibition of the metabolic activation of IQ was paralleled by the inhibition of acetanilide 4 hydroxylase activity of human CYP1A2. Thus, xanthohumol, isoxanthohumol, and prenylflavanones 8-prenylnaringenin are potent inhibitors of the metabolic activation of IQ and may have the potential to act as chemopreventive agents against cancer induced by heterocyclic amines activated by CYP1A2. PMID- 11038157 TI - A significant role of human cytochrome P450 2C8 in amiodarone N-deethylation: an approach to predict the contribution with relative activity factor. AB - Human cytochrome P450 (CYP) isoforms involved in amiodarone N-deethylation were identified, and the relative contributions of these CYP isoforms were evaluated in different human liver microsomes. The mean K(M) and V(max) values of amiodarone N-deethylation in microsomes from six human livers were 31.6 +/- 7.5 microM and 1.2 +/- 0.7 pmol/min/pmol of CYP, respectively. Ketoconazole and anti CYP3A antibodies strongly inhibited amiodarone N-deethylase activity in human liver microsomes at a substrate concentration of 50 microM. Of 15 recombinant human CYP enzymes (19 preparations), CYP1A1, CYP3A4, CYP1A2, CYP2D6, CYP2C8, and CYP2C19 catalyzed amiodarone N-deethylation. The amiodarone N-deethylase activity at a substrate concentration of 5 microM was significantly correlated with the paclitaxel 6alpha-hydroxylase activity (r = 0.84, P <.05) in the human liver microsomes, whereas the amiodarone N-deethylase activity at 100 microM was significantly correlated with the testosterone 6beta-hydroxylase activity (r = 0.94, P <.005). According to the concept of relative activity factor, it was clarified that CYP2C8 as well as CYP3A4 were significantly involved in amiodarone N-deethylation in human livers at clinically significant concentrations and that the contributions of CYP1A2, CYP2C19, and CYP2D6 were relatively minor. However, there was a large interindividual variability in the contribution of each CYP isoform to amiodarone N-deethylase activity in human liver; the relevance of these enzymes would be dependent on the content of the respective isoforms and on the amiodarone concentration in the liver. PMID- 11038158 TI - Induction of mouse CYP2J by pyrazole in the eye, kidney, liver, lung, olfactory mucosa, and small intestine, but not in the heart. AB - We have recently shown that rat CYP2J4 is inducible by pyrazole in liver, small intestine, and olfactory mucosa. The aim of the present study was to determine whether mouse CYP2Js are also inducible by pyrazole, which was known to induce CYP2A5 in mouse liver and kidney, but not in lung or olfactory mucosa. CYP2J proteins were detected in mouse liver, lung, kidney, heart, eye, olfactory mucosa, and small intestine by immunoblot analysis with an anti-CYP2J4 antibody. The microsomal level of the CYP2J4-related P450s in various mouse tissues ranked in the order of small intestine > olfactory mucosa > liver > kidney > or = heart > lung > eye. Induction of the CYP2J proteins was observed in the eye, liver, lung, kidney, olfactory mucosa, and small intestine, but not in the heart, after daily i.p. injection of pyrazole at 120 or 200 mg/kg for 3 days. CYP2J proteins were induced similarly in C57BL/6 and DBA/2 mice. CYP2A5 was detected in the small intestine in addition to liver and olfactory mucosa; however, treatment with pyrazole induced CYP2A5 in the liver, but not in the olfactory mucosa or the small intestine. Induction of CYP2J mRNAs was also observed by RNA blot analysis with a CYP2J4 cDNA probe. RNA-polymerase chain reaction analysis showed that, in both untreated and pyrazole-treated mice, CYP2J5 was expressed in the kidney and liver, but not in the other tissues examined, whereas CYP2J6 was detected in all tissues examined. The different tissue selectivities in CYP2A5 and CYP2J induction by pyrazole suggest involvement of different regulatory mechanisms. PMID- 11038159 TI - Decreased in vivo metabolism of drugs in chronic renal failure. AB - Chronic renal failure (CRF) is associated with a decrease in renal excretion of drugs, but its effects on the liver metabolism of xenobiotics are poorly defined. The objectives of this study were to determine the effects of CRF on hepatic cytochrome P450 (CYP450) and its repercussions on in vivo hepatic metabolism of drugs. Two groups of rats were studied: control paired-fed and CRF. CRF was induced by subtotal nephrectomy. Total CYP450 activity and protein expression of several CYP450 isoforms (CYP1A2, CYP2C11, CYP3A1, CYP3A2) were assessed in liver microsomes. In vivo cytochrome P450 activity was evaluated with breath tests using substrates for different isoenzymes: caffeine (CYP1A2), aminopyrine (CYP2C11), and erythromycin (CYP3A2). Creatinine clearance was reduced by 60% (P <. 01) in rats with CRF. Compared with control paired-fed rats, total CYP450 activity was reduced by 40% in rats with CRF. Protein expression of CYP2C11, CYP3A1, and CYP3A2 was considerably reduced (more than 45%, P <.001) in rats with CRF, whereas the levels of CYP1A2 were unchanged. In rats with CRF, there was a 35% reduction in the aminopyrine (CYP2C11) and the erythromycin (CYP3A2) breath tests compared with control animals (P <.001). The caffeine (CYP1A2) breath tests remained comparable to controls. Creatinine clearance correlated with the aminopyrine and erythromycin breath tests (r(2) = 0.73 and r(2) = 0.81, respectively, P <.001). In conclusion, CRF is associated with a decrease in total liver CYP450 activity in rats (mainly in CYP2C11, CYP3A1, and CYP3A2), which leads to a significant decrease in the metabolism of drugs. PMID- 11038160 TI - CYP2A5/CYP2A6 expression in mouse and human hepatocytes treated with various in vivo inducers. AB - Induction of coumarin 7-hydroxylation, catalyzed by CYP2A5 in mice and CYP2A6 in humans by various known in vivo murine inducers and modifiers, was compared in human and mouse hepatocytes in culture. Phenobarbital and rifampicin were efficient inducers (up to 10-fold induction) after 48-h treatment in murine cultured hepatocytes, whereas the enzyme activity in human hepatocytes was much more refractory to induction. However, a prolongation of incubation time to 72 h in human hepatocytes led to a modest restoration of inducibility by phenobarbital. Of the three porphyrinogenic inducers studied, griseofulvin induced the murine enzyme efficiently, but not the human enzyme, whereas aminotriazole and thioacetamide had no effect on either species. Pyrazole produced substantial induction in both human and murine hepatocytes, whereas cobalt chloride, which is also an in vivo inducer of the mouse enzyme, had no effect. Clofibric acid, an in vivo depressor of coumarin 7-hydroxylase, also depressed hepatocyte activities. In both murine and human hepatocytes, changes in CYP2A5/6 mRNA levels correlated roughly with enzyme changes, except in the case of cobalt chloride, which increased mRNA levels despite a lack of effect on enzyme activity. In general, human and mouse hepatocytes gave a similar response to CYP2A inducers. However, some differences were found, which means that, although CYP2A isozymes are probably regulated in a similar manner in both species, it is necessary to be cautious before extrapolating to human the results found in mouse models. PMID- 11038161 TI - Automated definition of the enzymology of drug oxidation by the major human drug metabolizing cytochrome P450s. AB - A fully automated assay to determine the enzymology of drug oxidation by the major human hepatic cytochrome P450s (CYPs; CYP1A2, -2C9, -2C19, -2D6, and -3A4) coexpressed functionally in Escherichia coli with human NADPH-P450 reductase has been developed and validated. Ten prototypic substrates were chosen for which clearance was primarily CYP-dependent, and the activities of these five major CYPs were represented. A range of intrinsic clearance (CL(int)) values were obtained for substrates in both pooled human liver microsomes (HLM; 1-380 microl. min(-1)mg(-1)) and recombinant CYPs (0.03-7 microl. min(-1)pmol(-1)) and thus the percentage contribution of individual CYPs toward their oxidative metabolism could be estimated. All the assignments were consistent with the available literature data. Tolbutamide was metabolized by CYP2C9 (70%) and CYP2C19 (30%), diazepam by CYP2C19 (100%), ibuprofen by CYP2C9 (90%) and CYP2C19 (10%), and omeprazole by CYP2C19 (68%) and CYP3A4 (32%). Metoprolol and dextromethorphan were primarily CYP2D6 substrates and propranolol was metabolized by CYP2D6 (59%), CYP1A2 (26%), and CYP2C19 (15%). Diltiazem, testosterone, and verapamil were metabolized predominantly by CYP3A4. In addition, the metabolite profile for the CYP-dependent clearance of several markers determined by mass spectroscopy was as predicted from the literature. There was a good correlation between the sum of individual CYP CL(int) and HLM CL(int) (r(2) = 0.8, P <.001) for the substrates indicating that recombinant CYPs may be used to predict HLM CL(int) data. This report demonstrates that recombinant human CYPs may be useful as an approach for the prediction of the enzymology of human CYP metabolism early in the drug discovery process. PMID- 11038162 TI - Human serum paraoxonase (PON1) isozymes Q and R hydrolyze lactones and cyclic carbonate esters. AB - It is well established that human serum paraoxonase (PON1) catalyzes the hydrolysis of organophosphate insecticides and nerve agents, as well as that of a number of aromatic carboxylic acid esters. Our laboratory has recently found a new class of PON1 substrates that includes at least 30 lactones and cyclic carbonate esters. The lactone substrates vary in their ring size from 4 to 7 atoms. Substituents on the ring carbons may enhance or reduce the rate of lactone hydrolysis. An appreciable degree of stereospecificity exists with some activities differing up to 9-fold between enantiomers (i.e., S-alpha-hydroxy gamma-butyrolactone is hydrolyzed 5 to 9 times faster than the R form). Thiolactones are hydrolyzed less efficiently, and some lactams are potent inhibitors. Four lactone-containing drugs-spironolactone, mevastatin, simvastatin, and lovastatin-have been identified as substrates for PON1. All lactone substrates are hydrolyzed by both the Q and R isozymes of human serum PON1. However, some lactone substrates are hydrolyzed faster by the Q than R isozyme, whereas others show a reverse preference. Moreover, these new substrates include homogentisic acid lactone, mevalonic acid lactone, homocysteine thiolactone, and gamma-hydroxybutyric acid lactone-all lactone forms of endogenous compounds. It is reasonable to expect that further investigations may uncover PON1 lactone substrates that are, themselves, endogenous compounds. In this article we characterize the basic enzymatic properties of PON1's newly identified hydrolytic activities with lactone and cyclic carbonate ester substrates and compare these properties with those of representative arylesters and organophosphates. PMID- 11038163 TI - In vitro evaluation of the disposition of A novel cysteine protease inhibitor. AB - K11777 (N-methyl-piperazine-Phe-homoPhe-vinylsulfone-phenyl) is a potent, irreversible cysteine protease inhibitor. Its therapeutic targets are cruzain, a cysteine protease of the protozoan parasite Trypanosoma cruzi, and cathepsins B and L, which are associated with cancer progression. We evaluated the metabolism of K11777 by human liver microsomes, isolated cytochrome P450 (CYP) enzymes, and flavin-containing monooxygenase 3 (FMO3) in vitro. K11777 was metabolized by human liver microsomes to three major metabolites: N-oxide K11777 (apparent K(m) = 14.0 +/- 4.5 microM and apparent V(max) = 3460 +/- 3190 pmol. mg(-1). min(-1), n = 4), beta-hydroxy-homoPhe K11777 (K(m) = 16.8 +/- 3.5 microM and V(max) = 1260 +/- 1090 pmol. mg(-1). min(-1), n = 4), and N-desmethyl K11777 (K(m) = 18.3 +/- 7.0 microM and V(max) = 2070 +/- 1830 pmol. mg(-1). min(-1), n = 4). All three K11777 metabolites were formed by isolated CYP3A and their formation by human liver microsomes was inhibited by the CYP3A inhibitor cyclosporine (50 microM, 54 62% inhibition) and antibodies against human CYP3A4/5 (100 microg of antibodies/100 microg microsomal protein, 55-68% inhibition). CYP2D6 metabolized K11777 to its N-desmethyl metabolite with an apparent K(m) (9.2 +/- 1.4 microM) lower than for CYP3A4 (25.0 +/- 4.0 microM) and human liver microsomes. The apparent K(m) for N-oxide K11777 formation by cDNA-expressed FMO3 was 109 +/- 11 microM. Based on the intrinsic formation clearances and the results of inhibition experiments (CYP2D6, 50 microM bufuralol; FMO3 mediated, 100 mM methionine) using human liver microsomes, it was estimated that CYP3A contributes to >80% of K11777 metabolite formation. K11777 was a potent (IC(50) = 0.06 microM) and efficacious (maximum inhibition 85%) NADPH-dependent inhibitor of human CYP3A4 mediated 6'beta-hydroxy lovastatin formation, suggesting that K11777 is not only a substrate but also a mechanism-based inhibitor of CYP3A4. PMID- 11038164 TI - O-Glucuronidation of the lung carcinogen 4-(methylnitrosamino)-1- (3-pyridyl)-1 butanol (NNAL) by human UDP-glucuronosyltransferases 2B7 and 1A9. AB - 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and its major metabolite, 4 (methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), are potent lung carcinogens in animals. UGT-mediated O-glucuronidation of NNAL is an important detoxification pathway for these carcinogens. To better characterize this pathway in humans, we screened a series of UGT-overexpressing cell lines and baculosome preparations for their ability to O-glucuronidate NNAL and examined multiple human liver and lung specimens for NNAL-glucuronidating activity and their levels of expression of NNAL-glucuronidating UGTs. Human liver microsomal fractions exhibited significant levels of NNAL-glucuronidating activity, with the NNAL-Gluc II diastereomer formed at a rate 3.4 times that observed for NNAL-Gluc I. As with liver microsomal fractions, NNAL-Gluc II was the major diastereomer formed by homogenates from UGT2B7-overexpressing HK293 cells or UGT2B7-overexpressing baculosomes; the major diastereomer formed by homogenates from UGT1A9 overexpressing V79 cells was NNAL-Gluc I. No significant O-glucuronidating activity of NNAL was detected in UGT1A1-, UGT1A4-, UGT1A6-, UGT2B4-, or UGT2B15 overexpressing HK293 or V79 cell homogenates, or in UGT1A1-, UGT1A3-, UGT1A7-, or UGT1A10-overexpressing baculosomes. Significant levels of UGT2B7 mRNA were detected by reverse transcriptase-polymerase chain reaction in human liver and at low levels in human lung specimens. UGT1A9 mRNA was detected in liver but not in lung. These results suggest that although both UGT2B7 and UGT1A9 play an important role in the overall glucuronidation of NNAL in humans, UGT2B7 potentially plays an important role in the detoxification of NNAL in the lung. PMID- 11038165 TI - Formation of a dihydroxy metabolite of phenytoin in human liver microsomes/cytosol: roles of cytochromes P450 2C9, 2C19, and 3A4. AB - Formation of four oxidative metabolites from the anticonvulsant drug phenytoin (DPH) catalyzed by human liver microsomal cytochrome P450 (P450) enzymes was determined simultaneously. Under the conditions in which linearity for formation of 4'-hydroxylated DPH (4'-HPPH; main metabolite) was observed, human liver cytosol increased microsome-mediated DPH oxidation. 3',4'-Dihydroxylated product (3', 4'-diHPPH) formation was 10 to 40% of total DPH oxidation in the presence of liver cytosol. 3'-Hydroxy DPH formation was catalyzed by only one of the human liver microsomal samples examined and 3', 4'-dihydrodiol formation could not be detected in all samples. In the presence of liver cytosol, 3',4'-diHPPH formation activity from 100 microM 4'-HPPH was correlated with testosterone 6beta hydroxylation activity and CYP3A4 content. However, 3', 4'-diHPPH formation using 1 or 10 microM 4'-HPPH as a substrate was not correlated with contents of any P450s or marker activities. Of 10 cDNA-expressed human P450 enzymes examined, CYP2C19, CYP2C9, and CYP3A4 catalyzed 3',4'-diHPPH formation from the primary hydroxylated metabolites (3'-hydroxy-DPH and 4'-HPPH). Fluvoxamine and anti-CYP2C antibody inhibited 3',4'-diHPPH formation from 10 microM 4'-HPPH in a human liver sample that contained relatively high levels of CYP2C, whereas ketoconazole and anti-CYP3A antibody showed inhibitory effects on the activities in liver microsomal samples in which CYP3A4 levels were relatively high. These results suggest that CYP2C9, CYP2C19, and CYP3A4 all have catalytic activities in 3',4' diHPPH formation from primary hydroxylated metabolites in human liver and that the hepatic contents of these three P450 forms determine which P450 enzymes play major roles of DPH oxidation in individual humans. PMID- 11038166 TI - Lactonization is the critical first step in the disposition of the 3-hydroxy-3 methylglutaryl-CoA reductase inhibitor atorvastatin. AB - In an in vitro study, we compared the cytochrome P450 (CYP)-dependent metabolism and drug interactions of the acid and lactone forms of the 3-hydroxy-3 methylglutaryl (HMG)-CoA reductase inhibitor atorvastatin. Metabolism of atorvastatin acid and lactone by human liver microsomes resulted in para-hydroxy and ortho-hydroxy metabolites. Both substrates were metabolized mainly by CYP3A4 and CYP3A5. Atorvastatin lactone had a significantly higher affinity to CYP3A4 than the acid (K(m): para-hydroxy atorvastatin, 25.6 +/- 5.0 microM; para-hydroxy atorvastatin lactone, 1.4 +/- 0.2 microM; ortho-hydroxy atorvastatin, 29.7 +/- 9.4 microM; and ortho-hydroxy atorvastatin lactone, 3.9 +/- 0.2 microM). Compared with atorvastatin acid, CYP-dependent metabolism of atorvastatin lactone to its para-hydroxy metabolite was 83-fold higher [formation CL(int) (V(max)/K(m)): lactone 2949 +/- 3511 versus acid 35.5 +/- 48.1 microl. min(-1). mg(-1)] and to its ortho-hydroxy metabolite was 20-fold higher (CL(int): lactone 923 +/- 965 versus acid 45.8 +/- 59. 1 microl. min(-1). mg(-1)). Atorvastatin lactone inhibited the metabolism of atorvastatin acid by human liver microsomes with an inhibition constant (K(i)) of 0.9 microM while the K(i) for inhibition of atorvastatin by atorvastatin lactone was 90 microM. Binding free energy calculations of atorvastatin acid and atorvastatin lactone complexed with CYP3A4 revealed that the smaller desolvation energy of the neutral lactone compared with the anionic acid is the dominant contribution to the higher binding affinity of the lactone rather than an entropy advantage. Because atorvastatin lactone has a significantly higher metabolic clearance and the lactone is a strong inhibitor of atorvastatin acid metabolism, it can be expected that metabolism of the lactone is the relevant pathway for atorvastatin elimination and drug interactions. We hypothesize that most of the open acid metabolites present in human plasma are generated by interconversion of lactone metabolites. PMID- 11038167 TI - Early biotransformations of oxaliplatin after its intravenous administration to cancer patients. AB - This article deals with the fate of oxaliplatin 1 and 3 h after its i.v. administration (130 mg/m(2)) to three patients. Its binding to plasma proteins and penetration into red blood cells were monitored by chromatography on-line with inductively coupled plasma mass spectrometry. Oxaliplatin biotransformations in plasma ultrafiltrate (PUF) and in urine were studied by chromatography coupled to inductively coupled plasma mass spectrometry or to electrospray ionization mass spectrometry. In plasma, four platinum (Pt) compounds were found. The peaks at 200 and 160 kDa corresponding to gamma-globulins contained 40% of the Pt bound; the peak at 60 kDa corresponding to albumin contained 40% of the Pt found. The peak <2 kDa could correspond to oxaliplatin, to its degradation products, or to adducts between Pt compounds and low-molecular-weight species such as glutathione, L-methionine, and L-cysteine. In PUF and urine, oxaliplatin itself, its degradation products, Pt(dach)Cl(2), [Pt(dach)(OH(2))Cl](+), and species that have the same retention times as Pt(dach)(methionine) and [Pt(dach)](2)(glutathione) were found. One hour after infusion, oxaliplatin in PUF and urine represented 12 and 50% of the total Pt, respectively. Three hours after infusion, oxaliplatin, undetectable in PUF, represented 10% of total Pt in urine. Inside red blood cells, two Pt compounds were found. The Pt peak at 60 kDa corresponding to hemoglobin and the peak <2 kDa corresponding to low-molecular species contained, respectively, 60% and 40% of Pt found. This study demonstrates that in the first hours after its infusion, oxaliplatin, in addition to other Pt compounds, is present in plasma and urine and that Pt is bound to albumin, gamma globulins, and hemoglobin. PMID- 11038168 TI - The specificity of glucuronidation of entacapone and tolcapone by recombinant human UDP-glucuronosyltransferases. AB - The COMT inhibitors entacapone and tolcapone are rapidly metabolized in vivo, mainly by glucuronidation. In this work, the main UGT isoforms responsible for their glucuronidation in vitro were characterized by using a subset of representative cloned and expressed human UGT isoforms. Entacapone in particular was seen to be an exceptionally good substrate for UGT1A9 with an even higher reaction velocity value at 500 microM substrate concentration compared with that of the commonly used substrate, propofol (1.3 and 0.78 nmol min(-1) mg(-1), respectively). Neither entacapone nor tolcapone was glucuronidated by UGT1A6. Tolcapone was not detectably glucuronidated by UGT1A1, and the rate of glucuronidation of entacapone was also low by this isoform. However, UGT1A1 was the only UGT capable of catalyzing the formation of two glucuronides of the catecholic entacapone. Both COMT inhibitors were glucuronidated at low rates by the representative members of the UGT2B family, UGT2B7 and UGT2B15. Michaelis Menten parameters were determined for entacapone and tolcapone using recombinant human UGT isoforms and human liver microsomes to compare the kinetic properties of the two COMT inhibitors. The kinetic data illustrates that UGT1A9 exhibited a much greater rate of glucuronidation and a far lower K(m) value for both entacapone and tolcapone than UGT2B15 and UGT2B7 whose contribution is minor by comparison. Entacapone showed a 3 to 4 times higher V(max) value and a 4 to 6 times lower K(m) value compared with those of tolcapone both in UGT1A9 cell lysates and in human liver microsomes. PMID- 11038169 TI - Selective accumulation of raft-associated membrane protein LAT in T cell receptor signaling assemblies. AB - Activation of T cell antigen receptor (TCR) induces tyrosine phosphorylations that mediate the assembly of signaling protein complexes. Moreover, cholesterol sphingolipid raft membrane domains have been implicated to play a role in TCR signal transduction. Here, we studied the assembly of TCR with signal transduction proteins and raft markers in plasma membrane subdomains of Jurkat T leukemic cells. We employed a novel method to immunoisolate plasma membrane subfragments that were highly concentrated in activated TCR-CD3 complexes and associated signaling proteins. We found that the raft transmembrane protein linker for activation of T cells (LAT), but not a palmitoylation-deficient non raft LAT mutant, strongly accumulated in TCR-enriched immunoisolates in a tyrosine phosphorylation-dependent manner. In contrast, other raft-associated molecules, including protein tyrosine kinases Lck and Fyn, GM1, and cholesterol, were not highly concentrated in TCR-enriched plasma membrane immunoisolates. Many downstream signaling proteins coisolated with the TCR/LAT-enriched plasma membrane fragments, suggesting that LAT/TCR assemblies form a structural scaffold for TCR signal transduction proteins. Our results indicate that TCR signaling assemblies in plasma membrane subdomains, rather than generally concentrating raft-associated membrane proteins and lipids, form by a selective protein mediated anchoring of the raft membrane protein LAT in vicinity of TCR. PMID- 11038170 TI - Ectodomain shedding of epidermal growth factor receptor ligands is required for keratinocyte migration in cutaneous wound healing. AB - Keratinocyte proliferation and migration are essential to cutaneous wound healing and are, in part, mediated in an autocrine fashion by epidermal growth factor receptor (EGFR)-ligand interactions. EGFR ligands are initially synthesized as membrane-anchored forms, but can be processed and shed as soluble forms. We provide evidence here that wound stimuli induce keratinocyte shedding of EGFR ligands in vitro, particularly the ligand heparin-binding EGF-like growth factor (HB-EGF). The resulting soluble ligands stimulated transient activation of EGFR. OSU8-1, an inhibitor of EGFR ligand shedding, abrogated the wound-induced activation of EGFR and caused suppression of keratinocyte migration in vitro. Soluble EGFR-immunoglobulin G-Fcgamma fusion protein, which is able to neutralize all EGFR ligands, also suppressed keratinocyte migration in vitro. The application of OSU8-1 to wound sites in mice greatly retarded reepithelialization as the result of a failure in keratinocyte migration, but this effect could be overcome if recombinant soluble HB-EGF was added along with OSU8-1. These findings indicate that the shedding of EGFR ligands represents a critical event in keratinocyte migration, and suggest their possible use as an effective clinical treatment in the early phases of wound healing. PMID- 11038171 TI - A novel member of the netrin family, beta-netrin, shares homology with the beta chain of laminin: identification, expression, and functional characterization. AB - The netrins are a family of laminin-related molecules. Here, we characterize a new member of the family, beta-netrin. beta-Netrin is homologous to the NH(2) terminus of laminin chain short arms; it contains a laminin-like domain VI and 3.5 laminin EGF repeats and a netrin C domain. Unlike other netrins, this new netrin is more related to the laminin beta chains, thus, its name beta-netrin. An initial analysis of the tissue distribution revealed that kidney, heart, ovary, retina, and the olfactory bulb were tissues of high expression. We have expressed the molecule in a eukaryotic cell expression system and made antibodies to the expressed product. Both in situ hybridization and immunohistochemistry were used to describe the cellular source of beta-netrin and where beta-netrin is deposited. beta-Netrin is a basement membrane component; it is present in the basement membranes of the vasculature, kidney, and ovaries. In addition, beta netrin is expressed in a limited set of fiber tracts within the brain, including the lateral olfactory tract and the vomeronasal nerve. Functional studies were performed and show that beta-netrin promotes neurite elongation from olfactory bulb explants. Together, these data suggest that beta-netrin is important in neural, kidney, and vascular development. PMID- 11038172 TI - Indications for a novel muscular dystrophy pathway. gamma-filamin, the muscle specific filamin isoform, interacts with myotilin. AB - gamma-Filamin, also called ABP-L, is a filamin isoform that is specifically expressed in striated muscles, where it is predominantly localized in myofibrillar Z-discs. A minor fraction of the protein shows subsarcolemmal localization. Although gamma-filamin has the same overall structure as the two other known isoforms, it is the only isoform that carries a unique insertion in its immunoglobulin (Ig)-like domain 20. Sequencing of the genomic region encoding this part of the molecule shows that this insert is encoded by an extra exon. Transient transfections of the insert-bearing domain in skeletal muscle cells and cardiomyocytes show that this single domain is sufficient for targeting to developing and mature Z-discs. The yeast two-hybrid method was used to identify possible binding partners for the insert-bearing Ig-like domain 20 of gamma filamin. The two Ig-like domains of the recently described alpha-actinin-binding Z-disc protein myotilin were found to interact directly with this filamin domain, indicating that the amino-terminal end of gamma-filamin may be indirectly anchored to alpha-actinin in the Z-disc via myotilin. Since defects in the myotilin gene were recently reported to cause a form of autosomal dominant limb girdle muscular dystrophy, our findings provide a further contribution to the molecular understanding of this disease. PMID- 11038173 TI - Role of the PI3K regulatory subunit in the control of actin organization and cell migration. AB - Cell migration represents an important cellular response that utilizes cytoskeletal reorganization as its driving force. Here, we describe a new signaling cascade linking PDGF receptor stimulation to actin rearrangements and cell migration. We demonstrate that PDGF activates Cdc42 and its downstream effector N-WASP to mediate filopodia formation, actin stress fiber disassembly, and a reduction in focal adhesion complexes. Induction of the Cdc42 pathway is independent of phosphoinositide 3-kinase (PI3K) enzymatic activity, but it is dependent on the p85alpha regulatory subunit of PI3K. Finally, data are provided showing that activation of this pathway is required for PDGF-induced cell migration on collagen. These observations show the essential role of the PI3K regulatory subunit p85alpha in controlling PDGF receptor-induced cytoskeletal changes and cell migration, illustrating a novel signaling pathway that links receptor stimulation at the cell membrane with actin dynamics. PMID- 11038174 TI - The reversible modification regulates the membrane-binding state of Apg8/Aut7 essential for autophagy and the cytoplasm to vacuole targeting pathway. AB - Autophagy and the Cvt pathway are examples of nonclassical vesicular transport from the cytoplasm to the vacuole via double-membrane vesicles. Apg8/Aut7, which plays an important role in the formation of such vesicles, tends to bind to membranes in spite of its hydrophilic nature. We show here that the nature of the association of Apg8 with membranes changes depending on a series of modifications of the protein itself. First, the carboxy-terminal Arg residue of newly synthesized Apg8 is removed by Apg4/Aut2, a novel cysteine protease, and a Gly residue becomes the carboxy-terminal residue of the protein that is now designated Apg8FG. Subsequently, Apg8FG forms a conjugate with an unidentified molecule "X" and thereby binds tightly to membranes. This modification requires the carboxy-terminal Gly residue of Apg8FG and Apg7, a ubiquitin E1-like enzyme. Finally, the adduct Apg8FG-X is reversed to soluble or loosely membrane-bound Apg8FG by cleavage by Apg4. The mode of action of Apg4, which cleaves both newly synthesized Apg8 and modified Apg8FG, resembles that of deubiquitinating enzymes. A reaction similar to ubiquitination is probably involved in the second modification. The reversible modification of Apg8 appears to be coupled to the membrane dynamics of autophagy and the Cvt pathway. PMID- 11038175 TI - Characterization of the signal that directs Tom20 to the mitochondrial outer membrane. AB - Tom20 is a major receptor of the mitochondrial preprotein translocation system and is bound to the outer membrane through the NH(2)-terminal transmembrane domain (TMD) in an Nin-Ccyt orientation. We analyzed the mitochondria-targeting signal of rat Tom20 (rTom20) in COS-7 cells, using green fluorescent protein (GFP) as the reporter by systematically introducing deletions or mutations into the TMD or the flanking regions. Moderate TMD hydrophobicity and a net positive charge within five residues of the COOH-terminal flanking region were both critical for mitochondria targeting. Constructs without net positive charges within the flanking region, as well as those with high TMD hydrophobicity, were targeted to the ER-Golgi compartments. Intracellular localization of rTom20-GFP fusions, determined by fluorescence microscopy, was further verified by cell fractionation. The signal recognition particle (SRP)-induced translation arrest and photo-cross-linking demonstrated that SRP recognized the TMD of rTom20-GFP, but with reduced affinity, while the positive charge at the COOH-terminal flanking segment inhibited the translation arrest. The mitochondria-targeting signal identified in vivo also functioned in the in vitro system. We conclude that NH(2)-terminal TMD with a moderate hydrophobicity and a net positive charge in the COOH-terminal flanking region function as the mitochondria-targeting signal of the outer membrane proteins, evading SRP-dependent ER targeting. PMID- 11038176 TI - TRAPP stimulates guanine nucleotide exchange on Ypt1p. AB - TRAPP, a novel complex that resides on early Golgi, mediates the targeting of ER to-Golgi vesicles to the Golgi apparatus. Previous studies have shown that YPT1, which encodes the small GTP-binding protein that regulates membrane traffic at this stage of the secretory pathway, interacts genetically with BET3 and BET5. Bet3p and Bet5p are 2 of the 10 identified subunits of TRAPP. Here we show that TRAPP preferentially binds to the nucleotide-free form of Ypt1p. Mutants with defects in several TRAPP subunits are temperature-sensitive in their ability to displace GDP from Ypt1p. Furthermore, the purified TRAPP complex accelerates nucleotide exchange on Ypt1p. Our findings imply that Ypt1p, which is present on ER-to-Golgi transport vesicles, is activated at the Golgi once it interacts with TRAPP. PMID- 11038177 TI - Sorting of yeast membrane proteins into an endosome-to-Golgi pathway involves direct interaction of their cytosolic domains with Vps35p. AB - Resident late-Golgi membrane proteins in Saccharomyces cerevisiae are selectively retrieved from a prevacuolar-endosomal compartment, a process dependent on aromatic amino acid-based sorting determinants on their cytosolic domains. The formation of retrograde vesicles from the prevacuolar compartment and the selective recruitment of vesicular cargo are thought to be mediated by a peripheral membrane retromer protein complex. We previously described mutations in one of the retromer subunit proteins, Vps35p, which caused cargo-specific defects in retrieval. By genetic and biochemical means we now show that Vps35p directly associates with the cytosolic domains of cargo proteins. Chemical cross linking, followed by coimmunoprecipitation, demonstrated that Vps35p interacts with the cytosolic domain of A-ALP, a model late-Golgi membrane protein, in a retrieval signal-dependent manner. Furthermore, mutations in the cytosolic domains of A-ALP and another cargo protein, Vps10p, were identified that suppressed cargo-specific mutations in Vps35p but did not suppress the retrieval defects of a vps35 null mutation. Suppression was shown to be due to an improvement in protein sorting at the prevacuolar compartment. These data strongly support a model in which Vps35p acts as a "receptor" protein for recognition of the retrieval signal domains of cargo proteins during their recruitment into retrograde vesicles. PMID- 11038178 TI - Decreased c-Src expression enhances osteoblast differentiation and bone formation. AB - c-src deletion in mice leads to osteopetrosis as a result of reduced bone resorption due to an alteration of the osteoclast. We report that deletion/reduction of Src expression enhances osteoblast differentiation and bone formation, contributing to the increase in bone mass. Bone histomorphometry showed that bone formation was increased in Src null compared with wild-type mice. In vitro, alkaline phosphatase (ALP) activity and nodule mineralization were increased in primary calvarial cells and in SV40-immortalized osteoblasts from Src(-/-) relative to Src(+/+) mice. Src-antisense oligodeoxynucleotides (AS src) reduced Src levels by approximately 60% and caused a similar increase in ALP activity and nodule mineralization in primary osteoblasts in vitro. Reduction in cell proliferation was observed in primary and immortalized Src(-/-) osteoblasts and in normal osteoblasts incubated with the AS-src. Semiquantitative reverse transcriptase-PCR revealed upregulation of ALP, Osf2/Cbfa1 transcription factor, PTH/PTHrP receptor, osteocalcin, and pro-alpha 2(I) collagen in Src-deficient osteoblasts. The expression of the bone matrix protein osteopontin remained unchanged. Based on these results, we conclude that the reduction of Src expression not only inhibits bone resorption, but also stimulates osteoblast differentiation and bone formation, suggesting that the osteogenic cells may contribute to the development of the osteopetrotic phenotype in Src-deficient mice. PMID- 11038179 TI - Selective disruption of nuclear import by a functional mutant nuclear transport carrier. AB - p10/NTF2 is a nuclear transport carrier that mediates the uptake of cytoplasmic RanGDP into the nucleus. We constructed a point mutant of p10, D23A, that exhibited unexpected behavior both in digitonin-permeabilized and microinjected mammalian cells. D23A p10 was markedly more efficient than wild-type (wt) p10 at supporting Ran import, but simultaneously acted as a dominant-negative inhibitor of classical nuclear localization sequence (cNLS)-mediated nuclear import supported by karyopherins (Kaps) alpha and beta1. Binding studies indicated that these two nuclear transport carriers of different classes, p10 and Kap-beta1, compete for identical and/or overlapping binding sites at the nuclear pore complex (NPC) and that D23A p10 has an increased affinity relative to wt p10 and Kap-beta1 for these shared binding sites. Because of this increased affinity, D23A p10 is able to import its own cargo (RanGDP) more efficiently than wt p10, but Kap-beta1 can no longer compete efficiently for shared NPC docking sites, thus the import of cNLS cargo is inhibited. The competition of different nuclear carriers for shared NPC docking sites observed here predicts a dynamic equilibrium between multiple nuclear transport pathways inside the cell that could be easily shifted by a transient modification of one of the carriers. PMID- 11038180 TI - Gag3p, an outer membrane protein required for fission of mitochondrial tubules. AB - Mitochondrial morphology and function depend on MGM1, a Saccharomyces cerevisiae gene encoding a dynamin-like protein of the mitochondrial outer membrane. Here, we show that mitochondrial fragmentation and mitochondrial genome loss caused by lesions in MGM1 are suppressed by three novel mutations, gag1, gag2, and gag3 (for glycerol-adapted growth). Cells with any of the gag mutations displayed aberrant mitochondrial morphology characterized by elongated, unbranched tubes and highly fenestrated structures. Additionally, each of the gag mutations prevented mitochondrial fragmentation caused by loss of the mitochondrial fusion factor, Fzo1p, or by treatment of cells with sodium azide. The gag1 mutation mapped to DNM1 that encodes a dynamin-related protein required for mitochondrial fission. GAG3 encodes a novel WD40-repeat protein previously found to interact with Dnm1p in a two-hybrid assay. Gag3p was localized to mitochondria where it was found to associate as a peripheral protein on the cytosolic face of the outer membrane. This association requires neither the DNM1 nor GAG2 gene products. However, the localization of Dnm1p to the mitochondrial outer membrane is substantially reduced by the gag2 mutation, but unaffected by loss of Gag3p. These results indicate that Gag3p plays a distinct role on the mitochondrial surface to mediate the fission of mitochondrial tubules. PMID- 11038181 TI - The dynamin-related GTPase, Mgm1p, is an intermembrane space protein required for maintenance of fusion competent mitochondria. AB - Mutations in the dynamin-related GTPase, Mgm1p, have been shown to cause mitochondrial aggregation and mitochondrial DNA loss in Saccharomyces cerevisiae cells, but Mgm1p's exact role in mitochondrial maintenance is unclear. To study the primary function of MGM1, we characterized new temperature sensitive MGM1 alleles. Examination of mitochondrial morphology in mgm1 cells indicates that fragmentation of mitochondrial reticuli is the primary phenotype associated with loss of MGM1 function, with secondary aggregation of mitochondrial fragments. This mgm1 phenotype is identical to that observed in cells with a conditional mutation in FZO1, which encodes a transmembrane GTPase required for mitochondrial fusion, raising the possibility that Mgm1p is also required for fusion. Consistent with this idea, mitochondrial fusion is blocked in mgm1 cells during mating, and deletion of DNM1, which encodes a dynamin-related GTPase required for mitochondrial fission, blocks mitochondrial fragmentation in mgm1 cells. However, in contrast to fzo1 cells, deletion of DNM1 in mgm1 cells restores mitochondrial fusion during mating. This last observation indicates that despite the phenotypic similarities observed between mgm1 and fzo1 cells, MGM1 does not play a direct role in mitochondrial fusion. Although Mgm1p was recently reported to localize to the mitochondrial outer membrane, our studies indicate that Mgm1p is localized to the mitochondrial intermembrane space. Based on our localization data and Mgm1p's structural homology to dynamin, we postulate that it functions in inner membrane remodeling events. In this context, the observed mgm1 phenotypes suggest that inner and outer membrane fission is coupled and that loss of MGM1 function may stimulate Dnm1p-dependent outer membrane fission, resulting in the formation of mitochondrial fragments that are structurally incompetent for fusion. PMID- 11038182 TI - Mdv1p is a WD repeat protein that interacts with the dynamin-related GTPase, Dnm1p, to trigger mitochondrial division. AB - Mitochondrial fission is mediated by the dynamin-related GTPase, Dnm1p, which assembles on the mitochondrial outer membrane into punctate structures associated with sites of membrane constriction and fission. We have identified additional nuclear genes required for mitochondrial fission, termed MDV (for mitochondrial division). MDV1 encodes a predicted soluble protein, containing a coiled-coil motif and seven COOH-terminal WD repeats. Genetic and two-hybrid analyses indicate that Mdv1p interacts with Dnm1p to mediate mitochondrial fission. In addition, Mdv1p colocalizes with Dnm1p in fission-mediating punctate structures on the mitochondrial outer membrane. Whereas localization of Mdv1p to these structures requires Dnm1p, localization of Mdv1p to mitochondrial membranes does not. This indicates that Mdv1p possesses a Dnm1p-independent mitochondrial targeting signal. Dnm1p-independent targeting of Mdv1p to mitochondria requires MDV2. Our data indicate that MDV2 also functions separately to regulate the assembly of Dnm1p into punctate structures. In contrast, Mdv1p is not required for the assembly of Dnm1p, but Dnm1p-containing punctate structures lacking Mdv1p are not able to complete division. Our studies suggest that mitochondrial fission is a multi-step process in which Mdv2p regulates the assembly of Dnm1p into punctate structures and together with Mdv1p functions later during fission to facilitate Dnm1p-dependent mitochondrial membrane constriction and/or division. PMID- 11038183 TI - Dnm1p GTPase-mediated mitochondrial fission is a multi-step process requiring the novel integral membrane component Fis1p. AB - Yeast Dnm1p is a soluble, dynamin-related GTPase that assembles on the outer mitochondrial membrane at sites where organelle division occurs. Although these Dnm1p-containing complexes are thought to trigger constriction and fission, little is known about their composition and assembly, and molecules required for their membrane recruitment have not been isolated. Using a genetic approach, we identified two new genes in the fission pathway, FIS1 and FIS2. FIS1 encodes a novel, outer mitochondrial membrane protein with its amino terminus exposed to the cytoplasm. Fis1p is the first integral membrane protein shown to participate in a eukaryotic membrane fission event. In a related study (Tieu, Q., and J. Nunnari. 2000. J. Cell Biol. 151:353-365), it was shown that the FIS2 gene product (called Mdv1p) colocalizes with Dnm1p on mitochondria. Genetic and morphological evidence indicate that Fis1p, but not Mdv1p, function is required for the proper assembly and distribution of Dnm1p-containing fission complexes on mitochondrial tubules. We propose that mitochondrial fission in yeast is a multi step process, and that membrane-bound Fis1p is required for the proper assembly, membrane distribution, and function of Dnm1p-containing complexes during fission. PMID- 11038184 TI - Targeted ablation of the murine involucrin gene. AB - Involucrin is synthesized in abundance during terminal differentiation of keratinocytes. Involucrin is a substrate for transglutaminase and one of the precursors of the cross-linked envelopes present in the corneocytes of the epidermis and other stratified squamous epithelia. These envelopes make an important contribution to the physical resistance of the epidermis. We have generated mice lacking involucrin from embryonic stem cells whose involucrin gene had been ablated by homologous recombination. These mice developed normally, possessed apparently normal epidermis and hair follicles, and made cornified envelopes that could not be distinguished from those of wild-type mice. No compensatory increase of mRNA for other envelope precursors was observed. PMID- 11038185 TI - Lessons from loricrin-deficient mice: compensatory mechanisms maintaining skin barrier function in the absence of a major cornified envelope protein. AB - The epidermal cornified cell envelope (CE) is a complex protein-lipid composite that replaces the plasma membrane of terminally differentiated keratinocytes. This lamellar structure is essential for the barrier function of the skin and has the ability to prevent the loss of water and ions and to protect from environmental hazards. The major protein of the epidermal CE is loricrin, contributing approximately 70% by mass. We have generated mice that are deficient for this protein. These mice showed a delay in the formation of the skin barrier in embryonic development. At birth, homozygous mutant mice weighed less than control littermates and showed skin abnormalities, such as congenital erythroderma with a shiny, translucent skin. Tape stripping experiments suggested that the stratum corneum stability was reduced in newborn Lor(-/-) mice compared with wild-type controls. Isolated mutant CEs were more easily fragmented by sonication in vitro, indicating a greater susceptibility to mechanical stress. Nevertheless, we did not detect impaired epidermal barrier function in these mice. Surprisingly, the skin phenotype disappeared 4-5 d after birth. At least one of the compensatory mechanisms preventing a more severe skin phenotype in newborn Lor(-/-) mice is an increase in the expression of other CE components, such as SPRRP2D and SPRRP2H, members of the family of "small proline rich proteins", and repetin, a member of the "fused gene" subgroup of the S100 gene family. PMID- 11038186 TI - Transgenic mice expressing a mutant form of loricrin reveal the molecular basis of the skin diseases, Vohwinkel syndrome and progressive symmetric erythrokeratoderma. AB - Mutations in the cornified cell envelope protein loricrin have been reported recently in some patients with Vohwinkel syndrome (VS) and progressive symmetric erythrokeratoderma (PSEK). To establish a causative relationship between loricrin mutations and these diseases, we have generated transgenic mice expressing a COOH terminal truncated form of loricrin that is similar to the protein expressed in VS and PSEK patients. At birth, transgenic mice (ML.VS) exhibited erythrokeratoderma with an epidermal barrier dysfunction. 4 d after birth, high expressing transgenic animals showed a generalized scaling of the skin, as well as a constricting band encircling the tail and, by day 7, a thickening of the footpads. Histologically, ML. VS transgenic mice also showed retention of nuclei in the stratum corneum, a characteristic feature of VS and PSEK. Immunofluorescence and immunoelectron microscopy showed the mutant loricrin protein in the nucleus and cytoplasm of epidermal keratinocytes, but did not detect the protein in the cornified cell envelope. Transfection experiments indicated that the COOH-terminal domain of the mutant loricrin contains a nuclear localization signal. To determine whether the ML.VS phenotype resulted from dominant-negative interference of the transgene with endogenous loricrin, we mated the ML.VS transgenics with loricrin knockout mice. A severe phenotype was observed in mice that lacked expression of wild-type loricrin. Since loricrin knockout mice are largely asymptomatic (Koch, P.K., P. A. de Viragh, E. Scharer, D. Bundman, M.A. Longley, J. Bickenbach, Y. Kawachi, Y. Suga, Z. Zhou, M. Huber, et al., J. Cell Biol. 151:389-400, this issue), this phenotype may be attributed to expression of the mutant form of loricrin. Thus, deposition of the mutant protein in the nucleus appears to interfere with late stages of epidermal differentiation, resulting in a VS-like phenotype. PMID- 11038187 TI - Evidence that the transition of HIV-1 gp41 into a six-helix bundle, not the bundle configuration, induces membrane fusion. AB - Many viral fusion proteins exhibit a six-helix bundle as a core structure. HIV Env-induced fusion was studied to resolve whether membrane merger was due to the transition into the bundle configuration or occurred after bundle formation. Suboptimal temperature was used to arrest fusion at an intermediate stage. When bundle formation was prevented by adding inhibitory peptides at this stage, membranes did not merge upon raising temperature. Inversely, when membrane merger was prevented by incorporating lysophosphatidylcholine (LPC) into cell membranes at the intermediate, the bundle did not form upon optimizing temperature. In the absence of LPC, the six-helix bundle did not form when the temperature of the intermediate was raised for times too short to promote fusion. Kinetic measures showed that after the temperature pulse, cells had not advanced further toward fusion. The latter results indicate that bundle formation is the rate-limiting step between the arrested intermediate and fusion. Electrical measures showed that the HIV Env-induced pore is initially large and grows rapidly. It is proposed that bundle formation and fusion are each contingent on the other and that movement of Env during its transition into the six-helix bundle directly induces the lipid rearrangements of membrane fusion. Because peptide inhibition showed that, at the intermediate stage, the heptad repeats of gp41 have become stably exposed, creation of the intermediate could be of importance in drug and/or vaccine development. PMID- 11038188 TI - The transmembrane domain of influenza hemagglutinin exhibits a stringent length requirement to support the hemifusion to fusion transition. AB - Glycosylphosphatidylinositol-anchored influenza hemagglutinin (GPI-HA) mediates hemifusion, whereas chimeras with foreign transmembrane (TM) domains mediate full fusion. A possible explanation for these observations is that the TM domain must be a critical length in order for HA to promote full fusion. To test this hypothesis, we analyzed biochemical properties and fusion phenotypes of HA with alterations in its 27-amino acid TM domain. Our mutants included sequential 2 amino acid (Delta2-Delta14) and an 11-amino acid deletion from the COOH-terminal end, deletions of 6 or 8 amino acids from the NH(2)-terminal and middle regions, and a deletion of 12 amino acids from the NH(2)-terminal end of the TM domain. We also made several point mutations in the TM domain. All of the mutants except Delta14 were expressed at the cell surface and displayed biochemical properties virtually identical to wild-type HA. All the mutants that were expressed at the cell surface promoted full fusion, with the notable exception of deletions of >10 amino acids. A mutant in which 11 amino acids were deleted was severely impaired in promoting full fusion. Mutants in which 12 amino acids were deleted (from either end) mediated only hemifusion. Hence, a TM domain of 17 amino acids is needed to efficiently promote full fusion. Addition of either the hydrophilic HA cytoplasmic tail sequence or a single arginine to Delta12 HA, the hemifusion mutant that terminates with 15 (hydrophobic) amino acids of the HA TM domain, restored full fusion activity. Our data support a model in which the TM domain must span the bilayer to promote full fusion. PMID- 11038189 TI - Ordering the final events in yeast exocytosis. AB - In yeast, assembly of exocytic soluble N-ethylmaleimide-sensitive fusion protein (NSF) attachment protein receptor (SNARE) complexes between the secretory vesicle SNARE Sncp and the plasma membrane SNAREs Ssop and Sec9p occurs at a late stage of the exocytic reaction. Mutations that block either secretory vesicle delivery or tethering prevent SNARE complex assembly and the localization of Sec1p, a SNARE complex binding protein, to sites of secretion. By contrast, wild-type levels of SNARE complexes persist in the sec1-1 mutant after a secretory block is imposed, suggesting a role for Sec1p after SNARE complex assembly. In the sec18-1 mutant, cis-SNARE complexes containing surface-accessible Sncp accumulate in the plasma membrane. Thus, one function of Sec18p is to disassemble SNARE complexes on the postfusion membrane. PMID- 11038190 TI - Geranylgeranylated SNAREs are dominant inhibitors of membrane fusion. AB - Exocytosis in yeast requires the assembly of the secretory vesicle soluble N ethylmaleimide-sensitive factor attachment protein receptor (v-SNARE) Sncp and the plasma membrane t-SNAREs Ssop and Sec9p into a SNARE complex. High-level expression of mutant Snc1 or Sso2 proteins that have a COOH-terminal geranylgeranylation signal instead of a transmembrane domain inhibits exocytosis at a stage after vesicle docking. The mutant SNARE proteins are membrane associated, correctly targeted, assemble into SNARE complexes, and do not interfere with the incorporation of wild-type SNARE proteins into complexes. Mutant SNARE complexes recruit GFP-Sec1p to sites of exocytosis and can be disassembled by the Sec18p ATPase. Heterotrimeric SNARE complexes assembled from both wild-type and mutant SNAREs are present in heterogeneous higher-order complexes containing Sec1p that sediment at greater than 20S. Based on a structural analogy between geranylgeranylated SNAREs and the GPI-HA mutant influenza virus fusion protein, we propose that the mutant SNAREs are fusion proteins unable to catalyze fusion of the distal leaflets of the secretory vesicle and plasma membrane. In support of this model, the inverted cone-shaped lipid lysophosphatidylcholine rescues secretion from SNARE mutant cells. PMID- 11038191 TI - The triad targeting signal of the skeletal muscle calcium channel is localized in the COOH terminus of the alpha(1S) subunit. AB - The specific localization of L-type Ca(2+) channels in skeletal muscle triads is critical for their normal function in excitation-contraction (EC) coupling. Reconstitution of dysgenic myotubes with the skeletal muscle Ca(2+) channel alpha(1S) subunit restores Ca(2+) currents, EC coupling, and the normal localization of alpha(1S) in the triads. In contrast, expression of the neuronal alpha(1A) subunit gives rise to robust Ca(2+) currents but not to triad localization. To identify regions in the primary structure of alpha(1S) involved in the targeting of the Ca(2+) channel into the triads, chimeras of alpha(1S) and alpha(1A) were constructed, expressed in dysgenic myotubes, and their subcellular distribution was analyzed with double immunofluorescence labeling of the alpha(1S)/alpha(1A) chimeras and the ryanodine receptor. Whereas chimeras containing the COOH terminus of alpha(1A) were not incorporated into triads, chimeras containing the COOH terminus of alpha(1S) were correctly targeted. Mapping of the COOH terminus revealed a triad-targeting signal contained in the 55 amino-acid sequence (1607-1661) proximal to the putative clipping site of alpha(1S). Transferring this triad targeting signal to alpha(1A) was sufficient for targeting and clustering the neuronal isoform into skeletal muscle triads and caused a marked restoration of Ca(2+)-dependent EC coupling. PMID- 11038192 TI - A mitochondrial division apparatus takes shape. PMID- 11038193 TI - The complexity and redundancy of epithelial barrier function. PMID- 11038195 TI - Nuclear medicine applications for neuroendocrine tumors. AB - Sensitive, specific radiopharmaceuticals are available for scintigraphic diagnosis and internal radiotherapy of neuroendocrine tumors. (123)I-MIBG (metaiodobenzylguanidine) scintigraphy is the examination of choice for visualizing tumor sites of pheochromocytoma. In the event of malignant pheochromocytoma or carcinoid tumor, this examination allows assessment of the presence or absence of tumor uptake and can guide radiotherapy with (131)I-MIBG. The peptides secreted by neuroendocrine tumors can be radiolabeled for targeting of their specific receptors. Scintigraphy using a (111)In-labeled somatostatin analog (octreotide) is the examination of choice for diagnosis of the spread of gastroenteropancreatic and carcinoid tumors, as it is more sensitive than morphologic imaging techniques. It can also guide radiotherapy performed with the same pharmaceutical vector. These same two agents (MIBG and octreotide) can be used therapeutically by replacing (123)I with (131)I and (111)In by (90)Y. A transient palliative effect is obtained for a variable number of tumors (most often large ones) that take up the radiopharmaceutic agent well. There is general consensus that, for relatively radioresistant solid tumors, this type of radiotherapy is efficient only in the event of small tumor targets (a few millimeters in diameter) whose uptake is maximal, allowing more homogeneous distribution than that achieved with large tumors. Thus for optimal control of the disease it is recommended first to use scintigraphic imaging to confirm that the tumor takes up the radiopharmaceutical agent in question ((123)I-MIBG or (111)In-octreotide) and then reduce the tumor burden surgically before injecting high therapeutic activity (possibly with reinjection of peripheral stem cells). This treatment can be repeated three times every 3 months before evaluating the response. In these conditions, internal radiotherapy can be beneficial or even determinant for controlling disease progression. PMID- 11038194 TI - HIV-1 membrane fusion: targets of opportunity. PMID- 11038196 TI - Impact of initial surgical treatment on survival of patients with differentiated thyroid cancer: experience of an endocrine surgery center in an iodine-deficient region. AB - This retrospective clinical study was designed to analyze the impact of the initial surgical procedure on the survival of 1000 patients with differentiated thyroid cancer of follicular cell origin who had a thyroid operation and were followed for the 30 years between 1968 and 1998 (median 14 years) in an iodine deficient region where goiter is endemic. There were 753 women and 247 men with a mean age of 42.8 +/- 6.7 years (range 17-86 years). Patients were divided into three groups. All patients had undergone thyroxine treatment and thyroid stimulating hormone (TSH) suppression, and most had had iodine-131 treatment postoperatively. Group A consisted of 336 patients with differentiated thyroid cancer (DTC) who were treated with bilateral subtotal thyroidectomy in our institution or elsewhere. Group B consisted of 158 patients with DTC who were treated initially with unilateral total lobectomy and contralateral subtotal lobectomy in our institution or elsewhere and underwent reoperation in our department. Group C consisted of 506 patients with DTC who were treated initially with total or near-total thyroidectomy in our department. Kaplan-Meyer survival analysis was used. Recurrence was seen in 23% and death in 8% of the patients. The 20-year survival rates were 76%, 85%, and 92% for groups A, B, and C, respectively. The survival difference among the patients of group A and groups B and C was found to be statistically different (p < 0.001). Long-term survival of patients with differentiated thyroid cancer living in endemic areas for goiter can be influenced by the initial surgical treatment. Patients treated initially with total or near-total thyroidectomy appear to have a better prognosis. PMID- 11038197 TI - Total thyroidectomy or thyroid lobectomy in patients with low-risk differentiated thyroid cancer: surgical decision analysis of a controversy using a mathematical model. AB - There is a general consensus that total or near-total thyroidectomy is the optimal treatment for patients with high risk differentiated thyroid cancer (DTC), but the optimal extent of thyroidectomy in patients with low risk DTC continues to be controversial. To determine the optimal extent of thyroidectomy in patients with low risk DTC, we used decision analysis to compare the trade offs of total thyroidectomy (TT) to thyroid lobectomy (TL). The decision analysis model included the probabilities of thyroidectomy complications, risk of DTC recurrence, and death from DTC. This information was obtained from the literature and from outcome data for patients with low risk DTC from our institution. In addition, the concept of utilities was used in the analysis. To determine the utility of each health outcome state (thyroidectomy complication, DTC recurrence, and DTC mortality for low risk patients) a survey was conducted. Overall, prospective patients viewed DTC recurrence as less desirable than thyroidectomy complication. The utilities assigned by the survey participants varied over a wide range, with 61.5% of the individuals viewing the occurrence of a thyroidectomy complication as better than DTC recurrence. At baseline utilities and probabilities, TT had a higher expected utility than TL. One-way sensitivity analysis varying the rates of (1) thyroidectomy complication, (2) DTC recurrence, and (3) DTC mortality over the possible range showed that complication from initial thyroidectomy was the most important factor that determined the preferred extent of thyroidectomy. TL was the preferred surgical approach only if a complication rate of > 33:1, TT/TL complication rate ratio, was assumed. When no differences in DTC recurrence between the two approaches was assumed in the model, TL had a higher expected utility using the baseline utilities of thyroidectomy complication and DTC mortality. The analysis indicates that TT in patients with low risk DTC is preferable to TL. However, TL is preferred if (1) no difference in the DTC recurrence rate between the two approaches is assumed, (2) a higher complication rate for TT is used (> 33 times higher), or (3) the utility ratio of thyroidectomy complication to DTC recurrence is < 0.8 TL. We believe this decision analysis model provides an objective approach that others can use to select the optimal extent of thyroidectomy based on patient preference of health outcome states, institution-specific outcome data for DTC recurrence or mortality, and the surgeon-specific complication rate. PMID- 11038198 TI - Surgery for Graves' disease: total versus subtotal thyroidectomy-results of a prospective randomized trial. AB - The effect of surgery on Graves' orbitopathy (GO) is still controversial. Retrospective analyses of many authors (including our own group) demonstrated GO improvement after subtotal thyroid resection in up to 70% of operated patients, so the question arose whether total thyroidectomy could add anything to this pronounced positive effect on GO. We therefore performed a prospective randomized trial on 150 patients with Graves' disease (125 women, 25 men; mean thyroid volume 80.5 ml) comparing three surgical procedures (bilateral subtotal thyroid resection-total remnant < 4 ml; unilateral hemithyroidectomy with contralateral subtotal thyroid resection-remnant < 4 ml; total thyroidectomy) and their effect on postoperative GO changes, postoperative thyroid-stimulating hormone receptor (TSH-R) antibody titers, and postoperative complication rates. After a period of at least 6 months (6-36 months) GO had improved in 71% to 74% of all patients regardless of whether total or subtotal thyroidectomy was performed. TSH-R antibody titers showed no differences for the three surgical groups. Postoperative recurrent hyperthyroidism occurred in two patients with subtotal resections, and early postoperative hypoparathyroidism was more frequently detected in patients with total thyroidectomy than in those with subtotal thyroid resection (28% vs. 12%; p < 0.002). In respect to possible postoperative hypoparathyroidism and a lack of difference in postoperative GO changes, we do not advocate total thyroidectomy for patients with Graves' disease and Graves' orbitopathy but prefer radical subtotal thyroid resection with a remnant of less than 4 ml. PMID- 11038199 TI - Is the new classification of neuroendocrine pancreatic tumors of clinical help? AB - A new concept of classifying neuroendocrine pancreatic tumors based on clinicopathologic patterns was summarized recently. To evaluate the clinical reliability and prognostic specificity of this classification system, 100 neuroendocrine pancreatic tumors were retrospectively categorized as "benign," "uncertain," and "malignant" based on tumor risk factors (size, local invasion and angioinvasion, cell atypia, metastases) and were followed for disease recurrence and progression. Altogether, 71 functioning tumors (insulinoma, gastrinoma, glucagonoma, enterochromaffin-like (ECL)oma, somatostatinoma) and 29 nonfunctioning neuroendocrine pancreatic tumors (NETs) were studied. NETs had an increased risk of malignancy (p < 0.05). Tumor size, gross invasion, and metastases correlated significantly with tumor behavior and allowed us to distinguish between "benign" and "malignant" tumors. About 89% of the tumors < or = 20 mm were "benign," whereas 71% > 20 mm were "malignant" (p < 0.05). In patients with "benign" and "uncertain" neuroendocrine pancreatic tumors, neither recurrence nor progression of disease was seen. About 41% of the patients with "malignant" tumors died of the disease. The 5-year estimated cumulative survival of those with "benign" and "uncertain" tumors was 100% and 52 +/- 10% for those with "malignant" tumors (p < 0.05). Histomorphologic details classifying the behavior of an "uncertain" tumor are known only after initial treatment and definitive histopathologic investigation. Thus this information is of limited clinical help for treatment strategies. PMID- 11038200 TI - Intraoperative parathyroid aspiration and parathyroid hormone assay as an alternative to frozen section for tissue identification. AB - Most people would agree that successful parathyroidectomy depends on two important variables: the surgeon's recognition and excision of the abnormal parathyroid gland(s) and the pathologist's confirmation that the removed tissue is parathyroid tissue. Frozen section is usually employed to confirm the identity of parathyroid tissue, but occasionally confirmation cannot be made without a permanent section, as with intrathyroidal glands. This study proposes a new method of expeditious and easy confirmation of parathyroid tissue utilizing the immunoassay for quick measurement of intraoperative parathyroid hormone (IOPTH). By directly aspirating the suspected adenoma, the assay becomes a rapid diagnostic tool that can be used as an alternative to frozen section. In cases where the surgeon is already planning to employ the assay, the elimination of frozen section is cost-effective. Intraoperative aspiration of histologically confirmed parathyroid adenomas was performed on 12 consecutive patients undergoing parathyroid surgery. Parathyroid glands were aspirated with a 22-gauge syringe after gland excision. Aspirates were placed in 1 to 3 ml of buffered saline. A similar process was performed on 12 thyroid controls. Specimens were centrifuged, aliquotted, and stored at -70 degrees C. The parathyroid hormone value was analyzed electively by rapid assay and the values recorded. For all parathyroid aspirates, the rapid assay value was > 1500 pg/ml, exceeding the uppermost limit of the diagnostic chart. Values for thyroid aspirates ranged from 58 to 85 pg/ml (mean 75.7 pg/ml). In all cases tissue confirmation was achieved with permanent section. Values were 100% sensitive and specific. Measurement of PTH from intraoperative aspiration of suspected parathyroid adenomas is clinically useful in patients for whom frozen section would routinely be employed. Values > 1500 pg/ml secure the tissue diagnosis. There is no additional cost in cases where IOPTH monitoring is already being utilized to confirm cure. The elimination of frozen section could be cost-effective and, for some institutions, actually decrease the operating time as the IOPTH assay takes only 15 minutes. PTH assay is an accurate diagnostic technique and to date is 100% sensitive and specific for differentiating between parathyroid tumors and thyroid nodules. PMID- 11038201 TI - Postoperative elevated serum levels of intact parathyroid hormone after surgery for parathyroid adenoma: sign of bone remineralization and decreased calcium absorption. AB - Increased levels of intact parathyroid hormone (PTH) have been documented after surgery for primary hyperparathyroidism (pHPT) despite normocalcemia. The pathogenesis remains to be elucidated. Seventeen consecutive patients operated on for solitary parathyroid adenoma were investigated before and at 8 weeks and 1 year after surgery with serum levels of intact PTH, biochemical variables known to reflect PTH activity, and bone mineral content (BMC). In addition, an oral calcium loading test was performed 8 weeks after the operation. All patients had low or normal serum calcium levels during follow-up. Eight weeks after operation six patients (35%) had an increased serum PTH level. These patients (group I) preoperatively had higher serum levels of PTH and alkaline phosphatase than patients with normal PTH levels (group II). They also had lower BMC and larger parathyroid adenomas. They did not differ in renal function. At 8 weeks after operation group I showed higher mean serum levels of osteocalcin and propeptide of type I procollagen but lower urinary calcium excretion. In contrast to patients in group II, they also showed a lower calciuric response and a trend to a lower calcemic response during the oral calcium load. The two groups showed similar parathyroid sensitivity for calcium. Patients in group I demonstrated a significant increase in BMC the first year after the operation. Increased serum PTH 8 weeks after surgery for sporadic parathyroid adenoma was not due to persistent pHPT or impaired renal function. Instead, the results imply there is diminished calcium absorption and increased bone turnover with cortical bone remineralization. PMID- 11038202 TI - Supernumerary parathyroid glands: frequency and surgical significance in treatment of renal hyperparathyroidism. AB - Supernumerary parathyroid glands (SPGs) are found in 13% of random autopsies. The high incidence of SPGs could explain the persistence or trigger recurrence of renal hyperparathyroidism after surgery. The aim of this study was to assess the frequency and clinical relevance of SPG in patients operated on for renal hyperparathyroidism (HPT). In this retrospective study we reviewed the medical records of 290 patients with renal HPT who were initially treated in our department. We examined the anatomic and pathologic findings during cervical surgical exploration and the outcome of HPT during follow-up. SPGs were identified in 87 patients (30%) during the initial cervicotomy, corresponding to intrathymic parathyroid cell islets (one to four) in 70 cases and to extrathymic SPG in 17 patients. Among 260 patients available for follow-up, 11 experienced persistent HPT (4%), and 34 developed recurrent HPT (13%). A total of 25 patients were reoperated on, and SPGs were responsible for 4 of 8 cases of persistent HPT and 4 of 17 cases of recurrent HPT, representing an overall frequency of 32%. The anatomic distribution of SPGs found during reoperations included thymus, retroesophageal grove, carotid sheath, and mediastinum. SPGs are thus present in 30% of patients with renal HPT and are situated mainly in the thymus. Thymectomy should be performed routinely during the first surgical exploration to prevent recurrences arising from anterior mediastinal glands. SPGs were also responsible for 32% of persistent or recurrent HPT. In that setting, frankly ectopic SPGs are not rare, and preoperative imaging appears highly desirable prior to embarking on surgical reexploration. PMID- 11038203 TI - Multivariate analysis of risk factors for postoperative complications in benign goiter surgery: prospective multicenter study in Germany. AB - Risk factors for postoperative complications of benign goiter surgery have not been investigated systematically. To this end, a prospective multicenter study (January 1 through December 31, 1998) was conducted involving 7266 patients with surgery for benign goiter from 45 East German hospitals. High-volume providers (>150 operations per year) performed 69% (5042/7266), intermediate-volume providers 27% (50-150), and low-volume providers 4% (258/7266) of operations. Among the hospital groups, the pattern of thyroid disease did not vary significantly, but there was a trend that small-volume providers tended to perform more operations for uninodular goiter and high-volume providers treated more patients with Graves' disease and recurrent goiter. Extent of resection (p < 0.0001) and remnant size (multinodular goiter and recurrent goiter, p < 0.001), differed significantly, with total thyroidectomy being performed more often in hospitals with more than 150 operations compared to hospitals with an operative volume of less than 150 procedures per year. Despite the larger extent of resection and smaller remnant size, rates of recurrent laryngeal nerve (RLN) palsy or hypoparathyroidism were not increased. When the logistic regression analyses were fitted to evaluate the impact of risk factors on transient and permanent RLN palsy and hypoparathyroidism, larger extent of resection [relative risk (RR) 1.5-2.1] and recurrent goiter (RR 1.8-3.4) consistently evolved as independent risk factors. With hypoparathyroidism, additional significant factors included patient gender (RR 2.1-2.4), hospital operative volume (RR 0.8-1.5), and Graves' disease (RR 2.8). Unlike parathyroid gland identification during hypoparathyroidism, RLN identification (RR 1.6) significantly (p = 0.01) reduced permanent RLN palsy rates. The multivariate analyses clearly confirmed the pivotal role of routine RLN identification, independent of the extent of the thyroid resection. These findings might help hospitals with lower operative volumes to identify patients at increased risk whom they might consider for specialist care. PMID- 11038204 TI - Complications of laparoscopic adrenalectomy: results of 169 consecutive procedures. AB - Laparoscopic adrenalectomy (LA) has become the gold standard for adrenalectomy. Review of the literature indicates that the rate of intra- and postoperative complications is not negligible. The aim of this study was to evaluate the complications observed in a series of 169 consecutive LAs performed at a same center for a variety of endocrine disorders. Between June 1994 and December 1998 a series of 169 LAs were performed in 159 patients: 149 unilateral LAs and 10 bilateral LAs. There were 98 women and 61 men with a mean age of 49. 7 years (range 22-76 years). There were patients with 61 Conn syndrome, 41 with Cushing syndrome, 1 androgen-producing tumor, 29 pheochromocytomas, and 37 nonfunctioning tumors. Mean tumor size was 32 mm (range 7-110 mm). LA was performed by a transperitoneal flank approach in the lateral decubitus position. Mean operating time was 129 minutes (range 48-300 minutes) for unilateral LA and 228 minutes (range 175-275 minutes) for bilateral LA. There was no mortality. Twelve patients had a significant complication (7.5%): three peritoneal hematomas requiring (in two cases) laparotomy and (in one case) transfusion; one parietal hematoma; three intraoperative bleeding episodes without need for transfusion; one partial infarction of the spleen; one pneumothorax; one capsular effraction of the tumor; and two deep venous thromboses. Eight tumors were malignant at final histology (4.7%), of which four were completely removed laparoscopically. Conversion to open surgery was required in eight cases (5%): for malignancy in four cases, difficulty of dissection in three cases, and pneumothorax in one case. With a mean follow-up of 26.58 months (range 6-60 months) all patients are disease-free. We conclude that LA is a safe procedure. With increasing experience the morbidity becomes minor. To avoid complications LA should be converted to open surgery if local invasion is suspected or if there is difficulty with the dissection. PMID- 11038205 TI - Study of three patients with congenital adrenal hyperplasia treated by bilateral adrenalectomy. AB - Medical management of congenital adrenal hyperplasia (CAH) patients has led to suboptimal results in most cases. High glucocorticoid doses, often needed to suppress adrenal androgen production, may lead to signs of Cushing syndrome. Incompletely suppressed androgen levels commonly lead to premature closure of growth centers, acne, virilization, precocious puberty, irregular or absent menses, and decreased fertility in female CAH patients. A newly proposed therapy for CAH patients is bilateral adrenalectomy. Three Caucasian female patients with 21-hydroxylase deficiency were treated with bilateral adrenalectomy. Two of the three procedures were accomplished laparoscopically. In each patient, medical management alone was unsuccessful. Two patients had salt-losing 21-hydroxylase deficiency. The third patient had uncontrolled hyperandrogenism complicated by obesity and glucose intolerance. All patients had low height percentiles with respect to their normalized percentiles for weight. Bone age was advanced in one patient. Androgen and renin levels were well controlled in two patients, whereas the third patient had persistent hyperandrogenism. Bilateral adrenalectomy was performed at the ages of 14, 19, and 30 years with follow-up, to date, of 25 months, 10 months, and 26 months, respectively. Postoperatively, all patients were free from hyperandrogenism. One patient experienced one episode of urosepsis precipitating an addisonian crisis. Bilateral adrenalectomy may successfully address the problems of increasing steroid requirements and hyperandrogenism in patients with severe CAH. The ability to perform this operation laparoscopically coupled with the overall metabolic benefits make bilateral adrenalectomy a reasonable alternative to lifelong androgen suppression in select patients. PMID- 11038206 TI - Surgical strategy for large or malignant endocrine pancreatic tumors. AB - Endocrine pancreatic tumors (EPTs) are rare but have a remarkably better prognosis than adenocarcinoma of the pancreas. Patients with EPTs benefit from surgical and medical therapy, which may alleviate symptoms due to hormonal excess and increase survival. Patients with large or malignant EPTs with infiltrative disease may suffer from local complications, including gastrointestinal bleeding and obstruction and involvement of the superior mesenteric (SMV) and portal (PV) veins. Among 31 patients with operable and large or malignant EPTs, 7 had hormone producing syndromes (insulin, glucagon), and 24 had clinically nonfunctioning EPTs. Surgery in these patients included vascular reconstruction of the SMV/PV (n = 4), resection of infiltrated adjacent organs (n = 5; stomach, transverse colon), or resection of concomitant liver metastases (n = 3). Four patients with conspicuously large insulinomas, and three with glucagonoma were successfully operated on with alleviation of hormonal symptoms. Among the 24 nonfunctioning EPTs, 5 patients had been explored earlier and their tumors judged inoperable due to locally invasive disease or misdiagnosis as pancreatic adenocarcinoma. The operations were performed with no mortality and low morbidity. We conclude that large and malignant EPTs with limited spread of disease may benefit from a combination of medical and surgical therapy. PMID- 11038207 TI - Parathyroid xenotransplantation without immunosuppression in experimental hypoparathyroidism: long-term in vivo function following microencapsulation with a clinically suitable alginate. AB - Permanent hypoparathyroidism is one of the most difficult of all endocrine disorders to treat medically. Because this deficiency syndrome rarely is a life threatening condition, systemic immunosuppression for recipients of transgenic transplants is not justified. An alternative would be protecting the tissue to be transplanted from the immunologic response (immunoisolation) by coating it with a semipermeable membrane- microencapsulation. Unfortunately, prior to the first clinical use, further analysis of the coating substance (alginate) demonstrated that it has potential cancerogenic properties. Using a purified amitogenic alginate suitable for clinical use, we accomplished allotransplantation in a long term animal model and reported the first clinical cases without postoperative immunosuppression recently. In view of the potential clinical use, we investigated the ability of the microencapsulation technology with the novel amitogenic alginate in experimental hypoparathyroidism (80 parathyroidectomized rats) to enable transgenic transplantation across the highest immunologic barrier (xenotransplantation: human to rat) to ensure intact transplant function without immunosuppression. In a controlled, long-term animal study, the effect of microencapsulation on xenotransplanted human parathyroid tissue was evaluated over a period of 30 weeks (microencapsulated and naked hyperplastic parathyroid tissue, respectively). Functionally, human parathyroid tissue was able to replace that of rats. More than 6 months after xenotransplantation 32 of 40 animals that had received microencapsulated transplants were normocalcemic. In contrast, serum calcium concentrations dropped to postparathyroidectomy levels within 3 weeks in the animals that had received naked tissue only. Correspondingly, normocalcemic animals showed vital parathyroid tissue inside the microcapsules, which were surrounded by a small rim of fibroblasts. When combining microencapsulation with an improved tissue culture method, xenotransplantation of human parathyroid tissue and maintenance of its physiologic function is reproducibly achieved over the highest transplantation barrier. Using the amitogenic alginate may be a crucial step toward the first clinical use of this technique for parathyroid xenotransplantation without immunosuppression. PMID- 11038208 TI - Unilateral surgery supported by germline RET oncogene mutation analysis in patients with sporadic medullary thyroid carcinoma. AB - Compared to hereditary medullary thyroid carcinoma (MTC), sporadic MTC tends to be unicentric and confined to one lobe. Patients with sporadic MTC usually undergo total thyroidectomy because of a possible hereditary or bilateral process. We evaluated the usefulness of germline RET oncogene mutation analysis in surgery for apparently sporadic MTC and performed unilateral surgery on patients without detectable mutation. In 36 patients with a preoperative diagnosis of apparently sporadic MTC, we performed germline RET oncogene mutation analyses: before surgery in 8 recent patients and after surgery in 28 who had been treated before 1996. Of the latter, 5 had bilateral MTC. DNA samples were extracted from their peripheral blood, and the polymerase chain reaction products of the RET proto-oncogene were analyzed using single-strand conformation polymorphism analysis and the direct sequencing methods. Before 1996 we often performed total thyroidectomy but changed to hemithyroidectomy thereafter, except in one patient with associated Graves' ophthalmopathy. Our minimal standard practice included systematic central and ipsilateral neck dissection. The outcome was assessed in terms of gastrin- and calcium-stimulated plasma calcitonin levels. Germline RET mutations were found in six patients. Five of these patients had bilateral MTC, whereas all 30 patients without mutation had unilateral disease. Hemithyroidectomy in seven of our recent patients resulted in normalization of plasma calcitonin levels in all, although four were found to have microscopic lymph node involvement. In conclusion, hemithyroidectomy with systematic central and ipsilateral neck dissection is an appropriate procedure for patients with sporadic MTC without detectable germline RET mutations. PMID- 11038209 TI - Occult micro medullary thyroid carcinoma: therapeutic strategy and follow-up. AB - Twenty micro medullary thyroid carcinomas (MTCs) were found in histologic specimens of 19 patients in our department from 1990 to 1998. There were 14 women and 5 men, with a median age of 63 years. The indication for surgery was goiter in 12 patients and a solitary nodule in 7 patients (three differentiated cancers). Altogether, 18 patients had unifocal micro-MTCs with a median diameter of 3.6 mm. One patient had a bilateral MTC (3 and 5 mm, respectively). Surgical procedures consisted of 9 total thyroidectomies and 10 lobectomies or subtotal thyroidectomies. Of these 10 patients, 4 underwent reoperation (totalization). One was operated on 48 months after a positive pentagastrin test: There was no thyroid residual tumor but three lymph node micrometastases. Among the six patients in whom thyroid tissue was left, a 91-year-old woman died of unrelated cause and the five others remain disease-free without biologic abnormalities at follow-ups of 18 to 70 months. Considering the aggressiveness of MTCs, total thyroidectomy with central compartment dissection is theoretically indicated. However, among the nine total thyroidectomies and four secondary totalizations associated with at least central compartment dissection, no other thyroid lesion was observed and only one case of lymph node microinvasion was found. Because of the morbidity associated with reoperation and neck dissection, we propose that it is indicated only for microcarcinomas > 5 mm in diameter, in cases of an abnormal response to pentagastrin, or when it is difficult to ensure prolonged follow-up of the patient. PMID- 11038210 TI - Iatrogenic thyrotoxicosis: causal circumstances, pathophysiology, and principles of treatment-review of the literature. AB - Thyrotoxicosis is the clinical syndrome that results when tissues are exposed to high levels of circulating thyroid hormones. In most instances thyrotoxicosis is due to hyperthyroidism, a term reserved for disorders characterized by overproduction of thyroid hormones by the thyroid gland. Nevertheless, thyrotoxicosis may also result from a variety of conditions other than thyroid hyperfunction. The present report focuses on the etiologies, pathophysiology, and treatment of iatrogenic thyrotoxicosis. Iatrogenic thyrotoxicosis may be caused by (1) subacute thyroiditis (a result of lymphocytic infiltration, cellular injury, trauma, irradiation) with release of preformed hormones into circulation; (2) excessive ingestion of thyroid hormones ("thyrotoxicosis factitia"); (3) iodine-induced hyperthyroidism (radiologic contrast agents, topical antiseptics, other medications). Among these causes of iatrogenic thyrotoxicosis, that induced by the iodine overload and cytotoxicity associated with amiodarone represents a significant challenge. Successful management of amiodarone-induced thyrotoxicosis requires close cooperation between endocrinologists and endocrine surgeons. Surgical treatment may have a leading yet often underestimated role in view of the potential life-threatening severity of this disease, whereas others kinds of iatrogenic thyrotoxicosis are usually treated conservatively. PMID- 11038211 TI - (99m)Tc-sestamibi scintigraphy and cell cycle in parathyroid glands of secondary hyperparathyroidism. AB - Double-phase parathyroid MIBI ((99m)Tc-sestamibi) was performed in 27 patients with secondary hyperparathyroidism (SPT). Focal areas of increased uptake were scored for intensity on a three-point scale. All patients underwent subtotal parathyroidectomy (SPTx), and a total of 78 glands were removed at operation. Blood was obtained from the jugular vein before and after SPTx to measure the parathyroid hormone (PTH) levels. The volume and weight of the glands were calculated. The tissue was divided, with one aliquot being used for cell cycle analysis. The nuclei were acquired by flow cytometry and analyzed using CELLEIT software. Cell viability was assessed by flow cytometry and analyzed with LYSIS II software. Positive MIBI uptake was observed in 88.8% of patients. Focal MIBI uptake of one, two, or three glands was observed in 6, 11, and 8 patients, respectively. All patients experienced an 86% decrease in PTH blood level after SPTx compared to that before excision. A correlation was found between the volume of glands and the blood levels of intact PTH (iPTH) (r = 0.5, p < 0.05). A positive correlation was observed between MIBI uptake and the iPTH levels before SPTx (p < 0.01) and between the uptake of MIBI in the parathyroid glands and the cell cycle phases; low-grade uptake correlated with the G(0) phase and higher uptake with G(2)+S phase (r = 7, p < 0.01). No correlation was observed between MIBI uptake and the weight of the glands. MIBI scintigraphy accurately reflects the functional status of the hyperplastic parathyroid glands: Higher uptake grades correlated with the active growth phase. MIBI uptake does not reveal parathyroid enlargement; rather, it identifies the presence of hyperfunctioning autonomous glands. SPTx and total parathyroidectomy with autografting (TPTx+A) are the most widely accepted surgical approaches for patients with SPT. Reoperation for recurrence is necessary in 6% to 15% of cases. MIBI is now considered to be the radionuclide of reference for parathyroid gland scanning, although it is widely accepted that it produces poor results when trying to detect hyperplastic glands. PMID- 11038212 TI - Persistent hyperparathyroidism requiring surgical treatment after kidney transplantation. AB - There are not many publications describing long-term follow-up of persistent hyperparathyroidism requiring surgical treatment after kidney transplantation (PHSKT). In some patients adenomas, rather than multiglandular disease, have been incriminated as the cause of PHSKT. We reviewed the charts of 45 patients followed for 12 to 146 months (median 45 months) after parathyroidectomy for PHSKT. We compared them with (1) those of 951 patients receiving a kidney graft during the same period but not submitted to parathyroidectomy or (2) 90 matched controls selected from this cohort to determine the characteristics of PHSKT patients. The duration of pretransplant dialysis was significantly longer in PHSKT patients than in controls (5.78 +/- 0.41 vs. 3.41 +/- 0.24 years; p < 0.0001). A total of 166 glands were removed or biopsied. Except for one questionable case, no true adenoma was observed even when only one gland was enlarged. The outcome of surgery was not influenced by the technique (subtotal parathyroidectomy versus total parathyroidectomy and autografting) but depended on the amount of resected parathyroid tissue: no failures and 4 cases of hypoparathyroidism in 34 cases with no missing gland at cervical exploration; 3 failures and no permanent hypoparathyroidism in 11 cases with one or two missing glands. Excision of the enlarged glands only was sufficient to cure the patient. No recurrence was observed. Our results suggest that single gland enlargement in PHSKT results in most cases from different rates of involution of the parathyroids after successful kidney transplantation. When fewer than four glands are discovered, resection of all visible glands with or without grafting corrects hypercalcemia in more than 70% of the cases. PMID- 11038213 TI - Feasibility of sentinel lymph node biopsy and lymphatic mapping in nodular thyroid neoplasms. AB - Although the prognostic significance of occult lymph node metastases in thyroid cancer remains controversial, identifying these patients may help direct therapy. The purpose of this study was to determine the feasibility and safety of sentinel lymph node biopsy (SLNBx) in thyroid nodular disease. Patients undergoing thyroid resection, with no evidence of clinical lymphadenopathy, were enrolled. The nodule was injected with isosulfan blue vital dye. Blue-stained lymphatic channels were traced within the central compartment to the SLN, which was excised. A total of 40 patients underwent SLNBx; lymphatics were seen in 31 patients, and SLNs were found in 26. In 11 patients the lymphatic vessels were traced through the central compartment into the lateral or mediastinal compartments, although a central SLN was retrieved in only 6. Of the 18 patients with benign neoplasms, 14 had benign SLNs, and no SLN was found in 4. A thyroid lymphoma patient had a true positive SLN. In the 12 patients with papillary thyroid cancer (PTC), 6 had true positive SLNs, and 2 had a true negative SLN. In one patient with metastatic PTC, the parathyroid stained blue. Another patient with PTC had lateral lymphatic channels, but no SLN was found. There were two false negatives, proven by a node dissection in one and lateral uptake on (131)I scanning in the other. There were no postoperative complications. SLNBx for thyroid disease is feasible and safe. Potential staining of the parathyroids makes their identification before injection mandatory. The variable lymphatic drainage patterns and the two false-negative nodes indicate that further investigation is required before the procedure can be recommended for patients with thyroid disease. PMID- 11038214 TI - Method for dissection of mesenteric metastases in mid-gut carcinoid tumors. AB - With adequate medical management the midgut carcinoid tumor generally is an indolent malignancy associated with substantial life expectancy and appreciable life quality, even in the presence of liver metastases and significant tumor burden. Abdominal complications may occur in this entity of carcinoids owing to entrapment of intestines and encasement of mesenteric vessels by mesenteric metastases and associated marked mesenteric fibrosis. This may be the cause of abdominal pain, disabling diarrhea, weight loss to the extent of malnutrition, and eventually the risk of death with acute or chronic intestinal obstruction or intestinal gangrene. Operative removal of the mesentericointestinal lesion is often indicated to prevent or treat these complications but may be technically difficult when mesenteric metastases extend in the vicinity of major vessels in the mesenteric root. At laparotomy 56 patients with advanced midgut carcinoids underwent removal of the mesenteric tumor with a method for preserving the mesenteric vessels. This was feasible by mobilizing and releasing the right colon and mesenteric root from posterior adhesions, identifying the mesenteric artery below the pancreas, and free-dissecting this artery on the tumor capsule in the mobilized mesentery. Dissection was successful even with tumors initially judged inoperable unless tumor growth completely surrounded the mesenteric vessels or extended retroperitoneally. One patient was subjected to distal intestinal artery bypass. Symptom relief was been substantial and often of long duration after mesenteric tumor removal in patients who prior to surgery often had threatening intestinal ischemia. Patients with advanced midgut carcinoids may benefit markedly from dissectional removal of mesenteric tumors, which (conceivably better than conventional wedge resection) preserves the length of the remaining intestine. PMID- 11038215 TI - Familial papillary thyroid carcinoma: genetics, criteria for diagnosis, clinical features, and surgical treatment. AB - Hereditable predisposition to papillary thyroid carcinoma (PTC) and multinodular goiter (MNG) without evidence of an association with other malignancies as a distinct entity was recognized only recently. A meta-review of the literature on familial PTC (FPTC) was undertaken, and characteristics of families with frequent occurrence of PTC or MNG (or both) were summarized. A database on thyroid cancer patients maintained in our institution was searched for potential FPTC families. Clinical examinations were performed in 6 of 12 Hannover kindreds identified, and blood samples of all family members were collected for genetic analyses. Clinical presentations and histopathologic features of the FPTC cases were compiled. Based on the FPTC meta-review and own experience, predictive criteria to identify families at risk were developed: Exclusion criteria were previous radiation exposure and coincidence with neoplasia syndromes. Primary criteria for susceptibility to FPTC are (1) PTC in two or more first-degree relatives and (2) MNG in at least three first- or second-degree relatives of a PTC patient. Secondary criteria are diagnosis in a patient younger than 33 years, multifocal or bilateral PTC, organ-exceeding tumor growth (T4), metastasis (N1, M1), and familial accumulation of adolescent-onset thyroid disease. A hereditary predisposition to PTC is considered if both primary criteria or one primary criterion plus three secondary criteria are present. Family history-taking is recommended for all PTC patients to identify FPTC kindreds at risk. Blood relatives of FPTC index patients who harbor MNG should undergo thorough and regular clinical screening. Suspicious lesions should prompt early surgical intervention. PMID- 11038216 TI - Management of nonfunctioning islet cell carcinomas. AB - Tumors arising from the pancreatic islet cells are rare and represent a heterogeneous group of benign or malignant lesions. Most tumors present with well characterized syndromes, whereas others appear to be nonfunctioning. The clinical features of 11 men and 7 women with nonfunctioning islet cell carcinomas operated on between 1983 and 1998 were reviewed. The median patient age was 53.5 years (range 26-74 years). The most frequent presenting symptoms were abdominal pain (13 patients), weight loss (7 patients), and obstructive jaundice (4 patients). Gut hormone profiles were normal in all patients. Abdominal sonography and computed tomography localized the tumor in 17 patients, and correct prediction of an endocrine tumor was achieved in 12 patients. Six of seven patients showed a hypervascular tumor upon angiography, and seven of eight patients preoperatively had positive somatostatin receptor scintigraphy. At operation, regional or distant metastases were present in 15 (83%) and 6 (33%) patients, respectively. Eleven patients underwent potentially curative resections, and the remaining seven patients were managed palliatively by resection (four patients) or bypass procedures (three patients). Three patients had up to three more resection for metastases. Eight patients received postoperative octreotide, interferon alpha therapy, or both. The overall cumulative 5- and 10-year survival rates were 65.4% and 49.1%, respectively. Of the 11 patients who underwent curative resection, 10 were alive after a median follow-up of 63 months (range 7-180 months), but only 5 are free from disease. Although surgical cure is rare in nonfunctioning islet cell carcinomas, significant long-term palliation can be achieved in a large proportion of patients with an aggressive surgical approach and, when indicated, additional medical therapy. PMID- 11038217 TI - Curative resection of microgastrinomas based on the intraoperative secretin test. AB - The intravenous secretin injection test (secretin test) has been used for the differential diagnosis of gastrinoma. In this study we report that the intraoperative secretin test (IOS test) is also useful for determining the extent of curability in patients with Zollinger-Ellison syndrome (ZES). Twelve patients with ZES underwent surgical exploration and the IOS test. The results of the IOS test were obtained by rapid radioimmunoassay of the serum gastrin level (IRG) within 60 minutes. The test was diagnosed as negative when the maximum increase of serum IRG was less than 80 pg/ml and also less than 20% of the basal serum IRG level. Three of the twelve patients underwent pancreatoduodenectomy (PD), and two patients underwent distal pancreatectomy. Extirpation of duodenal tumors with dissection of regional lymph nodes was performed in seven patients. In two of the seven patients the IOS test remained positive after extirpation of the duodenal tumors and the dissection of regional lymph nodes. In one patient PD was performed on the basis of the positive results, and the IOS test became negative after PD. In the other patient, two tiny metastatic liver tumors were identified and were resected, but the IOS test did not become negative. We closed the abdomen in 11 patients when we obtained negative results from the IOS test. The results of the IOS test were almost identical to the data obtained by the standard assay postoperatively. The serum IRG levels of all but one patient fell to the normal level, and the secretin test became negative postoperatively. The IOS test is thus useful and indispensable for curative resection of microgastrinomas in patients with ZES. PMID- 11038218 TI - Gastrointestinal carcinoid tumors: long-term prognosis for surgically treated patients. AB - To evaluate long-term survival of patients with gastrointestinal carcinoid tumors and to assess factors that may influence prognosis, 154 patients (49% females, 51% males), median age 62 years (range 12-84 years) treated at our institution during 1972-1982 have been followed long term. Tumor location included the foregut (7%), midgut (62%), and hindgut (30%). Ninety-five percent of the patients underwent surgical or endoscopic excision of the primary tumor, with overall operative mortality and postoperative morbidity rates of 2. 6% and 11%, respectively. At follow-up, 60 patients (39%) were alive (median follow-up 18 years; range 1-26 years). The main causes of death included carcinoid tumor burden (32%), unrelated causes (45%), other malignancy (19%), and unknown causes (4%). Observed overall 5- and 10-year survivals were 69% and 53%, respectively. Survival was not related to gender or symptoms at presentation. However, age, embryologic origin, tumor size, depth of invasion, nodal status, and stage of disease proved to be of statistical significance (log-rank). In a multivariate Cox' model, only older age (> 62 years) [P = 0. 001, odds ratio (OR) = 3.4) and embryologic origin (midgut versus foregut) (P = 0.045, OR = 0.45) provided independent prognostic power when death from any cause was taken as the end point. This study confirms that patient's age and the site of the primary tumor have prognostic significance. Carcinoid tumors are neuroendocrine tumors with a relatively good prognosis, and long-term survival is possible despite advanced stages of disease. PMID- 11038219 TI - Are patients with multiple endocrine neoplasia type I prone to premature death? AB - Multiple endocrine neoplasia type I (MEN-I) is an autosomal dominant disorder characterized by endocrinopathies involving the anterior pituitary gland, parathyroid glands, and pancreas. The long-term prognosis for patients affected with this disorder is uncertain. To better characterize this prognosis, we performed a retrospective review of all patients with MEN-I treated at a single institution during the period 1951-1997. A group of 233 patients served as the study population. Their records were analyzed for confirmation of diagnosis, treatments received, long-term survival, and cause of death. Altogether, 108 eight male patients (46%) and 125 female patients (54%) were identified. At the conclusion of the study, 164 (70%) were alive and 69 (30%) were deceased, with a median follow-up for patients alive at last contact of 13.4 years (range < 1 month to 54.3 years). The cause of death was reliably obtained in 60 patients. Of these patients, 17 (28%) died of causes related to MEN-I, most commonly metastatic islet cell tumors (10 patients). The remaining patients died of causes unrelated to MEN-I, most commonly coronary artery disease and nonendocrine malignancies (14% each). The overall 20-year survival of MEN-I patients was 64% (95% CI was 56-72%), and that of an age- and gender-matched upper Midwest population was 81% (p < 0.001). Patients with MEN-I appear to be at increased risk of premature death. Earlier diagnosis and treatment of potentially malignant pancreatic islet cell neoplasms may result in a decrease of this premature mortality. PMID- 11038220 TI - Minimal incision parathyroidectomy: cure, cosmesis, and cost. AB - The goals of operative treatment of primary hyperparathyroidism are (1) cure; (2) minimal invasion; and (3) cost-effectiveness. The optimal strategy is controversial. Retrospective review of was undertaken 66 previously unoperated patients having minimal-incision, full-neck exploration by one surgeon over 29 months. A group of 51 women and 15 men had open full neck exploration under general anesthesia through a small (25-40 mm) incision using specifically selected instruments; patients remained hospitalized overnight. Preoperative sestamibi scans were obtained before referral for 17 patients: 11 had localized disease, and 6 did not (65% sensitivity). Four parathyroid glands were identified in 98% of patients; intraoperative frozen section was used selectively on a median of one gland per patient. About 76% of patients had single-gland disease, 6% had two-gland disease, and 18% had four-gland hyperplasia. One patient had four normal cervical parathyroid glands and an aortopulmonary window parathyroid adenoma resected at thoracotomy 1 week later; preoperative sestamibi scans failed to localize his disease. There were no nerve injuries and a 98% cure rate after initial cervical exploration. Excluding the cost of the sestamibi scans, there was no difference between those who had preoperative localization and those who did not; 60% of hospital costs were operating room time-related. Minimal-incision parathyroidectomy is effective for curing hyperparathyroidism and has excellent cosmetic results with negligible scar. Preoperative sestamibi scanning had no impact on cure or treatment costs. Strategies to improve cost-effectiveness must address the substantial costs of anesthesia and operating room services. PMID- 11038221 TI - Thyroid cancer in children of Ukraine after the Chernobyl accident. AB - The results of treatment of 330 children (< 14 years) and adolescents (15-18 years) with thyroid cancer who were operated on at the Institute of Endocrinology after the Chernobyl accident in 1986 were analyzed. The number of young patients increased after 1986 (1981-1985, 9 cases; 1986-1990, 37 cases; 1991-1995, 177 cases; 1996-1998, 116 cases). Most of these children and adolescents were younger than 8 years at the time of the accident (84.2%). More than half of the children (58.1%) lived in areas receiving the highest radiation exposure. These thyroid cancers developed after a short latent period, were more aggressive at presentation, and expressed regional (57.3%) or distant (14.5%) metastasis. Solid papillary cancers were present in 93.1%. Coexisting thyroid conditions were common (thyroid hyperplasia, 25.1%; nodular goiter, 18.8%; chronic thyroiditis, 10.2%). Most patients were treated by total thyroidectomy with intraoperative visualization of recurrent laryngeal nerves and parathyroid glands. When lymph node metastases were identified, a modified neck dissection was performed. Such operations were done in 277 (84.1%) patients. Postoperatively, the patients were treated with radioiodine and thyroid-stimulating hormone suppressive therapy. Postoperative complications included recurrent nerve palsy in 12.3% and permanent hypoparathyroidism in 6%. Operations for local recurrence of cancer were performed in 2.8% cases and for regional metastasis in 4%. The general mortality was 1. 8%. We anticipate that there will be more patients with thyroid cancer during the next few years. Therefore this high risk population for thyroid cancer must be carefully monitored and evaluated during the next several decades. PMID- 11038222 TI - Fundamental Concepts in Symbiotic Interactions: Light and Dark, Day and Night, Squid and Legume. AB - The legume-Rhizobium symbiosis and that between Euprymna scolopes and Vibrio fischeri show some surprising physiological similarities as well as differences. Both interactions rely on exchange of signal molecules, some of which are derived from bacterial cell surface molecules. Although the legume-Rhizobium symbiosis is nutritionally based as are many animal-microbe symbioses, it is not obligate because the plant initiates nodule formation only when the soil is deficient in nitrogen. In contrast, the squid-Vibrio symbiosis is obligate for the squid but is not nutritionally based. Rather, the bacteria produce light, which enables the animal to evade predators. These similarities and differences are described and discussed in term of the overall question of whether or not these two symbiotic relationships have evolved from commensal or pathogenic/parasitic interactions between prokaryotes and eukaryotes. PMID- 11038223 TI - An Introduction to the Biosynthesis of Chemicals Used in Plant-Microbe Communication. AB - Plants accumulate a diverse array of natural products, which can serve either to defend the plant against various microbes in its environment or to attract various microbes, both beneficial and pathogenic. Plants must also attract pollinators, repel or poison herbivores, compete with other plant species, and protect themselves from environmental dangers such as high light intensities. Some compounds have been implicated in playing a role in multiple interactions. Although the structures vary immensely in size and complexity, most are derived from a limited number of core biosynthetic pathways. This review briefly summarizes the biosynthetic origins of phenylpropanoid (including simple phenolics, flavonoids, anthocyanins and isoflavonoids), polyacetate, terpenoid, and alkaloid classes of metabolites. Compounds reported to be important in plant microbe, plant-animal, and plant-plant interactions will be given as examples of each of these classes. Other aspects of biosynthesis also will be discussed, including the timing or location of biosynthesis, the potential for genetic manipulation of these pathways, and various questions regarding the biosynthesis of these compounds. PMID- 11038224 TI - The Roles of Auxins and Cytokinins in Mycorrhizal Symbioses. AB - Most land plant species that have been examined exist naturally with a higher fungus living in and around their roots in a symbiotic partnership called a mycorrhiza. Several types of mycorrhizal symbiosis exist, defined by the host/partner combination and the morphology of the symbiotic structures. The arbuscular mycorrhiza (AM) is ancient and may have co-evolved with land plants. Emerging results from gene expression studies have suggested that subsets of AM genes were co-opted during the evolution of other biotrophic symbioses. Here we compare the roles of phytohormones in AM symbiosis and ectomycorrhizas (EC), a more recent symbiosis. To date, there is little evidence of physiologic overlap between the two symbioses with respect to phytohormone involvement. Research on AM has shown that cytokinin (CK) accumulation is specifically enhanced by symbiosis throughout the plant. We propose a pathway of events linking enhanced CK to development of the AM. Additional and proposed involvement of other phytohormones are also described. The role of auxin in EC symbiosis and recent research advances on the topic are reviewed. We have reflected the literature bias in reporting individual growth regulator effects. However, we consider that gradients and ratios of these molecules are more likely to be the causal agents of morphologic changes resulting from fungal associations. We expect that once the individual roles of these compounds are explained, the subtleties of their function will be more clearly addressed. PMID- 11038225 TI - Molecular Mechanisms in Root Nodule Development. AB - Under nitrogen-limiting conditions, bacteria from the family Rhizobiaceae establish a symbiosis with leguminous plants to form nitrogen-fixing root nodules. These organs require a coordinated control of the spatiotemporal expression of plant and bacterial genes during morphogenesis. Both plant and bacterial signals are involved in this regulation in the plant host. Plant genes induced during nodule development, the so-called nodulin genes, have been extensively characterized. Products of several of these genes show homologies to known regulators of signal transduction pathways in other plant or animal systems. Initial functional analysis of the molecular mechanisms implicated in nodulation have been undertaken using model legumes. Insertion mutagenesis and transgenic technologies to modify nodulin gene expression, as well as pharmacologic approaches, have been used to analyze molecular mechanisms involved in morphologic responses induced by the bacterial symbiont in the plant. G protein-mediated transduction mechanisms have been implicated, and the nin transcription factor appears to be required for early steps in nodule development. ENOD40, a gene coding for an RNA that contains only short ORFs, seems to be closely tied to nodule primordium formation. In addition, a vascular associated Kruppel-like transcription factor and small Rab type G-proteins affect bacteroid differentiation and the function of the nitrogen-fixing zone. These initial results presage a wealth of information that will be obtained from the application of genomic approaches to legumes. PMID- 11038226 TI - The Actinorhizal Symbiosis. AB - The term "actinorhiza" refers both to the filamentous bacteria Frankia, an actinomycete, and to the root location of nitrogen-fixing nodules. Actinorhizal plants are classified into four subclasses, eight families, and 25 genera comprising more than 220 species. Although ontogenically related to lateral roots, actinorhizal nodules are characterized by differentially expressed genes, supporting the idea of the uniqueness of this new organ. Two pathways for root infection have been described for compatible Frankia interactions: root hair infection or intercellular penetration. Molecular phylogeny groupings of host plants correlate with morphologic and anatomic features of actinorhizal nodules. Four clades of actinorhizal plants have been defined, whereas Frankia bacteria are classified into three major phylogenetic groups. Although the phylogenies of the symbionts are not fully congruent, a close relationship exists between plant and bacterial groups. A model for actinorhizal specificity is proposed that includes different levels or degrees of specificity of host-symbiont interactions, from fully compatible to incompatible. Intermediate, compatible, but delayed or limited interactions are also discussed. Actinorhizal plants undergo feedback regulation of symbiosis involving at least two different and consecutive signals that lead to a mechanism controlling root nodulation. These signals mediate the opening or closing of the window of susceptibility for infection and inhibit infection and nodule development in the growing root, independently of infection mechanism. The requirement for at least two molecular recognition steps in the development of actinorhizal symbioses is discussed. PMID- 11038227 TI - Plant Parasitic Nematodes: Habitats, Hormones, and Horizontally-Acquired Genes. AB - Plant parasitic nematodes are ubiquitous and cosmopolitan pathogens of vascular plants and exploit all parts of the roots and shoots, causing substantial crop damage. Nematodes deploy a broad spectrum of feeding strategies, ranging from simple grazing to the establishment of complex cellular structures (including galls) in host tissues. Various models of feeding site formation have been proposed, and a role for phytohormones has long been speculated, although whether they perform a primary or secondary function is unclear. On the basis of recent molecular evidence, we present several scenarios involving phytohormones in the induction of giant cells by root-knot nematode. The origin of parasitism by nematodes, including the acquisition of genes to synthesize or modulate phytohormones also is discussed, and models for horizontal gene transfer are presented. PMID- 11038228 TI - The Myriad Plant Responses to Herbivores. AB - Plant responses to herbivores are complex. Genes activated on herbivore attack are strongly correlated with the mode of herbivore feeding and the degree of tissue damage at the feeding site. Phloem-feeding whiteflies and aphids that produce little injury to plant foliage are perceived as pathogens and activate the salicylic acid (SA)-dependent and jasmonic acid (JA)/ethylene-dependent signaling pathways. Differential expression of plant genes in response to closely related insect species suggest that some elicitors generated by phloem-feeding insects are species-specific and are dependent on the herbivore's developmental stage. Other elicitors for defense-gene activation are likely to be more ubiquitous. Analogies to the pathogen-incompatible reactions are found. Chewing insects such as caterpillars and beetles and cell-content feeders such as mites and thrips cause more extensive tissue damage and activate wound-signaling pathways. Herbivore feeding is not equivalent to mechanical wounding. Wound responses are a part of the induced responses that accompany herbivore feeding. Herbivores induce direct defenses that interfere with herbivore feeding, growth and development, fecundity, and fertility. In addition, herbivores induce an array of volatiles that creates an indirect mechanism of defense. Volatile blends provide specific cues to attract herbivore parasites and predators to infested plants. The nature of the elicitors for volatile production is discussed. PMID- 11038229 TI - Signaling Organogenesis in Parasitic Angiosperms: Xenognosin Generation, Perception, and Response. AB - Parasitic strategies within the angiosperms generally succeed by tightly coupling developmental transitions with host recognition signals in a process referred to as xenognosis. Within the Scrophulariaceae, Striga asiatica is among the most studied and best understood parasitic member with respect to the processes of host recognition. Specific xenognosins regulate seed germination, the development of the host attachment organ, the haustorium, and several later stages of host parasite integration. Here we discuss the signals regulating the development of the haustorium, the critical vegetative/parasitic transition in the life cycle of this obligate parasite. We provide evidence for the localized production of H(2)O(2) at the Striga root tip and suggest how this oxidant is used to exploit host peroxidases and cell wall pectins to generate a simple benzoquinone signal. This benzoquinone xenognosin proves to be both necessary and sufficient for haustorial induction in cultured seedlings. Furthermore, evidence is provided that benzoquinone binding to a redox active site completes a "redox circuit" to mediate signal perception. This redox reaction regulates the time-dependent expression of specific marker genes critical for the development of the mature host attachment organ. These studies extend the emerging series of events necessary for the molecular regulation of organogenesis within the parasitic plants and suggest novel signaling features and molecular mechanisms that may be common across higher plants. PMID- 11038230 TI - Role of Phenylurea Cytokinin CPPU in Apical Dominance Release in In Vitro Cultured Rosa hybrida L. AB - The effect of purine (BA) and phenylurea (CPPU) cytokinins on apical dominance release in in vitro cultured Rosa hybrida L., cv. Madelon and Motrea was evaluated. Cv. Madelon shows stronger natural apical growth and fewer branches than cv. Motrea in vivo and in vitro. We examined the effects under three conditions, without the addition of the auxin IBA, in the presence of IBA, and in material pretreated with a pulse of IBA. Results were scored weekly for 4 weeks. BA and CPPU stimulated axillary bud break, and higher numbers of open buds were recorded in the presence of CPPU. When CPPU cytokinin was added to culture medium, physiologic features such as bud sprouting and shoot fresh and dry weight were enhanced. CPPU was also highly efficient for overcoming IBA inhibition of bud outgrowth. Different cultivar responses were observed. PMID- 11038231 TI - Putrescine and Silver Nitrate Influences Shoot Multiplication, In Vitro Flowering and Endogenous Titers of Polyamines in Cichorium intybus L. cv. Lucknow Local. AB - The influence of putrescine (Put) and AgNO(3) on shoot multiplication, in vitro flowering and endogenous titers of polyamines in Cichorium intybus L. cv. Lucknow local was investigated. Exogenous administration of Put at a concentration of 40 mM resulted in maximum tissue response in terms of shoot numbers (34.6 +/- 2.61) and shoot lengths (7.6 +/- 0.57 cm) on MS media supplemented with 2-iP (2.0 mg L( 1)) and GA(3) (0.5 mg L(-1)) as observed on the 35(th) day. Exogenous application of 40 uM AgNO(3) resulted in maximum shoot number (36.8 +/- 2.63) and shoot lengths (7.9 +/- 0.76 cm) on day 35 on the same media. Endogenous titers of conjugated spermidine decreased sharply from day 7-21, whereas endogenous conjugated spermine levels peaked on day 28 (1265 +/- 94.9 nmoles g(-1) FW), after treatment with 40 mM Put. Whereas, AgNO(3) (40 uM) fed samples resulted in higher titers of endogenous conjugated spermine (1405 +/- 105.6 nmoles g(-1) FW, 3.62 fold over control) on day 14. All other treatments showed decreasing endogenous levels during the whole culture period. Both Put (40 mM) and AgNO(3) (40 uM) resulted in floral initiation and floral development on day 28 and 14 (3.76 +/- 0.16, 4.2 +/- 0.21 flowers per shoot apices), respectively. To investigate the role of Put (40 mM) and AgNO(3) (40 uM) on morphogenetic response and endogenous conjugated polyamine titers in shoots of chicory, polyamine inhibitors (DFMA and DFMO) were used. The morphogenetic response and the endogenous conjugated pool of polyamines were diminished in DFMA and DFMO treatments, but could be restored by addition of Put (40 mM) and AgNO(3) (40 uM). Under exogenous Put feeding, ethylene production was reduced in shoot cultures of chicory. This study shows for the first time the influence of polyamines on multiple shoot initiation from axillary buds of C. intybus L. cv. Lucknow local and also indicates the promotive effect of Put and AgNO(3) on autoregulation of polyamine biosynthesis, thereby regulating in vitro flowering, the endogenous pool of polyamines and shoot multiplication. PMID- 11038232 TI - Heating of food and haemoglobin adducts from carcinogens: possible precursor role of glycidol. AB - Studies of adducts from reactive compounds to haemoglobin (Hb) by gas chromatography-tandem mass spectrometry according to the N-alkyl Edman method reveals the occurrence of N-(2,3-dihydroxypropyl)valine (diHOPrVal) at levels of 1-2 pmol/g Hb, in persons without known exposure. The hypothesis that this background originates from glycidol or related compounds during heating of food was tested in experiments with rats. Animals fed fried animal feed for 30 or 72 days showed an increase of the diHOPrVal level by about 50% compared with controls. Several arguments, such as the formation of reactive oxiranes by heat induced dehydration of glycol configurations in glycerol and sugars, support the idea that glycidol (or e.g. glycidyl esters) are precursors of the adduct. In Hb samples, reduced for stabilisation of aldehyde adducts, relatively high levels of adducts determined as diHOPrVal were found, although without significant relation to frying of the feed. There is thus no indication that reduction in vivo of, for example, the Schiff base from glyceraldehyde, is a pathway for formation of the diHOPrVal. The background level of diHOPrVal in humans Hb is low, and the cancer risk associated with exposure to the specific alkylator-probably glycidol-formed in cooking, is therefore presumably low. The result implies, however, that low molecular mass mutagenic oxiranes formed during the heating of food should be studied further. PMID- 11038233 TI - In vitro genotoxicity testing of ARASCO and DHASCO oils. AB - ARASCO and DHASCO oils are microbially-derived triglycerides rich in arachidonic (20:4n-6) and docosahexaenoic (22:6n-3) acids, respectively. Both oils were tested for mutagenic activity in three different in vitro mutagenesis assays. All assays were conducted with and without metabolic activation. Neither ARASCO nor DHASCO oil was mutagenic in the Ames reverse mutation assay using five different Salmonella histidine auxotroph tester strains, nor were the oils mutagenic in the mouse lymphoma TK(+/-) forward mutation assay. The oils showed no clastogenic activity in chromosomal aberration assays performed with Chinese hamster ovary cells. Based on these assays, neither ARASCO nor DHASCO oils appear to have any genotoxic potential. PMID- 11038234 TI - Effects of butylated hydroxyanisole and butylated hydroxytoluene on DNA adduct formation and arylamines N-acetyltransferase activity in PC-3 cells (human prostate tumor) in vitro. AB - The effects of butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) on the N-acetyltransferase (NAT) activity and DNA adduct formation in PC-3 cells (human prostate tumor) was studied. PC-3 cells were placed into tissue culture flasks and grown in an incubator as cytosols and intact cells. The BHA or BHT were added to the cytosols and intact cells. The NAT activity in cytosol and intact PC-3 cells were measured by HPLC assaying exhibited for the amounts of N acetyl-2-aminofluorene and N-acetyl-p-aminobenzoic acid, 2-aminofluorene and p aminobenzoic acid. The NAT activity in PC-3 cells and cytosols were inhibited by BHA or BHT in a dose-dependent manner; that is, the higher the concentrations of BHA or BHT the higher inhibition of NAT activity. The NAT values of K(m) and V(max) from PC-3 cells were also decreased by BHA or BHT in both cytosols and intact cells. The data also demonstrated concomitant exposure to BHA or BHT decreased AF-DNA adduct formation which was seen in the PC-3 cells. In addition, the formation of DNA adduct was decreased after BHA or BHT exposure. These findings suggested the usefulness of using human cultured PC-3 cells for assessing arylamine-induced DNA adduct formation. Furthermore, the findings illustrate how effectively BHA or BHT reduce the adduct formation. PMID- 11038235 TI - Toxicological evaluation of New Zealand deer velvet powder. Part I: acute and subchronic oral toxicity studies in rats. AB - Potential toxic effects of acute and subchronic dosage regimens of deer velvet powder have been assessed in rats following OECD guidelines. In the acute study, rats of both sexes were exposed to a single dose of 2 g/kg body weight. There was no mortality or other signs of toxicity during 14 days' observation. Furthermore, no significant alteration either in relative organ weights or their histology was discernible at terminal autopsy. In the 90-day subchronic study, deer velvet was administered in 1 g/kg daily doses by gavage to rats. A control group of rats received water only. There was no effect on body weight, food consumption, clinical signs, haematology and most parameters of blood chemistry including carbohydrate metabolism, liver and kidney function. No significant differences were seen between the mean organ weights of the adrenal, kidney and brain in rats treated with deer velvet and control rats. However, there was a significant difference (P<0.05) in the group mean relative liver weight (3.52 +/- 0.30 vs 3.81 +/- 0.26 g/100 g body weight) of deer velvet-treated and control male rats. The gross necropsy and pathological examination of rats treated with deer velvet did not reveal any abnormalities in tissue morphology. Based on these results, it may be concluded that rats had no deer velvet treatment-related toxicological and histopathological abnormalities at the doses administered, despite the observed minor changes in liver weight. PMID- 11038236 TI - Inhibition by curcumin of diethylnitrosamine-induced hepatic hyperplasia, inflammation, cellular gene products and cell-cycle-related proteins in rats. AB - Curcumin (CCM), a major yellow pigment of turmeric obtained from powdered rhizomes of the plant Curcuma longa Linn, is commonly used as coloring agent in foods, drugs and cosmetics. In this study we report that gavage administration of 200 mg/kg or 600 mg/kg CCM effectively suppressed diethylnitrosamine (DEN) induced liver inflammation and hyperplasia in rats, as evidenced by histopathological examination. Immunoblotting analysis showed that CCM strongly inhibited DEN-mediated the increased expression of oncogenic p21(ras) and p53 proteins in liver tissues of rats. In cell-cycle-related proteins, CCM selectively reduced the expression of proliferating cell nuclear antigen (PCNA), cyclin E and p34(cdc2), but not Cdk2 or cyclin D1. Moreover, CCM also inhibited the DEN-induced increase of transcriptional factor NF-kappa B. However, CCM failed to affect DEN-induced c-Jun and c-Fos expression. It has become widely recognized that the development of human hepatocellular carcinoma (HCC) is predominantly due to the chronic inflammation by virus, bacteria or chemical. Our results suggest a potential role for CCM in the prevention of HCC. PMID- 11038237 TI - Sex- and organ-specific toxicity in normal and malnourished rats fed thermoxidized palm oil. AB - The effects of free radical toxicity as induced by chronic consumption of thermoxidized palm oil (TPO) diet on organ size of normal animals, their first filial offspring and malnourished rats, were studied. Tissue- and sex-specific toxicity was revealed. The TPO diet significantly (P<0.01) reduced lung and kidney mass in normal male rats but female rats remained unaffected. Hearts of first filial offspring of both male and female rats were, however, enlarged while lung, liver and kidneys of first filial female offspring were additionally reduced in size (P<0.01). This information suggests that the observed toxicities could be cumulative for female offspring. Malnutrition protected against toxic injury because none of the kwashiorkoric animals rehabilitated on the toxic diet showed any overt symptoms of toxicity. PMID- 11038238 TI - Effects of the seafood toxin domoic acid on glutamate uptake by rat astrocytes. AB - Pronounced glutamic acid uptake was observed after only 15 min with glutamate concentrations of 60 nmol/mg protein when astrocytes were incubated with 1 mM glutamic acid. The uptake increased with time to a steady-state glutamate level of above 160 nmol/mg protein by 45 min. The uptake was energy dependent. Reduced temperature (0 degrees C) and ouabain (100 microM) inhibited uptake by 86.7% (P<0.001; n=18) and 84.4% (P<0.001; n=18), respectively, when compared with controls. After exposure of astrocytes to glutamate (1 mM) in the incubation medium, in the presence of domoic acid (10 and 100 microM) at 5 and 60 min, domoic acid (10 microM) elevated glutamate uptake by 64.0% (P<0.05; n=34) at 5 min but decreased glutamate uptake by 47.8% (P<0.01; n=19) at 60 min compared with controls. A higher dose of domoic acid (100 microM) decreased glutamate uptake by 49.6% (P<0.01; n=20) and 61.3% (P<0.001; n=20) at 5 and 60 min, respectively, compared with controls. This study suggests that domoic acid may induce neurotoxicity because of the failure of astrocytes to remove extracellular glutamate. This may contribute to excitotoxic injury. PMID- 11038239 TI - Excretion of volatile nitrosamines in a rural population in relation to food and drinking water consumption. AB - Urinary excretion of volatile nitrosamines was assessed in 59 non-smokers living in a rural county of Quebec, Canada. Water and food intakes were measured by means of a 24-hour recall. Nitrates were analyzed in the tap water of all participants (geometric mean=2.0 mg nitrate-N/L) and dietary intakes of nitrate and vitamins C and E were estimated via a validated Canadian food database. Urine was collected over the same 24-hour period and analyzed for nitrates by hydrazine reduction and for volatile nitrosamines by gas-chromatography/mass spectrometry. N-Nitrosopiperidine (NPIP) was found in urine samples from 52 of the 59 subjects. Geometric mean of NPIP urinary excretion was 67 ng/day and maximum value was 1045 ng/day. No other volatile nitrosamine was detected. There was a correlation between urinary nitrate excretion and total nitrate intake (r=0.71, P < 0.001). However, no relationship was found between urinary NPIP excretion and either nitrate excretion, dietary or water nitrate intakes. NPIP excretion was significantly correlated to coffee intake (r=0.40, P=0.002) and this relation was not modified by vitamin intake. We conclude that nitrate intake is not related to nitrosamine excretion in this rural population. The influence of coffee consumption on NPIP excretion deserves further attention. PMID- 11038240 TI - Assessment of the carcinogenicity associated with oral exposures to hydrogen peroxide. AB - Concern regarding hydrogen peroxide (H(2)O(2)) carcinogenicity arises from its ability to act as a strong oxidizing agent. In short-term genotoxicity tests, H(2)O(2) has given predominantly positive results; however, these assays have been performed using either bacterial strains engineered to be exquisitely sensitive to oxidant damage, or mammalian cells deficient in antioxidant enzymes. Significantly, the addition of antioxidant protective measures (normally present in vivo) to these assay systems protects against H(2)O(2) genotoxicity. In most whole animal studies, H(2)O(2) exposure neither initiates nor promotes tumors. In mice, however, 0.4% H(2)O(2) in drinking water was reported to induce hyperplastic lesions of the duodenum and to erode areas in the glandular stomach epithelium. Owing to the chemistry of dilute H(2)O(2) solutions and the anatomy/physiology of the gastrointestinal tract, it is unlikely that orally ingested H(2)O(2) reaches the duodenum. Instead, greatly decreased water consumption and the resultant abrasion of the luminal lining on ingestion of pelleted dry rodent chow is the most likely cause of the observed gastric and duodenal lesions following H(2)O(2) administration in drinking water. Significantly, when hamsters received high doses of H(2)O(2) by gastric intubation (and water intake was not affected), the gastric and duodenal epithelia appeared normal. In-depth analysis of the available data supports the conclusion that oral ingestion of H(2)O(2) should not be considered a carcinogenic threat. PMID- 11038241 TI - Absorption, distribution, metabolism and excretion of intravenously and dermally administered triethanolamine in mice. AB - Triethanolamine (TEA) is an amino alcohol having widespread applications in consumer goods and as an industrial chemical. A number of relatively high-dose dermal toxicity studies have been conducted in rats and mice reflecting the principal route of human exposure to TEA. The absorption, distribution, metabolism and excretion (ADME) of (14)C-TEA derived radioactivity were determined in male C3H/HeJ mice following dermal application of 2000 mg/kg (neat) or, to characterize blood kinetics, intravenous (iv) injection of 1 mg/kg (14)C TEA. Balance and excretion data were also collected in mice utilizing several dermal dosing scenarios (1000 mg/kg in acetone, 2000 mg/kg neat, 2000 mg/kg in water) and, for comparative purposes, in male Fischer 344 rats dosed dermally with 1000 mg/kg neat (14)C-TEA. Urine, feces, expired CO(2) (iv) and, where appropriate, blood were collected over a 24- or 48-hour period post-dosing. The half-life for dermal absorption of radioactivity was estimated to be 1.3 hours. Intravenously administered radioactivity was eliminated in a biphasic manner with a prominent initial phase (half-life of 0.3 hr) followed by a slower terminal phase (half-life of 10 hr). Radioactivity was excreted primarily via the urine (49-69%) as unmetabolized TEA, regardless of dosage, route or vehicle used. Fecal excretion of radioactivity comprised 16-28% of dose administered. The body burden at sacrifice (sum of liver, kidney, carcass and non-application site skin) ranged from 3 to 6% of the dose. It was concluded that TEA is absorbed extensively following dermal application to mice at dosages relevant to toxicity testing and that acetone or water vehicles do not appear to significantly alter total uptake. Significantly, the blood kinetics and ADME of TEA in mice and/or rats differs from that of a related chemical, diethanolamine, which appears to be more toxic to rodents than TEA. PMID- 11038242 TI - Consequences of Aroclor 1254 ingestion on the menstrual cycle of rhesus (Macaca mulatta) monkeys. AB - A group of 80 female rhesus (Macaca mulatta) monkeys were randomly distributed to four similar test rooms (20 monkeys/room) and then randomly allocated to one of five test groups (four females/test group/room). The objective of the study was to ascertain the toxicological and reproductive effects of Aroclor 1254 ingestion at dose levels of 0, 5, 20, 40 or 80 microg Aroclor 1254/kg body weight per day (Arnold et al., 1993a,b, 1995, 1996, 1997). It was deemed necessary to establish the menstrual patterns for all the monkeys both before and after the start of dosing so as to provide an appropriate baseline from which potential treatment effects could be ascertained. The data presented herein were obtained during the first 3 years after the start of dosing, or the study's pre-mating phase. At the end of the first 2 years of dosing, the monkeys attained a qualitative pharmacokinetic steady state regarding the levels of polychlorinated biphenyls in their adipose tissue. Upon termination of the study, a number of monkeys were found to have endometriosis, adenomyosis or uterine leiomyomas (Arnold et al., 1996, 1997). These monkeys were designated as having gynecological abnormalities which were considered to be a factor in the analysis of the menstrual data. The menstrual data (i.e. menses frequency, cycle length and menses duration) were subjected to a statistical assessment to see whether year, quarter, gynecological abnormalities or dose of Aroclor 1254 had any effect on menses frequency, menstrual cycle length (i.e. the first day of menses until the day prior to the start of the next menses) and/or menses duration (i.e. the number of days of haemorrhagic discharge). The only consistent statistically significant effect found was that gynecological abnormalities increased menses duration (P<0.05) in all 12 quarters of the premating observation period. This effect was significant during both the pre- (P=0.0004) and post- (P< or =0.0001) pharmacokinetic steady state intervals. While there was some indication of seasonality regarding menstrual cycle length and menses duration when these data were compared on a quarterly basis during the first 2 years of the study (P=0.043; P< or =0.0001, respectively), this effect was not evident during the third year (P=0.21; P=0.31, respectively). In particular, the effect of quarter on menses cycle length was most evident during the first year, with the shortest cycles occurring during the first or spring quarter and the longest in the third or fall quarter. However, menses duration was shortest in the first quarter during the first 2 years and tended to peak in the second quarter of all 3 years, while generally diminishing in the third and fourth quarters. There was also an increase in menses duration with increasing time on test for all groups. In addition, Aroclor 1254 treatment appeared to have some effect on menses duration when menses duration was plotted against dose group, but the effect was not statistically significant (P>0. 05). It was concluded that the ingestion of Aroclor 1254 at dose levels up to 80 microg/kg body weight/day by rhesus monkeys did not have any appreciable biological effect on menstrual frequency, menstrual cycle length or menses duration. However, gynecological abnormalities significantly increased menses duration during the three-year observation period. PMID- 11038243 TI - Study of adjuvant effect of model surfactants from the groups of alkyl sulfates, alkylbenzene sulfonates, alcohol ethoxylates and soaps. AB - The sodium salts of representatives of anionic surfactants, dodecylbenzene sulfonate (SDBS), dodecyl sulfate (SDS) and coconut oil fatty acids, and a nonionic surfactant, dodecyl alcohol ethoxylate, were studied for adjuvant effect on the production of specific IgE antibodies in mice. The surfactants were injected subcutaneously (sc) in concentrations of 1000, 100, 10 or 1 mg/l, respectively, together with 1 microg of ovalbumin (OVA). In addition, groups of mice received OVA in saline (control group) or in Al(OH)(3) (positive adjuvant control group). After the primary immunization the mice were boosted up to three times with OVA (0.1 microg sc) in saline. OVA-specific IgE antibodies were determined by the heterologous mouse rat passive cutaneous anaphylaxis test. The results were confirmed by a specific ELISA method. After the first booster, the Al(OH)(3) group and the 10 mg/l SDS group showed a statistically significant increase in OVA specific IgE levels. After two boosters, a statistically significant suppression in OVA-specific IgE production occurred with SDS (1000 mg/l), SDBS (1000 and 100 mg/l), coconut soap (1000 mg/l) and the alcohol ethoxylate (10 mg/l). This study suggests that a limited number of surfactants possess an adjuvant effect whereas all surfactants at certain levels can suppress specific IgE production. PMID- 11038244 TI - A caspase-3-like protease is involved in NF-kappaB activation induced by stimulation of N-methyl-D-aspartate receptors in rat striatum. AB - Glutamate receptor stimulation reportedly activates NF-kappaB in vitro and in vivo, although underlying mechanisms remain to be elucidated. Here we evaluated the role of proteases in mediating N-methyl-D-aspartate (NMDA) receptor agonist induced NF-kappaB activation and apoptosis in rat striatum. The intrastriatal infusion of quinolinic acid (QA, 60 nmol) had no effect on levels of NF-kappaB family proteins, including p65, p50, p52, c-Rel and Rel B. In contrast, QA decreased IkappaB-alpha protein levels by 60% (P<0. 05); other members of the IkappaB family, including IkappaB-beta, IkappaB-gamma, IkappaB-epsilon and Bcl-3, were not altered. The QA-stimulated degradation of IkappaB-alpha was completely blocked by the NMDA receptor antagonist MK-801. QA-induced IkappaB-alpha degradation and NF-kappaB activation were not affected by the proteasome inhibitor MG-132 (1-4 microg). On the other hand, the caspase-3 inhibitor Ac DEVD.CHO (2-8 microgram) blocked QA-induced IkappaB-alpha degradation in a dose dependent manner (P<0.05). Ac-DEVD.CHO (4 microgram) also substantially reduced QA-induced NF-kappaB activation (P<0.05), but had no effect on QA-induced AP-1 activation. Furthermore, Ac-DEVD.CHO, but not MG-132, dose-dependently attenuated QA-induced internucleosomal DNA fragmentation. These findings suggest that NF kappaB activation by NMDA receptor stimulation involves IkappaB-alpha degradation by a caspase-3-like cysteine protease dependent mechanism. Caspase-3 thus appears to contribute to the excitotoxin-induced apoptosis in rat striatal neurons occurring at least partially as a consequence of NF-kappaB activation. PMID- 11038245 TI - Effects of post-mortem delay on subunits of ionotropic glutamate receptors in human brain. AB - The effect of post-mortem delay on the stability of the protein subunits that combine to form NMDA and AMPA type glutamate receptors has been assessed in samples of human brain tissue. While most of the subunits (i.e. GluR1, GluR2/3, GluR4, NR1) appear to be stable for up to 18 h post-mortem, the NR2A and NR2B subunits appear to be proteolyzed rapidly following death. These results are consistent with the concept that the proteolytic products of NR2A and NR2B, although at smaller molecular sizes than the full-length protein, are all identifiable on Western blots. Thus, a method is proposed that allows for the estimation of the levels of these labile proteins even in samples obtained up to 18 h post-mortem. Using this method we have estimated the levels of all AMPA and NMDA receptor subunits in selected (i.e. hippocampus, frontal and entorhinal cortex) brain tissue samples obtained from control patients and patients who have died with Alzheimer's disease. Modest decreases in NMDA receptor subunits NR1, NR2A, and NR2B were found in the hippocampus and in frontal cortex while little or no change in any of these subunits were documented in entorhinal cortex. Subunits for AMPA receptors (GluR1, GluR2/3, and GluR4) appeared to show a generalized decrease in all these tissues. As a surrogate marker for overall decreases due to generalized neuronal cell death, levels of neuron-specific enolase were measured in all tissues and were found to be nearly identical in control and Alzheimer's brains. PMID- 11038246 TI - Mechanism for increase in expression of cerebral diazepam binding inhibitor mRNA by nicotine: involvement of L-type voltage-dependent calcium channels. AB - We investigated the mechanisms underlying the increase in diazepam binding inhibitor (DBI) and its mRNA expression induced by nicotine (0.1 microM) exposure for 24 h using mouse cerebral cortical neurons in primary culture. Nicotine induced (0.1 microM) increases in DBI mRNA expression were abolished by hexamethonium, a nicotinic acetylcholine (nACh) receptor antagonist. Agents that stabilize the neuronal membrane, including tetrodotoxin (TTX), procainamide (a Na(+) channel inhibitor), and local anesthetics (dibucaine and lidocaine), dose dependently inhibited the increased expression of DBI mRNA by nicotine. The nicotine-induced increase in DBI mRNA expression was inhibited by L-type voltage dependent Ca(2+) channel (VDCC) inhibitors such as verapamil, calmodulin antagonist (W-7), and Ca(2+)/calmodulin-dependent protein kinase II (CAM II kinase) inhibitor (KN-62), whereas P/Q- and N-type VDCC inhibitors showed no effects. In addition, nicotine exposure for 24 h induced [3H]nicotine binding to the particulate fractions of the neurons with an increased B(max) value and no changes in K(d). Under these conditions, the 30 mM KCl- and nicotine-induced 45Ca(2+) influx into the nicotine-treated neurons was significantly higher than those into non-treated neurons. These results suggest that the nicotine stimulated increase in DBI mRNA expression is mediated by CAM II kinase activation resulting from the increase in intracellular Ca(2+) through L-type VDCCs subsequent to the neuronal membrane depolarization associated with nACh receptor activation. PMID- 11038247 TI - Stress regulation of adrenocorticosteroid receptor gene transcription and mRNA expression in rat hippocampus: time-course analysis. AB - Neuronal glucocorticoid receptor (GR) and mineralocorticoid receptor (MR) proteins mediate the transcriptional effects of circulating glucocorticoids. These receptors bind the same DNA response element, yet mediate quite different cellular functions. The present study tests the hypothesis that acute and chronic stress, which cause increases in glucocorticoids sufficient to bind the GR, will regulate expression of the GR and MR genes in the hippocampal formation. Analysis of MR gene transcription using an intronic MR probe revealed a transient 50% decrease in MR hnRNA in CA1, CA3 and dentate gyrus at 60-120 min post-stress, consistent with glucocorticoid down-regulation of the MR gene. However, no changes were seen in full-length MR mRNA at any post-stress time point. In contrast, GR hnRNA was not affected by acute stress, but GR mRNA was decreased 120 min post stress in all hippocampal subregions. Chronic stress exposure down regulated GR mRNA in CA3 only; effects were first evident 7 days post stress and persisted for the entire stress time-course (28 days). There was no evidence for down-regulation of GR hnRNA or MR hnRNA/mRNA at any point in the chronic stress regimen. The transient decrease in MR hnRNA in the absence of mRNA changes suggests increased MR mRNA stability. In contrast, acute stress decreases the availability of GR mRNA without demonstrably affecting transcription, suggesting reduced GR mRNA stability. The results suggest that acute stress alters GR mRNA expression by largely post-transcriptional mechanisms. However, elevations in basal corticosterone secretion seen following chronic stress are not sufficient to markedly down-regulate GR/MR expression in a long-term fashion. PMID- 11038248 TI - Induction of NGFI-B mRNA following contextual fear conditioning and its blockade by diazepam. AB - Expression of the immediate-early gene, NGFI-B (nerve growth factor inducible gene B), was examined in the amygdala, hippocampus, and neocortex following contextual fear conditioning. Rats were either handled, placed within the testing context without receiving the footshock, received a footshock immediately upon placement within the context, or received a footshock after a 3-min delay (delayed-shock). Only the delayed-shock group displayed a fear response (freezing) in the post-shock period and in a retention test 24 h after fear conditioning. Expression of NGFI-B mRNA was increased in the dorsolateral part of the lateral nucleus of the amygdala (LaDL) and the neocortex 30 min following conditioning in the delayed-shock group compared to the other three groups. In addition, following a retention test conducted 24 h after fear conditioning, NGFI B mRNA expression was increased in the neocortex of the delayed-shock group compared to the handled group. In a subsequent experiment, the effects of pretreatment with the anxiolytic drug, diazepam, on fear conditioning and the concomitant increases in NGFI-B mRNA were investigated. Rats administered a 2.5 mg/kg, i.p. dose of diazepam before fear conditioning did not acquire contextual fear as demonstrated by a lack of freezing in a retention test. Although diazepam blocked fear conditioning while the 40% propylene glycol, 10% ethanol vehicle solution did not, both diazepam and the vehicle reduced the conditioning-induced increase in NGFI-B expression in the LaDL. In contrast, the fear-conditioning induced NGFI-B increase in the neocortex was blocked by diazepam, but not by the vehicle. The data suggest that the transcriptional factor NGFI-B in the LaDL and neocortex may play a functional role in learning and memory of contextual fear, but blocking the increase in NGFI-B expression in the LaDL is not essential for diazepam to interfere with fear conditioning. PMID- 11038250 TI - Defective extracellular calcium (Ca(o))-sensing receptor (CaR)-mediated stimulation of a Ca(2+)-activated potassium channel in glioblastoma cells transfected with a dominant negative CaR. AB - Glioblastoma cells exhibit several forms of sensitivity to extracellular calcium (Ca(o)) that might be conferred by the Ca(o)-sensing receptor (CaR) that is intimately involved in the maintenance of Ca(o) homeostasis by various cell types. This receptor is expressed in human glioblastoma cell line, U87, and here we show that CaR activators stimulate a Ca(2+)-activated potassium (K(+)) channel (CAKC) with a conductance of 140 pS. The responses to CaR activators, however, were blunted in U87 cells transfected with a CaR bearing an inactivating mutation (R185Q) that has previously been shown to exert a dominant negative (DN) action on the wild type receptor. Raising Ca(o) from 0.75 to 2.0 mM or addition of a polycationic CaR agonist, each activated CAKC in nontransfected wild type and empty vector-transfected U87 cells, while they had little or no effect on channel activity in cells expressing the DN CaR (DN-CaR cells). In nontransfected wild type and empty vector-transfected cells, the specific 'calcimimetic' CaR activator, NPS R-467, stimulated the channel, while its less active stereoisomer, NPS S-467, did not. In DN-CaR cells, in contrast, NPS R-467, had no effect on channel activity, suggesting defective coupling of the CaR to this ion channel. CaR-mediated stimulation of these K(+) channels could lead to membrane repolarization and related changes in cellular function under normal conditions. Since the R185Q mutation in the CaR produces a more severe phenotype in humans than most inactivating mutations of this receptor, some of its clinical consequences could potentially result from abnormal CaR-dependent channel functioning. PMID- 11038249 TI - Pharmacological profiles of selective non-peptidic delta opioid receptor ligands. AB - Several non-peptidic opioids have been synthesized recently as part of a program to develop selective delta receptor agonists. In this study, the affinities of a set of compounds for cloned delta and mu opioid receptors expressed in HEK 293 cell lines were determined by competition analysis of [3H]bremazocine binding to membrane preparations. All compounds studied exhibited high affinity and selectivity, with apparent dissociation constants in the range of 0.6-1.7 nM for the delta opioid receptor and 240-1165 nM for the mu opioid receptor. We next sought to determine which domain of the delta receptor was critical for mediating the highly selective binding by analysis of ligand affinities for mu/delta receptor chimeras. Receptor binding profiles suggested that a critical site of receptor/ligand interaction was located between transmembrane domain 5 (TM5) and TM7 of the delta receptor. Substitution of tryptophan 284, located at the extracellular surface of TM6, with lysine, which is found at the equivalent position in the mu opioid receptor, led to a spectrum of effects on affinities, depending on the ligand tested. Affinities of SB 219825 and SB 222941 were particularly sensitive to the substitution, displaying a 50-fold and 70-fold decrease in affinity, respectively. Activities of the delta receptor-selective agonists were tested in two functional assays. Brief exposure of HEK 293 cells expressing delta opioid receptors with selective ligands induced phosphorylation of MAP kinase, although the non-peptidic ligands were less efficacious than the enkephalin derivative DADL (Tyr-D-Ala-Gly-Phe-D-Leu). Similarly, chronic exposure of HEK 293 cells expressing delta opioid receptors with selective, non-peptidic ligands, with the exception of SB 206848, caused receptor down-regulation, however, the SB compounds were less efficacious than DADL. PMID- 11038251 TI - Spatiotemporal expression of BDNF in the hippocampus induced by the continuous intracerebroventricular infusion of beta-amyloid in rats. AB - The beta-amyloid protein (Abeta) is the major component of senile plaques found in the brain in Alzheimer's disease (AD). Its neurotoxic properties in vivo, however, are not well defined. Since the expression of neurotrophin genes is considered an important component of the intrinsic neuroprotective response to insults, we analyzed the gene expression of neurotrophins in the brains of rats which received a continuous infusion of Abeta-(1-42) into the cerebroventricle. Northern blot analysis revealed a significant increase in brain-derived neurotrophic factor (BDNF) expression in the hippocampus but no change in the cerebral cortices. The alteration peaked on days 3-7 and returned to the basal level on day 14 after the start of Abeta-(1-42) infusion. No significant changes in nerve growth factor or neurotrophin-3 mRNA expression were observed. The infusion of Abeta-(1-40) and (25-35) also triggered the expression of BDNF mRNA, whereas neither Abeta-(40-1) nor (1-16) had any effect. In situ hybridization histochemistry revealed that the expression mainly occurred in the hilus and granular layer of the dentate gyrus and to a lesser extent in the pyramidal neurons of the CA1 region. These results demonstrate that the continuous intracerebroventricular infusion of Abeta induces selective and spatiotemporal expression of BDNF mRNA in the hippocampus. PMID- 11038252 TI - Activation of the Jak-Stat- and MAPK-pathways by oncostatin M is not sufficient to cause growth inhibition of human glioma cells. AB - We have recently described that oncostatin M (OSM), a member of the IL-6 family of cytokines, induces the differentiation of human glioma cells in culture. In order to extend this studies, we analyzed the effect of OSM on other human glioma cell lines including A172, U343-MG and T98G. All of these cell lines express the receptor components of OSM and leukemia inhibitory factor (LIF) gp130, LIFR and the OSM specific OSMRbeta. Therefore, we expected these cell lines to respond to OSM and LIF. Using specific antibodies recognizing proteins of the janus kinase (Jak-)/signal transducers and activator of transcription (Stat-) signaling cascade that has been shown to transduce the signals of the IL-6 cytokines to the nucleus, we could show that Jak1, Jak2 and Tyk2, as well as the Stat proteins Stat1, Stat3 and Stat5b were phosphorylated in all three cell lines by OSM and, at least in part, by LIF. Activation of the Stat proteins was also detected by EMSA which revealed complex formation on the Stat3 DNA-binding element and on a Stat5 binding site. Consistent with our recent findings, OSM treatment also induced the activation of the MAPK erk2 and the tyrosine phosphatase SHP-2 in cells of the A172, T98G and U343-MG cell lines. Although this activation pattern was very close to what we had observed in the GOS3 glioma cells, only T98G showed a growth inhibition in response to OSM while the A172 and the U343-MG cell lines did not respond to OSM treatment in terms of growth inhibition. PMID- 11038253 TI - The splicing determinants of a regulated exon in the axonal MAP tau reside within the exon and in its upstream intron. AB - Tau is a microtubule-associated protein whose transcript undergoes complex regulated splicing in the mammalian nervous system. Exon 6 of the gene is an alternatively spliced cassette whose expression profile is distinct from that of the other tau regulated exons, implying the utilization of distinct regulatory factors. Previous work had established the use of cryptic splice sites within exon 6 and the influence of flanking exons on the ratio of exon 6 variants. The present work shows that, in addition to the previously identified participants, the ratio of exon 6 isoforms is affected by: (1) suppression of the cryptic sites, (2) deletions of the upstream intron, and (3) the splicing regulators PTB and U2AF, both of which act on the branch point/polypyrimidine tract region. These results strongly suggest that factors binding immediately upstream of exon 6 are involved in regulation of this exon. They also lead to the conclusion that splicing of exon 6 is primarily governed by multiple branch points. PMID- 11038254 TI - Caspase-3 activation and inflammatory responses in rat hippocampus inoculated with a recombinant adenovirus expressing the Alzheimer amyloid precursor protein. AB - To elucidate the mechanism of neuronal death in Alzheimer's disease, we investigated the effects of overexpression of wild-type Alzheimer amyloid precursor protein (APP) on neuronal cells and glial cells in vivo. When an APP695 expressing adenovirus was injected into the dorsal hippocampal region, a number of neurons in remote areas were positively stained with anti-APP monoclonal antibody, and underwent severe degeneration from 3 to 7 days after viral inoculation. Most degenerating neurons were immunopositive with both APP and activated caspase-3, but some neurons that expressed activated caspase-3 were not expressing APP from 7 to 14 days after virus injection. In the neighborhood of the degenerating neurons, activated microglia/macrophages, which were identified by the phenotypic marker C3bi receptor (CD11b/c; OX-42), were observed, and some of them appeared to phagocytose the caspase-3-immunopositive degenerating neurons. In addition to microglia/macrophages, infiltrating leukocytes expressing CD45 or CD4 were also detected. These results suggest that the increased accumulation of APP induced not only caspase-3-mediated death machinery, but also inflammatory responses including microglial activation. These inflammatory responses might cause further neurodegeneration through the alternative pathway that might activate the caspase-3-mediated death machinery without APP expression. PMID- 11038255 TI - Expression of GABA(A) receptor alpha5 subunit-like immunoreactivity in human hippocampus. AB - To investigate the distribution of GABA(A) receptor alpha5 subunit expression in brain, polyclonal antisera were raised, characterised and applied to human and rat brain sections. The resultant antibodies detected a major band of 53 kDa in sodium dodecyl sulphate-polyacrylamide gel electrophoresis immunoblots. Abundant immunostaining was demonstrated in the hippocampal formation in multiple cell types, although predominantly in pyramidal neurons. These data are supportive of GABA-ergic involvement in cognition, and suggest that this influence may be mediated through receptors containing the alpha5 subunit. PMID- 11038256 TI - Aggregates of cAMP-dependent kinase RIalpha characterize a type of cholinergic neurons in the rat brain. AB - Acetylcholine is synthesized by different types of neurons, showing a distinct biochemical phenotype. Aggregates of RIalpha regulatory subunit of cAMP-dependent protein kinases are visualized by immunohistochemistry only in some cholinergic neurons, since they tightly colocalize with two different markers, choline acetyltransferase (ChAT) and vesicular acetylcholine transporter (VAChT). These neurons are present mainly in brain areas related to the limbic system. None of the other regulatory subunits of cAMP dependent kinases colocalize with cholinergic markers. PMID- 11038257 TI - Direct interactions of methamphetamine with the nucleus. AB - Possible direct effects of methamphetamine (METH) on transcription factors AP-1 and cAMP response element-binding protein (CREB) in the nucleus were assessed by electrophoretic mobility-shift assay. In vitro addition of METH to nuclear extract from brain tissue increased DNA-binding activities of both transcription factors. In addition, injections of METH to mice induced increases in the binding of AP-1 and CREB, which were depleted by preincubating the nuclear extract with anti-METH antibody. We also examined the cellular distribution of METH in mesencephalic neuronal cells using an immunofluorescence experiment with anti METH antibody. METH-like immunoreactivity was seen to accumulate in the cytosol 4 6 h after the METH treatment. Furthermore, METH-positive signals were also observed in the nuclei of the METH-treated cells. The present study is the first demonstration that METH can have direct effects on DNA-binding protein complex by redistributing not only in the cytosol but also into the nucleus. PMID- 11038258 TI - Differential RNA cleavage and polyadenylation of the glutamate transporter EAAT2 in the human brain. AB - We cloned four novel transcripts of the excitatory amino acid transporter 2, named EAAT2/3UT1-4, resulting from differential cleavage and polyadenylation. Tandem poly (A) sites were found to be functional at 72, 654, 973 nucleotides and more than 2 kb downstream of the stop codon. A tissue-specific expression was identified for 3'-variants of the EAAT2 RNA, most prominently for EAAT2/3UT4 (hippocampus>cortex>>cerebellum>thalamus) as demonstrated by Northern blot analysis and quantitative PCR. We conclude, that alternative poly (A) selection may contribute to the reported differential EAAT2 protein expression under normal and diseased conditions. PMID- 11038259 TI - Activity of adenosine deaminase in the sleep regulatory areas of the rat CNS. AB - There are data to support the notion that adenosine (ADO), a neuromodulator in the CNS, is an important regulator of sleep homeostasis. It has been demonstrated that ADO agonists and antagonists strongly impact upon sleep. In addition, the level of adenosine varies across the sleep/wake cycle and increases following sleep deprivation. Adenosine deaminase (ADA) is a key enzyme involved in the metabolism of ADO. We questioned, therefore, whether there are differences in adenosine deaminase activity in brain regions relevant to sleep regulation. We found that ADA exhibits a characteristic spatial pattern of activity in the rat CNS with the lowest activity in the parietal cortex and highest in the region of the tuberomammillary nucleus (15.0+/-4.8 and 63.4+/-28.0 nmoles/mg protein/15 min, mean+/-S.D., respectively). There were significant differences among the brain regions by one-way ANOVA (F=31.33, df=6, 123, P=0.0001). The regional differences in ADA activity correlate with variations in the level of its mRNA. This suggests that spatial differences in ADA activity are the result of changes in the expression of the ADA gene. We postulate that adenosine deaminase plays an important role in the mechanism that controls regional concentration of adenosine in the brain and thus, it is a part of the sleep-wake regulatory mechanism. PMID- 11038260 TI - Phylogenetic relation of lungfish indicated by the amino acid sequence of myelin DM20. AB - The cDNA of lungfish Protopterus annectens myelin DM20 was cloned, and the complete amino acid sequence of Protopterusannectens DM20 was deduced. When five possible phylogenetic trees were tested for the DM20 sequences, the maximum likelihood method supported tree 1 [((tetrapods, lungfish), coelacanth), zebrafish, shark] or tree 5 [(tetrapods, lungfish), (coelacanth, zebrafish), shark]. Both tree 1 and tree 5 indicate that lungfish is phylogenetically the closest to tetrapods among the living fishes. PMID- 11038261 TI - Differential expression of estrogen receptor beta splice variants in rat brain: identification and characterization of a novel variant missing exon 4. AB - Estrogen receptor beta (ER-beta) mRNA is found in abundance in rat brain. The distribution of ER-beta mRNA in brain differs from that of ER-alpha suggesting they subserve different functions. ER-beta mRNA has been reported to be variably spliced, in contrast to ER-alpha, resulting in numerous isoforms that possess different functional properties. The present study was undertaken to determine whether the isoforms of ER-beta mRNA are differentially distributed in different brain regions. In order to assess the range of transcript forms expressed in various brain regions in the same assay, a micropunch dissection technique was combined with semiquantitative RT-PCR. The relative abundance of each ER-beta isoform (beta1>beta2>beta1delta3>beta2delta3) was similar in all ER-beta positive brain regions with the exception of the hippocampus, which contained low levels of most isoforms and a fifth ER-beta isoform, which we are calling ER beta1delta4. Based on its sequence, ER-beta1delta4 encodes an ER-beta that is missing exon 4. Initial characterization studies of this showed that it did not bind estrogen, and that, unlike ER-beta1, it localized to the cytoplasm when expressed in cultured cells. The distribution of ER-beta1delta4 was different from that of the other isoforms in that it was expressed at high levels in the hippocampus, where the other isoforms were low, and that it was nearly undetectable in the brain regions that expressed the highest levels of the other ER-beta splice variants. These data suggest that a highly complex pattern of estrogen signaling can occur in a region specific manner in the rat brain. PMID- 11038262 TI - Cloning and functional expression of rKCNQ2 K(+) channel from rat brain. AB - By homologue cloning, we have isolated a cDNA encoding a voltage-gated K(+) channel, rKCNQ2, from a rat brain cDNA library using RACE. The open reading frame of the translated protein comprises 852 amino acids with 6 transmembrane segments and a pore motif between S5 and S6. rKCNQ2 shares 96% amino acid identity with human KCNQ2 in which mutations cause a form of epilepsy known as benign familial neonatal convulsions (BFNC). Northern blotting with a rKCNQ2-specific probe revealed a robust single band of 8.6-kb transcript expressed in brain not in other tissues. Functional expression of rKCNQ2 in an HEK 293 cell line by whole cell current recording and in Xenopus oocytes by two-electrode voltage clamp showed outward K(+) selective currents that displayed delayed rectifier-type kinetics. The G-V curve, fitted with a Boltzmann function, showed voltage dependence of activation with a threshold of activation approximately -60 mV. The rKCNQ2 currents were sensitive to TEA block with a Ki of 0.1 mM. In addition, rKCNQ2 currents were down-regulated upon exposure of cells to either a broad spectrum tyrosine kinase inhibitor genistein or a Src-like tyrosine kinase inhibitor herbimycin A. Our findings add a rodent member to the KCNQ channel subfamily, providing new information of the channel modulation, and will facilitate generation of rodent models of epilepsy. PMID- 11038263 TI - Post-transcriptional regulation of zenk expression associated with zebra finch vocal development. AB - In the male zebra finch, highly variable juvenile song and stereotyped adult song induce mRNA expression of the immediate early gene zenk in telencephalon. However, the functional consequences of this behavior-driven gene expression remain unknown. Here we characterize the developmental expression of zenk mRNA and protein in two forebrain song regions (HVC, the higher vocal center, and RA, the robust nucleus of the archistriatum). In HVC, singing results in similar percentages of cells producing zenk mRNA and zenk protein at different stages of vocal development. Similarly, song behavior at all stages of vocal development induces a comparable percentage of RA cells expressing zenk mRNA. However, the percentage of RA zenk immunoreactive cells is low during early vocal learning, increasing only as the vocal pattern matures. Early induction of a stereotyped vocal pattern in juvenile birds is associated with increased zenk immunoreactivity in RA, indicating that it is the form of the behavior (and not the age of the bird) that correlates with changes in zenk immunoreactivity. Together, our findings reveal a previously unrecognized relationship between behavioral development and post-transcriptional gene regulation. PMID- 11038264 TI - Human cytomegalovirus IE1 promoter/enhancer drives variable gene expression in all fiber types in transgenic mouse skeletal muscle. AB - BACKGROUND: Versatile transgenic manipulation of skeletal muscle requires knowledge of the expression profiles of diverse promoter/enhancer elements in the transcriptionally specialized fiber types of which muscle is composed. "Universal" viral promoters/enhancers, e.g., cytomegalovirus IE1 (CMV IE1), are of interest as reagents that may drive broad expression. However, a previous study noted a marked heterogeneity of CMV IE1-driven transgene expression among muscle fibers, raising the possibility of fiber-type-restricted expression. The purpose of the present study was to characterize CMV IE1-driven expression in terms of fiber type. RESULTS: We produced two lines of transgenic mice carrying the CMV IE1/ beta-galactosidase construct CMVLacZ, and analyzed transgene expression and fiber type by histochemical analysis of hindlimb muscle sections. In both lines CMVLacZ was expressed in all four major fiber types: type I (slow) and types IIA, IIB and IIX (fast). There was no unique pattern of fiber-type preferential expression; fiber-type quantitative differences were observed but details varied between muscle regions and between lines. Both lines showed similar fiber-type-independent regional differences in overall expression levels, and a high level of within-fiber-type variability of expression, even among nearby fibers. The soleus muscle showed strong expression and comparatively little within-fiber-type or between-fiber-type variability. CONCLUSIONS: The CMV IE1 promoter/enhancer is not fiber-type-restricted and can be useful for driving germ-line transgene expression in all four fiber types. However, not all fibers express the gene at high levels due in part to regional differences in overall expression levels, and to a high level of within-fiber-type variability. Given the multinucleate syncitial nature of muscle fibers, it is not likely that this variability is due to variegating heterochromatinization. The soleus muscle would make a suitable subject for near-uniform experimental gene expression driven by CMV IE1 elements. PMID- 11038265 TI - Are the pneumococcal polysaccharide vaccines effective? Meta-analysis of the prospective trials. AB - The objective was to review the evidence of effectiveness of the polyvalent polysaccharide pneumococcal vaccine from prospective properly randomised controlled trials comparing pneumococcal vaccines with placebo in subjects who are immunocompetent and those likely to have an impaired immune system. Databases searched included the Cochrane Library, (issue 2, 2000), MEDLINE (1966-August 2000), PubMed (to August 2000) and EMBASE ( to August 2000). Reference lists of reports and reviews were also searched. To be included in the analysis, a study had to have been a prospective randomised comparison of a polysaccharide pneumococcal vaccine (any valency) and to have a placebo or no treatment comparison group. Papers had to report important clinical outcomes, such as rates of pneumonia, pneumococcal pneumonia, lower respiratory tract infections, pneumonia deaths or bacteraemia. Serological outcomes were not sought. Thirteen randomised comparisons with over 45,000 subjects were identified in an extensive literature review. Eight studies had a quality score of 3 or more on a scale of 1 to 5. In three comparisons with 21,152 immunocompetent subjects (South African gold miners, New Guinea highlanders) pneumococcal vaccination was effective in reducing the incidence of all-cause pneumonia (relative risk 0.56, 95% confidence interval 0.47 to 0.66), pneumococcal pneumonia (0.16; 0.11 to 0.23), pneumonia deaths (0.70; 0.50 to 0.96) and bacteraemia (0.18; 0.09 to 0.34). In ten comparisons in over 24,000 people who were elderly or likely to have impaired immune systems, pneumococcal vaccination was without effect for any outcome. Present guidelines recommend pneumococcal vaccination for "high-risk" groups. There is no evidence from randomised trials that this is of any benefit. PMID- 11038266 TI - Early peri-operative hyperglycaemia and renal allograft rejection in patients without diabetes. AB - BACKGROUND: Patients with diabetes have an increased risk for allograft rejection, possibly related to peri-operative hyperglycaemia. Hyperglycaemia is also common following transplantation in patients without diabetes. We hypothesise that exposure of allograft tissue to hyperglycaemia could influence the risk for rejection in any patient with high sugars. To investigate the relationship of peri-operative glucose control to acute rejection in renal transplant patients without diabetes, all patients receiving their first cadaveric graft in a single center were surveyed and patients without diabetes receiving cyclosporin-based immunosuppression were reviewed (n = 230). Records of the plasma blood glucose concentration following surgery and transplant variables pertaining to allograft rejection were obtained. All variables suggestive of association were entered into multivariate logistic regression analysis, their significance analysed and modeled. RESULTS: Hyperglycaemia (>8.0 mmol/L) occurs in over 73% of non-diabetic patients following surgery. Glycaemic control immediately following renal transplantation independently predicted acute rejection (Odds ratio=1.08). 42% of patients with a glucose < 8.0 mmol/L following surgery developed rejection compared to 71% of patients who had a serum glucose above this level. Hyperglycaemia was not associated with any delay of graft function. CONCLUSION: Hyperglycaemia is associated with an increased risk for allograft rejection. This is consistent with similar findings in patients with diabetes. We hypothesise a causal link concordant with epidemiological and in vitro evidence and propose further clinical research. PMID- 11038267 TI - Prediction of tight turns and their types in proteins. AB - A tight turn in protein structure is defined as a site where (i) a polypeptide chain reverses its overall direction, i.e., leads the chain to fold back on itself by nearly 180 degrees, and (ii) the amino acid residues directly involved in forming the turn are no more than six. Tight turns are generally categorized as delta-turn, gamma-turn, beta-turn, alpha-turn, and pi-turn, which are formed by two-, three-, four-, five-, and six-amino-acid residues, respectively. According to the folding mode, each of such tight turns can be further classified into several different types. Tight turns play an important role in globular proteins from both the structural and functional points of view. In view of this, various efforts have been made to predict tight turns and their types. This Review summarizes the development in this area, with an emphasis focused on the most recent work concerned that is featured by the sequence-coupled model. Meanwhile, the future challenge in this area has also been briefly addressed. PMID- 11038268 TI - Europium cryptate-tethered ribonucleotide for the labeling of RNA and its detection by time-resolved amplification of cryptate emission. AB - TRACE (time-resolved amplification of cryptate emission), also called HTRF for pharmaceutical applications, is a homogeneous time-resolved fluorescence technique well adapted for the study of molecular interactions. It is based on fluorescence resonance energy transfer (FRET) between europium trisbipyridine cryptate (TBPEu(3+)) as energy donor and cross-linked allophycocyanin, symbolized by XL665, as acceptor, leading to a long-lived FRET signal. TBPEu(3+)-labeled uridine triphosphate (UTP), referred to as K-11-UTP in the text, was obtained by coupling TBPEu(3+) moiety to a C-5 functionalized UTP analog. K-11-UTP can be directly incorporated in RNA strands during enzymatic synthesis. This was demonstrated in an in vitro transcription reaction promoted by T(7) RNA polymerase. The reaction was performed in the presence of K-11-UTP and biotin labeled cytidine triphosphate (biotin-16-CTP) in admixture with natural ribonucleotides. After the addition of streptavidin-XL665 conjugate (SA-XL665), which binds on biotinylated cytidine residues, a long-lived FRET signal was obtained. This proved that both europium cryptate and biotin were incorporated into the same RNA strand and are close enough to generate a FRET signal. The study of this FRET detection assay format showed that such doubly labeled RNA can be easily detected even when a very low percentage of K-11-UTP is used (less than 1% of total UTP concentration). Europium-cryptate-labeled RNA can also be monitored using a homogeneous hybridization assay format involving a biotinylated probe. After the addition of SA-XL665, the FRET signal generated demonstrates the formation of RNA:DNA hybrids. Europium-cryptate-labeled nucleotide thus gives access to a new type of RNA nonisotopic labeling and homogeneous detection assays. PMID- 11038270 TI - Monochlorobimane fluorometric method to measure tissue glutathione. AB - Glutathione (GSH) is the principal intracellular low-molecular-weight thiol and plays a critical role in the cellular defense against agents that impose oxidative stress. A common technique to measure GSH uses reversed-phase high performance liquid chromatography (HPLC) following derivatization with 5, 5' dithiobis(2-nitrobenzoic acid), a technique, although reliable and sensitive, that is time consuming and laborious. A common technique to measure GSH in cultured cells is to add monochlorobimane to the culture medium where it readily enters cells to form a fluorescent GSH-monochlorobimane adduct that can be measured fluorometrically. This reaction is catalyzed by glutathione S transferase. We reasoned that adding glutathione S-transferase and monochlorobimane to tissue homogenates would allow a rapid reliable method to measure GSH. The accuracy of the new test was assessed in homogenates of rat livers. One-half of each homogenate was assayed for GSH using a HPLC approach while the other half was assayed using the monochlorobimane approach. The two methods were found to give identical results. We conclude that the monochlorobimane fluorescent method is sufficiently specific to reliably measure tissue GSH. PMID- 11038269 TI - Design of helical proteins for real-time endoprotease assays. AB - Proteases play a key role in cellular biology and have become priority targets for new pharmaceuticals. Thus, there is a high demand for specific, sensitive, and quick assays to monitor the activity of endoproteases. We designed affinity tagged helical proteins with unique protease cleavage sites and thus constructed universal, molecularly defined, and uniform substrates for in vitro detection of IgA endoprotease. The substrate is a 10.5-kDa recombinant helical protein with a high-affinity (His)(6)-tag at the amino-terminal end. Further elements are a unique proteolytic recognition site and a C-terminal helical extension, which is cut off by the protease. Proteolytic action can be monitored in real time using surface plasmon resonance spectroscopy. Femtomole amounts of protease could be reliably and quantitatively detected within a few minutes after the start of the reaction. The detection signal changed linearly with the amount of protease and was independent of the applied sample flow rate. The biochip can be reversibly loaded with the recombinant protease substrate, so that the SPR assay is well suited for automation. By substituting an HIV protease site for the recognition site of the IgAse, we also obtained a substrate for the quantitative and sensitive detection of HIV-1 endoprotease. Our substrate design is thus generally applicable. PMID- 11038271 TI - Enzymatic synthesis and purification of caffeoyl-CoA, p-coumaroyl-CoA, and feruloyl-CoA. AB - An enzyme preparation from wheat seedlings containing p-coumaroyl:CoA ligase activity was used to synthesize caffeoyl-CoA, p-coumaroyl-CoA, and feruloyl-CoA. The same enzyme preparation also contains caffeic acid-3-O-methyl transferase and caffeoyl-CoA-3-O-methyl transferase activities. The maximum activity was found in enzyme preparation from 2-day-old seedlings, where 15-20% of the hydroxy cinnamic acid could be converted into the corresponding thioester. This yield is a result of an equilibrium between the ligase and a thioesterase also present in the crude enzyme preparation. The activity of caffeic acid 3-O-methyl transferase and caffeoyl-CoA 3-O-methyl transferase enables the production of (14)C-labeled feruloyl-CoA when using S-adenosyl-l-[methyl-(14)C]-methionine as methyl donor. The produced thioesters can be purified by reverse phase HPLC using a phosphoric acid-acetonitrile gradient. PMID- 11038272 TI - High-throughput screening of enzyme inhibitors: simultaneous determination of tight-binding inhibition constants and enzyme concentration. AB - Active site titration by a reversible tight-binding inhibitor normally depends on prior knowledge of the inhibition constant. Conversely, the determination of tight-binding inhibition constants normally requires prior knowledge of the active enzyme concentration. Often, neither of these quantities is known with sufficient accuracy. This paper describes experimental conditions under which both the enzyme active site concentration and the tight-binding inhibition constant can be determined simultaneously from a single dose-response curve. Representative experimental data are shown for the inhibition of human kallikrein. PMID- 11038273 TI - Binding and detection of glycosaminoglycans immobilized on membranes treated with cationic detergents. AB - Immobilization of molecules on surfaces is used for preparative, quantitative, and qualitative studies. Glycosaminoglycans (GAGs) are strongly hydrophilic and negatively charged molecules that do not bind well to either polystyrene surfaces or hydrophobic blotting membranes. Hydrophobic membranes were derivatized with cationic detergents to become hydrophilic and positively charged. The ability of the polyvinylidene fluoride and nitrocellulose membranes to retain GAGs increased up to 12.8 microg per spot in the dot blot assay when the membrane was treated with a cationic detergent. Immobilized GAGs were stained with alcian blue, and the staining intensity was quantitated by scanning and densitometry. The derivatized membranes were used for solid-phase extraction of GAGs in blood plasma, urine, or cerebrospinal fluid. The detection sensitivity was equal for different types of GAGs but there was a slight negative interference from fibrinogen in blood plasma. The immobilized GAGs could also be released from the membrane using a nonionic detergent at high ionic strength. Recovery of different proteoglycan populations, separated by electrophoresis and detected by reversible staining with toluidine blue, was 70-100%. PMID- 11038274 TI - Detection of triplet repeat expansion in the human genome by use of hybridization signal intensity. AB - Triplet repeat disease is a group of hereditary neurodegenerative disorders caused by expansion of trinucleotide repeats such as CAG/CTG, CGG/CCG, and GAA/TTC. Direct detection of the expansion in the patient's genome shortcuts the tedious process needed for identification of disease genes by conventional approaches. Here we describe a method to detect triplet repeat expansion from the hybridization signal intensity. Using a digoxigenin-labeled (CTG)9 probe, the hybridization intensity and number of repeats showed a good linear correlation. The technique detected expansion in genomic DNA in all cases with moderate or large expansion. Even in the case of a small expansion, this method could detect the mutant fragment. The technique has advantages over related techniques because it is more sensitive and can be applied to cases where a small repeat expansion is involved. PMID- 11038275 TI - Near-infrared spectrophotometric determination of tri- and tetrapeptides. AB - A new method based on the near infrared technique has been developed for the noninvasive and nondestructive determination of the identity and sequences of amino acid residues in small peptides. The method is capable of distinguishing not only peptides with very similar structures (e.g., Gly-Ala-Ala, Gly-Ala-Leu, Leu-Gly-Gly and Gly-Leu-Leu-Gly, Gly-Leu-Gly-Gly, Gly-Gly-Ala-Gly) but also peptides with the same amino acid residues but different sequences (e.g., Gly-Ala Ala, Ala-Gly-Ala, Ala-Ala-Gly and Gly-Gly-Gly-Ala, Gly-Gly-Ala-Gly). PMID- 11038276 TI - A spectrophotometric method for determining the oxidative deamination of methylamine by the amine oxidases. AB - Previously published studies on the oxidative deamination of methylamine by the amine oxidases have determined the formation of radioactively labeled formaldehyde from [(14)C]methylamine. The present work describes a coupled spectrophotometric assay, using formaldehyde dehydrogenase, for the continuous determination of the oxidative deamination of methylamine by semicarbazide sensitive amine oxidase (SSAO) and its potential use for determining methylamine concentrations in plasma. In this assay, the formaldehyde produced by methylamine deamination is further oxidized to formate, with the reduction of NAD(+), by formaldehyde dehydrogenase. The NADH generated is monitored continuously at 340 nm. Interference from the presence of a rotenone-insensitive NADH oxidase activity in crude tissue homogenates and microsomal fractions can be minimized by pretreating samples with Triton X-100 or substituting NAD(+) by APAD(+) in the coupled assay. This relatively inexpensive and reproducible assay procedure avoids the use of radioactively labeled material. PMID- 11038277 TI - Effective isolation of high-quality total RNA from human adult articular cartilage. AB - The isolation of large quantities of good-quality RNA from human articular cartilage has been a long-standing problem for researchers working with human articular cartilage. In this paper we report a protocol which we have developed based on the Qiagen RNeasy procedure to produce high yields of purified, DNA-free RNA from normal and osteosteoarthritic human articular cartilage. The average yield of RNA was 8.39 microg/g (n = 59) for normal and 6.69 microg/g (n = 58) for osteoarthritic cartilage (average ratio OD 260/280 = 1.8-1.9). Quantitative PCR, cDNA array technology, and Northern blot analysis were used to verify the quality of the RNA. PMID- 11038278 TI - Assay of diverse protease activities on the basis of a small synthetic substrate. AB - The available methods for assaying of protease activity of unknown or partially defined specificity utilize long peptides, mostly denatured proteins, comprehending at least all coded amino acids in cleavable positions. In contrast, here we report on an alternative approach which utilizes a small synthetic ester substrate containing only one amino acid. The approach equally detects endo- and carboxypeptidases with a wide variety of specificities including enzymes specific for basic, acidic, aromatic, and nonaromatic hydrophobic amino acid moieties. The results further revealed that most proteases could be detected in activities considerably less than 1 U. In contrast to the methods used thus far, the enzymatic hydrolysis of the small substrate can be easily and rapidly assayed by a shift in absorption resulting in a change in color of the assay mixture at visible wavelengths. Thus, no additional instrumental efforts are generally required. On the basis of these characteristics, the approach presented here could be particularly valuable for monitoring the purification of enzymes or as a rapid check for protease activity. PMID- 11038279 TI - Identification of differentially expressed genes by mutually subtracted RNA fingerprinting. AB - A mutually subtracted RNA fingerprinting (SuRF) method has been developed that allows efficient identification of differentially expressed sequence tags between two samples. Mutual subtractions of two RNA samples are achieved by first synthesizing cDNAs using oligo(dT) coupled with magnetic beads which are then reciprocally hybridized to starting RNA samples to remove common mRNAs between them. The second step involves differential fingerprinting of the subtracted RNA samples by polymerase chain reaction with specially designed degenerate primers. SuRF was applied to identify alteration in gene expression pertinent to osteogenic sarcoma which was achieved by employing the method between FOB (an immortalized fetal osteoblast) and MG63 (an osteosarcoma) cell lines. An estimated 10% of the total expressed genes in these two cell types were screened by the method. This analysis identified 96 differentially expressed sequences, none of which was identified repeatedly. A subset of these sequences was subsequently confirmed to have differential expression between the two cell types. Removal of common mRNAs prior to differential display should diminish redundant identification of abundant genes and increase the chance of identifying rare differentially expressed genes. PMID- 11038280 TI - Ultramicroanalysis of reducing carbohydrates by capillary electrophoresis with laser-induced fluorescence detection as 7-nitro-2,1,3-benzoxadiazole-tagged N methylglycamine derivatives. AB - A method for ultramicroanalysis of carbohydrates using capillary electrophoresis with laser-induced fluorescence detection was developed, based on precapillary conversion to 7-nitro-2,1, 3-benzoxadiazole (NBD)-tagged N-methylglycamines. Although the derivatization involves two-step reactions, i.e., reductive N methylamination followed by condensation with NBD-F, they can be carried out in a one-pot fashion and proceed quantitatively within ca. 50 min in total. Since the reaction conditions are mild, it does not cause desialylation. The derivatives can be well separated by capillary electrophoresis and sensitively detected by argon laser-induced fluorescence. It allowed detection of monosaccharides of down to nanomolar concentrations for analytical sample solution, which correspond to the attomole injected amounts, and good linearity was observed over a wide range. It was also successfully applied to analysis of N-glycans in a microgram quantity of a glycoprotein. Studies on the cleanup of derivatized product are also described in relation to N-glycan analysis. PMID- 11038281 TI - Development of an internally quenched fluorescent substrate selective for endothelin-converting enzyme-1. AB - Endothelin-converting enzyme-1 (ECE-1) is a membrane-bound zinc-metallopeptidase that is related to neprilysin in amino acid sequence. A major in vivo function of ECE-1 is the proteolytic conversion of big endothelin-1 to endothelin-1, one of the most potent vasconstricting peptides known. Although ECE-1 was once thought to be specific for the processing of endothelin precursors, it is now known that the enzyme hydrolyzes a number of peptide hormones. We have incorporated knowledge gained from recent studies of ECE-1 substrate specificity to aid the design of internally-quenched fluorescent substrates derived from bradykinin. The best of these substrates, (7-methoxycoumarin-4-yl)acetyl-Arg-Pro-Pro-Gly-Phe-Ser Ala-Phe-Lys(2, 4-dinitrophenyl), is hydrolyzed by ECE-1 with a k(cat)/K(m) value of 1.9 x 10(7) M(-1) s(-1), making it the most sensitive substrate yet described for ECE-1. The substrate is suitable for the rapid, continuous assay of the enzyme using a microplate format in a fluorescence plate reader, thereby simplifying both the purification of ECE-1 and the characterization of its inhibitors. It is demonstrated that (7-methoxycoumarin-4-yl)acetyl-Arg-Pro-Pro Gly-Phe-Ser-Ala-Phe-Lys(2, 4-dinitrophenyl) is also a substrate for neprilysin, but is hydrolyzed 10-fold more efficiently by ECE-1, making this substrate selective for ECE-1. Furthermore, this synthetic peptide is a poor substrate for the matrix metalloproteinases. PMID- 11038282 TI - Applying phage antibodies to proteomics: selecting single chain Fv antibodies to antigens blotted on nitrocellulose. AB - Two-dimensional gel electrophoresis is a powerful tool for identification of proteins that differ between patients with qualitatively or quantitatively different disease states. Further characterization of these protein differences would be greatly facilitated by the availability of antibodies that could be used to detect and quantitate the temporo-spatial pattern and cellular and tissue location of the different proteins. To generate such antibodies, methods were developed which permit the successful selection of monoclonal phage antibodies from phage display libraries against antigens blotted from SDS-PAGE gels onto nitrocellulose. First, it was determined that nitrocellulose and PVDF membranes gave significantly lower levels of background phage binding than two other membranes studied. Next, it was determined that blocking with fish gelatin and binding in the presence of 0.5 M NaCl could reduce nonspecific binding 10,000 fold and result in enrichment ratios greater than 500-fold with antigen concentrations as low as 1 ng/mm(2). When optimized conditions were applied to phage antibody libraries, panels of monoclonal phage antibodies were generated against the proteins ErbB2 and bovine serum albumin electroblotted from SDS-PAGE gels onto nitrocellulose. Antibodies were obtained with as little as 10 to 1 ng of antigen, depending on whether the libraries displayed single or multiple copies of antibody per phage. The antibodies worked as reagents in both ELISA and Western blotting. PMID- 11038283 TI - Human ras-converting enzyme (hRCE1) endoproteolytic activity on K-ras-derived peptides. AB - A human gene responsible for one of the steps in Ras post-translational modification and membrane localization, hRCE1, encodes a 35-kDa membrane associated endoprotease. We examined hRCE1 activity using farnesylated 9 aa peptides with the core sequence, KSKTKC(farnesyl)VIM [(farnesyl) = (f)], from the C-terminus of K-Ras. We first demonstrated hRCE1 specificity in cleavage location and endoproteolysis. We then describe a direct fluorescent microtiter plate assay. We demonstrated that hRCE1 protease cleaved KSKTKC(f)VIM peptides between the C(f) and V positions, generating KSKTKC(f) and the corresponding tripeptides as products. We found that the sequence KSKTKC(f)VI was a better substrate for hRCE1 than KSKTKC(f)VIM. We also found that hRCE1 cleaved modified versions of KSKTKC(f)VIM that incorporated either MCA or ABZ fluorescent chromophores at the N-terminus, and quenching-group-containing amino acids at the V or M, but not the I, amino acid positions of VIM. The quenching-group-containing amino acids used were either Q(S) (dinitrophenyldiaminopropionic acid) or Q(L) (lysine epsilon dinitrophenyl). Cleavage of KSKTKC(f)VIM and modified versions of this peptide by hRCE1 was initially evaluated by HPLC product resolution and quantitation. The hRCE1 cleavage of quenched peptides enabled us to directly monitor proteolytic activity in a 96-well microtiter fluorescent plate assay. The microtiter format assay was validated by its sensitivity to RPI, an inhibitor of prenyl protein protease. A direct fluorescent assay provides an effective tool for further characterization of this enzyme and also for detection of novel inhibitors. PMID- 11038284 TI - Development of a mechanism-based, DNA staining protocol using SYTOX orange nucleic acid stain and DNA fragment sizing flow cytometry. AB - Accurate measurement of single DNA fragments by DNA fragment sizing flow cytometry (FSFC) depends upon precise, stoichiometric DNA staining by the intercalating dye molecules. In this study, we determined the binding characteristics of a commercially available 532 nm wavelength-excitable dye and used this information to develop a universal DNA staining protocol for DNA FSFC using a compact frequency-doubled Nd:YAG laser excitation source. Among twelve 532 nm wavelength-excitable nucleic acid staining dyes tested, SYTOX Orange stain showed the highest fluorescence intensity along with a large fluorescence enhancement upon binding to double-stranded DNA ( approximately 450-fold). Furthermore, using SYTOX Orange stain, accurate fragment-size-distribution histograms were consistently obtained without regard to the staining dye to base pair (dye/bp) ratio. A model describing two binding modes, intercalation (primary, yielding fluorescence) and external binding (secondary, involving fluorescence quenching), was proposed to interpret the performance of the dyes under different dye/bp ratios. The secondary equilibrium dissociation constant was found to be the most critical parameter in determining the sensitivity of each fluorophore to the staining dye/bp ratio. The measurements of both equilibrium dissociation constants provided us with a theoretical framework for developing a universal protocol which was successfully demonstrated over a wide range of DNA concentrations on a compact flow cytometer equipped with a frequency doubled, diode-pumped, solid-state Nd:YAG laser for rapid and sensitive DNA fragment sizing. PMID- 11038285 TI - Determination of matrix metalloproteinase activity using biotinylated gelatin. AB - Matrix metalloproteinases (MMPs) and, specifically, MMP-2 (gelatinase A) and MMP 9 (gelatinase B) are strongly associated with malignant progression and matrix remodeling. These enzymes are a subject of intensive studies involving screening of comprehensive chemical libraries of synthetic inhibitors. There is no simple method available for measurement of activity of gelatinases and related MMPs. Here, we report a simple, inexpensive, and highly sensitive assay for MMP activity. The assay performed in a 96-well microtiter plate format employs biotin labeled gelatin (denatured collagen type I) as a substrate. Following the substrate cleavage, only the proteolytic fragments bearing biotin moieties are captured by streptavidin coated on the plastic surface and the captured fragments with at least two biotin molecules should be revealed by streptavidin conjugated with horseradish peroxidase. The frequency of lysine residues is low in collagen type I relative to the MMP cleavage sequences (PXGX). Accordingly, the majority of the cleavage products must be devoid of biotin or possess only one biotin group. Both of these types of fragments cannot be recognized by the horseradish peroxidase-streptavidin conjugate. Therefore, higher gelatinolytic activity is associated with lower signal in the assay. This 2-h assay allows identification of gelatinolytic activity of MMP-2 in concentrations as low as 0.16 ng/ml. The sensitivity of this ELISA-like assay is comparable to that of gelatin zymography, a method widely used to detect gelatinases. However, in contrast to zymography, the assay directly measures the enzymatic activity of MMP samples. The gelatinolytic activity assay permits efficient analyses and screening of the MMP inhibitor panels and allows quantitation of gelatinolytic activity of various MMPs in solution as well as on cell surfaces. PMID- 11038286 TI - Rapid spectrophotometric determination of oxygen consumption using hemoglobin, in vitro: light scatter correction and expanded dynamic range. AB - The method of using absorbance in conjunction with hemoglobin (Hb) to monitor rapid changes in oxygen consumption in vitro was improved by using a non-linear calibration technique and multiwavelength spectroscopy. The O(2) dependence of Hb absorbance was effectively linearized using the current technique (R(2) = 0.990+/ 0.002, n = 3), and extended the dynamic range of [O(2)] determinations by 1.6 fold over previous approaches. The association/dissociation rates of O(2) and Hb were evaluated using the current approach and were not significant on the 100-ms time domain. A method was also developed for compensating for large amplitude light scattering changes in turbid media using multiwavelength analysis. Both the nonlinear calibration curve and light scattering corrections were validated in isolated porcine heart mitochondrial preparations. PMID- 11038287 TI - Luminometric quantitation of photinus pyralis firefly luciferase and Escherichia coli beta-galactosidase in blood-contaminated organ lysates. AB - Firefly luciferase and Escherichia coli beta-galactosidase chemiluminescent reporter gene assays are rapid and sensitive means of detecting reporter enzyme activities in cell lysates of both eukaryotic and prokaryotic systems. In these assays, expression vectors containing the luciferase or beta-galactosidase genes are transferred to cells in culture or animal tissues in vivo. Crude cell or organ lysates are then prepared and submitted to enzyme assays. The level of enzyme activity is proportional to the efficiency of gene delivery and expression. When used with modified substrates that emit light when cleaved by the appropriate enzyme, luciferase and beta-galactosidase activity can be detected luminometrically. Attempts to apply these assays to cell lysates contaminated with blood, as from any whole organ lysate, have had questionable results thus far because of light absorption by hemoglobin in the ranges of light emission by both of these assays. We have made several adjustments to standard chemiluminescent reporter gene assay protocols to minimize errors in quantitation contributed by hemoglobin. To this end, we have developed a method for quantitating the protein due to blood and due to the organ itself in a blood contaminated organ lysate. We have also found that the use of a colorimetric protein assay that is unaffected by hemoglobin absorbance is preferred for protein quantitation. In conclusion, luciferase and beta-galactosidase assays can be applied to blood-contaminated organ lysates; however, the luciferase assay proved to be superior due to minimal endogenous activity and lower absorption by hemoglobin of light emitted by the enzyme product. PMID- 11038288 TI - Two time-resolved fluorometric high-throughput assays for quantitation of GDP-L fucose. AB - Two rapid and simple procedures for the quantitative analysis of GDP-l-fucose (GDP-Fuc) are described. The methods are based on time-resolved fluorescence and microplate assay technology. The first assay relies on measuring the enzyme activity of alpha1, 3-fucosyltransferase. In this assay, transfer of fucose from GDP-Fuc converts sialyllactosamine to sialyl Lewis x tetrasaccharide, which is detected and quantified by relevant antibodies on a microplate. The formation of the reaction product is directly dependent on the presence of GDP-Fuc in the concentration range of 10-10,000 nM. In the second method GDP-Fuc inhibits the binding of fucose-specific Aleuria aurantia lectin to fucosylated glycan on a microwell. The lectin-based assay is less sensitive than the enzyme assay, but it is cheaper and faster. We used these assays in monitoring the amount of GDP-Fuc in crude lysates of transgenic yeast, which expresses the enzymes producing GDP Fuc. The newly developed assays are versatile and applicable to measure also other nucleotide sugars or glycosyltransferase activities in a high-throughput manner. PMID- 11038289 TI - Prolactin and parental behavior in Adelie penguins: effects of absence from nest, incubation length, and nest failure. AB - Adelie penguin (Pygoscelis adeliae) males and females, nesting in Antarctica, alternate attendance at the nest with absences of many days to forage at sea. We investigated the importance of tactile input from egg and chicks on prolactin levels by observing nest attendance patterns and obtaining blood samples (1) during the first nest exchange of the incubation stage, (2) from birds whose incubation period was artificially increased or decreased by about 10 days, and (3) from birds whose nests had failed. Prolactin levels in females after 8 to 11 days of absence from the breeding colony did not differ from those in incubating males and did not change after females resumed incubation. Moving eggs between nests resulted in nests in which chicks hatched after about 26, 36 (normal), or 46 days. Duration of incubation did not affect prolactin levels in the parents measured during incubation, at the pip stage, hatch stage, or early brood stage. Adults first left their chicks unguarded on about the same calendar date, regardless of chick age. However, chicks from long incubation nests averaged 8 days younger when they were left unguarded than chicks from control or short incubation nests. In females, there was no effect of nest failure on prolactin levels. In males, prolactin levels were slightly lower after nest failure than in males tending nests. Testosterone was significantly higher in males after nest failure than in males still tending nests. Prolactin is elevated in Adelie penguins as part of the program of cyclical hormonal changes that accompany the lengthy reproductive season and is relatively independent of tactile input. Sustained prolactin secretion is probably required for the maintenance of parental behavior in offshore feeding species that must be absent from the nest for many days at a time. PMID- 11038290 TI - Preparental hormone levels and parenting experience in male cotton-top tamarins, Saguinus oedipus. AB - Male cotton-top tamarins, Saguinus oedipus, display hormonal changes associated with parenting prior to the birth of their infants. Here we examined the hormonal changes that occurred in experienced and inexperienced fathers during the postconception period, prior to the birth of infants. Noninvasive techniques were used to collect urine from 10 male cotton-top tamarins (5 experienced and 5 inexperienced breeders) three times weekly during the 6-month gestation period. Samples were analyzed for prolactin, testosterone, dihydrotestosterone (DHT), and cortisol, averaged by gestational month. Experienced males showed elevated prolactin during the mate's 3rd gestational month, and the elevation correlated with the number of infants surviving from the previous birth (0, 1, and 2) but not with outcome of the current pregnancy. However, an experienced male with no infants present still showed elevated prolactin and some inexperienced males showed elevated prolactin just before parturition, suggesting noninfant cues are also important. While prolactin levels were influenced by the male's prior infant experience, testosterone levels did not differ between experienced and inexperienced males. Testosterone levels were significantly elevated for all males during the 3rd, 4th and 5th months but had no relationship with number of infants present or with outcome of current pregnancy. DHT decreased during the second half of pregnancy compared with testosterone but this finding was not consistent for every male. No significant changes occurred in cortisol levels. These results suggest that infant-rearing experience affected the hormonal responsitivty of the male to his mate's current pregnancy. PMID- 11038291 TI - The role of prolactin in the regulation of clutch size and onset of incubation behavior in the American kestrel. AB - In most bird species, the timing of incubation onset may influence the degree of hatching asynchrony, which, together with variation in clutch size, affects reproductive success. In some domesticated species that usually show no hatching asynchrony, plasma prolactin concentrations in females rise with the onset of incubation and the end of laying, and this rise enhances incubation behavior and may terminate laying. To investigate whether a rise in prolactin during laying is involved in the regulation of clutch size and incubation onset in a species with hatching asynchrony, we measured plasma concentrations of immunoreactive prolactin (ir-prolactin) in laying American kestrels, Falco sparverius, and quantified clutch size and incubation behavior. In a separate study, we administered one of three concentrations of ovine prolactin (o-prolactin) via osmotic pumps implanted in females when egg 2 of a clutch was laid. ir-Prolactin concentrations during laying were higher in small than in large clutches and increased in parallel with the development of incubation behavior. o-Prolactin treatment enhanced incubation behavior, but did not affect clutch size, possibly because the manipulation was performed after clutch size had already been determined. Consistent with studies on domesticated species that show synchronous hatching, our results indicate that rising prolactin during laying enhances the expression of incubation behavior in a species that shows hatching asynchrony. Further studies are necessary to determine whether the relationship between prolactin and clutch size in the American kestrel is one of causation or of mere association. PMID- 11038292 TI - Androgen correlates of socially induced changes in the electric organ discharge waveform of a mormyrid fish. AB - Weakly electric fish from the family Mormyridae produce pulsatile electric organ discharges (EODs) for use in communication. For many species, male EODs are seasonally longer in duration than those of females, and among males, there are also individual differences in EOD duration. While EOD elongation can be induced by the administration of exogenous androgens, androgen levels have never before been assessed under natural or seminatural conditions. By simulating the conditions occurring during the breeding season in the laboratory, we provide evidence of a sex difference in EOD duration as well as document levels of circulating androgens in males. In this study, we analyzed the nature of social influences on male EOD duration and plasma androgen levels in Brienomyrus brachyistius. Individual males, first housed with a single female and then placed into social groups consisting of three males and three females, showed status dependent changes in EOD duration. Top-ranking males experienced a relatively large increase in EOD duration. Second-ranking males experienced a more modest increase, and low-ranking males experienced a decrease in EOD duration. These changes were paralleled by differences in circulating levels of plasma 11 ketotestosterone (11-KT), but not testosterone, suggesting that the changes in EOD duration may have been mediated by changes in plasma 11-KT levels. Thus, it appears that EOD duration is an accurate indicator of male status, which is under social and hormonal control. PMID- 11038293 TI - Partner preference of intact and ovariectomized female gray short-tailed opossums (Monodelphis domestica). AB - Intact, ovariectomized and ovariectomized estradiol (E)-treated female gray short tailed opossums were placed in a test situation in which they could choose between an intact and a castrated male. Intact females chose to visit intact males first and visited them more frequently and spent more time with intact than with castrated males. Ovariectomized (OVX) females did not show this preference for visiting intact males over castrates. When compared to OVX females with blank implants, OVX females with E implants spent less time with castrated males. Like intact females, OVX and OVX-E-treated females preferred to stay in close proximity to but not actually in the cage of intact rather than castrated males. To our knowledge, this is the first experimental study of partner preference and its relationship to hormonal condition in a female marsupial. PMID- 11038294 TI - Visible sympathetic activity as a social signal in Anolis carolinensis: changes in aggression and plasma catecholamines. AB - Darkening of postorbital skin in Anolis carolinensis occurs during stressful situations and is stimulated by sympathetic activation of beta(2)-adrenergic receptors via adrenal catecholamines. This eyespot forms more rapidly in dominant males during social interaction. Eyespot darkening (green to black) appears to function as a social signal communicating sympathetic activation and limiting aggressive interaction. To assess the value of the eyespot as a social signal, males were painted postorbitally with green, black, or red paint. Each male was exposed to a mirror following acclimation to the cage. The total number of aggressive displays toward the mirror image was greatest when eyespots were masked by green paint. In contrast, black or red artificial eyespots, regardless of size, inhibited biting behavior toward the mirror image. The most aggressive males, those who saw a reflected opponent with no eyespot (hidden with green paint), had significantly higher levels of all plasma catecholamines. These results suggest that A. carolinensis use information from the eyespot to assess their opponent's readiness to fight and thereby determine whether to be aggressive. Darkened eyespots are capable of inhibiting aggression, whereas aggressive displays from an opponent in the mirror without darkened eyespots do not. Darkened eyespots reflect rapid changes in plasma NE, DA, and Epi that may signal dominant social status. PMID- 11038295 TI - Insulin-like growth factor I receptor antagonism augments response to chemoradiation therapy in colon cancer cells. AB - BACKGROUND: Colorectal cancer remains one of the most prevalent malignancies in the United States. Improvement in local disease control is seen when 5 fluorouracil (5-FU) is used in combination with pelvic irradiation for rectal adenocarcinoma. The frequent overexpression of insulin-like growth factor I receptor (IGF-I-R) in rectal adenocarcinoma suggests that inhibition of the signal transduction pathway may be a novel approach to enhance tumor response. This investigation seeks to define the role of IGF-I-R antagonism, using monoclonal antibody alpha-IR3, in augmenting cytotoxicity to adjuvant chemoradiation therapy for adenocarcinoma of the rectum. MATERIALS AND METHODS: SW 480 colon cancer cells were cultured to semiconfluent conditions with dose titrations performed for 5-FU to determine that the IC(50) (inhibitory concentration of 50% of the cells) was 0.5 microg/ml. The IC(50) for 5-FU was reassessed in the presence of IGF-I. Experimental groups included colon cancer cells combined with 5-FU; 6-MeV external beam radiation (100-500 cGy); and alpha IR-3. RESULTS: The addition of 100 ng/ml IGF-I 1 h prior to 5-FU or radiation significantly blunted the expected cytotoxicity, resulting in a 10-fold increase in the IC(50) (from 0.5 to 5 microg/ml). Receptor antagonism using the monoclonal antibody alpha-IR-3 (100-400 ng/ml) produced a dose-dependent increase in cytotoxicity compared with 5-FU alone. The addition of radiation produced synergistic amplification of this response. CONCLUSIONS: IGF-I-R activation blocks the expected cytotoxic effects of 5-FU and external beam radiation. Receptor antagonism increased the cytotoxic response of chemoradiation therapy. These data suggest the utility of inhibiting IGF-I-R signal transduction in the treatment of rectal adenocarcinoma. PMID- 11038296 TI - Usefulness of an alcohol solution of N-duopropenide for the surgical antisepsis of the hands compared with handwashing with iodine-povidone and chlorhexidine: clinical essay. AB - BACKGROUND: The usual surgical antisepsis involves scrubbing the skin with antiseptic solutions. This procedure can damage the skin, with the subsequent risk of infection for the patient. There are several efficient and quick-acting antiseptic alcohol solutions that require no scrubbing. MATERIAL AND METHODS: We compare four alcohol solutions with the classic surgical handwashing products (chlorhexidine and iodine-povidone), in both in vitro (pigskin germ carriers) and in vivo studies. The latter (clinical essays) were done with healthy volunteers (crossed design) as well as with 154 surgical team members (Plastic Surgery or Traumatology), whose hand microbial flora were measured before and after scrubbing up and after surgery. RESULTS AND DISCUSSION: Because of its efficiency in the germ carrier, we chose a solution of N-duopropenide in 60 degrees alcohol with emollients for further comparison with the standard surgical scrub: 4% chlorhexidine and 7.5% iodine-povidone. The quantitative, semiquantitative, and qualitative results obtained with N-duopropenide without scrubbing were better in the healthy volunteers and surgical teams. This product reduced hand microorganisms by more than 2 log, and maintained the reduction for the entire study period. Four percent chlorhexidine initially reduced colonization more than 2 log but lost part of its effect over time during the surgical intervention. Last, 7.5% iodine-povidone reduced the germs by 1 log but at the end of surgery there were even more germs than before washing. CONCLUSION: Because of its efficacy, persistent effect, and skin protection, we advise that scrubbing with classic antiseptic solutions be replaced with gentle washing with an alcohol solution such as N-duopropenide in alcohol. PMID- 11038297 TI - Thermotolerance preserves endothelial vasomotor function during ischemia/reperfusion. AB - Ischemia/reperfusion (I/R) results in endothelial dysfunction, seen as loss of endothelium-dependent vasodilatation. In prolonged ischemia, this can result in marked vasospasm or no reflow in the microvasculature. Thermotolerance (T) attenuates I/R-induced microvascular injury. The aim of this study was to investigate the effect of thermotolerance on I/R-induced vasomotor changes and "no reflow." Sprague-Dawley rats were randomized into an ischemia/reperfusion group (I/R group) and a group in which thermotolerance (41 + 0.5 degrees C for 15 min 18 h prior to I/R) was induced (T + I/R group). IR injury was established by occlusion of the superior mesenteric and celiac vascular pedicle for 30 min, followed by 60 min of reperfusion. Vasomotor function [arteriolar constriction:dilatation (C:D) ratio] measured by response to acetylcholine (endothelium-dependent) and sodium nitroprusside (endothelium-independent) and "no-reflow" phenomenon were determined in mesenteric arterioles by intravital microscopy. Data are expressed as means +/- SEM and were analyzed using ANOVA and chi(2) test. I/R caused a significant decrease in endothelium-dependent vasodilatation (C:D = 1.37+/-0.31 in IR group vs. 2.06+/-0.20 in baseline, P<0.01) and no reflow in arterioles in 16 of 28 unheated rats. Endothelium independent dilatation was not altered by I/R. Thermotolerance attenuated this impairment of endothelium-dependent dilatation (P<0.01 vs. IR; C:D = 1.95+/-0.19) and reduced no-reflow phenomenon to 4 of 16 rats (P<0.05 vs. IR). This study demonstrated that thermotolerance preserves endothelial vasomotor function and markedly reduces "no reflow" in arterioles. PMID- 11038298 TI - Mechanisms of ischemic preconditioning in skeletal muscle. AB - BACKGROUND: Ischemic preconditioning (IP) (one or more cycles each consisting of a short period of ischemia and a short period of reperfusion, before the sustained ischemia) reduces ischemia-related organ damage in heart and skeletal muscle but the underlying mechanisms are not clear. This study was intended to assess the possible involvement of K(ATP) channels and of adenosine receptors in IP of skeletal muscle in a rat model of skeletal muscle ischemia. MATERIALS AND METHODS: Groups of 8-15 rats were given the following in vivo treatments: ischemia-reperfusion (I-R: 2.5 h tourniquet-induced ischemia of the right hindlimb, then 2 h reperfusion); IP (three cycles of 5 min ischemia, then 5 min reperfusion) before I-R; cromakalim and I-R; glibenclamide, cromakalim, and I-R; glibenclamide, IP, and I-R; [R]-N(6)-[1-methyl-2-phenylethyl]adenosine (R-PIA) and I-R; adenosine and I-R; and glibenclamide, IP, and I-R. Parameters of muscle function (postischemic maximal force, performance, contraction index, and force after 1 min of stimulation) were then assessed in vitro in the extensor digitorum longus muscle. RESULTS: Pretreatment with either IP or the K(ATP) channel opener cromakalim significantly improved postischemic muscle function. The protective effect of cromakalim was not seen when the K(ATP) channel blocker glibenclamide was added. Glibenclamide, however, did not block IP-induced protection. Pretreatment with the adenosine A(1) receptor agonist 8-(p-sulfophenyl) theophyllin (8-SPT) or with adenosine did not improve postischemic muscle function. The adenosine receptor agonist did not block IP-induced protection against ischemic damage. CONCLUSIONS: The results show significant improvements in postischemic skeletal muscle function after IP or cromakalim pretreatment but they do not support a role for K(ATP) channels or for adenosine receptors in IP of skeletal muscle. PMID- 11038299 TI - Plastic foil technique attenuates inflammation in mesenteric intravital microscopy. AB - BACKGROUND: Interpretation of intravital microscopic observations is complicated by the "inflammatory"-type response to the trauma inflicted on the tissue by the surgical preparation. The present study evaluates different experimental conditions for prolonged observations of the mesenteric microcirculation in the rat. METHODS: The mesentery was exteriorized through a median laparotomy and subjected to an organ bath or a modified plastic foil technique. Hemodynamic, metabolic, respiratory, and microcirculatory data were analyzed. RESULTS: In contrast to the plastic foil technique, which yielded stable baseline values over a 5-h observation period, venular velocity and wall shear rates decreased significantly in the organ bath technique, and leukocyte adhesion to the endothelium was significantly increased. Likewise, abdominal blood flow decreased significantly by 35% and base excess declined (-10.0+/-0.4 mmol/L) in the organ bath, with reduced pco(2) (26.4+/-2.5 mm Hg vs. 33.7+/-1.1 mm Hg in plastic foil technique) due to respiratory pH compensation. CONCLUSIONS: The plastic foil technique was found clearly superior to the organ bath technique for maintenance of stable baseline metabolic, hemodynamic, and microcirculatory conditions in mesenteric intravital microscopy. PMID- 11038300 TI - Glycation induced crosslinking of link proteins, in vivo and in vitro. AB - OBJECTIVE: Reducing sugars have the ability to crosslink proteins through creation of advanced glycosylated end products (AGE). In this study, we determined the ability of AGE to induce crosslinking of link proteins and aggrecan proteoglycans. METHODS: Aggrecan proteoglycans and link proteins were purified from adult human articular cartilage and from young bovine nasal cartilage for in vivo and in vitro studies, respectively. In vitro studies concerned incubation of aggrecan aggregates or link proteins with ribose under physiological conditions. After 30 days, aggregates were centrifuged dissociatively to obtain aggrecan monomers and link proteins. Aggrecan monomers were analyzed by immunoblot assay. Incubated link proteins were analyzed by SDS PAGE and Sephacryl-200 column chromatography. RESULTS: After extensive purification, adult human cartilage aggrecan continued to show the presence of link protein antigens by immunoblot analysis. Immunoblot analysis of purified aggrecan derived from ribose-treated aggregates also showed the presence of link protein antigens. Ribose treatment of link protein lead to polymerization that was confirmed by Sephacryl-200. CONCLUSIONS: These studies suggest that human link proteins tend to become crosslinked to aggrecan in adult cartilage. A likely cause of the crosslinking is formation of AGE due to reducing sugars. PMID- 11038301 TI - Microvasculature of small liver metastases in rats. AB - Neovascularization is important in the development of liver metastasis. We sought to define the origin and fine structure of the blood supply of small experimental liver metastases in rats using an injection replica method. Liver metastases were produced by intraportal inoculation of ascitic fluid containing AH60C hepatoma cells in male Donryu rats (n = 40). Intrahepatic microvasculature was studied by scanning electron microscopy and by stereomicroscopy of microvascular casts produced by perfusion via the abdominal aorta or portal vein 7 days following tumor inoculation. Intrahepatic microvasculature in rats without liver metastases (n = 10) also was studied by scanning electron microscopy. In the normal liver, branches of the hepatic artery typically terminated in the peribiliary plexus and less frequently led to sinusoids and terminal portal veins. In 69 metastatic tumors ranging from 269 to 1875 microm in diameter, arterially perfused metastatic tumors larger than 300 microm showed newly developed neovascularization. Portally perfusion of metastatic tumors did not visualize neovascularization irrespective of tumor size. At the periphery of metastases, tumor vessels disclosed by arterial perfusion most often communicated with the peribiliary plexus and less frequently with terminal arterioles. Metastatic liver tumors as small as 300 microm in diameter receive their main blood supply from the hepatic artery but not from the portal vein, and tumor vessels more often are derived from the arterially filled peribiliary plexus rather than from terminal arterioles. PMID- 11038302 TI - ATP induces c-fos expression in C6 glioma cells by activation of P(2Y) receptors. AB - BACKGROUND: Extracellular ATP functions in the enteric nervous system as a neurotransmitter, and recent evidence suggests ATP may regulate development through effects on cellular proliferation. METHODS: The action of ATP at purinoceptors and the role of second messenger pathways in c-fos mRNA expression in C6 glioma cells were investigated using the techniques of Northern and Western blotting. RESULTS: Treatment of C6 cells with ATP caused a time- and dose dependent increase in c-fos expression. The rank order of agonist potency was ATP = ADP > gammasATP > alphabetaATP > betagammaATP > AMP = UTP. The ATP-induced c fos increment was inhibited by three P(2Y) receptor antagonists-suramin, reactive blue, and DIDS-by 99+/-3, 89+/-7, and 61+/-14%, respectively. The ATP-stimulated c-fos expression was attenuated by phospholipase C inhibitor (U73122), protein kinase C (PKC) down-regulation (4alpha-phorbol 12-myristate 13-acetate and chelerythrine), mitogen-activated protein (MAP) kinase inhibition (apigenin), an inhibitor of MAP kinase kinase (PD98059), down-regulation of adenylate cyclase (SQ22536), and inhibition of type II protein kinase A (8-(4 chlorophenylthio)adenosine-3',5'-cyclic monophosphorothioate), but was not affected by inhibition of type I protein kinase A (8-bromoadenosine-3',5'-cyclic monophosphorothioate) and inhibitors of calmodulin kinase (KN93 and KN62). Phosphorylated MAP kinase was increased in cells exposed to ATP. This effect was suppressed by chelerythrine. CONCLUSIONS: These studies demonstrate that ATP induced c-fos mRNA expression is under multifactorial regulation. PMID- 11038303 TI - Expression of cell-cycle regulators p27 and cyclin E correlates with survival in gastric carcinoma patients. AB - BACKGROUND: Cell-cycle control is important in carcinogenesis and cancer progression. p27 and cyclin E are cell-cycle regulators, which control the G1-S phase transition. Recently, these two factors were found to be affected in many human cancers. The aim of the study was to examine the expression of p27 and cyclin E in gastric cancer and to evaluate their prognostic implication. MATERIALS AND METHODS: Paraffin blocks of 56 samples of advanced gastric cancer, 15 samples of early gastric cancer, and 17 samples of normal gastric mucosa were studied. Expression of p27 and cyclin E was analyzed by immunohistochemistry. The relationship between the expression and clinicopathological data was examined. RESULTS: Expression of p27 was reduced in 89% of advanced cancer samples, 44% of early cancer samples, and 12% of normal mucosa samples (P<0.0001). Among the cancers, reduced expression of p27 was associated with a large tumor size, increased cancer invasion, nodal metastases, and the presence of residual tumor after operation. No significant difference in cyclin E expression was found. Kaplan-Meier plots of survival showed tumors with low p27 were associated with poorer survival than those with high p27 expression (RR, 5.3; CI = 1.6-17.4; P = 0.005). Tumors with low p27 and high cyclin E expression were associated with the highest mortality expression (RR, 9.8; CI = 1.2-80; P = 0.03). CONCLUSIONS: Gastric cancer with low expression of p27 is associated with aggressive characteristics and a poorer outcome. PMID- 11038304 TI - Interdependence of adenosine and nitric oxide in hepato-splanchnic circulation during sepsis. AB - BACKGROUND: We tested the hypothesis that some of the maintenance of resting, regional hepato-splanchnic perfusion that is mediated by endogenous adenosine (ADO) during sepsis is interdependent with nitric oxide (NO). MATERIALS AND METHODS: Twenty-four hours after sepsis/sham induction, rats were divided into two groups. Group 1 received a 10-min infusion of the ADO antagonist 8 sulfophenyltheophylline (8-SPT; 0.9 mg/kg x min), followed by 10 min of 8-SPT plus L-NAME (0.5 mg/kg x min). Group 2 received L-NAME first followed by 8-SPT in the presence of L-NAME (all groups: n = 6-10). Hemodynamic and regional hepato splanchnic blood flow measurements were made prior to infusions, 10 min after initiation of each single agent infusion, and again after double agent infusion. RESULTS: Twenty-four hours after sepsis hepato-splanchnic blood flow was significantly elevated, compared to nonseptic rats. Both ADO receptor blockade alone and NOS inhibition alone decreased total hepato-splanchnic blood flow to nonseptic values. Decreases in small intestinal and cecal blood flow accounted for the majority of this decrease, but decreased hepatic arterial perfusion contributed as well. No further alterations were seen when 8-SPT was infused in the presence of L-NAME, nor when L-NAME was infused in the presence of 8-SPT. CONCLUSIONS: These data indicate that there is significant interdependence of endogenous NO and ADO in maintaining resting small bowel, cecal, and hepatic arterial perfusion during sepsis. The lack of responses in other regions of the hepato-splanchnic circulation demonstrate regional specificity of this ADO-NO interdependence. PMID- 11038305 TI - Biochemical changes in transplanted rat liver stored in University of Wisconsin and Euro-Collins solutions. AB - BACKGROUND: Liver ischemia/reperfusion injury is a severe problem in transplantation, and preservation solutions could be critical for liver viability. The aim of our study was to evaluate the cytosolic and mitochondrial glutathione levels, the glyoxalase II activity, and the mitochondrial hydroperoxide contents of livers stored in different preservation solutions for 7 or 24 h and after transplantation. MATERIALS AND METHODS: Orthotopic liver transplantation was performed without reconstruction of the hepatic artery. The livers were stored at 4 degrees C for 7 or 24 h in University of Wisconsin or Euro-Collins solutions. Portions of livers before and after transplantation were homogenized and mitochondria isolated. RESULTS: Cytosolic glutathione levels were decreased in all stored livers and after transplantation. In livers stored with University of Wisconsin solution, mitochondrial glutathione was unchanged during cold storage and no significant decrease has been found after reperfusion, whereas in livers stored in Euro-Collins solution, mitochondrial glutathione was decreased and a further significant decrease was found 30 min after reperfusion. Mitochondrial hydroperoxides were higher in livers stored in Euro-Collins solution than in University of Wisconsin solution after 30 min of reperfusion. Mitochondrial glyoxalase II did not show any change by reperfusion. CONCLUSION: We have demonstrated that rat liver stored in Euro-Collins solution suffered a severe depletion of mitochondrial GSH and a concomitant increase of hydroperoxides. The data obtained suggested that the livers stored with University of Wisconsin solution were probably less prone to ischemia/reperfusion injury after liver transplantation. PMID- 11038306 TI - Schistosoma mansoni: adhesion of mannan-binding lectin to surface glycoproteins of cercariae and adult worms. AB - Schistosoma mansoni is a blood-dwelling trematode which can persist for several years in the vessels of the human host. The schistosomal surface has been extensively characterized by lectin binding studies, revealing the carbohydrate composition of the worm's tegument. Using fluorescent and scanning electron microscopy we demonstrate that the surface carbohydrates of cercariae and adult worms are the binding ligands for mannanbinding lectin (MBL), a serum protein that is part of the innate immune system. An in vitro complement activation assay with C1q-deficient complement suggests that MBL, in association with the serine proteases MASP-1 and MASP-2, is capable of fixing complement components on the schistosomal tegument and activating the complement cascade via the "MBL pathway." MBL is constitutively expressed by hepatocytes and present in the blood at a stable level. Since it is also a weak acute-phase protein and therefore upregulated in an acute-phase response we investigated the serum MBL levels in patients infected with Schistosoma sp. and in healthy control persons. An enzyme linked immunosorbent assay indicated no differences between the two groups. Although our results suggest an involvement of MBL activated complement in vitro, its role in vivo remains to be clarified. PMID- 11038307 TI - Uptake and interconversion of fluorescent lipid analogs in the protozoan parasite, Perkinsus marinus, of the oyster, Crassostrea virginica. AB - Uptake, distribution, and interconversion of fluorescent lipid analogs (phosphatidylcholine, PC; cholesteryl ester, CHE; phosphatidylethanolamine, PE; palmitic acid, C16; sphingomyelin, SM) by the two life stages, meront and prezoosporangium, of the oyster protozoan parasite, Perkinsus marinus, were investigated. Class composition of these two life stages and lipid contents in meront cells were also examined. Both meronts and prezoosporangia incorporated and modified fluorescent lipids from the medium, but their metabolic modes differ to some extent. Results revealed that among the tested analogs, neutral lipid components (CHE and C16) were incorporated to a greater degree than the phospholipids (PC, PE, and SM). HPLC analysis of meront lipids showed that while the majority of the incorporated PC, CHE, and PE remained as parent compounds, most of the incorporated C16 was in triacylglycerol (TAG) and SM was in ceramide and free fatty acids. The cellular distribution of fluorescent labels varied with lipid analogs and the extent of their metabolism by the parasite. Fluorescence distribution was primarily in cytoplasmic lipid droplets of both life stages after 24 h incubation with PC. After 24 h incubation with SM, fluorescence appeared in the membrane and cytosol. Total lipid contents in meront cultures increased during proliferation and TAG accounted for most of the increased total lipids. Since total lipid content per meront cell did not increase until the day of culture termination, the lipid increase in the meront culture was mainly a result of increased cell numbers. Both life stages contain relatively high levels of phospholipids, 53.8% in 8-day-old meronts and 39.4% in prezoosporangia. PC was the predominant phospholipid. PMID- 11038308 TI - Development of a serum-free system for the in vitro cultivation of Brugia malayi infective-stage larvae. AB - Over the past several years, numerous attempts have been made to culture the infective-stage (L3) larvae of the human filarial parasite Brugia malayi in an in vitro system that promotes molting to the fourth larval stage (L4). Although there have been reports in the literature of successful L3 to L4 development in vitro, all of these systems have required serum supplementation. The complexity of serum as a culture supplement has made reproducibility of results and identification of specific factors necessary for L3 development problematic. We have developed a serum-free in vitro system consisting of RPMI 1640 supplemented with one of three fatty acids (arachidonic, linoleic, or linolenic) that supports consistent and reproducible molting to the fourth larval stage in the presence of a basidiomycetous yeast, Rhodotorula minuta. Coculture with this yeast, initially isolated as a culture contaminant, is required for successful molting. In serum free cultures lacking R. minuta, L3 larvae survive for upward of 2 weeks, but do not molt successfully. The L4 larvae generated in cultures containing R. minuta are well formed, as seen by light and electron microscopy, and are capable of further development upon transfer to a permissive host. This culture system is inexpensive and easily reproducible, thus making it a useful tool for studying the requirements for the development of B. malayi L3. PMID- 11038309 TI - Tetracycline inhibits development of the infective-stage larvae of filarial nematodes in vitro. AB - In recent years, studies have linked tetracycline treatment of filaria-infected animals with reduced adult worm burdens and decreased levels of microfilaremia. These observations are believed to be attributable to clearance of Wolbachia, intracellular rickettsial-like organisms found within filarial tissues. Although maximal worm reductions were observed when treatment was initiated early in infection, it is not known whether tetracycline inhibits development of infective stage larvae. To address this issue, we studied the effect of tetracycline on three different species of filarial nematodes, Brugia malayi, Brugia pahangi, and Dirofilaria immitis, in a serumfree in vitro system supporting molting to the fourth larval stage. Tetracycline was capable of inhibiting L3 to L4 molting within a dosage range similar to that reported for susceptible rickettsial organisms. However, Wolbachia DNA could still be detected in nematodes from tetracycline-treated cultures. In addition, three other antibiotics with anti rickettsial and anti-chlamydial activity (chloramphenicol, erythromycin, and ciprofloxacin) failed to inhibit L3 to L4 molting. Although tetracycline is capable of completely blocking molting of infective-stage larvae, it remains possible that this effect is due to pharmacological activities unrelated to its anti-rickettsial functions. PMID- 11038311 TI - Leishmania mexicana mexicana: genetic heterogeneity of mexican isolates revealed by restriction length polymorphism analysis of kinetoplast DNA. AB - Leishmania mexicana mexicana isolates from 23 patients with localized, diffuse, and an atypical "pseudodiffuse" form of cutaneous leishmaniasis were obtained in various endemic regions of Mexico. Restriction fragment length polymorphism analysis of kinetoplast DNA was done with nine different endonucleases in addition to an in vitro growth pattern analysis. We found that the 23 L. mexicana mexicana isolates could be consistently classified into six groups, according to the endonuclease digestion patterns obtained with HaeIII, HpaII, and MseI. Whereas localized cutaneous leishmaniasis isolates could have any of five patterns, diffuse cutaneous leishmaniasis showed only two patterns and pseudodiffuse cutaneous leishmaniasis consistently showed only one pattern. Thus, a clear correlation among digestion pattern, clinical disease, and geographical localization was obtained for the pseudodiffuse cutaneous leishmaniasis group. Additionally, the L. mexicana mexicana isolates could be differentiated into fast and slow-growing groups. Diffuse cutaneous leishmaniasis isolates were found to be fast growing, whereas localized cutaneous leishmaniasis isolates fell into both categories. In contrast, all pseudo diffuse cutaneous leishmaniasis isolates were slow growing. Here we report the first study in which distinct and persistent genotypic characteristics of kinetoplast DNA heterogeneity within the L. mexicana mexicana species could be directly correlated with clinical disease and its growth behavior, suggesting that a distinctive restriction pattern could have important biological implications. Additionally, this study sheds new light on the biological significance of parasite kinetoplast DNA, since the heterogeneity seems not to be random but to form a distinct pattern. PMID- 11038310 TI - A molecular tool for species identification and benzimidazole resistance diagnosis in larval communities of small ruminant parasites. AB - This report describes a molecular method for determining in a first step the generic composition of a nematode community and in a second step, the resistance of each species to benzimidazole (BZ). We first established a polymerase chain reaction (PCR) linked to a restriction fragment length polymorphism strategy using the isotype 1 beta-tubulin gene. This method overcame the limitations of morphological identification of larval stages of trichostrongylid nematode species. Geographically distant isolates from the three main gastrointestinal species in temperate zones, Teladorsagia circumcincta, Haemonchus contortus, and Trichostrongylus colubriformis, were distinguished using this method. We then used an allele-specific PCR (AS-PCR) to detect mutations of residue 200 of the beta-tubulin, which is implicated in BZ resistance. The sequences of several samples confirmed the BZ-resistance genotype determined by AS-PCR. The ability to process large numbers of samples simultaneously makes this PCR-based strategy particularly suitable for epidemiological studies. It may also be useful for monitoring the emergence of resistant alleles in nematode communities. PMID- 11038312 TI - Entamoeba histolytica: rapid detection of indian isolates by cysteine proteinase gene-specific polymerase chain reaction. AB - Amoebiasis, caused by Entamoeba histolytica, is still one of the major problems for developing countries like India. Early detection of the parasite is a must for its prevention and control. In this study, PCR analysis of the cysteine proteinase gene from clinical isolates of symptomatic intestinal and amoebic liver abscess (ALA) cases has been compared with the stool microscopy, serology, and ultrasonography methods. The clinical isolates negative for E. histolytica by stool microscopy demonstrated the presence of the cysteine proteinase gene by PCR amplification. Also the gene copy number was increased in ALA samples compared with intestinal cases. Hence an accurate, early, and easier detection was possible by cysteine proteinase gene amplification directly from the clinical samples. PMID- 11038313 TI - Entamoeba invadens: protein kinase C inhibitors block the growth and encystation. PMID- 11038314 TI - Efficacy estimates from parasite count data that include zero counts. AB - Because of the positive skewness of parasite distributions and the greater constancy of percentage of response of therapy in animal populations, parasite count data are conventionally transformed logarithmically before combining results from different animals, either all controls or all treated. Observations of zero counts raise difficulties, since the logarithm of zero is not useful. In this study, several types of zero count adjustments are compared. Two systems for assigning values to zero counts were considered: a fixed system, which assigns the same value to all zero counts regardless of the proportion of such counts in a treatment group, and a variable system, which replaces zero counts with a value based on the proportion of zero counts in the group. The values assigned by either system are then adjusted to reflect aliquot size. An evaluation was performed by using 32 compound Poisson lognormal distributions, three sample sizes, and three representatives of each zero count adjustment system. The Poisson lognormal distribution provides a convenient method with which to provide variability greater than Poisson. Expected values of the sample estimate of the (known) population mean were calculated for each of the 576 combinations of these factors, and the bias associated with each combination was derived. The bias associated with the three representatives of the variable adjustment system was similar. The variable adjustment system had a lower overall bias than any representatives of the fixed adjustment system. PMID- 11038315 TI - Blastocystis hominis: A simplified, high-efficiency method for clonal growth on solid agar. AB - Colony growth of protozoan parasites in agar can be useful for axenization, cloning, and viability studies. This is usually achieved with the pour plate method, for which the parasite colonies are situated within the agar. This technique has been described for Giardia intestinalis, Trichomonas vaginalis, and Entamoeba and Blastocystis species. Extracting such colonies can be laborious. It would be especially useful if parasites could be grown on agar as colonies. These colonies, being exposed on the agar surface, could be conveniently isolated for further investigation. In this study, we report the successful culture of B. hominis cells as colonies on solid agar. Colonies were enumerated and the efficiency of plating was determined. It was observed that B. hominis could be easily cultured on agar as clones. The colonies were dome-shaped and mucoid and could grow to 3 mm in diameter. Flow cytometric analyses revealed that parasite colonies remained viable for up to 2 weeks. Viable colonies were conveniently expanded in liquid or solid media. Scanning electron microscopy revealed that each colony consists of two regions; a dome-shaped, central core region and a flattened, peripheral region. Older colonies possessed numerous strand-like surface coat projections. This study provides the first report of clonal growth of B. hominis on agar and a simple, effective method for cloning and expansion of B. hominis cells. PMID- 11038316 TI - Trypanosoma brucei: four tandemly linked genes for fatty acyl-CoA synthetases. AB - We have cloned four acyl CoA synthetase (ACS) genes from Trypanosoma brucei strain 927. Each of these genes encodes a polypeptide about 78 kDa in size and all four contain the "ACS signature motif." Sequence alignments indicate that these proteins are 46%-95% identical in amino acid sequence. Interestingly, three of them share almost identical C-termini (about 215 amino acid residues). Southern blots suggest that these genes are present in a single copy, and Northern blots reveal that all four are expressed in both bloodstream and procyclic trypanosomes. PMID- 11038317 TI - Trypanosoma cruzi: cloning and characterization of a RAB7 gene. AB - The small monomeric GTP-binding proteins of the RAB subfamily are key regulatory elements of the machinery that controls membrane traffic in eukaryotic cells. These proteins have been localized to many different intracellular organelles, on both endocytic and exocytic compartments, suggesting that each step of vesicular traffic can involve a specific RAB protein. The presence of conserved amino acid domains in these proteins has allowed the cloning of their genes from several organisms, including yeast, plants, humans, and parasites. In this work we describe the identification, cloning, and characterization of a RAB7 gene homologue in Trypanosoma cruzi (TcRAB7). Our data indicate that this gene is present as a single copy in the T. cruzi genome, located on a 2.25-Mb chromosomal DNA. TcRAB7 is expressed in T. cruzi epimastigotes, metacyclic trypomastigotes, and spheromastigotes. We established transformed cell lines that express two versions of an epitope-tagged TcRAB7 protein: one wild type (pTAG) and one deleted at the C-terminal cysteines (pDeltaCXC). Wild-type TcRAB7 protein (pTAG) appears to be localized exclusively in the membrane fraction, while the mutated TcRAB7 protein (pDeltaCXC) loses the ability to associate with the membrane, showing only cytosolic localization. Also, we produced the recombinant TcRAB7 protein and demonstrated that it binds GTP. The identification of exo- and endocytic machinery components in T. cruzi and their function would provide specific markers of these subcellular compartments, thereby unveiling important aspects of vesicular traffic in this parasite. PMID- 11038318 TI - A 23-kDa recombinant antigen of Cryptosporidium parvum induces a cellular immune response on in vitro stimulated spleen and mesenteric lymph node cells from infected mice. AB - In the present study, we focused on a 23-kDa antigen, Cp23, which has been shown to be a major target of humoral immune responses in Cryptosporidium parvum infections and is present in both the sporozoite and merozoite stages. Recombinant Cp23 antigen was shown to stimulate a specific proliferative response by splenocytes and mesenteric lymph node cells from infected interferon gamma knockout BALB/c mice. Cp23 stimulation also induced TNF-alpha, IL-2, and IL-5 mRNA production by spleen cells from infected animals. In contrast, IL-12 mRNA was decreased by Cp23 stimulation compared with unstimulated splenocytes. These data suggest that, as with humoral responses, Cp23 is an important target of cellular immune responses in experimental C. parvum infections. The potential role of this antigen in conferring protective immunity is also discussed. PMID- 11038319 TI - Giardia lamblia in Mongolian gerbils: characteristics of infection using different human isolates. PMID- 11038320 TI - Schistosoma haematobium: population genetics of S. haematobium by direct measurement of parasite diversity using RAPD-PCR. PMID- 11038321 TI - Entamoeba histolytica: bacterial expression of a human monoclonal antibody which inhibits in vitro adherence of trophozoites. PMID- 11038322 TI - Entamoeba histolytica: deletion of the GPI anchor signal sequence on the Gal/GalNAc lectin light subunit prevents its assembly into the lectin heterodimer. AB - Adherence and cytotoxicity of Entamoeba histolytica require the function of a heterodimeric galactose and N-acetylgalactosamine (Gal/GalNAc)-specific lectin. The lectin heavy subunit (Hgl) contains a carbohydrate recognition domain and mediates inside-out cell signaling via its cytoplasmic tail. The function of the lectin light subunit (Lgl) is unknown. The lectin has a unique mechanism of membrane association: Hgl is transmembrane but Lgl is glycosylphosphatidylinositol (GPI) anchored. The role of the GPI anchor signal sequence in heterodimer assembly was tested. Epitope-tagged Lgl with or without the GPI anchor addition signal was expressed in E. histolytica trophozoites. Tagged Lgl did not assemble with Hgl into a lectin heterodimer in the absence of the GPI addition signal. Consistent with previous results that only the Hgl subunit mediates adherence, the monomeric Lgl without the GPI anchor signal lacked Gal/GalNAc-binding activity. PMID- 11038323 TI - Apparently normal ovarian differentiation in a prepubertal girl with transcriptionally inactive steroidogenic factor 1 (NR5A1/SF-1) and adrenocortical insufficiency. AB - Steroidogenic factor 1 (NR5A1/SF-1) plays an essential role in the development of the hypothalamic-pituitary-adrenal and hypothalamic-pituitary-gonadal axes, controlling expression of their many important genes. The recent description of a 46,XY patient bearing a mutation in the NR5A1 gene, causing male pseudohermaphroditism and adrenal failure, demonstrated the crucial role of SF-1 in male gonadal differentiation. The role of SF-1 in human ovarian development was, until now, unknown. We describe a phenotypically and genotypically normal girl, with signs and symptoms of adrenal insufficiency and no apparent defect in ovarian maturation, bearing a heterozygote G-->T transversion in exon 4 of the NR5A1 gene that leads to the missense R255L in the SF-1 protein. The exchange does not interfere with protein translation and stability. Consistent with the clinical picture, R255L is transcriptionally inactive and has no dominant negative activity. The inability of the mutant (MUT) NR5A1/SF-1 to bind canonical DNA sequences might offer a possible explanation for the failure of the mutant protein to transactivate target genes. This is the first report of a mutation in the NR5A1 gene in a genotypically female patient, and it suggests that NR5A1/SF-1 is not necessary for female gonadal development, confirming the crucial role of NR5A1/SF-1 in adrenal gland formation in both sexes. PMID- 11038324 TI - A novel homoplasmic mutation in mtDNA with a single evolutionary origin as a risk factor for cardiomyopathy. AB - To clarify the relationship between variation in mtDNA and the development of cardiomyopathy (CM), the complete sequences of mtDNAs of two brothers with dilated CM were compared with those of 181 patients who had CM and with those of 168 control subjects. Five patients with CM shared a novel homoplasmic point mutation (G12192A tRNA(His)), and all of them demonstrated the evolutionarily related D-loop sequence. The results suggest that this novel mutation originated from the same ancestor and that its presence strongly predisposes carriers to CM. PMID- 11038325 TI - Partial paternal uniparental disomy of chromosome 6 in an infant with neonatal diabetes, macroglossia, and craniofacial abnormalities. AB - Neonatal diabetes, which can be transient or permanent, is defined as hyperglycemia that presents within the first month of life and requires insulin therapy. Transient neonatal diabetes mellitus has been associated with abnormalities of the paternally inherited copy of chromosome 6, including duplications of a portion of the long arm of chromosome 6 and uniparental disomy, implicating overexpression of an imprinted gene in this disorder. To date, all patients with transient neonatal diabetes mellitus and uniparental disomy have had complete paternal isodisomy. We describe a patient with neonatal diabetes, macroglossia, and craniofacial abnormalities, with partial paternal uniparental disomy of chromosome 6 involving the distal portion of 6q, from 6q24-qter. This observation demonstrates that mitotic recombination of chromosome 6 can also give rise to uniparental disomy and neonatal diabetes, a situation similar to that observed in Beckwith-Wiedemann syndrome, another imprinted disorder. This finding has clinical implications, since somatic mosaicism for uniparental disomy of chromosome 6 should also be considered in patients with transient neonatal diabetes mellitus. PMID- 11038326 TI - Genetic susceptibility to thrombosis and its relationship to physiological risk factors: the GAIT study. Genetic Analysis of Idiopathic Thrombophilia. AB - Although there are a number of well-characterized genetic defects that lead to increased risk of thrombosis, little information is available on the relative importance of genetic factors in thrombosis risk in the general population. We performed a family-based study of the genetics of thrombosis in the Spanish population to assess the heritability of thrombosis and to identify the joint actions of genes on thrombosis risk and related quantitative hemostasis phenotypes. We examined 398 individuals in 21 extended pedigrees. Twelve pedigrees were ascertained through a proband with idiopathic thrombosis, and the remaining pedigrees were randomly ascertained. The heritability of thrombosis liability and the genetic correlations between thrombosis and each of the quantitative risk factors were estimated by means of a novel variance component method that used a multivariate threshold model. More than 60% of the variation in susceptibility to common thrombosis is attributable to genetic factors. Several quantitative risk factors exhibited significant genetic correlations with thrombosis, indicating that some of the genes that influence quantitative variation in these physiological correlates also influence the risk of thrombosis. Traits that exhibited significant genetic correlations with thrombosis included levels of several coagulation factors (factors VII, VIII, IX, XI, XII, and von Willebrand), tissue plasminogen activator, homocysteine, and the activated protein C ratio. This is the first study that quantifies the genetic component of susceptibility to common thrombosis. The high heritability of thrombosis risk and the significant genetic correlations between thrombosis and related risk factors suggest that the exploitation of correlated quantitative phenotypes will aid the search for susceptibility genes. PMID- 11038327 TI - The economic value of bioinformation. PMID- 11038328 TI - Sister-scanning: a Monte Carlo procedure for assessing signals in recombinant sequences. AB - MOTIVATION: To devise a method that, unlike available methods, directly measures variations in phylogenetic signals in gene sequences that result from recombination, tests the significance of the signal variations and distinguishes misleading signals. RESULTS: We have developed a method, that we call 'sister scanning', for assessing phylogenetic and compositional signals in the various patterns of identity that occur between four nucleotide sequences. A Monte Carlo randomization is done for all columns (positions) within a window and Z-scores are obtained for four real sequences or three real sequences with an outlier that is also randomized. The usefulness of the approach is demonstrated using tobamovirus and luteovirus sequences. Contradictory phylogenetic signals were distinguished in both datasets, as were regions of sequence that contained no clear signal or potentially misleading signals related to compositional similarities. In the tobamovirus dataset, contradictory phylogenetic signals were separated by coding sequences up to a kilobase long that contained no clear signal. Our re-analysis of this dataset using sister-scanning also yielded the first evidence known to us of an inter-species recombination site within a viral RNA-dependent RNA polymerase gene together with evidence of an unusual pattern of conservation in the three codon positions. PMID- 11038329 TI - Secondary structure alone is generally not statistically significant for the detection of noncoding RNAs. AB - MOTIVATION: Several results in the literature suggest that biologically interesting RNAs have secondary structures that are more stable than expected by chance. Based on these observations, we developed a scanning algorithm for detecting noncoding RNA genes in genome sequences, using a fully probabilistic version of the Zuker minimum-energy folding algorithm. RESULTS: Preliminary results were encouraging, but certain anomalies led us to do a carefully controlled investigation of this class of methods. Ultimately, our results argue that for the probabilistic model there is indeed a statistical effect, but it comes mostly from local base-composition bias and not from RNA secondary structure. For the thermodynamic implementation (which evaluates statistical significance by doing Monte Carlo shuffling in fixed-length sequence windows, thus eliminating the base-composition effect) the signals for noncoding RNAs are still usually indistinguishable from noise, especially when certain statistical artifacts resulting from local base-composition inhomogeneity are taken into account. We conclude that although a distinct, stable secondary structure is undoubtedly important in most noncoding RNAs, the stability of most noncoding RNA secondary structures is not sufficiently different from the predicted stability of a random sequence to be useful as a general genefinding approach. PMID- 11038330 TI - Clustalnet: the joining of Clustal and CORBA. AB - MOTIVATION: Performing sequence alignment operations from a different program than the original sequence alignment code, and/or through a network connection, is often required. Interactive alignment editors and large-scale biological data analysis are common examples where such a flexibility is important. Interoperability between the alignment engine and the client should be obtained regardless of the architectures and programming languages of the server and client. RESULTS: Clustalnet, a Clustal alignment CORBA server is described, which was developed on the basis of Clustalw. This server brings the robustness of the algorithms and implementations of Clustal to a new level of reuse. A Clustalnet server object can be accessed from a program, transparently through the network. We present interfaces to perform the alignment operations and to control these operations via immutable contexts. The interfaces that select the contexts do not depend on the nature of the operation to be performed, making the design modular. The IDL interfaces presented here are not specific to Clustal and can be implemented on top of different sequence alignment algorithm implementations. PMID- 11038331 TI - Domain size distributions can predict domain boundaries. AB - MOTIVATION: The sizes of protein domains observed in the 3D-structure database follow a surprisingly narrow distribution. Structural domains are furthermore formed from a single-chain continuous segment in over 80% of instances. These observations imply that some choices of domain boundaries on an otherwise uncharacterized sequence are more likely than others, based solely on the size and segment number of predicted domains. This property might be used to guess the locations of protein domain boundaries. RESULTS: To test this possibility we enumerate putative domain boundaries and calculate their relative likelihood under a probability model that considers only the size and segment number of predicted domains. We ask, in a cross-validated test using sequences with known 3D structure, whether the most likely guesses agree with the observed domain structure. We find that domain boundary predictions are surprisingly successful for sequences up to 400 residues long and that guessing domain boundaries in this way can improve the sensitivity of threading analysis. PMID- 11038333 TI - Object-oriented parsing of biological databases with Python. AB - MOTIVATION: While database activities in the biological area are increasing rapidly, rather little is done in the area of parsing them in a simple and object oriented way. RESULTS: We present here an elegant, simple yet powerful way of parsing biological flat-file databases. We have taken EMBL, SWISSPROT and GENBANK as examples. EMBL and SWISS-PROT do not differ much in the format structure. GENBANK has a very different format structure than EMBL and SWISS-PROT. Extracting the desired fields in an entry (for example a sub-sequence with an associated feature) for later analysis is a constant need in the biological sequence-analysis community: this is illustrated with tools to make new splice site databases. The interface to the parser is abstract in the sense that the access to all the databases is independent from their different formats, since parsing instructions are hidden. PMID- 11038332 TI - Using traveling salesman problem algorithms for evolutionary tree construction. AB - MOTIVATION: The construction of evolutionary trees is one of the major problems in computational biology, mainly due to its complexity. RESULTS: We present a new tree construction method that constructs a tree with minimum score for a given set of sequences, where the score is the amount of evolution measured in PAM distances. To do this, the problem of tree construction is reduced to the Traveling Salesman Problem (TSP). The input for the TSP algorithm are the pairwise distances of the sequences and the output is a circular tour through the optimal, unknown tree plus the minimum score of the tree. The circular order and the score can be used to construct the topology of the optimal tree. Our method can be used for any scoring function that correlates to the amount of changes along the branches of an evolutionary tree, for instance it could also be used for parsimony scores, but it cannot be used for least squares fit of distances. A TSP solution reduces the space of all possible trees to 2n. Using this order, we can guarantee that we reconstruct a correct evolutionary tree if the absolute value of the error for each distance measurement is smaller than f2.gif" BORDER="0">, where f3.gif" BORDER="0">is the length of the shortest edge in the tree. For data sets with large errors, a dynamic programming approach is used to reconstruct the tree. Finally simulations and experiments with real data are shown. PMID- 11038334 TI - Digital reviews in molecular biology: approaches to structured digital publication. AB - MOTIVATION: It is widely appreciated that it is no longer possible for biomedical research scientists to keep up with as much of what is published in their field as they ought. One solution to this problem is to increase the efficiency of information use by moving away from the classical browsing model for scientific information dissemination towards an information on demand model which would allow researchers to access information quickly and efficiently only as they need it. The most common approach to this goal has been to use information retrieval technology to improve access to text databases of biomedical information. We are interested in exploring an alternative; encoding this information for storage in structured databases for efficient retrieval. RESULTS: Two small databases described here are test beds for development of structured digital publication software; the Tumor Gene Database, containing information about genes which are the sites for cancer-causing mutations, and the Mammary Transgene Database, containing information about expression of transgenes in agriculturally important animals. Both have been successfully searched by users and edited by curators via the World Wide Web. PMID- 11038335 TI - eSAGE: managing and analysing data generated with serial analysis of gene expression (SAGE). AB - SUMMARY: eSAGE is a comprehensive set of software tools for managing and analysing data generated with Serial Analysis of Gene Expression (SAGE). PMID- 11038336 TI - DNAssist: the integrated editing and analysis of molecular biology sequences in windows. AB - MOTIVATION: The programs currently available for the analysis of nucleic acid and protein sequences suffer from a variety of problems: Web-based programs often require inconvenient reformatting of sequences when proceeding from one analysis to the next, and commercial-console-based programs are cost prohibitive. Here, we report the development of DNASSIST:, an inexpensive, multiple-document, interface program for the fully integrated editing and analysis of nucleic acid and protein sequences in the familiar environment of Microsoft Windows. PMID- 11038337 TI - A cross-comparison of a large dataset of genes. AB - SUMMARY: We make available a large cross-comparison for 16 of the completely sequenced genomes and additional eukaryotic genes. The alignments were performed at the protein level using liberal similarity bounds in order to capture as many significant alignments as possible. This dataset will be updated as new genomes become available. PMID- 11038338 TI - NAIL-Network Analysis Interface for Linking HMMER results. AB - SUMMARY: Network Analysis Interface for Linking HMMER results (NAIL) is a web based tool for the analysis of results from a HMMER protein database-search. NAIL facilitates the selection of protein hits and the creation of an alignment, which can be used for a new sequence similarity search. PMID- 11038339 TI - SChiSM: creating interactive web page annotations of molecular structure models using chime. AB - SUMMARY: SChiSM is a program for creating World Wide Web (WWW) pages that include embedded interactive molecular models using the browser plug-in, CHIME:, for visualization. The program works with Netscape 4.x and Internet Explorer 5 browsers to facilitate Chime/Rasmol scripting control of a molecular display. PMID- 11038340 TI - Visualizing large hierarchical clusters in hyperbolic space. AB - SUMMARY: HyperTree is an application to visualize and navigate large trees in hyperbolic space. It includes color-coding, search mechanisms and navigational aids, as well as focus+context viewing, allowing enormous trees to fit within the fixed space of a computer screen or printed page. PMID- 11038341 TI - Intracellular mechanisms of antidepressant drug action. AB - The action of antidepressant drugs on monoamines such as norepinephrine and serotonin has been described for three decades. However, more-recent research has looked beyond cell surface receptors to transductional cascades and gene expression. Antidepressant drug therapies seem to share several mechanisms involved in either activating the adenylyl cyclase-protein kinase A cascade or inhibiting the phospholipase C-protein kinase C mechanisms. These effects, ultimately, combine to regulate the expression of target genes. Several specific genes are known to be activated or inhibited by antidepressant therapies. Steady state levels of mRNA for glucocorticoid and mineralocorticoid receptors, brain derived neurotrophic factor and its receptor trkB, and preproenkephalin are enhanced, whereas those for corticotropin-releasing hormone, c-fos,N-methyl-D aspartate receptor subunits, and nerve-growth factor 1A are reduced. New molecular genetic methods for identifying differentially expressed genes will aid in the development of targets for wholly new generations of antidepressant drug therapies. PMID- 11038342 TI - Predispositions, personality traits, and posttraumatic stress disorder. AB - This review suggests an explanation for the finding that trauma is a necessary but insufficient condition for the development of posttraumatic stress disorder. Predispositions affecting vulnerability to stress can help to account for discrepancies between traumatic exposure and pathological outcome. Such predispositions may be rooted in personality traits that shape the cognitive processing of stressful events, but they are also influenced by both prior life experiences and social supports. All of these observations are consistent with a biopsychosocial model of posttraumatic stress disorder. PMID- 11038343 TI - Philosophy of mind in the clinic: the relation between causal and meaningful explanation in psychiatry. AB - Conceptual dichotomies between mind and brain, psychology and neuroscience, meaning and causation, and fact and value confound thinking in philosophy of mind, clinical psychiatry, and psychiatric ethics. Paul Churchland's theory of eliminative materialism highlights these dichotomies, stating that advances in neuroscience have restricted, and eventually will eliminate, any need for psychology. The core principles of this theory are questionable, because psychiatrists still need psychology and perhaps always will. The main argument in this essay is that even in cases of well-defined brain pathology (where eliminative materialism seems most plausible), psychological concepts remain critical. Thus, philosophers and psychiatrists should generate conceptual models that lead not to eliminativism but to explanatory pluralism, which is the pragmatic integration of diverse concepts toward the end of better handling clinical challenges. The contributions of Karl Jaspers in opposition to eliminative materialism and in support of pluralism are discussed. Jaspers delineated the role of meaningful psychological explanation in a psychiatry rooted in neuroscientific explanation. However, his notion that meaningful states do not have causal power is disputable and has come under fire in recent philosophy of mind and cognitive neuroscience, which highlight the possibility that meaningful psychological states can be causally significant. This idea has implications for psychiatric ethics and the fact/value debate. PMID- 11038344 TI - The course of Tourette's disorder: a literature review. AB - With the goal of evaluating the available literature on the course of Tourette's disorder, we conducted a systematic literature search through electronic databases for pertinent scientific articles in English with a minimum of 20 subjects. We also examined bibliographies of papers identified in this manner for additional sources. We found only 16 articles; most consisted of retrospective reports on treated samples. Overall, the available literature suggests that Tourette's disorder follows a remitting course in a sizeable number of individuals. Little has been published regarding predictors of remission or persistence. More work is needed using longitudinal prospective studies to better define the course and outcome of Tourette's disorder. PMID- 11038345 TI - Overscrutinized and undersupported: a resident treats a "capitated" patient. PMID- 11038346 TI - Psychiatry and primary care: new directions in education. PMID- 11038347 TI - MEK inhibition enhances paclitaxel-induced tumor apoptosis. AB - The anti-cancer drug paclitaxel (Taxol) alters microtubule assembly and activates pro-apoptotic signaling pathways. Previously, we and others found that paclitaxel activates endogenous JNK in tumor cells, and the activation of JNK contributes to tumor cell apoptosis. Here we find that paclitaxel activates the prosurvival MEK/ERK pathway, which conversely may compromise the efficacy of paclitaxel. Hence, a combination treatment of paclitaxel and MEK inhibitors was pursued to determine whether this treatment could lead to enhanced apoptosis. The inhibition of MEK/ERK with a pharmacologic inhibitor, U0126, together with paclitaxel resulted in a dramatic enhancement of apoptosis that is four times more than the additive value of the two drugs alone. Enhanced apoptosis was verified by the terminal transferase-mediated dUTP nick end labeling assay, by an enzyme-linked immunosorbent assay for histone-associated DNA fragments, and by flow cytometric analysis for DNA content. Specificity of the pharmacologic inhibitor was confirmed by the use of (a) a second MEK/ERK inhibitor and (b) a transdominant negative MEK. Enhanced apoptosis was verified in breast, ovarian, and lung tumor cell lines, suggesting this effect is not cell type-specific. This is the first report of enhanced apoptosis detected in the presence of paclitaxel and MEK inhibition and suggests a new anticancer strategy. PMID- 11038348 TI - Sequence-specific DNA binding activity of RNA helicase A to the p16INK4a promoter. AB - p16(INK4a) is frequently altered in human cancer, often through epigenetically mediated transcriptional silencing. However, little is known about the transcriptional regulation of this gene. To learn more about such control, we initiated studies of proteins that bind to the promoter in cancer cells that do, and do not, express the gene. We identify RNA helicase A (RHA) as a protein that binds much better to the p16(INK4a) promoter in the expressing cells. RHA has not previously been characterized to manifest sequence-specific DNA interaction but does so to the sequence 5' CGG ACC GCG TGC GC 3' in the p16(INK4a) promoter. The Drosophila homologue to RHA, maleless (Mle), functions in the fly for 2-fold activation of male X-chromosome genes. In our experimental setting, RHA induces a similar modest up-regulation of the p16(INK4a) promoter that is dependent upon its sequence-specific interaction. Mle colocalizes with hyperacetylated H4Ac16 on the X-chromosome and some autosomal loci. The decreased binding of RHA to p16(INK4a) in our cells, where the gene is transcriptionally inactive, is associated with decreased amounts of RHA that immunoprecipitate with acetylated lysine antibodies. Finally, we show RHA to be a cellular substrate for caspase-3, which decreases its sequence-specific binding to p16(INK4a) by cleavage of the N terminus. Thus, we have identified a new protein interaction with the p16(INK4a) promoter that involves an important protein for transcriptional modulation. This interaction is decreased in cancer cells, where this gene is aberrantly transcriptionally silent. PMID- 11038349 TI - Structural events during the refolding of an all beta-sheet protein. AB - The refolding kinetics of the 140-residue, all beta-sheet, human fibroblast growth factor (hFGF-1) is studied using a variety of biophysical techniques such as stopped-flow fluorescence, stopped-flow circular dichroism, and quenched-flow hydrogen exchange in conjunction with multidimensional NMR spectroscopy. Urea induced unfolding of hFGF-1 under equilibrium conditions reveals that the protein folds via a two-state (native <--> unfolded) mechanism without the accumulation of stable intermediates. However, measurement of the unfolding and refolding rates in various concentrations of urea shows that the refolding of hFGF-1 proceeds through accumulation of kinetic intermediates. Results of the quenched flow hydrogen exchange experiments reveal that the hydrogen bonds linking the N- and C-terminal ends are the first to form during the refolding of hFGF-1. The basic beta-trefoil framework is provided by the simultaneous formation of beta strands I, IV, IX, and X. The other beta-strands comprising the beta-barrel structure of hFGF-1 are formed relatively slowly with time constants ranging from 4 to 13 s. PMID- 11038350 TI - Transcriptional regulation of the human acid alpha-glucosidase gene. Identification of a repressor element and its transcription factors Hes-1 and YY1. AB - Acid alpha-glucosidase, the product of a housekeeping gene, is a lysosomal enzyme that degrades glycogen. A deficiency of this enzyme is responsible for a recessively inherited myopathy and cardiomyopathy, glycogenesis type II. We have previously demonstrated that the human acid alpha-glucosidase gene expression is regulated by a silencer within intron 1, which is located in the 5'-untranslated region. In this study, we have used deletion analysis, electrophoretic mobility shift assay, and footprint analysis to further localize the silencer to a 25-base pair element. The repressive effect on the TK promoter was about 50% in both orientations in expression plasmid, and two transcriptional factors were identified with antibodies binding specifically to the element. Mutagenesis and functional analyses of the element demonstrated that the mammalian homologue 1 of Drosophila hairy and Enhancer of split (Hes-1) binding to an E box (CACGCG) and global transcription factor-YY1 binding to its core site function as a transcriptional repressor. Furthermore, the overexpression of Hes-1 significantly enhanced the repressive effect of the silencer element. The data should be helpful in understanding the expression and regulation of the human acid alpha glucosidase gene as well as other lysosomal enzyme genes. PMID- 11038351 TI - Rhodocytin induces platelet aggregation by interacting with glycoprotein Ia/IIa (GPIa/IIa, Integrin alpha 2beta 1). Involvement of GPIa/IIa-associated src and protein tyrosine phosphorylation. AB - Although glycoprotein Ia/IIa (GPIa/IIa, integrin alpha(2)beta(1)) has established its role as a collagen receptor, it remains unclear whether GPIa/IIa mediates activation signals. In this study, we show that rhodocytin, purified from the Calloselasma rhodostoma venom, induces platelet aggregation, which can be blocked by anti-GPIa monoclonal antibodies. Studies with rhodocytin-coupled beads and liposomes loaded with recombinant GPIa/IIa demonstrated that rhodocytin directly binds to GPIa/IIa independently of divalent cations. In vitro kinase assays and Western blotting of GPIa immunoprecipitates revealed that Src and Lyn constitutively associate with GPIa/IIa and that Src activity increases transiently after rhodocytin stimulation. Src specifically associates with p130 Crk-associated substrate (Cas) in a manner dependent upon Cas phosphorylation, suggesting that Src is responsible for Cas tyrosine phosphorylation. While all these phenomena occur early after rhodocytin stimulation in a cAMP-resistant manner, tyrosine phosphorylation of Syk and phospholipase Cgamma2, intracellular Ca(2+) mobilization, and platelet aggregation occur later in a cAMP-sensitive manner. Cytochalasin D, which interferes with actin polymerization and blocks receptor clustering, inhibits all the rhodocytin-mediated signals we examined in this study. We suggest that rhodocytin, by clustering GPIa/IIa, activates GPIa/IIa-associated Src, which then mediates downstream activation signals. PMID- 11038352 TI - Ligand-independent dimerization and activation of the oncogenic Xmrk receptor by two mutations in the extracellular domain. AB - Overexpression of the oncogenic receptor tyrosine kinase ONC-Xmrk is the first step in the development of hereditary malignant melanoma in the fish Xiphophorus. However, overexpression of its proto-oncogene counterpart (INV-Xmrk) is not sufficient for the oncogenic function of the receptor. Compared with INV-Xmrk, the ONC-Xmrk receptor displays 14 amino acid changes, suggesting the presence of activating mutations. To identify such activating mutations, a series of chimeric and mutant receptors were studied. None of the mutations present in the intracellular domain was found to be involved in receptor activation. In the extracellular domain, we found two mutations responsible for activation of the receptor. One is the substitution of a conserved cysteine (C578S) involved in intramolecular disulfide bonding. The other is a glycine to arginine exchange (G359R) in subdomain III. Either mutation leads to constitutive dimer formation and thereby to activation of the ONC-Xmrk receptor. Besides, the presence of these mutations slows down the processing of the Xmrk receptor in the endoplasmic reticulum, which is apparent as an incomplete glycosylation. PMID- 11038353 TI - Specific binding of nisin to the peptidoglycan precursor lipid II combines pore formation and inhibition of cell wall biosynthesis for potent antibiotic activity. AB - Unlike numerous pore-forming amphiphilic peptide antibiotics, the lantibiotic nisin is active in nanomolar concentrations, which results from its ability to use the lipid-bound cell wall precursor lipid II as a docking molecule for subsequent pore formation. Here we use genetically engineered nisin variants to identify the structural requirements for the interaction of the peptide with lipid II. Mutations affecting the conformation of the N-terminal part of nisin comprising rings A through C, e.g. [S3T]nisin, led to reduced binding and increased the peptide concentration necessary for pore formation. The binding constant for the S3T mutant was 0.043 x 10(7) m(-1) compared with 2 x 10(7) m(-1) for the wild-type peptide, and the minimum concentration for pore formation increased from the 1 nm to the 50 nm range. In contrast, peptides mutated in the flexible hinge region, e.g. [DeltaN20/DeltaM21]nisin, were completely inactive in the pore formation assay, but were reduced to some extent in their in vivo activity. We found the remaining in vivo activity to result from the unaltered capacity of the mutated peptide to bind to lipid II and thus to inhibit its incorporation into the peptidoglycan network. Therefore, through interaction with the membrane-bound cell wall precursor lipid II, nisin inhibits peptidoglycan synthesis and forms highly specific pores. The combination of two killing mechanisms in one molecule potentiates antibiotic activity and results in nanomolar MIC values, a strategy that may well be worth considering for the construction of novel antibiotics. PMID- 11038354 TI - The proteoglycans aggrecan and Versican form networks with fibulin-2 through their lectin domain binding. AB - Aggrecan, versican, neurocan, and brevican are important components of the extracellular matrix in various tissues. Their amino-terminal globular domains bind to hyaluronan, but the function of their carboxyl-terminal globular domains has long remained elusive. A picture is now emerging where the C-type lectin motif of this domain mediates binding to other extracellular matrix proteins. We here demonstrate that aggrecan, versican, and brevican lectin domains bind fibulin-2, whereas neurocan does not. As expected for a C-type lectin, the interactions are calcium-dependent, with K(D) values in the nanomolar range as measured by surface plasmon resonance. Solid phase competition assays with previously identified ligands demonstrated that fibulin-2 and tenascin-R bind the same site on the proteoglycan lectin domains. Fibulin-1 has affinity for the common site on versican but may bind to a different site on the aggrecan lectin domain. By using deletion mutants, the interaction sites for aggrecan and versican lectin domains were mapped to epidermal growth factor-like repeats in domain II of fibulin-2. Affinity chromatography and solid phase assays confirmed that also native full-length aggrecan and versican bind the lectin domain ligands. Electron microscopy confirmed the mapping and demonstrated that hyaluronan-aggrecan complexes can be cross-linked by the fibulins. PMID- 11038355 TI - Neuronal nitric-oxide synthase mutant (Ser-1412 --> Asp) demonstrates surprising connections between heme reduction, NO complex formation, and catalysis. AB - Rat neuronal NO synthase (nNOS) contains an Akt-dependent phosphorylation motif in its reductase domain. We mutated a target residue in that site (Ser-1412 to Asp) to mimic phosphorylation and then characterized the mutant using conventional and stopped-flow spectroscopies. Compared with wild-type, S1412D nNOS catalyzed faster cytochrome c and ferricyanide reduction but displayed slower steady-state NO synthesis with greater uncoupling of NADPH oxidation. Paradoxically, the mutant had faster heme reduction, faster heme-NO complex formation, and greater heme-NO complex accumulation at steady state. To understand how these behaviors related to flavin and heme reduction rates, we utilized three soybean calmodulins (CaMs) that supported a range of slower flavin and heme reduction rates in mutant and wild-type nNOS. Reductase activity and two catalytic parameters (speed and amount of heme-NO complex formation) related directly to the speed of flavin and heme reduction. In contrast, steady-state NO synthesis increased, reached a plateau, and then fell at the highest rate of heme reduction that was obtained with S1412D nNOS + CaM. Substituting with soybean CaM slowed heme reduction and increased steady-state NO synthesis by the mutant. We conclude the following. 1) The S1412D mutation speeds electron transfer out of the reductase domain. 2) Faster heme reduction speeds intrinsic NO synthesis but diminishes NO release in the steady state. 3) Heme reduction displays an optimum regarding NO release during steady state. The unique behavior of S1412D nNOS reveals the importance of heme reduction rate in controlling steady-state activity and suggests that nNOS already has a near-optimal rate of heme reduction. PMID- 11038356 TI - A kinetic simulation model that describes catalysis and regulation in nitric oxide synthase. AB - After initiating NO synthesis a majority of neuronal NO synthase (nNOS) quickly partitions into a ferrous heme-NO complex. This down-regulates activity and increases enzyme K(m,O(2)). To understand this process, we developed a 10-step kinetic model in which the ferric heme-NO enzyme forms as the immediate product of catalysis, and then partitions between NO dissociation versus reduction to a ferrous heme-NO complex. Rate constants used for the model were derived from recent literature or were determined here. Computer simulations of the model precisely described both pre-steady and steady-state features of nNOS catalysis, including NADPH consumption and NO production, buildup of a heme-NO complex, changes between pre-steady and steady-state rates, and the change in enzyme K(m,O(2)) in the presence or absence of NO synthesis. The model also correctly simulated the catalytic features of nNOS mutants W409F and W409Y, which are hyperactive and display less heme-NO complex formation in the steady state. Model simulations showed how the rate of heme reduction influences several features of nNOS catalysis, including populations of NO-bound versus NO-free enzyme in the steady state and the rate of NO synthesis. The simulation predicts that there is an optimum rate of heme reduction that is close to the measured rate in nNOS. Ratio between NADPH consumption and NO synthesis is also predicted to increase with faster heme reduction. Our kinetic model is an accurate and versatile tool for understanding catalytic behavior and will provide new perspectives on NOS regulation. PMID- 11038357 TI - Identification of two Entamoeba histolytica sequence-specific URE4 enhancer binding proteins with homology to the RNA-binding motif RRM. AB - To study transcriptional regulation in the lower branching eukaryote Entamoeba histolytica, we have identified two sequence-specific DNA-binding proteins that recognize the upstream regulatory element URE4, an enhancer that regulates expression of the Gal/GalNAc lectin heavy subunit gene hgl5. A chromatographic purification of E. histolytica nuclear extracts by gel filtration, cation exchange, and sequence-specific DNA affinity chromatography led to a 700-fold increase in URE4 binding activity and the appearance of two dominant protein species with molecular masses of 28 and 18 kDa. These proteins, termed E. histolytica enhancer-binding proteins 1 and 2 (EhEBP1 and EhEBP2), were sequenced by tandem mass spectroscopy and their corresponding cDNA clones identified. Recombinant EhEBP1 and EhEBP2 were able to bind double-stranded oligonucleotides bearing the URE4 motif in a sequence-specific manner, and antibodies raised against EhEBP1 were able to interfere with the formation of URE4-protein complexes in crude nuclear extracts. Overexpression of EhEBP1 in E. histolytica trophozoites resulted in a 7-fold drop in promoter activity in transiently transfected reporter gene constructs when the URE4 motif was present, confirming its ability to specifically recognize the URE4 motif and suggesting that additional cofactors may be required for transcriptional activation by URE4. Further characterization and identification of binding partners for EhEBP1 and EhEBP2, the first proteins with demonstrated sequence-specific DNA binding activity to be identified in E. histolytica, should provide new insights into transcriptional regulation in this protozoan parasite. PMID- 11038358 TI - Cystic fibrosis transmembrane conductance regulator Cl- channels with R domain deletions and translocations show phosphorylation-dependent and -independent activity. AB - Phosphorylation of the R domain regulates cystic fibrosis transmembrane conductance regulator Cl- channel activity. Earlier studies suggested that the R domain controls activity via more than one mechanism; a phosphorylated R domain may stimulate activity, and an unphosphorylated R domain may prevent constitutive activity, i.e. opening with ATP alone. However, the mechanisms responsible for these two regulatory properties are not understood. In this study we asked whether the two effects are dependent on its position in the protein and whether smaller regions from the R domain mediate the effects. We found that several portions of the R domain conferred phosphorylation-stimulated activity. This was true whether the R domain sequences were present in their normal location or were translocated to the C terminus. We also found that some parts of the R domain could be deleted without inducing constitutive activity. However, when residues 760-783 were deleted, channels opened without phosphorylation. Translocation of the R domain to the C terminus did not prevent constitutive activity. These results suggest that different parts of the phosphorylated R domain can stimulate activity and that their location within the protein is not critical. In contrast, prevention of constitutive activity required a short specific sequence that could not be moved to the C terminus. These results are consistent with a recent model of an R domain composed primarily of random coil in which more than one phosphorylation site is capable of stimulating channel activity, and net activity reflects interactions between multiple sites in the R domain and the rest of the channel. PMID- 11038359 TI - Transcription factors Zic1 and Zic2 bind and transactivate the apolipoprotein E gene promoter. AB - We have used the yeast one-hybrid system to identify transcription factors that bind to specific sequences in proximal regions of the apolipoprotein E gene promoter. The sequence between -163 and -124, that has been previously defined as a functional promoter element, was used as a bait to screen a human brain cDNA library. Ten cDNA clones that encoded portions of the human Zic1 (five clones) and Zic2 (five clones) transcription factors were isolated. Electrophoretic mobility shift assays confirmed the presence of a binding site for Zic1 and Zic2 in the -136/-125 region. Displacement of binding with oligonucleotides derived from adjacent sequences within the APOE promoter revealed the existence of two additional Zic-binding sequences in this promoter. These sequences were identified by electrophoretic mobility shift assays and mutational analysis in regions -65/-54 and -185/-174. Cotransfection of Zic1 and Zic2 expression vector and different APOE promoter-luciferase reporter constructs in U87 glioblastoma cell line showed that the three binding sites partially contributed to the trans stimulation of the luciferase reporter. Ectopic expression of Zic1 and Zic2 in U87 cells also trans-stimulated the expression of the endogenous gene, increasing the amount of apolipoprotein E produced by glial cells. These data indicate that Zic proteins might contribute to the transcriptional activity of the apolipoprotein E gene and suggest that apolipoprotein E could mediate some of the developmental processes in which Zic proteins are involved. PMID- 11038360 TI - Functional dissection of the LysR-type CysB transcriptional regulator. Regions important for DNA binding, inducer response, oligomerization, and positive control. AB - CysB is a tetrameric LysR-type transcriptional regulator that acts as an activator of cys regulon genes and as an autorepressor. Positive control of cys genes requires the presence of the inducer N-acetylserine. Following random and site-directed mutagenesis of the cysB gene, 20 CysB variants were isolated. Six single amino acid substitutions within the N terminus of CysB abolished the DNA binding ability of the protein. Seven mutations in the central region of CysB affected its response to the inducer. Four of these CysB mutants retained repressing activity, but lost their activating function in vivo. Their DNA binding characteristics were consistent with an inability to respond to acetylserine by a qualitative change in the DNA-protein interaction. Three of the single residue substitutions resulted in constitutive activity of CysB. The electrophoretic mobility of the complex formed by one of the CysBc variants with the cysP promoter suggested a dimeric state of this protein. Characteristics of six truncated CysB variants lacking 5-30 C-terminal residues indicated the involvement of the C terminus in the DNA binding, oligomerization, and stability of CysB. The single substitution Y27G resulted in the CysBpc variant, able to bind DNA and to respond to the inducer by a qualitative change in the DNA-protein complex, but defective in the positive control of the cysP promoter. PMID- 11038361 TI - High resolution structure of the phosphohistidine-activated form of Escherichia coli cofactor-dependent phosphoglycerate mutase. AB - The active conformation of the dimeric cofactor-dependent phosphoglycerate mutase (dPGM) from Escherichia coli has been elucidated by crystallographic methods to a resolution of 1.25 A (R-factor 0.121; R-free 0.168). The active site residue His(10), central in the catalytic mechanism of dPGM, is present as a phosphohistidine with occupancy of 0.28. The structural changes on histidine phosphorylation highlight various features that are significant in the catalytic mechanism. The C-terminal 10-residue tail, which is not observed in previous dPGM structures, is well ordered and interacts with residues implicated in substrate binding; the displacement of a loop adjacent to the active histidine brings previously overlooked residues into positions where they may directly influence catalysis. E. coli dPGM, like the mammalian dPGMs, is a dimer, whereas previous structural work has concentrated on monomeric and tetrameric yeast forms. We can now analyze the sequence differences that cause this variation of quaternary structure. PMID- 11038362 TI - Impact of the reduced folate carrier on the accumulation of active thiamin metabolites in murine leukemia cells. AB - The thiamin transporter encoded by SLC19A2 and the reduced folate carrier (RFC1) share 40% homology at the protein level, but the thiamin transporter does not mediate transport of folates. By using murine leukemia cell lines that express no, normal, or high levels of RFC1, we demonstrate that RFC1 does not mediate thiamin influx. However, high level RFC1 expression substantially reduced accumulation of the active thiamin coenzyme, thiamin pyrophosphate (TPP). This decreased level of TPP, synthesized intracellularly from imported thiamin, resulted from RFC1-mediated efflux of TPP. This conclusion was supported by the following observations. (i) Efflux of intracellular TPP was increased in cells with high expression of RFC1. (ii) Methotrexate inhibits TPP influx. (iii) TPP competitively inhibits methotrexate influx. (iv) Loading cells, which overexpress RFC1 to high levels of methotrexate to inhibit competitively RFC1-mediated TPP efflux, augment TPP accumulation. (v) There was an inverse correlation between thiamin accumulation and RFC1 activity in cells grown at a physiological concentration of thiamin. The modulation of thiamin accumulation by RFC1 in murine leukemia cells suggests that this carrier may play a role in thiamin homeostasis and could serve as a modifying factor in thiamin nutritional deficiency as well as when the high affinity thiamin transporter is mutated. PMID- 11038363 TI - Impaired prohormone convertases in Cpe(fat)/Cpe(fat) mice. AB - A spontaneous point mutation in the coding region of the carboxypeptidase E (CPE) gene results in a loss of CPE activity that correlates with the development of late onset obesity (Nagert, J. K., Fricker, L. D., Varlamov, O., Nishina, P. M., Rouille, Y., Steiner, D. F., Carroll, R. J., Paigen, B. J., and Leiter, E. H. (1995) Nat. Genet. 10, 135-142). Examination of the level of neuropeptides in these mice showed a decrease in mature bioactive peptides as a result of a decrease in both carboxypeptidase and prohormone convertase activities. A defect in CPE is not expected to affect endoproteolytic processing. In this report we have addressed the mechanism of this unexpected finding by directly examining the expression of the major precursor processing endoproteases, prohormone convertases PC1 and PC2 in Cpe(fat) mice. We found that the levels of PC1 and PC2 are differentially altered in a number of brain regions and in the pituitary. Since these enzymes have been implicated in the generation of neuroendocrine peptides (dynorphin A-17, beta-endorphin, and alpha- melanocyte-stimulating hormone) involved in the control of feeding behavior and body weight, we compared the levels of these peptides in Cpe(fat) and wild type animals. We found a marked increase in the level of dynorphin A-17, a decrease in the level of alpha melanocyte-stimulating hormone, and an alteration in the level of C-terminally processed beta-endorphin. These results suggest that the impairment in the level of these and other peptides involved in body weight regulation is mainly due to an alteration in carboxypeptidase and prohormone convertase activities and that this may lead to the development of obesity in these animals. PMID- 11038364 TI - Dissection of a nuclear localization signal. AB - The regulated process of protein import into the nucleus of a eukaryotic cell is mediated by specific nuclear localization signals (NLSs) that are recognized by protein import receptors. This study seeks to decipher the energetic details of NLS recognition by the receptor importin alpha through quantitative analysis of variant NLSs. The relative importance of each residue in two monopartite NLS sequences was determined using an alanine scanning approach. These measurements yield an energetic definition of a monopartite NLS sequence where a required lysine residue is followed by two other basic residues in the sequence K(K/R)X(K/R). In addition, the energetic contributions of the second basic cluster in a bipartite NLS ( approximately 3 kcal/mol) as well as the energy of inhibition of the importin alpha importin beta-binding domain ( approximately 3 kcal/mol) were also measured. These data allow the generation of an energetic scale of nuclear localization sequences based on a peptide's affinity for the importin alpha-importin beta complex. On this scale, a functional NLS has a binding constant of approximately 10 nm, whereas a nonfunctional NLS has a 100 fold weaker affinity of 1 microm. Further correlation between the current in vitro data and in vivo function will provide the foundation for a comprehensive quantitative model of protein import. PMID- 11038365 TI - A novel casein kinase 2 alpha-subunit regulates membrane protein traffic in the human hepatoma cell line HuH-7. AB - A previously isolated endocytic trafficking mutant (TRF1) isolated from HuH-7 cells is defective in the distribution of subpopulations of cell-surface receptors for asialoorosomucoid (asialoglycoprotein receptor (ASGR)), transferrin, and mannose-terminating glycoproteins. The pleiotropic phenotype of TRF1 also includes an increased sensitivity to Pseudomonas toxin and deficient assembly and function of gap junctions. HuH-7xTRF1 hybrids exhibited a normal subcellular distribution of ASGR, consistent with the TRF1 mutation being recessive. A cDNA expression library derived from HuH-7 mRNA was transfected into TRF1 cells, which were subsequently selected for resistance to Pseudomonas toxin. Sequence analysis of a recovered cDNA revealed a unique isoform of casein kinase 2 (CK2), CK2alpha". Western blot analysis of TRF1 proteins revealed a 60% reduction in total CK2alpha expression. Consistent with this finding, the hybrids HuH-7xHuH-7 and HuH-7xTRF1 expressed equivalent amounts of total CK2alpha. Immunoblots using antibodies against peptides unique to the previously described CK2 isoforms CK2alpha and CK2alpha' and the novel CK2alpha" isoform showed that, although TRF1 and parental HuH-7 cells expressed comparable amounts of CK2alpha and CK2alpha', the mutant did not express CK2alpha". Based on the genomic DNA sequence, RNA transcripts encoding CK2alpha" apparently originate from alternative splicing of a primary transcript. Protein overexpression following transfection of TRF1 cells with cDNAs encoding either CK2alpha or the newly cloned CK2alpha" restored the parental HuH-7 phenotype, including Pseudomonas toxin resistance, cell-surface ASGR binding activity, phosphorylation, and the assembly of gap junctions. This study suggests that HuH-7 cells express at least three CK2alpha isoforms and that the pleiotropic TRF1 phenotype is a consequence of a reduction in total CK2 expression. PMID- 11038366 TI - Carboxymethylation of the PP2A catalytic subunit in Saccharomyces cerevisiae is required for efficient interaction with the B-type subunits Cdc55p and Rts1p. AB - Protein phosphatase 2A (PP2A) is an essential eukaryotic serine/threonine phosphatase known to play important roles in cell cycle regulation. Association of different B-type targeting subunits with the heterodimeric core (A/C) enzyme is known to be an important mechanism of regulating PP2A activity, substrate specificity, and localization. However, how the binding of these targeting subunits to the A/C heterodimer might be regulated is unknown. We have used the budding yeast Saccharomyces cerevisiae as a model system to investigate the hypothesis that covalent modification of the C subunit (Pph21p/Pph22p) carboxyl terminus modulates PP2A complex formation. Two approaches were taken. First, S. cerevisiae cells were generated whose survival depended on the expression of different carboxyl-terminal Pph21p mutants. Second, the major S. cerevisiae methyltransferase (Ppm1p) that catalyzes the methylation of the PP2A C subunit carboxyl-terminal leucine was identified, and cells deleted for this methyltransferase were utilized for our studies. Our results demonstrate that binding of the yeast B subunit, Cdc55p, to Pph21p was disrupted by either acidic substitution of potential carboxyl-terminal phosphorylation sites on Pph21p or by deletion of the gene for Ppm1p. Loss of Cdc55p association was accompanied in each case by a large reduction in binding of the yeast A subunit, Tpd3p, to Pph21p. Moreover, decreased Cdc55p and Tpd3p binding invariably resulted in nocodazole sensitivity, a known phenotype of CDC55 or TPD3 deletion. Furthermore, loss of methylation also greatly reduced the association of another yeast B-type subunit, Rts1p. Thus, methylation of Pph21p is important for formation of PP2A trimeric and dimeric complexes, and consequently, for PP2A function. Taken together, our results indicate that methylation and phosphorylation may be mechanisms by which the cell dynamically regulates PP2A complex formation and function. PMID- 11038367 TI - Chloroplast ribonucleoproteins function as a stabilizing factor of ribosome-free mRNAs in the stroma. AB - Post-transcriptional RNA processing is an important step in the regulation of chloroplast gene expression, and a number of chloroplast ribonucleoproteins (cpRNPs) are likely to be involved in this process. The major tobacco cpRNPs are composed of five species: cp28, cp29A, cp29B, cp31, and cp33 and these are divided into three groups (I, II, and III). By immunoprecipitation, gel filtration, and Western blot analysis, we demonstrated that these cpRNPs are abundant stromal proteins that exist as complexes with ribosome-free mRNAs. Many ribosome-free psbA mRNAs coprecipitate with cpRNPs, indicating that the majority of stromal psbA mRNAs are associated with cpRNPs. In addition, an in vitro mRNA degradation assay indicated that exogenous psbA mRNA is more rapidly degraded in cpRNP-depleted extracts than in nondepleted extracts. When the depleted extract was reconstituted with recombinant cpRNPs, the psbA mRNA in the extract was protected from degradation to a similar extent as the psbA mRNA in the nondepleted extract. Moreover, restoration of the stabilizing activity varied following addition of individual group-specific cpRNPs alone or in combination. When the five cpRNPs were supplemented in the depleted extract, full activity was restored. We propose that these cpRNPs act as stabilizing factors for nonribosome bound mRNAs in the stroma. PMID- 11038368 TI - GATA-4 and serum response factor regulate transcription of the muscle-specific carnitine palmitoyltransferase I beta in rat heart. AB - Transcriptional regulation of nuclear encoded mitochondrial proteins is dependent on nuclear transcription factors that act on genes encoding key components of mitochondrial transcription, replication, and heme biosynthetic machinery. Cellular factors that target expression of proteins to the heart have been well characterized with respect to excitation-contraction coupling. No information currently exists that examines whether parallel transcriptional mechanisms regulate nuclear encoded expression of heart-specific mitochondrial isoforms. The muscle CPT-Ibeta isoform in heart is a TATA-less gene that uses Sp-1 proteins to support basal expression. The rat cardiac fatty acid response element (-301/ 289), previously characterized in the human gene, is responsive to oleic acid following serum deprivation. Deletion and mutational analysis of the 5'-flanking sequence of the carnitine palmitoyltransferase Ibeta (CPT-Ibeta) gene defines regulatory regions in the -391/+80 promoter luciferase construct. When deleted or mutated constructs were individually transfected into cardiac myocytes, CPT I/luciferase reporter gene expression was significantly depressed at sites involving a putative MEF2 sequence downstream from the fatty acid response element and a cluster of heart-specific regulatory regions flanked by two Sp1 elements. Each site demonstrated binding to cardiac nuclear proteins and competition specificity (or supershifts) with oligonucleotides and antibodies. Individual expression vectors for Nkx2.5, serum response factor (SRF), and GATA4 enhanced CPT-I reporter gene expression 4-36-fold in CV-1 cells. Although cotransfection of Nkx and SRF produced additive luciferase expression, the combination of SRF and GATA-4 cotransfection resulted in synergistic activation of CPT-Ibeta. The results demonstrate that SRF and the tissue-restricted isoform, GATA-4, drive robust gene transcription of a mitochondrial protein highly expressed in heart. PMID- 11038369 TI - Update on adenovirus and its vectors. PMID- 11038370 TI - Blood clearance rates of adenovirus type 5 in mice. AB - Persistence of adenovirus type 5 in blood has implications for the pathogenicity of the virus infection and for the use of this virus in oncolysis and gene therapy. In this study, the kinetics of adenovirus clearance from blood in mice has been evaluated. After a single inoculation of concentrated virus into the vena cava, virus half-life was less than 2 min. Depletion of Kupffer cells (KC) resulted in increased viraemia. After tail-vein injection, virus and latex beads co-localized within KC. An important factor in clearance by KC is the negative charge of particles. Deletion of the hexon hypervariable region 1 acidic stretch decreased the negative charge of the virion but it did not increase blood persistence. Coating with PEG ('PEGylation') reduced the clearance rate but also reduced infectivity. PMID- 11038371 TI - Pyrogenicity of human adenoviruses. AB - High doses (>1.56x10(7) p.f.u.) of purified preparations of human adenovirus types 3, 5 and 8 exhibited definite pyrogenic activity when injected intravenously into rabbits. Complete pyrogenic tolerance was obtained not only with homologous types but also with heterologous types of adenovirus. No pyrogenic cross-tolerance was observed between each of these three adenovirus types and paramyxovirus pyrogen or bacterial lipopolysaccharide. Adenovirus pyrogenicity was retained after UV-inactivation, whereas it was inactivated by heating at 56 degrees C for 30 min. Adenovirus pyrogenicity was not neutralized by mixing with homologous type-specific antiserum but non-pyrogenic doses (10(7) p.f.u.) of adenovirus types 3, 5 and 8 became highly pyrogenic in the presence of type-specific antibodies at the optimal virus:antibody ratio. This enhanced pyrogenicity depended upon the virus-antibody complex. From these results, it is probable that the pyrogenic activity of the virus-antibody complex, rather than the pyrogenic activity of the virions, is the main contributor to fever in adenovirus infection under actual physiological conditions. PMID- 11038372 TI - Interaction with CBP/p300 enables the bovine papillomavirus type 1 E6 oncoprotein to downregulate CBP/p300-mediated transactivation by p53. AB - The E6 oncoprotein of bovine papillomavirus type 1 (BPV-1) can transform cells independently of p53 degradation. The precise mechanisms underlying this transformation are not yet completely understood. Here it is shown that BPV-1 E6 interacts with CBP/p300 in the same way as described for the E6 proteins of oncogenic human papillomaviruses. This interaction results in an inhibition of the transcriptional coactivator function of CBP/p300 required by p53 and probably by other transcription factors. The comparison of the CBP/p300-binding properties of BPV-1 E6 mutants previously characterized in transcription and transformation studies suggests (i) that the E6-CBP/p300 interaction may be necessary, but not sufficient, for cell transformation, and (ii) that the transcriptional activator function, inherent to the E6 protein, is not derived from forming a complex with CBP/p300. PMID- 11038373 TI - VP1 DNA sequences of JC and BK viruses detected in urine of systemic lupus erythematosus patients reveal no differences from strains expressed in normal individuals. AB - The ubiquitous human polyomaviruses BK (BKV) and JC (JCV) persist with no adverse effects in immunocompetent individuals. Virus-induced pathogenesis has been linked to virus reactivation during impaired immune conditions. Previous studies have shown a significant difference between the VP1 DNA sequences of JCV obtained from control urine samples and those in progressive multifocal leukoencephalopathy brain samples. This difference could not be detected when comparing normal control urinary JCV DNA with DNA sequences from chronic progressive multiple sclerosis patients. Since BKV and JCV are readily activated in systemic lupus erythematosus (SLE) patients, the presence of specific strains, related to VP1 DNA sequences, was investigated in these patients. VP1 DNA sequences in 100 urine samples from 21 SLE patients and 75 urine samples from 75 healthy pregnant women were analysed and compared to previously reported sequences. The results show that the VP1 sequence profiles of JCV and BKV excreted by SLE patients do not differ significantly from those excreted by immunocompetent individuals. The European JCV subtypes 1A or 1B were represented among all JCV-positive urine specimens, while BKV VP1 sequences showed complete, or almost complete, identity with the MM or JL strains. Different urine samples from the same patient collected over a 1 year period were predominantly stable. BKV VP1 DNA in urine specimens from healthy pregnant women was only detected during the third trimester of their pregnancy. These results argue against SLE specific JCV and BKV strains and suggest reactivation of the viruses rather than recurrent re-infections of patients with SLE. PMID- 11038374 TI - Murine gammaherpesvirus-68 infection of and persistence in the central nervous system. AB - Murine gammaherpesvirus-68 (MHV-68) was originally isolated from a bank vole by passage through mouse brain. Given its ability to replicate in mouse brain and its subsequent reisolation from trigeminal ganglia, it was originally considered to be an alphaherpesvirus. Molecular studies have now firmly established MHV-68 to be a gammaherpesvirus. Other gammaherpesviruses have been suggested to cause and in some cases shown to cause neurological disease. Given the isolation history of MHV-68, we have studied the ability of this virus to gain access to, to replicate in and to persist in the mouse CNS. Following intranasal inoculation the virus was not generally neuroinvasive. However, in mice with a deletion of the type-I interferon receptor gene, peripheral virus titres are higher and perivascular CNS infection was observed. There was no evidence of virus spread via olfactory routes. Direct intracerebral inoculation of virus was fatal with widespread infection and destruction predominantly of meningeal and ependymal cells. Hippocampal pyramidal neurons, oligodendrocytes, Bergmann glia cells in the cerebellar cortex and neural progenitor cells in the rostral migratory stream were also infected. A similar infection was observed in younger mice. CNS infection following virus reactivation was investigated by implantation of infected glial cells. Implantation into a brain ventricle led to widespread fatal infection, principally involving ependymal and meningeal cells. Implantation into the striatum resulted in a predominantly neuronal infection. Implantation of cells into mice transiently treated with the antiviral thionucleoside analogue 2' deoxy-5-ethyl-beta-4'-thiouridine resulted in survival with detection of virus infected cells in the brain 1 year later. PMID- 11038375 TI - Latency-associated nuclear antigen of Kaposi's sarcoma-associated herpesvirus (human herpesvirus-8) binds ATF4/CREB2 and inhibits its transcriptional activation activity. AB - Latency-associated nuclear antigen (LANA), encoded by ORF 73 of Kaposi's sarcoma associated herpesvirus (KSHV; human herpesvirus-8), may play an important role in the persistence of the viral episome by tethering it to host chromosomes during mitosis. It also has been suggested from its amino acid sequence features that LANA may have transcription-regulatory activity. Here, it is reported that LANA interacts with activating transcription factor (ATF) 4/cAMP response element binding protein (CREB) 2, a member of the ATF/CREB family of transcription factors, and represses the transcriptional activation activity of ATF4/CREB2. Repression by LANA is independent of the DNA-binding ability of ATF4/CREB2, since LANA also represses transactivation of ATF4/CREB2 fused to the GAL4 DNA-binding domain and does not affect the DNA-binding ability of ATF4/CREB2 in an electrophoretic mobility shift assay. The putative leucine zipper domain of LANA is required for binding to the relatively conserved basic region/leucine zipper domain (bZIP) of ATF4/CREB2, suggesting that the interaction may involve leucine zipper dimerization. PMID- 11038376 TI - Characterization of the herpesvirus saimiri ORF73 gene product. AB - The herpesvirus saimiri (HVS) gene product encoded by ORF73 shares a limited homology with the ORF73 encoded protein of Kaposi's sarcoma-associated herpesvirus (KSHV). It has recently been shown that the KSHV ORF73 protein is expressed during a latent infection and co-localizes with host cell chromosomes, suggesting that it plays a role in episomal maintenance by tethering viral genomes to host cell chromosomes. At present the role of the HVS ORF73 gene product is unknown. However, the expression of HVS ORF73 in a stably transduced human carcinoma cell line, where the HVS genome persists as a non-integrated circular episome, has recently been shown. In this report, the characterization of the HVS ORF73 protein and the mapping of its functional domains are described. The results suggest that the HVS ORF73 gene encodes a 64 kDa nuclear protein. Moreover, the amino terminus contains two functional nuclear localization signals, whereas the carboxy terminus is required for the distinctive speckled nuclear distribution pattern as observed with both the HVS and KSHV ORF73 proteins. PMID- 11038377 TI - The antigenic domain 1 of human cytomegalovirus glycoprotein B contains an intramolecular disulphide bond. AB - Glycoprotein B (gB, gpUL55) is the major antigen recognized by the neutralizing humoral immune response against human cytomegalovirus (HCMV). The immunodominant region on gB is the antigenic domain 1 (AD-1), a complex structure that requires a minimal continuous sequence of more than 75 amino acids (aa 552-635) for antibody binding. In this study, the structural requirements for antibody binding to AD-1 have been determined. The domain was expressed in prokaryotic and eukaryotic systems and analysed in immunoblots under reducing and non-reducing conditions. In addition, AD-1 was purified in an immunologically active form and the concentration of sulphydryl groups was determined. The data clearly show that the only form that is recognized by antibodies is a disulphide-linked monomer of AD-1. The disulphide bond is formed between cysteines at amino acid positions 573 and 610 of gB. PMID- 11038378 TI - The in vivo effects of recombinant bovine herpesvirus-1 expressing bovine interferon-gamma. AB - To study the biological relevance of using bovine herpesvirus-1 (BHV-1) as a vector for expressing cytokines, a BHV-1 virus that expressed bovine interferon gamma (IFN-gamma) was constructed. This recombinant virus (BHV-1/IFNgamma) was then used to infect the natural host in a respiratory disease model. In vitro characterization of the recombinant interferon-gamma confirmed that the cytokine expressed in BHV-1-infected cells was biologically active. The in vivo effects of the recombinant IFN-gamma were then analysed during a primary infection and after reactivation of a latent infection. During the primary infection, similar body temperature, clinical responses and virus shedding were observed for calves infected with either recombinant BHV-1/IFNgamma or parental gC(-)/LacZ(+) virus. An analysis of cellular and humoral responses did not reveal any significant immunomodulation by BHV-1/IFNgamma during the primary infection. The stability and activity of recombinant IFN-gamma was also analysed following the establishment of a latent infection. The presence of recombinant IFN-gamma did not significantly alter virus shedding following reactivation. The isolation of reactivated BHV-1/IFNgamma virus confirmed that a functional IFN-gamma gene was retained during latency. Thus, herpesviruses may provide virus vectors that retain functional genes during latency and recrudescence. PMID- 11038379 TI - Vesicular stomatitis virus and pseudorabies virus induce a vig1/cig5 homologue in mouse dendritic cells via different pathways. AB - The homologous genes vig1 and cig5 were identified by differential display PCR as virus-induced genes in rainbow trout and humans, respectively. These genes are significantly related to sequences required for the biosynthesis of metal cofactors, but their function remains unknown. In this study, it is shown that the mouse homologue of vig1/cig5 was induced by vesicular stomatitis virus (VSV) and pseudorabies virus (PrV) in mouse spleen cells. Among a collection of cell lines from dendritic, myeloid, lymphoid or fibroblast lineages, only the dendritic cell line, D2SC1, showed expression of mvig after virus infection. This dendritic restriction was confirmed by our finding that mvig was also induced by both VSV and PrV in CD11c(++) spleen cells, separated by magnetic purification or derived from bone marrow precursor cells. Similar to the fish rhabdovirus viral haemorrhagic septicaemia virus in trout cells, VSV directly induced mvig in the dendritic cell line D2SC1, but the PrV-mediated induction required the integrity of the interferon pathway. This result indicates that mvig is interferon inducible like its fish and human homologues. Furthermore, mvig was also induced by LPS in bone marrow-derived cells. Thus, mvig expression seems to correlate with an activated state of dendritic cells subjected to different pathogen associated stimuli. PMID- 11038380 TI - Resistance to Rift Valley fever virus in Rattus norvegicus: genetic variability within certain 'inbred' strains. AB - Rift Valley fever virus (RVFV) is the causative agent of Rift Valley fever, a widespread disease of domestic animals and humans in sub-Saharan Africa. Laboratory rats have frequently been used as an animal model for studying the pathogenesis of Rift Valley fever. It is shown here that Lewis rats (LEW/mol) are susceptible to infection with RVFV, whereas Wistar-Furth (WF/mol) rats are resistant to RVFV infection. LEW/mol rats developed acute hepatitis and died after infection with RVFV strain ZH548, whereas WF/mol rats survived the infection. Cross-breeding of resistant WF/mol rats with susceptible LEW/mol rats demonstrated that resistance is segregated as a single dominant gene. Primary hepatocytes but not glial cells from WF/mol rats showed the resistant phenotype in cell culture, indicating that resistance was cell type-specific. Moreover, when cultured hepatocytes were stimulated with interferon (IFN) type I there was no indication of a regulatory role of IFN in the RVFV-resistance gene expression in WF/mol rats. Interestingly, previous reports have shown that LEW rats from a different breeding stock (LEW/mai) are resistant to RVFV infections, whereas WF/mai rats are susceptible. Thus, inbred rat strains seem to differ in virus susceptibility depending on their breeding histories. A better genetic characterization of inbred rat strains and a revision in nomenclature is needed to improve animal experimentation in the future. PMID- 11038381 TI - Heterologous protection against lethal A/HongKong/156/97 (H5N1) influenza virus infection in C57BL/6 mice. AB - The continual threat posed by newly emerging influenza virus strains is demonstrated by the recent outbreak of H5N1 influenza virus in Hong Kong. Currently, immunization against influenza virus infection is fairly adequate, but it is imperative that improved vaccines are developed that can protect against a variety of strains and be generated rapidly. Since humoral immunity is ineffective against serologically distinct viruses, one strategy would be to develop vaccines that emphasize cellular immunity. Here we report the successful protection of C57BL/6 mice from a lethal A/HK/156/97 (HK156) infection by immunizing first with an H9N2 isolate, A/Quail/HK/G1/97 (QHKG1), that harbours internal genes 98% homologous to HK156. This strategy also protected mice that are deficient in antibody production, indicating that the immunity is T-cell mediated. In the course of these studies, we generated a highly pathogenic H5N1 reassortant which implicated NP and PB2 as having an important contribution to pathogenesis when present with a highly cleavable H5. These results provide the first demonstration that protective cell-mediated immunity can be established against the highly virulent HK156 virus and have important implications for the development of novel strategies for the prevention and treatment of HK156 infection and the design of future influenza vaccines. PMID- 11038382 TI - Entry of influenza viruses into cells is inhibited by a highly specific protein kinase C inhibitor. AB - Following binding to cell surface sialic acid, entry of influenza viruses into cells is mediated by endocytosis. Productive entry of influenza virus requires the low-pH environment of the late endosome for fusion and release of the virus into the cytoplasm and transport of the virus genome into the nucleus. We investigated novel mechanisms to inhibit influenza virus infection using highly specific inhibitors of protein kinase C. We found that one inhibitor, bisindolylmaleimide I, prevented replication of influenza A virus in a dose dependent manner when added at the time of infection, but had little specific effect when added 2 h after infection had commenced. Virus yields dropped by more than 3 log units in the presence of micromolar levels of bisindolylmaleimide I. Influenza B virus replication was also inhibited by bisindolylmaleimide at micromolar concentrations. We carried out experiments to determine the point in infection that was blocked by bisindolylmaleimide I, and determined that entry of viral ribonucleoproteins (vRNPs) into the nucleus was prevented. Upon drug washout vRNP nuclear entry resumed, showing that bisindolylmaleimide I is reversible. Bisindolylmaleimide I did not affect virus binding and was apparently not acting as a weak base, because its effects were independent of the pH of the external growth medium. These experiments show that bisindolylmaleimide I blocks replication of different types of influenza virus in a dose-dependent and reversible manner, and that virus entry into the cell is inhibited. PMID- 11038383 TI - Role of CD4(+) and CD8(+) T cells in the prevention of measles virus-induced encephalitis in mice. AB - Depending on their major histocompatibility complex (MHC) haplotype, inbred mouse strains are either resistant (H2-d, BALB/c), susceptible (H2-k, C3H) or partially resistant (H2-dxk, BaCF1) to intracerebral infection with the neurotropic rodent adapted measles virus (MV) strain CAM/RBH. Here, mortality is demonstrated to be correlated directly with virus spread and virus replication in the CNS and to be inversely correlated with the activation of MV-specific T cells. Previously, it has been shown that primary CD4(+) T cells alone are protective in the resistant background. In the susceptible background, CD4(+) T cells acquire protective capacity after immunization with a newly defined CD4(+) T cell epitope peptide. In the partially resistant mice, CD4(+) T cells provide help for CD8(+) T cells and protect in cooperation with them. It seems that the lytic capacity of CD8(+) T cells is crucial in providing protection, as MV-specific L(d)-restricted CD8(+) T cells, which are highly lytic in vitro after transfer, protect naive animals against MV-induced encephalitis (MVE). In contrast, K(k)-restricted CD8(+) T cells with low lytic capacity do not protect. In the MVE model, CD4(+) T cells are able to protect either alone (resistant mice), through cooperation with CD8(+) T cells (intermediate susceptible) or after immunization as secondary T cells (susceptible mice). CD8(+) T cells are able to protect alone after immunization if they are cytolytic. Thus, susceptibility and resistance depend upon the functional composition of CD4(+) and CD8(+) T cells governed by the MHC haplotype. PMID- 11038384 TI - Binding of human respiratory syncytial virus to cells: implication of sulfated cell surface proteoglycans. AB - Binding of human respiratory syncytial virus (HRSV) to cultured cells was measured by flow cytometry. Using this assay and influenza virus as a control virus with a well-characterized receptor, a systematic search of cell surface molecules that might be implicated in HRSV binding was carried out. Treatment of cells with different enzymes or with other reagents suggested that heparin-like glycosaminoglycans (GAGs) were involved in attachment of HRSV, but not influenza virus, to host cells. This was further confirmed by a lack of binding of HRSV to CHO-K1 mutant cell lines deficient in glycosylation or GAGs biosynthesis and by an inhibition of binding after preincubation of virus with heparin and other GAGs. The degree of sulfation, more than the polysaccharide backbone of GAGs, seems to be critical for virus binding. PMID- 11038385 TI - Nucleotide sequences of the F, L and G protein genes of two non-A/non-B avian pneumoviruses (APV) reveal a novel APV subgroup. AB - Sequence analysis was performed of all or part of the genes encoding the fusion (F), polymerase (L) and attachment (G) proteins of two French non-A/non-B avian pneumovirus (APV) isolates (Fr/85/1 and Fr/85/2). The two isolates shared at least 99.7% nt and 99.0% aa sequence identity. Comparison with the F genes from subgroup A, subgroup B or Colorado APVs revealed nt and aa identities of 70.0-80. 5% and 77.6-97.2%, respectively, with the L gene sharing 76.1% nt and 85.3% aa identity with that of a subgroup A isolate. The Fr/85/1 and Fr/85/2 G genes comprised 1185 nt, encoding a protein of 389 aa. Common features with subgroup A and subgroup B G proteins included an amino-terminal membrane anchor, a high serine and threonine content, conservation of cysteine residues and a single extracellular region of highly conserved sequence proposed to be the functional domain involved in virus attachment to cellular receptors. However, the Fr/85/1 and Fr/85/2 G sequences shared at best 56.6% nt and 31.2% aa identity with subgroup A and B APVs, whereas these isolates share 38% aa identity. Phylogenetic analysis of the F, G and L genes of pneumoviruses suggested that isolates Fr/85/1 and Fr/85/2 belong to a previously unrecognized APV subgroup, tentatively named D. G-based oligonucleotide primers were defined for the specific molecular identification of subgroup D. These are the first G protein sequences of non A/non-B APVs to be determined. PMID- 11038386 TI - Equine infectious anaemia virus proteins with epitopes most frequently recognized by cytotoxic T lymphocytes from infected horses. AB - Efficacious lentiviral vaccines designed to induce cytotoxic T lymphocytes (CTL) in outbred populations with a diverse repertoire of MHC class I molecules should contain or express multiple viral proteins. To determine the equine infectious anaemia virus (EIAV) proteins with epitopes most frequently recognized by CTL from seven horses infected for 0.5 to 7 years, retroviral vector-transduced target cells expressing viral proteins were used in CTL assays. Gag p15 was recognized by CTL from 100% of these infected horses. p26 was recognized by CTL from 86%, SU and the middle third of Pol protein were each recognized by 43%, TM by 29%, and S2 by 14%. Based on these results, it is likely that a construct expressing the 359 amino acids constituting p15 and p26 would contain epitopes capable of stimulating CTL in most horses. PMID- 11038387 TI - Simian immunodeficiency virus resistance of macaques infused with interferon beta engineered lymphocytes. AB - To test the in vivo anti-simian immunodeficiency virus (SIV) efficacy of interferon (IFN)-beta-engineered lymphocytes, peripheral blood lymphocytes harvested from two uninfected macaques were transduced with a retroviral vector carrying a constitutively expressed IFN-beta gene and reinfused, resulting in approximately 1 IFN-beta-transduced cell out of 1000 circulating cells. The gene modified cells were well tolerated and could be detected for at least 74 days without causing any apparent side effects. These two animals together with three untreated control macaques were then infected with SIVmac251. The two IFN-beta infused macaques are in good health, 478 days after infection, with a reduced plasma virus load and sustained numbers of CD4(+) and CD8(+) cells. Throughout the study, the proportion of IFN-beta-transduced cells has been maintained. Of the three control macaques, two were characterized by a high plasma virus load and a decrease in CD4(+) cells. One was moribund and was sacrificed 350 days after infection and the other now has fewer than 100 circulating CD4(+) cells/ml. Unexpectedly, the third control macaque, which, like the two IFN-beta-infused animals, had a low plasma virus load and a maintenance of CD4(+) and CD8(+) cell number, was characterized by a permanent level of serum IFN-beta, of unknown origin, already present before SIV infection. Although no definite conclusion can be made in view of the limited number of animals, these data indicate that further exploration is warranted of an IFN-beta-based anti-human immunodeficiency virus gene therapy. PMID- 11038388 TI - Cell-free synthesis of poliovirus: 14S subunits are the key intermediates in the encapsidation of poliovirus RNA. AB - In a cell-free system of uninfected HeLa cells, programmed with poliovirus RNA, extraneous radiolabelled 14S subunits assembled with endogenous 14S subunits and interacted with newly synthesized RNA to form virions (160S). This result suggests that 14S subunits are the key intermediates in the encapsidation of poliovirus RNA. PMID- 11038389 TI - Quantitative analysis of viral RNA kinetics in coxsackievirus B3-induced murine myocarditis: biphasic pattern of clearance following acute infection, with persistence of residual viral RNA throughout and beyond the inflammatory phase of disease. AB - Although the association remains controversial, enteroviruses have been implicated in the aetiology of several chronic diseases of humans. To further understand the mechanism of enterovirus persistence and its relationship to organ pathology, virus infectivity and viral RNA kinetics in the heart and other target organs during acute and persistent phases of murine coxsackievirus B3 infection were investigated. These studies revealed a biphasic pattern of virus clearance. Thus, there was a rapid but incomplete clearance of viral RNA from the myocardium following the acute phase of virus replication, which paralleled the elimination of virus infectivity. The mean half-life of viral RNA between days 5 and 14 post infection (p.i.) was 13.4 h. In contrast, a much slower rate of decline in viral RNA levels was observed during the post-infectious inflammatory phase of myocarditis. The mean half-life of viral RNA between days 14 and 90 p.i. was 14.1 days. Viral RNA persisted in the myocardium beyond the resolution of inflammation and was still detectable in a proportion of animals 90 days after infection. Clearance of viral RNA from other target organs occurred more rapidly, but the rate of clearance was largely independent of the level of viral RNA present during the acute phase of infection. Thus, while antiviral immune responses effectively eliminated infectious virus, clearance of residual viral RNA from the myocardium and other target organs was significantly delayed, despite a prolonged inflammatory response. These findings suggest that clearance of persistent enterovirus infection requires mechanisms different from those responsible for the elimination of virus infectivity. PMID- 11038390 TI - Construction of a full-length infectious cDNA clone of swine vesicular disease virus strain NET/1/92 and analysis of new antigenic variants derived from it. AB - The Dutch swine vesicular disease virus (SVDV) isolate NET/1/92 was one of the first isolates belonging to a new SVDV antigenic group. This strain was completely sequenced and was shown to have 93% similarity with the UKG/27/72 isolate. To enable antigenicity, replication, maturation and pathogenicity studies of NET/1/92, an infectious full-length cDNA clone, designated pSVD146, was prepared. The in vitro and in vivo biological properties of the virus derived from pSVD146 were studied by analysing antigenicity, plaque morphology, growth curves and virulence in pigs. The epitopes of newly prepared monoclonal antibodies were roughly mapped by fusion-PCR. Fine mapping of epitopes at the amino acid level was achieved by introducing single amino acid mutations in pSVD146. Two new amino acids important in epitope formation were located in VP1; one was mapped in the C-terminal end and the second is thought to be located in the H-I loop. Growth curve and plaque sizes in vitro were similar between virus derived from pSVD146 and the parent wild-type virus. In virulence studies in pigs, the lesions score, neutralization titres and the seroconversion rates were comparable between virus derived from pSVD146 and the parent strain. Since virus derived from pSVD146 had the same biological properties as the parent strain NET/1/92, the full-length infectious cDNA clone pSVD146 will be very useful in studies of the antigenicity, virulence, pathogenesis, maturation and replication of SVDV. PMID- 11038391 TI - Proteolytic processing at a novel cleavage site in the N-terminal region of the tomato ringspot nepovirus RNA-1-encoded polyprotein in vitro. AB - Tomato ringspot nepovirus RNA-1-encoded polyprotein (P1) contains the domains for the putative NTP-binding protein, VPg, 3C-like protease and a putative RNA dependent RNA polymerase in its C-terminal region. The N-terminal region of P1, with a coding capacity for a protein (or a precursor) of 67 kDa, has not been characterized. Using partial cDNA clones, it is shown that the 3C-like protease can process the N-terminal region of P1 at a novel cleavage site in vitro, allowing the release of two proteins, X1 (located at the N terminus of P1) and X2 (located immediately upstream of the NTB domain). P1 precursors in which the protease was inactive or absent were not cleaved by exogenously added protease, suggesting that P1 processing was predominantly in cis. Results from site directed mutagenesis of putative cleavage sites suggest that dipeptides Q(423)/G and Q(620)/G are the X1-X2 and X2-NTB cleavage sites, respectively. The putative X1 protein contains a previously identified alanine-rich sequence which is present in nepoviruses but not in the related comoviruses. The putative X2 protein contains a region with similarity to the comovirus 32 kDa protease co factor (the only mature protein released from the N terminus of comovirus P1 polyproteins) and to the corresponding region of other nepovirus P1 polyproteins. These results raise the possibility that the presence of two distinct protein domains in the N-terminal part of the P1 polyprotein may be a common feature of nepoviruses. PMID- 11038392 TI - Characterization of VPg and the polyprotein processing of cocksfoot mottle virus (genus Sobemovirus). AB - The polyprotein of Cocksfoot mottle virus (CfMV; genus SOBEMOVIRUS:) is translated from two overlapping open reading frames (ORFs) 2a and 2b by a -1 ribosomal frameshifting mechanism. In this study, a 12 kDa protein was purified from viral RNA-derived samples that appears to correspond to the CfMV genome linked protein (VPg). According to the determined N-terminal amino acid sequence, the VPg domain is located between the serine proteinase and replicase motifs and the N terminus of VPg is cleaved from the polyprotein between glutamic acid and asparagine residues. Western blot analysis of infected plant material showed that the polyprotein is processed at several additional sites. An antiserum against the ORF 2a product recognized six distinct proteins, whereas, of these, the VPg antiserum clearly recognized only a 24 kDa protein. This indicates that the fully processed 12 kDa VPg detected in viral RNA-derived samples is a minor product in infected plants. An antiserum against the ORF 2b product recognized a 58 kDa protein, which indicates that the fully processed replicase is entirely or almost entirely encoded by ORF 2b. The origin of the detected cleavage products and a proposed polyprotein processing model are discussed. PMID- 11038393 TI - Mechanical transmission of Potato leafroll virus. AB - Like typical luteoviruses, Potato leafroll virus (PLRV) cannot be transmitted mechanically by rubbing plants with solutions containing virus particles. However, PLRV was found to be mechanically transmissible from extracts of plants that had been inoculated by viruliferous aphids and then post-inoculated with Pea enation mosaic virus-2 (PEMV-2). Unlike the asymptomatic infections induced by either virus alone, double infections in Nicotiana benthamiana induced necrotic symptoms with some line patterning and vein yellowing. Infective PLRV was recovered from a purified virus preparation by inoculating plants mechanically with purified virus particles mixed with PEMV-2. Similarly, Beet mild yellowing virus was readily transmitted mechanically from mixtures containing PEMV-2. PLRV was also transmissible from mixtures made with extracts of plants infected with Groundnut rosette virus, although less efficiently than from mixtures containing PEMV-2. This novel means of transmitting PLRV, and perhaps other poleroviruses, should prove very useful in a number of fields of luteovirus research. PMID- 11038394 TI - Natural recombination between Tomato yellow leaf curl virus-is and Tomato leaf curl virus. AB - The complete genome sequences (2791 and 2793 nt) of isolates of Tomato yellow leaf curl virus-Is (TYLCV-Is) from Spain (SP72/97) and Portugal (Port2/95) were determined. These isolates are closely related to TYLCV-Is isolates reported in Japan (Japan-A and Japan-S) and Israel (Israel/Mild). Comparison of all sequenced isolates of TYLCV-Is showed that part of the genome comprising the intergenic region and the 5'-end of the rep gene of the Iran and Israel isolates was not closely related to that of other isolates. Phylogenetic analyses suggest that the Israel and Iran isolates may have chimeric genomes that have arisen by recombination between TYLCV-Is-like and tomato leaf curl virus (ToLCV)-like ancestors. The TYLCV-Is donors of the Iran and the Israel genomes were closely related to each other and to other known TYLCV-Is isolates. However, the ToLCV donors differed from each other, although both were related to ToLCV isolates from India (Bangalore-2 and Bangalore-4). PMID- 11038395 TI - The inhibitors of apoptosis of Epiphyas postvittana nucleopolyhedrovirus. AB - In this study, four inhibitor of apoptosis genes (iaps) in the genome of Epiphyas postvittana nucleopolyhedrovirus (EppoMNPV) that are homologous to iap-1, iap-2, iap-3 and iap-4 genes of other baculoviruses have been identified. All four iap genes were sequenced and the iap-1 and iap-2 genes were shown to be functional inhibitors of apoptosis. The iap-1, iap-2 and iap-3 genes contain two baculovirus apoptosis inhibitor repeat motifs and a C(3)HC(4) RING finger-like motif. The activity of the iap genes was tested by transient expression in Spodoptera frugiperda (Sf-21) cells treated with the apoptosis-inducing agents actinomycin D, cycloheximide, anisomycin, tumour necrosis factor-alpha and UV light. The iap 2 gene prevented apoptosis induced by all agents tested, indicating activity towards a conserved component(s) of multiple apoptotic pathways. However, the iap 2 gene was unable to function in the absence of a gene immediately upstream of iap-2 that has homology to the orf69 gene of Autographa californica MNPV. The use of a CMV promoter rescued the apoptosis inhibition activity of the iap-2 gene, indicating that the upstream orf69 homologue is associated with expression of iap 2. The iap-1 gene was able to delay the onset of apoptosis caused by all of the induction agents tested but, unlike iap-2, was unable to prevent the development of an apoptotic response upon prolonged exposure of cells to the apoptosis induction agents. No anti-apoptotic activity was observed for the iap-3 and iap-4 genes of EppoMNPV. PMID- 11038396 TI - PrP(C) expression in the peripheral nervous system is a determinant of prion neuroinvasion. AB - Transmissible spongiform encephalopathies are often propagated by extracerebral inoculation. The mechanism of spread from peripheral portals of entry to the central nervous system (neuroinvasion) is complex: while lymphatic organs typically show early accumulation of prions, and B-cells and follicular dendritic cells are required for efficient neuroinvasion, actual entry into the central nervous system occurs probably via peripheral nerves and may utilize a PrP(C) dependent mechanism. This study shows that transgenic mice overexpressing PrP(C) undergo rapid and efficient neuroinvasion upon intranerval and footpad inoculation of prions. These mice exhibited deposition of the pathological isoform of the prion protein (PrP(Sc)) and infectivity in specific portions of the central and peripheral sensory pathways, but almost no splenic PrP(Sc) accumulation. In contrast, wild-type mice always accumulated splenic PrP(Sc), and had widespread deposition of PrP(Sc) throughout the central nervous system even when prions were injected directly into the sciatic nerve. These results indicate that a lympho-neural sequence of spread occurs in wild-type mice even upon intranerval inoculation, while overexpression of PrP(C) leads to substantial predilection of intranerval over lymphoreticular spread. The rate of transport of infectivity in peripheral nerves was ca. 0.7 mm per day, and prion infectivity titres of sciatic nerves were much higher in tga20 than in wild-type mice, suggesting that overexpression of PrP(C) modulates the capacity for intranerval transport. PMID- 11038397 TI - Development of an improved technique for the perfusion of the isolated caudal lobe of sheep liver. AB - The study was designed to develop an improved technique for perfusing the isolated caudal lobe of sheep liver. Twenty caudal lobes were perfused for 3-4 h, in a non-recirculating mode, with Krebs-Henseleit bicarbonate buffer. The perfusion system was designed to give a constant flow. The hepatic viability and functional normality of the perfused lobe were assessed by measuring the perfusion flow rate, pH, K+ efflux, O2 uptake, substrate uptake, gluconeogenesis from propionate and amino acids, and ureagenesis from ammonia and amino acids. Liver tissue was sampled for histological examination, as well as for the determination of liver glycogen and wet : dry weight ratio. The perfusion flow rate and pH were both stable throughout the perfusion. The potassium concentration in the effluent perfusate did not increase during the perfusion, suggesting that there was no loss of viability or hypoxia. The perfused lobe extracted more than 50% of the O2 supply. The rate of oxygen consumption was comparable to the rate reported in vivo. The initial glycogen content was reduced by about 40% after 4 h perfusion. The wet : dry weight ratio was 3.6, consistent with the absence of tissue oedema. Urea production was stimulated when NH4Cl (0.3 mM) was added to the medium but there was no significant increase in urea release when alanine (0.15 mM), glutamine (0.2 mM) or lysine (0.2 mM) was added. Urea production, however, increased by about 171% when a physiological mixture of amino acids was added. Propionate (0.5 mM), alanine and glutamine stimulated glucose production but not lysine or the complete amino acid mixture. Glutamine release was lower than that reported in the rat liver. Changing the direction of flow also revealed an apparent difference between livers from sheep and rats in their metabolism of ammonia. The improved technique offers a simple practical and inexpensive approach to many problems in ruminant physiology and nutritional biochemistry. PMID- 11038398 TI - Possible contribution of central gamma-aminobutyric acid receptors to resting vascular tone in freely moving rats. AB - Previous studies have shown that central administration of GABA (gamma aminobutyric acid), an inhibitory neurotransmitter, preferentially reduces hindquarters and carotid vascular resistances but not renal and coeliac vascular resistances in conscious rats. This study tested the hypothesis that these preferential actions of central GABA receptors are related to differences between vessels in resting autonomic vascular tone in freely moving rats. Rats were chronically implanted with intracisternal cannulas and/or electromagnetic probes to measure regional blood flows. In response to GABA administration, the changes in vascular resistance (arterial blood pressure/regional blood flow) of the hindquarters (n = 23) and carotid (n = 12) vascular beds were significantly and negatively correlated with basal vascular resistance. No such relationship was found for the renal (n = 21), coeliac (n = 13) and superior mesenteric (n = 23) vascular beds. This finding indicates that the responsiveness to GABA of brainstem pathways controlling the hindquarters and carotid vascular beds co varies with resting resistance in hindquarters and carotid vessels. A similar analysis was performed, correlating the ongoing vascular resistance of each vessel with its response to ganglionic blockade by chlorisondamine. In this case, a significant negative correlation was also found for the hindquarters (n = 26) and carotid (n = 15) vascular beds, but not for the coeliac (n = 17) or superior mesenteric (n = 19) vessels. Together, these findings suggest that central GABA receptors accessible from the cisterna magna preferentially affect two vascular beds which, in the freely moving rat, show resting autonomic vascular tone. PMID- 11038399 TI - Regional distribution of potassium currents in the rabbit pulmonary arterial circulation. AB - The response of pulmonary arteries to hypoxia varies as a function of vessel diameter. Small intrapulmonary resistance arteries are thought to be the main site of hypoxic pulmonary vasoconstriction (HPV), with hypoxia causing minimal contraction or even dilatation in large, conduit vessels. This has been proposed to reflect a differential distribution of morphologically and electrophysiologically distinct pulmonary artery smooth muscle (PASM) cells. We investigated longitudinal heterogeneity in smooth muscle cells isolated from five regions of the rabbit pulmonary vasculature and could find no evidence of morphological heterogeneity at the level of the light microscope. PASM cells from main (8 mm outer diameter) and branch (5 mm) arteries and large ( 400 m) intrapulmonary arteries (IPA) were similar in shape and size, as indicated by cell capacitance (25 pF). PASM cells from medium (200-400 m) and small ( 200 m) IPA were significantly smaller (15 pF), but had the same classical spindle shape. Cells from all five regions also had similar resting membrane potentials and displayed voltage-activated K+ currents of similar amplitude when recorded in standard physiological solution. Longitudinal heterogeneity in K+ current became apparent when tetraethylammonium ions (TEA; 10 mM) and glibenclamide (10 M) were added. The remaining delayed rectifier current (IK(V)) doubled in amplitude upon moving down the pulmonary arterial tree from the main artery (9 pA pF-1 at 40 mV) to the large IPA (17 pA pF-1), but remained constant throughout the intrapulmonary vasculature. The O2-sensitive, non-inactivating K+ current (IK(N)) showed a similar trend, but was significantly reduced in the smallest IPA, where its amplitude was comparable with the main artery. Thus the IK(N)/IK(V) ratio was relatively constant, at around 0.14, from the main pulmonary artery to medium IPA, but fell by 50% in the smallest vessels. The amplitude of the TEA-sensitive K+ current was similar (16 pA pF-1 at 40 mV) at all levels of the pulmonary arterial tree, except in the medium sized vessels where it was 50% smaller. These variations in K+ current expression correlate with reported variations in sensitivity to hypoxia and may contribute to the regional heterogeneity of HPV in the rabbit lung. PMID- 11038400 TI - Hypotonic swelling increases L-type calcium current in smooth muscle cells of the human stomach. AB - The purpose of the present study was to characterize the Ca2+ channels in smooth muscle cells from human stomach and to examine the effects of osmotic swelling on the channel activity. Ca2+ channel current with either Ca2+ or Ba2+ as charge carrier was recorded from freshly isolated smooth muscle cells using the conventional whole-cell patch clamp technique. The degree of cell swelling as a result of hypotonic challenge was monitored using a video image analysis system. The changes in intracellular Ca2+ concentration ([Ca2+]i) were measured by microfluorimetry. The pharmacological and voltage activation profile suggests a typical dihydropyridine-sensitive L-type Ca2+ current. Cell swelling, induced by hypotonic challenge, enhanced the amplitude of currents through L-type Ca2+ channels without significant effects on steady-state voltage dependency. After treatment with the L-type Ca2+ channel agonist Bay K 8644 (0.1-2 microM), no further significant increase in calcium channel current or corresponding [Ca2+]i transients were provoked by the swelling. The above results demonstrated that the presence of L-type Ca2+ current in smooth muscle cells of the human stomach and the augmentation of the current are closely associated with the volume increase resulting from hypotonic swelling. PMID- 11038401 TI - Differences in the calcium-handling response of isolated rat and guinea-pig cardiomyocytes to metabolic inhibition: implications for cell damage. AB - Species differences in response to hypoxic damage have been observed in studies using whole hearts. The aims of this study were to determine whether (i) species differences in response to simulated hypoxia could be detected at the level of the single myocyte, and (ii) there were any interspecies differences in the Ca2+ handling properties of the cells. Ventricular myocytes were isolated from hearts of adult rats and guinea-pigs and electrically stimulated on the stage of a fluorescence microscope. Cell length was measured using an edge-tracking device, and total intracellular [Ca2+] ([Ca2+]i) determined using indo-1. Cells were exposed to metabolic inhibition (MI) (2.5 mM NaCN and no glucose) to simulate hypoxia followed by washout of CN and re-addition of glucose ('reperfusion'). Following exposure to MI, rat cells underwent rigor contracture in 18.8+/-0.8 min (n = 80 cells), whereas the time was longer for guinea-pig cells (32.9+/-1.2 min, n = 83) (P<0.001). If cells were reperfused after 1-5 min in rigor, then rat cells showed improved morphological recovery compared with guinea-pig cells (P< 0.05); thereafter recovery decreased with increasing time spent in rigor, and was similar in both groups. In indo-1 loaded cells, [Ca2+]i was significantly increased in cells from both species at the end of MI; however, the actual increase was much higher in guinea-pig cells. Upon reperfusion, [Ca2+]i recovered fully in rat cells, but in guinea-pig cells there was no significant decrease. The restoration of [Ca2+]i to normal levels in rat cells following MI was associated with improved contractile recovery compared with guinea-pig cells. We conclude that rat cells are more resistant to effects of MI than are guinea-pig cells; this may be related to species differences in Ca2+ handling during and following exposure to MI. PMID- 11038402 TI - Nerve-evoked secretion of immunoglobulin A in relation to other proteins by parotid glands in anaesthetized rat. AB - Secretion of fluid and proteins by salivary cells is under the control of parasympathetic and sympathetic autonomic nerves. In a recent study we have shown that, in the rat submandibular gland, autonomic nerves can also increase the secretion of IgA, a product of plasma cells secreted into saliva as SIgA (IgA bound to Secretory Component, the cleaved poly-immunoglobulin receptor). The present study aimed to determine if parotid secretion of SIgA is increased by autonomic nerves and to compare SIgA secretion with other parotid proteins stored and secreted by acinar and ductal cells. Assay of IgA in saliva evoked by parasympathetic nerve stimulation immediately following an extended rest period under anaesthesia indicated that it had been secreted into intraductal saliva in the absence of stimulation during the rest period. The mean rate of unstimulated IgA secretion (2.77+/-0.28 microg min(-1) g(-1)) and the 2.5-fold increase in IgA secretion evoked by parasympathetic stimulation were similar to results found previously in the rat submandibular gland. Sympathetic nerve stimulation increased SIgA secretion 2.7-fold, much less than in the submandibular gland. SDS PAGE and Western blot analysis with anti-IgA and anti-Secretory Component antibodies confirmed that SIgA was the predominant form of IgA in saliva. Acinar derived amylase and ductal-derived tissue kallikrein were more profoundly increased by parasympathetic and particularly sympathetic stimulation than SIgA. Overall, the results of the present study indicate that SIgA forms a prominent component of unstimulated parotid salivary protein secretion and that its secretion is similarly increased by stimulation of either autonomic nerve supply. The secretion of other parotid salivary proteins that are synthesized and stored by acinar or ductal cells is upregulated to a much greater extent by parasympathetic and particularly sympathetic stimulation. PMID- 11038403 TI - Ageing modulates some aspects of the non-specific immune response of murine macrophages and lymphocytes. AB - The deterioration of the immune system with ageing, which leads to an increased morbidity and mortality from infections, appears to be related to decreases in specific lymphocyte functions. However, the alteration of non-specific immunity is a more controversial subject. Our purpose was to investigate the age-related changes of different functions of the non-specific immune response in peritoneal macrophages (adherence to tissues, mobility directed to a chemical gradient from an infectious focus or chemotaxis, phagocytosis of foreign agents and destruction of these agents by superoxide anion production) and in lymphocytes (adherence and chemotaxis) from peritoneum, axillary lymph nodes, spleen and thymus. We used young (12 weeks), adult (22 weeks), mature (48 weeks) and old (72 weeks) female BALB/c mice. The adherence capacity of macrophages and lymphocytes was greater in adult and old mice than in young animals. The chemotaxis of macrophages showed higher values in cells from young mice than in those from adult mice, increasing again in macrophages from mature and old animals. A similar behaviour was shown by phagocytosis, which reached its highest values in old animals. Anion superoxide production increased with age and again the highest values were obtained in the oldest mice. Conversely, chemotaxis of lymphocytes was higher in the adult and mature animals than in the young and old animals. We conclude that, although there is a decrease in lymphocyte chemotaxis in old animals, the non specific immune response of macrophages instead of decreasing, may increase in aged mice with respect to the values seen in adult mice. PMID- 11038404 TI - The preoptic area in the hypothalamus is the source of the additional respiratory drive at raised body temperature in anaesthetised rats. AB - In mammals that use the ventilatory system as the principal means of increasing heat loss, raising body temperature causes the adoption of a specialised breathing pattern known as panting and this is mediated by the thermoregulatory system in the preoptic area of the hypothalamus. In these species an additional respiratory drive is also present at raised body temperature, since breathing can reappear at low Pa,CO2 levels, when stimulation of chemoreceptors is minimal. It is not known whether the preoptic area is also the source of this additional drive. Rats do not pant but do possess this additional respiratory drive at raised body temperatures. We have therefore tested whether the preoptic area of the hypothalamus is the source of this additional respiratory drive in rats. Urethane anaesthesia and hyperoxia were used in eleven rats to minimise behavioural and chemical drives to breathe. The presence of the additional respiratory drive was indicated if rhythmic diaphragmatic EMG activity reappeared during hypocapnia (a mean Pa,CO2 level of 21+/-2 mm Hg, n = 11), induced by mechanical ventilation. The additional respiratory drive was absent at normal body temperature (37?C). When the temperature of the whole body was raised using an external source of radiant heat, the additional respiratory drive appeared at 40.6+/-0.5 degrees C (n = 3). In two further rats this drive was induced at normal body temperature by localised warming in the preoptic area of the intact hypothalamus. The additional respiratory drive appeared at similar temperatures to those in control rats in three rats following isolation of the hypothalamus from more rostral areas of the brain. In contrast, the additional respiratory drive failed to appear at these temperatures in three rats after isolating the hypothalamus from the caudal brainstem, by sectioning pathways medial to the medial forebrain bundle. Since the preoptic area is known to contain thermoreceptors and to receive afferents from peripheral thermoreceptors, the results show that this area is also the source of the additional respiratory drive at raised body temperature in anaesthetised rats. PMID- 11038405 TI - Equine uteroplacental metabolism at mid- and late gestation. AB - Uptakes of oxygen, glucose and lactate by the gravid uterus, fetus and uteroplacental tissues were measured in chronically catheterized pregnant ponies and their fetuses at mid- and late gestation (term 335 days). Rates of O2 uptake by the gravid uterus, fetus and uteroplacental tissues were significant at both gestational ages and were 2- to 3-fold higher in late gestation than the mid gestation values of 3338+/-794, 1352+/-258 and 2035 +/- 602 micromol min(-1), respectively (n = 4). Similarly, there were significant uptakes of glucose by the gravid uterus, fetus and uteroplacental tissues at both mid- and late gestation. However, unlike O2 uptake, glucose uptake by the uterus and uteroplacental tissues did not increase between mid- and late gestation. No significant uptakes or outputs of lactate were observed by the uterus or uteroplacental tissues at either gestational age, although there was a significant umbilical uptake of lactate in late but not mid-gestation. There was no change in the distribution of uterine O2 uptake between the fetus and uteroplacental tissues with increasing gestational age. The uteroplacental tissues accounted for about 50 % of the uterine O2 uptake at both gestational ages. In contrast, the proportion of the uterine glucose uptake used by the uteroplacental tissues decreased from 73.2+/ 2.1 % (n = 5) at mid-gestation to 61.1+/-1.9 % (n = 4, P<0.02) in late gestation. The gestational changes in uteroplacental carbohydrate metabolism in the mare differ from those seen in the ewe and may have important consequences for the duration and outcome of pregnancy in the mare. PMID- 11038406 TI - Left ventricular diastolic filling and cardiovascular functional capacity in older men. AB - We investigated anaerobic threshold (< theta(L)) gas exchange kinetics and maximal oxygen uptake (VO2,max) among older men with reduced left ventricular end diastolic filling (LVDF). Ten men (mean age, 73 years) with LVDF impairment and low fitness, but without other cardiovascular dysfunction were studied. Treatments compared to control included: 5 days, high intensity exercise training protocol; 5 days, calcium channel blockade (240 mg verapamil); 21 days, detraining/washout; and 5 days, combined treatments. Results indicated no changes in resting left ventricular systolic function with any treatment. Significant resting diastolic function changes included increased early:late flow velocity (control, 0.87; training, 1.28; verapamil, 1.32), and a decreased isovolumic relaxation time (control, 0.10 s; training, 0.08 s; verapamil, 0.08 s). The combined treatments were not additive. Sub-threshold oxygen uptake kinetics (tauVO2, s) were significantly faster following either training or verapamil (tauVO2,control, 62+/-12; tauVO2,training, 44+/-9; tauVO2,verapamil, 48+/-10) and combined treatments (tauVO2, 41+/- 8). V O2,max (ml kg(-1) min(-1)) was significantly increased (control, 21.8+/-2.2; training, 27.3+/-2.2; verapamil, 25.2+/-3.4; combined treatments, 26.9+/-2.3). Increasing ventricular preload with either exercise training or calcium channel blockade was coincident with faster tauVO2 and increased VO2,max. PMID- 11038407 TI - Cardiovascular effects of 8 h of isocapnic hypoxia with and without beta-blockade in humans. AB - This study seeks to confirm the progressive changes in cardiac output and heart rate previously reported with 8 h exposures to constant hypoxia, and to examine the role of sympathetic mechanisms in generating these changes. Responses of ten subjects to four 8 h protocols were compared: (1) air breathing with placebo; (2) isocapnic hypoxia (end-tidal PO2 = 50 mm Hg) with placebo; (3) isocapnic hypoxia with beta-blockade; and (4) air breathing with beta -blockade. Regular measurements of heart rate and cardiac output (using ultrasonography and N2O rebreathing techniques) were made with subjects seated in the upright position. The sensitivity of heart rate to rapid variations in hypoxia (GHR) and heart rate in the absence of hypoxia were measured at times 0, 4 and 8 h. No significant progressive effect of hypoxia on cardiac output was detected. There was a gradual rise in heart rate with hypoxia of 11+/-2 beats min(-1) in the placebo protocol and of 10+/-2 beats min(-1) in the beta-blockade protocol over 8 h, compared to the air breathing protocols. The rise in heart rate was progressive (P<0.001) and accompanied by progressive increases in both GHR (P<0.001) and heart rate measured in the absence of hypoxia (P<0.05). No significant effect of beta blockade was detected on any of these progressive changes. We conclude that sympathetic mechanisms that act via beta -receptors play little role in the progressive changes in heart rate observed over 8 h of moderate hypoxia. PMID- 11038408 TI - Selective long-term electrical stimulation of fast glycolytic fibres increases capillary supply but not oxidative enzyme activity in rat skeletal muscles. AB - Glycolytic fibres in rat extensor digitorum longus (EDL) and tibialis anterior (TA) were selectively activated, as demonstrated by glycogen depletion, by indirect electrical stimulation via electrodes implanted in the vicinity of the peroneal nerve using high frequency (40 Hz) trains (250 ms at 1 Hz) and low voltage (threshold of palpable contractions). This regime was applied 10 times per day, each bout being of 15 min duration with 60 min recovery, for 2 weeks. Cryostat sections of muscles were stained for alkaline phosphatase to depict capillaries, succinate dehydrogenase (SDH) to demonstrate oxidative fibres, and periodic acid-Schiff reagent (PAS) to verify glycogen depletion. Specific activity of hexokinase (HK), 6-phosphofructokinase, pyruvate kinase, glycogen phosphorylase and cytochrome c oxidase (COX) were estimated separately in homogenates of the EDL and the predominantly glycolytic cortex and oxidative core of the TA. Stimulation increased the activity of HK but not that of oxidative enzymes in fast muscles. Comparison of changes in oxidative capacity and capillary supply showed a dissociation in the predominantly glycolytic TA cortex. Here, COX was 3.9+/-0.68 microM min(-1) (g wet wt)-1 in stimulated muscles compared with 3.7+/-0.52 microM min(-1) (g wet wt)-1 in contralateral muscles (difference not significant), while the percentage of oxidative fibres (those positively stained for SDH) was also similar in stimulated (14.0+/-2.8 %) and contralateral (12.2 +/-1.9 %) muscles. In contrast, the capillary to fibre ratio was significantly increased (2.01+/-0.12 vs. 1.55+/-0.04, P<0.01). We conclude that capillary supply can be increased independently of oxidative capacity, possibly due to haemodynamic factors, and serves metabolite removal to a greater extent than substrate delivery. PMID- 11038409 TI - Angiotensin-converting enzyme genotype affects the response of human skeletal muscle to functional overload. AB - The response to strength training varies widely between individuals and is considerably influenced by genetic variables, which until now, have remained unidentified. The deletion (D), rather than the insertion (I), variant of the human angiotensin-converting enzyme (ACE) genotype is an important factor in the hypertrophic response of cardiac muscle to exercise and could also be involved in skeletal muscle hypertrophy - an important factor in the response to functional overload. Subjects were 33 healthy male volunteers with no experience of strength training. We examined the effect of ACE genotype upon changes in strength of quadriceps muscles in response to 9 weeks of specific strength training (isometric or dynamic). There was a significant interaction between ACE genotype and isometric training with greater strength gains shown by subjects with the D allele (mean +/- S.E.M.: II, 9.0+/-1.7 %; ID, 17.6 +/-2.2 %; DD, 14.9+/-1.3 %, ANOVA, P 0.05). A consistent genotype and training interaction (ID DD II) was observed across all of the strength measures, and both types of training. ACE genotype is the first genetic factor to be identified in the response of skeletal muscle to strength training. The association of the ACE I/D polymorphism with the responses of cardiac and skeletal muscle to functional overload indicates that they may share a common mechanism. These findings suggest a novel mechanism, involving the renin-angiotensin system, in the response of skeletal muscle to functional overload and may have implications for the management of conditions such as muscle wasting disorders, prolonged bed rest, ageing and rehabilitation, where muscle weakness may limit function. PMID- 11038410 TI - Carbohydrate ingestion prior to exercise augments the exercise-induced activation of the pyruvate dehydrogenase complex in human skeletal muscle. AB - This study examined the effect of pre-exercise carbohydrate (CHO) ingestion on pyruvate dehydrogenase complex (PDC) activation, acetyl group availability and substrate level phosphorylation (glycogenolysis and phosphocreatine (PCr) hydrolysis) in human skeletal muscle during the transition from rest to steady state exercise. Seven male subjects performed two 10 min treadmill runs at 70 % maximum oxygen uptake (VO2,max), 1 week apart. Each subject ingested 8 ml (kg body mass (BM))-1 of either a placebo solution (CON trial) or a 5.5 % CHO solution (CHO trial) 10 min before each run. Muscle biopsy samples were obtained from the vastus lateralis at rest and immediately after each trial. Muscle PDC activity was higher at the end of exercise in the CHO trial compared with the CON trial (1.78+/-0.18 and 1.27+/-0.16 mmol min(-1) (kg wet matter (WM))(-1), respectively; P 0.05) and this was accompanied by lower acetylcarnitine (7.1+/ 1.2 and 9.1+/-1.1 mmol kg(-1) (dry matter (DM))(-1) in CHO and CON, respectively; P<0.05) and citrate concentrations (0.73+/-0.05 and 0.91+/-0.10 mmol (kg DM)(-1) in CHO and CON, respectively; P<0.05). No difference was observed between trials in the rates of muscle glycogen and PCr breakdown and lactate accumulation. This is the first study to demonstrate that CHO ingestion prior to exercise augments the exercise-induced activation of muscle PDC and reduces acetylcarnitine accumulation during the transition from rest to steady-state exercise. However, those changes did not affect the contribution of substrate level phosphorylation to ATP resynthesis. PMID- 11038411 TI - [Pierre Masson (1880-1959): a towering figure in tumor histopathology]. PMID- 11038412 TI - [The different meanings of the word stratification]. AB - In clinical research, stratification has three different meanings. In its first meaning, it describes the natural distribution of patients into subgroups, for instance patients may be stratified by stage. In its second meaning, stratification controls the random allocation of the treatments in a clinical trial, for instance the randomisation can be stratified on stage. Lastly, in its third meaning, stratification of a trial analysis consists into taking into account patients' characteristics in the analysis. This three meanings will be illustrated with a basic example. PMID- 11038413 TI - [p53 activation by PI-3K family kinases after DNA double-strand breaks]. AB - p53 plays a central role in the cellular response to DNA double-strand breaks (DSBs), and to DNA damage in general. The protein kinases ATM, ATR and DNA-PK detect DSBs and transmit this information to p53 by phosphorylation. This phosphorylation dissociates p53 from its negative regulator, mdm2. p53 then undergoes further modification and activates transcription of the genes responsible for cell cycle arrest. In certain circumstances, p53 also activates transcription of the genes responsible for apoptosis. The dysfunction of this cascade of events is oncogenic, with P53 itself being the most commonly mutated gene in malignant cells, although mutations in both the DNA damage sensors and cell cycle checkpoint and apoptosis effectors are frequent. A more complete understanding of p53 and the proteins it interacts with may allow the development of new cancer treatments. PMID- 11038415 TI - ["Psychogenesis" of cancer: between myths, misuses and reality]. AB - Since a long time, hypothesis of links between psychological factors and cancer, have been established in our culture. So far, numerous researches have tempted to indicate stress, coping facing the disease, depression or "type C" personality as factors participating to the onset and/or the course of the cancer. A review of those studies, mainly retrospective, has mostly brought debated results, as well as prospective researches including large sample of population or people awaiting a diagnosis; therefore making old-fashioned every area strictly "psychogenetic" of cancer at first sight. Explicative indirect hypothesis are suggested by the psychoneuroendocrinoimmunology. Various researches in this field proved that external factors such as stress, depression or social support have significative influences on components of the immune system which in turn influence the onset and/or the course of the cancer. The links between psychological factors and cancer are extremely complex, bringing numerous biological, psychological or even sociological systems in interactions. Then psychoneuroendocrinoimmunology constitutes an early interdisciplinary way of mediation, capable of account for the connections between psychology and cancer. PMID- 11038414 TI - [Chemotherapy of metastatic breast cancer]. AB - The most powerful prognostic factor in the treatment of metastatic breast cancer continues to be the response to induction chemotherapy. The range of drugs which are widely used in the treatment of advanced disease, the anthracyclines, the taxanes and vinorelbine, have all shown interesting activity in terms of their ability to obtain both high response rates and long duration of response. The anthracyclines, doxorubicin (DX) and epiadriamycin (EPI) constitute the established reference agents in the treatment of metastatic disease, and combinations of these drugs with vinorelbine (VRB) and the taxanes, paclitaxel (PTX) and docetaxel (DCT), have produced major increases in objective response rates: PTX-DX (58%), DCT-EPI (69.4%), PTX-EPI (71.1%), VRB-DX (75%), VRB-EPI (77.1%). Suggestions for other combinations of chemotherapeutic agents which do not include anthracyclines available with well tolerated and effective drugs. The way forward after a response has been obtained remains an open question in which the limited efficacy of the available drugs and their cumulative toxicity needs to be balanced against the quality of life of patients during their disease. Defining the optimal strategy for the management of disease after induction treatment is a problem which needs to draw on the results of research, analysis of experience and insight into the needs of patients. PMID- 11038416 TI - [Complications and tolerance of heated intraperitoneal chemotherapy and cytoreductive surgery for peritoneal carcinomatosis: results of a phase I-II study of peritoneal carcinomatosis from different sources]. AB - Peritoneal carcinomatosis represents the terminal stage of adenocarcinomas of the gastrointestinal tract. A new treatment combining cytoreductive surgery and intraperitoneal chemotherapy-hyperthermia has been used with encouraging results. The purpose of this study was to report the complications associated with this treatment. Fourty procedures were carry out in 37 patients. Death occurred in 3 patients. Major medical complications were 13 pulmonary infections, 11 acute renal failure (with only 3 who needed dialysis) and 10 patients with neutropenia grade 3 and 4 toxicity. Intra-abdominal complications occurred in 16 patients (there were 11 anastomotic leak and/or bowel perforation, and 12 intra-abdominal infections). Some complications like secretory diarrhea or tubulopathia which were related to these treatment need further investigations. Six procedures were without any complications, 6 presented minor ones and 22 major complications. Adverse effects were relatively important with this new treatment strategy. This was maybe due to a learning process; there is no death and only one anastomotic leak in our last study including 30 patients with cytoreductive surgery and intraperitoneal chemotherapy-hyperthermia. PMID- 11038417 TI - [Huriet law: application in a regional cancer centre (Centre Leon-Berard, Lyon)]. AB - Clinical research is one of the main activities in cancer centres and is submitted in France to a specific law (named "loi Huriet") which includes good clinical practices. We are now conducting a general program of quality evaluation and improvement in the regional cancer centre of Lyon (centre Leon-Berard). Part of this program is an audit of the application of the Huriet law. Since no instrument exist for measuring this application, we have created a specific one, that attribute notation according to the different aspects of the law. Results show a good level of conformity but sometime non sufficient. There is no difference between the two studied years. Quality changes according to promoters (private or academic) and monitoring. Written procedures and specific training for the different actors are required to improve quality of clinical research with focus on the patient interest. PMID- 11038418 TI - [L'oncologie medicale : le point de vue des internes de specialites] PMID- 11038419 TI - [Cancer of the colon: an update] PMID- 11038420 TI - [Rectal cancers: an update] PMID- 11038421 TI - [Cancer of the canal anal: literature review] PMID- 11038422 TI - Cell block preparation by inverted filter sedimentation is useful in the differential diagnosis of atypical glandular cells of undetermined significance in ThinPrep specimens. AB - BACKGROUND: Atypical glandular cells of undetermined significance (AGUS) is not a well defined entity in the Bethesda system for reporting cervical cytology diagnoses. An AGUS diagnosis can reflect a variety of conditions ranging from the benign to the malignant. To help differentiate abnormalities associated with suspected AGUS in ThinPrep specimens, the authors devised a novel technique to prepare cell blocks that avoids disruption of glandular structures during preparation. METHODS: In the authors' technique, the fluid remaining after the preparation of ThinPrep slide was subjected to sedimentation in the inverted ThinPrep filter cylinder that was used for slide preparation. Sediment that was formed after filtration was completed was collected by separating the filter from the bottom of the cylinder. Filter and sediment then were processed for histology cell block. Remarkable conservation of tissue architecture was noted with this technique, with results comparable to findings in endocervical curettage. RESULTS: Seventy specimens with suspected AGUS were processed as described above, 57 of which contained interpretable tissue materials. Of these 57 specimens, 38 (67%) were found to be abnormal and 19 represented benign diagnoses. Final diagnoses in the abnormal AGUS group included 6 cases with AGUS that could not be classified further, 2 cases with invasive squamous cell carcinoma, 15 cases with high grade squamous intraepithelial lesions, 14 cases with adenocarcinoma (in situ or invasive) (including 3 with endometrial adenocarcinoma), and 1 case with endometrial hyperplasia. CONCLUSIONS: Inverted filter sedimentation is a useful technique in the differential diagnosis of AGUS. It can provide a training tool with which to increase the sensitivity and specificity of AGUS diagnoses as well as occasionally provide immediate tissue confirmation. PMID- 11038423 TI - Can CD34 discriminate between benign and malignant hepatocytic lesions in fine needle aspirates and thin core biopsies? AB - BACKGROUND: Distinguishing well differentiated hepatocellular carcinoma (HCC) from benign hepatocellular lesions is a well recognized problem in fine-needle aspiration (FNA) cytology. The endothelial cell marker CD34 is negative in normal hepatic sinusoids and stains vessels diffusely in HCC. This feature is useful in distinguishing benign from malignant hepatocytic lesions in histological specimens, although benign lesions may show focal positivity for CD34 confined to periportal and periseptal areas. In this study, we assess the role of CD34 in cell block and thin core biopsy material from benign and malignant hepatocellular lesions, and compare it with reticulin staining. METHODS: Cell blocks and thin core biopsies were assessed from 40 cases of HCC and 25 benign hepatocytic lesions. HCCs were scored for nuclear grade. Sections were stained for CD34 antigen and scored semi-quantitatively. Previously performed reticulin stains were reviewed. RESULTS: Thirty three of 40 HCCs (82.5%) showed diffuse positivity for CD34. The other seven cases showed either focal positivity (four cases), minimal positivity (two cases) or negative staining (one case). These results did not correlate with the nuclear grade of the tumor. Two of 25 benign cases (8%) showed diffuse positivity for CD34, 8 showed focal positivity, 11 showed minimal positivity, and 4 showed negative staining. All HCCs showed an abnormal reticulin pattern characterized by expanded trabeculae and islands, or sheets, with decreased or absent reticulin. All of the benign hepatocellular lesions showed a normal trabecular reticulin pattern. CONCLUSIONS: Diffusely positive CD34 staining is useful to support a diagnosis of well differentiated HCC, but in our study the reticulin stain distinguished more consistently between benign and malignant lesions. PMID- 11038424 TI - Small cell carcinoma versus other lung malignancies: diagnosis by fine-needle aspiration cytology. AB - BACKGROUND: When a diagnosis of small cell carcinoma is reached in a patient with a lung mass, a surgical treatment approach is no longer considered and chemotherapy becomes the treatment of choice. The aim of this study is to compare the diagnostic accuracy of fine-needle aspiration cytology in the diagnosis of small cell carcinoma with the diagnoses of other lung malignancies. The capacity of this technique to distinguish between these two categories is assessed. METHODS: Two hundred fifty-nine consecutive transthoracic fine needle aspirations of lung tissue from 235 patients with histologic diagnosis of malignancy were reviewed. The aspirates were performed over a 10-year period at the University of Miami/Jackson Memorial Medical Center, Miami, Florida. Two hundred and forty-two fine-needle aspirations from 221 patients yielded satisfactory smears and were included in the study. Fourteen patients were excluded. The cytologic diagnoses were classified into 5 categories: 1) small cell carcinoma (18 smears, 7%); 2) other lung malignancies (158 smears, 65%); 3) suspicious for malignancy (19 smears, 8%); 4) inflammatory process (7 smears, 3%); and 5) negative for malignancy (40 smears, 17%). RESULTS: The histologic diagnoses were divided into two groups: small cell carcinomas (29 smears, 12%), and other lung malignancies (213 smears, 88%). The efficiency of fine-needle aspiration cytology in the diagnosis of these two groups was 96% versus 88%, respectively, with an equal specificity of 100%, and a sensitivity of 67% versus 81%. Once the diagnosis of malignancy was established, fine-needle aspiration cytology was found to be highly accurate in distinguishing small cell carcinoma from other neoplasms. CONCLUSION: We conclude that fine-needle aspiration cytology of the lung is an accurate diagnostic tool for the diagnosis of lung malignancies and is an excellent technique for distinguishing small cell carcinoma from other malignant neoplasms. It can be used with confidence to select treatment modalities and to avoid unnecessary surgeries in patients with lung malignancies. PMID- 11038425 TI - Fine-needle aspiration cytology of breast carcinoma with endocrine differentiation. AB - BACKGROUND: The value of fine-needle aspiration (FNA) in atypical proliferative to in situ to low grade invasive breast lesions remains limited due to the overlapping cytologic features of these entities. In the current study the authors review the FNA cytology of endocrine carcinoma and identify common cytologic features that allow for the diagnosis of this uncommon, low grade subtype of mammary malignancy. METHODS: The histopathology files from the medical practices of both authors were searched between January 1996 and May 1999 and yielded six cases of endocrine carcinoma. The clinical history and all previous FNA smears were reviewed. RESULTS: All six patients were elderly women (mean age of 72 years). Four patients presented with breast masses, one patient presented with nipple discharge, and one patient presented with both a breast mass and nipple discharge. All six surgical specimens showed endocrine ductal carcinoma in situ (E-DCIS), with four specimens showing invasive endocrine carcinoma, two of which were labeled as mucinous carcinoma. All invasive components showed the same histomorphologic and immunohistochemical profiles as the in situ components. Cytology demonstrated common features of the cellular smears with clusters and single, monomorphic plasmacytoid tumor cells that possessed moderate amounts of eosinophilic, granular cytoplasm and eccentric nuclei with fine chromatin and inconspicuous nucleoli. In four cases, additional fragments of fine and elaborate papillary fronds also were present. CONCLUSIONS: The cytologic smear diagnosis of endocrine carcinoma is assisted by the presence of plasmacytoid tumor cells and arborizing papillary fronds. PMID- 11038426 TI - Soft tissue aspiration cytopathology. AB - BACKGROUND: Fine-needle aspiration (FNA) biopsy as a diagnostic modality for the pathologic evaluation of soft tissue neoplasms and non-neoplastic soft tissue mass lesions is uncommon and controversial. This procedure contrasts with more traditional diagnostic methods such as marginal excision, incisional (open) biopsy, or even core biopsy to procure tissue from somatic sites. METHODS: The authors reviewed the results of cytopathologic diagnoses obtained by fine-needle aspiration biopsy over a consecutive 11-month period in patients that presented primarily with a palpable soft tissue mass. A few patients with deep non-palpable soft tissue masses also were evaluated by radiologically guided FNA. Cytopathologic diagnoses were verified by different means including tissue examination either by concurrent cell block or subsequent surgical biopsy, flow cytometry, clinical outcome, or repetition of the FNA procedure. Patients were followed for a minimum of one year to evaluate the mass clinically, to determine whether any further therapy was administered, and to assess disease status. RESULTS: Eighty-two aspirates were performed without complications from 77 patients ranging from 12-88 years of age (mean = 50 yrs.) with men outnumbering women 1.5:1. Soft tissue masses were most common in the extremities (41 cases), followed by the trunk (34 cases), retroperitoneum (5 cases), and head and neck (2 cases). Fine-needle aspirates were diagnosed as malignant in 42 (51%), benign in 32 (39%), nondiagnostic in 6 (7%), and atypical in 2 (2%) cases. Malignant aspirates were comprised of 24 sarcomas (57%), 9 carcinomas (21%), 6 malignant lymphomas (14%), and 3 melanomas (7%). Twenty-two aspirates (52%) had an initial diagnosis of malignancy, whereas 18 (43%) represented metastatic and 2 (5%) recurrent neoplasms. Confirmation of the cytopathologic diagnosis was by concurrent or subsequent tissue examination in 57%, flow cytometry in 5%, clinical outcome in 34%, and repeat aspiration in 4%. One false negative and no false positive diagnoses were issued for a sensitivity and specificity of 100% and 97% respectively in distinguishing benign and malignant lesions by FNA. Of the malignant aspirates, 83% could be subtyped whereas 72% of benign aspirates were correctly subtyped. For primary soft tissue sarcomas, 12 of 19 (63%) were accurately subtyped. In 48% of cases a concurrent cell block was obtained and found diagnostically useful in 54% of them. CONCLUSIONS: Aspiration cytopathology of soft tissue mass lesions using FNA biopsy can be an accurate and minimally invasive method for the initial pathologic diagnosis of primary benign and malignant soft tissue masses, for the pathologic confirmation of metastatic tumors to soft tissue, and for the documentation of locally recurrent soft tissue neoplasms. FNA cytopathology is capable of specifically subtyping a large percentage of primary and metastatic soft tissue tumors if cellular material either in the form of a cell block or flow cytometry is obtained in addition to cell smears. PMID- 11038428 TI - Fine-needle aspiration biopsy of papillary thyroid carcinoma: diagnostic utility of cytokeratin 19 immunostaining. AB - BACKGROUND: Papillary thyroid carcinoma is the most common malignant neoplasm of the thyroid gland, and fine-needle aspiration biopsy (FNAB) often is the initial diagnostic method used in its detection. Prior studies have shown that immunohistochemical staining for various cytokeratins in general, and cytokeratin 19 (CK19) in particular, can be applied as an ancillary technique for diagnosing papillary thyroid carcinoma in histologic specimens. In the current study the authors assessed the diagnostic utility of CK19 to detect papillary carcinoma effectively in cytologic preparations of thyroid FNABs. METHODS: Immunocytochemical staining with CK19 was performed on cytologic aspirates from 37 papillary thyroid carcinomas and 36 other lesions of the thyroid (14 follicular adenomas, 10 multinodular goiters, 5 cases of Hashimoto thyroiditis, 6 oncocytic [Hurthle cell] neoplasms, and 1 follicular carcinoma). All cases included in the study had a corresponding histopathology specimen. RESULTS: Positive immunocytochemical reactivity for CK19 was identified in 34 of 37 papillary carcinomas and in 1 of 36 other thyroid lesions (sensitivity of 92% and specificity of 97%). Although the strongest reactivity was obtained in methanol fixed thin layer preparations, the antibody also was effective in detecting papillary carcinoma in alcohol fixed and air-dried smears. The single false positive case was a follicular adenoma with focal areas of papillary hyperplasia. All other aspirates including those from cases of Hashimoto thyroiditis, multinodular goiter, follicular adenoma, oncocytic neoplasms, and follicular carcinoma were negative. CONCLUSIONS: CK19 is an effective, highly sensitive, and specific ancillary tool for the diagnosis of papillary carcinoma in thyroid FNABs. PMID- 11038427 TI - Use of E-cadherin and CD44 aids in the differentiation between reactive mesothelial cells and carcinoma cells in pelvic washings. AB - BACKGROUND: The presence of malignant cells in peritoneal washings is an independent prognostic factor in the evaluation of gynecologic malignancies. The differentiation between reactive mesothelial cells and carcinoma cells can be a diagnostic challenge based on morphology alone. The expression of some cell adhesion molecules may be helpful in the differential diagnosis. METHODS: To evaluate the specificity of 2 transmembrane cell adhesion proteins (E-cadherin and CD44) in the differentiation of mesothelial cells from carcinoma cells in pelvic washings, formalin fixed, paraffin embedded cell blocks of pelvic washings from 19 cases of metastatic ovarian adenocarcinoma and 16 cases of benign peritoneal washings were immunostained with monoclonal antibodies to E-cadherin and CD44. The staining patterns were evaluated blindly by three observers. Positive staining was defined as uniform membranous staining for each marker. RESULTS: Fourteen benign peritoneal washings (87.5%) demonstrated immunoreactivity with anti-CD44. On the contrary, only four adenocarcinomas (21.1%) demonstrated anti-CD44 immunoreactivity. E-cadherin expression was identified in only 2 benign peritoneal washings (12.5%) whereas 16 adenocarcinomas (84.2%) strongly expressed E-cadherin. The differences in immunostaining for both CD44 and E-cadherin between benign and malignant peritoneal washings were statistically significant. The combination of positive staining for E-cadherin and negative staining for CD44 was 100% specific for metastatic adenocarcinoma, whereas a combination of negative staining for E cadherin and positive staining for CD44 was 100% specific for reactive mesothelial cells. CONCLUSIONS: Both E-cadherin and CD44 reliably distinguish reactive mesothelial cells from adenocarcinoma. The combination of E cadherin/CD44 is highly specific and is a useful diagnostic adjunct with which to distinguish benign reactive mesothelial cells from adenocarcinoma in pelvic washings. PMID- 11038429 TI - Diagnostic utility of calretinin immunohistochemistry in cytologic cell block preparations. AB - BACKGROUND: Calretinin (CR) is a valuable marker in the immunohistochemical distinction between malignant mesothelioma (MM) and adenocarcinoma (ACA) in tissue sections. However, there is limited and conflicting data regarding the utility of CR in this differential diagnosis on cytologic material, especially cell block preparations. Also, the possible role of CR in the distinction of papillary serous borderline tumor (SBT) cells from reactive mesothelial cells in peritoneal washings has not been examined. METHODS: Formalin-fixed paraffin embedded cell block specimens of cytologic fluids, washings, and aspirates with a suspicious or positive cytologic diagnosis and a confirmed diagnosis of MM (29 cases), ACA (39 cases), and SBT (10 cases) were used for CR immunohistochemistry (IHC). RESULTS: Moderate to strong staining in > 50% of tumor cells was noted in 26 of 29 (90%) MMs but in only 3 of 39 (8%) ACAs and 1 of 10 (10%) SBTs. Admixed reactive mesothelial cells (when present) were strongly positive in all fluid specimens, but the staining pattern of benign non-reactive mesothelial cells in washing specimens was less reliable. CONCLUSIONS: CR is both a sensitive and specific marker of reactive and neoplastic mesothelial cells in cytologic cell block preparations and is thus useful in the differential diagnosis of MM and metastatic ACA in malignant effusions. However, CR IHC does not appear to allow definitive identification of SBT in peritoneal washings. PMID- 11038431 TI - Author reply PMID- 11038430 TI - Diagnosis of lymphoma by fine-needle aspiration cytology using the revised European-American classification of lymphoid neoplasms. PMID- 11038432 TI - Carbamazepine level-A in vivo-in vitro correlation (IVIVC): a scaled convolution based predictive approach. AB - A method is presented for prediction of the systemic drug concentration profile from in vitro release/dissolution data for a drug formulation. The method is demonstrated using four different tablet formulations containing 200 mg carbamazepine (CZM), each administered in a four way cross-over manner to 20 human subjects, with 15 blood samples drawn to determine the resulting concentration profile. Amount versus time dissolution data were obtained by a 75 rpm paddle method for each formulation. Differentiation, with respect to time, of a monotonic quadratic spline fitted to the dissolution data provided the dissolution rate curve. The dissolution curve was through time and magnitude scaling mapped into a drug concentration curve via a convolution by a single exponential, and the estimated unit impulse response function. The method was tested by cross-validation, where the in vivo concentration profiles for each formulation were predicted based on correlation parameters determined from in vivo-in vitro data from the remaining three formulations. The mean prediction error (MPE), defined as the mean value of 100% x(observed-predicted)/observed was calculated for all 240 cross-validation predictions. The mean values of MPE were in the range of 10-36% (average 22%) with standard deviations (S.D.s) in the range of 9-33% (average 13%), indicating a good prediction performance of the proposed in vivo-in vitro correlation (IVIVC) method. PMID- 11038433 TI - Non-linear regression analysis with errors in both variables: estimation of co operative binding parameters. AB - Four different parameter estimation criteria, the geometric mean functional relationship (GMFR), the maximum likelihood (ML), the perpendicular least-squares (PLS) and the non-linear weighted least squares (WLS), were used to fit a model to the observed data when both regression variables were subject to error. Performances of these criteria were evaluated by fitting the co-operative drug protein binding Hill model on simulated data containing errors in both variables. Six types of data were simulated with known variances. Comparison of the criteria was done by evaluating the bias, the relative standard deviation (S.D.) and the root-mean-squared error (RMSE), between estimated and true parameter values. Results show that (1) for data with correlated errors, all criteria perform poorly; in particular, the GMFR and ML criteria. For data with uncorrelated errors, all criteria perform equally well with regard to the RMSE. (2) Use of GMFR and ML lead to lower values for S.D. but higher biases compared with WLS and PLS. (3) WLS performs less well when equal dispersion is applied to the two observed variables. PMID- 11038434 TI - Bioavailability of melatonin in humans after day-time administration of D(7) melatonin. AB - Absolute bioavailability of the neurohormone melatonin (MLT) was studied in 12 young healthy volunteers (six males, six females) after administration at midday, on two separate occasions, of 23 microg by intravenous (i.v.) infusion and 250 microg by oral solution of D(7) MLT, a molecule in which seven deuterium atoms replace seven hydrogen atoms. Exogenous (D(7)) and endogenous (D(0)) MLT were quantified simultaneously but separately by a highly specific assay: gas chromatography/negative ion chemical ionization mass spectrometry, developed in our laboratory, which enabled us to go down to 0.5 pg/mL in plasma samples. After i.v. administration, the maximum plasma concentration (C(max)) and the area under the plasma concentration-time curve (AUC) values were significantly different in male and female subjects, but there was no significant gender difference in total body clearance normalized to body weight: 1.27+/-0.20 L/h/kg and 1.18+/-0.22 L/h/kg for males and females, respectively. The apparent terminal half-life (t(1/2(z))) values were 36+/-2 and 41+/-10 min, respectively. After oral administration, pharmacokinetic parameters used to quantify bioavailability were near three-fold greater in female subjects than in males, with large inter individual variations. The maximum plasma MLT concentration C(max)+/-S.D. was found at 243.7+/-124.6 pg/mL and 623.6+/-575.1 pg/mL for male and female subjects respectively, while the mean values for AUCs were 236+/-107 pg.h/mL and 701+/-645 pg.h/mL. The absolute bioavailability of MLT was from 1 to 37%: mean=8.6+/-3.9% and 16.8+/-12.7% for male and female subjects, respectively. PMID- 11038435 TI - Effect of food on the pharmacokinetics of osmotic controlled-release methylphenidate HCl in healthy subjects. AB - The effect of a high-fat meal on the pharmacokinetics of OROS(R) (methylphenidate HCl), an osmotic controlled-release formulation of methylphenidate HCl, was investigated in healthy subjects. Mean peak methylphenidate plasma concentrations occurred slightly later and peak methylphenidate plasma concentrations and AUC values were slightly higher (approximately 10-30%) in the presence of food than in the absence of food. There was no difference in the plasma elimination half life of methylphenidate between the fed and fasted state. Similarly, peak concentrations and AUC values for alpha-phenyl-2-piperidine acetic acid (PPA; ritalinic acid), the major metabolite of methylphenidate, were slightly higher in the presence of food than in its absence. The 90% confidence interval (CI) for the treatment ratio (fed/fasted) for C(max) was 120.6%-140. 0% for the 18-mg dose and 105.4-119.3% for a 36-mg dose. The 90% CI for the treatment ratio for AUC(infinity) was 114.6-125.7% for the 18-mg dose and 115.1-124.6% for the 36-mg dose. PPA levels were measured following the 18-mg dose and the 90% CI of the treatment ratio was 106.8-115.4% for C(max) and 97.9-103.6% for AUC(infinity). These results indicate the absence of dose dumping from OROS (methylphenidate HCl) in the presence of food. Food does not impede drug absorption and OROS (methylphenidate HCl) may be administered in the fed or fasted state. There were no differences in the adverse event profile between the fed and fasted state. PMID- 11038436 TI - Effect of a high-fat meal on thalidomide pharmacokinetics and the relative bioavailability of oral formulations in healthy men and women. AB - The effect of food on the oral pharmacokinetics of thalidomide and the relative bioavailability of two oral thalidomide formulations were determined in an open label, single dose, randomized, three-way crossover study. Five male and eight female healthy volunteers received a single oral dose of 200 mg Celgene thalidomide capsules under fasted and non-fasted conditions, and a single dose of 200 mg tablets of Serral thalidomide under fasted conditions. The high-fat meal resulted in a 0.5-1.5 h absorption lag time, an increased mean C(max), a decreased mean AUC and a delay in mean T(max). The Serral tablet formulation resulted in a lower mean C(max), and slower terminal decline in plasma thalidomide concentrations compared with both Celgene treatments. Mean C(max) concentrations were 1.99+/-0.41 microg/mL (range 1.28-2.76) within 4.00+/-1.13 h (2-5) for the Celgene formulation fasted, 2.17+/-0.51 microg/mL (1.43-3.01) within 6.08+/-2.33 h (3-12) for the Celgene formulation with food, and 1. 05+/ 0.31 microg/mL (0.62-1.65) within 6.23+/-1.88 h (5-10) for the Serral formulation fasted. Mean terminal half-lives were 13.50+/-6. 77 h for the Serral product, compared with 5.80+/-1.72 h and 5. 09+/-1.03 h for Celgene fasted and fed, respectively. Celgene's formulation exhibited slightly greater bioavailability than Serral's formulation, with mean ratios of 122% and 110% for Ln-transformed AUC(0-t) and AUC(0-infinity), respectively. The mean C(max) for the Celgene formulation was approximately two times greater than Serral's. Food delayed the onset of absorption of by 0.5-1.5 h, but had little effect on the extent of absorption from the Celgene capsule. Under fasted conditions, the Celgene thalidomide resulted in a two-fold greater C(max) and 10% greater AUC(0-infinity) than the Serral formulation. PMID- 11038437 TI - Japanese family with an autosomal dominant chromosome instability syndrome: a new neurodegenerative disease? AB - We report on a Japanese family having an autosomal dominant neurodegenerative disease with chromosomal instability and radiosensitivity. Clinical manifestations of affected members included short stature, osteoporosis, severe dental caries, and various neurological abnormalities, such as mental retardation, depression, dysarthria, hyperreflexia, and ataxic gait. MRI demonstrated a markedly atrophic spinal cord and degeneration of the white matter. Cytogenetic examination showed spontaneous chromosome rearrangements at 14q11.2 and hypersensitivity to radiation and bleomycin. The degree of these cytogenetic abnormalities was significantly higher in the patients than in normal controls but lower than in patients with ataxia telangiectasia or Nijmegen breakage syndrome. Moreover, genetic anticipation was observed in this family: the age of disease onset became earlier, MRI abnormalities more extensive, and the chromosome hypersensitivity to radiation increased in successive generations. We speculate that a basic defect in this family is a mutation in the gene that is responsible for DNA double-strand breakage repair. PMID- 11038438 TI - Familial cryptic translocation with del 4q34-->qter and dup 12pter-->p13 in sibs with tracheal stenosis: clinical, classical and molecular cytogenetic studies and CGH analyses from archival placental tissues evidencing tertiary trisomy 4 in one abortion specimen. AB - We report on two retarded half-sibs of different sex and seemingly normal karyotype who had the same syndrome of minor anomalies, heart defect and a distal tracheal stenosis, and who shared a healthy mother. These findings raised suspicions of a cryptic chromosome translocation. A translocation t(4;12)(q34;p13), balanced in the mother and unbalanced in the sibs with loss of terminal 4q and gain of terminal 12p regions, was verified by FISH using whole chromosome painting, subtelomeric and YAC probes. Clinical features could be explained by partial monosomy 4q and partial trisomy 12p. Tracheal stenosis was interpreted as a consequence of the same developmental disturbance leading to esophageal atresia and tracheo-esophageal fistula. It was attributed to the 4q deletion in which esophageal atresia as also respiratory difficulties and airway obstructions had been described. Paraffin-embedded placental tissues were available from three of the five abortions of the mother allowing DNA extraction and comparative genome hybridization (CGH). Two of the abortion specimens had the same der(4)t(4;12)(q34;p13) unbalanced translocation as identified in the sibs. In the third abortion specimen, suspicious of triploidy because of partial hydatidiform mole, CGH uncovered a tertiary trisomy 4 resulting from a 3:1 segregation of the translocation chromosomes and their homologs during maternal meiosis I. Differences in CGH results using DNA generated directly or after DOP PCR were explained by DNA fragmentation in paraffin-embedded tissues and unequal amplification. Am. J. Med. Genet. 94:271-280, 2000. PMID- 11038439 TI - Ablepharon-macrostomia syndrome: first report of familial occurrence. AB - Ablepharon-macrostomia syndrome (AMS) is a rare condition comprising severe deficiency of the anterior lamella of both eyelids, abnormal ears, macrostomia, anomalous genitalia, redundant skin, and absence of lanugo. There is no agreement about cause; some authors suggest autosomal recessive inheritance. We describe familial occurrence of AMS in a girl, sister of a previously reported patient. The father has facial anomalies that suggest autosomal dominant inheritance. Am. J. Med. Genet. 94:281-283, 2000. PMID- 11038440 TI - Maternal uniparental disomy of chromosome 16 and body stalk anomaly. AB - We report on a fetus with placental trisomy 16, maternal uniparental disomy (UPD), and body stalk anomaly. Body stalk anomaly is a rare, fatal developmental abnormality consisting of a defective abdominal wall with abdominal organs in a sac outside the abdominal cavity covered by amnion adherent to the placenta with absence or severe shortness of the umbilical cord. Trisomy 16 was identified in the placenta in all cells. Amniocentesis was karyotypically normal. Parental origin studies showed maternal UPD for chromosome 16 in post-termination fetal tissue. The cause of the body stalk anomaly is not clearly defined. There are no other reports of placental karyotype or UPD investigations with body stalk anomaly. To our knowledge, this is the first report of placental trisomy 16, UPD in fetus, and body stalk anomaly, suggesting placental insufficiency or imprinting effects as cause of this anomaly. Am. J. Med. Genet. 94:284-286, 2000. PMID- 11038441 TI - Spectrum of Schwartz-Jampel syndrome includes micromelic chondrodysplasia, kyphomelic dysplasia, and Burton disease. AB - Follow-up and re-evaluation of four patients originally described as examples of severe infantile "micromelic chondrodysplasia" resembling Kniest disease, "kyphomelic dysplasia," and "Burton skeletal dysplasia" revealed the diagnosis of Schwartz-Jampel syndrome (SJS, myotonic chondrodysplasia) in all of them. SJS may be suspected in neonates with Kniest-like chondrodysplasia, congenital bowing of shortened femora and tibiae, and facial manifestations consisting of a small mouth, micrognathia, and possibly pursed lips. The disorder must be differentiated from the Stuve-Wiedemann syndrome, a genetically distinct myotonic chondrodysplasia with similar clinical but different skeletal changes and an unfavorable early prognosis. The demise of "kyphomelic dysplasia" as a nosological entity reemphasizes the symptomatic nature of congenital bowing of the long bones. PMID- 11038442 TI - Mild phenotype due to tandem duplication of l7p11.2. AB - We present a mildly affected girl with de novo dup(17)(p11.2p11.2). The patient was evaluated because of minor anomalies noted during a hospitalization for nonrecurrent tonic-clonic seizures associated with transient hypoglycemia. She also had unilateral renal hypoplasia and relative short stature, but at 2 years of age, she scored within the low normal range on neurodevelopmental examinations. Compared with other similar duplications, this patient represents the milder range of the spectrum for this karyotypic abnormality. Am. J. Med. Genet. 94:296-299, 2000. PMID- 11038443 TI - Novel and recurrent EBP mutations in X-linked dominant chondrodysplasia punctata. AB - Chondrodysplasia punctata (CDP) is a heterogeneous group of skeletal dysplasias characterized by stippled epiphyses. A subtype of CDP, X-linked dominant chondrodysplasia punctata (CDPX2), known also as Conradi-Hunermann-Happle syndrome, is a rare skeletal dysplasia characterized by short stature, craniofacial defects, cataracts, ichthyosis, coarse hair, and alopecia. The cause of CDPX2 was unknown until recent identification of mutations in the gene encoding Delta(8),Delta(7) sterol isomerase emopamil-binding protein (EBP). Twelve different EBP mutations have been reported in 14 patients with CDPX2 or unclassified CDP, but with no evidence of correlation between phenotype and nature of the mutation. To characterize additional mutations and investigate possible phenotype-genotype correlation, we sequenced the entire EBP gene in 8 Japanese individuals with CDP; 5 of them presented with a CDPX2 phenotypes. We found EBP mutations in all 5 CDPX2 individuals, but none in non-CDPX2 individuals. Three of these CDPX2 individuals carried novel nonsense mutations in EBPand the other two, separate missense mutations that had been reported also in different ethnic groups. Our results, combined with previous information, suggest all EBP mutations that produce truncated proteins result in typical CDPX2, whereas the phenotypes resulted from missense mutations are not always typical for CDPX2. Patients with nonsense mutations showed abnormal sterol profiles consistent with a defect in Delta(8), Delta(7) sterol isomerase. X-inactivation patterns of the patients showed no skewing, an observation that supports the assumption that inactivation of the EBP gene occurs at random in affected individuals. PMID- 11038444 TI - Familial dup(8)(p12p21.1): mild phenotypic effect and review of partial 8p duplications. AB - We describe a family with direct transmission of a duplication of 8p12-->8p21.1. The phenotype of affected relatives included mild mental retardation but no minor anomalies. The duplication was identified by means of GTG-banding and fluorescence in situ hybridization with a probe specific for 8p12 generated by microdissection and degenerate oligonucleotide primed-polymerase chain reaction. Assay of glutathione reductase, which has been localised to 8p21.1, was significantly increased when compared with controls with normal chromosomal constitution. To the best of our knowledge, a proximal direct duplication of 8p restricted to subbands p12-->p21.1 has not been reported so far. The reported aberration is compared with other partial duplications of 8p, in particular to inversion duplications 8p and to small direct distal duplications involving 8p23.1. Am. J. Med. Genet. 94:306-310, 2000. PMID- 11038445 TI - Trisomy of 3pter in a patient with apparent C (trigonocephaly) syndrome. AB - The C syndrome is a multiple congenital anomaly/mental retardation (MCA/MR) syndrome first described in sibs. The inheritance has been assumed to be autosomal recessive. Several authors have commented that the combination of anomalies found in the conditions suggest an underlying chromosomal anomaly and in a few apparent cases chromosome anomalies have been described. Our patient had findings consistent with the C syndrome and a duplication of 3p by use of subtelomere probes. This shows that new cytogenetic techniques continue to be important in defining the underlying cause of MCA/MR conditions. PMID- 11038446 TI - Parents' responses to disclosure of genetic test results of their children. AB - The psychological reactions of 22 parental couples and 3 single parents were investigated after disclosure of genetic test results of their children. The children were tested for the early-onset, monogenetic cancer disorder multiple endocrine neoplasia type 2. Participants came from 13 different families and were aged between 28 and 47 years. Parents who were informed that their child was a gene carrier reacted with resignation, showed moderate to high levels of test related and general anxiety, but few psychological complaints. Daily activities were disturbed in 43% of the parents with carrier-children. There was little disruption of the parents' future perspective, apart from some socioeconomic disadvantages and increased parental concern for the carrier-children. Most parents with carrier-children showed restraint with respect to short-term prophylactic treatment. Parents with favorable test results showed significantly less anxiety and no disturbance in their daily activities. They did not, however, seem to be reassured by the DNA test result. These parents questioned the reliability of the DNA test, wanted confirmation of the test results, and were eager to continue screening of their noncarrier children. Parents, especially those with a lower level of education and/or a pessimistic view of the future, were distressed by unfavorable test results. Additional counseling is advised to prevent parents of carrier-children worrying unnecessarily, or parents with children in whom the disease gene was not found being not reassured. Am. J. Med. Genet. 94:316-323, 2000. PMID- 11038447 TI - Quantitative genetic analysis of circulating levels of biochemical markers of bone formation. AB - Carboxyterminal propeptide of type 1 collagen (PICP) and bone Gla-protein osteocalcin (BGP) are the most important components of the organic bone matrix and play a key role in bone formation. To investigate whether and to what extent variation of the plasma levels of these indices of bone turnover depends on genetic factors, we studied 355 adults belonging to nuclear pedigrees. Genetic analysis was carried out in 2 steps: 1) variance decomposition analysis was performed using the FISHER statistical package; and 2) complex segregation analysis implemented in the program package MAN. The effect of age and gender differences, gender hormones, as well as PTH and vitamin-D (calcidiol) plasma levels were evaluated simultaneously with the parameters of variance analysis. The results showed that about 50% of PICP variation is attributable to genetic factors. The effect of age was significant among men and postmenopausal women, whereas calcidiol influenced variation of PICP in premenopausal women. The results of variance analysis showed that some 40% of BGP, adjusted for confounding variables, can be explained in genetic factors. Age and PTH were important covariates for osteocalcin in men and premenopausal women. Exploration of the maximum likelihood estimates of the various hypotheses concerning the mode of intergenerational transmission of PICP and BGP demonstrated a good correspondence to the Mendelian mode of inheritance (i.e., major gene effect). PMID- 11038448 TI - Newborn infant with lethargy, poor feeding, dehydration, hypothermia, hyperammonemia, neutropenia, and thrombocytopenia. PMID- 11038450 TI - Behold the CHILD, it's only one: CHILD syndrome is not caused by deficiency of 3 beta-hydroxysteroid-Delta 8, Delta 7-isomerase. PMID- 11038449 TI - Ectodermal abnormalities associated with methimazole intrauterine exposure. PMID- 11038452 TI - Combined first trimester nuchal translucency and second trimester biochemical screening tests among normal pregnancies. AB - We prospectively examined whether first trimester nuchal translucency (NT) and second trimester triple test (TT) results are correlated, and determined overlapping and mutual screen-positive rates. Results of NT, TT, amniocentesis and pregnancy outcome were obtained in 508 normal pregnancies. Inter-test correlation was performed by comparing the likelihood ratios (LR). Overlapping of screen-positive cases, of NT and TT, was determined by comparing mutual risks for Down syndrome (DS) livebirth of > or = 1:380. Combined screen-positive rates were evaluated by using summation risk (NT and/or TT exhibiting a risk > or = 1:380) and calculated risk (new risk > or / =1:380, based on multiplication of LR(NT) and LR(TT)). Screen-positive rates between NT and TT differed significantly and when either test showed an increased risk for DS, the probability of the other to predict the same was negligible (p<0.001). Overall screen-positive rates, at a risk > or = 1:380, were 2% and 5.7% for NT and TT, respectively. Summation and calculated combining methods were associated with 7.5% and 2.0% screen-positive rates, respectively. Amniocentesis was performed on 20.7% of the cases, mostly screen-negative ones. Our results showed that, in normal pregnancies, NT and TT do not correlate and that their combined calculated risk in normal pregnancies is associated with a low screen-positive rate of 2.0%. PMID- 11038454 TI - Response to hallahan et al PMID- 11038453 TI - First trimester biochemical screening for Down syndrome: free beta hCG versus intact hCG. AB - To compare free beta hCG versus intact hCG in first trimester Down syndrome screening we analysed 63 cases of Down syndrome and 400 unaffected control pregnancies between 10 and 13 weeks' gestation. The Down syndrome median multiple of the median (MoM) was significantly higher (p=0.001) for free beta hCG (1.89 MoM) than for intact hCG (1.37 MoM). Although distributions for free beta hCG (unaffected, 0.2157; DS, 0.2322) are wider than for intact hCG (unaffected, 0.1697; DS, 0.2158), overall 27% of Down syndrome cases were above the 95th percentile for free beta hCG compared to 19% for intact hCG. Combined with maternal age, free beta hCG detected 45% of Down syndrome pregnancies at a 5% false positive rate. Intact hCG combined with maternal age demonstrated a detection efficiency comparable to maternal age alone (35% versus 32%). In contrast, a recent study (Haddow et al., 1998-NEJM 338: 955-961) indicated that intact hCG yielded a higher first trimester Down syndrome detection efficiency than free beta hCG (29% versus 25% respectively). Re-analysis of distribution parameters in the Haddow et al. study, however, show that free beta hCG was actually the better marker (23% detection for intact hCG versus 29% for free beta hCG). PMID- 11038455 TI - The influence of parity and gravidity on first trimester markers of chromosomal abnormality. AB - We have studied changes in first trimester fetal nuchal translucency (NT) and maternal serum free beta-hCG and PAPP-A with gravidity and parity in 3252 singleton pregnancies unaffected by chromosomal abnormality or major pregnancy complications. We have shown that gravidity and parity is associated with a small but progressive decrease in fetal NT and a small but progressive increase in free beta-hCG and PAPP-A. None of these small changes with increasing gravidity or parity are statistically significant and hence correction for these variables is not necessary when considering first trimester screening for chromosomal abnormalities. PMID- 11038456 TI - Fetal DNA in maternal plasma is elevated in pregnancies with aneuploid fetuses. AB - Current non-invasive screening methods for the prenatal diagnosis of fetal aneuploidies are hampered by low sensitivities and high false positive rates. Attempts to redress this situation include the enrichment of fetal cells from maternal blood, or the use of fetal DNA in the plasma of pregnant women. By the use of real-time quantitative polymerase chain reaction (PCR) it has recently been shown that circulatory male fetal DNA in maternal plasma is elevated in pregnancies with trisomy 21 fetuses. In this independent study we confirm and extend upon these results by showing that the levels of fetal DNA are also elevated in pregnancies with other chromosomal aneuploidies (mean=185.8 genome equivalents/ml; range=62.2-471.7) when compared to pregnancies with normal male fetuses (mean=81.9 genome equivalents/ml; range=28.8-328.9), p=0.005. This elevation was greatest for fetuses with trisomy 21, whereas it was not significant for fetuses with trisomy 18, p=0.356. These data suggest that a quantitative analysis of such fetal DNA levels may serve as an additional marker for certain fetal chromosomal abnormalities, in particular for trisomy 21. PMID- 11038457 TI - Management of fetal thyroid goitres: a report of 11 cases in a single perinatal unit. AB - Fetal thyroid goitres may reveal hormonal imbalance. This can jeopardize neurological development and fetal outcome even when early postnatal treatment is provided. We report a series of 11 goitres diagnosed antenatally in women with past or present thyroid disorders or discovered fortuitously on ultrasound scan. Fetuses presented with hyperthyroidism in three cases and hypothyroidism in eight. Hypothyroidism was iatrogenic in five cases, due to maternal anti-thyroid drugs. Hyperthyroidism was induced by transplacental transfer of thyroid stimulating antibodies (TSHrab). Accurate diagnosis of fetal thyroid status was obtained by fetal blood sampling but this invasive method was deemed necessary only in four cases as maternal clinical and biological data and ultrasound signs provided sufficient information to infer the type of thyroid disorder in the remaining patients. Fetal therapy relied on reduction of maternal antithyroid medication and, in selected cases, intra-amniotic injection of levothyroxin in hypothyroidism, and on administration of antithyroid drugs in hyperthyroidism. All newborns were healthy and none displayed consequences of severe thyroid imbalance. No caesarean section was performed for dystocia. Fetal thyroid goitres can be managed successfully with selected use of invasive diagnostic and therapeutic techniques. PMID- 11038458 TI - A novel mutation detected by temporal temperature gradient gel electrophoresis led to the confirmative prenatal diagnosis of a Hispanic CF family. AB - Mutational analysis of 30 recurrent known mutations detects only about 58% of Hispanic cystic fibrosis (CF) chromosomes. The low mutation detection rate has greatly hindered prenatal diagnosis and carrier testing of Hispanic families who have multiple affected children with unidentified cystic fibrosis transmembrane conductance regulator (CFTR) mutations. We recently employed a temporal temperature gradient gel electrophoresis (TTGE) method to effectively scan unknown mutations in the entire CFTR gene. A novel mutation, 2105-2117 del13insAGAAA was identified in a Hispanic family heterozygous for delta F508. The discovery of the devastating mutation facilitated the prenatal diagnosis for this family who already had two severely affected children. The fetus was found to be a compound heterozygote of delta F508/2105-2117 del13insAGAAA. This case emphasizes the importance of whole gene mutational analysis in patients with a clinical diagnosis of CF, but without the identifiable DNA mutations by routine mutation analysis. Finding of CF mutations in the patient would allow proper genetic counselling and prenatal and carrier detection of at-risk family members. PMID- 11038460 TI - Prenatal diagnosis of the neuronal ceroid lipofuscinoses. PMID- 11038459 TI - Evaluation of the prenatal diagnosis of limb reduction deficiencies. EUROSCAN Study Group. AB - Ultrasound scans in the mid-trimester of pregnancy are now a routine part of antenatal care in most European countries. Using data from registries of congenital anomalies a study was undertaken in Europe. The objective of the study was to evaluate prenatal detection of limb reduction deficiencies (LRD) by routine ultrasonographic examination of the fetus. All LRDs suspected prenatally and all LRDs (including chromosome anomalies) confirmed at birth were identified from 20 Congenital Malformation Registers from the following 12 European countries: Austria, Croatia, Denmark, France, Germany, Italy, Lithuania, Spain, Switzerland, The Netherlands, UK and Ukrainia. These registries are following the same methodology. During the study period (1996-98) there were 709,030 births, and 7,758 cases with congenital malformations including LRDs. If more than one LRD was present the case was coded as complex LRD; 250 cases of LRDs with 63 (25.2%) termination of pregnancies were identified including 138 cases with isolated LRD, 112 with associated malformations, 16 with chromosomal anomalies and 38 non chromosomal recognized syndromes. The prenatal detection rate of isolated LRD was 24.6% (34 out of 138 cases) compared with 49.1% for associated malformations (55 out of 112; p<0.01). The prenatal detection of isolated terminal transverse LRD was 22.7% (22 out of 97), 50% (3 out of 6) for proximal intercalary LRD, 8.3% (1 out of 12) for longitudinal LRD and 0 for split hand/foot; for multipli-malformed children with LRD those percentages were 46.1% (30 out of 65), 66.6% (6 out of 9), 57.1% (8 out of 14) and 0 (0 out of 2), respectively. The prenatal detection rate of LRDs varied in relation with the ultrasound screening policies from 20.0% to 64.0% in countries with at least one routine fetal scan. PMID- 11038461 TI - Prenatal diagnosis of X-linked hyper-IGM syndrome by direct detection of mutation Q220X in the CD40L gene using PCR-mediated site directed mutagenesis. AB - We present the first report of prenatal diagnosis of X-linked hyper-IgM syndrome by PCR-mediated site directed mutagenesis (PSM) in a woman known to carry the Q220X mutation in the CD40L gene. Using the simple PSM assay, the Q220X mutation was identified by chorionic villous sampling (CVS) at 11 weeks' gestation and the pregnancy was terminated. PMID- 11038462 TI - Normal outcome after prenatal diagnosis of thrombosis of the torcular Herophili. AB - We report a case of prenatal diagnosis of thrombosis of the torcular Herophili. Detection at 22 weeks' gestation by ultrasound scan of an anechoic mass located immediately above the tentorium led to magnetic resonance imaging (MRI) being performed at 28 weeks which established the diagnosis of an isolated thrombosis of the torcular Herophili. MRI remained stable throughout pregnancy, and postnatal MRI confirmed the diagnosis at 2.5 months of age. The child is now 16 months old and developing normally. PMID- 11038463 TI - Early prenatal diagnosis of the ICF syndrome. AB - The ICF syndrome (immunodeficiency, (para)centromeric instability and facial abnormalities) is a rare autosomal recessive disorder with characteristic cytogenetic aberrations of chromosomes 1, 9 and 16 in lymphocytes. Previously, only one case has been diagnosed prenatally in the second trimester of pregnancy by fetal blood sampling. We report the first early prenatal exclusion of the ICF syndrome by chorionic villous sampling (CVS) and linkage analysis in a family with a previous affected child. The fetus was heterozygous for marker D20S850 closely linked to the ICF locus. The family was counselled of a probability of over 90% that the fetus would be unaffected. Postnatal chromosome analysis on peripheral blood was normal and thus confirmed the prenatal diagnosis. PMID- 11038464 TI - Genetic analysis of fetal nucleated red blood cells from CVS washings. AB - Chorionic villus sampling (CVS) is an established invasive prenatal diagnostic method for the detection of fetal chromosome aberrations. In 1-2% the karyotype result of CVS is inconclusive and follow-up confirmation will be required. To avoid another invasive procedure we examined fetal nucleated red blood cells (NRBCs) from CVS washings for genetic analysis. We analysed the washings of 20 chorionic villi samples of male fetuses. Fetal NRBCs were immunostained by an antibody against embryonic haemoglobin (HbE). FISH was performed with probes specific for the X and Y chromosome and the nucleus was counterstained with DAPI. Cells positive for the antibody, as well as for DAPI, were collected and stored by a semi-automated microscope. An operator reviewed those cells for their FISH signals. In 19 out of 20 CVS washings we found nucleated cells positive for HbE together with XY FISH signals. In none of the washings HbE positive cells with two X signals were found. Our results indicate that anti-HbE is a very specific antibody for identifying fetal NRBCs. NRBCs from CVS washings can be used as an additional fetal tissue for first trimester prenatal diagnosis. PMID- 11038465 TI - Sonographic and molecular diagnosis of thanatophoric dysplasia type I at 18 weeks of gestation. AB - Thanatophoric dysplasia is the most common type of lethal skeletal dysplasia. It can usually be diagnosed with ultrasound, but differential diagnosis with other osteochondrodysplasias is not always possible. Mutations in the fibroblast growth factor receptor 3 (FGFR3) gene have been demonstrated to cause two distinct subtypes of the disorder. We describe a case of thanatophoric dysplasia type I diagnosed at 18 weeks of gestation by ultrasonography. Genomic DNA obtained by chorionic villus sampling showed a C to G substitution at position 746 in the FGFR3 gene, resulting in a Ser249Cys substitution already known to be associated with type I disease. Implications for perinatal management are discussed. PMID- 11038466 TI - High levels of fetal erythroblasts and fetal extracellular DNA in the peripheral blood of a pregnant woman with idiopathic polyhydramnios: case report. AB - Abnormal amniotic fluid volume can be associated with increased maternal risk as well as perinatal morbidity and mortality. Polyhydramnios is often indicative of fetal, placental or maternal problems. In a large proportion of patients the aetiology of the disorder is unclear. Here we report on a case in which numerous fetal erythroblasts and large quantities of extracellular fetal DNA were found in the peripheral blood of a pregnant woman with idiopathic polyhydramnios bearing a male fetus. Following enrichment of erythroblasts by magnetic separation (MACS) and anti-CD71 antibodies, approximately 45-fold more erythroblasts were determined per ml peripheral maternal blood than in matched controls (231 versus 5). Single cell multiplex polymerase chain reaction (PCR) of individually micromanipulated erythroblasts showed that approximately 122 of these were of fetal origin. The concentration of extracellular fetal circulatory DNA in maternal plasma was determined by real-time quantitative PCR and shown to be almost double that of the control group (749.2 versus 404 fetal genome equivalents per ml maternal plasma). It can be speculated that the increased intrauterine pressure in polyhydramnios leads to an enhanced influx of fetal cells and free extracellular fetal DNA into the maternal circulation. This hypothesis will have to be tested with further cases. PMID- 11038467 TI - Incidental prenatal detection of an Xp deletion using an anonymous primer pair for fetal sexing. AB - We report on the incidental prenatal detection of an interstitial X-chromosomal deletion in a male fetus and his mother by fetal sexing with a primer pair recognizing an X-Y homologous locus (DXYS19), formerly unassigned on the X chromosome. The proband asked for prenatal diagnosis because of her elevated age and risk of Duchenne muscular dystrophy (DMD). Prior to molecular genetic testing for DMD, fetal sexing was carried out on DNA prepared from cultured amniocytes. PCR analysis revealed the expected Y-chromosomal product, but did not show the constitutive X-chromosomal fragment. The absence of the X-chromosomal fragment in the fetus and on one X chromosome of the mother was confirmed by Southern hybridization of HindIII restricted DNA with probe pJA1165 (DXYS19). DXYS19X was mapped to Xp22.3 by combining several approaches, including: (1) analysis of somatic cell hybrid lines containing different fragments of the human X chromosome; (2) Southern hybridization of a yeast artificial chromosome (YAC) filter panel provided by the Resource Center/Primary Database (RZPD); (3) FISH analysis; and (4) re-evaluation of two patients with interstitial deletions in Xp22.3. The extent of the deletion in the fetus was estimated by further markers from Xp22.3 and found to include the STS gene. Mental retardation could not be excluded since some mentally retarded patients exhibit overlapping deletions. PMID- 11038468 TI - Wolf-Hirschhorn syndrome due to a 3:1 segregation of a maternal balanced t(4;15)(p16.3;q11) translocation. AB - The Wolf-Hirschhorn syndrome (WHS) is characterized by severe pre- and postnatal growth retardation, specific pattern of dysmorphisms, and severe developmental delay. These clinical findings are the result of a deletion within the short arm of chromosome 4. Most cases occur de novo and are of paternal origin. Cases due to a balanced translocation are mostly of maternal origin and the deletion of distal 4p, including the WHS critical region, is often combined with a duplication of the other chromosomal segment involved in the rearrangement. Here, we report on a newborn female infant with WHS and pure tertiary monosomy due to a 3:1 segregation of a balanced maternal 4;15 translocation. In this context, importance of fluorescence in situ hybridization (FISH) with specific probes to determine the exact breakpoints in unbalanced chromosomal rearrangements with breakpoints localized around known microdeletion syndromes is emphasized. PMID- 11038469 TI - False-positive maternal serum screening in systemic lupus erythematosis: a case report. PMID- 11038470 TI - First trimester markers of trisomy 21 and the influence of maternal cigarette smoking status. PMID- 11038471 TI - Prenatal diagnosis of fragile X and Turner mosaicism in a 12-week fetus. PMID- 11038473 TI - Current awareness in prenatal diagnosis. PMID- 11038472 TI - Fetus with a 9q22q34 interstitial deletion and hygroma. PMID- 11038474 TI - Adjuvant psychological therapy for cancer patients: putting it on the same footing as adjunctive medical therapies. AB - A comparison is made between the models guiding the administration of adjuvant material remedies, such as chemotherapy, and 'adjunctive psychological therapy' (APT), in the treatment of cancer. It is argued that patients would benefit if APT were supplied subject to the same indications as adjuvant chemotherapy. PMID- 11038475 TI - Association of psychological vulnerability factors to post-traumatic stress symptomatology in mothers of pediatric cancer survivors. AB - The current study investigated whether individual differences in coping style, lifetime experience of traumatic events, perceived social support, and perceived social constraints were associated with symptoms of post-traumatic stress among 72 mothers of children who had successfully completed cancer treatment. Results suggested that more perceived social constraints and less perceived belonging support were associated with significantly more post-traumatic stress symptomatology, and this association was present after controlling for the effects of child age. Monitoring coping style and lifetime traumatic events were not significantly predictive of post-traumatic stress symptoms. The results of this study indicate that a sense of belonging to a social network as well as comfort expressing cancer-related thoughts and feelings to friends and family may play a key role in mothers' long-term adjustment to this extremely difficult life experience. PMID- 11038476 TI - Searching for safety signals: the experience of medical surveillance amongst men with testicular teratomas. AB - The aim of this study is to compare the experience of a group of men with Stage 1 testicular teratomas who were being managed through a surveillance programme (n=25) with a group of patients who had received chemotherapy for more advanced disease (n=22). The study employed a two-phase sequential design that combined quantitative and qualitative methods of data analysis. In the first phase, the hospital anxiety and depression scales (HADS) were used to screen for psychological morbidity. Twelve (48%) of the men assigned to the surveillance programme scored in the 'borderline' or 'clinical case' range on the anxiety subscale of the HADS, compared with six (27%) in the chemotherapy group. There was a significant negative correlation in the surveillance group between 'time since diagnosis' and an elevated anxiety subscale score on the HADS. Interviews were then conducted with 25 participants; a grounded theory approach was used to analyse the transcripts. The hypothesis that human beings are seekers of safety signals provided an explanatory model to account for the higher incidence of self reported anxiety amongst the men in the surveillance programme. PMID- 11038477 TI - Psychometric properties of the Japanese version of the Mental Adjustment to Cancer (MAC) scale. AB - This paper describes a study of the psychometric properties of the Japanese version of the mental adjustment to cancer (MAC) scale developed in England. The scale was completed by 455 Japanese cancer patients. The internal consistency was similar to that of the original version (Cronbach's alpha coefficients for the subscales ranged from 0.60 to 0.78) and the Japanese version had moderate to moderately high stability (correlation coefficients were above 0.64). Correlations between the MAC scale score and emotional states measured by the Profile of Mood States (POMS) indicated the concurrent validity. The results suggest that the Japanese version, like the original MAC scale, is a reliable and valid clinical research tool in Japan. PMID- 11038478 TI - The impact of abnormal mammograms on psychosocial outcomes and subsequent screening. AB - Few studies have examined the impact of abnormal mammograms on subsequent mammography screening and psychosocial outcomes specifically as a function of the length of time that has passed since the abnormal test result. This cross sectional report compared breast cancer screening practices and psychosocial outcomes among three groups of women. These groups were women who (1) never had an abnormal mammogram, (2) had an abnormal mammogram 2 or more years prior to the study's baseline interview, and (3) had an abnormal mammogram within 2 years prior to the study's baseline interview. Women who had an abnormal mammogram at least 2 years prior to the baseline interview expressed greater 10-year and lifetime risks of getting breast cancer than women who never had an abnormal mammogram. Women who had abnormal mammograms, independent of when they occurred, were substantially more worried about getting breast cancer than were women who never had abnormal mammograms. Women who had an abnormal mammogram within 2 years prior to the baseline interview were more likely to be on schedule for mammography, compared with women who never had an abnormal mammogram. PMID- 11038479 TI - Adjustment to perceived ovarian cancer risk. AB - Eighty-three women who perceived themselves to be at risk for ovarian cancer completed a battery of surveys. In addition to demographics, subjects were asked to complete the Brief Symptom Inventory, Multidimensional Health Locus of Control, Death Anxiety Scale, Taylor Anxiety Scale, Index of Sexual Satisfaction, Impact of Event Scale, and the Marlowe-Crowne Social Desirability Scale. Overall, the respondents were more similar to normal controls than to psychiatric outpatients. A correlation was drawn between higher levels of education and lower scores on the Brief Symptom Inventory, which measures characteristics such as somatization, obsessive compulsive behaviors, interpersonal sensitivity, and anxiety. However, those who had the highest scores on the Death Anxiety Scale were less likely to comply with the recommendation for a physical/gynecological examination. Patients who were most influenced by an external locus of control or 'powerful other' were more compliant with their physicians' recommendations for testing and examination. It is the authors' belief that individualized educational efforts and the presence of a solid support system may increase women's adherence to the recommended health care practices. PMID- 11038480 TI - Psychological impact of diagnosis and risk reduction among cancer survivors. AB - Life-threatening health events prompt psychological distress that may motivate individuals to reduce health risks. If so, interventions timed to take advantage of these 'teachable moments' could be particularly effective. To explore this association, early stage prostate and breast cancer patients were identified from a hospital-based tumor registry within 6 years of diagnosis. These patients (n=920) completed a mailed survey assessing the Horowitz impact of events scale, risk behaviors and readiness to change the behaviors. Breast cancer patients, younger patients and those reporting poor health status reported the greatest impact of the cancer diagnosis. Impact was inversely associated with time from diagnosis for prostate, but not breast cancer patients. Prostate patients who reported exercising regularly had lower impact scores than those who were not exercising (medians: 0.13 vs 0.56, respectively; p=0.02). Breast patients who were eating five or more fruits and vegetables reported lower impact scores than those who were not eating the recommended servings (0.75 vs 1.06, respectively; p=0.03). Breast patients who were non-smokers reported lower impact scores than smokers (0.88 vs 1.31, respectively; p=0. 02). Prospective studies are needed to understand the psychological impact of cancer diagnosis and how it might facilitate or impede the adoption of health promoting behaviors. PMID- 11038481 TI - Resource use in women completing treatment for breast cancer. AB - OBJECTIVE: To explore resources used by women completing treatment for breast cancer, how they learned about them, and the psychological factors that predicted their use. DESIGN: A questionnaire on resource use was administered as part of a randomized clinical trial which assessed subjects' psychosocial characteristics and tested the outcomes of a psychosocial intervention. SETTING: Women completing treatment for breast cancer were recruited from the oncology departments of three university-teaching hospitals in Montreal. A questionnaire gathered data on the resources used by the subjects, how they learned about them, and the role of the health care team in their decision-making. Emotional distress, dimensions of coping effort, a sense of control and optimism were also measured. RESULTS: Five categories of resources were explored; professional services, informal support networks, informational resources, support organizations and complementary therapies. Most women found out about the last two resources by themselves. Women who used cancer support organizations or complementary therapies scored high on the use of problem-solving coping and low on the use of escape/avoidance coping. In addition they were moderately optimistic, had a slightly lower sense of personal control and were somewhat more distressed than the non-users. The use of support organizations and complementary therapies appears to represent a thoughtful approach to dealing with the distress of cancer. The opinion of the oncologist regarding resource use was valued by nearly half of the sample. PMID- 11038482 TI - A focus group study of DES daughters: implications for health care providers. AB - A focus group study of women exposed to diethylstilbestrol (DES) in utero (DES daughters) was conducted to gain understanding about exposure to this drug from a patient perspective. Focus group participants reported that learning about their DES exposure was devastating; they experienced strains in their family relationships, emotional shock, a feeling that their health concerns were not appreciated by others and, to some degree, a sense of social isolation. Although many were aware of the need for special gynecological exams and high-risk prenatal care, they were frustrated by what they felt was a lack of reliable and clear information about the effects of DES exposure. Most expressed questions and anxiety about their health. Many found their communication with physicians about their DES exposure unsatisfying. They felt that physicians lacked information about the long-term health effects of DES exposure and as a result did not give them accurate information. Furthermore, they felt that physicians were dismissive of their concerns and often gave what they felt to be false reassurances. Consequently, the women developed an enduring distrust of the medical profession. The results of the study suggest implications for the delivery of health care to DES daughters. PMID- 11038484 TI - Kinetics and mechanism of the chemiluminescence associated with the free radical mediated oxidation of amino acids. AB - When amino acids are incubated in the presence of a free radical source [2,2' azobis(2-amidinopropane) dihydrocloride], only tyrosine (Tyr) and tryptophan (Trp) produce significant chemiluminescence. The relationship between the observed light intensity, the rate of the oxidation process and the substrate concentration is complex and can not be explained in terms of the formation of excited states in termination processes involving two peroxyl radicals (Russell's mechanism). The observed increase in light emission with the incubation time, for both Trp and Tyr, would indicate the participation of more than one reaction product as intermediates in the pathways leading to the production of excited molecules. However, the fact that after product accumulation a high proportion of the observed luminescence is quenched by Trolox addition, implies that the main chemiluminescent process must involve the interaction of product(s) and free radicals. From the effect of added Ebselen, it is proposed that hydroperoxides and peroxides, formed along the reaction path, are the intermediates whose accumulation leads to the observed increase in chemiluminescence with elapsed time. The observed time profiles and the proposed mechanism strongly resemble those associated with the oxidation of complex biological systems, suggesting that protein oxidation could be one of the main sources of chemiluminescence in biological oxidations. PMID- 11038483 TI - Effects of Helicobacter pylori in the stomach on neutrophil chemiluminescence in patients with gastric cancer. AB - Hypochlorous acid, which is one of the reactive oxgen species (ROS) produced from human neutrophils, is converted to cytotoxic NH(2)Cl after reaction with ammonia produced by urease in Helicobacter pylori (HP), increasing gastric mucosal injury and with potential development to gastric cancer. We compared the effects of HP on the production of ROS by human neutrophils between two groups-22 patients with gastric cancer and 16 patients without gastric cancer (control group), in whom HP was isolated from stomach biopsy tissues-using a luminol- and lucigenin-dependent chemiluminescence (LmCL and LgCL, respectively). It was very similar in the mean value or variance of mean maximal chemiluminescence intensities (MCI) and peak time in LmCL and LgCL between the two groups. MCI of LmCL was highly correlated with that of LgCL in both groups. These results indicate that there were no differences in the behaviour of HP on human neutrophil chemiluminescence between two groups. The progression from non-malignant mucosa to cancer may be associated with the time-dependent effects of HP via ROS produced by neutrophils on the gastric mucosa. PMID- 11038485 TI - Chemiluminometric determination of tissue polypeptide antigen (TPA), cancer antigen 15-3 (CA 15-3), carcinoembryonic antigen (CEA) in comparison with vascular endothelial growth factor (VEGF) in follow-up of breast cancer. AB - Vascular endothelial growth factor (VEGF), tissue polypeptide antigen (TPA), cancer antigen 15-3 (CA 15-3) and carcinoembryonic antigen (CEA) were measured in 314 sera of breast cancer patients and in 58 sera of women without breast cancer. VEGF was determined using a sandwich enzyme immunoassay technique (ELISA) and the tumour markers TPA, CA 15-3 and CEA with an immunoluminometric assay (ILMA). The breast cancer patients were staged according to the TNM classification stages 0 IV (by UICC) in patient groups with a compatible prognosis. Median and range of each stage were investigated. The cut-off values (95th and 97.5th percentile of control group) of VEGF, TPA, CA15-3 and CEA were determined; sensitivities for each parameter and for all combinations of two parameters were investigated for these cut-offs and the receiver operating characteristic (ROC) curves were calculated. The differences between the control group and stages 0-3 were shown to be non-significant for CA 15-3 and CEA but significant for VEGF and TPA. Significant differences were found in stage 4 for VEGF and all three markers. The increase in sensitivity of VEGF from stage 0 to stage 3 and the decrease from stage 3 to stage 4 can be interpreted based on the role of VEGF in the angiogenesis. The quantification of VEGF could give additional information for selecting patients for systemic adjuvant therapy. PMID- 11038486 TI - Detection of tetracyclines with luminescent bacterial strains. AB - The performance of two bioluminescent Escherichia coli K-12 strains for the specific detection of the tetracycline family of antimicrobial agents was compared, and the analytical applicability of one of the strains was preliminarily evaluated. One sensor plasmid contained the bacterial luciferase operon of Photorhabdus luminescens under the control of the tetracycline responsive element from transposon Tn10 (15). An analogous plasmid construction with firefly (Photinus pyralis) luciferase reporter gene was constructed, and these two divergent tetracycline-inducible light-emitting systems were compared for their suitability for the qualitative detection of tetracyclines. Both sensor strains behaved in a similar manner kinetically, and the most sensitive tetracycline response for both sensor strains was achieved in 90-120 min by performing the assay at 37 degrees C. The sensor strain containing the bacterial luciferase operon responded slightly more sensitively to different tetracyclines than the strain containing firefly luciferase gene. The sensor bacteria retained their inducibility in lyophilization, and freeze-dried cells detected tetracyclines as sensitively as freshly cultivated sensor cells. The preliminary results from the analysis of tetracycline-spiked pork serum samples indicated that these sensor bacteria could be used to screen veterinary samples for tetracycline residues in real-time. PMID- 11038487 TI - Long-lasting chemiluminescence of luminol on electrochemically pre-oxidized platinum electrodes in NaOH solution. AB - A long-lasting bright chemiluminescence (CL) of luminol was generated at polycrystalline platinum electrodes with open circuit. The CL can last for several hours with the presence of O(2) in the solution when the electrode was preoxidized at potentials more positive than 1.10 V vs. SCE. The effects of the varieties of solution conditions and surface states of the electrode on the CL intensity and the interfacial potential of the electrode were investigated. It was proposed that PtO was generated at the pre-oxidized potentials and played a role of catalyst of luminol oxidation for generating the CL. The redox couple of PtO/Pt(active) at the electrode surface was maintained in the presence of O(2) and luminol, and generated the interfacial potential more positive than 140 mV. Mathematical treatment of the reaction mechanism was conducted, which led to an approximated expression of a steady CL intensity (I(CL)) as a function of the pre polarization potential (E( h)) and time (tau( h)) of the electrode. An empirical equation, (I(CL))(4/3) = 3480(-1 + 0.82E( h) + 0.037 ln tau( h)), was estimated from the experimental data. PMID- 11038488 TI - Flow injection system with chemiluminometric detection for enzymatic determination of ascorbic acid. AB - A simple, selective and rapid method for determination of ascorbic acid from fruit juices was developed by combining a flow injection analysis (FIA) system with a chemiluminometric detector and a reactor with L-ascorbate oxidase immobilized on controlled pore glass. It was found that some reducing agents (eg ascorbic acid and mercaptoacetic acid) give chemiluminescence with luminol in the presence of hexacyanoferrate (III) in an alkaline solution. We used this new type of chemiluminescent reaction for the enzymatic determination of ascorbic acid. The background substraction method was used in order to avoid interference during ascorbic acid determination. Accordingly, two chemiluminometric signals were registered for each determination, one signal corresponding to the sample that passed through the enzymatic reactor that decomposed the ascorbic acid completely, and the second signal corresponding to the sample that does not pass through the reactor. The difference between the two signals corresponds to ascorbic acid from the sample. The linear range of the method was 10-1000 micromol/L of ascorbic acid and the detection limit was 5 micromol/L The throughput was four samples/h and RSD 3.13% (n = 10). This method was applied for determination of ascorbic in fruit juices. The results were compared with those found by the reference method, based on titrimetric determination with 2,6 dichlorophenolindophenol, and the concordance was excellent. PMID- 11038489 TI - High stability and high efficiency chemiluminescent acridinium compounds obtained from 9-acridine carboxylic esters of hydroxamic and sulphohydroxamic acids. AB - A series of hydroxamic acids and sulphohydroxamic acids were prepared and linked to 9-acridinecarboxylic acid through a pseudo-ester function. After N-methylation of the heterocyclic ring, the different compounds were tested for their chemiluminescent properties. Substituents on the hydroxamic functions have shown various effects (steric or electronic) on the luminescence yield or stability of the molecule. The most interesting derivatives were selected in terms of chemical stability and chemiluminescence efficiency. 9-[(N-hydroxysuccinimidyl-4-oxo-4-N phenylaminobutanoate)N-carb oxylat e]-10-methyl-acridinium (FA6), 9-(N phenylpivalamide-N-carboxylate)-10-methylacridinium (FA17) and 9-(N phenylpivalamide N-carboxylate)-10-carboxymethyl-acridinium (FA18) iodomercurates are very promising as chemiluminescent labels. These compounds can be detected at very low levels (10(-16)-10(-17) mol/L) and in our stability evaluation, FA6, FA17 and FA18 showed similar results to the acridinium ester DMAE. Their half lives at 20 degrees C are greater than 2 weeks. PMID- 11038490 TI - Effects of zinc on the reactive oxygen species generating capacity of human neutrophils and on the serum opsonic activity in vitro. AB - To investigate the effects of zinc on non-specific immune functions, we used the chemiluminescence method to examine the capacity of human neutrophils to produce reactive oxygen species and accompanying serum opsonic activity. When neutrophils were stimulated with both opsonized zymosan and phorbol myristate acetate in the presence of 1-10(-3) mmol/L zinc lucigenin-dependent CL responses were stable or declined, whereas luminol-dependent CL responses were significantly enhanced. The results suggest that zinc activates protein kinase C and promotes MPO degranulation and ROS metabolism, especially in hypochlorous acid production, which have the direct action of causing microbial death. Further, the lucigen dependent CL response stimulated with OZ was strongly enhanced by anti-MPO antibodies, whereas the enhancement was less in the presence of zinc, suggesting that zinc may suppress the receptor-mediated signal transduction process. Both responses were inhibited at 10 mmol/L. Serum opsonic activity was enhanced by zinc at 10(-4) and 10(-3) mmol/L but reduced at 10 mmol/L. These data indicate that addition of zinc around and above normal physiological concentrations facilitates neutrophil functional activity and serum opsonic activity, whereas these are inhibited by a lack of zinc or an excessive amount, suggesting that zinc is essential for optimal functioning of non-specific immunity. PMID- 11038521 TI - Prime factors. AB - We use Voiculescu's free probability theory to prove the existence of prime factors, hence answering a longstanding problem in the theory of von Neumann algebras. PMID- 11038522 TI - Metabolic pathway engineering in cotton: biosynthesis of polyhydroxybutyrate in fiber cells. AB - Alcaligenes eutrophus genes encoding the enzymes, beta-ketothiolase (phaA), acetoacetyl-CoA reductase (phaB), and polyhydroxyalkanoate synthase (phaC) catalyze the production of aliphatic polyester poly-d-(-)-3-hydroxybutyrate (PHB) from acetyl-CoA. PHB is a thermoplastic polymer that may modify fiber properties when synthesized in cotton. Endogenous beta-ketothiolase activity is present in cotton fibers. Hence cotton was transformed with engineered phaB and phaC genes by particle bombardment, and transgenic plants were selected based on marker gene, beta-glucuronidase (GUS), expression. Fibers of 10 transgenic plants expressed phaB gene, while eight plants expressed both phaB and phaC genes. Electron microscopy examination of fibers expressing both genes indicated the presence of electron-lucent granules in the cytoplasm. High pressure liquid chromatography, gas chromatography, and mass spectrometry evidence suggested that the new polymer produced in transgenic fibers is PHB. Sixty-six percent of the PHB in fibers is in the molecular mass range of 0.6 x 10(6) to 1.8 x 10(6) Da. The presence of PHB granules in transgenic fibers resulted in measurable changes of thermal properties. The fibers exhibited better insulating characteristics. The rate of heat uptake and cooling was slower in transgenic fibers, resulting in higher heat capacity. These data show that metabolic pathway engineering in cotton may enhance fiber properties by incorporating new traits from other genetic sources. This is an important step toward producing new generation fibers for the textile industry. PMID- 11038523 TI - Binocular vision in insects: How mantids solve the correspondence problem. AB - Praying mantids use binocular cues to judge whether their prey is in striking distance. When there are several moving targets within their binocular visual field, mantids need to solve the correspondence problem. They must select between the possible pairings of retinal images in the two eyes so that they can strike at a single real target. In this study, mantids were presented with two targets in various configurations, and the resulting fixating saccades that precede the strike were analyzed. The distributions of saccades show that mantids consistently prefer one out of several possible matches. Selection is in part guided by the position and the spatiotemporal features of the target image in each eye. Selection also depends upon the binocular disparity of the images, suggesting that insects can perform local binocular computations. The pairing rules ensure that mantids tend to aim at real targets and not at "ghost" targets arising from false matches. PMID- 11038524 TI - Convergent pathways for lipochitooligosaccharide and auxin signaling in tobacco cells. AB - Lipochitooligosaccharides (LCOs) are a novel class of plant growth regulators that activate in tobacco protoplasts the expression of AXI1, a gene implicated in auxin signaling. Transient assays with a chimeric P(AXI)-GUS expression plasmid revealed that the N-octadecenoylated monosaccharide GlcN has all structural requirements for a biological active glycolipid, whereas the inactive N-acylated GalN epimer inhibits LCO action. Specific inhibition of LCO and auxin action shows that both signals are transduced within the tobacco cell via separate pathways that converge at or before AXI1 transcription. Cytokinin is suggested to be a common effector of LCO and auxin signaling. We also show that activation of AXI1 correlates with growth factor-induced cell division. PMID- 11038525 TI - The Pto kinase mediates a signaling pathway leading to the oxidative burst in tomato. AB - The Pto gene encodes a serine/threonine kinase that confers resistance in tomato to Pseudomonas syringae pv. tomato strains that express the avirulence gene avrPto. Partial characterization of the Pto signal transduction pathway and the availability of transgenic tomato lines (+/- Pto) make this an ideal system for exploring the molecular basis of disease resistance. In this paper, we test two transgenic tomato cell suspension cultures (+/-Pto) for production of H2O2 following independent challenge with two strains of P. syringae pv. tomato (+/ avrPto). Only when Pto and avrPto are present in the corresponding organisms are two distinct phases of the oxidative burst seen, a rapid first burst followed by a slower and more prolonged second burst. In the remaining three plant-pathogen interactions, we observe either no burst or only a first burst, indicating that the second burst is correlated with disease resistance. Further support for this observation comes from the finding that both resistant and susceptible tomato lines produce the critical second oxidative burst when challenged with P. syringae pv. tabaci, a nonhost pathogen that elicits a hypersensitive response on both tomato lines. The Pto kinase is not required, however, for the oxidative burst initiated by non-specific elicitors such as oligogalacturonides or osmotic stress. A model describing a possible role for the Pto kinase in the overall scheme of oxidative burst signaling is proposed. PMID- 11038527 TI - Time-resolved optical diffusion tomographic image reconstruction in highly scattering turbid media. AB - The image of an object hidden in highly scattering media was reconstructed using a fast, noise-resistant algorithm newly applied to diffusion tomography. A pulsed light source producing scattered and transmitted light is examined at multiple times. Multiple source detector pairs around the medium are used to obtain data in many different directions. An inverse scattering algorithm with nonuniform regularization achieves rapid inversion convergence. PMID- 11038526 TI - Constitutive expression of the cold-regulated Arabidopsis thaliana COR15a gene affects both chloroplast and protoplast freezing tolerance. AB - Cold acclimation in plants is associated with the expression of COR (cold regulated) genes that encode polypeptides of unknown function. It has been widely speculated that products of these genes might have roles in freezing tolerance. Here we provide direct evidence in support of this hypothesis. We show that constitutive expression of COR15a, a cold-regulated gene of Arabidopsis thaliana that encodes a chloroplast-targeted polypeptide, enhances the in vivo freezing tolerance of chloroplasts in nonacclimated plants by almost 2 degrees C, nearly one-third of the increase that occurs upon cold acclimation of wild-type plants. Significantly, constitutive expression of COR15a also affects the in vitro freezing tolerance of protoplasts. At temperatures between -5 and -8 degrees C, the survival of protoplasts isolated from leaves of nonacclimated transgenic plants expressing COR15a was greater than that of protoplasts isolated from leaves of nonacclimated wild-type plants. At temperatures between -2 and -4 degrees C, constitutive expression of COR15a had a slight negative effect on survival. The implications of these data regarding possible modes of COR15a action are discussed. PMID- 11038528 TI - Effects of productivity, consumers, competitors, and El Nino events on food chain patterns in a rocky intertidal community. AB - We experimentally manipulated nutrient input to a rocky intertidal community, using nutrient-diffusing flowerpots, to determine (i) whether nutrients limited intertidal productivity, (ii) how a large-scale oceanographic disturbance (an El Nino event) affected patterns of nutrient limitation, (iii) the relative impacts of molluscan grazers and nutrient limitation, and (iv) if responses to experimental nutrient addition among trophic levels were more consistent with prey-dependent or ratio-dependent food chain models. Nutrients measurably increased the abundance of micrograzers (amphipods and chironomid larvae), but not algal biomass, during the summer of an El Nino year. Nutrients had no effects in two non-El Nino years and during the autumn of an El Nino year. Adding nutrients did not affect food chain stability as assessed by temporal variation in algal biomass and micrograzer abundance. Large molluscan grazers caused large reductions in micrograzers and smaller reductions in algae, indicating consistent consumer effects. The results demonstrate that in this intertidal community, nutrient limitation can occur under conditions of nutrient stress, that top-down grazing effects are typically stronger than bottom-up nutrient effects, and that prey-dependent models are more appropriate than ratio-dependent models. PMID- 11038530 TI - Arabidopsis thaliana defense-related protein ELI3 is an aromatic alcohol:NADP+ oxidoreductase. AB - We expressed a cDNA encoding the Arabidopsis thaliana defense-related protein ELI3-2 in Escherichia coli to determine its biochemical function. Based on a protein database search, this protein was recently predicted to be a mannitol dehydrogenase [Williamson, J. D., Stoop, J. M. H., Massel, M. O., Conkling, M. A. & Pharr, D. M. (1995) Proc. Natl. Acad. Sci. USA 92, 7148-7152]. Studies on the substrate specificity now revealed that ELI3-2 is an aromatic alcohol: NADP+ oxidoreductase (benzyl alcohol dehydrogenase). The enzyme showed a strong preference for various aromatic aldehydes as opposed to the corresponding alcohols. Highest substrate affinities were observed for 2-methoxybenzaldehyde, 3 methoxybenzaldehyde, salicylaldehyde, and benzaldehyde, in this order, whereas mannitol dehydrogenase activity could not be detected. These and previous results support the notion that ELI3-2 has an important role in resistance-related aromatic acid-derived metabolism. PMID- 11038532 TI - New infinite families of exact sums of squares formulas, Jacobi elliptic functions, and Ramanujan's tau function. AB - In this paper, we give two infinite families of explicit exact formulas that generalize Jacobi's (1829) 4 and 8 squares identities to 4n(2) or 4n(n + 1) squares, respectively, without using cusp forms. Our 24 squares identity leads to a different formula for Ramanujan's tau function tau(n), when n is odd. These results arise in the setting of Jacobi elliptic functions, Jacobi continued fractions, Hankel or Turanian determinants, Fourier series, Lambert series, inclusion/exclusion, Laplace expansion formula for determinants, and Schur functions. We have also obtained many additional infinite families of identities in this same setting that are analogous to the eta-function identities in appendix I of Macdonald's work [Macdonald, I. G. (1972) Invent. Math. 15, 91 143]. A special case of our methods yields a proof of the two conjectured [Kac, V. G. and Wakimoto, M. (1994) in Progress in Mathematics, eds. Brylinski, J.-L., Brylinski, R., Guillemin, V. & Kac, V. (Birkhauser Boston, Boston, MA), Vol. 123, pp. 415-456] identities involving representing a positive integer by sums of 4n(2) or 4n(n + 1) triangular numbers, respectively. Our 16 and 24 squares identities were originally obtained via multiple basic hypergeometric series, Gustafson's C(l) nonterminating (6)phi(5) summation theorem, and Andrews' basic hypergeometric series proof of Jacobi's 4 and 8 squares identities. We have (elsewhere) applied symmetry and Schur function techniques to this original approach to prove the existence of similar infinite families of sums of squares identities for n(2) or n(n + 1) squares, respectively. Our sums of more than 8 squares identities are not the same as the formulas of Mathews (1895), Glaisher (1907), Ramanujan (1916), Mordell (1917, 1919), Hardy (1918, 1920), Kac and Wakimoto, and many others. PMID- 11038529 TI - Distinct roles for two G protein alpha subunits in fungal virulence, morphology, and reproduction revealed by targeted gene disruption. AB - Reduced accumulation of the GTP-binding protein G(i)alpha subunit CPG-1, due either to hypovirus infection or transgenic cosuppression, correlates with virulence attenuation of the chestnut blight fungus, Cryphonectria parasitica. The role of G protein-mediated signal transduction in fungal virulence was further examined by targeted disruption of the gene cpg-1, encoding CPG-1, and a second Galpha gene, cpg-2, encoding the subunit CPG-2. Disruption of cpg-1 resulted in a set of phenotypic changes similar to, but more severe than, those associated with hypovirus infection. Changes included a marked reduction in fungal growth rate and loss of virulence, asexual sporulation, female fertility, and transcriptional induction of the gene lac-1, encoding the enzyme laccase. In contrast, cpg-2 disruption resulted in only slight reductions in growth rate and asexual sporulation and no significant reduction in virulence, female fertility, or lac-1 mRNA inducibility. These results provide definitive confirmation of previous correlative evidence that suggested a requirement of CPG-1-linked signaling for a number of fungal processes, including virulence and reproduction, while demonstrating that a second Galpha, CPG-2, is dispensable for these processes. They also significantly strengthen support for the apparent linkage between hypovirus-mediated disruption of G protein signal transduction and attenuation of fungal virulence. PMID- 11038533 TI - Stability of nonrelativistic quantum mechanical matter coupled to the (ultraviolet cutoff) radiation field. AB - We announce a proof of H-stability for the quantized radiation field, with ultraviolet cutoff, coupled to arbitrarily many non-relativistic quantized electrons and static nuclei. Our result holds for arbitrary atomic numbers and fine structure constant. We also announce bounds for the energy of many electrons and nuclei in a classical vector potential and for the eigenvalue sum of a one electron Pauli Hamiltonian with magnetic field. PMID- 11038534 TI - High-resolution mapping and isolation of a yeast artificial chromosome contig containing fw2.2: a major fruit weight quantitative trait locus in tomato. AB - A high-resolution physical and genetic map of a major fruit weight quantitative trait locus (QTL), fw2.2, has been constructed for a region of tomato chromosome 2. Using an F(2) nearly isogenic line mapping population (3472 individuals) derived from Lycopersicon esculentum (domesticated tomato) x Lycopersicon pennellii (wild tomato), fw2.2 has been placed near TG91 and TG167, which have an interval distance of 0.13 +/- 0.03 centimorgan. The physical distance between TG91 and TG167 was estimated to be /=4- to 5-fold and reduced contamination by other cellular membranes. For each purified fraction, the mean apparent diameter of membrane vesicles was determined by freeze-fracture electron microscopy, and the osmotic shrinking kinetics of the vesicles were characterized by stopped-flow light scattering. Osmotic water permeability coefficients (Pf) of 6.1 +/- 0.2 and 7.6 +/- 0.9 microm . s(-1) were deduced for PM-enriched vesicles purified by FFE and phase partitioning, respectively. The associated activation energies (Ea; 13.7 +/- 1.0 and 13.4 +/- 1.4 kcal . mol(-1), respectively) suggest that water transport in the purified PM occurs mostly by diffusion across the lipid matrix. In contrast, water transport in TP vesicles purified by FFE was characterized by (i) a 100-fold higher Pf of 690 +/- 35 microm . s(-1), (ii) a reduced Ea of 2.5 +/- 1.3 kcal . mol(-1), and (iii) a reversible inhibition by mercuric chloride, up to 83% at 1 mM. These results provide functional evidence for channel-mediated water transport in the TP, and more generally in a higher plant membrane. A high TP Pf suggests a role for the vacuole in buffering osmotic fluctuations occurring in the cytoplasm. Thus, the differential water permeabilities and water channel activities observed in the tobacco TP and PM point to an original osmoregulatory function for water channels in relation to the typical compartmentation of plant cells. PMID- 11038556 TI - Polycaps: reversibly formed polymeric capsules. AB - Described are assemblies consisting of polymeric capsules, "polycaps," formed from two calix[4]arene tetraureas covalently connected at their lower rims. In these structures self-assembly leads to reversibly formed capsule sites along a chain, reminiscent of beads on a string. Their dynamic behavior is characterized by 1H NMR spectroscopy through encapsulation of guest species, reversible polymerization, and the formation of sharply defined hybrid capsules. PMID- 11038557 TI - A global budget for fine root biomass, surface area, and nutrient contents. AB - Global biogeochemical models have improved dramatically in the last decade in their representation of the biosphere. Although leaf area data are an important input to such models and are readily available globally, global root distributions for modeling water and nutrient uptake and carbon cycling have not been available. This analysis provides global distributions for fine root biomass, length, and surface area with depth in the soil, and global estimates of nutrient pools in fine roots. Calculated root surface area is almost always greater than leaf area, more than an order of magnitude so in grasslands. The average C:N:P ratio in living fine roots is 450:11:1, and global fine root carbon is more than 5% of all carbon contained in the atmosphere. Assuming conservatively that fine roots turn over once per year, they represent 33% of global annual net primary productivity. PMID- 11038558 TI - In vitro reconstitution of the photosystem I light-harvesting complex LHCI-730: heterodimerization is required for antenna pigment organization. AB - Here we describe the in vitro reconstitution of photosystem I light-harvesting complexes with pigments and proteins (Lhca1 and Lhca4) obtained by overexpression of tomato Lhca genes in Escherichia coli. Using Lhca1 and Lhca4 individually for reconstitution results in monomeric pigment-proteins, whereas a combination thereof yields a dimeric complex. Interactions of the apoproteins is highly specific, as reconstitution of either of the two constituent proteins in combination with a light-harvesting protein of photosystem II does not result in dimerization. The reconstituted Lhca1/4, but not complexes obtained with either Lhca1 or Lhca4 alone, closely resembles the native LHCI-730 dimer from tomato leaves with regard to spectroscopic properties, pigment composition, and stoichiometry. Monomeric complexes of Lhca1 or Lhca4 possess lower pigment/protein ratios, indicating that interactions of the two subunits not only facilitates pigment reorganization but also recruitment of additional pigments. In addition to higher averages of chlorophyll a/b ratios in monomeric complexes than in LHCI-730, comparative fluorescence and CD spectra demonstrate that heterodimerization involves preferential ligation of more chlorophyll b. PMID- 11038559 TI - Structure of the zero-pressure-gradient turbulent boundary layer. AB - A processing of recent experimental data by Nagib and Hites [Nagib, H. & Hites, M. (1995) AIAA paper 95-0786, Reno, NV) shows that the flow in a zero-pressure gradient turbulent boundary layer, outside the viscous sublayer, consists of two self-similar regions, each described by a scaling law. The results concerning the Reynolds-number dependence of the coefficients of the wall-region scaling law are consistent with our previous results concerning pipe flow, if the proper definition of the boundary layer Reynolds number (or boundary layer thickness) is used. PMID- 11038560 TI - Factors controlling the competition among rotational and vibrational energy transfer channels in glyoxal. AB - The state-to-state transfer of rotational and vibrational energy has been studied for S1 glyoxal (CHOCHO) in collisions with D2, N2, CO and C2H4 using crossed molecular beams. A laser is used to pump glyoxal seeded in He to its S1 zero point level with zero angular momentum about its top axis (K' = 0). The inelastic scattering to each of at least 26 S1 glyoxal rotational and rovibrational levels is monitored by dispersed S1-S0 fluorescence. Various collision partners are chosen to investigate the relative influences of reduced mass and the collision pair interaction potential on the competition among the energy transfer channels. When the data are combined with that obtained previously from other collision partners whose masses range from 2 to 84 amu, it is seen that the channel competition is controlled primarily by the kinematics of the collisional interaction. Variations in the intermolecular potential play strictly a secondary role. PMID- 11038562 TI - Regulation of chloroplast protein import through a protochlorophyllide-responsive transit peptide. AB - NADPH:protochlorophyllide (Pchlide) oxidoreductase (POR) is the key enzyme of chlorophyll biosynthesis in angiosperms. In barley, two POR enzymes, termed PORA and PORB, exist. Both are nucleus-encoded plastid proteins that must be imported posttranslationally from the cytosol. Whereas the import of the precursor of PORA, pPORA, previously has been shown to depend on Pchlide, the import of pPORB occurred constitutively. To study this striking difference, chimeric precursor proteins were constructed in which the transit sequences of the pPORA and pPORB were exchanged and fused to either their cognate polypeptides or to a cytosolic dihydrofolate reductase (DHFR) reporter protein of mouse. As shown here, the transit peptide of the pPORA (transA) conferred the Pchlide requirement of import onto both the mature PORB and the DHFR. By contrast, the transit peptide of the pPORB directed the reporter protein into both chloroplasts that contained or lacked translocation-active Pchlide. In vitro binding studies further demonstrated that the transit peptide of the pPORA, but not of the pPORB, is able to bind Pchlide. We conclude that the import of the authentic pPORA and that of the transA-PORB and transA-DHFR fusion proteins is regulated by a direct transit peptide-Pchlide interaction, which is likely to occur in the plastid envelope, a major site of porphyrin biosynthesis. PMID- 11038561 TI - Characterization of the transposition pattern of the Ac element in Arabidopsis thaliana using endonuclease I-SceI. AB - We have investigated physical distances and directions of transposition of the maize transposable element Ac in Arabidopsis thaliana. We prepared a transferred DNA (T-DNA) construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-SceI (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. Three transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. These transgenic plants were crossed with the Arabidopsis that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with endonuclease I-SceI, sizes of segment of DNA were determined by pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results showed that 50% of all transposition events had occurred within 1,700 kb on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. PMID- 11038563 TI - enod40 induces dedifferentiation and division of root cortical cells in legumes. AB - Under nitrogen-limiting conditions Rhizobium meliloti can establish symbiosis with Medicago plants to form nitrogen-fixing root nodules. Nodule organogenesis starts with the dedifferentiation and division of root cortical cells. In these cells the early nodulin gene enod40, which encodes an unusually small peptide (12 or 13 amino acids), is induced from the beginning of this process. Herein we show that enod40 expression evokes root nodule initiation. (i) Nitrogen-deprived transgenic Medicago truncatula plants overexpressing enod40 exhibit extensive cortical cell division in their roots in the absence of Rhizobium. (ii) Bombardment of Medicago roots with an enod40-expressing DNA cassette induces dedifferentiation and division of cortical cells and the expression of another early nodulin gene, Msenod12A. Moreover, transient expression of either the enod40 region spanning the oligopeptide sequence or only the downstream region without this sequence induces these responses. Our results suggest that the cell specific growth response elicited by enod40 is involved in the initiation of root nodule organogenesis. PMID- 11038564 TI - Kinematic geometry of mass-triangles and reduction of Schrodinger's equation of three-body systems to partial differential equations solely defined on triangular parameters. AB - Schrodinger's equation of a three-body system is a linear partial differential equation (PDE) defined on the 9-dimensional configuration space, R9, naturally equipped with Jacobi's kinematic metric and with translational and rotational symmetries. The natural invariance of Schrodinger's equation with respect to the translational symmetry enables us to reduce the configuration space to that of a 6-dimensional one, while that of the rotational symmetry provides the quantum mechanical version of angular momentum conservation. However, the problem of maximizing the use of rotational invariance so as to enable us to reduce Schrodinger's equation to corresponding PDEs solely defined on triangular parameters--i.e., at the level of R6/SO(3)--has never been adequately treated. This article describes the results on the orbital geometry and the harmonic analysis of (SO(3),R6) which enable us to obtain such a reduction of Schrodinger's equation of three-body systems to PDEs solely defined on triangular parameters. PMID- 11038565 TI - Temporal variation, dormancy, and coexistence: a field test of the storage effect. AB - Theoretical models suggest that overlapping generations, in combination with a temporally fluctuating environment, may allow the persistence of competitors that otherwise would not coexist. Despite extensive theoretical development, this "storage effect" hypothesis has received little empirical attention. Herein I present the first explicit mathematical analysis of the contribution of the storage effect to the dynamics of competing natural populations. In Oneida Lake, NY, data collected over the past 30 years show a striking negative correlation between the water-column densities of two species of suspension-feeding zooplankton, Daphnia galeata mendotae and Daphnia pulicaria. I have demonstrated competition between these two species and have shown that both possess long-lived eggs that establish overlapping generations. Moreover, recruitment to this long lived stage varies annually, so that both daphnids have years in which they are favored (for recruitment) relative to their competitor. When the long-term population growth rates are calculated both with and without the effects of a variable environment, I show that D. galeata mendotae clearly cannot persist without the environmental variation and prolonged dormancy (i.e., storage effect) whereas D. pulicaria persists through consistently high per capita recruitment to the long-lived stage. PMID- 11038566 TI - Estimating the relative roles of top-down and bottom-up forces on insect herbivore populations: a classic study revisited. AB - Although most ecologists agree that both top-down and bottom-up forces (predation and resource limitation, respectively) act in concert to influence populations of herbivores, it has proven difficult to estimate the relative contributions of such forces in terrestrial systems. Using a combination of time-series analysis of population counts recorded over 16 years and experimental data, we present the first estimates of the relative roles of top-down and bottom-up forces on the population dynamics of two terrestrial insect herbivores on the English oak (Quercus robur). Data suggest that temporal variation in winter moth, Operophtera brumata, density is dominated by time-lagged effects of pupal predators. By comparison, spatial variation in O. brumata density is dominated by host-plant quality. Overall, top-down forces explain 34.2% of population variance, bottom-up forces explain 17.2% of population variance, and 48.6% remains unexplained. In contrast, populations of the green oak tortrix, Tortrix viridana, appear dominated by bottom-up forces. Resource limitation, expressed as intraspecific competition among larvae for oak leaves, explains 29.4% of population variance. Host quality effects explain an additional 5.7% of population variance. We detected no major top-down effects on T. viridana populations. An unknown factor causing a linear decline in T. viridana populations over the 16-year study period accounts for most of the remaining unexplained variance. We discuss the observed differences between the insect species and the utility of time-series analysis as a tool in assessing the relative importance of top-down and bottom-up forces on herbivore populations. PMID- 11038567 TI - Importance of epistasis as the genetic basis of heterosis in an elite rice hybrid. AB - The genetic basis of heterosis was investigated in an elite rice hybrid by using a molecular linkage map with 150 segregating loci covering the entire rice genome. Data for yield and three traits that were components of yield were collected over 2 years from replicated field trials of 250 F(2:3) families. Genotypic variations explained from about 50% to more than 80% of the total variation. Interactions between genotypes and years were small compared with the main effects. A total of 32 quantitative trait loci (QTLs) were detected for the four traits; 12 were observed in both years and the remaining 20 were detected in only one year. Overdominance was observed for most of the QTLs for yield and also for a few QTLs for the component traits. Correlations between marker heterozygosity and trait expression were low, indicating that the overall heterozygosity made little contribution to heterosis. Digenic interactions, including additive by additive, additive by dominance, and dominance by dominance, were frequent and widespread in this population. The interactions involved large numbers of marker loci, most of which individually were not detectable on single-locus basis; many interactions among loci were detected in both years. The results provide strong evidence that epistasis plays a major role as the genetic basis of heterosis. PMID- 11038568 TI - Interleaved concatenated codes: new perspectives on approaching the Shannon limit. AB - The last few years have witnessed a significant decrease in the gap between the Shannon channel capacity limit and what is practically achievable. Progress has resulted from novel extensions of previously known coding techniques involving interleaved concatenated codes. A considerable body of simulation results is now available, supported by an important but limited theoretical basis. This paper presents a computational technique which further ties simulation results to the known theory and reveals a considerable reduction in the complexity required to approach the Shannon limit. PMID- 11038569 TI - Buoyancy-driven, rapid exhumation of ultrahigh-pressure metamorphosed continental crust. AB - Preservation of ultrahigh-pressure (UHP) minerals formed at depths of 90-125 km require unusual conditions. Our subduction model involves underflow of a salient (250 +/- 150 km wide, 90-125 km long) of continental crust embedded in cold, largely oceanic crust-capped lithosphere; loss of leading portions of the high density oceanic lithosphere by slab break-off, as increasing volumes of microcontinental material enter the subduction zone; buoyancy-driven return toward midcrustal levels of a thin (2-15 km thick), low-density slice; finally, uplift, backfolding, normal faulting, and exposure of the UHP terrane. Sustained over approximately 20 million years, rapid ( approximately 5 mm/year) exhumation of the thin-aspect ratio UHP sialic sheet caught between cooler hanging-wall plate and refrigerating, downgoing lithosphere allows withdrawal of heat along both its upper and lower surfaces. The intracratonal position of most UHP complexes reflects consumption of an intervening ocean basin and introduction of a sialic promontory into the subduction zone. UHP metamorphic terranes consist chiefly of transformed, yet relatively low-density continental crust compared with displaced mantle material-otherwise such complexes could not return to shallow depths. Relatively rare metabasaltic, metagabbroic, and metacherty lithologies retain traces of phases characteristic of UHP conditions because they are massive, virtually impervious to fluids, and nearly anhydrous. In contrast, H2O-rich quartzofeldspathic, gneissose/schistose, more permeable metasedimentary and metagranitic units have backreacted thoroughly, so coesite and other UHP silicates are exceedingly rare. Because of the initial presence of biogenic carbon, and its especially sluggish transformation rate, UHP paragneisses contain the most abundantly preserved crustal diamonds. PMID- 11038570 TI - Reorganization of an arid ecosystem in response to recent climate change. AB - Natural ecosystems contain many individuals and species interacting with each other and with their abiotic environment. Such systems can be expected to exhibit complex dynamics in which small perturbations can be amplified to cause large changes. Here, we document the reorganization of an arid ecosystem that has occurred since the late 1970s. The density of woody shrubs increased 3-fold. Several previously common animal species went locally extinct, while other previously rare species increased. While these changes are symptomatic of desertification, they were not caused by livestock grazing or drought, the principal causes of historical desertification. The changes apparently were caused by a shift in regional climate: since 1977 winter precipitation throughout the region was substantially higher than average for this century. These changes illustrate the kinds of large, unexpected responses of complex natural ecosystems that can occur in response to both natural perturbations and human activities. PMID- 11038572 TI - Chloroplast DNA footprints of postglacial recolonization by oaks. AB - Recolonization of Europe by forest tree species after the last glaciation is well documented in the fossil pollen record. This spread may have been achieved at low densities by rare events of long-distance dispersal, rather than by a compact wave of advance, generating a patchy genetic structure through founder effects. In long-lived oak species, this structure could still be discernible by using maternally transmitted genetic markers. To test this hypothesis, a fine-scale study of chloroplast DNA (cpDNA) variability of two sympatric oak species was carried out in western France. The distributions of six cpDNA length variants were analyzed at 188 localities over a 200 x 300 km area. A cpDNA map was obtained by applying geostatistics methods to the complete data set. Patches of several hundred square kilometers exist which are virtually fixed for a single haplotype for both oak species. This local systematic interspecific sharing of the maternal genome strongly suggests that long-distance seed dispersal events followed by interspecific exchanges were involved at the time of colonization, about 10,000 years ago. PMID- 11038571 TI - Wheat cytosolic acetyl-CoA carboxylase complements an ACC1 null mutation in yeast. AB - Spores harboring an ACC1 deletion derived from a diploid Saccharomyces cerevisiae strain, in which one copy of the entire ACC1 gene is replaced with a LEU2 cassette, fail to grow. A chimeric gene consisting of the yeast GAL10 promoter, yeast ACC1 leader, wheat cytosolic acetyl-CoA carboxylase (ACCase) cDNA, and yeast ACC1 3' tail was used to complement a yeast ACC1 mutation. The complementation demonstrates that active wheat ACCase can be produced in yeast. At low concentrations of galactose, the activity of the "wheat gene" driven by the GAL10 promoter is low and ACCase becomes limiting for growth, a condition expected to enhance transgenic yeast sensitivity to wheat ACCase-specific inhibitors. An aryloxyphenoxypropionate and two cyclohexanediones do not inhibit growth of haploid yeast strains containing the yeast ACC1 gene, but one cyclohexanedione inhibits growth of the gene-replacement strains at concentrations below 0.2 mM. In vitro, the activity of wheat cytosolic ACCase produced by the gene-replacement yeast strain is inhibited by haloxyfop and cethoxydim at concentrations above 0.02 mM. The activity of yeast ACCase is less affected. The wheat plastid ACCase in wheat germ extract is inhibited by all three herbicides at concentrations below 0.02 mM. Yeast gene-replacement strains will provide a convenient system for the study of plant ACCases. PMID- 11038573 TI - Analysis of linear trade models and relation to scale economies. AB - We discuss linear Ricardo models with a range of parameters. We show that the exact boundary of the region of equilibria of these models is obtained by solving a simple integer programming problem. We show that there is also an exact correspondence between many of the equilibria resulting from families of linear models and the multiple equilibria of economies of scale models. PMID- 11038574 TI - The problem of the spreading of a liquid film along a solid surface: a new mathematical formulation. AB - A new mathematical model is proposed for the spreading of a liquid film on a solid surface. The model is based on the standard lubrication approximation for gently sloping films (with the no-slip condition for the fluid at the solid surface) in the major part of the film where it is not too thin. In the remaining and relatively small regions near the contact lines it is assumed that the so called autonomy principle holds-i.e., given the material components, the external conditions, and the velocity of the contact lines along the surface, the behavior of the fluid is identical for all films. The resulting mathematical model is formulated as a free boundary problem for the classical fourth-order equation for the film thickness. A class of self-similar solutions to this free boundary problem is considered. PMID- 11038575 TI - Paul-Straubel-Kingdon trap for true zero-point confinement of an individual ion and reservoir. AB - A modification of the Paul-Straubel trap previously described by us may profitably be operated in a Paul-Straubel-Kingdon (PSK) mode during the initial loading of an individual ion into the trap. Thereby the coating of the trap ring electrode by the atomic beam directed upon it in earlier experiments is eliminated, as is the ionization of an already trapped ion. Coating created serious problems as it spot-wise changed the work function of the ring electrode, which caused large, uncontrolled dc fields in the trap center that prevented zero point confinement. Operating the Paul-Straubel trap with a small negative bias on the ring electrode wire is all that is required to realize the PSK mode. In this mode the tiny ring trap in the center of the long, straight wire section is surrounded by a second trapping well shaped like a long, thin-walled cylindrical shell and extending to the end-caps. There, ions may be conveniently created in this well without danger of coating the ring with barium. In addition, the long second well is useful as a multi-ion reservoir. PMID- 11038576 TI - Implications of a possible clustering of highest-energy cosmic rays. AB - Recently, a possible clustering of a subset of observed ultra-high energy cosmic rays above approximately 40 EeV (4 x 10(19) eV) in pairs near the supergalactic plane was reported. We show that a confirmation of this effect would provide information on the origin and nature of these events and, in case of charged primaries, imply interesting constraints on the extragalactic magnetic field. Possible implications for the most common models of ultra-high energy cosmic ray production in the literature are discussed. PMID- 11038577 TI - The cloud-ionosphere discharge: a newly observed thunderstorm phenomenon. AB - This paper deals with a luminous electric discharge that forms in the mesospheric region between thundercloud tops and the ionosphere at 90-km altitude. These cloud-ionosphere discharges (CIs), following visual reports dating back to the 19th century, were finally imaged by a low-light TV camera as part of the "SKYFLASH" program at the University of Minnesota in 1989. Many observations were made by various groups in the period 1993-1996. The characteristics of CIs are that they have a wide range of sizes from a few kilometers up to 50 km horizontally; they extend from 40 km to nearly 90 km vertically, with an intense region near 60-70 km and streamers extending down toward cloud tops; the CIs are partly or entirely composed of vertical luminous filaments of kilometer size. The predominate color is red. The TV images show that the CIs usually have a duration less than one TV field (16.7 ms), but higher-speed photometric measurements show that they last about 3 ms, and are delayed 3 ms after an initiating cloud-ground lightning stroke; 95% of these initiating strokes are found to be "positive" i.e., carry positive charges from clouds to ground. The preference for positive initiating strokes is not understood. Theories of the formation of CIs are briefly reviewed. PMID- 11038578 TI - Ordinary representations and modular forms. PMID- 11038579 TI - Broken symmetry and critical transport properties of random metals. AB - Recent experimental data on the conductivity final sigma+(T), T --> 0, on the metallic side of the metal-insulator transition in ideally random (neutron transmutation-doped) 70Ge:Ga have shown that final sigma+(0) ~ (N - Nc)mu with mu = (1/2), confirming earlier ultra-low-temperature results for Si:P. This value is inconsistent with theoretical predictions based on diffusive classical scaling models, but it can be understood by a quantum-directed percolative filamentary amplitude model in which electronic basis states exist which have a well-defined momentum parallel but not normal to the applied electric field. The model, which is based on a new kind of broken symmetry, also explains the anomalous sign reversal of the derivative of the temperature dependence in the critical regime. PMID- 11038580 TI - Critical transport properties of random metals in large magnetic fields. AB - The threshold behavior of the transport properties of a random metal in the critical region near a metal-insulator transition is strongly affected by the measuring electromagnetic fields. In spite of the randomness, the electrical conductivity exhibits striking phase-coherent effects due to broken symmetry, which greatly sharpen the transition compared with the predictions of effective medium theories, as previously explained for electrical conductivities. Here broken symmetry explains the sign reversal of the T --> 0 magnetoconductance of the metal-insulator transition in Si(B,P), also previously not understood by effective medium theories. Finally, the symmetry-breaking features of quantum percolation theory explain the unexpectedly very small electrical conductivity temperature exponent alpha = 0.22(2) recently observed in Ni(S,Se)2 alloys at the antiferromagnetic metal-insulator transition below T = 0.8 K. PMID- 11038581 TI - Experimental evidence for a behavior-mediated trophic cascade in a terrestrial food chain. AB - Predators of herbivorous animals can affect plant populations by altering herbivore density, behavior, or both. To test whether the indirect effect of predators on plants arises from density or behavioral responses in a herbivore population, we experimentally examined the dynamics of terrestrial food chains comprised of old field plants, leaf-chewing grasshoppers, and spider predators in Northeast Connecticut. To separate the effects of predators on herbivore density from the effects on herbivore behavior, we created two classes of spiders: (i) risk spiders that had their feeding mouth parts glued to render them incapable of killing prey and (ii) predator spiders that remained unmanipulated. We found that the effect of predators on plants resulted from predator-induced changes in herbivore behavior (shifts in activity time and diet selection) rather than from predator-induced changes in grasshopper density. Neither predator nor risk spiders had a significant effect on grasshopper density relative to a control. This demonstrates that the behavioral response of prey to predators can have a strong impact on the dynamics of terrestrial food chains. The results make a compelling case to examine behavioral as well as density effects in theoretical and empirical research on food chain dynamics. PMID- 11038582 TI - The exaptive excellence of spandrels as a term and prototype. AB - In 1979, Lewontin and I borrowed the architectural term "spandrel" (using the pendentives of San Marco in Venice as an example) to designate the class of forms and spaces that arise as necessary byproducts of another decision in design, and not as adaptations for direct utility in themselves. This proposal has generated a large literature featuring two critiques: (i) the terminological claim that the spandrels of San Marco are not true spandrels at all and (ii) the conceptual claim that they are adaptations and not byproducts. The features of the San Marco pendentives that we explicitly defined as spandrel-properties-their necessary number (four) and shape (roughly triangular)-are inevitable architectural byproducts, whatever the structural attributes of the pendentives themselves. The term spandrel may be extended from its particular architectural use for two dimensional byproducts to the generality of "spaces left over," a definition that properly includes the San Marco pendentives. Evolutionary biology needs such an explicit term for features arising as byproducts, rather than adaptations, whatever their subsequent exaptive utility. The concept of biological spandrels including the examples here given of masculinized genitalia in female hyenas, exaptive use of an umbilicus as a brooding chamber by snails, the shoulder hump of the giant Irish deer, and several key features of human mentality-anchors the critique of overreliance upon adaptive scenarios in evolutionary explanation. Causes of historical origin must always be separated from current utilities; their conflation has seriously hampered the evolutionary analysis of form in the history of life. PMID- 11038583 TI - Comparison of structures and energies of CH5(2+*) with CH4(+*) and their possible role in superacidic methane activation. AB - Contrary to previous theoretical studies at the UHF/6-31G* level, the methonium radical dication CH5(2+) is not a Cs symmetrical structure with a 2e-3c bond but a C2v symmetrical structure 1 with two 2e-3c bonds (at the UHF/6-31G**, UMP2/6 31G**, and UQCISD(T)/6-311G** levels). The Cs symmetrical structure is not even a minimum at the higher level of calculations. The four hydrogen atoms in 1 are bonded to the carbon atom by two 2e-3c bonds and the fifth hydrogen atom by a 2e 2c bond. The unpaired electron of 1 is located in a formal p-orbital (of the sp2 hybridized carbon atom) perpendicular to the plane of the molecule. Hydrogen scrambling in 1 is however extremely facile, as is in other C1 cations. It is found that the protonation of methane to CH5(+) decreases the energy for subsequent homolytic cleavage resulting in the exothermic (24.1 kcal/mol) formation of CH4(+*). Subsequent reaction with neutral methane while reforming CH5(+) gives the methyl radical enabling reaction with excess methane to ethane and H2. The overall reaction is endothermic by 11.4 kcal/mol, but offers under conditions of oxidative removal of H2 an alternative to the more energetic carbocationic conversion of methane. PMID- 11038584 TI - Efficient exchange of the primary quinone acceptor Q(A) in isolated reaction centers of Rhodopseudomonas viridis. AB - A key step in the conversion of solar energy into chemical energy by photosynthetic reaction centers (RCs) occurs at the level of the two quinones, Q(A) and Q(B), where electron transfer couples to proton transfer. A great deal of our understanding of the mechanisms of these coupled reactions relies on the seminal work of Okamura et al. [Okamura, M. Y., Isaacson, R. A., & Feher, G. (1975) Proc. Natl. Acad. Sci. USA 88, 3491-3495], who were able to extract with detergents the firmly bound ubiquinone Q(A) from the RC of Rhodobacter sphaeroides and reconstitute the site with extraneous quinones. Up to now a comparable protocol was lacking for the RC of Rhodopseudomonas viridis despite the fact that its Q(A) site, which contains 2-methyl-3-nonaprenyl-1,4 naphthoquinone (menaquinone-9), has provided the best x-ray structure available. Fourier transform infrared difference spectroscopy, together with the use of isotopically labeled quinones, can probe the interaction of Q(A) with the RC protein. We establish that a simple incubation procedure of isolated RCs of Rp. viridis with an excess of extraneous quinone allows the menaquinone-9 in the Q(A) site to be almost quantitatively replaced either by vitamin K(1), a close analogue of menaquinone-9, or by ubiquinone. To our knowledge, this is the first report of quinone exchange in bacterial photosynthesis. The Fourier transform infrared data on the quinone and semiquinone vibrations show a close similarity in the bonding interactions of vitamin K(1) with the protein at the Q(A) site of Rp. viridis and Rb. sphaeroides, whereas for ubiquinone these interactions are significantly different. The results are interpreted in terms of slightly inequivalent quinone-protein interactions by comparison with the crystallographic data available for the Q(A) site of the two RCs. PMID- 11038585 TI - Selective maintenance of allozyme differences among sympatric host races of the apple maggot fly. AB - Whether phytophagous insects can speciate in sympatry when they shift and adapt to new host plants is a controversial question. One essential requirement for sympatric speciation is that disruptive selection outweighs gene flow between insect populations using different host plants. Empirical support for host related selection (i.e., fitness trade-offs) is scant, however. Here, we test for host-dependent selection acting on apple (Malus pumila)- and hawthorn (Crataegus spp.)-infesting races of Rhagoletis pomonella (Diptera: Tephritidae). In particular, we examine whether the earlier fruiting phenology of apple trees favors pupae in deeper states of diapause (or with slower metabolisms/development rates) in the apple fly race. By experimentally lengthening the time period preceding winter, we exposed hawthorn race pupae to environmental conditions typically faced by apple flies. This exposure induced a significant genetic response at six allozyme loci in surviving hawthorn fly adults toward allele frequencies found in the apple race. The sensitivity of hawthorn fly pupae to extended periods of warm weather therefore selects against hawthorn flies that infest apples and helps to maintain the genetic integrity of the apple race by counteracting gene flow from sympatric hawthorn populations. Our findings confirm that postzygotic reproductive isolation can evolve as a pleiotropic consequence of host-associated adaptation, a central tenet of nonallopatric speciation. They also suggest that one reason for the paucity of reported fitness trade-offs is a failure to consider adequately costs associated with coordinating an insect's life cycle with the phenology of its host plant. PMID- 11038586 TI - Accelerated evolution as a consequence of transitions to mutualism. AB - Differential rates of nucleotide substitutions among taxa are a common observation in molecular phylogenetic studies, yet links between rates of DNA evolution and traits or behaviors of organisms have proved elusive. Likelihood ratio testing is used here for the first time to evaluate specific hypotheses that account for the induction of shifts in rates of DNA evolution. A molecular phylogenetic investigation of mutualist (lichen-forming fungi and fungi associated with liverworts) and nonmutualist fungi revealed four independent transitions to mutualism. We demonstrate a highly significant association between mutualism and increased rates of nucleotide substitutions in nuclear ribosomal DNA, and we demonstrate that a transition to mutualism preceded the rate acceleration of nuclear ribosomal DNA in these lineages. Our results suggest that the increased rate of evolution after the adoption of a mutualist lifestyle is generalized across the genome of these mutualist fungi. PMID- 11038587 TI - Electrophilic nitration of alkanes with nitronium hexafluorophosphate. AB - Nitration of alkanes such as methane, ethane, propane, n-butane, isobutane, neopentane, and cyclohexane was carried out with nitronium hexafluorophosphate in methylene chloride or nitroethane solution. Nitration of methane, albeit in poor yield, required protolytic activation of the nitronium ion. The results indicate direct electrophilic insertion of NO2+ into C-H and C-C final sigma-bonds. PMID- 11038588 TI - Phalangeal curvature and positional behavior in extinct sloth lemurs (Primates, Palaeopropithecidae). AB - Recent paleontological discoveries in Madagascar document the existence of a diverse clade of palaeopropithecids or "sloth lemurs": Mesopropithecus (three species), Babakotia (one species), Palaeopropithecus (three species), and Archaeoindris (one species). This mini-radiation of now extinct ("subfossil") lemurs is most closely related to the living indrids (Indri, Propithecus, and Avahi). Whereas the extant indrids are known for their leaping acrobatics, the palaeopropithecids (except perhaps for the poorly known giant Archaeoindris) exhibit numerous skeletal design features for antipronograde or suspensory positional behaviors (e.g., high intermembral indices and mobile joints). Here we analyze the curvature of the proximal phalanges of the hands and feet. Computed as the included angle (theta), phalangeal curvature develops in response to mechanical use and is known to be correlated in primates with hand and foot function in different habitats; terrestrial species have straighter phalanges than their arboreal counterparts, and highly suspensory forms such as the orangutan possess the most curved phalanges. Sloth lemurs as a group are characterized by very curved proximal phalanges, exceeding those seen in spider monkeys and siamangs, and approaching that of orangutans. Indrids have curvatures roughly half that of sloth lemurs, and the more terrestrial, subfossil Archaeolemur possesses the least curved phalanges of all the indroids. Taken together with many other derived aspects of their postcranial anatomy, phalangeal curvature indicates that the sloth lemurs are one of the most suspensory clades of mammals ever to evolve. PMID- 11038589 TI - Endosperm balance number manipulation for direct in vivo germplasm introgression to potato from a sexually isolated relative (Solanum commersonii Dun.). AB - Diploid (2n = 2x = 24) Solanum species with endosperm balance number (EBN) = 1 are sexually isolated from diploid 2EBN species and both tetraploid (2n = 4x = 48, 4EBN) and haploid (2n = 2x = 24, 2EBN) S. tuberosum Group Tuberosum. To sexually overcome these crossing barriers in the diploid species S. commersonii (1EBN), the manipulation of the EBN was accomplished by scaling up and down ploidy levels. Triploid F1 hybrids between an in vitro-doubled clone of S. commersonii (2n = 4x = 48, 2EBN) and diploid 2EBN clones were successfully used in 3x x 4x crosses with S. tuberosum Group Tuberosum, resulting in pentaploid/near pentaploid BC1 progenies. This provided evidence of 2n (3x) egg formation in the triploid female parents. Two selected BC1 pentaploid hybrids were successfully backcrossed both as male and as female parents with S. tuberosum Group Tuberosum. The somatic chromosome number varied greatly among the resulting BC2 progenies, which included hyperaneuploids, but also a number (4.8%) of 48-chromosome plants. The introgression of S. commersonii genomes was confirmed by the presence of S. commersonii-specific randomly amplified polymorphic DNA markers in the BC2 population analyzed. The results clearly demonstrate the feasibility of germplasm introgression from sexually isolated diploid 1EBN species into the 4x (4EBN) gene pool of the cultivated potato using sexual hybridization. Based on the amount and type of genetic variation generated, cumbersomeness, general applicability, costs, and other factors, it would be interesting to compare the approach reported here with other in vitro or in vivo, direct or indirect, approaches previously reported. PMID- 11038590 TI - Third-generation synchrotron x-ray diffraction of 6-microm crystal of raite, approximately Na3Mn3Ti0.25Si8O20(OH)2.10H2O, opens up new chemistry and physics of low-temperature minerals. AB - The crystal structure of raite was solved and refined from data collected at Beamline Insertion Device 13 at the European Synchrotron Radiation Facility, using a 3 x 3 x 65 microm single crystal. The refined lattice constants of the monoclinic unit cell are a = 15.1(1) A; b = 17.6(1) A; c = 5.290(4) A; beta = 100.5(2) degrees; space group C2/m. The structure, including all reflections, refined to a final R = 0.07. Raite occurs in hyperalkaline rocks from the Kola peninsula, Russia. The structure consists of alternating layers of a hexagonal chicken-wire pattern of 6-membered SiO4 rings. Tetrahedral apices of a chain of Si six-rings, parallel to the c-axis, alternate in pointing up and down. Two six ring Si layers are connected by edge-sharing octahedral bands of Na+ and Mn3+ also parallel to c. The band consists of the alternation of finite Mn-Mn and Na Mn-Na chains. As a consequence of the misfit between octahedral and tetrahedral elements, regions of the Si-O layers are arched and form one-dimensional channels bounded by 12 Si tetrahedra and 2 Na octahedra. The channels along the short c axis in raite are filled by isolated Na(OH,H2O)6 octahedra. The distorted octahedrally coordinated Ti4+ also resides in the channel and provides the weak linkage of these isolated Na octahedra and the mixed octahedral tetrahedral framework. Raite is structurally related to intersilite, palygorskite, sepiolite, and amphibole. PMID- 11038591 TI - Isolation and characterization of neutral-lipid-containing organelles and globuli filled plastids from Brassica napus tapetum. AB - The monolayer tapetum cells of the maturing flowers of Brassica napus contain abundant subcellular globuli-filled plastids and special lipid particles, both enriched with lipids that are supposed to be discharged and deposited onto the surface of adjacent maturing pollen. We separated the two organelles by flotation density gradient centrifugation and identified them by electron microscopy. The globuli-filled plastids had a morphology similar to those described in other plant species and tissues. They had an equilibrium density of 1.02 g/cm(3) and contained neutral esters and unique polypeptides. The lipid particles contained patches of osmiophilic materials situated among densely packed vesicles and did not have an enclosing membrane. They exhibited osmotic properties, presumably exerted by the individual vesicles. They had an equilibrium density of 1.05 g/cm(3) and possessed triacylglycerols and unique polypeptides. Several of these polypeptides were identified, by their N-terminal sequences or antibody cross reactivity, as oleosins, proteins known to be associated with seed storage oil bodies. The morphological and biochemical characteristics of the lipid particles indicate that they are novel organelles in eukaryotes that have not been previously isolated and studied. After lysis of the tapetum cells at a late stage of floral development, only the major plastid neutral ester was recovered, whereas the other abundant lipids and proteins of the two tapetum organelles were present in fragmented forms or absent on the pollen surface. PMID- 11038592 TI - Inhibition of phospholipase D by lysophosphatidylethanolamine, a lipid-derived senescence retardant. AB - Phospholipid signaling mediated by lipid-derived second messengers or biologically active lipids is still new and is not well established in plants. We recently have found that lysophosphatidylethanolamine (LPE), a naturally occurring lipid, retards senescence of leaves, flowers, and postharvest fruits. Phospholipase D (PLD) has been suggested as a key enzyme in mediating the degradation of membrane phospholipids during the early stages of plant senescence. Here we report that LPE inhibited the activity of partially purified cabbage PLD in a cell-free system in a highly specific manner. Inhibition of PLD by LPE was dose-dependent and increased with the length and unsaturation of the LPE acyl chain whereas individual molecular components of LPE such as ethanolamine and free fatty acid had no effect on PLD activity. Enzyme-kinetic analysis suggested noncompetitive inhibition of PLD by LPE. In comparison, the related lysophospholipids such as lysophosphatidylcholine, lysophosphatidylglycerol, and lysophosphotidylserine had no significant effect on PLD activity whereas PLD was stimulated by lysophosphatidic acid and inhibited by lysophosphatidylinositol. Membrane-associated and soluble PLD, extracted from cabbage and castor bean leaf tissues, also was inhibited by LPE. Consistent with acyl-specific inhibition of PLD by LPE, senescence of cranberry fruits as measured by ethylene production was more effectively inhibited according to the increasing acyl chain length and unsaturation of LPE. There are no known specific inhibitors of PLD in plants and animals. We demonstrate specific inhibitory regulation of PLD by a lysophospholipid. PMID- 11038593 TI - The influence of the flow of the reacting gas on the conditions for a thermal explosion. AB - The classical problem of thermal explosion is modified so that the chemically active gas is not at rest but is flowing in a long cylindrical pipe. Up to a certain section the heat-conducting walls of the pipe are held at low temperature so that the reaction rate is small and there is no heat release; at that section the ambient temperature is increased and an exothermic reaction begins. The question is whether a slow reaction regime will be established or a thermal explosion will occur. The mathematical formulation of the problem is presented. It is shown that when the pipe radius is larger than a critical value, the solution of the new problem exists only up to a certain distance along the axis. The critical radius is determined by conditions in a problem with a uniform axial temperature. The loss of existence is interpreted as a thermal explosion; the critical distance is the safe reactor's length. Both laminar and developed turbulent flow regimes are considered. In a computational experiment the loss of the existence appears as a divergence of a numerical procedure; numerical calculations reveal asymptotic scaling laws with simple powers for the critical distance. PMID- 11038594 TI - Twist grain boundaries in three-dimensional lamellar Turing structures. AB - Steady spatial self-organization of three-dimensional chemical reaction-diffusion systems is discussed with the emphasis put on the possible defects that may alter the Turing patterns. It is shown that one of the stable defects of a three dimensional lamellar Turing structure is a twist grain boundary embedding a Scherk minimal surface. PMID- 11038595 TI - On the geometry of solutions of the quasi-geostrophic and Euler equations. AB - We study solutions of the two-dimensional quasi-geostrophic thermal active scalar equation involving simple hyperbolic saddles. There is a naturally associated notion of simple hyperbolic saddle breakdown. It is proved that such breakdown cannot occur in finite time. At large time, these solutions may grow at most at a quadruple-exponential rate. Analogous results hold for the incompressible three dimensional Euler equation. PMID- 11038596 TI - Filamentary microstructure and linear temperature dependence of normal state transport in optimized high temperature superconductors. AB - A filamentary model of "metallic" conduction in layered high temperature superconductive cuprates explains the concurrence of normal state resistivities (Hall mobilities) linear in T (T-2) with optimized superconductivity. The model predicts the lowest temperature T0 for which linearity holds and it also predicts the maximum superconductive transition temperature Tc. The theory abandons the effective medium approximation that includes Fermi liquid as well as all other nonpercolative models in favor of countable smart basis states. PMID- 11038597 TI - High-precision position-specific isotope analysis. AB - Intramolecular carbon isotope distributions reflect details of the origin of organic compounds and may record the status of complex systems, such as environmental or physiological states. A strategy is reported here for high precision determination of 13C/12C ratios at specific positions in organic compounds separated from complex mixtures. Free radical fragmentation of methyl palmitate, a test compound, is induced by an open tube furnace. Two series of peaks corresponding to bond breaking from each end of the molecule are analyzed by isotope ratio mass spectrometry and yield precisions of SD(delta-13C) < 0.4 per thousand. Isotope labeling in the carboxyl, terminal, and methyl positions demonstrates the absence of rearrangement during activation and fragmentation. Negligible isotopic fractionation was observed as degree of fragmentation was adjusted by changing pyrolysis temperature. [1-13C]methyl palmitate with overall delta-13C = 4.06 per thousand, yielded values of +457 per thousand for the carboxyl position, in agreement with expectations from the dilution, and an average of -27.95 per thousand for the rest of the molecule, corresponding to 27.46 per thousand for the olefin series. These data demonstrate the feasibility of automated high-precision position-specific analysis of carbon for molecules contained in complex mixtures. PMID- 11038598 TI - Using a parity-sensitive sieve to count prime values of a polynomial. AB - It is expected that any irreducible polynomial with integer coefficients assumes infinitely many prime values provided that it satisfies some obvious local conditions. Moreover, it is expected that the frequency of these primes obeys a simple asymptotic law. This has however been proven for only a few special classes of polynomials. In the most famous unsolved cases the sequence of values is "thin" in the sense that it contains fewer than N(theta) integers up to N for some constant theta < 1. Quite generally it seems to be difficult to show the infinitude of primes in a given thin integer sequence and there is no polynomial for which this has hitherto been done. The polynomial x(2) + y(4) is an example of such a thin sequence; here, specifically, theta = 3/4. We report here the development of new methods that rigorously demonstrate the asymptotic formula in the case of this polynomial and that are applicable to an infinite class of polynomials to which this one belongs. The proof is based partly on a new sieve method that breaks the well-known parity problem of sieve theory and partly on a careful harmonic analysis of the special properties of biquadratic polynomial sequences. PMID- 11038599 TI - Single-chain fusions of two unrelated homeodomain proteins trigger pathogenicity in Ustilago maydis. AB - Pathogenic and sexual development of the fungus Ustilago maydis, the causal agent of corn smut disease, is regulated by heterodimerization of two unrelated homeodomain proteins bE and bW, both encoded by the multi-allelic b mating-type locus. This complex can only be formed if the two proteins are derived from different alleles. The heterodimer is believed to function as a transcriptional regulator that binds to target sites upstream of developmentally regulated genes. We have synthesized a translational fusion in which bE is tethered to bW by a designed flexible kink region. U. maydis strains expressing this synthetic b fusion become pathogenic for corn illustrating that the single-chain fusion substitutes for the active bE/bW heterodimer. Synthetic b-fusions in which bE and bW originate from the same allele as well as fusions deleted for the dimerization domains were shown to be active while both homeodomains were required for function. Such active fusion proteins are expected to be instrumental in the identification of pathogenicity genes. PMID- 11038600 TI - Scaling regions for food web properties. AB - The robustness of eight common food web properties is examined with respect to web size. We show that the current controversy concerning the scale dependence or scale invariance of these properties can be resolved by accounting for scaling constraints introduced by webs of very small size. We demonstrate statistically that the most robust way to view these properties is not to lump webs of all sizes, but to divide them into two distinct categories. For the present data set, small webs containing 12 or fewer species exhibit scale dependence, and larger webs containing more than 12 species exhibit scale invariance. PMID- 11038601 TI - Strong density- and diversity-related effects help to maintain tree species diversity in a neotropical forest. AB - Intraspecific density-dependent effects in the Barro Colorado Island (Panama) study area are far stronger, and involve far more species, than previously had been suspected. Significant effects on recruitment, many extremely strong, are seen for 67 out of the 84 most common species in the plot, including the 10 most common. Significant effects on the intrinsic rate of increase are seen in 54 of the 84 species. These effects are far more common than interspecific effects, and are predominantly of the type that should maintain tree diversity. As a result, the more diverse an area in the forest is, the higher is the overall rate of increase of the trees in that area, although sheer crowding has by itself a negative effect. These findings are consistent with, but do not prove, an important role for host-pathogen interactions (defined broadly) in the maintenance of diversity. Ways are suggested by which to test host-pathogen models and competing models. PMID- 11038602 TI - Mechanism of photosystem II photoinactivation and D1 protein degradation at low light: the role of back electron flow. AB - Light intensities that limit electron flow induce rapid degradation of the photosystem II (PSII) reaction center D1 protein. The mechanism of this phenomenon is not known. We propose that at low excitation rates back electron flow and charge recombination between the QB*- or QA*- semiquinone acceptors and the oxidized S(2,3) states of the PSII donor side may cause oxidative damage via generation of active oxygen species. Therefore, damage per photochemical event should increase with decreasing rates of PSII excitation. To test this hypothesis, the effect of the dark interval between single turnover flashes on the inactivation of water oxidation, charge separation and recombination, and the degradation of D1 protein were determined in spinach thylakoids. PSII inactivation per flash increases as the dark interval between the flashes increases, and a plateau is reached at dark intervals, allowing complete charge recombination of the QB*-/S2,3 or QA*-/S2 states (about 200 and 40 s, respectively). At these excitation rates: (i) 0.7% and 0.4% of PSII is inactivated and 0.4% and 0.2% of the D1 protein is degraded per flash, respectively, and (ii) the damage per flash is about 2 orders of magnitude higher than that induced by equal amount of energy delivered by excess continuous light. No PSII damage occurs if flashes are given in anaerobic conditions. These results demonstrate that charge recombination in active PSII is promoted by low rates of excitation and may account for a the high quantum efficiency of the rapid turnover of the D1 protein induced by limiting light. PMID- 11038603 TI - Plastoquinol at the quinol oxidation site of reduced cytochrome bf mediates signal transduction between light and protein phosphorylation: thylakoid protein kinase deactivation by a single-turnover flash. AB - Redox-controlled phosphorylation of thylakoid membrane proteins represents a unique system for the regulation of light energy utilization in photosynthesis. The molecular mechanisms for this process remain unknown, but current views suggest that the plastoquinone pool directly controls the activation of the kinase. On the basis of enzyme activation by a pH shift in the darkness combined with flash photolysis, EPR, and optical spectroscopy we propose that activation occurs when plastoquinol occupies the quinol-oxidation (Qo) site of the cytochrome bf complex, having its high-potential path components in a reduced state. A linear correlation between kinase activation and accessibility of the Qo site to plastoquinol was established by quantification of the shift in the g(y) EPR signal of the Rieske Fe-S center resulting from displacement of the Qo-site plastoquinol by a quinone analog. Activity persists as long as one plastoquinol per cytochrome bf is still available. Withdrawal of one electron from this plastoquinol after a single-turnover flash exciting photosystem I leads to deactivation of the kinase parallel with a decrease in the g(z) EPR signal of the reduced Rieske Fe-S center. Cytochrome f, plastocyanin, and P(700) are rereduced after the flash, indicating that the plastoquinol at the Qo site is limiting in maintaining the kinase activity. These results give direct evidence for a functional cytochrome bf-kinase interaction, analogous to a signal transduction system where the cytochrome bf is the receptor and the ligand is the plastoquinol at the Qo site. PMID- 11038604 TI - Redox chains in chloroplast envelope membranes: spectroscopic evidence for the presence of electron carriers, including iron-sulfur centers. AB - We have shown that envelope membranes from spinach chloroplasts contain (i) semiquinone and flavosemiquinone radicals, (ii) a series of iron-containing electron-transfer centers, and (iii) flavins (mostly FAD) loosely associated with proteins. In contrast, we were unable to detect any cytochrome in spinach chloroplast envelope membranes. In addition to a high spin [1Fe]3+ type protein associated with an EPR signal at g = 4.3, we observed two iron-sulfur centers, a [4Fe-4S]1+ and a [2Fe-2S]1+, associated with features, respectively, at g = 1.921 and g = 1.935, which were detected after reduction by NADPH and NADH, respectively. The [4Fe-4S] center, but not the [2Fe-2S] center, was also reduced by dithionite or 5-deazaflavin/oxalate. An unusual Fe-S center, named X, associated with a signal at g = 2.057, was also detected, which was reduced by dithionite but not by NADH or NADPH. Extremely fast spin-relaxation rates of flavin- and quinone-free radicals suggest their close proximity to the [4Fe-4S] cluster or the high-spin [1Fe]3+ center. Envelope membranes probably contain enzymatic activities involved in the formation and reduction of semiquinone radicals (quinol oxidase, NADPH-quinone, and NADPH-semiquinone reductases). The physiological significance of our results is discussed with respect to (i) the presence of desaturase activities in envelope membranes and (ii) the mechanisms involved in the export of protons to the cytosol, which partially regulate the stromal pH during photosynthesis. The characterization of such a wide variety of electron carriers in envelope membranes opens new fields of research on the functions of this membrane system within the plant cell. PMID- 11038605 TI - Analysis of highly disfavored processes through pathway-specific correlated fluorescence. AB - A new method is described for the detection of disfavored reaction pathways. The approach combines organic synthesis, to independently prepare reactant and product, with the low detection limits of confocally adjusted fluorescence correlation spectroscopy. Selective detection of disfavored products is achieved by designing a system in which the analyte displays a unique absorption and emission of light. This was accomplished through application of a substance selective intramolecular charge transfer. The power of this technique was demonstrated by monitoring the progress of the thermal retro-[2+2]-cycloaddition. In conjunction with concurrent 1H-NMR monitoring, the relative abundance of major and disfavored reaction products can be determined and used to calculate the energetics of processes disfavored by more than 5 kcal/mol. PMID- 11038606 TI - Plant diversity and ecosystem productivity: theoretical considerations. AB - Ecosystem processes are thought to depend on both the number and identity of the species present in an ecosystem, but mathematical theory predicting this has been lacking. Here we present three simple models of interspecific competitive interactions in communities containing various numbers of randomly chosen species. All three models predict that, on average, productivity increases asymptotically with the original biodiversity of a community. The two models that address plant nutrient competition also predict that ecosystem nutrient retention increases with biodiversity and that the effects of biodiversity on productivity and nutrient retention increase with interspecific differences in resource requirements. All three models show that both species identity and biodiversity simultaneously influence ecosystem functioning, but their relative importance varies greatly among the models. This theory reinforces recent experimental results and shows that effects of biodiversity on ecosystem functioning are predicted by well-known ecological processes. PMID- 11038607 TI - Selection of marker-free transgenic plants using the isopentenyl transferase gene. AB - We have developed a new plant vector system for repeated transformation (called MAT for multi-auto-transformation) in which a chimeric ipt gene, inserted into the transposable element Ac, is used as a selectable marker for transformation. Selectable marker genes conferring antibiotic or herbicide resistance, used to introduce economically valuable genes into crop plants, have three major problems: (i) the selective agents have negative effects on proliferation and differentiation of plant cells; (ii) there is uncertainty regarding the environmental impact of many selectable marker genes; (iii) it is difficult to perform recurrent transformations using the same selectable marker to pyramid desirable genes. The MAT vector system containing the ipt gene and the Ac element is designed to overcome these difficulties. When tobacco leaf segments were transformed and selected, subsequent excision of the modified Ac produced marker free transgenic tobacco plants without sexual crosses or seed production. In addition, the chimeric ipt gene could be visually used as a selectable marker for transformation of hybrid aspen (Populus sieboldii x Populus grandidentata). The chimeric ipt gene, therefore, is an attractive alternative to the most widely used selectable marker genes. The MAT vector system provides a promising way to shorten breeding time for genetically engineered crops. This method could be particularly valuable for fruit and forest trees, for which long generation times are a more significant barrier to breeding and genetic analysis. PMID- 11038608 TI - Cell proliferation and hair tip growth in the Arabidopsis root are under mechanistically different forms of redox control. AB - We provide evidence that the tripeptide thiol glutathione (GSH) participates in the regulation of cell division in the apical meristem of Arabidopsis roots. Exogenous application of micromolar concentrations of GSH raised the number of meristematic cells undergoing mitosis, while depletion of GSH had the opposite effect. A role for endogenous GSH in the control of cell proliferation is also provided by mapping of GSH levels in the root meristem using the GSH-specific dye monochlorobimane and confocal laser scanning microscopy. High levels of GSH were associated with the epidermal and cortical initials and markedly lower levels in the quiescent center. The mechanisms controlling cell division could also be triggered by other reducing agents: ascorbic acid and dithiothreitol. Our data also reveal significant plasticity in the relationship between the trichoblast cell length and the hair it subtends in response to alterations in intracellular redox homeostasis. While mechanisms that control trichoblast elongation are influenced by nonspecific redox couples, root hair tip growth has a more specific requirement for sulfhydryl groups. The responses we describe here may represent the extremes of redox control of root plasticity and would allow the root to maintain exploration of the soil under adverse conditions with minimal cell divisions and root hair production or capitalize on a favorable environment by production of numerous long hairs. Redox sensing of the environment and subsequent redox-dependent modulation of growth and development may be crucial components in the strategies plants have evolved for survival in a fluctuating environment. PMID- 11038609 TI - Receptor-mediated activation of a plant Ca2+-permeable ion channel involved in pathogen defense. AB - Pathogen recognition at the plant cell surface typically results in the initiation of a multicomponent defense response. Transient influx of Ca2+ across the plasma membrane is postulated to be part of the signaling chain leading to pathogen resistance. Patch-clamp analysis of parsley protoplasts revealed a novel Ca2+-permeable, La3+-sensitive plasma membrane ion channel of large conductance (309 pS in 240 mM CaCl2). At an extracellular Ca2+ concentration of 1 mM, which is representative of the plant cell apoplast, unitary channel conductance was determined to be 80 pS. This ion channel (LEAC, for large conductance elicitor activated ion channel) is reversibly activated upon treatment of parsley protoplasts with an oligopeptide elicitor derived from a cell wall protein of Phytophthora sojae. Structural features of the elicitor found previously to be essential for receptor binding, induction of defense-related gene expression, and phytoalexin formation are identical to those required for activation of LEAC. Thus, receptor-mediated stimulation of this channel appears to be causally involved in the signaling cascade triggering pathogen defense in parsley. PMID- 11038610 TI - Ethylene can stimulate Arabidopsis hypocotyl elongation in the light. AB - Ethylene inhibits hypocotyl elongation in etiolated Arabidopsis seedlings. However, when Arabidopsis was grown in the light in the presence of ethylene or its precursor 1-aminocyclopropane-1-carboxylic acid (ACC), a marked induction of hypocotyl elongation occurred. This resulted from an increase in cell expansion rather than cell division. The effects of ethylene and ACC were antagonized by the ethylene action inhibitor Ag+. The elongation response was absent or weakened in a set of ethylene-insensitive mutants (etr1-3, ein2-1, ein3-1, ein4, ain1-10, ein7). With the exception of ein4, the degree of inhibition of hypocotyl elongation was correlated with the strength of the ethylene-insensitive phenotype based on the triple response assay. In addition, the constitutive ethylene response mutant ctr1-1, grown in the light, had a longer hypocotyl than the wild type. Exogenous auxin also induced hypocotyl elongation in light-grown Arabidopsis. Again, the response was abolished by treatment with Ag+, suggesting that ethylene might be a mediator. The results showed that, depending on light conditions, ethylene can induce opposite effects on cell expansion in Arabidopsis hypocotyls. PMID- 11038611 TI - Chlorophyll a dimer: a possible primary electron donor for the photosystem II. AB - In the photosynthetic membrane, there is a particular aggregated state for the chlorophyll a (Chl a) molecules with a specific arrangement responsible for the high efficiency of energy conversion. Chl a monolayers, transferred onto solid substrates, are systems that potentially can mimic the packing of the in vivo system. The association of Chl a in the monolayer results in the formation of dimers with an average size of 3.00 +/- 0.15 nm. Considering the organization of the dimers, we assume that P680 is a dimer with the (anti) parallel transition moments of the constituent. The Chl a macrocycles most likely are tilted to each other by 30 degrees with respect to the membrane plane. PMID- 11038612 TI - Not all (possibly) "random" sequences are created equal. AB - The need to assess the randomness of a single sequence, especially a finite sequence, is ubiquitous, yet is unaddressed by axiomatic probability theory. Here, we assess randomness via approximate entropy (ApEn), a computable measure of sequential irregularity, applicable to single sequences of both (even very short) finite and infinite length. We indicate the novelty and facility of the multidimensional viewpoint taken by ApEn, in contrast to classical measures. Furthermore and notably, for finite length, finite state sequences, one can identify maximally irregular sequences, and then apply ApEn to quantify the extent to which given sequences differ from maximal irregularity, via a set of deficit (def(m)) functions. The utility of these def(m) functions which we show allows one to considerably refine the notions of probabilistic independence and normality, is featured in several studies, including (i) digits of e, pi, radical2, and radical3, both in base 2 and in base 10, and (ii) sequences given by fractional parts of multiples of irrationals. We prove companion analytic results, which also feature in a discussion of the role and validity of the almost sure properties from axiomatic probability theory insofar as they apply to specified sequences and sets of sequences (in the physical world). We conclude by relating the present results and perspective to both previous and subsequent studies. PMID- 11038613 TI - Initial frequency of alleles for resistance to Bacillus thuringiensis toxins in field populations of Heliothis virescens. AB - The risk of rapid pest adaptation to an insecticide is highly dependent on the initial frequency of resistance alleles in field populations. Because we have lacked empirical estimates of these frequencies, population-genetic models of resistance evolution have relied on a wide range of theoretical estimates. The recent commercialization of genetically engineered cotton that constitutively produces an insecticidal protein derived from the biocontrol agent, Bacillus thuringiensis (Bt) has raised concern that we lack data needed to quantify the risk of insect pests such as Heliothis virescens rapidly adapting to this ecologically valuable class of toxins. By individually mating over 2,000 male H. virescens moths collected in four states to females of a Bt toxin-resistant laboratory strain, and screening F1 and F2 offspring for tolerance of the toxic protein, we were able to directly estimate the field frequency of alleles for resistance as 1.5 x 10(-3). This high initial frequency underscores the need for caution in deploying transgenic cotton to control insect pests. Our single-pair mating technique greatly increases the efficiency of detecting recessive resistance alleles. Because alleles that decrease target site sensitivity to Bt toxins and other insecticides are often recessive, this technique could be useful in estimating resistance allele frequencies in other insects exposed to transgenic insecticidal crops or conventional insecticides. PMID- 11038614 TI - The capital-asset-pricing model and arbitrage pricing theory: a unification. AB - We present a model of a financial market in which naive diversification, based simply on portfolio size and obtained as a consequence of the law of large numbers, is distinguished from efficient diversification, based on mean-variance analysis. This distinction yields a valuation formula involving only the essential risk embodied in an asset's return, where the overall risk can be decomposed into a systematic and an unsystematic part, as in the arbitrage pricing theory; and the systematic component further decomposed into an essential and an inessential part, as in the capital-asset-pricing model. The two theories are thus unified, and their individual asset-pricing formulas shown to be equivalent to the pervasive economic principle of no arbitrage. The factors in the model are endogenously chosen by a procedure analogous to the Karhunen-Loeve expansion of continuous time stochastic processes; it has an optimality property justifying the use of a relatively small number of them to describe the underlying correlational structures. Our idealized limit model is based on a continuum of assets indexed by a hyperfinite Loeb measure space, and it is asymptotically implementable in a setting with a large but finite number of assets. Because the difficulties in the formulation of the law of large numbers with a standard continuum of random variables are well known, the model uncovers some basic phenomena not amenable to classical methods, and whose approximate counterparts are not already, or even readily, apparent in the asymptotic setting. PMID- 11038615 TI - Evolution of dendritic patterns during alloy solidification: onset of the initial instability. AB - The evolution of a dendritic pattern from a planar solid-liquid interface during directional solidification of a binary alloy was investigated experimentally. The model alloy used was the transparent organic crystal succinonitrile doped with the laser dye coumarin 152. The buildup of solute ahead of the initially stable planar interface and the subsequent instability of the planar front were measured in detail and compared with recent theoretical calculations by Warren and Langer [Warren, J. A. & Langer, J. S. (1993) Phys. Rev. E 47, 2702- 2712]. The fluorescence of coumarin 152 was used for direct observations of the evolution of the solute concentration profile ahead of the initially planar solid-liquid interface. UV absorption was used to produce thermal perturbations of the sample that generated spatially periodic modulations of the planar interface. This technique allows for measurement of both positive and negative linear growth coefficients (determined from the growth or decay rate of the modulation after the perturbation is switched off) for a large range of wave vectors. Measurements of the evolution of the concentration profile and the linear growth coefficients, and the occurrence of the initial instability, were in good agreement with the Warren-Langer predictions. PMID- 11038616 TI - Evolution of dendritic patterns during alloy solidification: From the initial instability to the steady state. AB - The evolution of the crystal-melt interface was investigated during directional solidification of a dilute binary alloy, starting at the marginal stability time t(i) at which the planar interface first becomes unstable. The time delay between t(i) and the crossover time t(0) at which the interface modulation becomes observable was determined experimentally. The interface morphology was analyzed as the cellular pattern appeared, and it was followed through the coarsening phase to the final steady-state dendritic pattern. The relevance of the initial instability for steady-state pattern selection was verified experimentally, and some aspects of the coarsening dynamics were measured and compared with theoretical predictions of Warren and Langer [Warren, J. A. & Langer, J. S. (1990) Phys. Rev. A 42, 3518-3525; Warren, J. A. & Langer, J. S. (1993) Phys. Rev. E 47, 2702-2712]. PMID- 11038617 TI - Tracking the motion of chemisorbed molecules on their adsorption sites. AB - A new method, the time of flight-electron-stimulated desorption ion angular distribution method (TOF-ESDIAD), is described. The method permits the mapping of the lateral momentum distribution of adsorbed species on their adsorption sites. Examples of the study of the rotation of chemisorbed PF3 molecules and the frustrated lateral translation of chemisorbed CO molecules, both on single crystal metal surfaces, are given. The observed anisotropy of the frustrated translation of CO is postulated to occur as a result of the 2pi* orbital alignment in chemisorbed CO along the close packed direction of the Cu(110) substrate. PMID- 11038618 TI - An extension of the steepest descent method for Riemann-Hilbert problems: the small dispersion limit of the Korteweg-de Vries (KdV) equation. AB - This paper extends the steepest descent method for Riemann-Hilbert problems introduced by Deift and Zhou in a critical new way. We present, in particular, an algorithm, to obtain the support of the Riemann-Hilbert problem for leading asymptotics. Applying this extended method to small dispersion KdV (Korteweg-de Vries) equation, we (i) recover the variational formulation of P. D. Lax and C. D. Levermore [(1979) Proc. Natl. Acad. Sci. USA76, 3602-3606] for the weak limit of the solution, (ii) derive, without using an ansatz, the hyperelliptic asymptotic solution of S. Venakides that describes the oscillations; and (iii) are now able to compute the phase shifts, integrating the modulation equations exactly. The procedure of this paper is a version of fully nonlinear geometrical optics for integrable systems. With some additional analysis the theory can provide rigorous error estimates between the solution and its computed asymptotic expression. PMID- 11038619 TI - Initial cash/asset ratio and asset prices: an experimental study. AB - A series of experiments, in which nine participants trade an asset over 15 periods, test the hypothesis that an initial imbalance of asset/cash will influence the trading price over an extended time. Participants know at the outset that the asset or "stock" pays a single dividend with fixed expectation value at the end of the 15th period. In experiments with a greater total value of cash at the start, the mean prices during the trading periods are higher, compared with those with greater amount of asset, with a high degree of statistical significance. The difference is most significant at the outset and gradually tapers near the end of the experiment. The results are very surprising from a rational expectations and classical game theory perspective, because the possession of a large amount of cash does not lead to a simple motivation for a trader to bid excessively on a financial instrument. The gradual erosion of the difference toward the end of trading, however, suggests that fundamental value is approached belatedly, offering some consolation to the rational expectations theory. It also suggests that there is a time scale on which an evolution toward fundamental value occurs. The experimental results are qualitatively compatible with the price dynamics predicted by a system of differential equations based on asset flow. The results have broad implications for the marketing of securities, particularly initial and secondary public offerings, government bonds, etc., where excess supply has been conjectured to suppress prices. PMID- 11038620 TI - Configurations for a proof of principle stellarator experiment. AB - One of the serious limitations of tokamaks as reactors is the occurrence of disruptions. Stellarators designed by advanced computational methods provide an attractive alternative for a major experiment in magnetic fusion research. Configurations with approximate two-dimensional magnetic symmetry have been found with high beta limits and good transport. Specifications are given for a compact stellarator with three field periods and 18 moderately twisted modular coils that has equilibrium with robust flux surfaces, a deep magnetic well assuring favorable stability, and adequate confinement of hot particles at reactor conditions. Fast computer codes with sufficient accuracy to resolve the mathematical problems of equilibrium, stability and transport that arise in the more complicated geometry of the stellarator have produced this breakthrough. The mathematical analysis of the methods used is presented. PMID- 11038622 TI - The UNOS Scientific Renal Transplant Registry. AB - Based upon data reported to the UNOS Scientific Renal Transplant Registry regarding transplants performed between 1994-1998, the one- and 3-year graft survival rates for 16,288 recipients of living donor kidneys were 93% and 86%, respectively, with a half-life of 17 years. Among those were 2,129 transplants from HLA-identical siblings with one- and 3-year graft survival rates of 96% and 93% and a 39-year half-life, 3,140 sibling donor grafts matched for one HLA haplotype with 94% and 87% one- and 3-year survival rates and a 16-year half-life and 2,071 transplants from living unrelated donors with 92% and 86% one- and 3 year graft survival rates and a 17-year half-life. The overall results of 35,289 cadaver donor kidney transplants were 87% and 76% graft survival at one- and 3 years with a 10-year half-life. There was a 13% difference in 3-year graft survival rates when recipients of kidneys from donors over or under age 55 were considered separately and the half-life was 11 years for younger donors and 6 years when the donor was older (p < 0.001). A total of 4,688 (14%) of cadaver kidney recipients received an HLA-matched transplant. Their graft survival rates were 89% and 83% at one and 3 years and their graft half-life was 16 years compared with 86% and 76% one- and 3-year graft survival and a 10-year half-life for recipients of HLA-mismatched kidneys (p < 0.001). The recipient's age affected both graft survival and the cause of graft loss. Recipients aged 19-45 had a 78% 3-year graft survival rate compared with 72% for recipients over age 60 or under 18 (p < 0.001). However, 65% of graft losses after the first year among older recipients were due to death with a functioning graft compared with 18% among 19-45-year olds. Acute rejections accounted for 16% of graft failures after the first year when the recipient was aged 6-18. Immune failures decreased with increasing recipient age. The recipient's race also influenced graft survival rates. Asian recipients of cadaver kidneys had the highest graft survival rates of 91% and 85% at one and 3 years with a half-life of 18 years. The result for Whites and Blacks were significantly lower (87-86% at one year and 78% and 70% at 3 years, respectively; p < 0.001). The graft half-life was 12 years for Whites and 7 years for Blacks. DGF and acute rejection episodes during the early posttransplant period reduced 3-year survival of cadaveric transplants by 20% and reduced graft half-lives by 2 years (rejections) or 4 years (DGF). When rejections occurred in recipients with DGF, 3-year graft survival was 64%. Induction therapy with anti-T-cell reagents did not affect graft survival rates among patients with DGF, but reduced the incidence of early rejections from 27 14%. Rejections that occurred within the first 6 months had a more pronounced effect on subsequent graft half-lives (11.6 years without and 7.6 years with; p < 0.01) and increased the proportion of kidneys that failed because of chronic rejection from 31-43% between 1-3 years. More than 50% of diabetics received a simultaneous pancreas kidney transplant during this period and the graft and patient survival rates were significantly higher for recipients of the SPK transplants. When deaths with a functioning graft were censored, however, the graft failure rates were not significantly different. The major causes of death among cadaver kidney transplant recipients were cardiovascular (26%) and infections (24%) during the first posttransplant year. Between 1-3 years, the percentage of deaths due to infection fell to 15% and malignancies accounted for 13% of patient deaths. PMID- 11038623 TI - Liver transplantation in the United States: a report from the UNOS Liver Transplant Registry. AB - Transplants and centers Between 1988 and 1998 the number of liver transplants performed in the US more than doubled from 1,713 to 4,487; the number of centers increased from 59 to 116. The number of living donor, segmental, and multiple organ transplants also increased over time. The rate of increase in the number of centers has slowed over the last few years. Outcomes. Survival among pediatric recipients. The one- and 7-year graft survival rates for pediatric recipients were 72% and 62%, respectively. The one- and 7-year patient survival rates were 85% and 79%. Patient survival did not decrease much after the first 2 years and graft survival stabilized after 4 years posttransplant. Some of the factors associated with increased odds of graft failure and patient death at 6 months posttransplant included having a previous transplant; being hospitalized, in the ICU, or on life support at the time of transplant; creatinine > 2 mg/dl; donor age and race/ethnicity; and transplant type. Factors associated with decreased odds of graft failure or patient death were recipient gender, recipient race/ethnicity, having a metabolic disease and receiving a living donor liver. Among grafts/recipients surviving the first 6 months after transplantation, recipient race/ethnicity, primary liver disease, having a previous transplant, donor age and race/ethnicity, and transplant type were associated with a greater relative risk of graft failure and mortality. Survival among adult recipients. The one- and 7-year graft survival rates among adult recipients were 77% and 57%, respectively. The one- and 7-year patient survival rates were 85% and 67%. Survival rates decreased steadily at all time points following transplantation. Some of the factors associated with increased odds of graft failure and mortality at 6 months after transplantation were recipient age and race/ethnicity; primary liver disease; having a previous transplant; being hospitalized, in the ICU, or on life support at the time of transplant; longer cold ischemia time; older donor age, race/ethnicity, or gender; and having a non-identical recipient/donor blood type match. Having cholestatic liver disease/cirrhosis and shorter cold ischemia times were associated with decreased odds of graft failure and mortality. Many of these characteristics also affected grafts and patients surviving the first 6 months, including recipient race/ethnicity, primary liver disease, previous transplant, and donor age. PMID- 11038624 TI - Worldwide thoracic organ transplantation: a report from the UNOS/ISHLT International Registry for Thoracic Organ Transplantation. AB - 1. The number of heart transplant operations performed in the US has increased by 51 procedures between 1997 (2,294) and 1998 (2,345). The number of lung transplants decreased by 67 in 1998 (862). 2. The most frequently reported indication for heart transplantation in the US is coronary artery disease (44.6%). For other thoracic transplants, the most frequently reported indications include other/unknown (43.9%) for double lung, emphysema/COPD (53.5%) for single lung and other/unknown (53.2%) for heart-lung. The most frequently reported diagnoses for thoracic transplantation outside the US include cardiomyopathy (50.5%) for heart, cystic fibrosis (32.0%) for double lung, idiopathic pulmonary fibrosis (32.7%) for single lung and congenital heart disease (24.7%) for heart lung. 3. US heart transplant recipients were predominately male (77%), between 50 64 years old (51.4%) and White (81.7%). In contrast, US lung transplant recipients are predominantly female (51.3%), between 50-64 years of age (44.7%) and White (89.7%). No meaningful variance from the US recipient demographic profile was noted for the non-US recipients during the same time period. 4. Pediatric recipients (< 18 years of age) received 10.9% of the reported heart transplants and 6.5% of reported lung transplants. 5. One-year survival for thoracic transplants performed in the US was 83.2% for heart, 70.6% for lung and 62.5%. Five-year survival for US thoracic transplants was 70% for heart and 49.1% for lung. 6. Long-term patient survival rates were: 22.3% at 18 years for heart, 20% at 9 years for lung and 25% at 12 years for heart-lung recipients. 7. The most important risk factor for mortality of US heart recipients at one month, one and 5 years after transplantation was receipt of a previous heart transplant. Significant short-term risk factors included donor age, recipient age and ischemic time. Substantial long-term risk factors include older donor age, donor race and recipient race. 8. The factors having the most significant impact on lung mortality at all time points were related to either the patient's medical condition (e.g., in the ICU prior to transplant, requiring mechanical ventilation) or diagnosis. 9. Mechanical ventilation and previous transplant had the largest impact on heart-lung mortality. 10. For heart and lung recipients, the major cause of hospitalization during the first posttransplant year was infection. PMID- 11038625 TI - Analyses of pancreas transplant outcomes for United States cases reported to the United Network for Organ Sharing (UNOS) and non-US cases reported to the International Pancreas Transplant Registry (IPTR). AB - As of September 1999, almost 13,000 pancreas transplants had been reported to the IPTR, > 9,000 in the US and > 3,000 outside the US. An era analysis of US cases from 1987 to 1997 showed a progressive improvement in outcome (p < 0.04), with pancreas transplant graft survival rates (GSRs) going from 74% to 85% at one year for SPK cases, from 56% to 75% for PAK cases, and from 50% to 69% for PTA cases. The improvements were due both to decreases in technical failure (TF) rates (overall from 16% to 8%) and immunological failure rates (going from 6% to 2% for SPK, from 23% to 7% for PAK, and from 35% to 9% for PTA cases). The proportion of recipients > 44 years old increased from 5% to 24%, and the improved outcomes encompassed the older patients as well. In patients > 44 years old, SPK pancreas GSRs at one year increased from 69% for 1987-89 cases to 79% for 1996-97 cases (p < 0.03). Pancreas GSRs were also similar for recipients reported to have Type I or Type II diabetes (at 1 year, 84% and 81%, respectively, for 1994-99 SPK transplants), the latter designated in 3% of the recipients. Contemporary pancreas transplant outcomes were calculated separately for 1996-99 US and non-US cases. US patient survival rates at one year were > or = 95% in each recipient category, with one year pancreas GSRs of 84% for SPK (n = 2,502), 76% for PAK (n = 404), and 72% for PTA (n = 176) (p = 0.0001). The immunological graft failure rates for 1996-99 US SPK, PAK and PTA cases were 2%, 6%, and 10% at one year (p = 0.001). There was a progressive increase in the use of ED (as opposed to BD) for duct management, up to nearly 60% of US pancreas transplants by 1998. Approximately 18% of SPK ED transplants had venous drainage via the portal system. Pancreas GSRs were not significantly different for 1996-99 ED (n = 1,170) and BD (n = 1,203) US SPK transplants (84% and 85%, respectively, at 1 year), nor was there any difference in pancreas GSRs for systemic (n = 437) versus portal (n = 194) venous drained ED SPK transplants (84% and 83%, respectively, at 1 year). Interestingly, kidney GSRs were significantly higher for ED versus BD US SPK cases, 93% versus 84% at one year (p = 0.003). Duct management did matter for solitary (PAK and PTA) pancreas transplants. PAK pancreas GSRs were 80% at one year for BD (n = 238) versus 68% for ED (n = 156) US transplants. PTA pancreas GSRs were 78% at one year for BD (n = 98) versus 63% for ED (n = 73) US transplants. However, BD transplants were associated with a 12% conversion rate to ED by 2 years after transplantation. Analyses of outcome by immunosuppression for US cases showed pancreas GSRs to be higher in SPK recipients given MMF (87% at 1 year) than in those who were not (76% at 1 year). For PAK and PTA recipients, those given anti-T cell for induction and TAC and MMF for maintenance immunosuppression had the highest GSRs: 86% and 83%, respectively, at one year for BD pancreas transplants; not significantly different from the pancreas GSR (87% at 1 year) in BD SPK recipients also given anti-T cell for induction and TAC and MMF for maintenance immunosuppression. Analyses of US pancreas transplant outcome according to HLA matching showed no effect at all in the SPK category, while for PAK and PTA transplants an effect was seen at the A and B loci, strongest at the B loci. Matching for at least one antigen at both loci was associated with one-year pancreas GSRs of 85% for PAK and 74% for PTA, versus 70% and 60%, respectively, at one year for those who were not matched for at least one antigen at both the A and B loci. In regard to non-US cases, the overwhelming majority were in the SPK category (n = 528 for 1996-99), with one-year pancreas GSR of 79%, not significantly different from US cases. Approximately 40% of non US SPK cases were ED (n = 204), and, as in the US, the pancreas GSRs were similar for ED and BD transplants in this category. (ABSTRACT TRUNCATED) PMID- 11038626 TI - The UNOS OPTN waiting list, 1988-1998. AB - 1. On October 31, 1999, there were 71,024 registrations on the combined UNOS waiting list. Of these, 64% were awaiting kidney transplantation and 20% were awaiting liver transplantation. 2. The majority of patients on the UNOS waiting list on October 31, 1999 were blood type O (52%), White (59%) and male (58%), and awaiting their first transplant (85.9%). 3. Median waiting times have increased steadily for nearly every organ since 1988, especially for liver, kidney, and lung registrants. 4. Median waiting times to transplant were longest for kidney registrants (938 days). The shortest waiting times for this cohort were experienced by intestine registrants (161 days). 5. Death rates per patients waiting at risk have been declining for most patients awaiting life-saving organs and have remained relatively stable for those awaiting for a kidney transplant. Deaths for intestinal patients have risen every year since the list was created (1993), but appears to be stabilizing. PMID- 11038627 TI - Organ donation in the United States: 1990-1998. AB - 1. There were 5,799 cadaver and 4,274 living donors recovered in 1998, 29% and 101% increases, respectively, over those recovered in 1990. 2. The number of cadaver donors aged 50 or older has increased from 16% of all donors in 1990 to 29% of all donors in 1998. 3. The typical cadaver donor in 1998 was a White male with ABO blood type O between the ages of 18-34. In 1998, a typical living donor was a White female with ABO blood type O between the ages of 35-49. 4. Between 1990 and 1998, the percentage of minority donors increased for cadaver donors (18% to 24%), and for living donors (24% to 27%). 5. The number of living donors who were either spouses or unrelated to the recipient increased from 5% in 1990 to 18% in 1998. 6. California (10.2%) was most often listed as the state of residence for cadaver donors, followed by Texas (7.3%) and Florida (7.0%). 7. Cadaver donors are recovered most often on Tuesdays (15.2%), followed by Wednesdays (14.6%) and Fridays (14.3%). 8. Living donors are recovered most often on Wednesdays (32.4%), followed by Tuesdays (27.5%) and Thursdays (21.3%). 9. Cadaver donors are recovered most often in August and May (8.8%), followed by July and October (8.7%). 10. Living donors are recovered most often in June (10.5%), followed by July (10.0%) and August (8.3%). 11. In 1998, there were 21.4 donors recovered per million population in the United States. PMID- 11038628 TI - Steady improvement in short-term graft survival of pediatric renal transplants: the NAPRTCS experience. AB - This report of pediatric renal transplantation covers the years 1987-1998. Since its inception in 1987, the NAPRTCS has collected data on 6,038 transplants performed in 5,516 patients provided by 73 renal centers across the country. FSGS, together with developmental lesions of dysplasia and obstructive uropathy, account for 40% of all transplants. There has been a steady increase in the use of LD donors among children with 54% of all transplants in 1996 and 1997 being live-related. About 72% of LD transplants are performed in Caucasian children, with African-American children unfortunately receiving a disproportionate percentage of CD kidneys. There has been a steady decline in the use of CD kidneys recovered from young individuals and a gradual decline in the number of transplants performed in young recipients (< 6 years old). Graft survival for LD recipients was 91%, 84% and 79% at one, 3 and 5 years, respectively, and the comparative figures for CD recipients were 81%, 72% and 64%, respectively. Acute and chronic rejections account for most of the graft losses, with chronic rejection accounting for more than 30%. There has been a steady improvement in one-year graft survival of CD recipients with the 1997-1998 cohort exhibiting an improvement of 16% over the 1987-1988 cohort. This improvement has been brought about by eliminating the use of infant donor kidneys, reducing the number of random transfusions and increasing the maintenance dose of cyclosporine. Posttransplant growth continues to be poor, with catch-up growth being exhibited only in children under age 6. PMID- 11038629 TI - Report from the National Transplantation Pregnancy Registry (NTPR): outcomes of pregnancy after transplantation. AB - Specific data on pregnancies following transplantation continue to accrue in the National Transplantation Pregnancy Registry (NTPR) in each type of organ recipient, with the following conclusions: 1. While the majority of kidney recipients appear to tolerate pregnancy well, a small percentage develops rejection, graft dysfunction and/or graft deterioration. Overall, there is a slight increase in the mean postpartum creatinine level when compared with the prepregnancy level, which has been noted in previous investigations by the NTPR. One neonatal death attributed to thrombotic cardiomyopathy was noted in a set of twins of a tacrolimus-based kidney recipient, but no other death has been noted in any of the additional reports among the recipients given newer immunosuppression regimens. Follow-up of offspring of these recipients is ongoing. 2. No structural malformations have been noted among offspring exposed to mycophenolate mofetil, but exposures are limited. (5 mothers, 29 fathers). 3. Female liver recipients with biopsy-proven acute rejection during pregnancy appear to be at greater risk for both poorer newborn outcomes and recurrent rejection episodes. In the setting of acute rejection diagnosed during pregnancy, close attention is warranted, anticipating that birthweight may be lower and that a substantial percentage of these female recipients may have recurrent rejection episodes. 4. Pancreas-kidney grafts can maintain normoglycemia throughout pregnancy. A high incidence of maternal hypertension, prematurity and low birthweight have been noted, so, as in other recipient groups, these are high risk pregnancies. Maternal pancreas and kidney function must be closely monitored. 5. No specific prepregnancy predictors of adverse outcomes have yet been identified among heart or lung recipients although none of the deaths among heart recipients in the NTPR database occurred within 2 years of delivery. When compared with other solid organ recipients, female lung recipients may face higher risks, particularly related to rejection. Whether prepregnancy factors can help to predict either heart or lung recipients at risk requires continued study. 6. No structural malformations or significant learning disabilities have been noted in follow-up of the offspring of CsA-treated females, the largest group of offspring followed to date with a mean age of 4-5 years. Continued surveillance of children will be essential to determine if effects become apparent as age related developmental delays or other problems in immune function or fertility later in life. 7. Newer regimens as well as new combinations of agents will continue to be studied. It is essential that non-viable as well as viable pregnancy outcomes be reported to the registry (i.e., recipients with pregnancies that result in spontaneous abortion or termination should be included for study). True estimates of non-viable outcomes have been difficult to assess. Additionally, inclusion of reports of pathologic evaluations at delivery hospitals will be helpful to determine whether spontaneous abortions are a result of lethal malformations related to immunosuppressive or other medication exposure. Safety of pregnancy for parent and child remain the primary goals of the NTPR. PMID- 11038630 TI - The National Marrow Donor Program: improving access to hematopoietic stem cell transplantation. AB - The National Marrow Donor Program has established a large and ethnically diverse registry of volunteer, unrelated stem cell donors. More than 9,000 patients world wide have received transplants from NMDP donors. In recent years, substantial resources have been invested to increase that portion of the NMDP donor file with complete HLA-A, -B and -DR typing. While this effort has improved the likelihood of patients identifying a suitable matched unrelated donor, it has also focussed more attention on issues surrounding donor availability. Several initiatives underway or planned are intended to improve donor availability and increase the overall success of the unrelated donor stem cell transplantation. PMID- 11038631 TI - Searching for an unrelated haemopoietic stem cell donor--a United Kingdom perspective. AB - The worldwide search for unrelated stem cell donors (now over 6 million) is one of the great success stories of international cooperation in the medical field. The initial search report from Bone Marrow Donors Worldwide estimates the chance of finding a suitably matched donor for a given patient. Registries whose donors are HLA-A, -B and -DR typed present the optimal prerequisite to identify a suitable donor for most patients. High-resolution matching HLA class I and class II alleles of the donor and recipient improves clinical outcome after unrelated donor transplantation. The clinical results of unrelated donor bone marrow transplantation are continually improving reflecting improvements in HLA matching, GvHD prophylaxis and transplantation in a favourable phase of disease. However, matching each HLA allele may or may not be critical for successful stem cell transplantation. Some degree of HLA mismatch ("permissible" mismatches) may be tolerated, especially in children. Cord blood banks provide a supplementary source of unrelated stem cell donors, in particular to patients from ethnic minority groups. PMID- 11038632 TI - Renal transplantation at the University of Michigan 1964 to 1999. AB - The Michigan Kidney Transplant Program has existed for 35 years. Outcomes have improved dramatically as the one-year survival of cadaver kidney grafts increased from 25% to 85-90%. Patient deaths in the first year are now uncommon. Indications for renal transplantation have been extended to infants, the elderly, diabetics and to patients with other significant health problems who would not have been candidates in the past. Chronic administration of large doses of corticosteroids is no longer necessary and the associated morbidity is largely avoided. Improvements in immunosuppression, especially the introduction of cyclosporine, account for much of this progress. With success has come increasing demand. Unfortunately, the gap between the number of available donor kidneys and the number of patients listed for a cadaver transplant continues to increase rather than diminish. Greater acceptance of volunteer donation, as has occurred in our own program, will help to reduce this shortage. If the past forecasts the future, we can anticipate extraordinary advances during the next 35 years. PMID- 11038633 TI - The LifeLink Foundation and cadaver kidney transplantation in Tampa. AB - 1. LifeLink Foundation, a not-for-profit organization, has been the driving force and absolutely essential entity for kidney and liver transplantation in Tampa providing all the components (patient, organs and clinicians) save for inpatient hospitalization. It also plays a big role in the heart transplant program. LifeLink has increased the kidney transplant rate from the first 1,000 done in 17 years to the second 1,000 in 7 years and is on a pace for the third 1,000 in 5 1/2 years. 2. Because of its innovative programs, cadaver donor procurement by the Tampa LifeLink OPO has been roughly double the national average for the past 10 years. Because of cadaver kidney availability the median wait time from activation on the wait list to transplantation over the past 5 years was 159 days. The recent transplant rate is 14.7-22.7% higher than the national average, dependent upon the parameter measured. Similar results are seen for Tampa patients awaiting heart and liver transplantation. 3. The overall outcome of 1,184 cadaver kidney transplants performed in the decade 1989-98 was similar to that reported from the UNOS database in this series of publications. a) One- and 2-year graft survival increased 2% per year over the decade with a recent one year graft survival rate of 96%. The overall T1/2 was 10 years. b) Our disastrous 1994 results were quickly reversed by a more intense pretransplant medical evaluation, the introduction of mycophenolate mofetil, more aggressive and earlier treatment of rejection episodes, and mandatory T- and B-cell flow cytometry crossmatching for all transplants. The incidence of rejection episodes decreased from 40 to 20%, and the first year immunological graft loss decreased from 5%, to 1.9%, to 0.8%, to 1.4% and 0% over the succeeding 4 years. 4. Individual factors affecting allograft survival were strikingly similar to national data, although all did not react statistical significance probably due to the smaller numbers. a) Primary and second grafts had similar survival rates (p = 0.97) whereas the third or subsequent graft survival was 7-32% poorer (p = 0.02). b) Black recipients had survival rates 10-13% lower than Caucasians and other races (p = 0.003). c) Patients with a peak PRA > 50 had survival values 4 13% poorer than those with < 50 PRA (p = 0.14). d) Patients with 2-4 HLA mismatches had graft survival rates 4-10% poorer than those with 0-1 mismatch (p = 0.12), whereas those with 5-6 mismatches had rates 6-17% poorer (p = 0.04). e) Although 22% of our transplants were to patients > 60 years of age, there was no difference (p = 0.81 to 0.90) in graft survival for the age groups 0-40, 41-60 and > 61. However, the proportion of grafts lost due to patient death compared with all allografts lost, was very different at 21% in the youngest group, 43% in those 41-60 years of age, and 63% in recipients > 61 years. 5. The rate of delayed graft function with imported kidneys was higher (27 vs. 16%, p = 0.006) but essentially the same as local kidneys with the same ischemia times. However, 41% of local kidneys were transplanted within 12 hours of procurement. Totally, 78% of local kidneys were transplanted within 18 hours (11% DGF rate) versus 79% of imports being transplanted at > 18 hours (32% DGF rate). Ischemia time, not the kidney source is the key issue since: a) There was no difference in overall graft survival of imported versus local kidneys (p = 0.95) nor in comparing local versus import kidneys with (p = 0.66) or without (p = 0.69) DGF. b) There was, however, a 11-17% overall poorer graft survival over 3 years in kidneys with DGF (p < 0.001) seen with both local (9-18% poorer, p = 0.0002) and imported (12-19% poorer, p = 0.008) kidneys. c) Kidneys displaying DGF came from older donors (40 vs. 34 years, p = 0.023) and had longer ischemia times (21 vs. 15 hours, p < 0.0005). 6. Dual kidney transplants were started in late 1996 with older or marginal donors to provide a better chance of success fo PMID- 11038634 TI - Kidney transplantation at the University of Miami. AB - Of the 1,679 renal allografts performed at the University of Miami between January 1, 1979 and October 31, 1999, 1,154 were from cadaver donors (CAD), 515 were from living-related donors (LRD), and 10 were from living-unrelated donors. The 3 ethnic groups: Black Caribbean-African-American, Hispanic, and others were almost equally represented among recipients. Recipient ages ranged between 1-83 years. In the CAD group, HLA matching was emphasized so that no patient received a kidney with less than one DR match, and for the entire series a mean of 2.59 of 6 HLA antigens were matched between donors and recipients. Overall actuarial 20 year patient and graft survival rates were 65.3% and 30.7%, respectively, with 69.2% patient and 38.5% graft survival rates for LRD, and 65.6% patient and 29.0% graft survival rates for CAD recipients. Several factors adversely affected long term graft outcome. African-Americans had an overall 20-year graft survival rate of 13.6% compared with 34% for non African-Americans (p < 0.001) (not dependent on patient survival). Diabetic patients had an overall 20-year graft survival rate of 13.5% versus 34.2% for non-diabetics (primarily dependent on patient survival). In the category of non African-American, non-diabetic patients under age 36 (n = 412), the 20-year patient survival rates in the LRD and CAD groups were 85.0% and 79.3%, respectively, and the graft survival rates were 55.7% and 46.5%, respectively. This differed markedly from the results for the entire series. PMID- 11038635 TI - Renal transplantation in Helsinki: influence on long-term survival of early posttransplant factors. AB - 1. The half-life of kidney grafts performed at Helsinki University Central Hospital has tripled from 6.5 years in 1980-84 to 17.1 years in the 1990's. 2. Delayed graft function constitutes a significant determinant of later kidney graft survival. Five-year graft survival was 81% for patients with early graft function and 71% for those with DGF. 3. High (> 350 mg/L) early (10 +/- 2 days after transplantation) cyclosporine trough levels were associated with inferior long-term graft survival. The 5-year graft survival rate was 74% compared with 86% for patients with trough levels between 250-349 mg/L. 4. Posttransplant conversion of the crossmatch test against the kidney donor, especially in association with DGF, indicates a group with significantly worse prognosis. Among 76 patients who developed antidonor antibody after transplantation, half experienced an acute rejection within 100 days and the 5-year graft survival rate was 68% compared with 83% for non-converters. PMID- 11038636 TI - CMV in kidney transplantation: a single center experience over 22 years. AB - Analysis of a historic renal transplant population for risks of developing CMV disease demonstrated a low mortality (0.2%) and morbidity. In our population of 1,959 patients, 411 (21%) developed subclinical CMV infection and 220 (11%) had CMV disease which was severe in 41 (2%). Important factors for infection were baseline immunosuppression, indicating that triple therapy with the proliferation inhibitors, azathioprine and MMF, had significantly higher infection numbers in comparison to dual, CsA-based immunosuppression. The cumulative dose of steroids correlated strongly with an increased number of CMV infections and disease, as did the addition of ALG/ATG or OKT3 for either steroid-resistant rejections or induction therapy. While CMV serology had an impact on infection in cases of seropositive donors to seronegative recipients, seropositive patients, in general, demonstrated increased infection rates most likely due to reactivation of the virus. Prophylaxis had no impact on the incidence of infection but reduced the severity. PMID- 11038637 TI - Renal transplantation in cyclosporine-treated recipients at the Singapore General Hospital. AB - 1. The 5-year patient and graft survival rates for live-donor renal transplants undergoing transplantation with CsA-based immunosuppression at the Singapore General Hospital from 1985-1999 were 97.0% and 91.6%, respectively. The 5-year patient and graft survival rates for cadaveric renal transplant recipients from the same institution were significantly lower at 91.6% and 79.4%, respectively. Long-term graft survival as described by half-lives (T1/2) were 28.6 years and 20.4 years for live-donor and cadaveric grafts, respectively. 2. Tissue matching had no impact on graft or patient survivals in either live-donor or cadaveric renal transplants. However, sensitisation was associated with significantly worse graft survival in cadaveric transplant recipients. 3. Primary cadaveric transplant recipients enjoyed significantly better graft survival rates than retransplanted recipients (5-year survival rates of 80.1% and 65.0%, respectively; p = 0.032). 4. Among cadaveric renal transplant recipients, a single acute rejection episode was associated with worse long-term graft survival (T1/2 of 14.6 years and 37.0 years for those with and without rejection, respectively, p = 0.001). Likewise, delayed graft function was associated with worse overall graft survival (5-year graft survival rates of 70.7% vs. 86.8% in patients with and without DGF, respectively, p < 0.001). 5. Among cadaveric transplant recipients with a functioning kidney at 5 years after transplantation, 61.4% had normal renal function with serum creatinine levels < 141 mumol/L while 33.9% had mild renal dysfunction with SCr of 141-250 mumol/L. 6. The high graft and patient survival rates in this Asian population as reported from this single centre study may be attributed to selection of younger patients without overt ischaemic heart disease for transplantation and to good patient compliance to CsA therapy. Thus, recipient selection and ensuring patient compliance to treatment are key strategies in optimizing the use of a scarce resource such as organ transplants. PMID- 11038638 TI - Pancreas transplantation at the University of Wisconsin. AB - Based on the University of Wisconsin experience with 653 cadaver pancreas transplant performed since 1985, we noted that: 1. The overall 5- and 10-year patient survival rates were 87% and 80%, respectively. The 5- and 10-year pancreas graft survival rates were 70% and 60%, respectively, and the 5- and 10 year kidney graft survival rates were 80% and 60%, respectively. 2. An immunosuppressive regimen including TAC and MMF in both solitary pancreas and SPK transplants was very effective in reducing the rate of acute rejection and improving graft survival. 3. Pancreas graft survival in recipients of solitary pancreas transplants was equivalent to that in SPK recipients in the TAC-MMF era. 4. Anti-IL2 receptor monoclonal antibodies were safe and effective in solitary pancreas and SPK transplants. 5. Excellent short-term graft survival can be achieved in solitary pancreas transplants using enteric drainage. PMID- 11038639 TI - Simultaneous pancreas-kidney transplantation at the Ohio State University Medical Center. AB - Simultaneous pancreas-kidney transplantation is becoming an accepted procedure for the treatment of Type I diabetic patients with end-stage renal disease. Its value in Type II diabetic patient remains to be evaluated. With the improvements in technical skills and immunosuppression medications, the procedure has become safer than those performed in the previous decade. However, the diabetic host is a high-risk individual. Careful evaluation and selection is prudent to keep excellent results with this scarce organ. As we have seen in this series, death is the number one cause of graft loss and better preoperative evaluation will continue to be of significant value to lower the mortality rate of SPK transplantation. We are moving to the use of marginal donors and marginal recipients very carefully and more data is needed to prove its safety and cost effectiveness. At our center, long-term patient and graft survival have been acceptable (86% and 73%, respectively, at 5 years). We will continue to use SPK transplantation as the procedure of choice for excellent candidates. We may need to change our philosophy to use living-related kidney transplants followed by a sequential pancreas transplant to overcome some of the shortage of pancreatic organs. The issue of primary enteric drainage has not been addressed in this report due to our success with bladder drainage. Despite the lower threshold for enteric conversion, about 10% of our patients are converted. PMID- 11038640 TI - Simultaneous pancreas-kidney transplantation at the University of Pittsburgh. AB - Analysis of the SPK program at the University of Pittsburgh has led to a number of observations: 1. Under tacrolimus-based immunosuppression, without antibody induction, it has been possible to achieve (a) One- and 3-year actuarial patient survival rates of 98% and 95% (b) One- and 3-year actuarial kidney survival rates of 95% and 87% (c) One- and 3-year actuarial pancreas survival rates of 86% and 80% 2. Steroid withdrawal has been achieved in over half of the successfully transplanted recipients, with excellent outcomes and a low rate (4.7%) of subsequent rejection. 3. Bone marrow augmentation has been associated with (a) less rejection (b) less pancreatic graft loss to rejection (c) an increased ability to withdraw steroids 4. Rejection has been associated with a rising serum lipase. 5. Renal allograft rejection in SPK patients with elevated serum lipase levels has been seen in the setting of normal renal function. 6. Enteric drainage has been associated with a reasonably low complication rate. 7. SPK transplantation is a successful therapeutic option in selected type I diabetics with end-stage renal disease. PMID- 11038641 TI - Liver transplantation at the University of Pennsylvania and the Children's Hospital of Philadelphia. AB - The liver transplant program at the University of Pennsylvania and the Children's Hospital of Philadelphia experienced healthy growth in its clinical activity in the past 5 years. Patterns of referral and patient evaluation were established, care of patients while waiting on the list or being followed after transplantation was streamlined. We are now achieving excellent outcomes while transplanting relatively sicker patients. Innovative surgical procedures are implemented resulting in more efficient utilization of cadaveric and living-donor liver grafts. The protocols that are used for patient care are more standard, yet flexible and accommodate recent advancement in transplantation immunobiology. This progress of the clinical program was enhanced by careful preservation of the academic mission of the institution, which encourages the liver transplant faculty to be involved in NIH-supported clinical and basic science research. PMID- 11038643 TI - Cardiac transplantation at the Cleveland Clinic Foundation. AB - The past 15 years has seen a significant evolution of heart transplant patient selection criteria, definition of suitable donors, immunosuppressive strategies, infection prophylaxis and treatment, and post-transplant patient surveillance. Primarily important has been broadening of the donor suitability definition and an evolution toward transplanting more ill and hemodynamically unstable patients. Despite "pushing the envelope" with both patient and donor selection and with transplanted patients generally being more ill, we believe our outcomes at the Cleveland Clinic Foundation are exemplary. The one- and 3-year survival rates for 265 heart transplants performed during 1996-1998 were 88% and 81%, respectively. Key to the success of our program has been close interdisciplinary working relationships and respect, broad and expert consultative support services, a desire to investigate clinical challenges, and dedication to excellence. Of additional importance is the realization that no matter how difficult we believe our difficulties are, we realize that those of our patients are vastly greater. PMID- 11038642 TI - Liver transplantation at the University of Chicago. AB - Since our 1995 report, improvements in patient survival after liver transplantation have widened indications for liver transplantation and led to a greater imbalance between donor supply and need. The organ shortage is the major barrier to liver transplantation at this time. Despite expanded donor criteria, there has been only a marginal increase in the number of liver transplants performed nationally. We have used several approaches to decrease the demand for organs in both adults and children. Our center was one of the first institutions to use reduced-size, living-donor, and split liver transplants routinely. The use of reduced-size liver transplants has decreased as the use of split liver transplantation has increased. Both split liver transplantation and living donor transplantation play an important role in caring for pediatric and adult patients with end-stage and fulminant liver disease. We have concentrated our recent efforts to optimizing the technical aspects of living donor transplantation in order to decrease the need for retransplantation and further organ use. These efforts have dramatically increased graft survival. We have also focused attention on treating patients prior to transplantation in an attempt to eventually abrogate the need for traditional transplantation in some disease processes. With the use of hepatocytes and liver assist devices, we have demonstrated that we can provide a level of metabolic support not achieved with traditional medical therapy for patients with fulminant hepatic failure. As further advances in these therapies are made over the next several years, a concerted effort to bridge patients to recovery will be made. As liver transplantation has become more standardized, it has opened the door to more challenges. We have used liver transplantation in combination with cardiac transplantation to care for selected patients with end stage disease of both organs. This has been remarkably successful for the 3 patients transplanted at the University of Chicago. The immunologic benefit of this combination appears to be a decreased incidence of cardiac rejection. We have standardized the technical components of this combined operation to allow for optimal organ function and patient survival. PMID- 11038644 TI - Cardiac transplantation in over 1000 patients: a single institution experience from Columbia University. AB - Since 1977, the cardiac transplantation program at Columbia has performed 1,137 heart transplant operations with a current one-year survival rate of approximately 90% and a 5-year survival rate of approximately 75% representing the largest single institution experience in North America. Over 2 decades of experience in the selection of donors and recipients has permitted us to expand eligibility limits and relax conventional exclusion criteria allowing us to transplant high-risk donors and medically complex recipients with excellent results. Recipient characteristics, rather than those of the donor, substantially impact outcome following OHT and use of extended donors will improve allocation of donor organs particularly with marginal recipients. During the 2-decade long evolution in our transplant experience, substantial improvements have been made in the areas of immunosuppression, treatment of rejection, and handling of sensitized recipients. Frequent causes of late mortality such as graft rejection, infection, malignancy, and TCAD, have been significantly diminished in the modern area of immune manipulation but remain major causes of death and barriers to long term survival. The single biggest impediment to growth in OHT is the shortage of donor organs. We have attempted to address this issue by identifying patients who may be better served with a bridging surgical procedure such as a mechanical assist device, or alternative procedures such as transmyocardial laser revascularization, high-risk reparative surgery, or myocardial volume reduction operations. Ongoing research interests at Columbia including LVADs, immunologic sensitization, xenotransplantation, and vasculogenesis offer the potential for continued growth for treatment of end-stage heart disease into the next millennium. PMID- 11038645 TI - Heart transplantation among 233 infants during the first six months of life: the Loma Linda experience. Loma Linda Pediatric Heart TransplantGroup. AB - Two hundred thirty-three heart transplantations were performed in infants during their first 6 months of life at Loma Linda University between November, 1985 and June, 1999. Survival has now exceeded 13 years. Nearly 70% of infants are expected to live at least 10 years. Those transplanted during the first 30 days of life have about a 15% survival advantage at 10 years. Scarcity of donors continues to limit the transplantation effort. While acute rejection is the most common cause of late mortality, posttransplant coronary artery disease (PTCAD) is the leading cause of graft loss affecting 22 recipients (9.5%). The majority of patients are asymptomatic prior to diagnosis of PTCAD and are either retransplanted or dead within 6 months. Retransplantation (9 of 11 retransplantations for PTCAD) has been highly successful, with 10 year actuarial survival of 91%. Posttransplant lymphoproliferative disease (PTLD) has been found in only 7 patients (3%), most commonly in lymph nodes. Causes of late mortality include acute rejection (n = 16), PTCAD (n = 9), infection (n = 7), PTLD (n = 2), chronic graft dysfunction (n = 2), arrhythmia (n = 1), recurrent pulmonary vein stenosis (n = 1), and other noncardiac causes (n = 4). Infant psychomotor development is mildly delayed although cognitive development is normal. School age children are performing at the level of their peers with average achievement and low average intelligence testing. Heart transplantation is durable therapy for newborns and infants with structurally incurable and end-stage myopathic heart disease. PMID- 11038646 TI - Thoracic organ transplantation at Papworth Hospital. AB - More than 1,200 patients have now undergone thoracic transplantation at Papworth Hospital and about 90 transplants are performed annually. Papworth remains one of the largest transplant units in the UK. Unique activities include a very large heart-lung transplant program: 247 patients have now undergone heart-lung transplants and 73 domino heart transplants have been performed. The 5-year survival rates are 71% for heart transplants, 48% for heart-lung and 41% for lung transplants, respectively. Chronic obliterative bronchiolitis remains an important limitation for heart-lung and lung transplant survival. PMID- 11038647 TI - Lung transplantation at Loyola University Medical Center. AB - The Loyola Lung Transplant Program shows a long record of offering transplants to suitable recipients, with good clinical results. The overall one-year survival rate was 84% for 53 lung transplant recipients in 1998-99. Our local perception on donor management appears to be successful at increasing donor organ availability. In addition, continuous evolution in posttransplant care and willingness to utilize newer immunosuppressive agents has reduced our incidence of acute rejection episodes to 23% during the past 2 years. Time will tell if there is also a measurable reduction in bronchiolitis obliterans syndrome. Finally, longitudinal research on QOL after lung transplantation continues to buoy our spirits based on patient acceptance and satisfaction with results. We continue to be strong advocates for transplantation and organ donation. PMID- 11038648 TI - Three decades of allogeneic bone marrow transplants at the Princess Margaret Hospital. AB - A total of 1,122 patients with various hemopoietic disorders were transplanted at the Princess Margaret Hospital since 1970. The majority suffered from acute or chronic myeloid leukemia. Improvements in support strategies permitted a gradual escalation of the upper age limit for transplant candidates and resulted in better survival. The overall survival at 10 years of all patients transplanted consecutively either before or after 1986 increased from 30 to 50%. This change was observed independent of other transplant related risk factors and is predominantly attributable to the use of cyclosporine and ganciclovir. An improvement of similar magnitude was seen for transplant recipients presenting with good risk features. The 10-year survival of patients with acute and chronic myeloid leukemia transplanted from a fully matched sibling donor in first complete remission or first chronic phase increased from 40 to approximately 70%. The quality of life of surviving patients may not return to normal but appears to improve with time after the transplant. PMID- 11038649 TI - Review of transplantation--1999. AB - 1999 was both an exciting and a frustrating year for organ transplantation. The exciting developments in the past year include the wider use of the anti interleukin-2 monoclonal antibodies, the introduction of sirolimus in the immunosuppression armamentarium and greater experimentation with immunosuppression regimens that spare corticosteroids and calcineurin inhibitors. The Holy Grail of transplantation, the clinical induction of tolerance, remains elusive although experimental studies will certainly propel clinical regimens in the new millennium. Xenotransplantation is not, as yet, a clinically applicable option. The organ shortage is as severe as it has ever been. Islet transplants have not replaced solid pancreas transplantation. Despite these unattained objectives, the past year has been notable for many achievements and will serve as a springboard for even more notable successes in the new millennium. PMID- 11038650 TI - The center effect: is bigger better? AB - Grouping US kidney transplant centers according to the number of transplants performed during the past 10 years into those that reported more than 1,000 transplants and all others showed: 1. Ten-year graft survival rates at larger centers were no more than 5% higher than those at other centers. 2. Graft survival rates were most similar between the 2 center groups when the donor was an HLA-identical sibling or when the recipient had chronic glomerulonephritis. 3. Larger centers had a slightly but noticeably higher graft survival rate when the patient had juvenile onset diabetes, when the donor was older than age 60 and when the donor was a spouse. 4. Most differences in graft survival rates between larger and all other centers were apparent only 2-3 years after transplantation, suggesting that the differences reflect long-term management of the patients or that an historical difference existed that has disappeared in recent years. PMID- 11038651 TI - Transplant risks. AB - 1. Changes in serum creatinine is a potentially useful predictor of chronic rejection. Patients with 2 10% increases in creatinine values in 3 consecutive years between 1-5 years had 4 times the risk of chronic rejection graft loss than patients with stable creatinine. 2. Formation of HLA antibody may correlate with graft rejection since losing a kidney increased the risk of broad sensitization 5 fold and losing multiple kidneys increased the risk ten-fold. 3. Sensitization increased the risk of acute and chronic rejection while pregnancies decreased the risk of acute and chronic rejection suggesting that pregnancy may result in "beneficial" sensitization. 4. HLA matching was the most potent factor decreasing the risk of acute rejection 2-fold and chronic rejection by 62%. 5. The incidence of acute and chronic rejection have both decreased significantly since 1994. PMID- 11038652 TI - The influence of donor age on kidney graft survival in the 1990s. AB - Based on analyses of the UNOS Registry data for cadaver kidney transplants performed between 1991-1999 we showed that: 1. 15-40 year old donor kidneys provided the best one-year graft survival rates. When donors were analyzed with recipients, younger (0-10) and older (70-90) donors and recipients (Table 2) had the lowest one-year graft success rates. 2. Chronic loss rate, the constant rate of graft loss between one and 5 years, showed younger donor kidneys had a significantly lower chronic loss rate compared with each older donor category. Apparently the younger donor kidneys have a resiliency and nephron reserve that provides better long-term function. However, they may have lower short-term (1 yr) graft survival rates, possibly due to their small size. 3. Black and White donor kidneys had similar one-year graft survival rates; however, in every age group, recipients of White donor kidneys had significantly better 5-year graft survival rates than Black donor kidneys. There was also a noticeably lower chronic loss rate among recipients of White than Black donor kidneys. 4. HLA matched White donor kidneys had better one- and 5-year graft survival rates and lower chronic loss rates than HLA-mismatched kidneys. The matching effect was lost when the donor age increased beyond age 40. PRA had an effect both at one and 5 years after transplantation. The chronic loss rate was similar with high and low PRA. Therefore, PRA had a relatively short-term effect. 5. Cold ischemia time had a modest effect after 35 hours both at one and 5 years. However, the chronic loss rate was unaffected by CIT, suggesting prolonged ischemia time had a relatively short-term effect. 6. More focused attention on sensitization and lowered CIT can both have a significant effect on short-term graft survival rates. However, both matching and younger donor organs provide the best opportunity for better long-term graft success rates. PMID- 11038654 TI - [Pathology of the pancreas. Analysis of 172 resection specimens]. AB - New sophisticated surgical methods enabled interventions on pancreas which kept remaining without attention of gastrointestinal surgeons for a long time because of previously unsolvable reasons. Pancreatic resection specimens enter now the biopsy diagnostics the correct interpretation of which needs a perfect technology of processing. Biopsy under CT monitoring, fibroscopical techniques combined with contrast investigation, biochemical, cytological and bioptic analyses of samples brought new diagnostic aspects into pancreatic disorders. The authors present a classification analysis based on their group of 172 resection specimens from 165 patients: tumor lesions are represented by 52 specimens, inflammatory lesions by 112 specimens, 8 cases are of another nature or are lacking in a clear diagnostic classification. Bioptic investigation can settle the diagnosis of very rare nosological entities which were distinguished before only in necroptic material (adult nesidioblastosis, Schwachman-Diamond syndrome associated with ductal carcinoma of the head of pancreas, and others). PMID- 11038653 TI - Determinants of long-term survival of adult kidney transplants: a 1999 UNOS update. AB - Half-lives presented in Figure 11 summarize our findings regarding the long-term determinants of adult renal transplantation. The Figure's groups were created by cross-classifying various levels of the more important/interesting determinants identified through the multifactor analysis (Tables 1 & 2). As indicated by the black bar, half of all 16-30 year-old cadaveric kidneys transplanted into Caucasian recipients who survive beyond one year after transplantation will function for 14 years (i.e., the half-life equals 14 years). Unfortunately, in Blacks and Caucasians receiving an older donor kidney, the renal graft half-life falls to 6-8 years. When a Caucasian patient receives a spouse's kidney for transplant, the half-life increases to 17 years; when a Caucasian patient receives a HLA-matched cadaveric kidney, the half-life increases to 19 years; and, when an Asian patient receives a 16-30 year-old cadaveric kidney, the half life jumps to 22 years. Finally, Caucasian patients receiving a HLA-identical sibling donor transplant enjoyed the greatest half-life of 26 years. PMID- 11038655 TI - [Structural correlations in splenic tissue in idiopathic thrombocytopenic purpura in adults]. AB - Immunologic activation of the splenic white pulp seemed to be an important feature of the structural changes in the 8 splenectomized patients with ITP. However, destruction of the platelets with CD 16+ monocytes was the most prominent change in the Billroth's cords of the splenic red pulp. Classical NK cells did not play an important role in platelets destruction. PMID- 11038656 TI - [Primary hepatosplenic T-cell (gamma delta) lymphoma: morphology and immunohistochemistry in 3 cases]. AB - We analyzed one autopsy case and two biopsy cases of primary (hepato-)splenic lymphoma, diagnosed in numerous trephine bone marrow, spleen and liver biopsies. It is a distinctive "new" type of a rare T-cell lymphoma characterized usually by rearrangement of gamma delta chains of T-cell receptor. Morphologically, the lymphoma is composed of a cytologically monotonous proliferation of small to medium sized lymphocytes, with diagnostically characteristic intrasinusoidal spread in the bone marrow, spleen and liver. The involvement of the lymph nodes is always absent. Immunohistochemically, the tumor cells expressed constant CD3 positivity and negativity for B- and myelomonocytic antigens, together with an inconstant coexpression of CD43 and CD45RO. In contrast to other and more common primary B-cell splenic lymphomas, its biological behaviour is more aggressive. PMID- 11038657 TI - [Papillary fibroelastoma of the endocardium]. AB - Six cases of papillary fibroelastoma of the endocardium were presented. All cases appeared as incidental findings at autopsy. Grossly, they were verrucous or tuft like growths less than 1 cm in size on cardiac valves. Histologically, they consisted of a fibrocollagenous stalk and multiple papillary fronds with typical zonal architecture: central dense hyaline core containing elastic fibers surrounded by a layer of myxoid matrix and covered by hyperplastic endothelial cells. At the surface of papillary fronds there were foci of fibrin deposition. Histogenesis of papillary fibroelastomas is unclear, they are considered variously as primary benign tumours, hamartomas or organized thrombi. PMID- 11038658 TI - [Malignant branchiogenic cyst--fact or fiction?]. AB - A case of a cystic metastasis of squamous cell carcinoma of palatine tonsil in a 54-year-old male is reported, clinically presenting as a branchiogenic cyst. The grade of differentiation of the squamous cell epithelium lining the cyst was very variable: in some areas it was extremely well differentiated, thus resembling common benign branchial cleft cyst; transition to dysplastic areas with features of carcinoma in situ was visible and finally, in one small focus, invasion of the epithelial structures into the lymphoid tissue was observed. Despite its metastatic nature, the tumor fulfilled histological criteria of a so called malignant branchiogenic cyst (branchiogenic carcinoma). The authors discuss the existence of primary malignant branchiogenic cyst and the criteria necessary for its diagnosis. PMID- 11038659 TI - [Recommended procedure for histopathologic processing of lymph node biopsy]. AB - A standard procedure of sampling, transportation, fixation and further processing for microscopical investigation of lymph node biopsies is presented. It points out the importance of a perfectly stained survey section and well done immunohistochemical reactions as well as of sense of the second opinion for precise histopathological diagnosis. PMID- 11038660 TI - A procedure supplement for tissue processing from percutaneous renal biopsies. AB - The division of a tissue cylinder from percutaneous renal biopsy into several unequal parts for routine biopsy, ultrastructural analysis, and immunohistochemistry may infrequently cause absence of glomeruli rendering the slides for immunohistochemical staining unsuitable. Instead of previously stained hematoxylin and eosin slide serving for checking up of the presence or absence of glomeruli defrayed by Health Care Insurance Companies the proposed additional control cryostat slide was stained by the Sudan Red stain. Such slides were fully accepted by the Insurance authorities and providing information of the presence and number of glomeruli have eliminated the need for a control hematoxylin and eosin stain. This saved time and expenses and at the same time brought information on the presence, degree and location of steatosis. PMID- 11038661 TI - Cytokeratins and lung carcinomas. AB - BACKGROUND: Recently, cytokeratins (CK) were studied as tumor markers for many carcinomas. In lung cancer they appeared to be useful in distinguishing primary from secondary tumors, in histological typing as well as in evaluating patient's prognosis. However, the results have yet to be conclusive. In this study, expression of CK7, CK10/13, CK18, CK19, CK20 was investigated in a group of 72 surgically resected specimens of lung including 31 adenocarcinomas, 30 squamous cell carcinomas and 11 neuroendocrine carcinomas. Cytokeratin immunophenotypes were analyzed in comparison to histological characteristics of tumors, TNM stages and patients survival. RESULTS: CK7, CK10/13 and CK18 can be used in distinguishing the lung adenocarcinomas from the lung squamous cell carcinomas: CK7(+), CK10/13(-), CK18(+) for adenocarcinomas; CK7(-), CK10/13(+), CK18(-) for squamous cell carcinomas. Relatively higher CK7 and CK18 immunostaining rates of the squamous cell carcinomas with high keratinization, with high percentage of dead cells and with late stages of disease suggested their prognostic significance but it was not confirmed when comparing different survival groups. Both adenocarcinomas and squamous cell carcinomas were stained strongly with antibody against CK19 (90.3% and 86.7% respectively) but much less with anti-CK20 antibody (9.7% and 3.3% respectively). In general, neuroendocrine tumors of the lung were non-reactive for these cytokeratins except CK18, among them all carcinoid tumors expressed CK18 abundantly. PMID- 11038662 TI - [Malignant myoepithelioma of the parotid gland: cytologic and histologic features]. AB - A case of malignant myoepithelioma of the parotid gland in a 34-year-old female is presented. In the fine-needle aspiration material, there was predominance of poorly cohesive polygonal cells with marked nuclear pleomorphism; no mitotic figures were observed. Focally, fragments of myxoid metachromatic intercellular material were also present. Histologically, the tumor was encepsulated, showing focal invasion of the capsule and tumor thrombi in the capsular vessels. The tumor was predominantly solid and myxoid, composed of cells with epithelioid features, marked anisonucleosis and a low mitotic activity. Immunohistochemically, the cells revealed positive staining for S-100 protein, vimentin, cytokeratins, glial acidic fibrillary protein and carcinoembryonic antigen; only several cell groups expressed smooth muscle actin and desmin; muscle specific actin was uniformly negative. In differential diagnosis, it was important to distinguish malignant myoepitelioma mainly from pleomorphic adenoma (mixed tumor), benign myoepithelioma, carcinoma ex pleomorphic adenoma and malignant melanoma. The criteria of malignancy in myoepithelial tumors are discussed. PMID- 11038663 TI - [Extraskeletal myxoid chondrosarcoma with neuroendocrine differentiation]. AB - We describe a case of extraskeletal myxoid chondrosarcoma with neuroendocrine differentiation. The tumor occurred in subcutaneous tissue of the right popliteal region in a 50-year-old man. It measured 5 cm in diameter, was well circumscribed, lobular and gelatinous, and lacked any necrosis or hemorrhage. Histologically, the tumor structure was a typical of extraskeletal myxoid chondrosarcoma. The lesion was lobulated and contained small to medium-sized chondroblast-like cells with ovoid hyperchromatic nuclei and without prominent nucleoli. The cells created cords and nests and showed focally a perivascular rosette-like arrangement. A few of the tumor cells were spindle shaped. The myxoid matrix was stained with alcian blue and this reaction was resistant to prior treatment with hyaluronidase. PAS-positive glycogen was found in the cytoplasm of some tumor cells. Immunohistochemically, the tumor cells were diffusely positive for neuron specific enolase, monoclonal synaptophysin and vimentin. Following antibodies gave negative results: desmin, actins, S-100 protein, pancytokeratin, epithelial membrane antigen, chromogranin A, neurofilament protein, myelinic basic protein, glial fibrillary acidic protein. The patient is well four years after the wide excision of tumor and radiotherapy. Neuroendocrine differentiation in extraskeletal myxoid chondrosarcoma was described at first by Chhieng et al. in 1998 (1). Our observation confirms this interesting finding. PMID- 11038664 TI - [Renal angiomyolipoma with a clear cell component]. AB - Case report of a 52-year-old woman with angiomyolipoma of the left kidney. The tumor had appearance of a typical angiomyolipoma predominantly composed of spindle cells, some epithelioid cells and few large hyalinized vessels. Adipose tissue was concentrated into small foci. Unusual presence of large cells with a clear fine granular cytoplasm closely resembled cells of the "sugar tumor" of the lung. Epithelioid cells and occasionally spindle cells were HMB45 positive. A minority of cells also coexpressed S100 protein. Clear cells were usually strongly positive for HMB45 too. Our findings supported consideration of a close relation between clear cell ("sugar") tumor of the lung and angiomyolipoma. PMID- 11038665 TI - [Autopsy and biopsy findings in disorders of mitochondrial beta-oxidation of fatty acids. Role of the pathologist in the diagnostic process]. AB - Basic problems of the group of hereditary mitochondrial beta oxidation (BOX) disorders are presented with evaluation of the role of pathologists in the diagnostic process. The disorders manifest themselves clinically as usual by recurrent Reye-like episodes (acute hepatopathy and encephalopathy) typically in low age levels. Integral part of the clinical picture is often a myopathic symptomatology which at some cases may display even persisting character. The findings at the tissue level are dominated mostly by steatosis of the organ set with a high beta oxidation level (liver, heart, skeletal muscle, kidney) and toxic effects of the intermediate metabolic products of the derranged beta oxidation process. So far eighteen enzyme defects have been described affecting either transport of fatty acids across the mitochondrial membranes or their oxidative degradation at various levels, pointing to an absolute dependence of the final diagnosis on biochemical analysis. Pathologist's conclusion in cases dying without diagnosis is limited to suspicion of a BOX disorder only. However, pathologists can contribute significantly to unraveling and specification of the underlying BOX defect by collecting adequate samples of body fluids and of unfixed organs. Nevertheless, the validity of these samples, even if widely recognized, is limited. The best approach is to provide samples enabling to perform biochemical evaluation of the whole BOX process in integral mitochondrias. This requirement is fulfilled solely by establishing fibroblast tissue culture post mortem. PMID- 11038666 TI - [Verification of identification of infectious agents by culture in routine gynecologic cytological screening]. AB - The original Bethesda classification system for reporting cervical/vaginal cytologic diagnoses has claimed besides oncologic evaluation also a statement on the presence of infectious agents. Their diagnosis should be followed by appropriate treatment. Based upon the comparison of careful bacterioscopic study in a series of 175 routine cervical smears with the results of microbiological, virological and mycological examinations the following pathogens might be according to the authors' opinion-diagnosed as highly possible and recommended for laboratory verification: cocci, Gardnerella vaginalis, Klebsiella, Acinetobacter, Morganella Morgani, Candida, Trichomonas vaginalis, Chlamydia trachomatis and human papilloma virus. PMID- 11038667 TI - [Age determination by detection of ratios of the D and L forms of aspartic acid]. AB - Paper concerns evaluation of results of age estimation based on the increase of D aspartic acid during aging. Samples of dentin from lower canine teeth were used as experimental material. In addition, analytical pitfalls and new possibilities of d-amino acids detection were discussed. PMID- 11038668 TI - [Sensitivity of GC-MS in the detection of benzodiazepines in the urine in the form of trimethylsilyl derivatives]. AB - A sensitive evidence of trace concentrations of benzodiazepines and their metabolites in urine can be enabled after special sample preparation including enzymatic hydrolysis, special solid phase extraction, silylation and following analysis by gas chromatography with mass spectrometry in electron impact mode. After optimalization of the procedure the extraction recovery values in the range 50-85% are achieved. The scale of retention times with basic mass spectral data are presented for the spectrum of silylated benzodiazepines which can overlap some gaps in the standard toxicological literature up to now available. PMID- 11038669 TI - [Eradication of contagious diseases]. AB - Based on analysis of eleven-year intense epidemiological intervention against smallpox, a number of findings and demands ensued which should be met by an infectious disease to be included into the programme of eradication or elimination. The author mentions several episodes from the programme of smallpox eradication in which he participated as a member of a WHO team. Part of the paper is a detailed explanation of the terms eradication and elimination. The main part of the article is a characteristic of infections where the global programme of eradication or elimination is underway. At present the eradication of poliomyelitis and dracunculiasis is completed and elimination of tetanus of neonates as well as leprosy, all by the year 2000. By 2010 measles, possibly German measles and mumps should be eradicated and possibly leprosy and Chagas' disease and onchocerciasis should be eliminated. Also for other infections such as lymphatic filariasis, trachoma and non-veneric treponematoses more remote terms are given or are not yet given. Depending on the decision of WHO on the programme of global eradication, under precisely defined conditions seven other infections may be included: cysticercosis (Taenia solium), diseases caused by Haemophilus influenzae b, viral hepatitis A, rotavirus enteritis, diphtheria, whooping cough and tuberculosis. In the case of viral hepatitis B only elimination is foreseen. PMID- 11038670 TI - [Parvovirus B19 antibodies in the blood in 17-34-year-old women]. AB - In order to assess the proportion of women in fertile age who lack antibodies against parvovirus B19 the authors assessed in 2 groups of sera (63 and 40 sera) taken from 17-34-year-old women antibodies against this virus by the ELISA method (Seiken). Virus-specific antibodies, class IgG were detected only in 67% of the examined subjects which indicates that in one third of the women there is a risk of transplacental infection of the foetus in case of infection with the parvovirus B19 during pregnancy. PMID- 11038671 TI - [Indicators of progression of HIV infection and their application to therapy]. AB - The authors evaluated in a group of 217 HIV positive patients the mutual relationship of the number of CD4+T lymphocytes and the level of the viral load of HIV RNA. Using correlation analysis evidence was provided of a not very marked negative correlation of the two indicators. As it was assumed that the relationship of the two parameters is influenced by the applied therapeutic procedures which reduce in particular the viral load, the two parameters were evaluated in relation to treatment. The closest relationship of the two investigated parameters was found in the group treated by monotherapy with zidovudine, followed by the group treated with a combination of two preparations (two nucleoside inhibitors of reverse transcriptase). In the group treated by three preparations (two nucleoside inhibitors of reverse transcriptase and a protease inhibitor) the relationship of the two parameters was least close. Investigation of the level of the viral load of HIV RNA and number of CD4+T lymphocytes is of major importance for the introduction of antiretrovirus treatment and selection of a suitable combination of antiretrovirus preparations. It makes it also possible to follow up the effectiveness of this treatment. PMID- 11038672 TI - [An epidemic of Trichophyton tonsurans trichophytosis in wrestlers (1994-1996)]. AB - During the period between September 1994 and August 1996 the authors detected an epidemic of trichophytia superficialis caused by Trichophyton tonsurans among wrestlers (age 10-19 years) of three sports clubs in southwestern Slovakia following their participation in international contests. Underrating of the initial symptoms of the disease, lack of adherence to the ban to engage in contests before complete cure, the relative resistance to topical and systemic treatment etc. led to the development of infection in 52 wrestlers (37% exposed ones). The authors recommend as one of the measures to prevent further import and spread of dermatophytoses measures prohibiting participation of infected sportsmen in any contests at home and abroad. PMID- 11038673 TI - [Transfer of resistance to 3rd generation cephalosporins and aztreonam in strains of Klebsiella pneumoniae producing extended spectrum beta-lactamases]. AB - The authors demonstrated the transferability of antibiotic resistance genes in nosocomial strains of Klebsiella pneumoniae, isolated from newborn children at the Paediatric University Hospital in Bratislava. Strains were resistant to cefotaxime, ceftazidime and aztreonam. The determinants of resistance (carbenicillin, cephaloridine, cefotaxime, ceftazidime and aztreonam) were transferred to recipient strains of Escherichia coli K-12 and Proteus mirabilis P 38. The transfer of resistance determinant from donor strains was demonstrated by the analysis of the resistance spectrum in transconjugant clones of recipient strains by the method of indirect selection. The ability of production of extended spectrum beta-lactamases (ESBL) was demonstrated by the double disc diffusion test. Synergy between clavulanate and cefotaxime, clavulanate and ceftazidime and/or clavulanate and aztreonam indicated production of ESBL by these strains. PMID- 11038674 TI - [Effect of ofloxacin, pefloxacin, and tobramycin on the hydrophobicity of Klebsiella pneumoniae]. AB - The effect of ofloxacin, pefloxacin and tobramycin at subinhibitory concentrations (1/4, 1/8 and 1/16 of the MICs) on surface hydrophobicity of the Klebsiella pneumoniae strain was studied. The antibiotics tested decreased cell surface hydrophobicity in a dose-dependent manner. The most significant reduction of surface hydrophobicity was found after treatment with antibiotics at 1/4 of their MICs. Surface hydrophobicity of K. pneumoniae after exposure to these concentrations was decreased to 44% (ofloxacin), 50.7% (pefloxacin) and 56.1% (tobramycin) compared with controls. PMID- 11038676 TI - [Increasing confidence and strength through retrospection] [In Process Citation] PMID- 11038675 TI - [The role of astroviruses in viral gastroenteritis in children]. AB - The submitted paper summarizes the first data on the incidence of astroviruses in specimens of faeces of children suffering from gastroenteritis in the Slovak Republic. The presence of the astrovirus antigen was revealed in 21 specimens (11.2%) of the investigated groups comprising 188 children. Astrovirus infection only was detected in 8 specimens (4.3%) and mixed infection with astroviruses and rotaviruses in 13 samples (6.9%), incl. six from 0-11-month-old infants. The results indicate the need to extend the spectrum of enteral pathogens elucidating diarrhoeal diseases by the diagnosis of astroviruses. PMID- 11038677 TI - [The high affinity uptake of glutamate in nervous system and its regulation by various factors]. AB - The high affinity uptake of glutamate in nervous system is sodium dependent and is mediated by glutamate transporter proteins located both in the nerve endings which release glutamate as a transmitter and in the plasma membrane of glial cells. Four subtypes (GLUT1, GLAST, EAAC1 and EAAT4) of glutamate transporters have been cloned. The localizations, distributions and pharmacological properties for each of these subtypes are unique from one another. The high affinity uptake of glutamate is regulated by various factors, such as arachidonic acid, nitric oxide, free oxygen radicals, protein kinases, cytokines and growth factors. The regulations are obviously meaningful from the standpoint of physiology and/or pathophysiology. PMID- 11038679 TI - [Molecular mechanisms underlying gating activity of voltage dependent ion channels]. AB - A model of two-channel and its activation and inactivation proposed by Hodgkin and Huxley in 1950s have been continuously confirmed by molecular biological and electrophysiological studies since 1980s. The activation of Na+ and K+ channels seems to be dependent on screw-like rotation outward of S4 segment which is highly conservative and very rich in positively charged amino acid residues. The inactivation of Na+ channels appears to be related to "hinged lids" like movement of intracellular linker between domain III and IV; type N-, C-, and P- of inactivation can be divided in K+ channels, which seems to occur mainly in terminal N-, C- and region P, respectively and the N-type inactivation seems to be related to "ball and chain" like movement of terminal N. PMID- 11038678 TI - [Ceramide pathway]. AB - Ceramide may be generated from many metabolism routes. As a novel second messenger, ceramide plays important roles in a variety of physiologic or pathologic events relating to cellular growth, proliferation, differentiation, apoptosis and injuries. It is expected that new insights into the ceramide pathway would promote our understanding of the complex mechanisms for signal transduction. PMID- 11038680 TI - [Advances in the research of opioid receptor and ion channels of dorsal root ganglion neurons]. AB - With the development of cDNA cloning technique in molecular biology, the research of opioids and opioid receptor as well as the mechanism of peripheral analgesia have made a great progress in the fields of receptor researches including molecular structure, morphology, molecular pharmacology, ion channel and intercellular signal transduction systems. The functional properties of mu, delta and kappa opioid receptor were determined by their structures. Depending on different proportional distribution in the primary sensory neurons, opioid receptor activation increases or decreases the potassium current and inhibits the calcium channel. There are different second message systems which are involved in the signal transduction between opioid receptors and ion channels. In addition, pharmacological evidence has also proved the existence of sub-types of opioid receptor, and the activation of these receptors can inhibit transmitters release from primary sensory neuron directly, which may be an important mechanism that opioid receptor has an analgesic effect in the peripheral nervous system. PMID- 11038681 TI - [Molecular mechanisms underlying the synaptic vesicle cycle]. AB - Synaptic vesicle cycle in nerve endings is a processes of multiple steps including docking, exocytosis, endocytosis and vesicle recruitment. It is mediated by protein-protein cascade reactions of vesicles, axoplasm and presynaptic membrane. The molecular model for the key steps of synaptic vesicle cycle is presented and can be used as the basis for furthering our knowledge about the activities of central nervous system. PMID- 11038682 TI - [Mediation of excitatory amino acids in synaptic transmission of spinal nociceptive information]. AB - NMDA and non-NMDA receptors exist extensively on the spinal dorsal horn neurons and are involved in mediating neurotransmission of spinal nociceptive information. NMDA receptor mainly mediates the neurotransmission of cutaneous nociceptive information, while non-NMDA receptor mainly contributes to the neurotransmission of muscular and visceral nociceptive information. One of the main reasons for this difference may be that the cutaneous and muscular nociceptive inputs induced more releases of aspartate and glutamate, respectively. There are co-operative interactions among the regulatory sites within NMDA receptor-channel complex in modulation of spinal nociception. The co operative interaction between excitatory amino acids and substance P in modulation of spinal nociceptive information may occur on both the body and the dendrites of the nociceptive neuron. PMID- 11038683 TI - [An animal model of non-hereditary Alzheimer's disease and its behavioral and pathologic changes]. AB - An animal model of non-hereditary AD was built by lesioning nucleus basalis of Meynert (nbM) and to investigate the behavioral alteration by Morris water maze, the pathological changes by special staining for senile plaques(SPs), enzymatic cytochemical staining for AChE, and immunocytochemical staining for beta amyloid protein (beta AP) and tau protein. Transmission electron microscopic observation has also been made. Results showed: (1) loss of learning and memory ability; (2) occurrence of necrotic granules in cytoplasm and inclusion in axon hillock; accumulation of necrotic cell, and formation of SPs under light microscope; (3) accumulation of microtubules and formation of inclusion, increase of lysosome and edema and vacuolation of neurons and neuroterminals under electron microscopy; (4) decrease of synaptic density; (5) sharp decrease of AChE (acetylcholinesterase) positive fibers and neurons shown by immunocytochemical staining; (6) overexpression of beta AP and tau protein. PMID- 11038684 TI - [The neuroethological basis of learning and memory decline in aged rats]. AB - The spatial reference and working memory abilities, GFAP-positive astrocytes and GABAergic interneurons were studied quantitatively in young, aged memory unimpaired and aged memory-impaired rats according to their learning and memory abilities in the Morris water maze. The results showed that the prominent behavioural features of the aged memory-impaired rat were a change in the searching pattern, a decrease in the spatial reference memory, no significant decrease in the spatial working memory was observed. The decrease of GABAergic neurons, and the increase of GFAP-positive astrocytes in the dentate gyrus of the aged memory-impaired rat were more obvious than the other two groups. The main abnormalities of morphologic indexes were closely correlated with the decline of learning and memory in the aged rat. PMID- 11038685 TI - [Hypoxia-inducible factor-1]. PMID- 11038686 TI - [Homocysteine: an independent risk factor for atherosclerosis]. PMID- 11038687 TI - [Signal transduction in the regulation of smooth muscle contraction]. PMID- 11038688 TI - [Engineering antibody: from laboratory to clinic]. PMID- 11038690 TI - [Progress in the studies on immunomodulatory effects of 5-HT]. PMID- 11038689 TI - [The research progress on 5-hydroxytryptamine transporter]. PMID- 11038691 TI - [Molecular structure and functional properties of AMPA and KA receptors]. PMID- 11038692 TI - [Ischemic (hypoxic) preconditioning: a general biological phenomenon and its clinical prospects]. PMID- 11038693 TI - [Biological research progress of carbonic anhydrase]. PMID- 11038694 TI - [Progress in anticancer mechanisms of soybean isoflavones]. PMID- 11038695 TI - [Telomerase and tumor]. PMID- 11038696 TI - [Annexin, a family of phospholipid-dependent calcium-binding proteins and its role in cell secretion]. PMID- 11038697 TI - [Beta-lactamase inhibitor: tazobactam]. PMID- 11038698 TI - [Immunological tolerance induction by intrathymic injection]. PMID- 11038699 TI - [My life as a neuroscientist]. PMID- 11038700 TI - [Neurosteroids]. AB - The term "neurosteroids" applies to those steroids that are synthesized in nervous system independent of supply by peripheral endocrine glands. The neurosteroids biosynthetic pathway, the mechanisms of neurosteroids in regulating neuronal activities and the neuromodulatory effects of neurosteroids, particularly the bimodal regulation of GABAA receptors, have been reported. PMID- 11038701 TI - [Integration of visual signals in the brain: mechanisms and functional significance of synchronous oscillation]. AB - How are the functions performed by one part of the nervous system integrated with those of others? One possible way is by synchronous oscillation. We have reviewed recent advances in visual system, where synchronous oscillations have been intensively observed and investigated. This article is concentrated on discussing theoretical reasoning, experimental evidence, possible mechanisms underlying the generation and the functional significance of visual synchronous oscillations. Predictions on several prosperous areas were also outlined. PMID- 11038702 TI - [Early T cell development in gene-targeted mutant mice]. AB - Early intrathymic T cell development is highly coordinated and controlled by the interaction of a variety of molecules. The development of gene targeting technology provided a valuable tool to study the function of these molecules in vivo. The analysis of TCR and CD3 gene targeted mice indicated that CD44-CD25+ is a control point in early T cell development. At this stage, the expression of pre TCR complex (composed of pre-TCR alpha, TCR beta and CD3) and/or its combination with unknown ligand provide signals for further progression beyond the CD44-CD25+ stage. The deficiency of any component in pre-TCR complex would lead to the blockage of T cell development at early stages. PMID- 11038703 TI - [Advances in the experimental study on the role of helper T cell subsets in the pathogenesis and the treatment of autoimmune diseases]. AB - In this paper the functions of helper T(Th) cell subsets, their cross regulation and agents affected their differentiation, as well as the role of Th subsets in the possible pathogenesis of autoimmune diseases were reviewed. Furthermore, a lot of opinions on autoimmune diseases therapy through restoration of Th subsets balance were mentioned. The available measurements to be used include up regulation of CD1d, administration of relative cytokines, administration of Freund adjuvant, administration of monoclonal antibodies against cytokines, cytokine receptors, CD40 ligand or B7 molecule, as well as CTLA or CTLAIg pathway and so on. PMID- 11038704 TI - [Modulation of potassium channel by arachidonic acid]. AB - Arachidonic acid is an important modulator for the inflammatory process. It can directly or indirectly regulate potassium channels which exist widely in the membrane of various types of cells. The direct modulation changes the conformation of potassium channel protein by the interaction between arachidonic acid and the protein, or through the interference with cell membranes around channels. The indirect influence of arachidonic acid upon channels results from its metabolites caused by cylcooxygenase and lipooxygenase, protein kinase C, and [Ca2+]i. All these aspects provide new potential targets for pharmacological investigations. PMID- 11038705 TI - [Influence of several immunoenhancers on sleep and their mechanism]. AB - The influence of three immunoenhancers on animal sleep and their mechanism was studied. Isoprinosine, transfer factor and muramyl dipeptide(MDP) could promote animal immune function, and prolong the sleeping time of mice and rabbits. Tumor necrosis factor (TNF), an immune cytokine, could prolong animal sleep; MDP could increase the synthesis and secretion of TNF from cultured mice astrocytes by promoting the expression of TNF alpha mRNA, resulting in an increase of TNF level in mice brains; monoclone antibody of TNF could partly antagonize the promoting effect of MDP on rabbits sleep. These results suggest that TNF is an important mediator in the influence of immune system on sleep. TNF could increase the turnover rate of 5-HT in mice brains, which may be one of the mechanisms of TNFs' promoting effect on sleep. PMID- 11038706 TI - [Experimental study of the effects of pituitary adenylate cyclase-activating polypeptide (PACAP) and its mechanism on the vascular cell components--the possible relationship between PACAP and atherosclerosis]. AB - By using cultured vascular endothelial cell (EC) and smooth muscle cell (SMC) as model, the morphological and functional effects of PACAP on EC and SMC in normal and hyper-lipid culture circumstances as well as its possible mechanism of action were studied. The results showed that (1) PACAP may partly counteract the morphologic injury of EC and SMC produced by high lipid. (2) PACAP significantly increased the production of anti-AS substances by EC. (3) PACAP inhibited the proliferation of SMC. (4) PACAP reduced significantly the production of lipid peroxide by EC and SMC in hyperlipid circumstances. The present study suggests that PACAP has cytoprotective effect on EC and SMC and thus it is possible that PACAP has an anti-AS effects. PMID- 11038707 TI - [FasL-Fas/APO-1 (CD95) system]. PMID- 11038708 TI - [Progresses in the study of inhibitor of differentiation (id) family]. PMID- 11038709 TI - [Differential gene expression and aging]. PMID- 11038710 TI - [A novel family of transcription factors: signal transducers and activators of transcription]. PMID- 11038711 TI - [Progress in the study of nuclear factor kappa B]. PMID- 11038712 TI - [Receptor internalization and nuclear translocation involved in intracellular signal transduction]. PMID- 11038713 TI - [Recent studies on taurine in the development and aging of the central nervous system]. PMID- 11038714 TI - [Effect of taurine on calcium of cardiomyocytes]. PMID- 11038715 TI - [Progress in spontaneous release of neurotransmitter at the neuromuscular junction]. PMID- 11038716 TI - [Cationic liposome and gene transfer]. PMID- 11038717 TI - [Molecular mechanism of retinol transport]. PMID- 11038718 TI - [Calcium-induced calcium release in cardiac myocytes]. PMID- 11038719 TI - [Biological actions of nerve growth factor in pain sensation]. PMID- 11038720 TI - [Interleukin-1 and ischemic brain damage]. PMID- 11038721 TI - [Cytochrome P450s and cancer]. PMID- 11038722 TI - [Nitric oxide: an important messenger molecule in the sphincter of Oddi]. PMID- 11038723 TI - [Contributing as a philosophy of life]. PMID- 11038724 TI - [AMPA receptor desensitization and fast excitatory synaptic transmission]. AB - AMPA(alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) receptor, an ionotropic glutamate receptor subtype, mediates fast excitatory synaptic transmission in central nervous system. Recently, the distinct characteristics of AMPA receptor desensitization have been gradually revealed and a number of chemicals which selectively modulate AMPA receptor desensitization have been developed. Many lines of physiological and pharmacological evidence show that AMPA receptor desensitization plays an important role in fast excitatory synaptic transmission by affecting transmission efficacy of individual synaptic bouton, integrative functions of neuron, and synapse plasticity. PMID- 11038725 TI - [Advancement on neurotransmitter transporters]. AB - The neurotransmitter transporter (NTT), a kind of glycoprotein situating on the presynaptic membrane, glial membrane, or vesicle membrane, has become the focusing point of neuroscience research in recent years. They could combine selectively with transmitters released into synaptic cleft and carry them back into cells to aid the termination of synaptic transmission. In this way, NTT took an important part in modulation of information between neurons. Study on NTT has not been reported domestically. This article reviewed recent studies on molecular structure, classification, study methods, distribution, function, modulating factors and gene modulation, focus of study and also unresolved problems of NTT. PMID- 11038726 TI - [Research progresses on signal transduction of neurotrophins]. AB - Binding of neurotrophins and their receptors lead to dimerization and autophosphorylation of trks. Activated trkA initiates the Ras pathway and finally opens the transcriptions of immediate early genes and delayed response genes or participates directly in physiological responses. Target-derived neurotrophins bind to and induce phosphorylation of trk receptors at the axonal terminal. Active trk or NT-trk or other signal molecules can be retrogradely transported along the axon to transduct messages to neuronal nucleus. There are local autocrine and paracrine mechanisms besides target-derived NTs. Following nervous system injury, increased gene expressions of NTs and their receptors and increased retrograde axonal transport are helpful to survive and regenerate for injured neurons. Lacking NTs and their receptors will result in serious abnormal development of nervous system of mice. PMID- 11038727 TI - [Antigen strategy: a new way for gene therapy]. AB - Synthetic purine-rich or pyridine-rich oligonucleotides have been shown to form stable triplex by binding to specific homopurine-homopyridine sequences in double stranded DNA. Several studies have suggested that triplex formation in the transcription-regulated regions of genes could inhibit their transcription in vitro or in vivo. This new technology was called antigene strategy, which would be of great value in the treatment of cancer and virus infection. However, the low stability of triplex and restricting target selection of homopurine homopyridine tracts in selected genes greatly reduced its utility in antigen strategies. Further work is still needed. PMID- 11038728 TI - [Myotrophic factors]. AB - A variety of substances have been found to show myotrophic effect and are named myotrophic factors. Myotrophic factors provided a new means to the research and treatment of motor neuron diseases and muscle degenerating diseases. In this article, we reviewed the studies of various myotrophic factors, and indicated the clinical significance and direction of their studies. PMID- 11038729 TI - [Structure and pharmacology of ryanodine receptors]. AB - Ryanodine receptors (RyR) exist at the membrane of the intracellular calcium stores and function as the calcium-release channels in the vertebrate animal cells. In the mammalian animals three isoforms of RyR, skeletal type (RyR1), cardiac type (RyR2) and brain type (RyR3), have been identified, and they are encoded by three distinct genes, ryr1, ryr2 and ryr3, respectively. Meanwhile, in the non-mammalian vertebrate animals other three isoforms of RyR have been found: alpha RyR and beta RyR present in the skeletal muscle simultaneously, whose amino acid sequences exhibit the high identity to that of the mammalian RyR1 and RyR3 respectively, and another distinct isoform of RyR in the avian heart. In addition, a large variety of cells of both mammalian and non-mammalian vertebrate animal expresses all or two of the three isoforms of RyR simultaneously. Molecular structure and pharmacology of these isoforms of RyR are reviewed in this article. PMID- 11038730 TI - [The ionic mechanism of hyperpolarization induced by glucocorticoid and its modulatory effects in mammalian neurons]. AB - This is the first report to investigate the rapid hyperpolarization by glucocorticoid in coeliac ganglion neurons of guinea pig in vitro. Hydrocortisone 21-hemisuccinate (F-suc) caused rapidly hyperpolarization in neurons; hyperpolarization persisted after the elimination of synaptic inputs and after suppression of protein synthesis with actinomycin D; F-suc also induced hyperpolarization when picrotoxin or Ca(2+)-free in perfusion solution; TEA and 4 AP can inhibit the hyperpolarization of F-suc. These results provided evidence that F-suc's hyperpolarization may be mediated by potassium channel in the membrane. Also F-suc suppressed the discharges of neurons; and modulated rapidly effects of neurons to excitatory or inhibitory amino acids (P < 0.05). PMID- 11038731 TI - [Enhancement of glycine-gated Cl- channel currents by 5-HT and NA in rat sacral dorsal commissural neurons is mediated by protein kinases]. AB - The effects of 5-HT and NA on glycine-gated Cl- channel currents(IGly) and its intracellular mechanisms were investigated in acutely dissociated rat sacral dorsal commissural neurons by using nystatin perforated patch clamp recording. It was found that: (1) the activation of 5-HT2 receptor coupled to IAP-insensitive G proteins increases intracellular DAG formation through the activation of phospholipase C(PLC). The accumulation of DAG increases the Ca(2+)-independent or novel PKC(nPKC) activity, resulting in the potentiation of IGly; (2) the activation of alpha 2-adrenoceptor by NA coupled to IAP-sensitive G-proteins reduces intracellular cAMP formation through the inhibition of adenylyl cyclase (AC). The reduction of cAMP decreases PKA activity, resulting in the potentiation of IGly. PMID- 11038732 TI - [Discovery of endomorphin is an important breakthrough in opioid research]. PMID- 11038733 TI - [Leukemia inhibitory factor and its physiological functions during embryo development and blastocyst implantation]. PMID- 11038734 TI - [Mechanism of regulation of Ca(2+)-sensitivity in smooth muscle contraction]. PMID- 11038735 TI - [Signal transduction of interleukin 12]. PMID- 11038736 TI - [Regulation of tumor necrosis factor production by monocytes/macrophages and the alteration in aging]. PMID- 11038737 TI - [FHIT: a tumor suppressor gene which links the fragile sites and tumors]. PMID- 11038738 TI - [Insulin-like growth factors and diabetic neuropathy]. PMID- 11038739 TI - [Research progress in long-term depression of synaptic transmission]. PMID- 11038740 TI - [Opposing effect signal transduction pathway and oxidative stress induced gene expression]. PMID- 11038742 TI - [Molecular mechanisms in temperature acclimation]. PMID- 11038741 TI - [Macrophages and wound healing]. PMID- 11038743 TI - [Effect of noradrenaline on brown adipose tissue recruitment]. PMID- 11038744 TI - [Telomerase: a new target for drug design and gene therapy]. PMID- 11038745 TI - [Role of protein phosphatase in the modulation of ion channel activity]. PMID- 11038746 TI - [Neurotic effect of neuropeptides]. PMID- 11038747 TI - [Progress in the research on imidazoline receptors]. PMID- 11038748 TI - [LBP/CD14 system and its relationship with endotoxin activities]. PMID- 11038749 TI - [Inhibition of estrogen receptor-positive human breast carcinoma cell growth by retinoic acid]. AB - OBJECTIVE: To evaluate whether the growth inhibition by retinoic acid and RAR alpha mRNA expression levels were affected by the change of ER expression. METHODS: The ER-negative breast cancer cell line MDA-MB-231 was transfected with the ER gene by stable transfection. RESULTS: In ER-transfected cells not only was the RAR alpha mRNA expression increased, but their growth was inhibited by retinoic acid as well. Estrogen could greatly stimulate the RAR alpha gene expression not only in established ER-positive cell lines but also in ER transfected MDA-MB-231 cells. CONCLUSION: Our data strongly suggest that ER mediated enhancement of RAR alpha levels play an important role in RA inhibition of human breast cancer cell growth. PMID- 11038750 TI - [Prediction and binding of a mutant p53 peptide to MHC class I molecule]. AB - OBJECTIVE: To predict tumor antigenic peptides from the intact amino acid sequence of tumor antigens, thereby providing a guide to the design and development of tumor peptide-based vaccines. METHODS: Antigenic peptides were predicted by using a computer program PEPMOTIF based on MHC class I allele specific consensus motifs; and a scoring system was established to assess the likelihood of the peptides to bind to MHC class I molecules. MHC class I-peptide binding assay and MHC class I-peptide stabilization assay were used to determine the binding ability and affinity of the predicted peptides. RESULTS: Three peptides derived from mouse mutant p53 were predicted. mp53P132F(LNKLFFQL) was shown to bind to H-2Kb, but not H-2Db. mp53P246S (MGGMNRSPIL) and mp53P270C (GRDSFEVCV) bound to neither H-2Kb nor H-2Db. A relationship was found between peptide scores and binding of the peptides to the relevant MHC class I molecules. CONCLUSION: The computer program PEPMOTIF combining with a scoring system is a simple and efficient tool for predicting MHC class I binding peptides derived from antigenic proteins. PMID- 11038751 TI - [Inhibition of in vivo growth of lung carcinoma cells after transfection with gap junction gene Cx43]. AB - OBJECTIVE: To explore the effects of expression of gap junction gene and cell cell communication on tumor growth. METHODS: A highly metastatic human lung carcinoma cell line PG was used. PG cells were defective of gap junctional intercellular communication(GJIC) and lacking expression of gap junction gene Cx43. By transfection, Cx43 cDNA was introduced into PG cells and blank vector cDNA was used as mock control. By using Northern-blot, dye-transfer methods and examinations of in vitro/in vivo growth, stable Cx43 transfectant cells were studied. RESULTS: The mock control cells resembled untransfected PG cells in lacking expression of Cx43 and GJIC function. They grew fast in soft agar(colony formation rate 11.6%) and in nude mice (average tumor weight 3.47 g in 28 days). The Cx43 transfectant cells showed increased level of Cx43 mRNA and increased function of GJIC. Cell growth in soft agar and in nude mice was markedly retarded. The inhibition rate was 90% and 75%, respectively. CONCLUSION: Increased expression of gap junction gene Cx43 induced tumor-suppressing effects in human lung carcinoma cells. PMID- 11038752 TI - [The expression of mdm2 gene related to the invasiveness of human hepatocellular carcinoma]. AB - OBJECTIVE: mdm2 gene was used as an index to study the molecular mechanism related to the invasiveness of human hepatocellular carcinoma (HCC). METHODS: The expression of mdm2 gene in 34 HCC samples and 19 paratumor liver tissues and the relationship between the expression of mdm2 gene and invasiveness of HCC were studied using reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: The expression level of mdm2 gene (x +/- Sx) in HCC samples was higher than that in paratumor liver tissues (50.18% +/- 5.24% vs. 27.70% +/- 7.43%, P < 0.05). The expression level of mdm2 gene in HCC samples with invasiveness was higher than that in HCC samples without invasiveness (59.09% +/- 7.28% Vs. 37.87% +/- 6.37%, P < 0.05). However, significant difference in expression of mdm2 gene was not found between small and large HCC as well as between HCC with and without capsule. CONCLUSION: The expression of mdm2 gene was related to the invasiveness of HCC. PMID- 11038753 TI - [Establishment of an apoptotic model induced by adriamycin in human hepatocellular carcinoma]. AB - OBJECTIVE: Establishment of cell apoptotic model of human hepatocellular carcinoma. METHODS: Human cultured hepatocellular carcinoma cell line HCC-9204 was exposed in vitro to different concentrations of adriamycin for various lengths of time. Cell apoptosis was ascertained by cell morphology under transmission electron-microscope, DNA electrophoresis and flow cytometry. RESULTS: When HCC-9204 cells were treated with adriamycin at 20 mumol/L for 3 hr, cell apoptosis occurred. Typical apoptotic bodies appeared and DNA ladder could be demonstrated on DNA electrophoresis. Apoptotic peak was also shown by flow cytometry. Apoptotic cells accounted for 72.1% of the cell population. Cells in G1, S and G2 phase of cell cycle were 75.9%, 24.1% and 0%, respectively. Only 35.5% of adriamycin treated cells remained alive. CONCLUSION: The apoptosis of human cultured hepatocellular carcinoma cells, HCC-9204 cell line, was successfully induced by adriamycin. Establishment of this model will help further explore the molecular mechanism of hepatoma cell apoptosis. PMID- 11038754 TI - [Inhibition of angiogenesis with antisense ODN of VEGF]. AB - OBJECTIVE: To investigate whether antisense oligodeoxynucletides(ODNs) of VEGF can become a kind of drugs for antitumor cancer treatment. METHODS: With a tumor model on the cornea of the mouse the inhibition of angiogenesis and tumor growth by 5 ODNs with different modifications and sequences was determined. RESULTS: ODNs with partial phosphorothioate modification, or with hairpin structure could effectively inhibit tumor-induced angiogenesis, but the linear ODN could not. The use of 2 ODNs at different sites gave the best results. CONCLUSION: VEGF antisense ODNs can hopefully become new drug with antitumor and antiangiogenesis activities. PMID- 11038755 TI - [Detection of Epstein-Barr virus in lymphoproliferative diseases by in situ hybridization]. AB - OBJECTIVE: To study the relationship between Epstein-Barr virus and various lymphoproliferative diseases in China. METHODS: Tissues from 214 patients with lymphoproliferative diseases were examined for the presence of Epstein-Barr virus (EBV) using in situ hybridization with EBV's LMP gene as probe. RESULTS: The positive rate of EBV in Hodgkin's disease(HD), Non-Hodgkin's Lymphoma(NHL), benign lymphoid proliferative disease(BLP) was 30.0%(15/50), 14.0%(18/129), 2.9%(1/35), respectively. Among NHL, EBV was detected in 28.1%(9/32) of high grade lymphoma (HNHL), 10.5%(9/84) of medium grade lymphoma(MNHL) and in none(0/9) of low grade lymphoma(LNHL). Significantly higher rates than that of BLP were observed in HD and HNHL. No significant differences in rates among patients with MNHL, LNHL and BLP. CONCLUSION: EBV may be involved in the pathogenesis of HD and HNHL, but not in MNHL and LNHL. PMID- 11038756 TI - [The growth-inhibiting effects of hyperthermic-hypoosmotic solution alone and in combination with chemotherapeutic agents on human gastric cancer xenograft in mice]. AB - OBJECTIVE: To evaluate the efficacy of hyperthermic-hypoosmotic solution alone and in combination with anti-tumor drugs in the growth inhibition of human gastric cancer xenografts in immuno-compromised Balb/c mice. METHODS: Cyclosporin A-treated mice bearing human gastric cancer Sy86B in the peritoneal cavity were treated i.p. twice with 43 degrees C double distilled water (DDW) or 43 degrees C DDW containing 0.05% chlorhexidine or 43 degrees C DDW with carboplatin(10 mg/kg). Normal saline at ambient temperature injected i.p. was used as control. RESULTS: Compared with the control, in mice treated with 43 degrees C alone or in combination with one of the two chemotherapeutic agents, significantly better therapeutic effects were obtained as shown by a decrease in tumor cell Brdu labeling index (BLI), proliferation index (PI) and DNA index (DI), prolongation of survival period and reduction of CEA and hEGF contents in the peritoneal effusion. The best treatment result was observed in mice receiving carboplatin in 43 degrees C DDW which was also superior to that in mice treated with carboplatin alone. CONCLUSION: Hyperthermic-hypoosmotic solution containing chemotherapeutic agent by i.p. infusion may be a treatment of choice in the management of peritoneal carcinomatosis in advanced stomach cancer patients. PMID- 11038757 TI - [The clinical course and treatment results of lung metastases from breast cancer]. AB - OBJECTIVE: To analyse the clinical course and treatment result of lung metastases from breast cancer. METHODS: 122 cases with lung metastases from breast cancer were treated by chemotherapy or chemotherapy plus endocrine therapy. Treatment results were assessed according to WHO criteria and survival rate estimated using the life table. RESULTS: The median time from initial treatment of primary tumour to lung metastases was 22 months. Sites of common consecutive metastases were lung, liver and bone. The overall response rate was 48% with a CR rate of 15%. Compared to non-DDP-encompassing regimen, the CR rate was higher in DDP-based chemotherapy (7% versus 21%, P < 0.05) with a longer median survival time (MST). The PR rate was higher in regimen containing anthracycline (48%) than in that without anthracycline (20%, P < 0.01). The response rate was similar between chemotherapy and chemotherapy plus endocrine therapy (P > 0.05). No difference in MST was observed between patients receiving anthracycline- and non-anthracycline encompassing regimens. The 1-,3-,5- and 10-year survival rate was 77%, 22%, 11% and 10%, respectively. The size of primary tumour, the length of disease-free interval, the number of lung metastases may provide additional information for predicting patients' survival after treatment of lung metastases. CONCLUSION: Combination chemotherapy, especially DDP-based chemotherapy may prolong survival time of patients with lung metastases from breast cancer. PMID- 11038758 TI - [Long-term results of preoperative chemotherapy for operable breast cancer]. AB - OBJECTIVE: To evaluate the long-term therapeutic results of preoperative chemotherapy for operable breast cancer. METHODS: Patients were divided into preoperative chemotherapy group (group A, 253 cases) and postoperative adjuvant chemotherapy group(group B, 284 cases). The group A patients received preoperative chemotherapy for 4 weeks, followed by radical operation two weeks after chemotherapy. Postoperative adjuvant chemotherapy began within 2 weeks after surgery, with the same chemotherapeutic regimen for 6 or more cycles in both groups. RESULTS: (1) The 5-year overall survival rate (OS) and disease-free survival rate(DFS) were 59% and 54.9%, respectively, for group A in stage III, which were higher than those of group B(28.3% and 20.8%, P < 0.05). (2) For group A in stage II and III, the 8-year OS were 81.4% and 46.9%, and DFS were 76.3% and 40.6%, respectively, which were higher than those of group B(OS: 67.4% and 20.7%, DFS: 62.9% and 13.3%, P < 0.05). (3) The 5-year and 8-year OS were higher for group A than those for group B in patients with T3, T4 or positive nodes > or = 4, (P < 0.05). CONCLUSION: The results show that preoperative chemotherapy can improve the short- and long-term survival of patients with operable, stage III breast cancer and long-term survival of patients with stage II breast cancer. PMID- 11038759 TI - [Prognosis of premenopausal patients with operable breast cancer related to the timing of operation]. AB - OBJECTIVE: Determining the fact if the timing of mastectomy during the menstrual cycle influences the prognosis of premenopausal women with breast cancer. METHODS: 67 cases of breast cancer women with regular menstruation were reviewed and analyzed retrospectively by medical statistics in terms of 5-year overall survival, 5-year relapse-free survival and 5-year local-regional relapse survival. RESULTS: Even though the 5-year overall survival and 5-year local regional relapse survival were not different statistically, timing of operation did have a tendency to influence outcome. Meanwhile, estrogen receptor played the most important role in influencing the above-mentioned aspects in comparison with other factors. CONCLUSION: We believe that operation during the last menstrual period might have influence on long-term survival of premenopausal breast cancer patients, but prospective studies are needed. PMID- 11038760 TI - [Comprehensive treatment of bone metastases of breast cancer: a clinical analysis of 70 cases]. AB - OBJECTIVE: Bone metastasis is common in breast cancer patients and its main symptom is bone pain. A series of methods were tried to relieve bone pain and to improve quality of life. METHODS: 70 cases of bone metastases of breast cancer were divided into 2 groups and treated with either chemotherapy, radiotherapy, endocrine therapy or isotope therapy alone or in combination. RESULTS: Endocrine therapy or isotope therapy given alone was more responsive than chemotherapy or radiotherapy alone. When the treatment modalities were used comprehensively, the response rate was 90% as compared to 56.4% in patients given single modality treatment (P < 0.01). CONCLUSION: Endocrine or isotope therapy given alone is more effective than chemotherapy or radiotherapy alone. Patients positive for estrogen and progesterone receptors, are more suitable for endocrine therapy. Comprehensive treatment is recommended. PMID- 11038761 TI - [Immunohistochemical study on the expression of c-erbB2 oncoprotein in breast tumors]. AB - OBJECTIVE: To examine the relationship between oncoprotein c-erbB2 over expression and prognosis of primary breast tumor patients, and find if it can be used as a prognostic indicator. METHODS: Eighty-five cases of paraffin-embedded specimens were used for immunohistochemical study. RESULTS: Over-expression of c erbB2 was observed in none of the 7 benign breast tumors. Twenty-three out of 78 cases of breast cancer showed over-expression with a positive rate of 29.4%. Over expressed c-erbB2 oncoprotein was not detected in 80% of patients with negative lymph node. In clinical stage III patients, 71% of them over-expressed c-erbB2. The expression of c-erbB2 and estrogen receptor(ER) was inversely correlated in 67% of breast cancer patients. Oncoprotein c-erbB2 over-expression had no correlation with lymph node status(P > 0.05), but was closely correlated to clinical staging(P < 0.05). CONCLUSION: Patients with over-expression of c-erbB2 oncoprotein showed poor prognosis. The c-erbB2 may be used for predicting the prognosis of breast cancer patients. PMID- 11038762 TI - [Pathological study of en bloc breast sections in papillomatosis and its malignant transformation]. AB - OBJECTIVE: To investigate the characteristics of breast papillomatosis, its malignant transformation and relation with main ductal papilloma. METHODS: A histopathological study by en bloc breast subserial sections was performed on 14 cases of highly proliferative papillomatosis and 22 cases of carcinoma arising from the highly proliferative papillomatosis. RESULTS: The two types of lesions were widely scattered over the sections, covering more than two quadrants in 71.4% (10/14) and 63.3% (14/22) of the breast, respectively. Crisscross and histological transition of the two lesions were observed frequently in the sections of carcinoma arising from the highly proliferative papillomatosis. These two kinds of lesion often coexisted with main ductal papilloma. CONCLUSION: Papillomatosis is a breast precancerous lesion and it may have similar pathogenetic mechanism as main ductal papilloma. PMID- 11038763 TI - [Serum tumor necrosis factor-alpha levels in non-Hodgkin's lymphoma patients and its relationship to prognosis]. AB - OBJECTIVE: To explore the relationship of tumor necrosis factor-alpha (TNF alpha) with non-Hodgkin's lymphoma (NHL) and to evaluate clinical implication of serum TNF alpha. METHODS: Serum TNF alpha levals were measured by double-antibody sandwich enzyme-linked immunosorbent assay (ELISA) in 76 NHL patients. Survival analysis was performed. RESULTS: Serum TNF alpha was increased in 55% newly diagnosed patients, and dropped to normal level on remission but increased again on relapse. Elevated TNF alpha levels of patients at diagnosis were significantly associated with Ann Arbor stage, bulky tumor and B symptoms, but weren't correlated to serum lactic-dehydrogenase (LDH) nor to beta 2-microglobulin (beta 2-MG). The survival period of patients with elevated TNF alpha levels was shorter than that of patients with normal TNF alpha levels. CONCLUSION: Elevated serum TNF alpha level is associated with high tumor burden and can be used as a prognostic marker. PMID- 11038764 TI - [Diagnosis and treatment of primary malignant tumors of the small bowel]. AB - OBJECTIVE: To study the clinical characteristics, effective diagnostic methods and therapeutic measures of primary malignant tumors of the small bowel so as to improve therapeutic effect. METHODS: In this paper 75 cases of primary malignant tumors of the small bowel were analyzed. Survival rates were calculated with Life Table, and computer Cox multivariate analysis model was used for survival analysis. RESULTS: The most frequent clinical manifestations of primary malignant tumors of the small bowel were abdominal pain, abdominal mass, emaciation and intestinal obstruction. Gastrointestinal barium meal roentgenography was performed in 13 cases, with an accurate diagnostic rate of 84.6%. The 1-,3- and 5 year survival rate after radical resection was 87.5%, 68.7% and 48.1%, respectively, while that for palliative resection was 57.9%, 33.8% and 24.1%, respectively. The factors including age, histological type, tumor site and type of resection were selected into Cox model in survival analysis. CONCLUSION: For primary malignant tumors of the small bowel, gastrointestinal barium meal roentgenography is the most valuable method for diagnosis, and surgical radical resection is the most effective therapy. The significant prognostic factors are age, histological type, tumor site and type of resection. Chemotherapy had no effect on prognosis. PMID- 11038765 TI - [Preoperative intraperitoneal versus intravenous carboplatin chemotherapy for advanced gastric cancer, pharmacokinetics and drug accumulation study]. AB - OBJECTIVE: To examine the accumulation of anticancer drugs in tumor tissues by different routes of administration. METHODS: Pharmacokinetic comparison of preoperative intraperitoneal (i.p.) and intravenous (i.v.) carboplatin chemotherapy was done in 10 patients each with advanced, resectable gastric cancer. Preoperatively, 300 mg/m2 carboplatin in 750 ml 0.9% sodium chloride was administered by i.p. or i.v. At 160-180 minutes after drug administration, peritoneal fluid, portal vein blood and peripheral blood were taken. At 240-270 minutes later, cancer tissues, peritumor normal tissues, omentum majus, peritoneum and cancer-free lymph nodes were collected during operation. Total carboplatin concentrations were measured by high performance liquid chromatography (HPLC). RESULTS: High concentrations of carboplatin were found in all tissues by i.p. administration. Drug concentration was highest in the peritonaeum, being 4 times as high as that after i.v. administration. Cancer tissue had higher drug concentration than did peritumor normal tissues. Compared to those after i.v. administration, concentrations in the peritoneal fluid, portal vein blood and peripheral blood were 13, 3 and 1.5 times as high, respectively. No significant differences were observed in drug concentrations in the peritoneum, omentum majus and lymph nodes after i.v. administration. CONCLUSION: For gastric cancer, carboplatin chemotherapy by i.p. administration is a better route for clinical practice. PMID- 11038766 TI - [Metastases to the mediastinal lymph nodes in lung cancer and their extensive dissection]. AB - OBJECTIVE: To study the metastatic frequency, distribution and feature of mediastinal nodes in lung cancer and to establish criteria for nodal dissection. METHODS: Clinical data of 386 patients who underwent resection of lung cancers in the past 9 years were analyzed retrospectively. Based on the distribution map of mediastinal lymph nodes developed by Naruke, ipsilateral hilar and mediastinal nodes were resected. RESULTS: 147 cases (38.1%) had mediastinal nodal involvement (N2). A total of 289 groups of nodes were dissected. The rate of N2 disease among squamous-cell, adeno-, small-cell and large-cell carcinomas was 30.1%, 44.1%, 48.0% and 50.0%, respectively. In 71 cases with N2 disease in the upper lobes, 146 groups of N2 nodes were resected, including 124 groups in the upper mediastinum (84.9%) and 22 groups in the lower mediastinum (15.1%). In 76 cases with N2 nodes in the lower and middle lobes, 143 groups of N2 nodes were dissected, including 76 groups in the upper mediastinum (53.1%) and 67 groups in the lower mediastinum (46.9%). Saltatory metastases occurred in 79 patients, accounting for 53.7% of N2 nodes and in 16 cases that occurred in the mediastinum (10.9%). CONCLUSION: The feature of mediastinal lymph node metastases in lung cancer may be saltatory and multiple. Cure can be achieved only after extensive dissection of the upper and lower mediastinal nodes. PMID- 11038767 TI - [Clinical significance of p21H-ras expression and H-ras codon 12 mutation in squamous intraepithelial lesion (SIL) and carcinoma of uterine cervix]. AB - OBJECTIVE: To study the clinical significance of p21 H-ras expression and H-ras codon 12 mutation in SIL and cervical carcinoma. METHODS: p21 H-ras expression and H-ras codon 12 mutation was detected in the same paraffin embedded tissues of 171 cases of cervical carcinoma, 68 cases of SIL, and 29 cases of chronic cervicitis, by using immunohistochemical and PCR-RFLP techniques. RESULTS: (1)p21 H-ras was over-expressed in 25.0% of the cases with low-grade SIL but in most of them the immunohistochemical staining was not strong (score < 3). p21 H-ras over expression was present in 64. 6% of the cases with high-grade SIL and in 35.4% of them, the staining was strong (score = 3). Even higher frequency of p21 H-ras over-expression was seen in cases with cervical carcinoma (66.1%) and in about one-half (49.7%) the staining was strong. (2) H-ras codon 12 mutation was only detected in stages II and III cervical carcinoma, with frequency rate of 27.0% and 52.5% respectively. The five-year survival rate of patients with H-ras codon 12 mutation (20.3%) was significantly lower than that without mutation (79.7%). Besides, there was a correlation between lymph node metastasis and H-ras codon 12 mutation. CONCLUSION: p21 H-ras expression is helpful for early detection of cervical carcinoma. Aggressive biological behavior of cervical carcinoma is significantly increased once the H-ras codon 12 mutation occurs. H-ras codon 12 mutation is helpful to judge prognosis of cervical carcinoma. PMID- 11038768 TI - [A primary study of the association of human leucocyte antigen and osteosarcoma]. AB - OBJECTIVE: To study the association of human leucocyte antigen and osteosarcoma in Chinese Han nationality. METHODS: The frequencies of HLA-A,B,DR,DQ locus antigens were tested in a group of 25 osteosarcoma patients and in a control group of 250 healthy persons by using complement-dependent microlymphocytoxity technique. The members of both groups were of Chinese Han nationality. Their results were compared statistically. RESULTS: The frequency of HLA-B35 was 0.400 in patient group and 0.048 in the control group. The relative risk of suffering from osteosarcoma in persons carrying HLA-B35 was 13.220 times as high as that in those without this antigen (P < 0.01). Patients with HLA-B13 had 12.048 fold increase in relative risk with poor prognosis compared to those without this antigen (P < 0.05). A tendency of worst prognosis was seen in patients who carried both HLA-B13 and HLA-B35, but patients with HLA-B40 had 7.057 times better relative safety for good prognosis than those without that antigen (P < 0.05). CONCLUSION: HLA-B35 is in close linkage to osteosarcoma susceptible gene in Chinese Han nationality. HLA-B13 and HLA-B40 may be separately related to the malignant and resistant genes of osteosarcoma. PMID- 11038769 TI - [Giant-cell tumor of bone: a review of diagnosis and treatment of 105 cases]. AB - OBJECTIVE: To summarize the experience in the diagnosis and treatment of giant cell tumor of bone, the selection of various operation methods and their effects were evaluated and discussed. METHODS: From 105 cases of giant-cell tumor of bone data were collected and reviewed. The clinical features were analyzed in association with radiological and histological characteristics. Prognosis was assessed by long-term follow-up. RESULTS: Among 105 cases, one had multiple giant cell tumors and two had distant metastases. In some cases, the radiological and histological appearances didn't accord with the clinical findings. The recurrence rate was 36% in patients treated with curettage. Although no recurrence was seen in patients treated by local excision, the joint function could not be fully retained. CONCLUSION: Diagnosis should be made on the basis of integrated clinical, radiological and pathological manifestations. Early diagnosis and operation are important. The function of joint must be retained as much as possible. Pre-operative embolization of tumor of sacrum and pelvis is very helpful to ensure operative success and to minimize recurrence. PMID- 11038770 TI - [Development of medical molecular biology]. PMID- 11038771 TI - [Some aspects of current research on medical molecular biology in China]. PMID- 11038772 TI - [Conference on the combination of molecular biology and clinical studies]. PMID- 11038773 TI - [Direct surgical clipping the carotid-ophthalmic aneurysms through a extradural intradural approach]. AB - OBJECTIVES: To exlore the approach for treating carotid-ophthalmic aneurysms and introduce experience in reducing postoperative complications. METHODS: We treated 31 patients with carotid-ophthalmic aneurysms, which accounted for 8.3% of all intracranial aneurysms we encountered in 1994 and 1997, by direct surgical clipping. RESULTS: No operative death occurred and the rate of postoperative complications was 19%. All aneurysms were occluded and later confirmed by angiography. CONCLUSION: The direct clipping of aneurysms involving the carotid ophthalmic artery is best treated by the extradural-intradural approach. PMID- 11038774 TI - [The impact of the missense mutation-ser20gly in islet amyloid polypeptide gene on NIDDM in Chinese]. AB - OBJECTIVES: To ascertain the presence of the Ser 20Gly mutation of islet amyloid polypeptide (IAPP) gene and its impact on NIDDM in Chinese. METHODS: In 896 Chinese, 825 were unrelated subjects (NIDDM in 609 and non-diabetics, 216) and 71 were family members of the pedigrees with IAPP gene-Ser 20Gly carrier probands detected from population screening. The mutation was examined by PCR-RFLP MspI digestion in population screening and the results was randomly checked by direct DNA sequencing. Data were analyzed through association as well as linkage approaches. RESULTS: The Ser20Gly mutation of the IAPP gene was observed in Chinese. It was more prevalent in NIDDM (17 cases, 2.8%) than in non-diabetics (1 cases, 0.5%) (Fisher two-tailed exact P = 0.05). The mutation carrier detected by PCR-RFLP was confirmed to be the A to G point mutation in nucleotide 582 of IAPP gene cDNA encoding the amino acid codon 20. All the mutation carriers detected in population screening were heterozygotes. Analysis of the family members of the 12 NIDDM pedigrees with the IAPP gene Ser20Gly mutation showed that in two families, the mutation was not cosegregated with the affection status. The older was the age of Ser20Gly mutant carrier, the more prevalent was the diabetes in families (P = 0.0001). The highest total lod score of this 12 pedigree was 0.021(theta = 0) in parametric linkage analysis with the model of autosomal dominance with incomplete penetrance. CONCLUSION: The Ser20Gly mutation of IAPP gene was present in Chinese. This mutation does not cause monogenic inheritance diabetes, but may be a pathogenetic factor for the development of NIDDM, the complex genetic disease. PMID- 11038775 TI - [The mechanisms of hyperhomocysteinemia in coronary heart disease]. AB - OBJECTIVE: To identify hyperhomocysteinemia (HHe) as a new and independent risk factor for coronary heart disease (CHD). METHODS: The association of coronary heart disease (CHD) and lipid, homocysteine (HCY) and the factors related to its metabolisms were examined. The mutation of the 677C-->T transition of MTHFR was determined by PCR-based assay. Whole blood and plasma folate (FA) and plasma vitamin B12(B12), as cofactors of those enzymes, were determined by radioimmunologic assay. Plasma HCY was determined by HPLC. RESULTS: Patients with CHD confirmed by coronary angiography had increased plasma HCY concentrations (17.1 +/- 3.6 mumol/L, 7.6 +/- 1.2 mumol/L). In patients with MI, HPT and family history (FH) of CHD, plasma HCY increased significantly. Plasma HCY concentrations had significant non-linear inverse relation with plasma FA and B12 concentrations. Homozygous mutants showed higher plasma HCY concentrations. Patients with CHD had increased serum CHOL and VLDL levels, but plasma HCY concentrations were not correlated with serum lipid levels. CONCLUSION: Hyperhomocysteinemia is an independent risk factor for coronary heart disease. PMID- 11038776 TI - [Ineffective platelet transfusion in patients with hematology malignancy]. AB - OBJECTIVE: To explore the relationship between the effects of platelet transfusion and histocompatibility antigen (HLA). METHODS: The relationship between the corrected count increment (CCI) of platelet with lymphocytotoxicity test (LCT) and human histocompatibity antigen system-I subtype (HLA-I) in 126 hospitalized patients who accepted platelet transfusion. RESULTS: Ineffective platelet transfusion was strongly related to LCT positive and LCT intensity (P < 0.01). The HLA A2 gene frequency was 0.91. The frequency was much higher than that of health human being (0.54). CONCLUSION: Positive LCT was mainly immune factor for ineffective platelet transfusion. CCI and LCT should be the routine test for platelet patients. It may improve the effects of platelet transfusion. PMID- 11038777 TI - [Activation of CD4 T lymphocyte and release of interleukin-5 in airway from patients with atopic asthmatics]. AB - OBJECTIVE: To evaluate the action of CD4T cell and release of interleukin-5 in airway from patients with atopic asthma by the allergen-specific stimulation. METHODS: Twelve atopic asthmatics (AA), 9 atopic non-asthmatics (AN), and 10 normal controls(N) underwent whole-lung inhalation challenge with house dust mite allergen (HDM) extract. The Levels of CD4CD25T lymphocytes, eosinophils(EOS), interleukin-5 (IL-5) and eosinophil cationic protein (ECP) in bronchoalveolar lavage (BAL) and peripheral blood mononuclear cells (PBMC) were investigated with and without HDM challenge. RESULTS: AA differed from AN in having late airway reactions (LAR) after HDM inhalation (P < 0.01), which was correlated with an percentage of BAL eosinophils, CD4CD25T cell IL-5 production and ECP release. CONCLUSION: These findings suggest that the activation of CD4T lymphocytes is correlated with both asthmatic and atopic status. The allergen-specific stimulation is believed to play a major role in CD4 T cell activation in atopic asthma. IL-5 is selective cytokine for EOS activation and modulating local eosinophil recruitment and activation in airway. PMID- 11038778 TI - [The effects of FSH,LH and insulin on steroids production by granulosa cells from polycystic ovaries syndrome]. AB - OBJECTIVES: To investigate steroids production of granulosa cells obtained from normal ovaries and polycystic ovaries from polycystic ovary syndrome (PCOS) patients under FSH, LH and insulin stimulation. METHODS: Granulosa cells of 33 folicles from 18 pairs of normal ovaries and about 100 folicles from 8 pairs of PCOS were cultured. Estrodial (E2) and progesterone (P) accumulation in the medium was determined under different incubated conditions by RIA. RESULTS: Granulosa cells from PCOS produced more E2(P < 0.05-0.01) under basal, FSH and insulin stimulation and more P (P < 0.05-0.01) under FSH, LH and insulin stimulation than controls. Insulin markedly augmented FSH and LH-stimulated steroid accumulation by two groups of granulosa cells. Granulosa cells from PCOS with peripheral insulin resistance also responded to insulin. CONCLUSION: Enzymes for steroids synthesis in granulosa cells from PCOS may be activated in vivo, and hyperinsulinisma and elevated LH levels in PCOS may be contributable. PMID- 11038779 TI - [Correlation between p53 gene mutation and protein expression in 30 cases of human lung cancer]. AB - OBJECTIVE: To investigate the correlation between p53 gene mutation and protein expression and discuss the sensitivities of immunohistochemistry and PCR-SSCP in detecting p53 abnormalities in lung cancer. METHODS: Immunohistochemistry, PCR SSCP, and PCR-sequencing were used. RESULTS: 22 of 30 patients showed p53 mutations by PCR-sequencing. Mutations of 8 patients (36%) located on exon 5, 2(9%) on exon 6, 7 (32%) on exon 7, and 5(23%) on exon 8, respectively. 14 patients (64%) showed missense mutations, and 4(18%) neutral mutations, 3(14%) frameshifts and 1(5%) splice site mutation. In all mutations, G:C-->T:A transversions accounted for 41%(9/22). p53 missense mutations were found in all 10 patients whose immunohistochemistry and PCR-SSCP were both positive. Nine of 10 patients whose PCR-SSCP was positive but immunohistochemistry negative had p53 mutations, including 4 patients with neutral mutations, 3 frameshift mutations, 1 missense mutation, and 1 splice site mutation. In 10 patients whose immunohistochemistry was positive but PCR-SSCP negative, only 3 showed p53 missense mutations. CONCLUSION: Most of p53 mutation sites located on exon 5 and exon 7 in lung cancer of China. Missense mutations and G to T transversions were prevalent. If both PCR-SSCP and immunohistochemistry gave the positive results, p53 missense mutations may be expected. PMID- 11038780 TI - [TGF-beta 1 antisense gene transfer into ito cells and suppressed extracellular matrix production]. AB - OBJECTIVE: To investigate possible role of antisense TGF-beta 1 RNA in the regulation of TGF-beta 1 and ECM production in Ito cells. METHOD: A human transforming growth factor-beta 1(TGF-beta 1) cDNA (1467 bp) was inserted in reverse orientation into the retroviral vector, constructed retroviral vector pLATSN of antisense RNA for TGF-beta 1. A higher-titer, recombinant retroviral vector carried antisense RNA for TGF-beta 1 produced in PA317 packaging cells has been introduced into human Ito cells lines LI90. After selection with G418, resistant colonies were obtained. RESULTS: Stable integration of retrovirus in infectants was shown the presence of antisense RNA was detected by RT-PCR. The expression of TGF-beta 1 protein and the production of extracellular matrix such as FN, Co1A1 were markedly decreased in the antisense TGF-beta 1 transfected cultured cells by ELISA, immunohistochemistry and in situ hybridization. CONCLUSION: Antisense RNA of TGF-beta 1 can be successfully used to inhibit Ito cells activated, endogenous TGF-beta 1 mRNA and extracellular matrix produced, and may provide a basis for the development of anti-fibrosis gene therapy. PMID- 11038781 TI - [The effect of antisense epidermal growth factor receptor RNA on the growth of human malignant glioma cells]. AB - OBJECTIVE: To study the effect of antisense EGFR RNA on the growth of human malignant glioma cells. METHODS: An antisense EGFR expression vector was constructed and introduced into human malignant glioma cells. PCR analysis was used to determine which clones were successfully transfected with antisense EGFR constructs. Western blotting analysis was used to determine EGFR protein level in the transfected cells. The in vitro growth and their ability to grow in soft agar were studied. RESULTS: Western blotting analysis showed six clones stably expressing lower levels of EGFR protein. Antisense EGFR RNA was found to inhibit the proliferation of human malignant glioma cells and their ability to grow in soft agar. There was a negative correlation between the growth inhibiting effect of antisense EGFR and EGFR protein expression. CONCLUSION: These results demonstrate that EGFR play an pivotal role in human malignant glioma cell growth. Antisense EGFR can inhibit the growth of human malignant glioma cells. PMID- 11038782 TI - [Antifertile effect of immunization with anti-idiotypic antibodies to porcine zona pellucida]. AB - OBJECTIVE: To evaluate the antifertile effect of anti-idiotypic antibodies to PZP. METHODS: The New Zealand rabbits were immunized by 17D3 monoclonal antibody against PZP to produce anti-idiotypic antibodies (Ab2) which were then purified by immuno-affinity chromatography. RESULTS: The mice immunized with Ab2 showed decrease of pregnancy rate and a statistically significant reduction in the number of embryos as compared to controls (P < 0.05). Histological examination of ovaries showed that Ab2 interferes only with late follicular development. CONCLUSIONS: Ab2 to 17D3mAb could alleviate ovarian damage and suppress partial fertility. Clearly, further research is required to screen epitope of inner image of Ab2 and to express it by gene engineering. PMID- 11038783 TI - [The effect of immunosuppression on the expression of perforin and granzyme B mRNA in hamster to rat liver transplantation]. AB - OBJECTIVE: To explore the variation on the expression of perforin and granzyme B gene mRNA to judge the effect of immunosuppression in acute rejection of liver transplantation. METHODS: The expression of perforin and granzyme B gene mRNA was examined by RT-PCR in hamster to rat liver grafts under the immunosuppression of cyclosporine and splenectomy. Histological findings were studied comparatively. RESULTS: Cyclosporine and splenectomy could obviously suppress the rejection of liver grafts. The survival time of animals was significantly prolonged (37.1 days). The architecture of hepatic lobule was preserved. There was slight round cell infiltration in the portal tracts and no expression of mRNA of perforin and granzyme B genes could been seen in three weeks after transplantation. CONCLUSION: Perforin and granzyme B genes are of value in judging the effect of immunosuppression. PMID- 11038785 TI - [Relation between autopsy and increasing the level of medicine]. PMID- 11038784 TI - [Autopsy]. PMID- 11038786 TI - [Conference on the importance of autopsy under modern medical conditions]. PMID- 11038787 TI - [Total body irradiation etoposide followed by autologous hematopoietic stem cell transplantion for non-Hodgkin's lymphoma]. AB - OBJECTIVE: To evaluate the therapeutic effectiveness and safety of total body irradiation(TBI) and Etoposide(Vp-16) as a preparative regimen for autologous hematopoietic stem cell transplantation(AHSCT) with non-Hodgkin's lymphoma(NHL). METHODS: Twenty-four patients with intermediate and high grade NHL underwent AHSCT. They achieved complete remission (CR) or partial remission (PR) after induction chemoradiotherapy. Twenty-three patients had first CR or PR, and one third CR. Ten patients underwent autologous bone marrow transplantation (ABMT) and 14 autologous peripheral blood stem cell transplantation (APBSCT). The preparative regimen was TBI 800(700-850) cGy/Vp-16 757(323-1140) mg/m2. RESULTS: At a median follow-up of 22.5(2-92) months, the one-year disease free survival(DFS) was 86.7% (13/15), 3-year, 5-year and 7-year DFS were 80.0% (12/15) in patients with CR before AHSCT. The DFS was 66.7% (4/6) in patients with PR before AHSCT. The patients who had relapsed before AHSCT(3 cases) did not reach DFS. The hematopoietic function recovery was rapid in APBSCT than ABMT. CONCLUSION: The clinical results of AHSCT for intermediate and high grade NHL who achieved CR or PR after induction therapy are satisfactory. The TBI/Etoposide is an effective and safe preparative regimen for AHSCT in NHL patients. PMID- 11038788 TI - [Insulin receptor substrate-1 and glucose transporter gene polymorphisms in noninsulin-dependent diabetes mellitus]. AB - OBJECTIVE: To evaluate the role of insulin receptor substrate-1 (IRS-1) and glucose transporter-1 (GLUT1) gene polymorphisms in Chinese patients with noninsulin-dependent diabetes mellitus (NIDDM) and diabetic neophropathy (DN). METHODS: Aminoacid polymorphism in codon 972 of IRS-1 gene and the polymorphic Xba I site of GLUT1 gene were analyzed by PCR-RFLP in 131 patients with NIDDM and 124 normal subjects. DN was defined as persistent albuminuria and/or impaired renal function, without known cause of renal diseases other than diabetes. Insulin sensitivity index (ISI) and body mass index (BMI) were also calculated. RESULTS: The distribution of IRS-1 gene polymorphism showed no difference between patients with NIDDM and normal controls. The frequencies of Xba I (+/-) genotype (59% vs. 33%, P < 0.01) and Xba I (-) allele (37% vs. 21%, P < 0.01) were significantly higher in NIDDM patients than in normal subjects. To further explore the linkage of GLUT1 gene polymorphism with DN, we examined the GLUT1 genotype of NIDDM patients with or without renal damage. The frequency of Xba I (+/-) genotype (75% vs. 44%, P < 0.01) and Xba I (-) allele (44% vs. 29%, P < 0.05) was significantly higher in NIDDM patients with diabetic nephropathy than either those without nephropathy or normal subjects. However, there were no significant differences of GLUT1 genotype and allele frequency in NIDDM patients without nephropathy and normal controls. The presence of Xba I (-) allele appeared to have a strong association with the development of diabetic nephropathy. The odds ratio was 1.915, and the 95% confidence interval was 1.044 3.514. The association of Xba I (-) allele of GLUT1 gene with NIDDM partly reflected their close association with DN. Although there was no correlation between the gene polymorphism of GLUT1 and BMI, patients carrying the Xba I (-) allele showed a lower ISI. CONCLUSION: No association was found between the gene polymorphism of IRS-1 and NIDDM. THe Xba I (-) allele of GLUT1 gene might be taken as a genetic marker of NIDDM with diabetic nephropathy and this genetic susceptibility appears to be associated with the insulin resistance in patients with NIDDM. PMID- 11038789 TI - [Evaluation of thoracic aortic anatomy and function with transesophageal echocardiography]. AB - OBJECTIVE: To evaluate the effects of age, gender, and hypertention on thoracic aortic anatomy and function in Chinese. METHODS: Thoracic aortic diastolic and systolic diameter (Dd and Ds), intimal-medial thickness (IMT), Peterson's and Young's elastic modulus (Ep and Es), and stiffness index (beta) were measured by transesophageal echocardiography in 134 normal subjects and in 42 patients with hypertension. RESULTS: Age was positively correlated with IMT (r = 0.68, P < 0.01), Dd(r = 0.65, P < 0.01), Ds(r = 0.58, P < 0.01), Ep(r = 0.82, P < 0.01), Es(r = 0.71, P < 0.01), beta(r = 0.67, P < 0.01). Men had a greater diameter and IMT than women. In comparison with the control group, the patients with hypertension had significantly increased Dd (20.73 +/- 2.80 mm vs 18.20 +/- 3.03 mm, P < 0.01), Ds (22.17 +/- 2.90 mm vs 19.81 +/- 2.93 mm, P < 0.01), IMT(1.39 +/ 0.66 mm vs 1.17 +/- 0.50 mm, P < 0.05), Ep(1.47 +/- 0.84 vs 0.81 +/- 0.66, P < 0.01), Es(11.91 +/- 8.04 vs 6.49 +/- 3.93, P < 0.01), and beta(3.24 +/- 0.43 vs 2.91 +/- 0.52, P < 0.01). CONCLUSIONS: Aging and hypertension significantly increase aortic diameter, thickness and stiffness. Men have higher aortic diameter and wall thickness than women but similar stiffness. PMID- 11038790 TI - [Diagnostic evaluation of magnetic resonance urography in urinary tract dilatation and obstruction]. AB - OBJECTIVE: To evaluate the diagnostic value of magnetic resonance urography (MRU). METHODS: Two hundred and one patients were examined, 92 patients with urinary tract dilatation using a modified, heavily T2-weighted fast spin-echo pulse sequence and fat-suppression pulse aid. Postprocedure processing was performed with a maximum-intensity-projection (MIP) algorithm. RESULTS: In the 92 patients with urinary tract dilatation, 86 had obstructive urinary tract dilatation 6 non-obstructive urinary tract dilatation. In the 86 patients, 14 had ureteral carcinoma, 19 calculus, and 13 benign stricture, 23 congenital ureteral stricture/or with anomalies, 7 ureteral obstructive caused by extrinsic pelvis disease, and 10 bladder carcinoma involved ureter. In the 6 patients, 5 had nervous bladder. MR urography provided the high-resolution image of the urinary tract and determined the presence of obstruction. CONCLUSION: MR urography is a reliable noninvasive method for depicting the urinary tract. PMID- 11038791 TI - [Brain evoked potentials in patients with Alzheimer's disease and schizophrenia: a comparative study]. AB - OBJECTIVE: To investigate the differences of visual evoked potentials(VEP) and auditory evoked potentials(AEP) between patients with Alzheimer's disease and schizophrenics. METHODS: Both VEP and AEP were elicited from 39 patients with Alzheimer's disease, 34 aged schizophrenics and 40 normally age-matched subjects. RESULTS: The variability of waveforms of both VEP and AEP became increased within the AD group. The AD patients presented significantly delayed latencies (N2 and P3 of VEP; P1, N1, P2, N2 and P3 of AEP) as compared to the normal controls. The P2 amplitude of VEP of AD patients was 32% lower than that of schizophrenics and 57% lower than that of the controls. The P2 amplitude of AEP of AD patients was 36% lower than that of schizophrenics and 70% lower than that of the controls. Compared with the normal controls, the schizophrenics showed reduced VEP-P2 latency, prolonged N2 and P3 latencies of both VEP and AEP. The P2 amplitudes of VEP and AEP of schizophrenics were decreased by 16% and 20% respectively as compared to the normal controls. CONCLUSION: The variations of VEP and AEP are a referential tool for diagnosing Alzheimer's disease and schizophrenia. PMID- 11038792 TI - [Gastro-intestinal reconstruction by substitution of "O" shaped jejunum after total gastrectomy]. AB - OBJECTIVE: To discuss a satisfying method for alimentary continuity reconstruction after total gastrectomy. METHOD: An "O" shaped jejunum pouch or a new "stomach" was constructed in between esophagus and duodenum after total gastrectomy. RESULTS: Thirty-nine cases of gastric malignant tumor were treated by this method. The 3 and 5 years disease free survival rate were 67.7% and 55.6%. All the patients recovered their normal meal habit (3 meals a day) within 6 months postoperatively. Compared with the preoperative and the postoperative (one year after operation), body weight is equal level, increasing and decreasing in 71.1%, 15.8% and 13.1% of the cases respectively. There were no posterior sternal pain and food or digestive fluid reflux, no dumping syndrome in these patients. The volume enlargement of the "new stomach" was observed obviously, especially in "stomach fundus". CONCLUSION: This operation is a satisfying method for alimentary canal reconstruction after total gastrectomy and it is worthy to be recommended in gastric surgery. PMID- 11038793 TI - [The effects of nimodipine on cerebrovascular function in acute cerebral infarction]. AB - OBJECTIVES: To use cerebrovascular hemodynamics indexes (CVDI) to evaluate the effects of nimodipine on cerebrovascular function in acute cerebral infarction (ACI), and to investigate the changes of blood pressure (BP), heart rate (HR), and electrocardiography (ECG). METHODS: 30 patients with ACI in internal carotid artery system within 72 h from the oneset of symptoms were given nimodipine (10 mg, 0.25-0.5 microgram/kg/min) by intravenous drip (i.v.) every day for 14 days. CVDI, BP, HR and ECG were examined at various times during treatment. RESULTS: Nimodipine improved cerebrovascular function significantly. Cerebral blood velocity and flow increased, cerebrovascular resistance and criticle pressure decreased, and cerebrovascular elasticity and autoregulation improved. BP dropped significantly at the third day, and mainteined stable afterward. HR dropped remarkably at the fourteen day (P = 0.011). It was reasonable that the treatment of nimodipine (i.v.) lasted 10 days or so. CONCLUSIONS: Nimodipine improves cerebrovascular function significantly, but monitoring of BP and ECG is necessary during its application. PMID- 11038794 TI - [Changes of bone and mineral metabolism in patients with hyperthyroidism before and after treatment]. AB - OBJECTIVE: To investigate the changes of bone and mineral metabolism in patients with hyperthyroidism before and after treatment. METHODS: Enzyme linked immunosorbent assay was used to measure bone-specific alkaline phosphatase (BAP) and deoxypyridinoline (DPD); radioimmunoassay to to measure serum calcitonin (CT), parathyroid hormone (PTH-M), bone GLA protein (BGP) and other markers related to bone metabolism; dual energy X-ray absorptiometry to measure bone mineral density (BMD) of spine(L2-4) and femur(Neck, Troch, Ward's) in 45 patients with hyperthyroidism before and after treatment and 58 healthy volunteers. RESULTS: The urinary DPD level was elevated by 527% in patients with hyperthyroidism before treatment. Compared to controls the serum ALP, BAP, BGP elevated by 62%, 146%, 87% (P < 0.001), BMD decreased to various extent, and women L2-4, Ward's were marked (P < 0.05). The results after treatment and before treatment showed that urinary DPD decreased by 79%; serum ALP, BAP, BGP decreased by 19.5%, 24.7%, 27.5% respectively (P < 0.001, 0.05, 0.05, > 0.05). All sites BMD increased, and women Troch sites were marked (P < 0.05). The correlation analysis showed that the urinary DPD was positively correlated with serum BGP(r = 0.349 P < 0.05). FT4 was positively correlated with BAP, ALP and DPD respectively) (r = 0.353, P < 0.05, r = 0.294 P = 0.05, r = 0.426 P < 0.01). CONCLUSION: Hyperthyroidism bone disease is caused by excessive serum thyroid hormone that speeds up bone turnover and marked bone absorption compared to bone formation. PMID- 11038795 TI - [Cardiovascular protective effects and NO-mediated cerebrovasorelaxant effects of extract of ginkgo biloba leaves]. AB - OBJECTIVE: To examine the anti-free radical related cardiovascular protective effects and NO-mediated cerebrovasorelaxant effects of extract of the leaves of Ginkgo biloba(EGb) in isolated preparations. METHODS: Experiments on the anti free radical related cardiovascular protective effects were performed in rabbit Langendorff heart and isolated aortic rings damaged by diphenyl-picryl hydyazyl(DPPH). Protective effects of EGb were determined by comparing the results of EGb pretreated group with the DPPH-injured controls. Cerebrovasorelaxant effects of EGb were examined in ring preparations of isolated porcine basilar artery in vitro using tissue bath techniques. RESULTS: EGb protected the isolated rabbit heart from the DPPH-injury to the cardiac contractility and the aortic endothelium from DPPH-attenuated ACh-induced relaxation. EGb concentrationde dependently relaxed the basilar artery and this effect was more significant in endothelium intact rings than those endothelium denuded rings. EGb enhanced the TNS-induced relaxation in basilar artery and this effect was abolished by pretreatment of N-nitro-L-arginine or tetrodotoxin. CONCLUSIONS: EGb exerts cardiovascular protection against DPPH-impaired cardiac contraction in rabbit isolated heart and endothelium-dependent relaxation in the aortic ring of rabbit in vitro. EGb relaxes porcine basilar artery and enhances the TNS-induced relaxation via a NO pathway. PMID- 11038796 TI - [Ischemia and reperfusion reduce the mRNA expression of endogenous basic fibroblast growth factor (bFGF) in rat skeletal muscles]. AB - OBJECTIVE: To explore the expression of bFGF gene in normal and wounded rat skeletal muscles. METHODS: In situ hybridization and quantitative PCR methods were used to evaluate the location of basic fibroblast growth factor (bFGF) mRNA and the amount of bFGF mRNA expression in normal, ischemic or ischemic and reperfused rat skeletal muscles. RESULTS: According to the intensity of the stain in situ hybridization, four main classes of fibers could be identified: strong, moderate, weak and negative stained fibers. bFGF mRNA was found in the cytoplasm in both normal and wounded muscles; however, the signal intensity was much stronger in normal muscles. In the normal muscles, bFGF mRNA was distributed irregularly and sparsely, and 82% of fibers had positive bFGF mRNA staining, and only 52% and 22% of fibers in ischemic or ischemic and reperfused muscles were positive for bFGF mRNA staining in situ hybridization. The results in quantitative PCR study supported the concept of the reduced bFGF mRNA expression in the wounded muscles. CONCLUSIONS: The loss of stored bFGF in wounded skeletal muscles as we showed previously is also caused by its increased damage and reduced secretion. PMID- 11038797 TI - [Preventional intervention of myocardial interstitial fibrosis in murine myocardium with acute myocarditis]. AB - OBJECTIVES: To explore that whether intervening the fibrosis of heart interstitium in acute viral myocarditis is practical or not, and offer the experimental basis for clinic to choose the optimum period to intervene myocardial fibrosis. METHODS: Animals were divided into three groups: myocarditis, myocarditis intervened by Losartan, and normal control. The death rate of every group was compared. HE staining and picrosirius red staining and circularly polarized light were used to investigate myocardial collagen expression. Cardiac output was measured by impedance differentiation, and cardiac index was computed to evaluate cardiac function. RESULTS: The death rate of the group with Losartan was 75.0%, and that of the myocarditis group was 41.7%. The cardiac index of the group with the drug was 0.014 +/- 0.001 ml.min-1.cm2, the myocarditis group 0.019 +/- 0.004 ml.min-1.cm2, and the normal control 0.024 +/- 0.002 ml.min-1.cm2. The scanning area of collagen in the group with the drug was 2.06 +/- 0.77 mm, the myocarditis group 4.72 +/- 2.22 mm and the normal control 3.74 +/- 1.50 mm. CONCLUSIONS: Collagen has increased in acute viral myocarditis, but its main role in this time is to repair the necrotic myocardium, and Losartan may block this process and make lesions develop. Therefore, the acute stage is not an practical period for intervening myocardial fibrosis in viral myocarditis. PMID- 11038798 TI - [Expression of endothelin-1 and nitric oxide synthase mRNA in gastric mucosa of rats with cirrhosis and portal hypertensive gastropathy after disconnective operations]. AB - OBJECTIVE: To investigate expression of endothelin-1 (ET-1) and nitric oxide synthase(NOS) gene in gastric mucosa in the rats with cirrhosis and portal hypertensive gastropathy (PHG) after disconnective operation. METHODS: Wistar rats were divided into three groups: normal (n = 12), cirrhosis and PHG (n = 12), and post-disconnective (n = 12). The rat models were induced by intraperitonal injection of thiocetamide for 16 weeks. The third group rats were reared for 3 months after the operations. ET-1 and NOS mRNA from the gastric mucosa of the three groups were measured quantitatively by Dot blot technique. RESULTS: ET-1 mRNA decreased in the PHG rats as compared to the nomal rats (P < 0.01). Further decrease was found in the post-disconnective rats (P < 0.01). NOS mRNA increased slightly in the PHG rats (P > 0.05) and increased obviously in the post disconnective rats (P < 0.01). CONCLUSIONS: After the cirrhosis and PHG rats were subjected to disconnective operations, ET-1 was insufficient and NOS was too much in the gastric mucosa. These resulted in the gastric mucosal vessel dilatation. The imbalance of ET/NO proportion could partly attribute to rebleeding after disconnection. PMID- 11038799 TI - [The inhibitory effect of melatonin on morphine withdrawal syndromes and serum monoamines in morphine dependent mice]. AB - OBJECTIVE: To observe the inhibitory effect of melatonin on morphine withdrawal syndromes and serum monoamines in morphine dependent mice. METHODS: A physical dependent model in mice was established by subcutaneous injection of morphine. The intensity of withdrawal syndromes was evaluated according to the jumping latency and jumping times. The concentration of serum monoamines was detected with HPLC-ECD. RESULTS: The physical withdrawal syndromes in morphine dependent mice were inhibited partly by four different doses (25, 50, 100, 200 mg/kg) of melatonin and showed a significant dose-dependent manner. The increased concentration of serum norepinephrine and dopamine in morphine-dependent mice could be reduced by large dose (100 mg/kg) of melatonin. CONCLUSION: The jumping withdrawal syndromes and serum monoamiues in morphine-dependent mice could be inhibited partly by melatonin. PMID- 11038800 TI - [Relation between insulin-like growth factor II gene and liver cancer]. PMID- 11038801 TI - [Research on the relation between apoptosis imbalance and diseases]. PMID- 11038802 TI - [Brain cell apoptosis after cerebral hypoxia-ischemia in neonatal rat]. AB - OBJECTIVE: To investigate action of apoptosis in mechanisms of neonatal hypoxia ischemia (HIE). METHODS: Using HE staining, electronmicroscope, and terminal deoxynuleotidyl transferase-mediated dUTP nick end labeling (TUNEL) technique, we observed the histological features, time course and density of apoptotic cells and compared the ipsilateral hemisphere after permanent ischemia 1, 4, 18, 24, 40, 72 h, 7 days after 3 h hypoxia with that of sham-group. RESULTS: Neurons in the cortex, hippocampus, thalamus exhibited cells shrink, chromatin condensation, apoptosis bodys under microscope, as demonstrated by in situ labeling of DNA breaks. The peak for apoptosis was shown at 18 h in the cortex. CONCLUSION: In the HIE, apoptosis appears earlier than necrosis, and both of the different cell death forms may exist induced dose relativity. PMID- 11038803 TI - [The deletion of AT2 receptor gene antagonizes angiotensin II-induced apoptosis in fibroblasts]. AB - OBJECTIVE: To study the expression of angiotensin II (Ang II) type 2 receptor (AT2) and its effect on the Ang II-induced apoptosis in fibroblasts. METHODS: Skin fibroblasts were cultured from the embryos of the AT2 receptor gene deleted (AT2-/-) and wild-type (AT+/+) mice at 16 days of gestation. After Ang II being stimulated, the expressions of AT1 and AT2 receptor genes were examined by RT-PCR and the apoptotic changes were identified by genomic DNA electrophoresis, TUNEL staining and flow cytometry analysis of DNA contents. And they were compared in these two types of cultured fibroblasts. RESULTS: Treated with Ang II for 48 h at the concentrations of 10(-8), 10(-7), 10(-6) and 10(-5) mol/L respectively, the expressions of AT1 and AT2 receptor genes were enhanced in a dose-dependent manner. Induced by Ang II for 72 h at the concentrations of 10(-6) and 10(-5) mol/L, genomic DNA fragmentation, one of convinced markers of cell apoptosis was found in the AT2+/+ fibroblasts, but not in the AT2-/- fibroblasts. The percentages of Ang-induced apoptosis of the AT2+/+ fibroblasts were (12.3 +/- 2.7)% (10(-6) mol/L) and (21.7 +/- 6.7)% (10(-5) mol/L) respectively. CONCLUSION: The Ang II-induced apoptosis of fibroblasts was devoid after deletion of AT2 receptor gene. PMID- 11038804 TI - [Apoptosis versus proliferation activities and relative mechanism in chronic obstructive pulmonary disease]. AB - OBJECTIVE: To study the roles of apoptosis and proliferation and relative genes expression in chronic obstructive pulmonary disease (COPD) and pulmonary hypertension. METHODS: Immunohistochemical technique was used for the detection of cell proliferation and expression of relative genes. In situ end labeling technique was used for the detection of nucleosomal DNA fragmentation of apoptotic cells, and Northern blot for the detection of expression of c-myc, bcl 2 and p53 genes in the lungs with COPD. RESULTS: Both proliferative cells and apoptotic cells were found in the lungs with COPD or without COPD. In lung tissue with COPD. The proliferation index was increased significantly, whereas the apoptosis index was decreased significantly. Compared with controls, the ratio of proliferation to apoptosis in lungs with COPD was increased by 4 folds, and the expression of c-myc or bcl-2 mRNA was significantly increased by 2.5-3 folds in lungs tissue with COPD. The expression of c-myc and bcl-2 protein was also increased significantly in lungs with COPD, the two antigen were predominantly localized in small pulmonary vessels. The expression of p53 mRNA was significantly decreased by 3 folds in lung tissue with COPD. CONCLUSION: The abnormality of apoptosis versus proliferation activities induced by c-myc, bcl-2 and p53 genes may contribute to pulmonary vascular structural remodeling in COPD. PMID- 11038805 TI - [Amyloid protein precursor gene and neuron apoptosis of Alzheimer's disease]. PMID- 11038806 TI - [Hyperinsulinemia and insulin resistance in nondiabetic first-degree relatives of patients in familial NIDDM pedigrees]. AB - OBJECTIVE: To study plasma insulin level and insulin sensitivity of nondiabetic first-degree relatives of patients in familial non-insulin-dependent diabetes mellitus (NIDDM) pedigrees. METHODS: Standard oral glucose tolerance test(OGTT) was conducted in 55 healthy controls and 97 first-degree relatives of NIDDM probands in familial NIDDM pedigrees. Plasma glucose and plasma insulin were determined at all phases after oral glucose load. Moreover, we calculated the glucose/insulin area ratio under the curve of OGTT (SGI) and the insulin sensitivity index (ISI) in all subjects to estimate their insulin sensitivity. RESULTS: In nondiabetic first-degree relatives, SGI and ISI were significantly decreased (P < 0.01), and plasma insulin was increased at 0, 30, 60, 120 minutes after administration of oral glucose load (P < 0.05 and P < 0.01), as compared with the controls, in matched age, body mass index (BMI) and proportion of sex. In both first-degree relatives and the insulin controls, SGI was positively correlated with ISI (r = 0.39, 0.41, respectively, P < 0.01) and both SGI and ISI were inversely correlated with BMI (r = -0.38, -0.39; -0.32, -0.25, respectively, P < 0.01). Moreover, FINS in the two groups was positively correlated with BMI (r = 0.30, 0.34, respectively, P < 0.05). CONCLUSION: In nondiabetic first-degree relatives of patients in familial NIDDM pedigrees, plasma insulin level was increased and insulin sensitivity was decreased as compared with the healthy controls. It suggests that they suffered from hyperinsulinemia and insulin resistance. PMID- 11038807 TI - [Olfactory event-related potentials to isoamyl acetate in congenital anosmia]. AB - OBJECTIVE: To investigate the olfactory origin of the event-related potential (OEP) by evaluating OEP and standardized psychophysical measures in patients with congenital anosmia. METHODS: Olfactory function was evaluated by OEP and standardized psychophysical measures including smell identification test and odor detection threshold tests for 3 chemosensory stimulants: phenylethyl alcohol (PEA), isoamyl acetate (IAA), and chloracetyl phenone (CAP) in 9 subjects with isolated congenital anosmia and 9 age- and gender-matched normosmic controls. RESULTS: There was a significant difference in the smell identification test score (P < 0.001) and odor detection thresholds for PEA and IAA (P < 0.001) between the anosmic and normosmic subjects. Detection thresholds for CAP, a trigeminal stimulant, did not differ between the 2 groups. In the control subjects, OEP to IAA was characterized by 4 reproducible components (P1, N1, P2 and N2). In the subjects with congenital anosmia, no reproducible evoked potential components were identified in response to IAA. No reproducible evoked potential components were seen in response to the air control stimulus in either the anosmic or normosmic groups. CONCLUSION: These data suggest that OEP provide a specific and objective measure of olfactory function. PMID- 11038808 TI - [HLA-DRB alleles polymorphism in susceptibility to asthma in Beijing Chinese]. AB - OBJECTIVES: To investigate whether susceptibility or resistance to asthma associated with HLA-DRB alleles and analyze the relationship between HLA-DRB genes and clinical phenotype of asthma (TIgE, sIgE, BHR). METHODS: Using PCR SSP(sequence-specific primer polymerase chain reaction), we tested the frequency distribution of HLA-DRB alleles in 50 asthmatic patients and 80 healthy volunteers from Beijing China. All patients had their serum TIgE, IgE antibody specific to house dust mite measured by RAST, bronchial responsiveness assessed by methacholine bronch-provocation (if FEV1% > or = 70%), and broncho-dilation measurement by inhaling salbutamol. RESULTS: There was significantly increased gene frequency of alleles DR6(13), DR52 in asthmatics compared with normal controls (17% vs 4.3%, P < 0.01; 50% vs 17.5%, P < 0.01), and RR was 7.55 and 4.7 respectively. The frequency of DR2(15), DR51 was lower in asthmatics than in controls (7% vs 18% P < 0.01; 2% vs 33.8% P < 0.01). The percentage of HLA haplotype DR6(13)-DR52 was higher in asthmatics than in healthy volunteers (20% vs 4%, P < 0.01, RR 6.4). 70% of individuals sharing DR6(13) gene and 56% of subjects carrying DR52 gene had elevated serum d1 sIgE antibody (> or = +4). There was no relationship between HLA-DRB alleles and total IgE, BHR. CONCLUSIONS: Alleles DR6(13), DR52 are significantly implicated in their susceptibility to asthma, at least they may be closely associated with this disorder. Conversely DR2(15), DR51 alleles might confer protection against asthma. Positive associations between DR6(13), DR52 and IgE response to d1 allergen are noted. HLA-DRB genes are particularly involved in regulating human atopic immune response. PMID- 11038809 TI - [TGF beta subtypes in hepatocellular carcinoma]. AB - OBJECTIVE: To detect the expression of three TGF beta isoforms and its clinicopathologic significance in hepatocellular carcinoma(HCC). METHODS: Surgical specimens of 52 patients with small HCC resection during 1992-1995 were used to determine the immunoreactivity of TGF beta protein by ABC immunohistochemistry. These specimens were divided into recurrence group(G1) and recurrence group(G2) according to whether HCC recurred two years after resection. The relations with recurrence of HCC were analyzed. RESULTS: The positive rates in G1 and G2 were 85% (17/20) versus 47% (15/32) for TGF beta 1, 90% (18/20) vs. 78% (25/32) for TGF beta 2, and 45% (9/20) vs. 63% (20/32) for TGF beta 3. Significant difference was found in TGF beta 1 and TGF beta 2, but not in TGF beta 3. CONCLUSION: The high immunoreactivity to TGF beta 1 or TGF beta 2 is positively related to the recurrence of HCC. PMID- 11038810 TI - [Detection of HGV NS5 antigen in liver tissue of patients with chronic liver disease]. AB - OBJECTIVE: To investigate the existence of hepatitis G virus (HGV) in liver tissue. METHODS: HGV NS5 antigen was detected by immunohistochemical method in paraffin-embeded liver tissue of autopsy patients with chronic liver disease. RESULTS: Among 110 samples, 32.7% (36/110) had been detected out HGAg in their liver. When serologica marker was used, the detection rate was 21% (4/19) in HNA E, 36% (25/69) in HBV and 32% (7/22) in HCV infectious group, respectively. HGAg expression in hepatocytes was also found pathologically in 22% of 45 patients with active cirrhosis, 43% of 47 patients with hepatocellular carcinoma, and 33% of 18 patients with chronic fulminent hepatitis. The staining signal of HGV NS5 antigen was mainly located in the cytoplasm of liver or neoplasm cells, and the positive cells were distributed diffusely in pseudolobule or liver tissues. CONCLUSION: The infection of viral G is often seen in liver tissue of patients with chronic liver disease. PMID- 11038811 TI - [Comparison of different surgical procedures for urinary stress incontinence]. AB - OBJECTIVE: Comparison of different surgical procedures for urinary stress incontinence (USI). METHODS: 95 patients with and/or accompanying USI were treated operatively in a prospectively randomized manner. RESULTS: The cure rate of colposuspension group 3 months after operation was 93% and was higher than 69% after Kelly operation (P < 0.05). But the cure rate 12 months after operation decreased to 74% and 58% respectively (P > 0.05). The percentage of abnormal sexual life was lower in the colposuspension group (4%) than that of those having Kelly operation (7%). The average days and times of intubation in the colposuspension group were more than in Kelly operation group (P < 0.05). CONCLUSION: The cure rate of colposuspension 3 months was better than that of the Kelly operation. The cure rate at 1 year was similar in the two procedures. Transient urinary retention occurred more after in the colposuspension group. PMID- 11038812 TI - [Arterial plasma nitric oxide and lactate level in hemorrhagic shock patients]. AB - OBJECTIVE: To investigate the relationship between arterial plasma nitric oxide(NO), lactate (LA) concentration and prognosis of hemorrhagic shock patients. METHODS: The blood levels of a NO and LA were measured with fluoropholometry and colorimetry in 48 hemorrhagic shock patients, and other 30 patients for selective surgery served as controls. RESULTS: NO level was significantly lower and LA level was significantly higher in the hemorrhagic shock group than that of the control group respectively. NO level had a negative correlation with LA level and trauma index. NO level in the patients complicated by sepsis was still lower than that of the control group. CONCLUSION: Decrease of NO level may be a main cause of disturbance of microcirculation and increase of LA. The lower the NO level, the higher the LA level, the worse the prognosis. So NO and LA level play an important role in forecasting progress of hemorrhagic shock patients. PMID- 11038813 TI - [Application of bone graft in revision total hip arthroplasty]. AB - OBJECTIVE: To compare the differences of bony union in autograft, allograft and their mixture-graft. METHODS: The postoperative X-ray films of revision total hip arthroplasties with acetabular bonegraft were observed in 41 patients. They were women, aged at operation from 44 to 86 years (average 65 years). Follow-up ranged from 12 to 72 months (average 29 months). At the operation, autogenous bone was taken from the iliac-crest and homologous bone was from banked bone i.e. femeral head taken previously. Of these, 18 were autografts, 8 allografts, and 15 mixture grafts. Bone incorporation was observed postoperatively on 6-week, 3-month, 6 month, 12-month and over 12-month films. RESULTS: At 6 weeks, graft absorption appeared in 31% patients (13/41). At 3 months, bridging trabeculation across the graft-host interface was present in 88% patients, (36/41). At 6 months, graft remodelling began in 39% patients (16/41). At 12 months, graft remodeling was considered in 37 patients, and radiolucent prothesis in 4 (infection in 3 patients and graft absorption). CONCLUSION: No difference is observed in the time and the condition of bone union between autograft and allograft, autograft or allograft and their mixture graft. PMID- 11038814 TI - [Thallium poisoning a clinical analysis of 5 cases]. AB - OBJECTIVE: To sumarize the clinical features, diagnosis and treatment of thallium poisoning by analysis of clinical cases. METHODS: Five cases of thallium poisoning were reported. The clinical manifestations, quantitative-analysed thallium level, methods and effect of treatment, and prognosis were analysed. RESULTS: Three of the 5 cases were acute, and the other two chronic. Four of them had a history of poison contact. The clinical features included alopicia (4 cases), transverse white stripes in the nails (Mee's stripes, 2) polyneuropathy, cranial nerve and central nervous system impairments (5), and abdominal pain and other digestion disturbances (3). All of the 5 cases were treated with antidotes and other methods to increase thallium excretion. One case died of lung infection 3 days after diagnosis. The serum and urine thallium levels of the 4 survivors decreased to normal after treatment (3 recovered, one had neurologic sequelae). CONCLUSION: It is not difficult to diagnose thallium poisoning by definite poison contact history, classical manifestation, and serum or urine thallium quantitative analysis. Attention should be paid to those patients with indefinite exposure to thallium. For those suspected cases, it is necessary to measure the thallium level in blood and urine in order to verify the diagnosis. PMID- 11038816 TI - [Morphologic changes of hippocampal neurons irradiated with gamma knife]. AB - OBJECTIVE: To study the early radiobiological response of Gamma knife to the cultured hippocampal neurons in vitro. METHODS: The light and electron microscopy were used to see the Gamma knife irradiated neurons of rat. RESULTS: Degenerated neurons showed swelling and refrangibility weakening or disappearance of soma, pellet-like changes of cytoplasm, thickening and breaking of processes under phase-contrast inverted microscopy. Lesions with swelling of soma and fold vanish on cell surface, enlargement of volume and rapture of membrane, the sparse of network among the cells, thickening and breaking of processes were observated under scanning electron microscopy. The ultrastructure of the insulted neurons showed irregular nuclei and concavity of nucleus membrane, a lot of swelling or condensation of mitochondria, separation of ribosome and neurofilament, and vacuolated changes of cytoplasm. CONCLUSIONS: The effect on Gamma knife irradiated neurons is similar to that of normal brain tissues. There is a dose response relationship at early stage. The study indicates that low-dose irradiation can cause damage to cells. PMID- 11038815 TI - [Somastostatin and growth hormone in preventing liver damage due to acute necrotizing pancreatitis]. AB - OBJECTIVE: To study the role of somastostatin and growth hormone in preventing liver damage due to ANP. METHODS: The roles of inflammatory mediators (Endotoxin, Amylase, TNF alpha) were investigated in ANP model by retrograde injection of 3.5% sodium taurocholate 2.5 ml/kg into the pancreatic duct, and TNF alpha mRNA in the liver after ANP was observed by reverse transcriptase-polymerase chain reaction. Besides, the effects of somastostain and growth hormone were also observed. RESULTS: ANP caused remarkable elevation of those inflammatory mediators, being positively correlated with the development of pancreas and liver damage. Somastostain and growth hormone inhibited the inflammatory mediates and TNF alpha mRNA overexpression and reduced the damage to the pancreas and liver. CONCLUSIONS: TNF alpha plays an important role in ANP progression, and somastostatin and growth hormone may prevent the development and progression of liver damage due to ANP. PMID- 11038817 TI - [Interleukin-12 gene transfection into murine B16 melanoma cells suppresses tumorigenicity and decreases metastatic potential]. AB - OBJECTIVE: To study the tumorigenicity and metastasis of poorly immunogenic murine B16 melanoma cells transfected with murine interleukin-12 (IL-12) gene. METHODS: Two recombinant vectors containing the full length cDNA of p40 or p35 subunit of IL-12 were constructed. They were cotransfected into B16 melanoma cells by LipofectAMIN method. The expressions of p40 and p35 mRNAs were analyzed by means of reverse transcription polymerase chain reaction (RT-PCR). The secretion of bioactive IL-12 was detected by bioassay (measuring the proliferative response of PMA-activated murine splenocytes). To determine the effects of IL-12 secreted by genetically engineered B16 cells (B16T-IL12) on tumorigenesis and metastasis, the cells were inoculated s.c. or i.v. into C57BL/6 mice. RESULTS: B16T-IL12 cells expressed both p40 and p35 mRNAs, also produced bioactive IL-12. Only 70% of the mice injected with B16T-IL12 cells developed palpable tumors. The emergence of palpable tumors in the mice inoculated with B16T-IL12 cells was significantly delayed (P < 0.01), the growth rate of the tumors was obviously decreased (P < 0.01), and the survival time of tumor-bearing mice was prolonged substantially (P < 0.01), compared with the control B16 or B16Tneo cells. The incidence of experimental pulmonary metastasis of B16T-IL12 cells was inhibited, and the number of pulmonary metastases was reduced markedly, in contrast to the i.v. inoculation of B16 cells (P < 0.01). CONCLUSION: Transfection of IL-12 gene into B16 cells results in the inhibition of tumorigenesis and in the suppression of experimental pulmonary metastasis. PMID- 11038818 TI - [The protective effects of baicalin on pertussis bacilli-induced brain edema in rats]. AB - OBJECTIVE: To evaluate the inhibitory effect of baicalin on iron-dependent lipid peroxidation (IDLPO) and the protective effect on pertussis bacilli-induced brain edema in rats. METHODS: In vitro, the inhibitory effect of baicalin and deferoxamine (DFX) was studied at different concentrations on IDLPO in brain homogenates of Sprague-Dawley rats. In vivo, the protective effect was studied of baicalin and DFX on brain edema induced by injecting purtussis bacilli into the left internal carotid artery in rats. Forty Sprague-Dawley rats were randomly divided into four groups. Group 1 (NS, n = 10), as control; Group 2 (PB, n = 10), pertussis bacilli control; Group 3 (DFX, n = 10), PB + deferoxamine; and Group 4 (BAI, n = 10), PB + baicalin. RESULTS: The baicalin was possessed of an inhibitory effect on IDLPO in rat brain homogenate, moreover, the inhibitory potency of baicalin was approximately 20 times higher than DFX treated alone. In vivo, the increase in the content of water(CW) and Evans blue (EB) was attenuated in groups DFX and BAI (CW: 79.8 +/- 0.7)% and (79.5 +/- 0.6)%; (EB: 11 +/- 7 micrograms/g and 12 +/- 5 micrograms/g) as compared with the group PB (CW: 80.4 +/- 0.9%, EB: 24 +/- 10 micrograms/g). The content of iron was significantly lower than in the left hemispheres (injected side) in the group DFX and the group BAI (79 +/- 9 micrograms/g and 77 +/- 10 micrograms/g) than in the left hemispheres in the group PB(132 +/- 28 micrograms.d-1.g), P < 0.05. The activation of CuZn-SOD (L155 +/- 36 U/mg pr, R177 +/- 75 U/mg pr) was decreased in the group BP compared with the group NS(L 201 +/- 49 U/mg pr, R 249 +/- 49 U/mg pr), P < 0.05. The activation of CuZn-SOD was significantly higher in the left brain tissues in the groups DFX and BAI (222 +/- 49 U/mg pr and 230 +/- 45 U/mg pr, respectively) than in the left brain tissues of the group of PB (P < 0.05). CONCLUSION: The baicalin and DFX have a protective effects on the pertussis bacilli-induced brain edema in rats, and the protective effects are related to the inhibitory effect on IDLPO, chelating Fe2+, and activated CuZn SOD. PMID- 11038819 TI - [Distribution of K5, K10 in transgenic mouse with lac Z reporter gene]. AB - OBJECTIVE: To detect the distribution of K5, K10 in live mouse. METHODS: lacZ gene, which encodes beta-galactosidase, as a reporter gene, bovine K5, K10 as promoter, containing Sal I fragment, was inserted into the Sal I sites of PLGZ and PLBS-2, respectively. The constructs were microinjected into the mice fertilized eggs, and then PCR positive ones were kept. The next generations could be used for experiments. Frozen sections of transgenic mice skin, tail were stained with 0.1% X-gal solution at 37 degrees C overnight and counterstained with nuclear fast red. RESULT: In sguamous epidermis, K5 was distributed in basal and outer root sheath, K10 in suprabasal and inner root sheath. K5 and K10 were also distributed in some other organs except heart. No K5 was seen in the brain. CONCLUSION: lacZ transgenic animal is helpful in the study of gene mutation and treatment. The recognition of K5 and K10 distribution could be a primary step to probe the mechanism of some related diseases and cancers. PMID- 11038820 TI - [Study of alpha-fetoprotein]. PMID- 11038821 TI - [A model of nasopharyngeal carcinoma with spontaneous highly lymphatic metastasis in nude mice and its biological characteristics]. AB - OBJECTIVES: To establish a model of nasopharyngeal carcinoma with spontaneous highly lymphatic metastasis in nude mice and to study its biological characteristics. METHODS: The clone cells of nasopharyngeal carcinoma(NPC), CNE 2Z-H5, were inoculated into nude mice. The cancer cells of lymph node metastatic foci were transplanted into nude mice again when the metastasis of nude mice were observed. After repetition of this procedure for 9 cycles, the metastatic rate and the metastatic paths were observed in nude mice of every passage; The changes were also observed in proliferation ability, expression of growth factor and its receptor in vitro, proliferation index(PI), and tumor weight and doubling time of the transplantable tumor in every passage in vivo. RESULTS: The lymphatic metastatic rates were higher than 85%. The metastatic paths were single, the proliferation ability was increased in cancer cells of every passage in vivo and vitro after passaging cancer cells of lymphatic metastatic foci for 9 cycles in nude mice. CONCLUSIONS: A model of nasopharyngeal carcinoma with spontaneous highly lymphatic metastasis has been established in nude mice, and the possibilities of metastasis and proliferation are increased after progression of NPC cells in nude mice. There is a close relationship between the metastasis and proliferation in NPC. PMID- 11038822 TI - [Mechanism of CD44V6 in human colorectal carcinoma metastasis]. AB - OBJECTIVE: To study the mechanism of CD44V6 in human colorectal carcinoma (HCC) matastasis. METHODS: The effects of CD44V6 on HCC cell adhering to base membrane and vascular endothelium were investigated by human amniotic membrane invasion model and co-culturing HCC cell with vascular endothelium. The influence of CD44V6 variants on cytoskeleton and gap junctional intercellular communication (GJIC) was studied by immunofluorescent confocal 3-dimensional reconstruction and fluorescence recovery after photobleaching technique. RESULTS: CD44V6 antibody can decrease the ability of HCC cells to adhere to amniotic membrane and vascular endothelium. It also can affect the distribution, polymerization and depolymerization of actin of HCC cells, but does not significantly affect GJIC of HCC cells. CONCLUSIONS: CD44V6 may play an important role in promoting HCC cells to adhere to vascular endothelium and base membranes and may affect the distribution, polymerization and depolymerization of actin in HCC cells and facilitate HCC cell metastasis. PMID- 11038823 TI - [Expression regulation of major late promoter tripartite leader sequence in adenovirus late infection]. AB - OBJECTIVE: To investigate the influence of adenovirus major late tripartite leader sequence on specific exportation of virus mRNA in the late phase of adenovirus infection. METHODS: We constructed a marked, human beta-actin minigene under the control of the glucocorticoid-inducible enhancer and promoter of mouse mammary tumor virus, and inserted it into the left end of the adenovirus type V genome. The promoter gene, designated MA, as well as of a sibling, which differed only in inclusion of a cDNA copy of the adenovirus major late tripartite leader sequence upstream of beta-actin sequences termed MtplA, in recombinant virus infected cells was strictly dependent on addition of deoxymethesone to the medium. Nuclear and cytoplasmic reporter RNA species were compared by Northern blotting, primer extension and pulse-chase assay. RESULTS: The high concentrations of newly-synthesized RNA were accumulated in cytoplasm when tripartite leader sequence was present in reporter RNA, despite the equal rates of transcription of the two reporter genes. Nuclear and cytoplasmic reporter RNA species compared by Northern blotting, primer extension, and pulse-chase provided no evidence for altered processing induced by tripartite leader sequence. CONCLUSION: The tripartite leader sequence, known to facilitate translation of mRNA during the late phase of adenovirus infection, can also modulate mRNA export from the nucleus. PMID- 11038825 TI - [Apoptosis in multiple organs of rats in early stage of polytrauma combined with shock]. AB - OBJECTIVE: To discuss Apoptosis in multiple organs of rats in early stage of polytrauma combined with shock. METHODS: DNA agarose gel electrophoresis, in situ end labeling (ISEL), light microscope and electron microscope were used and DNA fragmentation percentage (ap%) was detected. RESULTS: The special ladder pattern for apoptosis was seen in thymus, spleen, liver, lung and intestine, but not in heart, kidney and brain. At 6 hours after resuscitation, the ap% of thymus, spleen, liver, lung and intestine increased together with the severity of trauma. In six-site trauma combined with hemorrhagic shock group, the ap% of these five organs increased significantly at 1 hour after resuscitation and most significantly at 3 hours. At this point, the ap% of spleen, liver, lung and intestine reached peak, and declined gradually afterward. But the ap% of thymus continued to increase after 3 hours and kept stable from 6 hours to 24 hours ISEL showed that there were positive responses of different degrees in these eight organs. It was feand Morphologically most apoptotic cells in the thymus were positioned in the cortex, and those in spleen in the growing center of white pulp, and those in liver in the border area of hepatic lobule and portal area, and apoptosis of multiple kinds of cells including alveolar epithelial cells, vascular endothelial cells and polymorphonuclear neutrophils was induced in lung, and intestinal apoptotic cells laid in the epithelium and lamina propria of mucosa. CONCLUSION: Apoptosis was really induced in thymus, spleen, liver, lung and intestine in early stage of polytrauma combined with shock, which may play a role in early organ injury and late multiple organ failure. PMID- 11038824 TI - [Angiotensin II type 2 receptor inhibits type 1 receptor-induced cell proliferation and fibronectin release in mouse fibroblasts]. AB - OBJECTIVE: To investigate the effects of angiotensin II type 2 receptor (AT2) on the cell proliferation and fibronectin release induced by angiotensin II type 1 receptor (AT1). METHOD: Fibroblats were cultured from embryonic skin of AT2 receptor gene knocked-out mice (AT2-/-) and wild-type mice (AT2+/+) after 16 days of gestation, respectively. Cell proliferation and fibronectin release were examined in the fibroblasts treated with 10(-8) mol/L angiotensin II. RESULTS: Cell proliferation rate was increased more markedly in the AT2-/- fibroblasts than in the AT2+/+ fibroblasts after treament with angiotensin II (0.462 +/- 0.026 VS 0.389 +/- 0.021, P < 0.01). Also, fibronectin release and its gene expression were significantly enhanced in the AT2-/- fibroblasts as compared with the AT2+/+ fibroblasts when exposed to angiotensin II (34.1 +/- 4.1)% vs (16.2 +/ 2.3)%, P < 0.01). CONCLUSION: The AT2 receptor can inhibit the cell proliferation and fibronectin release induced by the AT1 receptor. PMID- 11038826 TI - [The effect of PMN adhesion mediated by CD11b/CD18 on the increasing permeability of microvascular endothelial monolayer after severe burn injury]. AB - OBJECTIVE: To detect the effect of burn-activated PMN adhesion and its adhesion molecule CD11b/CD18 on microvascular endothelial permeability using an experimental model of endothelial monolayer on polycarbonate microporous filters. METHODS: An experimental model for in vitro study of endothelal monolayer for permeability analysis was established. Seven groups were divided into according to the treatment of microvascular endothelial monolayer. Fluid filtration coeffecient(Kf) and albumin reflection coeffecient(delta) were measured after endothelial monolayer was perfused with albumin labelled by FITC. RESULTS: Burn activated PMN could increase the level of fluid filtration coeffecient(Kf) and decrease the albumin reflection coeffecient(delta). Monoclonal antibody sealing off CD11b/CD18 on PMN provented the change of delta induced by burn-activated PMN. Another microporous filter interposed between PMNs and endothelial monolayer corrected the changes of Kf and delta. CONCLUSION: The permeability enhancing effect of PMNs may be attributed mainly to the PMN-EC adhesion mediated by CD11b/CD18. Blocking the PMN-EC over-adhesion in moderation may be helpful in reducing the lung injury due to severe burn injury. PMID- 11038827 TI - [Vascular endothelial growth gene therapy for narrowing of rabbit iliac artery]. AB - OBJECTIVE: To study the mechanism of renarrowing and vascular endothelial growth factor (VEGF) gene therapy to prevent the restenosis caused by balloon angioplasty. METHODS: According to the special function of VEGF that promepts the proliferation of endothelial cell (EC), we constituted an eukaryotic expression vector of pcDNA3/VEGF, and added the pcDNA3/VEGF to the surface of polylysine balloon. We examined the results of the test by in situ blot, eletromagnetic blood flow mechine, scanning eletromicroscope, and light microscope. RESULTS: There was higher expression of VEGF mRNA and iliac artery intima in the VEGF gene group. The extent of endothelialization was larger, the ratio of intima to media (I/M) was smaller, and the blood flow was more sufficient than in the vector control group. CONCLUSION: VEGF gene local delivery could stimulate the proliferation of EC, accelerate the endothelialization of injury-artery, and inhibite the proliferation of smooth muscle cell (SMC). PMID- 11038828 TI - [Free fatty acids promoting PTP1B expression in rat skeletal muscle and hepatic cells]. AB - OBJECTIVE: To determine whether that free fatty acids (FFAs) impair glucose metabolism is associated with altered insulin signaling system. METHODS: After skeletal muscle and hepatic cells were incubated with palmitate (0.25 mmol/L) or oleate (0.125 mmol/L) for 6, 12 or 24 hours, the protein abundance of SH-PTP2 and PTP1B was assessed by western blot. RESULTS: SH-PTP2 protein levels showed no significant change in both fatty acids treated muscle and hepatic cells at all time points. The PTP1B was significantly elevated in both liver and muscle cells incubated with FFAs at 6, 12 and 24 hours. CONCLUSIONS: FFAs promote the expression of PTP1B in rat skeletal muscle and liver cells, and the elevated PTP1B may mediate the insulin resistance induced by FFAs. PMID- 11038829 TI - [Diagnosis and treatment of skull base metastasis]. AB - OBJECTIVE: To study the diagnosis and surgical treatment of skull base metastases. METHODS: Clinical and radiological features, operative technique and postoperative complications of 15 cases of skull base metastases were discussed. RESULTS: Total resection was achieved in 13 cases, and subtotal resection in 2. 7 tumors were located in the anterior skull base. The main complication was CSF leakage. CONCLUSION: The preoperative diagnosis of skull base metastases is difficult, radical resection is effective, and postoperative complications are not uncommon. PMID- 11038830 TI - [DNA typing for HLA-A, B antigens by polymerase chain reaction with sequence specific primers and clinical application]. AB - OBJECTIVE: To establish DNA typing for HLA-A, B antigens in Chinese by polymerase chain reaction with sequence-specific primers (PCR-SSP). METHODS: DNA samples were obtained from 178 unrelated donors and 167 kidney recipients. An additional panel of 62 standard DNAs that were typed by UCLA tissue typing lab in USA. A rapid genotyping for HLA-I class (A, B antigens) by PCR-SSP was set up by designed and synthesized 81 specific primers and 1 pair of internal control primer, combining in 61 one-step reactions (20 PCR reactions for A alleles, 41 PCR reactions for B alleles). RESULTS: HLA-A, B alleles were successfully typed in 345 clinical samples and 62 standard DNAs by PCR-SSP technique. No false positive or false negative typing results were obtained. Reproducibility was 100% in 40 samples. The overall time of DNA typing was 5 hours. The typing results were consistent with those of UCLA tissue typing lab. Nineteen alleles of HLA-A and 41 HLA-B alleles were accurately distinguished. Thirteen HLA-A alleles and thirty-two HLA-B alleles in Chinese were practically typed. CONCLUSION: DNA typing for HLA-I class (A, B antigens) by PCR-SSP has proved to be a technique of high-resolution, high-specificity, well-reproducibility, and more suitable for clinical application than serology. PMID- 11038831 TI - [The pharmacokinetic study of desflurane, sevoflurane, isoflurane and enflurane in general anesthesia]. AB - OBJECTIVE: To compare pharmacokinetics of desflurane, sevoflurane, isoflurane and enflurane in general anesthesia. METHODS: 40 patients scheduled for abdominal hysterectomy under general anesthesia were randomly divided into desflurane(D), sevoflurane(S), isofluane(I) and enflurane(E) groups. After induction of anesthesia and endotracheal intubation, desired fraction (Fd) of desflurane(6%), sevoflurane(2%), isoflurane (1.15%) and enflurane(1.7%) in oxygen and nitrous oxide(1:2) were inhaled in D, S, I and E groups, respectively. The fractional end tidal alveolar concentration (Fa) was adjusted to 1MAC during the maintenance of anesthsia. Fa and the fractional inspired concentration of inhaled anesthetics (Fi) were monitored continuously. During operation, fentanyl was infused continuously and pancuronium was injected intermittently. RESULTS: After the beginning of inhalational anesthesia, the time required for Fa/Fi = 1:2 and Fa = 1MAC in D and S groups was significantly shorter than that in E and I groups. The rates of Fa/Fi in D and S groups were significantly higher than those in E and I groups during the maintenance of anesthesia, so were those of Fa/Fd. After cessation of inhalational anesthesia, the time required for Fa equaled to 50% of Fa0(the last Fa during stoping administration of the inhalational anesthesia) in D group was significantly faster than that in the other three groups. CONCLUSIONS: The rates of desflurane wash-in and wash-out are faster than those of other inhaled anesthetics. The depth of anesthesia is easy to control when desflurane is used in general anesthesia. PMID- 11038832 TI - [Changes of erythrocyte and platelet membrane lipid pattern in different subtypes of dementia]. AB - OBJECTIVE: To investigate the changes of platelet and erythrocyte membrane lipids and phospholipid composition in different types of dementia. METHODS: There were 19 patients with Alzheimer's disease, 25 patients with Binswanger's disease, 23 patients with multi-infarct dementia, 11 patients with single cortical infarct dementia, 7 patients with vascular dementia of haemodynamic type, 18 patients with dementia following Parkinson's disease, and 25 senile controls. Platelet and erythrocyte membrane cholesterol, total phospholipids and individual phospholipids were quantified. RESULTS: When compared with senile controls, the decrease of total phospholipids, phosphatidylcholine and phosphatidylethanolamine and the increase of cholesterol and cholesterol/total phospholipids ratio on the erythrocyte and platelet membrane were found in patients with Binswanger's disease and in those with multi-infarct dementia. The increase of erythrocyte and platelet membrane cholesterol, cholesterol/phospholipids ratio was found, but there was no significant change in membrane total phospholipids and phospholipid composition in patients with single cortical infarct dementia. CONCLUSION: Metabolic disorders of platelet and erythrocyte membrane phospholipids were noted in patients with Binswanger's disease. These disorders were related to white matter low attenuation. PMID- 11038833 TI - [Sulfonylurea receptor gene polymorphism is associated with non-insulin dependent diabetes mellitus in Chinese population]. AB - OBJECTIVE: To test the role of Sulfonylurea receptor gene(SUR) in the pathogenesis of NIDDM in Chinese population. METHODS: We studied the polymorphisms of the SUR gene in intron 24 and exon 22 by polymerase chain reaction(PCR) and appropriate restriction enzyme (PCR-RFLP) in 86 NIDDM patients with at least two first degree diabetic relatives and 148 normal control subjects. RESULTS: The frequency of "c" allele of intron 24 in NIDDM patients was significantly increased as compared with that in the control subjects (68.02% vs. 55.41, P = 0.007), and the frequency of the "cc" genotype of intron 24 in the NIDDM group was also significantly higher than that in the control group (41.86% vs. 27.7%, P = 0.013, OR = 4.39, CI: 1.52-12.66). The polymorphism of exon 22 described in the Caucasian population was not detected. CONCLUSION: The association of the polymorphism of SUR gene with NIDDM in different races suggests that the SUR gene or nearby gene may play an important role in the genetic susceptibility of NIDDM. PMID- 11038834 TI - [The pressure of amniotic cavity during pregnancy and treatment of unruptured oligohydramnios with antepartum amnioinfusion]. AB - OBJECTIVES: To measure the pressure of the amniotic cavity during pregnancy and to study the effects of antepartum amnioinfusion on oligohydramnios with intact membranes. METHOD: We used an improved three-way switch apparatus to measure 110 women during pregnancy, and treated 97 oligohydramnios with antepartum amnioinfusion or intravenous infusion as controls. RESULTS: The pressure of the amniotic cavity was 1.69 +/- 0.23 kPa, which was close to the normal value in pregnancy. In oligohydramnios, it was 1.1 +/- 0.3 kPa, and in polyhydramnios 3.2 +/- 0.3 kPa(P < 0.01-0.001). When 300 ml fluid was amnioinfused, the amniotic fluid index (AFI) increased by 5.0 +/- 1.8 cm, the pressure of amniotic cavity increased by 0.18 +/- 0.2/kPa and the incidence of vaginal delivery was 87.9%, whereas that of the control group was 12.1%. There were no fetal distress, asphyxia neonatorum, amnionitis or ruptured membrane. CONCLUSIONS: The pressure of the amniotic cavity during normal pregnancy was stable, but changed with AFI. Antepartum amnioinfusion for unruptured oligohydramnios resulted in a marked increase in incidence of vaginal delivery, a decreased ratio of cesarean section and a reduction of perinatal morbidity. These differences are highly significant. PMID- 11038835 TI - [The effects of diltiazem on pulmonary hemodynamics and oxygenation in copd induced pulmonary hypertension]. AB - OBJECTIVE: To study hemodynamic the effects of intravenous diltiazem on COPD induced pulmonary hypertention. METHODS: 11 cases in exacerbation stage were chosen to study. Swan-Ganz catheterization were taken for hemodynamic monitoring. RESULTS: After the administration of diltiazem, pulmonary vascular resistance, pulmonary pressure, right artrial pressure and heart rate decreased significantly (the extents decreased were 14.0 kPa.s-1.L-1, 0.62-1.27 kPa, 0.27-0.40 kPa and 25 30 beats/min respectively, P < 0.05); Cardiac output and stroke volume increased identically (the extents increased were 0.3-0.6 L.min-1.m-2 and 5-9 ml.beat-1.m-2 respectively, P < 0.05); No significant changes found in Qs/Qt, PaO2 and systemic blood pressure. CONCLUSION: Our study showed that diltiazem produces beneficial pulmonary hemodynamic effects without oxygenation compromise in COPD cases and has a strong indication in those who complicated with superventricular tachycardia. PMID- 11038836 TI - [The evaluation of electron microscopy in the pathological diagnosis of renal biopsies]. AB - OBJECTIVE: To evaluate the use of electron microscopy(EM) in pathological diagnosis of renal biopsies. METHOD: 777 cases of renal biopsy in Department of Nephrology, First Hospital of Beijing Medical University from Jan 1995 to Jun 1997 were studied. We compared the preliminary diagnosis by light microscopy(LM) and immuno-fluorescence(IF) with the final diagnosis by electron microscopy(EM). The use of EM in the diagnosis was divided into three parts. EM was needed to make a correct final diagnosis, provided confirmatory data or additional information relevant to the diagnosis and was complementary to LM and IF. RESULTS: Among 777 cases of renal biopsy, EM was needed to make a correct final diagnosis in 18.5%, and provided important referential information in 13.5%. CONCLUSION: EM provided useful diagnostic information in about one third cases of renal biopsy. PMID- 11038837 TI - [Lung volume reduction surgery for pulmonary emphysema caused by chronic obstructive pulmonary disease]. PMID- 11038838 TI - [Progress in the gene research on hereditary breast cancer-ovarian cancer]. PMID- 11038839 TI - [Progress in the study of cardiology in China]. PMID- 11038840 TI - [Progress in the study of respiratory diseases in China]. PMID- 11038841 TI - [Progress in the study od renal diseases in China]. PMID- 11038842 TI - [Progress in the study od gastrointestinal diseases in China]. PMID- 11038843 TI - [Progress in the study of hematologic diseases in China]. PMID- 11038844 TI - [Progress in the study of rheumatic diseases in China]. PMID- 11038845 TI - [Progress in the study of infectious diseases in China]. PMID- 11038846 TI - [Progress in the study of tuberculosis in China]. PMID- 11038847 TI - [Progress in the study of neurologic diseases in China]. PMID- 11038848 TI - [Progress in the study of geriatrics in China]. PMID- 11038849 TI - [Progress in the study of pediatrics in China]. PMID- 11038850 TI - [Progress in the study of emergencies in China]. PMID- 11038851 TI - [Progress in the study of cardiovascular surgery in China]. PMID- 11038852 TI - [Progress in the study of general surgery in China]. PMID- 11038853 TI - [Progress in the study of orthopedics in China]. PMID- 11038854 TI - [Progress in the study of urinary surgery in China]. PMID- 11038855 TI - [Progress in the study of neurosurgery China]. PMID- 11038856 TI - [Progress in the study of burn surgery in China]. PMID- 11038858 TI - [Progress in the study of microsurgery in China]. PMID- 11038857 TI - [Progress in the study of organ transplantation in China]. PMID- 11038859 TI - [Progress in the study of pediatric surgery in China]. PMID- 11038860 TI - [Progress in the study of gynecology in China]. PMID- 11038861 TI - [Progress in the study of perinatology in China]. PMID- 11038862 TI - [Progress in the study of anesthesiology in China]. PMID- 11038863 TI - [Progress in the study of oral medicine in China]. PMID- 11038864 TI - [Progress in the study of ophthalmology in China]. PMID- 11038865 TI - [Progress in the study of dermatologic and gynecologic diseases in China]. PMID- 11038866 TI - [Progress in the study of family planning in China]. PMID- 11038867 TI - [Progress in the study of ultrasonography in China]. PMID- 11038868 TI - [Progress in the study of radiology in China]. PMID- 11038869 TI - [Progress in the study of radiation medicine and radiation protection in China]. PMID- 11038870 TI - [Progress in the study of laboratory diagnosis in China]. PMID- 11038871 TI - [Progress in the study of nuclear medicine in China]. PMID- 11038872 TI - [Progress in the study of laser medicine in China]. PMID- 11038873 TI - [Progress in the study of clinical pharmacology in China]. PMID- 11038874 TI - [Progress in the study of pathology in China]. PMID- 11038875 TI - [Progress in the study of cytobiology in China]. PMID- 11038876 TI - [Progress in the study of microcirculation in China]. PMID- 11038877 TI - [Progress in the study of clinical epidemiology in China]. PMID- 11038878 TI - [Progress in the study of psychiatry in China]. PMID- 11038879 TI - [Progress in the study of physical medicine and rehabilitation in China]. PMID- 11038880 TI - [Analysis of malate dehydrogenase isozymes in Taihe negro fowl by thin-layer isoelectric focusing polyacrylamide gel electrophoresis]. AB - The isozyme patterns of malate dehydrogenase (MDH) from six tissues (liver, kidney, eye, muscle, brain and heart) of the Taihe negro fowl (Gallus gallus domesticus Brission) were analysed by thin-layer isoelectric focusing polyacrylamide gel electrophoresis. It was found that there were twelve MDH isozyme bands in the tissues of the fowl, and these isozymes displayed a tissue specific distribution. PMID- 11038881 TI - [Identification of realgar and arsenopyrite and their sublimates]. AB - In this paper, two traditional Chinese medicinal minerals realgar and arsenopyrite, as well as their sublimates are identified in their nature, physical and optical properties, chemical compositions and trace elements. The result has shown that there are marked differences among realgar, arsenopyrite and their sublimates, and it should be particularly noted that the difference between the two sublimates will certainly affect the quality of the two minerals that are used as poisonous drug in China. PMID- 11038882 TI - [Investigation on new resources of medicinal plant Scrophulariaceae in Sichuan province]. AB - In this paper, 3 species of Scrophulariaceae which are found in Sichuan for the first time and 15 species of the same plant with their medical effects are reported. All of these species are described in detail including their distributions, environments and functions. PMID- 11038883 TI - [Isoesterase assay over the course of dormancy relieving with low temperature in the seed of Fritillaria thunbergii Miq. by electrophoresis]. AB - In order to study the physiological basis of dormancy relieving with low temperature in the seed of Frit-illaria thunbergii isoesterase patterns were assaved by means of electrophoresis over the courses of dormancy relieving at 8 10 degrees C and 3-5 degrees C. The changes of isoesterase patterns are similar at the two different temperatures. Over the courses of dormancy relieving, some new bands are detected and some original bands are enhanced. These results show that the physiological basis may be similar over the courses of dormancy relieving at 8-10 degrees C and 3-5 degrees C. PMID- 11038884 TI - [Comparative studies on pharmacological and toxic actions of raw fructus Psoraleae and its salt-baked and improved salt-baked products]. AB - The pharmacological and toxic actions of raw Fructus Psoraleae and its salt baked, and improved salt-baked products were comparatively studied. The leucogenic action of the improved salt-baked product was more effective than that of the original salt-baked one. But no significant difference in antidiarrheal or antiandrogenoid effect was found between the two salt-baked products. The toxicity(LD50 and the injury on kidney) of the improved salt-baked product was lower than that of the salt-baked. one. The results indicate that this improved processing method would contribute to increasing the efficacy and decreasing the toxicity of Fructus Psoraleae. PMID- 11038885 TI - [Effect of processing on reducing the toxicity in sulfur]. AB - The content of arsenic in sulfur before and after processing was determined. The result shows that the content in raw materials is 8-15 times higher than that of the processed product, which implies that the toxicity of sulfur is obviously reduced by processing. PMID- 11038886 TI - [Comparison of 4 extraction methods of chemical constituents in medicinal tea sishen chaji]. AB - Four extraction method for the medicinal tea Sishen Chaji, were compared with psoralen, schizandrin B and evodiamine taken as indexes. The result shows that the total contents of the three compounds decrease progressively in the following order: semi-bionic extraction, semi-bionic extraction by precipitation with alcohol, extraction with water, and extraction with water and precipitation with alcohol. PMID- 11038887 TI - [Chemical constituents of Hemsleya graciliflora (Harms) Cogn]. AB - Nine compounds were isolated from the fruits of Hemsleya graciliflora. IIThey were identified as- 25-acetate of dihydrocucurbitacin F(I), dihydrocucurbitacin F(II), cucurbitacin F-25-acetate(III), sitostery1-3-O-beta-D-glucoside-6'-O palmitate(IV), daucosterol(V), beta-sitosterol(VI), palmitic acid(VII), octodecyl acohol(VIII) and hexadecanol(IX) on the basis of physical and chemical properties as well as spectral data. The compound(IV) is isolated from genus Hemsleya for the first time. PMID- 11038888 TI - [Chemical constituents of the stems of Erycibe schmidtii Craib]. AB - Seven compounds were isolated from the stems of Erycibe schmidtii for the first time. They were identified as hexadecanoic acid, beta-sitosterol, scopoletin, chlorogenic acid, daucosterel, scopolin and an alkaloid-baogongteng C. PMID- 11038889 TI - [GC-MS analysis of constituents of essential oil from twigs and leaves of Tamarix chinensis Lour]. AB - The essential oil from the dried twigs and leaves of Tamarix chinensis was analyzed by GC-MS qualitatively and GC quantitatively. Sixty-three constituents were identified, which make up 81.76% of the oil. Hexadecanoic acid (22.22%) was the main constituent. PMID- 11038890 TI - [Antilipid peroxidation and antiradiative action of glycosides in herba Cistanches]. AB - The results showed that oral administration of GCs could markedly increase the activity of SOD in red blood cells as well as the contents of nucleic acid (DNA and RNA) in liver and kidney, decrease the contents of MDA in serum, markedly increase the activity of SOD in red blood cells as well as the contents of nucleic acid in the spleen of mice radio-induced by 60Co, and also lower the contents of MDA in liver. The results suggest that the protective effect of GCs on nucleic acid and antiradiative action may be related to its antilipid peroxidation. PMID- 11038891 TI - [Immuno-enhancing effect of gebie oral liquid on mice]. AB - Gebie Oral Liquid can increase the weight of thymus and spleen of the mouse, damaged by prednisone and cytoxan, enhance the phagocytosis of monocytes and DHT, and promote the blastogenesis of splenic lymphocytes as well as the activity of NK cells. PMID- 11038892 TI - [A study on the action of tang niao kang decoction]. AB - Experiments have verified that Tang Niao Karg (a preparation from Chinese medicinal herbs) taken orally does not decrease the blood sugar of empty stomach in mice, but markedly helps lower high levels of blood sugar and lipid induced by epinephrine and glucose in mice. For rat models of diabetes induced by 4-o pyrincidine Tang Niao Kang can significantly lower the content of blood sugar, raise the insulin level of blood serum, and enhance the tolerance to sugar. PMID- 11038893 TI - [Inhibitory effect of 7 agents on bovine testis aldose reductase]. AB - The inhibitory effects of seven agents on aldose reductase were studied by enzymatic kinetics using the purified enzyme from bovine testis, sorbinil, tolrestat, silybin and berberini were found to exhibit a marked enzyme-inhibitory effect with Ki values of 1.08 mumol/L, 0.12 mumol/L, 3.5 micrograms/ml, and 2.7 micrograms/ml respectively. The established assay system described in this paper may be useful in screening aldose reductase inhibitors. PMID- 11038894 TI - [Commercial identification and resources survey on Chinese drug radix Ranunculi ternati]. AB - Based on the collection of commercial products field investigation in distribution areas and main production places, and consultation of over 200 specimens the identification of traditional Chinese drug Maozhaocao was carried out. The results show that the present source of commercial "Maozhaocao" derives from 2 species of genus Ranunculus. The main species is Ranunculus ternatus, while R. polii is used only in a few regions. A resources survey of the drug was also conducted. PMID- 11038895 TI - [Continuity and changes of breeds of fupenzi]. AB - Based on textual researches, it has been confirmed that Fupenzi first appeared in Han Dynasty. Both Fupenzi and Penglei had the same origin, but split up during Jin Dynasty into two different breeds. Through ages Fupenzi has been taken as the fruits of the plant Rubus of Rosaceae species. Among them, R. idaeus, R. corchorifolius and R. coreanus have long been in use. R. chingii is the new breed of modern Fupenzi. PMID- 11038896 TI - [Changes of soluble protein contents in Fritillaria thunbergii Miq. over the course of dormancy relieving with low temperature]. AB - The changes of soluble protein contents in bud, adaxial cortex and inner tissue in the scale of Fritillaria thunbergii were assayed over the course of dormancy relieving with low temperature. There was a peak in each of the three diagrams at 25 days or 35 days treated with low temperature. All of the soluble protein contents of bud, adaxial cortex and inner tissue in scale tended to increased in the stage before dormancy relieving. PMID- 11038897 TI - [Culture techniques for high yield and high benefit of wild Polygonatum sibiricum Redoute]. AB - Through comprehensive studies on the artificial cultivation of wild Polygonatum sibiricum a complete set of culture technique for high yield and high benefit have been raised to guarantee a high yield of one thousand catties of grain perum and an increase in income of about 1000 yuan per mu per annum. PMID- 11038898 TI - [Catalpol content changes in Rehmannia glutinosa (Gaertn.) Libosch. under certain conditions]. AB - HPLC determination has shown that the content of catapol in Rehmannia glutinosa decreases markedly after being dried and processed. This change of content is related to acidic and alkaline conditions. But in boiling water or saccharide solution no marked changes have been observed. PMID- 11038899 TI - [Determination of myristicin and safrol in volatile oil of nutmeg and its processed products by HPLC]. AB - The contents of myristicin and safrol in the volatile oil of nutmeg and its processed products were determined by HPLC. This method is fast, accurate and gives good resolution. It has also been confirmed that myristicin tends to decrease in varying degrees after the nutmeg has been processed. PMID- 11038900 TI - [Comparison of two extraction methods for maxingshigan decoction]. AB - The semi-bionic extraction and the extraction with water, two methods for the Maxingshigan Decoction, were compared. The results show that the semi-bionic extraction is superior to the extraction with water in the yield of ephedrine, hydrocyanic acid, glycyrrhetinic acid, calcium ion and extract. PMID- 11038901 TI - [Chemical constituents of rhizoma Pinelliae pedatisecta]. AB - Five compounds were isolated from the alkaloid extract of Rhizoma Pinelliae Pedatisecta and their structures were determined as pedatisectine F(I), hypoxanthine (II), erythritol (III), uridine (IV) and pedatisectine G (V), of which I and V are new compounds, while II, III and IV were found in this plant for the first time. PMID- 11038902 TI - [Fat soluble constituents of the leaves of Vaccinium bracteatum Thunb]. AB - Four compounds were isolated from the fat soluble fraction of the leaves of Vaccinium bracteatum and identified as friedelin (I), epifriedelinol (II), beta sitosterol(III) and ursolic acid(IV) by IR, NMR and MS. Compound III and IV are isolated from the leaves of this plant for the first time. PMID- 11038903 TI - [GS-MS analysis of essential oils from five species of Asarum]. AB - This paper reports the result of GS-MS analysis of the essential oils from five species of Asarum, namely, A. heterotropoides var. mandshuricum (cultivated), A. sieboldii (cultivated), A. caudigerellum (from Sichuan), A. sieboldii(from Shandong) and A. sieboldii(wild). Ninety-two constituents were detected, of which 73 compounds were identified. PMID- 11038904 TI - [Effects of total flavones of fructus Hippophae (TFH) on cardiac function and hemodynamics of anesthetized open-chest dogs with acute heart failure]. AB - This study was carried out employing the model of dog with acute heart failure induced by phenobarbital natricum. It was shown that i.v. TFH 4.8 and 9.7 mg/kg could significantly increase CO, CI, + LVdP/dtmax and LVSP; shorten R-dP/dtmax in 9.7 mg/kg; raise - LVdP/dtmax, reduce LVEDP and T value; decrease MVO2I and TPVR; MAP and HR were not changed significantly. The results suggest that i.v. TFH can strengthen cardiac pump function and myocardial contractility in canine with heart failure; significantly improve myocardial diastolic function and hemodynamic performance; markedly decrease myocardia-used oxygen index and total peripheral vessel resistance, thus proving that TFH is good for heart failure. PMID- 11038905 TI - [Therapeutic effect of radix Astragali on hypoxia pulmonary hypertension in rats]. AB - Radix Astragali could inhibit the type III collagen deposition, smooth muscle cell proliferation in IAPA, and increase the tolerance of myocardium to hypoxia in rats. PMID- 11038906 TI - [Pharmacological effects of zhenzhu jingmu oral liquid]. AB - Zhenzhu Jingmu Oral Liquid (ZJOL) is an aqueous solution extracted from the serum of spermary and ovary of Pinctada martensii. Taurine, an effective chemical component, has been isolated from ZJOL. It has been shown that ZJOL given orally can produce an excitative constrictive effect on the uterus of guinea pigs in vivo, prolong the dissolving time of euglobulin in rabbits, and shorten the bleeding time as well as the blood clotting time in mice. In higher dosages, ZJOL can cause a sedation in mice. PMID- 11038907 TI - [Progress in the clinical uses of flos Carthami]. PMID- 11038908 TI - [A review of studies on the resources of Chinese herbal drugs in recent years]. PMID- 11038909 TI - [Survey of botanical origin of tongcao (medulla Tetrapanacis) and identification of its commercial products]. AB - Through the investigation on botanical origin, output and sales of Togcao in producing areas, it has been proved that there are twenty-two species of six families used as Tongcao. The medicinal parts are the pith of stems or petioles. The provinces featuring more species and larger output of Tongcao are Sichuan, Yunnan, Guizhou, Guangxi, Hunan and Shaanxi. A hundred and two pieces of commercial samples collected from twenty-six provinces in China, Hongkong area, Japan, Malaysia, Thailand, Singapore and Republic of Korea have been identified. The result shows that both Xiaotongcao and Datongcao are called by the same name Tongcao. The main species is Xiaotongcao, which takes a proportion of 70% in Tongcao. And the Tongcao(Tetrapanax papyriferus, taking a proportion of 20%) as recorded in the Chinese Pharmacopoeia (1995 edition) is seldom used. PMID- 11038910 TI - [Damage-loss relationships and integrated control of 3rd instar larvae of Actias selene nigpoana Felder in planting area of Cornus officinalis Sieb. et Zucc]. AB - In the present paper, the damage-loss model of Actias selene ningpoana to Cornus officinalis was tested and the results indicated that the yield loss rates obeyed the equation Y = 100 - EXP(4.6042 - 0.0315X). The economic threshold of the 2hd and 3rd instar larvae of A. selene nigpoana was then determined as 22 and 8 insects per tree respecitively. Suggestions for integrated control have been made based on the research of activities of A. selene ningpoana in the forest. PMID- 11038911 TI - [Diastase and reducing sugar in Fritillaria thunbergii Miq. during the process of dormancy relieving with low temperature]. AB - The changes of diastase's activity and reducing sugar's content in Fritillaria thunbergii were studied during the process of relieving dormancy with low temperature. The results showed that the metabolism in bud and scale was changed, the diastase's activity and reducing sugar's content of the adaxial epiderm were higher than those of the inner store tissue in the scale, but were approximate to those of the bud. PMID- 11038912 TI - [Outward character and inner quality of Bombyx Batryticatus, Carapax Trionycis, Squama Manitis and endothelium corneum Gigeriae Galli after frying]. AB - According to the requirement for outward character in processing traditional Chinese medicines, Bombyx Batryticatus, Carapax Trionycis, Squama Manitis and Endothelium Corneum Gigeriae Galli, which are four kinds of often used traditional Chinese animal medicines, were fried, the contents of amino acids in each kind were determined, and investigate the inner quality of each kind of the processed product was investigated with reference to other standards. PMID- 11038913 TI - [Determination of eugenol, methyleugenol and methylisoeugenol in volatile oil of differently processed nutmeg]. AB - A HPLC method for the determination of eugenol, methyleugenol and methylisoeugenol in the volatile oil of differently processed natmeg has been used. The result has shown that the content of eugenol is only sliahtly changed before and after processing, but that of methyleugenol and methylisoeugenol has obviously increased. PMID- 11038914 TI - [New method for preparing oridonin by column chromatography]. AB - Compared with the traditional methods, the separation and preparation of oridonin by column chromatography have the advantages of fast speed, harmlessness and non pollution of solvent, safety, economy and easily obtained, etc. After re crystallizing the oridonin content is determined by TLC scanning to be as high as 97.03%. PMID- 11038915 TI - [Chemical constituents of Bulgaria inquinans (Fries)]. AB - Three compounds were isolated from Bulgaria inquinans. They were identified as ergosterol, galacitiol and ethanedioic acid. PMID- 11038916 TI - [Chemical constituents of the stems and leaves of Trigonella foenum-graecum L]. AB - Two compounds were isolated from the leaves and stems of Trigonella foenum graecum, and on the basis of spectral analysis, their structures were elucidated as gamma-schizandrin and scopoletin. They were isolated from T. foenum-grecum for the first time. PMID- 11038917 TI - [Chemical studies on peptidepolysaccharides of Ganoderma lucidum (W. Curt. Fr.) Karst]. AB - Seven homogeneous peptidepolysaccharides have been obtained from the fruit-body of Ganoderma lucidum. The main constituents are TGLP-2, TGLP-3, TGLP-6 and TGLP 7, whose molecular weights are 20.9 x 10(4), 4.5 x 10(4), 3.2 x 10(4) and 10 x 10(4) respectively. TGLP-2 is a heteroglycan peptide, TGLP-3 and TGLP-6 are glycan peptides and TGLP-7 is a galactan peptide. PMID- 11038918 TI - [GC-MS analysis of constituents of essential oils from stems of Ephedra sinica Stapf, E. intermedia Schrenk et C.A. Mey. and E. equisetina Bge]. AB - The essential oils from the dried stems of Ephedra sinica, E. intermedia and E. equisetina were analyzed by GC-MS qualitatively and GC quantitatively. One hundred and twenty-seven constituents were identified, l-alpha-terpineol (31.64%) in E. sinica, 1,4-cineole (12.80%) in E. intermedia and hexadecanoic acid (26.22%) in E. equisetina being the main constituents. PMID- 11038919 TI - [Effects of yizhi pills on memory, superoxide dismutase and malondialdehyde of brain and immunity in mice]. AB - Passive avoidance tests have shown that Yizhi Pills (YZ) markedly improve the memory of normal mice at a dose of 100 mg/kg after oral administration for fifteen days, and significantly reverse the scopolamine, NaNO2 and EtOH-induced disruptions of memory retention in mice at doses of 100, 200, 500 mg/kg after oral administration for five days. In aged mice induced by D-galactose, YZ also significantly improve the impaired memory, increase the activity of SOD and decrease the content of MDA in brain. All these effects were observed at doses of 200, 500 mg/kg after oral administration for forty-one days. YZ significantly promote blood carbon particle clearance, enhance hemolysin antibody in immunodepressed mice induced by cyclophosphamide, and increase earswelling in immunodepressed mice induced by prednisolonum. PMID- 11038920 TI - [Comparison of pharmacological studies on ootheca Mantidis]. AB - The pharmacological effects of three species of Ootheca Mantidises were compared. The results indicate that Tenodera sinensis can increase the index of testis and thymus gland, and has an antidiuretic effect in mice; Statilia maculate can prolong the swimming time and ordinary pressing anoxia, increase the index of spleen and thymus gland, raise the temperature in mice, decrease the content of LPO in liver of the hypercholesteremia rats, and has an antidiuretic effect; Hierodula patellifera can increase the index of testis and thymus gland, raise the temperature in mice, and decrease the content of LPO in liver of the hypercholesteremia rets. The LD50 of the three species of Ootheca Mantidises are all over 320 g/kg. PMID- 11038921 TI - [Effects of icarrin on secretive function of cultured granulosa and adrenal cortical cells of rats]. AB - Icarrin was added to cultured cells and the hormones in culture fluid were determined after 3 hours of incubation. The results showed that icarrin (30-1000 micrograms/L) could promote the estradiol production of granulosa cells. The most effective concentration was 100 micrograms/L. Icarrin also exerted a stimulatory effect on corticosterone production at the highest dosage (1000 micrograms/L). PMID- 11038922 TI - [Extraction and sequencing of trace DNA from cornu Cervi pantotrichum]. AB - The trace DNA extraction method was used to extract DNA from the blood, hair, Cornu cervi pantotrichum, hairy antler of the sika deer, Cervus nippon, and penis of Bulbalus bublis and Asinus vulgaris. A 307 bp fragment of mitochondrial DNA cytochrome b gene was amplified with primers L14841 and H15149 for these extractions. The purified PCR products were sequenced by the dideoxy method. DNA sequence obtained from the Cornu cervi pantotrichum is the same as that obtained from the fresh blood and hair of the sika deer, but the sequence of the so-called hairy antler is quite different, implying that it is not from the sika deer. In addition, the phylogenetic relationships among different species are congruent with the traditional known phylogeny. PMID- 11038923 TI - [Prospects of application of DNA molecular genetic marker to pharmacognostical identification]. AB - The theoretical foundation of application of DNA molecular genetic marker to pharmacognostical identification is summarized. The practical application of the marker is reviewed, the matters deserving attention during application are pointed out, and vistas of the application are spelled out. PMID- 11038924 TI - [Ontogeny of Curcuma longa L]. AB - Based on the research of growth of organs, accumulation of dry matter, distribution of photosynthetic production and displacement of growth centre, the growth of turmeric can be divided into three stages. There are 13 leaves in the life of turmeric. Leaf formation takes place rapidly to form 8 leaves in the seedling stage. The leaf area and increase of dry matter per day in roots and leaves are smaller or lower in the seedling stage. In the daughter rhizome formation stage, NAR reaches the maximum (3.54/m2.d), and the leaf area reaches the maximum(3302.9 cm2/plant, LAI 4.95). Before the daughter rhizome formation stage, more than 75% of dry matters are distributed in leaves, and in the daughter rhizome formation stage, 50%-75% of dry matters are distrbuted in leaves. In the late growing period, more than 40% of dry matters are distributed in the daughter rhizome. The growth centre of turmeric changes two times in life. When CGRr is equal to CGRt, the crossing of CGRt and CGRr curves may be regarded as the evidence of the transfer of growth centre from leaves to daughter rhizome. PMID- 11038925 TI - [Optimization of technical parameters for processing radix Aconiti coreani]. AB - Based on the determination of guanfu A, hypaconitine, and total alkaloids, along with the experiment of acute toxicity of sliced Radix Aconiti Coreani and in compliance with the quality standard stipulated in pharmacopeia-surface features cross section colour and odor of sliced Radix Aconiti Co-reani the technology of processing Radix Aconiti Coreani has been optimized to be steaming the drug for four hours. PMID- 11038926 TI - [Comparison of immunity improving effects of different processed cortex Eucommiae]. AB - The immunity improving effects of different processed Cortex Eucomiae was compared. The results show that the effect of post-processed drugs is obviously stronger than that of pre-processed ones. No obvious difference have been found in the action of the post-processed drugs. PMID- 11038928 TI - [Content determination of baicalin and baicalein in shuanghuanglian powdery injection by the method of HPLC-ECD]. AB - The contents of baicalin and baicalein in Shuanghuanglian Powdery Injection have been determined by the method of HPLC-ECD under following conditions: column-ERC ODS-1161, 6 mm x 100 mm; temperature-55 degrees C; mobile phase-0.1 mol/L KH2PO4, acetic acid and tetrahydrofuran (1000:100:190); flow rate-0.8 ml/min; detector voltage-+650 mV. The result shows the limit of detection for baicalin and baicalein to be 25 ng and 50 ng respectively, with an average recovery of 100.1% and 99.7% respectively. Highly reproducible, rapid, simple, convenient and accurate, this method is good for the content determination of baicalin and baicalein in Chinese medicinal herbs and preparations. PMID- 11038927 TI - [Determination of mangiferin in qingqiliangying injection by reversed-phase HPLC]. AB - A reversed-phase HPLC method has been established for the determination of mangiferin in Qingqiliangying Injection. The mangiferin was separated on a Nova pak C18 column (150 mm x 3.9 mm) and detected at 258 nm, using methanol tetrahydrofuran-0.85% H3PO4 aqueous solution (500:80:15) as the mobile phase. The average recovery of mangiferin was 100.9%. The method is simple, rapid, and well reproducible, and thus very reliable for the quality control of Qingqiliangying Injection. PMID- 11038929 TI - [Structure of rubescensin E]. AB - Rubescensin E(I), a new diterpenoid, was isolated from the leaves of Rabdosia rubescens collcted in Jiyuan county, Henan Province On the basis of spectroscopic evidences, the structure of rubescensin E lished as ent-7 beta, 20-epoxy-7 alpha, 14 alpha-dihydroxy-6 alpha, 15 alpha-diacetoxy-16kaurene. PMID- 11038930 TI - [Phenylethanoid glucosides from flos Buddlejae]. AB - Four phenylethanoid glucosides were isolated from the flower of Buddleja officinalis. On the basis of specteral data, they were identified as salidroside(1), verbascoside(2), isoverbascoside(3) and echinacoside(4). Compounds 1, 3 and 4 were obtained from the plant for the first time. Compound 2 showed antibacterial and anticancer activities. PMID- 11038931 TI - [High performance liquid chromatographic determination of quercetin in the extract of Ginkgo biloba L. leaves]. AB - This paper reports the HPLC determination of one of the flavones quercetin in the extract of Ginkgo biloba leaves. Quercetin was separated completely on an YWG-C18 column(0.46 cm x 15 cm), using a mixture of methanol, water and phosphoric acid(50:49.8:0.2) as mobile phase, with UV detection at 360 nm. The results have shown the recoveries to be between 95.3%-103.2% and RSD 1.86%. PMID- 11038933 TI - [Laboratory study on pharmacology of Chinese medication qizhi weitong powder]. AB - The Chinese medication Qizhi Weitong Powder has been proved very helpful in depressing gastric evacuation and small intestinal propel reducing gastric secretion, lowering acidity of gastric liquid and activity of pepsin as well as enhancing peristalsis of domestic rabbits. Other effects such as distinct protection to ligated ulcer of pylorus and painkilling have also been confirmed. PMID- 11038932 TI - [A primary study on antagonizing effects of anti-snake venom Chinese herbs on endothelin-1 and sarafotoxin 6b]. AB - Some anti-snake venom Chinese herbs were used to test the antagonizing effect on ET-1 and S6b. Oral administration has shown that both the water and alcohol extracts from Cissus assamica and Aristolochia fordiana, and the water extracts from Desmodium microphyllum, Cynanthum paniculatum and Polygonum cillinerve are very helpful in reducing the acute death caused by ET-1 and S6b(i.v.), while the extracts from Cissus assamica, Aristolochia fordiana and Cynanthum paniculatum can dilate the vasoconstriction by ET-1 in a dose dependent manner. Pharmacodynamic parameters have shown that the potencies of alcohol extracts from Cissus assamica and Aristolochia fordiana are greater than that of Cynanthum paniculatum. These data suggest that traditional anti-snake venom herbs have antagonizing effects on ET-1. PMID- 11038934 TI - [An experimental study on the effects of lingzhi spore on the immune function and 60Co radioresistance in mice]. AB - Lingzhi spore (2 g.kg-1.d-1, 10 times) was orally administered to mice, and studies were carried out on the carbon clearance, delayed-type skin hypersensitivity (DTH), antibody hemagglutinin value and spleen weight. The results showed that Lingzhi spore could increase the immune function, inhibit the reduction of white blood cells and raise the survival rate of mice after they were radiated by 870 rad 60Co. PMID- 11038935 TI - [Clinical and experimental studies of zhilibao oral liquor in treating mental retardation]. AB - In 110 cases of treating mental retardation with Zhilibao Oral Liquor, the overall effective rate is 74%. The results of experimental studies indicate that the drug can improve the experimental impairment of learning gain and memory retention, protect from acute cerebral ischmia, and alleviate the signs of yang deficiency in mice. PMID- 11038936 TI - [Advances in the chemical study of Epimedium]. PMID- 11038937 TI - [Ethnopharmacology of Phyllanthus emblica L]. AB - This paper deals with the ethnopharmacology of P. emblica, a traditional herbal medicine used by many peoples in the world. The paper introduces the biological characters, geographical distribution-patterns, chemical constitution and pharmacology of the plant. By cross-cultural comparative study, this paper indicates that there are 17 countries and nations of the world using various parts of P. emblica in their medical treatment. The medicinal plant is good for anti-hepatitis, anti-cancer, anti-tumor and regulation of stomachal function. The plant is also regarded as a traditional immunomodulator and a natural adaptogen. The result of the study reveals that P. emblica is an important traditional medicine with broad prospects. PMID- 11038938 TI - [A comparative study on the chemical components of Salvia miltiorrhiza Bge. collected from Zhongjiang of Sichuan and from other habitats]. AB - In this paper, the chemical components in the root of Salvia miltiorrhiza from three habitats (Zhongjiang of Sichuan, Henan and Shandong) were comparatively studied. The results show that the TLC chromatogram and contents of tanshinone IIA in the crude drugs are about the same, the sodium content appears to be lower in the sample from Sichuan, but the contents of other metallic elements are about the same in the samples from different habitats. PMID- 11038939 TI - [Growth development of Ligustium chuanxiong Hort]. AB - Experimental studies and chemical analysis have been made on Ligustium chuanxiong in its period of growth, generation and growth of organs, content and accumulation of nutritive elements, accumulation of dry matters, fertilizer consumption, etc. PMID- 11038940 TI - [Peroxidase assay over the course of dormancy relieving with low temperature in Fritillaria thunbergii Miq. by electrophoresis]. AB - In order to study the physiological basis of dormancy relieving with low temperature in Fritillaria thunbergii, isoperoxidase changes of bud, adaxial epiderm and inner tissues in scale were assayed by means of electrophoresis over the course of dormancy relieving. All the patterns in bud and different parts of scale change at different stages, and the peroxidase activity in bud and adaxial epiderm is higher than that in the inner tissue of scale. These results indicate both bud and scale join in the dormancy relieving. PMID- 11038941 TI - [Analysis of chromosome karyotype of Lycium chinense Mill]. AB - The chromosome karyotypes of wild Lycium chinese and its cultivated breeds for vegetable have been studied. The Chromosome 12 of both has a satellite. The former, with 24 chromosomes and 1B karyotype, is a diploid and its karyotype formula is 2n = 2x = 24 = 18m + 6sm. The chromosome number of the latter is 48. As a tetrploid, it has 2B karyotype and a karyotype formula 2n = 4x = 48 = 36m + 12sm. PMID- 11038942 TI - [Comparative study on nutmeg, mace and their processed products]. AB - The contents of volatile oil, myristicin, safrol and methyleugenol, as well as the composition and content of fatty oil in nutmeg, mace and their processed products were comparatively studied. The composition of volatile oil and the detection of trimyristin were compared by TLC. A scientific basis for the development and application of mace has thus been provided. PMID- 11038943 TI - [Influence of sulphur smoking on the content of coumarins in the root of Angelica dahurica (Fisch. ex Hoffm.) Benth. et Hook.f.var. formosana (Boiss) Shan et Yuan]. AB - The content of coumarins in the root of Angelica dahurica (Ad) treated in different time spans with smoke of sulphur was determined by HPLC. The result showed that in nine hours before Ad was smoked thoroughly the content of coumarins reduced rapidly with the increase of time, and there after, the reduction was not obvious. After being treated for nine hours, the total coumarins were reduced by 75.3% and after forty hours the ratio reached 77.9%. PMID- 11038944 TI - [Adsorption of flavonoids in Ginkgo biloba L. leaves by macroporous adsorptive resins]. AB - This paper compares the adsorptive properties and adsorption kinetics of ten kinds of macroporous adsorptive resins for flavonoids. The structures and properties of these resins were studied. The results showed that the AB-8 resin was a fine adsorbent. PMID- 11038945 TI - [Transformation of the mode of thinking and semi-bionic extraction]. AB - On the basis of exploratory exposition of semi-bionic extraction, the method is further discussed in the light of mode of thinking. PMID- 11038946 TI - [Chemical constituents of Cuscuta austrolis R.Br]. AB - Australiside A(1), a new diterpenoid glucoside, was isolated together with thymidine(2), caffeic acid(3), p-coumaric acid(4) and caffeic-beta-D-glucoside(5) from Cuscuta australis. Based on spectral analysis and physico-chemical properties the structures of the above-cited five compounds were identified. The structure of compound 1 was deduced as 4-oic acid-7-oxo-kaurene-6 alpha-O-beta-D glucoside. Compounds 2, 3, 4 and 5 were obtained from Cuscuta for the first time. PMID- 11038948 TI - [Constituents from the fruiting body of Ganoderma lucidum (Fr.) Karst]. AB - One compounds were isolated from the fruiting body of Ganoderma lucidum. Basis on chemical evidences and spectral analysis (MS, UV, IR, 1HNMR, 13CNMR), its structures were deduced as ergosta-7,22 E-dien-3-one. PMID- 11038947 TI - [Minor alkaloids from the capsule of Papaver nudicaule L]. AB - Two minor alkaloids were isolated from the capsule of Papaver nudicaule. On the basis of physicochemical methods and spectroscopic analysis, they were identified as chelidonine, 5-hydroxy-2-hydroxymethylpyridine. Chelidonine was isolated from this plant and 5-hydroxy-2-hydroxymethylpyridine was isolated from genus for the first time. PMID- 11038949 TI - [Determination of fumaric acid in Sarcandra glabra (Thunb.) Nakai by HPLC]. AB - A method for the determination of fumaric acid in Sarcandra glabra is reported. After the sample was pretreated with an anion exchange resin column, fumaric acid was determined on a Spherisorb C18 column, with methanol-0.02 mol/L monobasic potassium (20:80, pH2.5) as the mobile phase and detected at 210 nm. The method is simple and accurate. PMID- 11038950 TI - [Determination of arbutin in the herbs of Vaccinium vitis-idaea L. by RP-HPLC]. AB - Determination of arbutin in the herbs of Vaccinium vitis-idaea has been carried out by RP-HPLC, using Inertsil-ODS column (4.6 mm x 250 mm) and mobile phase of methanol and water (15:85), and detecting at 280 nm wavelength. The average content of arbutin is 4.44%, RSD = 2.93%; the recovery rate is 100.7%, RSD = 2.85%. PMID- 11038951 TI - [Senility-preventing effect of erlingcankang decoction]. AB - Experiments with Erlingcankang Decoction showed that when given to silk worms it could noticeably prolong the growth period of larvae and raise an average of 10 day survival of the male moths; when given to mice it could prolong their life; and when given to on old rats it could markedly raise the contents of SOD in the liver and red cells, lower the content of MAO-B in the brain, LF in the brain and adrenal gland and also LPO in the liver. PMID- 11038952 TI - [Inhibition of hydrogen peroxide production on chondrocytes induced by fulvic acid by ginger volatile oil]. AB - In order to investigate the effect of ginger on Kashin-Beck disease (KBD), the ginger volatile oil was taken as a scavenger and proved effective in inhibiting the production of hydrogen peroxide in chondrocytes induced by fulvic acid from KBD area. PMID- 11038953 TI - [Effect of shesheng gantai capsules on the inducement of interferon in mice and rats]. AB - Based on the cytopathic effect method, Shesheng Gantai Capsules have been proved able to induce interferon in mice and rats. The induced interferon accords with Type II interferon in nature. PMID- 11038954 TI - [Identification of Scutellaria barbata D. Don and its adultrants]. AB - Studies have been made on the comparison and identification of morphological and anatomical characteristics of six species Scutellaria barbata and its adultrants. Several tables and drawings are given. This paper may be helpful to pharmacognosists for identifying commercial crude drugs. PMID- 11038955 TI - [Observation of pollen morphology of medicinal Epimedium plants in Guizhou province by SEM]. AB - The pollens of 11 species and one variety of Epimedium grown in Guizhou Province have been observed by SEM. The morphological characters (especially ektexine sculpture) of the pollens are described and the correlation among pollen morphology plant forms and chemical constituents is discussd. PMID- 11038956 TI - [Physiological and biochemical changes of seeds of Coptis chinensis Franch. in stratification and the effect of ABA treatment]. AB - The seeds of Coptis chinensis need stratification to break dormancy. In this paper the changes of enzyme activities DNA contents and protein contents in stratification under refrigeration and outdoor temperature conditions, as well as the influence of ABA treatment were studied. PMID- 11038957 TI - [The plant regeneration of Salvia miltiorrhiza Bge. transformed by Agrobacterium]. AB - The hairy roots and crown galls of Salvia miltiorrhiza were obtained by infecting plant with A. rhizogenes (strain 15834, LBA 9402) and A. tumefaciens (strain C58). The transformed plants were regenerated light and transplanted form cultural medium into soil successfully. The plants transformed by A. rhizogenes have characteristics of short stems and develop hairy roots, those and trans formed by A. tumefaciens grow vigorously featuring, longer stems and well developed roots. Both biomass production and tanshenone content are higher than the original plant. PMID- 11038958 TI - [Electrophoresis assay on isoperoxidase in the course of seed dormancy relieving for Fritillaria thunbergii Miq. at two different low temperatures]. AB - The relieving of seed dormancy for Fritillaria thunbergii required 50 days at 8 10 degrees C and 80 days at 3-5 degrees C. During the relieving process, isoperoxidase pattern changes were studied by means of electrophoresis. The results showed that some new bands were detected, some original bands enhanced as the seeds were treated with a low temperature, and the changes of lsoperoxidase patterns were similar at two different low temperatures. PMID- 11038959 TI - [Influence of processing on the content of hypaconitine in the roots of Aconitnum coreanum (Levl.) Rapaics]. AB - In this paper, the contents of hypaconitine in the roots of Aconitum coreanum and its processed products were determined by high performance liquid chromatography. The result provides scientific basis for processing the roots of Aconitum coreanum. PMID- 11038961 TI - [Microscopic quantitative analysis of Rhizoma Coptis in Xianglian pill]. AB - This paper reports the standards for microscopic quantitative analysis of Rhizoma Coptis. By comparing the microscopic quantitative analysis with spectrophotometry, the content of Rhizoma Coptis in Xianglian Pills has been determined and the accuracy of this method confirmed. PMID- 11038960 TI - [Determination of oleanolic acid contents in differently-processed Achyranthes bidentata Bl. by HPLC]. AB - The contents of oleanolic acid in differently-processed products of Achyranthes bidentata were determined by HPLC. The result showed the contents to be in the following sequence: salt-broiled product > alcohol steamed product > alcohol broiled product > unprocessed product. The average recovery rate was 100.2% and RSD 1.54%. The determination method has thus proved up to the standard for quantitative analysis. PMID- 11038962 TI - [Application of TLC to tannin measurement]. AB - There has been little report on the use of in measuring tannin in traditional Chinese medicinal preparations by TLC. Gallic acid, the decomposd product of tannin, was in bright yellow and green color, showing a strong specificity. The measurement of gallic acid can provide reliable indexes for quality assay of TCM preparations. PMID- 11038963 TI - [Separation and identification of the compounds from Achyranthes bidentata Bl]. AB - Eight compounds were separated from the roots of Achyrathes bidentata by repeated chromatography. On the basis of physicochemical properties and spectral analysis their structures were elucidated as alpha-spinasterol (1), beta-sitosterol (2), chrysophanol (3), dibutyl phthalate (4), palmitic acid (5), alpha-spinasterol-3-O beta-D-glucoside (6), daucosterol (7) and ecdysterone (8). Compounds 1-7 were isolated from the plant for the first time. PMID- 11038964 TI - [Chemical constituents of Begonia evansiana Andr]. AB - Six compounds were isolated from Begonia evansiana. By means of TLC, mp, 1H and 13CNMR, they were idenified as beta-sitosterol, beta-amyrin, daucosterol, stigmasterol, stigmasterol-3-O-beta-D-glucopyranoside and 4', 5', 7-trihydroxy flavone-6-O-beta-D-glucopyranoside. All the compounds were isolated from this plant for the first time. PMID- 11038965 TI - [Alkaloids in the seeds of Zyziphs jujuba Mill]. AB - Two alkaloids were isolated from the seeds of Zyziphs jujuba and identified on the basis of spectral data to be lysicamine and juzirine. These two alkaloids were obtained from the genus for the first time. PMID- 11038966 TI - [Chemical components from Rosa laevigata Michx]. AB - Laevigatanoside A (2 alpha, 3 beta, 19 alpha, 23-trihydroxy-12-ursorlic-28 glucopyester) was isolated from the fruits of Rosa laevigata for the first time. PMID- 11038967 TI - [Determination of flavonoids in the leaves of Hippophae L. by HPLC]. AB - A method for determination of three flavonoids in the leaves of Hippophae by high presure liquid chromatography was established. The contents of these flavonoids in the leaves of seven different species of Hippophae were determined and compared. The results provide a scientific basis for evaluating and utilizing the leaves of Hppophae. PMID- 11038968 TI - [The effect of glucoprotein component of musk on arachidonic acid metabolizing enzymes in rat polymorphonuclear leukocytes]. AB - To investigate the effects of musk-1, a glucoprotein component isolated from the water extract of musk, on arachidonic acid metabolizing enzymes, which include phospholipase A2(PLA2), 5-lipoxygenase (5-LO), and cyclooxygenase (COX), in rat polymorphonuclear leukocytes, an in vitro incubation system with rat polymorphonuclear leukocytes or homogenate supernatant of the same cells was used. In comparison with control, musk-1 at final concentrations of 1-100 micrograms/ml can increase AA release from PMNL by 6.0%-21.6%, decrease LTB4 biosynthesis in homogenate supernatant of the cells by 9%-81%, but increase 6 keto-PGF1 alpha production by 18.2%-85.4%. PMID- 11038970 TI - [Pharmacological study on magnetite]. AB - Magnetite can markedly inhibit the rodent turn-around reaction induced by acetic acid, reduce the threshold dose of pentobarbital sodium and shorten rodent's incubation period of falling asleep. It has also the following effects; antagonizing metrazol which causes rodent convulsions, postponing the incubation period of being startled by Huisuling, cutting down the extent of rodent's foot swells caused by JCCJ, and diminishing bleeding time and congulating time. PMID- 11038969 TI - [Direct protection of injured primary cultured hepatocytes of rats treated with carbon tetrachloride by salvianic acid B]. AB - The incorporation of [3H]thymidine obviously tends to decrease in the primary cultured hepatocytes of rats when the cells are treated with carbon tetrachloride. Meanwhile the activity of ALT markedly increases in the medium. But these pathologic alterations are inhibited by Salvianic acid B(SA-B) and the effect of 10(-6) mol/L SA-B is the best. PMID- 11038971 TI - [Liver-protective activity of Aralia taibaiensis Z.Z. Wang et H.C. Zheng]. AB - Compared with oleanolic acid and sodium chloride physiological solution, the water decoction of and the total saponin in root and cortex of Aralia taibaiensis were testified to have significant protective activity in experimental acute liver injury in mice induced by CCl4. PMID- 11038972 TI - [Pharmacological study on the compatibility of cortex Cinnamomi with Halloysitum Rubrum]. AB - The decoction of Cortex Cinnamomi (CC, 1 g/kg p.o.) and Halloysitum Rubrum (HR, 3 g/kg p.o.) or the combination of the two drugs (4 g/kg p.o., CC 1 g/kg, HR 3 g/kg) could antagonize the diarrhea caused by p.o. water ex tract of Radix et Rhizoma Rhei in mice; and inhibit the platelet aggregation induced by ADP in vitro. Meanwhile, the effect of the combination of the two drugs was not different from that of each single one. In addition, CC was able to inhibit the spontaneous movement of intestine in situ and showed an analgesic effect (hot plate method) in mice; HR was ineffective in these aspects and did not reduce the effect of CC. CC(20 g/kg p.o., i.p. or i.v.) exhibited very strong toxicity in mice, while HR(60 g/kg p.o., i.p. or i.v.) was nontoxic. When the two drugs were used together, the toxicity was markedly reduced. PMID- 11038973 TI - [A review of the study on tu fu ling (Smilax glabra Roxb.)]. PMID- 11038974 TI - TCM stage differentiation treatment of diabetic gangrene--an observation on microcirculatory changes. AB - Microcirculatory changes and therapeutic effects in 39 non-insulin dependent diabetes mellitus (NIDDM) cases were evaluated. Under the same basic treatment in both groups, the treatment group (21 cases) was treated with TCM drugs according to its stages and compared with the control group (18 cases, treated with 654-2). The results showed that the microcirculatory cumulative scores were significantly decreased in both groups (P < 0.05), but the decrease in the treatment group was more marked (P < 0.05). In the treatment group, 15 cases were cured, 3 markedly effective, 2 effective and 1 ineffective; while in the control group, 13 cases were cured, 1 markedly effective and 4 ineffective. Statistically, there is no significant difference between the 2 groups (P > 0.05) in therapeutic effects. PMID- 11038975 TI - An enzymologic study on bone marrow cells in patients with aplastic anemia treated by supplementing the kidney and removing blood stasis. AB - The content of cytochrome oxidase (CCO), succinate dehydrogenase (SDH) and neutrophil alkaline phosphatase (NAP) in bone marrow cells in 68 cases of aplastic anemia before and after treatment was determined by computerized graphical analysis and compared with that of normal volunteers (control group). The significantly lowered CCO and SDH levels and the markedly increased NAP content before treatment (P < 0.01) became approximately normal after that of supplementing the kidney and removing blood stasis (P > 0.05). PMID- 11038976 TI - Treatment of incipient diabetic nephropathy by clearing away the stomach-heat, purging the heart fire, strengthening the spleen and tonifying the kidney. AB - It is the view of the authors that accumulation of heat and dampness in the heart and stomach and deficiency of both spleen and kidney with blood stasis would be the main predisposing factors in pathogenesis of incipient diabetic nephropathy. A controlled clinical study of 32 patients treated with modified recipe of Wang Kentang's Bai Fu Ling Wan, a proved formula of clearing away the stomach-heat, purging the heart fire, strengthening the stomach and tonifying the kidney, and simultaneously resolving blood stasis and turbid qi was performed. The total effective rate was 93.8%, superior to that in the control group treated with Capoton (59.9%). A marked improvement in symptoms, control of blood glucose level and reduction of urinary protein obtained in the treatment group suggests that this treatment is effective in retarding the development of the disease. PMID- 11038978 TI - TCM treatment of asthma in children with yin hua wu mei tang--Professor Liu Bichen's experience in treating asthma. PMID- 11038977 TI - TCM treatment of diabetic hearing loss--an audiological and rheological observation. AB - 35 cases (48 ears) of diabetic hearing loss were randomly assigned to 2 groups: the treatment group of 19 cases (25 ears) treated with drugs for nourishing the kidney-yin, invigorating blood circulation and removing blood stasis based on differentiation of symptoms and signs, and the control group of 16 cases (23 ears) treated with euglucon and other routine drugs. The results showed that the total effective rate for audition elevation in the treatment group was 52.0%, and that in the control group, 26.1%. In the treatment group, the rheological indexes were improved after treatment, the difference being very significant in comparison with those before treatment (P < 0.05 and P < 0.01); while in the control group, except the whole blood low shearing specific viscosity, all the other 4 indexes were not statistically significant before and after treatment (P > 0.05). PMID- 11038979 TI - The effect of gan fu le in interventional treatment of hepatocarcinoma. PMID- 11038980 TI - Treatment of dizziness by massotherapy and traditional Chinese drugs--a report of 110 cases. PMID- 11038981 TI - Treatment of genual osteoarthritis by massotherapy. PMID- 11038982 TI - Treatment of protrusion of lumbar intervertebral disc by pulling and turning manipulations. PMID- 11038983 TI - Two hundred and seventeen cases of winter diseases treated with acupoint stimulation in summer. AB - 217 cases of chronic bronchitis and asthma were clinically treated and analyzed for the effects of combining electric stimulation with topical application of drug on acupoints. The results suggested that the combined therapy was superior to unitary therapy (P < 0.05). It is indicated that the combined therapy has a good curative effect in both short- and long-terms. PMID- 11038984 TI - Clinical observations on 109 cases of vocal nodules treated with acupuncture and Chinese drugs. AB - The treatment of vocal nodules with the combined use of needling on Kaiyin Yihao Xue [symbol: see text] Kai Yin Point No. 1 for Voice-Regaining) and medicinal spray and tea made according to Xie Qiang's empirical prescription, obtained a total effective rate of 93.6%, obviously superior to that of the group treated with western medicine and the group simply treated by Chinese drugs (P < 0.01). This suggests that the combined use of acupuncture and Chinese drugs in the treatment of vocal nodules has better therapeutic effect and requires shorter course of treatment. PMID- 11038985 TI - Efficacy and effect of SI17 therapy on pancreatic polypeptide in vascular and tension-type headache. AB - BACKGROUND AND PURPOSE: Vascular and tension-type headache is most commonly encountered, and SI17 therapy has been tested to treat headache with good results. The efficacy of SI17 therapy for vascular and tension-type headache was compared and the effect of SI17 therapy on pancreatic polypeptide (PP) was studied. MATERIALS AND METHODS: 29 cases of vascular headache (20 cases in acute attack during the trial) and 27 cases of tension-type headache (19 cases in acute attack) were enrolled in the study. Plasma PP level before and 4th day after treatment was measured by radioimmunoassay. RESULTS: SI17 therapy is better for the treatment of vascular headache. Vascular headache with higher PP level and tension-type headache with normal PP level had good therapeutic results. CONCLUSION: The clinical efficacy is better for vascular headache with the increase of vagus tension and for tension-type headache with normal vagus tension. PMID- 11038986 TI - An experimental study of effect of zheng tai instant powder on grand mal epilepsy. AB - Zheng Tai Instant Powder [symbol: see text] is a complex prescription of traditional Chinese medicine indicated for grand mal epilepsy. Its effect on central nervous system, energy metabolism, neurotransmitters and hemorheology were studied in animal models. The results demonstrated that the effect of Zheng Tai Instant Powder is mild, long-lasting and effective in treatment and prevention of epilepsy without any side effects. PMID- 11038987 TI - Actions of NO and INOS on endotoxin induced rat acute lung injury and effect of rhubarb on them. AB - This study is to explore the actions of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) on endotoxin (lipopolysaccharide, LPS) induced rat acute lung injury (ALI) and effect of Rhubarb on them. LPS was injected into the sublingual vein of male Wistar rats to prepare ALI animal models. The rats were divided into 4 groups: LPS, control, Rhubarb, and dexamethasone. Macroscopic and histopathological examinations of the lung specimens were performed and the biological indexes of lung, including wet weight/dry weight, the rate of neutrophils and protein content in the pulmonary alveolar lavage fluid, pulmonary vascular permeability and pulmonary alveolar permeability were observed. In the mean time, the contents of serum NO and the activities of lung tissue homogenate iNOS were measured. The results showed that in the LPS group, the injury and celluar infiltration in the pulmonary stroma and alveoli were more prominent than that in the control group. Lung wet weight/dry weight, the rate of neutrophils, protein content, pulmonary alveolar permeability, pulmonary vascular permeability were significantly increased (P < 0.01); NO and iNOS were also markedly elevated (P < 0.01). In the groups of dexamethasone and Rhubarb, the histopathological changes were significantly milder, and all the above biological indexes of lung injury and the contents of NO and the activities of iNOS were correspondingly decreased (P < 0.05). The above data demonstrate that NO and iNOS play an important role in the onset of ALI; dexamethasone and Rhubarb interfering treatment can ameliorate lung injury and decrease the concentrations of NO and the activities of iNOS, showing that through inhibiting the levels of NO and the activities of iNOS, these 2 agents exert protective effect on ALI induced LPS. PMID- 11038988 TI - Advances in TCM treatment of primary hepatocarcinoma. PMID- 11038989 TI - Acupuncture treatment of common cold. PMID- 11038990 TI - Comparison of pharmacological treatment versus acupuncture treatment for migraine without aura--analysis of socio-medical parameters. AB - This study was carried out in 120 patients affected by migraine without aura, treated in 4 public health centers and randomly divided into acupuncture group (AG) and conventional drug therapy group (CDTG). The evaluation of clinical results was made 6 and 12 months after the beginning of treatment and was worked out as well according to socio-medical parameters. Acupuncture was applied to the following points: Touwei (ST 8), Xuanlu (GB 5), Fengchi (GB 20), Dazhui (GV 14), Lieque (LU 7), treated with the reducing method. In AG, the figure scoring the entity and frequency of migraine attacks drops from 9,823 before treatment to 1,990 6 months after and 1,590 12 months after; while in CDTG, it drops from 8,405 before treatment to 3,927 6 months after and 3,084 12 months after. In AG, the total absence from work amounted to 1,120 working days/year, with a total cost (private + social costs) of 186,677,000 Italian liras. In CDTG, the absence from work amounted to 1,404 working days/year, with a total cost of 266,614,000 Italian liras. If we consider that in Italy the patients affected by migraine without aura are around 800,000, and that acupuncture therapy is able to save 1,332,000 Italian liras on the total average cost supported for every single patient, the application of acupuncture in the treatment of migraine without aura would allow a saving of the health expenses in Italy of over 1,000 billion liras. PMID- 11038991 TI - A clinical investigation on zhi ling tang for treatment of senile dementia. AB - Zhi Ling Tang (ZLT [symbol: see text], a TCM prescription designed for replenishing essence, supplementing marrow, invigorating qi, warming yang, removing blood stasis and phlegm, tonifying the brain, and invigorating mental activity) was used in 32 cases of senile dementia (SD). After treatment, the levels of serum cholesterol (Tch), triglyceride (TG), and plasma lipid peroxides (LPO) were lowered; the content of high density lipoprotein (HDL) and the activity of superoxide dismutase (SOD) in RBC significantly elevated, the cerebral blood flow was increased; latent period of P300 and P3 waves was shortened while the amplitude of P3 elevated; and topographic electroencephalogram, revised Hasegawa dementia scales (HDS) scores (P < 0.05 0.01) and clinical symptoms were improved. The total effective rate was 81.3%, indicating that ZLT exhibits relatively good therapeutic effects for treating senile dementia. PMID- 11038992 TI - Intermediate and late rheumatoid arthritis treated by tonifying the kidney, resolving phlegm and removing blood stasis. AB - Eighty-seven cases of intermediate and late rheumatoid arthritis were treated with Instant Shu Guan Wen Jing Granules ([symbol: see text] Relaxing Joints by Warming Channels) and Instant Shu Guan Qing Luo Granules ([symbol: see text] Relaxing Joints by Removing Heat from the Lung Channel) to tonify the kidney, resolve phlegm and remove blood stasis, and compared with 41 cases treated with Instant Wang Bi Granules ([symbol: see text] Prescription for Arthralgia syndrome). The treatment produced a clinical cure rate of 54.0% and a total effective rate of 90.8% as in against 29.3% and 73.2% respectively in the control group. The difference was significant (P < 0.01). Improvement in main symptoms and laboratory findings in the treatment group was all more marked than that in the control group (P < 0.05 or P < 0.01), with no side effects observed. PMID- 11038993 TI - A clinical investigation on tong xin luo capsule in treatment of coronary heart disease with silent myocardial ischemia. AB - 36 cases of coronary heart disease with silent myocardial ischemia (SMI) were treated sequentially with Tong Xin Luo (TXL) capsule and isosorbide dinitrate in random order. The results showed that both drugs were effective in decreasing episodes of SMI and shortening the duration of asymptomatic myocardial ischemia. TXL, however, was found with a better action and was superior to isosorbide dinitrate. It was also found that TXL could improve diastolic function of the left ventricle as well. PMID- 11038994 TI - Clinical observation on treatment of non-parvicellular carcinoma of the lung with jin fu kang oral liquid. AB - Jin Fu Kang Oral Liquid ([symbol: see text]), made of traditional Chinese drugs for supplementing qi and nourishing yin, was developed according to the common symptoms in lung carcinoma with deficiency of both qi and yin. Of the 96 cases in the Jin Fu Kang group, 1 case got complete remission (CR) after treatment, 8 cases partial remission (PR), 52 cases no change (NC), PR + NC covering 63.5%. Of the 52 cases in the group of Jin Fu Kang plus chemotherapy, 11 cases got PR after treatment, 26 cases NC, PR + NC covering 71.2%. Of the 25 cases in the chemotherapy group, 4 cases got PR after treatment, 11 cases NC, PR + NC covering 60.0%. The results show that the therapeutic effectiveness in the Jin Fu Kang group and the group of Jin Fu Kang plus chemotherapy was better than that in the chemotherapy group. The one-year survival rate and the two-year survival rate after treatment in the Jin Fu Kang group were 67.3% and 67.3% respectively; 66.7% and 66.7% in the group of Jin Fu Kang plus chemotherapy; and 40.3% and 0.0% in the chemotherapy group. The improvement of clinical symptoms, increase of body weight and improvement of health situation (KPS marks) after treatment in both the Jin Fu Kang group and the group of Jin Fu Kang plus chemotherapy were better than that in the chemotherapy group. Some indicators of immunology and hemogram after treatment were greatly improved in the Jin Fu Kang group, worse in the chemotherapy group, but no obvious improvement in the group of Jin Fu Kang plus chemotherapy. PMID- 11038995 TI - TCM treatment of bronchial asthma. AB - Xuan Fei Ding Chuan Wan ([symbol: see text] pills for facilitating the flow of the lung-qi to relieve asthma) and Xiao Chuan Gu Ben Wan ([symbol: see text] pills for treating asthma by consolidating the origin) were used for treatment of bronchial asthma. The cure rate in the treated group (n = 600) was 81.33%, significantly higher (P < 0.01) than that in the control group (n = 164, with a cure rate of 9.76%). PMID- 11038996 TI - Qi-promoting and phlegm-resolving method for treatment of diabetic microvascular complications. AB - OBJECTIVE: To evaluate the effects of qi-promoting and phlegm-resolving approach in treatment of diabetic microvascular complications. METHODOLOGY: Clinical observation of cases given modified Wen Dan Tang ([symbol: see text] Gallbladder warming Decoction). RESULT: Favorable results obtained in cases of diabetic microvascular complications of the type of stagnancy of qi and phlegm (diabetic retinopathy, diabetic nephropathy and diabetic foot). CONCLUSION: Wen Dan Tang is effective for diabetic microvascular complications of the type of stagnancy of qi and phlegm. PMID- 11038998 TI - Acupuncture techniques handed down from Dr. Zheng Kuishan's family. PMID- 11038997 TI - Empirical prescriptions used in treatment of epigastralgia due to blood stasis of deficiency type. PMID- 11038999 TI - Acupuncture ameliorates AIDS symptoms in 36 cases. PMID- 11039000 TI - Prof. Sheng Canruo's experience in acupuncture treatment of throat diseases with yan si xue. AB - Yan Si Xue ([symbol: see text]) refer to four points in the throat summarized by Prof. Sheng Canruo in his long-year clinical experience based on the combination of TCM theory and the knowledge of modern medical anatomy. By taking "Yan Si Xue" as main points, and other differential adjunct points, Prof. Sheng has obtained satisfactory therapeutic results in treating various throat diseases such as hoarseness, paralysis of vocal cord, dysphonia after radiotherapy on throat tumor, vocal nodules, disorder of the glossopharyngeal nerve, hysteric aphasia, and acute or chronic laryngopharyngitis. PMID- 11039001 TI - Effect of acupuncture on serum magnesium level in treatment of migraine. PMID- 11039002 TI - One hundred cases of sciatica treated by surrounding puncture. PMID- 11039003 TI - Electroacupuncture for treatment of 12 cases of infantile peroneal nerve injury. PMID- 11039004 TI - Eighty cases of injury of the superior cluneal nerve treated by electroacupuncture. PMID- 11039005 TI - Auricular-plaster therapy plus acupuncture at zusanli for postoperative recovery of intestinal function. AB - In order to relieve the abdominal distension and other discomforts due to gastrointestinal dysfunction after abdominal operations, the patients were treated by auricular-plaster therapy plus acupuncture at Zusanli (ST 36). 12 (92.4%) of the 13 cases in the treatment group showed recovery of normal peristalsis within 72 hours after operations, while that in 13 cases of the control group was 46.1%, indicating that auricular-plaster therapy plus acupuncture at Zusanli (ST 36) may promote postoperative recovery of the intestinal function. PMID- 11039006 TI - Therapeutic effect of massotherapy for prospermia. PMID- 11039007 TI - Treatment of protrusion of the lumbar intervertebral disc by intravertebral blocking and massotherapy--a report of 62 cases. PMID- 11039008 TI - Treatment of unstable angina pectoris based on disease- and syndrome differentiation. PMID- 11039009 TI - Effect of batroxobin on expression of c-Jun in left temporal ischemic rats with spatial learning and memory disorder. AB - The effect of Batroxobin on expression of c-Jun in left temporal ischemic rats with spatial memory disorder was investigated by means of Morri's water maze and immunohistochemistry methods. The results showed that the mean reaction time and distance of temporal ischemic rats for searching a goal were significantly longer than those of sham-operated rats, and at the same time c-Jun expression of left temporal ischemic region was significantly increased. However, the mean reaction time and distance of Batroxobin-treated rats were shorter and they used normal strategies more often and earlier than those of ischemic rats. The number of c Jun immune reactive cells of Batroxobin-treated rats was also less than that of ischemic group. In conclusion, Batroxobin can improve spatial memory disorder in temporal ischemic rats, and the down-regulation of the expression of c-Jun is probably related to the neuroprotective mechanism. PMID- 11039010 TI - Advances in TCM research and treatment of gastropathies associated with helicobacter pylorum. PMID- 11039011 TI - Acupuncture treatment of enuresis. PMID- 11039012 TI - [Multiple types of cytokines secreted by a mouse fetal liver stromal cell line]. AB - A murine fetal liver stromal cell line, called MFLC, has been established and maintained in DME supplemented with 10%NCS for 2 years. Without exogenous stimulation, the MFLC cells spontaneously secreted several types of cytokines, including IL-6, GM-CSF and chemotactic factor. Of which, IL-6 and chemotactic factor were abundant, GM-CSF at low level. Biological activities of IL-7 and IL-3 were not detected in the MFLC cell's supernatants. MFLC-SN induced colony formation of murine bone marrow cells in a dose dependent fashion in which mixed granulocyte/macrophage/megakaryocyte colonies(CFU-GMM) and granulocyte/macrophage colonies(CFU-GM) were dominant. MFLC-SN also sustained colony formation of bone marrow cells taken from 5-Fu injected mice, suggesting that there was a biological activity similar to SCF in MFLC-SN. The cytokines secreted by MFLC might play an important role in T cell early development in fetal liver. These results will be useful for us to analyse the mechanism of T cell development in fetal liver and to study the cytokine network regulation. PMID- 11039013 TI - [Comparative study on the chimeric ability of embryonic stem cell lines HDC and MESPU-13]. AB - While producing transgenic mice by ES cell route, it is important to know whether ES cells used have a strong ability to produce chimeras or not. In this report, two methods were used for studying the chimeric ability of two ES cell lines (HDC and MESPU-13). One method was the blastocyst microinjection, that is, chimeras were produced by injecting 15 ES cells into the cavity of C57BL/6J blastocysts. Another was GPI electrophoresis for examinizing the chimerism of ES cells in internal tisssues and organs of chimeras. The results showed that the ability of producing chimeras of MESPU-13 cells was stronger than that of HDC cells. The reason was discussed in detail. PMID- 11039014 TI - [The role of hydroxyurea in globin gene expression and cell differentiation of human erythroleukemia cell line]. AB - The HEL cells, a human erythroleukemia cell line, express mainly the fetal globin gene and small amount of the embryonic (epsilon) globin gene, but not the adult (beta) globin gene. Hydroxyurea, a small organic compound, has been successfully used to treat sickle cell anemia and beta-thalessaemia. Our data demonstrated that the growth rate of HEL cell proliferation was inhibited by different doses of hydroxyurea (from 50 mumol/L to 200 mumol/L). Using both routine RT-PCR and quantitative PCR analyses, we revealed that the expression of beta-globin gene was sharply activated and alpha-globin gene was almost completely silenced when HEL cells were induced for 3 or 5 days. Meanwhile, gamma-globin gene was expressed with no much difference between induced and uninduced HEL cells. We also demonstrated that the expression of GATA-1 and NF-E 2, which were two of the most important transcription factors in erythrocyte development, was activated about 3 folds in the induced cells. We suggested that the induction of GATA-1 and NF-E 2 expression by hydroxyurea might lead to activation of the adult beta globin gene through some pathways of signal transduction, therefore, hydroxyurea might play a role to induce HEL cells to terminal differentiation. PMID- 11039015 TI - [Effect of retinoic acids on phosphatidylcholine specific phospholipase D in 7721 human hepatocarcinoma cell line]. AB - In order to elucidate the relation between cell signal transduction and cell differentiation, the effect of two differentiation inducers--all trans-retinoic acid (ATRA) and 13 cis-retinoic acid (13 cis-RA) on the specific activities (nmol/hr.mg protein) of phosphatidyl choline specific phospholipase D (PC-PLD) in 7721 human hepatocarcinoma cell line was studied. It was found that ATRA and 13 cis-RA increased the specific activities of membrane bound PC-PLD on 2nd and 4th day of cell culture respectively. If the PC-PLD activities were calculated as activity per bottle cells, the effect of ATRA on the 2nd day was also higher than that of 13 cis-RA, while the reverse was true on the 4th day, revealing a postponed effect of 13 cis-RA as compared with ATRA. The increase of PC-PLD was higher when the culture medium was refreshed every day than that when the medium was unrefreshed, suggesting that a factor might be existed in the fresh medium which could enhance the stimulating effect of retinoic acids on PC-PLD. The effect of ATRA on PC-PLD specific activity were higher on 2nd day than that on 4th day, owing to the accelerated proliferation of the cells on 4th day, leading to the increase of protein contents and decrease of the enzyme specific activity calculated on the base of protein contents. Whereas, the effect of 13 cis-RA on PC-PLD specific activity were higher on 4th day than that on 2nd day, because the increase of enzyme activity was higher than the increase of protein contents on 4th day. The relation between the elevation of PC-PLD and protein kinases was further studied and discovered that ATRA decreased both the specific activities of membrane bound and cytosolic protein kinase C (PKC) as well as tyrosine protein kinase (TPK) either in 2nd or 4th day of cell culture, which indicated that the increasing effect of ATRA on membrane bound PC-PLD was not resulted from the stimulating effect of PKC and TPK on PC-PLD, and its mechanism remains to be further investigated. PMID- 11039016 TI - [Effect of 6-dimethylaminopurine on the resumption of meiosis and parthenogenetic activation in mouse oocytes]. AB - Chromatin condensation and germinal vesicle breakdown (GVBD) in mouse cumulus free oocytes was blocked by 2 mmol/L of 6-DMAP in vitro. Mouse oocytes (M II) from superovulated female mice could be activated by 6-DMAP. The activation rates of oocytes (18-19 h post hCG) cultured in CZB medium with 2 mmol/L 6-DMAP for 0.5, 1, 2, 3 h, were 26.1%, 75.2%, 75.8%, 77.3% respectively (Table 1), and the cleavage rates of these activated oocytes were 88.2%, 73.2%, 67.0%, 58.4% respectively (Table 2). The parthenogenetic activation type induced by 6-DMAP was different from that produced by ethanol (Fig.1). PMID- 11039017 TI - [Expression of human 17 beta-hydroxysteroid dehydrogenase gene in BmNPV expression system]. AB - Estrogenic 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) plays a crucial role in the synthesis of estrogens, but it also produces negative action of estrogens in promoting the growth of hormone-sensitive cancers, especially in breast and prostate. The high specific activity can be taken as an important signal for the diagnosis of cancers. Recombinant rAcBm-NPV/17 beta-HSD virus which contains the human 17 beta-HSD cDNA under the control of polyhedron gene promoter is generated by cotransfection of the BmN cells with the transfer plasmid pVL/17 beta-HSD and wild BmNPV genomic DNA. 17 beta-HSD is maximally expressed 72 h and 120 h post infection in BmN cells and the 5th instar silkworm larvae respectively. At those time interval, intracellular and hemolymphic enzymatic activity reach 0.12 U/mg and 0.15 U/mg of protein which produced total activity of 0.97 U/1.5 x 10(6) cells and 4.7 U/larva. The expressed quantities in female larvae are a little higher than that in male larvae. The present data shows that Silkworm/BmNPV expression system can express 3-5 times higher than that of the richest human placenta. It also indicates that there is an apparent band with a molecular mass of 35 kDa using SDS-PAGE method, the size of which is similar to that of the crude enzyme from placenta. PMID- 11039018 TI - [Studies of a 35 KDa substance from human fetal liver on the regulation of hematopoiesis]. AB - About 30%-40% of hematopoietic stem cells in human fetal liver of 3-5 months are in S phase of cell cycle, much higher than the ratio of 10% of that in adult bone marrow. The existance of highly active hematopoietic stem cell proliferation stimulators is probably its molecular basis. CFU-S "suicide rate" in rats was adopted to detect the effective substance. Through several steps of separation, we obtained a relatively purified substance of 35 kDa, termed it as FLS-4. CD 34 positive cord blood cells were sorted and assayed for their response to FLS-4 in 3H-TdR incorporation assay. The response to FLS-4 alone was approximately 1 times the background response seen with no factor added. In combination with IL-6 and IL-3 produced response that was 2.9 and 6.5 fold respectively greater than that observed with no factor added, but was weakly in comparison with the effects of SCF. In combination with GM-CSF or IL-3, FLS-4 can stimulate the formation of blast-colonies. The results indicate that the FLS-4 is very likely to be a novel hematopoietic stem cell proliferation stimulator. In physical or biological characteristics, it exhibited a unique character different from IL-3, IL-6, GM CSF, SCF or FLT3 ligand those are known to have hematopoietic stem cell proliferation stimulating activity. During the period of active hematopoiesis in fetal liver, FLS-4 might be the candidate in triggering hematopoietic stem cells from resting G0 to S phase. PMID- 11039019 TI - [Isolation of the expression fragments with the probe pool of human chromosome 14 q 24.3 generated by microdissection]. AB - The strategy of isolating the band-specific expression fragments from the probe pool of human chromosome generated by microdissection was reported in present paper. A chromosome 14 q 24.3 band-specific single copy DNA library was constructed based on this probe pool. Using this pool DNA as probe to hybridize the human bone marrow cell cDNA library, 68 primary positive clones were selected from 5 x 10(5) cDNA clones. Of them 32 clones were got in second-round screening and designed as cFD 14-1-32. Finally, 24 bandspecific expression fragments were identified from these 32 positive clones by analysing the results of DNA hybridization. Those band-specific clones can hybridize to both 14 q 24.3 DNA and human genomic DNA, but have no hybridization signal with 17 q 11-12 DNA. Partial sequences of 13 fragments of them were sequenced and were identified as novel cDNA sequences as well as have some homology with known genes in NCBI database. Analysis of expression spectrum of cFD 14-1 suggested that the cDNA fragments thus obtained can be used to isolate the genes not yet be cloned in 14 q 24.3 region. PMID- 11039020 TI - [Inhibition of parvovirus H-1 on transplantable human hepatoma and its histological and histobiochemical studies]. AB - A transplantable human hepatoma model, the QGY-9204, was used in this study. The growth kinetics of hepatoma in nude mice were compared after injection of parvovirus H-1 into the tumor growth. Significant difference in growth curves were seen between injected groups with H-1 dosages of 5 x 10(7) PFU and 5 x 10(8) PFU and that of control. It indicated that parvovirus H-1 was capable of suppressing the growth of human hepatoma. Previous studies showed H-1 is oncotropic, oncosuppressive and oncolytic. For histological, ultrastructural and histochemical examinations, transplantable hepatomas were taken at different time interval post H-1 (1 x 10(8) PFU per tumor growth) injection. For H-1 DNA amplification and H-1 nonstructural protein expression, PCR and ABC approach in hepatoma paraffin sections were used. The H-1 treated groups exhibited obvious signs of necrosis. It started on 3rd day post infection (3 d.p.i.) and the area of necrosis enlarged consecutively on 7 d.p.i., 10 d.p.i. and 14 d.p.i., but none was seen in saline-injected group even on 14 d.p.i. H-1 virions were also detected in the damaged tumor cells with numerous vacuoles in cytoplasm. Specific band (908 bp) of H-1 DNA and ABC immunostaining indicated H-1 DNA replication and NS-1 expression in tumors of treated groups, their time course was well in accordance with that process of necrosis. These results suggest that parvovirus H 1 promotes tumor necrosis by its DNA replication and cytotoxic NS-1 protein expression, and thus, it inhibits hepatoma growth and induces oncosuppression and oncolysis. PMID- 11039021 TI - [Gene therapy using recombinant adenovirus carrying herpes simplex-thymidine kinase gene to treat mouse B 16 melanoma in vivo]. AB - Adenoviral transfer of the herpes simplex virus thymidine kinase (HSV-tk) gene followed by administration of gancyclovir (GCV) was used to treat B 16 melanoma of C 57 BL/6 mice. B 16 murine melanoma cells transduced in vitro by a recombinant replication-defective adenovirus containing the HSV-tk gene [Ad(HSV tk)] were highly sensitive to cell killing by GCV, and the IC50 was 0.1 microgram/ml. A significant "bystander effect" was observed when Ad(HSV-tk) infected and -uninfected B 16 cells were mixed. Direct tumoral injection of Ad (HSV-tk) into established B 16 melanoma in C 57 BL/6 mice and subsequent treatment for 6 d with GCV resulted in the growth regression and necrosis of tumor nodules, and the tumor size was approximately reduced to one-twenty fifth of that of control animals. Finally, the safety of this treatment approach was demonstrated by limited dissemination of virus using sensitive RT-PCR. HSV-tk mRNA was detected only in the tumor nodule. These data indicated that gene therapy using Ad(HSV-tk)/GCV may function as an effective and safe alternative for treatment of melanoma in vivo. PMID- 11039022 TI - [The diversity of human hematopoietic stem/progenitor cells: VIII. Sterological features of CD 34+ hematopoietic cells isolated from human bone marrow]. AB - Human CD 34+ hematopoietic cells, a distinctive cell population containing hematopoietic stem/progenitor cells, have the capability to highly self-renewal, differentiation into all lineages of committed progenitor cells and reconstitution of both long-term hematopoiesis and immune functions after transplantation. In our previous study, the morphological, ultrastructural and cytochemical features concerning CD 34+ hematopoietic cells obtained by a two step isolated systems of CIMS-100/FACS 440 with 100% purity were depicted. Based on these observations on CD 34+ hematopoietic cells under light microscope, SEM and TEM, we recently analyze the sterological features of CD 34+ hematopoietic cells with the aid of Quantimet 970 automatic image analyser so that the three dimension structures regarding CD 34+ hematopoietic cells could be further made clear. By a series of measures including image scan-->modulus transform-->shadow correction-->image store-->statistical analysis, some morphometric parameters of CD 34+ hematopoietic cells were obtained as the followings: diametrs 3.490-6.741 microns, perimeters 11.776-26.240 microns, surface area 9.565-35.686 microns2, form factors 1.048-1.840, nucleus-plasma ratio 0.55-0.72, mean light density 0.17675-0.65100, integral light density 2717.217-46661.000. These morphometric data combined with our previous results from morphology, ultrastructure and functional subsets of CD 34+ hematopoietic cells strongly demonstrate that CD 34+ hematopoietic cells are really a heterogenous population. The possible reasons causing heterogeneous are close associated with either different functional subsets or differentiation lineages. To our knowledge, this is the first identification of sterological features of CD 34+ hematopoietic cells. PMID- 11039023 TI - [Cloning of variable region genes of anti-tetanus toxoid antibody and their expression as three kinds of engineered antibodies in E. coli]. AB - The heavy and light chain variable region genes of anti-tetanus toxoid (TT) antibody and the heavy chain Fd genes were amplified and cloned through RT-PCR from mouse hybridoma cells. The sequences of VH and VK were determined. Fd gene fragments were expressed in E. coli. The ELISA results indicated that the expressed Fd showed antigen binding activity but was nonspecific. Furthermore, through SOE and PCR techniques, the VH and VK gene fragments together with ScFv linker were assembled into single chain antibody (ScFv) gene fragment. While together with human heavy chain CH 1 gene fragment and Fab linker, they were assembled into chimeric Fab gene fragment. The two assembled gene fragments were separately inserted into phagemid pHEN 1, which was a fd-based vector containing gene 3 encoding the minor coat protein. In presence of helper phage M 13-VCS the anti-TT phage-ScFv or phage-Fab were displayed on the surface of phage particles respectively. Results from phage-ELISA indicated that both phage antibodies were TT-specific. PMID- 11039024 TI - [Studies on meiotic delay and aneuploidies induced by colcemid (COM) in male mouse germ cells]. AB - Colcemid (COM) was tested for induction of meiotic delay and aneuploidies in meiotic metaphase II (MMII) of male(101/E 1 XC 3 H/E 1)F 1 mice post single intraperitoneum (i.p.) injection. The dose of 1 mg/kg of COM was used and sampled at 2, 6, 10, 14 and 18 h after COM treatment. The number of MMII and MMI were the highest at 2 and 6 h repectively, and decreased rapidly with lengthening of COM treatment and reached the lowest at 14 h; then increased. However, the ratio of MMII to MMI was always significantly higher than in solvent control at every sample interval. Under our experimental conditions, COM did not show aneugenicity in male mouse germ cells. The possible mechanisms by which COM caused meiotic delay and reasons that COM did not induce aneuploidy in MMII were discussed. PMID- 11039025 TI - [Modulation of T-cell-mediated immune responses by trichosanthin via antigen processing and presentation]. AB - It has been shown in our previous study that the lymphoproliferation to allo- or soluble antigens could be inhibited when T cells were cocultured with antigen presenting cells (APCs) pulsed with Trichosanthin (Tk), a plant protein purified from a Chinese medicinal herb Trichosanthes kirilowii Maximovich. In this paper, data are presented dealing with the mechanism by which the Tk functioned as a down modulator. APCs of human peripheral blood were first treated with one of inhibitors of antigen processing (chloroquine, leupeptin or cycloheximide), followed by pulsed with Tk, and then added into a T cell culture with stimulators PMA and A 23187. The suppression of lymphoproliferation was observed to be obviously diminished or totally disappeared. In contrast, when CyA was used to replace the Tk for pulse-treatment of APCs that had received the same pretreatment with the inhibitors, no significant change was detected for the suppression, suggesting that the Tk might use a different pathway to induce hyporeactivity and that the pathway was concerned in APC's function of antigen presentation. This view obtained support from our immuno-histochemical examination of the Tk-pulsed APCs. By preparing colloidal gold-labelled Tk (Tk-G particle), we were able to show that the Tk-G, when incubated with APCs and T cells at 37 degrees C for two hours and washed, was visualized under the electronic microscope to be bound to APCs', instead of T cells', membrane surface and internalized cells' into endosomes. And then the Tk-G particles were further identified within lysosomes. In this way, the Tk molecules were subjected to be processed as an external antigen and might be presented to T cells to activate certain T subsets that in turn mediated the immunosuppression. This course was capable of, as expected, being interrupted by antigen processing inhibitor, especially by chloroquine. PMID- 11039026 TI - [Effects of spinal cord extract on neurite growth of GABAergic and dynergic neurons of spinal cord in vitro]. AB - We use the crude extract from human fetal spinal cord to study the effect of endogenous substances of the spinal cord on neurite growth of E 12-15 mouse GABAergic and DNYergic neurons of the spinal cord in vitro. The results indicated that the crude extract had no effect on GABAergic neurons but had markedly neurite-promoting effect on DNYergic neurons when the protein concentration of the crude extract was 250 micrograms/ml. It suggests that there are endogenous substances in the crude extract of the human fetal spinal cord, which can promote neurite growth of spinal cord neurons and this effect is neuron type specific at particular embryonic age. PMID- 11039027 TI - [Nuclear reconstitution around purified E. coli DNA in cell-free extracts of Xenopus laevis eggs]. AB - In recent years we have succeeded in inducing various exogenous DNA to assembly typical nuclei in cell-free Xenopus egg extracts. However, it is not clear if prokaryotic chromosomal DNA can also form the nucleus in this eukaryotic system. In our experiment, we used the purified E. coli. chromosomal DNA to reconstitute the nucleus in the nuclear reconstitution extracts and found that typical nuclei can also be assembled around added DNA. We observed the process of nuclear reconstitution by using light microscopy and electronic microscopy. The scanning results of chromatin-like materials and assembled nuclei by microspectrophotometer showed that E. coli, chromosal DNA underwent obvious change from condensation to decondensation during nuclear reconstitution. Incorporation of alpha-32P-dCTP into DNA suggested that the reconstituted nuclei were capable of DNA replication. PMID- 11039028 TI - [Immunogold labeling electron microscopy showing vimentin filament anchored on nuclear pore complex]. AB - The relationship between intermediate filament and nucleus is an important question to be solved. By combining sequential cell fractionation with immunoblotting, we showed that the intermediate filament protein in turkey erythrocyte is vimentin. Using pre-embedment immunogold labeling techniques together with sequential cell fractionation, we showed that cytoplasmic intermediate filaments in turkey erythrocyte are labeled specifically by rabbit anti-vimentin antibody-protein A-gold. Moreover, we showed that the cytoplasmic filaments anchored on nuclear pore complex were also labeled by rabbit anti vimentin antibody-protein A-gold specifically. Our results demonstrated that cytoplasmic filaments anchored on nuclear pore complex was vimentin filament. The experiments indicated that vimentin filament may be anchored on nuclear pore complex by binding with Nup 180 and intermediate filament may be involved in nuclear transportation. PMID- 11039029 TI - [Changes of PKC isoforms in induced differentiation of HL-60 cells by ATRA and PMA]. AB - Human premyeloid leukemic cell line (HL-60) were differentiated along granulocytic or monocytic/phagocytic lineage respectively by the treatment of all trans retinoic acid (ATRA) or phorbol ester (PMA), which was demonstrated by morphological observation, NBT reduction test, specific and nonspecific assay of esterases. By use of immuno-histochemical method, the changes of protein kinase C (PKC) isoforms alpha, beta I and beta II after induced differentiation of HL-60 cells were studied. The results showed that ATRA led to 5.0, 2.8 and 4.2 times increase of PKC alpha, beta I and beta II isoforms respectively as compared with the undifferentiated control levels and translocation of PKC from membrane to cytosol. The level of PKC alpha and beta II isoforms in HL-60 cells induced by PMA were decreased, but the expression of PKC beta I was increased. Besides, PMA led these 3 kinds of PKC to be present in cell nucleus with different intensity. These results suggest that the sustained activation of PKC alpha and beta may be required for granulocytic differentiation of HL-60 cells, while nuclear translocation of PKC may relatively be an important step for differentiation of HL-60 cells to monocytes/macrophages. PMID- 11039030 TI - [Study on the organization of chromatin and chromosome in mouse spermatogenic cells by scanning electron microscopy]. AB - It remains unclear about the intermediate construction of chromosome due to its highly compact nature and the limitation in methods. The present study was designed to investigate the construction of chromatin and mitotic chromosome in situ with scanning electron microscopy. Mouse testes were selected as the material, because of in which the spermatogenic cells divide actively and successively to form the sperm. Such a feature would be able to study the structure of mammalian chromatin and chromosomes along with the change of nuclear cycle. The animal were perfused with 200 ml of 0.075 mol/L KCl hypotonic solution to remove blood and placed for 15-20 min on ice followed by 0.5% glutaraldehyde and 0.5% formaldehyde for fixing. Through treated by the routine process of fractured and freeze dried with t-butyl alcohol, the specimens were then coated with a 3 nm thick platinum and observed with Hitachi S-430 scanning electron microscopy. It was found that the hypotonic treatment with 0.075 mol/L KCl solution was suit for demonstrating the nuclear structure, when the organelles were well preserved. The chromatin fibers of 10-30 nm and 80-125 nm in diameter could be recognized in the interphase nuclei, which were arranged losely at the region of euchromatin, and folded with each other into chromatin masses at the region of heterochromatin, while the chromatin fibers with the diameter of 80-125 nm often could be viewed on the mitotic chromosomes. Since its presence in interphase nuclei and mitotic chromosomes, it was considered that the chromatin fibers with 80-125 nm in diameter might play a role in the condensation of chromosome, serve as a type of the intermediate structure. PMID- 11039031 TI - [Transacting factors binding to the positive control region (PCR) in the 5' flanking sequence of human beta-globin gene]. AB - Using gel mobility shift assay, three protein factors (Pa, Pb, Pc) binding to the positive control region (PCR) in the 5' flanking sequence of human beta-globin gene were detected in the nuclear extracts from mouse fetal liver at d 18 or d 19 of gestation. Competitive experiment showed that Pb and Pc could bind to GATA-1 motif, therefore could be the members of GATA family. While Pa was a new and developmental stage specific trans-acting factor, we suggested that the factor Pa was related to the activation of beta-globin gene. PMID- 11039033 TI - American College of Rheumatology 64th annual scientific meeting and Association of Rheumatology Health Professionals 35th annual scientific meeting. October 29 November 2, 2000. Philadelphia, Pennsylvania, USA. Abstracts. PMID- 11039032 TI - [Changes in membrane potential of cultured mouse neuron during mitosis]. AB - Primarily cultured dorsal root ganglion cells and olfactory bulb cells were dissected from 12 to 14-d-old fetal C57 BL/6 J mice. After the cells were cultured for about two weeks, the growing status of cells were observed and the membrane potentials(MP) were recorded. The results show that mean value of the MP in anaphase was -68 +/- 3.1 mV (SE, n = 3), same as in interphase: Beginning from telophase, the cell membrane in the equator contracted gradually to become a concave ditch, and the MP decreased obviously, the mean value was -23.3 +/- 3.3 mV (SE, n = 6). After this, MP recovered gradually, till it divided into two sister cells. MP which were recorded separately in two sister cells were similar. But usually MP did not recovered to their normal values immediately. PMID- 11039034 TI - The American Association for the Study of Liver Diseases (AASLD) 51st annual meeting and postgraduate courses. October 27-31, 2000, Dallas, Texas, USA. Abstracts. PMID- 11039035 TI - The activity of glucosyltransferase adsorbed onto saliva-coated hydroxyapatite. AB - This study aimed to determine physical and kinetic properties of glucosyltransferase (GTF) adsorbed onto hydroxyapatite (HA) surfaces. For development of a solid-phase enzyme assay, 4.0-mg samples of washed HA powder were exposed to centrifuged whole saliva (WSHA) or buffer, and subsequently exposed to a GTF solution. The activities of GTF adsorbed to HA and that remaining in solution were measured. WSHA was more effective in adsorbing GTF than was naked HA. Enzyme activity on the surface of WSHA was enhanced; more activity was detected on WSHA than was apparently removed from solution. A similar effect was observed when GTF was adsorbed to naked HA from a mixture with lysozyme or saliva; however, no enhancement was seen when GTF was adsorbed from a mixture with albumin. Compared with GTF in solution, adsorbed GTF displayed activity over a much wider range of pH values. Temperature-activity profiles indicated that GTF adsorbed to surfaces had a lower temperature optimum (40 degrees C) than did soluble enzyme (45 degrees C), and that the bound enzyme was more resistant to adverse effects of heat at elevated temperatures. The majority of glucan made by GTF adsorbed to parotid saliva-coated HA remained attached to the surface. The activity of lysozyme adsorbed to HA was reduced by adsorption of GTF to the same surface and was almost completely abolished by formation of glucans by the adsorbed GTF. These results suggest that soluble bacterial enzymes found in saliva can be incorporated into pellicle, interact with host-derived molecules on the surfaces of teeth, express enzymatic activity, and potentially influence the biological properties of pellicle. PMID- 11039036 TI - A preliminary report of long-term elimination of detectable mutans streptococci in man. AB - Application of an antimicrobial varnish to the teeth of 33 adult volunteers resulted in the elimination of detectable mutans streptococci from the saliva of 21 of them for a mean period of 34.6 weeks (range, 4 to 89 weeks) without additional treatment. The mean number of applications of varnish required for elimination was 3.14 (range, 1 to 5). Extensive examination of 10 subjects made free of mutans streptococci on the basis of saliva examination revealed no detectable mutans streptococci in their dental plaque. In 14 of the subjects in whom mutans streptococci were eliminated, they subsequently re-appeared after a mean period of 22.7 weeks (range, 4 to 71 weeks). Four out of the five recurrences that were treated were eliminated with only one additional varnish application. The treatment failed to provide long-term elimination of detectable mutans streptococci in 12 of the 33 treated subjects. No serious adverse reactions were observed in any of the treated subjects. The results indicate that it is possible to eliminate mutans streptococci from man in a safe and effective manner. PMID- 11039037 TI - Distribution of antigenic determinants between Actinomyces viscosus and Actinomyces naeslundii. AB - A total of 18 monoclonal antibodies was raised against whole cells of Actinomyces viscosus and Actinomyces naeslundii. The monoclonal antibodies were used to determine the cross-reacting patterns among 26 strains of these species. Eleven different antigenic determinants were found. The specificity profiles of the antibodies indicated that the antigenic determinants of A. viscosus and A. naeslundii were arranged in a complicated mosaic. Extensive cross-reactions occurred between A. viscosus strains and strains of "typical" and "atypical" A. naeslundii. However, cross-reactions were rare between the two groups of A. naeslundii. A. viscosus appears to occupy a "middle position" between the two A. naeslundii groups. In addition to their value in seroclassification, some of the monoclonal antibodies were found to be useful in the identification of these species. One monoclonal antibody appeared to be selective for the "typical" A. naeslundii group. A. viscosus and "atypical" A. naeslundii-specific antibodies were also found, though they did not label every strain in their respective clusters. A. viscosus detection might be improved if mixtures of monoclonal antibodies were used. PMID- 11039038 TI - The effect of desalivation on coronal and root surface caries in rats. AB - Although the presence of coronal caries is declining in much of the Western Hemisphere, the prevalence of root surface caries is likely to increase as teeth are retained longer than heretofore. At the same time, an increasing number of the population suffer from dry mouth as a result of taking prescription drugs, with an apparent concomitant increased susceptibility to root surface caries. This study attempted to develop an animal model which would aid in the exploration of the effects of desalivation and the development of root surface caries. Animals were desalivated, infected with Actinomyces viscosus and Streptococcus mutans (sobrinus) 6715, and fed a cariogenic diet. Coronal caries developed rapidly in the animals; sufficient disease was present after two weeks to permit evaluation of potential therapeutic agents. Alveolar bone loss and root surface lesions developed in three to four weeks. S. mutans (sobrinus) and A. viscosus established readily in all animals; however, as the investigation progressed, populations of the latter declined, possibly because of the highly acidogenic environment. This model will facilitate investigation of the influence of hyposalivation and help in the exploration of agents to alleviate the adverse effects of salivary gland dysfunction. PMID- 11039039 TI - Analysis of photo-initiators in visible-light-cured dental composite resins. AB - Seven commercial visible-light-cured (VL) dental composite resins were analytically studied for identification of the photo-initiator consisting of photo-sensitizer and reducing agent. Gas-liquid chromatography (GC) was used for the determination of the dilute components extracted from the composite resin. Mass spectroscopy (GC-MS) was used for confirmation of the qualitative data obtained by GC. The results showed that all composite resins examined included camphorquinone (CQ) as a photo-sensitizer. The concentration of CQ in the resin phase, however, ranged from 0.17 to 1.03% w/w. The composite resin with hybrid sized filler tended to have a higher concentration of CQ than did the micro filled composite resin. As for the reducing agent, two out of seven brands contained dimethylaminoethyl methacrylate (DMAEMA), and one included dimethyl-p toluidine (DMPTI). The mixing ratio between CQ and the amine in these three composite resins also varied. Another four brands did not contain either DMAEMA or DMPTI, and would utilize different reducing agents. PMID- 11039040 TI - Adhesive 4-META/MMA-TBB opaque resin with poly(methyl methacrylate)-coated titanium dioxide. AB - An adhesive opaque resin for veneering on cast metal was developed with 4 methacryloxyethyl trimellitate anhydride and poly (methyl methacrylate)-coated titanium dioxide prepared by aqueous phase polymerization. The opaque resin was a modified 4-META/MMA-TBB resin. The powder consisted of 20% of the encapsulated material and 80% PMMA instead of pure PMMA powder. This resin hides the metal color when the thickness of the resin is as thin as 50 microns. The opaque resin bonded strongly to both cobalt-chromium alloy and visible-light-cured veneering resin. This self-curing opaque resin is applicable not only for bonding veneering resin to an alloy surface but also for bonding fixed partial dentures to enamel surfaces. PMID- 11039041 TI - Reliability of visual analog and verbal descriptor scales for "objective" measurement of temporomandibular disorder pain. AB - Eight dentists viewed standardized videotapes showing palpations of the temporomandibular joint and muscles of mastication and recorded their judgments concerning the amount of pain the patient was experiencing. Judgments were recorded using a four-point verbal descriptor scale (VDS) ("none", "mild", "moderate", "severe" pain) or a 100-mm visual analog scale (VAS) anchored with the terms "no pain" and "worst pain possible". Test/re-test reliability over a one-week period and interjudge reliabilities were calculated for each scale; reliabilities of the two scales were directly compared based on the statistical equivalence of weighted kappa and the Intraclass Correlation Coefficient. Neither scale showed satisfactory reliability. Median test/re-test reliabilities were k = 0.590 for the VDS and r = 0.822 for the VAS. Interjudge reliabilities averaged k = 0.394 for the VDS and r = 0.735 for the VAS. Direct comparison of reliabilities for the two scales showed no clear advantage for either scale. The marginal reliabilities of these scales, when used by dentists to quantify the patient's pain, suggest that neither scale should be regarded as an "objective" pain measure. PMID- 11039043 TI - Comparison of two scoring methods for root surface caries in hamsters. AB - The aim of this investigation was to compare two methods of scoring root surface caries in hamster teeth. The experimental material was mandibles from hamsters which had been infected with Actinomyces viscosus, fed a caries-promoting diet, and received either 0, 1, 10, or 25 ppm fluoride in the drinking water during an 18-week experimental period (Stookey, 1986). Root surface caries on the lingual surfaces of mandibular first molars was scored according to the grid method of Doff et al. (1977) and by the quantitative planimetric method of Firestone et al. (1986). Data from both methods confirmed that 25 and 10 ppm fluoride were significantly more caries-inhibitory than 1 ppm fluoride, which, in turn, significantly reduced root surface caries compared with the 0 ppm control group. Correlation coefficients for caries scores (0.92) and ranking of the scores (0.88) for each animal between the two methods were highly significant (p < 0.0001). The planimetric measurement of root surface caries in hamsters was as sensitive as the grid-scoring method. PMID- 11039042 TI - Sialographic images of pathological changes in the mouse parotid gland. AB - In order to correlate pathological changes of mouse parotid glands with their sialographic images, we conducted studies on mice with known pathological changes: mice with Stensen's duct ligated, NZB mice with a systemic auto-immune disease, and aged mice. The sialographic images were found to be specific for the pathological changes: The glands after ligation of Stensen's duct were characterized by dilated, large excretory ducts with a reduced system of peripheral ducts; the glands of NZB mice showed lobular leakage of the contrast medium from small excretory ducts; and the glands of aged mice showed a great reduction in the ductal system. It is concluded that sialography yields useful information on the pathological changes of the ductal systems in the mouse parotid gland. PMID- 11039044 TI - The effect of long-term use of a dentifrice containing zinc citrate and a non ionic agent on the oral flora. AB - The effect on the oral ecology of daily use, for seven months, of a dentifrice containing 0.5% (w/w) zinc citrate and a non-ionic agent, i.e., 0.2% (w/w) Triclosan (2,4,4'-trichloro-2'-hydroxydiphenyl ether), has been monitored on 13 adult volunteers. Plaque and saliva were sampled monthly and the bacterial flora analyzed. Twenty-six volunteers used a placebo dentifrice as part of their normal oral hygiene for four months to establish the baseline microbial flora. The volunteers were then split into two equal groups: One group continued to use the placebo dentifrice; the other used the dentifrice containing zinc citrate and Triclosan. There was no evidence that seven months' use of a dentifrice containing 0.5% zinc citrate and 0.2% Triclosan caused shifts in the oral microbial ecology, nor was there any evidence of developing bacterial resistance to Triclosan. PMID- 11039045 TI - Gallotannins inhibit growth, water-insoluble glucan synthesis, and aggregation of mutans streptococci. AB - During screening for anti-plaque agents of plant origin, ethanolic extracts from Melaphis chinensis (Bell), the Chinese Nutgall, exhibited strong inhibition of glucosyltransferase (GTF), in vitro adherence and glucan-induced agglutination of Streptococcus mutans 3209 and S. sobrinus B13. More than 91% inhibition of water insoluble glucan synthesis from sucrose by GTF was noted at a concentration as low as 7.8 micrograms/mL. Bactericidal effects on other mutans streptococci, S. salivarius, and Actinomyces viscosus were also evident. Through chemical fractionation and analyses, along with bioassays, the active components were identified as gallotannins. PMID- 11039046 TI - The acoustical characteristics of the normal temporomandibular joint. AB - Both clinical and empirical evidence suggests that the "normal" temporomandibular joint produces noise during function. The purpose of this study was to determine the conditions under which these noises might arise. Joint sounds and mandibular movements were recorded simultaneously from 200 adults who had no previous history or present symptoms of TMJ pain or dysfunction. The joint sounds were recorded bilaterally by means of two separate miniature vibration transducers mounted on a common headband, and incisal point mandibular movements were measured by a magnetometer tracking system. Recordings were made while each subject opened and closed the mouth, first in a natural and comfortable manner, and then to maximum displacement. Two types of measurements were made: the appearance, onset, and duration of joint-propagated sounds in relation to relative mandibular position; and the spectral properties of the detected sounds. The results of this study showed that for natural opening and closing movements of the mandible, the "normal" TMJ was silent for all age groups. However, for the maximum displacement condition, detectable sounds appeared at the points of maximum displacement in both the opening and closing phases of the cycle in over 80 percent of the subjects. However, these sounds were, as a class, substantially longer than, and spectrally distinct from, abnormal joint sounds, suggesting that they arise from a distinct physical substrate. PMID- 11039047 TI - Influence of contraction mismatch and cooling rate on flexural failure of PFM systems. AB - The interactive influence of cooling rate and the sign and magnitude of thermal contraction difference between metals and ceramic veneers on bond strength have not been extensively analyzed, although numerous bond-test studies have been reported during the past two decades. A previous analytical study of residual incompatibility stress in bond-test specimens indicated that bond strength values may be of relatively little value if the residual stress state of the metal ceramic specimens is not considered. The objective of this study was to determine the influence of cooling rate and contraction mismatch on the flexural failure resistance of metal opaque-porcelain strips. Specimens were subjected to four point loading in an Instron testing machine until crack initiation occurred at the metal-ceramic interface. The residual stress states in the ceramic region were estimated from finite element stress analyses of the bond-test specimens by use of dilatometry data obtained at the cooling rate of 3 degrees C/min. The total stress induced from the residual stress and the applied flexural load was also determined for these specimens. Statistical analyses of the experimental data revealed that the slowly cooled specimens exhibited a significantly lower (p < 0.05) flexural strength compared with rapidly cooled specimens. Regardless of the cooling technique, metal-ceramic specimens with a negative thermal contraction difference (alpha m - alpha p < 0) failed at significantly lower (p < 0.05) flexural loads than did specimens with a positive thermal contraction difference. PMID- 11039048 TI - A comparative study of human periodontal ligament cells and gingival fibroblasts in vitro. AB - Both periodontal ligament and gingival tissue are thought to harbor cells with the ability to stimulate periodontal regeneration, i.e., formation of new bone, cementum, and connective tissue attachment. To understand further the role of these cells in the regenerative process, we compared human periodontal ligament cells and gingival fibroblasts, both derived from the same patient, same passage, in vitro. Protein and collagen production was significantly greater in periodontal ligament cells when compared with that of gingival fibroblasts. In addition, periodontal ligament cells had higher alkaline phosphatase levels when compared with those of gingival fibroblasts. PMID- 11039049 TI - Wear of composite resin restorations in primary versus permanent molar teeth. AB - The purpose of this investigation was to compare the wear rates of posterior composite resin restorations in primary versus permanent teeth. Based on theories of composite wear in primary molars, as well as on empirical observations, we hypothesized that wear would be greater in permanent tooth restorations. In Part I of the study, quantitative wear data from four different clinical trials (three permanent tooth studies and one primary tooth study) were compared. The same posterior composite restorative material was investigated in each of the four studies. In Part II of the study, quantitative wear data were drawn from a single clinical trial wherein the same experimental posterior composite material was placed in primary and permanent molars in the same children. In Part II, the baseline sample size included posterior composite restorations placed in 92 primary and 95 permanent molars in all children (ages 7-10). Quantitative wear data were obtained by the cast assessment method at baseline, six, 12, and 24 months. The 24-month sample size was based on 48 available primary molar restorations and 89 permanent molar restorations. The loss of primary restorations was due almost exclusively to natural exfoliations. Findings in Part I revealed no significant difference in the wear of primary versus permanent molar restorations. In Part II, wear findings for primary molar versus permanent teeth, respectively, were as follows (in micrometers): 47 versus 49 at six months; 86 versus 80 at 12 months; and 133 versus 131 at 24 months. With a Wilcoxon two-sample test and an alpha level of 0.05, there were no significant differences at any recall for the wear rate of primary versus permanent restorations. These results differ sharply from findings reported by others. PMID- 11039050 TI - Degradation of starch and its hydrolytic products by oral bacteria. AB - Selected strains of oral bacteria were analyzed for their ability to degrade wheat starch, maltose, maltotriose, and maltoheptaose. S. sanguis IUOM-11M and JC804, S. mutans 6715, S. salivarius IUOM-8, A. viscosus IUOM-62, and A. naeslundii ATCC 12104 degraded all four substrates. S. mutans NCTC 10449 degraded starch, maltose, and maltotriose, while A. viscosus ATCC 15987 degraded starch and maltose, and S. sanguis SS34 degraded only maltose. L. casei IUOM-14 did not degrade any of the substrates. Analysis of starch degradation products from S. sanguis IUOM-11M and A. viscosus IUOM-62 demonstrated oligosaccharides, maltose, and trace amounts of glucose for the former and oligosaccharides, maltotriose, and maltose for the latter. S. sanguis IUOM-11M alpha-glucosidase (EC 3.2.1.20) demonstrated a pH optimum of 6.5 and greatly enhanced activity from maltose cultured cells as compared with cells cultured in glucose or fructose. The presence of fructose in the growth medium prevented this enhancement of activity by maltose. Maltose inhibited sucrose-dependent synthesis of S. sanguis IUOM-11M insoluble polysaccharide and both primer-dependent and primer-independent synthesis of soluble polysaccharide. Maltoheptaose inhibited primer-dependent but not primer-independent soluble polysaccharide synthesis. Several oral bacteria have the ability to hydrolyze starch and to degrade further the products to acidogenic substrates. These products may also inhibit sucrose-dependent synthesis of polysaccharides, which enhances the production of the acidogenic substrate fructose. The results add further support to the growing body of evidence suggesting that caries-promoting properties of starch may be expressed only when starch is present in diets containing sucrose. PMID- 11039051 TI - The influence of aging on lysosomal acid DNase of the rat submandibular gland. AB - Lysosomal and cytoplasmic fractions were prepared from rat submandibular glands for investigation of the release of lysosomal acid DNase in relation to aging. It was found that the acid DNase activity ratio for cytoplasmic/lysosomal fractions in rats aged 27 months was higher than that in three-month-old rats. The release of acid DNase from the lysosomal fraction by shaking was markedly increased in the fraction from the older animals. PMID- 11039052 TI - The decline and fall of dentistry in The Netherlands. PMID- 11039053 TI - Analysis of the buffering systems in dental plaque. AB - A semi-micro method was used for investigation of the buffering properties of whole plaque, plaque fluid, and washed plaque bacteria. Artifacts associated with titration of samples containing live bacteria were noted and their effects estimated. All three sample types showed minimal buffering in the region of neutrality, with much stronger buffering in the regions pH 4-5.5 and pH 8-9. For the range pH 4-7, almost 90% of the total buffer capacity of plaque appeared to be accounted for by macromolecules of bacterial cell walls and plaque matrix. Extracellular buffers in plaque fluid removable by centrifugation contributed up to 11%. These buffers (probably soluble proteins, peptides, organic acids, and phosphate) are, potentially at least, capable of exchange with saliva. In vitro, bicarbonate (dissolved in the extracellular fluid) contributed only 2-5% of total buffering; there was no evidence of formation of carbamino compounds. However, in vivo, salivary bicarbonate may be important as a continually replenished source of additional buffering. PMID- 11039054 TI - Effects of inorganic orthophosphate and pyrophosphate on dissolution of calcium fluoride in water. AB - Calcium and fluoride release from excess solid calcium fluoride was monitored for 15-30 min in aqueous solutions containing various concentrations of inorganic orthophosphate and pyrophosphate. Low concentrations of these ions (1-10 mumol/L) considerably inhibited the rate of dissolution of calcium fluoride. This inhibition was pH-dependent, being reduced at pH values below 5. It is suggested that a reduced calcium fluoride dissolution rate, in the presence of phosphate, can account for the relatively slow loss of calcium fluoride from dental enamel observed in recent clinical studies. It also appears that calcium fluoride coated with phosphate may provide a pH-controlled slow release of fluoride that may be of clinical significance and a major component of the cariostatic mechanism of topically applied fluoride. PMID- 11039055 TI - A feedback method to determine the three-dimensional bite-force capabilities of the human masticatory system. AB - A feedback procedure is described that enables a subject to exert bite forces in certain specified directions during static contraction of the human jaw muscles. The output of a three-dimensional transducer is fed to a computer. The magnitude and direction of the resultant force are computed and visualized by a cross on the screen of the computer terminal. In a bite experiment, the subject is instructed to match this cross with a point on the screen, representing the desired bite-force direction. The procedure allows for determination of the range of possible bite-force directions and magnitudes for various locations on the dental arch and study of the concomitant recruitment patterns of the jaw muscles. Some examples of measurement are given. PMID- 11039056 TI - An autoradiographic study of periodontal development in the mouse. AB - Tritiated thymidine was injected into 10- and 13-day-old mice because at this age the third molar is at the appropriate stage of development. At set intervals, the mice were killed and the distribution of labeled cells within the dental papilla and follicle examined. The change in labeling index with time was measured for defined areas in the papilla and follicle. It was shown that, during the late bell stage of development, cells moved from the papilla into the follicle. It was concluded that the pulp, rather than the investing layer of the follicle, is the source of the periodontium and that growth of the pulp and periodontal tissues could generate an important force contributing to tooth eruption. PMID- 11039057 TI - Effect of NaF on cAMP accumulation, cAMP-dependent protein kinase activity in, and amylase secretion from, rat parotid gland cells. AB - Stimulation of amylase secretion from parotid glands by beta-adrenergic agonists is mediated by the activation of adenylate cyclase and the resultant increase in cellular cAMP. Since NaF is known to increase adenylate cyclase activity and cAMP accumulation in intact cells, we investigated whether it would stimulate amylase secretion from isolated rat parotid gland cells. The results provide evidence that the addition of NaF (0.01-10 mmol/L) increased cAMP concentration (1.5-2.8 fold) in, and amylase secretion (16-93%) from, isolated parotid gland acinar cells. NaF was found to increase cAMP-dependent protein kinase activity ratios (51-84%) in a concentration- and time-dependent manner. The data suggest that the stimulation of amylase secretion from parotid gland cells by NaF may be mediated by an increase in the cellular cAMP concentration, which exerts its effect, at least in part, by increasing the activity of cAMP-dependent protein kinase. PMID- 11039058 TI - Comparison of in vivo vs. in vitro bonding of composite resin to the dentin of canine teeth. AB - Dogs were utilized in a study to compare the bond strengths of dentin bonding agents made to dentin in vivo and then again in vitro in the same teeth 30 min, one day, one week, and one month post-extraction. No statistically significant differences were observed between bonds made in vivo and those made in vitro at any time period. Contamination of the dentin surfaces with blood or saliva lowered the bond strengths, but these could be restored to control values by re surfacing of the dentin with a bur. PMID- 11039059 TI - An in vivo study of microhardness and fluoride uptake in partially demineralized human enamel covered by plaque. AB - In this investigation, microhardness changes in partially demineralized human enamel were studied after in vivo use of fluoridated toothpaste systems. Flattened enamel specimens were demineralized in vitro and subsequently positioned in approximal positions in the prostheses of 27 participants for three weeks; the samples were plaque-covered in vivo. After brushing for one week with a non-fluoridated paste to achieve an in vivo equilibrium, participants brushed with an assigned product for a two-week period. This test was a six-way cross over design which used randomization of subjects. Five fluoridated and one non fluoridated paste were tested. Knoop hardness measurements were carried out on sound and on in vitro demineralized enamel, and after in vivo exposure of the enamel to the dentifrice treatments. The results showed that during the two weeks of in vivo exposure to the fluoride products, net rehardening of the demineralized enamel did not occur, and no correlation was observed between fluoride uptake into the demineralized enamel and changes in microhardness. That we failed to observe rehardening may be due to the fact that the duration of this study was too short for any net remineralization to have occurred, especially because the samples were constantly covered with plaque. Other possibilities, such as the type and/or severity of the lesions used in this study, may account for the lack of rehardening. PMID- 11039060 TI - The relationship between oxide adherence and porcelain-metal bonding. AB - The lack of a reliable bond test has hindered the elucidation of the mechanism for porcelain-metal bonding in dental systems, because a test capable of detecting differences among porcelain-metal bonds of various qualities is required before the reasons for these differences may be ascertained. A method was developed in the present study whereby specimens of alloys with differing physical properties may be deformed to a constant strain to yield a fracture surface suitable for measurement of the area fraction of retained porcelain by an x-ray spectrometric technique described previously. The method proved sufficiently discriminating that significant differences could be found in 48 of the possible 66 comparisons among alloys and treatments. Linear regression analysis revealed a strong correlation (r2 = 0.947) between the area fractions of retained porcelain measured in the present study and the oxide adherence strength values measured previously. This strong correlation, when considered in light of the literature evidence for the presence of an oxide layer at the porcelain-metal interface, provides compelling support for the oxide layer theory of porcelain metal bonding in dental alloy systems. PMID- 11039061 TI - In vitro bonding of prosthodontic adhesives to dental alloys. AB - In vitro tensile bond strengths were determined for three adhesive cements and two resin-bonded bridge cements to two alloys, each prepared by two methods: sandblasted Ni-Cr-Be alloy (I), electro-etched Ni-Cr-Be alloy (II), sandblasted Type IV gold alloy (III), and tin-plated Type IV gold alloy (IV). Storage conditions of 24 hours at 37 degrees C and 30 days at 70 degrees C were evaluated. The highest bond strengths were obtained for the electro-etched Ni-Cr Be alloy, and all bond failures were cohesive. At both 24 hours and 30 days, the adhesive cements had the highest bond strengths to the other alloy/surface preparations (I, III, and IV). The adhesive cements usually failed cohesively under these conditions, whereas the resin-bonded bridge cements failed adhesively at the cement-alloy interface. Storage for 30 days at 70 degrees C caused average decreases of 30%, 5%, 15%, and 32% for alloy/surface preparations I to IV, respectively. PMID- 11039062 TI - Fraunhofer diffraction of light by human enamel. AB - Fraunhofer diffraction patterns of human enamel samples were photographed with a helium-neon laser beam (lambda = 633 nm). The first-order diffraction angle was in reasonable agreement with a prediction based upon enamel prisms acting as a two-dimensional grating. These results support the hypothesis that enamel diffracts light because of the periodic structure of enamel prisms with interprismatic spaces, which act as slits. PMID- 11039063 TI - The in vitro uptake of fluoride by secretory and maturation stage bovine enamel. AB - The objectives of this study were to determine the specific surface area of secretory-stage and of maturation-stage enamel, to compare the fluoride uptake by isolated enamel at these two stages on a surface-area basis, and to examine the effect of the organic matrix on the fluoride uptake by whole enamel. Fetal bovine secretory and maturation stage enamel samples were collected, and a portion of the enamel at each developmental stage was treated with hydrazine for removal of the organic matrix. The specific surface areas of the enamel mineral, as determined by the multi-point BET method, were 59.3 m2/g in the secretory stage and 37.9 m2/g in the maturation stage. Whole and deproteinated enamel samples were equilibrated in buffered solutions containing 10(-5) to 10(-3) mol/L fluoride, and the uptake was measured with a fluoride specific electrode. The results indicate that the in vitro fluoride uptake was controlled solely by the surface area of the apatitic mineral and that the organic matrix did not contribute to the fluoride uptake. PMID- 11039064 TI - Intra-oral retention of fluoride by bovine enamel from amine fluoride toothpaste and 0.4% amine fluoride liquid application. AB - The purpose of this study was to compare in vivo fluoride accumulation in enamel and in enamel lesions from a single topical fluoride application and daily toothbrushing with a fluoride dentifrice. Amine fluoride preparations were used for both products. Intra-oral appliances with bovine enamel specimens were worn by volunteers during a period of seven weeks. After this period, the specimens were analyzed for fluoride uptake and change in demineralization susceptibility. The results demonstrated that lesions had a high fluoride uptake capacity. Fluoride content values increased by 25-30 micrograms/cm2 during a single topical application, as compared with 10-15 micrograms/cm2 during seven weeks of toothbrushing. About half the fluoride acquired as a result of topical treatment was lost during subsequent exposure to the oral fluids when no further fluoride supplementation was given. The uptake of fluoride by sound enamel was comparatively small, regardless of the use of fluoride dentifrice or application. The presence of mature plaque at the time of fluoride application did not affect the amounts of fluoride delivered. Acid susceptibility tests showed that the enamel solubility exhibited a negative correlation with fluoride content of the specimens. PMID- 11039065 TI - Tooth mortality in an adult rural population in Kenya. AB - This paper reports on the pattern of tooth loss in a random sample of 1131 adults aged from 15 to 65 years in a rural area of Kenya in which access to formal dental care is minimal. We found that the majority of the population retained most of their dentition in a functional state even up to the age of 65 years: In all age groups, more than 50% had at least 26 teeth present, and more than 90% had at least 16 teeth present. The prevalence of edentulousness was less than 0.3%. The principal cause of tooth loss in all age groups was caries, and this was true for all tooth-types except incisors, for which periodontal disease was the main cause of tooth loss. The cultural practice of removing lower central incisors was observed only in those over 40 years of age. More teeth were lost due to caries among women than among men, while the reverse was true for teeth lost due to periodontal diseases. In view of the fact that most people retain most of their teeth throughout life, it is suggested that the most appropriate strategies for dental health care in this population should be those promoting self care, rather than the introduction of a formal treatment-oriented approach provided by dentists. PMID- 11039066 TI - Taurodontism in 47,XXY males: an effect of the extra X chromosome on root development. AB - Effects of an extra X chromosome on root development were studied in males with a 47,XXY chromosome constitution. Occurrence of taurodontism in the permanent molars of the lower jaw was noted from orthopantomograms of 30 Finnish 47,XXY males, 16 of their first-degree relatives, and a sample of 157 normal males and females. Nine, or 30%, or the 47,XXY males had at least one mandibular molar which was classified as taurodont. Only hypotaurodont teeth were found, and the teeth affected were all either second or third molars. None of the control relatives showed taurodontism. In the population sample, four individuals, or 2.5%, had taurodont teeth. A change in the mitotic activity of the cells of the developing teeth is one possible factor that can affect root formation leading to the development of taurodontism. PMID- 11039067 TI - Effects on gingivitis of two different 0.4% SnF2 gels. AB - For nine months we monitored the periodontal health of 81 adolescents undergoing orthodontic treatment with fixed appliances, to determine whether daily use of a brush-on 0.4% SnF2 gel would be better than toothbrushing alone in maintaining periodontal health in these patients, and whether a gel supplying a high percentage of available Sn2+ ion would be more beneficial than a gel supplying a low percentage of available Sn2+ ion. The subjects were matched for age and sex and placed into a control group, which used toothbrushing alone, and two treatment groups, which used toothbrushing supplemented with daily use of a SnF2 gel. One treatment group used a gel with 98% available Sn2+ ion twice daily for the entire nine months. The other treatment group used a gel with less than 2% available Sn2+ once a day for six months, then twice a day for the remaining three months of the study. Clinical assessments (Plaque Index, Gingival Index, Bleeding Tendency, pocket depth, and coronal staining) were made before appliances were placed and at one, three, six, and nine months after appliances were placed. Results indicated that the group using the high-availability Sn2+ gel twice daily had significantly lower Gingival Index and Bleeding Tendency scores at the one-, three-, six-, and nine-month examinations than did the control group. The group using the low-availability Sn2+ gel showed no significant differences in these assessments from the control group. Neither treatment group showed significant differences from the control group in Plaque Index or pocket depth. In the group using the high-availability Sn2+ gel, one subject developed mild coronal staining, and two developed moderate staining. PMID- 11039068 TI - Abrasion biopsy in studies of mineral density of experimental enamel lesions. AB - The profile of mineral density of experimental enamel lesions was determined by means of sampling with a new abrasion biopsy technique. Three types of lesions were produced on bovine enamel slabs: (1) pre-softened in lactic acid buffer for 16 hr; (2) pre-softened and exposed to a test for acid resistance for seven days; and (3) pre-softened, exposed to the seven-day Intra-oral Cariogenicity Test with extra-oral immersions in 1000 ppm F solutions for one min twice daily (ICT/F), plus the seven-day test for acid resistance. Lesions were assessed with measurements of surface microhardness, microradiography of thin sections, and abrasion biopsy. For abrasion biopsy of the experimental lesions, 15 parallel layers of approximately 10 microns each were abraded simultaneously with reference slabs of sound enamel on strips of lapping film. The depth of abrasion for each sample was calculated from the phosphorus content of the reference sample. Exposure to the ICT/F formed an acid-resistant zone within the lesion which diminished the microhardness change, although it did not have an appreciable effect on the total lesion depth as assessed with microradiography; abrasion biopsy indicated the formation of a mineral-dense zone within the lesion. This recently-developed technique of abrasion biopsy of experimental lesions offers the opportunity to link the composition of the lesion to controlled experimental conditions which will improve our understanding of demineralizing and remineralizing reactions on a standard tooth substrate. PMID- 11039069 TI - Comments on the clinical application of fibronectin in dentistry. AB - Several studies have demonstrated that citric acid demineralization of the root surface promotes tissue attachment. Since demineralization exposes collagen to which fibronectin binds, the role of fibronectin in the attachment of cells to the tooth surface has been of considerable interest. It is clear that fibronectin and other cell adhesion proteins can promote cell attachment to the tooth surface; therefore, attempts have been made to utilize these findings in a clinical setting. Using a quantitative ELISA procedure to measure the binding of fibronectin to demineralized bone and tooth, we have found that 1 microgram fibronectin can saturate approximately 1 mg of either demineralized bone or demineralized tooth powder. Since serum contains 300 micrograms fibronectin per mL, the bleeding that occurs during oral surgery should saturate exposed tooth surfaces with amounts of fibronectin adequate for cell adhesion. Thus, exogenous fibronectin would appear to be of little clinical benefit. PMID- 11039070 TI - Preliminary observations on the effect of mantle field radiotherapy on salivary flow rates in patients with Hodgkin's disease. AB - Changes in stimulated and non-stimulated whole saliva flow rates were measured in 11 Hodgkin's disease patients who received therapeutic doses of radiation to a mantle field at the M. D. Anderson Hospital in Houston, Texas. Salivary flow rates were examined before, during, and after radiotherapy. Mean flow rate reductions of 54% for non-stimulated saliva and 55.7% for paraffin-stimulated saliva were observed post-radiotherapy. Flow rates had not returned to pre irradiation levels in any of the patients who were observed for two to three months after completion of therapy. Results obtained from this preliminary study indicate that most patients who receive therapeutic doses of radiation to a mantle field experience a significant reduction in salivary output which is manifest during the period of treatment and persists for a period of at least two to three months post-radiotherapy. PMID- 11039071 TI - Stimulating professional interaction among researchers and clinicians in dentistry. PMID- 11039072 TI - Disobeying the rules. PMID- 11039073 TI - Dental science and dental health: now and a century hence PMID- 11039075 TI - The American College of Surgeons 86th Annual Clinical Congress and the 55th annual sessions of the Owen H Wangensteen Surgical Forum. October 22-27, 2000. Chicago, Illinois, USA. Abstracts. PMID- 11039074 TI - On the dissolution of hydroxyapatite in acid solutions. PMID- 11039077 TI - Association of Peutz-Jeghers-like mucocutaneous pigmentation with breast and gynecologic carcinomas in women. AB - Most reports describe an increased risk of malignancy in Peutz-Jeghers syndrome (PJS). We identified individuals with PJS-like pigmentation but no polyposis, designated as isolated mucocutaneous melanotic pigmentation (IMMP), and 1) characterized their clinical features, 2) assessed them for cancer events, and 3) screened a sample of these subjects for mutations in LKB1, a gene responsible for a portion of PJS cases. Review of Mayo Clinic records from 1945 to 1996 identified 26 patients with IMMP. All were then interviewed or their medical records reviewed to determine if cancer had developed. Conformation-sensitive gel electrophoresis (CSGE) screening for LKB1 mutations was followed by direct sequencing. Ten of these 26 individuals (38%) developed 12 malignancies that arose in the cervix (n = 3), endometrium (n = 3), breast (n = 1), kidney (n = 1), lung (n = 2), colon (n = 1), and lymphatic tissue (n = 1). In females with IMMP, the relative risk for cancer was 3.2 (95% CI, 1.2-6.9), while that for males was not increased. The relative risk for breast and gynecologic cancers was 7.8 (95% CI, 2.5-18.1) in affected females. Of 9 individuals tested, no LKB1 mutations were detected. Classical PJS is associated with an increased cancer risk. Our results indicate that IMMP is another lentiginosis with cancer predisposition. In particular, the relative risk for cancer in females with IMMP was significantly increased, as is true in females with PJS. However, LKB1 mutations did not contribute to the development of IMMP in the patients tested. PMID- 11039076 TI - Visual manifestations of giant cell arteritis. Trends and clinical spectrum in 161 patients. AB - Giant cell (temporal) arteritis (GCA) is the most common systemic vasculitis in Western countries. It involves large and medium-sized vessels with predisposition to the cranial arteries in the elderly. Cranial ischemic complications, in particular permanent visual loss, constitute the most feared aspects of this vasculitis. Although the use of corticosteroids and a higher physician awareness may have contributed to a decrease in the frequency of severe ischemic complications, permanent visual loss is still present in 7%-14% of patients. To investigate further the incidence, trends, and clinical spectrum of visual manifestations in patients with GCA, we examined the features of patients with biopsy-proven GCA diagnosed at the single reference hospital for a defined population in northwestern Spain during an 18-year period. Predictive factors for the development of any visual manifestation, not only permanent visual loss, were also examined. Between 1981 and 1998, 161 patients were diagnosed with biopsy proven GCA. Visual ischemic complications were observed in 42 (26.1%), and irreversible blindness, mainly due to anterior ischemic optic neuropathy and frequently preceded by amaurosis fugax, was found in 24 (14.9%). Despite a progressive increase in the number of new cases diagnosed, there was not a significant change in the proportion of patients with visual manifestations during the study period (p = 0.37). Patients with visual ischemic complications had lower clinical and laboratory biologic markers of inflammation. Indeed, during the last years of the study, anemia was associated with a very low risk of visual complications. Also, HLA-DRB1*04-positive patients had visual manifestations more commonly. Patients with other ischemic complications developed irreversible blindness more frequently. The best predictors of any visual complication were HLA-DRB1*04 phenotype (odds ratio [OR] 7.47) and the absence of anemia at the time of admission (OR for patients with anemia = 0.07). The best predictors of irreversible blindness (permanent visual loss) were amaurosis fugax (OR 12.63) and cerebrovascular accidents (OR 26.51). The present study supports the claim that ocular ischemic complications are still frequent in biopsy-proven GCA patients from southern Europe. The presence of other ischemic complications constitutes an alarm for the development of irreversible blindness. In contrast, a higher inflammatory response may be a protective factor against the development of cranial ischemic events. PMID- 11039078 TI - Male pseudohermaphroditism due to 17 beta-hydroxysteroid dehydrogenase 3 deficiency. Diagnosis, psychological evaluation, and management. AB - Ten male pseudohermaphrodites with 17 beta-hydroxysteroid dehydrogenase 3 (17 beta-HSD3) deficiency were evaluated in 1 clinic with an average follow-up of 10.1 years. The diagnoses were made by demonstrating low to normal serum testosterone levels, high androstenedione levels, and high ratios of serum androstenedione to testosterone in the basal state or after treatment with human chorionic gonadotropin. The molecular features of the underlying mutations were identified in all 7 families. Two additional males in the same families are believed to be affected on the basis of history obtained from family members. All of the 46,XY individuals in these families were registered at birth and raised as females (despite the presence of ambiguous genitalia in all or most), and all virilized after the time of expected puberty due to a rise in serum testosterone to or toward the normal male range. The age at diagnosis varied from 4 to 37 years. Ten individuals were studied by the same psychologist, and change of gender role (social sex) from female to male occurred in 3 subjects and in the 2 presumed affected subjects not studied. The individual with the highest serum testosterone level maintained female sexual identity, and in 2 families some of the affected males changed gender role and others did not. Thus, while androgen action plays a role in the process, additional undefined psychological, social, and/or biologic factors must be determinants of gender identity/role behavior. Management of the 7 individuals who chose to maintain female sex roles included castration, clitoroplasty, vaginal enlargement procedures when appropriate, treatment of hirsutism, cricoid cartilage reduction, and estrogen replacement. Three of the 7 are married (2 twice), 1 is involved in a long-term heterosexual relationship, 1 is engaged to be married, and the other 2 are not married and not believed to be sexually active. The 3 subjects who changed gender role behavior to male underwent hypospadias repair, and 1 was given supplemental testosterone therapy. One of these men is divorced, and the other 2 (aged 29 and 35 years) are unmarried. The diagnosis in 8 of these subjects was made after the time of expected puberty; it is unclear whether the functional and social outcomes would have been different if the diagnosis had been made and therapy begun earlier in life. PMID- 11039079 TI - Hut lung. A domestically acquired particulate lung disease. AB - We report an illustrative case of advanced "hut lung," or domestically acquired particulate lung disease (DAPLD), in a recently emigrated nonsmoking Bangladeshi woman with a history of 171 hour-years of exposure to biomass smoke. She presented with symptoms of chronic cough, dyspnea, and early parenchymal lung disease. High-resolution computed tomography (CT) of the chest demonstrated numerous 2- to 3-mm nodules, sparing the pleural surface. To our knowledge, this is the first such report of CT findings in the literature. Bronchoscopy yielded typical anthracotic plaques and diffuse anthracosis with interstitial inflammation on histopathologic examination of biopsy specimens. DAPLD is potentially the largest environmentally attributable disorder in the world, with an estimated 3 billion people at risk. Caused by the inhalation of particles liberated from the combustion of biomass fuel, DAPLD results in significant morbidity from infancy to adulthood. Clinically, DAPLD manifests a broad range of disorders from chronic bronchitis and dyspnea to advanced interstitial lung disease and malignancy. While a detailed environmental history is essential for making the diagnosis in most individuals, for patients with advanced DAPLD, invasive modalities such as bronchoscopy with transbronchial biopsy and examination of bronchoalveolar lavage fluid help differentiate it from other diseases. Recognition of this syndrome and removal of the patient from the environment is the only treatment. The development of well-controlled interventional trials and the commitment of sufficient resources to educate local populaces and develop alternative fuel sources, stove designs, and ventilation are essential toward reducing the magnitude of DAPLD. PMID- 11039080 TI - Long-term outcome of idiopathic pulmonary hemosiderosis in children. AB - We retrospectively analyzed the long-term outcome of idiopathic pulmonary hemosiderosis (IPH) in 15 children. IPH started at a mean age of 5 years, and the mean duration of follow-up was 17.2 years (range, 10-36 yr). Four patients developed immune disorders, 3 cases of rheumatoid polyarthritis or rheumatoid polyarthritis-like diseases and 1 case of celiac disease. Respiratory outcome showed that 3 patients had severe symptoms: 2 patients developed severe pulmonary fibrosis resulting in major chronic respiratory insufficiency, and 1 patient had severe asthma. Twelve patients (80%) had mild or no respiratory problems and were able to lead a normal life. According to chest X-ray and pulmonary function test data, 4 patients had normal chest X-ray and no evidence of restrictive syndrome, 6 patients had an interstitial pattern on chest X-ray and evidence of restrictive pattern, 1 patient had an interstitial pattern but normal lung function, and 1 patient had a normal chest X-ray but evidence of mixed obstructive and restrictive pattern. Our results show that long-term survival is possible in patients with IPH. Factors of poor prognosis seem to be the presence of antineutrophil cytoplasm antibodies (ANCA) or other autoantibodies. No other clinical or biological predictive factors for prolonged survival were found. PMID- 11039081 TI - Streptococcus pneumoniae endocarditis in adults. A multicenter study in France in the era of penicillin resistance (1991-1998). The Pneumococcal Endocarditis Study Group. AB - To better define the overall characteristics and risk factors for dying of adult pneumococcal endocarditis (PE) focusing on the echocardiographic diagnosis, the impact of surgery, and emergence of penicillin resistance, the medical and microbiologic charts of adult PE cases observed between 1991 and 1998 in university and general hospitals were reviewed through a nationwide retrospective study in France. Thirty cases of PE (22 men, 8 women; median age, 53 yr; range, 27-87 yr) were collected and validated. Twenty patients (66.7%) had no known predisposing cardiopathy; 4 had a bioprosthetic valve. The primary focus of infection was pneumonia in 10 (33.3%), and meningitis was noted in 12 (40.0%). Half the patients suffered from chronic alcoholism. Echocardiography detected vegetation(s) in 29 cases (96.7%), valvular perforation in 6 (20.0%), and/or valve ring abscess in 4 (13.3%). The most frequent complications were congestive heart failure (n = 19), large arterial emboli (n = 8), and focal abscesses (n = 7). Five strains were penicillin-resistant. Twenty (66.7%) patients underwent valve replacement, 12 of them during the first month. The overall mortality rate was 24.1%. According to a multivariate analysis, the risk factors independently associated with dying were age > or = 65 yr and septic shock, while cardiac surgery was protective (p < 0.01). In conclusion, PE is usually fulminant and causes severe valve damage and embolic complications; its short-term prognosis might be improved by early valve replacement. PMID- 11039082 TI - Application of a prediction rule to select which patients presenting with lymphadenopathy should undergo a lymph node biopsy. AB - We conducted the present study to develop a clinical prediction rule for discriminating which patients with peripheral lymphadenopathy require a lymph node biopsy. The clinical features of 315 patients with peripheral lymphadenopathy were analyzed to develop the prediction rule: 83 had diseases requiring a lymph node biopsy (Lymph Node Biopsy Group [BG]), while 232 had diseases that could be diagnosed without a lymph node biopsy (Non-Lymph Node Biopsy Group [NBG]). Among 23 examined clinical covariates, we identified 6 that independently predicted the need for lymph node biopsy and were graded as follows: 1) Age: x1 = 0, if < or = 40 years and 1, if > 40 years. 2) Tenderness in palpation: x2 = 0, if absent and 1, if present. 3) Size of the greatest lymph node: x3 = 0, if < 1.0 cm2, 1 if 1.0-3.99 cm2, 2 if 4.0-8.99 cm2, and 3 if > or = 9.0 cm2. 4) Generalized pruritus: x4 = 1, if present and 0, if not. 5) Supraclavicular lymphadenopathy: x5 = 1, if present and 0, if not. 6) Texture: x6 = 1, if nodes are hard and 0, if not. The prediction rule was then validated in a subsequent group of 160 patients (32 in the BG; 128 in the NBG). A score Z = 5x1 5x2 + 4x3 + 4x4 + 3x5 + 2x6 - 6 corresponded to every patient, according to the results of logistic regression analysis. If patients with Z > or = 1 were considered to need lymph node biopsy, the sensitivity of the prediction rule was 95.2% (95% confidence intervals [CI]: 88.1%-98.1%) and the specificity was 81.0% (95% CI: 75.4%-85.6%). Within the Validation Group of patients the prediction rule was at least equally effective. Sensitivity was 96.9% (95% CI: 83.9%-99.5%) and specificity was 91.4% (95% CI: 85.1%-95.2%). The described rule can be useful in the clinical evaluation of patients with peripheral lymphadenopathy. Further validation by other groups is required, and its cost-effectiveness has to be investigated. PMID- 11039083 TI - [Questionnaire on venous thromboembolism prevention by low molecular weight heparin in medical environment]. AB - OBJECTIVE: The aim of this prospective study was to observe the prescription of low-molecular weight heparin (LMWH) for the prophylaxis of venous thromboembolic disease in medical patients. METHODS: We included all the patients on LMWH prophylaxis in 5 medical departments of Sainte-Marguerite Hospital in Marseille. The study described the reasons for this prophylaxis, the thrombotic risk, the follow-up quality and the side-effects. RESULTS: During four months, 189 (14.3%) of 1317 medical patients have received a prophylaxis with LMWH; sixty one per cent of them were older than 70 years. Thrombotic risk as measured with Thilly's score was low in 50 patients (26%), moderate in 81 patients (43%), high in 58 patients (31%). Platelet count follow-up was optimal in 88 patients (47%). A decrease of platelet count over 30%, reaching less than 100 G/l, was recorded in 4 patients. A venous thrombosis was diagnosed clinically in one patient. Two patients had an overt severe bleeding. A serious hidden bleeding was suspected in 11 patients. CONCLUSION: LMWH were frequently prescribed for the prophylaxis of venous thromboembolic disease in medical patients. Most of these patients were over 70 years of age. Platelet count follow-up was in accordance with guidelines in less than half of the patients. Bleedings were not rare on this treatment. The present work suggests that a precise thrombotic risk assessment is needed before the onset of this therapy and that the association with aspirin should be careful. PMID- 11039084 TI - [An unrecognized neonatal emergency: extensive hematoma of the scalp. 5 cases]. AB - BACKGROUND: Subgaleal hemorrhage results from a pericranial effusion of blood subsequent to neonatal trauma. This exceptional situation compared with other pericranial effusion conditions in the neonate may be life-threatening. CASE REPORTS: We report the obstetrical and neonatal data in 5 cases of subgaleal hemorrhage observed in our unit over an 8-year 8-month period. We detail one particularly demonstrative case which illustrates the potentially serious course of certain clinical presentations. DISCUSSION: Subgaleal hemorrhage is a clinical diagnosis. Signs of hemorrhagic shock are associated with hemostasis disorders in the more severe forms of the condition. The main risk factor is instrumental delivery with suction. Careful monitoring is required. PMID- 11039086 TI - [Elevated salt taste detection threshold in subjects with essential arterial hypertension]. PMID- 11039085 TI - [Septic shock caused by Pasteurella multocida in alcoholic patients. Probable contamination of leg ulcers by the saliva of the domestic cats]. AB - BACKGROUND: Pasteurella multocida septicemic septic shock without scratches in chronic alcohol abusers are rare and paucibacillar. Chronic legs ulcers could be predisposing factors. Three severe cases of such diseases with early multiple organ failure without endocarditis despite 3, 5 and 3 positive blood cultures respectively are reported. CASE REPORTS: In all three cases, the source of septicemia was extensive skin legs ulcers contaminated by domestic cat saliva, probably. The underlying diseases were alcoholism, without cirrhosis (2 cases) and with cirrhosis (1 case). Aggravating factors were present (chronic renal insufficiency, diabetes mellitus, non-steroidal anti-inflammatory agents) and they might have been responsible for pejorative outcome despite early appropriate empirical antibiotic regimens. DISCUSSION: Because P. multocida SSC could be fulminant in chronic alcohol abusers with or without cirrhosis, we propose for these patients reducing exposure to cats and hand-washing after exposure if wounds are present. PMID- 11039087 TI - [Streptococcus A meningitis revealed by purpura fulminans]. PMID- 11039088 TI - [Nodule of the tongue revealing sarcoidosis]. PMID- 11039089 TI - [Lemierre syndrome secondary to malignant cervical adenopathy]. PMID- 11039090 TI - [Corticosteroid therapy and chronic obstructive bronchopneumopathies: the ISOLDE study]. PMID- 11039091 TI - [Remicade (infliximab) in the treatment of Crohn disease]. PMID- 11039092 TI - [To avoid complications, treatment of meningitis should follow the guidelines and be reevaluated at 48 hours]. PMID- 11039093 TI - [Acquired angioneurotic edema: association with hydatidosis]. PMID- 11039094 TI - [Corticosteroid therapy in obstructive lung diseases. Too much in COPD, not enough in asthma...]. PMID- 11039095 TI - [Corticosteroid therapy of asthma]. AB - BRONCHIAL INFLAMMATION AND GLUCOCORTICOIDS: Bronchial inflammation plays an important role in asthma and contributes to bronchoconstriction, hypersecretion and bronchial hyperreactivity. Glucocorticoids are the gold standard treatment in asthma affecting most of the components involved in bronchial inflammation. Inhaled steroids are recommended early in most countries. MECHANISM OF ACTION: The molecular and cellular mechanisms of glucocorticoids action are still better understood. However, it remains difficult to evaluate individual sensitivity when initiating treatment. Glucocorticoids have an effect on all inflammatory cells and bronchial structure cells. They bind to cytoplasmic receptors and then the complex links to DNA, inducing or inhibiting gene transcription in the target cell. DIFFERENT GLUCOCORTICOID SENSITIVITY: Very few patients are totally insensitive to the effect of glucocorticoids and require specific explorations for an identification. Although individual variability in corticosensitivity is similar in asthmatic patients and in the general population, the underlying mechanisms remain to be fully elucidated. The difference observed between responders and non-responders is not clearly identified and their definitions must be fully adapted. PMID- 11039096 TI - [Corticosteroid therapy of non-asthmatic chronic obstructive bronchopneumopathies]. AB - BASIS OF TREATMENT: Chronic obstructive pulmonary disease (COPD) is a common condition. Medical, and particularly drug, therapy still provides insufficiently effective relief. Corticosteroid treatment relies on the effect of these drugs on the underlying inflammatory mechanisms. Their efficacy has been demonstrated in asthma which exhibits certain features common with COPD. INDICATIONS: Short-term corticosteroid regimens are generally well tolerated. Clinical data favor their use in certain cases of acute decompensation. Long-term systemic regimens are not warranted due to the risk of adverse effects and the difficulty in maintaining appropriate dosages. Inhaled corticosteroids are widely used although the efficacy remains controversial. IMPORTANT DRAWBACKS: Clear evidence of efficacy from large controlled trials is still lacking. The difficulty encountered in obtaining such evidence is an indication of the minimal impact of such treatment and raises the question of its clinical pertinence. Patients exhibiting features similar to those observed in asthma (atopy, eosinophilia, improvement with bronchodilatation, non-smokers...) should be able to benefit from corticosteroids. For others a therapeutic test would be advisable to identify responders who could benefit from a preventive effect on the progression of COPD or associated asthma. A test lasting a few weeks at sufficient dosage is needed for subjective and objective (respiratory function tests) assessment. This costly therapy would not be warranted in non-responders, particularly in light of the expected secondary effects. Current evidence does not point to corticosteroid therapy as the much needed fully effective treatment for COPD. PMID- 11039098 TI - 13th Congress of the European College of Neuropsychopharmacology. Munich, Germany, September 9-13, 2000. Abstracts. PMID- 11039097 TI - [Level 2 analgesics in pain management: the rheumatologist's viewpoint]. PMID- 11039099 TI - Experimental Biology 2000--Milestones and Goals. Society for Experimental Biology annual symposium and joint international meeting. Cambridge, United Kingdom. 30 July-3 August 2000. Abstracts. PMID- 11039100 TI - American and European Associations for Cancer Education joint meeting. November 2 5, 2000. Washington, DC, USA. Abstracts. PMID- 11039101 TI - 2000 International Meeting of the Institute of Human Virology: a symposium on HIV/AIDS and related topics. September 10-15, 2000. Baltimore, Maryland, USA. Abstracts. PMID- 11039102 TI - XIth International Vascular Biology Meeting. Geneva, Switzerland, September 5-9, 2000. Abstracts. PMID- 11039104 TI - Joint meeting of the Nutrition Society, the Association for the Study of Obesity, and the British Hyperlipidaemia Association. 5-6 October, 1999. London, United Kingdom. Abstracts. PMID- 11039103 TI - [49th Meeting of the Association of North-German Ophthalmologists. Hamburg, 8-9 July 2000. Abstracts]. PMID- 11039105 TI - [Regional Research Conference of Psychiatric Hospitals of Bavaria. 28-29 October 1999. Abstracts]. PMID- 11039106 TI - [5th Congress of the German Society of Gerontology and Geriatrics. Nurnberg, 18 20 September 2000. Abstracts]. PMID- 11039107 TI - [51st Annual meeting of the German Society of Neurosurgery. 13-16 September 2000, Lubeck. Abstracts]. PMID- 11039108 TI - Guy Gordon Studdy Dutton. 1923-1997. PMID- 11039109 TI - Iodine in carbohydrate chemistry. PMID- 11039110 TI - Structure of anomeric glycosyl radicals and their transformations under reductive conditions. PMID- 11039111 TI - Oligosaccharide-protein conjugates as vaccine candidates against bacteria. PMID- 11039112 TI - Molecular structure of the carbohydrate-protein linkage region fragments from connective-tissue proteoglycans. PMID- 11039113 TI - The conformation of C-glycosyl compounds. PMID- 11039114 TI - Preventing complications of electromyography. AB - Complications of standard clinical electromyography and nerve conduction studies are reviewed, with suggestions on avoiding them. PMID- 11039115 TI - F-waves and conduction velocities range. AB - A controversial aspect in F-wave studies is if these potentials are generated preferentially by large motoneuron or by motoneuron of all sizes. The purpose of this work is to compare the maximum and minimum conduction velocities of the fibers that generate the M-wave with the maximum and minimum conduction velocities of the F-waves elicited by ulnar nerve stimulation. There were no significant differences between maximum velocities. However, minimum F-wave velocity was significantly higher than minimum conduction velocity, suggesting that the F-waves registered were preferentially generated by the fastest conducting axons. PMID- 11039116 TI - Human motor unit activity during concentric and eccentric movements. AB - Single motor units (MUs) activity was investigated in human m. biceps brachii during movements against an elastic load. A total of sixty-five MUs were studied by means of subcutaneously placed fine-wire branched electrodes. Subjects were asked to perform active shortening and lengthening of the muscle with approximately constant velocities at two different speeds--slow and fast. Both recruitment (RT) and decruitment (DT) thresholds of MU were found to be lower in movement with higher velocity. The recruitment order of MUs was approximately one and the same during concentric movements with a different but constant velocity. The firing onset of MUs is organized so that the peak of the first twitch contraction occurs at approximately the same force level irrespective of how fast the movement is. In contrast, during the eccentric movements the peak of the last twitch contraction of MU occurs at different torque levels depending on the velocity. The decruitment of the MUs during eccentric movement was in a reverse order to their recruitment during concentric movements. Generally, at one and the same velocity the RT of a given MU was lower than DT. Nevertheless, the peaks of the first and the last twitch contractions during concentric and eccentric movements with one and the same velocity occurred at approximately one and the same torque level. PMID- 11039117 TI - Jitter analysis utilizing a high speed FM tape recorder. AB - Jitter analysis in single fiber EMG (SFEMG) is usually done on-line during recording. However, this technique frequently prolongs the study and makes re analysis impossible. We attempted to measure jitter with a high speed FM tape recorder and compare the results with the previously published values. SFEMG data, acquired with voluntary activation on extensor digitorum communis muscle of 25 healthy relatives of children with myasthenia gravis were retrospectively analyzed. Fiber density (FD) was estimated on-line. Five to 18 single fiber action potential (SFAP) pairs were studied in each subject. The wow of the tape recorder was 6 microseconds. Mean (SD) (upper 95th percentile) FD, individual jitter, highest jitter, mean jitter and interspike interval were 1.60 (0.18) (1.90), 25.30 (11.20) (57.00) microseconds, 31.24 (6.87) (47.00) microseconds, 25.08 (5.04) (43.00) microseconds, and 0.67 (0.11) (0.91) ms respectively. Mean jitter in the pooled SFAP pairs and mean MCD were found to be lower than the published values of the Ad Hoc Committee of the AAEM Special Interest Group on Single Fiber EMG. A high speed FM tape recorder can be reliably used for the off line analysis of jitter. PMID- 11039118 TI - Electromyographical study of the iliocostalis lumborum, longissimus thoracis and spinalis thoracis muscles in various positions and movements. AB - The iliocostalis lumborum, longissimus thoracis and spinalis thoracis muscles were studied electromyographically in six male individuals between 18 and 23 years old. They were connected to co-axial needle electrodes while in orthostatic, kneeling and sitting positions performing movements of flexing, extending and rotating the trunk. In the total flexing of the trunk the muscles did not present any action potential. The results showed intense potential for action while flexing the trunk 45 degrees, extending the trunk beginning at 45 degrees of flexing and in homolateral rotation for the muscles analyzed in the orthostatic position, emphasizing the iliocostalis lumborum muscle in the extension of the trunk which registered very strong action potentials in all individuals. There were similar results for movements of flexing and extending the trunk in the kneeling position, emphasizing the longissimus thoracis muscle in the movement of hyperextension. In the sitting position the more intense potentials were for the movements of extension, flexing with rotation and homolateral rotation of the trunk, emphasizing the longissimus thoracis muscle with strong potentials. PMID- 11039119 TI - Study of the explosive strength of the rectus femoris muscle using electromyography. AB - The rectus femoris muscle was electromyographically studied in eleven individuals of the masculine sex. The objective was to study the action potential and the explosive strength of this particular muscle in the movement utilized to kick the field soccer ball and in other exercises of the orthostatic position. The following movements were analyzed: flexibility of the hip with the knee bent, flexibility and extension of the knee (completely crouched), flexibility of the hip with the knee extended, movement of the kick without contact with the ball and movement of the kick in contact with the ball. The results were: the rectus femoris muscle showed much stronger action potential in flexing the hip with the knee flexed more than 90 degrees; with the knee extended 60 to 90 degrees; in the movement extending the knee from a complete crouch and at the moment of the kick in contact with the ball. The action potential was strong in the movements flexing the knee (crouch) and in the initial movement of the kick. PMID- 11039120 TI - Deficit in sensory motor processing in depression and Alzheimer's disease: a study with EMG and event related potentials. AB - Event related potentials have been examined in depression and Alzheimer disease like clinical utility. To evaluate the influence of visual and auditory stimuli on the P300 latency we studied 12 patients with major depression, 12 patients with Alzheimer disease and 12 normal subjects. The experimental tasks applied was, first a series of 300 auditory stimuli, 255 (85%), with tones of 1,000 Hz, and considered as the frequent stimulus, whereas 45 (15%) were tones of 2,000 Hz and referred as the rare stimulus. A second series of 300 visual stimuli, 255 (85%) that were black circles on a white background, and considered the frequent stimulus (9 cm diameter, 200 ms duration), whereas 45 (15%) were black squares on a white background and referred as the rare stimulus (9 cm diameter, 200 ms duration) in the centre of a computer screen. The results show an increase of P300 latency in depressive and Alzheimer patients during auditory and visual tasks. Differences were found in reaction time to visual or auditory stimuli in Alzheimer disease. These results are consistent with an impairment in brain function in depressive patients that is associated with cortical hypoactivity and deficits in perceptive, auditory or visual, functions, whereas deterioration in Alzheimer's disease is sensorymotor, according to the slowness latency in the reaction time. PMID- 11039121 TI - Effect of ankle position fixation on peak torque and electromyographic activity of the knee flexors and extensors. AB - The purpose of this study was to examine the effect of ankle position fixation on peak torque (PT) and electromyographic (EMG) activity of knee-joint muscles during isokinetic testing. Twelve female athletes performed isokinetic knee flexion and extension at 60 degrees and 180 degrees/s under two conditions: with the ankle fixed in a position of plantarflexion and with the ankle fixed in a position of dorsiflexion. Bipolar surface electrodes were placed on the vastus lateralis, vastus medialis, biceps femoris, medial hamstrings, and the lateral head of the gastrocnemius for determination of the root mean square of the EMG (rmsEMG) and the median frequency of the EMG (mfEMG). No significant differences in knee extensor PT were noted in either ankle position for each velocity tested. Significant differences were noted, however, in knee flexor PT (p < 0.05) at both 60 degrees and 180 degrees/s, with the greatest PT observed with the ankle fixed in dorsiflexion. Neither quadriceps, hamstrings, nor gastrocnemius rmsEMG activity was affected by ankle position; however, there was a significant difference in mfEMG for the gastrocnemius, with higher frequencies observed with the ankle fixed in plantarflexion (p < 0.01). These results suggest that ankle position effects knee flexor PT during open chain isokinetic movements. The reason for decreased knee flexor PT with the ankle fixed in plantarflexion is probably due to the gastrocnemius muscle being in a too shortened position, thereby preventing it from effectively producing force at the knee joint. PMID- 11039122 TI - Normal values of F wave in lower extremities of 73 healthy individuals in Iran. AB - F wave latency has been shown to be a simple and valuable method in evaluation of proximal part of peripheral nerves. According to our previous study of F wave of upper extremity nerves (1), maximum normal F wave latency for the median nerve was 28 ms with stimulation at wrist and 25 ms with stimulation at elbow. These values for the ulnar nerve were 29 ms and 25 ms respectively. Maximum normal difference between right and left F wave latency with wrist stimulation was 2 ms for median nerve and 2.5 ms for ulnar nerve. Maximum normal difference between median and ulnar nerve F latency was 3.5 ms with stimulation at wrist. In this study we measured F wave of lower extremity nerves in 73 healthy individuals in Shiraz. Maximum normal F wave latency for tibial nerve was 55 ms with stimulation at ankle and 46 ms with stimulation at popliteal area. Maximum normal F wave latency for the peroneal nerve was 54 ms with stimulation at ankle and 47 ms with stimulation at fibular head. Mean F ratio for both nerves was 1.29 with stimulation at knee. Maximum normal difference in F wave latency between right and left lower extremities was 3.5 ms with stimulation at ankle and 3 ms with stimulation at knee for the peroneal nerve. These values were 3 ms and 2.5 ms for the tibial nerve respectively. Maximum normal difference in F wave latency between tibial and peroneal nerve was 4 ms with stimulation at ankle and 3 ms with stimulation at knee. PMID- 11039123 TI - Rise time or rise rate: which should be a criterion for analysing motor unit action potentials? AB - We measured the rise time (RT) and rise rate (RR) of motor unit action potentials (MUAPs) when sharp sounds are heard in concentric EMG in the first dorsal interosseus muscle (FDI) and biceps brachii muscle (BIC) of normal subjects. MUAPs from FDI muscle with an RT of less than 500 microseconds were 85%, and those of more than 500 microseconds were 15%. In contrast, MUAPs from BIC muscle with an RT of less than 500 microseconds were 65%, and those of more than 500 microseconds were 35%. Distributions of the RR for FDI and BIC were also determined. MUAPs from FDI muscle with an RR more than 0.3 mV/ms were 98.3%, and those of less than 0.3 mV/ms were 1.7%. In contrast, MUAPs from BIC muscle with an RR of more than 0.3 mV/ms were 93%, and those of less than 0.3 mV/ms were 7%. We conclude it is better to use RR than RT when accepting MUAPs in clinical EMG, because even when sharp sounds are heard, MUAPs do not always have an RT of less than 500 microseconds. The use of RT and the sharpness of MUAPS therefore need to be reconsidered, or RR should be used in clinical EMG by automatic program. PMID- 11039124 TI - New insights into the pathogenesis of erythropoietic protoporphyria and their impact on patient care. AB - Erythropoietic protoporphyria (EPP, MIM 177000) is an inherited disorder caused by a partial deficiency of ferrochelatase (FECH) which catalyses the chelation of iron into protoporphyrin to form haem. The majority of EPP patients experience solely a painful photosensitivity whereas a small number of them develop liver complications due to the accumulation of excessive amount of protoporphyrin in the liver. EPP is considered to be an autosomal dominant disorder, however, with a low clinical penetrance. To date, a total of 65 different mutations have been identified in the FECH gene of EPP patients. Among the 89 EPP patients who carry a "null allele" mutation which results in the formation of a truncated protein, 18 of them developed EPP-related liver complications. None of the 16 missense mutations identified among 19 patients on the other hand, have been associated with liver disease (P = 0.038). The allelic constellation of an overt patient consists of a mutated FECH allele and a "low expressed" normal allele and that of an asymptomatic carrier, a combination of a mutated and a normally expressed FECH allele. The identification of the "low expressed" allele is facilitated by haplotype analysis using two single nucleotide polymorphisms, -251 A/G in the promoter region and IVS1-23C/T. At the current time when only partially effective therapies are available, the disclosures of both "null allele" and the "low expression" mechanisms will improve patient management. CONCLUSION: While covering the important clinical aspect of erythropoietic protoporphyria, this article emphasises the latest achievements in the molecular genetics of the disorder. PMID- 11039125 TI - Gastro-oesophageal reflux, sleep pattern, apparent life threatening event and sudden infant death. The point of view of a gastro-enterologist. AB - Sudden infant death, apparent life threatening events and regurgitation occur mostly during the first 6 months of life, making it very tempting to suspect a relation between these phenomena. According to epidemiological data, the role of gastro-oesophageal reflux in the aetiology of apparent life threatening events is unclear. It has been demonstrated that sudden infant death is decreased in the supine versus the prone sleeping position. On the contrary, gastro-oesophageal reflux is more pronounced in the supine than in the prone position, both in infants and in older children, both in infants with physiological and with pathological reflux. In the supine position, infants do sleep shorter, have more rapid-eye-movement sleep and have more arousals than in the prone position. CONCLUSION: It is thought that in the majority of infants, gastro-oesophageal reflux stimulates arousals and thus may well be considered as "protective" for rather than "provoking" sudden infant death. However, this hypothesis needs to be validated. PMID- 11039126 TI - Serum osteocalcin has limited usefulness as a diagnostic marker for rickets. AB - Serum alkaline phosphatase (AP), the bone fraction of which is secreted by osteoblasts, is elevated in rickets. Both normal and elevated levels of serum osteocalcin (OC), a bone-specific marker secreted by osteoblasts, have been reported in rickets. Expression of the OC gene is enhanced by 1,25 dihydroxyvitamin D (1,25(OH)2D) in experimental models. This study assessed serum OC levels in 14 controls and 41 patients with active rickets divided into a phosphopenic (n = 20) and a calciopenic (n = 21) group. Phosphopenic subjects were older (9.5 versus 5.7 years, P = 0.03) with higher median serum calcium level (2.35 versus 2.16 mmol/l, P = 0.0002) and serum 25-hydroxyvitamin D level (15.4 versus 10.4 ng/l, P = 0.003); and lower serum phosphate (0.80 versus 1.51 mmol/l, P = 0.0001), serum 1,25(OH)2D (43.0 versus 95.6 pg/ml, P = 0.0001) and intact serum parathyroid hormone level (45.0 versus 141.5 ng/l, P = 0.01) than calciopenic subjects. There were no differences in median serum AP (774 versus 1430 IU/l, P = 0.17) and OC (14.5 versus 13.4 ng/ml, P = 0.6) between the two groups. The mean OC value for the 41 rickets subjects was 15.1 +/- 6.2 ng/ml and 17.4 +/- 7.8 ng/ml for the 14 control subjects. In the face of markedly elevated serum AP levels in the rickets subjects, all of the serum OC values in the study fell within two standard deviations of the mean for normals. There was no association between serum OC and 1,25-(OH)2D in either the phosphopenic or the calciopenic group. CONCLUSION: These results show that serum osteocalcin levels are not elevated in all forms of active rickets and that, unlike serum alkaline phosphatase, serum osteocalcin cannot be used in the diagnosis of rickets. PMID- 11039127 TI - Snoring, noisy breathing in sleep and daytime behaviour in 2-4-month-old infants. AB - The study aimed at evaluating possible associations between snoring and/or noisy breathing in sleep and daytime behaviour in 2-4-month-old infants using the Early Infancy Temperament Questionnaire (EITQ) as a tool. It covered the period from 1997 to 1998 and comprised 200 randomly selected clinically healthy infants aged 2-4 months from the community who were singletons and born in St. Petersburg within the chosen period. The mothers were asked to complete the questionnaires addressing infant, maternal, and major demographic characteristics, some infant care practices as well as the infant's habitual breathing symptoms during sleep. As a part of interview, the mothers answered the EITQ consisting of 76 items which describe different aspects of infant behaviour. Groups of questions were added according to a scoring sheet to produce total scores describing nine different aspects of infant temperament: activity, rhythm, approach, adaptability, intensity, mood, persistence, distractibility and threshold. In 129 cases (64.5%), mothers reported no respiratory disturbances during sleep in their infants. Mothers of ten infants (5.0%) described their babies as habitual snorers; 48 babies (24.0%) were characterised as having other than snoring noisy breathing during sleep, and 13 (6.5%) habitually had both snoring and noisy breathing. Symptomatic infants were rated as being moodier when awake compared with asymptomatic ones and most moodiness was the feature of those infants who had both snoring and noisy breathing during sleep. These associations remained after adjustment had been made for major potential confounders. CONCLUSION: Snoring and noisy breathing during sleep, rather common symptoms in young infants, may be associated with specific behavioural disturbances, and moody infants should be investigated carefully for possible obscure respiratory problems. PMID- 11039128 TI - High frequency hearing loss in Ullrich-Turner syndrome. AB - A total of 38 patients with Ullrich-Turner syndrome underwent standard otological and audiometric evaluation as well as high frequency audiological tests. Some 26 (68.4%) patients had a history of middle ear infections, and ten (26.3%) had required otolaryngological surgery. Conventional audiometry (125-8000 Hz) demonstrated normal hearing in only 25 of the ears (33%); between 500-4000 Hz, 16 ears (21.0%) had a mixed type and eight ears (10.5%) had conductive hearing loss. High frequency audiometry (8-18 kHz) revealed sensorineural hearing loss in 98.7% of the ears. Our results for conventional audiometry are in accordance with the literature. CONCLUSION: The detection of a high prevalence of hearing loss in the high frequency range brings a significant new perspective to the pursuit of the aetiology of ear and hearing problems in Ullrich-Turner syndrome. This pathology seems to be a premature variant of presbycusis and it may underlie future hearing impairment which will come to clinical attention only after it progresses to conventional testing frequencies. While further studies are underway to evaluate this aspect, routine otological and audiological follow-up of patients with Ullrich-Turner syndrome is warranted from the time of diagnosis. PMID- 11039129 TI - Cutis tricolor: congenital hyper- and hypopigmented lesions in a background of normal skin with and without associated systemic features: further expansion of the phenotype. AB - The term cutis tricolor describes the uncommon co-existence of congenital hyper- and hypopigmented macules, in close proximity to each other, in a background of normal skin so far seen in a 17-year-old patient with various other congenital defects. The suggested explanation for this phenomenon is allelic twin spotting. We report on two boys, aged 6 and 11 years, with an unusual combination of three different degrees of pigmentation, one of whom had in addition, psychomotor delay, dysmorphic features, musculoskeletal abnormalities and subcortical and periventricular white matter high signal lesions on brain neuroimaging. In both cases a search for mosaicism in peripheral blood lymphocytes and cultured fibroblasts was negative. In contrast to the previously reported case, the two children had large streaks or patches of hyper- and hypopigmented skin lesions, in close proximity to each other, involving large areas of the body. The rest of the skin had a normal intermediate pigmentation. CONCLUSION: This combination of three degrees of pigmentation in association with systemic defects in one child and the lack of such association in the other confirms and further expands the clinical phenotype of cutis tricolor. PMID- 11039130 TI - Prospective evaluation of late effects after childhood cancer therapy with a follow-up over 9 years. AB - Intensive multimodality treatment has led to a remarkable improvement of prognosis in paediatric cancer patients, however, a great number of long-term survivors suffer from considerable tumour- or treatment-related late effects. Between January 1990 and December 1998, 223 consecutive survivors of childhood malignancies entered a prospective follow-up study designed to evaluate the frequency and severity of tumour- and/or therapy-related long-term sequelae. After cessation of therapy and subsequently once a year, all patients underwent a detailed examination programme including physical examination, laboratory tests, abdominal sonography, echocardiography, electrocardiography, electroencephalography, spirometry, audiometry, ophthalmological examination and endocrine stimulation tests. Median follow-up was 5 years (range 0.4 to 9.6 years). A total of 167 patients (75%) had at least one chronic medical problem of whom 80 needed permanent medical support. The organ systems most frequently affected were the nervous system in 39%, the endocrine system in 32%, the ears/eyes in 22%, the kidneys in 17%, and the liver in 12% of the patients. Some late effects (endocrine deficits, hearing loss, tubulopathy) were primarily diagnosed only several years after the end of oncological therapy. CONCLUSION: The results of this study indicate that a considerable number of former paediatric cancer patients suffer from remarkable long-term side-effects. Since life quality is an important parameter of cancer survival, careful follow-up of long-term survivors is mandatory with the aim to reduce or even abrogate possible side-effects at the earliest time. PMID- 11039131 TI - Preventive effect of 2 and 10 mg of sodium cromoglycate on exercise-induced bronchoconstriction. AB - This double-blind, randomised and cross-over study was designed to compare the preventive effect against exercise-induced bronchoconstriction (EIB), defined as the percentage decrease in FEV1 > or = 15% after 6 min of exercise, of 2 mg and 10 mg of sodium cromoglycate (SCG), administered through a metered dose inhaler via spacer, in asthmatic children. Each of the 30 subject (age 11.6 +/- 3.2 years) was tested on five occasions. For inclusion, EIB in test1 was required. In tests 2 to 5, all subjects inhaled 2 mg or 10 mg of SCG 20 min and 120 min before exercise in a randomised order. In order to assess excretion of eosinophil protein X (EPX) accompanying EIB, urine samples were collected before and after exercise. The mean percentage fall in FEV1 (+/- SD) in test 1 was 26.8 +/- 9.8%. Inhalation of 2 mg and 10 mg of SCG 20 min before exercise provided a significant preventive effect in 83% and 77% and inhalation 120 min before exercise provided a preventive effect in 63% and 70%, respectively (n = 30). Variance analysis did not reveal a statistically different absolute fall in FEV1 after exercise when both doses (120 min before exercise) were compared (P = 0.356). In an unselected subgroup of 12 children, urinary EPX increased after the challenge without SCG premedication (test 1) (mean change: +48.7 micrograms/mmol creatinine, P = 0.034), whereas no significant increase was found in case of SCG premedication (mean change in microgram/mmol creatinine): 2 mg/20 min: +12.1; 2 mg/120 min: +8.5; 10 mg/20 min: -10.4 and 10 mg/120 min: -23.5; P > 0.1). CONCLUSION: Administration of 10 mg of sodium cromoglycate is no more effective in preventing exercise-induced bronchoconstriction than 2 mg regardless of whether the medication is given 20 or 120 min before exercise. The preventive effect of sodium cromoglycate on exercise-induced bronchoconstriction in asthmatic children is associated with the inhibition of urinary eosinophil protein X excretion. PMID- 11039132 TI - Improved dysgammaglobulinaemia in congenital rubella syndrome after immunoglobulin therapy: correlation with CD154 expression. AB - A boy with congenital rubella syndrome developed dysgammaglobulinaemia with elevated serum levels of IgM. CD154 was not induced on his peripheral blood mononuclear cells when rubella virus RNA was detected in his throat swabs and peripheral blood by reverse transcriptase polymerase chain reaction. Following intravenous immunoglobulin therapy, improvement of immunoglobulin abnormalities, disappearance of rubella virus and normalisation of CD154 expression were demonstrated. CONCLUSION: These findings implicate the efficacy of intravenous immunoglobulin therapy for dysgammaglobulinaemia in congenital rubella syndrome and a role of CD154 for a prolonged virus infection. PMID- 11039133 TI - The effects of synthetic and natural surfactant on fluid balance in acute respiratory distress syndrome. AB - To evaluate the effect of different surfactants on fluid balance in respiratory distress syndrome, we studied 24 premature infants who were randomised to receive either natural or synthetic surfactant. Data were collected on ventilatory parameters, daily urine output, daily weight, fluid intake and serum electrolytes. Ventilatory requirements decreased more rapidly in babies receiving natural surfactant, with significantly greater reductions in mean airway pressure from 1 to 48 h and oxygenation index from 1-18 h (P < 0.05). There were no differences in fluid intake and serum electrolytes. Mean daily urine output was higher in the group receiving natural surfactant (87 ml versus 61 ml, P < 0.05). This group also had a greater weight loss from birth weight (-146 g versus -65 g, P < 0.05). CONCLUSION: Natural surfactant produces an earlier reduction in ventilatory requirements with an earlier diuresis. This should influence fluid management in respiratory distress syndrome. PMID- 11039134 TI - Inhibitory effect of meconium on pulmonary surfactant function tested in vitro using the stable microbubble test. AB - Meconium aspiration syndrome is related to mechanical obstruction of the airways and subsequent chemical pneumonitis. It has also been suggested that meconium causes inhibition of surfactant function. To assess its inhibitory effect on surfactant function in vitro, we used a stable microbubble (SM) test that was thought to reflect the adequacy of pulmonary surfactant. The mixtures were prepared by adding serial dilutions of human meconium to various concentrations of Surfactant-TA (Surfacten). The SM count at each concentration of surfactant significantly increased with the increasing concentration of surfactant. This shows that the SM test closely reflects the quantified function of surfactant. When various concentrations of meconium were added to the surfactant concentration of 0.05 mg/ml and 0.25 mg/ml, the SM test results were decreased even at low concentrations of meconium. Also the increase in the meconium concentration caused a decrease in the SM test result, which was dependent on the surfactant and the meconium concentration, accordingly. These results suggest that meconium inhibits surfactant function. CONCLUSION: The stable microbubble test is an effective indirect method that tests the changes in surfactant quantity. In the in vitro experiment, we observed an inhibitory effect of meconium on the surfactant activity using the stable microbubble test. PMID- 11039135 TI - A randomised controlled trial of delayed cord clamping in very low birth weight preterm infants. AB - This study was carried out to assess the feasibility of late cord clamping of 45 s in preterm infants delivered mainly by caesarean section and the effects on postpartal adaptation and anaemia of prematurity. Prior to delivery, 40 infants of < 33 gestational weeks were randomised to either 20 s or 45 s of late cord clamping. After the first shoulder was delivered, oxytocin was given intravenously to the mother in order to enhance placento-fetal transfusion while the infant was held below the level of the placenta. The 20 infants in group 1 (20 s) had a mean birth weight of 1070 g and a mean gestational age of 29 + 4/7 weeks versus 1190 g and 30 weeks in group 2 (45 s). On day 42 of life there were ten infants without transfusions in group 2 versus three in group 1 (P < 0.05). Out of the 20 infants in group 1, 19 and 15/19 in group 2 were delivered by caesarean section. There were no significant differences in Apgar scores, temperature on admission, heart rate, blood pressure and requirements for artificial ventilation. CONCLUSION: Delayed cord clamping of 45 s is feasible and safe in preterm infants below 33 weeks of gestation. It is possible to perform the procedure at caesarean section deliveries and it should be performed whenever possible. It reduces the need for packed red cell transfusions during the first 6 weeks of life. PMID- 11039136 TI - Effectiveness of antiretroviral therapy initiated before the age of 2 months in infants vertically infected with human immunodeficiency virus type 1. AB - The effectiveness and tolerance of antiretroviral therapy with a combination of three reverse transcriptase inhibitors starting at the time of diagnosis (before 2 months of age) was evaluated in four infants with vertically acquired HIV-1 infection. Plasma HIV-1 RNA levels ranged from 230,000 to 1,000,000 copies/ml before onset of triple therapy and fell below 50 copies/ml at 12 to 33 weeks of life in three of the infants. These three children, currently aged 158, 105 and 72 weeks, are asymptomatic, have normal lymphocyte subsets and no hypergammaglobulinaemia. Two children experienced a profound reduction in the amount of proviral DNA detected in blood and have become HIV-1 seronegative, although one of them has had HIV-1 RNA detectable on a single occasion at 114 weeks of life (303 copies/ml). Transient interruption of therapy resulted in a rapid but reversible increase in HIV-1 RNA levels in the third child and was associated with the production of HIV-specific antibodies. The fourth child whose parents were not compliant to treatment and follow-up had a poor virological response. CONCLUSION: Early treatment of vertically acquired human immunodeficiency virus type 1 infection with three reverse transcriptase inhibitors is well tolerated and can result in such suppression of viral replication that specific antibodies are not produced, that proviral DNA falls to the lower limit of quantitation in blood and that all clinical and immunological manifestations of infection are avoided. Parental adhesion is crucial to the effectiveness of therapy. PMID- 11039137 TI - A child with dehydration and severe hypernatraemia. PMID- 11039138 TI - Varicella zoster virus induced haemolytic crisis in a child with congenital spherocytosis. PMID- 11039139 TI - Oral mucosal immunocompetence in preterm infants in the first 9 months of life. PMID- 11039140 TI - Can cord blood be used for autologous transfusion in preterm neonates? PMID- 11039141 TI - Appreciation expressed for article on cultural diversity. PMID- 11039142 TI - What does a patient's outfit weight? PMID- 11039143 TI - Can we make it better? PMID- 11039144 TI - Knowing what a human life really is: doctors and priests. PMID- 11039145 TI - Mini-ethnography: meaningful exploration made easy. PMID- 11039146 TI - Symptoms as a source of medical knowledge: understanding medically unexplained disorders in women. AB - In cases where a traditional medical frame of reference does not fully offer an adequate tool for understanding, the resident may find it perplexing to apply knowledge from medical school, where disease implies symptoms, findings, diagnosis, and cure. Medically unexplained disorders, mostly occurring in women, are chronic and disabling conditions, presenting with extensive subjective symptoms, although objective findings or causal explanations are lacking. Acquiring the knowledge and skills needed for adequate care of patients with chronic fatigue or pain syndromes is not an easy task. This paper presents a strategy for teaching, intended to facilitate understanding through appreciation of symptoms as a source of knowledge. The clinical approach is based on empowering practices supported by theoretical perspectives about signs, narratives, knowing, and gender. Also examined is the impact of commonly occurring teaching traps related to gender, psychosocial labeling, universalistic understanding, omnipotence, and power. PMID- 11039147 TI - The comfort of family practice residents with health care of patients of the opposite gender. AB - BACKGROUND AND OBJECTIVES: Studies have shown that female and male residents see disproportionately few patients of the opposite gender. Such an imbalance may be detrimental to the quality of residency training. This study examined residents' levels of comfort in caring for patients of the same and opposite genders and their assessments of the adequacy of the number of same- and opposite-gender patients they saw during residency training. METHODS: Data were collected from four family practice residency programs in the Midwest. All current family practice residents at the four sites surveyed were given a questionnaire asking them to rate their perceptions of the gender distributions of their patient panels and comfort with several areas of health care, some gender specific. The questionnaire used a five-point Likert scale. RESULTS: Ninety-four surveys (83% response rate) were collected. Ninety-three percent of female residents reported seeing enough female patients, but only 54% reported seeing enough male patients. Seventy-six percent of male residents reported seeing enough male patients, but only 31% reported seeing enough female patients. More females than males (88% versus 48%) felt comfortable with women's health, but more males than females (70% versus 31%) felt comfortable with men's health. Male residents were significantly more comfortable than female residents in performing or managing prostate exams, testicular exams, and prostatitis. Female residents were significantly more comfortable with clinical breast exams and breast disease. CONCLUSIONS: Male and female residents perceive disproportionate experiences in their exposure to patients of the opposite gender. Significant differences between male and female residents were also apparent regarding their relative comfort in managing some gender-specific health care topics. These results have implications for residency education. PMID- 11039148 TI - Differences in ambulatory teaching and learning by gender match of preceptors and students. AB - BACKGROUND AND OBJECTIVES: Important differences exist in traditional medical education by gender of the teachers and learners. Much less is known about how gender influences educational experiences in community-based ambulatory settings. In this study, we explored how community-based teaching and learning varies by gender of the students and preceptors. METHODS: This prospective study used both paper- and computer-based documentation systems to collect information on student patient-preceptor encounters. A consecutive sample of third-year medical students contributed data on one full clinical day each week as they rotated through a required 8-week family medicine clerkship. The main measures of interest included patient age and gender, health care visit type (acute, acute exacerbation of chronic, chronic, and health maintenance), method of learning in history taking and physical examinations (observing preceptor, being observed by preceptor, performing unobserved, or working jointly with preceptor), content of physical examinations, amount of preceptor feedback, preceptor teaching content, and gender of the students and their preceptors. RESULTS: Ninety-three students contributed data on 5,017 patient encounters. The distribution of encounters by student-preceptor dyad included: 1,926 (38%) female students with male preceptors. 1,716 (34%) male students with male preceptors, 841 (17%) female students with female preceptors, and 534 (11%) male students with female preceptors. We found that female preceptors conduct more complete physical exams with students than do male preceptors (28% versus 23%). Female students with male preceptors devoted more encounters to observation only than any other dyad (20% versus 12%), and female preceptors are more likely than male preceptors to allow students to perform unobserved (70% versus 59%). Patient gender played little if any role in how students and their preceptors worked together. CONCLUSIONS: Differences of potential importance were found in teaching and learning by gender of the student-preceptor dyad. This factor can and should be considered when determining how students can best meet educational objectives in community-based ambulatory settings. PMID- 11039149 TI - Gender and power in family medicine education. PMID- 11039150 TI - Dialogues in the exam room: medical interviewing by resident family physicians. AB - BACKGROUND: Limited information exists about the application of the biopsychosocial model in medical practice. This study expanded our knowledge about the extent to which psychosocial content is included in medical interviews conducted by resident family physicians. METHODS: Interviews of 180 patients conducted by six second-year family practice residents were audiotaped and transcribed. Physician statements were analyzed and coded as social talk, physician-centered statements, patient-centered statements, and discussion of patient affect, family, health promotion, and patient education. RESULTS: The proportion of interviews in which specific physician interactions occurred were physician-centered statements: 100%, patient-centered statements: 66%, dealing with patient affect: 18%, information about family: 61%, initiation of health promotion: 33%, and initiation of patient education: 46%. Discussions of patient opinion/perception, patient affect, family information, and health promotion occurred most commonly during well-care visits and with female patients. CONCLUSIONS: In this sample of residents, providers extended the interview beyond a purely biomedical focus. However, the psychosocial focus often was brief and applied inconsistently across patients. PMID- 11039151 TI - Alcohol-free instant hand sanitizer reduces elementary school illness absenteeism. AB - BACKGROUND AND HYPOTHESES: A substantial percentage of school absenteeism among children is related to transmissible infection. Rates of transmission can be reduced by hand washing with soap and water, but such washing occurs infrequently. This study tested whether an alcohol-free instant hand sanitizer (CleanHands) could reduce illness absenteeism in school-age children. METHODS: A 10-week, open-label, crossover study was performed on 420 elementary school-age children (ages 5-12). Students were given a brief orientation immediately prior to the start of the study on the relationship of germs, illness, and hand washing. Each student in the treatment group then received the test product in individual bottles, with instructions to apply one to two sprays to the hands after coming into the classroom, before eating, and after using the restroom, in addition to their normal hand washing with soap and water. The control group was instructed to continue hand washing as normal with non-medicated soap. After 4 weeks of treatment and a 2-week wash-out period, the control and experimental groups were reversed. Data gathered on absenteeism were classified as gastrointestinal or respiratory related and normalized for nonillness-related absenteeism and school holidays. RESULTS: Compared to the hand washing-only control group, students using CleanHands were found to have 41.9% fewer illness related absence days, representing a 28.9% and a 49.7% drop in gastrointestinal- and respiratory-related illnesses, respectively. Likewise, absence incidence decreased by 31.7%, consisting of a 44.2% and 50.2% decrease in incidence of gastrointestinal- and respiratory-related illnesses, respectively. No adverse events were reported during the study. CONCLUSIONS: Daily use of the instant hand sanitizer was associated with significantly lower rates of illness-related absenteeism. PMID- 11039152 TI - Association of attending physician specialty with the cesarean delivery rate in the same patient population. AB - BACKGROUND AND OBJECTIVES: In the context of a dramatic increase in US cesarean delivery rates over the past 30 years and explicit national goals to decrease the cesarean rate, previous retrospective studies have shown that pregnant women cared for by family physicians may be less likely to undergo cesarean delivery, compared with patients cared for by obstetricians. METHODS: We conducted a retrospective chart review of 3,560 deliveries from the family practice service of a community-based family practice residency from 1986-1995, focusing primarily on cesarean delivery rates during two periods of time. During period 1 (n = 1,063), all attending were private practice obstetricians. After a transition period, all births were attended by family medicine faculty (period 2, n = 1,346). RESULTS: The total cesarean delivery rate declined from 16.7% in period 1 to 11.1% in period 2. Repeat cesareans declined from 8.5% to 2.9%. CONCLUSIONS: In this community-based residency, a change in the specialty of the attending physician was associated with a 34% decline in the cesarean delivery rate. The observed decline in the cesarean rate could not be accounted for by any change in patient demographics or secular trends in cesarean delivery rates. PMID- 11039153 TI - Responding to a natural disaster with service learning. AB - This article describes a service learning experience at the University of North Carolina School of Medicine. Responding to a nearby natural disaster, the curriculum was modified to allow 71 second-year medical students to provide needed aid to affected communities and also fulfill learning objectives important to their training. The description and the lessons learned provide an example of how service learning experiences can be implemented to enrich a curriculum. PMID- 11039154 TI - Socioeconomic differences in self-assessed health in a chronically ill population: the role of different health aspects. AB - We investigated the role that different health aspects play in the explanation of socioeconomic differences in self-assessed health. Socioeconomic differences in self-assessed health were investigated in relation to chronic disease, functional limitations, psychosomatic symptoms, and perceived discomfort/distress. In multiple logistic regression analyses, for three cutoff points of self-assessed health, significant socioeconomic differences in self-assessed health could be observed after adjusting for age and gender. After separate adjustment for each of the four health aspects, the analyses showed that for a health assessment as less-than-good and less-than-fair, psychosomatic symptoms were the most powerful explanatory factor. Perceived discomfort/distress proved to be the most powerful factor for a poor health assessment. We found that socioeconomic differences in self-assessed health could, to a large extent (72-80%), be explained through socioeconomic differences in the prevalence of the four types of health problems included in the study. For all cutoff points, objective health aspects accounted for a relatively small part of the socioeconomic variability in self-assessed health. More subjective aspects of health accounted for more of the variability. PMID- 11039155 TI - Racial differences in adolescents' perceived vulnerability to disease and injury. AB - This study examined gender and racial differences in adolescents' risk perceptions of major diseases and motor vehicle injury and whether these perceptions agree with national mortality rates and parental health history. Adolescent (N = 135; 55% African-American) boys and girls reported on their chances compared to other adolescents of developing specific diseases or experiencing a motor vehicle injury and their knowledge of parental health history. Logistic regression models revealed that girls' risk perceptions were similar to boys' ratings even though females are at less risk than males per national figures. Caucasian adolescents inaccurately perceived that they were at significantly greater risk than African-American peers for motor vehicle injury, stroke, cancer, and heart attack. Adolescents' knowledge of a father's diabetes was predictive of greater perceived vulnerability to diabetes. PMID- 11039157 TI - Individual latent growth curves in the development of marijuana use from childhood to young adulthood. AB - The present study was designed to examine the relationship between unconventionality and marijuana use over time. The sample for this paper consisted of 532 male and female participants interviewed during early adolescence, late adolescence, their early twenties, and their late twenties. Latent growth modeling was used. The findings indicated that (1) the influence of initial unconventionality (T2) on initial marijuana use (T2) was stronger for males, (2) unconventionality at T2 was not significantly related to overall rate of growth in marijuana use, and (3) change in unconventionality was related to overall growth rate of marijuana use. The implications of the findings for prevention and treatment are discussed. PMID- 11039156 TI - Chest pain and the treatment of psychosocial/emotional distress in CAD patients. AB - Treatment of psychosocial/emotional distress as a strategy for diminishing chest pain in such patients remains entirely unutilized in standard care. Sixty-three patients with known or suspected CAD were entered in an aggressive lifestyle modification program. Patients completed the Symptom Checklist 90-Revised (SCL90R) at the diagnostic interview session, at 3 and at 12 months. Statistically significant drops were observed on multiple scales of the SCL90R at both 3 and 12 months. An item from the SCL90R was used as a proxy for angina. Multiple measures of emotional distress at baseline were found to correlate with chest pain at baseline, but not a number of traditional cardiovascular risk factors. The chest pain item displayed improvement at both 3 and 12 months. Improvement on all scales of the SCL90R correlated with improvement in chest pain. It may be possible to control chest pain in some CAD patients with psychosocial interventions. PMID- 11039158 TI - The effect of accuracy of perceptions of dietary-fat intake on perceived risk and intentions to change. AB - Consumption of excess fat increases risk for many health problems and diseases. In the present study, 188 undergraduate students were studied to understand self perceptions of dietary-fat intake and the impact of those perceptions. Findings indicated that the majority of participants had inaccurate perceptions about the amount of fat in their diets. Further, compared to people who overestimated dietary-fat intake, people who underestimated fat intake had lower perceived risk of cancer, had lower intentions to change, and demonstrated less knowledge about the dietary-fat content of many foods. Findings suggest that this unrealistic underestimation of fat intake is a cognitive barrier to dietary change and people who underestimate dietary fat intake may require more intensive intervention to change their diets. PMID- 11039160 TI - Comments on the "Ergonomics Program Standard" proposed by the Occupational Safety and Health Administration. PMID- 11039159 TI - Implicit cognition and HIV risk behavior. AB - Implicit cognition theory differs from most other approaches to health behavior in that it emphasizes neurobiologically plausible and experimentally documented memory association processes rather than rational decisions, considerations of pros and cons, or beliefs. The present study of adults from a community population investigated the predictive effects of implicit cognition, as well as behavioral and personality variables (sensation seeking, hostility, conscientiousness, and polydrug use), on risky sexual behaviors (lack of condom use, sex after drug use, and multiple sexual partners). In addition, this study simultaneously investigated the predictors in both a high-risk and a low-risk sample. Results showed that the implicit cognition indicator was a significant, independent predictor of lack of condom use in the high-risk sample. Polydrug use and sensation seeking also had important predictive effects. The results encourage more research on implicit cognition in health behavior and further document links among drug use, personality, and HIV risk behavior. PMID- 11039161 TI - Commentary on the scientific basis of the proposed Occupational Safety and Health Administration Ergonomics Program Standard. PMID- 11039162 TI - Occupational pesticide exposure and semen quality among Chinese workers. AB - This study investigated the association between occupational pesticide exposure and semen quality among Chinese workers. Male workers, 32 who were exposed to organophosphate pesticides and 43 who were not exposed were recruited from two nearby factories and interviewed. Following a work shift, semen and urine samples were collected for pesticide metabolite analysis. Semen samples were analyzed for sperm concentration, percentage of motility, and percentage of normal structure. Within the exposed group, the mean end-of-shift urinary p-nitrophenol levels were 0.22 and 0.15 mg/L for the high- and low-exposure subgroups, respectively. Linear regression analysis of individual semen parameters revealed a significant reduction of sperm concentration (35.9 x 10(6) vs 62.8 x 10(6), p < 0.01) and percentage of motility (47% vs 57%, p = 0.03) but not percentage of sperm with normal structure (57% vs 61%, p = 0.13). Multivariate modeling showed a significant overall shift in the mean semen parameter. Occupational exposure to ethylparathion and methamidophos seems to have a moderately adverse effect on semen quality. PMID- 11039164 TI - Disability management: corporate medical department management of employee health and productivity. AB - This study describes a proactive in-house program for managing short-term disability (STD) in the workforce of a very large banking system. The goals of this program were to (1) minimize the personal and economic impacts of STD by early intervention, (2) validate the extent and duration of STD, and (3) coordinate medical services and provide guidance to managers that would facilitate an early return to work. This program was made possible by the installation of a comprehensive database, called Occupational Medicine and Nursing Information System. This database mainly includes employees' claims for inpatient and outpatient health services, disability and workers' compensation benefits, wellness program participation, medical examinations and laboratory tests, use of prescription drugs, and results of Health Risk Appraisals. As a result of these efforts, STD event duration declined after this STD management program was implemented in locations heretofore outside the system, and by providing full pay for part-time work after STD, within the system as well. Of note, the average number of STD days per employee showed substantial variation by health plan, including the fact that it was higher (3.9 STD days/employee) for health maintenance organization participants than for indemnity plan members (2.7 STD days/employee). PMID- 11039163 TI - Semen quality and hormone levels among radiofrequency heater operators. AB - Approximately 9,000,000 US workers are occupationally exposed to radiofrequency (RF) radiation; over 250,000 operate RF dielectric heaters. Our purpose was to determine whether male RF heater operators experience increased adverse reproductive effects reflected in reduced semen quality or altered hormone levels. We measured incident RF heater radiation exposures and RF-induced foot currents at four companies. For 12 male heater operators and a comparison group of 34 RF-unexposed men, we measured 33 parameters of semen quality and four serum hormones. Despite wide variation in individual exposure levels, near field strengths and induced foot currents did not exceed current standard levels and guidelines. We observed minor semen quality and hormonal differences between the groups, including a slightly higher mean follicle-stimulating hormone level for exposed operators (7.6 vs 5.8 mIU/mL). Further occupational studies of RF-exposed men may be warranted. PMID- 11039165 TI - Quantitative analysis of cumulative trauma risk factors and risk factor interactions. AB - This study identifies the strongest occupational risk factors and risk factor interactions in the development of indicators associated with cumulative trauma disorders (CTD). Suggestions for minimizing the occurrence of indicators associated with onset of CTD are presented. Five Occupational risk factors and six risk factor interactions were evaluated to determine their contribution to accepted indicators of the onset of CTD, including exertion, discomfort, difficulty, and fatigue. Taguchi's level-analysis procedure was used to design a 16-trial hammering experiment and to analyze the results. From the hypothesized occupational risk factors, repetition was found to have the strongest effect on the values of the physiological and subjective indicators of the onset of CTD. PMID- 11039166 TI - A technique to re-assess epidemiologic evidence in light of the healthy worker effect: the case of firefighting and heart disease. AB - The healthy worker effect (HWE) is a bias that is believed to have strongly affected the validity of previous cohort mortality studies on the relationship between firefighting and heart disease. There is a strong healthy hired effect (a component of the HWE) among firefighters, owing particularly to the recruitment of nondiabetic candidates. This is shown in previous studies in which the reported standardized mortality ratios for diabetes are much less than unity, generally around 0.3 to 0.5. Because diabetes is known to increase the risk of heart disease, a deficit of diabetes among firefighters is expected to lead to a deficit of heart injury and disease. This would make the cohort mortality studies incapable of detecting any increase in risk of heart injury and disease among firefighters. There is also a strong healthy worker survivor effect (another component of the HWE) among firefighters. In addition, heart disease is a classic example of the HWE because heart disease is chronic and its risk factors can be identified in the selection process. It is believed that (1) a major problem of previous studies on firefighting and heart disease is their failure to recognize the importance of the HWE when interpreting their results, and (2) a technique to re-assess results in light of the HWE is urgently needed. This article addresses the generally accepted principles relating to the HWE, including its definition and sources, and proposes a technique for re-assessing the literature in light of the HWE. The technique was applied to carefully re-assess 23 studies that provided direct evidence for the relationship between firefighting and heart disease. Before the re-assessment, 7 of the 23 studies showed positive evidence and 16 showed no evidence. After the re-assessment, 11 studies showed positive evidence and 12 showed no evidence. Based on the results of the re-assessment of the 23 studies, we concluded that (1) there is strong evidence of an increased risk of death overall from heart disease among firefighters; (2) there is insufficient evidence, even after considering the HWE, that there is an increased risk of death from aortic aneurysm among firefighters; and (3) there is insufficient evidence, even after considering the HWE, for a relationship between firefighting and any heart disease subtype, such as acute myocardial infarction. PMID- 11039167 TI - Risk of injury in African American hospital workers. AB - Very few data exist that describe the risk of injury in African American health care workers, who are highly represented in health care occupations. The present study examined the risk for work-related injury in African American hospital workers. Hospital Occupational Health Service medical records and a hospital human resource database were used to compare risk of injury between African American and white workers after adjusting for gender, age, physical demand of the job, and total hours worked. Risk of work-related injury was 2.3 times higher in African Americans. This difference was not explained by the other independent variables. Differences in injury reporting, intra-job workload, psychosocial factors, and organizational factors are all potential explanations for racial disparity in occupational injury. More research is needed to clarify these findings. PMID- 11039168 TI - [Is the independence of the medical press threatened?]. PMID- 11039169 TI - [Interferon and Behcet's disease. Therapeutic exception or loss of chance?]. PMID- 11039170 TI - [Clinical and prognostic aspects of spontaneous fractures in long term care units: a thirty month prospective study. Eastern Gerontology Society]. AB - PURPOSE: Spontaneous fractures (stress and bone insufficiency fractures) are well described in young healthy patients; however, few studies were conducted in the elderly. METHODS: A 30-month prospective clinical and epidemiological survey including elderly patients from long-term nursing homes (LTNH) of the Societe de Gerontologie de l'Est (70 centers; 11,495 elderly patients in total) was conducted. RESULTS: Sixty-seven spontaneous fractures were encountered in 30 LTNH (3,052 elderly patients) (five stress fractures of the foot, 62 bone insufficiency fractures). The mean age of bedridden patients was 85 +/- 7 years. The prevalence of spontaneous fractures (calculated from the number of patients admitted consecutively in LTNHs) was 0.34% in the whole population (11,495 beds). When the calculation was based on LTNH reports of spontaneous fractures (3,052 elderly patients), the prevalence reached 1.3%. Fractures of long bones were common in elderly patients and included 15 fractures of the femoral neck, 14 fractures of either the tibia or fibula, 13 fractures of the femoral shaft, and 11 fractures of the humerus. Fractures of the femoral shaft were associated with the highest mortality: seven out of 13 patients died versus two out of 15 patients with regard to fractures of the femoral neck (P < 0.05). CONCLUSION: Bone insufficiency fractures have not the same course in young healthy patients as those in elderly nursing home patients: they more often concern long bones and their prognosis is worse. Means of prevention still have to be defined. PMID- 11039171 TI - [Cutaneous reactions or necrosis from interferon alpha: can interferon be reintroduced after healing? Six case reports]. AB - PURPOSE: Alpha, beta or gamma interferon (INF) are cytokines produced by cells in response to antigenic stimulation. They are used to treat various hepatic, hematological, oncological and neurological diseases. Cutaneous reactions (rash, alopecia, labial herpes, erythema, or induration at the site of injection, and more rarely cutaneous necrosis) represent 5 to 12% of side-effects observed in patients receiving INF. The authors report six cases of local cutaneous reactions to alpha INF, five of which corresponded to cutaneous necrosis. This makes them question the relevance of INF reintroduction. METHODS: The study included 5 male and 1 female patients (mean age: 59.1 years; range: 42 to 74 years old). Three patients had chronic hepatitis C, while three others presented a blood disease. RESULTS: Cutaneous necrosis occurred after 1 to 10 months of treatment. The mean time to healing was 16.2 weeks. Reintroduction of the drug including injection in other sites did not lead to recurrence of necrosis in five out of the six cases. CONCLUSION: INF-induced cutaneous necrosis does not depend on the type of INF, the site of injection, the dose and may occur 2 months to 9 years after treatment implementation. The exact mechanisms involved in cutaneous necrosis remain unknown. Morbidity is due to a very long time to healing (4 to 6 months). Futhermore, healing sometimes requires prior surgery. Physicians should be aware of the potential occurrence of erythema in patients treated by INF, as it is the first sign of necrosis. The site of injection should then be modified. In case of necrosis, risk factors for thrombophilia, factors reducing microcirculation (DHE, beta-blockers, cigarette smoking) should be investigated. INF injections should be cautiously reintroduced in other sites with the help of a nurse in case of self-injections prior to the occurrence of necrosis. Regarding self-injections patients' training should be emphasized. PMID- 11039172 TI - [Cancer and the elderly. Management. Decision aspects]. AB - INTRODUCTION: Age is the major risk factor for the majority of patients with cancer. More than 50% of cancers occurs after the age of 60. Cancer in the elderly is therefore a public health issue at stake. However, in daily clinical practice the elderly presenting cancer are not listened to with great interest and treatment is often not proper or suboptimal. CURRENT KNOWLEDGE AND KEY POINTS: Diagnosis in the elderly is established at a more advanced stage of cancer than in younger people; diagnostic workup is reduced and suboptimal treatments are implemented. Therefore, barriers exist that prevent the elderly from accessing the healthcare system as easily as their younger counterpart. Misconceptions about cancer also lead them to delay their first visit. As well, although treatment with curative intent and without major side-effect is feasible, physicians have misconceptions regarding therapeutic possibilities. Due to the heterogeneity of the so-called "ageing population", difficulties are related to patients' selection. FUTURE PROSPECTS AND PROJECTS: Decision in oncology for the elderly must walk a fine line in attempting to deliver the best treatment under the best conditions. Age per se must not be the only criterion for medical decision. Providing accurate information adapted to the elderly, with large circulation among healthcare professionals, should lead to the same quality of care as that in young people. Comprehensive multimodal geriatric assessments should help to further differentiate patients who may benefit from curative treatment from those for whom only palliative treatment is necessary. PMID- 11039173 TI - [Idiopathic thrombotic thrombocytopenic purpura or Moschowitz syndrome: current physiopathologic and therapeutic perspectives]. AB - INTRODUCTION: The objective of this work was to review current data about the physiopathology, clinical features, and treatment of thrombotic thrombocytopenic purpura (Moschowitz's syndrome). CURRENT KNOWLEDGE AND KEY POINTS: Thrombotic thrombocytopenic purpura is a rare disorder characterized by widespread thrombotic injuries of platelets in the microcirculation. Its physiopathology has been elucidated recently. Evidence of a deficiency of Von Willebrand's factor cleaving protease would be due to either IgG antibodies in the acute form of the disease or constitutional deficiency in the chronic form of the disease. FUTURE PROSPECTS AND PROJECTS: Plasma exchange is the current reference treatment. However, in the light of recent publications, either infusions of concentrates of purified enzyme or more intensive immunosuppressive therapy would be more specific. PMID- 11039174 TI - [Systemic lupus erythematosus and risk of hepatitis B vaccination: from level of evidence to prescription]. AB - INTRODUCTION: Case reports focusing on immunological diseases occurring subsequently to vaccination are often described in the literature. Reporting of such cases may influence physicians' perception of risks related to immunization, and thereby immunization practices. The decision to vaccinate a patient with an immunological disease should not rely on such case reports, but on the level of evidence of a causal relationship between vaccination and the occurrence of an adverse event. This article describes the search for available data supporting such causality before taking the decision to introduce vaccination against hepatitis B in a female patient with systemic lupus erythematosus (SLE). CURRENT KNOWLEDGE AND KEY POINTS: Data extracted from Medline and surveillance system showed that: 1) biologic plausibility of a relationship between the HBs antigen and SLE was unlikely; 2) case reports or case series were seldom and not convincing regarding potential causality; and 3) there were neither controlled observational studies nor controlled clinical trials. The only available clinical study was of poor quality and did not show any adverse event. The level of evidence of a causal relationship between vaccination against hepatitis B and the occurrence of an adverse event in patients with SLE was low, in-between levels 4 and 5 as defined by the Center for evidence-based medicine. The risk-benefit ratio may therefore rely on these results and guide the decision whether or not vaccination should be introduced. FUTURE PROSPECTS AND PROJECTS: The type of reasoning reported in this paper can be used for other vaccines or other immunological diseases, and have wider applicability in terms of therapeutic risk management when data and evaluation are lacking that could guide decisions. PMID- 11039175 TI - [Activated C protein resistance manifested by cutaneous necrosis after interferon alpha injection: case report]. AB - INTRODUCTION: Cutaneous necrosis occurring in the course of treatment by alpha interferon is an uncommon side-effect. Its physiopathologic mechanism remains obscure. A local thrombotic action of interferon has been suggested to explain its occurrence. EXEGESIS: A 64-year-old male patient with human immunodeficiency virus-related cutaneous Kaposi's sarcoma presented cutaneous necrosis after a 9 month treatment by interferon alpha, while his resistance to activated protein C had already been demonstrated. To our knowledge, this is the first case ever described regarding the association of interferon-induced cutaneous necrosis with activated protein C resistance. CONCLUSION: This suggests that in case of interferon treatment-induced cutaneous necrosis coagulation disorders should be investigated and questions the existence of a particular "pro-coagulant profile" facilitating this side effect. PMID- 11039176 TI - [Cavernous sinus lymphoma mimicking Tolosa-Hunt syndrome]. AB - INTRODUCTION: Tolosa-Hunt syndrome is characterized by painful ophthalmoplegia due to idiopathic granulomatous inflammation of the cavernous sinus. Steroid therapy dramatically reverses the symptoms and clinical signs. Because they also may respond to steroids, tumors such as lymphoma and meningioma and orbital tumors can make differential diagnosis difficult. EXEGESIS: We report the case of a 78-year-old male patient in whom systemic lymphoma associated with inflammation of the cavernous sinus was uncovered by painful, gradually progressing, ophthalmoplegia mimicking Tolosa-Hunt syndrome. CONCLUSION: When faced with a clinical picture suggestive of the existence of Tolosa-Hunt syndrome clinical workup is mandatory and should lead to diagnosis of exclusion. PMID- 11039177 TI - [IgD myeloma manifesting as acute renal insufficiency]. AB - INTRODUCTION: IgD myeloma is a rare disease, comprising only 1-2% of all cases of myeloma. EXEGESIS: A 71-year-old woman was admitted with acute renal failure, hypercalcemia and IgD lambda multiple myeloma. Dialysis was necessary. Six monthly cures of chemotherapy of induction according to the protocol VAD (vincristine, doxorubicin and dexamethasone) allowed to achieve moderate chronic renal failure (serum creatinine = 120 mumol/L). Sixteen months later, the patient developed an abdominal mass due to an IgD plasmocytoma in spite of treatment with interferon alpha and dexamethasone. Chemotherapy with melphalan and dexamethasone allowed to the disappearance of plasmocytoma and remission. The death occurred 36 months after the diagnosis. CONCLUSION: This observation allows to display the particularities of IgD myeloma: remarkable preponderance of lambda-type light chains, small or no visible monoclonal spike on serum electrophoresis, frequent extraosseous spread of tumor, renal failure and presence of osteolytic lesions. Over the last years, management and prediction of the survival time of IgD myeloma patients have improved. PMID- 11039178 TI - [An anterior thoracic painful cord]. PMID- 11039179 TI - [Too much skin, not enough blood!]. PMID- 11039180 TI - [Listeria monocytogenes: a rare cause of pleurisy]. PMID- 11039182 TI - Cumulative index volumes 34-36, 1996-1998 PMID- 11039181 TI - [A rare cause of spontaneous hemoperitoneum]. PMID- 11039183 TI - New tetrahydro-2H-1,3,5-thiadiazine-2-thione derivatives as potential antimicrobial agents. AB - The deacylated chloramphenicol amine D-(-)-threo-2-amino-1-(4-nitrophenyl)-1,3 diol (D-amine, 1a), and its enantiomer, the L-(+)-threo-form (L-amine, 1b), were introduced into a tetrahydro-2H-1,3,5-thiadiazine-2-thione (THTT) skeleton. They are incorporated in three ways (Chart 1, types I-III) at N3 (type I), N5 (type II) or both N3 and N5 (type III) of the THTT system. These selections were made in order to investigate the effect of combining the structural features of the THTT and the D-amine on the antimicrobial activity, if any. PMID- 11039184 TI - Azepino- and diazepinoindoles: synthesis and dopamine receptor binding profiles. AB - Starting from the readily available building blocks 7, 10, 11, and 15, the synthesis of the fused indoles 1, 2, 5, and 6, respectively, is reported. The syntheses involved Pictet-Spengler cyclizations, Michael addition reactions, lactamization, directed metallation, and reductive amination as the key reaction steps. Radioligand displacement studies comprising the dopamine receptor subtypes D1, D2long D2short, D3, and D4.4 were performed when the diazepinoindole 6 revealed D1 and D4 affinities (Ki = 0.11 microM and 1.7 microM, respectively) which are comparable to the partial D1 agonist SKF 38393 (3b). In contrast to the benzazepine 3b, the indole based test compounds turned out less selective over the D2 and D3 receptor subtype. PMID- 11039185 TI - Evaluation of methylthio-, methylsulfinyl-, and methylsulfonyl-analogs of alkanes and alkanoic acids as cardiac inotropic and antifungal agents. AB - A group of alkane and alkanoic acid compounds of general formula MeS(O)m(CH2)nR [m = 0-2; n = 1, 5, 13; R = Me, CO2H(Na)] were synthesized for evaluation as cardiac inotropic and antifungal agents. Inotropic activity was determined as the ability of the test compound to modulate in vitro guinea pig atrium contractility. The oxidation state of the S-atom was an important determinant of inotropic modulation since the thio (m = 0) analogs exhibited a positive inotropic effect. In contrast, the sulfinyl (m = 1) and sulfonyl (m = 2) analogs exhibited a negative inotropic effect. A pentyl spacer (n = 5) provided the largest positive or negative inotropic effect. The relative positive, and negative, inotropic potency orders with respect to the R-substituent were Me > or = CO2H, and CO2Na > or = Me, respectively. The most potent positive inotrope MeS(CH2)5Me (EC50 = 4.49 x 10(-6) M) could serve as a useful lead-compound for the design of a new class of positive inotropic agents. In a broad spectrum antifungal screen, the minimal inhibitory concentration (MIC) range for the five most active compounds was MeSO2(CH2)5Me (0.46-1.83 mM), MeS(CH2)13Me (0.31-1.23 mM), MeSO(CH2)13Me (< 0.009-1.87 mM), MeSO2(CH2)13Me (0.27-1.09 mM), and MeS(CH2)13CO2H (0.27-1.09 mM), relative to the reference drug Ampotericin B (< 0.0002-0.002 mM). The most active antifungal agent MeSO(CH2)13Me was selective against C. guillermondi, C. neoformans, S. cerevisiae, and A. fumigatus (strain TIMM 1776). PMID- 11039186 TI - A new class of antifungal agents. Synthesis and antimycotic activity of disubstituted N-azolylamines. AB - In this study we extended our exploration of the N-azolylamine moiety for its antifungal activity. We prepared a number of N-azolylamino derivatives. The synthetic sequence includes the preparation of aminoazole Schiff bases, and the reduction and the alkylation of the corresponding secondary amines. The title compounds were evaluated in vitro against several pathogenic fungi responsible for human disease. The most potent antimicrobial compound was the N-(biphenyl-4 yl)methyl-N-(2,4-dichlorophenyl)methyl-1H-imidazol-l-yl amine (21), which was found to be active against yeasts and dermatophytes; its potency and selectivity were comparable to those of miconazole. PMID- 11039187 TI - Antiestrogenic activities of 3,8-dihydroxy-6,11-dihydrobenzo[a]carbazoles with sulfur-containing side chains. AB - The objective of this study was to explore whether the conversion of the 2 phenylindole system into the tetracyclic benzo[a]carbazole changes the endocrine profile when the side chain structure was kept constant. Five different sulfur containing side chains were linked to the nitrogen of the tetracycle. The biological evaluation revealed that the character of the indole derivatives remained unchanged after the conversion to the respective benzocarbzoles but the potency decreased by one order of magnitude. In vitro, all derivatives acted as pure antiestrogens without any agonist activity. They strongly inhibited the growth of estrogen-sensitive MCF-7 breast cancer cells with IC50-values in the nanomolar range. In the mouse uterine weight test, the derivatives with an aliphatic side chain were devoid of estrogenic activity and antagonized the effect of estradiol. The presence of an aromatic ring in the side chain gave rise to significant agonist activity in vivo independently of the carrier structure. All data revealed the equivalence of both carrier structures in respect to the endocrine profile but showed a decrease in potency upon the conversion of the 2 phenylindole system into the benzocarbazole structure. PMID- 11039188 TI - Comparison of the inhibition of the cytosolic phospholipase A2-mediated arachidonic acid release by several indole-2-carboxylic acids and 3-(pyrrol-2 yl)propionic acids in bovine and in human platelets. AB - The inhibition of the cytosolic phospholipase A2 (cPLA2)-mediated arachidonic acid release by several indole-2-carboxylic acids and 3-(pyrrol-2-yl)propionic acids was measured in intact human platelets using calcium ionophore A23187 as stimulant. The comparison of the obtained data with the inhibition data evaluated with bovine platelets showed that analogous results were obtained with both cell types. PMID- 11039190 TI - A revival of Paul Dudley White an overview of present medical practice and society. P.D. White International Lecture PMID- 11039189 TI - Convenient procedures for synthesis of ciproxifan, a histamine H3-receptor antagonist. AB - Cyclopropyl 4-(3-(1H-imidazol-4-yl)propyloxy)phenyl methanone (ciproxifan) is a novel reference antagonist for the histamine H3 receptor. Despite the former Mitsunobu reaction the actual key reaction for preparation based on SNAr for acylated fluoroaromatics with an additional cyclization in a one-pot procedure needs no chromatographic purification steps and results in good yields. PMID- 11039191 TI - [The health care revolution in the USA]. AB - The changes in medical care in the USA have not been completed yet. The government which will be established after the following presidential elections will be compelled to deal with the organisation of medical care in order to make the financial participation of the federal government manageable. The American medical workers must unite in their procedures in order to bring the negotiations with the government institutions, insurers and health maintaining institutions to a successful end. The struggle for medical independence is of great importance as it enables to suppress stereotypes and obsolete traditions, and leads to innovative procedures which represent the basis for progress in medical science. (Fig. 10, Ref. 16.) PMID- 11039192 TI - The cosmos and CHAT, prompting blood pressure and heart rate monitoring for Derer's week. AB - BACKGROUND: The effects of natural environmental factors upon health, documented in Minnesota, support the proposition of Bratislava's champions of the cosmos and of the biological week, gauged via circaseptan rhythms by the late Ladislav Derer, whose "macro-rhythm" lasted "most frequently about 6 days". MAIN PURPOSE: To introduce 7-day monitoring of blood pressure and heart rate into routine practice. STARTING POINTS AND METHODS: Cosinor analysis on 7-day series determines (conventionally ignored) consistent blood pressure overswinging, i.e., circadian hyper-amplitude-tension (CHAT), a disease risk syndrome, whether it is associated with a normal average blood pressure or a high blood pressure. RESULTS: Summary of information understandable by the general population on the dynamics of blood pressure. CONCLUSIONS AND MEANING FOR PRACTICE AND THEORY: Space weather reports may prompt preventive measures. Caution dictates in any event monitoring blood pressure and heart rate for 7 days to attempt to prevent strokes, rather than to ignore the greatest yet detectable risk of catastrophic vascular disease, CHAT, a risk greater than old age or high blood pressure. (Tab. 1, Fig. 3, Ref. 31.) PMID- 11039193 TI - The remote past and near future of electrocardiology: view-point of a biomedical engineer. AB - The major steps of advancement in electrocardiology over a period exceeding one century of its existence are briefly summarized, and some considerations concerning the most promising trends of its current and future progress are presented. (Tab. 2, Fig. 2, Ref. 39.) PMID- 11039194 TI - [Detection of peptidergic and nitrergic structures in the spinal ganglia of rabbits]. AB - In this study we have demonstrated the presence of neuropeptide substance P and non-peptide neurotransmitter NO (nitric oxide) in the dorsal root ganglia of rabbit. NADPH-diaphorase histochemical staining was used for the detection of NO and immunohistochemical method for the detection of substance P.A particular number of dorsal root ganglion (DRG) cells were stained by SP and NADPH-d reaction. The presence of SP and NADPH-diaphorase positive cells varied depending upon spinal level of DRGs. Positively stained neurons were only small or medium sized. Cells of large diameter profiles showed no staining. Substance P immunoreactive cells were stained brown and dark brown, the intensity of NADPH-d staining varied from light to very dark blue. In some DRGs cells, there was a very significant neuronal co-localization of immunoreactivity for SP and reactivity for NADPH-d. In summary, DRG cells appear to express diaphorase and substance P activity, and some of them contain both neurotransmitters. Recent studies analysing the participation of NO in the regulation of SP release in the spinal cord suggest, that the DRGs neurons may display a close interaction between NO and SP. (Fig. 14, Ref. 39.) PMID- 11039195 TI - [Mineralized dental enamel matrix proteins]. AB - The organic matrix of the developing enamel is highly heterogeneous, comprising proteins derived from a number of different genes, including amelogenin, enamelin, ameloblastin, tuftelin, dentine sialophosphoprotein, enzymes and serum proteins. Each of these classes appears to undergo post-secretory sequential degradation which contributes towards further matrix heterogeneity. Possible functions of these proteins are discussed. Proteins of the organic matrix are presumed to play an important role in the modulation of mineral deposition and growth during tooth morphogenesis but the precise functions of individual matrix proteins remain unclear. (Tab. 1, Fig. 2, Ref. 47). PMID- 11039196 TI - [Laboratory diagnosis of toxoplasmosis]. AB - Toxoplasmosis is a world-wide spread parasitosis. The disease potentially highly affects two groups of patients: foetus and immunosuppressed patients. The determination of diagnosis and therapy on the basis of a single serum examination is very important; possible on the basis of a single serum sample. In most cases, it is possible to differentiate between recent and latent infections using a combination of suitable methods, which permit us to confirm particular antibody classes. In the presented paper the authors suggest diagnostic procedures for 4 groups of patients: pregnant, neonates with suspected congenital toxoplasmosis, immunodeficient patients and immunocompetent patients. The diagnostic methods consist of a combination of basic and supplemented diagnostic methods. Each patient's serum should be tested by basic tests which include the detection of total antibodies with CFT or IFT and specific classes of IgM and IgG antibodies by ELISA. The potential activity of toxoplasma infection can be determined by supplementary methods of e.g. IgG avidity antibodies, establishment of IgA antibodies, western blotting method and monitoring of antibodies production. For each situation the authors present interpretations of suspected cases including proposals for clinicians. These procedures are suggested for practical use in laboratories of various diagnostic levels in order to help to the diagnostic procedures in a particular situation as well as for clinical evaluation of established results. (Fig. 4, Ref. 65). PMID- 11039197 TI - [Diagnosis and treatment of functional disorders of the lower urinary tract in younger men]. AB - BACKGROUND: Functional disorders of the lower urinary tract can manifest by different symptoms. This could be caused by inaccurate diagnosis and non-causal treatment in young men. Urodynamics is helpful in the differential diagnosis and treatment of such cases. OBJECTIVES: Examination of lower urinary tract functions in young men and causal treatment of both bladder neck obstruction or impaired bladder. METHODS: In a prospective study, a group of 38 young men were treated at mean age of 42 years (range 19-50 yrs). Chronic abacterial prostatitis was treated in all cases unsuccessfully. Patients with positive urinary infection, previous surgery of the lower urinary tract and neurogenic bladder were excluded. Urodynamics confirmed a bladder neck obstruction or impaired bladder, the symptom score revealed subjective difficulties (maximum 35 points). Patients with the obstruction underwent transurethral incision of the bladder neck. Patients with impaired bladder were administrated with distigmine bromid (Ubretid) 5 mg for 1 year, every other day. All patients were re-examined one year following the treatment. RESULTS: Bladder neck obstruction occurred in 18 cases, and impaired bladder in 20 cases. Significant differences were found in relation to age (47 vs. 31 years, p < 0.01) and detrusor pressure at maximum flow (62 vs. 30 cmH2O, p < 0.01). There were no differences in peak flow rate (9 vs. 10 ml/s, p = 0.75), symptom score (19 vs. 18, p = 0.46), residual urine (45 vs. 100, p = 0.08) and maximum cystometric capacity (341 vs. 383 ml, p = 0.10). Transurethral bladder neck incision or distigmine administration improved the symptom score by 68.4% vs. 33.3%, peak flow rate 50.0% vs. 23.1% and residual urine 100% vs. 75%. CONCLUSIONS: Treatment of lower urinary tract disorders is successful in causal treatment of bladder neck obstruction and impaired bladder. (Tab. 2, Fig. 2, Ref. 12.) PMID- 11039198 TI - [Treatment of ascites with reinfusion of ascitic fluid concentrate]. AB - Ascites which is refractory to common therapeutical measures is a great problem. It deteriorates patient's life and is a sign of poor prognosis. Different methods of peritoneal fluid reinfusion belong to effective non-pharmaceutical therapeutic approaches. Main target of this study was to analyse the effectiveness and safety of peritoneal fluid reinfusion and to compare two methods of its administration (i.v. reinfusion and intraperitoneal reinfusion). During three years we have performed 97 reinfusions in 4 patients (2 women and 2 men; mean age 56 years). I.v. reinfusion was administered 68 times and intraperitoneal ewinfusion was performed 28 times. Usually we evacuated 8000 ml of ultrafiltrate. The most common complications were haemoperitoneum (6x) and short-term chills (2x). We didn't have any complications such as coagulopathy, peritonitis or circulation collapse. Intraperitoneal administration seems to be more advantageous when compared with i.v. application, because of less frequent detection of fibrin degradation products and D-dimers after the procedure and higher diuresis during the following days. (Tab. 2, Fig. 5, Ref. 13.). PMID- 11039199 TI - [Microsurgical treatment of tumors and vascular lesions in the brain stem]. AB - The authors present their experience with microsurgical replacement of brainstem lesions within the period 1989-1999. They operated on 35 patients with tumors and with KM? at age ranging from 2 to 65 years. The children suffered prevailingly from gliomas. KM was more frequent in adults. Ten adults were treated for tumors (4 gliomas, 3 haemangioblastomas, 2 primary lymphomas and 1 epidermoid). A correct surgical technique by use of microsurgical technology can replace relatively safely replace the tumor and vascular lesions even from the inside of the brainstem. PMID- 11039200 TI - [The female prostate. Update 1999]. AB - The author presents new original results from the research of the female prostate. He describes physiologic and pathologic morphology and functions of the female prostate. The mapping of female prostate diseases incidence and acceptance of the non-vestigial conception of the female prostate as a functional genito urinary organ open new possibilities in the clinical research of the female prostate and therapy of its diseases. PMID- 11039201 TI - [DNA analysis of partial and complete hydatid mole]. AB - DNA analysis enables an unambiquous recognition of partial and complete hydatiform moles and at the same time to distinguish the homozygous and heterozygous forms of KHM, the fact of which has its significance for target investigation of the course of the post-intervention period in mostly endangered patients. All investigated patients with gestational choriocarcinomas are proved to have molar pregnancy in their case histories. PMID- 11039202 TI - Alpha 2-b interferon and farmarubicin in the prophylaxis of recurrence of superficial transitional cell carcinoma of the urinary bladder. AB - OBJECTIVE: To ascertain the effect of intravesical instillation of Alpha 2-b Interferon (IFN a-2) 10 million I.U. in 50 ml physiological saline as a monotherapy and in combination with Farmarubicin (FRC) 50 mg dissolved together in 50 ml of physiological saline. These substances were administered four times during the first month after TUR-BT and then once monthly for one year either in the form of an IFN a-2 monotherapy or as an IFN a-2 and FRC combination in the therapy of recurrence of transitional cell carcinoma (TCC) of the urinary bladder after transurethral resection of the bladder tumor (TUR-BT). THEORETICAL CONSIDERATION: One of the causes of malignancy is an irreversible shift in the balance between protooncogens and tumor supressorgens. In the genetical process of the control of cell apoptosis, an important role is played by the tumor supressorgen p 53. By the means of mutation of protooncogenes, cellular oncogenes(C-MYC) are formed, inducing the proliferation of cells of the tumor and via feedback induce also the p53 mutation. By the reduction of cellular oncogenes, IFN a-2 and FRC intervene by blocking the proliferation of tumor cells. PATIENTS AND METHODS: Authors have checked and treated 33 patients (pts) with recurrent TCC. The first group of 20 pts were after TUR-BT with BCG unsuccessful intravesical therapy, and 13 pts in the second group were with recurrence of TCC, but contraindicated for BCG treatment. These 33 pts (the first and second groups) were compared with 33 pts of the third group after TUR-BT but without intravesical instillation therapy. The pts of the third group did not suffer from any other significant ailment. IFN a-2 monotherapy (10 mill. I.U./or a combination of IFN a-2 + FRC (50 mg/50 ml solution were administered for 2 hours, 1 week after TUR-BT. During the first month, instillations were done weekly, from the second to the twelfth month only once monthly. The results were evaluated for 12 to 33 months (median: 24 months). RESULTS: Group I: From 20 pts after TUR-BT + unsuccessful BCG + IFN a-2 monotherapy recurrence was registered in 4 pts (20%). Group II: Out of 13 pts after TUR-BT + IFN a-2 + FRC, recurrence was registered in 3 pts (23%). Group I + II: Recurrence in both groups was observed in 7 pts (21.2%). Group III: Out of 33 pts after TUR-BT without immuno- et chemotherapy recurrence was registered in 18 pts (54.5%). After one year of treatment, patients were checked for 24 months. The transition into an invasive tumor was observed in 4 pts (12.1%). In the comparative group of 33 pts without instillation after TUR-BT, recurrence was detected after one year in 18 pts (54.5%) and the transition into an invasive tumor was observed in 7 pts (21.2%). CONCLUSION: Intravesical instillation of BCG used to be the most frequently applied therapy following TUR-BT. The toxicity of this vaccine as well as the contraindication of this treatment in some diseases, and also the primary or secondary resistance of TCC to BCG have challenged the search for alternative possibilities of the intravesical instillation treatment. IFN a-2 monotherapy and IFN a-2 in combination with FRC are new alternative approaches in the improvement of TCC treatment. This therapy is also supported by research of molecular genetics. (Ref.18.) PMID- 11039203 TI - [Barrett's esophagus]. AB - Barrett's esophagus is a pre-malignant change in esophageal mucosa. Its relation to the reflux disease of the esophagus and the origin of adenocarcinoma (which currently has an increasing incidence) experiences its renaissance. The pathogenesis of this disease is based on the composite effect of reflux contents of hydrochloric acid and duodenal juices. In addition to endoscopy and histology, chromoendoscopy and fluorescent endoscopy are gradually starting to be put into diagnostic routine practice. The control of patients is a necessity which depends on the degree of pathohistological changes within the epithelium. The therapy of the Barrett's esophagus can be either conservative which is currently carried out by proton pump inhibitors or surgical. Ablation of epithelium in combination with a subsequent long-term antisecretory therapy appears to be a promising treatment. (Ref. 23.) PMID- 11039204 TI - [Tendon transfer of the extensor carpi radialis longus muscle in tendon rupture of the extensor pollicis muscle]. AB - The most common ruptures of all of the tendons of the hand are those of the extensor pollicis longus tendon (EPL). As to its etiology, the rupture of EPL can spontaneously occur tendosynovitis in rheumatic patients or rarely in result of conservative treatment of distal forearm fractures. During the last 18 months, 4 cases of EPL ruptures following the conservative treatment of distal forearm fractures have been diagnosed in our clinical material. All patients underwent the transfer of extensor carpi radialis longus (ECRL) tendon due to. We present our indications of the technique, and the clinical results. (Fig. 3, Ref. 8.) PMID- 11039206 TI - [Non-selective cationic current--the basis of depolarization in smooth muscle]. AB - The activation of smooth muscle muscarinic receptors leads to its contraction. Electrophysiological and biochemical methods have gradually, over the time span of roughly 30-40 past years, helped to reveal this phenomenon's underlying processes. One element of this cascade of processes is the influx of cations into the smooth muscle cell--non-selective cationic current--causing depolarisation of the cell membrane and subsequent opening of voltage-operated calcium channels. This mini-review provides the reader with up-to-date knowledge on non-selective cationic current, supplemented with information on muscarinic receptors of smooth muscle, their coupling with effectors via G proteins, as well as on muscarinic modulation of calcium and potassium channels of smooth muscle. The conclusion is dedicated to pointing out the future trends of research in this area. (Tab. 2, Fig. 2, Ref. 82.) PMID- 11039205 TI - [Primary pulmonary hypertension]. AB - Authors described a case of 26-year old patient with history of progressing dyspnea and repetitive syncopes. In history is intermittent hemoptysis and dyspnea from 1990. Diagnosis of thromboembolic disease was suspected. Clinical picture was dominated by dyspnea, central cyanosis, sinus tachycardia without pulmonary signs of hearth failure. On ecg there is right heart hypertrophy. Echocardiographic examination shows dilatation of right heart, systolic pressure in a. pulmonalis about 90 mmHg and tricuspidal regurgitation of the III. degree. Phlebothrombosis was not found. Complete hemocoagulation examination excluded a primary procoagulating hematologic disease. Pulmonary angiography did not confirm thromboembolic disease but found a high grade pulmonary hypertension--mean pulmonary arterial pressure of 93 mmHg. After complex pneumological examination, including HRCT, and other examinations the diagnosis of primary pulmonary hypertension was made. Patient is indicated to lung transplantation. (Fig. 4, Ref. 9.) PMID- 11039207 TI - [The niches and pathways of animal pathogens]. AB - Infectious diseases are not a relict of the past but a topical phenomenon determined by complex evolution of the currently existing and constantly changing microbial agents and their hosts. With regard to abundance of species within the microbial kingdom and rate of its changes and development, it is difficult to predict the role of the microbial factor in mortality of humans and animals. The study and generalization of sequential similarities of microbial virulence factors after the completion of genome sequencing of principal pathogens can play a positive role in this direction. At present, molecular-genetic methods allow us to study the phylogenetic relationships of microbes and categorize them according to new criteria. The efficient control of diseases caused by microbes requires knowledge on their physiological and ecological niche from which they penetrate, in various ways, into the host organisms and, under suitable conditions, induce mass diseases. This process has several stages and, in the recent period, it is increasingly affected by human activities. The knowledge on all participants in this process, i.e. the microbe and its niche, factors of virulence and pathways of their dissemination, requires a scientifically based surveillance. Abundance and variability is characteristic for both microbial kingdom and microbial niche. Some identification of pathogenic properties of microorganisms and factors affecting their movement from their niche to the recipients results in activation of old classical diseases (e.g. plague, cholera, tuberculosis etc.) or emerging of new, so far unknown infections diseases ("emerging inf. disease"--EID), caused for example by lentiviruses, oncoviruses, filiviruses, bartonella, borrelia etc. This has provided the basis for establishment of new medical trends and approaches, such as "Emergency medicine" or "Travel medicine", expressing their purpose by their names. The control of existing or proposed infectious diseases in the 21st century (in which majority of factors such as urbanization, environmental factors, evolution of the microbial kingdom, will contribute to the persistence or "emergence" of new diseases) will be affected by the input of new knowledge in the field of molecular biology, such as introduction of biosensors, genetic tests, microchips, new generation of DNA vaccines, enteric vaccines and antibodies produced by transgenic animal bioreactors or plants, "customized" vaccines assessed for individual genetic profiles, etc. (Tab. 5, Ref. 21.) PMID- 11039208 TI - [Trends in occupational skin diseases in the Slovak Republic]. AB - The authors present the onsets of occupational dermatoses abroad and in the Slovak Republic, organizational measures, the so-called consulting days of commissions which co-operate with medical centres of occupational hygiene. They analyse the principles of diagnosis, relevance of epicutaneous tests, criteria of professionality, claims service, judgement and compensations of admitted claims. They analyse the developmental trends of claimed occupational skin diseases and skin infections which occurred within 1973-1998 according to their individual inducers divided into 20 classes, as well as to number of claims. They review the order of inducers according to their significance: oil products, plastic materials, rubber and rubber chemicals. The 4th class includes "other chemicals" which are to be divided according to their individual chemical composition. The proportion of occupational skin diseases in relation to all occupational diseases has gradually decreased from 51.7% in 1973 to 16.3% in 1998, in absolute figures from 382 cases in 1973 decreased to 60 cases in 1998. The authors analyse the reasons of this decrease. (Tab. 4, Ref. 39.) PMID- 11039209 TI - [Primary ciliary dyskinesia--importance of early diagnosis]. AB - BACKGROUND: Primary ciliary dyskinesia (PCD) is an inherited disease characterized by specific ultrastructural defects of cilia and sperms. The impairment of mucociliary clearance (MCC) results in chronic respiratory infections and subsequently in bronchiectasis. MAIN PURPOSE: The evaluate rational decisions in early diagnosis of PCD. METHODS: Samples of nasal mucosa or tissue of tonsilla pharyngea after adenotomy were studied by transmission electron microscopy (TEM) in 47 patients aged 1-15 years, suffering from recurrent or chronic respiratory infections. RESULTS: Congenital ultrastructural ciliary defects specific for PCD--the lack of dynein arms, radial spokes defects and microtubular transposition--were observed in 13 patients. TEM investigation is an expansive, time consuming method not available in routine practice. Therefore we have evaluated a diagnostic procedure which uses available examination methods focused on the diagnoses of PCD. TEM of respiratory cilia is indicated in patients with situs viscerum solitus if chronic respiratory disease develops and after more frequent causes--asthma, cystic fibrosis, congenital anomalies of respiratory system and immunodeficiency had been excluded. CONCLUSIONS: The correct and early diagnosis is important for effective therapy in order to improve MCC. This approach can prevent the development of bronchiectasis during childhood. PMID- 11039210 TI - Lung transplantation today. AB - Lung transplantation has become an accepted surgical modality, and it is indicated in patients with a long-term benign pulmonary disease in stage when all the other therapeutic possibilities failed. Nowadays it presents a real possibility to significantly improve the quality of life. Success, (mainly in the last decade), establishing international professional centers, national coordinations, shifts transplantation towards the standard treatment procedures. The objective of the paper is to offer an overview of the international activities, trends and results in the area of lung transplantation. Authors present a review of the current situation based on their own experiences gained from the bilateral cooperation with Vienna Transplant Group. (Tab. 4, Fig. 4, Ref. 19.) PMID- 11039211 TI - The sensitivity of tussiphonography for assessing the effectiveness of treatment. AB - Our previous studies have demonstrated that tussiphonogram is suitable not only for the detection of pathological condition in the respiratory tract but also for treatment effectiveness assessment. The purpose of this study was to evaluate the possibilities of tussiphonography in detection of already little pathological changes in the airways and lungs. Therefore the changes of voluntary cough sound indexes were compared with pulmonary function tests in selected group of asthmatics before and after a pulsatile electromagnetic therapy in which the effect of therapy on pulmonary function tests was minimal. After magnetotherapy in 18 patients with increased expiratory forced lung capacity by 7.3% and increased peak inspiratory flow by 31.7% in average the voluntary cough sound intensity decreased by 37.8%, the sound duration shortened by 11% and the sound pattern showed the tendency to normalization. The improvement of mentioned cough indexes was absent in 17 patients who were treated by magnetotherapy too, but at the same time suffered from respiratory viral infection and in 22 patients treated only with climatotherapy and antiasthmatics. Changes of flow-volume loops in patients were not in the close relation to other followed indices. The correlation analysis showed a functional connection in relative differences of cough sound indices and some pulmonary function tests. The results confirmed the suitability of tussiphonography to indicate even mild pathological changes in respiratory tract. (Fig. 4, Ref. 21.) PMID- 11039212 TI - [Continuous flow ventilatory support using a multijet insufflation catheter. Physical, mathematical and clinical prerequisites and principles]. AB - The authors present theoretical principles of a new ventilatory support continuous flow ventilatory support (CFVS) with multijet insufflation catheter (MIC). Theoretical part of the presented work reasons the need of this type of ventilatory support and explains basic mathematical and physiologic principles of described mechanical ventilation method and reveals the advantages of continuous flow ventilatory support with multijet insufflation catheter in comparison with terminal eye catheter. Physical and mathematical analysis on a model of lungs in static and dynamic conditions revealed that the difference in the value of maximal inspiratory pressure is significantly higher in the system with terminal eye catheter and confirmed that the CFVS application with multijet insufflation catheter is connected with minimal risk of barotrauma by gas flow up to 20-26 l/min. The paper concludes that continuous flow ventilatory support with multijet insufflation catheter is more efficient with the possibility of significantly higher gas flow application than with terminal eye catheter and without the risk of pressure rise in the airways and without rise of breathing work. (Fig. 10, Ref. 11.) PMID- 11039213 TI - [Continuous flow ventilatory support with multijet insufflation catheter. Initial clinical experience]. AB - BACKGROUND: Clinical application of continuous flow ventilatory support with multijet insufflation catheter is not mentioned in the literature until now. Despite the use of various forms of ventilatory support, in 10-30% of patients disweaning from mechanical ventilation is unsuccessful even if they fulfil clinical and biochemical criteria. AIM: To evaluate the efficiency of a new ventilatory support continuous flow ventilatory support with multijet insufflation catheter--in clinical conditions. METHODS: Continuous flow ventilatory support with original, patented multijet insufflation catheter with nasal installation into the trachea was used in a group of 14 patients. In a subgroup of 8 patients with chronic obstructive lung disease (COLD) was this method used for development of global respiratory insufficiency due to infectious complications and in a subgroup of 6 patients it was successfully used as a ventilatory model for weaning of patients from longterm mechanical ventilation in whom other ventilatory modes for weaning were unsuccessful. RESULTS: No patient with COLD had to be intubated and 30 minutes after the start of ventilatory support with multijet insufflation catheter mean respiratory frequency decreased from 33 +/- 2.8 to 27 +/- 2.5 d/min, paCO2 from 11.9 +/- 1.7 to 10.8 +/- 1.6 kPa and paO2 increased from 5.7 to 6.8 +/- 1.3 kPa by FiO2 of 0.3. Up to 24 h after the start of ventilatory support blood gases were improved to values characteristic for partial respiratory insufficiency. Frequency of spontaneous ventilation decreased to 20 +/- 2.2, paCO2 decreased to 6.4 +/- 1.2 kPa and paO2 continually increased reaching the value of 8.9 +/- 1.4 (FiO2 = 0.3) in the 24th hour of ventilatory support. Ventilatory support lasted in average 5 days, than it could be removed. In the second group of patients continuous flow ventilatory support was used because of unsuccessful weaning following longterm mechanical ventilation. After extubation and 30 minutes after continuous flow ventilatory support start the breathing frequency decreased to 27 +/- 2.5 d/min, paCO2 showed further fall to the value of 3.9 +/- 0.9 due to hyperventilation caused evidently by continuing paO2 decrease to the value of 8.8 +/- 1.4 kPa. Only after 60 minutes following the start of ventilatory support, by equal breathing frequency, the values of blood gases increased (paO2 to 9.9 +/- 1.5 kPa, paCO2 to 5.2 +/- 1.1 kPa) and also Vt increased (0.38 +/- 0.3) which allowed to carry on with continuous flow ventilatory support, it could be interrupted after 48 hours. CONCLUSION: On the basis of the obtained results it can be stated that continuous flow ventilatory support represents an efficient ventilatory mode in patients with chronic obstructive lung disease with global respiratory insufficiency and enables to bridge the period of management e.g. of infectious complications without intubation and mechanical ventilation. As a noninvasive ventilatory regime it can be also used of patients from longterm mechanical ventilation. Application in acute respiratory for weaning (ARF, ARDS) requires further prospective studies. (Tab. 5, Fig. 4, Ref. 28.) PMID- 11039214 TI - [Endoscopic therapy of pancreatic pseudocysts]. AB - BACKGROUND: Pancreatic pseudocysts are a complication of both acute and chronic pancreatitis. Incidence in patients with acute pancreatitis is 2-50%, in patients with chronic pancreatitis 20-40%. Pseudocysts are a cause of many symptoms, e.g. nausea, vomitus, pain, biliary obstruction, bleeding and perforation. Successful treatment of pseudocysts is not only surgical and percutaneous, but also endoscopic. OBJECTIVES: The aim of this study was to answer the following questions. First, what is the clinical success rate of endoscopic drainage of pancreatic pseudocysts? Second, what are the complications? Finally, how often is endoscopic drainage a definite treatment? METHODS: The records of all patients (11) with chronic pancreatitis and endoscopic drainage of symptomatic pseudocysts hospitalized between December 1993 and April 1999 at our clinic were retrospectively studied. RESULTS: Patients (5) were followed for a mean duration of 30 months. Endoscopic drainage was definitive treatment in 80%, after transgastric drainage in 50%, after transpapillary drainage in 100% and after the use of more than one drainage procedure in 0%. The prognostic factors for longterm success of endoscopic drainage could not be evaluated, because of the small number of treated patients. CONCLUSIONS: Endoscopic treatment of pancreatic pseudocysts (endoscopic cystogastrostomy, cystoduodenostomy and transpapillary drainage) is nowadays highly effective method, technically feasible in most patients, with a relative degree of safety when performed by experienced endoscopist. (Tab. 2, Ref. 16.) PMID- 11039215 TI - [Oxygen transport values in patients with surgery performed under extracorporeal blood circulation]. AB - The aim of the work was to study the O2-transport changes to tissues in cardio surgical patients suffering from CAD and operated during extra-corporeal blood circulation (ECC). The changes of selected haematologic variables, 2,3 diphosphoglycerate (2,3-DPG) and ATP concentration, acid-base balance parameters with p50 calculation were measured in the venous blood samples taken before the operation, during the operation and on the 1st, 2nd, 3d, 5th, 7th and 10th day after the operation. From the obtained results follows, that extreme haemodilution causes significant decrease of the haematocrit value (Htc) by 35%, the value of haemoglobin (Hb) by 37% and the count of erythrocytes (Er) by 37% from the initial values. The count of reticulocytes (Ret) was increased by 52%. In the days after operation the increase in Htc values, the values of Hb and count of erythrocytes was observed, whereby the initial values were not reach even on the 10th day after the operation. The increase of the reticulocytes count by 33% prevailed to the 10th day after the operation in comparison with the initial values. 2,3-DPG concentration was increased between 3d and 10th day after the operation by 30% and ATP concentration between 5th and 10th day was increased by 23% from the initial values. Hb-O2 saturation (SpO2) and pO2 were increased already during the operation, the increase prevailed until the 7th day by 27%, pO2 until the 3d day by 39% from the initial values. Calculated values p50 did not change in the course of this study--they fluctuated in range +/- 0.04 kPa from the initial value 3.55 kPa. Supposing multifactorial character of Hb oxygenation and deoxygenation process it is possible to conclude, that the determined changes of observed parameters did not significantly influence O2 transport to tissues during ECC. (Fig. 3, Ref. 12.) PMID- 11039216 TI - [Cephalometric analysis of distance x-ray images in children with orofacial clefts]. AB - The authors with the help of cephalometric RTG analysis study the changes of facial skeleton in children with orofacial clefts and the treatment results. Cephalometric analysis was performed on distance X-rays in patients with orofacial clefts from the whole Slovak republic in the number of 178, with age of 7.8 and 9 years. According to the type of the fissure the patients were divided in four groups. The control group consisted of 46 healthy children with the same age. The method of Schwartz was used (1958). The analysis revealed some characteristic changes in patients with orofacial clefts, which document that not only local disorder of lips, alveolar process and palate develops in these patients, but the fissure defect affects also more distant parts of visceral skeleton and important growth centers. (Tab. 5, Fig. 1, Ref. 14.) PMID- 11039217 TI - [Treatment of hyperparathyroidism at the surgery department of a district hospital]. AB - Surgical treatment of primary and secondary hyperparathyreoidism in two patients in a district hospital under certain conditions, which are regular surgical procedures on the thyroid gland according to recent trends and the possibility of very quick bioptic diagnostic of the extracted tissue are presented. (Tab. 3, Ref. 14.) PMID- 11039218 TI - [Bone density and its quality]. AB - The paper presents the newest knowledge on bones quality, which becomes the main criterion for evaluation of fracture risks and osteoporosis treatment effects. It is necessary to develop new methods of measurement and quantification of bones quality, with regard to the properties of all bone structures, not solely the bone mineral density (BMD) as it was until now. From the most recent studies follows: 1. bone stiffness depends not only on BMD but also on the bone quality, 2. antiosteoporotic treatment can reduce the fracture incidence without significant rise of BMD, 3. BMD is not always an reliant marker of all treatment procedures efficiency in osteoporosis, 4. it is necessary to develop new diagnostic methods for bone quality quantification with the possibility of exact fracture risk determination, 5. in future the influence of antiosteoporotic drugs on bone quality, i.e. on all structures, not only BMD, will be studied. (Ref. 8.) PMID- 11039219 TI - [Determinants of the effectiveness of cochlear implants: I. Placement in the cochlea]. AB - The present paper introduces a registration method for the electrical field produced by a cochlear implant in the scala tympani in vitro. It was possible to determine the electrical potentials with high spatial resolution using an additional recording electrode. The results indicate that the efficacy of the stimulation by the more apical electrodes is higher than by the more basal ones. The reason is the decreasing cross-sectional area of the scala tympani, which represents a decreasing shunt between the stimulation electrodes. PMID- 11039220 TI - [An unusual case of gastroschisis combined with dysganglionosis]. AB - Authors describe a child with gastroschisis, which was associated with dysganglionosis. Therapy required an uncommon surgical treatment in connection with a long term parenteral and enteral nutrition. PMID- 11039221 TI - [Parenteral and enteral nutrition]. AB - Serious gastrointestinal surgeries are commonly connected with impossibility of natural alimentation. Surgical patient in the introduction of treatment is on various degree of malnutrition. Malnutrition affects about 40% of hospitalized patients. Because of poor symptomatology it remains in background of doctors concern whereby unfavourably influencing postoperative course. Complete nutrition support requires every surgical patient with a surgery excluding oral intake of food over 5-7 days. PMID- 11039222 TI - [Psychopathologic symptoms from the aspect of psycholinguistics]. AB - The authors stress the importance of differentiated, detailed psychopathologic theory with use of differentiated psychopathological dictionary. With practical use it can not compete with simple scales, inclusion and exclusion criteria, with operational techniques. It has only limited use in research, which must respect scientific principles. But it can bring the scientists nearer to understanding of "what" is psychosis and "what" is neurosis, and so on: so it favours "what" to "how". PMID- 11039223 TI - The matrix metalloproteinase-1 (MMP-1) expression in the human endometrium is inversely regulated by interleukin-1 alpha and sex steroids. AB - OBJECTIVE: To investigate the regulation of perimenstrual MMP-1 expression in human endometrium. DESIGN: In vitro study utilizing epithelial-stromal co cultures. SETTING: Cell Biology Unit, International Institute of Cellular and Molecular Pathology, and Departments of Pathology and Gynecology, Saint-Luc University Clinics, Louvain University Medical School, Brussels, Belgium. METHODS: Contact-dependent and contact-independent co-cultures were established and resulting MMP-1 gene and protein expression was analyzed by RNase protection assays and soluble-collagen assays. RESULTS: MMP-1 expression in endometrial fibroblasts is markedly stimulated by epithelial cell-conditioned medium. This stimulation can be prevented by antibodies directed against interleukin 1 alpha (IL-1 alpha). Ovarian steroids inhibit MMP-1 production by IL-1 alpha-stimulated fibroblasts in vitro. CONCLUSION: Taken together, our results suggest that epithelium-derived IL-1 alpha is the most important paracrine induced of MMP-1 in endometrial fibroblasts. However, IL-1 alpha-stimulated MMP-1 production in the human endometrium is effectively blocked by ovarian steroids. We believe that this mechanism responsible for the MMP-1 repression that occurs when systemic sex steroid concentrations are high and the MMP-1 production and activity during the perimenstrual phase when estrogen and progesterone concentrations are low. PMID- 11039224 TI - [Embryo transfer after 5 days of culture]. AB - OBJECTIVE: To find out whether: 1) the blastocysts transfer rises the successfulness of IVF and ET (G/ET) and 2) the 2-blastocyst transfer lowers the incidence of multiple pregnancies (IR). DESIGN: Prospective study of 46 patients involved in a prolonged 5-day cultivation of IVF and ET programme. SETTING: Centre for Assisted Reproduction, Clinic of Gynecology and Obstetrics, Teaching Hospital and Palacky University in Olomouc. METHODS: Patients with at least 5 acquired embryos were included in the group. The cultivation in media commercially produced by Scandinavian IVF Science took 5 days. RESULTS: In 91% of patients in the observed group (n = 46) morulas or blastocysts were acquired. The IVF and ET success rate after 5-day cultivation and subsequent transfer of two embryos was 37% pregnancies per a started cycle (G/cycle), 40% pregnancies per an embryotransfer (G/ET) and the implantation rate (IR) was 31%. Relatively high percentage of double pregnancies (53%) is alarming. Following parameters were assessed: age, sterility cause, number of previous IVF cycles, stimulation scheme, E2 level on the day of hCG application, number of acquired oocytes and pronuclear stages, percentage of grown blastocysts, the quality of transferred embryos and endometrium thickness on the day of transfer. The only statistically significant difference was found in E2 level on the day of hCG application. Pregnant patients had lower levels of estradiol (average value 11.8 +/- 4.8 nmol/l) compared to patients who did not become pregnant (18.6 +/- 11.9 nmol/l). CONCLUSION: In selected groups of patients who refuse multiple pregnancies only one blastocyst should be transferred. PMID- 11039225 TI - [Fetal nucleated erythrocytes in the peripheral circulation of pregnant women as a noninvasive method of fetal sex determination]. AB - OBJECTIVE: Determination of the sex of the foetus from enriched nuclear red blood cells (NRBC) circulating in maternal blood during pregnancy. METHODS: NRBC were enriched from 13-28 ml peripheral blood of 32 pregnant women using double MACS procedure. NRBCs were enriched by magnetic activated cell sorting using anti-CD71 (transferrin receptor) monoclonal antibodies. Unwanted leukocytes were depleted using monoclonal antibodies against CD14 and CD45. The sex of the foetus was analysed by using dual-colour FISH with X and Y specific probes. The experimental results obtained from the noninvasive procedure were compared to karyotype obtained from amniocentesis or chorionic villus sampling. RESULTS: In 15 out of 17 male foetuses we could identify one X and one Y signal. In another 15 pregnant women carrying female foetuses two X signals were observed. CONCLUSION: NRBC circulating in blood of pregnant women can be used as an alternative source for determination of the sex of the foetus with a risk of false negative results (2/17, 12%). The problem of false negative results can be solved by using more sophisticated methods of enrichment and preparedness of the slides for FISH analysis. PMID- 11039226 TI - [Continuous monitoring of fetal oxygen saturation (FSpO2) using intrapartum fetal pulse oximetry (IFPO) in the diagnosis of acute fetal hypoxia]. AB - OBJECTIVE: The aim of the study was to evaluate the possibility of lowering the Caesarean Section rate in patients presenting the signs of intrauterine hypoxia on CTG tracing by evaluating the foetal oxygen saturation (FSpO2) by means of intrapartum foetal pulse oximetry (IFPO). DESIGN: Open prospective study. SETTING: 1st Department of Gynecology and Obstetrics, Medical Faculty of Masaryk University, Brno. METHODS: From January 1, 1999 to December, 1999 68 patients were enrolled in the study. For the application of the IFPO sensor the patient had to meet the following criteria: patient's informed consensus, pregnancy > or = 36 weeks, regular uterine contractions, rupture of membranes, cervical dilatation of > or = 2 cm, singleton pregnancy, cephalic occiput presentation, no sings of vaginal infection, acute foetal hypoxia on CTG tracing: (baseline heart rate < 100 beats/min of different patterns. Progressive bradycardia: baseline heart rate gradually decreases between contractions (DIP II, DIP 0). Persisting bradycardia, baseline < 80 beats/min. Baseline tachycardia (> 150 beats/min) with reduced variability and/or severe variable (DIP 0) and late decelerations (DIP II). The IFPO used--Nellcor N-400. In all patients that fulfilled the above mentioned criteria during the first stage of labor the sensor was applied preferably on the posterior cheek of the foetus and the FSpO2 values were continuously monitored up to the complete dilatation. The threshold of the intrapartum foetal hypoxia (FSpO2 values) was considered < 30% for more than 10 minutes. In cases of normal FSpO2 values the delivery was conducted vaginally even if the CTG tracing continued to signalise++ intrauterine hypoxia. In case of pathologic FSpO2 values, Caesarean Section was performed. RESULTS: IFPO is an easy feasible method and in all cases the values of FSpO2 were obtained. The method has no serious side effects neither in the mother nor in the foetus. Nevertheless the presence of the sensor in the uterine cavity provokes often unpleasant feelings and limits the mother in free movements. In all suspicious CTG tracings (17) no Caesarean Sections were performed after the verification of the foetal hypoxia by means of FSpO2 evaluation. In 51 patients a pathologic CTG tracing indicating the performance of Caesarean Section was present. After FSpO2 evaluation the Caesarean Section was performed only in 11 (21.6%) patients. The remaining 40 (78.4%) delivered vaginally. Between these two groups there was statistical difference in the values of FSpO2 and postpartum cord pH. The state of newborns evaluated according to the Apgar score did not significantly differ in the two groups. CONCLUSION: These preliminary results indicate that taking in an account foetal SpO2 evaluated by IFPO in the 1st stage of labor in cases of pathologic CTG tracing (late and variable deceleration) indicating Caesarean Section, > 50% of these may be saved with identical perinatal outcome (Apgar scores, cord pH). PMID- 11039227 TI - [Does reduction in multifetal pregnancy increase the risk of poor outcome in twin pregnancy?]. AB - OBJECTIVE: To compare perinatal results for multi-foetal pregnancies where the reduction was performed with pregnancies where the reduction was not performed. SETTING: Sanatorium Pronatal, Na Dlouhe mezi 4/12, 147 00, Praha 4--Hodkovicky. METHODS: We have analyzed results of pregnancies, after delivery, for women with twin pregnancy which originated in our sanatorium, as a result of treatment with assisted reproduction methods, in the period of time from January 1st 1996 to December 31st 1998. In the group being monitored there were 122 twin pregnancies originated as a result of reduction of triple and more-foetal pregnancies. We evaluated the percentage of miscarriages, length of pregnancies, weight of the newborns and the manner of termination of the delivery. These results were compared with our control group consisting of 180 cases of twin pregnancies which were not a result of reduction. RESULTS: Analysis was performed for those mothers only where complete data were available. At a 5% level of statistical significance, it was not proved that both groups differed in average term of pregnancy or average weight of the twins. Average age of the mothers differed at 5% level of significance (average age values were 30.16 for the group with reduction and 31.73 for the group without reduction). Fisher test on 5% significance level did not ascertain any significant difference in the probability of miscarriage between the group with reduction (5.26%) and the group without reduction (12.84%). At 5% level of statistical significance, no significant difference in probability of perinatal death of the foetus or delivery of a stillborn foetus was ascertained. However, it is necessary to point out a low frequency of these phenomena in our group. The percentage of cesarean sections did not differ significantly in both groups (86.24% in the group monitored vs 87.24% in the group of twins without reduction). CONCLUSION: The analysis of both groups proves that reduction of multi-foetal pregnancies does not worsen perinatal results in comparison to pregnancies where reduction was not performed. PMID- 11039228 TI - [Management and methods of delivery in women with aortic coarctation--results of 64 pregnancies in 41 women]. AB - OBJECTIVE: To evaluate maternal morbidity and mortality of women suffering from coarctation of the aorta as well as the perinatal mortality of babies delivered by women with this anomaly of the aorta. According to our experience we recommend a suitable form of a follow up and suggest an optimal mode of delivery for these patients. DESIGN: Original paper. SETTING: 1st and 2nd Department of Gynecology and Obstetrics of the Masaryk University of Brno--Maternity Hospital, Obilni Trh 11, 602 00 Brno, Czech Republic. METHODS: Our set consists of 34 pregnant women with coarctation of the aorta. These patients were followed up during the pregnancy in the years 1964-1998 by the Centre for Cardiovascular Diseases in Pregnancy in the Maternity Hospital in Brno. Seven women who were not operated on for the coarctation of the aorta (group A) were pregnant 14 times. Twenty seven women who were operated on for the coarctation (group B) had 50 pregnancies. RESULTS: There were no maternal deaths in our set. From the 12 delivered babies of women with non operated coarctation of the aorta one child was SGA (small for gestational age). From 42 babies born by women who underwent an radical operation of the coarctation of the aorta previously we had to face one death of a newborn who was SGA as well. CONCLUSION: The radical operation of the aorta should be carried out during the first year of age if possible, between the 9th to 12th month, at best. With women who were not operated on there is a greater risk of rupture of the aneurysm of the aorta and aneurysm of the cerebral arteries in the 2nd and 3rd trimester, during the labor and in the early puerperium. We would advise therefore a through follow up during the whole pregnancy. The high BP should be lowered medicamentally. As to the mode of delivery Caesarean section is preferred. With women who were successfully operated on in their early infancy and whose BP is normal or the systolic BP does not exceed 160 mmHg the Caesarean section is not mandatory but elective. We would mostly advise a spontaneous delivery with shortening of the 2nd stage of the labor by forceps or vacuumextraction--the delivery by Caesarean section in accordance with usual obstetrical practice. PMID- 11039229 TI - [Cervix length measured by transvaginal ultrasonography in women with twin pregnancy--prospective study]. AB - OBJECTIVE: To evaluate cervical length in twin pregnancies measured by transvaginal ultrasonography. DESIGN: A prospective clinical study. SETTING: Department of Obstetrics and Gynecology, Medical Faculty of Palacky, Olomouc, Czech republic. METHODS: Clinical data of cervical length in twin pregnancies measured by transvaginal ultrasonography were summarized. RESULTS: 69% of patients delivered after 36th week of gestation, mean gestational age at the time of delivery was 35.5 week. Mean cervical length measured bi-weekly from the 16th till 34th week was 34.9, 34.6, 33.8, 33.2, 29.7, 28.7, 29.1, 28.4, 23.6, 22.8 mm. The creation of the funneling was recorded in more than 80% of patients with preterm delivery at the mean gestational age 23 week. Mean cervical length in patients delivered before 36th week was in the 23rd week of gestation 21.5 mm, while in woman delivered near term was average cervical length 33.1 mm. Genital infection in patients with preterm labours was present in 27%. CONCLUSION: Transvaginal ultrasonography seems to be a useful method in antenatal care of risk pregnancies with twins. PMID- 11039230 TI - [Development of reproductive health in women in the Czech Republic 1993-1997. III. Induced abortion]. AB - OBJECTIVE: To analyze the effect of changes in system of health and social care on reproductive health of women. In particular to analyze the changes in frequency of induced abortions as a basis for evaluation of changes in contraceptive use. DESIGN: Retrospective comparative epidemiological study. SETTING: Institute for the Care of Mother and Child, Praha 4--Podoli. METHODS: The database of the Czech Office of Health Statistics and Information was used. Besides comparison of absolute numbers of induced abortions we used international criteria "Abortion rate" and "Abortion ratio". These parameters were compared with maternal age, marital status, level of education, and number of children. RESULTS: Since the legislature on induced abortions came into effect till 1991 the annual variations of the number of induced abortions in the Czech Republic were inverse to changes in number of births due to current fertility. However, since then the number of abortions has been decreasing while the number of births was also decreasing. Using the abortion ratio as a measure, we were able to observe the decrease from 34.3 to 29.1% during 1993-1997, however the later value is still higher than in developed world. According to the selected social demographic parameters of women we have seen the highest decrease of absolute number of abortions among married women, women with two children, and women with secondary education. According to the number of abortions per 1000 women of reproductive age there has also been decrease among married women, however also among women 15-19 year and among women with university education. The smallest decrease of abortion rate has been observed among single women. CONCLUSIONS: In this part of the study we analyzed the decrease of induced abortions during 1993 1997 related to four selected social-demographic characteristics of women of reproductive age. Our results will be used for further analyses of relationship of contraception and induced abortions during the specified period. Such analysis is a subject of separate study. PMID- 11039231 TI - [Development of reproductive health in women in the Czech Republic 1993-1997. IV. Relation between contraception and induced abortion]. AB - OBJECTIVE: Overall objective is to detect changes in reproductive health of women due to changes in the system of social and health care. The particular objective of this part of the study (part IV) is to identify the changes in the relationship of decreasing number of induced abortions and increased use of contraception in 1997 compared to 1993. DESIGN: Retrospective comparative epidemiological study. SETTING: Institute for the Care of Mother and Child, Praha 4--Podoli. METHODS: Input data for the comparison of number of induced abortions and extent of contraceptive use came from the results of two previous parts of the study (parts II and III). The relationship of contraceptive use and induced abortions was analyzed with regard to possible effect of concurrent development of selected social-demographic factors. RESULTS: The highest decrease of absolute number of induced abortions, together with higher-than-average decrease of absolute number of women not using one of three effective methods of contraception, was among married women, women with two children and women with secondary education without exam. On the opposite, the highest relative decrease of women not using one of the three contraceptive methods was among women 20-24 year of age, women without children and women with university education, although such change was not reflected in relative decrease of number of induced abortions in these groups. Furthermore, their relative decrease was even smaller than the national average. Similarly, the highest relative decrease of number of induced abortions among divorced women and women 25-29 year of age was linked with less than-average decrease of number of women not using one of three effective methods of contraception. CONCLUSIONS: Different values of changing relationship of contraception and induced abortions in relation to analyzed social-demographic factors during period of observation document the influence of these factors without dispute. Due to complex influence of these factors on the relationship of contraception and induced abortions, the particular contribution of each of them is difficult to asses during simple analysis one-by-one. PMID- 11039232 TI - [Diagnosis of intrinsic urethral sphincter insufficiency in women]. AB - OBJECTIVE: Evaluation of intrinsic sphincter deficiency (ISD) according to the urethral leak point pressure in female population with stress urinary incontinence. DESIGN: Prospective clinical study. SETTING: Department of Urology, Jessenius School of Medicine Comenius University Martin, Slovak Republic. METHODS: Study population compromise of 204 females with lower urinary tract symptoms. Exclusion criteria were neurogenic bladder and unstable bladder. Valsalva leak point pressures were evaluated (VLPP). ISD was defined if VLPP below 65 cm H2O has been occurred. Pad weighing test (PWT) was positive after leakage of 2 g per hour and more. RESULTS: Stress urinary incontinence was in 134 and continence in 70 cases (control group). The first stage of incontinence was in 83, the second stage in 45 and the third stage in 6 cases of incontinence group. ISD has been occurred in 14 cases (10.5%), 8 cases with the second stage and 6 cases with the third stage. The best correlation was between VLPP and PWT, symptoms of incontinence and age of patients. All ISD cases underwent antiincontinent surgery with urethral suspension, some years ago. CONCLUSION: ISD has been risen in the second stage (6%) and the third stage (4.5%), also, in the female population with stress urinary incontinence. PMID- 11039233 TI - [Polycystic ovary syndrome as a multisystemic endocrinopathy]. AB - OBJECTIVE: To review the recent progress in the diagnosis, pathophysiology and treatment of polycystic ovary disease (PCOD). DESIGN: A review describing the progress in understanding of the dysregulation of ovarian androgen biosynthesis in polycystic ovary disease (PCOD). SETTING: The Institute for the Care of Mother and Child, Podolske nabr. 157, 147 10 Prague 4, Sanatorium Pronatal, Na Dlouhe mezi 4/12, 147 00 Praha 4-Hodkovicky. CONCLUSION: It is proposed, that although this syndrome is very heterogeneous, the final common pathway involves a dysregulation of enzymes responsible for ovarian androgen biosynthesis, possibly induced/influenced by extraovarian factors--especially insulin resistance, hyperinsulinaemie, growth factors, luteinizing hormone and their biochemical sequelae. It's probably extraovarian factors that determine endocrinological and clinical expression of the disease. PMID- 11039234 TI - [Initial experience with the Versapoint bipolar electrode in hysteroscopic tissue vaporization]. AB - OBJECTIVE: To evaluate our first experience with bipolar electrode (Verspoint, Johnson&Johnson) for transcervical surgery. DESIGN: Prospective study. SETTING: Department of Obstetrics and Gynecology, 1st medical Faculty, Charles University, Prague. METHODS: Twenty procedures with bipolar electrode in normal saline as distension fluid were performed between September and November 1999. All 20 patients underwent office diagnostic hysteroscopy with biopsy sampling which demonstrated benign histological finding. We used Olympus operative 7 mm hysteroscope, Versapoint system with bipolar electrodes (twizle, spring, ball). In 13 cases we vaporized intrauterine polyps, in 4 cases we vaporized intrauterine submucous myoma (grade I-ESH), in 1 case we dissected intrauterine septum and in 2 cases we performed intrauterine adhesiolysis. Local anesthesia paracervical block was employed in 50% of procedures and general anesthesia was employed in 50% of cases. RESULTS: The surgeon evaluated the degree of difficulty during the procedure as comfortable and easy to use in case of intrauterine polyp, intrauterine septum and intrauterine adhesions vaporisation, as mild difficulty in submucous myoma to 2 cm and moderate/severe difficulty in case of myoma vaporisation of more than 2 cm size. No complications were registered during or post procedure, we did not registered no sign of hyponatremia, no complains regarding pain or discomfort from patients. CONCLUSION: The advantage of Versapoint bipolar system for intrauterine operative hysteroscopy is the use of normal saline as distention fluid, which decreases pre and postoperative complications. We evaluated the vaporisation of pedunculate intrauterine pathologies up to 2 cm as comfortable and easy to use. Combined to local anesthesia it appears to be a useful system for office hysteroscopy and transcervical surgery. PMID- 11039235 TI - [The first pregnancy and delivery within the framework of the methadone program in the Czech Republic]. PMID- 11039236 TI - The role of tumour suppressors and viral oncoproteins in cervical carcinogenesis. AB - This article summarizes current knowledge of the cervical carcinogenesis with a special focus on the molecular mechanisms involving the interaction of cellular tumour suppressors (p53, RB, p73) with viral oncoproteins (E6, E7). The E6 induced degradation of p53 protein results in the inhibition of apoptosis, inability to repair DNA and fixation of mutations. The p53-dependent tumourigenesis is influenced by interaction not only with E6/HPV 16, 18 but also with MDM2, bcl-2 and RB protein. The polymorphism of p53 seems to contribute to malignant transformation of cervix. On contrary, there are experimental data showing that p53 may not be the only factor playing role in malignant transformation in cervical cancer. It has been generally agreed that viral oncoprotein E6 is a critical step in the onset of malignant transformation of cervix. There is a vast number of experimental and clinical studies confirming the validity of E6 induced cervical cancer including alteration of the genotype and phenotypic characteristics of the transforming cells. The modern tools of molecular biology offer an exact diagnosis as well as relevant targets for gene therapy of the cervical cancer. PMID- 11039237 TI - [Recommendations for prescribing combined oral contraceptives]. PMID- 11039238 TI - [In Process Citation] PMID- 11039240 TI - [In Process Citation] PMID- 11039239 TI - [On the article by P. Bartos: "Congenital vaginal aplasia: laparoscopic reconstruction of the neovagina by the Vecchietti method in the Mayer-von Rokitansky-Kuester-Hauser syndrome"]. PMID- 11039241 TI - [Hormonal regulation of gene transcription--specificity and regulation of the hormonal response and results of its dysfunction]. AB - Nuclear hormone receptors act as ligand activated transcription factors. Hormonal response in vivo is, however, modulated at various levels beyond the mere activation of hormone receptors by their respective ligands. There is a great variability of particular arrangements of HRE and receptors of the TR/RAR subfamily use extensive dimerization to produce a particular dimer with optimal affinity for HRE in question. Hormonal response is mediated by complex hormone responsive units (HRU), which integrate HRE together with other regulatory sequences. Some hormone responsive genes may themselves code transcription factors or other regulatory proteins, underlying delayed primary and secondary responses. In addition to transcription activation, nuclear receptors enter repressive interactions with unrelated transcription factors like AP-1 or NF kappa B. Genes for coding nuclear receptors are subjected to homologous downregulation. Functioning of nuclear receptors might be modulated by phosphorylation, which can underly ligand-independent activation. Nuclear receptors are mutated and/or disregulated in various pathologies, like hormone resistance syndromes of cancer. PMID- 11039242 TI - [Role of the kidney in long-term regulation of blood pressure and the development of hypertension]. AB - Evidence about the crucial role of the kidney in the development and maintenance of the "essential" hypertension (i.e. hypertension which is accompanied with the absence of any pathological change in any of body organs) and in the regulation of "normal" blood pressure level (BP) has been accumulated. Blood pressure is expressed as a product of cardiac output (CO) and total peripheral resistance (TPR). TPR almost entirely depends on the volume of the extracellular fluid (ECFV): With increasing volume BP rises and vice versa. ECFV--due to intensive maintenance of the osmolality--almost entirely depends on the total amount of sodium in the organism. This amount is not determined by the intake of salt, which is in every civilized population always higher than necessary. Sodium balance is therefore critically determined by the output of sodium, which is carried out almost entirely by the kidney. The output depends on the quantity of the glomerular filtration and on the tubular reabsorption. Under normal circumstances, the increased sodium intake is accompanied by an increased excretion via the mechanism called "pressure natriuresis". It is based on the prompt increase of sodium excretion after an increase of BP, resulting from the increased sodium intake. Mechanism of such elevated excretion is not clear; lot of evidence has been accumulated for the existence of a humoral principle produced within the kidney. Such assumption is supported by experiments in which a kidney is transplanted from a hypertonic donor to a normotensive recipient: hypertension in the recipient develops. Similarly, kidney grafting from a normotensive animal corrects the hypertension in an originally hypertensive recipient. Important role of the renin-angiotensin (RAS) and nitric oxide (NO) systems is often stressed in this context. If kidney is unable to excrete the ingested amount of sodium at the normal BP, blood pressure must rise and the shift of the pressure-natriuresis curve to the right is the necessary consequence. If these conditions are long lasting, hypertension develops soon and becomes "fixed" by rebuilding the resistance arteries architecture. PMID- 11039243 TI - [Lower urinary tract function and its disorders]. AB - The lower urinary tract provides two modes of operation--storage and elimination of urine. The normal function results in the coordination of contraction and relaxation of muscles of the urinary bladder and urethral sphincters. Disorders of these activities or their interaction lead to the development of lower urinary tract dysfunctions. The nervous system plays an essential role in the regulation of the functions. The control of micturition is coordinated by several regious of the central nervous system. Afferents and efferents of the peripheral nervous system carry signals from and to the lower urinary tract. The reflex circuitry controlling micturition consists of five components: spinal efferent neurons, peripheral efferent neurons, primary afferent neurons, spinal interneurons and neurons in the brain. Preganglionic neurons located in the sacral parasympathetic nucleus and lumbar sympathetic nucleus excite the peripheral efferent neurons innervating smooth muscles of the urinary bladder and urethra. Motoneurons of sacral Onuf's nucleus excite the striated muscle of the external urethral sphincter. Myelinated and unmyelinated afferent axons transmit information from the lower urinary tract to the lumbosacral spinal cord. Three receptor types of the lower urinary tract are present: tension receptors, volume receptors and "silent receptors", which become nociceptors following the sensitization. Afferent pathways terminate on spinal interneurons. Spinal interneurons relay information to the brain or to other regions of the spinal cord. Because micturition reflexes are mediated by disynaptic or polysynaptic pathways, interneuronal mechanisms are of crucial importance in the regulation of lower urinary tract. Central pathways involved in micturition reflexes are located in spinal and supraspinal areas. Micturition reflexes can be modulated at the level of the spinal cord by viscero--bladder and somato--bladder reflexes. Supraspinal areas have a more complicated organization: critical component of the micturition reflex is the pontine micturition center and the periaqueductal gray. Inhibitory and excitatory areas in the pontomedullary and hypothalamic systems and the brain play an important role in the regulation of micturition reflexes. PMID- 11039244 TI - [Pulmonary surfactant in the respiratory tract]. AB - Pulmonary surfactant is a surface active material able to reduce surface tension at the alveolar and bronchiolar air-liquid interface. It plays an important role in conductive as well in peripheral airways and that is why surfactant dysfunction is thought to contribute to pathomechanisms of pulmonary diseases with bronchial obstruction. Airway surfactant originates mainly from alveoli and some components (specific proteins SP-A, SP-B and SP-D) are synthetized and secreted also by non-ciliated cells of the airway mucosa. In comparison to alveolar surfactant the bronchiol one has the same composition, with lower amount of surfactant proteins. Surfactant has following functions in airways: stabilisation of small airways, role in mucociliary and non-ciliary transport, immunomodulation, barrier function and antiedematous function. Ability of pulmonary surfactant to influence wall thickness and diameter of airways indicates possible role of its quantitative and/or qualitative changes in the pathogenesis of asthma, chronic obstructive pulmonary disease and others. PMID- 11039245 TI - [Clinical cardiology and physiology]. AB - Recent concepts of cardiovascular pharmacotherapy are now mainly based on results of multicentre prospective studies performed in several countries worldwide. The main disadvantage of such studies is the heterogeneity of patient-groups compared, caused by ethnic differences, variability of criteria and diverse concomitant medication. To assert internal validity of data compared, substantial section of patients (80-99%) is often excluded before evaluation. Thus, there is always a lack of external validity--the data cannot be generalised and have only a limited predictive value for management of other groups or individual patients. Consequently, we still need reliable preclinical data from animal and human studies under well defined and uniform conditions. The importance of experimental and clinical physiology is demonstrated by well-established hemodynamic rules and pharmacodynamic correlations. Clinical physiology remains an indispensable foundation of clinical medicine which cannot be replaced by formal statistics. PMID- 11039246 TI - Immunoglobulin D multiple myeloma. AB - BACKGROUND: Immunoglobulin D (IgD) multiple myeloma (MM) is rare, accounting for less than 2% of all patients with MM in Western countries. In Taiwan, the frequency and clinicopathologic features of IgD MM have not yet been reported. METHODS: The clinicopathologic features and treatment outcome of patients with IgD MM diagnosed between January 1, 1982, and December 31, 1998, in Chang Gung Memorial Hospital were retrospectively reviewed. Nineteen patients with IgD MM were diagnosed. Of those patients, the medical records of 16 were available for review. RESULTS: Most of the patients were male (11/16) with a median age of 59 years. The most common presenting features included bone pain (56%), gastrointestinal discomfort (38%), general malaise (38%), and body weight loss (25%). The majority of patients had cytopenia (88%), renal function impairment (75%), hypercalcemia (63%), hyperuricemia (69%), elevated beta 2-microglobulin levels (86%), and Bence Jones proteinuria (92%). Serum protein electrophoresis showed an M-peak in 9 cases (9/12), whereas immunoelectrophoresis or immunofixation identified an IgD monoclonal gammopathy in all 16 patients. All of the M-proteins were of a lambda-light chain type. The IgD level ranged between 584 and 129,000 IU/ml (normal, < 100 IU/ml). All the patients had stage III disease except one (stage I). Four patients were still alive at the time of this analysis. The median survival time was 12 months. Infection was the leading cause of death (50%). CONCLUSION: The present series showed that IgD MM had aggressive clinical features, male predominance, a high frequency of renal function impairment, high incidence of M-protein undetected by serum protein electrophoresis, a predilection for lambda-light chains, and a short period of survival. PMID- 11039247 TI - Change of core conflicts of schizophrenic patients who received brief psychodynamic psychotherapy: a pilot study in Taiwan. AB - BACKGROUND: This study examined the change of core conflicts of schizophrenic patients who received brief psychodynamic psychotherapy (BPP). METHODS: The change of core conflicts and relationship patterns over dynamic psychotherapy sessions were assessed by the core conflictual relationship theme (CCRT) method. Patients were enrolled either from the psychiatric outpatient department or day hospital in a medical center. Transcripts of psychotherapy sessions were used for CCRT analysis. In each extracted transcript, 3 components, i.e., wishes, responses from others and responses of self, were identified and were used to formulate the CCRT. CCRT pervasiveness was calculated to compare the change of each component from early to late sessions. RESULTS: The main other person most frequently mentioned was the family member. The changes of positive and negative responses of self were found to be statistically significant. Wishes were not so pervasive as those of non-psychotic patients. A high drop-out rate characterized the schizophrenic patients. CONCLUSION: Self-understanding and self-control may well explain the significant change in responses of self, even in patients with essentially poor ego strength like schizophrenics. PMID- 11039248 TI - DNA polymorphism of Mycobacterium abscessus analyzed by infrequent-restriction site polymerase chain reaction. AB - BACKGROUND: Mycobacterium abscessus is an important pathogen that has been increasingly associated with many clinical and nosocomial infections. A reliable molecular typing scheme is essential for the epidemiological study of this rapidly growing mycobacterium. Pulsed-field gel electrophoresis (PFGE), considered to be the gold standard among molecular typing methods, has failed to provide satisfactory results in the molecular typing of this bacterium. A newly developed molecular typing method, infrequent-restriction-site polymerase chain reaction (IRS-PCR), was examined in this study to determine its suitability for fingerprinting M. abscessus isolates. METHODS: Eight clinical isolates of M. abscessus and two reference strains (M. abscessus ATCC 19977 and M. chelonae ATCC 35749) were studied by DNA macrorestriction analysis with XbaI resolved by PFGE, and IRS-PCR assay with adaptors designed for XbaI and HhaI restriction sites. RESULTS: By PFGE, different banding patterns were found in two clinical isolates of M. abscessus; the other isolates yielded only broken DNA and could not be assessed. By IRS-PCR, unique patterns were noted for the 10 isolates; the 10 appeared to be genetically different. CONCLUSION: IRS-PCR may be an efficient substitute for PFGE in analyzing the DNA polymorphism and epidemiology of M. abscessus. PMID- 11039249 TI - Gender prevalence in twin-twin transfusion syndrome. AB - BACKGROUND: This study aimed to determine the gender prevalence of fetuses complicated with twin-twin transfusion syndrome (TTTS). METHODS: All cases of TTTS corresponded with the following major criteria: a single placenta, monochorion, the same gender, and a combination of polyhydramnios oligohydramnios. At least one of three minor criteria were required for the establishment of TTTS, including a stuck twin, a birth weight discordance exceeding 20%, and hemoglobin difference > 5 g/dl. RESULTS: Fifty-six twin pregnancies met the above criteria, of which 33 (58.9%) twin pairs were female. The female tendency existed, but there was a non-significant difference. Mean gestational age at diagnosis was 20.2 +/- 3.2 weeks. The birth weight discordance exceeding 20% was present in 50 of 56 (89.3%), and mean growth discordance was 32% +/- 8%. A stuck twin was noted in 37 of 56 cases (66.1%). The mortality of fetuses or neonates was 34.8% (39/112), including 8 (7.1%) fetal deaths and 31 (27.6%) neonatal deaths. There were no differences in maternal age, parity, or gestational age of delivery between male and female pregnancies. CONCLUSION: Although the female preponderance did not reach statistical significance, the female tendency might still exist after a larger series analysis. The female tendency may be the result of the gender difference in monochorionic twins. The gender difference could provide research implications and a diagnostic warning for clinicians in monochorionic twin pregnancies before the presence of TTTS. PMID- 11039250 TI - The effects of glurenorm on plasma glucose and lipids in patients with type 2 diabetes mellitus. AB - BACKGROUND: The effects of sulfonylureas on plasma glucose, lipids, and macrovascular complications are of interest. This study was designed to investigate the effects of glurenorm on plasma glucose and lipids in patients with type 2 diabetes mellitus. METHODS: Nineteen patients, 15 men and 4 women, with an age range of 38-69 years, and with type 2 diabetes mellitus, were studied. Plasma glucose, glycated hemoglobin, and lipids were compared before and 3 months after glurenorm treatment. RESULTS: Fasting and postprandial plasma glucose, and HbA1c significantly improved after 3 months of glurenorm treatment. The mean (+/- SD) triglyceride level of 10 patients with mild to moderate hypertriglyceridemia decreased from 279 +/- 66 to 219 +/- 100 mg/dl (p = 0.054). The total cholesterol, low-density lipoprotein cholesterol (LDL-C), and high density lipoprotein cholesterol (HDL-C) of 14 hypercholesterolemic patients did not change significantly. Their mean body weight increased significantly from 65.7 +/- 9.6 to 67.2 +/- 9.9 kg (p = 0.002). CONCLUSION: Glurenorm was effective for glycemic control but caused weight gain in type 2 diabetic patients. Triglycerides in hypertriglyceridemic patients, and total cholesterol, LDL-C, and HDL-C in hypercholesterolemic patients did not improve after glurenorm treatment. PMID- 11039251 TI - Dopamine deficiency in rubral tremor caused by midbrain hemangioma: case report. AB - To elucidate the nigrostriatal involvement in rubral tremor, we studied single photon emission computed tomography (SPECT) imaging with [2-[[2-[[[3-(4 chlorophenyl)-8-methyl-8-azabicyclo[3.2.1]oct-2- yl]methyl](2 mercaptoethyl)amino]ethyl]amino]ethanethiolato(3-)- N2,N2',S2,S2']oxo-[1R-(exo exo)]-[99mTc]technetium ([99mTc]TRODAT-1) in a 70-year-old woman with a midbrain hemangioma. She had developed a slow tremor in her right arm and leg after an episode of hemorrhage at the age of 28. The tremor was 3 to 5 Hz in frequency at rest, which was enhanced by outstretching the limb and action. There was no rigidity or bradykinesia. Neurological examination also revealed mild palsy of the left oculomotor nerve. The magnetic resonance imaging (MRI) of the brain showed a small hemangioma in the left midbrain localized mainly in the substantia nigra. The [99mTc]TRODAT-1 SPECT imaging revealed significantly reduced [99mTc]TRODAT-1 uptake in the left caudate and putamen, but it was only mildly reduced in the right striatum. This reduction in uptake was even more severe than that of patients with Parkinson's disease, and indicated that the dopamine function was markedly impaired in the left nigrostriatal system. The tremor had not progressed over the years, and she responded moderately to treatment with levodopa. We concluded that the rubral tremor in the right extremities was probably caused by a dopamine deficiency in the left nigrostriatal system. This suggests that a dopamine deficiency secondary to the midbrain hemangioma might have contributed to the development of the rubral tremor in this patient. PMID- 11039252 TI - Fatal and non-fatal chromobacterial septicemia: report of two cases. AB - Chromobacterium violaceum is frequently found in soil and water in tropical and subtropical regions. The organism rarely causes infection in humans, but is associated with a high mortality rate when it occurs. Septicemia associated with abscess in multiple organs such as the liver, skin, lungs, spleen, lymph nodes, and brain has been reported. We report on 2 patients with systemic infections with Chromobacterium violaceum. One presented with a fulminant course with multiple organ involvement and died 78 days later. The other presented with a milder course and survived after antibiotic therapy. In conclusion, infection with Chromobacterium violaceum is rare but its course is usually fulminant with high mortality especially in patients with sepsis and multiple organ involvement. We hope this report will provide additional information to physicians in the treatment of this disease. PMID- 11039253 TI - Lingual osteoma: case report. AB - Lingual osteoma, a rare clinical entity, has been found mainly in the posterior region of tongue. It mostly affects women in their third and fourth decades of life and occurs less frequently in men. We report an unusual case of a 42-year old male patient who developed a lingual osteoma near the foramen cecum. The patient underwent excision of the tumor mass under local anesthesia and had an uneventful postoperative course. Symptoms of the mass effect were noted to resolve after surgical intervention. Histologically, mature lamellar bone with haversian systems was seen. The pathogenesis of this rare tumor is a controversial problem and its nomenclature also remains an issue of debate. Discussion of the controversial pathogenesis of lingual osteoma and a review of the literature on its clinical characteristics are included in this report. PMID- 11039254 TI - Malignant phyllodes tumor of the breast metastasizing to the pancreas: case report. AB - Phyllodes tumor of the breast, or cystosarcoma phyllodes, is an unusual breast tumor. It is usually considered a benign lesion but may have malignant potential. Only a small proportion of malignant phyllodes tumors will metastasize. A phyllodes tumor of the breast metastasizing to the pancreas is rare. This 39-year old female patient initially presented to us with a 5 x 4 x 2 cm tumor of a right breast. After excision of the tumor, she received another wise excision for a local recurrence of the tumor. Three years had passed when she started having hematemesis and tarry stools intermittently over a period of about one month. Clinical evaluation disclosed a huge mass with an ulcerated and bloody base over the second portion of the duodenum. A pancreatoduodenectomy was then performed. During the postoperative recovery period, a rapidly enlarging tumor of the right breast was noted. She subsequently underwent a total mastectomy of the right breast. Both the duodenal tumor and the breast tumor were found to be malignant phyllodes tumors. The rarity of this kind of patient and presentation is discussed. PMID- 11039255 TI - [beta 2-microglobulin in hemodialysis: an adequacy factor]. PMID- 11039256 TI - [Central agents in arterial hypertension: back to the future. PMID- 11039257 TI - [Hyperfiltration nephropathy]. PMID- 11039258 TI - [Why do ambulatory hemodialysis patients go to hospital emergency services? AB - An important number of Hospital admissions (HA) occurs through Hospital Emergency Departments (HED). This is a indicator of quality and have to be lower than 50%. However there are almost no data available on the causes of emergency consultation by outpatient hemodialysis patients (HD). For this reason, we prospectively examined a population of 83 outpatient HD patients dialyzed in a peripheral unit under the surveillance of a University Hospital. OBJECTIVES: 1) To know the diagnosis of HED and days of hospitalization for which HD patients came to the HED in 1998. 2) To know the possible risk factors associated with the patients with frequent assistance in HED. 3) To compare the number and causes of emergency consultation in 1998 with a group of patients treated in the same Unit in 1991 (n = 39). RESULTS: The percentage of patients who used the HED in 1998 was 66.3% (55/83). The total number of emergency episodes in 1998 was 118 (mean of 55 patients 2.27 +/- 1.51). Fifty one percent of the emergency episodes were due to patients initiative. The 4 more frequent diagnoses of HED in 1998 were infectious, 19.5% (23/118); traumatologic emergencies, 15.3% (18/118); digestive disease 15.3% (18/118); relationed problem vascular access, 11.9% (14/118). Thirty percent (36/118) of the emergency consultations needed HA leading to a mean hospitalitation of 10.2 +/- 9.3 days. The infectious disease were the highest percentage of HA (36.1%) and the longest days of hospitalitation (12.7 +/ 11.2 days). The risk factors for repeated emergency consultation (more than 3 times) were: age (68.9 vs 61.4), lower hematocrit (31.6 vs 34.4%), lower hemoglobin (10.2 vs 11), high EPO dose (166.3 vs 109.7 unit/kg/week) and lower Kt/V (0.99 vs 1.11). If we compare these results with 1991 the percentage that used the HED was similar 66.2% (pNS); the number of emergency episodes was higher (mean 2.99 +/- 1.96) than 1998 (p < 0.006) and there are a significant differences in the diagnoses of HED between 1998 and 1991: acute pulmonary edema 1.7 vs 11.2% (p < 0.003); hiperkalemia 0.8 vs 7.9% (p < 0.009); gastrointestinal disease 15.3% vs 4.5% (p < 0.008) and infectious 19.5% vs 7.9% (p < 0.01). In conclusion our study provides data previously not available on the epidemiology of Emergency Consultation by outpatient HD patients treated in the same peripheral unit. The data obtained albeit limited because of the number provide information of potential protocol usefulness for the possible reduction in the frequency of Hospital Emergency Consultations by outpatient HD patients. PMID- 11039259 TI - [Evolution of the aluminum concentration in the final dialysis solution: Multicenter study in Spanish dialysis centers]. AB - Aluminium contaminated dialysate is the most dangerous source of aluminium for dialysis patients. The aim of this study was to assess the aluminium content in the dialysis fluid in all the Spanish dialysis centres in 1999 and to compare the results with those obtained in previous studies. For this purpose, all the 275 Spanish centres were invited to participate, we measured the concentration of aluminium in the dialysis fluids in all of them. Aluminium was measured by atomic absorption spectrometry. Since 1988 our laboratory has participated in a external quality assessment scheme for aluminium measurement (University of Surrey) having a good performance (fig. 1). The aluminium concentration in the dialysis fluids were compared with the results obtained in other 2 cross sectional studies performed in 1990 and 1994 following the same methodology. The participating centres were 242 out of 275 (88%). The percentage of centres with a concentration of aluminium in the dialysis fluid lower than 2 micrograms/l has increased throughout the period of study (45% in 1990, 69.8% in 1994 and 81.8% in 1999, fig. 2). One important finding of the new study was the increment in the percentage of centres having undetectable aluminium (< 1 microgram/L) (22.9% in 1990, 41.2% in 1994 and 66.9% in 1999, fig. 3). The safety threshold of 1 microgram/L should be the goal for all the dialysis centres. By contrast, the percentage of centres with aluminium concentration greater than 10 micrograms/L (the old safety threshold to avoid aluminium exposure established by the European Union in 1986) did not show a relevant decrease from 1994 to 1999 (from 5.1% to 4.1% respectively). Taking into account the aluminium content, the quality of the dialysis fluid has improved during the last 10 years, although there is still a non negligible percentage of centres (4.1%) with high aluminium concentration in the dialysis fluid (> 10 micrograms/L). PMID- 11039260 TI - [A comparison of phosphorus-chelating effect of calcium carbonate versus calcium acetate before dialysis]. AB - INTRODUCTION: The hyperphosphatemia, hypocalcemia and low calcitriol levels are pathogenic factors for secondary hyperparathyroidism in chronic renal failure. The phosphorus control is essential to prevent secondary hyperparathyroidism. There are not comparatives studies to test the efficacy of control of phosphorus binders in predialysis patients. AIM: To compare the efficacy of calcium carbonate vs calcium acetate as phosphate binder in predialysis patients. MATERIAL AND METHODS: The present study includes 28 patients with chronic renal failure (mean clearance of creatinine 21 ml/min). Patients were separated into two groups: Group 1: (n = 14) received calcium carbonate 2,500 mg/day (1,000 mg of calcium); Group 2: (n = 14) receives calcium acetate 1,000 mg (254 mg of calcium). Calcium and phosphorus were determined every 4 months; i-PTH, alkaline phosphatase and clearance of creatinine were determined every six months. RESULTS: Both groups were comparable regarding age, renal function, calcium, phosphorus, alkaline phosphatase and i-PTH on basal situation and the end of study were not different. The serum calcium increased, not significantly, in the calcium carbonate group (group 1) [from 9.2 to 9.8 mg/dl (p = 0.05)], however it was not modified in the calcium acetate group (group 2). The serum phosphorus decreased significantly (p < 0.05) in both groups, independently of the calcium levels. Alkaline phosphatase and i-PTH not was modified during the study period. CONCLUSIONS: 1) Both calcium carbonate and calcium acetate are similarly effective as phosphate binder. 2) The carbonate group required four fold greater doses of calcium that acetate group. 3) The calcium acetate has less hypercalcemic effect than calcium carbonate. PMID- 11039261 TI - [Influencing factors in the control of phosphorus in peritoneal dialysis. Therapeutic options]. AB - Impaired phosphate excretion resulting in hyperphosphatemia is one of the earliest consequences of chronic renal failure. To control serum phosphate levels, we can use the following therapies: 1) Restriction of dietary phosphate (but on CAPD, obligatory protein losses via peritoneal fluid makes impractical any reduction of phosphate diet. 2) Reduction of phosphate absorption, using phosphate binders. 3) Peritoneal phosphate removal. OBJECTIVE: 1) To evaluate the factors affecting peritoneal phosphate removal such as plasma phosphate, peritoneal membrane transport type, peritoneal dialysis modality prescription (CAPD or APD) and daily dialysate volume. 2) To test the best calcium concentration in the peritoneal dialysis fluid (5, 6 or 7 mg/dl) in order to permit the use of calcium carbonate or acetate without the risk of hypercalcemia or hyperparathyroidism. METHOD: Phosphate was measured in seventy 24-hour dialysate collections, 33 from patients on CAPD and 37 from patients on APD. 24 hour peritoneal phosphate removal (mg/24 hours) and weekly peritoneal phosphate clearance was calculated (L/week). The peritoneal membrane was studied by the peritoneal equilibrium test (PET), using a 2.27% glucose. We calculated also the peritoneal calcium balance in 25 daily peritoneal fluid collections from patients using a calcium dialysate concentration of 5, 6 or 7 mg/dl each one. IPTH levels and doses of vitamin D were compared at 6 months in patients using a calcium concentration of 5, 6 or 7 mg/dl from the beginning of peritoneal dialysis (5 patients of each calcium dialysate concentration). RESULTS: Weekly peritoneal phosphate clearance (WPC) were higher or APD than on CAPD (51 +/- 21 vs 41 +/- 14, p < 0.005). Daily dialysate volume was also higher on APD (14 +/- 4 vs 7.8 +/ 1.8 L/day, p < 0.001). WPC was higher on APD when a mild-day exchange was done (61 +/- 23 vs 45 +/- 15, p < 0.005), instead an equal total daily volume of the dialysate. Peritoneal calcium balance was significantly more negative in patients using a calcium in the dialysis fluid of 5 than 6 or 7 mg/dl (-125 +/- 7 vs -18 +/- 41 vs -11 +/- 49, p < 0.001). At 6 months, patients using a calcium fluid concentration of 5 mg/dl increased iPTH levels (from 160 +/- 101 to 332 +/- 153, p < 0.001) and vitamin D needs (from 0 to 1.87 +/- 0.37 mcg/week, p < 0.001). In summary, peritoneal phosphate clearance depends on plasma phosphate levels, daily volume of dialysate prescribed and peritoneal membrane transport characteristics. It can be improved by increasing the total peritoneal fluid. On APD, a mild-day exchange may improve phosphate clearance, without total volume increase. The risk of secondary hyperparathyroidism can be decreased with a calcium fluid concentration of 6 mg/dl, which was shown to be better than 5 mg/dl when calcium phosphate binders are not correctly taken. PMID- 11039262 TI - [Study of renal osteodystrophy by bone biopsy. Age as an independent factor. Diagnostic value of bone remodeling markers]. AB - The spectrum of bone disease in uremic patients on hemodialysis has changed in the last years. Undecalcified bone biopsy with histomorphometric measurements and tetracycline labelling remains the gold standard for diagnosis of the different forms of renal osteodystrophy. But because of its invasive nature and complicated laboratory processing a number of non-invasive biochemical parameters have been proposed. The aim of our study was to determine the prevalence of the different forms of renal osteodystrophy in our patients in hemodialysis. Moreover we analyse the correlation between several biochemical parameters and the histological findings and evaluate their diagnostic and predictive value. Transiliac bone biopsies were performed in seventy three uremic patients (31 males) on chronic hemodialysis and static and dynamic parameters were measured. Serum levels of intact parathyroid hormone (iPTH), osteocalcin (OC), total alkaline phosphatase (FAT) and bone alkaline phosphatase (FAO) were determined. High-bone remodelling (50 pts, 68.5%) predominates over low-bone remodelling (23 pts, 31.5%). The distribution of the different types of bone disease was: Mild hyperparathyroidism 8 pts, Osteitis fibrosa 37 pts, Mixed lesions 5 pts, Adynamic bone disease 21 pts and Osteomalacia 2 pts. Six of our 73 patients were diabetics and they had adynamic bone disease (4 pts), osteomalacia (1 pt) and osteitis fibrosa (1 pt). Patients older than 50 years presented lower cellular activity (osteoblast surface, ObS/BS) and lower bone formation rate (BFR/BS). iPTH showed different correlation with these parameters of bone formation in patients above and below 50 years old suggesting that older patients need higher levels of PTH to obtain a determined level of bone formation. iPTH, OC, FAT and FAO correlated with the majority of histomorphometric indices of bone formation and resorption, though the best correlations were those with iPTH. The diagnostic and predictive value of these bone markers is better with high-bone remodelling. Serum levels of FAT > 300 U/l, OC > 150 ng/ml, FAO > 40 ng/ml and iPTH > 200 pg/ml showed a positive predictive value of 1 (with a specificity of 1, but sensibility below 0.78 except for iPTH that is 0.95) in the diagnosis of high-bone remodelling. After an analysis with ROC curves the cut-off value to differentiate high from low-bone remodelling was obtained. iPTH level > 200 pg/ml combined with one of the other markers (FAT > 150 U/l, FAO > 30 ng/ml or OC > 100 ng/ml) are predictive of high-bone remodelling, while values below those figures are predictive of low-bone remodelling. PMID- 11039263 TI - [Acute pyelonephritis. Study of 153 cases]. PMID- 11039264 TI - [Diagnostic role of magnetic resonance in the renal involvement in nocturnal paroxysmal hemoglobinuria]. AB - Paroxysmal nocturnal haemoglobinuria or Machiafava-Micheli disease is an acquired clonal stem cell disorder characterized by defective haematopoiesis, which results in an increased sensitivity of the erythrocytes to complement-mediated intravascular haemolysis. Renal damage is rare but it can lead to chronic renal failure. Micro-infarctions due to repeated episodes of microvascular thrombosis and cortical haemosiderosis are thought to be the main contributors to the development of chronic renal failure in paroxysmal nocturnal haemoglobinuria. Magnetic resonance imaging provides characteristic images of the kidney. We describe a patient with cortical haemosiderosis and show the magnetic resonance features. PMID- 11039265 TI - [Extracapillary IgA nephropathy associated with infection with hepatitis C virus and hepatic cirrhosis]. AB - We describe a 36 year old man who was admitted to the hospital with dyspnea, edema of the lower limbs, arterial hypertension and oliguric renal failure. He had microhematuria and nephrotic range proteinuria, immunological tests were normal or negative. Renal biopsy revealed global (55%) or segmental glomeruloesclerosis, remaining glomeruli showed extracapillary proliferation (25%). Immunofluorescence study disclosed IgA mesangial deposits. He was also diagnosed as having liver cirrhosis with positive serology against hepatitis C virus. He was treated with dialysis, antihypertensive drugs and steroids with improvement of the renal function. However, ten months later maintenance hemodialysis became necessary. We emphasize two points: first IgA glomerulonephritis is rarely associated with hepatitis C infection, and second crescentic IgA nephropathy has been infrequently reported in liver cirrhosis. PMID- 11039266 TI - [Progressive systemic sclerosis associated with anti-myeloperoxidase ANCA vasculitis with renal and cutaneous involvement]. AB - Sclerodema renal crisis is the usual form of presentation of renal disease in systemic sclerosis. We report a woman who at age 63 was given a diagnosis of scleroderma with Raynaud's phenomenon and cutaneous, oesophageal and lung involvement but no evidence of renal disease and no treatment with D penicillamine. Two years later she developed progressive renal failure, nephrotic range proteinuria, haematuria and the presence of serum MPO-ANCA; she was normotensive. Renal biopsy revealed extracapillary and necrotizing glomerulonephritis and skin biopsy showed leucocytoclastic vasculitis. This clinical picture was compatible with necrotizing vasculitis of the microscopic polyarterits type. After treatment with pulse steroids followed by oral steroids and monthly intravenous cyclophosphamide her renal function stabilised and the serum MPO-ANCA disappeared. PMID- 11039267 TI - [Rhodococcus equi brain abscess in a patient in hemodialysis]. PMID- 11039268 TI - [Peritoneal sclerosis after recurrent Klebsiella pneumoniae peritonitis]. PMID- 11039269 TI - [Campylobacter fetus fetus: an infection in immunosuppressed patients unknown in hemodialysis]. PMID- 11039270 TI - [Can bite force be predicted from the shape of the face?]. AB - The relationships between maximum bite force (MBF) and facial architecture have been tested in a sample of 118 dental students. The strongest correlations were seen with the variables representative of posterior face height and facial divergence. No correlation could be seen between MBF and incisor's position. MBF allowed only partial discrimination of the individuals in different subgroups constituted by cluster analysis. MBF is to consider as a representation of the individual aptitude to develop more or less powerful forces. It summarises only part of the functional information influencing facial form. PMID- 11039271 TI - [Mechanics of palatal expansion in the occlusal plane]. AB - The mechanics of a transpalatal arch in the occlusal plane (symmetrical V-bends, asymmetrical V-bends and step-bends) is studied through four methods: simulation of the teeth movements with a typodont, measure of the forces and the moments with a mechanical system composed of pulleys and dead load devices, photo-elastic stress analysis, finite element stress analysis. Typodont experimentations display the dental movements: the easiest one is rotation; it is more difficult to observe distalization and mesialisation that occur with a version of the tooth. An experimental apparatus, composed of pulleys and dead load devices, allows to measure forces and moments released by a transpalatal arch, depending on the level of the activations. The authors notice that the distalization force for an asymmetrical V-bend activation is low, about 0.2 Newton for 5 degrees of activation. Photo-elastic stress analysis affords a visualization of the shear stresses induced by a transpalatal arch within the supporting structures of a root, thanks to the observation of colored fringes in a model constituted with two molars included in a birefringent resin. Meanwhile, the manipulation of the photo-elastic resin is not easy and the interpretation of the results remains difficult. The finite element method is a numerical analysis and consists in the fragmentation of the studied structure in a certain number of elements, where forces and moments can be applied: the three dimensional program analyses the stress and strains in these elements. Thus, we can: 1.--visualize strains and stresses of a transpalatal arch, when it is inserted in the lingual molar sheaths; 2.--observe the dental movements due to the activations of the transpalatal arch (rotation, version, mesialisation, distalization); 3.--study the stress induced in the periodontal tissue. The results of these studies are complementary and in agreement. Particularly, the second method (measure of forces and moments) and the finite element method provide a mathematical and visual model that could explain clinical results: thus, the main purpose of this research is to understand the orthodontic mechanics used in the edgewise or segmented technique. PMID- 11039272 TI - [Stability of orthodontic treatment of occlusal asymmetry]. AB - A common finding in orthodontic patients is asymmetric occlusion. These asymmetries can be dental, skeletal, or functional in origin. Since many patients have typical posterior overjet the use of Class II-Class III and anterior crisscross elastics are contra-indicated. Even in skeletal discrepancies axial inclination compensation can produce relatively normal overjet in the arch. The best strategy for non-extraction therapy is to move teeth around the arch rather than an en-masse movement of the entire arch. A number of methods for unilateral distalization are discussed. Midline correction requires the determination of facial, apical base, and posterior midpoints. Differential mechanics between patients with apical base discrepancies and no apical base is presented. Although intermaxillary elastics can be indicated the undesirable effects of eruption and frontal occlusal plane tilt should be considered. Advantages in control and ease of occlusal correction rest with intra-arch mechanics. The use of intermaxillary elastics for the correction subdivision cases can lead to instability and or mandibular shifts. PMID- 11039273 TI - [Orthodontic treatment and joint stability. Disk luxation and mandibular path. Part 1]. AB - Jaw equilibrium is one of the most important goal to achieve, during orthodontic treatment. The clinical diagnosis of discal dislocations allows to track them systematically and avoids many dysfunctions which are responsible of relapses. PMID- 11039274 TI - [Orthodontic therapy and joint stability. Clinical diagnosis of disk luxation. Part 2]. AB - Discal dislocations are not enough defined by the mandibular opening diagram. Understanding the anatomical phenomena surely helps to improve diagnosis. However, localizing the medial sagittal plane and the incisor midlines is of real importance to develop solutions, from a treatment standpoint. PMID- 11039276 TI - [Overbite patterns: therapeutic implications]. AB - Orthodontic cases, presenting a deep-bite skeletal pattern, show a number of structural and alveolodental features. In order to reveal the areas of imbalance, a study based on the principles of Biggerstaaf's cephalometric analysis was performed: this analysis allows a more precise approach than the classical angular analysis, showing up the proportions of each patient and their variations. Consequently, it remains possible to assert a treatment strategy. PMID- 11039275 TI - [Complications of nasal obstruction in children]. AB - The author reviews the morphogenetic and functional consequences of oral breathing in childhood. PMID- 11039277 TI - [Growth: responses to pathology. Preliminary study]. AB - Many pathologies involve face. Among them, many have craniofacial growth consequences. The authors' aims are to analyze some of these pathologies, where clinical observations emphasize the role of the different craniofacial growth patterns. Despite a complex relation between the malformative part and the deformative one, untreated observations provide a better understanding of some craniofacial growth defects. Syndromes can be classified in 4 categories, involving the primitive causes of the clinical finding: 1. organic abnormalities of one or many functional matrix, 2. localized abnormalities of the anatomical structures, 3. general abnormalities of the conjunctive tissue, 4. mixed syndromes. Many observations will presented. PMID- 11039278 TI - [Class II mechanics and overbite]. AB - Class II mechanics are supposed to affect the vertical balance of the face when they are used in hyperdivergent patterns. We carried out a study about class II mechanics effects on 100 patients; 50 of them are hyperdivergent (GoGn/SN > or = 37), and 50 non-hyperdivergent. They all were treated with an Edgewise appliance without any high pull head gear. The authors showed that GoGn/SN angles and Occ/SN tend to be lessened in a skeletal open-bite population. Thus, Class II mechanics can be prescribed in any vertical facial pattern. PMID- 11039279 TI - [New anesthetics]. AB - Since the introduction of cocaine local analgesia in 1886, and the subsequent development of procaine (1904) and other closely related ester-type compounds, dentistry has prided itself on being as close to 'painless' as possible. In the late 1940s the newest group of the local anesthetic compounds, the amides, was introduced. The initial amide local analgesic, lignocaine (Xylocaine), revolutionised pain control in dentistry worldwide. In succeeding years other amide-type local anesthetics, mepivacaine, prilocaine, bupivacaine and etidocaine, were introduced. They gave the dental practitioner a local anesthetic armamentarium which provided pulpal analgesia for periods of from 20 minutes (mepivacaine) to as long as three hours (bupivacaine and etidocaine with adrenaline). In addition these popular drugs proved to be more rapid-acting than the older ester-type drug and, at least from the perspective of allergenicity, more safe. In 1976, in Germany, the newest amide local analgesic, carticaine HCl was introduced into dentistry. Articaine (the generic name was changed) possesses properties similar to lignocaine but has additional properties which made the drug quite attractive to the general dental practitioner. In 1986 articaine was introduced in North America (Canada) where it has become the most used local anesthetic, supplanting lignocaine. Articaine has been approved for use in the United Kingdom. In this introductory discussion we review the development of articaine and discuss its place in the dental local analgesic armamentarium. PMID- 11039280 TI - [The possible secondary effects in cases of local anesthesia]. AB - Local anesthetics are the safest and most effective drugs for pain control. Over 300 million local anesthetic cartridges are administered by dentists in the United States of America annually, yet serious complications reported number but a handful. Complications are categorised as localised or systemic. Localised complications arise at the site of needle penetration or anesthetic administration while systemic complications involve the entire organism. Localised complications include needle breakage, paresthesia, trismus, haematoma and facial nerve paralysis, while systemic complications are psychogenic to the act of receiving an injection, allergy and drug overdose (toxic reaction). These potential complications are briefly described in the following paper. PMID- 11039281 TI - [Pain perception, mechanisms of action of local anesthetics and possible causes of failure]. AB - First, the fundamentals of impulse transmission and pain perception are revised. The role of the primary afferent nociceptors is explained. Dental pain is described as a form of acute pain and the mechanism of nociception is fundamental. Peripheral and central sensitization can evolve. The second part covers the pharmacological aspects. Local anesthetics reduce impulse transmission by interfering with the mechanism of normal depolarisation. Binding to specific receptors located at the nerve membrane, more specifically on the sodium channel, results in decreased or eliminated permeability to sodium ions and leads to interruption of nerve conduction. The different types of local anesthetics used in dentistry are discussed in more detail with respect to their physico-chemical characteristics and analgetic properties. The importance of factors such as lipophilicity, degree of protein binding and dissociation constant pKa are explained together with the clinical implications of pH and possible toxic effects. Failure of local anesthesia can be the result of problems with the administration of the product or can have a pharmacological basis. Injection of the anesthetic should take place in amounts large enough, with suitable volume and as close as possible to the nerve. When infection and inflammation are present, the intravascular resorption of the anesthetic will accelerate and the lowered pH influences diffusion negatively. Repetitive administration can induce the phenomenon of tachyfylaxis (decreased anesthetic effect). PMID- 11039282 TI - [What is the cause of failure of local anesthesia?]. AB - Local anesthesia fails in 10% of cases of inferior alveolar nerve block and 7% of all cases of local anesthesia in general practice. Possible causes of failure are infection, wrong selection of local anesthetic solution, technical mistakes, anatomical variations with accessory innervation and anxiety of the patient. In this publication we discuss reasons for frequent failure in case of infection and in inferior alveolar nerve block. Understanding the mechanism of failure in local anesthesia, makes it possible to formulate guidelines to guarantee success. These measures are discussed in detail. PMID- 11039283 TI - [The problem of cross-infection during the administration of local anesthesia]. AB - Serious infections can readily be transmitted in the dental situation when percutaneous injuries occur. In most cases needlestick injuries are responsible. In this way, dentists are at risk of professionally acquired infectious diseases with high morbidity and mortality, such as Human Immunodeficiency disease, Hepatitis B or C. Today, immunisation is only possible for HB virus and not for the other bloodborne infectious micro-organisms discussed. The transmission risk is influenced by the type and number of micro-organisms present in the blood, presence of visible blood on the needle, depth of the injury and size and type of needle used. Prevention is of paramount importance. Increased emphasis in the prevention of percutaneous injuries as part of infection control training is indicated. PMID- 11039284 TI - [Local anesthesia in children]. AB - Local analgesia in children necessitates a correct approach. Sufficient explanation about the procedure and preparation of the child are important. Because of differences in anatomy, the technique of anesthesia needs adaptation. The different techniques are reviewed and discussed. The children are at greater risk of overdose when local anesthetics are administered. Calculation of the maximum permitted dose is necessary and with good precautions accidents can be prevented. PMID- 11039285 TI - [Local anesthesia and patients presenting with medical pathologies; the use of anamnesis in the prevention of medical complications in the dental office]. AB - The treatment of medically compromised patients is becoming a frequent event in general dental practice as a consequence of the longer presence of natural teeth, an increase in life expectation and a shift towards more ambulatory medical treatment of patients with chronic diseases. Since the dentist is responsible for a correct approach of these patients, it is important that he has knowledge of interactions between medical conditions, compensating mechanisms and medical treatment and the impact of dental treatment on this. In this context, medical history taking is useful. The medical history should be taken as correct as possible, with careful interpretation of the information. Necessary preventive measures should be known. PMID- 11039286 TI - [Should the dentin smear layer be preserved or eliminated? (Review of the literature)]. AB - The smear layer is a direct consequence of instrumentation of the root canal wall. Hand instrumentation as well as ultrasonic instrumentation produce a smear layer on the canal wall. This smear layer is composed of dentine, remnants of pulp tissue and odontoblastic processes and bacteria in an infected canal. Removal of the smear layer is accomplished by the irrigation of root canals with EDTA (17%) followed by NaOCl (5.25%). Permeability of the dentine is increased by the removal of smear layer. In this way the bacteria within the infected tubuli can be more easily destroyed by an intracanal dressing. Whether the removal of smear layer decreases the apical leakage is uncertain. To establish the clinical consequences from removal or preservation of the smear layer, further research is needed. PMID- 11039287 TI - [Iatrogenic risks during the obturation of the root canal system]. AB - A hermetic seal is the main goal of obturation of the root canal system. For that purpose guttapercha still remains the filling material of choice. Most filling techniques use plasticized guttapercha and a condensation technique to fulfil the requirements of hermetic seal. The heat to plasticize and the forces applied to condense that material may have an influence on the periodontal ligament and/or the remaining tooth structure. Furthermore can overfilling of guttapercha and more likely the sealer, affect the periodontal tissue. Although rare, mandibular nerve damage and adverse effects on the sinus maxillaris can be the result of inadequate treatment. PMID- 11039288 TI - [Consequences of dental restorative treatment (and of the dental materials used in its execution) on pulp tissue]. AB - Pulp tissue may suffer from carious invasion, direct or indirect trauma and last but not least from dental treatment. As the general population grows older and retains longer its dentition, teeth will inevitably experience a complex history of multiple episodes of aggressions. It is therefore not surprising that the dental practitioner is confronted more and more with teeth reacting acutely or chronically (in other words, painfully or not) after proper dental treatment. This paper describes the common pathologic and iatrogenic events that may have an effect on the pulpal health status. Preventive endodontic treatment is to be considered in cases of extensive dental treatment. PMID- 11039289 TI - [Intramedullary compression nailing of long tubular bones]. PMID- 11039290 TI - [Intramedullary compression nailing of long tubular bones]. AB - The biomechanical principle of intramedullary compression osteosynthesis is based on the implantation of a movable intramedullary nail that is statically interlocked in distal round holes and dynamically interlocked in a proximal slot. Distraction of the nail against the proximal interlocking screw by means of a compression screw leads to a relative movement of the proximal fragment directed distally against the nail. This results in direct contact of the main fragments under increasing compression. Simple fractures, pseudarthroses and osteotomies within the diaphyses of the long bones represent promising indications for the use of compression nailing. Furthermore, this method enables extraordinarily stable knee and ankle arthrodeses. Major positive aspects are controlled adaptation of fragments and a significantly increased stability of the fracture as compared to conventional intramedullary nailing techniques, especially as rotational forces are concerned. The biomechanical advantages result in earlier full weightbearing and an increased rate of fracture union in delayed healing. Given the use of optimized implants and instruments, compression intramedullary osteosynthesis offers a remarkable potential for further improvement in both the spectrum and success of intramedullary nailing. PMID- 11039291 TI - [Morphological changes in arterial ruptures]. AB - The historical opinion that the intima layer of a ruptured artery of muscular type could stop bleeding by "rolling in" should be controlled experimentally. Five segments of human femoral/popliteal artery (3-4 cm long) were overstretched until complete rupture occurred. Furthermore a longitudinally split and a partially oblique incised segment were ruptured. As morphological finding a sandclock deformity was observed in the region of rupture. This phenomenon was induced by adventitia layer, which closed the ends of ruptured media layer like a Chinese finger trap. In the longitudinally split segment a transverse rupture of the media layer could be observed, while the fibers of adventitia layer were pulled out when the traction was continued. Neither macroscopical nor microscopical signs could be found for "rolling in" of intima or media layer. The reason for spontaneous hemostasis after arterial rupture is more likely the activation of platelets by collagenous fibers of adventitia layer than "rolling in" of intima or media layer. If there is no finger trap mechanism of adventitia layer like in shot- or stab wounds a massive blood loss must be expected. PMID- 11039292 TI - [Timing of the surgery of rupture of the anterior cruciate ligament. Effects of acute or delayed surgery on arthrofibrosis rate and work disability]. AB - The optimal time to perform acute ACL reconstruction with respect to arthrofibrosis is discussed. Most authors prefer delayed surgery. The definition of the term "acute" varies between 48 hours and 4 weeks. In this study the limit was set at 60 hours. Acute ACL reconstruction was performed in 39 patients and delayed surgery in 35 patients after they had regained full ROM. The incidence of arthrofibrosis was not higher in the acutely operated group whereas overall inability to work was 44% lower in this population. When the indication is clear, we think that acute ACL reconstruction may be performed within 60 hours without a higher risk of postoperative development of arthrofibrosis. Nowadays, this strategy should also be considered for economic reasons. PMID- 11039293 TI - [The Balser plate with ligament suture. A dependable method of stabilizing the acromioclavicular joint]. AB - Between June 1990 and December 1997 116 patients with complete dislocation of the acromio-clavicular joint were treated operatively. According to the classification of Tossy and Rockwood all patients had type Tossy III or Rockwood III, IV and V of lesion. The retrospective part of the study from June 1990 to August 1994 represents 48 patients treated in 31 cases with wire-cerclage, in 14 cases with PDS-cerclage and in 3 cases with a combination of Kirschner wires and PDS. In a second group between September 1994 and 1997 68 patients were treated operatively with a special hook-plate, called Balser-plate, combined with suture of the corakoclavicular ligaments, the articular capsule and the intraarticular discuss. In the first group there was a postoperative immobilisation of the injured shoulder for 2,3 weeks necessary; in comparison to non immobilisation at the Balser-plate group. The range of motion in the Balser-plate group was free up to 90 degrees abduction. The removal of implants was performed in both groups after approximately 3 month. Postoperative complications were 8 reluxations at the acromio-clavicular joint and 10 superficial infects at the Non-Balser group and 4 superficial infects and 2 subcutaneous haematoma in the Balser group. We saw no reluxation in the Balser group. We examined 30 of 48 patients of the Non Balser group after average 50.1 month and 57 of 68 patients of the Balser group after 24.6 month. We compared the functional result, a questionnaire and the ultrasound examination of the acromio-clavicular joint with and without 10 kg weight bearing of the arm. 87.7% of the Balser patients and 67.7% of the Non Balser-patients had free movement of the injured shoulder. Another 14.2% and 11.4% of the Balser group complained on shoulder pain with weight bearing and extreme moval in comparison to 17.4% of the Non-Balser group. At ultrasound examination comparing the injured to the non injured arm with and without 10 kg weight bearing there was a clavicula-elevation of 0.3 mm and 0.6 mm at Balser patients and 3.0 mm and 2.5 mm at Non-Balser-patients for the injured side. In normal position the acromio-clavicular width was physiological in 50.2% of Balser patients in comparison to 36.6% of Non-Balser-patients. Under weight bearing the acromio-clavicular width increases in both groups. 63.2% of the Balser-group patients are satisfied with the result of operation, but only 43.3% of the other group (p < 0.05). Completely dissatisfied were 20% of the Non-Balser group, particularly because of the bad cosmetic result (40%). PMID- 11039294 TI - [Stable and unstable pertrochanteric femoral fractures. Differentiated indications for the dynamic hip screw]. AB - The aim of all surgical procedures in the treatment of trochanteric fractures in elderly and even geriatric patients is achievement of initial stability. We examined in a clinical trial whether primary stability was achieved in all types of trochanteric fractures following osteosynthesis with the Dynamic Hip Screw (DHS). From 1994 to 1996, 122 patients with trochanteric fractures had osteosynthesis by dynamic hip screw. Patient records were evaluated and all data got registered with a standardized protocol;clinical radiological outcome was analysed after an average period of 1.9 years after injury according to the Traumatic Hip Rating Score. 22% of all patients died meantimes, 51.6% of the remaining 95 patients could get examined. The average age was 75.5 years, the patient population showed an increased preoperative morbidity (2.5 points) according to ASA-Score. 81% showed progressive osteoporosis. According to the AO classification 47% stable fractures (type A-1) and 53% instable trochanteric fractures (type A-2 and A-3) occurred. Surgery lasted 77 minutes average in osteosynthesis of stable fractures. The duration of 108 minutes in instable fractures was significantly higher, as well as the blood loss was 43% increased in these complex fractures. Complications closely associated to the osteosynthesis appeared only in instable fractures (7%). Also common complications (24.6%) predominated with 15.6% in type A-2 and A-3 fractures versus 9% in type A-1 fractures; mortality was also different with 5.7% versus 1.6%. Assessment of the functional outcome according to THRS showed a significant deterioration of 20 points in 71% of all patients compared with the preoperative score. The results show that dynamic hip screw osteosynthesis in instable trochanteric fractures is associated to a higher incidence of complications. While the dynamic hip screw still represents the standard implant in stable fractures of the trochanteric region, being aware of improved intramedullary implants regarding biomechanical features and surgical technique, the results justify to critical consider the use of DHS for osteosynthesis in instable fractures of the trochanteric region. PMID- 11039295 TI - [Metaphyseal defect substitute: hydroxylapatite ceramic. Results of a 3 to 4 year follow up]. AB - 45 trauma patients with fracture-related metaphyseal bone defects filled with hydroxyapatite ceramic (Endobon) were followed for three to four years after surgery. Osseointegration of Endobon could be confirmed histologically in all biopsates, which could be obtained during scheduled reintervention for hardware removal. The objective of clinical and radiological follow-up was to evaluate the extent of ceramic remodeling as well as long-term implant outcome. Thirteen patients undergoing surgery in 1992 or 1993 were available for follow-up. None of these patients showed radiologic evidence of loss of bony reconstruction or HAC sintering, nor was there evidence of HAC resorption or fragmentation. Our results -with respect to the examined few patients--suggest that Endobon is useful for filling bony defects in carefully selected trauma patients if certain surgical technique prerequisites are met. Like cancellous autografts special alloplasts appear to go some way toward solving problems of defect filling encountered in clinical practice. PMID- 11039296 TI - [Ultrasound characterization of burn scars in children]. AB - So far, little is known about the acoustic phenomena of high-frequency sonography for the assessment of healing processes in thermal wounds. However, ex vivo investigations have shown clear age-dependent differences in the rate of sound propagation in thermal scars compared to healthy skin. In order to answer the question of whether measurable acoustic characteristics of burn or scald scars can be classified in a way which corresponds to the clinical severity of the injuries, age of scar or type of treatment, 92 thermal scars, with an average scar age of 3.1 years, in 55 children were investigated with regard to corium thickness and echogenicity. A control group of 25 non-injured children of similar age was studied for comparison. It was shown that measurable echogenicity differences can allow conclusions about clinical severity to be drawn. The comparison of different therapeutic strategies showed no therapeutic effect either for compression or for the application of silicone gel sheets. In contrast, a decrease in scar thickness and a loosening of scar structure could be seen after early application of a sterile, silicone-covered polyamide net bandage. The 20-MHz-Sonography is a suitable non-invasive procedure for the characterisation of burn scars. PMID- 11039297 TI - [Functional results after suture repair in ruptures of the long biceps tendon with special consideration of subacromial impingement]. AB - Operative treatment for ruptures of the long biceps tendon still is discussed controversially. In the present literature the keyhole-technique is recommended according to favourable biomechanical conditions. In recent years refixation to the short biceps tendon was preferred. Now it is supposed that this technique may provoke subacromial impingement considering the loss of depression function of the long biceps tendon to the humeral head. Between 1980 to 1991 83 patients with rupture of the long biceps tendon were treated operatively by refixation to the short head. 28 patients were investigated after an average follow-up of 6.5 years. Due to the criteria of the Constant-Score 85% of patients achieved very good, 15% good results. At our patients provocation of a subacromial impingement could not be observed. The subacromial space was not reduced in the postoperative x-ray control. Compared with the non-operated shoulder isokinetic determination of isometric maximal peak torque for elbow-flexion, shoulder-abduction and shoulder-flexion yield to almost identical results for the operated shoulder. Refixation to the short head can be advised for treatment of ruptures of the long biceps tendon due to the certain technique with a low complication rate and very good functional outcome. PMID- 11039298 TI - [Fracture of the sacrum]. PMID- 11039300 TI - [Transoral correction osteotomy in transdental fracture dislocation that healed in the wrong position]. AB - Undislocated odontoid fractures may lead on the basis of conventional x-rays only to a wrong conclusion with regard to biomechanical aspect of stability. In this aspect the classification based on Anderson and D'Alonzo takes a high risk to misunderstand the fracture stability and can results in a secondary fracture dislocation. Therefore it is important to make the decision about operative versus nonoperative treatment on the base of the trauma mechanism. In this case report we elucidate this problem and the higher risk of anterior approach for correction. Furthermore a better classification of dens fractures will be recommended. PMID- 11039299 TI - [Ventilation in prone position in a 5-year-old child after multiple trauma. Effective treatment of persistent atelectasis]. AB - We report on the ventilation in prone position in a 5-year-old traumatized child with severe thoracic and abdominal injuries (lung contusion, rib fractures, rupture of liver and spleen). Under continuous analgesic sedation, the young patient was ventilated in prone position for 6 h, since acute lung injury and atelectasis persisted despite various therapeutic measures (artificial ventilation in the pressure controlled mode, fiberoptic bronchoscopy, reexpansion maneuver). After initiation of the prone position, we observed a rapid increase in arterial oxygenation, which persisted in the following period. The hemodynamic situation remained stable. The complete disappearance of atelectasis was demonstrated radiologically after supine repositioning. After cessation of analgesic sedation, the extubation was performed 2 days later. Furthermore, we found no side effects of the prone position on the injured abdomen, and the liver function improved rapidly. Although there is a lack of experience with ventilation in prone position in pediatric intensive care, our report might be a recommendation for the indication of this technique in children. PMID- 11039301 TI - [Fatal soft tissue infections after arthroscopy of the knee joint. A diagnostic or therapeutic problem?]. AB - After detection of a bacterial infection of the joint, an absolute indication for intervention is given. Systemic antibiotic drug therapy is indicated and drainage of the joint has to be performed immediately. The following therapeutic algorithm regimen is a proven remedy in treating pyoarthrosis of the knee joint: During the initial period, the infection can be controlled by arthroscopic irrigation and systemic antibiotic therapy. Depending on the progredient findings or if the symptoms persist, curing the joint by open synovectomy is the next step of treatment. If open joint revision including synovectomy is not performed or is performed too late, there will be a threat of irreversible damage of the afflicted joint up to septic spread endangering the patient's life. We report on two patients suffering from generalized sepsis resulting in death after delayed therapy for knee joint infection. Regarding the presented cases, it can be concluded that indication to early surgical joint debridement including open synovectomy of the knee is still rarely seen after development of pyoarthritis. PMID- 11039302 TI - [Report on the current status of planned structure changes in surgery, accident surgery and orthopedics]. PMID- 11039303 TI - [Orthopedics--orthopedic surgery--accident surgery. Is the circle closed?]. PMID- 11039304 TI - Effects of nerve growth factor on antioxidative system in the thalamus of MPTP treated Wistar rats. AB - 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced parkinsonism is one of the most useful models for the study of that disease. It has been suggested that MPTP-induced neurotoxicity may involve the production of reactive oxygen species. MPTP was applied intracerebrally, unilaterally, in the striatum in single dose of 0.09 g/kg b.w. The second group was treated both with MPTP and nerve growth factor (NGF) in dose of 7 ng/ml. NGF was applied immediately after the neurotoxin. Control group was treated with 0.9% saline solution in the same manner. Animals were decapitated 7 days after the treatment. In the group treated with MPTP, the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) was decreased in ipsilateral thalamus, compared to control values as well as to the contralateral thalamus. In the same structures superoxide anion production was increased, compared to controls. Following the application of both MPTP and NGF, the activity of SOD and GSH-Px remained on control values, while the superoxide anion content was decreased, compared to controls. These results indicate a temporal and spatial propagation of oxidative stress and spread protective effects of NGF on the thalamus, the structure that is distant, but very tightly connected with striatum, the place of direct neurotoxic damage. PMID- 11039305 TI - Cytochrome C oxidase activity and total glutathione content in experimental model of intracerebral aluminum overload. AB - Treatment of Wistar rats with aluminum chloride causes astroglial and neuronal cell damage in the selective brain regions of association cortex and hippocampus, seen in patients with Alzheimer's disease. Adult Wistar rats were treated with unilateral intrahippocampal injection of AlCl3 in one single dose of 3.7 g/kg b.w. Control group of animals was treated with 0.9% saline solution likewise. Animals were sacrificed by decapitation seven days after the treatment. Activity of cytochrome C oxidase (COX) and total glutathione content were measured in the ipsi- and contralateral hippocampus and forebrain cortex. Activity of COX was mutually decreased in the hippocampus (ipsi- 30%, contra- 34%), as well as in the forebrain cortex (ipsi- 44%, contra- 47%), compared to controls. These decrease could indicate a deficiency in reducing equivalents with concomitant altered proton gradient and function of electron transport chain, as well as decreased ATP synthesis. Content of glutathione, a clue antioxidative factor, was decreased for about 50% in all examined structures, primary suggesting an impaired regeneration of reduced glutathione. Such distribution of diminished antioxidative defense could be the consequence of the specific brain distribution of transferrin receptors, which was also a main protein carrier for Al. Furthermore, at the cellular level Al could impede glycolysis with consequent decreased production of reducing equivalents which were necessary for glutathione synthesis/reduction, as well as for proton gradient and functionality of electron transport chain. PMID- 11039306 TI - [Indications and results of fasciotomy in vascular injuries of the lower extremities]. AB - Aim of this study was to re-evaluate the indications for fasciotomy in war vascular injuries of the lower extremities. Retrospective and partially prospective analysis of 31 patients with surgical revascularisation performed during 1999 was done. Fasciotomy has been used as a prophylactic measure against development of Compartment Syndrome (CSy) in three out of ten patients within the first group where ischemia time was less than six hours before the time of repair. The second group, where ischemia time was longer than six hours before the time of repair, prophylactic fasciotomy (measured compartmental pressure lower than 30 mmHg) was performed in 8 patients. In 13 patients with already developed CSy fasciotomy was performed as the delayed treatment (measured compartmental pressure higher than 30 mmHg). Neither one of patients from the first group developed CSy. All patients who developed CSy had necrosis of neuromuscular tissues at the time of surgery. Musculectomy was required in five and limb amputation in six patients. The conclusion of this study is that when the ischemia time is less than six hours before the time of repair fasciotomy is not necessary. When the period from injury to the revascularisation is longer than six hours the prophylactic fasciotomy is recommended. PMID- 11039307 TI - [Manifestations of epileptic seizures after acute cerebrovascular disease]. AB - The study included 80 patients hospitalized in Intensive Care Unit, in whom was with confirmed the occurrence of seizures after acute cerebrovascular diseases. They were classified according to sex, age, etiopathogenesis of stroke, latence from the onset of cerebrovascular disease to the occurrence of first seizure, hemispheric and intrahemispheric localization of cerebrovascular lesion, and clinical types of seizures. PMID- 11039308 TI - [Importance of determining the absolute CD3+ lymphocyte count during induction therapy with antithymocyte globulin in kidney transplantation patients]. AB - Antithymocyte globulin (ATG) is successfully applied in prophylaxis and treatment of renal allograft rejection. However, it is an expensive mode of therapy, associated with increased risk of opportunistic infections and lymphoproliferative diseases. For this reason, monitoring of ATG immunosuppressive effects as well as individual dose adjustment represent an important therapeutic approach. Here we report our results of ATG dose titration according to total lymphocyte count (< 300/microliter) and absolute CD3+ count (< 50/microliter) in seven renal transplant patients. Monitoring of absolute CD3+ count enabled reduction of the mean daily dose from the recommended dosage in all patients. Our results have also shown that the absolute CD3+ count is a more reliable parameter than the total lymphocyte count for monitoring of ATG biological effects on T cells. When rapid, significant and stable decrease of absolute CD3+ count is reached, ATG dose can be further adjusted according to the total lymphocyte count. With this approach, ATG treatment becomes rational and safe, with well established immunosuppressive effect, reduced risk of overimmunosuppression and considerable cost benefit. PMID- 11039309 TI - [Registration of the silent period in masseter muscles during study of the mandibular reflex]. AB - Under the application of clinical electromyography, nowadays is more widely used reflex investigation, i.e., the investigation of the reflexes to the extension. In stomatognathic system is the most frequently applied the investigation of mandibular reflex. Silent period was registered during the investigation of mandibular reflex. Silent period is the reflex pause or the period of suppressive activity that is subsequent to some sort of stimuli during or following muscle contraction. In current dentistry, silent period, registered by electromyography, represents an important parameter in functional diagnosis. The aim of the study was to establish the duration of silent period in masseter muscles during the investigation of mandibular reflex in edentulous patients with the pair of newly made full dentures and in the subjects with natural healthy intact dentition, respectively. The investigation included 20 subjects of average age 44 years. The results of registered silent period during the investigation of mandibular reflex in edentulous patients had the lower values (18.21 ms) than in the subjects with healthy intact dentition and with a pair of newly made full dentures (20.51 ms), respectively, which have confirmed the physiologic time of the duration of silent period. PMID- 11039310 TI - [Effect of aging on visual evoked potentials]. AB - Experimental and clinical studies, aiming to establish the influence of aging on latence, amplitude and types of waves of visual evoked potentials (VEP) are numerous. It was concluded that P-100 latence prolonged with aging, particularly after the age of 55, while P-100 amplitude had been gradually decreased during the life. During the aging, senile changes in the eye and optic nerve (for example, senile miosis, degenerative retinal changes, geniculostraital defect) can be reflected to the changes in VEP (P-100). Aim of our study was to establish if the aging influences the alteration in latence and wave amplitude in VEP, as well as to standardize normal values of VEP in different ages. VEP was registered in 135 subjects classified into 3 groups. Group I comprised 49 subjects, aged 17 30 (average age 20.92 years), group II comprised 34 subjects aged 31-54 years (average 42.55 years). Group III--over 55 years (average age 69 years) comprised 52 subjects. There were 55 females and 80 males. Results have shown that the mean value of amplitude for both eyes in the first two groups of younger subjects was 9.03 microV and 8.91 microV, respectively, while in the group of older subjects, it was slightly decreased--8.54 microV. P-100 latence prolonged insignificantly until the age of 55, and afterwards in every 5 years it increased for 1.5 ms. Amplitude was insignificantly lower latence in VEP was insignificantly prolonged until the age of 55, and afterwards, the increase was significant. Amplitude was decreased insignificantly, gradually during the life. PMID- 11039311 TI - [Cyclosporin A in the therapy of nephrotic syndrome caused by focal-segmental glomerulosclerosis]. AB - The use of Cyclosporin A (CsA) in treatment nephrotic syndrome in the primary focal and segmental glomerulosclerosis (FSGS) is controversial. A prospective study was comprised of 10 adult patients with nephrotic syndrome caused by FSGS, treated with CsA and corticosteroids. Six patients were females, 4 males, aged 35 +/- 8 years. The daily urinary protein excretion ranged from 7 to 15 g/24 h (range 11.16 +/- 2.61 g/24 h). The follow-up interval lasted from 6-18 months. The serum CsA levels ranged from 80-120 ng/ml. Prednisone was administered orally 10-15 mg/day. Two months after the therapy onset, all patients experienced clinical improvement and reduction in the urinary daily protein excretion (range 9.74 +/- 2.14 g/24 h) with tendency to decrease, while a stable overall renal function was maintained. During 6 months, 6 patients were in remission (0.5 +/- 0.2 g/24 h), in two patients the proteinuria was retained 1.6-1.8 g/24 h and two patients had proteinuria 3.2-3.6 g/24 h. During the follow up period of this patients' group in the period of 18 months, the values of proteinuria were never above 3 g/24 h (1.15 +/- 0.9 g/24 h). These are encouraging results from initial treatment of FSGS with CsA and small doses prednisone. PMID- 11039312 TI - [Keratinocyte culture: clinical use and a model for research]. PMID- 11039313 TI - [Learning, remembering, memory--molecular biology aspects]. PMID- 11039315 TI - [Necrotizing fasciitis]. PMID- 11039314 TI - [Antisperm antibodies and their significance in the pathogenesis of female infertility]. PMID- 11039316 TI - [Lipedema complicated with lymphedema and chyloderma]. AB - Lipedema never reveals clinical picture of extreme lymphedema-elephantiasis, and skin signs and complications have not been observed. Aim of this paper is to present a case of lipedema with the initial lymphedema in which, after one episode of lymphangiitis and cellulitis, came to the rapid development of lymphedema followed by chyloderma. During the local treatment of extreme chyloderma with excessive exudation, semiocclusive synthetic dressings have been used for moist wound healing. The treatment was completed after 20 weeks with total epithelizsation, without maceration and irritation, without additional spreading of the chyloderma field, without wound infections, with fast and full relief of the pain. Lipedem with extreme lymphedema can be followed by skin complications of lymphedema like chylodermia. PMID- 11039317 TI - [The "woman" in military recruitment]. AB - Transsexuality is a phenomenon, which presents a rarity on recruitment. A case of male-to-female genuine transsexualism was presented in term of psychological psychiatric evaluation of a recruit for military service. A phenomenon of transsexualism is discussed through etiology, clinical picture, diagnosis, differential diagnosis and modern treatment. PMID- 11039318 TI - The folic acid analogue methotrexate protects frog embryo cell membranes against damage by the potato glycoalkaloid alpha-chaconine. AB - As part of an effort to improve the safety of plant foods, a need exists to more clearly delineate the mechanisms of toxicities of glycoalkaloids, which may be present in Solanum plant species such as potatoes, tomatoes and eggplants. Alpha chaconine is a major glycoalkaloid present in potatoes. To assess the possible influence of structure of pteridine derivatives on toxicity of potato glycoalkaloids, a previous study that demonstrated the protective effects of folic acid against the Solanum glycoalkaloid alpha-chaconine-induced toxicity on Xenopus laevis frog embryo cell membranes was extended to two folate analogues--a synthetic compound widely used as a therapeutic agent methotrexate, and naturally occurring L-monapterin. Adverse effects on embryos were evaluated by observing changes in membrane potentials with an electrochromic dye, di-4-ANEPPS, as a fluorescent probe for the integrity of the membranes. Methotrexate decreased alpha-chaconine-induced polarization, as did folic acid. This decrease may result from an alteration of membrane conformations that prevents the binding of the glycoalkaloid to the membrane receptor sites, and/or from effects on folic acid metabolism. In contrast, L-monapterin did not significantly reduce the alpha chaconine-induced toxicity. The possible significance of these results to food safety is discussed. PMID- 11039319 TI - Polymethoxyflavonoids from Vitex rotundifolia inhibit proliferation by inducing apoptosis in human myeloid leukemia cells. AB - Three polymethoxyflavonoids from the fruit of Vitex rotundifolia, namely 2',3',5 trihydroxy-3,6,7-trimethoxyflavone (Vx-1), vitexicarpin (Vx-5) and artemetin (Vx 6), were tested for their antiproliferative activity in human myeloid leukemia HL 60 cells. They showed a dose-dependent decrease in the growth of HL-60 cells. The concentrations required for 50% inhibition of the growth (IC50) after 96 h were 4.03 microM, 0.12 microM and 30.98 microM for Vx-1, Vx-5 and Vx-6, respectively. Treatment of HL-60 cells with the flavonoids induced morphological changes that are characteristic of apoptosis. We judged the induction of apoptosis by the detection of DNA fragmentation in agarose gel electrophoresis and the degree of apoptosis was quantified by a double-antibody sandwich ELISA and by flow cytometric analysis. The C-3 hydroxyl and C-8 methoxyl groups were found not to be essential for the activity, but the C-3' methoxyl instead of hydroxyl group lowered the antiproliferative and apoptosis inducing activity. These results suggest that the polymethoxyflavonoids isolated from V. rotundifolia may be used as potential chemopreventive and chemotherapeutic agents. PMID- 11039320 TI - A modified HET-CAM assay approach to the assessment of anti-irritant properties of plant extracts. AB - Hen's egg--chorioallantoic membranes were used to screen for and assess anti irritant properties among aqueous extracts of plants (HET-CAM tests), in connection with searches for plant-derived substances with topical anti-irritant action. The main question to be answered was whether CAM-assay screening of plant extracts could provide a useful route to identifying promising anti-irritant extracts for follow-up clinical testing. To be useful, the method would have to flag materials with strong anti-irritant properties, and would have to avoid registering false negatives. The tests conducted provided positive indications. We measured the delays in onset of three manifestations of membrane irritation vascular hemorrhaging, membrane lysis and membrane coagulation-observed with test substances relative to positive controls. Aqueous 15% lactic acid, a commonly used irritant in direct tests on human skin, was employed as the test irritant in this study. The ratio [irritation onset times after test substance pre treatment]:[onset times without test substance pretreatment] was used to measure the anti-irritant power of test substances. A scoring notation was devised for this which treats the delay parameters as independent effects. Most tested plant extracts showed no significant irritant or anti-irritant effects. Among the apparently anti-irritant plant extracts (approx. 10% of all those tested), most showed their greatest effect against hemorrhaging. Lesser but still readily measurable effects against membrane lysis and coagulation were also observed in nearly all the apparently anti-irritant extracts. Two of the tested extracts proved to be membrane irritants. Some key CAM assay results were compared with results obtained in direct tests on human skin using the same test irritant (15% lactic acid). In these comparative tests on skin, an essentially similar pattern of efficacy was obtained, with the plant extract deemed best in the CAM screenings, outperforming the benchmark anti-irritant hydrocortisone. From these initial results it appears that physiological CAM assays may prove useful in screening natural materials for anti-irritant properties, as alternatives to mechanism-dependent biochemical assays, or expensive direct screening tests on human subjects. Further work remains to extend the CAM screening approach to irritants other than lactic acid, and to assess its quantitative powers of prediction of topical anti-irritancy. PMID- 11039321 TI - A comparison of clinical, histopathological and cell-cycle markers in rats receiving the fungal toxins fumonisin B1 or fumonisin B2 by intraperitoneal injection. AB - Fumonisins B1 and B2 (FB1 and FB2) are fungal secondary metabolites produced by members of the genus Fusarium. Although FB1 is usually detected in greater quantities, FB2 frequently co-occurs in contaminated feeds and foods and contributes to the total toxin load. In the present study, the comparative toxicity of FB1 and FB2 was examined in male Sprague-Dawley rats administered toxin (0.75 mg/kg body weight) or vehicle control intraperitoneally (ip) for 2, 4 or 6 consecutive days. Clinical changes, including elevated serum cholesterol, alanine aminotransferase (ALT), creatinine and protein, were slightly more pronounced in FB1-treated rats. The most consistent hematological change was an increase in vacuolated bone marrow cells, which was more pronounced in FB1 treated rats. Histopathological changes were similar in FB1- and FB2-treated rats and included single cell necrosis in kidneys and liver, cytoplasmic vacuolation in adrenal cortex and lymphocytolysis in thymus. In the liver mRNA expression for the cyclin kinase inhibitor p21 gene was significantly increased in FB1- and FB2 treated rats, compared to controls. Expression of mRNA for the cyclin D1 gene was significantly depressed in FB2-treated rats. Hepatic cyclin E mRNA was elevated in response to FB1 and FB2 compared to controls. In FB2-treated animals this corresponded with decreased liver p27 mRNA expression. Hepatic proliferating cell nuclear antigen (PCNA) transcription was elevated in FB1- but not FB2- treated rats. Changes in liver microsomal protein levels of p27, cyclin E and PCNA were similar to changes in gene expression. In contrast, cyclin D1 protein levels were elevated in rats treated with FB1 and, to a lesser extent, FB2. The data indicate that FB1 and FB2 can alter the expression of genes associated with the cell cycle, and indicate a need for a further understanding of the mechanistic basis of FB1 and FB2 toxicity. PMID- 11039322 TI - Testing the potential of flaxseed to affect spermatogenesis: morphometry. AB - Quantitative information was collected on male reproductive effects of maternal and postnatal dietary exposure to flaxseed (20 or 40%), flaxmeal (13 or 26%) or standard NIH AIN-93 feed (0% flaxseed control). Measurements were made on the testes of F1 generation males rats (1) whose mothers were exposed to the diets designated above, and (2) who, after weaning, were placed on the same diet as their mothers for an additional 70 days. The seminiferous tubules comprised 86%, 84%, 84%, 84% and 85% of the total testis volume while the interstitial space comprised 12%, 14%, 14%, 14%, 13% of the total testis volume for the 0% flaxseed/flaxmeal, 20% flaxseed, 13% flaxmeal, 40% flaxseed and 26% flaxmeal groups, respectively. Statistically significant decreases in the absolute volume of the seminiferous tubules were observed in the 20% and 40% flaxseed-treated groups when these groups were compared to controls. Borderline statistically significant differences were also observed when Sertoli cell nucleolar number per tubular cross-section were compared in the 13% flaxmeal and 20% flaxseed treatment groups. These effects were not considered biologically significant because other parameters of male reproductive function appeared normal. Overall, the quantitative information obtained suggests that exposure to flaxseed/flaxmeal at the doses used in the present study does not adversely affect testis structure or spermatogenesis in the rat. PMID- 11039323 TI - Antimutagenicity of ethanol extracts of bee glue against environmental mutagens. AB - The antimutagenicity of ethanol extracts of bee glue (propolis) (EEBG) was evaluated, using Salmonella typhimurium strain TA98 as a test model, against two direct mutagens, 4-nitro-O-phenylenediamine (4-NO) and 1-nitropyrene (1-NP), and two indirect mutagens, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and benzo[a]pyrene (B[a]P) with S9 mix. EEBG was shown to suppress the mutagenicity of these compounds in a dose-dependent fashion. To delineate the mechanism of action of the antimutagenic effects of EEBG on the two indirect mutagens IQ and B[a]P, two possible points of blocking were considered: (1) cytochrome P-450 activity (route 1) and (2) interaction with microsome-generated proximate mutagens to generate an inactive complex (route 2). Our results clearly demonstrated, at a very low dose, remarkable suppression of the mutagenicity of both compounds by inhibiting either route 1 or route 2 pathway. Further studies indicated that EEBG was capable of inhibiting both the activities of hepatic cytochrome P-450 IA1-linked 7-ethoxyresorufin-O-deethylase (EROD) and IA 2-linked 7-ethoxycoumarin-O-deethylase (ECD) in a similar dose-dependent manner. Taken together, we demonstrated that EEBG was a good inhibitor for mutagenicity of direct mutagens, 1-NP and 4-NO, as well as for the indirect mutagens IQ and B[a]P in the presence of S9 mix via inactivation of microsomal enzyme activities (e.g. EROD and ECD) or antagonizing metabolic generation of the proximate mutagens of IQ and B[a]P. PMID- 11039324 TI - Assessment of the Pb and Cu in vitro availability in wines by means of speciation procedures. AB - The speciation of Pb and Cu in white and red table wines was investigated, in order to estimate their respective bioavailability to man. For this purpose, wines were subjected to in vitro gastrointestinal digestion, and the following properties were studied in the wines and in their gastric and intestinal digests: (1) the average conditional stability constant (Kav) of the strongest complexes (those inert to cathodic voltammetry) and of the respective ligand concentration (CCinert); (2) the distribution of the metal among the different bands of reverse phase high performance liquid chromatography (RP-HPLC) (groups of compounds of different molecular weight and/or polarity); (3) the total metal concentration and metal present in the soluble and in the dialyzable fractions of the digest. The CCinert of the red wines and the respective digests were much greater than those of the white wines and their digests. The conditional stability constants of the strongest soluble complexes after the digestion ranged between 5.9 and 6.1 for Pb. These parameters could not be determined for Cu. After the digestion the dialyzable metal fraction (a relative index of the metal potentially available for interaction with the inner biologic ligands) was only 16% of the total Pb in red wine, 62% in white Verde and 75% in white wine. For Cu the dialyzable metal fraction was 45% of the total metal in red wine, 64% in white Verde and 98% in white wine. PMID- 11039325 TI - Pubertal and postpubertal cadmium exposure differentially affects the hypothalamic-pituitary-testicular axis function in the rat. AB - The effects of administration of cadmium on levels of hormones along the hypothalamic-pituitary-testicular axis were studied in rats. Male rats were treated subcutaneously from days 30 to 60 (pubertal rats) or from days 60 to 90 of life (postpubertal rats), with cadmium chloride (CdCl2) at a dose of 0.5 or 1 mg/kg, every 4 days in an alternate schedule, starting from the lower dose. Age matched control rats received 0.3 m of saline subcutaneously every 4 days. The levels of norepinephrine (NE) increased on cadmium exposure in pubertal rats in all hypothalamic areas studied, but decreased in the median eminence. In contrast, in postpubertal rats the levels of NE only did not decrease in the posterior hypothalamus. Serotonin (5-HT) concentration in pubertal and postpubertal rats decreased in all hypothalamic regions, while serotonin turnover (measured by the ratio 5-hydroxyindolacetic acid/serotonin [5-HIAA/5-HT]) increased in the anterior hypothalamus. The serotonin metabolism was also increased in the median eminence in the pubertal and in the posterior hypothalamus in the postpubertal rats. Plasma levels of luteinizing hormone (LH) were not modified by cadmium in both age groups, but follicle stimulating hormone (FSH) levels decreased in postpubertal rats, but was not altered in pubertal rats. Plasma levels of testosterone increased in pubertal rats but decreased in postpubertal rats. Cadmium accumulation increased in the hypothalamus and testes in all the cadmium-treated animals, whereas in the pituitary accumulation of cadmium was found only in postpubertal rats. These data suggest that cadmium exerts age-dependent effects on the hypothalamic-pituitary-testicular axis function, and a disruption of the regulatory mechanisms of the hypothalamic pituitary-gonadal axis emerges. PMID- 11039326 TI - An assessment of chloramphenicol and thiamphenicol in the induction of aplastic anaemia in the BALB/c mouse. AB - The potential of the antibiotics chloramphenicol succinate (CAPS) and thiamphenicol (TAP) to induce aplastic anaemia in the female BALB/c mouse was investigated. CAPS was administered at 2000 mg/kg, and TAP at 850 mg/kg, daily by gavage, for 17 days. At 1, 13, 22, 41, 98 and 179 days after the final dose of each antibiotic, mice (n = 4 or 5) were sampled for haematological examination and haematopoietic stem cell assays. Both CAPS and TAP induced significant reductions in red blood cell count, haematocrit and haemoglobin values at day 1 post dosing; counts of colony-forming units-erythroid and colony-forming units granulocyte-macrophage, were similarly significantly decreased at this time. All these reduced parameters returned towards normal at days 13 and 22. At days 41, 98 and 179, results for all haematological values and stem cell assays in both CAPS- and TAP-treated mice compared with the controls; there was no evidence of a reduction in peripheral blood values or bone marrow parameters at the later sampling points, as would be expected in a developing or overt bone marrow aplasia. We therefore consider that the administration of CAPS and TAP, which have been associated with the development of aplastic anaemia in man, induce a reversible anaemia, but not a chronic bone marrow aplasia, when given at haemotoxic dose levels for 17 days in the BALB/c mouse. PMID- 11039327 TI - Attenuation of benzoyl peroxide-mediated cutaneous oxidative stress and hyperproliferative response by the prophylactic treatment of mice with spearmint (Mentha spicata). AB - The modulating effect of spearmint (Mentha spicata) on benzoyl peroxide-induced responses of tumor promotion in murine skin was investigated. Benzoyl peroxide (BPO) is an effective cutaneous tumor promoter acting through the generation of oxidative stress, induction of ornithine decarboxylase activity and by enhancing DNA synthesis. BPO treatment (20 mg/animal) increased cutaneous microsomal lipid peroxidation and hydrogen peroxide generation. The activity of cutaneous antioxidant enzymes, namely catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase, was decreased and the level of cutaneous glutathione was depleted. BPO treatment also induced the ornithine decarboxylase activity and enhanced the [3H]thymidine uptake in DNA synthesis in murine skin. Prophylactic treatment of mice with spearmint extract (10, 15 and 20 mg/kg) 1 hr before BPO treatment resulted in the diminution of BPO-mediated damage. The susceptibility of cutaneous microsomal membrane to lipid peroxidation and hydrogen peroxide generation was significantly reduced (P < 0.05 ). In addition, depleted levels of glutathione, inhibited activity of glutathione dependent and antioxidant enzymes were recovered to a significant level (P < 0.01, P < 0.05 and P < 0.01, respectively). Similarly, the elevated ornithine decarboxylase activity and enhanced thymidine uptake in DNA synthesis was inhibited significantly (P < 0.05 ) in a dose-dependent manner. The protective effect of spearmint was dose dependent in all parameters. The result suggests that spearmint is an effective chemopreventive agent that may suppress BPO-induced cutaneous oxidative stress, toxicity and hyperproliferative effects in the skin of mice. PMID- 11039328 TI - A comparison of the mainstream smoke chemistry and mutagenicity of a representative sample of the US cigarette market with two Kentucky reference cigarettes (K1R4F and K1R5F). AB - The incorporation of technologies into cigarettes such as filters, filter ventilation, porous cigarette papers, expanded tobacco and reconstituted tobacco sheet has resulted in cigarettes with a wide range of "tar" yields. The objectives of this study were to characterize the US cigarette market according to "tar" category (i.e. full flavor, FF; full flavor low tar, FFLT; or ultra low tar, ULT) and to determine whether the Kentucky reference cigarettes K1R4F and K1R5F are representative of FFLT and ULT cigarettes, respectively. As a means of characterization and comparison, the mainstream smoke from a representative sample of commercially available cigarettes from each market segment and the K1R4F and K1R5F Kentucky reference cigarettes was analyzed for the presence and level of 18 selected chemical constituents. In addition, a measure of the mutagenic activity of the mainstream smoke condensate from these cigarettes was determined using an Ames Salmonella mutagenicity assay. All cigarettes were smoked according to US Federal Trade Commission (FTC) guidelines. Results indicated that, overall, mainstream smoke constituent levels are well predicted by FTC "tar" yield--constituent levels increased as "tar" delivery increased. Based on the selected analytes measured in mainstream smoke, the K1R4F reference cigarette was generally representative of the FFLT segment of the US cigarette market. The K1R5F reference cigarette was representative of the ULT segment of the US cigarette market for cigarettes with "tar" deliveries approximate to it. In terms of mutagenic activity, a direct relationship was also demonstrated on a per cigarette basis-revertants per cigarette increased with increasing "tar" delivery. There was a weak tendency (R-square = 0.12, P = 0.08) for specific activity (revertants/mg "tar") to increase with decreasing "tar" yield-lower "tar" products had a slightly higher specific activity. No significant differences (P > 0.05) were observed when the specific activities of the condensates from the K1R4F and K1R5F reference cigarettes were compared to the market segments that they were designed to represent, FFLT and ULT, respectively. Overall, these results support the use of the K1R4F and the K1R5F as acceptable reference cigarettes for comparative mutagenicity and smoke chemistry studies of cigarettes available on the US market. PMID- 11039329 TI - Gellan gum. AB - For decades microbial exopolysaccharides have been invaluable ingredients in the food industry, as well as having many attractive pharmaceutical and chemical applications. Gellan gum is a comparatively new gum elaborated by the Gram negative bacterium Sphingomonas paucimobilis. Although its physico-chemical properties have been well characterized, the ecology and physiology of Sphingomonas, and the factors influencing the fermentation process for production of this gum have received much less attention. This review focuses on the metabolism and the enzymic activity of this bacterium, as well as the factors that influence gellan production, including process temperature, pH, stirring rate, oxygen transfer, and composition of the production medium. Potential strategies for improving the production process are discussed in the context of processes for the production of other microbial biopolymers, particularly exopolysaccharides. In addition, the importance and potential utility of gellan lyases in modification of gellan and in other applications is critically evaluated. PMID- 11039330 TI - Biosensors for the detection of organophosphorous pesticides. PMID- 11039331 TI - Imaging in stroke: the more you look, the more you see. PMID- 11039332 TI - MR imaging of brain tumors: toward physiologic imaging. PMID- 11039333 TI - MR angiography for the diagnosis of vasospasm after subarachnoid hemorrhage. Is it accurate? Is it safe? PMID- 11039334 TI - Vascular compliance in normal pressure hydrocephalus. AB - BACKGROUND AND PURPOSE: Normal pressure hydrocephalus (NPH) is considered to be a combination of altered CSF resorption and a reversible form of cerebral ischemia. The hypothesis tested in this study was that a reduction in venous compliance in the territory drained by the superior sagittal sinus (SSS) is associated with NPH and cerebral ischemia. METHODS: This prospective study involved 27 patients without evidence of hydrocephalus. This group was subdivided into those with normal MR findings and those with evidence of ischemia or atrophy. Ten patients with NPH then underwent MR flow quantification studies of the cerebral vessels. Five of these patients had the same studies performed after CSF drainage. Vascular compliance was measured in the SSS and straight sinus territory by use of MR flow quantification with net systolic pulse volume (NSPV) and arteriovenous delay (AVD) as markers. RESULTS: Vascular compliance of patients with ischemia or atrophy was significantly higher than that of healthy subjects (mean NSPV in the SSS, 417 microL and 274 microL, respectively). Patients with NPH showed lower compliance than that of the healthy subjects in the SSS (mean NSPV, 212 microL and 274 microL, respectively; mean AVD, 42 ms and 89 ms, respectively). After intervention, the NPH group showed compliance approximating the group with ischemia/atrophy. CONCLUSION: Vascular compliance is significantly different in the brains of healthy subjects as compared with that in patients with ischemia/atrophy or NPH. PMID- 11039335 TI - Normal pressure hydrocephalus: new concepts on etiology and diagnosis. PMID- 11039336 TI - Diffusion-tensor MR imaging of the human brain with gradient- and spin-echo readout: technical note. AB - Diffusion-tensor MR imaging of the brain is an objective method that can measure diffusion of water in tissue noninvasively. Five adult volunteers participated in this study that was performed to evaluate the potential of gradient- and spin echo readout for diffusion-tensor imaging by comparing it with single-shot spin echo echo-planar imaging. Gradient- and spin-echo readout provides comparable measures of water diffusion to single-shot spin-echo echo-planar readout with significantly less geometrical distortion at the expense of a longer imaging time. PMID- 11039337 TI - Diffusion-weighted imaging of patients with subacute cerebral ischemia: comparison with conventional and contrast-enhanced MR imaging. AB - BACKGROUND AND PURPOSE: The importance of diffusion-weighted imaging (DWI) for delineating acute ischemic lesions has been investigated extensively; however, few studies have investigated the role of DWI in the subacute stage of stroke. Because these lesions tend to appear bright throughout the first days of ischemia, owing to restricted diffusion, we speculated that DWI could also improve the detection of subacute infarcts as compared with conventional and contrast-enhanced MR imaging. METHODS: Interleaved echo-planar DWI with phase navigation was performed on a 1.5-T MR unit in a consecutive series of 53 patients (mean age, 66 +/- 14 years) with suspected recent cerebral ischemia. The interval between onset of clinical symptoms and MR imaging ranged from 1 to 14 days (mean, 6 +/- 4 days). Contrast material was given to 28 patients in a dose of 0.1 mmol/kg. RESULTS: DWI clearly delineated recent ischemic damage in 39 patients (74%) as compared with 33 (62%) in whom lesions were identified or suspected on conventional T2-weighted images. DWI provided information not accessible with T2-weighted imaging in 17 patients when evidence of lesion multiplicity or detection of clinically unrelated recent lesions was included for comparison. Subacute ischemic lesions were also seen more frequently on DWI sequences than on contrast-enhanced images (20 versus 13 patients). DWI was more likely to make a diagnostic contribution in the first week of stroke and in patients with small lesions or preexisting ischemic cerebral damage than was conventional MR imaging. CONCLUSION: Recent ischemic damage is better shown on DWI sequences than on conventional and contrast-enhanced MR images throughout the first days after stroke and may provide further information about the origin of clinical symptoms. Adding DWI to imaging protocols for patients with subacute cerebral ischemia is recommended. PMID- 11039338 TI - Effects of three different doses of a bolus injection of gadodiamide: assessment of regional cerebral blood volume maps in a blinded reader study. AB - BACKGROUND AND PURPOSE: Reconstruction of first-pass bolus information to derive regional cerebral blood volume (rCBV) maps is commonly performed in many centers; however, various protocols with different doses of paramagnetic contrast injections have been reported. We evaluated the dose dependency of rCBV maps in a brain tumor population by using three different doses of gadodiamide injection to evaluate their diagnostic accuracy in blinded reader sessions. METHODS: Eighty three patients with intraaxial brain tumors (72 gliomas) were studied at three centers and randomized to receive a bolus injection of 0.1, 0.2, or 0.3 mmol/kg per body weight of gadodiamide. rCBV maps were generated from T2*-weighted gradient-echo echoplanar sequences at 1.5 T. Data processing was performed according to the indicator dilution theory. RESULTS: The mean contrast-to-noise ratio (CNR) was significantly different between gadodiamide doses of 0.1 and 0.2 mmol/kg (CNR = 8.7 and 15.7) and between 0.1 and 0.3 mmol/kg (CNR = 17.7). No significant difference was found between doses of 0.2 and 0.3 mmol/kg. Sensitivity for the differentiation of benign and malignant brain tumors was 80%, 95%, and 91%, and specificity was 45%, 54%, and 43% by blinded readings at 0.1, 0.2, and 0.3 mmol/ kg, respectively, as compared with histologic findings. Nonblinded readings had a sensitivity of 83%, 100%, and 90% and a specificity of 82%, 100%, and 73% at 0.1, 0.2, and 0.3 mmol/kg, respectively. CONCLUSION: A dose of 0.2 mmol/kg of gadodiamide is recommended for reconstruction of rCBV maps if data are acquired with the T2*-weighted protocol described. PMID- 11039339 TI - Vasospasm after subarachnoid hemorrhage: diagnosis with MR angiography. AB - BACKGROUND AND PURPOSE: The possibility of treating intracranial vasospasm has increased the significance of its diagnosis and follow-up; however, so far, no ideal method is available. The goal of this study was to assess the accuracy of MR angiography versus intraarterial angiography (IA-DSA) in detecting vasospasm. METHODS: The study included 42 patients with acute spontaneous subarachnoid hemorrhage (SAH). Serial MR angiograms (minimum, two per patient within 10 days after the event; total, 149) were obtained prospectively using a 3D time-of flight technique covering the circle of Willis at 0.5 T. Forty-seven MR angiograms could be compared with intraarterial angiograms obtained within 24 hours of MR angiography. Vascular narrowing on both studies was rated consensually by two pairs of neuroradiologists using a scale from 0 (no narrowing) to 3 (severe narrowing). Categories 0 and 1 were considered an absence of vasospasm and categories 2 and 3 a presence of vasospasm. RESULTS: Agreement between MR angiography and IA-DSA (assessed with weighted kappa statistics) was substantial for the middle and anterior cerebral arteries (MCA and ACA) but moderate for the internal carotid artery (ICA). The sensitivity, specificity, accuracy, and positive and negative predictive values of MR angiography for detecting patients with vasospasm were 92%, 98%, 96%, 92%, and 98%, respectively. Considering each vessel separately, specificity was high for all locations (95 99%) and sensitivity was excellent for the ACA (100%) but poorer for the ICA (25%) and MCA (56%). CONCLUSION: MR angiography at 0.5 T is capable of identifying vasospasm after acute SAH but is less sensitive than IA-DSA for depicting vasospasm in the ICA and MCA. PMID- 11039340 TI - The role of MR angiography in the pretreatment assessment of intracranial aneurysms: a comparative study. AB - BACKGROUND AND PURPOSE: With developments in coil technology, intracranial aneurysms are being treated increasingly by the endovascular route. Endovascular treatment of aneurysms requires an accurate depiction of the aneurysm neck and its relation to parent and branch vessels preoperatively. Our goal was to estimate the clinical efficacy of MR angiography (MRA) in the pretreatment assessment of ruptured and unruptured intracranial aneurysms. We compared MRA source data (axial acquired partitions), multiplanar reconstruction (MPR) of these data, as well as maximum intensity projection (MIP) and 3D-isosurface images with intraarterial digital subtraction angiography (IA-DSA). METHODS: The study was performed in 29 patients with 42 intracerebral aneurysms. The MRA data were examined in four different forms--as axial source data, MPR images of the source data, and MIP and 3D isosurface--rendered images. A composite standard of reference for each aneurysm was then constructed using this information together with the IA-DSA findings by looking at aneurysm detection rate, aneurysm morphology, neck interpretation, and branch vessel relationship to the aneurysm. All techniques, including conventional IA-DSA, were then scored independently on a five-point scale from 1 (non diagnostic) to 5 (excellent correlation with the standard of reference) for each of the aneurysm components as compared with the composite picture. An overall score for each technique was also obtained. RESULTS: Of the 42 aneurysms examined, 34 were small (<10 mm), six were large (10 25 mm), and two were giant (>25 mm). Three aneurysms were not detected with MRA. These were smaller than 3 mm and either in an anatomically difficult location (middle cerebral artery bifurcation) or obscured by adjacent hematoma. Two large aneurysms were depicted as undersized by IA-DSA owing to the presence of intramural thrombus shown by MRA axial source data. IA-DSA received the highest scores overall and in three of the four subgroups. Three-dimensional isosurface reconstructions scored higher than did IA-DSA for depiction of the aneurysm neck, although this difference was not significant. The MPR and 3D-isosurface images were comparable to those of IA-DSA in all categories. MPR images were particularly useful for defining branch vessels and the aneurysm neck. MIP images scored poorly in all subgroups (P < .005) compared with IA-DSA findings, except for in aneurysm detection. Source data images were significantly inferior to those of IA-DSA in all categories (P < .005). CONCLUSION: MRA is currently inferior to IA-DSA in pretreatment assessment of intracranial aneurysms, and can miss small lesions (<3 mm). It can, however, provide complementary information to IA-DSA, particularly in anatomically complex areas or in the presence of intramural thrombus. If MRA is used in aneurysm assessment, a meticulous technique with reference to both axial source data and MPR is mandatory. The axial source data should not be interpreted in isolation. Three-dimensional isosurface images are comparable to those of IA-DSA and are more reliable than are MIP images, which should be interpreted with caution. PMID- 11039341 TI - Whole-brain functional MR imaging activation from a finger-tapping task examined with independent component analysis. AB - BACKGROUND AND PURPOSE: Independent component analysis (ICA), unlike other methods for processing functional MR (fMR) imaging data, requires no a priori assumptions about the hemodynamic response to the task. The purpose of this study was to analyze the temporal characteristics and the spatial mapping of the independent components identified by ICA when the subject performs a finger tapping task. METHODS: Ten healthy subjects performed variations of the finger tapping task conventionally used to map the sensorimotor cortex. The scan data were processed with ICA, and the temporal configuration of the components and their spatial localizations were studied. The locations with activation were tabulated and compared with locations known to be involved in the organization of motor functions in the brain. RESULTS: Components were identified that correlated to varying degrees with the conventional boxcar reference function. One or more of these components mapped to the sensorimotor cortex, supplementary motor area (SMA), putamen, and thalamus. By means of ICA components, sensorimotor cortex, supplementary motor area, and superior cerebellar activation were identified bilaterally in 100% of the subjects; thalamus activation was contralateral to the active hand in 80%; and putamen activation was contralateral to the active hand in 60%. CONCLUSION: ICA processing of multislice fMR imaging data acquired during finger tapping identifies the sensorimotor cortex, SMA, cerebellar, putamen, and thalamic activation. ICA appears to be a method that provides information on both the temporal and spatial characteristics of activation. Multiple task-related components can be identified by ICA, and specific activation maps can be derived from each separate component. PMID- 11039342 TI - Mapping functionally related regions of brain with functional connectivity MR imaging. AB - BACKGROUND AND PURPOSE: In subjects who are performing no prescribed cognitive task, functional connectivity mapped with MR imaging (fcMRI) shows regions with synchronous fluctuations of cerebral blood flow. When specific tasks are performed, functional MR imaging (fMRI) can map locations in which regional cerebral blood flow increases synchronously with the performance of the task. We tested the hypothesis that fcMRI maps, based on the synchrony of low-frequency blood flow fluctuations, identify brain regions that show activation on fMRI maps of sensorimotor, visual, language, and auditory tasks. METHODS: In four volunteers, task-activation fMRI and functional connectivity (resting-state) fcMRI data were acquired. A small region of interest (in an area that showed maximal task activation) was chosen, and the correlation coefficient of the corresponding resting-state signal with the signal of all other voxels in the resting data set was calculated. The correlation coefficient was decomposed into frequency components and its distribution determined for each fcMRI map. The fcMRI maps were compared with the fMRI maps. RESULTS: For each task, fcMRI maps based on one to four seed voxel(s) produced clusters of voxels in regions of eloquent cortex. For each fMRI map a closely corresponding fcMRI map was obtained. The frequencies that predominated in the cross-correlation coefficients for the functionally related regions were below 0.1 Hz. CONCLUSION: Functionally related brain regions can be identified by means of their synchronous slow fluctuations in signal intensity. Such blood flow synchrony can be detected in sensorimotor areas, expressive and receptive language regions, and the visual cortex by fcMRI. Regions identified by the slow synchronous fluctuations are similar to those activated by motor, language, or visual tasks. PMID- 11039343 TI - Correlation of myo-inositol levels and grading of cerebral astrocytomas. AB - BACKGROUND AND PURPOSE: In a limited number of patients, the level of myo inositol (MI), as seen by proton magnetic resonance spectroscopy (HMRS), has been shown to differ for gliomas of different histologic grades. We sought to determine if MI levels correlate with cerebral astrocytoma grade. METHODS: Five control subjects, 14 patients with low-grade astrocytoma, 10 patients with anaplastic astrocytoma, and 10 patients with glioblastoma multiforme (GBM) underwent single-volume HMRS with an echo time of 20 ms. Twenty-five patients had received surgery, chemotherapy, and/or radiation therapy previously. Using the curve-fitting program supplied by the manufacturer, peak areas for n-acetyl aspartate (NAA), choline (Cho), and MI were normalized with respect to the peak area of creatine (Cr). Ratios for MI/Cr, Cho/Cr, and NAA/Cr were obtained for each lesion and retrospectively compared with the histologic grade of the lesion. RESULTS: Levels of MI/Cr were higher (0.82 +/- 0.25) in patients with low-grade astrocytoma, intermediate (0.49 +/- 0.07) in control subjects, and lower in patients with anaplastic astrocytoma (0.33 +/- 0.16) and GBM (0.15 +/- 0.12). CONCLUSION: Our study shows a trend toward lower MI levels in the presence of anaplastic astrocytomas and GBMs compared with those of low-grade astrocytomas. MI levels may have implications in the grading of cerebral astrocytomas. PMID- 11039344 TI - Choroid plexus changes after temporal lobectomy. AB - BACKGROUND AND PURPOSE: Postoperative contrast-enhanced MR imaging of the brain is routinely used when evaluating for residual or recurrent brain tumor. It is imperative to be aware of morphologic changes and imaging features that typically occur in response to surgical manipulation at the postoperative site to avoid misinterpretation of imaging findings. Our purpose was to determine normal postoperative changes and alterations in the choroid plexus among patients who had undergone temporal lobectomy in order to distinguish this appearance from pathologic changes that may be seen in the presence of infection or recurrent tumors. METHODS: We reviewed 159 MR scans from 95 patients with hippocampal sclerosis or gliosis who underwent temporal lobectomy for treatment of intractable epilepsy. Choroid plexus location and size were assessed on contrast enhanced T1-weighted images. RESULTS: After temporal lobectomy, the choroid plexus enlarged and sagged into the resection site. Increase in the size of the choroid plexus occurred in 58% of cases overall. The degree of enhancement also increased after surgery, sometimes resulting in a nodular pattern of enhancement. The changes were most marked during the 1st week after temporal lobectomy, and showed an enlarged, markedly enhancing choroid plexus on 86% of the scans. CONCLUSION: Postoperative changes of the choroid plexus after temporal lobectomy include sagging into the resection site, an increased size, and an increased degree of enhancement. Normal postoperative morphologic characteristics may mimic neoplastic enhancement pattern. Familiarity with this appearance is important to avoid a pitfall in diagnosis of recurrent postoperative temporal lobe neoplasms. PMID- 11039345 TI - CT of the head by use of reduced current and kilovoltage: relationship between image quality and dose reduction. AB - BACKGROUND AND PURPOSE: CT is a frequent examination that is performed using ionizing radiation. We sought to assess image-quality changes on CT scans of the head when the radiation dose is reduced by changing tube current and kilovoltage. METHODS: A formalin-fixed cadaver was examined in conventional and helical mode by use of two CT-scanners. Surface dose was measured with standard scanning parameters, and after reduction of tube current and kilovoltage. Five experienced examiners independently evaluated subjective image quality. RESULTS: In the conventional mode, the highest surface dose was 83.2 mGy (scanner 1: helical mode, 55.6 mGy), and 66.0 mGy (scanner 2: helical mode, 55.9 mGy). By changing kVp and mAs, a dose reduction of up to 75% (scanner 1), and 60% (scanner 2) was achieved. No observable differences in image quality between scans obtained with doses from 100% to 60% of standard settings were noted. Ten of 20 images obtained with the highest dose and 13 of 20 images obtained with lowest dose (19-29.4 mGy) were reliably identified by subjective quality assessment. Scans produced with a surface dose of less than 30 mGy were judged uninterpretable. CONCLUSION: Standard parameters used in cranial CT are oriented toward best image quality. A dose reduction up to 40% may be possible without loss of diagnostic image quality. PMID- 11039346 TI - Optic tract edema in a meningioma of the tuberculum sellae. AB - We report a case of tuberculum sellae meningioma with optic tract edema. Contrary to a prior report on this topic, edema along the optic tract is not only seen in craniopharyngiomas but may be seen (although rarely) in other common parasellar tumors, as in our case of a tuberculum sellae meningioma. The pathogenesis of this edema in meningioma is controversial. PMID- 11039347 TI - MR imaging of the enlarged endolymphatic duct and sac syndrome by use of a 3D fast asymmetric spin-echo sequence: volume and signal-intensity measurement of the endolymphatic duct and sac and area measurement of the cochlear modiolus. AB - BACKGROUND AND PURPOSE: In enlarged endolymphatic duct (EED) and sac (EES) syndrome, deformity of the EED and EES is congenital; however, hearing loss is acquired. To investigate the pathophysiology of progressive sensorineural hearing loss in EED and EES syndrome, we measured the volume of the EED and EES, the diameter of the EED and EES, the area of the cochlear modiolus, and the signal intensity of the EES and compared our findings against degree of hearing loss. METHODS: Thin-section MR images of 33 ears in 17 patients with EED and EES syndrome were studied. All studies were obtained on a 1.5-T MR unit using a quadrature surface phased-array coil. Heavily T2-weighted 3D fast asymmetric spin echo images were obtained with a voxel size of 0.3 x 0.3 x 0.8 mm without zero fill interpolation. Two radiologists traced the areas of the EED and EES manually, and the volume was calculated. The area of the cochlear modiolus, diameter of the EED and EES, and signal intensity of the EES were also measured by drawing regions of interest manually. The signal intensity ratio of EES/CSF was calculated. These measured values were compared against audiographic data, and the degree of linear correlation was determined. RESULTS: The volume of the EED and EES, the area of the modiolus, the diameter of the EED and EES, and the signal intensity of the EES did not show significant correlation with degree of hearing loss. CONCLUSION: These findings suggest that there is a microscopic area of damage or fragility in the inner ear not visible even with thin-section heavily T2-weighted MR imaging. PMID- 11039348 TI - Fenestration surgery for otosclerosis: CT findings of an old surgical procedure. AB - Numerous attempts to deal surgically with otosclerosis were made before the current method of stapedectomy with stapes prosthesis was established. We report a case with unique CT findings of a patient who underwent fenestration surgery for otosclerosis in the early 1940s. Recognition of this old surgical procedure on the imaging scans may avoid misdiagnosis of labyrinthine fistulae or middle and inner ear malformations. PMID- 11039349 TI - Neurosyphilis as a cause of facial and vestibulocochlear nerve dysfunction: MR imaging features. AB - The prevalence of syphilis increased for several decades before the mid-1990s in the United States, particularly in the southern states. We report a case of neurosyphilis causing bilateral facial and vestibulocochlear nerve dysfunction in which the diagnosis was not initially suspected based on the patient's demographics and history. The MR imaging features helped to make the diagnosis in this case and to exclude other possible causes of multiple cranial nerve dysfunction in this patient. Hearing loss associated with neurosyphilis is one of the few treatable forms of progressive hearing loss, and it is essential that a diagnosis of neurosyphilis be made expeditiously. PMID- 11039350 TI - Traumatic neuroma after neck dissection: CT characteristics in four cases. AB - BACKGROUND AND PURPOSE: Traumatic neuroma, an attempt by an injured nerve to regenerate, may present as a palpable nodule or an area sensitive to touch (trigger point) after neck dissection. The purpose of this study was to identify CT characteristics of traumatic neuroma in four patients after neck dissection. METHODS: Between April 1995 and November 1998, the CT studies in three men and one woman (ages, 45-64 years) who had had a radical neck dissection and a nodule posterior to the carotid artery were reviewed retrospectively. CT was performed 1.5 to 6 years after neck dissection with clinical correlation and/or pathologic examination. Three patients had squamous cell carcinoma of the upper aerodigestive tract and one had a primary parotid adenocarcinoma. RESULTS: Three patients with a traumatic neuroma had a centrally radiolucent nodule with peripherally dense rim and intact layer of overlying fat, which was stable on CT studies for 1 to 2 years. One of these had a clinical trigger point. The fourth patient with a pathologically proved traumatic neuroma mixed with tumor had intact overlying fat, but the nodule lacked a radiolucent center and was not close to the carotid artery. CONCLUSION: The CT findings of a stable nodule that is posterior but close to the carotid artery with central radiolucency, a dense rim, and intact overlying fat, combined with the clinical features of a trigger point and a lack of interval growth, strongly suggest the diagnosis of traumatic neuroma. PMID- 11039351 TI - MR imaging for predicting neoplastic invasion of the cervical esophagus. AB - BACKGROUND AND PURPOSE: Esophageal invasion (EI) by head and neck neoplasm has important prognostic and surgical management implications. Our purpose was to determine the accuracy of MR imaging for predicting neoplastic cervical esophageal invasion. METHODS: MR scans of the neck obtained from 22 patients with periesophageal masses were retrospectively reviewed independently and by consensus by two experienced head and neck radiologists who were unaware of surgical findings. The patients were selected from clinical, radiologic, or pathologic reports suggesting EI. The following imaging criteria for EI were evaluated: effacement of periesophageal fat planes, circumferential mass, paraesophageal lymph nodes, luminal size, wall thickening, increased T2 wall signal, and wall enhancement. There were eight patients with EI and 14 patients without EI, as confirmed by surgical findings or pathologic examination. RESULTS: The consensus criteria with the best sensitivities were any wall thickening (100%), effaced fat plane (100%), and any T2 wall signal abnormality (100%). The criteria with the best specificities were circumferential mass greater than 270 (100%) or 180 degrees (93%) and focal T2 wall signal abnormality (86%). The overall kappa value for the two readers for all criteria was 0.57 (moderate agreement). CONCLUSION: A circumferential mass or focal T2 signal abnormality on the esophageal wall suggests the presence of EI. An intact fat plane, absence of wall thickening, and no T2 wall signal abnormalities imply that the esophagus is not invaded. PMID- 11039352 TI - Fast MR imaging of fetal CNS anomalies in utero. AB - BACKGROUND AND PURPOSE: Although sonography is the primary imaging technique for evaluating the developing fetus, significant limitations exist in the sonographic prenatal diagnosis of many brain disorders. Fast MR imaging is increasingly being used to determine the underlying cause of nonspecific fetal CNS abnormalities detected sonographically and to confirm or provide further support for such anomalies. Our goal was to determine the value of MR imaging in establishing the diagnosis of fetal CNS anomalies, to ascertain how this information might be used for patient counseling, and to assess its impact on pregnancy management. METHODS: We prospectively performed MR examinations of 73 fetuses (66 pregnancies) with suspected CNS abnormalities and compared these with available fetal sonograms, postnatal images, and clinical examinations. Retrospectively, the impact on patient counseling and pregnancy management was analyzed. RESULTS: Images of diagnostic quality were routinely obtained with in utero MR imaging, which was particularly valuable in detecting heterotopia, callosal anomalies, and posterior fossa malformations, and for providing excellent anatomic information. We believe that 24 (46%) of 52 clinical cases were managed differently from the way they would have been on the basis of sonographic findings alone. In every case, the referring physicians thought that MR imaging provided a measure of confidence that was not previously available and that was valuable for counseling patients and for making more informed decisions. CONCLUSION: Sonography is the leading technique for fetal assessment and provides reliable, inexpensive diagnostic images. Fast MR imaging is an important adjunctive tool for prenatal imaging in those instances in which a complex anomaly is suspected by sonography, when fetal surgery is contemplated, or when a definitive diagnosis cannot be determined. PMID- 11039353 TI - MR, CT, and plain film imaging of the developing skull base in fetal specimens. AB - BACKGROUND AND PURPOSE: The developing fetal skull base has previously been studied via dissection and low-resolution CT. Most of the central skull base develops from endochondral ossification through an intermediary chondrocranium. We traced the development of the normal fetal skull base by using plain radiography, MR imaging, and CT. METHODS: Twenty-nine formalin-fixed fetal specimens ranging from 9 to 24 weeks' gestational age were examined with mammographic plain radiography, CT, and MR imaging. Skull base development and ossification were assessed. RESULTS: The postsphenoid cartilages enclose the pituitary and fuse to form the basisphenoid, from which the sella turcica and the posterior body of the sphenoid bone originate. The presphenoid cartilages will form the anterior body of the sphenoid bone. Portions of the presphenoid cartilage give rise to the mesethmoid cartilage, which forms the central portion of the anterior skull base. Ossification begins in the occipital bone (12 weeks) and progresses anteriorly. The postsphenoid (14 weeks) and then the presphenoid portion (17 weeks) of the sphenoid bone ossify. Ossification is seen laterally (16 weeks) in the orbitosphenoid, which contributes to the lesser wing of the sphenoid, and the alisphenoid (15 weeks), which forms the greater wing. CONCLUSION: MR imaging can show early progressive ossification of the cartilaginous skull base and its relation to intracranial structures. The study of fetal developmental anatomy may lead to a better understanding of abnormalities of the skull base. PMID- 11039354 TI - Prominent basal emissary foramina in syndromic craniosynostosis: correlation with phenotypic and molecular diagnoses. AB - BACKGROUND AND PURPOSE: Jugular foraminal stenosis (JFS) or atresia (JFA) with collateral emissary veins (EV) has been documented in syndromic craniosynostosis. Disruption of EV during surgery can produce massive hemorrhage. Our purpose was to describe the prevalence of prominent basal emissary foramina (EF), which transmit enlarged EV, in syndromic craniosynostosis. Our findings were correlated with phenotypic and molecular diagnoses. METHODS: We reviewed the medical records and imaging examinations of 33 patients with syndromic craniosynostosis and known fibroblast growth factor receptor (FGFR) mutations. All patients underwent CT and 14 MR imaging. The cranial base was assessed for size of occipitomastoid EF and jugular foramina (JF). Vascular imaging studies were available from 12 patients. A control group (n = 76) was used to establish normal size criteria for JF and EF. RESULTS: Phenotypic classification included Crouzon syndrome (n = 10), crouzonoid features with acanthosis nigricans (n = 3), Apert syndrome (n = 10), Pfeiffer syndrome (n = 4), and clinically unclassifiable bilateral coronal synostosis (n = 6). EF > or = 3 mm in diameter and JFS or JFA were identified in 23 patients with various molecular diagnoses. Vascular imaging in patients with JFS or JFA and enlarged EF revealed atresia or stenosis of the jugular veins and enlarged basal EV. JFA was seen in all patients with the FGFR3 mutation with crouzonoid features and acanthosis nigricans. Four patients had prominent EF without JFS. Six patients had normal JF and lacked enlarged EF. CONCLUSION: Enlarged basal EF are common in syndromic craniosynostosis and are usually associated with JFS or JFA. Bilateral basilar venous atresia is most common in patients with the FGFR3 ala391glu mutation and crouzonoid features with acanthosis nigricans, but may be found in patients with FGFR2 mutations. Skull base vascular imaging should be obtained in patients with syndromic craniosynostosis with enlarged EF. PMID- 11039355 TI - Parietal lipoma associated with cortical dysplasia and abnormal vasculature: case report and review of the literature. AB - We present the case of an unusually located intracranial lipoma in a 17-year-old patient with partial epilepsy who was being controlled with medication. The lipoma was located deep in the left sylvian fissure, in the inferior parietal lobule, associated with cortical dysplasia of the surrounding supramarginal gyrus. Abnormal vasculature was detected adjacent to and within the adipose mass. The findings of the imaging studies that included CT, MR imaging, and MR angiography, are described along with a brief review of the literature. PMID- 11039356 TI - Modification of a previously described arteriovenous malformation model in the swine: endovascular and combined surgical/endovascular construction and hemodynamics. AB - BACKGROUND AND PURPOSE: The rete mirabile in swine has been proposed as an arteriovenous malformation (AVM) model for acute experimental studies through surgical creation of a large carotid-jugular fistula. This report describes two endovascular modifications to simplify the surgical creation and provides hemodynamic parameters for the AVM model. METHODS: An AVM model was created in 29 animals to study n-butyl 2-cyanoacrylate polymerization kinetics. The common carotid artery (CCA) was punctured and a guiding catheter was inserted tightly into the origin of the ascending pharyngeal artery (APA). The CCA was ligated proximal to the catheter to create a pressure drop across the rete, which represented the AVM nidus. The catheter hub was opened whenever needed and served as the venous drainage of the AVM nidus. The contralateral APA served as the arterial feeder. Instead of the surgical ligation of the CCA, a temporary balloon occlusion was performed in three animals. RESULTS: A mean pressure gradient of 14.9 +/- 10.5 mm Hg (range, 4-42 mm Hg) was measured across the rete. The mean flow rate was 30.4 +/- 14.2 mL/min (range, 3.5-46 mL/min), as measured at the venous drainage. CONCLUSION: The endovascular and combined surgical-endovascular rete AVM model in swine is easy to construct and is less time-consuming than are the currently used models for acute experimental studies. Hemodynamic parameters can be monitored during the entire experiment and correspond to values found in human cerebral AVMs. PMID- 11039357 TI - Endovascular treatment of experimental aneurysms by use of a combination of liquid embolic agents and protective devices. AB - BACKGROUND AND PURPOSE: The use of liquid embolic agents for embolization of cerebral aneurysms has been reported in the neurosurgical literature. The most important limitation of this technique is the relatively poor control of migration of the liquid embolic agent into the parent artery. We performed an experimental aneurysm study using a liquid embolic agent and different protective devices to evaluate the safety and technical feasibility of this endovascular technique. METHODS: Forty lateral aneurysms were surgically constructed on 20 common carotid arteries of swine. Onyx alone was used to obliterate eight aneurysms. Onyx was also used in combination with microcoils (n = 11), microstents (n = 6), balloons inflated proximally to the neck of the aneurysm (n = 6), and across the neck of the aneurysm (n = 7). One control aneurysm was embolized with Guglielmi detachable coils (GDCs) alone. RESULTS: The use of a microballoon across the neck of the aneurysm, a microstent deployed across the neck of the aneurysm, or the deposit of GDCs into the aneurysm allowed faster and more complete filling of the aneurysm with Onyx. However, these protection devices did not totally preclude intractable migration of Onyx into the parent artery (migration rate, 9-33%). CONCLUSION: Although complete occlusion of experimental aneurysms with Onyx is feasible using protective devices, migration of the liquid embolic agent into the parent artery or intracranially remains a difficult challenge. Further experimental studies need to be performed to master this technique and to select those aneurysms that can be safely treated in clinical practice. PMID- 11039358 TI - Carotid artery stenting: technical considerations. AB - BACKGROUND AND PURPOSE: Carotid endarterectomy (CEA) is one of the most frequently performed operations in the United States. To offer patients a less invasive means to achieve the same goal, carotid artery stenting (CAS) is investigated as an alternative treatment to CEA. METHODS: Three hundred ninety patients underwent CAS, with 451 vessels treated. CAS was performed using a coaxial system with a 7F 90-cm sheath for predilation, stent placement, and stent dilation. Pretreatment antiplatelet therapy was administered. We currently practice same-day admissions and 23-hour discharges. RESULTS: The technical success rate was 98%. The 30-day mortality/morbidity rates were as follows: death, 1.7% (two [0.5%] neurologic and five [1.2%] systemic] major strokes, 0.9% (two of four were related to the intervention); minor strokes, 5.5%. Among 25 patients who suffered minor strokes, 14 achieved complete recovery. On an annual basis, the incidence of minor stroke declined from 6.8% (1994-1995), to 5.8% (1995-1996), 5.3% (1996-1997), and then 4% (1997-1998), with no major strokes or neurologic deaths occurring during the 1997 to 1998 period. CONCLUSION: CAS is an effective treatment for carotid stenosis. With proper selection of patients and meticulous technique, complication rates compare favorably with those of CEA. PMID- 11039359 TI - Primary stenting for high-grade basilar artery stenosis. AB - We report two patients with symptomatic high-grade stenosis of the basilar artery refractory to appropriate maximal medical therapy in whom endovascular stenting was performed successfully without preliminary balloon angioplasty. Excellent angiographic results were achieved and there were no procedural or periprocedural complications. The patients were asymptomatic and neurologically intact at a mean clinical follow-up of 6.5 months. Primary stenting of basilar artery stenosis may be an alternative to balloon angioplasty for patients with symptomatic lesions refractory to medical therapy or in whom anticoagulation is contraindicated. PMID- 11039360 TI - Abciximab rescue in acute carotid stent thrombosis. AB - Occlusion of an internal carotid artery stent was identified immediately post placement in a patient who had restenosis after prior angioplasty. An IV dose of abciximab was administered, and serial angiograms were performed. This resulted in partial resolution of the thrombus at 10 minutes and complete resolution at 20 minutes. PMID- 11039361 TI - Double-balloon technique for embolization of carotid cavernous fistulas. AB - Embolization of a carotid cavernous fistula (CCF) by means of a detachable balloon is an established method for treating CCFs while preserving a patent parent internal carotid artery (ICA). However, failure to embolize the CCF may occur on a few occasions, such as when the balloon cannot pass through the fistula into the cavernous sinus by blood flow, or when the inflated balloon in the cavernous sinus retracts to the carotid artery. Under these circumstances, the ICA may have to be sacrificed in order to treat the CCF. Herein we describe a double-balloon technique for embolization of a CCF. By applying this technique, we successfully treated nine of 11 CCFs, without compromise of the parent ICA when the conventional one-balloon technique failed. PMID- 11039362 TI - Postoperative evaluation for disseminated medulloblastoma involving the spine: contrast-enhanced MR findings, CSF cytologic analysis, timing of disease occurrence, and patient outcomes. AB - BACKGROUND AND PURPOSE: Postoperative MR imaging is routinely performed for staging of medulloblastoma because of frequent tumor dissemination along CSF pathways. The goals of this study were to: 1) determine the timing of disease occurrence and contrast-enhanced MR imaging features of disseminated medulloblastoma involving the spine and their relationship to patient outcomes; and 2) compare the diagnostic accuracy of MR imaging findings with CSF cytologic analysis. METHODS: Medical records, pathologic reports, and unenhanced and contrast-enhanced postoperative MR images of the spine and head from 112 patients who had resection of medulloblastoma were retrospectively reviewed. MR images of the spine were evaluated for abnormal contrast enhancement in the meninges and vertebral bone marrow. MR images of the head were evaluated for recurrent or residual intracranial tumor. Imaging data were correlated with available CSF cytologic results and patient outcomes. RESULTS: Twelve patients (11%) had tumor within the spinal leptomeninges depicted on MR images at the time of diagnosis. Twenty-five patients (22%) had disseminated disease in the spine (leptomeninges, n = 22; vertebral marrow, n = 1; or both locations, n = 2) on MR images 2 months to 5.5 years (mean, 2 years) after initial surgery and earlier negative imaging examinations. Eleven other patients (10%) had recurrent intracranial medulloblastoma without spinal involvement seen with MR imaging. Spinal MR imaging had a sensitivity of 83% in the detection of disseminated tumor, whereas contemporaneous CSF cytologic analysis had a sensitivity of 60%. The sensitivity of CSF cytologic analysis increased to 78% with acquisition of multiple subsequent samples, although diagnosis would have been delayed by more than 6 months compared with diagnosis by spinal MR imaging in six patients. Spinal MR imaging was found to have greater overall diagnostic accuracy than CSF cytologic analysis in the early detection of disseminated tumor (P = .03). Spinal MR imaging confirmed disseminated tumor when contemporaneous CSF cytologic findings were negative in 13 patients, whereas the opposite situation occurred in only two patients. False-positive results for spinal MR imaging and CSF cytologic analysis occurred when these examinations were obtained earlier than 2 weeks after surgery. The 5-year survival probability for patients with spinal tumor was 0.24 +/- 0.08 versus 0.68 +/- 0.05 for the entire study group. CONCLUSION: Spinal MR imaging was found to have greater diagnostic accuracy than CSF cytologic analysis in the early detection of disseminated medulloblastoma. CSF cytologic analysis infrequently confirmed disseminated tumor when spinal MR imaging results were negative. Delaying spinal MR imaging and CSF cytologic analysis by more than 2 weeks after surgery can reduce false-positive results for both methods. The presence of disseminated medulloblastoma in the spine seen with MR imaging is associated with a poor prognosis. PMID- 11039363 TI - MR findings of the brain stem in arterial hypertension. PMID- 11039364 TI - Imaging macrophage activity in the brain by using ultrasmall particles of iron oxide. PMID- 11039365 TI - In re: histological and morphologic comparison of experimental aneurysms with human intracranial aneurysms. PMID- 11039366 TI - Predictive value of computed tomographic myelography in obstetrical brachial plexus palsy. AB - Preoperative radiologic studies to detect root avulsions of the brachial plexus caused by birth trauma are considered useful in assisting with surgical planning for reconstruction. In this study, the predictive value of computed tomographic (CT) myelography in detecting nerve root avulsions at our institution was determined. Sixty-three consecutive patients with an obstetrical brachial plexus palsy who had had both preoperative CT myelography and reconstructive surgery were selected. All CT myelograms were analyzed post hoc by a single neuroradiologist in a manner blind to the surgical findings. At each root level of the brachial plexus, the presence of a pseudomeningocele was noted along with the presence or absence of rootlets within each identified pseudomeningocele. Extraforaminal root avulsions later determined at surgery were reviewed by a single surgeon in a manner blind to the radiographic results. Surgical and radiographic findings were then compared at each corresponding root level. A total of 281 roots were examined. The sensitivity, specificity, positive predictive value, and likelihood ratio for root avulsions with pseudomeningoceles were 0.63, 0.85, 0.40, and 4.2, respectively. For pseudomeningoceles for which rootlets traversing the sac could not be identified, these values were 0.37, 0.98, 0.74, and 18.5, respectively. The presence of pseudomeningoceles with or without rootlets was not a sensitive indicator of root avulsions. Root avulsions were better predicted by identifying the absence of rootlets in a pseudomeningocele. This absence on CT myelography may be used to suggest an extraforaminal root avulsion due to its high specificity and high likelihood ratio. PMID- 11039367 TI - Formation of philtral column using vertical interdigitation of orbicularis oris muscle flaps in secondary cleft lip. AB - The philtrum in the lip has an important aesthetic significance and is a mark of individual distinction. For patients who have undergone cleft lip surgery, the construction of the philtrum is crucial for restoring a normal appearance to the upper lip. A total of 13 patients with unilateral cleft lip nose deformities were treated for the creation of a philtral column between January of 1998 and February of 1999. Eight of the patients were male and five were female with an age range of 10 to 40 years old. The scar on the philtral column is excised and a full-thickness incision is made down to the orbicularis oris muscle and mucosa. The medial and lateral muscle flaps are then exposed and split into two leaves. The two leaves of each muscle flap are sutured together to create a vertical interdigitation. Any excess skin is not excised but rather closed with 7-0 nylon. The follow-up period ranged from 6 to 15 months, with an average of 10 months. Ten of 13 patients were satisfied with their good surgical results. Two had fair results. One patient experienced a widening of the scar and no improvement in the philtral column. A possible cause for this lack of improvement was a partial disruption of the interdigitated muscle flaps due to the early active movement of the muscle before wound healing. In conclusion, the advantages of this procedure include the creation of an anatomically natural philtrum through preserving the continuity and function of the muscle, sufficient augmentation of the philtral column by the vertical interdigitation of the muscle, relief of skin tension, and no donor-site morbidity. PMID- 11039368 TI - Dog bites in New York City. AB - The purpose of this review is to characterize the public health problem of dog bites in New York City. Dog bites represent a major source of morbidity, mortality, disability, and health care cost in the United States. The most severe injuries are frequently referred to the plastic surgeon. The authors have recently treated several severe cases of unprovoked dog attacks in children. To characterize the dog bite problem in New York City, data were obtained for 1998 from the New York City Department of Health. During that year, 6568 bites were reported by the mandated physician reporters in the five boroughs of New York City. The upper extremities, lower extremities, and face were predominantly affected. The peak incidence occurred during the summer months and in children ages 7 to 9 years old. Preventive measures are discussed. PMID- 11039369 TI - Reduction mammaplasty provides long-term improvement in health status and quality of life. AB - The objective of this study was to assess health status and quality of life in macromastia patients undergoing reduction mammaplasty. From January of 1997 to June of 1997, the Department of Reconstructive Plastic Surgery, at Stockholm Soder Hospital/Karolinska Hospital, conducted a prospective questionnaire study with preoperative and postoperative (6 and 12 months) assessments in 49 women who were 20 years or older. The questionnaire included four parts: Part I assessed pain (scale 1 to 10) in the neck, shoulders, back, breast, bra strap indention, and head. Part II assessed effects of breast size and weight on body posture, sleep, choice of clothing, sexual relations, and working capacity (scale 1 to 10). Part III assessed preoperative expectations for the operation in comparison with postoperative result (scale 1 to 6). Part TV included SF-36, an international health-related quality-of-life questionnaire, which has been standardized for Swedish women. As a result, reduction mammaplasty (mean resection weight, 1052 g) provided significant reduction of pain in all locations (p < 0.001). The improvements continued up to 12 months postoperatively. The patients' main subjective problems related to the size and weight of the breast were body posture and choice of clothing. The patients scored significant improvements of all subjective problems (p < 0.001), except sleep. The patients' expectations were met to a high extent. In some areas such as intimate situations, femininity, and social contacts, the results exceeded the preoperative expectations. Preoperatively, the mammaplasty patients scored significantly lower (p < 0.05 to p < 0.001, depending on area) in SF-36, i.e., the patients had lower quality of life compared with women in the same age group. Reduction mammaplasty resulted in significantly improved quality of life; furthermore, the results were similar after 6 and 12 months, indicating long-term improvement. In fact, after 1 year, there was no statistically significant difference between the patients who had been operated on and the age-matched women, i.e., the women were normalized in health-related quality of life as judged by the SF-36. PMID- 11039370 TI - Relationship of obesity and specimen weight to complications in reduction mammaplasty. AB - Obesity and specimen weight have both been associated with a higher incidence of complications for patients undergoing reduction mammaplasty. However, obesity has been arbitrarily and inconsistently defined, and the degree of obesity has not been considered in these previous studies. Because insurance companies are increasingly demanding weight loss before authorizing surgery, the relationship of obesity and breast size to complications is of great importance. Upon critical review of the literature, a number of fundamental questions remain unanswered. If complications are more frequent in the obese patient, are these complications directly proportional to the degree of obesity? Also, if the patient is required to lose weight before surgery, is weight loss effective in reducing complication rates? In an attempt to clarify these issues, 395 patients who underwent reduction mammaplasty over a 10-year period were reviewed retrospectively. Patients were arbitrarily divided into five groups in which, depending on their degree of relative obesity, they were classified as less than 5 percent, 5 to 10 percent, 10 to 15 percent, 15 to 20 percent, or greater than 20 percent above average body weight. To evaluate the relationship of specimen weight to complications, patients were also arbitrarily divided into five groups according to weight of the breast reduction specimen, which was classified as less than 300 g, 300 to 600 g, 600 to 900 g, 900 to 1200 g, and greater than 1200 g reduction per breast. Complications were then divided into local and systemic and major and minor. When bilateral reductions alone were analyzed (n = 267), there was a statistically significant increase in complication rate in the obese (p = 0.01). However, when the obese population was further subdivided according to their degree of obesity (less than 5 percent, 5 to 10 percent, 10 to 15 percent, 15 to 20 percent, and greater than 20 percent above average body weight), no further correlation was found. However, the relationship between specimen weight per breast and complications was much stronger with a direct correlation existing between increasing specimen weight and the incidence of complications. Although this study has shown that patients who are average body weight have fewer complications than obese patients after breast reduction surgery, it has not shown an increasing incidence of complication with increasing degrees of obesity. The implications of these findings and their relationship for denying patients surgery on the basis of weight alone are discussed in detail. PMID- 11039371 TI - Secondary reduction mammaplasty: is using a different pedicle safe? AB - Reduction mammaplasty is a frequently performed procedure and one with consistent patient satisfaction. Few patients present for revisional procedures, and even fewer present for a secondary or repeated reduction mammaplasty. This study defines secondary reduction mammaplasty as performing an additional reduction using a pedicled nipple-areola complex. Few reports of secondary reduction are found in the literature. Operative guidelines for secondary reduction mammaplasty have been published recently. However, the experience of others has differed from these guidelines, and herein is presented another experience with secondary reduction mammaplasty. Ten cases of secondary reduction over a 37-year period were identified and reviewed. The initial reductions were performed using six different techniques. An average of 307 g of tissue per breast (range, 130 to 552 g) was removed at the initial operations. The secondary reductions were performed using four different techniques, and an average of 458 g of tissue per breast (range, 147 to 700 g) was removed at the secondary operations. Three of the 10 patients underwent initial and secondary reduction with the same technique. An average of 4 years (range, 1 to 10 years) separated these surgeries. Seven of the 10 patients underwent initial and secondary reductions with different technique. An average of 15 years (range, 5 to 19 years) separated these procedures. There was an average 5-year follow-up (range, 1 to 20 years) in this series. Four of the 10 patients experienced self-limiting complications after secondary reduction, including delay in wound healing, delay in the return of nipple sensitivity, and mild fat necrosis. Three of the four patients with complications had undergone secondary reduction with a different pedicle technique. No significant or long-lasting skin, pedicle, or nipple-areola complex compromise was found after secondary reduction mammaplasty. In contrast to the recently published guidelines, this study demonstrates that secondary reduction mammaplasty is a safe and viable option when performed with either similar or different technique. This finding allows secondary reduction mammaplasty to be tailored to the individual breast type and to the abilities of the specific surgeon. PMID- 11039372 TI - Estrogen and progesterone receptors in gynecomastia. AB - The etiology of gynecomastia is unknown. There seems to be no increased incidence of malignancies in patients with idiopathic gynecomastia; however, patients with Klinefelter syndrome exhibit an increased incidence of malignancy. The authors reviewed the results of 34 patients with gynecomastia diagnosed in adolescence who, following initial evaluation, had a mastectomy. The estrogen and progesterone receptors were analyzed in these patients. Three of the patients were diagnosed with Klinefelter syndrome. These three patients exhibited elevated amounts of estrogen and progesterone receptors. None of the patients who were not diagnosed with this syndrome demonstrated significant elevation of their estrogen or progesterone receptors. The presence of elevated estrogen and progesterone receptors in patients with Klinefelter syndrome provides a potential mechanism by which these patients may develop breast neoplasms. The absence of elevated estrogen and progesterone receptors in patients with idiopathic gynecomastia may serve to clarify why these patients' disease rarely degenerates into malignancy. PMID- 11039373 TI - Prospective analysis of psychosocial outcomes in breast reconstruction: one-year postoperative results from the Michigan Breast Reconstruction Outcome Study. AB - In the past decade, changing attitudes toward breast reconstruction among both patients and providers have led a growing number of women to seek breast reconstruction after mastectomy. Although investigators have documented the psychological, social, emotional, and functional benefits of breast reconstruction, little research has evaluated the effects of procedure choice on these outcomes. The current study prospectively evaluated and compared psychosocial outcomes for three common options for mastectomy reconstruction: tissue expander/implant, pedicle TRAM, and free TRAM techniques. In a prospective cohort design, patients undergoing postmastectomy reconstruction for the first time with expander/implant, pedicle TRAM, or free TRAM procedures were recruited from 12 centers and 23 plastic surgeons in the United States and Canada. Before reconstruction and at 1 year after reconstruction, patients were evaluated by a battery of questionnaires consisting of both generic and condition-specific surveys. Outcomes assessed included emotional well-being, vitality, general mental health, social functioning, functional well-being, social well-being, and body image. Baseline (preoperative) scores and the change in scores (the difference between postoperative and preoperative scores) were compared across procedure types using t tests and analysis of covariance. Preoperative and 1-year postoperative surveys were obtained from 273 patients. Procedure type was reported in 250 patients, of whom 56 received implant reconstructions, 128 pedicle TRAM flaps, and 66 free TRAM flaps. A total of 161 immediate and 89 delayed reconstructions were performed. Among women receiving immediate reconstruction, significant improvements were observed in all psychosocial variables except body image. However, no significant effects of procedure type on these changes over time existed. Similarly, delayed reconstruction patients had significant increases in emotional well-being, vitality, general mental health, functional well-being, and body image. Although the choice of reconstructive technique did not significantly impact most of these outcomes, significant differences existed among procedure types for three psychosocial subscales. Patients undergoing delayed expander/implant reconstructions reported greater improvements in vitality and social well-being relative to women receiving delayed TRAM procedures. By contrast, delayed TRAM patients noted significantly greater gains in body image compared with women choosing delayed expander-implant reconstruction. The authors conclude that both immediate and delayed breast reconstructions provide substantial psychosocial benefits for mastectomy patients. Although the choice of reconstructive procedure does not seem to significantly affect improvements in psychosocial status with immediate reconstruction, our data suggest that procedure type does have a significant effect on gains in vitality and body image for women undergoing delayed reconstruction. PMID- 11039374 TI - Postoperative complications and functional results after total glossectomy with microvascular reconstruction. AB - Microsurgical reconstruction after total glossectomy can greatly improve quality of life; however, postoperative functional results are often unstable, and the effectiveness of total glossectomy remains questionable. To determine the problems of reconstruction after total glossectomy with laryngeal preservation and to examine the functional results of swallowing and speech, 30 patients who had undergone total glossectomy and reconstruction with free flaps were reviewed for this study. The patients ranged in age from 20 to 73 years, and 23 of the 30 had undergone reconstruction with a rectus abdominis musculocutaneous flap. Wider and thicker flaps were designed and transferred and were sutured to suspend the larynx. To maintain physiologic swallowing function after surgery, the extent of laryngeal suspension and cricopharyngeal myotomy was limited. Of the 30 patients, 21 (70 percent) could be decannulated with laryngeal preservation; 20 of these 21 could tolerate a normal/soft/pureed diet, and 1 was limited to a fluid diet. Speech was intelligible in 16 of the 19 patients evaluated. In 9 of the 30 patients, laryngeal function could not be preserved. In four of these nine patients, additional resection combined with total glossectomy caused severe aspiration and recurrent pneumonia. Two patients with preoperative cerebral dysfunction were also poor candidates for laryngeal preservation. Additionally, the transferred flap's lack of bulk in the oral cavity and the advanced age (73 years) of one patient and the poor motivation of another may have contributed to postoperative aspiration. Aspiration occurred in one patient because of local recurrence of a tumor. The presence of preoperative cerebral dysfunction (p = 0.025), resection of the epiglottis (p = 0.005), and postoperative orocutaneous fistulas (p = 0.04) were significantly associated with the failure of laryngeal preservation. However, because of the difficulty of enrolling a sufficient number of patients in the study and the inherent limitations of retrospective studies, multivariate analysis in this study showed that no factors, such as patient age, flap volume, and the type of neck dissection, were significant predictors of laryngeal preservation. Although prospective studies are necessary, the function of individual patients must be assessed so that the study experiences discussed here can be applied to subsequent patients. PMID- 11039375 TI - A randomized prospective study of polyglycolic acid conduits for digital nerve reconstruction in humans. AB - This article reports the first randomized prospective multicenter evaluation of a bioabsorbable conduit for nerve repair. The study enrolled 98 subjects with 136 nerve transections in the hand and prospectively randomized the repair to two groups: standard repair, either end-to-end or with a nerve graft, or repair using a polyglycolic acid conduit. Two-point discrimination was measured by a blinded observer at 3, 6, 9, and 12 months after repair. There were 56 nerves repaired in the control group and 46 nerves repaired with a conduit available for follow-up. Three patients had a partial conduit extrusion as a result of loss of the initially crushed skin flap. The overall results showed no significant difference between the two groups as a whole. In the control group, excellent results were obtained in 43 percent of repairs, good results in 43 percent, and poor results in 14 percent. In those nerves repaired with a conduit, excellent results were obtained in 44 percent, good results in 30 percent, and poor results in 26 percent (p = 0.46). When the sensory recovery was examined with regard to length of nerve gap, however, nerves with gaps of 4 mm or less had better sensation when repaired with a conduit; the mean moving two-point discrimination was 3.7 +/- 1.4 mm for polyglycolic acid tube repair and 6.1 +/- 3.3 mm for end-to-end repairs (p = 0.03). All injured nerves with deficits of 8 mm or greater were reconstructed with either a nerve graft or a conduit. This subgroup also demonstrated a significant difference in favor of the polyglycolic acid tube. The mean moving two-point discrimination for the conduit was 6.8 +/- 3.8 mm, with excellent results obtained in 7 of 17 nerves, whereas the mean moving two-point discrimination for the graft repair was 12.9 +/- 2.4 mm, with excellent results obtained in none of the eight nerves (p < 0.001 and p = 0.06, respectively). This investigation demonstrates improved sensation when a conduit repair is used for nerve gaps of 4 mm or less, compared with end-to-end repair of digital nerves. Polyglycolic acid conduit repair also produces results superior to those of a nerve graft for larger nerve gaps and eliminates the donor-site morbidity associated with nerve-graft harvesting. PMID- 11039376 TI - The effects of ionizing radiation on osteoblast-like cells in vitro. AB - The well-described detrimental effects of ionizing radiation on the regeneration of bone within a fracture site include decreased osteocyte number, suppressed osteoblast activity, and diminished vascularity. However, the biologic mechanisms underlying osteoradionecrosis and the impaired fracture healing of irradiated bone remain undefined. Ionizing radiation may decrease successful osseous repair by altering cytokine expression profiles resulting from or leading to a change in the osteoblastic differentiation state. These changes may, in turn, cause alterations in osteoblast proliferation and extracellular matrix formation. The purpose of this study was to investigate the effects of ionizing radiation on the proliferation, maturation, and cytokine production of MC3T3-E1 osteoblast-like cells in vitro. Specifically, the authors examined the effects of varying doses of ionizing radiation (0, 40, 400, and 800 cGy) on the expression of transforming growth factor-beta1 (TGF-beta1), vascular endothelial growth factor (VEGF), and alkaline phosphatase. In addition, the authors studied the effects of ionizing radiation on MC3T3-E1 cellular proliferation and the ability of conditioned media obtained from control and irradiated cells to regulate the proliferation of bovine aortic endothelial cells. Finally, the authors evaluated the effects of adenovirus-mediated TGF-beta1 gene therapy in an effort to "rescue" irradiated osteoblasts. The exposure of osteoblast-like cells to ionizing radiation resulted in dose-dependent decreases in cellular proliferation and promoted cellular differentiation (i.e., increased alkaline phosphatase production). Additionally, ionizing radiation caused dose-dependent decreases in total TGF-beta1 and VEGF protein production. Decreases in total TGF-beta1 production were due to a decrease in TGF-beta1 production per cell. In contrast, decreased total VEGF production was secondary to decreases in cellular proliferation, because the cellular production of VEGF by irradiated osteoblasts was moderately increased when VEGF production was corrected for cell number. Additionally, in contrast to control cells (i.e., nonirradiated), conditioned media obtained from irradiated osteoblasts failed to stimulate the proliferation of bovine aortic endothelial cells. Finally, transfection of control and irradiated cells with a replication deficient TGF-beta1 adenovirus before irradiation resulted in an increase in cellular production of TGF-beta1 protein and VEGF. Interestingly, this intervention did not alter the effects of irradiation on cellular proliferation, which implies that alterations in TGF-beta1 expression do not underlie the deficiencies noted in cellular proliferation. The authors hypothesize that ionizing radiation-induced alterations in the cytokine profiles and differentiation states of osteoblasts may provide insights into the cellular mechanisms underlying osteoradionecrosis and impaired fracture healing. PMID- 11039377 TI - Preservation of a digital osteotendinous structure with an omental flap. AB - Taking into account the angiogenic properties of the omentum to revascularize ischemic tissues, this experimental, longitudinal, prospective, double-blind study in rabbits was designed to revascularize and preserve the mobility of a digital osteotendinous structure surgically devascularized in advance and to compare such omental angiogenic ability with that of the muscle and the panniculus carnosus. Thirty New Zealand rabbits were used. Three toes from the hind feet were surgically amputated from each rabbit. The skin was removed, exposing the bones, tendons, ligaments, and joints, to form what we termed the osteotendinous structure. Through a median laparotomy, the first part of each rabbit's own osteotendinous structure was placed inside the panniculus carnosus (group I), the second under the rectus abdominis muscle (group II), and the third was wrapped in a pediculate omental flap (group III). Three weeks later, each structure was assessed clinically for mobility and fibrosis and microscopically for fibrosis, newly formed vessels, viability, and tissue regeneration. Clinically, the group I structures showed a greater amount of fibrosis. The structures in groups II and III showed minimal fibrosis in all but four cases, which showed moderate fibrosis. Differences in joint mobility were assessed with the Kruskal-Wallis test. There was a statistically significant difference in mobility for the structures from group III, which was higher, followed by those from groups II and I. The exception was the proximal interphalangeal joints in groups II and III, for which the differences had no statistical significance. Microscopically, fibrosis and tissue necrosis were intense in the structures in group I, moderate in the group II structures, and mild in the group III structures. Conversely, vessel neoformation and tissue regeneration were intense in the structures in group III, moderate in group II, and were nil in group I. This study confirms with statistical significance that, in the rabbit, the omentum has a higher ability to revascularize degloved tissues than do the muscle and the panniculus carnosus, thus preserving a higher joint and tendon mobility. Consequently, it is suggested that a free omental flap be used in the treatment of ring avulsion injuries that lead to degloving of the digits. PMID- 11039378 TI - Acute squamous cell carcinoma arising within a recent burn scar in a 14-year-old boy. AB - We have described the case of a 14-year-old boy who developed an acute squamous cell carcinoma within a healed burn scar 6 weeks after thermal injury. This is a rare condition necessitating early excision and histopathologic confirmation of clearance. The authors recommend early skin grafting of deep partial- and full thickness burns to prevent future malignant transformation and a low index of suspicion of any nodules or ulceration appearing within a healed burn. PMID- 11039379 TI - "Ring" lipoma causing extensor tenosynovitis. PMID- 11039380 TI - Treatment of sternal nonunion with the Dall-Miles cable system. AB - We have used this technique in two patients. One had early sternal dehiscence with presternal infection, and the other had late sternal nonunion. Uncomplicated sternal union was achieved in both patients. The cables were nonpalpable in both patients, but they were removed in one patient at that patient's request. This method of using Dall-Miles cerclage cables is a straightforward and efficacious method of open reduction and internal fixation of the sternum. It is indicated for patients with chronic sternal nonunion or early postoperative separation of the sternal fragments and may be used even in the presence of an infection limited to the presternal space after adequate debridement and irrigation have been performed. Any recurrent superficial infection, although unlikely, can be cured by hardware removal after osseous union has been obtained. For sternal separation without fractures, four cables may simply be placed around the sternal halves and their tension increased. In the case of sternal fractures, the cables may be placed in figures of eight or in other woven configurations as needed for each individual case. PMID- 11039381 TI - The combined use of flaps based on subscapular vascular system for unilateral facial deformities. PMID- 11039382 TI - Clinical anthropometry and canons of the face in historical perspective. AB - Measurements of the human face as part of the body have been performed since the Greek era, and many aspects of ancient measurements can be found in modern clinical anthropometry. A historical appraisal of the use of facial measurements is presented. The influence on modern facial anthropometry of Greek proportion sciences, the golden proportion, canons of important Renaissance artists, physical anthropology, and cephalometry are discussed. The main difference between human measurements in classic times and modern anthropometry is the denial of realistic sizes and proportions in former times. Human forms and canons were depicted in a way the artist or scientist preferred, rather than how they objectively were. For reconstructive and cosmetic surgery, realistic sizes and proportions are assessed using anthropometric techniques and used as guidelines to correct deformities or disproportions. PMID- 11039383 TI - The surgical treatment of brachial plexus injuries in adults. AB - Posttraumatic brachial plexus palsy is a severe injury primarily affecting young individuals at the prime of their life. The devastating neurological dysfunction inflicted in those patients is usually lifelong and creates significant socioeconomic issues. During the past 30 years, the surgical repair of these injuries has become increasingly feasible. At many centers around the world, leading surgeons have introduced new microsurgical techniques and reported a variety of different philosophies for the reconstruction of the plexus. Microneurolysis, nerve grafting, recruitment of intraplexus and extraplexus donors, and local and free-muscle transfers are used to achieve optimal outcomes. However, there is yet no consensus on the priorities and final goals of reconstruction among the various centers. PMID- 11039384 TI - Training the next generation of plastic surgeons: a lesson from geese. PMID- 11039385 TI - Laser blepharoplasty with transconjunctival orbicularis muscle/septum tightening and periocular skin resurfacing: a safe and advantageous technique. AB - Carbon dioxide (CO2) laser blepharoplasty with orbicularis oculi muscle tightening and periorbital skin resurfacing is a safe procedure that produces excellent aesthetic results and diminishes the occurrence of complications associated with skin and muscle resection in the lower lid, particularly permanent scleral show and ectropion. The authors present a review of 196 cases of carbon dioxide laser blepharoplasty and periocular laser skin resurfacing performed at their center from April of 1994 to September of 1998. Of these cases, 113 patients underwent four-lid blepharoplasty, 59 underwent upper lid blepharoplasty only, and 24 underwent lower lid blepharoplasty only. Prophylactic lateral canthopexy was performed in 24 patients. Concomitant procedures (brow lift/rhytidectomy/rhinoplasty) were performed in 92 patients. The carbon dioxide laser blepharoplasty procedure resulted in no injuries to the globe, cornea, or eyelashes. Combined with laser tightening of the orbicularis oculi muscle and septum and periocular skin resurfacing, the transconjunctival approach to lower blepharoplasty preserves lower lid skin and muscle. Elimination of the traditional scalpel skin/muscle flap procedure results in a dramatically lower complication rate, particularly with regard to permanent ectropion and scleral show. Laser shrinkage of the orbicularis muscle and septum through the transconjunctival incision enables the correction of muscle aging changes such as orbicularis hypertrophy and malar festoons. The addition of periocular resurfacing enables the correction of skin aging changes of the eyelid that are not addressed by traditional scalpel blepharoplasty. In addition, lateral canthopexy constitutes an important adjunct to the laser blepharoplasty procedure for the correction of lower lid canthal laxity. PMID- 11039386 TI - Anterior cervicoplasty in the male patient. AB - Most men develop visible redundant tissue in the anterior neck with aging. Some seek surgical improvement. If the patient does not wish to have a conventional face/neck lift, anterior cervicoplasty is a good option. The procedure accomplishes tightening in the horizontal direction by excising a midline vertical ellipse of skin and subcutaneous fat. The surgeon tightens and lengthens the platysma muscles by suturing the anterior borders of the muscle to each other and by performing one or more Z-plasties in the muscle. A Z-plasty in the skin and subcutaneous tissue predictably creates a mental/cervical crease or angle with precise planning of the location of the horizontal limb. It also provides added length to the vertical skill closure. Every patient has thought that the improved contour of his neck more than offset the presence of a visible scar. In fact, no patient has indicated that his scar has been noticed by others, nor has any patient requested scar revision. PMID- 11039387 TI - Temporalis muscle hypertrophy: a new plastic surgery procedure. AB - Bilateral hypertrophy of the temporal muscle can give the impression of a harsh facial appearance that manifests itself as a morphopsychological conflict for the subject involved (Minotaur syndrome). This article describes a new facial aesthetic surgical procedure in the area of the temporal muscle. The author describes the surgical technique and the surgical instrument that he developed specifically for performing aesthetic contouring of the temporal area by reducing the muscle volume discrepancy ("myosuction"). The follow-up results of 11 cases demonstrated that this procedure renders valid results in the overall aesthetic reharmonization of the face and an improvement of individual psychological well being. PMID- 11039388 TI - The course of the inferior alveolar nerve in the normal human mandibular ramus and in patients presenting for cosmetic reduction of the mandibular angles. AB - This study was undertaken to quantify the path of the inferior alveolar nerve in the normal human mandible and in the mandibles of patients presenting for cosmetic reduction of the mandibular angles. The goals were: (1) to provide normative information that would assist the surgeon in avoiding injury to the nerve during surgery; (2) to characterize gender differences in the normal population; and (3) to compare the course of the nerve in the normal population to its course in a group of patients who presented with a complaint of "square face." The study was based upon the computerized tomographic scans of 10 normal patients (six men, four women) and 8 patients (all women) complaining of "square face." Using AnalyzePC 2.5 imaging software, the mandibles were segmented and the position of the nerve was recorded within its osseous canal in the mandibular ramus on each axial slice in which it was identifiable. Distances were calculated between the nerve and the anterior, posterior, lateral, and medial cortices. The positions of the lateral ramus prominence and the lowest point on the sigmoid notch were also recorded. The position of the mental foramen was recorded in relation to the nearest tooth, and the three-dimensional surface distances from the foramen to the alveolar bone, the inferior border of the mandible, and the mandibular symphysis were determined. The distances from the entrance of the nerve into the mandible to the lateral ramus prominence and the lowest point on the sigmoid notch were calculated. Summary statistics were obtained, comparing differences in gender. The nerve was identifiable in each ramus over a mean distance of 12.7 mm. On average, the lateral ramus prominence was 0.3 mm higher on the caudad-cephalad axis than the point at which the nerve entered the bone, whereas the location of the lowest point on the sigmoid notch was 16.6 mm above the nerve. The average distances from the nerve to the anterior, posterior, medial, and lateral cortices were 11.6, 12.1, 1.8, and 4.7 mm, respectively. Gender differences were significant for all of these except the medial cortex to nerve distance. On average, the mental foramen exited the body of the mandible immediately below the second premolar and the average surface distances from the foramen to the symphysis, the most cephalad alveolar bone, and the inferior border of the body were 30.9, 14.2, and 19.3 mm, respectively. With regard to the patients presenting for mandibular angle reduction, there were a few statistically significant but small scalar differences from normal controls. PMID- 11039389 TI - Transumbilical endoscopic breast augmentation: submammary and subpectoral. AB - Endoscopic techniques have recently been applied to aesthetic cosmetic surgery procedures. Endoscopic bilateral augmentation mammaplasty through a transumbilical approach ("TUBA") has recently been advocated as an alternative technique. The purpose of this article is to describe the author's transumbilical technique, to identify procedural limitations and special considerations, and to retrospectively analyze preliminary results. Five hundred thirteen patients (n = 1026 breasts) who underwent submammary transumbilical augmentation from January of 1993 through December of 1998 were evaluated. In 1997, the technique was further developed to permit subpectoral placement of implants; an additional 140 patients (n = 280 breasts) who underwent subpectoral transumbilical augmentation from September of 1997 through February of 1999 will also be presented. Success of the technique was based upon a number of criteria, including completion of the operation without conversion to an inframammary incision or reoperation, normal nipple-areola sensation, absence of hematoma formation, absence of infection, no umbilical scar revision, and patient satisfaction. Complications included hematoma (n = 2 breasts), conversion to inframammary incision (n = 5 breasts), and required secondary corrective procedure (n = 3 breasts). The majority of these complications occurred early in the learning curve. The successful augmentation rate in 1306 breasts was 99.2 percent. Based upon these results, transumbilical endoscopic breast augmentation is believed to be a safe alternative technique with excellent results. PMID- 11039390 TI - National plastic surgery survey: face lift techniques and complications. AB - The purpose of this study was to assess trends in technique and philosophy of face lifting, associated procedures, and the incidence and management of complications. Surveys were sent to 3800 members of the American Society of Plastic and Reconstructive Surgeons (ASPRS); 570 surveys (15 percent) were returned. Numerous very specific technique and philosophy questions were asked. Details of demographics, techniques, incidence of complications, management of complications, and basic philosophy are presented. Three basic conclusions can be gleaned from this study: (1) Surgeons perform more tried and true methods of aesthetic surgery, rather than the many new methods that seem to get the most attention in the media and at the meetings. (2) It seems that less-experienced surgeons tend to be generally more conservative in their approach to aesthetic surgery. (3) Complication rates reported by the plastic surgery community at large coincide with previous complication rates, as outlined in other nonsurvey studies. The authors expect to report additional data from the survey--on brow surgery (part II) and facility and ancillary procedures (part III)--in forthcoming publications. PMID- 11039391 TI - Liposuction as an adjunct to a full abdominoplasty revisited. PMID- 11039392 TI - A 1927 view of cosmetic surgery. PMID- 11039393 TI - Deaths associated with liposuction. PMID- 11039394 TI - Frontalis musculotaneous island flap for coverage of forehead defect. PMID- 11039395 TI - Eyelid crutches for ptosis: a forgotten solution. PMID- 11039396 TI - Endoscopic carpal tunnel release instruments used for auricular cartilage scoring and correcting a flattened antihelix. PMID- 11039397 TI - Long-term complications of facial injections with Restylane (injectable hyaluronic acid). PMID- 11039398 TI - Latissimus dorsi myocutaneous flap reconstruction of neck and axillary burn contractures. PMID- 11039399 TI - Fat necrosis after a transverse rectus abdominis myocutaneous flap? How about massage? PMID- 11039400 TI - Avoiding the teardrop-shaped nipple-areola complex in vertical mammaplasty. PMID- 11039401 TI - Zoster following immediate transverse rectus abdominis myocutaneous breast reconstruction. PMID- 11039402 TI - Coincidental breast carcinoma detection in reduction mammaplasty specimens. PMID- 11039403 TI - Protecting hand splints. PMID- 11039404 TI - How to deal with the umbilical stalk during abdominoplasty. PMID- 11039405 TI - Umbilical transposition: an unnecessary step. PMID- 11039406 TI - Conservative treatment of early-stage penile melanoma. PMID- 11039407 TI - Use of the stoma bag in a patient with buttock burn. PMID- 11039408 TI - Combined use of V-Y advancement flap and rotation flap. PMID- 11039409 TI - Treating seromas. PMID- 11039410 TI - Repair of tips of microforceps. PMID- 11039411 TI - Simple suction device for autologous epidermal grafting. PMID- 11039412 TI - Tissue adhesives: more than just a sticky situation. PMID- 11039413 TI - A wound after contact between a child and a dog does not necessarily mean an animal bite. PMID- 11039414 TI - Effects of nerve growth factor on nerve regeneration through a vein graft across a gap. PMID- 11039415 TI - Results of macular hole surgery in patients over 80 years of age. AB - PURPOSE: To evaluate the results of macular hole surgery in patients over 80 years of age to determine if surgery is beneficial in an elderly patient population. METHODS: Twenty consecutive patients over age 80 with idiopathic macular holes and 20 nonconsecutive controls younger than 80 years treated for idiopathic macular holes were compared in a retrospective, consecutive case control series. RESULTS: The macular hole was closed 3 months after surgery in 19/20 eyes (95%) of patients over age 80 years and 17/20 eyes (85%) of patients under 80 years. The mean preoperative visual acuity was 20/160 in patients over 80 years and 20/160-1 in patients under 80 years. The mean visual acuity at 3 months was 20/63 in eyes of patients over 80 years and 20/80 in eyes of patients under 80 years (P = 0.3). The mean visual acuity was 20/50-2 in eyes of patients over 80 years and 20/63-2 in eyes of patients under 80 years at the final examination (P = 0.403). CONCLUSIONS: Macular hole surgery is beneficial in patients over 80 years of age, with very similar results to those of patients younger than 80 years of age. Age should not be the primary criterion for recommending macular hole surgery. PMID- 11039416 TI - Endolaser around macular hole in the management of associated retinal detachment in highly myopic eyes. AB - PURPOSE: To analyze the role of endolaser around macular hole in managing associated retinal detachment in patients with high myopia. METHODS: Review of medical records of 25 consecutive eyes of 25 patients with at least 5.00 diopters of myopia who underwent primary pars plana vitrectomy and fluid-gas exchange. In the first half of the study period, one row of contiguous argon green endolaser was routinely applied over the retinal edge of the macular hole (EL group). In the second half of the study period, endolaser was not applied around any macular hole (NEL group). Demographic information, intraoperative and postoperative complications, and final visual acuities and retinal reattachment rates were studied. RESULTS: The mean (+/-SD) refractive error was -11.8 +/- -3.5 diopters (D) for the EL group and -11.6 +/- -5.4 D for the NEL group. The mean axial length was 29.0 +/- 1.8 mm for the EL group and 28.3 +/- 1.7 mm for the NEL group. The primary anatomic success was 62.5% (10/16) and 77.8% (7/9) in the EL and NEL groups, respectively. No statistically significant difference was found in preoperative, postoperative, or change in best-corrected visual acuities between the two groups. CONCLUSION: Endolaser around the macular hole after pars plana vitrectomy and internal gas tamponade may not affect the anatomic or visual outcome in primary retinal detachment secondary to a highly myopic macular hole. PMID- 11039417 TI - Macular buckling for retinal detachment due to macular hole in highly myopic eyes with posterior staphyloma. AB - PURPOSE: To evaluate the efficacy of macular buckling surgery using a special exoplant. METHODS: The authors reviewed 33 cases of patients with retinal detachment due to macular hole with posterior staphyloma who underwent a macular bucking procedure and 11 cases of patients with the same condition due to macular hole without posterior staphyloma or due to a hole along the edge of the posterior staphyloma who underwent gas injection or vitrectomy. The reattachment rate, visual acuity (VA), and area size (V-4, I-4 isopter) by Goldmann perimetry were calculated. RESULTS: Reattachment rate for the macular buckling procedure was 94% (initial) and 100% (final), and that for the gas injection or vitrectomy was 100% (both initial and final). Twenty of the 33 eyes (60.6%) had VA of 20/400 or better, and none of these 33 eyes had a VA at the level of finger counting or poorer following macular buckling. Nine of the 11 eyes (81.8%) had VA of 20/400 or better, and none of these 11 eyes had a VA at the level of finger counting or poorer following the gas injection or vitrectomy. The area size of V-4 isopter in the macular buckling group was significantly larger than that in the gas injection or vitrectomy group (P = 0.017). CONCLUSION: The macular buckling procedure is effective when considered both from an anatomic and a functional perspective for retinal detachment due to macular hole with posterior staphyloma. PMID- 11039418 TI - Optical cross-sectional evaluation of successfully repaired idiopathic macular holes by retinal thickness analyzer. AB - PURPOSE: To describe optical cross-sectional images of successfully repaired idiopathic macular holes and to determine the influences of epiretinal membranes and retinal pigment epithelial (RPE) changes on foveal reconstruction and the correlation between retinal thickness and postoperative visual acuity. METHODS: In a prospective study, optical cross-sectional imaging and retinal thickness measurement of the macula using a retinal thickness analyzer were performed on 63 eyes of 63 patients who underwent successful macular hole surgery. RESULTS: Cross sectional images of foveal reconstruction were morphologically categorized into four patterns: normal fovea (23 eyes [37%]), cavernous fovea (19 eyes [30%]), flat fovea (11 eyes [17%]), and irregular fovea (10 eyes [16%]). Epiretinal membranes were observed in the last two groups (55% and 40%) and RPE changes were observed only in the irregular fovea group (16%). The mean retinal thickness of the fovea in all eyes was 213 +/- 92 microm (mean +/- SD; range, 93-570 microm), which varied significantly (P < 0.001) among the four groups. Linear regression analysis showed a significant correlation between retinal thickness at the fovea and logarithmic converted visual acuity (R2 = 0.42, P < 0.001). CONCLUSIONS: Structural features of foveal reconstruction following successful macular hole surgery involved four patters: normal fovea, cavernous fovea, flat fovea, and irregular fovea. Retinal thickness of the fovea, which varied among the groups, correlated with postoperative visual acuity. Postoperative epiretinal membrane formation and RPE damage may disturb normal foveal reconstruction and visual recovery. PMID- 11039419 TI - Primary vitrectomy alone for repair of retinal detachments following cataract surgery. AB - PURPOSE: To report the visual, anatomic, and refractive results of primary vitrectomy alone for the repair of retinal detachments (RD) following cataract surgery. METHODS: Retrospective review of office charts and operative reports of 83 eyes. RESULTS: A minimum of 4 months' follow-up was achieved for 78 pseudophakic or aphakic eyes that underwent primary vitrectomy, internal drainage of subretinal fluid, retinopexy, and intravitreal gas injection for RD repair. Anatomic reattachment was achieved in 93.6% of cases after one procedure and in 96.2% eventually. Median preoperative Snellen acuity was 20/200 and increased to 20/25 at final examination. For the 45 eyes with macula-off detachments, 80% achieved final acuities greater than or equal to 20/40. The average refractive change following surgery was -0.11 diopters. Transient postoperative ocular hypertension was seen in 17.9% and proliferative vitreoretinopathy with recurrent RD in 5.1%. CONCLUSION: Primary pars plana vitrectomy is a highly effective treatment modality for the repair of RD following cataract surgery, and appears to be refractively neutral. PMID- 11039420 TI - Factors related to subretinal proliferation in patients with primary rhegmatogenous retinal detachment. AB - PURPOSE: To evaluate factors related to subretinal proliferation in patients with primary rhegmatogenous retinal detachment (RD). METHODS: A total of 262 consecutive patients (267 eyes) with rhegmatogenous RD were evaluated retrospectively to determine factors associated with subretinal proliferation. RESULTS: Logistic regression analysis was used to identify the following factors associated with subretinal proliferation that caused retinal traction: number of quadrants detached (adjusted odds ratio [AOR] = 2.21), longer duration of RD (AOR = 1.52), younger patient age (AOR = 0.51), and presence of atrophic retinal breaks (AOR = 0.41). CONCLUSION: Atrophic retinal breaks, younger patient age, and RD of long duration and greater extent were associated with the presence of subretinal proliferation. PMID- 11039421 TI - Bioadhesives for intraocular use. AB - BACKGROUND/PURPOSE: A safe, effective adhesive could be useful in the management of retinal holes or tears and selected complicated retinal detachments, as well as for attaching a small electronic device (retinal prosthesis) to the retina. In this study, we examined nine commercially available compounds for their suitability as intraocular adhesives. METHODS: The following materials were studied: commercial fibrin sealant, autologous fibrin, Cell-Tak, three photocurable glues, and three different polyethylene glycol hydrogels. An electronic strain gauge measured the adherence forces between different glues and the retina. The stability of hydrogels at body temperature and the impermeability of the hydrogel adhesive to dextran blue were examined. Long-term biocompatibility testing of the most promising glues in terms of adhesive force, consistency, and short-term safety (hydrogels) were done in rabbits. Funduscopy, electroretinogram, and histology of the retina were performed. RESULTS: Hydrogels had 2 to 39 times more adhesive force (measured in mN) than the other glues tested. They liquefied at body temperature after 3 days to a few months. Hydrogels were impermeable to dextran blue. One type of hydrogel proved to be nontoxic to the retina. CONCLUSIONS: Hydrogels proved to be superior for intraocular use in terms of consistency, adhesiveness, stability, impermeability, and safety. PMID- 11039422 TI - Anesthetic myotoxicity as a cause of restrictive strabismus after scleral buckling surgery. AB - PURPOSE: To explore the possibility that anesthetic myotoxicity may play a role in restrictive strabismus following scleral buckling procedures. METHODS: The authors performed a retrospective study of patients who presented with strabismus following scleral buckling procedures. Details were sought regarding the scleral buckling procedure, including type and route of anesthesia. The types of strabismus were compiled, as were relevant findings at strabismus surgery. The contributing vitreoretinal surgeons were surveyed regarding the usual type and route of anesthesia used for their scleral buckling procedures. RESULTS: Over 90% of scleral buckling procedures resulting in significant strabismus were performed under local anesthesia. Of the 17 patients on whom strabismus surgery was performed, 14 had positive forced ductions. A hypodeviation of the buckled eye was the most common presentation. CONCLUSION: Based on the types, patterns, and amounts of strabismus encountered after scleral buckling procedures, and the similarity of these findings to cases of strabismus following retrobulbar anesthesia for cataract procedures, the authors propose that local anesthetic myotoxicity is often the primary cause of strabismus occurring after scleral buckling procedures for retinal detachment. PMID- 11039423 TI - Influence of membrane differential filtration on the natural course of age related macular degeneration: a randomized trial. AB - PURPOSE: Membrane differential filtration is able to optimize rheologic parameters by eliminating high molecular weight proteins and lipoproteins from the blood. Following the hypothesis that these changes result in an improvement of the microcirculation, the authors tested the efficacy of membrane differential filtration in improving visual function in patients with age-related macular degeneration (ARMD). METHODS: Forty patients (40 eyes) were randomized into two groups. The treatment group was treated five times over a period of 21 weeks. In both groups, 9/20 of the eyes showed subfoveolar subretinal neovascularization. The main parameter of the study was visual acuity (VA). Electroretinogram (ERG), electrooculogram, and macular visual evoked potentials were also recorded. Plasma and whole blood viscosity and erythrocyte aggregation were measured. RESULTS: The 20 patients treated repeatedly over a period of 21 weeks showed a mean improvement of 0.63 lines (SD 1.8) of VA on Early Treatment Diabetic Retinopathy Study charts. The control group showed a deterioration of 0.94 lines (SD 1.7) compared to VA at baseline examination. The amplitude of the ERG photopic a-wave and the flicker ERG was significantly increased. The rheologic parameters were lowered in all treated patients. CONCLUSION: Repetitive treatment with membrane differential filtration is able to improve visual acuity of patients with ARMD and the natural course of this disease. Several questions arise from the results of this study. Further research will show if it is possible to optimize the selection of patients for subgroups with predictive responses through morphologic and functional tests and how to create an optimized and individual treatment strategy determined by the quality, intensity, and frequency of treatment sessions. PMID- 11039424 TI - Clinicopathologic correlation of fluorescein and indocyanine green angiography in exudative age-related macular degeneration. AB - PURPOSE: To correlate the clinical and histopathologic features of an eye with age-related macular degeneration studied with fluorescein (FA) and indocyanine green (ICG) angiography 4.5 months before the patient's death. METHODS: Histopathologic features from serial sections through the macula of a 76-year-old man with occult choroidal neovascularization (CNV) were reconstructed in a scaled two-dimensional map and compared with FA and ICG angiogram images obtained 4.5 months before his death. RESULTS: The region of prior laser photocoagulation was identified as a well-demarcated hypofluorescent region in the early frames of the FA and the early and late phases of the ICG angiogram. This corresponded histopathologically to a well-circumscribed area of absence of the choriocapillaris, loss of the outer retina and retinal pigment epithelium, and scarring of the choroid. Occult CNV characterized by elevated late hyperfluorescence on the FA and intense well-defined hyperfluorescence on the ICG angiogram corresponded to a thick fibrovascular membrane in the subretinal space and within Bruch's membrane. Thin extensions of both the subretinal and intra Bruch's membrane fibrovascular membrane components corresponded to nonelevated stippled late hyperfluorescence on the FA and mild late hyperfluorescence on the ICG angiogram. CONCLUSION: Histopathologic mapping revealed a large fibrovascular complex located subretinally and within Bruch's membrane with thin and thick components that correlate well with findings of occult CNV on FA and ICG angiography. PMID- 11039425 TI - Quantitative effect of intravitreally injected tissue plasminogen activator and gas on subretinal hemorrhage. AB - PURPOSE: To quantify the effect of intravitreally injected tissue plasminogen activator (TPA) and an expanding gas on freshly formed subretinal hemorrhage (SRH). PATIENTS AND METHODS: Thirteen patients with acute (1 week or less) SRH due to age-related macular degeneration (ARMD) were treated with an intravitreal injection of 50 microg TPA, and 24 hours later, with an expanding gas. Fundus photographs taken before and 24 hours after each injection were digitized and calibrated. Area, geometric center, and shift of the SRH were measured at each time point using NIH image analysis software. Elevation of the SRH was assessed by echography. RESULTS: Compared to the preoperative size, SRH enlarged significantly 24 hours after TPA injection (P < 0.001). A significant shift of the geometric center toward the inferior retinal periphery out of the macula was found after gas injection (P < 0.001). Significant horizontal displacement of the SRH was not noted after TPA or gas injection. More elevated subretinal blood clots showed a larger increase in size after TPA injection than flat clots (P < 0.05). CONCLUSION: Enlargement of SRH in a gravity-dependent manner indicates subretinal liquefaction of the clot after TPA treatment. An intravitreal injection of gas 24 hours later can significantly displace the SRH inferiorly. PMID- 11039426 TI - Histopathologic abnormalities in rabbit retina after intravitreous injection of expansive gases and air. AB - PURPOSE: To evaluate histopathologic retinal changes in rabbit eyes after injection of pure perfluoropropane (C3F8) gas, pure sulfur hexafluoride (SF6) gas, or air into the vitreous cavity. METHODS: Air, C3F8 gas, or SF6 gas (0.4 mL each) were injected into rabbit vitreous cavities. Two, 4, and 6 weeks later, light and electron microscopic examinations were conducted, and the immunohistochemical localization of glutamate in the retina was studied. Noninjected eyes served as controls. RESULTS: At all time points, thinning or disappearance of the outer plexiform layer in the superior retina in eyes that received C3F8 gas was found; the inferior retina was the same as in controls. In eyes that received SF6 gas or air, light and electron microscopy showed that the superior and inferior retina were the same as in controls at all time points. Immunohistochemical examination showed abnormal glutamate distribution of the superior retina in eyes injected with C3F8 gas, SF6 gas, or air. However, glutamate distribution was the same as in controls in the inferior retina in eyes injected with C3F8 gas, SF6 gas, or air. CONCLUSIONS: Retinal tamponade using intraocular gases induces histopathologic retinal changes in the superior retina of the rabbit eye, where the gases are in continuous contact with the eye. PMID- 11039427 TI - Pharmacokinetics of the fluocinolone/5-fluorouracil codrug in the gas-filled eye. AB - PURPOSE: To determine the effect of intraocular gas tamponade on drug levels achieved with the intravitreal sustained-release fluocinolone (FL)/5-fluorouracil (5-FU) codrug pellet. METHODS: After insertion of a 10-mg codrug pellet into the right eyes of 43 New Zealand white rabbits, perfluoropropane (0.4 mL of 100% C3F8) or a control sham was then injected into the midvitreous cavity. On postoperative days 2, 4, 7, 21, and 42, aqueous samples were collected, the rabbits were killed, and the right eyes were enucleated. The vitreous and remaining codrug pellet were then isolated. Pellet and intravitreal drug levels were determined by high-pressure liquid chromatography. RESULTS: No measurable drug levels were detected in any of the aqueous samples. Maximal gas expansion occurred by day 4 and partial resorption was observed by days 14 to 21. Vitreous FL and 5-FU levels during C3F8 expansion (day 2) were statistically significantly higher in the gas-filled eyes. On postoperative days 4, 7, 21, and 42, there were no statistically significant differences between FL and 5-FU drug levels in eyes containing C3F8 as compared with control eyes. Pellet codrug, FL, and 5-FU levels over time were similar in gas-filled and control eyes. CONCLUSIONS: Intraocular gas tamponade does not significantly affect the sustained intravitreal drug levels achieved with the FL/5-FU codrug. If clinically efficacious, the FL/5-FU codrug formulation does not need to be altered to treat proliferative vitreoretinopathy in the presence of intraocular gas. PMID- 11039428 TI - Efficacy of the ganciclovir implant in the setting of silicone oil vitreous substitute. AB - OBJECTIVE: To evaluate the efficacy of the ganciclovir implant in the setting of silicone oil vitreous substitute. MATERIALS AND METHODS: The authors retrospectively reviewed the charts of 19 patients with cytomegalovirus retinitis who had both a ganciclovir implant and silicone oil vitreous substitute. None of the patients was receiving highly active antiretroviral therapy during the study period. Kaplan-Meier analysis was used to evaluate time to progression in eyes with the ganciclovir implant and silicone oil. RESULTS: In all eyes, the ganciclovir device implantation preceded or coincided with silicone injection. Kaplan-Meier analysis revealed that time to 25% failure from the date of the presence of both the implant and oil was 129 days. Time to 25% failure from the date of ganciclovir device implantation was 179 days. CONCLUSIONS: These results compare favorably with conventional treatment. The ganciclovir implant can be a useful treatment modality in eyes with silicone oil. PMID- 11039429 TI - Ultrasound biomicroscopy in silicone oil-filled eyes. AB - PURPOSE: To establish reproducible ultrasound biomicroscopy (UBM) imaging patterns characteristic of the presence and behavior of silicone oil and of peripheral vitreoretinal proliferation in eyes that have undergone vitreoretinal surgery with silicone oil injection. METHODS: The study design was a case series. Ultrasound biomicroscopy was performed on 34 eyes of 34 patients (age range, 20 68 years). For an image to be considered a UBM pattern, it was required to be consistently reproducible in the same eye and to be observed in more than one eye. RESULTS: Four patterns are described: silicone oil particles, ghost images, "painting" or surface impregnation of iris and anterior chamber angle, and peripheral proliferative vitreoretinopathy. One type of ghost image, the secondary reflection of the cornea, paired with disappearance of the aqueous silicone oil interface corresponded to massive silicone oil displacement into the anterior chamber. Gross proliferation produced funnel and tentlike images, whereas minute, networklike images seemed to represent early changes. There was a close correspondence between these UBM patterns and previous ophthalmologic clinical findings. CONCLUSION: Easily recognizable, distinct UBM patterns are characteristic of conditions found in silicone oil-filled eyes. Ultrasound biomicroscopy seems to be a useful tool for assessing changes in silicone oil filled eyes. PMID- 11039430 TI - Histopathologic retinal changes with intravitreous fluorosilicone oil in rabbit eyes. AB - BACKGROUND: Fluorosilicone oil (FSiO, 1,000-10,000 centistokes), which has a higher density (1.29 g/cm3) than vitreous gel, is useful as an operative tool and a tamponade for the inferior retina during difficult retinal detachment surgery. The occurrence of histopathologic retinal changes after injection of FSiO into the vitreous cavity is controversial. METHODS: Retinas obtained from 18 rabbits were evaluated histopathologically within 8 weeks of injection into the vitreous cavity of purified FSiO or balanced salt solution as control in phakic eyes. The histopathologic retinal changes caused by FSiO were compared with those of previously reported high-density hydrophobic vitreous substitutes, such as silicone-fluorosilicone copolymer oil (SiFO) and perfluorocarbon liquids. RESULTS: By light and electron microscopy, all retinas injected with FSiO were the same as control retinas within 2 weeks of the injection, but the outer plexiform layer disappeared from the inferior retina 4 weeks after injection. The receptor cell nuclei migrated to the photoreceptor layer in the inferior retina 8 weeks after injection. However, no preretinal membrane, including foam cells, was found in any eye injected with FSiO. CONCLUSION: FSiO may be useful as a temporary vitreous substitute in difficult inferior retinal detachments. PMID- 11039431 TI - Diagnostic and therapeutic challenges. PMID- 11039432 TI - Diagnostic and therapeutic challenges. PMID- 11039433 TI - Herpes simplex virus type 2 acute retinal necrosis in a patient with systemic lupus erythematosus. PMID- 11039434 TI - Subfoveal choroidal neovascular membrane excision in Vogt-Koyanagi-Harada syndrome. PMID- 11039435 TI - Iridocorneal apposition after vitrectomy and gas injection. PMID- 11039436 TI - Plurifocal cilioretinal occlusion as the presenting symptom of cardiac myxoma. PMID- 11039437 TI - Presumed choroidal--retinal arteriole anastamosis after penetrating ocular trauma. PMID- 11039438 TI - Triamcinolone acetonide as an aid to visualization of the vitreous and the posterior hyaloid during pars plana vitrectomy. PMID- 11039439 TI - Retrobulbar chlorpromazine in blind and seeing painful eyes. PMID- 11039440 TI - Incontinentia pigmenti: a florid case with a fluminant clinical course in a newborn. PMID- 11039441 TI - Improvement in macular function after epiretinal membrane removal in a patient with Stargardt disease. PMID- 11039442 TI - Diffuse subretinal fibrosis syndrome. PMID- 11039443 TI - Topical anesthesia in posterior vitrectomy. PMID- 11039444 TI - Topical anesthesia in posterior vitrectomy. PMID- 11039445 TI - Topical anesthesia in posterior vitrectomy. PMID- 11039446 TI - Primary intraocular lens implantation during pars plana vitrectomy and intraretinal foreign body removal. PMID- 11039447 TI - An adjustable-curvature endolaser probe for vitreoretinal surgery. PMID- 11039448 TI - A new extendable flexible pick for subretinal dissection. PMID- 11039449 TI - Choroidal infarction after embolization of arteriovenous fistula of middle meningeal artery. PMID- 11039450 TI - A case of double idiopathic macular holes. PMID- 11039451 TI - The BNMS Millennium Highlights Lecture. British Nuclear Medicine Society. PMID- 11039452 TI - FDG PET in the evaluation of the aggressiveness of pulmonary adenocarcinoma: correlation with histopathological features. AB - 2-[Fluorine-18]fluoro-2-deoxy-d-glucose (FDG) uptake within the primary lesion correlates with survival on positron emission tomography (PET) studies of patients with non-small cell lung cancer. The more metabolically active the tumour, the worse the outcome. The aim of this study was to determine whether a correlation exists between aggressiveness as determined by pathology and the findings of FDG PET in pulmonary adenocarcinoma. Thirty-five patients with 38 adenocarcinomas of the lung were studied. All patients underwent thoracotomy within 4 weeks of the FDG PET study. For semiquantitative analysis, standardized uptake values (SUVs) were calculated. Patients were classified into high SUV (> or = 4.0) and low SUV (<4.0) groups. The degree of FDG uptake (SUVs) in primary lung lesions was correlated with the histopathological features of aggressiveness (pleural involvement, vascular invasion or lymphatic permeation). The mean SUV of aggressive adenocarcinomas (4.36+/-1.94, n = 22) was higher than that of non aggressive ones (1.53+/-0.88, n = 16) (P < 0.0001). Tumours with a high FDG uptake have a significantly higher likelihood of aggressiveness than those with a low FDG uptake (P = 0.0004). Analysis by the Kaplan-Meier methods revealed that the groups had different prognoses (log-rank test, P = 0.0099). The high SUV group had a significantly worse prognosis. In conclusion, a correlation was seen between aggressiveness as determined by pathology and glucose metabolism as measured by FDG PET in adenocarcinoma of the lung. FDG PET may be used as a non invasive diagnostic technique in measuring aggressiveness and prognosis in patients with pulmonary adenocarcinoma. PMID- 11039453 TI - 18FDG SPECT to assess myocardial viability: initial experience at a hospital remote from a cyclotron. AB - 18F-Fluorodeoxyglucose positron emission tomography (18FDG PET) is the recognized gold standard for the assessment of myocardial viability, but is not widely available in the UK. FDG imaging on a gamma camera with high-energy collimators (FDG SPECT) has been shown to have an accuracy comparable with that of FDG PET for the assessment of myocardial viability. This study was performed to assess the feasibility of introducing FDG SPECT for myocardial viability at a hospital a considerable distance away from a cyclotron (200 miles). Twenty-three patients, who were being actively considered for revascularization but had demonstrated fixed defects on stress/rest with nitrate tetrofosmin imaging, underwent FDG SPECT. Image quality was acceptable in all patients. Nine out of the 23 patients with defects classed as fixed on tetrofosmin imaging demonstrated viability on FDG SPECT. Six of these nine patients, reported to have some viable myocardium on FDG SPECT, underwent revascularization as a result. This study has demonstrated that FDG SPECT is feasible at a site some distance from a cyclotron. PMID- 11039454 TI - A comparison of Tl-201 stress-reinjection-prone SPECT and Tc-99m-sestamibi gated SPECT in the differentiation of inferior wall defects from artifacts. AB - The frequency of false positive results obtained from the inferior myocardial region using single photon emission computed tomography (SPECT) myocardial perfusion scintigraphy is significantly higher than that obtained from other regions. Several methods, such as prone-position imaging, have been proposed to overcome this diagnostic problem. The aim of the present study was to compare the results of Tc-99m-sestamibi gated SPECT and Tl-201 prone SPECT in the differentiation of inferior wall artifacts from true defects. For this purpose, 38 subjects, whose coronary anatomies were documented on angiography, underwent same-day stress-rest Tc-99m-sestamibi gated SPECT and Tl-201 stress-reinjection prone (whose standard supine images demonstrated fixed defects on the inferior wall) SPECT. Gated SPECT was performed by 8 frames per cycle acquisition over a 180 degree rotation on 30 projections. Four gated SPECT slices were obtained on mid-ventricular vertical long axis, horizontal long axis and apical and basal short axis planes, and displayed in cine-format. Both Tl-201 prone imaging and Tc 99m-sestamibi gated analysis increased the specificity of inferior wall disease detection remarkably from 54% to 85% and 46% to 82%, respectively (P<0.05). The difference between diagnostic accuracies was not significant (80% and 82%, respectively) (P > 0.05). The positive predictive values for true defects were 96% for Tl-201 prone imaging and 94% for Tc-99m-sestamibi gated imaging. Based on segmental analysis, the two modalities showed fair agreement (kappa = 0.44 for standard supine protocols, kappa = 0.46 for Tl-201 prone and Tc-99m-sestamibi gated SPECT). It can be concluded that Tc-99m-sestamibi gated SPECT, requiring only two-step acquisition, may potentially increase the test specificity for coronary artery disease (CAD) of the inferior wall as well as does Tl-201 stress reinjection-prone SPECT. By giving functional information, it seems the most practical method in daily use for supplying the most extensive information about patients with suspected or known CAD. PMID- 11039455 TI - Thallium-201 myocardial SPET in patients with collagen disease. AB - We analysed stress 201Tl myocardial single photon emission tomography (SPET) in collagen disease patients to evaluate abnormal uptake patterns and their clinical significance in the assessment of the cardiac status of these patients. The main purpose of the study was to evaluate the clinical significance of reverse redistribution. Twenty-two collagen disease patients (13 with progressive systemic sclerosis (PSS) and nine with systemic lupus erythematosus (SLE)) were examined by 201Tl myocardial SPET with exercise (n = 9) or dipyridamole stress (n = 13). For quantitative analysis, each 201Tl SPET polar map was divided into 17 segments, and the 201Tl uptake pattern of each segment was classified into four types. Eighteen (82%) of the patients showed abnormal findings on 201Tl SPET. Of the 374 segments analysed, 295 (79%) were classified as normal, 16 (4%) as reverse redistribution, 49 (13%) as reversible defect and 14 (4%) as fixed defect. Patients were divided into two groups: those with cardiac abnormalities on conventional testing (Group A, n = 10) and those without (Group B, n = 12). The incidences of fixed defect, reversible defect and reverse redistribution were significantly higher (P <0.01, P <0.0005, P <0.05, respectively) in Group A than in Group B. Nine (90%) of the patients in Group A and nine (75%) in Group B showed abnormal findings. No significant difference was found between the PSS and SLE patients in the incidence of the individual uptake patterns. Stress 201Tl myocardial SPET appears to be an effective method of evaluating a wide spectrum of myocardial involvement in collagen disease patients and in assessing their clinical cardiac status. Reverse redistribution is found to be a significant finding in collagen disease patients. PMID- 11039456 TI - Renal transplantation for amyloid end-stage renal failure-insights from serial serum amyloid P component scintigraphy. AB - Although end-stage renal failure (ESRF) is common in systemic amyloidosis, few such patients receive renal transplants. Serum amyloid P component (SAP) scintigraphy is a specific method for the imaging and quantification of amyloid deposits in vivo, which has not previously been used to evaluate the outcome of renal transplantation in patients with amyloidosis. Evidence of renal graft amyloid was sought by SAP scintigraphy in 15 patients with systemic amyloidosis who had undergone renal transplantation 42-216 months (median, 73 months) previously. Prospective serial scans were obtained annually in eight cases. Renal grafts studied shortly after transplantation gave blood-pool images. The grafts remained normal in all patients whose underlying amyloidogenic disorder had remitted, whereas there was abnormal uptake of labelled SAP, indicating graft amyloidosis, in four out of 10 patients whose amyloid fibril precursor protein supply had not diminished. Graft amyloidosis was corroborated by renal dysfunction in each case, and by histology in one patient. SAP scintigraphy enables renal transplant grafts to be monitored noninvasively for involvement by amyloid. The lack of graft amyloidosis in all patients in whom the amyloidogenic underlying disorder had remitted, and in more than half of those in whom it had not, supports the use of renal transplantation for ESRF in systemic amyloidosis. PMID- 11039457 TI - Influence of time interval and number of blood samples on the error in renal clearance determination using a mono-exponential model: a Monte Carlo simulation. AB - Mono-compartmental analysis based on 2- and 4-h blood samples (BS) of 51Cr-EDTA (EDTA, ethylenediaminetetraacetic acid) is commonly used for the calculation of the glomerular filtration rate (GFR). The purposes of this study were to estimate the magnitude of error in clearance induced by errors in the time of sampling and activity measurement; to estimate the impact of changing the interval between the BS; and to assess the influence of a higher number of BS in reducing the error. A model of mono-exponential curves based on a finite number of BS was created. Normally distributed random errors were introduced in the time of sampling and activity measurement. In a first step, three different time intervals were used; in a second step, seven different numbers of BS were used, all taken between 120 and 240 min. For each setting, the random errors were successively introduced 200 times and the coefficients of variation (CV) of the calculated clearances were determined. Variable errors in clearance were induced by errors in the time of sampling and activity measurement. In general, the observed errors were higher for high and low clearance, with lower errors for moderately reduced clearances. The errors in indicating the time of sampling played an important role for high clearance, whereas the errors in activity measurements led to important errors for low clearance. Prolonging the interval from 1 to 2 h resulted generally in an important decrease in error, except in the range 60-100 ml x min(-1). Prolonging the interval from 2 to 3 h resulted in only a small additional decrease in error, except for very low clearance. Errors in indicating the time of sampling and in activity measurements induce errors in clearance determination. These errors cannot be significantly reduced by simply increasing the number of BS or by prolonging the interval between the samples. It is probably better, in most cases, to keep using the 2-4-h method and to take extreme care when indicating the time of sampling and when measuring the activity, instead of increasing the number of samples or lengthening the procedure. PMID- 11039458 TI - Comparison of Tc-99m-labelled antileukocyte fragment Fab' and Tc-99m-HMPAO leukocyte scintigraphy in the diagnosis of bone and joint infections: a prospective study. AB - Between January and July 1998, we conducted a prospective study to compare Tc-99m labelled antigranulocyte monoclonal antibody fragment Fab' (LEUKOSCAN) scintigraphy versus Tc-99m-hexamethylpropyleneamine oxime (Tc-99m-HMPAO)-labelled leukocyte scintigraphy (HMPAO-LS) for the diagnosis of unselected patients with bone and joint infection. Twenty-three patients (16 men and 7 women; mean age, 67 years) with suspected bone infection were explored successively with bone scintigraphy, HMPAO-LS and LEUKOSCAN scintigraphy. Thirty-two foci were studied (diabetic foot = 11, prosthetic material = 8, joint disease = 4, others = diagnosed in 18 cases, eight on the basis of bacteriological and histological examination of surgical or puncture specimens, with or without radiographic signs, and 10 on the basis of clinical course and radiographic findings. Overall sensitivity, specificity and accuracy were 86%, 72% and 78%, respectively, for LEUKOSCAN scintigraphy (12 true positives (TP), 13 true negatives (TN), 5 false positives (FP), 2 false negatives (FN)), 93%, 100% and 96%, respectively, for HMPAO-LS (13TP, 18TN, 0FP, 1FN), and 100%, 17% and 53.3%, respectively, for bone scintigraphy. In this small series, LEUKOSCAN scintigraphy was found to be less specific for the diagnosis of osteomyelitis than HMPAO-LS. In addition, the interpretation of LEUKOSCAN scintigraphy is more difficult than HMPAO-LS for the diagnosis of bone infection in the diabetic foot, and would appear to be less discriminating for differentiating soft tissue infection from osteitis in the case of plantar perforating ulcers. PMID- 11039459 TI - Enhancement of 131I-MIBG uptake in carcinoid tumours by administration of unlabelled MIBG. AB - Iodine-131 metaiodobenzylguanidine (131I-MIBG) has been used with success for the palliation of symptomatic, metastatic carcinoid tumours. However, only 70% of cases are MIBG-avid and tumour uptake is not always sufficient for therapy. At The Netherlands Cancer Institute 34 carcinoid patients with no or insufficient uptake were treated with escalating doses of unlabelled ('cold') MIBG. No objective remissions were recorded, but a palliative effect (i.e. subjective disappearance of symptoms and/or reduction of medication by more than 50%) was observed in 60% of cases (mean duration 4.5 months). In 24 of the patients undergoing therapy with 'cold' MIBG, total body scintigraphy using 37 MBq 131I MIBG was performed before and after infusion of 'cold' MIBG. The biodistribution of 131I-MIBG and its tumour to non-tumour ratios were compared. After 'cold' MIBG the 131I-MIBG uptake in the salivary glands was suppressed in all patients, myocardial uptake in 21, and uptake in normal liver tissue in 14. Pulmonary uptake was increased in 13 patients. More importantly, the tumour to non-tumour (T/NT) ratios improved in 17 of the 24 cases (by 7.8-111.4% at 24 h). Of the initial six patients demonstrating a significant increase in the T/NT ratio, five have subsequently received combined treatment of 7.4 GBq 131I-MIBG following the administration of 'cold' MIBG (both by 4 h intravenous infusion), resulting in a good palliative response in four of them. These patients had previously been excluded from therapy with 131I-MIBG only. It is concluded that the administration of unlabelled MIBG may not only provide palliation to patients with carcinoid tumours, but may also alter the biodistribution of MIBG, enabling 131I-MIBG therapy to be used in cases not qualifying for this treatment due to insufficient tumour uptake. PMID- 11039460 TI - Basal and activational 99Tcm-HMPAO brain SPECT in Alzheimer's disease. AB - Early diagnosis in Alzheimer's disease (AD) is important for the administration of new treatments. The purpose of this study was to differentiate mildly/moderately demented AD patients from normal controls by means of activational brain SPECT, and to investigate the correlation between regional cerebral blood flow and dementia severity. Activational brain SPECT was performed 1 week after basal brain SPECT in 12 mild/moderate AD patients according to NINCDS-ADRDA criteria (mean age 69+/-7 years) and in seven healthy, age-matched, volunteer controls (mean age 65+/-9 years). In order to activate the parietal cortex, patients were asked to subtract serial 5's from 100, 2 min before and after the intravenous administration of 925 MBq technetium-99m labelled D,L hexamethyl-propylene amine oxime (99Tcm-HMPAO). Using a three-headed gamma camera equipped with high resolution collimators, 128 images of 35 s duration in a 64 x 64 matrix were obtained over 360 degrees. Region to whole brain ratios (R/WB) were calculated in three consecutive transaxial slices 2 pixels thick above the orbitomeatal line, and the activation percentage was calculated. No statistical difference was detected between AD patients and normal controls for parietal cortex activation. The correlation coefficient between the Mini-Mental State Examination (MMSE) scoring and the activation percentage was 0.475 in normal controls and 0.175 in AD patients for the left anterior parietal cortex, and 0.353 in normal controls and 0.146 in AD patients for the right anterior parietal cortex. In a visual evaluation of parietal cortex activation, 50% of AD patients were able to activate the parietal cortex, whereas 86% of the normal controls could do so. In our current study, the subtraction of serial 5's was not regarded as a promising task. Further studies are needed to clarify the importance of such tasks in the differential diagnosis of mild/moderate AD patients from normal elderly. PMID- 11039461 TI - Brain SPECT findings in long-term inhalant abuse. AB - This study evaluates brain perfusion in long-term inhalant abusers of toluene, acetone, benzene and derivatives. Ten patients in the age range 16-18 years (mean, 17.3+/-0.67 years), who had been inhalant dependent for a mean period of 48.3+/-6.2 months, but who had stopped using inhalants for 1-11 months (mean, 5.4+/-2.1 months), and ten controls (mean age, 17.3+/-0.67 years) were included in the study. Psychiatric tests, biochemical tests and Tc-99m hexamethylpropyleneamine oxime (Tc-99m-HMPAO) brain single photon emission computed tomography (SPECT) were performed on all patients. Brain SPECT images were evaluated qualitatively and quantitatively. The mean IQ level was found to be 84 (by psychological tests). Brain SPECT showed non-homogeneous Tc-99m-HMPAO uptake and hypoperfusion areas in all patients (five left temporal, one right temporal, two left temporal plus bilateral parietooccipital, one biparietal and one left temporoparietal). Seven patients had hyperperfused foci (unifocal in five patients and multifocal in two patients). Six hyperperfused foci were in a parietal and one in a temporoparietal location. This study suggests that inhalant dependents exhibit serious abnormalities in brain SPECT images, including hypo hyperperfusion foci and non-homogeneous uptake of the radiopharmaceutical. A further study with a larger number of patients and long-term follow-up may help to reach a more specific conclusion. PMID- 11039462 TI - The role of hepatobiliary scintigraphy in the evaluation of the protective effects of dimethylsulphoxide in ischaemic/reperfusion injury of liver. AB - Liver ischaemia may lead to parenchymal damage depending on the duration of the ischaemia. Dimethylsulphoxide (DMSO), a well-known radical oxygen scavenger, is a protective agent against ischaemia/reperfusion injury. In this study we aimed to investigate the role of hepatobiliary scintigraphy (HBSc) in detecting the protective effect of DMSO. Eighteen rabbits, in three groups of six, were injected with 37 MBq technetium-99m-mebrofenin via the ear veins. Dynamic scintigrams were taken for 60 min (1 frame/min). In group A, HBSc was performed without any surgery. In groups B and C the Pringle manoeuvre (PM) was applied for 30 min, and tissue specimens for electron microscopy were taken from the liver parenchyma 5 min after the end of the PM. In addition, in group C 1 g/kg DMSO was injected into each rabbit 5 min before application of the PM. HBSc was then performed in groups B and C. From the dynamic images time-activity curves (TACs) were obtained for each group, and the time of peak uptake (TPU) and time for half of the activity to clear from the liver (T1/2) were calculated. The TPU and T1/2 of group B were significantly longer than those of groups A and C (P<0.0005 and P<0.005 for TPU, and P<0.0005 and P<0.02 for T1/2, respectively). The TPU and T1/2 of group C were significantly longer than those of group A (P < 0.005 and P < 0.02, respectively). While the electron microscopic images in group C showed reversible changes, those in group B showed both irreversible and reversible changes. The electron microscopic findings of groups B and C confirmed the scintigraphic findings. In conclusion, HBSc might be used as a practical quantitative method for detecting the protective effects of DMSO. However, its clinical value should be evaluated by further studies with human subjects. PMID- 11039463 TI - Should nuclear medicine physicians give the results of radioisotope examinations directly to patients? AB - Patients often ask for the results after a radioisotope procedure, which can make nuclear medicine physicians feel uncomfortable. In Belgium, nuclear medicine physicians are not supposed to disclose results directly to patients, but to send them to the referring physician. We undertook this work to determine the official rules and practical attitudes in other countries. An introductory letter and a questionnaire were sent to 103 eminent nuclear medicine specialists from 37 countries. Seventy responses (32 countries) were received. Official rules seemed to exist in only seven countries. Most of the respondents indicated that their attitude depended on the clinical situation and the results of the test. Many respondents emphasized that, while in some situations the results should be communicated directly to patients in order to initiate treatment rapidly, in other situations, such as cancer, the referring physician was better suited to disclose the results. The advantages and drawbacks of different attitudes are discussed. Practically and universally applicable rules are difficult to establish, but choosing one solution remains preferable to no standardized attitude at all. An official body, including the medical community, representatives of the population and legal experts, should define an official rule which should be widely communicated, stressing its advantages and drawbacks. In practice, all nuclear medicine physicians would have to do would be to stick to the rule. PMID- 11039465 TI - An immunohistochemical analysis of p27 expression in human pancreatic carcinomas. AB - p27 is known as one of the candidate tumor suppressor genes. Although abnormalities including deletion and mutation in this gene are rarely detected, loss of p27 expression has been reported to be correlated with the high degree of malignancy in many human cancers. In the present study, we investigated the status of p27 expression in human pancreatic carcinoma (PC) and assessed the clinicopathologic significance of loss of p27 expression in the development and progression of this malignancy using immunohistochemistry. No nuclear staining for p27 protein existed in 65 (65.7%) of the 99 PC tumors examined. Furthermore, loss of p27 expression was correlated with tumor grade (G1 versus G2 or G3, p < 0.05) or clinical stage (I and II versus III and IV, p < 0.01). The above data suggest that loss of p27 expression is a very common event in PC, and moreover, this alteration might contribute to the progression of this malignant disease. Key Words: Pancreatic carcinoma-p27 protein expression-Immunohistochemistry. PMID- 11039464 TI - GP2, a GPI-anchored protein in the apical plasma membrane of the pancreatic acinar cell, co-immunoprecipitates with src kinases and caveolin. AB - We previously showed that endocytosis at the apical plasma membrane (APM) of the pancreatic acinar cell is activated by the cleavage of GP2, a GPI-linked protein, from the apical cell surface. This endocytic process, as measured by horseradish peroxidase uptake into pancreatic acinar cells, is blocked by the tyrosine kinase inhibitors genistein and tyrphostin B42 as well as by disruption of actin filaments with cytochalasin. This suggests that the cleavage of GP2 from the cell membrane may activate endocytosis through a tyrosine kinase-regulated pathway. However, the mechanism by which GP2 and tyrosine kinases act together to activate endocytosis at the APM remains unknown. In this study, we demonstrate that pp60, p62yes, caveolin, and annexin, which have previously been implicated in endocytosis in other cell lines, were present in high abundance in GPI-enriched membranes by Western blot analysis. pp60, p62yes, and caveolin all co immunoprecipitated with GP2 except annexin. An 85-kDa protein whose tyrosine dependent phosphorylation is correlated with the activation of endocytosis in intact acinar cells also was present in these immunoprecipitates. This suggests that in pancreatic acini, GP2 may exist in a complex with src kinases, caveolin, and an 85-kDa phosphorylated substrate to regulate endocytosis at the APM. PMID- 11039466 TI - Involvement of calmodulin and protein kinase C in cholecystokinin release by bombesin from STC-1 cells. AB - The mouse intestinal neuroendocrine tumor cell line STC-1 secretes cholecystokinin (CCK) and other hormones. We investigated the role of Ca2+, calmodulin (CaM), and protein kinase C (PKC) in the regulation of CCK release from STC-1 cells. Phorbol 12-myristate 13-acetate (TPA) significantly stimulated CCK release. Staurosporine significantly inhibited CCK release from STC-1 cells stimulated by TPA in a dose-dependent manner. The absence of extracellular calcium completely inhibited CCK release from TPA-stimulated STC-1 cells. Neurotensin did not stimulate CCK release from these cells. W-7, a CaM antagonist, reduced CCK release from STC-1 cells stimulated by bombesin in a dose dependent manner. These findings suggest that CaM and PKC play an important role in the regulation of CCK release from STC-1 cells stimulated by bombesin. PMID- 11039467 TI - Expression of the multidrug-resistance 1 (MDR1) gene and prognosis in human pancreatic cancer. AB - Multidrug-resistance 1 (MDR1) encodes a 170 kDa transmembrane glycoprotein (P glycoprotein), which acts as a drug-efflux pump. In the present study, we analyzed the expression of MDR1/P-glycoprotein in human pancreatic cancer and correlated the results with clinical parameters. Pancreatic cancer tissue samples were obtained from 67 patients (30 female, 37 male) who underwent surgery. Normal pancreatic tissues obtained from 15 previously healthy organ donors (4 female, 11 male) served as controls. MDR1 mRNA levels were analyzed by Northern blotting, and the exact site of MDR1 mRNA expression was determined by in situ hybridization and immunohistochemistry. Northern blot analysis indicated that in comparison with the normal pancreas, MDR1 mRNA levels were only increased 1.4 fold (p = 0.03) in the pancreatic cancer samples. However, there was a 2.9-fold (p < 0.01) increase in MDR1 mRNA levels when only the samples that exhibited increased expression (38%) were analyzed. In situ hybridization and immunohistochemical analysis showed that MDR1 was highly expressed in the cancer cells of these samples. Statistical analysis revealed that patients with high MDR1/P-glycoprotein expression had a shorter postoperative survival time compared with patients with weak to moderate expression of MDR1. On the basis of in situ hybridization, survival in the intense group was 11.6 (n = 12) versus 14.2 months (n = 42) in the mild to moderate group. On the basis of immunohistochemistry, survival in the intense group was 7.5 months (n = 10) versus 14.1 months (n = 40) in the mild to moderate group. Surprisingly, survival of patients with high expression of MDR1/P-glycoprotein was not significantly different from that of patients without detectable MDR1/P-glycoprotein expression. These findings suggest that both strong expression of MDR1/P-glycoprotein and lack of expression seem to influence tumor growth via known and yet unknown mechanisms. PMID- 11039468 TI - Acute pancreatitis results in induction of heat shock proteins 70 and 27 and heat shock factor-1. AB - Heat shock proteins (HSPs) 70 and 27 are stress-responsive proteins that are important for cell survival after injury; the expression of these HSPs is regulated primarily by the transcription factor heat shock factor-1 (HSF-1). The purpose of this study was to determine the effect of acute pancreatitis on pancreatic HSPs (70, 27, 60, and 90) expression and to assess potential mechanisms for HSP induction using a murine model of cerulein-induced pancreatitis. We found an increase of both HSP70 and HSP27 levels with expression noted throughout the pancreas after induction of pancreatitis. HSP60 and HSP90 levels were constitutively expressed in the pancreas and did not significantly change with acute pancreatitis. HSF-1 DNA binding activity increased in accordance with increased HSP expression. We conclude that acute pancreatitis results in a marked increase in the expression of HSP70 and HSP27. Furthermore, the induction of HSP70 and HSP27 expression was associated with a concomitant increase in HSF-1 binding activity. The increased expression of both HSP70 and HSP27 noted with pancreatic inflammation may confer a protective effect for the remaining acini after acute pancreatitis. PMID- 11039469 TI - Calcium concentration and artificial precipitates in human pancreatic juice. AB - We studied the role of the increase in the calcium concentration in pure pancreatic juice of alcoholic noncalcified chronic pancreatitis. Pure pancreatic juice was obtained endoscopically. The pancreatic juice from patients with chronic pancreatitis was adjusted to pH 7.5; then the calcium concentration was adjusted to 0.4, 2.9, 5.4, or 10.4 mmol/L. Artificial precipitates were produced by incubation of the samples at 37 degrees C for 6 hours. Proteins in the artificial precipitates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and the protein patterns were compared with the patterns of natural protein plugs from patients with chronic pancreatitis. The amount of the precipitate increased as the added calcium increased. The protein patterns of SDS-PAGE of the artificial precipitates were similar to those of protein plugs. Albumin, a-amylase, lipase, trypsinogen, and chymotrypsinogen were identified by immunoblotting both in the precipitate and in the protein plug. The increased calcium concentrations in pancreatic juice induced the formation of precipitates whose protein composition was similar to that of protein plugs. An increased calcium concentration in human pancreatic juice may play an important role in the pathogenesis of protein plugs. PMID- 11039470 TI - Synthetic porcine secretin is highly accurate in pancreatic function testing in individuals with chronic pancreatitis. AB - The secretin stimulation test is the most sensitive and specific test for pancreatic function. It is usually performed using biologically derived porcine secretin. Several shortages of biologic porcine secretin have occurred in the past few years. The aim of this study was to compare synthetic porcine secretin to biologic porcine secretin in pancreatic function testing in subjects with chronic pancreatitis. Twelve patients with a previously abnormal secretin stimulation test were enrolled. After an overnight fast, each patient underwent a secretin stimulation test on 2 consecutive days using 1 CU/kg biologic porcine secretin or 0.2 [microg/kg synthetic porcine secretin in a randomized fashion. The peak bicarbonate concentration in duodenal juice was used as a measure of pancreatic function. The peak bicarbonate concentration (mean +/- SD) obtained by using biologic porcine secretin and synthetic porcine secretin were 70 +/- 25 mEq/L and 68 +/- 31 mEq/L, respectively (p = 0.58, paired t test; R = 0.964). The accuracy of synthetic porcine secretin in diagnosing pancreatic insufficiency was 100% when compared with biologic porcine secretin. We conclude that synthetic porcine secretin is highly accurate and safe in pancreatic function testing. The 100% purity, excellent diagnostic accuracy, and ready availability make synthetic porcine secretin an attractive choice for secretin stimulation testing. PMID- 11039471 TI - Severe acute pancreatitis: prognostic factors in 270 consecutive patients. AB - Acute pancreatitis (AP) is a common abdominal disorder with severity varying from mild to fatal disease. Predicting a patient's outcome remains problematic. The aim of this study was to analyze a large consecutive series of patients with severe AP and to identify prognostic factors for hospital mortality. Between 1989 and 1997, a consecutive series of 270 patients with severe AP were included in the study. All patients fulfilled the criteria of Atlanta classification for severe AP. Retrospectively and prospectively collected data included age, gender, etiology, number of previous episodes of pancreatitis, medication history, type of admission, body-mass index (BMI), respiratory failure, renal failure, need for pressor support, and abdominal surgery performed during hospitalization. The overall mortality rate was 24.4%. In univariate survival analysis advanced age, history of continuous medication, patient transferred from other hospital, high BMI, respiratory or renal failure, need for pressor support, and need for abdominal surgery were significant prognostic factors for hospital mortality. In a multivariate stepwise logistic regression analysis, the need of pressor support, renal failure requiring dialysis, advanced age, history of continuous medication and need for abdominal surgery were identified as independent prognostic factors for mortality. A logistic regression analysis of variables available on admission (the first seven above mentioned variables) showed that transferral admission, advanced age, and history of continuous medication were independent prognostic factors for mortality. In patients with severe AP, advanced age, history of continuous medication, and need for dialysis, mechanical ventilator support, and pressor support predict fatal outcome and thus should be taken into account in clinical evaluation. PMID- 11039472 TI - Cigarette smoke enhances ethanol-induced pancreatic injury. AB - Alcohol induces pancreatic ischemia, but the mechanisms promoting pancreatic inflammation are unclear. We investigated whether cigarette smoke inhalation is a cofactor in the development of ethanol-induced pancreatic injury. Cigarette smoke was administered to anesthetized rats alone or in combination with intravenous ethanol infusion. Control animals received either saline or ethanol alone. Pancreatic capillary blood flow and leukocyte-endothelium interaction in postcapillary venules were evaluated by intravital microscopy. Leukocyte sequestration was assessed by measurement of myeloperoxidase activity in pancreatic tissue, and pancreatic injury evaluated by histology. Ethanol decreased pancreatic blood flow progressively over 90 minutes (p < 0.001 vs. baseline), but neither leukocyte-endothelium interaction nor leukocyte sequestration was altered. Cigarette smoke alone reduced pancreatic blood flow temporarily (p < 0.01 vs. baseline) and increased leukocyte-endothelium interaction (roller p < 0.001, sticker p < 0.01 vs. baseline). Cigarette smoke potentiated the impairment of pancreatic capillary perfusion caused by ethanol, and both the number of rolling leukocytes and myeloperoxidase activity levels were increased compared to ethanol or nicotine administration alone (p < or = 0.05 and p < or = 0.01, respectively). This study demonstrates that ethanol induces pancreatic ischemia and that cigarette smoke leads to both temporary pancreatic ischemia and minimal leukocyte sequestration. Cigarette smoke potentiates the amount of pancreatic injury generated by ethanol alone. Smoking therefore seems to be a contributing factor in the development of alcohol-induced pancreatitis in the rat model. PMID- 11039473 TI - Therapeutic efficacy of continuous arterial infusion of an antibiotic and a protease inhibitor via the superior mesenteric artery for acute pancreatitis in an animal model. AB - The major cause of death in acute pancreatitis is severe infection owing to bacterial translocation. As a new strategy, we investigated the effects of continuous intra-arterial infusion of an antibiotic (imipenem) or protease inhibitor (nafamostat mesylate) via the superior mesenteric artery (SMA) on bacterial translocation in acute pancreatitis. Infusion of saline (group I), nafamostat mesylate (group II), or imipenem (group III) was started 6 hours after inducing acute pancreatitis in dogs by infusing autologous gallbladder bile into the main pancreatic duct. The survival rate in group III was significantly improved compared to group I(100 vs. 30% at 24 hours), and bacterial infection of the peritoneal fluid, mesenteric lymph nodes, and pancreas was completely prevented in group III. Intestinal damage assessed by light and scanning electron microscopy and by biochemical parameters (mucosal protein content and myeloperoxidase activity) was also significantly mitigated in group III, which showed milder pancreatic necrosis as well. There was little beneficial effect in preventing bacterial translocation in group II, although the survival rate at 24 hours (70%) was improved. Continuous arterial infusion of an antibiotic via the SMA is effective in mitigating intestinal mucosal damage and preventing bacterial translocation in acute pancreatitis, thereby improving survival. PMID- 11039474 TI - Long-term effects of nafamostat and imipenem on experimental acute pancreatitis in rats. AB - Long-term effects of nafamostat mesylate, a protease inhibitor, and imipenem, an antibiotic, on trypsintaurocholate-induced acute pancreatitis were studied in rats. Sham-operated rats infused with a buffer solution into the pancreatic duct served as controls. Nafamostat (1 mg/kg), imipenem (10 mg/kg), or imipenem + nafamostat in saline was injected subcutaneously 0.25, 3, 24, and 48 hours after the induction of pancreatitis. In untreated rats and control rats, saline was injected at the same intervals as in the treated rats. All rats in an untreated group died within 3.5 days (median survival, 1.25 day) after the induction of pancreatitis. The 2-week survival rate was significantly (p < 0.05) improved by a combination of nafamostat and imipenem (42%), but not by nafamostat (17%), or imipenem (8%) alone. Bacterial culture at 24 hours revealed infection of necrotic pancreatic tissues and ascites by intestinal bacteria in all untreated rats but not in control rats. Bacterial counts were significantly reduced by imipenem, but not by nafamostat. In conclusion, bacterial infection occurred within 24 hours after the induction of trypsintaurocholate pancreatitis in rats. Early treatment with nafamostat + imipenem, but not nafamostat or imipenem alone, improves long term survival. PMID- 11039475 TI - Apoptosis of acinar cells is involved in chronic pancreatitis in Wbn/Kob rats: role of glucocorticoids. AB - The involvement of pancreatic acinar cell apoptosis and its relation to glucocorticoid exposure were investigated in spontaneously occurring chronic pancreatitis in male Wistar Bonn/Kobori (WBN/Kob) rats. Although most lobules were not inflamed in 10-week-old WBN/Kob, increased apoptosis of pancreatic acinar cells, confirmed by TUNEL staining was focally observed (0.10 +/- 0.10 vs. 0.05 +/- 0.10/field in 10-week Wistar rats). Localized hemorrhagic lesions and brown foci in the splenic lobes were apparent, with significant decrease in pancreas weight in 20-week WBN/Kob rats along with marked apoptosis (1.95 +/- 0.31 vs. 0.07 +/- 0.04/field in 20-week Wistar rats). Electron microscopy revealed apoptotic bodies to be present in acinar cells. Pancreatic myeloperoxidase activities, indirect indices of granulocyte infiltration, as well as histologic scores were significantly increased at 15 and 20 weeks, and endogenous corticosterone levels were significantly decreased at 10, 15, and 20 weeks as compared with values for age-matched Wistar rats. Prednisolone in the drinking water (0.01 mg/mL; calculated dose, 1.03 0.03 mg/kg/d) for 10 weeks significantly attenuated increases in numbers of apoptotic acinar cells and pancreatic myeloperoxidase activities and tended to reduce the histologic scores in 20-week WBN/Kob rats as compared with the vehicle group. In summary, (a) apoptosis of pancreatic acinar cells is involved in chronic pancreatitis, (b) endogenous corticosterone is decreased, and (c) prednisolone treatment attenuates both apoptosis of pancreatic acinar cells and chronic pancreatitis in male WBN/Kob rats. We conclude that apoptosis of acinar cells related to decreased corticosterone may be a trigger of chronic pancreatitis in this model. PMID- 11039476 TI - Superiority of mild hypothermic (20 degrees C) preservation for pancreatic microvasculature using the two-layer storage method. AB - Hypothermia causes vascular endothelial damage that leads to graft microcirculation disorder and eventually thrombosis after reperfusion. The two layer cold storage method (TL) was previously demonstrated to supply oxygen to the pancreas graft and maintain high adenosine triphosphate tissue concentration. In this study, we evaluated whether mild hypothermic (20 degrees C) preservation using the TL method could reduce endothelial damage while maintaining parenchymal viability. Graft survival by 20 degrees C preservation was investigated using a dog segmental pancreas autotransplantation model (simple storage in University of Wisconsin solution (UW) for 5 and 8 hours or TL for 5, 8, 12, and 24 hrs. respectively). Subsequently, the grafts were preserved in four different conditions (4 and 20 degrees C UW. 4 and 20 degrees C TL) for 8 hours to evaluate microvascular endothelial damage. Trypan blue uptake of vascular endothelium and pancreatic tissue perfusion were evaluated. No graft preserved by 20 degrees C UW for 5 and 8 hours survived (0/7 and 0/4). In contrast, the graft survival rates by 20 degrees C TL for 5, 8, 12, and 24 hours were 100% (5/5), 80% (4/5), 20% (1/5), and 0% (0/4), respectively. In trypan blue uptake analysis, there were significant differences between 4 and 20 degrees C in both UW and TL (4 degrees C UW, 37% [n = 5) vs. 20 degrees C UW, 13% [n = 4] [p < 0.01]; 4 degrees C TL, 29% [n = 5] vs. 20 degrees C TL, 10% [n = 5] [p < 0.011). The perfusion values in 20 degrees C TL were significantly higher than those in other groups at least for up to 120 minutes after reperfusion (p < 0.01 ). In short-term pancreas preservation, mild hypothermic TL reduced vascular endothelial cell damage and ameliorated graft microcirculation while maintaining parenchymal viability. Mild hypothermic TL may lessen vascular complications in clinical pancreas transplantation when used for several-hour preservation. PMID- 11039478 TI - Pancreatic inflammation, apoptosis, and growth: sequential events after partial pancreatectomy in pigs. AB - Positive signs of pancreas regeneration were observed in rats after induced pancreatitis and partial pancreatectomy (1,2). Although the human pancreas did not regenerate after partial anatomic resection (3), the pig pancreas exhibited growth responses to bombesin after partial pancreatectomy (4). This study was undertaken to establish the time course of pancreatic inflammation, apoptosis, and hypertrophy and/or hyperplasia after partial pancreatectomy in pigs. PMID- 11039477 TI - Physiological concentrations of insulin augment pancreatic cancer cell proliferation and glucose utilization by activating MAP kinase, PI3 kinase and enhancing GLUT-1 expression. AB - Pancreatic carcinoma is characterized by poor prognosis and lack of response to conventional therapy for reasons that are not clear. Because of the structural relationship between the exocrine and endocrine pancreas and high concentrations of islet hormones bathing pancreatic tissue, we hypothesized that pancreatic cancer cell proliferation and glucose utilization are regulated by pancreatic islet hormones, particularly insulin. Based on this, the effect of islet hormones on pancreatic cancer cells in vitro was investigated. Five pancreatic cancer cell lines, CD11, CD18, HPAF, PANC-1, and MiaPaCa2 were used to investigate the effect of islet hormones on cell proliferation, glucose utilization, and GLUT-1 expression. Insulin, but not somatostatin and glucagon, induced pancreatic cancer cell growth in a concentration- and time-dependent manner. At concentrations within the range of those in the intrapancreatic vasculature, insulin (10(-10) 10(-8) mol/L) markedly increased [3H]-thymidine incorporation. Insulin significantly enhanced glucose utilization of pancreatic cancer cells before it enhanced cell proliferation. The MAPK kinase inhibitor PD 098059 abolished insulin-stimulated DNA synthesis and partially reduced insulin-stimulated glucose uptake. In contrast, the PI3 kinase inhibitor wortmannin substantially inhibited insulin-induced glucose uptake and partially blocked thymidine incorporation. Furthermore, after 24-hour treatment with insulin, GLUT-I expression in pancreatic cancer cells was markedly increased, indicating that insulin enhances glucose utilization partly through increasing glucose transport. These findings suggest that insulin stimulates proliferation and glucose utilization in pancreatic cancer cells by two distinct pathways. Insulin augments DNA synthesis mainly by MAP kinase activation and glucose uptake mainly by PI3 kinase activation and enhancement of GLUT-I expression. High intrapancreatic concentrations of insulin are likely to play an important role in stimulating pancreatic cancer growth indirectly by increasing substrate availability as well as by direct action as a trophic factor. PMID- 11039479 TI - The cholecystokinin antagonist proglumide has an analgesic effect in chronic pancreatitis. PMID- 11039480 TI - Acinar Muscarinic Receptors: To Which Subtype Do They Belong? PMID- 11039481 TI - Response to case report: intraductal pancreatic adenoma. PMID- 11039482 TI - Mixing location-relevant and location-irrelevant choice-reaction tasks: influences of location mapping on the Simon effect. AB - The Simon effect refers to the finding that reaction times are faster when stimulus and response locations correspond than when they do not in tasks where stimulus location is defined as irrelevant. The authors examined the Simon effect for situations in which location-irrelevant trials were intermixed with trials for which stimulus location was relevant. Compatible mapping of the location relevant trials enhanced the Simon effect relative to an unmixed condition, whereas incompatible mapping reversed the Simon effect. The reversal with incompatible mapping remained evident when task uncertainty was removed by use of a precue and was larger than the reversed effect produced by making incongruent trials more frequent than congruent trials. This result suggests that both attentional biases and task-defined associations contribute to the reversal of the Simon effect. PMID- 11039483 TI - Spatial attention: different mechanisms for central and peripheral temporal precues? AB - The external noise paradigm (Z.-L. Lu & B. A. Dosher, 1998) was applied to investigate mechanisms of spatial attention in location precuing. Observers were precued or simultaneously cued to identify 1 of 4 pseudocharacters embedded in various amounts of external noise. The cues were either central or peripheral. Both central and peripheral precuing significantly reduced threshold in the presence of high external noise (16% and 17.5%). Only peripheral precuing significantly reduced threshold in the presence of low, or no, external noise (11%). A perceptual template model identified different mechanisms of attention for central and peripheral precuing, external noise exclusion for central precuing, and a combination of external noise exclusion and stimulus enhancement (or equivalently, internal additive noise reduction) for peripheral cuing. PMID- 11039484 TI - An exemplar-retrieval model of speeded same--different judgments. AB - R. M. Nosofsky and T. J. Palmeri's (1997) exemplar-based random-walk (EBRW) model of speeded classification is extended to account for speeded same--different judgments among integral-dimension stimuli. According to the model, an important component process of same--different judgments is that people store individual examples of experienced same and different pairs of objects in memory. These exemplar pairs are retrieved from memory on the basis of how similar they are to a currently presented pair of objects. The retrieved pairs drive a random-walk process for making same--different decisions. The EBRW predicts correctly that same responses are faster for objects lying in isolated than in dense regions of similarity space. The model also predicts correctly effects of same-identity versus same-category instructions and is sensitive to observers' past experiences with specific same and different pairs of objects. PMID- 11039485 TI - The role of talker-specific information in word segmentation by infants. AB - Infants' representations of the sound patterns of words were explored by examining the effects of talker variability on the recognition of words in fluent speech. Infants were familiarized with isolated words (e.g., cup and dog) from 1 talker and then heard 4 passages produced by another talker, 2 of which included the familiarized words. At 7.5 months of age, infants attended longer to passages with the familiar words for materials produced by 2 female talkers or 2 male talkers but not for materials by a male and a female talker. These findings suggest a strong role for talker-voice similarity in infants' ability to generalize word tokens. By 10.5 months, infants could generalize different instances of the same word across talkers of the opposite sex. One implication of the present results is that infants' initial representations of the sound structure of words not only include phonetic information but also indexical properties relating to the vocal characteristics of particular talkers. PMID- 11039486 TI - Sound enhances visual perception: cross-modal effects of auditory organization on vision. AB - Six experiments demonstrated cross-modal influences from the auditory modality on the visual modality at an early level of perceptual organization. Participants had to detect a visual target in a rapidly changing sequence of visual distractors. A high tone embedded in a sequence of low tones improved detection of a synchronously presented visual target (Experiment 1), but the effect disappeared when the high tone was presented before the target (Experiment 2). Rhythmically based or order-based anticipation was unlikely to account for the effect because the improvement was unaffected by whether there was jitter (Experiment 3) or a random number of distractors between successive targets (Experiment 4). The facilitatory effect was greatly reduced when the tone was less abrupt and part of a melody (Experiments 5 and 6). These results show that perceptual organization in the auditory modality can have an effect on perceptibility in the visual modality. PMID- 11039487 TI - Probing distractor inhibition in visual search: inhibition of return. AB - The role of inhibition of return (IOR) in serial visual search was reinvestigated using R. Klein's (1988) paradigm of a search task followed by a probe-detection task. Probes were presented at either the location of a potentially inhibited search distractor or an empty location. No evidence of IOR was obtained when the search objects were removed after the search-task response. But when the search objects remained on, a pattern of effects similar to Klein's results emerged. However, when just the search-critical object parts were removed or when participants received immediate error feedback to prevent rechecking of the search objects, IOR effects were observed only when probes appeared equally likely at search array and empty locations. These results support the operation of object-based IOR in serial visual search, with IOR demonstrable only when rechecking is prevented (facilitating task switching) and monitoring for probes is not biased toward search objects. PMID- 11039488 TI - Sequential effects in number comparison. AB - The modular framework of number processing (e.g., S. Dehaene & R. Akhavein, 1995) was applied to study sequential trial-to-trial effects in a number comparison task. In Experiment 1, numbers were always presented as digits. Responses were faster when the same number was repeated, but this effect was additive with the numerical distance effect. In Experiment 2, numbers were presented either as digits or as words. The authors found significant effects of repeating (a) the same physical stimulus, (b) the same number but in a different notation, and (c) the same notation but a different number. Again, all 3 effects were additive with the numerical distance effect. The authors' results provide strong evidence against accounts according to which, on stimulus repetition trials, the comparison stage is bypassed (as proposed by S. Dehaene, 1996), and the results clearly favor an early, precomparison locus of repetition effects. PMID- 11039489 TI - The chronometry of single neuron activity: testing discrete and continuous models of information processing. AB - The authors propose to study information transmission by comparing the effects of experimental factors on reaction time (RT) with the latency of the changes in activity of single-neurons. An experiment was conducted in which a monkey (Macaca mulatta) performed a tactilo-manual 2-choice RT task and the compatibility of the stimulus-response mapping was manipulated. Task-related neurons were recorded in the monkey's primary somesthetic and motor cortices. The changes in activity of 105 of these neurons were classified either as sensory-like or as motor-like. The sensory-like changes occurred before the motor-like ones. The stimulus-response mapping exerted its entire effect on the RT after the sensory-like changes and before the motor-like ones. These findings suggest that the information was transmitted discretely from the processes affected by the mapping to the processes implemented by the motor-like changes. PMID- 11039490 TI - Visual and motor effects in inhibition of return. AB - Inhibition of return (IOR) refers to slowed reaction times (RTs) when a target appears in the same rather than a different location as a preceding stimulus. The present study tested the hypothesis that IOR reflects a motor bias rather than a perceptual deficit. Two signals (S1 and S2) were presented on each trial. These signals were peripheral onsets or central arrows. The responses required to S1 and S2 were, respectively, no response-manual, manual-manual, saccadic-manual, no response-saccadic, manual-saccadic, and saccadic-saccadic. Uniting perceptual and motor bias views of IOR, the results demonstrated inhibition for responding to (a) peripheral signals when the eyes remained fixed (slowed visual processing) and (b) both peripheral and central signals when the eyes moved (slowed motor production). However, the results also emphasized that the nature of IOR depends fundamentally on the response modality used to reveal its influence. PMID- 11039491 TI - Body-scaled transitions in human grip configurations. AB - This article reports two experiments that were set up to examine the preferred human grip configuration used to displace cubes that varied in length (Lc), mass (Mc), and density (ML3). In particular, the authors sought to provide a more precise test of a dimensional relation between the object and the hand that had previously been shown to predict the grip configuration used to transport an object from one location to another. The experiments examined 2 grip transitions (from 3 digits to 4 digits and from 1 hand to 2 hands) within 2 sets of object conditions. In Experiment 1, cubes with a low density and a small increment in size (1 mm) were used, whereas in Experiment 2, cubes with 2 fixed sizes and small increments in mass were used. The results showed that the body-scaled equation K = logLc + (logMc/a + bMh + cLh), where Mh and Lh are the anthropometric measures of the hand mass and length and a, b, and c are empirical constants, is the body-scaled information that predicts the grip configurations used to displace objects. PMID- 11039492 TI - In vitro culture of rat hepatocytes without exogenous matrix. PMID- 11039493 TI - Estrogen mitogenic action. I. Demonstration of estrogen-dependent MTW9/PL2 carcinogen-induced rat mammary tumor cell growth in serum-supplemented culture and technical implications. AB - The MTW9/PL cell line was established by our laboratory in culture from the carcinogen-induced hormone-responsive MT-W9A rat mammary tumor of a Wistar-Furth (W/Fu) rat. This tumor formed estrogen, androgen, and progesterone responsive tumors in W/Fu rats (Sirbasku, D. A., Cancer Res. 38:1154-1165; 1978). It was later used to derive the MTW9/PL2 cell population which was also estrogen responsive in vivo (Danielpour, D., et al., In Vitro Cell. Dev. Biol. 24:42-52; 1988). In the study presented here, we describe serum-supplemented culture conditions in which the MTW9/PL2 cells demonstrate > or = 80-fold steroid hormone growth responses. All sera used were steroid hormone-depleted by charcoal-dextran treatment at 34 degrees C. The studies were done with horse serum as well as serum from other mammalian species. The growth of the MTW9/PL2 cells was biphasic in response to hormone-depleted serum. Concentrations of < or = 5% (v/v) promoted optimum growth. Above this concentration, serum was inhibitory. Concentrations > or = 40% (v/v) inhibited growth altogether. Addition of 1.0 x 10(-13)-1.0 x 10( 8) M 17beta,-estradiol (E2) reversed the inhibition completely. At 1.0 x 10(-8) M, estrone, estriol and diethylstilbestrol promoted growth as well as E2. Testosterone and dihydrotestosterone promoted growth only at > or = 10(-7) M. Progesterone was effective only at > or = 10(-6) M. Cortisol was ineffective. Labeled-hormone-binding analysis and Western immunoblotting documented that MTW9/PL2 cells had estrogen and progesterone receptors but not androgen or cortisol receptors. Estrogen treatment of MTW9/PL2 cells induced a concentration and time dependent increase in progesterone receptors. We conclude (1) the MTW9/PL2 population is the first highly steroid hormone-responsive rat mammary tumor cell line to be established in culture from a carcinogen-induced tumor, and (2) sera from a number of species including horse, rat and human contain an inhibitor which mediates estrogen sensitive MTW9/PL2 cell growth in culture. PMID- 11039496 TI - Reciprocal control of apoptosis and proliferation in cultured rat hepatoma arl-6 cells: roles of nutrient supply, serum, and oxidative stress. AB - In order to understand how cancer cells accumulate, rat hepatoma ARL-6 cells were cultured for 8 d to identify factors involved in spontaneous cell proliferation and apoptosis. With increasing time in culture, the proportion of cells in the proliferative phases of the cell cycle and the rate of deoxyribonucleic acid (DNA) synthesis decreased. The waning of proliferation was associated with a gradual reduction of cell viability, and this was temporally related to the appearance of typical apoptotic morphology and DNA laddering. Medium replacement or supplementation with fetal calf serum (FCS) suppressed apoptosis, while medium change, but not fetal calf serum alone, enhanced cell proliferation. Apoptosis was also suppressed by dimethyl sulfoxide (DMSO), but supplementary glutathione was without effect. Expression of poly(adenosine diphosphate[ADP] ribose)polymerase peaked on days 34 of culture, and was followed by a progressive decrease thereafter, consistent with proteolytic cleavage. This decrease was prevented to varying extents by complete medium replacement, FCS and DMSO, indicating a close temporal relationship between poly(ADP-ribose)polymerase activation and apoptosis. Expression of Fas and Bcl-2 did not change appreciably over the 8-d culture, but there was a gradual increase in Bax expression; medium change, FCS and DMSO all partly inhibited Bax expression. These data indicate that spontaneous apoptosis in cultured ARL-6 cells is inversely related to cell proliferation, and that nutrient supply, and to a lesser extent, serum-derived factors and oxidative stress modulate apoptosis in this system. Proteolytic cleavage of poly(ADP-ribose)polymerase and expression of Bax are likely to be mechanistically involved with the control of spontaneous apoptosis in ARL-6 cells, whereas changes in the levels of Fas and Bcl-2 do not play a role. PMID- 11039495 TI - Estrogen mitogenic action. III. is phenol red a "red herring"? AB - The reported estrogenic action of phenol red and/or its lipophilic contaminants has led to the widespread use of indicator-free culture medium to conduct endocrine studies in vitro. Because we have recently developed methods to measure large-magnitude estrogen effects in the tissue culture medium containing phenol red, we concluded that the indicator issue required further evaluation. To do this, we selected nine estrogen receptor positive (ER+) cell lines representing four target tissues and three species. We investigated phenol red using five different experimental protocols. First, 17beta-estradiol (E2) responsive growth of all nine ER+ cells lines was compared in the medium with and without the indicator. Second, using representative lines we asked if phenol red was mitogenic in the indicator-free medium. The dose-response effects of phenol red were compared directly to those of E2. Third, we asked if tamoxifen-inhibited growth equally in phenol red-containing and indicator-free medium. This study was based on a report indicating that antiestrogen effects should be seen only in phenol red-containing medium. Fourth, we asked if phenol red displaced the binding of 3H-E2 using ERK intact human breast cancer cells. Fifth, we compared E2 and phenol red as inducers of the progesterone receptor using a human breast cancer cell line. All the experiments presented in this report support the conclusion that the concentration of phenol red contaminants in a standard culture medium available today is not sufficient to cause estrogenic effects. In brief, our studies indicate that the real issue of how to demonstrate estrogenic effects in culture resides elsewhere than phenol red. We have found that the demonstration of sex steroid hormone-mitogenic effects in culture depends upon conditions that maximize the effects of a serum-borne inhibitor(s). When the effects of the inhibitor are optimized, the presence or absence of phenol red makes no everyday difference to the demonstration of estrogen mitogenic effects with several target cell types from diverse species. PMID- 11039497 TI - Diffusable growth factors induce bladder smooth muscle differentiation. AB - Bladder smooth muscle differentiation is dependent on the presence of bladder epithelium. Previously, we have shown that direct contact between the epithelium and bladder mesenchyme (BLM) is necessary for this interaction. In this study, we tested the hypothesis that bladder smooth muscle can be induced via diffusable growth factors. Fourteen-day embryonic rat bladders were separated into bladder mesenchyme (prior to smooth muscle differentiation) and epithelium by enzymatic digestion and microdissection. Six in vitro experiments were performed with either direct cellular contact or no contact (1) 14-d embryonic bladder mesenchyme (BLM) alone (control), (Contact) (2) 14-d embryonic bladders intact (control), (3) 14-d embryonic bladder mesenchyme combined with BPH-1 cells (an epithelial prostate cell line) in direct contact, (4) 14-d embryonic bladder mesenchyme with recombined bladder epithelium (BLE) in direct contact, (No Contact) (5) 14-d embryonic bladder mesenchyme with BPH-1 prostatic epithelial cells cocultured in type 1 collagen gel on the bottom of the well, and (6) 14-d embryonic bladder mesenchyme with BPH-1 epithelium cultured in a monolayer on a transwell filter. In each case the bladder tissue was cultured on Millicell-CM 0.4-microm membranes for 7 d in plastic wells using serum free medium. Growth was assessed by observing the size of the bladder organoids in histologic cross section as well as the vertical height obtained in vitro. Immunohistochemical analysis of the tissue explants was performed to assess cellular differentiation with markers for smooth muscle alpha-actin and pancytokeratin to detect epithelial cells. Control (1) bladder mesenchyme grown alone did not exhibit growth or smooth muscle and epithelial differentiation. Contact experiments (2) intact embryonic bladder, (3) embryonic bladder mesenchyme recombined with BPH-1 cells, and (4) embryonic bladder mesenchyme recombined with urothelium each exhibited excellent growth and bladder smooth muscle and epithelial differentiation. Both noncontact experiments (5) and (6) exhibited growth as well as bladder smooth muscle and epithelial differentiation but to a subjectively lesser degree than the contact experiments. Direct contact of the epithelium with bladder mesenchyme provides the optimal environment for growth and smooth muscle differentiation. Smooth muscle growth and differentiation can also occur without direct cell to cell contact and is not specific to urothelium. This data supports the hypothesis that epithelium produces diffusable growth factors that induce bladder smooth muscle. PMID- 11039498 TI - beta-adrenergic receptor population is up-regulated by increased cyclic adenosine monophosphate concentration in chicken skeletal muscle cells in culture. AB - Skeletal muscle hypertrophy is promoted in vivo by administration of beta adrenergic receptor (betaAR) agonists. Chicken skeletal muscle cells were treated with 1 microM isoproterenol, a strong betaAR agonist, between days 7 and 10 in culture. betaAR population increased by approximately 40% during this treatment; however, the ability of the cells to synthesize cyclic adenosine monophosphate (cAMP) was diminished by twofold. Neither the basal concentration of cAMP nor the quantity of myosin heavy chain (MHC) was affected by the 3-d exposure to isoproterenol. To understand further the relationship between intracellular cAMP levels, betaAR population, and muscle protein accumulation, intracellular cAMP levels were artificially elevated by treatment with 0-10 betaM forskolin for 3 d. The basal concentration of cAMP in forskolin-treated cells increased up to sevenfold in a dose-dependent manner. Increasing concentrations of forskolin also led to an increase in betaAR population, with a maximum increase of approximately 40-60% at 10 microM forskolin. A maximum increase of 40-50% in the quantity of MHC was observed at 0.2 microM forskolin, but higher concentrations of forskolin reduced the quantity of MHC back to control levels. At 0.2 microM forskolin, intracellular levels of cAMP were higher by approximately 35%, and the betaAR population was higher by approximately 30%. Neither the number of muscle nuclei fused into myotubes nor the percentage of nuclei in myotubes was affected by forskolin at any of the concentrations studied. PMID- 11039494 TI - Estrogen mitogenic action. ii. negative regulation of the steroid hormone responsive growth of cell lines derived from human and rodent target tissue tumors and conceptual implications. AB - In an accompanying report (Moreno-Cuevas, J. E.; Sirbasku, D. A., In Vitro Cell. Dev. Biol.; 2000), we demonstrated 80-fold estrogen mitogenic effects with MTW9/PL2 rat mammary tumor cells in cultures supplemented with charcoal-dextran treated serum. All sera tested contained an estrogen reversible inhibitor(s). The purpose of this report is to extend those observations to additional sex steroid responsive human and rodent cell lines. Every line tested showed a biphasic response to hormone-depleted serum. Concentrations of < or = 10% (v/v) promoted substantive growth. At higher concentrations, serum was progressively inhibitory. With estrogen receptor-positive (ER+) human breast cancer cells, rat pituitary tumor cells, and Syrian hamster kidney tumor cells, 50% (v/v) serum caused significant inhibition, which was reversed by very low physiologic concentrations of estrogens. This same pattern was observed with the steroid hormone-responsive LNCaP human prostatic carcinoma cells. Because steroid hormone mitogenic effects are now easily demonstrable using our new methods, the identification of positive results has nullified our original endocrine estromedin hypothesis. We also evaluated autocrine/paracrine growth factor models of estrogen-responsive growth. We asked if insulin-like growth factors I and II, insulin, transforming growth factor alpha, or epidermal growth factor substituted for the positive effects of estrogens. Growth factors did not reverse the serum-caused inhibition. We asked also if transforming growth factor beta (TGFP) substituted for the serum-borne inhibitor. TGFbeta did not substitute. Altogether, our results are most consistent with the concept of a unique serum-borne inhibitor as has been proposed in the estrocolyone model. However, the aspect of the estrocolyone model related to steroid hormone mechanism of action requires more evaluation. The effects of sex steroids at picomolar concentrations may reflect mediation via inhibitor "activated" intracellular signaling pathways. PMID- 11039499 TI - Carcinoma of the kidney, testis, and rare urologic malignancies. AB - The purpose of this symposium was to provide a forum for discussion on current information on the etiology and diagnosis of, and therapy for, tumors of the kidney, testis, and several uncommon malignancies of the genitourinary tract. The most recent contributions in epidemiology and molecular genetics were discussed with specific reference to their importance for clinical practice. Contemporary treatment approaches with the emphasis on multidisciplinary patient management of tumors commonly seen in the clinic as well as those that are only rarely diagnosed by urooncologists were presented. Major stress was given to the management optimization as it pertains to short- and long-term quality of life issues of patients treated for these tumors. Methods to reduce treatment toxicity including carcinogenic potential of chemotherapy, radiotherapy, or their combination were found to be of nearly equal importance to patient survival. Symposium participants reached consensus on a number of important points: 1) The management of patients with several malignancies discussed requires the presence of a multidisciplinary team of specialist who are interested in diagnosis and treatment of genitourinary tumors; 2) Patients managed in such an environment are expected to have optimal survival and the best possible quality of life; 3) Real advances in the management of patients can be best obtained through well-designed prospective clinical trials; and 4) There is a need for timely introduction of relevant advances in epidemiology and molecular genetics to clinics. PMID- 11039500 TI - Examining the use of breast-conserving treatment for women with breast cancer in a managed care environment. AB - At a National Institutes of Health Consensus Conference in 1991, conservation treatment was considered preferable for patients with early-stage breast cancer. In the early and mid-1990s, however, less than half of the eligible patients received this treatment and the rates varied with patient and provider characteristics. This study explores whether more eligible patients with breast cancer received conservation treatment in recent years in a managed care environment compared to reports in the literature, and if patient and hospital characteristics affected the rate of acceptance. The study population included 753 women with breast cancer in clinical stages 0, I, or II. Patients with Stage III or IV tumors or with tumors larger that 5.0 cm were excluded. A multiple logistic regression incorporated in a mixed-effect model was used to estimate the effect of patient and facility characteristics on the likelihood of using breast conserving surgery controlling for clinical stages and demographics such as age, race, and marital status. Among the 753 eligible patients, 474 (62.9%) received conservation surgery. Only Hispanic ethnicity and clinical stage significantly affected the likelihood of receiving conservation treatment. Factors such as patient age, hospital size, and teaching status that had been found to be significant predictors in earlier studies were not statistically significant in this study, although conservation treatment was more frequent in younger women and in teaching hospitals. A larger proportion of eligible patients received conservative treatment in this study than in previous reports. This treatment became available in a broader range of institutions, moving from large, academic teaching centers to smaller community hospitals. PMID- 11039501 TI - Combined preoperative chemotherapy and radiation for locally advanced rectal carcinoma. AB - To determine the efficacy of combined preoperative chemotherapy and radiation therapy for locally advanced rectal carcinoma and the rate of sphincter conservation, a retrospective survey of 39 patients with locally advanced rectal carcinoma treated with various 5-fluorouracil- and leukovorin-based chemotherapy regimens and radiation prior to surgery in a single institution was reviewed. Toxicity, local control and survival were evaluated and compared to previous studies with similarly staged patients. Long-term follow-up was available on 35 patients. The actuarial local failure was 5.7% while the actuarial 5-year survival was 87%. The mortality rate was low (2.5%) and the rate of long-term serious complications acceptable (11.4%). Combined preoperative chemotherapy and radiation provided excellent local regional control despite the poor prognostic factors associated with size, fixation, and the initial advanced tumor stage with acceptable morbidity. In addition, patients with tumors located in the lower third of the rectum may be able to undergo sphincter-sparing surgery. Although the median follow-up is relatively short (32.4 months), the results are in accordance with previous studies of neoadjuvant combined chemotherapy and radiation for locally advanced rectal carcinoma in terms of local and distant control. PMID- 11039502 TI - Phase I clinical trial of oral furtulon and combined hepatic arterial chemoembolization and radiotherapy in unresectable primary liver cancers, including clinicopathologic study. AB - Surgical resection has been accepted as the only curative therapy for primary liver cancer (PLC). Unfortunately, most patients are surgically unresectable when they seek treatment. An alternative therapeutic approach for some of these patients is transcatheter arterial chemoembolization. However, this is not curative by itself, and additional therapy is required to eradicate residual disease. This study investigates the approach of preoperative hepatic arterial chemoembolization followed by the combination of oral Furtulon (5'-deoxy-5 fluorouridine) as a radiosensitizer and external beam radiotherapy (RT). From July 1997 to December 1998, 25 patients with unresectable PLC were treated with hepatic arterial chemoembolization followed by limited-field radiotherapy plus oral Furtulon as a radiosensitizer. Hepatic arterial chemoembolization was performed with 5-fluorouracil 1 g, cisplatin 80 mg (DDP), mitomycin C (MMC) 10 mg, and arterial embolization with iodized oil-10 ml mixed with 10 mg MMC. Hepatic arterial chemoembolization was performed at regular intervals of 6 weeks, and the patients then received limited-field RT. Mean tumor dose was 4,600 cGy (range, 4,100-5,200 cGy) in daily 1.8-Gy fractions, 5 times a week. The toxicity and responses between RT and surgery were assessed. After surgical evaluation, resection was performed. The histopathologic study was also performed in the specimens of both normal and radiation-injured liver tissues from the patients who underwent resection. Seventeen of 25 patients (68%) showed an objective response. One patient with cholangiocarcinoma involving the portal lymph nodes attained a complete response. Eight patients (32%) underwent sequential resection. The most common toxicity was an increase in liver enzymes, which were less than twofold of the upper limit of normal. Follow-up computed tomography studies after treatment showed a low-attenuation area adjacent to the hepatic tumor in the target volume. On pathologic evaluation, the low-attenuation area revealed hyperemia, distended hepatic sinusoids packed with erythrocytes, and hepatic cell loss when examined with microscopy; "new-born" hepatocytes, hepatic cords in the process of forming, and endothelial cells have appeared on electronic microscopic examination. The combination of hepatic arterial chemoembolization and external radiotherapy is efficacious and a safe modality for unresectable primary liver cancers. Furtulon offers the potential for use as a clinical radiosensitizer. Radiation can significantly damage the liver tissue between 41 Gy and 52 Gy, but the new hepatocytes were forming within the radiation-injured liver after RT. PMID- 11039503 TI - Small cell carcinoma of the esophagus: analysis of 10 cases and review of the published data. AB - Small cell carcinoma of the esophagus is a rare and aggressive tumor with early widespread dissemination. In this retrospective study, we report epidemiologic, histologic, and clinical characteristics of small cell carcinoma of the esophagus from the analysis of 10 patients, with a literature review. Between 1993 and 1998, 10 patients with small cell carcinoma of the esophagus were treated in our institution, representing 2.8% of all esophageal malignancies diagnosed during this period. Four patients sought treatment for limited disease, whereas six patients had distant metastasis at the time of diagnosis. All patients received polychemotherapy, and a complete response was observed in eight patients. Seven of these patients received subsequent locoregional radiotherapy, with endoesophageal brachytherapy in two patients. The overall median survival was 15.5 months (range, 2-36 months) for all of the patients. In limited stages, the overall median survival was 18.5 months (range, 2-36 months), whereas it was 11 months (range, 6-19 months) for the extensive stage at initial diagnosis. In this article, we report our experience with this uncommon neoplasia and attempt to make comparisons with the cases published in the literature regarding location, symptoms, histopathologic diagnosis, and treatment. We conclude that the optimum treatment seems to be the same as for small cell carcinomas of the lung, that is, a multidrug combination chemotherapy regimen used alone or with sequential radiation. PMID- 11039504 TI - Peritoneal carcinomatosis in germ-cell tumor: relations with retroperitoneal lymph node dissection. AB - Peritoneal carcinomatosis from germ-cell tumor has rarely been described, and thus remains largely unknown. We report here five cases involving this entity. All five patients had embryonal carcinoma in their primary germ-cell tumor. Four of them had undergone retroperitoneal lymph node dissection (RPLND), and viable malignant cells were found. RPLND was performed for relapses (n = 3) and as primary therapy for stage II disease (n = 1). The peritoneum was the only site of relapse in three patients, and was associated with pleural effusion in one. The time to relapse after RPLND ranged from 6 to 14 months. One patient sustained injury to lymph nodes during RPLND, and another patient had a peritoneal xanthelasma. The only three patients already described in the literature underwent RPLND or surgical biopsy. All these observations suggest a striking relation between RPLND and occurrence of subsequent peritoneal carcinomatosis. PMID- 11039505 TI - Cellular immune profile of patients with advanced cancer before and after taxane treatment. AB - The purpose of this study is to determine immune recovery and function after treatment with docetaxel or paclitaxel. Peripheral blood mononuclear cells were harvested before chemotherapy and at weekly times afterwards for cycle 1. Leukocyte subsets ICD45hiCD14lo polymorphonuclear neutrophils, CD45hiCD14hi monocytes, CD45hiCD14- lymphocytes, CD3+CD4/CD8+ T cells, CD3-CD19+ B cells, CD3 CD16/CD56+ natural killer (NK) cells], and circulating cytokine levels [tumor necrosis factor-alpha, gamma-interferon (gamma-IFN), and interleukins (IL-2, IL 10, IL-12)] were followed. In addition, T-cell mitogenic function, NK function, and lymphokine activated killer (LAK) function was assessed. Ten patients were entered in the trial. T-cell frequency, B-cell frequency, and CD4/CD8 ratio did not change. IL-10 serum levels significantly decreased in paclitaxel-treated patients (4.4+/-1.3 pg/ml at week 4 versus 7.8+/-2.1 pg/ml at baseline; p < 0.05). IL-2, IL-12, and gamma-IFN levels were not detectable. NK cytotoxic activity decreased in docetaxel-treated patients. LAK cell activity was not altered. Four patients achieved a partial or complete response. They demonstrated higher than normal CD4:CD8 T-cell ratios and an improved phytohemagglutinin stimulation index (SI = 2.5). In conclusion, our findings suggest that immune function was affected more significantly after docetaxel treatment. Investigational approaches, which enhance cellular immunity, may be of greater relevance after treatment with docetaxel. Additional studies monitoring NK function after chemotherapy are recommended. PMID- 11039506 TI - Late recurrence of small-cell lung cancer: a case report. AB - A 67-year-old man was admitted with small-cell lung cancer (SCLC). The patient was given four courses of platinum-containing chemotherapy followed by chest irradiation, and good partial response (PR) was obtained. The patient did well for 4 years, until he sought treatment for a painful subcutaneous tumor. Chest computed tomography scan revealed the mass extending from the tumor in lung parenchyma with osteolytic lesion of the third rib bone. Pathologic examination of the subcutaneous lesion revealed SCLC. The patient was given two courses of the same combination chemotherapy administered as initial therapy. Regression of the mass was observed, and the response was evaluated as a good PR. How to approach late recurrence of SCLC is discussed. PMID- 11039507 TI - Characteristics and outcome of endometrial carcinoma patients age 45 years and younger. AB - Recent reports have suggested that the pathologic features of young patients with endometrial cancer are less favorable than previously thought. We retrospectively reviewed the characteristics and outcome of young patients with endometrial cancer at our institution. A total of 457 surgically staged patients were divided in 2 groups: Group A (age < or =45 years, n = 41) and B (age >45, n = 416). Groups A and B had a similar distribution of tumor stage, grade, histology, lymphovascular invasion, synchronous ovarian primaries, and positive cytology. Although group A tumors had less myometrial invasion (MI) (p = 0.004) and were lower grade (p = 0.06), a trend to more frequent nodal involvement was seen in group A women (p = 0.09). Adverse pathologic features, in particular deep MI, were more common in group A patients older than age 40. Group A patients had a disease-free (p = 0.56) and cause-specific (p = (0.26) survival that was similar to that of group B patients. Young patients with endometrial cancer have a distribution of most pathologic features and equivalent outcome similar to that of older women. However, adverse features are not equally distributed in young women. A discordance may also exist between MI, grade, and nodal involvement. PMID- 11039509 TI - Comparison of posterior fossa and tumor bed boost in medulloblastoma. AB - To quantify the difference between the area of brain irradiated using the posterior fossa boost (PFB) and tumor bed boost (TBB) in medulloblastoma, we studied 15 simulation radiographs of patients treated in our institution from 1990 and 1999. The PFB was compared with the TBB, which was defined as the tumor bed plus 2-cm margin as demonstrated by postoperative magnetic resonance imaging. The PFB field treated a mean area of 9.43 cm2 more brain than the TBB. In 3 patients (20%), the area of the brain in the TBB was larger than the PFB. In 11 patients (73.3%), the PFB field had more than 10% more brain than the TBB. The cochlea was in the PFB and TBB field in all patients. In more than two thirds of patients, the area of brain irradiated with the PFB was at least 10% greater than the TBB. Future studies are needed to determine whether the TBB can replace the PFB in patients with medulloblastoma. PMID- 11039508 TI - Neoadjuvant chemotherapy with ifosfamide, cisplatin, and vinorelbine in advanced squamous cell carcinoma of the cervix. AB - A phase II trial was performed to assess the efficacy and toxicity of a combination of ifosfamide (IFX), cisplatin (CDDP), and vinorelbine (VNB) as neoadjuvant chemotherapy (NAC) for untreated advanced cervical carcinoma (ACC). Between October 1995 and February 1998, 40 patients were entered in this study. Their median age was 43 years (range: 23-74 years). International Federation of Gynecology and Obstetrics stages were: IIB, 23; IIIB, 13; and IVA, 4. Therapy consisted of: IFX 2,000 mg/m2 1-hour (H) IV infusion days 1 to 3; 2 mercaptoethanesulfonic acid sodium salt (mesna) 400 mg/m2 IV bolus H 0 and 4, and 800 mg/m2 by mouth H 8, days 1 to 3; VNB 25 mg/m2 20-minute IV infusion days 1 and 8; and CDDP 75 mg/m2 IV day 3. Cycles were repeated every 28 days for a total of three courses. Both staging and response (R) assessment were performed by a multidisciplinary team. An objective response (OR) was observed in 24 of 40 patients (60%; 95% confidence interval, 45-75%). Four patients achieved complete response (CR) (10%); 20 partial response (50%); 12 patients stable disease (30%); and 4 progressive disease (10%). Eight of 24 patients (33%) with OR underwent radical surgery, and histologic CRs were recorded in 2 of them. The remaining patients received definitive radiotherapy after NAC. The dose-limiting toxicity was myelosuppression. Leukopenia occurred in 32 patients (80%) and was grade III or IV in 14 patients (36%). Peripheral neuropathy occurred in 9 patients (22%), whereas myalgias occurred in 10 (25%). Constipation was observed in 9 patients (23%); emesis occurred in 35 patients (88%). There were no therapy-related deaths. These results indicate that IFX/CDDP/VNB is an active combination for ACC with moderate toxicity. Implementation of this regimen in a multimodal therapy protocol deserves further study. PMID- 11039510 TI - Acute spontaneous tumor lysis syndrome in adenocarcinoma of the lung: a case report. AB - Acute tumor lysis syndrome (ATLS) is a constellation of metabolic complications that typically occurs in the setting of treatment of hematologic malignancies. On occasion, it has been reported to occur after therapy for solid tumors associated with large tumor burdens and aggressive therapy. We herein report the occurrence of spontaneous acute tumor lysis syndrome in a man with untreated metastatic adenocarcinoma of the lung, and briefly discuss the literature. PMID- 11039511 TI - Radiotherapy for postmastectomy local-regional recurrent breast cancer. AB - From 1980 to 1989, 70 women with postmastectomy local-regional recurrent breast cancer and no clinical or radiographic evidence of distant metastasis were treated with radiotherapy with or without systemic therapy. The interval from mastectomy to local-regional recurrence ranged from 5 to 240 months (median, 34 months). The chest wall alone was involved in 37 patients, the supraclavicular area in 12 patients, the internal mammary node area in 3 patients, the infraclavicular area in 2, and the axilla in 1. Fifteen patients had multiple areas of involvement. The complete response rate was 87%. Further local-regional recurrence developed in at least 21 patients, and distant metastasis developed in at least 41 patients. Twenty-five patients (36%) survived at least 5 years and 15 patients (21%) survived at least 10 years. An initial negative node status and long disease-free interval from mastectomy to recurrence were associated with an improved postrecurrence survival. Patients with local-regional recurrence postmastectomy who do not have clinical or radiographic evidence of distant metastasis should be treated aggressively with radiotherapy with or without systemic therapy. Distant metastasis will develop in most such patients, but the majority will remain free of further local-regional recurrence. PMID- 11039512 TI - Results of breast-conserving therapy for early stage breast cancer: Kyoto University experiences. AB - This study evaluated the results of breast-conserving therapy (BCT). Nine hundred six patients who underwent BCT at our hospital between November 1987 and February 1998 were analyzed. The mean age was 48 years. According to the Union Internationale Contre le Cancer 1997 classification system, stages 0, I, IIA, IIB, IIIA, and IIIB were 37, 400, 344, 117, 7, and 1, respectively. Radiation therapy consisted of 50 Gy to the ipsilateral whole breast. Boost irradiation of 10 Gy was administered to 186 of 231 patients with close or positive margins. Nearly all patients received adjuvant chemohormonal therapy with tamoxifen and 5 fluorouracil or its derivatives for 2 years. The minimum and median follow-up periods were 18 and 52 months, respectively. The 5-year overall survival, cause specific survival, local recurrence-free survival, and disease-free survival rates were 97.3%, 98.4%, 98.1%, and 91.5%, respectively. Local recurrence in preserved breast occurred in 20 patients 7 to 86 months after surgery. Multivariate analysis revealed that the most predictive factor for disease-free survival rates and distant failures was the number of pathologically positive lymph nodes (p < 0.0001), and that the factor for local failure was marginal status (p = 0.005). This study demonstrated that BCT was suitable for the treatment of early-stage breast cancer with its reasonable survival rates and acceptable toxicity. PMID- 11039513 TI - Multiple intraabdominal soft-tissue masses in a man awaiting liver transplantation: a case study and discussion. AB - An unusual cause of abdominal soft-tissue masses is accessory splenic tissue. The Tc-99m-sulfur colloid liver-spleen scan is a valuable adjunct in making this diagnosis. This report describes a 47-year-old man who had an abdominal magnetic resonance imaging (MRI) scan as part of a pretransplant evaluation and was found to have multiple soft-tissue masses in the posterior aspect of his abdomen. His history was pertinent for a traumatic rupture of the spleen at the age of 12 years, for which he required a splenectomy. He had no symptoms or physical findings to indicate a lymphoproliferative disorder or other malignant process. His peripheral blood smear was remarkable for the absence of Howell-Jolly bodies. The nuclear scan confirmed the presence of uptake in the soft-tissue masses seen on MRI scan. This finding supports our diagnosis of splenosis in a man with a history of traumatic splenic rupture as a child. PMID- 11039515 TI - Aggressive radiotherapy adjuvant to peripheral blood stem cell transplant for relapsed Hodgkin's disease. AB - The role of radiotherapy in conjunction with high-dose chemotherapy and autologous bone marrow transplant for relapsed Hodgkin's disease remains to be clearly defined. Although there is substantial evidence that radiotherapy enhances local tumor control, prospective trials in the transplant setting have not been reported, and the potential toxicity of radiotherapy need to be considered. However, certain patients are at high risk of posttransplant tumor recurrence, most notably those with tumors unresponsive to pretransplant chemotherapy. We report the use of aggressive radiotherapy in three high-risk patients, including the first reported case of whole lung irradiation after a high-dose carmustine-based chemotherapy regimen. Two of these patients received repeat partial lung irradiation, including one patient with carmustine-related pulmonary toxicity. Radiotherapy (30-34.5 Gy; 1.5 Gy/fraction) was tolerated well without significant acute or late toxicity, and all patients remain disease free 40 to 62 months after irradiation without severe sequelae. We conclude that radiotherapy may be of benefit for patients at high risk of local tumor relapse, and should be considered in such cases despite potential toxicity. PMID- 11039514 TI - Pilot study of organ preservation multimodality therapy for locally advanced resectable oropharyngeal carcinoma. AB - The purpose of this study was to determine the early efficacy and toxicity of a new multimodality organ-preservation regimen for locally advanced, resectable oropharyngeal squamous cell carcinoma (SCC). Patients with T3-4N0-3M0 or T2N2-3M0 oropharyngeal SCC were eligible for this Phase II study. Patients needed the physiologic reserve for surgery and technically resectable tumors. Induction carboplatin (area under the curve = 6) and paclitaxel (200 mg/m2) x 2 cycles (q21 days) were given. Objective responders received definitive radiotherapy (XRT), 70 Gy/7 weeks with concurrent weekly paclitaxel. Initially, the dose of paclitaxel was 50 mg/m2/week; because of mucosal toxicity it was reduced to 30 mg/m2/week. Patients with N2-3 disease received post-XRT neck dissection and 2 more cycles of "adjuvant" chemotherapy. In the first 22 patients, the neutropenic fever rate was 27%. Although there has been no grade IV-V toxicity from induction therapy, grade II-III toxicity resulted in an unacceptable delay in starting XRT in 14% of patients. The response rate to induction chemotherapy was 91%. Grade III mucositis occurred in all patients during concurrent chemoradiotherapy. One patient died of pneumonia during concurrent chemoradiotherapy after receiving 26 Gy and 3 doses of paclitaxel 50 mg/m2. No dose-limiting toxicity occurred in 15 patients treated with concurrent paclitaxel 30 mg/m2/week. Actuarial overall survival at 18 months is 82%; local-regional control is 86%. To date, distant metastases have not developed in any patients. This regimen has intense but acceptable acute toxicity. The maximum tolerated dosage of weekly paclitaxel during standard continuous-course XRT is confirmed to be 30 mg/m2/week. The treatment efficacy of this regimen (response rate and short-term local-regional and distant control) is encouraging. Accrual continues to obtain long-term toxicity, efficacy, and quality-of-life data. PMID- 11039516 TI - Synchronous ovarian and endometrial malignancies. AB - Synchronous ovarian primaries are infrequently found in patients with endometrial cancer. Although numerous investigators have examined the characteristics of these women, most include patients with tumors of similar histology, which may simply represent ovarian metastases. To overcome this problem, we present here patients found to have tumors of dissimilar histology. Of 499 patients with endometrial cancer undergoing primary surgery between 1980 and 1997, 18 (3.6%) were found to have endometrial and ovarian primaries of dissimilar histology. The median age was 64.2 years. Most had stage I, grades I and II, minimally invasive endometrial adenocarcinomas and stage IA mucinous or serous ovarian cystadenocarcinomas. Most ovarian tumors were either borderline or grades I and II. The 5-year actuarial disease-free (DFS) and cause-specific survivals of the entire group were 81.2% and 89.5%, respectively. Those with both stage I ovarian and endometrial primaries had a trend to a better DFS (100 versus 68.6%, p = 0.07) than did women with higher stage disease. Our data demonstrate that synchronous ovarian primaries of dissimilar histology are infrequently found in women undergoing surgery for endometrial cancer. These women seek treatment at a relatively advanced age, and have early-stage, low grade disease in both sites. Their outcome is favorable, particularly those with stage I disease in both sites. PMID- 11039517 TI - Radiation therapy in cancer patients 80 years of age and older. AB - There is a paucity of clinical data regarding radiation therapy in elderly patients. This is a retrospective study of all patients aged 80 years and older who underwent treatment with external beam irradiation at a single site. There were a total of 183 patients treated with 226 courses of therapy. The mean age was 84 years (range: 80-98 years). Fifty-eight percent of the patients were male. The treatment was deemed palliative in 51% and curative in 49%. The primary cancer diagnoses were: prostate 36, lung 28, breast 25, head and neck 23, gastrointestinal 21, hematologic 12, gynecologic 11, skin 11, genitourinary 9, unknown primary 6, central nervous system 1. The patients were able to complete the prescribed therapy in 173 of 226 courses (77%). Treatment breaks during the radiation courses were required in 81 (36%) of the courses. Radiation therapy can be safely administered to an elderly population with both curative and palliative intent with the expectation of completion in more than 80% of patients. The reasons for inability to complete therapy as prescribed are multifactorial, but careful patient selection and attention to comorbidity may optimize outcome. Further research is needed to better define these parameters. PMID- 11039518 TI - Radiation recall dermatitis induced by methotrexate in a patient with Hodgkin's disease. AB - Radiation recall dermatitis refers to an inflammatory skin reaction at a previously irradiated field subsequent to chemotherapy administration. A number of antineoplastic agents have been reported to cause this phenomenon. We observed radiation recall dermatitis in a patient with stage IV nodular sclerosing Hodgkin's disease after methotrexate therapy for acute graft-versus-host disease (GVHD) prophylaxis. The patient had previously undergone matched related bone marrow transplantation with busulfan and cyclophosphamide as a preparative regimen. Subsequently, she received cyclosporine and methotrexate for acute GVHD prophylaxis. Two areas of skin previously irradiated to 3,000 cGy developed radiation recall dermatitis after two doses of methotrexate given 2 days apart and exacerbated by the third and fourth doses. This reaction occurred 34 days after the last dose of radiation therapy (RT). We believe this is the first case of radiation recall dermatitis after methotrexate therapy. Given the increased use of methotrexate in several neoadjuvant and adjuvant protocols in association with RT, its potential to produce radiation recall reactions should be considered. PMID- 11039520 TI - Learning from patients. PMID- 11039521 TI - High stereochemical diversity and applications for the synthesis of marine natural products: a library of carbohydrate mimics and polyketide segments AB - We have developed a powerful concept for the rapid assembly of a series of twenty four homochiral building blocks from simple racemic trans-2,4-dimethyl-8 oxabicyclo[3.2.1]oct-6-en-3-one. The series comprises eight stereochemical pentades of anomeric [3.3.1]lactone acetals, eight stereochemical tetrades of anomeric carbohydrate mimics, and eight stereotetrades of acyclic polypropionate units. The utility of these enantiopure materials (average 94% ee) in natural product synthesis is demonstrated and shown to complement the popular aldol method. PMID- 11039519 TI - Phase II evaluation of continuous-infusion 5-fluorouracil, leucovorin, mitomycin C, and oral dipyridamole in advanced measurable pancreatic cancer: a North Central Cancer Treatment Group Trial. AB - At present there remains a need for more effective systemic therapy in advanced pancreatic cancer. Some studies have suggested that infusional chemotherapy schedules and biomodulation of 5-fluorouracil (5-FU) may improve the therapeutic outcome in advanced colon cancer. One such regimen that uses continuous infusion 5-FU, weekly leucovorin, daily dipyridamole, and intermittent mitomycin-C has activity in both colon and unresectable pancreatic carcinoma. The intent of this trial was to test the effectiveness of this four-drug regimen in advanced pancreatic cancer. Patients received 5-FU 200 mg/m2 daily by continuous infusion, leucovorin 30 mg/m2 IV weekly, mitomycin-C 10 mg/m2 day 1, and dipyridamole 75 mg orally four times daily for 5 weeks. After a 1-week break, treatment cycles were repeated every 6 weeks. Eligibility included biopsy-proven advanced measurable pancreatic cancer, Eastern Cooperative Oncology Group performance status 0 and 2, and no prior systemic chemotherapy. Of 46 evaluable patients, 9 partial responses and 1 complete tumor response were seen, for an overall response rate of 22% (95% confidence interval 11-36%). The median survival in the group of 50 patients registered to this trial was 4.6 months, with a range of 0.33 to 40.2 months. Toxicity was manageable, with the most common toxicities (> or =grade III National Cancer Institute Common Toxicity Criteria) being anorexia (13%), stomatitis (17%), and hand-foot syndrome (13%). Of note, little severe hematologic toxicity and no significant headaches were reported. Although some patients did respond, the therapeutic results are not encouraging enough to take this regimen to phase III testing. PMID- 11039522 TI - Maximizing synthetic efficiency: multi-component transformations lead the way AB - With the emergence of high-throughput screening in the pharmaceutical industry in the early 1990's, organic chemists were faced with a new challenge: how to prepare large collections of molecules (the libraries) to "feed" the high throughput screen? The unique exploratory power of some reactions (such as the 40 year-old Ugi four-component condensation) was soon recognized to be extremely valuable to produce libraries in a time- and cost-effective manner. Over the last five years, industrial and academic researchers have made these powerful transformations into one of the most efficient and cost-effective tools for combinatorial and parallel synthesis. PMID- 11039523 TI - Optically active dioxatetraazamacrocycles: chemoenzymatic syntheses and applications in chiral anion recognition AB - Two new C2 and D2 symmetrical dioxatetraaza 18-membered macrocycles [(R,R)-1 and (S,S,S,S)-2] are efficiently synthesized in enantiomerically pure forms by a chemoenzymatic method starting from (+/-)-trans-cyclohexane-1,2-diamine. The protonation constants and the binding constants with different chiral dicarboxylates are determined in aqueous solution by means of pH-metric titrations. The triprotonated form of (S,S,S,S)-2 shows moderate enantioselectivity with malate and tartrate anions (deltadeltaG=0.62 and 0.66 kcal mol(-1), respectively), being the strongest binding observed in both cases with the L enantiomer. Good enantiomeric discrimination is obtained with tetraprotonated (R,R)-1 and N-acetyl aspartate, the complex with the D-enantiomer being 0.92 kcalmol(-1) more stable than its diastereomeric counterpart. Despite the lack of enantioselectivity of tri- and tetraprotonated (R,R)-1 for the tartrate anion, a very good diastereopreference for meso-tartrate is found. All these experimental results allow us to propose a model for the host-guest structure based on coulombic interactions and hydrogen bonds. PMID- 11039524 TI - Temperature rule for the speed of sound in water: a chemical kinetics model AB - Water forms three-dimensional polymeric structures due to the influence of hydrogen bonds and is fundamentally different from other substances. One of the simplest ways to analyze the structure of water in any system, such as hydration, is to measure the degree of compressibility, which can be determined from the speed of sound, by making use of the physical laws established by Newton and later perfected by Laplace. Although the speed of sound is strongly dependent on the temperature of a liquid, Laplace's equation does not refer to temperature in any of its terms. It is necessary, therefore, to determine the degree of temperature dependency. However, only approximate expressions of a fifth-order polynomial have been reported so far in the literature. In this paper, a universal method for describing the speed of sound from the perspective of physicochemical reaction kinetics is presented. It is shown that the speed of sound U [ms(-1)] changes with temperature T [K] according to a thermodynamically derived formula given as U= exp(-A/T-BlnT+C) and that the motion and propagation phenomena of sound energy can also be regarded as chemical reactions. PMID- 11039525 TI - Conformation and stability of the hydrogen-bonded complex 6-oxabicyclo AB - The hydrogen-bonded complex between 6-oxabicyclo[3.1.0]hexane and hydrogen chloride was investigated by microwave spectroscopy in a supersonic jet. A dual flow pulsed valve was used to preclude chemical reaction between the monomers. Only the equatorial conformer was observed and the spectra of three isotopomers, (C5H8O, H35Cl), (C5H8O, H37Cl) and (C5H8O, D35Cl), were measured. The derived structure of the complex has Cs symmetry with the hydrogen chloride pointing to the domain of the equatorial lone pair of electrons at the oxygen atom. The three atoms involved in the hydrogen bond adopt a bent arrangement with a O...H distance of 1.77(4) A, a (O...H-Cl) angle of 115(4)degrees, and a deviation of 15.4(14)degrees of the hydrogen bond from collinearity. In agreement with the experimental results, ab initio calculations predict the equatorial form to be the most stable one. PMID- 11039526 TI - Giant vesicles from 72-membered macrocyclic archaeal phospholipid analogues: initiation of vesicle formation by molecular recognition between membrane components AB - Stereochemically pure archeal acyclic bola-amphiphilic diphosphates 4 and 5, with the basic structure of the phospholipids found in Sulfolobus, have been synthesized for the first time. The self-assembly properties have been compared with those of the nearly identical 72-membered macrocyclic tetraether phosphates 3a and 3b, analogues of the major phospholipid components of Sulfolobus, Thermoplasma, and methanogenic Archea, which were also synthesized. Phase contrast and fluorescence microscopies have shown that the dipolar lipids 1 and 2 spontaneously formed vesicles. Whereas the macrocyclic dipolar phosphates 3 spontaneously formed vesicles (phase contrast and fluorescence microscopies), the bolaform phosphate 4 gave only a lamellar structure (synchrotron diffraction pattern: repeat distance of about 4.25 nm but with only a few layers). However, upon addition of the unphosphorylated precursors phytanol, phytol, or geranylgeraniol to the acyclic lipids 4 and 5, giant vesicles were rapidly formed. Addition of n-hexadecanol or cholesterol did not lead to vesicle formation. Therefore it was concluded that this vesicle formation occurs only when the added molecule is closely compatible with the constituents of the lipid layer and can be inserted into the double layer. A slight mismatch (cholesterol or n-hexadecanol/polyprenyl chains) is therefore enough to block the insertion process presumably required for vesicle formation. PMID- 11039527 TI - Highly enantiomerically enriched alpha-haloalkyl Grignard reagents AB - alpha-Chloro- and alpha-bromoalkyl Grignard reagents 11 and 30 with > 97% ee (enantiomeric excess) were generated by a sulfoxide/magnesium exchange reaction from the enantiomerically and diastereomerically pure sulfoxides 25 and 27. The resulting alpha-haloalkyl Grignard reagents are configurationally stable at -78 degrees C. Racemization sets in at or above -60 degrees C, especially when the solution contains bromide ions. In the absence of halide ions, the configurational stability extends up to -20 degrees C, when chemical decomposition commences. PMID- 11039528 TI - Synthesis and molecular geometry of an achiral 30-crown-12 polyacetal from alpha cyclodextrin AB - Periodate oxidation of alpha-cyclodextrin followed by borohydride reduction readily provided an octadeca-hydroxymethyl-substituted 30-crown-12 polyacetal 1, its 30-membered macrocycle being composed of six meso-butanetetrol/glycolaldehyde acetal units, which is, consequently, optically inactive. Its solid-state molecular geometry emerged from the X-ray structural analysis of the well crystallizing octadeca-acetate 2, which revealed the undulated macrocycle to be molded into three loops with a unique order of succession of the -CHR-CHR-O-CHR-O units: alternating gauche- and anti-conformations of the meso-butanetetrol portions and consecutive disposition of the glycolaldehyde-acetoxymethyl groups above and below the mean-plane of the macrocycle. In solution, however, as evidenced by 1H- and 13C-NMR spectra, the macrocycle is highly flexible at ambient and higher temperatures, its mobility becoming distinctly restricted only below -20 degrees C. PMID- 11039529 TI - Oxidative addition of allylic carbonates to palladium(0) complexes: reversibility and isomerization AB - The oxidative addition of a cyclic allylic carbonate to the palladium(0) complex generated from a [Pd(dba)2]+2 PPh3 mixture affords a cationic pi allylpalladium(II) complex with the alkyl carbonate as the counter-anion. This reaction is reversible and proceeds with isomerization of the allylic carbonate at the allylic position. The equilibrium constant has been determined in DMF. The influence of the precursor of the palladium(0) is discussed. PMID- 11039530 TI - Syntheses and 1H-, 13C- and 15N-NMR spectra of ethynyl isocyanide, H-C triple bond C-N triple bond C, D-C triple bond C-N triple bond C and prop-1-ynyl isocyanide, H3C-C triple bond C-N triple bond C, D3C-C triple bond C-N triple bond C: high resolution infrared specturm of prop-1-ynyl isocyanide AB - Ethynyl isocyanide, H-C triple bond C-N triple bond C (1a), deuteroethynyl isocyanide, D-C triple bond C-N triple bond C (1b), prop-1-ynyl isocyanide, H3C-C triple bond C-N triple bond C (1c), and trideuteroprop-1-ynyl isocyanide, D3C-C triple bond C-N triple bond C (1d) are synthesized by flash vacuum pyrolysis of suitable organometallic precursor molecules (CO)5Cr(CN-CCl triple bond CClH) (5a), (CO)5Cr(CN-CCI=CClD) (5b), (CO)5Cr(CN-CCl=CCl-CH3) (5c) and (CO)5Cr(CN CCI=CCl-CD3) (5d), respectively. Compounds 5a-d are formed in two steps by radical alkylation of tetraethyl-ammonium pentacarbonyl(cyano)chromate, NEt4[Cr(CO)5(CN)] (2) by 1,1,2,2,-tetrachloroethane (3a), 1,1,2,2-tetrachloro-1,2 dideuteroethane (3b), 1,1,2,2,-tetrachloropropane (3c), and 1,1,2,2-tetrachloro- 1,3,3,3-tetradeutero-propane (3d) yielding [(CO)5Cr(CN-CCl2-CCl2-H)] (4a), [(CO)5Cr(CN-CCl2-CCl2D)] (4b), [(CO)5Cr(CN-CCl2-CCl2-CH3)] (4c), and [(CO)5Cr(CN CCl2-CCl2-CD3)] (4d). Dehalogenation of 4a-d using zinc in diethylether/acetic acid gives 5a-d, respectively. A multinuclear NMR study revealed the 1H-, 13C- and 15N-NMR data of 1a and 1c. Molecular spectroscopic data of 1c were determined by high resolution infrared spectroscopy. The by-products of the pyrolysis are the E and Z isomers of the halogenated ethenyl isocyanides H(Cl)C=CCl-NC (6a) and H3C(Cl)C=CCl-NC (6c) which have been characterized by IR, MS and NMR spectroscopy. PMID- 11039531 TI - Phenylalanine ammonia-lyase: the use of its broad substrate specificity for mechanistic investigations and biocatalysis--synthesis of L-arylalanines. AB - Several fluoro- and chlorophenylalanines were found to be good substrates of phenylalanine ammonia-lyase (PAL/EC 4.3.1.5) from parsley. The enantiomerically pure L-amino acids were obtained in good yields by reaction of the corresponding cinnamic acids with 5M ammonia solution (buffered to pH 10) in the presence of PAL. The kinetic constants for nine different fluoro- and chlorophenylalanines do not provide a rigorous proof for but are consistent with the previously proposed mechanism comprising an electrophilic attack of the methylidene-imidazolone cofactor of PAL at the aromatic nucleus as a first chemical step. In the resulting Friedel-Crafts-type sigma complex the beta-protons are activated for abstraction and consequently the pro-S is abstracted by an enzymic base. Results from semi-empirical calculations combined with a proposed partial active site model showed a correlation between the experimental kinetic constants and the change in polarization of the pro-S Cbeta-H bond and heat of formation of the sigma complexes, thus making the electrophilic attack at the neutral aromatic ring plausible. Furthermore, while 5-pyrimidinylalanine was found to be a moderately good substrate of PAL, 2-pyrimidinylalanine was an inhibitor. PMID- 11039532 TI - Steric inhibition of resonance: a revision and quantitative estimation on the basis of aromatic carboxylic acids AB - The classical concepts of steric inhibition of resonance (SIR) and primary steric effect (van der Waals interactions) were revised with the aid of methyl substituted benzoic acids. The quantum chemical model was based on the energies of various conformations, calculated at the RHF/6-31 +G(d,p) and B3LYP/6-311 + G(3df,2pd)//RHF/6-31 +G(d,p) levels. The molecule of 2-methylbenzoic acid is planar: no SIR is possible, and van der Waals interaction is practically equal in the acid molecule and in its anion. Therefore, the increased strength of this acid is not due to any steric effect but can be described in terms of electrostatic interaction pole/induced dipole, which lowers the energy of the anion. The molecule of 2,6-dimethylbenzoic acid is nonplanar. SIR is significant in the acid molecule but equal or even greater in the anion. The higher acidity cannot be connected with SIR and can be explained also by electrostatic interaction. In 2,3,5,6-tetramethylbenzoic acid, SIR is greater and may be responsible for one third of the acid-strengthening effect. The concept of SIR is to be applied with caution; even when the nonplanar conformation is proven, SIR need not be responsible for any observable quantity, and particularly not for the acidic and basic properties. PMID- 11039533 TI - Novel artificial receptors for alkylammonium ions with remarkable selectivity and affinity AB - The benzene-based tripodal tris(oxazolines) have been developed as the most selective and strong receptors toward linear alkylammonium ions reported to date. Among six tris(oxazolines) based on 2,4,6-trimethylbenzene framework, the phenylglycinol-derived receptor 4 exhibits the largest association constant toward nBuNH3+ (logK(ass) = 6.65 +/- 0.02), while a similar value toward tBuNH3+, (logK(ass) = 3.80 +/- 0.01) compared with others, which corresponds to the selectivity ratio of nBuNH3+/tBuNH3+ as high as approximately equals 700. The tris(oxazoline) 6 that has bare oxazoline ring exhibits still a large association constant toward sterically hindered tBuNH3+ (logK(ass) = 5.26 +/- 0.02). Both receptors 4 and 6 extract beta-phenethylammonium ion from water into chloroform almost completely. When the benzene frame is changed from 2,4,6-trimethylbenzene to 2,4,6-triethylbenzene, dramatic changes in the affinity as well as in the selectivity are observed. The association constant observed by tris(oxazoline) 8 toward nBuNH3+ approaches 10(8)M(-1) and the selectivity ratio of nBuNH3+/tBuNH3+ is increased to 2,700. This selectivity is even more enhanced to 4,000 with tris(oxazoline) 9. The enhanced binding affinity and high selectivity observed with receptors 4 and related derivatives 7-9 compared with others can be explained by an optimized steric and electronic environment provided by the phenyl substituents, which has been unambiguously demonstrated by X-ray crystallographic and 1H NMR spectroscopic studies on the host-guest complexes. The new receptor system has several unique features such as ready availability, structural simplicity, and in particular, versatility in derivatization. By virtue of these advantages, it can be readily tailored as selective receptors toward biologically important amines. PMID- 11039534 TI - The kinetics of substitution reactions of mixed platinum(II)-bis(nucleoside) complexes in aqueous solution in the presence of thiourea; X-ray crystal structure determination of cis- AB - In aqueous solution, bis(nucleoside) complexes formed by the reaction of cis [Pt(NH3)2(H2O)2]2+ with an excess of either adenosine (ado) or a mixture of adenosine and guanosine (guo) undergo a slow N7--> N1 linkage isomerisation in the adenine moiety. The isomerisation probably involves the breaking and reformation of Pt-nucleoside bonds, thus favouring the more stable N1 binding mode of the adenine base. Dynamic processes due to the presence of adenosine in the platinum coordination sphere are slow on the NMR time scale. The N7 binding mode of PtII in cis-[Pt(NH3)2(ado-N7)2](ClO4)2. 3.5H2O was confirmed by X-ray crystal structure analysis. In both of the crystallographically independent cations, the PtII coordination sphere is almost ideally square planar, with typical Pt-N bond lengths and angles. The most significant difference between the two cations lies in the sugar conformation of the coordinated nucleosides. In one cation, both have an anti (-ap) conformation, whilst in the other cation one has an anti (-ap) conformation and the other a syn (+sc) conformation stabilised by a relatively strong H-bond. Substitution of the nucleoside(s) by thiourea follows an associative mechanism with only a negligible contribution by the solvent path. For symmetric complexes, the order of lability of different binding modes is ado N1 3)-4,6-O-benzylidene-2-a cetamido-2-deoxy-alpha-D galactopyranoside with a novel sialyl donor. A tetrasaccharide, pentasaccharide, and hexasaccharide were constructed in predictable and controlled manner with high regio- and stereoselectivity after the successful preparation and employment of a disaccharide donor, trisaccharide donor, disaccharide acceptor, and trisaccharide acceptor building blocks. Finally, a mild oxidative cleaving method was adopted for the selective removal of 2-naphthylmethyl (NAP) in the presence of benzyl groups. PMID- 11039539 TI - Conformational transition of acidic peptides exposed to minerals in suspension AB - Mineral surfaces probably participated in the chemical processes which led to life in the primitive oceans. The ordered conformations of simple acidic peptides exposed to insoluble minerals are described. Alternating poly(Glu-Leu) adopts a random coil conformation in water due to charge repulsion. The polypeptide extracts cations from insoluble crystalline CdS or molybdenum and adopts an ordered conformation. CdS leads to the formation of beta-sheets whereas molybdenum leads to alpha-helices. Peptides with at least 10-amino acids are necessary to exhibit a significative adsorption onto the surface. Under the same conditions, montmorillonite adsorbs the polypeptide but does not induce any conformational change. PMID- 11039540 TI - Dye-loaded zeolite L sandwiches as artificial antenna systems for light transport AB - The synthesis and characterization of dye loaded zeolite L sandwiches acting as artificial antenna systems for light harvesting and transport is reported. A set of experimental tools for the preparation of neutral dye-zeolite L materials ranging from low to maximum packing densities has been developed. The role of co adsorbed water and the distribution of molecules between the inner and the outer surface were found to be the determining parameters. p-Terphenyl (pTP) turned out to be very suitable for studying these and other relevant parameters of neutral dye-zeolite L materials. We observed that pTP located in the channels of zeolite L can reversibly be displaced by water. This can be used when synthesizing such materials. We also observed that all-trans-1,6-diphenyl-1,3,5-hexatriene (DPH) which is very photolabile in solution is stable after insertion into zeolite L. By combining our extensive knowledge of these systems with ion-exchange procedures developed in an earlier study, we have realized the first bi directional three-dye antenna. In this material the near UV absorbing compounds DPH or 1,2-bis-(5-methyl-benzoxazol-2-yl)-ethene (MBOXE) are located in the middle part of zeolite L nanocrystals followed on both sides by pyronine (Py) and then by oxonine (Ox) as acceptors. Fluorescence of the oxonine located at both ends of the cylindrical zeolite L crystals was observed upon excitation of the near UV absorber in the middle section at 353 nm, where neither oxonine nor pyronine absorb a significant amount of the excitation light. PMID- 11039541 TI - Association between changes in eating and drinking behaviors and respiratory tract disease in newly arrived calves at a feedlot. AB - OBJECTIVE: To investigate eating and drinking behaviors and their association with bovine respiratory disease complex (BRDC) and to evaluate methods of diagnosing BRDC. ANIMALS: 170 newly arrived calves at a feedlot. PROCEDURE: Eating and drinking behaviors of calves were recorded at a feedlot. Calves with clinical signs of BRDC were removed from their pen and classified retrospectively as sick or not sick on the basis of results of physical and hematologic examinations. Pulmonary lesions of all calves were assessed at slaughter. RESULTS: Calves that were sick had significantly greater frequency and duration of drinking 4 to 5 days after arrival than calves that were not sick. Sick calves had significantly lower frequency and duration of eating and drinking 11 to 27 days after arrival but had significantly greater frequency of eating 28 to 57 days after arrival than calves that were not sick. Calves at slaughter that had a higher percentage of lung tissue with pneumonic lesions had significantly lower frequency and duration of eating 11 to 27 days after arrival but had significantly higher frequency and duration of eating 28 to 57 days after arrival. Agreement for calves being sick and having severe pulmonary lesions at slaughter was adequate. Agreement for calves being removed and having pulmonary lesions at slaughter was low. CONCLUSIONS AND CLINICAL RELEVANCE: Eating and drinking behaviors were associated with signs of BRDC, but there was not an obvious predictive association between signs of BRDC in calves and eating and drinking behaviors. Fair to poor agreement was observed between antemortem and postmortem disease classification. PMID- 11039542 TI - Lesions and effects of location for administration of clostridial bacterin-toxoid vaccines on growth performance and eating and drinking behaviors in newly arrived calves at a feedlot. AB - OBJECTIVES: To determine the effect of location for administration of clostridial vaccines on behavior, growth performance, and health of calves at a feedlot, the relative risk of calves developing an injection-site reaction or being misdiagnosed as having bovine respiratory disease complex (BRDC), and the percentage of subcutaneous injection-site reactions that were detectable on carcasses after the hides were removed. ANIMAL: 170 newly arrived calves at a feedlot. PROCEDURE: Eating and drinking behaviors of calves during the initial 57 days after arrival were observed at a commercial feedlot, using an electronic monitoring system. Calves were assigned randomly to receive a clostridial vaccine (base of ear or neck). Data on reactions at the injection site were collected. RESULTS: Mean daily gain (MDG) for the initial 57 days did not differ significantly between treatments. Risk of being misdiagnosed as having BRDC was not associated with location for administration of vaccine. Calves vaccinated in the base of the ear were at higher risk of having an injection-site reaction at day 57 or at slaughter. Eighty-nine percent (95% confidence interval, 52 to 100%) of injection-site reactions in the neck could not be located on the carcasses after hides were removed. Calves vaccinated in the neck drank significantly fewer times per day during the first 57 days than calves vaccinated in the base of the ear. CONCLUSIONS AND CLINICAL RELEVANCE: Location for administration of a clostridial vaccine did not significantly affect health, growth performance, or eating behavior. Most subcutaneous injection-site reactions were not detectable after the hide was removed. PMID- 11039543 TI - Nitric oxide generation in a rat model of acute portal hypertension. AB - OBJECTIVE: To document blood nitric oxide concentrations in the portal vein and systemic circulation in a rat model of acute portal hypertension and compare values with a control group and a sham surgical group. ANIMALS: 30 rats; 10 controls (group 1), 10 sham surgical (group 2), and 10 rats with surgically induced acute portal hypertension (group 3). PROCEDURE: Following induction of anesthesia, catheters were placed surgically in the carotid artery, jugular, and portal veins of group 2 and 3 rats and in the carotid artery and jugular vein of group 1 rats. Baseline heart and respiratory rates, rectal temperature, and vascular pressure measurements were obtained, and blood was drawn from all catheters for baseline nitric oxide (NO) concentrations. Acute portal hypertension was induced in the group 3 rats by tying a partially occluding suture around the portal vein and a 22-gauge catheter. The catheter was then removed, resulting in a repeatable degree of portal vein impingement. After catheter placement, all variables were remeasured at 15-minute intervals for 3 hours. RESULTS: Blood nitric oxide concentrations were greater in all vessels tested in group 3 than in group 2 rats. CONCLUSIONS AND CLINICAL RELEVANCE: Acute portal hypertension in this experimental model results in increased concentrations of NO in the systemic and portal circulation. On the basis of information in the rat, it is possible that increased NO concentrations may develop in dogs following surgical treatment of congenital portosystemic shunts if acute life-threatening portal hypertension develops. Increased NO concentrations may contribute to the shock syndrome that develops in these dogs. PMID- 11039544 TI - Evaluation of substance P as a neurotransmitter in equine jejunum. AB - OBJECTIVE: To determine whether substance P (SP) functions as a neurotransmitter in equine jejunum. SAMPLE POPULATION: Samples of jejunum obtained from horses that did not have lesions in the gastrointestinal tract. PROCEDURE: Jejunal smooth muscle strips, oriented in the plane of the circular or longitudinal muscle, were suspended isometrically in muscle baths. Neurotransmitter release was induced by electrical field stimulation (EFS) delivered at 2 intensities (30 and 70 V) and various frequencies on muscle strips that were maintained at low tension or were under contraction. A neurokinin-1 receptor blocker (CP-96,345) was added to baths prior to EFS to interrupt SP neurotransmission. Additionally, direct effects of SP on muscle strips were evaluated, and SP-like immunoreactivity was localized in intestinal tissues, using indirect immunofluorescence testing. RESULTS: Substance P contracted circularly and longitudinally oriented muscle strips. Prior treatment with CP-96,345 altered muscle responses to SP and EFS, suggesting that SP was released from depolarized myenteric neurons. Depending on orientation of muscle strips and stimulation variables used, CP-96,345 increased or decreased the contractile response to EFS. Substance P-like immunoreactivity was detected in the myenteric plexus and circular muscle layers. CONCLUSIONS AND CLINICAL RELEVANCE: Substance P appears to function as a neurotransmitter in equine jejunum. It apparently modulates smooth muscle contractility, depending on preexisting conditions. Effects of SP may be altered in some forms of intestinal dysfunction. Altering SP neurotransmission in the jejunum may provide a therapeutic option for motility disorders of horses that are unresponsive to adrenergic and cholinergic drugs. PMID- 11039545 TI - Concentrations of gentamicin in serum and bronchial lavage fluid after intravenous and aerosol administration of gentamicin to horses. AB - OBJECTIVE: To compare concentrations of gentamicin in serum and bronchial lavage fluid after IV and aerosol administration of gentamicin to horses. ANIMALS: 9 healthy adult horses. PROCEDURE: Gentamicin was administered by aerosolization (20 ml of gentamicin solution [50 mg/ml]) and IV injection (6.6 mg of gentamicin/kg of body weight) to each horse, with a minimum of 2 weeks between treatments. Samples of pulmonary epithelial lining fluid were collected by small volume (30 ml) bronchial lavage 0.5, 4, 8, and 24 hours after gentamicin administration. Serum samples were obtained at the same times. All samples were analyzed for gentamicin concentration, and cytologic examinations were performed on aliquots of bronchial lavage fluid collected at 0.5, 8, and 24 hours. RESULTS: Gentamicin concentrations in bronchial lavage fluid were significantly greater 0.5, 4, and 8 hours after aerosol administration, whereas serum concentrations were significantly less at all times after aerosol administration, compared with IV administration. Neutrophil counts in bronchial lavage fluid increased from 0.5 to 24 hours, regardless of route of gentamicin administration. CONCLUSIONS AND CLINICAL RELEVANCE: Aerosol administration of gentamicin to healthy horses resulted in gentamicin concentrations in bronchial fluid that were significantly greater than those obtained after IV administration. A mild inflammatory cell response was associated with aerosol delivery of gentamicin and repeated bronchial lavage. Aerosol administration of gentamicin may have clinical use in the treatment of bacterial bronchopneumonia in horses. PMID- 11039546 TI - Effects of intravenous administration of formaldehyde on platelet and coagulation variables in healthy horses. AB - OBJECTIVES: To assess safety and determine effects of IV administration of formaldehyde on hemostatic variables in healthy horses. ANIMALS: 7 healthy adult horses. PROCEDURE: Clinical signs and results of CBC, serum biochemical analyses, and coagulation testing including template bleeding time (TBT) and activated clotting time (ACT) were compared in horses given a dose of 0.37% formaldehyde or lactated Ringer's solution (LRS), IV, in a 2-way crossover design. In a subsequent experiment, horses received an infusion of 0.74% formaldehyde or LRS. In another experiment, horses were treated with aspirin to impair platelet responses prior to infusion of formaldehyde or LRS. RESULTS: Significant differences were not detected in any variable measured between horses when given formaldehyde or any other treatment. Infusion of higher doses of formaldehyde resulted in adverse effects including muscle fasciculations, tachycardia, tachypnea, serous ocular and nasal discharge, agitation, and restlessness. CONCLUSIONS AND CLINICAL RELEVANCE: Intravenous infusion of formaldehyde at doses that do not induce adverse reactions did not have a detectable effect on measured hemostatic variables in healthy horses. PMID- 11039547 TI - Evaluation of changes in hematologic and clinical biochemical values after exposure to petroleum products in mink (Mustela vison) as a model for assessment of sea otters (Enhydra lutris). AB - OBJECTIVE: To determine the effects of petroleum exposure on hematologic and clinical biochemical results of mink and to identify variables that may be useful for making management decisions involving sea otters (Enhydra lutris) that have been exposed to oil in their environment. ANIMALS: 122 American mink (Mustela vison). PROCEDURES: Mink were exposed once to a slick of oil (Alaskan North Slope crude oil or bunker C fuel oil) on seawater or via low-level contamination of their daily rations. RESULTS: In the acute phase of exposure, petroleum directly affected RBC, WBC, neutrophil, and lymphocyte counts, fibrinogen, sodium, calcium, creatinine, total protein, and cholesterol concentrations, and alanine transaminase, creatine kinase, alkaline phosphatase, and gamma glutamyltransferase activities. Aspartate transaminase, alkaline phosphatase, gamma-glutamyltransferase, and lactate dehydrogenase activities and cholesterol concentration also varied as a result of chronic low-level contamination of feed. CONCLUSIONS AND CLINICAL RELEVANCE: Our results are in agreement with reports that attribute increased alanine transaminase and alkaline phosphatase activities and decreased total protein concentration to petroleum exposure in sea otters during an oil spill. Sodium, calcium, creatinine, cholesterol, and lactate dehydrogenase may be valuable variables to assess for guidance during initial treatment of sea otters exposed to oil spills as well as for predicting which petroleum-exposed sea otters will reproduce following an oil spill. Measurement of these variables should aid wildlife professionals in making decisions regarding treatment of sea otters after oil spills. PMID- 11039548 TI - Pharmacokinetics of ceftazidime in dogs following subcutaneous administration and continuous infusion and the association with in vitro susceptibility of Pseudomonas aeruginosa. AB - OBJECTIVE: To determine the pharmacokinetics of ceftazidime following subcutaneous administration and continuous IV infusion to healthy dogs and to determine the minimum inhibitory concentration (MIC) of ceftazidime for clinical isolates of Pseudomonas aeruginosa. ANIMALS: 10 healthy adult dogs. PROCEDURE: MIC of ceftazidime for 101 clinical isolates of P aeruginosa was determined in vitro. Serum concentrations of ceftazidime were determined following subcutaneous administration of ceftazidime (30 mg/kg of body weight) to 5 dogs and continuous IV infusion of ceftazidime (loading dose, 4.4 mg/kg; infusion rate, 4.1 mg/kg/h) for 36 hours to 5 dogs. RESULTS: The MIC of ceftazidime for P aeruginosa was < or = 8 microg/ml; all isolates were considered susceptible. Following SC administration of ceftazidime, mean beta disappearance half-life was 0.8 hours, and mean serum ceftazidime concentration exceeded the MIC for P aeruginosa for only 4.3 hours. Two dogs had gastrointestinal tract effects. Mean serum ceftazidime concentration exceeded 16 microg/ml during continuous IV infusion. None of the dogs developed adverse effects. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of ceftazidime subcutaneously (30 mg/kg, q 4 h) or as a constant IV infusion (loading dose, 4.4 mg/kg; rate, 4.1 mg/kg/h) would maintain serum ceftazidime concentrations above the MIC determined for 101 clinical isolates of P aeruginosa. Use of these dosages may be appropriate for treatment of dogs with infections caused by P aeruginosa. PMID- 11039549 TI - Evaluation of the ability of orally administered aspirin to mitigate effects of 3 methylindole in feedlot cattle. AB - OBJECTIVE: To evaluate the ability of orally administered aspirin to mitigate 3 methylindole (3MI)-induced respiratory tract disease and reduced rate of gain in feedlot cattle. ANIMALS: 244 beef cattle. PROCEDURE: In a masked, randomized, controlled field trial, calves were untreated (controls) or received a single orally administered dose of aspirin (31.2 g) on entry into a feedlot. Serum 3MI concentrations were measured on days 0, 3, and 6. Rumen 3MI concentration was measured on day 3. Cattle were observed daily for clinical signs of respiratory tract disease. Lungs were evaluated at slaughter for gross pulmonary lesions. RESULTS: Mean daily gain (MDG) in cattle treated with aspirin, compared with control cattle, was 0.06 kg greater in the backgrounding unit and 0.03 kg greater for the overall feeding period. Neither serum nor rumen 3MI concentrations appeared to modify this effect. Cattle treated with aspirin were more likely to be treated for respiratory tract disease. Mortality rate, gross pulmonary lesions, and serum and rumen 3MI concentrations were similar between groups. Increased rumen 3MI concentration was associated with a small difference in risk of lung fibrosis. CONCLUSIONS AND CLINICAL RELEVANCE: Cattle given a single orally administered dose of aspirin on feedlot entry had higher MDG in the backgrounding unit and for the overall feeding period, but this finding could not be attributed to mitigation of effects of 3MI. This may have been influenced by low peak 3MI production and slow rates of gain. PMID- 11039550 TI - Dual-energy X-ray absorptiometry measurement of bone-mineral density in the distal aspect of the limbs in racing Greyhounds. AB - OBJECTIVE: To determine those bones in the distal aspect of the limbs of Greyhounds with fatigue fractures that have the greatest left-to-right differences in bone-mineral density (BMD). SAMPLE POPULATION: Limbs obtained from 20 Greyhounds. PROCEDURE: Dual-energy x-ray absorptiometry (DXA) of the distal aspect of each limb and isolated bones from 10 dogs with a fracture of the central tarsal bone (CTB) of the right pelvic limb was performed. High-resolution scanning was performed on excised bones, and BMD measurements of CTB also were obtained from limbs of dogs without a CTB fracture. RESULTS: The BMD of the accessory carpal bone and calcaneus was not significantly different from the BMD of those bones in the contralateral limb. Although BMD of the CTB of the entire right pelvic limb and isolated bones from dogs with a CTB fracture was lower, compared with values for the entire left pelvic limb, values for isolated CTB from dogs without a CTB fracture were not significantly different. Metacarpal or metatarsal and thoracic or pelvic limb significantly affected BMD for measurements of the entire limb and isolated bones. Left-to-right differences in BMD were greatest for metacarpal 5. CONCLUSIONS AND CLINICAL RELEVANCE: Asymmetric adaptive remodeling of metacarpal 5 can be detected by DXA. The potentially confounding effects of CTB fracture and unknown racing history made it difficult to interpret BMD changes in the CTB of these specimens. Densitometry could be developed as an in vivo assessment for risk of fractures in racing Greyhounds. PMID- 11039551 TI - Effects of 0.005% latanoprost solution on intraocular pressure in healthy dogs and cats. AB - OBJECTIVE: To evaluate effects of daily topical ocular administration of latanoprost solution on intraocular pressure (IOP) in healthy cats and dogs. ANIMALS: 9 domestic shorthair cats and 14 dogs. PROCEDURE: Latanoprost solution (0.005%) was administered topically to 1 eye (treated) and vehicle to the other eye (control) of all animals once daily in the morning for 8 days. Intraocular pressure was measured twice daily for the 5 days preceding treatment, and IOP, pupillary diameter, conjunctival hyperemia, and blepharospasm were measured 0, 1, 6, and 12 hours after the first 4 treatments and 0 and 12 hours after the final 4 treatments. Measurements continued twice a day for 5 days after treatment was discontinued. Aqueous flare was measured once daily during and for 5 days after the treatment period. RESULTS: Intraocular pressure and pupillary diameter were significantly decreased in the treated eye of dogs, compared with the control eye. Mild conjunctival hyperemia was also detected, but severity did not differ significantly between eyes. Blepharospasm and aqueous flare were not detected in either eye. Intraocular pressure in cats was not significantly affected by treatment with latanoprost. However, pupillary diameter was significantly decreased in the treated eye, compared with the control eye. Conjunctival hyperemia, aqueous flare, and blepharospasm were not detected in either eye. CONCLUSIONS AND CLINICAL RELEVANCE: Once-daily topical ocular administration of latanoprost solution (0.005%) reduced IOP in healthy dogs without inducing adverse effects but did not affect IOP in healthy cats. Latanoprost may be useful for treating glaucoma in dogs. PMID- 11039553 TI - Risk factors for outbreaks of disease attributable to white sturgeon iridovirus and white sturgeon herpesvirus-2 at a commercial sturgeon farm. AB - OBJECTIVE: To determine management, fish, and environmental risk factors for increased mortality and an increased proportion of runts for white sturgeon exposed to white sturgeon iridovirus (WSIV) and white sturgeon herpesvirus-2 (WSHV-2). ANIMALS: White sturgeon in 57 tanks at 1 farm and observations made for fish at another farm. PROCEDURE: A prospective cohort study was conducted. Data on mortality, proportion of runts, and potential risk factors were collected. Five fish from each tank were examined for WSIV and WSHV-2 via inoculation of susceptible cell lines and microscopic examination of stained tissue sections. An ANCOVA was used to evaluate effects of risk factors on mortality and proportion of runts. RESULTS: Major determinants of number of dead fish (natural logarithm [In]-transformed) were spawn, source (90% confidence interval [CI] for regression coefficient, 0.62 to 2.21), and stocking density (90% CI, 0.003 to 0.03). Main predictors of proportion of runts (In-transformed) were spawn, mortality incidence density (90% CI, 0.004 to 0.03), age (90% CI, -0.012 to -0.004), and the difference in weight between the largest and smallest nonrunt fish (90% CI, 0.0002 to 1.24). Additional observations indicated a possible protective effect attributable to previous exposure to the viruses. CONCLUSIONS AND CLINICAL RELEVANCE: Mortality and proportion of runts for white sturgeon after exposure to WSIV and WSHV-2 may be reduced for a farm at which the viruses are endemic by selection of specific broodstock, stocking with fish that survived outbreaks of viral disease, using all-in, all-out production, and decreasing stocking densities. PMID- 11039552 TI - Effects of xylazine hydrochloride during isoflurane-induced anesthesia in horses. AB - OBJECTIVE: To quantitate dose- and time-related anesthetic-sparing effects of xylazine hydrochloride (XYL) during isoflurane-induced anesthesia in horses and to characterize selected physiologic responses of anesthetized horses to administration of XYL. ANIMALS: 6 healthy adult horses. PROCEDURE: Horses were anesthetized 2 times to determine the minimum alveolar concentration (MAC) of isoflurane in O2 and to characterize the anesthetic-sparing effect (MAC reduction) after IV administration of XYL (0.5 and 1 mg/kg of body weight, random order). Selected measures of cardiopulmonary function, blood glucose concentrations, and urinary output also were measured during the anesthetic studies. RESULTS: Isoflurane MAC (mean +/- SEM) was reduced by 24.8 +/- 0.5 and 34.2 +/- 1.9% at 42 +/- 7 and 67 +/- 10 minutes, respectively, after administration of XYL at 0.5 and 1 mg/kg. Amount of MAC reduction by XYL was dose and time-dependent. Overall, cardiovascular and respiratory values varied little among treatments. Administration of XYL increased blood glucose concentration; the magnitude of change was dose- and time-dependent. Urine volume increased but not significantly. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of XYL reduced the anesthetic requirement for isoflurane in horses. The magnitude of the decrease is dose- and time-dependent. Administration of XYL increases blood glucose concentration in anesthetized horses in a dose-related manner. PMID- 11039554 TI - Effect of all-trans and 9-cis retinoic acid on growth and metastasis of xenotransplanted canine osteosarcoma cells in athymic mice. AB - OBJECTIVE: To determine effects of all-trans and 9-cis retinoic acid (RA) on tumor growth and metastatic ability of canine osteosarcoma cells transplanted into athymic (nude) mice. ANIMALS: Forty-five 5-week-old female BALB/c nude mice. PROCEDURE: 1 X 10(7) POS osteosarcoma cells were transplanted subcutaneously into the intrascapular region of mice. All-trans RA (3 or 30 microg/kg of body weight in 0.1 ml of sesame oil), 9-cis RA (3 or 30 mg/kg in 0.1 ml of sesame oil), or sesame oil (0.1 ml; control treatment) were administered intragastrically 5 d/wk for 4 weeks beginning 3 days after transplantation (n = 4 mice/group) or after formation of a palpable tumor (5 mice/group). Tumor weight was estimated weekly by measuring tumor length and width, and retinoid toxic effects were evaluated daily. Two weeks after the final treatment, mice were euthanatized, and number of mice with pulmonary metastases was determined. RESULTS: Adverse treatment effects were not detected. Tumor weight was less in mice treated with either dose of 9 cis RA than in control mice, although this difference was not significant. Treatment with 30 mg of 9-cis RA/kg initiated after tumor formation significantly reduced the incidence of pulmonary metastasis, compared with the control group. CONCLUSIONS AND CLINICAL RELEVANCE: 9-cis RA decreased the incidence of pulmonary metastasis in nude mice transplanted with canine osteosarcoma cells and may be a potential adjunct therapy for treatment of osteosarcoma in dogs. PMID- 11039555 TI - Regional anesthesia of the infraorbital and inferior alveolar nerves during noninvasive tooth pulp stimulation in halothane-anesthetized cats. AB - OBJECTIVE: To determine whether anesthesia of the infraorbital and inferior alveolar nerves abolishes reflex-evoked muscle action potentials (REMP) during tooth-pulp stimulation in halothane-anesthetized cats. ANIMALS: 8 healthy adult cats. PROCEDURE: In halothane-anesthetized cats, an anodal electrode was attached to the tooth to be stimulated and a platinum needle cathodal electrode was inserted in adjacent gingival mucosa. Cathodal and anodal electrodes were moved to the upper and lower canine, upper fourth premolar, and lower first molar teeth for stimulation; baseline REMP was recorded. A 25-gauge 1-cm needle was inserted 0.5 cm into the infraorbital canal. A 25-gauge 1-cm needle was inserted 1 cm rostral to the angular process of the ramus, and advanced 0.5 cm along the medial aspect. Chloroprocaine was injected at each site. Each tooth was stimulated every 10 minutes for 90 minutes. RESULTS: REMP was abolished within 10 minutes for all upper teeth, except for the upper canine tooth in 1 cat, and abolished within 10 minutes for lower teeth in 4 cats. In 1 cat, REMP was not abolished in the lower first molar tooth. In 3 cats, REMP was not abolished in the lower canine and first molar teeth. At 90 minutes, REMP was restored for all teeth except the lower canine tooth in 1 cat, for which REMP was restored at 120 minutes. CONCLUSIONS AND CLINICAL RELEVANCE: Regional anesthesia of the infraorbital and inferior alveolar nerves may provide dental analgesia in cats. PMID- 11039556 TI - Platelet aggregation in dogs with mitral valve regurgitation. AB - OBJECTIVE: To compare platelet aggregation in healthy dogs and dogs with mitral valve regurgitation (MVR) to determine whether regurgitation had an effect on platelet function. ANIMALS: 32 dogs with MVR and 43 healthy dogs. PROCEDURE: Platelet aggregation was measured with an aggregometer, using adenosine 5' diphosphate as the aggregating agent, and the maximum aggregation and the enhancement of platelet sensitivity (EPS) values were calculated. RESULTS: Platelet count and maximum aggregation were not significantly different between healthy dogs and dogs with MVR. However, EPS values in dogs with MVR were significantly higher than values in healthy dogs. Platelet count and maximum aggregation were not significantly different between dogs classified as New York Heart Association functional class I or II and dogs classified as functional class III or IV; however, EPS values were significantly higher in dogs classified as functional class III or IV. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that platelet aggregation is decreased in dogs with MVR and that the EPS value may be more sensitive to differences in disease severity than in measurement of maximum aggregation. PMID- 11039557 TI - Clinical effects of exercise on subchondral bone of carpal and metacarpophalangeal joints in horses. AB - OBJECTIVE: To determine effects of treadmill exercise on subchondral bone of carpal and metacarpophalangeal joints of 2-year-old horses. ANIMALS: 12 healthy 2 year-old horses. PROCEDURE: Horses were randomly assigned to the control (n = 6) or exercised (6) groups. Horses in the exercised group ran on a high-speed treadmill 5 d/wk for 6 months. Horses in the control group were hand walked for the same amount of time. Results of clinical, radiographic, nuclear scintigraphic, and computed tomographic examinations, and serum and synovial concentrations of biochemical markers of bone metabolism were compared between groups. RESULTS: Exercised horses were significantly lamer at the end of the study than control horses. Radionuclide uptake in the metacarpal condyles, but not in the carpal joints, was greater in exercised horses, compared with control horses. Exercised horses also had a higher subchondral bone density in the metacarpal condyles than control horses, but such differences were not detected in the carpal bones. CONCLUSIONS AND CLINICAL RELEVANCE: None of the diagnostic techniques evaluated was sufficiently sensitive to detect all osteochondral damage. Computed tomography and computed tomographic osteoabsorptiometry were superior to conventional radiography for detecting small osteochondral fragments. Nuclear scintigraphy was a sensitive indicator of subchondral bone change but lacked specificity for describing lesions and discerning normal bone remodeling from damage. Newer techniques such as computed tomography may help clinicians better diagnose early and subtle joint lesions in horses prior to development of gross joint damage. PMID- 11039558 TI - In vitro investigation of the effect of prostaglandins and nonsteroidal anti inflammatory drugs on contractile activity of the equine smooth muscle of the dorsal colon, ventral colon, and pelvic flexure. AB - OBJECTIVES: To determine the in vitro effect of prostaglandin E2 (PGE2), PGF2alpha, PGI2; and nonsteroidal anti-inflammatory drugs (NSAID; ie, flunixin meglumine, ketoprofen, carprofen, and phenylbutazone) on contractile activity of the equine dorsal colon, ventral colon, and pelvic flexure circular and longitudinal smooth muscle. ANIMALS: 26 healthy horses. PROCEDURE: Tissue collected from the ventral colon, dorsal colon, and pelvic flexure was cut into strips and mounted in a tissue bath system where contractile strength was determined. Incremental doses of PGE2, PGF2alpha,, PGI2, flunixin meglumine, carprofen, ketoprofen, and phenylbutazone were added to the baths, and the contractile activity was recorded for each location and orientation of smooth muscle. RESULTS: In substance P-stimulated tissues, PGE2 and PGF2alpha enhanced contractility in the longitudinal smooth muscle with a decrease or no effect on circular smooth muscle activity. Prostaglandin I2 inhibited the circular smooth muscle response with no effect on the longitudinal muscle. The activity of NSAID was predominantly inhibitory regardless of location or muscle orientation. CONCLUSIONS AND CLINICAL RELEVANCE: In the equine large intestine, exogenous prostaglandins had a variable effect on contractile activity, depending on the location in the colon and orientation of the smooth muscle. The administration of NSAID inhibited contractility, with flunixin meglumine generally inducing the most profound inhibition relative to the other NSAID evaluated in substance P stimulated smooth muscle of the large intestine. The results of this study indicate that prolonged use of NSAID may potentially predispose horses to develop gastrointestinal tract stasis and subsequent impaction. PMID- 11039559 TI - Histomorphometric analysis of the proximal portion of the femur in dogs with osteoarthritis. AB - OBJECTIVE: To describe cancellous architecture of the proximal portion of the femur in dogs with osteoarthritis. ANIMALS: 30 dogs with coxofemoral osteoarthritis. PROCEDURE: All dogs had femoral head and neck excision or total hip arthroplasty. Histomorphometry software was used to analyze computer images of 100-microm-thick coronal and transverse plane sections of the head and neck of the femur. Histologic preparations of coronal and transverse sections of articular cartilage were graded. RESULTS: Bone volume/total volume, trabecular thickness, trabecular number, and bone surface/total volume were significantly higher in the femoral head than femoral neck. Trabecular alignment (anisotropy) and separation were significantly higher in the femoral neck than femoral head. Anisotropy was significantly increased in the medial portion of the femoral head in the coronal plane and in the cranial portion of the femoral neck in the transverse plane, compared with healthy dogs. The medial half of femoral head cartilage that overlies the proximomedial cancellous bone region had significantly more degraded cartilage than the lateral half. Histologic grades for cranial and caudal halves of femoral head articular cartilage were similar. CONCLUSIONS AND CLINICAL RELEVANCE: Most findings were similar to those in healthy dogs. Greater trabecular alignment in the proximomedial region of the femoral head and craniolateral region of the femoral neck in dogs with osteoarthritis suggests an altered transfer of load through the coxofemoral joint. Greater cartilage degradation on the medial half of the femoral head supports an association between increased trabecular alignment and cartilage degradation. PMID- 11039560 TI - Effects of anesthesia, surgery, and intravenous administration of fluids on plasma antidiuretic hormone concentrations in healthy dogs. AB - OBJECTIVE: To evaluate effects of anesthesia, surgery, and intravenous administration of fluids on plasma concentrations of antidiuretic hormone (ADH), concentration of total solids (TS), PCV, arterial blood pressure (BP), plasma osmolality, and urine output in healthy dogs. ANIMALS: 22 healthy Beagles. PROCEDURE: 11 dogs did not receive fluids, and 11 received 20 ml of lactated Ringer's solution/kg of body weight/h. Plasma ADH adn TS concentrations, PCV, osmolality, and arterial BP were measured before anesthesia (T0) and after administration of preanesthetic agents (T1), induction of anesthesia (T2), and 1 and 2 hours of surgery (T3 and T4, respectively). Urine output was measured at T3 and T4. RESULTS: ADH concentrations increased at T1, T3, and T4, compared with concentrations at T0. Concentration of TS and PCV decreased at all times after administration of preanesthetic drugs. Plasma ADH concentration was less at T3 in dogs that received fluids, compared with those that did not. Blood pressure did not differ between groups, and osmolality did not increase > 1% from To value at any time. At T4, rate of urine production was less in dogs that did not receive fluids, compared with those that did. CONCLUSIONS AND CLINICAL RELEVANCE: Plasma ADH concentration increased and PCV and TS concentration decreased in response to anesthesia and surgery. Intravenous administration of fluids resulted in increased urine output but had no effect on ADH concentration or arterial BP. The causes and effects of increased plasma ADH concentrations may affect efficacious administration of fluids during the perioperative period in dogs. PMID- 11039561 TI - Mutational analysis of tumor suppressor gene p53 in feline vaccine site associated sarcomas. AB - OBJECTIVES: To investigate the role of tumor suppressor gene p53 mutation in feline vaccine site-associated sarcoma (VSS) development and to evaluate the relationship between p53 nucleotide sequence and protein expression. SAMPLE POPULATION: Formalin-fixed paraffin-embedded tissues of 8 feline VSS with dark p53 immunostaining (high p53 expression) and 13 feline VSS with faint or no staining (normal p53 expression). PROCEDURE: DNA was extracted from neoplastic and normal tissue from each paraffin block. The following 3 regions of the p53 gene were amplified by polymerase chain reaction: 379 base pair (bp) region of exon 5, intron 5, and exon 6, 108 bp region of exon 7, and 140 bp region of exon 8. Amplified p53 products were sequenced and compared with published feline p53. The p53 mutations identified were correlated with p53 mutations predicted by immunostaining. RESULTS: Neoplastic cells of 5 of 8 (62.5%) VSS that had high p53 expression harbored single missense mutations within the p53 gene regions examined. The p53 gene mutations were not detected in the 13 tumors with normal p53 immunostaining. Nonneoplastic tissues adjacent to all 21 VSS lacked mutations of these p53 gene regions. CONCLUSIONS: The p53 gene mutations were restricted to neoplastic tissue and, therefore, were unlikely to predispose to VSS. However, p53 mutations may have contributed to cancer progression in 5 of the 21 VSS. There was very good (kappa quotient = 0.67 with a confidence limit of 0.3 to 1.0), although not complete, agreement between prediction of mutation by p53 immunostaining and identification of mutations by sequencing of key p53 gene regions. PMID- 11039562 TI - Effect of a 30-minute infusion of dobutamine hydrochloride on hind limb blood flow and hemodynamics in halothane-anesthetized horses. AB - OBJECTIVE: To evaluate the hemodynamic effects of dobutamine hydrochloride (0.5 microg/kg of body weight/min) in halothane-anesthetized horses. ANIMALS: 6 adult Thoroughbred horses. PROCEDURE: Anesthesia was induced by use of romifidine (100 microg/kg) and ketamine (2.2 mg/kg), IV. Anesthesia was maintained by halothane (end-tidal concentration 0.9 to 1.0%). Aortic, left ventricular, and right atrial pressures were measured, using catheter-mounted strain gauge transducers. Cardiac output (CO), velocity time integral, maximal aortic blood flow velocity and acceleration, and left ventricular preejection period and ejection time were measured from aortic velocity waveforms obtained by transesophageal Doppler echocardiography. Velocity waveforms were recorded from the femoral vessels, using Doppler ultrasonography. The time-averaged mean velocity and early diastolic deceleration slope (EDDS) were measured. Pulsatility index (PI) and volumetric flow were calculated. Microvascular perfusion was measured in the semimembranosus muscles by laser Doppler flowmetry. Data were recorded 60 minutes after induction of anesthesia (control) and at 15 and 30 minutes after start of an infusion of dobutamine (0.5 microg/kg/min). RESULTS: Aortic pressures were significantly increased during the infusion of dobutamine. No change was observed in the indices of left ventricular systolic function including CO. Femoral arterial flow significantly increased, and the PI and EDDS decreased. No change was observed in the femoral venous flow or in microvascular perfusion. CONCLUSIONS AND CLINICAL RELEVANCE: At this dosage, dobutamine did not alter left ventricular systolic function. Femoral blood flow was preferentially increased as the result of local vasodilatation. The lack of effect of dobutamine on microvascular perfusion suggests that increased femoral flow is not necessarily associated with improved perfusion of skeletal muscles. PMID- 11039563 TI - Comparison of tear proteins of llamas and cattle. AB - OBJECTIVE: To analyze and compare contents of the preocular tear films of llamas and cattle. ANIMALS: 40 llamas and 35 cattle. PROCEDURE: Tear pH was determined by use of a pH meter. Total protein concentration was determined by use of 2 microtiter methods. Tear proteins were separated by use of electrophoresis and molecular weights of bands were calculated. Western blot immunoassay was used to detect IgA, lactoferrin, transferrin, ceruloplasmin, alpha1-antitrypsin, alpha1 amylase, and alpha2-macroglobulin. Enzyme electrophoresis was used to detect proteases. RESULTS: The pH of llama and cattle tears were 8.05 +/- 0.01 and 8.10 +/- 0.01, respectively. For results of both methods, total protein concentration of llama tears was significantly greater than that of cattle tears. Molecular weights of tear protein bands were similar within and between the 2 species, although llama tears had a distinct 13.6-kd band that was not detected in cattle. Lactoferrin, IgA, transferrin, ceruloplasmin, alpha1-antitrypsin, alpha1-amylase, alpha2-macroglobulin, and proteases were detected in both species. CONCLUSIONS AND CLINICAL RELEVANCE: Llama tears have significantly greater total protein concentration than cattle tears, whereas pH is similar between species. Because little variation was detected within species for the number and molecular weight of protein bands, pooling of tears for analysis is justified. Results suggest that lactoferrin, ceruloplasmin, transferrin, alpha1-antitrypsin, alpha2 macroglobulin, alpha1-amylase, and IgA are present in the tears of llamas and cattle. PMID- 11039564 TI - Detection of lysozyme in llama, sheep, and cattle tears. AB - OBJECTIVE: To determine whether the tears of llamas, sheep, and cattle contain lysozyme and compare lysozyme concentrations in tears among these species. ANIMALS: 40 llamas, 5 sheep, and 36 cattle. PROCEDURE: Electrophoresis, western blot immunoassay for lysozyme, a spectrophotometric assay to detect tear lysozyme by its ability to lyse a suspension of Micrococcus lysodeiticus, and a microtiter plate colorometric assay were performed. RESULTS: A 13.6-kd protein band was detected by use of electrophoresis and western blot immunoassay in llama and sheep tears but not cattle tears. Results of spectrophotometric assay suggested that llama and sheep tears had high concentrations of lysozyme, whereas cattle tears had low concentrations. Results of the microtiter plate colorometric assay suggested that llama tears had high concentrations of lysozyme, whereas concentrations in sheep and cattle tears were lower. CONCLUSIONS AND CLINICAL RELEVANCE: Lysozyme concentrations in tears may vary among species and this variability may contribute to differing susceptibilities to ocular diseases such as infectious keratoconjunctivitis. PMID- 11039565 TI - In vitro comparison of the use of two large-animal, centrally threaded, positive profile transfixation pin designs in the equine third metacarpal bone. AB - OBJECTIVE: To compare the in vitro holding power and associated microstructural damage of 2 large-animal centrally threaded positive-profile transfixation pins in the diaphysis of the equine third metacarpal bone. SAMPLE POPULATION: 25 pairs of adult equine cadaver metacarpal bones. PROCEDURE: Centrally threaded positive profile transfixation pins of 2 different designs (ie, self-drilling, self tapping [SDST] vs nonself-drilling, nonself-tapping [NDNT] transfixation pins) were inserted into the middiaphysis of adult equine metacarpal bones. Temperature of the hardware was measured during each step of insertion with a surface thermocouple. Bone and cortical width, transfixation pin placement, and cortical damage were assessed radiographically. Resistance to axial extraction before and after cyclic loading was measured using a material testing system. Microstructural damage caused by transfixation pin insertion was evaluated by scanning electron microscopy. RESULTS: The temperature following pin insertion was significantly higher for SDST transfixation pins. Periosteal surface cortical fractures were found in 50% of the bones with SDST transfixation pins and in none with NDNT transfixation pins. The NDNT transfixation pins were significantly more resistant to axial extraction than SDST transfixation pins. Grossly and microscopically, NDNT transfixation pins created less damage to the bone and a more consistent thread pattern. CONCLUSIONS AND CLINICAL RELEVANCE: In vitro analysis revealed that insertion of NDNT transfixation pins cause less macroscopic and microscopic damage to the bone than SDST transfixation pins. The NDNT transfixation pins have a greater pull out strength, reflecting better initial bone transfixation pin stability. PMID- 11039566 TI - In vitro comparison of metaphyseal and diaphyseal placement of centrally threaded, positive-profile transfixation pins in the equine third metacarpal bone. AB - OBJECTIVE: To evaluate in vitro holding power and associated microstructural and thermal damage from placement of positive-profile transfixation pins in the diaphysis and metaphysis of the equine third metacarpal bone. SAMPLE POPULATION: Third metacarpal bones from 30 pairs of adult equine cadavers. PROCEDURE: Centrally threaded positive-profile transfixation pins were placed in the diaphysis of 1 metacarpal bone and the metaphysis of the opposite metacarpal bone of 15 pairs of bones. Tensile force at failure for axial extraction was measured with a materials testing system. An additional 15 pairs of metacarpal bones were tested similarly following cyclic loading. Microstructural damage was evaluated via scanning electron microscopy in another 6 pairs of metacarpal bones, 2 pairs in each of the following 3 groups: metacarpal bones with tapped holes and without transfixation pin placement, metacarpal bones following transfixation pin placement, and metacarpal bones following transfixation pin placement and cyclic loading. Temperature of the hardware was measured with a surface thermocouple in 12 additional metacarpal bones warmed to 38 C. RESULTS: The diaphysis provided significantly greater resistance to axial extraction than the metaphysis. There were no significant temperature differences between diaphyseal and metaphyseal placement. Microstructural damage was limited to occasional microfractures seen only in cortical bone of diaphyseal and metaphyseal locations. Microfractures originated during drilling and tapping but did not worsen following transfixation pin placement or cyclic loading. CONCLUSIONS AND CLINICAL RELEVANCE: Centrally threaded, positive-profile transfixation pins have greater resistance to axial extraction in the diaphysis than in the metaphysis of equine third metacarpal bone in vitro. This information may be used to create more stable external skeletal fixation in horses with fractures. PMID- 11039568 TI - In vitro determination of contact areas in the normal elbow joint of dogs. AB - OBJECTIVE: To evaluate areas of articular contact of the proximal portions of the radius and ulna in normal elbow joints of dogs and the effects of axial load on size and location of these areas. SAMPLE POPULATION: Forelimbs obtained from cadavers of 5 adult mixed-breed dogs. PROCEDURE: After forelimbs were removed, liquid-phase polymethyl methacrylate was applied to articular surfaces of the elbow joint, and limbs were axially loaded. Articular regions void of casting material were stained with water-soluble paint. Relative articular contact areas were determined by computer-assisted image analyses of stained specimens. Repeatability of the technique was evaluated by analyses of casts from bilateral forelimbs of 1 cadaver. Incremental axial loads were applied to left forelimbs from 4 cadavers to determine effects of load on articular contact. RESULTS: Specific areas of articular contact were identified on the radius, the craniolateral aspect of the anconeus, and the medial coronoid process. The medial coronoid and radial contact areas were continuous across the radioulnar articulation. There was no articular contact of the medial aspect of the anconeus with the central trochlear notch. Coefficients of variation of contact areas between repeated tests and between contralateral limbs was < 20%. Significant overall effects of axial load on contact area or location were not identified. CONCLUSIONS: Three distinct contact areas were evident in the elbow joint of dogs. Two ulnar contact areas were detected, suggesting there may be physiologic incongruity of the humeroulnar joint. There was no evidence of surface incongruity between the medial edge of the radial head and the lateral edge of the medial coronoid process. PMID- 11039567 TI - Effects of 3-methylindole production and immunity against bovine respiratory syncytial virus on development of respiratory tract disease and rate of gain of feedlot cattle. AB - OBJECTIVE: To determine whether immunity against bovine respiratory syncytial virus (BRSV) mitigates the effects of 3-methylindole (3MI) on occurrence of bovine respiratory tract disease (BRD) and rate of gain in feedlot cattle. ANIMALS: 254 mixed-breed beef cattle. PROCEDURE: Cattle were randomly assigned to 1 of 3 groups at the time of arrival at the feedlot. One group was vaccinated with an inactivated BRSV vaccine, another was vaccinated with a modified-live BRSV vaccine, and the third was maintained as unvaccinated control cattle. On days 0 and 28, serum BRSV antibody concentrations were measured, using serum neutralizing and ELISA techniques. Serum 3MI concentrations were measured at feedlot arrival and 3 days later. Cattle were monitored for development of BRD. At slaughter, lungs were evaluated grossly for chronic lesions. RESULTS: Higher serum 3MI concentrations early in the feeding period were associated with lower mean daily gain. Control cattle were more likely to be treated for BRD after day 3, compared with cattle vaccinated with the modified-live BRSV vaccine. Humoral immunity against BRSV did not appear to modify the effect of 3MI on development of BRD or mean daily gain. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that abrogating the effects of 3MI and BRSV infection may improve the health and growth performance of feedlot cattle. However, in this study, immunity against BRSV did not appear to protect against the potential synergism between 3MI and BRSV infection, possibly because of the slow rates of gain of cattle included in the study or timing of sample collection. PMID- 11039569 TI - Activities of NADPH-dependent reductases and sorbitol dehydrogenase in canine and feline lenses. AB - OBJECTIVE: To measure activities of NADPH-dependent reductases and sorbitol dehydrogenase in lenses from healthy dogs and cats. SAMPLE POPULATION: Lenses from 37 dogs and 23 cats. All animals were healthy and had serum glucose concentrations within reference limits. PROCEDURE: Lenses were homogenized, and activities of NADPH-dependent reductases and sorbitol dehydrogenase were measured spectrophotometrically. RESULTS: Activities of NADPH-dependent reductases and sorbitol dehydrogenase were significantly lower in lenses from cats than in lenses from dogs. However, the ratio of NADPH-dependent reductases activity-to sorbitol dehydrogenase activity was significantly higher in lenses from cats than in lenses from dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that during periods of hyperglycemia, sorbitol would accumulate at a faster rate in the lenses of cats than in the lenses of dogs. Thus, the higher incidence of diabetic cataracts in dogs, compared with cats, is likely not attributable to a difference in the ratio of NADPH-dependent reductases activity-to-sorbitol dehydrogenase activity. PMID- 11039570 TI - Determinants of oxygen delivery and hemoglobin saturation during incremental exercise in horses. AB - OBJECTIVE: To determine components of the increase in oxygen consumption (VO2) and evaluate determinants of hemoglobin saturation (SO2) during incremental treadmill exercise in unfit horses. ANIMALS: 7 unfit adult mares. PROCEDURES: Horses performed 1 preliminary exercise test (EXT) and 2 experimental EXT. Arterial and mixed venous blood samples and hemodynamic measurements were taken during the last 30 seconds of each step of the GXT to measure PO2, hemoglobin concentration ([Hb]), SO2, and determinants of acid-base state (protein, electrolytes, and PCO2). RESULTS: Increased VO2 during exercise was facilitated by significant increases in cardiac output (CO), [Hb], and widening of the arteriovenous difference in O2. Arterial and venous pH, PaO2, and PvO2 decreased during exercise. Arterial PCO2, bicarbonate ([HCO3-])a, and [HCO3-] decreased significantly, whereas PVCO2 and increased. Arterial and venous sodium concentration, potassium concentration, strong ion difference, and venous lactate concentration all increased significantly during exercise. CONCLUSIONS AND CLINICAL RELEVANCE: Increases in CO, [Hb], and O2 extraction contributed equally to increased VO2 during exercise. Higher PCO2 did not provide an independent contribution to shift in the oxyhemoglobin dissociation curve (OCD) in venous blood. However, lower PaCO2 shifted the curve leftward, facilitating O2 loading. The shift of ODC resulted in minimal effect on O2 extraction because of convergence of the ODC at lower values of PO2. Decreased pH appeared responsible for the rightward shift of the ODC, which may be necessary to allow maximal O2 extraction at high blood flows achieved during exercise. PMID- 11039571 TI - Psychological consequences of predictive genetic testing: a systematic review. AB - The aim of this systematic literature review is to describe the psychological consequences of predictive genetic testing. Five databases were searched for studies using standardised outcome measures and statistical comparison of groups. Studies were selected and coded by two independent researchers. From 899 abstracts, 15 papers, describing 11 data sets, met the selection criteria for the review. The studies were of predictive genetic testing for Huntington's disease, hereditary breast and ovarian cancer, familial adenomatous polyposis and spinocerebellar ataxia. One involved children; the rest were of adults. None of the 15 papers reported increased distress (general and situational distress, anxiety and depression) in carriers or non-carriers at any point during the 12 months after testing. Both carriers and non-carriers showed decreased distress after testing; this was greater and more rapid amongst non-carriers. Test result (ie being a carrier or non-carrier) was rarely predictive of distress more than one month after testing (predictive in two of 14 analyses). Pre-test emotional state was predictive of subsequent distress in 14 of 27 analyses. There is a lack of informative studies in this field. The studies reviewed suggest that those undergoing predictive genetic testing do not experience adverse psychological consequences. However, the studies are of self-selected populations who have agreed to participate in psychological studies and have been followed up for no more than three years. Most research has been of testing for Huntington's Disease and included follow-up of no more than one year. The results suggest that testing protocols should include a pre-test assessment of emotional state so that post test counselling can be targeted at those more distressed before testing. None of the studies experimentally manipulated the amount or type of counselling provided. The relationship between counselling and emotional outcome is therefore unclear and awaits empirical study. PMID- 11039572 TI - Protection of privacy by third-party encryption in genetic research in Iceland. AB - As the new human genetics continues its dramatic expansion into many laboratories and medical institutions, the concern for the protection of the personal privacy of individuals who participate increases. It seems that even the smallest of laboratories must confront the issue of how to protect the genetic and phenotypic information of participants in their research. Some have promoted the use of anonymity as a way out of this dilemma. But we are reminded by others that the future cannot be predicted, and that future benefits may be lost when the links to these benevolent volunteers are gone forever. More recently, some ethical bodies have suggested, without specific recommendations, that a reversible third party encryption system may be a solution to this problem. However, they have not provided a route or even examples of how to proceed. We present here the Icelandic approach to this issue by developing a third-party encryption system in direct collaboration with the Data Protection Commission (DPC) of Iceland. We have incorporated the encryption system within our sample collection and storage software, which minimises inconvenience but enhances security. The strategy assures a barrier between the laboratory and the outside world that can only be crossed by the DPC. PMID- 11039573 TI - Inheritance of human longevity in Iceland. AB - The idea that human longevity is influenced by genetic factors has recently received strong support from work on other species. On the basis of partial population studies and selected kinships, significant correlations between the ages of parents and offspring have been reported, and some but not all twin studies have confirmed that human longevity is moderately inherited. However, studies based upon a relatively small proportion of a population are susceptible to sampling error and selection bias. Here we report the use of a comprehensive population-based computerised genealogy database to examine multigenerational relationships among those who live to the 95th percentile in Iceland. We have developed a clustering tool which can generate large extended pedigrees connecting individuals from any list using the genealogy database. First degree relatives of those living to the 95th percentile are almost twice as likely to live to the 95th percentile compared with controls. Furthermore, we have developed an algorithm which we have named the Minimum Founder Test (MFT) to examine the degree of relatedness of any population-based list of individuals to estimate whether a trait has a familial component. The data indicate that there is a significant genetic component to longevity. In addition, age-specific death rates are significantly lower in the offspring of long-lived parents compared with controls, especially after age 70. PMID- 11039574 TI - Inferring the impact of linguistic boundaries on population differentiation: application to the Afro-Asiatic-Indo-European case. AB - We present here a quantitative way to assess the impact of language-family boundaries on population differentiation and to evaluate the homogeneity of the genetic processes along these boundaries. Our estimator (delta a) of the impact of the boundary is based on an isolation by distance (IBD) model and measures the added genetic distance between populations located on different sides of the boundary. We compare this statistic with another estimator of group differentiation (F(CT)) computed under an analysis of variance framework that does not assume any particular spatial structure of the populations. Monte Carlo simulations are used to study the behaviour of these statistics under a two dimensional stepping-stone model. Simulations show that F(CT) can suggest the existence of a frontier when populations only differ because of IBD. This spurious behaviour is much less frequent for the delta a statistic. However, the large variance associated with the delta a statistic, and the fact that it should only be computed in the presence of IBD, may limit the use of this statistic. Overall, the origin and the effect of the boundary is best understood by comparing different statistics and by testing for the presence of IBD on each side of the boundary as well as across the boundary. We illustrate our approach by examining the boundary between Afro-Asiatic and Indo-European populations. These populations are globally genetically differentiated, but the effect of the linguistic boundary on gene flow seems geographically very heterogeneous. This boundary appears to be the result of a secondary contact between two differentiation centres rather than an enhancer of population differentiation. PMID- 11039575 TI - Multiple founder effects and geographical clustering of BRCA1 and BRCA2 families in Finland. AB - In the Finnish breast and ovarian cancer families six BRCA1 and five BRCA2 mutations have been found recurrently. Some of these recurrent mutations have also been seen elsewhere in the world, while others are exclusively of Finnish origin. A haplotype analysis of 26 Finnish families carrying a BRCA1 mutation and 20 families with a BRCA2 mutation indicated that the carriers of each recurrent mutation have common ancestors. The common ancestors were estimated to trace back to 7-36 generations (150-800 years). The time estimates and the geographical clustering of these founder mutations in Finland are in concordance with the population history of this country. Analysis of the cancer phenotypes showed differential ovarian cancer expression in families carrying mutations in the 5' and 3' ends of the BRCA1 gene, and earlier age of ovarian cancer onset in families with BRCA1 mutations compared with families with BRCA2 mutations. The identification of prominent and regional BRCA1 and BRCA2 founder mutations in Finland will have significant impact on diagnostics in Finnish breast and ovarian cancer families. An isolated population with known history and multiple local founder effects in multigenic disease may offer distinct advantages also for mapping novel predisposing genes. PMID- 11039576 TI - FISHing for mechanisms of cytogenetically defined terminal deletions using chromosome-specific subtelomeric probes. AB - Cytogenetically defined terminal deletions are thought to be a major, yet underappreciated, cause of mental retardation and multiple congenital anomalies. The mechanisms by which terminal deletions arise and are stabilized are not completely understood; although all ends of human chromosomes must have a telomeric cap to be stable. At least three mechanisms exist to maintain chromosome ends with cytogenetically defined terminal deletions: stabilization of terminal deletions through a process of telomere regeneration (termed 'telomere healing'), retention of the original telomere producing interstitial deletions, and formation of derivative chromosomes by obtaining a different telomeric sequence through cytogenetic rearrangement (termed 'telomere capture'). We used chromosome-specific subtelomeric probes and FISH to characterize cytogenetically defined terminal deletions in patients with 1p36 monosomy. Based on the current resolution of these subtelomeric probes, our results indicate that cytogenetically defined terminal deletions of 1p36 are likely to occur through all three mechanisms, although we speculate that the majority of cases were stabilized through telomere regeneration. These results demonstrate the use of chromosome-specific subtelomeric probes as an efficient first step toward uncovering the mechanisms that result in the stabilization of cytogenetically defined terminal deletions. PMID- 11039577 TI - Hereditary spastic paraplegia caused by mutations in the SPG4 gene. AB - Autosomal dominant hereditary spastic paraplegia (AD-HSP) is a genetically heterogeneous neurodegenerative disorder characterised by progressive spasticity of the lower limbs. The SPG4 locus at 2p21-p22 accounts for 40-50% of all AD-HSP families. The SPG4 gene was recently identified. It is ubiquitously expressed in adult and foetal tissues and encodes spastin, an ATPase of the AAA family. We have now identified four novel SPG4 mutations in German AD-HSP families, including one large family for which anticipation had been proposed. Mutations include one frame-shift and one missense mutation, both affecting the Walker motif B. Two further mutations affect two donor splice sites in introns 12 and 16, respectively. RT-PCR analysis of both donor splice site mutations revealed exon skipping and reduced stability of aberrantly spliced SPG4 mRNA. All mutations are predicted to cause loss of functional protein. In conclusion, we confirm in German families that SPG4 mutations cause AD-HSP. Our data suggest that SPG4 mutations exert their dominant effect not by gain of function but by haploinsufficiency. If a threshold level of spastin were critical for axonal preservation, such threshold dosage effects might explain the variable expressivity and incomplete penetrance of SPG4-linked AD-HSP. PMID- 11039578 TI - A refined physical and transcriptional map of the SPG9 locus on 10q23.3-q24.2. AB - Hereditary spastic paraplegia (HSP) is a genetically heterogeneous disorder characterised by progressive spasticity of the lower limbs. Beside 'pure' forms of HSP, 'complicated' forms are reported, where spasticity occurs associated with additional symptoms. We recently described an Italian family with a complicated dominant form of HSP (SPG9) and we mapped the gene responsible to 10q23.3-q24.2, in a 12cM interval between markers D10S564 and D10S603. The phenotypic manifestations in our family are reminiscent of those already described in a smaller British pedigree. We typed individuals from this British family using markers located in the SPG9 critical interval and haplotype reconstruction showed the disorder co-segregating with SPG9. To characterise the SPG9 region better, we constructed a contig of 22 YACs, assigned it to 18 polymorphic markers and positioned 54 ESTs. Furthermore, we searched for ESTs containing a trinucleotide repeat sequence, since anticipation of symptoms was reported in both families. Finally, analysis of a muscle biopsy specimen from one patient was normal, suggesting that, contrary to SPG7, mitochondrial disturbance could not be a primary feature of SPG9. PMID- 11039579 TI - NRL S50T mutation and the importance of 'founder effects' in inherited retinal dystrophies. AB - The aim of this work was to identify NRL mutations in a panel of 200 autosomal dominant retinitis pigmentosa (adRP) families. All samples were subjected to heteroduplex analysis of the three exons of the NRL gene, and HphI restriction digest analysis of exon 2 (to identify the S50T mutation). Families found to have the S50T mutation, and six additional larger pedigrees (which had previously been excluded from the other nine adRP loci) underwent linkage analysis using polymorphic markers located in the region of 14q11. HphI restriction analysis followed by direct sequencing of the amplified NRL exon 2 product demonstrated the presence of the NRL S50T sequence change in three adRP families. Comparison of marker haplotypes in affected individuals from these families with those of affected members of the original 14q11 linked family revealed a common disease haplotype for markers within the adRP locus. Recombination events observed in these families define an adRP critical interval of 14.9 cM between D13S72 and D14S1041. Linkage analysis enabled all six of the larger adRP pedigrees to be excluded from the 14q11 locus. The NRL S50T mutation represents another example of a 'founder effect' in a dominantly inherited retinal dystrophy. Identification of such 'founder effects' may greatly simplify diagnostic genetic screening and lead to better prognostic counselling. The exclusion of several adRP families from all ten adRP loci indicates that at least one further adRP locus remains to be found. PMID- 11039580 TI - The IL9R region contribution in asthma is supported by genetic association in an isolated population. AB - Interleukin 9 (IL9) is involved in mast cell maturation and the enhancement of IgE production by B cells. Furthermore, linkage data in human and mice have suggested that IL9 may contribute to asthma. Since our genetic analysis of the 5q cytokine cluster did not support a genetic role for the IL9 gene, we became interested in the IL9 receptor gene (IL9R) in the pseudoautosomal region. We genotyped markers sDF2 and sDF1 close to the IL9R gene among 289 affected and 368 family-based controls. The results were studied by using linkage, transmission disequilibrium, association and homozygosity analyses. Linkage analyses remained negative, presumably because of our low power for linkage study. However, all the other analyses yielded evidence that the IL9R gene region may have a role in the development of asthma. The sDF2*10 allele was more frequently transmitted than untransmitted to asthmatic offspring (34 vs 16, pchi2 < or = 0.01), and it was found homozygotic among asthma patients more often than expected (Psimul2 = 0.009). Also, a specific X chromosomal haplotype, sDF2*10-sDF1*6 associated with asthma (40 vs 7, Pchi2 < 0.005, Psimul1 = 0.04). PMID- 11039581 TI - Dystrophin nonsense mutation induces different levels of exon 29 skipping and leads to variable phenotypes within one BMD family. AB - Within one X-linked muscular dystrophy family, different phenotypes for three males occurred: (1) a severely affected Becker patient with cardiomyopathy, (2) a mildly affected Becker patient, and (3) an apparently healthy male with elevated serum CK levels. In the muscle biopsy specimen of patient2 one out of four antibodies (NCL-DYS1) showed absence of dystrophin. The protein truncation test detected a truncated dystrophin for both muscle tissue and lymphocytes of this patient next to an additional near normal size fragment in muscle. Genomic sequence analysis revealed a nonsense mutation in exon 29 (4148C > T) of the dystrophin gene. Sequence analysis of the mRNA fragment of the larger peptide showed skipping of exon 29, restoring an open reading frame. Consequently, the epitope of the antibody NCL-DYS1 is mapped to exon 29. The variable clinical features of the three relatives from healthy to severely affected therefore seems to be related to the level of skipping of exon 29. This finding underscores the future potential of gene therapeutic strategies aimed at inducing exon skipping in Duchenne muscular dystrophy, to generate a much milder disease. PMID- 11039582 TI - A new mutation in the six-domain of SIX3 gene causes holoprosencephaly. AB - Holoprosencephaly (HPE) is a severe brain malformation which results from incomplete cleavage of the forebrain during early embryogenesis. The aetiology of HPE is very heterogeneous. Among the genetic factors, SIX3, which is considered to be the functional orthologue of Drosophila genes sine oculis (so) and optix, has been found to be mutated in the homeodomain, in some patients with HPE (HPE2 on chromosome 2p21). We report a new HPE family, presenting a wide spectrum of clinical features, ranging from cyclopia to hypotelorism, in which a mutation was found for the first time in the SIX domain of SIX3: a GG insertion creates a frameshift leading to a nonsense mutation downstream in the homeodomain region. PMID- 11039583 TI - Inv dup(22), del(22)(q11) and r(22) in the father of a child with DiGeorge syndrome. AB - We here report a unique inherited case of DiGeorge syndrome. The asymptomatic father had a mosaic karyotype with a 21q11 deletion in three different cell lines. In two of the cell lines there was an additional supernumerary inv dup(22) or an r(22), respectively. In the third cell line the del(22) was the sole anomaly. FISH analysis showed that both the inv dup(22) and the r(22) included the DGS region. We hypothesize that an inter-chromosomal recombination between inverted repeats, together with a recombination between sister chromatids during meiosis I, gave rise to a deletion of 22q11 as well as an inv dup(22) containing the DGS region. The inv dup(22) was later rearranged into a ring chromosome during mitosis which was subsequently lost during cell division, thereby resulting in three different cell lines. This is the first case reported with an inv dup(22) and a del(22)(q11) in the same cell line. Our findings support a related mechanism in the formation of these two rearrangements mediated by low copy repeats. PMID- 11039584 TI - A reinvestigation of non-disjunction resulting in 47, XXY males of paternal origin. AB - We have used polymorphisms within the Xp/Yp pseudoautosomal region (PAR 1) to determine the frequency and location of recombination in 80 paternally derived 47, XXY males. Of 64 informative results, there were 10 single cross-overs, one double cross-over and 53 without a cross-over. Therefore 2/3 of 47, XXY males of paternal origin result from meiosis in which the X and Y chromosomes fail to recombine. This failure was not associated with the presence of an increase in recombination in the smaller Xq/Yq pseudoautosomal region (PAR 2) or with the presence of microdeletions within PAR 1. PMID- 11039585 TI - Fine mapping of a distinctive autosomal dominant vacuolar neuromyopathy using 11 novel microsatellite markers from chromosome band 19p13.3. AB - We previously mapped a distinctive autosomal dominant vacuolar neuromyopathy on human chromosome 19p13 in an 8cM region, delimited by D19S209 and D19S177 markers. We now report the fine mapping of the disease locus within an interval of 250 Kb by haplotype analysis performed using a set of 11 novel microsatellite markers isolated from the candidate region. PMID- 11039586 TI - Comparative study of the cardiovascular effects of losartan in normal and in water- and salt-depleted sheep. AB - The cardiovascular effects of losartan, a non-peptidic angiotensin II (ANG II) receptor antagonist, were studied in sheep. Eight normotensive, conscious sheep were tested twice: first under normal conditions and second when subjected to water and electrolytic depletion (furosemide 5 mg/kg twice a day for 3 days). Intravenous injection of 30 mg/kg losartan lowered the mean arterial blood pressure (MABP) in both control and water- and electrolyte-depleted sheep alike. The maximal decrease in MABP was significantly greater in diuretic-treated sheep than in controls (20.0 +/- 2.7 vs 9.3 +/- 1.1 mmHg) and occurred earlier (8.0 +/- 3.3 min vs 12.1 +/- 2.9 min). The decrease in blood pressure was associated with tachycardia in both controls and diuretic-treated sheep (+5.5 +/- 1.8 vs +11.3 +/ 3.9 beats/min). The vasopressor response to 0.1 microg/kg ANG II administered 30 min after losartan was completely antagonized. Two hours after losartan administration, MABP was on the increase in all animals and ANG II receptor blockade was partially obliterated in control sheep. The more marked cardiovascular effects recorded in diuretic-treated sheep as compared to control animals were associated with an increased activation of the renin-angiotensin system (plasma renin concentration: 6.51 +/- 1.33 vs 1.42 +/- 0.37 ng angiotensin I/ml/hr). PMID- 11039587 TI - Effects of lipid-related factors on adipocyte differentiation of bovine stromal vascular cells in primary culture. AB - The effects of several factors related to lipids on bovine adipocyte differentiation were investigated in primary culture. Adipocyte differentiation was assessed by development of glycerol-3-phosphate dehydrogenase (GPDH) activity and morphological observation. Addition of triglyceride mixture (Intralipid), caprylic acid and very low-, low- and high-density lipoproteins (VLDL, LDL and HDL) stimulated bovine preadipocyte differentiation in serum-free condition. Especially, VLDL strongly increased both cell protein contents and GPDH activity, suggesting that it stimulated both proliferation and differentiation of bovine preadipocytes. Under Intralipid-induced condition, differentiation of preadipocytes from subcutaneous adipose tissues was more evident than those from omental adipose tissues. However, such depot difference was not observed in medium supplemented with indomethacin, which is a peroxisome proliferator activated receptor (PPAR) gamma agonist. This suggests that the differentiation capacity of bovine preadipocytes was different between depots and such difference is dependent on the ability to utilize lipids as endogenous PPARgamma ligands. Therefore, lipid metabolites have the stimulatory effects on bovine adipocyte differentiation in vitro, and lipoproteins, especially VLDL, may play an important role in development of bovine adipose tissues in vivo. PMID- 11039588 TI - Comparative analysis of the putative amino acid sequences of chlamydial heat shock protein 60 and Escherichia coli GroEL. AB - The nucleotide sequences of the gene encoding chlamydial heat shock protein 60 (cHSP60) of 7 Chlamydia psittaci strains were determined. Comparison of sequences of the cHSP60 gene among chlamydiae showed high identities of the nucleotide sequences by 81.0% or greater and of the deduced amino acid sequences by 92.2% or greater. Comparison of the amino acid sequences between chlamydia and the other bacterial HSP60s resulted in the finding of three highly conserved regions, suggesting that these regions play a role in some function. In addition, 26- or 27-functional residues in the Escherichia coli GroEL out of the 28-residues are conserved in the amino acid sequences of the cHSP60. The data suggest that the function of the cHSP60 may be the same as that of the E. coli GroEL. PMID- 11039589 TI - Intestinal spirochetosis in wild sika deer (Cervus nippon yesoensis) infected with Brachyspira species. AB - Seven adult free-ranging sika deer (Cervus nippon yesoensis) were examined by histology, immunohistochemistry and electron microscopy for intestinal spirochetal infection. Histologically epithelial and goblet cell hyperplasia and edema of the lamina propria mucosa with macrophage and lymphocyte infiltration were observed in the cecum and colon in 6 of the 7 deer. Numerous argyrophilic spirochetes were present in the crypts and some had invaded epithelial and goblet cells and caused degeneration. Immunohistochemically the organisms stained positively with polyclonal antisera against Brachyspira (Serpulina) hyodysenteriae and B. pilosicoli. Ultrastructurally they were 6-14 microm long, 0.2-0.3 microm wide and had 4-6 coils and 13 axial filaments per cell; such features were closely similar to those in the Brachyspira species. These results showed that the spirochetes were capable of inducing enteritis in deer and this intestinal spirochete infection might already be prevalent among wild sika deer in Japan. There is a possibility that this spirochetal colitis is a new syndrome in sika deer and that the same and/or similar spirochetes have infected ruminants, including sika deer and cattle. PMID- 11039590 TI - Molecular cloning of equine chromogranin A and its expression in endocrine and exocrine tissues. AB - Chromogranin A (CGA) is a member of a family of highly acidic proteins co-stored and co-released with catecholamines in the adrenal medullary cells as well as in other neurons and paraneurons. The nucleotide sequence encoding equine CGA was determined using RT-PCR and rapid amplification of complementary DNA (cDNA) ends (RACE) techniques. A total 1,828 bp of the nucleotide sequence reveals that equine CGA is a 448-residue protein preceded by an 18-residue signal peptide. Comparison of the amino acid sequence of equine CGA with those of human, porcine, bovine, mouse, rat and frog CGA showed high conservation at the NH2-terminal 1-77 amino acids regions (94.8%, 93.5%, 92.2%, 81.8%, 83.1% and 66.2%, respectively) and COOH-terminal 314-430 amino acids regions (90.6%, 81.4%, 90.6%, 80.5%, 83.3% and 39.0%, respectively), as well as a potential dibasic cleavage site, whereas the middle portion showed marked sequence variation (52.5%, 49.1%, 38.9%, 26.6%, 27.9% and 6.2%, respectively). Northern blot analysis and RT-PCR elucidated the tissue distribution of equine CGA mRNA. Its expression was confirmed not only in the adrenal medullary cells but also in other organs (cerebrum, cerebellum, pituitary gland, spinal cord, liver, thyroid gland, striated muscle, lung, spleen, kidney, parotid gland and sublingual gland). Further, in adrenal chromaffin cells and pituitary cells of the anterior-intermediate lobe, the expression was confirmed by in situ hybridization with anti-sense CGA cRNA probe. PMID- 11039591 TI - Immunoprophylactic effect of chicken egg yolk immunoglobulin (Ig Y) against porcine epidemic diarrhea virus (PEDV) in piglets. AB - Porcine epidemic diarrhea virus (PEDV) is the causative agent of neonatal diarrhea in piglets, which causes high mortality rates. In this study, the immunoprophylactic effects of chicken egg yolk immunoglobulin (Ig Y) against PEDV were investigated in neonatal pigs. Ig Y was found to reduce the mortality in piglets after challenge exposures. The field application of Ig Y also revealed significant differences in survival rates of piglets given Ig Y, as compared with placebo or control. The results in this study indicated that Ig Y against PEDV could be an alternative way of supplementing prophylactic measures like colostral antibodies from sows. PMID- 11039592 TI - Attachment and penetration of canine herpesvirus 1 in non-permissive cells. AB - Canine herpesvirus 1 (CHV-1) has a relatively narrow host cell range when compared to other alphaherpesviruses. The early events of CHV-1 infection in a permissive Madin-Darby canine kidney (MDCK) and non-permissive cell lines. In order to quantify attachment and penetration, were investigated quantitative competitive PCR (QCPCR) method was established for quantitation of CHV-1 DNA. In all non-permissive cells tested, no significant decrease in viral attachment was observed. When CHV-1 was treated with heparin, viral attachment to MDCK cells was reduced by 25% of the input CHV-1 attached to MDCK cells even in the presence of 50 microg/ml heparin. However, the attachment of CHV-1 to non-permissive cells was severely impaired by heparin treatment. In permissive MDCK cells, about 80% of attached CHV-1 penetrated into cells. However, only 4-10% of CHV-1 attached to non-permissive cells penetrated into cells. Our data indicated that CHV-1, like other herpesviruses, attached to permissive MDCK cells through two mechanisms: the first one is through the interaction mediated by heparan sulfate (HS) on the cell surface and the second involves unidentified viral component and the cellular receptor. In contrast, the non-permissive cells lacked the cellular receptor for the second attachment mechanism and the defect in viral penetration into non-permissive cell might be related to the lack of the cellular receptor. PMID- 11039593 TI - Differential toxic effects of gentamicin on cultured renal epithelial cells (LLC PK1) on application to the brush border membrane or the basolateral membrane. AB - Aminoglycoside antibiotics are generally accepted to accumulate in renal proximal tubule cells from the luminal surface and show toxic effects on the cells. The binding affinity and membrane permeability of aminoglycoside antibiotics are different at the brush border membrane (BBM) and the basolateral membrane (BLM) of proximal tubule cells. This study was performed, therefore, to investigate the differential effects of the aminoglycoside antibiotic gentamicin (GM) on cultured LLC-PK1 cells, a pig kidney proximal epithelial cell line, after addition to the BBM or the BLM side. LLC-PK1 cells were cultured on microporous membranes until forming confluent monolayers, and then GM was added to either the BBM or the BLM side. GM caused release of enzymes from the organelles, with a higher level of release observed following addition to the BBM side than that to the BLM side. Patterns of [3H]GM uptake by the cells differed in a manner dependent on whether it was added to the BBM or the BLM side. That is, the cellular uptake from the BBM side increased with incubation time, while that from the BLM side showed rapid saturation. These results suggested that aminoglycoside antibiotics show differential effects on cultured proximal epithelial cells and have differential patterns of cellular uptake when added to the BBM or the BLM side. PMID- 11039594 TI - Roles of oxygen radical production and lipid peroxidation in the cytotoxicity of cephaloridine on cultured renal epithelial cells (LLC-PK1). AB - To clarify the mechanism of cephalosporin nephrotoxicity, the cytotoxic effects of cephaloridine (CER), a nephrotoxic cephalosporin antibiotic, on the pig kidney proximal tubular epithelial cell line (LLC-PK1) were studied in culture. CER increased the content of hydrogen peroxide and decreased the activity of catalase in the treated cells, followed by an increase in the content of lipid peroxide and decreases in both glutathione peroxidase activity and in the non-protein sulfhydryl content. The levels of NADPH-dependent hydrogen peroxide and superoxide anion production by microsomes prepared from LLC-PK1 cells, and by NADPH-cytochrome P-450 reductase purified from the rat renal cortex were significantly increased by paraquat. The production of these molecules was antagonized by p-chloromer-curibenzoate, an inhibitor of NADPH-cytochrome P-450 reductase. On the other hand, CER did not significantly affect the production of hydrogen peroxide or superoxide anions. These results suggested that the cytotoxic effect of CER on cultured LLC-PK1 cells was due to the increases in hydrogen peroxide and lipid peroxide levels and not microsomal oxygen radical production, and that the mechanism of this cytotoxicity is very different from that of paraquat which induces microsomal oxygen radical production. PMID- 11039595 TI - Comparison of response to immunotherapy by intradermal skin test and antigen specific IgE in canine atopy. AB - The intradermal skin test (IDST) and serologic allergy test (SAT) has been developed for confirming a diagnosis of canine atopy and determining allergens for immunotherapy. To determine the prevalence of causative allergens for canine atopic dermatitis in Japan, IDST and SAT were performed with the CMG Immunodot strips on 95 atopic dogs using 9 allergens. In addition, we compared agreement rate, sensitivity and specificity between them (using IDST as the standard). The allergen most commonly positive in both tests was house dust mites (IDST: 69.5%, SAT: 48.4%). Moreover, Japanese cedar, mugwort and grass mix were detected as attendant causative allergens. Agreement rates between the two tests ranged from 67.4% to 96.8%; the overall mean agreement rate were 81%. SAT was shown to have sensitivity to IDST ranging from 16.7 to 68.2%. The specificities were very high for all allergens, on the order of 94.9-100% (median=98.7%). Finally, the efficacy of immunotherapy was evaluated on 27 atopic dogs based on IDST (15 dogs) and SAT (12 dogs) results. Overall, 60% (9/15) of the IDST group and 66.8% (8/12) of the SAT group experienced a 50% to 100% reduction in their symptomatology. No significant differences were found in response to immunotherapy during the follow up period between allergen selection methods. These results indicate the value of serologic tests as an aid to identifying an allergen solution for immunotherapy. PMID- 11039596 TI - Pathogenicity and gene analysis of adenovirus from pigeons with inclusion body hepatitis. AB - The pathogenicity of an adenovirus isolated from pigeons (Pigeon adenovirus, PA) with inclusion body hepatitis in Taiwan was investigated in specific-pathogen free (SPF) chicks and in racing pigeons. One-day-old SPF chicks were inoculated subcutaneously with 10(4) 50% tissue culture infective dose (TCID50, low dose) and 10(8) TCID50 (high dose) of the virus, respectively. The chicks began to die three days post inoculation (DPI) with high dose of the virus and the mortality reached 100%; the chicks began to die 6 DPI and the mortality reached 90% at 14 DPI with low dose. The adult pigeons seemed to be resistant to the PA. However, this virus decreased the production of antibody against Newcastle disease virus in pigeons. It is found that this PA belongs to genetic group D from the restriction patterns produced by BamH I and Hind III. PMID- 11039597 TI - In vivo and in vitro tests showing sensitization to Japanese cedar (Cryptomeria japonica) pollen allergen in atopic dogs. AB - Using both in vivo and in vitro tests, dogs with atopic dermatitis were examined for sensitization with Japanese cedar (Cryptomeria japonica, CJ) pollen allergen. Ten dogs with clinical manifestation of atopic dermatitis were shown to be sensitized to CJ pollen based on the results of intradermal skin test and serum antigen-specific IgE test. In vitro lymphocyte stimulation test showed blastogenic response after stimulation with crude antigen of CJ pollen in all of the 5 cases examined. The peripheral leukocytes showed increased histamine release after stimulation with crude antigen of CJ pollen in 2 cases examined. These data indicate that a proportion of dogs with atopic dermatitis is sensitized to CJ pollen in a cell-mediated manner and show immediate phase reaction of type I hypersensitivity. PMID- 11039598 TI - Distribution of immunoglobulin isotypes and subisotypes in equine guttural pouch (auditory tube diverticulum). AB - To clarify the functions of the equine guttural pouch, the distribution of various immunoglobulin isotypes and subisotypes in the guttural pouch mucosa were examined in healthy horses. IgGa was present in the mucosa of guttural pouch, mucosal lymph nodules and submucosal lymph nodules. IgM was scattered in the mucosal lymph nodules and in the germinal centers of the submucosal lymph nodules. IgGc was recognized only in the submucosal lymph nodules. These immunoglobulin isotypes and subisotypes were found in lymphocytes and plasma cells. On the other hand, IgA was detected in glandular epithelial cells and the surface layer of the mucosal epithelium, as well as in free cells. This finding suggests that IgA is secreted through the glandular epithelium. Based on the above findings, we conclude that the guttural pouch has phylactic ability. PMID- 11039599 TI - Spinal projections of cat primary afferent fibers innervating caudal facet joints. AB - The spinal projections of afferent fibers innervating the facet joints between caudal vertebrae were examined by the use of anterograde transport of wheat germ agglutinin-horseradish peroxidase (WGA-HRP). Experiments were performed on 5 adult cats in which spinal dorsal roots below the 2nd sacral segment (S2) on the right side were cut. Injections of WGA-HRP into the caudal facet joints gave rise to extensive cranio-caudal distribution of WGA-HRP positive products along the spinal cord, indicating that many afferent fibers innervating unilateral facet joints terminate bilaterally in laminae I-II, V-VI and X of the thoracic, lumbar, sacral and caudal spinal cord. These afferent fibers may convey a series of sensory information from the caudal facet joints to the spinal cord. PMID- 11039600 TI - Mast cell tumor in the nasal cavity of a dog. AB - An 11-year-old male Shetland sheepdog displayed epistaxis and nasal discharge from the left nasal foramen. Cytological examination of a smear sample obtained by rhinotomy revealed neoplastic mast cells in the nasal cavity, a definitive diagnostic sign of mast cell tumor. The case was treated by surgery combined with radiotherapy and chemotherapy. Eighteen days after the last treatment, marked enlargement of the mandibular lymph nodes and facial edema developed, and the dog was euthanized at the owner's request. At necropsy, metastatic proliferation of mast cells was confirmed in the lymph nodes and liver, but no neoplastic mast cells were observed in the nasal cavity. PMID- 11039601 TI - Cloning of canine cDNA encoding tektin. AB - Tektins are a group of proteins that form filamentous polymers in the walls of ciliary microtubules. The cloning of canine cDNA encoding tektin, was carried out and identified from the testis of beagle dog. Canine tektin cDNA is 1,523 bp in length, has an open reading frame of 1,281 bp nucleotides encoding a protein of 426 deduced amino acids. The predicted amino acid sequence has 77% and 33-50% of homology with the murine tektin and the sea urchin tektins. The amino acid sequence RPNVELCRD and four cysteine residues were conserved in the dog, mouse and sea urchin, suggesting the functional significance of this protein domain and the amino acid residues in the tektin proteins. PMID- 11039602 TI - Anesthetic management with sevoflurane and oxygen for orthopedic surgeries in racehorses. AB - Eighty-five thoroughbred racehorses with various types of fracture were subjected to arthroscopic surgery (44 horses) or internal fixation (41 horses) under sevoflurane anesthesia. The mean end-tidal sevoflurane concentration during anesthesia ranged from 2.5 to 2.8%. PaCO2 was maintained between 50 and 65 mmHg by controlled ventilation. The mean arterial blood pressure was maintained above 65 mmHg by infusion of dobutamine and fluids, however, heart rate significantly increased with time. Recovery from anesthesia was calm and smooth in almost all cases. No apparent complication was observed during and after anesthesia in all cases. Therefore, sevoflurane anesthesia is considered to be safe and useful for orthopedic surgery in racehorses. PMID- 11039603 TI - Ultrastructural study of Langerhans cells in equine insect hypersensitivity "Kasen". AB - Ultrastructural features of Langerhans cells (LCs) of equine "Kasen" were studied. Electron microscopic observation revealed that LCs were dendritic and had irregular nuclear membranes. A number of Birbeck granules (Bgs) of various types were observed in the cytoplasm of LCs. In LCs in the upper stratum spinous, many Bgs were observed (Type 2 LC). LCs in the epidermo-dermal junction (EDJ) had a few Bgs, vesicles (multivesicular bodies) and highly electron-dense granular endosomes in the cytoplasm (Type 3 LC). Inactive LCs were also observed between the keratinocytes (Type 1 LC). Various types of LCs observed in the skin lesions of equine "Kasen" were interpreted as representing those that recognize, intake and process antigens. PMID- 11039604 TI - Basic and update knowledge of intervertebral disc herniation: review. AB - Lumbar disc herniation is one of the most common causes of low back pain and/or sciatica. However, the pathogenesis of lumbar disc herniation, low back pain, and sciatica has not been fully understood. Inflammation in nerve root and dorsal root ganglia induced by nucleus pulposus may play an important role in the pathogenesis of spinal pain. I reviewed the basic and update papers regarding lumbar disc herniation. Herniated nucleus pulposus had been considered an enchondroma occurred from intervertebral disc, historically. At present, however, it is emphasized that nucleus pulposus has an inflammatogenic properties to affect the nerve root function, structure, vascular permeability, and pain. PMID- 11039605 TI - VEGF-dependent signaling in retinal microvascular endothelial cells. AB - We examined the effect of vascular endothelial growth factor (VEGF) on intracellular signal transduction pathways using isolated bovine microvascular endothelial cells (BREC). When cell growth was determined by [3H]thymidine incorporation, it was significantly stimulated by VEGF stimulation. In situ hybridization results also demonstrated that c-fos expression was enhanced by the stimulation. Although BREC expressed Flt-1 and Flk-1 as VEGF receptors at similar levels, VEGF stimulation preferentially enhanced the activity of Flt-1 tyrosine kinase. This stimulation initiated an increase in the level of GTP-form Ras and the activation of mitogen activated protein kinase (MAPK). On the other hand, BREC expressed the Janus kinase (Jak) family members Jak1, Jak2, and Tyk2, and the signal transducers and activators of transcription (Stat) family members Stat1, Stat3, and Stat6. These molecules were tyrosine phosphorylated under culture conditions used, and the phosphorylation of Tyk2 and Stat6 was specifically enhanced by VEGF stimulation. These results demonstrate that, in addition to Ras/MAPK pathways, the Flt-1/Tyk2/Stat6 pathway is important in VEGF signaling in BREC. These signal transduction systems may regulate the growth of retinal endothelial cells. PMID- 11039607 TI - Re-assessment of stage I uterine cervical carcinoma according to revised JSGO (1997) staging. AB - BACKGROUND: The Japanese Society of Gynecologist and Obstetrician (JSGO) revised criteria for early uterine cervical cancer. OBJECTIVE: In this study, we investigated the value of the revised JSGO criteria. METHOD: Retrospective review was performed for 70 patients with tumors limited to the uterine cervix who were classified stage I under old JSGO criteria. RESULT: Forty patients were re classified into stage IA1 and 4 patients into stage IA2. Incidence of lymph vascular infiltration (LVI) in stage IA1 and IA2 were 5% and 25%. There was one patient with stage IA1 disease who had nodal metastasis and no patients with stage IA2 disease. There were 14 patients with stage IB1 disease and 12 patients with stage IB2 disease. Mortality of patients with stage IB2 disease was 25% and significantly higher than that of patients with stage IA and IB1 disease (P<0.001 and <0.05) CONCLUSION: Although the revised JSGO criteria for early cervical cancer are acceptable for assessment of patients, the therapy of patients is still controversial. We recommend that patients with stage IA1 and negative LVI should have less radical hysterectomy and patients with stage IA1 with LVI or stage IA2, IB should have radical hysterectomy. PMID- 11039606 TI - Evaluation of perioperative administration of methylprednisolone sodium succinate and urinary trypsin inhibitor for prevention of surgical stress. AB - The authors could confirm that the laparoscopy-assisted cholecystectomy (LAC) elicited less postoperative biological responses compared to the ordinary cholecystectomy under laparotomy (OCL), when granulocyte elastase (GE)-alpha1 protease inhibitor complex (GEcomplex), interleukin-6, pancreatic secretory trypsin inhibitor (PSTI) and alpha1-antitrypsin (alpha1-AT) were used as biological response markers. Perioperative administrations of methylprednisolone sodium succinate (MPSL: 10 mg/kg body weigh) or MPSL with urinary trypsin inhibitor (UTI) could suppress such postoperative reactions after OCL down to the levels after LAC, especially immediately after surgery. Preoperative MPSL followed by continuous infusion of UTI for 3 days exerted the most prominent suppressive effects on these markers compared to the effect of the preoperative MPSL alone as well as the preoperative administration of MPSL followed by UTI infusion for only one hour. Bolus administration of MPSL induced no lymphocytopenia. Decreased plasma level of alpha1-AT immediately after operation is thought to be due to consumption in binding to GE as well as other lysosomal enzymes, while production of rapid turn over proteins are still not accelerated in the liver. In early postoperative phase after administration of MPSL, administration of UTI was efficacious to prevent fluctuation of biological response markers. Clinical applications of these drugs might be approved especially for those patients with poor risk. PMID- 11039608 TI - The effect of detergents on chemical occult blood test. AB - OBJECTIVES: Our previous investigation showed that toilet sanitizers and their main constituents, detergents, caused false negative reactions of the immunological occult blood test. Therefore, we investigated the effects of detergents on the chemical occult blood test. MATERIALS AND METHODS: Twenty-nine kinds of detergents were added to a Hb solution and measured Hb concentration by a quantitative chemical occult blood test (modified colorimetric o tolidine method) and the results were compared with those by the immunological test. The detergents' effects on Hb spectra were also analyzed. RESULTS: The chemical test was affected slightly by most of the detergents tested although the immunological test was markedly to moderately affected. The spectrum of hemoglobin was not affected by the detergents examined except for the high concentrations of SDS and DTAB. CONCLUSION: The chemical occult blood test is affected by some detergents, but less than the immunological tests. PMID- 11039609 TI - Rhabdomyolysis and aggravation of arthritis in a rheumatoid arthritis patient as a result of sepsis due to Staphylococcus aureus infection of a rheumatoid nodule; a catastrophic outcome. AB - A 63-year-old man with rheumatoid arthritis presented with rhabdomyolysis and intractable arthritis of acute onset. He was diagnosed to have sepsis due to Staphylococcus aureus infection through of an ulcerated rheumatoid nodule. Staphylococcus aureus isolated from pus in the ulcerated rheumatoid nodule and a blood sample obtained from the heart post-mortem produced the toxic shock syndrome toxin-1 (TSST-1). The TSST-1 and/or unmethylated CpG motifs in the oligonucleotides present in a bacterium, Staphylococcus aureus in this case, might be implicated in the induction of rhabdomyolysis and intractable arthritis. PMID- 11039610 TI - Contemporary cardiac imaging: an overview. AB - Comprehensive cardiac assessment embraces virtually every imaging modality and includes information about coronary vascular anatomy as well as cardiac morphology, function, perfusion, metabolism, and tissue characterization. Through sophisticated computer processing and image analysis, newer imaging technologies such as computed tomography (CT), magnetic resonance (MR), MR spectroscopy, and positron emission tomography now provide quantitative information that may obviate more invasive angiographic assessment. Currently, no single imaging technology realizes all questions relating to cardiac form and function, and many of the technologies overlap in the content and quality of information they provide. This overview seeks to provide a broad perspective on current cardiac imaging, articulating the benefits of various technologies and their limitations. PMID- 11039611 TI - The cardiac silhouette. AB - The cardiac silhouette as seen on a routine chest film contains considerable information regarding the presence of heart disease, the nature and severity of the disease, and its prognosis. The changes that are seen directly reflect the altered anatomy and physiology of the heart. Appreciation of these factors allows for a reasonable, logical approach to interpretation of the cardiac silhouette. PMID- 11039612 TI - Assessment of cardiac function: magnetic resonance and computed tomography. AB - A complete cardiac study requires both anatomic and physiologic evaluation. Cardiac function can be evaluated noninvasively by magnetic resonance imaging (MRI)or ultrafast computed tomography (CT). MRI allows for evaluation of cardiac function by cine gradient echo imaging of the ventricles and flow analysis across cardiac valves and the great vessels. Cine gradient echo imaging is useful for evaluation of cardiac wall motion, ventricular volumes and ventricular mass. Flow analysis allows for measurement of velocity and flow during the cardiac cycle that reflects cardiac function. Ultrafast CT allows for measurement of cardiac indices similar to that provided by gradient echo imaging of the ventricles. PMID- 11039613 TI - Computed tomography and magnetic resonance imaging of patients with valvular heart disease. AB - Although computed tomographic (CT) and magnetic resonance (MR) evaluation of patients with valvular heart disease is almost never performed as a first line of diagnostic intervention, their performance does provide important morphologic and physiologic information concerning the etiology and the current status of the valvular dysfunction. Evaluation of chamber and great artery size as well as ventricular wall thickness provide the basis for diagnosing and analyzing severity of valvular heart disease. Furthermore, additional findings, including calcification and evidence of interstitial pulmonary edema, increase diagnostic sensitivity and confidence in diagnosis. MR examination has the advantage over CT of providing direct demonstration of the signal void jets of dysfunctional valves, as well as a means of quantitating regional and global ventricular function and severity of valvular pressure gradients. PMID- 11039614 TI - Imaging of cardiac and paracardiac masses. AB - Magnetic resonance imaging (MRI) and computed tomography (CT) are important imaging modalities for the noninvasive characterization of cardiac and paracardiac masses. They are, in general, superior to other modalities (e.g., echocardiography) in their ability to delineate the exact location and the extent of the lesion and to demonstrate the effects of the lesion on surrounding structures. MRI and CT may also be helpful in suggesting a specific diagnosis, because some tumors have rather characteristic locations and appearances. In addition, both modalities can be extremely helpful in both treatment planning and posttreatment follow-up because they are noninvasive, reproducible, and enable detection of residual or recurrent mass. PMID- 11039615 TI - Imaging of pediatric congenital heart disease. AB - The evaluation of a patient with a heart murmur and congenital heart disease is diagnostically very challenging. Multiple advanced techniques aid in the diagnosis of simple and complex malformations. Interventional procedures and surgical corrections have allowed most patients to enjoy a good and productive lifestyle. PMID- 11039616 TI - Lecturing on the Web. PMID- 11039617 TI - Airways obstruction in patients with sarcoidosis: expiratory CT scan findings. AB - In patients with pulmonary sarcoidosis, air trapping as evidenced by expiratory high-resolution computed tomography (HRCT) is not specific for a given stage of disease. Air trapping can occur at the level of the secondary lobule, as well as in distributions suggesting sublobular, subsegmental, and segmental involvement. While air trapping can be a nonspecific finding, it is a common feature in patients with pulmonary sarcoidosis and is a supportive diagnostic finding. PMID- 11039618 TI - Pulmonary artery and aortic sarcomas: cross-sectional imaging. AB - Sarcomas of the major arteries are rare tumors often misdiagnosed due to nonspecific symptomatology. The authors present three cases of pulmonary artery and aortic sarcomas that were initially believed to be more common diseases. Modern imaging techniques including helical computed tomography (CT) and magnetic resonance imaging (MRI) are increasing the frequency of premortem diagnosis of these entities and aided surgical planning. PMID- 11039619 TI - Intrapleural rupture of a cystic thymoma. AB - Although cystic degeneration of a thymoma is not uncommon, rupture of a cystic thymoma is rare. The authors report a patient with sudden chest pain and dyspnea due to rupture of a cystic thymoma into the right pleural space. PMID- 11039620 TI - Kaposi sarcoma in a renal transplant patient. AB - The radiographic abnormalities of primary Kaposi's sarcoma of the lung in a patient with a renal transplant are reported. The findings are similar to other malignancies and infections that are well recognized in the renal transplant population. In the appropriate clinical setting, the radiologist should consider the diagnosis of Kaposi's sarcoma even in the absence of cutaneous lesions as reducing immunosuppression can be curative therapy. PMID- 11039621 TI - Intrathoracic migration of Steinmann pins. AB - The migration of surgical wires and pins placed for repair of orthopedic injury is well recognized. Such migration usually follows a retrograde path and the wires protrude near their entry point into the native bone. Occasionally, the migration occurs in an antegrade manner and produces injury. We describe a case where three Steinmann pins placed for fixation of a humeral neck fracture migrated, one slipping backwards towards the humeral entry point and two pins migrating into the thorax. PMID- 11039622 TI - Developing clinical guidelines in dentistry--what are the opportunities for international collaboration? PMID- 11039623 TI - Oral health related quality of life--views of the public in the United Kingdom. AB - OBJECTIVE: This study was designed to determine the United Kingdom public's perception of how oral health affects quality of life (QoL) and to determine socio-demographic variations in these perceptions. RESEARCH DESIGN: The vehicle for this study was the ONS Omnibus Survey in the UK. A random probability sample of 2,668 eligible addresses was selected from the British Postcode Address File. Setting The data were collected by qualitative, face-to-face interviews with respondents, nation-wide, in their homes, about how their oral health status affected their QoL. PARTICIPANTS: 1,778 adults aged 16 years or older across the UK took part in the study. RESULTS: 75% (1,340) believed their oral health either enhanced or reduced their QoL. Most frequently, this was perceived as being the result of its effect on eating. comfort and appearance. Other ways in which QoL was affected are also presented. Sociodemographic variations were apparent. For example, people from higher socio-economic backgrounds believed that their oral health enhanced their QoL to a greater degree (OR=1.46, CI=1.20, 1.78) than the lower socio-economic groups. Women claimed that their oral health had a greater negative effect on QoL than did men (OR=1.36, CI=1.11, 1.64). Younger people (16 64 years old) reported that their oral health status reduced and enhanced QoL more than older adults (65 years and over) (OR=1.59, CI=1.23, 2.04). CONCLUSIONS: The study shows that the UK public perceives oral health as affecting their QoL in a variety of physical, social and psychological ways and that significant socio-demographic variations exist in these perceptions. PMID- 11039625 TI - Optimum bitewing examination recall intervals assessed by computer simulation. AB - OBJECTIVE: To assess the effect of altering bitewing examination recall intervals on health gain from dental restorative treatment and to determine optimum recall intervals under varying clinical conditions. DESIGN: A computer simulation of the caries process in posterior approximal tooth surfaces. The effect of superimposing restorative treatment, based on diagnoses from bitewing radiological examinations carried out at differing time intervals, was incorporated. Input data included caries attack rates, median survival times of restorations, and sensitivity (Sn) and specificity (Sp) of treatment decision making by a high (A) and a low (B) performing dentist. PARTICIPANTS: A hypothetical population, initially 14-15 years old. INTERVENTIONS: Class II amalgam restorations. OUTCOME: Health gain in utility based units (UBUs) was assessed relative to interim end point UBUs pertaining under 'do nothing scenarios'. RESULTS: One thousand approximal surfaces, designated initially as 920 sound, 51 carious and 29 filled were followed in the model over 10 years. The greatest health gain (39.33 UBUs) was from dentist A (Sn = 0.23, Sp = 0.99, restoration median survival time = 20 years, caries rate = 4.4% per annum, optimal recall interval between bitewing radiological examinations = 7 months). The least was from dentist B (Sn = 0.52, Sp = 0.88, median survival time = 5 years. caries rate = 0.0% per annum, optimal recall interval between bitewing radiological examinations > 120 months) representing a loss of 16.79 UBUs compared with 'do nothing'. CONCLUSIONS: In the best interests of their patients, it would seem that dentists need to exercise considerable caution in making positive decisions to restore approximal tooth surfaces on the basis of bitewing radiographic evidence and that for some dentists current guidelines for bitewing examination intervals would appear to be too permissive. PMID- 11039624 TI - Preschool children's consumption of drinks: implications for dental health. AB - OBJECTIVE: To examine drinking patterns in pre-school children and their relationship to percentage of energy intake from non-milk extrinsic sugars. DESIGN: Secondary analysis of data from the national diet and nutrition survey (NDNS) relating to the dietary intakes of a representative sample of pre-school children in the UK. SUBJECTS: 1,675 children aged 1.5 to 4.5 years surveyed between July 1992 and June 1993. OUTCOME MEASURES: Proportion of consumers, average daily frequency of consumption and estimated seven day volume of consumption of different drinks. Percentage of average daily energy intake obtained from non-milk extrinsic sugars (NMES). RESULTS: Soft drinks were the most commonly consumed drinks followed by whole milk and diet or low sugar varieties of soft drinks. Half the sample were estimated to consume more than 1.5 litres of soft drinks and whole milk and over a litre of diet or low-sugar soft drinks per seven days. Fifty-six per cent of the children consumed soft drinks more than once a day. The youngest children (1.5-2.5 years) were more likely to consume whole milk and less likely to consume diet, soft drinks and skimmed milk than other age groups. Children from manual home backgrounds consumed more tea and coffee and were less likely to consume fruit juice than those from non-manual backgrounds. Drinks contributed 23% to total energy intake and 39% of NMES intake. Consumption of soft drinks, fruit juice and whole and semi-skimmed milk accounted for 59% of variance in percentage of energy from NMES. CONCLUSIONS: A large proportion of pre-school children consume considerable quantifies of soft drinks which have little or no nutritional value and are high in cariogenic non milk extrinsic sugars. This has implications for children's dental and general health. Recommendations for drinks consumption should be included in food policy guidelines for pre-school children. PMID- 11039626 TI - Preventive service utilisation as a predictor for emergency dental examinations. AB - OBJECTIVE: To quantify the relationship between receipt of routine dental care and the use of non-trauma related emergency dental services. DESIGN: A multiple logistic regression was run on administrative dental claim and encounter data. The model dependent variable was the use of non-trauma related emergency dental care. Predictors included previous year oral examinations, radiographs, dental cleanings and, as a control, member age. SETTING: Administrative data were obtained from a dental health maintenance organisation located in the state of Texas. SUBJECTS: Claim and encounter data for 2,947 insured members were used, representing experience from 1995 through 1996. OUTCOME MEASURES: The outcome of interest was the use of non-trauma related emergency dental services. RESULTS: Results demonstrated empirically that those who availed themselves of preventive dental services were significantly less likely to use non-trauma related emergency services (P<0.01). The probability of needing non-trauma related dental services in 1996 was 42.7% lower among those who had an examination in 1995 when compared with those who did not. When analysed in a simple logistic regression, dental cleanings in 1995 were also significantly associated with a decreased probability of needing non-trauma related emergency services. However, this relationship did not hold in the controlled model, which was probably due to multicollinearity. CONCLUSIONS: This study provides evidence of the value of periodic preventive dental examinations and services. Those who receive such services are less likely to use non-trauma related emergency dental services. PMID- 11039627 TI - A survey of Scottish primary care dental practitioners' oral cancer-related practices and training requirements. AB - OBJECTIVE: The study sought to investigate current examination habits and preventive practices of Scottish dental primary care professionals, with respect to oral cancer, and to determine any training needs of these practitioners in relation to the disease. BASIC RESEARCH DESIGN: A questionnaire was sent to a random sample of 331 general dental practitioners and community dental officers throughout Scotland, achieving an overall response rate of 68%. MAIN OUTCOME MEASURES: The study investigated examination and health promotion practices in relation to oral cancer. Confidence in, and barriers to, participating in these activities were studied, and information sought both on past training and future education needs regarding oral cancer. RESULTS: Although 58% of respondents reported examining regularly for signs of oral cancer in those aged >16 years, 63% indicated they felt less than confident in detecting oral cancer, with only 43% expressing confidence about discussing suspicious findings with patients. Practitioners were well aware of the importance of smoking and alcohol as risk factors, but had mixed views on the health-promoting role of the dentist regarding these issues. Furthermore, while only 3% reported training on these topics, over half expressed a desire to develop appropriate counselling skills. Overall, 87% and 79% of respondents wanted further training in oral cancer detection and prevention, respectively. CONCLUSION: The study indicated a need for continuing education programmes for dental primary care practitioners in oral cancer-related activities. Postgraduate education, utilising a variety of media formats, should aim to improve diagnostic skills and seek to increase practitioners' participation in both smoking and alcohol counselling. PMID- 11039628 TI - Trends in self-reported use of dental services among Finnish adults during two decades. AB - OBJECTIVE: To analyse trends in the prevalence of and determinants for dental care utilisation among Finnish adults of working age during two decades. BASIC RESEARCH DESIGN AND PARTICIPANTS: The National Public Health Institute has, since 1978, conducted annual surveys of health behaviour among the whole Finnish adult population (aged 15-64 years). Data collected through mailed questionnaires have included questions on dental utilisation. Sample sizes have varied from 5,000 to 6,000 and response rates from 68% to 85%. RESULTS: The use of dental services increased in the period 1978 to 1997 from 53% to 64% (P<0.001) of the population. Women and persons belonging to the younger age groups were the most frequent visitors, and older persons the least frequent due to the greater prevalence of edentulousness. During the period, differences in the use of services associated with region, level of education and occupation declined. During the first decade, a clear increase in dental utilisation was found in younger age groups and during the latter decade in older age groups. Significant predictors for the utilisation of services were the number of missing teeth, age, gender, occupation and toothbrushing frequency in 1997. CONCLUSIONS: The utilisation of dental services has increased slowly since 1978 but remains lower than in other Nordic countries. In a country where the supply of services is abundant, the major determinant of the use of services is the number of teeth an individual has. PMID- 11039629 TI - Oral health status of 12-year-old schoolchildren in the province of Kadiogo, Burkina Faso. AB - OBJECTIVE: To assess the oral health status of 12-year-old schoolchildren in Kadiogo province, Burkina Faso. DESIGN: A cross-sectional epidemiological study in which a trained dentist performed clinical examinations. SETTING: Twenty primary schools in Kadiogo Province. SUBJECTS: Three hundred randomly selected children. OUTCOME MEASURES: Oral health status was recorded according to WHO criteria. Demographic characteristics (gender, type of school and geographic location) were also recorded. RESULTS: The results showed that 50% of the subjects examined were caries-free. The mean DMFT was 1.72+/-0.12 with a significant difference between urban and rural areas (P<0.05). Decayed teeth were untreated in this population, only 7% of whom had a healthy periodontium. Statistically significant differences were observed according to gender (P<0.02) and area (P<0.04). In the whole sample, 81% presented no malocclusion and only two questionable cases of fluorosis were observed. The rate of need for periodontal treatment was 83 to 100% in rural areas; 21% of the children needed at least one extraction and approximately 50% required conservative care. CONCLUSIONS: The mean DMFT found in this study placed the Kadiogo province in the low DMFT category defined by WHO and within the global goal of Health for All by the Year 2000 at 12 years old. However, decayed teeth were untreated, and periodontal status was very poor. There is therefore a need for appropriate preventive measures. PMID- 11039630 TI - Dental caries and treatment experience of adults from minority ethnic communities living in the South Thames Region, UK. AB - OBJECTIVE: To describe the dental caries and treatment experience of groups of adults from minority ethnic communities living in the South Thames Region of England. DESIGN: Cross-sectional clinical study. SETTINGS: Community, religious and educational centres for adults from minority ethnic communities. PARTICIPANTS: Snowball sample of 928 adults from 44 ethnic groups including: Black Caribbean (141), Black African (134); Pakistani (123); Indian (190); Bangladeshi (78); Chinese/Vietnamese (143) and 119 from other groups. RESULTS: More participants were dentate or had 18 or more sound and untreated teeth than adults living in the same area (Todd and Lader, 1991). Duration of residence in the United Kingdom predicted caries or treatment experience in the sample as a whole and in Chinese/Vietnamese people. Increased DMFT was predicted by age and by history of visiting a UK dentist in the sample as a whole and in the Black African group. CONCLUSIONS: Effect of duration of UK residence on presence and extent of caries suggests that oral health may be better among adults from these ethnic minority groups than among the general population. However, the differences can also be attributed to sampling bias and old comparison data. Better sampling strategies are required for research of this type. PMID- 11039631 TI - The dental caries experience of 14-year-old children in the United Kingdom. Surveys coordinated by the British Association for the Study of Community Dentistry in 1998/99. AB - DESIGN: This paper reports the results of standardised clinical caries examinations of 121.550 14-year-old children from across the United Kingdom, Jersey and the Isle of Man. These 1998/99 coordinated surveys are the latest in a series which seeks to monitor the dental health of children and to assess the delivery of dental services. METHOD: The criteria and conventions of the British Association for the Study of Community Dentistry were used. Representative samples were drawn from participating health authorities and boards and caries was diagnosed at the caries into dentine (D3) threshold using a visual method without radiography or fibreoptic transillumination. RESULTS: These demonstrated, once again, a wide variation in prevalence across the United Kingdom, with mean values for D3MFT for the current English regional offices (of the National Health Service) and the other UK countries ranging from 1.17 in West Midlands to 3.65 in Northern Ireland. The mean value for D3MFT across the United Kingdom was 1.76 (D3T=0.59, MT=0.15, FT=1.02). Overall, 54% of children had evidence of caries experience at the dentinal level (D3MFT>0), although the means ranged between 43% (South East) and 78% (Northern Ireland). The mean D3MFT for those with disease at this threshold was 3.24. Trends over time demonstrate an improvement of 10% in overall D3MFT for Great Britain since 1994/95, compared to the 21% seen over the previous four year period. Over recent years the overall trend in this age group seems to be towards lower values. However, there has been no improvement in mean MT since 1994/95, while FT and care index have fallen. The number of fillings provided in 1998/99 and thus the care index, remains low, on average across the UK, only 58% of the dentinal caries experience identified by survey examinations of permanent teeth was seen as fillings (range in individual districts and boards: 34% to 83%). CONCLUSION: Taken together, these findings demonstrate the continuing need for more effective preventive strategies and treatment services for this important age group. PMID- 11039632 TI - Re: Changing trends in South Wales flouride prescription dispensing (1993-7) [Community Dental Health (1999) 16, 145-148]. PMID- 11039633 TI - Re: Changing trends in South Wales flouride prescription dispensing (1993-7) [Community Dental Health (1999) 16, 145-148]. PMID- 11039634 TI - Re: Caries index distributions and inequality. PMID- 11039635 TI - Case report: reduction of low back pain in a professional golfer. AB - Previous research agrees that the majority of injuries that affect male golfers are located in the lower back and that they are related to improper swing mechanics and/or the repetitive nature of the swing. This study describes the trunk motion and paraspinal muscle activity during the swing of a golfer with related low back pain (LBP) and assesses the effect of a 3-month period of muscle conditioning and coaching on these variables. Motion of the trunk was measured using three-dimensional video analysis and electromyograms (EMGs) were recorded from the same six sites of the erector spinae at the start and end of the 3-month period. At the end of the period, the golfer was able to play and practice without LBP. Coaching resulted in an increase in the range of hip turn and a decrease in the amount of shoulder turn, which occurred during the swing. In addition, a reduction in the amount of trunk flexion/lateral flexion during the downswing occurred in conjunction with less activity in the left erector spinae. These changes may serve to reduce the torsional and compressive loads acting on the thoracic and lumbar spine, which in turn may have contributed to the cessation of the LBP and would reduce the risk of reoccurrence in the future. In conclusion, further research with more subjects would now be warranted in order to test the findings of this program for the prevention of low back in golfers as piloted in this case report. PMID- 11039636 TI - Cardiovascular load of competitive golf in cardiac patients and healthy controls. AB - PURPOSE: Sports in cardiovascular patients (CVP) should serve for risk factor management, increase of exercise capacity, and reintegration into daily life. Competition of cardiac patients with healthy sportsmen is often discouraged and thus reintegration hampered. Golf, with its endurance component and exceptional rules (e.g., the handicap) should be an alternative. METHODS: In 20 male golfers (65.2 +/- 6.1 yr, 1.4 +/- 0.3 W x kg(-1) body weight (approximately 4.8 METs)) with cardiovascular diseases and eight controls (C) (62 +/- 5 yr, 2 +/- 0.4 W x kg(-1) body weight (approximately 6.9 METs)), the performance assessed in the laboratory (ergospirometry, serum lactate) allowed for comparison of the cardiovascular load on the golf course (lactate, Holter monitoring, blood pressure, urine catecholamines). RESULTS: In comparison with in the hospital, resting heart rates were significantly (P < 0.001) elevated in both groups immediately before the tournament (CVP: 76.1 +/- 10.8 vs 90.1 +/- 8.6 bpm; C: 74.8 +/- 6.3 vs 92.3 +/- 9.7 bpm). On the course, the mean heart rates of the patients were closer (P < 0.01) to the anaerobic threshold (105.4 +/- 11.0 vs 115.3 +/- 10.8 bpm) in comparison with controls (100.5 +/- 7.3 vs 125.6 +/- 16.6 bpm) corresponding to 0.9 +/- 0.3 W x kg(-1) (approximately 3.1 METs) or 76.0 +/- 13.1%VO2max (CVP) and to 0.9 +/- 0.2 W x kg(-1) (approximately 3.1 METs) or 55.3 +/- 9.1%VO2max (C). Serum lactate levels were 1.36 +/- 0.7 mmol x L(-1) (approximately 12.4 +/- 6.4 mg x dL(-1)) (CVP) and 1.1 +/- 0.4 mmol x L(-1) (approximately 9.1 +/- 3.6 mg x dL(-1)) (C). In patients, arrhythmias were lower in quantity and quality (LOWN) in comparison with other activities as registered by means of the 24-Holter-ECG. CONCLUSION: In cardiovascular patients, competitive golf reaches an intensity that may positively influence cardiovascular risk factors, depending on the type of the course and may provide patients the desired integration with healthy sportsmen. PMID- 11039637 TI - Knee strength and lower- and higher-intensity functional performance in older adults. AB - PURPOSE: This study characterizes the linear relations among knee strength, work capacity, and lower- and higher-intensity measures of functional performance in ambulatory, high-functioning older adults. METHODS: Sixty-two seniors (average age = 73.4 +/- 7.3 yr) participated in the study. Isokinetic measures included the peak flexion/extension torque produced during five continuous repetitions and the total flexion/extension work performed during 20 repetitions (60 degrees x s( 1)). Functional measures included lower-intensity tests (timed 8-foot and 50-foot walking tests at the participants' "normal" pace, and a standing reach task) and higher-intensity tests (a timed 50-foot "brisk" walk, timed chair stands, and a timed stair climb). RESULTS: Isokinetic strength and work capacity measures explained between 41% and 54% of the variance in the higher-intensity functional models and only between 31% and 33% of the variance in the lower-intensity models. The strength of the associations, approximated by the beta coefficients of the strength and work terms, was also greater for the higher-intensity functional tasks. CONCLUSIONS: Further research is warranted to determine whether exercises that increase knee strength and work capacity, improve brisk walking, stair climbing, and chair standing capabilities in older adults. PMID- 11039638 TI - Patterns of glenohumeral joint laxity and stiffness in healthy men and women. AB - PURPOSE: The purpose of this study was to identify gender-related differences in glenohumeral (GH) joint laxity, stiffness, and generalized joint hypermobility in healthy men and women. METHODS: Fifty-one healthy men and women were tested for generalized joint hypermobility, and anterior-posterior (AP) joint laxity and stiffness using a single-group factorial design. RESULTS: Women exhibited significantly more anterior joint laxity (men 8.3 +/- 2.2 mm vs women 11.4 +/- 2.8 mm, P < 0.001) and less anterior joint stiffness (men 20.5 +/- 5.0 N x mm(-1) vs women 16.3 +/- 4.2 N x mm(-1), P < 0.01) than men. Men had significantly more posterior joint laxity than anterior (Ant 8.3 +/- 2.2 mm vs Post 9.6 +/- 2.9 mm; P < 0.001), and women also had significantly less anterior joint stiffness than posterior [Ant 16.3 +/- 4.2 N x mm(-1) vs Post 22.1 +/- 6.9 N x mm(-1); P < 0.01], Women also demonstrated significantly more generalized joint hypermobility than men (men 1.0 +/- 1.7 vs women 2.9 +/- 2.1; P = 0.02). CONCLUSIONS: Our findings may indicate a possible increased risk for instability in women, especially those participating in sports that require repetitive overhead throwing motion. Future investigations should seek to determine the contribution of increased GH joint laxity and decreased joint stiffness to various injury states and examine these variables in other populations such as overhead-throwing athletes. PMID- 11039639 TI - Utilizing exercise to affect the symptomology of fibromyalgia: a pilot study. AB - Fibromyalgia (FM), a rheumatological disorder of unknown origin, is characterized by both physical and psychological symptoms. Although inconclusive results have been reported for most treatment modalities, exercise appears to have universal support for decreasing the myriad of symptoms associated with FM. Weaknesses in the literature, however, prevent conclusive statements regarding exercise prescription and concomitant impact on FM symptomology. PURPOSE: The current pilot study attempted to examine the effect of a 24-wk walking program at predetermined intensities on FM. METHODS: Initial design was a randomized control trial with high- and low-intensity exercise groups, and a control group. Subsequent nonrandomized control trials were based on actual exercise behavior. RESULTS: No differences between initial groups were identified. By collapsing groups, heart rate (HR) decreased (P < 0.05) weeks 0-12. Functional impairments were reduced 54% weeks 0-24, with exercise having a large impact (omega2 = 0.30) on this decrease. By reassigning groups, impact of FM on current health status decreased in the low-intensity group (P < 0.05) and increased in the high intensity group (P < 0.02) weeks 0-24. Omega squared indicated strong influence of exercise on pain (omega2 = 0.51), with greater pain in the high-intensity group. CONCLUSIONS: A larger number of subjects and direct supervision of the training program to increase compliance is necessary to clarify the effects of a walking program on the manifestations of FM. Results indicate that intensity of the walking program is an important consideration. Individuals with FM can adhere to low-intensity walking programs two to three times per week, possibly reducing FM impact on daily activities. PMID- 11039640 TI - Is prepubertal growth adversely affected by sport? AB - PURPOSE: To study the effect of genetic factors, birth weight, early childhood growth, sport, hours of training, and pubertal status on the stature and body mass index (BMI) of children aged 9-13 participating in sports at a competitive level. METHODS: A total of 184 children (96 girls, 88 boys), competing in swimming, tennis, team handball, and gymnastics, were investigated, assessing their height, weight, pubertal development, and BMI. Of these, 137 (76 girls, 61 boys) returned a questionnaire, which enabled us to determine height and BMI at age 2-4, birth weight, and parental heights. RESULTS: Significant differences in standard deviation scores (SDS) for actual height and for height at age 2-4 were found in both sexes between the four sports. In girls, BMI SDS was significantly different between the four sports, whereas no difference was found in boys. Each sport investigated separately showed no change in height SDS and BMI SDS between ages 2-4 and 9-13. A regression analysis showed that target height, height at age 2-4, and pubertal status had a significant impact on actual height. Interestingly, the type of sport and hours of training per week had no effect on height SDS. In boys, BMI at age 2-4 and pubertal status had a significant effect on actual BMI, whereas in girls, only BMI at age 2-4 was significant. CONCLUSIONS: The results suggest that prepubertal growth is not adversely affected by sport at a competitive level and that constitutional factors are of importance for choice of sport in children. PMID- 11039641 TI - Exercise and tumor development in a mouse predisposed to multiple intestinal adenomas. AB - Epidemiological evidence suggests that physical activity may be protective against the development of colon cancer. Potential mechanisms remain largely unexplored due to the paucity of appropriate experimental models. PURPOSE: The purpose of this study was to examine the effect of exercise training on polyp development in an induced mutant mouse strain predisposed to multiple intestinal neoplasia (Min mouse). METHODS: Three-week-old male and female heterozygotes were randomly assigned to control (CON; 10 males, 6 females) or exercise (EX; 11 males, 11 females) groups. In the first week, EX mice were acclimated to treadmill running at 10-18 m x min(-1) for 15-60 min x d(-1). From 4-10 wk of age, mice ran at 18-21 m x min(-1) for 60 min. CON mice sat in Plexiglas lanes suspended above the treadmill for the same time periods. At 10 wk of age, the mice were sacrificed and the intestines removed, opened, and counted for polyps. RESULTS: Skeletal muscle oxidative capacity increased with training as shown by a 64% increase in citrate synthase activity in the gastrocnemius/soleus muscle of EX compared with CON (P = 0.009). There were no significant effects of exercise in the males and females combined on small intestine, colon, or total intestinal polyps (P > 0.05). When analyzed separately, however, there were fewer colon and total polyps in the EX than in the CON males, although the difference was not statistically significant (P = 0.06). CONCLUSIONS: These results suggest that seven weeks of exercise training do not affect the development of intestinal polyps in the Min mouse. Further studies are required to determine if a true sex difference exists or if variations on the current training protocol may affect tumor outcomes. PMID- 11039642 TI - A polymorphism in the alpha2a-adrenoceptor gene and endurance athlete status. AB - PURPOSE: In a case control study, we examined the allelic frequencies and genotype distributions of two restricted fragment length polymorphisms (RFLP) in the alpha-2A-adrenoceptor gene (ADRA2A) and beta-2-adrenoceptor gene (ADRB2) among elite endurance athletes (EEA) and sedentary controls (SC). METHODS: The EEA group included 148 Caucasian male subjects recruited on the basis that they had a VO2max > 74 mL O2 x kg(-1) x min(-1). The SC group comprised 149 unrelated sedentary male subjects, all Caucasians, from the Quebec Family Study. After digestion with the restriction enzymes Dra I (ADRA2A) and Ban I (ADRB2), Southern blotting and hybridization techniques were used to detect the mutations in the two ADR genes, which are encoded on chromosomes 10 (q24-26) and 5 (q31-32), respectively. RESULTS: For the Dra I ADRA2A RFLP, we observed a significant difference in genotype distributions between the two groups (P = 0.037). A higher frequency of the 6.7-kb allele was observed in the EEA group compared with the SC group (P = 0.013). No statistically significant difference was found between groups for the Ban I ADRB2 polymorphic site. Genotype frequencies for both genes in both groups were in Hardy-Weinberg equilibrium. CONCLUSIONS: In summary, we found evidence that ADRA2A gene variability detected with Dra I is weakly associated with elite endurance athlete status, and we conclude that genetic variation in the ADRA2A gene or a locus in close proximity may play a role in being able to sustain the endurance training regimen necessary to attain a high level of maximal aerobic power. PMID- 11039643 TI - Change in left atrial and ventricular dimensions during and immediately after exercise. AB - PURPOSE: The aim of this study was to evaluate differences in the left atrial (LAD), total ventricular end-diastolic (TEDD), end-systolic diameters (TESD), and left ventricular shortening fraction (SF) compared with heart rate (HR) and systolic blood pressure (SBP) during exercise and recovery. METHODS: Healthy young male (N = 15) and female (N = 16) subjects performed an incremental cycle ergometer test in upright position, and three phases of energy supply were defined by means of blood lactate concentration (LA) and respiratory gas exchange variables (I: aerobic; II: aerobic-anaerobic transition; III: anaerobic). Subjects were required to rest their arms on a steering bar and to lean their upper body forward; two dimensional (2-D) echocardiograms were obtained over the left parasternal area at rest (R), at the end of each phase, immediately within 15 s post, and 6 min after exercise (6 min). By using VINGMED's "Anatomical M Mode," it was possible to extract M-Mode Sweeps from stored 2-D-Loops and perform the M-Mode measurement. RESULTS: In contrast to the significant decrease in TEDD and TESD from III to 15 s up to resting values and the significant increase in SF from III to 15 s, the moderate decrease in HR immediately post exercise (15 s) was not significant. The SBP showed a significantly decrease from III to 15 s; in contrast to TEDD, TESD, and SF, the values at 15 s were comparable with the values at II. For LAD, significant increase during exercise and a decrease during recovery were observed. Sex-specific differences of changes in measured variables could not be found. CONCLUSION: We concluded that post exercise measurement of left ventricular and atrial dimensions or SF were not valid to describe heart function at maximal exercise although immediately post exercise HR was near maximal level. PMID- 11039644 TI - Effect of static and dynamic exercise on heart rate and blood pressure variabilities. AB - PURPOSE: This study examines the effect of static and dynamic leg exercises on heart rate variability (HRV) and blood pressure variability (BPV) in humans. METHODS: 10 healthy male subjects were studied at rest, during static exercise performed at 30% of maximal voluntary contraction (SX30), and during dynamic cycling exercises done at 30% of VO2max (DX30) and at 60% of VO2max (DX60). Respiration, heart rate, and blood pressure signals were digitized to analyze temporal and spectral parameters involving short and overall indexes (SD, deltaRANGE, RMSSD, Total power), power of the low (LF), middle (MF), and high (HF) frequency components, and the baroreceptor sensitivity by the alphaMF index. RESULTS: During SX30, indexes of HRV as SD, deltaRANGE, Total power, and MF in absolute units increased in relation with rest values and were significantly higher (P < 0.001) than during DX30 and DX60; HF during SX30, in normalized and absolute units, was not different of the rest condition but was higher (P < 0.001) than HF during DX30 and DX60. Parameters of BPV as SD and deltaRANGE increased (P < 0.001) during both type of exercises, and significant (P < 0.01) increments were observed on MF during SX30 and DX30; systolic HF was attenuated during DX30 (P < 0.05), whereas diastolic HF was augmented during DX60 (P < 0.001). Compared with rest condition, the alphaMF index decreased (P < 0.01) only during dynamic exercises. CONCLUSION: Because HRV and BPV response is different when induced by static or dynamic exercise, differences in the autonomic activity can be advised. Instead of the vagal withdrawal and sympathetic augmentation observed during dynamic exercise, the increase in the overall HRV and the MF component during static exercise suggest an increased activity of both autonomic branches. PMID- 11039645 TI - Relation between heart rate variability and training load in middle-distance runners. AB - PURPOSE: Monitoring physical performance is of major importance in competitive sports. Indices commonly used, like resting heart rate, VO2max, and hormones, cannot be easily used because of difficulties in routine use, of variations too small to be reliable, or of technical challenges in acquiring the data. METHODS: We chose to assess autonomic nervous system activity using heart rate variability in seven middle-distance runners, aged 24.6 +/- 4.8 yr, during their usual training cycle composed of 3 wk of heavy training periods, followed by a relative resting week. The electrocardiogram was recorded overnight twice a week and temporal and frequency indices of heart rate variability, using Fourier and Wavelet transforms, were calculated. Daily training loads and fatigue sensations were estimated with a questionnaire. Similar recordings were performed in a sedentary control group. RESULTS: The results demonstrated a significant and progressive decrease in parasympathetic indices of up to -41% (P < 0.05) during the 3 wk of heavy training, followed by a significant increase during the relative resting week of up to +46% (P < 0.05). The indices of sympathetic activity followed the opposite trend, first up to +31% and then -24% (P < 0.05), respectively. The percentage increasing mean nocturnal heart rate variation remained below 12% (P < 0.05). There was no significant variation in the control group. CONCLUSION: This study confirmed that heavy training shifted the cardiac autonomic balance toward a predominance of the sympathetic over the parasympathetic drive. When recorded during the night, heart rate variability appeared to be a better tool than resting heart rate to evaluate cumulated physical fatigue, as it magnified the induced changes in autonomic nervous system activity. These results could be of interest for optimizing individual training profiles. PMID- 11039646 TI - Oxygen uptake in one-legged and two-legged exercise. AB - PURPOSE: The purpose of this study was to determine the primary factors causing the differential oxygen uptake (VO2) response at submaximal intensities between one-legged and two-legged exercise, and whether peak oxygen uptake (VO2peak) increases in proportion to the increase in active muscle mass. METHODS: Two different types of exercise were used for this experiment, each requiring a different movement, a different method of stabilizing posture, and, finally, a different limiting VO2peak. In experiment 1, nine male subjects performed one legged cycling (OLC) and two-legged cycling exercise (TLC) at a pedaling rate of 80 rpm. The exercise intensity was first set at 80 W and was increased by 40 W every 3 min until exhaustion. In experiment 2, six healthy male subjects performed one-legged knee-extension (OKE) and two-legged knee-extension (TKE) exercise at a rate of 50 contractions per minute. The knee-extension exercise was done at constant work rates for a 3-min session in OKE or a 4-min session in TKE. The exercise bouts were performed intermittently at four to seven different submaximal intensities and VO2 was determined at each intensity in all exercises. RESULTS: At submaximal intensities, VO2 in relation to work rate of one-legged exercise was more steep than those of two-legged exercise, and the mean values of VO2 were significantly higher in one-legged exercise than those in two-legged exercise in both knee extension and cycling exercise. Mean values of VO2peak for two-legged exercise were significantly higher than that for one-legged exercise (P < 0.01); however, it was much lower than two times of that for one-legged exercise even in knee extension exercise where the VO2peak would be limited peripherally. CONCLUSION: The findings of this study suggest that the differential VO2 response between one-legged and two-legged exercise would be attributed not only to the difference in force application throughout the exercise movement and to the effect of a postural component but also to the inhibited circulatory response caused by the multiple limb exercise. In addition, it was supposed that VO2peak does not increase in proportion to the exercising muscle mass even during smaller muscle activity where the cardiac pumping capacity has not reached its upper limit. PMID- 11039647 TI - Effect of short-duration spaceflight on thigh and leg muscle volume. AB - PURPOSE: Human skeletal muscle probably atrophies as a result of spaceflight, but few studies have examined this issue. Thus, little is known about the influence of microgravity upon human skeletal muscle, nor is it possible to assess the validity of ground based models of spaceflight. This study tested the hypothesis that the magnitude of spaceflight induced muscle atrophy would be a function of flight duration and greater than that of bed rest. METHODS: Three astronauts flew 9, 15, and 16 d in space. Volume of the knee extensor (quadriceps femoris), knee flexor (hamstrings, sartorius, and gracilis), and plantar flexor (triceps surae) muscle groups was measured using magnetic resonance imaging before and after spaceflight and during recovery. The volume of each muscle group in each image was determined by multiplying cross-sectional area by slice thickness. These values were subsequently summed to calculate muscle volume. RESULTS: Volume changes in the knee extensor, knee flexor, and plantar flexor muscle groups ranged from -15.4 to -5.5, -14.1 to -5.6, and -8.8 to -15.9%, respectively. Muscle volume decreases normalized by flight duration ranged from 0.62 to 1.04% x d(-1). These relative changes appeared to be greater than those that we have reported previously for bed rest (Akima et al., J. Gravitat. Physiol. 4:15-22, 1997). CONCLUSIONS: These results suggest that atrophy as a result of at least 2 wk of spaceflight varied among individuals and muscle groups and that the degree of atrophy appeared to be greater than that induced by 20 d of bed rest. PMID- 11039648 TI - Adaptations to a 7-day head-down bed rest with thigh cuffs. AB - PURPOSE: Thigh cuffs were two elastic strips fixed at the upper part of each thigh, which limits the shift of fluid from the legs into the cardio-thoracic region. The purpose of this study was to examine the effects of thigh cuffs on hormonal and plasma volume responses and orthostatic tolerance during a 7-day head-down bed rest (HDBR). METHODS: Orthostatic tolerance, plasma volume, total body water, blood volume-regulating hormones, and hydro-electrolyte responses were measured in eight healthy men (age range, 25-40 yr), using thigh cuffs 10 h daily during 7 d of -6 degrees HDBR. RESULTS: Thigh cuffs worn during HDBR attenuated the decrease in plasma volume observed after HDBR (thigh cuffs: -5.85 +/- 0.95% vs control: -9.09 +/- 0.82%, P < or = 0.05). During this experiment, there was no significant change in total body water. Thus, the hypovolemia did not result from a loss of water but from a fluid shift from the blood compartment into the interstitial and/or intracellular compartment. Hormonal responses during HDBR and stand test were not modified by the thigh cuffs. Thigh cuffs had no significant effect on the clinical symptoms of orthostatic intolerance after HDBR. CONCLUSIONS: Thigh cuffs worn during HDBR blunted the decrease in plasma volume but did not reduce orthostatic intolerance; thus, they are not a completely effective countermeasure. Furthermore, hypovolemia seems to be necessary but not sufficient to induce orthostatic intolerance after HDBR. PMID- 11039649 TI - Force and EMG power spectrum during eccentric and concentric actions. AB - INTRODUCTION: This study was designed to examine the force and activation levels of elbow flexor muscles during preactivated eccentric, concentric and isometric actions. METHODS: Force, average EMG (aEMG), and the EMG power spectrum were investigated at different constant movement velocities (1 rad x s(-1), 2 rad x s( 1), 3 rad x s(-1), and 4 rad x s(-1)) at different joint angles. RESULTS: Average force at a 110 degree elbow angle was lower and aEMG was higher in concentric actions as compared with eccentric and isometric actions. At a 55 degree elbow angle, there was no difference in aEMG, or it was slightly higher in eccentric actions. MF was higher in the concentric as compared with eccentric actions at the three fastest velocities at the 110 degree elbow angle, whereas no difference was observed at the 55 degree elbow angle. In concentric action, MF was higher in 4 rad x s(-1) in comparison with 1 rads x s(-1) at 110 degree elbow angle. DISCUSSION: These results suggest that it is difficult to maintain the maximal eccentric force throughout the whole range of motion. Maximal EMG activity and frequencies of the EMG power spectrum can be at the same level or lower in eccentric actions as compared with concentric actions, depending on the joint angle and preactivation mode. The results of the EMG power spectrum do not support the concept that in maximal eccentric actions fast units are selectively activated. PMID- 11039650 TI - The role of maximal strength and load on initial power production. AB - PURPOSE: The influence of maximal strength, as measured by the maximal load lifted for one repetition (1RM), on power production in the initial 200 ms of the concentric phase for both rebound and nonrebound movements was investigated. We also investigated the effect of external load upon this relationship. METHODS: Twenty-seven male subjects (21.9 +/- 3.1 yr, 89.0 +/- 12.5 kg) were separated by previously determined bench press IRM into high (100.88 +/- 7.24 kg) and low (72 +/- 6.61 kg) RM groups. Concentric only bench presses and rebound bench presses were compared between and within groups to note the effect of RM across external loads of 40%, 60%, and 80% 1RM, on instantaneous, mean, and peak power output. RESULTS: The results of this study clearly indicated the enhancement of concentric motion by prior eccentric muscle action (336-1332% enhancement in the first 20 ms). Possessing a high RM augmented power production in the initial 200 ms of stretch-shorten cycle activity, across all the external resistances tested (P < 0.05). The temporal characteristics of this enhancement, however, differed across loads. That is, 80% IRM loading showed a later time to peak enhancement (80 ms vs 20 ms). Interestingly, the influence of RM on concentric only motion in the initial 200 ms across the external resistances tested was found to be nonsignificant. CONCLUSIONS: The results suggest that the role of maximal strength during initial power production between concentric and stretch-shorten cycle activity differs, which has important implications for the training of athletes. PMID- 11039651 TI - Effects of back extensor strength training versus balance training on postural control. AB - PURPOSE: Aim of this study was to investigate effects of 1) regular back extensor strength training as opposed to balance training, and 2) the influence of the sequence of both training types on postural control, force, and muscle efficiency. METHODS: Twenty-six young, healthy subjects were investigated at baseline, 1 month and 2 months later. At each examination, subjects completed a posturographic, balance skill, and isometric maximum voluntary (MVC) back extension testing, including surface electromyographic (SEMG) recordings. After baseline evaluation, subjects were assigned to either daily strength training or balance training. After 1 month, the type of training was exchanged between groups. RESULTS: After 1 month, back extensor strengthening led to decreased postural stability on hard surface, whereas there were no change after balance skill training. Analysis of the low- and high-frequency components of the sway signal revealed that strength training increased control efforts as indicated by an increased high-frequency component in order to maintain postural stability and unchanged low-frequency component. Balance skill training, however, increased postural stability as indicated by a decreased low-frequency component. The control effort remained unchanged. After completing either sequence of training, all postural parameters remained unchanged in both groups. Muscular efficiency as measured by SEMG root mean square during a standardized motor skill task revealed improved muscle economy regardless of the type of training. Back extension torque improved in both groups. CONCLUSION: To avoid reduction of postural stability in rehabilitation processes, we recommend to include antagonist muscles in a comprehensive strength training regime or balance skill training. PMID- 11039652 TI - Heart rate and performance parameters in elite cyclists: a longitudinal study. AB - PURPOSE: This study was designed to evaluate the stability of target heart rate (HR) values corresponding to performance markers such as lactate threshold (LT) and the first and second ventilatory thresholds (VT1, VT2) in a group of 13 professional road cyclists (VO2max, approximately 75.0 mL x kg(-1) x min(-1)) during the course of a complete sports season. METHODS: Each subject performed a progressive exercise test on a bicycle ergometer (ramp protocol with workload increases of 25 W x min(-1)) three times during the season corresponding to the "active" rest (fall: November), precompetition (winter: January), and competition periods (spring: May) to determine HR values at LT, VT1 and VT2. RESULTS: Despite a significant improvement in performance throughout the training season (i.e., increases in the power output eliciting LT, VT1, or VT2), target HR values were overall stable (HR at LT: 154 +/- 3, 152 +/- 3, and 154 +/- 2 beats x min(-1); HR at VT1: 155 +/- 3, 156 +/- 3, and 159 +/- 3 beats x min(-1); and at VT2: 178 +/- 2, 173 +/- 3, and 176 +/- 2 beats x min(-1) during rest, precompetition, and competition periods, respectively). CONCLUSION: A single laboratory testing session at the beginning of the season might be sufficient to adequately prescribe training loads based on HR data in elite endurance athletes such as professional cyclists. This would simplify the testing schedule generally used for this type of athlete. PMID- 11039653 TI - Effect of fluid intake volume on 2-h running performances in a 25 degrees C environment. AB - PURPOSE: In this study, we examined the effects of greater than ad libitum rates of fluid intake on 2-h running performances. METHODS: Eight male distance runners performed three runs on a treadmill at 65% of peak oxygen uptake (VO2peak) for 90 min and then ran "as far as possible" in 30 min in an air temperature of 25 degrees C, a relative humidity of 55% and a wind speed of 13-15 km x h(-1). During the runs, the subjects drank a 6.9% carbohydrate (CHO)-electrolyte solution either ad libitum or in set volumes of 150 or 350 mL x 70 kg(-1) body mass (approximately 130 or 300 mL) every 15-20 min. RESULTS: Higher (approximately 0.9 vs 0.4 L x h(-1)) rates of fluid intake in the 350 mL x 70 kg( 1) trial than in the other trials had minimal effects on the subjects' urine production (approximately 0.1 L x h(-1)), sweat rates (approximately 1.2 L x h( 1)), declines in plasma volume (approximately 8%), and rises in serum osmolality (approximately 5 mosmol x L(-1)) and Na+ concentrations (approximately 7 mEq x L( 1)). A greater (approximately 1.0 vs 0.5 g x min(-1)) rate of CHO ingestion in the 350 mL x 70 kg(-1) trial than in the other trials also did not affect plasma concentrations of glucose (> or = 5 mmol x L(-1)) and lactate (approximately 3 mmol x L(-1)) during the performance runs. In all three performance runs, increases in running speeds from approximately 14 to 15-16 km x h(-1) and rises in exercise intensities from approximately 65% to 75% of VO2peak elevated plasma lactate concentrations from approximately 1.5 to 3 mmol x L(-1) and accelerated CHO oxidation from approximately 13 to 15 mmol x min(-1). The only effect of the additional intake of approximately 1.0 L of fluid in the 350 mL x 70 kg(-1) trial was to produce such severe gastrointestinal discomfort that two of the eight subjects failed to complete their performance runs. CONCLUSION: Greater rates of fluid ingestion had no measurable effects on plasma volume and osmolality and did not improve 2-h running performances in a 25 degrees C environment. PMID- 11039654 TI - Reliability of an air-braked ergometer to record peak power during a maximal cycling test. AB - PURPOSE: To assess the reliability of the Kingcycle ergometer, this study compared peak power recorded using a Kingcycle and SRMTM power meters during Kingcycle maximal aerobic power tests. METHODS: The study was completed in two parts: for part 1, nine subjects completed three maximal tests with a stabilizing kit attached to the Kingcycle rig and calibration of the Kingcycle checked against SRM (MAP(C)); and for part 2, nine subjects completed two maximal tests without the stabilizing kit and the Kingcycle calibrated using the standard procedure (MAP(S)). Each MAP(C) test was separated by 1 wk; however, MAPs tests were separated by 54 +/- 32 d, (mean +/- SD). Testing procedure was repeated for each MAP and peak power output was calculated as the highest average power output recorded during any 60-s period of the MAP test using the Kingcycle (King(PPO)) and SRM (SRM(PPO)). RESULTS: Coefficient of variations (CVs) for King(PPO) were larger than those of SRM(PPO); 2.0% (95%CI = 1.5-3.0) versus 1.3% (95%CI = 1.0 2.0) and 4.6% (95%CI = 2.7-7.6) versus 3.6% (95%CI = 2.1-6.0) for MAPC( and MAP(S), respectively. During all tests, King(PPO) was higher than SRM(PPO) by an average of approximately 10% (P < 0.001). CONCLUSIONS: Investigators should be aware of the discrepancy between the two systems when assessing peak power and that SRM cranks provide a more reproducible measure of peak power than the Kingcycle ergometer. PMID- 11039655 TI - Comparison of Actiwatch activity monitor and Children's Activity Rating Scale in children. AB - PURPOSE: The Children's Activity Rating Scale (CARS) is a rating scale that is used in direct observation of physical activity in children. Direct observation is costly and tedious, and accuracy may decrease as the observation period lengthens. Recently, motion sensors have gained acceptance for assessment of physical activity. The purpose of this study was to compare 6 h of activity levels using simultaneous monitoring of preschool aged children with CARS and the Actiwatch (Mini-mitter Company Inc.) activity monitor. METHODS: A total of 40 children had 5-6 h (mean of 5.9 h) of direct observation while wearing a monitor on the waist. Simultaneous 3-min mean CARS scores and 3-min activity counts were matched for each subject. RESULTS: The range for the mean 3-min CARS scores ranged from 1.00 to 4.50. The 3-min activity counts ranged from 0 to 9,695 with a mean of 670 (median 243). The within child correlations between the 3-min CARS score and the 3-min sensor readings ranged from 0.03 to 0.92 (median of 0.74). We found the correlation coefficients were higher in those children who were more active, probably due to the larger ranges in the CARS scores. When using mixed model repeated measures, sensor readings were significantly associated with CARS (P < 0.001). CONCLUSION: Our results indicate that the 3-min CARS score correlates with 3-min activity counts, favoring the use of the activity monitors in assessing physical activity in preschool-aged children. PMID- 11039656 TI - A simple method for individual anaerobic threshold as predictor of max lactate steady state. AB - BACKGROUND: The individual anaerobic threshold (IAT) is defined (18) as the highest metabolic rate where blood lactate (La) concentrations are maintained at a steady-state during prolonged exercise. Stegmann et al.'s (18) method to detect IAT, using La-performance relationship during incremental graded exercise, is based on the assumption that La is in relatively steady state by the end of each 3-min stage of work rate. However, at the end of a 3-min stage, an La steady state (Lass) is not reached (13). PURPOSE: The present study was designed to investigate whether the IAT should be determined by attributing La value to the antecedent stage (IATa) or to the same stage of its measurement (IATm), then to verify whether this IAT would be a valid indicator of the max Lass during prolonged exercise. METHODS: Forty-one athletes (21 male and 20 female), regularly involved in different physical training, performed three exercise tests on treadmill. The first one was a 3-min stage incremental test to detect the IATa and IATm. The other two tests were 30-min prolonged tests at the IATa and IATm workload. Lass were present in IATa intensity (about 4.0 mmol x L(-1)) both in male and female athletes, whereas at IATm intensity a Lass was not present and a premature break-off occurred in some cases. DISCUSSION: This protocol can be useful for practical use because: 1) the method of choosing the anaerobic threshold is easy to apply; 2) it does not require to reach the maximal effort; and 3) although in some cases the IATa could probably underestimate the workload of max Lass, the IATa can be regarded as guideline to define the intensity of endurance training. PMID- 11039657 TI - FT-IR spectroscopy utilization to sportsmen fatigability evaluation and control. AB - PURPOSE: A longitudinal biological study of 20 elite rowers was performed using capillary blood (serum) FT-IR spectra to evaluate their training load adaptations and fatigue. METHODS: Difference spectra (rest serum spectra subtracted to exercise serum spectra) were used to evaluate subjects' metabolic response to exercise. Spectra classifications were used for serum contents differentiation on the basis of biomolecular absorption. RESULTS: For two subjects, several metabolic differentiations were observed. These started with sugars metabolism on the fifth training week, followed successively by lipid metabolism and protein metabolism, when overtraining was clinically diagnosed. Several weeks further into the training program, the same onset of metabolic differentiations was observed for eight other subjects. When differentiations reached lipid metabolism, they were asked to reduce their training loads. Unlike the overtrained subjects, a rapid recovery was observed (3 vs 22 wk) and metabolism alterations disappeared. CONCLUSION: The fatigability limit in sportsmen seemed to be situated at a certain level of metabolic stress, beyond which a rapid overtraining process recover was no longer possible. PMID- 11039658 TI - The 12-lead ECG is the most cost-effective preprecipitation cardiovascular screen. PMID- 11039659 TI - World Health Organization classification of the acute leukemias and myelodysplastic syndrome. AB - Under the auspices of the European Association of Hematopathologists and the Society for Hematopathology, 100 hematopathologists and clinicians have met together and in subcommittees over a 5-year period. The model used was that developed by the International Lymphoma Study Group, which used morphologic, immunologic, and genetic features. It resulted in the revised European-American Lymphoma classification. The newly proposed leukemia classification uses a similar format and eliminates purely morphologic subtypes that have no current clinical relevance. As with all consensus proposals, these classifications must withstand criticism and debate by potential users. PMID- 11039660 TI - Alternatives to conventional or myeloablative chemotherapy in myelodysplastic syndrome. AB - At present, the only 2 treatments that can prolong survival in patients with myelodysplastic syndrome (MDS) are allogeneic stem cell transplantation and intensive chemotherapy. Alternatives to myeloablative or conventional chemotherapy include: (1) supportive therapy, (2) stimulation of normal residual hematopoietic progenitors, and (3) manipulation of myelodysplastic hematopoiesis. These alternative therapeutic strategies can be accomplished using various therapeutic tools. Supportive therapy remains the mainstay in the management of MDS patients and desferrioxamine should be administered to individuals who have a regular need for blood transfusion. The only hematopoietic growth factors that can be useful in the treatment of selected MDS patients are recombinant human erythropoietin (rhEpo) and granulocyte colony-stimulating factor (G-CSF). Overall, 15% to 20% of patients with MDS respond to rhEpo treatment. Factors predicting response include serum erythropoietin levels <100 to 200 mU/mL, low risk MDS, and no or low need for transfusion. G-CSF alone should be used only for short-term treatments during severe infection episodes that do not respond to conventional therapy. About 40% of MDS patients respond to a combined treatment of rhEpo plus G-CSF with amelioration of anemia. Cytoprotective antiapoptotic agents such as amifostine, alone or in combination, may improve blood values in occasional MDS patients. MDS patients with immunologically mediated myelosuppression may respond favorably to antithymocyte globulin or cyclosporin A (CyA). Although rhEpo and CyA may be used in individual patients who appear likely to respond, the remaining therapeutic tools must be considered strictly experimental; phase III clinical trials are required to establish whether they can be useful in the treatment of MDS patients. More generally, because of the current uncertainties concerning MDS treatment, participation of patients in clinical trials should be always encouraged. PMID- 11039661 TI - Intensive chemotherapy for patients with high-risk myelodysplastic syndrome. AB - Standard antileukemic chemotherapy induces complete remission in approximately half of patients with high-risk (HR) myelodysplastic syndrome (MDS). Intensification of induction therapy by the use of intermediate- or high-dose cytosine arabinoside in combination with fludarabine, idarubicin, or topotecan seemingly improved complete response (CR) rates, particularly in patients with poor prognosis karyotypes. The various high-intensity regimens appear to be comparable in inducing CR, although some are better tolerated with low mortality even in advanced-age populations with MDS. The encouraging early results with new agents, eg, topoisomerase inhibitors (topotecan) and hypomethylating agents (5 azacytidine), have been disappointing because long-term follow-up has shown continuous relapses. Regardless of the intensity of chemotherapy, remissions are short, even with continuation of intensive postremission therapy. Long-term disease-free survival remains dismal. In a large population with HR MDS treated with high-dose chemotherapy, only 5% of patients were alive at 3 years, and a majority of survivors were the younger patients with diploid karyotype refractory anemia with excess blasts in transformation. Further intensification of either induction chemotherapy or postremission therapy is unlikely to improve results with current drug combinations. With these results at hand, the role of intensive chemotherapy in the management of MDS remains controversial. Because CR status is associated with clinical benefits and possibly better survival, induction of CR should remain an important aim for HR MDS. The intensive combination chemotherapy may be integrated as an initial part of the management of HR MDS, as an alternative for patients not eligible for allogeneic bone marrow transplantation (BMT). Regimens with low early mortality, eg, topotecan plus intermediate dose Ara-C, could also be used to reduce tumor load prior to allogeneic BMT. Induction of CR should be attempted with the most effective and best tolerated regimens, particularly, but not only, in younger patients with good performance status regardless of karyotype. Postremission therapy remains a major challenge. It should involve either allogeneic BMT or investigational approaches to eliminate or control the minimal residual disease with different mechanisms In elderly patients, allogeneic "minitransplant" is an investigational alternative seeking to exploit graft-versus-tumor reaction. New agents, such as farnesyl transferase inhibitors (ras inhibitors), drug-antibody conjugates (Myelotarg), thalidomide, arsenic trioxide, maintenance chemotherapy with hypomethylating agents, or oral topoisomerase inhibitors along with agents modulating factors affecting growth and differentiation, should be explored to maintain remission with minimal compromise of quality of life. Although prognostic scoring systems such as the new International Prognostic Scoring System (IPSS) do not predict for initial response, IPSS continues to predict survival in patients treated with high-dose chemotherapy. Although activity of new agents could be rapidly assessed in single arm pilot studies, the final merit of high-dose chemotherapy could be assessed only in well-designed randomized trials with patients assigned according to risk based classification and evaluated for response by generally accepted and standardized criteria. PMID- 11039662 TI - Stem cell transplantation for patients with myelodysplastic syndrome and secondary leukemias. AB - Most patients with myelodysplastic syndrome (MDS) are too old to be considered for intensive treatment, such as stem cell transplantation (SCT). Allogeneic SCT from an HLA-identical sibling donor is the curative treatment option for a relatively young patient (younger than 60 years) with MDS or secondary acute myeloid leukemia. Older age and lack of sibling donors limit this application. Alternative stem cell sources, such as unrelated donors, nonidentical family members, or autologous transplants, have been used more recently. Most patients may benefit optimally from an allogeneic SCT when the transplant is performed as soon as an HLA-identical family member has been identified. Progression to more advanced leukemia conditions will be associated with a higher failure rate due to increased relapse rate after SCT and higher treatment-related mortality. Delay of the transplant may be justified in a minority of patients with refractory anemia or refractory anemia with ringed sideroblasts without profound cytopenias or complex cytogenetic abnormalities and no need for erythrocyte transfusions. The present data from patients transplanted with sources of hematopoietic stem cells other than histocompatible sibling donors give an indication of the potential of other forms of transplantation. The disease-free survival of patients transplanted with histocompatible sibling donors was significantly better than the outcome of patients transplanted with other sources of stem cells. About one third of the patients transplanted with stem cells from histocompatible siblings and about one-quarter of the patients with stem cells from other sources may be free of disease for 3 years or longer. The results of these treatment forms have improved considerably, but the continuing high treatment-related mortality warrants that these patients should be treated within investigational protocols. PMID- 11039663 TI - Animal models for X-linked sideroblastic anemia. AB - Erythroid 5-aminolevulinate synthase (ALAS-E) catalyzes the first step of heme biosynthesis in erythroid cells. Several lines of evidence suggest that the expression of ALAS-E is important for the process of erythroid differentiation, which requires a large amount of heme for hemoglobin production. Mutation of human ALAS-E causes the disorder X-linked sideroblastic anemia (XLSA). More than 25 unrelated ALAS-E mutations in XLSA patients have been reported. Most XLSA cases are of the pyridoxine-responsive type, but molecular diagnosis of 1 pyridoxine-refractory type XLSA has also been reported. To examine the roles heme plays during hematopoiesis and to create animal models of XLSA, we disrupted the mouse ALAS-E gene. A chemically induced zebrafish mutant (sau) that lacks ALAS-E has also been isolated. Analysis of these ALAS-E mutants unequivocally demonstrated that ALAS-E is the principal isozyme contributing to erythroid heme biosynthesis In ALAS-E-null mutant mouse embryos, erythroid differentiation was arrested, and an abnormal hematopoietic cell fraction emerged that accumulated a large amount of iron diffusely in the cytoplasm. This accumulation of iron was in contrast to that in XLSA patients, as typical ring sideroblasts accumulated iron primarily in mitochondria. These observations suggest that the mode of iron accumulation caused by the lack of ALAS-E is different in primitive and definitive erythroid cells. Thus ALAS-E, and hence heme supply, is necessary for erythroid cell differentiation and iron metabolism. PMID- 11039664 TI - Polycomb-group genes and hematopoiesis. AB - Although long-range chromatin organization during cell differentiation has not yet been determined, considerable evidence suggests that regulation of the chromatin structure may play a crucial part in transcriptional regulation. Drosophila genetics has introduced a unique system that can maintain genes in the on or off state after the initial gene expression decision has been established. This maintenance system is known to be mediated through the trithorax-group (trxG) and Polycomb-group (PcG) genes. The products from these 2 genes individually form multimeric complexes in the chromatin. The trxG genes are known to maintain the transcriptionally active states of the chromatin, and the PcG genes are thought to maintain the repressive states. The function of the PcG genes is defined in terms of anteroposterior patterning not only in Drosophila but also in mammals, whereas mammalian PcG genes have additional functions in higher order biological functions All PcG gene-deficient mice have provided evidence that these genes play a crucial role in hematopoiesis These findings should help shed new light on the roles of the chromatin regulatory system in hematopoiesis. PMID- 11039665 TI - Thrombopoietin and myelodysplastic syndromes. AB - Thrombopoietin (TPO), a major cytokine involved in megakaryocystopoiesis/thrombopoiesis, may be effective for the treatment of thrombocytopenia associated with myelodysplastic syndromes (MDS). We reviewed the available data relating to the therapeutic potential of TPO for MDS and found the following. The endogenous TPO level is elevated in MDS patients, especially in those with refractory anemia (RA). In RA patients, but not in patients with RA with excess blasts (RAEB) or RAEB in transformation (RAEB-t), both the platelet and megakaryocyte counts correlate inversely with the endogenous TPO level. This scenario indicates that the physiological mechanism for regulating the endogenous TPO level is conserved, at least in part, in RA patients. The number of TPO receptors (TPO-R) expressed on platelets and CD41+ and/or CD34+ cells in MDS is reduced to nearly half the number present in normal subjects. This is consistent with the finding that TPO-induced in vitro megakaryocytopoiesis is not uniformly observed in MDS. Meanwhile, in some patients with RAEB, RAEB-t, or chronic myelomonocytic leukemia, blasts have the TPO-R mRNA and probably TPO-R protein. This fact may explain the lack of correlation between the endogenous TPO level and the platelet and megakaryocyte counts in RAEB and RAEB-t and suggests that TPO may induce blast proliferation in some cases. These findings may be of use when designing a clinical trial of TPO for MDS. PMID- 11039666 TI - Residual erythroid progenitors in W/W mice respond to erythropoietin in the absence of steel factor signals. AB - Erythropoiesis is severely impaired in mice with inactivating mutations in the Steel factor (SF) gene (Sl/Sl mice) or in c-kit, in the SF receptor gene (W/W mice), and in mice with null mutations in the genes for either erythropoietin (EPO) or the erythropoietin receptor (EPO-R). Previous studies indicated that EPO is sufficient for colony development from colony-forming units-erythroid (CFU-E). However, recent studies have shown that there is a physical association between these 2 receptors and that c-kit can phosphorylate EPO-R. To examine the role SF and EPO play in regulating erythropoiesis, we examined the effect of SF and EPO on colony development from cells of the embryonic aorta-gonad-mesonephros (AGM) region, yolk sac, and liver of fetal wild-type and W/W mice. The maturation of CFU-E from these sites did not require the addition of SF to clonal cultures, whereas the efficient development of erythroid bursts required both EPO and SE The number of erythroid colony-forming cells was reduced in both the AGM region and liver of fetal W/W mice. The residual CFU-E present in W/W mice were dependent on EPO and independent of SF. These results indicate that EPO/EPO-R can function to support colony formation in the absence of an SF signal. PMID- 11039667 TI - Serum-soluble c-kit levels during mobilization of peripheral blood stem cells correlate with stem cell yield. AB - c-Kit is expressed in hematopoietic stem cells and plays an important role in hematopoiesis. In 16 patients with malignancies, serum-soluble c-Kit levels and the expressions of c-KIT messenger RNA (mRNA) and protein in peripheral blood mononuclear cells were analyzed serially during 26 courses of peripheral blood stem cell (PBSC) mobilization after granulocyte colony-stimulating factor administration following chemotherapy for PBSC harvest. Serum-soluble c-Kit levels were significantly lower in patients than in controls (179.7+/-17.7 arbitrary units [AU]/mL versus 274.5+/-18.9 AU/mL; P < .001), decreasing after chemotherapy (167.7+/-18.2 AU/mL), increasing from day 14, and peaking at day 19 (193.3+/-16.4 AU/mL). The numbers of both c-Kit+ cells and CD34+ cells and granulocyte-macrophage colony-forming units in peripheral blood peaked at day 17, following the peak of the expression of c-KIT mRNA. Serum-soluble c-Kit levels showed a significant positive correlation with the numbers of CD34+ cells in both peripheral blood and leukapheresis products (r = 0.553, P < .01, and r = 0.640, P < .001, respectively) and changed at higher levels in patients with large numbers of PBSCs versus patients with small numbers of PBSCs (P < .05). Serum-soluble c Kit may reflect the capacity for hematopoiesis after chemotherapy and may be useful in predicting the number of PBSCs that can be mobilized and harvested after mobilization, as well as for monitoring the timing for PBSC harvest. PMID- 11039668 TI - Analysis of serum lactate dehydrogenase and its isozymes during administration of granulocyte colony-stimulating factor in children. AB - Serum lactate dehydrogenase (LDH) activity is known to become elevated following granulocyte colony-stimulating factor (G-CSF) therapy in some patients, but no extensive studies on this phenomenon have been performed. In 26 children with malignancies in complete remission who received recombinant human G-CSF intravenously after combined chemotherapy, we measured serum LDH and its isozymes (81 episodes) before chemotherapy (pre-Tx), during administration of G-CSF (mid Tx), and after stopping G-CSF (post-Tx) and compared the obtained data using paired t test. Twelve episodes were excluded from analysis because of liver dysfunction (alanine aminotransferase > 45 IU/L). Serum LDH at mid-Tx (343.1+/ 19.8 IU/L; mean +/- SE) was significantly higher than that at pre-Tx (186.9+/-6.7 IU/L) and post-Tx, but this difference was observed only when change in white blood cell counts (WBCs) (WBC at mid-Tx minus WBC at pre-Tx) was greater than or equal to 4000/microL (58 episodes). Percentages of LDH isozymes 3, 4, and 5 at mid-Tx (23.5+/-1.0, 11.7+/-0.8, and 8.3+/-0.7) were significantly increased compared with those at pre-Tx (19.5+/-0.7, 6.3+/-0.3, and 4.2+/-0.5) and post-Tx, resulting in a significant decrease in percentages of LDH isozymes 1 and 2 at mid Tx. In episodes of change in WBCs > or =4000/microL, change in LDH significantly correlated to changes in WBCs and granulocytes but not to change in lymphocytes or monocytes. These results suggest that serum LDH is significantly elevated during G-CSF administration in accordance with the increase in peripheral granulocytes, which accompanies change in the pattern of percentages of LDH isozymes. PMID- 11039669 TI - Combination chemotherapy with risk factor-adjusted dose attenuation for high-risk myelodysplastic syndrome and resulting leukemia in the multicenter study of the Japan Adult Leukemia Study Group (JALSG): results of an interim analysis. AB - Forty-nine adult patients with high-risk myelodysplastic syndrome (MDS) or acute myeloid leukemia that progressed from MDS were registered for the multicenter study of the Japan Adult Leukemia Study Group. Forty-three patients were evaluable for the analysis. Idarubicin 12 mg/m2 per day for 3 days and continuous cytosine arabinoside 100 mg/m2 per day for 7 days were given as induction therapy, followed by postremission chemotherapy after complete remission (CR). Because elderly patients and those with hypoplastic marrow usually have complications after intensive chemotherapy, often causing early death, the treatment dose was reduced to 60% or 80% according to the presence of 3 risk factors: age 60 years or older, performance status 2 or more, or presence of hypoplastic bone marrow. Of the 43 evaluable patients (median age, 58 years), 26 (60%) achieved CR. Two patients (5%) died within 2 months of completion of induction therapy. The CR rates for patients treated with 100%, 80%, and 60% of the chemotherapy dose were 55% (12 of 22), 63% (10 of 16), and 80% (4 of 5), respectively, indicating that the risk factor-adjusted dose attenuation was appropriately applied to those who might have had problems with the original dose, thus reducing regimen-related mortality rate. The median overall survival of the 43 patients was 8 months. PMID- 11039670 TI - Two cases showing clonal progression with full evolution from aplastic anemia paroxysmal nocturnal hemoglobinuria syndrome to myelodysplastic syndromes and leukemia. AB - We report 2 paroxysmal nocturnal hemoglobinuria (PNH) patients who were initially diagnosed with aplastic anemia and sequentially developed PNH, myelodysplastic syndromes (MDS), and leukemia. Flow cytometry and cytogenetic analysis showed the initial appearance and expansion of PNH clones, gradual replacement of PNH clones by MDS clones with monosomy 7, and then expansion of MDS clones or their subclones with additional chromosomal abnormalities. In relation to these developments, expression increased of the Wilms' tumor gene WT1, a marker for leukemic progression. These patients not only shared bone marrow failure but also might have harbored a hematopoietic environment favorable for the emergence of abnormal clones leading to leukemogenesis. PMID- 11039671 TI - Frequency of a 30-base pair deletion in the latent membrane protein-1 gene in Epstein-Barr virus-associated lymphomas in Japan. AB - The association of Epstein-Barr virus (EBV) with various lymphoid malignancies has been reported. The precise pathogenesis of EBV in malignancies has not yet been elucidated. Latent membrane protein-1 (LMP-1) and Epstein-Barr nuclear antigen-2 (EBNA-2) genes are suspected to be tumorigenic genes. Previous studies suggest that a deletion within the LMP-1 gene may increase the oncogenic potential of EBV. In this study, we analyzed the sequence within the carboxy terminal end of the LMP-1 gene in paraffin-embedded specimens from T-cell lymphomas, Hodgkin's disease (HD), and the buffy coat of peripheral blood from healthy individuals in Japan. Polymerase chain reaction (PCR) was performed using primers spanning the carboxy terminal region of the LMP-1 gene, and sequence analysis was performed to show the exact location of the deletion. The PCR product of the Raji cell line was 161 base pairs (bp), and the LMP-1 gene with deletion was 30 bp shorter in a direct sequence of PCR products. The 30-bp deletion was located in position 168285-168256 of the Raji cell. A deletion within the LMP-1 gene was found in 4 of 25 cases (16%) of EBV-positive T-cell lymphomas, 4 of 10 cases (40%) of EBV-positive HD cases, and 2 of 13 specimens (15%) with amplified PCR products from 49 healthy individuals. The incidence of the 30-bp deletion within the LMP-1 gene in HD was comparable to that of subjects in the United States and Brazil, but the deletion was not found in a high proportion of EBV-positive T-cell lymphoid malignancies. No statistical significance was found regarding the clinical outcome between patients with a deletion within the LMP-1 gene and patients with wild-type LMP. This deletion cannot be considered as simply causing the pathogenesis of EBV-associated lymphoid malignancies in Japan. PMID- 11039672 TI - Cellular biological differences between human myeloma cell lines KMS-12-PE and KMS-12-BM established from a single patient. AB - To clarify cellular biological varieties of myeloma cells, biological differences were analyzed between 2 human myeloma cell lines, KMS-12-PE and KMS-12-BM, derived from pleural effusion and bone marrow, respectively, of a single patient. Although both lines were considered to be derived from the same clone because both had the same chromosomal marker and immunoglobulin H rearrangement, several biological differences were noted. CD11a and CD20 were highly expressed in the KMS-12-BM line, whereas the KMS-12-PE line showed a higher expression of CD7 and CD95/Fas. Although growth was stimulated in KMS-12-BM by interleukin-6 and interferon-alpha, it was inhibited in KMS-12-PE. In addition, apoptosis inhibitors Bcl-2 and Bcl-X(L) were highly expressed in KMS-12-BM cells. Because KMS-12-PE was cultivated 2 months before KMS-12-BM, these differences might be related to their origin (pleural effusion and bone marrow) or the phases of disease progression. However, these biological differences may help clarify myeloma cell biology and lead to improvement in treatment for myeloma patients. PMID- 11039673 TI - Hypersensitivity to mosquito bites is not an allergic disease, but an Epstein Barr virus-associated lymphoproliferative disease. AB - Recently, we showed that 5 cases of hypersensitivity to mosquito bites (HMB) concealed the clonal lymphoproliferation of Epstein-Barr viral (EBV) DNA-positive natural killer (NK) cells. Although the symptoms of HMB have been supposed to derive from Arthus phenomenon, it has become apparent that this unique disorder has the potential to develop into so-called malignant histiocytosis (MH) or related disorders. Accordingly, the criteria for MH have been changed, and a newer diagnostic name, hemophagocytic syndrome, has been described as being associated with viral infection or leukemia/lymphoma. We previously reported that biopsy specimens taken from skin lesions demonstrated infiltration of lymphocytes bearing the phenotype of NK cells. In this study, we found that skin lesions exhibited infiltration of atypical lymphocytes around the small vessels, resembling angiocentric lymphoma, and that these infiltrating cells were positive for EBER-1 by in situ hybridization. These findings support the concept that HMB is the most important manifestation of a certain type of lymphoproliferative disease that presents with an intense local skin reaction and high fever following mosquito bites, and whose essence is the lymphoproliferation of EBV DNA positive NK cells. PMID- 11039674 TI - Multicenter prospective study of interferon-alpha and conventional chemotherapy versus bone marrow transplantation for newly diagnosed patients with chronic myelogenous leukemia. Kouseisho Leukemia Study Group. AB - We compared interferon-alpha (IFN-alpha therapy with bone marrow transplantation (BMT) after initial conventional chemotherapy in patients with chronic myelogenous leukemia (CML) in a multicenter prospective study. Ninety patients with Philadelphia chromosome-positive CML in chronic phase were enrolled between 1991 and 1994. Sixty-six of 89 evaluable patients received IFN-alpha after conventional chemotherapy with hydroxyurea or busulfan (IFN-alpha group). Twenty three patients received allogeneic BMT (BMT group). Fifteen of them received transplants from HLA-identical family donors and 8 from HLA-matched unrelated donors. Forty-seven of 66 patients (71%) in the IFN-alpha group and 17 of 23 patients (74%) in the BMT group achieved complete hematologic response, and 12% in the IFN-alpha group and 13% in the BMT group achieved partial hematologic response. Complete cytogenetic response was induced in 5 (8%), partial cytogenetic response in 8 (12%), and minor cytogenetic response in 12 (18%) in the IFN-alpha group. At a median follow-up of 54 months (range, 30-76 months), in the IFN-alpha group, the predicted 6-year survival rate was 54.5% and the predicted 6-year rate of those remaining in chronic phase was 45.7%. Compared with patients with no cytogenetic response, the patients with some cytogenetic response after IFN-alpha treatment had significantly superior survival and duration of the chronic phase even after correction for the time to response using landmark analysis (P < .05). In the BMT group, the predicted 5-year survival rate was 93.3% for family-donor BMT and 21.9% for unrelated-donor BMT Acute graft-versus-host disease of grade III or IV was observed in 1 of 15 patients who received family-donor BMT and 3 of 8 patients who received unrelated donor BMT. Prior treatment with conventional cytotoxic drugs induced early hematologic response and did not reduce the effect of IFN-alpha on CML. Unrelated donor transplantation should be offered to some patients according to patient age, HLA-matching status, time from diagnosis to BMT, and risk factors. PMID- 11039675 TI - Effective and safe interferon treatment for Japanese patients with chronic myelogenous leukemia relapse after bone marrow transplantation. The Nagoya Blood and Marrow Transplantation Group. AB - The purpose of this study was to assess the efficacy and safety of interferon (IFN) treatment in patients with a relapse of chronic myelogenous leukemia (CML) after bone marrow transplantation in Japan. Accordingly, we retrospectively analyzed the results obtained from 8 patients treated with IFN by the Nagoya Blood and Marrow Transplantation Group. One of 3 patients with hematologic relapse and all 5 patients with cytogenetic relapse achieved complete cytogenetic response (CCR). The median time to achieve CCR was 8 months (range, 3-16 months). One patient relapsed 9 months after starting IFN and died of blast crisis. CCR was maintained for a median duration of 47 months (range, 9-79 months) in the remaining 5 patients. The median duration of survival of these 5 patients after starting IFN was 58 months (range, 12-89 months). At the time of this report, 2 patients who did not attain CCR have survived for 81 months and 142 months after starting IFN, respectively. During IFN treatment, 1 patient showed a transient deterioration of chronic graft-versus-host disease, and no treatment-related deaths were observed. These results suggest that treatment with IFN for CML patients who relapse after bone marrow transplantation is effective and safe. A prospective study to compare IFN with donor lymphocyte infusion is necessary to establish the optimal strategy for the treatment of CML patients who relapse after bone marrow transplantation. PMID- 11039676 TI - Early onset of hemophagocytic syndrome following allogeneic bone marrow transplantation. AB - We report a 40-year-old man who presented with acute onset of hemophagocytic syndrome (HPS) after allogeneic bone marrow transplantation (alloBMT) for acute myelogenous leukemia. On day 8 after alloBMT, the patient suddenly manifested high-grade fever, transfusion-resistant severe anemia, and thrombocytopenia. Neither veno-occlusive disease nor thrombotic microangiopathy was documented. The level of ferritin in serum was elevated to 1192 ng/mL. A bone marrow aspiration test on day 16 showed a markedly increased number of activated macrophages showing massive hemophagocytosis. Serum levels of interferon-gamma, soluble interleukin-2 receptor, interleukin-6, tumor necrosis factor-alpha, and macrophage colony-stimulating factor (M-CSF) were elevated. From these findings, we determined his transfusion-resistant cytopenias to be attributable to HPS. No viruses (including cytomegalovirus, Epstein-Barr virus, human herpes-virus-6, parvovirus B19, and adenovirus B11) were detected in serum or urine by polymerase chain reaction amplification. We speculate that in addition to the administration of M-CSF, hypercytokinemia during the early phase post-alloBMT might have contributed to the onset of HPS in this patient. Methylprednisolone pulse therapy was very effective for the treatment of the HPS. This case reveals that HPS could develop after alloBMT, even when engraftment of hematopoietic cells is not confirmed. PMID- 11039677 TI - No increased risk of thrombosis in heterozygous congenital dysplasminogenemia. AB - To assess the risk of thrombosis in congenital dysplasminogenemia, we studied 10 unrelated families with this disorder. The probands were excluded from the analysis of data to prevent bias in the selection of subjects. Positive thrombotic histories were found in 1 of the 25 family members determined to have heterozygous congenital dysplasminogenemia and in 2 of their 41 biochemically unaffected relatives. The percentages of family members with no history of thrombosis up to a given age among subjects with and without congenital dysplasminogenemia were analyzed by the Kaplan-Meier method. No significant difference between the 2 groups was observed by generalized Wilcoxon test (P = .32) or Cox-Mantel test (P = .62). These findings suggest that heterozygous congenital dysplasminogenemia is not associated with an increased risk of thrombosis. PMID- 11039678 TI - Phenotypic conversion of T-lineage lymphoblasts in the lymph node to myeloblasts in the bone marrow during relapse after allogeneic bone marrow transplantation. PMID- 11039679 TI - Activated protein C resistance in Budd-Chiari syndrome. PMID- 11039680 TI - Observed decline in the rate of death among Japanese hemophiliacs infected with HIV-1. PMID- 11039681 TI - MRI neuroimaging of childhood psychiatric disorders: a selective review. AB - Over the past 10 years, innovations in physics and computer science have promoted magnetic resonance imaging (MRI) as an essential tool for investigating the biological substrates of psychiatric disorders. Requiring no radiation exposure, MRI is now the preferred imaging technique for pediatric populations. However, the rapid technical advances in MRI pulse sequences, data processing, and analysis have made it increasingly complex for clinicians to compare and critically evaluate MRI research studies. This paper selectively reviews MRI research on five psychiatric conditions occurring in childhood or adolescence: ADHD, autism, childhood-onset schizophrenia, Tourette syndrome, and early-onset depression. The selection of papers reviewed was based on four criteria: the originality of the idea underlying the paper, the quality of the sample and methodologies used, the presence of controversial findings in the paper, and whether the paper was a clear illustration of specific methodological strengths or weaknesses. The two goals of this review paper are to update clinicians on morphometric brain imaging in child psychiatry and the methodological issues pertaining to image acquisition and analysis, and to promote critical reading of future MRI studies. PMID- 11039682 TI - From pogroms to "ethnic cleansing": meeting the needs of war affected children. AB - Children are both the direct and indirect targets during wars. They are directly affected by violence aimed at them and their families; they are indirectly affected by the distress caused to their families; they may be internally displaced or find themselves crossing borders as asylum seekers. Their experiences during and immediately after war militate against their developing in a safe, secure, and predictable environment. Their human rights are compromised and their mental health put at risk. Whether in the country at and after war, or in the country that offers refuge, children's mental health needs have to be properly assessed and met. In many cases, children may only require a sense of safety and support via their family and school. In other cases, they require more complicated psychosocial interventions that address the various stress reactions they manifest. This paper addresses these issues against the context of a major community-based programme in Mostar in Bosnia during the recent civil war there. It argues that we have reasonably good screening measures to identify children at high risk of developing mental health problems. It presents an hierarchical model of support and intervention whereby psychosocial help is delivered primarily through schools with only a small proportion of more complex needs being met by specially trained mental health professionals. There is a strong need to evaluate various methods of delivering help and to develop new ways of reaching needy children in a nonstigmatising way. PMID- 11039683 TI - Preconditions and outcome of inpatient treatment in child and adolescent psychiatry. AB - Inpatient care is expensive and should ideally be provided for children and adolescents with the most serious psychiatric disorders. However, only little is known about inpatient treatment, e.g. the factors influencing hospital admission, the content of care in the hospital, the appropriate norms for the duration of inpatient stays, the inpatient arrangements that result in the best outcomes, or connection with necessary aftercare services. There are many methodological problems with existing research. However, it can cautiously be concluded that psychiatric hospitalisation of children and adolescents is often beneficial, particularly if special aspects of treatment are fulfilled (e.g. good therapeutic alliance, treatment with a cognitive-based problem-solving skills training package, or planned discharge) and aftercare services are available. The continuum-of-care model is promising because it provides opportunities to achieve better integration between inpatient interventions and aftercare services. PMID- 11039684 TI - The treatment of childhood social phobia: the effectiveness of a social skills training-based, cognitive-behavioural intervention, with and without parental involvement. AB - Fifty children aged 7-14 years with a principal diagnosis of social phobia were randomly assigned to either child-focused cognitive-behaviour therapy (CBT), CBT plus parent involvement, or a wait list control (WLC). The integrated CBT program involved intensive social skills training combined with graded exposure and cognitive challenging. At posttreatment, significantly fewer children in the treatment conditions retained a clinical diagnosis of social phobia compared to the WLC condition. In comparison to the WLC, children in both CBT interventions showed significantly greater reductions in children's social and general anxiety and a significant increase in parental ratings of child social skills performance. At 12-month follow-up, both treatment groups retained their improvement. There was a trend towards superior results when parents were involved in treatment, but this effect was not statistically significant. PMID- 11039685 TI - Family therapy for adolescent anorexia nervosa: the results of a controlled comparison of two family interventions. AB - This paper reports the results of a randomised treatment trial of two forms of outpatient family intervention for anorexia nervosa. Forty adolescent patients with anorexia nervosa were randomly assigned to "conjoint family therapy" (CFT) or to "separated family therapy" (SFT) using a stratified design controlling for levels of critical comments using the Expressed Emotion index. The design required therapists to undertake both forms of treatment and the distinctiveness of the two therapies was ensured by separate supervisors conducting live supervision of the treatments. Measures were undertaken on admission to the study, at 3 months, at 6 months and at the end of treatment. Considerable improvement in nutritional and psychological state occurred across both treatment groups. On global measure of outcome, the two forms of therapy were associated with equivalent end of treatment results. However, for those patients with high levels of maternal criticism towards the patient, the SFT was shown to be superior to the CFT. When individual status measures were explored, there were further differences between the treatments. Symptomatic change was more marked in the SFT whereas there was considerably more psychological change in the CFT group. There were significant changes in family measures of Expressed Emotion. Critical comments between parents and patient were significantly reduced and that between parents was also diminished. Warmth between parents increased. PMID- 11039686 TI - Effects of early maternal depression on patterns of infant-mother attachment: a meta-analytic investigation. AB - We analysed results from seven American and British studies that compared groups of mothers with and without clinically diagnosed depression, and assessed the attachment category of their infants (under 3 years) using the Strange Situation. The samples were predominantly middle-income and free of risk factors other than maternal depression. Meta-analysis using loglinear modelling and standardised residuals showed that the effect of depression on the distribution of infants' attachment was statistically heterogeneous. However, after removing one outlier study, the effect of depression was homogeneous across the remaining six studies. Infants of depressed mothers showed significantly reduced likelihood of secure (B) attachment and marginally raised likelihood of avoidant (A) and disorganised (D) attachment. The first two effects varied considerably in magnitude between studies, whereas the increase in disorganised attachment, from 17% to 28% on average, was consistent. PMID- 11039687 TI - Maternal depressive symptoms affect infant cognitive development in Barbados. AB - This longitudinal study is part of a series examining the relationships between maternal mood, feeding practices, and infant growth and development during the first 6 months of life in 226 well-nourished mother-infant dyads in Barbados. In this report, we assessed maternal moods (General Adjustment and Morale Scale and Zung Depression and Anxiety Scales), feeding practices (scales describing breast feeding and other practices associated with infant feeding in this setting), and infant cognitive development (Griffiths Mental Development Scales). Multivariate analyses, with and without controlling for background variables, established significant relationships between maternal moods and infant cognitive development. Infants of mothers with mild moderate depression had lower Griffiths scores than infants of mothers without depression. Maternal depressive symptoms and lack of trust at 7 weeks predicted lower infant social and performance scores at 3 months. Maternal moods at 6 months were associated with lower scores in motor development at the same age. Although no independent relationships emerged between feeding practices and infant cognitive development, the combination of diminished infant feeding intensity and maternal depression predicted delays in infant social development. These findings demonstrate the need to carefully monitor maternal moods during the postpartum period, in order to maximize the benefits of breast-feeding and related health programs to infant cognitive development. PMID- 11039688 TI - Insecure and disorganised attachment in children with a pervasive developmental disorder: relationship with social interaction and heart rate. AB - This study on children with a Pervasive Developmental Disorder (PDD; N = 32), children with developmental language disorder (N = 22), and normally developing children (N = 28) sought to answer questions concerning attachment and autistic behaviour. We could replicate the finding that children with a PDD are able to develop secure attachment relationships to their primary caregiver. Children with PDD who had an insecure attachment showed fewer social initiatives and responses than children with PDD who had a secure attachment, even when the insecurely and securely attached PDD children were matched on chronological and mental age. Children with both a PDD and mental retardation were more often classified as disorganised. Three findings suggested that a disorganised attachment does not merely reflect the presence of "autistic" behaviour: (1) children with PDD did not reveal higher rates of a disorganised attachment than matched comparison children; (2) having a PDD diagnosis and having a disorganised attachment were found to be associated with opposite effects on an ethological measure of level of behavioural organisation; and (3) a disorganised attachment but not a PDD diagnosis was associated with an increase in heart rate during parting with the caregiver and a decrease in heart rate during reunion. PMID- 11039689 TI - Atypical interference of local detail on global processing in high-functioning autism and Asperger's disorder. AB - This study explored the claim that individuals with autism and Asperger's disorder tend to process locally rather than holistically. Participants observed a large or "global" number composed of smaller or "local" numbers. The response was contingent upon the identification of either the large stimulus or the small stimuli. Relative to age, sex, and IQ matched controls, global processing in children and adolescents with autism (N = 12) and Asperger's disorder (N = 12) was more vulnerable when the local stimuli were incongruent. The autism group made more global errors than their matched control group, regardless of whether there was local incongruence. In contrast, the Asperger's disorder group made a similar number of global errors as their respective control group. These results were discussed in relation to an "absence of global precedence" notion, "weak central coherence" theory, and right-hemisphere dysfunction. The neurobiological significance of these findings were discussed in the context of a fronto-striatal model of dysfunction. PMID- 11039690 TI - Educational, psychosocial, and sexual outcomes of girls with conduct problems in early adolescence. AB - This paper examines the extent to which conduct problems at age 13 are associated with a range of educational, psychosocial, and sexual outcomes at age 18 in a birth cohort of 488 young women. Significant associations were found between early adolescent conduct problems and later risks of educational failure, juvenile crime, substance abuse, mental health problems, and adverse sexual outcomes by late adolescence. These elevated risks were explained, in part, by social, family, and personal disadvantages associated with adjustment at age 13. In addition, there was evidence of a causal chain process in which early adolescent conduct problems were associated with a series of adolescent risk taking behaviours, including delinquent peer affiliations, early-onset sexual behaviour, substance use, and school problems that were, in turn, associated with increased risks of later adverse outcomes. PMID- 11039691 TI - Predictors of functional impairment in children and adolescents. AB - The goal of this study was to investigate the variables that best predict functional impairment in children and adolescents. Two hundred and eight psychiatric and 129 pediatric children aged 7 to 17 years were assessed with measures of psychopathology, functional impairment, temperament, marital discord, educational style, coping, developmental milestones, stressful life events, medical history, school information, and family history of psychopathology. Multiple regression models adjusted by psychopathology were estimated. The global model, which included all the significant variables in partial models, revealed the following predictors of impairment: receiving review lessons, chronic disease or handicap, the presence of problems the child interpreted as stressful, late onset and long duration of psychopathological problems. These indicators could be useful for the proper identification of children with severe difficulties, in order to provide them with adequate psychological services. PMID- 11039692 TI - Sleep and alertness in children with ADHD. AB - OBJECTIVE: To evaluate sleep and alertness and to investigate the presence of possible underlying sleep/wake disorders in children with attention deficit/hyperactivity disorder (ADHD). METHOD: After 3 nights of adaptation in a room reserved for sleep studies in the department of child psychiatry, children underwent polysomnography (PSG) followed by the Multiple Sleep Latency Test (MSLT) and reaction time tests (RT) during the daytime. Thirty boys diagnosed as having ADHD (DSM-IV), aged between 5 and 10 years, and 22 age- and sex-matched controls participated in the study. All children were medication-free and showed no clinical signs of sleep and alertness problems. RESULTS: No significant differences in sleep variables were found between boys with ADHD and controls. The mean latency period was shorter in children with ADHD. Significant differences were found for MSLT 1, 2 and 3 (p < .05). Mean reaction time was longer in children with ADHD, with significant differences in all tests (p < .05). Number and duration of sleep onsets measured by the MSLT correlated significantly with the hyperactivity-impulsivity and inattentive-passivity indices of the CTRS and CPRS. CONCLUSION: Children with ADHD were more sleepy during the day, as shown by the MSLT, and they had longer reaction times. These differences are not due to alteration in the quality of nocturnal sleep. The number of daytime sleep onsets and the rapidity of sleep-onsets measured as MSLT were found to be pertinent physiological indices to discriminate between ADHD subtypes. These results suggest that children with ADHD have a deficit in alertness. Whether this deficit is primary or not requires further studies. PMID- 11039693 TI - Density-dependent vital rates and their population dynamic consequences. AB - We explore a set of simple, nonlinear, two-stage models that allow us to compare the effects of density dependence on population dynamics among different kinds of life cycles. We characterize the behavior of these models in terms of their equilibria, bifurcations. and nonlinear dynamics, for a wide range of parameters. Our analyses lead to several generalizations about the effects of life history and density dependence on population dynamics. Among these are: (1) iteroparous life histories are more likely to be stable than semelparous life histories; (2) an increase in juvenile survivorship tends to be stabilizing; (3) density dependent adult survival cannot control population growth when reproductive output is high: (4) density-dependent reproduction is more likely to cause chaotic dynamics than density dependence in other vital rates; and (5) changes in development rate have only small effects on bifurcation patterns. PMID- 11039694 TI - Geographical invariance and the strong-migration limit in subdivided populations. AB - Invariance under population subdivision and the strong-migration limit are investigated for digenic samples in neutral models. The monoecious, diploid population is subdivided into a finite number of panmictic colonies that exchange gametes. The backward migration matrix is arbitrary, but time independent and ergodic (i.e., irreducible and aperiodic). Results are derived for the distribution of the place and time of coalescence, for the probability of identity in the model of infinitely many alleles, and for the distribution of the number of nucleotide differences in the model of infinitely many sites without recombination. PMID- 11039696 TI - Nash equilibria for an evolutionary language game. AB - We study an evolutionary language game that describes how signals become associated with meaning. In our context, a language, L, is described by two matrices: the P matrix contains the probabilities that for a speaker certain objects are associated with certain signals, while the Q matrix contains the probabilities that for a listener certain signals are associated with certain objects. We define the payoff in our evolutionary language game as the total amount of information exchanged between two individuals. We give a formal classification of all languages, L(P, Q), describing the conditions for Nash equilibria and evolutionarily stable strategies (ESS). We describe an algorithm for generating all languages that are Nash equilibria. Finally, we show that starting from any random language, there exists an evolutionary trajectory using selection and neutral drift that ends up with a strategy that is a strict Nash equilibrium (or very close to a strict Nash equilibrium). PMID- 11039695 TI - Ring vaccination. AB - Based on the description of an outbreak of foot-and-mouth disease (FMD), a particle model is developed describing the most important properties of this epidemic. Also control measures (mass and ring vaccination) are implemented. This model shows the expected behavior in simulations. Since it is impossible to treat this model analytically, we use ideas of branching processes on two levels to derive a caricature of the particle model. In simulations it is shown that this caricature exhibits similar behavior as the particle system. It is possible to analyze the caricature and, in this way, to obtain expressions for the most important quantities like the reproduction number or the expected final number of infected individuals etc. In this way mass vaccination and ring vaccination can be compared and control strategies can be optimized. PMID- 11039697 TI - Computational model of dot-pattern selective cells. AB - A computational model of a dot-pattern selective neuron is proposed. This type of neuron is found in the inferotemporal cortex of monkeys. It responds strongly to groups of dots and spots of light intensity variation but very weakly or not at all to single dots and spots that are not part of a pattern. This non-linear behaviour is quite different from the spatial frequency filtering behaviour exhibited by other neurons that react to spot-shaped stimuli, such as neurons with centre-surround receptive field profiles found in the lateral geniculate nuclei and layer 4Cbeta of V1. It is implemented in the proposed computational model by using an AND-type non-linearity to combine the responses of centre surround cells. The proposed model is capable of explaining the results of neurophysiological experiments as well as certain psychophysical observations. PMID- 11039698 TI - A first-passage-time analysis of the periodically forced noisy leaky integrate and-fire model. AB - We present a general method for the analysis of the discharge trains of periodically forced noisy leaky integrate-and-fire neuron models. This approach relies on the iterations of a stochastic phase transition operator that generalizes the phase transition function used for the study of periodically forced deterministic oscillators to noisy systems. The kernel of this operator is defined in terms of the the first passage time probability density function of the Ornstein Uhlenbeck process through a suitable threshold. Numerically, it is computed as the solution of a singular integral equation. It is shown that, for the noisy system, quantities such as the phase distribution (cycle histogram), the interspike interval distribution, the autocorrelation function of the intervals, the autocorrelogram and the power spectrum density of the spike train, as well as the input-output cross-correlation and cross-spectral density can all be computed using the stochastic phase transition operator. A detailed description of the numerical implementation of the method, together with examples, is provided. PMID- 11039699 TI - Synchronous oscillation in the cerebral cortex and object coherence: simulation of basic electrophysiological findings. AB - A lumped continuum model for electrocortical activity was used to simulate several established experimental findings of synchronous oscillation which have not all been previously embodied in a single explanatory model. Moving-bar visual stimuli of different extension, stimuli moving in different directions, the impact of non-specific cortical activation upon synchronous oscillation, and the frequency content of EEG associated with synchrony were considered. The magnitude of zero lag synchrony was primarily accounted for by the properties of the eigenmodes of the travelling local field potential superposition waves generated by inputs to the cortex, largely independent of the oscillation properties and associated spectral content. Approximation of the differences in cross correlation observed with differently moving bar stimuli, and of the impact of cortical activation, required added assumptions on (a) spatial coherence of afferent volleys arising from parts of a single stimulus object and (b) the presence of low-amplitude diffuse field noise, with enhancement of cortical signal/noise ratio with respect to the spatially coherent inputs, at higher levels of cortical activation. Synchrony appears to be a ubiquitous property of cortex-like delay networks. Precision in the modelling of synchronous oscillation findings will require detailed description of input pathways, cortical connectivity, cortical stability, and aspects of cortical/subcortical interactions. PMID- 11039700 TI - Dynamic process of information transmission complexity in human brains. AB - Based on a complexity analysis of mutual information transmission of EEG developed by us [Xu J, Liu Z, Liu R, Yang Q (1997) Physica D 106: 363-374], dynamic processes of the complexity of mutual information transmission in human brains were studied. To diminish possible problems due to coarse graining preprocessing, some new measures of complexity were used. The results show that, just before and after generalized seizures, the complexities of almost all information transmission between different brain areas drop significantly; there is also a temporary decrease of complexity when subjects shift their attention. The above facts suggest that there is a transient decrease of information transmission complexity when brain state changes occur suddenly. Mental arithmetic tasks activate the left temporal lobe to exchange more information with other brain areas. The results hint that the methods used here might be an approach to observe quick processes in the living brain. PMID- 11039701 TI - Relevance of nonlinear lumped-parameter models in the analysis of depth-EEG epileptic signals. AB - In the field of epilepsy, the analysis of stereoelectroencephalographic (SEEG, intra-cerebral recording) signals with signal processing methods can help to better identify the epileptogenic zone, the area of the brain responsible for triggering seizures, and to better understand its organization. In order to evaluate these methods and to physiologically interpret the results they provide, we developed a model able to produce EEG signals from "organized" networks of neural populations. Starting from a neurophysiologically relevant model initially proposed by Lopes Da Silva et al. [Lopes da Silva FH, Hoek A, Smith H, Zetterberg LH (1974) Kybernetic 15: 27-37] and recently re-designed by Jansen et al. [Jansen BH, Zouridakis G, Brandt ME (1993) Biol Cybern 68: 275 283] the present study demonstrates that this model can be extended to generate spontaneous EEG signals from multiple coupled neural populations. Model parameters related to excitation, inhibition and coupling are then altered to produce epileptiform EEG signals. Results show that the qualitative behavior of the model is realistic; simulated signals resemble those recorded from different brain structures for both interictal and ictal activities. Possible exploitation of simulations in signal processing is illustrated through one example; statistical couplings between both simulated signals and real SEEG signals are estimated using nonlinear regression. Results are compared and show that, through the model, real SEEG signals can be interpreted with the aid of signal processing methods. PMID- 11039702 TI - Spinal motor control system incorporates an internal model of limb dynamics. AB - The existence and utilization of an internal representation of the controlled object is one of the most important features of the functioning of neural motor control systems. This study demonstrates that this property already exists at the level of the spinal motor control system (SMCS), which is capable of generating motor patterns for reflex rhythmic movements, such as locomotion and scratching, without the aid of the peripheral afferent feedback, but substantially modifies the generated activity in response to peripheral afferent stimuli. The SMCS is presented as an optimal control system whose optimality requires that it incorporate an internal model (IM) of the controlled object's dynamics. A novel functional mechanism for the integration of peripheral sensory signals with the corresponding predictive output from the IM, the summation of information precision (SIP) is proposed. In contrast to other models in which the correction of the internal representation of the controlled object's state is based on the calculation of a mismatch between the internal and external information sources, the SIP mechanism merges the information from these sources in order to optimize the precision of the controlled object's state estimate. It is demonstrated, based on scratching in decerebrate cats as an example of the spinal control of goal-directed movements, that the results of computer modeling agree with the experimental observations related to the SMCS's reactions to phasic and tonic peripheral afferent stimuli. It is also shown that the functional requirements imposed by the mathematical model of the SMCS comply with the current knowledge about the related properties of spinal neuronal circuitry. The crucial role of the spinal presynaptic inhibition mechanism in the neuronal implementation of SIP is elucidated. Important differences between the IM and a state predictor employed for compensating for a neural reflex time delay are discussed. PMID- 11039703 TI - The efficacy of predeposited autologous blood transfusions in general pediatric surgery. AB - Allogeneic blood transfusions are associated with a risk of infection, immunological reactions, immunosuppression, and the induction of antibodies in blood cells. We report our results of giving predeposited autologous blood transfusions (PABT) to children when it was anticipated that transfusions would be required for an elective operation. Autologous blood was collected for deposit from 16 patients ranging in age from 1 to 11 years old (mean 5.6 years old, mode 4 years old), and weighing from 9.7 to 42 kg (mean 20.8kg). They included 12 patients with pectus excavatum (funnel chest) and 4 patients with choledochal cyst (CBD). Blood was collected once from 2 patients and twice from the other 14 patients, then centrifuged and stored in a freezer at -80 degrees C. Between 7 and 14 ml/kg was collected at one time, the total mean volume of predeposited blood being 21.0 +/- 3.3 ml/kg for the children operated on for funnel chest, and 16.2 +/- 4.5 ml/ kg for those operated on for CBD. None of the patients required allogeneic transfusions and no complications occurred. PABT was found to be a safe and effective means for elective general pediatric surgical procedures for avoidance of allogeneic blood transfusion. PMID- 11039704 TI - Increased fibrinolytic activity and body cavity coagula. AB - When a large volume of coagulum remains in the body cavity after trauma or surgery, secondary fibrinolysis occurs, which disturbs the hemostatic balance and results in rebleeding. To better understand this condition, we conducted a clinical study on patients with and without coagula and an experimental study on fibrinolytic activity in a rat model. The results of the clinical study showed that when coagula existed in the body cavity, the blood levels of the fibrin degradation products D-dimer and fibrinopeptide Bbeta15-42 remained high compared with when subjects were under similar stress but without the presence of coagula. In the experimental studies, fibrinolytic activity of the omentum, measured by the fibrin plate method, was higher in rats with hemoperitoneum. This suggests that increased fibrinolytic activity may lead to rebleeding from the area of transient hemostasis when coagulum is present in the body cavity. PMID- 11039705 TI - Prognostic factors in the surgical treatment of ruptured abdominal aortic aneurysms. AB - Although the operative mortality following elective aneurysmectomy has achieved satisfactory results, that following surgery for ruptured abdominal aortic aneurysms (AAAs) remains high. The purpose of this study was to identify the factors affecting the mortality rate associated with the treatment of ruptured AAAs. Between 1978 and 1999, 33 patients underwent emergency surgery for a ruptured AAA. The operative mortality was 33.3% and in-hospital mortality was 6.0%. Hypotension, defined as a systolic blood pressure <80 mmHg, was seen in 19 patients at the time of presentation, 9 of whom underwent surgery in this state. In the remaining 10 patients, it was possible to increase the systolic blood pressure to > or =80 mmHg preoperatively. Of the 11 patients who died within 30 days of surgery, 9 had hypotension at the time of induction of anesthesia and only 2 had a systolic blood pressure of > or =80 mmHg. A satisfactory outcome was achieved in patients whose condition met the following criteria: a systolic blood pressure > or =80 mmHg at the time of operation, minimal aortic cross-clamping time, less blood loss and blood transfusions, and a shorter operation time to repair the ruptured AAA. Concomitant heart disease was also found to be an important prognostic factor. PMID- 11039706 TI - The fate of small aneurysms of the internal iliac artery following proximal ligation in abdominal aortic aneurysm repair. AB - To assess the natural history of small internal iliac artery aneurysms (IIAA) measuring 2.0-3.0 cm in diameter, proximally ligated in association with abdominal aortic aneurysm (AAA) repair, we examined 9 of 12 patients who underwent this type of surgery. Postoperative computed tomography scanning demonstrated that three IIAAs were still patent and the other six were thrombosed. An increase in the maximum diameter from that at the time of surgery was seen in four IIAAs. One patient suffered serious complications in that a dilated IIAA caused right ureteral obstruction and subsequent hydronephrosis accompanied by unilateral renal dysfunction. This was successfully treated by resection of the IIAA. The findings of this analysis led us to conclude that small IIAAs associated with AAA repair should be treated by either endoaneurysmorrhaphy or resection of the aneurysm after both proximal and distal ligation. PMID- 11039707 TI - Does topical cooling alleviate ischemia/reperfusion injury during inflow occlusion in hepatectomy? Results of an experimental and clinical study. AB - This study was undertaken to evaluate whether topical cooling can alleviate ischemia/reperfusion injury, after continuous inflow occlusion during hepatectomy. Using a canine model of 70% partial liver ischemia (60 min), alteration in the subcellular (cytoplasm, mitochondria, nucleus) elements calcium, sodium, potassium, and chloride, and liver functions following reperfusion were compared between control livers and livers subjected to topical cooling down to 23 degrees +/- 4.9 degrees C by seeding ice slush over the ischemic lobe. The elements were determined by X-ray microanalysis using liver biopsy specimens. A similar clinical study was undertaken examining ten patients with hepatocellular carcinoma and chronic liver disease who underwent right-sided segmentectomy under continuous right inflow occlusion, five of whom were given topical cooling and five of whom were not. In the experimental study, postreperfusion worsening of liver function tests was significantly suppressed in the cooling group, which was associated with the suppression of subcellular Ca, Na, and Cl increases and K decreases after reperfusion. In the clinical study, the occlusion time was significantly longer in the hypothermic patients than in the normothermic patients, but no significant differences in postoperative liver function or postischemic increases in Ca, Na, or Cl and decreases in K were observed. These experimental and clinical findings suggest that topical cooling alleviates ischemic insult and enhances safe prolonged inflow occlusion. PMID- 11039708 TI - The differential cardioprotective effects of nucleoside transport inhibitor on moderate and deep hypothermic ischemia with cold cardioplegia. AB - The differences in the cardioprotective effects of nucleoside transport inhibitor (NTI) which is known to accumulate endogenous adenosine, on moderate and deep hypothermic ischemia, were examined. Using the Langendorff model, isolated, perfused rat hearts were arrested with cold cardioplegia and subjected to 90 min of global ischemia followed by 40 min of reperfusion. The temperature during ischemia was maintained at either 10 degrees C (groups 1 and 2) or 25 degrees C (groups 3 and 4). In groups 2 and 4, NTI in the form of R75231, 1 mg/l, was added to the cardioplegic solution. The intramyocardial adenosine triphosphate content at the end of ischemia was significantly lower in the moderate hypothermia groups than in the deep hypothermia groups. In the moderate hypothermia groups, NTI significantly enhanced the adenosine accumulation at the end of ischemia. Moreover, the recovery of both the contractile function and coronary flow rate in group 4 was superior to that in group 3, and was similar to those in groups 1 and 2. The addition of NTI to the cardioplegic solution generated a sufficient cardioprotective effect in moderate hypothermic ischemia, but not in deep hypothermic ischemia. The mechanism of this discrepancy is attributed to the differences in the levels of endogenous adenosine accumulated during ischemia. PMID- 11039709 TI - Video-assisted thoracoscopy system guidance in linear radiofrequency ablation. AB - The transcatheter creation of linear endocardial lesions in the atria has been attempted to restore sinus rhythm in patients with atrial fibrillation (AF). However, due to fluoroscopic limitations, there are a number of technical difficulties involved with using this procedure, which include determining the ablation site, orienting the catheter tip, and confirming tip-tissue contact. The present study was performed to assess the feasibility of employing a transthoracic approach to linear radiofrequency ablation using a video-assisted thoracoscopy system (VATS) to anatomically guide the experimental setting in beating swine hearts. AF was induced pharmacologically by aconitine solution. Epicardial radiofrequency linear ablation of the right atrium was conducted under VATS monitoring using an ablation catheter that was inserted and manipulated through trocar ports. The ablation energy setting was 80 degrees C with 30s duration per ablation. The thoracoscopic visual field for transthoracic ablation was adequate, and the ablation catheter was positioned and anchored safely on the atrial epicardium. The restoration of sinus rhythm was confirmed in the limb lead and atrial electrograms, and transmural heat degeneration was confirmed by postmortem histological examination in all specimens. Our results suggest the potential usefulness of VATS for providing adequate anatomical guidance in epicardial linear radiofrequency ablation. PMID- 11039710 TI - Human skin xenograft rejection in CD45 exon-6 knockout mice: the implication of involvement of a direct pathway. AB - The results of previous studies indicate that only CD4+ T cells generated via the indirect pathway play an essential role in causing discordant skin xenograft rejection. The present study was conducted in an attempt to clarify further the roles of effector T cells generated via direct pathways on discordant xenograft rejection using CD45 exon-6 knockout (CD45-/-; C57BL/6 (B6): H-2b) mice. It has been strongly suggested that CD45 exon-6 knockout mice have profound impairment in T-cell functions via an indirect pathway. When human skin was grafted onto untreated normal C57BL/6 (B6; H-2b) mice, rejection occurred within 12 days; however, in the CD45 exon-6 knockout mice, the grafts lasted for slightly longer as in fully allogeneic C3H (H-2k) skin rejection, with a mean survival time +/- SD of 19.4 +/- 1.5 days and median survival times of 19 days. The difference in survival periods between the human and C3H skin grafts in the CD45 knockout mice was not statistically significant. Both CD4+ and CD8+ T cells seemed activated in the spleens of these CD45 exon-6 knockout mice 10 days after the human skin grafting. These results suggest that effector T cells generated via a direct pathway can cause discordant skin xenograft rejection, and that CD45 exon-6 knockout mice can generate effector T cells via a direct pathway to reject discordant skin xenografts, similarly to fully allogeneic skin allografts. PMID- 11039711 TI - Humoral human xenoreactivity against isolated pig pancreatic islets. AB - It is widely believed that the hyperacute rejection of vascularized xenografts in the pig-to-human combination is triggered by the binding of human preformed natural antibodies (PNAbs) to the Galalpha.(1,3)Gal epitope in pig endothelium and the subsequent activation of complement. However, it remains poorly defined whether xenogeneic pig pancreatic islets are damaged by antibody and complement mediated mechanisms. We examined the expression of Galalpha(1,3)Gal on isolated adult pig islets and the presence of PNAbs in normal human sera directed against islets, using immunofluorescence staining and confocal laser scanning microscopy. The pig islets were not stained with Galalpha(1,3)Gal-specific lectin GSIB4, however, the exocrine cells reacted strongly with GSIB4, indicating that the Galalpha(1,3)Gal epitope was highly expressed on exocrine cells, but not on islets. Human sera showed weak reactivity of IgM and IgG class PNAbs to the islets, but strong reactivity to the exocrine cells. Furthermore, we investigated the cytotoxic effect of human serum on pig islets using an in vitro model of pig to-human islet transplantation. The incubation of pig islets with normal human sera for 45 min resulted in less than 10% specific lysis despite the binding of PNAbs, whereas exposure of porcine aortic endothelial cells to the same human sera caused 56% complement-mediated lysis, determined using a MTT cytotoxic assay. These results support the view that pig islets might not undergo early antibody and complement-mediated rejection in humans. PMID- 11039712 TI - Solitary brain metastasis from papillary thyroid carcinoma in a patient with depression: report of a case. AB - Papillary carcinoma of the thyroid is a common thyroid malignancy with a relatively good prognosis. However, distant metastases may develop and become threatening, particularly to older patients, in a more aggressive manner. We report herein the clinical, radiological, and pathological findings of a patient with papillary thyroid carcinoma who had a solitary cerebral metastasis. The patient had been suffering from depression and had already undergone a hemithyroidectomy for primary thyroid carcinoma, and was known to have metastatic thyroid carcinoma of the lungs and bone. After the removal of the remnant thyroid gland prior to radioiodine (131I) therapy, he developed additional problems related to depression. Electroencephalography played an important role in identifying suspected brain metastasis and computed tomography demonstrated a space-occupying lesion in the left cerebral hemisphere. Consequently, an early removal of intracranial mass could be performed without any further life threatening complications. Moreover, after removal of the brain mass the patient's depression improved immediately without the use of any antidepressants. This case report indicates the possibility that a patient's depression might be associated with brain metastasis from papillary thyroid carcinoma, and also suggests that an early diagnosis with the appropriate surgical management of a brain metastasis followed by radioiodine therapy could be valuable for achieving a prolonged disease-free period. PMID- 11039713 TI - Undifferentiated spindle-cell sarcoma of the chest wall with vascular endothelial growth factor expression: report of a case. AB - The expressions of some growth factors have been immunohistochemically confirmed in several kinds of tumors, and in particular the expression of vascular endothelial growth factor (VEGF) has been reported to be closely related to tumor cell proliferation. We report herein a case of undifferentiated spindle-cell sarcoma arising in the chest wall with VEGF expression. A 67-year-old man, who presented with coughing, was found to have an abnormal shadow on his right chest wall. He was admitted to Chiba Rosai Hospital and preoperative diagnosis of the tumor was sarcoma. The tumor was thus resected along with the right chest wall and right lower lobe of the lung. Histopathologically, the tumor cells were spindle-shaped and showed severe atypism. The tumor cells were positive for vimentin and VEGF antibody with immunohistochemical staining, but they did not show differentiation to any special type of sarcoma. The tumor was diagnosed to be undifferentiated spindle-cell sarcoma. The microvessel density of the tumor was measured using CD34 and it was found to be higher than the average density of usual sarcomas. The prognosis of this case was poor. The patient died of tumor metastasis to the lung and bone 1 year later in spite of the fact that the tumor was resected. PMID- 11039714 TI - An uncommon case of simultaneous malignant tumors: bronchioloalveolar cancer and Ewing's sarcoma in a 17-year-old girl: report of a case. AB - This report describes a 17-year-old girl with a simultaneous occurrence of bronchioloalveolar cancer and Ewing's sarcoma of the femur, which is a previously unreported association. This case emphasizes the existence of multiple primary malignant neoplasms even in adolescents. Primary lung cancer should therefore be considered in patients under 19 years of age who present with abnormal pulmonary lesions. PMID- 11039715 TI - Rapidly growing primary cardiac leiomyosarcoma: report of a case. AB - Primary cardiac leiomyosarcomas are very rare. A 19-year-old man was admitted to a local hospital with dyspnea and hemoptysis. He was later transferred to our hospital because of his worsening dyspnea. An enhanced chest computed tomography scan demonstrated a large mass in the left atrium. A transthoracic echocardiogram showed a large mobile mass in the left atrium. The tumor was totally resected. The pathohistological examination showed leiomyosarcoma. The tumor rapidly recurred. and a second and third operation were performed. After the third operation, the patient was treated with radiotherapy. There was no local recurrence but multiple distant metastases were found 2 months after completion of radiation therapy. PMID- 11039716 TI - Effect of liver transplantation in a twin for biliary atresia on physical development and intellectual performance: report of a case. AB - A case of twins, one of whom suffered from biliary atresia, is described herein. Although the patient had been doing well until 11 years of age after previously undergoing a primary hepatic portoenterostomy, she had to then undergo a liver transplantation due to severe refractory cholangitis at 14 years of age. Although the patient's intellectual performance had severely declined due to the progression of her illness for several years, it completely recovered after the liver transplantation. In cases where the physical development in childhood has been well preserved, liver transplantation might therefore offer the chance for a full recovery of deteriorated intellectual performance. PMID- 11039717 TI - Portal vein thrombosis caused by microwave coagulation therapy for hepatocellular carcinoma: report of a case. AB - Microwave coagulation therapy (MCT) is one of the treatment modalities for patients with hepatocellular carcinoma (HCC). A 67-year-old man with liver cirrhosis underwent MCT during a laparotomy for a deeply located HCC (2.5 cm in diameter) at the border of the anterior and posterior segments of the right hepatic lobe. Two weeks after MCT, he complained of abdominal fullness. Portal vein thrombosis (PVT) was diagnosed because he had massive ascites and an echogenic mass in the portal vein on abdominal ultrasonography. PVT was successfully treated by fibrinolytic therapy with a selective infusion of urokinase via the superior mesenteric artery (SMA). There have been few reports on PVT as a complication of MCT. Attention should be paid to the possible occurrence of PVT as a critical complication after MCT for liver tumors adjacent to the portal vein. Fibrinolytic therapy via the SMA is thus considered to be an effective approach for PVT after MCT. PMID- 11039718 TI - Hepatic sclerosing hemangioma mimicking a metastatic liver tumor: report of a case. AB - We present herein the case of a sclerosing hemangioma of the liver which was extremely difficult to differentiate from liver metastasis of rectal cancer, in a 67-year-old woman. All the radiological findings were compatible with liver metastasis; however, marginal pooling of the tumor revealed by computed tomographic angiography and magnetic resonance imaging scans was inconsistent with a diagnosis of liver metastasis. At laparotomy, the tumor was macroscopically unusual in that it was yellowish elastic-hard with a very clear margin, and thus, it did not have the appearance of a metastatic tumor. Mile's operation and a partial hepatectomy were performed, followed by an uneventful postoperative course and no signs of recurrence. The carcinoembryonic antigen (CEA) level in the peripheral blood was not elevated at any time. The postoperative pathological diagnosis was a rare hepatic tumor, namely, a "sclerosing hemangioma," based on the findings of cellular fibrous stroma containing vascular channels with flattened endothelial cells. Preoperatively differentiating between sclerosing hemangioma and a metastatic liver tumor from colorectal cancer may be very difficult; however, this case demonstrated some interesting characteristics, namely, the serum CEA level was not elevated, marginal pooling of the tumor was found in the enhanced radiological findings, and the tumor was macroscopically unusual. Therefore, the possibility of sclerosing hemangioma should be borne in mind when considering the differential diagnosis of patients suspected of having colorectal liver metastasis. A preoperative biopsy should be carried out and when a laparotomy is performed under the misdiagnosis of colorectal liver metastasis, it is advisable that either an intraoperative needle biopsy or a frozen histological analysis be undertaken to avoid unnecessary extended hepatic resection of this rare benign hepatic tumor. PMID- 11039719 TI - Late-type recurrence at the port site of unexpected gallbladder carcinoma after a laparoscopic cholecystectomy: report of a case. AB - A 73-year-old woman had a laparoscopic cholecystectomy for unexpected gallbladder cancer and 9 days later underwent both a liver bed resection and lymph node dissection. Four years later, she underwent a further resection of a port site recurrence of gallbladder cancer and no other site of recurrence was observed. The seeding of cancer cells during the removal of the resected gallbladder might have caused this tumor. This case may show that the port site recurrence did not necessarily indicate an incurable stage of the disease. In addition, an excision of the recurrent tumor also appeared to eliminate the disease in the patient. PMID- 11039720 TI - Lymphoepithelial cyst of the pancreas: report of a case. AB - An extremely rare case of a lymphoepithelial cyst (LEC) of the pancreas is described herein. A pancreatic cystic tumor was initially detected in a 50-year old man at a medical checkup. On admission, his serum carbohydrate antigen (CA) 19-9 level was 8100 U/ml and a computed tomography scan revealed a well circumscribed multilocular cystic tumor in the pancreatic head and body. Magnetic resonance cholangiopancreatography showed no communication between the pancreatic ducts and the tumor. A distal pancreatectomy with lymph node dissection was performed because the lesion was suspected to be a mucinous cystadenoma or cystadenocarcinoma of the pancreas. However, histological examination revealed that the cyst was lined by stratified squamous epithelium and surrounded by lymphoid tissue. thereby confirming the diagnosis of LEC of the pancreas. The superficial layer of squamous epithelium and the cystic contents were found to be immunohistologically positive for CA19-9. Establishing a preoperative diagnosis of LEC is quite difficult because it resembles other cystic neoplasms of the pancreas in radiographic features and is frequently associated with an elevation of serum tumor markers such as CA19-9. PMID- 11039721 TI - A child with adrenocortical adenoma accompanied by congenital hemihypertrophy: report of a case. AB - We report herein the findings of a 7-year-old male child with a ruptured adrenocortical adenoma and congenital hemihypertrophy which was incidentally detected after suffering a trauma. A review of 21 pediatric cases of adrenocortical neoplasms in the literature was made. The patient showed precocious puberty such as pubis and advanced bone age, but an endocrinological examination revealed no definite abnormalities. The right adrenal tumor with hematoma was resected after these evaluations. Adrenocortical adenoma is considered to occur more frequently in female children. However, the incidence of adrenocortical tumors accompanied by congenital hemihypertrophy does not differ between males and females. The outcomes were relatively good, although the observation periods were short in some patients. A large number of patients presented with a tumor and hemihypertrophy on the same side. This finding is of interest when considering the possible association between hemihypertrophy of the organs and tumor proliferation. However, their association in terms of development was unclear. It is necessary for patients with hemihypertrophy to have regular examinations for the possible development of malignant tumors, especially in the kidney, adrenal gland, and liver. PMID- 11039722 TI - Successful angioplasty of the left pulmonary artery using the vascular clip system. AB - Pulmonary angioplasty was successfully performed in two patients using the vascular clip system (VCS). The portion of the pulmonary artery which was directly invaded by primary lung cancer was partially resected. The VCS was used to quickly and easily repair the vascular defect. No bleeding was noted from the clipped vascular walls. Both patients had uneventful recoveries. PMID- 11039723 TI - Selective increase of Nurr1 mRNA expression in mesencephalic dopaminergic neurons of D2 dopamine receptor-deficient mice. AB - The orphan nuclear receptor Nurr1 is critical for the survival of mesencephalic dopaminergic precursor neurons. Little is known about the mechanisms that regulate Nurr1 expression in vivo. Other members of this receptor family have been shown to be activated by dopamine. We sought to determine if Nurr1 expression is also regulated by endogenous dopamine through dopamine receptors. Consequently, we investigated the expression of Nurr1 mRNA in genetically modified mice lacking both functional copies of the D2 dopamine receptor gene and in their congenic siblings. Quantitative in situ hybridization demonstrated a significant increased expression of Nurr1 mRNA in the substantia nigra pars compacta and the ventral tegmental area of D2 dopamine receptor -/- mice. No change in Nurr1 expression was detected in other brain regions, such as the habenular nuclei and temporal cortex. Among the cell groups studied, mesencephalic dopaminergic neurons are unique in that they express both Nurr1 and the D2 dopamine receptor, and synthesize dopamine. Thus, it seems plausible that the selective increase in Nurr1 expression observed in D2 receptor-deficient mice is the consequence of an impaired dopamine autoreceptor function. PMID- 11039724 TI - Effects of transient global ischemia and kainate on poly(ADP-ribose) polymerase (PARP) gene expression and proteolytic cleavage in gerbil and rat brains. AB - Poly (ADP-ribose) polymerase (PARP) is involved in various cellular functions, including DNA repair, the cell cycle and cell death. While PARP activation could play a critical role in repairing ischemic brain damage, PARP inactivation caused by caspase 3-cleavage may also be important for apoptotic execution. In this study we investigated the effects of transient global ischemia and kainic acid (KA) neurotoxicity, in gerbil and rat brains, respectively, on PARP gene expression and protein cleavage. PARP mRNA increased in the dentate gyrus of gerbil brains 4 h after 10 min of global ischemia, which returned to basal levels 8 h after ischemia. KA injection (10 mg/kg) also induced a marked elevation in PARP mRNA level selectively in the dentate gyrus of rat brains 1 h following the injection, which returned to basal levels 4 h after the injection. These observations provide the first evidence of altered PARP gene expression in brains subjected to ischemic and excitotoxic insults. Using both monoclonal and polyclonal antibodies to PARP cleavage products, little evidence of significant PARP cleavage was found in gerbil brains within the first 3 days after 10 min of global ischemia. In addition, there was little evidence of significant PARP cleavage in rat brains within 2 days after kainate (KA) injection. Though these findings show that caspase induced PARP cleavage is not substantially activated by global ischemia and excitotoxicity in whole brain, the PARP mRNA induction could suggest a role for PARP in repairing DNA following brain injury. PMID- 11039725 TI - Sensory stimulation increases cortical expression of the fragile X mental retardation protein in vivo. AB - Fragile X syndrome is a common cause of mental retardation that results from the absence of the fragile X mental retardation protein (FMRP), an RNA binding protein whose function remains unclear. Recent in vitro work has demonstrated that the protein is translated near the synapse in an activity dependent manner [33]. We therefore asked whether expression of FMRP might be altered by neuronal activity in vivo. Using immunoblots of different sub-cellular fractions of the rat somatosensory cortex, we show that the levels of FMRP increase significantly following unilateral whisker stimulation, a model of experience dependent plasticity. This increase is greatest between 2 and 8 h after the stimulus and is seen in both a synaptosomal fraction as well as a sub-cellular fraction enriched for polyribosomal complexes. In contrast, detectable levels of FMRP within the somatosensory cortex show either a decrease or no change after a kainic acid induced seizure compared to water treated controls. Our findings demonstrate that FMRP expression levels are modulated in vivo in response to neuronal activity and suggest a role for FMRP in activity dependent plasticity. PMID- 11039726 TI - Alternative splicing of Drosophila calcium/calmodulin-dependent protein kinase II regulates substrate specificity and activation. AB - Drosophila calcium/calmodulin-dependent protein kinase II is alternatively spliced to generate multiple isoforms that vary only in a region between the calmodulin-binding domain and the association domain. This variation has been shown to modulate activation of the enzyme by calmodulin. In this study we examine the ability of seven of the Drosophila isoforms to phosphorylate purified protein substrates and to be inhibited by a substrate analog, and the response of six of the isoforms to a mutant form of calmodulin (V91G) that was isolated in a genetic screen. Significant variation in Kms for Eag, a potassium channel, and Adf-1, a transcription factor, were found. In the case of the a peptide inhibitor, AC3I, there were significant variations in Ki between isoforms. Kact for V91G calmodulin was increased for all of the isoforms. In addition, one isoform, RI, exhibited a lower Vmax when assayed with this mutant CaM. These results indicate that the variable domain of calcium/calmodulin-dependent protein kinase II is capable of altering the substrate specificity of the catalytic domain and the activation response to calmodulin. PMID- 11039727 TI - Central administration of an antisense oligodeoxynucleotide against type I pituitary adenylate cyclase-activating polypeptide receptor suppresses synthetic activities of LHRH-LH axis during the pubertal process. AB - Central administration of an antisense oligodeoxynucleotide against type I pituitary adenylate cyclase-activating polypeptide receptor suppresses synthetic activities of LHRH-LH axis during the pubertal process In the present study, we determined the expression of pituitary adenylate cyclase-activating polypeptide (PACAP) and PACAP receptor type I (PAC1) genes during juvenile development and the pubertal process. Female rats were assigned--based on uterine weights, the presence and abundance of uterine fluid, and their vaginal patency--to one of the following: anestrus (AE), early proestrus (EP), late proestrus (LP) or first estrus (E). The hypothalami from 22-, 24- and 26-day-old animals and from those in the peripubertal phases of AE, EP, LP and E were collected, and the content of PACAP and PAC1 mRNA was assessed. These levels were found to decrease in EP and LP. To determine the effect of PACAP on prepubertal luteinizing hormone-releasing hormone (LHRH) and LH synthesis through PAC1, a PAC1 antisense oligodeoxynucleotide (ODN) was i.c.v.-administered, and mRNA levels of LHRH, LH beta, and LHRH receptor (LHRH-R) were determined. Prepubertal increases in LHRH, LH beta, and LHRH-R mRNA levels were markedly suppressed, and the onset of puberty was delayed by the i.c.v. injection of the antisense PAC1 ODN. These data suggest that PACAP may play a role in the regulation of hypothalamic LHRH neurons, through which it regulates synthetic machinery of pituitary LH, during the pubertal process. PMID- 11039728 TI - Mu opioid receptor mRNA expression in neuronal nitric oxide synthase immunopositive preoptic area neurons. AB - Nitric oxide (NO) as well as beta-endorphin are involved in the neuroendocrine control of gonadotropin-releasing hormone (GnRH) secretion. Recently, morphological and microdialysis experiments have suggested that beta-endorphin may exert an inhibitory influence on NO release in the preoptic area of rat hypothalamus. The present study determines if the mu opioid receptor mRNA is expressed in neuronal NO synthase (nNOS)-immunopositive neurons and if this expression varies among the regions of the basal forebrain being examined. We found, through the use of immunohistochemical and in situ hybridization techniques, that the mu opioid receptor mRNA is expressed in a representative subpopulation of nNOS-immunoreactive neurons in the rat preoptic area. Interestingly, the mu opioid receptor mRNA/nNOS-immunoreactive coexpression is predominant in the rostral and median preoptic area, containing most of GnRH cell bodies. These results strongly suggest that beta-endorphin, via an action through mu opioid receptors, may directly participate in the regulation of NO production in the preoptic area. Our results strengthen the hypothesis that beta-endorphin may participate in GnRH neuronal modulation at the cell body level by regulating NO release from the interneurons of the preoptic area that express nNOS. PMID- 11039729 TI - The anti-dementia drug nefiracetam facilitates hippocampal synaptic transmission by functionally targeting presynaptic nicotinic ACh receptors. AB - Nefiracetam, a pyrrolidone derivative developed as an anti-dementia drug, persistently potentiated currents through neuronal nicotinic acetylcholine (ACh) receptors (alpha7, alpha4beta2) expressed in Xenopus oocytes, and the potentiation was blocked by either the selective protein kinase C (PKC) inhibitors, GF109203X and staurosporine, or co-expressed active PKC inhibitor peptide. In primary cultures of rat hippocampal neurons, nefiracetam increased the rate of nicotine-sensitive miniature excitatory postsynaptic currents, without affecting the amplitude, and the increase was inhibited by GF109203X. In addition, the drug caused a marked increase in the glutamate release from electrically stimulated guinea pig hippocampal slices, and the effect was abolished by the nicotinic ACh receptor antagonists, alpha-bungarotoxin and mecamylamine. Nefiracetam induced a long-lasting facilitation of synaptic transmission in both the CA1 area and the dentate gyrus of rat hippocampal slices, and the facilitation was inhibited by alpha-bungarotoxin and mecamylamine. Such facilitatory action was still found in the hippocampus with selective cholinergic denervation. The results of the present study, thus, suggest that nefiracetam enhances activity of nicotinic ACh receptors by interacting with a PKC pathway, thereby increasing glutamate release from presynaptic terminals, and then leading to a sustained facilitation of hippocampal neurotransmission. This may represent a cellular mechanism underlying the cognition-enhancing action of nefiracetam. The results also provide the possibility that nefiracetam could be developed as a promising therapeutic drug for senile dementia or Alzheimer's disease. PMID- 11039730 TI - The splice variant D3nf reduces ligand binding to the D3 dopamine receptor: evidence for heterooligomerization. AB - The D3 dopamine receptor belongs to the D2-like family of dopamine receptors. As with other members of this group, the D3 dopamine receptor gene contains introns which allow for alternative splicing of gene products. The best characterized of the human D3 dopamine receptor mRNA splice variants encodes a truncated protein called D3nf. The D3 dopamine receptor and D3nf were epitope-tagged and expressed in Sf9 insect cells by recombinant baculovirus infection. The D3 dopamine receptor showed saturable, high affinity binding of agonists and antagonists, consistent with reported D3 dopamine receptor pharmacology. When the D3 dopamine receptor and D3nf were co-expressed, the apparent density of D3 dopamine receptor expression, as determined by radioligand binding, was significantly lowered compared to D3 dopamine receptor expressed alone. This effect of D3nf was specific for the D3 dopamine receptor, since co-expression with the D2 dopamine receptor or beta2-adrenoceptor had no effect on binding. Confocal immunofluorescence studies were used to confirm that both D3 dopamine receptor and D3nf were well expressed on the cell surface and densitometric analysis of cell surface membrane protein confirmed that D3nf did not significantly alter the amount of D3 dopamine receptor expressed. Photoaffinity labelling with [125I]azidonemonapride showed that the amount of ligand bound by membranes co expressing D3 dopamine receptor and D3nf was significantly less than that bound by membranes expressing D3 dopamine receptor alone. The greatest decrease in binding was observed in the D3 dopamine receptor oligomeric forms. Ligand binding to dimers and tetramers was reduced by 69 and 46%, respectively, indicating effects of a protein-protein interaction. Co-immunoprecipitation confirmed that the D3DR and D3nf interact with each other. These data indicate that D3nf heterodimerizes with the D3 dopamine receptor and decreases the capacity of D3 dopamine receptor to bind ligand. PMID- 11039731 TI - Sleep and wakefulness in c-fos and fos B gene knockout mice. AB - G-protein coupled receptor (GPCR) stimulation has been implicated in the regulation of sleep. Upon stimulation of a GPCR an intracellular cascade involving second and third messengers is initiated. The latter include the fos family of immediate early genes (IEGs). Although there is considerable evidence indicating that IEGs are expressed in response to sleep, the effects of their deletion on sleep is not known. The present study examined sleep-wakefulness in mice lacking the c-fos or fos B genes. Null c-fos mice compared to their wildtype (WT) and heterozygote (het) siblings had more wakefulness and less slow wave sleep (SWS); REM sleep was not affected. The null c-fos mice also had increased delta activity (0.3-4 Hz). In contrast, the null and heterozygote fos B mice had less REM sleep, but the time spent in SWS or wakefulness was not different from their wild-type (WT) siblings. In the null c-fos mice, the increased wakefulness and the reduction in SWS could not be due to a systemic alteration in temperature since the core temperature was similar in all mice. By demonstrating that these IEGs are involved in sleep, we suggest that the deletion of specific genes, even within a family of genes, can have a specific effect on sleep. PMID- 11039732 TI - Studies on the interaction of REST4 with the cholinergic repressor element 1/neuron restrictive silencer element. AB - REST4 is a neuron specific truncated form of the transcription factor REST/NRSE derived by alternative splicing. REST4 was previously shown to block the repressor activity of REST/NRSF by forming a hetero-oligomer, Shimojo et al. [Mol. Cell. Biol. 19 (1999) 6788-6795]. A series of deletion mutants have now been used to characterize REST4 in terms of its structure and DNA binding. REST4 was found to be O-glycosylated between between residues 87 and 152. Binding of REST4 to the cholinergic RE-1/NRSE was approximately 1/10 to 1/20 as strong as full length REST/NRSF. DNA binding was enhanced by deletion of the first 86 residues and was found to require all four of the C-terminal zinc fingers as well as a twelve amino acid sequence preceding the first of these zinc fingers. REST4 can form homo-oligomers, however only the monomer was found to bind to DNA. REST4 binds to the 3' sequence of the cholinergic NRSE suggesting an anti-parallel orientation of the protein to the DNA. PMID- 11039733 TI - Localization of the tandem pore domain K+ channel TASK-1 in the rat central nervous system. AB - Recently, a new family of potassium channels with two pore domains in tandem and four transmembrane segments has been identified. Seven functional mammalian channels have been reported at this time. These channels give rise to baseline potassium currents because they are not gated by voltage and exhibit spontaneous activity at all membrane potentials. Although the physiological role of these ion channels has yet to be determined, three mammalian members of this family (TREK 1, TASK-1, TASK-2) are activated by volatile anesthetics and may therefore contribute to the central nervous system (CNS) depression produced by volatile anesthetics. In this study we used northern blot analysis and immunohistochemical localization to determine the expression of TASK-1 subunits in the CNS. TASK-1 immunoreactivity was prominently found in astrocytes of the hippocampus, in the median eminence, in the choroid plexus, and the granular layer, Purkinje cell layer, and molecular layer of the cerebellum. In the spinal cord, strong TASK-I immunoreactivity was seen in ependymal cells lining the central canal and in white matter. These findings suggest a role for the TASK-1 channel in the production of cerebrospinal fluid and function of hypothalamic neurosecretory cells. PMID- 11039734 TI - Uveoscleral outflow: the other outflow pathway. PMID- 11039735 TI - Screening performance of functional and structural measurements of neural damage in open-angle glaucoma: a case-control study from the Baltimore Eye Survey. AB - PURPOSE: To compare the sensitivity and specificity of four approaches to glaucoma screening. METHODS: Case patients were persons with possible, probable, or definite glaucomatous optic nerve damage, as judged by a glaucoma specialist using Humphrey 24-2 threshold findings and clinical assessment of disc and nerve fiber layer, identified in the population-based Baltimore Eye Survey Follow-up Study. Control patients were participants in the same study, frequency-matched for age, without evidence of glaucomatous optic nerve damage. Participants underwent optic disc photography (Topcon ImageNet), disc imaging (GlaucomaScope), scanning laser polarimetry (Nerve Fiber Analyzer), and suprathreshold field testing (Dicon). RESULTS: A total of 100 case patients with open-angle glaucoma and 149 control patients were included. Objective imaging had the best screening performance. For the GlaucomaScope, a criterion of cup-to-disc ratio of -0.68 had a sensitivity of 72% and specificity of 82% for detecting eyes with definite or probable glaucomatous optic nerve damage. For the nerve fiber layer, a criterion of The Number as > or = 20 had a sensitivity of 69% and specificity of 77% for detecting eyes with definite or probable glaucomatous optic nerve damage. Usable data could be obtained in 93% of participants with the Dicon and the Nerve Fiber Analyzer and in 82% and 87% of participants with the GlaucomaScope and Topcon instruments, respectively. CONCLUSIONS: Vertical cup-to-disc ratio, as measured by the GlaucomaScope or Topcon instruments, and the Nerve Fiber Layer neural network Number had the best combination of sensitivity and specificity among the instruments tested. The Nerve Fiber Analyzer had the highest percentage of participants with usable data. PMID- 11039736 TI - Influence of optic disc size on neuroretinal rim shape in healthy eyes. AB - PURPOSE: To evaluate the influence of optic disc size on segmental neuroretinal rim area in healthy eyes. PATIENTS AND METHODS: The study included 193 eyes of 193 healthy patients with physiologic disc cupping. On 15 degrees color stereophotographic optic disc diapositives, optic disc area and neuroretinal rim area were morphometrically determined in 36 radial optic disc segments each measuring 10 degrees. RESULTS: The correlations of segmental rim area to disc area were significantly strongest (P < 0.01) and the regression lines were steepest in the inferior disc region, and the values were lowest in the temporal disc region. Complementary to the rim data, the correlations of segmental cup area to disc area were significantly strongest (P < 0.01) and the regression lines were steepest in the temporal disc region, and the values were lowest in the inferior disc region. In comparison with neuroretinal rim area, cup area was significantly (P < 0.01) more strongly correlated with disc area and the regression line was steeper in the whole optic disc and in each disc segment. The regional distribution of the widest rim part and smallest rim part was independent of disc size. CONCLUSIONS: The increase of rim area and cup area with increasing disc size differs between various disc regions. Because cup area increases more than rim area with increasing disc size, correction for disc size may be more important for segmental cup area than for segmental rim area. The rim shape with respect to the location of the smallest or broadest rim part is independent of disc size. PMID- 11039737 TI - Reproducibility of measurements with the nerve fiber analyzer (NfA/GDx). AB - PURPOSE: To determine the reproducibility of measurements with the Nerve Fiber Analyzer, a scanning laser polarimeter designed for quantifying glaucoma in healthy patients and patients with glaucoma. The authors also assessed the variance of measurements between instruments. METHODS: Measurements were made with the third generation Nerve Fiber Analyzer, the GDx. The study consisted of three parts. In the first part, the authors measured the right eyes of 10 healthy volunteers on 5 consecutive days. In the second part, 45 patients with glaucoma underwent Nerve Fiber Analyzer measurements of one randomly selected eye on two separate days in a 5-week period. For all 14 available parameters, reproducibility of measurements was expressed in terms of 95% limits of agreement and as the intraclass correlation coefficient. The Nerve Fiber Analyzer software has an option of creating a mean image from a selection of single images; for both parts of the study, the reproducibility of measurements was calculated for a "single image," and a "mean-of-three" image. In the third part of the study, 17 volunteers underwent repeated Nerve Fiber Analyzer measurement sessions on each of three different instruments. Using multivariate analysis of variance, the authors determined the variance of measurements between instruments. RESULTS: The reproducibility of measurements varied considerably across parameters. Limits of agreement in mean images for superior maximum and inferior maximum were 7.2 microm and 7.7 microm, respectively in the healthy volunteers, and 8.7 microm and 7.9 microm, respectively in the patients with glaucoma. For healthy patients, the intraclass correlation coefficient was greater than 90% in 10 of 14 parameters. In patients with glaucoma, the intraclass correlation coefficient was greater than 90% in 13 of 14 parameters. Some parameters reproduced better in a mean than in a single image; these differences, however, were small and generally not statistically significant. The between-instruments component also varied across parameters and was highest in ratio-based parameters. CONCLUSIONS: The reproducibility of measurements varied across parameters. In general, the reproducibility of measurements with the Nerve Fiber Analyzer was high. The reproducibility of measurements was similar between healthy patients and patients with glaucoma. Any measured change in nerve fiber layer thickness would be statistically significant if it exceeded approximately 7 or 8 microm in the superior maximum or inferior maximum parameter in healthy patients. Reproducibility of measurements hardly differed between single images and mean images. The reproducibility of measurements among the three instruments we used was highest for straight parameters. PMID- 11039738 TI - Changes in the nerve fiber layer thickness following a reduction of intraocular pressure after trabeculectomy. AB - PURPOSE: To assess changes in the nerve fiber layer thickness after trabeculectomy using scanning laser polarimetry. METHODS: The authors prospectively enrolled 46 eyes from 46 patients with primary open-angle glaucoma in whom intraocular pressure had been reduced by more than 30% after trabeculectomy without significant ophthalmic complications and from whom good quality images were obtained by a scanning laser polarimetry preoperatively and at 3 to 6 months after trabeculectomy. In each enrolled eye, changes in the nerve fiber layer thickness after surgery in the defined ring (1.8 disc diameters) around the optic disc were calculated in 10 degrees intervals (36 sectors in total) and in the following 4 quadrants (the sum of 9 10 degrees sectors): superior, nasal, inferior, and temporal. RESULTS: The mean intraocular pressure was 22.6 +/- 6.9 mm Hg preoperatively and 10.2 +/- 3.7 mm Hg postoperatively (P < 0.01). According to the analyses in every 10 degrees, the postoperative nerve fiber layer thickness was significantly greater than the preoperative nerve fiber layer thickness in the superotemporal region (10-50 degrees) and inferotemporal region (290-340 degrees) of the optic disc (P < 0.05). A stepwise multiple regression analysis showed that only the preoperative mean deviation in the Humphrey visual fields was a significant independent factor associated with changes in the nerve fiber layer thickness in the nasal and inferior quadrants. CONCLUSIONS: The thickness of the nerve fiber layer, as measured by scanning laser polarimetry, may increase after trabeculectomy, especially in the superotemporal and inferotemporal regions, and can be expected in cases in the early stage of glaucoma that have a better mean deviation. PMID- 11039739 TI - Motion-Evoked pattern visual evoked potentials in glaucoma. AB - PURPOSE: To evaluate motion-evoked brain potentials for glaucoma diagnosis. PATIENTS AND METHODS: Stripe patterns were presented in Maxwellian view under different stimulus conditions not combined in a factorial design. Spatial frequencies of 0.33 and 0.88 cycles/degree, with speeds of 10 and 5.9 degrees/second and contrasts of 0.04 and 0.93 were used. A 32 degrees whole field and a peripheral 32 degrees-27 degrees annular stimulus were used. Duration of motion was 200 milliseconds, and the interstimulus interval was 1,800 milliseconds. Recordings were obtained from Oz and P3. Thirty-four healthy patients, 12 glaucoma suspects, and 26 patients with open-angle glaucoma were tested. RESULTS: Normal response amplitudes decrease with age only under low contrast conditions, whereas response peak times increase under most conditions. Normal responses are much larger at P3 than at Oz, whereas in open-angle glaucoma, much less difference is seen. In these patients, the response amplitude at P3 is significantly reduced under all conditions, whereas a delay in peak time is less pronounced. A small but significant negative correlation (r = -0.44, P < 0.05) between response amplitude and mean perimetric defect was observed only with the annular stimulation. At a specificity of 90%, a sensitivity of approximately 76.7% for the low contrast and low spatial frequency condition was observed. CONCLUSIONS: Motion-evoked responses recorded at P3 are altered in open angle glaucoma and thus can be useful as an additional test in glaucoma diagnosis. PMID- 11039740 TI - Efficacy and safety of inferior 180 degrees goniosynechialysis followed by diode laser peripheral iridoplasty in the treatment of chronic angle-closure glaucoma. AB - PURPOSE: To report the efficacy and safety of inferior 180 degrees goniosynechialysis followed by diode laser peripheral iridoplasty in the treatment of chronic angle-closure glaucoma with total synechial angle closure. METHODS: Five patients with chronic angle-closure glaucoma and total synechial angle closure whose intraocular pressures were higher than 21 mm Hg while taking maximally tolerated medications underwent goniosynechialysis followed by diode laser peripheral iridoplasty to the inferior half of the angle. Intraoperative complications, postoperative visual acuity, intraocular pressures, and complications were evaluated. RESULTS: Five eyes of five patients received the operation and the mean follow-up was 7.6 months (range, 6-12 months). The mean preoperative intraocular pressure was 33.8 +/- 5.8 mm Hg. The mean postoperative intraocular pressure at most recent follow-up was 15.8 +/- 2.2 mm Hg. Postoperative complications included transient increase in intraocular pressure, hyphema, and cataract. The success rate (intraocular pressure less than 20 mm Hg with or without medication) was 80.0%. CONCLUSION: It appears that 180 degrees goniosynechialysis followed by diode laser peripheral iridoplasty is an effective and safe surgical procedure for treating chronic angle-closure glaucoma with total synechial angle closure. PMID- 11039741 TI - Conjunctival dysfunction and mitomycin C-induced hypotony. AB - PURPOSE: To determine the role of a physically intact conjunctiva in the development of chronic hypotony after mitomycin C-enhanced trabeculectomy. METHOD: Three patients with mitomycin C-related hypotonic maculopathy, but without a leak on Siedel test, had a thorough evaluation of the bleb area and an anterior segment fluorescein angiography. The bleb was excised and a pedicle flap, rotated from the temporal conjunctiva, was sutured to cover the defect superiorly. The scleral flap and its sutures were not disturbed. The excised bleb was subjected to light and electron microscopy. RESULTS: The Seidel test result was negative in all patients, but late phases of the anterior segment angiography showed a generalized seepage of aqueous from the bleb. After revision of the bleb, there was a gradual increase in the intraocular pressure, a reversal of the hypotonic maculopathy, and consequent improvement in visual acuity in all three patients, stable up to a minimum follow-up of 18 months. On histopathologic examination, the basement membrane was thickest under thin areas of the epithelium and thinnest below thicker epithelial layers. CONCLUSION: A dysfunctional conjunctival barrier, as evidenced by the "sweating" of the bleb and histopathologic alterations in the epithelial barrier, could be responsible for the hypotonic maculopathy in these patients. Excision of the conjunctiva alone and replacement by a pedicle conjunctival graft offers a safe and effective method of treating chronic hypotony after mitomycin C-augmented trabeculectomy in such patients. PMID- 11039743 TI - Neovascular glaucoma and ocular ischemic syndrome. PMID- 11039742 TI - Comparison of the clinical success rates and quality of life effects of brimonidine tartrate 0.2% and betaxolol 0.25% suspension in patients with open angle glaucoma and ocular hypertension. Brimonidine Outcomes Study Group II. AB - PURPOSE: To compare the clinical effectiveness and the impact on quality of life of twice-daily brimonidine 0.2% with those of twice-daily betaxolol 0.25% in patients with glaucoma or ocular hypertension. METHODS: A prospective, double masked, randomized, comparative, multicenter, 4-month "real-life" clinical trial involving 188 patients. Medications were instilled twice daily. Efficacy was determined through measurement of intraocular pressure; safety and tolerability were measured using reports of adverse events, a quality of life survey (Glaucoma Disability Index), heart rate, and blood pressure. Patients with an inadequate response in intraocular pressure after 1 month or those who experienced significant adverse events in the first month were switched to the alternative study arm and remained taking the alternative medication for a total of 4 months or left the study. The main outcome measure was clinical success, as determined by evaluation of the efficacy, safety, and tolerability of drug treatment, and was achieved when the investigator recommended that the patient continue the treatment after completion of the study. RESULTS: As initial therapy, clinical success was achieved in 74% of patients treated with brimonidine, as compared with 57% of patients treated with betaxolol (P = 0.027). The overall mean decrease in intraocular pressure from baseline was 5.9 mm Hg with brimonidine and 3.8 mm Hg with betaxolol. Both treatments were well tolerated, and there were no significant between-group differences in the incidence of adverse events or in the quality of life summary scores. CONCLUSIONS: Twice-daily brimonidine 0.2% and betaxolol 0.25% suspension were safe and effective as first-line therapy for glaucoma and ocular hypertension. In this study, brimonidine showed clinical effectiveness superior to that of betaxolol. PMID- 11039745 TI - Orthopaedic gene therapy. PMID- 11039744 TI - Religious fasting and intraocular pressure. AB - PURPOSE: To determine the effects of Islamic religious fasting on intraocular pressure. METHODS: The authors measured diurnal intraocular pressure values during religious fasting and 1 month later in 38 healthy male volunteers with a mean age of 22.4 +/- 2.7 years. Body weight and urine specific gravity were determined to assess the extent of dehydration caused by fasting. RESULTS: Although each patient had weight loss (0.1-1.4 kg) representing a slight dehydration caused by fasting, intraocular pressure values were not statistically different between fasting and nonfasting periods (P > 0.05). CONCLUSION: Fasting did not alter diurnal intraocular pressure values in healthy people. PMID- 11039746 TI - Studies on the chemical nature of the substance inducing transformation of pneumococcal types: Induction of transformation by a desoxyribonucleic acid fraction isolated from Pneumococcus type III. Oswald Theodore Avery (1877-1955) PMID- 11039747 TI - Genes and gene expression. AB - Gene therapy is the process whereby a therapeutic protein is synthesized from a DNA molecule (gene) that has been inserted into the cells. The goal is to produce the desired protein in the proper quantity in the proper location. Successful designing of vectors for gene therapy requires knowledge of gene structure and regulation. The gene is comprised of protein-coding sequences (called exons) that are interrupted by noncoding sequences (called introns). The expression of a gene in a tissue is regulated by proteins that bind to specific deoxyribonucleic acid sequences generally upstream from the first exon, in the deoxyribonucleic acid promoter domain. Understanding the mechanisms of gene regulation provides the deoxyribonucleic acid sequences necessary to direct expression of specific genes in the right tissue at the proper time in the desired amount. PMID- 11039748 TI - Vector systems for gene transfer to joints. AB - The prospects for the development of gene therapy treatments for certain orthopaedic diseases have been fueled by advances in the understanding of the molecular components of these disorders. These studies have identified molecules that could have therapeutic or reparative effects in certain settings. The ability to transfer and appropriately express the genes encoding these molecules is dependent on the availability of effective gene transfer vectors. Numerous vector systems have been used to transfer and express genes in joints with varied levels of success. The current review is designed to briefly outline the basics of the different gene transfer vector systems available for use by researchers in the orthopaedic fields. PMID- 11039749 TI - The adeno-associated virus vector for orthopaedic gene therapy. AB - During the last decade researchers working with recombinant adeno-associated virus have shown the use of this vector for efficient and long-term gene transfer in various tissues including lung, muscle, brain, spinal cord, retina, and liver. In 1999 the first results documenting the use of this vector in transducing joint cells were published. Additional advantages of recombinant adeno-associated virus for in vivo gene therapy are: (1) its ability to transduce nondividing cells; (2) site-specific integration into the host genome; (3) high viral titer (> 10(13)/mL); and (4) the vector is not cytotoxic and does not provoke a significant immune response. Most important, several groups have documented the ability to deliver sustained transgene expression in an immunocompetent host for more than 1 year, and that curative levels of gene product (factor IX), from one injection is sustained long-term in a large animal (hemophilia B dog). Comparable results have not been achieved with any other vector to date. As a result of this work the first Phase I clinical trials using recombinant adeno-associated virus are under way for cystic fibrosis. The history of the recombinant adeno associated virus vector and its future promise for orthopaedic gene therapies are described. The goal of the current review is to provide the reader with an understanding of the advantages and disadvantages of this vector for treatment of musculoskeletal diseases. Additional information concerning recombinant adeno associated virus can be obtained in more general reviews. PMID- 11039750 TI - Human genetic insights into skeletal development, growth, and homeostasis. AB - This current study describes how human genetic approaches are used to identify and understand the roles of genes and their protein products, during skeletal development, growth, and homeostasis. Searches for the genes responsible for brachydactyly, symphalangism, spondyloepiphyseal dysplasia, and camptodactyly arthropathy syndrome are presented to exemplify these approaches. The important role of the orthopaedic surgeon in this process is discussed. PMID- 11039751 TI - Gene therapy: clinical considerations. AB - Gene therapy can be defined as the introduction of nucleic acid into cells to ameliorate a disease process. To date there have been more than 313 trials with more than 2000 patients enrolled. The majority of these trials are Phase I or Phase II and have target diseases of either cancer or acquired immunodeficiency syndrome using retroviral and retroviral vectors. The choice of molecular target and the means of delivery have varied and chosen on the basis of the specific indication. Until recently the risks associated with treatment had been under appreciated. The first fatality associated with gene therapy occurred in September 1999 in which an adenoviral vector was used in the treatment of a patient with ornithine transcarbamylase deficiency. Subsequent to this report, other reports have surfaced suggesting that reporting of previous non-fatal reactions may have been minimized. Safety must be considered in relation to the disease process and to alternative treatments available. It may be easier to rationalize placing patients at risk who are facing a fatal disease process without effective alternative therapies. The ultimate goal of gene therapy will be the injection of a vector that has a specific target cell and that will be regulated by physiologic signals. Such a goal will require major improvements in the currently available delivery systems or the development of novel vectors. PMID- 11039752 TI - The ethics of the introduction of gene therapy into orthopaedic practice. AB - Gene therapy has the potential to transform musculoskeletal medicine. Orthopaedists have been ready to incorporate innovations in medicine and engineering into their surgical practice, frequently before having full information. There is no reason to doubt the rapid acceptance of gene therapy by the orthopaedic profession. Caution is needed in incorporating gene therapy into standard practice because of the lack of knowledge and risks that are greater than for previous innovations. PMID- 11039753 TI - Orthopedic gene therapy. Disease targets. PMID- 11039754 TI - Mesenchymal stem cells and gene therapy. AB - Multipotent human mesenchymal stem cells can be isolated from bone marrow and expanded more than 1-billion-fold in cell culture without the loss of their stem cell capacity. In addition, human mesenchymal stem cells can be transduced with genes for reporter molecules or secreted, circulating cytokines; these genes can be inserted into the genomes of mesenchymal stem cells without affecting their stem cell capacity. Thus, the stage is set for the use of mesenchymal stem cells as curative agents in genetic disorders involving skeletal tissues. PMID- 11039756 TI - Hematopoietic stem cells. AB - Hematopoietic stem cells are rare cells found in the bone marrow, peripheral blood, placenta, and elsewhere that sustain the hematopoietic system. The current review will focus on the evolving views of the biology of these cells. Because hematopoietic stem cells have been studied and transplanted for several decades, lessons learned from studying these cells may provide useful paradigms for the study of mesenchymal stem cells. PMID- 11039755 TI - Mesenchymal stem cells as vehicles for gene delivery. AB - Mesenchymal stem cells contribute to the regeneration of mesenchymal tissues such as bone, cartilage, muscle, ligament, tendon, adipose, and marrow stroma. Transduction of mesenchymal stem cells from species other than humans is required for the development of disease models in which mesenchymal stem cells-based gene delivery is evaluated. Attempts to transduce mesenchymal stem cells from some species with amphotropic retroviral vectors were unsuccessful, leading to comparative mesenchymal stem cells transductions with xenotropic and gibbon-ape leukemia virus envelope-pseudotyped retroviral vectors. Human, baboon, canine, and rat mesenchymal stem cells were transduced optimally with amphotropic vector supernatants. In contrast, sheep, goat, and pig mesenchymal stem cells showed highest transduction levels with xenotropic retroviral vector supernatant, and rabbit mesenchymal stem cells were transduced optimally with gibbon-ape-enveloped vectors. Using a myeloablative canine transplantation model and gene-marked canine mesenchymal stem cells, the biodistribution of infused and ex vivo expanded mesenchymal stem cells were examined. The majority of transduced canine mesenchymal stem cells were found in the bone marrow samples. The current study shows the use of mesenchymal stem cells as a delivery vehicle for gene transfer studies, and validates the feasibility of delivering mesenchymal stem cells to the marrow compartment for stromal regeneration after cancer-associated cytotoxic therapies. PMID- 11039757 TI - Orthopaedic gene therapy. Stem cells for gene delivery. PMID- 11039758 TI - Osteoinductive applications of regional gene therapy: ex vivo gene transfer. AB - Gene therapy represents the new frontier of medical science. Currently, there are no completely satisfactory treatment options for bone repair problems such as fracture nonunion, revision total joint arthroplasty, tumor resections, and fusions of the spine. Autogenous bone grafts, allograft implants, and prosthetic implants have been used to treat these problems. However, there are significant limitations associated with these methods including limited supply and limited osteogenic potential. Gene therapy, involving the manipulation of endogenous cells to generate specific proteins, offers a potential solution for these problems. By transferring genes into cells at a specific anatomic site, the osteoinductive properties of growth factors can be used at physiologic doses for a sustained period to facilitate a more significant healing response. Successful gene therapy involves four key steps: transduction, transcription, translation, and expression. To achieve gene transduction of a target cell, gene therapy models use vectors to enhance the entry and expression of exogenous deoxyribonucleic acid into the target cell's nucleus. The transduction of a gene can be performed via either an ex vivo or an in vivo approach. Although there are many potential target cells for gene therapy, the specific anatomic site, the quality of the bone, and the soft-tissue envelope, will influence the selection of the target cells for regional gene therapy. Gene therapy vectors delivered to a treatment site in osteoconductive carriers have yielded promising results. Several investigators have shown exciting results using ex vivo and in vivo regional gene therapy in animal models. Comparative studies and human clinical trials have not yet been performed but are necessary to identify the optimal genes and dosages for each specific application of regional gene therapy. In the future, the treatment options for bone loss problems will represent a clinical continuum based on the anatomic site, the condition of the target tissue bed, and the desired duration of protein production. PMID- 11039759 TI - In vivo nonviral delivery factors to enhance bone repair. AB - The purpose of the current study was to provide a review of a method for the nonviral delivery of genes into a skeletal defect for the purpose of promoting bone repair. To treat fractures at risk for delayed unions or nonunions, the delivery of plasmid deoxyribonucleic acid by a three-dimensional structural matrix is described. When these gene activated matrices are placed within a wound site, repair fibroblasts are observed migrating into the matrix where they encounter the plasmid deoxyribonucleic acid, take it up, and produce the protein that was defined by the plasmid. In varied animal models including rats, dogs, and sheep, the delivery of plasmids for parathyroid hormone or bone morphogenetic protein promoted bone formation and the healing of critical size defects. These studies show that the delivery of deoxyribonucleic acid to wound repair cells by three-dimensional matrix creates a persistent expression of factors that can promote bone formation. PMID- 11039760 TI - Potential role of direct adenoviral gene transfer in enhancing fracture repair. AB - Gene therapy has much to offer in the treatment of conditions in which it is necessary to increase the formation of bone. Nonunions, segmental defects, and aseptic loosening are examples of conditions where the local expression of genes that inhibit osteolysis and promote osteogenesis might be helpful. Studies in which one such possibility has been evaluated experimentally are described. These investigations used a surgically produced segmental defect in the femurs of New Zealand White rabbits as the model system. Adjacent muscle was fashioned around the defect to form a chamber into which adenoviral vectors were injected. High levels of transgene expression were found in the muscle surrounding the defect after injection of vectors carrying marker genes. Transgene expression also was seen in the cut ends of the bone and the scar tissue within the gap. No transgene expression was seen in the contralateral limb, spleen, or lung; transient, low levels of expression were found in the liver. Transgene expression declined with time, disappearing from all tissue but bone by Day 26; expression persisted in bone for at least 6 weeks. The control defects did not heal spontaneously. Injection of adenovirus carrying a human bone morphogenetic protein-2 complementary deoxyribonucleic acid led to healing of the segmental defect within 12 weeks, as judged by radiographic, histologic, and biomechanical criteria. Adenovirus carrying a human transforming growth factor-beta 1 complementary deoxyribonucleic acid showed signs of improved healing, but not to the extent seen with the bone morphogenetic protein-2 complementary deoxyribonucleic acid. This approach to therapy holds much promise as a novel means of promoting osteogenesis. PMID- 11039761 TI - Potential of gene therapy for treating osteogenesis imperfecta. AB - Osteogenesis imperfecta is a heterogeneous group of genetic disorders that affect connective tissue integrity, with bone fragility being the major clinical feature. Most forms of osteogenesis imperfecta are the result of mutations in the genes that encode the pro alpha1 and pro alpha2 polypeptide chains of Type I collagen. Because osteogenesis imperfecta is an incurable genetic disease, cell therapy and gene therapy are being investigated as potential treatments. Gene therapy for osteogenesis imperfecta however is a major challenge; because most of the mutations in osteogenesis imperfecta are dominant negative, supplying the normal gene without silencing the abnormal gene may not be beneficial. Null mutations in which an allele is not expressed or absent may be amenable to gene therapy or alternatively after silencing a mutant allele, a normal gene could be supplied. In addition, overexpression of the normal collagen gene in cells expressing mutant collagen polypeptide chains potentially could lead to synthesis of a sufficient percentage of normal molecules to normalize clinical status. The authors currently are examining the possibility of developing gene therapy for treating a mouse model of human osteogenesis imperfecta (oim) using bone marrow stromal cells as vehicles for delivering normal collagen genes to bone. In the current study, the potential of gene therapy for treating osteogenesis imperfecta is discussed in the context of the complexity of the mutations in Type I collagen genes that lead to different osteogenesis imperfecta phenotypes. PMID- 11039762 TI - Pluripotential mesenchymal cells repopulate bone marrow and retain osteogenic properties. AB - Precursor cells, isolated from bone marrow, can develop into various cell types and may contribute to skeletal growth, remodeling, and repair. The D1 cell line was cloned from a multipotent mouse bone marrow stromal precursor and has osteogenic, chondrogenic, and adipogenic properties. The osteogenic phenotype of these precursor cells is relevant to the process of fracture healing and osteointegration of prosthetic implants. The D1 cells were labeled genetically using a replication incompetent retroviral vector encoding beta-galactosidase, an enzyme which is used as a marker. Labeled cells are readily identifiable by staining with 5-bromo-4-chloro-3-indoyl-beta-D-galactoside and by flow cytometry, and retain the desired osteogenic characteristics in vivo as shown by von Kossa staining, alkaline phosphatase assay, an increase in cyclic adenosine monophosphate in response to parathyroid hormone, osteocalcin messenger ribonucleic acid production, and bone formation in diffusion chambers. In addition, the cells cloned from marrow stroma repopulate the marrow of host mice, persist for several weeks, and retain their osteogenic potential ex vivo. The data suggest that such cells may be used to replenish the number of osteoprogenitors in marrow, which appear to decrease with age, thereby leading to recovery from bone loss and improved bone growth and repair. Labeling these cells creates a model in which to study the potential of such cells to participate in fracture repair, ingrowth around prosthetic implants, treatment of osteoporosis, and to explore the possibility of gene delivery to correct mutations or defects in metabolism that are responsible for certain skeletal abnormalities. PMID- 11039763 TI - Marrow transplantation and targeted gene therapy to the skeleton. AB - Treatment of genetic or degenerative diseases severely affecting the entire skeleton may necessitate gene therapy involving transplantation of multipotential marrow cells. The ability of in vitro expanded adherent marrow cells enriched in pluripotent mesenchymal cell populations to remain competent to engraft, repopulate host tissues, and differentiate into bone and cartilage is advantageous for correction of skeletal-related diseases. However, to achieve phenotypic specificity and therapeutic or physiologic levels of proteins may require cell type specific expression of the gene. Tissue-specific promoter controlled transgenes provide an efficacious approach to deliver therapeutic gene expression to repopulating chondrocytes and osteoblasts for treatment of cartilage and bone disorders or tumor metastasis to the skeleton. The bone specific expression of a reporter gene controlled by the osteoblast-specific osteocalcin promoter after transplantation of a mixed population of marrow cells is shown. Tissue-restricted gene therapy potentially can be refined by use of a unique peptide targeting signal that directs the hematopoietic, chondrogenic, and osteogenic core binding factor/acute myelogenous leukemia transcription factors to subnuclear sites that support gene expression. PMID- 11039764 TI - Orthopaedic gene therapy. Fracture healing and other nongenetic problems of bone. PMID- 11039765 TI - Gene therapy for genetic diseases of bone. PMID- 11039766 TI - Chondrogenitor cells and gene therapy. AB - The development of isolation and culture techniques for mesenchymal progenitor cells from various tissues has promoted interest in the use of these cells for repair and regeneration of musculoskeletal tissues. The chondrogenic differentiation of these pluripotential cells seems to be mediated by numerous cytokines most of which belong to the transforming growth factor-beta superfamily. Strategies to repair articular cartilage have focused on delivery of these cytokines or progenitor cells to the area of damage. More recently, with the development of gene transfer techniques, these cells have become the target of in vivo gene therapy, which involves direct injection of viral and nonviral vectors carrying transgenes. Furthermore, they are viewed as potential carriers of the transgenes for ex vivo gene therapy, in which the gene transfer is done in vitro with culture-expanded cells that then are implanted or injected. In vitro data suggest that the chondrogenic potential of these cells is maintained with virally mediated ex vivo gene transfer. By transducing these cells with chondroinductive factors, the bioactive factors and the target cells are delivered to the repair site. PMID- 11039767 TI - Cartilage and bone regeneration using gene-enhanced tissue engineering. AB - Joint cartilage injury remains a major problem in orthopaedics with more than 500,000 cartilage repair procedures performed yearly in the United States at a cost of hundreds of millions of dollars. No consistently reliable means to regenerate joint cartilage currently exists. The technologies of gene therapy and tissue engineering were combined using a retroviral vector to stably introduce the human bone morphogenic protein-7 complementary deoxyribonucleic acid into periosteal-derived rabbit mesenchymal stem cells. Bone morphogenic protein-7 secreting gene modified cells subsequently were expanded in monolayer culture, seeded onto polyglycolic acid grafts, implanted into a rabbit knee osteochondral defect model, and evaluated for bone and cartilage repair after 4, 8, and 12 weeks. The grafts containing bone morphogenic protein-7 gene modified cells consistently showed complete or near complete bone and articular cartilage regeneration at 8 and 12 weeks whereas the grafts from the control groups had poor repair as judged by macroscopic, histologic, and immunohistologic criteria. This is the first report of articular cartilage regeneration using a combined gene therapy and tissue engineering approach. PMID- 11039768 TI - Type II interleukin-1beta receptor: a candidate for gene therapy in human arthritis. AB - Interleukin-1 plays a pivotal role in the pathophysiologic process of arthritis. It is released in an autocrine fashion in joints affected with arthritis. Human cartilage affected with arthritis (but not normal cartilage) showed upregulation of interleukin-1beta messenger ribonucleic acid and protein in ex vivo conditions. Type II interleukin-1 receptor, interleukin-1 receptor antagonist, and Type I soluble interleukin-1 receptor has potent interleukin-1 neutralizing activity. In view of these observations, the role of type II interleukin-1 receptor in chondrocyte function was examined. Human interleukin-1beta (5-10 ng/mL) induced nitric oxide, prostaglandin E2 and matrix metalloprotease production in bovine or human chondrocytes, which could be inhibited by 500 pg/mL of Type II interleukin-1 receptor. Interleukin-1 inhibited proteoglycan synthesis that could be reversed by Type II soluble interleukin-1 receptor. Similarly, 1 ng/mL human interleukin-1 induced (or spontaneously produced) nitric oxide and prostaglandin E2 production in human osteoarthritis-affected cartilage could be inhibited by 50% or greater with 100 pg/mL Type II interleukin-1 receptor in ex vivo conditions. Type II interleukin-1 receptor transfected chondrocytes were immune to the insults of exogenous interleukin-1. These experiments showed that endogenous or exogenous Type II interleukin-1 receptor can attenuate the effects of interleukin-1 with respect to induction of inflammatory mediators, matrix metalloprotease activity and, proteoglycan synthesis in human chondrocytes and cartilage. PMID- 11039769 TI - Nonviral in vivo gene therapy for tissue engineering of articular cartilage and tendon repair. AB - Heretofore, nonviral methods have been used primarily for in vitro transfection of cultured cell lines. These methods were substantially less efficient when compared with the use of viruses, particularly when used in vivo. Herein a three step, highly efficient method of nonviral gene delivery is presented. Using this method, genes have been delivered successfully into tissues of orthopaedic importance with high-efficiency by nonviral means. Transforming growth factor beta 1, parathyroid hormone related protein, and a marker gene were transfected into primary perichondrium and cartilage cells with efficiencies in excess of 70%. They overexpressed their cognate gene products showing efficacy of expression in a rabbit model of osteochondral defect repair. Using the same method, a marker gene was delivered into a canine model for intrasynovial flexor tendon injury and repair. This was achieved by direct gene delivery during surgery. An estimated 5 additional minutes were required during surgery to complete the transfection steps. High efficiency gene delivery was achieved in the flexor tendons, tendon sheaths, tendon pulleys, surrounding tissues, and skin. The efficiency of transfection approached 100% in the exposed superficial tissue layers and transfected cells were found several layers below the exposed tissue surfaces. The data show the potential of direct nonviral gene therapy in orthopaedics for ex vivo and in vivo applications. PMID- 11039771 TI - Using gene therapy to protect and restore cartilage. AB - Numerous gene products have the potential to help protect cartilage from degradation and to repair cartilage that has become damaged as a result of disease or injury. The genes that encode these products thus may serve as chondroprotective and chondroregenerative medicines. To bring these agents into clinical use, it is necessary to screen candidate genes for efficacy under in vitro and in vivo conditions, to determine the best cells to target, and to develop appropriate gene transfer technologies. As discussed in the current review, progress has been made in each of these areas. Various viral and nonviral vectors are able to deliver genes to synoviocytes, articular chondrocytes, and mesenchymal stem cells. There also is evidence to suggest that ex vivo and in vivo approaches can be used for gene transfer to articular cartilage, synovium, and meniscus. Moreover various cytokine antagonists and growth factors have been shown to protect cartilage and stimulate chondrogenesis. In vivo methods and strategies that target synovium may be useful in a chondroprotective mode but because they do not increase the number of chondrogenic cells within lesions, they may be ill-equipped to repair large defects. Ex vivo methods however, provide cells and genes. It also is important to distinguish the treatment of isolated lesions occurring as a result of injury from the treatment of lesions resulting from underlying disease processes. Additional development of these approaches should result in clinically useful genetic methods for the protection and regeneration of cartilagenous tissues. PMID- 11039770 TI - Insulinlike growth factor-I gene therapy applications for cartilage repair. AB - Cartilage function after resurfacing with cell-based transplantation procedures or during the early stages of arthritic disease may be bolstered by the addition of growth factor genes to the transplanted tissue. Insulinlike growth factor-I maintains chondrocyte metabolism in normal cartilage homeostasis and has been shown to improve cartilage healing in vivo. Given the relatively short half-life of insulinlike growth factor-I in biologic systems, however, maintenance of effective concentrations of this peptide has necessitated either very high initial doses or repeated treatment. Delivery of the insulinlike growth factor-I gene, using a deleted adenovirus vector, specifically targeting graftable articular chondrocytes, bone marrow-derived chondroprogenitor cells, or synovial lining cells, may provide more durable insulinlike growth factor-I fluxes to articular tissues. Cultured equine articular chondrocytes, mesenchymal stem cells, synovial explants, and synovial intimal cells were readily transfected with an E1-deleted adenoviral vector containing equine insulinlike growth factor I coding sequence. Optimal viral concentrations for effective transduction were 100 multiplicities of infection in synoviocytes, 500 multiplicities of infection in chondrocytes, and 1000 multiplicities of infection in mesenchymal stem cells. Production of insulinlike growth factor-I ligand varied from 65 ng/mL to 246 ng/mL in medium from chondrocytes and synovial explants, respectively. For chondrocytes, these concentrations were sufficient to produce significant stimulation of cartilage matrix gene expression and subsequent proteoglycan production. Moreover, cells in infected cultures maintained a chondrocytic phenotype and continued to express elevated insulinlike growth factor-I levels during 28 days of monolayer culture. Minimal synthetic activity, other than insulinlike growth factor-I ligand synthesis, was evident in synovial cultures. These experiments suggest several avenues for insulinlike growth factor-I supplementation of articular cartilage, including preimplantation adenoviral insulinlike growth factor gene transfer to chondrocytes or chondroprogenitor cells, and direct injection of adenoviral-insulinlike growth factor to transfect the synovial structures in situ. PMID- 11039772 TI - Gene therapy for chondrocytes. PMID- 11039773 TI - Orthopaedic gene therapy. Chondroprogenitors. PMID- 11039774 TI - Gene therapy for spine fusion. AB - Spine fusion is a commonly performed yet often unsuccessful surgical procedure. As many as 40% of patients undergoing spine fusion may have a nonunion or failure to form a continuous bone bridge. This clinical challenge has focused much of the attention of osteoinductive bone growth factors toward spine applications. Clinical pilot and pivotal trials will show the feasibility of recombinant and purified bone growth factors to promote spine fusion in humans. Despite this, strategies of gene therapy for spine fusion and other bone healing applications are being pursued. This article reviews the state of the art of local gene therapy and highlights specific issues that must be addressed when pursuing a gene therapy program. Perhaps the most critical step in gene therapy for bone formation is choosing an appropriate osteoinductive gene. Such choices may be limited by differences in efficacy of the chosen gene and availability because of proprietary constraints. The choice of delivery vector is crucial and depends on the potency of the gene and the specific application intended. Establishing the effective dose, transduction time, and gene transfer method are important decisions. The choice of carrier material to form the scaffold for the new bone formation is paramount to successful bone formation. Finally, a strategy for in vitro and in vivo testing must be developed to maximize the chances of success in human trials. PMID- 11039775 TI - Potential applications of gene therapy to the treatment of intervertebral disc disorders. AB - Gene therapy involves the transfer of genes to cells such that the recipient cells express these genes and thereby synthesize the ribonucleic acid and protein that they encode. Recent investigations suggest that gene therapy may have potential applications in the treatment of intervertebral disc disorders, particularly those associated with disc degeneration. The successful in vivo transfer of therapeutic genes to target cells within the intervertebral disc in clinically relevant animal models is one example of the rapid progress that is being made. The purpose of the current review is to address several important technical issues, including choice of vectors and gene delivery strategy and the characteristics of the target tissues, which are relevant to future clinical applications of gene therapy for the treatment of intervertebral disc disorders. It already is apparent from the growing literature that gene therapy has the potential of becoming a valuable clinical treatment mode for intervertebral disc disorders in the twenty-first century. PMID- 11039776 TI - Complexity of determining cause and effect in vivo after antisense gene therapy. AB - Injuries to joint tissues are major clinical problems occurring with significant frequency and resulting in the formation of scar tissue or in some tissues with no healing at all. Such scar tissue has compromised biomechanical integrity, which leads to impaired function, increased risk of reinjury, induction of remodeling in other joint tissues and increases the risk of diseases such as ostheoarthritis. Development of new therapies, such as gene therapy, to enhance repair could have a significant impact on quality of life for patients. The well characterized rabbit medial collateral ligament injury model was used to transiently modulate the expression of specific molecules during early stages of healing. The small matrix proteoglycan decorin, known to influence matrix assembly and to bind and growth factors, was targeted in vivo using decorin specific antisense oligodeoxynucleotides and Hemagglutinating Virus of Japan Liposome method. After 4 weeks of healing, scar tissue was assessed after antisense exposure by reverse transcription polymerase chain reaction, Western Blot analysis, light and transmission electron microscopy, and biomechanically for low and high load behavior. Ligament scar messenger ribonucleic acid and protein levels for decorin decreased and collagen fibril diameter size increased after antisense treatment. Creep and stress at failure improved after antisense treatment indicating a functional improvement in the scar tissue. However, messenger ribonucleic acid levels for multiple genes were affected by the decorin specific antisense treatment and therefore all of the observed improvements in the scar tissue cannot be directly ascribed to depressing decorin levels. PMID- 11039777 TI - In vivo gene transfer into tendon by recombinant adenovirus. AB - Recombinant adenovirus mediated Escherichia coli lacZ gene transfer into chicken tendon and tendon sheath has been reported in the current study. The constructed recombinant virus carrying lacZ gene was injected between tendon and tendon sheath to conduct in vivo gene transfer. During the course of the study, each tendon received a 10 uL injection containing 10(5) plaque forming units of recombinant adenovirus with beta-galactosidase gene. The samples were harvested at 3 days, 30 days, and 75 days after injection. For the virus dose-transduction rate study, five different doses were injected to groups of chicken tendons. LacZ gene transfer was detected for its coding product beta-galactosidase by staining with X-gal solution. Results showed that the tendon and tendon sheath received the gene transfer with blue staining. The transferred lacZ gene remained stable for 75 days in the tendon and tendon sheath. A virus dose-dependent pattern of transduction rate was observed in the gene transferred tendons. The area of tendon transduction was approximately 2% for 3 x 10(6) plaque forming units recombinant adenovirus with beta-galactosidase versus 40% for 6 x 10(7) plaque forming units recombinant adenovirus with beta-galactosidase gene. The data suggested that functional exogene could be transferred into the tendon and tendon sheath by the same strategy to improve healing and avoid adhesion. PMID- 11039778 TI - Orthopaedic gene therapy. Spine. PMID- 11039779 TI - Orthopaedic gene therapy. Ligament and tendon. PMID- 11039780 TI - Gene therapy for osteoarthritis: new perspectives for the twenty-first century. AB - Morphologic changes observed in osteoarthritis include cartilage erosion and a variable degree of synovial inflammation. Proinflammatory cytokines such as interleukin-1 beta, locally produced by the inflamed synovium, also likely contribute to these alterations. Despite an extensive armamentarium and numerous surgical options, osteoarthritis remains incurable, and an improved approach in the treatment of this disease is imperative. Drug delivery is a major weakness of existing antiarthritic therapies. Local delivery of antiinflammatory cytokines or the in vivo induction of their expression using gene transfer may provide a novel approach for the treatment of osteoarthritis. Evidence of the efficacy of gene therapy in osteoarthritis remains very scarce. To the authors' knowledge, there is no clinical research protocol en route for the treatment of osteoarthritis using gene therapy. The authors present the only two studies that have proved successful in treating animal models of osteoarthritis using gene therapy, and propose an overview of several strategies for the development of gene therapy in osteoarthritis treatment in the future. PMID- 11039781 TI - Evaluation of gene therapy as a treatment for equine traumatic arthritis and osteoarthritis. AB - Joint disease in horses and in humans is a significant social and economic problem and continued research and improvements in therapeutics are needed. Because horses have naturally occurring osteoarthritis that is similar to that of humans, the horse was chosen as a species to investigate gene transfer as a potential therapeutic modality for the treatment of osteoarthritis. Using an established model of equine osteoarthritis, the therapeutic effects resulting from overexpression the equine interleukin-1 receptor antagonist gene sequence through adenoviral mediated gene transfer was investigated. The results of the current study showed intraarticular expression of interleukin-1 receptor antagonist to have favorable effects such as an approximately 28 day upregulation of interleukin-1 receptor antagonist protein expression, significant improvement in clinical parameters of pain and disease activity, and beneficial effects in histologic parameters measured from synovial membrane and articular cartilage when compared with nontransduced joints. Based on the significant improvements seen in this work gene transfer of interleukin-1 receptor antagonist is a practical treatment modality for the equine patient and also offers future promise for human patients. PMID- 11039782 TI - Gene therapy approaches for treating rheumatoid arthritis. AB - Current gene therapy approaches for treating rheumatoid arthritis have made use of gene transfer technology as an improved delivery system for emerging proteins and other biologicals whose activities may have therapeutic value. Preclinical research has focused on two primary directions, evaluation of methods of gene delivery and identification of gene products with antiarthritic potential. Although there are reports involving systemic gene delivery, the bulk of effort has focused on local, intraarticular administration using ex vivo and in vivo methods. Viral-based vectors, including adenovirus, adeno-associated virus and herpes simplex virus have the greatest efficiency of gene delivery after intraarticular injection and are capable of generating relevant levels of gene products in several animal models of disease. However, there are limitations to existing generations of these systems that currently preclude their clinical application. Those gene products found to be efficacious in animal models of rheumatoid arthritis include proteins that specifically block the activity of the primary inflammatory cytokines, and include interleukin-1 receptor antagonist and soluble receptors for tumor necrosis factor and interleukin-1. Delivery and expression of genes encoding certain cytokines such as interleukins -4, -10, and 13 and viral interleukin-10, that block synthesis of inflammatory mediators and downregulate aspects of cellular and humoral immune pathways have been found beneficial. Although significant progress has been made, leading to Phase I clinical trials, there remain several hurdles to the routine practice of gene therapy for treatment of rheumatoid arthritis. PMID- 11039784 TI - Gene therapy for rheumatoid arthritis. PMID- 11039783 TI - Clinical trials in the gene therapy of arthritis. AB - Gene therapy clinical trials raise important safety issues that complicate their design and require extensive preclinical testing. Human protocols for the treatment of arthritis and most other orthopaedic and rheumatologic indications are complicated additionally by the perception that they are largely acquired, nonlethal conditions. Taking these considerations into account, the first such human study used the local, ex vivo delivery of a gene whose product, the interleukin-1 receptor antagonist, has an outstanding safety profile. This gene was delivered to the metacarpophalangeal joints of postmenopausal women 1 week before these joints were removed during total joint replacement surgery. In addition to providing an additional safety cushion, the surgical removal of the genetically modified joints made available large amounts of tissue to examine for evidence of successful gene transfer and gene expression. This Phase I safety study was approved at the local and federal levels, and its funding was contingent on the establishment of an external monitoring board. This trial now has been completed and a Phase II, efficacy study is being planned. A similar study has begun in Dusseldorf, Germany and results from the first two patients are similar to the results of the American patients. Permission has been given for two additional human trials, one in the United States and one in the Netherlands, in which a gene encoding herpes thymidine kinase will be transferred to the joints of patients with rheumatoid arthritis who then will be administered gancyclovir. This procedure aims to treat the disease by producing a genetic synovectomy. Additional development of human gene therapies for arthritis and other orthopaedic and rheumatic conditions will be aided by the successful completion of these studies. PMID- 11039785 TI - Orthopaedic gene therapy. Osteoarthritis. PMID- 11039786 TI - Total hip arthroplasty using a ceramic prosthesis. Pierre Boutin (1924-1989). PMID- 11039787 TI - Alternate bearing surfaces in total joint arthroplasty: biologic considerations. AB - The problem of periprosthetic osteolysis is currently the major limiting factor in joint arthroplasty longevity. Because this process has been shown to be primarily a biologic response to wear particles, corrosion products, or both, efforts to reduce particle generation are being undertaken. These efforts include the development of modified polyethylene and alternative articulating surfaces. These alternate bearing surfaces currently include ceramic-on-polyethylene, ceramic-on-ceramic, and metal-on-metal. Although these alternate bearings diminish or eliminate the generation of polyethylene particles, ceramic and metal particles are produced. The purpose of the current review is to discuss the literature that addresses the biologic response to these particles, locally and systemically. PMID- 11039788 TI - Ceramic femoral head retrieval data. AB - In the 1970s it was first realized that the properties of alumina ceramics could be exploited to provide better implants for orthopaedic applications. Applications depend on the fact that ceramics provided wear characteristics suitable for bearing surfaces in total hip replacement. Resultant orthopaedic use had more than 20 years' clinical success. To date more than 2.5 million alumina femoral heads have been implanted. Published reports of fracture rates of the alumina heads range between 0% for ceramics manufactured after 1990 and 13.4% for ceramics manufactured before 1990. These high fracture rates were caused by materials manufactured by companies that are not on the market today. These old aluminas had a low density, had a very coarse microstructure, and were not in compliance with specifications that are valid today. Materials scientists have substantially improved the mechanical strength of alumina. There are three generations of medical grade aluminas. The latest generation is an alumina that is hot isostatic pressed, laser marked, and proof tested. This material has been on the market since 1994. The fracture rates of the most commonly used ceramics have been analyzed by various groups, and are based on .5 million femoral heads to 1.5 million femoral heads. The fracture rate of ceramic Biolox femoral heads are 0.026 % for first generation alumina, 0.014% for second generation alumina, and 0.004% for femoral heads manufactured after 1994. Analyzing the clinical experience of more than 20 years, it can be concluded that all the improvements have produced reliable ceramic femoral heads. PMID- 11039789 TI - Ceramic component failure and the role of proof testing. AB - Proof testing is an overload test used for high demand materials that must have 100% of the components tested to sort out those components with possible internal flaws. The relationship of high proof loads applied short-term, low service load, and service time of proof tested components is shown. Proof testing is now technically and clinically validated. Ceramic femoral heads that have been proof tested have been used clinically for 4 years and a significant reduction of the failure rate from 0.015% to 0.004% has resulted. However, component service life is critical and has to be protected in unison with the prosthesis manufacturer and the hospital. Because ceramics are rigid, joining of ceramic components with metals requires special attention. Specifications of the tapers and/or connection surfaces have to be appropriate to the needs of the ceramic and the manufacturer of ceramic components and the prosthesis manufacturer must adhere to strict specifications and inspection techniques. In the surgical area, training in the proper use of ceramics is a key factor to the successful performance of ceramic components. PMID- 11039790 TI - Clinical and hip simulator comparisons of ceramic-on-polyethylene and metal-on polyethylene wear. AB - The benefit of reduced polyethylene wear with ceramic in hip replacements does not seem to have been universally appreciated. In this current study, wear predictions from laboratory and clinical studies were compared for ceramic-on polyethylene and cobalt chrome-on-polyethylene combinations. Many laboratory studies included water-based lubrication and linear-tracking mechanisms. Now it is appreciated that these were inappropriate methods, because of a propensity for very low or virtually no polyethylene wear against ceramics in water. Thus, water based studies predicting a 20- to 80-fold advantage for ceramic-on-polyethylene compared with metal-on-polyethylene clearly were in error. However, serum-based simulator studies with high protein-concentrations generally have shown greater wear with alumina-on-polyethylene than with metal-on-polyethylene. Controversy still remains over the use of such nonphysiologic protein levels. The simulator studies were just beginning to explore the role of serum protein concentrations and the influence on the various wear models. Polyethylene wear with zirconia systems was particularly affected by serum protein concentrations. In one simulator study, use of proteins in the physiologic range resulted in the alumina on-polyethylene wear rate decreasing to approximately 50% of that of metal-on polyethylene. In the literature, many hip design and polyethylene variations were reported which confounded the wear analysis. Overall, the clinical data supported the superior performance of ceramic-on-polyethylene systems by a factor of 1.5- to fourfold. However, the amount of supporting data was not large. This summary of laboratory and clinical data indicated that ceramic-on-polyethylene hip replacement systems offered on average a 50% wear reduction from metal-on polyethylene systems. PMID- 11039791 TI - Modern ceramic-on-ceramic total hip systems in the United States: early results. AB - In 1997, two manufacturers began Food and Drug Administration approved investigations of a ceramic-on-ceramic (alumina) articulation total hip replacement in the United States. Osteonics (Allendale, NJ) and Wright Medical Technology (Arlington, TN) enrolled more than 500 and 300 patients, respectively, when their studies ended in the middle part of 1998. The author presents detailed early results of the series by Wright Medical Technology. Three hundred thirty three patients were enrolled in 11 centers around the country in a prospective series. All patients received the Transcend ceramic-on-ceramic articulation and have a minimum of 18 months followup with a range of 18 to 36 months. Harris hip scores increased on average from 44 to 97 points. Four patients underwent revision surgery; one for deep infection, one for early migration of the cup, one for dislocation, and one for liner malplacement. Overall, there were 42 complications. Seven were systemic and 35 were related to the total hip replacement. Four of the 35 complications were ceramic related and included three chipped liners and one eccentric seating of the cup liner. To date, no patient underwent revision surgery for aseptic loosening. Seven technical guidelines are suggested to enhance the quality of the intraoperative and postoperative results: a conservative femoral neck cut; horizontal cup placement (< 45 degrees); increased anteverted cup placement (> 20 degrees); use of trial liners; impaction of ceramic pieces; hand placement of the liner; and removal of osteophytes and/or part of the anterior wall of the acetabulum to avoid impingement. Alternate bearing articulations, particularly ceramics, have important technical aspects to be considered at the time of implantation to minimize intraoperative and postoperative complications. To date, there have been no postoperative fractures of the ceramic pieces in any of the completed or ongoing ceramic-on-ceramic investigations by all involved manufacturers. These preliminary results are satisfactory at this time. One can look to the future with cautious optimism. PMID- 11039792 TI - Evolution of alumina-on-alumina implants: a review. AB - The use of alumina-on-alumina sliding surfaces for total hip replacement is becoming increasingly popular. The author has reviewed the 30-year history of this material. Technical aspects such as the quality of the material, quality of the design, and the risk of fracture are presented. The clinical results observed by the author are summarized with additional references to the results of other surgeons. The overall conclusion is that this material is very safe if all the quality requirements are met. The extremely low generation of wear debris and the excellent biologic tolerance of the material impart a long lifetime to the implant in young and active patients. PMID- 11039793 TI - Ceramic hip prostheses in young patients: a retrospective study of 74 patients. AB - Zones of osteolysis occur in mobilized and in well-fixed prostheses with the classic ultrahigh molecular weight polyethylene-metal couple. However, since 1981 with the use of the ceramic couple in cemented hip prostheses in young patients, osteolysis has not been observed. The prostheses in patients with a longer followup do not show signs of wear in the bearing or signs of bone lysis in the well-fixed prostheses. Cementless prostheses with the same type of ceramic bearing have been used since 1990 in patients with more than 30 years of life expectancy, in an effort to eliminate loosening at the cement-bone interface. The cementless prostheses performed well with spot welds visible 3 months after the operation, which still are present at 7 years of followup. Lysis has not been observed behind the cup or around the stem. In the current series, head ruptures and accidents in the stem and neck junction produced by the modularity of the prostheses did not occur. The authors focus on the positive results shown by ceramic bearings and describe the results of 31 cemented, five hybrid, and 58 uncemented prostheses with ceramic bearings in patients younger than 50 years of age. PMID- 11039794 TI - Alumina-on-alumina total hip prostheses in patients 40 years of age or younger. AB - To avoid the consequences of polyethylene wear in a high-risk population, 128 alumina-on-alumina total hip arthroplasties have been done in 104 consecutive patients. The maximum age of patients was 40 years. The main preoperative diagnoses were osteonecrosis and sequellae of congenital hip dislocation (71% of the hips). The same titanium alloy cemented stem was implanted in all of the hips. Four types of alumina acetabular component fixations were used: a cemented plain alumina socket (41 hips), a screw-in ring with an alumina insert (22 hips), a press-fit plain alumina socket (32 hips), and a press-fit titanium metal back with an alumina insert (33 hips). Eight patients (11 hips) died during the followup period. Sixteen revisions have been documented, 12 for acetabular aseptic loosening, three for bipolar loosening (two of which were septic), and one for unexplained pain. Eighty-eight hips in 74 patients have been followed up radiologically for 2 to 22 years. Wear was unmeasurable. Four additional sockets showed definite migration. The respective survival rates after 7 years were 94.1% for the cemented cup, 88.8% for the screw-in ring, 95.1% for cementless press-fit plain alumina socket and 94.3% for the metal-back press-fit component. The 10 year survival rate was 90.4% for the cemented socket and 88.8% for the screw-in ring. The 15-year survival rate was 78.9% for the cemented socket. Grafting was the only prognostic factor, with a survival rate of 62.6% after 10 years for the hips with a bone graft and of 90.1% for hips without a graft. The alumina-on alumina bearing surfaces seem to be a valuable alternative to the standard metal on-polyethylene system for young patients. However, an improvement in socket fixation is required to lengthen the life span of the prosthesis to match the life expectancy of this demanding population. PMID- 11039795 TI - Clinical experience with ceramics in total hip replacement. AB - As part of a search for better articulation in total hip prostheses, the decrease in the thickness of the socket in different total hip prostheses was measured in vivo. The wear rates of (1) RCH 1000 (molecular weight, 10(6)) socket gamma irradiated with 100 Mrad articulating with a crude COP (stainless steel containing 20% cobalt and 0.01% phosphorous) metal femoral head; (2) RCH 1000 socket nonirradiated articulating with a crude COP femoral head; (3) RCH 1000 socket irradiated with 100 Mrad articulating with an alumina femoral head; (4) ultra high molecular weight polyethylene (molecular weight, 5-6 x 10(6)) socket articulating with an alumina femoral head; and (5) ultrahigh molecular weight polyethylene socket articulating with a stainless steel femoral head (T-28) were 0.06, 0.30, 0.06, 0.1 and 0.25 mm/year, respectively, in the authors' clinical cases. Alumina femoral heads were effective in decreasing wear of the polyethylene socket. However, the wear rates of gamma-irradiated sockets articulating with alumina and with metal femoral heads wear very low and were not different from each other. Regarding the relationship between wear rate and the thickness of the ultra high molecular weight polyethylene socket articulating with a 28 mm alumina femoral head, on radiographs, average wear rates of socket thicknesses of 7, 8, 9, 10 and 11 mm were 0.14, 0.15, 0.12, 0.06, and 0.08 mm/year, respectively. On measuring retrieved prostheses, average wear rates of 7, 8, 9 and 11 mm thickness sockets were 0.2, 0.19, 0.14, and 0.1 mm/year, respectively. The wear of sockets has been proven to be minimal in alumina femoral heads articulating with ultrahigh molecular weight polyethylene sockets thicker than 10 mm. PMID- 11039796 TI - Alumina ceramic bearings for hip endoprostheses: the Austrian experiences. AB - The current authors review clinical and retrieval experiences with hemispheric monolithic alumina ceramic sockets (Group 1), implanted between 1976 and 1979, and similar modular titanium sockets with alumina ceramic inlays (Group 2), implanted between 1990 and 1995. Both cementless sockets articulated with alumina ceramic femoral ball heads for total hip joint replacements. Clinical followup of patients with hemispheric monolithic alumina ceramic sockets (Group 1, 138 sockets) resulted in a total failure rate of 19.6% after 5 to 20 years. Radiologic analysis of eight stable sockets showed migration of 0.2 mm to 2.89 mm, but in four sockets at risk for late aseptic failure after an average followup of 12.5 years as much as 13.4 mm of migration was seen. Histologic evaluation revealed pseudosynovial membranes as thick as 1 mm with fine birefringent wear particles within mononuclear macrophages around two stable retrieved sockets. The membranes around four loose sockets were 6 to 10 mm thick and also heavily loaded with larger alumina wear particles. After 7 years followup clinical analysis of patients with modular titanium sockets with alumina ceramic inlays (Group 2, 30 sockets) resulted in four revisions, compared with one revision of 50 identical sockets (control group) with polyethylene instead of alumina ceramic inlays. Wear particle analyses in scanning electron microscopy showed significantly more particles (x 10(9) +/- standard deviation/g dry tissue) from the control group (4.26+/-6.38), compared with alumina ceramic bearings of Group 1 (0.70+/-0.79), and of Group 2 (1.62+/-2.13). The alumina particle sizes ranged between 0.13 and 78.38 microm. The mean annual linear wear of 38.8 microm was calculated for the bearings in Group 1, and of 26.94 microm for bearings in Group 2. These results support the good tribologic and biologic performance of alumina ceramic bearings for total hip arthroplasty. PMID- 11039797 TI - Zirconia as a sliding material: histologic, laboratory, and clinical data. AB - Zirconia ceramics have been introduced in orthopaedic surgery as prosthetic femoral heads to solve the critical issue of femoral head fractures sometimes observed with alumina ceramics. In addition to outstanding mechanical properties, zirconia ceramics have, similar to other surgical grade ceramics, a high biocompatibility and a high resistance to scratching. The radioactivity of zirconia ceramic, which has been the subject of contradictory data, now is well understood and managed with appropriate standards. The long-term stability of zirconia ceramics recently has been studied extensively and precise models allow a good prediction of their long-term behavior. In vitro wear tests against polyethylene and clinical data confirm the low wear rate associated with the use of ceramic femoral heads. The use of zirconia femoral heads in ceramic-on-ceramic total hip prostheses also has been investigated and now is clinically effective. PMID- 11039798 TI - Wear of alumina-on-alumina total hip arthroplasties at a mean 11-year followup. AB - The surface topography of 11 alumina-on-alumina hip arthroplasties retrieved for aseptic loosening at a mean 11-year followup was investigated. Macroscopic wear was assessed using a coordinate measuring machine. Microscopic wear features were evaluated by Talysurf analysis. Scanning electron microscopy was used to look at the alumina microstructure. Components were classified into three groups: (1) low wear with no sign of wear and average arithmetic roughness values below 0.05 microm; (2) stripe wear with a visible oblong worn area on the femoral heads and penetration rates below 10 microm/year; and (3) severe wear with a visible loss of material on both components, showing total roughness values as much as 4 microm and maximum penetrations higher than 150 microm. Alumina quality assessed by grain size measurements and porosity percentages improved progressively from 1977 to 1988. This resulted in a correlated decrease of the microscopic wear magnitude. However, on a macroscopic scale, factors responsible for either a load increase (weight, young age, and male gender) or impairment in the load distribution over the component surfaces (large grain size, nonoptimal initial cup inclination, and cup migration and/or tilting) increased the penetration rates. PMID- 11039799 TI - Medium-term results of a modern metal-on-metal system in total hip replacement. AB - Since 1988, metal-on-metal articulation from cobalt-chromium-molybdenum alloy was reintroduced into hip arthroplasty as an alternative to metal-on-polyethylene or ceramic-on-polyethylene components. Modular joint surfaces were developed for the second generation metal-on-metal articulation using newly introduced and proven prosthetic implants. Since 1990, 78 patients with 78 uncemented total hip replacements were followed up in a prospective study. The mean followup was 60 months. Three patients were lost to followup. The average age of the patients at the time of surgery was 48.8 years. Thirty-three patients had been operated on previously. No early infections occurred; one late infection occurred after 3 years. Dislocation of the prosthesis occurred in one patient who was lost to followup. In two patients ectopic ossifications were removed 17 and 27 months postoperatively. At revision surgery no metallosis could be identified. At the last followup examination, the Harris hip score was 96.8 points on average. There was no evidence that the metal-on-metal articulation gave rise to new problems or complications. Metal-on-metal articulation reduced wear considerably in the authors' previous experience. It is hoped that foreign body reactions are reduced significantly so that an alternative for total hip replacement in younger and active patients will be available. PMID- 11039800 TI - Press-fit metal-backed alumina sockets: a minimum 5-year followup study. AB - Two hundred thirty-four consecutive alumina-on-alumina hip replacements using a press-fit metal-backed socket, performed on 214 patients (98 women, 116 men) have been reviewed. These included 201 primary procedures and 33 revision procedures. The median age of the patients at the time of surgery was 62 years (range, 21-83 years). Fourteen patients (16 hips) died from unrelated causes. Eleven patients (11 hips) underwent a total hip arthroplasty revision for recurrent dislocation (one hip), deep infection (two hips), fracture of alumina femoral head (one hip), persistent hip pain (one hip) and aseptic loosening (six hips). The survival rate after 9 years was 93.4% when revision of the prosthesis was considered the end point, and 97.4% if revision of the prosthesis for aseptic loosening was considered the end point. Results were assessed in the surviving patients with a minimal 5-year followup (170 patients, 184 hips). At the median followup of 7.8 years, the average Merle d'Aubigne and Postel score had improved from 11.9- to 17.7. Results were graded as excellent in 148 hips (80.5%), very good in 31 hips (17%), good in two hips (1%), and fair in three hips (1.5%). Radiologic data were documented for 134 patients (143 hips). Three sockets (2%) had a complete and nonprogressive radiolucent line less than 1-mm thick, one stem (0.7%) had lucencies involving five zones, and two stems (1.4%) had isolated femoral osteolysis. Neither component migration nor acetabular osteolysis were detected. A press-fit metal-backed socket may offer a good solution for alumina socket fixation when combined with a careful surgical technique of implantation. PMID- 11039801 TI - Video-assisted thoracoscopic surgery in managing tuberculous spondylitis. AB - The literature includes no studies on the use of video-assisted thoracoscopic surgery in the management of tuberculous spondylitis, and its role in the management of tuberculosis involving the thoracic spine remains unclear. The authors experience with 10 consecutive patients (six women, four men) who underwent video-assisted thoracoscopic surgery for the treatment of spinal tuberculosis involving levels from T5 to T11, from January 1996 to December 1997, was analyzed. Using the extended manipulating channel method (2.5-3.5 cm portal incisions), video-assisted thoracoscopic surgery was performed with a three portal technique (seven patients) or a modified two-portal minithoracotomy technique that required a small incision for the thoracoscope and a larger incision, measuring 5 to 6 cm, for the procedures in three patients. All the patients were studied prospectively. The followup ranged from 17 to 42 months (mean, 24 months). Postoperative complications included one lung atelectasis. Pleural adhesions, owing to local inflammation or paravertebral abscess, were seen in four patients and one patient with severe pleurodesis needed an open technique for treatment. Postoperative air leaks were seen in four (40%) of 10 patients but all were transient. The average neurologic recovery was 1.1 grades on the Frankel's scale. The data from this series of patients with tuberculous spondylitis show that video-assisted thoracoscopic surgery has diagnostic and therapeutic roles in the management of tuberculous spondylitis. Technically, a combination of thoracoscopy and conventional spinal instruments to perform video assisted thoracoscopic surgery through the extended manipulating channels, which were placed slightly more posterior than usual, was effective and safe. PMID- 11039802 TI - Pain and orthopaedic and neurologic signs after lumbar discectomy: a 2-year followup. AB - In a prospective study of 161 consecutive patients with lumbar discectomy, pain, lumbar mobility, and neurologic and root tension signs were followed up for at least 2 years. Sciatica and root tension signs decreased promptly after surgery and remained largely unchanged during followup, which was not the case for neurologic signs. Similarly, pain relief was not associated with neurologic signs but was associated with lumbar mobility and root tension signs. Patients without neurologic symptoms before surgery did not report more sciatica after 2 years than did those with positive neurologic signs before surgery. Positive crossed Lasegue sign and restricted lumbar mobility before surgery predicted better chances for postoperative pain relief. Patients with a ruptured anulus fibrosus at surgery had less sciatica and back pain after surgery than did patients with an intact anulus fibrosus. PMID- 11039803 TI - Total hip arthroplasty in patients receiving Workers' Compensation. AB - Numerous studies have reported on the adverse outcome of patients who sustain job related injuries. In addition, studies have reported poor outcomes in patients receiving Workers' Compensation who undergo elective surgery. This study sought to determine the influence of Workers' Compensation on the outcome of patients who had undergone primary total hip arthroplasty. Between January 1984 and December 1996, 44 patients (48 hips) were studied. Of these, 17 were men and five were women with a mean age of 45 years (range, 27-76 years) at the time of surgery. These patients were receiving compensation benefits and were matched directly with a group of 22 patients who had 24 arthroplasties and were not receiving compensation. After a mean final followup of 77 months (range, 25-125 months), the compensation group had a mean Harris hip score of 86 points (range, 54-95 points). The matched control group had a mean Harris hip score of 92 points (range, 79-100 points) at a mean final duration of followup of 80 months. Two patients (9%) had undergone revision surgery for aseptic loosening at 28 and 67 months. The percentage of patients with good or excellent results did not differ significantly between the two groups. Based on these findings, the authors think that Workers' Compensation does not negatively influence the outcome of total hip arthroplasty. PMID- 11039804 TI - Bipolar versus total hip arthroplasty for hip osteonecrosis in the same patient. AB - The authors studied 28 patients with bilateral avascular necrosis of the femoral head who were treated with a cementless bipolar endoprosthesis in one hip and cementless total hip arthroplasty in the other. All the hips selected for bipolar endoprostheses were classified as having avascular necrosis of the femoral head Ficat Stage III, and all the hips selected for total hip arthroplasty were classified as having Ficat Stage IV avascular necrosis. After a midterm followup of an average of 6.4 years (range, 4-12 years), 24 of 28 hips that received bipolar endoprostheses were considered satisfactory, whereas 23 of 28 hips in which an arthroplasty was done were considered satisfactory. After a followup of more than 6 years, the cartilaginous space of the acetabulum could be preserved in 25 hips (89.3%) that received a bipolar endoprosthesis. There were no statistical differences in both groups in terms of clinical result, thigh pain, groin pain, osteolysis, dislocation, and revision rate. Total hip arthroplasty is not the preferred treatment for all patients with hip osteonecrosis. In young patients with Ficat Stage III osteonecrosis with Grade 0 or Grade I cartilage, the use of a cementless bipolar endoprosthesis with a bone ingrowth stem may be considered as an alternative to total hip arthroplasty. PMID- 11039805 TI - The safety and efficacy of intraoperative heparin in total hip arthroplasty. AB - A single dose of unfractionated heparin (15 U/kg), administered intravenously before surgery on the femur suppresses thrombogenesis during total hip replacement. Nine hundred eighty-nine patients (1021 hips) who received one dose of intraoperative heparin with hypotensive epidural anesthesia were followed up prospectively for 3 months. Asymptomatic deep vein thrombosis assessed by ultrasound in the first 198 consecutive patients showed an incidence of 7.1% (14 of 198). The incidence of clinical deep vein thrombosis in the subsequent 791 patients was 0.88% (seven of 791). Symptomatic pulmonary embolism occurred in 0.5% (five of 989). No patients died and there was one major bleeding episode. Based on this favorable experience, intraoperative heparin appears safe and efficacious as thromboembolic prophylaxis. PMID- 11039806 TI - Osteolytic indicators found in total knee arthroplasty synovial fluid aspirates. AB - Interleukin-1beta and tartrate resistant acid phosphatase concentrations in synovial fluid aspirates were examined to determine if they could be used as indicators of increased synovial inflammation and an osteolytic reaction in patients having total knee arthroplasty. Synovial aspirates were obtained from seven patients with severely osteoarthritic knees that were scheduled for primary total knee arthroplasty and from 20 patients with knees scheduled for total knee arthroplasty revision. Eleven of the revision cases involved titanium alloy prostheses and nine involved cobalt chrome alloy prostheses. The interleukin 1beta and tartrate resistant acid phosphatase concentrations were obtained and compared between the group having primary total knee arthroplasty and the group having revision total knee arthroplasty. The knees having revision surgery had higher concentrations of interleukin-1beta and tartrate resistant acid phosphatase than did the knees having primary total knee arthroplasty. These results indicate a greater inflammatory and osteolytic response in knees having revision surgery. Although the osteoarthritic knees and the knees needing revision surgery in this study are considered to have an inflammatory state, it was only after total knee arthroplasty when particulate wear debris would be present that appreciable concentrations of interleukin-1beta and tartrate resistant acid phosphatase were produced. PMID- 11039807 TI - Rotating hinge total knee arthroplasty in severly affected knees. AB - A consecutive series of 24 knees in 21 patients who received a Finn rotating hinge for primary (nine knees) or revision (15 knees) total knee arthroplasty between August 1993 and January 1997 was reviewed. The average followup was 33 months (range, 21-62 months) for all patients in the study. Seventeen patients (20 knees) were followed up for more than 2 years. Twenty-four knees (21 patients) were categorized according to Knee Society scoring criteria: 37.5% (nine knees) were Category A, 25% (six knees) were Category B, and 37.5% (nine knees) were Category C. Using the Knee Society knee and function scores, clinical and radiographic results were assessed and outcome analysis was determined. The average Knee Society knee score improved from 44 points (range, 5-64 points) before surgery to 83 points (range, 45-95 points) after surgery; the average functional score according to the Knee Society system improved from 10 points (range, 0-35 points) before surgery to 45 points (range, 0-100 points) after surgery. Pain and function markedly improved after surgery. For treatment of the most severely affected knees with compromised bone and ligamentous instability, the Finn total knee replacement appears to be an acceptable option. As a rotating hinge design, the prosthesis at early followup provides excellent pain relief, restoration of walking capacity, and stabilization, without evidence of early mechanical failure. PMID- 11039808 TI - Distal soft tissue procedure and proximal metatarsal osteotomy in hallux valgus. AB - The results of a distal soft tissue procedure and a proximal metatarsal osteotomy in patients with symptomatic hallux valgus deformity were reviewed. The series consisted of 33 patients (47 feet; mean age of patients, 44 years). The average followup period was 48 months. At followup, 41 feet (29 patients, 85%) were free from pain at the first metatarsophalangeal joint. In six feet (four patients), the pain was improved but persisted. The mean hallux valgus angle was 38 degrees before surgery and 13.8 degrees after surgery. The mean intermetatarsal angle was 17.7 degrees before surgery and 7 degrees after surgery. The postoperative hallux valgus angle and intermetatarsal angle in patients who had pain at the first metatarsophalangeal joint after surgery were greater than those in patients without pain after surgery. This procedure corrects the hallux valgus deformity and relieves the symptoms, but careful attention should be paid to the surgical technique to obtain consistent and satisfactory results. PMID- 11039809 TI - Evaluation of pathologic abnormalities of clubfoot by magnetic resonance imaging. AB - Three-dimensional analyses of clubfoot in infants younger than 1 year of age were done using magnetic resonance imaging in an in vivo study. Twenty-one patients (31 feet) with congenital clubfoot were examined. The average age at examination was 8.6 months (range, 4-12 months). All patients originally were treated using corrective casts. Seventeen feet required complete subtalar release operations and the remaining 14 feet were treated conservatively with various orthoses. Four measurements using magnetic resonance imaging were performed in the transverse and coronal planes as follows: the calcaneus adduction angle, to define the degree of medial rotation of the calcaneus in the transverse plane; the navicular angle, to define the degree of medial displacement of the navicular; the talus neck angle, to define the degree of medial angulation of the talus; and the calcaneus shift index, to define the degree of medial shift of the calcaneus beneath the talar head in the coronal plane. In the results, all four measurements of clubfoot on magnetic resonance imaging were statistically different from those of normal feet. In the surgical group there were statistical differences in the calcaneus adduction angle, the navicular angle, and the calcaneus shift index (including two feet of patients whose parents had rejected proposed treatment), compared with the conservative group, but there was no statistical difference in the talus neck angle. Magnetic resonance imaging could delineate the three-dimensional abnormalities of the tarsal bones in clubfoot and quantitatively evaluate the severity of clubfoot. PMID- 11039810 TI - Arthroscopic treatment for localized pigmented villonodular synovitis of the knee. AB - This study investigated 11 patients with localized pigmented villonodular synovitis of the knee that was diagnosed and treated by arthroscopic technique. There were six male and five female patients between the ages of 15 and 59 years (mean, 34.6 years). Seven patients reported extension limitation without joint line tenderness. Four of the 11 patients had a history of trauma before the onset of knee symptoms. All patients were treated by arthroscopic resection with partial synovectomy. The most common involved site was the anteromedial synovium near the anterior horn of the medial meniscus (five patients). The remaining cases were located in the anterior fat pad (two patients), suprapatellar pouch, posteromedial compartment, medial gutter, and the anterior horn of the lateral meniscus. Nine patients had one mass, and the remaining patients each had two or three masses. There was no evidence of recurrence at followup for an average of 29.9 months (range, 24-48 months). Arthroscopy is effective in the diagnosis of localized pigmented villonodular synovitis with minimal morbidity, and complete arthroscopic excision can be considered the definitive treatment for localized pigmented villonodular synovitis. PMID- 11039811 TI - Langerhans' cell histiocytosis in patients older than 21 years. AB - In this retrospective review of 541 patients with Langerhans' cell histiocytosis, 211 (39%) patients were older than 21 years of age, whereas 330 (61%) were younger than 21 years of age. The adult patients had a mean age of 32 years (range, 21-69 years) with 159 (75%) men and 52 (25%) women, whereas the pediatric patients consisted of 176 (55%) boys and 144 (45%) girls. This male predominance in adults was statistically significant. Three adults had the Hand-Schuller Christian variant, whereas the remaining adults (208) had eosinophilic granuloma. The rib accounted for 25% of the adult lesions and only 8% of the pediatric lesions. Spine involvement was less common in the adult group (3% versus 10%) and was predominantly thoracic. The adult patients had 40 (77%) diaphyseal lesions, 12 (23%) metaphyseal lesions, and no epiphyseal lesions. The pediatric patients had 75 (54%) diaphyseal, 59 (42%) metaphyseal, and five (4%) epiphyseal lesions. Radiographic evaluation revealed similar margin and matrix patterns in both groups, with a geographic lesion without sclerotic borders being the most common pattern. Langerhans' cell histiocytosis is considered a pediatric disease. However, this study showed a significant number (39%) of patients older than 21 years of age with this condition. PMID- 11039812 TI - Anatomic consideration for lumbar percutaneous interbody fusion. AB - The anatomy of the intervertebral foramina L1-S1 was studied by the dissection of 106 foramina levels in 13 human anatomic spine specimens. Twenty foramina were examined in each level from L1 to L5 and 26 foramina from L5 to S1. It appears a safe working canal diameter is 8 mm for L1-L4 and 7 mm for L4-S1. PMID- 11039813 TI - Implantation of a nerve ending into a vein. AB - Neuromas can be painful and physically and psychologically disabling. Among the many methods of treatment available, translocation away from noxious stimuli, such as implantation in muscle or bone, has been used. Veins are easily available and accessible to nerves. The outcome of a nerve ending implanted into a vein and the effects of the implanted nerve on the vein have not been studied. This study evaluated neuroma prevention with a new technique that involves implanting a nerve ending into a vein. The medial branches of the right femoral nerve of 20 rats were transected at midthigh level. The distal segments were excised. The proximal segments were implanted into the femoral vein. The epineurium was sutured to the tunica adventitia using interrupted 10-0 Ethilon sutures. The left side served as controls where the proximal nerve stumps were left lying in their beds. The animals were sacrificed 12 weeks after the operation. Histologic analysis of 12 controls showed neuromas. Twelve implanted nerves showed neuromas that were smaller than those of the controls. The difference was statistically significant. The regenerated nerve fibers were found within the adventitia and muscular wall of the vein. There was no extension of nerve tissue into the vessel lumen and no thrombosis of the vein. Electron microscopic study on the other eight rats showed abnormal large myelinated fibers in the controls and in the implanted nerves. The fibers in the implanted group were degenerated. PMID- 11039814 TI - Bone bonding ability of a new biodegradable composite for internal fixation of bone fractures. AB - Hydroxyapatite particles and poly(L-lactide) composites for internal fixation of bone fractures have been developed based on the hypothesis that incorporation of hydroxyapatite particles in a poly(L-lactide) matrix might enhance bone bonding. This study evaluated the bone bonding ability of these biodegradable composites. Two types of hydroxyapatite and poly(L-lactide) composite were used in this study: calcined hydroxyapatite/poly(L-lactide) and uncalcined hydroxyapatite/poly(L-lactide). Rectangular plates (2 x 10 x 15 mm) of each composite or poly(L-lactide) were implanted into the metaphysis of the tibiae of 33 male rabbits, and the failure load was measured by conducting a detaching test 8, 16, and 25 weeks after implantation. The failure loads of calcined hydroxyapatite/poly(L-lactide), uncalcined hydroxyapatite/poly(L-lactide), and poly(L-lactide), respectively, were 13.60, 13.95, and 0.46 N at 8 weeks; 29.84, 24.09, and 2.86 N at 16 weeks; and 25.50, 29.67, and 2.43 N at 25 weeks. Histologic observation revealed that the composites formed direct contact with the bone. The results in this study indicate that the composites improved the strength of the interface between bone and plate. This improved interfacial strength lead to a substantial decrease in the frequency of implant loosening in the treatment of fractured bones by internal fixation. PMID- 11039815 TI - The adaptation of perimuscular connective tissue during distraction osteogenesis. AB - It is not known whether the decreased range of motion observed during distraction osteogenesis results from the lack of adaptation of muscle or from fibrosis in the perimysium. The adaptation of the perimysium in the tibialis anterior muscle in skeletally immature rabbits using two distraction regimens (0.7 and 1.4 mm/day with 15% lengthening) was characterized. The resulting data indicate that during distraction osteogenesis, the muscle adapts by reorganization of its connective tissue. At a lengthening rate of 1.4 mm/day, there is perimysial fibrosis without major cellular pathologic abnormalities in the muscle fibers. The increase in perimysial thickness is characterized by an increase of collagen Type I. In addition, collagen Type I is deposited around the endomysium. The increase in total collagen and its cross-linking are dependent on the lengthening rate. The faster lengthening rate also leads to a significant decreased passive plantar flexion. Supplemental growth of the tibia was not observed, and a lack of adaptation in the muscle (based on resting length) was not seen. Together, the data suggest that decreased range of motion during distraction osteogenesis may be a function of the adaptation of the perimysium rather than of the muscle fibers. PMID- 11039816 TI - Effects of reconstructed radial collateral ligament on index finger mechanics. AB - Twenty fresh frozen hand specimens from cadavers were studied. Physiologic levels of extrinsic muscle loads were applied to the extrinsic flexor tendons of the index finger to simulate tip pinch of the finger on a fixed plate. The acute effects of transection of the radial collateral ligament and accessory radial collateral ligament (radial collateral ligament complex) with and without transection of the dorsal capsule and volar plate on the position of the proximal phalanx with respect to the metacarpal bone of the index finger were investigated. The acute effects of reconstruction of the radial collateral ligament, for each of two different surgical techniques, on the position of the proximal phalanx also were investigated. The spatial positions of the metacarpal bone and proximal phalanx were measured with a six-degree-of-freedom digitizing system for flexion angles from 0 degrees to 90 degrees in increments of 15 degrees. Transection of the radial collateral ligament complex resulted in significant increases in ulnar deviation (adduction) of the proximal phalanx and in volar translation. Additional transection of the dorsal capsule and volar plate caused significant increases in ulnar deviation, pronation, volar translation, and ulnar shift. The first surgical technique, one traditionally used to reconstruct the metacarpophalangeal joint of the thumb, failed to return the three-dimensional position of the proximal phalanx on the metacarpal head of the index finger to normal. The second surgical technique, based on anatomy, returned the position of the proximal phalanx to levels not statistically different from normal for most flexion angles. PMID- 11039817 TI - Recurrent arm pain in a 77-year-old man. PMID- 11039819 TI - Telemedicine for the new century. PMID- 11039818 TI - Antiviral treatment of HIV infection. PMID- 11039820 TI - Cat allergen levels in public places in New Zealand. AB - AIMS: Cat allergen (Fel d 1) is a known risk factor for asthma. Studies have demonstrated Fel d 1 in both public buildings and domestic dwellings where cats have never been. The aims of this study were to measure reservoir Fel d 1 levels in public buildings in New Zealand, to examine determinants of these levels and to compare them with previously measured domestic levels. METHODS: Dust was obtained in two centres (Wellington and Christchurch) from hotels, hospitals, rest homes, churches, primary schools, childcare centres, cinemas, bank head offices and aeroplanes; and from North Island ski lodges. Measurements of temperature and relative humidity were taken. Information was collected on building characteristics. Fel d 1 levels (microg/g of fine dust) for floors (n=203), beds (n=64) and seats (n=24) were expressed as geometric means (95% confidence intervals). RESULTS: Detectable Fel d 1 levels were found in 95% of floor samples, 91% of bed samples and 100% of seat samples. Fel d 1 levels [geometric mean (95% confidence intervals)] were significantly higher on cinema and domestic aircraft seats [36.8 (20.8-65.3) microg/g and 33.3 (28.0-39.7) microg/g respectively] than on floors [3.6 (2.5-5.1) microg/g and 2.4 (1.8-3.0) microg/g respectively]. Floor Fel d 1 levels in the public buildings sampled were lower than those of domestic dwellings without cats [0.9 (0.6-1.4) microg/g vs 1.7 (1.2-2.4)] microg/g in Wellington and [2.0 (1.6-2.6) microg/g vs 4.0 (2.7 6.0] microg/g in Christchurch. After controlling for potential confounders, floor Fel d 1 levels were higher with carpeted floors (p<0.001) and lower in banks and hospitals (p<0.001). CONCLUSION: Fel d 1 levels in public buildings are low in New Zealand public places except for cinema and domestic aircraft seats where all but one sample had Fel d 1 levels potentially high enough to precipitate asthma symptoms in sensitised individuals. PMID- 11039821 TI - Preventing falls and fall-related injuries among older people living in institutions: current practice and future opportunities. AB - AIMS: To identify existing falls prevention activities and support for future initiatives among residential institutions for older people. METHODS: A self administered questionnaire was sent to all principal nurses/managers of residential institutions. Information was sought on whether falls were perceived to be a problem in the Auckland region, current falls prevention practice and interest in future prevention initiatives. RESULTS: Falls were perceived to be a problem by over 75% of 175 participating institutions. Assessments of footwear, medication use and environmental audits were the most common prevention strategies employed by over 80% of institutions. Almost 70% of institutions indicated their willingness to participate in future prevention projects. CONCLUSION: The current use of fall prevention strategies in institutions is encouraging. However, the strategies that are being employed are not consistent with current evidence about effectiveness. Increased use of vitamin D and possibly calcium supplementation needs to be encouraged as does the use of hip protectors and lower extremity strength training. PMID- 11039822 TI - Health effects of occupational pentachlorophenol exposure in timber sawmill employees: a preliminary study. AB - AIMS: To study the health effects of pentachlorophenol (PCP) exposure in the timber sawmill industry. METHOD: A questionnaire-based, non-random survey was undertaken amongst a group of current and ex-workers who had identified their health concerns as being related to PCP exposure. RESULTS: Low, medium and high exposure groups were identified. A significant dose-response was found between past exposure to Pentachlorophenol and reported symptoms of fever/sweating (47% in the high exposure group), weight loss (33% in the high exposure group), persisting fatigue (74% in the high exposure group), nausea (40% in the medium and high exposure groups) and responses to a screening test for neuropsychological dysfunction (Questionnaire 16) (81% in the high exposure group). No associations were observed with other chronic diseases, apart from emphysema and chronic bronchitis. CONCLUSIONS: This study is based on a self selected sample of PCP-exposed workers whose precise exposure levels are unclear. Thus the findings presented should be regarded as preliminary. Nevertheless, they support clinical experiences and point to the need for further investigation. PMID- 11039823 TI - Ethnic differences in parent/infant co-sleeping practices in New Zealand. AB - AIM: This study was designed to monitor changes in the prevalence of risk factors for sudden infant death syndrome (SIDS) in the New Zealand population. The behaviour of interest is parent/infant co-sleeping. This paper reports parent/infant co-sleeping arrangements of different ethnic groups in New Zealand. METHODS: A stratified random sample of 6268 infants attending Plunket clinics for their three and six-month visits was taken over the years 1995-1996. Maori and Pacific infants were oversampled. Parents who shared a bed with their infant were asked how they arranged the babies sleeping place according to pre-coded diagrams. Routine parent/infant co-sleeping was defined as "bed sharing at least four nights over the last two weeks". RESULTS: There were 2693 infants who shared the bed with their sleeping parents during at least one of the previous 14 nights. Of these infants, 1060 routinely shared the parents' bed. At three months, 56% of routinely co-sleeping infants slept directly in the bed, 29% slept in a raised position, 3% slept in a carrycot or basket, and 5% in other positions. At six months, 60% of the routinely co-sleeping infants slept directly in the bed with their parents, 23% slept in a raised position, 1% slept in a carrycot or basket, and 7% in other positions. There were significant differences in the co-sleeping locations by ethnicity. CONCLUSION: There is still some ongoing dispute as to whether parent/infant co-sleeping is a risk factor for SIDS. This study has identified differences in the way infants co-sleep with their parents and this can be used to clarify infant care practices in relation to SIDS. PMID- 11039824 TI - Cannabis policy: issues and options. AB - This paper examines some of the key points relevant to the debate about cannabis policy in New Zealand. It provides a brief overview of cannabis use patterns, the cannabis market and the public health implications of use. It describes the various strategies which comprise our current cannabis policy and the context of and primary concerns relating to it. Several alternative policy options are explained and in each case some evaluative comments are made. These alternatives are: total prohibition with an expediency principle; prohibition with civil penalties, partial prohibition, and regulation of private enterprise producers/distributors. From a public health perspective none of these options is unproblematic but each has the potential to overcome some of the disadvantages of the current policy and each needs to be further evaluated. PMID- 11039825 TI - Ophthalmia neonatorum in New Zealand. PMID- 11039826 TI - Breast cancer screening policy. PMID- 11039827 TI - SIDS and the toxic gas theory. PMID- 11039828 TI - Is mammography such a pain? PMID- 11039829 TI - Privacy of patient records. PMID- 11039830 TI - Airway of babies in car seats. PMID- 11039831 TI - A farewell to GP-obstetrics. PMID- 11039832 TI - Zoonoses. PMID- 11039833 TI - Chemical poisoning and occupational asthma: diagnosis and/or acceptance for current compensation. PMID- 11039834 TI - Hypersensitivity to members of the botanical order Fabales (legumes). AB - Legumes are an important source of proteins and their consumption is very frequent in the Mediterranean region and in some Asian and African countries. In some of these regions, lentils and chickpeas are one of the main food allergens. Legumes are also used as food additives due to their emulsifying properties and can be present in many manufactured foods. These hidden food allergens have the potential of causing adverse reactions in legume-sensitive subjects. The allergenic composition of various legumes has been investigated. They have been found to contain multiple allergens, a few of which have been cloned and sequenced. Legumes contain acid-resistant and thermostable allergens. There is a significant degree of cross-reactivity among legumes, the clinical relevance of which seems to be dependent on the dietary habits in different communities. In Spain, the consumption of several legumes is frequent and, therefore, clinical allergy to more than one species in children is common. Clinical manifestations include cutaneous, digestive and respiratory symptoms. Legumes can cause life threatening reactions in sensitized individuals. Inhalation of steam, powder or flour from some legumes may cause respiratory diseases such as rhinitis, asthma and hypersensitivity pneumonitis. Soybean allergy is generally transitory, but clinical allergy to peanuts is rarely outgrown. The natural history of other legume allergies is less known and more studies are necessary to reach definite conclusions. PMID- 11039835 TI - Personal pollen exposure compared to stationary measurements. AB - The aim of this study was to examine to what extent stationary outdoor pollen measurements are representative for estimating personal exposure to pollen. Ten subjects were studied during a total of 36 days in spring and summer Pollen was sampled using personal SKC total dust samplers and stationary Burkard pollen traps. The personal activity pattern was recorded quarter-hourly as well as the time spent outdoors. As a reference, SKC and Burkard samplers were run stationary and in parallel. Stationary comparison of the samplers showed good correlation (r = 0.981, p <0.001). However, the SKC sampler collected systematically about four times less pollen than the Burkard sampler. Taking into account the systematic difference between the sampling devices, the personal exposure data were about 30% of the stationary pollen concentrations with significant correlation (log transformed data, r = 0.719, p <0.0001). Considering the average time the subjects spent outdoors (14% of sampling time), the indoor-outdoor ratio for pollen was 0.2. In conclusion, pollen reports are reliable for estimating personal exposure over a limited time period although personal pollen exposure is much lower. PMID- 11039836 TI - Cat allergen sampling by a new personal collector (Partrap FA 52). AB - Recent studies carried out by us and others have demonstrated that Fel d 1, the main cat allergen, may be passively transferred by human clothing in cat-free environments. Consequently, the monitoring of the Fel d 1 levels either in indoor environments or on allergen-contaminated clothes of sensitized cat owners should be considered an important tool in prevention strategies. The aim of this study was to evaluate the efficacy of a personal air sampler (Partrap FA 52) in capturing cat allergen from wool fabrics. Seven identical wool webs (80 x 100 cm) were put in the baskets of seven male cats for 1 week. In our laboratory each web was divided into two parts (80 x 50 cm), the first of which was then divided in two parts (40 x 50 cm) and each was vacuumed directly by one collector. The second part was dry-cleaned at a professional cleaners, divided in two parts and then vacuumed. For the dust collection from wool webs we used a fixed high volume air sampler (CF/20 Gelaire Flow Labs, Milan, Italy) and a personal collector (Partrap FA 52, Coppa Biella, Italy). Fel d 1 content was determined using a two site ELISA (ALK-Abello Group, Madrid, Spain). Both air samplers collected cat allergens from cat-exposed wool fabrics before and after dry cleaning. There were significant differences between the levels of Fel d 1 before and after dry cleaning by using either CF/20 or Partrap FA52 and between the levels of Fel d 1 before dry cleaning using CF/20 and Partrap FA 52. The results of our study suggest that Partrap FA 52, although its air flow is half that of the CF/20, is able to collect even residual amounts of cat allergen from wool webs after dry cleaning and consequently may constitute a simple and effective means of monitoring the levels of Fel d 1 on the clothes of cat owners. PMID- 11039837 TI - Complement activation in Brazilian patients with preeclampsia. AB - The aim of this study was to evaluate the complement activation in Brazilian patients with preeclampsia and to correlate it with the severity and clinical outcome of the disease. Plasma levels of C3d, SC5b-9, C3 and C4 were measured in 16 patients with preeclampsia and in 17 normotensive pregnant women. Ten patients developed severe and six mild disease. C3 and C4 levels were determined by turbidimetry using polyclonal specific antisera. C3d concentrations were evaluated through double-decker rocket immunoelectrophoresis and SC5b-9 was assayed by a double-antibody enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody against a neoantigen expressed in the formed complex. The mean levels of all variables were significantly higher in the preeclamptic group (before the delivery) when compared to the normal pregnancies. The complex SC5b-9 followed by C3d showed the most significant results for those comparisons (p < or = 0.00001). The levels of all parameters in the preeclampsia patients decreased significantly after the delivery. Again, the complex SC5b-9 and C3d showed the most significant results (p < or = 0.0004). None of the studied variables showed statistically significant differences regarding the severity of preeclampsia. These results confirm the activation of complement in preeclampsia, suggesting that this activation is related to the disease manifestation. Our findings further emphasize the involvement of complement activation in the pathological manifestations of preeclampsia. PMID- 11039838 TI - Appraisal of latex glove proteins in the induction of sensitivity to multiple latex allergens. AB - Six Hevea brasiliensis latex protein allergens, Hevb 1, Hev b 2, Hev b 3, Hev b 4, and two variants of Hev b 7 (7b and 7c), were purified from Hevea latex, while a seventh protein, Hev b 5, was prepared in recombinant form. The presence of these proteins in glove extracts was indicated by their respective antibodies in the serum of rabbits immunized against the extracts. The relative propensities of IgE binding to the individual latex allergens were compared using sera from latex allergic patients. IgE recognition of Hev b 4, Hev b 7b, Hev b 5 and Hev b 2 was most frequently encountered, with 75, 61, 31 and 28%, respectively, of the patient sera reacting. Sensitivity to multiple latex proteins was common, and out of the 31 seropositive patients, 23 (74%/ ) had IgE against at least two latex allergens, while 12 (39%) had IgE specific for at least three allergens. Statistical analysis of the data suggested that many patients might have acquired sensitivity to Hev b 2, Hev b 4 and Hev b 7b from a common source. (e.g., from latex products). On the other hand, sensitivity to Hev b 5 and to Hev b 7c were interrelated. It is plausible that sensitivity to these two proteins might have been acquired from sources other than latex products (e.g., from certain foods). PMID- 11039839 TI - Bee venom induces high histamine or high leukotriene C4 release in skin of sensitized beekeepers. AB - Histamine is the principal mediator released in the skin during immediate bee venom allergy but the significance of cysteinyl leukotrienes in these reactions is not known. We measured skin histamine and cysteinyl leukotriene release induced by bee venom in six sensitized beekeepers with the skin microdialysis technique. The skin was dialyzed for 2 h after skin prick test with bee venom, and the release of histamine and leukotriene C4 (LTC4) into the microdialysis fractions was measured. Leukotriene E4 (LTE) and methylhistamine excretion into the urine was assayed and whole blood histamine release test was performed. The release of histamine in the skin was variable: either high delayed, high immediate and delayed, weak release or no marked release. The histamine releasability in the skin correlated with that in whole blood. The three subjects with low histamine release exhibited high LTC4 release in the skin as well as high LTE4 excretion into the urine. Thus, the histamine and LTC4 releases were inversely associated with each other. These differences may explain the variation in the clinical reaction by bee stings in sensitized beekeepers. PMID- 11039840 TI - Factors influencing the clinical picture and the differential sensitization to house dust mites and storage mites. AB - Sensitization to house dust mites and storage mites has been studied in a number of papers, but several environmental factors and clinical conditions that differently affect sensitization to these mites are still controversial. The purpose of this study was to determine the influence of climatic conditions, occupation and patient age in the differential sensitization to house dust and storage mites, and also to search for possible different symptoms caused by each group of mites. Eighty patients sensitized to mites but not to other inhalant allergens were studied by case history and by skin prick test and serum IgE to Dermatophagoides pteronyssinus. Dermatophagoides farinae, Lepidoglyphus destructor and Tyrophagus putrescentiae. Home conditions, including content of the allergens Der p 1, Der f 1, Lep d 2 and Tp, were determined for all patients. Human activities, such as farming or similar occupations, and humidity are conditions for preferential sensitization to storage mites, while we found no difference between living in rural or urban areas. Mean age for the onset of sensitization was 6.7 years for house dust mites and 18.7 years for storage mites. Conjunctivitis was more frequent in patients allergic to storage mites, whereas perioral syndrome (itching of the tongue and swelling of the lips) was only seen in patients sensitized to T. putrescentiae. We concluded that climatic and damp conditions and human activity, but not urban or rural living environments, influence the differential sensitization to house dust mites and storage mites. PMID- 11039841 TI - The presence of HLA-DR B1 shared motif does not influence cyclophosphamide and methotrexate cytotoxicity in rheumatoid arthritis patients. AB - In the present study we investigated the relation between cyclophosphamide and methotrexate toxicity and the presence of HLA- DR B1 alleles in rheumatoid arthritis patients. Seventy-eight such patients (67 women and 11 men) were observed for 12 months. Eighteen were treated with intravenous cyclophosphamide, 28 with oral methotrexate, and 32 with intramuscular gold salts. The prevalence of this shared motif was higher in the study population than in the healthy controls. However, detailed observations did not demonstrate a relation between particular genotype and drug intolerance. Based on the obtained findings we concluded that HLA-DR B1 typing cannot affect cyclophosphamide or methotrexate tolerance in rheumatoid arthritis patients. However, taking into account the relatively small number of patients expressing single genotype, further studies are recommended. PMID- 11039842 TI - Allergenic pollen in the subdesert areas of the Iberian peninsula. AB - The yearly distribution of Artemisia and Chenopodiaceae-Amaranthaceae, two of the most common types of pollen in a rural area located in the southeastern part of the Iberian peninsula, was studied over a 3-year period (1995-1997). The particular bioclimatic conditions of the area, such as its subdesert climate, extreme dryness and high mountain location (1,000 m above sea level), have led to the adaptation and abundance of these species in this area. They usually flower in the second half of the year, and are the main pollen types collected in the samples in that time period. The Artemisia pollen levels recorded are the highest in Spain, since there are several species in the area which flower at different times. Chenopodiaceae-Amaranthaceae pollen counts are also very high. The severity of both pollen types was also analyzed. The height of the sampler was taken into account because the quantities at human height can be considerably higher than those recorded at 20 m off the ground. It was concluded that both pollen types should be considered some of the main causes of allergy in this area. PMID- 11039843 TI - Telangiectasia macularis eruptiva perstans presented as a pseudoallergic food reaction. AB - Telangiectasia macularis eruptiva perstans (TMEP) is a rare form of cutaneous mastocytosis observed in less than 1% of cases of mastocytosis. Clinically, anaphylaxis may appear as a result of increased mast cell degranulation in different circumstances. A case of TMEP presented as pseudoallergic reactions to foods is reported in which the appearance of typical lesions on the trunk and their histological analysis together with a negative food allergy study confirmed the diagnosis. PMID- 11039844 TI - Assessment of the metabolic state of skeletal muscles by measurement of glycolytic flux. PMID- 11039845 TI - Alcohol and DNA damage. PMID- 11039846 TI - Hereditary pancreatitis: epidemiology, molecules, mutations, and models. PMID- 11039847 TI - Appropriate use and misuse of blood concentration measurements to quantitate first-pass metabolism. AB - First-pass metabolism commonly is determined from the difference in the area under the blood concentration time curve (AUC) that is observed with oral versus intravenous administration of a compound. It is not fully appreciated that this technique serves as an accurate indicator of first-pass metabolism only when the clearance of the compound under consideration is first order (unsaturated) throughout the range of blood concentrations observed in the study. For example, multiple publications continue to mistakenly use AUC measurements to assess the first-pass metabolism of ethanol, a compound cleared primarily via zero-order kinetics. This report briefly reviews the physiologic basis of measurements of first-pass metabolism, demonstrates the errors that result from application of this technique when clearance is not first order, and, using ethanol as an example, describes a technique that can be used to measure first-pass metabolism when clearance deviates from first order. PMID- 11039848 TI - Glycolysis in the human muscle: a new approach. AB - The flow response time theory allows the global assessment of a metabolic pathway. This study describes the first application of this concept to explore glycolysis on human skeletal muscle extracts. The muscle extract is used to convert glucose or glucose-6-phosphate into glycerol-phosphate through the first part of glycolysis. The functioning of the experimental model is assayed by a continuous recording of the reduced nicotinamide adenine dinucleotide decay in a spectrophotometer. This measurement method was applied to normal and pathologic human skeletal muscles. The aerobic (J(A)) and anaerobic (J(B)) fluxes and the time (t99) needed for the transition from J(A) to J(B) were measured under a wide clinical temperature range (30 degrees C to 40 degrees C). The two studied muscle types (gluteus maximus and tibialis anterior) have similar glycolytic flux values, with an identical functional modality. The thermal response of glycolysis is not linear, with a high thermal sensitivity in the hypothermic range (30 degrees C to 38 degrees C) and a thermal insensitivity in the hyperthermic range (37 degrees C to 40 degrees C). On the same type of muscle (tibialis anterior), a pathologic process can induce different variations in the glycolysis patterns, but further data are needed to clear this point. The flow response time concept allows an accurate assessment of glycolysis in the human skeletal muscle, whether normal or pathologic. This approach is interesting for evaluating the global influence of different stimulations on a metabolic pathway, such as temperature variation. PMID- 11039849 TI - Increased lipid and protein oxidation and DNA damage in patients with chronic alcoholism. AB - Increased oxidative stress has been speculated to be one possible mechanism of ethanol toxicity. This study evaluates malondialdehyde (MDA) and protein carbonyl content in serum as markers of oxidative stress and DNA damage in lymphocytes in the same patients with chronic alcoholism. Patients with chronic alcoholism showed a significant increase in MDA levels and protein carbonyl content of their serum as compared with non-alcoholic control subjects. Increases in endogenous and H2O2-induced DNA damage were also observed in lymphocytes of patients with chronic alcoholism. In addition, there were significant correlations between endogenous and H2O2-induced DNA damage and serum MDA or protein carbonyl content in patients with chronic alcoholism. These results clearly indicate the presence of oxidative stress in patients with chronic alcoholism. PMID- 11039850 TI - Residues F16-G33 and A784-N823 within platelet thrombospondin-1 play a major role in binding human neutrophils: evaluation by two novel binding assays. AB - The thrombospondin-1 (TSP1) structural requirements within its heparin-binding domain (HBD)(30 kd) or within the other domains of the molecule (450 kd) that interact with neutrophils (PMNs) have not been delineated. Synthetic peptides based on the HBD, a TSP1 proteolytic fragment lacking the HBD, a large C-terminal domain of TSP1 (210 kd), a TSP1 recombinant fragment (rTSP1(784-932)), and a monoclonal antibody directed against the TSP1 type 3 repeats (mAb D4.6) were utilized to map such structural requirements on TSP1. Synthetic peptides containing a heparin-binding motif and encompassing residues F16-G33 or A74-S95 of TSP1 competed quantitatively with iodine 125-labeled TSP1 for binding to heparinagarose beads. However, only F16-G33 was a competitor of TSP1 binding to PMNs, suggesting that the sequence F16-G33 within the HBD plays a role in PMN binding. The interaction site within the 450-kd fragment was further narrowed. A TSP1 -derived proteolytic fragment (210 kd), a recombinant TSP1 fragment (rTSP1(784-932)), and a type 3 repeat anti-TSP1 monoclonal antibody (mAb D4.6) competed for the binding of 125I-labeled TSP1 to PMNs. The N-terminal of rTSP1(784-932) and C-terminal sequence analysis of TSP1-210 kd delineated the structural requirements for the second binding region for PMNs-namely, residues A784-N823. PMID- 11039851 TI - Activated platelets adherent to an intact endothelial cell monolayer bind flowing neutrophils and enable them to transfer to the endothelial surface. AB - We have investigated the role that platelets may play in promoting adhesion of neutrophils to morphologically intact endothelium. Immortalized human microvascular endothelial cells (HMEC-1) were grown to confluence in a glass capillary (microslide) and incorporated in a flow-based assay that allowed video microscopic quantitation of adhesive interactions of perfused, isolated neutrophils (wall shear rate 140 s(-1)). Platelets (with or without stimulation with thrombin) were first sedimented onto the HMEC-1 cells and formed discrete attachments covering <1% of the surface area. When neutrophils were perfused over the platelet-treated HMEC-1 cells, many short-lived adhesive interactions were seen (lasting approximately 0.3 seconds), whereas none were seen for monolayers without platelets. Few of these interactions converted to stationary adhesion, and only small numbers of neutrophils remained attached after a period of washout unless they were pre-stimulated with formyl peptide (fMLP; 10(-7) mol/L). Then about 30% of adhesive interactions by activated neutrophils were seen to transform to a stationary adhesion, and numerous adherent cells remained after a period of washout. Studies with function-blocking antibodies showed that capture of the neutrophils was dependent on P-selectin exposed on platelets. Initial immobilization was mediated predominantly by the beta2-integrin CD11b/CD18 expressed by neutrophils, but CD11a/CD18 also appeared to play a role in prolonged attachment. Visually, adhesion first occurred at sites occupied by platelets, but some activated neutrophils migrated onto the endothelial cells. These studies indicate that even small numbers of platelets that have adhered to morphologically intact endothelium have the potential to capture flowing neutrophils and facilitate their immobilization at the vessel wall and so promote inflammatory and thrombotic intercellular interactions. PMID- 11039852 TI - Pregnancy and amyloidosis: II. Suppression of amyloidogenesis during pregnancy. AB - The observation of a deleterious effect of pregnancy on kidney function in amyloidosis of familial Mediterranean fever suggests that pregnancy may enhance amyloidogenesis. To determine whether pregnancy may indeed affect amyloidogenesis, pregnant mice were made amyloidotic by administration of amyloid enhancing factor (AEF) and AgNO3 at different points in time from conception, and amyloid- deposition was studied with the crush-and-smear technique. A possible effect of exogenous female sex hormones (beta-estradiol and progesterone) on amyloidogenesis was studied by administration of these hormones during amyloid induction in nonpregnant female mice. Amyloidogenesis was found to be significantly suppressed in mice during pregnancy. The reduction was possibly related to the effect of pregnancy on the inflammatory stimulus (AgNO3) and not on the administered AEF. Exogenous estrogen and progesterone failed to inhibit amyloidogenesis in nonpregnant mice. These findings suggest that pregnancy may suppress amyloidogenesis in mice. The suppression is caused by an anti inflammatory effect of pregnancy. Estrogen and progesterone are probably unrelated to this finding. PMID- 11039853 TI - Serum soluble Fas (CD95) and Fas ligand profiles in chronic kidney failure. AB - Apoptosis, or programmed cell death, is an active form of cell death that is initiated by a number of stimuli and is intricately regulated. Apoptosis in both excessive and reduced amounts has pathophysiologic implications. Accelerated programmed cell death has been observed in leukocytes among patients with chronic renal failure (CRF). This has been ascribed in part to the retention of uremic toxins. The Fas/Fas ligand (FasL) system is a key regulatory apoptotic pathway. Membrane-bound Fas is a cell-surface receptor that transduces apoptosis after interaction with membrane-bound or soluble FasL (sFasL). By contrast, soluble Fas (sFas) binds sFasL and inhibits its activity. In an attempt to examine the balance between these soluble factors in uremia, we measured soluble sFas and sFasL levels in the serum of healthy control subjects and patients with various degrees of CRF and examined the distribution of the various molecular mass fractions of these proteins in uremic serum. In brief, serum was obtained from 15 healthy volunteers, 17 patients with CRF, 11 patients undergoing maintenance hemodialysis (HD), and 7 patients undergoing peritoneal dialysis (PD). Serum sFas and sFasL were measured by enzyme-linked immunosorbent assay, and their molecular distribution was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by immunoblot. Compared with results in healthy control subjects, sFas levels were significantly higher in patients with CRF and in patients undergoing dialysis. There was a significant inverse correlation between sFas levels and creatinine clearance. Serum sFasL levels were not different among the four groups. However, the sFas-to-sFasL ratio was significantly lower in healthy control subjects as compared with patients with CRF and patients undergoing dialysis. Immunoblots and densitometric analyses of sFas and sFasL depicted a known 48-kd sFas, a known 27-kd sFasL, and a 60-kd sFas-sFasL protein aggregate signal. In conclusion, serum sFas levels are increased in patients with various degrees of CRF and may bind circulating sFasL, thereby minimizing mediation of cellular apoptosis. PMID- 11039854 TI - Toward a developmental operational definition of autism. AB - Traditional approaches to diagnosing autism emphasize delays in communication and socialization. Traditional diagnostic schemes typically list symptoms (e.g., lack of eye contact), but provide little guidance on how to incorporate information about developmental level in making a diagnosis. Because standardized measures of adaptive behavior can provide information about children's communication, socialization, and other behavior relative to their age, they may be useful tools for diagnosing autism. This study investigated the ability of the Vineland Adaptive Behavior Scales to identify children with autism. Vineland scores and measures of intellectual functioning were obtained for children with autism, PDDNOS, and other developmental disorders (DD). Discriminant function analyses indicated that the autism and combined nonautism (PDDNOS and DD) groups could be differentiated on the basis of socialization, daily living skills, and serious maladaptive behaviors. Socialization alone accounted for 48% of the variance in diagnosis. Using regression analyses derived from a large normative sample, adaptive behavior scores were predicted from chronological age (CA) and mental age (MA). Socialization scores in the autism group were substantially below the level predicted from CA or MA. An index derived from the ratio of actual to predicted socialization scores correctly classified 86% of both autism and nonautism cases. Findings suggest that comparison of obtained Vineland socialization scores to those predicted by CA or MA may be useful in clarifying the diagnosis of autism. PMID- 11039855 TI - Social and psychiatric functioning in adolescents with Asperger syndrome compared with conduct disorder. AB - Lack of standardized phenotypic definition has made outcome studies of Asperger syndrome (AS) difficult to interpret. This paper reports psychosocial functioning in 20 male adolescents with AS, defined according to current ICD-10 criteria, and a comparison group of 20 male adolescents with severe conduct disorder. Subjects were gathered from clinical referral. Evaluation used standardized interviewer rated assessments of social functioning and psychiatric morbidity. The AS group showed severe impairments in practical social functioning despite good cognitive ability and lack of significant early language delay. High levels of anxiety and obsessional disorders were found in AS; depression, suicidal ideation, tempers, and defiance in both groups. Results are compared with those from other studies. Relevance to clinical ascertainment and treatment is discussed. PMID- 11039856 TI - Episodic memory and remembering in adults with Asperger syndrome. AB - A group of adults with Asperger syndrome and an IQ-matched control group were compared in remember versus know recognition memory. Word frequency was also manipulated. Both groups showed superior recognition for low-frequency compared with high-frequency words, and in both groups this word frequency effect occurred in remembering, not in knowing. Nor did overall recognition differ between the two groups. However, recognition in the Asperger group was associated with less remembering, and more knowing, than in the control group. Since remembering reflects autonoetic consciousness, which is the hallmark of an episodic memory system, these results show that episodic memory is moderately impaired in individuals with Asperger syndrome even when overall recognition performance is not. PMID- 11039857 TI - Photographic cues do not always facilitate performance on false belief tasks in children with autism. AB - Previous studies have indicated that a pictorial representation of a prior belief can help 3-year-old children (Mitchell & Lacohee, 1991) as well as children with autism (Charman & Lynggaard, 1998) to pass false belief tasks that used the deceptive box or "Smarties" paradigm. The studies reported here attempted to replicate these findings using the unexpected transfer or "Sally-Anne" paradigm, which requires children to predict the actions of a protagonist on the basis of a false belief. Results showed no facilitative effect on "Sally-Anne" task performance for the children with autism or for comparison children of either representational or nonrepresentational cues. This effect was found even in children who benefited from the intervention with the deceptive box paradigm. The findings raise issues regarding the way false belief tasks are conceptualized by experimenters and the demands different false belief paradigms make on children. PMID- 11039858 TI - The children's Social Behavior Questionnaire for milder variants of PDD problems: evaluation of the psychometric characteristics. AB - The Children's Social Behavior Questionnaire (CSBQ) contains items referring to behavior problems seen in children with milder variants of PDD. Data of large samples of children diagnosed as having high-functioning autism, PDDNOS, ADHD, and other child-psychiatric disorders were gathered. Besides the CSBQ, parents completed the Autism Behavior Checklist (ABC) and the Child Behavior Checklist (CBCL). The data provided the basis for scale construction of the CSBQ, a comparison of the CSBQ scales with other instruments and a comparison of groups on scores on the CSBQ. The 5 scales obtained referred to Acting-out behaviors, Social Contact problems, Social Insight problems, Anxious/Rigid behaviors and Stereotypical behaviors. Results show that the CSBQ has good psychometric qualities with respect to both reliability and validity. A comparison of the different groups showed that significant group differences were found on all scales. In general, the autism group received the highest scores, followed by the PDDNOS group and the ADHD group. Exceptions were on the Acting-out scale, where the ADHD group scored highest and on the Social Insight scale, where no significant difference was found between the PDDNOS group and the ADHD group. Implications of the results and suggestions for further research are discussed. PMID- 11039859 TI - The effects of social interactive training on early social communicative skills of children with autism. AB - Growing attention has been directed at the relation between early social communicative skills of children with autism and subsequent development of these children's social and communicative functioning. We reviewed 16 empirical studies that investigated the effects of social interactive interventions designed to increase early social communicative skills of young children with autism by increasing their role as initiator of social interactions. To identify factors relating to treatment effectiveness, we analyzed studies in relation to participant characteristics, settings, target behaviors, training methods, and results. To determine durability of treatment, we analyzed generalization effects across persons, settings, stimuli, and time. Increases were found for social and affective behaviors, nonverbal and verbal communication, eye contact, joint attention, and imitative play. Limited generalization or maintenance of target behaviors was reported. Findings are discussed in relation to critical variables that may relate to treatment effectiveness in future research and practice efforts. PMID- 11039860 TI - Vocal atypicalities of preverbal autistic children. AB - This study was designed to evaluate the nature of early vocal behaviors in young children with autism. Recent methodological and conceptual advances in the study of infant preverbal vocalizations were used to provide a detailed examination of the vocal behavior of young preverbal children with autism and comparison children with developmental delays. Results revealed that children with autism did not have difficulty with the expression of well-formed syllables (i.e., canonical babbling). However, children with autism did display significant impairments in vocal quality (i.e., atypical phonation). Specifically, autistic children produced a greater proportion of syllables with atypical phonation than did comparison children. Consistent with prior reports, the children with autism also displayed a deficit in joint attention behaviors. Furthermore, the atypicalities in the vocal behavior of children with autism appeared to be independent of individual differences in joint attention skill, suggesting that a multiple process model may be needed to describe early social-communication impairments in children with autism. Data are discussed in terms of their implications for future theoretical and applied research, including efforts to enhance the specificity of early diagnostic procedures. PMID- 11039861 TI - The HOPA gene dodecamer duplication is not a significant etiological factor in autism. AB - A recent study has suggested that a dodecamer duplication in the HOPA gene in Xq13 may occur in a significant portion of male patients with autism. We have determined the incidence of this duplication in 202 patients from the South Carolina Autism Study. The incidence of the duplication was not significantly different between patients and controls. Three of the female patients inherited the duplication from nonautistic fathers. In addition, there was no systematic skewing of X inactivation in the female patients with the duplication, or in nonautistic mothers and sisters with the duplication. These findings suggest that the dodecamer duplication in the HOPA gene does not play a significant role in the etiology of autism. PMID- 11039862 TI - Brief report: vocabulary acquisition for children with autism: teacher or computer instruction. AB - This study examined the impact of computers on the vocabulary acquisition of young children with autism. Children's attention, motivation, and learning of words was compared in a behavioral program and an educational software program. The educational software program was designed to parallel the behavioral program, but it added perceptually salient qualities such as interesting sounds and object movement. Children with autism were more attentive, more motivated, and learned more vocabulary in the computer than in the behavioral program. Implications are considered for the development of computer software to teach vocabulary to children who have autism. PMID- 11039863 TI - Siegel's defense: more inaccuracies. PMID- 11039864 TI - The "other" kids with PDD: older, nonverbal, more severely retarded and/or more severely affected individuals with autistic spectrum disorders. PMID- 11039865 TI - Is a large head circumference a sign of autism? PMID- 11039866 TI - Review of segmental and marginal resection of the mandible in patients with oral cancer. AB - This paper reviews the medical literature of the last decade to ascertain the criteria used to assess mandibular invasion by cancer of the oral cavity and to suggest how best to evaluate the mandible with a view to surgical management. It is generally agreed that patients with mandibular invasion should be treated surgically, but the extent of mandibular resection required remains a controversial matter and the accurate preoperative determination of neoplastic invasion of the mandible remains a challenge for head and neck surgeons. The relative reliability of preoperative orthopantomography, (OPG) bone scanning, computed tomography (CT) and magnetic resonance imaging (MRI), and of peroperative periosteal stripping and direct inspection in clinical assessment for mandibular surgery, is discussed. The histological patterns of tumor invasion and the most common routes of tumor entry in the mandible are described and the influence of variables such as prior radiotherapy and an edentulous vs a dentate state in relation to perineural invasion are also discussed. Finally, a comparison is drawn between the reported outcome of marginal vs segmental resection procedures and a decision-making algorithm is proposed. In selected cases, marginal mandibulectomy can ensure satisfactory tumor control, with a favorable effect on the morbidity associated with mandibular surgery. PMID- 11039867 TI - Nasal airflow in health and disease. AB - This review examines our present understanding of the physiology, pathophysiology and pharmacology of nasal airflow. The main aim of the review is to discuss the basic scientific and clinical knowledge that is essential for a proper understanding of the usefulness of measurements of nasal airflow in the clinical practice of rhinology. The review concludes with a discussion of the measurement of nasal airflow to assess the efficacy of surgery in the treatment of nasal obstruction. Areas covered by the review include: influence of nasal blood vessels on nasal airflow; nasal valve and control of nasal airflow; autonomic control of nasal airflow; normal nasal airflow; nasal cycle; central control of nasal airflow; effect of changes in posture on nasal airflow; effect of exercise on nasal airflow; effect of hyperventilation and rebreathing on nasal airflow; nasal airflow in animals; cerebral effects of nasal airflow; sensation of nasal airflow; sympathomimetics and sympatholytics; histamine and antihistamines; bradykinin; and corticosteroids. PMID- 11039868 TI - Topical aminoglycoside ototoxicity: attempting to protect the cochlea. AB - Cochlear damage following topical application of aminoglycoside antibiotics to the round window membrane is a recognized phenomenon in both animal experiments and clinical reports. The authors have recently reported the ability of the free radical scavenging agent, alpha lipoic acid, to protect against the cochleo-toxic side effects of systemically administered aminoglycoside antibiotics. This study attempts to determine if the protective effect of this free radical scavenging agent is also seen following topical aminoglycoside application. Animals were implanted with osmotic pumps which delivered 2.5 microl/h solution of either neomycin 5% or neomycin plus alpha lipoic acid (50 mg/ml). Control animals received normal saline solution. Drug solutions were presented directly to the round window membrane over a 7-day period. Auditory sensitivity was monitored using compound action potentials (CAPs) of the auditory nerve recorded through an implanted chronic electrode terminating at the round window. Sixteen animals were entered into the study and randomized to one of the above groups. All animals receiving neomycin solution, with or without alpha lipoic acid, maintained normal thresholds for the first 3 days of the treatment period. Animals receiving neomycin solution alone experienced profound and rapid deterioration in auditory sensitivity, which was maximal by day 6. Animals receiving neomycin plus alpha lipoic acid also experienced significant cochlear damage; however, the rate of deterioration was slower than that seen in the group receiving neomycin alone. All control animals receiving saline maintained good hearing thresholds throughout the treatment period. PMID- 11039869 TI - Changes in 2f1 - f2 acoustic distortion products in humans during quinine-induced cochlear dysfunction. AB - Quinine is a suitable model substance for the study of otoacoustic emissions (OAEs) as it reversibly affects the outer hair cells, thus reducing sensitivity, frequency-selectivity and various forms of OAEs. The aim of this experiment was to study quinine-induced changes in the input/output (I/O) function of 2f1 - f2 distortion product OAE (DPOAE; f2/f1 = 1.22; 750-6,000 Hz). Six volunteers with normal hearing (26-39 years old) were intravenously infused to achieve pseudostable quinine plasma concentrations (approximately12 microM) inducing an average pure-tone threshold (PTT; 750-6,000 Hz) shift of 18 dB (5-30 dB) (frequency-independent and reversible). The mean quinine-induced DPOAE shift increased continuously with decreasing equal-level primary tones, e.g. from 1.0 dB at 70 dB sound pressure level (SPL) (n = 42) to 10.5 dB (n = 22) at 40 dB SPL (pooled data, no frequency dependence). According to recruitment, the mean slope of the DPOAE I/O function (at 30-60 dB SPL) increased from 0.86 to 1.35 dB/dB. The lack of correlation between shifts in DPOAE and PTT is in stark contrast to the excellent correlation reported between shifts in transient evoked OAE detection threshold and its corresponding psychoacoustic threshold. The highly vulnerable spontaneous OAEs, in combination with the less vulnerable DPOAEs, fit into a recently proposed taxonomic classification for OAEs. PMID- 11039870 TI - Expression of vascular cell adhesion molecule-1, alpha4-integrin and L-selectin during inner ear immunity reaction. AB - The expression of vascular cell adhesion molecule-1 (VCAM-1), alpha4-integrin and L-selectin was dynamically observed using an immunohistochemical approach during labyrinthitis induced by inoculation of keyhole limpet hemocyanin into the scala tympani of animals that had been systemically sensitized to it. At the same time, we used ELISA to monitor interleukin-1(IL-1) in the peripheral blood of these animals. The expression of alpha4-integrin and L-selectin on the surface of leukocytes in peripheral blood was examined by flow cytometry. VCAM-1 was found on the endothelial surface of the spiral modiolar vein (SMV) and its collecting venules (CV) at 24 h post-challenge; the VCAM-1 level reached a maximum on day 2, which was maintained until 1 week post-challenge, and then declined gradually. Alpha4-integrin was found on the surface of leukocytes, mainly monocytes and lymphocytes, that had infiltrated the SMV, CV, cochlear nerve and perilymph by 24 h post-challenge; the alpha4-integrin level reached a maximum on day 2 and then decreased rapidly. At 1 week post-challenge, no significant expression of alpha4 integrin was seen. Expression of alpha4-integrin and L-selectin was also observed on leukocytes in peripheral blood. No L-selectin was observed on the surface of leukocytes during inner ear inflammation. The expression of L-selectin on the surface of leukocytes in the peripheral blood appeared to be downregulated during inflammation. The concentrations of IL-1 in the peripheral blood increased commensurately during inner ear inflammation. These results further elucidate the role of adhesion molecules during inner ear immune responses. The increased expression of VCAM-1 on SMV and CV was correlated with the concentration of IL-1 in peripheral blood during inner ear inflammation. PMID- 11039871 TI - Magnetic resonance imaging of the temporal bone in patients with Meniere's disease. AB - Meniere's disease (MD) is still controversial in several aspects. The vestibular aqueduct, the osseous channel that carries the endolymphatic duct and sac, has previously been studied by tomography and computed tomography, with shortening and narrowing of this structure observed. These findings are apparently correlated to the development of the endolymphatic hydrops present in MD and related to its episodic symptoms. In studying the endolymphatic duct, the key structure in the pathology of this disease, magnetic resonance imaging (MRI) studies of the temporal bone were performed in 12 patients with unilateral MD and in 9 bilateral cases; the results were compared with images obtained from 30 normal ears. The endolymphatic duct appeared to be statistically less visible in MD patients, with no difference between symptomatic and asymptomatic ears in the presence of unilateral disease. No relationship was found between visualization of the endolymphatic duct and time of evolution or response to clinical treatment in these cases. The distance from the posterior semicircular canal to the posterior temporal border was found to be bilaterally reduced in MD. The authors conclude that although the demonstration of endolymphatic hydrops "in vivo" is not yet possible by MRI, some features can be observed that can support a clinical hypothesis of MD. PMID- 11039872 TI - Long-term evolution of subjective visual vertical after vestibular neurectomy and labyrinthectomy. AB - Subjective visual vertical (SVV) tilt, observed after vestibular neurectomy and labyrinthectomy, is considered to be due to the deafferentation of the otolithic organs. The aim of this study was to determine the long-term evolution of the SVV up to 4 years after surgery. Between 1993 and 1998 the SVV was measured in 35 patients (18 men, 17 women) aged from 21 to 71 years (mean 36 years). Vestibular neurectomy was performed in 30 patients and labyrinthectomy in 6. SVV was measured with a binocular test (vertical frame) and a monocular test (Maddox rod). Immediately after operation, the SVV showed a 10-30 degrees tilt toward the operated ear with the vertical frame (normal 0 +/- 2 degrees) and a 5-22 degrees tilt with the Maddox rod (normal 0 +/- 4 degrees). After labyrinthectomy, SVV returned to normal values after 1 year in all patients. SVV also returned to normal within 1 year after vestibular neurectomy in 20 patients; in the other 10 patients SVV was still slightly tilted 1-4 years after neurectomy: > 2 degrees with the vertical frame and > 4 degrees with the Maddox rod, particularly on the eye ipsilateral to the operated ear. Some of these 10 patients also had persisting disequilibrium. PMID- 11039873 TI - Medial (intra-cisternal) acoustic neuromas. AB - The clinical characteristics of "medial" or "intra-cisternal" acoustic neuroma (AN) treated in our institute were reviewed. Among 466 patients with ANs in our series during the last 20 years, 6 patients (1.3%) were considered to fill the criteria of medial AN definition. Compared with those with non-medial ANs, the patients with medial ANs show a tendency to have cerebellar and/or cranial nerve dysfunction (especially trigeminal and/or facial nerves) in addition to hearing loss at the time of initial presentation. On magnetic resonance imaging, medial AN is visualized as a multi-cystic mass lesion in the cerebello-pontine cistern without extension into the internal auditory canal in most cases. Although total removal of tumor was achieved in all cases, the results of preservation of facial nerve function were not satisfactory. Medial AN can be considered as a clinical, but not pathological, subtype in terms of the functional outcomes of the facial nerve and hearing. PMID- 11039874 TI - Acoustic neuroma: deterioration of speech discrimination related to thresholds in pure-tone audiometry. AB - Pure-tone and speech audiometry were performed in 231 patients with a unilateral acoustic neuroma. Tumor sizes were obtained through imaging. Audiometric parameters, such as the mean pure-tone thresholds, the maximum discrimination, the slope of the speech audiogram, the roll-over index and the difference between the speech reception threshold and the Fletcher index, were studied and compared with data in the literature. Results showed that patients with an acoustic neuroma have worse speech discrimination than can be expected from the pure-tone audiogram. However, the results presented here indicate that hearing impairments nowadays are not as severe as those described in earlier studies. More patients with a unilateral acoustic neuroma are detected, including even those with a minor hearing impairment. The roll-over index is not characteristic for patients with an acoustic neuroma. PMID- 11039875 TI - Age-related hearing impairment and B vitamin status. AB - The aim of this study was to examine, in elderly subjects, a possible association between age-related hearing impairment and vitamin B12 or folic acid status. Ninety-one consecutive subjects with pure age-related hearing impairment, 35 males and 56 females, with a median age of 78 years, range 67-88 years, were included in the investigation. All subjects underwent a thorough evaluation, including pure-tone, speech and impedance audiometry. Blood samples were drawn for determination of B12, folic acid and homocysteine and analysed by routine laboratory measurements. No significant differences in the blood parameters as a function of gender could be demonstrated and no correlations were found between B12 or folic acid and hearing levels averaged across the range 0.5-4 kHz. A weak correlation between hearing levels and homocysteine (r = 0.03; correlation coefficient 0.004) was found; however, a comparison of the hearing levels between those with increased and normal homocysteine failed to show any significant differences. In addition, no association between B12 and the speech recognition score could be found. This investigation therefore fails to demonstrate any association between hearing level and vitamin B12 or folic acid in elderly subjects. PMID- 11039876 TI - Neural representation of sound duration in the inferior colliculus of the mouse. AB - In this study we examined the neuronal responses of single units to different sound durations in the inferior colliculus (IC) of the mouse. One hundred and one recorded units were classified into onset (58%), sustained (9%) on-sustained (22%), pauser (9%) and chopper (2%) response patterns. Thirty-four percent of the recorded units showing stronger responses to long stimulus durations were defined as long-duration-selective neurons. Twenty-five percent of the units preferred a narrow range of sound durations and were classified as band-pass neurons. Ten percent of the units responded preferentially to short stimulus durations and thus displayed short-duration selectivity. Twelve percent of the units that responded with nearly constant spike counts to stimuli of varying duration were classified as all-pass neurons. In contrast to the result of no short-duration selective neurons found in chinchilla IC, we observed that some of the onset units in the IC of the mouse displayed a short duration preference. The best duration range of the duration-selective neurons in the present study corresponds to the duration range of mouse calls. We suggest that an inhibitory mechanism contributes to the duration selectivity observed in the present study. PMID- 11039877 TI - The role of thymidine phosphorylase in the pathogenesis of allergic rhinitis. AB - The activity and distribution of thymidine phosphorylase (TP) in the nasal mucosa of patients with nasal allergy was examined and compared with those in healthy subjects. TP activity was analyzed by spectrophotometry and expression was examined by immunoblotting and immunohistochemical staining using a monoclonal antibody specific to TP. The expression level of TP detected by immunoblotting showed a correlation with the activity detected by spectrophotometry. In nasal mucosa obtained from patients with nasal allergy, the level of TP was significantly higher than that from normal subjects. Eosinophils, basal cells in mucosal epithelium and fibroblasts in nasal mucosa obtained from patients with nasal allergy were stained with anti-TP monoclonal antibody. Strong staining of eosinophils present in nasal discharge was observed. The present results indicate that an increased number of TP-expressing cells, especially eosinophils in nasal mucosa, might be associated with the pathogenesis of nasal allergy. PMID- 11039878 TI - Production of interferon-gamma by tonsillar mononuclear cells in IgA nephropathy patients. AB - Much evidence, both in vivo and in vitro, suggests that patients with IgA nephropathy (IgAN) have enhanced IgA production. We hypothesized that Haemophilus parainfluenzae (HP) in the tonsil plays an important role in the IgA production of IgAN patients. In this study, we focused on interferon-gamma (IFN-gamma) and IgA production by tonsillar mononuclear cells (TMC) in patients with IgAN. Tonsillectomies were performed in patients with IgAN and chronic tonsillitis (CT). The induction of IFN-gamma and IgA in vitro by TMC from IgAN patients was compared with that from CT patients. In addition, we investigated whether stimulation with the outer membranes of HP (OMHP) enhanced IFN-gamma and IgA induction by TMC from patients with IgAN. Spontaneous IFN-gamma production and spontaneous total IgA production by TMC were higher in IgAN patients than in those patients with CT (p < 0.05). IFN-gamma induction by OMHP stimulation was also higher in IgAN patients than in CT patients. The stimulation of OMHP enhanced HP-specific IgA in IgAN but had no influence on the production of IFN gamma in patients with either IgAN or CT compared with spontaneous IFN-gamma production. IFN-gamma production was positively correlated with total IgA values in both IgAN and CT patients, but not with HP-specific IgA. These results suggest that increased IFN-gamma production in patients with IgAN is not associated with HP infection but may play a role in the pathogenesis of IgAN. PMID- 11039879 TI - Laryngeal findings in patients with contact granuloma: a long-term follow-up study. AB - In order to evaluate the long-term outcome of contact granuloma a follow-up study of 59 patients was conducted. Primary treatment consisted mainly of voice therapy or of voice therapy in combination with surgery. A careful history was taken and the patients were examined using videolaryngoscopy. The average follow-up time was 12 years (range 5-20 years). In total, 10 out of 59 patients (17%) had a granuloma at the follow-up examination, 6 of them presenting without subjective symptoms. Another 28 patients had various signs of inflammation of the posterior glottis. The history taken at follow-up revealed that eight patients had had a previous temporary recurrence during the follow-up time. The recurrence frequency was the same for patients who received voice therapy after postoperative recurrence and patients who were treated with voice therapy only. About 80% of the patients had no symptoms at follow-up. The findings of the present study indicate that contact granuloma is one of several manifestations of chronic posterior laryngitis and that it may occur without symptoms. PMID- 11039880 TI - Characterization of AMC-HN-9, a cell line established from an undifferentiated carcinoma of the parotid gland: expression of alpha6beta4 with the absence of BP180 and 230. AB - We recently reported the development of a cell line, AMC-HN-9, established from an undifferentiated carcinoma (UDC) of the parotid gland. AMC-HN-9 consists mostly of spindle-shaped cells, has poor in vitro adhesiveness and an in vitro appearance that is different from that of other epithelial cell lines. To test the hypothesis that structural or functional abnormalities of the hemidesmosomes might contribute to the morphological appearance and biology of UDCs, we studied the expression of hemidesmosomal proteins in AMC-HN-9. Flow cytometry, indirect immunofluorescence, immunoprecipitation, reverse transcriptase-polymerase chain reaction, and cytogenetic analysis were used. AMC-HN-9 cells express the alpha6 and beta4 integrin subunits at nearly the same intensity as head and neck squamous cell carcinoma cell lines. However, AMC-HN-9 does not express BP180 and BP230, although there is no gross deletion of the loci of the BP180 and BP230 genes, suggesting that a more subtle mechanism has silenced these genes. In conclusion, the failure to express certain hemidesmosomal proteins is a likely explanation for the functional and morphologic characteristics of UDC cells both in vivo and in vitro. PMID- 11039881 TI - Edematous swelling of the facial nerve in Bell's palsy. AB - Surgical decompression of the intratemporal facial nerve from the geniculate ganglion to the stylomastoid foramen was carried out in 91 patients with Bell's palsy. All of the patients had denervation exceeding 95%, and a suprastapedial lesion. Edematous swelling of the nerve was assessed using the following three grades: + +, nerve swells beyond the bony facial canal; +, nerve swells beyond the nerve sheath, but within the bony canal, and -, no notable swelling observed. Varying degrees of swelling of the nerve were noted in all of the patients from onset to the end of the ninth week. The incidence of + + swelling was highest within 3 weeks of onset and then declined. + + swelling was most often noted in the vicinity of the geniculate ganglion, and was thought to be a manifestation of inflammation due to herpes simplex virus infection. There was a clear time dependency of the swelling in the horizontal and pyramidal segments, but not in the mastoid segment. After the ninth week, the incidence of swelling decreased sharply and no swelling of the nerve was observed in about one-third of the patients. Considering the etiology and the analysis of edematous swelling, we propose that the course of Bell's palsy be differentiated into an acute phase (the first 3 weeks after onset), a subacute phase (from the fourth to ninth weeks) and a chronic phase (after the tenth week). PMID- 11039882 TI - New technology to control symptoms in Meniere's disease. PMID- 11039883 TI - Overview: soft tissue augmentation. AB - No single filler substance meets all of a clinician's expectations. There are, however, many substances currently available that are useful in the soft tissue armamentarium. This article reviews the many filler substances; their indications, shortcomings, complications, and approval status by the Federal Food and Drug Administration. PMID- 11039884 TI - Response to tissue injury. AB - Cutaneous injury, whether by laser, chemical, or scalpel, results in scar formation. The normal response to such an insult occurs in the middle of a continuum of wound repair processes. On one end of the continuum are the overhealed responses (i.e., keloids). On the opposite end are tissue regeneration and scarless healing as seen in fetal wounds. This article reviews the molecular biology and mechanisms leading to these various clinical phenotypes and discusses future potential modalities that may replicate the scarless wound healing mechanism in adults. PMID- 11039885 TI - Skin care and the topical treatment of aging skin. AB - The desire to maintain or regain a youthful appearance is the main motivation of patients who present themselves to the aesthetic plastic surgeon's office. It has become imperative for the plastic surgeon to know and understand the causes of aging and skin damage and to provide ancillary nonsurgical treatments through which their patients can achieve rejuvenation. PMID- 11039886 TI - Autologen. AB - This article gives an overview of autologous injectable collagen and reviews the theoretical complications and issues of using an autologous filler material. The benefits of autologous injectable collagen include elimination of local tissue reaction and rejection. The product, however, has not yet been proven to have significant longevity beyond that of its competitors. PMID- 11039887 TI - Long-term outcome of autologous fat transplantation in aesthetic facial recontouring: sixteen years of experience with 1936 cases. AB - Fat grafts have become an important and necessary procedure in the field of plastic surgery. The author presents experiences with two types of patients: older people with thin facial soft tissues, in whom the combination of lifting and lipoinjection gives good results; and patients with progressive hemifacial atrophy caused by Parry-Romberg's syndrome in whom normal or almost normal facial contour can be achieved. The longterm survival of fat grafts is presented. This procedure has been used by the author for the past 16 years. PMID- 11039888 TI - Fat transfer: current use in practice. AB - The author has used fat transfer, including pearl fat grafts and fat injections, for almost 18 years in practice. Techniques for pearl fat grafting and fat injections are described. Pearls are limited to eyelids and small depressions. Fat injections can be used to augment various facial areas, including chin, cheekbones, nasolabial folds, lips, labiomandibular folds, glabella, forehead, and nose. PMID- 11039889 TI - Porous hydroxyapatite granules for alloplastic enhancement of the facial region. AB - Hydroxyapatite is a biocompatible alloplast with the same chemical composition as bone. It is readily incorporated into host bone, does not undergo appreciable resorption, does not incite a clinically significant foreign body reaction, and resists infection. This article describes forms of hydroxyapatite, procedures for use, and clinical examples. PMID- 11039890 TI - Microdermabrasion in clinical practice. AB - Microdermabrasion is the general term applied to the technique of abrading the skin with a high-pressure flow of crystals. Patient selection, equipment, and technique for face, hands, and chest are described. Results of a survey on the efficacy of the treatment are provided. PMID- 11039891 TI - Use of expanded polytetrafluoroethylene in aesthetic surgery of the face. AB - Aesthetic surgery procedures and materials available to improve the appearance of various subunits of the face have been evolving for the past several decades. Initially, techniques such as direct excision or placement of autogenous tissue grafts (e.g., fat, fascia, tendon, and dermal fat grafts) were used. Additional options became available with the development of silicone, injectable collagen, and ePTFE. These options for facial augmentation can be done rapidly in an office setting and as an outpatient. An important aspect of treatment is discussing clearly all pertinent options with the patient, and recommending the appropriate option based on the patient's anatomic findings and personal needs. For example, patients desiring lip augmentation but unsure about the result may be best treated with collagen injection, which gives temporary results and lets the patient consider a long-term result provided by ePTFE or autogenous grafts. Use of ePTFE in facial aesthetic surgery gradually has increased over the past two decades. Its use in augmentation of lips, nasolabial creases, and nose is becoming an important treatment modality for aesthetic surgeons. PMID- 11039892 TI - Liquid injectable silicone: techniques for soft tissue augmentation. AB - This article includes a description of the uses and indications of liquid injectable silicone and a discussion of the results obtained. Patient selection, indications, mechanism of action, potential complications, and adjunct procedures are also addressed. PMID- 11039893 TI - Autologous cultured fibroblasts as cellular therapy in plastic surgery. AB - Autologous cultured fibroblasts serve as injectable protein repair systems for correction of acne scars, rhytids, and other facial scars. The system uses the patient's own cultured fibroblasts to correct contour deformities over time. PMID- 11039894 TI - Facial scars. AB - The management of facial scars is a challenging problem, even for the experienced physician. Careful analysis of the scar's visibility and unpleasantness to the patient is necessary before determining a treatment plan. Multiple modalities are often needed to achieve optimal results. PMID- 11039895 TI - Biologic therapy. AB - Biologic therapy has evolved as a fourth treatment option for patients with melanoma and other malignancies. Interrelated advances in molecular diagnostics and gene therapy techniques will further accelerate therapeutic advances in this new arena. A variety of biologic therapeutic options available to the patient with high risk melanoma are reviewed. PMID- 11039896 TI - Human gelsolin prevents apoptosis by inhibiting apoptotic mitochondrial changes via closing VDAC. AB - Gelsolin is a Ca2+-dependent actin-regulatory protein that modulates actin assembly and disassembly, and is believed to regulate cell motility through modulation of the actin network. Gelsolin was also recently suggested to be involved in the regulation of apoptosis: human gelsolin (hGsn) has anti-apoptotic activity, whereas mouse gelsolin (mGsn) exerts either proapoptotic or anti apoptotic activity depending on different cell types. Here, we studied the basis of anti-apoptotic activity of hGsn. We showed that both endogenous and overexpressed hGsn has anti-apoptotic activity, that depends on its C-terminal half. We also found that hGsn and its C-terminal half but not mGsn could prevent apoptotic mitochondrial changes such as Apsi loss and cytochrome c release in isolated mitochondria to a similar extent as Bcl-xL, indicating that hGsn targets the mitochondria to prevent apoptosis via its C-terminal half. In the same way as anti-apoptotic Bcl-xL, which we recently found to prevent apoptotic mitochondrial changes by binding and closing the voltage-dependent anion channel (VDAC), hGsn and its C-terminal half inhibited the activity of VDAC on liposomes through direct binding in a Ca2+-dependent manner. These results suggest that hGsn inhibits apoptosis by blocking mitochondrial VDAC activity. PMID- 11039897 TI - Oncogenic transformation by the FOX protein Qin requires DNA binding. AB - Some functions of the Qin oncoprotein are not dependent on DNA binding. In order to test the requirement for DNA binding in Qin-induced oncogenic transformation, site directed mutations were introduced in the winged helix (WH) DNA binding domain of the Qin protein. In cellular Qin (c-Qin), the glycine at position 233 was either deleted or substituted with the amino acids aspartic acid, alanine, glutamic acid, asparagine, proline or lysine. The same position carries aspartic acid in the viral Qin protein (v-Qin). The adjacent residues, threonine 232 and lysine 234, were separately mutated to proline. Several additional amino acid substitutions believed to be involved in DNA contacts were introduced at the following c-Qin positions: asparagine 189, histidine 193, serine 196 or arginine 236. Most of the substitutions reduced DNA binding of Qin, one mutation, H193A, completely abolished DNA binding, and another mutation, T232P, increased DNA binding affinity. Mutant H193A failed to transform chicken embryo fibroblasts (CEF), all other mutants, even those showing minimal DNA binding, retained oncogenicity for CEF. The efficiencies of focus formation induced by these mutant proteins in cell culture were not significantly different from that of wild type. However, the rate of focus development and the size of foci induced by the Qin mutants were greater with strong DNA binders than with weak DNA binders. Transdominant negative constructs consisting of the winged helix domain of cQin or v-Qin interfered with focus formation induced by full length Qin proteins. These results suggest that DNA binding is a prerequisite for transformation by Qin, and strong DNA binding is related to accelerated transformation in CEF. PMID- 11039898 TI - p27kip1-independent cell cycle regulation by MYC. AB - MYC transcription factors are potent stimulators of cell proliferation. It has been suggested that the CDK-inhibitor p27kip1 is a critical G1 phase cell cycle target of c-MYC. We show here that mouse embryo fibroblasts deficient for both p27kip1 and the related p21cip1 are still responsive to stimulation by c-MYC and can be arrested in G1 by a dominant negative mutant of c-MYC. This growth arrest can be overruled by ectopic expression of E2F or adenovirus E1A, but not by a mutant of E1A defective for binding to retinoblastoma family proteins. We show that fibroblasts with a genetic disruption of all three retinoblastoma family members (pRb, p107 and p130) are unresponsive to a dominant negative c-MYC mutant. These data indicate that p27kip1 is not the only rate limiting cell cycle target of c-MYC and suggest that regulation of E2F is also essential for c-MYC's mitogenic activity. PMID- 11039899 TI - Complementation of Myc-dependent cell proliferation by cDNA expression library screening. AB - The targeted knockout of the c-myc gene from rat fibroblasts leads to a stable defect in cell proliferation. We used complex cDNA libraries expressed from retroviral vectors and an efficient sorting procedure to rapidly select for cDNAs that can restore the growth rate of c-myc deficient cells. All of the biologically active cDNAs contained either c-myc or N-myc, suggesting that no other cellular genes can effectively bypass the requirement for c-myc in fibroblast proliferation. This approach provides a powerful screening method for cell cycle changes in genetically defined systems. PMID- 11039900 TI - Adhesion induced expression of the serine/threonine kinase Fnk in human macrophages. AB - Members of the polo subfamily of protein kinases play crucial roles in cell proliferation. To study the function of this family in more detail, we isolated the cDNA of human Fnk (FGF-inducible kinase) which codes for a serine/threonine kinase of 646 aa. Despite the homology to the proliferation-associated polo-like kinase (Plk), tissue distribution of Fnk transcripts and expression kinetics differed clearly. In contrast to Plk no correlation between cell proliferation and Fnk gene expression was found. Instead high levels of Fnk mRNA were detectable in blood cells undergoing adhesion. The transition of monocytes from peripheral blood to matrix bound macrophages was accompanied by increasing levels of Fnk with time in culture. Neither treatment of monocytes with inducers of differentiation nor withdrawal of serum did influence Fnk mRNA levels significantly, suggesting that cell attachment triggers the onset of Fnk gene transcription. The idea that Fnk is part of the signalling network controlling cellular adhesion was supported by the analysis of the cytoplasmic distribution of the Fnk protein and the influence of its overexpression on the cellular architecture. Fnk as fusion protein with GFP localized at the cellular membrane in COS cells. Dysregulated Fnk gene expression disrupted the cellular f-actin network and induced a spherical morphology. Furthermore, Fnk binds to the Ca2+/integrin-binding protein Cib in two-hybrid-analyses and co immunoprecipitation in assays. Moreover, both proteins were shown to co-localize in mammalian cells. The homology of Cib with calmodulin and with calcineurin B suggests that Cib might be a regulatory subunit of polo-like kinases. PMID- 11039901 TI - Pituitary neoplasia induced by expression of human neurotropic polyomavirus, JCV, early genome in transgenic mice. AB - In recent years, there has been mounting evidence pointing to the association of polyomaviruses with a wide range of human cancers. The human neurotropic polyomavirus, JCV, infecting greater than 75% of the human population produces a regulatory protein named T-antigen which is expressed at the early phase of viral lytic infection and plays a critical role in completion of the viral life cycle. Furthermore, this protein has the ability to transform neural cells in vitro and its expression has been detected in several human neural-origin tumors. To further investigate the oncogenic potential of the JCV early protein in vivo, transgenic mice expressing JCV T-antigen under the control of its own promoter were generated. Nearly 50% of the animals developed large, solid masses within the base of the skull by 1 year of age. Evaluation of the location as well as histological and immunohistochemical data suggest that the tumors arise from the pituitary gland. As T-antigen is known to interact with several cell cycle regulators, the neoplasms were analysed for the presence of the tumor suppressor protein, p53. Immunoprecipitation/Western blot analysis demonstrated overexpression of wild-type, but not mutant p53 within tumor tissue. In addition, co-immunoprecipitation established an interaction between p53 and T-antigen and overexpression of p53 downstream target protein, p21/WAF1. This report describes the analysis of inheritable pituitary adenomas induced by expression of the human polyomavirus, JCV T-antigen in transgenic mice where T-antigen disrupts the p53 pathway by binding to and sequestering wild-type p53. This animal model may serve as a useful tool to further evaluate mechanisms of tumorigenesis by JCV T antigen. PMID- 11039902 TI - Restoration of positioning control following Disabled-2 expression in ovarian and breast tumor cells. AB - The physical interaction of epithelial cells with the basement membrane ensures correct positioning and acts as a survival factor for epithelial cells. Cells that detach from the basement membrane often undergo apoptosis; however, in carcinomas, this positional control is absent, permitting disorganized cell proliferation. In the majority of breast and ovarian carcinomas (85-90%), the expression of a candidate tumor suppressor, Disabled-2 (Dab2), is frequently lost. The Dab2-negative tumor cells are no longer in contact with an intact basement membrane, as indicated by the absence of collagen IV (in about 90% of cases). However, in the subset (10-15%) of ovarian tumors in which Dab2 expression is positive, the presence of a basement membrane-like structure around tumor cells was observed. Recombinant adenovirus-mediated expression of Dab2 was used in Dab2-negative ovarian and breast cancer cells, and re-expression of Dab2 was found to lead to cell death or growth arrest. Dab2 expression suppressed MAPK activation and c-fos expression. Plating the infected cells on a basement membrane matrigel rescued the cells from death and growth arrest. Thus, Dab2 exhibits a negative activity for cell growth and survival, which can be countered by attachment of the cells to basement membrane matrix. We conclude that Dab2 functions in cell positioning control and mediates the exigency for basement membrane attachment of epithelial cells. Loss of Dab2 may contribute to the basement membrane-independent, disorganized proliferation of tumor cells in ovarian and breast carcinomas. PMID- 11039903 TI - Differential regulation of HSP27 oligomerization in tumor cells grown in vitro and in vivo. AB - HSP27 form oligomeric structures up to 800 Kda. In cultured cells, the equilibrium between small and large oligomers shifted towards smaller oligomers when phosphorylated on serine residues. To further explore HSP27 structural organization and its repercussion in HSP27 antiapoptotic and tumorigenic properties, we transfected colon cancer REG cells with wild type HSP27 and two mutants in which the phosphorylatable serine residues have been replaced by alanine (to mimic the non phosphorylated protein) or aspartate (to mimic the phosphorylated protein). In growing cells, wild type and alanine mutant formed small and large oligomers and demonstrated antiapoptotic activity while aspartate mutant only formed small multimers and had no antiapoptotic activity. In a cell free system, only large oligomeric structures interfered with cytochrome c induced caspase activation, thereby inhibiting apoptosis. The inability of the aspartate mutant to form large oligomers and to protect tumor cells from apoptosis was overcome by growing the cells in vivo, either in syngeneic animals or nude mice. These observations were reproduced by culturing the cells at confluence in vitro. In conclusion (1) large oligomers are the structural organization of HSP27 required for its antiapoptotic activity and (2) cell-cell contacts induce the formation of large oligomers, whatever the status of phosphorylatable serines, thereby increasing cell tumorigenicity. PMID- 11039904 TI - C-erbB-2/ HER-2 upregulates fascin, an actin-bundling protein associated with cell motility, in human breast cancer cell lines. AB - The over-expression of c-erbB-2/ HER-2, a receptor tyrosine kinase, correlates with poor prognosis in patients with breast and ovarian cancer. In the human breast cancer cell line, MDA-MB-435, c-erbB-2 over-expression results in increased chemoinvasion and higher metastatic properties in nude mice. However, the mechanisms by which c-erbB-2 increases the malignant potential of cells remains unclear. We have determined that over-expression of c-erbB-2 in MDA-MB 435 cells, and in some additional breast cancer cell lines, is associated with graphic increases in mRNA and protein levels of the actin bundling protein fascin. Heightened fascin expression has been observed in other systems to result in greatly increased cell motility, and indeed, our work employing semi-automated time-lapse microscopy demonstrates that MDA-MB-435 cells over-expressing c-erbB-2 exhibit significantly heightened cellular dynamics and locomotion, while visualization of bundled microfilaments within fixed cells revealed enhanced formation of dendritic-like processes, microspikes and other dynamic actin based structures. To address the means by which c-erbB-2 over-expression might result in elevated fascin levels, we identified multiple perfect match TCF and NF-kappaB consensus sites in fascin's promoter and first intron, which appeared consistent with the greater endogenous transcriptional activities of TCF and NF-kappaB in c erbB-2 over-expressing MDA-MB-435 cells. While such transcriptional modulation may occur in the context of the intact gene/chromatin, subsequent tests using reporter constructs did not support involvement of these signaling pathways. In conclusion, highly increased fascin levels were observed in MDA-MB-435 over expressing c-erbB-2, likely contributing to these cells' altered actin dynamics, and increased cell motility and malignancy. Studies in progress aim to discern the means by which c-erbB-2 over-expression leads to transcriptional activation of the fascin gene. PMID- 11039905 TI - Directed mutation of the basic domain of v-Jun alters DNA binding specificity and abolishes its oncogenic activity in chicken embryo fibroblasts. AB - Overexpression of v-Jun in chicken embryo fibroblasts (CEF) leads to oncogenic transformation phenotypically characterized by anchorage independent growth and release from contact inhibition (focus formation). The mechanisms involved in this oncogenic conversion however, are not yet clear. Because Jun is a transcription factor, it has been assumed that oncogenic transformation results directly from deregulated AP-1 target gene expression. However, a number of experimental observations in avian cell culture models fail to correlate oncogenesis with AP-1 activity suggesting that transformation induced by v-Jun may occur through an indirect mechanism. To test this possibility, we introduced point mutations into the basic DNA binding domain of v-Jun and created mutants that exhibit altered binding specificity. When expressed in CEF, these mutants fail to deregulate three known v-Jun target genes (JTAP-1, apolipoprotein A1, c Jun) thus demonstrating in vivo specificity changes. Each of the binding specificity mutants was also tested for its ability to induce oncogenic transformation. Interestingly, expression of these mutants in CEF results in a phenotype indistinguishable from the vector control with respect to growth rate, focus formation and the ability to form colonies in soft agar. These results are consistent with a model requiring direct AP-1 target deregulation as a prerequisite of v-Jun induced cell transformation. With this in mind, we generated a series of additional mutants that retain the ability to bind AP-1 sequence elements, but vary in their oncogenic potential. We demonstrate the use of these mutants to screen v-Jun induced gene targets for a functional role in cell transformation. PMID- 11039906 TI - B-Myc is preferentially expressed in hormonally-controlled tissues and inhibits cellular proliferation. AB - The myc family of genes plays an important role in several cellular processes including proliferation, apoptosis, differentiation, and transformation. B-myc, a relatively new and largely unstudied member of the myc family, encodes a protein that is highly homologous to the N-terminal transcriptional regulatory domain of c-Myc. Here, we show that high level B-myc expression is restricted to specific mouse tissues, primarily hormonally-controlled tissues, with the highest level of expression in the epididymis. We also report the identification of the endogenous B-Myc protein from mouse tissues. Like other Myc family proteins, B-Myc is a short-lived nuclear protein which is phosphorylated on residues Ser-60 and Ser 68. Rapid proteolysis of B-Myc occurs via the ubiquitin-proteasome pathway. Finally, we found that overexpression of B-Myc significantly slows the growth of Rat la fibroblasts and COS cells suggesting B-Myc functions as an inhibitor of cellular proliferation. PMID- 11039907 TI - Role of the cAMP and MAPK pathways in the transformation of mouse 3T3 fibroblasts by a TSHR gene constitutively activated by point mutation. AB - Constitutive activating mutations of the TSHR gene, have been detected in about 30 per cent of hyperfunctioning human thyroid adenomas and in a minority of differentiated thyroid carcinomas. The mutations activating the TSHR gene(s) in the thyroid carcinomas, were located at the codon 623 changing an Ala to a Ser (GCC-->TCC) or in codon 632 changing a Thr to Ala or Ile (ACC-->GCC or ACC- >ATC). In order to study if the constitutively activated TSHR gene(s) has played a role in the determination of the malignant phenotype presented by these tumors, we investigated: (1) the transforming capacity after transfection of mouse 3T3 cells, of a TSHR cDNA activated by an Ala-->Ser mutation in codon 623 or an Thr- >Ile mutation in codon 632 and (2) the pathway(s) eventually responsible(s) for the malignant phenotype of the cells transformed by these constitutively activated TSHR cDNAs. Our results show that (1) the TSHR(M623) or (M632) cDNAs give rise to 3T3 clones presenting a fully neoplastic phenotype (growth in agar and nude mouse tumorigenesis); this phenotype was weaker in the cells transformed by the 632 cDNA; (2) suggest that the fully transformed phenotype of our 3T3 cells, may be the consequence of the additive effect of the activation of at least two different pathways: the cAMP pathway through G(alpha)s and the Ras dependent MAPK pathway through G(beta)gamma and PI3K and (3) show that the PI3K isoform playing a key role as an effector in the MAPK pathway activation in our 3T3-transformed cells is PI3Kgamma. Signaling from PI3Kgamma to MAPK appears to require in our murine cellular system a tyrosine kinase (still not characterized), Shc, Grb2, Sos, Ras and Raf. It is proposed that the constitutively activated TSHR genes detected in the thyroid carcinomas, may have played an oncogenic role, participating in their development through these two pathways. PMID- 11039908 TI - The mitotic serine/threonine kinase Aurora2/AIK is regulated by phosphorylation and degradation. AB - Aurora2 is a cell cycle regulated serine/threonine protein kinase which is overexpressed in many tumor cell lines. We demonstrate that Aurora2 is regulated by phosphorylation in a cell cycle dependent manner. This phosphorylation occurs on a conserved residue, Threonine 288, within the activation loop of the catalytic domain of the kinase and results in a significant increase in the enzymatic activity. Threonine 288 resides within a consensus motif for the cAMP dependent kinase and can be phosphorylated by PKA in vitro. The protein phosphatase 1 is shown to dephosphorylate this site in vitro, and in vivo the phosphorylation of T288 is induced by okadaic acid treatment. Furthermore, we show that the Aurora2 kinase is regulated by proteasome dependent degradation and that Aurora2 phosphorylated on T288 may be targeted for degradation during mitosis. Our experiments suggest that phosphorylation of T288 is important for regulation of the Aurora2 kinase both for its activity and its stability. PMID- 11039909 TI - Regulation of proliferation-survival decisions is controlled by FGF1 secretion in retinal pigmented epithelial cells. AB - Fibroblast growth factor 1 (FGF1) induces proliferation and differentiation in a wide variety of cells of mesodermal and neuroectodermal origin. FGF1 has no 'classical' signal sequence to direct its secretion, and there has been considerable debate concerning FGF1 secretion and its role in the biological activities of FGF1. We investigated the effects of FGF1 secretion and the signalling induced by signal peptide (SP)-containing FGFI and SP-less FGF1, on the proliferation and the apoptosis in retinal pigmented epithelial (RPE) cells. Primary RPE cell cultures were transfected with FGF1 (FGF1 cells) and SP-FGF1 (SP FGF1 cells) cDNAs. SP-FGF1 cells secreted large amount of FGF1 and actively proliferated, whereas FGF1 and control cells did not. Secreted FGF1 induced short term activation of both FGFR1 and ERK2, which were required for cell proliferation. In contrast, SP-FGF1 cells stopped secreting FGF1 and died rapidly, if cultured in the absence of serum. Surprisingly, FGF1 cells, but not control cells, secreted FGF1 and were resistant to apoptosis induced by serum depletion. Secreted FGF1 induced long-term activation of FGFR1 and ERK2, which was necessary to induce a constant and high level of Bcl-x production, and to induce cell survival in FGFI cells. Downregulation of ERK2 and Bcl-x increased apoptosis. Thus, the proliferation and survival activities of FGF1 depend on the secretion of FGF1 which is determined by the cell culture conditions. Cell proliferation was SP-dependent, whereas cell survival was not. The signal peptide controls the level and duration, 'whispering or shouting', of ERK2 activation cells which determines FGF1 biological function and may have important implications for anti-degenerative and anti-proliferative treatments. PMID- 11039910 TI - Transcriptional regulation of the major histocompatibility complex (MHC) class I heavy chain, TAP1 and LMP2 genes by the human papillomavirus (HPV) type 6b, 16 and 18 E7 oncoproteins. AB - We have examined the possibility that the E7 proteins of the high-risk human papillomavirus (HPV) type 16 and 18 and the oncogenic adenovirus (Ad) type 12 E1A protein share the ability to down-regulate the expression of components of the antigen processing and presentation pathway, as a common strategy in the evasion of immune surveillance during the induction of cell transformation. Expression of the HPV 18 E7 oncoprotein, like Ad 12 E1A, resulted in repression of the major histocompatibility complex (MHC) class I heavy chain promoter, as well as repression of a bidirectional promoter that regulates expression of the genes encoding the transporter associated with antigen processing subunit 1 (TAP1) and a proteasome subunit, low molecular weight protein 2 (LMP2). HPV 16 E7 also caused a reduction in class I heavy chain promoter activity, however it did not have any significant effect on the activity of the bidirectional promoter. Interestingly, expression of the low-risk HPV 6b E7 protein resulted in an increase in MHC class I heavy chain promoter activity, while repressing the TAP1/LMP2 promoter. Interference with the class I pathway could also explain the ability of low-risk HPVs in inducing benign lesions. PMID- 11039911 TI - Upregulation of Bcl-x and Bfl-1 as a potential mechanism of chemoresistance, which can be overcome by NF-kappaB inhibition. AB - In A549 human lung adenocarcinoma cells, we found that TNF-alpha and several commonly used chemotherapeutic agents upregulated the expression of Bcl-x and/or Bfl-1/A1 through an NF-kappaB-dependent pathway. While parental A549 cells were resistant to the cytotoxic effects of both TNF-alpha and chemotherapy agents, NF kappaB-blocked A549 cells were sensitized to both. Expression of either Bcl-x or Bfl-1/A1 in the NF-kappaB-deficient cells at physiological levels provided differential protection against TNF-alpha and chemotherapeutic treatment. These studies provide a potential mechanism for the phenomenon of chemotherapy-induced chemoresistance, and also reveal a potential strategy by which chemoresistance can be overcome. PMID- 11039913 TI - On fields and fences in science. PMID- 11039912 TI - Identification of negative regulatory regions within the first exon and intron of the BCL6 gene. AB - Chromosomal translocations involving BCL6 gene are frequent in human B-cell lymphomas. Chromosomal breaks preferentially occur within a 3-kb region containing the first exon and intron. Recent reports have revealed that internal deletions or point mutations also are common in this region, suggesting that structural alteration of this region may be a crucial event in the development of lymphomas. In this study, we identified two regions in the BCL6 gene that negatively regulate BCL6 expression. One region, ES, is located within the first exon between nucleotides +472 and +543, and a second region, IS, is located between +783 and + 918 of the first intron. A consensus nucleotide sequence for the binding of the BCL6 protein itself was found within the ES region. An electrophoretic mobility shift assay and a co-transfection experiment using a BCL6 expression vector showed that transcription of the BCL6 gene was negatively regulated by the BCL6 gene product. The IS region which is included in the regions commonly deleted in B-cell lymphomas had a silencer activity. Structural alterations of these two regions may play roles in the deregulated expression of the BCL6 gene in B-cell lymphomas. PMID- 11039914 TI - Comparing clockworks: mouse versus fly. PMID- 11039915 TI - "Rapid" rhythmic discharges of sympathetic nerves: sources, mechanisms of generation, and physiological relevance. AB - Like virtually all other physiological control systems, the sympathetic nervous system controlling cardiovascular function is characterized by the presence of rhythmic activity. These include slow rhythms with frequencies at or below that of the respiration and rapid rhythms with frequencies at or above that of the heart beat. The rapid rhythms are the subject of this review. The specific questions entertained are as follows: (1) Are the rapid cardiac-related and 10-Hz rhythms inherent to central sympathetic networks, or are they imposed on sympathetic nerve discharge (SND) by extrinsic periodic inputs? (2) Does basal SND arise from an anatomically circumscribed "vasomotor center" composed of pacemaker neurons in the rostral ventrolateral medulla or from an anatomically distributed network oscillator composed of different types of brainstem neurons, none of which necessarily have intrinsic pacemaker properties? (3) Are the rapid rhythms generated by single circuits or by systems of coupled oscillators, each with a separate target? (4) Are the rapid rhythms in SND simply by-products of the sympathetic generating mechanisms, or do they subserve selective and special functions, such as the formulation of differential patterns of spinal sympathetic outflow that support particular behaviors? The controversial aspects of these issues and the state-of-the-art analytical methods used to study them are stressed in this review. PMID- 11039916 TI - The effect of different light regimes on adult life span in Drosophila melanogaster is partly mediated through reproductive output. AB - The effects of different light regimes on the fitness of organisms have typically been studied using mean or median adult life span as the sole index of physiological well-being. It is, however, known that life span is inversely related to reproductive output in many species. Moreover, the effects of a given environmental treatment on life span can be due to effects on either age independent mortality or the "rate of aging," or a combination of both. Drawing evolutionary inferences from the effects of light regime on mean or median adult life span alone is difficult and, at best, speculative. We examined the effects of constant light (LL), alternating light-dark cycles (LD 12:12 h), and constant darkness (DD) on the life span of reproducing and virgin flies in four populations of Drosophila melanogaster and also estimated lifetime fecundity in the three light regimes. The light regime effects on life span were further dissected by examining the age-independent mortality and the Gompertz rate of aging under the three light regimes. While mean adult life span of reproducing males and females and virgin females was significantly shorter in LL compared to LD 12:12 h and DD, life-time egg production was highest in LL. Life span of virgin males was not significantly affected by light regime. The rate of aging in reproducing females was higher in LL as compared to DD, whereas age-independent mortality was higher in DD. As reproductive output, especially early in life, is a far more significant contributor to fitness than is life span, our results suggest that the earlier reported deleterious effects of LL on fitness are partly an artifact of examining life span alone, without considering other components of adult fitness that trade off with life span. Our results suggest that detailed investigation of the effects of light regime on the physiological and behavioral processes that accompany reproduction is necessary to fully understand the effects of different light regimes on adult fitness in Drosophila. PMID- 11039917 TI - Light pulses suppress responsiveness within the mouse photic entrainment pathway. AB - We have characterized a decrease in photic responsiveness of the mammalian circadian entrainment pathway caused by light stimulation. Phase delays of the running-wheel activity rhythm were used to quantify the photic responsiveness of the circadian system in mice (C57BL/6J). In an initial experiment, the authors measured the responsiveness to single "saturating" light pulses ("white" fluorescent light; approximately 1876 [microW; 15 min). In two additional experiments, the authors measured responses to this stimulus at several time points following a saturating pulse at CT 14 or CT 16. Data from these experiments were analyzed in two manners. Experiment 2 was analyzed assuming that the phase of the circadian pacemaker was unchanged by an initial pulse, and Experiment 3 was analyzed assuming that the initial pulse induced an instantaneous phase delay. Results reveal a significant reduction in responsivity to light that persists for at least 2 h and possibly up to 4 h after the initial stimulus. Immediately after the stimulus, the responsiveness of the photic entrainment pathway was reduced to levels < or = 12% of normal. After 2 h, the responsiveness was < or = 42% of normal, and by 4 h, responsiveness had recovered to levels that were < or = 60% of normal (levels not statistically different from controls). By the following circadian cycle, responsiveness was more completely recovered, although the magnitude of some phase delays remained < or = 85% of normal. These major reductions in the magnitude of phase delays (and phase response curve amplitude) caused by saturating light pulses confound interpretations of two-pulse experiments designed to measure the rate of circadian phase delays. In addition, the time course for this reduced responsiveness may reflect the time course of cellular and molecular events that underlie light-induced resetting of the mammalian circadian pacemaker. PMID- 11039918 TI - Timing of testicular recrudescence in siberian hamsters is unaffected by pinealectomy or long-day photoperiod after 9 weeks in short days. AB - In this study, the authors asked whether pinealectomy or temporary exposure to a stimulatory photoperiod affects the timing of spontaneous testicular recrudescence in adult Siberian hamsters chronically exposed to short days (9:15 light:dark). In Experiment 1, hamsters were pinealectomized after 6, 9, or 12 weeks in short days. Pinealectomy after 9 or 12 weeks did not affect the timing of spontaneous gonadal growth (27.7 +/- 1.9 and 25.4 +/- 1.3 weeks, respectively) compared to sham-operated controls (28.6 +/- 0.9 weeks). Enlarged testes occurred earlier in animals that were pinealectomized after 6 weeks in short days (21.8 +/ 2.1 weeks). In Experiment 2, adult hamsters were exposed to short days for 9 weeks, transferred to long days (16:8 light:dark) for 4 weeks, and then returned to short days for 23 additional weeks. Although long-day interruption caused gonadal growth in 15 out of 19 hamsters, the temporary long-day exposure did not affect the timing of spontaneous gonadal growth following return to short days (28.2 +/- 0.9 weeks) in 10 of the 15, relative to the timing observed in control hamsters continuously maintained in short days (28.2 +/- 1.1 weeks). Four out of 19 hamsters did not show gonadal growth following long-day exposure. Spontaneous gonadal growth in these hamsters (28.0 +/- 1.4 weeks) also occurred at the same time as controls. The remaining 5 hamsters exhibited enlarged testes following long-day exposure (12.0 +/- 0.0 weeks) but were refractory to the second short day exposure. All hamsters exhibited entrainment of wheel-running activity following the change in photoperiod. A final group of 13 animals were pinealectomized before long-day transfer. They exhibited gonadal growth (at 17.2 +/- 0.8 weeks) but failed to regress a second time when returned to short days. The timing of gonadal growth in these animals was delayed relative to the sham operated hamsters temporarily transferred to long days (Experiment 2) but accelerated relative to the hamsters pinealectomized at 9 weeks, which remained continuously in short days (Experiment 1). The results of both experiments suggest that a pineal-independent process mediates the timing of spontaneous gonadal growth in Siberian hamsters chronically exposed to a short-day photoperiod. PMID- 11039919 TI - Genetic and environmental influences on short-day responsiveness in Siberian hamsters (Phodopus sungorus). AB - Siberian hamsters are photoperiodic rodents that typically exhibit several physiological changes when exposed to a short-day photoperiod. However, development of the winter phenotype in short days is largely conditional on prior photoperiod history: Hamsters that have been reared in an exceptionally long day length (18 L) do not usually exhibit the winter phenotype after transfer to short days, whereas animals reared under "moderately" long days (16 L) are more variable in responsiveness to subsequent short-day exposure, with 20% to 30% generally failing to exhibit winter-type responses. Hamsters reared exclusively in an "intermediate" day length (14 L) are almost uniformly responsive to short photoperiod. In the present study, the authors examine the influence of photoperiod history on short-day responsiveness in a breeding line of hamsters that has been subjected to artificial selection for resistance to the effects of short days. The results demonstrate that photoperiod history is an important determinant of short-day responsiveness in both random-bred (UNS) hamsters and animals artificially selected and bred for nonresponsiveness to short photoperiod (PNR). The PNR hamsters have a reduced requirement for long-day exposure to evoke a state of unresponsiveness to short days. The results are discussed in relation to possible significance for the origin of population and species differences in photoperiod responsiveness. PMID- 11039920 TI - Long photoperiod restores the 24-h rhythm of sleep and EEG slow-wave activity in the Djungarian hamster (Phodopus sungorus). AB - Photoperiod influences the distribution of sleep and waking and electroencephalogram (EEG) power density in the Djungarian hamster. In an experimental procedure combining short photoperiod (SP) and low ambient temperature, the light-dark difference in the amount of sleep was decreased, and the changes in slow-wave activity (SWA) (mean EEG power density between 0.75 and 4.0 Hz) in nonrapid eye movement (NREM) sleep within 24 h were abolished. These findings, obtained in three different groups of animals, suggested that at the lower ambient temperature, the influence of the circadian clock on sleep-wake behavior was diminished. However, it remained unclear whether the changes were due to the photoperiod, ambient temperature, or both. Here, the authors show that EEG and electromyogram recordings in a single group of animals sequentially adapted to a short and long photoperiod (LP) at low ambient temperature (approximately 15 degrees C) confirm that EEG power is reduced in SP. Moreover, the nocturnal sleep-wake behavior and the changes in SWA in NREM sleep over 24 h were restored by returning the animals to LP and retaining ambient temperature at 15 degrees C. Therefore, the effects cannot be attributed to ambient temperature alone but are due to a combined effect of temperature and photoperiod. When the Djungarian hamster adapts to winter conditions, it appears to uncouple sleep regulation from the circadian clock. PMID- 11039921 TI - The human circadian pacemaker can see by the dawn's early light. AB - The authors' previous experiments have shown that dawn simulation at low light intensities can phase advance the circadian rhythm of melatonin in humans. The aim of this study was to compare the effect of repeated dawn signals on the phase position of circadian rhythms in healthy participants kept under controlled light conditions. Nine men participated in two 9-day laboratory sessions under an LD cycle 17.5:6.5 h, < 30:0 lux, receiving 6 consecutive daily dawn (average illuminance 155 lux) or control light (0.1 lux) signals from 0600 to 0730 h (crossover, random-order design). Two modified constant routine protocols before and after the light stimuli measured salivary melatonin (dim light melatonin onset DLMOn and offset DLMOff) and rectal temperature rhythms (midrange crossing time [MRCT]). Compared with initial values, participants significantly phase delayed after 6 days under control light conditions (at least -42 min DLMOn, -54 min DLMOff, -41 min MRCT) in spite of constant bedtimes. This delay was not observed with dawn signals (+10 min DLMOn, +2 min DLMOff, 0 min MRCT). Given that the endogenous circadian period of the human circadian pacemaker is slightly longer than 24 h, the findings suggest that a naturalistic dawn signal is sufficient to forestall this natural delay drift. Zeitgeber transduction and circadian system response are hypothesized to be tuned to the time-rate-of-change of naturalistic twilight signals. PMID- 11039922 TI - Structure of a glycerol-conducting channel and the basis for its selectivity. AB - Membrane channel proteins of the aquaporin family are highly selective for permeation of specific small molecules, with absolute exclusion of ions and charged solutes and without dissipation of the electrochemical potential across the cell membrane. We report the crystal structure of the Escherichia coli glycerol facilitator (GlpF) with its primary permeant substrate glycerol at 2.2 angstrom resolution. Glycerol molecules line up in an amphipathic channel in single file. In the narrow selectivity filter of the channel the glycerol alkyl backbone is wedged against a hydrophobic corner, and successive hydroxyl groups form hydrogen bonds with a pair of acceptor, and donor atoms. Two conserved aspartic acid-proline-alanine motifs form a key interface between two gene duplicated segments that each encode three-and-one-half membrane-spanning helices around the channel. This structure elucidates the mechanism of selective permeability for linear carbohydrates and suggests how ions and water are excluded. PMID- 11039924 TI - Imaging precessional motion of the magnetization vector. AB - We report on imaging of three-dimensional precessional orbits of the magnetization vector in a magnetic field by means of a time-resolved vectorial Kerr experiment that measures all three components of the magnetization vector with picosecond resolution. Images of the precessional mode taken with submicrometer spatial resolution reveal that the dynamical excitation in this time regime roughly mirrors the symmetry of the underlying equilibrium spin configuration and that its propagation has a non-wavelike character. These results should form the basis for realistic models of the magnetization dynamics in a largely unexplored but technologically increasingly relevant time scale. PMID- 11039925 TI - Modulation instability and pattern formation in spatially incoherent light beams. AB - We report on the experimental observation of modulation instability of partially spatially incoherent light beams in noninstantaneous nonlinear media and show that in such systems patterns can form spontaneously from noise. Incoherent modulation instability occurs above a specific threshold that depends on the coherence properties (correlation distance) of the wave packet and leads to a periodic train of one-dimensional filaments. At a higher value of nonlinearity, the incoherent one-dimensional filaments display a two-dimensional instability and break up into self-ordered arrays of light spots. This discovery of incoherent pattern formation reflects on many other nonlinear systems beyond optics. It implies that patterns can form spontaneously (from noise) in diverse nonlinear many-body systems involving weakly correlated particles, such as atomic gases at (or near) Bose-Einstein condensation temperatures and electrons in semiconductors at the vicinity of the quantum Hall regime. PMID- 11039923 TI - Control of viremia and prevention of clinical AIDS in rhesus monkeys by cytokine augmented DNA vaccination. AB - With accumulating evidence indicating the importance of cytotoxic T lymphocytes (CTLs) in containing human immunodeficiency virus-1 (HIV-1) replication in infected individuals, strategies are being pursued to elicit virus-specific CTLs with prototype HIV-1 vaccines. Here, we report the protective efficacy of vaccine elicited immune responses against a pathogenic SHIV-89.6P challenge in rhesus monkeys. Immune responses were elicited by DNA vaccines expressing SIVmac239 Gag and HIV-1 89.6P Env, augmented by the administration of the purified fusion protein IL-2/Ig, consisting of interleukin-2 (IL-2) and the Fc portion of immunoglobulin G (IgG), or a plasmid encoding IL-2/Ig. After SHIV-89.6P infection, sham-vaccinated monkeys developed weak CTL responses, rapid loss of CD4+ T cells, no virus-specific CD4+ T cell responses, high setpoint viral loads, significant clinical disease progression, and death in half of the animals by day 140 after challenge. In contrast, all monkeys that received the DNA vaccines augmented with IL-2/Ig were infected, but demonstrated potent secondary CTL responses, stable CD4+ T cell counts, preserved virus-specific CD4+ T cell responses, low to undetectable setpoint viral loads, and no evidence of clinical disease or mortality by day 140 after challenge. PMID- 11039926 TI - Experimental verification of decoherence-free subspaces. AB - Using spontaneous parametric down-conversion, we produce polarization-entangled states of two photons and characterize them using two-photon tomography to measure the density matrix. A controllable decoherence is imposed on the states by passing the photons through thick, adjustable birefringent elements. When the system is subject to collective decoherence, one particular entangled state is seen to be decoherence-free, as predicted by theory. Such decoherence-free systems may have an important role for the future of quantum computation and information processing. PMID- 11039927 TI - Electronic structure of solids with competing periodic potentials. AB - When electrons are subject to a potential with two incommensurate periods, translational invariance is lost, and no periodic band structure is expected. However, model calculations based on nearly free one-dimensional electrons and experimental results from high-resolution photoemission spectroscopy on a quasi one-dimensional material do show dispersing band states with signatures of both periodicities. Apparent band structures are generated by the nonuniform distribution of electronic spectral weight over the complex eigenvalue spectrum. PMID- 11039928 TI - A stable bicyclic compound with two Si=Si double bonds. AB - In contrast to carbon, silicon does not readily form double bonds, and compounds containing silicon-silicon double bonds can usually be stabilized only by protection with bulky substituents. We have isolated a silicon analog of spiropentadiene 1, a carbon double-ring compound that has not been isolated to date. In the crystal structure of tetrakis[tri(t butyldimethylsilyl)silyl]spiropentasiladiene 2, a substantial deviation from the perpendicular arrangement of the two rings is observed, and the silicon-silicon double bonds are shown to be distorted. Spectroscopic data indicate pronounced interaction between two remote silicon-silicon double bonds in the molecule. Silicon-silicon bonds may be more accessible to synthesis than previously assumed. PMID- 11039929 TI - Dimer preparation that mimics the transition state for the adsorption of H2 on the Si(100)-2 x 1 surface. AB - A chemically induced dimer configuration was prepared on the silicon (Si) (100) surface and was characterized by scanning tunneling microscopy (STM) and spectroscopy (STS). These prepared dimers, which are essentially untilted and differ both electronically and structurally from the dynamically tilting dimers normally found on this surface, are more reactive than normal dimers. For molecular hydrogen (H2) adsorption, the enhancement is about 10(9) at room temperature. There is no appreciable barrier for the H2 reaction at prepared sites, indicating the prepared configuration closely approximates the actual dimer structure in the transition state. This previously unknown ability to prepare specific surface configurations has important implications for understanding and controlling reaction dynamics on semiconductor surfaces. PMID- 11039930 TI - Detection of daily clouds on Titan. AB - We have discovered frequent variations in the near-infrared spectrum of Titan, Saturn's largest moon, which are indicative of the daily presence of sparse clouds covering less than 1% of the area of the satellite. The thermodynamics of Titan's atmosphere and the clouds' altitudes suggest that convection governs their evolutions. Their short lives point to the presence of rain. We propose that Titan's atmosphere resembles Earth's, with clouds, rain, and an active weather cycle, driven by latent heat release from the primary condensible species. PMID- 11039931 TI - Acute sensitivity of landslide rates to initial soil porosity. AB - Some landslides move imperceptibly downslope, whereas others accelerate catastrophically. Experimental landslides triggered by rising pore water pressure moved at sharply contrasting rates due to small differences in initial porosity. Wet sandy soil with porosity of about 0.5 contracted during slope failure, partially liquefied, and accelerated within 1 second to speeds over 1 meter per second. The same soil with porosity of about 0.4 dilated during failure and slipped episodically at rates averaging 0.002 meter per second. Repeated slip episodes were induced by gradually rising pore water pressure and were arrested by pore dilation and attendant pore pressure decline. PMID- 11039932 TI - Rapid evolution of reproductive isolation in the wild: evidence from introduced salmon. AB - Colonization of new environments should promote rapid speciation as a by-product of adaptation to divergent selective regimes. Although this process of ecological speciation is known to have occurred over millennia or centuries, nothing is known about how quickly reproductive isolation actually evolves when new environments are first colonized. Using DNA microsatellites, population-specific natural tags, and phenotypic variation, we tested for reproductive isolation between two adjacent salmon populations of a common ancestry that colonized divergent reproductive environments (a river and a lake beach). We found evidence for the evolution of reproductive isolation after fewer than 13 generations. PMID- 11039933 TI - Natural selection and the reinforcement of mate recognition. AB - Natural selection on mate recognition may often contribute to speciation, resulting in reproductive character displacement. Field populations of Drosophila serrata display reproductive character displacement in cuticular hydrocarbons when sympatric with Drosophila birchii. We exposed field sympatric and allopatric populations of D. serrata to experimental sympatry with D. birchii for nine generations. Cuticular hydrocarbons of field allopatric D. serrata populations evolved to resemble the field sympatric populations, whereas field sympatric D. serrata populations remained unchanged. Our experiment indicates that natural selection on mate recognition resulted in the field pattern of reproductive character displacement. PMID- 11039934 TI - Invasive plants versus their new and old neighbors: a mechanism for exotic invasion. AB - Invading exotic plants are thought to succeed primarily because they have escaped their natural enemies, not because of novel interactions with their new neighbors. However, we find that Centaurea diffusa, a noxious weed in North America, has much stronger negative effects on grass species from North America than on closely related grass species from communities to which Centaurea is native. Centaurea's advantage against North American species appears to be due to differences in the effects of its root exudates and how these root exudates affect competition for resources. Our results may help to explain why some exotic species so successfully invade natural plant communities. PMID- 11039935 TI - Two-amino acid molecular switch in an epithelial morphogen that regulates binding to two distinct receptors. AB - Ectodysplasin, a member of the tumor necrosis factor family, is encoded by the anhidrotic ectodermal dysplasia (EDA) gene. Mutations in EDA give rise to a clinical syndrome characterized by loss of hair, sweat glands, and teeth. EDA-A1 and EDA-A2 are two isoforms of ectodysplasin that differ only by an insertion of two amino acids. This insertion functions to determine receptor binding specificity, such that EDA-A1 binds only the receptor EDAR, whereas EDA-A2 binds only the related, but distinct, X-linked ectodysplasin-A2 receptor (XEDAR). In situ binding and organ culture studies indicate that EDA-A1 and EDA-A2 are differentially expressed and play a role in epidermal morphogenesis. PMID- 11039936 TI - CD95/CD95 ligand interactions on epithelial cells in host defense to Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa causes severe infections, particularly of the lung, that are life threatening. Here, we show that P. aeruginosa infection induces apoptosis of lung epithelial cells by activation of the endogenous CD95/CD95 ligand system. Deficiency of CD95 or CD95 ligand on epithelial cells prevented apoptosis of lung epithelial cells in vivo as well as in vitro. The importance of CD95/CD95 ligand-mediated lung epithelial cell apoptosis was demonstrated by the rapid development of sepsis in CD95- or CD95 ligand-deficient mice, but not in normal mice, after P. aeruginosa infection. PMID- 11039937 TI - Specific mutations induced by triplex-forming oligonucleotides in mice. AB - Triplex-forming oligonucleotides (TFOs) recognize and bind to specific duplex DNA sequences and have been used extensively to modify gene function in cells. Although germ line mutations can be incorporated by means of embryonic stem cell technology, little progress has been made toward introducing mutations in somatic cells of living organisms. Here we demonstrate that TFOs can induce mutations at specific genomic sites in somatic cells of adult mice. Mutation detection was facilitated by the use of transgenic mice bearing chromosomal copies of the supF and cII reporter genes. Mice treated with a supF-targeted TFO displayed about fivefold greater mutation frequencies in the supF gene compared with mice treated with a scrambled sequence control oligomer. No mutagenesis was detected in the control gene (cII) with either oligonucleotide. These results demonstrate that site-specific, TFO-directed genome modification can be accomplished in intact animals. PMID- 11039938 TI - Learning-induced LTP in neocortex. AB - The hypothesis that learning occurs through long-term potentiation (LTP)- and long-term depression (LTD)-like mechanisms is widely held but unproven. This hypothesis makes three assumptions: Synapses are modifiable, they modify with learning, and they strengthen through an LTP-like mechanism. We previously established the ability for synaptic modification and a synaptic strengthening with motor skill learning in horizontal connections of the rat motor cortex (MI). Here we investigated whether learning strengthened these connections through LTP. We demonstrated that synapses in the trained MI were near the ceiling of their modification range, compared with the untrained MI, but the range of synaptic modification was not affected by learning. In the trained MI, LTP was markedly reduced and LTD was enhanced. These results are consistent with the use of LTP to strengthen synapses during learning. PMID- 11039939 TI - Statins: underused by those who would benefit. PMID- 11039940 TI - Another look at visual standards and driving. PMID- 11039941 TI - Headaches after diagnostic dural punctures. PMID- 11039942 TI - Evidence and belief in ADHD. PMID- 11039943 TI - Medical software's free future. PMID- 11039944 TI - UK allows insurers to use gene test for Huntington's disease. PMID- 11039945 TI - Florida jury awards husband damages in wife's smoking death. PMID- 11039946 TI - Tampons could be used to diagnose STDs. PMID- 11039948 TI - In brief PMID- 11039947 TI - Hepatitis C victims sue NHS in class action. PMID- 11039949 TI - EU centre calls for policies to help female drug users. PMID- 11039950 TI - Marijuana has potential for misuse PMID- 11039951 TI - GMC to consult on structural changes. PMID- 11039952 TI - Government asked to withdraw Griffiths report. PMID- 11039953 TI - Fears over anthrax vaccination driving away US reservists. PMID- 11039954 TI - NICE's appraisal procedures attacked. PMID- 11039955 TI - WHO holds hearings on tobacco control. PMID- 11039956 TI - Obesity surgery grows in popularity in the US. PMID- 11039957 TI - India pledges legislation to improve medical services. PMID- 11039959 TI - Women's health study signs up millionth woman. PMID- 11039958 TI - Alliance pledges new cheap TB drug by 2010. PMID- 11039960 TI - Police investigate deaths of terminally ill patients. PMID- 11039961 TI - NHS still rattling tins for funds. PMID- 11039962 TI - Use of lipid lowering drugs for primary prevention of coronary heart disease: meta-analysis of randomised trials. AB - OBJECTIVE: To summarise the effect of primary prevention with lipid lowering drugs on coronary heart disease events, coronary heart disease mortality, and all cause mortality. DESIGN: Meta-analysis. IDENTIFICATION: Systematic search of the Medline database from January 1994 to June 1999 for English language studies examining drug treatment for lipid disorders (use of the MeSH terms "hyperlipidemia" and "anticholesteremic agents," keyword searches for individual drug names, and a search strategy for identifying randomised trials to capture relevant articles); identification of older studies through systematic reviews and hand search of bibliographies. INCLUSION CRITERIA: All randomised trials of at least one year's duration that examined drug treatment for patients with no known coronary heart disease, cerebrovascular disease, or peripheral vascular disease and that measured clinical end points, including all cause mortality, coronary heart disease mortality, and non-fatal myocardial infarctions. DATA EXTRACTION: Review of the articles and extracted relevant data by two authors separately, with disagreements resolved by consensus. RESULTS: Four studies met eligibility criteria. Drug treatment reduced the odds of a coronary heart disease event by 30% (summary odds ratio 0.70, 95% confidence interval 0.62 to 0.79) but not the odds of all cause mortality (0.94, 0.81 to 1.09). When statin drugs were considered alone, no substantial differences in results were found. CONCLUSIONS: Treatment with lipid lowering drugs lasting five to seven years reduces coronary heart disease events but not all cause mortality in people with no known cardiovascular disease. PMID- 11039963 TI - Randomised controlled trial of atraumatic versus standard needles for diagnostic lumbar puncture. AB - OBJECTIVE: To compare the ease of use of atraumatic needles with standard needles for diagnostic lumbar puncture and the incidence of headache after their use. DESIGN: Double blind, randomised controlled trial. SETTING: Investigation ward of a neurology unit in a university hospital. PARTICIPANTS: 116 patients requiring elective diagnostic lumbar puncture. INTERVENTIONS: Standardised protocol for lumbar puncture with 20 gauge atraumatic or standard needles. OUTCOME MEASURES: The primary end point was intention to treat analysis of incidence of moderate to severe headache, assessed at one week by telephone interview. Secondary end points were incidence of headache at one week analysed by needle type, ease of use by operator according to a visual analogue scale, incidence of backache, and failure rate of puncture. RESULTS: Valid outcome data were available for 97 of 101 patients randomised. Baseline characteristics were matched except for higher body mass index in the standard needle group. By an intention to treat analysis the absolute risk of moderate to severe headache with atraumatic needles was reduced by 26% (95% confidence interval 6% to 45%) compared with standard needles, but there was a non-significantly greater absolute risk of multiple attempts at lumbar puncture (14%, -4% to 32%). Higher body mass index was associated with an increased failure rate with atraumatic needles, but the reduced incidence of headache was maintained. The need for medical interventions was reduced by 20% (1% to 40%). CONCLUSIONS: Atraumatic needles significantly reduced the incidence of moderate to severe headache and the need for medical interventions after diagnostic lumbar punctures, but they were associated with a higher failure rate than standard needles. PMID- 11039964 TI - Reliability of Snellen charts for testing visual acuity for driving: prospective study and postal questionnaire. AB - OBJECTIVES: To assess the ability of patients with binocular 6/9 or 6/12 vision on the Snellen chart (Snellen acuity) to read a number plate at 20.5 m (the required standard for driving) and to determine how health professionals advise such patients about driving. DESIGN: Prospective study of patients and postal questionnaire to healthcare professionals. SUBJECTS: 50 patients with 6/9 vision and 50 with 6/12 vision and 100 general practitioners, 100 optometrists or opticians, and 100 ophthalmologists. SETTING: Ophthalmology outpatient clinics in Sheffield. MAIN OUTCOME MEASURES: Ability to read a number plate at 20.5 m and health professionals' advice about driving on the basis of visual acuity. RESULTS: 26% of patients with 6/9 vision failed the number plate test, and 34% with 6/12 vision passed it. Of the general practitioners advising patients with 6/9 vision, 76% said the patients could drive, 13% said they should not drive, and 11% were unsure. Of the general practitioners advising patients with 6/12 vision, 21% said the patients could drive, 54% said they should not drive, and 25% were unsure. The level of acuity at which optometrists, opticians, and ophthalmologists would advise drivers against driving ranged from 6/9(-2) (ability to read all except two letters on the 6/9 line of the Snellen chart) to less than 6/18. CONCLUSIONS: Snellen acuity is a poor predictor of an individual's ability to meet the required visual standard for driving. Patients with 6/9 vision or less should be warned that they may fail to meet this standard, but those with 6/12 vision should not be assumed to be below the standard. PMID- 11039966 TI - Never has a three hour flight seemed so long PMID- 11039965 TI - Unjustified exclusion of elderly people from studies submitted to research ethics committee for approval: descriptive study. PMID- 11039967 TI - Effects of a programme of multifactorial home visits on falls and mobility impairments in elderly people at risk: randomised controlled trial. AB - OBJECTIVE: To evaluate whether a programme of multifactorial home visits reduces falls and impairments in mobility in elderly people living in the community. DESIGN: Randomised controlled trial with 18 months of follow up. SETTING: Six general practices in Hoensbroek, the Netherlands. PARTICIPANTS: 316 people aged 70 and over living in the community, with moderate impairments in mobility or a history of recent falls. INTERVENTION: Five home visits by a community nurse over a period of one year. Visits consisted of screening for medical, environmental, and behavioural factors causing falls and impairments in mobility, followed by specific advice, referrals, and other actions aimed at dealing with the observed hazards. MAIN OUTCOME MEASURES: Falls and impairments in mobility. RESULTS: No differences were found in falls and mobility outcomes between the intervention and usual care groups. CONCLUSION: Multifactorial home visits had no effects on falls and impairments in mobility in elderly people at risk who were living in the community. Because falls and impairments in mobility remain a serious problem among elderly people, alternative strategies should be developed and evaluated. PMID- 11039968 TI - Rectal bleeding and colorectal cancer in general practice: diagnostic study. PMID- 11039969 TI - Untied PMID- 11039970 TI - Lesson of the week: tracheal stenosis after intubation. PMID- 11039972 TI - Only one publication! PMID- 11039971 TI - Partial seizures presenting as panic attacks. PMID- 11039973 TI - ABC of colorectal cancer: screening. PMID- 11039974 TI - Guidelines for the prevention of falls in people over 65. The Guidelines' Development Group. PMID- 11039975 TI - Managed care of chronically ill older people: the US experience. PMID- 11039976 TI - The NHS plan. Plan represents considerable privatisation. PMID- 11039977 TI - The NHS plan. Plan puts patients first, but who will want to care for them? PMID- 11039978 TI - The NHS plan. BMA representatives must negotiate for the NHS as it will be. PMID- 11039979 TI - Private finance scheme for Worcester. Journal has not presented balanced picture of modern health planning. PMID- 11039980 TI - Meta-analysis of increased inhaled steroid or addition of salmeterol in asthma. Greening et al's study should not have been included. PMID- 11039981 TI - Meta-analysis of increased inhaled steroid or addition of salmeterol in asthma. Study should have been more thorough. PMID- 11039982 TI - Arterial and venous disease and deep vein thrombosis. Timing of thromboprophylaxis for general surgery should be discussed with anaesthetists. PMID- 11039983 TI - Arterial and venous disease and deep vein thrombosis. Injecting drug use is major risk factor for deep vein thrombosis. PMID- 11039984 TI - Babies sleeping with parents and sudden infant death syndrome. Invoking sudden infant death syndrome in cosleeping may be misleading. PMID- 11039985 TI - Global medical knowledge database. IT, or not IT? PMID- 11039986 TI - Global medical knowledge database. New professional obligation arises. PMID- 11039987 TI - Isolated systolic hypertension. Life assurance industry highlights importance of blood pressure control. PMID- 11039988 TI - Disability one year after head injury. Role of alcohol needs to be examined. PMID- 11039989 TI - When people would choose treatment has wide implications. PMID- 11039990 TI - Anaesthesia is also risky in patients with porphyria. PMID- 11039991 TI - Presumed consent further undermines medical ethics. PMID- 11039992 TI - Eating disorders are becoming more common in the East too. PMID- 11039993 TI - Refugee doctors PMID- 11039994 TI - Glagovian remodelling, plaque composition, and stenosis generation. PMID- 11039995 TI - Stamps in cardiology: Institutions. PMID- 11039996 TI - Genetics of dilated cardiomyopathy: a molecular maze? PMID- 11039997 TI - Images in cardiology: Cardiac hydatid cyst. PMID- 11039998 TI - Cellular cardiomyoplasty: a new hope in heart failure? PMID- 11039999 TI - Radionuclide investigation of congenital heart disease. PMID- 11040000 TI - Stent magic! The genie has escaped from the bottle. PMID- 11040002 TI - Hypertrophic cardiomyopathy: the interrelation of disarray, fibrosis, and small vessel disease. AB - OBJECTIVE: To make a quantitative assessment of the relation between disarray, fibrosis, and small vessel disease in hypertrophic cardiomyopathy. DESIGN: Detailed macroscopic and histological examination at 19 segments of the left and right ventricle and the left atrial free wall. PATIENTS: 72 patients with hypertrophic cardiomyopathy who had suffered sudden death or progression to end stage cardiac failure (resulting in death or heart transplantation). MAIN OUTCOME MEASURES: The presence of scarring, atrial dilatation, and a mitral valve impact lesion were noted, and heart weight, wall thickness, per cent disarray, per cent fibrosis, and per cent small vessel disease quantitated for each heart. RESULTS: Within an individual heart the magnitude of hypertrophy correlated with the severity of fibrosis (p = 0.006) and disarray (p = 0.0002). Overall, however, total heart weight related weakly but significantly to fibrosis (r = 0.4, p = 0.0001) and small vessel disease (r = 0.3, p = 0.03), but not to disarray. Disarray was greater in hearts with mild left ventricular hypertrophy (maximum wall thickness < 20 mm) and preserved systolic function (60.9 (26)% v 43 (20.4)% respectively, p = 0.02) and hearts without a mitral valve impact lesion (26.3% v 18.9%, p = 0.04), but was uninfluenced by sex. Fibrosis was influenced by sex (7% in male patients and 4% in female, p = 0.04), but not by the presence of an impact lesion. No relation was found between disarray, fibrosis, and small vessel disease. CONCLUSIONS: Myocyte disarray is probably a direct response to functional or structural abnormalities of the mutated sarcomeric protein, while fibrosis and small vessel disease are secondary phenomena unrelated to disarray, but modified by factors such as left ventricular mass, sex, and perhaps local autocrine factors. PMID- 11040001 TI - New recipes for in-stent restenosis: cut, grate, roast, or sandwich the neointima? PMID- 11040003 TI - Visualisation of systolic myocardial perfusion defect induced by septal squeezing. PMID- 11040004 TI - Size of emptied plaque cavity following spontaneous rupture is related to coronary dimensions, not to the degree of lumen narrowing. A study with intravascular ultrasound in vivo. AB - OBJECTIVE: To identify any potential relations between the size of an emptied plaque cavity and the remodelling pattern, plaque or vessel dimensions, lumen narrowing, and other ultrasonic lesion characteristics. DESIGN: Intravascular ultrasound was used to examine prospectively 51 ruptured ulcerated coronary plaques. Cross sectional area measurements comprised lumen, vessel, plaque, and emptied plaque cavity. Lumen narrowing was calculated as 1 - (lesion lumen area/reference lumen area) x 100%. A remodelling index was calculated as lesion vessel area/reference vessel area, and plaques were divided into those with values > 1.05 (group A) and /= 15% (p = 0.01). CONCLUSIONS: Progression of CAVD is characterised by a continuing increase in intimal hyperplasia, especially within the first year after heart transplantation. LDL cholesterol is an important predictor of major progression. PMID- 11040008 TI - Natural and unnatural history of pulmonary atresia. AB - OBJECTIVE: To investigate mortality, cause of death, survival, and quality of life in all types of cardiac malformation with congenital pulmonary atresia. DESIGN: Retrospective analysis. SETTING: The resident population of one health region with a single tertiary referral centre. PATIENTS: All babies with pulmonary atresia live born in 1980 to 1995. MAIN OUTCOME MEASURES: Anatomical classification, total mortality, cause of death, duration of survival, exercise ability. All cases were classified as pulmonary atresia with intact septum (PA IVS), pulmonary atresia with ventricular septal defect (PA-VSD), or pulmonary atresia with complex cardiac malformation (complex pulmonary atresia). RESULTS: 129 cardiac malformations with congenital pulmonary atresia were identified from 601 635 live births (21.4/100 000): 29 had PA-IVS, 60 had PA-VSD, and 40 had complex pulmonary atresia. Total mortality was 72/129 (56%), with 15 deaths in the first week and 49 in the first year. There were 23 surgical deaths, 33 hospital deaths (not related to surgery), and 16 sudden deaths, 12 of which remained unexplained. The sudden death rate was 29/1000 patient years of follow up. Of the 57 survivors, 39% have exercise ability I or II and 61% III or IV. Definitive surgical repair produced better exercise ability. CONCLUSIONS: Early mortality is high in all types of pulmonary atresia, although survival has improved in recent years. Most children who have not undergone definitive repair have significant exercise limitation. PMID- 11040009 TI - Multiple milking in hypertrophic cardiomyopathy. PMID- 11040010 TI - Time dependent variability of QT dispersion after acute myocardial infarction and its relation to ventricular fibrillation: a prospective study. AB - OBJECTIVE: To show whether increased QT dispersion on admission predicts ventricular fibrillation after acute myocardial infarction, and to determine the nature of time related changes in QT dispersion. DESIGN: Prospective cohort study. SETTING: Coronary care units of three teaching hospitals in Newcastle-upon Tyne over an eight month period. PATIENTS: All had acute myocardial infarction according to World Health Organization criteria. INTERVENTIONS: For all patients, QT dispersion (QTd) and Bazett rate corrected QTc dispersion (QTcd) were measured from a high quality 12 lead ECG recorded on admission at a paper speed of 50 mm/s. In a subset, serial ECGs were recorded regularly to show time related changes in QTcd following acute myocardial infarction. MAIN OUTCOME MEASURES: Occurrence of ventricular fibrillation within the first 24 hours after myocardial infarction. RESULTS: Data collected from 201 patients, 12 of whom (6%) developed ventricular fibrillation within 24 hours. Neither QTd nor QTcd differed between those developing ventricular fibrillation and those who did not: QTd mean (SD), 74 (24) ms (95% confidence interval (CI) 59 to 89) v 66 (24) ms (95% CI 62 to 70), respectively; QTcd, 86 (26) ms(0.5) (95% CI 70 to 102) v 77 (29) ms(0.5) (95% CI 72 to 82), respectively. Significant QTcd changes occurred early after myocardial infarction. CONCLUSIONS: Admission QTd and QTcd do not predict ventricular fibrillation after acute myocardial infarction. There are significant changes in QTcd with time, which may account for this measured lack of correlation. PMID- 11040011 TI - The normal response to prolonged passive head up tilt testing. AB - OBJECTIVE: To define the responses to head up tilt in a large group of normal adult subjects using the most widely employed protocol for tilt testing. METHODS: 127 normal subjects aged 19-88 years (mean (SD), 49 (20) years) without a previous history of syncope underwent tilt testing at 60 degrees for 45 minutes or until syncope intervened. Blood pressure monitoring was performed with digital photoplethysmography, providing continuous, non-invasive, beat to beat heart rate and pressure measurements. RESULTS: 13% of subjects developed vasovagal syncope after a mean (SD) tilt time of 31.7 (12. 4) minutes (range 8.5-44.9 minutes). Severe cardioinhibition during syncope was observed less often than is reported in patients investigated for syncope. There were no differences in the age or sex distributions of subjects with positive or negative outcomes, or in the proportions with cardioinhibitory and vasodepressor vasovagal syncope compared with previously reported patient populations. Subjects with negative outcomes showed age related differences in heart rate and blood pressure behaviour throughout tilt. CONCLUSIONS: False positive results with tilting appear to be common. This has important implications for the use of diagnostic tilt testing. The magnitude of the heart rate and blood pressure changes observed during negative tilts largely invalidates previously suggested criteria for abnormal non syncopal outcomes. PMID- 11040012 TI - Isolated left atrial tamponade following circumflex artery angioplasty. PMID- 11040013 TI - Does primary stenting preserve cardiac function in myocardial infarction? A case control study. NORTH-981 investigators. Network of revascularisation therapy in Hokkaido. AB - OBJECTIVE: To investigate whether coronary stenting limits myocardial injury and preserves left ventricular function. DESIGN AND SETTING: Prospective multicentre case-control study of primary percutaneous transluminal coronary angioplasty (PTCA) with and without stenting, performed in seven cardiovascular centres. SUBJECTS AND METHODS: 45 consecutive patients with acute myocardial infarction who were treated with successful primary stenting (Stent group) and did not have restenosis were paired with 45 matched control subjects with acute myocardial infarction treated by successful primary PTCA without stenting, also with no restenosis (POBA group). RESULTS: In comparison with the POBA group, the Stent group-especially those patients with a left anterior descending coronary artery lesion-had a smaller hypokinesis area (mean (SD): 15. 1 (20.0) v 34.4 (24.3) chords), reduced hypokinesis area/risk area (25.2 (31.9)% v 58.8 (40.1)%), and a larger ejection fraction (63.3 (10.2)% v 51.7 (11.7)%) evaluated by quantitative left ventriculography using the centerline method. In the Stent group, the correlation between risk area and hypokinesis area was significantly shifted downward. Multiple logistic regression analysis on infarct size limitation (hypokinesis area/risk area < 50%) identified preinfarction angina in all subjects and preinfarction angina and stenting in patients with left anterior descending coronary artery lesions as explanatory factors. CONCLUSIONS: Primary PTCA using a coronary stent is effective in preventing myocardial injury and restoring left ventricular function in patients with anterior acute myocardial infarction. PMID- 11040014 TI - Is provisional stenting the effective option? The WIDEST study (Wiktor stent in de novo stenosis). Widest Trial Investigators' Group. AB - AIM: To compare the immediate and late outcomes of patients treated by a policy of routine stent implantation with routine balloon angioplasty and the use of stents only when an ideal result has not been obtained. METHODS: A nine centre, multinational, randomised study of 300 patients with coronary artery disease thought suitable for treatment of a single lesion by balloon angioplasty or stent implantation. Only new lesions in patients who had not undergone previous bypass surgery were included, and totally occluded vessels were excluded. RESULTS: The initial procedure was considered successful in 96% of patients. There was more complete angiographic restoration of luminal diameter in patients treated by elective stent (minimum lumen diameter (MLD) 2.68 mm for stent v 2.27 mm for balloon; p < 0.007), but analysis of the subgroup of balloon angioplasty patients who crossed over to stenting showed that they achieved similar results to the elective stent group. Late luminal loss was greater in stented patients than in those undergoing balloon angioplasty only, and by six months the angiographic benefit of stenting had disappeared (MLD 1.90 mm for stent group v 2.00 mm for balloon angioplasty). Angiographic and clinical results in the balloon angioplasty group were assisted by the high crossover rate (30.1%). Both groups had similar symptom relief, with 58.9% of patients improving by two or more angina grades. The need for further revascularisation was also similar in the two groups at one year (18.2% in the stented group v 17.1% in the balloon angioplasty group). Haemorrhagic complications at the local arterial entry site were more common than expected and were distributed equally between the patients receiving full anticoagulation and those receiving antiplatelet treatment only. The results of both Wiktor stent placement and balloon angioplasty were similar to the findings in the stent group in previous randomised studies (Benestent II, STRESS). CONCLUSIONS: Provisional stenting appears to offer the same longer term outcome as elective stenting in this selected group of patients. Improvement in the results of conventional balloon angioplasty in the past 10 years means that a policy of obtaining an ideal result without the use of stents appears to be practicable in many of these patients, with consequent cost savings. PMID- 11040016 TI - Images in cardiology: Mitral valve aneurysm. PMID- 11040015 TI - Complex stenosis morphology and vasomotor responses to inhibition of nitric oxide synthesis. AB - OBJECTIVE: To assess the relation between coronary vasomotor effects of N(G) monomethyl-L-arginine (LNMMA) administration and coronary stenosis morphology, length, and severity in patients with stable angina. DESIGN: In 28 patients (24 male, four female) with coronary artery disease and chronic stable angina, intracoronary normal saline and 4 micromol/min LNMMA were infused for four minutes each, followed by an intracoronary bolus of 250 microg glyceryl trinitrate. Coronary stenoses were classified as concentric (smooth), eccentric (smooth), or complicated (irregular). The diameters of these stenoses and their adjacent reference proximal segments were measured by quantitative angiography. RESULTS: During LNMMA infusion a significantly larger proportion of complicated stenoses than concentric and eccentric stenoses constricted by >/= 5% (p < 0.01) and the magnitude of vasoconstriction was greater in complicated than in concentric and eccentric stenoses (p < 0.05). For complicated stenoses the magnitude of constriction (in mm) with reference to normal saline was greater than that of the concentric and eccentric stenoses (p < 0.05), whereas concentric and eccentric stenoses constricted similarly. Irrespective of the type of morphology, there was a correlation (p < 0.05) between both the severity and the length of stenoses and the magnitude of vasoconstriction to LNMMA. A similar proportion of concentric, eccentric, and complicated stenoses showed >/= 5% increase in diameter with glyceryl trinitrate, and the magnitude of the response was similar in the three groups. CONCLUSIONS: In patients with coronary artery disease, the response to LNMMA is greater when stenosis morphology is complex, indicating greater nitric oxide activity. This provides further evidence that plaques with complex morphology are in an active state. PMID- 11040017 TI - Health related quality of life after conservative or invasive treatment of inducible postinfarction ischaemia. DANAMI study group. AB - OBJECTIVE: To assess health related quality of life in patients with inducible postinfarction ischaemia. DESIGN: A questionnaire based follow up study on patients randomised to conservative or invasive treatment because of postinfarction ischaemia. SETTING: Seven county hospitals in eastern Denmark and the Heart Centre, National University Hospital, Copenhagen, Denmark. PATIENTS: 113 patients with inducible postinfarction ischaemia: 51 were randomised to conservative treatment and 62 to invasive treatment. Average follow up time was three years (19-57 months). MAIN OUTCOME MEASURES: SF-36, Rose angina and dyspnoea questionnaire, drug use, lifestyle, and cognitive function. RESULTS: Invasively treated patients scored better on the SF-36 scales of physical functioning (p = 0.03) and on role-physical (p = 0.04) and physical component scales (p = 0.05) and took significantly less anti-ischaemic drug treatment. Angina occurred in 18% of the invasively treated patients and 31% of the conservatively treated patients (p = 0.09). However, more invasively treated patients suffered from concentration difficulties (18% v 4%; p = 0.04). CONCLUSIONS: Patients who were treated invasively had better health related quality of life scores in the physical variables compared with conservatively treated patients. However, a larger proportion of invasively treated patients had concentration difficulties. PMID- 11040019 TI - Polymorphisms of the P-selectin gene and risk of myocardial infarction in men and women in the ECTIM extension study. Etude cas-temoin de l'infarctus myocarde. AB - BACKGROUND AND OBJECTIVE: Studies in animal models and humans implicate cell adhesion molecules in atherogenesis but their role in mediating the risk of myocardial infarction is unclear. The ECTIM (etude cas-temoin de l'infarctus myocarde) extension study was established to determine whether a previously implicated polymorphism of the P-selectin gene was associated with myocardial infarction risk in men and women in Belfast and Glasgow. PATIENTS AND STUDY SETTING: 696 cases with a recent myocardial infarction and 561 age matched controls (both male and female) were recruited into a case-control study in MONICA project areas of Belfast and Glasgow. METHODS: Demographic and lifestyle information was collected by interview administered questionnaire, and each subject was examined and provided a blood sample for DNA extraction. The polymerase chain reaction (PCR) was used to amplify regions encompassing the P selectin Thr-->Pro (A/C) polymorphism at position 715. Genotype odds ratios for myocardial infarction were estimated by logistic regression adjusted for population, age, and sex. RESULTS: There was no significant association between conventional risk factors (such as hypercholesterolaemia, increased body mass index, or raised blood pressure) and either the rare or the common Pro(715) allele of the P-selectin gene in controls. Overall, comparing Pro(715)/Pro(715) and Pro(715)/Thr(715) with Thr(715)/Thr(715), with adjustment for centre, age, and sex, the odds ratio was 0.78 (95% confidence interval 0.60 to 1.00) (p = 0.054), indicating a "protective" effect of the less common Pro(715) allele. There was no significant heterogeneity in odds ratios between men and women either in this sample or when combined with the original ECTIM subjects. CONCLUSIONS: In a large population based study in two regions of the UK, we have been able to corroborate the earlier ECTIM findings of a lower frequency of the Thr/Pro(715) polymorphism in subjects with myocardial infarction. An apparently "protective effect" of similar magnitude also seems to apply to women. PMID- 11040018 TI - Haemochromatosis gene mutations in idiopathic dilated cardiomyopathy. AB - BACKGROUND: Two common mutations of the haemochromatosis associated gene (HFE) (cys282tyr (C282Y) and his63asp (H63D)) have been implicated in haemochromatosis and as modulators in cardiovascular disease. OBJECTIVE: To investigate the role of these mutations in the pathogenesis of idiopathic dilated cardiomyopathy. DESIGN AND SETTING: Case-control and prospective cohort study of patients attending a cardiomyopathy unit in a tertiary referral cardiac centre. METHODS: 207 unrelated white patients with dilated cardiomyopathy, followed up for 259 patient years, and 200 controls were tested for HFE C282Y and H63D mutations by polymerase chain reaction and restriction digestion. RESULTS: 31/207 patients (15%) v 24/200 controls (12%) carried C282Y (adjusted odds ratio (OR) 1.2 (95% confidence interval 0.7 to 2.2)), 74/207 (36%) v 53/200 (27%) carried H63D (OR 1.6 (1.1 to 2.5)), and 10/207 (4.8%) v 4/200 (2%) were compound heterozygotes (OR 2.6 (0.8 to 8.5)). Four patients and six controls were H63D homozygous and one was C282Y homozygous. There was a progressive increase in mean serum iron ([Fe]) and transferrin saturations from patients with no mutation ([Fe] = 16.3 micromol/l, transferrin saturation = 23.7%) through H63D heterozygotes (17.5 micromol/l, 25.8%), C282Y heterozygotes (17.1 micromol/l, 26.6%), H63D homozygotes (20.0 micromol/l, 33.5%), compound heterozygotes (26.8 micromol/l, 41.7%), and C282Y homozygotes (34 micromol/l, 71%). At follow up (median 90 months) the rate of death or cardiac transplantation was 52/207 (25%). C282Y heterozygotes had less ventricular dilatation (mean (SD): 59.9 (1.7) mm v 64.9 (0.9) mm, p < 0.05), better fractional shortening (24 (1. 7)% v 18.8 (1.4)%, p < 0.01), and a trend towards improved survival without transplantation. [Fe] and transferrin saturation did not correlate with disease severity and were not associated with reduced survival. CONCLUSIONS: The frequency of the H63D mutation is significantly increased in patients with idiopathic dilated cardiomyopathy. As H63D has a relatively minor effect on iron status, the mechanism of this association may be unrelated to iron metabolism. PMID- 11040020 TI - How to survive myocardial rupture after myocardial infarction. PMID- 11040021 TI - Radiofrequency catheter ablation of ventricular tachycardia. PMID- 11040022 TI - Heart disease in the elderly. PMID- 11040023 TI - Anticoagulation in valvar heart disease: new aspects and management during non cardiac surgery. PMID- 11040024 TI - Exercise induced supraventricular tachycardia? PMID- 11040025 TI - Novel expression of VCAM-1 on the mitral valve in a patient with primary antiphospholipid antibody syndrome. PMID- 11040028 TI - Three dimensional colour Doppler echocardiography for the characterisation and quantification of cardiac flow events. PMID- 11040029 TI - Three dimensional echocardiography for the assessment of mitral valve disease. PMID- 11040030 TI - Tissue Doppler imaging: current and potential clinical applications. PMID- 11040031 TI - A survey of sedation and monitoring practices during transoesophageal echocardiography in the UK: are recommended guidelines being followed? PMID- 11040033 TI - Differential modes of agonist binding to 5-hydroxytryptamine(2A) serotonin receptors revealed by mutation and molecular modeling of conserved residues in transmembrane region 5. AB - Site-directed mutagenesis and molecular modeling were used to investigate the molecular interactions involved in ligand binding to, and activation of, the rat 5-hydroxytryptamine(2A) (5-HT(2A)) serotonin (5-HT) receptor. Based on previous modeling studies utilizing molecular mechanics energy calculations and molecular dynamics simulations, four sites (S239[5.43], F240[5.44], F243[5.47], and F244[5.48]) in transmembrane region V were selected, each predicted to contribute to agonist and/or antagonist binding. The F243A mutation increased the affinity of (+/-)4-iodo-2, 5-dimethoxyphenylisopropylamine, decreased the binding of alpha methyl-5HT, N-omega-methyl-5HT, ketanserin, ritanserin, and spiperone and had no effect on the binding of 5-HT and 5-methyl-N, N-dimethyltryptamine. The F240A mutant had no effect on the binding of any of the ligands tested, whereas F244A caused an agonist-specific decrease in binding affinity (3- to 10-fold). S239A caused a 6- to 13-fold decrease in tryptamine-binding affinity and a 5-fold increase in affinity of 4-iodo-2, 5-dimethoxyphenylisopropylamine. A subset of the agonists used in binding studies were used to determine the efficacies and potencies of these mutants to activate phosphoinositide hydrolysis. The F243A and F244A mutations reduced agonist stimulated phosphoinositide hydrolysis, whereas the S239A and F240A mutations had no effect. There was little correlation between agonist binding and second messenger production. Furthermore, molecular dynamics simulations, considering these data, produced ligand-bound structures utilizing substantially different bonding interactions even among structurally similar ligands (differing by as little as one methyl group). Taken together, these results suggest that relatively minor changes in either receptor or ligand structure can produce drastic and unpredictable changes in both binding interactions and 5-HT(2A) receptor activation. Thus, our finding may have major implications for the future and feasibility of receptor structure-based drug design. PMID- 11040034 TI - Spontaneous activation of beta(2)- but not beta(1)-adrenoceptors expressed in cardiac myocytes from beta(1)beta(2) double knockout mice. AB - Although ligand-free, constitutive beta(2)-adrenergic receptor (AR) signaling has been demonstrated in naive cell lines and in transgenic mice overexpressing cardiac beta(2)-AR, it is unclear whether the dominant cardiac beta-AR subtype, beta(1)-AR, shares the ability of spontaneous activation. In the present study, we expressed human beta(1)- or beta(2)-AR via recombinant adenoviral infection in ventricular myocytes isolated from beta(1)beta(2)-AR double knockout mice, creating pure beta(1)-AR and beta(2)-AR systems with variable receptor densities. A contractile response to a nonselective beta-AR agonist, isoproterenol, was absent in double knockout mouse myocytes but was fully restored after adenoviral beta(1)-AR or adenoviral beta(2)-AR infection. Increasing the titer of adenoviral vectors (multiplicity of infection 10-1000) led to a dose-dependent expression of beta(1)- or beta(2)-AR with a maximal density of 1207 +/- 173 (36-fold over the wild-type control value) and 821+/-38 fmol/mg protein (69-fold), respectively. Using confocal immunohistochemistry, we directly visualized the cellular distribution of beta(1)-AR and beta(2)-AR and found that both subtypes were distributed on the cell surface membrane and transverse tubules, resulting in a striated pattern. In the absence of ligand, beta(2)-AR expression resulted in graded increases in baseline cAMP and contractility up to 428% and 233% of control, respectively, at the maximal beta(2)-AR density. These effects were specifically reversed by a beta(2)-AR inverse agonist, ICI 118,551 (10(-7) M). In contrast, overexpression of beta(1)-AR, even at a greater density, failed to enhance either basal cAMP or contractility; the alleged beta(1)-AR inverse agonist, CGP 20712A (10(-6) M), had no significant effect on basal contraction in these cells. Thus, we conclude that acute beta(2)-AR overexpression in cardiac myocytes elicits significant physiological responses due to spontaneous receptor activation; however, this property is beta-AR subtype specific because beta(1)-AR does not exhibit agonist-independent spontaneous activation. PMID- 11040035 TI - Functional comparisons of the lysophosphatidic acid receptors, LP(A1)/VZG-1/EDG 2, LP(A2)/EDG-4, and LP(A3)/EDG-7 in neuronal cell lines using a retrovirus expression system. AB - Lysophosphatidic acid (LPA) is a potent lipid mediator with diverse physiological actions on a wide variety of cells and tissues. Three cognate G-protein-coupled receptors have been identified as mammalian LPA receptors: LP(A1)/VZG-1/EDG-2, LP(A2)/EDG-4, and LP(A3)/EDG-7. The mouse forms of these genes were analyzed in rodent cell lines derived from nervous system cells that can express these receptors functionally. An efficient retrovirus expression system was used, and each receptor was heterologously expressed in B103 rat neuroblastoma cells that neither express these receptors nor respond to LPA in all assays tested. Comparative analyses of signaling pathways that are activated within minutes of ligand delivery were carried out. LPA induced cell rounding in LP(A1)- and LP(A2) expressing cells. By contrast, LP(A3) expression resulted in neurite elongation in B103 cells and inhibited LPA-dependent cell rounding in TR mouse neuroblast cells that endogenously express LP(A1) and LP(A2) but not LP(A3). Each of the receptors could couple to multiple G-proteins and induced LPA-dependent inositol phosphate production, mitogen-activated protein kinase activation, and arachidonic acid release while inhibiting forskolin-induced cAMP accumulation, although the efficacy and potency of LPA varied from receptor to receptor. These results indicate both shared and distinct functions among the three mammalian LPA receptors. The retroviruses developed in this study should provide tools for addressing these functions in vivo. PMID- 11040036 TI - Cyclic AMP and protein kinase A stimulate Cdc42: role of A(2) adenosine receptors in human mast cells. AB - The functional activity of Cdc42 is known to be regulated by proteins that control its GDP/GTP-bound state. However, there is still limited information on how Cdc42 is controlled by G-protein-coupled receptors. Adenosine receptors belong to the G-protein-coupled receptor family of cell surface receptors. Human HMC-1 mast cells express the high-affinity A(2A) and the low-affinity A(2B) subtypes of adenosine receptors known to increase intracellular cAMP levels. We found that both subtypes of A(2) adenosine receptors activate Cdc42 in HMC-1 cells. Furthermore, stimulation of adenylate cyclase with forskolin, or loading of HMC-1 with the cell-permeable cAMP analog 8-Br-cAMP, activated Cdc42. Stimulation of Cdc42 by cAMP was also observed in CHO-K1 and COS-7 cells. Protein kinase A (PKA)-mediated phosphorylation is likely involved in cAMP-dependent Cdc42 activation, because transient expression of the PKA catalytic subunit in COS-7 cells activated Cdc42. Inhibition of protein phosphatases 1 and 2A with calyculin A potentiated the effects of 5'-N-ethylcarboxamidoadenosine and 8-Br cAMP, whereas the selective PKA inhibitor H-89 reversed the activation of Cdc42. We demonstrated that Cdc42 is a poor substrate for PKA phosphorylation in vitro and in intact cells. Our data suggest that PKA does not phosphorylate Cdc42 directly. Instead, the proteins that modulate the GDP/GTP-bound state of Cdc42 may be the primary targets of PKA phosphorylation. PMID- 11040037 TI - Structural insights into the amino-terminus of the secretin receptor: I. Status of cysteine and cystine residues. AB - The secretin receptor is prototypic of the class II family of G protein-coupled receptors, with a long extracellular amino-terminal domain containing six highly conserved Cys residues and one Cys residue (Cys(11)) that is present only in the most closely related family members. This domain is critical for function, with some component Cys residues believed to be involved in key disulfide bonds, although these have never been directly demonstrated. Here, we examine the functional importance of each of these residues and determine their involvement in disulfide bonds. Secretin binding was markedly diminished after treating cells with cell-impermeant reducing reagents, supporting the presence of important extracellular disulfide bonds. To determine whether the amino-terminal domain was covalently attached to the receptor body by disulfide linkage, a strategy was implemented that involved introduction of an acid-labile Asp-Pro sequence to enable specific cleavage at the boundary of these domains. Under nonreducing conditions, the amino terminus was released from the receptor body, supporting the absence of covalent association between these domains. Quantitative [(14)C]iodoacetamide incorporation into the isolated amino-terminal domain of the receptor in the absence and presence of chemical reduction established the ratio of free to total Cys residues as 1:7, consistent with three disulfide bonds. Mutagenesis of each of the amino-terminal Cys residues to Ala was tolerated only for Cys(11), suggesting that these bonds linked the conserved Cys residues. This was further supported by treatment of intact cells expressing wild-type or C11A mutant secretin receptor with a cell-impermeant sulfhydryl-reactive reagent. Thus, the functionally important amino terminus of the secretin receptor represents a structurally independent, highly folded, and disulfide-bonded domain, with a pattern that is likely critical and conserved throughout this receptor family. PMID- 11040038 TI - Oxaliplatin-induced damage of cellular DNA. AB - Damage to cellular DNA is believed to determine the antiproliferative properties of platinum (Pt) drugs. This study characterized DNA damage by oxaliplatin, a diaminocyclohexane Pt drug with clinical antitumor activity. Compared with cisplatin, oxaliplatin formed significantly fewer Pt-DNA adducts (e.g., 0.86+/ 0.04 versus 1.36+/- 0.01 adducts/10(6) base pairs/10 microM drug/1 h, respectively, in CEM cells, P<.01). Oxaliplatin was found to induce potentially lethal bifunctional lesions, such as interstrand DNA cross-links (ISC) and DNA protein cross-links (DPC) in CEM cells. As with total adducts, however, oxaliplatin produced fewer (P<.05) bifunctional lesions than did cisplatin: 0.7+/ 0.2 and 1.8+/-0.3 ISC and 0.8+/-0.1 and 1.5+/-0.3 DPC/10(6) base pairs/10 microM drug, respectively, after a 4-h treatment. Extended postincubation (up to 12 h) did not compensate the lower DPC and ISC levels by oxaliplatin. ISC and DPC determinations in isolated CEM nuclei unequivocally verified that oxaliplatin is inherently less able than cisplatin to form these lesions. Reactivation of drug treated plasmids, observed in four cell lines, suggests that oxaliplatin adducts are repaired with similar kinetics as cisplatin adducts. Oxaliplatin, however, was more efficient than cisplatin per equal number of DNA adducts in inhibiting DNA chain elongation ( approximately 7-fold in CEM cells). Despite lower DNA reactivity, oxaliplatin exhibited similar or greater cytotoxicity in several other human tumor cell lines (50% growth inhibition in CEM cells at 1.1/1.2 microM, respectively). The results demonstrate that oxaliplatin-induced DNA lesions, including ISC and DPC, are likely to contribute to the drug's biological properties. However, oxaliplatin requires fewer DNA lesions than does cisplatin to achieve cell growth inhibition. PMID- 11040039 TI - Cyclosaligenyl-2',3'-didehydro-2',3'-dideoxythymidine monophosphate: efficient intracellular delivery of d4TMP. AB - Cyclosaligenyl-2',3'-didehydro-2', 3'-dideoxythymidine-5'-monophosphate (cycloSal d4TMP) is a potent and selective inhibitor of human immunodeficiency virus replication in cell culture and differs from other nucleotide prodrug approaches in that it is designed to selectively deliver the nucleotide 5'-monophosphate by a controlled, chemically induced hydrolysis. Its antiviral efficacy in cell culture is at least as good as, if not superior to, that of d4T. CycloSal-d4TMP was found to lead to the efficient intracellular release of d4TMP in a variety of cell lines, including both wild-type CEM and thymidine kinase-deficient CEM/TK(-) cells. Under similar experimental conditions, exposure of CEM/TK(-) cells to d4T failed to result in significant d4TTP levels. The intracellular conversion of cycloSal-d4TMP proved to be both time and dose dependent. The half-life of d4TTP generated intracellularly from d4T- or cycloSal-d4TMP-treated CEM cells was approximately 3.5 h, and the intracellular ratios of d4TTP/d4TMP in cells exposed to cycloSal-d4TMP gradually increased from 1 to 3.4 upon prolonged incubation. Radiolabeled cycloSal-d4TMP could be separated as its two R(p) and S(p) diastereomers on high-performance liquid chromatography. The R(p) diastereomer of cycloSal-d4TMP was 3- to 7-fold more efficient in releasing d4TMP and generating d4TTP than the S(p) cycloSal-d4TMP diastereomer. This correlated well with the 5 fold more pronounced antiviral activity of the R(p) diastereomer versus the S(p) diastereomer. d4TMP is a poor substrate for the cytosolic 5'(3') deoxyribonucleotidase (V(max)/K(m) for d4TMP: 0.08 of V(max)/K(m) for dTMP) and is only slowly hydrolyzed to d4T. This contributes to the efficient conversion of the prodrug of d4TTP. PMID- 11040040 TI - Characterization of calcium signaling by purinergic receptor-channels expressed in excitable cells. AB - ATP-gated purinergic receptors (P2XRs) are a family of cation-permeable channels that conduct Ca(2+) and facilitate voltage-sensitive Ca(2+) entry in excitable cells. To study Ca(2+) signaling by P2XRs and its dependence on voltage-sensitive Ca(2+) influx, we expressed eight cloned P2XR subtypes individually in gonadotropin-releasing hormone-secreting neurons. In all cases, ATP evoked an inward current and a rise in [Ca(2+)](i). P2XR subtypes differed in the peak amplitude of [Ca(2+)](i) response independently of the level of receptor expression, with the following order: P2X(1)R < P2X(3)R < P2X(4)R < P2X(2b)R < P2X(2a)R < P2X(7)R. During prolonged agonist stimulation, Ca(2+) signals desensitized with different rates: P2X(3)R > P2X(1)R > P2X(2b)R > P2X(4)R >> P2X(2a)R >> P2X(7)R. The pattern of [Ca(2+)](i) response for each P2XR subtype was highly comparable with that of the depolarizing current, but the activation and desensitization rates were faster for the current than for [Ca(2+)](i). The P2X(1)R, P2X(3)R, and P2X(4)R-derived [Ca(2+)](i) signals were predominantly dependent on activation of voltage-sensitive Ca(2+) influx, both voltage sensitive and -insensitive Ca(2+) entry pathways equally contributed to [Ca(2+)](i) responses in P2X(2a)R- and P2X(2b)R-expressing cells, and P2X(7)R operated as a nonselective pore capable of conducting larger amounts of Ca(2+) independently on the status of voltage-gated Ca(2+) channels. Thus, Ca(2+) signaling by homomeric P2XRs expressed in an excitable cell is subtype-specific, which provides an effective mechanism for generating variable [Ca(2+)](i) patterns in response to a common agonist. PMID- 11040041 TI - Evidence that Galpha(q)-coupled receptor-induced interleukin-6 mRNA in vascular smooth muscle cells involves the nuclear factor of activated T cells. AB - The immunosuppressant cyclosporin A inhibits transcription mediated by the nuclear factor of activated T-cells (NFAT), a key regulator of cytokine gene expression in lymphocytes that integrates phospholipase C signaling. NFAT is also expressed in vascular smooth muscle cells, but the genes it regulates there are unknown. Here we show that Galpha(q)-coupled P2Y nucleotide receptor signaling in rat vascular smooth muscle cells increases NFAT-mediated luciferase reporter expression. It also induces interleukin (IL)-6 gene expression but not other cytokine mRNAs including IL-1, IL-2, IL-3, IL-4, IL-10, gamma-interferon, tumor necrosis factor-alpha, or tumor necrosis factor-beta. IL-6 mRNA induction by UTP is more rapid and transient then that caused by IL-1beta stimulation and is partially blocked by cyclosporin A or by expression of a trans-dominant NFAT inhibitor. Expression of recombinant NFATc1 markedly augments IL-6 mRNA induction by these and other agonists, which is partially attributable to NFAT-regulated paracrine mediators. However, trans-dominant NFkappaB inhibitors strongly interfere with IL-6 mRNA induction both by IL-1beta and by UTP, which synergistically evoke IL-6 mRNA expression. These findings suggest that NFAT is among the cofactors involved in NFkappaB-dependent IL-6 gene induction by Ca(2+) mobilizing receptors in vascular smooth muscle cells. PMID- 11040042 TI - Molecular analysis of beta(2)-adrenoceptor coupling to G(s)-, G(i)-, and G(q) proteins. AB - The beta(2)-adrenoceptor (beta(2)AR) couples to the G-protein G(s) to activate adenylyl cyclase. Intriguingly, several studies have demonstrated that the beta(2)AR can also interact with G-proteins of the G(i)- and G(q)-family. To assess the efficiency of beta(2)AR interaction with various G-protein alpha subunits (G(xalpha)), we expressed fusion proteins of the beta(2)AR with the long (G(salphaL)) and short (G(salphaS)) splice variants of G(salpha), the G(i) proteins G(ialpha2) and G(ialpha3), and the G(q)-proteins G(qalpha) and G(16alpha) in Sf9 cells. Fusion proteins provide a rigorous approach for comparing the coupling of a given receptor to G(xalpha) because of the defined 1:1 stoichiometry of receptor and G-protein and the efficient coupling. Here, we show that the beta(2)AR couples to G(s)-, G(i)-, and G(q)-proteins as assessed by ternary complex formation and ligand-regulated guanosine 5'-O-(3 thiotriphosphate) (GTPgammaS) binding. The combined analysis of ternary complex formation, GTPgammaS binding, agonist efficacies, and agonist potencies revealed substantial differences in the interaction of the beta(2)AR with the various classes of G-proteins. Comparison of the coupling of the beta(2)AR and formyl peptide receptor to G(ialpha2) revealed receptor-specific differences in the kinetics of GTPgammaS binding. We also detected highly efficient stimulation of GTPgammaS dissociation from G(salphaL), but not from G(qalpha) and G(16alpha), by a beta(2)AR agonist. Moreover, we show that the 1:1 stoichiometry of receptor to G-protein in fusion proteins reflects the in vivo stoichiometry of receptor/G protein coupling more closely than was previously assumed. Collectively, our data show 1) that the beta(2)AR couples differentially to G(s)-, G(i)-, and G(q) proteins, 2) that there is ligand-specific coupling of the beta(2)AR to G proteins, 3) that receptor-specific G-protein conformational states may exist, and 4) that nucleotide dissociation is an important mechanism for G-protein deactivation. PMID- 11040043 TI - Structure-related inhibition of calmodulin-dependent neuronal nitric-oxide synthase activity by melatonin and synthetic kynurenines. AB - We recently described that melatonin and some kynurenines modulate the N-methyl-D aspartate-dependent excitatory response in rat striatal neurons, an effect that could be related to their inhibition of nNOS. In this report, we studied the effect of melatonin and these kynurenines on nNOS activity in both rat striatal homogenate and purified rat brain nNOS. In homogenates of rat striatum, melatonin inhibits nNOS activity, whereas synthetic kynurenines act in a structure-related manner. Kynurenines carrying an NH(2) group in their benzenic ring (NH(2) kynurenines) inhibit nNOS activity more strongly than melatonin itself. However, kynurenines lacking the NH(2) group or with this group blocked do not affect enzyme activity. Kinetic analysis shows that melatonin and NH(2)-kynurenines behave as noncompetitive inhibitors of nNOS. Using purified rat brain nNOS, we show that the inhibitory effect of melatonin and NH(2)-kynurenines on the enzyme activity diminishes with increasing amounts of calmodulin in the incubation medium. However, changes in other nNOS cofactors such as FAD or H(4)-biopterin, do not modify the drugs' response. These data suggest that calmodulin may be involved in the nNOS inhibition by these compounds. Studies with urea polyacrylamide gel electrophoresis further support an interaction between melatonin and NH(2)-kynurenines, but not kynurenines lacking the NH(2) group, with Ca(2+)-calmodulin yielding Ca(2+)-calmodulin-drug complexes that prevent nNOS activation. The results show that calmodulin is a target involved in the intracellular effects of melatonin and some melatonin-related kynurenines that may account, at least in part, for the neuroprotective properties of these compounds. PMID- 11040044 TI - Fibronectin-mediated hepatocyte shape change reprograms cytochrome P450 2C11 gene expression via an integrin-signaled induction of ribonuclease activity. AB - A major limitation to the use of rat hepatocytes in the study of drug metabolism and toxicity is the rapid loss of CYPs. We demonstrate that the culture of rat hepatocytes results in a rapid loss of liver-specific CYP2C11 mRNA and transcripts encoding the general housekeeping gene copper-zinc superoxide dismutase (CuZnSOD) as well as poly(A(+)) mRNA. These losses are accelerated by fibronectin, which has no effect on the transcription of CYP2C11 and CuZnSOD. However, fibronectin, an extracellular matrix protein involved in cell adhesion and spreading, induces ribonuclease (RNase) activity. Fibronectin also increases hepatocyte diameter and data are presented that cell spreading is involved in the loss of both CYP2C11 and CuZnSOD mRNAs. The use of functional blocking antibodies demonstrates that fibronectin is operating through its alpha(5)beta(1) integrin receptor and genistein, a tyrosine kinase inhibitor, prevents hepatocyte spreading, RNase induction, and CYP2C11 mRNA loss. Collectively, the data indicate that hepatocytes in vitro actively promote the extinction of their phenotype via the autocrine effects of fibronectin rather than the current consensus that they simply lose differentiated function, such as CYP2C11 expression, through the absence of extracellular matrix proteins. The substrate specificity of the ribonuclease induced is also considered. PMID- 11040045 TI - Analysis of the interaction between the HIV-inactivating protein cyanovirin-N and soluble forms of the envelope glycoproteins gp120 and gp41. AB - The novel virucidal protein cyanovirin-N (CV-N) binds with equally high affinity to soluble forms of either H9 cell-produced or recombinant glycosylated HIV-1 gp120 (sgp120) or gp160 (sgp160). Fluorescence polarization studies showed that CV-N is also capable of binding to the glycosylated ectodomain of the HIV envelope protein gp41 (sgp41) (as well as SIV glycoprotein 32), albeit with considerably lower affinity than the sgp120/CV-N interaction. Pretreatment of CV N with either sgp120 or sgp41 abrogated the neutralizing activity of CV-N against intact, infectious HIV-1 virions. Isothermal calorimetry and optical biosensor binding studies showed that CV-N bound to recombinant sgp120 with a K(d) value ranging from 2 to 45 nM and to sgp41 with a K(d) value of 606 nM; furthermore, they indicated an approximate 5:1 stoichiometry for CV-N binding to sgp120 and a 1:1 stoichiometry for CV-N binding to sgp41. Circular dichroism studies additionally illuminated the binding of CV-N with both sgp120 and sgp41, providing the first direct evidence that conformational changes are a consequence of CV-N interactions with both HIV-1 envelope glycoproteins. PMID- 11040046 TI - The amino terminus of Galpha(z) is required for receptor recognition, whereas its alpha4/beta6 loop is essential for inhibition of adenylyl cyclase. AB - G(z) couples to most of the known G(i)-linked receptors and its alpha subunit (Galpha(z)) inhibits adenylyl cyclases as efficiently as Galpha(i) subtypes. A series of chimeric Galpha subunits with different portions of Galpha(z) and Galpha(t1) (a regulator of cGMP phosphodiesterase) were constructed to study the essential structural elements of Galpha(z) that determine receptor coupling and effector interaction. The receptor-mediated functions of the chimeras were assessed in two aspects: 1) stimulation of type 2 adenylyl cyclase through the release of betagamma subunits from the chimeras, and 2) inhibition of isoproterenol-stimulated adenylyl cyclase by the chimeric Galpha subunits. The results suggested that the presence of both termini of Galpha(z) were critical for coupling to delta-opioid receptor, with the N-terminal region being more important. Moreover, a stretch of amino acids (295-319) corresponding to the alpha4/beta6 loop was identified as one of the adenylyl cyclase inhibitory domains of Galpha(z). PMID- 11040047 TI - G(1) phase-dependent expression of bcl-2 mRNA and protein correlates with chemoresistance of human cancer cells. AB - Recent experiments suggest an interconnection between cell proliferation and programmed cell death (apoptosis), although the detailed molecular mechanisms remain unclear. We have hypothesized that expression of some apoptosis regulators is cell cycle-dependent, which in turn influences tumor cell chemosensitivity in a cell cycle-dependent fashion. To test these hypotheses, we synchronized human leukemia Jurkat T, Neo (using aphidicolin), breast cancer MCF-7, normal fibroblast, and simian virus 40-transformed cells (by aphidicolin or serum starvation), and measured levels of several Bcl-2 family proteins. The highest expression of Bcl-2 protein was found in the G(1) phase of all the five cell lines tested. In contrast, levels of Bax protein remained relatively unchanged in four of the cell lines, and levels of Bcl-X(L), Bcl-X(S), and Bak proteins showed little or no cell cycle-dependent changes in Jurkat T cells. Similar to the changes in Bcl-2 protein levels, its mRNA expression was also G(1) phase specific, whereas the level of a Bcl-2 cleavage activity remained constitutive. When treated with an anticancer drug (etoposide or cisplatin) or the kinase inhibitor staurosporin, the cells containing a high G(1) population and a high Bcl-2 protein level were much more resistant to the induced apoptosis than the cells containing a high S phase population and a low Bcl-2 protein level. Constitutive overexpression of Bcl-2 protein in Jurkat T cells completely blocked the S phase-associated sensitivity to these apoptosis stimuli. The cell cycle dependent Bcl-2 protein expression seems to contribute to the regulation of chemosensitivity and apoptotic commitment of human tumor cells. PMID- 11040048 TI - Overexpression of type 7 adenylyl cyclase in the mouse brain enhances acute and chronic actions of morphine. AB - The mechanisms by which morphine-induced analgesia and tolerance and physical dependence on morphine arise have been the subject of intense study, and much work has pointed to the involvement of cAMP-mediated events in the neuroadaptive phenomena leading to morphine tolerance and/or dependence. We overexpressed an opioid receptor-stimulatable form of adenylyl cyclase (type 7) in the central nervous system of mice and demonstrated significant effects of this manipulation on the animals' acute response to morphine, the development of morphine tolerance, and development of sensitization to morphine. Measurements of the acute analgesic response to morphine demonstrated that the ED(50) values for the transgenic mice were significantly lower than the ED(50) values determined for the "wild-type" animals. During chronic treatment with morphine, the transgenic mice developed tolerance more rapidly than the wild-type mice, and transgenic animals of the C57BL/6xSJL background showed a larger sensitization to morphine's effects on locomotor activity than did wild-type mice of the same background. These results indicated that cAMP-generating systems may simultaneously modulate the development of tolerance and sensitization. Interestingly, the signs of physical dependence on morphine in the transgenic mice did not differ from those in their wild-type litter mates, indicating that separate mechanisms may modulate opiate tolerance and opiate dependence. PMID- 11040049 TI - The essential role of phosphatidylinositol 3-kinase and of p38 mitogen-activated protein kinase activation in the antioxidant response element-mediated rGSTA2 induction by decreased glutathione in H4IIE hepatoma cells. AB - The protective adaptive response to electrophiles and reactive oxygen species is mediated by the enhanced expression of the phase II detoxifying genes through antioxidant response elements (AREs). The current study was designed to identify the signaling pathways responsible for the expression of rGSTA2 in response to cellular oxidative stress and to establish the molecular mechanistic basis. Deprivation of cystine and methionine caused oxidative stress in H4IIE hepatoma cells as evidenced by a marked decrease in the reduced glutathione (first order rate constant = 0.056 h(-1); t(1/2) = 12.6 h) and an increase in pro-oxidant production. Electrophoretic mobility shift assay revealed that the ARE complex, consisting of Nrf-1/2 and Maf proteins, was activated 12 to 48 h after sulfur amino acid deprivation (SAAD). The rGSTA2 mRNA level was elevated by SAAD beginning at 24 h, whereas the rGSTA2 subunit was maximally induced at 48 h. Nuclear ARE activation and rGSTA2 mRNA increase were both completely inhibited by wortmannin or LY294002, the phosphatidylinositol 3-kinase (PI3-kinase) inhibitors. The p38 mitogen-activated protein (MAP) kinase was activated at 0.5 to 3 h after SAAD, followed by sustained diminished activation up to 12 h. Inhibition of p38 MAP kinase by SB203580 prevented the ARE-mediated rGSTA2 induction. The activation of p38 MAP kinase, however, failed to be inhibited by wortmannin or LY294002, showing that PI3-kinase is not involved in the activation of p38 MAP kinase. Data showed that PI3-kinase plays an essential role in the ARE mediated rGSTA2 induction by oxidative stress after SAAD, which activates the p38 MAP kinase and leads to rGSTA2 induction. PMID- 11040050 TI - The inhibitory potency and selectivity of arginine substrate site nitric-oxide synthase inhibitors is solely determined by their affinity toward the different isoenzymes. AB - We have investigated various nitric oxide (NO) synthase inhibitors for their affinity and selectivity toward the three human isoenzymes in radioligand binding experiments. Therefore, we developed the new radioligand [(3)H]2-amino-4-picoline to measure binding of these compounds to the three human NO synthase (NOS) isoenzymes. Aminopicoline is a potent and nonselective inhibitor of all three isoforms. [(3)H]2-amino-4-picoline bound saturably and with high affinity to human NOSs. Affinity constants (K(D) values) of 59, 111, and 136 nM were obtained for the inducible, neuronal, and endothelial NOS isoforms (iNOS, nNOS, eNOS). Binding of [(3)H]2-amino-4-picoline was competitive with the substrate arginine. From all the inhibitors tested, AMT (2-amino-5, 6-dihydro-6-methyl-4H-1,3 thiazine hydrochloride) showed the highest affinity and no selectivity. L-NIL [L N(6)-(1-Iminoethyl)lysine hydrochloride] and aminoguanidine were moderately iNOS selective while L-NA (N(G)-nitro-L-arginine) and L-NAME (N(G)-nitro-L-arginine methyl ester hydrochloride) showed selectivity toward the constitutive isoforms. High iNOS versus eNOS selectivity was found for 1400W, whereas several isothiourea derivatives and 1400W displayed moderate n- versus eNOS selectivity. To relate the affinity of these compounds to their inhibitory potency, we measured the inhibitory potency under almost identical conditions using a new microtiter plate assay. The inhibitory potency of selective and nonselective NOS inhibitors was almost exactly mirrored by their affinity toward the different isoenzymes. Highly significant correlations were obtained between the potency of enzyme inhibition and the inhibition of [(3)H]2-amino-4-picoline binding for all three isoenzymes. These data show that the potency and selectivity of NOS inhibitors are solely determined by their affinity toward the different isoforms. Furthermore, these data identify the new radioligand [(3)H]2-amino-4-picoline as a very useful radiolabel for the investigation of the substrate binding site of all three isoforms. PMID- 11040051 TI - Creation of a selective antagonist and agonist of the rat VPAC(1) receptor using a combinatorial approach with vasoactive intestinal peptide 6-23 as template. AB - We have used combinatorial chemistry with amino acid mixtures (X) at positions 6 to 23 in vasoactive intestinal peptide (VIP) to optimize binding affinity and selectivity to the rat VPAC(1) receptor. The most efficient amino acid replacement was a substitution of alanine at position 18 to diphenylalanine (Dip), increasing the displacement efficiency of (125)I-VIP by 370-fold. The [Dip(18)]VIP(6-23) was subsequently used to find a second replacement, employing the same approach. Tyrosine at position 9 was selected and the resulting [Tyr(9),Dip(18)]VIP(6-23) analog has a K(i) value of 90 nM. This analog was unable to stimulate cAMP production at 10(-6) M but was able to inhibit VIP induced cAMP stimulation (K(b) = 79 nM). The K(i) values of [Tyr(9),Dip(18)]VIP(6 23) using the rat VPAC(2) and PAC(1) receptors were 3,000 nM and >10,000 nM, respectively. Thus, [Tyr(9),Dip(18)]VIP(6-23) is a selective VPAC(1) receptor antagonist. The C-terminally extended form, [Tyr(9),Dip(18)]VIP(6-28), displays improved antagonistic properties having a K(i) and K(b) values of 18 nM and 16 nM, respectively. On the contrary, the fully extended form, [Tyr(9),Dip(18)]VIP(1 28), was a potent agonist with improved binding affinity (K(i) = 0.11 nM) and ability to stimulate cAMP (EC(50) = 0.23 nM) compared with VIP (K(i) = 1.7 nM, EC(50) = 1.12 nM). Furthermore, the specificity of this agonist to the VPAC(1) receptor was high, the K(i) values for the VPAC(2) and PAC(1) receptors were 53 nM and 3,100 nM, respectively. Seven other analogs with the [Tyr(9),Dip(18)] replacement combined with previously published VIP modifications have been synthesized and described in this work. PMID- 11040052 TI - Inverse agonism and constitutive activity as functional correlates of serotonin h5-HT(1B) receptor/G-protein stoichiometry. AB - This study evaluated the influence of receptor/G-protein (R:G) stoichiometry on constitutive activity and the efficacy of agonists, partial agonists, and inverse agonists at human (h) 5-hydroxytryphamine 1B (5-HT(1B)) receptors. Two Chinese hamster ovary cell lines were used; they expressed 8.5 versus 0.4 pmol h5-HT(1B) receptors/mg (determined by [(3)H]GR125,743 saturation analysis) and 3.0 versus 1.5 pmol receptor-activated G-proteins/mg [determined by guanosine-5'-O-(3 [(35)S]thio)-triphosphate ([(35)S]GTPgammaS) isotopic dilution], respectively. Thus, they displayed R:G ratios of approximately 3.0 (RGhigh) and approximately 0.3 (RGlow), respectively. In competition-binding experiments, the agonists, 5-HT and sumatriptan, displayed fewer high-affinity (HA)-binding sites and the partial agonists, BMS181, 101 and L775,606, displayed decreased affinity in RGhigh versus RGlow membranes. In contrast, the inverse agonists, SB224,289 and, to a lesser extent, methiothepin, showed increased affinity. In G-protein activation experiments, both basal and 5-HT-activated [(35)S]GTPgammaS binding were higher in RGhigh than in RGlow membranes. Constitutive activity (determined by inhibition of basal [(35)S]GTPgammaS binding with GTPgammaS in the absence of receptor ligands) was more pronounced in RGhigh versus RGlow membranes, as revealed by the >5-fold greater proportion of HA sites. Correspondingly, the negative efficacy of inverse agonists was strikingly augmented, inasmuch as they suppressed approximately two-thirds of HA [(35)S]GTPgammaS binding in RGhigh membranes, but only approximately one-third in RGlow membranes. Furthermore, the efficacy of partial agonists was greater at RGhigh versus RGlow membranes, as estimated by their ability to enhance [(35)S]GTPgammaS binding. In conclusion, an increase in R:G ratios at h5-HT(1B) receptors was associated with an increase in relative efficacy of partial agonists and, most notably, an increase in both constitutive G-protein activation and negative efficacy of inverse agonists. PMID- 11040053 TI - Identification of G protein-coupled receptor kinase 2 phosphorylation sites responsible for agonist-stimulated delta-opioid receptor phosphorylation. AB - Agonist-induced receptor phosphorylation is an initial step in opioid receptor desensitization, a molecular mechanism of opioid tolerance and dependence. Our previous research suggested that agonist-induced delta-opioid receptor (DOR) phosphorylation occurs at the receptor carboxyl terminal domain. The current study was carried out to identify the site of DOR phosphorylation during agonist stimulation and the kinases catalyzing this reaction. Truncation (Delta15) or substitutions (T358A, T361A, and S363G single or triple mutants) at the DOR cytoplasmic tail caused 80 to 100% loss of opioid-stimulated receptor phosphorylation, indicating that T358, T361, and S363 all contribute and are cooperatively involved in agonist-stimulated DOR phosphorylation. Coexpression of GRK2 strongly enhanced agonist-stimulated phosphorylation of the wild-type DOR (WT), but Delta15 or mutant DOR (T358A/T361A/S363G) failed to show any detectable phosphorylation under these conditions. These results demonstrate that T358, T361, and S363 are required for agonist-induced and GRK-mediated receptor phosphorylation. Agonist-induced receptor phosphorylation was severely impaired by substitution of either T358 or S363 with aspartic acid residue, but phosphorylation of the T361D mutant was comparable with that of WT. In the presence of exogenously expressed GRK2, phosphorylation levels of T358D and S363D mutants were approximately half of that of WT, whereas significant phosphorylation of the T358/S363 double-point mutant was not detected. These results indicate that both T358 and S363 residues at the DOR carboxyl terminus are capable of serving cooperatively as phosphate acceptor sites of GRK2 in vivo. Taken together, we have demonstrated that agonist-induced opioid receptor phosphorylation occurs exclusively at two phosphate acceptor sites (T358 and S363) of GRK2 at the DOR carboxyl terminus. These results represent the identification of the GRK phosphorylation site on an opioid receptor for the first time and demonstrate that GRK is the prominent kinase responsible for agonist-induced opioid receptor phosphorylation in vivo. PMID- 11040054 TI - Regulation of cell proliferation, gene expression, production of cytokines, and cell cycle progression in primary human T lymphocytes by piperlactam S isolated from Piper kadsura. AB - Effects of piperlactam S (C(17)H(13)NO(4); mol. wt. 295) isolated from Piper kadsura on phytohemagglutinin (PHA) stimulated cell proliferation were studied in primary culture of human T cells. The results showed that piperlactam S suppressed T cell proliferation at about 0 to 12 h after stimulation with PHA. Synthesis of total cellular proteins and RNA in activated cell cultures was also suppressed. The inhibitory action of piperlactam S was not through direct cytotoxicity. Cell cycle analysis indicated that piperlactam S arrested the cell cycle progression of activated T cells from the G(1) transition to the S phase. In an attempt to further localize the point in the cell cycle at which arrest occurred, a set of key regulatory events leading to the G(1)/S boundary, including gene expression of cytokines and c-Fos protein synthesis, was examined. Piperlactam S suppressed, in activated T lymphocytes, the production and mRNA expression of cytokines such as interleukin-2 (IL-2), IL-4, and interferon-gamma in a dose-dependent manner. In addition, Western blot analysis indicated that c Fos protein expressed in activated T lymphocytes was decreased by piperlactam S. Results of kinetic study indicated that inhibitory effects of piperlactam S on IL 2 mRNA expressed in T cells might be related to blocking c-Fos protein synthesis. Thus, the suppressant effects of piperlactam S on proliferation of T cells activated by PHA seemed to be mediated, at least in part, through inhibition of early transcripts of T cells, especially those of important cytokines, IL-2, IL 4, and arresting cell cycle progression in the cells. PMID- 11040055 TI - Structure activity relationship of carboxylic ester antagonists of the vitamin D(3) receptor. AB - A 25-carboxylic ester analog of 1alpha,25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)], ZK159222 (compound 1), was recently described as a novel type of antagonist of 1alpha,25(OH)(2)D(3) signaling. In this study five derivatives of compound 1 (compounds 2-6) were selected because of their sensitivity in facilitating complex formation between the 1alpha,25(OH)(2)D(3) receptor (VDR) and the retinoid X receptor on a 1alpha,25(OH)(2)D(3) response element that was comparable to that of the natural hormone (0.2-0.9 nM). Most derivatives of compound 1 reacted as typical agonists, because they were able to promote ligand-dependent interaction of the VDR with the coactivator TIF2, stabilized the VDR preferentially in its agonistic conformation c1(LPD), and stimulated VDR-dependent gene activity with a potency similar to 1alpha,25(OH)(2)D(3). In contrast, only compound 2 showed the antagonistic profile of compound 1, which includes the incompetence to induce a VDR-TIF2 contact, the stabilization of the antagonistic conformation c2(LPD), and only a very weak and insensitive functional activity. Accordingly, only compounds 1 and 2, but not compounds 3 to 6, showed prominent antagonistic effects in cellular systems. The comparison of the structures of the compounds indicates that the essential requirements for an antagonistic function are a cyclopropyl ring at carbon 25, a hydroxy group at carbon 24, and at least a butylester. Interestingly, compound 2 was an approximately 3 times more sensitive antagonist than compound 1 and even displayed a lower residual agonistic activity. In conclusion, only a very limited number of structural variations of compound 1 are possible to keep its antagonistic profile, but the tools presented here for their in vitro evaluation allow an accurate prediction of the effects and are suited to screening for even more potent 1alpha, 25(OH)(2)D(3) antagonists. PMID- 11040056 TI - Influence of receptor number on functional responses elicited by agonists acting at the human adenosine A(1) receptor: evidence for signaling pathway-dependent changes in agonist potency and relative intrinsic activity. AB - Activation of A(1) adenosine receptors leads to the inhibition of cAMP accumulation and the stimulation of inositol phosphate accumulation via pertussis toxin-sensitive G-proteins. In this study we have investigated the signaling of the A(1) adenosine receptor in Chinese hamster ovary (CHO) cells, when expressed at approximately 203 fmol/mg (CHOA1L) and at approximately 3350 fmol/mg (CHOA1H). In CHOA1L cells, the agonists N(6)-cyclopentyladenosine (CPA), (R)-N(6)-(2 phenylisopropyl)adenosine, and 5'-(N-ethylcarboxamido)adenosine (NECA) inhibited cAMP production in a concentration-dependent manner. After pertussis toxin treatment, the agonist NECA produced a stimulation of cAMP production, whereas CPA and (R)-N(6)-(2-phenylisopropyl)adenosine were ineffective. In CHOAIH cells, however, all three agonists produced both an inhibition of adenylyl cyclase and a pertussis toxin-insensitive stimulation of adenylyl cyclase. All three agonists were more potent at inhibiting adenylyl cyclase in CHOA1H cells than in CHOA1L cells. In contrast, A(1) agonists (and particularly NECA) were less potent at stimulating inositol phosphate accumulation in CHOA1H cells than in CHOA1L cells. After pertussis toxin treatment, agonist-stimulated inositol phosphate accumulation was reduced in CHOA1H cells and abolished in CHOA1L cells. The relative intrinsic activity of NECA in stimulating inositol phosphate accumulation, compared to CPA (100%), was much greater in the presence of pertussis toxin (289.6%) than in the absence of pertussis toxin (155.2%). These data suggest that A(1) adenosine receptors can couple to both pertussis toxin sensitive and -insensitive G-proteins in an expression level-dependent manner. These data also suggest that the ability of this receptor to activate different G proteins is dependent on the agonist present. PMID- 11040057 TI - Analysis of the pharmacological and molecular heterogeneity of I(2)-imidazoline binding proteins using monoamine oxidase-deficient mouse models. AB - The I(2) subgroup of imidazoline-binding sites was identified as monoamine oxidases (MAOs), but it is unclear whether there are I(2)-binding sites located on proteins distinct from MAOs. To address this issue, we characterized I(2) binding proteins in liver and brain of wild-type and MAO A- and MAO B-deficient mice. I(2)-binding sites were identified using [(3)H]idazoxan and the photoaffinity adduct 2-[3-azido-4-[(125)I]iodophenoxyl]methylimidazoline ([(125)I]AZIPI). [(3)H]Idazoxan labeled binding sites with ligand recognition properties typical of I(2) sites in both brain and liver of wild-type mice. High affinity, specific [(3)H]idazoxan binding were not altered in MAO A knockout (KO) mice. In contrast, [(3)H]idazoxan binding was completely abolished in both liver and brain of MAO B KO mice. In wild-type mice, [(125)I]AZIPI photolabeled three proteins with apparent molecular masses of approximately 28 (liver), approximately 61 (brain), and approximately 55 kDa (liver and brain). The photolabeling of each protein was blocked by the imidazoline cirazoline (10 microM). Photolabeling of the approximately 61- and approximately 55-kDa proteins was not observed in MAO A and B KO mice, respectively. In contrast, photolabeling of the liver approximately 28-kDa protein was still observed in MAO-deficient mice, indicating that this protein is unrelated to MAOs. These data indicate that I(2) imidazoline-binding sites identified by [(3)H]idazoxan reside solely on MAO B. The binding sites on MAO A and the liver approximately 28-kDa protein may represent additional subtypes of the family of the imidazoline-binding sites. PMID- 11040058 TI - Agonist binding and function at the human alpha(2A)-adrenoceptor: allosteric modulation by amilorides. AB - It has been found previously that amilorides act via an allosteric site on the alpha(2A)-adrenergic receptor to strongly inhibit antagonist binding. In this study, allosteric modulation of agonist binding and function at the alpha(2A) adrenergic receptor was explored. The dissociation rate of the agonist [(3)H]UK14304 from alpha(2A)-receptors was decreased by the amilorides in a concentration-dependent manner. This contrasts with the increases in (3)H antagonist dissociation rate found previously. The agonist-amiloride analog interaction data could be fitted to equations derived from the ternary complex allosteric model. The calculated log affinities of the amilorides at the [(3)H]UK14304-occupied receptor increased with the size of the 5-N-alkyl side chain and ranged from 2.4 for amiloride to 4.2 for 5-(N,N-hexamethylene) amiloride. The calculated negative cooperativities cover a narrow range, in sharp contrast to the broad range found for antagonist-amiloride analog interactions. The effects of the amilorides on the agonist actions of UK14304, epinephrine, and norepinephrine were explored using a [(35)S]GTPgammaS functional assay, and the parameters calculated for the cooperativities and affinities of the UK14304 amiloride analog interactions, using the equation derived from the ternary complex allosteric model, were in good agreement with those derived from the kinetic studies. Therefore both the binding and functional data provide further support for the existence of a well defined allosteric site on the human alpha(2A)-adrenergic receptor. The binding mode of the amilorides at the agonist occupied and antagonist-occupied receptor differs markedly but, within each group, the structure of either the agonist or the antagonist examined has only a slight effect on the allosteric interactions. PMID- 11040059 TI - Polyanionic (i.e., polysulfonate) dendrimers can inhibit the replication of human immunodeficiency virus by interfering with both virus adsorption and later steps (reverse transcriptase/integrase) in the virus replicative cycle. AB - Polyanionic dendrimers were synthesized and evaluated for their antiviral effects. Phenyldicarboxylic acid (BRI6195) and naphthyldisulfonic acid (BRI2923) dendrimers were found to inhibit the replication of human immunodeficiency virus type 1 (HIV-1; strain III(B)) in MT-4 cells at a EC(50) of 0.1 and 0.3 microg/ml, respectively. The dendrimers were not toxic to MT-4 cells up to the highest concentrations tested (250 microg/ml). These compounds were also effective against various other HIV-1 strains, including clinical isolates, HIV-2 strains, simian immunodeficiency virus (SIV, strain MAC(251)), and HIV-1 strains that were resistant to reverse transcriptase inhibitors. HIV strains containing mutations in the envelope glycoprotein gp120 (engendering resistance to known adsorption inhibitors) displayed reduced sensitivity to the dendrimers. The compounds inhibited the binding of wild-type virus and recombinant virus (containing wild type gp120) to MT-4 cells at concentrations comparable to those that inhibited the replication of HIV-1(III(B)) in these cells. Cellular uptake studies indicated that BRI2923, but not BRI6195, permeates into MT-4 and CEM cells. Accordingly, the naphtyldisulfonic acid dendrimer (BRI2923) proved able to inhibit later steps of the replication cycle of HIV, i.e., reverse transcriptase and integrase. NL4.3 strains resistant to BRI2923 were selected after passage of the virus in the presence of increasing concentrations of BRI2923. The virus mutants showed 15-fold reduced sensitivity to BRI2923 and cross-resistance to known adsorption inhibitors. However, these virus mutants were not cross resistant to reverse transcriptase inhibitors or protease inhibitors. We identified several mutations in the envelope glycoprotein gp120 gene (i.e., V2, V3, and C3, V4, and C4 regions) of the BRI2923-resistant NL4.3 strains that were not present in the wild-type NL4.3 strain, whereas no mutations were found in the reverse transcriptase or integrase genes. PMID- 11040060 TI - Metabolism and mode of inhibition of varicella-zoster virus by L-beta-5 bromovinyl-(2-hydroxymethyl)-(1,3-dioxolanyl)uracil is dependent on viral thymidine kinase. AB - A nonnaturally occurring L-configuration nucleoside analog, L-beta-5-bromovinyl (2-hydroxymethyl)-1,3-(dioxolanyl)uracil (L-BVOddU) selectively inhibited varicella-zoster virus growth in human embryonic lung (HEL) 299 cell culture with an EC(50) of 0.055 microM, whereas no inhibition of CEM and HEL 299 cell growth or mitochondrial DNA synthesis was observed at concentrations up to 200 microM. L BVOddU was phosphorylated by viral thymidine kinase but not by human cytosolic thymidine kinase, and the antiviral activity of this compound is dependent on the viral thymidine kinase. Unlike other D-configuration bromovinyl deoxyuridine analogs, such as E-5-(2-bromovinyl)-2'-deoxyuridine and 1-beta-arabinofuranosyl-E 5-(2-bromovinyl)uracil, this compound was metabolized only to its monophosphate metabolite. The di- or triphosphate metabolites were not detected. This suggested that the inhibitory mechanism may be unique and different from other anti herpesvirus nucleoside analogs. PMID- 11040062 TI - Benzodiazepines induce a conformational change in the region of the gamma aminobutyric acid type A receptor alpha(1)-subunit M3 membrane-spanning segment. AB - Benzodiazepine binding to gamma-aminobutyric acid type A (GABA(A)) receptors allosterically modulates GABA binding and increases the currents induced by submaximal GABA concentrations. Benzodiazepines induce conformational changes in the GABA-binding site in the extracellular domain, but it is uncertain whether these conformational changes extend into the membrane-spanning domain where the channel gate is located. Alone, benzodiazepines do not open the channel. We used the substituted-cysteine-accessibility method to investigate diazepam-induced conformational changes in the region of the alpha(1)-subunit M3 membrane-spanning segment. In the absence of diazepam or GABA, pCMBS(-) did not react at a measurable rate with cysteine-substitution mutants between alpha(1)Phe296 and alpha(1)Glu303. In the presence of 100 nM diazepam, pCMBS(-) reacted with alpha(1)F296C, alpha(1)F298C, and alpha(1)L301C but not with the other cysteine mutants between alpha(1)Phe296 and alpha(1)Glu303. These three mutants are a subset of the five residues that we previously showed reacted with pCMBS(-) applied in the presence of GABA. The pCMBS(-) reaction rates with these three cysteine mutants were similar in the presence of diazepam and GABA. Thus, diazepam, which binds to the extracellular domain, induces a conformational change in the membrane-spanning domain that is similar to a portion of the change induced by GABA. Because diazepam does not open the channel, these results provide structural evidence that the diazepam-bound state represents an intermediate conformation distinct from the open and resting/closed states of the receptor. The diazepam-induced conformational change in the M3 segment vicinity may be related to the mechanism of allosteric potentiation. PMID- 11040061 TI - Pharmacological blockade of ERG K(+) channels and Ca(2+) influx through store operated channels exerts opposite effects on intracellular Ca(2+) oscillations in pituitary GH(3) cells. AB - In the present study, the effects on intracellular calcium concentration ([Ca(2+)](i)) oscillations of the blockade of ether-a-go-go-related gene (ERG) K(+) channels and of Ca(2+) influx through store-operated channels (SOC) activated by [Ca(2+)](i) store depletion have been studied in GH(3) cells by means of a combination of single-cell fura-2 microfluorimetry and whole-cell mode of the patch-clamp technique. Nanomolar concentrations (1-30 nM) of the piperidinic second-generation antihistamines terfenadine and astemizole and of the class III antiarrhythmic methanesulfonanilide dofetilide, by blocking ERG K(+) channels, increased the frequency and the amplitude of [Ca(2+)](i) oscillations in resting oscillating GH(3) cells. These compounds also induced the appearance of an oscillatory pattern of [Ca(2+)](i) in a subpopulation of nonoscillating GH(3) cells. The effects of ERG K(+) channel blockade on [Ca(2+)](i) oscillations appeared to be due to the activation of L-type Ca(2+) channels, because they were prevented by 300 nM nimodipine. By contrast, the piperazinic second-generation antihistamine cetirizine (0.01-30 microM), which served as a negative control, failed to affect ERG K(+) channels and did not interfere with [Ca(2+)](i) oscillations in GH(3) cells. Interestingly, micromolar concentrations of terfenadine and astemizole (0.3-30 microM), but not of dofetilide (10-100 microM), produced an inhibition of the spontaneous oscillatory pattern of [Ca(2+)](i) changes. This effect was possibly related to an inhibition of SOC, because these compounds inhibited the increase of [Ca(2+)](i) achieved by extracellular calcium reintroduction after intracellular calcium store depletion with the sarcoplasmic or endoplasmic reticulum calcium ATPase pump inhibitor thapsigargin (10 microM) in an extracellular calcium-free medium. The same inhibitory effect on [Ca(2+)](i) oscillations and SOC was observed with the first generation antihistamine hydroxyzine (1-30 microM), the more hydrophobic metabolic precursor of cetirizine. Collectively, the results of the present study obtained with compounds that interfere in a different concentration range with ERG K(+) channels or SOC suggest that 1) ERG K(+) channels play a relevant role in controlling the oscillatory pattern of [Ca(2+)](i) in resting GH(3) cells and 2) the inhibition of SOC might induce an opposite effect, i.e., an inhibition of [Ca(2+)](i) oscillations. PMID- 11040063 TI - Sodium dodecyl sulfate-stable complexes of echistatin and RGD-dependent integrins: a novel approach to study integrins. AB - This study shows that disintegrins, echistatin as a model, can be used as a radiolabeled probe to simultaneously detect the presence of individual RGD dependent integrins on cardiac fibroblasts. Binding of (125)I-echistatin to fibroblasts was proportional to cell number, time dependent, reversible, saturable, specific, and membrane bound. SDS-polyacrylamide gel electrophoresis and autoradiograms revealed that (125)I-echistatin was associated with three radioactive protein bands of 180, 210, and 220 kDa that were identified by RGD affinity chromatography, immunoblotting, and immunoneutralization as alpha(v)beta(3), alpha(3)beta(1)/alpha(5)beta(1)/alpha(v)beta(1), and alpha(8)beta(1) heterodimeric integrins, respectively. These results suggest that echistatin binds to RGD-dependent integrins, forming SDS-stable complexes in the absence of chemical cross-linkers, reducing conditions and heating. As assessed by radioligand-binding filtration, disintegrins displayed binding characteristics with an IC(50) ranging from 0.044 to 1.1 nM, but with slope factors lower than 1, indicating the presence of several binding sites. Resolved by SDS-polyacrylamide gel electrophoresis to reveal echistatin-integrin complexes, disintegrins and RGD peptides displayed different binding affinities to individual RGD-dependent integrins present on cardiac fibroblasts. Elegantin and flavostatin demonstrated the highest affinity toward integrins, whereas flavoridin and acPenRGDC had a greater specificity toward alpha(v)beta(3)-integrin. In summary, echistatin forms SDS-stable complexes with RGD-dependent integrins. This model offers a novel way to visualize RGD-dependent integrins, to investigate their activation state, and to determine the integrin specificity of RGD peptides. PMID- 11040064 TI - Establishment of an isogenic human colon tumor model for NQO1 gene expression: application to investigate the role of DT-diaphorase in bioreductive drug activation in vitro and in vivo. AB - Many tumors overexpress the NQO1 gene, which encodes DT-diaphorase (NADPH:quinone oxidoreductase; EC 1.6.99.2). This obligate two-electron reductase deactivates toxins and activates bioreductive anticancer drugs. We describe the establishment of an isogenic human tumor cell model for DT-diaphorase expression. An expression vector was used in which the human elongation factor 1alpha promoter produces a bicistronic message containing the genes for human NQO1 and puromycin resistance. This was transfected into the human colon BE tumor line, which has a disabling point mutation in NQO1. Two clones, BE2 and BE5, were selected that were shown by immunoblotting and enzyme activity to stably express high levels of DT diaphorase. Drug response was determined using 96-h exposures compared with the BE vector control. Functional validation of the isogenic model was provided by the much greater sensitivity of the NQO1-transfected cells to the known DT diaphorase substrates and bioreductive agents streptonigrin (113- to 132-fold) and indoloquinone EO9 (17- to 25-fold) and the inhibition of this potentiation by the DT-diaphorase inhibitor dicoumarol. A lower degree of potentiation was seen with the clinically used agent mitomycin C (6- to 7-fold) and the EO9 analogs, EO7 and EO2, that are poorer substrates for DT-diaphorase (5- to 8-fold and 2- to 3-fold potentiation, respectively), and there was no potentiation or protection with menadione and tirapazamine. Exposure time-dependent potentiation was seen with the diaziquone analogs methyl-diaziquone and RH1 [2, 5-diaziridinyl-3 (hydroxymethyl)-6-methyl-1,4-benzoquinone], the latter being an agent in preclinical development. In contrast to the in vitro potentiation, there was no difference in the response to mitomycin C when BE2 and BE vector control were treated as tumor xenografts in vivo. This isogenic model should be valuable for mechanistic studies and bioreductive drug development. PMID- 11040065 TI - Homologous and heterologous phosphorylation of the AT(2) angiotensin receptor by protein kinase C. AB - The angiotensin AT(2) receptor is an atypical seven transmembrane domain receptor that is coupled to activation of tyrosine phosphatase and inhibition of MAP kinase, and does not undergo agonist-induced internalization. An investigation of the occurrence and nature of AT(2) receptor phosphorylation revealed that phorbol ester-induced activation of protein kinase C (PKC) in HA-AT(2) receptor expressing COS-7 cells caused rapid and specific phosphorylation of a single residue (Ser(354)) located in the cytoplasmic tail of the receptor. Agonist activation of AT(2) receptors by angiotensin II (Ang II) also caused rapid PKC dependent phosphorylation of Ser(354) that was prevented by the AT(2) antagonist, PD123177, and by inhibitors of PKC. In cells coexpressing AT(1) and AT(2) receptors, Ang II-induced phosphorylation of the AT(2) receptor was reduced by either PD123177 or the AT(1) receptor antagonist, DuP753, and was abolished by treatment with both antagonists or with PKC inhibitors. These findings indicate that the AT(2) receptor is rapidly phosphorylated via PKC during homologous activation by Ang II, and also undergoes heterologous PKC-dependent phosphorylation during activation of the AT(1) receptor. The latter process may regulate the counteracting effects of AT(2) receptors on growth responses to AT(1) receptor activation. PMID- 11040066 TI - Localization of the sites mediating desensitization of the beta(2)-adrenergic receptor by the GRK pathway. AB - The human beta(2)-adrenergic receptor (betaAR) is rapidly desensitized in response to saturating concentrations of agonist by G protein-coupled receptor kinases (GRKs) and cAMP-dependent protein kinase A (PKA) phosphorylation of the betaAR, followed by beta-arrestin binding and receptor internalization. betaAR sites phosphorylated by GRK in vivo have not yet been identified. In this study, we examined the role of the carboxyl terminal serines, 355, 356, and 364, in the GRK-mediated desensitization of the betaAR. Substitution mutants of these serine residues were constructed in which either all three (S355,356,364A), two (S355,356A and S356, 364A), or one of the serines (S356A and S364A) were modified. These mutants were constructed in a betaAR in which the serines of the PKA consensus site were substituted with alanines (designated PKA(-)) to eliminate any PKA contribution to desensitization, and they were stably transfected into human embryonic kidney 293 cells. Treatment of the PKA(-) mutant with 10 microM epinephrine for 5 min caused a 3. 5-fold increase in the EC(50) value and a 42% decrease in the V(max) value for epinephrine stimulation of adenylyl cyclase. Substitution of all three serines completely inhibited the epinephrine-induced shift in the EC(50). Both double mutants, S355,356A and S356,364A, showed a nearly complete loss of the EC(50) shift, whereas the single substitutions, S356A and S364A, caused only a slight decrease in desensitization. None of the mutations altered the epinephrine-induced decrease in V(max,) which seems to be downstream of the receptor. The triple mutation caused a 45% decrease in epinephrine-induced internalization and a 90 to 95% reduction in phosphorylation of the betaAR relative to the PKA(-) (1.9+/- 0.2- and 16.6+/-3.8 fold phosphorylation over basal, respectively). The double mutants caused an intermediate reduction in internalization (20-21%) and phosphorylation (43-52%). None of the serine mutations altered the rate of betaAR recycling. Our data demonstrate that the cluster of serines within the 355 to 364 betaAR domain confer the rapid, GRK-mediated, receptor-level desensitization of the betaAR. PMID- 11040067 TI - Has GAD a central role in type 1 diabetes? PMID- 11040068 TI - Does GAD have a unique role in triggering IDDM? PMID- 11040069 TI - It's insulin. PMID- 11040070 TI - Patients with bullous pemphigoid and linear IgA disease show a dual IgA and IgG autoimmune response to BP180. AB - Bullous pemphigoid (BP) and linear IgA disease (LAD) are autoimmune subepidermal blistering skin diseases associated with autoantibodies against the transmembrane hemidesmosomal protein BP180/type XVII collagen. It has been demonstrated previously that BP is characterized predominantly by IgG autoantibodies, while autoantibodies in LAD mainly belong to the IgA isotype. The aim of the present study was to investigate the hypothesis that there is a significant overlap in the autoantibody isotype profiles associated with these two diseases. Several new recombinant forms of BP180 were generated in the baculovirus expression system, including the full-length protein. IgG autoantibodies to BP 180 were detectable in 39 of 40 (98%) of BP sera; interestingly, 88% of BP sera also contained IgA anti-BP180 autoantibodies. Similarly, anti-BP180 reactivity in LAD sera (n=22) was also attributed to both an IgA (68%) and an IgG (76%) autoantibody response. IgA and IgG autoantibodies to the intracellular portion of BP180 were found in 14% and 28% of BP sera, respectively, and in 8% of LAD sera (same percentage for both isotypes). Our findings clearly demonstrate that both BP and LAD patients have a dual IgA and IgG autoimmune response to BP180 which is directed not only to the ectodomain, but also to the intracellular portion of this protein. PMID- 11040071 TI - Grafting of fibroblasts isolated from the synovial membrane of rheumatoid arthritis (RA) patients induces chronic arthritis in SCID mice-A novel model for studying the arthritogenic role of RA fibroblasts in vivo. AB - The objective of this study was to verify whether isolated rheumatoid arthritis (RA) synovial fibroblasts induce chronic arthritis in SCID mice, in analogy to whole tissue pieces. Fibroblasts were isolated from the synovial membrane of four RA patients (or controls) by out-growth and repeated-passage culture. Following flow-cytometry characterization, 2x10(6)cells were transferred into the left knee joint of SCID mice. The development of arthritis was assessed by joint swelling and histological changes. Human and murine cytokines were measured in vitro in co cultures (or Transwelltrade mark systems) of human and murine cells. Purified RA synovial fibroblasts, but not healthy synovial or skin fibroblasts, induced hu/mu arthritis within 6 weeks. In-vitro secretion of murine and human interleukin(IL) 6, as well as murine tumour necrosis factor (TNF)-alpha, indicated cross activation between murine macrophages and human RA fibroblasts. Soluble-factor mechanisms proved more effective than cell-contact mechanisms. Purified RA fibroblasts can, alone, induce hu/mu SCID arthritis. The cytokine profile suggests that xenogeneic interaction between human fibroblasts and murine macrophages may determine the sequence of events leading to hu/mu arthritis. PMID- 11040072 TI - Immunogenicity of self antigens is unrelated to MHC-binding affinity: T-cell determinant structure of Golli-MBP in the BALB/c mouse. AB - The 'classical' myelin basic protein (MBP) exons belong to a much larger unit, termed the 'Golli-MBP' gene. Here we have examined the T-cell determinant structure of the Golli protein region in the BALB/c mouse. Golli p10-24, which was shown to have the strongest affinity for I-A(d), could not induce T-cell activation. Paradoxically, the poorer binding, overlapping p5-19 was effective at inducing T-cell proliferation. Thus, immunogenicity is not necessarily related to the MHC-binding affinity of self-peptides. In addition, MBP: p151-168-specific T cell clones responded only poorly to J37, a Golli-MBP protein, while MBP: 59-76 specific clones responded well to J37. PMID- 11040073 TI - Suppression of experimental autoimmune encephalomyelitis by intravenously administered polyclonal immunoglobulins. AB - Experimental autoimmune encephalomyelitis (EAE) was induced in Lewis rats either by active immunization with myelin basic protein (MBP) or by adoptive transfer using anti-MBP specific CD4(+)T cells. Treatment with human polyclonal immunoglobulins (IgG) effectively suppressed active EAE. Time-dependent experiments demonstrated that the effect of IgG was manifested only when treatment was given immediately after immunization; administration from day 7 after disease induction did not suppress the disease. In the adoptive transfer model of EAE, IgG had no effect in vivo. However, pretreatment in vitro of the antigen-specific T-cells with IgG inhibited their ability to mediate adoptive EAE, as it did in active EAE. Similarly, in vitro IgG pretreatment of the antigen specific T-cells suppressed the proliferative response to MBP. Fluorescent Activated Cell Sorter (FACS) analysis demonstrated the binding of IgG to activated T-cell lines that was inhibited by soluble Fc molecules. The differential effects of IgG on active EAE and on the adoptive transfer of EAE suggest that IgG in vivo can suppress disease by acting during the early phase of the immune response which involves naive T cells. The inhibition of T-cell proliferation and adoptive transfer of EAE by incubation of T cells in vitro appears to require higher concentrations of IgG than those obtained in vivo. PMID- 11040074 TI - Production of neurotrophins by activated T cells: implications for neuroprotective autoimmunity. AB - Neurotrophins (NTs) promote neuronal survival and maintenance during development and after injury. However, their role in the communication between the nervous system and the immune system is not yet clear. We observed recently that passively transferred activated T cells of various antigen specificities home to the injured central nervous system (CNS), yet only autoimmune T cells specific to a CNS antigen, myelin basic protein (MBP), protect neurons from secondary degeneration after crush injury of the rat optic nerve. Here we examined the involvement of NTs in T-cell-mediated neuroprotection, and the possible significance of the antigen specificity of the T cells in this activity. Analysis of cytokine and NT expression in various rat T cell lines showed that the T cells express mRNA for cytokines of Th1, Th2, and Th3 phenotypes. In addition, the T cells express mRNA and protein specific to nerve growth factor, brain-derived neurotrophic factor, NT-3, and NT-4/5. Antigen activation significantly increased NT secretion. Thus, reactivation of CNS autoimmune T cells by locally presented antigens to which they are specific can lead to enhanced secretion of NTs and possibly also of other factors in injured optic nerves. mRNA for TrkA, TrkB and p75 receptors was expressed in the injured nerve, suggesting that these specific receptors can mediate the effects of the T-cell-derived NTs. The neuroprotective effect of the passively transferred autoimmune anti-MBP T cells in injured optic nerves was significantly decreased after local applicaiton of a tyrosine kinase inhibitor known to be associated with NT-receptor activity. These results suggest that the neuroprotective effect of autoimmune T cells involves the secretion of factors such as NTs by the T cells reactivated by their specific antigen in the injured CNS. T cell intervention in the injured CNS might prove to be a useful means of promoting post-injury CNS maintenance and recovery, possibly via supply of NTs and other factors. PMID- 11040075 TI - Endogenous ecotropic murine leukemia viral (MuLV) envelope protein as a new autoantigen reactive with non-obese diabetic mice sera. AB - The identification and characterization of autoantigens associated with autoimmune IDDM (insulin dependent diabetes mellitus) would help to elucidate the pathogenic mechanism of this disease as well as to design antigen-based immunotherapy. Non-obese diabetic (NOD) mice have been used as the best model for studying the pathogenesis of human IDDM. To identify new autoantigens associated with IDDM, the lambda gt11-cDNA library from MIN6N8a, NOD-derived pancreatic beta cell line, was constructed and then candidate autoantigen clones were screened with prediabetic NOD sera. Nine positive clones were selected from 2x10(5)phage plaques. The nucleotide sequencing and homology searching showed that six of the nine positive clones had part of the endogenous ecotropic murine leukemia viral (MuLV) envelope gene. Nested deletion of this envelope gene revealed that the leucine zipper region in the transmembrane domain of MuLV envelope protein was the target epitope(s) reactive with prediabetic NOD mice sera. The prevalence of MuLV envelope protein-positive antibody in NOD mice was around 46%, while the non NOD mice strains including BALB/c, ICR, C57BL/6, and SJL/J mice did not produce this envelope protein-reactive antibody. The expression of endogenous ecotropic MuLV envelope gene in NOD mouse pancreas was distinct in those with severe insulitis. However, both prediabetic and diabetic NOD mice did not show the MHC class II-restrictive cellular autoimmunity against our purified recombinant envelope protein. In this study, we showed that the endogenous ecotropic MuLV envelope protein was a new autoantigen reactive with the activated NOD humoral immune system. PMID- 11040076 TI - Chronic myocarditis induced by T cells reactive to a single cardiac myosin peptide: persistent inflammation, cardiac dilatation, myocardial scarring and continuous myocyte apoptosis. AB - Recent recognition that an autoimmune myocarditis may precede, and result in, dilated cardiomyopathy has focused attention on immune mechanisms of myocardial injury. In this paper, we describe a model of chronic autoimmune myocarditis in the Lewis rat. The production of myocarditis has been previously described by this group and in brief is accomplished by a single tail vein infusion of activated T cells specific for a 17-amino acid peptide from rat cardiac myosin. In this report, animals were followed for approximately 6 months post-T-cell infusion. Hearts from animals which received cardiac myosin specific T cells all showed extensive fibrosis associated with ongoing inflammation. Apoptosis, identified by TdT-mediated dUTP nick end labelling (TUNEL), was identified as a mode of myocyte death in hearts with acute and chronic myocarditis but not in age and sex-matched controls. Immunohistochemistry was used to characterize the immune infiltrate and adhesion molecules in hearts with chronic myocarditis and these findings were compared to hearts with acute myocarditis. We propose that this rat model of chronic myocarditis mimics human disease, since inflammation results in ventricular dilatation and myocyte hypertrophy reminiscent of dilated cardiomyopathy. This model offers potential for further investigation of immune, functional and possible therapeutic aspects of autoimmune related cardiomyopathies. PMID- 11040077 TI - Identification and characterization of the antigen presenting cell in rat autoimmune myocarditis: evidence of bone marrow derivation and non-requirement for MHC class I compatibility with pathogenic T cells. AB - In the rat, autoimmune myocarditis can be produced by the infusion of activated myosin peptide specific, CD4(+), class II restricted, effector T cells. Whether antigen presenting cells (APCs), which interact with these effector T cells in the heart, are a fixed population of cells (resident dendritic, macrophage, or endothelial cells), or a dynamic bone marrow derived population has not yet been demonstrated in vivo. To study this question, bone marrow chimeras were generated using inbred Brown Norway (BN) rats, which are resistant to autoimmune myocarditis, and transplanting them after lethal irradiation with (LewisxBN) F1 bone marrow. BN rats differ at both MHC loci from the susceptible inbred Lewis rats. Two months after bone marrow transplantation, chimeric animals received Lewis T cells specific for a myocarditogenic peptide antigen. To characterize the cardiac APCs, immunohistochemistry using a battery of antibodies including Lewis specific and broadly reactive antibodies for both MHC class I and class II, was performed on chimeric hearts, with and without infused Lewis T cells, and non transplanted BN control hearts.All chimeric rats infused with allogeneic (Lewis), anti-cardiac myosin peptide effector T cells displayed the lesions of myocarditis. Myocarditis was not present in non-transplanted BN controls given either Lewis or F1 derived myocarditogenic T cells, nor in chimeric animals which did not receive myocarditogenic T cells, thus excluding graft vs host disease as the explanation for the inflammation in chimeric hearts with myocarditis. Marrow derived cells expressing both Lewis class I and class II MHC molecules were demonstrated on perivascular cells in the myocardium of all chimeric animals, and on infiltrating cells in chimeric animals with myocarditis. Cells expressing Lewis-specific MHC antigens were not detected in the non-transplanted BN controls. Furthermore, immunohistochemistry using broadly reactive antibodies demonstrated MHC class II on perivascular cells with a dendritic morphology in all hearts but not on endothelial cells or cardiac myocytes. These results support the hypothesis that in vivo, cardiac APCs which result in MHC class II restricted, T cell induced myocarditis are a dynamic bone marrow derived population and not a fixed population. In order to address the potential requirement of MHC class I for the initiation of autoimmune myocarditis, myocarditogenic T cells derived from either Lewis or DA(RP) rats were infused into a member of the other strain. These strains share common MHC class II genes but differ at the MHC class I loci. Myocarditis identical to that produced in the syngeneic animal was successfully transferred by the MHC class I mismatched T cells, but only after the recipient animal's native immune system was mildly suppressed. These results further support the primary role for professional antigen presentation via MHC class II restriction to the effector T cells at the initiation of autoimmune myocarditis in the heart.Together, these experiments confirm that activated effector T cells, in order to produce myocarditis, require MHC class II compatible APCs in the heart, that these APCs are bone marrow derived, and will endogenously take up and present local antigens in the target organ after bone marrow reconstitution. PMID- 11040078 TI - Linseed oil suppresses the anti-beta-2-glycoprotein-I in experimental antiphospholipid syndrome. AB - The immunomodulatory potential of a diet enriched with n-3 polyunsaturated fatty acids was analysed in naive mice with experimental antiphospholipid syndrome (APS) induced by active immunization with H-3, a human anti-beta-2-glycoprotein-I (anti-betaGPI) monoclonal antibody (mAb). Fetal loss and other clinical manifestations of APS were prevented in the group of APS mice upon exposure to the enriched n-3 diet compared to the control group. The titers of anti-betaGPI were significantly lower (in O.D. at 405 nm, 1.387+/-0.232 in comparison to non treated mice 0.637+/-0. 111, P< 0.05). The reduced titer of anti-betaGPI antibodies in the sera of the treated mice was associated with a reduced number of anti-betaGPI forming cells in cultured splenocytes (84+/-14, antibody-forming cells (AFC)/10(5)cells in comparison to 37+/-4 AFC/10(5)cells (P< 0.02).In addition to the suppression of the humoral response in mice with experimental APS fed with linseed oil, we also observed an inhibitory effect on the cellular response. The T-cell response to anti-betaGPI was lower in comparison to mice immunized with H-3 anti-betaGPI mAB, which were kept on a normal diet. These results indicate that polyunsaturated fatty acids may improve clinical and laboratory parameters of APS. The beneficial effects of diets enriched with n-3 should be further examined as a potential mode of therapy for patients with APS. PMID- 11040079 TI - Polymorphic glutathione S-transferase M1 is a risk factor of primary open-angle glaucoma among Estonians. AB - Primary open-angle glaucoma, the most common form of glaucoma is a slowly progressive atrophy of the optic nerve, characterized by loss of peripheral visual function and is usually associated with elevated intraocular pressure. The etiology and genetic risk factors of primary open-angle glaucoma are mostly unknown. The aim of this study was to find out whether the polymorphism at GSTM1, GSTM3, GSTT1 and GSTP1 loci is associated with increased susceptibility to glaucoma, because these polymorphic enzymes are susceptibility candidates for several diseases, including such eye disease as cataract. The phenotype of GSTM1 and GSTT1 was determined by ELISA and the genotype of GSTM3 and GSTP1 was detected by polymerase chain reaction. Four hundred and fifty two Estonians (250 glaucomas and 202 controls) participated in a case-control study. A significant association of the GSTM1 polymorphism with glaucoma was observed. The frequency of the GSTM1 positive individuals among the glaucoma group was significantly higher than in controls (60 vs. 45.0%) with odds ratio of 1.83 (95% CI 1.26 2.66;P = 0.002). The risk among the GSTM1 positive individuals of developing glaucoma was even higher in the case of smoking: 62.7% of smokers were GSTM1 positive in the glaucoma group while only 33.3% of smokers had GSTM1 positive phenotype in controls (OR = 3.36; 95% CI 1.49-7.56;P = 0.012). An association with a lower level of significance was also found with the GSTM3 gene. Four% of the 250 patients with POAG were identified as carriers of the GSTM3 BB genotype, a proportion which was slightly higher than the 1.0% for the controls (OR = 4.17; 95% CI 0. 90-19.24;P = 0.144). The frequencies of the GSTT1 and GSTP1 genotypes in both groups were not statistically different. The present study suggests that the GSTM1 polymorphism may be associated with increased risk of development of primary open-angle glaucoma. PMID- 11040080 TI - In vitro diffusion of mitomycin-C into human sclera after episcleral application: impact of diffusion time. AB - The purpose of this study was to investigate the impact of different diffusion times of mitomycin-C (MMC) on the intrascleral concentration vs depth profile of MMC in an experimental model. Scleral quadrants of eight human donor eyes were exposed to sponges soaked with MMC for an application time of 1 min. After irrigation with 40 ml saline, we allowed further diffusion of MMC in the sclera for 1, 5, 14 and 29 min until the specimens were further processed. A central 8 mm diameter scleral disk was horizontally dissected with a kryotome at -20 degrees C. MMC concentrations of six layers of 140 microm thickness were analysed by means of high-performance liquid chromatography. The MMC concentrations (microg g(-1)) of layer 1 were: 13.45+/- 5.9 (mean +/- S.D. at 2 min diffusion time), 7.6+/-2.5 (6 min diffusion), 5.6+/-3.1 (15 min diffusion) and 3.6+/-1.7 (30 min diffusion). The corresponding MMC concentrations of layer 6 were: 0.61+/ 0.48, 1.47 +/-0.66, 1.83+/-0.42 and 2.98+/-0.97 microg g(-1). The superficial concentration of intrascleral MMC decreased with increasing diffusion time, the deep concentrations increased. After 30 min of diffusion time, equal concentrations of MMC were found in all layers. Even with current low-dose application regimens of MMC the concentrations in the inner side of the sclera rapidly increase beyond the limits of the therapeutic range. Owing to this fast diffusion of MMC, the only means of reducing ciliary body concentrations of MMC is to reduce the dose. PMID- 11040081 TI - Expression of Sonic hedgehog and retinal opsin genes in experimentally-induced myopic chick eyes. AB - The purpose of this study was to evaluate changes in the expression of different genes in chick retinal tissues after induction of experimental myopia and to evaluate the roles of these genes in the regulation of postnatal eye growth and myopia. Form-deprivation using occlusive goggles and hyperopic defocus by negative spectacle lenses were used to induce myopia in hatched chicks. Expression levels of Sonic hedgehog, its receptor complex, and other retinal cell genes were evaluated by semi-quantitative reverse transcription-polymerase chain reaction. Levels of Sonic hedgehog protein were further evaluated by Western blot analysis. The induction of myopia caused significant increase in expression of Sonic hedgehog mRNA and protein and increased expression of blue and red opsin mRNA. In contrast, the expression of mRNA for Sonic hedgehog receptor complex (Patched-Smoothened), rhodopsin, vimentin, green opsin, violet opsin, and HPC-1 were unaffected by the induction of myopia. The increase in expression of Sonic hedgehog in chick retinas in experimentally-induced myopia suggests involvement in the retina control of postnatal eye growth. Furthermore, Sonic hedgehog may influence the expression of blue and red opsins under myopic conditions. PMID- 11040082 TI - Retinal function is improved in a murine model of a lysosomal storage disease following bone marrow transplantation. AB - Mucopolysaccharidoses are heritable lysosomal storage diseases caused by deficiencies in acid hydrolases involved in the sequential degradation of complex glycosaminoglycans (GAGs). In many mucopolysaccharidoses, GAGs accumulate in the retinal pigment epithelial (RPE) cells of the eye resulting in pronounced lysosomal distension. It is not clear how the progressive accumulation of GAGs affects retinal function. Bone marrow transplantation (BMT) is a relatively effective therapy for many lysosomal storage diseases and can result in a dramatic reduction in lysosomal distention in the RPE. Although effective at reducing lysosomal storage, it is not clear how effective syngeneic BMT is at treating retinal dysfunction. Here we show that there is a progressive decrease in the amplitudes of both the dark-adapted (rod-cone) and light-adapted (cone dominated) flash electroretinograms (ERG) between 8 and 20 weeks of age in a murine model of mucopolysaccharidosis type VII (MPS VII). By 20 weeks, the average dark-adapted b-wave amplitude was 118 microV in MPS VII mice as compared to 469 microV in normal mice of the same strain. MPS VII mice receiving syngeneic BMT at 4 weeks of age have reduced lysosomal storage in retinal pigment epithelial cells and normal ERG amplitudes at 20 weeks of age. Retinal function is impaired in untreated 8 week old MPS VII mice. Following BMT at 8 weeks, rod cone- and cone-dominated responses recovered to within the normal range by 20 weeks of age. These data demonstrate the temporal pattern of retinal dysfunction in the MPS VII mouse and indicate that BMT can reduce the lysosomal storage and improve retinal function. PMID- 11040083 TI - Treatment of corneal neovascularization with dietary isoflavonoids and flavonoids. AB - The purpose of this study was to investigate the use of dietary isoflavonoids and flavonoids for the treatment of ocular neovascularization. Corneal blood vessels were induced by intrastromal implantation of pellets containing bFGF. Isoflavonoids and flavonoids (Genistein, Fisetin and Luteolin) were dissolved in a microemulsion to increase bioavailability and applied topically in concentrations between 0.5 and 1 ng ml(-1). Corneal neovascularization was quantified under the microscope. In comparison to control eyes, all three substances significantly inhibited corneal neovascularization (P < or = 0.05). Fisetin had the strongest effect followed by Genistein and Luteolin. No significant topical side effects were observed. We concluded that the isoflavonoid Genistein and two structurally related flavonoids are potent inhibitors of corneal angiogenesis in vivo. The wide distribution of the flavonoids in the plant kingdom together with the presented results suggests that flavonoids may contribute to the preventive effect of a plant-based diet on neovascular disease of the eye. PMID- 11040084 TI - Lens epithelial cell proliferation in human posterior capsule opacification specimens. AB - In previous in vitro studies on capsular bags it was shown that, after a sham extracapsular cataract extraction (ECCE) on human donor eyes, lens epithelial cells (LECs) show, in the short term, a dramatically elevated mitotic activity as compared to that in the intact lens. The long term in vivo proliferation of LECs in human lenses after ECCE and intraocular lens (IOL) implantation has not been studied until now. In the present study, the mitotic activity of LECs in human post-mortem eyes with posterior capsule opacification (PCO) was investigated. Human lenses with signs of PCO were dissected from donor eyes and incubated in MEM, supplemented with fetal calf serum, for 1 day (n = 10) or 7 days (n = 9). Six additional specimens were cultured for 7 days after removal of the IOL and lens fibres. After the incubation period, mitotic activity was estimated using the BrdU procedure and the Ki67 proliferating cell marker. The mean number of BrdU-positive nuclei in the intact PCO specimens was at a level of 7.5 (day 1) and 6.5 (day 7). Removal of the IOL and the lens fibres leads to a ten-fold increase in BrdU positive cells (mean = 84.5). No correlation with donor age was found. The Ki67 observations corroborate the BrdU results. The results demonstrate that after an initial rise in proliferative activity, as shown in the capsular bag model, the mitotic activity of LECs returns to a rate comparable to that in intact cultured non-cataractous lenses. As in control lenses, removal of lens fibres significantly elevated the proliferative activity of the remaining LECs. Suppression by newly formed differentiated lens fibres in the in vivo capsular bag may be responsible for this return to control levels of mitotic activity of LECs in the PCO specimens. PMID- 11040085 TI - A herpes simplex virus type 1 vector as marker for retrograde neuronal tracing: characterization of lacZ transcription and localization of labelled neuronal cells in sensory and autonomic ganglia after inoculation of the anterior segment of the eye. AB - Herpes simplex virus type 1 (HSV-1) is a human, neurotropic pathogen which also can infect experimental animals. Much interest has been focused on genetic modification of HSV-1 so that it can be used as a vector for gene delivery and for tracing neuronal connections. For expression of a foreign gene inserted into the HSV-1 genome, both the site of insertion and the promoter activity are important. We have used a previously described HSV-1 vector, KOS/58, to demonstrate that the beta-galactosidase gene inserted together with a neurofilament L promoter into the coding region of the glycoprotein C (gC) gene is under the control of the foreign promoter rather than under that of the gC gene. This was performed by isolation of RNA from infected, neuron-like PC12 cells and Northern blotting using probes from various regions of the modified part of the genome. The vector was then inoculated in the cornea, subconjunctivally, or into the anterior chamber of the mouse eye. Whole mounts of the trigeminal, superior cervical and pterygopalatine ganglions were stained for beta-galactosidase. The localization of labelled neurons was consistent with retrograde axonal transport as the principal way of neuronal infection indicating that KOS/58 could be used as a retrograde tracer. The position of the labelled cells suggests a somatotopic organization of the mouse trigeminal and superior cervical ganglion similar to that of rats and rabbits. PMID- 11040086 TI - The effect of hydration and matrix composition on solute diffusion in rabbit sclera. AB - There is increasing interest in the possibility for drug delivery into the vitreous humor across the conjunctiva and sclera as an alternative route to the conjunctiva-cornea pathway. As a preliminary to human studies we have investigated the influence of scleral composition and hydration on solute transport in the rabbit sclera. Intermuscular sclera was excised from adult New Zealand rabbits. Tissue samples were either examined directly (controls), digested using chondroitinase ABC or crosslinked using glutaraldehyde. The effect of these treatments on the ultrastructural appearance of the sclera was assessed. Diffusion and partition coefficients for solutes of different molecular weights [sodium chloride (23 MW),(14)C sucrose (342 MW) and dextran-fluoresceins (3, 10, 40 and 70 kDa)] were measured in relation to tissue treatment. The results were used to determine the effect of tissue structure and composition on solute movement. We have found that: (1) diffusion and partition coefficients are sensitive to solute MW, decreasing as MW increases; (2) diffusion and partition coefficients are sensitive to tissue hydration, increasing as hydration increases; (3) crosslinking of the sclera by glutaraldehyde reduced the partition coefficients significantly for solutes with MW over 3 kDa; and (4) removal of glycosaminoglycans has only a small effect on either diffusion or the partition coefficient. PMID- 11040087 TI - Aldose reductase does catalyse the reduction of glyceraldehyde through a stoichiometric oxidation of NADPH. AB - In order to define the ability of bovine lens aldose reductase (ALR2) to generate polyols from aldoses, the quantitative determination of glycerol in the presence of glyceraldehyde was performed by gas chromatography after derivatization with trifluoroacetic anhydride. The proposed method appears to be useful in quantifying low amounts of glycerol in the presence of relatively high concentrations of glyceraldehyde and in following glycerol formation in enzyme assay conditions. The generation of one equivalent of glycerol in the presence of ALR2, is paralleled by the oxidation of one equivalent of NADPH. A similar result was obtained when S-glutathionyl-modified ALR2 was used, instead of the native enzyme, as a catalyst of glyceraldehyde reduction. Sorbinil, a classical ALR2 inhibitor, present in the enzyme assay mixture, inhibits to the same extent both NADPH oxidation and glycerol formation. The demonstration of the stoichiometric ratio of 1:1 occurring in the presence of bovine lens ALR2 between the synthesis of glycerol from D, L -glyceraldehyde and the oxidation of NADPH, rules out doubts concerning the ability of the enzyme to catalyse the reduction of aldoses to the corresponding polyalcohols. Possible autooxidation processes of glyceraldehyde, in the enzyme assay conditions, appear to be irrelevant with respect to the enzyme-catalysed reduction of the aldose. This would indicate that the spectrophotometric monitoring of NADPH oxidation at 340 nm, in the presence of ALR2, is a reliable method to assay the enzyme activity. PMID- 11040088 TI - Quantitative image analysis of laser-induced choroidal neovascularization in rat. AB - Rodent models of laser-induced choroidal neovascularization (CNV) are now extensively used to identify angiogenic proteins, determine the role of specific genes with knockout mice, and evaluate the efficacy and safety of anti-angiogenic therapies. CNV is typically evaluated by fluorescein angiography or vascular endothelial cell labeling in histologic sections. The current study examined an alternative method using high molecular weight FITC-dextran (MW 2 x 10(6)) for high resolution angiography in RPE-choroid-sclera flat mounts. At 24 hr after lasering, the lesions appeared as a circular weakly fluorescent area of approximately equal diameter to the laser spot. No FITC-dextran labeled blood vessels were visible in the lesion at day 1. Three days after lasering, 47% of the lesions showed FITC-dextran labeling indicative of CNV. The incidence (71%) and extent of CNV increased by day 6, and by day 10 all lesions were vascularized, and the maximal area was attained. No significant change followed day 10, and the neovascular area remained constant through day 31. The highest rate of blood vessel growth (between 3 and 10 days after laser) correlates with the peak expression of VEGF, bFGF, and their receptors shown in previous studies. Morphologic analysis of flat mounts and histologic sections showed that the neovascular plexus in most lesions originates from deeper choroidal vessels in the center of the lesion, grows towards the neural retina, then branches circumferentially to anastamose with uninjured choriocapillaris. The microvessels in these lesions are broad and flat, similar to normal choriocapillaris. In a separate study, rats were treated daily with the angiostatic corticosteroid dexamethasone (20-500 microg kg(-1)day(-1)), and CNV was examined at day 10 in FITC-dextran labeled flat mounts and histologic sections. Dexamethasone dose dependently inhibited CNV, and its highest dose inhibited approximately 95% of CNV labeled by FITC-dextran and resulted in lesions with no detectable Factor VIII immunostaining. High resolution angiography with FITC-dextran is reproducible and quantifiable, and it may accelerate the discovery of therapeutic agents that modulate choroidal neovascularization. PMID- 11040089 TI - Endothelin-1 induces nitric oxide synthase-2 expression in human non-pigmented ciliary epithelial cells. PMID- 11040090 TI - Detection of PCR products using PNA strand invasion. AB - The unique ability of homopyrimidine peptide nucleic acid (PNA) to strand invade homopurine sites of duplex DNA offers a potential alternative to existing techniques for rapid detection of PCR products. From gel shift studies, PNA was found to specifically strand invade homopurine sites that had been incorporated into an amplicon during the PCR cycle. This was achieved by adding a homopyrimidine sequence to the 5'-terminus of a PCR primer. The position of the strand invasion sites at the termini of the DNA duplex offers kinetic advantages for PNA strand invasion, since the termini of DNA duplexes are known to be unwound. This unwound state was demonstrated using a novel assay that determined single-stranded regions within the amplicon. The presence of the PNA moiety in strand invasion complexes was confirmed by a novel electroblot, an Enzyme Linked Nucleic Acid assay and by an increase in stability as demonstrated by T(m)studies with the Idaho RapidCycler. Since the strand invasion sites can be controlled through selection of the homopurine sequence there is great flexibility for designing strand invasion motifs unique to a particular PCR amplicon, thus providing a huge potential for differentiating and detecting multiplex PCR products. PMID- 11040091 TI - 5 S ribosomal spacer sequences of some filarial parasites: comparative analysis and diagnostic applications. AB - We have amplified by PCR the sequences of the 5 S ribosomal spacer of Setaria labiatopapillosa and Foleyella furcata. After sequencing, these sequences have been compared with those of Dirofilaria immitis and Dirofilaria repens. Two major goals have been achieved: (i) the establishment of a multiplex PCR-based diagnostic assay, applicable to identify the four species in vertebrate and invertebrate hosts; (ii) the identification, in S. labiatopapillosa and F. furcata, of a canonical spliced leader 1 (SL1) sequence, so confirming that only D. repens, of the filarial parasites so far studied, shows a peculiar SL1 sequence. The PCR assay here developed and the analysis of the 5 S ribosomal spacer, can further improve both epidemiological and molecular analysis of these filarial species. PMID- 11040092 TI - DNA fingerprints of Trichinella as revealed by restriction fragment length polymorphism and single-strand conformational polymorphism (RFLP-SSCP). AB - A method was developed for gene fingerprinting, combining the principles of restriction fragment length polymorphism and single-strand conformational polymorphism (PCR-RFLP-SSCP). Taking advantage of this method, we analysed the genotypes of 20 isolates from five species of Trichinella (Trichinella spiralis, Trichinella britovi, Trichinella nativa, Trichinella murrelli and Trichinella pseudospiralis) and two uncertain genotypes (Trichinella T6 and Trichinella T8). Target genes for the analysis included three kinds of random amplified polymorphic DNA (RAPD), the gene encoding 43 kDa excretory-secretory protein, the gene encoding mitochondrial cytochrome c -oxidase subunit I and the gene of 18 S rRNA. The genotype revealed by this method was in good concordance with the taxonomy of Trichinella (six species and two uncertain genotypes which is currently accepted based on morphology, isozyme pattern and reproductive isolation). More important, this method revealed intraspecies polymorphism among geographical isolates of T. spiralis and T. britovi. PMID- 11040093 TI - Mutation analysis in a small cohort of New Zealand patients originating from the United Kingdom demonstrates genetic heterogeneity in familial hypercholesterolemia. AB - Familial hypercholesterolemia (FH) and familial defective apolipoprotein B-100 (FDB) are relatively common lipid disorders caused by mutations in the low density lipoprotein receptor (LDLR) and apolipoprotein B (apo B) genes, respectively. Molecular analysis at these loci was performed in eight New Zealand subjects with clinical features of heterozygous FH. Utilization of an in vitro lymphocyte receptor assay demonstrated normal receptor function in four patients, three of whom screened positive for the founder-type apo B mutation, R3500Q, causing FDB. Four patients with reduced LDLR function, consistent with heterozygous FH, revealed three previously documented mutations in exons 3 (W66X), 6 (C292Y) and 7 (G322S) of the LDLR gene and, a novel 2-bp deletion (TC or CT) after nucleotide 1204 (or 1205) in exon 9. The remaining patient was found to be FH/FDB negative after extensive mutation screening using both denaturing gradient gel electrophoresis and heteroduplex-single strand conformation polymorphism analysis. Haplotype analysis at the LDLR and apo B loci finally excluded the likelihood that mutations in these two genes underlie the FH phenotype in the molecularly uncharacterized New Zealand family originating from the United Kingdom. This family represents a valuable source of material for future genetic dissection of autosomal dominant hypercholesterolemia (ADH), shown to be a heterogeneous disease through molecular analysis. PMID- 11040094 TI - Sheep poxvirus identification from clinical specimens by PCR, cell culture, immunofluorescence and agar gel immunoprecipitation assay. AB - Some 40 clinical specimens of skin lesions from sheep pox suspected cases were investigated by four different diagnostic assays: PCR, virus isolation in lamb testis cell cultures, direct immunofluorescent assay (DIFA) and antigen detecting agar gel immune precipitation test (AGIPT). All the specimens were positive by PCR and virus isolation, 29 were positive by DIFA and 16 by AGIPT. Using virus isolation on cell cultures as the gold standard, the PCR sensitivity was 100%, while that of DIFA and AGIPT was 73% and 40%, respectively. Skin samples with orf lesions or normal skin biopsies were PCR-negative. Cross-reactions with orf virus were observed in three samples only in the AGIPT assay. The PCR described combines high specificity and sensitivity with speed. PCR was therefore shown to be the method of choice for sheep poxvirus diagnosis directly from clinical specimens. PMID- 11040095 TI - Comparative molecular analysis of erythromycin-resistance determinants in staphylococcal isolates of poultry and human origin. AB - The ermA, ermB, ermC and msrA/msrB genes were detected in multidrug-resistant Staphylococcus spp. strains by PCR. Among 25 human clinical staphylococcal isolates the ermA, ermB, ermC and the msrA/msrB genes were detected in 88, 72, 4 and 100% of the strains, respectively. Among 24 poultry isolates the ermA, ermB, ermC and the msrA/msrB genes were detected in 100, 16.6, 50 and 12.5% of the strains, respectively. The ermA gene was found exclusively on the chromosome, whereas the ermC gene was found on 2.4-4.2 kb plasmids. Restriction fragment length polymorphism (RFLP) analysis of the ermA gene with Eco RI revealed five patterns (25.0, 21.0, 10.5, 6.2 and 4. 8 kb) for the clinical strains and two (8.0 and 6.2 kb) for the poultry strains. The 6.2 kb RFLP pattern, in both the poultry and human clinical isolates, indicates a common lineage for the ermA gene. PMID- 11040096 TI - Rapid and specific detection of PCR products using light-up probes. AB - Newly developed light-up probes offer an attractive tool for PCR product detection. The light-up probe, which consists of a thiazole orange derivative linked to a peptide nucleic acid oligomer, hybridizes specifically to complementary nucleic acids. Upon hybridization the thiazole orange moiety interacts with the nucleic acid bases and the probe becomes brightly fluorescent. This eliminates the need to separate bound from unbound probes and reduces the risk of cross contamination during sample handling. We demonstrate here the applicability of light-up probes in two different PCR assays, one directed towards the human beta-actin gene and the other towards the invA gene of Salmonella. The probes do not interfere with the PCR reaction and can either be included in the sample mixture or added after completed amplification. The specificity of the probe is found to be excellent: a single-base mismatch in the target sequence is sufficient to prevent probe binding as indicated by the lack of fluorescence increase. Furthermore, a clear correlation is found between the intensity of gel bands and the measured probe fluorescence in solution, which suggests that the amount of PCR products can be quantified using light-up probes. PMID- 11040097 TI - Mitochondrial K(ATP) channels: probing molecular identity and pharmacology. PMID- 11040098 TI - Bioinformatics analysis of gene banks provides a treasure trove for the functional genomist. PMID- 11040099 TI - Building a better mouse model. PMID- 11040100 TI - Prevention of abnormal sarcoplasmic reticulum calcium transport and protein expression in post-infarction heart failure using 3, 5-diiodothyropropionic acid (DITPA). AB - Heart failure of diverse causes is associated with abnormalities of sarcoplasmic reticulum (SR) Ca(2+)transport. The purpose of this study was to determine whether the thyroid hormone analogue, 3,5-diiodothyropropionic acid (DITPA), prevents abnormal Ca(2+)transport and expression of SR proteins associated with post-infarction heart failure. New Zealand White rabbits were randomly assigned to circumflex artery ligation or sham operation, and to DITPA administration (3.75 mg/kg/day) or no treatment in a two-by-two factorial design. After 3 weeks, echo-Doppler and LV hemodynamic measurements were performed. From ventricular tissue, single myocyte shortening and relaxation were determined, and Ca(2+)transport was measured in homogenates and SR-enriched microsomes. Levels of mRNA and protein content were determined for the SR Ca(2+)-ATPase (SERCA2a), phospholamban (PLB), cardiac ryanodine receptor (RyR-2) and calsequestrin. The administration of DITPA improved LV contraction and relaxation and improved myocyte shortening in infarcted animals. The improvements in LV and myocyte function were associated with increases in V(max)for SR Ca(2+)transport in both homogenates and microsomes. Also, DITPA prevented the decrease in LV protein density for SERCA2a, PLB and RyR-2 post-infarction, without measurable changes in mRNA levels. The thyroid hormone analogue, DITPA, improves LV, myocyte and SR function in infarcted hearts and prevents the downregulation of SR proteins associated with post-infarction heart failure. The specific effects of DITPA on post-infarction SR Ca(2+)transport and the expression of SR proteins make this compound a potentially useful therapeutic agent for LV systolic and/or diastolic dysfunction. PMID- 11040102 TI - Thyroid hormone regulation of myocardial Na/K-ATPase gene expression. AB - Employing published methods for isolation of cardiac myocyte nuclei from adult rat ventricular myocardium with the use of mechanical disruption without digestive enzymes, we obtained transcriptionally active cardiac myocyte nuclei with sufficient yield and purity. The relative content of Na/K-ATPase subunit mRNAs (alpha 1, alpha 2, and beta 1) in ventricular myocardium of euthyroid rats closely matched the relative rates of transcription of the respective subunit genes determined by nuclear run-on assay. Treatment of hypothyroid rats with T(3)to elicit hyperthyroidism was associated with 2.9-, 7.5-, and seven-fold increases in the contents of alpha 1-, alpha 2, beta 1-mRNAs, respectively. In contrast, rates of transcription of the subunit genes were not changed significantly by T(3), while transcription of the 18 S ribosomal gene was stimulated identical with three-fold by the treatment. A quantitative reverse transcription-polymerase chain reaction assay for measurement of primary RNA transcripts of the beta 1 gene was developed employing a rat genomic DNA fragment that contains the first exon and part of the first intron of the beta 1 gene. The relative abundance of beta 1 primary transcripts did not change in RNA isolated from hypothyroid, euthyroid, and hyperthyroid rats. It is concluded that: (1) The relative contents of Na/K-ATPase subunit mRNAs in euthyroid adult myocardium is primarily controlled at the transcriptional level, and (2) T(3)-induced increases in the contents of Na/K-ATPase subunit mRNAs in the heart is not associated with increased rates of transcription of the subunit genes, and the effect is mediated at the post-transcriptional level. PMID- 11040101 TI - Induction of VEGF gene transcription by IL-1 beta is mediated through stress activated MAP kinases and Sp1 sites in cardiac myocytes. AB - Interleukin-1 beta (IL-1 beta) is a multipotent cytokine participating in a variety of cardiovascular diseases. In this study, we examined the effects of IL 1 beta on the expression of vascular endothelial cell growth factor (VEGF) and pursued the molecular mechanisms underlying this effect. Treatment of cultured neonatal rat cardiac myocytes with IL-1 beta increased the levels of VEGF mRNA in a time- and a concentration-dependent manner. These effects were completely abolished by SB203580 and SB202190 (p38 MAPK inhibitors) but not by PD98059 (MEK1 inhibitor), calphostin C (protein kinase C inhibitor), or genistein (tyrosine kinase inhibitor). While IL-1 beta phosphorylated c-Jun N-terminus protein kinase (JNK) rapidly and transiently, the effect of IL-1 beta on p38 mitogen-activated protein kinase (MAPK) was gradual and persistent. Transient transfection assays showed that IL-1 beta increases the transcription from the VEGF promoter. A series of 5;-deletion and site-specific mutation analyses indicated that IL-1 beta as well as overexpression of p38 MAPK and JNK activate VEGF promoter activity through two G+C-rich sequences located at -73 and -62. Electrophoretic mobility shift and supershift assays showed Sp1 and Sp3 proteins specifically bind to the G+C-rich sequences. The half-life of VEGF mRNA was significantly increased in cells treated with IL-1 beta. Together, these results indicate that IL-1 beta induces VEGF gene expression at both transcriptional and post transcriptional levels, and IL-1 beta evokes p38 MAPK and JNK signalings, which in turn stimulate the transcription of the VEGF gene through Sp1-binding sites. These findings suggest the role of IL-1 beta as a cytokine inducing VEGF in cardiac myocytes, and imply that activation of stress-activated MAP kinases regulate Sp1 sites-dependent transcription. PMID- 11040103 TI - Immunogold-labeled L-type calcium channels are clustered in the surface plasma membrane overlying junctional sarcoplasmic reticulum in guinea-pig myocytes implications for excitation-contraction coupling in cardiac muscle. AB - Ca(2+) release through ryanodine receptors, located in the membrane of the junctional sarcoplasmic reticulum (SR), initiates contraction of cardiac muscle. Ca(2+)influx through plasma membrane L-type Ca(2+)channels is thought to be an important trigger for opening ryanodine receptors ("Ca(2+)-induced Ca(2+) release"). Optimal transmission of the transmembrane Ca(2+)influx signal to SR release is predicted to involve spatial juxtaposition of L-type Ca(2+)channels to the ryanodine receptors of the junctional SR. Although such spatial coupling has often been implicitly assumed, and data from immunofluorescence microscopy are consistent with its existence, the definitive demonstration of such a structural organization in mammalian tissue is lacking at the electron-microscopic level. To determine the spatial distribution of plasma membrane L-type Ca(2+)channels and their location in relation to underlying junctional SR, we applied two high resolution immunogold-labeling techniques, label-fracture and cryothin sectioning, combined with quantitative analysis, to guinea-pig ventricular myocytes. Label-fracture enabled visualization of colloidal gold-labeled L-type Ca(2+)channels in planar freeze-fracture electron-microscopic views of the plasma membrane. Mathematical analysis of the gold label distribution (by nearest neighbor distance distribution and the radial distribution function) demonstrated genuine clustering of the labeled channels. Gold-labeled cryosections showed that labeled L-type Ca(2+)channels quantitatively predominated in domains of the plasma membrane overlying junctional SR. These findings provide an ultrastructural basis for functional coupling between L-type Ca(2+)channels and junctional SR and for excitation-contraction coupling in guinea-pig cardiac muscle. PMID- 11040104 TI - Calcium handling and role of endothelin-1 in monocrotaline right ventricular hypertrophy of the rat. AB - We investigated the role of endothelin-1 (ET-1) in right ventricular function and intracellular Ca(2+)(Ca(2+)(i)) handling of isolated perfused rat hearts with right ventricular hypertrophy induced by monocrotaline (50 mg/kg). Nine weeks after monocrotaline (n=9) or saline (control n=9) treatment, hearts were perfused isovolumically at 37 degrees C and right ventricular function (fluid-filled balloon), right ventricular intracellular Ca(2+) transients (aequorin bioluminescence method) and the effects of ET-1 were determined. Monocrotaline treated rats developed considerable right ventricular hypertrophy (right ventricular weight:body weight ratio: 1.07+/-0.13 v. 0.60+/-0.03 in controls P<0.05) and these hearts generated higher right ventricular systolic and diastolic pressure, but similar systolic and diastolic wall stress, indicating a compensated functional state. Hypertrophied hearts demonstrated a prolonged duration of isovolumic contraction (time to 90% decline from peak: 105+/-1 v 89+/ 4 ms at 3 m M extracellular Ca(2+) P<0.05), but neither the time to peak pressure (71+/-3 ms) nor time to peak light (25+/-3 ms) were different from controls. The increased duration of contraction correlated with a similar prolongation of the Ca(2+)transient (time to 90% decline from peak: 72+/-4 v 50+/-3 ms P<0.05), indicating a reduced rate of Ca(2+)sequestration in hypertrophic right ventricles. Peak systolic intracellular Ca(2+)was similar in control and hypertrophied hearts (1.04+/-0.02 and 0.99+/-0.02 microM, P>0.05, n=6). ET-1 (1 300 p M) affected neither the time course of right ventricular contraction nor that of the Ca(2+)transient or peak systolic Ca(2+)concentrations. These data are the first measurements of right ventricular Ca(2+)transients in beating normal and hypertrophic hearts. We conclude that ET-1 plays no role in compensated hypertrophy because it affected neither right ventricular function nor intracellular Ca(2+)handling in this model. PMID- 11040105 TI - Modulation of potassium channels in the hearts of transgenic and mutant mice with altered polyamine biosynthesis. AB - Inward rectification of cardiac I(K1)channels was modulated by genetic manipulation of the naturally occurring polyamines. Ornithine decarboxylase (ODC) was overexpressed in mouse heart under control of the cardiac alpha -myosin heavy chain promoter (alpha MHC). In ODC transgenic hearts, putrescine and cadaverine levels were highly elevated ( identical with 35-fold for putrescine), spermidine was increased 3.6-fold, but spermine was essentially unchanged. I(K1)density was reduced by identical with 38%, although the voltage-dependence of rectification was essentially unchanged. Interestingly, the fast component of transient outward (I(to,f)) current was increased, but the total outward current amplitude was unchanged. I(K1)and I(to)currents were also studied in myocytes from mutant Gyro (Gy) mice in which the spermine synthase gene is disrupted, leading to a complete loss of spermine. I(K1)current densities were not altered in Gy myocytes, but the steepness of rectification was reduced indicating a role for spermine in controlling rectification. Intracellular dialysis of myocytes with putrescine, spermidine and spermine caused reduction, no change and increase of the steepness of rectification, respectively. Taken together with kinetic analysis of I(K1)activation these results are consistent with spermine being a major rectifying factor at potentials positive to E(K), spermidine dominating at potentials around and negative to E(K), and putrescine playing no significant role in rectification in the mouse heart. PMID- 11040106 TI - Altered expression of proteins of metabolic regulation during remodeling of the left ventricle after myocardial infarction. AB - Non-infarcted myocardium after coronary occlusion undergoes progressive morphological and functional changes. The purpose of this study was to determine whether non-infarcted myocardium exhibits (1) alteration of the substrate pattern of myocardial metabolism and (2) concomitant changes in the expression of regulatory proteins of glucose and fatty acid metabolism. Myocardial infarction was induced in rats by ligation of the left coronary artery. One day and eight weeks after coronary occlusion, glucose and palmitate oxidation were measured. Expression of selected proteins of metabolism were determined one day to 12 weeks after infarction. One day after coronary occlusion no difference of glucose and palmitate oxidation was detectable, whereas after eight weeks, glucose oxidation was increased (+84%, P<0.05) and palmitate oxidation did not change significantly (-19%, P=0.07) in infarct-containing hearts, compared with hearts from sham operated rats. One day after coronary occlusion, myocardial mRNA expression of the glucose transporter GLUT-1 was increased (+86%, P<0.05) and the expression of GLUT-4 was decreased (-28%, P<0.05) in surviving myocardium of infarct-containing hearts. Protein level of GLUT-1 was increased (+81%, P<0.05) and that of GLUT-4 slightly, but not significantly, decreased (-16%, P=NS). mRNA expressions of heart fatty acid binding protein (H-FABP), and of medium chain acyl-CoA dehydrogenase (MCAD), were decreased by 36% (P<0.05) and 35% (P=0. 07), respectively. Eight weeks after acute infarction, the left ventricle was hypertrophied and, at this time-point, there was no difference in the expression of GLUT-1 and GLUT-4 between infarcted and sham-operated hearts. However, myocardial mRNA and protein content of MCAD were decreased by 30% (P<0.01) and 27% (P<0.05), respectively. In summary, in surviving myocardium, glucose oxidation was increased eight weeks after coronary occlusion. Concomitantly, mRNA and protein expression of MCAD were decreased, compatible with a role of altered expression of regulatory proteins of metabolism in post-infarction modification of myocardial metabolism. PMID- 11040107 TI - Alterations in cardiac function and gene expression during autoimmune myocarditis in mice. AB - Although myocarditis has been implicated in the pathogenesis of heart failure, a definitive relationship between myocardial inflammation, cardiac dysfunction, and changes in myocyte gene expression has not been established. In this study, we examined the hypothesis that myocardial inflammation and replacement fibrosis following an autoimmune response can progress to cardiac dysfunction and may result in progression to the heart failure phenotype. SWXJ mice were immunized with cardiac myosin on day 0 and day 7, in order to induce an autoimmune response to the myosin protein. Cardiac catheterization via the right carotid artery was performed on days 14, 21, 28, 35, and 42, using a 1.4F Millar transducer-tipped catheter. Hearts were weighed, and cross-sections were cut and stained with either haematoxylin and eosin or Masson's trichrome, in order to identify areas of inflammation and/or fibrosis. Myocardial gene expression was determined by Northern blot analysis. In mice with histological evidence of myocarditis, the heart weight/body weight ratio increased beginning on day 14, and cardiac function decreased beginning on day 21. Myocardial inflammation was accompanied by significant fibrosis beginning on day 21. Quantitation of mRNA showed expression of ventricular atrial naturietic factor, as well as a decrease in myosin heavy chain alpha, beginning on day 21. These data demonstrate that autoimmune inflammation of the heart results in significant cardiac dysfunction, leading to phenotypic alterations similar to those demonstrated in human heart failure and animal models of heart failure. PMID- 11040108 TI - Characterization of the sarcoplasmic reticulum k(+) and Ca(2+)-release channel ryanodine receptor-in human atrial cells. AB - Since the role of sarcoplasmic reticulum (SR) in the E-C coupling of mammalian atrial cells has long been a subject of debate, biochemical, electrophysiological and immunological assays were performed in order to define and compare the properties of the Ca(2+)-release channel-ryanodine receptor (RyR)-from atrial and ventricular tissues. Cardiac SR preparations from human, canine and ovine tissues were compared using [(3)H]ryanodine binding, channel reconstitution into planar lipid bilayers and Western blot analysis involving RyR antibodies. [(3)H]ryanodine binding assays revealed a K(d)value of; 2.5 n M for all investigated cardiac tissues. Bound [(3)H]ryanodine was Ca(2+)-dependent with similar EC(50)values of 0.43, 0.49 and 0.79 microM for human atrium, canine ventricle and ovine atrium, respectively. However the density of binding sites was 4.5 times lower in atrial than in ventricular tissues. Beyond the presence of selective K(+)channels (gamma=188 pS) recorded in the SR enriched fraction of human atrium, the activity of a large conducting (gamma=671 pS) cationic channel was also observed. The latter displayed typical characteristics of Ca(2+)-release channels which were activated by 10 microM free [Ca(2+)] and 2 m M ATP. Western blot analysis revealed the presence of the RyR2 isoform in atrial and ventricular samples whereas no immunoreactivity was detected with specific RyR1 and RyR3 antibodies. Our results, obtained at the molecular level, are consistent with the presence of functional SR in human atrial cells. The human atrial Ca(2+)-release channel displays binding and regulating properties typical of the RyR2 isoform. PMID- 11040109 TI - Sulphonylurea-sensitive channels and NO-cGMP pathway modulate the heart rate response to vagal nerve stimulation in vitro. AB - Sulphonylurea-sensitive K(+)channels (K(ATP)) have been implicated in the release of acetylcholine (ACh) from the vagus nerve in the heart. Our aim was to establish the functional significance of this and to test whether this modulation could interact with stimulation of the NO-cGMP pathway that facilitates the decrease in heart rate (HR) in response to vagal nerve stimulation (VNS). We studied the effect of activation (diazoxide, 100 microM) and inhibition (glibenclamide 30 microM or tolbutamide 5 microM) of K(ATP)channels, and activation of the NO-cGMP pathway with the NO donor, sodium nitroprusside (SNP, 20 microM) or the cGMP analogue, 8-Br-cGMP (0.5 m M) on the HR response to VNS in the isolated guinea pig (Cavia porcellus) double atrial/right vagus preparation (n=40). Tolbutamide increased the bradycardia in response to vagal stimulation at 3 and 5 Hz (P<0.05); effects that were reversed by diazoxide. Glibenclamide also significantly increased the HR response to VNS at 1 and 3 Hz (P<0.05). Diazoxide alone significantly attenuated the HR response to VNS at 5 Hz (P<0.05). Neither glibenclamide nor diazoxide affected the HR response to carbamylcholine (CCh, 50 200 n M). In the presence of a maximal dose of tolbutamide, SNP or 8-Br-cGMP further increased the HR response to VNS at 5 Hz (P<0.05). These results are consistent with the hypothesis that inhibition of sulphonylurea-sensitive channels can increase the HR response to VNS by a pre-synaptic mechanism, and that this modulation may be independent of activation of the NO-cGMP pathway. PMID- 11040110 TI - Impaired cell shortening and relengthening with increased pacing frequency are intrinsic to the senescent mouse cardiomyocyte. AB - Increased heart rate enhances cardiac contractility and accelerates relaxation. Both the force- and relaxation-frequency relationships are critical to myocardial function, especially during stress, and have been shown to be impaired in senescent myocardium. While senescent myocardium is characterized by decreased sarcoplasmic reticulum calcium ATPase activity, it is unclear if altered calcium regulation is directly responsible for the attenuated contractility and relaxation observed with increasing pacing frequency in aged myocardium. We examined this issue using freshly dissociated left ventricular myocytes, isolated from young adult and senescent mouse hearts. Myocytes were paced from 2 to 9 Hz at 37 degrees C, and cell shortening and [Ca(2+)](i)were simultaneously measured using video edge-detection and fura-2 fluorescence, respectively. In adult myocytes, increasing the pacing rate resulted in a progressive increase in percent cell shortening (CS) (P<0.01). This positive CS-frequency relationship was paralleled by an increase in [Ca(2+)](i)transient amplitude (P<0.05). In contrast, the CS-frequency relationship was blunted in senescent myocytes with no increase in percent CS or [Ca(2+)](i)transient amplitude with increasing pacing rate. With increased pacing, the decreases in time constants (tau) of cell relengthening and Ca(2+)transient decay were much steeper in adult compared to senescent myocytes (P<0.05). This study demonstrates that adult mouse myocytes exhibit augmented intracellular Ca(2+)transient amplitude and enhanced intracellular Ca(2+)removal with increasing pacing frequency, resulting in increased cell shortening and enhanced relengthening with frequency. In contrast, senescent mouse myocytes exhibit impaired calcium handling with increasing pacing frequency, which correlated with impairment of both cell shortening and relengthening. PMID- 11040111 TI - Enhanced expression and activity of xanthine oxidoreductase in the failing heart. AB - The molecular basis for heart failure is unknown, but oxidative stress is associated with the pathogenesis of the disease. We tested the hypothesis that the activity of xanthine oxidoreductase (XOR), a free-radical generating enzyme, increases in hypertrophied and failing heart. We studied XOR in two rat models: (1) The monocrotaline-induced right ventricular hypertrophy and failure model; (2) coronary artery ligation induced heart failure, with left ventricular failure and compensatory right ventricular hypertrophy at different stages at 3 and 8 weeks post-infarction, respectively. XOR activity was measured at 30 degrees C and the reaction products were analysed by HPLC. In both models XOR activity in hypertrophic and control ventricles was similar. In the monocrotaline model, the hearts showed enhanced XOR activity in the failing right ventricle (65+/-5 mU/g w/w), as compared to that in the unaffected left ventricle (47+/-3 mU/g P<0.05, n=6-7). In the coronary ligation model, XOR activities did not differ at 3 and 8 weeks. In the infarcted left ventricle, XOR activity increased from 29.4+/-1.4 mU/g (n=6) in sham-operated rats, to 48+/-3 and 80+/-6 mU/g (n=8 P<0.05 v sham) in the viable and infarcted parts of failing rat hearts, respectively. With affinity-purified polyclonal antibody, XOR was localized in CD68+ inflammatory cells of which the number increased more in the failing than in sham-operated hearts. Our results show that the expression of functional XOR is elevated in failing but not in hypertrophic ventricles, suggesting its potential role in the transition from cardiac hypertrophy into failure. PMID- 11040112 TI - Chronic ethanol-induced myocardial protection requires activation of mitochondrial K(ATP) channels. AB - Moderate alcohol consumption protects against coronary heart disease by unclear mechanisms. We tested whether chronic ethanol preconditioning requires activation of mitochondrial K(ATP)channels. Rats were fed 18% (v/v) ethanol in drinking water for 10 months. Blood alcohol levels at sacrifice were 3 mmol/l (0.015 gram percent). Isolated crystalloid-perfused hearts were subjected to global ischemia and reperfusion on a modified Langendorff apparatus. Prior alcohol exposure doubled the recovery of LVDP during reperfusion (45+/-5%v 20+/-3% of baseline for controls, n=6, P<0.01) and blunted the rise in LVEDP (3.5+/-0.5 v 5.5+/-0.4 times baseline for controls, n=6, P<0.01). Ethanol feeding also reduced creatine kinase release during reperfusion. Inhibition of mitochondrial K(ATP)channels with 5 hydroxydecanoate had no effect on baseline LVDP, LVEDP, or coronary flow but abolished the beneficial effects of alcohol on LV contractile recovery and myocyte necrosis. We conclude that mitochondrial K(ATP)channel activity is required for chronic ethanol-induced protection. PMID- 11040114 TI - Beyond receptor expression: the influence of receptor conformation, density, and affinity in HIV-1 infection. PMID- 11040113 TI - A mouse model of vascular injury that induces rapid onset of medial cell apoptosis followed by reproducible neointimal hyperplasia. AB - Genetically modified mice serve as a powerful tool to determine the role of specific molecules in a wide variety of biological phenomena including vascular remodeling. Several models of arterial injury have been proposed to analyze transgenic/knock-out mice, but many questions have been raised about their reproducibility and physiological significance. Here, we report a new mouse model of vascular injury that resembles balloon-angioplasty. A straight spring wire was inserted into the femoral artery via arterioctomy in a small muscular branch. The wire was left in place for one minute to denude and dilate the artery. After the wire was removed, the muscular branch was tied off and the blood flow of the femoral artery was restored. The lumen was enlarged with rapid onset of medial cell apoptosis. While the circumference of the external elastic lamina remained enlarged, the lumen was gradually narrowed by neointimal hyperplasia composed of smooth muscle cells. At 4 weeks, a concentric and homogeneous neointimal lesion was formed reproducibly in the region where the wire had been inserted. Similar exuberant hyperplasia could be induced in all strains examined (C57BL/6J, C3H/HeJ, BALB/c, and 129/SVj). This model may be widely used to study the molecular mechanism of post-angioplasty restenosis at the genetic level. PMID- 11040115 TI - Differential regulation of Bcl-2 and Bax expression in cells infected with virulent and nonvirulent strains of sindbis virus. AB - Sindbis virus is an alphavirus that infects cells in either lytic or persistent infection. In this study we examined the effects of Sindbis virus on cell apoptosis and on the expression of Bcl-2 and Bax. Of the two strains studied, SVA and SVNI, only the neurovirulent strain, SVNI, induced apoptosis of astrocytes and PC-12 cells. SVA, which infects cells in a persistent manner, induced up regulation of bcl-2 mRNA and Bcl-2 protein, whereas SVNI induced an increase in Bax levels. Our results indicate a differential regulation of Bcl2 and Bax expression by SVA and SVNI, which may be associated with the apoptotic potential of the viruses. PMID- 11040116 TI - Effects of echovirus 1 infection on cellular gene expression. AB - To obtain a view of the influence of enterovirus infection on host cell gene expression, multiple cellular mRNA levels were first investigated during echovirus 1 (EV1) infection in HOS palpha2AW cells using cDNA array analysis. Visible cytopathic effect and partial shut-off of host cell protein synthesis were observed 6-10 h after the EV1 infection. Simultaneously, approximately 2% of the investigated genes, among them immediate-early response genes and genes involved in apoptotic pathways and cell growth regulation, were activated over twofold and less than 0.5% of genes were downregulated. For comparison, the cellular effects of coxsackievirus B4 and poliovirus 1 infections were studied in HeLa-Ohio cells by cDNA arrays. Gene activation patterns detected in the host cells during the infection by the three enteroviruses were only partially similar. PMID- 11040117 TI - Ecotropic murine leukemia virus receptor is physically associated with caveolin and membrane rafts. AB - We used a Sindbis virus expression system to stably express a chimeric ecotropic murine leukemia virus (MLV) receptor gene, CAT1, fused to green fluorescent protein (gfp) in BHK cells. The chimeric gene was expressed on the cell surface and functioned as an MLV receptor. Using gfp as an epitope tag, we found that CAT1 cross-immunoprecipitated with caveolin, a cellular protein associated non clathrin-coated endocytic vesicles. Biochemical studies showed that CAT1 copurified with caveolin in a detergent-insoluble membrane fraction that forms cholesterol-rich "rafts" on the cell surface. Disruption of rafts by methyl-beta cyclodextrin, a drug that extracts cholesterol, reduced susceptibility to MLV without decreasing surface CAT1. The results indicate that association of the MLV receptor with cholesterol-rich rafts is important for an early step in virus infection. PMID- 11040118 TI - Use of conventional or replicating nucleic acid-based vaccines and recombinant Semliki forest virus-derived particles for the induction of immune responses against hepatitis C virus core and E2 antigens. AB - Replicating and nonreplicating nucleic acid-based vaccines as well as Semliki Forest-recombinant Viruses (rSFVs) were evaluated for the development of a vaccine against hepatitis C virus (HCV). Replicating SFV-DNA vaccines (pSFV) and rSFVs expressing HCV core or E2 antigens were compared with classical CMV-driven plasmids (pCMV) in single or bimodal vaccine protocols. In vitro experiments indicated that all vaccine vectors produced the HCV antigens but to different levels depending on the antigen expressed. Both replicating and nonreplicating core-expressing plasmids induced, upon injection in mice, specific comparable CTL responses ranging from 10 to 50% lysis (E:T ratio 100:1). Comparison of different injection modes (intramuscular versus intraepidermal) and the use of descalating doses of DNA (1-100 microgram) did not show an increased efficacy of the core-SFV plasmid compared with the CMV-driven one. Surprisingly, rSFVs yielded either no detectable anticore CTL or very low anti-E2 antibody titers following either single or bimodal administration together with CMV-expressing counterparts. Prime boost experiments revealed, in all cases, the superiority of DNA-based only vaccines. The anti-E2 antibody response was evaluated using three different assays which indicated that all generated anti-E2 antibodies were targeted at similar determinants. This study emphasizes the potential of DNA-based vaccines for induction of anti-HCV immune responses and reveals an unexpected and limited benefit of SFV-based vaccinal approaches in the case of HCV core and E2. PMID- 11040119 TI - Inhibition of CCR5-dependent HIV-1 infection by hairpin ribozyme gene therapy against CC-chemokine receptor 5. AB - CCR-5 is a major cellular coreceptor for R5 strains of HIV-1. Individuals carrying a homozygous 32-base-pair deletion in this gene are apparently healthy and are relatively resistant to HIV-1 infection. Since CCR5 appears to be dispensable for the host, but important for initial HIV-1 infection, CCR5 mRNA is an excellent therapeutic target for inhibiting HIV-1 replication via ribozyme knockout. We report here that hairpin ribozymes are able to reduce cellular CCR5 mRNA and cell surface CCR5 when stably introduced into PM1 cells by transduction with recombinant adenoassociated viral vector. The ribozymes effectively protect the cells from infection by R5 HIV-1 strains or non-syncytium-inducing clinical isolates commensurate with a reduction in CCR5 mRNA. These results suggest a novel gene therapy approach to preventing or slowing the disease progression of HIV-1 infection. PMID- 11040120 TI - Partial purification and characterization of Cucumber necrosis virus and Tomato bushy stunt virus RNA-dependent RNA polymerases: similarities and differences in template usage between tombusvirus and carmovirus RNA-dependent RNA polymerases. AB - Tombusviruses are small, plus-sense, single-stranded RNA viruses of plants. RNA dependent RNA polymerases (RdRp) of two tombusviruses, Tomato bushy stunt virus (TBSV) and Cucumber necrosis virus (CNV), have been partially purified from infected Nicotiana benthamiana plants. The obtained RdRp complexes are capable of de novo initiation of complementary RNA synthesis using either plus- or minus strand templates derived from tombusvirus defective interfering (DI) RNAs. In addition to template-sized products, shorter than full-length products were also generated efficiently apparently because of internal initiation of RNA synthesis by the tombusvirus RdRp. This property could be important for the formation of DI RNAs that are observed in tombusvirus infections. The tombusvirus RdRp is also able to use heterologous RNAs derived from satellite RNAs associated with Turnip crinkle virus (TCV) as templates. Generation of full-length, complementary RNA by the tombusvirus RdRp suggests that it can correctly and efficiently recognize the heterologous TCV-specific promoters. Reduced generation of a 3'-terminal extension product in the preceding assay suggests that the previously characterized replication enhancer present in sat-RNA C (Nagy et al., 1999, EMBO J. 18, 5653-5665) does not stimulate tombusvirus RdRp activity. Taken together, these results suggest that template usage by the tombusvirus and carmovirus RdRps are similar, but not identical. PMID- 11040121 TI - Assembly of Sendai virus: M protein interacts with F and HN proteins and with the cytoplasmic tail and transmembrane domain of F protein. AB - Sendai virus matrix protein (M protein) is critically important for virus assembly and budding and is presumed to interact with viral glycoproteins on the outer side and viral nucleocapsid on the inner side. However, since M protein alone binds to lipid membranes, it has been difficult to demonstrate the specific interaction of M protein with HN or F protein, the Sendai viral glycoproteins. Using Triton X-100 (TX-100) detergent treatment of membrane fractions and flotation in sucrose gradients, we report that the membrane-bound M protein expressed alone or coexpressed with heterologous glycoprotein (influenza virus HA) was totally TX-100 soluble but the membrane-bound M protein coexpressed with HN or F protein either individually or together was predominantly detergent resistant and floated to the top of the density gradient. Furthermore, both the cytoplasmic tail and the transmembrane domain of F protein facilitated binding of M protein to detergent-resistant membranes. Analysis of the membrane association of M protein in the early and late phases of the Sendai virus infectious cycle revealed that the interaction of M protein with mature glycoproteins that associated with the detergent-resistant lipid rafts was responsible for the detergent resistance of the membrane-bound M protein. Immunofluorescence analysis by confocal microscopy also demonstrated that in Sendai virus-infected cells, a fraction of M protein colocalized with F and HN proteins and that some M protein also became associated with the F and HN proteins while they were in transit to the plasma membrane via the exocytic pathway. These studies indicate that F and HN interact with M protein in the absence of any other viral proteins and that F associates with M protein via its cytoplasmic tail and transmembrane domain. PMID- 11040122 TI - Interaction of the papillomavirus transcription/replication factor, E2, and the viral capsid protein, L2. AB - The minor capsid protein L2 of papillomaviruses (PVs) likely plays a role in the selective encapsidation of PV DNA in viral capsids and in the infectivity of PV virions. The L2 protein also can cause the relocalization of the PV early protein, E2TA, to nuclear subdomains known as promyelocytic leukemia oncogenic domains (PODs) in which it is localized. E2TA is a transcriptional transactivator that also plays a critical role in viral DNA replication. In this study, we investigated whether L2, in causing the relocalization of E2TA, alters the activities of E2TA. We provide evidence that L2 inhibits the transcriptional transactivation function of E2, but it does not specifically inhibit the capacity of E2 to support viral DNA replication. We also investigated whether the colocalization of E2 and L2 to PODs and the ability of L2 to inhibit the transcriptional transactivation activity of E2TA might be mediated through a direct interaction between these two proteins. Using an in vitro protein-protein association assay, we found that L2 binds to E2TA. Two regions in E2TA were found to mediate this interaction. One of those domains is present in an alternative E2 gene product, E2TR, which is an antagonist to E2TA. Here we show that the L2 protein also relocalizes the E2 transcriptional repressor, E2TR, to the nuclear subdomains. These data suggest that the ability of L2 to relocalize E2 proteins to PODs is mediated through a direct interaction with L2. PMID- 11040123 TI - Mutational analysis of two structural genes of the temperate lactococcal bacteriophage TP901-1 involved in tail length determination and baseplate assembly. AB - Two putative structural genes, orf tmp (tape measure protein) and orf bpp (baseplate protein), of the temperate lactococcal phage TP901-1 were examined by introduction of specific mutations in the prophage strain Lactococcus lactic ssp. cremoris 901-1. The adsorption efficiencies of the mutated phages to the indicator strain L. lactic ssp. cremoris 3107 were determined and electron micrographs were obtained. Specific mutations in orf tmp resulted in the production of mostly phage head structures without tails and a few wild-type looking phages. Furthermore, construction of an inframe deletion or duplication of 29% in orf tmp was shown to shorten or lengthen the phage tail by approximately 30%, respectively. The orf tmp is proposed to function as a tape measure protein, TMP, important for assembly of the TP901-1 phage tail and involved in tail length determination. Specific mutations in orf bpp produced phages which were unable to adsorb to the indicator strain and electron microscopy revealed particles lacking the baseplate structure. The orf bpp is proposed to encode a highly immunogenic structural baseplate protein, BPP, important for assembly of the baseplate. Finally, an assembly pathway of the TP901-1 tail and baseplate structure is presented. PMID- 11040124 TI - Two amino acid substitutions in the SIV Nef protein mediate associations with distinct cellular kinases. AB - A functional Nef protein is crucial in vivo for viral replication leading to pathogenesis in SIV-infected macaques. Moreover, a full-length Nef protein is required for optimal virus replication in primary cells, and both HIV and SIV Nef proteins enhance virion infectivity. Enhanced infectivity may result in part from the ability of Nef to incorporate cellular kinases into virions. In two previous reports, we compared in vitro kinase profiles of SIV recombinant clones that express nef genes derived either from the prototypic lymphocyte-tropic SIVmac239, clone SIV/Fr-2, or from our neurovirulent clone SIV/17E-Fr. While the SIV/Fr-2 Nef protein associated with the previously described PAK-related kinase and an unidentified serine kinase present in a Nef-associated kinase complex (NAKC), SIV/17E-Fr Nef was found to associate with a novel serine kinase activity that was biochemically distinct from both PAK and NAKC. Interestingly, while both Nef proteins were incorporated into virus particles, Nef-associated kinase activity was detected only in virions containing the SIV/17E-Fr Nef protein. Because sequence analysis identified only five amino acids that differed between the Nef proteins of SIV/Fr-2 and SIV/17E-Fr, we were able to evaluate the contribution of each amino acid to Nef-associated kinase activity as well as virus infectivity by constructing a panel of SIV clones containing individual reversions of each differing amino acid in SIV/17E-Fr Nef to the corresponding amino acid in SIV/Fr 2 Nef. In this report, we identify previously uncharacterized amino acids in the N terminus and the conserved core domain of Nef that are essential for the detection of Nef/kinase interactions as well as Nef phosphorylation during SIV infection. Further, via a novel infectivity assay recently developed in our laboratory that utilizes CEMX174 reporter cells stably expressing an SIV/LTR luciferase construct, we find no direct correlation between specific Nef kinase associations and enhanced virion infectivity. PMID- 11040125 TI - Activation of the Lck tyrosine protein kinase by the Herpesvirus saimiri tip protein involves two binding interactions. AB - The Tip protein of Herpesvirus saimiri strain 484C binds to and activates the Lck tyrosine protein kinase. Two sequences in the Tip protein were previously shown to be involved in binding to Lck. A proline-rich region, residues 132-141, binds to the SH3 domain of the Lck protein. We show here that the other Lck-binding domain, residues 104-113, binds to the carboxyl-terminal half of Lck and that this binding does not require the Lck SH3 domain. Mutated Tip containing only one functional Lck-binding domain can bind stably to Lck, although not as strongly as wild-type Tip. Interaction of Tip with Lck through either Lck-binding domain increases the activity of Lck in vivo. Simultaneous binding of both domains is required for maximal activation of Lck. The transient expression of Tip in T cells was found to stimulate both Stat3-dependent and NF-AT-dependent transcription. Mutant forms of Tip lacking one or the other of the two Lck binding domains retained the ability to stimulate Stat3-dependent transcription. Tip lacking the proline-rich Lck-binding domain exhibited almost wild-type activity in this assay. In contrast, ablation of either Lck-binding domain abolished the ability of Tip to stimulate NF-AT-dependent transcription. Full biological activity of Tip, therefore, appears to require both Lck-binding domains. PMID- 11040126 TI - Rabbit hemorrhagic disease virus: genome organization and polyprotein processing of a calicivirus studied after transient expression of cDNA constructs. AB - Rabbit hemorrhagic disease virus (RHDV) belongs to the family Caliciviridae. Studies on this virus are hampered by the lack of a convenient cell culture system. To study viral protein expression a cDNA construct containing the entire protein-coding region of the virus was established and used for transient expression studies. After metabolic labeling of transfected cells and immunoprecipitation with a set of RHDV-specific antisera a variety of polypeptides were identified and assigned to defined regions of the viral genome. The consensus sequences of already identified or putative proteolytic cleavage sites in the viral polyprotein were changed by the introduction of mutations into the expression construct. Expression of these mutated constructs and analysis of the protein patterns allowed us to identify novel cleavage sites in the polyprotein and revealed the first details regarding the order of polyprotein processing. PMID- 11040127 TI - Puumala (PUU) hantavirus strain differences and insertion positions in the hepatitis B virus core antigen influence B-cell immunogenicity and protective potential of core-derived particles. AB - Hepatitis B virus (HBV) core-derived chimeric particles carrying a Puumala (PUU) hantavirus (strain Vranica/Hallnas) nucleocapsid (N) protein sequence (aa 1-45), alternatively inserted at three distinct positions (N-, C-terminus, or the internal region), and mosaic particles consisting of HBV core as well as core/PUU (Vranica/Hallnas) N (aa 1-45) readthrough protein were generated. Chimeric particles carrying the insert at the N-terminus or the internal region of core induced some protective immune response in bank voles (Clethrionomys glareolus) against a subsequent PUU virus (strain Kazan) challenge; 40-50% of the animals showed markers of protection. In contrast, internal insertion of PUU strain CG18 20 N (aa 1-45) into the HBV core caused a highly protective immune response in the bank vole model. Immunizations with particles carrying aa 75-119 of PUU (CG18 20) N at the C-terminus of core verified the presence of a second, minor protective region in the N protein. A strong PUU N-specific antibody response was detected not only in bank voles immunized with chimeric particles containing internal and N-terminal fusions of PUU N protein but also in animals immunized with the corresponding mosaic particles. Except for the exclusive occurrence of antibodies directed against aa 231-240 of N in non-protected animals post virus challenge, there was no additional obvious difference in the epitope-specificity of N-specific antibodies from immunized animals prior and post virus challenge. PMID- 11040128 TI - The structural protein E of the archaeal virus phiCh1: evidence for processing in Natrialba magadii during virus maturation. AB - phiCh1 is a lysogenic virus for the haloalkalophilic archaeon Natrialba magadii. The virus morphology resembles other members of Myoviridae infecting Halobacterium species. The gene of the major capsid protein E of virus phiCh1 was cloned and the DNA sequence was determined. Gene E was mapped to a 3.2-kbp ClaI fragment, localized to the 5'-end of the phiCh1 genome. The complete nucleotide sequence of this region was determined and the identity of gene E was confirmed by comparing the experimentally determined N-terminal amino acid sequence of the purified protein to the translated DNA sequence of its open reading frame. We present evidence that the gene E product is proteolytically cleaved between Lys(16) and Asn(17) to yield the 305 residue polypeptides found in the mature viral capsid. Processing of the protein itself during virus development was determined by 2D gel electrophoresis using protein E-specific antibodies. Sequence similarity studies revealed an 80% identity to capsid protein Hp32 of phiH, infecting Halobacterium salinarum. RT-PCR analysis as well as Western blot studies revealed gene E as a late gene. Transcripts and proteins could be detected shortly before onset of lysis of the lysogenic strain N. magadii L11. PMID- 11040129 TI - Differential phosphorylation of the HPV-16 E7 oncoprotein during the cell cycle. AB - The human papillomavirus type 16 encodes two principal oncoproteins, E6 and E7. The E7 protein has been shown to deregulate the cell cycle through interactions with a variety of proteins involved in cell cycle control and transcriptional regulation. These activities result in E7 being able to cooperate with activated oncogenes in the transformation of primary rodent cells, and with the viral E6 protein during human keratinocyte immortalization. Although a large number of activities have been ascribed to the E7 protein, little is known about its regulation during the cell cycle. We have performed a series of studies to investigate potential changes in E7 phosphorylation during the cell cycle and we show that E7 is indeed differentially phosphorylated. Casein kinase II is the principal kinase during the early part of the cell cycle, but this activity decreases rapidly as cells progress toward S phase. In addition, E7 is transiently phosphorylated at Ser71 in S phase by another, as yet unknown, kinase. These results demonstrate differential regulation of the E7 protein during the cell cycle that most likely represents a means of providing specificity to E7's activities. PMID- 11040130 TI - Cucumber mosaic virus mutants with altered physical properties and defective in aphid vector transmission. AB - Two mutant strains of cucumber mosaic virus (CMV) were investigated with respect to virion stability and molecular determinants of aphid vector transmission. The mutant 2A1-MT-60x, derived from the mechanically passaged wild type 2A1-AT, is poorly transmissible by the aphid Aphis gossypii and not transmissible by the aphid Myzus persicae, whereas the wild type virus is transmissible by both aphid species. The mutant phenotype was shown to be conferred by a single encoded amino acid change of alanine to threonine at position 162 of the coat protein (CP). Modifying the mutant CP gene to encode the wild type sequence (alanine) at position 162 restored aphid transmission. To test for a correspondence between changes in the physical stability of virions and defects in aphid transmission, a urea disruption assay was developed. Virions of aphid-transmissible strains 2A1 AT and CMV-Fny were stable with treatments of up to between 3 and 4 M urea. In this assay mutant viruses 2A1-MT-60x and CMV-M were less stable, as they were completely disrupted at urea concentrations of 2 and 1 M urea, respectively. The mutant 2A1-MT-60x also accumulated at a reduced level in infected squash relative to the wild type virus. These studies suggest that a primary factor in the loss of aphid transmissibility of some strains of CMV is a reduction in virion stability. PMID- 11040131 TI - The GroEL protein of the whitefly Bemisia tabaci interacts with the coat protein of transmissible and nontransmissible begomoviruses in the yeast two-hybrid system. AB - We have previously suggested that a GroEL homolog produced by the whitefly Bemisia tabaci endosymbiotic bacteria interacts in the insect hemolymph with particles of Tomato yellow leaf curl virus from Israel (TYLCV-Is), ensuring the safe circulative transmission of the virus. We have now addressed the question of whether the nontransmissibility of Abutilon mosaic virus from Israel (AbMV-Is) is related to a lack of association between GroEL and the virus coat protein (CP). Translocation analysis has shown that, whereas TYLCV-Is DNA is conspicuous in the digestive tract, hemolymph, and salivary glands of B. tabaci 8 h after acquisition feeding started, AbMV-Is DNA was detected only in the insect digestive tract, even after 96 h. To determine whether AbMV-Is particles were rapidly degraded in the hemolymph as a result of their inability to interact with GroEL, we have isolated a GroEL gene from B. tabaci and used a yeast two-hybrid assay to compare binding of the CP of TYLCV-Is and AbMV-Is to the insect GroEL. The yeast assay showed that the CPs of the two viruses are able to bind efficiently to GroEL. We therefore suggest that, although GroEL-CP interaction in the hemolymph is a necessary condition for circulative transmission, the nontransmissibility of AbMV-Is is not the result of lack of binding to GroEL in the B. tabaci hemolymph, but most likely results from an inability to cross the gut/hemolymph barrier. PMID- 11040132 TI - Influenza A virus neuraminidase: regions of the protein potentially involved in virus-host interactions. AB - Phylogenetically informative amino acid positions (PIPs) were identified in influenza A neuraminidases of subtypes N1 and N2. Neuraminidase evolves in a lineage-specific way as the virus adapts to a new host or changes to evade the host's immune system. Thus, many PIPs undoubtedly identify positions involved in virus-host interactions. Phylogenetically important regions (PIRs) are defined as several PIPs near one another. There are 15 PIRs on N1 and 12 on N2, seven of which are shared between the two subtypes. Many PIRs are coincident with antigenic or glycosylation sites. Other PIRs may represent additional antigenic sites or may be involved in other aspects of virus-host biology. PMID- 11040133 TI - Pseudosubstrate inhibition of protein kinase PKR by swine pox virus C8L gene product. AB - The interferon-induced protein kinase PKR is activated upon binding double stranded RNA and phosphorylates the translation initiation factor eIF2alpha on Ser-51 to inhibit protein synthesis in virally infected cells. Swinepox virus C8L and vaccinia virus K3L gene products structurally resemble the amino-terminal third of eIF2alpha. We demonstrate that the C8L protein, like the K3L protein, can reverse the toxic effects caused by high level expression of human PKR in yeast cells. In addition, expression of either the K3L or C8L gene product was found to reverse the inhibition of reporter gene translation caused by PKR expression in mammalian cells. The inhibitory function of the K3L and C8L gene products in these assays was found to be critically dependent on residues near the carboxyl-termini of the proteins including a sequence motif shared among eIF2alpha and the C8L and K3L gene products. Thus, despite significant sequence differences both the C8L and K3L proteins function as pseudosubstrate inhibitors of PKR. PMID- 11040134 TI - Expression and coreceptor function of APJ for primate immunodeficiency viruses. AB - APJ is a seven transmembrane domain G-protein-coupled receptor that functions as a coreceptor for some primate immunodeficiency virus strains. The in vivo significance of APJ coreceptor function remains to be elucidated, however, due to the lack of an antibody that can be used to assess APJ expression, and because of the absence of an antibody or ligand that can block APJ coreceptor activity. Therefore, we produced a specific monoclonal antibody (MAb 856) to APJ and found that it detected this receptor in FACS, immunofluorescence, and immunohistochemistry studies. MAb 856 also recognized APJ by Western blot, enabling us to determine that APJ is N-glycosylated. Using this antibody, we correlated APJ expression with coreceptor activity and found that APJ had coreceptor function even at low levels of expression. However, we found that APJ could not be detected by FACS analysis on cell lines commonly used to propagate primate lentiviruses, nor was it expressed on human PBMC cultured under a variety of conditions. We also found that some viral envelope proteins could mediate fusion with APJ-positive, CD4-negative cells, provided that CD4 was added in trans. These findings indicate that in some situations APJ use could render primary cell types susceptible to virus infection, although we have not found any evidence that this occurs. Finally, the peptide ligand for APJ, apelin-13, efficiently blocked APJ coreceptor activity. PMID- 11040135 TI - Structure of simian virus 40 DNA replicated by herpes simplex virus type 1. AB - Replicating herpes simplex virus type 1 (HSV-1) DNA is known to form large branched structures. The aim of this study was to define whether HSV-1-specific DNA elements in cis play a critical role in formation of this structure. We did this by investigating the structure of heterologous simian virus 40 (SV40) DNA, which is replicated in HSV-infected cells by SV40 large T-antigen and defined HSV encoded replication factors (e.g., DNA polymerase, single-stranded DNA-binding protein, and helicase-primase). During this process, extrachromosomal concatemeric DNA replication products are formed, indicating a herpesvirus specific replication mode. In this study, we found that the replicating SV40 DNA consisted of a complex branched structure indistinguishable from that of replicating HSV DNA. Thus, no HSV-specific DNA element is necessary in cis for the formation of the large branched structure during HSV DNA replication. The trans-acting HSV DNA replication proteins seem to be sufficient to generate these complex structures. Moreover, replicating SV40 DNA showed a high frequency of homologous recombination events, which is typical for HSV DNA replication. However, in contrast to HSV origin-bearing amplicon plasmids, SV40 plasmids bearing the HSV cleavage-packaging signal were not efficiently processed to linear 150-kb DNA packaged into HSV capsids. This indicates that initiation of DNA synthesis on HSV-ori determines some, yet undefined, property of replicating HSV DNA, which is crucial for regular processing of the replication intermediates to daughter genomes. PMID- 11040137 TI - Accessing electronic information for clinical decisions. PMID- 11040136 TI - Interaction of CD82 tetraspanin proteins with HTLV-1 envelope glycoproteins inhibits cell-to-cell fusion and virus transmission. AB - The entry of retroviruses into their target cell involves interactions between the virus envelope glycoproteins and their cellular receptors, as well as accessory ligand-receptor interactions involving adhesion molecules that can also participate in fusion. We have studied the contribution of CD82 proteins to the transmission of the human T-cell leukemia virus type 1 (HTLV-1), which is greatly dependent on cell-to-cell contacts. CD82 proteins belong to a class of cell surface molecules, the tetraspanins, that can act as molecular facilitators in cellular adhesion processes. The coexpression of CD82 proteins with HTLV-1 envelope glycoproteins resulted in marked inhibition of syncytium formation, whereas CD82 proteins had no effect on syncytium formation induced by human immunodeficiency virus type 1 (HIV-1) envelope proteins. The presence of CD82 proteins also inhibited cell-to-cell transmission of HTLV-1. Coimmunoprecipitation and cocapping experiments showed that CD82 associates with HTLV-1 envelope glycoproteins, both within the cell and at the cell surface. Finally, whereas the intracellular maturation of HTLV-1 glycoproteins was not modified by the presence of CD82 proteins, HTLV-1 protein coproduction delayed the intracellular maturation of CD82 proteins. There thus seems to be a reciprocal interaction between virus and cell proteins, and the cellular proteins involved in adhesion modulate retrovirus transmission both positively, as shown in other systems, and negatively, as shown here. PMID- 11040138 TI - Stamps in paediatrics: Hospitals and clinics. PMID- 11040139 TI - The Anglia Public Health Fellowship--an innovative training opportunity. PMID- 11040140 TI - Neonatal screening for hearing impairment. PMID- 11040142 TI - Fat's not funny PMID- 11040141 TI - Understanding the needs of young asylum seekers. PMID- 11040143 TI - Predictors of somatic symptoms: a five year follow up of adolescents. AB - BACKGROUND: Somatisation is common among adolescents. AIMS: To study factors predicting somatisation later in adulthood. METHODS: Self report questionnaires were administered at baseline examination in 1990 to students (mean age 16.8 years) in schools, and by mail five years later. Results are based on the 615 subjects with no serious disease or injury at baseline. RESULTS: Regression analyses showed that in men the level of somatic symptoms in 1995 was significantly predicted by the respective level in 1990 and by relief smoking. In women, the level of somatic symptoms in 1995 was significantly predicted by the respective level in 1990, self esteem, and the number of negative life events in 1990. After exclusion of cases with a long standing disease in 1995, the multivariate results remained materially similar except that self esteem was no longer significant among women. CONCLUSION: These findings may help in early identification of adolescents with somatisation persisting into early adulthood. PMID- 11040144 TI - A study of clinical opinion and practice regarding circumcision. AB - AIM: To establish clinical opinion regarding appropriate indications for circumcision and to examine actual clinical practice. METHODS: A questionnaire was sent to all NHS hospital consultants in the Yorkshire region of the UK identified as having a role to play in the management of boys (under 16 years of age) requiring circumcision. Retrospective data on actual clinical practice during a three month study period were also collected via a simple proforma. RESULTS: Of 153 questionnaires sent, 64 were returned. Responses revealed varying opinions regarding appropriate indications for circumcision within each consultant group, and between paediatricians and surgeons. Surgeons were generally more inclined to recommend circumcision for each of the indications listed in the questionnaire. Analysis of clinical practice revealed that almost two thirds of procedures were carried out for phimosis, and nearly half of these children were under the age of 5 years. CONCLUSION: There are differences in the clinical opinions of surgeons and paediatricians on what constitutes an appropriate indication for circumcision. Paediatricians' opinions are generally more in line with current evidence than those of surgeons, possibly resulting in many unnecessary circumcisions. PMID- 11040145 TI - Use of the new pneumococcal conjugate vaccine in the USA. PMID- 11040147 TI - Long term effect of abuse PMID- 11040146 TI - Errors in registered birth weight and its implications for mortality statistics. AB - BACKGROUND: Birth weight mortality statistics are important for examining trends and monitoring the outcomes of neonatal care. AIM: To determine the effects of errors in the registered birth weight on birth weight specific mortality. METHODS: All twins born in England and Wales during 1993-95 comprise the denominator population. For those twins that died, the Office for National Statistics (ONS) provided copies of the death certificates. From the information on the death certificates, the registered birth weight was validated and amended using predetermined rules. The neonatal, postneonatal, and infant mortality rates were recalculated. RESULTS: In 2.5% of cases the registered birth weight was "not stated" and in others there were miscoding errors. Important differences between published and amended birth weight specific mortality rates especially in <500 g and >/=3500 g groups were evident. CONCLUSIONS: The bias arising from these errors should be taken into account in interpreting mortality rates and their trends. PMID- 11040148 TI - Imaging in cystic renal disease. PMID- 11040149 TI - Community acquired pneumonia--a prospective UK study. AB - BACKGROUND: There are few data on paediatric community acquired pneumonia (PCAP) in the UK. AIMS: To investigate the aetiology and most useful diagnostic tests for PCAP in the north east of England. METHODS: A prospective study of hospital admissions with a diagnosis of PCAP. RESULTS: A pathogen was isolated from 60% (81/136) of cases, and considered a definite or probable cause of their pneumonia in 51% (70/136). Fifty (37%) had a virus implicated (65% respiratory syncytial virus) and 19 (14%) a bacterium (7% group A streptococcus, 4% Streptococcus pneumoniae), with one mixed infection. Of a subgroup (51 patients) in whom serum antipneumolysin antibody testing was performed, 6% had evidence of pneumococcal infection, and all were under 2 years old. The best diagnostic yield was from paired serology (34%, 31/87), followed by viral immunofluorescence (33%, 32/98). CONCLUSION: Viral infection accounted for 71% of the cases diagnosed. Group A streptococcus was the most common bacterial infective agent, with a low incidence of both Mycoplasma pneumoniae and S pneumoniae. Pneumococcal pneumonia was the most common bacterial cause of pneumonia in children under 2 years but not in older children. Inflammatory markers and chest x ray features did not differentiate viral from bacterial pneumonia; serology and viral immunofluorescence were the most useful diagnostic tests. PMID- 11040150 TI - Streptococcus pneumoniae and Mycoplasma pneumoniae coinfection in community acquired pneumonia. AB - The characteristics of nine children with community acquired pneumonia with evidence of Streptococcus pneumoniae and Mycoplasma pneumoniae coinfection are described. PMID- 11040151 TI - The treatment of convulsive status epilepticus in children. The Status Epilepticus Working Party, Members of the Status Epilepticus Working Party. AB - There is currently little agreement between hospital protocols when treating convulsive status epilepticus in children, and a working party has been set up to produce a national evidence based guideline for treating this condition. This four step guideline is presented in this paper. Its effectiveness will be highlighted and its use audited in a number of centres. PMID- 11040152 TI - Urinary N-acetyl-beta-D-glucosaminidase in epileptic children treated with antiepileptic drugs. AB - AIM: To investigate the effect of prolonged use of antiepileptic drugs on renal function in children. METHODS: Prospective study of 72 children (aged 3-18 years) with epilepsy, on either monotherapy (n = 44) or combined therapy (n = 28). The length of treatment varied from 1 to 13 years. Drugs used were valproic acid, carbamazepine, ethosuximide, clonazepam, clobazepam, and vigabatrin. RESULTS: In 65 patients plasma concentrations of the drugs were in the therapeutic range. In the remaining seven, plasma concentrations were slightly high. In 33 patients urinary N-acetyl-beta-D-glucosaminidase (NAG) activity was raised. The incidence of pathological NAG indices was significantly higher in the combined therapy group than in the monotherapy group. There were also significant differences in the NAG indices of patients depending on the duration of therapy. CONCLUSIONS: Results suggest that chronic use of some antiepileptic drugs-in spite of normal blood concentrations-may alter tubular function, and the dysfunction may result in clinical symptoms. Therefore, we recommend screening of tubular function in these patients. PMID- 11040153 TI - Ventilatory sensitivity to mild asphyxia: prone versus supine sleep position. AB - AIMS: To compare the effects of prone and supine sleep position on the main physiological responses to mild asphyxia: increase in ventilation and arousal. METHODS: Ventilatory and arousal responses to mild asphyxia (hypercapnia/hypoxia) were measured in 53 healthy infants at newborn and 3 months of age, during quiet sleep (QS) and active sleep (AS), and in supine and prone sleep positions. The asphyxial test mimicked face down rebreathing by slowly altering the inspired air: CO(2), maximum 5% and O(2), minimum 13.5%. The change in ventilation with inspired CO(2) was measured over 5-6 minutes of the test. The slope of a linear curve fit relating inspired CO(2) to the logarithm of ventilation was taken as a quantitative measure of ventilatory asphyxial sensitivity (VAS). Sleep state and arousal were determined by behavioural criteria. RESULTS: At 3 months of age, prone positioning in AS lowered VAS (0.184 prone v 0.269 supine, p = 0.050). At newborn age, sleep position had no effect on VAS. Infants aged 3 months were twice as likely to arouse to the test than newborns (p = 0.013). Placing infants prone as opposed to supine increased the chances of arousal 1.57-fold (p = 0.035). CONCLUSION: Our findings show 3 month old babies sleeping prone compared to supine have poorer ventilatory responses to mild asphyxia, particularly in AS, but the increased prevalence of arousal is a protective factor. PMID- 11040154 TI - An association between sudden infant death syndrome (SIDS) and Helicobacter pylori infection. AB - BACKGROUND: Helicobacter pylori has recently been detected in the stomach and trachea of cases of sudden infant death syndrome (SIDS) and proposed as a cause of SIDS. AIMS: To establish the incidence of H pylori in the stomach, trachea, and lung of cases of SIDS and controls. METHODS: Stomach, trachea, and lung tissues from 32 cases of SIDS and eight control cases were examined retrospectively. Diagnosis of SIDS was based on established criteria. Controls were defined by death within 1 year of age and an identifiable cause of death. Tissues were examined histologically for the presence of bacteria. Extracted DNA from these tissues was tested for H pylori ureC and cagA sequences by nested polymerase chain reaction and amplicons detected by enzyme linked immunosorbent assay (ELISA). The cut off for each ELISA for each of the tissue types was taken as the mean optical density plus two times the standard deviation of a range of negative controls. RESULTS: Ages of SIDS cases ranged from 2 to 28 weeks. Ages of controls ranged from 3 to 44 weeks. For the ureC gene, 25 SIDS cases were positive in one or more tissues compared with one of the controls. For the cagA gene, 25 SIDS cases were positive in one or more tissues compared with one of the controls. CONCLUSIONS: There is a highly significant association between H pylori ureC and cagA genes in the stomach, trachea, and lung of cases of SIDS when compared with controls. PMID- 11040155 TI - Late presentation of upper airway obstruction in Pierre Robin sequence. AB - A retrospective review was carried out of 11 consecutive patients with the Pierre Robin sequence referred to a tertiary paediatric referral centre over a five year period from 1993 to 1998. Ten patients were diagnosed with significant upper airway obstruction; seven of these presented late at between 24 and 51 days of age. Failure to thrive occurred in six of these seven infants at the time of presentation, and was a strong indicator of the severity of upper airway obstruction. Growth normalised on treatment of the upper airway obstruction with nasopharyngeal tube placement. All children had been reviewed by either an experienced general paediatrician or a neonatologist in the first week of life, suggesting that clinical signs alone are insufficient to alert the physician to the degree of upper airway obstruction or that obstruction developed gradually after discharge home. The use of polysomnography greatly improved the diagnostic accuracy in assessing the severity of upper airway obstruction and monitoring the response to treatment. This report highlights the prevalence of late presentation of upper airway obstruction in the Pierre Robin sequence and emphasises the need for close prospective respiratory monitoring in this condition. Objective measures such as polysomnography should be used, as clinical signs alone may be an inadequate guide to the degree of upper airway obstruction. PMID- 11040156 TI - Paraparesis, hypermanganesaemia, and polycythaemia: a novel presentation of cirrhosis. AB - Progressive myelopathy is a rare complication of chronic hepatic disease which has never been reported in the paediatric age group. We describe the 11 year course of an adolescent male with hepatic myelopathy caused by cryptogenic micronodular cirrhosis. His condition has been associated with persistent polycythaemia and extraordinary increases of whole blood manganese, with magnetic resonance imaging evidence of manganese deposition within the basal ganglia and other regions of the brain. The patient has developed neither liver failure nor parkinsonism. The pathophysiological bases of this multiorgan system disorder are described. PMID- 11040157 TI - Pseudoinfectious mononucleosis: a presentation of Bartonella henselae infection. AB - Six children presented during one year with clinical features of infectious mononucleosis, but with laboratory findings of leucocytosis with neutrophilia, increased erythrocyte sedimentation rate, and hypergammaglobulinaemia. Serology for Epstein-Barr virus, cytomegalovirus, adenovirus, and Toxoplasma gondii was negative, while anti-Bartonella henselae IgM with high IgG titre (>/=1/1024) was present in all. All children had contact with kittens. No specific treatment was administered and all recovered. PMID- 11040158 TI - Recollection of children following intensive care. AB - BACKGROUND AND AIMS: The recollections of critically ill children following discharge from the paediatric intensive care unit (PICU) have not previously been described. We have interviewed such children to establish the nature of their recollections. METHODS: Children aged 4 years and above were interviewed following discharge from the PICU at the Queens Medical Centre, Nottingham, either in hospital or at home, using a semistructured interview. Their recollections were recorded and interpreted by content analysis. RESULTS: A total of 38 interviews were carried out; 44 specific recollections were reported, the majority being neutral (60%) or positive (25%). Only 15% of recollections were negative. Negative recollections related to aspects of medical care and environmental factors. No child treated with neuromuscular blocking agents remembered any period of therapeutic paralysis. CONCLUSIONS: Children's recollections of PICU are mainly neutral or positive. Mechanically ventilated children sedated with midazolam and morphine remember little of endotracheal intubation. PMID- 11040159 TI - Autosomal recessive osteopetrosis: diagnosis, management, and outcome. PMID- 11040160 TI - Assessing the child with scoliosis: the role of surface topography. PMID- 11040161 TI - Assessing outcomes in twin-twin transfusion syndrome. PMID- 11040162 TI - Twin-twin transfusion syndrome: a five year review. AB - OBJECTIVE: To determine the incidence, complications, management, and outcome in infants with twin-twin transfusion syndrome (TTTS) over a period of five years. METHODS: TTTS was diagnosed in monochorionic twins if one was pale and the other plethoric with a haemoglobin difference > or =5 g/100 ml and/or birthweight differences > or =15%. RESULTS: Eighteen (6.2%) of the 292 twin pairs had TTTS. Eight pairs (44%) had the acute type and the rest (56%) had the chronic type of TTTS. The mean (SEM) intrapair haemoglobin difference in the acute type was 4.8 (2.1) g/100 ml which gave a discordance of 7.1 (4.6)%, whereas that in the chronic type was 6.9 (2.9) g/100 ml and 24.4 (6.1)% respectively. Infants with the acute type had a significantly higher incidence of vaginal delivery (p<0.03), hypotension (p<0.025), and respiratory distress (p<0.01) compared with those with the chronic type. There was no significant difference in the incidence of anaemia, polycythaemia, asphyxia, hypoglycaemia, and hyperbilirubinaemia. Two recipients died in utero as the result of chronic TTTS, while their survivors developed spastic cerebral palsy. There were no neonatal deaths. CONCLUSIONS: TTTS, although uncommon, may have an adverse neurodevelopmental outcome especially if one twin dies in utero. Prompt recognition and management of the haemodynamic and haematological problems of infants with the acute types of TTTS will result in optimal neurodevelopmental outcome. PMID- 11040163 TI - Long term outcome of twin-twin transfusion syndrome. AB - AIMS: To compare the perinatal mortality and morbidity of infants with twin-twin transfusion syndrome (TTTS) with those of gestation matched twin controls and to assess the neurodevelopmental outcome of surviving twins with TTTS. METHODS: A cohort of 17 consecutive pregnancies with TTTS was enrolled over three years together with gestation matched twin pregnancies unaffected by TTTS. Serial amnioreduction for the TTTS pregnancies was performed as appropriate. Perinatal death and neonatal morbidities were recorded for both the TTTS cohort and controls. The TTTS survivors had neurodevelopmental follow up to at least 2 years of age. RESULTS: In 12 of the pregnancies, serial amniocenteses were performed, but, in five, the infants were born before intervention. The mean gestational age at delivery was 29.1 weeks (range 23-36). There were five intrauterine deaths in the TTTS cohort and six neonatal deaths (survival 68%). In the control group, there was one intrauterine death and five neonatal deaths (survival 82%). Infants in the TTTS group had a greater requirement for inotropes (p = 0.04) and a higher incidence of renal failure (p = 0.005). Periventricular leucomalacia and cerebral atrophy were seen in 17% of the TTTS group, but none of the controls (p = 0.03). The 23 surviving TTTS infants were all followed up, with 22% having significant neurological morbidity: cerebral palsy and global developmental delay. CONCLUSIONS: Twins with TTTS have high perinatal mortality and neonatal morbidity, and long term neurodevelopmental morbidity in survivors is high. Further investigation into the pathogenesis and management of TTTS is required. PMID- 11040164 TI - Early postnatal dexamethasone treatment and increased incidence of cerebral palsy. AB - OBJECTIVE: To study the long term neurodevelopmental outcome of children who participated in a randomised, double blind, placebo controlled study of early postnatal dexamethasone treatment for prevention of chronic lung disease. METHODS: The original study compared a three day course of dexamethasone (n = 132) with a saline placebo (n = 116) administered from before 12 hours of age in preterm infants, who were ventilated for respiratory distress syndrome and had received surfactant treatment. Dexamethasone treatment was associated with an increased incidence of hypertension, hyperglycaemia, and gastrointestinal haemorrhage and no reduction in either the incidence or severity of chronic lung disease or mortality. A total of 195 infants survived to discharge and five died later. Follow up data were obtained on 159 of 190 survivors at a mean (SD) age of 53 (18) months. RESULTS: No differences were found between the groups in terms of perinatal or neonatal course, antenatal steroid administration, severity of initial disease, or major neonatal morbidity. Dexamethasone treated children had a significantly higher incidence of cerebral palsy than those receiving placebo (39/80 (49%) v. 12/79 (15%) respectively; odds ratio (OR) 4.62, 95% confidence interval (95% CI) 2.38 to 8.98). The most common form of cerebral palsy was spastic diplegia (incidence 22/80 (28%) v. 5/79 (6%) in dexamethasone and placebo treated infants respectively; OR 4.45, 95% CI 1.95 to 10.15). Developmental delay was significantly more common in the dexamethasone treated group (44/80 (55%)) than in the placebo treated group (23/79 (29%); OR 2. 87, 95% CI 1.53 to 5.38). Dexamethasone treated infants had more periventricular leucomalacia and less intraventricular haemorrhage in the neonatal period than those in the placebo group, although these differences were not statistically significant. Eleven children with cerebral palsy had normal ultrasound scans in the neonatal period; all 11 had received dexamethasone. Logistic regression analysis showed both periventricular leucomalacia and drug assignment to dexamethasone to be highly significant predictors of abnormal neurological outcome. CONCLUSIONS: A three day course of dexamethasone administered shortly after birth in preterm infants with respiratory distress syndrome is associated with a significantly increased incidence of cerebral palsy and developmental delay. PMID- 11040165 TI - Sex differences in outcomes of very low birthweight infants: the newborn male disadvantage. AB - OBJECTIVE: To determine the differences in short term outcome of very low birthweight infants attributable to sex. METHODS: Boys and girls weighing 501 1500 g admitted to the 12 centres of the National Institute of Child Health and Human Development Neonatal Research Network were compared. Maternal information and perinatal data were collected from hospital records. Infant outcome was recorded at discharge, at 120 days of age if the infant was still in hospital, or at death. Best obstetric estimate based on the last menstrual period, standard obstetric factors, and ultrasound were used to assign gestational age in completed weeks. Data were collected on a cohort that included 3356 boys and 3382 girls, representing all inborn births from 1 May 1991 to 31 December 1993. RESULTS: Mortality for boys was 22% and that for girls 15%. The prenatal and perinatal data indicate few differences between the sex groups, except that boys were less likely to have been exposed to antenatal steroids (odds ratio (OR) = 0.80) and were less stable after birth, as reflected in a higher percentage with lower Apgar scores at one and five minutes and the need for physical and pharmacological assistance. In particular, boys were more likely to have been intubated (OR = 1.16) and to have received resuscitation medication (OR = 1.40). Boys had a higher risk (OR > 1.00) for most adverse neonatal outcomes. Although pulmonary morbidity predominated, intracranial haemorrhage and urinary tract infection were also more common. CONCLUSIONS: Relative differences in short term morbidity and mortality persist between the sexes. PMID- 11040166 TI - Importance of intestinal colonisation in the maturation of humoral immunity in early infancy: a prospective follow up study of healthy infants aged 0-6 months. AB - AIM: To evaluate the role of intestinal microflora and early formula feeding in the maturation of humoral immunity in healthy newborn infants. STUDY DESIGN: Sixty four healthy infants were studied. Faecal colonisation with Bacteroides fragilis group, Bifidobacterium-like, and Lactobacillus-like bacteria was examined at 1, 2, and 6 months of age, and also the number of IgA-secreting, IgM secreting, and IgG-secreting cells (detected by ELISPOT) at 0, 2, and 6 months of age. RESULTS: Intestinal colonisation with bacteria from the B fragilis group was more closely associated with maturation of IgA-secreting and IgM-secreting cells than colonisation with the other bacterial genera studied or diet. Infants colonised with B fragilis at 1 month of age had more IgA-secreting and IgM secreting cells/10(6) mononuclear cells at 2 months of age (geometric mean (95% confidence interval) 1393 (962 to 2018) and 754 (427 to 1332) respectively) than infants not colonised (1015 (826 to 1247) and 394 (304 to 511) respectively); p = 0.04 and p = 0.009 respectively. CONCLUSIONS: The type of bacteria colonising the intestine of newborns and the timing may determine the immunomodulation of the naive immune system. PMID- 11040167 TI - Leptin and metabolic hormones in infants of diabetic mothers. AB - AIMS: To investigate the effect of maternal diabetes on leptin in term newborns and to determine whether leptin correlates with insulin and its associated biochemical parameters in support of the hypothesis that a functional "adipoinsular axis" might exist at this stage of development. METHODS: A total of 116 term newborns were prospectively enrolled and categorised into three groups: 44 were infants of non-diabetic mothers (control group C); 41 were infants born to mothers with gestational diabetes on dietary treatment (group D); and 31 were infants born to mothers with gestational or pregestational diabetes on insulin treatment (group I). RESULTS: No significant difference in serum leptin was observed between the three groups; the results of the study population were therefore pooled and analysed. Serum leptin correlated significantly with serum insulin, insulin:glucose ratio, birth weight, body length, body mass index, placenta weight, and maternal HbA(1c). Female infants had significantly higher serum leptin than male infants. All parameters except placenta weight and body length remained significantly associated with serum leptin when multivariate stepwise regression analysis was applied. Subgroup analysis revealed a significant correlation between serum leptin and cortisol in group D. CONCLUSIONS: There was no significant difference in serum leptin between infants born to diabetic and non-diabetic mothers, though infants born to mothers requiring insulin treatment had the highest median serum leptin concentrations. The significant association between serum leptin and insulin or insulin:glucose ratio supports the hypothesis that a functional adipoinsular axis might exist in term newborns. Furthermore, the significant correlation between maternal HbA(1c) and circulating leptin of the studied infants suggests that the clinical control of maternal diabetes could affect the regulation of serum leptin in these infants. PMID- 11040168 TI - Leptin and metabolic hormones in preterm newborns. AB - AIM: To investigate the inter-relation between leptin and other metabolic hormones in preterm and term infants and to explore whether a functional "adipoinsular axis" might exist in preterm newborns. METHODS: A total of 140 preterm and term newborns were prospectively recruited and categorised according to gestation length. Blood samples were taken at 24 hours (day 1), and on day 4-5 of life. RESULTS: Serum leptin, cortisol, free thyroxine, and plasma ACTH on day 1 were significantly higher in term than in preterm infants. The relation between serum leptin and gestation followed a non-linear pattern; the slope of the curve began to increase steeply between 33 and 35 weeks gestation. Serum leptin on day 1 was significantly associated with serum insulin, insulin:glucose ratio, and plasma ACTH in infants less than 34 weeks gestation; serum leptin on day 1 and day 4-5 were significantly correlated with insulin:glucose ratio in infants 34 or more weeks gestation. Significant changes in the pattern of metabolic hormones were observed in the first week of life. Serum insulin and plasma glucose were significantly increased between day 1 and day 4-5; serum leptin was significantly decreased. CONCLUSIONS: The circulating leptin concentration increases markedly after 34 weeks gestation and bears a close temporal relation with the exponential accumulation of body fat mass during that period. The inter-relation between serum leptin and insulin or insulin:glucose ratio before and after 34 weeks gestation indicates that the "adipoinsular axis" is likely to be functional in early (<34 weeks gestation) intrauterine life. The rapid decline in the circulating concentrations of leptin after birth may be of physiological advantage to preterm and term newborns by limiting their body energy expenditure and conserving nutritional reverses for subsequent growth and development. PMID- 11040169 TI - Resonance frequency in respiratory distress syndrome. AB - AIM: To observe how the resonance frequency changes with the course of respiratory distress syndrome (RDS), by examining the effect of changing static compliance on the resonance frequency in premature infants. METHODS: In 12 ventilated premature infants with RDS (mean gestational age 26.6 weeks, mean birth weight 0.84 kg), resonance frequency and static compliance were determined serially using phase analysis and single breath mechanics technique respectively in the first seven days of life. RESULTS: The minimum number of measurements done in any one baby was three and maximum was five in this seven day study period. The first measurement in each baby was done within the first 72 hours of life. The increase in compliance in this period varied from 27% to 179%. The variation in the corresponding resonance frequency was within 2 Hz in eight babies and within 6 Hz in all recruited babies. CONCLUSIONS: The resonance frequency of the respiratory system in preterm infants with RDS remains remarkably constant in the early stages of the illness, despite relatively large changes in static compliance. PMID- 11040170 TI - Does topical amethocaine gel reduce the pain of venepuncture in newborn infants? A randomised double blind controlled trial. AB - BACKGROUND: Topical amethocaine provides effective pain relief during venepuncture in children, and has been shown to have a local anaesthetic action in the newborn. AIM: To investigate the effect of topical amethocaine on the pain of venepuncture in the newborn. DESIGN: Randomised double blind placebo controlled trial. SUBJECTS: Forty newborn infants, gestation 27-41 weeks (median 33), age 2-17 days (median 7), undergoing routine venepuncture. METHOD: A 1.5 g portion of 4% (w/w) amethocaine gel (Ametop; Smith and Nephew, Hull, UK) or placebo was applied to the skin under occlusion for one hour, then wiped away. Venepuncture was performed five minutes later. Facial reaction and cry were recorded on videotape. Pain was assessed using a validated adaptation of the neonatal facial coding system. Five features were scored at one second intervals for five seconds before and after venepuncture. No or minimal pain was defined as a cumulative score of below 10 (out of 25) in the five seconds after needle insertion. Each author scored the tapes independently. RESULTS: There was close agreement on scoring of the tapes. One infant was excluded because of restlessness before the venepuncture. Of 19 amethocaine treated infants, 16 (84%) showed little or no pain compared with six of 20 (30%) in the placebo group (p = 0.001). The median cumulative neonatal facial coding system score over five seconds after needle insertion was 3 compared with 16 in the placebo group (p = 0.001). Of the 19 amethocaine treated infants, 15 (79%) did not cry compared with five of 20 (25%) placebo treated infants (p = 0.001). No local reaction to amethocaine was seen. CONCLUSION: Topical amethocaine provides effective pain relief during venepuncture in the newborn. PMID- 11040171 TI - Topical amethocaine gel in the newborn infant: how soon does it work and how long does it last? AB - AIM: To explore the time of onset and duration of action of topical amethocaine gel in the newborn infant. DESIGN: A randomised double blind placebo controlled trial. SUBJECTS: Thirty six infants were studied after 30 minutes application and 36 after 60 minutes application. A total of 56 infants (gestation 27-42 weeks, weight 0. 79-4.1 kg) were studied in the first two weeks after delivery. METHOD: 1.5 g amethocaine or placebo was applied to the dorsum of either foot, occluded, and then left for 30 or 60 minutes. Local anaesthesia was assessed by observing the cutaneous withdrawal response to graded nylon filaments (von Frey hairs). If there was a difference between feet in filament thickness required to elicit a response, the infant was studied in an identical manner at hourly intervals until the difference had disappeared. RESULTS: Evidence of local anaesthetic action of amethocaine was seen in 23 of 36 (64%) infants after 30 minutes and 26 of 36 (72%) infants after 60 minutes application (no significant difference). Evidence of local anaesthetic action was independent of gestation and order of testing. Amethocaine responders showed a significantly deeper anaesthetic action than placebo responders. The median duration of action was 1.5 hours (range 0.5-3.5) after 30 minutes and three hours (range 1-5) after 60 minutes (p<0.001). CONCLUSION: Topical amethocaine gel has a local anaesthetic action after 30 minutes application, but application for 60 minutes results in longer duration of action. PMID- 11040172 TI - Feeding issues in preterm infants. PMID- 11040173 TI - Early enteral feeding of the preterm infant. PMID- 11040174 TI - Dr Emmett Holt (1855-1924) and the foundation of North American paediatrics. PMID- 11040175 TI - Winning the war and losing the peace? PMID- 11040176 TI - A multicenter trial of 6-mercaptopurine and prednisone in children with newly diagnosed Crohn's disease. AB - BACKGROUND & AIMS: Clinical experience suggests that 6-mercaptopurine (6-MP) is effective therapy for children with active steroid-dependent Crohn's disease (CD). We report the results of a prospective, placebo-controlled, multicenter trial evaluating the combination of 6-MP and prednisone as therapy for children with newly diagnosed moderate-to-severe CD. METHODS: Fifty-five children (age, 13+/-2 years) were randomized to treatment with 6-MP (1.5 mg x kg(-1) x day(-1)) or placebo within 8 weeks of initial diagnosis. Both groups also received prednisone (40 mg/day). Prednisone dosage adjustments were based on a defined schedule determined by the change in a subject's disease activity score, and steroid administration was discontinued as remission was achieved. Study treatment with 6-MP or placebo continued for 18 months. RESULTS: Groups were comparable for age, sex, and site and activity of disease. In the 6-MP group, the duration of steroid use was shorter (P<0.001) and the cumulative steroid dose lower at 6, 12, and 18 months (P<0.01). Although remission was induced in 89% of both groups, only 9% of the remitters in the 6-MP group relapsed compared with 47% of controls (P = 0.007). Growth was comparable in both groups. No clinically significant adverse events occurred, although mild leukopenia and increases in aminotransferase activity were noted in the 6-MP group. CONCLUSIONS: Addition of 6-MP to a regimen of corticosteroids significantly lessens the need for prednisone and improves maintenance of remission. 6-MP should be part of the initial treatment regimen for children with newly diagnosed moderate-to-severe CD. PMID- 11040177 TI - Comparison of heparin and steroids in the treatment of moderate and severe ulcerative colitis. AB - BACKGROUND & AIMS: Unfractionated heparin has been found to reduce symptoms and improve healing as adjuvant therapy in patients with ulcerative colitis. The current study evaluated the efficacy and safety of unfractionated heparin in the treatment of ulcerative colitis in comparison with methylprednisolone. METHODS: A multicenter randomized trial with blinded endpoint evaluation was conducted in patients hospitalized for moderate or severe ulcerative colitis. Patients were randomized to receive heparin as a continuous infusion or methylprednisolone (0.75-1 mg x kg(-1) x day(-1)). RESULTS: Twenty-five patients entered the study: 13 received methylprednisolone and 12 received heparin. By day 10, 69% of patients in the methylprednisolone group, but none in the heparin group, achieved significant improvement or remission. C-reactive protein levels significantly decreased in the methylprednisolone group but not in the heparin group. Three patients in the heparin group were withdrawn before day 10 because of an adverse event: rectal bleeding needing transfusion (2 cases) or surgery (1 case). The proportion of patients with persistent rectal bleeding at day 10 was 31% in the methylprednisolone group and 90% in the heparin group (P<0.05). CONCLUSIONS: Unfractionated heparin as monotherapy is not effective in the treatment of moderate or severe ulcerative colitis and is associated with significant bleeding complications. PMID- 11040178 TI - Genetic factors determine extent of bone loss in inflammatory bowel disease. AB - BACKGROUND & AIMS: Although bone loss and osteoporosis are well-known long-term sequelae of inflammatory bowel disease (IBD), the risk factors for increased bone loss have not been identified. Balances of pro- and anti-inflammatory cytokines influence mechanisms of both chronic inflammation and bone resorption. The aim of this study was to identify genetic risk factors for rapid bone loss in IBD patients as a model of disease- and inflammation-associated bone loss. METHODS: Multiple clinical parameters, biochemical markers of bone metabolism (vitamin D, parathyroid hormone, N-terminal telopeptide of type-I collagen, desoxypyridinoline, bone alkaline phosphatase), and bone mineral density were prospectively assessed in 83 IBD patients over 1.6+/-0.3 years. Eighty-six healthy bone marrow donors served as controls for allelotyping. The allele status of the interleukin 1 receptor antagonist (IL-1ra), IL-6, heat shock protein 70-2 (hsp 70-2), and heat shock protein 70-hom (hsp hom) genes was typed and correlated with clinical course of IBD and extent of bone loss. RESULTS: The extent of bone loss was not correlated to clinical severity of disease or application of corticosteroids. Noncarriage of the 240-base pair allele of the IL 1ra gene and carriage of the 130-base pair allele of IL-6 were independently associated with increased bone loss. Genetic variations of the hsp genes were not associated with degree of bone loss. The combined presence of the named risk factors was significantly associated with increasing bone loss. CONCLUSIONS: Genetic variations in the IL-6 and IL-1ra gene identify IBD patients at risk for increased bone loss. PMID- 11040179 TI - Microsatellite instability is a favorable prognostic indicator in patients with colorectal cancer receiving chemotherapy. AB - BACKGROUND & AIMS: Adjuvant 5-fluorouracil (5-FU)-based chemotherapy is standard treatment for stage C colorectal cancer (CRC). Approximately 12% of CRCs are characterized by microsatellite instability (MSI), a hallmark of a DNA mismatch repair defect. We investigated the safety of adjuvant 5-FU-based chemotherapy for MSI(+) CRC and compared the prognosis of MSI(+) and MSI(-) CRC patients receiving adjuvant therapy. METHODS: Previously, a prospective series consisting of 1044 consecutive CRCs has been collected and the MSI status of each sample determined. Patients with stage C cancer who had received adjuvant chemotherapy (n = 95) were followed up for 7-63 months (median, 31 months) after surgery. RESULTS: No unexpected or serious adverse effects were observed when 5-FU-based chemotherapy was used as adjuvant treatment for MSI(+) CRC. Three- year recurrence-free survival was 90% and 43% in the MSI(+) (n = 11) and MSI(-) (n = 84) groups, respectively (P = 0.020). CONCLUSIONS: Adjuvant 5-FU-based chemotherapy is feasible for both MSI(+) and MSI(-) CRCs, and patients with MSI(+) CRC who receive adjuvant therapy have an excellent prognosis. PMID- 11040180 TI - p16(INK4a) expression begins early in human colon neoplasia and correlates inversely with markers of cell proliferation. AB - BACKGROUND & AIMS: p16(INK4a) is a cell cycle inhibitor and a major tumor suppressor protein, but the regulation of p16(INK4a) is poorly understood and the physiologic settings in which it exerts its antiproliferative effects are unknown. A role for p16(INK4a) in intestinal neoplasia is suggested by the observation that the promoter region is methylated in a subset of human colon tumors. We examined the expression of the protein in specimens representing the full spectrum of neoplastic progression in the human colon and determined whether expressing cells showed evidence of cell cycle inhibition. METHODS: We studied p16(INK4a) expression by immunoprecipitation, immunoblotting, reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry, and immunofluorescence in matched normal and neoplastic colonic tissue from 70 patients. RESULTS: p16(INK4a) expression was very low in normal mucosa, with staining observed in rare epithelial cells at the base of crypts. A distinctly higher expression was found in 4 of 7 aberrant crypt foci, 32 of 36 adenomas, 18 of 28 primary carcinomas, and 5 of 5 metastatic carcinomas. Within each neoplasm p16(INK4a) staining was heterogeneous, with higher expression commonly seen in areas bordering normal tissue. p16(INK4a) staining correlated inversely with that of Ki67, cyclin A, and the retinoblastoma protein, suggesting that cell cycle progression was inhibited. CONCLUSIONS: These results suggest that p16(INK4a) expression begins in the earliest detectable stages of neoplastic progression in the human colon and exerts a continuous, piecemeal constraint on tumor growth. PMID- 11040181 TI - The effects of intestinal infusion of long-chain fatty acids on food intake in humans. AB - BACKGROUND & AIMS: Dietary fat intake is related to the degree of obesity, but the specific mechanisms by which fats regulate food intake in humans are unclear. We compared food intake suppression, plasma triglyceride appearance, and cholecystokinin (CCK) response after intestinal infusion of oils enriched with C18 fatty acids of increasing unsaturation. METHODS: Food intake and appetite changes after upper intestinal infusion of 0.9% saline, 20% Intralipid, and 20% emulsions of oils enriched with stearic, oleic, and linoleic acids were tested in 10 healthy male volunteers. Plasma triglyceride appearance and CCK release were tested separately in 7 additional volunteers. RESULTS: Intralipid and linoleic acid infusions significantly reduced food intake compared with saline infusion (P<0.05). No changes were observed in appetite ratings. There were no differences in plasma triglyceride response over the initial 75 minutes of intestinal infusion. Plasma CCK concentration increased after all lipid infusions (P<0.001), Intralipid infusion produced the highest increase in plasma CCK (P<0.05), and CCK response was similar between the 3 enriched oil emulsions. CONCLUSIONS: These results indicate marked differences in the ability of C18 fatty acids to reduce food intake that appear not to be related to rate of absorption but may partially be explained by CCK release. PMID- 11040182 TI - The specific activities of human digestive lipases measured from the in vivo and in vitro lipolysis of test meals. AB - BACKGROUND & AIMS: The lipolytic potential of digestive lipases in vivo has always been deduced so far from their in vitro activities under nonphysiologic conditions. In the present study, the specific activities of human gastric lipase (HGL) and pancreatic lipase (HPL) were measured on dietary triglycerides (TGs) during test meal lipolysis. METHODS: Healthy human volunteers ingested a liquid or solid meal. The specific activities of HGL and HPL were estimated from the lipase and free fatty acid (FFA) outputs at the postpyloric and duodenal levels, respectively. Based on the in vivo data, lipolysis was also performed in vitro by mixing the meal either with gastric juice and subsequently with pancreatic juice and bile or with purified HGL and HPL. FFAs were measured by thin-layer chromatography, and the specific activities of HGL and HPL were expressed as micromoles of FFA per minute per milligram of lipase. RESULTS: In vitro, the specific activities on the liquid meal TGs were 32 (gastric juice) and 34 (pure lipase) micromol x min(-1) x mg(-1) with HGL and 47 (pancreatic juice) and 43 (pure lipase) micromol x min(-1). mg(-1) with HPL. The specific activities on the solid meal TGs were 33 (gastric juice) and 32 (pure lipase) micromol x min(-1) x mg(-1) with HGL and 12 (pancreatic juice) and 15 (pure lipase) micromol x min(-1) x mg(-1) with HPL. The in vivo values obtained were in the same range. The secretory lipase outputs were 21.6+/-14.5 mg HGL and 253.5+/-95.5 mg HPL with the liquid test meal and 15.2+/-5.1 mg HGL and 202.9+/-96.1 mg HPL with the solid test meal. CONCLUSIONS: The specific activities of HGL and HPL on meal TGs were much lower than those measured in vitro under optimized assay conditions (1300 8000). However, these low specific activities are enough for the meal TGs to be completely lipolysed, given the amounts of HGL and HPL secreted during a meal. PMID- 11040183 TI - Cdx1 and cdx2 expression during intestinal development. AB - BACKGROUND & AIMS: The intestine-specific transcription factors Cdx1 and Cdx2 are candidate genes for directing intestinal development, differentiation, and maintenance of the intestinal phenotype. This study focused on the complex patterns of expression of Cdx1 and Cdx2 during mouse gastrointestinal development. METHODS: Embryonic and postnatal mouse tissues were analyzed by immunohistochemistry to determine protein expression of Cdx1 and Cdx2 in the developing intestinal tract. RESULTS: Cdx2 protein expression was observed at 9. 5 postcoitum (pc), whereas weak expression of Cdx1 protein was first seen at 12.5 pc in the distal developing intestine (hindgut). Expression of Cdx1 increased from 13.5 to 14.5 pc during the endoderm/epithelial transition with predominately distal expression. In contrast to Cdx1, there was intense expression of Cdx2 in all but the distal portions of the developing intestine. Cdx2 expression remained low in the distal colon throughout postnatal development. A gradient of expression formed in the crypt-villus axis, with Cdx1 primarily in the crypt and Cdx2 primarily in the villus. CONCLUSIONS: Direct comparison of the patterns of Cdx1 and Cdx2 protein expression during development as performed in this study provides new insights into their potential functional roles. The relative expression of Cdx1 to Cdx2 protein may be important in the anterior to posterior patterning of the intestinal epithelium and in defining patterns of proliferation and differentiation along the crypt-villus axis. PMID- 11040184 TI - Interleukin 16 is up-regulated in Crohn's disease and participates in TNBS colitis in mice. AB - BACKGROUND & AIMS: Interleukin (IL)-16 is a T lymphocyte- derived cytokine that uses CD4 as its receptor and hence selectively recruits CD4-bearing cells. Infiltrating CD4(+) T cells are a feature of Crohn's disease; however, the role of IL-16 in intestinal inflammation is unknown. The aim of this study was to determine whether IL-16 production is increased in inflammatory bowel disease and whether IL-16 participates in trinitrobenzene sulfonic acid (TNBS)-induced colitis in mice. METHODS: IL-16 messenger RNA and protein levels in inflammatory bowel disease tissues were determined by reverse-transcription polymerase chain reaction and enzyme-linked immunosorbent assay. C57BL/6 or BALB/c mice were treated with vehicle, TNBS alone, TNBS + anti-IL-16 monoclonal antibody (mAb), TNBS + control mAb, or were untreated. Colonic injury and inflammation were evaluated after 3 or 10 days. RESULTS: Colonic IL-16 protein levels were increased in patients with Crohn's disease (P<0.05) but not ulcerative colitis. Anti-IL-16 mAb treatment significantly reduced TNBS-induced weight loss (P< 0.001), mucosal ulceration (P<0.05), myeloperoxidase activity (P< 0.001), and TNBS-mediated increases in mucosal levels of IL-1beta (P<0.05) and tumor necrosis factor alpha (P<0.01). CONCLUSIONS: Anti-IL-16 mAb reduced colonic injury and inflammation induced by TNBS in mice. Colonic mucosal IL-16 levels were elevated in Crohn's disease, suggesting a role for IL-16 in the pathophysiology of inflammatory bowel disease. PMID- 11040185 TI - Limited CD4 T-cell diversity associated with colitis in T-cell receptor alpha mutant mice requires a T helper 2 environment. AB - BACKGROUND & AIMS: T-cell receptor alpha mutant (TCRalpha(-/-)) mice spontaneously develop chronic colitis mediated by CD4(+) TCRalpha(-)beta(+) T cells. The aim of this study was to analyze the mechanisms of expansion of these cells by characterization of the TCRbeta repertoire. METHODS: TCRbeta repertoire was analyzed by reverse-transcription polymerase chain reaction/Southern blot and DNA sequencing. Clonality of T cells was examined in the lymphoid tissues and colons of TCRalpha(-/-) mice and interleukin 4-deficient TCRalpha(-/-) mice. In addition, an in vitro culture system using syngeneic colonic epithelial cells as antigens was used. RESULTS: The clonal expansion of a restricted subset of Vbeta8.2(+) T cells was characterized by conservation of a single negatively charged amino acid residue in the second position of the complementarity determining region 3 (CDR3). These T cells were observed in the diseased colon and appendix (cecal patch) of TCRalpha(-/-) mice, but not germfree TCRalpha(-/-) mice. Culture of polyclonal T cells from young TCRalpha(-/-) mice with colonic epithelial cells under T helper 2 conditions resulted in the survival of Vbeta8.2(+) T cells characterized by the same CDR3 pattern. In addition, the transfer of the cultivated T cells induced mild colitis in recombination activating gene 1 mutant mice. CONCLUSIONS: In the TCRalpha(-/-) mice, the development of colitis is associated with the presence of a restricted diversity of Vbeta8. 2(+) T-cell subsets characterized by a specific TCR motif. The limited diversity of lamina propria T cells that are derived from naive T cells expanded by reacting with luminal bacterial antigens is likely caused by the survival of these T cells after stimulation with self-antigens in the presence of a T helper 2 environment. PMID- 11040186 TI - Interleukin 15 mediates epithelial changes in celiac disease. AB - BACKGROUND & AIMS: Villous atrophy and crypt proliferation are key epithelial features of untreated celiac disease. We tried to define whether cytokines such as interleukin (IL)-15, IL-2, IL-4, and IL-7, which share chains of their receptors, could influence the epithelial modifications. METHODS: Duodenal biopsy specimens (14 treated and 13 untreated celiac patients, 7 controls) were cultured in vitro for 24 hours with or without gliadin (1 mg/mL), IL-15, IL-7, IL-4, or IL 2 (10 ng/mL). Tumor necrosis factor (TNF)-alpha and interferon (IFN)-gamma were also used in some specimens of untreated celiacs. Epithelial expression of Ki67, FAS, and transferrin receptor (TFR) was detected by immunohistochemistry, and apoptosis by TUNEL technique (percentage of positive enterocytes). IL-15-positive cells were detected by immunohistochemistry in celiac disease and control biopsy specimens; presence of IL-15 was also determined by semiquantitative polymerase chain reaction. RESULTS: Only IL-15 induced enterocyte expression of Ki67, TFR, and FAS in treated celiac (P<0.01 vs. medium) and enterocyte apoptosis in untreated celiac disease specimens. Anti-IL-15 monoclonal antibodies neutralized gliadin-induced enterocyte TFR and FAS expression in treated celiac and enterocyte apoptosis in untreated celiac disease specimens (P<0.05 vs. gliadin). IL-15-positive cells were increased in untreated celiacs (P<0.001 vs. treated celiacs and controls). CONCLUSIONS: IL-15 is involved in the modulation of epithelial changes in celiac disease, indicating that this cytokine has an unforeseen role in the pathologic manifestations of celiac disease. PMID- 11040187 TI - Leukotrienes induce cell-survival signaling in intestinal epithelial cells. AB - BACKGROUND & AIMS: Inflammatory bowel conditions, particularly ulcerative colitis, are associated with an increased incidence of neoplastic transformation. High levels of proinflammatory leukotrienes (LTs) and up-regulated expression of cyclooxygenase (COX)-2 are characteristic of inflammation. Moreover, COX-2 has been implicated in cell survival and early colon carcinogenesis. Other aspects of interest for intestinal cell viability are the levels of beta-catenin and the antiapoptotic protein Bcl-2. We investigated the possibility that LTs participate in the regulation of these survival factors. METHODS: We used the human intestinal epithelial cell line Int 407 and the rat intestinal epithelial cell line IEC-6. Immunoblotting was applied to ascertain protein expression and distribution, and enzyme immunoassay methodology was used to measure prostaglandin E(2) (PGE(2)) production. Apoptotic ability was assessed by trypan blue exclusion, Hoechst staining, DNA fragmentation, and a caspase-3 activity assay. RESULTS: LTD(4) and LTB(4), but not LTC(4), caused a time- and dose dependent increase in expression and/or membrane accumulation of COX-2, beta catenin, and Bcl-2, as well as PGE(2) production. Apoptosis assays showed that the effects of LTs on these transformation-associated proteins correlated well with the ability of these LTs to reduce programmed cell death. CONCLUSIONS: The results suggest that inflammatory conditions are associated with the expression and distribution of proteins that are characteristic of transformed cells; such conditions may involve a signaling mechanism comprising an altered rate of apoptosis. PMID- 11040188 TI - Environmental stress-induced gastrointestinal permeability is mediated by endogenous glucocorticoids in the rat. AB - BACKGROUND & AIMS: Abnormal presentation of luminal constituents to the mucosal immune system, caused by dysfunction of the intestinal epithelial barrier, is a candidate theory for the cause of Crohn's disease. Increased epithelial permeability is found in subgroups of patients at high risk for the development of Crohn's disease and has been found to precede disease recurrence. Clinical observations have suggested that disease recurrence can follow times of increased psychological stress, although the underlying mechanism remains obscure. We hypothesized that environmental stress increases gastrointestinal permeability. METHODS: We evaluated site-specific gastrointestinal permeability after application of graded levels of stress in rats. RESULTS: Increased epithelial permeability after stress was shown in all regions of the gastrointestinal tract and seemed to be mediated by adrenal corticosteroids. Stress-induced increases in epithelial permeability disappeared after adrenalectomy or pharmacologic blockade of glucocorticoid receptors. Dexamethasone treatment of control animals increased gastrointestinal permeability and mimicked the effects of stress. CONCLUSIONS: Psychological stress may increase gastrointestinal permeability, allowing luminal constituents access to the mucosal immune system. This provides a potential mechanism for the observation of stress-induced disease recurrence in Crohn's disease. PMID- 11040189 TI - Permeability of the rat small intestinal epithelium along the villus-crypt axis: effects of glucose transport. AB - BACKGROUND & AIMS: The aim of this study was to elucidate the permeability characteristics of the epithelium along the villus-crypt axis and investigate the effect of glucose transport on these characteristics along this axis. METHODS: The disappearance rates of (14)C-mannitol and (51)Cr-EDTA or (3)H-inulin were determined as clearance (Cl(x)) from a recirculating perfusion system of the jejunal lumen in anesthetized rats. Net fluid transport was varied over a large range by exchanging mannitol with glucose in the perfusate solution and by inhibition of nervously mediated secretory processes with hexamethonium. The perfusion rate was 0.5 or 0.2 mL/min. RESULTS: Cl(Man) enhanced significantly with increasing net fluid transport (secretion 8.50+/-1.88, to absorption 16.72+/ 1.75 microL x min(-1) x g(-1)) and with glucose perfusates. Cl(Cr-EDTA) was constant irrespective of net fluid transport and was reduced to insignificant values at a perfusion rate of 0.2 mL/min. Cl(In) was not different from zero. CONCLUSIONS: The absorbing apical part of the villus contains small pores (radius, <6 A) allowing passive transport via solvent drag of, e.g., monosaccharides, whereas the pores in the crypts are large (50-60 A) and inaccessible to the luminal content. The basal part of the villus contains medium sized pores (10-15 A) through which no solvent drag occurs. Active glucose transport in the rat mainly increases the number of small pores accessible for passive transport, whereas the size of these pores seems to stay constant. PMID- 11040190 TI - Neurokinin 1 and 2 receptors mediate cholera toxin secretion in rat jejunum. AB - BACKGROUND & AIMS: Substance P, a member of the tachykinin family, is a prosecretory neuropeptide distributed widely throughout the enteric nervous system. Implicated in inflammatory states, its role in enterotoxigenic water and electrolyte secretion is unclear. We assessed the effect of substance P antagonists and neurokinin receptor antagonists on cholera toxin-, Escherichia coli heat-labile enterotoxin (LT)-, and heat-stable enterotoxin (STa)-induced water secretion in an in vivo rat jejunal perfusion model. METHODS: Anesthetized adult male Wistar rats were pretreated with substance P antagonists (D-Pro(2), D Trp(79), substance P, 0.1-3.0 mg/kg; or CP 96,345/4, 0.3-3 mg/kg) or neurokinin (NK)-1 (sendide, 1.0 mg/kg), NK-2 (GR83074, 1.0 mg/kg), or NK-3 ([Trp(7),betaAla(8)]NKA(4-10), 1.0 mg/kg) receptor antagonists. In a subgroup, extrinsic sensory afferents were ablated by pretreatment with capsaicin. Jejunal perfusion, with a plasma electrolyte solution containing a nonabsorbable marker, was undertaken after exposure to cholera toxin (25 microg), LT (25 microg), STa (200 microg/L), or saline. RESULTS: Cholera toxin-induced water and electrolyte secretion was inhibited by the substance P antagonists and the NK-1 and NK-2 receptor antagonists, but not by the NK-3 receptor antagonist or by pretreatment with capsaicin. Neither LT- nor STa-induced secretions were affected by the pretreatments. CONCLUSIONS: Prosecretory pathways involving NK-1 and NK-2 receptors specifically mediate the actions of cholera toxin in the small intestine. PMID- 11040191 TI - E-cadherin and adenomatous polyposis coli mutations are synergistic in intestinal tumor initiation in mice. AB - BACKGROUND & AIMS: Inactivation of the adenomatous polyposis coli (APC) gene is observed at early stages of intestinal tumor formation, whereas loss of E cadherin is usually associated with tumor progression. Because both proteins compete for the binding to beta-catenin, an essential component of the Wnt signaling pathway, reduction of E-cadherin levels in an Apc mouse model could influence both tumor initiation and progression. In addition, loss or haploinsufficiency of E-cadherin may affect tumorigenesis by altering its cell adhesive and associated functions. METHODS: Apc1638N mice were bred with animals carrying a targeted E-cadherin knockout mutation. RESULTS: Double heterozygous animals showed a significant 9-fold and 5-fold increase of intestinal and gastric tumor numbers, respectively, compared with Apc1638N animals. The intestinal tumors of both groups showed no significant differences in grading and staging. Loss of heterozygosity analysis at the Apc and E-cadherin loci in both intestinal and gastric Apc(+/1638N)/E-cad(+/-) tumors revealed loss of the wild-type Apc allele in most cases, whereas the wild-type E-cadherin allele was always retained. This was supported by a positive, although reduced, staining for E cadherin of intestinal tumor sections. CONCLUSIONS: Introduction of the E cadherin mutation in Apc1638N animals enhances Apc-driven tumor initiation without clearly affecting tumor progression. PMID- 11040192 TI - Orphanin FQ causes contractions via inhibiting purinergic pathway in the rat colon. AB - BACKGROUND & AIMS: We have previously shown that orphanin FQ (OFQ) preferentially stimulates muscle contraction in the rat colon. However, the mechanism of action of OFQ remains unclear. METHODS: We studied the effects of OFQ on muscle contractions and inhibitory junction potentials (IJPs) in rat colon. The site of action of OFQ was also investigated by in situ hybridization of OFQ receptors. RESULTS: OFQ (10(-10) to 10(-6) mol/L) caused circular muscle contractions that were blocked by tetrodotoxin (10(-7) mol/L), suggesting the contractions were nerve mediated. Suramin (a nonselective P(2)-purinoceptor antagonist; 10(-4) mol/L) and reactive blue 2 (a P(2Y)-purinoceptor antagonist; 3 x 10(-5) mol/L), but not pyridoxalphosphate-6-azophenyl-2',4' disulfonic acid (PPADS; a P(2X) purinoceptor antagonist; 3 x 10(-5) mol/L), abolished OFQ-induced colonic contractions. Focal stimulation of interganglionic fiber tracts evoked biphasic IJPs in colonic circular muscle cells. Suramin and reactive blue 2 inhibited the peak amplitude of the IJP, whereas PPADS had no effect. Cumulative addition of OFQ (10(-10) to 10(-6 )mol/L) significantly inhibited the IJPs. In situ hybridization revealed that OFQ receptor messenger RNA was expressed in the colonic myenteric plexus but not in the smooth muscle cells, suggesting that the site of action of OFQ is neuronal. CONCLUSIONS: These results suggest that OFQ causes muscle contractions by inhibiting purinergic inhibitory motorneurons in the rat colon. PMID- 11040193 TI - Hepatitis virus infections in heart transplant recipients: epidemiology, natural history, characteristics, and impact on survival. AB - BACKGROUND & AIMS: We have observed a high prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection in heart transplant recipients (HTRs). The aim of this study was to assess the epidemiology, natural history, and clinical and biological characteristics of viral hepatitis in HTRs. METHODS: From 1983 to 1992, 874 patients underwent heart transplantation at the Pitie-Salpetriere Hospital, Paris, France, 459 of whom qualified for analysis. A total of 140 patients had posttransplantation hepatitis B, C, or non-A-E. Sixty-nine patients developed HBV infection, 49 HCV infection, 11 HBV-HCV coinfection, and 11 non-A-E hepatitis. RESULTS: HBV was transmitted nosocomially from patient to patient, most likely during endomyocardial biopsies. HCV was mainly transmitted through blood transfusions or the transplanted organ. Clinical and biological findings after 2 years of follow-up showed that 3 patients with an HBV genotype A precore mutant had severe or subfulminant hepatitis and that patients with HBV and HCV infection always progressed to chronicity. In general, patients had mild alanine aminotransferase level increases, a high level of viral replication, and few severe histologic lesions, except for patients infected by precore HBV mutants. Patients coinfected by HBV and HCV tended to have more severe liver lesions. The survival rate 5 years after transplantation in patients with viral hepatitis (HBV, 81%; HCV, 89%; HBV and HCV coinfection, 100%; non-A-E hepatitis, 73%) was similar to that in patients without liver test abnormalities (76%). The actuarial survival curve was also similar in patients with or without liver test abnormalities. CONCLUSIONS: In our experience, histologic liver lesions do not progress rapidly in patients with post-heart transplant infection caused by HBV or HCV. HBV or HCV infection seems to have little impact on the 5-year survival rate of HTRs. PMID- 11040194 TI - Differential HFE allele expression in hemochromatosis heterozygotes. AB - BACKGROUND & AIMS: Hereditary hemochromatosis is associated with C282Y homozygosity. Some heterozygotes may also present with abnormal iron parameters. However, the precise role of H63D and C282Y mutations in iron overload is poorly understood. We investigated the level of expression of the mutated and unmutated HFE alleles in these heterozygous patients. METHODS: We studied the expression of HFE messenger RNAs in peripheral blood mononuclear cells from 34 heterozygotes using reverse-transcription polymerase chain reaction (PCR) followed by enzymatic digestion or sequence analysis of the PCR products, which allows relative quantification of mutated and unmutated transcripts. HFE proteins were quantified by Western blotting in Epstein-Barr virus-immortalized lymphocyte extracts from 2 C282Y and H63D homozygotes and a compound heterozygote. RESULTS: (187C > G; H63D) mutated transcripts predominated in H63D and compound heterozygotes and the normal transcripts in C282Y heterozygotes. The amount of HFE protein was increased in the H63D homozygotes and the compound heterozygote compared with the C282Y homozygotes. In addition, we found a new mutation at codon 282 (C282S) associated with severe iron overload. CONCLUSIONS: We demonstrate the existence of differential allelic expression of the HFE alleles, suggesting that the (187C > G; H63D) mutation plays a role in the disease expression in H63D heterozygotes, in particular when associated with environmental or host factors. PMID- 11040195 TI - Pancreatic involvement in von Hippel-Lindau disease. The Groupe Francophone d'Etude de la Maladie de von Hippel-Lindau. AB - BACKGROUND & AIMS: Pancreatic involvement in von Hippel-Lindau (VHL) disease, a genetic disorder with a dominant mode of inheritance affecting various organs, has rarely been studied. We assessed the prevalence, type of lesions, natural history, and impact of pancreatic involvement in patients with VHL. METHODS: A total of 158 consecutive patients from 94 families with VHL disease were studied in a prospective French collaborative study. All patients underwent systematic screening for VHL lesions, including computerized tomography (CT) scanning of the pancreas reviewed by an experienced radiologist. Clinical data, investigations, and treatments performed were also reviewed. RESULTS: Pancreatic involvement was observed in 122 patients (77.2%) and included true cysts (91.1%), serous cystadenomas (12.3%), neuroendocrine tumors (12.3%), or combined lesions (11.5%). The pancreas was the only organ affected in 7.6% of patients. Patients with pancreatic lesions had fewer pheochromocytomas than those without (14/122 vs. 16/36; P<0.0001), and patients with neuroendocrine pancreatic tumors had renal involvement less often than those without (8/99 vs. 6/20; P = 0.013). None of the patients with neuroendocrine tumors had symptoms of hormonal hypersecretion. Pancreatic lesions evolved in half of patients but required specific treatment in only 10 (8.2%) when they were symptomatic or for the resection of large neuroendocrine tumors. CONCLUSIONS: Pancreatic involvement is seen in most patients with VHL disease. Although symptoms are rare, specific treatment of pancreatic lesions is required in selected patients, mainly those with neuroendocrine tumors. PMID- 11040196 TI - Circulating soluble vascular adhesion protein 1 accounts for the increased serum monoamine oxidase activity in chronic liver disease. AB - BACKGROUND & AIMS: Vascular adhesion protein 1 (VAP-1) is an endothelial glycoprotein that supports adhesion of lymphocytes to hepatic endothelium and has sequence homology with semicarbazide-sensitive amine oxidases (SSAOs). We investigated whether soluble VAP-1 (sVAP-1) displays SSAO activity and thereby accounts for increased monoamine oxidase activity in the serum of patients with liver diseases. METHODS: sVAP-1 concentration and SSAO activity were measured in peripheral, hepatic, and portal blood and in bile from patients with liver disease and in peripheral blood of control subjects, using enzyme-linked immunosorbent assay and enzymatic assays. RESULTS: sVAP-1 concentration (mean [+/ SE], 143. 67 [34.97-92.67] ng/mL) and SSAO activity (18.8 [12.0-24.6] nmol. mL( 1). h(-1)) were significantly increased in chronic liver diseases compared with healthy controls (87.1 [53.5-127] ng/mL [P<0.001] and 10.7 [6.5-12.7] nmol. mL( 1) x h(-1) [P<0.05]) but not in massive necrosis caused by paracetamol poisoning (109 [80.3-140] ng/mL and 8.9 [5.7-12.3] nmol. mL(-1) x h(-1)). sVAP-1 correlated with serum transaminase and bilirubin but not with creatinine. In 5 paired samples, sVAP-1 concentration was higher in hepatic (median, 113 [range, 53-122]) than in portal vein (102 [42-109]; 2P<0.05), and was not detected in bile. There was a highly significant correlation between serum sVAP-1 and SSAO activity in normal subjects, patients with acute liver failure, and those with chronic liver disease (r = 0.895; P<0.001). When serum was depleted of sVAP-1 by immunoaffinity chromatography, SSAO activity was eliminated. CONCLUSIONS: sVAP-1 levels are increased in chronic liver disease, and sVAP-1 is likely derived from the liver. Serum sVAP-1 displays SSAO activity and accounts for most of the monoamine oxidase activity in human serum. PMID- 11040197 TI - Mouse alpha-fetoprotein-specific DNA-based immunotherapy of hepatocellular carcinoma leads to tumor regression in mice. AB - BACKGROUND & AIMS: alpha-Fetoprotein (AFP) is a tumor-associated protein that is frequently expressed at high levels in hepatocellular carcinoma (HCC). The aim of the study was to characterize self-reactive cytotoxic T lymphocytes (CTLs) directed against murine AFP (mAFP) after DNA-based immunization in mice. METHODS: To study CTL responses, mAFP-expressing recombinant vaccinia viruses were generated. An HCC tumor model was established in C57L/J mice by injection of syngeneic endogenously mAFP-expressing Hepa1-6 cells. RESULTS: Gene gun and intramuscular coimmunizations of DNA expression vectors encoding mAFP with plasmids encoding murine interleukin (IL)-12, granulocyte-macrophage colony stimulating factor, or IL-18 induced weak CTL activity against mAFP in different mouse strains. Some mice developed anti-mAFP antibody responses, suggesting breaking of immunologic ignorance. No hepatocyte damage was detectable despite low-level endogenous hepatic mAFP expression. Therapeutic immunizations of mice bearing mAFP-expressing murine HCCs induced partial regression of tumors. A significant survival benefit was observed in mice immunized with mAFP expression vector DNA but not in untreated mice or in mice immunized with mock/cytokine plasmid DNA. CONCLUSIONS: The data show that AFP may be used as a potential self tumor antigen to induce CTL and CD4(+) T cell-mediated regression of AFP expressing HCC by DNA-based immunization. PMID- 11040198 TI - Molecular identification and functional characterization of Mdr1a in rat cholangiocytes. AB - BACKGROUND & AIMS: The multidrug resistance P-glycoprotein 170 gene products (mdr1a and 1b) are glycosylated plasma membrane proteins that function as adenosine triphosphate-dependent transmembrane export pumps for lipophilic xenobiotics of widely different structure. We assessed whether these P glycoproteins are functionally expressed in cholangiocytes. METHODS: A reverse transcription polymerase chain reaction was performed on RNA from a normal rat cholangiocyte cell line using mdr1-specific primers. Northern and Western blot analyses were performed on cholangiocytes immunoisolated from 2-week bile duct ligated rats and cholangiocytes and isolated cholangiocyte membrane subfractions, respectively. Functional assays were performed in isolated bile duct units from bile duct-ligated rats and incubated with rhodamine 123, a P-glycoprotein substrate, with or without the P-glycoprotein inhibitors verapamil or GF120918. RESULTS: A 400-base pair fragment with 99% homology to the cytosolic domain of rat intestinal mdr1a (5' 1953-2350 3') was identified that hybridized to a 5.2 kilobase RNA transcript in a normal rat cholangiocyte cell line, isolated rat cholangiocytes, and ileum. Western analysis localized mdr1 to the apical membrane of cholangiocytes. Confocal microscopy showed active secretion of rhodamine 123 into the lumen of isolated bile duct units that was abolished by vanadate and P glycoprotein competitive antagonists, verapamil and GF120918, in a dose-dependent manner. CONCLUSIONS: These findings provide the first molecular and functional evidence for the expression of mdr1a on the luminal membrane of cholangiocytes, where it may have a protective role. PMID- 11040199 TI - Kinesin is involved in regulation of rat pancreatic amylase secretion. AB - BACKGROUND & AIMS: Kinesin has recently been localized to zymogen granules of pancreatic acini and is suggested to participate in exocytosis of exocrine pancreas. We examined the function of kinesin in regulated exocytosis of pancreatic acini in this study. METHODS: Kinesin function in exocytosis was examined by introducing hexahistidine-tagged recombinant kinesin protein and antikinesin monoclonal antibody into streptolysin-O-permeabilized acini. Intracellular localization of introduced recombinant kinesin was investigated by immunohistochemistry. Interaction between recombinant kinesin and the microtubule network was confirmed by nocodazole pretreatment of acini. Kinesin regulation by secretagogues was investigated by examining their effect on adenosine triphosphatase (ATPase) activity of endogenous kinesin. RESULTS: Recombinant kinesin enhanced calcium-stimulated amylase release from streptolysin-O permeabilized acini. Introduced recombinant kinesin was localized to both the microtubule network and zymogen granule. Nocodazole pretreatment of acini abolished the enhancing effect of recombinant kinesin on calcium-stimulated amylase release. Antikinesin antibody inhibited amylase release stimulated by the combination of calcium and cyclic adenosine monophosphate (cAMP) but not that stimulated by calcium alone. Secretin and 8-bromo-cAMP increased ATPase activity of endogenous kinesin. CONCLUSIONS: Kinesin plays a stimulatory role in regulated exocytosis of pancreatic acini and is involved in stimulus-secretion coupling through a cAMP-dependent pathway. PMID- 11040200 TI - Treatment of fistulizing Crohn's disease. AB - The appropriate treatment of patients with fistulas in the setting of Crohn's disease requires a knowledge of the specific medical and surgical literature of fistulizing Crohn's. The patient with symptomatic fistulizing Crohn's disease may respond differently to specific medical therapy than a patient with symptomatic obstructing Crohn's disease. Certain medications that are useful for the treatment of patients with obstructive Crohn's disease may not be helpful in the treatment of fistulas in patients with fistulizing Crohn's disease (e.g., corticosteroids and mesalamine); in fact, some medications are believed to be detrimental (e.g., corticosteroids). Few studies have been performed to assess the efficacy of specific medications on fistulas directly. To date, there has been only one published prospective randomized controlled trial that was designed to assess the efficacy and safety of a specific medication on fistulas in patients with Crohn's disease; it showed clinical efficacy over placebo in a statistically significant manner. The judicious use of surgery remains an integral part of the management of certain presentations of fistulizing Crohn's disease, and the appropriate integration of surgical and medical therapy is of paramount importance in the management of these patients. This review provides an overview of pertinent medical and surgical literature as it pertains to management of patients with fistulizing Crohn's disease. PMID- 11040201 TI - Tumor necrosis factor: biology and therapeutic inhibitors. PMID- 11040202 TI - 6-Mercaptopurine in maintaining remission in Crohn's disease: An old friend becomes a new hero. PMID- 11040203 TI - Touch and go: mediating cell-to-cell interactions and Wnt signaling in gastrointestinal tumor formation. PMID- 11040204 TI - Barrett's esophagus: the long and the short of it. PMID- 11040205 TI - Turning on T-cell death and turning off Crohn's disease. PMID- 11040206 TI - The benefits of dietary fiber on glycemic control in type 2 diabetes: relevance to gastroenterologists. PMID- 11040208 TI - Surgery of the anus, rectum and colon PMID- 11040207 TI - Beta-blocker plus nitrate for primary prophylaxis of variceal bleeding. PMID- 11040209 TI - Orchestrated response: a symphony of transcription factors for gene control. PMID- 11040210 TI - The function of Xenopus Bloom's syndrome protein homolog (xBLM) in DNA replication. AB - The Bloom's syndrome gene (BLM) plays a pivotal role in the maintenance of genomic stability in somatic cells. It encodes a DNA helicase (BLM) of the RecQ family, but the exact function of BLM remains elusive. To study this question, we have cloned the BLM homolog of the frog Xenopus laevis (xBLM) and have raised antibodies to it. Immunodepletion of xBLM from a Xenopus egg extract severely inhibits the replication of DNA in reconstituted nuclei. Moreover, the inhibition can be rescued by the addition of the recombinant xBLM protein. These results provide the first direct evidence that BLM plays an important role in DNA replication, suggesting that Bloom's syndrome may be the consequence of defective DNA replication. PMID- 11040211 TI - Defective neurogenesis resulting from DNA ligase IV deficiency requires Atm. AB - Ataxia telangiectasia results from mutations of ATM and is characterized by severe neurodegeneration and defective responses to DNA damage. Inactivation of certain DNA repair genes such as DNA ligase IV results in massive neuronal apoptosis and embryonic lethality in the mouse, indicating the occurrence of endogenously formed DNA double-strand breaks during nervous system development. Here we report that Atm is required for apoptosis in all areas of the DNA ligase IV-deficient developing nervous system. However, Atm deficiency failed to rescue deficits in immune differentiation in DNA ligase IV-null mice. These data indicate that ATM responds to endogenous DNA lesions and functions during development to eliminate neural cells that have incurred genomic damage. Therefore, ATM could be important for preventing accumulation of DNA-damaged cells in the nervous system that might eventually lead to the neurodegeneration observed in ataxia telangiectasia. PMID- 11040212 TI - p21 is a transcriptional target of HOXA10 in differentiating myelomonocytic cells. AB - The myeolomonocytic cell line U937 differentiates into macrophages in response to a variety of agents. Several genes including the cyclin-dependent kinase inhibitor p21(waf1/cip1) and the homeobox gene transcription factor HOXA10 are induced at the onset of differentiation. Ectopic expression of either gene results in U937 differentiation. In this paper, we describe a mechanism by which p21 and HOXA10 may act in concert, where HOXA10 can bind directly to the p21 promoter and, together with its trimeric partners PBX1 and MEIS1, activate p21 transcription, resulting in cell cycle arrest and differentiation. These experiments for the first time identify p21 as a selective target for a HOX protein and link the differentiative properties of a transcription factor and a cell cycle inhibitor. PMID- 11040213 TI - Dmrt1, a gene related to worm and fly sexual regulators, is required for mammalian testis differentiation. AB - The only molecular similarity in sex determination found so far among phyla is between the Drosophila doublesex (dsx) and Caenorhabditis elegans mab-3 genes. dsx and mab-3 contain a zinc finger-like DNA-binding motif called the DM domain, perform several related regulatory functions, and are at least partially interchangeable in vivo. A DM domain gene called Dmrt1 has been implicated in male gonad development in a variety of vertebrates, on the basis of embryonic expression and chromosomal location. Such evidence is highly suggestive of a conserved role(s) for Dmrt1 in vertebrate sexual development, but there has been no functional analysis of this gene in any species. Here we show that murine Dmrt1 is essential for postnatal testis differentiation, with mutant phenotypes similar to those caused by human chromosome 9p deletions that remove the gene. As in the case of 9p deletions, Dmrt1 is dispensable for ovary development in the mouse. Thus, as in invertebrates, a DM domain gene regulates vertebrate male development. PMID- 11040214 TI - CPEB proteins control two key steps in spermatogenesis in C. elegans. AB - Cytoplasmic polyadenylation element binding (CPEB) proteins bind to and regulate the translation of specific mRNAs. CPEBs from Xenopus, Drosophila, and Spisula participate in oogenesis. In this report, we examine the biological roles of all identifiable CPEB homologs in a single organism, Caenorhabditis elegans. We find four homologs in the C. elegans genome: cbp-1, cpb-2, cpb-3, and fog-1. Surprisingly, two homologs, CPB-1 and FOG-1, have key functions in spermatogenesis and are dispensable for oogenesis. CPB-2 and CPB-3 also appear not to be required for oogenesis. CPB-1 is essential for progression through meiosis: cpb-1(RNAi) spermatocytes fail to undergo the meiotic cell divisions. CPB-1 protein is present in the germ line just prior to overt spermatogenesis; once sperm differentiation begins, CPB-1 disappears. CPB-1 physically interacts with FBF, another RNA-binding protein and 3' UTR regulator. In addition to its role in controlling the sperm/oocyte switch, we find that FBF also appears to be required for spermatogenesis, consistent with its interaction with CPEB. A second CPEB homolog, FOG-1, is required for specification of the sperm fate. The fog-1 gene produces fog-1(L) and fog-1(S) transcripts. The fog-1(L) RNA is enriched in animals making sperm and is predicted to encode a larger protein; fog-1(S) RNA is enriched in animals making oocytes and is predicted to encode a smaller protein. The relative abundance of the two mRNAs is controlled temporally during germ-line development and by the sex determination pathway in a fashion that suggests that the fog-1(L) species encodes the active form. In sum, our results demonstrate that, in C. elegans, two CPEB proteins have distinct functions in the germ line, both in spermatogenesis: FOG-1 specifies the sperm cell fate and CPB-1 executes that decision. PMID- 11040215 TI - Raf induces TGFbeta production while blocking its apoptotic but not invasive responses: a mechanism leading to increased malignancy in epithelial cells. AB - c-Raf-1 is a major effector of Ras proteins, responsible for activation of the ERK MAP kinase pathway and a critical regulator of both normal growth and oncogenic transformation. Using an inducible form of Raf in MDCK cells, we have shown that sustained activation of Raf alone is able to induce the transition from an epithelial to a mesenchymal phenotype. Raf promoted invasive growth in collagen gels, a characteristic of malignant cells; this was dependent on the operation of an autocrine loop involving TGFbeta, whose secretion was induced by Raf. TGFbeta induced growth inhibition and apoptosis in normal MDCK cells: Activation of Raf led to inhibition of the ability of TGFbeta to induce apoptosis but not growth retardation. ERK has been reported previously to inhibit TGFbeta signaling via phosphorylation of the linker region of Smads, which prevents their translocation to the nucleus. However, we found no evidence in this system that ERK can significantly influence the function of Smad2, Smad3, and Smad4 at the level of nuclear translocation, DNA binding, or transcriptional activation. Instead, strong activation of Raf caused a broad protection of these cells from various apoptotic stimuli, allowing them to respond to TGFbeta with increased invasiveness while avoiding cell death. The Raf-MAP kinase pathway thus synergizes with TGFbeta in promoting malignancy but does not directly impair TGFbeta-induced Smad signaling. PMID- 11040217 TI - High-resolution localization of Drosophila Spt5 and Spt6 at heat shock genes in vivo: roles in promoter proximal pausing and transcription elongation. AB - Recent studies have demonstrated roles for Spt4, Spt5, and Spt6 in the regulation of transcriptional elongation in both yeast and humans. Here, we show that Drosophila Spt5 and Spt6 colocalize at a large number of transcriptionally active chromosomal sites on polytene chromosomes and are rapidly recruited to endogenous and transgenic heat shock loci upon heat shock. Costaining with antibodies to Spt6 and to either the largest subunit of RNA polymerase II or cyclin T, a subunit of the elongation factor P-TEFb, reveals that all three factors have a similar distribution at sites of active transcription. Crosslinking and immunoprecipitation experiments show that Spt5 is present at uninduced heat shock gene promoters, and that upon heat shock, Spt5 and Spt6 associate with the 5' and 3' ends of heat shock genes. Spt6 is recruited within 2 minutes of a heat shock, similar to heat shock factor (HSF); moreover, this recruitment is dependent on HSF. These findings provide support for the roles of Spt5 in promoter-associated pausing and of Spt5 and Spt6 in transcriptional elongation in vivo. PMID- 11040216 TI - Spt5 and spt6 are associated with active transcription and have characteristics of general elongation factors in D. melanogaster. AB - The Spt4, Spt5, and Spt6 proteins are conserved throughout eukaryotes and are believed to play critical and related roles in transcription. They have a positive role in transcription elongation in Saccharomyces cerevisiae and in the activation of transcription by the HIV Tat protein in human cells. In contrast, a complex of Spt4 and Spt5 is required in vitro for the inhibition of RNA polymerase II (Pol II) elongation by the drug DRB, suggesting also a negative role in vivo. To learn more about the function of the Spt4/Spt5 complex and Spt6 in vivo, we have identified Drosophila homologs of Spt5 and Spt6 and characterized their localization on Drosophila polytene chromosomes. We find that Spt5 and Spt6 localize extensively with the phosphorylated, actively elongating form of Pol II, to transcriptionally active sites during salivary gland development and upon heat shock. Furthermore, Spt5 and Spt6 do not colocalize widely with the unphosphorylated, nonelongating form of Pol II. These results strongly suggest that Spt5 and Spt6 play closely related roles associated with active transcription in vivo. PMID- 11040218 TI - Different human TFIIIB activities direct RNA polymerase III transcription from TATA-containing and TATA-less promoters. AB - Transcription initiation at RNA polymerase III promoters requires transcription factor IIIB (TFIIIB), an activity that binds to RNA polymerase III promoters, generally through protein-protein contacts with DNA binding factors, and directly recruits RNA polymerase III. Saccharomyces cerevisiae TFIIIB is a complex of three subunits, TBP, the TFIIB-related factor BRF, and the more loosely associated polypeptide beta("). Although human homologs for two of the TFIIIB subunits, the TATA box-binding protein TBP and the TFIIB-related factor BRF, have been characterized, a human homolog of yeast B(") has not been described. Moreover, human BRF, unlike yeast BRF, is not universally required for RNA polymerase III transcription. In particular, it is not involved in transcription from the small nuclear RNA (snRNA)-type, TATA-containing, RNA polymerase III promoters. Here, we characterize two novel activities, a human homolog of yeast B("), which is required for transcription of both TATA-less and snRNA-type RNA polymerase III promoters, and a factor equally related to human BRF and TFIIB, designated BRFU, which is specifically required for transcription of snRNA-type RNA polymerase III promoters. Together, these results contribute to the definition of the basal RNA polymerase III transcription machinery and show that two types of TFIIIB activities, with specificities for different classes of RNA polymerase III promoters, have evolved in human cells. PMID- 11040219 TI - The alpha subunit of E. coli RNA polymerase activates RNA binding by NusA. AB - The Escherichia coli NusA protein modulates pausing, termination, and antitermination by associating with the transcribing RNA polymerase core enzyme. NusA can be covalently cross-linked to nascent RNA within a transcription complex, but does not bind RNA on its own. We have found that deletion of the 79 carboxy-terminal amino acids of the 495-amino-acid NusA protein allows NusA to bind RNA in gel mobility shift assays. The carboxy-terminal domain (CTD) of the alpha subunit of RNA polymerase, as well as the bacteriophage lambda N gene antiterminator protein, bind to carboxy-terminal regions of NusA and enable full length NusA to bind RNA. Binding of NusA to RNA in the presence of alpha or N involves an amino-terminal S1 homology region that is otherwise inactive in full length NusA. The interaction of the alpha-CTD with full-length NusA stimulates termination. N may prevent termination by inducing NusA to interact with N utilization (nut) site RNA rather than RNA near the 3' end of the nascent transcript. Sequence analysis showed that the alpha-CTD contains a modified helix hairpin-helix motif (HhH), which is also conserved in the carboxy-terminal regions of some eubacterial NusA proteins. These HhH motifs may mediate protein protein interactions in NusA and the alpha-CTD. PMID- 11040220 TI - On the sequencing of the human genome. PMID- 11040221 TI - Possible locus on chromosome 18q influencing postural systolic blood pressure changes. AB - We conducted a genome-wide scan for quantitative trait loci influencing the systolic blood pressure, diastolic blood pressure, and pulse responses to a postural challenge in 498 white sibling-pairs from the Hypertension Genetic Epidemiology Network, a multicenter study of the genetic susceptibility to hypertension. All participants were hypertensive (systolic blood pressure >/=140 mm Hg, diastolic blood pressure >/=90 mm Hg, or on antihypertensive medications) with diagnosis before age 60. Blood pressure and pulse were measured by an oscillometric method after a 5-minute rest in a supine position and again immediately on standing. The genome scan included a total of 387 autosomal short tandem-repeat polymorphisms typed by the National Heart, Lung, and Blood Institute Mammalian Genotyping Service at Marshfield. We used multipoint variance components linkage analysis to identify possible quantitative trait loci influencing postural change phenotypes after adjusting for sex, age, and use of antihypertensive medications. There was suggestive evidence for linkage on chromosome 18q for the postural systolic blood pressure response (maximum logarithm of the odds score=2.6 at 80 centiMorgans). We also observed a maximum logarithm of the odds score of 1.9 for the systolic blood pressure response and 1.7 for the diastolic blood pressure response on chromosome 6p. The marker that demonstrated the strongest evidence for linkage for the systolic blood pressure response (D18S858) lies within 20 centiMorgans of a marker previously linked to rare familial orthostatic hypotensive syndrome. Our findings indicate that there may be 1 or more genes on chromosome 18q that regulate systolic blood pressure during the physiological recovery period after a postural stressor. PMID- 11040223 TI - Arterial stiffness as underlying mechanism of disagreement between an oscillometric blood pressure monitor and a sphygmomanometer. AB - Oscillometric blood pressure devices tend to overestimate systolic blood pressure and underestimate diastolic blood pressure compared with sphygmomanometers. Recent studies indicate that discrepancies in performance between these devices may differ between healthy and diabetic subjects. Arterial stiffness in diabetics could be the underlying factor explaining these differences. We studied differences between a Dinamap oscillometric blood pressure monitor and a random zero sphygmomanometer in relation to arterial stiffness in 1808 healthy elderly subjects. The study was conducted within the Rotterdam Study, a population-based cohort study of subjects aged 55 years and older. Systolic and diastolic blood pressure differences between a Dinamap and a random-zero sphygmomanometer were related to arterial stiffness, as measured by carotid-femoral pulse wave velocity. Increased arterial stiffness was associated with higher systolic and diastolic blood pressure readings by the Dinamap compared with the random-zero sphygmomanometer, independent of age, gender, and average mean blood pressure level of both devices. The beta-coefficient (95% CI) was 0.25 (0.00 to 0.50) mm Hg/(m/s) for the systolic blood pressure difference and 0.35 (0.20 to 0.50) mm Hg/(m/s) for the diastolic blood pressure difference. The results indicate that a Dinamap oscillometric blood pressure device, in comparison to a random-zero sphygmomanometer, overestimates systolic and diastolic blood pressure readings in subjects with stiff arteries. PMID- 11040222 TI - Evidence for a gene influencing blood pressure on chromosome 17. Genome scan linkage results for longitudinal blood pressure phenotypes in subjects from the framingham heart study. AB - Hypertension is a leading cause of morbidity and mortality. Efforts to identify hypertension genes have focused on 3 approaches: mendelian disorders, candidate genes, and genome-wide scans. Thus far, these efforts have not identified genes that contribute substantively to overall blood pressure (BP) variation in the community. A 10-centiMorgan (cM) density genome-wide scan was performed in the largest families from 2 generations of Framingham Heart Study participants. Heritability and linkage for long-term mean systolic and diastolic BP phenotypes were analyzed by use of SOLAR software. Heritability estimates were based on BP measurements in 1593 families. Genotyping was performed on 1702 subjects from 332 large families, and BP data were available for 1585 (93%) genotyped subjects who contributed 12 588 longitudinal BP observations. The mean age was 47 years, and mean BP was 127/80 (systolic/diastolic) mm Hg. Long-term systolic and diastolic BP phenotypes had high heritability estimates, 0.57 and 0.56, respectively. For systolic BP, multipoint log-of-the-odds (LOD) scores >2.0 were located on chromosome 17 at 67 cM (LOD 4.7, P=0.0000016) and 94 cM (LOD 2.2). For diastolic BP, LOD scores >2.0 were identified on chromosome 17 (74 cM, LOD 2.1) and chromosome 18 (7 cM, LOD 2.1). Using a genome-wide scan, we found strong evidence for a BP quantitative trait locus on chromosome 17. Follow-up studies are warranted to identify the gene or genes in this quantitative trait locus that influence BP. Such knowledge could extend our understanding of the genetic basis of essential hypertension and have implications for the evaluation and treatment of patients with high BP. PMID- 11040224 TI - Impact of arterial stiffening on left ventricular structure. AB - Aging of the vasculature results in arterial stiffening and an increase in systolic and pulse pressures. Although pressure load is a stimulus for left ventricular hypertrophy, the extent to which vascular stiffening per se, independent of blood pressure, influences left ventricular structure is uncertain. Two hundred seventy-six subjects (79 normotensive and 197 otherwise healthy hypertensive individuals) underwent echocardiography to assess left ventricular structure. Arterial stiffness was estimated by the pressure independent stiffness index, beta, and the pressure-dependent elastic modulus derived from simultaneous carotid ultrasound and applanation tonometry. Systemic arterial compliance (the inverse of stiffness) was estimated by the arterial compliance index. In multivariate analysis, beta was related to age (P<0.001) and smoking history (P<0.01) but not mean pressure, whereas elastic modulus was related to age and mean pressure (both P<0.001). The arterial compliance index was only related to age. Whereas systolic and diastolic pressures and the elastic modulus were positively associated with left ventricular mass (all P<0.001), primarily because of increases in wall thicknesses, beta and the arterial compliance index bore no relation to left ventricular mass. beta was inversely related to chamber diameter and directly related to left ventricular relative wall thickness, the ratio of wall thickness to chamber radius. Younger and older hypertensive subjects had comparable left ventricular mass, despite higher systolic and pulse pressures in the older group, whereas older hypertensives had higher mean relative wall thickness, associated with a significant increase in arterial stiffness (beta, 7.06 versus 5.17; elastic modulus, 595 versus 437 dyne/cm(2) x10(-6)) and reduction in the arterial compliance index (0.87 versus 1.05 mL/mm Hg per square meter) (all P<0.001). Thus, the extent to which arterial stiffness relates to left ventricular hypertrophy is dependent on the method by which arterial stiffness is estimated. Pressure-dependent methods show an association with left ventricular hypertrophy, whereas the pressure-independent stiffness index, beta, and the arterial compliance index are most strongly associated with aging and left ventricular concentric remodeling but not hypertrophy. PMID- 11040225 TI - Cardiac aldosterone production in genetically hypertensive rats. AB - Aldosterone is synthesized in extra-adrenal tissues, both blood vessels and brain. We undertook the present study to determine whether the rat heart produces aldosterone and to investigate the effects of adrenalectomy, ACE inhibition, and angiotensin II on aldosterone synthesis in the heart. To clarify the pathophysiological role of cardiac aldosterone in the hypertensive heart, we compared the synthesis of aldosterone in the hearts of stroke-prone spontaneously hypertensive rats (SHRSP) with that in Wistar-Kyoto rats. The effects of the aldosterone antagonist spironolactone on myocardial hypertrophy in adrenalectomized SHRSP were also studied. Isolated rat hearts were perfused for 2 hours, and the perfusate was analyzed with HPLC and mass spectrometry. The activity of aldosterone synthase was estimated on the basis of the conversion of [(14)C]deoxycorticosterone to [(14)C]aldosterone. The levels of aldosterone synthase gene (CYP11B2) mRNA were determined with competitive polymerase chain reaction. Aldosterone production, the activity of aldosterone synthase, and the expression of CYP11B2 mRNA were increased in hearts from adrenalectomized rats and rats treated with angiotensin II. ACE inhibitors decreased cardiac aldosterone synthesis. Cardiac aldosterone, aldosterone synthase activity, and CYP11B2 mRNA levels in hearts from 2- and 4-week-old SHRSP were significantly greater than those of age-matched Wistar-Kyoto rats. Spironolactone prevented cardiac hypertrophy in adrenalectomized SHRSP. These results suggest that the rat heart produces aldosterone and that endogenous cardiac aldosterone may affect cardiac function and hypertrophy in hypertension in rats. PMID- 11040226 TI - Leptin attenuates cardiac contraction in rat ventricular myocytes. Role of NO. AB - Obesity is commonly associated with impaired myocardial contractile function. However, a direct link between these 2 states has not yet been established. There has been an indication that leptin, the product of the human obesity gene, may play a role in obesity-related metabolic and cardiovascular dysfunctions. The purpose of this study was to determine whether leptin exerts any direct cardiac contractile action that may contribute to altered myocardial function. Ventricular myocytes were isolated from adult male Sprague-Dawley rats. Contractile responses were evaluated by use of video-based edge detection. Contractile properties analyzed in cells electrically stimulated at 0.5 Hz included peak shortening, time to 90% peak shortening, time to 90% relengthening, and fluorescence intensity change. Leptin exhibited a dose-dependent inhibition in myocyte shortening and intracellular Ca(2+) change, with maximal inhibitions of 22.4% and 26.2%, respectively. Pretreatment with the NO synthase inhibitor N:(omega)-nitro-L-arginine methyl ester (L-NAME, 100 micromol/L) blocked leptin induced inhibition of both peak shortening and fluorescence intensity change. Leptin also stimulated NO synthase activity in a time- and concentration dependent manner, as reflected in the dose-related increase in NO accumulation in these cells. Addition of an NO donor (S-nitroso-N-acetyl-penicillamine [SNAP]) to the medium mimicked the effects of leptin administration. In summary, this study demonstrated a direct action of leptin on cardiomyocyte contraction, possibly through an increased NO production. These data suggest that leptin may play a role in obesity-related cardiac contractile dysfunction. PMID- 11040227 TI - Enhanced expression of angiotensin II type 2 receptor, inositol 1,4, 5 trisphosphate receptor, and protein kinase cepsilon during cardioprotection induced by angiotensin II type 2 receptor blockade. AB - We hypothesized that the cardioprotective effect of angiotensin II type 2 receptor (AT(2)R) blockade with PD 123,319 (PD) on the recovery of left ventricular (LV) mechanical function after ischemia/reperfusion (IR) in the isolated working rat heart is associated with the enhanced expression of AT(2)R protein and mRNA as well as an increase in inositol 1,4,5-trisphosphate type 2 receptor (IP(3)R) and protein kinase Cepsilon (PKCepsilon) proteins. We assessed AT(2)R, angiotensin II type 1 receptor (AT(1)R), IP(3)R, and PKCepsilon protein expression (Western blots) and AT(2)R mRNA levels (Northern blots) in myocardium from isolated working rat hearts that were subjected to global ischemia (30 minutes) followed by reperfusion (30 minutes). Groups of adult rat hearts (n=6) were exposed to no IR, no IR+PD (0.3 micromol/L), IR, and IR+PD. Compared with no IR and no IR+PD, IR decreased (P<0.05) functional recovery and AT(2)R mRNA and protein, as well as AT(1)R mRNA (not protein) and IP(3)R and PKCepsilon proteins. Compared with IR, PD+IR improved LV functional recovery (P<0.05) and markedly increased AT(2)R mRNA and protein (P<0.001). However, PD did not change AT(1)R mRNA or protein. More importantly, PD+IR markedly increased IP(3)R and PKCepsilon proteins. The downregulation of AT(2)R mRNA and protein with IR and their upregulation with PD indicate that the effects of PD are AT(2)R specific. The overall results suggest that the cardioprotective effect of acute PD treatment on LV functional recovery after IR in the isolated working rat heart is specifically due to AT(2)R blockade and is associated with enhanced downstream IP(3)R and PKCepsilon signaling. PMID- 11040229 TI - Transforming growth factor beta in hypertensives with cardiorenal damage. AB - We investigated whether a relationship exists between circulating transforming growth factor beta -1 (TGF-beta(1)), collagen type I metabolism, microalbuminuria, and left ventricular hypertrophy in essential hypertension and whether the ability of the angiotensin II type 1 receptor antagonist losartan to correct microalbuminuria and regress left ventricular hypertrophy in hypertensives is related to changes in TGF-beta(1) and collagen type I metabolism. The study was performed in 30 normotensive healthy controls and 30 patients with never-treated essential hypertension classified into 2 groups: those with microalbuminuria (urinary albumin excretion >30 and <300 mg/24 h) associated with left ventricular hypertrophy (left ventricular mass index >116 g/m(2) for men and >104 g/m(2) for women) (group B; n=17) and those without microalbuminuria or left ventricular hypertrophy (group A; n=13). The measurements were repeated in all patients after 6 months of treatment with losartan (50 mg once daily). The serum concentration of TGF-beta(1) was measured by a 2-site ELISA method, and the serum concentrations of carboxy-terminal propeptide of procollagen type I (a marker of collagen type I synthesis) and carboxy-terminal telopeptide of collagen type I (a marker of collagen type I degradation) were measured by specific radioimmunoassays. The duration of hypertension and baseline values of blood pressure were similar in the 2 groups of patients. No differences in serum TGF-beta(1), carboxy-terminal propeptide of procollagen type I, and carboxy-terminal telopeptide of collagen type I were found between normotensives and group A of hypertensives. Serum TGF-beta(1), carboxy-terminal propeptide of procollagen type I, and the ratio of carboxy terminal propeptide of procollagen type I to carboxy-terminal telopeptide of collagen type I were increased (P<0.05) in group B of hypertensives compared with group A of hypertensives and normotensives. No differences in carboxy-terminal telopeptide of collagen type I were found among the 3 groups of subjects. After treatment with losartan, microalbuminuria and left ventricular hypertrophy persisted in 6 patients (then considered nonresponders) and disappeared in 11 patients (then considered responders) from group B. Compared with nonresponders, responders exhibited similar control of blood pressure and higher (P<0.05) blockade of angiotensin II type 1 receptors (as assessed by a higher increase in plasma levels of angiotensin II). Whereas TGF-beta(1), carboxy-terminal propeptide of procollagen type I, and the ratio of carboxy-terminal propeptide of procollagen type I to carboxy-terminal telopeptide of collagen type I decreased (P<0.05) in responders, no changes in these parameters were observed in nonresponders. These findings show that an association exists between an excess of TGF-beta(1), stimulation of collagen type I synthesis, inhibition of collagen type I degradation, and cardiorenal damage in a group of patients with essential hypertension. In addition, our results suggest that the ability of losartan to blunt the synthesis of TGF-beta(1) and normalize collagen type I metabolism may contribute to protect the heart and the kidney in a fraction of patients with essential hypertension. PMID- 11040228 TI - Important role of angiotensin II-mediated c-Jun NH(2)-terminal kinase activation in cardiac hypertrophy in hypertensive rats. AB - In vitro studies on the role of the mitogen-activated protein (MAP) kinase family (extracellular signal-regulated kinase [ERK], c-Jun NH(2)-terminal kinase [JNK], and p38) in cardiac hypertrophic response have produced confusing and contradictory results. We examined the in vivo role of the angiotensin II type 1 (AT(1)) receptor in cardiac MAP kinase activities during both the onset and development of cardiac hypertrophy in stroke-prone spontaneously hypertensive rats (SHRSP). In both the acute and chronic phases of cardiac hypertrophy in SHRSP, cardiac JNK activities were significantly increased compared with those in normotensive rats, whereas there was no prominent increase in cardiac ERK or p38 activities in SHRSP. Losartan, an AT(1) receptor antagonist, prevented the onset of cardiac hypertrophy and regressed the progression of cardiac hypertrophy in SHRSP, being accompanied by the reduction of JNK activity and activator protein-1 (AP-1) activity in SHRSP. However, in spite of the normalization of blood pressure, hydralazine did not prevent or regress cardiac hypertrophy and did not decrease JNK or AP-1 activity in SHRSP. Inversely, hydralazine significantly increased the cardiac ERK activity in SHRSP by enhancing its phosphorylation. In conclusion, we have obtained the first evidence that the AT(1) receptor is involved in the enhanced cardiac JNK activity in both the onset and development of cardiac hypertrophy of hypertensive rats. We propose that JNK is involved in AT(1) receptor-mediated cardiac hypertrophy in vivo, in part mediated by the activation of AP-1. PMID- 11040230 TI - Beneficial renal and hemodynamic effects of omapatrilat in mild and severe heart failure. AB - Omapatrilat is a member of the new drug class of vasopeptidase inhibitors that may offer benefit in the treatment of heart failure (HF) through simultaneous inhibition of angiotensin-converting enzyme and neutral endopeptidase. We examined the effects of omapatrilat in a placebo-controlled crossover study using a pacing model of HF. Seven sheep were paced sequentially at 180 bpm (mild HF) and then 225 bpm (severe HF) for 7 days each. Omapatrilat (0.005 mg/kg) or vehicle was administered by intravenous bolus on days 4 to 7 of each paced period. Omapatrilat lowered mean arterial and left atrial pressure and increased cardiac output acutely and chronically in both mild and severe HF (P<0.01 for all). Plasma atrial and brain natriuretic peptide and cGMP levels were stable acutely (P=NS), while brain natriuretic peptide increased after repeated dosing in severe HF (P<0.05). Plasma renin activity rose, whereas angiotensin II and aldosterone levels fell after acute and repeated dosing in both states (P<0.01 for all). Omapatrilat increased urinary sodium excretion by day 7 in both mild and severe HF (P<0.05). Effective renal plasma flow and glomerular filtration rate increased or were stable after omapatrilat in mild and severe HF after both acute and repeated dosing. Omapatrilat exhibited pronounced acute and sustained beneficial hemodynamic and renal effects in both mild and severe heart failure. PMID- 11040231 TI - Ethnic differences in insulinemia and sympathetic tone as links between obesity and blood pressure. AB - Hyperinsulinemia and increased sympathetic nervous system (SNS) activity are thought to be pathophysiological links between obesity and hypertension. In the present study, we examined the relation among heart rate (HR), blood pressure (BP), and percent body fat (hydrodensitometry or DEXA), fasting plasma insulin concentration, and muscle sympathetic nerve activity (MSNA, microneurography) in male, normotensive whites (n=42) and Pima Indians (n=77). Pima Indians have a high prevalence of obesity and hyperinsulinemia but a relatively low prevalence of hypertension. Compared with whites, Pima Indian men had a higher percent body fat (28% versus 21%) and higher fasting insulin concentrations (210 versus 132 pmol/L) but lower MSNA (27 versus 33 bursts/min) (all P<0.001). In both ethnic groups, HR and BP were positively related to percent body fat and MSNA, and both were significant independent determinants of HR and BP in multiple regression analyses. However, MSNA was positively related to percent body fat and the fasting insulin concentration in whites (r=0.60 and r=0.47, both P<0.01) but not in Pima Indians (r=0.15 and r=0.03, NS) (P<0.01 for ethnic differences in the slope of the regression lines). These results confirm the physiological importance of the SNS in normal BP regulation but indicate that the roles of hyperinsulinemia and increased SNS activity as mediators for the relation between obesity and hypertension can differ between different ethnic groups. The lack of an increase in SNS activity with increasing adiposity and insulinemia in Pima Indians may contribute to the low prevalence of hypertension in this population. PMID- 11040232 TI - Adrenergic and reflex abnormalities in obesity-related hypertension. AB - Previous studies have shown that essential hypertension and obesity are both characterized by sympathetic activation coupled with a baroreflex impairment. The present study was aimed at determining the effects of the concomitant presence of the 2 above-mentioned conditions on sympathetic activity as well as on baroreflex cardiovascular control. In 14 normotensive lean subjects (aged 33. 5+/-2.2 years, body mass index 22.8+/-0.7 kg/m(2) [mean+/-SEM]), 16 normotensive obese subjects (body mass index 37.2+/-1.3 kg/m(2)), 13 lean hypertensive subjects (body mass index 24.0+/-0.8 kg/m(2)), and 16 obese hypertensive subjects (body mass index 37.5+/-1.3 kg/m(2)), all age-matched, we measured beat-to-beat arterial blood pressure (by Finapres device), heart rate (HR, by ECG), and postganglionic muscle sympathetic nerve activity (MSNA, by microneurography) at rest and during baroreceptor stimulation and deactivation induced by stepwise intravenous infusions of phenylephrine and nitroprusside, respectively. Blood pressure values were higher in lean hypertensive and obese hypertensive subjects than in normotensive lean and obese subjects. MSNA was significantly (P:<0.01) greater in obese normotensive subjects (49.1+/-3.0 bursts per 100 heart beats) and in lean hypertensive subjects (44.5+/-3.3 bursts per 100 heart beats) than in lean normotensive control subjects (32.2+/-2.5 bursts per 100 heart beats); a further increase was detectable in individuals with the concomitant presence of obesity and hypertension (62.1+/-3. 4 bursts per 100 heart beats). Furthermore, whereas in lean hypertensive subjects, only baroreflex control of HR was impaired, in obese normotensive subjects, both HR and MSNA baroreflex changes were attenuated, with a further attenuation being observed in obese hypertensive patients. Thus, the association between obesity and hypertension triggers a sympathetic activation and an impairment in baroreflex cardiovascular control that are greater in magnitude than those found in either of the above-mentioned abnormal conditions alone. PMID- 11040233 TI - Differentiated response of the sympathetic nervous system to angiotensin converting enzyme inhibition in hypertension. AB - Hypertension with renal artery stenosis is associated with both an activated renin-angiotensin system and elevated sympathetic activity. Therefore, in this condition it may be favorable to use a therapeutic modality that does not reflexly increase heart rate, renin secretion, and sympathetic nervous activity. The purpose of the present study was to assess overall, renal, and muscle sympathetic activity after short-term administration of an angiotensin-converting enzyme inhibitor (enalaprilat) and a nonspecific vasodilator (dihydralazine) to hypertensive patients with renal artery stenosis. Forty-eight patients undergoing a clinical investigation for renovascular hypertension were included in the study. An isotope dilution technique for assessing norepinephrine spillover was used to estimate overall and bilateral renal sympathetic nerve activity. In 11 patients simultaneous intraneural recordings of efferent muscle sympathetic nerve activity were performed. Thirty minutes after dihydralazine administration, mean arterial pressure fell by 15%, whereas plasma angiotensin II, muscle sympathetic nerve activity, heart rate, and total body norepinephrine spillover increased (P<0.05 for all). In contrast, after enalaprilat administration a fall in arterial pressure similar to that for dihydralazine was followed by decreased angiotensin II levels and unchanged muscle sympathetic nerve activity, heart rate, and total body norepinephrine spillover, whereas renal norepinephrine spillover increased by 44% (P<0.05). Acute blood pressure reduction by an angiotensin-converting enzyme inhibitor provokes a differentiated sympathetic response in patients with hypertension and renal artery stenosis, inasmuch that overall and muscle sympathetic reflex activation are blunted, whereas the reflex renal sympathetic response to blood pressure reduction is preserved. PMID- 11040234 TI - Development of hypertension induced by subpressor infusion of angiotensin II: role of sensory nerves. AB - Long-term administration of a subpressor dose of angiotensin II (Ang II) leads to pressor hyperresponsiveness and slow development of hypertension. Our preliminary data show that mRNA expression for calcitonin-gene related peptide in dorsal root ganglia was significantly increased by subpressor infusion of Ang II. To determine the role of sensory nerves in the development of hypertension induced by subpressor infusion of Ang II, newborn Wistar rats were given 50 mg/kg SC capsaicin on the 1st and 2nd days of life. After the weaning period, male rats were divided into 4 groups and subjected to the following treatments for 2 weeks: capsaicin+Ang II (150 ng. kg(-1). min(-1) SC by osmotic pumps, CAP-AII), capsaicin+vehicle (CAP), control+Ang II (CON-AII), and control+vehicle (CON). The results show that mean arterial pressure was significantly elevated in both Ang II-infused rats compared with non-Ang II-treated rats (P<0.05), and it was higher in CAP-AII than in CON-AII rats (P<0.05). The 24-hour urinary and sodium excretions were lower in CAP-AII than in CON-AII, CAP, and CON rats (P<0.05). These data demonstrated that sensory denervation exacerbates the development of hypertension and impairs renal excretory function when a subpressor dose of Ang II is given. These results indicate that activation of sensory nerves, either by Ang II or by other hormonal or hemodynamic factors, plays a compensatory role in promoting urine and sodium excretion and attenuating elevated blood pressure initiated by Ang II. PMID- 11040235 TI - Endogenous circulating sympatholytic factor in orthostatic intolerance. AB - Sympathotonic orthostatic hypotension (SOH) is an idiopathic syndrome characterized by tachycardia, hypotension, elevated plasma norepinephrine, and symptoms of orthostatic intolerance provoked by assumption of an upright posture. We studied a woman with severe progressive SOH with blood pressure unresponsive to the pressor effects of alpha(1)-adrenergic receptor (AR) agonists. We tested the hypothesis that a circulating factor in this patient interferes with vascular adrenergic neurotransmission. Preincubation of porcine pulmonary artery vessel rings with patient plasma produced a dose-dependent inhibition of vasoconstriction to phenylephrine in vitro, abolished vasoconstriction to direct electrical stimulation, and had no effect on nonadrenergic vasoconstrictive stimuli (endothelin-1), PGF-2alpha (or KCl). Preincubation of vessels with control plasma was devoid of these effects. SOH plasma inhibited the binding of an alpha(1)-selective antagonist radioligand ([(125)I]HEAT) to membrane fractions derived from porcine pulmonary artery vessel rings, rat liver, and cell lines selectively overexpressing human ARs of the alpha(1B) subtype but not other AR subtypes (alpha(1A) and alpha(1D)). We conclude that a factor in SOH plasma can selectively and irreversibly inhibit adrenergic ligand binding to alpha(1B) ARs. We propose that this factor contributes to a novel pathogenesis for SOH in this patient. This patient's syndrome represents a new disease entity, and her plasma may provide a unique tool for probing the selective functions of alpha(1)-ARs. PMID- 11040236 TI - Blockade of the renin-angiotensin and endothelin systems on progressive renal injury. AB - The renin-angiotensin system (RAS) and endothelin system may both play a role in the pathogenesis of progressive renal injury. The aims of the present study were 3-fold: first, to explore the possible benefits of dual blockade of the RAS with an ACE inhibitor and an angiotensin type 1(AT1) receptor antagonist; second, to examine the relative efficacy of endothelin A receptor antagonism (ETA-RA) compared with combined endothelin A/B receptor antagonism (ETA/B-RA); and third, to assess whether interruption of both RAS and endothelin system had any advantages over single-system blockade. Subtotally nephrectomized rats were studied as a model of progressive renal injury and randomly assigned to one of the following treatments for 12 weeks: perindopril (ACE inhibitor), irbesartan (AT1 receptor antagonist), BMS193884 (ETA-RA), bosentan (ETA/B-RA), and a combination of irbesartan with either perindopril or BMS193884. Treatment with irbesartan or perindopril was associated with an improved glomerular filtration rate and reductions in blood pressure, urinary protein excretion, glomerulosclerosis, and tubular injury in association with reduced gene expression of transforming growth factor-beta(1) and matrix protein type IV collagen. The combination of irbesartan with perindopril was associated with further reductions in blood pressure and urinary protein excretion. No beneficial effects of either BMS193884 or bosentan were noted. Furthermore, the addition of BMS193884 to irbesartan did not confer any additional benefits. These findings suggest that the RAS but not the endothelin system is a major mediator of progressive renal injury after renal mass reduction and that the combination of an AT1 receptor antagonist with an ACE inhibitor may have advantages over the single agent of RAS blocker treatment. PMID- 11040237 TI - Effect of angiotensin II blockade on renal injury in mineralocorticoid-salt hypertension. AB - Kidney function and structure were compared in control rats (group 1) and in 3 groups of rats made hypertensive by administration of aldosterone and saline for 8 weeks (groups 2, 3, and 4). Group 2 rats received only aldosterone and saline, while group 3 also received losartan and group 4 also received enalapril. Rats in all groups were subjected to uninephrectomy before beginning the experiment. Hypertension and proteinuria in rats given aldosterone and saline were not affected by losartan or enalapril (8-week values for blood pressure in mm Hg: 135+/-3 group 1, 193+/-4 group 2, 189+/-4 group 3, 189+/-5 group 4; P<0.05 groups 2, 3, and 4 versus 1; 8-week values for proteinuria in mg/d: 44+/-8 group 1, 278+/-34 group 2, 267+/-37 group 3, 289+/-36 group 4; P<0.05 groups 2, 3, and 4 versus 1). Vascular, glomerular, and tubulointerstitial injury accompanied hypertension and proteinuria at 8 weeks. Losartan and enalapril did not prevent vascular injury, which was characterized by thickening of arterial and arteriolar walls and by fibrinoid necrosis and thrombotic microangiopathy. Likewise, losartan and enalapril did not reduce the prevalence of glomerular segmental sclerosis (1+/-1% group 1, 10+/-2% group 2, 11+/-2% group 3, 13+/-2% group 4; P<0.05 groups 2, 3, and 4 versus 1) or limit tubulointerstitial injury as reflected by the volume fraction of the cortical interstitium (15+/-1% group 1, 20+/-1% group 2, 21+/-1% group 3, 21+/-1% group 4; P<0.05 groups 2, 3, and 4 versus 1). These findings suggest that local angiotensin II activity does not contribute to the development of renal injury in mineralocorticoid-salt hypertension. PMID- 11040238 TI - Renin-angiotensin system blockade improves endothelial dysfunction in hypertension. AB - Angiotensin-converting enzyme (ACE) inhibitor improves the impaired hyperpolarization and relaxation to acetylcholine (ACh) via endothelium-derived hyperpolarizing factor (EDHF) in arteries of spontaneously hypertensive rats (SHR). We tested whether the angiotensin type 1 (AT(1)) receptor antagonist also improves EDHF-mediated responses and whether the combined AT(1) receptor blockade and ACE inhibition exert any additional effects. SHR were treated with either AT(1) receptor antagonist TCV-116 (5 mg. kg(-1). d(-1)) (SHR-T), enalapril (40 mg. kg(-1). d(-1)) (SHR-E), or their combination (SHR-T&E) from 8 to 11 months of age. Age-matched, untreated SHR (SHR-C) and Wistar Kyoto (WKY) rats served as controls (n=8 to 12 in each group). Three treatments lowered blood pressure comparably. EDHF-mediated hyperpolarization to ACh in mesenteric arteries in the absence or presence of norepinephrine was significantly improved in all treated SHR. In addition, the hyperpolarization in the presence of norepinephrine was significantly greater in SHR-T&E than in SHR-E (ACh 10(-5) mol/L with norepinephrine: SHR-C -7; SHR-T -19; SHR-E -15; SHR-T&E -22; WKY -14 mV). EDHF mediated relaxation, assessed in the presence of indomethacin and N:(G)-nitro-L arginine, was markedly improved in all treated SHR. Hyperpolarization and relaxation to levcromakalim, a direct opener of ATP-sensitive K(+)-channel, were similar in all groups. These findings suggest that AT(1) receptor antagonists are as effective as ACE inhibitors in improving EDHF-mediated responses in SHR. The beneficial effects of the combined AT(1) receptor blockade and ACE inhibition appears to be for the most part similar to those of each intervention. PMID- 11040239 TI - Angiotensin-converting enzyme inhibitor prevents age-related endothelial dysfunction. AB - Vascular relaxation via endothelium-derived hyperpolarizing factor (EDHF) declines in association with aging and also with hypertension, and antihypertensive treatment improves the endothelial dysfunction connected with hypertension. We tested whether the angiotensin-converting enzyme inhibitor improves EDHF-mediated responses in normotensive rats, with special reference to the age-related process. Wistar-Kyoto rats (WKY) were treated with either 20 mg. kg(-1). d(-1) enalapril (WKY-E group) or a combination of 50 mg. kg(-1). d(-1) hydralazine and 7.5 mg. kg(-1). d(-1) hydrochlorothiazide (WKY-H group) from 9 to 12 months of age. Twelve-month-old WKY (WKY-O) and 3-month-old WKY (WKY-Y) served as controls (n=6 to 10 in each group). The 2 treatments lowered systolic blood pressure comparably. EDHF-mediated hyperpolarization to acetylcholine (ACh) in mesenteric arteries was significantly improved in WKY-E, but not in WKY-H, compared with WKY-O, and the hyperpolarization in WKY-E was comparable to that in WKY-Y (hyperpolarization to 10(-)(5) mol/L ACh in the presence of norepinephrine: WKY-O, -14+/-2 mV; WKY-E, -22+/-3 mV; WKY-H, -15+/-2 mV; and WKY-Y, -28+/-0 mV). EDHF-mediated relaxation, as assessed by relaxation to ACh in norepinephrine precontracted rings in the presence of indomethacin and NO synthase inhibitor, was also significantly improved in WKY-E, but not in WKY-H, to a level comparable to that in WKY-Y (maximum relaxation: WKY-O, 45+/-6%; WKY-E, 63+/-8%; WKY-H, 43+/ 4%; and WKY-Y, 72+/-4%). Hyperpolarization and relaxation to levcromakalim, an ATP-sensitive K(+) channel opener, were similar in all groups. These findings suggest that the angiotensin-converting enzyme inhibitor prevents the age-related decline in EDHF-mediated hyperpolarization and relaxation in normotensive rats, presumably through an inhibition of the renin-angiotensin system. PMID- 11040240 TI - Hemodynamic, hormone, and urinary effects of adrenomedullin infusion in essential hypertension. AB - We examined the effects of the vasodilator peptide adrenomedullin (AM) infused intravenously into subjects with essential hypertension. Eight men 39 to 58 years old with uncomplicated hypertension (147/96+/-5/3 mm Hg at baseline) were studied in a placebo-controlled, crossover design. Each subject received intravenous AM in a low and a high dose (2.9 and 5.8 pmol. kg(-1). min(-1) for 2 hours each) or vehicle-control (Hemaccel) infusion in a random order on day 4 of a controlled metabolic diet (80 mmol/d Na(+), 100 mmol/d K(+)). Plasma AM reached pathophysiological levels during infusion (18+/-4 pmol/L in low dose, 34+/-9 pmol/L in high dose) with a concurrent rise in plasma cAMP (+8.4+/-1.2 pmol/L, P:<0. 05 compared with control). Compared with control, high-dose AM increased peak heart rate (+17.8+/-2.3 bpm, P<0.01), lowered systolic (-24.6+/-0.9 mm Hg; P<0.01) and diastolic (-21.9+/-1.4 mm Hg; P<0.01) blood pressure, and increased cardiac output (+1.0+/-0. 1 L/min in low dose, +2.9+/-0.2 L/min in high dose; P<0.01 for both). Despite a rise in plasma renin activity during high dose (P<0.05), aldosterone levels did not alter. Plasma norepinephrine levels increased 1295+/-222 pmol/L (P<0.001) and epinephrine increased 74+/-15 pmol/L (P<0.05) with high-dose AM compared with control. AM had no significant effect on urine volume and sodium excretion. In subjects with essential hypertension, the intravenous infusion of AM to achieve pathophysiological levels produced significant falls in arterial pressure, increased heart rate and cardiac output, and stimulated the sympathetic system and renin release without concurrent increase in aldosterone. Urinary parameters were unaltered. Although AM has potent hemodynamic and neurohumoral effects in subjects with essential hypertension, the threshold for urinary actions is set higher. PMID- 11040241 TI - Differential control of systolic and diastolic blood pressure : factors associated with lack of blood pressure control in the community. AB - Data from the Third National Health and Nutrition Examination Survey, phase 2 (1991 to 1994), indicate that among hypertensive individuals in the United States, 53.6% are treated and only 27.4% are controlled to goal levels. We sought to determine whether poor hypertension control is due to lack of systolic or diastolic blood pressure control, or both. We studied Framingham Heart Study participants examined between 1990 and 1995 and determined rates of control to systolic goal (<140 mm Hg), diastolic goal (<90 mm Hg), or both (systolic <140 and diastolic <90 mm Hg). Of 1959 hypertensive subjects (mean age 66 years, 54% women), 32.7% were controlled to systolic goal, 82.9% were controlled to diastolic goal, and only 29.0% were controlled to both. Among the 1189 subjects who were receiving antihypertensive therapy (60.7% of all hypertensive subjects), 49.0% were controlled to systolic goal, 89.7% were controlled to diastolic goal, and only 47.8% were controlled to both. Thus, poor systolic blood pressure control was overwhelmingly responsible for poor rates of overall control to goal. Covariates associated with lack of systolic control in treated subjects included older age (OR for age 61 to 75 years, 2.43, 95% CI 1.79 to 3.29; OR for age >75 years, 4.34, 95% CI 3.10 to 6.09), left ventricular hypertrophy (OR 1.63, 95% CI 1.04 to 2.54), and obesity (OR for body mass index >/=30 versus <25 kg/m(2), 1.49, 95% CI 1.08 to 2.06). In this community-based sample of middle-aged and older subjects, overall rates of hypertension control were remarkably similar to those in the Third National Health and Nutrition Examination Survey. Poor blood pressure control was overwhelmingly due to lack of systolic control, even among treated subjects. Therefore, clinicians and policymakers should place greater emphasis on the achievement of goal systolic levels in all hypertensive patients, especially those who are older or obese or have target organ damage. PMID- 11040242 TI - Trends in antihypertensive drug therapy of ambulatory patients by US office-based physicians. AB - This study assessed trends from 1980 to 1995 in ambulatory patients' antihypertensive drug therapy by US office-based physicians for visits in which hypertension was the principal diagnosis and compared these trends with the respective guidelines given in 5 Joint National Committee (JNC) Reports on Detection, Evaluation, and Treatment of High Blood Pressure published around the same time period. Data from the National Center for Health Statistics' National Ambulatory Medical Care Surveys for 1980, 1985, 1990, and 1995 were used. From 1980 to 1995, there was no significant trend in the percentage of hypertension visits that did not mention any antihypertensive drug (20% to 27%). Further analyses focused on those hypertension visits in which at least 1 antihypertensive drug was used. Across the years, antihypertensive drug visits mentioning calcium channel blockers or ACE inhibitors significantly increased; those noting diuretics significantly decreased. However, in 1995, antihypertensive drug visits that included a diuretic and/or a beta-adrenergic blocker equalled 53%; these are the antihypertensive drug classes preferred by the JNC V. Physician antihypertensive drug prescribing was generally consistent with the basic antihypertensive drug guidelines of the JNC reports. PMID- 11040243 TI - Angiotensin II-induced hypertension: contribution of Ras GTPase/Mitogen-activated protein kinase and cytochrome P450 metabolites. AB - We reported that norepinephrine and angiotensin II (Ang II) activate the Ras/mitogen-activated protein (MAP) kinase pathway primarily through the generation of cytochrome P450 (CYP450) metabolites. The purpose of the present study was to determine the contribution of Ras and CYP450 to Ang II-dependent hypertension in rats. Infusion of Ang II (350 ng/min for 6 days) elevated mean arterial blood pressure (MABP) (171+/-3 mm Hg for Ang II versus 94+/-5 for vehicle group, P<0.05). Ras is activated on farnesylation by farnesyl protein transferase (FPT). When Ang II was infused in combination with FPT inhibitor FPT III (232 ng/min) or BMS-191563 (578 ng/min), the development of hypertension was attenuated (171+/-3 mm Hg for Ang II plus vehicle versus 134+/-5 mm Hg for Ang II plus FPT III and 116+/-6 mm Hg for Ang II plus BMS-191563, P<0.05). Treatment with the MAP kinase kinase inhibitor PD-98059 (5 mg SC) reduced MABP. The CYP450 inhibitor aminobenzotriazole (50 mg/kg) also diminished the development of Ang II induced hypertension to 113+/-8 mm Hg. The activities of Ras, MAP kinase, and CYP450 measured in the kidney were elevated in hypertensive animals. The infusion of FPT III, BMS-191563, or aminobenzotriazole reduced the elevation in Ras and MAP kinase activity. Morphological studies of the kidney showed that FPT III treatment ameliorated the arterial injury, vascular lesions, fibrinoid necrosis, focal hemorrhage, and hypertrophy of muscle walls observed in hypertensive animals. These data suggest that the activation of Ras and CYP450 contributes to the development of Ang II-dependent hypertension and associated vascular pathology. PMID- 11040244 TI - Cytochrome P450-dependent renal arachidonic acid metabolism in desoxycorticosterone acetate-salt hypertensive mice. AB - Cytochrome P450 (P450)-dependent arachidonic acid metabolites may act as mediators in the regulation of vascular tone and renal function. We studied arachidonic acid hydroxylase activities in renal microsomes from normotensive NMRI mice, desoxycorticosterone acetate (DOCA)-salt hypertensive mice, and DOCA salt mice treated with either lovastatin or bezafibrate, both of which improve hemodynamics in this model. Control renal microsomes had arachidonic acid hydroxylase activities of 175+/-12 pmol. min(-1). mg(-1). The metabolites formed were 20- and 19-hydroxyarachidonic acid, representing approximately 80% and approximately 20% of the total hydroxylation. Treatment with DOCA-salt resulted in significantly decreased hydroxylase activities (to 84+/-4 pmol. min(-1). mg( 1)) of the total microsomal P450 content and a decrease in immunodetectable Cyp4a proteins. Lovastatin had no effect on these variables, whereas bezafibrate increased arachidonic acid hydroxylase activities to 163+/-12 pmol. min(-1). mg( 1). In situ hybridization with probes for Cyp4a-10, 12, and 14 revealed that Cyp4a-14 was the P450 isoform most strongly induced by bezafibrate. The expression was concentrated in the cortical medullary junction and was localized predominantly in the proximal tubules. In conclusion, these results suggest that the capacity to produce 20-hydroxyarachidonic acid is impaired in the kidneys of DOCA-salt hypertensive mice. Furthermore, bezafibrate may ameliorate hemodynamics in this model by restoring P450-dependent arachidonic acid hydroxylase activities. Lovastatin, on the other hand, exerts its effects via P450 independent mechanisms. PMID- 11040245 TI - Angiotensin II stimulates extracellular signal-regulated kinase activity in intact pressurized rat mesenteric resistance arteries. AB - The activation of extracellular signal-regulated kinases 1/2 (ERK1/2) was assessed in isolated rat mesenteric resistance arteries (200-micrometer diameter) in a pressure myograph and stimulated for 5 minutes by angiotensin II (Ang II, 0.1 micromol/L) with a pressure of 70 mm Hg. ERK1/2 activity was measured by using an in-gel assay, and ERK1/2 phosphorylation was measured by Western blot analysis with use of a phospho-specific ERK1/2 antibody. Ang II (0.1 micromol/L) induced contraction (28% of phenylephrine contraction, 10 micromol/L). ERK kinase inhibitor PD98059 (10 micromol/L) attenuated this contraction by 36% but not that to phenylephrine or K(+) (60 mmol/L). In unpressurized arteries, Ang II increased ERK1/2 activity by 26%, and pressure (70 mm Hg) itself increased ERK1/2 activity by 72%. Ang II and pressure together acted synergistically, increasing ERK1/2 activity by 264%. Thus, in pressurized vessels, Ang II (0.1 micromol/L) increased ERK1/2 activity by 112%, calculated as [(364/172)-1]x100, which was confirmed by a measured 72% increase in ERK1/2 phosphorylation. Ang II type 1 receptor blockade by candesartan (10 micromol/L) abolished the Ang II-induced increase in ERK1/2 activity, but Ang II type 2 receptor blockade (PD123319, 10 micromol/L) did not. The Ang II-induced increase in ERK1/2 activity was inhibited by protein kinase C inhibitors Ro-31-8220 (1 micromol/L) and Go-6976 (300 nmol/L) and tyrosine kinase inhibitors genistein (1 micromol/L, general) and herbimycin A (1 micromol/L, c-Src family). The present findings show for the first time in intact resistance arteries that ERK1/2 activation is rapidly regulated by Ang II, is synergistic with pressure, and is involved in contraction. The ERK1/2 signaling pathway apparently includes upstream protein kinase C and c-Src. PMID- 11040247 TI - Colin johnston celebration PMID- 11040246 TI - Lacidipine and blood pressure variability in diabetic hypertensive patients. AB - The aim of our study was to assess the effects of lacidipine, a long-acting calcium antagonist, on 24-hour average blood pressure, blood pressure variability, and baroreflex sensitivity. In 10 mildly to moderately hypertensive patients with type II diabetes mellitus (aged 18 to 65 years), 24-hour ambulatory blood pressure was continuously monitored noninvasively (Portapres device) after a 3-week pretreatment with placebo and a subsequent 4-week once daily lacidipine (4 mg) or placebo treatment (double-blind crossover design). Systolic blood pressure, diastolic blood pressure, and heart rate means were computed each hour for 24 hours (day and night) at the end of each treatment period. Similar assessments were also made for blood pressure and heart rate variability (standard deviation and variation coefficient) and for 24-hour baroreflex sensitivity, which was quantified (1) in the time domain by the slope of the spontaneous sequences characterized by progressive increases or reductions of systolic blood pressure and RR interval and (2) in the frequency domain by the squared ratio of RR interval and systolic blood pressure spectral power approximately 0.1 and 0.3 Hz over the 24 hours. Compared with placebo, lacidipine reduced the 24-hour, daytime, and nighttime systolic and diastolic blood pressure (P<0.05) with no significant change in heart rate. It also reduced 24-hour, daytime, and nighttime standard deviation (-19.6%, -14.4%, and -24.0%, respectively; P<0.05) and their variation coefficient. The 24-hour average slope of all sequences (7.7+/-1.7 ms/mm Hg) seen during placebo was significantly increased by lacidipine (8.7+/-1.8 ms/mm Hg, P<0.01), with a significant increase being obtained also for the 24-hour average alpha coefficient at 0.1 Hz (from 5.7+/-1.5 to 6.4+/-1.3 ms/mm Hg, P<0.01). Thus, in diabetic hypertensive patients, lacidipine reduced not only 24-hour blood pressure means but also blood pressure variability. This reduction was accompanied by an improvement of baroreflex sensitivity. Computer analysis of beat-to-beat 24-hour noninvasive blood pressure monitoring may offer valuable information about the effects of antihypertensive drugs on hemodynamic and autonomic parameters in daily life. PMID- 11040248 TI - Honoring Colin Johnston. PMID- 11040250 TI - Hypothesis regarding the pathophysiological role of alternative pathways of angiotensin II formation in atherosclerosis. AB - The renin-angiotensin system has been studied and recognized as one of the major blood pressure-regulating systems for the past century. In the last quarter century, however, many alternative pathways of angiotensin II formation have been found, and among them, chymase has been a focus of interest because of its specificity and potency in the human cardiovascular system. Chymase evidently is not involved in functional regulation of blood pressure at least in the short term, but evidence is accumulating that it may be involved in structural remodeling of the cardiovascular system. We found increased vascular chymase activity in atherosclerotic lesions of the human aorta as well as in cardiac remodeling after myocardial infarction. We found a significant positive correlation between serum total or LDL cholesterol levels and arterial chymase dependent angiotensin II-forming activity in patients who were undergoing coronary artery bypass operation, suggesting that high serum cholesterol may trigger upregulation of vascular chymase and facilitate the development of atherosclerosis. This hypothesis was tested in Syrian hamsters fed a high cholesterol diet containing 0.5% cholesterol: A marked lipid deposition in the aortic cusp developed and the plasma cholesterol levels were positively correlated with aortic chymase activity. An orally active nonpeptide chymase inhibitor almost canceled this lipid deposition. These clinical and experimental data indicated an association between cholesterol and vascular chymase upregulation that may facilitate the development of atherosclerosis. PMID- 11040249 TI - Proximal sodium reabsorption: An independent determinant of blood pressure response to salt. AB - The purpose of this study was to evaluate the contribution of renal sodium handling by the proximal tubule as an independent determinant of blood pressure responsiveness to salt in hypertension. We measured blood pressure (BP), renal hemodynamics, and segmental renal sodium handling (with lithium used as a marker of proximal sodium reabsorption) in 38 hypertensive patients and 27 normotensive subjects (15 young and 12 age-matched) on a high and low sodium diet. In control subjects, changing the diet from a low to a high sodium content resulted in no change in BP and increases in glomerular filtration rate (P<0.05), renal plasma flow (P<0.05), and fractional excretion of lithium (FE(Li), P<0.01). In hypertensive patients, comparable variations of sodium intake induced an increase in BP with no change in renal hemodynamics and proximal sodium reabsorption. When analyzed by tertiles of their BP response to salt, salt-insensitive hypertensive patients of the first tertile disclosed a pattern of adaptation of proximal sodium reabsorption comparable to that of control subjects, whereas the most salt sensitive patients of the third tertile had an inverse pattern with a high FE(Li) on low salt and a lower FE(Li) on high salt, suggesting an inappropriate modulation of proximal sodium reabsorption. The BP response to salt correlated positively with age (r=0.34, P=0.036) and negatively with the changes in FE(Li) (r=-0.37, P=0.029). In a multivariate analysis, the changes in FE(Li) were significantly and independently associated with the salt-induced changes in BP. These results suggest that proximal sodium reabsorption is an independent determinant of the BP response to salt in hypertension. PMID- 11040251 TI - Nifedipine prevents changes in nitric oxide synthase mRNA levels induced by cyclosporine. AB - Cyclosporine toxicity mainly affects kidney and liver function. We have previously shown that cyclosporine nephrotoxicity alters kidney nitric oxide synthase mRNA pattern of expression. To determine if nitric oxide synthase expression changes are mediated directly by cyclosporine or by secondary hemodynamic alterations induced by cyclosporine, we evaluated if these effects are tissue specific and if nifedipine-induced vasodilation prevents these alterations. Uninephrectomized Wistar rats treated for 7 days with olive oil, cyclosporine (30 mg/kg), nifedipine (3 mg/kg), and nifedipine+cyclosporine were studied. In vehicle and cyclosporine groups, the gene expression of the neuronal, inducible, and endothelial nitric oxide synthases in cerebellum, heart, intestine, liver, renal cortex, and medulla was evaluated. The administration of cyclosporine was associated with nephrotoxicity and hepatotoxicity, increased endothelial nitric oxide synthase mRNA levels in renal cortex and liver, and a decrease in inducible nitric oxide synthase and neuronal nitric oxide synthase in renal medulla. The mRNA levels of the 3 nitric oxide synthase isoforms were not affected in any other tissue. Nifedipine did not alter nitric oxide synthase expression in the control group but prevented changes associated with cyclosporine. These results suggest that cyclosporine-induced changes in the pattern of expression of the nitric oxide synthases may be secondary to its hemodynamic effects. PMID- 11040252 TI - Renovascular hypertension: structural changes in the renal vasculature. AB - Experimental narrowing of the main renal artery to produce hypertension increases the aorta-glomerular capillary pressure difference and vascular resistance. This article examines the hypothesis that hypertension also may be caused by structural changes that narrow intrarenal blood vessels, similarly increasing preglomerular vascular resistance and the aortic-glomerular capillary pressure gradient. There is evidence of both wall hypertrophy and lumen narrowing of the preglomerular arteries in spontaneously hypertensive rats, with increased preglomerular resistance and aortic-glomerular capillary pressure difference. We have also attempted to induce structural changes in renal-preglomerular vessels experimentally by infusing angiotensin II at low doses (0.5 to 4.5 ng/kg per minute) into the renal artery of Sprague-Dawley rats and greyhound dogs for up to 4 weeks. This angiotensin II infusion produced apparent dose-related effects on preglomerular vessel structure and hypertension. The possibility that hypertension may be induced by structural changes in preglomerular resistance vessel walls, by simulation of the hemodynamic effects of main renal artery stenosis, deserves further investigation. PMID- 11040253 TI - The Council for High Blood Pressure Research is pleased to present: The Novartis Award for Hypertension Research. PMID- 11040254 TI - Gene expression and synaptic plasticity in the auditory forebrain of songbirds. PMID- 11040255 TI - Revisiting the maturation of medial temporal lobe memory functions in primates. PMID- 11040256 TI - Using pavlovian higher-order conditioning paradigms to investigate the neural substrates of emotional learning and memory. AB - In first-order Pavlovian conditioning, learning is acquired by pairing a conditioned stimulus (CS) with an intrinsically motivating unconditioned stimulus (US; e.g., food or shock). In higher-order Pavlovian conditioning (sensory preconditioning and second-order conditioning), the CS is paired with a stimulus that has motivational value that is acquired rather than intrinsic. This review describes some of the ways higher-order conditioning paradigms can be used to elucidate substrates of learning and memory, primarily focusing on fear conditioning. First-order conditioning, second-order conditioning, and sensory preconditioning allow for the controlled demonstration of three distinct forms of memory, the neural substrates of which can thus be analyzed. Higher-order conditioning phenomena allow one to distinguish more precisely between processes involved in transmission of sensory or motor information and processes involved in the plasticity underlying learning. Finally, higher-order conditioning paradigms may also allow one to distinguish between processes involved in behavioral expression of memory retrieval versus processes involved in memory retrieval itself. PMID- 11040257 TI - Parallel acquisition of awareness and trace eyeblink classical conditioning. AB - Trace eyeblink conditioning (with a trace interval >/=500 msec) depends on the integrity of the hippocampus and requires that participants develop awareness of the stimulus contingencies (i.e., awareness that the conditioned stimulus [CS] predicts the unconditioned stimulus [US]). Previous investigations of the relationship between trace eyeblink conditioning and awareness of the stimulus contingencies have manipulated awareness or have assessed awareness at fixed intervals during and after the conditioning session. In this study, we tracked the development of knowledge about the stimulus contingencies trial by trial by asking participants to try to predict either the onset of the US or the onset of their eyeblinks during differential trace eyeblink conditioning. Asking participants to predict their eyeblinks inhibited both the acquisition of awareness and eyeblink conditioning. In contrast, asking participants to predict the onset of the US promoted awareness and facilitated conditioning. Acquisition of knowledge about the stimulus contingencies and acquisition of differential trace eyeblink conditioning developed approximately in parallel (i.e., concurrently). PMID- 11040258 TI - Intact visual perceptual discrimination in humans in the absence of perirhinal cortex. AB - While the role of the perirhinal cortex in declarative memory has been well established, it has been unclear whether the perirhinal cortex might serve an additional nonmnemonic role in visual perception. Evidence that the perirhinal cortex might be important for visual perception comes from a recent report that monkeys with perirhinal cortical lesions are impaired on difficult (but not on simple) visual discrimination tasks. We administered these same tasks to nine amnesic patients, including three severely impaired patients with complete damage to perirhinal cortex bilaterally (E.P., G.P., and G.T.). The patients performed all tasks as well as controls. We suggest that the function of perirhinal cortex as well as antero-lateral temporal cortex may differ between humans and monkeys. PMID- 11040259 TI - Varying intertrial interval reveals temporally defined memory deficits and enhancements in NTAN1-deficient mice. AB - The N-end rule is one ubiquitin-proteolytic pathway that relates the in vivo half life of a protein to the identity of its N-terminal residue. NTAN1 deamidates N terminal asparagine to aspartate, which is conjugated to arginine by ATE1. An N terminal arginine-bearing substrate protein is recognized, ubiquitylated by UBR1/E3alpha, and subsequently degraded by 26S proteasomes. Previous research showed that NTAN1-deficient mice exhibited impaired long-term memory in the Lashley III maze. Therefore, a series of studies, designed to assess the role of NTAN1 in short- and intermediate-term memory processes, was undertaken. Two hundred sixty mice (126 -/-; 134 +/ +) received Lashley III maze training with intertrial intervals ranging from 2-180 min. Results indicated that inactivation of NTAN1 amidase differentially affects short-, intermediate-, and long-term memory. PMID- 11040260 TI - Effects of the selective M1 muscarinic receptor antagonist dicyclomine on emotional memory. AB - The nonselective muscarinic antagonist scopolamine is known to impair the acquisition of some learning tasks such as inhibitory avoidance. There has been recent research into the effects of this drug in contextual fear conditioning and tone fear conditioning paradigms. The purpose of the present study was to assess the role of the selective M1 muscarinic antagonist dicyclomine in these paradigms and in the inhibitory avoidance test. Rats were administered different doses of dicyclomine or saline 30 min before acquisition training. The animals were tested 24 hr later, and it was observed that 16 mg/kg of dicyclomine impaired both contextual fear conditioning and inhibitory avoidance. However, dicyclomine (up to 64 mg/kg) did not affect tone fear conditioning. These results suggest that the selective M1 muscarinic antagonist dicyclomine differentially affects aversively motivated tasks known to be dependent on hippocampal integrity (such as contextual fear conditioning and inhibitory avoidance) but does not affect similar hippocampus-independent tasks. PMID- 11040261 TI - Increasing acetylcholine levels in the hippocampus or entorhinal cortex reverses the impairing effects of septal GABA receptor activation on spontaneous alternation. AB - Intra-septal infusions of the gamma-aminobutyric acid (GABA) agonist muscimol impair learning and memory in a variety of tasks. This experiment determined whether hippocampal or entorhinal infusions of the acetylcholinesterase inhibitor physostigmine would reverse such impairing effects on spontaneous alternation performance, a measure of spatial working memory. Male Sprague-Dawley rats were given intra-septal infusions of vehicle or muscimol (1 nmole/0.5 microL) combined with unilateral intra-hippocampal or intra-entorhinal infusions of vehicle or physostigmine (10 microg/microL for the hippocampus; 7.5 microg/microL or 1.875 microg/0.25 microL for the entorhinal cortex). Fifteen minutes later, spontaneous alternation performance was assessed. The results indicated that intra-septal infusions of muscimol significantly decreased percentage-of-alternation scores, whereas intra-hippocampal or intra-entorhinal infusions of physostigmine had no effect. More importantly, intra-hippocampal or intra-entorhinal infusions of physostigmine, at doses that did not influence performance when administered alone, completely reversed the impairing effects of the muscimol infusions. These findings indicate that increasing cholinergic levels in the hippocampus or entorhinal cortex is sufficient to reverse the impairing effects of septal GABA receptor activation and support the hypothesis that the impairing effects of septal GABAergic activity involve cholinergic processes in the hippocampus and the entorhinal cortex. PMID- 11040262 TI - PACAP-38 enhances excitatory synaptic transmission in the rat hippocampal CA1 region. AB - Specific receptors for pituitary adenylate cyclase-activating polypeptide (PACAP), a novel peptide with neuroregulatory and neurotrophic functions, have been identified recently in different brain regions, including the hippocampus. In this study, we examined the effects of PACAP-38 on the excitatory postsynaptic field potentials (fEPSPs) evoked at the Schaffer collateral-CA1 synapses. Brief bath application of PACAP-38 (0.05 nM) induced a long-lasting facilitation of the basal transmission. Enhancement of this response was occluded in part by previous high-frequency-induced long-term potentiation (LTP). PACAP-38 did not significantly alter the paired-pulse facilitation (PPF). PACAP-38 has been shown to have a presynaptic effect on the septohippocampal cholinergic terminals, which results in an increase in basal acetylcholine (ACh) release. To assess whether the PACAP-38 enhancement of CA1 synapses was related to the activation of the cholinergic system we examined the effect of this peptide in the presence of atropine, a muscarinic receptor antagonist. The enhancement of the fEPSPs by PACAP-38 was blocked by bath application of atropine. These results show that PACAP-38 induces facilitation of hippocampal synaptic transmission through activation of the cholinergic system via the muscarinic receptors. PMID- 11040263 TI - Synaptic plasticity in hippocampal CA1 neurons of mice lacking type 1 inositol 1,4,5-trisphosphate receptors. AB - In hippocampal CA1 neurons of wild-type mice, delivery of a standard tetanus (100 pulses at 100 Hz) or a train of low-frequency stimuli (LFS; 1000 pulses at 1 Hz) to a naive input pathway induces, respectively, long-term potentiation (LTP) or long-term depression (LTD) of responses, and delivery of LFS 60 min after tetanus results in reversal of LTP (depotentiation, DP), while LFS applied 60 min before tetanus suppresses LTP induction (LTP suppression). To evaluate the role of the type 1 inositol-1,4,5-trisphosphate receptor (IP3R1) in hippocampal synaptic plasticity, we studied LTP, LTD, DP, and LTP suppression of the field excitatory postsynaptic potentials (EPSPs) in the CA1 neurons of mice lacking the IP3R1. No differences were seen between mutant and wild-type mice in terms of the mean magnitude of the LTP or LTD induced by a standard tetanus or LFS. However, the mean magnitude of the LTP induced by a short tetanus (10 pulses at 100 Hz) was significantly greater in mutant mice than in wild-type mice. In addition, DP or LTP suppression was attenuated in the mutant mice, the mean magnitude of the responses after delivery of LFS or tetanus being significantly greater than in wild-type mice. These results suggest that, in hippocampal CA1 neurons, the IP3R1 is involved in LTP, DP, and LTP suppression but is not essential for LTD. The facilitation of LTP induction and attenuation of DP and LTP suppression seen in mice lacking the IP3R1 indicates that this receptor plays an important role in blocking synaptic potentiation in hippocampal CA1 neurons. PMID- 11040264 TI - Input-specific immunolocalization of differentially phosphorylated Kv4.2 in the mouse brain. AB - Voltage-gated A-type potassium channels such as Kv4.2 regulate generation of action potentials and are localized abundantly in the hippocampus and striatum. Phosphorylation consensus sites for various kinases exist within the sequence of the potassium channel subunit Kv4.2, including consensus sites for extracellular signal-regulated kinase/mitogen activated protein kinase (ERK/MAPK), protein kinase A (PKA), protein kinase C (PKC), and calcium/calmodulin-dependent kinase II (CaMKII), and kinase assays have shown that particular amino acids of the consensus sites are bonafide phosphorylation sites in vitro. We have developed antibodies recognizing Kv4.2 triply phosphorylated at the three ERK sites as well as two antibodies recognizing singly phosphorylated Kv4.2 channels at the PKA sites (one amino-terminal and one carboxy-terminal). In the present study, we report the development of reliable immunohistochemistry protocols to study the localization of these phosphorylated versions of Kv4.2, as well as total Kv4.2 in the mouse brain. A general description of the areas highlighted by these antibodies includes the hippocampus, amygdala, cortex, and cerebellum. Such areas display robust synaptic plasticity and have been implicated in spatial, associative, and motor learning. Interestingly, in the hippocampus, the antibodies to differentially phosphorylated Kv4.2 channels localize to specific afferent pathways, indicating that the Kv4.2 phosphorylation state may be input specific. For example, the stratum lacunosum moleculare, which receives inputs from the entorhinal cortex via the perforant pathway, displays relatively little ERK-phosphorylated Kv4.2 or PKA carboxy-terminal-phosphorylated Kv4.2. However, this same layer is highlighted by antibodies that recognize Kv4.2 that has been phosphorylated by PKA at the amino terminus. Similarly, of the three antibodies tested, the soma of CA3 neurons are primarily recognized by the ERK triply phosphorylated Kv4.2 antibody, and the mossy fiber inputs to CA3 are primarily recognized by the carboxy-terminal PKA-phosphorylated Kv4.2. This differential phosphorylation is particularly interesting in two contexts. First, phosphorylation may be serving as a mechanism for targeting. For example, the amino-terminal PKA phosphorylation may be acting as a tag for a discrete pool of Kv4.2 to enter stratum lacunosum moleculare. Second, as phosphorylation may regulate channel biophysical properties, differential phosphorylation of Kv4.2 in the dendrites of pyramidal neurons may confer unique biophysical properties upon particular dendritic input layers. PMID- 11040265 TI - Role of hippocampal signaling pathways in long-term memory formation of a nonassociative learning task in the rat. AB - Long-term habituation to a novel environment is one of the most elementary forms of nonassociative learning. Here we studied the effect of pre- or posttraining intrahippocampal administration of drugs acting on specific molecular targets on the retention of habituation to a 5-min exposure to an open field measured 24 h later. We also determined whether the exposure to a novel environment resulted in the activation of the same intracellular signaling cascades previously shown to be activated during hippocampal-dependent associative learning. The immediate posttraining bilateral infusion of CNQX (1 microg/side), an AMPA/kainate glutamate receptor antagonist, or of muscimol (0.03 microg/side), a GABA(A) receptor agonist, into the CA1 region of the dorsal hippocampus impaired long term memory of habituation. The NMDA receptor antagonist AP5 (5 microg/side) impaired habituation when infused 15 min before, but not when infused immediately after, the 5-min training session. In addition, KN-62 (3.6 ng/side), an inhibitor of calcium calmodulin-dependent protein kinase II (CaMKII), was amnesic when infused 15 min before or immediately and 3 h after training. In contrast, the cAMP-dependent protein kinase (PKA) inhibitor Rp-cAMPS, the mitogen-activated protein kinase kinase (MAPKK) inhibitor PD098059, and the protein synthesis inhibitor anisomycin, at doses that fully block memory formation of inhibitory avoidance learning, did not affect habituation to a novel environment. The detection of spatial novelty is associated with a sequential activation of PKA, ERKs (p44 and p42 MAPKs) and CaMKII and the phosphorylation of c-AMP responsive element-binding protein (CREB) in the hippocampus. These findings suggest that memory formation of spatial habituation depends on the functional integrity of NMDA and AMPA/kainate receptors and CaMKII activity in the CA1 region of the hippocampus and that the detection of spatial novelty is accompanied by the activation of at least three different hippocampal protein kinase signaling cascades. PMID- 11040266 TI - A cGMP-dependent protein kinase gene, foraging, modifies habituation-like response decrement of the giant fiber escape circuit in Drosophila. AB - The Drosophila giant fiber jump-and-flight escape response is a model for genetic analysis of both the physiology and the plasticity of a sensorimotor behavioral pathway. We previously established the electrically induced giant fiber response in intact tethered flies as a model for habituation, a form of nonassociative learning. Here, we show that the rate of stimulus-dependent response decrement of this neural pathway in a habituation protocol is correlated with PKG (cGMP Dependent Protein Kinase) activity and foraging behavior. We assayed response decrement for natural and mutant rover and sitter alleles of the foraging (for) gene that encodes a Drosophila PKG. Rover larvae and adults, which have higher PKG activities, travel significantly farther while foraging than sitters with lower PKG activities. Response decrement was most rapid in genotypes previously shown to have low PKG activities and sitter-like foraging behavior. We also found differences in spontaneous recovery (the reversal of response decrement during a rest from stimulation) and a dishabituation-like phenomenon (the reversal of response decrement evoked by a novel stimulus). This electrophysiological study in an intact animal preparation provides one of the first direct demonstrations that PKG can affect plasticity in a simple learning paradigm. It increases our understanding of the complex interplay of factors that can modulate the sensitivity of the giant fiber escape response, and it defines a new adult-stage phenotype of the foraging locus. Finally, these results show that behaviorally relevant neural plasticity in an identified circuit can be influenced by a single locus genetic polymorphism existing in a natural population of Drosophila. PMID- 11040268 TI - Role of a striatal slowly inactivating potassium current in short-term facilitation of corticostriatal inputs: a computer simulation study. AB - Striatal output neurons (SONs) integrate glutamatergic synaptic inputs originating from the cerebral cortex. In vivo electrophysiological data have shown that a prior depolarization of SONs induced a short-term (beclomethasone (EC(50) 51+/-19 nM)>dexamethasone (EC(50) 303+/-40 nM). Hydrocortisone, prednisolone, and prednisone (up to 1 microM) did not induce any significant increase in eosinophil apoptosis. The apoptosis promoting effects of glucocorticoids on eosinophils were reversed by an antagonist of glucocorticoid receptor mifepristone. The survival-prolonging effect of tumor necrosis factor (TNF)-alpha was reversed by dexamethasone and fluticasone (1 microM). In contrast, fluticasone, and dexamethasone (1 microM) did not reverse the survival-prolonging effects of interleukins-3 and -5 or granulocyte-macrophage colony-stimulating factor (GM-CSF). The results suggest that fluticasone and budesonide induce eosinophil apoptosis at clinically achievable drug concentrations via an effect on glucocorticoid receptor. PMID- 11040339 TI - CV-2619 protects cultured astrocytes against reperfusion injury via nerve growth factor production. AB - In this study, we examined the effect of the neuroprotective agent 2, 3-dimethoxy 5-methyl-6-(10-hydroxydecyl)-1,4-benzoquinone (CV-2619) on reperfusion injury in cultured rat astrocytes after exposure to hydrogen peroxide (H(2)O(2))-containing medium. CV-2619 (10 nM to 10 microM) significantly attenuated the reperfusion induced decrease in cell viability. The compound showed an anti-apoptotic effect in this astrocyte injury model. Antioxidants such as ascorbic acid, alpha tocopherol and reduced glutathione also inhibited H(2)O(2) exposure-induced cytotoxicity. CV-2619 did not affect the levels of reactive oxygen species, but it increased nerve growth factor (NGF) production. The effect of CV-2619 on H(2)O(2) exposure-induced cytotoxicity was blocked by cycloheximide and anti-NGF antibody. The protective effect of CV-2619 was antagonized by the mitogen activated protein (MAP)/extracellular signal-regulated kinase (ERK) kinase inhibitor 2'-amino-3'-methoxyflavone and the phosphatidylinositol-3 kinase inhibitor wortmannin. These findings suggest that the effect of CV-2619 is mediated at least partly by NGF production in astrocytes and that ERK and phosphatidylinositol-3 kinases play a role in the downstream mechanism. PMID- 11040340 TI - Effects of fluoroquinolones on HERG currents. AB - We have investigated the effects of four fluoroquinolones on the human ether-a-go go-related gene (HERG) mediated K(+) currents to evaluate their potential to induce QT-prolongation. HERG currents were measured from stably transfected Chinese hamster ovary (CHO) cells by means of the patch-clamp technique. Bath application of sparfloxacin, moxifloxacin and grepafloxacin produced an inhibition of HERG outward currents at -40 mV with EC(50) of 13.5+/-0.8, 41. 2+/ 2.0 and 37.5+/-3.3 microg/ml, respectively. Current inhibitions were reversible after washout of the compounds. By contrast, ciprofloxacin at concentrations of up to 100 microg/ml did not effect HERG outward currents. PMID- 11040341 TI - Lack of tolerance to motor stimulant effects of a selective adenosine A(2A) receptor antagonist. AB - It is well known that tolerance develops to the actions of caffeine, which acts as an antagonist on adenosine A(1) and A(2A) receptors. Since selective adenosine A(2A) antagonists have been proposed as adjuncts to 3,4-dihydroxyphenylalanine (L DOPA) therapy in Parkinson's disease we wanted to examine if tolerance also develops to the selective A(2A) receptor antagonist 5-amino-7-(2-phenylethyl)-2 (2-furyl)-pyrazolo-[4,3-e]-1,2, 4-triazolo [1,5-c]pyrimidine (SCH 58261). SCH 58261 (0.1 and 7.5 mg/kg) increased basal locomotion and the motor stimulation afforded by apomorphine. Neither effect was subject to tolerance following long term treatment with the same doses given intraperitoneally twice daily. There were no adaptive changes in A(1) and A(2A) adenosine receptors or their corresponding messenger RNA or in dopamine D(1) or D(2) receptors. These results demonstrate that the tolerance that develops to caffeine is not secondary to its inhibition of adenosine A(2A) receptors. The results also offer hope that long term treatment with an adenosine A(2A) receptor antagonist may be possible in man. PMID- 11040342 TI - Influence of stimulation on Ca(2+) recruitment triggering [3H]acetylcholine release from the rat motor-nerve endings. AB - The influence of rat phrenic nerve stimulation frequency (5-50 Hz) and of pulse duration (0.04-1 ms) on Ca(2+) mobilization triggering [3H]acetylcholine release was investigated. The P-type voltage-dependent Ca(2+) channel (VDCC) blocker, omega-agatoxin IVA (100 nM), decreased [3H]acetylcholine release evoked by pulses of 0. 04-ms duration delivered at 5 Hz frequency. When the stimulus pulse duration was increased to 1 ms (5 Hz frequency) or the stimulation frequency to 50 Hz (0.04-ms duration), inhibition of [3H]acetylcholine release became evident after blockade of L-type VDCC, with nifedipine (1 microM), and/or depletion of thapsigargin-sensitive internal stores. The inhibitory effect of thapsigargin (2 microM) was still observed in Ca(2+)-free medium. Neither omega-conotoxin GVIA (1 microM) nor omega-conotoxin MVIIC (150 nM) modified neurotransmitter release. The results suggest that, depending on the stimulus paradigm, both internal (thapsigargin-sensitive) and external (either P- or L-type channels) Ca(2+) pools can be mobilized to promote acetylcholine release from motor nerve terminals. PMID- 11040343 TI - Neurobehavioral activity in mice of N-vanillyl-arachidonyl-amide. AB - We studied the cannabimimetic properties of N-vanillyl-arachidonoyl-amide (arvanil), a potential agonist of cannabinoid CB(1) and capsaicin VR(1) receptors, and an inhibitor of the facilitated transport of the endocannabinoid anandamide. Arvanil and anandamide exhibited similar affinities for the cannabinoid CB(1) receptor, but arvanil was less efficacious in inducing cannabinoid CB(1) receptor-mediated GTPgammaS binding. The K(i) of arvanil for the vanilloid VR(1) receptor was 0.28 microM. Administered i.v. to mice, arvanil was 100 times more potent than anandamide in producing hypothermia, analgesia, catalepsy and inhibiting spontaneous activity. These effects were not attenuated by the cannabinoid CB(1) receptor antagonist N-(piperidin-1-yl)-5-(4-chloro phenyl)-1-(2, 4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide.HCl (SR141716A). Arvanil (i.t. administration) induced analgesia in the tail-flick test that was not blocked by either SR141716A or the vanilloid VR(1) antagonist capsazepine. Conversely, capsaicin was less potent as an analgesic (ED(50) 180 ng/mouse, i.t.) and its effects attenuated by capsazepine. The analgesic effect of anandamide (i.t.) was also unaffected by SR141716A but was 750-fold less potent (ED(50) 20.5 microg/mouse) than capsaicin. These data indicate that the neurobehavioral effects exerted by arvanil are not due to activation of cannabinoid CB(1) or vanilloid VR(1) receptors. PMID- 11040344 TI - Effects of naltrexone on the accumulation of L-3, 4-dihydroxyphenylalanine and 5 hydroxy-L-tryptophan and on the firing rate induced by acute ethanol administration. AB - In order to characterize the effects of naltrexone, a mu-opioid receptor antagonist, on acute ethanol-induced functional modification of dopaminergic neurons in the nigrastriatal and mesolimbic dopamine systems, the accumulation of L-3, 4-dihydroxyphenylalanine (L-DOPA) in the cerebral cortex, dorsal striatum and nucleus accumbens and of 5-hydroxy-L-tryptophan (5-HTP) in the hippocampus was measured in normal rats using the mu-hydroxybenzylhydrazine dihydrochloride (NSD-1015) enzymatic inhibition method. In addition, the firing rates of dopaminergic neurons were recorded in the substantia nigra and ventral tegmental area. Naltrexone resulted in a decrease in the dopaminergic neuronal firing rates activated by ethanol and eventually in a reduction of the dopamine synthesis induced by ethanol in the dorsal striatum and nucleus accumbens, but not in the cerebral cortex. Mesolimbic dopamine neurons were slightly more sensitive to ethanol and naltrexone than were nigrostriatal dopamine neurons. The widespread inhibitory action of naltrexone also decreased the ethanol-induced stimulation of hippocampal serotonin synthesis. PMID- 11040345 TI - Opposite effects of T- and L-type Ca(2+) channels blockers in generalized absence epilepsy. AB - The role of the T-type Ca(2+) channel blocker, ethosuximide, the L-type Ca(2+) channel blocker, nimodipine and L-type Ca(2+) channel opener, BAY K8644 (1,4 Dihydro-2, 6-dimethyl-5-nitro-4-[trifluoromethyl)-phenyl]-3-pyridine carboxylic acid methyl ester), was investigated on spike-wave discharges in WAG/Rij rats. This strain is considered as a genetic model for generalized absence epilepsy. A dose-dependent decrease in the number of spike-wave discharges was found after i.c.v. ethosuximide, an increase after i.p. nimodipine and a decrease after i.c.v. BAY K8644. BAY K8644 was also able to antagonise the effects of nimodipine. Preliminary data were obtained with two conotoxins, MVIIC and GVIA, which block P/Q-type and N-type Ca(2+) channels, respectively. Only after i.c.v. administration of omega-conotoxin GVIA were the number and duration of spike-wave discharges reduced, but animals showed knock-out lying. The latter suggests behavioural or toxic effects and that the decrease in spike-wave activity cannot unequivocally be attributed to blockade of N-type Ca(2+) channels. It can be concluded that T- and L-type Ca(2+) channel blockers show opposite effects on spike-wave discharges. Furthermore, these effects are difficult to explain in terms of a model for spindle burst activity in thalamic relay cells proposed by McCormick and Bal [Sleep and arousal: thalamocortical mechanisms. PMID- 11040346 TI - Role of adenosine in the spinal antinociceptive and morphine modulatory actions of neuropeptide FF analogs. AB - The neuropeptide FF (Phe-Leu-Phe-Gln-Pro-Gln-Arg-Phe-NH(2)) and its synthetic analogs bind to specific receptors in the spinal cord to produce antinociceptive effects that are partially attenuated by opioid antagonists, and at sub-effective doses neuropeptide FF receptor agonists augment spinal opioid antinociception. Since adenosine plays an intermediary role in the production of spinal opioid antinociception, this study investigated whether this purine has a similar role in the expression of spinal effects produced by neuropeptide FF receptor agonists. In rats bearing indwelling spinal catheters, injection of adenosine receptor agonists, N6-cyclohexyladenosine (CHA, 1.72 nmol) and N ethylcarboxiamidoadenosine (NECA, 1.95 nmol), as well as morphine (13.2 nmol) elicited antinociception in the tail-flick and paw-pressure tests. Pretreatment with intrathecal 8-phenyltheophylline (5.9 and 11.7 nmol), an adenosine receptor antagonist, blocked the effect of all three agents without influencing baseline responses. Administration of two synthetic neuropeptide FF (NPFF) analogs, [D Tyr(1),(NMe)Phe(3)]NPFF (1DMe, 0. 86 nmol) and [D-Tyr(1),D-leu(2),D-Phe(3)]NPFF (3D, 8.6 nmol) produced sustained thermal and mechanical antinociception. Pretreatment with doses of intrathecal 8-phenyltheophylline (5.9, 11. 7 and 23.5 nmol), producing adenosine receptor blockade, significantly inhibited the antinociceptive effects of 1DMe or 3D. Injection of a sub-antinociceptive dose of 1DMe (0.009 nmol) significantly augmented the antinociceptive effect of intrathecal morphine (13.2 nmol) in the tail-flick and paw-pressure tests. Intrathecal 8-phenyltheophylline (11.7 nmol) reduced the effect of this combination. Administration of low dose of 1DMe (0.009 nmol) or 3D (0.009 nmol) very markedly potentiated the antinociceptive actions of the adenosine receptor agonist, N6-cyclohexyladenosine (0. 43, 0.86 and 1.72 nmol) in the tail-flick and paw-pressure tests 50 min after injection. The results suggest that the antinociceptive and morphine modulatory effects resulting from activation of spinal NPFF receptors could be due to an increase in the actions or availability of adenosine. PMID- 11040347 TI - Effects of the prototypical mGlu(5) receptor antagonist 2-methyl-6 (phenylethynyl)-pyridine on rotarod, locomotor activity and rotational responses in unilateral 6-OHDA-lesioned rats. AB - In the present study, we evaluated the effect of the prototypical metabotropic glutamate receptor 5 (mGlu(5)) antagonist 2-methyl-6-(phenylethynyl)-pyridine (MPEP) on motor behaviour in rats using the accelerating rotarod, spontaneous locomotor activity and the 6-hydroxy-dopamine (6-OHDA) lesion model to assess its treatment potential for Parkinson's disease. The data indicate that MPEP at doses between 7.5 and 300 mg/kg, p.o. did not disrupt endurance performance on the accelerating rotarod (4-40 rpm in 300 s) which indicates that MPEP has a relatively high safety margin. However, while ineffective at doses of 3.75, 7.5 and 15 mg/kg (p.o.) MPEP inhibited spontaneous locomotor activity at doses of 30 and 100 mg/kg (p.o.). In the 6-OHDA rat rotation model, at doses of 7.5, 15 and 30 mg/kg (p.o.), MPEP induced a dose-dependent ipsilateral rotational response that reached statistical significance at the highest dose tested. This effect was relatively small but consistent. In combination with direct or indirect dopamine agonists, i.e. apomorphine (0.25 mg/kg, s.c.) and D-amphetamine (2.5 mg/kg, i.p.), MPEP (7.5, 15 or 30 mg/kg, p.o.) was found to significantly inhibit these dopamine receptor mediated rotational responses. MPEP injected at a dose of 30 mg/kg also inhibited the rotational response induced by L-DOPA (25 mg/kg, i.p.). (+)MK-801 was used in these rotation experiments as the reference compound. In view of these findings, it could be concluded that MPEP and potentially other mGlu(5) receptor antagonists are probably not appropriate drug candidates for the symptomatic treatment of Parkinson's disease. PMID- 11040348 TI - Monoamine oxidase inhibitors reduce conditioned fear stress-induced freezing behavior in rats. AB - The present study examined the acute anxiolytic effects of monoamine oxidase inhibitors on freezing behavior, a putative index of anxiety induced by conditioned fear stress. The selective serotonin 1A receptor agonist tandospirone (0.1-10 mg/kg) inhibited freezing dose dependently. The irreversible, non selective monoamine oxidase inhibitors tranylcypromine (3 and 15 mg/kg) and phenelzine (30 and 80 mg/kg) reduced freezing significantly. Clorgyline (10 mg/kg, irreversible selective monoamine oxidase A inhibitor), N-(2-aminoethyl)-5 (m-fluorophenyl)-4-thiazole carboxamide (Ro 41-1049) (30 mg/kg, reversible selective monoamine oxidase A inhibitor), selegiline (3 mg/kg, irreversible selective monoamine oxidase B inhibitor) and lazabemide (10 mg/kg, reversible selective monoamine oxidase B inhibitor) had no effect on freezing behavior. However, combined administration of clorgyline (10 mg/kg) and selegiline (3 mg/kg) reduced freezing significantly, as well as combined administration of clorgyline (10 mg/kg) and lazabemide (10 mg/kg), Ro 41-1049 (30 mg/kg) and selegiline (3 mg/kg), or Ro 41-1049 (30 mg/kg) and lazabemide (10 mg/kg). These effects of monoamine oxidase inhibitors on freezing were not due to non-specific motor effects. These results suggest that acute inhibition of both monoamine oxidase A and B reduced anxiety or fear, while inhibition of monoamine oxidase A or B alone failed to reduce anxiety or fear. PMID- 11040349 TI - Effects of ouabain on the pressor response to phenylephrine and on the sodium pump activity in diabetic rats. AB - The diabetes mellitus insulin-dependent is usually associated with cardiovascular disorders and with changes in the activity of the Na(+),K(+)-ATPase. The effects of ouabain, a Na(+),K(+)-ATPase inhibitor, on the pressor response of 7-day streptozotocin-induced diabetes were investigated in anesthetized rats and on the vascular reactivity of the perfused rat tail vascular bed. Diabetes was characterized by hyperglycemia (86+/-7.8 vs. 471+/-18.5 mg/dl) without changes in arterial blood pressure. Blood pressure increased after the treatment with 18 microg/kg ouabain in controls but not in diabetic rats; acute hyperglycemia, in non-treated rats, did not change these effects. Control tail vascular beds showed increased maximal response to phenylephrine after treatment with 10 nM ouabain for 1 h; this response was abolished in streptozotocin-treated rats. These rats showed an increased sensitivity to phenylephrine without changing the maximal vasoconstrictor response when compared to control rats. The relaxation induced by acetylcholine was reduced in diabetic rats. The functional activity of the Na(+),K(+)-ATPase was inhibited in vascular beds from diabetic rats, when compared to control rats, and the inhibition of the Na(+),K(+)-ATPase with 10 nM ouabain was not effective in these rats. Results suggested that in 7-day diabetic rats, the increase of arterial blood pressure or the sensitization of the vascular bed produced by ouabain is lost as a consequence of the reduction of the functional activity of the Na(+), K(+)-ATPase probably as a result of insulin lack and a deficient endothelial nitric oxide activity. PMID- 11040350 TI - Co-incubation of native and oxidized low-density lipoproteins: potentiation of relaxation impairment. AB - The influence of native low-density lipoprotein (LDL) on the inhibition of endothelium-dependent relaxation previously induced by oxidized LDL was investigated with intact rabbit aortic rings. We also tried to assess oxysterol involvement in the native lipoprotein effects. Lipoprotein fractions (1 mg protein/ml) were tested for their ability to inhibit the vasorelaxation induced by acetylcholine in aorta rings previously precontracted by noradrenaline vs. that in control strips in Krebs buffer. Co-incubation of oxidized and native LDL reinforced the oxidized LDL-induced inhibition, compared to the impairment evoked by oxidized LDL alone (E(max)=43.3+/-6.7% and 61. 4+/-5.4%, respectively; P<0.05). Finally, smaller amounts of 7-oxy-cholesterols were recovered in organ baths after co-incubation of native and oxidized LDL than after incubation of oxidized LDL alone. Conversely, more oxy-cholesterols were found in the strip vessels under the same conditions (% of oxysterol incorporation: 0. 05158 vs. 0.10199, r=0.703). Together these results suggest that the strengthening of oxidized LDL-induced inhibition by native LDL is dependent on an oxysterol effect on arterial wall cells. Mechanisms involved in this phenomenon remain to be investigated. PMID- 11040351 TI - Effect of cortisol on fetal ovine vascular angiotensin II receptors and contractility. AB - The renin angiotensin system is important in the regulation of fetal blood pressure. This study investigated the expression of angiotensin AT(1) and AT(2) receptors in the ovine fetal heart, aorta and umbilical artery, and how these receptors are affected by cortisol. Cortisol infusion into the fetus has previously been shown to cause an increase in fetal blood pressure. We hypothesised that this effect of cortisol is mediated by upregulation of the angiotensin AT(1) receptor. Binding studies performed on tissues with intact endothelium demonstrated both receptor subtypes in the fetal aorta and right ventricle, although the latter contained mainly angiotensin AT(2) receptors. In contrast, only angiotensin AT(1) receptors were found in the umbilical artery. Cortisol infusion into fetuses (3 mg/day for 3-5 days) caused a physiological increase in plasma cortisol levels to 29+/-4 nM. This was associated with an increase in systolic pressure (57.8+/-1.7 vs. 52.2+/-1.5 mm Hg, P<0.05), but cortisol had no effect on the density or affinity of angiotensin receptors, nor on the in vitro contractile responses of carotid and umbilical arterial rings to 5-microM angiotensin II. In conclusion, this study has demonstrated differential expression of angiotensin AT(1) and AT(2) receptors in the different regions of the ovine fetal cardiovascular system and that the angiotensin AT(1) receptor is functional. The lack of any effect of low doses of cortisol on these receptors and on the contractility of isolated fetal vessels to angiotensin II suggests cortisol acts by other mechanisms to raise fetal arterial pressure. PMID- 11040352 TI - Effects of sparfloxacin, grepafloxacin, moxifloxacin, and ciprofloxacin on cardiac action potential duration. AB - Fluoroquinolone antibiotics have been associated with QT prolongation following administration to humans. This study compares the effects of four fluoroquinolones, sparfloxacin, grepafloxacin, moxifloxacin and ciprofloxacin on action potential duration recorded from canine isolated cardiac Purkinje fibres. Left and right ventricular Purkinje fibres were isolated from canine hearts and continuously superfused with physiological salt solution. Action potential duration at 90% repolarization was recorded via intracellular microelectrodes. Sparfloxacin, grepafloxacin, moxifloxacin and ciprofloxacin prolonged action potential duration in a concentration dependent manner. Mean concentrations causing a 15% prolongation of action potential duration recorded at a stimulation frequency of 1 Hz were: sparfloxacin 4.2+/-0.7 microg/ml; grepafloxacin 9.3+/-0.9 microg/ml; moxifloxacin 9.9+/-1.6 microg/ml and ciprofloxacin 72.8+/-26.4 microg/ml. Prolongation was inverse frequency dependent with larger increases in action potential duration occurring when the stimulation frequency was reduced to 0.5 Hz. These results indicate that effects on action potential duration vary within this class of compound. Rank order of potency was sparfloxacin > grepafloxacin = moxifloxacin > ciprofloxacin. PMID- 11040353 TI - Expression and function of P-glycoprotein in rats with glycerol-induced acute renal failure. AB - The effect of glycerol-induced acute renal failure on P-glycoprotein expression and function was evaluated in rats. The in vivo function of P-glycoprotein was evaluated by measuring renal secretory and biliary clearance and brain distribution of rhodamine 123 (Rho-123), a P-glycoprotein substrate, under a steady-state plasma concentration. In acute renal failure rats, the P glycoprotein level increased 2.5-fold in the kidney, but not in the liver and brain. In contrast, P-glycoprotein function in these tissues was suppressed. Interestingly, not only the renal but also the biliary clearance of Rho-123 was correlated with the glomerular filtration rate. In Caco-2 cells, plasma from renal failure rats exhibited a greater inhibitory effect on P-glycoprotein mediated transport of Rho-123 than did plasma from control rats. In conclusion, P glycoprotein function was systemically suppressed in acute renal failure, even though the level of P-glycoprotein remained unchanged or rather increased. This may be due to the accumulation of some endogenous P-glycoprotein substrates/modulators in the plasma in disease states. PMID- 11040354 TI - Nonsteroidal anti-inflammatory drug-induced acute gastric injury in Helicobacter pylori gastritis in Mongolian gerbils. AB - We examined the acute ulcerogenic effects of indomethacin and N-(2, cyclohexyloxy 4-nitrophenyl)methane sulfonamide (NS-398) on the gastric mucosa in Helicobacter pylori-infected Mongolian gerbils. H. pylori infection for 4 and 12 weeks caused moderate and severe gastritis, respectively, with cyclooxygenase-2 expression and an increase in prostaglandin E(2) production. In normal animals, gastric injury was caused by indomethacin, but not by NS-398. At 4 weeks infection, gastric lesions were synergistically aggravated by indomethacin, and NS-398 at high doses. However, at 12 weeks, the synergistic effects of indomethacin and NS-398 with H. pylori were not observed. Indomethacin and NS-398 at high doses inhibited prostaglandin E(2) production in both normal and the infected mucosa. NS-398 at low dose reduced only the H. pylori-increased prostaglandin production. These results suggest that nonsteroidal anti-inflammatory drugs (NSAIDs) synergistically aggravate gastric lesions in moderate H. pylori gastritis, but not in severe gastritis. Cyclooxygenase-2 inhibition only might not induce acute gastric injury in H. pylori gastritis. PMID- 11040355 TI - Effects of wortmannin on bronchoconstrictor responses to adenosine in actively sensitised Brown Norway rats. AB - The bronchoconstrictor response to adenosine in the actively sensitised Brown Norway rat is markedly augmented following low level allergen (ovalbumin) challenge. The response reflects activation of the A(2B) receptor subtype and is mediated by 5-hydroxytryptamine (5-HT) released as a consequence of mast cell activation. We describe here the effects of wortmannin, a potent inhibitor of phosphatidylinositol-3-kinase and mast cell exocytosis, on the response to adenosine. Bronchoconstrictor responses to adenosine elicited 3 h following ovalbumin challenge were markedly and dose-dependently reduced by wortmannin given intratracheally (i. t.), 1 h prior to or 2 h post-allergen challenge. Responses to methacholine, which activates bronchial smooth muscle directly, and 5-HT were also reduced following wortmannin but to a lesser extent than those to adenosine. Bronchoconstrictor responses to adenosine 3 h post-challenge with vehicle were also markedly reduced by wortmannin given intratracheally (i.t.), 1 h prior to the "sham" challenge. Plasma histamine and 5-HT levels increased in response to adenosine given 3 h following ovalbumin challenge. The increases were suppressed by wortmannin given i.t., 2 h post-ovalbumin challenge. A reduction in the sensitivity of the airways to 5-HT explains in part the reduced bronchoconstrictor response to adenosine induced by wortmannin. A direct action to suppress 5-HT release from airway mast cells induced by adenosine also contributes to the reduction in the response. Inhibition of phosphatidylinositol 3-kinase is the presumed mechanistic basis for the observed effects. PMID- 11040356 TI - Wogonin inhibits inducible prostaglandin E(2) production in macrophages. AB - Effects of 5,7-dihydroxy-8-methoxyflavone (wogonin) on cyclooxygenase-2 (COX-2) mediated prostaglandin E(2) production in macrophages were investigated. Stimulation with lipopolysaccharide (LPS; 1 microg/ml) greatly increased prostaglandin E(2) production in RAW 264.7 murine macrophages. The stimulated prostaglandin E(2) production was abolished in the presence of indomethacin (1 microM) or cycloheximide (2 microM), suggesting that the increased production of prostaglandin E(2) by LPS reflects the inducible synthesis of prostaglandin E(2) by COX-2. Wogonin (0.1-50 microM) concentration-dependently inhibited inducible prostaglandin E(2) production. Wogonin at concentrations as low as 0.5 microM directly attenuated enzymatic activity of COX-2. The protein expression of COX-2 was depressed by wogonin at concentrations of 10 microM and more. These results suggest that wogonin decreases inducible prostaglandin E(2) production in macrophages by inhibiting both COX-2 activity and COX-2 expression. The former action requires much lower doses of wogonin. These wogonin actions may explain, in part, its anti-inflammatory action. PMID- 11040357 TI - Energy-dependent UV light-induced disruption of (-)sulpiride antagonism of dopamine. AB - The dopamine D2 receptor antagonist sulpiride decreases the spontaneous locomotor activity of Planaria in an enantiomeric-selective and dose-dependent manner. We now report that (-)sulpiride (0.1 microM)-induced decrease of planarian locomotor activity is significantly (P<0.05) attenuated by low-energy (366 nm) ultraviolet (UV) light and to a greater extent by high-energy (254 nm) UV light. The phenomenon offers a novel approach for studying dopamine D2 receptor transduction processes in a simple in vivo model. PMID- 11040359 TI - The 100th volume of the journal of neuroscience methods PMID- 11040358 TI - Suppression of feeding-evoked dopamine release in the rat nucleus accumbens by the blockade of P(2) purinoceptors. AB - In order to investigate whether endogenous adenosine 5'-triphosphate (ATP) is involved in the regulation of feeding, the influence of the P(2) receptor antagonist pyridoxalphospate-6-azophenyl-2', 4'-disulphonic acid (PPADS) infused into the rat nucleus accumbens on 18-h food-deprived feeding was tested. PPADS suppressed the feeding-induced dopamine release and reduced the amount of food consumed as well as the time of feeding. These results indicate that activation of P(2) purinoceptors by endogenous ATP facilitates feeding behaviour and contributes to the feeding-associated dopamine release. PMID- 11040360 TI - A system for the study of visuomotor coordination during reaching for moving targets. AB - Prehensile behavior is a popular task in current research on human motor control. Most studies on reaching used stationary target objects and, therefore, most models do not address the challenges the motor system must respond to when reaching for moving objects. The machines used in earlier studies to produce object motion offered a limited range of trajectories and restricted control over various movement parameters. We have developed a device that allows a great variety of object trajectories along a flat-table surface and gives the experimenter full control over all movement parameters. A linear positioning system is used to move a sled beneath the table surface. Magnetic coupling transfers the sled's movement to the target object on the tabletop. This arrangement allows fast movements of the object (up to 5 m/s) and at the same time protects subjects from any harm due to the moving parts. The system is connected to LC shutter glasses, a 3-D movement registration device, and a switch that detects the onset of hand motion. This allows the selective withdrawal of vision during the reaching task or the introduction of changes in the object motion depending on the subject's reactions. PMID- 11040361 TI - Pressure polishing: a method for re-shaping patch pipettes during fire polishing. AB - The resolution of patch-clamp recordings is limited by the geometrical and electrical properties of patch pipettes. The ideal whole-cell patch pipette has a blunt, cone-shaped tip and a low resistance. The best glasses for making patch pipettes are low noise, low capacitance glasses such as borosilicate and aluminasilicate glasses. Regrettably, nearly all borosilicate glasses form pipettes with sharp, cone-shaped tips and relatively high resistance. It is possible, however, to reshape the tip during fire polishing by pressurizing the pipette lumen during fire polishing, a technique we call 'pressure polishing.' We find that this technique works with pipettes made from virtually any type of glass, including thick-walled aluminasilicate glass. We routinely use this technique to make pipettes suitable for whole-cell patch-clamp recording of tiny neurons (1-3 microm in diameter). Our pipettes are made from thick-walled, borosilicate glass and have submicron tip openings and resistances <10 MOmega. Similar pipettes could be used to record from subcellular neuronal structures such as axons, dendrites and dendritic spines. Pressure polishing should also be useful in patch-clamp applications that benefit from using pipettes with blunt tips, such as perforated-patch whole-cell recordings, low-noise single channel recordings and experiments that require internal perfusion of the pipette. PMID- 11040362 TI - Annealing of the Co/Hf bilayer on single Si, polycrystalline Si and SiO(2). AB - Silicidation of the Co/refractory metal/Si system in which the refractory metal is used as an epitaxy promoter for CoSi(2) has recently received much attention. Hf is one of the candidates for the epitaxy promoter of cobalt silicide like Ti. In this paper, we investigated the layer structures of the Co/Hf bilayer on various substrates like single (100)Si, polycrystalline Si and SiO(2) after rapid thermal annealing. Epitaxy of CoSi(2) was obtained on (100)Si by annealing Co/Hf/(100)Si. Co-Hf compounds seem to play an important role of barriers against the reaction between Co and Si during silicidation of Co/Hf/(100)Si. The existence of Co-Hf compounds is helpful in the formation of epitaxial CoSi(2) since they retard the diffusion of cobalt and silicon atoms. The transition temperatures of cobalt silicides in the Co/Hf/poly-Si system were found to be lower than those in the Co/Hf/(100)Si system. The reaction between the metal and the spacer SiO(2) during silicidation is a matter of concern since any conducting residue of this reaction could degrade oxide integrity or produce bridging. In the Co/Hf/SiO(2) system Hf oxides formed as a result of the reaction between Hf and SiO(2) but a conducting material like HfSi(2) was not found to form after annealing. Considering the temperature at which epitaxial CoSi(2) forms along with the one from which the SiO(2) substrate starts collapsing we may conclude that the optimum silicidation annealing temperature for Co/Hf/Si is 600 degrees C. PMID- 11040363 TI - A simple procedure for quantification of neurite outgrowth based on stereological principles. AB - The molecular mechanisms controlling formation and remodelling of neuronal extensions are of considerable interest for the understanding of neuronal development and plasticity. Determination of neurite outgrowth in cell culture is a widely used approach to investigate these phenomena. This is generally done by a time consuming tracing of individual neurites and their branches. We have used stereological principles to determine the length of neurites. The total neuritic length per cell was estimated by counting the number of intersections between neurites and test lines of an unbiased counting frame superimposed on images of cell cultures obtained by conventional computer-assisted microscopy. The absolute length, L, of neurites per cell was subsequently estimated from the number of neurite intersections, I, per cell by means of the equation L=(pid/2)I describing the relationship between the number of neurite intersections and the vertical distance, d, between the test lines used. When measuring neurite outgrowth from PC12 cells and primary hippocampal neurons, data obtained by counting neuritic intersections correlated statistically significantly with data obtained using a conventional tracing technique. However, information was acquired more efficiently using the stereological approach. Thus, using the described set-up, the stereological procedure was approximately five times less time consuming than the conventional method based on neurite tracing. The study shows that stereological estimation of neuritic length provides a precise and efficient method for the study of neurite outgrowth in cultures of primary neurons and cell lines. PMID- 11040364 TI - The use of a sodium tungstate developer markedly improves the electron microscopic localization of zinc by the Timm method. AB - The Timm's sulfide-silver method is frequently used for the demonstration of the mossy fiber bundle or sprouted mossy fibers in the normal or epileptic hippocampal dentate gyrus. Under the light microscope the results are excellent, but the ultrastructure is considerably impaired and the silver grains produced are too large as compared to the sizes of intra-synaptic structures. The present study was meant to test a series of physical developers containing, instead of gum arabic, sodium tungstate as protective colloid. One of them left the ultrastructure fairly intact and produced small, round silver grains, making it possible to precisely locate zinc in mossy terminals. With this method, it could be demonstrated that zinc is contained inside synaptic vesicles in the resting axon terminals of granule cells. As a consequence of prolonged sodium sulfide perfusion, zinc is released from synaptic vesicles and enters the synaptic cleft. PMID- 11040365 TI - Cross-correlation measures of unresolved multi-neuron recordings. AB - An increasing number of laboratories are studying population properties of the nervous system using data where the spike activity of more than one neuron is recorded on each electrode and where, accidentally or deliberately, these activities are not resolved into single unit spike trains. We have previously examined the consequences for measurement of cross-correlation between two such electrodes in the limited case where all individual distant (between electrode) correlations are the same and all individual close (on a single electrode) correlations are the same [Bedenbaugh, P.H., and Gerstein, G.L. (1997). Multiunit normalized cross correlation differs from the average single-unit normalized correlation. Neural Computation 9, 1265-1275]. Here, we lift these unrealistic restrictions to allow all values of individual correlation, and examine explicitly the cases of two or three unresolved neurons on each electrode. In these situations, the cross-correlation coefficient measured between the electrodes is a linear sum of the distant correlations, divided by a non-linear function of the close correlations. We then examine in detail the case of a single direct distant correlation and take account of all relevant indirect correlations. The measured interelectrode correlation shows a reduction of this actual distant correlation by a non-linear function of the close correlations on each electrode over most of their possible values. Finally, we examine the consequences of poor waveform sorting for correlation measures; here a supposedly isolated spike train is contaminated by some fraction of the activity of another train, a situation that unfortunately is all too common in experiments. All these distortions become far more serious in the more realistic situation of dynamic firing rates and correlations. This paper is intended as a cautionary note for those who want to draw inferences about neuronal organization and/or coding or representation by using cross-correlation analysis of unresolved recordings. PMID- 11040366 TI - Determination of the spatial distribution of major elements in the rat brain with X-ray fluorescence analysis. AB - An energy dispersive X-ray fluorescence analysis was applied for determining the spatial (two-dimensional) distribution of elemental concentrations in rat brain sections. Freeze-dried brain sections prepared from normal and ischemic rats with middle cerebral artery occlusion were scanned with a collimated X-ray beam (0.18 mm in diameter, 50-kV acceleration voltage). The fluorescent Kalpha X-rays of P, S, Cl, and K were detectable, so that the two-dimensional distribution of fluorescent X-ray intensities could be determined for these elements. Furthermore, quantitative determination was possible for P and K by using the fundamental parameter technique. However, the accurate determination of Na and Ca was difficult, because of the low energy of Kalpha X-ray of Na, and the interference of K-Kbeta with Ca-Kalpha. The change in elemental concentrations in ischemic tissue, including the decrease in K concentration and increase in Cl concentration, was demonstrated by this method as a two-dimensional contour map. Since it is possible to obtain a pictorial representation of the elemental concentration in tissue sections, this method may be useful to evaluate the ionic changes in injured brain tissue in relation to histological or autoradiographical observations. PMID- 11040367 TI - Combined use of the green and yellow fluorescent proteins and fluorescence activated cell sorting to select populations of transiently transfected PC12 cells. AB - One of the more time-consuming procedures in the study of exogenously expressed proteins in cell lines is the selection of individual transfected clones. In recent years, green fluorescent protein variants with excitation/emission spectra matching the typical flow cytometer configurations have been generated and are in common use. We employed PC12 cells transfected with vectors encoding fluorescent proteins and a fluorescence selection procedure using a fluorescence-activated cell-sorter. In order to select the optimal co-electroporation and sorting conditions, we used the simultaneous detection of two variants of the green fluorescent protein, that possess separable emission peaks when excited at 488 nm. Using these variants and the adequate combination of band-pass filters, we were able to analyze and establish the conditions for identifying and sorting cells transfected with enhanced green fluorescent protein, that simultaneously express another plasmid of interest. Using this procedure, the cells sorted that express both plasmids exceeded 90%. The whole procedure did not alter the physiological responsiveness of the transfected cells to growth factors, and has been successfully applied to the constitutive activation of the mitogen-activated protein kinase pathway, resulting in the spontaneous differentiation of PC12 cells. Also, this procedure has been used with other set of expression vectors encoding proteins that protect PC12 cells from apoptosis caused by different stimuli. The method that we present here provides an easy and fast procedure to obtain a high proportion of positively transfected populations of PC12 cells. PMID- 11040368 TI - A spinal cord slice preparation for analyzing synaptic responses to stimulation of pelvic and pudendal nerves in mature rats. AB - The dorsal commissural nucleus (DCN) in the lumbosacral spinal cord (L6-S1) receives primary afferent fibers from both pelvic and pudendal nerves in rats. However, the physiological and pharmacological properties of synaptic responses of the DCN neurons to stimulation of those nerves remain unclear. We have developed a longitudinal spinal cord (L6-S1) slice preparation from mature rats that retained both nerves attached. Blind whole-cell recordings were made from the DCN neurons in this preparation. In most neurons, mono- and/or poly-synaptic fast excitatory postsynaptic potentials (EPSPs) were evoked by electrical stimulation of either the pelvic or pudendal nerve. These EPSPs were mediated by activation of Abeta/Adelta and/or C fibers (conduction velocities, 0.5-17.3 m/s), and were abolished by CNQX. Fast EPSPs elicited by either pelvic or pudendal nerve stimulation were occasionally accompanied by bicuculline- and strychnine sensitive IPSPs. In one-third of the neurons tested, mono- and/or poly-synaptic EPSPs were elicited by the stimulation of both the pelvic and pudendal nerves, indicating convergence of the visceral and somatic primary afferent inputs from the pelvic region onto the DCN neurons. The preparation is applicable to study the mechanism of the integration of the visceral and somatic inputs in the spinal cord. PMID- 11040369 TI - Simple methods for quantifying the spatial distribution of different categories of motoneuronal nerve endings, using measurements of muscle regionalization. AB - For skeletal muscles, a well-known match exists between the properties of motoneurones and those of their muscle fibres. Hence, the intramuscular distribution of different kinds of motoneuronal nerve endings (e.g. 'slow' versus 'fast') can be mapped by determining the distribution of the corresponding types of muscle fibre. As a background for further studies of motoneuronal plasticity, we needed precise measures of such distributions. Simple quantitative methods were developed for defining the position and extent of sub-populations of cells within a structure (e.g. the regional distribution of slow versus fast muscle fibres within a muscle cross-section): (a) The 'mass vector method' defined the relative position of the target cell cloud. A line was drawn between the calculated centre of mass for the target cells and that for the whole structure. The direction (a1) and length (a2) of this line gave a measure of the direction and degree of target cell eccentricity within the structure. (b) The 'sector method' delineated the region containing the target fibres. A circle around the centre of mass for the target fibres was subdivided into a number of equal sectors (standard setting: 20). The most remote point was found within each sector and a line joining these points defined the region of the target fibres. When applied to the 'slow' type I fibres of cross-sections from rat hindlimb muscles, the regional area estimates obtained by the sector method were highly correlated with, but approximately 10% lower than those achieved by the well established 'convex hull' method. Highly significant inter-muscular differences were observed for each one of the three new parameters described in this paper (a1, a2, b). PMID- 11040370 TI - Imprinting as a rapid technique for assessing the morphology of the central nervous system by immunofluorescence. AB - This paper describes a technique that has been developed to assess the in vivo morphology of central nervous system (CNS) tissue by immunofluorescence. This technique permits the study of tissue that is mainly just a monolayer of cells. Unlike routine cryosections that are much thicker (10-15 microm), imprinting does not section the cells, but can result in the detachment of whole cells onto a glass surface for subsequent staining. The imprinting technique is simple and rapid and does not require prior fixation or embedding of the tissue. It has been used to evaluate antigens expressed at the cell surface, in myelin and in the cytoskeleton in the studies of normal and myelin mutant mice. Using the imprinting/immunofluorescence technique one can now assay the genotype of mouse strains that differ in their expression of cell surface antigens within 2 h. PMID- 11040371 TI - The design and application of three speaker-based stimulating devices for cutaneous stimulation in anesthetized and awake animals. AB - Those wishing to study neuronal plasticity in sensory systems are confronted by the need to deliver equivalent stimuli to the organism at time intervals separated by hours, days or months. This problem is particularly acute in the somatosensory system where delivering an equivalent stimulus generally requires a second physical contact with the same point on a geometrically complex surface. This requirement is difficult to fulfill. We have designed two stimulators that avoid or minimize the importance of this requirement by obviating the need for the stimulator to be at a fixed distance from the skin. As well, we have redesigned a system for whisker stimulation originally proposed by Simons. The first stimulator is appropriate for experiments in anesthetized animals; the surface to be stimulated is immersed in water warmed to body temperature and the tactile stimulus is generated as an hydraulic pulse. The second uses a high velocity pulse of air shaped so that it can be transmitted significant distances without attenuation. The redesign of the Simons' vibrissa stimulator provides larger amplitude displacements and lower controlling voltages more readily generated by equipment normally found in laboratories. We also described the design of a chamber for restricting the awake rat during chronic study and the electrodes used for recording and for delivery of drugs in awake animals held in such a chamber. PMID- 11040372 TI - An axotomy model for the induction of death of rat and mouse corticospinal neurons in vivo. AB - To study trophic dependencies of rat and mouse corticospinal neurons (CSN), we established a lesion model for the induction of death of analogous populations of CSN in these rodent species. Before lesion, CSN were retrogradely labeled with Fast Blue (FB). A stereotaxic cut lesion through the entire internal capsule (ICL) was used to axotomize CSN. The extent of axotomy was determined by application of a control tracer. In both species, FB-labeled CSN were localized in three major areas: (1) the sensory motor cortex; (2) the supplementary motor and medial prefrontal cortex; and (3) the somatosensory cortex. ICL does not lead to complete axotomy of CSN of the rat and mouse somatosensory cortex. In rats, ICL results in complete axotomy of CSN of the sensory motor cortex and incomplete axotomy of the caudal portion of the supplementary motor and medial prefrontal cortex. In mice, the area of axotomized CSN extends significantly further frontally. In both species, axotomy-induced death of CSN is observed in the center of the sensory motor cortex. This lesion model is useful for investigations on the response of CSN of the sensory motor cortex to lesion and therapeutic drugs. PMID- 11040373 TI - Reduced long-term potentiation in hippocampal slices prepared using sucrose-based artificial cerebrospinal fluid. AB - Sucrose-based artificial cerebrospinal fluid (aCSF) is sometimes used to prepare brain slices for in vitro electrophysiological experiments. This study compared the effect of preparing brain slices using chilled sucrose-based aCSF versus the conventional method using chilled aCSF on hippocampal synaptic plasticity. Brain slices from each treatment group were transferred to normal aCSF before electrophysiological recordings were made. The stimulus-response relationship of field excitatory postsynaptic potentials (fEPSPs) in the CA1 region was indistinguishable between the two treatment groups. However, the amount of LTP induced by either a θ-burst (four stimuli at 100 Hz repeated ten times at 200 ms intervals) or tetanic stimulation (100 Hz for 1 s) was significantly reduced in slices that had been prepared using sucrose-based aCSF. This was associated with reduced facilitation of the fEPSPs during the high frequency stimulus, reduced post-tetanic potentiation and short-term potentiation. In sucrose-cut slices the fEPSPs were slightly shorter in duration (29%, P<0.01), and during paired-pulse stimulation the broadening of the second fEPSP was enhanced. The LTP deficit in sucrose-cut slices was reversed by blocking GABA(A) receptor function with picrotoxin. These data suggest that the use of sucrose based aCSF better preserves GABA-mediated synaptic transmission, which limits the induction of LTP in hippocampal brain slices. PMID- 11040374 TI - Rapid genotyping of newborn gene mutant mice. AB - One important aspect of utilizing transgenic mice is the need to genotype them in order to distinguish mice that carry a disrupted gene or a transgene from mice that do not. Current methods for genotyping include isolation of genomic DNA from tail biopsies followed by PCR amplification. Particularly, both digestion of tail tissue using proteinase K as well as resuspension of purified DNA are time consuming and were usually carried out overnight. Here, we describe a rapid and robust method for the genotyping of bdnf targeted mice which allows us to determine the genotype of newborn mice at the day of birth within 6 h. After a freezing-thawing step tail tissue is digested in less than 2 h, and the DNA is precipitated, resuspended and ready for PCR in about 60 min. The method could be easily adapted to a variety of different mutant mice and especially should benefit neuroscientists interested in using animals with known genotype very early in postnatal development. PMID- 11040375 TI - A continuous fluorometric assay for phospholipase A(2) activity in brain cytosol. AB - Alterations in phospholipase A(2) (PLA(2)) activity have been implicated in Alzheimer disease and other neurological disorders, although brain PLA(2) activity is currently measured using lengthy, non-continuous assays. We describe herein a rapid, continuous assay in which we measured PLA(2) activity in mouse brain cytosol (CB-57). Brains were homogenized in HEPES buffer (pH 7.5) and the cytosolic fraction was prepared by centrifugation at 25000xg for 20 min, followed by centrifugation of the supernatant at 100000xg for 60 min. Cytosolic protein content was determined using the Bradford assay. Pyrene labeled phosphatidylcholine was added to 50 microg of cytosolic protein in Tris buffer (pH 8.0) containing fatty acid free-bovine serum albumin for a final assay volume of 2 ml. Assay temperature was maintained at 30+/-1 degrees C. The excitation wavelength was 345 nm and emission was measured at 377 nm. Fluorescence intensity was converted to molar concentrations using a standard curve. Under these conditions, bromoenol lactone inhibited up to 58% of the PLA(2) activity with an IC(50) of 0.5 microM. In a separate experiment, lack of appreciable alternative acylhydrolase activity was verified chromatographically. Using this method, brain PLA(2) activity can be measured in a continuous, rapid, and sensitive manner. PMID- 11040376 TI - Correction methods for three-dimensional reconstructions from confocal images: I. Tissue shrinking and axial scaling. AB - We show here, using locust wholemount ganglia as an example, that scaling artifacts in three-dimensional reconstructions from confocal microscopic images due to refractive index mismatch in the light path and tissue shrinking, can account for dramatic errors in measurements of morphometric values. Refractive index mismatch leads to considerable alteration of the axial dimension, and true dimensions must be restored by rescaling the Z-axis of the image stack. The appropriate scaling factor depends on the refractive indices of the media in the light path and the numerical aperture of the objective used and can be determined by numerical simulations, as we show here. In addition, different histochemical procedures were tested in regard to their effect on tissue dimensions. Reconstructions of scans at different stages of these protocols show that shrinking can be avoided prior to clearing when dehydrating ethanol series are carefully applied. Fixation and mismatching buffer osmolarity have no effect. We demonstrate procedures to reduce artifacts during mounting and clearing in methyl salicylate, such that only isometric shrinkage occurs, which can easily be corrected by rescaling the image dimensions. Glycerol-based clearing agents produced severe anisometric and nonlinear shrinkage and we could not find a way to overcome this. PMID- 11040377 TI - Intracellular recording of lamina X neurons in a horizontal slice preparation of rat lumbar spinal cord. AB - A horizontal slice preparation of postnatal rat lumbar spinal cord has been developed which allows correlative observations of the morphology, electrophysiology, and receptor pharmacology of lamina X neurons. These slices better maintain afferent input and somatodendritic morphology and are amenable to subsequent immunohistochemical processing. Stable intracellular recordings obtained from postnatal day 14-45 animals reveal that a number of different intrinsic membrane conductances contribute to the regulation of excitability in lamina X neurons. In addition, lamina X neurons possess inhibitory GABAergic as well as excitatory glutamate and cholecystokinin receptors. This preparation will be useful in future studies designed to characterize developmental changes in the intrinsic membrane properties, synaptic profiles and neuropeptide responsiveness of lamina X neurons in the rat. Such a characterization is important given that lamina X represents a unique sexually dimorphic region that is a convergence site for somatic and visceral afferent inputs, which includes nociceptive information. PMID- 11040378 TI - A highly sensitive and selective radioimmunoassay for the measurement of neurotensin. AB - A highly selective and sensitive radioimmunoassay (RIA) for the detection of endogenous neurotensin (NT) has been developed. We have raised a C-terminally directed antibody (CAb) that specifically binds 'biologically active' NT (NT and NT(8-13)) and that does not significantly cross-react with inactive NT metabolites or other bioactive peptides in the CNS. By reducing the volume of the assay to a low volume-RIA (30 microl), such that in vivo measurements can be made, we have increased the sensitivity (<0.3 fmol per tube), with inter- and intra-assay variations of 11.2 and 5.8%, respectively. Comparisons with similar methods of detecting NT have demonstrated that this RIA has a higher sensitivity than previously used RIA's and ELISA's. The data presented suggests that this sensitive RIA is a reliable method ideal for the detection of small quantities of biologically active NT. PMID- 11040379 TI - A microtiter trypan blue absorbance assay for the quantitative determination of excitotoxic neuronal injury in cell culture. AB - An automated method for the determination of neuronal cell death using trypan blue is described. Following various excitotoxic insults, murine mixed cortical cell cultures are stained with trypan blue (0.05%; 15 min), followed by SDS (1%) lysis. The absorbance of the dye is measured spectrophotometrically at 590 nm using a microtiter plate reader. When compared to the biochemical lactate dehydrogenase assay, no statistical difference in the calculated levels of excitotoxic neuronal cell death was noted between the assays in any given paradigm. This method is fast and reliable. It eliminates the need for cell counting, thus allowing for high volume sample analysis with a minimum of sample error. Utility of this trypan blue absorbance spectrophotometric assay is likely to extend beyond the study of excitotoxic neuronal injury and should complement existing methods for measuring neuronal viability and cytotoxicity in cell culture. PMID- 11040380 TI - A highly sensitive opto-electronic system for the measurement of movements. AB - An opto-electronic system has been developed to measure movements of insect appendages. It is made from a mirror-lens and a linear position-sensitive photodiode. The design of the mirror-lens has been exploited to axially mount a high intensity halogen light source in front of the mirror-lens. The system monitors a reflective marker which is attached to the moving object. Upon illumination by the light source the reflected light is picked up by the optical system and is focussed on the diode. The diode provides a voltage output proportional to the distribution of the light on it's surface. Since the marker is the brightest spot in the image the output of the system corresponds to the position of the marker. At a working distance of 80 cm appendage movements with amplitudes from 10 microm to 20 mm peak-peak amplitude can be recorded. The system accurately detects movements ranging from slow positional changes to 5 kHz oscillations. Currently it used to measure the stridulatory wing movements of crickets but may be applied to a variety of movement recordings. PMID- 11040381 TI - On the application, estimation and interpretation of coherence and pooled coherence. PMID- 11040382 TI - Studies on energetic compounds part 8 : thermolysis of salts of HNO(3) and HClO(4). AB - The thermolysis of various substituted ammonium salts of nitric and perchloric acids has been reviewed in the present communication. The mechanistic aspects of thermal decomposition of these salts have been discussed critically. It has been observed that the proton transfer process do play a major role during thermolysis of these salts. The plausible decomposition pathways have also been described. PMID- 11040383 TI - Incompatibilities of chemicals. AB - Chemical incompatibilities are potentially significant problems where hazardous chemicals are found. A number of chemical segregation systems have been developed which provide recommendations for the separation of incompatible chemicals. Three segregation systems were identified in this study: the UN Dangerous Goods system (which uses physical hazard as the main reason for segregation and has 14 categories), the US CHRIS system (which uses chemical reactivity and has 24 categories) and a third system which uses environmental risks (and has 25 categories). These systems were combined. Merging of each system was initially problematic, but became considerably easier once certain characteristics had been defined (such as flammability or water incompatibility). This gave a final merged incompatibility table containing 100 different segregation groups. This research study showed that it was possible to combine different segregation systems based on different criteria and that more comprehensive segregation systems can be developed. These can be of use in the decision-making process about where groups of chemicals may be used, and during the use of chemicals, where chemicals should not be combined. The use of more comprehensive segregation systems will also assist in developing proper measures for their control. PMID- 11040384 TI - Airblast TNT equivalence for a range of commercial blasting explosives. AB - Results are reported from a programme of work undertaken by the UK Health and Safety Executive to investigate the airblast produced by commercial sector explosives having velocities of detonation (VoD) in the range 2000-8200 m s(-1). The data produced will be useful in evaluating the blast hazards of such explosives in industrial circumstances and also as a means of assessing post accident damage. All of the solid explosive materials studied produced blast waves which ramped up into shock-wave form close to the point of initiation. The dependence of peak overpressure and positive phase impulse on scaled distance is presented and compared to that of TNT. The TNT equivalence (TNT(e)) technique is shown to be applicable to solid phase explosives with a wide range of VoD, although the precise values of TNT(e) vary with distance. PMID- 11040385 TI - Comparison of a reduced explosion model to blast curve and experimental data. AB - This paper discusses a new reduced model for vapor cloud explosions (VCEs) which incorporates some factors found in more complex models such as three-dimensional effects. The common foundation of all VCE analysis models is discussed and a simplified model based on a set of blast curves is reviewed to highlight the differences in the various model assumptions. Output from the reduced model is compared to experimental data and results from the simplified model. It is shown that the reduced model captures critical factors such as cloud shape and flame dynamics. PMID- 11040386 TI - Characterization of metal finishing sludges: influence of the pH. AB - Metal finishing sludges are classified as metal hydroxide hazardous wastes due to the heavy metal release to the environment. This release, commonly determined by compliance lixiviation tests based on the equilibrium conditions at the end of the leaching experiment, is mainly dependent on the pH of the solution. In this work, the leaching behaviour of Cd, Cr, Cu, Fe, Ni, Pb and Zn, of the 32 metal finishing sludges coming from 16 European industrial facilities, and using the distilled water compliance test DIN 38414-S4, have been studied. The concentrations of chromium and copper in the leachates do not follow the solubility evolution of their hydroxide with the pH. The simple assumption of a heavy metal concentration in the leachate directly related to the solubility of the hydroxide is not in good agreement with the experimental results of the distilled water leaching test, probably due to the presence of different species, which can contribute to the metal mobility depending on the sludge composition. An experimental evaluation of the easily available amount of metals in real wastes seems to be necessary for disposal assessment. This paper contains valuable information, from orderly handling metal finishing wastes to the statistical studies of production and management of wastes suggested recently by the Commission of the European Community. PMID- 11040387 TI - Ab initio study of reactions between halogen atoms and various fuel molecules by Gaussian-2 theory. AB - Ab initio calculations by using Gaussian-2 theory have been carried out for the reactions between halogen atoms and various fuel molecules, i.e. fluorine, chlorine, and bromine atoms vs. hydrogen, methane, ethane, ethylene, acetylene, ammonia, silane, dichlorosilane and phosphine. The activation energy for the reaction between a halogen atom and a fuel molecule seems to indicate whether the reaction between the fuel gas and the corresponding halogen gas occurs spontaneously when they are brought into contact to each other at room temperature. PMID- 11040388 TI - The formation of disinfection by-products in water treated with chlorine dioxide. AB - In this study, chlorine dioxide (ClO(2)) was used as an alternative disinfection agent with humic acid as the organic precursor in a natural aquatic environment. The major topics in this investigation consisted of the disinfection efficiency of ClO(2), the formation of disinfection by-products (DBPs), and the operating conditions. The results indicated that the pH value (pH 5-9) did not affect the efficiency of disinfection while the concentration of organic precursors did. The primary DBPs formed were trihalomethanes (THMs) and haloacetic acids (HAAs). The distribution of the individual species was a function of the bromide content. The higher the ClO(2) dosage, the lower the amount of DBPs produced. The amount of DBPs increased with reaction time, with chlorite ions as the primary inorganic by product. PMID- 11040389 TI - Sulphur condensation influence in Claus catalyst performance. AB - The Claus process is an efficient way of removing H(2)S from acid gas streams and this is widely practised in industries such as natural gas processing, oil refining and metal smelting. Increasingly strict pollution control regulations require maximum sulphur recovery from the Claus unit in order to minimise sulphur containing effluent. The most widely used Claus catalyst in sulphur recovery units is non-promoted spherical activated alumina. Properties associated with optimum non-promoted Claus catalyst performance include high surface area, appropriate pore size distribution and enhanced physical properties. The objective of this paper is to outline a procedure in order to estimate Claus catalyst effectiveness after pore plugging due to sulphur condensation. Catalyst deactivation due to pore plugging by sulphur is modelled employing a Bethe lattice and its corresponding performance is described by means of a modified effectiveness factor. Model results show an improvement in the modified effectiveness factor due to modifications in catalyst porous structure. PMID- 11040390 TI - Adsorption studies on Citrus reticulata (fruit peel of orange): removal and recovery of Ni(II) from electroplating wastewater. AB - The ability of fruit peel of orange to remove Zn, Ni, Cu, Pb and Cr from aqueous solution by adsorption was studied. The adsorption was in the order of Ni(II)>Cu(II)>Pb(II)>Zn(II)>Cr(II). The extent of removal of Ni(II) was found to be dependent on sorbent dose, initial concentration, pH and temperature. The adsorption follows first-order kinetics. The process is endothermic showing monolayer adsorption of Ni(II), with a maximum adsorption of 96% at 50 degrees C for an initial concentration of 50 mg l(-1) at pH 6. Thermodynamic parameters were also evaluated. Desorption was possible with 0.05 M HCl and was found to be 95.83% in column and 76% in batch process, respectively. The spent adsorbent was regenerated and recycled thrice. The removal and recovery was also done in wastewater and was found to be 89% and 93.33%, respectively. PMID- 11040391 TI - Electro-migration of nitrate in sandy soil. AB - Migration of nitrate to groundwater has become a serious threat in many agricultural areas. This paper presents the results of experimental laboratory tests studying the nitrate gradient developed in response to an electrical potential. Two systems were tested; the first had no flow (closed system) and the second had flow opposite to the direction of the electrical current. A solution of sodium nitrate in sandy soil was used in both systems. The tests showed that the electro-kinetic process effectively concentrated and retained nitrate close to the anode. The movement of NO(3)(-) through the soil column was significantly influenced by the development of a pH gradient. Statistical analysis was performed to determine best-fit equations relating the nitrate gradient to the electrical input and pH gradient. A simple one-dimensional finite difference model was used to predict the pH gradient developed during the electro-kinetic process. The experimental measurements closely agreed with the predicted spatial and temporal distribution of the nitrate gradient for both closed and open system configurations. PMID- 11040392 TI - An optical technique to determine the total collection efficiency of a cylone. AB - An experimental method is proposed to determine the total collection efficiency of a cyclone. An optical technique is utilized to measure and calculate the total collection efficiency by integrating the area below the size distribution curve upstream and downstream of the cyclone by a particle counter-sizer-velocimeter. The results show that the size distribution upstream and downstream of the cyclone is almost a smooth curve. It could be used to calculate the total collection efficiency. The measured total efficiency by the particle counter sizer-velocimeter is 2-3% higher than that measured by the weighing method. This close agreement between the two methods suggests that the total collection efficiency may be determined by the optical technique. PMID- 11040393 TI - Dissolution rates of limestones of different sources. AB - The dissolution characteristics of limestones from six sources in Taiwan have been studied by using the pH-stat method in a stirred tank at 60 degrees C, pH values of 4 and 6, stirrer speeds of 500-1000 rpm, and a particle size of 75-125 microm aperture width. The dissolution rates of the limestones were found to be controlled by the mass transfer of hydrogen ions with chemical reactions in the liquid film surrounding the limestone particle. The measured value of mass transfer coefficient increases with an increasing pH value and stirrer speed and remains constant with particle size. For the six limestones at the same particle size, the measured dissolution rates per unit area are the same due to the mass transfer control kinetics; however, the time taken to reach a certain fraction of dissolution is proportional to the molar concentration of the soluble species in the limestone and the initial particle size. PMID- 11040394 TI - Removal mechanisms of VOCs in an activated sludge process. AB - This study investigated the factors in plant operating parameters effecting volatile organic compound (VOC) removal and elucidated each individual mechanism in an activated sludge process (ASP). The results suggest that stripping increase with an increase in Henry's law constant with the exception of toluene, which differed because of the effect of biodegradation competition. The emission rate can be controlled by increasing the active biomass concentration (X(b)) in the aeration basin, hydraulic retention time (HRT), and oxygen transfer efficiency (OTE). Increasing X(b) enhances total VOC removal and biodegradation, which reduces the fraction of the stripping removal. Longer HRT can reduce stripping removal significantly if other variables such as oxygen requirement and OTE are held constant. For the same volatile compound, a process with higher air-to-water ratio requires a higher active biomass concentration to reduce stripping removal. Increasing OTE decreases the air-to-water ratio requirement, which reduces the gas-phase transfer of VOCs into the atmosphere. The results show a reasonable agreement between measured and predicted stripping values. PMID- 11040395 TI - Thermodynamic driving forces for PAH isomerization and growth during thermal treatment of polluted soils. AB - For a limiting case of thermodynamic equilibrium, the importance of two classes of thermal chemical reactions that modify the structure and bioactivity of polycyclic aromatic hydrocarbons (PAH) was assessed computationally. These reactions are molecular weight (MW) growth by acetylene addition, and intramolecular rearrangement (isomerization). Temperatures (300-1100 degrees C), and the chemical environment (C(2)H(2)/H(2) molar ratios) were selected for relevancy to thermal treatment of PAH-contaminated soils under oxygen-free conditions. Molecular mechanics methods [MM3(92)] were used to compute thermochemical properties for calculation of equilibrium constants, i.e., heats of formation, standard entropies, and heat capacities for 30 PAH with empirical formulae C(14)H(10), C(16)H(10), C(18)H(10), C(18)H(12), C(20)H(10), and C(20)H(12). Included were 11 PAH containing only six-membered rings and 19 PAH containing both five- and six-membered rings. For each of these PAH the calculations predict that with increasing temperature, isomerization increases the "complexity" of the PAH mixture, i.e., the relative abundance of each PAH isomer in the mixture other than the most stable isomer, increases. Isomerization also partially transforms non-mutagens to mutagens, e.g., pyrene and benzo[e]pyrene to fluoranthene and benzo[a]pyrene, respectively, and partially converts cyclopenta[c, d]pyrene (CPEP) and chrysene, both human cell mutagens, to one and three additional human cell mutagens, respectively. Acetylene addition transforms the non-mutagens phenanthrene and pyrene to the mutagens triphenylene and CPEP, respectively. Some of the predicted PAH have been observed elsewhere among the products of aromatics pyrolysis. This study elucidates PAH reactivity for comparison with measurements, and identifies PAH reactions to be monitored and avoided in soil thermal decontamination and other waste remediation processes. PMID- 11040396 TI - Increase of reactive oxygen (ROS) and nitrogen (RNS) species generated by phagocyting granulocytes related to age. AB - In this present paper the age-induced effect on reactive oxidizing species generated by oxygen (ROS) and nitrogen (RNS) was studied using human phagocyting granulocytes. The ROS and RNS were quantified, respectively, in a chemiluminescence assay and by the measurement of nitrite production. The age induced reactive oxidizing species generation was studied in healthy subjects ranging from 20 to 80 years old, divided into six age groups: group I, 20-29 years old; group II, 30-39 years old; group III, 40-49 years old; group IV, 50-59 years old; group V, 60-69 years old; and group VI, 70-80 years old. Our results demonstrate a parallelism between generation of the ROS and RNS induced by the age. A significant increase of ROS production was observed from 40 years old (age groups III, IV, V and VI while for RNS this increase was observed only from 50 years old (groups IV, V and VI). These data suggest an increase of oxidizing species generation (ROS/RNS) related to age. The increased generation of ROS (40 49 years old) was induced before the increasing of RNS (50-59 years old) and it may have consequences on inflammation and host defences. PMID- 11040397 TI - Postnatal mortality from meningococcal infections during the period 1950-1991 in the US. AB - Research has been conducted on the relationship between postnatal mortality from meningococcal infections and age, using data from the US during the period 1950 1991. The logarithm of mortality caused by meningococcal infections fell linearly with the logarithm of age, during the interval of 1-30 years for men and women in the US. The slope of this straight line is equal to -1. The mortality from meningococcal infections is inversely proportional to the age in the US. The risk of death at age 2 is one half of the risk at age 1, at age 3 it is one third of the risk of death at age 1, etc. up to the age of 30 in the US. The same decline was observed for the risk of death from congenital anomalies and pneumonia. PMID- 11040398 TI - Streptozotocin-induced diabetes and glucocorticoid receptor regulation: tissue- and age-specific variation. AB - Streptozotocin (STZ) -induced diabetic effects were analyzed for glucocorticoid receptor (GR) level and for in vitro activation of GR by specific binding analysis, using [3H]dexamethasone, a synthetic glucocorticoid, and by DNA cellulose and nuclear binding assay, in the liver and kidney of 15- (immature) and 120-day-old (mature) male mice. Comparison of GR level (fmol/mg protein) among the control mice reveals decreased (22-33%) specific binding in the liver and kidney of mature mice compared with immature ones. Scatchard analyses, however, reveal no change in the affinity (K(d)) of receptor at these two ages of mice. STZ-induced diabetes did not alter the level of GR in either of the tissues at both the ages studied. The GR from both the tissues underwent thermal activation, albeit the extent of activation was more pronounced in mature liver compared to immature, with no such difference of activation in the kidney. In diabetic mice, the activation of hepatic GR exhibits reduced DNA cellulose ( approximately 20-23%) and nuclear (24-30%) binding compared to control mice. In contrast, thermal activation of kidney GR does not show marked differences in diabetic mice at either of the ages studied. Cross-mixing experiments (i.e. binding of activated GR from diabetic mice to nuclei of control and vice-versa) performed on the mature liver, indicate receptor specificity. These findings reveal tissue- and age- specific variations in the level of GR that is not influenced under diabetic conditions. However, the activation of hepatic GR is reduced during STZ-induced diabetes that might play a role in controlling glucose homeostasis in diabetic animals. PMID- 11040399 TI - The efficiency of genetic analysis of DNA from aged siblings to detect chromosomal regions implicated in longevity. AB - Studies of the frequencies of different alleles in young adults and aged individuals have implicated several genes, such as ApoE and ACE, in longevity. However such association studies can easily give rise to spurious results through unsuspected population subdivision, and an approach making use of genetic relationships among relatives is desirable. We have studied the effectiveness of non-parametric genetic analysis to detect different types of loci affecting longevity. The non-parametric method has high statistical power to detect infrequent recessive alleles that are required for, or significantly increase the probability of, survival to advanced age. Statistical power is reduced if a proportion of carriers of the alternative allele is allowed to survive. The method is least effective in detecting alleles that occur at low frequency in young individuals and that subsequently experience high mortality, as is the case for carriers of the epsilon4 allele of ApoE. Genotyping errors will also reduce the value of the NPL statistic in a linear fashion with the error rate and the number of loci genotyped. We have also used the method to analyse genotypes of seven highly polymorphic markers near the ApoE gene in a sample of 188 sibships of nonagenarians and centenarians (n=434) and their children (n=124), however no excess sharing of alleles was detected. PMID- 11040400 TI - An hydroalcoholic extract of curcuma longa lowers the apo B/apo A ratio. Implications for atherogenesis prevention. AB - It is generally accepted that free-radical induced blood lipid peroxidation and especially peroxidized LDL play a central role in the pathogenesis of atherosclerosis and related cardiovascular disease. Moreover, recent research highlights the key contribution of apolipoprotein B (apo B) to atherogenesis as the main inductor of one of its earlier steps, i.e. macrophage proliferation. This has led us to investigate the apo B response to a very effective phenolic lipid-antioxidant, namely an hydroalcoholic extract of Curcuma longa, which according to our previous work does not show any toxic effects and decreases the levels of blood lipid peroxides, oxidized lipoproteins and fibrinogen. The present study shows that a daily oral administration of the extract decreases significantly the LDL and apo B and increases the HDL and apo A of healthy subjects. This and recent data on the increased anti-atherogenic action of the physiological antioxidant tocopherol in the presence of phenolic co-antioxidants (which eliminate the tocopheroxyl radical), justifies planned clinical research to test the usefulness of the curcuma extract as a co-antioxidant complement to standard treatments to prevent or retard atherosclerosis. PMID- 11040401 TI - Temporal changes in collagen composition and metabolism during rodent palatogenesis. AB - Cleft lip and palate is a common craniofacial malformation in man. The aetiology is multifactorial and not known. Since collagen is a major structural component of the developing palate, we studied its composition and metabolism during palate shelf formation and elevation in the rat. Palatal shelves were harvested at embryonic days (E) 15, 16 and 17 as well as post-partum. Palatal collagen increased threefold from E15 to E17 and tenfold from E17 to 5-day-old pups. Palatal calcification was seen in the main, post-partum. Collagen cross-linking, which may be important in shelf elevation and union, varied. The concentration of hydroxylysyl-pyridinolone cross-links was greatest prior to shelf elevation, declining thereafter. Similarly, the highest concentration of dihydroxylysinononorleucine was seen at E16 and this supports the concept of a compliant mesenchymal shelf responding to an intrinsic elevating force. We then determined if enzymes responsible for matrix degradation, matrix metalloproteinases (MMP) and the tissue inhibitors of metalloproteinases (TIMPs) altered over the same time periods. MMP-2, and TIMP-1 and TIMP-2 were identified by gelatin zymography and reverse zymography, respectively. MMP-3 activity was determined with a fluorogenic substrate assay. TIMP-1, TIMP-2 and MMP-3 levels remained constant from E15 to E17. The MMP-2 levels showed a significant elevation from E15 to E16 and E16 to E17. This suggests the regulation of extracellular matrix is likely to be of importance in palate morphogenesis. PMID- 11040402 TI - Beta-amyloid-induced activation of caspase-3 in primary cultures of rat neurons. AB - It is known that beta-amyloid peptide (Abeta) contributes to the neurodegeneration in Alzheimer's disease (AD) and operates through activation of an apoptotic pathway. Apoptotic signal is driven by a family of cysteine proteases called caspases. The beta-amyloid precursor protein (APP) is directly and efficiently cleaved by caspases during apoptosis, resulting in elevated beta amyloid peptide formation. Cerebellar neurons from rat pups were treated with the aged Abeta(25-35) at 1 and 5 microM and fluorescence assays of caspase activity performed over 4 days. We observed an increase in caspase activity after 48 h treatment in both 1 and 5 microM treated cells, then (72-96 h) caspase activity decreased to control values. The data presented support the hypothesis that Abeta(25-35)-induced apoptosis is mediated by the activation of Caspase-3 and that this is a transient effect. PMID- 11040403 TI - Regulatory role of extracellular matrix proteins in neutrophil respiratory burst during aging. AB - Neutrophil respiratory burst was assessed on plates coated with fibronectin (FN) or laminin (LM), both used at dosages inhibiting polystyrene-triggered cell activation in young healthy volunteers. Under these conditions, a low, yet significant, spontaneous superoxide anion (O(2)(-)) production, matching with enhanced levels of basal adherence, was detected in FN-plated neutrophils of elderly donors. In contrast, although neutrophil stimulation with tumor necrosis factor (TNF)-alpha, granulocyte macrophage-colony stimulating factor (GM-CSF), fMLP or phorbol myristate acetate (PMA) gave rise to a massive and prolonged FN primed O(2)(-) release, a significant impairment of oxidative response occurred in the aged group as a result of GM-CSF or fMLP cell challenge. Such an effect was not associated to an age-related imbalance of stimulant-triggered neutrophil adhesiveness to FN, even though a larger contribution of CD18-dependent versus CD18-independent pathways was observed in old as compared to young individuals. Notably, within the aged group, anti-CD18 monoclonal antibody cell pretreatment resulted in a higher suppression of FN-primed O(2)(-) release following TNF-alpha with respect to GM-CSF stimulation, thus implying that an agonist-related defect of the coupling between beta2 integrin-dependent adhesive and oxidative events is likely to occur as a feature of age. All physiological mediators failed to activate the respiratory burst of neutrophils plated on LM-coated wells in both young and aged donors. This effect appears to be the result of an active process, since neutrophils from either group of subjects adhered to LM-coated surfaces and LM inhibited in a dose-dependent manner the FN-priming effect on neutrophil O(2)( ) production. All together the findings provide additional evidence for an imbalance of neutrophil-mediated functions in the elderly. PMID- 11040404 TI - Erratum to 'Electrophysiological analysis of NMDA receptor subunit changes in the aging mouse cortex'. PMID- 11040405 TI - Experimental microneurosurgery of the trigeminal ganglion and ophthalmic maxillary nerve in the rat: subtemporal fossa approach. AB - In the researches of the innervation relationship between trigeminal ganglion and a particular structure in the head, it is usually necessary to apply neural tracers into the ganglion for anterograde nerve tracing study or perform bilateral trigeminal nerve transecting for degeneration study. A common surgical approach for exposing these structures in the rat was to remove a piece of skull and a portion of brain. While investigating the innervation of rat's pineal gland from its trigeminal ganglion, we used a subtemporal fossa approach, whereby the mortality of the animal was remarkably reduced and the contamination of the intracranial structures by the tracer was proved to be least. This is the first description of bilateral surgeries of the trigeminal ganglion and trigeminal nerve in rat using extracranial approach. Detailed surgical procedures were presented and their advantages discussed. PMID- 11040406 TI - A system for simultaneous multiple subject, multiple stimulus modality, and multiple channel collection and analysis of sensory evoked potentials. AB - A system has been developed for collecting sensory evoked potentials simultaneously from multiple channels for multiple subjects at up to 80 kHz sample rate per channel. Sample rates up to 200 kHz are available for four or less chambers and a single channel per chamber. A variety of visual, somatosensory, and auditory stimuli may be presented singly or simultaneously. Collected waveforms are associated with searchable text (metadata) to allow convenient selection from a relational database. Multiple waveforms can then be easily grouped for analysis and processed. Results can be exported to other software for further graphics or statistical processing. Scripting and event logging are available to provide automation and improve data confidence. Sample data are presented from control animals for each of the sensory modalities for comparison with historical data collected from other systems. PMID- 11040407 TI - Video analysis of standing--an alternative footprint analysis to assess functional loss following injury to the rat sciatic nerve. AB - The rat sciatic nerve is a well-established animal model for the study of recovery from peripheral nerve injuries. Footprint analysis is the most widely used non-invasive method of measuring functional recovery after injury in this model. We describe a new alternative video analysis of standing (or static footprint video analysis) to assess functional loss following injury to the rat sciatic nerve, during animal standing or periodic rest on a flat transparent surface. We found good correlation between video recording during standing and dynamic ink track footprint parameter measurements for both 1-5 and injured 2-4 toe spreads only. Reproducibility for these three parameters was also better using the video method. Uninjured 2-4 toe spread by video showed a poor correlation and similar reproducibility as compared with ink. However, both print length parameters measured by video had poorer correlation and greater variability, particularly the print length factor (PLF) was weakly correlated with that determined by ink. Contribution of the footprint factors on the estimated functional loss has also changed in conditions during standing. It was most prominent for the 1-5 toe spread factor (TSF), near marginal for the 2-4 or intermediary toe spread factor (ITF), and weak, statistically insignificant for the PLF. Thus, the introduction of a new functional loss index, or so-called static sciatic index (SSI), and its estimating formula was mandatory. Moreover, using a simple ratio of injured/uninjured 1-5 video toe spread as a substitute for the SSI, we could achieve considerable simplification of the method without any significant loss of accuracy. Our video analysis of standing is technically easier to perform than the corresponding footprint video analysis during walking, but still preserves all advantages of video versus conventional ink track method, i.e. there are few non-measurable footprints, better repeatability, high accuracy and more precise quantification of the degree of functional loss after sciatic nerve injury in the rat. PMID- 11040408 TI - The plus-maze discriminative avoidance task: a new model to study memory-anxiety interactions. Effects of chlordiazepoxide and caffeine. AB - The plus-maze discriminative avoidance paradigm is a new animal model of learning/memory that provides simultaneous information about anxiety. Mice are conditioned to choose between the two enclosed arms (in one of which light and noise are presented as aversive stimuli) while avoiding the two open arms of the apparatus. The test has the advantage of measuring, at the same time and in the same animals, learning/memory (by the percent time spent in aversive enclosed arm - PTAV) and anxiety (by the percent time spent in the open arms - PTO). The effects of chlordiazepoxide and caffeine on learning/memory and anxiety of mice tested in this paradigm were investigated. Chlordiazepoxide (5 mg/kg) significantly increased and caffeine (20 mg/kg) significantly decreased PTO during the training session, suggesting an anxiolytic and an anxiogenic effect, respectively. In the test session, chlordiazepoxide- or caffeine-treated mice presented higher PTAV, suggesting amnestic effects. Given together, chlordiazepoxide plus caffeine did not alter PTO, and the amnesic effect produced by each drug was no longer observed. It is concluded that learning/memory depends on an optimum emotional level. The plus-maze discriminative avoidance model appears to be a useful test to investigate this critical relationship between learning/memory and anxiety. PMID- 11040409 TI - Comparison by microdialysis of striatal L-DOPA after its systemic administration in rats with probes implanted acutely or through a guide cannula. AB - Different methods of microdialysis probe implantation are utilized according to the purpose and needs of each particular study. However, very few experiments have systematically examined whether these different techniques have an impact on the obtained data. In the present study we examined the influence of two different microdialysis methods - acute probe implantation vs. insertion into a preimplanted guide cannula - on the striatal extracellular availability of systemically administered L-DOPA. Furthermore, we monitored the effects of L-DOPA administration on dopamine and its metabolite 3,4-dihydroxyphenylacetic acid (DOPAC). In rats that received a guide cannula 4 days prior to probe insertion and microdialysis, extracellular L-DOPA concentrations increased to concentrations that were about nine times higher than in rats with acute implantation of a microdialysis probe. Extracellular DOPAC concentrations were also higher in the chronic preparations but dopamine concentrations showed no differences between groups. Our results suggest that the observed differences may be due to inflammatory disruption of the BBB following chronic implantation of a guide cannula. PMID- 11040410 TI - Multi-lipofection efficiently transfected genes into astrocytes in primary culture. AB - This study demonstrated that liposome-mediated transfection - lipofection - is suitable for delivering genes into astrocytes. By repeatedly lipofecting the same astrocyte cultures, a process we call multi-lipofection, the transfection efficiency of the beta-galactosidase (beta-gal) gene was improved from 2.6+/-0.6 to 17. 4+/-1.1%. This is the highest efficiency ever reported in gene-transfer with Lipofectin(R) in a primary culture of mouse cerebral cortical astrocytes. Furthermore, multi-lipofection did not cause observable disturbance to astrocytes as indicated by insignificant changes in the glial fibrillary acidic protein content in the cultures. In order to demonstrate that the transfected gene achieved a physiologically relevant expression level, a plasmid containing the pEF-hsp70 protein gene was lipofected into astrocytes. This produced colonies of astrocytes showing an increased resistance to heat-induced cell death. A similar experiment was performed with the glial-derived neurotrophic factor (GDNF) gene. Control astrocytes had no detectable GDNF. In the transfected astrocytes, the GDNF protein could be identified intracellularly by immunocytochemistry. Western blot analysis revealed, as compared to astrocytes with one lipofection, a 2.9 fold increase of GDNF with four lipofections. GDNF remained detectable in astrocytes 2 weeks after four lipofections. Thus, multi-lipofection provides a mild and efficient means of delivering foreign genes into astrocytes in a primary culture, making astrocytes good candidate vehicle cells for gene/cell therapy in the CNS. PMID- 11040411 TI - Automated difference image analysis of lamellar ruffling: effect of temperature change on human SH-SY5Y neuroblastoma cells. AB - Quantitation of cell movement and lamellar extension is critical to the study of growth factors, chemotactic agents and signaling cascades. Many studies are conducted by examining the size or number of lamellae in static images of cells. However, these methods do not quantify lamellar behavior over time and may overlook important changes in lamellar function. Most presently available methods for analyzing dynamic aspects of lamellar function examine changes in a cell's 2 dimensional perimeter and are best suited to the analysis of flattened lamellae. However, some cells generate 3-dimensional lamellar ruffles whose behavior is not readily detected using these methods. In the present study we analyze temperature dependent ruffling of human SH-SY5Y neuroblastoma cells using automated digital subtraction of time-lapse images and quantitation of the resultant 'difference' image, and compare results obtained using this and other approaches. We report that ruffling behavior of SH-SY5Y cells is measurably altered by temperature changes of as little as 1 degrees C, and that these changes are best detected using difference image analysis. Our studies indicate that temperature is a critical variable in studies of SH-SY5Y behavior and that difference image analyses may be an important complement to other methods in the study of lamellar ruffling. PMID- 11040412 TI - Passive and active place avoidance as a tool of spatial memory research in rats. AB - A modified model of the arena described by Bures et al. (Bures J, Fenton AA, Kaminsky Y, Zinyuk L. Place cells and place navigation, Proc. Natl. Acad. Sci. USA. 1997a;94:343-350) was applied to the place learning of adult male rats in two different avoidance paradigms. In the passive avoidance task rats exploring a stationary circular arena had to avoid a 60 degrees sector entering of which was punished by mild footshocks. Intramaze as well as extramaze cues could be used for adequate solution of this task. In the active avoidance paradigm rats were trained to avoid a room frame defined sector (e.g. North-East) of a slowly rotating arena the movement of which forced the animals to rely on extramaze cues and to ignore intramaze information. Rats had to find an active solution of the task since otherwise they were passively transported into the room frame defined punished zone. The suitability of these tasks for testing spatial abilities of rats is discussed. PMID- 11040413 TI - Calcium imaging in live rat optic nerve myelinated axons in vitro using confocal laser microscopy. AB - Intracellular Ca(2+) plays a major role in the physiological responses of excitable cells, and excessive accumulation of internal Ca(2+) is a key determinant of cell injury and death. Many studies have been carried out on the internal Ca(2+) dynamics in neurons. In constrast, there is virtually no such information for mammalian central myelinated axons, due in large part to technical difficulty with dye loading and imaging such fine myelinated structures. We developed a technique to allow imaging of ionized Ca(2+) in live rat optic nerve axons with simultaneous electrophysiological recording in vitro at 37 degrees C using confocal microscopy. The K(+) salt of the Ca(2+)-sensitive indicator Oregon Green 488 BAPTA-2 and the Ca(2+)-insensitive reference dye Sulforhodamine 101 were loaded together into rat optic nerves using a low Ca(2+)/low-Na(+) solution. Axonal profiles, confirmed immunohistochemically by double staining with neurofilament-160 antibodies, were clearly visualized by S101 fluorescence up to 800 microm from the cut ends. The Ca(2+) signal was very low at rest, just above the background fluorescence intensity, indicating healthy tissue, and increased significantly after caffeine (20 mM) exposure designed to release internal Ca(2+) stores. The health of imaged regions was further confirmed by a virtual absence of spectrin breakdown, which is induced by calpain activation in damaged CNS tissue. Red and green fluorescence decayed to no less than 70% of control after 60 min of recording at 37 degrees C, with the green:red fluorescence ratio increasing slightly by 21% after 60 min. Electrophysiological responses recorded simultaneously with confocal images remained largely stable as well. PMID- 11040414 TI - A robotic manipulator for the characterization of two-dimensional dynamic stiffness using stochastic displacement perturbations. AB - Experimental techniques for estimating the two-dimensional dynamic stiffness of the human arm over a wide range of conditions have been developed. A robotic manipulator has been developed to create loads against which subjects perform various tasks and also to impose perturbations onto the endpoint of the arm to allow estimation of its mechanical properties. The manipulator can produce static endpoint forces exceeding 220 N in any direction in its plane of motion, and this plane can be vertically translated and tilted over wide ranges to study arm dynamic stiffness in many functionally relevant planes. It can impose stochastic position and force perturbations whose bandwidth exceeds that of the arm. These random perturbations avoid undesirable volitional reactions and allow the efficient estimation of stiffness dynamics using experimental trials of short duration. The ability of this manipulator to characterize inertial-viscoelastic systems was tested using several two-dimensional physical systems whose properties were independently characterized. The endpoint dynamic stiffness properties of a human arm were estimated as an example of the use of the manipulator in studying upper limb mechanical properties. The system properties characterized by these methods will be useful in probing normal neural arm control strategies and in developing rehabilitation interventions to improve arm movements in disabled individuals. PMID- 11040415 TI - Primary neural precursor cell expansion, differentiation and cytosolic Ca(2+) response in three-dimensional collagen gel. AB - To investigate the ability to culture neural precursor cells in a three dimensional (3D) collagen gel, neuroepithelial cells were isolated from embryonic day 13 rat cortex, dispersed within type I collagen and maintained for up to 30 days in vitro. Cultured in Neuorobasal medium supplemented with B27 containing basic fibroblast growth factor, the collagen-entrapped precursor cells actively expanded and formed clone-like clusters. Many cells in the center of the cluster were proliferating as revealed by 5-bromo-2'-deoxyuridine uptake. Some cells began to migrate away from the center at 5 days and were labeled by either neuronal marker neuron-specific beta-tubulin (TuJ1) or astrocytic marker glial fibrillary acidic protein. The differentiated neurons (TuJ1(+)) exhibited characteristic cytosolic Ca(2+) oscillations in response to excitatory neurotransmitter glutamate. These findings suggest the suitability of the 3D culture system for the proliferation and differentiation of neural precursor cells. PMID- 11040416 TI - Spatial hemineglect in humans. AB - The paper reviews the main findings of studies of hemispatial neglect after acquired brain lesions in people. The behavioral consequences of experimentally induced lesions in animals and electrophysiological studies, which shed light on the nature of the disorder, are briefly considered. Neglect is behaviorally defined as a deficit in processing or responding to sensory stimuli in the contralateral hemispace, a part of the own body, the part of an imagined scene, or may include the failure to act with the contralesional limbs despite intact motor functions. Neglect in humans is frequently encountered after right parieto temporal lesions and leads to a multicomponent syndrome of sensory, motor and representational deficits. Relevant findings relating to neglect, extinction and unawareness are reviewed and include the following topics: etiological and anatomical basis, recovery; allocentric, egocentric, object-centered and representational neglect; motor neglect and directional hypokinesia; elementary sensorimotor and associated disorders; subdivisions of space and frames of reference; extinction versus neglect; covert processing of information; unawareness of deficits; human and animal models; effects of sensory stimulation and rehabilitation techniques. PMID- 11040417 TI - Neuroprotection by estradiol. AB - This review highlights recent evidence from clinical and basic science studies supporting a role for estrogen in neuroprotection. Accumulated clinical evidence suggests that estrogen exposure decreases the risk and delays the onset and progression of Alzheimer's disease and schizophrenia, and may also enhance recovery from traumatic neurological injury such as stroke. Recent basic science studies show that not only does exogenous estradiol decrease the response to various forms of insult, but the brain itself upregulates both estrogen synthesis and estrogen receptor expression at sites of injury. Thus, our view of the role of estrogen in neural function must be broadened to include not only its function in neuroendocrine regulation and reproductive behaviors, but also to include a direct protective role in response to degenerative disease or injury. Estrogen may play this protective role through several routes. Key among these are estrogen dependent alterations in cell survival, axonal sprouting, regenerative responses, enhanced synaptic transmission and enhanced neurogenesis. Some of the mechanisms underlying these effects are independent of the classically defined nuclear estrogen receptors and involve unidentified membrane receptors, direct modulation of neurotransmitter receptor function, or the known anti-oxidant activities of estrogen. Other neuroprotective effects of estrogen do depend on the classical nuclear estrogen receptor, through which estrogen alters expression of estrogen responsive genes that play a role in apoptosis, axonal regeneration, or general trophic support. Yet another possibility is that estrogen receptors in the membrane or cytoplasm alter phosphorylation cascades through direct interactions with protein kinases or that estrogen receptor signaling may converge with signaling by other trophic molecules to confer resistance to injury. Although there is clear evidence that estradiol exposure can be deleterious to some neuronal populations, the potential clinical benefits of estrogen treatment for enhancing cognitive function may outweigh the associated central and peripheral risks. Exciting and important avenues for future investigation into the protective effects of estrogen include the optimal ligand and doses that can be used clinically to confer benefit without undue risk, modulation of neurotrophin and neurotrophin receptor expression, interaction of estrogen with regulated cofactors and coactivators that couple estrogen receptors to basal transcriptional machinery, interactions of estrogen with other survival and regeneration promoting factors, potential estrogenic effects on neuronal replenishment, and modulation of phenotypic choices by neural stem cells. PMID- 11040418 TI - Rapid plasticity of dendritic spine: hints to possible functions? AB - Contrary to a century-old belief that dendritic spines are stable storage sites of long term memory, the emerging picture from a recent flurry of exciting observations using novel high resolution imaging methods of living cells in culture is that of a dynamic structure, which undergoes fast morphological changes over periods of hours and even minutes. Concurrently, the nature of stimuli which cause formation or collapse of dendritic spines has changed from a mysterious Hebbian-governed plasticity producing stimulus to the more trivial activation of the synapse by strong/weak stimulation. The molecular mechanisms underlying spine plasticity are beginning to emerge; the role of presynaptic and/or postsynaptic activity, genetic, central or local factors in the formation and retraction of spines are currently being analyzed. A common mechanism for both, formation/elongation and pruning/retraction of spines, involving changes in intracellular calcium concentration ([Ca(2+)](i)), is emerging. It appears that [Ca(2+)](i) is related to changes in spines in a bell shape form: lack of synaptic activity causes transient outgrowth of filopodia but eventual elimination of spines, a moderate rise in [Ca(2+)](i) causes elongation of existing spines and formation of new ones, while a massive increase in [Ca(2+)](i) such as that seen in seizure activity, causes fast shrinkage and eventual collapse of spines. Nuclear signals (e.g. CREB), activated by an increase in [Ca(2+)](i), are involved in the central regulation of spine formation, while spine shrinkage and elongation are probably triggered by local [Ca(2+)](i) changes. This hypothesis provides a parsimonious explanation for conflicting reports on activity-dependent changes in dendritic spine morphology. Still, the many differences between cultured neurons, with which most of current studies are conducted, and the neuron in the real brain, require a cautious extrapolation of current assumptions on the regulation of spine formation. PMID- 11040419 TI - Brain-derived neurotrophic factor in the control human brain, and in Alzheimer's disease and Parkinson's disease. AB - Brain-derived neurotrophic factor (BDNF) is a small dimeric protein, structurally related to nerve growth factor, which is abundantly and widely expressed in the adult mammalian brain. BDNF has been found to promote survival of all major neuronal types affected in Alzheimer's disease and Parkinson's disease, like hippocampal and neocortical neurons, cholinergic septal and basal forebrain neurons, and nigral dopaminergic neurons. In this article, we summarize recent work on the molecular and cellular biology of BDNF, including current ideas about its intracellular trafficking, regulated synthesis and release, and actions at the synaptic level, which have considerably expanded our conception of BDNF actions in the central nervous system. But our primary aim is to review the literature regarding BDNF distribution in the human brain, and the modifications of BDNF expression which occur in the brain of individuals with Alzheimer's disease and Parkinson's disease. Our knowledge concerning BDNF actions on the neuronal populations affected in these pathological states is also reviewed, with an aim at understanding its pathogenic and pathophysiological relevance. PMID- 11040420 TI - Evidence for the involvement of bacterial superantigens in psoriasis, atopic dermatitis, and Kawasaki syndrome. AB - A growing body of evidence implicates streptococcal and staphylococcal superantigens in the development of psoriasis, atopic dermatitis and Kawasaki syndrome. In each of these illnesses, an abnormal state of immunologic activity is observed. Superantigens, which have a unique ability to activate large numbers of lymphocytes, are likely to contribute to these disorders in a number of ways. The demonstrated activities of bacterial superantigens include increasing the number of circulating lymphocytes, with activation of autoreactive subsets, upregulation of tissue homing receptors on circulating lymphocytes, and local activation of immune cells within affected tissues. Through these and other mechanisms, superantigens have a proven ability to induce high levels of inflammatory cytokines and/or initiate autoimmune responses that contribute to the development of skin and vascular disorders. Though development of the illnesses discussed in this review are highly complex processes, superantigens may well play a critical role in their onset or maintenance. Understanding superantigen function may elucidate potential therapeutic strategies for these disorders. PMID- 11040421 TI - Comparison of the ADP-ribosylating thermozyme from Sulfolobus solfataricus and the mesophilic poly(ADP-ribose) polymerases. AB - The poly(ADP-ribose) polymerase-like thermozyme purified from Sulfolobus solfataricus was characterised with respect to some physico-chemical properties. The archaeal protein exhibited a scarce electrophoretic mobility at both pH 2.9 and pH 7.5. Determination of the isoelectric point (pI=7.0-7.2) allowed us to understand the reason for the limited migration at pH 7.5, while amino acid composition analysis showed a moderate content of basic residues, which reduced mobility at pH 2.9. With respect to the charge, the archaeal enzyme behaved differently from the eukaryotic thermolabile poly(ADP-ribose) polymerase, described as a basic protein (pI=9.5). Well known inhibitors of the mesophilic polymerase like Zn(2+), nicotinamide and 3-aminobenzamide exerted a smaller effect on the enzyme from S. solfataricus, reducing the activity by at most 50%. Mg(2+) was a positive effector, although in a dose-dependent manner. It influenced the fluorescence spectrum of the archaeal protein, whereas NaCl had no effect. PMID- 11040422 TI - DNA sequence of the insertional hot spot of Tn916 in the Clostridium difficile genome and discovery of a Tn916-like element in an environmental isolate integrated in the same hot spot. AB - Tn916 is a broad host range tetracycline resistance conjugative transposon. In most bacteria, this element enters the bacterial genome at multiple sites. However, in Clostridium difficile, the element has a strong hot spot when introduced by filter mating from Bacillus subtilis. In this work, the DNA sequence of the preferred insertion site (att916) was obtained. An environmental isolate of C. difficile was also discovered which contained an element indistinguishable from Tn916, Tn916CD. Tn916CD was found integrated at att916. PMID- 11040423 TI - Biotransformation of the isoflavonoids biochanin A, formononetin, and glycitein by Eubacterium limosum. AB - Eubacterium limosum (ATCC 8486), a strict anaerobe from the human intestinal tract that is capable of O-demethylation of several compounds, was tested for the ability to metabolize three methoxylated isoflavonoids, biochanin A, formononetin, and glycitein. High-performance liquid chromatography elution profiles of metabolites produced from biochanin A, formononetin, and glycitein showed peaks that had identical retention times to authentic genistein, daidzein, and 6,7,4'-trihydroxyisoflavone, respectively. The metabolites were identified, using an on line liquid chromatography-electrospray mass spectrometer. E. limosum produced 61.4 microM of genistein and 13.2 microM of daidzein from 100 microM of biochanin A and formononetin, after 26 days incubation. O-demethylase activity is cell-associated and was not detected in the extracellular fraction of bacterial culture. This is the first study in which conversion of biochanin A, and formononetin to more potent phytoestrogens by a bacterium has been shown. PMID- 11040424 TI - Complement activation in SCID and nude mice is related to severity of tissue inflammation in the Candida mastitis model. AB - A small animal model of localised candidiasis is needed for the evaluation of new antifungal compounds. Mammary glands of immunocompetent (BALB/cJ) and immunodeficient (SCID and athymic nude) mice were infected with a wild-type of Candida albicans. Some of the animals were treated with amphotericin B (AmB) while others were treated with saline and acted as controls. The histologic changes of infected mammary gland tissues and a number of other organs were evaluated. Complement (C) activation was analysed by immunoelectrophoretic quantification of molecules with C3c epitopes (C3, C3b, iC3b, and C3c) in serum. In all animals the organisms were confined to the mammary glands. Serum C3c levels were significantly higher (P<0.05) in infected untreated BALB/cJ and SCID mice, which also had severe mammary gland tissue inflammation, compared with control mice treated with AmB (4 mg kg(-1) i.p. once daily for 4 days). Both treated and control nude mice showed less tissue inflammation compared to BALB/cJ and SCID mice, and revealed insignificant activation of the complement system. It is concluded that innate immune response is important in the control of candidiasis and that the murine mastitis model is useful for immunopathological studies as well as evaluation of potential antifungal compounds. PMID- 11040425 TI - The yabG gene of Bacillus subtilis encodes a sporulation specific protease which is involved in the processing of several spore coat proteins. AB - The synthesis and proteolysis of the spore coat proteins, SpoIVA and YrbA, of Bacillus subtilis were analyzed using antisera. Almost no intact full-length proteins of either type were extracted from wild-type spores, while yabG mutant spores contained intact SpoIVA and YrbA proteins. We purified recombinant YrbA and YabG proteins from Escherichia coli transformants and found that YrbA was cleaved to the smaller moiety in the presence of YabG in vitro. These observations indicate that YabG is a protease involved in the proteolysis and maturation of SpoIVA and YrbA proteins, conserved with the cortex and/or coat assembly by B. subtilis. PMID- 11040426 TI - Aspergillus fumigatus alb1 encodes naphthopyrone synthase when expressed in Aspergillus oryzae. AB - Conidial pigment biosynthesis is an important virulence factor in Aspergillus fumigatus, a human fungal pathogen. Involvement of DHN-melanin pathway in the biosynthesis of A. fumigatus conidial pigment implies that the Alb1p polyketide synthase (PKS) is a 1,3,6, 8-tetrahydroxynaphthalene (T4HN) synthase. The Alb1p, however, shows higher sequence similarity to a naphthopyrone synthase than to a T4HN synthase. To clarify the function of Alb1p, the alb1 gene was overexpressed in a heterologous host Aspergillus oryzae. The Alb1p PKS product in this heterologous expression system was identified as heptaketide naphthopyrone instead of pentaketide T4HN. The data suggest that Alb1p is a naphthopyrone synthase. PMID- 11040427 TI - Random amplification of polymorphic DNA versus pulsed field gel electrophoresis of SmaI DNA macrorestriction fragments for typing strains of vancomycin-resistant enterococci. AB - Genetic typing of vancomycin-resistant enterococci (VRE) can be performed using a variety of methods, but comparative analyses of the quality of these methods are still relatively scarce. We here compare random amplification of polymorphic DNA (RAPD) analysis with pulsed field gel electrophoresis (PFGE) of DNA macrorestriction fragments as examples of two of the recent and well-accepted molecular typing methods. For the latter method, empirical guidelines for the interpretation of the DNA fingerprints have been proposed in the international literature. Based on our experimental analyses, we define similar criteria for RAPD fingerprinting. A collection of 100 strains of VRE, comprising Enterococcus faecium, Enterococcus faecalis, Enterococcus avium, Enterococcus gallinarum and Enterococcus casseliflavus, was assembled. Fifty isolates were Dutch, another 50 were isolated in the UK. Strains were selected on the basis of previously determined putative identity, close relatedness or uniqueness. The strains were analysed using well-standardised RAPD and PFGE protocols. Resulting fingerprints were interpreted with computerised methods involving band positioning and we show that typing of VRE by PFGE and RAPD generates highly congruent DNA fingerprint clustering. When the proposed international criteria for interpretation of PFGE fingerprints were applied in our case, 86% PFGE homology as discriminating value between close relatedness and uniqueness, a 75% homology cut-off for the comparison of the RAPD-generated DNA fingerprints revealed essentially identical strain clusters. As a spin-off it is revealed that strains from the different species can be efficiently discriminated, that strains from the UK and The Netherlands form separate clusters and that strains from veterinary origin can be identified separately as well. PMID- 11040428 TI - Location of repeat elements in glucansucrases of Leuconostoc and Streptococcus species. AB - Glucosyltransferases of oral streptococci, dextransucrases and alternansucrase of Leuconostoc mesenteroides, collectively referred to as glucansucrases, are large extracellular enzymes that synthesise glucans with a variety of structures and properties. A characteristic of all these glucansucrases is the possession of a C terminal domain consisting of a series of tandem amino acid repeats. These repeat units are thought to interact with glucan but closely resemble the cell wall binding domain motif found in choline binding proteins in Streptococcus pneumoniae and surface-located proteins in a range of other bacteria. Analysis of dextransucrase and alternansucrase sequences has now shown that they also contain these repeat motifs in the N-terminal region, raising questions about their evolutionary origin and functional importance. PMID- 11040429 TI - Distribution of ecto 5'-nucleotidase on Mycoplasma species associated with arthritis. AB - The enzyme ecto 5'-nucleotidase (5'N) was found to be active on 8/14 strains of Mycoplasma fermentans, K(m) (+/-S.D.) 3.8+/-2.8 microM 5'-AMP, and on the type strain of Mycoplasma pulmonis, K(m) 0.63 microM 5'-AMP. The six M. fermentans strains lacking 5'N activity were related by restriction fragment length polymorphism typing. At pH 8.5, the type strains of Mycoplasma arthritidis, Mycoplasma buccale and Ureaplasma urealyticum showed a relatively non-specific phosphatase activity against 5'-AMP but no activity was shown by the type strains of Mycoplasma genitalium, Mycoplasma hominis, Mycoplasma orale, Mycoplasma penetrans, Mycoplasma pneumoniae and Mycoplasma salivarium at this pH. M. fermentans has been reported from rheumatoid joints, which show a raised 5'N activity on their synovial cells and in their fluid which may be associated directly or indirectly with the mycoplasma. PMID- 11040430 TI - Sequencing and characterization of a novel serine metalloprotease from Burkholderia pseudomallei. AB - Burkholderia pseudomallei, a Gram-negative bacterium is found in the soil and water, mainly in Southeast Asia and Northern Australia. It is responsible for melioidosis in human and animals. The bacteria produce several potential virulent factors such as extracellular protease, hemolysin, lipase and lecithinase. The isolation of virulence genes and the study of their functions will contribute to our understanding of bacterial pathogenesis. Previous studies have implicated protease as a contributing virulence factor in the pathogenesis of some bacteria. Three out of 5000 clones screened from a genomic DNA library of B. pseudomallei were found to express protease activity. The clones were found to have the same sequence. The nucleotide sequence revealed an open reading frame (designated as metalloprotease A, mprA) encoding a 500-amino acid protein, MprA, with an estimated molecular mass of 50241 Da. The predicted amino acid sequence shares homology with the subtilisin family of serine proteases. PMID- 11040431 TI - Structural variation in the 16S-23S rRNA intergenic spacers of Vibrio parahaemolyticus. AB - The 16S-23S rRNA intergenic spacers (IGS) of Vibrio parahaemolyticus were PCR amplified and cloned with pT7Blue T vector. A total of six clones were isolated dependent on size difference. The clones were different with respect to both the number and the composition of the tRNA genes included, and were designated IGS-0, IGS-E, IGS-IA, IGS-AE, IGS-EKV and IGS-EKAV. IGS-EKAV included the cluster of tRNA(Glu)-tRNA(Lys)-tRNA(Ala)-tRNA(Val); IGS-EKV, tRNA(Glu)-tRNA(Lys)-tRNA(Val); IGS-AE, tRNA(Ala)-tRNA(Glu); IGS-IA, tRNA(Ile)-tRNA(Ala); and IGS-E, tRNA(Glu). IGS-0 had no tRNA gene. Some similarities were found in the nucleotide sequence of the non-coding regions flanked by the tRNA genes. The structure difference found in the spacers is meaningful for elucidating the evolutionary line of each ribosomal RNA operon and the profile is applicable as a molecular marker of the bacterium. PMID- 11040432 TI - Effect of enterocin CRL35 on Listeria monocytogenes cell membrane. AB - The antimicrobial peptide Enterocin CRL35, a class II bacteriocin, produces at high concentrations (8 microg ml(-1)) localized holes in the wall and cellular membrane of Listeria monocytogenes, reflected in the efflux of macromolecules such as proteins and other ultraviolet-absorbing materials. At lower concentrations (0.5 microg ml(-1)), neither ultra structural changes nor macromolecules efflux were observed, however potassium and phosphate ions were released, dissipating the proton motive force. As a result the bacteria were killed. PMID- 11040433 TI - Zinc uptake, oxidative stress and the FNR-like proteins of Lactococcus lactis. AB - Lactococcus lactis ssp. cremoris MG1363 contains two FNR homologues, FlpA and FlpB, encoded by the distal genes of two paralogous operons (orfX(A/B)-orfY(A/B) flpA/B). An flpA flpB double mutant strain is hypersensitive to hydrogen peroxide and has a depleted intracellular Zn(II) pool. The phenotypes of the flp mutant strains suggest that FlpA and FlpB control the expression of high and low affinity ATP-dependent Zn(II) uptake systems, respectively. Plate tests revealed that expression from a orfX(B)::lac reporter was activated by Cd(II), consistent with other Zn(II)-regulated systems. The link between a failure to acquire Zn(II) and hypersensitivity to oxidative stress suggests that Zn(II) may be required to protect vulnerable protein thiols from oxidation. PMID- 11040434 TI - Glutamine synthetase gene expression at elevated hydrostatic pressure in a deep sea piezophilic Shewanella violacea. AB - A glutamine synthetase gene (glnA) was isolated from a deep-sea piezophilic bacterium, Shewanella violacea strain DSS12. A 7.5-kb SacI fragment containing the complete glnA gene was cloned and sequenced. The glnA gene was found to encode a protein consisting of 469 amino acid residues, showing 75.0% identity to the glutamine synthetase of Escherichia coli. Primer extension analyses revealed two transcription initiation sites in glnA and expression from each site was positively regulated by pressure. Putative promoters recognized by sigma(70) and sigma(54) were identified in the region upstream of glnA. An electrophoretic mobility shift assay demonstrated that S. violacea sigma(54) specifically binds to the promoter region of glnA, suggesting that sigma(54) may play an important role in pressure-regulated transcription in this piezophilic bacterium. PMID- 11040435 TI - Effect of the adhesive antibiotic TA on adhesion and initial growth of E. coli on silicone rubber. AB - Catheter-associated urinary tract infection is the most common nosocomial infection, and contributes to patient morbidity and mortality. We investigated the effect that the TA adhesive antibiotic had on adhesion and initial growth in urine of Escherichia coli on silicone rubber. The TA antibiotic had reduced adhesion, and inhibited initial growth of the bacteria on the surface. Since adhesion and initial growth on the surface are an essential part of biofilm formation and subsequent infection, we speculate that the TA antibiotic coating might decrease the infection rate associated with indwelling urinary catheter. PMID- 11040436 TI - Some safety aspects of Salmonella vaccines for poultry: in vivo study of the genetic stability of three Salmonella typhimurium live vaccines. AB - Live vaccine strains of Salmonella should be avirulent, immunogenic and genetically stable. Some isolates of three commercially available live vaccine strains of Salmonella typhimurium, sampled during a study on their persistence in a vaccinated flock of chickens, were analyzed for genetic stability using macrorestriction analysis of their genome. Two out of the three vaccine strains showed genetic instabilities. Two of the 51 isolates of Zoosaloral vaccine strain and nine of the 32 analyzed isolates of chi(3985), a genetically modified organism, were variants and showed different macrorestriction profiles. PMID- 11040437 TI - Variation of the mexT gene, a regulator of the MexEF-oprN efflux pump expression in wild-type strains of Pseudomonas aeruginosa. AB - We found three variations of wild-type strains in terms of mexT-mediated regulation of the MexEF-OprN efflux pump, in which overexpression of the pump results in nfxC-type antibiotic resistance in Pseudomonas aeruginosa. Type-I: the mexT gene of the wild-type strain encoded inactive MexT and the nfxC-type mutants derived from this parent had an additional mutation in mexT converting MexT from the inactive to the active form. Type-II: The mexT gene in the wild-type strain had an 8-bp insert producing inactive MexT and the nfxC-type mutants from this parent had a deletion of the 8-bp insert converting inactive MexT to the active form. Type-III: Both the wild-type strain and its nfxC-type derivative produced identical and active MexT. The nfxC mutant from this parent must have an additional mutation. The original nfxC mutant isolated in 1990 might be derived from the Type-I parent strain. PMID- 11040438 TI - Genetic variations among Mycoplasma bovis strains isolated from Danish cattle. AB - The genetic heterogeneity of Mycoplasma bovis strains isolated in Denmark over a 17-year period was investigated. Forty-two field strains isolated from different geographic locations and specimens, including strains from 21 herds involved in two outbreaks of M. bovis-induced mastitis, and the type strain of M. bovis (PG45(T)) were assayed for variations in the BglII and MfeI restriction sites in the chromosomal DNA by using the amplified fragment length polymorphism (AFLP) fingerprinting technique. The obtained genomic fingerprints consisted of 62-68 AFLP fragments in the size range of 50-500 bp. Among the analyzed strains, 18 different AFLP profiles were detected. The similarity between individual fingerprints, calculated by Dice similarity coefficient, ranged from 0.9 to 1.0. Twenty-five strains, including 23 which were isolated during two outbreaks of M. bovis-induced mastitis which occurred 2 years apart, showed indistinguishable AFLP patterns. More genetic diversity was observed among the recent strains. The similarity of the genotypes of the field strains to that of the M. bovis type strain (PG45(T)) was 97.7%. The results of this study have demonstrated a remarkable genomic homogeneity of Danish strains of M. bovis that were probably epidemiologically related and which have remained stable for a considerable length of time. Furthermore, this study has demonstrated that AFLP can be used for genomic fingerprinting and discrimination of M. bovis strains. PMID- 11040439 TI - The life cycles of the temperate lactococcal bacteriophage phiLC3 monitored by a quantitative PCR method. AB - We present here a new and general approach for monitoring the life cycles of temperate bacteriophages which establish lysogeny by inserting their genomes site specifically into the bacterial host chromosome. The method is based on quantitative amplification of specific DNA sites involved in various cut-and-join events during the life cycles of the phages (i.e. the cos, attP, attB, attL and attR sites) with the use of sequence-specific primers. By comparing the amounts of these specific DNA sites at different intervals, we were able to follow the development of the lytic and lysogenic life cycles of the temperate lactococcal bacteriophage phiLC3 after infection of its bacterial host Lactococcus lactis ssp. cremoris IMN-C18. PMID- 11040440 TI - Characterisation of the bovine enteric calici-like virus, Newbury agent 1. AB - The bovine enteric calici-like virus, Newbury agent 1 (NA1) was characterised to determine if it is a member of the Caliciviridae and to establish its antigenic relationship to the established bovine enteric calicivirus Newbury agent 2 (NA2). Solid phase immune electron microscopy (SPIEM) allowed quantification of NA1 virions and identification of faecal samples with optimal virus levels. NA1 particles were 36.6 nm in diameter, had an indefinite surface structure resembling that of human small round structured viruses (SRSVs), and a buoyant density of 1.34 g ml(-1). A single capsid protein of 49.4 kDa was detected by Western blotting in purified NA1 preparations prepared from post-infection but not pre-infection faecal samples and with post- but not pre-infection sera. NA1 was antigenically unrelated to the bovine enteric calicivirus NA2 by SPIEM. These properties were consistent with classification of NA1 within the Caliciviridae but demonstrated heterogeneity in the capsid composition of bovine enteric caliciviruses. PMID- 11040441 TI - Mechanism of oxidative DNA damage induction in a strict anaerobe, Prevotella melaninogenica. AB - We investigated the mechanism of the oxidative DNA damage induction by exposure to O(2) in Prevotella melaninogenica, a strict anaerobe. Flow cytometry with hydroethidine and dichlorofluorescein diacetate showed that O(2) exposure generated O(2)*-) and H(2)O(2). Results of electron spin resonance with alpha-(4 pyridyl-1-oxide)-N-tert-butylnitrone and ethanol showed that O(2) exposure also induced *OH radical generation in P. melaninogenica loaded with FeCl(2) but not in samples without FeCl(2) loading. In P. melaninogenica, O(2) exposure increased 8-hydroxydeoxyguanosine (8OHdG), typical of oxidative DNA damage. Catalase inhibited the increase, but the *OH radical scavengers did not. Phenanthroline, a membrane-permeable Fe and Cu chelator, increased the 8OHdG induction. In FeCl(2) loaded samples, induction of 8OHdG decreased. Addition of H(2)O(2) markedly increased 8OHdG levels. These results indicate that in P. melaninogenica, exposure to O(2) generated and accumulated O(2)* and H(2)O(2), and that a crypto OH radical generated through H(2)O(2) was the active species in the 8OHdG induction. PMID- 11040442 TI - Production of shiga toxin by Escherichia coli measured with reference to the membrane vesicle-associated toxins. AB - Production of Shiga toxin (Stx) 1 and 2 from Stx-producing Escherichia coli (STEC) was measured with reference to the membrane vesicle (MV)-associated toxins. An immunoblot analysis method using specific antibodies for Stx1 and Stx2 was developed for the detection of the extracellular toxins. All 46 STEC isolates, studied including 30 O157 and 16 other O-antigenic isolates, released Stx1 and Stx2 as MV-associated and MV-removed fractions under aerobic and anaerobic conditions. Treatment of vesicles with polymyxin B that disrupted MVs increased the release of Stx1 and Stx2. Therefore, delivery of Stx1 and Stx2 by MVs is a general mechanism in STEC. Stx1 remained within MVs rather than in the MV-removed fraction under an aerobic culture condition. On the other hand, a larger proportion of Stx2 was detected in the MV-removed fraction. The kinetic patterns of the release of the toxins from STEC strains showed that both Stx1 and Stx2 were released into the growth medium during the exponential growth phase. An rpoS-deficient mutation did not have altered levels of extracellular Stx1 and Stx2, supporting the idea that Stx1 and Stx2 are produced during exponential growth phase. PMID- 11040443 TI - Localization of Legionella bacteria within ribosome-studded phagosomes is not restricted to Legionella pneumophila. AB - In this report, we investigate the intracellular fate of selected members of the genus Legionella within the monocytic cell line Mono Mac 6 cells. By means of electron microscopy and immunocytochemistry, we could show that Legionella pneumophila as well as Legionella longbeachae are able to induce ribosome-studded phagosomes which associate with the rough endoplasmic reticulum (RER), whereas Legionella micdadei remains to be located within smooth phagosomes but also shows signs of RER association. In addition, we could demonstrate a remarkable correlation between the phagosome type and the morphological phenotype of intracellular bacteria: within ribosome-studded phagosomes, bacteria generally lacked the outer coat of low electron density whereas bacteria within the smooth phagosomes still possessed this outer coat. The virulence factors responsible for inhibition of phagosome maturation and their distribution within the genus Legionella as well as the biological significance of the morphological difference of bacteria within smooth and ER-associated phagosomes remain to be investigated. PMID- 11040444 TI - The membrane potential of Giardia intestinalis. AB - Giardia intestinalis is a primitive microaerophilic protozoan parasite which colonises the upper intestine of humans. Despite the evolutionary and medical significance of this organism, its physiology is very poorly understood. In this study we have used a novel flow cytometric technique to make quantitative measurements of the electrical potential across the plasma membrane of G. intestinalis trophozoites. In media lacking both K(+) and Na(+), G. intestinalis trophozoites maintained a high negative plasma membrane potential (Psi(m)) of 134+/-3 mV. The Psi(m) was unaffected by the addition of Na(+) to the extracellular medium, whereas the addition of K(+) resulted in a significant membrane depolarisation, consistent with the G. intestinalis trophozoite plasma membrane having a significant (electrophoretic) permeability to K(+). The membrane was also depolarised by the H(+) ionophore m-chlorophenylhydrazone and by the H(+) ATPase inhibitors dicyclohexylcarbodiimide and N-ethylmaleimide. These results are consistent with G. intestinalis trophozoites maintaining a high resting Psi(m), originating at least in part from an electrogenic H(+) pump acting in concert with a K(+) diffusion pathway. PMID- 11040445 TI - Pancreatic carcinoma is characterized by elevated content of hyaluronan and chondroitin sulfate with altered disaccharide composition. AB - The amount and the types of glycosaminoglycans (GAGs) present in human pancreatic carcinoma were examined and compared with those in normal pancreas. Human pancreatic carcinoma contained increased levels (4-fold) of total GAGs. Particularly, this carcinoma is characterized by a 12-fold increase of hyaluronan (HA) and a 22-fold increase in chondroitin sulfate (CS) content. CS in pancreatic carcinoma exhibited an altered disaccharide composition which is associated with marked increase of non-sulfated and 6-sulfated disaccharides. Dermatan sulfate (DS) was also increased (1.5-fold) in carcinoma, whereas heparan sulfate (HS), the major GAG of normal pancreas, becomes the minor GAG in pancreatic carcinoma without significant changes in the content and in molecular size. In all cases, the galactosaminoglycans (GalGAGs, i.e. CS and DS) derived from pancreatic carcinomas were of lower molecular size compared to those from normal pancreas. The results in this study indicate, for the first time, that human pancreatic carcinoma is characterized by highly increased amounts of HA and of a structurally altered CS. PMID- 11040446 TI - A selective inhibitor of p38 MAP kinase, SB202190, induced apoptotic cell death of a lipopolysaccharide-treated macrophage-like cell line, J774.1. AB - A selective p38 MAP kinase (p38 MAPK) inhibitor, SB202190, induced apoptotic cell death of a macrophage-like cell line, J774.1, in the presence of lipopolysaccharide (LPS), as judged by DNA nicks revealed by terminal deoxy transferase (TdT)-mediated dUTP nick end labeling (TUNEL), activation of caspase 3, and subsequent release of lactate dehydrogenase. This cytotoxicity was dependent on both LPS and SB202190, and such inhibitors of the upstream LPS signaling cascade as polymyxin B and TPCK blocked this macrophage cell death. SB202190 suppressed the kinase activity of p38, leading to inhibition of activation of MAPKAPK2 and then the subsequent phosphorylation of hsp27 in LPS treated macrophages both in vitro and in vivo, but an inactive analog of SB202190, SB202474, did not. There was a threshold of the time of addition of SB202190 to LPS-treated macrophages to induce apoptosis, which was before full transmission of p38 activity to a direct downstream kinase, MAPKAPK2. Besides, localization of phosphorylated hsp27 in Golgi area of the LPS-treated macrophages was suppressed by SB202190, while it was not by SB202474. These results suggest that selective inhibition of p38 MAPK activity in LPS-induced MAP kinase cascade leads to apoptosis of macrophages. PMID- 11040447 TI - Self-regulation of the endothelin receptor system in choriocarcinoma cells. AB - The human trophoblast secretes endothelin-1 (ET-1) and expresses ET receptors. The present study tested whether the transformed BeWo, JAR and JEG-3 choriocarcinoma cells: (1) secrete endothelin-1 (ET-1); (2) express both ET-A and ET-B receptor subtypes; and (3) have the potential to allow for autologous regulation of ET-receptor proteins. The cells were cultured for 24/48 h with or without 10% FCS and, in experiments on receptor regulation, with ET-1 (5-20 nM and 10 microM). ET-1 secretion was measured by RIA and receptor levels by immunoblotting. All cell types secreted ET-1 albeit at different levels and sensitivity to FCS. All cell lines expressed both ET-A (JEG-3>BeWo=JAR) and ET-B (JEG-3=JAR>BeWo) receptor subtypes, which could be up- and downregulated depending on ET-1 concentration, culture time and FCS presence. It is concluded that BeWo, JAR and JEG-3 choriocarcinoma cells secrete ET-1 and express both ET-A and ET-B receptor subtypes. The receptor levels can be regulated by ET-1. This provides the molecular basis for an autocrine system with the potential of autologous regulation of yet unidentified ET-1-induced functions. PMID- 11040448 TI - The effect of cysteine on the altered expression of class alpha and mu glutathione S-transferase genes in the rat liver during protein-calorie malnutrition. AB - Protein-calorie malnutrition (PCM) represents a global health problem. The breakdown rate of glutathione S-transferase (GST) subunits determines their differential contents during protein depletion. Hepatic GST expression and the underlying mechanistic basis were investigated in PCM rats. PCM caused no change in rGSTA1/2 subunit. In contrast, rGSTA3/5 subunit was 2.4-fold induced during PCM, while the levels for rGSTM1 and M2 subunits were 30% and 70% suppressed. Increased GSTA3/5 expression was significantly prevented by cysteine or methionine treatment, although such treatment failed to restore the rGSTM2 level. In contrast to differential GST protein expression, PCM caused a 5-10-fold increase in rGSTA2/A3/A5 and M1 mRNAs, whereas rGSTM2 mRNA was 70% decreased. The elevations in rGSTA2/A3/A5 and M1 mRNAs were completely abolished by cysteine or methionine treatment during PCM, although the rGSTM2 mRNA level was not restored. PCM induced oxidative stress in the liver, as evidenced by protein carbonylation. Antioxidant response element (ARE)-binding activity of nuclear extracts from PCM rats was increased, which was immunodepleted with anti-Nrf-1/2 antibodies. Activation of nuclear ARE-binding proteins was inhibited by cysteine. Data showed that hepatic GSTs were differentially expressed during PCM, that certain GST mRNAs were increased with the ARE activation, and that cysteine was active in preventing increases in GST mRNAs and ARE activation. PMID- 11040449 TI - DMSO and glycerol reduce bacterial death induced by expression of truncated N terminal huntingtin with expanded polyglutamine tracts. AB - Huntington's disease (HD) is caused by CAG repeat expansion in exon 1 of a large gene, IT15, possessing 67 exons. Transgenic mice expressing a truncated N terminal peptide of huntingtin with an expanded polyglutamine tract translated only from exon 1 develop symptoms similar to Huntington's disease. In the present study, a bacterial system (Escherichia coli) was used to express truncated peptides of huntingtin translated from exon 1 of the HD gene. Bacterial death was observed after the induction of peptides with expanded polyglutamine tracts, and both sodium dodecyl sulfate (SDS)-soluble peptides and insoluble aggregated material were detected by immunoblotting in the homogenates of such E. coli. E. coli death was partially reduced by the addition of dimethylsulfoxide (DMSO) or glycerol to the medium, with a consequent decrease in aggregated material and an increase in SDS-soluble peptide in the homogenate. These results suggest that DMSO and glycerol may decrease the toxicity of huntingtin with expanded polyglutamine tracts by acting as chemical chaperones. PMID- 11040450 TI - Sera of patients suffering from inflammatory diseases contain group IIA but not group V phospholipase A(2). AB - During recent years, the high phospholipase A(2) (PLA(2)) concentrations at sites of inflammation and in circulation in several life-threatening diseases, such as sepsis, multi-organ dysfunction and acute respiratory distress syndrome, has generally been ascribed to the non-pancreatic group IIA PLA(2). Recently the family of secreted low molecular mass PLA(2) enzymes has rapidly expanded. In some cases, a newly described enzyme appeared to be cross-reactive with antibodies against the group IIA enzyme. For this reason, reports describing the expression of group IIA PLA(2) during inflammatory conditions need to be reevaluated. Here we describe the identification of the PLA(2) activity in sera of acute chest syndrome patients and in sera of trauma victims. In both cases, the PLA(2) activity was identified as group IIA. This classification was based upon cross-reactivity with monoclonal antibodies against group IIA PLA(2) which do not recognize the recombinant human group V enzyme. Moreover, purification of the enzymatic activity from the two sera followed by N-terminal amino acid sequence analyses revealed only the presence of group IIA enzyme. PMID- 11040451 TI - Pharmacokinetics of chronically administered all-trans-retinoyl-beta-glucuronide in mice. AB - After the subcutaneous injection of retinoyl beta-glucuronide (RAG), both RAG and retinoic acid (RA), formed by the hydrolysis of RAG in vivo, achieved peak plasma concentrations within 1-2 h. Thereafter, RA was rapidly cleared from the plasma whereas RAG was eliminated much more slowly. No significant changes were noted in the peak (2 h) plasma levels of RAG for treatment periods up to 56 days (one injection of RAG/day), in the clearance rate of RAG from plasma, or in plasma retinol concentrations. Similarly, no consistent decrease in plasma levels of the RA hydrolysis product was observed. Mice undergoing these long-term chronic treatments with RAG did not show any clinical manifestations of retinoid toxicity. Taken together, our findings that chronic dosing with RAG produces sustained levels of both the parent compound and the RA hydrolysis product, combined with the apparent low toxicity of RAG, suggest that RAG could be a safe and useful alternative to some retinoids which are presently being utilized in the clinic. PMID- 11040452 TI - Inhibition of diethylnitrosamine-induced rat liver chromosomal aberrations and DNA-strand breaks by synergistic supplementation of vanadium and 1alpha,25 dihydroxyvitamin D(3). AB - Vanadium (V) has recently been found to possess potent anti-neoplastic activity in rat hepatocarcinogenesis. Recent studies have suggested that the active metabolite of vitamin D(3), 1alpha, 25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)], can inhibit growth and/or induce differentiation of a variety of cell types. In the present study, attempts have been made to investigate the combination effects on chromosomal aberrations (CAs) and DNA-strand breaks during the early preneoplastic stage of diethylnitrosamine (DEN)-induced rat liver carcinogenesis in male Sprague-Dawley rats. V (0.5 ppm ad libitum) and/or 1,25(OH)(2)D(3) (0.3 microg/0.1 ml propylene glycol per os twice weekly) either alone or in combination were given to DEN-treated and control rats 4 weeks prior to DEN injection. Under these experimental conditions it was observed that, when given in combination, V and 1,25(OH)(2)D(3) offered maximum protection against DEN induced structural aberrations 96 h (66.7%, P<0.05), 15 days (44.9%, P<0.005) and 30 days (63.8%, P<0.001) after DEN injection. Synergistic supplementation of both V and 1, 25(OH)(2)D(3) 4 weeks before DEN injection was found to offer significant (64.1%, P<0.001) protection against generation of single-strand breaks when compared with the DEN control. Thus, the combination effect of V, an essential trace element, and of 1, 25(OH)(2)D(3), a dietary micronutrient, appears beneficial in preventing genetic damage in liver cells upon alkylation induced by DEN. PMID- 11040453 TI - The 18q21 region in colorectal and pancreatic cancer: independent loss of DCC and DPC4 expression. AB - The 18q21 region is frequently altered in gastrointestinal tumors. Three candidate tumor suppressor genes have been identified in it: DCC, Smad4/DPC4 and Smad2; the mechanisms involving their inactivation have not been completely elucidated. In this study, genetic losses at 18q21 and expression of DCC and DPC4 in colorectal (n=12) and pancreatic (n=16) cell lines and in colorectal tissues (n=10) were analyzed. The status of the 18q21 region was assessed using microsatellite analysis and duplex PCR of exonic sequences; expression was analyzed by RT-PCR; mutational analysis of DPC4 cDNA was performed in selected cases. Homozygous losses of microsatellite markers at 18q21 were not observed in colon or pancreas lines; however, a higher proportion of apparent homozygosity than expected was found. DCC and DPC4 transcripts were detected in 11/12 and 12/12 colorectal cancer lines, respectively. In tumors, homozygous losses at 18q21 were detected in three cases, without affecting DCC. All tumors retained DCC and DPC4 mRNA expression. In pancreatic lines, DPC4 was inactivated through homozygous deletion (n=5), intragenic mutation (n=3), and lack of protein (n=2). IN CONCLUSION: (1) microsatellite analysis does not provide adequate information regarding homozygous losses at 18q21; (2) approximately 65% of pancreas cancer lines show inactivation of DPC4; and (3) loss of DCC and DPC4 occur independently. PMID- 11040454 TI - Extracellular ATP and UTP stimulate cartilage proteoglycan and collagen accumulation in bovine articular chondrocyte pellet cultures. AB - Bovine articular chondrocytes were maintained in high density pellet cultures with and without serum and nucleotide triphosphates for different periods of time. Despite half-lives in culture of about 3 h, adenosine triphosphate and uridine triphosphate in the presence of serum increased sulphated glycosaminoglycan and collagen deposition above control levels. In the presence of serum a single dose of uridine triphosphate on the first day of culture was sufficient to induce significant increases in subsequent proteoglycan and collagen deposition. We conclude that both adenine triphosphate and uridine triphosphate are anabolic for articular chondrocytes, and that this effect on the chondrocyte is long-term. PMID- 11040456 TI - Editor's corner. PMID- 11040455 TI - Induction of collagenase-3 (MMP-13) in rheumatoid arthritis synovial fibroblasts. AB - There is a growing body of evidence that implicates matrix metalloproteinases (MMPs) as major players in numerous diseased conditions. The articular cartilage degradation that is characteristic of rheumatoid arthritis (RA) is believed to be mediated by the collagenase subfamily of matrix metalloproteinases. The preference of collagenase-3 (CL-3) for collagen type II makes it a likely candidate in the turnover of articular cartilage and a potential target for drug development. In this study, RA synovial membrane tissue was shown to express CL-3 mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR) and protein by immunohistochemistry. Fibroblasts isolated and cultured from RA synovial membrane tissue were induced to express CL-3 mRNA. CL-3 mRNA was detected after PMA treatment in 16 of the 18 RA synovial membrane fibroblast cell lines established for this study. These fibroblasts also expressed mRNA for collagenase-1 (CL-1, MMP-1), membrane type-1 matrix metalloproteinase, gelatinase A, gelatinase B, stromelysin-1, stromelysin-2, TIMP-1, and TIMP-2. They were further shown to express CL-1 mRNA constitutively and CL-3 mRNA only after stimulation with PMA, IL-1, TGF-beta1, TNF-alpha, or IL-6 with IL-6sR. These fibroblasts also expressed after induction both CL-1 and CL-3 at the protein level as determined by Western blot analyses and immunofluorescence. PMID- 11040457 TI - Controversies in the treatment of primary insomnia. PMID- 11040458 TI - Nocturia in sleep disordered breathing. PMID- 11040459 TI - Factors influencing the determination of arousal thresholds in infants - a review. AB - Objective: To review the major confounding factors that influence the determination of arousal thresholds in infants.Review of confounding factors: The determination of arousal thresholds in infants measures their arousability from sleep. The evaluation is influenced by various conditions. The infant's arousability is decreased by maternal factors, such as exposure to cigarette smoke, alcohol, illegal drugs or medications during gestation or breastfeeding. The levels of arousal thresholds also depend on the age of the infant and on experimental conditions, such as previous sleep deprivation, type of arousal challenges, time of administration of the arousal challenge, sleep stage, body position, room temperature, use of a pacifier, bed sharing, or type of feeding. In addition, spontaneous arousals can occur and modify the infant's response to external arousal challenges.Conclusions: Factors known to modify infants' arousability from sleep should be controlled during studies designed to determine arousal thresholds. PMID- 11040460 TI - Psychophysiological insomnia: combined effects of pharmacotherapy and relaxation based treatments. AB - Objective: To compare treatment outcomes associated with combined pharmacologic and non-pharmacologic treatments for psychophysiological insomnia.Background: Treatments for insomnia have included a variety of pharmacotherapy and cognitive behavioral interventions, although few studies have investigated the combined efficacy of drug and non-drug therapy.Methods: Forty-one patients with primary insomnia were randomly assigned to one of three treatment groups: (i) estazolam + muscle relaxation, (ii) estazolam + guided imagery, and (iii) estazolam + sleep education. After 4 weeks of active treatment, subjects were withdrawn from medication and followed for an additional 6 months.Results: Significant improvements were observed in self-report measures of total sleep time, sleep efficiency, and wakefulness after sleep onset in the combined drug and relaxation groups, compared to a significant improvement in total sleep time only in the educational control group. At follow-up, all three groups showed significant improvements across the major sleep measures. Positive changes were also observed in quality of life measures, including mood state and self- ratings of daytime arousal.Conclusions: These findings provide support for the value of combined pharmacotherapy and relaxation training in the treatment of psychophysiological insomnia. PMID- 11040461 TI - Normal pregnancy, daytime sleeping, snoring and blood pressure. AB - Objective: Investigation of daytime sleepiness, blood pressure changes and presence of sleep disordered breathing, in healthy young women during pregnancy.Methods: Young, healthy pregnant women between 18 and 32 years of age, seen in three different prenatal care clinics, were enlisted in a prospective study divided in two parts: part 1 of the study consisted of completing a standardized questionnaire on past and present sleep disorders. It also included filling out visual analog scales (VAS) for daytime sleepiness and snoring by the subject and bed partner. Blood pressure measurement was performed at 9 AM as per the WHO protocol. Similar data were collected again at the 6-month prenatal visit and at the 3-month post-delivery visit. At the 6-month visit, ambulatory monitoring of nocturnal sleep using a portable six-channel recorder (Edentrace((R))) was performed at home. Part 2 involved a subgroup of subjects that were randomly selected after stratification based on results of VAS and ambulatory monitoring. It included 1 night of nocturnal polysomnography with esophageal manometry and 24 h of ambulatory BP monitoring with portable equipment with cuff inflation every 30 min.Results: Of the 267 women who participated in part 1 of the study, only 128 consented to enroll in part 2, from which 26 were selected to undergo polysomnography. At the 6-week prenatal visit 37.45% of the subjects reported daytime sleepiness of variable severity. At the 6-month visit, this was noted in 52% of the subjects. Bed-partners reported chronic, loud snoring prior to pregnancy in 3.7% of the study population, but this increased to 11.8% at the 6-month visit. Blood pressure (BP) remained below the pathological range, i.e. less than 150/95 mm Hg, during the entire pregnancy. However, ambulatory monitoring indicated that 37 women, including the loud chronic snorers, had some minor SaO(2) drops during sleep and this same group presented the largest increase in BP between the 6th week and the 6th month prenatal visits. Part 2 included 26 women, 13 from the above identified 37 women and 13 from the rest of the group, chosen randomly, age and body mass index (BMI) matched. Polysomnography did identify two abnormal breathing patterns during sleep: (1) esophageal pressure 'crescendos' associated predominantly with stage 1 and 2 NREM sleep, and (2) 'abnormal sustained efforts' seen predominantly with delta sleep. These abnormal breathing patterns were noted during a significantly longer time during sleep. This group of women with the abnormal breathing patterns were not only chronic snorers but also had significantly higher systolic and diastolic BP increases when compared to the 13 other non-snorers. Six out of the 13 snorers were 'non-dippers' at the 24-h BP recording.Conclusion: Abnormal breathing during sleep (that is frequently, but not always, associated with loud, chronic snoring, and may be a consequence of edema induced by hormonal changes associated with pregnancy), can be seen in otherwise healthy young pregnant women. It may contribute to the symptom of daytime sleepiness. The changes in blood pressure noted were of no pathological significance in our population but could be an added risk factor in high-risk pregnancies. PMID- 11040462 TI - Comparison of three oral appliances for treatment of severe obstructive sleep apnea syndrome. AB - Objective: To compare three different oral appliances: a mandibular advancement device (Snoreguard), a tongue retaining device, and a soft palate lift, for treatment of severe obstructive sleep apnea syndrome (OSAS).Background: Oral appliances are therapeutic options for patients with OSAS.Methods: Eight patients with a mean apnea hypopnea index (AHI) of 72.1 (SD+/-39.9) were studied. Polysomnographic measures during each of the treatment nights were compared to baseline.Results: Eight out of 8 patients completed the mandibular advancement device (MAD) night; 5/8 tolerated the tongue retaining device (TRD); only 2/8 could sleep with the soft palate lift (SPL) in place. Improvement using the MAD reached significance: overall AHI (mean+/-SD) decreased from 72.1+/-39.9 at baseline to 35.5+/-39.4 with the appliance in place (P<0.02). There was a non significant increase in slow wave sleep (SWS) from 9.6%+/-8.7 to 14.4%+/-10.5 with the MAD in place. In five responders, the mean AHI decreased from 60.0+/ 36.6 to 9.0+/-4.8; all were subjectively improved, using the MAD at 1 year follow up. However, three non-responders had persistence of AHI>40. With the TRD, AHI decreased from 50.3+/-18.9 at baseline to 43.5+/-32.5 (ns). The SPL was not effective with an AHI at baseline of 52.4+/-8.0, and 47.3+/-31.0 with the device in place (ns), and not well tolerated.Conclusions: A mandibular advancement device is an effective treatment alternative in some patients with severe OSAS. In comparison, the tongue retaining device and the soft palate lift do not achieve satisfactory results. PMID- 11040463 TI - The clinical presentation of childhood partial arousal parasomnias. AB - Objectives: The goal of the current study was to compare the sleep characteristics of children diagnosed with a partial arousal parasomnia to a community sample and further, to compare children diagnosed with sleep terrors to those diagnosed with sleepwalking.Background: Many children experience frightened awakenings, with up to 15% meeting criteria for a parasomnia. Despite this, very little empirical data exists examining parasomnias in childhood.Method: The parents of children (between 2 and 12 years of age) referred to a pediatric sleep disorders clinic completed the Children's Sleep Habits Questionnaire (CSH) on their child. The group meeting criteria for partial parasomnia was then matched with a community sample to identify differences in sleep characteristics between children with parasomnias and a normative sample.Results: Children with parasomnias had higher rates of bedtime resistance, sleep onset delay, night waking, and reduced sleep duration than a matched community sample. Sleepwalkers had more sleep onset problems than children with sleep terrors. Almost one quarter of sleepwalkers between 3 and 12 years of age reported nocturnal enuresis.Conclusions: Children with partial arousal parasomnias do have slightly more disturbed sleep than community controls. PMID- 11040464 TI - The use of citalopram in resistant cataplexy. AB - Background: Cataplexy is a disabling component of the narcolepsy tetrad that is sometimes resistant to standard treatment.Case reports: Three of our patients with narcolepsy, including one who had post-traumatic narcolepsy, suffered from intractable cataplexy with failure of treatment with established drugs due to unacceptable side-effects.Results: We explored the use of citalopram (Celexa), the newest and most specific of the serotonin reuptake inhibitors, and were successful in treating cataplexy without significant side-effects. Stimulant drugs remained necessary for controlling symptoms of excessive drowsiness.Conclusions: Citalopram was effective in relieving the symptoms of resistant cataplexy in out patients. PMID- 11040465 TI - Journal search and commentary. PMID- 11040466 TI - Article reviewed: A simplified method for monitoring respiratory impedance during continuous positive airway pressure. PMID- 11040467 TI - Article reviewed: Reduced striatal dopamine transporters in idiopathic rapid eye movement sleep behaviour disorder: comparison with Parkinson's disease and controls. PMID- 11040468 TI - Article reviewed: Abnormalities in CSF concentrations of ferritin and transferrin in restless legs syndrome. PMID- 11040470 TI - Controversies in sleep medicine. PMID- 11040469 TI - Article reviewed: Association of sleep-disordered breathing, sleep apnea, and hypertension in a large community-based study. PMID- 11040472 TI - WebWatch. PMID- 11040471 TI - Sleep-related violence: does the polysomnogram help establish the diagnosis? AB - With the growing number of legal cases of sleep-related violence coming to public attention, and the development of sleep medicine as an area of expertise, sleep clinicians are being turned to for help in discriminating those violent individuals who sustain a diagnosis of adult parasomnia of the arousal disorder type, (sleep walking, 307.46-0, sleep terrors 307.46-1, or confusional arousals 307.46-2) according to the International Classification of Sleep Disorders (The International Classification of Sleep Disorders: revised: diagnostic and coding manual. Rochester MN, American Sleep Disorders Association, 1997) from those whose episode of violence may have been carried out with full waking consciousness, or as the result of an impairment of judgement due to some psychiatric or neurological disorder. Clearly there is need to refine the diagnostic characteristics to resolve some of the contradictory descriptions of this disorder in the present literature. There is also the question whether the classical overnight polysomnogram is helpful in making this differential diagnosis, and if so, how should it be done to be maximally useful and what other testing is indicated. PMID- 11040473 TI - Amblyopia? PMID- 11040474 TI - Optic nerve hypoplasia and small eyes in presumed amblyopia. AB - PURPOSE: To assess the anatomy of eyes presumed to be amblyopic and their fellow eyes. PARTICIPANTS: Two hundred seventy-five patients diagnosed with amblyopia and 88 healthy or glaucomatous subjects. METHODS: All subjects underwent complete examinations, including cycloplegic refraction, slit lamp examination, ophthalmoscopy, and retinal imaging. Axial lengths were determined on 263 amblyopic and 88 healthy and glaucomatous subjects by ultrasonic biometry. MAIN OUTCOME MEASURES: Optic disc areas were determined by magnification correction of disc images performed with formulas. Dysversion of the optic disc was determined by simultaneous viewing of disc photographs, digitized images of both eyes, or both. RESULTS: The mean disc area of eyes presumed to be amblyopic was 1.72 mm(2) +/- 0. 73 SD and 1.95 mm(2) +/- 0.69 SD for nonamblyopic eyes (P =.0017). The mean disc area for 176 optic discs of glaucomatous and healthy eyes was 2.61 mm(2) +/- 0.95 SD. The mean axial length for eyes in the general population is 23.65 mm +/- 1.35 SD. The healthy and glaucomatous group in this study had a mean axial length of 23.89 mm +/- 1.29 SD. The eyes with poorer vision that were assumed to be amblyopic averaged 22.42 mm +/- 2.01 SD in length, whereas their nonamblyopic fellow eyes averaged 22.83 mm +/- 1.89 SD (P =.022). The differences between eyes in the healthy population and eyes that are presumably amblyopic, as well as the healthy and fellow eyes, are highly significant (P <.0001)(7.0 x 10( 16)). CONCLUSION: Vision impairment in presumed amblyopia is associated with optic nerve hypoplasia with relative microphthalmos, which is more notable in those eyes with poorer vision. PMID- 11040475 TI - Two-muscle surgery for congenital esotropia: rate of reoperation in patients with small versus large angles of deviation. AB - INTRODUCTION: Standard surgical treatment of congenital esotropia (CET) in patients with preoperative angles of deviation measuring 50 PD) CET. METHODS: Medical records of 102 patients who underwent bilateral medial rectus muscle recessions between January 1991 and December 1997 were reviewed. Patients were excluded if neurologic abnormalities or developmental delays were documented before the operation, if major structural abnormalities of the eye were present, or if less than 1-month follow-up after surgery was documented. The remaining 56 patients were assigned to either the larger angle (>50 PD) or smaller angle (< or =50 PD) group, based on the magnitude of their preoperative esotropia. Rates of reoperation for residual CET, for consecutive exotropia or dissociated horizontal deviation, or for dissociated vertical deviation with or without oblique muscle dysfunction were determined for each group. RESULTS: Forty of 56 patients (71%) were assigned to the smaller angle group and 16 of 56 patients (29%) to the larger angle group. In the larger angle group, 4 patients (25%) underwent surgery for residual esotropia. In the smaller angle group, 8 patients (19%) underwent surgery for residual esotropia, 8 (19%) underwent surgery for consecutive exotropia or dissociated horizontal deviation, and 8 (19%) underwent surgery for dissociated vertical deviation or oblique muscle dysfunction. CONCLUSION: The success rate for ocular realignment in patients with CET by using bilateral medial rectus muscle recession did not appear to diminish when applied to deviations greater than 50 PD as compared with smaller angle deviations. Surgery on 3 or 4 horizontal rectus muscles may be unnecessary in the treatment of patients with very large angles of CET. PMID- 11040476 TI - Extraocular muscle responses to orbital cooling (ice test) for ocular myasthenia gravis diagnosis. AB - BACKGROUND: As a result of clinical and laboratory investigations of temperature correlates of myasthenia gravis, orbital cooling (ice test) has been developed as a reliable test for ocular myasthenia diagnosis through blepharoptosis response. The test has not been utilized in a prospective manner for myasthenia diagnosis through extraocular muscle responses. METHODS: Fifteen patients with acquired motility disorders were studied with the use of orbital cooling and other tests for myasthenia gravis. Orbital cooling was performed in a standard fashion for all patients. In 14 of 15 patients, the diagnosis of myasthenia was not established at the time the ice test was performed. Fifteen non-myasthenic patients with acquired motility disorders were also studied with use of the ice test. Temperatures during orbital cooling were measured in the superior cul-de sac of one patient and between the lateral rectus muscle and globe in 3 patients. RESULTS: All patients subsequently proven to have myasthenia gravis by other tests and by response to myasthenia therapy had a positive (diagnostic of myasthenia) response to the ice test. No patient had a false-positive or a paradoxical response to the ice test. No control patient had a positive blepharoptosis or motility response to orbital cooling. Temperature measurements demonstrated significant cooling effects in the superotemporal cul-de-sac and beneath the lateral rectus muscles after orbital cooling for 5 minutes. CONCLUSIONS: Orbital cooling, within certain parameters, can be a useful clinical test for myasthenia diagnosis through motility response, as well as blepharoptosis response. PMID- 11040477 TI - Compliance with outpatient follow-up recommendations for infants at risk for retinopathy of prematurity. AB - OBJECTIVES: We undertook this study to determine how frequently at-risk infants were scheduled for and brought to follow-up appointments for retinopathy of prematurity (ROP) examination after hospital discharge and to identify barriers to follow-up. METHODS: The records of 126 infants with or at risk for ROP at the time of hospital discharge were retrospectively reviewed. Data recorded included the presence or absence of a timely outpatient follow-up appointment, the identity of the person who scheduled the appointment (hospital staff or parents), attendance rate for appointments made, race or ethnicity, and insurance status. RESULTS: Sixty-two of 126 (49%) infants were scheduled for a timely outpatient examination. Sixty-four of 126 (51%) required telephone contact from our office to be scheduled for an appointment. Eight of 21 (38%) African American infants had an appointment scheduled without additional intervention by our office personnel, and 6 of 21 (29%) were brought to an appointment in a timely manner. Twenty-two of 33 (68%) white infants had an appointment scheduled without additional intervention by our office personnel, and 20 of 33 (61%) were brought to an appointment in a timely manner. African American patients were less likely than white patients to be brought to a follow-up appointment (P =.022). Eleven of 15 (73%) patients, whose appointments were scheduled by hospital personnel before discharge, were brought to their follow-up appointment, compared with 39 of 105 (37%) patients, whose parents were requested to schedule their own appointment (P =.008). CONCLUSION: Almost 50% of infants with or at risk for ROP were not scheduled for a timely outpatient follow-up appointment, putting these neonates at risk for ROP-related blindness. Patients whose appointments were scheduled by hospital personnel before discharge were more likely to be brought to a follow-up examination. Extensive utilization of office support staff was required to ascertain the status of infants who did not have appointments scheduled or who were not brought to follow-up appointments. PMID- 11040478 TI - Reduction of congenital nystagmus amplitude with auditory biofeedback. AB - PURPOSE: Treatment options for congenital nystagmus without null position are limited. The purpose of this study was to evaluate the role of auditory biofeedback in controlling congenital nystagmus. METHODS: Ten patients with congenital nystagmus without null position underwent 6 sessions (twice a week for 3 weeks) of auditory biofeedback. Each half-hour session had simultaneous electronystagmographic recording done during the session. RESULTS: The patients could reduce the nystagmus during the treatment sessions. Mean amplitude (degrees) of nystagmus was reduced from 6. 28 +/- 4.94 to 3.05 +/- 2.48 (P =.028) and mean intensity (amplitude x frequency) was reduced from 33.37 +/- 22.84 to 13.35 +/- 7.99 (P =. 0174), but the mean frequency change was not significant, from 5.8 +/- 1.05 to 4.98 +/- 1.35 (P =.148). The mean amplitude and mean intensity decreased by 51% and 60%, respectively. After completion of the session, although a subjective improvement was reported, the patient's binocular visual acuity on Snellen's charts and contrast sensitivity did not show any significant change. Also no sustained benefit was noted because the electronystagmographic recordings reverted to baseline after the auditory stimulus for biofeedback was discontinued. CONCLUSION: Simultaneous electronystagmographic recording shows significant reduction of nystagmus amplitude and intensity because of auditory biofeedback only during the treatment session. The beneficial effect does not persist after the auditory stimulus is discontinued. No objective effect on visual acuity and contrast sensitivity was noted after the therapy. PMID- 11040479 TI - Bilateral lateral rectus recession for consecutive exotropia. AB - INTRODUCTION: Treatment of consecutive exotropia after bilateral medial rectus recessions has been rarely studied. Though several series have reported results of medial rectus advancement, none has described results of lateral rectus recessions. METHODS: We reviewed our results in 31 patients who underwent bilateral lateral rectus recessions for consecutive exotropia after bilateral medial rectus recessions. Mean follow-up was 30 months (range, 1-140 months) after the exotropia repair. RESULTS: At last follow-up, 20 of 31 patients (65%) had deviations of less than or equal to 10 PD. Limitation of adduction was not apparent. DISCUSSION: Cooper's dictum states that lateral rectus recession should be performed instead of medial rectus advancement. Our results suggest that this approach is generally successful. The outcome after exotropia repair in consecutive deviations is comparable to that after repair of primary exotropia. PMID- 11040480 TI - Which ocular and neurologic conditions cause disparate results in visual acuity scores recorded with visually evoked potential and teller acuity cards? AB - PURPOSE: We investigated whether disparity between visually evoked potential (VEP) acuity scores and Teller Acuity Card (TAC) scores varied according to presence of ocular or neurologic conditions. METHODS: Charts from 175 children (mean age, 34.8 months; range, 3 to 158 months) referred for visual acuity testing were examined. All children had been tested with pattern-alternation VEP and TAC and had undergone a complete eye examination. VEP and TAC acuity scores were relative to age-expected acuity scores for each acuity test. The absence and degree of macular abnormality, retinal abnormality, optic nerve hypoplasia, optic nerve atrophy, cortical visual impairment, developmental delay, cerebral palsy, seizures, and nystagmus were noted. Analysis of variance models were used to determine whether differences between VEP and TAC scores varied according to the presence of specific deficits. Logistic regression analysis determined whether degree of specific deficits was associated with a greater chance of inconsistency between VEP and TAC scores (>0.3 log unit difference). RESULTS: Inconsistent scores were found in 48% of children. Developmental delay was associated with relatively poorer TAC than VEP score, and the chance of inconsistency increased with severity of developmental delay. CONCLUSIONS: Diagnosis-dependent variability exists between TAC and VEP scores. Therefore knowledge of the clinical picture is necessary in interpretation of VEP and TAC scores. It is not clear which test is more useful when a disparity exists, either from this or previous studies. When visual acuity is assessed longitudinally in a given child, then consistency in method for acuity assessment is important. PMID- 11040481 TI - Comparison of techniques for detecting visually evoked potential asymmetry in albinism. AB - PURPOSE: We compared techniques for analyzing visually evoked potential (VEP) asymmetry in children with albinism to find one that could be used effectively and efficiently. METHOD: Subjects included 21 child volunteers, ages 10 months to 6 years (control group) and 21 children with albinism, ages 2 months to 6 years (albinism group). Five-channel flash VEP was performed on all subjects. Electrodes were positioned at Oz, O1, O2, O3, and O4 (10/20 system). Data were analyzed by use of techniques previously described. These included inspection of the VEP waveforms, measurement of hemispheric waveform parameters, calculation of an asymmetry index, and use of a bipolar derivation between left and right hemispheric responses (interhemispheric difference potential). In addition, we quantified the interhemispheric difference potential by use of Pearson's correlation coefficient. Measurements of sensitivity and specificity determined the success of the 5 analysis paradigms. The accuracy of each paradigm represented the ability to classify the data according to volunteer or albinism group and is derived from both sensitivity and specificity measures. RESULTS: Measurement of hemispheric differences in VEP waveform parameters was the least sensitive measure method for detecting multichannel VEP asymmetry in albinism. Comparison of left and right eye interhemispheric difference potential increased accuracy to 67%. Nonquantitative inspection of waveform demonstrated an accuracy of 76%. The asymmetry index and Pearson's correlate measure yielded accuracy rates of 79% and 83%, respectively. CONCLUSION: The efficiency and capability of Pearson's correlate measure in quantifying interhemispheric difference potentials to detect albinotic misrouting makes this a useful and practical technique in a pediatric clinic. PMID- 11040482 TI - Superior oblique muscle palsy in a patient with orbital dermoid cyst. AB - We describe the clinical and radiologic findings and surgical outcome of an orbital dermoid cyst causing a superior oblique muscle palsy in a child. Superior oblique muscle palsy in childhood is most often congenital. Less common causes are trauma, vascular lesions, neoplasms, and infections.(1,2) The most common orbital lesions in children are dermoid and epidermoid cysts.(3-5) A dermoid cyst at the region of trochlea is suspected as the cause of superior oblique muscle palsy in our case. This unusual presentation of a dermoid cyst has not been reported previously. PMID- 11040483 TI - Ligneous conjunctivitis: a local manifestation of a systemic disorder? AB - Ligneous conjunctivitis is a descriptive term. It refers to the "woody" consistency of the pseudomembrane that usually forms on the palpebral conjunctiva of those affected. It is rare and probably only one manifestation of a multiorgan, pseudomembranous disease. (1-4) We report a case of ligneous conjunctivitis in which investigation revealed a plasminogen deficiency in the heterozygous range (previously reported only in association with a homozygous plasminogen deficiency). We suggest a strategy for investigating known and new cases of ligneous conjunctivitis and/or pseudomembranous disease. PMID- 11040484 TI - Marcus Gunn jaw winking with trigemino-abducens synkinesis. AB - Congenital ocular aberrant innervation syndromes are a complex group of disorders involving abnormal miswiring of the extraocular muscles. This case report describes a child with both a right Marcus Gunn jaw winking phenomenon and a right trigemino-abducens synkinesis, which has not previously been reported in the literature. Clinically, this child presented with an intermittent elevation of the right eyelid and/or an intermittent right exotropia when opening her mouth while sucking or chewing. This case suggests the primary abnormality in this patient may be abnormal development of the trigeminal nerve resulting in the eyelid abnormalities and strabismus. PMID- 11040485 TI - Orbital abscess masquerading as a rhabdomyosarcoma. AB - Although orbital cellulitis is the most common cause of acute-onset proptosis with inflammatory signs in a child, the clinician should always be alert to the possibility of rhabdomyosarcoma. We describe an unusual presentation of acute onset nonaxial proptosis of the left orbit without sinus disease or systemic toxicity in a 6-year-old boy. Our clinical differential diagnosis included orbital cellulitis, metastatic disease, capillary haemangioma, lymphangioma with cyst, ruptured dermoid cyst, and orbital rhabdomyosarcoma. Only after orbital biopsy and subsequent microbiologic confirmation were obtained was a diagnosis of chronic orbital abscess tenable. Features in our patient included paucity of symptoms and signs of inflammation. This case illustrates the difficulty in differentiating a chronic orbital infection from orbital rhabdomyosarcoma on the basis of clinical, laboratory, and orbital imaging findings. Possible causes of this unusual presentation are discussed. PMID- 11040486 TI - Visual rehabilitation in a child with diffuse choroidal hemangioma by using aggressive amblyopia therapy with low-dose external beam irradiation. AB - Diffuse choroidal hemangioma is a congenital vascular hamartoma often associated with hemangiomatous lesions of the brain, orbit, and periocular skin (nevus flammeus) in the Sturge-Weber syndrome. Visual loss from diffuse choroidal hemangioma may result from chronic serous retinal detachments causing retinal pigment epithelial, photoreceptor or cystoid degeneration, and glaucomatous optic atrophy. Low-dose external beam irradiation has successfully resolved exudative retinal detachment and caused shrinkage of the choroidal hemangioma.(1-3) Visual loss in Sturge-Weber syndrome with diffuse choroidal hemangioma often begins during amblyogenic years. Visual rehabilitation may thus require not only therapeutic intervention to address organic disease but also amblyopia therapy. Though many of the previously reported cases address treatment of the diffuse choroidal hemangioma with laser or radiotherapy, none advocate or emphasize treatment of nonorganic amblyopia associated with this condition. We report the case of a child with Sturge-Weber syndrome and unilateral diffuse submacular choroidal hemangioma who developed an exudative retinal detachment that responded to the combination of low-dose external beam irradiation and aggressive amblyopia therapy. PMID- 11040487 TI - The challenge of pediatric cataract surgery. PMID- 11040488 TI - Rupture of the anterior lens capsule in Alport syndrome. PMID- 11040489 TI - [20 years without smallpox]. AB - It is 20 years since the 33rd World Health Assembly (WHA) declared that "worldwide eradication of smallpox" was achieved. This was the outcome of many years intensive work of the World Health Organization (WHO) and its member countries. In 1958 the WHA adopted the recommendation that WHO should initiate the eradication of smallpox on a worldwide scale. In 1967 the eradication activities in hitherto endemic countries became more intense. Smallpox affected 31 countries and 15 countries recorded from occasional cases. Every year more than 10 million people contracted the disease and two million of them died. A ten year limit for the eradication was set. Gradually smallpox were eradicated in South America, then in Asia and last in Africa where the last case of endemic smallpox was recorded in 1977 in Somalia. WHO ensured international collaboration, close coordination of activities and mobilization of financial, personal and material resources. It ensured also that tested methods were fully applied in the affected countries regardless of their political, religious and cultural differences. In the eradication activities participated hundreds of thousands of local and 700 health professionals from abroad, incl. 20 Czechoslovak epidemiologists. The worldwide costs of eradication amounted to some 300 million dollars, i.e. some 23 million per year. The most important contribution of the eradication of smallpox was in addition to the termination of human suffering, worldwide financial savings estimated to 1-2 billion US dollars per year. These saved personal and financial resources could be used for other important health projects. The eradication of variola was defined as eradication of clinical forms of smallpox not as the final eradication of the variola virus. The importance of laboratories keeping the variola virus increased steeply at the time when clinical cases of smallpox were eradicated. From the beginning of the eighties WHO made an effort to reduce their number to a minimum. Since 1984 strains of variola are officially kept only in two centres collaborating with WHO. The Organization suggested destruction of the kept viruses in 1987, i.e. ten years after the eradication of smallpox. Unfortunately some political and scientific circles did not agree with this intention. Even recommendations to destroy the virus in 1993 and again in 1999 were not accepted. In the nineties fear of bio-terrorism and secret modernization of biological weapons influenced some member countries to change their opinion on the intended destruction of the virus. Despite this in May 1999 the WHA adopted a resolution that the final destruction of all variola strains is the objective of all member countries of WHO and recommended to postpone the destruction of the virus to the year 2002. The reason for postponement is current research of new antiviral preparations and better vaccines. There is again hope that all that will be left of the variola virus will be magnetic signals on computer diskettes. PMID- 11040490 TI - [In Process Citation] PMID- 11040491 TI - [Prevention of recurrence of herpes with poliovaccine]. AB - In the course of 20 years in collaboration with dermatologists and doctors attending adolescents 75 patients were treated suffering for at least four years from 10 relapses of herpes labialis or genitalis every year. The usual treatment mitigated only temporarily their complaints but had no effect on the frequency of relapses. Two thirds of the patients had immunological examinations, in half of them shortage of IgA was found. Only 12 subjects had during comprehensive immunological examinations normal results. After 3-4 years immunomodulation with poliovaccine in rare instances combined with other immune preparations, the condition improved in 67 of 69 patients (97%), in two (3%) it remained unaltered. The procedure involved restricted exposure to tobacco smoke, alcohol, UV rays and sleep deficiency. After a comprehensive immunological examination immunomodulation with levamisol was started. This was followed by the administration of 2-3 drops of poliovaccine, at first every month, when the complaints receded the interval of vaccine administration was prolonged. When the complaints persisted the immunity of the organism was promoted by protection from influenza (vaccination, Remantadin) and other respiratory infections (Isoprinosin, Modimunal) and the general resistance was reinforced (Biostim). The mentioned procedure should be reserved for subjects with herpes persisting for a long time and with frequently relapsing herpes. PMID- 11040492 TI - [Determination of antibodies after immunization with the FSME-Immun vaccine]. AB - During 1997-1999 607 sera were obtained from subjects immunized with vaccine FSME Immun. The presence of antibodies was assessed using EIA TBEV-Ig TEST-Line tests, Clinical Diagnostics, Immunozym FSME of Immuno Co. and the virus neutralizing test. Between the results of tests sufficient correlation was not found to allow valid evaluation of the presence of postvaccination antibodies by several methods. When using the Immunozyme FSME test it was found that on the day of administration of the 4th dose of vaccine 15 subjects of 94 had an antibody titre equal to or < 126 VIEU/ml, on the day of administration of the 5th dose this was the case in 5 of 13 immunized subjects. After administration of the 3rd, 4th and 5th dose adequate, i.e. at least three-year immunity, was acquired, depending on age, by 64.1-97.9% of the immunized subjects. As antibody titres higher than 600 VIEU/ml in different age groups were recorded only in 20.0-61.7% of the subjects, the authors do not consider it desirable in subjects immunized with the Austrian vaccine to prolong the time interval between vaccinations beyond three years. PMID- 11040493 TI - [Skin manifestations of Lyme borreliosis in patients at the 1st Dermato venereologic Clinic of Comenius University Medical School in Bratislava 1996 1998]. AB - 60-70% of Lyme borreliosis is formed by early manifestations, in particular erythema migrans (EM). All vegetative forms of the tick Ixodes ricinus (e.g. full grown tick, nymphae and larvae) transmit the causal organism, the spirochete Borrelia burgdorferi sensu stricto and Borrelia burgdorferi sensu lato (B. garinii, B. afzelii). The objective of the work was to evaluate clinical and epidemiological parameters and to contribute by testing and investigating in greater detail criteria to the early and exact diagnosis of the disease. In a group of 50 patients the time of development of EM was evaluated, the mode of transmission, the incubation period, localization, symptoms of dissemination of B. burgdorferi, the period of treatment and the antibiotic used. The dynamics of the antibody titre against B. burgdorferi (IFA test) were assessed in 21 patients with EM for a period of 1 to 22 months and in 5 patients with acrodermatitis chronic atrophicans (ACA) for 3 to 55 months. In 50% EM developed during the summer months. 66% reported as the vector a tick, 14% insects and 20% did not know. An incubation of 1 and 2 weeks was reported in 34%, the lower extremities were affected in 52%. Manifestations of dissemination were found in 6 patients, in 2 patients EM relapsed. Antibodies (Ab) against B. burgdorferi were present in 38 patients. In 21 patients the dynamics of the antibody titre were followed up for 1 to 22 months and no substantial changes were found. Serum positivity in patients with ACA persisted without change of the titre for several years. In the treatment of EM most frequently doxycycline was administered for two weeks. EM as the early stage of LB is a seasonal disease with a natural focus. If treatment is started in the early stage of infection, antibodies against B. burgdorferi need not develop. There is no correlation between clinical complaints and serological results and the type of treatment. The prognosis of the disease is favourable in the majority of patients. Vaccination offers new possibilities in active protection against Borrelia infection, in particular in endemic areas. For European countries at present a recombined vaccine is being prepared from the surface lipoprotein A (OspA) made from prevalent strains of B. afzelii and B. garinii. PMID- 11040494 TI - [A diagnostic medium for Arcanobacterium haemolyticum and other bacterial species reacting with hemolytic synergism to the equi-factor of Rhodococcus equi]. AB - Colonies of Arcanobacterium haemolyticum on common blood agar can be easily overlooked. Therefore a diagnostic medium was developed, on which A. haemolyticum colonies produce a conspicuous zone of complete hemolysis. The medium under question is blood agar prepared from the Columbia Blood Agar Base and 5% washed sheep erythrocytes sensitised with equi factor (EF) of Rhodococcus equi. Optimally, 10 activity units (AU) of EF per 1 ml were used. EF was titrated on a non-nutrient medium consisting of agar Purified (Difco) and 5% washed sheep erythrocytes sensitized with beta-haemolysin (BL) of Staphylococcus aureus, 10 AU/ml. On the same medium, staphylococcal BL was titrated on the basis of its direct haemolytic effect. Despite the very distinct haemolytic reaction of the control strains evident on the diagnostic medium with EF, A. haemolyticum failed to grow from 2.597 throat swabs examined especially for the particular microbe during the a period of two years. However, A. haemolyticum was isolated on this medium twice from 223 swabs from wounds and skin lesions. The proposed medium makes also the rapid diagnosis of species Corynebacterium ulcerans, Dermatophilus congolensis and Listeria monocytogenes on the basis of haemolytic synergism possible. PMID- 11040495 TI - [Analysis of bacterial meningitis in the Slovak Republic 1991-1998]. AB - The authors present the results of analyses of cases of notified bacterial meningitis in Slovakia in 1991-1998, focused on haemophilic meningitis. They demonstrate the ratio of different sources of infection on the development of disease--Haemophilus influenzae 214 cases (13.6%), Streptococcus pneumoniae 183 (11.6%), Neisseria meningitidis 343 (21.8%), other infectious agents 182 (11.5%) and negative cultivations 652 (41.4%). In children aged 0-5 years 668 cases were notified, i.e. 42.4% of the total number (1,574), in other age groups 906 cases, i.e. 57.6%. The mean notified morbidity of haemophil meningitis in 1991-1998 was 0.5/100,000 population. The highest age specific morbidity was in the group of children aged 0-5 years and varied from 2.5/100,000 in 1991 to 7.7/100,000 in 1998. PMID- 11040496 TI - [The "white plague" in Ostrava]. PMID- 11040497 TI - Probing wavepacket dynamics with femtosecond energy- and angle-resolved photoelectron spectroscopy AB - Several recent studies have demonstrated how well-suited femtosecond time resolved photoelectron spectra are for mapping wavepacket dynamics in molecular systems. Theoretical studies of femtosecond photoelectron spectra which incorporate a robust description of the underlying photoionization dynamics should enhance the utility of such spectra as a probe of wavepackets and of the evolution of electronic structure. This should be particularly true in regions of avoided crossings where the photoionization amplitudes and electronic structure may evolve rapidly with geometry. In this paper we present the results of studies of energy- and angle-resolved femtosecond photoelectron spectra for wavepackets in the diatomic systems, Na2 and NaI. Both cases involve motion through regions of avoided crossings. In Na2, however, wavepacket motion occurs on a single adiabatic potential with an inner and outer well and a barrier between them, while in NaI wavepackets move on the nonadiabatically coupled covalent (NaI) and ionic (Na+I-) potentials. Results of these studies will be used to illustrate the insight into wavepacket dynamics that time-resolved photoelectron spectra provide. For example, in the case of NaI these angle-resolved photoelectron spectra seem to offer some promise for probing real-time dynamics of intramolecular electron transfer occurring in the crossing region of the ionic and covalent states. PMID- 11040498 TI - Time- and frequency-resolved photoionisation of the allyl radical AB - We report picosecond time-resolved pump-probe photoelectron spectra of the allyl radical, C3H5, and the fully deuterated allyl, C3D5, carried out in order to elucidate the primary photophysical processes upon UV excitation. It is shown that the UV bands of allyl decay in a two-step process: the first step is an internal conversion to the lower-lying A-state within 20 ps or less, while the second step is a very fast decay from the A-state to the electronic ground state through a conical intersection. In addition we report the first zero kinetic energy (ZEKE) photoelectron spectrum of allyl, yielding an ionisation energy of 65762 cm-1. PMID- 11040499 TI - Towards disentangling coupled electronic-vibrational dynamics in ultrafast non adiabatic processes AB - Femtosecond time-resolved photoelectron spectroscopy is emerging as a new technique for investigating polyatomic excited state dynamics. Due to the sensitivity of photoelectron spectroscopy to both electronic configurations and vibrational dynamics, it is well suited to the study of non-adiabatic processes such as internal conversion, which often occur on sub-picosecond time scales. We discuss the technical requirements for such experiments, including lasers systems, energy- and angle-resolved photoelectron spectrometers and new detectors for coincidence experiments. We present a few examples of these methods applied to problems in diatomic wavepacket dynamics and ultrafast non-adiabatic processes in polyatomic molecules. PMID- 11040500 TI - Electron solvation dynamics in I-(NH3)n clusters AB - Femtosecond photoelectron spectroscopy (FPES) is used to monitor the dynamics associated with the excitation of the charge-transfer-to-solvent (CTTS) precursor states in I-(NH3)n = 4-15 clusters. The FPE spectra imply that the weakly bound excess electron in the excited state undergoes partial solvation via solvent rearrangement on a time scale of 0.5-2 ps, and this partially solvated state decays by electron emission on a 10-50 ps time scale. Both the extent of solvation and the lifetimes increase gradually with cluster size, in contrast to the more abrupt size-dependent effects previously observed in I-(H2O)n clusters. PMID- 11040501 TI - The dynamics of Rydberg electron wavepackets in NO AB - Rydberg electron wavepackets have been studied in molecular NO for a variety of rotational states of the ion core. Predominantly radial motion of the electron wavepacket is observed which is similar to that previously reported in atomic systems. Interference effects similar to those observed in unperturbed Rydberg series are evident and third and fourth order partial revivals are identified. Most interestingly, when the classical period of electronic motion is close to the classical period of rotation of the molecular ion, the molecular dynamics dominates the electronic dynamics. PMID- 11040502 TI - Quantum beats and Kepler motion in fast competing photoionization and photodissociation processes AB - Electronic and nuclear wavepackets created by coherent excitation of an autoionized and predissociated 'complex' resonance in H2 are studied theoretically using time-dependent multichannel quantum defect theory. The calculations predict that quantum beats between the components of the complex resonance interfere with Rydberg wavepacket (Kepler) motion to yield characteristic 'mixed' flux patterns in the observable time-dependent ionization and dissociation signals. PMID- 11040503 TI - An angle resolved electron-ion recoil vector correlation study of alternate ion dissociation channels in A CF3I+ AB - Electron-ion recoil vector correlations are examined for the ionization and subsequent dissociation of A state CF3I+. The magnitude of the electron and fragment ion recoil vectors permits the energetics of two alternative decays to I+ and CF3+ to be compared, while differences between the angular correlations are interpreted as molecule-frame photoelectron angular distributions which, in the I+ channel, are smeared by molecular rotation between ionization and dissociation. Quantitative estimates of sub-ps I+ decay lifetimes are extracted, indicating very different decay rates for the alternative dissociation channels. Surprisingly, the ka1 and ke photoelectron continua exchange polarization dependence in the I+ channel correlations and vibronic interactions are postulated in explanation. This can also rationalize the non-adiabatic A CF3I+- >I+ decay mechanism and the branching competition between the CF3+ and I+ channels. PMID- 11040504 TI - Photoelectron-fragment ion correlations and fixed-molecule photoelectron angular distributions from velocity imaging coincidence experiments AB - A new technique, based on velocity imaging, has been developed to examine correlations in energy and angle between the several particles formed in dissociative single and double photoionisation at VUV wavelengths. Electrons and positive fragment ions are imaged in coincidence at position-sensitive detectors; the resulting multidimensional data sets contain separable energy distributions, angular distributions and correlations in energy and angle between the particles. In some cases internal or external evidence indicates that pure axial recoil occurs, without molecular rotation. In such cases fixed-molecule photoelectron angular distributions can be extracted and parametrised. Results for H2, N2, NO, CO and O2 are presented. Effects of shape resonances in the ionisation channels are apparent in several cases. PMID- 11040505 TI - Photoelectron spectroscopy of ammonia via a fast predissociative state AB - We report a study on resonance enhanced multiphoton ionization photoelectron spectroscopy (REMPI-PES) involving the fast predissociative A state of ammonia, using nano- and femtosecond lasers. The multiphoton scheme involves (1 + 1), (2 + 2), (2 + 2) + 1 and (2 + 2) + 2 photon processes. We have found a progression of stretching vibrations v1 in the PE spectrum when pumping NH3 A v2 = 0, 1 and 3 as intermediate states. The stretching vibration intensity distributions in the photoelectron spectrum are calculated by using the Chebychev method of the wavepacket propagation. The femtosecond spectrum shows a similar feature to the nanosecond spectrum. However, high laser power also causes band broadening and shifting effect as well as above threshold multiphoton ionization. PMID- 11040506 TI - Pulsed field ionization-photoelectron photoion coincidence spectroscopy with synchrotron radiation: the heat of formation of the C2H5+ ion AB - Pulsed field ionization photoelectron (PFI-PE) spectroscopy combined with ion coincidence detection has been used with multi-bunch synchrotron radiation at the Advance Light Source (ALS) to energy select ions and to measure their breakdown diagram. The resolution for ion state selection achieved with Ar+ (2P3/2, 1/2) employing this PFI-PE-photoion coincidence apparatus is 0.6 meV (full width at half maximum). The production of C2H5+ from C2H5Br was investigated near the dissociative photoionization limit with this pulsed field ionization-threshold photoelectron photoion coincidence (PFI-PEPICO) scheme. Although the PFI-PE spectra of C2H5Br, C2H5I, and benzene show that the production of ions in the Franck-Condon gap regions is quite low, the selectivity for PFI-PE detection and the suppression of prompt electrons is such that we can detect 1 PFI-PE out of 25,000 total electrons s-1. The derived C2H5+ heat of formation from the analysis of the C2H5Br+ breakdown diagram and a critical analysis of other results is 900.5 +/- 2.0 kJ mol-1 at 298 K, or 913.2 +/- 2.0 kJ mol-1 at 0 K. This leads to an ethylene proton affinity at 298 K of 682.0 kJ mol-1. The measured IE of C2H5Br is 10.307 eV. PMID- 11040507 TI - Transition state dynamics of the OH + H2O hydrogen exchange reaction studied by dissociative photodetachment of H3O2-. AB - Dynamics in the transition state region of the bimolecular OH + H2O-->H2O + OH hydrogen exchange reaction have been studied by photoelectron-photofragment coincidence spectroscopy of the H3O2- negative ion and its deuterated analog D3O2 . The data reveal vibrationally resolved product translational energy distributions. The total translational energy distribution shows a vibrational progression indicating excitation of the antisymmetric stretch of the water product. Electronic structure calculations at the QCISD level of theory support this analysis. Examination of the translational energy release between the neutral products reveals a dependence on the product vibrational state. These data should provide a critical test of ab initio potential energy surfaces and dynamics calculations. PMID- 11040508 TI - Mass selective gas phase study of ClO, OClO, ClOO and ClAr by anion-ZEKE photoelectron spectroscopy AB - Anion-ZEKE-photoelectron spectra of ClO-, OClO-, ClOO- and the van der Waals cluster ArCl- have been measured. Refined or new values for the electron affinity of ClO, OClO and ClOO have been found. The peak positions in these spectra are in very good agreement with former ClO- and OClO- anion-photoelectron spectra (K. M. Gilles, M. L. Polak and W. C. Lineberger, J. Chem. Phys., 1992, 96, 8012) and a recent ArCl- anion-ZEKE spectrum (T. Lenzer, I. Yourshaw, M. Furlanetto, G. Reiser and D. Neumark, J. Chem. Phys., 1992, 110, 9578). The higher resolution of our anion-ZEKE-photoelectron spectrum of OClO- led to a refined assignment of the corresponding anion-photoelectron spectrum. In addition, a strong difference in the relative intensities of the vibrational peaks has been found in the anion ZEKE-spectrum of OClO- in comparison with the anion-photoelectron spectrum. For the first time, mass selective spectroscopic information has been obtained for ClOO. The strong similarity to the ArCl- spectrum indicates a weakly bound van der Waals cluster Cl.O2. Binding energies of the anion, neutral ground and neutral excited state could be deduced. These are in good agreement with the electron affinities of Cl and ClOO, but differ from theoretical values (K. A. Peterson and H. J. Werner, J. Chem. Phys., 1992, 96, 8948) by a factor of 4.5 and from thermochemically determined values (J. M. Nicovich, K. D. Kreutter, C. J. Shackelford and P. H. Wine, Chem. Phys. Lett., 1991, 179, 367 and S. Baer, H. Hippler, R. Rahn, M. Siefke, N. Seitzinger and J. Troe, J. Chem. Phys., 1991, 95, 6463) by a factor of 9. PMID- 11040509 TI - PFI-ZEKE photoelectron spectra of the methane cation and the dynamic Jahn-Teller effect AB - High resolution pulsed-field-ionization (PFI) zero-kinetic-energy (ZEKE) photoelectron spectroscopy has been used to record the photoelectron spectra of CH4, CDH3, CD2H2 and CD4. The observed extensive progression of rotationally resolved transitions between 100,800 cm-1 and 104,100 cm-1 reveals for the first time the complex energy level structure of the methane cation. The high resolution enabled the determination of accurate values for the adiabatic ionization potentials of the different isotopomers. Based on a simple one dimensional model for the pseudorotation in the different isotopomers, progress has been made towards the understanding of the Jahn-Teller effect at low energies. The static Jahn-Teller distortion in the ion could be determined directly from the vibrationless photoelectron transition in CD2H2. The analysis of the rotational structure in this spectrum with a rigid rotor model leads to an approximate experimental C2v structure. The dynamics of the other methane isotopomers near the adiabatic ionization potentials is dominated by large amplitude vibrational motions between equivalent structures. The corresponding ground state tunneling motions takes place on a picosecond time scale. PMID- 11040510 TI - Pulsed field ionization-ZEKE spectroscopy of cresoles and their aqueous complexes: internal rotation of methyl group and intermolecular vibrations AB - Pulsed field ionization-ZEKE photoelectron spectroscopy and (1 + 1) R2PI spectroscopy have been applied to the cis- and trans-m-cresol.H2O clusters. The internal rotational structure in the S1 state has been re-assigned, and the potential curve has been determined for the cluster. The PFI-ZEKE spectra of the cis- and trans-isomers show low-frequency bands up to 1000 cm-1 above the adiabatic ionization potential IP0. The low-frequency bands are assigned to the internal rotation of the methyl group, the intermolecular stretching and their combination bands in the m-cresol.H2O cluster cation. Level energies and relative transition intensities are reproduced well by a one-dimensional rotor model with a three-fold axis potential. Potential curves for the internal rotation have been determined for both cis- and trans-isomers of m-cresol.H2O cations. The effect of the cluster formation upon the internal methyl rotation, and the interaction between the methyl rotation and the intermolecular vibration are discussed. PMID- 11040511 TI - Resolved high Rydberg spectroscopy of polyatomic molecules and van der Waals clusters AB - Using sub-Doppler double resonance excitation with Fourier-transform limited laser pulses and pulsed field ionization techniques we were able to resolve individual high n Rydberg states (45 < n < 110) below and above the lowest ionization energy of van der Waals clusters of benzene with the noble gases neon and argon. By choosing various selected J'K' intermediate rotational states we detected and assigned several Rydberg series with nearly vanishing quantum defect. They converge to different limits representing the rotational states in the vibrational states of the cluster cation. Even far above the ionization threshold sharp high-n Rydberg states with a width of 750 MHz are observed converging to intramolecular vibrational states located up to 800 cm-1 above the dissociation threshold of the cluster ion. This points to a slow dissociation rate of the cluster ion in the range of 3 x 10(5) s-1 < k < 5 x 10(8) s-1. In further studies single high Rydberg states of benzonitrile, a polyatomic molecule with an high dipole moment of 4.18 D, were detected in the range from n = 50 to 100. We plan to investigate the influence of the strong anisotropic dipole field of this molecule on the coupling between the high Rydberg electron and the molecular core. PMID- 11040512 TI - Resonance-enhanced multiphoton ionisation photoelectron spectroscopy of the ClO radical: the C 2 sigma- state AB - A (2 + 1) one-colour resonance-enhanced multiphoton ionisation study is carried out on the C 2 sigma- state of the ClO radical in the one-photon energy range 29,500-31,250 cm-1. The ClO radical is produced by one-photon photolysis of ClO2 employing 359.2 nm photons derived from a separate laser. In this way a significant concentration of vibrationally excited ClO in its spin-orbit split X 2 pi omega (omega = 3/2 or 1/2) electronic ground state is produced. In addition to mass-resolved excitation spectra, kinetic-energy resolved photoelectron spectra for the X 3 sigma-(v+)<--C 2 sigma-(v' = 3-5) transitions are measured. These transitions are not completely Frank-Condon diagonal, and indicate a decrease in bond length on removal of the Rydberg electron from the C 2 sigma- state. In addition to an unambiguous assignment of the C 2 sigma- state, valuable information is obtained on the degree of vibrational excitation with which the nascent ClO radical is formed in the photolysis of ClO2. Analysis of the photoelectron spectra is supported by Franck-Condon calculations based on potential energy curves either from experimental spectroscopic parameters, or obtained by theoretical ab initio methods. PMID- 11040513 TI - Stabilization of molecular atoms AB - We demonstrate the possibility of stabilizing the motion of ion-pair states through the use of external electric and magnetic fields. In conjunction with the Coulomb force, these fields can be engineered so as to lead to the creation of outer equilibrium points which can support non-spreading coherent wavepackets and long-lived states. Specific application is made to the H(+)-H- ion pair, recently investigated using threshold ion-pair production spectroscopy (TIPPS). PMID- 11040514 TI - The dynamics of high Rydberg states in the presence of time-dependent inhomogeneous fields AB - This paper presents calculations of the evolution of an optically prepared Rydberg wavepacket in the presence of time-dependent inhomogeneous electric fields and the results have relevance to the stabilization of Rydberg states as appropriate to ZEKE spectroscopy. The field is considered to arise from the combination of an applied field, which may be ramped in time, and the presence of microscopic charges, e.g., a pseudo-random distribution of ions, whose positions may also change with time. The results of the calculations lead to a clearer definition of the conditions under which Rydberg stabilization is achieved, such as in field switching experiments (Baranov et al., Chem. Phys. Lett., 1998, 291, 311), and also confirm the mechanisms by which the randomization of population between blue-shifted and red-shifted Stark states occurs in the presence of micro fields due to ions (Palm et al., Philos. Trans. R. Soc. London, Ser. A, 1997, 355, 1551). The motion of the ions is found to have a significant m-locking effect in the calculations, providing a possible mechanism for the commonly observed long-lifetime tail in the population decay of high-n Rydberg states. PMID- 11040515 TI - Recombination of simple molecular ions studied in storage ring: dissociative recombination of H2O+ AB - Dissociative recombination of vibrationally relaxed H2O+ ions with electrons has been studied in the heavy-ion storage ring CRYRING. Absolute cross-sections have been measured for collision energies between 0 eV and 30 eV. The energy dependence of the cross-section below 0.1 eV is found to be much steeper than the E-1 behaviour associated with the dominance of the direct recombination mechanism. Resonant structures found at 4 eV and 11 eV have been attributed to the electron capture to Rydberg states converging to electronically excited ionic states. Complete branching fractions for all dissociation channels have been measured at a collision energy of 0 eV. The dissociation process is dominated by three-body H + H + O breakup that occurs with a branching ratio of 0.71. PMID- 11040516 TI - Threshold ion-pair production spectroscopy (TIPPS) of H2 and D2 AB - The threshold ion-pair production spectra at the J" = 0 and J" = 1 thresholds of H2 and J" = 0, 1 and 2 thresholds of D2 obtained with single photon excitation are presented. The ion-pair yield spectra of H2 and D2 over these energy ranges demonstrate strong resonant enhancement, parts of which dominate the TIPPS signals, permitting the assignment of the lower states of these resonances. From those thresholds with weak resonant enhancement (the J" = 0 threshold of H2 and the J" = 1 threshold of D2) a very small direct contribution to ion-pair production can be observed. The behaviour of the TIPPS spectra taken with different applied discrimination fields is understood by modeling the field ionization behaviour of a MATI spectrum of H2, containing both the similarly resonantly enhanced v+ = 8 S(0) ionization threshold and the non-resonantly enhanced S(1) ionization threshold. From the H2 J" = 1 and D2 J" = 0 TIPPS spectra the energetic field-free thresholds of the H2 and D2 ion-pair limits were determined to be 139,714.8 +/- 1.0 cm-1 and 140,370.2 cm-1 +/- 1.0 cm-1, respectively. PMID- 11040517 TI - Field induced ion-pair formation from ICl studied by optical triple resonance AB - Multiphoton pathways to the ion-pair states of ICl, at energies close to dissociation, are presented. These very high vibrational levels (v > or = 10(4)) are detected in the I+ and Cl- channels by pulsed field ionisation. Using a variable time delay before field ionisation, ion-pair states up to 50 cm-1 below the dissociation limit are shown to survive for at least 2 microseconds, indicating a stabilisation process analogous to that operating in high Rydberg electronic states. The atomic ion production signal is highly structured both above and below the free ion threshold, indicating the role of doorway states which are coupled to the dense ion-pair manifold near dissociation. This initial coupling appears to be very efficient and competes successfully with radiative decay and further up-pumping. PMID- 11040518 TI - Ion-pair formation observed in a pulsed-field ionization photoelectron spectroscopic study of HF AB - The pulsed-field ionization (PFI) photoelectron (PE) spectrum of HF has been recorded at the chemical dynamics beamline of the advanced light source over the photon energy range 15.9-16.5 eV using a time-of-flight selection scheme at a resolution of 0.6 meV. Rotationally-resolved structure in the HF+(X 2 pi 3/2, 1/2, v+ = 0, 1) band systems are assigned. The spectral appearance of these systems agrees with a previous VUV laser PFI-PE study. Importantly, extensive rotationally-resolved structure between these two vibrational band systems is also observed. This is attributed to ion-pair formation via Rydberg states converging on the v+ = 1 vibrational levels of the HF+(X 2 pi 3/2, 1/2) spin orbit states. These Rydberg states are assigned to the 1 sigma+ part of the nd complexes (sigma, pi, and delta). Ion-pair formation is observed in this study by the detection of F- ions. Some partially rotationally-resolved structure in a previously published threshold photoelectron spectrum is similarly attributed to ion-pair formation (F- detection) through a combination of the v+ = 17 level of the (A 2 sigma+) 3s sigma Rydberg state and the (X 2 pi 3/2, 1/2, v+ = 1) 7d Rydberg states. On the basis of the present study, an accurate experimental value for the dissociation energy of the ground state of HF has been obtained, D0(HF) = 5.8650(5) eV. PMID- 11040519 TI - Investigation of charge localization and charge delocalization in model molecules by multiphoton ionization photoelectron spectroscopy and DFT calculations. AB - In this work we focus on the question to which degree a surplus charge is localized or delocalized in extended molecular systems. Molecules consisting of a flexible tail and the benzene chromophore, such as n-propylbenzene, 2-phenylethyl alcohol and 2-phenylethylamine, are used as model molecules. Their S0-S1 resonance enhanced multiphoton ionization (MPI) spectra containing origin transitions of different conformers appear at similar wavelengths. This shows, that in the neutral the electronic excitation is localized at the benzene chromophore. Geometry differences between the neutral and the cation can be qualitatively derived from intensities of vibrational transitions or the onset behavior in MPI high-resolution photoelectron (MPI-PE) spectra. We identify two possible reasons for structural changes: Charge-dipole interaction and charge delocalization. Whereas both effects can be active for the folded gauche conformers, the charge-dipole interaction is expected to be small for the extended anti conformers and geometry changes are attributed to charge delocalization. Density functional calculations of structures and energies qualitatively confirm the experimental results for all molecules and their conformers. They predict charge delocalization into the end group of below 20% for n-propylbenzene and 2-phenylethyl alcohol. In the case of 2-phenylethylamine the charge is equally shared by the near-isoenergetic charge sites of the benzene chromophore and the amine group. PMID- 11040520 TI - Spectroscopy of excited states of carbon anions above the photodetachment threshold AB - Electronic transitions of C3- and C5- to states lying above the electron affinity of the neutral (EA) have been recorded in the gas phase by laser photodetachment spectroscopy. The excited states are identified by comparison with absorption spectra for the mass-selected ions deposited in neon matrices and with ab initio calculations. The C 2 sigma u (+)-X 2 pi g transition and two higher energy band systems are observed for C3-, corresponding to excitation energies more than 1.5 eV above the EA. In the case of C5- the strongest features, at about 0.6 eV above the EA, are attributed to close lying 2 delta g-X 2 pi u and 2 sigma g(-)-X 2 pi u transitions. The dominant configurations in these states identify them as long lived Feshbach resonances. Lifetimes for these resonances in C3- are estimated to be between 200 fs and 3 ps from the band widths. PMID- 11040521 TI - Spectroscopic observation of vibrational Feshbach resonances in near-threshold photoexcitation of X-.CH3NO2 (X- = I- and Br-) AB - We report the observation of resonance structure in the absorption and X-/NO2- photofragment action spectra of the X-.CH3NO2 (X- = I- and Br-) complexes in the region above the electron detachment threshold. The resonance structure corresponds to peaks which appear at the onsets for vibrational excitation of the -NO2 wag, scissors, and stretch modes of neutral CH3NO2, the modes which most strongly distort upon electron capture into its pi* lowest unoccupied molecular orbital. We attribute the peaks to excitation of vibrational Feshbach resonances of the CH3NO2- transient negative ion, where near-threshold excitation of X .CH3NO2 spectroscopically accesses states of the free electron-CH3NO2 system. PMID- 11040522 TI - CDA position statement on sleep apnea. PMID- 11040523 TI - Sharing ideas. PMID- 11040524 TI - Is the patient always right? PMID- 11040525 TI - The pH of tooth-whitening products. AB - Tooth whitening products may be in contact with intraoral structures for several hours or they may be used daily to whiten the teeth. Consequently, these products should have a relatively neutral pH to minimize potential damage. This study measured the pH of 26 commercially available tooth-whitening products. The pH of the different whitening products ranged from 3.67 (highly acidic) to 11.13 (highly basic). The dentist-supervised home-bleaching products had a mean pH of 6.48 (range 5.66 to 7.35). The over-the-counter whitening products had a mean pH of 8.22 (range 5.09 to 11.13), and the whitening toothpastes had a mean pH of 6.83 (range 4.22 to 8.35). The 3 in-office bleaching products had a pH between 3.67 and 6.53. One-way ANOVA showed that there was a significant difference between the 4 product categories. The most basic pH of all the products tested was 11.13 for the whitening gel of Natural White-Rapid White. The most acidic pH of all products tested was 3.67 for Opalescence Xtra 35% hydrogen peroxide in office bleach. The Least-Squares-Means test showed that the over-the-counter category had a pH significantly different from the other categories (p < 0.05). PMID- 11040526 TI - The marginal-ridge rest seat. AB - Natural canine crowns are preferred as abutments for removable partial dentures because of their root morphology and bony support. However, preparing the rest seat on the lingual surface of a mandibular canine risks perforating the enamel. An alternative rest-seat preparation, on the marginal ridge of the canine crown, conserves tooth structure and provides a rest seat of adequate length and depth to ensure support for a cast framework. To illustrate why this little-discussed rest-seat preparation has endured locally, the rationale and preparation of the marginal-ridge rest seat is discussed from both historical and clinical perspectives. PMID- 11040527 TI - Single-tooth replacement: bridge vs. implant-supported restoration. AB - Options for restoring a single tooth include fixed partial denture, resin-bonded restoration and single-tooth implant. In this paper, we discuss the advantages and disadvantages of these methods and factors that must be considered when choosing between them for the replacement of a single tooth. Although in some cases a fixed partial denture is the most appropriate choice, implants have the advantage of allowing preservation of the integrity of sound teeth adjacent to the edentulous area. PMID- 11040528 TI - Splinting teeth--a review of methodology and clinical case reports. AB - Splinting teeth to each other allows weakened teeth to be supported by neighbouring teeth, although the procedure can make oral hygiene procedures difficult. Several methods for splinting teeth, both extracoronal and intracoronal, as well as the materials commonly used for splinting, are described and illustrated. Two case reports are used to demonstrate the situations in which splinting might be appropriate. PMID- 11040529 TI - The Procera abutment--the fifth generation abutment for dental implants. AB - The Branemark dental implant has undergone progressive development in terms of both the implant body itself and the components connecting the implant to the prosthesis. Many screw and abutment designs have been developed, with various degrees of success. About 15 years ago, CAD (computer-assisted design)-CAM (computer-assisted manufacture) technology was introduced to dentists. More recently CAD-CAM has been used in the manufacture of abutments for implants. This article reviews currently available techniques for creating the Procera custom abutment (Nobel Biocare, Goteborg, Sweden) and outlines appropriate applications for this type of implant. PMID- 11040530 TI - [New diagnostic criteria and classification of diabetes mellitus]. AB - World Health Organisation (WHO) has recently proposed new diagnostic criteria and classification of diabetes mellitus. A major change in diagnostic criteria is lowering of diagnostic fasting plasma glucose level: level of 7.0 mM/L or more in two separate samples is sufficient for the diagnosis. Diagnostic criteria for plasma glucose in 120-min. of oral glucose tolerance test are unchanged. Newly recommended fasting level seems to correlate better with 120-min. value and to be a good marker of increased cardiovascular risk. The new classification describes impaired glucose regulation with two stages: impaired fasting glycaemia (plasma glucose of 6.1-7.0 mM/L) which is a new category and impaired glucose tolerance. Both subcategories are not real clinical entities, but markers of diabetic and cardiovascular risk. Diabetes mellitus, as a clinical entity, is separated in four classes: type 1, type 2, other specific types and gestational diabetes. Gestational diabetes includes any glucose intolerance in pregnancy. The Croatian Board for Diabetes Mellitus recommends acceptance of these criteria and classification for clinical use in the country and suggests that OGTT be performed for metabolic syndrome detection in cases of impaired fasting glycaemia. PMID- 11040531 TI - [The effect of maternal smoking on pregnancy outcome]. AB - The aim of the study was to determine the influence of smoking on the pregnancy outcome. The retrospective study was based on the original database made at the Department of Gynecology and Obstetrics, Medical School, University of Rijeka, in the period between 1987 and 1997. In the mentioned period we analysed 37,417 total singleton births out of which there were 1,685 (4.5%) preterm and 35,732 (95.5%) term births, as well as 1,739 (4.6%) hypotrophic and 35,678 (95.4%) eutrophic newborns. The analysis of smoking during the whole pregnancy on the gestation outcome was performed in 9,895 (26.4%) parturient smokers and 27,522 (73.6%) parturient nonsmokers (control group). The frequency of preterm and/or term delivery, intrauterine fetal growth retardation, Apgar score, birth weight, as well as morbidity and perinatal mortality, were analysed in both groups. The frequency of preterm deliveries was 5.4% (n = 529) in parturient smokers and 4.2% (n = 1,156) in parturient nonsmokers. This difference is statistically significant (p < 0.001). The frequency of hypotrophic newborns born to parturient smokers (n = 631 or 6.4%) was statistically significantly higher (p < 0.001) compared to the frequency of hypotrophic newborns (n = 1,108 or 4.0%) born to parturient smokers. The obtained results point to the importance of chronic smoking during pregnancy as a risk factor in the development of maternal and/or fetal complications. PMID- 11040532 TI - [The effect of implementation of a unit dose drug distribution system on drug consumption]. AB - As an important drug consumer in Dubrava University Hospital, Department of Cardiac Surgery has been chosen for testing a new model of drug distribution system known as unit dose drug distribution system. During the first 39 weeks in 1996--comparative period, drugs were delivered from the Pharmacy to the Department of Cardiac Surgery in traditional way, known as floor stock system. Next 65 weeks, until the end of 1997--pilot study period, drugs were delivered directly from the Pharmacy to the patients, using unit dose drug distribution system. Consumption of drugs was measured every week by statistical unit DDD/100 hospital days (Defined Daily Dose) according to Anatomic-Therapeutic-Chemistry (ATC) classification of drugs. For statistical measurements, beside common arithmetic means, geometric means were used which are less sensitive to extreme values of drug consumption. During comparative period drug consumption was chaotic with great oscillations around mean value, while in pilot study period that process was without great oscillations around lower mean value and did not exceed the limits of process. Drug consumption was completely under control, so it was a predictable process. In the pilot study period total drug consumption was 39% less, while consumption of drugs from group C was 30% less. During comparative period group C makes 34%, while in the pilot study period it makes 38% of the total drug consumption. This model of drug distribution in hospital leads to a rationalization of drug consumption and great savings. The pharmacist physician interactive role began to emerge as a direct result of these changes in the drug distribution system. Hospital pharmacist has become a visible member of health care team who is responsible for Quality of all medication-related activities and thus has taken opportunity for clinical pharmacy practice. PMID- 11040533 TI - [Fundamentals of quality control systems in medical-biochemical laboratories--the role of marketing]. AB - The basic criterion for the overall quality system in medical biochemistry laboratories concerning equipment, premises and laboratory staff in primary health care (PHC) (Regulations on quality systems and good laboratory practice of the Croatian Medical Biochemists Chamber, 1995, Regulations on categorization of medical biochemistry laboratories of the Croatian Medical Biochemists Chamber, 1996, EC4: Essential criteria for quality systems in medical laboratories. Eur J Clin Chem Clin Biochem 1997 in medical biochemical laboratories included in the First Croatia health project, Primary health care subproject, has been met by the marketing approach to the project. The equipment ensuring implementation of the complete laboratory program (NN/96), more accurate and precise analytical procedures, and higher reliability of laboratory test results compared with previous equipment, has been purchased by an international tender. Uniform technology and methods of analysis have ensured high standards of good laboratory services, yielding test results than can be transferred from primary to secondary health care level. The new equipment has improved organization between central and detached medical biochemistry laboratory units, while the high quality requirement has led to improvement in the staff structure, e.g., medical biochemists have been employed in laboratories that had previously worked without such a professional. Equipment renewal has been accompanied by proper education for all levels of PHC professionals. PMID- 11040534 TI - [Atrio-biventricular electrostimulation in the treatment of congestive heart failure. Case report]. AB - Over the past few years, indications for permanent cardiac pacing have been broadened. Accordingly, American Heart Association and American College of Cardiology included dilated cardiomyopathy, hypertrophic obstructive cardiomyopathy and pacing for prevention of atrial fibrillation into indications for permanent cardiac pacing. Studies have described favourable effect of dual chamber cardiac pacing in congestive heart failure in dilated cardiomyopathy, regardless of etiology. In the past two years, even more beneficial effect was associated with multisite, biventricular cardiac pacing. On the basis of the reported results, a multisite pacemaker InSync was implanted to a patient with dilated cardiomyopathy (NYHA class IV), who was also on the list for heart transplantation, and who fulfilled other criteria for implantation of multisite pacemaker. During the eleven-month follow-up, functional improvement, better 6 minute walking test and enhanced quality of life of the patient were observed, which is in accordance with the literature data. PMID- 11040535 TI - [Familial adenomatous colonic polyposis]. AB - We described two patients (brother and sister) with familial adenomatous polyposis of the colon. It is an inherited disease with autosomal dominant pattern of inheritance. The incidence is 1:8.000, with usual onset of polyps development late in the first decade of life or during adolescence, and malignant alteration up to the fourth decade of life. APC gene located on long arm of chromosome 5 is responsible for occurrence of the disease that presents with onset of multiple adenomatous polyps in the colon (from some of them to 1000). The treatment includes chemoprevention by sulindac or aspirin that prevents or reverse process of carcinogenesis. Surgical approach is preventive colectomy up to 20 (25) years of life. APC gene mutation (deletion at codon 1309-1311) was proven by DNA analysis from blood and polyp in both patients. There was no evidence of mutations of genes p53 and K-ras. Preventive colectomy is planned as soon as possible. PMID- 11040536 TI - [Anastomosis of the colon using the Valtrac biofragmentable ring]. AB - Since January 1995, Valtrac biofragmentary ring for compressive anastomosis of the colon has been employed in Croatia. This study comprises experience in four Croatian hospitals in which 244 anastomoses of the colon with Valtrac biofragmentary ring were performed in the four-year period (1995-1998). The average age of the patients was 64.7 years. One hundred and eighty-seven of them (76.64%) were operated for colon cancer, 15 (6.1%) for Crohn's disease, eight (3.3%) for rectal cancer, and 34 for other diseases. One hundred and nineteen anastomoses were made between small intestine and colon, and 125 end-to-end of the colon. Valtrac BAR 28 mm in diameter was most frequently used (41.8%), and 34 mm in diameter least often (8.6%). Sixteen patients had complications related to anastomosis. One had obstruction which required repeated surgery, and 15 (6.15%) dehiscence of the anastomosis, of which 12 were reoperated. In the postoperative course six patients (2.46%) died due to the sequelae of dehiscence. Compressive anastomoses with Valtrac biofragmentary ring should be considered equal to manual and stapler methods when choosing the technique of colon anastomosis. PMID- 11040537 TI - [Herpes simplex viruses: biological characteristics, immunopathogenesis, diagnosis and treatment]. AB - Herpes simplex virus (HSV) is the most common cause of sporadic encephalitis in developed countries. Herpes simplex virus type 1 (HSV-1) causes oropharyngeal infections, keratoconjunctivitis and infections of the central nervous system, while herpes simplex virus type 2 (HSV-2) in the immunocompetent most frequently causes genital infections. HSV-1 primary infection usually occurs in the early childhood but is also possible at adolescent age. HSV-2 primary infection is usually postponed till the adult age and coincides with the sexual activity. Common characteristics of these two viruses are a relatively rapid reproductive cycle, an efficient elimination of the infected cells, and the ability of causing a latent infection in the sensory ganglia. Since nowadays there is a specific therapy, the prognoses of severe HSV infections are much better. However, it is necessary that the antiviral therapy be applied shortly after the first symptoms of the disease have appeared. Therefore, the application of a rapid and safe method for detection of HSV from clinical materials is the first step in the treatment of severe and lethal infections like meningoencephalitis. In that light, the method called polymerase chain reaction (PCR) represents a new in vitro technique of DNA replication which enables exponential replication of a well defined DNA fragment. The advantages of this diagnostic method are its rapidity and sensitivity, and it does not require live cells for virus detection. PMID- 11040538 TI - [Adequacy of dialysis: how important is the duration of dialysis?]. PMID- 11040539 TI - [Clinical observations on the nature of early caries. A study with scanning electron microscopy]. AB - BACKGROUND: Early caries lesions present a greater porosity in the deep layers than in the superficial layer of the enamel. The "white spot" lesion is a dynamic lesion which is subjected to the changes occurring in the oral cavity; it can regress, stabilize or progress. The aim of this study is to show if there are considerable morphological differences in the superficial ultrastructural aspect of early caries lesions, active or stabilized, in persons of different age. METHODS: To carry out this SEM observation, 15 patients aged from 12 to 50 presenting early caries lesions have been selected. Some of them had been recently subjected to orthodontic treatment, others several years ago. Five of these patients have been called back to examine again the demineralized area by microscope in the very same place of the first observation. The replica technique was used utilizing a silicon-based impression material, an epoxy resin for replicas casting, little steel nets to fix the silicon material. RESULTS: At the level of white spot lesions it is evident a tissural structure scattered with small overlappings and micropits with a diameter of 0.5-2 mu. The patients who removed orthodontic bands more than 10 years before show a shining white aspect, corresponding to the nearly complete loss of enamel perikymata with well defined abrasions, but without any micropits at SEM examination. CONCLUSIONS: The replica technique has shown to be useful to record the defects of active and stabilized caries lesions. However this technique can't be utilized as a diagnostic instrument because it would be too expensive, but it is certainly a valid support for in vivo research. PMID- 11040540 TI - [In vitro study of periodontal ligament cells]. AB - BACKGROUND: The aim of this research is to outline a procedure able to promote specific cellular differentiation and proliferation with consequent periodontal regeneration. To achieve this goal, use was made of various compounds supposed to have the capacity of aiding periodontal regeneration. METHODS: The cells utilised for this study were obtained from explants of human periodontal ligaments. Their proliferation and differentiation capacity was examined in the presence of: coral granules (350, 500 mu), collagene type 1, growth factors (Platelet derived growth factor, PDGF and Transforming growth factor beta 1, TGF beta 1), both on their own and in different combination with one another. The differentiation activity was evaluated by ultrastructural morphological method (Transmission electron microscope-TEM) and by spectrophotometric investigation of the alkaline phosphatasis (ALP). RESULTS: The data show that the coral granules and among the growth factors used only TGF beta 1 stimulate the differentiation activity of the periodontal ligament cells valued on the basis of their capacity of producing ALP. These data are supported by the observation with TEM. CONCLUSIONS: From these results it is suggested that there may be therapeutic efficiency in the periodontal field of substances promoting cellular proliferation and differentiation. PMID- 11040541 TI - [Mercury determination in human amniotic fluid]. AB - BACKGROUND: One of the components of Ag amalgam is mercury which, owing to its organic derivatives, can pass into the organs and biological fluids. One particularly interesting but worrying aspect of this transition is the possibility that mercury may pass through the placental barrier and reach the fetus. The aim of this study was to evaluate the concentration of total mercury in human amniotic fluid and compare it with the number and occlusal extension of fillings using Ag amalgam. METHODS: A group of 56 pregnant women were selected due to undergo amniocentesis. A dental check-up was carried out in each patient to identify the number and extension of amalgam fillings. Mercury levels in the amniotic liquid were assayed using a spectrophotometer with atomic absorption and a FIAS-amalgam technique. RESULTS: Mercury concentrations in the samples examined ranged from a minimum of 0.00 ng/ml to a maximum of 2.55 ng/ml, mean 0.44 +/- 0.53 ng/ml. The correlations between the variables examined were evaluated by calculating the coefficient of linear regression. No direct relationship was found with mercury levels. The data obtained were used to construct a model of logistic regression showing scant statistical significance (p = 0.05) between the number of fillings and mercury levels, whereas the occlusal extension of dental repairs was significantly correlated with metal concentrations (p < 0.05). CONCLUSIONS: The authors recommend that silver amalgam should be used with considerable caution during pregnancy. PMID- 11040542 TI - [ELISA analysis of salivary cotinine in smokers]. AB - BACKGROUND: The presence in saliva of cotinine, the main and inactive metabolite of nicotine, reflects the extent of systemic distribution of nicotine and explains the increased susceptibility to periodontal disease in smokers. The aim of this study was to investigate the comparative amount of cotinine in the saliva of habitual cigarette smokers, non-smokers and passive smokers. METHODS: Saliva sample were obtained from 14 cigarette smokers and 13 non-smokers (8 passive smokers), all without periodontal disease, and analyzed by Microplate EIA (a variation of ELISA based on cross-reactivity of cotinine with anti-cotinine antibody revealed by absorbance in spectrophotometry) to determine the presence and the amount of cotinine. RESULTS: Cotinine was detected in the saliva of smokers with a mean of 92.3 +/- 4.15 ng/ml and, unexpectedly, there was evidence of cotinine also in the saliva of non-smokers (mean 5.4 +/- 1.22 ng/ml), particularly, in passive-smokers (mean 12.9 +/- 6.67 ng/ml). CONCLUSIONS: The salivary concentration of cotinine can be used to estimate nicotine intake and its possible role in the pathogenesis of periodontal disease also in passive smokers. PMID- 11040543 TI - [Burning mouth syndrome. Clinical experience with 75 patients]. AB - The burning mouth syndrome (BMS) is a very common disorder frequently seen in practical dentistry. It is a particular condition with a complex of strange burning sensation localized in the oral mucosa which is clinically normal. It is very important for the dentist to recognize and classify this particular syndrome in order to exclude other factors which can cause the same symptoms. On the basis of personal experience on 75 patients, the complex management of this particular group of patients has been evaluated to make a right diagnosis and a correct treatment. PMID- 11040544 TI - [Papilloma virus. Review of the literature. Note II. Diagnosis and treatment]. AB - Viral infections due to papilloma virus, are not always easy, for the oral clinician, to diagnose. In this paper the diagnostic procedures to detect infections due to papilloma virus and the available therapies to avoid possible recurrences are pointed out. PMID- 11040545 TI - [Behcet's disease: recent findings. Review of the literature]. AB - In this work the author reviews the clinical literature and the pathogenetic hypotheses, with particular attention to the correlations of the HLA, for Behcet disease. Behcet's syndrome is a multisystem disorder presenting with recurrent oral and/or genital ulcerations, chronic relapsing uveitis that may cause blindness, and neurologic impairments. Although it has a worldwide distribution, the Behcet's disease is rare in the Americas and Europe and is more prevalent in Turkey and the Middle and Far East. It affects mainly young adults, with men having more severe disease than women. Behcet syndrome is often diagnosed in late age for the lack of a correct diagnostic protocol and for the different symptoms that can be present. The need to follow the criteria made by the International Group of study on the disease of Behcet is underlined. PMID- 11040546 TI - [Gorlin-Gotz syndrome in 3 patients from the same family monitored from 1993 to 1999]. AB - The authors describe the clinical symptoms in three subjects belonging to the same family affected by basal cell nevus syndrome. They emphasise that the typical triad of the syndrome (basal-cell nevus, maxillary keratocysts and skeletal anomalies) was present in the subjects examined. The skeletal anomalies were very evident in the eldest daughter, negligible in the younger daughter and absent in the mother. Only two cysts closely related to erupting teeth were removed in the younger daughter between 1992 and 1999, whereas the basal-cell nevi were clearly prevalent compared to other pathological manifestations. Moreover, the authors observe that the cysts increased in volume much more rapidly in the daughters than in the mother: having identified the histological components of the cysts, this phenomenon might be linked to the age of the subjects examined. PMID- 11040547 TI - The use of complementary and alternative therapies. AB - The National Center for Complementary and Alternative Medicine (NCCAM) was created by Congress in 1998 as part of the National Institutes of Health. As interest in alternative and complementary therapies among healthcare providers and consumers has increased in recent years, the NCCAM has provided research funding to determine the efficacy of various types of unconventional treatments. The Center also provides research training and acts as a clearing-house for information dissemination to practitioners and the general public. Inclusion and exclusion criteria for alternative and complementary medicine as defined by the NCCAM are provided. Interest in these non-traditional treatments will likely continue to increase over the next few years. The NCCAM is playing a vital role as it provides avenues to determine how these therapies can lead to enhanced quality of life for individuals as we enter the new millennium. PMID- 11040548 TI - Healing practices: trends, challenges, and opportunities for nurses in acute and critical care. AB - Florence Nightingale espoused a very holistic orientation in nursing. It was her belief that nursing should put the patient in the best possible condition for nature to restore or preserve health, to prevent or cure disease or injury. In other words, nurses should create environments in which healing can happen. Complementary therapies and healing practices offer nurses and patients an expanded array of options to improve health and healing. This article provides an overview of complementary therapies and healing practices and offers suggestions on how to incorporate these therapies when caring for critically ill patients. PMID- 11040549 TI - The self as healer: reflections from a nurse's journey. AB - Complementary and alternative medicine is rapidly making its way into the mainstream through the addition of a variety of therapies in existing settings. Yet, in national surveys, the public seems interested not only in alternative therapies, but also in a more holistic form of healthcare. Nurses have a critical role to play in creating such a healthcare system. Recovering their own identity as healers is a first step in that direction. This article explores the topic of the self as healer through four key questions: what is healing, who or what is the locus of healing, how can nurses facilitate healing, and how does one become a healer? PMID- 11040550 TI - Use of presence in the critical care unit. AB - Nurses have used the intervention of presence for centuries, but only recently has attention been given to defining and describing this intervention that conveys much of the caring aspect of nursing. Presence is more than a nurse's being with a patient physically. Researchers have found that patients recognize and value nurses who are present with their whole beings and are attuned to patients' needs and concerns. When critical care nurses use the intervention of presence, findings have shown that they make a connection with the patient that can lead to earlier identification of patients' problems. Further, critical care nurses can use presence in interactions with patients to avoid the perception by patients and their families that the nurse is emotionally distant or is there just to do a job. By incorporating presence as an integral part of all patient interactions, critical care nurses have the privilege of transforming a technical, potentially impersonal setting into a humane, healing place. PMID- 11040551 TI - Creating a healing environment for elders. AB - The number of elderly, both in society at large and in the critical care population, is increasing at an unprecedented rate. Critical care nurses must address how best to provide care to these elders. The authors focus on physiologic, cognitive, and psychosocial characteristics of the elderly that place them at risk for complications during their stay in critical care. The critical care environment also contributes to complications such as sleep deprivation, sensory deprivation or overload, painful procedures, and decreased social support. The critical care environment may also be a factor in facilitating delirium, common in critically ill elders. Critical care nurses can proactively help to create a healing environment for these elders by facilitating sleep, implementing strategies to reduce delirium, preventing or minimizing painful experiences, and liberalizing family visitations. PMID- 11040552 TI - Awareness: the heart of cultural competence. AB - Cultural competency in critical care is providing care to patients and their families that is compatible with their values and the traditions of their faiths. This requires awareness of one's own values and those of the healthcare system. The nurse must also become aware of the cultural and spiritual values of patients and families. Although knowledge of many cultures is impossible, willingness to learn about, respect, and work with persons from different backgrounds is critical to providing culturally competent care. This article discusses elements essential for increasing cultural competency. PMID- 11040553 TI - Nurses pray: use of prayer and spirituality as a complementary therapy in the intensive care setting. AB - The power that prayer and spirituality exerts on healing cannot be underestimated. Body, mind, and spirit are connected to each other. Although patients in hospitals may have the best medical and nursing care available, many seek alternative or complementary therapies. One adjunctive therapy that has grown in popularity recently is the incorporation of prayer and spirituality into the traditional approaches used with acute and critically ill patients. Spirituality is returning to healthcare because many patients believe in it and seek it as part of their treatment. Although spirituality is only one of the many types of alternative and complementary therapies available to patients, it can be powerful approach to their care. This article explores the use of spirituality with a special focus on prayer. PMID- 11040554 TI - Strategies for implementing a guided imagery program to enhance patient experience. AB - The patient in acute care settings can have severe emotional and physical stresses that are also experienced by family and significant others. The experience in a hospital has often been described as overwhelming; it can evoke feelings of fear, anger, helplessness, and isolation. Guided imagery, one of the most well-studied complementary therapies, is used increasingly to improve patients' experiences and healthcare outcomes. More and more, patients are relying on the use of guided imagery to provide a significant source of strength, support, and courage as they prepare for a procedure or manage the stresses of a hospital stay. This article provides a brief review of the research base for guided imagery and broad indications for its use. It describes key elements of the therapy and outlines steps to implement a program of guided imagery that can be used in variety of settings. PMID- 11040555 TI - Effects of massage in acute and critical care. AB - This is a discussion of the results of a systematic review of 22 articles examining the effect of massage on relaxation, comfort, and sleep. The most consistent effect of massage was reduction in anxiety. Eight of 10 original research studies reported that massage significantly decreased anxiety or perception of tension. Seven of 10 studies found that massage produced physiologic relaxation, as indicated by significant changes in the expected direction in one or more physiologic indicators. In the three studies in which the effect of massage on discomfort was investigated, it was found to be effective in reducing pain. In only three studies was the effect of massage on sleep examined. The methods for measuring sleep were unclear in two of the studies, and results were inconclusive in the other. Further research is needed to investigate the effect of massage on discomfort and promoting sleep. PMID- 11040556 TI - Meridian therapy: current research and implications for critical care. AB - The National Institutes of Health recently recommended further research on the efficacy of acupuncture and allocated Federal funds to stimulate clinical studies. Their decision was based on a growing body of successful research outcomes in which acupuncture was used in the treatment of acute and chronic pain, nausea, circulatory functions, and mood-related behavioral disorders. Despite a burgeoning body of clinical research, the ability to generalize findings has been affected by design flaws, sample type and size, and multiple methods of acupuncture site stimulation. Complicating the communication of findings is use of the term acupuncture when other strategies are used to stimulate the point, such as electrical probes and low-intensity laser. A more appropriate term for acupuncture is meridian therapy, because it encompasses all methods used to treat an acupoint. The purpose of this article is to define meridian therapy with a focus on current clinical perspectives, to review research outcomes in areas important to the care of critically ill patients, to identify issues related to the application of meridian therapy in the clinical arena, and to elaborate practitioner preparation and licensure requirements. PMID- 11040557 TI - Healing touch: applications in the acute care setting. AB - Nursing has been dedicated throughout its history to addressing the physical, psychologic, and spiritual aspects of the patient that influence the healing process. Current nursing practice in acute care is focused increasingly on monitoring equipment, giving medications, and administering medical treatments in a fast-paced environment that affords few opportunities for the deeper human connectedness between the nurse and the one who is ill and suffering. Healing touch (HT) is an energy-based complementary therapy fostering that nurse-patient connection. Nurses are beginning to use HT with their patients to assist in easing pain and anxiety, promote relaxation, accelerate wound healing, diminish depression, and increase a patient's sense of well-being. This article reports a conceptual framework for use of HT in acute care settings, describes specific HT techniques, and reviews numerous studies that have reported positive outcomes of HT as a noninvasive complementary therapy. PMID- 11040558 TI - Humor in critical care: no joke. AB - As the climate of healthcare continues to change, critical care nurses are under increasing stress, which in turn puts them at risk for stress-related illnesses. Nurses can benefit from having a repertoire of healthy coping mechanisms, and one of the means available to them is humor. When used appropriately, humor can have positive psychologic, communication, and social benefits, as well as positive physiologic effects. Without considering the elements of bond, environment, and timing, however, humor can come across as offensive and hurtful. This article explores some of the precursors of stress for nurses, the use of humor to combat stress, how to keep humor constructive, and the importance of being active rather than passive in producing humor. PMID- 11040559 TI - Music therapy as a nursing intervention for patients supported by mechanical ventilation. AB - Music therapy is a nonpharmacologic nursing intervention that can be used as a complementary adjunct in the care of patients supported by mechanical ventilation. This article details the theoretical basis of music therapy for relaxation and anxiety reduction, highlights the research testing the intervention in such patients, and discusses areas of needed research to extend further the implementation of music therapy in critical care nursing practice in an effort to promote a healing environment for patients. PMID- 11040560 TI - Animal-assisted therapy: the human-animal bond. AB - Advanced practice nurses are met with the ongoing challenge of using interventions and practices that are evidence based in the care of their patients. Such practices include traditional as well as complementary and alternative therapies. Animal-assisted therapy is an alternative therapeutic modality that can be used to promote quality of life and positive health benefits. This article reviews the theoretical and scientific basis for the use of animal-assisted therapy in patient care. A pilot study in which the effect of fish aquarium animal-assisted therapy on patients' stress levels was examined is summarized. PMID- 11040561 TI - Real-time, problem-based learning at Hartford Hospital. PMID- 11040562 TI - Prevention of workplace violence: the issue is quality of caring--2. PMID- 11040563 TI - A primer on managed care contracting. PMID- 11040564 TI - The year 2000 problem: ensuring the continuity of service. PMID- 11040565 TI - Reengineering nursing information: technology across a health care system. PMID- 11040566 TI - Why benchmarking alone doesn't make the grade. PMID- 11040567 TI - The art of reckoning. PMID- 11040568 TI - Solving the transportation problem with mobile robots. PMID- 11040569 TI - Mainlining the jocular vein: humor can ease your ailments. PMID- 11040570 TI - Overcoming patient communication logjams: telephony reporting systems. PMID- 11040571 TI - Small panel discussion yields big results. PMID- 11040572 TI - Building clinical platforms. The next generation of redesign. PMID- 11040573 TI - Central Australian Nurse Management Model (CAN Model): a strategic approach to the recruitment and retention of remote-area nurses. AB - This paper introduces a new strategic approach, the Central Australian Nurse Management Model (CAN Model), to manage remote area nursing services. Central Australia is home to approximately 45,000 people, of whom 30% are Aborigines with a health status that is markedly lower than the rest of the population. While the Federal, State and Territory governments have policies in place to address health inequities, improvement has been hindered by the difficulty in recruiting and retaining suitable nursing staff in remote areas. Implementation of the three key initiatives that comprise the CAN Model has succeeded in attracting, stabilising and skilling a remote area nursing workforce, fundamental to achieving better health outcomes in Aboriginal populations. PMID- 11040574 TI - Thinking about a mobile health unit to deliver services? Things to consider before buying. AB - Purchasing a mobile unit to deliver health-care services can be an expensive undertaking for anyone interested in pursuing this option. Yet, little information is found in the literature on planning or designing such vehicles. A set of guidelines could help administrators to make better decisions regarding this approach for delivering healthcare. This article focuses on mobile health units (MHU). It provides a synthesis of the literature in addition to information from written and oral correspondence with the chief executive officers (CEO) of firms that manufacture MHU. On-site visits to agencies using an MHU were made by one of the authors (DM) to glean their perspective. The combined sources led to the development of guidelines and checklists that can assist administrators in planning the function, design and operation of an MHU to deliver health-care services to remote rural sites. PMID- 11040575 TI - Alternative Curricular Options in Rural Networks (ACORNS): impact of early rural clinical exposure in the University of West Australia medical course. AB - The aim of this study was to assess the impact of a 4-day rural placement in Western Australia on the interest of fourth year medical students in a career in rural general practice. Students undertaking their Alternative Curricular Options in Rural Networks attachment (ACORNS) completed pre- and post-questionnaires examining their expectations, experiences of, and attitudes to rural general practice. Of the 103 students who participated, 81% expressed an interest in a rural career after the placement, whereas prior to this experience only 48% had been interested. The students also recorded a wide range of learning experiences, both clinical and procedural, and expressed positive attitudes to the variety of experiences and the role of the rural GP. The study concluded that early exposure to rural general practice enhances students' interest in a potential rural practice career and provides them with a broad range of experiences. The role of rural practitioners as role models for students needs to be acknowledged and reinforced. PMID- 11040576 TI - Rural and remote oral health, problems and models for improvement: a Western Australian perspective. AB - Oral healthcare in rural communities shares many of the dilemmas faced by medicine in providing services to large geographical areas with dispersed populations. This study examined the population data and service provision data relevant to the geographical distribution of oral health care in Western Australia (WA). Of the 1.7 million people resident in WA, 72% were resident in the five major urban centres with only 13% in rural and remote regions. Of the 320 postcode regions, 186 had a population of less than 2500, 31 had a population from 2500 to 5000, 42 from 5000 to 10,000, 37 from 10,000 to 20,000, and 24 had a population greater that 20,000. Almost 80% of postcode regions with a population less than 2500 are in non-urban regions. Of the total of 690 dentists who were analysed in this study, it was found that the vast majority (greater than 85%) worked in practices in postcode regions within metropolitan Perth or the major urban centres. A total of 43 postcode regions did not have a dental practice within their bounds. In order to address this disparity in service availability, strategies including the development of training for medical practitioners and auxiliaries, the use of modern technology, school-based programs and the development of interdisciplinary links should be implemented. These strategies would also facilitate the development of closer links between medical and dental practitioners and the development of skills within the medical fraternity that would facilitate improved oral health in rural and remote communities. PMID- 11040577 TI - Model of data structure and flow in general practice: a guide to evaluation of practice management software. AB - Despite its benefits, the computerisation of Australian general medical practices has been delayed, partly due to user concerns about the format of data dictated by available software and the portability of such data. This article proposes certain essential criteria for practice management software, which may provide a framework for its evaluation by users. A set of simple guidelines for database maintenance is also proposed. The author believes that users should become familiar with basic software architecture and drive the market to develop packages that conform to these minimal criteria. PMID- 11040578 TI - Palliative care on Kangaroo Island. AB - Kangaroo Island is the third largest island of Australia and the largest island off the coast of South Australia. A cross-sectional survey was conducted of professional carers and closest caring person of 15 patients who had died of malignant disease between 1994 and 1996. There was a high level of satisfaction with services provided on Kangaroo Island. There was regret about over-optimistic prognoses being given by specialist medical practitioners and considerable dissatisfaction with the service provided by tertiary hospitals. A divergence in assessment of symptoms was revealed between carers and professionals. There is need for improved training opportunities for community nurses, improved financial support for families dealing with malignancy and a need for more formalized bereavement services. PMID- 11040579 TI - Empowerment through information: supporting rural families of oncology patients in palliative care. AB - A research project examining the support needs of families caring for a relative in palliative care was conducted in New South Wales in 1997. Data were collected from 19 families and 10 specialist palliative care nurses from eight centres throughout New South Wales using audio-taped interviews. The findings show that information was one of the most important support needs for families to enable them to effectively care for their sick relative. The information provided needed to meet the individual and varying needs of families at different stages of their palliative care journey. Often the families described not knowing what their information needs were until a crisis occurred. Lack of information on a range of areas, from practical resources to providing physical care and managing medications, was problematic. Health professionals need to take responsibility in determining if families are aware of the information available, identify the most appropriate medium for this material and provide it at the pace and time wanted by the individual family. PMID- 11040580 TI - Issues influencing the provision of palliative care services to remote aboriginal communities in the Northern Territory. AB - Palliative care is a service available to many Australians subject to location and varying criteria. This paper seeks to identify some of the issues that hamper the provision of this sophisticated service to Aborigines living in remote parts of the Northern Territory. The paucity of literary sources of information demonstrates the need for research to be carried out in this field. PMID- 11040581 TI - Health and the 1999 Regional Australia Summit. AB - The Regional Australia Summit brought together 282 invited delegates from all parts of Australia. The aim of the Summit was to develop partnerships between the government, business and community sectors to deliver a better future for regional, rural and remote areas facing significant change. Health was one of 12 themes discussed at the Summit. Five key health priorities were identified; the need to change the dominant metropolitan mind-set, improve access to health-care services, improve service provision and workforce training, ensure equitable resource allocation, and adopt a population health approach. The ultimate success of the Regional Australia Summit will be gauged over time by the extent to which the health, wellbeing and prosperity of rural, remote and regional Australians has been improved, and existing problems and issues addressed. Nonetheless, the Summit is a significant event because it addresses issues at the highest level of government, emphasises coordination and the adoption of an intersectoral approach, and recognises the need to empower local communities and build partnerships between the government, corporate and community sectors. PMID- 11040582 TI - One voice regarding the legalisation of abortion. Nurses who experience discomfort. AB - The legalisation of abortion has been implemented since February, 1997. No research has been conducted on the effects of abortion on nurses. The objectives of this project were: 1. to explore and describe nurses' experience of abortion, and 2. to describe guidelines to support nurses. A qualitative, explorative, descriptive and contextual research design was used. The research was conducted in two phases, i.e., the exploration of nurses' experience, and description of guidelines. Trustworthiness measures as well as ethical measures were applied. Five themes were identified from the result of the interviews with nurses: freedom of choice of nurses to provide support; negative perceptions regarding the women who requested the abortion and staff providing the abortion; need for information; turmoil regarding life versus death, and recommendations from nurses regarding the management of abortion. Guidelines to support nurses were deducted from the identified themes. PMID- 11040583 TI - A comparison of a manual and a computer system in a primary health care clinic. AB - Previous research has shown that nurses providing primary health care in local clinics are burdened with extensive administrative duties. These administrative tasks are so time consuming that less time can be devoted to direct patient care, their primary function. Certain assertions have been made with regard to computerising the total environment of patient records, capturing statistical indicators, stock control, etc. It is said that computerising the clinic at the point of service will naturally lead to less time devoted to administrative tasks, thus increasing the time available for direct patient care. This study was done to determine whether this assertion is true. It was conducted by means of a combined quantitative and qualitative research design. The manual system and a computer system were quantitatively compared by means of various time measurements. The perceptions of patients and staff regarding computers were explored qualitatively by means of questionnaires. It was found that computerising the nurse's consulting room does not enhance the ratio between the time devoted to administrative tasks and the time devoted to patient care. In fact, the consultation time was longer with the computerised system than with the manual system. Some limiting factors in the computerisation process were discovered, the most important of which were the lack of computer literacy and typing skills of the nursing staff. PMID- 11040584 TI - [Nursing knowledge and attitude of day care personnel towards health promotion of babies]. AB - Currently it is more acceptable that women work. Besides women who are obliged to work to contribute to the income of their families, a large number of women work due to a need for social stimulation. While their mothers are at work during the day, the babies are left in substitute care out of necessity. Health promotion of babies is necessary and is the duty of the substitute caregiver. These women, however, do not always have the knowledge of, or positive attitude towards, health promotion. The purpose of this study, was to determine the knowledge and skills that the day mother should have in order to promote the health of babies (age 0-12 month). The qualititative research approach was used, within which a descriptive study had been done. A comprehensive literature study had initially been done regarding the health promotion of babies in substitute care. A programme had been compiled regarding the knowledge and skills that a day mother must possess in order to promote the health of babies. This program was then submitted to a group of selected experts for evaluation according to the Delphi technique. According to the data, it is quite clear that the day mother should be knowledgeable and skilled in order to satisfy the baby's physiological, security and affiliation needs as well as the need for appreciation and self actualization. PMID- 11040585 TI - Leadership development in a nursing service. A South African ethnographic perspective. AB - Leadership development at operational level in a nursing service is important to cope with all the transformation required. The purpose with this study is to explore and describe the views of black professional nurses on leadership behaviour, within the context of an operating theatre department, and to determine the influence of their occupational history on these views. An exploratory and descriptive research design, with an ethnographic perspective, was employed by means of semi-structured interviews (views on leadership) and questionnaires (occupational history). The results compare positively with the theoretical description of leadership behaviour. Guidelines for leadership development of these nurses were formulated, and should be followed by the necessary capacity building regarding leadership practice by professional nurses. PMID- 11040586 TI - Prevalence of depression in a university population. AB - OBJECTIVES: To determine the prevalence of depression in a university student population attending the Unitra Health Service and further compare the prevalence rate according to gender, age group and presenting complaint. METHOD: The sampling was carried out at the University of Transkei Health Service (UHS), Umtata. Two hundred and fifty students were randomly selected over a two month period during their first visit to the health centre for various ailments including family planning. Each student was given the 21-item Beck's Depression Inventory (BDI) questionnaire to fill at their own leisure time. RESULTS: The results of the study demonstrated a high prevalence of depression among the selected student population. BDI scores showed a total percentage of mild to severe depression to be 53%. BDI scores also showed that depression affects all age groups in this population with females being more affected than males at the ratio 3:1. All moderately and severely depressed students (14%) presented with somatic symptoms of headache and stomachache. None of these depressed students were diagnosed as being depressed by the UHS staff. CONCLUSION: Depression occurs more in females than males; it affects all age groups; is difficult to diagnose, even when it is severe, because of its somatic presentation. RECOMMENDATION: UHS nursing staff as well as students should be educated about depression to improve recognition and diagnosis. This recommendation applies to all nurses working at primary health care centres. PMID- 11040587 TI - Urban community health workers: selection, training, practice and outcomes. AB - The role, desirability and success of community health workers is debated. Conflicting reports have highlighted important concerns and provided guidelines. Particular issues identified are the necessity for both community and health professional input to determine needs and to ensure an acceptable selection process, training, support and accountability. Such steps were followed in the Greater Soweto Maternal Child Project. These are described together with the results achieved. Eight trained Soweto community health workers centered at Chiawelo Clinic and providing home based and neighbourhood health care undertake supervised Tuberculosis treatment, tracing of immunisation defaulters, and health education based on GOBI FFF (Grant JP, UNICEF:1985;94) and "Facts for Life" (UNICEF 1989-1993). They form a link between the community and government health care services and also other available resources. Over a period of 26 months, working from their own homes, they provided 14,254 health related services and in addition undertook 14,501 neighbourhood home visits. They were responsible for 8,710 referrals to the clinic or other relevant agencies for assistance. Incremental training has included HIV/AIDS counselling, advice on family planning with regular report back sessions and discussions. Participatory management involves all major role players. The community health workers have the approval and support of the Local Soweto Health Authority, the Civic Association and the communities they serve. On completion of the project, all were redeployed into local health service posts where it is intended that they form the nucleus of an expanding service. Delegation of selected tasks allows for cost effective functioning of more highly trained staff, an improved service and better use of available resources. PMID- 11040588 TI - [Support for the psychiatric nurse specialist and the psychiatric community nurse in their interactions with the psychiatric patient. Part II]. AB - In this article the research is described that had as goal to generate a supportive approach for the psychiatric nurse specialist to the psychiatric community nurse in interaction with the psychiatric patient, to promote, maintain and restore their mental health as an integral part of health. Guidelines for operationalisation of this supportive approach by the psychiatric nurse specialist, are also described. The research design utilised a qualitative, descriptive and contextual design. The exploratory field work was done in phase one of this research (as described in part I of these articles) and consisted of phenomenological interviews and focus groups. Trustworthiness was ensured by utilising Guba's model for trustworthiness. PMID- 11040589 TI - [Implementation and evaluation of supportive approach of the psychiatric nurse specialist and psychiatric community health nurse. Part III]. AB - The psychiatric community nurse takes on the role of an expert in her interaction with the psychiatric patient and others because she feels unprepared to manage therapeutic interaction. She measures her own abilities against the patient's progress and this makes it important for therapeutic interaction to succeed. This is accomplished when she exercises control over the interaction, manipulates the interaction by describing or reformulating the negative experiences or meanings verbalised by patients. She also distances herself from the psychiatric patient by stigmatising the psychiatric patient or if she feels that she cannot solve the problem, she refers the psychiatric patient to an external resource. The above narration indicates that this action by the psychiatric community nurse is not facilitative for patient interaction. In order to facilitate patient interaction, a supportive approach was generated and described by Van Wyk (1995: 99-116) whereby the advanced psychiatric nurse (a person with a masters degree or an advanced diploma in psychiatric nursing), creates a context where the psychiatric community nurse can sacrifice her objective observing position for an observed position where she forms part of the story of interaction between herself and the psychiatric patient. The objective of this study was to describe the implementation and evaluation of Van Wyk's (1995: 99-116) supportive approach to the psychiatric community nurse in interaction with the psychiatric patient by utilizing multiple descriptive case studies. PMID- 11040590 TI - Traditional birth attendants in South Africa: professional midwives' beliefs and myths. AB - It is necessary to establish in a scientific way what the knowledge of midwives regarding TBAs are, because misconceptions can very often lead to a negative attitude. The objective of the research was to determine what the practices of traditional birth attendants are regarding antenatal, intrapartum, and postnatal care according to professional midwives. An exploratory research design was used, where qualitative research methods were used to explore the above-mentioned aspects about the traditional birth attendant in certain communities in South Africa. Open-ended questionnaires were used to determine how midwives see the role of the TBA. Content analysis has been used to analyse the responses of the professional midwives. The role of the TBA according to the midwives can be summarised as follows, she is a middle aged or elderly lady with no formal training, who acquired her skills through experience. She attends to women during pregnancy, labour and the postnatal period in different ways that have been used by the TBA are mainly herbs that facilitate a quick delivery, or others that cause bleeding of the uterus postnatally. Other TBAs use no medicine. All the functions identified by the midwives were compared with literature about the role of TBAs in Africa. The majority of functions identified by midwives were found in the literature. The majority of professional midwives thought it a good idea to have TBAs who have undergone some form of training in order to improve their standard of practice and to teach them the early symptoms of complications. Implications for future practice will therefore be, amongst others, to start with training programmes for TBAs. An assessment in each community of the best way to do so, should first be done. PMID- 11040591 TI - The competencies of newly qualified nurses as viewed by senior professional nurses. AB - The information gathered from an in-depth literature study on the competencies of newly qualified nurses, was used to compile questionnaires for investigating newly qualified nurses' (NQNs') competencies as perceived by senior professional nurses (SPNs) in the former areas of Venda, Gazankulu and Lebowa (forming part of the Northern Province of the RSA since April 1994). The total number of 396 SPNs, employed by the health authorities in these three areas during 1994, comprised the population for this study. Questionnaires were distributed to the total 396 SPNs and responses were received from 259 SPNs (implying a response rate of 65.4%). Application of stages of the nursing process (problem solving and clinical judgment), research, management and administration of a clinical unit, nursing ethics and critical care were perceived by the SPNs to be the central focus of NQNs' incompetency in all four clinical nursing units, namely community, psychiatric, midwifery, and general units. The findings provide nurse educators and curriculum developers with realistic input about the SPNs' expectations from NQNs in the real working situation. This information could assist in the delineation and refinement of the professional competencies expected of nurses trained in the comprehensive (R425) course. PMID- 11040592 TI - The living conditions of elderly black people in a typical rural area in the Free State. AB - The proposed policy in respect of the care of older persons places much emphasis on the involvement and responsibility of the community. Majwemasweu is a typical rural black community in the Free State where an active Committee for the care of older persons is already in place and has already largely taken the responsibility for caring for its older persons on itself. This research scrutinizes the living conditions of this community's older persons and, wherever possible, compares it with those of black urban and rural research groups in the Multidimensional Survey of Elderly South Africans, 1990-1991. Bearing in mind the time lapse between die two surveys, the situation of the Majwemasweu group is in many aspects much better than those of the two groups with which it is compared. Their socio-economic position is, in many respects, more favourable as a result of political change, when compared with the two other groups, while the unremitting zeal of the local Committee for the care of older persons has undoubtedly contributed to their more favourable position. It would appear that the community is ready for the implementation of the new policy for the care of older persons. Internal political disputes which have already been responsible for the retraction of sponsorship for upgrading the community health clinic, will, however, have to be settled immediately. Sustained intervention through dialogue on the side of the researchers continues to attempt to stimulate the development potential of the community with a view to successfully implementing the new policy. PMID- 11040593 TI - Satisfaction with family planning services. Interpersonal and organisational dimensions. AB - In South Africa, client satisfaction with the quality of health care has received minimal attention; probably due to the lack of locally developed and tested measures. Therefore, we developed and tested a 20-item attitude scale to determine satisfaction with Family Planning (FP) services. The objectives of this study were to: ascertain reliability of the scale and confirm, through factor analysis, that satisfaction with the FP service was based on interpersonal and organisational dimensions. The sample comprised 199 black adult interviewees (158 women and 41 men), who had previously used or were currently using contraception, from an informal settlement in Gauteng, South Africa. Three items were removed from the scale due to unacceptable communality estimates. The reliability coefficient of 0.76 for the 17-item scale was satisfactory. The principal components analysis, with orthogonal and oblique rotations, extracted two factors; accounting for 51.8% of the variance. The highest loadings on Factor I involved an interpersonal dimension (friendly, encouraging, competent, informative and communicative). Factor II tended to focus on the organisational elements of the system, such as different methods, choice of methods, service availability and length of waiting time. It was concluded that this scale was a reliable, easily administered and scored measure of satisfaction, with underlying interpersonal and organisational dimensions. PMID- 11040594 TI - The integration of the rehabilitation of psychiatric patients into the primary health care system. AB - This study done in rural and semi-urban clinics examined the ability of primary health care nurses in providing rehabilitation of psychiatric patients in the Primary Health Care service. The objectives of the study were to train and evaluate registered nurses' ability to implement rehabilitation to psychiatric patients in the community. Registered nurses were trained over a period of 10 days. Each client who visited the clinic had a rehabilitation plan drawn with the client and family. Families participated in the training of clients while nurses were trained to identify target symptoms, draw a plan to be followed by the client and his family, set rehabilitation goals and the steps to achieve the goals. The project was implemented over a period of 12 months. Records were then reviewed 1 year after implementation and at 18 months to determine the performance of nurses. Evaluation was done per clinic. Each clinic was evaluated and differences were found and where performance was poor, re-education was done. Each clinic was seen as a case. A record review was done to determine the level of rehabilitation based on the following: identification of target symptoms plan for the patient plan for the family setting of rehabilitation goals steps to achieve goals level of vocational rehabilitation. PMID- 11040595 TI - [Grievances in South African hospitals: from a nursing service management perspective]. AB - The nursing service manager is accountable for adequate and efficient personnel management in the nursing service and the management of grievances is an important aspect of the personnel management function. The question arises, however, how and when grievances in nursing services arose and developed? The purpose of this article is to give a historical description of the development and management of grievances in nursing services for the time frame 1652-1990. An historical analysis was undertaken by means of news paper analysis, as well as other written resources. The results show that the development of grievances are related to the development of hospitals in South Africa and that grievances were poorly managed. The following conclusions are made: grievances in nursing are related to the establishment of hospitals; the first official grievance was lodged in 1824 by a surgeon; grievance are mainly related to the working conditions, remuneration and management; complaints with salaries and food were lodged by nurses as early as 1869; it appears as if nursing service managers are not adequately skilled in the management of grievances experienced by nursing staff--the same mistakes are made leading to strike action by nurses/midwives; unhappiness with the inappropriate manner in which grievances are managed lead to industrial action by nursing staff since 1889. Continuous empowerment of nursing services managers in the management of grievances is important and therefore the development of a model for grievance management in nursing services is also recommended. PMID- 11040596 TI - A feasibility study of part time day and evening classes in the Nursing Science Department of the University of Zululand. AB - The study determines the practising professional nurses including those in the nursing colleges who would enroll if part time studies were introduced at the University of Zululand in 1991. Questionnaires were sent to various institutions in KwaZulu-Natal. Four hundred and fourty eight (448) nurses responded positively to the questionnaire which was an indication that if such studies were introduced many nurses would enroll. The study reveals the enthusiasm and action taken by a nurse who is working and attending classes part time. PMID- 11040597 TI - Legal limitations for nurse prescribers in primary health care. AB - The nurse plays an important role in the delivery of primary health care services in South Africa. The primary purpose is to provide the public with access to safe competent basic health care and to achieve this, the nurse should be empowered to practice within legal and ethical boundaries. This paper explores and describes the limitations imposed by legislation on the nurse's ability to prescribe treatment in the primary health care field. The focus is mainly on the Nursing Act, the Pharmacy Act and the Medicines and Related Substances Control Act which highlights a number of limitations. It is concluded that empowerment of the nurse should not only include addressing the legal boundaries for practice, but also education and training opportunities to equip them with the expert knowledge and skills that they need to render a quality health care service. PMID- 11040598 TI - [The phenomenon of families who are involved in decision making about life support withdrawal in family members]. AB - The overall objective of this study was to explore and describe the phenomenon of families who are involved in deciding about withdrawal of life-support treatment of a family member. A phenomenon analysis was undertaken in two phases. During the first phase, secondary analysis of primary data was done on the family used in Burger's study (1996: 1-175) and was followed up by phenomenological interviews with families used as member checking from the same circumstances and according to the same criteria that Burger (1996: 1-185) used in her study. Data were analysed in collaboration with an independent coder. The family used as member checking in this study is also used as data control. A literature control was conducted as part of data control. The themes that were identified included were: physical and bodily experiences of families; defence mechanisms used by families to cope with grief; emotional experiences of families; need of knowledge/perceptions/outlook on life/internal conflict/feelings of guilt/ability to make decisions/respect of patient wishes/the effect of time and prior experiences; support needed by an given to families; spiritual and supernatural experiences/hope/acceptance/ability to 'let go' of the patient. In phase two, guidelines were described for psychiatric nurse specialists to mobilise resources for families to promote, maintain and restore their mental health as an integral part of health. PMID- 11040599 TI - Contribution of unit managers to the training of student nurses in the Cape Peninsula. AB - The article is based on research conducted over the period 1993 to 1996 in the Cape Peninsula. The purpose of the study was to determine the contribution of unit managers towards the training of student nurses coming to their units for clinical practica. The sample consisted of student nurses training in the four nursing colleges in the Cape Peninsula, and the unit managers working in the health services accommodating students for clinical practica in the same area. The findings revealed that the majority of unit managers were teaching students whenever they had the opportunity. Generally unit managers were prepared for their teaching function, but many students were not satisfied with some clinical learning opportunities presented to them, for example drawing up patient care plans, discussing patients' treatment plans when handing over report, giving assistance regarding care decisions and lending support when students are confronted with patient care problems. There appears to be a need to educate unit managers regarding these and other aspects of the students' training programme. PMID- 11040600 TI - Nursing professionals' views on the workplace. Survey results. AB - This article reports on a survey done among registered, enrolled and auxiliary nurses registered with the South African Nursing Council. The survey was carried out in the period from the end of December 1997 to the beginning of 1998. The purpose of the survey was to obtain the views of female nurses on various aspects of the workplace. The important findings were the fact that nurses liked working as part of a team and that this contributed the most to their job satisfaction. The item that contributed least to job satisfaction was pay. The most important problems were that they felt that they were not paid enough and that they need better benefits. The majority of nurses were however positive about their jobs and the items the highest on the list of career expectations were job satisfaction, followed by a need for recognition. PMID- 11040601 TI - [Financial advocacy: a true reality?]. AB - It seems that in spite of soaring medical costs and a poor economic climate, financial advocacy for patients is not general practice. A question that arises is what part nurses play and what their perceptions on financial advocacy for patients are. A non-experimental descriptive study was done to determine what nurses' perceptions on financial advocacy are and how knowledgeable they are about the costs of disposable dispensary stock. Thirty patient accounts were analysed and questionnaires were completed by a convenience sample of thirty two registered nurses who had permanent appointments in high care or intensive care units. RESULTS: On average nurses control the utilization of 15.8% of each of the prescription accounts. Only 12.5% of respondents had formal education on their responsibility in keeping costs low. The majority (60%) of the 50% of respondents who had previously received information had performed financial advocacy although 94% agreed that they have the responsibility to do so. A mere 9% was achieved on the knowledge of the costs involved in using consumable products from the dispensary. PMID- 11040602 TI - The cost-effectiveness of managed care regarding chronic medicine prescriptions in a selected medical scheme. AB - The purpose of the study was to examine the cost-effectiveness of managed care interventions with respect to prescriptions for chronic illness sufferers enrolled with a specific medical scheme. The illnesses included, were epilepsy, hypertension, diabetes and asthma. The managed care interventions applied were a primary discount; the use of preferred provider pharmacies, and drug utilization review. It was concluded that the managed care interventions resulted in some real cost savings. PMID- 11040603 TI - Establishing a South African Honour Society of Nursing under auspices of Sigma Theta Tau International (Inc.). PMID- 11040604 TI - A management model for professional wholeness. AB - The purpose of this article is to describe a management model for professional wholeness. A theory generating research design was followed. Based on this model the consequences of management could be a state, which varies dynamically between minimal and maximal professional wholeness. A list of descriptive words was developed to generate qualitative indicators of professional wholeness. PMID- 11040605 TI - [Endometriosis in tampon users]. AB - Endometriosis has been recognized as one of the possible causes for infertility. If endometriosis occurs more often in a women who uses tampons comparing to those who do not, it is an important point of discussion to the modern women and nurses. Recent research (van Rijswijck & Botha, 1997) indicated that a correlation between the use of tampons during menstruation and endometriosis exists. It seems that an educational program, by primary health care workers, indicating the potential dangerous effects of tampon use will lower the incidence of endometriosis, thus the problems with infertility could lesson and optimal health care to women could be improved. PMID- 11040606 TI - A procedure for evaluating primary health care software. AB - Managers in health care often find themselves in the difficult position of having to make decisions regarding the purchasing of software and hardware which they are not qualified to make. The aim of this paper is to support health managers in their decision making by means of a procedure and an instrument that can be used to evaluate primary health care software. A seven step approach to the evaluation process is proposed and each step is discussed in detail. The paper concludes with a proposed software evaluation instrument that is suitable for application in the health care environment. PMID- 11040607 TI - Nursing research: can a feminist perspective make any contribution? AB - As more than 90% of the RSA's nurses are women and as at least 50% of the health care clients are also women, nursing research can definitely benefit by incorporating feminist research approaches. Specific feminist research issues which could be relevant to nursing research include: inherent themes in feminist research feminist research methodology gender stereotypes and nursing research gender-based stereotypes of researchers potential benefits of incorporating feminist research approaches in nursing research. Most formal models of nursing, and thus also most nursing research based on these models, ignore gender issues. Thus they ignore part of the social reality of nursing and might provide distorted images of nursing. A feminist approach to nursing research could enhance the reality-based gender issues relevant to nursing specifically, and health care generally, and contribute towards rendering effective health care within a multidisciplinary health care context. PMID- 11040608 TI - [Nurses' perceptions of inservice training in a private hospital]. AB - A private hospital aims at quality health care delivery with a profit making motive. Inservice education to nursing staff therefore plays an important role- it should add value to the hospital and should also be cost-effective. This research hospital has a formal inservice education programme for the professional nurses, but the question arises: what are their perceptions of the inservice education in this hospital? The aim with this research is to explore and describe the perceptions of the professional nurses on inservice education and to formulate guidelines for improvement. A qualitative research strategy was utilised, with individual focus interviews as method of data collection, followed by a content analysis of the transcribed interviews. Two main categories were identified, being the positive or facilitative perceptions and the negative perceptions/obstacles. Concluding statements were formulated to serve as basis for the description of guidelines on inservice education within this hospital. Guidelines for quality inservice education were formulated, relating to the following: scientific principles to make provision for appropriate assessment, planning, implementation and evaluation of the inservice education programme; accessibility of the inservice education sessions; establishment of a positive learning climate by the learning accompanist; credibility of the inservice education programme in the private hospital. It is recommended that these guidelines be implemented and that selected statements be converted to hypotheses and exposed to testing. PMID- 11040609 TI - Stimulating community participation in a group of farm workers using action research. AB - This study assessed whether community participation in health related activity was a reality or just popular development rhetoric. Using action research methodology, focus group discussions and informal contacts were made with farm workers consisting of twelve families in Umkomaas, south of Durban in the province of Kwa Zulu Natal. The aim was to establish whether this community could be actively involved in all aspects of community participation. The level of participation was described using Rifkin's model (Rifkin et al, 1988). Results of this study revealed that the community was able to identify their own health problems, prioritize them and plan appropriate strategies to meet the needs identified. PMID- 11040610 TI - The use of reproductive health information material in the rural clinics of Umtata district. AB - Though it appears that there has been a long history of development, distribution and dissemination of reproductive health information materials, impediments still exist in the health information, communication and education systems in South Africa. The aim of the study was to contribute towards improvement of the development, distribution and utility of reproductive health information material both in the Eastern Cape province and nationally. In-depth understanding of complexities surrounding development, distribution and utility of educational material is key to the provision of information to communities. The objectives were to describe the availability and examine the content, target groups, language and utility of reproductive health information materials. Qualitative study using non probability sampling was done. Ten rural clinics were conveniently selected. In-depth interviews, focus groups and a checklist were used to collect data. Reproductive health information materials found were mostly posters, there were no pamphlets, material on some reproductive health aspects was not available, inconsistently distributed and not updated. It was concluded that posters are available in the rural clinics of Umtata district judging from the results there is a need to improve the quality of reproductive health information material. PMID- 11040611 TI - Teenage mothers' knowledge of sex education in a general hospital of the Umtata district. AB - There has been growing concern about the increase in teenage pregnancies in relation to the teenagers' knowledge of human sexuality and the impact sex education has on these teenagers in both the urban and rural areas. The aim of the study was to assess the knowledge of sex education and the health beliefs of teenagers with regard to teenage pregnancy. A descriptive study was conducted in the Umtata district of the Eastern Cape. The sample involved 42 teenage mothers drawn from local rural and urban areas attending a Well Baby Clinic at Umtata General Hospital. A questionnaire was used as the method of data collection. Data analysis was done by a software package called SAS. The study revealed that teenagers receive almost no sex education from health personnel and only a little from their parents. The study also revealed that most of these teenagers live with their mothers only instead of both parents. It also became clear that unsafe or unprotected sexual behaviour was practised by these teenagers although teenagers supported the idea of their partners using condoms. The most common problem resulting from teenage pregnancy, as indicated from the study results, was the financial burden on parents and lost educational opportunities by the girls. In the recommendations the parents' involvement in sex education and the improvement of recreational facilities for both urban and rural areas are highlighted. In conclusion, the study has shown the need for more efforts to solve the problem of inadequate sex education and to change the health beliefs of teenagers. PMID- 11040612 TI - Nursing ethics in a developing country. AB - Nursing is a true profession, distinguished by its philosophy of care, its full time commitment to human wellbeing, its particular blend of knowledge and skills and its valuable service to the community (Curtin & Flaherty, 1982:92). Ethics is vital to nursing. Being a professional implies ethical behaviour and knowledge of what it means to be ethical (Pera & Van Tonder, 1996:v). Ethics is the foundation of committed service to humankind. When nurses practice is an ethical manner they should adhere to ethical principles like autonomy, beneficence, justice, veracity, fidelity, confidentiality and privacy. From this conceptual framework two questions can be asked, namely: Does the behaviour of nurses in health services in South Africa comply with the principles of ethics? How can ethical behaviour be facilitated in nurses in South Africa? The first question was answered by doing a critical analysis of thirty-two case studies of recent ethical phenomena in health services. The ethical principles will be used as criteria for this analysis. Some of the ethical case studies will be presented in this paper to indicate the problems in relation to autonomy, beneficence, justice, veracity and fidelity. It will be demonstrated that from deontological ethical theories nurses are not doing their duty as advocates for the vulnerable patient and from utilitarianism the poor and uneducated patients are being exploited. To empower patients in developing countries it is of vital importance for nurses to behave in an ethical manner. From a literature study a program for rational interaction for moral sensitivity (Rossouw, 1995) and virtue-based ethics in Nursing Education is identified to facilitate moral behaviour amongst nurses in developing countries. PMID- 11040613 TI - Clinical skills of nurses in mobile health services. AB - The purpose of this study is to explore and describe the acceptability of the clinical skills of community nurses in mobile health services. An explorative, descriptive design was employed. After a literature study, interviews were conducted with patients, and analysed, the results were verified by means of observation of the mobile services. The clinical skills were described as favourable and not favourable by patients some of which were confirmed during the observation phase. Guidelines for a more user friendly service were written. PMID- 11040614 TI - [Evaluation of arguments in research reports]. AB - Some authors on research methodology are of opinion that research reports are based on the logic of reasoning and that such reports communicate with the reader by presenting logical, coherent arguments (Bohme, 1975:206; Mouton, 1996:69). This view implies that researchers draw specific conclusions and that such conclusions are justified by way of reasoning (Doppelt, 1998:105; Giere, 1984:26; Harre, 1965:11; Leherer & Wagner, 1983 & Pitt, 1988:7). The structure of a research report thus consists mainly of conclusions and reasons for such conclusions (Booth, Colomb & Williams, 1995:97). From this it appears that justification by means of reasoning is a standard procedure in research and research reports. Despite the fact that the logic of research is based on reasoning, that the justification of research findings by way of reasoning appears to be standard procedure and that the structure of a research report comprises arguments, the evaluation or assessment of research, as described in most textbooks on research methodology (Burns & Grove, 1993:647; Creswell, 1994:193; LoBiondo-Wood & Haber, 1994:441/481) does not focus on the arguments of research. The evaluation criteria for research reports which are set in these textbooks are related to the way in which the research process is carried out and focus on the measures for internal, external, theoretical, measurement and inferential validity. This means that criteria for the evaluation of research are comprehensive and they should be very specific in respect of each type of research (for example quantitative or qualitative). When the evaluation of research reports is focused on arguments and logic, there could probably be one set of universal standards against which all types of human science research reports can be assessed. Such a universal set of standards could possibly simplify the evaluation of research reports in the human sciences since they can be used to assess all the critical aspects of research reports. As arguments from the basic structure of research reports and are probably also important in the evaluation of research reports in the human sciences, the following questions which I want to answer, are relevant to this paper namely: What are the standards which the reasoning in research reports in the human sciences should meet? How can research reports in the human sciences be assessed or evaluated according to these standards? In answering the first question, the logical demands that are made on reasoning in research are investigated. From these demands the acceptability of the statements, relevance and support of the premises to the conclusion are set as standards for reasoning in research. In answering the second question, a research article is used to demonstrate how the macro- and micro-arguments of research reports can be assessed or evaluated according to these standards. With evaluation it is indicated that the aspects of internal, external, theoretical, measurement and inferential validity can be evaluated according to these standards. PMID- 11040615 TI - [Evaluation of the quality of nursing care in a number of homes for the aged in Orange Free State]. AB - A study was undertaken to evaluate the quality of nursing care in a number of homes for the aged in the Orange Free State. Ten homes were visited and 45 frail aged patients observed. Data was collected by means of a standardised instrument. Essential physical needs such as hygiene and nutrition were found to receive the necessary attention. However, aspects such as stimulation, socialisation, reality orientation, habit training programmes and exercise did not receive enough attention. In the light of these findings it was concluded that a custodial care approach was followed in these homes. PMID- 11040616 TI - Perceptions of overweight African women about acceptable body size of women and children. AB - PURPOSE: Malnutrition, presenting as obesity in women and under-nutrition in children, is a prevalent problem in the squatter communities of Cape Town. Food habits are determined by a complex matrix of economic, social and cultural factors which need to be understood by health professionals prior to the implementation of strategies to improve the nutritional status of this community. This qualitative study is designed to explore the perceptions of overweight black women in Cape Town, with underweight infants, about the culturally acceptable body size for women and children. METHOD: Qualitative in-depth interviews were conducted with 10 overweight black women who were resident in the metropolitan area of Khayelitsha in Cape Town. A snowballing technique was utilised to select the key informants, all of whom were mothers of underweight infants. The interviews were conducted in Xhosa and recorded, with the permission of participants, onto tape. They were then transcribed and translated into English. The transcripts were coded and analysed by two researchers who worked independently to ensure content validity. RESULTS: The informants came from disadvantaged communities in which food was highly valued as a result of the fact that food security was not assured. The concept of an individual voluntarily regulating the intake of nutrients when food did become available, appeared unacceptable to the informants. It was not clear from the interviews how the participants perceived their normal or "desired" body weight. Increased body mass was regarded as a token of well-being in that marital harmony was perceived to be reflected in increased body weight. Overweight children were regarded as reflecting health as it was associated with sufficient food supply and intake. CONCLUSIONS: Although women expressed the desire to loose some excess weight for practical reasons, there was no negative social pressure to motivate this. The attitudes recorded from this qualitative research project suggest cultural perceptions of excess body weight that will complicate the design of effective health promotion strategies to normalise and maintain ideal body weight in this group of African women. PMID- 11040617 TI - The use of a rating instrument to teach and assess communication skills of health care workers in a clinic in the Western Cape. AB - Research in health communication shows communication to be an important aspect of successful health-care. Moreover, training courses which provide feedback have been shown to improve health professionals' ability to conduct successful interviews. This article describes a rating instrument which was developed in order to facilitate teaching and assessing the communication aspects of health care interviews. The instrument was found to be useful in a training programme offered to nursing staff of a TB Clinic in Mitchells Plain, Western Cape. The instrument appears as Table 1. In the Table categories of communication behaviours, each indicating an important aspect of the interaction, are given as the six headings. These are: establishing rapport and respect listening receptively confirming the patient sharing control informing effectively and checking perceptions. Within each category the more detailed specific behaviours are listed, allowing for close analysis of a care-giver's interviewing skill. The article briefly discusses the importance in effective communication between the care-giver and patient of each category of behaviours given in the instrument, supported by evidence from research. Lastly the article describes a "case study" on how the instrument has been successfully used in a training programme. PMID- 11040618 TI - The experience of education students in the TechnoLab. Do engineers and educationalists have to co-operate? AB - The purpose of this study was firstly to explore and describe post graduate students' experiences of Technology Education in the TechnoLab and secondly to deduct recommendations to assist and facilitate the learning process of students in the TechnoLab. The research design can be described as qualitative, exploratory, descriptive and contextual. Twenty-eight post graduate students in Technology Education participated in a workshop at the TechnoLab for one morning session. The aim of this workshop was to expose them to the so-called technological process, which they had already studied theoretically. After participating in this workshop students were requested to write down their experience of Technology Education in the TechnoLab. They were then divided into three focus groups for interviews conducted by three moderators. The same question was posed to each group: "How did you experience Technology Education in the TechnoLab?" These interviews were audiotaped and later transcribed. Tesch's method of analysis was applied to the data by three researchers independently followed by a consensus discussion. Three major themes were identified from the data, namely the TechnoLab and Technology Education were conducive to learning; fear and anxiety were experienced in the unknown situation, that is the TechnoLab and Technology Education; and empowerment through exposure to the TechnoLab and Technology Education. Further categories were identified within each of these three themes. Recommendations are made to assist and facilitate the learning process of these students in the TechnoLab. PMID- 11040619 TI - Perceptions of black women of obesity as a health risk. AB - This article focuses on the exploration and description of perceptions of patients and community nurses of the phenomenon of obesity as well as any potential problems that could be foreseen if health promotion programs were planned for obese people. Whilst the researcher sought to explore patients' perceptions of obesity as a threat to health it was also necessary to obtain similar information from the community nurses who were giving health services to these patients'. The level of knowledge of obesity as a health problem as well as the nurses personal attitude towards the phenomenon of obesity is crucial in the manner and quality of her interventions. A qualitative, exploratory, descriptive and contextual research design was utilised. Semi-structured interviews were conducted amongst patients and community nurses to collect data regarding perceptions of obesity. Data obtained from patients can be used meaningfully for the planning of health promotion programmes for the obese individuals and families. Some negative and ambivalent statements were also identified as problem areas that need to be worked on. Conclusions drawn from the findings on nurses responses highlighted a number of problem areas which were identified in the cognitive, psychomotor and affective domains of the community nurse as health professional. Certain shortcomings were also identified with the community health structures which may not be supportive to the community nurse, individuals and families with problems of obesity. PMID- 11040620 TI - Adolescents previously involved in Satanism experiencing mental health problems. AB - No research has previously been done regarding the phenomenon of adolescents who have previously been involved in Satanism and who experience obstacles in their strive for mental health. Adolescents previously involved in Satanism present behavioral problems like aggressive outbursts, depression, "psychosis" or suicide attempts, that could lead to suicide. In the phenomenon-analysis semi-structured, phenomenological interviews were performed with the respondents and their parents. The respondents were requested to write a naive sketch about their life. After completion of the data-control, guidelines for nursing staff were set. The guidelines are set for the management of adolescents who have previously been involved in Satanism and who experience obstacles in their strive for mental health. Interviews with experts in Satanism were conducted, literature in the form of books, magazines and newspaper-clippings were used to verify the research findings. The most important guidelines are that the caregivers have to be reborn Christians; they are not allowed to show any fear or sympathy; they must have sufficient knowledge about Satanism; the adolescents have to be unconditionally accepted; the caregivers have to work in a team and the adolescents have to be taught to deal with their emotions. PMID- 11040621 TI - Furthering caring through nursing education. AB - The nursing students' main quest is for self actualization by attributing meaning to life through caring. To assist student nurses in this quest, the nurse educator needs to plan educational interventions according to an anthropological model that posits care and caring as innate human attributes. Further, the structural essence of what professional nursing caring entails should also be posited as a point of departure for curriculum planning. The author proposes such models. The main implications include that the nursing curriculum must increasingly attend to the emotional needs of nursing students. Curricular content and teaching strategies toward this goal are suggested. PMID- 11040622 TI - Parasuicide among youth in a general hospital in South Africa. AB - Parasuicide cases among youth (15-24 years) referred to the clinical psychology section of a regional hospital from 1995 to 1998 were reviewed. In all 100 cases (37 males and 63 females) were identified being about 10% of the caseload. As part of the clinical psychological assessment sociodemographic, clinical characteristics, trigger factors, employed methods and suicide intentions were analysed. Most patients were students (79%) or unemployed (16%). The major method employed to attempt suicide was ingestion of harmful substances (like paraffin, pesticides or battery acid)(73%). Acute social conflicts (38%), socio-economic deprivation (17%), AIDS phobia (17%), academic failure (14%), teenage pregnancy (10%) and mental illness (5%) triggered suicide attempts. Fifty-eight percent of the attempts were categorised as demonstrative and 27% as genuine. The psychodynamics of parasuicides are discussed in case studies and with reference to other studies. PMID- 11040623 TI - A development and support programme for black first year nursing students. AB - The changed student culture in the School of Nursing from a traditional white, Afrikaans-speaking to a multicultural student community, led to a variety of problems. Many of these were experienced by students (mainly those in their first year) from historically disadvantaged educational backgrounds. The aim of this study was to establish a developmental and support programme for black first year nursing students. An action and developmental approach was followed and the research was carried out over a period of four years. The programme was developed and evaluated by means of several actions in Phase I (1992-1995) and re-planning took place throughout. The programme was implemented in Phase II and the level of student development was determined. The students experienced a number of problems and their performance as regards development was below average in terms of cultural participation and interpersonal relationships. In spite of the fact that the majority had commenced their studies with an average E matriculation symbol, 84.8% of them passed five or more semester courses at the end of 1996. PMID- 11040624 TI - The critical thinking ability of diplomates from different types of bridging programmes. PMID- 11040625 TI - Experiences of student nurses in a multicultural nurse-patient encounter. AB - South African nurses have accepted the challenge that was brought about by the Health Reform Policy of 1990 which decreed that health service centres be opened to people of all cultural and racial groups. However, studies on transcultural; nursing have revealed that, problems have occurred during a multicultural nurse patient encounter. Most of these studies have approached the problems from the patients point of view, this study was therefore an attempt to look into the problem from the nurses point of view. The researcher was interested in exploring the sources of such problems, their effects on the nurse patient relationship as well as to find out from the respondents, the possible solutions to such problems. Through focus group interviews (FGI), respondents who had experienced problems with culturally different patients were given an opportunity to reflect on those experiences, report on these and recommend possible solutions. The results revealed that differing perceptions about the encounter between the nurse and the patient, previous experience which led to the formation of stereotypes, was the major source of problems. The results further revealed that stereotype relevant information, if used during the nurse patient encounter, led to inadequate care delivery. The solution to the problem according to the results, would be a positive approach to the problem of cultural differences. PMID- 11040626 TI - [Concept analysis of reflective thinking]. AB - The nursing practice is described as a scientific practice, but also as a practice where caring is important. The purpose of nursing education is to provide competent nursing practitioners. This implies that future practitioners must have both critical analytical thinking abilities, as well as empathy and moral values. Reflective thinking could probably accommodate these thinking skills. It seems that the facilitation of reflective thinking skills is essential in nursing education. The research question that is relevant in this context is: "What is reflective thinking?" The purpose of this article is to report on the concept analysis of reflective thinking and in particular on the connotative meaning (critical attributes) thereof. The method used to perform the concept analysis is based on the original method of Wilson (1987) as described by Walker & Avant (1995). As part of the concept analysis the connotations (critical attributes) are identified, reduced and organized into three categories, namely pre-requisites, processes and outcomes. A model case is described which confirms the essential critical attributes of reflective thinking. Finally a theoretical definition of reflective thinking is derived and reads as follows: Reflective thinking is a cyclic, hierarchical and interactive construction process. It is initiated, extended and continued because of personal cognitive-affective interaction (individual dimension) as well as interaction with the social environment (social dimension). to realize reflective thinking, a level of internalization on the cognitive and affective domain is required. The result of reflective thinking is a integrated framework of knowledge (meaningful learning) and a internalized value system providing a new perspective on and better understanding of a problem. Reflective thinking further leads to more effective decision making- and problem solving skills. PMID- 11040627 TI - School and nursing service managers' ability to hold their own amidst daily demands. AB - Improving quality of life itself subsumes the experience, maintenance, and continuous enhancement of physical and psychological health. It is therefore important for the physical and psychological health of school and nursing service managers that they are able to hold their own successfully, both personally and professionally. Bearing this in mind, the question of the extent to which school and nursing service managers succeed in holding their own (coping) successfully remains. Does their profession (field of work) influence school and nursing service managers' personal beliefs, perceptions of their coping ability amidst daily demands and their use of different types of coping strategies. The purpose of this research project was to address these issues. An empirical investigation was undertaken with reference to a theoretical framework to assess school and nursing service managers' personal beliefs, perceptions of their coping ability amidst daily demands and the coping strategies they use. Consecutive factor analytical procedures, item analyses, Hotelling T2-tests and Student t-tests were conducted on the data. The results of the analyses indicated, inter alia, that nursing service managers and, especially school managers, do not quite perceive themselves capable enough to cope with interpersonal demands. School managers find it significantly more difficult than nursing service managers to cope with demands arising from professional and personal relationships. PMID- 11040628 TI - The role of the ward manager in creating a conducive clinical learning environment for nursing students. AB - Ward sisters/managers are without doubt the professional gate keepers of the ward environment yet there are activities in that environment for which they do not seem to take full responsibility, namely that of the clinical learning of nursing students. The aim of this study was to investigate the role of the ward manager in creating a conducive clinical learning environment for nursing students. An explorative descriptive research method was employed. Findings reveal that the ward managers are generally satisfied with the way in which they handle the important role they play in facilitating teaching and learning for nursing students. They feel strongly, however, that the nursing students themselves need to be active in the learning process. While acknowledging the efforts of the ward managers in creating and maintaining the learning environment, nursing students were dissatisfied about several aspects that appeared to be lacking in the clinical environment, such as good interpersonal relations, support, exposure to practice administrative skills (for example, problem-solving and decision-making) and lack of feedback about their performance. There appears to be a need to develop more effective support structures within the learning environment so that nursing students can obtain sufficient exposure to learning opportunities. PMID- 11040629 TI - The importance of biographic research: a South African black nurses' perspective. AB - This article is an attempt to highlight the importance of biographic research to South African nursing. The writer believes that a particular attention should be paid to the contributions of South African Black/African nurse practitioners. South Africa has produced remarkable African nurses: they range from nurse Professors and Head of the University Nursing Departments to clinical nursing specialists and nursing administrators. The writer--having used the biographical approach in his Doctoral thesis--will highlight some practical and professional issues around biographic research. For the purpose of this publication, however, discussion will be confined to defining biographic research, reviewing different types of biographies, and discussing the value of the biographical research. Furthermore, the writer will identify some biographic concepts, examine their relationships, draw inferences and (hopefully) emerge with an increased understanding of the impact of biography as scientific concept. PMID- 11040630 TI - [Standards for neonatal intensive care nursing: unit directed management standards]. AB - The neonate has the right to quality nursing care and the Neonatal Intensive nursing care practitioner is professionally-ethically and personally liable for quality nursing care. The process of quality improvement is a structured, planned and purposeful action were standards are set and the nursing care is evaluated after which remedial steps are taken to improve quality nursing care. In this study the focus is on the first step in the quality assurance cycle:the setting of standards. The purpose of the study is to describe and formulate standards for Neonatal Intensive nursing care which can be utilised as an accreditation instrument for institutional selfevaluation to improve quality nursing care. Standards for Neonatal Intensive nursing care were developed and validated by utilising a three-phase research method. In phase one subjects for standards were identified by a panel of experts. The identification was done by means of a critical debate, after which a preliminary conceptual framework was formulated. During the second phase a literature control was done to refine the conceptual framework. It consisted of a conceptual framework pertaining to unitmanagement (article 1) and an conceptual framework pertaining to high incidency, high risk interactions in the Neonatal Intensive Care Unit. The conceptual framework pertaining to the high incidency, high risk interactions in the Neonatal Intensive Care Unit consisted of: assessment of the neonate preparedness for neonatal resuscitation mechanical ventilation humidification during ventilation physiotherapy and suctioning of the ventilated neonate weaning during mechanical ventilation ecstubation During the third phase, the standards were validated by means of a consencus debate. An accreditation instrument was developed for institutional selfevaluation to improve quality nursing care in the Neonatal Intensive Care Unit. The standards that were formulated consisted of standards for unitmanagement (article 1) and standards directed at high incidency, high risk interactions in the Neonatal Intensive Care Unit which is published in the second article in this series. PMID- 11040631 TI - A model for culture-congruent psychiatric nursing. AB - The purpose of the research conducted was to explore and describe the influence of culture on approaches to mental illness and the patients' compliance with psychiatric treatment, and to generate a practice model for culture-congruent psychiatric nursing from which guidelines were described. The Nursing for the Whole Person Theory was used as the paradigm. The research method followed three phases--each addressing a different objective. Firstly, explanatory, descriptive research was conducted to compile explanatory case studies reflecting cultural approaches to mental illness and the patients' compliance with psychiatric treatment. The sample consisted of four psychiatric patients purposively selected from four long-term wards of a psychiatric hospital, a group of psychiatric nurses, and the psychiatrists treating each of these patients based on information obtained from the semi-structured interviews, field notes and the patients' clinical documents. From the results of the case studies a practice model for culture-congruent psychiatric nursing was generated using Chinn and Kramer's approach as framework and the major concept "negotiation". Guidelines for a culture-congruent approach in psychiatric nursing were compiled from the model using deductive reasoning and analysis. PMID- 11040632 TI - Health-related problems and proposed solutions identified by women in Ivory Park, Midrand. A participatory research approach. AB - People living in informal settlements do not realise that they could make a significant change in their circumstances by first engaging in introspection and secondly identifying their intellectual resources. In this study, participatory research was done, whereby emphasis was placed on the importance of active participation by all concerned in the project. The participants were women from the community who were able to identify the problems they encountered in their daily living that threatened their health and that of their families. After reflecting on their intellectual resources, an awareness of the power they had over their current situation surfaced. This awareness enabled them to find solutions to their problems. Participatory research is a means of community participation that ultimately leads to empowerment of the community as a whole. PMID- 11040633 TI - Oral health care--the perceptions and self-reported practices of nurses. AB - A structured, 40-item, self-administered questionnaire was distributed to 290 nurses attending an international health care conference held in Pretoria, South Africa. The questionnaire focused on; personal and demographic details of the respondents, history of dental disease and dental attendance, exposure to oral health during and after basic nurse training, opinions on oral health and the nursing curriculum, perceptions on the relationship between nurses and oral health personnel, perceived ability to promote oral health and self-reported contributions to oral health. The 153 respondents were predominantly female (140, 93.3%) with a mean age (+/- Standard deviation) of 43.0 +/- 7.5 years. One hundred and twenty one (79.1%) had previously visited a dental clinic for personal care and 105 (68.6%) had suffered from dental disease, dental pain, bleeding gums or bad breadth. Only 71 (46.4%) were exposed to experiences in assessing and caring for clients with oral disease during their training. Significant positive correlation were found between exposure (during training) to experiences in assessing and caring for clients with oral diseases and some factors. One hundred and forty-eight (96.7%) agreed oral health should be integrated into the nursing curriculum. Forty-one (26.8%) indicated they have not been contributing to the oral health of their clients or community while 92 (60.1%) described their personal contributions as fair. One hundred and thirty six respondents (88.9%) wished to be able to do more for their clients in the area of oral health. PMID- 11040634 TI - [Concept analysis of intuition]. AB - From experience and observation of decision-making in intensive care units, the following has been observed in respect of clinical decision-making. Nurses in intensive care units take clinical decisions they can justify, and other decisions they cannot justify. Nurses in intensive care units sometimes refer to the unjustified clinical decisions as gut-feelings or intuition. The observation regarding clinical decision-making further indicates that clinical decision making, based on intuition is effective and contributes to better and more complete problem-solving capabilities in intensive care units. From the preceding problem statement the following question arose, namely: What is intuition? The objective of the study is to analyze the concept intuition. A philosophical analytical research design was implemented in order to reach this objective. The concept intuition was analyzed by means of concept-analysis. The goal of concept analyses was to respectively describe the characteristics of intuition and developing a theoretical definition for the study regarding these characteristics. From the concept-analysis of intuition the following categories and characteristics of intuition realized namely situational factors of intuition, intuition as a process and intuition as a product. The situational factors of intuition refer to the factors that must be in place in order for intuition to occur, namely knowledge and experience, empathy, as well as incomplete data and uncertain situations. The second group characteristics of intuition has to do with the process of intuition, which is speedy, more synthetical-analogical than analytical, interpretative, holistic, irrational and goal-directed. The third category characteristics of intuition has to do with intuition as a product which include problem-solving and the rational justification of intuition by means of reflection. PMID- 11040635 TI - Health needs assessment of the elderly people of Mangaung. AB - The health needs of the elderly people of Mangaung were assessed through a survey. Three hundred and forty respondents were interviewed. Health needs were clustered around physical, social and psychological aspects. It was found that the elderly cope well in the community and was less hospitalized. There were respondents who were medically diagnosed with physical conditions such as hypertension found in 47.9% and joint diseases in 33.4%, but managed themselves at home. More than 80% had a satisfactory hygiene status although 52.8% could not care for their toenails. Symptoms of depression were found in less than 50% of respondents. With regard to social needs 88.9% had living children who were their source of support. However, more than 40% respondents had no knowledge of community services. This study indicated that the elderly have needs to be met and the nurse can assess and plan for the elderly and refer conditions that needs multidisciplinary approach. PMID- 11040636 TI - Prospective payment from a clinical perspective. PMID- 11040637 TI - The Omnibus Consolidation Appropriations Act of 2000. PMID- 11040638 TI - To salt or not to salt. PMID- 11040639 TI - Take a reality check on telehealth: the nurse is in the picture! PMID- 11040640 TI - PPS--how will it impact your agency and your practice? AB - PPS is being greeted with cautious optimism. The time for change from the cost reimbursed system is long overdue. PPS is intended to reimburse providers by patient need, a system that should provide incentives for quality, cost-effective care. To be fully ready for this change, agencies should prepare for closer tracking of patient care needs and costs, and plan for significant changes in accounting practices. Appropriate staffing patterns and reducing turnover will be important personnel issues. PPS will require nurses to evaluate and plan patient care completely, to make quality visits, to change care in a timely manner when necessary, and to be vigilant in discharge planning and identifying resources. Use of technology, specialty nurses, and closer supervision of care patterns to achieve desired outcomes with maximum efficiency will be important. Clean claims and DDE capability to adjust claims will help ease cash flow problems. Knowing what factors make you a target for medical review will help you to avoid claims delays. Our goal for nursing under PPS seems simple: get our clients well in a timely and cost-effective manner. Seventy-four million baby boomers will enter the Medicare system toward the end of the next decade. Demography, epidemiology, and economics make it clear that a healthy long-term care system is a must. The home health industry's role in that long-term care system will be determined by the home care industry's response to PPS. PMID- 11040641 TI - Improving infection control in home care: from ritual to science-based practice. AB - While many home care policies and procedures are developed with the best of intentions, they frequently lack a scientific basis and have been perpetuated over the years. This article discusses infection control rituals in home care and hospice and the importance of making patient care practice decisions based on sound scientific principles. PMID- 11040642 TI - Are patient information materials too difficult to read? AB - This article describes a study that found much of the written patient information materials used during home visits may be too difficult for patients and caregivers to read and comprehend. A discussion of literacy and suggestions for selecting, developing, and analyzing health education and information materials that are easy-to-read and culturally appropriate are provided. PMID- 11040643 TI - The impact of Medicare reimbursement changes on home healthcare: a nursing perspective. AB - This article presents an overview of changes in Medicare's payment system and their effect on home healthcare from a nursing standpoint, and makes policy recommendations for nurses. Home health care utilization, the Balanced Budget Act 1997, the effect of changes in Medicare reimbursement (on beneficiaries and providers), and transition to the new PPS system are discussed. Suggested strategies agencies can use, and how home care nurses can get involved in policy recommendations are presented. PMID- 11040644 TI - A case study of benefits & potential savings in rural home telemedicine. AB - This case study reports results from a federally funded home telemedicine program in rural Grainger County, Tennessee. Patients, family caregivers, and providers were generally satisfied with this low-cost, user-friendly telemedicine program that used the plain-old-telephone system. Mileage and nurse drive time were reduced and nursing productivity was improved during this demonstration project. PMID- 11040645 TI - Back issues. PMID- 11040646 TI - Home care in the new millennium. PMID- 11040647 TI - In the face of death, nurses are the ultimate healers. PMID- 11040648 TI - Diversity is key to BTN. PMID- 11040649 TI - Embrace lifelong learning. PMID- 11040650 TI - Adam Hohman. Interview by Melissa Ganon. PMID- 11040651 TI - The challenges and rewards of hospice nursing. PMID- 11040652 TI - Palliative care nursing an emerging specialty. PMID- 11040653 TI - When a baby dies. PMID- 11040654 TI - End of life issues and nursing education. PMID- 11040655 TI - A special nurse changes one family's life. PMID- 11040656 TI - Where's the quality? PMID- 11040657 TI - Hope: the antidote to suffering. PMID- 11040658 TI - Pressure ulcer management: the importance of nutrition. AB - Nutrition plays an important role in pressure ulcer prevention and treatment. Nutrition assessment techniques and nutritional interventions for patients at risk for developing a pressure ulcer or who currently have pressure ulcers are essential components of quality patient care. PMID- 11040659 TI - Helping patients with COPD manage episodes of acute shortness of breath. AB - The most disabling and frightening symptom experienced by patients with COPD is dyspnea. Even with the use of bronchodilators, the symptom may not be completely relieved. Patients often develop their own strategies for managing shortness of breath, including the use of a breathing technique called pursed-lip breathing. Although most nurses are familiar with this breathing technique, they often have difficulty assisting patients to use it during acute episodes of shortness of breath. A strategy is described which nurses can use to assist patients in implementing pursed-lip breathing effectively during episodes of acute dyspnea. PMID- 11040660 TI - Pickwickian syndrome: the challenge of severe sleep apnea. AB - Pickwickian syndrome is a severe form of sleep apnea in obese persons which involves mechanical impairment of ventilation resulting in greatly compromised gas exchange. Manifestations of the syndrome are associated with deposits of adipose tissue around the abdomen and diaphragm and are completely reversible with weight loss. Since sleep apnea is now recognized as a significant chronic health problem, nurses in intensive care, medical-surgical, and home care settings are increasingly challenged to provide competent assessment, care, and rehabilitation of affected individuals. PMID- 11040661 TI - Knowing and approaching hope as human experience: implications for the medical surgical nurse. AB - Hospitalized patients frequently share the mystery of their hopes and dreams. Through a deepened understanding and an alternative approach in practice, nurses can come to know patients' hopes and work with them in ways that enhance their quality of life. PMID- 11040662 TI - Caring for patients with coloanal reservoirs for rectal cancer. AB - Advances in surgical techniques enable select patients with rectal cancer to have sphincter-saving procedures that restore the continuity of the GI tract, eliminating the need for a permanent colostomy. One of the preferred surgical options is the construction of a coloanal reservoir or colonic J-pouch. This procedure is usually performed in two stages (two surgeries) and involves creating a temporary ileostomy. Patients undergoing treatment for rectal cancer frequently require adjunctive therapy for the disease before and after surgery. They require extensive education and support during the course of treatment and through rehabilitation. PMID- 11040663 TI - Asthma disease management programs improve clinical and economic outcomes. PMID- 11040664 TI - Using color to simplify ABG interpretation. PMID- 11040665 TI - President's message. A single garment. PMID- 11040666 TI - 2000: the millennium of the student. PMID- 11040667 TI - Strengthening the faculty role: 1951-1971. PMID- 11040668 TI - The future of nursing education. Ten trends to watch. AB - The millennium has become the metaphor for the extraordinary challenges and opportunities available to the nursing profession and to those academic institutions responsible for preparing the next generation of nurses. Signal change is all around us, defining not only what we teach, but also how we teach our students. Transformations taking place in nursing and nursing education have been driven by major socioeconomic factors, as well as by developments in health care delivery and professional issues unique to nursing. Here are 10 trends to watch, described in terms of their impact on nursing education. PMID- 11040669 TI - Recreating nursing practice for a new century. Recommendations and implications of the Pew Health Professions Commission's final report. AB - The erratic path of evolution of health care in the United States creates daunting threats as well as new opportunities for institutions, professionals, and the public. Perhaps no stakeholder has more to risk or gain than professional nursing. As the largest of all the health professions, nursing serves as the backbone for much of the care delivery system. Historically, nurses have been employed primarily in hospital settings. Toward the latter part of the 20th century, however, nurses assumed such expanded roles as independent practitioners, managers of care in large health plans, and providers of alternative and complementary health care services. These new roles point to how nursing may rapidly evolve as the health care system addresses continuing issues of cost, performance, access, and consumer satisfaction. While today's opportunities are great, threats remain to the traditional role of the nurse in the hospital setting, through regulatory constraints on nursing practice and the lack of clear purpose and direction within the broad professional nursing community. Following a decade of leadership and advocacy for health professions education, the Pew Health Professions Commission issued its fourth and final report in December 1998. In this report, the Commission assessed the challenges facing professionals in the 21st century and made general and profession-specific recommendations for action. PMID- 11040670 TI - Health policy and the private sector. New vistas for nursing. AB - During the past two decades, the drive to rein in rising health care costs has shifted some of the power in health care policy making from professional groups, government agencies, and not-for-profit health care organizations to large for profit corporations (1-4). This has been a world-wide phenomenon, as the provision and financing of health care services is shifted from governments to private health care organizations (5,6). In the United States, the shift in power is manifested in profound ways. Market competition and bottom-line economics have permeated the health care system, creating powerful new incentives for mergers, other corporate restructuring, and the shift to for-profit status by formerly not for-profit insurance companies and providers. Private sector health care is now increasingly influenced by for-profit organizations (3). Moreover, the health insurance industry has been transformed as traditional indemnity insurance is replaced by versions of managed care. The role of government, or the public sector, in setting parameters for health care financing and standards for the delivery of health care services is increasingly outpaced in cost cutting by organizations that directly face the bottom line. In addition, private foundations, many of which are under the auspices of managed care organizations, now fund a large proportion of health care research and demonstration projects, a task once largely within the realm of the government. Through education and experience, nurses have developed political sophistication and understanding of policy making in the public sector (7). The challenge now is to educate nurses to adapt their political and policy strategies to the new health care milieu. This challenge is particularly crucial for advanced practice nurses, who must survive in a managed care environment. PMID- 11040671 TI - An inside view. NP/MD perceptions of collaborative practice. AB - The pressing dilemma in this era of health care reform is how to provide cost effective, high quality health care for all Americans. At the present time, due to a number of complex factors, including attrition and economic disincentives, a shortage of primary care physicians exists in certain medically underserved areas of the country. At the same time, however, primary care nurse practitioners are increasing in number. PMID- 11040672 TI - Nursing education and genetics. Miles to go before we sleep. AB - The discoveries of the Human Genome Project (HGP), established in 1990 at the National Institutes of Health and the United States Department of Energy, are bringing important new technologies for genetic diagnosis and treatment to nearly all areas of health care delivery. Nurses, who play an integral role in supporting health consumers as they respond to health and illness, require up-to date genetic knowledge for conducting clinical practice, engaging in nursing research, and educating a new generation of nurses. PMID- 11040673 TI - How does NLNAC support the Pew Health Commission competencies? PMID- 11040674 TI - The other side of illness. PMID- 11040675 TI - Musing on magnets. PMID- 11040676 TI - Patient quality and safety problems in the U.S. health care system: challenges for nursing. PMID- 11040677 TI - Combined benchmarking of hospital outcomes and utilization. AB - A strong case is made for developing quantitative benchmarks for hospital outcomes as well as utilization that includes both acute care re-admissions and lengths of stay. A number of hospitals in two distinctly different geographic health care environments [CA and NY] are studied as to the differences in outcomes and utilization for three of the most common high-cost DRGs. Unscheduled hospital readmissions (within 30 days of initial discharge) were employed as outcome indicators because they reflect both the quality of acute care and the need for case management in the post-discharge period. Benchmark targets were established for patients with a diagnosis of congestive heart failure, acute MI treated medically, or COPD using scattergrams that showed each hospital's mean acute LOS on the x axis and the re-admission rates on the y axis. "Benchmarks were identified as those points with the lowest values for both indicators, as demonstrated by points that were closest to the intersection of the two axes." PMID- 11040678 TI - Expanded role of nursing in ambulatory managed care. Part II: Impact on outcomes of costs, quality, provider and patient satisfaction. AB - This evaluation project used a triangulation of methods and data sources to link the expanded role nurse clinician (NC) to outcomes of costs, quality, and satisfaction in managed care. Results of patient surveys, case studies, cost benefit analyses, interviews, focus groups, and HMO documents suggest that the impact of the role has benefit far beyond its costs. This article builds on an earlier article (Nursing Economic$, Vol. 17, No. 1). Part I reviewed literature on expanded role nursing in population management, and described the development of the NC role. In Part II, the NC expanded role is linked to organizational outcomes of costs, quality, and satisfaction. Results here could be used by managers and planners to provide rationale for implementing expanded nursing roles in managed care in order to improve ambulatory care processes, recruit and retain nurses, reduce costs, and increase patient and provider satisfaction. PMID- 11040679 TI - Redesigning nursing work in long-term care environments. AB - The authors present a highly statistically oriented argument for examining work attitudes and activities among three groups of caregivers [RNs, RPNs, and HCAs] working in long-term care. The investigators used both work sampling, written surveys, and interviews with a sample of 46 caregivers in a large university affiliated LTC facility in Toronto, Canada. While RNs stated their strong affinity for direct patient care activities, they perform the lowest percentage of direct care, chiefly due to their accountability for planning and coordinating the care provided by others. The HCAs who provided the bulk of direct patient care "valued it the least," apparently finding little gratification with this aspect of their role. This study suggests that there is a need to examine and clarify work roles and perceptions for all caregivers as part of any work redesign process. A higher level of RN involvement in direct patient care activities, along with "attention to enhancing the importance" of these activities for staff employed in the HCA role, could be beneficial. PMID- 11040680 TI - Access to health care: an issue central to nursing. PMID- 11040681 TI - Managing effective committees. PMID- 11040682 TI - 'Customerizing' in the new millennium. AB - Many might challenge the concept of customerizing in the health care systems in the face of financial losses caused by the Balanced Budget Act. But, now more than ever, we should be obsessed with the customer and work relentlessly to understand that the procedure is not the end goal. The experience around that procedure will bond the patient to us, or create a consumer who is left with anger. The secret to success, according to Seybold (1998), is "It's the customer, stupid!" (pp. xvi). She also notes that all we have to do is to focus on making it easier for the customer to work with us. We should pick our customers' brains, visit them, learn what they care about, and make it easier for them to work with us. We can't afford not to.$ PMID- 11040683 TI - Redefining the health care landscape with the Internet. PMID- 11040684 TI - Pondering an ethical dilemma. PMID- 11040685 TI - Reaching principled decisions. PMID- 11040686 TI - Living with lymphoedema. PMID- 11040687 TI - Building bridges to clinical practice. PMID- 11040688 TI - In search of an outback nursing adventure. PMID- 11040689 TI - Ensuring the integrity of appraisal systems. PMID- 11040690 TI - Horizontal violence: a challenge for nursing. PMID- 11040691 TI - Communicating the message. PMID- 11040692 TI - New act to bring change. PMID- 11040693 TI - Flexibility now critical. PMID- 11040694 TI - Weight watching. PMID- 11040695 TI - From RADA to Rampton. Interview by Ian McMillan. PMID- 11040696 TI - Floating Hospital. PMID- 11040697 TI - A private life. PMID- 11040698 TI - Making waves. PMID- 11040699 TI - Whisper who dares.... PMID- 11040700 TI - Home alone. PMID- 11040701 TI - Have you seen a ghost? PMID- 11040702 TI - Daytime families. PMID- 11040703 TI - Working solution. PMID- 11040704 TI - Flexible return. PMID- 11040705 TI - ICN on health and human rights. PMID- 11040706 TI - Improving schools' asthma policies and procedures. AB - Collaboration between a health trust and an education authority is helping children with asthma to lead full and active lives at school. Evaluation of the training package, which is delivered by school nurses for school staff, suggests all schools can benefit from a structured approach involving pupils, teachers and parents. PMID- 11040707 TI - Pressure sore care. AB - Christopher Shiels and Brenda Roe report on a survey into the prevalence and care of pressure sores among older people living in residential care and nursing homes in Liverpool. Setting up appropriate information systems and auditing available pressure-relieving equipment are just two of a number of important recommendations arising from the survey. PMID- 11040708 TI - Wound management and pain control. AB - In recent years there have been many advances in the management of wounds, but one area which is still neglected is pain control. The author describes the physiology of wound pain and how careful management can reduce patients' discomfort. PMID- 11040709 TI - Care of the patient with a fractured neck of femur. PMID- 11040710 TI - Take nothing for granted. PMID- 11040711 TI - A test of your skills. PMID- 11040712 TI - Research methods, interventions and evaluation in patient education: a kaleidoscope. PMID- 11040714 TI - Effects of body-mind training and relaxation stretching on persons with chronic toxic encephalopathy. AB - The purpose of this project was to investigate the psychological and physical effects of training of body awareness and slow stretching on persons with chronic toxic encephalopathy (CTE). In the present study, a method of self-regulation, a body-mind training, is presented. The body-mind training used was a guided relaxation technique combined with meditative stretching. The techniques are introduced and the psychological and physiological effects of the training is presented. Eight subjects with CTE, 48.5 years, were trained for 8 weeks. Outcome measures were percentage alpha brain waves (alpha%), electromyography (EMG) on the frontalis muscle, state-trait anxiety (STAI), creativity (RAT), and mood measured as anxiousness, humour and mental fatigue. The mean alpha% increased 52% during the training period (P < 0.01), and the EMG decreased 31% (P < 0.001. State anxiety decreased 22% during the training period (P < 0.01), but no changes were observed in trait anxiety and in the creativity score. The level of anxiousness and fatigue before a training session decreased during the training period. In conclusion, the body-mind training resulted in an improved ability for physical and mental relaxation as indicated from the lower EMG, the higher alpha% and the decrease in state anxiety. PMID- 11040713 TI - Prevalence of passive smoking in infancy in The Netherlands. AB - The objective of the study was to assess the prevalence of passive smoking in infancy. This was done by self-report questionnaires completed by parents who attended the well-baby clinic in the period February-May 1996. A total of 2720 questionnaires were spread among parents with babies between 1 and 14 months: smoking and non-smoking parents. The questionnaires contained questions on smoking habits, smoking at home, smoking in presence of the baby. A total of 1702 parents filled in and returned the questionnaire (63%); 24% of the mothers and 33% of their partners smoked. In 44% of the families, one or more persons smoked; 22% of the mothers and 26% of the partners smoked at home. In 39% of the families, one or both parents smoked at home; 42% of the babies were exposed to tobacco smoke in the living-room, 8% were exposed in the car, and 4% during feeding. In cases where only the mother smoked, 13% of the infants were exposed to tobacco smoke during feeding. In the families where only the partner smoked, the babies were predominantly exposed to smoke in the car (18%). If both parents smoked, the child was most frequently exposed to tobacco smoke in the living-room (73%). It can be concluded that health workers, nurses, pediatricians and family physicians should be advised to inform parents systematically of the harmful effects of passive smoking in infancy. If parents are unable or unwilling to stop smoking, it is important to advise them to refrain from smoking in the presence of the baby. PMID- 11040715 TI - An analysis of the teaching techniques used in diabetic specialist consultations. AB - We attempted to explore the teaching practices of 11 senior diabetic specialists consulting in a out-patient hospital setting and known to have a strong commitment to patient education. The survey consisted of a questionnaire dealing with the duration and distribution of speaking time, the classification of questions, the type of oral questions asked of patients, written materials, teaching aids, demonstrations, if any, care techniques used, and whether or not information was summarized. Our study of 44 consultations showed an average consultation time of 26 +/- 10 min and a balanced sharing of speaking time. The percentage of consultations during which no questions were asked depended on the type and classification of test questions, i.e. oral questions to check knowledge (13.6%), open-ended problem-solving questions (27.3%) or problems involving written materials (54.5%). One to two problems involving open-ended questions were set during 36.4% of the consultations and one to two problems involving written materials were set in 20.5% of the cases; visual aids prepared in advance took a back seat to sketches made during the consultation itself. Information was summed up by the caregiver 75% of the time, and 50% with the patient's help. Our results indicate a patient education orientation during consultations and help to design a standard model formation combined teaching/treatment consultations. PMID- 11040716 TI - Healthy reciprocity in sexual interaction. AB - The purpose of the article is to discuss reciprocity in sexual interaction within a couple relationship in which heterosexual orientation is assumed and satisfaction considered. Reciprocity is modelled as an exchange of services which at its best functions as an unwritten contract, a mutual understanding regarding fairness of returned services, and a desire to comply with this principle together with a loved one/lover. An equal treatment of and balanced attitudes towards one another are present together with a just distribution of benefits and concessions or compromises. Reciprocity involves a relative term although healthy reciprocity can be defined for discussion and assessed as a degree of mutual satisfaction. Sexual interaction issues, skills to obtain satisfaction, and sexual and emotional compatibility are important elements in reciprocity. Understandable communication is an essential contributor in the implementation of reciprocity. Conflict-making dialogue should generally be avoided and connotive meanings of words taken into account. Erotophilia-erotophobia dimensions influence both the learning about and attitudes towards sexuality and contribute to personal and professional abilities to assess sexual problems and to attend to them. Erotic touch is a minimum requirement of love making. Sexual orientation, sexual desire, and intimacy influence sexual compatibility. Equity and exchange models are discussed, and a reciprocity model is proposed. PMID- 11040717 TI - Dutch transmural nurse clinics for chronic patients: a descriptive study. AB - 'Transmural care' can be defined as patient-tailored care provided on the basis of close collaboration and joint responsibility between hospitals and home care organizations. One form of transmural care is transmural nurse clinics for chronically ill. This study describes 62 transmural nurse clinics in the Netherlands. It was established that most of these nurse clinics are held by a specialized community nurse at a hospital outpatient clinic. The principal tasks of the specialized nurse at the clinic are providing illness-related information and supporting patients in dealing with the illness. Only a few unpublished evaluations of Dutch transmural nurse clinics have been conducted. Future research has to provide more insight into the impact of transmural nurse clinics on the quality and continuity of care. PMID- 11040718 TI - Prenatal sickle cell screening education effect on the follow-up rates of infants with sickle cell trait. AB - OBJECTIVE: To assess the effect of prenatal education about newborn sickle cell screening on parents' compliance with the follow-up for infants with sickle cell trait. SUBJECTS: Expectant mothers whose prenatal education included information about newborn sickle cell screening were the study group, and those whose prenatal education did not include such information were the control group. METHODS: Mothers of infants with sickle cell trait were given the opportunity for in-person notification of screening results and follow-up counseling/education. Follow-up rates, anxiety and retention of information were assessed for the case control study. RESULTS: There were a total of 15,670 infants born in the region, and 647 infants were identified with sickle cell trait. The follow-up rate for parents of infants with sickle cell trait was significantly higher (76%) for study group than the control group (49%) (P = 0.0006). Parents whose prenatal education included sickle cell hemoglobinopathy information retained significantly more of the information given during the post-natal education than did controls. CONCLUSION: Our data suggest that prenatal education for expectant mothers which includes information about newborn sickle cell screening significantly increases the follow-up rate for infants with sickle cell trait and contributes to a greater retention of information. PMID- 11040719 TI - Breast cancer patients' experiences of patient-doctor communication: a working relationship. AB - The traumas of diagnosis and treatment for breast cancer are well researched and generally addressed in care. While women with breast cancer continue to identify the need for better communication with physicians, studies to date have not investigated how the process of communication between physicians and women with breast cancer actually unfolds. This phenomenological study therefore explored how women with breast cancer experience patient-physician communication to gain a greater understanding of effective approaches. Interviews of a purposeful sample of 11 women within 6 months of initial diagnosis or recurrence of breast cancer were audiotaped, transcribed verbatim and analyzed using inductive interpretation. Themes and patterns of positive and negative experiences emerged. All experiences began with the woman's feeling of vulnerability. In positive experiences, information sharing and relationship building were inextricably linked components of a working relationship which was at the same time affective, behavioural and instrumental. This experience, in turn, influenced the woman's experience of control and mastery of the illness experience, and their experience of learning to live with breast cancer. Findings illuminate the importance of comprehensively patient-centred, working relationships. Several specific techniques to enhance effective communication are identified. PMID- 11040720 TI - Social support, coping and subjective well-being in patients with rheumatic diseases. AB - The purpose of this cross-sectional study is to examine the relationship between social support, coping, and subjective well-being by testing three hypotheses: (1) social support influences subjective well-being via coping; (2) coping influences subjective well-being via social support; (3) there is a reciprocal relationship between social support and coping, and both concepts influence subjective well-being. Data were analyzed from 628 patients with one or more chronic rheumatic disorder(s) affecting the joints, in some patients combined with another rheumatic disease (no fibromyalgia). Although causal inferences are not possible, the results present a plausible causal sequence in supporting the second hypothesis. This is only true, however, for coping by awaiting/avoidance: coping by awaiting/avoidance led to less social support and this decrease in social support influenced subjective well-being negatively. PMID- 11040721 TI - General practitioners on dementia: tasks, practices and obstacles. AB - The objective of the study was to identify the GPs' perception of their tasks, their practice and obstacles concerning the diagnosis and management of dementia. Twenty-eight GPs participated in focus-group interviews and completed a questionnaire. The GPs perceived their tasks to diagnose, inform and manage dementia patients and their relatives preferably from an early stage on and in such a way that patients are able to stay at home as long as possible. Nevertheless, the GPs diagnose usually in a more progressed stage. As main reasons for this delay the GPs mentioned diagnostic uncertainty during the early stages, embarrassment to conduct a cognitive examination and communicate the diagnosis, non-consulting patients and a lack of time. A discrepancy was found between the GPs' views of their tasks and their clinical practice regarding dementia. Important obstacles were reported that can explain the diagnostic delay and may prevent appropriate education of family caregivers in dealing with demented patients such as embarrassment to examine and communicate this condition. PMID- 11040722 TI - Evaluating health promotion: a tale of three errors. AB - The main purpose of this article is to question the relevance of the Randomised Controlled Trial for the evaluation of health promotion programmes. In its concern to manage Type 1 error, the RCT underestimates or virtually ignores Type 2 and 3 errors. Because of the peculiar complexities of health promotion programmes and the importance of gaining insights into the effect of interventions--rather than merely recording whether or not they achieve their goals--a new kind of validity is needed. The central assertion here is that we should adopt a principle of "judicial review" which is based on a broad spectrum of triangulated evidence. PMID- 11040723 TI - A reliable and valid asthma general knowledge questionnaire useful in the training of asthma educators. AB - Using the responses of 115 adults attending an asthma educator training course, the Asthma General Knowledge Questionnaire for Adults (AGKQA) was found to be an acceptably valid and reliable measure for assessing knowledge related to the management of asthma by adults. Content and face validity: expert assessors considered the AGKQA to be a relevant and plausible test of the asthma general knowledge content of the programme. Criterion-related validity: the pretraining scores of educators were significantly higher (P < 0.001) than those of adults with no experience of asthma; total scores for the AGKQA and an asthma knowledge questionnaire developed for parents of children with asthma correlated strongly, 0.72. Test-retest reproducibility: the Spearman rank correlation for the test retest score was 0.72 (P < 0.02), kappas for concordance of item responses were moderate to very good for two thirds of the items. Internal consistency for the total scale was also acceptable, KR20 0.66. PMID- 11040724 TI - A seven step approach to starting an exercise program for older adults. AB - There is strong experimental evidence to indicate that regular aerobic exercise can prevent disease, decrease the risk of failing, reduce physical disability, improve sleep, and enhance mood and general well being. Despite these benefits, approximately 50% of sedentary adults who start an exercise program stop them within the first 6 months of involvement. To help older adults initiate and adhere to a regular exercise program, a seven step approach was developed and implemented in a continuing care retirement community (CCRC). The seven steps include: (1) education; (2) exercise pre-screening; (3) setting goals; (4) exposure to exercise; (5) role models; (6) verbal encouragement; and (7) verbal reinforcement/rewards. Following implementation of the seven step approach, 40 (19%) of the 212 residents living in the CCRC exercise regularly. PMID- 11040725 TI - Watch, Discover, Think, and Act: a model for patient education program development. AB - In this report we describe the development of the Watch, Discover, Think and Act asthma self-management computer program for inner-city children with asthma. The intervention focused on teaching two categories of behaviors--asthma specific behaviors such as taking preventive medication and self-regulatory processes such as monitoring symptoms and solving asthma problems. These asthma self-management behaviors were then linked with empirical and theoretical determinants such as skills and self-efficacy. We then further used behavioral science theory to develop methods such as role modeling and skill training linked to the determinants. We matched these theoretical methods to practical strategies within the computer simulation and created a culturally competent program for inner-city minority youth. Finally, we planned a program evaluation that linked program impact and outcomes to the theoretical assumptions on which the intervention was based. PMID- 11040726 TI - Watch, Discover, Think, and Act: evaluation of computer-assisted instruction to improve asthma self-management in inner-city children. AB - An interactive multimedia computer game to enhance self-management skills and thereby improve asthma outcomes in inner city children with asthma was evaluated. Subjects aged 6-17 were recruited from four pediatric practices and randomly assigned to the computer intervention condition or to the usual-care comparison. The main character in the game could match the subject on gender and ethnicity. Characteristics of the protagonist's asthma were tailored to be like those of the subject. Subjects played the computer game as part of regular asthma visits. Time between pre- and post-test varied from 4 to 15.6 months (mean, 7.6 months). Analysis of covariance, with pre-test scores, age, and asthma severity as covariates, found that the intervention was associated with fewer hospitalizations, better symptom scores, increased functional status, greater knowledge of asthma management, and better child self-management behavior for those in the intervention condition. Interactions with covariates were found and discussed in terms of variable efficacy of the intervention. PMID- 11040727 TI - Supervised physical activity in Sweden: in theory and practice. AB - The aim of this study was to assess to which extent community-run projects including physical activity could be identified, described and analysed in terms of objectives, organisation, evaluation and financing, as a resource in prevention and treatment of common lifestyle-related medical problems. The Swedish database Spriline was used as a main source of information. Identification of ongoing Swedish activities was followed by a mail questionnaire. In total, 151 projects were eventually identified. A semistructured questionnaire containing about 30 questions was mailed to the individual listed as responsible for the project, with a reminder 2 months later. Only 52 projects were viable; a follow-up of nonresponders showed that no relevant activity program had ever existed or that the person responsible had left. Walking, aerobics and water activities were the dominating types of activity. Most projects addressed both sexes, but eight weight reduction programs were designed for women only. Evaluation ranged from 'measuring attendance' to 'scientific evaluation'. Physical activity programs may not be as systematically organised as the Swedish database suggests and cannot generally be relied upon as support in patient care, unless critically evaluated in advance. PMID- 11040728 TI - Health promotion in community pharmacy: the UK situation. AB - Community pharmacies in the UK are an excellent setting for health promotion in the community; they see over 90% of the population per year. Pharmacy education and training is changing and health promotion is now part of the undergraduate curriculum. There is an increasing body of research about the effects of pharmacy health promotion and a number of examples of good practice. The UK research is reviewed and examples of good practice highlighted. The paper concludes with a discussion about the contribution of pharmacists can make and about some of the issues that will need to be overcome first. Pharmacists see healthy as well as sick people. They have a special relationship with many of their regular customers which enables them to promote health, they can also give population messages to passing trade. A number of issues such as training and remuneration must be addressed before health promotion becomes fully integrated into the pharmacist's role. PMID- 11040729 TI - [Forum Nursing Today 2000: from spring dreams we make marmalade in the fall]. PMID- 11040730 TI - [First consensus-conference in nursing: standards in the prevention of decubitus ulcers]. PMID- 11040731 TI - [Nursing forum Aalen: consequences of the medical product law]. PMID- 11040732 TI - [Forensic patients in general psychiatry--does it work?]. PMID- 11040733 TI - [Administration of a hospital department: guidance and administration has to be learned]. PMID- 11040734 TI - [Kneipp's showers: Prevention is the art of hindering the emergence of a disease]. PMID- 11040735 TI - [Kinesthetics as an instrument for ethical behavior: ethics in nursing needs tools]. PMID- 11040736 TI - [Chaos in life and death: perception is required in order to endure anxiety]. PMID- 11040737 TI - [Power and hierarchy in the world of work: rage in the belly]. PMID- 11040738 TI - [The current problems of German hospitals: the patient is the loser]. PMID- 11040739 TI - [Report from a user: more time for seniors thanks to electronic data processing]. PMID- 11040740 TI - [Introduction of data processing in nursing: report of experiences at the Stuttgart Katharinenhospital]. PMID- 11040741 TI - [Introduction of electronic data processing in nursing: the motivation of the coworkers is a warranty for success]. PMID- 11040742 TI - [Nursing where others are vacationing: first German nursing service on the island of Mallorca]. PMID- 11040744 TI - [The guided self study: an adult form of knowledge transfer]. PMID- 11040743 TI - [Professional diploma course: nursing and health management--aim and profile of a course]. PMID- 11040745 TI - EFM--a house built on sand. PMID- 11040746 TI - Induction of labour for suspected fetal macrosomia. PMID- 11040747 TI - Umbilical cords and underwater birth. PMID- 11040748 TI - Transitional care for neonates. AB - The development of a transitional care ward at Peterborough Maternity Unit has improved the care given to all women and their babies. A slight reorganisation of services has resulted in a better working environment for everyone. Babies requiring additional care receive timely and appropriate attention and mothers whose babies are in NICU are able to support each other at this emotional and stressful time. With such clear benefits for everyone, I recommend that all maternity units consider developing a transitional care ward. PMID- 11040749 TI - Babywatch. Training parents in resuscitation for newborn babies. PMID- 11040750 TI - HIV screening choices. PMID- 11040751 TI - Realigning the service. Evaluating maternity day care in Ninewells Hospital, Dundee. PMID- 11040752 TI - The wall and the ladder. PMID- 11040753 TI - How are you doing? PMID- 11040754 TI - The state of midwifery across England. PMID- 11040755 TI - Acute intrapartum events and cerebral palsy. An international consensus statement. PMID- 11040756 TI - The emotional work of being a midwife. PMID- 11040757 TI - Pleased to meet you. Lesley Page. PMID- 11040758 TI - The needs of the service. PMID- 11040759 TI - Mother love. PMID- 11040760 TI - Health scares: unfair on the public and on health professionals. PMID- 11040761 TI - Can we make child rearing a smoke-free zone? PMID- 11040762 TI - Children's feet: common worries of parents--1. AB - Foot health education, e.g. on correctly fitting socks and shoes, should be part of routine child development checks and health interviews from a very young age onwards. Prevention is better than cure! In-toeing is by far the commonest complaint associated with gait. Many of the complaints raised by parents are simply part of normal development. PMID- 11040763 TI - Menstruation in schoolgirls--1: The normal menarche. AB - The mean age for onset of menstruation (menarche) is 12.8 years. Many schools teach girls about puberty and menstruation and prepare them for the menarche. The school nurse has a valuable part to play in all aspects of menstruation in schoolgirls, psychological as well as physical. The early periods may be irregular and vary markedly in the amount of blood loss. It can take up a year or more for a steady pattern to develop. Dysmenhorrhoea may occur, which may be mild or of the severe spasmodic type. Premenstrual tension syndrome is less common in schoolgirls. Girls should be taught about the importance of hygiene, especially if tampons are used. PMID- 11040764 TI - Impacts of breast-care techniques on prevention of possible postpartum nipple problems. AB - An experimental study was designed to compare the effectiveness of three techniques on the prevention or reduction of nipple pain and cracked nipples during the first ten days postpartum. The methods were to apply warm compresses, to apply expressed breast milk or to give no treatment other than keeping the nipples dry and clean. Applying warm compresses or expressed breast milk was found to be less effective in preventing cracked nipples than simply keeping the nipples dry and clean. The number of cases with nipple pain was less in the group that applied expressed breast milk. PMID- 11040765 TI - A randomised controlled trial of the effects of a cross-cut feeding teat on infant feeding, crying, waking and sleeping behaviour. AB - Feedback from mothers using a "cross-cut" teat indicated its use for problem feeders, particularly "windy" or "colicky" babies. The manufacturer commissioned a trial to see whether this observation was clinically valid. A randomised controlled trial comparing the cross-cut teat with a standard single-hole teat was designed. Both teats were identical in construction apart from the feed hole at the tip of the teat. A standard teat pierced with one hole is usually sold as slow, medium or fast flow rate, depending on the size of the hole. The cross-cut has two slits in the form of a cross opening in response to the baby's sucking. The stronger the suck the wider the cross opens, thus making the flow rate of the teat controlled by the baby. There is therefore no need to change to a teat with a faster flow rate as the baby grows. RESULTS: Babies fed with the cross-cut teat cried less and spent more time awake and content than babies fed with the standard teat. PMID- 11040766 TI - Circumcision: time for honest professionals to speak out against it. PMID- 11040767 TI - The NAS EarlyBird Programme: autism-specific early intervention for parents. PMID- 11040768 TI - [Profile of temporary nurses]. PMID- 11040769 TI - [The temporary nurse: a solution for immediate replacement]. PMID- 11040770 TI - [Temporary nursing: more flexible, more reactive]. PMID- 11040771 TI - [SICS (Substitute Nursing Service): an alternative to temporary nurses]. PMID- 11040772 TI - [Switzerland: an Eldorado for French nurses]. PMID- 11040773 TI - [Industrial nurses: at the center of reform of industrial medicine]. PMID- 11040775 TI - [Industrial nurses. 1. Working at an industrial corporation]. PMID- 11040774 TI - [Inter-university diploma in industrial health care]. PMID- 11040776 TI - [Industrial nurses. 2. Working at a hospital]. PMID- 11040777 TI - Touching. PMID- 11040778 TI - Buying a reliable personal computer. PMID- 11040779 TI - Ethics in action. The family of a comatose, ventilator-dependent patient insists that all forms of treatment be continued. PMID- 11040780 TI - Could you spot this psych emergency? PMID- 11040781 TI - Oncology today: leukemia. PMID- 11040782 TI - Nurse the patient! PMID- 11040783 TI - Advanced practice nursing today. PMID- 11040784 TI - Why we need to worry about warts. PMID- 11040785 TI - Enhanced enteral feeding formulas. PMID- 11040786 TI - Family leave. A Q & A guide. PMID- 11040787 TI - Did a monoclonal antibody cause this patient's death? PMID- 11040788 TI - [Body and languages]. PMID- 11040789 TI - [Body and cultures in Africa]. PMID- 11040790 TI - [The child and its body: the child, its body and sports]. PMID- 11040791 TI - [The child, it's body and sports: a child reaching high levels in sports]. PMID- 11040792 TI - [The child and its body: accidental amputation in children]. PMID- 11040793 TI - [Lively people, pierced bodies, tattooed bodies]. PMID- 11040794 TI - [Mutilate in order to exist?]. PMID- 11040795 TI - [Growth hormone, practices and perspectives]. PMID- 11040796 TI - [Medical strategy: growth hormone, indications, modalities and results]. PMID- 11040797 TI - [Growth hormone, indications, modalities and results. The nurse's role]. PMID- 11040798 TI - [Administration of the hormone and the patient's experience]. PMID- 11040799 TI - [Meeting with Sandrine Turchi, General secretary of the "Growth" association. Interview by Bernadette Fabregas]. PMID- 11040800 TI - [Growth hormone, practice and perspectives: administration of the hormone and the patient's experience, problems and solutions]. PMID- 11040801 TI - [Health and social novelties: pregnancy in adolescence, ways of prevention]. PMID- 11040802 TI - [Impact of the Parental Education Allocation on women's professional activity]. PMID- 11040803 TI - [Rules for a well managed adoption]. PMID- 11040804 TI - [Hospitalization of suicidal adolescents]. PMID- 11040805 TI - [Rude relationships]. PMID- 11040806 TI - [From humiliation to respect for people]. PMID- 11040807 TI - [Ways of addressing each other in a therapeutic relationship]. PMID- 11040808 TI - [Civilities, uncivilities: story of the little spoons]. PMID- 11040809 TI - [Nursing practices: challenge of the first meeting]. PMID- 11040810 TI - [Therapeutic impasse: story of the dragon, known violence, hidden violence]. PMID- 11040811 TI - [Nursing practices elsewhere: care of children of mental patients in the Scandinavian countries]. PMID- 11040813 TI - [From one care to the other: training-education and writing] [In Process Citation] PMID- 11040812 TI - [We have to preserve the framework of the sector politics]. PMID- 11040814 TI - [Pharmacists agree on quality patient information]. PMID- 11040815 TI - [Surfing about nursing care]. PMID- 11040816 TI - [Wounds and bedsores 2000]. PMID- 11040817 TI - [Local treatment of leg ulcers]. PMID- 11040818 TI - [Nutritional status and infection, factors for the delay of cicatrization]. PMID- 11040819 TI - [The "surgical moment" in the care of chronic wounds]. PMID- 11040820 TI - [Use of hemostatic dressings in medical services]. PMID- 11040821 TI - [Decubitus ulcers and the nurses' role]. PMID- 11040822 TI - [Comparative study of 4 kinds of dressings]. PMID- 11040823 TI - [Recommendations for the treatment of decubitus ulcers]. PMID- 11040824 TI - Medical ethics at the dawn of the 21st century. Introduction. PMID- 11040825 TI - Social values, socioeconomic resources, and effectiveness coefficients. An ethical model for statistically based resource allocation. PMID- 11040826 TI - The ethical professor of medicine. Challenges for the twenty-first century. PMID- 11040827 TI - Open heart [Shiva M'Hodu]. PMID- 11040828 TI - Truth telling. PMID- 11040829 TI - The evolution of a hospital ethics committee. PMID- 11040830 TI - Developments in abortion laws. Comparative and international perspectives. PMID- 11040831 TI - The continuing conflict between sanctity of life and quality of life. From abortion to medically assisted death. PMID- 11040832 TI - Advance directives and dementia. PMID- 11040833 TI - The autonomy turn in physician-assisted suicide. PMID- 11040834 TI - A circumscribed plea for voluntary physician-assisted suicide. PMID- 11040835 TI - Peter Singer's theories and their reception in Germany. PMID- 11040836 TI - Problems involved in the moral justification of medical assistance in dying. Coming to terms with euthanasia and physician-assisted suicide. PMID- 11040837 TI - Euthanasia. Reflections on the Dutch discussion. PMID- 11040838 TI - Jurisprudence in the age of biotechnology. An Israeli case analysis. PMID- 11040839 TI - Reproductive liberty and the right to clone human beings. PMID- 11040840 TI - Clones, genes, and reproductive autonomy. The ethics of human cloning. PMID- 11040841 TI - Genetic research. Conversation across cultures. PMID- 11040842 TI - Organ transplantation without brain death. PMID- 11040843 TI - Genetic testing, organ transplantation, and an end to nondirective counseling. PMID- 11040844 TI - Richard Kuhn--Nobel Prize for work on carotenoids and vitamins. PMID- 11040845 TI - Reperfusion for acute myocardial infarction: is the future in plastics? PMID- 11040846 TI - Decreasing mortality with primary percutaneous coronary intervention in patients with acute myocardial infarction: the Mayo Clinic experience from 1991 through 1997. AB - OBJECTIVE: To characterize and determine the overall impact of changes in primary percutaneous coronary intervention (PCI) on the clinical outcome of patients presenting within 24 hours of acute myocardial infarction (AMI). PATIENTS AND METHODS: We retrospectively analyzed a prospective PCI registry for 1073 consecutive patients undergoing primary PCI for AMI at the Mayo Clinic in Rochester, Minn, from 1991 through 1997. The primary outcome measure was mortality from any cause within 30 days and 1 year. RESULTS: The number of patients treated for AMI by primary PCI per year increased from 119 in 1991 to 193 in 1997. Intracoronary stent use increased from 1.7% in 1991 to 64.8% in 1997 (P < .001). This coincided with an increase in ticlopidine use from 3.6% in 1994 to 62.1% in 1997 (P < .001) and in abciximab use from 2.7% in 1995 to 63.2% in 1997 (P < .001). An increase in beta-blocker (58.3% to 75.3%; P < .001), angiotensin-converting enzyme inhibitor (0.9% to 40.0%; P < .001), and 3-hydroxy 3-methylglutaryl coenzyme A reductase use (1.9% to 40.5%; P < .001) as well as a decrease in calcium channel antagonist (34.3% to 8.4%; P < .001) use occurred on discharge. From 1991 through 1997, there was a significant decrease in the 30-day mortality rate (10.1% to 5.2%; P = .05). The 1-year mortality rate also decreased (13.4% in 1991 to 10.4% in 1997) (P = .09). After adjustment for other confounding variables, treatment in more recent years was associated with a significant decrease in death at 30 days (odds ratio, 0.89; 95% confidence interval, 0.79-1.00; P = .05) and during long-term follow-up (odds ratio, 0.93; 95% confidence interval, 0.87-1.00; P = .04). CONCLUSIONS: Percutaneous coronary intervention methods of reperfusion for AMI, along with adjuvant pharmacotherapy, have changed over recent years and have been associated with improved short- and long-term survival. PMID- 11040847 TI - Test-retest reproducibility of the exercise treadmill examination in lumbar spinal stenosis. AB - OBJECTIVE: To provide further validation of the treadmill test by assessing its "test-retest" reproducibility. PATIENTS AND METHODS: In this prospective study, 28 patients with severe lumbar spinal stenosis underwent exercise treadmill testing, first at a walking speed of 1.2 mph and then at the patient's preferred walking speed. All patients had a second treadmill examination or "retest." No treatment intervention was performed between the initial test and the retest. Time to first symptoms (TFS) and total ambulation time (TAT) were measured. Differences between the baseline examination and the retest examination were assessed by using the concordance correlation coefficient (CCC) as well as graphically. RESULTS: There was good reproducibility between baseline test and retest results for all 4 end points: 1.2 mph, TFS (CCC = 0.90); 1.2 mph, TAT (CCC = 0.89); preferred walking speed, TFS (CCC = 0.98); and preferred walking speed, TAT (CCC = 0.96). The median difference between trials was not significantly different from zero for any of the 4 outcomes. CONCLUSIONS: Exercise treadmill testing has good test-retest reproducibility. There was no learning phenomenon associated with the test procedure. The study further validates the clinical utility of exercise treadmill testing in patients with lumbar spinal stenosis and neurogenic claudication. PMID- 11040848 TI - Infective endocarditis in patients receiving long-term hemodialysis. AB - OBJECTIVE: To ascertain the predominant characteristics of patients receiving long-term dialysis who develop infective endocarditis (IE). PATIENTS AND METHODS: We reviewed the records of all chronic hemodialysis patients who had IE at Mayo Clinic, Rochester, Minn, between 1983 and 1997. RESULTS: Twenty episodes of IE occurred in 17 patients. One patient had 3 episodes of IE, and 1 patient had 2 episodes of IE; each episode was caused by a different organism. The mean +/- SD age of our patients was 63 +/- 11 years; there were 13 males; 6 patients had diabetes mellitus; and the mean +/- SD duration of hemodialysis prior to IE was 24.2 +/- 20.5 months. This analysis included 10 episodes of IE (occurring in 9 patients) within the Mayo Clinic Dialysis System during which time 223,358 hemodialysis treatments were delivered, giving a rate of 10 IE episode per 223,336 hemodialysis treatments. Among all 20 IE episodes, there were 14 synthetic arteriovenous grafts, 4 permanent venous dialysis catheters, 2 temporary venous dialysis catheters, and 2 native arteriovenous fistulas (2 accesses in 2 patients), and access had been in place for a mean +/- SD of 15.9 +/- 18.6 months. The portal of infection was the hemodialysis access in 13 episodes of IE. The causative organisms for IE were Staphylococcus aureus in 8 cases, Enterococcus sp in 4 cases, viridans streptococcus in 3 cases, Staphylococcus epidermidis in 2 cases, and 1 case each of Streptococcus bovis, group G beta-hemolytic streptococcus, and Aspergillus sp. The mitral valve was involved in 9 cases, the aortic valve was involved in 5 cases, and the tricuspid and pulmonic valves were involved in 1 case each. Patient survival (after the first episode of IE) was 71% at 30 days; 53% at 60 days; and 35% at 1 year. Echocardiography was performed in 19 episodes of IE. The transthoracic echocardiogram was 62.5% sensitive and 40% specific for the presence of definite or probable vegetations. Univariate analysis for factors affecting 60-day survival show that presence of right-sided IE, vegetation size greater than 2.0 cm3, diagnosis of diabetes mellitus, and initial leukocyte count greater than 12.5 x 10(9)/L were poor prognostic factors. Aortic valve involvement carried a better prognosis. CONCLUSIONS: Infective endocarditis in hemodialysis patients is relatively infrequent but has a high mortality. Patients with synthetic intravascular dialysis angioaccess (synthetic grafts and venous catheters) are more likely to develop IE than patients with native arteriovenous fistulas. Transesophageal echocardiography is a preferred echocardiographic study for suspected cases of IE. Prolonged antibiotic therapy is needed for all patients, and close monitoring is needed for patients with right-sided IE, large vegetations, diabetes mellitus, and an elevated leukocyte count. PMID- 11040849 TI - Anorectal dysfunction in constipated women with anorexia nervosa. AB - OBJECTIVE: To evaluate anorectal and colonic function in a group of patients with anorexia nervosa complaining of chronic constipation. PATIENTS AND METHODS: Twelve women (age range, 19-29 years) meeting the criteria for anorexia nervosa and complaining of chronic constipation were recruited for the study. A group of 12 healthy women served as controls. Colonic transit time was measured by a radiopaque marker technique. Anorectal manometry and a test of rectal sensation were carried out with use of standard techniques to measure pelvic floor dysfunction. A subgroup of 8 patients was retested after an adequate refeeding program was completed. RESULTS: Eight (66.7%) of 12 patients with anorexia nervosa had slow colonic transit times, while 5 (41.7%) had pelvic floor dysfunction. Colonic transit time normalized in the 8 patients who completed the 4-week refeeding program. However, pelvic floor dysfunction did not normalize in these patients. CONCLUSIONS: Patients with anorexia nervosa who complain of constipation have anorectal motor abnormalities. Delayed colonic transit time is probably due to abnormal eating behavior. PMID- 11040850 TI - Management and extended outcome of patients with synchronous bilateral solid renal neoplasms in the absence of von Hippel-Lindau disease. AB - OBJECTIVE: To gain information regarding long-term follow-up in patients with synchronous bilateral solid renal neoplasms in whom renal-preserving surgery is imperative. PATIENTS AND METHODS: We examined our surgical experience and the survival outcome, as evaluated by Kaplan-Meier and log-rank analysis, of 94 patients (64 men and 30 women) who presented to the Mayo Clinic in Rochester, Minn, from 1973 to 1998 with bilateral synchronous solid renal neoplasms in the absence of von Hippel-Lindau disease. Follow-up of these patients ranged from 1 to 25 years, with a mean of 5.86 years and a median of 4.18 years. Tumors were staged according to the TNM classification. Pathologic staging and grading were usually performed on the kidney with the most extensive cancer. The Cox proportional hazards model was used to assess the relationship of grade (1-4), tumor size, and enucleation as opposed to extended (1 cm) partial nephrectomy on overall, cancer-specific, local recurrence-free, and metastasis-free survival. RESULTS: Seventy-one patients (76%) had bilateral synchronous renal cell carcinoma, and 14 patients (15%) had a unilateral renal cell carcinoma with a contralateral benign solid neoplasm. Nine patients (10%) had bilateral benign solid lesions. Sixty-six patients (70%) underwent a single procedure, whereas 28 (30%) underwent staged surgical procedures. Fifty-one patients (54%) are alive, and 43 (46%) have died. Twenty patients (21%) died of metastatic disease, and 5 (5%) had a local recurrence. Cancer-specific survival of the 85 patients with at least 1 renal cell carcinoma still under observation was 81% (+/- 4.9% SE) and 59% (+/- 8.1% SE) at 5 and 10 years, respectively, and survival to local recurrence was 96% (+/- 2.6% SE) at 5 years and 93% (+/- 3.7% SE) at 10 years with 14 patients still under observation. Grade 3 was a statistically significant factor for metastasis (P < .001). A significant difference in metastasis-free survival and cancer-specific survival was noted dependent on pathologic T stage (P < .001 and P = .02, respectively), with patients with local pT3 disease having a higher rate of metastasis and cancer-specific death. Multivariate analysis revealed that tumor grade was associated with metastasis-free survival (P = .002) and tumor size with cancer-specific survival (P = .04). There was no statistical significance on survival outcome end points according to procedure performed, i.e., enucleation vs extended partial nephrectomy. CONCLUSION: Long-term results of renal-preserving procedures for a series of patients with bilateral solid renal neoplasms indicate that grade, stage, and tumor size are significant predictors of outcome. Mean follow-up of over 5 years supports nephron-sparing techniques in selected patients because local recurrence was infrequent compared with distant metastasis. PMID- 11040851 TI - The "aspirin" of the new millennium: cyclooxygenase-2 inhibitors. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) inhibit prostaglandin synthesis via the cyclooxygenase (COX) enzyme, the key to both therapeutic benefits and toxicity. COX enzyme exists in 2 isoforms, COX-1 and COX-2. COX-1 enzyme is thought to mediate "housekeeping" or homeostatic functions, and COX-2 is considered an inducible enzyme in response to injury or inflammation. COX-2 inhibitors are the "next-generation" NSAIDs that may selectively block the COX-2 isoenzyme without affecting COX-1 function. This may result in control of pain and inflammation with a lower rate of adverse effects compared with older nonselective NSAIDs. Rapidly evolving evidence suggests that COX-2 enzyme has a diverse physiologic and pathologic role. This article addresses the role of COX-2 enzyme in health and disease as well as the potential therapeutic value and safety issues related to COX-2 inhibition. PMID- 11040852 TI - Infection and immunity in chronic lymphocytic leukemia. AB - Patients having chronic lymphocytic leukemia (CLL) are at increased risk for infectious morbidity and mortality. The predisposition to infections in CLL patients has many components, including both immunodeficiency related to the leukemia itself (humoral and cellular immune dysfunction) and the results of cumulative immunosuppression related to CLL treatment. The risk of infectious complications increases with the duration of CLL, reflecting the natural history of the disease and the cumulative immunosuppression related to its treatment. Hence, in early, untreated CLL, the infectious risk is mainly related to hypogammaglobulinemia, and infections by encapsulated bacteria are common. However, in patients having advanced CLL, particularly those who receive the newer purine analogues, neutropenia and defects in cell-mediated immunity appear to be the major predisposing factors. An expanded spectrum of pathogens, including opportunistic fungi, Pneumocystis carinii, Listeria monocytogenes, mycobacteria, and herpesviruses, are seen in that setting. The changing spectrum of infections in this latter group of patients mandates a newer approach to prophylaxis and therapy. PMID- 11040853 TI - Prevalence, pathophysiology, and treatment of patients with asthma and gastroesophageal reflux disease. AB - About one third of the US adult population experiences symptoms of gastroesophageal reflux on a monthly basis. Asthma is present in about 5% of the same population. This article reviews and summarizes the literature in the following areas: (1) prevalence of gastroesophageal reflux disease (GERD) in asthmatic patients based on clinical symptoms, endoscopic esophagitis, and 24 hour ambulatory esophageal pH recordings; (2) proposed pathophysiologic mechanisms linking the 2 diseases; and (3) medical and surgical treatment trial results of antireflux therapy for asthmatic patients. Asthmatic patients appear to have an increased prevalence of GERD symptoms and 24-hour esophageal acid exposure. The clinical management of these patients remains controversial. Common management approaches to GERD in asthmatic patients include medical therapy with a proton pump inhibitor and/or antireflux surgery, which improve asthma symptoms in many patients but minimally affect pulmonary function. PMID- 11040854 TI - The ethical validity and clinical experience of palliative sedation. AB - The physician's main goal in caring for a dying person is to reduce suffering, including pain, physical symptoms, and emotional, psychosocial, and spiritual distress. In refractory and intractable cases, palliative sedation offers a compassionate and humane alternative to conscious and continual suffering, both for the patient and the patient's family. Without a doubt, further studies are necessary, particularly in cases of cognitive impairment, but palliative sedation offers a valuable and efficacious intervention for interminable suffering at the end of life. PMID- 11040855 TI - Hypertension in pregnancy: diagnosis and treatment. AB - Hypertension affects 10% of pregnancies in the United States and remains a leading cause of both maternal and fetal morbidity and mortality. Hypertension in pregnancy includes a spectrum of conditions, most notably preeclampsia, a form of hypertension unique to pregnancy that occurs de novo or superimposed on chronic hypertension. Risks to the fetus include premature delivery, growth retardation, and death. The only definitive treatment of preeclampsia is delivery. Treatment of severe hypertension is necessary to prevent cerebrovascular, cardiac, and renal complications in the mother. The 2 other forms of hypertension, chronic and transient hypertension, usually have more benign courses. Optimal treatment of high blood pressure in pregnancy requires consideration of several aspects unique to gestational cardiovascular physiology. The major goal is to prevent maternal complications without compromising uteroplacental perfusion and fetal circulation. Before an antihypertensive agent is prescribed, the potential risk to the fetus from intrauterine drug exposure should be carefully reviewed. PMID- 11040856 TI - 70-year-old woman with chest pain and new diastolic murmur 6 months after coronary artery bypass grafting. PMID- 11040857 TI - Calcified bicuspid aortic valve mass prolapsing into the left main coronary artery. AB - We report a case of a mobile calcific mass on the aortic valve that prolapsed into the left main coronary artery of a 51-year-old man. This case and a review of the literature suggest that calcific embolization to coronary arteries is a rare but possibly underrecognized complication of calcified degenerative or bicuspid aortic valves. This potentially catastrophic complication of calcified aortic valves needs to be suspected and recognized in clinical practice. PMID- 11040858 TI - Superficial temporal artery to middle cerebral artery bypass and external carotid reconstruction for carotid restenosis after angioplasty and stent placement. AB - The recent proliferation of endovascular treatment of carotid atherosclerotic disease will increase the number of patients who require treatment for recurrent carotid stenosis after angioplasty and stent placement. The optimal management of these patients has not yet been defined. We describe a 66-year-old woman who required 2 surgical procedures for recurrent in-stent carotid stenosis. She experienced numerous transient ischemic attacks 5 months after left extracranial internal carotid artery angioplasty and stenting for asymptomatic stenosis. Angiography showed high-grade in-stent restenosis, left intracranial carotid artery stenosis, and poor collateral flow to the left middle cerebral artery circulation. The patient underwent a superficial temporal artery to middle cerebral artery bypass, and the transient ischemic attacks resolved. Five months later, angiography showed progressive stenosis of the external carotid artery at the site of the stent. The patient underwent successful external carotid reconstruction with an on-lay patch. Extracranial-intracranial bypass grafting may be used successfully in the treatment of recurrent extracranial carotid artery stenosis after angioplasty and stent placement. Also, external carotid artery reconstruction at the site of an internal carotid artery stent can be performed safely. PMID- 11040859 TI - Etanercept for the treatment of human immunodeficiency virus-associated psoriatic arthritis. AB - Etanercept may play an important role in modulating the inflammatory activity and progression of human immunodeficiency virus (HIV)-associated psoriasis and psoriatic arthritis. We report the case of a 45-year-old homosexual man with a CD4 cell count of less than 0.05 x 10(9)/L and an HIV viral load of 4200 copies/mL (while receiving highly active antiretroviral therapy) who developed extensive psoriatic plaques, 4.5-kg weight loss, onychodystrophy, and psoriatic arthropathy with severe periarticular bone demineralization. The arthritis progressed despite the use of several disease-modifying medications, including corticosteroids, hydroxychloroquine, and minocycline. Because of uncontrolled, progressive, and disabling arthritis and resulting profound disability, he was treated with etanercept. Within 3 weeks, his psoriasis had improved dramatically and his joint inflammation had stabilized. For the next 4 months, immunologic and viral parameters remained stable, but his clinical course was complicated by frequent polymicrobial infections. Etanercept was thus discontinued despite continued improvements in his psoriasis, psoriatic arthritis, and functional status. While both cutaneous and joint manifestations of psoriasis improved dramatically, the experience with this patient dictates that caution and careful follow-up must be exercised when prescribing etanercept in the setting of HIV infection. PMID- 11040860 TI - Paradoxical embolization in an adult patient with cystic fibrosis. AB - To our knowledge, we describe the first reported case of paradoxical embolization via a patent foramen ovale (PFO) in an adult with moderately severe cystic fibrosis (CF) and advanced lung disease. Fluctuating neurologic symptoms and signs suggestive of cerebrovascular disease in an adult patient with advanced CF may be due to paradoxical embolization via a PFO. The possibility of a PFO should be considered before placement of a totally implantable venous access device to avert unnecessary risk of stroke in CF patients. Further study is needed to determine whether the use of a totally implantable venous access device increases the risk of paradoxical embolization in adult CF patients with a PFO. PMID- 11040861 TI - New antifungal agents that inhibit the growth of Candida species: dichlorinated 8 quinolinols. AB - Five dichlorinated 8-quinolinols (2,5- 5,6-, 3,5-, 3,7-, and 4,5-dichloro-8 quinolinol) were tested against Candida albicans and C. tropicalis in Sabouraud dextrose broth with and without bovine serum. The 5,6-, 3,5-, and 3,7-dichloro-8 quinolinols proved to be more effective than the control, 5-fluorocytosine. In cytotoxicity tests employing baby hamster kidney (BHK) cells, all test agents proved to be more cytotoxic than the control. However, the minimum inhibitory concentration (MIC) of 3,5-dichloro-8-quinolinol to both fungi was only one tenth the cytotoxic dose, suggesting that the compound may be useful as a topical or systemic antifungal agent. PMID- 11040862 TI - Cryptococcus neoformans in bird excreta in the city zoo of Cali, Colombia. AB - The presence of Cryptococcus neoformans was studied in bird excreta and in the air circulating in and around bird cages in the City Zoo of Cali, Colombia, between August 1994 and April 1995, using a sunflower seed agar culture medium for fungus isolation. A total of 380 samples was studied, 110 from droppings and 270 from Petri dishes placed inside (148) and outside (122) the cages. C. neoformans var neoformans was found in only two cases, one from bird excreta (0.9%) and the other from air inside a cage (0.7%). The former positive sample was collected from the cracks of a dead tree where two crested caracaras (Polyborus plancus) roosted; the feces were dry, accumulated, and with a pH of 6. The other positive sample was found inside the cage of these birds; however, samples taken in a dispersion study at 0.5, 1, 5 and 10 m around this cage were all negative. It appears that this low isolation rate is due to adequate cleaning and disinfection procedures used in the city zoo of Cali. PMID- 11040863 TI - Antifungal activity of some marine organisms from India, against food spoilage Aspergillus strains. AB - Crude aqueous methanol extracts obtained from 31 species of various marine organisms (including floral and faunal), were screened for their antifungal activity against food poisoning strains of Aspergillus. Seventeen species exhibited mild (+ = zone of inhibition 1-2 mm) to significant (+3 = zone of inhibition 3-5 mm) activity against one or the other strain under experiment. However, extracts of 12 species were active against all the three strains. Organisms like Salicornia brachiata (obligate halophyte), Sinularia leptocladus (Soft coral), Elysia grandifolia (Mollusks), Gorgonian sp. 2 and Haliclona sp. exhibited significant (inhibition zone of 3-5 mm) antifungal activity against one or the other strains. However, extracts of A. ilicifolius, Amphiroa sp., Poryphyra sp., Unidentified sponge, Suberites vestigium, Sinularia compressa, Sinularia sp., Sinularia maxima, Subergorgia suberosa, Echinogorgia pseudorassopo and Sabellaria cementifera were mild (inhibition zone of 1-2 mm) to moderate (inhibition zone of 2-3 mm) active against the respective strains. The growth of A. japonicus was significantly inhibited by the extracts of S. leptocladus (r = 0.992, p < 0.0001) and E. grandifolia (r = 0.989, p < 0.0001). PMID- 11040864 TI - Effect of climatic conditions on natural mycoflora and fumonisins in freshly harvested corn of the State of Parana, Brazil. AB - Natural mycoflora associated with fumonisins were analyzed in 150 samples of freshly harvested corn from Central-Southern, Central-Western and Northern regions of the State of Parana, Brazil and correlated to climatic conditions. The corn samples were frequently contaminated with Fusarium sp. (98.7 to 100%) and Penicillium sp. (93 to 100%), when compared to Aspergillus sp. (not detected to 27.7%). The highest contamination with potentially mycotoxigenic fungi occurred in corn harvested in the Central-Western region, where total mould and yeast counts ranged from 5.5 x 10(3) to 5.2 x 10(6) CFU/g, with 98.7% contaminated by Fusarium sp. and 93% by Penicillium sp. In this region F. moniliforme (F. verticillioides) was the predominant Fusarium sp., and was isolated in 85.9% of the samples. Aspergillus sp. was isolated from 27.7% samples. FB1 was detected in 100% of the samples (mean of 2.39 micrograms/g) and FB2 in 97.7% (mean of 1.09 micrograms/g). Fumonisins were also detected in all samples from Northern region, with mean of 4.56 micrograms/g (FB1) and 2.20 micrograms/g (FB2). Considering 1.0 microgram/g as the threshold, 72% of the corn samples from the Central-West and 92% from the North were contaminated with concentrations above this value, in contrast to a 18.5% contamination rate from Central-Southern samples. Between corn planting to harvesting season, the average maximum temperature and relative humidity were 26 degrees C and 77.1% (Central-Southern), 27 degrees C and 69% (Northern) and 29.9 degrees C and 89.1% (Central-Western). Therefore, the higher fumonisins contamination of corn from Northern region when compared to the Central-South were due to the differences in rainfall levels (92.8 mm in Central Southern, 202 mm in Northern) during the month preceding harvest. PMID- 11040866 TI - The occurrence of HT-2 toxin and other trichothecenes in Norwegian cereals. AB - A total of 449 grain samples, 102 barley, 169 wheat and 178 oat samples were collected from different regions of Norway from 1996-1998 crops, mainly from grain loads and silos. The samples were analysed for type A and B trichothecenes, the largest groups of mycotoxins produced by the Fusarium species, by gas chromatography with mass spectrometric detection (GC-MS). Factors affecting the presence of the different trichothecenes are discussed. Deoxynivalenol (DON) and HT-2 toxin were the trichothecenes most frequently detected, followed by T-2 toxin, nivalenol, and scirpentriol, scirpentriol being detected only in seven samples (> 20 micrograms/kg). Oats were the grain species most heavily contaminated with an incidence (% > 20 micrograms/kg) and mean concentration of positive samples of 70% (115 micrograms/kg) for HT-2 toxin, 30% (60 micrograms/kg) for T-2 toxin, 57% (104 micrograms/kg) for DON, and 10% (56 micrograms/kg) for nivalenol. The corresponding values for barley were 22% (73 micrograms/kg), 5% (85 micrograms/kg), 17% (155 micrograms/kg) and 6% (30 micrograms/kg), and for wheat 1.2% (20 micrograms/kg), 0.6% (20 micrograms/kg), 14% (53 micrograms/kg) and 0% for HT-2, T-2, DON and nivalenol, respectively. Norwegian oats were found to contain HT-2 and T-2 toxin in concentrations that might be at threat to human health for high consumers of oats. The amount of DON was significantly lower than in the crop from previous years. PMID- 11040865 TI - Cytotoxicity of four trichothecenes evaluated by three colorimetric bioassays. AB - The application of cell culture technique for screening of low concentrations of Fusarium mycotoxins was examined. Three colorimetric bioassays were used to determine the cytotoxicity of the trichothecenes T-2 toxin (T-2), HT-2 toxin (HT 2), deoxynivalenol (DON) and nivalenol (NIV) to 3T3 mouse fibroblasts (3T3 cells). The bioassays assess DNA synthesis (incorporation of 5-bromo-2' deoxyuridine; BrdU), metabolic activity (cleavage of 3-(4,5-dimethyltiazol-2-yl) 2,5-diphenyltetrazolium bromide; MTT) and cell membrane damage (release of lactate dehydrogenase; LDH), respectively. The BrdU bioassay was the most sensitive and the IC50 values (50% response compared to untreated cells) of T-2, HT-2, DON and NIV were 4.6, 13, 263 and 365 ng/ml, respectively. At the same toxin concentrations used in the BrdU bioassay, only T-2 and HT-2 were toxic enough to obtain IC50 values using the MTT bioassay. The IC50 values for T-2 and HT-2 were 12 and 68 ng/ml, respectively. When determined by the LDH bioassay, the IC50 values of T-2 and HT-2 were 18 and 42 ng/ml, respectively. At the tested concentrations, DON and NIV had a minor effect on the 3T3 cells when evaluated by the MTT and LDH bioassays. The BrdU bioassay in combination with 3T3 cells was found to be a suitable method for determination of trichothecene-induced toxicity at low concentrations. PMID- 11040867 TI - Physiologic and behavioral assessment of rabbits immunized with Freund's complete adjuvant. AB - Although the use of Freund's Complete Adjuvant (FCA) has been discouraged for the production of polyclonal antibodies, little clinical evidence supports the belief that FCA necessarily affects the well-being of immunized rabbits. We designed the present study to determine whether immunization at multiple sites with small volumes of Freund's adjuvant affects rabbit well-being. We injected 18 female New Zealand White rabbits (six animals per group) with antigen in FCA, Freund's Incomplete Adjuvant, or physiologic saline in the following volumes and routes: 0.02 to 0.03 mL intradermally in each of 30 to 40 sites and 0.1 mL subcutaneously in each of two sites. The body weight, temperature, complete blood count, and behavior of the rabbits in the home cage, upon handling, and in an open field did not differ significantly among the immunization groups during the 7-week assessment period. Only the degree of induration around injection sites differed: as expected, FCA induced the greatest response at the injection sites, but the sites were neither ulcerative nor necrotic, nor did palpation of the sites induce any apparent discomfort to the rabbits. We conclude that FCA may be used safely and humanely in rabbits if small volumes are injected intradermally or subcutaneously in multiple sites. PMID- 11040868 TI - Cardiovascular parameters telemetrically measured during pregnancy, parturition, and lactation in a common marmoset (Callithrix jacchus). AB - Here we present the first report of heart rate, blood pressure, and locomotive activity of a female common marmoset (Callithrix jacchus) that was moving freely in its home cage during gestation, parturition, and lactation. We collected the data by using a miniaturized telemetry and data-acquisition system (Data Sciences, St. Paul, MN) that had been adapted for marmosets (1). The parameters were recorded continually from 12 days before to 15 days after parturition. Parturition lasted about 3.5 h, during which marked changes in heart rate, systolic blood pressure, and diastolic blood pressure occurred. To obtain control values, we again measured these parameters for 13 days during a "physiologically neutral" phase (4 months after parturition), when the female was neither pregnant nor lactating. Heart rate was 200% higher, systolic blood pressure was 25% higher, and diastolic blood pressure was 75% higher during parturition than during the neutral phase. PMID- 11040869 TI - Urolithiasis in cynomolgus monkey (Macaca fascicularis): a case report. AB - Only a few cases of urinary calculi have been reported in cynomolgus monkeys. The adult male Macaca fascicularis presented here had two calculi, both of which were 100% calcium carbonate. Similar calculi occur in humans. PMID- 11040870 TI - Reproducible echocardiography in juvenile sheep and its application in the evaluation of a pulmonary valve homograft implant. AB - Increased use of the ovine animal model in cardiovascular surgical research has created a salient need for standardized echocardiography techniques. To demonstrate a reproducible image in this species and confirm the validity of echocardiography as a diagnostic tool, we implanted 10 sheep with a pulmonary valve homograft and monitored them through weekly echocardiographic examinations until 20 weeks after implantation. We obtained good images from the left cranial and the left caudal transducer windows without needing to sedate the animals. Sedated sheep yielded adequate views from the right apical window. Echocardiographic data on the implanted homografts (including functional capacity, presence of calcification, and hemodynamic information and measurements), completely agreed with the results of the post-explantation examinations. PMID- 11040871 TI - Method and complications of ileocutaneous anastomosis for collection of ileal digesta in neonatal pigs. AB - The purpose of our study was to develop a surgical method for collection of ileal digesta in neonatal (< 5 kg) pigs and to determine potential complications of the procedure. In 18 10-day-old pigs, we performed ileocutaneous anastomosis (ICA) via a right ventrolateral incision. The ICA was readily performed in these neonatal pigs; one pig died 24 hours after surgery because of intestinal volvulus. Pigs were monitored twice daily for development of post-operative complications. Ileal digesta were collected "free-catch" by using metabolism cages because attempts to use cannulas (diameter, 4 to 8 mm) and collection bags failed. To determine the effect of colon bypass on hydration, electrolytes, glucose, and serum enzyme activities, we collected serum biochemistry data before and 6 days after surgery. Changes in serum biochemical values included increased potassium, creatinine, total protein, albumin, and globulin and decreased ALP and glucose, but all values remained within normal ranges for neonatal pigs. ICA is tolerated well by neonatal pigs and is an easily learned and rapid technique for collection of ileal digesta. In addition, ICA is a useful alternative to "T cannulas" and ileorectal anastomosis for nutrition research using neonatal pigs weighing < 5 kg. PMID- 11040872 TI - Preparation of the inguinal fat pad for perfusion in situ in the rat: a surgical technique that preserves continuous blood flow. AB - We developed a surgical technique for preparing the inguinal fat pad of rats for perfusion that preserves continuous blood flow to the tissue. Fatty acid uptake from the fat pads of fed rats (46.1 +/- 2.1% of arterial supply, n = 82) and release from those of fasted (48 h) animals (51.3 +/- 3.1% of supply, n = 69) occurred principally via the free fatty acid component of the blood; levels of triglycerides, cholestryl esters, and phospholipids did not change significantly. Venous blood flow in perfusions using blood from fed rats was 83.3 +/- 1.7 mL/min and 83.1 +/- 1.5 microL/min in experiments involving fasted donors. Values for arterial (A) and venous (V) pH (A, 7.41 +/- 0.03; V, 7.32 +/- 0.04), pO2 (A, 151.6 +/- 15.7 mm Hg; V, 34.7 +/- 9.2 mm Hg), pCO2 (A, 30.8 +/- 6.6 mm Hg; V, 58.2 +/- 5.2 mm Hg), and hematocrit (A, 44.6 +/- 1.2%; V, 45.7 +/- 1.2%) were unchanged throughout the course of the perfusions. Fat pad and blood perfusates were maintained at 37 degrees C. Tissue homogenates revealed that the total fatty acid content of fat pads from fed rats (333.5 +/- 0.3 mg/g tissue) differed significantly from that in fasted animals (260.7 +/- 0.7 mg/g tissue; P < 0.001). Our technique likely will have many uses in the study of lipid transport and metabolism, hyperlipidemia, and cancer-associated cachexia. PMID- 11040873 TI - Congenital bilateral ureteral stenosis and hydronephrosis in a neonatal puppy. AB - Three days after an uneventful parturition, a Brittany spaniel/beagle puppy (Canis familiaris) was nursing but not gaining weight as rapidly as were its littermates. Although its diet was supplemented, the puppy died 10 days after birth. The renal pelves were enlarged and filled with urine. Both ureters were thin throughout their length, and urine could not be expressed from either kidney into its respective ureter. The bladder contained no urine and was firmly embedded in the umbilicus. Histologically, both kidneys were hydronephrotic and contained hypoplastic collecting tubules. The diameter of the right (0.55 mm) and left (0.57 mm) ureters at the uteropelvic junction were narrower than those of an age-matched control of the same breed (1.03 mm and 1.02 mm) and were lined by hypoplastic urothelium. Trichrome staining of the ureters revealed excessive collagen and disorganized smooth muscle fibers; in contrast, the control had predominantly circular smooth muscle fibers and less fibrous tissue. Although neither blood nor aqueous humor could be evaluated for urea nitrogen, we suspect that the puppy died from uremia. The congenital bilateral ureteral stenosis and hydronephrosis of the described puppy is similar to a form of uteropelvic obstruction in humans. PMID- 11040874 TI - Novel use of the Elizabethan collar as a "hoop-skirt" for protecting wounds and catheters in nonhuman primates. AB - This article describes the novel use of an Elizabethan collar, which is attached to a primate jacket to create a tamper-proof "hoop-skirt" for protecting wounds and catheters. We successfully have used this hoop-skirt to manage juvenile male rhesus monkeys with Foley catheters for 10 days post-prostatectomy. In addition, our hoop-skirt has been used to manage wounds on the hindlimbs of both macaques and baboons. PMID- 11040875 TI - Corneal dermoid in a dwarf rabbit (Oryctolagus cuniculi). AB - Histology after superficial keratectomy verified our diagnosis of corneal dermoid in a dwarf rabbit (Oryctolagus cuniculi). To our knowledge, this lesion has not been reported previously to occur in dwarf rabbits. Ocular dermoids should be considered as a differential diagnosis for corneal abnormalities in the rabbit. PMID- 11040876 TI - Fluorogenic 5' nuclease PCR (real time PCR). PMID- 11040877 TI - Reform of the Mental Health Act. Health or safety? PMID- 11040878 TI - Developmental psychiatry--insights from learning disability. AB - BACKGROUND: The Blake Marsh lecture, an annual lecture on learning disability, was endowed in 1963 in memory of Dr Blake Marsh, the former medical superintendent of Bromham House Colony in Bedford. The first lecture was given in 1967. AIMS: To review the speciality of the psychiatry of learning disability and how it is currently practised in the UK. METHOD: Clinical, service, research and educational issues in learning disability psychiatry are reviewed and illustrated. RESULTS: Key issues which emerge in all four areas include the importance of communication skills, consultation with users and carers, professional education and partnership. CONCLUSIONS: The psychiatry of learning disability is a complex, varied and stimulating branch of psychiatry with a strong developmental focus. PMID- 11040879 TI - West London first-episode study of schizophrenia. Clinical correlates of duration of untreated psychosis. AB - BACKGROUND: Studies in schizophrenia suggest that a longer initial period of untreated illness is associated with a poorer clinical outcome. AIMS: To determine whether, in first-episode schizophrenia, a longer duration of untreated psychosis (DUP) or of untreated illness (DUI) (DUP plus any prodrome) is associated with clinical variables that could mediate a poor prognosis. METHOD: Clinical, social, neuropsychological and oculomotor function data on 53 patients with first-episode schizophrenia were related to the DUP and DUI. RESULTS: Comparing short and long DUP groups split around the median showed no statistically significant differences (except age); patients in the latter group tended to perform worse on an executive attentional set-shifting task, and were more likely to be unemployed, and living alone or homeless. CONCLUSIONS: There was little evidence of any association between either DUP or DUI and progressive deterioration in the schizophrenic illness or the development of resistance to initial drug treatment. Social variables that augur a poor prognosis may be associated with delayed presentation of schizophrenia to psychiatric services. PMID- 11040880 TI - Causes of the excess mortality of schizophrenia. AB - BACKGROUND: The excess mortality of schizophrenia is well recognised, but its precise causes are not well understood. AIMS: To measure the standardised mortality ratio (SMR) and examine the reasons for any excess mortality in a community cohort with schizophrenia. METHOD: We carried out a 13-year follow-up of 370 patients with schizophrenia, identifying those who died and their circumstances. RESULTS: Ninety-six per cent of the cohort was traced. There were 79 deaths. The SMRs for all causes (298), for natural (232) and for unnatural causes (1273), were significantly higher than those to be expected in the general population, as were the SMRs for disease of the circulatory, digestive, endocrine, nervous and respiratory systems, suicide and undetermined death. Smoking-related fatal disease was more prominent than in the general population. CONCLUSIONS: Some of the excess mortality of schizophrenia could be lessened by reducing patients' smoking and exposure to other environmental risk factors and by improving the management of medical disease, mood disturbance and psychosis. PMID- 11040881 TI - No association between breast-feeding and adult psychosis in two national birth cohorts. AB - BACKGROUND: It has been proposed that breast-feeding might have a protective effect against the development of adult schizophrenia. AIMS: To test this hypothesis. METHOD: Using prospective data from two UK national birth cohorts, the feeding histories of those who later developed schizophrenia were compared with the remaining population at risk. Analyses in each cohort were considered to be independent tests of the hypothesis. RESULTS: There were no differences in feeding histories. In the 1946 birth cohort (n = 4447) 30 cases of DSM-III-R schizophrenia arose by age 43; 24.1% of cases v. 23.6% of controls were entirely bottle-fed; 17.3% v. 12.3% were breast-fed for under 1 month; 58.6% v. 64.1% were breast-fed beyond 1 month. In the 1958 cohort (n = 18,856), 40 cases of CATEGO nuclear schizophrenia arose by age 28; 24.1% of cases v. 31.7% of controls were entirely bottle-fed; 27.6% v. 24.9% were breast-fed for under 1 month; 48.3% v. 43.4% were breast-fed beyond 1 month. CONCLUSIONS: These findings provide no evidence of any effect of breast-feeding in protecting against the risk of later schizophrenia. PMID- 11040882 TI - Changes in regional cerebral blood flow due to cognitive activation among patients with schizophrenia. AB - BACKGROUND: The regional cerebral blood flow (rCBF) response to the Wisconsin Card Sort Test (WCST) has been used to assess the functional integrity of the prefrontal cortex in patients with schizophrenia. AIMS: In this study, patients were divided into two groups according to whether they had made few or many perseverative errors on a modified version of the WCST. A control group consisted of normal volunteers. The groups were then compared with respect to rCBF response to WCST activation. METHOD: rCBF was measured during administration of a modified version of the WCST and during a card sorting control task, using single photon emission computerised tomography (SPECT). RESULTS: Performance of the modified WCST was associated with a widespread and substantial increase in rCBF, particularly in the frontal region. The poorly performing group of patients with schizophrenia showed only a modest increase in rCBF in the left anterior cingulate region. CONCLUSION: Subjects with schizophrenia are able to respond to specific neuropsychological challenge with activation of the frontal regions. PMID- 11040883 TI - The revised Beliefs About Voices Questionnaire (BAVQ-R). AB - BACKGROUND: We present a revised Beliefs About Voices Questionnaire (BAVQ-R), a self-report measure of patients' beliefs, emotions and behaviour about auditory hallucinations. AIMS: To improve measurement of omnipotence, a pivotal concept in understanding auditory hallucinations, and elucidate links between beliefs about voices, anxiety and depression. METHODS: Seventy-one participants with chronic auditory hallucinations completed the BAVQ-R, and 58 also completed the Hospital Anxiety and Depression Scale. RESULTS: The mean Cronbach's alpha for the five sub scales was 0.86 (range 0.74-0.88). The study supports hypotheses about links between beliefs, emotions and behaviour, and presents original data on how these relate to the new omnipotence sub-scale. Original data are also presented on connections with anxiety and depression. CONCLUSIONS: The BAVQ-R is more reliable and sensitive to individual differences than the original version, and reliably measures omnipotence. PMID- 11040884 TI - Reappraising insight in psychosis. Multi-scale longitudinal study. AB - BACKGROUND: Many patients suffering from psychosis are unaware of their disorder and symptoms. AIMS: To investigate whether insight changes with time, and how it relates to patients' psychopathology, and to examine the correlations between insight scales in patients with psychoses. METHODS: Seventy-five consecutively admitted in-patients with schizophrenia, affective disorder with psychotic symptoms, or schizoaffective disorder were examined after remission of an acute episode and at follow-up (> 6 months). Three different scales were used to assess insight. RESULTS: To some extent, insight into past episodes improved over time in patients with psychosis, regardless of diagnosis. Few significant relationships between insight and psychopathology remained stable at follow-up. The higher the negative and disorganisation dimensions at baseline, the less did attitudes to treatment vary when tested at follow-up. No predictive value for variability of psychopathological dimensions was found for insight dimensions. The insight scales used were highly intercorrelated, suggesting that they measure the same construct. CONCLUSIONS: Insight and psychopathology seem to be semi independent domains. PMID- 11040885 TI - Ethnic differences in admissions to secure forensic psychiatry services. AB - BACKGROUND: Persons of African-Caribbean origin are more frequently imprisoned, and increasing evidence suggests they are detained more frequently in psychiatric hospitals, following offending behaviour. AIMS: To estimate population-based prevalence rates of treated mental disorder in different ethnic groups compulsorily admitted to secure forensic psychiatry services. METHOD: A survey was recorded of 3155 first admissions, from 1988 to 1994, from half of England and Wales, with 1991 census data as the denominator adjusted for under enumeration. RESULTS: Compulsory admissions for Black males were 5.6 (CI 5.1-6.3) times as high as, and for Asian males were half, those for White males; for Black females, 2.9 (CI 2.4-4.6) times as high and for Asian females one-third of those for White females. Admissions of non-Whites rose over the study period. Patterns of offending and diagnoses differed between ethnic groups. CONCLUSIONS: Variations in compulsory hospitalisation cannot be entirely attributed to racial bias. Community-based services may be less effective in preventing escalating criminal and dangerous behaviour associated with mental illness in African Caribbeans. PMID- 11040886 TI - Substance misuse as a marker of vulnerability among male prisoners on remand. AB - BACKGROUND: More treatment for substance misuse should be provided within prisons. AIMS: To examine differences between prisoners on remand with substance misuse problems and other prisoners on remand. METHOD: Random selection and interview of unconvicted male prisoners (n = 750, a 9.4% sample), plus examination of the prison medical record. RESULTS: Of the sample of 750, 253 subjects (33.7%) reported either drug- or alcohol-related health problems or dependency. Compared with other prisoners on remand, they reported more childhood adversity, conduct disorder, self-harm, past psychiatric treatment and current mood disorder, and had fewer qualifications, were more likely to be unemployed and have more housing difficulties. CONCLUSIONS: One-third of unconvicted men in prison report substance-related problems, and these are a marker for vulnerability within a disadvantaged population. Health care providers should involve this group in treatment and rehabilitation, both inside prison and following release. PMID- 11040887 TI - The Clinician Assessment of Fluctuation and the One Day Fluctuation Assessment Scale. Two methods to assess fluctuating confusion in dementia. AB - BACKGROUND: The identification of fluctuating confusion is central to improving the differential diagnosis of the common dementias. AIMS: To determine the value of two rating scales to measure fluctuating confusion. METHOD: The agreement between the clinician-rated scale and the scale completed by a non-clinician was determined. Correlations between the two scales were calculated; variability in attention was calculated on a computerised cognitive assessment and variability in delta rhythm on an electroencephalogram (EEG). RESULTS: The Clinician Assessment of Fluctuation and the computerised cognitive assessment were completed for 155 patients (61 Alzheimer's disease, 37 dementia with Lewy bodies, 22 vascular dementia, 35 elderly controls). A subgroup (n = 40) received a further evaluation using the One Day Fluctuation Assessment Scale and an EEG. The two scales correlated significantly with each other, and with the neuropsychological and electrophysiological measures of fluctuation. CONCLUSIONS: Both scales are useful instruments for the clinical assessment of fluctuation in dementia. PMID- 11040888 TI - Brain weight in suicide. An exploratory study. AB - BACKGROUND: There is little available literature on the effect of suicide methods on brain weight. AIMS: To explore variations in post-mortem brain weight in different methods of fatal self-harm (FSH) and in deaths from natural causes. METHOD: A review of a sample of coroners' records of elderly persons (60 and above). Verdicts of suicide, misadventure and open verdicts were classified as FSH. Post-mortem brain weight for 142 FSH victims and 150 victims of unexpected, sudden or unexplained death due to natural causes, and from various methods of FSH, were compared. RESULTS: Brain weight of victims of FSH was significantly higher than of those who died of natural causes (P < 0.01); brain weights in both groups were within the normal range for this age group. There was no significant difference in brain weight between different methods of FSH (P > 0.05). CONCLUSIONS: The findings require critical examination and further research, to include data from younger age groups. A regional or national suicide neuropathological database could be set up if all victims of FSH underwent routine neurohistochemical post-mortem examination. PMID- 11040889 TI - Price, cost and value of opiate detoxification treatments. Reanalysis of data from two randomised trials. AB - BACKGROUND: Treatments in different settings have different costs. A dilemma arises if expensive treatments lead to better outcomes. AIMS: To investigate conflicts between the priorities of cost minimisation, clinical effectiveness, and cost-effectiveness in the detoxification of opiate addicts. METHOD: Cost and clinical effectiveness were examined using published outcome data. The main outcome measures were: achieving a drug-free state on completion of detoxification; the economic costs of treatment. RESULTS: In terms of simple cost, in-patient detoxification is much more expensive than out-patient treatment (ratio, 24:1). With adjustment for successful outcome, the costs are almost identical (ratio, 0.9:1). Comparison of specialist and general psychiatry in patient settings showed that even when adjusted for clinical outcomes, the specialist setting is more costly (ratio, 1.9:1), although the outcomes are better. CONCLUSIONS: Naive adherence to cost and cost-containment considerations is dangerous. Discussion of treatment costs is misleading if not informed by, and adjusted for, evidence of effectiveness. This is especially important where marked differences in outcome between treatment options exist. PMID- 11040890 TI - Expenditure on mental health care by English health authorities: a potential cause of inequity. AB - BACKGROUND: The York resource allocation formula includes a calculation of the amount needed to purchase mental health services equitably in each health authority in England. However, the amount which is actually spent on services is at the discretion of the authority. AIMS: To compare expenditure on mental health services with allocation, and test the hypothesis that differences between them are to the disadvantage of services in deprived areas. METHOD: A comparison of routine expenditure and allocation data, and linear regression modelling of the ratio of expenditure to allocation. RESULTS: The ratio of expenditure to allocation varies widely. Relative underspending occurs more frequently in deprived areas, although not in the four inner-London health authorities. CONCLUSIONS: The intentions of the York formula are not achieved in practice. The implications of the formula for mental health should be made explicit to health authorities, and shortfalls in mental health expenditure relative to allocation should be justified at a local level. PMID- 11040891 TI - Relationship between psychotic disorders in adolescence and criminally violent behaviour. A retrospective examination. AB - BACKGROUND: The interaction between psychosis and violence in adults is an important area of research receiving attention. To date there is little available data examining this relationship in adolescence. AIMS: To investigate the possible relationships between criminally violent types of behaviour, and psychopathology and social factors, among adolescents suffering from a psychotic disorder. METHOD: A retrospective case note study of 39 in-patients diagnosed as having a psychotic disorder and admitted to one of two adolescent psychiatry units (one secure, one open). Cases were divided into a 'violent' and a 'non violent' group, and these two groups were then compared for social and psychopathological variables. RESULTS: There was no association between recorded psychopathology and criminally violent behaviour. Criminally violent behaviour was associated with a history of emotional or physical abuse, contact with social or mental health services, and previous criminal behaviour. CONCLUSIONS: These findings fail to echo results of studies in adult schizophrenia; they suggest that violent behaviour in psychosis is associated more closely with social factors than with specific symptoms of the psychotic illness. Potential explanations are discussed. PMID- 11040892 TI - Fear reduction by psychotherapies: a response. PMID- 11040893 TI - Psychological debriefing: historical military perspective. PMID- 11040894 TI - Psychosocial treatment programmes for personality disorders: service developments and research. PMID- 11040895 TI - More disappointing treatment outcomes in late-life depression. PMID- 11040896 TI - Medication and alcohol in nursing homes. PMID- 11040897 TI - Talmudic, Koranic and other classic reports of stalking. PMID- 11040899 TI - Challenges in ophthalmic pathology: the vitreoretinal membrane biopsy. AB - The introduction of vitreoretinal microsurgery has produced a new type of biopsy; that of the vitreoretinal membrane. This review investigates methods by which these scar-like tissues are handled in the laboratory and explores the implications of the results of such evaluations. The study of vitreoretinal membrane biopsies has provided much information concerning the pathobiology of the various conditions which may give rise to the tissue as well as insights into how membranes themselves develop. Moreover, the application of new laboratory techniques is expected to enhance our understanding of the formation of vitreoretinal membranes, and lead to further advances in their surgical and medical management. PMID- 11040898 TI - Venlafaxine-induced painful ejaculation. PMID- 11040900 TI - Anterior ischaemic optic neuropathy associated with central retinal vein occlusion. AB - PURPOSE: To report the unusual association between non-arteritic anterior ischaemic optic neuropathy (NAION) and non-ischaemic central retinal vein occlusion (CRVO) in two patients. METHODS: Case reports are presented. RESULTS: Non-ischaemic CRVO was manifested by dilated, tortuous retinal veins with flame shape retinal haemorrhages. Fluorescein angiography showed prolonged arteriovenous transit time and normal retinal capillary perfusion without macular oedema. The presence of colour vision abnormalities, relative afferent pupillary defects, pale disc swelling and visual field deficits indicated that the visual loss was attributable entirely to NAION. Laboratory investigations disclosed impaired fibrinolytic function in case 1 and the presence of antiphospholipid antibodies in case 2. CONCLUSIONS: Compression of the central retinal vein by the swollen optic nerve could have predisposed to CRVO. The presence of thrombophilic abnormalities may have contributed to the concomitant occlusion of posterior ciliary arteries and central retinal vein. Ischaemic optic neuropathy needs to be considered in patients with CRVO when the visual acuity is not consistent with the retinal pathology. PMID- 11040901 TI - Detection of retinal lesions after telemedicine transmission of digital images. AB - PURPOSE/BACKGROUND: To assess whether loss of image resolution or colour and subsequent telemedicine transmission of digital images affects the accuracy of retinal lesion detection by ophthalmologists when compared with the original transparencies. METHODS: Fifteen ophthalmologists of different experience independently scored 11 retinal images for pathological signs. The images were presented as either transparencies or colour and monochrome digital images, which had been transmitted via telephone lines to a geographically remote location. One patient's eye was also imaged using scanning laser ophthalmosocopy (SLO) which produced a dynamic black and white digital image. ANOVA analysis was performed. RESULTS: Total scores were higher for transparencies than colour (p = 0.0003) or black and white digital images (p = 0.00006). Expert observers (n = 5) considered separately showed no significant difference of accuracy between transparencies and either colour digital (p = 0.09) or monochrome digital images (p = 0.11). Experts were better than trainees at detecting pathology from less familiar images: total score (p = 0.02), colour digital (p = 0.03), monochrome digital (p = 0.02) and SLO images (p = 0.004). CONCLUSION: Experienced observers can identify sight-threatening retinal pathology from poorer-resolution digital images that have been transmitted by telemedicine. They can also adapt to viewing less familiar images such as black and white digital or SLO images. PMID- 11040902 TI - Who should manage primary retinal detachments? AB - PURPOSE: To determine whether the outcome of primary retinal reattachment surgery in a subregion is improved by surgery being performed in a specialist vitreoretinal unit (VRU). METHODS: A subregional, population-based, retrospective audit cycle of primary retinal reattachment surgery was conducted by independent investigators. The subregion was defined as the catchment area of a teaching hospital (TH) with a specialist VRU and three neighbouring district general hospitals (DGHs). During the initial audit period (January 1989 to December 1990), 142 cases were treated at all four hospitals: TH/VRU (83), DGH-A (15), DGH B (13), and DGH-C (31). Policy changes after the initial audit led to primary retinal reattachment surgery being predominantly performed by the VRU. During the re-audit period (September 1995 to August 1997), 160 cases were treated at two hospitals: VRU (148) and DGH-C (12). The outcome measure employed was complete retinal reattachment after a single procedure with a minimum follow-up of 12 months. RESULTS: The success rate for primary retinal reattachment surgery in the subregion improved from 76.1% to 88.8% (p = 0.006) following the policy changes. The success rate of the vitreoretinal specialists in the VRU (90%) was greater than the general ophthalmologists in the DGHs (ranging from 47% to 77%), despite case selection by the general ophthalmologists. The number of cases treated by the VRU increased by 156% in the 6.5 year interval between the two audits due to a widespread change in the model of care for primary retinal detachments (both within and outside the subregion). During the re-audit period, the VRU treated 348 primary retinal detachments (including referrals from outside the subregion), achieving a success rate of 86.8% with a single procedure and 97.4% with further surgery. This primary success rate included 35 cases (10%) treated by vitrectomy with silicone oil tamponade who did not undergo silicone oil removal. CONCLUSIONS: The outcome of primary retinal reattachment surgery can be improved if surgery is performed by a specialist VRU. It is suggested that the current standard for retinal reattachment with a single procedure should be set in the region of 85% to 90%. Changing the model of care so that primary retinal reattachment surgery is predominantly performed by a specialist VRU has important resource implications. PMID- 11040903 TI - Screening for diabetic retinopathy using digital colour photography and oral fluorescein angiography. AB - PURPOSE: To evaluate digital colour photography and oral fluorescein angiography (OFA) for diabetic retinopathy screening. METHODS: Thirty-seven patients were selected from either a diabetic retinopathy screening or a medical retina clinic. Three 45 degrees colour digital images and a single macula 45 degrees OFA image were taken from each eye. Standard seven-field stereo photography with ETDRS grading was used as a gold standard for data comparison. The images were assessed by two graders and the results of each method compared using the McNemar test. RESULTS: Five eyes had no diabetic retinopathy, 50 had background diabetic retinopathy, 3 had pre-proliferative diabetic retinopathy, 11 had proliferative disease and 3 had quiescent posttreatment disease. Clinically significant macular oedema was present in 25 eyes and absent in 48. For grading diabetic retinopathy digital colour photography produced a sensitivity of 0.87 (specificity 0.83); OFA produced a sensitivity of 0.87 (specificity 0.80) (p = 0.1). For the detection of diabetic maculopathy, the sensitivity of digital colour photography was 0.48 (specificity of 0.95) and for OFA was 0.87 (specificity 0.87) (p < 0.01). CONCLUSION: This pilot study has shown that both digital colour photography and OFA compare well with conventional methods for diabetic retinopathy screening. The results encourage the further evaluation of OFA in the screening for diabetic maculopathy. PMID- 11040904 TI - Changes in retinal light sensitivity following blunt ocular trauma. AB - PURPOSE: To prospectively examine changes in retinal light sensitivity following blunt ocular trauma in eyes with traumatic hyphaema with no evidence of retinal injuries 1 week, 1 month and 4 months after injury. METHOD: Sixteen patients who sustained hyphaema after blunt ocular trauma without visible traumatic retinal lesions underwent visual field testing on the C 30-2 programme of the Humphrey field analyser as soon as visual acuity recovered to 0.7 or more with correction. RESULTS: Significant reduction in retinal sensitivity with MD (mean deviation) p values less than 5% was found in 50% (8 eyes) of our patients 1 week after injury. MD p values significantly improved 1 month after injury (p < 0.001) and even more so after 4 months (p < 0.00004). After 4 months only in one eye (6%) was a significant reduction in retinal sensitivity observed with a MD p value less than 5%. No correlations between improvement in retinal sensitivity and the extent of hyphaema or the extent of angle recession were observed. CONCLUSIONS: In 50% of eyes following blunt ocular trauma without evident traumatic posterior segment abnormalities significant transient reduction in retinal light sensitivity was observed with nearly complete recovery over time. PMID- 11040905 TI - Plasma total homocysteine and retinal vascular disease. AB - PURPOSE: Hyperhomocysteinaemia has been linked to macrovascular disease. Our aim was to investigate whether there is a relationship between fasting plasma total homocysteine levels and retinal vascular disease. METHODS: We measured the homocysteine levels in 70 patients with arterial or venous retinal vessel occlusion and compared them with the levels in 85 controls without evidence of ischaemic heart disease. Homocysteine levels were determined by high-performance liquid chromatography with electrochemical detection and compared after logarithmic transformation. RESULTS: Homocysteine levels were found by univariate analysis (unpaired two-tailed t-test) to be significantly higher in the group with retinal artery occlusion than the group with retinal vein occlusion (p = 0.045) and in both groups compared with controls (18.4 and 13.8 vs 9.5 mumol/l; p = 0.0002 and < 0.0001, respectively). The controls, however, were significantly younger than the subjects (51.5 +/- 15.4 vs 66.2 +/- 11.9 years; p < 0.0001), but analysis of the results by age revealed significant differences between the groups and controls for the seventh decade (vein occlusions, p = 0.05) and for the eighth decade (artery occlusions, p = 0.037). Subgroup analysis of the retinal vessel occlusion group revealed significant differences in mean blood pressure between those with branch retinal vein occlusions (175/100 mmHg) and both those with central retinal vein occlusions (155/88 mmHg) and those with retinal artery occlusions (157/86 mmHg). Both vein occlusion subgroups also differed significantly with regard to homocysteine levels, branch < central (12.2 +/- 1.3 vs 15.0 +/- 1.6 mumol/l, p = 0.03). Multiple linear regression analysis revealed significant relationships between homocysteine levels and the presence of retinal vessel occlusion (p = 0.0002), serum creatinine (p = 0.001) and age (p = 0.003), but not gender. CONCLUSIONS: We conclude that homocysteine may be a risk factor for retinal vascular disease and could be simply and cheaply treated with folate and vitamins B6 and B12. PMID- 11040906 TI - Prilocaine versus lignocaine for minor lid procedures. AB - PURPOSE: To determine whether prilocaine is a more comfortable local infiltration anaesthetic agent than the more widely used lignocaine for minor eyelid procedures. METHODS: A prospective randomised study was undertaken to compare the discomfort between local infiltration of plain 2% prilocaine versus its equivalent, plain 2% lignocaine. One hundred and twenty-five patients were recruited. Pain was assessed subjectively using a visual analogue pain score, graded from 0 to 10. RESULTS: The mean pain score for the prilocaine group was 1.82 compared with 3.19 for the lignocaine group. Using the Mann-Whitney U-test for significance, U = 1236.5; p < 0.001. CONCLUSION: Prilocaine is a more comfortable local infiltration anaesthetic agent than lignocaine when used for minor eyelid procedures. PMID- 11040907 TI - The glabellar flap dissected. AB - The glabellar flap is effective in the reconstruction of defects within the medial canthal region. The unique contour of the medial canthal region presents a challenge in the surgical planning of the glabellar flap, which is only briefly described in texts. We describe its step by step construction, including the indications and precautions. We also include a section on variations in design for improved closure. With careful planning, the glabellar flap provides excellent cosmetic results. PMID- 11040908 TI - Erdheim-Chester disease: two cases of orbital involvement. AB - Erdheim-Chester disease (ECD) is an increasingly recognised form of fibro inflammatory process characterised by xanthomatous histiocytes containing large amounts of ingested lipid, plasma cells, macrophages and Touton-type giant cells. Ophthalmic involvement in ECD has been reported in only 22 cases. We describe two patients, one presenting with diabetes insipidus and subsequently developing orbital pseudotumours and retroperitoneal fibrosis, the other presenting with exophthalmos and diplopia. The first patient was treated with cladribine and subsequently developed sudden onset of bilateral blindness while the second required radiation therapy for the retro-orbital process and developed radiation retinopathy. These cases typify the variable presentation and course in patients with ECD. PMID- 11040909 TI - The rates of blindness and of partial sight registration in glaucoma patients. AB - PURPOSE: To determine the extent of unregistered blind and partial sight visual loss amongst primary open angle glaucoma (POAG) patients in an outpatient clinic. METHODS: A 13 year follow-up study was carried out of all patients with POAG attending the outpatient clinics at the Leicester Royal Infirmary during the first 4 months of 1982. RESULTS: Ninety (35%) of 258 patients achieved eligibility for registration and 47 patients (18%) were registered, consisting of 39 who were eligible and 8 who were not. Fifty-seven per cent of eligible patients remained unregistered. Patients with visual loss due to visual acuity loss were much more likely to be registered than patients with either visual field loss (p < 0.001) or mixed visual acuity/visual field loss (p < 0.001). All categories of eligible patients experienced a delay between eligibility and registration. This delay was much longer for the visual field loss patients (median delay 61.8 months). Patients with untreatable disease were more likely to be registered. The rates of registration are improving. CONCLUSION: A large proportion of glaucoma patients who are eligible for registration as either blind or partially sighted remain unregistered. Those who are registered often experience prolonged delays before becoming registered. PMID- 11040910 TI - Self-reported eye disease in elderly South Asian subjects from an inner city cluster in Bradford: a small-scale study to investigate knowledge and awareness of ocular disease. AB - PURPOSE: To investigate various issues relating to eye diseases in a sample of 200 South Asian residents living in an inner city regional cluster in Bradford. METHODS: Door to door interviews were carried out by one investigator who spoke English, Punjabi and Urdu. Visual acuity and pinhole acuity were measured using a portable LogMAR acuity chart. A structured questionnaire investigated various issues including self-reported eye disease, knowledge of eye disease, the effectiveness of various sources for eye-related information, the importance of early detection of eye diseases and the need for adequate control of systemic diseases linked to eye diseases such as diabetes. RESULTS: Data revealed poor knowledge of self-reported eye diseases and of the importance of early detection and inadequate knowledge of the link between the control of systemic disease such as diabetes and ocular complications. Language barrier problems and poor utilisation of available ocular health care services were also evident. CONCLUSIONS: These data, although not exhaustive, give an insight into various factors that affect the ocular health of the South Asian community in Bradford. PMID- 11040911 TI - Does ethnic origin influence the incidence or severity of keratoconus? AB - PURPOSE: Keratoconus affects all races, yet very little information exists as to the relative frequency in patients of different ethnic origin. We aimed to establish the incidence and severity of keratoconus in Asian and white patients. METHODS: The hospital records of the ophthalmology department of a large Midlands hospital with a catchment population of approximately 900,000 (87% white, 11% Asian, 2% other) were examined retrospectively for the 10 year period from 1989 to 1998. RESULTS: For the age group 10-44 years the prevalence of keratoconus in Asians and whites was 229 and 57 per 100,000 respectively, a relative prevalence of 4 to 1. The incidence of keratoconus in the same age group was 19.6 and 4.5 per 100,000 per year respectively, a relative incidence of 4.4 to 1. Asians were significantly younger at presentation compared with whites (mean 22.3 +/- 6.5 vs 26.5 +/- 8.5 years, p < 0.0001). A first corneal graft was carried out on 14% of the Asian and 15% of the white patients. Of those having grafts, Asians were significantly younger than white patients at the time of diagnosis (mean 19.1 +/- 4.8 vs 25.7 +/- 7.3 years, p = 0.005) and at operation (mean 21.4 +/- 5.0 vs 28.7 +/- 7.7 years, p = 0.004). The interval from diagnosis to operation, though shorter for Asians, was not significantly different (mean 1.8 +/- 1.4 vs 2.5 +/- 1.7 years, p = 0.2). CONCLUSION: The results show previously unrecognised racial differences in the hospital presentation of keratoconus in the UK. Compared with white patients, Asians have a fourfold increase in incidence, are younger at presentation and require corneal grafting at an earlier age. PMID- 11040912 TI - Management of pellucid marginal corneal degeneration. AB - PURPOSE: A retrospective study to ascertain the management of pellucid marginal corneal degeneration (PMCD). METHOD AND RESULTS: Sixteen patients (average age 42.6 years) presented with PMCD. PMCD was bilateral in 13 and unilateral in 3 patients. Eight eyes underwent surgery. Nineteen eyes were managed non surgically. Surgery involved corneal wedge excision (WE) (6 eyes), penetrating keratoplasty (PK) (3 eyes) and lamellar thermo-keratoplasty (LTK) (1 eye). Immediate pre-operative average visual acuity (VA) was 6/24, 6/10 and 6/60 with an average pre-operative astigmatism of 11.40 D, 9.75 D and 20.5 D for WE, PK and LTK respectively. After an average post-operative follow-up of 57 months, 66 months and 1 year, the average astigmatism was 8.90 D, 4.63 D and 6.00 D with an average final VA of 6/19, 6/15 and 6/6 for WE, PK and LTK respectively. In the nonsurgical group, at presentation, 40% of eyes had a VA of 6/12 or better. After an average follow-up period of 32.3 months, 80% of eyes had a visual acuity of 6/12 or better. Optical correction was achieved with spectacles and or contact lenses. CONCLUSIONS: Surgical correction for PMCD provides poor long-term reduction of astigmatism. Patients with PMCD may be adequately corrected in the long term by the use of scleral fitted gas-permeable contact lenses. PMID- 11040913 TI - The role of tear physiology in ocular surface temperature. AB - PURPOSE: To determine whether the more rapid cooling of the tear film in dry eyes is related to other tear film parameters, a battery of tear physiology tests was performed on dry eye patients and control subjects. METHODS: Tear evaporation rate was measured with a modified Servomed (vapour pressure) evaporimeter and ocular temperature with an NEC San-ei 6T62 Thermo Tracer in 9 patients diagnosed as having dry eye and in 13 healthy control subjects. Variability in temperature across the ocular surface was described by the temperature variation factor (TVF). Lipid layer structure and tear film stability were assessed with the Keeler Tearscope and tear osmolality was measured by freezing point depression nanolitre osmometry. RESULTS: The data were explored by principal component analysis. The subjects with and without dry eye could be separated into two distinct groups entirely on the basis of their tear physiology. Dry eye patients exhibited higher tear evaporation rates, osmolalities and TVF, lower tear film stabilities and poorer-quality lipid layers than the control subjects. A significant linear relationship was found to exist between tear evaporation rate and TVF for all subjects (R2 = 0.242, p = 0.024). CONCLUSIONS: Rapid cooling of the tear film in dry eyes appears to be related to the reduced stability of the tears and the increased rate of evaporation. The higher latent heat of vaporisation, associated with the increased evaporation in dry eyes, may account for the increased rate of cooling of the tear film in this condition. PMID- 11040914 TI - The effect of pre-operative topical flurbiprofen or diclofenac on pupil dilatation. AB - PURPOSE: To assess the clinical benefit and relative efficacy of pre-operative diclofenac and flurbiprofen drops in routine cataract surgery. METHODS: Fifty-two patients undergoing extracapsular cataract extraction with lens implantation were randomised in a double-masked study to compare the efficacy of diclofenac, flurbiprofen and placebo drops in maintaining per-operative mydriasis and reducing post-operative inflammation. Balanced salt solution containing adrenaline was used in all patients. Pupil size was measured prior to the corneal section and after the completion of the operation. The degree of pain, redness, flare and cells in the anterior chamber and intraocular pressure were recorded on the day after surgery. The three groups were analysed with respect to change in pupil size, intraocular pressure and degree of inflammation. RESULTS: The change in pupil size was significantly different among the three groups (p = 0.01), there being a smaller decrease in the treatment groups compared with the placebo group and in the diclofenac treatment group compared with the flurbiprofen treatment group. Significantly less post-operative redness was recorded in the diclofenac treatment group compared with the other groups (p = 0.001). No significant difference was found between the groups as regards anterior chamber cells, flare or intraocular pressure change. CONCLUSIONS: Pre-operative diclofenac and flurbiprofen drops are effective in maintaining intraoperative mydriasis. Diclofenac reduces post-operative redness on day 1. These effects are of debatable clinical benefit. PMID- 11040915 TI - Audit of small-incision cataract surgery using an anterior chamber maintainer. AB - PURPOSE: To assess the results of small-incision cataract surgery with intraocular lens implantation using an anterior chamber maintainer (ACM) performed between March 1997 and December 1998. METHOD: A retrospective audit was performed of all 300 consecutive patients who underwent extracapsular cataract extraction using a 6 mm scleral tunnel incision, anterior chamber maintainer and manual fragmentation of the nucleus. RESULTS: Ninety per cent of patients had a gain in visual acuity at the end of 3 months. The rate of posterior capsule opacification was comparable to the results of the National Cataract Surgery Survey (RCO 1993), i.e. 13%, but the rate of corneal endothelial decompensation and endophthalmitis was marginally higher. CONCLUSION: Appropriate selection of cases, meticulous wound closure and subconjunctival antibiotics at the end of surgery make this an acceptable alternative small-incision closed-system low-cost procedure where phacoemulsification is not available. PMID- 11040916 TI - Bilateral idiopathic retinal telangiectasis, progressive neuroradiological abnormalities and ectodermal dysplasia. PMID- 11040917 TI - Indocyanine green angiography in a case of idiopathic retinal vasculitis, aneurysms and neuroretinitis. PMID- 11040918 TI - Stable pigmentary retinopathy in a child with 3-hydroxyacyl-CoA dehydrogenase deficiency. PMID- 11040919 TI - Spontaneous reattachment of retinal detachment in a highly myopic eye with a macular hole. PMID- 11040920 TI - Central retinal vein occlusion complicating treatment with intravenous immunoglobulin. PMID- 11040921 TI - Pigment dispersion syndrome and butterfly-shaped pattern dystrophy of the retinal pigment epithelium. PMID- 11040922 TI - Null cell lymphoblastic lymphoma of the orbit. PMID- 11040923 TI - Metastatic cutaneous melanoma to the conjunctiva in an Afro-Caribbean patient. PMID- 11040924 TI - Late infection of a 5-fluorouracil enhanced bleb following systemic chemotherapy. PMID- 11040925 TI - Ophthalmic findings in HIV seropositive Tanzanian patients. PMID- 11040926 TI - Endogenous Aspergillus endophthalmitis occurring in a child with normal immune function. PMID- 11040927 TI - Bilateral reversible optic disc oedema associated with iron deficiency anaemia. PMID- 11040928 TI - Partial unilateral third nerve palsy and bilateral internuclear ophthalmoplegia: an unusual presentation of multiple sclerosis. PMID- 11040929 TI - A family with Rieger's syndrome and aniridia. PMID- 11040930 TI - Granulomatous orbital myositis. PMID- 11040931 TI - Polyhexamethylene biguanide (0.02%) alone is not adequate for treating chronic Acanthameoba keratitis. PMID- 11040932 TI - Peripheral idiopathic polypoidal choroidal vasculopathy. PMID- 11040933 TI - Idiopathic sclerochoroidal calcification. PMID- 11040934 TI - Para-lenticular metallic foreign body missed by high-resolution computed tomography. PMID- 11040935 TI - Acute acquired comitant esotropia: a prospective study. PMID- 11040936 TI - Pre-operative malposition of foldable implants. PMID- 11040938 TI - Retroviral vectors. AB - Traditionally, the retrovirus is regarded as an enemy to be overcome. However, for the past two decades retroviruses have been harnessed as vehicles for transferring genes into eukaryotic cells, a process known as transduction. During this time, the technology has moved from being a scientific laboratory tool to a potential clinical molecular medicine to be used in gene therapy. This review explains the strategy for harnessing the retrovirus life cycle, the scientific research and clinical applications of this methodology, and its limitations, as well as possible future developments. PMID- 11040937 TI - Molecular pathogenesis of oral squamous carcinoma. AB - Oral squamous carcinogenesis is a multistep process in which multiple genetic events occur that alter the normal functions of oncogenes and tumour suppressor genes. This can result in increased production of growth factors or numbers of cell surface receptors, enhanced intracellular messenger messenger signalling, and/or increased production of transcription factors. In combination with the loss of tumour suppressor activity, this leads to a cell phenotype capable of increased cell proliferation, with loss of cell cohesion, and the ability to infiltrate local tissue and spread to distant sites. Recent advances in the understanding of the molecular control of these various pathways will allow more accurate diagnosis and assessment of prognosis, and might lead the way for more novel approaches to treatment and prevention. PMID- 11040939 TI - Demystified ... microsatellites. AB - Microsatellite DNA sequences consist of relatively short repeats of one to five base pair units; together with satellites and minisatellites they comprise a larger family known as tandemly repetitive sequences. Microsatellites are found both in prokaryotes and eukaryotes, including humans, wherein they appear scattered almost at random throughout the genome. Although in prokaryotes distinct biological functions have been demonstrated, the role of microsatellites in eukaryotes is less clear. Nevertheless, several interesting hypotheses exist suggesting that certain microsatellites may exert subtle influences on the regulation of gene expression. Although the presence of these subtle mechanisms may be beneficial to a whole population, when they go wrong, as is thought to happen in the case of human trinucleotide repeat associated diseases, such as Huntington's disease, the consequences for the individual can be fatal. Most human microsatellites probably have no biological use at all; however, they are extremely useful in such fields as forensic DNA profiling and genetic linkage analysis, which can be used to search for genes involved in a wide range of disorders. With a primary focus on humans, it is the aim of this review to present an up to date discussion, both of the biological aspects and scientific uses of microsatellite sequences. In the latter case, basic theoretical and technical points will be considered, and as such it may be of use both to laboratory and non-laboratory based readers. PMID- 11040940 TI - Frequent loss of heterozygosity on chromosome 5 in non-small cell lung carcinoma. AB - AIMS: Loss of heterozygosity (LOH) at specific chromosomal regions strongly suggests the existence of tumour suppressor genes at the relevant segment. Frequent LOH on chromosome 5q has been reported in a wide variety of human tumours, including those of the lung. The aim of this study was to screen for LOH and to clarify the location of putative tumour suppressor genes on chromosome 5 implicated in the genesis and/or development of non-small cell lung carcinoma. METHODS: Thirty three patients with advanced non-small cell lung carcinoma were screened for LOH with a panel of 21 microsatellite DNA markers spanning the entire chromosome 5, using semi-automated fluorochrome based methodology. RESULTS: Twenty of the non-small cell lung carcinoma samples displayed LOH for one or more informative locus. LOH involving only 5q was found in 10 of 14 of the informative samples. Deletions involving 5p only were not present in the samples under study. There was no evidence of microsatellite instability in any of the analysed loci. These results indicate the presence of five distinct segments displaying high frequencies of deletion on chromosome 5, namely: 5q11.2-q12.2, 5q15 (D5S644 locus), 5q22.3-q23.1, 5q31.1, and 5q35.3. Eight of 14 samples had simultaneous interstitial deletions in at least two different regions. Moreover, concomitant deletion of three and four distinct regions was displayed in three of 14 and two of 14, respectively, of the informative samples. CONCLUSION: Allelic deletion on chromosome 5 is a frequent event in patients with non-small cell lung carcinoma. These results suggest the involvement of these five regions, either independently or simultaneously, in both lung squamous cell carcinoma and lung adenocarcinoma. PMID- 11040941 TI - Detection of c-kit mutation Asp 816 to Val in microdissected bone marrow infiltrates in a case of systemic mastocytosis associated with chronic myelomonocytic leukaemia. AB - BACKGROUND/AIMS: The occurrence of myeloid leukaemia in patients with systemic mastocytosis is a well recognised phenomenon. However, the pathophysiological basis of such a coevolution has not been clarified. Recent data have shown that the c-kit mutation Asp 816 to Val is detectable in neoplastic mast cells in most patients with systemic mastocytosis, including those who have associated haematological disorders. The aim of this study was to study clonal disease evolution by analysing bone marrow cells from a patient with systemic mastocytosis and associated chronic myelomonocytic leukaemia (CMML) for the presence of this mutation. METHODS: The DNA of microdissected bone marrow cells from a patient with systemic mastocytosis and associated CMML was analysed for the presence of the c-kit mutation Asp 816 to Val by means of HinfI digestion and direct sequencing of semi-nested polymerase chain reaction (PCR) products. RESULTS: The two neoplasms could easily be identified and discriminated in paraffin wax embedded bone marrow sections by tryptase and chloroacetate esterase staining. A total number of 10 tryptase positive systemic mastocytosis infiltrates and 10 tryptase negative CMML infiltrates were removed by microdissection. As assessed by HinfI digestion and direct sequencing of semi nested PCR products, the c-kit mutation Asp 816 to Val was detected in five of seven systemic mastocytosis infiltrates and four of six CMML infiltrates. By contrast, no c-kit mutation Asp 816 to Val was found in bone marrow infiltrates in patients with CMML without associated systemic mastocytosis (n = 20). CONCLUSION: These data support a monoclonal evolution of systemic mastocytosis and concurrent CMML in the patient studied. PMID- 11040942 TI - Limitations of clonality analysis of B cell proliferations using CDR3 polymerase chain reaction. AB - BACKGROUND/AIMS: Detection of clonal immunoglobulin heavy chain (IgH) rearrangements by the polymerase chain reaction (PCR) is an attractive alternative to Southern blotting in lymphoma diagnostics. However, the advantages and limitations of PCR in clonality analysis are still not fully appreciated. In this study, clonality was analysed by means of PCR, focusing in particular on the sample size requirements when studying extremely small samples of polyclonal and monoclonal lesions. MATERIALS/METHODS: High resolution complementarity determining region 3 (CDR3) PCR was used to investigate the minimum number of cells and the amount of tissue required for the detection of a polyclonal population, both for fresh cells and formalin fixed, paraffin wax embedded tissue. Subsequently, frozen and paraffin wax embedded samples of 76 B cell lymphoproliferative disorders, 43 of which were tested by means of Southern blotting, were analysed to establish the sensitivity of this assay. These specimens included 12 chronic lymphocytic leukaemias (CLLs), nine mantle cell lymphomas (MCLs), 10 follicular lymphomas (FLs), and 45 mucosa associated lymphoid tissue (MALT) lymphomas. The specificity was tested on reactive lymph nodes (n = 19), tonsils (n = 4), peripheral blood lymphocyte fractions (n = 4), and biopsies with gastritis (n = 21). RESULTS: In reactive tissue, 20 ng of high molecular weight DNA derived from 6.5-9 x 10(3) B cells was sufficient to obtain a polyclonal PCR result. With smaller amounts "pseudoclonality" could be induced. When using paraffin wax blocks, undiluted DNA isolated from tonsillar tissue of at least 1 mm2 was necessary to obtain a polyclonal pattern. The sensitivity required to detect clonality in paraffin wax embedded and frozen tissue by PCR for FL (40% and 60%, respectively) was lower than that for MALT lymphomas (60% and 86%, respectively), CLL (78% and 89%, respectively), and MCL (88% and 100%, respectively). PCR specificity was 96% and 100% for frozen and paraffin wax embedded tissue, respectively. CONCLUSION: The minimum amount of template for CDR3 PCR is approximately 20 ng of high molecular weight DNA or 1 mm3 of B cell rich paraffin wax embedded normal tonsillar tissue, but care has to be taken to avoid pseudoclonality when low numbers of B cells are present. Duplicate or triplicate tests should be performed to avoid misinterpretation. The specificity of the PCR assay is almost 100%, whereas sensitivity depends on a combination of factors, such as lymphoma type and tissue fixation. Because frozen samples yield better results, obtaining fresh material for the PCR assay is recommended, especially when analysing FL and MALT lymphomas. PMID- 11040943 TI - HPV-16 E2 gene disruption and sequence variation in CIN 3 lesions and invasive squamous cell carcinomas of the cervix: relation to numerical chromosome abnormalities. AB - AIM: To test the hypothesis that, because the human papillomavirus (HPV) E2 protein represses viral early gene transcription, E2 gene sequence variation or disruption could play a part in the induction of the numerical chromosome abnormalities that have been described in squamous cervical lesions. METHODS: The integrity and sequence of the E2 gene from 11 cervical intraepithelial neoplasia (CIN) grade 3 lesions and 14 invasive squamous cell carcinomas, all of which contained HPV-16, were analysed by the polymerase chain reaction (PCR). The E2 gene was amplified in three overlapping fragments and PCR products sequenced directly. Chromosome abnormalities were identified by interphase cytogenetics using chromosome specific probes for chromosomes 1, 3, 11, 17, 18, and X. RESULTS: E2 gene disruption was present in significantly more invasive carcinomas (eight of 14) than CIN 3 lesions (one of 11) (p = 0.03). No association was found between E2 disruption and the presence of a numerical chromosome abnormality. The E2 gene from the non-disrupted isolates was sequenced and wild-type (n = 5) and variant (n = 11) sequences identified. Variant sequences belonged to European and African classes and contained from one to 15 amino acid substitutions. Although numerical chromosome abnormalities were significantly more frequent in invasive squamous cell carcinoma than CIN 3 (p = 0.04), there was no significant relation between the presence of sequence variation and either histological diagnosis or chromosome abnormality. CONCLUSIONS: These data do not support the hypothesis that E2 gene disruption or variation is important in the induction of chromosome imbalance in these lesions. However, there is a relation between E2 gene disruption and the presence of invasive disease. PMID- 11040944 TI - The p53 gene in patients under the age of 40 with gastric cancer: mutation rates are low but are associated with a cardiac location. AB - BACKGROUND: Determining both the frequency and the spectrum of p53 gene mutation in young patients with gastric cancer might provide clues to the host related genetic mechanism(s) in gastric carcinogenesis. PATIENTS AND METHODS: p53 mutations were assessed (by means of polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), followed by DNA sequencing) in a cohort of 105 consecutive Italian patients in whom gastric cancer was ascertained before the age of 41. RESULTS: A low prevalence of p53 mutations (eight of 105) was observed, with no significant difference between intestinal (three of 31; 10%) and diffuse (five of 74; 7%) phenotypes. A significantly higher prevalence of p53 mutations was associated with the cardiac location (odds ratio, 7.09; confidence interval, 1.56 to 32.11). In all but one case, p53 mutations were associated with a stage higher than I. All eight mutations were located at CpG sites, where G : C to A : T transitions have been associated with frequent methylation at the C5 position of cytosine. CONCLUSIONS: These findings show that, unlike what has been consistently demonstrated in the general population, p53 mutations are uncommon in gastric cancer occurring in young patients, and in such patients, p53 alterations are significantly associated with the cardiac location. PMID- 11040945 TI - Identification by 16S ribosomal RNA gene sequencing of an Enterobacteriaceae species from a bone marrow transplant recipient. AB - AIMS: To ascertain the clinical relevance of a strain of Enterobacteriaceae isolated from the stool of a bone marrow transplant recipient with diarrhoea. The isolate could not be identified to the genus level by conventional phenotypic methods and required 16S ribosomal RNA (rRNA) gene sequencing for full identification. METHODS: The isolate was investigated phenotypically by standard biochemical methods using conventional biochemical tests and two commercially available systems, the Vitek (GNI+) and API (20E) systems. Genotypically, the 16S bacterial rRNA gene was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the GenBank database by multiple sequence alignment. RESULTS: Conventional biochemical tests did not reveal a pattern resembling any known member of the Enterobacteriaceae family. The isolate was identified as Salmonella arizonae (73%) and Escherichia coli (76%) by the Vitek (GNI+) and API (20E) systems, respectively. 16S rRNA sequencing showed that there was only one base difference between the isolate and E coli K-12, but 48 and 47 base differences between the isolate and S typhimurium (NCTC 8391) and S typhi (St111), respectively, showing that it was an E coli strain. The patient did not require any specific treatment and the diarrhoea subsided spontaneously. CONCLUSIONS: 16S rRNA gene sequencing was useful in ascertaining the clinical relevance of the strain of Enterobacteriaceae isolated from the stool of the bone marrow transplant recipient with diarrhoea. PMID- 11040947 TI - A memo on Meno. PMID- 11040946 TI - Screening of the entire coding region of p53 in low grade lymphoproliferative disorders. AB - This report details a rapid method for screening the entire p53 coding region (exons 2-11). This method, based on the non-isotopic RNase cleavage assay, uses novel primer sequences and an adaptation of the MutationScreener method. A mutation in 20% of the sample was easily detectable by this method, whereas mutations below 50% were undetectable using the original method. Alterations to the wild-type p53 mRNA sequence were found in nine of the 130 patients with low grade lymphoproliferative disorders screened, and this was confirmed by DNA sequencing in eight of eight samples. The method is a simple and reliable technique for screening for p53 mutations. PMID- 11040948 TI - Images, imagination, and movement: pictorial representations and their development in the work of James Gibson. AB - For more than 30 years James Gibson studied pictures and he studied motion, particularly the relationship between movement through an environment and its visual consequences. For the latter, he also struggled with how best to present his ideas to students and fellow researchers, and employed various representations and formats. This article explores the relationships between the concepts of the fidelity of pictures (an idea he first promoted and later eschewed) and evocativeness as applied to his images. Gibson ended his struggle with an image of a bird flying over a plane surrounded by a spherical representation of a vector field, an image high in evocativeness but less than completely faithful to optical flow. PMID- 11040949 TI - Depth discrimination from shading under diffuse lighting. AB - The human visual system has a remarkable ability to interpret smooth patterns of light on a surface in terms of 3-D surface geometry. Classical studies of shape from-shading perception have assumed that surface irradiance varies with the angle between the local surface normal and a collimated light source. This model holds, for example, on a sunny day. One common situation in which this model fails to hold, however, is under diffuse lighting such as on a cloudy day. Here we report on the first psychophysical experiments that address shape-from-shading under a uniform diffuse-lighting condition. Our hypothesis was that shape perception can be explained with a perceptual model that "dark means deep". We tested this hypothesis by comparing performance in a depth-discrimination task to performance in a brightness-discrimination task, using identical stimuli. We found a significant correlation between responses in the two tasks, supporting a dark-means-deep model. However, overall performance in the depth-discrimination task was superior to that predicted by a dark-means-deep model. This implies that humans use a more accurate model than dark-means-deep to perceive shape-from shading under diffuse lighting. PMID- 11040950 TI - Mechanisms for seeing transparency-from-motion and orientation-from-motion. AB - Structure-from-motion (SFM) perception is hypothesised to be mediated by units that sense the near-far relationships in transparency-from-motion (TFM) and orientation-from-motion (OFM). The frequency of subjective reversals during observation of ambiguous SFM displays is considerably decreased when either the direction of rotation or the surface orientation is oscillated during the inspection. Such manipulations impede adaptation of units selectively sensitive to TFM and OFM. The results show that both OFM and TFM units are direction selective, and that the reversal rate is unaffected by reducing TFM to zero; and support the view that depth order, when both TFM and OFM are present, is estimated by common neural units. PMID- 11040951 TI - The role of attention in motion extrapolation: are moving objects 'corrected' or flashed objects attentionally delayed? AB - Objects flashed in alignment with moving objects appear to lag behind [Nijhawan, 1994 Nature (London) 370 256-257]. Could this 'flash-lag' effect be due to attentional delays in bringing flashed items to perceptual awareness [Titchener, 1908/1973 Lectures on the Elementary Psychology of Feeling and Attention first published 1908 (New York: Macmillan); reprinted 1973 (New York: Arno Press)]? We overtly manipulated attentional allocation in three experiments to address the following questions: Is the flash-lag effect affected when attention is (a) focused on a single event in the presence of multiple events, (b) distributed over multiple events, and (c) diverted from the flashed object? To address the first two questions, five rings, moving along a circular path, were presented while observers attentively tracked one or multiple rings under four conditions: the ring in which the disk was flashed was (i) known or (ii) unknown (randomly selected from the set of five); location of the flashed disk was (i) known or (ii) unknown (randomly selected from ten locations). The third question was investigated by using two moving objects in a cost-benefit cueing paradigm. An arrow cued, with 70% or 80% validity, the position of the flashed object. Observers performed two tasks: (a) reacted as quickly as possible to flash onset; (b) reported the flash-lag effect. We obtained a significant and unaltered flash lag effect under all the attentional conditions we employed. Furthermore, though reaction times were significantly shorter for validly cued flashes, the flash-lag effect remained uninfluenced by cue validity, indicating that quicker responses to validly cued locations may be due to the shortening of post-perceptual delays in motor responses rather than the perceptual facilitation. We conclude that the computations that give rise to the flash-lag effect are independent of attentional deployment. PMID- 11040952 TI - Can semantic knowledge influence motion correspondence? AB - Semantic factors are presumed to have little influence on motion perception. Two experiments examined the effects of an object's semantic identity on motion correspondence using the Ternus paradigm. Motion correspondence was not influenced by whether the object depicted is typically moving or stationary, but it was influenced by the way(s) in which an object's components typically move relative to one another: perceived correspondence differed depending on whether the motion tokens constituted the feet of a person walking or the wheels of a car. Apparently, semantic knowledge can influence motion correspondence, although such influence is weak and may be restricted to certain types of semantic information. The adaptive significance of such restricted influences is considered. PMID- 11040953 TI - Spatial factors of brightness illusion in the Ehrenstein figure. AB - The strength of brightness illusion in an Ehrenstein figure has been examined as a function of two variables--inducing line length and gap size--by a two alternative forced-choice procedure. The results show an interaction between the two variables; the length of the inducers and gap size are both involved in the formation of the brightness illusion, with gap size as the stronger factor. A spatial limit (corresponding to a gap size of 2.4 deg) was found, below which the illusion is always present regardless of the length of the inducers. An area ratio, defined as the ratio of the area of the ring formed by the four inducing lines of an Ehrenstein figure to the area of the illusory surface, takes into account the different spatial factors studied in the experiment and the global size of the Ehrenstein figure. The strength of the illusion was found to increase linearly with increasing values of the area ratio. PMID- 11040954 TI - Integration biases in the Ouchi and other visual illusions. AB - A texture pattern devised by the Japanese artist H Ouchi has attracted wide attention because of the striking appearance of relative motion it evokes. The illusion has been the subject of several recent empirical studies. A new account is presented, along with a simple experimental test, that attributes the illusion to a bias in the way that local motion signals generated at different locations on each element are combined to code element motion. The account is generalised to two spatial illusions, the Judd illusion and the Zollner illusion (previously considered unrelated to the Ouchi illusion). The notion of integration bias is consistent with recent Bayesian approaches to visual coding, according to which the weight attached to each signal reflects its reliability and likelihood. PMID- 11040955 TI - Does face recognition rely on encoding of 3-D surface? Examining the role of shape-from-shading and shape-from-stereo. AB - It is now well known that processing of shading information in face recognition is susceptible to bottom lighting and contrast reversal, an effect that may be due to a disruption of 3-D shape processing. The question then is whether the disruption can be rectified by other sources of 3-D information, such as shape from-stereo. We examined this issue by comparing identification performance either with or without stereo information using top-lit and bottom-lit face stimuli in both photographic positive and negative conditions. The results show that none of the shading effects was reduced by the presence of stereo information. This finding supports the notion that shape-from-shading overrides shape-from-stereo in face perception. Although shape-from-stereo did produce some signs of facilitation for face identification, this effect was negligible. Together, our results support the view that 3-D shape processing plays only a minor role in face recognition. Our data are best accounted for by a weighted function of 2-D processing of shading pattern and 3-D processing of shapes, with a much greater weight assigned to 2-D pattern processing. PMID- 11040956 TI - Cross-modal interaction between vision and touch: the role of synesthetic correspondence. AB - At each moment, we experience a melange of information arriving at several senses, and often we focus on inputs from one modality and 'reject' inputs from another. Does input from a rejected sensory modality modulate one's ability to make decisions about information from a selected one? When the modalities are vision and hearing, the answer is "yes", suggesting that vision and hearing interact. In the present study, we asked whether similar interactions characterize vision and touch. As with vision and hearing, results obtained in a selective attention task show cross-modal interactions between vision and touch that depend on the synesthetic relationship between the stimulus combinations. These results imply that similar mechanisms may govern cross-modal interactions across sensory modalities. PMID- 11040957 TI - The biochemistry of aging. AB - Although philosophers and scientists have long been interested in the aging process, general interest in this fascinating and highly important topic was minimal before the 1960s. In recent decades, however, interest in aging has greatly accelerated, not only since the elderly form an ever-increasing percentage of the population, but because they utilize a significant proportion of the national expenditures. In addition, many people have come to the realization that one can now lead a very happy, active, and productive life well beyond the usual retirement age. Scientifically, aging is an extremely complex, multifactorial process, and numerous aging theories have been proposed; the most important of these are probably the genomic and free radical theories. Although it is abundantly clear that our genes influence aging and longevity, exactly how this takes place on a chemical level is only partially understood. For example, what kinds of genes are these, and what proteins do they control? Certainly they include, among others, those that regulate the processes of somatic maintenance and repair, such as the stress-response systems. The accelerated aging syndromes (i.e., Hutchinson-Gilford, Werner's, and Down's syndromes) are genetically controlled, and studies of them have decidedly increased our understanding of aging. In addition, C. elegans and D. melanogaster are important systems for studying aging. This is especially true for the former, in which the age-1 mutant has been shown to greatly increase the life span over the wild-type strain. This genetic mutation results in increased activities of the antioxidative enzymes, Cu Zn superoxide dismutase and catalase. Thus, the genomic and free radical theories are closely linked. In addition, trisomy 21 (Down's syndrome) is characterized by a significantly shortened life span; it is also plagued by increased oxidative stress which results in various free radical-related disturbances. Exactly how this extra chromosome results in an increased production of reactive oxygen species is, however, only partially understood. There is considerable additional indirect evidence supporting the free radical theory of aging. Not only are several major age-associated diseases clearly affected by increased oxidative stress (atherosclerosis, cancer, etc.), but the fact that there are numerous natural protective mechanisms to prevent oxyradical-induced cellular damage speaks loudly that this theory has a key role in aging [the presence of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, among others; various important intrinsic (uric acid, bilirubin, -SH proteins, glutathione, etc.) and extrinsic (vitamins C, E, carotenoids, flavonoids, etc.) antioxidants; and metal chelating proteins to prevent Fenton and Haber-Weiss chemistry]. In addition, a major part of the free radical theory involves the damaging role of reactive oxygen species and various toxins on mitochondria. These lead to numerous mitochondrial DNA mutations which result in a progressive reduction in energy output, significantly below that needed in body tissues. This can result in various signs of aging, such as loss of memory, hearing, vision, and stamina. Oxidative stress also inactivates critical enzymes and other proteins. In addition to these factors, caloric restriction is the only known method that increases the life span of rodents; studies currently underway suggest that this also applies to primates, and presumably to humans. Certainly, oxidative stress plays an important role here, although other, as yet unknown, factors are also presumably involved. Exactly how the other major theories (i.e., immune, neuroendocrine, somatic mutation, error catastrophe) control aging is more difficult to define. The immune and neuroendocrine systems clearly deteriorate with age. (ABSTRACT TRUNCATED) PMID- 11040958 TI - Cystatin C--properties and use as diagnostic marker. PMID- 11040959 TI - The role of receptors in prostate cancer. PMID- 11040960 TI - Oxidative modifications of protein structures. PMID- 11040961 TI - Selected markers of bone biochemistry. PMID- 11040962 TI - Laboratory markers of ovarian function. PMID- 11040963 TI - [Conformational features of a pentapeptide as an element of the active center of human immunoglobulin E]. AB - The spatial structures of human immunoglobulin E pentapeptide Asp-Ser-Asp-Pro-Arg and some of its related peptides were investigated by the method of theoretical conformational analysis. These synthetic peptides have the capacity to inhibit the binding of immunoglobulin E to the mast cells of the skin. The results of the calculations and the data on biological activity of these peptides were used for determination their energy-dependent conformational characteristics that provide their specific interaction with receptors of mast cells. PMID- 11040964 TI - [Study of the complex formation of daunomycin with deoxytetranucleotides with bases of differing sequence in an aqueous solution by 1H-NMR spectroscopy]. AB - The complex formation of the antibiotic daunomycin with deoxytetranucleotides of different base sequence in the chain, 5'-d(GpCpGpC), 5'-d(CpGpCpG), and 5' d(TpGpCpA) in aqueous salt solution was studied by 1D and 2D (2M-TOCSY and 2M NOESY) 1H-NMR spectroscopy. Concentration and temperature dependences of proton chemical shifts of molecules were measured. Based on these dependences, reaction equilibrium constants, relative content of various complexes depending on concentration and temperature, limiting values of chemical shifts of protons of daunomycin incorporated in various complexes, and the thermodynamic parameters delta H and delta S of complex formation were calculated. The analysis of the results enables the conclusion that the sites of predominant intercalation of daunomycin are triplet nucleotide sequences, the binding sites of the antibiotic with three consecutive GC pairs in the tetranucleotide duplex being more preferential. Daunomycin exhibits no sequence specificity upon binding to the single-stranded deoxynucleotide sequence. From the calculated values of induced chemical shifts of daunomycin protons and 2M-NOE data, the most probable spatial structures of complexes (1:2) of the antibiotic with deoxytetranucleotides were constructed. The binding of the second daunomycin molecule to both the single stranded and duplex form of tetramers is of pronounced anticooperative mode, which is explained by the presence in the antibiotic of a positively charged amino sugar residue, which poses considerable steric constraints for the insertion of the second antibiotic molecule into the short tetranucleotide sequence. The results were compared with the data obtained under identical experimental conditions for typical intercalators proflavine and ethidium bromide. PMID- 11040965 TI - [Interaction of Ag+ ions with DNA and its monomers]. AB - Differential UV spectra of DNA and its monomers that were induced by Ag+ ions were measured, and the effect of ions on the parameters of the helix-coil transition was studied. The data obtained confirm the existence of "strong" and "weak" modes of binding of Ag+ to DNA. The earlier proposed proton transfer from N1G to N3C, which is determined by the interaction of Ag+ with N7G (a "strong" complex), follows immediately from the shape of the differential UV spectra. The positive cooperativity of the binding of Ag+ to DNA upon the formation of a "weak" complex is due to the cooperativity of the transition of DNA to a new double-helical conformation. A model of this conformation is proposed which suggests the formation of Hougsteen GC and AT pairs. PMID- 11040966 TI - [Interaction of eukaryotic DNA with apolipoprotein A-I and its complexes with glucocorticoids]. AB - A biochemically active complex of apolipoprotein A-I with tetrahydrocortisol was revealed, which increases gene expression in hepatocytes. It was shown by the method of fluorescent probe that titration of rat liver DNA by the apolipoprotein A-I-tetrahydrocortisol complex leads to the appearance of single-stranded fragments. The effect of the complex on the secondary structure of native DNA was confirmed by the method of small-angle X-ray scattering. It was shown that approximately 54 apolipoprotein A-I molecules carrying tetrahydrocortisol as a ligand bind to one molecule of isolated native DNA, inducing a break of hydrogen bonds between the pair of nitrous bases. It is concluded that the cooperative effect of high-density lipoproteins and cortisol in the regulation of gene expression in hepatocytes with the participation of resident liver macrophages is accomplished by a new biochemical mechanism. This mechanism makes itself evident as a result of the interaction of DNA with the apolipoprotein A-I tetrahydrocortisol complex, the appearance of single-stranded DNA regions in binding sites, and subsequent initiation of gene transcription. PMID- 11040967 TI - [Nonlinear elasticity and dynamics of globular proteins]. AB - A molecule of native protein exhibits a high degree of mechanical nonlinearity, which gives rise to a peculiar dynamics behavior of globular proteins. Intramolecular motions strongly depend on pressure. The reaction of specific volume to the action of electric field (electrostriction) inside the molecule has a sign opposite to that observed in liquids. The probability that a small ion would penetrate inside the globule from the solvent depends nonmonotonously on hydrostatic pressure. The nonmonotonicity should manifest itself in all processes related to the transfer of ions inside the globule. The relationship between the mechanical properties of the protein and the kinetics of hydrogen exchange at normal and high pressure is discussed. PMID- 11040968 TI - [Gravitational cause of fluctuations of the rate of oxidation of unithiol by nitrite ion]. AB - The rate of oxidation of unithiol (sodium dimercaptopropansulfonate) by nitrite ion was determined in the course of the annual experiment at the Antarctic station Mirny in 1996-1997. The rhythmic fluctuations in the oxidation rate were found. It is shown that these fluctuations correlate with changes in the velocity of the Earth's forward-rotational movement by the action of the Sun and Moon. PMID- 11040969 TI - [Study of the dielectric permeability in the superhigh frequency range of a degraded polyoxybutyrate biopolymer]. AB - The dielectric permeability of the degradable biopolymer polyhydroxybutyrate synthesized by hydrogen-oxidizing bacteria Alcaligenes eutrophus was investigated by the resonance method using original highly sensitive microstrip sensors. For the first time, a linear growth of dielectric permeability (delta epsilon/delta T = 7 x 10(-4) degree-1) due to the flexibility of the polymer chain in the temperature range from 10 to 70 degrees C was revealed. The energy of a bend of the nearest fragments was evaluated (E = 392 K), and its correspondence to the energies of bends of the alcyl groups of low-molecular substances like liquid crystals was established. It was shown that at low values of dielectric permeability in the high-frequency range (epsilon' = 2.25 +/- 0.02), which are stable, in a wide range of frequencies of the electromagnetic field (1 MHz - 1 Hz), polyoxybutyrate can be used in the microwave equipment. PMID- 11040970 TI - [Comparative study of the physico-chemical properties of chitosans with varying degree of polymerization in neutral aqueous solutions]. AB - The physicochemical properties of chitosan samples with high (130 kD) and low (30 kD) molecular masses in neutral aqueous solutions (pH 6.0) were studied by the methods of high-speed and equilibrium sedimentation, viscosimetry, and NMR and UV spectroscopies. Differences in the hydrodynamic characteristics of the samples were revealed. It was found that low-molecular-weight chitosan represents flexible linear macromolecules which undergo conformational changes upon temperature increase. The high-molecular-weight chitosan forms more rigid asymmetric structures whose conformation does not vary significantly with temperature increase. It was found that the high-molecular-weight chitosan has a higher constant of binding to the anionic dye tropeoline 000-II, which can be explained by different conformations of their macromolecules in solution. PMID- 11040971 TI - [Effect of dipyridamole on recombination of photooxidized bacteriochlorophylla and photoreduced primary quinone in reactive centers of purple bacteria and degradation of form M412 of bacteriorhodopsin]. AB - It is shown that the addition of dipyridamole (2,6-bis(diethanolamino)-4,8 dipiperidinopyrimido[5,4d]py rim idine) (up to 10(-4) M) leads to a drastic acceleration of the dark recombination reaction between photooxidized bacteriochlorophyll and photoreduced primary quinone in reaction centers of Rhodobacter sphaeroides. The value of the acceleration is similar to that registered under cryogenic temperatures. The extent of the effect of dipyridamole derivatives depended on their structure. In wild-type bacteriorhodopsin and D96N mutant, dipyridamole slowed down the Schiff base reprotonation (the kinetics of M412 form decay was registered). It is suggested that dipyridamole can influence the structural and dynamic state of membrane proteins by affecting the system of their hydrogen-bonds and thus modify electron and proton transport processes. PMID- 11040972 TI - [Effect of lipid monolayers on diffusion of oxygen through the air/water interface]. AB - The dependence of the diffusion current on the depth of immersion of the electrode was studied by polarography using an open platinum electrode. As the electrode was brought from the depth of the liquid phase to its surface, an increase in the current under aerobic conditions was observed, due to diffusion of oxygen through the interface. The formation of lipid monolayers of phosphatidylcholine, stearic acid, hexadecanol, octadecanol, eicosanol, and docosanol on the water surface led to a decrease in diffusion current; the effect being most pronounced at a minimal depth of immersion of the electrode. The maximum value of the relative decrease in diffusion current R was obtained for docosanol monolaers. It was shown that the R value increases with increasing surface pressure in monolayers of phosphatidylcholine and stearic acid. It is assumed that the decrease in diffusion flow of O2 in the presence of monolayers is caused by the formation of an energy barrier that prevents the sorption of O2, which is related to the presence of hydrocarbon chains weakly interacting with oxygen. PMID- 11040973 TI - [Comparative effectiveness of antioxidants in protecting the bacterial plasmatic membrane from the active froms of oxygen]. AB - The effect of hydroxyl radicals OH. generated by the decomposition of H2O2 by Fe2+ ions (Fenton reaction) on the barrier properties of plasma membranes of Escherichia coli cells K-12 was studied by electroorientation spectroscopy. It was found that the administration of hydrogen peroxide led to the disturbance of the barrier properties of plasma membranes only when the cells were preincubated with Fe2+ ions and their constant concentration in the system was maintained by ascorbate or dithiotreitol (150-500 microM). The extent of the toxic action on plasma membranes depended on the concentration of reacting elements and the substance used as a reducer Fe2+. The efficiency of protection of antioxidants of different classes (enzymic, SH-containing, and phenolic compounds) against the toxic action of hydroxyl radicals on plasmatic membranes was shown. PMID- 11040974 TI - [Effect of products of radiation-induced free radical fragmentation of phospholipids and temperature on lipid membranes]. AB - Thermotropic behavior of liposomes exposed to gamma-radiation was studied by differential scanning microcalorimetry. It was found that the peak corresponding to the gel-liquid crystal transition for liposomes composed of bovine brain sphingomyelin and dipalmitoyl phosphatidylglycerol broadened and shifted toward the high-temperature region. No effect of irradiation on dipalmitoyl phosphatidylcholine liposomes was observed. Previously it was shown that, on exposure to gamma-rays, sphingomyelin and phosphatidylglycerol, as opposed to phosphatidylcholine, broke down into fragments of lower molecular weight. It is concluded that the accumulation of products of phospholipid fragmentation in the membrane results in the changes of phase transition parameters. PMID- 11040975 TI - [Effect of ascorbic acid on production of nitric oxide by leukocytes]. AB - The effect of ascorbic acid on suspensions of blood formic elements was studied by the ESR method. It was shown that incubation of a suspension of formic blood elements in the presence of ascorbic acid leads to the appearance of nitric oxide, which is produced by leukocytes and partially probably by thrombocytes. The formation of nitric oxide is evidenced by the appearance of nitrosyl complexes heme-NO. In this case, hemoglobin of erythrocytes serves as a natural trap for nitric oxide. PMID- 11040976 TI - [Nonspecific potential-dependence of conductivity of a membrane, modified by nonselective channel formers with an unlimited number of states]. AB - A formula describing the nonspecific potential-dependent conductance of membranes modified by a channel former with an unlimited number of states was derived. The characteristic features of this potential dependence and their experimental manifestations were analyzed. PMID- 11040977 TI - [Dynamics of neuroreceptor function in a chemical synapse]. AB - A model for the dynamics of functioning of neuroreceptors in chemical synapses is proposed. It is shown that the functioning of a neuroreceptor by the action of mediator is determined by correlation of the mechanical elasticity of the receptor with its electrical polarizability and the presence of spiralized fragments. It is shown in terms of the quantum-mechanical approach that, as the charged groups of the mediator come close to the C-end of the neuroreceptor, a shift of potential surface occurs, which corresponds to the deformation of the receptor. The microscopic approach enables one to describe more exactly the dynamics of functioning of neuroreceptors than the macroscopic approach; however, its implementation requires the use of powerful computers. If only qualitative assessment is needed, the macroscopic approach would suffice. It is assumed that in terms of the model, the saltatory (jump-like) conduction of the spike along the axon can be explained. PMID- 11040978 TI - [Viscumin modulates neutrophil respiratory burst, caused by a chemotactic peptide]. AB - The ability of viscum at different concentrations to modulate the respiratory burst in neutrophils, induced by the chemotactic peptide N-formyl-methionyl leucyl-phenylalanine was studied. This does not exclude the possibility that viscum can interact with the receptor of this peptide. The analysis of the primary structure of viscum revealed elements structurally analogous to the chemotactic peptide. It is assumed that viscum can exhibit the properties an antagonist of the receptor of N-formyl-methionyl-leucyl-phenylalanine, and the mechanism of action of viscum depends on its concentration. PMID- 11040979 TI - [Generation of superoxide radicals by heart mitochondria: study by spin trapping under continuous oxygenation]. AB - The generation of superoxide radicals by isolated rat heart mitochondria was studied by the spin trapping technique. The sample was placed into the cavity of an EPR spectrometer in a thin-wall teflon capillary tube, which made it possible to maintain the partial oxygen pressure in the mitochondrial suspension at a constant level. Tiron was used as a spin trap, and the intensity of its EPR signal corresponded to the rate of O2-. formation in the sample. The addition of oxidation substrates (succinate, glutamate, and malate) into the incubation mixture caused the appearance of the Tiron EPR signal. The rate of superoxide radical generation by heart mitochondria strongly increased in the presence of antimycin A, an inhibitor of the Q-cycle in complex III of the respiratory chain, but it was completely depressed by another inhibitor of Q-cycle myxothiazol. The inhibition of the reverse electron transport in complex I of the respiratory chain by rotenone (oxidation substrate--succinate) caused a substantial decrease in the rate of O2-. formation by mitochondria. PMID- 11040980 TI - [Asymptotic solutions of population dynamic equations]. AB - An approach is offered to construct the asymptomatics of the solutions on the small parameter in the close neighborhood of the equilibrium condition of the well-known Volterra-Lotka "prey-predator" system and one of its modifications which takes into account the intraspecies competition of preys and limitation of food resources of a predator. On the basis of the formulas obtained, possible dynamic modes of the size of populations of both kinds are analyzed. PMID- 11040981 TI - [Fluorescence, excited by light in the 380-540 nm wavelength range, in in cucumber leaves depends on the time of vegetation and light regime]. AB - The slow fluorescence induction produced in cucumber leaves by light in the range of wavelengths 380-540 nm and intensity of 180 W/m2 was studied. The ratio of fluorescence maxima in the red region (F734/F682) in young and mature leaves was approximately 2. It is assumed that this value depends on an increase in the contribution of the long-wavelength fluorescence due to the spillover effect. In plants grown under natural conditions, the parameter F734/F682 correlated with the concentrations of chlorophyll and carotenoids. In plants grown in the light of red and blue regions no such correlation was observed and the F734/F682 remained unchanged. It is concluded that the F534/F682 is affected by the intensity and spectral composition of exciting light used during the growing. PMID- 11040982 TI - [Release of iron ions from transferrin under the effect of nitric oxide]. AB - The dynamics of EPR signals from the iron-transporting blood protein Fe(3+) transferrine after the administration of sodium nitrite and metronidazole to animals was studied. It was shown that exogenin nitric oxide produced by nitrocompounds resulted in the release of iron from Fe(3+)-transferrine. PMID- 11040983 TI - [Study of the functional status of a synthetic apparatus of blood lymphocytes under the action of weak low-frequency magnetic fields]. AB - The functional state of the synthetic apparatus of animal blood lymphocytes under weak low-frequency magnetic fields was studied. The changes in blood cell synthetic activity and leukocyte formula, a signal index of general nonspecific adaptation reactions, were shown. In the majority of cases examined, the variable magnetic fields increase the functional activity of the lymphocyte synthetic apparatus, improve the type of adaptation reaction, and enhance the level of immune resistance of the organism. PMID- 11040984 TI - [Reconstruction of elastic properties of soft biological tissues from data about it's deformation under dynamic load]. AB - The present work continues theoretical investigations on developing new noninvasive methods for diagnostics of tissue pathologies. These methods are based on tissue elasticity reconstruction using the measured internal displacements occurring due to the deformation of the object under study. Dynamic loading of the object is considered. The robustness of the proposed elasticity reconstruction procedure is demonstrated using numerical modeling of the displacement fields inside the nonhomogeneous object. PMID- 11040985 TI - [Change in the reaction of the antioxidant system of wheat sprouts after UV irradiation of seeds]. AB - The response of the antioxidant system of sprouts of wheat Triticum aestivum L. to preliminary irradiation of seeds with UV light was studied. The dependence of lipid peroxidation and the extent of antioxidant activity on the duration of irradiation was studied. It was shown that low doses of UV radiation (5-15 min) stimulate the antioxidant protection of green wheat sprouts grown for eight days. Increasing the irradiation time to 30-60 min leads to the inhibition of lipid peroxidation by the antioxidant system. A more prolonged irradiation of seeds with UV light (for 1-6 h) led to an increase in the level of lipid peroxidation in sprouts. However, 1-2-day-old sprouts from seeds irradiated for 5-6 h, adapted themselves to the influence due to the compensatory mechanisms. By the 8th day of germination of preliminarily irradiated seeds, the content of antioxidants and malone dialdehyde returned to the norm. The dynamics of activity of peroxidase in seeds irradiated with low doses of UV light for 30 min was studied. It was found that on the third day of seed germination, a decrease in peroxidase activity followed by its slight increase occurred. The maximum activity of the enzyme in the endosperm was observed on day 5-6, and in roots and green sprouts, on day 3-5 of germination. It was concluded that antioxidants and peroxidase are involved in the compensatory mechanisms of inhibition of free radicals formed upon UV irradiation of seeds. PMID- 11040986 TI - [Effect of a low-frequencey magnetic field on esterase activity and change in pH in wheat germ during swelling of wehat seeds]. AB - The role of nonsteady phenomena determined by a low velocity of ion movements in a weak external field is considered in relation to their possible nonlinear effects on processes occurring in boundary layers near the membrane, particularly, on the release of membrane-bound proteins and pH value. It is shown that a short-term treatment of wheat seeds with low-frequency magnetic field at the stage of esterase activation during seed swelling enhances the activation of esterases; the effect observed at final stages of activation depends on the time after the treatment with electromagnetic field. Treatment of seeds with electromagnetic field at this stage changed qualitatively the time course of the release of reaction products into the medium: the reaction rate increased initially and then decreased below the control level. At earlier stages of swelling in treated seeds and at all stages in control seeds, the time course of the product release was linear. The retardation of the release of the reaction products at terminal stages of esterase activation is presumably related to the release of proteins and their complexes under the action of electromagnetic field and the resulting restoration of the barrier properties of membranes. Treatment with electromagnetic field also caused a noticeable acceleration of proton flow form the medium, which was judged from pH changes in the bulk medium and in the vicinity of germ surface. The difference between the treated and control samples after 23-24 h of imbibition became statistically significant and was as high as 0.4 pH units. By taking into account the nonsteady phenomena occurring upon action of low-frequency electromagnetic field, it is possible to explain unusual dependences of biological effects on the amplitude of the electromagnetic field, including the atypical enhancement of these effects by the action of weak low frequency fields. PMID- 11040987 TI - [The role of carnivorous zooplankton in dynamics of trophic interaction of fish and plankton]. AB - A conceptual (minimal) model of the aquatic community is proposed, which includes phytoplankton, zooplankton, fish and fish larvae. It is shown that carnivorous zooplankton increases the system stability when the fish predation changes. As a result, the system collapse followed by algal bloom becomes less probable. PMID- 11040988 TI - [Traveling waves in cross-diffusion models of population dynamics]. AB - "Traveling wave"--type solutions of some models with cross-diffusion were considered. It was shown that "cross-diffusion" terms, as opposed to "diffusion" terms, do not increase the dimensionality of the automodel system. The bifurcation approach was used to study the dependence of the wave solutions on the parameters of the models being considered, and the waves were classified. It was shown that, at the same parameter values, both "fast" and "slow" waves can exist and that these waves are described by different automodel systems. PMID- 11040989 TI - [Rotation of biological systems in a magnetic field: slitting of spectra for some magnetobiological effects]. AB - The interference mechanism for biological reception of weak magnetic fields was studied with consideration for the own molecular rotations of ion-protein complexes. An additional rotation of a biological system is shown to decrease the biological effect of "magnetic vacuum" and split spectral peaks from the effects of static magnetic field. PMID- 11040990 TI - [Comparative experimental and theoretical study of the dependence of parameters of curved induction of fluorescence and millisecond delayed luminescence on dark adaptation time]. AB - The dependences of G = (Fm - Fs)/Fm parameter for slow fluorescence induction and delayed luminescence induction curves on adaptation time t(ad) were obtained both in experiment, for the leaves of Hibiscus rosa chinensis in vivo, and in a theoretical study with the help of the theoretical model developed earlier. In both cases, the G(t(ad)) dependences have the form of curves with saturation, which are well described by G = A(1 - exp(-(t(ad)/T0)). Both in experiment and in theoretical calculations, G values for delayed luminescence were larger than for fluorescence. The ratio of saturation times for G(t(ad)) for fluorescence and delayed luminescence was 1.40 for experiment and 1.46 for theoretical calculations. These results suggest that delayed luminescence induction is more sensitive to changes in the state of plant than the slow fluorescence induction. PMID- 11040991 TI - [Induction of delayed luminescence of pea photosystem II with genetically changed level of starch in the seeds]. AB - We studied the curves of delayed luminescence induction in leaves of pea plants with mutations affecting the starch branching enzyme (r locus) and ADPG pyrophosphorylase (rb locus). In mutated pants, a 75% reduction of starch content in seeds was observed. The half-decay time of delayed luminescence intensity during induction for double mutants was shown to be longer than for wild type plants. PMID- 11040992 TI - [Biological role of a neurotrophic factor fragment from pigment epithelium: structure-functional homology with a differentiation factor for the HL-60 cell line]. AB - It was shown that the full-size neurotrophic factor from pigment epithelium (PEDF) induces the cell differentiation of the human promyelocyte leukemia cell line HL-60. A structural analysis of PEDF revealed in its C-terminal region a six membered peptide fragment PEDF-(352-357) (PEDF-6) whose sequence is highly homologous to the 41-46 fragment of the active site of the human leukocyte differentiation factor HLDF (HLDF-6). The biological effect of PEDF and synthetic peptides PEDF-6 and HLDF-6 on the HL-60 cells and the early gastrula ectoderm of Xenopus laevis embryos was studied. On the basis of the structural and functional homologies of HLDF, PEDF, and their homologous peptides and the computer models of the spatial structures of the full-size PEDF and the PEDF with the C-terminal fragment split off tby the cleavage of the Leu380-Thr381 bond in the serpin loop, a hypothesis on the functional role of the serpin loop in PEDF was put forward. PMID- 11040993 TI - [Spatial structure of a Fab-fragment of a monoclonal antibody to human interleukin-2 in two crystalline forms at a resolution of 2.2 and 2.9 angstroms]. AB - The three-dimensional structure of the antigen-binding fragment of a monoclonal antibody to human interleukin-2 was determined in two crystal forms by the X-ray method of molecular replacement at 2.2 and 2.9 A resolutions. The spatial structure of the protein and the stereochemistry of its antigen-binding site were analyzed. PMID- 11040994 TI - [Identification of a fragment of ceruloplasmin, interacting with copper transporting Menkes ATPase]. AB - The interaction was studied of ceruloplasmin (Cp, EC 1.16.3.1), a copper containing plasma protein, with two synthetic peptides P15 and P16 whose structures correlate with those of the noncytosolic regions of the copper transfer P1 type ATPase (ATP7A), apparently encoded by the Menkes disease gene (Atp7a). Pentadecapeptide P15 and hexadecapeptide P16 were synthesized using the solid phase method. They correspond to fragments of two extracellular loops ATP7A, of which one loop is apparently involved in the copper ion transfer (P16) whereas the other is not (P15). The protein footprinting showed that P16 binds to a fragment of the ceruloplasmin domain 6. Kinetics of the ceruloplasmin-P16 binding was studied by affinity chromatography on P16 immobilized on a macroporous disk, and the Kd value (1.5 x 10(-6) M) of this interaction was determined. The ATP7A involvement in the copper ion transfer to nonhepatocyte cells is discussed. PMID- 11040995 TI - [Study of solid-phase catalytic isotopic exchange of hydrogen in alpha-conotoxin G1 under the effect of spillover-tritium]. AB - Tritium-labeled alpha-conotoxin G1 with a molar radioactivity of 35 Ci/mmol and full biological activity (according to the binding to nicotinic acetylcholine receptor) was obtained by the high-temperature solid-state catalytic isotope exchange (HSCIE). The tritium distribution in the molecule of alpha-conotoxin G1 was revealed by 3H NMR spectroscopy. Tritium was found in all amino acid residues except for the Asn4-Pro5-Ala6 fragment. The data on the comparative reactivity of C-H bonds, the ab initio quantum-chemical calculation of the hydrogen exchange reaction, and the information on the spatial structures of alpha-conotoxin G1 in solution and in crystal state allowed us to establish that the reactivity of H atoms may be increased by their interaction with the electron donor O and N atoms at the transition state of the HSCIE reaction. A decrease in the rate of the HSCIE reaction could be caused by both a poor spatial accessibility of C-H bonds and a limited mobility of the peptide fragment containing these bonds. PMID- 11040996 TI - [Characteristics of recombinant fragments of the protective antigen SPA of epidemic typhus pathogens]. AB - Fragments of a gene for species-specific protective antigen SPA of Rickettsia prowazekii earlier cloned in lambda gt11 were recloned into the in-frame expression vector pQE30. Polypeptides encoded by these fragments were shown to be synthesized in Escherichia coli with a yield of up to 100 micrograms/ml of culture and to be accumulated in the cells as inclusion bodies. The partially purified antigens were used in enzyme immunoassay with the sera of humans convalescing from epidemic typhus, tick-borne rickettsioses, and other infectious diseases. One of two recombinant proteins was shown to react in immunoblotting and ELISA with homologous, but not with heterologous, sera. The immunoreactivities in ELISA of the recombinant antigens and heat-denatured SPA proved to be similar, but substantially lower than that of the native SPA. These data as well as the data of other investigators show that serodiagnostics of epidemic typhus using recombinant antigens remains a problem. PMID- 11040998 TI - [Effect of surface-active agents (SAA) on peroxidase activity. III. Effect of nonionic SAA]. AB - The effect of a series of nonionic surfactants on the initial rate of the peroxide oxidation of 5-aminosalicylic acid in solution catalyzed by horseradish peroxidase was studied. As the surfactant concentration increases, the peroxidation rate first increases, then decreases, and the increase/decrease cycle is repeated. The primary increase may be induced by a change in properties of the medium under the action of surfactants, and the following decrease, by the enzyme inhibition. The secondary increase may be explained by a change in the enzyme conformation and an increase in the accessibility of its active site for the substrate due to the immobilization of the protein in the surfactant aggregates, whereas the secondary decrease, by a shielding of the protein with these aggregates. PMID- 11040997 TI - [Identification of Escherichia coli nitrate reductase as an antigen for a monoclonal antibody with previously unknown specificity]. AB - The immunoaffinity chromatography of total membrane proteins from Escherichia coli helped determine the specificity of the monoclonal antibody 3A6 that was obtained upon immunization of mice with nicotinamide nucleotide transhydrogenase preparations and reacted with an unknown E. coli antigen. Proteins with apparent molecular masses of 150, 45, and 20 kDa were isolated and identified by N terminal sequencing as the subunits of nitrate reductase. This conclusion was confirmed by immunoblotting with the 3A6 antibody of the proteins from the E. coli cells grown upon induction of nitrate reductase. It was shown that the 3A6 antibody specifically recognizes the alpha subunit of nitrate reductase, and the formation of the enzyme-antibody complex does not result in a loss of the enzyme catalytic activity. PMID- 11040999 TI - [Effect of surface-active agents on peroxidase activity. IV. Effect of lipids and fatty acids from milk]. AB - The effect of surfactants, lipids and fatty acid salts isolated from cow milk on the activity of heme-containing horseradish peroxidase in solution was studied. As the surfactant concentration increases, the rate of the enzymic reaction successively decreases, increases, and again decreases, down to zero in the case of the fatty acid salts. The initial deceleration of the reaction rate results from the enzyme inhibition. The subsequent increase is caused by an improved accessibility for the substrate and the enhanced activity of the catalytic site of the enzyme due to its immobilization in the surfactant aggregates. A shielding of the protein by these aggregates can explain the secondary deceleration of the enzymic reaction rate. The general character of the dependence is similar and does not depend on the surfactant structure for a series of fatty acid salts and phospholipids; however, it is quite different in the case of cholesterol and sphingomyelin. PMID- 11041000 TI - [Production of beta-xylosidase by the yeast Cryptococcus podzolicus]. AB - In studying the beta-xylosidase production by yeast Cryptococcus podzolicus (Basidiomycetes), it was shown to be an inducible secretory enzyme. Xylooligosaccharides generated from xylan and methyl beta-xyloside were found to induce the beta-xylosidase synthesis. The enzyme activity in the medium containing xylan or methyl beta-xyloside was 1.0 and 1.5 U/ml, respectively; this production level is similar to that achievable at the beta-xylosidase production by mycelial fungi. PMID- 11041001 TI - [Sensitized photomodification of DNA with binary sysytems of oligonucleotide conjugates. VI. Effect of substituents in the anthracene residue of the sensitizer]. AB - Photomodification of ssDNA by binary systems of oligonucleotide conjugates complementary to the adjacent sequences of the target DNA was studied. One of the conjugates comprised a substituted anthracene as a sensitizer; the other, p azidotetrafluorobenzaldehyde 3-aminopropionylhydrazone as a photoreagent. The sensitized photomodification is initiated by the 365-580-nm light through an efficient energy transfer from the photoexcitated sensitizer onto the photoreagent in a complementary complex of the binary system with the DNA target where the sensitizer and the photoreagent are sterically converged. Influence of substituents in the anthracene residue on the efficiency of the DNA sensitized photomodification was considered. The oligonucleotide conjugate of anthracene-9 al 3-aminopropionylhydrazone allows highly specific initiation of the sensitized photomodification upon irradiation with visible light at > 460 nm in conditions generating no photoreaction in the sensitizer's absence. PMID- 11041002 TI - [Chromosomal localization of rpb9+ and tfa1+ genes, coding for components of the mRNA synthesis apparatus of Schizosaccharomyces pombe]. AB - Using DNA hybridization on cosmid filters of high density, we established chromosomal localization of the rpb9+ gene encoding one of the specific subunits of RNA polymerase II of Schizosaccharomyces pombe and thus filled in the last gap in the mapping of the genes encoding components of RNA polymerase II of the fission yeast. The primary structure of three extended regions of the Sz. pombe chromosome I was elucidated and, as a result, genes neighboring on rpb9+ were identified. One of them proved to be the tfa1+ gene, encoding the large (alpha) subunit of the general factor of transcription initiation TFIIE. PMID- 11041003 TI - [Study of dimerization of polysynthetic derivatives of the antibiotic eremomycin by ESI MS and its role in elucidating antibacterial activity]. AB - The dimerization constants for glycopeptide antibiotics vancomycin, ristocetin, and eremomycin and nine semisynthetic eremomycin derivatives were determined by the electrospray ionization mass spectrometry; the constants for natural antibiotics turned out to be close to those previously determined by NMR. No correlation between these dimerization constants and antibacterial activities of all the compounds toward the clinical strains of Gram-positive bacteria was found. PMID- 11041004 TI - 'I couldn't have known': accountability, foreseeability and counterfactual denials of responsibility. AB - This article explores situational determinants and psychological consequences of 'counterfactual excuse-making'--denying responsibility by declaring 'I couldn't have known...'. Participants who were made accountable for a stock investment decision that resulted in an outcome caused by unforeseeable circumstances were particularly likely to generate counterfactual excuses and, as a result, to deny responsibility for the outcome of their choices and minimize their perceptions of control over the decision process. The article discusses the implications of these findings for structuring accountability reporting relationships in business and, more generally, stresses the benefits of counterfactual denials of responsibility for maintaining self-esteem and a desired self-identity. PMID- 11041005 TI - Simulation versus the thing itself: commentary on Markman and Tetlock. PMID- 11041006 TI - 'Going to where the action is' versus 'creating the action': a reply to Antaki. PMID- 11041007 TI - Attitude-behaviour relations: the role of in-group norms and mode of behavioural decision-making. AB - Two experiments provided support for the central hypothesis--derived from social identity/self-categorization theories--that attitudes would be most likely to predict behaviour when they were supported by a congruent in-group norm. In the first experiment, norm congruency and mode of behavioural decision-making (spontaneous or deliberative) were orthogonally manipulated in a between-subjects study of career choice in psychology. Participants exposed to an attitudinally congruent in-group norm towards their preferred career choice were more likely to display attitude-behaviour consistency than those exposed to an attitudinally inconsistent group norm, an effect that was evident under both spontaneous and deliberative decision-making conditions. Using a mock jury paradigm, Expt 2 replicated and extended the first experiment by including a manipulation of in group salience. As predicted, participants exposed to an incongruent norm displayed greater attitude-behaviour inconsistency than those exposed to a congruent norm. Contrary to predictions, this effect did not vary as a function of group salience, nor did the effects of group norms for high and low salience participants vary as a function of mode of behavioural decision-making. However, there was evidence that perceived identification with the group moderated the influence of norms on attitude-behaviour consistency. PMID- 11041008 TI - The structure of attitudes: attribute importance, accessibility and judgment. AB - Two studies related attribute importance to accessibility and speed of judgment. Attitudes were assessed by a direct attitude measure and a modal set of 15 attributes. Attributes were rated in terms of their probability and desirability. Subsequently, participants were required to select the five attributes they considered to be most important. Results of Study 1 (dealing with attitudes towards condom use) show that individually selected, important attributes are more easily retrieved and judged faster than non-selected, less important attributes. Judging attributes took less time than evaluating one's overall attitude, suggesting a bottom-up process in which the various attributes are combined to form an overall attitude. Study 2 (dealing with attitudes towards smoking) extends these findings and also addresses the stability of attitude structure. Important attributes were again associated with reduced response times, and attribute-related judgments took less time than judging one's overall attitude. Accessibility of important attributes remained stable over time as indicated by the results of a lexical decision task one week later. Finally, important attributes were also recalled better than less important attributes. Implications for research on attitude structure are discussed. PMID- 11041009 TI - 'But I'm different to them': constructing contrasts between self and others in talk-in-interaction. AB - Contrast occurs as an important theme across a range of post-structural, conversation and discourse analytic and 'traditional' social psychological work. Some approaches link contrasts to identity issues--thus self-categorization theory argues that identity arises in part through self assignment to particular categories--which is done on the basis of one's perception of similarities and differences with others. This paper explores contrast in identity talk from a different vantage point: one which has the strengths and shortcomings of an emphasis upon talk in interaction rather than on cognition in isolation. In so doing it draws upon a corpus of extracts largely derived from unstructured interviews with television news audiences but supplemented with some published data of relevance to the deployment of contrast with others. The paper illustrates some of the different ways in which contrasts with others can be constructed and deployed, which may challenge a category assignment, construct a generic category of others against whom self is favourably positioned, or act as an interactionally astute self-deprecation. In each case the contrast is understood as tuned in to the interactional issues at hand. In this way contrast is conceptualized as a live talk-in-interaction activity which can be fruitfully explored from the perspective of the participants themselves. PMID- 11041010 TI - Pattern of disconfirming information and processing instructions as determinants of stereotype change. AB - This experiment examined the effects of pattern of disconfirming information (concentrated vs. dispersed) and processing instructions (focus on similarities vs. differences vs. control) on stereotype change. If subtyping and perceived typicality are central to the stereotype change process, then processing instructions designed to affect these processes should affect stereotyping. There was lower stereotyping when perceivers focused on similarities between group members, and after exposure to a dispersed pattern of disconfirming information. Only the main effect of pattern was mediated by the perceived typicality of disconfirmers, but not by an index of subtyping based on clustering of information from disconfirmers in recall. Results support a model of stereotype change in terms of the impact of disconfirming group members who are also seen as typical of the group; subtyping of extreme disconfirmers may work in parallel, or later, and contribute to the long-term maintenance of a stereotype. PMID- 11041011 TI - Context effects of facial appearance on attitudes toward mentally handicapped persons. AB - The influence of facial appearance on social attitudes was examined by exposing participants to the faces of three target persons with or without deviant facial features, posing happy, angry or sad facial expressions, or a mixture of these expressions. When they displayed negative emotional expressions, facially deviant targets were judged more negatively than non-deviant targets. Irrespective of emotional expression and level of personal experience, participants expressed more negative attitudes toward mentally handicapped persons in general after exposure to deviant faces than after exposure to non-deviant faces, or in the absence of exposure. However, correlational analyses suggested that only at low levels of personal experience were attitudes influenced by previously formed impressions of deviant exemplars. Results are discussed in terms of the motivational relevance of physical features in stigmatization, and context and exemplar effects in stereotyping and attitude measurement. Practical implications are also discussed. PMID- 11041012 TI - Causal attribution and Mill's methods of experimental inquiry: past, present and prospect. AB - J. S. Mill proposed a set of Methods of Experimental Inquiry that were intended to guide causal inference under every conceivable set of circumstances in which experiments or observations could be carried out. The conceptual and historical relationship between these Methods and modern models of causal attribution is investigated. Mill's work retains contemporary relevance because his insights show how research can progress into presently uncharted waters. Following Mill, it is proposed that people use many different methods of causal attribution, the nature of which remains to be ascertained, and that the conditions that affect choice of method include the need to eliminate alternative causal candidates, whether single or multiple events are to be explained, the use of intervention or experiment as opposed to mere observation, and practical concerns. PMID- 11041013 TI - Is racism dead? Comparing (expressive) means and (structural equation) models. AB - Much scholarship suggests that racism--belief in out-group inferiority--is unrelated to contemporary attitudes. Purportedly, a new form of racism, one which relies upon a belief in cultural difference, has become a more acceptable basis for such attitudes. The authors argue that an appropriate empirical assessment of racism (both 'old' and 'new') depends upon (1) clear conceptualization and operationalization, and (2) attention to both mean-level expression and explanatory value in structural equation models. This study assessed the endorsement of racism and belief in cultural difference as well as their association with a measure of general attitude in a secondary analysis of parallel representative surveys of attitudes toward different ethnic out-groups in France, The Netherlands, Western Germany and Britain (N = 3242; see Reif & Melich, 1991). For six of the seven out-group targets, racism was strongly related to ethnic majority attitudes, despite low mean-level endorsement. In a pattern consistent with a 'new', indirect racism, the relationship between British racism and attitudes toward Afro-Caribbeans was mediated by belief in cultural difference. PMID- 11041014 TI - Clinical results and the necessity of estimating patient-specific radiation absorbed dose in radioimmunotherapy. PMID- 11041016 TI - Antibody therapy of acute myelogenous leukemia. AB - Monoclonal antibodies (mAbs) have become an important modality for cancer therapy. A genetically engineered, humanized anti-CD33 antibody HuM195 has demonstrated activity against over relapsed acute myelogenous leukemia (AML) and against minimal residual disease in acute promyelocytic leukemia (APL). Radioimmunotherapy with beta (beta) particle-emitting isotopes has produced significant responses while minimizing radiation exposure to normal tissues in both nonmyeloablative and myeloablative regimens. Targeted alpha (alpha) particle therapy with 213Bi-labeled HuM195 offers the possibility of more selective tumor cell kill. Additionally, directed chemotherapy using an anti-CD33-calicheamicin conjugate (CMA-676) has produced remissions in patients with relapsed AML. PMID- 11041015 TI - Radiopharmaceuticals for nuclear endocrinology at the University of Michigan. AB - The historical background at the University of Michigan laid a foundation for the innovative development of radionuclides in diagnosis and treatment of endocrine diseases. From that background, Dr. William Beierwaltes, the chief of Nuclear Medicine, inspired two talented young chemists to synthesize unique radiopharmaceuticals that transformed diagnostic approaches to certain endocrine disorders. Dr. Raymond Counsell's 131-I-radiocholesterol, enabled imaging that defined function in the adrenal cortex, and thereby distinguished the different forms of Cushing's syndrome and of primary aldosteronism; in addition, this new technique differentiated benign adrenal cortical adenomas from other adrenal cortical tumors. Dr. Donald Wieland created metaiodobenzlylguanidine (MIBG), a compound that can be tagged with either 131-I or 123-I, and led to the scintigraphic depiction of adrenergic tumors, particularly pheochromocytomas and neuroblastoma, anywhere in the body of a patient. Treatments with large doses of MIBG have reduced the malignant forms of pheochromocytomas and brought remissions to children with neuroblastomas. MIBG also concentrated in the autonomic neurons and so the nerves of the heart were also portrayed. Subsequent novel syntheses included positron-emitting nuclides that, through positron emission tomography, have revealed the physiology and altered physiology of the human heart. These men and their discoveries exemplify the creative endeavors that compel us to seek further the wonders of nuclear science. PMID- 11041017 TI - Scintigraphic detection of multidrug resistance in cancer. AB - One of the most extensively studied mechanisms of drug resistance involves the drug efflux pump P-glycoprotein (Pgp). The availability of radiolabeled substrates of Pgp such as 99mTc-MIBI and analogous 99mTc-labeled agents allows the clinical assessment of Pgp function in cancer patients. The consistency of the results from different institutions and trials strongly support the clinical application of this imaging technique for individual tailoring of chemotherapeutic regimens and for designing clinical trials with Pgp modulators. PMID- 11041018 TI - In vitro comparison of sestamibi, tetrofosmin, and furifosmin as agents for functional imaging of multidrug resistance in tumors. AB - Sestamibi, tetrofosmin, and furifosmin are 99mTc-labeled myocardial perfusion imaging agents which have been shown to be substrates for P-glycoprotein (Pgp), the multidrug-resistance transporter which is overexpressed in some tumors. The three tracers were directly compared in vitro in the human breast cancer cell line MCF7-WT and two multidrug-resistant variants, MCF7-BC19 (MDR1 gene transfected) and MCF7-AdrR (doxorubicin selected). Tracer accumulation over the course of 60 minutes was determined. Dose-response curves were generated for two modulators of Pgp function, GG918 and PSC833. The general shape of accumulation curves for the three tracers in MCF7-WT cells was similar, with accumulation levels being sestamibi > tetrofosmin > furifosmin. Accumulation of sestamibi and furifosmin in MCF7-BC19 cells was reduced to 10% and 21% of MCF7-WT levels, respectively, but this accumulation deficit could be completely reversed by addition of 0.1 microM GG918 or 2 microM PSC833. Accumulation of sestamibi and tetrofosmin in MCF7-AdrR cells was 1.6% and 12% of MCF7-WT levels, respectively, and could only be enhanced to 30% and 45% of MCF7-WT levels by addition of GG918 or PSC833. In contrast, furifosmin showed similar levels of accumulation in MCF7 WT and MCF7-BC19 cells, slightly lower levels in MCF7-AdrR cells, and no consistent response to Pgp modulators. These results support the continued investigation of sestamibi and tetrofosmin as agents for functional imaging of multidrug resistance in human cancer. PMID- 11041019 TI - Tumor-absorbed-dose estimates versus response in tositumomab therapy of previously untreated patients with follicular non-Hodgkin's lymphoma: preliminary report. AB - I-131-radiolabeled tositumomab (Anti-B1 Antibody), in conjunction with unlabeled tositumomab, was employed in a phase II clinical trial for the therapy of 76 previously-untreated follicular-non-Hodgkin's-lymphoma patients at the University of Michigan Cancer Center. For all patients, conjugate-view images were obtained at six to eight time points on seven consecutive days after a tracer infusion of the antibody. A SPECT image set was obtained on day two or three after the therapy infusion for 57 of the patients. Of these, 55 are suitable for dosimetric evaluation. To date, we have completed analysis and response characterization of 20 patients from the subset of 55. All 20 patients had either a complete response (CR) or a partial response (PR). Conjugate-views provided a time-activity curve for a composite of nearby, individual tumors. These tumors were unresolved in the anterior-posterior projection. Pre-therapy CT provided volume estimates. Therapy radiation dose was computed for the composite tumor by standard MIRD methods. Intra-therapy SPECT allowed the calculation of a separate dose estimate for each individual tumor associated with the composite tumor. Average dose estimates for each patient were also calculated. The 30 individual tumors in PR patients had a mean radiation dose of (369 +/- 54) cGy, while the 56 individual tumors in CR patients had a mean radiation dose of (720 +/- 80) cGy. According to a mixed ANOVA analysis, there was a trend toward a significant difference between the radiation dose absorbed by individual tumors for PR patients and that for CR patients. When the radiation dose depended on only the patient response, the p value was 0.04. When the radiation dose depended on the pre-therapy volume of the individual tumor as well as on the patient response, the p value was 0.06. Since the patient response was complete in 75% of the patients, the analysis of the total cohort of 55 evaluable patients is needed to have a larger number of PR patients to better test the trend toward a significant difference. A pseudo prediction analysis for patient-level dose and response was also carried out. The positive predictive value and the negative predictive value were 73% and 80%, respectively when a patient's average radiation dose was used. The predictive values were 73% and 60%, respectively, when the patient's average base-10 logarithm of radiation dose was used. A complete overlap for the dose range of CR patients compared to that for PR patients precluded higher predictive values. In conclusion, there was a trend toward a significant difference in the radiation dose between CR and PR patients, but it was only moderately predictive of response. PMID- 11041020 TI - Growth of tumor-derived activated T cells for the treatment of advanced cancer. AB - In 1994, we reported on a series of patients treated with T-cell therapy (Study #1). This paper (Study #2) is an update of our experience through 1999 in the production of tumor-derived activated cells (TDAC), also called tumor infiltrating lymphocytes (TIL), from tumor biopsies. TDAC were successfully grown in medium containing Interleukin-2 from 75% of the 366 tumor biopsies tested. There was no significant difference in success (growth to 1 x 10(9) cells) comparing primary and metastatic tumors. Many of the tumors were shipped to the laboratory by overnight delivery from distant sites. Success rate did decrease with the length of time for tumor transport in excess of 24 hours. Certain additional cytokines were tested when cultures did not grow. Interleukin-4 was beneficial in the development of 1 of 4 TDAC cultures which did not grow with IL 2 alone. In order to produce TDAC to treat patients, cells were grown in gas permeable plastic bags or in artificial capillary bioreactor cultures. Approximately 1 x 10(9) were seeded from an initially successful "feasibility study" to bulk produce cells for treatment. Harvest was carried out after about 3 weeks. Sixty-three patients were treated at least once with a minimum of 1 x 10(10) TDAC given by intravenous infusion. On the average, the number of cells per treatment was 3 x 10(10) with a viability of 87%. TDAC cultures contained T cells with variable ratios of CD4 to CD8 cells. Secreted granulocyte-monocyte colony stimulating factor, interferon gamma and tumor necrosis factor alpha were measured in TDAC conditioned medium. Only 34 patients received the full course of 4 TDAC treatments. The cells were well tolerated with mild fever and dyspnea. Partial responses were observed in 8 patients, including the dramatic regression of scalp nodules in a patient with renal cancer. These results showed that therapeutic amounts of TDAC can be produced in cell culture in a reasonable and cost-effective manner. The cells were well tolerated and responses were seen in renal and melanoma patients resistant to IL-2 with bulky, advanced cancer. PMID- 11041021 TI - Tc-99m MIBI in suspected recurrent breast cancer. AB - A prospective trial was performed to assess the accuracy of Tc-99m MIBI scintimammography in 63 women (mean age 65, range 33-85 years) with suspected recurrent breast cancer in the breast and/or loco-regional tissues. All patients had been diagnosed with breast cancer 1-23 years before the scintimammography. A total of 27 breasts had been removed by mastectomy so scintimammography was compared with mammography in the remaining 99 breasts. Pathological follow-up of patients confirmed 33 sites of recurrent disease within the breast, 26 (78%) were identified by scintimammography and 14 (42%) by mammography; 30 (90%) were positive on one test or the other. In addition Tc-99m MIBI scintimammography identified 10/16 (63%) of axillary lymph nodes with recurrent tumour and 4/6 sites of recurrent tumour present elsewhere. Tc-99m MIBI scintimammography is more accurate in identifying recurrent disease in the breast than mammography and can identify loco-regional recurrence outside of the breast. PMID- 11041022 TI - Optimal timing of administration of hyperthermia in combined radioimmunotherapy. AB - Local hyperthermia (HT) may enhance the efficacy of radioimmunotherapy (RIT). However, the optimal timing of HT relative to administration of antibody is unknown. Human colon cancer xenografts (290 +/- 26 mm3) were treated with 4.63 MBq 131I-A7 monoclonal antibody (MAb) anti-Mr 45,000 glycoprotein antigen on colorectal cancer, and HT at 43 degrees C for 1 h was administered at: (A), 2 days after the 131I-A7 injection at the maximum 131I-A7 tumor accumulation (radiation); (B), soon after the 131I-A7 injection aiming to increase the tumor accumulation of 131I-A7 due to HT vascular effects; or (C), 2 days before the 131I-A7 injection in an attempt at injecting 131I-A7 when increased antigen expression could be expected. Specific growth delay (SGD) of tumors was calculated as (Tqtreat-Tqcontrol)/Tqcontrol where Tq was tumor quadrupling time. The biodistribution and intratumoral distribution of 131I-A7 were investigated to explore the mechanism of tumor response among the different HT regimens. HT alone produced some antitumor effect (SGD 1.90 +/- 0.26), which was less effective than RIT (3.11 +/- 0.50). HT soon after 131I-A7 RIT (B) significantly enhanced RIT efficacy (6.57 +/- 0.51, p < 0.0001) whereas neither HT at 2 days after RIT (A) nor at 2 days before RIT (C) did so. Biodistribution study revealed that HT soon after RIT (B) increased the tumor radiation absorbed dose by a factor of 2.4, while HT after RIT (A) did not increase radiation dose and HT before RIT (C) decreased it. Radioluminograms of tumor sections indicated that HT soon after RIT (B) improved the uniformity of 131I-A7 distribution whereas HT after RIT (A) did not and HT before RIT (C) diminished the uniformity of A7 distribution. In conclusion, the best therapeutic efficacy was obtained when HT was combined soon after the initiation of RIT with 131I-A7. The increased tumor radiation absorbed dose and the uniform intratumoral distribution of 131I-A7 were important factors underlying this improvement, and the additive cytotoxicity of HT is suspected to some extent. HT-induced radiosensitization of tumor was not apparent in this model when HT was given 2 days after 131I-A7 MAb. PMID- 11041023 TI - Comparison of Tc-99m sestamibi, serum neuron-specific enolase and lactate dehydrogenase as predictors of response to chemotherapy in small cell lung cancer. AB - Neuron-specific enolase (NSE) and lactate dehydrogenase (LDH) are tumor markers of small cell lung cancer (SCLC) which were reported to predict outcome of patients with SCLC. We previously reported that dipyridamole-modulated Tc-99m sestamibi (dipyridamole-MIBI) single photon emission computed tomography (SPECT) could predict the response to chemotherapy in SCLC patients. The purpose of this study was to compare dipyridamole-MIBI and pretreatment serum levels of NSE and LDH for the prediction of response to chemotherapy in SCLC. Twenty-eight SCLC patients underwent dipyridamole-MIBI SPECT 3 to 7 days before starting chemotherapy (80 mg/m2 etoposide and 80 mg/m2 cisplatin every 3 or 4 weeks for at lease two cycles). Serum levels of NSE and LDH were also measured at the same day of the imaging. Tomographic images before and after 0.84 mg/kg dipyridamole infusion were acquired 1 hour after injection of 370 (10 mCi) and 1,110 (30 mCi) MBq MIBI, respectively. The response to chemotherapy was grouped as specified as complete (CR), partial response (PR), no change (NC), and progressive disease (PD), according to the change in tumor size on chest roentgenography and CT. Patients showing CR and PR were classified as responders, and those who showed NC and PD were considered nonresponders. Among the 28 patients, 15 were responders (2 CR, 13 PR) and 13 were nonresponders (11 NC, 2 PD). The change of tumor-to normal lung ratio (T:NL) after infusion of dipyridamole was significantly higher in responders as compared with nonresponders (0.38 +/- 0.64 vs. -0.38 +/- 0.50, respectively, p = 0.002). However, pretreatment serum NSE and LDH levels did not correlate with the response to chemotherapy. Increase of T:NL after dipyridamole infusion was a strong negative predictor of chemotherapeutic response in SCLC patients, while NSE and LDH could not predict it. PMID- 11041024 TI - Sudden infant death syndrome in 2000. PMID- 11041025 TI - Evolutionary anticipation of the human heart. AB - We have studied the comparative anatomy of hearts from fish, frog, turtle, snake, crocodile, birds (duck, chicken, quail), mammals (elephant, dolphin, sheep, goat, ox, baboon, wallaby, mouse, rabbit, possum, echidna) and man. The findings were analysed with respect to the mechanism of evolution of the heart. PMID- 11041026 TI - Anatomical basis for impotence following haemorrhoid sclerotherapy. AB - Impotence has been reported as a rare but important complication of sclerotherapy for haemorrhoids. The relationship between the anterior wall of the rectum and the periprostatic parasympathetic nerves responsible for penile erection was studied to investigate a potential anatomical explanation for this therapeutic complication. A tissue block containing the anal canal, rectum and prostate was removed from each of six male cadaveric subjects. The dimensions of the components of the rectal wall and the distance between the rectal lumen and parasympathetic nerves in the periprostatic plexus were measured in horizontal transverse histological sections of the tissue blocks at the level of the lower prostate gland (i.e. the correct level for sclerosant injection). The correct site of sclerosant in the submucosa was on average 0.6 mm (SD 0.3 mm) deep to the rectal mucosal surface and only 0.7 mm (SD 0.5 mm) in thickness. The nearest parasympathetic ganglion cells were a mean of only 8.1 mm (SD 2.0 mm) deep to the rectal lumen. The close proximity of the rectum to the periprostatic parasympathetic nerves defines an anatomical basis for impotence following sclerotherapy. This emphasises the need for all practitioners to be particularly careful when injecting in this area and for strict supervision of trainees. PMID- 11041027 TI - Evaluation of pressure beneath a split above elbow plaster cast. AB - It has previously been shown that splitting a plaster cast after manipulation of, or surgery on, a limb leads to a decrease in pressure beneath the cast by accommodating the swelling that may occur. However, it is not known whether the axis along which the cast is split influences the amount of swelling that can occur before a critical pressure is reached. We investigated this with reference to above elbow plaster casts. PMID- 11041028 TI - Endoscopic ultrasound in the staging of tumours of the oesophagus and gastro oesophageal junction. AB - BACKGROUND: Modern management of upper gastro-intestinal cancer demands accurate pre-operative staging. In continental Europe and Japan, endoscopic ultrasound (EUS) is established as the investigation of choice for local staging of these cancers, but British experience with this technique is limited. METHODS: A retrospective review of the medical records of patients with oesophageal or gastro-oesophageal junction tumours during our first 3.5 years' experience with EUS was undertaken and the findings at EUS correlated with the pathology of the resected specimen. RESULTS: A total of 124 patients (86 males), with a mean age of 64.5 years, underwent EUS: 84 had adenocarcinoma and 26 squamous cell carcinoma. There were 3 failed EUS examinations, 42 patients did not have surgery for a variety of reasons, and 10 patients had pre-operative chemoradiotherapy. In the remaining 69 patients, correlation for T stage showed an accuracy of EUS of 80% and for N staging of 54% overall. Comparison of the initial 2 years with the final 18 months showed no change in the T staging accuracy but an improvement in the N staging accuracy from 50% to 60%. CONCLUSION: Once initial experience has been gained, EUS is an accurate procedure for T and N staging of tumours of the oesophagus and gastro-oesophageal junction. It should be included with other imaging modalities, such as CT scanning, in the pre-operative assessment of these tumours. PMID- 11041029 TI - Shielding reproductive organs of orthopaedic patients during pelvic radiography. AB - The use of gonadal shielding has been advocated for patients undergoing pelvic radiography before and during the reproductive years. The aim of this study is to look at the adequacy of gonadal shielding used in a district general hospital for young patients having pelvic radiographs. A total of 200 radiographs were reviewed of 49 patients below the age of 45 years. Full coverage was achieved in only 36% of cases. Amongst females, only 22% received adequate shielding. None of the patients in their reproductive years (16-45 years) had gonad shields. The reasons for inadequate coverage were, in order of frequency: (i) no shielding was used; (ii) malposition of the shielding device; and (iii) the use of inappropriately shaped or sized devices. Suggestions for improvement are proposed. PMID- 11041030 TI - Delays in orthopaedic trauma treatment: setting standards for the time interval between admission and operation. AB - Delay in operating on trauma patients leads to increased morbidity, mortality, length of hospital stay and overall cost. The urgency of operative intervention depends on the injury sustained. There are no published guidelines on what constitutes a reasonable delay between admission and operation. As part of the clinical governance in our unit, an audit was undertaken to examine the structure and process of trauma operating. Patients were allocated to groups defined by the Bath Orthopaedic Department, according to urgency of need for surgery. Group A: patients (for example, open fractures and dislocations) should have definitive treatment within 6 h of admission. Group B: patients (for example, hip fractures, long bone injuries and ankle fractures) should have operations on the day that they are presented to the consultant trauma meeting, or on the day that they are declared fit/ready for theatre. Group C: patients (for example, tendon injuries, simple hand fractures) should have operations within 5 days of presentation to the trauma meeting. Over 3 months, there were 401 acute orthopaedic admissions requiring surgery (61 group A, 277 group B, 63 group C). 78% of group A patients, 58% of group B patients and 86% of group C patients were operated on within the target times. In total, 137 out of 401 operations (34%) missed the targets set. 119 of these (87%) were delayed due to lack of available operating time. This was despite the fact that 59 operations (15% of total) were done on lists normally used for elective operating. Most of the other delays were due to the need for an appropriately experienced surgeon to be available. If these targets are to be achieved for the majority of patients, the trauma theatre must become more efficient, or more flexible time must be made available during evenings or weekends to clear the backlog of trauma operations. PMID- 11041031 TI - Unexpected overnight admissions following day-case surgery: an analysis of a dedicated ENT day care unit. AB - Day-case surgery is an integral part of otolaryngology, and many procedures can be performed as day-cases provided strict criteria are applied in the selection of patients. We reviewed patients who required unexpected admission from the day case unit at the Royal National Throat, Nose and Ear Hospital, London between April 1997 and March 1998. The total number of patients undergoing surgery was 1642. Of the total, 29 (1.8%) had to be admitted unexpectedly for overnight stay: 24 of these patients had undergone nasal surgery, representing 5.4% of all the nasal procedures performed--and the cause of all these admissions was haemorrhage. Further analysis revealed 22 of these 24 nasal operations had included a septoplasty. The total number of septoplasties performed was 163; thus, septoplasty had an unexpected admission rate of 13.4%. This information has been used to formulate stricter guidelines for day-case septoplasty admissions in our unit. PMID- 11041032 TI - The nurse practitioner endoscopist. AB - Most upper and lower gastrointestinal endoscopies in Great Britain and Ireland are performed by surgeons, physicians or radiologists. Since the introduction of the 'nurse endoscopist' by the British Society of Gastroenterology Working Party, few centres in the UK have adopted this policy. We have reviewed the anxiety about nurse practitioner endoscopists among patients and physicians. Finally, the role and future of the nurse practitioner endoscopist in the UK is discussed. PMID- 11041033 TI - A 23-year review of the management of acute retention of urine: progressing or regressing? AB - A retrospective review of all patients in Oxford under the care of one consultant urologist (GJF) who presented on alternate years over a 23-year period with acute retention of urine was undertaken. Data were collected on the: (i) number of patients discharged from hospital with an in-dwelling catheter; (ii) duration of catheter drainage prior to surgery; and (iii) duration of postoperative stay. In all, 244 patients underwent prostatectomy. Over the 23-year period, there was a significant increase in the proportion of patients discharged prior to surgery (P < 0.001) as well as their median duration of catheterisation (P < 0.001): more than 50% were catheterised for more than 3 months in 1997. Conversely, post operative hospital stay has decreased. Prolonged catheter drainage carries considerable morbidity, with 72% experiencing some complication. Most patients feel they lose dignity, 69% consider it uncomfortable and more than 50% complain of burning sensations, bladder spasms and a persistent desire to micturate. We recommend that patients should not be placed on routine waiting lists where they are liable to remain for an unacceptably long time. Targets should be set to admit them within a set period and theatre lists made available. We feel that six weeks is a realistic target. PMID- 11041034 TI - The influence of experience and specialisation on the reliability of a common clinical sign. AB - OBJECTIVES: To explore the influence of experience and specialisation on clinical judgement by comparing accuracy in diagnosing anaemia between a consultant general surgeon, a consultant ophthalmologist and their registrars. PATIENTS AND METHODS: Conjunctival inspection of 101 patients, subsequent correlation with haemoglobin concentration. MAIN OUTCOME MEASURES: Number of correct and incorrect diagnoses of anaemia. RESULTS: 54 patients were anaemic and 47 were not. Overall accuracy in diagnosing anaemia ranged from 0.61-0.69, sensitivity 0.52-0.65 and specificity 0.62-0.83. Agreements between pairs of examiners were 0.68-0.81, with kappa values of 0.36-0.60 when adjusted for chance agreement. CONCLUSIONS: Neither experience nor specialisation significantly influenced our ability to diagnose anaemia, based on conjunctival inspection. Without critical analysis of clinical signs, we are unaware of their diagnostic limitations. PMID- 11041035 TI - Harvesting split skin grafts of appropriate thickness using the hand-held knife. AB - Split skin grafting is an often used method of covering small skin defects and ulcers. We describe a simple technique to help determine the appropriate setting for the hand-held knife. This provides skin of adequate thickness when harvesting graft from the thigh and permits good regeneration of the donor site. PMID- 11041037 TI - Testing for anastomotic integrity after reversal of loop ileostomy. AB - A technique is described to test the integrity of the anastomosis formed during reversal of ileostomy intra-operatively and the early clinical results obtained using the technique. PMID- 11041036 TI - Surgical management of infantile hypertrophic pyloric stenosis--can it be performed by general surgeons? AB - INTRODUCTION: Debate exists as to whether IHPS can be treated in district general hospitals as effectively as in specialist paediatric surgical units. AIM: To review the surgical treatment of IHPS in babies admitted to a district general hospital under the care of two consultant general surgeons with a paediatric surgical interest. PATIENTS AND METHODS: The case notes of 66 babies operated on for IHPS over a 42 month period between April 1995 and September 1998 were retrospectively reviewed. Demographics, operative details, hospital stay, and overall complications were all documented. RESULTS: Peri-operative complications occurred in 2 patients, both requiring omental patches for duodenal perforation. Nine patients had 1 or 2 episodes of postoperative vomiting; 4 had either a wound or urinary tract infection; and 1 baby developed an incisional hernia. There was no mortality. DISCUSSION: The complication rate seen in this series is comparable to that of specialist centres, and supports current guidelines suggesting that IHPS can be managed by general surgeons with a paediatric surgical interest in a district general hospital. PMID- 11041038 TI - Wound irrigation: a simple, reproducible device. AB - It is well established that copious saline irrigation is essential to good wound toilet with low rates of contamination. We present a simple and inexpensive method of wound irrigation that is ideal for use in both casualty and theatre situations. PMID- 11041039 TI - Postoperative minocycline pigmentation. AB - This paper describes an unusual case of florid postoperative pigmentation caused by minocycline which was not diagnosed for some 8 months, causing anxiety and distress to both patient and surgeon. PMID- 11041040 TI - Pneumoperitoneum following Jacuzzi usage. AB - A 56-year-old woman presented with abdominal pain after using a Jacuzzi hours earlier. Abdominal radiographs revealed intra-peritoneal free gas and, as she presented symptomatically, a laparotomy was performed. This revealed fluid and gas but no visceral perforation or intra-abdominal pathology to account for this. Peritoneal lavage was performed and the patient made an unremarkable recovery. Various causes of pneumoperitoneum have been described in the literature and both conservative and operative treatment recommended. We are unaware of any other reports of Jacuzzi-induced pneumoperitoneum and describe it as an entity to be considered in abdominal pain secondary to the use of similar types of device. PMID- 11041041 TI - Pyomyositis mimicking right iliac fossa mass: review of the literature. AB - Pyomyositis is a pyogenic infection of skeletal muscle. Its incidence in temperate countries though low is rising. Most cases from the temperate region involve immuno-compromised patients. The onset is usually insidious with progression to large purulent collections. Because of its low incidence in temperate countries, it is often initially misdiagnosed. A high index of suspicion with appropriate imaging techniques, aggressive surgical intervention and adjunctive antibiotic therapy are the keys to prompt resolution. A case of pyomyositis mimicking right iliac fossa (RIF) mass is described with a review of the literature. PMID- 11041042 TI - An unusual cause of femoral embolism: angioseal. PMID- 11041043 TI - How to draw the ellipse of skin for a mastectomy. PMID- 11041044 TI - How to draw the ellipse of skin for a mastectomy. PMID- 11041045 TI - How to draw the ellipse of skin for a mastectomy. PMID- 11041046 TI - How to draw the ellipse of skin for a mastectomy. PMID- 11041047 TI - The management of women with breast symptoms referred to secondary care clinics in Sheffield: implications for improving local services. PMID- 11041048 TI - Solitary nodal metastasis presenting as branchial cysts: a diagnostic pitfall. PMID- 11041049 TI - Solitary nodal metastasis presenting as branchial cysts: a diagnostic pitfall. PMID- 11041050 TI - Solitary nodal metastasis presenting as branchial cysts: a diagnostic pitfall. PMID- 11041051 TI - Angiogenesis in cancer: the role of endothelin-1. PMID- 11041052 TI - A review of alternative approaches in the management of iatrogenic femoral pseudoaneurysms. PMID- 11041053 TI - Microorganisms in the aetiology of atherosclerosis. AB - Recent publications have suggested that infective pathogens might play an important role in the pathogenesis of atherosclerosis. This review focuses on these microorganisms in the process of atherosclerosis. The results of in vitro studies, animal studies, tissue studies, and serological studies will be summarised, followed by an overall conclusion concerning the strength of the association of the microorganism with the pathogenesis of atherosclerosis. The role of the bacteria Chlamydia pneumoniae and Helicobacter pylori, and the viruses human immunodeficiency virus, coxsackie B virus, cytomegalovirus, Epstein Barr virus, herpes simplex virus, and measles virus will be discussed. PMID- 11041054 TI - Morphological identification of the patterns of prostatic intraepithelial neoplasia and their importance. AB - High grade prostatic intraepithelial neoplasia (PIN) is the most likely precursor of prostatic carcinoma. PIN has a high predictive value as a marker for carcinoma, and its identification in biopsy specimens warrants repeat biopsy for concurrent or subsequent carcinoma. The only methods of detection are biopsy and transurethral resection; PIN does not greatly raise the concentration of serum prostate specific antigen (PSA) or its derivatives, does not induce a palpable mass, and cannot be detected by ultrasound. Androgen deprivation decreases the prevalence and extent of PIN, suggesting that this form of treatment might play a role in chemoprevention. Radiotherapy is also associated with a decreased incidence of PIN. PMID- 11041056 TI - Is it time to give up the crossmatch? PMID- 11041055 TI - Laser capture microdissection in pathology. AB - The molecular examination of pathologically altered cells and tissues at the DNA, RNA, and protein level has revolutionised research and diagnostics in pathology. However, the inherent heterogeneity of primary tissues with an admixture of various reactive cell populations can affect the outcome and interpretation of molecular studies. Recently, microdissection of tissue sections and cytological preparations has been used increasingly for the isolation of homogeneous, morphologically identified cell populations, thus overcoming the obstacle of tissue complexity. In conjunction with sensitive analytical techniques, such as the polymerase chain reaction, microdissection allows precise in vivo examination of cell populations, such as carcinoma in situ or the malignant cells of Hodgkin's disease, which are otherwise inaccessible for conventional molecular studies. However, most microdissection techniques are very time consuming and require a high degree of manual dexterity, which limits their practical use. Laser capture microdissection (LCM), a novel technique developed at the National Cancer Institute, is an important advance in terms of speed, ease of use, and versatility of microdissection. LCM is based on the adherence of visually selected cells to a thermoplastic membrane, which overlies the dehydrated tissue section and is focally melted by triggering of a low energy infrared laser pulse. The melted membrane forms a composite with the selected tissue area, which can be removed by simple lifting of the membrane. LCM can be applied to a wide range of cell and tissue preparations including paraffin wax embedded material. The use of immunohistochemical stains allows the selection of cells according to phenotypic and functional characteristics. Depending on the starting material, DNA, good quality mRNA, and proteins can be extracted successfully from captured tissue fragments, down to the single cell level. In combination with techniques like expression library construction, cDNA array hybridisation and differential display, LCM will allow the establishment of "genetic fingerprints" of specific pathological lesions, especially malignant neoplasms. In addition to the identification of new diagnostic and prognostic markers, this approach could help in establishing individualised treatments tailored to the molecular profile of a tumour. This review provides an overview of the technique of LCM, summarises current applications and new methodical approaches, and tries to give a perspective on future developments. In addition, LCM is compared with other recently developed laser microdissection techniques. PMID- 11041057 TI - Recent striking changes in histological differentiation and rate of human papillomavirus infection in squamous cell carcinoma of the lung in Okinawa, a subtropical island in southern Japan. AB - AIMS: The incidence of lung cancer in Okinawa has been the highest in Japan since 1975, and squamous cell carcinoma (SCC), especially the well differentiated form, is the most prevalent form in Okinawa, although well differentiated SCC is relatively rare in mainland Japan. Furthermore, a high proportion of SCC of the lung in Okinawa was positive for human papillomavirus (HPV). In this study, we report recent striking changes in histological features and in the incidence of HPV infection. METHODS: In Okinawa between 1986 and 1998, 1109 surgically resected lung tumours were examined histopathologically. In addition, human papillomavirus infection was detected by the polymerase chain reaction and Southern blot analysis in SCC cases reported in 1993 and 1995-8. Non-isotopic in situ hybridisation of HPV DNA was also carried out. RESULTS: Up until 1994 SCC, especially the well differentiated form, was the most prevalent type of tumour. However, since 1995 the number of such cases has diminished steadily, accompanied by a slight rise in the incidence of adenocarcinoma. Although most present and past patients are heavy smokers, the incidence of SCC, especially the well differentiated form, continues to decrease steadily. Furthermore, in 1993, HPV was detected in 79% of all cases, and was particularly prevalent in the well differentiated form, but the rate fell to 68% in 1995, 35% in 1996, 23% in 1997, and 24% in 1998. The age distribution of patients, the male to female ratio, and the number of tumours overexpressing p53 protein did not change significantly over the study period, and thus did not correlate with changes in the differentiation of SCC. CONCLUSIONS: The decreasing incidence of viral infection correlates strongly with the falling numbers of SCC cases, especially well differentiated cases. These findings suggest that HPV might be involved in the development of SCC of the lung, affecting the histological differentiation of SCC in particular, at least in Okinawa, a subtropical island in southern Japan. PMID- 11041058 TI - Histopathological detection of lymph node metastases from colorectal carcinoma. AB - AIM: To evaluate whether the assessment of multiple sections from retrieved nodes yields an increased number of metastases compared with the number that would be detected by the commonly applied method of microscopy of a single section of lymph node only. METHODS: A prospective study of 72 colorectal carcinoma resection specimens. Lymph node sampling was based on the current guidelines for the detection of breast cancer metastases in axillary nodes. Lymph nodes up to approximately 5 mm in maximum extent were processed in entirety, without prior sectioning, and assessed histologically at three levels; larger lymph nodes were processed in entirety as multiple sections and histologically assessed at one level. RESULTS: From a total of 72 carcinomas, eight were Dukes's A, 26 were Dukes's B, and 38 were Dukes's C. The mean and median numbers of nodes identified were 13 and 12, respectively (range, three to 44). Of the Dukes's C cases, four contained lymph node metastases identified by our method that might have gone undetected by the current, generally applied method. In one case, this led to the detection of the only nodal metastasis present and therefore "upstaged" the tumour from Dukes's B to C. On average, six extra tissue blocks were processed for each case in applying this method. CONCLUSION: The assessment of multiple sections of lymph nodes from colorectal specimens leads to the detection of only a small number of additional nodal metastases. The method involves increased workload for pathologists and laboratory staff. PMID- 11041059 TI - Frequency of oestrogen and progesterone receptor positivity by immunohistochemical analysis in 7016 breast carcinomas: correlation with patient age, assay sensitivity, threshold value, and mammographic screening. AB - AIMS: A routine immunohistochemical (IHC) assay is now commonly used for determining the oestrogen receptor (ER) and progesterone receptor (PR) status of women with breast cancer. To date, no large studies have been conducted that report the expected frequency of receptor positivity in relation to patient age and sensitivity of the IHC assay. Data on 7016 breast carcinomas from 71 laboratories were analysed to determine the frequency of receptor positivity and investigate possible causes of the observed variation in detection rates. METHODS: Members of UK NEQAS-ICC (UK National External Quality Assessment Scheme for Immunocytochemistry) provided data on the receptor status of cases routinely assayed in their departments over a period of two to 26 months between June 1996 and September 1998. Data on 7016 breast carcinomas were stratified according to patient age and receptor status. Frequency of receptor positivity was correlated with IHC assay sensitivity, the threshold value used to determine receptor positivity, and the presence or absence of mammographic screening in the hospitals or clinics served by the laboratories. RESULTS: The highest proportion of receptor positive cases occurred in patients in the age ranges > 65 years for ER and 41-50 years for PR. There was a significant positive correlation between frequency of receptor positivity and the sensitivity of the IHC assay, for both ER (rs = 0.346; p = 0.019; two tailed) and PR (rs = 0.561; p = 0.003; two tailed). The mean frequency of receptor positivity for laboratories using the same 10% threshold value was 77% for ER (95% confidence interval (CI), 74% to 80%) in laboratories with high sensitivity and 72% (95% CI, 68% to 76%) for those with low assay sensitivity (p = 0.025). For PR, the mean frequency of receptor positivity for laboratories using the same 10% threshold value and having high assay sensitivity was 63% (95% CI, 57% to 69%), and 51% (95% CI, 38% to 65%) for laboratories with assays of low sensitivity (p = 0.022). The mean frequency of ER positivity for laboratories serving hospitals and clinics where mammographic screening does and does not take place was 73.4% and 75.7%, respectively (p = 0.302; two tailed). CONCLUSIONS: Of the parameters investigated, patient age and IHC assay sensitivity were found to be the main variables influencing the frequency of receptor positivity. We recommend the range of receptor values obtained by laboratories achieving high assay sensitivity as a useful guide against which all laboratories can gauge their own results. PMID- 11041061 TI - Thymidine phosphorylase expression in normal and hyperplastic endometrium. AB - AIMS: To investigate the expression of thymidine phosphorylase (TP), a known angiogenic factor for endothelial cells, in normally cycling endometrium and various forms of endometrial hyperplasia. METHODS: TP expression was assessed with the P-GF.44C monoclonal antibody, using the alkaline phosphatase anti alkaline phosphatase method. Ninety two normal and hyperplastic endometria were studied. RESULTS: In normal proliferative endometrium, TP is found exclusively in the basal layer and the inner third of the functionalis; expression is cytoplasmic in glandular epithelium and nuclear in stromal cells. It is invariably patchy. This immunohistochemical picture remains almost unaltered during the early and mid secretory phase of the normal menstrual cycle but, most impressively, TP is expressed uniformly in the epithelium of all endometrial glands towards the end of the cycle. At this stage, expression is mixed nuclear/cytoplasmic and there is very little stromal nuclear staining. In simple endometrial hyperplasia, the staining pattern for TP is identical to normal proliferative endometrium, with a distribution that is usually limited to a few rather weakly proliferating glands and to the adjacent periglandular stroma of the deep endometrium. The distribution is more extensive in complex and atypical endometrial hyperplasias, where a mixed nuclear/cytoplasmic pattern usually prevails over the pure cytoplasmic reaction. CONCLUSIONS: TP is expressed consistently in normal and hyperplastic endometrium, suggesting a role in physiological and pathological angiogenesis. In normal endometrium, TP has a definite pattern of distribution, which is dependent on the phase of the menstrual cycle, whereas in all forms of endometrial hyperplasia the enzyme is randomly distributed and lacks an orderly pattern. PMID- 11041060 TI - Computerised morphometrical analysis in endometrial hyperplasia for the prediction of cancer development. A long-term retrospective study from northern Norway. AB - AIMS: To evaluate and compare the long term prognostic value of the WHO classification and the computerised multivariate morphometrical D score in endometrial hyperplasia. To test the reproducibility of the D score in two different centres. METHODS: Histopathological WHO classification and computerised morphometrical analysis using the D score (< 0, high risk; > 1, low risk; 0-1, uncertain) in a population based study from northern Norway of archival dilatation and curettage material from 68 women with 10-20 years of follow up. RESULTS: Of the 68 patients included in the study, 18 developed cancer. The sensitivity and specificity of the D score (< 0 v > 1) were 100% and 78%, respectively, which was better than the WHO classification (89% and 60%, respectively). The negative and positive predictive values for the D score were 100% and 58% and of the WHO classification 94% and 44%, respectively. This study found a slightly higher specificity for the D score than former retrospective studies, but otherwise the results were comparable. The D score results were reproducible between the two centres (R = 0.91; slope = 0.98; intercept = 0.3). CONCLUSIONS: D score assessment is a reproducible and more accurate predictor of outcome of endometrial hyperplasia than the WHO classification assessed by an experienced gynaecological pathologist. Routine application of the D score might reduce over and undertreatment of endometrial hyperplasia. PMID- 11041062 TI - The clinical spectrum of Clostridium sordellii bacteraemia: two case reports and a review of the literature. AB - Clostridium sordellii is rarely associated with disease in humans. Since its first report in 1922 only a few cases of bacteraemia have been reported. This report describes two cases of C sordellii bacteraemia; the oldest and youngest patients reported to date. The first, is a previously well 81 year old woman presented with perianal infection, which was later complicated by thrombosis of the aorta, and the second is a 12 year old boy with epilepsy who presented with an ear infection. These cases are also highlighted to demonstrate the wide spectrum of presentation of sordellii bacteraemia. PMID- 11041063 TI - Survival of leptospires in commercial blood culture systems revisited. AB - AIM: To assess the ability of commercial blood culture systems to maintain leptospires. METHODS: Nine different commercial blood culture bottles were compared for their ability to maintain four leptospiral strains at two temperatures, 30 degrees C and 37 degrees C. Bottles were subcultured at 48 hours, and one, two, three, and four week intervals and examined microscopically for the presence of viable leptospires. RESULTS: The results were comparable with those of an earlier study, which showed that different commercial blood culture systems varied in their ability to maintain leptospires. CONCLUSIONS: No single factor appears to influence the viability of leptospires in blood culture systems. In general, the combination of an aerobic blood culture and an incubation temperature of 30 degrees C enhances the viability of leptospires, and hence would increase the chances of their subsequent isolation from suspected cases of leptospiraemia. PMID- 11041064 TI - Evaluating the usefulness of the ICT tuberculosis test kit for the diagnosis of tuberculosis. AB - BACKGROUND: Early diagnosis of tuberculosis is crucial, especially in Korea, where tuberculosis is endemic. AIMS: To evaluate the validity of the ICT tuberculosis test (ICT) in early diagnosis of tuberculosis. METHODS: Sixty eight patients with tuberculosis were tested; 37 had no history of previous tuberculosis (patient group 1), and 31 had reactivated tuberculosis (patient group 2). The control groups comprised 77 subjects: 25 healthy adults, 35 hospital workers, and 17 inpatients with non-tuberculous respiratory diseases. RESULTS: The diagnostic sensitivities of ICT were 73% in patient group 1 and 87.1% in patient group 2. In two patients with extrapulmonary tuberculosis, both tested positive using ICT. The specificities of ICT were 88%, 94%, and 94% in healthy adults, hospital workers, and non-tuberculous patients, respectively. CONCLUSIONS: ICT is a useful tool for the diagnosis of tuberculosis. PMID- 11041065 TI - Demand management in urine cytology: a single cytospin slide is sufficient. AB - AIMS: Current practice in most laboratories stipulates the preparation of duplicate slides for the analysis of urine cytology specimens. This study evaluates whether the duplicate slide is necessary. METHODS: Cytospin diagnosis was assessed in three ways. First, all urine cytology preparations from a single month in 1998 were reviewed; the two slides for each case were reported separately and then the two reports on each case were examined for disparity. Second, the slides from all urine cytospin cases indexed as "suspicious" or "malignant" in 1998 were reviewed similarly. Third, 48 cytospin slides from 24 cases were divided into two randomised groups, which were reported and the two reports compared. Finally, the frequency of repeat specimen collection in cases that were deemed inadequate for diagnosis was also assessed. RESULTS: The cases from a single month (n = 129) were representative of the annual workload and showed no discrepancies of the type: suspicious or malignant/other. Of the 60 suspicious or malignant cases from 1998, there was no disparity in 50. The 10 cases with disparity were all suspicious on one slide and degenerate on the other. In the 24 randomised cases, there was no disparity in 21. The remainder were reported as suspicious or malignant/inadequate (that is, degenerate or acellular). After a report of inadequate for diagnosis, repeat samples were received in only 15% of cases. CONCLUSIONS: Using a single cytospin preparation causes minimal loss of clinically relevant information, but saves substantial resources (approximately 40%/case). A diagnosis of inadequate should prompt the collection of a repeat sample if the service is being used sensibly. PMID- 11041066 TI - Small intestinal infarction: a fatal complication of systemic oxalosis. AB - Primary hyperoxaluria is a rare genetic disorder characterised by calcium oxalate nephrolithiasis and nephrocalcinosis leading to renal failure, often with extra renal oxalate deposition (systemic oxalosis). Although ischaemic complications of crystal deposition in vessel walls are well recognised clinically, these usually take the form of peripheral limb or cutaneous ischaemia. This paper documents the first reported case of fatal intestinal infarction in a 49 year old woman with systemic oxalosis and advocates its consideration in the differential diagnosis of an acute abdomen in such patients. PMID- 11041067 TI - Rationalised virological electron microscope specimen testing policy. PMID- 11041068 TI - Aberrant CD10 expression by NHL. PMID- 11041069 TI - Demographic and nutritional trends among the elderly in developed and developing regions. AB - Aging for an individual and aging for a population are related but not the same. For an individual, aging first involves survival to more advanced years, which will inevitably be accompanied by progressive changes in the structure and function of somatic tissues due to a programmed failure of the organism maximally to invest in their maintenance. For a population, aging means an increase in the median age, a dual function of longer survival of individuals and a decrease in fertility. In the wilds of nature, and for all but the recent decades for the human species, survival beyond the peak reproductive years is rare. The hostility of natural selection has its greatest impact on the young and the old. Only captive fowl and beasts, domesticated animal species and Homo sapiens achieve long survival. For humans, life expectancy from birth and from any age thereafter is increasing; median ages of populations and the percentage of persons over 60 y are rising. The affluent developed countries led the way, but developing countries are closing the gap. As of about 1966, a majority of the world's elderly live in developing societies. The rarity of growing old left a dearth of knowledge in the domain of gerontological nutrition, both for lack of motivation to learn and lack of individuals and populations to study. The convening of this workshop signifies that the polarity of interest has shifted 180 degrees. Social, economic, physiological and psychological changes with aging and growing older can both be influenced by diet and influence eating patterns and nutritional status. Many assumptions have been made about these changes, but only recently, with concerted metabolic studies of nutrient requirements in healthy elderly and carefully designed multi-center surveys of the health and nutrition of older segments of populations can a true portrait of the issues be delineated. PMID- 11041070 TI - Effects of reduced energy intake on the biology of aging: the primate model. AB - Dietary energy restriction is the only proven method for extending lifespan and slowing aging in mammals, while maintaining health and vitality. Although the first experiments in this area were conducted over 60 y ago in rodents, possible applicability to primates has only been examined in controlled studies since 1987. Our project at the National Institute on Aging began with 3-0 male rhesus and 30 male squirrel monkeys of various ages over their respective life spans. Subsequently, it has been expanded to include female rhesus monkeys, and several other laboratories have initiated related studies. Experimental animals are generally fed 30% less than controls, and diets are supplemented with micronutrients to achieve undernutrition without malnutrition. These calorically restricted (CR) monkeys are lighter, with less fat and lean mass than controls. Bone mass is also slightly reduced, but in approximate proportion to the smaller body size. CR animals mature more slowly and achieve shorter stature than controls as well. Metabolically, CR monkeys have slightly lower body temperature and initial energy expenditure following onset of restriction, and better glucose tolerance and insulin sensitivity. The latter suggest a reduced predisposition towards diabetes as the animals age. Other potential anti-disease effects include biomarkers suggestive of lessened risk of cardiovascular disease and possibly cancer. Candidate biomarkers of aging, including the age-related decrease in plasma dehydroepiandrosterone sulfate (DHEAS), suggest that the CR animals may be aging more slowly than controls in some respects, although sufficient survival data will require more time to accumulate. In summary, nearly all CR effects detected in rodents, which have thus far been examined in primates, exhibit similar phenomenology. Potential applicability of these beneficial effects to humans is discussed. PMID- 11041071 TI - Physiology of aging with respect to gastrointestinal, circulatory and immune system changes and their significance for energy and protein metabolism. AB - Age-related changes in body composition can be considered the consequence of changes in energy and protein metabolism, while also having a leverage effect on protein and energy requirements. Changes in organ and systems weights obviously affect energy balance regulation. Considered at the system level, age-related changes are numerous, but it is still debated whether they are related to aging per se or to conditions (such as poor nutrition, disease, drug treatments etc.) that prevail in elderly persons. It is likely that most changes occuring in the gastrointestinal, circulatory and immune system do not affect energy and protein requirements at rest. However, aging is associated with difficulties in adapting to new environmental conditions that lead to stress. Repeated episodes of stress might lead to accumulation of deficits that can affect energy and protein balances. PMID- 11041072 TI - Anthropometry and methods of body composition measurement for research and field application in the elderly. AB - Evaluation of body composition is important in the study of human energy and protein metabolism as methods are available for quantifying energy stores and protein content at a single point in time; energy-protein balance can be monitored over time; and dynamic measures of energy and protein metabolism can be referenced to body mass and related measurable components for between-individual comparisons. This review emphasizes the need for considering subject age when developing body composition component prediction models that are applied in elderly populations. An overview of body composition research is provided that emphasizes compartment and level definitions and interrelations. Two broad method categories, mechanistic and descriptive, are then critically examined in relation to their role in energy-protein metabolism and aging research. Our collective review indicates that all major body composition components are now measurable using one or more methods that are based on non age-dependent assumptions. We also found that some methods, particularly descriptive field methods (eg anthropometry), may be based on age-sensitive assumptions and measurements and suggestions for future development of these methods are provided. Lastly, as body composition differences between races, cultures, and countries are now recognized, it would be useful to create international cooperative groups with the aim of developing widely applicable descriptive field methods based on simple available techniques such as anthropometry and bioimpedance analysis. PMID- 11041073 TI - Body weight and weight change and their health implications for the elderly. AB - After the age of 60 y, body weight on average tends to decrease. The contribution of fat mass to this weight loss is relatively small, but fat tends to be redistributed with advancing age toward more abdominal (particularly visceral) fat. Anthropometric data are relatively poor indicators of these aging processes. This may be one of the explanations why the relationship between high body mass index and mortality is less pronounced in older than in younger people. Reduced lipolysis in the visceral fat depot with aging is among potential explanations why increased visceral fat seems to be less harmful in elderly subjects compared to young adults. Even though the relative contribution of increased fat mass to mortality may be less pronounced in elderly people, the impact on disability and functional limitations is found to be important from both a clinical and a public health point of view. At the other end of the scale studies have shown that low body mass index and weight loss in the elderly are both strong predictors of subsequent mortality. This cannot be explained by effects of smoking and early mortality after baseline. There are only few systematic studies comparing the predictive validity of different anthropometric data for mortality. One recent prospective study showed that a high waist circumference (in nonsmoking men) may be a better predictor of all-cause mortality than high body mass index and waist/hip ratio. Low BMI was a better predictor of mortality than low waist circumference. In conclusion changes in body composition and fat distribution with aging are poorly captured by standard anthropometric data. Low lean body mass is probably better reflected by low BMI, whereas increased (abdominal) fatness is better reflected by increased waist circumference. PMID- 11041074 TI - Sarcopenia and its implications for the elderly. AB - Sarcopenia is the loss of muscle mass and strength with age. Sarcopenia is a part of normal aging, and occurs even in master athletes, although it is clearly accelerated by physical inactivity. Sarcopenia contributes to disability, reduced ability to cope with the stress of a major illness, and to mortality in the elderly. The etiology of sarcopenia is unclear, but several important factors have been identified. These include loss of alpha motor neurons, decline in muscle cell contractility, and several potential humoral factors, such as androgen and estrogen withdrawal and increase in production of catabolic cytokines. Treatment of sarcopenia with progressive resistance training is safe and effective, but dissemination of this technique to the general population has yet to occur. As the number of elderly persons increases exponentially in the new century, a public health approach to prevention and treatment of sarcopenia, based on increasing physical activity at all ages, will be crucial to avoiding an epidemic of disability in the future. PMID- 11041075 TI - Fat and protein redistribution with aging: metabolic considerations. AB - Aging is associated with a redistribution of both fat and lean tissue within the body. Intra-abdominal fat (IAF) accumulates more rapidly than total fat while the loss of lean body mass is mostly due to sarcopenia. Increase of visceral fat plays a major role in the pathogenesis of insulin resistance, which leads to type II diabetes and also to cardiovascular diseases. This review is focussed on the relationships that exist between the accumulation of IAF and insulin resistance during aging. The various methods available for assessing IAF are briefly reviewed; imaging techniques are the only reference methods, and their availability is limited. Insulin resistance that appears with aging is caused by accumulation of IAF, rather than by aging per se. Studies done in type II diabetic patients suggest that the metabolic link between increased IAF and insulin resistance could well be the increased availability and/or oxidation of free fatty acids. Physical inactivity certainly enhances both IAF accumulation and, more directly, insulin resistance. Independent and significant effects of menopause or of sarcopenia on insulin resistance remain to be established. The influence of hormonal changes, reduced fatty acid utilization, and resistance to leptin on IAF accumulation are also discussed. Although it is difficult to determine the independent influence of each of these factors, IAF accumulation seems to be a central and important determinant of cardiovascular risk. The last part of this review is devoted to protein metabolism and focused on the preservation of protein metabolism in the liver during aging. PMID- 11041076 TI - Taste and smell perception affect appetite and immunity in the elderly. AB - The losses in taste and smell that occur with advancing age can lead to poor appetite, inappropriate food choices, as well as decreased energy consumption. Decreased energy consumption can be associated with impaired protein and micronutrient status and may induce subclinical deficiencies that directly impact function. Most nutritional interventions in the elderly do not compensate for taste and smell losses and complaints. For example, cancer is a medical condition in which conventional nutritional interventions (that do not compensate for taste and smell losses) are ineffective. Evidence is now emerging that suggests compensation for taste and smell losses with flavor-enhanced food can improve palatability and/or intake, increase salivary flow and immunity, reduce chemosensory complaints in both healthy and sick elderly, and lessen the need for table salt. PMID- 11041077 TI - Regulation of energy intake in relation to metabolic state and nutritional status. AB - Inadequate energy intake can be an important contributor to weight loss in older individuals. This review highlights recent studies on possible causes of negative energy balance in older individuals. Studies of the regulation of food intake suggest that aging is associated with a significant impairment in the regulation of food intake that inhibits appropriate short-term and long-term compensation for imposed alterations in energy intake. The combination of a reduced ability to regulate energy intake, decreased sensory-specific satiety, and disadvantageous social factors such as functional limitations, social isolation and depression, increases the risk of negative energy balance leading to weight loss in older individuals. PMID- 11041078 TI - Determinants of macronutrient intake in elderly people. AB - An understanding of the determinants of nutritional intake may be helpful in the early detection and prevention of malnutrition. A vast amount of literature exists on factors influencing daily energy or food intake. Although far less is known about population and subject characteristics influencing the macronutrient composition of the diet, associations appear to exist with cultural factors, snacking and disease. The literature does not provide sufficient evidence in support of relationships with other factors such as living situation, dentition, disability, depression and drug use, partly because information on determinants of macronutrient intake is scarce. PMID- 11041079 TI - Mechanisms of changes in basal metabolism during ageing. AB - A considerable number of physiological functions are known to show a gradual decline with increasing age. However, the effects of ageing differ widely between organ systems. It is believed that basal metabolic rate (BMR) falls dramatically with age. These observations, largely based on cross-sectional surveys, are discussed in light of our present understanding of the biology of ageing. This paper reviews both the longitudinal and cross-sectional studies of BMR and presents evidence that the fall in BMR with ageing may be less dramatic than previously perceived. Indeed, some subjects may show an increase in BMR with ageing. The mechanism of changes in BMR during ageing will be discussed. Organ weight changes appear to have a profound impact on BMR. The use of BMR to predict total energy expenditure in the 'old elderly' (>75 y) is unlikely to be of any practical use due to wide intra- and inter-individual variation in BMR. This wide intra- and inter-individual variation in BMR is due to illness, disease and other metabolic disorders seen in the elderly. Finally, the importance of measuring BMR in elderly populations for its use in clinical medicine will be discussed. PMID- 11041080 TI - Total energy expenditure in the elderly. AB - Chronic diseases and disabilities increase with age, affecting more than 60% of those over 75 y, and limiting activities in about half of them. Therefore, total energy expenditure (TEE) and its components are assessed separately in health and disease. An analysis of 568 doubly labelled water measurements in 'healthy' subjects (184 measurements in subjects over 65 years) suggests that there is a decrease of 0.69 and 0.43 MJ/day/decade respectively in men (standard weight 75 kg) and women (standard weight 67 kg). Physical activity (PA) accounted for 46% of the decrease in TEE, basal metabolic rate (BMR) for 44% of the decrease and thermogenesis (T) for the remaining 10%. TEE was found to be 10.79+/-2.09 and 8.62+/-1.49 MJ/day in 150 men and 100 women aged over 60 y, respectively. Of the total variance in TEE, measured with doubly labelled water over a 2 week period, 69% was considered to be due to differences between individuals, and 31% to differences within individuals. The variance due to PA plus T was threefold greater than that due to BMR. Physiological factors were far more important than methodological factors in influencing measurements of TEE, BMR and PA+T. An analysis of 136 measurements of TEE (doubly labelled water and bicarbonate-urea methods) in free-living elderly patients suffering from a variety of diseases suggests a frequent decrease in TEE, which may occur despite an increase in BMR. This is largely due to a reduction is PA (eg up to approximately 50% reduction), but in some cases it is also due to a reduction in BMR (loss of body weight). More comprehensive information is required about TEE and its components, partly because of a probable selection bias in recruitment of subjects participating in specific tracer studies, and partly because of the variable effects of different diseases and factors that operate at different times in the course of the same disease. PMID- 11041081 TI - Assessment of energy requirements in elderly populations. AB - The daily energy requirements of older adults are unclear. Aging results in a decline in daily energy expenditure and intake, which are associated with greater gains in body fatness and a subsequently higher risk of morbidity and mortality. Understanding the energy requirements of older populations, therefore, has important clinical implications. Current world-wide recommendations suggest that energy needs of individuals above 50 y are of 1.51 times resting energy expenditure. Limited data suggest that this may be an underestimation of energy needs in older Caucasian women and men. In contrast, current energy requirement recommendations may be appropriate for older African-American women, because of their low rate of daily energy expenditure for their metabolic size. Aging is also associated with Alzheimer's and Parkinson's and recent data suggest that individuals with these chronic diseases actually have energy requirements that are at or below current recommendations. Physical activity is the most modifiable and variable component of daily energy expenditure and, therefore, energy requirements. Current data suggest that an inexpensive and simple measurement of physical activity is difficult in older adults, which limits our ability to accurately determine energy needs. Overall, current data suggest that energy needs of older adults may be higher than current world-wide recommendations, although future prospective data are needed in healthy, ethnic, and diseased populations. Accurate, unobtrusive, and inexpensive methods to measure physical activity are also needed to assess energy requirements. PMID- 11041082 TI - Carbohydrate metabolism in the elderly. AB - In this short review we summarize the effect of age on glucose homeostasis. The concept of decreased glucose tolerance with increasing age is introduced, followed by evidence for this phenomenon. Specifically we review the evidence for changes in fasting glucose as a function of age and the effect of age on HbA1c. The role of age on hepatic glucose production and glucose uptake is then discussed in detail and we review the evidence that supports the concept that with advancing age hepatic glucose sensitivity to insulin is unaltered. We then review the large evidence for the role of age on the purported decrease in peripheral tissue sensitivity to insulin and conclude that the issue is unsettled. The decrease attributed to age is no longer significant when confounders are controlled for, the largest being obesity. We next present evidence that beta-cell sensitivity to glucose remains intact with aging. A review of age-related disorders due to hyperglycemia and confounding effects on the relationships of age and glucose tolerance is presented next. Finally we present new evidence that when the revised criteria for the diagnosis of type 2 diabetics as proposed by the American Diabetes Association and WHO are used, a greater percentage of the elderly will not be diagnosed. We conclude that, although glucose intolerance increases with aging, which is accompanied with other disorders, it is possible to ameliorate this effect with alteration of diet and exercise. PMID- 11041083 TI - Lipid metabolism in the elderly. AB - Adiposity increases with age. The size of the adipose tissue mass is determined by the balance between the recruitment of lipid substrates (ie free fatty acids) from adipose tissue and their subsequent oxidation by respiring tissues. Thus, change in the liberation of free fatty acids from adipocytes, the capacity of respiring tissue to oxidize free fatty acids or a combination of both may contribute to the age-related increase in body fat. This review focuses on studies that have examined the effect of age on free fatty acid release and the capacity of respiring tissues to oxidize fat. In vitro studies have shown that hormonal and pharmacological stimulation of lipolysis diminished with age. Despite this cellular defect, however, in vivo studies suggest that fatty acids are recruited from adipose tissue in excess of the energy demands of the body in older individuals. The capacity of respiring tissues, in particular skeletal muscle, to oxidize fat declines with age. The age-related decrease in fat oxidation is related to a reduction in both the quantity and oxidative capacity of respiring tissue. Taken together, these results suggest that an age-related decrease in the capacity of respiring tissues to oxidize fat, rather than decreased free fatty acid release, is a more likely determinant of lipid imbalance and the age-related increase in adiposity. Interventions designed to increase the mass or oxidative capacity of respiring tissue, therefore, may be effective in counteracting the age-related reduction in fat oxidation. PMID- 11041084 TI - Role of insulin in age-related changes in macronutrient metabolism. AB - Age is associated with an increase in body fat mass and a decrease of protein mass. As body substrate turnover is under insulin control, defects in insulin secretion and/or action may in part account for these changes. As regards secretion, current evidence suggest that no clear defect in insulin secretion is found in the aged. The wide spectrum of glucose tolerance of the elderly may be associated with different patterns of insulin secretion. Insulin sensitivity to glucose metabolism is more or less normal in the aged, despite subtle delays in the onset of its action. Normalization of the data by either body weight or lean body mass is important in defining the insulin sensitivity of the elderly. Increased rates of free fatty acid (FFA) flux and oxidation rates have been found in healthy elderly subjects, both when post-absorptive and during hyperinsulinemia. These differences however disappeared following normalization by fat mass, suggesting that FFA kinetics reflect the established changes in fat mass. Thus, the mechanism(s) leading to an increase in the fat mass in elderly cannot simply be derived from studies of fat kinetics. The operation of the Randle cycle (ie, inverse relationships between fat and glucose oxidation) in the elderly has also been suggested. Finally, the insulin effects on whole-body amino acid and protein metabolism do not seem to be impaired in the aged. However, in the human muscle a decreased synthesis of contractile as well as mitochondrial proteins was found, in association with decreased specific gene expression. The degree of physical activity probably interacts with these changes, possibly playing a causative role. The possible interaction between insulin and exercise in the maintenance of muscle mass in the elderly needs to be studied further. PMID- 11041085 TI - Protein and amino acid requirements in the elderly. AB - Estimates of protein and amino acid requirements in this paper are proposed for healthy elderly people. The estimate of protein requirement was based on nitrogen (N) balance, as well as functional indicators such as immune function or muscle strength. Data suggest that the protein requirement for nitrogen equilibrium in the elderly, is greater than 0.8 gm/kg body weight/day. There do not appear to be any adverse consequences with protein intakes that are about 1 gm/kg body weight/day. The tentative recommendation in this paper is higher than the current mean recommended intake of protein (FAO/WHO/UNU, 1985). However, because of methodological difficulties, the data does not allow for a confident prediction of what the exact level of protein intake should be. Further studies are needed to come to a firm conclusion on the exact protein requirement. Indispensable amino acid requirements based on nitrogen balance data, in the elderly, are fragmentary and conflicting. These requirements can alternatively be based on obligatory nitrogen loss, for which data is available in the elderly. The overlap of the obligatory nitrogen loss between the young and the elderly, suggest that the amino acid requirement based on this technique is similar in young and elderly individuals. Tracer based techniques measuring amino acid balance at different amino acid intakes, also support the view that there are no differences in the amino acid requirements between young and elderly people. In general, these amino acid requirement studies have been performed in healthy USA subjects, and data is needed to know if these estimates can be extended to populations from other, less-developed countries. PMID- 11041086 TI - Relationships among diet, physical activity and other lifestyle factors and debilitating diseases in the elderly. AB - Diet and physical activity are two major lifestyle factors that play a role in the prevention or management of debilitating conditions affecting older people. Both under- and overnutrition predispose to diseases. Low sodium and high potassium intakes, as well as the consumption of fruits and vegetables are associated with a reduction of hypertension and diseases arising from hypertension such as stroke and dementia. Dietary patterns (consumption of quantity and types of fats, cholesterol, vegetable oils, fish) are important in the prevention of coronary heart disease. Calcium and vitamin D intakes are important factors in the development of osteoporosis, while various dietary factors have been linked to the development of cancer. Physical activity is important in the prevention of functional decline and increased survival, reduced incidence of falls and fractures, and has various cardiovascular health benefits. Apart from prevention of diseases, exercise also has an important role in improving function in some chronic diseases such as heart failure or chronic obstructive pulmonary disease. Both diet and exercise interact, so that public health recommendations often take the form of lifestyle modification advice in the prevention of disease and disability. PMID- 11041087 TI - Interventions aimed at dietary and lifestyle changes to promote healthy aging. AB - Desirable dietary habits and other lifestyle practices reduce premature mortality and compress the period of morbidity experienced towards the end of life. Aging adults are at risk of nutritionally inadequate diets especially in relation to protein, vitamins D, B1, B6, B12, fluid and other food components. Interventions aimed at ensuring dietary adequacy also need to consider the social and cultural aspects of eating as food is fundamental to a person's well-being and quality of life. The nutrition-related health problems associated with aging such as frailty, depression, incontinence and chronic non-communicable diseases should be identified in both the individual and in the community before dietary and other health interventions are implemented. In older adults, these dietary and health promoting interventions should then focus on maximizing function and quality of life, be acceptable and finally, measurable in terms of effectiveness. PMID- 11041088 TI - Report of the IDECG Working Group on the biology of aging. The International Dietary Energy Consultative Group. PMID- 11041089 TI - Report of the IDECG Working Group on body weight and body composition of the elderly. PMID- 11041090 TI - Report of the IDECG Working Group on energy and macronutrient metabolism and requirements of the elderly. PMID- 11041091 TI - Report of the IDECG Working Group on the role of lifestyle including nutrition for the health of the elderly. PMID- 11041092 TI - Evaluation of the heart with magnetic resonance imaging. AB - After compensating for two kinds of motion artifacts caused by cardiac beating and respiration, cardiac magnetic resonance (MR) imaging is now feasible for the diagnosis of various cardiac diseases. Taking cost-effectiveness into consideration, this paper reviews the experiences of preferable indications of cardiac MR imaging by demonstrating the characteristic preciseness and uniqueness that play an important role in obtaining time-volume curves consisting of the theoretically most accurate measurements of left and right ventricular volumes, in overall evaluation of the left ventricular apex and the right ventricle, in delineating the wide range of the coronary arterial tree, in measuring the most precise blood flow volume through the cross-sectional images of the vessels, and in assessing the spatial derivative of the blood flow velocity at the vessel wall, i.e., wall shear rate. PMID- 11041093 TI - Leukocyte-depleted continuous blood cardioplegia for coronary artery bypass grafting. AB - Many cardiac surgeries are performed with blood cardioplegia. However, some studies suggest that activated neutrophils form blood cardioplegia can cause reperfusion injury. In this study we assessed myocardial protection using a leukocyte-depleted cardioplegic solution. Patients undergoing elective coronary artery bypass grafting (CABG) with continuous blood cardioplegia were divided into two groups: the LD group, which received leukocyte-depleted blood cardioplegia (n = 11); and the control group, which received nonfiltered blood cardioplegia (n = 11). IL-6, IL-8, CK-MB, and troponin T were measured in the coronary sinus blood immediately after the release of the aortic cross-clamp. Cytokine concentrations were also measured upon the patient's return to the ICU. The total dopamine and dobutamine doses, hemodynamic measurements after surgery, and the leukocyte filtration rate were also measured. During antegrade cardioplegia infusion, leukocytes were almost completely removed (filtration rate: 85.8+/-4.0%). However, during terminal warm cardioplegia, leukocyte removal decreased (filtration rate: 39.9+/-7.8%). Immediately after the release of the aortic cross-clamp, plasma CK-MB and troponin T concentrations were significantly lower in the LD group (17.7+/-1.9 U/l and 0.017+/-0.002 ng/ml, respectively) than in the control group (30.3+/-3.6 U/l and 0.072+/-0.029 ng/ml, respectively). The IL-6 and IL-8 concentrations were similar in the LD group and the control group. After the return to the ICU, the CK-MB and troponin T concentrations were similar in the two groups. No significant differences were found in the total doses of dopamine or dobutamine after surgery in the two groups (99+/-77 vs 101+/-128 microg/kg/min). No significant differences were found in the hemodynamic parameters after surgery in the two groups. In patients undergoing CABG with continuous blood cardioplegia, leukocyte-depleted blood cardioplegic solution may attenuate reperfusion injury. PMID- 11041094 TI - Dobutamine stress echocardiography for the diagnosis of myocardial viability: assessment of left ventricular systolic velocities in longitudinal axis by pulsed Doppler tissue imaging. AB - Dobutamine (DOB) stress two-dimensional echocardiography is an established method for the detection of viable myocardium, but conventional assessment of wall motion is subjective. We measured quantitatively the left ventricular systolic velocities along the longitudinal axis by pulsed Doppler tissue imaging (DTI). In 30 patients with previous myocardial infarction, pulsed DTI focused on the infarct area was performed from an apical two- or four-chamber view before and during DOB (10 microg/kg/min) stress one day before coronary angioplasty. We calculated peak systolic velocity (S), regional pre-ejection period (PEP, the time interval from the onset of QRS to the onset of systolic wave) and regional ejection time (ET). Left ventriculography was obtained before and 3 months after coronary angioplasty to assess regional wall motion. Improvement of abnormal wall motion was observed in 19 patients (group P) but not in 11 (group N). Group P had significantly larger S and smaller PEP/ET than group N during DOB stress, although there were no significant differences in these indices between the groups at baseline. As a consequence, group P had a significantly larger percent change in S and a smaller percent change in PEP/ET than group N (164+/-39 vs 117+/-20% and 88+/-17 vs 116+/-29%, respectively, p < 0.01). It is suggested that the quantitative measurement of longitudinal systolic velocities during DOB stress by DTI is useful for the precise assessment of myocardial viability. PMID- 11041095 TI - What determines the heart rate response to exercise in patients with atrial fibrillation? AB - We evaluated the factors that determine the heart rate response to exercise in 60 patients with atrial fibrillation (25 men and 35 women, with a mean age of 61+/ 10 years) who underwent symptom limited cardiopulmonary exercise testing with blood sampling of atrial natriuretic peptide (ANP), 2-dimensional echocardiography and cardiac catheterization. Atrial muscles resected during the Maze operation were examined histologically in 12 patients. The heart rate response to exercise depended on the severity of the atrial organic injury, which was expressed as left atrial diameter, ANP secretion during the maximal exercise testing and the histological findings of atrial tissue. Conversely, we believe that the severity of the atrial injury can be predicted from the heart rate response to exercise in patients with atrial fibrillation. PMID- 11041096 TI - Right atrial appendage function in patients with chronic nonvalvular atrial fibrillation. AB - To assess right atrial appendage (RAA) flow and its possible relationship to left atrial appendage (LAA) flow in chronic nonvalvular atrial fibrillation (AF), transesophageal echocardiography (TEE) was performed in 26 patients with chronic nonvalvular AF (group I). For the purpose of comparison, an additional group of 27 patients with chronic valvular AF due to mitral stenosis (group II) was analyzed. The clinically estimated duration of AF in group I was significantly longer than that of group II (8.7+/-3.4 versus 2.7+/-1.1 years). Although right atrial size and RAA maximal area were larger in group I than those in group II, left atrial size was larger in group II than that in group I. Group II had larger LAA maximal areas than group I, but this difference did not reach statistical significance. The two groups were not different with respect to the RAA or LAA emptying velocities. Significant correlations were observed between echocardiographic parameters of the two atria in patients with nonvalvular AF (r range, 0.4 to 0.7). In contrast, in patients with valvular AF, no correlation was observed between the echocardiographic parameters of the two atria (appendage emptying velocity, r = 0.38, p = 0.051; atrial size, r = -0.03, p = 0.89; maximal appendage area, r = 0.07, p = 0.75, respectively). There were no significant differences in the presence of right and left atrial spontaneous echo contrast and thrombus between the groups. All of the right and left atrial thrombi were confined to their respective appendages and were found in the atria with spontaneous echo contrast. Both RAA and LAA thrombi were present in one patient. In conclusion, our findings suggest that AF could affect both atria equally in nonvalvular AF, in contrast to valvular AF. Therefore, the assessment of RAA function as well as LAA may be important in patients with chronic nonvalvular AF. PMID- 11041097 TI - Prolongation of QT interval and ventricular septal hypertrophy. AB - Long QT syndrome (LQTS) is a prime example of interplay between molecular biology, cellular physiology, and organ physiology. Both the congenital and acquired forms of LQTS are due to intrinsic and/or acquired abnormalities of the ionic currents responsible for cardiac repolarization. We analyzed the QTc interval, QRS axes and interventricular septal thickness (IVST) in 41 patients who had a prolonged QT interval on routine electrocardiography (ECG) (5 females and 36 males, mean age 65+/-13 years). The QRS axis of patients in the LQTS group (27+/-49 degrees) was significantly lower (p < 0.05) than in the control group (46+/-26 degrees). However, the IVST in the LQTS group (10+/-2 mm) was significantly thicker than in the control group (9+/-1 mm) (p < 0.05), while the WTd was not significantly different. The QTc interval in patients with ventricular septal hypertrophy (IVST > or = 12 min, 478.8+/-7.9 msec) was significantly longer (p < 0.05) than in the normal group (IVST < 12 mm, 472.1+/ 17.5 msec). In conclusion, the results of this preliminary study suggest that prolongation of the QT interval on ECG should prompt screening for electrocardiographic evidence of ventricular hypertrophy in patients with this disease. PMID- 11041098 TI - Transcatheter closure of atrial septal defect with a CardioSEAL device. AB - Transcatheter closure of an interatrial septal defect (ASD) with a CardioSEAL device was successfully performed on 12 Taiwanese children. There were five boys and seven girls, aged from 3.6-13.9 (8.3+/-3.2) years and with body weight of 15 57 (33.7+/-14.7) kgs. After one year of follow-up studies, which included physical examination, ECG, chest X-ray and echocardiography, complete closure of ASD was achieved in nine (75%) patients. Two children with a trivial residual shunt were asymptomatic and without audible cardiac murmur. A girl had a small residual left to right atrial shunt by color Doppler echocardiography, but without audible cardiac murmur. There were no immediate or intermediate complications. Transcatheter implantation of the CardioSEAL device is a safe and proper treatment for children with non-complicated small to medium secundum ASD. PMID- 11041099 TI - Preceding stimulus frequency-dependent potentiation of the postrest shortening of the action potential duration in rabbits. AB - Action potential duration (APD) in rabbit ventricular myocardium shortens after a rest period (postrest shortening). However, the effects of preceding stimulus frequency on the postrest shortening have not been elucidated. We recorded transmembrane action potentials (TAPs) and monophasic action potentials (MAPs) from the rabbit ventricle. In in vitro experiments. repetitive regular stimuli (S1) at cycle lengths ranging between 500 to 3000 ms were followed by a single extrastimulus (S2) at a coupling interval of 5000 ms. A decrease in S1S1 interval resulted in a progressive shortening of the duration of TAP (TAPD) elicited by S2 (S2-TAPD), which was potentiated by increasing extracellular calcium concentration ([Ca2+]o) or application of ouabain and was inhibited by lowering [Ca2+]o or verapamil. Application of ryanodine was most effective in lengthening S2-TAPD following a short S1S1 interval. 4-aminopyridine and E4031 caused marked lengthening of S2-TAPD when S1S1 was long. However, the lengthening effect was attenuated and disappeared with a shorter S1S1 interval. In in vivo experiments, regular ventricular pacing (S1) at cycle lengths ranging between 250 to 1000 ms was followed by a single extrastimulus (S2) with a coupling interval (S1S2) of 1500 ms. A decrease in the S1S1 interval also resulted in progressive shortening of the duration of MAP elicited by S2. Our results indicate that the postrest shortening is potentiated by an increase in the preceding stimulus frequency in the rabbit ventricle, in which the function of sarcoplasmic reticulum may play a significant role. PMID- 11041100 TI - Inducible nitric oxide-mediated myocardial apoptosis contributes to graft failure during acute cardiac allograft rejection in mice. AB - The mechanism through which nitric oxide (NO) mediates cardiac myocyte death during acute cardiac rejection has not been fully delineated. We sought to determine whether NO promotes myocardial apoptosis and contributes to graft failure during acute cardiac rejection in a murine model. Heterotopic cardiac transplantation was performed from Balb/c (H-2d) to C3H/He mice (H-2k). Recipients were treated with aminoguanidine (AG) at 400 mg/kg every day after surgery. As references, we used isografts in Balb/c mice with and without AG treatment (400 mg/kg/day). Graft survival, histological changes and serum NO levels were assessed. Intra-graft apoptosis was evaluated using a DNA fragmentation detection assay (TUNEL method) and DNA laddering. Significant prolongation of graft survival was observed in allografts treated with AG in comparison with nontreated allografts. Serum NO levels, which peaked on day 7 in nontreated allografts, were significantly decreased in AG-treated allografts. AG treatment decreased the number of apoptotic cells and lowered the ratio of the apoptotic cardiac myocytes in contrast to that of the apoptotic infiltrating cells. DNA laddering was clearly detected in nontreated allografts but was suppressed in AG-treated allografts. Inhibition of NO production by AG prolonged murine cardiac allograft survival. The decrease in intra-graft apoptotic activity paralleled histological improvement. Cardiac myocyte death which occurs through an apoptotic process mediated by NO contributes to graft failure during acute cardiac rejection. PMID- 11041101 TI - Anti-CD2 monoclonal antibodies prevent the induction of experimental autoimmune myocarditis. AB - We investigated the effect of a monoclonal antibody against CD2 molecules (OX34) in preventing the induction of experimental autoimmune myocarditis (EAM) induced by immunizing Lewis rats with cardiac myosin. Administration of OX34 before immunization, on Days -6, -4, -2 and 0, completely prevented EAM. On the other hand, treatment with OX34 just before the appearance of myocardial lesions, on Days 9, 11, 13 and 15, had only a partial effect in preventing the disease. Flow cytometric analysis of lymph node cells showed that CD3+ T cells were immediately depleted with the administration of OX34 but had largely recovered on Day 21. Lymph node cells in OX34-treated rats had no proliferative responses to cardiac myosin-rod, but the proliferation was restored when recombinant IL-2 was added. Ultimate production of the anti-myosin antibody was not inhibited by the treatment with OX34. These results suggest that the prevention of EAM by administering the anti-CD2 monoclonal antibody OX34 resulted from T cell depletion during the induction phase, and might in addition result from T cell anergy of Th1, but not Th2 cells. PMID- 11041102 TI - A case of acute myocardial infarction associated with topical use of minoxidil (RiUP) for treatment of baldness. AB - A 45-year-old Japanese man with paroxysmal atrial fibrillation (AF) developed acute anteroseptal myocardial infarction (MI). He had used 1% topical minoxidil (RiUP) once a day for 4 months before the onset of MI for treatment of baldness. Coronary angiography demonstrated severe stenosis at the proximal portion of the left anterior descending coronary artery with a tilling defect. Electrocardiographic monitoring revealed paroxysmal AF and sinus bradycardia with sinus arrests, suggestive of sick sinus syndrome. Topical minoxidil is now widely used for the treatment of male pattern baldness. Although it may be difficult to relate topical use of minoxidil to myocardial ischemia, a greater awareness of its toxicity will be necessary, and patients with cardiovascular disorders should be excluded from the therapy. PMID- 11041103 TI - Papillary fibroelastoma of mitral valve: a rare cause of transient ischemic attack in the young. AB - We report the case of a young Turkish man with a transient ischemic attack secondary to a rare cardiac tumor, papillary fibroelastoma. The tumor was diagnosed by 2-dimensional echocardiography and treated surgically. PMID- 11041104 TI - Marked and prompt hemodynamic improvement by carperitide in refractory congestive heart failure due to dilated cardiomyopathy. AB - A 16-year-old Japanese male diagnosed with congestive heart failure (CHF) due to dilated cardiomyopathy was treated by conventional intensive treatment such as intravenous infusion of diuretics, catecholamines, and phosphodiesterase inhibitors with vasodilators. However, he developed a low output syndrome with appearances of hyponatremia and hypochloremia. As a consequence, intravenous infusion of carperitide (synthetic atrial natriuretic peptide) was added to the therapy. Thereafter his symptoms and hemodynamic parameters promptly and dramatically improved without significant diuresis, and this amelioration persisted for more than 20 days without drug intolerance. This outcome suggests that use of carperitide may be a safe and efficacious means to reduce cardiac preload without hypotension and tachycardia in patients with refractory CHF in whom intensive treatment has already been performed. PMID- 11041105 TI - Isolated right atrial tear following blunt chest trauma: report of three cases. AB - Blunt chest trauma causing isolated right atrial tear and cardiac tamponade in three patients is reported. All three patients presented with hypotension, elevated central venous pressure and altered consciousness. Echocardiographic examination demonstrated pericardial effusion in all three cases. All three patients underwent operation with a median sternotomy approach without using cardiopulmonary bypass. At operation, two patients had one tear in the right atrium, the other had two tears in the right atrium. All three patients recovered uneventfully. Early use of echocardiography to detect the presence of hemopericardium and cardiac tamponade in patients with suspected atrial rupture following blunt chest trauma is advocated. PMID- 11041106 TI - Use of bisphosphonates in skeletal metastasis. AB - At present, there is sufficient evidence to propose practice guidelines that would include the use of bisphosphonates in the management of hypercalcemia, in breast cancer with bone metastases and multiple myeloma. Future research should concentrate on investigating the adjuvant use of bisphosphonates in breast cancer, particularly in order to find out the adequate target groups. Phase III studies comparing the old and new generation bisphosphonates are important as well as trials comparing the other palliative regimens with bisphosphonates. A widespread use of bisphosphonates would have a major impact on drug budgets. Does the cost of achieved palliation represent the optimal use of resources when compared with other possible options for palliation? This issue has not become easier with the emerging new expensive regimens in oncology. An economical analysis, ideally in the setting of randomized trials, is needed. PMID- 11041107 TI - Axillary lymph node dissection in breast cancer--current status and controversies, alternative strategies and future perspectives. AB - Axillary lymph node dissection (ALND) has traditionally been considered as a standard procedure in the surgical management of patients with breast cancer. The goals of ALND in breast cancer surgery are: (a) to provide accurate prognostic information, (b) to maintain local control of the disease in the axilla and (c) to provide a rational basis for decisions about adjuvant therapy. Although controversial, ALND may also be associated with a small therapeutic benefit. Recently, the question of whether ALND is needed for every patient with invasive breast cancer has been the subject of ongoing debate in the literature. This is mainly due to the widespread use of adjuvant systemic therapy for patients with node-negative breast cancer and to the increasingly frequent detection of small invasive cancers by mammographic screening; the majority of these patients have negative axillae. Sentinel lymph node (SLN) biopsy is a new, promising, minimally invasive procedure, which accurately predicts nodal status with minimal morbidity, and reserves ALND for patients with positive SLN biopsies. However, this method is still investigational. Partial (levels I and II) ALND remains the gold standard in the surgical management of patients with breast cancer. PMID- 11041108 TI - Information needs and preferences for participation in treatment decisions among Swedish breast cancer patients. AB - Patient participation in treatment decisions presupposes well-informed patients. The purpose of this study was to determine Swedish breast cancer patients' information needs and their preferences for participation in treatment decisions. Patients (n = 201) were interviewed on nine categories of information and five patient roles, using paired comparisons. Patients gave priority to facts about disease and treatment (chances of cure, stage of disease, treatment options). A collaborative role in treatment decisions was preferred by 87% of the patients. Most patients (56%) preferred a passive form of collaboration: I prefer that my doctor makes the final decision about my treatment but seriously considers my opinion. Younger and better educated patients tended to prefer a more active role. Many patients wanted to be more active (20%) and some more passive (8%) than they actually were. Patients gave priority to disease-specific information, but this reflected needs other than taking control of treatment decisions. PMID- 11041109 TI - Health-related quality of life measured by the EORTC QLQ-C30--reference values from a large sample of Swedish population. AB - The EORTC QLQ-C30 Health-Related Quality of Life (HRQOL) questionnaire was completed by 3069/3919 (78%) of a random sample of the Swedish population aged 18 79 years. The aims of the study were to provide normative data on the questionnaire and to investigate differences in HRQOL with respect to age, gender, sociodemographic characteristics and reported chronic health problems. Women had lower scores than men on all but one of the EORTC QLQ-C30 subscales and reported more chronic health problems. The oldest respondents (70-79 years) had a greater degree of impaired HRQOL than the other age groups, with one exception, 'Emotional functioning', in which they scored higher. Unemployed respondents reported poorer HRQOL than employed respondents. Higher income was associated with a more positive assessment of HRQOL. The results of the study present reference values for EORTC QLQ-C30 Version 3 questionnaire and clarify the influence of factors which should be taken into account when planning studies of HRQOL. PMID- 11041110 TI - Residual mass in aggressive lymphoma--does size, measured by computed tomography, influence clinical outcome? AB - Residual masses are frequently found in patients with aggressive lymphomas, following therapy. A study was undertaken to determine whether initial tumour size, changes during treatment, or size of the residual mass could provide prognostic information. Computed tomography (CT) examinations were carried out before, midway and after completion of chemotherapy in 37 patients with aggressive lymphoma with residual mass after treatment. The tumours were measured for both the greatest diameter sizes and area. The size of the residual mass correlated with the tumour size at diagnosis. Neither a large tumour size before treatment nor a large residual mass after treatment correlated with an increase in rate of relapse. The initial tumour reduction (measured after completion of half of the planned chemotherapy) was less pronounced in relapsing patients compared to relapse-free patients. Using a cut-off level of 70% tumour reduction (measured after completion of half of the planned chemotherapy), 66% of patients with a tumour reduction of < 70% relapsed, compared with 22% (p < 0.05) in those with more marked tumour regression. PMID- 11041111 TI - Correlative study of preoperative transthoracic core cutting needle biopsy of focal thoracic lesions and thoracotomy findings. AB - We conducted a retrospective study to determine the clinical utility of percutaneous core needle biopsy (PCNBx) in 36 patients with peripheral focal chest lesions who later underwent thoracic surgery for diagnostic or therapeutic purposes. PCNBx provided adequate material in 31/36 cases, giving an overall sample yield of 86.1%. PCNBx diagnosis was confirmed by surgery in 27/31 patients, giving a sensitivity of 91.6% and a specificity of 87.5%. In 4 patients, the lesions were misdiagnosed by PCNBx. In 5 patients with benign processes, surgical intervention could have been avoided, according to PCNBx results. The rate of PCNBx-induced pneumothorax was 11%. Radiologically guided PCNBx is an easy and safe procedure that can provide important preoperative diagnostic information and can circumvent the need for exploratory diagnostic surgery in cases of benign lesions. PCNBx also allows better preoperative planning in cases of malignancy. PMID- 11041112 TI - Heterodimerization of Bcl-2 and Bcl-X(L) with Bax and Bad in colorectal cancer. AB - The rate of cell loss owing to apoptosis is mediated by competitive dimerization with selective pairs of cell death antagonists (Bcl-2, Bcl-X(L)) and agonists (Bax, Bad). The aim of this study was to investigate which Bcl-2 family dimers had a critical factor in colorectal cancer. We analyzed the expression of Bcl-2, Bcl-X(L), Bax, and Bad in normal-appearing mucosa and colorectal tumor tissues by Western blotting after immunoprecipitation. Compared with the ratio of Bax-Bcl 2/total Bax in normal mucosa, the ratio was significantly reduced in tumors (p = 0.02). In this series, the low ratio of Bad-Bcl-2/total Bcl-2 was associated with advanced tumor stages (p = 0.02). A reduced heterodimerization of Bax with Bcl-2 may contribute to the development of colorectal cancer. The heterodimerization of Bad with Bcl-2 may be repressed in advanced tumor tissues, and may contribute to tumor growth in colorectal cancer. PMID- 11041113 TI - Feasibility and preliminary results of intensive chemotherapy and extensive irradiation in selected patients with limited small-cell lung carcinoma--results of three consecutive phase II programs. AB - We report the results of three consecutive programs combining initial intensive chemotherapy and radiotherapy in the treatment of patients with limited small cell lung cancer (SCLC). The objective was to test the feasibility and the effect of high-dose chemotherapy and three thoracic irradiation programs on survival and patterns of relapse. Forty-two patients with limited SCLC were enrolled. All patients received high-dose chemotherapy (vindesine, etoposide, doxorubicin, cisplatin and cyclophosphamide or ifosfamide). In the SC 84 program, chest and brain radiotherapy was delivered during each course of chemotherapy, with a complementary irradiation after chemotherapy. In the SC 86 and SC 92 programs, patients received chemotherapy followed by thoracic irradiation and prophylactic brain and spinal axis radiotherapy. At the end of treatment, 40 patients (95%) were in complete response. During chemotherapy, high levels of toxicity were noted. All patients had grade IV hematological toxicities. The extra hematological toxicities were digestive (grade III: 21%; grade IV: 7%) and hepatic (grades III and IV: 14%). During irradiation, patients presented digestive, pulmonary and hematological toxicities. Five patients developed late toxicities and a second malignancy was observed in 4 patients. The 2- and 5-year survival rates for all patients were 51% and 27%, respectively. Despite the marked toxicity of the initial intensive chemotherapy, the treatments are tolerable and effective in the control of extra-thoracic micrometastases, whereas they are less effective for thoracic primary tumor. PMID- 11041114 TI - Lactate dehydrogenase isoenzyme 1 is the most important LD isoenzyme in patients with testicular germ cell tumor. AB - We examined the clinical utility of serum lactate dehydrogenase (LD) isoenzyme catalytic concentrations in 58 patients with testicular germ cell tumors (TGCT) (13 with seminoma and 45 with non-seminomatous tumors). Twenty-one patients with no evidence of disease (NED) all had serum LD isoenzyme 1 catalytic concentrations (LD-1) and LD-1/LD fractions below the upper limit of the reference values (ULR). LD-1 and the LD-1/LD fraction discriminated significantly between evidence of disease (ED) and NED (p = 0.00009 and p = 0.028, respectively, Mann Whitney U-test). Twenty of the 37 patients with ED had raised values of LD-1. The 17 patients with an LD-1 < 1.0 x ULR had a better survival than the 10 patients with LD-1 between 1.0 and 2.9 x ULR, the 7 with LD-1 between 3.0 and 5.9 x ULR, and the 3 patients with LD-1 > 6.0 x ULR (p = 0.006, log-rank test, chi2 test for trend)). Twenty-three patients with an LD-1/LD fraction < or = 0.25 had a better survival than the 14 with an LD-1/LD fraction > 0.25 (p = 0.013). Nineteen patients with LD-5 < 105 U/L and the 15 with LD-5 > 105 U/L had a similar rate of survival (p = 0.85). Our findings add to the evidence showing LD-1 in preference to LD as a serum tumor marker of TGCT. PMID- 11041115 TI - Urinary excretion of platinum in chemotherapy-treated long-term survivors of testicular cancer. AB - As the majority of patients with metastatic testicular cancer are cured by cisplatin-based chemotherapy and can expect an additional life span of around 50 years, late toxicity is of particular relevance. Urine and serum concentrations of platinum were determined by voltammetry in 37 patients at 5.3 to 16.8 years after cisplatin-based chemotherapy. Urinary excretion and serum levels of platinum were 100 to 1000 times higher in patients than in unexposed controls. There may be an association between platinum storage and endocrinologic and metabolic late sequelae, as well as a risk of second cancer. However, further research is necessary to clarify the biological relevance of long-term storage of platinum. PMID- 11041116 TI - Simultaneous radiochemotherapy versus concomitant boost radiation for advanced inoperable head and neck cancer. AB - In this prospective, non-randomized study we compare the results of simultaneous radiochemotherapy (RCT) with those of concomitant boost treatment (CBT) in advanced head and neck cancer. From January 1993 to March 1999, 77 patients were treated with cisplatin, 5-FU, and 70.2 Gy (accelerated split-course); from January 1995 to March 1999, a further 33 patients received CBT to a total dose of 72 Gy. Toxicities were prospectively recorded according to RTOG/EORTC criteria. Acute and subacute reactions did not differ significantly. Severe late effects (III/IV) remained anecdotal (one fistula). Therapy-associated mortalities were 3%(RCT) vs. 0% (CBT), most tumors responding well to therapy (CR + PR: RCT: 72%, CBT: 63%). The 2-year probabilities for freedom from locoregional progression amounted to 42% (RCT) and 31% (CBT); p > 0.05. Tumor-specific 2-year survival amounted to 40% (RCT) and 34% (CBT); p > 0.05. Both of the treatment concepts yield high remission rates with moderate toxicities. Nevertheless, median time to recurrence is still fairly short. We could not find any differences for local control and survival. For patients who are not able to complete the full three courses of radiochemotherapy, the concomitant boost schedule presents a good alternative. PMID- 11041117 TI - Hypopharyngeal squamous cell carcinoma--treatment results in 138 consecutively admitted patients. AB - The aim of this study was to evaluate the results of the initial and salvage treatment for hypopharyngeal carcinoma. The study was conducted in 1963 to 1991 and included 138 patients (38 females (28%) and 100 males (73%)). Most of the tumours originated in the piriform sinus (86%). Tumour stage distribution was T1: 20%, T2: 27%, T3: 37% and T4: 17% and nodal stage distribution was N0: 45%, N1: 25%, N2: 10%, and N3: 20%. Primary treatment was delivered with curative intent in 124 out of 138 cases (90%). Treatment failure was noted in 98 patients, with 55% recurrence in T-position, 39% in N-position, and 14% at distant metastases sites. Salvage surgery was successful in 9 out of 32 patients. The overall 5-year locoregional tumour control, cause-specific and overall survival rates were 20%, 25% and 19%, respectively. Univariate actuarial analysis showed that T- and N stage, clinical stage, tumour size and well-differentiated tumours were significant prognostic parameters. A Cox multivariate analysis showed that only the T- and N-stages were independent prognostic factors. In conclusion, the prognosis for advanced hypopharyngeal carcinoma is extremely poor and the meagre results with conventional radiotherapy alone indicate that other treatment modalities should be introduced in the management of this disease. PMID- 11041118 TI - Raltitrexed-related pulmonary toxicity. PMID- 11041119 TI - Nutrition and mortality: the Finnish experience. AB - This paper reviews the Finnish experience concerning nutrition and mortality. In the early 70's major activities were started to change national nutritional habits, particularly in view of reducing the high mortality of cardiovascular diseases. At the same time a comprehensive monitoring system was developed. During the period from 1972 to 1997 major changes took place in national nutrition, in accordance with the set objectives. Associated with this, major reduction was observed in national CVD mortality. The Finnish experience gives great support to the hypothesis that population rates of cardiovascular diseases can be substantially influenced by changes in people's dietary habits. PMID- 11041120 TI - Endothelial dysfunction in hypertension. AB - Endothelial cells release both relaxing and contracting factors that modulate vascular smooth muscle tone and also participate in the pathophysiology of essential hypertension. Endothelium-dependent vasodilation is regulated primarily by nitric oxide but also by an unidentified endothelium-derived hyperpolarizing factor and by prostacyclin. Endothelium-derived contracting factors include endothelin-1, vasoconscrictor prostanoids, angiotensin II and superoxide anions. Under physiological conditions, there is a balanced release of relaxing and contracting factors. The balance can be altered in cardiovascular diseases such as hypertension, atherosclerosis, diabetes and other conditions, thereby contributing to further progression of vascular and end-organ damage. In particular, endothelial dysfunction leading to decreased bioavailability of nitric oxide impairs endothelium-dependent vasodilation in patients with essential hypertension and may also be a determinant for the premature development of atherosclerosis. Different mechanisms of reduced nitric oxide activity have been shown both in hypertensive states and several cardiovascular diseases, and endothelial dysfunction is likely to occur prior to vascular dysfunction. Thus, the strategies currently used to improve endothelial dysfunction may result in decreased morbidity and mortality in hypertensive patients. PMID- 11041121 TI - Porcine coronary imaging in vivo by optical coherence tomography. AB - OBJECTIVE: A high-resolution coronary artery imaging modality has the potential to address important diagnostic and management problems in cardiology. Optical coherence tomography (OCT) is a promising new optical imaging technique with a resolution of approximately 10 microm. The purpose of this study was to use a new OCT catheter to demonstrate the feasibility of performing OCT imaging of normal coronary arteries, intimal dissections, and deployed stents in vivo. METHODS AND RESULTS: Normal coronary arteries, intimal dissections, and stents were imaged in five swine with OCT and compared with intravascular ultrasound (IVUS). In the normal coronary arteries, visualization of all of the layers of the vessel wall was achieved with a saline flush, including the intima which was not identified by IVUS. Following dissection, detailed layered structures including intimal flaps, intimal defects, and disruption of the medial wall were visualized by OCT. IVUS failed to show clear evidence of intimal and medial disruption. Finally, the microanatomic relationships between stents and the vessel walls were clearly identified only by OCT. CONCLUSIONS: In this preliminary experiment, we have demonstrated that in vivo OCT imaging of normal coronary arteries, intimal dissections, and deployed stents is feasible, and allows identification of clinically relevant coronary artery morphology with high-resolution and contrast. PMID- 11041122 TI - Are the WOSCOPS clinical and economic findings generalizable to other populations? A case study for Belgium. The WOSCOPS Economic Analysis Group. West of Scotland Coronary Prevention Study. AB - AIMS: As the West of Scotland Coronary Prevention Study (WOSCOPS) was conducted in Scotland, a country well-known for its high cardiovascular risk, the generalizability of its findings on pravastatin's clinical and economic effects has been questioned. This study examines the legitimacy of this concern, using Belgium as a case study. METHODS AND RESULTS: Local information on the prevalence and clustering of risk factors in individual patients was used in a risk equation to estimate the reference risk in Belgium. In contrast to prevailing beliefs, this risk was shown to coincide with the trial population's risk. As the relative risk reduction documented in a trial should apply across populations, the health benefits observed in WOSCOPS can clearly be extrapolated. This information in combination with local costs was then used to assess the economic efficiency of primary prevention with pravastatin in Belgium by means of a previously developed model. In parallel with the original estimates for the United Kingdom, the cost effectiveness ratios remain well within the range of what is considered strong to moderate evidence for adoption and appropriate utilization, over a wide range of input values. CONCLUSION: This study demonstrates that the clinical and economic findings from WOSCOPS can indeed be generalized to other populations. PMID- 11041124 TI - Six months clinical, angiographic, and IVUS follow-up after PTFE graft stent implantation in native coronary arteries. AB - INTRODUCTION: Restenosis remains a problem even after stent implantation. An important breakthrough could be the use of graft stents, functioning as a mechanical barrier between the blood flow and the vessel wall, and possibly inducing less restenosis by more limited hyperplasia and minimal transgraft tissue penetration. OBJECTIVE: To assess the acute and 6 months clinical, angiographic and IVUS results of a new balloon expandable coronary polytetrafluoroethylene (PTFE) graft stent (Jomed). METHOD: Ten patients with a short (< or = 15 mm length) de novo proximal stenosis in a large (> or = 3 mm diameter) coronary artery were treated by elective implantation of a graft stent (19 mm stent, 15 mm graft). Clinical assessment, quantitative coronary angiography (QCA) and intracoronary ultrasound (IVUS) were performed before, immediately after and 6 months after implantation. A stress test was also done at 6 months. RESULTS: The coronary arteries treated were: RCA in 7 patients, LCX in 2 patients, LAD in 1 patient. Mean balloon size was 3.7 mm diameter, and mean inflation pressure was 18 atm (min. 12, max. 23). Additional stenting was needed in 3 patients. Two patients showed a minimal rise in CK (< 250 IU/l) and 1 patient needed a transfusion. No patient experienced a (sub)acute nor late thrombosis. As shown in the table, no restenosis was seen in the body of the graft stent. In 2 patients a restenosis was detected in the proximal and/or distal parts of the stent which are not covered by the graft. In 1 patient a restenosis was found outside the stent. All patients remained asymptomatic with a negative stress test at 6 months follow-up (FU). [table in text] CONCLUSIONS: A graft stent could indeed reduce the restenosis rate after stenting, in the part of the stent covered by the graft, but the uncovered distal and proximal parts are the weak points in this type of stent. For this reason, technical ameliorations in the construction of this graft stent are needed, e.g. a complete coverage of the stent by the PTFE graft and less rigidity of the stent causing reduced vessel trauma at the edges of the stent during implantation. PMID- 11041123 TI - Alterations in the fibrinolytic system components during acute myocardial infarction. AB - The purpose of this study was to evaluate the changes in tissue-plasminogen activator (t-PA), plasminogen activator inhibitor - type 1 (PAI-1) and D-dimer (DD) antigen plasma levels in acute myocardial infarction (AMI) patients after thrombolytic therapy with two different thrombolytic agents, rt-PA or acetyl streptokinase and to find out any correlation between the plasma t-PA, PAI-1 and DD levels with the infarct size as it is estimated from the peak of serum CPK levels. The plasma antigen levels of t-PA, PAI-1 and DD were measured by the enzyme immunoassay method (Stago), in 57 consecutive patients (M = 46, F = 11, mean age 55.6 +/- 8.8 years) and in 25 normal subjects (M = 18, F = 7, mean age 54.0 +/- 5.5 years). In 47 out of the 57 patients who were treated successfully with 100 mg of rt-PA (26 patients) or with 1.5 MU 21 of acetyl-streptokinase, as well as in 10 patients who were not treated, samples were obtained again 4 and 24 hours after the end of thrombolytic therapy or admission, respectively. During the acute phase of myocardial infarction the t-PA, PAI-1 and DD antigen plasma levels were significantly higher than in healthy people. There were no significant changes in the t-PA, PAI-1 and DD plasma levels of the patients who were not treated with a thrombolytic agent. We found a significant elevation of t PA (p < 0.001), PAI-1 (p < 0.05) and DD (p < 0.001) after 4 hours in comparison with the baseline (at presentation, before therapy). After 24 hours the t-PA and DD plasma levels remained significantly higher (p < 0.001) while the PAI-1 plasma levels returned to the pre-therapy levels. There were no significantly different changes in the t-PA, PAI-1 and DD plasma levels of either group of patients, treated with rt-PA or acetyl-streptokinase while the t-PA and PAI-1 levels were positively correlated with infarct size as estimated from peak serum CPK levels. PMID- 11041125 TI - Surgical treatment of cardiogenic shock due to huge right atrial thromboembolus. AB - An unusual case is reported of thromboembolus in the right atrium associated with cardiogenic shock and multiple pulmonary micro-embolisms. Two-dimensional echocardiograpy demonstrated a large irregular mass in the right atrium floating freely, prolapsing through the tricuspid valve into the right ventricle during diastole, and leading to inflow and outflow obstruction. An emergency operation was performed and the thromboembolic material was successfully extracted from the right atrium without using cardiopulmonary bypass. This exemplary case highlights the benefit of surgical intervention rather than more conservative approaches such as anticoagulation and/or thrombolysis. PMID- 11041127 TI - Left ventricular false aneurysm. AB - False aneurysms of the left ventricle develop after rupture of the ventricular wall in an area of pericardial adhesions. This complication of myocardial infarction is uncommon. Images of a post-infarction false aneurysm are presented. PMID- 11041126 TI - Failure of medicine: evolution of an atrial septal defect to an Eisenmenger syndrome. AB - The atrial septal defect (ASD) is the most commonly diagnosed congenital defect in adults and has a prevalence of 7.5% of all congenital cardiac anomalies. Less invasive imaging techniques, especially transthoracic and transoesophageal echocardiography, provide more accurate diagnostics, resulting in earlier diagnosis and treatment. Despite these opportunities in high-tech countries, medicine may still fail in detecting initially correctable cardiac anomalies. We present a case of 41-year-old woman with an abnormal murmur at childhood that disappeared with time due to the development of an Eisenmenger syndrome. The importance of a complete haemodynamic evaluation in this patient is illustrated. PMID- 11041128 TI - A summary of reported foodborne disease incidents in Sweden, 1992 to 1997. AB - Reports of foodborne disease incidents in Sweden from 1992 to 1997 are summarized. The results are based on reports from the municipal environmental and public health authorities to the National Food Administration and from medical authorities to the Swedish Institute for Infectious Diseases Control. A total of 555 incidents, of which 84% were outbreaks, were reported, involving 11,076 ill people. In 66% of the incidents, no disease agent was determined. Bacterial agents were implicated in 25% and viruses in 8% of the incidents. Calicivirus was the most reported agent both in terms of incidents and cases. Mixed dishes was the food category most often implicated in outbreaks, and smorgasbord and casserole or stews were the subcategories that caused the most cases. The place of consumption was unknown in 8% of the incidents. In about 60% of the incidents, the implicated food was consumed in commercial food establishments; in approximately 20% of incidents, it was consumed at home. The average annual incidence of reported foodborne disease in Sweden was estimated to be 21 cases per 100,000. The average annual incidence of reported foodborne salmonellosis and campylobacteriosis was estimated to be 2.0 and 0.6 cases per 100,000, respectively. The awareness and motivation to report foodborne diseases need to be improved, but additional sources of information are needed to counteract some of the limitations of reporting discussed in this work. PMID- 11041129 TI - Evaluation of safe food-handling instructions on raw meat and poultry products. AB - Every year in the United States, millions of people become ill, thousands of people die, and substantial economic costs are incurred from foodborne diseases. As a measure to prevent foodborne diseases, since July 1994, the U.S. Department of Agriculture has required that safe food-handling labels be placed on retail packages of raw or partially cooked meat and poultry products. Through selected states' Behavioral Risk Factor Surveillance System (BRFSS) interviews, survey data were collected to determine the proportion of adults aware of the label and adults who reported changing their raw meat-handling practices because of the label. Fifty-one percent of the 14,262 respondents reported that they had seen the label. Of these, 79% remembered reading the label, and 37% of persons who reported that they had seen and read the label reported changing their raw meat preparation methods because of the label. Women were more likely than men to have read the label, as were persons who are at least 30 years of age compared to younger adults (P < 0.05). Both label awareness and risky food-handling behaviors increased with education and income, suggesting that safe food-handling labels have limited influence on consumer practices. Our results also suggest that the labels might be more effective in discouraging cross-contamination than in promoting thorough cooking practices. We suggest that the label is only one component among many food safety education programs that are needed to inform consumers about proper food-handling and preparation practices and to motivate persons who have risky food-handling and preparation behaviors to change these behaviors. PMID- 11041130 TI - Interventions for the reduction of Salmonella Typhimurium DT 104 and non-O157:H7 enterohemorrhagic Escherichia coli on beef surfaces. AB - A study was conducted to determine if slaughter interventions currently used by the meat industry are effective against Salmonella Typhimurium definitive type 104 (DT 104) and two non-O157:H7 enterohemorrhagic Escherichia coli (EHEC). Three separate experiments were conducted by inoculating prerigor beef surfaces with a bovine fecal slurry containing Salmonella Typhimurium and Salmonella Typhimurium DT 104 (experiment 1), E. coli O157:H7 and E. coli O111:H8 (experiment 2), or E. coli O157:H7 and E. coli O26:H11 (experiment 3) and spray washing with water, hot water (72 degrees C), 2% acetic acid, 2% lactic acid, or 10% trisodium phosphate (15 s, 125 +/- 5 psi, 35 +/- 2 degrees C). Remaining bacterial populations were determined immediately after treatments (day 0), after 2 days of aerobic storage at 4 degrees C, and after 7, 21, and 35 days of vacuum-packaged storage at 4 degrees C. In addition to enumeration, confirmation of pathogen serotypes was performed for all treatments on all days. Of the interventions investigated, spray treatments with trisodium phosphate were the most effective, resulting in pathogen reductions of >3 log10 CFU/cm2, followed by 2% lactic acid and 2% acetic acid (>2 log10 CFU/cm2). Results also indicated that interventions used to reduce Salmonella Typhimurium on beef surfaces were equally effective against Salmonella Typhimurium DT 104 immediately after treatment and again after long-term, refrigerated, vacuum-packaged storage. Similarly, E. coli O111:H8 and E. coli O26:H11 associated with beef surfaces were reduced by the interventions to approximately the same extent as E. coli O157:H7 immediately after treatment and again after long-term, refrigerated, vacuum-packaged storage. It was also demonstrated that phenotypic characterization may not be sufficient to identify EHECs and that the organisms should be further confirmed with antibody- or genetic-based techniques. Based on these findings, interventions used by the meat industry to reduce Salmonella spp. and E. coli O157:H7 appear to be effective against DT 104 and other EHEC. PMID- 11041131 TI - Protective effect of Enterococcus faecium J96, a potential probiotic strain, on chicks infected with Salmonella Pullorum. AB - Enterococcus faecium J96 was isolated from a healthy free-range chicken and it inhibited Salmonella Pullorum, in vitro, due to its lactic acid and bacteriocin production. In vivo assays were carried out with 30-h-old broiler chicks. The lactic acid bacteria (approximately 1 x 10(9) cells per chick) were orally administered as preventive and as therapeutic treatments. In the first case they were given to the chicks twice a day for 3 consecutive days. In the second case the lactic bacteria were administered in the same way after a 24-h challenge by Salmonella Pullorum (in both instances the salmonella dose was 1 x 10(5) cells per chick). Cecal contents, liver, and spleens were analyzed and liver and spleen fragments were also fixed in formaldehyde (pH 7.00) in order to determine salmonella translocation. The chickens that were preventively treated with E. faecium J96 survived the Salmonella Pullorum challenge. Those that were infected on the first day and then inoculated with lactic bacteria died 4 days later. Salmonellae were isolated from their livers and spleens. From these results we may conclude that E. faecium J96 can protect newly hatched chicks from Salmonella Pullorum infection but cannot act as a good therapeutic agent. PMID- 11041132 TI - Surface application of lysozyme, nisin, and EDTA to inhibit spoilage and pathogenic bacteria on ham and bologna. AB - A study was conducted to determine if the effectiveness of an antimicrobial treatment for cooked ham and bologna would be increased or maintained when applied in a surface coating. Cooked 10-g disks of ham and bologna sausage received one of three treatments: no coating (control), coating with 0.2 g of 7% (wt/vol) gelatin gel (gel-control), or coating with 0.2 g of 7% gelatin gel containing 25.5 g/liter of lysozyme-nisin (1:3) plus 25.5 g/liter of EDTA (gel treated). The samples were then inoculated with one of six test organisms: Brochothrix thermosphacta, Escherichia coli O157:H7, Lactobacillus sakei, Leuconostoc mesenteroides, Listeria monocytogenes, or Salmonella Typhimurium. Inoculated samples were vacuum packed and stored at 8 degrees C for 4 weeks. The antimicrobial gel treatment had an immediate bactericidal effect up to 4 log CFU/cm2 on the four gram-positive organisms tested (B. thermosphacta, Lactobacillus sakei, Leuconostoc mesenteroides, and Listeria monocytogenes) and inhibited the growth of these organisms during the 4 weeks of storage. The antimicrobial gel treatment also had a bactericidal effect on the growth of Salmonella Typhimurium during storage. The numbers of E. coli O157:H7 on ham were reduced by 2 log CFU/cm2 following treatment with both antimicrobial-containing and non-antimicrobial-containing gels during the 4-week storage period. No effect was observed on the growth of E. coli O157:H7 on bologna. PMID- 11041134 TI - Influence of traditional brine washing of smear Taleggio cheese on the surface spreading of Listeria innocua. AB - The influence of a traditional procedure of washing of smear Taleggio cheese on surface spreading of Listeria innocua was studied. This practice is carried out during ripening to remove molds, to select the surface microflora, and to control the ripening process. One cheese, both of 2 (i) and 4 (ii) weeks of ripening, was surface-inoculated with approximately 3 log CFU of L. innocua per entire cheese surface. The inoculated cheeses and others of the same age were weekly washed with brine solution. Listeria was spread both on the surface of the inoculated cheese and on the other cheeses, and it was also found in the brines and on the wooden boxes where the cheeses were ripened. The time of ripening when contamination occurs influenced the behavior of Listeria. At the moment of contamination, the smear surface microflora of (i) cheese was approximately 2 log CFU/g higher than of (ii) cheese. Listeria inoculated on 2-week-ripened cheese was able to colonize the entire surface of the cheese and to cross-contaminate the other cheeses. On the contrary, Listeria inoculated on a 4-week-ripened cheese was partially spread on the surface of the originally inoculated cheese, and the transfer of contamination by the washing procedure was restrained. Because a random distribution of Listeria on cheese surface was observed, the importance of the mode of sampling was discussed. Because of the lack of critical control points during ripening of Taleggio cheese, the Listeria hazard needs to be controlled by taking appropriate control measures to break off the contamination cycle (cheese --> brine --> wooden boxes --> cheese). PMID- 11041133 TI - Differentiation between types and strains of Clostridium botulinum by riboprinting. AB - The ability of automated ribotyping to differentiate between major types and individual strains of Clostridium botulinum was tested using the Qualicon Riboprinter Microbial Characterization System. Pure spores of C. botulinum type A, proteolytic type B, nonproteolytic type B, and type E strains were inoculated onto modified anaerobic egg yolk agar and incubated 24 h at 35 degrees C. Plates were rinsed with buffer (2 mM Tris + 20 mM EDTA) to remove vegetative cells that were heated for 10 min at 80 degrees C, treated with a lysing agent, and ribotyped in the Qualicon Riboprinter utilizing the enzyme EcoRI. Riboprint patterns were obtained for 30 strains of the four major types of C. botulinum most commonly involved in human foodborne botulism. Proteolytic strains yielded the best and most consistent results. Fifteen ribogroups were identified among the 31 strains tested. Interestingly, in two cases, a single ribogroup contained patterns from isolates belonging to evolutionarily distinct Clostridium lineages. This degree of differentiation between strains of C. botulinum may be useful in hazard analysis and identification, hazard analysis and critical control point monitoring and validation, environmental monitoring, and in inoculation studies. PMID- 11041135 TI - Antimicrobial effect of rosemary extracts. AB - A rosemary extract commercially exploited (Oxy'less) as an antioxidant of lipids in foods was dissolved in ethanol (100 mg/ml), and the solution was tested against foodborne microorganisms. For gram-positive bacteria, the MIC of the ethanolic solution was 1% for Leuconostoc mesenteroides, 0.5% for Listeria monocytogenes, 0.5% for Staphylococcus aureus, 0.13% for Streptococcus mutans, and 0.06% for Bacillus cereus. It slowed the growth of Penicillium roquefortii and Botrytis cinerea. Up to 1% of the ethanolic solution had no activity on the gram-negative bacteria Escherichia coli, Salmonella Enteritidis, and Erwinia carotovora and on the yeasts Rhodotorula glutinis and Cryptococcus laurentii. Antibacterial activity of the rosemary extract was strongly influenced by the composition of the media. The MIC was reduced by low pH, high NaCl contents, and low temperatures. Low pH and high NaCl concentration had a synergistic effect on the MIC of the rosemary extract for S. aureus. Lipids, surface-active agents, and some proteins decreased its antibacterial activity, whereas pectin had no effect. The inhibitory effect was little modified by heat treatment (100 degrees C). The natural microflora of pasteurized zucchini broth was inhibited by 0.5% of the rosemary extract. The antibacterial activity was linked to the compounds extracted with hexane, which are presumably phenolic diterpenoids. PMID- 11041137 TI - Heat resistance of Alicyclobacillus acidocaldarius in water, various buffers, and orange juice. AB - The effect of the pH or the composition of the heating medium and of the sporulation temperature on the heat resistance of spores of a thermoacidophilic spore-forming microorganism isolated from a dairy beverage containing orange fruit concentrate was investigated. The species was identified as Alicyclobacillus acidocaldarius. The spores showed the same heat resistance in citrate-phosphate buffers of pH 4 and 7, in distilled water, and in orange juice at any of the temperatures tested (D120 degrees C = 0.1 min and z = 7 degrees C). A raise in 20 degrees C in the sporulation temperature (from 45 to 65 degrees C) increased the heat resistance eightfold (from D110 degrees C = 0.48 min when sporulated at 45 degrees C to 3.9 min when sporulated at 65 degrees C). The z values remained constant for all sporulation temperatures. The spores of this strain of A. acidocaldarius were very heat resistant and could easily survive any heat treatment currently applied to pasteurize fruit juices. PMID- 11041136 TI - Effect of conjugated bile salts on antibiotic susceptibility of bile salt tolerant Lactobacillus and Bifidobacterium isolates. AB - Virtually every antibiotic may cause in vivo alterations in the number, level, and composition of the indigenous microbiotae. The degree to which the microbiotae are disturbed depends on many factors. Although bile may augment antibiotic activity, studies on the effect of bile on the antibiotic susceptibility of indigenous and exogenous probiotic microorganisms are lacking. It was against this background that the antibiotic susceptibility of 37 bile salt tolerant Lactobacillus and 11 Bifidobacterium isolates from human and other sources was determined in the presence of 0.5% wt/wt oxgall (conjugated bile salts). Oxgall did not affect the intrinsic resistance of lactobacilli to metronidazole (5 microg), vancomycin (30 microg), and cotrimoxazole (25 microg), whereas it resulted in a complete loss of resistance to polymyxin B (300 microg) and the aminoglycosides gentamicin (10 microg), kanamycin (30 microg), and streptomycin (10 microg) for most strains studied (P < 0.001). Oxgall did not affect the intrinsic resistance of bifidobacteria to metronidazole and vancomycin, whereas polymyxin B and co-trimoxazole resistance was diminished (P < 0.05) and aminoglycoside resistance was lost (P < 0.001). Seven lactobacilli, but no bifidobacteria strain, showed unaltered intrinsic antibiotic resistance profiles in the presence of oxgall. Oxgall affected the extrinsic susceptibility of lactobacilli and bifidobacteria to penicillin G (10 microg), ampicillin (10 microg), tetracycline (30 microg), chloramphenicol (30 microg), erythromycin (15 microg), and rifampicin (5 microg) in a source- and strain-dependent manner. Human strain-drug combinations of lactobacilli (P < 0.05) and bifidobacteria (P < 0.01) were more likely to show no change or decreased susceptibility compared with other strain-drug combinations. The antimicrobial activity spectra of polymyxin B and the aminoglycosides should not be considered limited to gram negative bacteria but extended to include gram-positive genera of the indigenous and transiting microbiotae in the presence of conjugated bile salts. Those lactobacilli (7 of 37) that show unaltered intrinsic and diminished extrinsic antibiotic susceptibility in the presence of oxgall may possess greater upper gastrointestinal tract transit tolerance in the presence of antibiotics. PMID- 11041138 TI - Extension of the shelf life of prawns (Penaeus japonicus) by vacuum packaging and high-pressure treatment. AB - The present study has investigated the application of high pressures (200 and 400 MPa) in chilled prawn tails, both conventionally stored (air) and vacuum packaged. Vacuum packaging and high-pressure treatment did extend the shelf life of the prawn samples, although it did affect muscle color very slightly, giving it a whiter appearance. The viable shelf life of 1 week for the air-stored samples was extended to 21 days in the vacuum-packed samples, 28 days in the samples treated at 200 MPa, and 35 days in the samples pressurized at 400 MPa. Vacuum packaging checked the onset of blackening, whereas high-pressure treatment aggravated the problem. From a microbiological point of view, batches conventionally stored reached about 6 log CFU/g or even higher at 14 days. Similar figures were reached in total number of bacteria in vacuum-packed samples and in pressurized at 200-MPa samples at 21 days. When samples were pressurized at 400 MPa, total numbers of bacteria were below 5.5 log CFU/g at 35 days of storage. Consequently, a combination of vacuum packaging and high-pressure treatment would appear to be beneficial in prolonging freshness and preventing spotting. PMID- 11041139 TI - Visual color and doneness indicators and the incidence of premature brown color in beef patties cooked to four end point temperatures. AB - An interlaboratory study was undertaken to assess the frequency that cooked color of ground beef patties appeared brown at internal temperatures of 52.7 degrees C (135 degrees F), 65.6 degrees C (150 degrees F), 71.1 degrees C (160 degrees F), and 79.4 degrees C (175 degrees F). In general, as internal cooked temperature of the patties increased, the following results were observed in the patties: (i) more brown meat color, (ii) less pink or red juice color, and (iii) more cooked texture. However, brown meat color occurred prematurely at the two lower internal temperatures (57.2 degrees C/135 degrees F and 65.6 degrees C/150 degrees F) that are insufficient to eliminate foodborne pathogens without holding times. The common consumer practice of freezing bulk ground beef, followed by overnight thawing in a refrigerator, led to substantial premature brown color in patties cooked from this product. In addition, at 71.1 degrees C (160 degrees F), recognized to be the lowest temperature for cooking ground beef safely in the home, meat color, juice color, and texture appearance were not fully apparent as doneness indicators. In fact, at no temperature studied did 100% of the patties appear done when evaluated by the criteria of no red or pink in the meat, no red or pink in the juices, or by texture appearance. Patties in this study were evaluated under a set protocol for forming the products, cooking, and viewing under the same lighting conditions. Other preparation conditions are possible and may produce different results. Thus, temperature to which patties have been cooked cannot be judged by color and appearance. This study provided the evidence to support the message to consumers regarding cooking of beef patties of "use an accurate food thermometer and cook beef patties to 160 degrees F (71.1 degrees C)" in place of messages based on consumer judgment of cooked color. PMID- 11041140 TI - The mycobiota of speck, a traditional Tyrolean smoked and cured ham. AB - Speck is a ham specialty product traditionally produced in South Tyrol (Italy) and North Tyrol (Austria) by farmers, butcheries, and meat industries. To date, nothing has been learned about fungi associated with this smoked and cured meat. Therefore, it was the main objective of this study to assess the typical mycobiota of Speck in relation to the different production types and the geographic provenance. A total of 121 Speck samples from North Tyrol and South Tyrol was analyzed. From 63 isolated fungal species, only a few can be regarded as typical colonizers: Eurotium rubrum and Penicillium solitum were the dominating species in all types and parts of Speck (crust, meat, and fat). Eight other Penicillium spp. were relatively frequent. The species diversity increased from industrially produced Speck to products from butcheries and farmers, and it was higher in all types of South Tyrolean products. Among the typical mycobiota, Penicillium verrucosum, Penicillium canescens, and Penicillium commune are known as potentially mycotoxigenic. PMID- 11041141 TI - Comparison of capillary and test tube procedures for analysis of thermal inactivation kinetics of mold spores. AB - Characteristics of capillary and test tube procedures for thermal inactivation kinetic analysis of microbial cells were studied for mold spores. During heating, capillaries were submerged in a water bath and test tubes were held with their caps positioned above the level of the heating medium. Thermal inactivation curves of Aspergillus niger spores in capillaries at around 60 degrees C consisted of a shoulder and a fast linear decline, whereas curves in test tubes consisted of a shoulder, a fast linear decline, and a horizontal tail. There were no significant differences in values of the rate and the delay of fast declines in curves between the procedures. Some experiments were done to clarify the cause for tailing with test tubes. There were no tails with test tubes whose inner walls were not contaminated by A. niger spores, suggesting that tails arise from A. niger spores contaminating the inner walls of test tubes. Temperature of the inner wall at the level of a heating medium was lower than that of the medium. Further, there were no tails for test tubes submerged in the heating medium. These results showed that the reason for survival of contaminants on the upper wall of test tubes was that cells were not subjected to sufficient inactivation temperature. Finally, thermal inactivation curves of A. niger spores in capillaries at various constant temperatures were studied. Curves consisted of a shoulder and a fast linear decline at 57 degrees C and above, whereas curves at below 57 degrees C consisted of a shoulder, a fast linear decline, and a sloping tail. PMID- 11041142 TI - Isolation of Cyclospora oocysts from fruits and vegetables using lectin-coated paramagnetic beads. AB - Published techniques for recovering parasites from fruit and vegetables are generally inadequate, with low and variable recovery efficiencies. Herein, we describe an improved method for analyzing fruit and vegetables for Cyclospora oocysts. The technique includes washing procedures, sonication, and separation using lectin-coated paramagnetic beads. Identification is by microscopy (differential interference contrast and fluorescence). Oocyst recovery efficiencies from mushrooms, lettuce, and raspberries were approximately 12%. Recovery efficiencies from bean sprouts were approximately 4%. Although no significant difference in recovery efficiency could be detected between samples processed using the lectin-coated beads and samples processed without this procedure, distinct advantages were apparent when the lectin-coated beads were used. A considerably smaller, cleaner final volume remained for microscopy, which increases the sensitivity of the technique and reduces operator time. PMID- 11041143 TI - Formation of heterocyclic amines in fried fish fiber during processing and storage. AB - The formation of heterocyclic amines (HAs) in fried fish fiber during processing and storage was studied. Fried fish fiber was prepared by boiling of raw fish, followed by eviscerating, pressing, chopping, and then the fish meat was subjected to frying, during which the various additives such as sugar, soybean sauce, and edible oil were added. The various HAs in fried fish fiber were analyzed by high-performance liquid chromatography with photodiode-array detection. Only four HAs, Norharman, Harman, 2-amino-9H-pyrido[2,3-b]indole, and 2-amino-3-methyl-9H-pyrido[2,3-b]indole were detected in fried fish fiber. The amount of HAs increased with increasing frying temperature. Amino acids might play a more important role for HA formation than reducing sugar during processing of fried fish fiber. During storage, the HAs increased with increasing storage temperature when the fried fish fiber was packed in an aluminum foil bag. However, the relationship between storage temperature and HAs formation was not consistent when the fried fish fiber was packed in a plastic bag. PMID- 11041144 TI - Detection, quantitation, and identification of residual aminopenicillins by high performance liquid chromatography after fluorescamine derivation. AB - A high-performance liquid chromatographic (HPLC) method with fluorescence detection after precolumn fluorescamine derivation was developed to detect residues of two aminopenicillins, amoxicillin (AMPC) and ampicillin (ABPC), in bovine serum. Proteins in serum samples spiked with each of these penicillins were precipitated with sodium tungstate and sulfuric acid, centrifuged, and removed by passage through a C18 solid-phase extraction cartridge. After precolumn treatment of the extraction products of AMPC and ABPC with fluorescamine solution, HPLC analysis with fluorescence spectrophotometric detection at an excitation wavelength of 390 nm and an emission wavelength of 485 nm was performed to identify these products. Two mobile phases were used for residual analysis by the isocratic HPLC system. An ODP column (polyvinyl alcohol bonded with an octadecyl functional group) that can be used with strongly alkaline mobile phases (pH 2.0 to 13) was selected, and the column temperature was set at 40 degrees C. A mobile phase comprising 100-mM K2HPO4 solution and acetonitrile (72:28, vol/vol), which yielded AMPC and ABPC retention times of 4.1 and 7.9 min, respectively, was suitable for detection of residual ABPC but not for residual AMPC because interference was caused by peaks of other extracted substances. When a mobile phase comprising a different ratio of 100-mM K2HPO4 solution and acetonitrile (78:22, vol/vol) was used, the retention times of AMPC and ABPC were 7.3 and 26.3 min, respectively, and both penicillins could be analyzed using this system. The calculated standard curves of the reaction products with both mobile phases were linear, and the correlation coefficients were greater than 0.999. The lower limit of detection was 10 ng/ml for both penicillins. Analysis of extracts from bovine serum spiked with AMPC and ABPC at a concentration of 1 microg/ml yielded recovery rates of 102.2 +/- 5.5% and 79.0 +/- 5.2%, respectively. This detection method may be useful for routine laboratory testing of AMPC and ABPC. PMID- 11041145 TI - Milkborne campylobacter infection in Hungary. AB - In April 1998, an annual 2-day animal farm sale was held in Hodmezovasarhely, where 500 to 600 visitors consumed unpasteurized milk. The first signs of disease began 2 days after the end of the sale. Fifty-two people from a wide age range fell ill, primarily with inflammatory enteritis. These cases included 34 with Campylobacter positivity: 30 with Campylobacter jejuni and 4 with Campylobacter coli. Environmental samples (raw milk, udder swabs, and rectal swabs from 12 cows in the suspected herd) were tested 2 weeks after the first signs of the disease, and two rectal swabs were found to be positive for C. jejuni. Initially, the epidemic seemed to be sporadic and, accordingly, only 26 human and 2 animal Campylobacter isolates were reserved for randomly amplified polymorphic DNA analysis. This comparative analysis verified that fecally contaminated milk was the source of the outbreak. The DNA-banding patterns of 20 C. jejuni isolates (19 human and 1 animal) were identical. The antibiotic susceptibilities of the Campylobacter isolates were determined, and only six C. jejuni (human) isolates, one C. coli (human) isolate, and one C. jejuni (animal) isolate were resistant to tetracycline, both by disk diffusion and by E test (antimicrobial gradient strip for the quantitative determination of susceptibility or resistance of microorganisms). No plasmid was detected in these tetracycline-resistant isolates. The endotoxin production of Campylobacter isolates was examined via the cytopathogenic effect on the Vero cell line. This effect exhibited various degrees of positivity in 19 cases. Only two human C. jejuni isolates displayed + + + + positivity. Both isolates were from patients who had required antibiotic therapy and hospital care. PMID- 11041146 TI - Diminution of Campylobacter colonization in neonatal pigs reared off-sow. AB - Pigs may be a natural reservoir of Campylobacter and can be colonized as early as 24 h after birth. The purpose of the present study was to evaluate what effect early removal of piglets from Campylobacter-positive sows has on Campylobacter prevalence in neonates. In two trials, piglets were removed from sows within 24 h of birth and were reared in nurseries isolated from sows for 21 days. From the neonates rectal swabs were cultured for Campylobacter, and Campylobacter status of the isolated piglets was compared to that of littermates reared on sows. The nurseries consisted of wire-floored farrowing crates that were equipped with heaters and self-feeders. In trial I, the Campylobacter prevalence in nursery reared piglets was 13 of 14 on day 2 and 0 of 14 on day 20. Campylobacter prevalence in the sow-reared piglets was 8 of 9 from days 2 to 20. In trial II, 12 of 29 on day 2, and 5 of 26 on day 20, of the nursery-reared piglets were culture positive for Campylobacter. For the sow-reared piglets, Campylobacter prevalence was 7 of 15 on day 1 and 15 of 15 (100%) on day 20. These data suggest that successful permanent colonization of the gut by Campylobacter is probably related to constant exposure of piglets to Campylobacter-positive feces. Campylobacter prevalence may be diminished in neonates that are reared off-sow in isolated nurseries. PMID- 11041147 TI - Comparison of the attachment of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, and Pseudomonas fluorescens to lettuce leaves. AB - Attachment of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella Typhimurium, and Pseudomonas fluorescens on iceberg lettuce was evaluated by plate count and confocal scanning laser microscopy (CSLM). Attachment of each microorganism (approximately 10(8) CFU/ml) on the surface and the cut edge of lettuce leaves was determined. E. coli O157:H7 and L. monocytogenes attached preferentially to cut edges, while P. fluorescens attached preferentially to the intact surfaces. Differences in attachment at the two sites were greatest with L. monocytogenes. Salmonella Typhimurium attached equally to the two sites. At the surface, P. fluorescens attached in greatest number, followed by E. coli O157:H7, L. monocytogenes, and Salmonella Typhimurium. Attached microorganisms on lettuce were stained with fluorescein isothiocyanate and visualized by CSLM. Images at the surface and the cut edge of lettuce confirmed the plate count data. In addition, microcolony formation by P. fluorescens was observed on the lettuce surface. Some cells of each microorganism at the cut edge were located within the lettuce tissues, indicating that penetration occurred from the cut edge surface. The results of this study indicate that different species of microorganisms attach differently to lettuce structures, and CSLM can be successfully used to detect these differences. PMID- 11041148 TI - Listeria monocytogenes contamination pattern in pig slaughterhouses. AB - Ten low-capacity slaughterhouses were examined for Listeria by collecting a total of 373 samples, of which 50, 250, and 73 were taken from carcasses, pluck sets, and the slaughterhouse environment, respectively. Six slaughterhouses and 9% of all samples were positive for Listeria monocytogenes. Of the samples taken from pluck sets, 9% were positive for L. monocytogenes, the highest prevalence occurring in tongue and tonsil samples, at 14% and 12%, respectively. Six of 50 (12%) carcasses were contaminated with L. monocytogenes. In the slaughterhouse environment, L. monocytogenes was detected in two, one, one, and one sample originating from the saws, drain, door, and table, respectively. Carcasses were contaminated with L. monocytogenes in those two slaughterhouses, where the mechanical saws, used for both brisket and back splitting, were also positive for L. monocytogenes. A total of 58 L. monocytogenes isolates were characterized by pulsed-field gel electrophoresis typing. The isolates were divided into 18 pulsotypes, 15 of which were detected in pluck sets. In two slaughterhouses, where the carcasses were contaminated with L. monocytogenes, the same pulsotypes were also recovered from splitting saws. In addition, identical pulsotypes were recovered from pluck sets. Our findings indicate that L. monocytogenes of tongue and tonsil origin may contaminate the slaughtering equipment that may in turn spread the pathogen to carcasses. Thus, it is of the utmost importance to follow good manufacturing practices and to have efficient cleaning and disinfection procedures to prevent equipment being contaminated with L. monocytogenes. PMID- 11041149 TI - Improved detection of nontyphoid and typhoid Salmonellae with balanced agar formulations. AB - A strategically balanced medium was developed for the improved detection of nontyphoid and typhoid salmonellae. Its balanced sugar (cellobiose, lactose, mannitol, and trehalose) and protein (beef extract and polypeptone peptone) formulation provided Salmonella with a selective growth advantage over non Salmonella enteric organisms. The formulations promoted the production and detection of H2S production levels that otherwise might be missed with traditional agar formulations. In combination, these advantages contributed to increased sensitivity without the loss of specificity. In comparative studies using 86 samples of meat products (beef, pork, and chicken), the new media, Miller-Mallinson (MM) agar and xylose lysine tergitol (Niaproof) 4 agar, possessed significantly higher sensitivity (P < 0.001) and an improved specificity over bismuth sulfite, hektoen enteric, and xylose lysine desoxycholate agars. However, these samples did not contain nontyphoid salmonellae with weak to ultraweak H2S production characteristics. Modified formulations of MM agar were generally similar to bismuth sulfite and hektoen enteric agars in the identification of four of seven globally diverse strains of Salmonella serotype Typhi. Two of these seven strains were found to produce more readily identifiable black (H2S-positive) colonies on MM agar, whereas one of the seven was not readily detected by any of the media. The improved detection of nontyphoid and typhoid salmonellae attests to the sensitivity of MM agar and to its potentially broad utility in both clinical and food quality laboratories. PMID- 11041150 TI - Comparison of different peptidase substrates for evaluation of microbial quality of aerobically stored meats. AB - Different aminopeptidase and endopeptidase substrates were assessed for the detection of enzymatic activity of microorganisms collected from the surface of aerobically cold-stored pork and beef. The most sensitive substrates were fluorogenic Ala-7-amino-4-methylcoumarin (Ala-AMC) or Leu-AMC and colorogenic Ala p-nitroanilide (Ala-pNA). Activity on natural oligopeptides, e.g., bradykinin or alpha(s1) casein fragment 1 to 23, was very low. The correlation coefficient (r) between log surface counts of 66 meat samples and log fluorescence or absorbance after incubation of surface microbial cells for 2 h with Ala-AMC, Leu-AMC, and Ala-pNA was 0.89, 0.83, and 0.82, respectively. A distinct yellow color was obtained with Ala-pNA when the surface count was approximately 10(6) CFU/cm2. Although correlation and sensitivity was better, no clear advantage is obtained with the use of the fluorogenic Ala-AMC or Leu-AMC instead of Ala-pNA, a substrate proposed by Alvarado et al. (J. Food Sci. 57:1330, 1992) for rapidly assessing the microbial quality of refrigerated meat. The correlation coefficient (r) between time of cold storage and surface count was 0.69. PMID- 11041151 TI - Anatomy, biochemistry, and physiology of articular cartilage. AB - Articular cartilage serves as a load-bearing elastic material that is responsible for the frictionless movement of the surfaces of articulating joints. Its ability to undergo reversible deformation depends on its structural organization, including the specific arrangement of the matrix macromolecules and the chondrocytes. Interactions between the matrix and chondrocytes are responsible for the biological and mechanical properties of articular cartilage and enable it to respond by effecting a balance between anabolism and catabolism as well as continual internal remodeling. Age-related changes in the function of chondrocytes may contribute to the initiation and progression of osteoarthritis. PMID- 11041152 TI - Subchondral bone and cartilage disease: a rediscovered functional unit. AB - The role of subchondral bone in the pathogenesis of cartilage damage has likely been underestimated. Subchondral bone is not only an important shock absorber, but it may also be important for cartilage metabolism. Contrary to many drawings and published reports, the subchondral region is highly vascularized and vulnerable. Its terminal vessels have, in part, direct contact with the deepest hyaline cartilage layer. The perfusion of these vessels accounts for more than 50% of the glucose, oxygen, and water requirements of cartilage. Bony structure, local metabolism, hemodynamics, and vascularization of the subchondral region differ within a single joint and from one joint to another. Owing to these differences, repetitive, chronic overloading or perfusion abnormalities may result in no pathological reaction at all in one joint, while in another joint, these same conditions may lead to osteonecrosis, osteochondritis dissecans, or degenerative changes. According to this common etiological root, similar pathological reactions beginning with marrow edema and necrosis and followed by bone and cartilage fractures, joint deformity, and insufficient healing processes are found in osteonecrosis, osteochondritis dissecans, and degenerative disease as well. PMID- 11041153 TI - Physicochemical properties of normal articular cartilage and its MR appearance. AB - Basic physical and physicochemical properties of articular cartilage are correlated with the MR parameters of this tissue. From these parameters, the typical appearance of cartilage in MR images is deduced. Some practical implications for clinical utilization of MRI of articular cartilage are summarized. PMID- 11041154 TI - Magnetic resonance imaging of articular cartilage and evaluation of cartilage disease. AB - Clinical magnetic resonance imaging of articular cartilage is possible by using techniques that offer high contrast between articular cartilage and adjacent structures in reasonable examination times. The fat-suppressed, three dimensional, spoiled gradient-echo sequence has been reported to be accurate and reliable, and the addition of this sequence to a routine examination does not significantly compromise patient throughput. Fast spin-echo imaging also shows promise in the clinical evaluation of articular cartilage, because the newer, stronger-gradient systems allow thinner slice acquisition with two-dimensional sequences. Together, these sequences allow the evaluation of intrachondral lesions and surface defects. Furthermore, quantitative measurements of cartilage volume for follow-up studies are possible with the use of the fat-suppressed, three-dimensional, spoiled gradient-echo sequence. PMID- 11041155 TI - Magic-angle effect in magnetic resonance imaging of articular cartilage: a review. AB - RATIONALE AND OBJECTIVES: The laminar appearance of articular cartilage in magnetic resonance (MR) images has been a source of confusion, especially concerning the number, intensity, thickness, and origin of the layers. The laminar appearance is associated with the magic-angle effect in the MR imaging (MRI) of articular cartilage. METHODS: This article introduces the topic with background information about cartilage and the magic-angle effect and then reviews the literature about the magic-angle effect. The review concludes with a brief discussion of the future directions of study and the potential clinical relevance of the laminae in MR images of articular cartilage. CONCLUSIONS: The magic-angle effect is commonly seen in MR images of several tissues. The direct cause of the laminar appearance of articular cartilage is the T2 relaxation anisotropy in the tissue, which is closely linked to the structure of the collagen fibers, their orientation in the magnetic field, and the water proteoglycan interaction that amplifies the prevailing orientation of the collagen fiber network. The laminar appearance of cartilage has an intrinsic spatial heterogeneity over the two-dimensional joint surface, which leads to inconsistencies in the reported total number of cartilage laminae and the laminar patterns observable in MRI, depending on where the sample was taken. Two additional thin, low-intensity laminae may also be visible at the boundaries of the cartilage with fluid and with bone; whether these boundary laminae are identified and counted with the others may introduce inconsistency in the results reported by various researchers. PMID- 11041156 TI - MRI techniques in early stages of cartilage disease. AB - Cartilage degenerative diseases affect millions of people. Our understanding of these diseases and our ability to establish efficacious treatment strategies have been confounded by the difficulty of nondestructively evaluating the state of cartilage. Imaging strategies that allow visualization of cartilage integrity would revolutionize the field by allowing us to visualize early stages of degeneration and thus to evaluate predisposing factors for cartilage disease and changes resulting from interventions (eg, therapies) in culture studies, tissue engineered systems, animal models, and in vivo in humans. Here we briefly review current state-of-the-art MRI strategies relevant to understanding and following treatment in early cartilage degeneration. We review MRI as applied to the assessment of the whole joint, of cartilage as a whole (as an organ), of cartilage tissue, and of cartilage molecular composition and structure. Each of these levels is amenable to assessment by MRI and offers different information that, in the long run, will serve as an important element of cartilage imaging. PMID- 11041157 TI - Treatment of articular cartilage defects. PMID- 11041158 TI - Increased potassium concentration inhibits stimulation of vascular smooth muscle proliferation by PDGF-BB and bFGF. AB - The effects of changes in extracellular potassium concentration on the rate of vascular smooth muscle cell proliferation stimulated by cytokines and serum were analyzed in vitro. To analyze the DNA synthesis response, cells from swine coronary artery were grown in DMEM medium containing 3, 4, 5, or 6 mmol/L potassium together with 20 ng/mL platelet-derived growth factor BB (PDGF-BB), 25 ng/mL basic fibroblast growth factor (bFGF), or 5% fetal bovine serum (FBS), with [methyl 3H] thymidine added, for 24 h. Proliferation responses were analyzed in cells grown in medium with potassium concentrations of 3, 4, 5, or 6 mmol/L, together with either 20 ng/mL PDGF-BB, 25 ng/mL bFGF, or 5% FBS, for 7 days, then harvested and counted. Highly significant inverse relationships were observed between potassium concentration and 3H-thymidine incorporation stimulated by each of the three agonists (P < .01 for each, ANOVA), and between potassium concentration and proliferation (all P < .01, ANOVA). Elevation of potassium concentration within the physiologic range inhibits vascular smooth muscle cell DNA synthesis and proliferation. PMID- 11041159 TI - Treatment of hypertensive children with amlodipine. AB - Amlodipine, a long-acting dihydropyridine calcium channel blocking agent, was administered to 55 children (age: 11.5 +/- 5.4 years) with hypertension, 49 of whom (89%) had secondary hypertension. Efficacy was assessed by comparing pretreatment blood pressure (BP) to follow-up BP obtained in our outpatient Pediatric Nephrology clinic. Thirty-two (58%) patients achieved BP control with amlodipine alone, and 31 (55%) patients received amlodipine twice daily. Eleven patients received amlodipine as a suspension. Mean amlodipine dose was 0.16 +/- 0.12 mg/kg/day; there was an inverse relationship between patient age and amlodipine dose. Follow-up BP were significantly lower than pretreatment BP: systolic BP fell from 129 +/- 12 to 122 +/- 12 mm Hg (P = .004), and diastolic BP fell from 78 +/- 13 to 70 +/- 19 mm Hg (P = .003). A small, clinically insignificant increase in heart rate (from 91 +/- 19 beats/min to 99 +/- 26 beats/min; P = .02) occurred during amlodipine treatment. Adverse effects reported included dizziness (three patients), fatigue (two patients), flushing (two patients), and leg edema (one patient). All improved with dose reduction. We conclude that amlodipine provides effective BP control without significant adverse effects in children with hypertension, and can be used as monotherapy in most children. Young children appear to require significantly higher doses per kilogram of body weight than older children. Twice-daily dosing may be required in many children to achieve BP control. Detailed pharmacokinetic studies are needed to confirm these observations. PMID- 11041160 TI - The influence of physical activity on the variability of ambulatory blood pressure. AB - The aim of this study was to assess the contribution of physical activity levels to blood pressure (BP) variability, and to assess the effect age, gender, body mass index, and use of antihypertensive medications on this relationship. We simultaneously monitored 24-h ambulatory BP by automated recorder and activity by actigraphy in 431 patients. Mean activity scores for the 5, 10, 15, and 20 min preceding each BP measurement were calculated, and BP and heart rate were related to these variables using linear mixed model regression. Various patient characteristics were added to the mixed model as covariates. Patients were heterogeneous in age (48 +/- 13 years), sex (49% men), and average 24-h BP (132/81 +/- 15/10 mm Hg). Mean daytime activity level was 44 +/- 15 U. During the daytime, systolic BP (r = 0.33), diastolic BP (r = 0.29), and heart rate (r = 0.42) correlated best with the average activity for the 15 min preceding each measurement (P < .001). Variance was very high, with activity explaining from 0% to 62% of BP variability for different individuals. Men and the obese had a greater reactivity of systolic BP to activity; older patients and those on antihypertensive therapy had a lower reactivity of heart rate. Blood pressure level is significantly associated with physical activity, but the percentage of variance of BP explained by physical activity varies greatly between individuals. Correlation is strongest between BP and average activity integrated over the previous 15 min. Much of the variance in blood pressure remains unexplained. PMID- 11041161 TI - Changes in cardiac energy metabolism during early development of female SHR. AB - We investigated effects of hypertension and early development on myocardial energy metabolism as reflected by maximal enzyme activities, glucose transporter content, and endogenous substrates in female Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). Left ventricular hypertrophy and systolic hypertension were evident in SHR at 6 weeks of age and these differences increased at 14 and 22 weeks of age. 3-Hydroxyacyl-CoA dehydrogenase (HOAD) activity in the left ventricle was 18% lower in 6-week-old rats than both 14- and 22-week-old rats, but not different between WKY rats and SHR. Hexokinase activity was 15% lower in 6-week-old SHR than WKY rats and decreased progressively with age in both strains. Glucose transporter (GLUT) 1 content was nearly twofold greater in 6-week-old rats than both 14- and 22-week-old rats. We found no difference in citrate synthase activity or GLUT4 content among groups. Glycogen concentration was 44% lower in SHR than WKY rats, whereas triglyceride was slightly (16%) higher in SHR than WKY rats. Older animals had higher levels both glycogen and triglyceride than younger animals. We conclude that the left ventricle of both SHR and WKY rats may change from predominantly glucose to fatty acid oxidation for energy production during early development. PMID- 11041162 TI - Heart rate as a predictor of future blood pressure in schoolchildren. AB - Heart rate (HR) has been shown to predict future blood pressures (BP) in studies in adults. We explored the relation of HR to future BP levels in a cohort of 344 black and 456 white schoolchildren ages 5 to 19 years, to examine the hypothesis that HR predicts subsequent BP even very early in life. After making baseline measurements, BP was assessed longitudinally 1 to 24 additional times (mean = 8.25) after the baseline period, at intervals of approximately 6 months. We found that HR was significantly related to future diastolic BP in the black boys (P = .016) after adjusting for baseline diastolic BP, age, and body mass index, but not in the black girls or in the white children. Because HR is reflective of sympathetic nervous system (SNS) activity that in turn can be related to the renin-angiotensin system (RAS), we also explored the relation of HR to the RAS by studying relationships to variants in the angiotensinogen gene and the angiotensin I-converting enzyme (ACE) gene. We found a significantly positive relationship of HR to the presence of the deletion allele of the ACE gene (P = .0015), but, again, only in the black boys. Because blacks in general appear to retain additional sodium when compared with whites, the SNS, as reflected in the HR, may influence BP more when individuals have increased sodium retention. In summary, baseline HR predicted future diastolic BP in the black boys but not in the black girls or in the white children. PMID- 11041163 TI - Effects of imidapril on endothelin-1 and ACE gene expression in failing hearts of salt-sensitive hypertensive rats. AB - The renin-angiotensin system and endothelin are important regulators of the cardiovascular system. Although increased production of endothelin-1 (ET-1) is reported in patients with heart failure, the detailed mechanism remains to be determined. To elucidate the relationship between the renin-angiotensin system and ET-1 in hypertensive heart failure, we evaluated the effects of long-term treatment with imidapril, an angiotensin converting enzyme (ACE) inhibitor, on preproET-1, endothelin A receptor (ETAR), and ACE mRNA expression in the left ventricle and evaluated these in relation to myocardial remodeling in the failing heart of Dahl salt-sensitive (DS) hypertensive rats fed a high salt diet. In DS rats fed an 8% NaCl diet after the age of 6 weeks, a stage of concentric left ventricular hypertrophy at 11 weeks (DSLVH) was followed by a distinct stage of left ventricular failure with chamber dilatation at 18 weeks (DSHF). Imidapril (DSHF-IM, n = 8, 1 mg/kg/day, subdepressor dose) or vehicle (DSHF-V, n = 8) was given from stage DSLVH to DSHF for 7 weeks, and age-matched (18 weeks) Dahl salt resistant rats fed the same diet served as the control group (DR-C, n = 8). In both groups, blood pressure was similar and significantly higher than in DR-C. Markedly increased left ventricular end-diastolic diameter and reduced fractional shortening in DSHF-V was significantly ameliorated in DSHF-IM using transthoracic echocardiography. The preproET-1, ETAR, and ACE mRNA levels in the left ventricle were significantly increased in DSHF-V compared with DR-C, and significantly suppressed in DSHF-IM compared with DSHF-V. DSHF-V demonstrated a significant increase in the wall-to-lumen ratio and perivascular fibrosis in coronary arterioles, and myocardial fibrosis, with all these parameters being significantly improved by imidapril. In conclusion, myocardial remodeling and heart failure in DS rats fed a high salt diet were significantly ameliorated by a subdepressor dose of imidapril, which may be attributable to a decrease in ET-1 mRNA expression and angiotensin II in the left ventricle. PMID- 11041165 TI - Left ventricular mass and atrial volume determined by cine magnetic resonance imaging in essential hypertension. AB - To evaluate the relationship between left atrial volume determined by cine magnetic resonance imaging and progression of left ventricular hypertrophy (LVH), left atrial volume and echocardiographic left ventricular mass (LVM) were measured in 30 hypertensive patients (15 without LVH and 15 with LVH) and 10 normotensive control subjects. We also evaluated the effects of antihypertensive therapy on the cardiac chamber volumes and LVM in hypertensive patients. The cardiac chamber volumes and LVM were indexed by body surface area. Although there were no significant differences in left ventricular chamber volumes among the three groups, both maximum and minimum left atrial volume indexes, and the LVM index were greater in hypertensive patients with LVH than in the other two groups. The LVM index was correlated with maximum left atrial volume index (r = 0.74, P < .0001), and minimum left atrial volume index (r = 0.76, P < .0001), respectively. Furthermore, in multivariate models, the LVM index was significantly correlated with maximum left atrial volume index. In hypertensive patients with LVH, both maximum and minimum left atrial volume indexes, and the LVM index significantly reduced after treatment. The percent of changes in maximum left atrial volume index after treatment was significantly correlated with the percent of changes in LVM index after treatment. In conclusion, our data indicate that LVH is an independent determinant of left atrial enlargement, and both LVH and left atrial enlargement may be reversed by some effective therapeutic interventions. PMID- 11041164 TI - Dexamethasone worsens nitric oxide inhibition-induced hypertension and renal dysfunction. AB - Chronic nitric oxide (NO) inhibition with Nomega-nitro-L-arginine methyl ester (L NAME) has previously been reported to produce systemic hypertension, renal vasoconstriction, and renal damage. In this study we investigated whether a compensatory restoration of NO synthesis occurs in chronic L-NAME hypertension and whether chronic treatment with dexamethasone (Dex) (which inhibits inducible NO synthase [iNOS]) can influence the course of the hypertension. We found that in the conscious chronically L-NAME-treated (approximately =10 mg/kg/24 h) hypertensive rats, acute systemic NOS inhibition elicited a further increase in blood pressure (BP), indicating partial restoration of NO production. Chronic Dex in a dose previously reported to inhibit iNOS (5 microg/24 h), amplified the hypertension (within 2 days), renal vasoconstriction, and reduction in glomerular filtration rate because of L-NAME. In contrast, chronic Dex alone had no effects on renal hemodynamics or BP during the first week, although by the end of week 2 a small increase in BP (approximately =10 mm Hg) was evident. These results show that BP continues to increase with chronic L-NAME despite partial restoration of NO production. An iNOS, which might be stimulated and escaped inhibition by L NAME, may be responsible for the compensatory restoration of NO synthesis, serving to attenuate the development of hypertension and renal dysfunction. PMID- 11041166 TI - Antihypertensive and antihypertrophic effects of omapatrilat in SHR. AB - Vasopeptidase inhibitors, such as omapatrilat are single molecules that simultaneously inhibit neutral endopeptidase (NEP) and angiotensin converting enzyme (ACE). In normotensive rats, a single dose of oral omapatrilat (10 mg/kg) and 1 mg/kg inhibited plasma ACE (P < .01) for 24 h and increased plasma renin activity for 8 h (P < .01). In vitro autoradiography using the specific NEP inhibitor radioligand 125I-RB104 and the specific ACE inhibitor radioligand 125I MK351A showed omapatrilat (10 mg/kg) caused rapid and potent inhibition of renal NEP and ACE, respectively, for 24 h (P < .01). In spontaneously hypertensive rats, 10 days of oral omapatrilat (40 mg/kg/day) reduced blood pressure (vehicle 237 +/- 4 mm Hg; omapatrilat, 10 mg/kg, 212 +/- 4 mm Hg; omapatrilat 40 mg/kg, 197 +/- 4 mm Hg, P < .01) in a dose-dependent manner (10 v 40 mg/kg, P < .01). Left ventricular hypertrophy was significantly reduced by high-dose omapatrilat (vehicle 2.76 +/- 0.03 mg/g body weight; omapatrilat, 10 mg/kg, 2.71 +/- 0.02 mg/g; omapatrilat 40 mg/kg, 2.55 +/- 0.02 mg/g, P < .01) and omapatrilat also increased kidney weight compared to vehicle (both doses, P < .01). Omapatrilat caused significant inhibition of plasma ACE and increased plasma renin activity (both doses, P < .01), and in vitro autoradiographic studies indicated sustained inhibition of renal ACE and NEP (both doses, P < .01). Omapatrilat is a potent vasopeptidase inhibitor, and its antihypertensive effects are associated with inhibition of NEP and ACE at the tissue level and beneficial effects on cardiovascular structure. Relating the degree of tissue inhibition to physiologic responses may allow further definition of the role of local renin angiotensin and natriuretic peptide systems in the beneficial effects of vasopeptidase inhibitors. PMID- 11041167 TI - Angiotensin II regulates the cell cycle of vascular smooth muscle cells from SHR. AB - We have demonstrated that spontaneously hypertensive rats (SHR)-derived vascular smooth muscle cells (VSMC) show the exaggerated growth and produce angiotensin II (Ang II). In the current study, we investigated the role of endogenous Ang II in the regulation of the cell cycle in VSMC from SHR. Levels of Ang II in conditioned medium from SHR-derived VSMC cultured without serum were significantly higher than levels in conditioned medium from Wistar-Kyoto (WKY) rat-derived VSMC. Basal DNA synthesis was higher in quiescent VSMC from SHR than that in cells from WKY rats. An Ang II type 1 receptor antagonist, CV11974, significantly inhibited the elevation in DNA synthesis in quiescent VSMC from SHR but did not affect it in cells from WKY rats. Cellular DNA content analysis by flow cytometry revealed that the proportion of cells in S phase was higher, whereas the proportion of cells in G1+G0 phase was lower in VSMC from SHR than those in cells from WKY rats. CV11974 significantly decreased the proportion of cells in S phase and correspondingly increased the proportion of cells in G1+G0 phase in VSMC from SHR, but it did not affect the proportion in cells from WKY rats. Cyclin-dependent kinase 2 (CDK2) activity, which is known to induce the progression from G1 to S phase, was higher in VSMC from SHR than in cells from WKY rats. Expression of CDK2 inhibitor p27(kip1) mRNA was markedly higher in VSMC from SHR than in cells from WKY rats. CV11974 decreased expression of p27(kip1) mRNA in VSMC from SHR, whereas CV11974 increased it in cells from WKY rats. These findings indicate that enhanced production of endogenous Ang II regulates the cell cycle especially in the progression from G1 to S phase, and increases CDK2 activity, which is independent of p27(kip1) in VSMC from SHR. PMID- 11041168 TI - Nitric oxide is an excitatory modulator in the rostral ventrolateral medulla in rats. AB - Nitric oxide is a messenger molecule having various functions in the brain. Previous studies have reported conflicting results for the roles of nitric oxide in the rostral ventrolateral medulla, a major center that regulates sympathetic and cardiovascular activities. We hypothesized that in this region, nitric oxide may have a biphasic effect on cardiovascular activity. Microinjection of a low dose (1 nmol) of a nitric oxide donor sodium nitroprusside or a cyclic GMP agonist 8-bromocyclic GMP into this area increased arterial pressure, whereas injection of a nitric oxide synthase inhibitor Nomega-nitro-L-arginine methyl ester or a soluble guanylate cyclase inhibitor methylene blue decreased arterial pressure. Microinjection of a high dose (100 nmol) of sodium nitroprusside decreased arterial pressure and inhibited spontaneous respiration with concomitant production of peroxynitrite, a strong cytotoxic oxidant. Increases in arterial pressure caused by microinjection of L-glutamate were inhibited after preinjection of Nomega-nitro-L-arginine methyl ester or methylene blue. Increases in arterial pressure caused by microinjection of sodium nitroprusside (1 nmol) were inhibited after preinjection of a glutamate receptor antagonist kynurenate. These results suggest that low doses of nitric oxide may increase arterial pressure, whereas high doses of nitric oxide may decrease arterial pressure through cytotoxic effects in the rostral ventrolateral medulla. They also indicate that nitric oxide may stimulate neurons both through activation of the nitric oxide cyclic GMP pathway and through modulation of glutamate receptor stimulation, and therefore, increase arterial pressure in rats. PMID- 11041169 TI - White coat effect of alcohol. AB - Numerous studies have shown a relationship between alcohol intake and elevated clinic blood pressures (BP). However, there have been few studies on the relationship between alcohol consumption and 24-h ambulatory BP monitoring. This study aimed to determine the relationship between alcohol intake, clinic BP, and 24-h ambulatory BP recordings to determine to what extent a white coat effect may contribute to the relationship between alcohol consumption and BP. Clinical BP and 24-h ambulatory BP were measured in 121 male volunteers aged 50.6 +/- 9.8 years (range, 30-70 years) who consumed between 0 and 2050 g of alcohol per week (mean, 394 +/- 342 g; median, 385 g/week). Supine clinical systolic BP (SBP) was significantly related to alcohol intake (beta = 0.242; P = .007). Alcohol consumption was not related to 24-h mean SBP or diastolic BP (DBP), daytime SBP or DBP, or nighttime SBP or DBP (daytime SBP: beta = 0.02, P = .802). Alcohol intake was significantly related to the difference between clinic SBP and mean daytime SBP (beta = 0.260, P = .004). Twenty-four-hour mean heart rate (HR), daytime mean and nighttime mean HR were strongly associated with alcohol intake (24-h HR: beta = 0.455, P < .001). These results suggest that the association between alcohol consumption and elevated BP is contributed to by a significant white coat effect and that excessive alcohol consumption may be a significant factor in explaining differences between clinic and ambulatory BP measurements. PMID- 11041170 TI - Etiology and pathophysiology of stroke as a complex trait. AB - Stroke (brain attack) is currently the third leading cause of death in Western societies. Recent advances in molecular genetics have finally demonstrated what has long been suggested by the clinical observation, that is, stroke is not only the complication of major pathologic conditions such as atherosclerosis, hypertension, or cardiac diseases, but rather it represents a complex trait itself. Thus, the pathogenesis of stroke is often the result of the combined effects of genes exerting a direct contributory role and of their interactions with several environmental determinants. A genetic dissection of stroke has been attempted in suitable animal models and in humans. With this approach, the genetic defects underlying monogenic disorders associated with stroke were identified. Moreover, important findings have recently highlighted the contribution of genes encoding cardiovascular hormones, such as the atrial natriuretic peptide, for the pathogenesis of multifactorial, polygenic forms of stroke. A more thorough understanding of the fine mechanisms, dependent from mutations within stroke susceptibility genes and underlying the disease pathogenesis, may help to introduce new specific tools to achieve better prevention and treatment of stroke. PMID- 11041171 TI - The pathophysiology of mitral regurgitation. AB - The results of treatment of heart valve disease have improved steadily during the past 20 years. In aortic stenosis, although postoperative survival rates approximated those of age-matched controls, the outcome of surgery to treat ischemic and non-ischemic mitral regurgitation was grave. The reasons for this were two-fold: first, patients were referred for surgery late in the course of their disease, when irreversible left ventricular (LV) dysfunction prevented postoperative restoration of contractile function. Second, the value of the mitral valve apparatus in facilitating LV contraction was unrecognized, and this structure was often removed at surgery, in turn worsening pre-existent LV dysfunction. Consequently, patients with LV dysfunction due to mitral regurgitation underwent surgery that caused further damage to the left ventricle. Not surprisingly, postoperative LV function was poor, congestive heart failure persistent, and lifespan shortened. More recently, however, substantial insight has been gained into the value of the mitral valve apparatus, the causes of LV dysfunction in mitral regurgitation, and into the objective markers of LV function that permit the clinician to recommend surgery before muscle dysfunction has become severe and irreversible. PMID- 11041172 TI - Effect of Inoue balloon mitral valvotomy on severe pulmonary arterial hypertension in 315 patients with rheumatic mitral stenosis: immediate and long term results. AB - BACKGROUND AND AIMS OF THE STUDY: Despite advances in surgical techniques, mitral valve surgery in patients with severe pulmonary arterial hypertension (PAH) causes considerable mortality and morbidity. Balloon mitral valvotomy (BMV) is an established alternative to treat high-risk surgical patients with mitral stenosis (MS). The study aims were to evaluate immediate and long-term efficacy of BMV in patients with MS and severe PAH, compared to those with mild/moderate PAH. METHODS: Among 1,125 patients who underwent Inoue BMV, 315 had severe PAH (mean pulmonary artery (PA) pressure > or = 50 mmHg (group I; 79 of these patients had suprasystemic PAH). Results from this group were compared with those of patients with mild/moderate PAH (group II). RESULTS: Group I patients were younger and more symptomatic (mean PA pressure 62 +/- 10.6 mmHg versus 32.6 +/- 8.2 mmHg in group II). Before BMV, mean transmitral gradient (17.8 +/- 6.5 versus 14.4 +/- 5.4 mmHg) and pulmonary capillary wedge pressure (PCWP) (31.6 +/- 6.1 versus 22.8 +/- 6.2 mmHg) were significantly higher, while mitral valve area (MVA) (0.66 +/- 0.2 versus 0.85 +/- 0.2 cm2) was significantly lower in group I. After BMV, PA mean pressure was significantly reduced (34.8 +/- 11.2 and 21.1 +/- 8.4 mmHg), transmitral gradient (8.0 +/- 3.9 and 6.9 +/- 3.2 mmHg) and mean PCWP (12.8 +/- 5.8 and 11.0 +/- 5.1 mmHg) in groups I and II, respectively, with a comparable increase in MVA (1.77 +/- 0.4 and 1.84 +/- 0.5 cm2). Group I patients had worse baseline hemodynamic parameters than group II, but the former had a higher absolute gain in hemodynamic parameters. Residual severe PAH after BMV was seen in 9.8% of patients, with PA pressures normalized in 9.5%. Among 79 patients with suprasystemic PA pressure (mean PA systolic pressure 116.6 +/- 28.2 mmHg), 16.5% normalized their PA pressures and 25.3% had residual severe PAH. At mean follow up of 33 months, 80.4% were in NYHA class I. Mean PA systolic pressure in 161 patients was 39.0 +/- 14.2 mmHg compared with a post-BMV value of 55.0 +/- 16.9 mmHg; thus, a sustained fall in pressure was demonstrated at follow up. CONCLUSION: Inoue BMV is safe and effective in patients with MS and severe PAH. Although these patients have worse clinical and hemodynamic parameters before BMV, they achieve a greater absolute gain in terms of improvement in all hemodynamic parameters. PMID- 11041173 TI - Superiority of mitral valve replacement with preservation of subvalvular structure to conventional replacement in severe rheumatic mitral valve disease: a modified technique and results of one-year follow up. AB - BACKGROUND AND AIM OF THE STUDY: Mitral valve replacement with preservation of the subvalvular apparatus (MRVP) has been proven superior to conventional mitral valve replacement (MVR). We devised a simple modified MVRP method in this prospective, randomized study to investigate the clinical effects and one-year follow up echocardiographic results of MVRP compared with MVR in patients with severe rheumatic mitral insufficiency (MI). METHODS: Sixty-eight patients with severe rheumatic MI with or without stenosis were randomized to MVRP (n = 35) and MVR (n = 33) groups. In MVRP patients, the preserved tissue was pulled back posteriorly to the posterior wall of the left ventricle, then plicated and reaffixed to one-fourth of the annular circumference in the posterior annulus, in order to prevent left ventricular outflow tract (LVOT) obstruction. Clinical data including cumulative ventricular arrhythmias and use of inotropes were collected. Echocardiography examination was performed before surgery, and at five days, three months and one year thereafter. RESULTS: There were no preoperative differences patient data. The cross-clamp time was 2.2 min longer in MVRP patients. The one-month mortality rate after surgery was lower in MVRP patients (2.9% versus 15.2%, p = 0.074). Mechanical ventilation and ICU times were shorter in the MVRP group (17.6 versus 24.8 and 52.5 versus 70.6 h, p = 0.001 and 0.1, respectively). There were fewer ventricular arrhythmias and less need for inotropic support in this group. One year follow up echocardiography data showed better preserved left ventricular ejection fraction (LVEF) and better recovery of heart size after MRVP. There was no indication that preserved valvular tissue interfered with mechanical valve function, or caused LVOT obstruction. CONCLUSION: This modified MVRP technique is simple, effective and without risk of LVOT obstruction. In severe rheumatic MI patients the outcome of MVRP is superior to that of conventional MVR in term's of mortality, postoperative care needs, left ventricular function and heart dimensions. PMID- 11041174 TI - Long-term results of balloon aortic valvulotomy for congenital aortic stenosis in children and adolescents. AB - BACKGROUND AND AIM OF THE STUDY: The aim of the study was to report on the long term results of aortic valve balloon dilatation (AVBD) for congenital valvular aortic stenosis in children and young adults. METHODS: The records of 74 patients (age range: 1-20 years) who underwent AVBD at a single center were analyzed retrospectively. Special attention was paid to factors that might influence long term outcome. RESULTS: The procedure was successful in 71 patients (96%). The mean (+/- SD) reduction in peak-to-peak systolic gradient (PSG) was 68.7 +/- 13.5%. No patient required immediate surgical intervention. Survival after dilatation was 100% at 12 years. At follow up (mean 5.5 +/- 2.9 years; range: 2 12 years), 20% of patients had restenosis and 21% had significant aortic regurgitation (AR) (grade > or = 3). Reintervention was performed in 14% of patients. Severity of AR and high residual stenosis immediately after AVBD were associated with the late event rates. The actuarial intervention-free rates at five, seven and 12 years were 92.9%, 84.4% and 60%, respectively. CONCLUSION: AVBD is a useful, albeit palliative, procedure for children and young adults with significant congenital valvular aortic stenosis. PMID- 11041175 TI - Late conduction defects following aortic valve replacement. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to determine the incidence and clinical significance of late cardiac conduction defects (CD) after aortic valve replacement (AVR). METHODS: An analysis was made of 100 consecutive cases after AVR in a prospective outpatient evaluation program. RESULTS: The perioperative (30-day) mortality rate was 5%, and incidence of perioperative pacemaker implantation 3%. Among patients, 19% had CDs before surgery; a normal ECG was present during all periods in 45% of patients. The most frequent perioperative CD was left anterior hemiblock (LAHB; n = 8), and the most frequent late CD was left bundle branch block (LBBB; n = 8). Overall, 13.7% of operative survivors with normal preoperative and perioperative ECGs developed late CDs; one patient (1%) required pacemaker implantation 82 months after AVR. A further three patients (3%) had worsening of pre-existent CDs. Late CDs occurred over a wide time range (3 to 102 months) after surgery. CONCLUSION: There is an important incidence of CDs that occur late after AVR, even if the perioperative ECGs are normal; however, a need for late pacemaker implantation is rare. As CDs may occur at any time after surgery, regular follow up with precise evaluation of ECGs is called for. PMID- 11041176 TI - Necrotizing granulomata of the aortic valve in Wegener's disease. AB - Wegener's disease is an inflammatory disease of unknown etiology, characterized by a granulomatous-necrotizing general vasculitis. Cardiac involvement in the form of aortic pathology is not frequent. We report a case of Wegener's granulomatosis which required prosthetic aortic valve replacement for aortic valve insufficiency. Microscopic examination of the valve demonstrated histopathology typical of Wegener's disease. PMID- 11041177 TI - Replacement of the tricuspid valve in children with congenital cardiac malformations. AB - BACKGROUND AND AIM OF THE STUDY: When replacing the regurgitant tricuspid valve in children, the decision to use either a bioprosthesis or a mechanical valve remains controversial. METHODS: The atrioventricular valve for the pulmonary circulation was replaced in 11 young patients aged between 8 months and 13 years. Complications of congenital cardiac malformation were present in seven patients, and Ebstein's anomaly in three; tricuspid valvular regurgitation was an isolated lesion in one patient. A bioprosthesis was implanted on seven occasions, and a bileaflet mechanical valve on eight, including re-replacement of the valve in four patients. RESULTS: One patient died two years after implantation due to respiratory problems. Tricuspid stenosis due to valve calcification occurred in four bioprostheses at between four and nine years after initial replacement (57%). In three of these cases the native valve leaflets had not been removed. Thrombosis occurred in one patient with a mechanical valve; rereplacement was successful. Anticoagulant-related hemorrhage occurred in another patient. Among patients receiving a mechanical valve, 83% of valves were dysfunction-free after five and ten years. CONCLUSION: When replacing an atrioventricular valve for the pulmonary circulation in children, we prefer to use a low-profile mechanical valve, especially when extensive repair of intracardiac malformation has been carried out, but ventricular function is good. In children with poor cardiac performance, a bioprosthesis is preferred, with total resection of the native valve leaflets. PMID- 11041178 TI - Edge-to-edge repair of congenital familiar tricuspid regurgitation: case report. AB - We report a case of edge-to-edge (Alfieri's technique) repair of congenital familiar tricuspid regurgitation in a 49-year-old woman, who had severe tricuspid regurgitation, atrial septal defect with left-to-right shunt, and two stenoses in peripheral branches of the left pulmonary artery, of no clinical relevance. The repair was performed through a longitudinal inferior partial sternotomy. The atrial septal defect was closed by direct suture; the anterior and posterior leaflets of the tricuspid valve were sutured together. The chordae to the prolapsing medial part of the anterior leaflet were shortened by direct suture to the leaflet free edge. Annuloplasty was performed by means of a Carpentier ring. The final step was edge-to-edge approximation of the septal leaflet to the new antero-posterior position with two interrupted stitches. The hemodynamic result was excellent, and the patient eventually returned to full active life. PMID- 11041179 TI - Clinical and hemodynamic performance of the Toronto SPV bioprosthesis. AB - BACKGROUND AND AIM OF THE STUDY: By providing a superior hemodynamic profile, the stentless valve design allows ventricular remodeling and may improve patient survival after aortic valve replacement (AVR). Compared with stent-mounted prostheses, implantation is more complex and requires a longer ischemic time; this may adversely affect surgical risk, especially if patients are elderly or require a concomitant procedure. The mid-term clinical and hemodynamic performance of the Toronto SPV bioprosthesis in a predominantly elderly patient group was analyzed. METHODS: A total of 123 patients (median age 72 years) underwent AVR with the Toronto SPV. Concomitant procedures (mainly coronary artery bypass grafting, CABG), were performed in 60 patients (49%). Clinical details were recorded, with 100% follow up (total 317 patient-years). Hemodynamic evaluation, by serial echocardiography, was performed at four and 18 months after implantation. RESULTS: The early mortality rate was low (0.8%). Mean (+/- SD) actuarial survival at 53 months was 78 +/- 5.9%, with most patients (91%) in NYHA classes I and II. Freedom from valve-related complications were: endocarditis 93.8 +/- 2.3%, thromboembolism 90.3 +/- 3.7% and bleeding 95.8 +/- 1.8%; there were no structural failures. The valve hemodynamic profile was excellent for all sizes: peak gradient 8.8 +/- 4.3 mmHg, effective orifice area 1.9 +/- 0.54 cm2 with significant improvement in left ventricular fractional shortening. CONCLUSION: In this patient population the Toronto SPV was a suitable choice. Advanced age, a requirement for concomitant procedures and increased ischemic times did not adversely affect surgical risk. AVR with the Toronto SPV provided an excellent hemodynamic profile, and improved both left ventricular function and NYHA functional class. PMID- 11041180 TI - The Mosaic bioprosthesis in the aortic position at five years. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to collect intermediate-term data on the Mosaic bioprosthesis implanted in the aortic position. The device has been in clinical use since February 1994. METHODS: The Mosaic bioprosthesis is a stented porcine aortic valve, which combines a zero pressure differential fixation technique and anti-mineralization treatment with amino oleic acid for improved tissue durability. Between February 1994 and May 1999, 100 patients (49 females, 51 males; mean age at implant 73.4 +/- 7.3 years (range: 31-87 years) underwent aortic valve replacement with the Mosaic prosthesis in our department. Concomitant procedures were performed in 40% of cases. Patients were followed up prospectively at annual intervals; the mean follow up was 2.7 years (total 273.7 patient-years (pt-yr)) and was 100% complete. RESULTS: Total early mortality (within 30 days) was 3.0%; the late mortality rate was 4.4%/pt-yr and included a valve-related mortality rate of 0.7%/pt-yr. The freedom from event rates at five years were 97.3 +/- 1.9% for permanent neurological, 99.0 +/- 1.0% for transient neurological, 95.9 +/- 3.2 for thrombosed prosthesis, 95.6 +/- 2.2% for anti thromboembolic-related hemorrhage, 100% for primary valvular leak, 96.9 +/- 3.0% for non-structural dysfunction, 100% for endocarditis, and 92.0 +/- 4.9% for explant. The mean systolic gradients were 15.2, 13.1 and 10.1 mmHg for the 21, 23 and 25 mm valve sizes, respectively. CONCLUSION: The clinical and hemodynamic performance of the Mosaic prosthesis was highly satisfactory during the first five years after clinical introduction. Further data will be necessary to confirm long-term durability. PMID- 11041181 TI - An echocardiographic description of the Sulzer Carbomedics Synergy ST (Labcor) porcine valve in the aortic position. AB - BACKGROUND AND AIM OF THE STUDY: The Synergy ST (previously Labcor) valve is constructed from three separate porcine aortic cusps mounted on a scalloped, flexible, acetal copolymer stent. To date, no hemodynamic data describing this valve have been published. The aim of this study was to describe the echocardiographic findings in a population of unselected patients. METHODS: One investigator, who was blinded to valve size, studied 84 patients (31 females, 53 males; mean age 58 years; range: 19-83 years) at four centers in Brazil. The valves were studied at a mean of 31 months after implantation (range: 1-116 months). Mean gradient was calculated using the long modified Bernoulli equation, effective orifice area (EOA) using the classical from of the continuity equation, and resistance as mean gradient/flow. RESULTS: There were no significant differences in peak transaortic velocity and mean gradient between valves of different annulus size. However, EOA was directly related (p = 0.0075) and resistance indirectly related to valve size (p = 0.021, ANOVA). Trivial or mild regurgitation was seen in 31 patients (37%), moderate in three (4%) and severe in two (2%). The regurgitation was solely paraprosthetic in five patients, solely through the valve in 23, and both in seven. Regurgitation through the valve usually occurred adjacent to the commissures. CONCLUSION: The Synergy ST porcine valve has hemodynamics of forward flow similar to that of other stented porcine valves. PMID- 11041182 TI - Two years' clinical experience with a quadrileaflet stentless bioprosthesis in the mitral position. AB - BACKGROUND AND AIMS OF THE STUDY: Currently available bioprosthetic mitral valves do not provide sufficient durability. A new stentless pericardial prosthesis was designed for better hemodynamic performance and reduction of stress load compared with current stented bioprostheses. METHODS: Between September 1997 and August 1999, the Quadrileaflet mitral valve (QMV) was implanted in 17 patients at our institution. Four patients had minimally invasive mitral valve replacement. Mean patient age was 62.2 +/- 16.3 years; preoperative NYHA class was 3.06 +/- 0.2; ejection fraction was 64.1 +/- 14.7%. Echocardiography was performed pre-, intra- and postoperatively, and at 3-6, 12 and 24 months follow up. RESULTS: Fifteen patients had an uneventful intra- and postoperative course. Two patients died, one from acute left heart failure at 6 h after surgery, and one on the first postoperative day after resuscitation for ventricular fibrillation. A small-sized prosthesis was implanted in four patients, medium-sized in eight and large-sized in five. The mean duration of cardiopulmonary bypass was 138.3 +/- 37.0 min; mean cross-clamp time was 91.3 +/- 26.3 min. Postoperative control echocardiography showed a mean valve orifice area of 2.5 +/- 0.4 cm2, transvalvular velocity (Vmax) was 1.6 +/- 0.4 m/s, and mean pressure gradient 3.6 +/- 2.0 mmHg. Echocardiographic evaluation after 3, 6 and 12 months showed no significant difference compared with the intraoperative data. Three patients had a minor mitral regurgitation (grade I-II). At 12 months all patients were in NYHA class I or II. CONCLUSION: The implantation technique of the QMV is more demanding, but the prosthesis is a promising alternative to conventional biological mitral valve replacement. Further follow up is needed to confirm these favorable mid-term results. PMID- 11041183 TI - Immediate results of the 'Bioglis' valve use in the surgical treatment of Ebstein's anomaly. AB - BACKGROUND AND AIM OF THE STUDY: The optimum material for heart valves bioprostheses remains in dispute. We have created a biological valve made from hepatic Glisson's capsule and named 'Bioglis'. Experimental studies have shown the major characteristics of the 'Bioglis' valve to be equivalent to, and in some cases superior to, those of traditional valves made from xenopericardial tissue. We present the first experience of the 'Bioglis' valve use in surgical treatment of Ebstein's anomaly. METHODS: Twelve consecutive patients (age range: 7-48 years) with Ebstein's anomaly who underwent surgery between 1997 and 1999 were reviewed. A 'Bioglis' valve was implanted in all cases; two patients underwent repeat surgery because of incompetence of a previously implanted xenopericardial valve. The 'Bioglis' valve was formed, using a flexible frame, from the hepatic Glisson's capsule of bull calves. Short-term results at between two and 10 weeks after surgery were analyzed. Valvular function of the implanted 'Bioglis' valve was monitored by echocardiography. RESULTS: The implanted 'Bioglis' valve diameter ranged from 31 to 33 mm. There were no in-hospital deaths or complications. Echocardiography showed good function of the bioprostheses. Consequent peak and mean pressure gradients across the biological valve ranged from 3 to 7 mmHg and from 1.2 to 2.3 mmHg, respectively. Valve insufficiency occurred in one patient, but was minimal. CONCLUSION: Preliminary results with the 'Bioglis' support our recommendation of this new biological valve for clinical use. PMID- 11041184 TI - CarboMedics mechanical prosthesis: performance at eight years. AB - BACKGROUND AND AIM OF THE STUDY: The bileaflet St. Jude Medical mechanical prosthesis has been implanted for over 20 years. The purpose of this study was to evaluate the clinical performance of the bileaflet CarboMedics (CM) prosthesis, which was introduced in 1986. METHODS: The CM prosthesis was implanted in 1,258 patients (709 males, 549 females; mean age 60.9 +/- 12.3 years) between 1989 and 1997. The prosthesis distribution was aortic valve replacement (AVR) 613; mitral valve replacement (MVR) 447; and multiple replacement (MR) 231. Coronary artery bypass (CAB) was performed in 334 (26.6%) patients; previous procedures had been performed in 346 (27.5%). The age distribution was <60 years (n = 527), 61-70 years (n = 424) and >70 years (n = 307). Risk factors assessed were age or age groups, gender, CAB, previous surgery, rhythm, valve position, status and NYHA functional class. The total follow up was 4,765.0 patient-years (pt-yr), and was 98.4% complete. RESULTS: The early mortality rate was 5.6% (AVR 4.8%, MVR 3.7%, MR 11.5%). The late mortality rate was 3.7%/pt-yr (n = 174), and valve-related mortality 1.1%/pt-yr (n = 50). The total thromboembolism (TE) rate was 4.1%/pt-yr (n = 195) (p = NS by valve position); the major TE rate was 1.9%/pt-yr and fatal TE rate 0.31%/pt-yr (n = 15). The valve thrombosis rate was 0.31%/pt-yr (n = 15; 11 MVR, four MR). The fatal thrombosis rate was 0.06%/pt-yr (n = 3; two MVR, one MR). The hemorrhage rate was 2.7%/pt-yr (n = 128) and fatal hemorrhage rate 0.4%/pt-yr (n = 20). The reoperation rate was 1.0%/pt-yr (n = 46), fatal 0.1%/pt yr (n = 5). The actuarial freedom from overall TE at eight years was 77.3 +/- 2.8%; major TE 88.5 +/- 1.6%, and hemorrhage 76.4 +/- 3.2% (all p = NS by valve position). There were no independent predictors of overall TE and TE exclusion of early events. The only predictor for TE major was status (emergency > urgent > elective). The actuarial freedom from valve-related mortality at eight years was 91.4 +/- 1.8% (p = NS by position) (actual freedom 93.0 +/- 1.3%). The actuarial freedom from valve-related reoperation was 91.1 +/- 2.4% (p <0.05; AVR > MVR and MR, MVR > MR) (actual freedom 92.2 +/- 2.7%). Overall survival rate at eight years was 68.2 +/- 2.3% (p <0.05; AVR > MVR and MR, MVR > MR). CONCLUSION: The clinical performance of the CarboMedics mechanical prosthesis is satisfactory when implanted in the mitral, aortic and multiple positions. PMID- 11041185 TI - The Ultracor tilting disc heart valve prosthesis in the aortic position: mid-term follow up. AB - BACKGROUND AND AIM OF THE STUDY: The Ultracor valve is a recent introduction in the evolution of the tilting disc valve. This report summarizes a single surgeon's experience with the valve in the aortic position over a nine-year period. METHODS: Between 1990 and 1999, 94 patients received an Ultracor heart valve prosthesis in the aortic position. Forty-five patients (48%) had associated procedures, of which 32 (34%) were coronary artery grafts. Mean follow up was 2.6 years; total follow up was 229 patient-years. Follow up was 100% complete. RESULTS: The actuarial survival rate, including operative mortality rate, at five years was 82%. Actuarial freedom from valve-related death was 96% at five years. The linearized complication rate was 1.7%/year for late valve-related mortality, 1.7%/year for thromboembolism, 2.2%/year for anticoagulant-related hemorrhage (ACH), 1.3%/year for prosthetic valve endocarditis (PVE), and 1.7%/year for reoperation. The estimates of actuarial freedom from complications at five years were thromboembolism 89%, ACH 90%, PVE 96% and reoperation 96%. No structural failure or valve thrombosis was observed. CONCLUSION: Our experience over nine years showed the Ultracor heart valve prosthesis in the aortic position to be comparable with other currently evaluated mechanical heart valves in terms of durability and clinical results. Further study in this area should concentrate on the impact of valve type on left ventricular recovery post-implantation to provide additional information to the surgeon when selecting a valve from the plethora of choices available. PMID- 11041186 TI - The CarboMedics 'Top-Hat' aortic valve prosthesis: short-term results. AB - BACKGROUND AND AIM OF THE STUDY: The CarboMedics 'Top-Hat' aortic valve prosthesis has been specifically designed for supra-annular implantation. The aim of this study was to assess the safety of implantation of this prosthesis by reporting the short-term results of follow up. METHODS: Between May 1993 and May 1998, 128 patients (mean age 62.5 +/- 9.8 years; range: 22-76 years) received a CarboMedics 'Top-Hat' prosthesis at our institution. Among patients, 55% were in NYHA functional classes III or IV, and 54.7% had an isolated aortic valve replacement. Associated procedures were: coronary artery bypass grafting (25.7%), double valve replacement (17.1%), treatment of ascending aortic aneurysm (4.7%) and miscellaneous (5.5%). Follow up was 100% complete; total cumulative follow up was 265 patient-years (pt-yr) (range: 2-60 months). RESULTS: The overall mortality rate was 1.5% (two deaths). The operative mortality rate was 0.8% (one death); this patient died from neurological complications after operation for aortic dissection. The other patient died on postoperative day 40 from a massive cerebral hemorrhage. Four patients presented thromboembolic events; in all cases these were reversible ischemic neurologic deficits. One patient had a nonstructural deterioration (endocarditis) and required reoperation. Freedom from mortality was 98.3% at five years (linearized rate of 0.75%/pt-yr). Freedom from thromboembolism was 63.1% at five years (linearized rate 1.5%/pt-yr). CONCLUSION: Short-term results with the CarboMedics 'Top-Hat' prosthesis were satisfactory, with low rates of morbidity and mortality. As this prosthesis has demonstrated a good reliability to date, we have continued its implantation in our institution, and long-term follow up will be necessary to confirm these good early results. PMID- 11041187 TI - Aortic valve replacement for endocarditis: determinants of early and late outcome. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to determine risk factors for operative mortality, recurrent infection, reoperation and long-term survival following aortic valve replacement (AVR) for infective endocarditis. METHODS: Between 1973 and 1997, 109 patients (91 male, 18 female, mean age 52.6 years) underwent isolated AVR for infective endocarditis in our unit. Native valve endocarditis was present in 89 (81.6%) and prosthetic valve endocarditis in 20 (18.4%). Active culture-positive endocarditis was present in 53 (48.6%). Preoperatively, 99 patients (90.8%) were in NYHA classes III and IV. Indications for surgery included cardiac failure in 41 patients, valvular dysfunction in 38, vegetations in 18, sepsis in seven, abscess in six and embolism in four. Mechanical valves were implanted in 69 patients (63.3%) and bioprostheses in 40 (36.7%), including a homograft in 19 (17.4%). Follow up was complete (mean 5.8 years; range: 0-23.8 years; total 633.5 patient-years). RESULTS: The operative mortality was 10.1% (11 deaths). At ten years, freedom from recurrent infection was 94.2%, and freedom from reoperation 83.6%. Biological valve and younger age were significant adverse parameters for freedom from reoperation (p = 0.01 and p = 0.01). There have been 21 late deaths, 15 due to cardiac causes. Kaplan-Meier survival, including operative mortality, at five and ten years was 77.4% and 68.0%, respectively. On Cox proportional hazards regression, Staphylococcus aureus infection (p = 0.008) and older age (p = 0.04) were independent adverse predictors of survival. CONCLUSION: AVR for endocarditis carries a relatively high operative mortality, but can result in a satisfactory freedom from recurrent infection, reoperation and long-term survival. Analysis of our series demonstrates that implantation of a biological valve limits the freedom from reoperation and that infection by Staph. aureus reduces the probability of long term survival. PMID- 11041188 TI - Evaluation of serum cytokine concentrations in patients with infective endocarditis. AB - BACKGROUND AND AIM OF THE STUDY: Early diagnosis of infective endocarditis is important for clinical outcome, as mortality increases if diagnosis is delayed. Diagnosis is based on clinical features, echocardiography and blood culture findings, but negative blood cultures have been reported in 5-15% of proven cases. The study aim was to investigate serum cytokine levels in patients with infective endocarditis, and the possible use of these data in diagnosis and monitoring of the disease. METHODS: The study group comprised 40 patients with acquired rheumatic valvular heart disease and ongoing infective endocarditis. A diagnosis of infective endocarditis was established by clinical examination, echocardiography, laboratory investigations (inflammatory parameters) and positive blood cultures (n = 34). Two control groups included patients with acquired rheumatic valvular heart disease: 15 without infective endocarditis, and 15 with active urinary tract infection with significant bacteriuria. Serum interleukin-1alpha (IL-1alpha), interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) levels were measured on three occasions during antimicrobial treatment (mean period 14 +/- 7 days). RESULTS: Serum IL-1alpha and TNF-alpha levels were not elevated in the study group, or in controls (IL-1alpha <3.9 pg/ml; TNF-alpha <10 pg/ml). Serum IL-6 levels were elevated on all occasions in patients with infective endocarditis (first measurement: 37.0 +/- 44.3 pg/ml; second 18.7 +/- 16.4; third 8.5 +/- 5.2) with a significant tendency to decrease during treatment (p <0.01, ANOVA). In all controls without infection the serum IL 6 concentrations were below calibration range (<3.2 pg/ml). In the control group with active urinary tract infection, IL-6 concentrations were slightly (but not significantly) elevated (4.49 +/- 1.82 pg/ml, p = NS). CONCLUSION: Elevated serum IL-6 levels may suggest ongoing infective endocarditis and might be used to aid in diagnosis and monitoring of treatment of the disease. Serum IL-1alpha and TNF alpha levels were not affected. A further understanding of the role of serum cytokine concentrations in the diagnosis, prognosis and monitoring of infective endocarditis might be valuable in clinically uncertain diagnoses, especially when blood cultures are negative. PMID- 11041189 TI - Development and evaluation of a swine model to assess the preclinical safety of mechanical heart valves. AB - BACKGROUND AND AIM OF THE STUDY: The current standard of in vitro and in vivo preclinical heart valve testing has recently been questioned because of its failure to reveal the thrombogenic potential of the Medtronic Parallel prosthetic valve. The aim of this study was to develop a swine model for the in vivo preclinical evaluation of mechanical heart valves, and to assess the ability of this model to identify mechanical heart valve design features that result in valve-related thrombosis. METHODS: Twenty-two swine underwent mitral valve replacement (MVR) using three different bileaflet mechanical valve designs (St Jude Medical, CarboMedics, Medtronic Parallel). Each animal was placed in an anticoagulation protocol (group I, INR 3.0-3.5; group II, INR 2.0-2.5; group III, no anticoagulation) and followed for up to 20 weeks. Terminal studies were performed on all animals surviving for more than 30 days. RESULTS: Twenty-one animals survived the immediate postoperative period. Four of six group I animals died from hemorrhagic (large wound hematoma; hemopericardium) complications early in the study. In the two long-term (61 and 89 days) survivors, INRs of 3.0 to 3.5 were never achieved (61-day survivor, mean INR 2.0 +/- 1.03; range: 0.8-5.4; 89 day survivor, mean INR 1.92 +/- 1.34; range: 1.0-7.9). Pathological analysis of explants from group I survivors revealed minimally obstructive fibrous sheathing on the inflow orifice without restriction of bileaflet motion (61 and 89 days), and two large perivalvular defects (61 days). Six of seven group II animals died from early hemorrhagic complications (hemopericardium) (mean INR 2.32 +/- 1.84; range: 0.8-8.2). Vegetations resulting in obstruction of both sides of the valve orifice and restriction of bileaflet motion were observed in a group II survivor (mean INR 2.33 +/- 1.58; range: 0.9-7.0). Group III animals (n = 8) survived for a mean of 106 +/- 60 days (range: 1-177 days). In group III, fibrous sheathing was present on all explanted valves and organized thrombi in six valves; orifice obstruction (seven valves) and restriction of bileaflet motion (three valves) were also observed. CONCLUSION: The use of MVR in swine as a preclinical model to evaluate the safety and performance of mechanical heart valves is limited by: (i) difficulty in maintaining safe levels of anticoagulation with warfarin, resulting in a high incidence of hemorrhagic complications; (ii) marked fibrous sheath formation and associated thrombosis; and (iii) an increased incidence of perivalvular defects, believed to result from normal somatic growth occurring in young swine. PMID- 11041190 TI - Particle image velocimetry investigation of intravalvular flow fields of a bileaflet mechanical heart valve in a pulsatile flow. AB - BACKGROUND AND AIM OF THE STUDY: Our previous studies of bileaflet mechanical heart valves (MHV) explanted from sheep revealed patterns of localized platelet aggregation on valve surfaces, which may have clinical relevance. Since flow phenomena may promote localized platelet aggregation, an evaluation of flow within a valve lumen was conducted. METHODS: Phase-locked particle image velocimetry (PIV) measurements were obtained within the lumen of a 'mitral' model bileaflet MHV with transparent acrylic leaflets and housing, in a pulsatile flow loop. Instantaneous, two-dimensional flow maps of a central plane, perpendicular to the flow and leaflet pivot axes, were obtained at discrete times during the simulated cardiac cycle. Flow conditions were cardiac output, 3.5 l/min; rate, 72 beats/min; and systolic duration, 300 ms, using blood analog fluid refractive index-matched to acrylic. Leaflet closing velocities and angles were found using double-exposure imagery, and maximum leaflet closing velocity was extrapolated from regression analysis. RESULTS: During full opening, flow within the three lumenal orifices formed a three-peak axial velocity profile. Vorticity was concentrated in shear layers adjacent to downstream leaflet surfaces and in downstream wakes. Forward flow peak velocity was 90 cm/s, with a steep velocity gradient in the central orifice. During closing, the central-gap regurgitant flow formed a jet (peak velocity, 144 cm/s). High vorticity occurred near leaflet leading and trailing edges. During full closure, first a transient (<3 ms) 'stopping vortex' developed near the leaflet trailing edge, followed by a wall jet which formed at the leaflet-housing junction. Maximum leaflet closing velocity was 1.4 m/s. CONCLUSION: Localized jets, steep velocity gradients, high vorticity and vortex recirculation have been observed in vitro near model MHV surfaces. In vivo, each of these flow phenomena, when occurring near valve surfaces, may promote localized platelet aggregation. For the acrylic leaflets, maximum velocity was comparable with results reported for pyrolytic carbon leaflets. PIV of fully transparent models is a promising method for evaluating lumenal flows. PMID- 11041191 TI - Iatrogenic left ventricular-right atrial fistula following mitral valve replacement and tricuspid annuloplasty: diagnosis by transthoracic and transesophageal echocardiography. AB - Acquired left ventricle-to-right atrium communications are a known complication of valvular heart surgery. Previous reports have described the clinical features and diagnosis using cardiac catheterization. We report two cases of acquired left ventricle-to-right atrium fistula following mitral valve replacement. Particular emphasis is placed on the diagnosis using transthoracic and transesophageal echocardiography, obviating the need for cardiac catheterization before repair. PMID- 11041192 TI - Eighteen-year follow-up after Hancock II bioprosthesis insertion. PMID- 11041193 TI - The aortic valve blood supply. PMID- 11041194 TI - Chordal shortening: a technique to be set aside? PMID- 11041195 TI - Management of foot pain associated with accessory bones of the foot: two clinical case reports. AB - STUDY DESIGN: Case study. OBJECTIVES: To discuss the differential diagnosis, the nonsurgical and postoperative management of common accessory bones of the foot. BACKGROUND: Accessory bones of the foot that are formed during abnormal ossification are commonly found in asymptomatic feet. Two of the most common accessory bones are the accessory navicular and the os peroneum. Their painful presence must be considered in the differential diagnosis of any acute or chronic foot pain. The optimal treatment for the conservative and postoperative management of painful os peroneum and accessory navicular bones remains undefined. METHODS AND MEASURES: Therapeutic management of the fractured os peroneum included bracing, taping, and foot orthotics to allow healing of involved tissues, and stretching. The focus of the postoperative management of the accessory navicular was joint mobilization and progressive strengthening. Dependent variables included level of pain with provocation and alleviation tests of joint and soft tissue; girth and sensory tests of the foot and ankle; goniometric measures of foot and ankle; strength of ankle and hip muscles; functional tests; and patient's self-reported pain status. RESULTS: The patient with the fractured os peroneum was treated in 13 visits for 10 weeks. At discharge from physical therapy, the patient had the following outcomes relative to the noninvolved side: 100% return of normal sensation tested by light touch and vibration; pain decreased from 6/10 to 1/10; 100% reduction of swelling with ankle girth to normal; 100% range of motion of ankle and subtalar joints. Strength in plantar flexion and eversion remained 20% impaired (80% return to normal) secondary to pain. Upon discharge, he still reported mild pain when walking but was able to return to previous leisure activities. The second patient with the accessory navicular was treated in 18 visits over 9 weeks. Relative to the uninvolved side, she was discharged with the following: 70% return of range of motion in the foot and ankle, 100% of strength in hip and ankle, and 100% return of balance. She could squat and jump without pain and she returned to full premorbid activity level. CONCLUSIONS: Rehabilitative management of both cases addressed specific impairments and was successful in improving the patients' activity limitation. Clinicians should be aware that these accessory bones are possible sources of disability, secondary to foot pain. PMID- 11041196 TI - Effect of 10%, 30%, and 60% body weight traction on the straight leg raise test of symptomatic patients with low back pain. AB - STUDY DESIGN: Single group test-retest repeated measures. OBJECTIVES: To determine the effects of lumbar traction with 3 different amounts of force (10%, 30% and 60% body weight) on pain-free mobility of the lower extremity as measured by the straight leg raise (SLR) test. BACKGROUND: There are several recommendations on how lumbar traction should be performed, but the duration, frequency, force, and type of technique to be applied differ among the sources. METHODS AND MEASURES: Ten subjects with subjective complaints of low back pain or radicular symptoms with a positive unilateral SLR test below 45 degrees participated in this study. The pain-free mobility of the lower extremity in the SLR test position was measured prior to and immediately following 5 minutes of static traction in the supine position. Random assignment in the order of the amount of applied traction was implemented. RESULTS: The straight leg raise measurements were found to be significantly greater immediately following 30% and 60% of body weight traction as compared to pretraction and 10% of body weight traction. The mean (SD) SLR measurements were pretraction (24.1 degrees +/- 13.0), 10% of body weight traction (27.4 degrees +/- 14.5), 30% of body weight traction (34.0 degrees +/- 14.3), 60% of body weight traction (36.5 degrees +/- 15.8). CONCLUSIONS: The results of this study indicate that traction in this group of patients improved the mobility of the lower extremity during the SLR test. Both 30% and 60% of body weight tractions were shown to be effective for increasing motion beyond pretraction levels. PMID- 11041197 TI - Effectiveness of lateral slide exercise in an anterior cruciate ligament reconstruction rehabilitation home exercise program. AB - STUDY DESIGN: Two-group repeated measures design using a sample of convenience of subjects with anterior cruciate ligament (ACL) reconstructive surgery. OBJECTIVES: To determine the effect of incorporating one specific weight-bearing exercise (lateral slide exercise using a slide board) into an ACL reconstruction home exercise program. BACKGROUND: Reduced clinic visits have increased the importance of home exercise programs in knee ligament reconstruction rehabilitation. Few studies have been conducted to test the efficacy of specific exercises as part of a home-based treatment program on subjects who have undergone ACL reconstruction. METHODS AND MEASURES: Fourteen subjects who underwent patella tendon autograft reconstruction on one of their ACLs were studied. Testing consisted of the following 4 measurements: peak isometric knee extension torque, peak isometric knee flexion torque, maximum lateral step height, and lateral step-up repetitions to fatigue. Subjects were pretested at 8 weeks after surgery and were randomly placed into either a control or experimental group. The postsurgical rehabilitation was similar for both groups, except the experimental group incorporated lateral slide exercise into their home exercise program. All subjects were re-evaluated 14 weeks after surgery. RESULTS: A 2-way repeated measure ANOVA (group by test session), and posthoc testing revealed significant improvements in the slide group for quadricep strength (101.9 +/- 31.3 N m to 140.5 +/- 31.3 N m of torque), while the control group showed no significant increase (125.1 +/- 61.7 N m to 125.8 +/- 45.1 N m of torque). Lateral step height also improved in the slide group (from 22.9 +/- 5.3 cm to 28.7 +/- 5.6 cm), while the control group showed no increase (20.0 +/- 4.5 cm to 20.7 +/- 3.4 cm). Both groups increased in lateral step-up repetitions to fatigue. CONCLUSION: Including lateral slide exercise in a home exercise program after ACL reconstruction appears to improve knee extension strength. PMID- 11041198 TI - Comparison of first ray dorsal mobility among different forefoot alignments. AB - STUDY DESIGN: Experimental design using 1-way analysis of variance and regression analysis to test the influence of 3 forefoot alignments on the dorsal mobility of the first ray. OBJECTIVES: To determine the effect of forefoot alignment on the magnitude of first ray dorsal mobility to an imposed load and to describe any association between forefoot alignment and age on dorsal mobility of the first ray. BACKGROUND: Instability of the first ray has been implicated as a primary mechanical etiology of many foot problems. It has been proposed that a relationship exists between forefoot alignment and mobility of the first ray, with a varus aligned forefoot contributing to the development of an unstable first ray. METHODS AND MEASURES: Sixty female (n = 34) and male (n = 26) subjects aged 18-77 were assigned into valgus, neutral, and varus foot groups (20 per group) based on a clinical measurement of forefoot alignment. A load cell device measured dorsal mobility of the first ray under a standard load of 55 N. Within day repeat measures were taken from a subsample of subjects. In addition to reliability analysis, analysis of variance and regression analyses tested the relationship between forefoot alignment, age and sex, and mobility of the first ray. RESULTS: The forefoot valgus group demonstrated significantly less dorsal mobility of the first ray than neutral or varus groups. The varus and neutral groups were not significantly different from one another. Forefoot alignment and sex were significant linear predictors (R2 = 0.40) of first ray dorsal mobility. Age had no significant association to dorsal mobility of the first ray. CONCLUSION: Subjects having a valgus aligned forefoot had less dorsal excursion of the first ray than subjects having a neutral aligned forefoot. This investigation provides evidence supporting a relationship between forefoot alignment and mobility of the first ray. PMID- 11041199 TI - Tibialis posterior myofascial tightness as a source of heel pain: diagnosis and treatment. AB - STUDY DESIGN: We report 2 cases in which a novel tibialis posterior muscle stretch is used to treat heel pain and lower extremity impairment. OBJECTIVES: To explore dysfunction of the tibialis posterior as a source of heel pain. BACKGROUND: Heel pain is a common symptom of orthopaedic dysfunction of the lower extremity. Tibialis posterior tendon dysfunction is well documented in the medical and surgical literature, but its identification in its early or precursive stages has received little attention. METHODS AND MEASURES: An examination and treatment outline, incorporating a novel assessment and stretching technique, is presented. RESULTS: We identified a stage of dysfunction of the tibialis posterior ("Pre-Stage 1") without clinically identifiable tendon pathology. We refer to this as tibialis posterior myofascial tightness (TPMT). CONCLUSION: Tibialis posterior myofascial tightness is a clinical entity that may be differentially diagnosed in cases of heel pain and specifically treated. PMID- 11041200 TI - Okadaic acid suppresses TPA-induced differentiation by stimulating G1/S transition in human myeloblastic leukaemia ML-1 cells. AB - The association between the phosphorylation status of the retinoblastoma protein, pRb and changes in cell cycle control caused by either protein kinase C (PKC) or protein kinase A (PKA) stimulation was evaluated in human myeloblastic leukaemia ML-1 cells. TPA-induced PKC activation resulted in dephosphorylation of pRb and subsequently induced ML-1 differentiation based on morphological changes and CD14 expression. In the present study, we showed that inhibition of protein phosphatases (PP-1 and PP-2a) prevented the TPA-induced differentiation in ML-1 cells. Preinhibition of PP-1 and PP-2a activities with 1-100 nM okadaic acid dose dependently blunted the decrease in the phosphorylation status of pRb obtained with TPA and overrode cell cycle arrest. PKA stimulation with 8-chlorophenylthio cAMP (100 microM) decreased cell proliferation by 65% and the distribution of cells in the G1 phase significantly increased from 38% to 83% concomitant with a 34% decline in the number of cells present in the S phase. In addition, PKA stimulation significantly decreased the pRb phosphorylation status but did not elicit CD14 expression, indicating that cAMP-induced dephosphorylation of pRb cannot by itself trigger differentiation in ML-1 cells. PMID- 11041201 TI - Proliferation of glial cells in vivo induced in the neural lobe of the rat pituitary by lithium. AB - Lithium salts are widely used for treatment of psychiatric illness. Lithium also affects cell proliferation. During investigation of the effect of lithium chloride on the central nervous system (CNS) of nephrectomized rats, we noted numerous mitotic figures in the neural lobe of the pituitary. Morphologic criteria established that the mitotic cells were astrocytes, the supporting glial cells of the CNS, also known as pituicytes. Equimolar doses of chlorides of chemically related cations (sodium, potassium, rubidium) had no such effect. PMID- 11041202 TI - Tuning in the transcriptome: basins of attraction in the yeast cell cycle. AB - Image processing techniques and wavelet analyses have been applied to the yeast cell cycle expression microchip data to reveal large-scale temporally coherent structures and high frequency oscillations in mRNA levels through the cycle. Because transitions in expression frequently occur in phase, they appear as peaks or troughs in colour maps and contour plots of expression levels. Although apparent in the untreated data, these transitions were identified and enhanced by convolution of a Laplacian kernel with the expression arrays of the first 4096 genes. Transitions associated with maximum up- or down-regulation of mRNA levels appear as bands at 30-40 min intervals through two cell cycles. Time-frequency analyses using wavelet transforms support these visualization techniques and lead to the conclusion that, with respect to gene expression, the dominant period is not the cell cycle (90-120 min) but, more commonly, the higher frequency 30-40 minute submultiple of the cycle period. PMID- 11041203 TI - Forecasting the growth of multicell tumour spheroids: implications for the dynamic growth of solid tumours. AB - The growth dynamics of multicell tumour spheroids (MTS) were analysed by means of mathematical techniques derived from signal processing theory. Volume vs. time trajectories of individual spheroids were fitted with the Gompertz growth equation and the residuals (i.e. experimental volume determinations minus calculated values by fitting) were analysed by fast fourier transform and power spectrum. Residuals were not randomly distributed around calculated growth trajectories demonstrating that the Gompertz model partially approximates the growth kinetics of three-dimensional tumour cell aggregates. Power spectra decreased with increasing frequency following a 1/f(delta) power-law. Our findings suggest the existence of a source of 'internal' variability driving the time-evolution of MTS growth. Based on these observations, a new stochastic Gompertzian-like mathematical model was developed which allowed us to forecast the growth of MTS. In this model, white noise is additively superimposed to the trend described by the Gompertz growth equation and integrated to mimic the observed intrinsic variability of MTS growth. A correlation was found between the intensity of the added noise and the particular upper limit of volume size reached by each spheroid within two MTS populations obtained with two different cell lines. The dynamic forces generating the growth variability of three dimensional tumour cell aggregates also determine the fate of spheroid growth with a strong predictive significance. These findings suggest a new approach to measure tumour growth potential. PMID- 11041204 TI - Proliferation in murine epidermis after minor mechanical stimulation. Part 1. Sustained increase in keratinocyte production and migration. AB - It was our objective to obtain an insight into the details and dynamics of the cell proliferative changes following minor barrier disruption, the mechanisms of recovery, and their regulation. Hair of the dorsal area of DBA2-mice was removed and the epidermis was tape stripped. Tritiated thymidine was injected into groups of mice at daily intervals thereafter. Labelling and nuclear densities were measured at several time intervals later in the various epidermal strata to characterize cell production and cell fluxes through the tissue. A dramatic proliferative response was observed at 24 h when the labelling density increased more than sixfold in the basal layer. Labelled cells rapidly appeared in suprabasal layers within a few hours in large quantities while this process took over 2 days in normal skin. Some cycling cells were also found in the suprabasal layer (pulse labelling at 24 h) in contrast with the controls. The cellular flux through the suprabasal layers was drastically (20-fold) increased and the transit time was shortened. Although the nuclear density in the basal layer showed only moderate changes it increased four-fold in the suprabasal layer within 5 days. A kinetic model analysis suggested that the cell cycle time of proliferative cells dropped from a normal value of about 200 h to less than 12 h post tape strip. After 7 days, the proliferative activation still persisted, even though at 3 days post tape strip the stratum corneum had been re-established. Hence, a mild mechanical alteration with removal of some parts of the cornified layer in mouse backskin epidermis triggers a huge proliferative response with massive overproduction of cells that lasts at least 7 days. Our findings suggest that the re-establishment of the cornified layer does not immediately shut down cell proliferation and that more complex, slower (long-term) regulatory processes are involved. PMID- 11041205 TI - Proliferation in murine epidermis after minor mechanical stimulation. Part 2. Alterations in keratinocyte cell cycle fluxes. AB - We have recently shown that a mild mechanical irritation (tape strip) of the epidermis on the back skin of adult mice induces a strong and long lasting increase in proliferative activity and cell production. This was revealed by following the fate of 3HTdR-pulse labelled cells within the basal and suprabasal layers. To obtain further insight into the dynamics of cell kinetic changes we also performed double labelling experiments with 3HTdR and BrdUrd at various times after tape stripping. The technique for analysing the data had to account for a non stationary cell flux. A novel biometrical technique was developed which provides parameter estimates on the S-phase duration, the cell cycle duration and a parameter characterizing the degree of nonstationarity. When applied to the mechanically irritated epidermis we observed that the cell flux through the S phase in the basal layer was accelerated by a factor of 10 between 18 and 36 h post tape strip. This activation declined slightly in the subsequent days and remained 4-6 fold higher than in the normal steady state for over 7 days post tape strip. The duration of the S-phase was 3-5 h and showed little variation. We conclude that mild mechanical irritation only affecting the stratum corneum has major stimulatory effects on the cell kinetics of proliferative keratinocytes in the basal layer of the epidermis indicating the existence of a powerful regulatory mechanism. PMID- 11041206 TI - The regulation of motile activity in fish chromatophores. AB - Chromatophores, including melanophores, xanthophores, erythrophores, leucophores and iridophores, are responsible for the revelation of integumentary coloration in fish. Recently, blue chromatophores, also called cyanophores, were added to the list of chromatophores. Many of them are also known to possess cellular motility, by which fish are able to change their integumentary hues and patterns, thus enabling them to execute remarkable or subtle chromatic adaptation to environmental hues and patterns, and to cope with various ethological encounters. Such physiological color changes are indeed crucial for them to survive, either by protecting themselves from predators or by increasing their chances of feeding. Sometimes, they are also useful in courtship and mutual communications among individuals of the same species, leading to an increased rate of species survival. Such strategies are realized by complex mechanisms existing in the endocrine and/or nervous systems. Current studies further indicate that some paracrine factors such as endothelins (ETs) are involved in these processes. In this review, the elaborate mechanisms regulating chromatophores in these lovely aquatic animals are described. PMID- 11041207 TI - The tyrosinase gene and oculocutaneous albinism type 1 (OCA1): A model for understanding the molecular biology of melanin formation. AB - Through the last century there has been a steady progression in our understanding of the biology of melanin biosynthesis. Much of this work includes the analysis of coat color mutations of the mouse and albinism in man. Our understanding has been greatly enhanced in the last 10 years, as the molecular pathogenesis of albinism has been better understood. Different mutations of the tyrosinase gene (TYR) , and their association with oculocutaneous albinism type 1 (OCA1) has provided insight into the biology of tyrosinase, including protein trafficking and structure/function analysis. Several questions still remain, including cryptic mutations that affect tyrosinase activity and the minimum amount of pigment required for normal optic development. The next 10 years should prove just as exciting as the last. PMID- 11041208 TI - Controlling gene expression in mice with tetracycline: application in pigment cell research. AB - Genetic manipulation techniques are widely used in mice to study the functions of genes. The most common strategy for assessing in vivo function involves making irreversible changes in the genome by homologous recombination. To complement this approach, a number of systems have been developed that allow specific and controlled expression of a gene. One of the more versatile and promising systems is based on the tetracycline (tet) responsive bacterial tetracycline repressor (TetR). In recent years, the tet system has proven to be a valuable method for understanding the function of genes involved in a number of physiological processes, including mouse models for human diseases such as cancer and neurological and pigment disorders. This review will highlight the power and elegance of the tet system by focusing on its utility in the study of two pigment cell-related biological problems, the pathogenesis of melanomas and melanocyte development in the embryo. PMID- 11041209 TI - Small Gtpase rab3A is associated with melanosomes in melanoma cells. AB - Rab3A is a small guanosine triphosphate (GTP)-binding protein that has been recently implicated in intracellular vesicle transport and the secretion of neurotransmitters in neuronal cells. We demonstrate here that Rab3A is associated with melanosomes in pigment cells. Rab3A as well as Rabphilin3A, a putative target protein of Rab3A, were detected in the melanosome fraction, purified from B16 murine melanoma cells by sucrose density gradient ultracentrifugation. In contrast, Rab GDP dissociation inhibitor (GDI), a GDP/GTP exchange protein for Rab3A, was found in the cytosol fraction. Further studies using confocal laser scanning microscopy and immunoelectron microscopy revealed that immunoreactive Rab3A is localized in conjunction with the melanosomal membrane. These results suggest the possibility of involvement of Rab3A-Rabphilin3A complex, regulated by Rab GDI, in the intracellular transport of melanosomes in pigment cells. PMID- 11041210 TI - Mutant alleles at the brown locus encoding tyrosinase-related protein-1 (TRP-1) affect proliferation of mouse melanocytes in culture. AB - Tyrosinase related protein-1 (TRP-1) is a melanocyte-specific gene product involved in eumelanin synthesis. Mutation in the Tyrp1 gene is associated with brown pelage in mouse and oculocutaneous albinism Type 3 in humans (OCA3). It has been demonstrated that TRP-1 expresses DHICA oxidase activity in the murine system. However, its actual function in the human system is still unclear. The study was designed to determine the effects of mutation at two Typr1 alleles, namely the Tyrp1b (brown) and Tyrp1b-cj (cordovan) compared with wild type Tyrp1B (black) on melanocyte function and melanin biosynthesis. The most significant finding was that both of the Tyrp1 mutations (i.e. brown expressing a point mutation and cordovan expressing decreased amount of TRP-1 protein) resulted in attenuation of cell proliferation rates. Neither necrosis nor apoptosis was responsible for the observed decrease in cell proliferation rates of the brown and cordovan melanocytes. Ultrastructural evaluation by electron microscopic analysis revealed that both mutations in Tyrp1 affected melanosome maturation without affecting its structure. These observations demonstrate that mutation in Tyrp1 compromised tyrosinase activity within the organelle. DOPA histochemistry revealed differences in melanosomal stages between black and brown melanocytes but not between black and cordovan melanocytes. There were no significant differences in tyrosine hydroxylase activities of tyrosinase and TRP-1 in wild type black, brown and cordovan melanocyte cell lysates. We conclude that mutations in Tyrp1 compromise cell proliferation and melanosomal maturation in mouse melanocyte cultures. PMID- 11041211 TI - A novel approach to gene therapy of albino hair in histoculture with a retroviral streptomyces tyrosinase gene. AB - In order to induce melanin production in mammalian cells with pigment disorders such as albino hair, a recombinant retrovirus containing the mel locus of Streptomyces antibioticus was constructed. The S. antibioticus mel locus, which consists of the open reading frame (ORF)-438 and the tyrosinase gene, was specifically derived by polymerase chain reaction (PCR) from Streptomyces plasmid pIJ702. The ORF-438 is required for the transfer of copper to apotyrosinase, which is essential for tyrosinase enzymatic activity. The tyrosinase gene was inserted into the XhoI/BamHI cloning site of the pLXSN retroviral vector to obtain pLtyrSN. An internal ribosome entry site (IRES) suitable for mammalian cell expression was obtained from the pLXIN retroviral vector by PCR. The ORF-438 and IRES DNA fragments were inserted into the pLtyrSN vector to obtain the tyrosinase-expression retroviral vector pLmelSN. The expression vector was amplified in murine PT67 packaging cells, where the ORF-438 and tyrosinase genes were also co-expressed as determined by reverse transcription-PCR. In order to evaluate the vector's ability to restore pigment production in cells with a pigment disorder, albino-mouse skins were histocultured and then infected with the pLmelSN retrovirus. Six days after infection, melanin granules were observed in approximately 60% of albino-mouse hair follicles in the histocultured skin. These results demonstrated that the S. antibioticus mel operon could express an active tyrosinase and produce melanin in the albino-mouse hair follicles. This novel gene therapy approach, using a small and simple tyrosinase operon in a high expression vector, has a potentially wide application for therapy of pigment disorders in hair follicles. PMID- 11041212 TI - Up-Regulation and redistribution of Bax in ultraviolet B-irradiated melanocytes. AB - Ultraviolet B (UVB) radiation may activate or deteriorate cultured human epidermal melanocytes, depending on the doses and culture conditions. It is also reported that cultured human epidermal melanocytes derived from different pigmentary phenotypes showed different responses to UVB radiation. In this study, we examined whether apoptosis of melanocytes can be induced by physiologic doses of UVB irradiation using cultured human epidermal melanocytes derived from oriental males of skin types III and IV. Propidium iodide staining for DNA condensation and flow cytometric analyses demonstrated the apoptotic cell death of melanocytes following UVB irradiation (0-30 mJ/cm2). The levels of p53, Bax, and Bcl-2, determined by immunoblotting, revealed a dose-dependent increase in p53 and Bax, but the level of Bcl-2 remained unchanged. Confocal microscopic examination showed that Bax moved from a diffuse to a punctate distribution after UVB irradiation. However, there were no changes in the pattern of distribution of Bcl-2. These data suggest that the high constitutional level of Bcl-2 may protect melanocytes from UVB-induced injury, and that apoptotic death of melanocytes may be induced by the elevation and redistribution of Bax. PMID- 11041213 TI - Involvement of nitric oxide in UVB-induced pigmentation in guinea pig skin. AB - Ultraviolet (UV) B irradiation evokes erythema and delayed pigmentation in skin, where a variety of toxic and modulating events are known to be involved. Nitric oxide (NO) is generated from L-arginine by NO synthases (NOS). Production of NO is enhanced in response to UVB-stimulation and has an important role in the development of erythema. NO has recently been demonstrated as a melanogen which stimulates melanocytes in vitro, however, no known in vivo data has been reported to support this finding. In this study, we investigated the contribution of NO with UV-induced pigmentation in an animal model using an NOS inhibitor. UVB induced erythema in guinea pig skin was reduced when an NOS inhibitor, L-NAME (N nitro-L-arginine methylester hydrochloride), was topically applied to the skin daily, beginning 3 days before UVB-irradiation. Delayed pigmentation and an increased number of DOPA-positive melanocytes in the skin were markedly suppressed by sequential daily treatment with L-NAME. Furthermore, melanin content 13 days after UVB-irradiation was significantly lower in skin treated with L-NAME than in the controls. In contrast, D-NAME (N-nitro-D-arginine methylester hydrochloride), an ineffective isomer of L-NAME, demonstrated no effect on these UV-induced skin responses. These results suggest that NO production may contribute to the regulation of UVB-induced pigmentation. PMID- 11041214 TI - Mutational analysis of the modulation of tyrosinase by tyrosinase-related proteins 1 and 2 in vitro. AB - The albino (tyrosinase, Tyrc), brown (tyrosinase-related protein 1, Tyrp1b) and slaty (tyrosinase-related protein 2, tyrp2slt) loci are all involved in the regulation of melanogenesis. Phenotypes of inbred mice mutant at two or more of these loci are not always explicable by simple summation of the established or suspected catalytic functions of the gene products. These phenotypes suggest that relationships among the proteins extend beyond the obvious fact that they catalyze different steps in the same melanogenic pathway, and that they may also interact intimately in such a way that a mutation in one impacts the function of the other(s). Previous studies have attributed catalytic activities to each member of this trio; however, it has been difficult to study the proteins individually, either in vivo or in tissues or cells. Therefore, we undertook to transfect the genes, in revealing combinations, into COS-7 cells (which have no melanogenic apparatus of their own) to clarify the interacting functions of their encoded proteins. Specifically, we attempted to evaluate the effects of Tyrp1 and Tyrp2 proteins on tyrosinase protein. We report evidence that Tyrp1 stabilizes tyrosinase, confirming previous observations, and, in addition, demonstrate that Tyrp1 decreases tyrosinase activity. By contrast, Tyrp2 increases tyrosinase activity by stabilizing the protein. We conclude that both Tyrp1 and Tyrp2, in addition to other catalytic functions they may possess, act together to modulate tyrosinase activity. PMID- 11041215 TI - A functional model for progestogen-induced breakthrough bleeding. AB - During the past 5 years, a number of important advances have been made in our understanding of the mechanisms of sex steroid-induced breakthrough bleeding (BTB). These observations suggest that superficial endometrial vascular fragility may be the mechanism underlying BTB, and molecular changes in the microvasculature as well as hysteroscopic observations have supported this hypothesis. This paper aims to present a unified picture of our current understanding of BTB, particularly that associated with progestogens, to indicate current gaps in our knowledge and possible directions for future research. PMID- 11041216 TI - Disturbances of endometrial bleeding with hormone replacement therapy. AB - Breakthrough bleeding is a common problem in postmenopausal women taking hormone replacement therapy (HRT) and is often the single most important factor deterring women from continuing to use HRT, or from starting it in the first place. The mechanisms which underlie this unscheduled bleeding are poorly understood. The benefits of HRT in terms of longevity and quality of life are becoming increasingly apparent, and a greater understanding of why this bleeding occurs and how we can prevent or treat it, will undoubtedly enable more women to reap the potential considerable benefits of long-term oestrogen and progestogen replacement. What sets postmenopausal women apart from their counterparts in the mid-reproductive years is the increased likelihood of endometrial adenocarcinoma in which unscheduled bleeding is the presenting symptom. Therefore, spontaneous postmenopausal bleeding must always be appropriately evaluated. Hence, the occurrence of unscheduled bleeding with HRT may provide a dilemma with diagnosis as well as a challenge to acceptability. Combined HRT regimens tend to be predominantly progestogenic, and there is increasing evidence to suggest that some of the vascular changes seen in women taking long-term, low-dose progestogen only contraceptives may also occur in women taking HRT. PMID- 11041218 TI - Management of vaginal bleeding irregularities induced by progestin-only contraceptives. AB - As the use of progestin-only methods of contraception continues to increase worldwide, the problem of the vaginal bleeding disturbances these methods induce is becoming of increasing public health relevance. A number of approaches are used by clinicians to control these bleeding irregularities but few treatments have been adequately tested and, to date, none appears sufficiently effective. Better understanding of the mechanisms underlying vaginal bleeding as well as of the attitudes of women towards menstrual disturbances is needed, so that effective and acceptable therapies can be devised. PMID- 11041217 TI - Oxidative stress, vitamin E and progestin breakthrough bleeding. AB - Endometrial bleeding problems can be the major reason for discontinuing progestin only contraception. In this study the endometrial angiogenic response in Norplant users was found to be lower than in women with normal menstrual cycles. These disturbances in angiogenic response may be caused by oxidant-antioxidant imbalance in the endometrium. The aims of this study were to investigate the effect of progestin only contraceptives on blood concentrations of lipid peroxide and vitamin E, and the effect of vitamin E supplementation on endometrial angiogenic response in vitro. The subjects for this study were Norplant users, depo-medroxyprogesterone acetate (DMPA) users, and controls. Circulating lipid peroxide and vitamin E concentration was measured by routine methodology. Endometrial angiogenic response was assayed using an endothelial cell migration assay. The results showed that the blood concentrations of lipid peroxide from Norplant users with bleeding problems were significantly higher than normal menstrual controls (P < 0.05) and supplementation of vitamin E (in vitro) increased the endometrial angiogenic score. Blood concentrations of lipid peroxide were significantly increased (P < 0.05), and the blood concentrations of vitamin E were significantly decreased (P < 0.05) after 3 months exposure to Norplant or DMPA. The endometrial angiogenic scores in Norplant and DMPA users were significantly lower than in controls (P < 0.02). It is concluded that in progestin-only contraceptive users, higher lipid peroxide and lower vitamin E concentration may cause endometrial cell damage and decrease the endometrial angiogenic response. It is suggested that vitamin E supplementation may counteract these unwanted side-effects. PMID- 11041219 TI - The role of selective oestrogen receptor modulators in the treatment of endometrial bleeding in women using long-acting progestin contraception. AB - This paper explores the concept that endometrial breakthrough bleeding results from the stimulatory effects of oestrogen in the endometrium. Though 'progestin only' contraceptive regimens have long been associated with user dissatisfaction because of unpredictable vaginal bleeding, it is likely that the substantial contribution of endogenous ovarian oestradiol during such treatments predisposes the bleeding problems. Oestrogen causes endometrial proliferation, hyperplasia and neoplasia if unopposed. Oestrogen allows production of growth factors supporting angiogenesis which results in an abundance of dilated or fragile endothelial surface blood vessels, predisposing this tissue to bleeding when these vessels lose competence. PMID- 11041220 TI - Endothelial cell dysfunction following prolonged activation of progesterone receptor. AB - Progestin-only contraceptives are associated with breakthrough bleeding in up to 50% of users. The causes of blood vessel rupture are not well understood. Here we report that both normal and Norplant-exposed endothelium express progesterone receptor. Experiments performed in vitro on endothelial cells isolated from human endometrium revealed that longterm progesterone exposure leads to suppression of endothelial cell proliferation, inhibition of migration and alteration in the profile of extracellular matrix proteins secreted by human endometrial endothelial cells. In addition, we detected increased levels of matrix metalloproteinase-9 in endothelial cultures treated with progesterone. The effect of progesterone on the cell cycle, along with the increased amounts of matrix degrading enzymes, could account for breakdown of basement membrane components, vascular fragility and consequent vessel rupture leading to breakthrough endometrial bleeding. PMID- 11041221 TI - Heterogeneity of progesterone receptors A and B expression in human endometrial glands and stroma. AB - The human progesterone receptor (PR) is expressed as two isoforms, PRA and PRB, which function as ligand-activated transcription factors. In-vitro studies suggest that the isoforms differ functionally and that their relative expression in a target cell may determine the nature and magnitude of response to progesterone. We have shown recently that PRA and PRB are co-expressed in target cells of the human endometrium. The purpose of this study was to investigate the homogeneity of expression of PRA and PRB in target cells of the human uterus throughout the menstrual cycle. In the functionalis, PRA and PRB were expressed in comparable levels in glandular epithelium during the proliferative phase of the cycle, whereas there was persistence of PRB but not PRA in the glands during mid-secretory phase. In the stroma, there was predominance of the PRA isoform throughout the cycle. There was remarkable homogeneity in the relative expression of PRA and PRB in adjacent cells within the same tissue compartment, suggesting that the mechanisms regulating relative PR isoform expression are similarly active in these cells. By contrast, heterogeneity between glands was observed under some circumstances in the functionalis of the endometrium, suggesting PR isoform down-regulation by progesterone to be asynchronous. Heterogeneity was also seen between the glands of the basalis and functionalis of the endometrium implying region-specific responses to hormonal stimuli. This study demonstrates adjacent cell homogeneity in the relative expression of PRA and PRB in normal human endometrial tissue and a differential response to ovarian steroid hormones between cell types and between different regions within the same tissue. PMID- 11041222 TI - The structure of endometrial microvessels. AB - Recent studies have provided substantial evidence to highlight abnormalities in the structure of endometrial microvessels in users of progestogen-only contraception. Structural changes alone are unlikely to lead to breakthrough bleeding, but appear to be associated with a reduction in vessel integrity, and may reflect alterations in the control and growth of endometrial microvessels in those exposed to exogenous progestogens. In users of low-dose progestogens, immunohistochemical studies have demonstrated changes in superficial endometrial vascular morphology, density and in endometrial migratory cell populations. In Norplant users the endometrial endothelial basal lamina is deficient in the initial months of exposure, when bleeding problems are most common. The basal lamina refers to the very thin structure consisting mostly of collagen IV and laminin seen underlying endothelial and epithelial cells at the electron microscope level. In addition, hysteroscopic studies have demonstrated increased fragility of superficial vessels. Changes in endometrial microvascular anatomy associated with normal menstruation have been reviewed, and are compared with those seen following contraceptive steroid exposure. Likely mechanisms of breakthrough bleeding such as increased superficial vascular fragility and the possible alterations in endometrial vascular structure leading to this are discussed. PMID- 11041223 TI - Ovarian steroid and cytokine modulation of human endometrial angiogenesis. AB - A key mechanism underlying the cyclical growth of the endometrium is its ability to regenerate a vascular capillary network. In normal cycling human endometrium, angiogenesis is influenced by both endocrine and paracrine factors. Hormonal manipulation of the endometrium, such as that occurring during the use of steroidal contraception, appears to result in capillary proliferation and fragility. As a consequence of these vascular changes, contraceptive users may be predisposed to unpredictable uterine bleeding, which is responsible for the high frequency of contraceptive discontinuation. In this paper we address mechanisms responsible for vascular endothelial cell proliferation in normal and contraceptive steroid-exposed endometria. We propose that regulation of endometrial angiogenesis is mediated indirectly, via steroid and cytokine actions on vascular endothelial growth factor (VEGF), and we present data indicating that VEGF expression in normal endometrial stromal cells is increased by oestrogens and progestins. Three proinflammatory cytokines with angiogenic effects in other systems (i.e. interleukin-1beta, tumour necrosis factor-alpha and interferon gamma) do not appear to up-regulate VEGF expression in normal endometrial stromal cells. Well-characterized in-vitro models in conjunction with immunohistochemistry provide useful experimental systems to study endometrial neovascularization under physiological conditions and in those potentially perturbed via the use of contraceptive steroids. PMID- 11041224 TI - Perivascular smooth muscle alpha-actin is reduced in the endometrium of women with progestin-only contraceptive breakthrough bleeding. AB - It has been shown that the endometrium of women using progestin-only contraceptives has increased vascular fragility, although the structural basis for this weakness is unknown, as is its role in breakthrough bleeding (BTB). Perivascular cells such as pericytes and vascular smooth muscle cells surround capillaries during the maturation process following angiogenesis, and act to strengthen and stabilize the vessels. The aim of the present study was to quantify endometrial perivascular smooth muscle alpha-actin (alphaSMA) expression in women using Norplant with and without BTB problems, and compare it to controls. Using immunohistochemical techniques, vessels were classified as level 0, 1 or 2 depending on whether perivascular alphaSMA was absent, discontinuous or continuous. In 15 controls the subepithelial plexus had significantly more level 0 vessels than either the functionalis or basalis (61 +/- 4 versus 31 +/- 6 and 37 +/- 4%, P = 0.0006 and P = 0.0007 respectively). In contrast the functionalis and basalis had significantly more level 2 vessels than the subepithelial plexus (20 +/- 3 and 23 +/- 2 compared to 4 +/- 1%, P = 0.0005 and P = 0.000 respectively). The major finding of the study was that in Norplant users, where the relatively atrophic endometrium cannot be divided into different regions, women with BTB problems (n = 20) had significantly more level 0 vessels than those with reduced bleeding (n = 17) (60 +/- 4 versus 46 +/- 4%, P = 0.0302). Norplant users with BTB problems also had a non-significant reduction in level 2 vessels compared to women without bleeding problems (4 +/- 2 versus 11 +/- 4%, P = 0.0667). These results demonstrate that perivascular alphaSMA is reduced around the endometrial vessels of Norplant users with BTB compared to those with no bleeding problems, and strongly support the concept that reduced vascular structural integrity plays a key role in endometrial BTB. PMID- 11041225 TI - The effect of progestins on vascular endothelial growth factor, oestrogen receptor and progesterone receptor immunoreactivity and endothelial cell density in human endometrium. AB - One common side-effect of contraceptive use is that it often leads to disrupted endometrial bleeding patterns. This may be due to changes in endothelial density and vessel integrity. To investigate whether the level of endometrial immunoreactive vascular endothelial growth factor (VEGF), oestrogen receptor or progesterone receptor (PR) have any role in this, women were treated with either Mircette, a monophasic oral contraceptive, or Implanon, a long-acting gestagen, and immunohistochemistry performed. In addition a small number of endometria were studied from women treated with levonorgestrel released from an intrauterine coil. During the untreated normal cycle, there was a significant increase in glandular VEGF immunoreactivity and a significant decrease in PR immunoreactivity in the midand late secretory phases compared to the proliferative phase. There was a significant positive correlation between stromal VEGF immunoreactivity and endothelial cell density. This correlation was also apparent during treatment with Implanon, but not with Mircette. Disrupted bleeding patterns were associated with Implanon and to a lesser extent with Mircette. Both contraceptives significantly reduced glandular VEGF immunoreactivity but the intrauterine treatment with levonorgestrel resulted in strong glandular epithelial staining and intense staining of decidualized stromal cells. Implanon significantly increased glandular PR staining, but Mircette significantly reduced stromal PR staining when compared to secretory phase before-treatment biopsies. There were no changes in endothelial cell density or glandular or stromal ER during the normal cycle, or with use of either contraceptive. There was no association of the parameters measured with bleeding patterns or histological category. PMID- 11041226 TI - Role of nitric oxide in implantation and menstruation. AB - Nitric oxide (NO) is a major paracrine mediator of various biological processes, including vascular functions and inflammation. In blood vessels, NO is produced by the low-input constitutive endothelial NO synthase (eNOS) and is a potent vasodilator and platelet aggregation inhibitor. The inducible NOS isoform (iNOS) is capable of producing NO at high concentrations which have pro-inflammatory properties. Immunohistochemical and molecular studies of endometrial NOS expression, as well as animal experiments with NOS inhibitors, indicate that NO plays an important role in endometrial functions such as endometrial receptivity, implantation and menstruation. In rodents, both iNOS and eNOS are highly up regulated in the implantation sites, and NOS inhibitors show synergistic effects with antiprogestins in inhibiting the establishment of pregnancy. In the human endometrium, eNOS have been localized in the glandular epithelium and in endometrial microvascular endothelium, primarily during the luteal phase. iNOS has been found in the endometrial epithelium during menstruation, in immunocompetent endometrial cells, and in decidualized stromal cells. In primates, NO may be involved in the initiation and maintenance of menstrual bleeding by inducing tissue breakdown and vascular relaxation as well as by inhibiting platelet aggregation. Endometrium-derived NO may also play a role in myometrial relaxation during menstruation. These studies open up new applications for NO-donating and -inhibiting agents in uterine disorders. NO donors may be useful in the treatment of dysmenorrhoea and for promoting fertility. Antiprogestins, progesterone receptor modulators and iNOS inhibitors may find applications in the treatment and prevention of abnormal uterine bleeding. PMID- 11041227 TI - Regulation of matrix metalloproteinases in human endometrium. AB - Considerable evidence supports a role for matrix metalloproteinases (MMP) in menstruation, but their focal pattern of expression within perimenstrual and menstrual endometrium suggests local rather than hormonal regulation. Menstruation shares a number of features with inflammatory responses, with leukocyte infiltration, proliferation and activation, occurring in the endometrium prior to menstruation. We propose that the leukocytes release MMP at this time and also that interactions between leukocytes and the stromal and epithelial cells of the endometrium induce and activate MMP. Co-culture studies using mast cells or neutrophils with endometrial stromal cells support this hypothesis. How leukocytes enter the endometrium is not understood but a role for chemokines has been proposed. The expression patterns of eotaxin and its receptor CCR3 in endometrium support a role in chemoattraction of eosinophils but expression of monocyte chemotactic proteins 1 and 2 does not correlate with macrophage numbers. Nothing is known of how the leukocytes become activated. Nevertheless, the overall result is a tissue in which an inflammatory-type reaction occurs with release of a myriad of potent regulators. These induce production and activation of MMP and alter the ratio between these and their tissue inhibitors, resulting in tissue breakdown. PMID- 11041228 TI - Circulating sex hormones and endometrial stromelysin-1 (matrix metalloproteinase 3) at the start of bleeding episodes in levonorgestrel-implant users. AB - Unpredictable endometrial bleeding is the major side-effect of levonorgestrel releasing s.c. implants (Norplant), otherwise a method of choice for long-term contraception. The mechanisms responsible for bleeding are still unknown and no reliable treatment is available. Several matrix metalloproteinases (MMP) are expressed and activated in human endometrium only at menstruation and specific synthetic inhibitors of MMP fully prevent the tissue breakdown that occurs in menstrual-like endometrial explants. To investigate whether MMP are inappropriately expressed and activated in Norplant-treated endometria during bleeding episodes, volunteers were recruited to provide blood and endometrial biopsies at the start of bleeding episodes and during non-bleeding intervals. Whereas serum concentrations of levonorgestrel and sex hormones showed no change at bleeding, except for a slight decrease of oestradiol concentration, the expression and activation of stromelysin-1 released by explants cultured for 1 day were consistently increased at the start of bleeding episodes. Furthermore, stromelysin-1 was immunolocalized in stromal cells within breakdown areas of several bleeding endometria, but not in non-bleeding endometria. These observations suggest that the expression and activation of stromelysin-1 participate in the initiation of bleeding episodes upon Norplant contraception. New strategies in the prevention and treatment of abnormal bleeding based on MMP control should be envisaged. PMID- 11041229 TI - The role of matrix metalloproteinases and leukocytes in abnormal uterine bleeding associated with progestin-only contraceptives. AB - Progestin-only contraceptives are associated with menstrual bleeding disturbances; a major reason why these agents are discontinued. The pathogenesis of abnormal uterine bleeding associated with progestin-only contraceptives remains ill-defined. Matrix metalloproteinases (MMPs) and leukocytes are postulated to be involved in the process of normal menstruation. Immunolocalization of MMPs and leukocytes in (Norplant), and injectable depot medroxyprogesendometrium from women using the progestinterone acetate (DMPA), are widely used, safe and only contraceptives, Norplant or depot medroxyprogesterone acetate (DMPA) compared with normal controls, revealed foci of positive MMP-1 and -3 immunostaining in stromal cells and adjacent extracellular matrix, the presence of MMP-9 in various subtypes of leukocytes and alterations in mast cell phenotype. In women using progestin-only contraceptives, extent of endometrial MMP, neutrophil and eosinophil immunolocalization and the mast cell activation state was similar to or greater than that observed in perimenstrual control women. However, differences in MMP immunostaining were observed in endometrial samples from women using different progestin-only contraceptive agents; in particular, significantly higher MMP-1 immunostaining was observed associated with the use of Norplant compared with DMPA. No correlation was observed with the number of bleeding days recorded. These results suggest that MMP and leukocytes may be involved in endometrial breakdown in women using progestin-only contraceptives. PMID- 11041230 TI - The role of tissue factor in regulating endometrial haemostasis: implications for progestin-only contraception. AB - Abnormal uterine bleeding accounts for the unacceptably high discontinuation rate of progestin-only contraceptives. Previously, we found that in-vivo and in-vitro decidualization of human endometrial stromal cells was associated with elevated concentrations of tissue factor (TF), the primary initiator of haemostasis. Moreover, enhanced TF expression required progesterone receptor (PR) and epidermal growth factor receptor (EGFR) mediation. In the current study, endometrial biopsies were sampled from bleeding (BL) and non-bleeding (NBL) sites under camera-directed hysteroscopic guidance after Depo-provera injections. When compared with control biopsies, immunohistochemical examination revealed that 3 months of Depo-provera contraception reduced TF concentrations at the BL sites. However, there were ample EGFR and PR concentrations at BL and NBL sites. Moreover, there was a trend towards the appearance of pathologically enlarged blood vessels at the BL sites. The use of Western blotting revealed that after 3 months of Depo-provera, concentrations of both PRB and PRA isoforms were lower at BL versus NBL sites with decreased PRA concentrations attaining statistical significance. Separate sampling of endometrial BL and NBL sites as shown here for Depo-provera contraception could prove particularly useful in identifying local factors that determine the onset of bleeding during the more protracted time course of Norplant contraception. PMID- 11041231 TI - Bcl-2, Fas and caspase 3 expression in endometrium from levonorgestrel implant users with and without breakthrough bleeding. AB - Women using the progestin-only contraceptive Norplant often suffer from unpredictable bouts of breakthrough bleeding, which usually occurs from a thin atrophic endometrium. The role of cellular apoptosis in the endometrial response to Norplant has not been investigated. The aim of the present study was to use immunohistochemistry to produce semi-quantitative scores for expression of the apoptosis-related proteins Bcl-2, Fas and caspase 3 in endometrium from 16 controls and 42 women using Norplant with minimal or major breakthrough bleeding problems. The results showed no difference in endometrial immunostaining for any of the three proteins between Norplant users with and without breakthrough bleeding. There was also no evidence of endometrial endothelial cell immunostaining for any of the proteins. Bcl-2 was the only protein to show a cyclical pattern, with higher expression in the proliferative compared to secretory glands. All three proteins showed different expression levels in control functionalis versus basalis, with the survival protein Bcl-2 being higher in basalis, and the death receptor Fas and the proteolytic enzyme caspase 3 being higher in the functionalis. Overall, the results suggest that apoptosis is regulated differently in functionalis compared to basalis, and that atrophic Norplant-exposed endometrium appears more like functionalis than basalis with respect to expression of Fas and caspase 3. There was no evidence for a role for apoptosis in the mechanisms that underlie progestin-induced endometrial breakthrough bleeding. PMID- 11041232 TI - Morphological and functional changes in human endometrium following intrauterine levonorgestrel delivery. AB - The levonorgestrel releasing intrauterine system (LNG-IUS) provides a novel contraceptive method. Intrauterine LNG induces a dramatic transformation of the endometrium, characterized by extensive decidualization. This is associated with strong expression of local factors associated with decidualization, including prolactin receptor and insulin-like growth factor binding protein-1. A striking discovery was the down-regulation of oestrogen and progesterone receptors in all components of the endometrium after insertion of the LNG-IUS, with a gradual return between 6 and 12 months post-insertion. Preliminary findings suggest that androgen receptors are expressed during this time. Elevated leukocyte infiltrate is observed 1 month after insertion of the device, comprising large granular lymphocytes and macrophages. We examined a number of local mediators implicated in menstruation and breakthrough bleeding episodes. Expression of the chemokine interleukin-8 was enhanced after insertion of the device, with a notable decrease apparent 6 months post-insertion. Cyclooxygenase-2 was similarly strongly expressed in the first months after LNG-IUS insertion, in contrast to an initial suppression of prostaglandin dehydrogenase activity. By deduction it appears that higher local concentrations of prostaglandins are present in the initial period of local LNG exposure. Taken together these data suggest that in the first months following LNG-IUS insertion steroid receptor content is significantly decreased, resulting in the altered expression of many locally acting mediators which may be involved in breakthrough bleeding episodes. PMID- 11041233 TI - Insulin-like growth factors and insulin-like growth factor binding proteins in the endometrium. Effect of intrauterine levonorgestrel delivery. AB - Insulin-like growth factor (IGF) system is one of the growth factor systems that are believed to modulate steroid hormone actions in the endometrium through autocrine/paracrine mechanisms. IGF-I and IGF-II stimulate proliferation and differentiation, and maintain differentiated cell functions in several cell types in vitro. Endometrial stromal cells produce IGF-I and IGF-II as well as the high affinity IGF-binding proteins (IGFBP), whereas epithelial cells and, in a lesser amount, stromal cells contain cell membrane receptors for IGF. Oestrogen stimulates IGF-I gene expression, and IGF-II gene expression is associated with endometrial differentiation. The mRNA of six high affinity IGFBPs, which can modulate IGF actions, are expressed in human endometrium. The most abundant IGFBP in human endometrium is IGFBP-1, which is secreted by predecidualized/decidualized endometrial stromal cells in late secretory phase and during pregnancy. The primary negative regulator of IGFBP-1 production is insulin. IGFBP-1 competes with type I IGF receptor for binding of IGF in the endometrium and in cultured human trophoblastic cells. IGF-I mRNA is suppressed and mRNA encoding IGF-II and IGFBP-1 are consistently up-regulated in decidualized endometrium in women treated with the intrauterine levonorgestrel system (LNG-IUS). Strong cytoplasmic staining for IGFBP-1 was detected in decidualized endometrium in women using LNG-IUS for contraception or for endometrial protection during post-menopausal oestrogen replacement therapy. Simultaneously, oestrogen receptors were present, while progesterone receptors were hardly detectable in the endometrium by immunohistochemistry. The latter findings suggest that suppression of IGF-I action by IGFBP-1 may be one of the molecular mechanisms accounting for progestagenic and anti-oestrogenic effects of LNG-IUS in the endometrium. Consequently, examination of local IGF-I, IGF-II and IGFBP-1 expression might provide additional information when evaluating the effect of different progestins on the endometrium at the molecular level. PMID- 11041234 TI - Junctional barrier complexes undergo major alterations during the plasma membrane transformation of uterine epithelial cells. AB - Junctions in the plasma membrane of uterine epithelial cells as well as between these cells and their extracellular environment are examined in this review to see if a synthetic appreciation of their role can be gained from the disparate evidence presently available. Major changes in most junctional components are noted during early pregnancy and the role of progesterone and oestrogen in promoting these changes is examined. In particular it is noted that while tight junctions become deeper and morphologically 'tighter' towards the time of implantation, other basolateral junctional structures as well as their cytoskeletal associations are absent. These junctional alterations are part of the 'plasma membrane transformation' of early pregnancy and allow the conclusion that while paracellular permeability is reduced by the time of blastocyst attachment, the epithelial cells are paradoxically less firmly attached to each other, and to their extracellular environment. PMID- 11041235 TI - Endometrial epithelial integrity and subepithelial reticular fibre expression in progestin contraceptive acceptors. AB - Long-acting progestin contraceptives have been available in many countries for a number of years with a large number of women now using them. Although some improvements in delivery systems have been made, the major problem with progestin only contraceptives remains unpredictable endometrial breakthrough bleeding (BTB), which is responsible for more than 50% of drop-outs from this form of contraception. Using hysteroscopy, endometrial petechiae and ecchymoses are a common finding among Norplant users, although these features do not always correlate with BTB. It has been postulated that epithelial and subepithelial tissues may provide a barrier to BTB, as long as epithelial integrity is maintained. The aim of this pilot study is to explore structural changes in the endometrial surface epithelium, and subepithelial collagen III fibres. Endometrial biopsies from noresthisterone-enanthate (NetEn) users (n = 6) and controls (n = 6) were assessed using routine haematoxylin and eosin staining and immunohistochemical staining for cytokeratins 8, 18 and 19, and collagen III. A conventional silver impregnation method was also used to identify subepithelial collagen III fibres. Most of the Net-En tissues showed reduced surface epithelial cell height compared controls (P = 0.002). Cytokeratin staining as weaker (P = 0.04) and distributed evenly between basal and apical parts of the cell in Net-En tissue, compared to more apically in controls. Both immunohistochemical and conventional silver staining methods revealed that the subepithelial collagen III meshwork remained unchanged in Net-En compared to control endometrium. Both staining methods identified collagen fibres with equal sensitivity. In conclusion, atrophic changes remain the dominant appearance for progestin-exposed endometrium, with reduced cytokeratin staining, but apparently there is little change in subepithelial collagen III expression. PMID- 11041236 TI - Steroids and endometrial breakthrough bleeding: future directions for research. AB - Abnormal endometrial bleeding continues to be a significant problem for women using long-acting gestagens and not only reduces the use of these agents, thus restricting the options available to women, but also reduces the quality of their lives. This symposium addressed a range of basic and clinical studies that have tried over the past 3 years to define the problem. Two areas have been identified. The first is the structure of the blood vessels themselves and the second is the environment of the endometrium in relation to these vessels. This reflects a shift in research interest away from factors that simply control epithelial-mesenchyme interactions. The expression of angiogenic inhibitors and stimulators is being defined and the critical role of the matrix and the immunocompetent cells of the endometrium elucidated. The disappointing results of simple oestrogen supplementation were confirmed. Three broad areas of research were considered to be important for future studies. The first of these is not related to molecular and cellular function but rather to understanding better the attitudes of women to vaginal bleeding in the multicultural diverse situations in which this event occurs. It was recognized that a broader constituency was not interested in this problem with the increasing use of continuous combined regimes of hormone replacement therapy in the developed world. Studies that listen to and educate women about this problem are needed to ensure that these techniques are felt to be better owned by the women themselves. Molecular and cellular studies were identified that attempted to define the factors necessary for the development and maintenance of healthy, non-leaky vessels. These included studies of the pro- and anti-angiogenic agents expressed in endometrium and a definition of their interactions with matrix metalloproteinases in maintaining vessel integrity. Finally there was recognition of the need to understand how neutrophils traffic through the endometrium and to determine how they effect endometrial bleeding. PMID- 11041237 TI - TGF-beta/SMAD signaling and its involvement in tumor progression. AB - Cytokines of the transforming growth factor-beta (TGF-beta) superfamily are multifunctional peptides that regulate growth and differentiation of various types of cells. Members of the TGF-beta superfamily bind to type 11 and type I serine/threonine kinase receptors, which mediate intracellular signals through SMAD proteins. Of 3 subtypes of SMADs, receptor-regulated SMADs are phosphorylated by the serine/threonine kinase receptors, form complexes with common-mediator SMAD, and move into the nucleus, where they act as components of transcription factor complexes. Abnormalities of the TGF-beta receptors and SMADs have been detected in various tumors, including colorectal cancers and pancreatic cancers. Inhibitory SMADs and transcriptional co-repressors, including c-Ski and SnoN, repress the TGF-beta/SMAD signaling. Perturbation of the TGF-beta/SMAD signaling pathway may result in progression of tumors through resistance of the cells to the growth inhibition induced by TGF-beta. PMID- 11041238 TI - High throughput detection of drug-metabolizing enzyme polymorphisms by allele specific fluorogenic 5' nuclease chain reaction assay. AB - We have developed an allele-specific fluorogenic 5' nuclease chain reaction assay for detecting polymorphisms in the following human drug-metabolizing enzyme genes: CYP2C9 (CYP2C9*2 and *3), CYP2C19 (CYP2C19*2 and *3), CYP2D6 (CYP2D6*4, *10, *14, *18, and *21(C8)), N-acetyltransferase 2 (NAT2*5B, *6A, and *7B), thiopurine methyltransferase (TPMT*3C), and aldehyde dehydrogenase2 (ALDH2*2). This method is a marriage of two emerging technologies, the use of allele specific amplification primers for target DNA and hybridization of the TaqMan probe. The TaqMan probe is labeled with both a fluorescent reporter dye and a quencher dye. Genotypes are separated according to the different threshold cycles of the wild-type and mutant primers. All assays are performed using a single thermocycling protocol. This genotyping method is rapid and highly sensitive and yields a high throughput. It could be applied toward automated large-scale genotyping. PMID- 11041239 TI - Effect of murine kidney extracts on the proliferation of hematopoietic progenitor cells in human umbilical cord blood. AB - We examined the effect of murine kidney extract (MKE) on the clonal growth of highly purified CD34+ hematopoietic progenitor cells from human umbilical cord blood. MKE did not affect the total number of colonies of erythroid burst-forming units (BFU-E), granulocyte-macrophage colony-forming units (CFU-GM) or granulocyte-erythroid-macrophage-megakaryocyte colony-forming units (CFU-Mix/CFU GEMM) in a methylcellulose culture with exogenous recombinant human granulocyte colony-stimulating factor, granulocytemacrophage colony-stimulating factor, interleukin-3, stem cell factor and erythropoietin. MKE significantly increased the proportion of BFU-E- or CFU-Mix-derived colonies, and suppressed the formation CFU-GM-derived colonies depending on the MKE dose. However, because of an increase in small megakaryocyte colonies derived from mature CFU-Meg MKE increased by approximately 40% the growth of megakaryocyte colony-forming units (CFU-Meg) in plasma clot culture stimulated by recombinant human thrombopoietin. Also MKE promoted an increase in hyperploid megakaryocytes, suggesting that the active factor(s) in MKE acts on the mature CFU-Meg and promotes the maturation of megakaryocytes. Gel-filtration high performance liquid chromatography of MKE showed that the promoting factor(s) in MKE was approximately 45 kDa. These results indicate that the factor(s) detected in MKE influence human hematopoiesis in vitro, especially thrombopoiesis. PMID- 11041240 TI - Bile acids in porcine fetal bile. AB - A study of the biliary bile acid composition in porcine fetus compared with that of the adult pig is described. Biles, collected during gestation (weeks 4, 15 to 17 and at birth), aged six months and two years old, were analyzed by gas-liquid chromatography and capillary GC-MS. Bile acids were separated into different conjugate groups by chromatography on the lipophilic anion exchange gel, piperidinohydroxypropyl Sephadex LH-20. All and one fourth of the total bile acids in the bile of weeks 4 and 15 of gestation, respectively, were present as unconjugated form, however, only a trace of unconjugated bile acids was present in bile of late gestation, the young and the adult pigs. The ratio of glycine/taurine (G/T) conjugates in the conjugated fraction of the fetal bile at 15 weeks gestation was less than 1, which markedly contrasted with the conjugation pattern for adult bile where the ratio of G/T conjugates was approximately more than 9. The predominant acids identified in porcine fetal bile of the 4 weeks gestation were cholic acid (3alpha,7alpha,12alpha-trihydroxy-5beta chola n-24-oic acid) and chenodeoxycholic acid (3alpha,7alpha -dihydroxy-5beta cholan-24-oic acid). However, cholic acid in late gestation, young, and adult bile was the smallest component, whereas chenodeoxycholic acid was still the major constituent of these biles. The presence of small but valuable amounts of allocholic acid (3alpha,7alpha,12alpha-trihydroxy-5alpha-chol an-24-oic acid) and cholic acid in early gestation suggested the presence of 12alpha-hydroxylase activity of steroid nucleus in fetal liver. Considerable amounts of glycine conjugated hyodeoxycholic acid were found in the bile of the gestation periods, suggesting the placental transfer of this bile acid from maternal circulation. PMID- 11041241 TI - NMR structure of ribonuclease HI from Escherichia coli. AB - The solution structure of ribonuclease HI (RNase HI) from Escherichia coli (E. coli), a protein of 155 residues, was determined. Three-dimensional nuclear Overhauser enhancement spectroscopy (NOESY) was used to obtain 1,424 distance constraints between individually assigned polypeptide chain hydrogen atoms. Supplemental geometric constraints of 90phi angles and 12chi1 angles, and the distance constraints of 66 hydrogen bonds were experimentally derived. Using the DADAS90 program that calculates structures in dihedral angle space, 15 structures satisfying almost all constraints were obtained. The average root mean square deviation (RMSD) from the mean structure was 0.75 A for backbone atoms. The RMSD for backbone atoms between the representative NMR structure with the smallest constraint violation and crystal structures was within 1.2 A. Although the NMR and crystal structures thus resemble one another, a significant discrepancy was observed in a region termed 'basic protrusion.' The discrepancy observed in NMR experiments is explained by fluctuation in this region. PMID- 11041242 TI - Role of reactive oxygen species in gallic acid-induced apoptosis. AB - We earlier demonstrated that gallic acid (3,4,5-trihydroxybenzoic acid) induced apoptosis in promyelocytic leukemia HL-60RG cells, which was inhibited by catalase and intracellular Ca2+ chelator. In this study, we further studied the involvement of reactive oxygen species (ROS) and intracellular Ca2+ in gallic acid-induced apoptosis. The enhancement of intracellular ROS in HL-60RG cells was detected dose-dependently as early as 5 min after stimulation with gallic acid by using 5,6-carboxy-2',7'-dichlorofluorescin diacetate (DCFH-DA). Further studies that used various antioxidants and ROS scavengers showed that the intracellular peroxide level was well correlated with the potency to induce apoptosis and that the increased intracellular peroxides after gallic acid treatment seemed likely to result from the influx of H2O2 derived from superoxide which were generated extracellularly. In addition, gallic acid, HX/XO, and H2O2-induced apoptosis was completely inhibited by pretreatment with intracellular Ca2+ chelator 1,2-bis(2 aminophenoxyethane)-N,N,N'-tetraacetic acid tetrakis (acetoxymethyl ester) (BAPTA AM), but increase of intracellular peroxide levels by gallic acid were suppressed only slightly. It is suggested that intracellular ROS induced by gallic acid plays an important role in eliciting an early signal in apoptosis. Especially, H2O, which is derived from superoxide anion generated extracellularly may increase intracellular Ca2+ levels or cooperate with intracellular Ca2+, thus resulting in apoptosis induction. PMID- 11041243 TI - Detection of kinases that phosphorylate 14-3-3 binding sites of Raf-1 using in situ gel kinase assay. AB - Raf-1 is a serine/threonine protein kinase that plays a critical role in mitogenic signal transduction. Raf-1 activation requires 14-3-3 binding to Raf-1 as an essential step. This binding is regulated through phosphorylation of Ser259 and Ser621 of Raf-1, each constituting part of the consensus motif for the binding of Raf-1 to 14-3-3. However, Raf-1 kinase kinase(s) that phosphorylates these sites remains unknown. In this report, we detected Raf-1 kinase kinase activity using recombinant glutathione-S-transferase-Raf-1 fusion proteins as substrate of in situ gel kinase assay. Ser259 was phosphorylated by a kinase with a molecular weight of 90 kDa, which was suggested to be Rsk judging from the molecular size, the time course of activation after EGF stimulation and the elution pattern from an anion-exchange column. The Raf-1 fragment containing Ser621 was phosphorylated by kinases with molecular weights of 85, 60, 50 and 48 kDa but not by the kinase that phosphorylates Ser259. These results suggest that although Ser259 and Ser621 lie in the same amino acid sequence motif for 14-3-3 binding, these two regulatory sites for this binding are phosphorylated by different protein kinases. PMID- 11041244 TI - Detection and characterization of a type IIA secretory phospholipase A2 inhibitory protein in human amniotic fluid. AB - Two types of phospholipase A2 (PLA2) inhibitory protein (PLIP-I, PLIP-II) were detected and isolated from human amniotic fluid by Sephacryl S300 gel filtration chromatography. The lower molecular weight-fraction (PLIP-II) was further purified by Sephadex G75 gel filtration and analyzed by SDS-PAGE. Its molecular weight was estimated to be approximately 18 kDa, and it was sensitive to heat treatment. Inhibition of phospholipase A2 (sPLA2 type IIA) by PLIP-II occurred in a dose-dependent manner (IC50 about 0.82/microm), and the effect was stronger on sPLA2 IIA than on pancreatic sPLA2 (IC50 about 3.11 microL). The ratio of the inhibitions of the sPLA2 IIA by PLIP-II remained consistent over an entire range of substrate concentrations. Furthermore, addition of excess Ca2+ at concentrations of up to 10 mm did not antagonize the inhibitory activity of PLIP II. PMID- 11041245 TI - Characterization of bovine and human lactoferrins as glycyrrhizin-binding proteins and their phosphorylation in vitro by casein kinase II. AB - The binding ability of bovine and human lactoferrins (bLF and hLF; LFs) to a glycyrrhizin (GL)-affinity column and their phosphorylation by casein kinase II (CK-II) in vitro were biochemically investigated. It was found that (i) both bLF and hLF are GL-binding proteins; (ii) purified both proteins function as phosphate acceptors of CK-II; and (iii) this phosphorylation is completely inhibited by two polyphenol-containing anti-oxidant compounds (quercetin and epigallocatechin gallate) at I microm, whereas a glycyrrhetinic acid derivative (oGA) inhibits it at one tenth the concentration of GL. The DNA-binding affinity of hLF was reduced by GL in a dose dependent manner. However, no significant effect of the CK-II-mediated hLF phosphorylation on its DNA-binding affinity was detected. These results suggest that the GL-induced inhibition of the DNA-binding affinity and the CK-II-mediated phosphorylation of hLF may be closely correlated with the anti-inflammatory effect of GL in the human body. PMID- 11041246 TI - Reaction of various lectins to mucin derived from the different layers of rat gastric mucosa: comparison of enzyme-linked lectin binding assay with lectin histochemistry. AB - The purpose of this study was to demonstrate that the histochemical staining reactivities of lectins in rat stomach actually represent the gastric mucins, and to estimate the utility of the lectins for mucin histochemistry. In this paper, the lectin histochemistry was compared with an enzyme-linked lectin binding assay (ELLA) of the mucins derived from distinct regions and layers of the Sprague Dawley rat stomach and it was examined to determine the definite binding and problematic binding of the conventional lectin. Among the 10 different biotinylated lectins, Canavalia ensiformis (ConA), Griffonia simplicifolia II (GS II), Triticuin vulgaris (WGA), Ricinus communis I (RCA-I), Arachis hypogaea (PNA), Glycine max (SBA), Dolichos biflorus (DBA), Ulex europaeus I (UEA-I), Sambucus nigra (SNA) and Maackia amurensis II (MAL-II), examined in this study, GS-II, SBA, DBA, UEA-I, SNA and MAL-II bound clearly to the mucin of distinct regions and layers of the Sprague-Dawley rat stomach in agreement with the results of ELLA. Namely GS-II lectins preferentially bound to the mucin in the mucous neck cells of the corpus area. SBA and DBA clearly recognized the mucin in the covering epithelial mucous cells in the corpus and antral area. UEA-I was widely bound to all the mucin present in both the corpus and antrum. On the other hand, SNA and MAL-II could not react with the mucin obtained from the gastric mucosa but was specifically bound to the mucin purified from the mucous gel layer. These results suggested that the lectins described above are useful histochemical tools to recognize the mucus present in the different regions and layers of Sprague-Dawley rat gastric mucosa. PMID- 11041247 TI - Fucoidan is the active component of fucus vesiculosus that promotes contraction of fibroblast-populated collagen gels. AB - The fibroblast-populated collagen gel culture method has been evaluated as a dermal model of wound contraction and granulation in tissues during the wound healing process and as an in vitro model of dermal tissue. We previously reported that an extract of Fucus vesiculosus promoted fibroblast-populated collagen gel contraction and that the promotion of the gel contraction was due to the increased expression of integrin alpha2beta1 on the surface of the fibroblasts. In this study, we investigated the active component of the extract of this alga using extraction and fractionation techniques. Water extraction of the alga was followed by precipitation with excess ethanol and then gel filtration with the boundary molecular weight of 30,000. The high molecular weight fraction obtained from gel filtration was fractionated by ion exchange chromatography on diethylaminoethyl cellulose column to give active fractions that have more polar properties. These polar, high molecular weight fractions which contained molecules with fucose and sulfate groups showed significant gel contraction promoting activity and integrin expression-enhancing activity, and were estimated to be the sulfated-polysaccharide fucoidan. Commercially available fucoidan showed similar activities to the above-described fraction of this alga. Although it remains necessary to precisely identify the specific active component, the above results indicate that fucoidan is the active component which promotes collagen gel contraction, and also indicate the possibility that it dose so by enhancing the integrin alpha2beta1 expression. PMID- 11041248 TI - Evidence for the AT1 subtype of the angiotensin II receptor in the rat submandibular gland. AB - We have characterized angiotensin II (Ang II) receptor subtypes on rat submandibular gland membranes using a radioligand binding assay. [3H]Ang II binding to the membrane fractions exhibited both high (Kd =0.08 nm, Bmax =2.19 fmol/mg protein) and low (Kd =4.19 nm, Bmax = 13.7 fmol/mg protein) affinity. Ang 11, Ang III and saralasin completely displaced the [3H]Ang II binding, whereas CV 11974, an AT1 receptor antagonist and PD123319, an AT2 receptor antagonist maximally displaced up to approximately 87 and 13% of the total binding, respectively. [3H]DuP753 binding to the membrane fractions exhibited a single population of binding site with a Kd of 4.22 nM and Bmax of 3.77 pmol/mg protein. Ang II, Ang III and CV-11974 completely displaced the [3H]DuP753 binding with slope factors near unity, but PD123319 did not. These findings suggest that rat submandibular gland membranes contain predominantly the AT1 receptor subtype. PMID- 11041249 TI - Improvement in the histopathology of hearts from cardiomyopathic BIO TO-2 hamsters following long-term administration of amlodipine and cilnidipine. AB - The effect of long-term administration of amlodipine and cilnidipine was examined on the histopathology and 1,4-dihydropyridine (DHP) calcium channel antagonist receptors in the left ventricle of BIO TO-2 hamsters, a model of dilated cardiomyopathy (DCM). Oral administration of amlodipine (3 and 10 mg/kg/d, 19 weeks) in 7 week-old BIO TO-2 hamsters produced a significant reduction in calcium deposition and necrosis with little change in the cavity area and fibrosis. A reduction of calcium deposition and necrosis in the myocardium of BIO TO-2 hamsters was also seen following similar administration of cilnidipine (10 mg/kg/d). The long-term administration of amlodipine (3 and 10 mg/kg/d) caused a significant increase (36.6 and 21.7%, respectively) in the Bmax for specific (+) [3H]PN 200-110 binding in the myocardium from BIO TO-2 hamsters, compared with that in control hamsters. In conclusion, the present study has shown that long term administration of amlodipine and cilnidipine improves calcium deposition and necrosis in the myocardium from BIO TO-2 hamsters. Thus, these data suggest that both agents may be effective pharmacological treatments of DCM. PMID- 11041250 TI - Anti-Inflammatory properties of a lipid fraction obtained from Sideritis javalambrensis. AB - A lipid fraction obtained by activity-guided fractionation from the hexane extract of Sideritis javalambrensis was evaluated for anti-inflammatory activity. This fraction significantly inhibited paw oedema induced by carrageenan as well as ear oedema induced by 12-O-tetradecanoylphorbol 13-acetate (TPA) in mice after oral or topical administration, respectively. Quantitation of the specific marker myeloperoxidase (MPO) demonstrated that its topical anti-inflammatory activity was associated with reduction in cell infiltration into inflamed tissues. The lipid fraction significantly decreased leukocyte granular enzyme release (beta glucuronidase), but failed to inhibit superoxide generation. Histamine release from mast cells was also suppressed in a concentration-dependent manner. In addition, non-toxic concentrations of this fraction reduced nitric oxide (NO) generation in lipopolysaccharide (LPS)-treated J774 macrophages. Taken together, these results suggest that the lipid fraction exerts in vivo anti-inflammatory activity with the partial contribution of inhibitory actions on some inflammatory responses. PMID- 11041251 TI - Extracellular signal regulated protein kinase and c-jun N-terminal kinase are involved in ml muscarinic receptor-enhanced interleukin-2 production pathway in Jurkat cells. AB - We have previously shown that m1 and m2 muscarinic receptors were expressed on human peripheral blood lymphocytes (hPBL) and that pre-stimulation of these receptors enhanced phytohemagglutinin (PHA)-induced interleukin-2 (IL-2) production. Possible intracellular signal pathways of muscarinic receptors to regulate IL-2 production were examined in human T cell line Jurkat cells. Pretreatment of the cells with muscarinic receptor agonist, oxotremorine M (Oxo M), enhanced IL-2 production induced by phorbol 12-myristate 13-acetate (PMA)/A23187, while Oxo-M by itself did not affect IL-2 production. The enhancement of IL-2 production by Oxo-M was inhibited by 4-diphenylacetoxy-N methylpiperidine methiodide (4-DAMP) an ml/m3 receptor antagonist. When the cells were pretreated with AF-DX116, an m2 antagonist, the IL-2 production enhanced by Oxo-M was further stimulated. Reverse transcription-polymerase chain reaction (RT PCR) revealed that ml and m2 muscarinic receptors exist on Jurkat cells. The stimulation of ml receptors enhanced the PMA/A23187-induced binding activity to AP-1 consensus sequences in IL-2 promoter and production of c-Fos and c-Jun protein. The stimulation of ml receptors did not modify the DNA binding of NF kappaB, NF-AT or Oct-1. When ml receptors were stimulated, activities of mitogen activated protein kinase (MAPK)/extracellular signal regulated protein kinase (ERK) and c-Jun N-terminal kinase (JNK) were increased, while p38 MAPK was not affected. Incubation with Oxo-M induced a transient increase in [Ca2+]i, which was abolished by pretreatment with 4-DAMP. Treatment with cyclosporin A markedly decreased the PMA/A23187-induced IL-2 promoter activity. This treatment, however, did not affect the enhancement of the promoter activity induced by ml receptor stimulation. The results suggest that transcription factor AP-1 is involved in the ml receptor-mediated enhancement of IL-2 transcript in Jurkat cells, and that pathways via MAPK/ERK and JNK, but not via p38 MAPK, are involved in the ml receptor-mediated enhancement of IL-2 promoter activity. PMID- 11041252 TI - Effect of biphenyl dimethyl dicarboxylate on the cellular and nonspecific immunotoxicity by ethanol in mice. AB - Biphenyl dimethyl dicarboxylate (PMC) has been reported to protect against chronic ethanol toxicity. The present study was conducted to evaluate whether PMC might be accompanied by a reduction of ethanol-induced cellular immunotoxicity. PMC at a dose of 6 mg/kg was orally administered to ICR mice daily for 28 consecutive days, and normal mice were given vehicle. Mice treated with ethanol were given free access to 20% w/v ethanol instead of water. Mice were immunized and challenged with sheep red blood cells (SRBC). Delayed-type hypersensitivity (DTH) reaction to SRBC was increased to normal levels by the combination of PMC and ethanol, compared with the treatment of ethanol alone. Splenic CD4+ cells were also greatly enhanced by PMC treatment as compared with the treatment of ethanol alone. In the case of CD8+ cells, however, a slight reduction was observed by the PMC or ethanol treatment. The natural killer (NK) cell and phagocytic activity used for evaluation of nonspecific immunocompetence were significantly augmented in PMC plus ethanol-treated mice when compared with the treatment of ethanol alone. The number of peripheral leukocytes was significantly decreased by the treatment of ethanol alone, then also restored to normal levels by PMC treatment. These findings indicate that cellular immunotoxicity caused by ethanol consumption is significantly restored or prevented by PMC treatment. PMID- 11041253 TI - 72-kDa stress protein (hsp72) induced by administration of dimethylarsinic acid to mice accumulates in alveolar flat cells of lung, a target organ for arsenic carcinogenesis. AB - Our previous studies have demonstrated that the oral administration of dimethylarsinic acid (DMA), a main metabolite of inorganic arsenics in mammals, in mice causes DNA damage in the lung as well as the promotion and progression of lung- and skin-tumorigenesis. Moreover, we indicated that 72-kDa stress protein (Hsp72) was induced in cultured human pulmonary (L-132) cells by exposure to DMA and was accumulated specifically in the cell nuclei. The present in vivo study reveals the induction of Hsp72 by intraperitoneal administration of DMA to A/J mice used previously as an animal model of dimethylarsenic-induced lung tumorigenesis. The Hsp72 was observed in the lung, a target organ for arsenic carcinogenesis in human, and in the kidney as well, but not in the liver and spleen. By immunohistochemical analysis, the Hsp72 in lungs was exhibited to exist in the nuclei of alveolar flat cells, including capillary endothelial cells, which were previously found to increase the clumping of heterochromatin, an early morphological change in the developmental process of pulmonary carcinomas, after administration of DMA to mice. These in vivo observations suggest that the increase and accumulation of Hsp72 by administration of DMA to mice may occur specifically in target organs for arsenic carcinogenesis. PMID- 11041254 TI - Constituents from the stem and root of Aristolochia kaempferi. AB - Three new phenanthrene derivatives, aristoliukine-C, aristofolin-E and aristolochic acid-Ia methyl ester, and one new sesquiterpene, madolin-P, together with 58 known compounds were isolated from the stem and root of Aristolochia kaempferi. The structures of these compounds were determined by spectral analysis. The cytotoxicity and antiplatelet activity of the isolated compounds are also discussed. PMID- 11041255 TI - The effects of Pva/chitosan/fibroin (PCF)-blended spongy sheets on wound healing in rats. AB - The effects of poly vinyl alcohol (PVA)/Chitosan/Fibroin (PCF)-blended sponge on wound healing in rats were investigated. We excised the skin of a rat, including the dermis, approximately 2 x 2 cm in size. The wound was covered with PCF blended spongy sheets. The spongy sheets absorbed the exudate, and gained flexibility and softness. Histopathological inspection of the wound 12 d later showed an increase of vascular ingrowth and the absence of inflammatory cells. Regeneration of the skin around the wound was faster than that of the control. We also tested wound healing effects of PVA, Chitosan and Fibroin, alone or in various combinations. Wound healing was accelerated in the order of PCF-blended sponge>Chitosan/Fibroin (CF)-blended sponge>Fibroin (F) sponge > PVA/Chitosan blended sponge (PC) > Chitosan (C) sponge. PMID- 11041256 TI - Effect of vehicle properties on skin penetration of emedastine. AB - We investigated sorption and permeation of emedastine with 11 different vehicles, composed of single or binary solvents, in excised rat skin. In the sorption study, partition parameters of the drug with each vehicle were obtained by dividing the drug amount in skin at equilibrium by its donor concentration. When the logarithm of the partition parameters for the stratum corneum/vehicle partitioning (Ks') was plotted against the dielectric constants of the vehicles, a bi-linear relationship was obtained. The skin flux of emedastine largely differ among the vehicles. A quasi-steady-state flux of emedastine exhibited a good linear relationship with Ks', except for ethanol (EtOH)/isopropyl myristate (IPM) systems, indicating that the partitioning process is critical in determining the permeation rate. Delineation of the EtOH/IPM systems would be due to an increase in the diffusivity of the drug in the stratum corneum, as indicated by the analysis using a two-layer diffusion model. Thus, differential evaluation of partitioning and diffusion processes by both sorption and permeation studies would give further insights into the effects of vehicles on skin permeation of drugs. PMID- 11041257 TI - Application of fibroin in controlled release tablets containing theophylline. AB - The applicability of fibroin, a major silk protein, to controlled release type dosage tablets was investigated in vitro and in vivo. Fibroin tablets containing theophylline were easily prepared by a direct compression method without additives. Five types of fibroin tablets with the same surface area and different amounts of theophylline were prepared. In an in vitro drug study, the drug release from the tablets was not affected by the pH of the release medium. The greater the fibroin content in the tablets, the lower the percentage released at time t. The Higuchi plots of the release data showed a linear release profile, indicating that the drug release from the fibroin tablets was diffusion controlled through the matrix. Theophylline powder or a TF-41 tablet (theophylline: fibroin=4 : 1), or a commercial tablet, Uniphyl (once-a-day type) was administered to 5 healthy volunteers. The areas under the saliva theophylline concentration-time curve (AUC) of Uniphyl and TF-41 to that of powder were 85% and 70%, respectively (fasted). Conversely, the mean residence time and mean absorption time of TF-41 were long compared to Uniphyl (fasted). Therefore, the reduction of bioavailability in TF-41 was due to the delayed release from the tablets in vivo. Taken after a meal, the AUCs of TF-41 and Uniphyl increased and the absorption was completed. This suggested that the drug release from TF-41 may increase due to the stimulation of food on TF-41 itself and due to movement of the gastro-intestinal tract. In conclusion, fibroin could be used as the matrix in controlled-release tablets. PMID- 11041258 TI - Bactericidal activity of lemon juice and lemon derivatives against Vibrio cholerae. AB - Food products can be possible vectors of the agent responsible for cholera epidemics, because some of these products allow Vibrio cholerae O1 to develop to concentrations above the dangerous level. This study deals with the behaviour of essential oils, natural and concentrated lemon juice and fresh and dehydrated lemon peel against V. cholerae O1 biotype Eltor serotype Inaba tox+. Our aim was to evaluate whether these products, used at different dilutions, exhibit bactericidal or bacteriostatic activity against the microorganism, when present at concentrations of 10(2), 10(4), 10(6) and 10(8) colony forming units (CFU) ml( 1), and after different exposure times. 10(8) CFU ml(-1) was considered an infectious dose. Concentrated lemon juice and essential oils inhibited V. cholerae completely at all studied dilutions and exposure times. Fresh lemon peel and dehydrated lemon peel partially inhibited growth of V. cholerae. Freshly squeezed lemon juice, diluted to 10(-2), showed complete inhibition of V. cholerae at a concentration of 10(8) CFU ml(-1) after 5 min of exposure time; a dilution of 2 x 10(-3) produced inhibition after 15 min and a dilution of 10(-3) after 30 min. It can be concluded that lemon, a natural product which is easily obtained, acts as a biocide against V. cholerae, and is, therefore, an efficient decontaminant, harmless to humans. PMID- 11041259 TI - Tissue-Specific expression of rat kininogen mRNAs. AB - To characterize the local kallikrein-kinin system, tissue distribution of mRNAs for kininogens, precursor proteins of kinins, such as high-molecular-weight (H-), low-molecular-weight (L-) and T-kininogens, were studied in the rat by means of reverse-transcription polymerase chain reaction (RT-PCR) using a fluorophore Cy5 labeled 5'-primer. High levels of these three kininogen mRNAs were present in the liver. Relatively high levels of H-kininogen mRNA were also detected in the skin, lung, kidney, and testis in a descending order, whereas L-kininogen mRNA was detectable in the lung and brain, but not in the kidney, skin, testis, heart, adrenal gland, or skeletal muscle. T-Kininogen mRNA was present in these tissues, except for skeletal muscle. These findings suggest that the expression of each kininogen mRNA is regulated by the tissue-dependent mechanisms which is closely associated with functions of the kallikrein-kinin system in the tissue. PMID- 11041260 TI - Hepatoprotection by human epidermal growth factor (hEGF) against experimental hepatitis induced by D-galactosamine (D-galN) or D-GalN/lipopolysaccharide. AB - The hepatoprotecive effects of recombinant human epidermal growth factor (hEGF) on chemically and immunologically induced experimental liver injury models were examined. The hEGF clearly decreased serum transaminase levels in D-galactosamine (D-GalN) and D-GalN/lipopolysaccharide (LPS)-induced liver injury models under sub-lethal conditions. However, it has not significantly changed either serum or in vitro tumor necrosis factor (TNF)-alpha production or in vitro nitric oxide (NO) production, suggesting that the hepatoprotection by EGF is not mediated by inhibiting these pathological mediators produced in D-GalN and D-GalN/LPS-induced liver injury. PMID- 11041261 TI - Antimutagenic activity of 5alpha-cholest-7-en-3beta-ol, a new component from the starfish asterina pectinifera. AB - From the butanol fraction of the starfish Asterina pectinifera Muler et Troschel (Asteriidae), we have isolated a new component, 5alpha-cholest-7-en-3beta-ol. Its antigenotoxic and antimutagenic activities were examined by the SOS chromotest with Escherichia coli PQ37 and by Ames test with Salmonella typhimurium TA1538, respectively. 5alpha-Cholest-7-en-3beta-ol showed potent antigenotoxic activity against the mutagens, both MNNG and NQO. For 100% of antigenotoxicity, the concentration of the compound applied against MNNG and NQO were 10 microg and 5 microg per reaction tube, respectively. Its antimutagenic activity with S. typhimurium TA1538 against the mutagen MNNG was very effective. When its concentrations were varied from 1 up to 10 microg dose per plate, the inhibition ratio of revertant CFU of TA1538 per plate was increased accordingly, from 25.2 to 99.2%. These results suggest that 5alpha-cholest-7-en-3beta-ol possesses antigenotoxic and antimutagenic activity and might be useful as a chemopreventive agent. PMID- 11041262 TI - Screening of crude drugs for influence on amylase activity and postprandial blood glucose in mouse plasma. AB - Screening of crude drugs for inhibitory activity on alpha-amylase in mouse plasma was performed. Hot water extracts and ethanolic extracts of Arecae Semen, Ephedrae Herba, Malloti Cortex, Rhei Rhizoma and ethanolic extract of Moutan Cortex inhibited enzyme activity in isolated mouse plasma strongly and dose dependently. However, the effects of these 9 extracts were not observed in the plasma when they were administered intraperitoneally or orally. Ethanolic extract of Arecae Semen also showed a depressive effect on elevation of postprandial blood glucose. PMID- 11041263 TI - Effect of l-menthol-ethanol-water system on the systemic absorption of flurbiprofen after repeated topical applications in rabbits. AB - The effect of the l-menthol-ethanol-water system (MEW system), a skin penetration enhancer, on the systemic absorption of flurbiprofen (FP) after repeated topical applications was investigated. FP (1%) gel containing ethanol (25%) and l-menthol (3%) as penetration enhancers was applied to rabbit dorsal skin and the in vivo absorption rate of FP was compared with the in vitro penetration rate through excised skin. In vivo absorption rate of FP was initially high and decreased with time to a value approximately equal to the in vitro rate. The remaining FP in the gel 6 h after the application was 60% of the initial loading and the systemic bioavailability over the 6 h application was about 10%, suggesting that the rest (30%) had accumulated in the skin tissues. The gel was applied for 6 h on the same site or on a new site after the first 6 h-application to learn the effect of repeated applications on FP absorption. The maximum FP concentration after the second application on the virgin skin was slightly higher than that after the first application, as expected in a typical pharmacokinetic process. In contrast, the same site application induced remarkably lower plasma concentration and area under the curve (AUC). A drug-free gel was also utilized to evaluate the effects of the enhancer system. Pretreatment of the drug-free gel on the same site also decreased the FP absorption, whereas post-treatment increased the plasma level of FP, in spite of the removal of the drug gel. These phenomena could be explained by ethanol in the MEW system acting a local irritant and a drug carrier. PMID- 11041264 TI - Bidirectional effects of hypergravity on the cell growth and differentiated functions of osteoblast-like ROS17/2.8 cells. AB - A low level of hypergravity (1.5-2.0 G) stimulated the proliferation of ROS17/2.8 cells, whereas it inhibited the differentiated functions of alkaline phosphatase activity and osteocalcin synthesis. These results were just the opposite of our results obtained when the cells were exposed to a high level of hypergravity (40 80 G): inhibition of cell growth and the stimulation of the differentiated functions. The direction of change in the cAMP contents of the cells was also reversed, with a low level of hypergravity causing a decrease in the cAMP content and a high level of hypergravity an increase in it. Therefore, bidirectional effects of hypergravity on the growth and differentiated functions exist in ROS17/2.8 cells according to the magnitude of the hypergravity. PMID- 11041265 TI - Marked therapeutic effect of dimyristoylphosphatidylcholine liposomes on carcinoma mice model in vivo. AB - Significantly prolonged survival rate was obtained for the first time using carcinoma mice models after the administration of single-component liposomes of dimyristoylphosphatidylcholine (DMPC) without any drug. An increase in lymphocytes under optical microscope was observed without any increase in the neutrophils count, suggesting that the DMPC liposomes might inhibit the tumor growth as well as increase in lymphocytes in vivo. PMID- 11041266 TI - Development of a Simple Cell Invasion Assay System. AB - Cell invasion assay is important for studying various biological events, such as inflammation, cancer metastasis, and angiogenesis. In this study, we developed a simple method for the quantification of cell invasion by using a culture insert with fluorescence blocking micropore membrane (FBM). Fluorescence labeled cells were simply added to a culture insert with a 8 micrompore FBM precoated with Matrigel and incubated for an appropriate duration. Then, the FBM was examined under a fluorescence microscope to count the invaded cell number. By this method, accurate invasion assay is easily performed without the steps of fixation and staining of cells and removal of cells which do not invade. PMID- 11041267 TI - Adenosine A2A receptor gene expression in the normal striatum and after 6-OH dopamine lesion. AB - Adenosine A2A receptors are present on enkephalinergic medium sized striatal neurons in the rat and have an important function in the modulation of striatal output. In order to establish more accurately whether adenosine transmission is a generalized phenomenon in mammalian striatum we compared the A2A R expression in the mouse, rat, cat and human striatum. Secondly we compared the modulation of enkephalin gene expression and A2A receptor gene expression in rat striatal neurons after 6-OH-dopamine lesion of the substantia nigra. Hybridization histochemistry was performed with a 35S-labelled radioactive oligonucleotide probe. The results showed high expression of A2A adenosine receptor genes only in the medium-sized cells of the striatum in all examined species. In the rat striatum, expression of A2A receptors was not significantly altered after lesion of the dopaminergic pathways with 6-OH-dopamine even though enkephalin gene expression was up-regulated. The absence of a change in A2A receptor gene expression after 6-OH-dopamine treatment speaks against a dependency on dopaminergic innervation. The maintained inhibitory function of A2A R on motor activity in spite of dopamine depletion could be partly responsible for the depression of locomotor activity observed in basal ganglia disorders such as Parkinson's disease. PMID- 11041268 TI - Intraaccumbal mecamylamine infusion does not affect dopamine output in the nucleus accumbens of chronically nicotine-treated rats. AB - Recently we have shown that the nicotinic receptor (nAChR) antagonist mecamylamine both when administered systemically and locally into the ventral tegmental area (VTA) to chronically nicotine-treated rats reduces dopamine (DA) output in the nucleus accumbens (NAC) and elicits behavioral withdrawal signs. However, the putative contributory role of nAChRs in the NAC in mediating these effects of systemic mecamylamine has not been clarified. Therefore, we here investigated the effect on extracellular levels of DA in the NAC of local intraaccumbal administration of mecamylamine to chronically nicotine-treated rats and its putative behavioral correlates. In these experiments local application of mecamylamine into the NAC, in a concentration that increased NAC DA levels in control rats, did not affect DA output or behavior in the nicotine-treated animals. These results provide further support for the contention that nAChRs in the VTA, but not in the NAC, are of major importance for the mesolimbic DA reduction and associated behavioral signs in nicotine withdrawal. PMID- 11041269 TI - Sequential changes of cholinergic and dopaminergic receptors in brains after 6 hydroxydopamine lesions of the medial forebrain bundle in rats. AB - We studied sequential changes in muscarinic cholinergic receptors, high-affinity choline uptake sites and dopamine D2 receptors in the brain after 6 hydroxydopamine lesions of the medial forebrain bundle in rats. The animals were unilaterally lesioned in the medial forebrain bundle and the brains were analyzed at 1, 2, 4 and 8 weeks postlesion. [3H]Quinuclidinylbenzilate (QNB), [3H]hemicholinum-3 (HC-3) and [3H]raclopride were used to label muscarinic cholinergic receptors, high-affinity choline uptake sites and dopamine D2 receptors, respectively. The degeneration of nigrostriatal pathway produced a transient decrease in [3H]QNB binding in the parietal cortex of both ipsilateral and contralateral sides at 2 and 8 weeks postlesion. [3H]QNB binding also showed a mild but insignificant decrease in the ipsilateral striatum throughout the postlesion periods. No significant change was observed in the substantia nigra (SN) of both ipsilateral and contralateral sides throughout the postlesion periods. In contrast, [3H]HC-3 binding showed no significant change in the parietal cortex of both ipsilateral and contralateral sides during the postlesion. However, [3H]HC-3 binding was upregulated in the ipsilateral dorsolateral striatum throughout the postlesion periods. The ventromedial striatum also showed a significant increase in [3H]HC-3 binding at 1 week and 2 weeks postlesion. On the other hand, no significant change in [3H]raclopride binding was found in the parietal cortex of both ipsilateral and contralateral sides during the postlesion. [3H]Raclopride binding showed a conspicuous increase in the ipsilateral striatum (35-52% of the sham-operated values in the lateral part and 39-54% in the medial part) throughout the postlesion periods. In the contralateral side, a mild increase in [3H]raclopride binding was also found in the striatum (10-15% of the sham-operated values in the lateral part and 22% in the medial part) after lesioning. However, a significant decline in [3H]raclopride binding was observed in the ipsilateral SN and ventral tegmental area during the postlesion. The present study indicates that 6-hydroxydopamine injection of medial forebrain bundle in rats can cause functional changes in high affinity choline uptake site in the striatum, as compared with muscarinic cholinergic receptors. Furthermore, our studies demonstrate an upregulation in dopamine D2 receptors in the striatum and a decrease in the receptors in the SN and ventral tegmental area after the 6-hydroxydopamine injection. Thus, these findings provide further support for neurodegeneration of the nigrostriatal pathway that occurs in Parkinson's disease. PMID- 11041270 TI - Presynaptic modulation of sympathetic neurotransmission in rat left ventricle slices: effect of pressure overload. AB - The purpose of this study was to establish the rat left ventricle (LV) tissue slice system for examination of norepinephrine (NE) release from sympathetic nerve terminals. Moreover, initial experiments were performed to use the LV tissue slice system to examine differences in NE uptake and release following cardiac pressure overload induced by abdominal aortic constriction (AC). Kinetic parameters (Vmax, Km) for the specific uptake of [3H]-NE demonstrated high affinity (Km, 1.94 +/- 0.83 microM) and moderate capacity uptake (Vmax, 182 +/- 6 fmol/mg/weight/min). Following 10 days of pressure overload, the Vmax for [3H]-NE uptake was significantly reduced (by 46%) in LV slices from AC rats compared to sham-operated (SO) controls. In control rat LV slices preloaded with [3H]-NE, electrically evoked [3H]-overflow was calcium- and stimulus pulse number dependent. The neuronal uptake inhibitor, desipramine (DMI), increased (by 60%) evoked [3H]-overflow from LV slices. The alpha2-agonist, UK14304, decreased evoked [3H]-overflow from LV slices in a concentration-dependent manner (maximal reduction of 75%). The beta2-agonist, salbutamol, increased evoked [3H]-overflow from LV slices in a concentration-dependent manner (maximal increase of 200%). In separate experiments, the LV tissue slice system was used to examine the effect of pressure overload on evoked [3H]-overflow. Following 10 days of pressure overload, evoked [3H]-overflow from LV slices of AC rats was increased (by 50%) compared to SO control. Increases in evoked [3H]-overflow from LV slices of AC rats compared to SO controls remained evident in the presence of DMI. These results demonstrate the relative importance of NE release and uptake using an in vitro LV tissue slice system. Sympathetic nerve terminals innervating rat LV were demonstrated to possess functional presynaptic alpha2- and beta2-adrenergic receptors. Finally, using this LV tissue slice system, reductions in the uptake velocity and increases in evoked NE release were demonstrated in response to acute cardiac pressure overload. PMID- 11041271 TI - Non-serotonergic potentiation by (-)-pindolol of DOI-induced forward locomotion in rats: possible involvement of beta-adrenoceptors?. AB - [1] We have previously shown that the beta-adrenergic/5-HT1 receptor partial agonist (-)-pindolol (2.0-32.0 micromol kg(-1)) enhances the increase in forward locomotion in rats produced by the 5-HT2 receptor agonist DOI (0.7 micromol kg( 1)) via net activation of post-synaptic 5-HT2 receptors. [2] It was found that neither the 5-HT1A receptor agonist and partial agonist, (+/-) 8-OH-DPAT (0.2-2.4 micromol kg(-1)) and (S)-(-)-UH-301, respectively, nor the 5-HT1A receptor antagonist WAY-100635 (0.09-1.5 micromol kg(-1)), substituted for (-)-pindolol in this in vivo behavioral model. [3] This also applies to the 5-HT1B receptor agonist and antagonist anpirtoline (0.3-4.0 micromol kg(-1)) and isamoltane (1.0 64.0 micromol kg(-1)), respectively. Neither of these compounds mimicked (-) pindolol in its interactions with DOI. [4] The (-)-pindolol/DOI-induced increase in forward locomotion could be antagonized by the beta1 adrenoceptor antagonist betaxolol (24 micromol kg(-1)). [5] It is suggested that the intrinsic efficacy of (-)-pindolol at beta-adrenoceptors is an important aspect of its in vivo pharmacodynamic profile. PMID- 11041272 TI - Effect of heat stress on serotonin-2A receptor-mediated intracellular calcium mobilization in rat C6 glioma cells. AB - This study was conducted to investigate an effect of heat stress at 44 degrees C for 30 min on intracellular Ca2+ signaling system and on heat shock protein (HSP) 70 expression. 5-HT-induced Ca2+ mobilization was reduced 1, 3 and 6 hrs after heat stress, and recovered to the control level 12 and 24 hrs after heat stress. One hr after heat stress, Ca2+ rise was significantly decreased when the cells were stimulated by any concentration of 5-HT. Thrombin-induced Ca2+ increase was also markedly reduced 1 hr after heat stress. HSP-70 level was increased 6 and 9 hr after heat stress. In HSP synthesis inhibitor quercetin-treated cells, HSP-70 expression was not enhanced after heat stress, and Ca2+ rise in response to 5-HT did not return to the control level. However, the Ca2+ rise induced by 5-HT was not restored to the control level after stress in Ac-Asp-Glu-Val-Asp-H (DEVD) exposed cells while DEVD had little effect on heat stress-induced synthesis of HSP-70. Dexamethasone did not alter the change in HSP-70 expression or Ca2+ response after heat stress. These results indicate that heat stress attenuated 5 HT-induced Ca2+ mobilization and that HSP-70 expression played an important role in recovery from Ca2+ impairment, possibly via protease activity in C6 cells. PMID- 11041273 TI - Involvement of other neurotransmitters in behaviors induced by the cannabinoid CB1 receptor antagonist SR 141716A in naive mice. AB - The receptor mechanisms by which the selective cannabinoid CB1 receptor antagonist/inverse agonist, SR 141716A [N-piperidino-5-(4-chlorophenyl)-1-(2,4 dichlorophenyl)-4-methyl-3-pyraz ole-carboxamide] produces scratching and head twitch response (HTR) in naive mice were examined. Acute intraperitoneal administration of varying doses of SR 141716A produced both scratchings (ED50 = 3.9 mg/kg) and head-twitches (ED50 = 4.6 mg/kg) in a dose-dependent manner. A dose of 10 mg/kg SR 141716A was used to induce the cited behaviors for drug interaction studies. The selective 5-HT2A/C receptor antagonist, SR 46349B [trans 4-[(3Z)3-(2-dimethylaminoethyl) oxyimino-3-(2-fluorophenyl) propen-1-yl] phenol] potently and completely blocked the head-twitches produced by SR 141716A (ID50 = 0.08 mg/kg). The induced scratching behavior was partially (68%) and less potently (ID50 = 0.6 mg/kg) blocked by SR 46349B pretreatment. The AMPA/kainate receptor antagonist, CNQX [6-cyano-7-nitroquinoxaline-2,3-dione], partially attenuated (68-78%) the induced scratching and head-twitching behaviors. On the contrary, the selective NMDA antagonist, AP-3 [(+/-)-2-amino-3-phosphonopropionic acid], had no significant effect on these behaviors. The selective tachykinin NK1 antagonist, CP 94, 994 [(+/-)-(2S, 3S)-3-(2-methoxybenzylamino)-2 phenylpiperidine], also partially attenuated both the scratching (64%) and the head-twitching (76%) symptoms produced by SR 141716A. Since SR 141716A lacks affinity for the discussed receptors, it appears that the induction of the cited behaviors probably involve indirect activation of their respective neurotransmitter systems. PMID- 11041274 TI - The influence of MK-801 on bicuculline evoked seizures in adult mice exposed to transient episode of brain ischemia. AB - The aim of the study was to examine the role of NMDA receptors in modulation of protective effect against bicuculline toxicity after transient brain ischemia in mice. Animals were exposed for 30 min to bilateral clamping of the common carotid arteries (BCCA) in anaesthesia. MK-801 was administered intraperitoneally in two paradigms: a) acute treatment: twice, 1.0 mg/kg; 1 hour before clamping and 6 hours after re-circulation and b) chronic treatment: 0.1 mg/kg; started 24 hours after re-circulation and continued once daily for 13 days, the last injection was administered 24 hours before seizure induction. 14 days after BCCA, the animals were injected with bicuculline (3.5 mg/kg s.c). A significant decrease in seizure susceptibility could be observed in BCCA treated mice compared with sham-operated controls. Acute treatment with MK-801 did not affect seizure activity both in sham and BCCA mice. Chronic treatment with the drug potentiated anticonvulsant effect of brain ischemia but had no influence on seizure activity in sham operated mice. The analysis of GABA content in brain tissue performed 14 days after BCCA showed a moderate increase in vehicle-treated mice and significant elevation after chronic treatment with MK-801. It can be suggested that NMDA receptors are not involved in the induction of a protective effect against bicuculline toxicity after transient brain ischemia. The prolonged treatment with low doses of MK-801 may potentiate a developed process in a mechanism of chemical preconditioning. PMID- 11041275 TI - Oxidative metabolites of 5-S-cysteinyldopamine inhibit the alpha-ketoglutarate dehydrogenase complex: possible relevance to the pathogenesis of Parkinson's disease. AB - A characteristic change in the substantia nigra of Parkinson's disease patients is an apparent accelerated rate of dopamine oxidation as evidenced by an increased 5-S-cysteinyldopamine (5-S-CyS-DA) to dopamine ratio. However, 5-S-CyS DA is more easily oxidized than dopamine to give 7-(2-aminoethyl)-3,4-dihydro-5 hydroxy-2H-1,4-benzothiazine-3-carboxylic acid (DHBT-1). Previous studies have demonstrated that DHBT-1 can be accumulated by intact rat brain mitochondria and inhibits complex I but not complex II respiration. In this study, it is shown that DHBT-1 also inhibits the alpha-ketoglutarate dehydrogenase complex (alpha KGDH) but not cytochrome c oxidase (complex IV). The inhibition of alpha-KGDH is dependent on the oxidation of DHBT-1, catalyzed by an unknown constituent of the inner mitochondrial membrane, to an electrophilic o-quinone imine that covalently modifies active site sulfhydryl residues. The latter conclusion is based on the ability of > or = equimolar glutathione to block the inhibition of alpha-KGDH by DHBT-1, without altering its rate of mitochondrial membrane-catalyzed oxidation, by scavenging the electrophilic o-quinone intermediate forming glutathionyl conjugates which have been isolated and spectroscopically characterized. Activities of mitochondrial alpha-KGDH and complex I, but not other respiratory complexes, are decreased in the parkinsonian substantia nigra. Such changes together with evidence for accelerated dopamine oxidation, increased formation of 5-S-CyS-DA and the ease of oxidation of this conjugate to DHBT-1 which inhibits alpha-KGDH and complex I, without affecting other respiratory enzyme complexes, suggests that the latter putative metabolite might be an endotoxin that contributes to the alpha-KGDH and complex I defects in Parkinson's disease. PMID- 11041276 TI - Paraoxonase polymorphisms, pesticide exposure and Parkinson's disease in a Caucasian population. AB - Parkinson's disease (PD) has been associated with exposure to pesticides and oxidative injury. The involvement of paraoxonase in both pesticide metabolism and lipid peroxidation suggests that it may play a role in the pathogenesis of PD. We examined the frequency of polymorphic alleles of the PON1 and PON2 genes in a sample of caucasian subjects with PD. The frequency distribution of these genotypes did not differ significantly between patients and controls, including those who had reported exposure to pesticides. PMID- 11041277 TI - N-methylation ability for azaheterocyclic amines is higher in Parkinson's disease: nicotinamide loading test. AB - The discovery of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) leads to the hypothesis that Parkinson's disease (PD) is may be initiated or precipitated by endogenous toxins by the mechanism similar to that of MPTP in genetically predisposed individuals. The higher cerebrospinal fluid levels of N-methylated azaheterocyclic amines, such as beta-carboline and tetrahydroisoquinoline, have been found in parkinsonian patients compared with age-matched controls. To estimate the N-methylation ability for azaheterocyclic amines in parkinsonian patient, nicotinamide was dosed with 100 mg to 26 parkinsonians and 20 controls consisted of 16 other neurogenic disease patients and 4 healthy volunteers. The urine was collected for 4 h, and then analyzed urinary its metabolites by an improved HPLC method. Nicotinamide has a pyridine ring in its structure and may be metabolized through the pathways similar to those for the endogenous neurotoxins. The urinary excretions of nicotinamide metabolites were significantly affected by aging. The excretion of N1-methylnicotinamide decreased along with aging both in PD patients and controls. In younger (65 years old or younger) PD patients, the excretion amount of N1-methylnicotinamide was significantly higher than that in younger controls. The decline rate of N1 methylnicotinamide excretion in parkinsonians was significantly greater than that in controls; the rate is more than 2-fold higher in parkinsonian patients. The age-associated decrease in 1-methyl-2-pyridone-5-carboxyamide excretion was observed only in parkinsonian patients, but not in controls. The total excreted amount of N-methylated metabolites (N1-methylnicotinamide plus 1-methyl-2 pyridone-5-carboxyamide) was also observed the age-related decline in both groups. The urinary excretions of nicotinamide and nicotinamide-N-oxide were not influenced by aging. These results would indicate that the excess N-methylation ability for azaheterocyclic amines before the onset had been implicated in PD. On the other hand, the present results suggested that the contribution of aberrant cytochrome P450 or aldehyde oxidase activity acting on the pyridine ring, that could act as detoxification routes of endogenous neurotoxins, would be small in the etiology of PD. PMID- 11041278 TI - ECT in Parkinson's disease-dopamine transporter visualised by [123I]-beta-CIT SPECT. AB - Parkinson's disease (PD) is characterised by a loss of dopaminergic neurones in the basal ganglia. These neurones may be visualised by single photon emission computed tomography (SPECT) with the cocaine analogue 2beta-carboxymethyl-3-beta (4-iodophenyl)tropane ([123I]beta-CIT), which labels the dopamine reuptake sites in the nerve terminals. In order to evaluate the possibility to predict the outcome of ECT a prospective study was performed with six PD patients in whom the [123I]beta-CIT uptake was measured before and after an electroconvulsive therapy (ECT) series. The side-to-side difference in the radiotracer uptake was found to be significantly lower in striatum located contralaterally to the part of the body with the most pronounced symptomatology. No significant change in uptake of the radioligand was seen after ECT. Patients with best uptake and thus with less advanced PD improved most after ECT. The possibility to use the [123I]beta-CIT uptake to predict the outcome of ECT treatment has to be further evaluated. PMID- 11041279 TI - Antidopaminergic effects of 1,2,3,4-tetrahydroisoquinoline and salsolinol. AB - Immediate behavioral and biochemical effects of single doses of 1,2,3,4 tetrahydroisoquinoline (TIQ, 50 mg/kg) and salsolinol (100 mg/kg), suspected of involvement in etiology of Parkinson's disease, were investigated. Apomorphine (0.25 mg/kg) or haloperidol (1 mg/kg) were administered to TIQ or salsolinol pretreated Wistar rats. In additional experiment the displacement of [3H]apomorphine by TIQ, salsolinol and dopamine receptor agonists and antagonists was tested. Both tetrahydroisoquinolines only slightly affected behavior and dopamine metabolism in naive rats, but very effectively abolished the behavioral and biochemical effects of apomorphine (hyperactivity, depression of striatal HVA level). The behavioral and biochemical effects of haloperidol were unchanged by administration of TIQ nor salsolinol. The tetrahydroisoquinolines displaced [3H]apomorphine from its binding sites with effectiveness comparable to that of dopamine. The results support the hypothesis that endogenous tetrahydroisoquinolines may play an important role in regulation of dopaminergic activity in non-senescent organisms. PMID- 11041280 TI - Serum levels of coenzyme Q10 in patients with amyotrophic lateral sclerosis. AB - To elucidate whether serum coenzyme Q10 levels are related with the risk for amyotrophic lateral sclerosis (ALS), we compared serum levels of coenzyme Q10 and the coenzyme Q10/cholesterol ratio, in 30 patients with ALS and 42 matched controls using a high performance liquid chromatography technique. The mean serum coenzyme Q10 levels and the coenzyme Q10/cholesterol ratio did not differ significantly between the 2 study groups. These values were not influenced by the clinical form (spinal vs. bulbar) of ALS, and they did not correlate with age, age at onset, and duration of the disease. These results suggest that serum coenzyme Q10 concentrations are unrelated with the risk for ALS. PMID- 11041281 TI - Membrane breakdown in acute and chronic neurodegeneration: focus on choline containing phospholipids. AB - Breakdown of cellular membranes is a characteristic feature of neuronal degeneration in acute (stroke) and chronic (senile dementia) neurological disorders. The present review summarizes recent experimental and clinical work which concentrated on changes of choline-containing phospholipids as indicators of neuronal membrane breakdown. Experimental studies identified glutamate release, calcium influx, and activation of cellular phospholipase A2 (PLA2) as important steps initiating membrane breakdown in cultured neurons or brain slices under hypoxic or ischemic conditions. Proton NMR studies have shown an elevation of choline-containing compounds in the brain of Alzheimer patients while neurochemical studies in post mortem-brain demonstrated increases of the catabolic metabolite, glycerophosphocholine, an indicator of PLA2 activation. In contrast, studies of cerebrospinal fluid, phosphorus NMR studies, and measurements of phospholipases in post mortem Alzheimer brain gave ambiguous results which may be explained by methodical limitations. The finding that, in experimental studies, choline was a rate-limiting factor for phospholipid biosynthesis has stimulated clinical studies aimed at counteracting phospholipid breakdown, e.g. by combinations of choline and cytidine. Future experimental approaches should clarify whether loss of membrane phospholipids is cause or consequence of the neurodegenerative disease process. PMID- 11041282 TI - No association between the alpha2-macroglobulin (A2M) deletion and Alzheimer's disease, and no change in A2M mRNA, protein, or protein expression. AB - A polymorphism consisting of a deletion near the 5' splice site of exon 18 on the alpha2-macroglobulin (A2M) gene (A2M-2) has been suggested to be associated with Alzheimer's disease (AD) in family-based studies. We studied the A2M-2 allele together with the ApoE alleles in a large series on patients with AD (n = 449) and age-matched controls (n = 349). Neuropathologically confirmed diagnoses were available in 199 cases (94 AD and 107 control cases). We found no increase in A2M 2 genotype or allele frequencies in AD (27.5% and 14.6%) versus controls (26.4% and 14.9%). In contrast, a marked increase (p < 0.0001) in ApoE epsilon4 genotype or allele frequencies was found in AD (66.6% and 41.2%) as compared with controls (29.8% and 16.5%), suggesting sufficient statistical power in our sample. No relation was found between the A2M-2 and the ApoE epsilon4 allele. No change in A2M exon 17-18 mRNA size or sequence or A2M protein size was found in cases carrying the A2M-2 deletion, suggesting that there is no biological consequences of the A2M intronic deletion. No change in A2M protein level in cerebrospinal fluid was found in AD, suggesting that the A2M-2 allele does not effect the A2M protein expression in the brain. The lack of an association between the A2M-2 allele and AD in the present study, and the lack of abnormalities in the A2M mRNA or protein suggest that the A2M-2 allele is not associated with AD. PMID- 11041283 TI - No evidence for an association between the Glu298Asp polymorphism of the NOS3 gene and Alzheimer's disease. AB - Recently a significant association of a missense mutation (Glu298Asp) of the endothelial nitric oxide synthase (NOS3) gene with late-onset Alzheimer's disease (LOAD) was reported. We tried to replicate this finding in a Japanese sample of 121 patients with LOAD, 51 with early-onset AD (EOAD), and 165 medical controls. However, the genotype and allelic distributions for the Glu298Asp polymorphism were similar for these three groups, suggesting that the Glu298Asp polymorphism of the NOS3 gene has no relevance to the development of AD in Japanese. PMID- 11041284 TI - Reduction of the synaptic protein rab3a in the thalamus and connecting brain regions in post-mortem schizophrenic brains. AB - Although the psychotic symptoms in schizophrenia can be alleviated by treatment with dopaminergic receptor antagonists, the etiology and underlying neurochemical pathology remains obscure. Both neuropathological and magnetic resonance imaging studies have found evidence for neuronal loss and atrophy in the thalamus in schizophrenia, implicating this key structure for gating information to cortical areas in the pathophysiology. Recent studies have also found evidence of synaptic loss in the thalamus in schizophrenia. To further examine possible synaptic disturbances, we studied the synaptic related protein rab3a as a marker for synaptic density, using both quantitative Western blotting and immunohistochemistry. The material consisted of brains from 22 schizophrenic patients (mean age 79.3 years), and 24 control subjects (74.8 years). Reduced rab3a protein levels were found in the left thalamus in schizophrenia (0.47 +/- 0.17 vs. 1.00 +/- 0.18; p < 0.0001), while a less marked decrease was found also in the right thalamus (0.75 +/- 0.13 vs. 1.00 +/- 0.09; p < 0.0001). Immunohistochemistry, performed on two schizophrenic and two control brains, revealed that rab3a immunoreactivity was most reduced in the left anterior and mediodorsal thalamic nuclei. Therefore, we extended the study to brain regions connected these thalamic nuclei. Reduced rab3a protein levels were found schizophrenia also in the frontal cortex, hippocampus, gyrus cinguli, and parietal cortex, while no significant differences were found in the temporal cortex, or in cerebellum. The reduction in rab3a was not found to be secondary to confounding factors such as age-differences, post-mortem delay time, generalized brain atrophy, or antipsychotic medication. Therefore, the reduction of rab3a probably reflects synaptic disturbances, possibly synaptic loss, in the limbic system and neocortical areas, in schizophrenia. This part of the brain is known to be involved in behavioral and emotional control, and thus to be crucial for higher mental functions, suggesting that synaptic disturbances in the limbic system may be of importance in the development of psychotic symptoms in schizophrenia. PMID- 11041285 TI - Co-administration of furosemide augments tacrolimus-induced impairment in kidney function in rats. AB - Sodium-depletion in rats reproduces functional and morphological tacrolimus nephrotoxicity observed in man. Potent diuretics induce sodium-depletion. Our objective was to determine the effect of a loop diuretic furosemide on tacrolimus mediated functional and pathological impairment of the kidney in rats. Sprague Dawley rats were divided into four groups; group 1, rats received vehicle (saline) only; group 2, rats were treated with tacrolimus (1 mg/kg body weight) and furosemide (5 mg/kg body weight); group 3, rats were treated with tacrolimus alone; and group 4, rats were treated with furosemide (5 mg/kg body weight) alone. On day 28, tail blood pressure was measured and the rats were placed in metabolic cages for urine collection. After 24 hr the rats were sacrificed. Tacrolimus alone tended to cause growth retardation, hypotension, hypomagnesemia and a rise in blood urea nitrogen. Furosemide co-administration enhanced the effects of tacrolimus on hypotension, hypomagnesemia and a rise in blood urea nitrogen. The renal histology characterized by cytoplasmic vacuolization of the proximal tubules was not different between the rats treated with both tacrolimus and furosemide and the rats treated with tacrolimus alone. A strong immunostaining for FKBP-12, a tacrolimus-binding protein, was observed in the medulla of the kidneys of rats treated with tacrolimus either with or without furosemide. These results indicate that furosemide further augments tacrolimus induced impairment in kidney function, and that furosemide should be used with discretion in patients on tacrolimus therapy. PMID- 11041286 TI - Does captopril change oxidative stress in puromycin aminonucleoside nephropathy? AB - Puromycin aminonucleoside (PAN) nephropathy in rats has been induced by the intraperitoneal injections of PAN. One group of animals which received PAN has been treated simultaneously with captopril (angiotensine converting enzyme-ACE inhibitor) with the aim to test whether continuing treatment with captopril along with PAN injections would be able to modulate the toxic effects of PAN. The third group of rats was given only captopril. Morphological changes in the kidney were evaluated by scanning electron microscopy that showed the loss of podocyte foot processes in the kidney of PAN treated animals but also in the kidney of captopril treated ones as well as in the animals treated with both drugs simultaneously. Reduced glutathione content, catalase, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), xanthine oxidase activities as well as lipid peroxides were investigated in rat blood and kidney. Captopril given alone produced a significant decrease of plasma lipid peroxides, but it did not show any significant effect on investigated antioxidative factor levels neither in blood nor in the kidney. PAN given alone produced a significant depletion of plasma lipid peroxides, kidney catalase and erythrocyte GSH-Px activity as well as a significant increase of plasma catalase and erythrocyte SOD activity. Treatment of animals with both drugs simultaneously resulted in a significant increase of erythrocyte SOD activity and a significant decrease of plasma lipid peroxides, erythrocyte GSH-Px and kidney SOD activities. Kidney xanthine oxidase activity showed a significant increase in both PAN and PAN plus captopril treated animals in comparison with the values of captopril treated rats. These data suggest that PAN changes the antioxidative factor pattern in rat blood and kidney. Contrary to our expectations that captopril may protect the toxic effects of PAN it only to a certain extent modifies these effects showing protective effect only on tissue catalase activity. PMID- 11041287 TI - Effect of lithium on cyclosporin induced nephrotoxicity in rats. AB - Psychoactive drugs provide essential intervention in the care of transplant recipients, yet little is known of their interaction with immunosuppressants such as cyclosporin (CSA). Lithium (Li) is an invaluable drug for the treatment of manic disorders in organ transplant patients. As both these drugs are known to produce renal toxicity, the concomitant use of CSA and Li may be potentially harmful. The present study was undertaken to investigate the effect of CSA and Li chloride individually and in combination on renal structure and function of rats. Male Sprague-Dawley rats were divided into the following eight groups of seven animals each: group I, control (vehicle only); group 2, Li (2 mEq/ kg i.p.) alone; group 3, CSA 12.5 mg/kg (subcutaneous); group 4, CSA 25 mg/kg; group 5, CSA 50 mg/kg; group 6, CSA 12.5 mg/kg + Li; group 7, CSA 25 mg/kg + Li; and group 8, CSA 50 mg/kg + Li. The drugs were given once a day for seven days; Li being administered 30 min before CSA. Twenty four hours after the last dose of drugs the animals were sacrificed and blood samples were analyzed for blood urea nitrogen (BUN), serum creatinine (SCr), CSA and Li levels. The left kidney was analyzed for malondialdehyde (MDA) and conjugated dienes (CD) levels and right kidney was used for histopathological studies. Our results showed that Li alone did not produce any significant renal toxicity, whereas CSA dose dependently caused structural and functional changes in kidneys. However, significantly higher structural and functional impairment was observed in the animals treated with Li plus CSA as compared to CSA alone treated animals. Several fold increase in blood Li level was also noticed in the rats concomitantly treated with CSA and Li. A significant increase in MDA and CD in the rats treated with CSA plus Li suggests the role of oxidative stress in drug induced nephrotoxicity. These findings clearly demonstrate that even non toxic doses of Li may significantly exacerbate CSA induced nephrotoxicity in rats. The enhanced nephrotoxicity following concomitant use of these drugs may be attributed to significant increase in the bioavailability of Li and enhanced oxidative stress. Further clinical studies are warranted to investigate the interaction of these nephrotoxic drugs in human subjects. PMID- 11041288 TI - Effects of hypokalemia on cardiac growth. AB - In neonatal myocytes grown in culture, reductions in extracellular potassium concentration produced a hypertrophic response as assessed by induction of early response genes, atrial natriuretic peptide and skeletal actin, and repression of the alpha3 isoform of the sodium pump in a dose dependent manner. The degree of alpha3 repression appeared to be dose dependent with decreases in media (K). Similarly, decreases in media potassium concentrations caused increases in cytosolic calcium concentration in a dose dependent manner; moreover these increases in cytosolic calcium concentration correlated quite well with repression of alpha3 expression. In contrast, although moderate reductions of potassium concentration induced upregulation of skACT and ANP, severely reduced potassium concentrations caused repression of skACT and ANP expression. In parallel studies performed in vivo, 3-5 weeks dietary K restriction induced molecular phenotypical changes similar to that seen in the neonatal myocyte model without demonstrable growth as assessed by the heart weight/body weight ratio. However, when rates subjected to dietary K restriction were subsequently subjected to acute aortic constriction, cardiac growth was greater than in rats fed a control diet. These data suggest that hypokalemia may produce molecular phenotypic alterations consistent with cardiac hypertrophy as well as contribute to hypertrophy in an in vivo model. PMID- 11041289 TI - Perinatal complications and three-year follow up of infants of diabetic mothers with diabetic nephropathy stage IV. AB - The objective of the study was to evaluate differences in the perinatal complications and in the 3-year follow up of infants of diabetic mothers with and without diabetic nephropathy stage IV. We compared the fetal and maternal complications and the early postpartal development until 3 years after delivery in 10 children of nephropathic diabetic mothers and 30 children of diabetic mothers without nephropathy. The mean (+/-SD) birthweight of the infants of nephropathic women was 2,250 +/- 496 g versus 3,544 +/- 435 g in the women without nephoropathy (p < 0.01). Births were premature in six pregnancies (60%) of the nephrotic women but in none of the women without nephropathy (p < 0.01). Three infants (30%) of the women with nephropathy showed respiratory distress syndrome in contrast to two babies (6%) of the women without nephropathy. Pre eclampsia or eclampsia occurred in 6 (60%) pregnant women with and in two women (6%) without diabetic nephropathy (p < 0.01). Nephrotic syndrome was observed in 7 nephrotic women (70%) in contrast to none women without nephropathy. Three years postpartum, six of the children (60%) of nephropathic women had a body weight < the 50th percentile but none of the children of the women without nephropathy did so (p < 0.01). In addition, the children of nephropathic mothers started to speak significantly later (15 +/- 3 versus 12 +/- 13 months postpartum, p < 0.05) and had infectious diseases more commonly (60% versus 6%, p < 0.01) than the children of women without nephropathy. It can be concluded that in pregnancies of diabetic women the birth weights of the infants are significantly smaller and the fetal as well as maternal complication-rates significantly higher than in those of women without nephropathy. Also 3 years after delivery, the body weight of the children of nephropathic diabetic women is significantly lower than that of children of diabetic women without nephropathy. Additionally, children of nephropathic women are retarded in terms of linguistic development and their resistance to infections is reduced. PMID- 11041290 TI - The predictive value of 131I-hippurate clearance in the prognosis of acute renal failure. AB - The aim of this investigation was to study the validity of the radionuclide methods in the estimation of kidney function, for prognosis and follow-up of acute renal failure (ARF). In thirty-one ARF patients, the evaluation of glomerular filtration rate (GFR) by 99mTc-DTPA clearance and effective renal plasma flow (ERPF) by 131 I-ortoiodohippurate (131I-OIH) clearance was performed within 7 days and after 6 months from ARF onset. All patients were divided in three groups according to 131I-OIH clearance values obtained within 7 days: group 1, under 150 mL/min; group 2, 150-250 mL/min; and group 3, over 250 mL/min. Seven days clearance values of both radiopharmaceuticals were found to be very low, however, GFR was found more severely impaired than ERPF. Clearance values obtained after 6 months demonstrated no recovery of renal function in the first group, partial recovery in the second and almost complete recovery in the third group. Patients with the lowest 131I-OIH clearance values at the ARF onset had no recovery of renal function, while in the other two groups recovery corresponded to initial 131I-OIH clearance values. In patients with ARF both, 99mTc-DTPA and 131I-OIH clearances were shown suitable for the follow up of renal function, however, only 131I-OIH clearance had a strong predictive prognostic value for renal function recovery in ARF. PMID- 11041291 TI - The heterogeneous and delayed course of blood pressure normalization in hypertensive patients after bilateral nephrectomy with and without subsequent renal transplantation. AB - BACKGROUND: Controversy exists about the time course of blood pressure normalization following bilateral nephrectomy. We sought to evaluate the time course of blood pressure normalization following bilateral nephrectomy and after subsequent kidney transplantation. METHODS AND RESULTS: Clinical data from 14 hypertensive patients were retrospectively assessed. Baseline blood pressure was 175 +/- 33/109 +/- 9 mmHg. Ten patients firstly underwent unilateral nephrectomy, which resulted in a slight increase of blood pressure (185 +/- 22/110 +/- 5 mmHg). One month following bilateral nephrectomy, blood pressure was 167 +/- 23/104 +/- 17 mmHg, at 3 months 159 +/- 42/104 +/- 25 mmHg, and at 6 months 149 +/- 41/96 +/- 30 mmHg. Antihypertensive medication was necessary in 9/14 patients at a 2 year follow-up. Eight patients remained anephric (group I), 6 patients had subsequent kidney transplantation (group II). In group I, blood pressure was 159 +/- 42/93 +/- 17 mmHg and 129 +/- 34/75 +/- 14 mmHg at 3 and 6 months, respectively (p< 0.05 vs. baseline). In group II, blood pressure decreased from 188 +/- 42/ 128 +/- 46 mmHg to 167 +/- 48/113 +/- 32 mmHg at 3 months, but increased after transplantation to 186 +/- 39/118 +/- 33 mmHg. Antihypertensive medication was still necessary in 5 transplanted patients (83%) and in 3 anephric patients (38%). CONCLUSION: Adaptation of the blood pressure response following bilateral nephrectomy is a time requiring process, and long-term antihypertensive medication may still be necessary. PMID- 11041292 TI - Outcome of dialysis patients submitted to coronary revascularization. AB - Cardiovascular disease accounts for almost half of the total mortality in patients with end stage renal disease (ESRD). It has recently been debated whether coronary revascularization has the same rate of risks and successes in this cohort of patients compared to patients without renal disease. Since 1991, 17 dialysis patients were submitted to coronary revascularization in our center. Seven patients were following peritoneal, 10 hemodialytic treatment. Four patients were submitted to percutaneous transluminal coronary angioplasty (PTCA) and 13 to surgical revascularization (CABG). In 2 patients the coronary lesion was unique, in the others stenosis of multiple vessels were found. Six patients were diabetic. The mean age at the onset of the coronary artery disease (CAD) was 57.17 +/- 11.6 years. The mean time elapsed from the onset of the CAD and the performance of the PTCA or CABG was 30.1 +/- 35.4 months. The mean time from beginning of dialysis treatment to revascularization was 48.2 +/- 39.6 months. Mean hemoglobin values were 9.7 +/- 1 g/dL, mean phosphorus values were 5.2 +/- 8.7 mg/dL, mean cholesterol values were 211 +/- 49.5 mg/dL. The procedure was technically successful in all patients. Mean survival was 25.09 +/- 28.12 months. Twelve patients died, 5 of whom within one month. Survival at one month was 70.5%, at 6 months 58.8%, at one year 52.9%, at 2 years 47%. There was neither significant difference patients submitted to PTCA and those submitted to CABG, nor between diabetic and non-diabetic patients. In conclusion, coronary revascularization in our experience is a high risk procedure in dialysis patients. The reasons for this could be the severe general conditions of these patients affected with diffuse vasculopathy and the long time elapsed since the onset of the ischemic cardiopathy. Thus, our results could suggest the opportunity of performing earlier screening of coronary situation and revascularization treatment in CAD dialysis patients. PMID- 11041294 TI - Acute renal failure associated with liver disease in India: etiology and outcome. AB - Acute renal failure (ARF) associated with liver disease is a commonly encountered clinical problem of varied etiology and high mortality. We have prospectively analyzed patients with liver disease and ARF to determine the etiology, clinical spectrum, prognosis and factors affecting the outcome. Other than hepatorenal syndrome patients, out of 221 cases, 66 developed ARF secondary to various liver disease like cirrhosis (n = 29, mortality 8, risk factors-older age p < 0.01, grade III/IV encephalopathy p < 0.05), fulminant hepatic failure (n = 25, mortality 15, risk factor-prolonged prothrombin time p < 0.01), and obstructive jaundice (n = 12, mortality 7, risk factor-sepsis p < 0.01). In these three groups the factors leading to ARF were volume depletion (24), gastrointestinal bleed (28), sepsis (34), drugs (27) [aminoglycosides (9) and NSAID (18)] along with hyperbilirubinemia. Various types of ARF with contemporaneous liver injury were malaria (n = 37, mortality 15, risk factors-higher bilirubin p < 0.001, higher creatinine p < 0.05, anuria p < 0.05 and dialysis dependency p < 0.05), sepsis (n = 36, mortality 22, risk factors-age p < 0.001, higher bilirubin p < 0.01, oliguria p < 0.05), hypovolemia with ischemic hepatic injury (n = 14, mortality 5, risk factors-higher creatinine p < 0.05 and SGPT p < 0.01), acute pancreatitis (n = 12, mortality 4, risk factors-higher bilirubin p < 0.001, higher SGPT p < 0.01, dialysis dependency p < 0.05), rifampicin toxicity (n = 10, no mortality), paroxysmal nocturnal hemoglobinuria (n = 3, no mortality), CuSO4 poisoning (n = 3 mortality 2), post abortal (n = 11, mortality 6, risk factors higher creatinine p < 0.05 and SGPT p < 0.01), ARF following delivery including HELLP syndrome (n = 12, mortality 4, risk factors-higher bilirubin p < 0.01 and SGPT p < 0.01), and of uncertain etiology (n= 14 mortality 4). 133 patients (60.2%), required hemodialysis hemodialfiltration or peritoneal dialysis. ARF associated with liver disease is having high mortality (42.5%). Avoidance of dehydration, hypotension, nephrotoxic drugs and sepsis, with promote dialytic support are necessary to reduce mortality and morbidity. PMID- 11041293 TI - High frequency of amyloid lymphadenopathy in uremic patients. AB - Amyloid lymphadenopathy has only been reported in case report form, or in small groups of patient groups within large series. We believe that amyloid lymphadenopathy is common in uremic patients, and thus designed this study to determine the frequency of this condition in hemodialysis patients, and to assess its types and patterns. We reevaluated 46 uremic patients' lymph node biopsies for amyloid deposits. We also immunohistochemically identified the protein origin of these deposits using Amyloid A, kappa, lambda, beta2 microglobulin, and transthyretin antibodies. Histopathologically, we observed for vascular involvement, follicular deposition, and diffuse deposition. We detected amyloid deposits in 10 of the 46 (22%) patients' lymph nodes. The patterns of deposition were vascular involvement alone in six specimens, vascular involvement plus follicular deposition in three, and vascular involvement plus diffuse deposition in one specimen. Amyloid AA type protein was present in seven nodes, beta2 microglobulin-related amyloid in two nodes, and immunoglobulin-derived protein (AL) in one node. We assessed these 10 patients for causes of end-stage renal disease (ESRD) and other conditions that might relate to amyloidosis. The cause of ESRD in the seven patients with AA amyloid were renal amyloidosis secondary to Familial Mediterranean Fever in four, glomerulonephritis in one patient who had bronchiectasis and Castleman's disease, unknown in one patient who had bronchial asthma, and pyelonephritis in one patient who had no characteristics that could be linked with AA type amyloidosis. The causes of ESRD in the two individuals with beta2 microglobulin-related amyloidosis who had been on long-term hemodialysis were pyelonephritis and glomerulonephritis. The cause of ESRD in the patient with AL type protein was glomerulonephritis, and this patient had no systemic disease. We conclude that amyloid lymphadenopathy is, indeed, common in uremic patients. Amyloid type AA is the most prevalent form of amyloid protein in uremic patients, but amyloid type does not always correspond with underlying cause of renal failure, or with the presence of systemic disease. PMID- 11041295 TI - Acute renal failure complicating nonfulminant hepatitis a in HLA-B27 positive patient. AB - Here we report the case of a previously healthy 32-year-old HLA B27 positive male who developed completely reversible acute oliguric renal failure in the course of nonfulminant hepatitis A infection. This is a rare complication of uncertain etiology. B27 positive individuals are prone to a number of immune system derangements including overreacting in interactions with some microbiologic agents. We made our consideration on the basis of some experimental models and the presumed pathogenesis of reactive arthritis in these individuals. As a result here we postulate a hepatitis A virus-triggered, immune mediated mechanism of renal injury restricted to genetically susceptible (i.e. B27 positive) individuals. In regard to this hypothesis we warrant additional HLA-profiles and future similar cases for further confirms and conclusions. PMID- 11041296 TI - Unusual presentation of mesangial proliferative glomerulonephritis in HELLP syndrome associated with acute renal failure. AB - Acute renal failure in pregnancy is not common in industrialized countries. HELLP syndrome (hemolysis, elevated liver enzyme, and low platelets) was one of the causes of acute renal failure in pregnancy, but renal pathological findings in case of acute renal failure had rarely been reported. We reported an unusual case of HELLP syndrome with acute renal failure requiring renal replacement therapy and which histopathologic findings of kidney biopsy showed mesangial proliferative glomerulonephritis and her renal function completely recovered after immediate artificial abortion, supportive management, transfusion of blood products, and hemodialysis. PMID- 11041297 TI - Systemic lupus erythematosus presenting initially as hydrogen ATPase pump defects of distal renal tubular acidosis. AB - Tubulointerstitial involvement is well recognized in systemic lupus erythematosus. The tubular dysfunction is usually latent and usually presents after diagnosis of systemic lupus erythematosus. We report a case presenting that she is well previously and initially diagnosed as periodic paralysis of hypokalemia at emergency room and final diagnosis is systemic lupus erythematosus with H+-ATPase pump defect of distal type renal tubular acidosis. Kidney biopsy showed lupus nephritis classified as mesangial proliferative glomerulonephritis WHO class II B. Her renal tubular acidosis was subsided after steroid therapy was administered. PMID- 11041298 TI - Effect of polymyxin B immobilized fiber on encephalopathy in hemodialysis patients with sepsis. PMID- 11041299 TI - Quality dimensions for school psychology services. AB - Varying standards and quality among Norwegian school psychology services (SPS) have resulted in national SPS support and developmental programs. The present study aimed to reveal SPS quality dimensions to suggest a basis for development of service standards. Teachers (333) and administrators (136) responsible for corresponding cases referred to the SPS responded to a questionnaire including indices of case service quality and collaboration between schools and SPS. The statistical analyses included factor analyses and a multiple regression analysis. The data and results from an earlier study of parents' quality dimensions were included in an items analyses. The result suggests five central SPS quality dimensions: availability, participation, consideration, effectiveness and security. While effectiveness has the strongest effect on the evaluation of case service quality both for parents and school personnel, the dimensions have different significance for different consumer groups. Relations to general service quality dimensions and consequences of the findings are discussed. PMID- 11041300 TI - Assessment of weight and eating concerns in Norwegian adolescents. AB - The aim of the present study was to test the properties of an instrument that assesses concerns about weight and eating without reference to dieting behavior. A short instrument, the Weight and Eating Concerns Inventory (WECI) was examined in a sample of 569 boys and 548 girls aged 11 to 15 years. Confirmatory factor analyses with LISREL showed a better fit with a version of the instrument that did not include a reference to dieting behavior, compared to a version that included such a reference. This was true for both boys and girls, irrespective of their age group. However, the results indicated that both versions should be used with caution for young boys. The internal consistency of the WECI (that is, the version without reference to dieting) was satisfactory, ranging from 0.78 to 0.86 for girls and from 0.68 to 0.73 for boys. The WECI correlated quite substantially with negative self-evaluations and depression for boys and girls in all the age groups assessed, suggesting that high scores on the WECI may indicate a problem that goes beyond worries or concerns, and should be taken seriously. In general, girls reported more of these concerns than boys, and the correlation between the WECI and dieting was stronger among girls compared to boys. PMID- 11041301 TI - Chronic pain as a posture towards the world. AB - This article discusses what chronic pain is "about", what the intentional object is of pain, and what is the intentional relation like? My approach is based on Maurice Merleau-Ponty's phenomenology, with an aim is to understand a two-way relationship: how the sufferers bestow meaning on chronic pain, and how pain, on the other hand, signifies peoples' life. In contrast to biomedical and cognitive behavioral theories, chronic pain is not only meaningful, but as an intentional emotion as well; it does not simply "happen" in the nervous system. I analyzed meanings assigned to pain through the narratives of three patients with chronic pain. Pain is described as creating a discontinuity in the patient's Lebenswelt at the narrative level. When attempting to find meaning to their pain, patients point both to everyday life and biomedical referents. The structure of bestowing meaning is, metaphorically, like a necklace with everyday world and biomedical interpretations strung like beads, one after the other. The intentional object of pain, on the contrary, is constituted of the patients' world in its wholeness. My results don't confirm Drew Leder's idea of disrupted intentionality, but underline directness as the basic relation of human experience also in case of pain and disease. Pain in itself is an e-movere, an intense passionate movement, an intentional relation with and a bodily posture taken towards the world. PMID- 11041302 TI - The effect of strategies and contexts on memory for movement patterns. AB - Two experiments on memory for movement patterns examined possible motor aspects of this memory type. None of the experiments found any evidence for motor effects. The first experiment showed lower memory with induced motor strategies than with any of the other induced strategies. The second experiment demonstrated stronger context effects under retrieval than under encoding conditions. The results are discussed in terms of functional and structural conceptions of episodic short-term memory of motor information. PMID- 11041303 TI - The hidden meanings of metaphors in family therapy. AB - This study investigates how two therapists' beliefs and practices influence the therapeutic process when they organize social interaction according to a metaphor of a royal family. The therapeutic process is described through the case of a boy called Pelle. He comes to therapy together with his family. It is shown how the therapists collaborate in the process of implementing the worldview of the predefined normative standard for family life. In the short term the therapists' use of the metaphor can be seen as an intervention to accomplish immediate change in a non-threatening way. In the long term the cost of using the metaphor was that the mother got a confirmation about herself as a less powerful parent and the child got an image of being a failure. This study points out that metaphors as therapeutic tools have to be analyzed critically before they are used or more specifically the therapists have to examine what kind of values and meanings are hidden in the metaphor and who will gain and loose if it is used as an intervention. PMID- 11041304 TI - On the relationship between locus of control, level of ability and gender. AB - In a sample of Norwegian 14- and 15-year-old students no significant relationship was found between total externality-internality score and level of ability. However, a significant relationship was found between ability and a subscale of locus of control related to degree of belief in the impact of school effort. The analyses of gender differences showed that girls had significantly higher total internal locus of control scores than boys. Boys were, however, significantly more internally oriented than girls on a subscale related to the respondents' general belief in luck, while girls were significantly more internally oriented than boys on a school effort scale. The present study does not support the notion that girls develop an attributional pattern which is more closely related to their abilities while boys may develop a broader attributional pattern. PMID- 11041305 TI - Dimensions of fatigue in different working populations. AB - Perceived fatigue related to work has often been measured in one dimension. The main purpose of the present study was to validate a proposed five-factor model of perceived fatigue in a new sample. 597 persons, employed in five occupations with different types of work loads, rated their fatigue after work. The ratings were subjected to analyses of linear structural equation models. The results suggest a slightly revised model for perceived fatigue, still with the five dimensions: Lack of energy, Physical exertion, Physical discomfort, Lack of motivation and Sleepiness. As expected, the rating profiles describing fatigue states differed between the five occupations. On the basis of these results, a revised version of the Swedish Occupational Fatigue Inventory (SOFI) is presented. PMID- 11041306 TI - Sex differences in judgement of facial affect: a multivariate analysis of recognition errors. AB - The present paper investigated recognition errors in affective judgement of facial emotional expressions. Twenty-eight females and sixteen males participated in the study. The results showed that in both males and females emotional displays could be correctly classified, but females had a higher rate of correct classification; males were more likely to have difficulty distinguishing one emotion from another. Females rated emotions identically regardless of whether the emotion was displayed by a male or female face. Furthermore, the two-factor structure of emotion, based on a valence and an arousal dimension, was only present for female subjects. These results further extend our knowledge about gender differences in affective information processing. PMID- 11041307 TI - The interactive effect of job involvement and organizational commitment on job turnover revisited: a note on the mediating role of turnover intention. AB - This study extends previous theoretical and empirical research on Blau and Boal's (1987) model of the interactive effect of job involvement and organizational commitment on employee withdrawal. Using longitudinal data from a survey among the nursing staff of a Swedish emergency hospital (N = 535) and register information on actual turnover, the results showed, in contrast to the statement of the original theoretical model, that turnover intention mediates the additive and multiplicative effects of job involvement and organizational commitment on actual turnover. The study suggests that the proposed involvement by commitment interaction is theoretically justified, and underscores the pertinence of investigating intermediate linkages in turnover research. PMID- 11041308 TI - Descriptive features of Turkish pathological gamblers. AB - This study investigated the descriptive features of Turkish pathological gamblers. Participants were 31 male pathological gamblers and 42 "regular gamblers" who acted as controls. The subjects were diagnosed on the basis of DSM IV pathological gambling criteria and completed the Turkish Version of South Oaks Gambling Screen (SOGS). The nonpathological group was quite comparable with the pathological gambling group with respect to types and frequency of gambling and socio-demographic features. The data on the variables that defined and discriminated pathological gamblers from regular gamblers were collected through administration of a 68-item questionnaire, prepared by the authors. Compared to the non pathological gamblers, the pathological gamblers gambled more to recover their losses, experienced craving for gambling more often, gambled more often to obtain relief from disturbing emotions, harboured more irrational and unrealistic cognitions to rationalize their gambling behavior and suffered more emotionally, financially and socially as a result of their involvement in gambling. The results of the study suggested that Turkish pathological gamblers are very much like their counterparts in Western countries. PMID- 11041309 TI - A critique of recent studies on placebo effects of antidepressants: importance of research on active placebos. AB - OBJECTIVE: This review examines the recent literature on placebo effects in antidepressant drug research. Recent meta-analyses have suggested that antidepressants produce relatively small effects compared to placebo treatments, and that the effect sizes for placebo treatments are highly correlated with effect sizes for drug treatments across different studies. In addition, it is argued that "active placebos", i.e., drugs that are not antidepressants but do have discernible subjective or physiological effects, can also produce substantial changes on measures of depression. Thus, some researchers have called into question the "blind" nature of research using inert placebos, and also have questioned the efficacy of antidepressant drugs. RESULTS: The studies reviewed indicate that it is premature to suggest that antidepressant drugs have little efficacy beyond their role as active placebos. In fact, there are several important methodological, conceptual and scholarly problems with some of the work that emphasizes the role of placebo effects in mediating the actions of antidepressant drugs. CONCLUSIONS: Many of the drugs listed as "active placebos" actually have some efficacy for the treatment of depression or related disorders, and several of these drugs are used clinically to augment the effects of antidepressants. It would be important for researchers to identify new active placebos as research tools in this area, and to focus on discriminating between placebo effects and the true therapeutic actions of antidepressant drugs. A new generation of well controlled experiments in this area could shed light on these important issues, and may lead to improved methods for the determination of antidepressant efficacy. PMID- 11041310 TI - Patterns of opiate use in a heroin maintenance programme. AB - RATIONALE: Little is known about patterns of opiate use by heroin addicts. OBJECTIVES: To describe opiate use over time among heroin addicts who had access to legally prescribed intravenous heroin and oral opiates. METHODS: Analysis of daily drug administration records of 37 patients enrolled in the Geneva heroin maintenance programme for 4-29 months (total 23,136 patient-days). RESULTS: The average dose of intravenous heroin was 466 mg/day; the total opiate dose, after conversion of oral opiates to heroin-equivalents, was 543 mg/day. Patients received intravenous heroin only on 39% of days, oral opiates only on 7% of days, and mixed regimens on 49% of days; the remaining 4% of days were spent outside the programme, usually on oral opiates. The daily dose of heroin-equivalents increased during the first trimester in the programme, by 30 mg/day per month (95% confidence interval 12-46 mg/day per month), but decreased gradually thereafter, by 12 mg/day per month (95% confidence interval, 8-17 mg/day per month). In patients who alternated between heroin and methadone, 1 mg methadone typically replaced 4.1 mg heroin. During follow-up, five patients switched to methadone maintenance, five underwent detoxification, and three were discharged for noncompliance with regulations. CONCLUSIONS: Heroin users who have facilitated access to legally prescribed drugs consume about 0.5 g heroin per day. Consumption patterns vary, but the daily amount of opiates remains stable or decreases over time. A substantial minority of patients elect for alternative treatments after several months of heroin maintenance. PMID- 11041311 TI - Modulatory effects of dopamine D3/2 agonists on kappa opioid-induced antinociception and diuresis in the rat. AB - RATIONALE: The dopamine (DA) D3/2 agonist 7-OH-DPAT has been shown to attenuate the behavioral effects of the mu agonist morphine as well as the development of morphine tolerance. OBJECTIVES: To evaluate the effects of DA D3/2 agonists [7-OH DPAT, (+)-PD128,907, quinelorane, (-)-quinpirole], a D1 agonist (SKF38393), a D1 antagonist [(+)-SCH23390], a DA antagonist (spiperone), and an indirect DA agonist (cocaine) on the antinociceptive effects of kappa agonists (spiradoline, U69,593, bremazocine) as well as the effects of D3/2 agonists on the diuretic effects of spiradoline. METHODS: Antinociception was determined using a warm water (50-55 degrees C) tail-withdrawal procedure and urine output was collected over a 2-h interval. RESULTS: The antinociceptive effects produced by the kappa agonists varied with the intensity of the nociceptive stimulus (water), as maximal or near maximal effects were obtained with spiradoline at 55 degrees C, U69,593 at 52 degrees C, and bremazocine at 50 degrees C water. 7-OH-DPAT produced a dose-dependent attenuation of the antinociceptive effects of spiradoline, U69,593, and bremazocine. Spiperone completely reversed the effects of 7-OH-DPAT on spiradoline antinociception. (+)-PD128,907 and quinelorane, but not (-)-quinpirole or the other DAergic agents examined, attenuated the antinociceptive effects of spiradoline in a dose- and time-dependent manner. The diuretic effects of spiradoline were attenuated by 7-OH-DPAT, (+)-PD128,907, quinelorane, and (-)-quinpirole, and this attenuation was reversed by spiperone. CONCLUSIONS: The present study demonstrated that some D3/2 agonists can modulate both the antinociceptive and diuretic effects of kappa agonists. These modulatory actions are similar to those obtained against the effects of mu agonists. PMID- 11041312 TI - Responses to apomorphine of pigs with different coping characteristics. AB - RATIONALE: Classification of pigs based on the degree of resistance they display in a so-called "backtest" seems, to a certain extent, predictive for their coping strategy. OBJECTIVE: The present study examined whether, as found in rodents, the behavioral response to apomorphine of pigs relates to individual coping characteristics. METHODS: During the suckling period pigs were subjected to the backtest. In this test, each pig is restrained on its back for 1 min and the resistance (i.e. number of escape attempts) is scored. Pigs classified as low resisting (LR, n=10) or high-resisting (HR, n=10) were selected. At 17-18 weeks of age they received a saline and an apomorphine injection (0.2 mg/kg SC) on 2 consecutive days in a balanced design. Behavior was recorded until 120 min after injection. RESULTS: Apomorphine increased locomotion in all pigs and reduced standing, standing alert and defecating. In addition, apomorphine induced the occurrence of some peculiar activities, rarely seen in saline-treated pigs, which seemed to represent either a transition between different postures or a conflict between hind- and forelimb activities. Apomorphine-treated LR pigs performed significantly more of these activities than HR pigs. However, snout contact with the floor, an oral stereotypy, was significantly increased in apomorphine-treated HR pigs, but not in apomorphine-treated LR pigs. CONCLUSIONS: In conclusion, the response to apomorphine of pigs relates to their behavioral response, high resisting (HR) versus low-resisting (LR), in the backtest. The contrasts in behavioral response to apomorphine suggest a difference in the dopaminergic system between HR and LR pigs. PMID- 11041313 TI - Comparing the subjective, psychomotor, and physiological effects of intravenous hydromorphone and morphine in healthy volunteers. AB - RATIONALE: The psychopharmacological profile of hydromorphone, an opioid that has been used extensively for many years for post-operative pain management, has not been adequately characterized in non-drug abusers. OBJECTIVES: To characterize the subjective, psychomotor, and physiological effects of a range of single doses of hydromorphone in non-drug-abusing volunteers and to compare the effects of hydromorphone with that of morphine, a benchmark mu opioid agonist. METHODS: Subjects in a six-session study were injected in an upper extremity vein with 0, 0.33, 0.65, 1.3 mg/70 kg hydromorphone, and 5 and 10 mg/70 kg morphine, using a randomized, double-blind, crossover design. RESULTS: Hydromorphone increased scores on the pentobarbital-chlorpromazine-alcohol group and lysergic acid diethylamide scales and decreased scores on the benzedrine group scale of the Addiction Research Center Inventory, increased adjective checklist ratings of ("dry mouth", "flushing", and "nodding", and increased visual analog scale ratings indicative of both pleasant (e.g., drug liking) and unpleasant (e.g., "feel bad") effects. The subjective effects of morphine at putatively equianalgesic doses to those of hydromorphone were similar to those of hydromorphone, but in some cases of lesser magnitude. Psychomotor impairment was modest with hydromorphone and absent with morphine. Both opioids produced dose dependent decreases in pupil size. A relative potency analysis indicated that hydromorphone was 10 times as potent as morphine (1 mg hydromorphone=10 mg morphine). CONCLUSIONS: The results of this study demonstrate that 0.33-1.3 mg hydromorphone had orderly, dose-related effects on subjective, psychomotor, and physiological variables, and similar effects to those of a benchmark mu opioid agonist, morphine. PMID- 11041314 TI - Effect of hydrocortisone on the pituitary response to growth hormone releasing hormone. AB - RATIONALE: In depression, the growth hormone (GH) response to clonidine and L tryptophan (L-TRP) is reduced, suggesting reduced alpha2-adrenergic and serotonin (5-HT)1A receptor function. Pretreatment with hydrocortisone (100 mg, orally 11 h before) also blunts the GH response to L-TRP. This effect may be mediated at the hypothalamic level via reduced 5-HT1A receptor function or at the pituitary level, either by a direct effect on somatotrope cells or via enhanced insulin like growth factor-1 (IGF-1) or somatostatin (SS) release. OBJECTIVES: To examine the effects of acute and chronic exposure to hydrocortisone on baseline and stimulated GH release from the pituitary. METHODS: Twelve healthy male volunteers received pretreatment with acute hydrocortisone (100 mg, 11 h before), chronic hydrocortisone (20 mg twice a day for 1 week) and placebo in a double blind, balanced order, crossover design. Serial measurements of plasma GH, IGF-1 and thyroid stimulating hormone (TSH) levels were made at baseline and following intravenous administration of 1 mcg/kg GHRH. RESULTS: The GH response to growth hormone releasing hormone (GHRH) was significantly blunted by pretreatment with both acute and chronic hydrocortisone. Baseline IGF-1 levels were significantly lower at baseline after chronic hydrocortisone compared with placebo. Baseline TSH levels were significantly lower after acute hydrocortisone compared with placebo, suggesting an increase in somatostatin levels. CONCLUSIONS: These data suggest that hydrocortisone acts at the pituitary level to reduce GH release. The TSH and IGF-1 data support the hypothesis that hydrocortisone reduces GH release by enhancing somatostatin and IGF-1 release. PMID- 11041315 TI - Comparison of the effects of clozapine, haloperidol, chlorpromazine and d amphetamine on performance on a time-constrained progressive ratio schedule and on locomotor behaviour in the rat. AB - Performance on progressive ratio schedules has been proposed as a means of assessing the effects of drugs on "reinforcer efficacy". It has been proposed that the effects of neuroleptic drugs on operant behaviour are mediated by a reduction of "reinforcer efficacy". We examined the effects of two "conventional" neuroleptics (haloperidol and chlorpromazine) and an "atypical" neuroleptic (clozapine) on progressive ratio schedule performance; d-amphetamine was used as a comparison compound. In experiment 1, rats responded for a sucrose reinforcer on a time-constrained progressive ratio schedule (75-min sessions). After 66 preliminary training sessions, the rats received single doses (IP) of haloperidol (0.05, 0.1 mg kg(-1)). chlorpromazine (2, 4 mg kg(-1)), clozapine (0.5, 1, 2, 4, 8 mg kg(-1)), and d-amphetamine (0.2, 0.4, 0.8 mg kg(-1)), and the corresponding vehicle solutions. The highest ratio completed was reduced by haloperidol and chlorpromazine, and increased by clozapine. All three neuroleptics reduced the peak response rate, at least at the highest doses administered. Response rates on the lower and intermediate ratios could be described by a three-parameter equation proposed to account for fixed ratio schedule performance. Haloperidol reduced, and clozapine dose-dependently increased the "motivational" parameter (a); d-amphetamine reduced it at low doses and increased it at high doses. The three neuroleptics increased the "response time" parameter (delta). Un-reinforced locomotor behaviour, measured in experiment 2, was not significantly altered by haloperidol, chlorpromazine or clozapine, but was increased by d-amphetamine. These results are consistent with a reduction of reinforcer efficacy produced by haloperidol and an increase produced by clozapine; clozapine's effect is unlikely to reflect a general increase in locomotion. All three neuroleptics induced some degree of motor debilitation. The quantitative analysis of progressive ratio schedule performance may provide a useful adjunct to existing methods for separating effects of drugs on motivational and motor processes. PMID- 11041316 TI - The selective serotonin (5-HT)1A receptor ligand, S15535, displays anxiolytic like effects in the social interaction and Vogel models and suppresses dialysate levels of 5-HT in the dorsal hippocampus of freely-moving rats. A comparison with other anxiolytic agents. AB - RATIONALE: The benzodioxane, S15535, possesses low intrinsic activity and marked selectivity at 5-HT1A receptors, hippocampal populations of which are implicated in anxious states. OBJECTIVE: Herein, we examined its potential anxiolytic actions in relation to its influence upon extracellular levels of 5-HT in the dorsal hippocampus of freely-moving rats. Its effects were compared with those of other anxiolytic agents: the 5-HT1A agonists, buspirone and 8-hydroxy-2-(di-n propylamino)-tetralin HBr (8-OH-DPAT), the 5-HT2C antagonist, SB206,553 and the benzodiazepine, diazepam. METHODS: Potential anxiolytic actions were evaluated in the Vogel conflict paradigm (increase in punished responses) and the social interaction (SI) test (increase in active SI) in rats. Extracellular levels of 5 HT were determined by microdialysis. RESULTS: In analogy to diazepam. S15535 increased punished responses in the Vogel test. This action was dose dependently expressed over a broad (16-fold) dose range. Buspirone and 8-OH-DPAT were likewise active, but yielded highly biphasic dose-response curves. SB206,553 was dose dependently active in this model. In the SI test, S15535 similarly mimicked the anxiolytic-like effect of diazepam and was active over a broad dose range. Buspirone and 8-OH-DPAT again showed biphasic dose-response curves, as did SB206,553. In both the Vogel and SI tests, the anxiolytic-like effects of S15535 were abolished by the selective 5-HT1A receptor antagonist, WAY100,635, which was inactive alone. S15535 exerted its anxiolytic-like effects with a more pronounced separation to motor-disruptive doses than the other drugs. Finally, S15535 suppressed dialysate levels of 5-HT in the dorsal hippocampus, an action abolished by WAY100,635. Buspirone, 8-OH-DPAT and diazepam, but not SB206,553, also reduced 5-HT levels. CONCLUSION: Likely reflecting its distinctive ability to selectively and preferentially activate pre- versus postsynaptic 5-HT1A receptors, S15535 suppresses hippocampal 5-HT release and displays marked anxiolytic-like effects over a broad dose range in the relative absence of motor perturbation. PMID- 11041317 TI - Role of muscles accumbens dopamine D1 and D2 receptors in instrumental and Pavlovian paradigms of conditioned reward. AB - RATIONALE: This study investigated the role of nucleus accumbens dopamine D1 and D2 receptors in two different paradigms of conditioned reward. OBJECTIVE: We addressed the question whether accumbal dopamine is important for the motor or for the motivational components of reward. METHODS: We compared the effects of intra-accumbal infusion of the dopamine D1 receptor antagonist SCH23390 (0.3, 1.0, 3.0 microg) and the D2 receptor antagonist sulpiride (0.3, 1.0, 3.0 microg) on conditioned lever pressing for food, with the effects on the inhibition of the startle response by a conditioned reward signal. RESULTS: Both the D1 and the D2 antagonist dose-dependently attenuated conditioned lever pressing for reward under a fixed-ratio of responding and increased the consumption of freely available lab chow. However, the preference for freely available pellets, and the attenuation of the startle response in the presence of a conditioned stimulus predicting reward were not impaired by blockade of accumbal dopamine receptors. CONCLUSIONS: Our data support the idea that dopamine in the nucleus accumbens is necessary for instrumental response selection in the context of reward rather than for the mere motor performance of behavior or for the evaluation of the hedonic properties of rewarding stimuli. PMID- 11041318 TI - Zolmitriptan (a 5-HT1B/1D receptor agonist with central action) does not increase symptoms in obsessive compulsive disorder. AB - RATIONALE: Non-selective serotonin (5-HT) receptor agonists like meta chlorophenylpiperazine and MK-212 have been used to explore the role of 5-HT in obsessive compulsive disorder (OCD). The results of these studies and the findings of autoradiography and neuroimaging studies, pointed to a possible role of the 5-HT1B/1D receptor in the pathophysiology of OCD. Recently the selective 5 HT1B/1D receptor agonist sumatriptan was used to further explore the role of the 5-HT1B/1D receptor in OCD. Equivocal results with respect to the increase of obsessive compulsive symptoms in patients with OCD were reported. In one study a significant increase in plasma growth hormone (GH) concentration was observed, although sumatriptan does not pass the blood-brain barrier. OBJECTIVES: In order to further explore the role of the 5-HT1B/1D receptor in the pathophysiology of OCD, we performed this study, following the same design as Ho Pian et al. (Psychopharmacology 140:365-370). METHODS: In the present study we performed a randomized, double-blind, placebo-controlled, cross-over design with zolmitriptan (5 mg per os), a selective 5-HT1B/1D receptor agonist with better brain penetrating properties than sumatriptan. RESULTS: We could not detect any changes in obsessive compulsive symptoms, mood, or anxiety levels, although we found a (nonsignificant) increase in plasma GH levels. CONCLUSIONS: Based upon these findings, no evidence was found for a specific role of the 5-HT1B/1D receptor in OCD. It should be noted, however, that challenge studies in OCD are difficult to perform. Perhaps in the future better challenge paradigms will make it possible to further explore the role of specific receptor types in OCD. PMID- 11041319 TI - Long-term therapeutic drug monitoring of clozapine and metabolites in psychiatric in- and outpatients. AB - RATIONALE: Clozapine is a unique antipsychotic drug, outstanding for its lack of extrapyramidal side-effects and its superior efficacy in refractory schizophrenia. However, an unambiguous concentration-response relationship has not yet been established. OBJECTIVE: We investigated serum concentrations of clozapine, norclozapine and clozapine-N-oxide in psychiatric in- and outpatients to identify particular metabolic patterns in clozapine responders and non responders and putative threshold levels for clozapine response. METHODS: Psychiatric assessments, CYP2D6 genotype, and weekly serum concentrations of clozapine, norclozapine and clozapine-N-oxide were obtained in 34 adult schizophrenic in-and outpatients (18 men, 16 women) during 10 weeks of clozapine treatment with a naturalistic dose design. RESULTS: Responders (n=21) displayed significantly lower serum concentrations of clozapine corrected for dose compared to non-responders (n=13; P<0.05), while none of the other parameters (absolute clozapine concentration, metabolite ratios, gender) were different. Smokers had significantly lower dose-corrected clozapine concentrations. A positive correlation was observed between age and average steady state clozapine concentrations. CONCLUSIONS: These findings indicate a possible link between CYP activity and response to clozapine that is not mediated through differences in serum concentrations. No clinically meaningful pattern in serum parameters could be identified that differentiates responders from non-responders. Thus, clozapine TDM seems ineffective for predicting clinical response. Smoking behavior is a major determinant of clozapine clearance while CYP2D6 genotype does not impact clozapine disposition. PMID- 11041320 TI - Subchronic hydrocortisone treatment alters auditory evoked potentials in normal subjects. AB - RATIONALE: Abnormalities of cortical evoked potentials and background electroencephalographic (EEG) frequencies occur in several psychiatric disorders, some of which, especially depression, are associated with hypercortisolaemia. However, there have been few investigations of the effects of exogenously administered cortisol on waking EEG measures. OBJECTIVES: To examine the effects of subchronic hydrocortisone administration on auditory evoked potentials and background EEG activity. METHODS: Hydrocortisone, 20 mg twice daily, or placebo was administered to 30 normal male volunteers for 7 days in a between-subjects, double-blind trial. Auditory evoked potentials and EEG frequencies were measured on the last day. RESULTS: Hydrocortisone significantly increased the amplitudes of the N1P2 and P300 components of the auditory evoked response, but there was no change in background EEG. CONCLUSION: The results indicate that subchronic hydrocortisone treatment in normal subjects increases the amplitude of auditory evoked potentials, possibly reflecting a central alerting effect. PMID- 11041321 TI - The dopamine D3/2 agonist 7-OH-DPAT attenuates the development of morphine tolerance but not physical dependence in rats. AB - RATIONALE: The dopamine (DA) D3/2 agonist 7-OH-DPAT attenuates the acute antinociceptive, discriminative stimulus, locomotor activating, and reinforcing effects of mu agonists (for example, morphine). OBJECTIVES: To examine the ability of 7-OH-DPAT to modulate the development of morphine tolerance and physical dependence in the rat. METHODS: Morphine antinociception was assessed using a warm water tail-withdrawal procedure before and following chronic treatment with morphine (15 mg/kg)/7-OH-DPAT (0.3-3.0 mg/kg). Physical dependence was assessed following naloxone-precipitated (1.0 mg/kg) withdrawal in rats treated chronically with morphine (15 and 7.5 mg/kg)/7-OH-DPAT (1.0-10 mg/kg). RESULTS: 7-OH-DPAT attenuated the antinociceptive effects of morphine in both morphine naive and tolerant rats. Additionally, morphine tolerance was attenuated by the coadministration of 7-OH-DPAT in a dose- and time-dependent manner. The magnitude of the attenuation obtained when morphine and 7-OH-DPAT were administered at the same time was similar to that obtained when administration of these drugs was separated by 6 h, indicating that 7-OH-DPAT did not alter morphine pharmacokinetics. In rats rendered tolerant to morphine, the subsequent coadministration of morphine/7-OH-DPAT failed to reverse morphine tolerance, but did attenuate its further development. The level of physical dependence (number and frequency of withdrawal signs) was greater in rats treated with 15 than 7.5 mg/kg morphine. Under both treatment conditions, physical dependence was not altered by 7-OH-DPAT. In morphine-dependent (15 mg/kg) rats, 7-OH-DPAT (3.0 and 10 mg/kg) failed to precipitate withdrawal. CONCLUSION: The D3/2 agonist 7-OH DPAT can attenuate the antinociceptive effects of morphine in both acute and chronic preparations as well as the development of morphine tolerance. 7-OH-DPAT does not, however, alter morphine physical dependence. PMID- 11041322 TI - NKP608, an NK1 receptor antagonist, has an anxiolytic action in the social interaction test in rats. AB - RATIONALE: Evidence is starting to accumulate that NK1 receptor antagonists might have anxiolytic effects in animal tests and in patients. OBJECTIVE: To examine the effects of NKP608, a substance P antagonist acting at NK1 receptors, in various conditions of the social interaction test of anxiety and to determine its effects after 3 and 6 weeks of treatment. METHODS: Rats were tested after vehicle, 0.01 or 0.1 mg/kg PO in three conditions of the social interaction test that varied in the level of anxiety generated. Thus pairs of rats were tested in an arena with which they were unfamiliar that was lit by high (HU) or low (LU) light and in the condition that generated the lowest level of anxiety, i.e. an arena with which they were familiar, lit by low light (LF). They were also tested after 3 and 6 weeks of treatment with 0.03 mg/kg and after 24 h withdrawal from these chronic treatments. RESULTS: NKP608 had significant anxiolytic effects at 0.01, 0.03 and 0.1 mg/kg PO in the HU and LU test conditions, but was without effect in the LF condition, except for an increased incidence of bite attacks at 0.1 mg/kg. The anxiolytic effect of 0.03 mg/kg remained after 3 weeks of chronic treatment and there was no anxiogenic effect after 24 h of drug withdrawal. Following 6 weeks of chronic treatment (0.03 mg/kg per day), tolerance had developed, but no anxiogenic withdrawal effect was seen 24 h after the last dose. CONCLUSIONS: These results provide further evidence that substance P may play a role in mediating states of anxiety and suggest that the selective NK1 receptor antagonist NKP608 may prove a useful anxiolytic compound. PMID- 11041323 TI - Repeated stimulation of L-type calcium channels in the rat ventral tegmental area mimics the initiation of behavioral sensitization to cocaine. AB - RATIONALE: A substantial body of evidence indicates that ion flux through L-type calcium channels and N-methyl-D-aspartate (NMDA) receptors contributes to behavioral sensitization to cocaine. OBJECTIVES: The following experiments were designed to evaluate the role of calcium influx through L-type calcium channels or NMDA receptors in the ventral tegmental area (VTA) in the initiation of behavioral sensitization to cocaine. METHODS: The L-type calcium channel agonist BayK 8644, the glutamate agonist NMDA, or vehicle was microinjected into the VTA on 3 consecutive days. Following a 2-week withdrawal period, all rats received a challenge injection of cocaine (15 mg/kg, i.p.) in order to assess potential cross-sensitization with the NMDA or BayK 8644 pretreatments. RESULTS: Repeated intra-VTA microinjections of BayK 8644, but not NMDA, resulted in an augmentation of the behavioral response to cocaine. CONCLUSIONS: These results indicate that calcium influx through L-type calcium channels produces neurophysiological adaptations that mimic those resulting from intermittent exposure to cocaine. PMID- 11041324 TI - Acute depletion of plasma tryptophan does not alter electrophysiological variables in healthy males. PMID- 11041325 TI - Antipsychotic drugs, serum creatine kinase (CPK) and possible mechanisms. PMID- 11041326 TI - Optic nerve and chiasmal disease. AB - This review of optic nerve and chiasmal disease briefly outlines the clinical assessment and the use of diagnostic testing in the topical diagnosis of lesions of the anterior visual pathways. The commoner pathological entities including inflammatory, vascular, heredofamilial and compressive lesions are then summarised with specific reference to important points in the diagnosis and management. Specific disorders described include optic neuritis, papilloedema, ischaemic optic neuropathy, giant cell arteritis, Leber's hereditary optic neuropathy and pituitary tumours. PMID- 11041327 TI - Angiogenesis in malignant primary and metastatic brain tumors. AB - Patients with malignant primary and metastatic brain tumors have a poor prognosis, despite developments in diagnostic and therapeutic modalities. Therefore in the past decade a search for new therapeutic possibilities has started. The inhibition of angiogenesis, the sprouting of new capillaries from preexisting vasculature, which is an absolute requirement for the growth of tumors beyond a size of a few cubic millimeters, is one of the most promising approaches with which to influence tumor growth. This review focuses on the critical role of angiogenesis in the development of normal brain and the blood brain barrier. We discuss the importance of angiogenesis in the formation of malignant brain tumors and in bloodbrain barrier function in these tumors and possible consequences of altered blood-brain barrier properties for antiangiogenic therapy. Furthermore, results of current clinical trials with antiangiogenic drugs are reviewed, and clinical perspectives of antiangiogenic therapy in malignant brain tumors are outlined. PMID- 11041328 TI - Local synthesis of IgA in the cerebrospinal fluid of patients with neurological diseases. AB - To date qualitative studies of IgA in the cerebrospinal fluid in neurological disease, particularly multiple sclerosis, have been few and given mixed results. The aim of this study was to identify local synthesis of IgA by detection of clonal IgA bands, in a large cohort of patients with a variety of neurological disorders, using polyacrylamide gel electrophoresis, transfer of protein to nitrocellulose membranes and specific staining. Of 2,097 sequentially analysed patients with suspected neurological disease 54 (2.6%) had locally synthesised IgA; most notably, IgA was present in 39 of 291 (13%) patients with suspected multiple sclerosis. The latter group also had a significant excess of light-chain production, particularly free kappa, when compared to multiple sclerosis patients without local synthesis of IgA. Locally synthesised IgA was also demonstrated in inflammatory, infectious and autoimmune diseases of the central nervous system. This qualitative technique is simple and suitable for routine analysis of cerebrospinal fluid, and further qualitative studies of IgA may be useful in investigating the pathophysiology of certain neurological disorders. PMID- 11041329 TI - Incidence of CSF abnormalities in siblings of multiple sclerosis patients and unrelated controls. AB - We found that 19% (9/47) of healthy siblings of patients with clinically definite multiple sclerosis had an intrathecal immunological reaction with two or more 2 CSF-enriched oligoclonal bands (OCBs), in contrast to (4%) (2/50) unrelated healthy controls. Furthermore, in this group of nine healthy sibs the measles CSF IgG antibody titers were higher than that of the other sibs and that of controls. There were also differences in the serum titers for measles IgG antibody, which were higher in the group of all healthy sibs than in healthy volunteers, and (as with CSF titers) higher in the subgroup of healthy sibs with two or more 2 CSF enriched OCBs than the other sibs. Thus a significant proportion of healthy siblings to MS patients have a partially hyperimmune condition similar to that occurring in MS, which in 19% manifested itself as an OCB reaction, in 9% as increased CSF measles IgG antibody titers, and in 21% as increased serum measles IgG antibody titers, these phenomena tending to occur in the same individuals. This condition is characterized by CSF-enriched OCBs with undefined specificity, although some increased antiviral reactivity is found both in the serum and CSF. While it needs further characterization, a genetic trait interacting with common infections is suggested. The recurrence risk of this condition is approximately five times higher than the 3-4% recurrence risk for manifest MS reported for sibs. PMID- 11041330 TI - Identification of the spinocerebellar ataxia type 7 mutation in Taiwan: application of PCR-based Southern blot. AB - Spinocerebellar ataxia (SCA) type 7 is an autosomal dominant disorder characterized by neural loss, mainly in the cerebellum and regions of the brainstem and particularly the inferior olivary complex. This neurodegeneration disease is associated with expansion of unstable CAG repeats within the 5' translated region of the SCA7 gene, located on chromosome 3p. We conducted a local survey of the normal population and candidate patients for the analysis of the CAG repeats in the SCA7 gene. The distributions of the CAG repeat units of SCA7 gene in the normal population in Taiwan were established in this study by using the radioactive genomic polymerase chain reaction (PCR). The normal range of CAG repeats is from 6 to 17 repeats, with the more common being around 8-13 repeats. The range is narrower than that reported for other ethnic groups (7-35 CAGs). Meanwhile, by the use of a combination of PCR and Southern blot analysis, one SCA7 family was identified and is reported here. A marked instability of the CAG repeat number during transmission from father to son (41 vs. 100) was observed in the SCA7 family. Clinical anticipation is significant in this family including an infantile case, who was found to have nystagmus from the age of 1 month. To date, the SCA7 mutation has been detected in one of 73 families with autosomal dominant cerebellar ataxia phenotypes, which is about 1.4% of the ataxia families referred to us, compared to 1.4% SCA1, 9.6% SCA2, and 27.3% SCA3/Machado-Joseph disease in our collection. In addition, we demonstrate that the PCR-based Southern blot analysis, with the advantages of sensitivity of PCR and specificity of Southern blot, is a reliable diagnostic method for SCA7 mutation screening. The molecular analysis technique makes possible the quick and accurate diagnosis of SCA7 patients and in the future will hopefully be applied to prenatal screening for SCA7 families. PMID- 11041331 TI - The sternocleidomastoid test: an in vivo assay to investigate botulinum toxin antibody formation in humans. AB - In a small number of patients treated with botulinum toxin (BT) antibody (Ab) formation occurs. BT Ab can be detected by the mouse protection assay (MPA) or by the mouse diaphragm assay (MDA). Both methods, however, have major drawbacks. We tested a method for detecting BT Ab which measures the BT-induced reduction in the electromyographic amplitude of the mean maximal voluntary activation (M-EMG) of the sternocleidomastoid muscle. The M-EMG reduction was compared in 17 patients with cervical dystonia and secondary BT therapy failure to the M-EMG reduction previously measured in controls. Values more than 2 SD below the mean of controls were considered abnormal. Six patients showed BT Ab on the MPA and MDA; all of these had abnormal M-EMG reductions. Eleven patients showed no BT Ab on MPA and MDA testing; in ten of these the M-EMG reduction was normal, and in one it was pathological, but MDA testing later changed to positive under continued BT therapy. The sternocleidomastoid test is easy to perform and produces quantitative results. Since its sensitivity and specificity are at least as good as those of the MDA and the MPA, it can replace them. PMID- 11041332 TI - Increased amyloid beta protein in the skin of patients with amyotrophic lateral sclerosis. AB - Distinct vascular and periadnexal immunoreactivity have been observed for amyloid b protein (Abeta) in skin from patients with amyotrophic lateral sclerosis (ALS). We used an enzyme-linked immunosorbent assay to make a more quantitative comparison of Abeta concentrations between ALS patients and controls. The insoluble fractions of skin samples from ALS patients contained significantly higher Abeta concentrations per milligram protein than those from controls. Various alterations in extracellular matrix components have been reported to occur in the skin of patients with ALS, and several matrix constituents have been shown to affect processing and aggregation of Abeta in human brain. Taking these previous findings together with those of the present study, our observations suggest that changes in extracellular matrix in skin of ALS patients may facilitate aggregation and deposition of Abeta. PMID- 11041333 TI - Difference in pathogenesis between herpes simplex virus type 1 encephalitis and tick-borne encephalitis demonstrated by means of cerebrospinal fluid markers of glial and neuronal destruction. AB - We determined the extent of neuronal and glial cell destruction in 13 patients with herpes simplex type 1 (HSV-1) encephalitis, 15 patients with tick-borne encephalitis (TBE), and 20 noninfectious controls by analyzing the cerebrospinal fluid (CSF) concentrations of neurofilament protein (a marker of neurons, mainly axons), neuron-specific enolase (a marker of neurons, mainly somas), glial fibrillary acidic protein, and S-100 protein (markers of astrocytes). In addition, in patients with HSV-1 encephalitis CSF samples were collected serially before 7, 8-14, and 18-49 days and 3-10 months after the onset of neurological symptoms. In the acute stage of HSV-1 encephalitis we found markedly higher CSF levels of the cell damage markers than in patients with TBE. The concentration of cell damage markers in HSV-1 encephalitis decreased within 45 days after acute infection, except for neurofilament protein. The CSF concentrations of neurofilament protein increased during the second week, remained extremely high throughout the next month, and decrease thereafter. The changes in these markers of neuronal and glial destruction demonstrate the neuronal and astroglial cell damage during the first month after HSV-1 encephalitis. In contrast, most patients with TBE had signs only of slight astrogliosis, except for two patients with paresis. PMID- 11041334 TI - Epilepsy, Brugada syndrome and the risk of sudden unexpected death. PMID- 11041335 TI - Non-systemic vasculitic neuropathy presenting with a painful polyradiculopathy: a case report. PMID- 11041336 TI - Mandibular drop resulting from bilateral metastatic trigeminal neuropathy as the presenting symptom of lung cancer. PMID- 11041337 TI - Simultanagnosia in a patient with right brain lesions. PMID- 11041338 TI - Unilateral ophthalmoparesis and limb ataxia associated with anti-GQ1b IgG antibody. PMID- 11041339 TI - Slowly progressive spinal muscular atrophy of the hands (O'Sullivan-McLeod syndrome): clinical and magnetic resonance imaging presentation. PMID- 11041340 TI - Neoplastic cells in the cerebrospinal fluid in intravascular lymphomatosis. PMID- 11041341 TI - Leon Rostan (1790-1866). PMID- 11041342 TI - Crucial role of heparan sulfate proteoglycan (agrin) in beta-amyloid formation in Alzheimer's disease. PMID- 11041343 TI - Pyoverdines: pigments, siderophores and potential taxonomic markers of fluorescent Pseudomonas species. AB - Pyoverdine, the yellow-green, water-soluble, fluorescent pigment of the fluorescent Pseudomonas species, is a powerful iron(III) scavenger and an efficient iron(III) transporter. As a fluorescent pigment, it represents a ready marker for bacterial differentiation and, as a siderophore, it plays an important physiological function in satisfying the absolute iron requirement of these strictly aerobic bacteria. Close to 40 structurally different pyoverdines have been identified to date, each characterized by a different peptidic part of the molecule and by a very narrow specificity as an iron transporter for Pseudomonas species, usually restricted to the producer strain or to strains producing an identical compound. Cross-reactivity does occur, however, for pyoverdines exhibiting partial identity at the peptide chain level, suggesting some information on the receptor-recognition site of the molecule. With the recent description of an operonic cluster of four genes involved in the synthesis of the chromophoric part of the molecule, a total of seven pyoverdine biosynthetic genes have been identified so far in Pseudomonas aeruginosa PAO1. Although the precise function of the gene products needs further clarification, a biosynthetic pathway based on a multienzyme thiotemplate mechanism allowing a step-by-step synthesis of the whole chromopeptide molecule can be postulated. A promising future is expected from recent developments which indicate that pyoverdines might be considered as potent and easy-to-handle taxonomic markers for the fluorescent species of the genus Pseudomonas. PMID- 11041344 TI - Deletion of the hmc operon of Desulfovibrio vulgaris subsp. vulgaris Hildenborough hampers hydrogen metabolism and low-redox-potential niche establishment. AB - The hmc operon of Desulfovibrio vulgaris subsp. vulgaris Hildenborough encodes a transmembrane redox protein complex (the Hmc complex) that has been proposed to catalyze electron transport linking periplasmic hydrogen oxidation to cytoplasmic sulfate reduction. We have replaced a 5-kb DNA fragment containing most of the hmc operon by the cat gene. The resulting chloramphenicol-resistant mutant D. vulgaris H801 grows normally when lactate or pyruvate serve as electron donors for sulfate reduction. Growth with hydrogen as electron donor for sulfate reduction (acetate and CO2 as the carbon source) is impaired. These results confirm the importance of the Hmc complex in electron transport from hydrogen to sulfate. Mutant H801 is also deficient in low-redox-potential niche establishment. On plates, colony development takes 14 days longer than colony development of the wild-type strain, when the cells use hydrogen as the electron donor. This result suggests that, in addition to transmembrane electron transport from hydrogen to sulfate, the redox reactions catalyzed by the Hmc complex are crucial in establishment of the required low-redox-potential niche that allows single cells to grow into colonies. PMID- 11041345 TI - Characterization of novel bacteriochlorophyll-a-containing red filaments from alkaline hot springs in Yellowstone National Park. AB - Novel red, filamentous, gliding bacteria formed deep red layers in several alkaline hot springs in Yellowstone National Park. Filaments contained densely layered intracellular membranes and bacteriochlorophyll a. The in vivo absorption spectrum of the red layer filaments was distinct from other phototrophs, with unusual bacteriochlorophyll a signature peaks in the near-infrared (IR) region (807 nm and 911 nm). These absorption peaks were similar to the wavelengths penetrating to the red layer of the mats as measured with in situ spectroradiometry. The filaments also demonstrated maximal photosynthetic uptake of radiolabeled carbon sources at these wavelengths. The red layer filaments displayed anoxygenic photoheterotrophy, as evidenced by the specific incorporation of acetate, not bicarbonate, and by the absence of oxygen production. Photoheterotrophy was unaffected by sulfide and oxygen, but was diminished by high-intensity visible light. Near-IR radiation supported photoheterotrophy. Morphologically and spectrally similar filaments were observed in several springs in Yellowstone National Park, including Octopus Spring. Taken together, these data suggest that the red layer filaments are most similar to the photoheterotroph, Heliothrix oregonensis. Notable differences include mat position and coloration, absorption spectra, and prominent intracellular membranes. PMID- 11041346 TI - Purification, properties and primary structure of alanine dehydrogenase involved in taurine metabolism in the anaerobe Bilophila wadsworthia. AB - Alanine dehydrogenase [L-alanine:NAD+ oxidoreductase (deaminating), EC 1.4.1.4.] catalyses the reversible oxidative deamination of L-alanine to pyruvate and, in the anaerobic bacterium Bilophila wadsworthia RZATAU, it is involved in the degradation of taurine (2-aminoethanesulfonate). The enzyme regenerates the amino group acceptor pyruvate, which is consumed during the transamination of taurine and liberates ammonia, which is one of the degradation end products. Alanine dehydrogenase seems to be induced during growth with taurine. The enzyme was purified about 24-fold to apparent homogeneity in a three-step purification. SDS PAGE revealed a single protein band with a molecular mass of 42 kDa. The apparent molecular mass of the native enzyme was 273 kDa, as determined by gel filtration chromatography, suggesting a homo-hexameric structure. The N-terminal amino acid sequence was determined. The pH optimum was pH 9.0 for reductive amination of pyruvate and pH 9.0-11.5 for oxidative deamination of alanine. The apparent Km values for alanine, NAD+, pyruvate, ammonia and NADH were 1.6, 0.15, 1.1, 31 and 0.04 mM, respectively. The alanine dehydrogenase gene was sequenced. The deduced amino acid sequence corresponded to a size of 39.9 kDa and was very similar to that of the alanine dehydrogenase from Bacillus subtilis. PMID- 11041347 TI - Seasonal and spatial community dynamics in the meromictic Lake Cadagno. AB - The seasonal and spatial variations in the community structure of bacterioplankton in the meromictic alpine Lake Cadagno were examined by temporal temperature gradient gel electrophoresis (TTGE) of PCR-amplified 16S rDNA fragments. Two different amplifications were performed, one specific for the domain Bacteria (Escherichia coli positions 8-536) and another specific for the family Chromatiaceae (E. coli positions 8-1005). The latter was followed by semi nested reamplification with the bacterial primer set, allowing comparison of the two PCR approaches by TTGE. The TTGE patterns of samples from the chemocline and the anoxic monimolimnion were essentially identical, whereas the oxic mixolimnion displayed distinctively different banding patterns. For samples from the chemocline and the monimolimnion, dominant bands in the Bacteria-specific TTGE profiles comigrated with bands obtained by the semi-nested PCR approach specific for Chromatiaceae. This observation suggested that Chromatiaceae are in high abundance in the anoxic water layer. All dominant bands were excised and sequenced. Changes in the community structure, as indicated by changes in the TTGE profiles, were observed in samples taken at different times of the year. In the chemocline, Chomatium okenii was dominant in the summer months, whereas Amoebobacter purpureus populations dominated in autumn and winter. This change was confirmed by fluorescent in situ hybridization. PMID- 11041348 TI - Mercury uptake and removal by Euglena gracilis. AB - The uptake and removal of mercury (added as HgCl2) from the culture medium by Euglena gracilis was studied. In cultures initiated in the light, cells accumulated a small fraction of the added heavy metal (5-13%). Mercury was both biologically and nonbiologically volatilized, and cell growth was partially inhibited; under these conditions the glutathione content was 3.2 nmol/10(6) cells. In contrast, in cultures initiated in the dark, mercury uptake by cells was two to three times higher, biological volatilization remained unchanged and nonbiological volatilization and growth were negligible; the glutathione content diminished to 1.4 nmol/10(6) cells. Biological mercury volatilization depended on cell density and metal concentration, but was light-independent. Thus, volatilization of mercury by Euglena appeared not to be an effective mechanism of resistance, whereas a high intracellular level of glutathione and a low mercury uptake seemed necessary for successful tolerance. PMID- 11041349 TI - Ultrastructure of Acaryochloris marina, an oxyphotobacterium containing mainly chlorophyll d. AB - We present a detailed investigation of the ultrastructure of the chlorophyll a/d containing unicellular oxyphotobacterium Acaryochloris marina, combining light and transmission electron microscopy and showing freeze fractures of this organism for the first time. The cells were 1.8-2.1 microm x 1.5-1.7 microm in size. The cell envelope consisted of a peptidoglycan layer of approximately 10 nm thickness combined with an outer membrane. Cell division was intermediate between the constrictive and the septum type. The nucleoplasm, which contained several carboxysomes, was surrounded by 7-11 concentrically arranged thylakoids, which were predominantly stacked, with the exception of distinct areas where phycobiliproteins were located. The thylakoids were perforated by channel-like structures connecting the central and peripheral portions of the cytoplasm and not yet observed in other organisms. In freeze fractures, the protoplasmic fracture faces of thylakoid membranes were densely covered with particles of inhomogenous size. The particle size histogram peaked at 10-11, 13 and 18 nm. The 18-nm particles are assumed to represent photosystem I trimers. The particles on exoplasmic fracture faces, proposed to represent photosystem II complexes, were significantly larger than the corresponding particles of cyanobacteria and clustered to form large aggregates. This kind of arrangement is unique among photosynthetic organisms. PMID- 11041350 TI - Fermentation of 4-aminobutyrate by Clostridium aminobutyricum: cloning of two genes involved in the formation and dehydration of 4-hydroxybutyryl-CoA. AB - Clostridium aminobutyricum ferments 4-aminobutyrate via succinic semialdehyde, 4 hydroxybutyrate, 4-hydroxybutyryl-CoA and crotonyl-CoA to acetate and butyrate. The genes coding for the enzymes that catalyse the interconversion of these intermediates are arranged in the order abfD (4-hydroxybutyryl-CoA dehydratase), abfT (4-hydroxybutyrate CoA-transferase), and abfH (NAD-dependent 4 hydroxybutyrate dehydrogenase). The genes abfD and abfT were cloned, sequenced and expressed as active enzymes in Escherichia coli. Hence the insertion of the [4Fe-4S]clusters and FAD into the dehydratase required no additional specific protein from C. aminobutyricum. The amino acid sequences of the dehydratase and the CoA-transferase revealed close relationships to proteins deduced from the genomes of Clostridium difficile, Porphyromonas gingivalis and Archaeoglobus fulgidus. In addition the N-terminal part of the dehydratase is related to those of a family of FAD-containing mono-oxygenases from bacteria. The putative assignment in the databank of Cat2 (OrfZ) from Clostridium kluyveri as 4 hydroxybutyrate CoA-transferase, which is thought to be involved in the reductive pathway from succinate to butyrate, was confirmed by sequence comparison with AbfT (57% identity). Furthermore, an acetyl-CoA:4-hydroxybutyrate CoA-transferase activity could be detected in cell-free extracts of C. kluyveri. In contrast to glutaconate CoA-transferase from Acidaminococcus fermentans, mutation studies suggested that the glutamate residue of the motive EXG, which is conserved in many homologues of AbfT, does not form a CoA-ester during catalysis. PMID- 11041351 TI - A selDABC cluster for selenocysteine incorporation in Eubacterium acidaminophilum. AB - The four genes required for selenocysteine incorporation were isolated from the gram-positive, amino acid-fermenting anaerobe Eubacterium acidaminophilum, which expresses various selenoproteins of different functions. The sel genes were located in an unique organization on a continuous fragment of genomic DNA in the order selD1 (selenophosphate synthetase 1), selA (selenocysteine synthase), selB (selenocysteine-specific elongation factor), and selC (selenocysteine-specific tRNA). A second gene copy, encoding selenophosphate synthetase 2 (selD2), was present on a separate fragment of genomic DNA. SelD1 and SelD2 were only 62.9% identical, but the two encoding genes, selD1 and selD2, contained an in-frame UGA codon encoding selenocysteine, which corresponds to Cys-17 of Escherichia coli SelD. The function of selA, selB, and selC from E. acidaminophilum was investigated by complementation of the respective E. coli deletion mutant strains and determined as the benzyl viologen-dependent formate dehydrogenase activity in these strains after anaerobic growth in the presence of formate. selA and selC from E. acidaminophilum were functional and complemented the respective mutant strains to 83% (selA) and 57% (selC) compared to a wild-type strain harboring the same plasmid. Complementation of the E. coli selB mutant was only observed when both selB and selC from E. acidaminophilum were present. Under these conditions, the specific activity of formate dehydrogenase was 55% of that of the wild type. Transformation of this selB mutant with selB alone was not sufficient to restore formate dehydrogenase activity. PMID- 11041352 TI - Protection of Methanosarcina barkeri against oxidative stress: identification and characterization of an iron superoxide dismutase. AB - Methanosarcina barkeri is a methanogenic archaeon that can only grow under strictly anoxic conditions but which can survive oxidative stress. We have recently reported that the organism contains a monofunctional catalase. We describe here that it also possesses an active iron superoxide dismutase. The enzyme was purified in three steps over 130-fold in a 14% yield to a specific activity of 1500 U/mg. SDS-PAGE revealed the presence of only one band, at an apparent molecular mass of 25 kDa. The primary structure determined from the cloned and sequenced gene revealed similarity to iron- and manganese superoxide dismutases. The highest similarity was to the iron superoxide dismutase from Methanobacterium thermoautotrophicum. The enzyme from M. barkeri was found to contain, per mol, 1 mol iron, but no manganese in agreement with the general observation that anaerobically growing organisms only contain iron superoxide dismutase. The enzyme was not inhibited by cyanide (10 mM), which is a property shared by all iron- and manganese superoxide dismutases. The presence of superoxide dismutase in M. barkeri is noteworthy since a gene encoding superoxide dismutase (sod) has not been found in Archaeoglobus fulgidus, a sulfate-reducing archaeon most closely related to the Methanosarcinaceae. PMID- 11041353 TI - Dual control of melanogenesis and melanoma growth: overview molecular to clinical level and the reverse. AB - Utilizing increased melanin pigmentation and accentuated melanogenesis seen in malignant melanoma, we newly developed melanoma-selective boron neutron capture therapy (BNCT) after designing and synthesizing the 10B-DOPA analogue, 10B-p boronophenylalanine (10B-BPA). After multi-disciplined and extensive basic and pre-clinical investigations, we successfully treated 18 cases of human melanoma. Recently, we found that accentuated synthesis of melanin monomers, richest within coated vesicles (CV) in melanoma cells, plays a critical role in attracting 10B BPA through chemical complex formation of monomers and 10B-BPA. CV are indeed BPA localizing organelles. This led us to the new clinical endeavor that BPA may possess the potential ability to suppress melanin polymer formation through 'melanin monomer trapping' out of the melanogenic pathway which is highly regulated by the function of CV in pigment cells. It was soon found that melanin polymer formation can be suppressed by BPA at the chemical and cellular levels, then at the clinical level. Our discovery, that single molecule 10B-BPA possesses the dual nature of eradication of melanoma with BNCT and suppression of melanin hyperpigmentation, resulted from pursuing bilateral feedback at each stage from pure science to clinical application and vice versa. A further example of bilateral feedback is the development of gene-transfer applied BNCT (gBNCT). This also has its roots in clinical hurdles faced in treating amelanotic melanomas by 10B-BPA BNCT. The transfer of tyrosinase and melanin monomer synthesis-related genes into target cancer cells has produced more effective BNCT and may lead to gBNCT for non-melanoma cancers. PMID- 11041354 TI - The melanosome: the perfect model for cellular responses to the environment. AB - Melanocytes play critical roles in mammals, including the regulation of constitutive pigmentation in the skin, hair, and eyes, in embryological development, and in photoprotection from ionizing radiation. The pigments themselves, possibly due to the inherent cytotoxic properties of their precursors, are synthesized and deposited within membrane-bound organelles known as melanosomes. However, the structure of melanosomes, and thus their characteristic properties, varies widely, from relatively disorganized, poorly pigmented pheomelanosomes to highly structured, melanized eumelanosomes. Melanocytes respond to various physiological stimuli, such as melanocyte stimulating hormone (MSH), agouti signal protein (ASP), endothelins and/or ultraviolet light (UVL) by highly complex intracellular signaling mechanisms that can elicit dramatic changes in melanosome and melanocyte morphology. MSH and UVL stimulate transcription of melanogenic genes that elicit dramatic increases in the amount of eumelanins produced, whereas ASP serves as an antagonist of MSH and inhibits the transcription of those same genes. Recent studies have shown that melanocyte-specific transcription factors, such as MITF, play important roles in these responses, but ubiquitous transcription factors, such as ITF2 and E2A, are also involved. Virtually all known intracellular signaling pathways affect one or more parameters of pigmentation, and it is clear that both melanocyte-specific and basic housekeeping processes are affected by such modulation. The properties of melanins, including their photoprotective function, may be optimized by such stimulatory responses. Studies targeted at elucidating the regulatory mechanisms involved and the functional changes that result demonstrate that the melanosome is the perfect model to study such biological response mechanisms. PMID- 11041355 TI - Prolactin signaling in erythrophores and xanthophores of teleost fish. AB - Prolactin directly affects erythrophores and xanthophores of teleost fish, resulting in pigment dispersion. In the present study, signal transduction elicited by prolactin was examined using split-tail fin preparations of the rose bitterling and Nile tilapia, and cultured erythrophores and xanthophores from the paradise goby and rose bitterling. When antibodies to the prolactin receptor were added to an ovine prolactin (oPRL) solution, pigment dispersion within cultured cells was significantly inhibited, suggesting the existence of a prolactin receptor in the cell membrane. In mammals and birds, prolactin receptors belong to a cytokine receptor superfamily and signal through a tyrosine kinase-mediated pathway. Therefore, we examined the effects of three kinds of protein tyrosine kinase inhibitors on pigment dispersion elicited by oPRL. None of those inhibitors depressed the response. On the other hand, lithium ions (an inhibitor of adenylate cyclase) and H-88 and H-89 (inhibitors of protein kinase A) decreased the levels of oPRL-induced pigment dispersion in a dose-dependent manner. In cultured cells treated with cholera toxin for 3 hrs, the effect of oPRL was irreversible, indicating the possible involvement of Gs protein in the prolactin action. From these results, we conclude that cAMP may be a second messenger in the dispersion of pigment induced by prolactin and that a novel protein receptor coupled with a Gs protein may be present in the membrane of erythrophores and xanthophores of teleost fish. PMID- 11041356 TI - Intrinsic and extrinsic pathomechanisms in vitiligo. AB - Vitiligo is the most commonly acquired hypomelanosis, and is restricted to a limited cutaneous territory (focal/segmental vitiligo [SV]) or generalized in symmetric patches (nonsegmental vitiligo [NSV]). In the majority of cases, vitiligo corresponds to a loss of melanocytes, first in the epidermal compartment, and later in the follicular reservoir where most melanocytic stem cells are probably situated. There are many data currently supporting an impaired redox status of the epidermal melanin unit as a primary defect leading to inappropriate immune responses in NSV. SV is probably a mosaic developmental predisposition to melanocytic loss, with similar mechanisms at work on a limited scale, as suggested by its cutaneous distribution and success of autografting. In NSV, engraftment of autologous melanocytes is less durable, especially in areas prone to repeated trauma or pressure. Although melanocytes are the obvious target of the disease, keratinocytes, as providers of antioxidant molecules to melanocytes as well as cofactors in the synthesis of melanin, are probably involved. The production of autoantibodies and specific cytotoxic T cells is not surprising in the context of the massive uptake of melanocytic antigens by Langerhans cells in unstable vitiligo vulgaris, thereby allowing the self perpetuation of lesions. This article reviews the recent data on the pathophysiology of vitiligo, on the basis of clinical classification and the intrinsic/extrinsic nature of proposed pathomechanisms. Unfortunately, basic issues like pathological staging, clinical scoring, and eliciting factors have not yet been fully resolved. More genetic studies in vitiligo-prone families and in specific genetic disorders associated with vitiligo are also needed. PMID- 11041357 TI - Biochemical and genetic studies of pigment-type switching. AB - Mutations that affect the balance between the synthesis of eumelanin and pheomelanin provide a powerful set of tools with which to understand general aspects of cell signaling. Previous work from our laboratory has demonstrated that pheomelanin synthesis is triggered by the ability of Agouti protein to inhibit signaling through the Melanocortin 1 receptor (Mc1r). In a bioassay based on the Xenopus Mc1r, Agouti protein has two effects, competitive inhibition of receptor occupancy by alpha-MSH and down-regulation of receptor signaling, which are mediated separately by domains in the amino- and carboxy-terminal regions of Agouti protein, respectively. Recently, we have used the genetics of pigmentation as an in vivo system to screen for and analyze other mutations in the Agouti melanocortin pathway. The pigmentary effects of Agouti are suppressed by the previously existing coat-color mutations mahogany (mg), mahoganoid (md), and Umbrous (U). Double mutant studies, with animals deficient for the Mc1r or those which carry Ay, indicate that mg and md are genetically upstream of the Mc1r, and can suppress the effects of Ay on both pigmentation and body weight. Positional cloning has recently identified the gene mutated in mahogany as a single transmembrane-spanning protein whose ectodomain is orthologous to human Attractin (Atrn). PMID- 11041358 TI - Insect pigmentation: activities of beta-alanyldopamine synthase in wing color patterns of wild-type and melanic mutant swallowtail butterfly Papilio glaucus. AB - Color pattern formation was studied in wild-type and melanic swallowtails because of their unique pigment system, the papiliochromes, which are derived from the tyrosine as well as from the tryptophan pathway. In a comparative approach we used females of Papilio glaucus which occur in two phenotypes, either wild-type (yellow and black) or melanic. Pigment synthesis in the developing wings starts with formation of yellow papiliochromes followed later by black melanin. From earlier studies we know that dopamine produced from DOPA by the enzyme dopadecarboxylase (DDC), is a precursor of both black melanin and also of N-beta alanyldopamine (NBAD) in yellow papiliochrome synthesis. Thus, DDC expression and enzyme activity is required in both types of pigment forming scale cells and occurs in a time and pattern specific manner. However, differential activity of DDC alone can not be sufficient to regulate synthesis of different pigments in differently colored scales. Therefore, we tested the hypothesis whether activity of another enzyme, beta-alanyldopamine synthase (BAS), regulates specifically papiliochrome synthesis. BAS transfers beta-alanine to dopamine to give NBAD a component of yellow papiliochrome. We developed a radio-enzyme-assay of BAS activity in which (14C)-beta-alanine is incubated with dopamine, Mg++-ions and ATP together with wing homogenates containing putative BAS activity. In fact, high BAS activity was measured in yellow wings in concert with a high DDC activity. In contrast, in melanic wings almost no BAS activity was found. From this result it is clear, that papiliochrome synthesis in yellow scales is switched on by BAS shifting dopamine into the papiliochrome pathway and out of the melanin pathway or vice versa. PMID- 11041359 TI - Abnormal vesicular trafficking in mouse models of Hermansky-Pudlak syndrome. AB - Hermansky-Pudlak Syndrome (HPS) is a group of related multigenic recessively inherited disorders which causes abnormalities in the biosynthesis and/or function of three related organelles; melanosomes, platelet-dense granules and lysosomes. These lead, in turn, to hypopigmentation, prolonged bleeding and ceroid deposition. Positional cloning strategies have identified five mouse HPS genes. Two orthologous human diseases (HPS1 and HPS2) have likewise been identified. At least four of the five mouse genes encode proteins involved in the regulation of intracellular vesicle trafficking. The pearl (HPS2) and mocha genes encode the beta3A and delta subunits, respectively, of the AP-3 adaptor complex, which captures organelle membrane proteins at the trans-Golgi apparatus. The protein products of the pallid and gunmetal genes are also important components of the vesicular trafficking machinery. The former interacts with a t-SNARE, syntaxin13, and the latter is the alpha subunit of Rab geranylgeranyltransferase, which renders Rab proteins sufficiently lipophilic to function at their target membranes. The pale ear (HPS1) gene encodes a ubiquitously expressed protein of unknown function. Recent physiological studies have shown that mouse HPS mutants, like their human HPS counterparts, have variably reduced lifespans and may have lung abnormalities. PMID- 11041360 TI - Oestrogenic steroids and melanoma cell interaction with adjacent skin cells influence invasion of melanoma cells in vitro. AB - The invasion of melanoma is complex and multi-staged and involves changes in both cell/extracellular matrix (ECM) and cell/cell interactions. Female steroids and alpha-MSH have also been reported to influence metastatic melanoma progression, but their mechanisms of action are unknown. Accordingly, our aim was to establish in vitro models to examine (a) the influence of sex steroids and alpha-melanocyte stimulating hormone (alpha-MSH) on tumour invasion and the influence of (b) ECM proteins and (c) adjacent cells on melanoma invasion. In the first model, melanoma cell invasion through fibronectin over 20 hr under serum-free conditions was used to investigate the effects of 17beta-oestradiol and oestrone on the invasion of human melanoma cell lines, A375-SM and HBL. A375-SM, but not HBL cells, proved very susceptible to inhibition by female steroids. However, invasion of the HBL line was inhibited by alpha-MSH. Using the second model of reconstructed human skin based on de-epidermised acellular dermis, we found that the HBL cells on their own failed to invade into the dermis (irrespective of the presence or absence of the basement membrane). However, there was a significant synergistic interaction between keratinocytes, fibroblasts and HBL cells, such that a modest invasion of HBLs into the dermis was seen within 2 weeks when other skin cells were present. In contrast, A375-SM cells showed a significant ability to invade the dermis in the absence of other cells, with less invasion when other skin cells were present. In summary, these models have provided new information on the extent to which melanoma cell invasion is sensitive to oestrogenic steroids and to alpha-MSH and to interaction, not only with adjacent skin cells but also to the presence of basement membrane antigens. PMID- 11041361 TI - Stem cell factor and/or endothelin-3 dependent immortal melanoblast and melanocyte populations derived from mouse neural crest cells. AB - Stem cell factor (SCF) and endothelin-3 (ET3) are both necessary for melanocyte development. In order to obtain immortal cell populations of melanoblasts that can survive without feeder cells, we first obtained an immortal cell population of neural crest cells (NCCs) from Sl/+ and +/+ mice of strain WB by incubating with a culture medium supplemented with SCF and ET3, and then we designated them as NCC-SE3 cells. NCC-SE3 cells were bipolar, polygonal, or round in shape and possessed melanosomes of stages I-III (mainly stage I). They were positive to dihydroxyphenylalanine (DOPA) reaction and expressed KIT (a receptor tyrosine kinase), tyrosinase, tyrosinase-related protein-1 (TRP1), tyrosinase-related protein-2 (TRP2), and endothelin-B receptor (ETRB) as determined by immunostaining. We next cultured NCC-SE3 cells by changing culture medium from the one supplemented with SCF + ET3 to the one supplemented with SCF or ET3. NCC SE3 cells cultured with ET3 alone, designated as NCC-E3 cells, were bipolar in shape and had mainly stage II melanosomes and expressed the same proteins as did NCC-SE3 cells. However, NCC-SE3 cells cultured with SCF alone, designated as NCC S4.1 cells, were polygonal in shape and had mainly stage I melanosomes. They are thought to be more immature because they were positive to KIT, TRP1, and TRP2, but not to ETR(B), tyrosinase, and DOPA reaction. When 12-O-tetradecanoylphorbol 13-acetate and cholera toxin were added to the culture medium, NCC-S4.1 cells changed shape from polygonal to bipolar and became DOPA-positive. This suggests that NCC-S4.1 cells are melanoblasts that have the potential to differentiate into melanocytes. These cell populations will be extremely useful to study factors that affect melanocyte development and melanogenesis. PMID- 11041362 TI - Regulation of growth and melanogenesis of uveal melanocytes. AB - We have developed methods for the isolation, cultivation, and investigation of human uveal melanocytes (UM). Uveal melanocytes grow well and produce melanin in vitro in the presence of basic fibroblast growth factor (bFGF), cyclic adenosine monophosphate-elevating agents, and serum. Cultured UM respond to various factors. Certain growth factors (bFGF and hepatocyte growth factor, etc.), endothelin, adrenergic beta2-receptor agonists, and some prostaglandins (EP2 receptor agonists and certain TP-receptor agonists) stimulate, while transforming growth factor-beta2, interleukin-6, and cholinergic agonists inhibit melanogenesis and/or growth of UM in vitro. Alpha-melanocyte-stimulating hormone, adrenocorticotropic hormone, various sex hormones, and prostaglandin F2alpha showed no effect on the growth and melanogenesis of cultured UM. The stability of UM in vivo may be controlled by these factors. Disturbance of this balance may lead to certain rare pathologic pigmentary changes of the iris. UM are relatively stable in vivo; they usually do not respond (proliferate or show dynamic changes in melanogenesis) to various environmental factors. The differences of the in vivo behavior between uveal and epidermal melanocytes may be determined by both cellular factors and environmental factors. PMID- 11041363 TI - An antisense strategy for inhibition of human melanoma growth targets the growth factor pleiotrophin. AB - A major biological characteristic of metastatic melanomas is their ability to survive in a growth-factor-depleted environment, whereas normal melanocytes die rapidly under such conditions. The increased survival of melanoma cells is due to their production of growth factors for autocrine growth stimulation. Here, we describe a strategy to inhibit pleiotrophin (PTN), a heparin-binding autocrine growth factor for melanoma cells. To target PTN production in melanoma cells, a replication-deficient, recombinant adenovirus was generated to express antisense (AS) PTN at high efficiency. The AS vector induced transcripts that completely inhibited PTN protein production. Melanoma growth was strongly inhibited if the tumor cells were maintained in a three-dimensional environment in soft agar, whereas cell growth was not affected if the tumor cells were grown as a monolayer, suggesting the importance of cell-matrix interactions for the biological activity of this growth factor. The down-regulation of PTN in transduced melanoma cells coincided with the down-regulation of the cell-cycle regulator cyclin E and the up-regulation of the cell-cycle inhibitor p21WAF1/Cip1. Tumor growth in vivo was also delayed by the AS-PTN vector, confirming that PTN is essential for the three-dimensional growth of tumor cells. Our studies demonstrate the importance of assessing potential melanoma antagonists not only on cells grown as monolayers but also in three-dimensional matrices. PMID- 11041364 TI - Tanning as part of the eukaryotic SOS response. AB - We have determined that DNA damage is at least one of the signals generated by ultraviolet radiation that stimulates pigmentation (tanning) in human skin. This photoprotective response is functionally similar to the SOS response described in bacteria. Here we present evidence that DNA damage stimulates pigmentation, at least in part, through up-regulation of tyrosinase mRNA and protein levels. Furthermore, this response can be induced in the absence of DNA damage by treatment of melanocytic cells and intact skin with small DNA fragments, particularly thymidine dinucleotides, pTpT. Topical application of these DNA fragments should provide a photoprotective tan to human skin without the harmful effects of ultraviolet radiation. PMID- 11041365 TI - Regulation of pigment cell-specific gene expression by MITF. AB - Melanocyte and retinal pigment epithelium (RPE) specifically express tyrosinase and other melanin-producing enzymes. Microphthalmia-associated transcription factor (Mitf), encoded at the mouse microphthalmia locus, regulates the development and survival of many cell types, including melanocyte and RPE. MITF, the human homolog of Mitf, consists of at least four isoforms with distinct amino termini, referred to as A, MITF-C, MITF-H, and MITF-M. MITF-M is exclusively expressed in melanocytes and melanoma cells, and thus represents the melanocyte lineage-specific isoform. In contrast, other isoforms are expressed in many cell types so far examined. These isoforms appear to function as transcriptional activators of the melanogenesis genes, as assessed by transient transfection assays in cultured cells. Functional significance of Mitf-M in melanocyte differentiation was verified by the molecular lesion of black-eyed white Mitf(mi bw) mice, which lack melanocytes but have normal RPE. The Mitf gene of this mutant has the insertion of an L1 retrotransposable element in the intron between exon 3 and exon 4, leading to complete repression of Mitf-M mRNA expression. Taken together, these results suggest that melanogenesis in melanocyte and RPE is regulated by separate Mitf/MITF isoforms. Recent findings on the multiplicity of MITF isoforms are summarized. PMID- 11041366 TI - Chemical analysis of melanins and its application to the study of the regulation of melanogenesis. AB - Melanins are difficult to characterize because of their intractable chemical properties, the heterogeneity in their structural features, and the lack of methods to split melanin polymers into monomer units. To overcome this difficulty, we developed a rapid and sensitive method for quantitatively analyzing eumelanin and pheomelanin in biological samples that is based on the formation of pyrrole-2,3,5-tricarboxylic acid and/or aminohydroxyphenylalanine followed by HPLC determination. The method has been applied to the study of melanogenesis. The results summarized in this review are: 1) Biochemical studies show that in the process of mixed melanogenesis, cysteinyldopas are produced first, which are then oxidized to give pheomelanin; following cysteine depletion, eumelanin is then deposited on the preformed pheomelanin. 2) In vitro and in vivo studies show that tyrosinase activity is the most important factor that regulates the switch of melanogenesis, with lower tyrosinase activities favoring pheomelanogenesis; further suppression of melanogenesis results in a lack of pigment production. 3) In cultured melanocytes, the concentrations of tyrosine and cysteine, and their ratio in the medium, are important in determining the concentrations of eumelanin and pheomelanin produced and their ratio in the cells. In conclusion, our HPLC microanalytical method for characterizing eumelanin and pheomelanin has become a useful tool for the study of melanogenesis. PMID- 11041367 TI - Intracellular vesicular trafficking of tyrosinase gene family protein in eu- and pheomelanosome biogenesis. AB - The intracellular vesicular trafficking in the melanosome biogenesis (melanogenesis) is reviewed with the incorporation of our own experimental findings. The melanosome biogenesis involves four stages of melanosome maturation, which reflect the transport of structural and enzymatic proteins from Golgi (trans-Golgi network: TGN) to the melanosomal compartment and their organization therein. The major melanosomal proteins include tyrosinase gene family protein (tyrosinase and tyrosinase-related protein; TRP), lysosome associated membrane protein (Lamp) and gp100 (pmel 17). They are glycosylated in the endoplasmic reticulum, and transported by vesicles from the TGN to the melanosomal compartment. During the formation of transport vesicles, they assemble on the cytoplasmic face of the TGN to select cargo by interacting directly or indirectly with coat proteins. Tyrosinase and TRP-1 possess the dileucine motifs at the cytoplasmic domain, to which adapter protein-3 binds to transport them from the TGN to stage I melanosomes (related to late endosomes) and then to stage II melanosomes. A number of small guanosine triphosphate binding proteins, including rab 7, appear to be involved in this vesicular transport. Phosphatidyl inositol 3 kinase also regulates this membrane trafficking of melanosomal glycoprotein. Eumelanogenesis is controlled by melanocyte-stimulating hormone, and all three tyrosinase gene family proteins are transported from the TGN to stage II melanosomes that are elliposoidal and contain the structural matrix of filaments/lamellae. In contrast, pheomelanogenesis is primarily regulated by agouti signal protein, and only tyrosinase is transported from stage I melanosomes to stage II melanosomes that are spherical and related to lysosomes. Because of the absence of TRP-1 and TRP-2 in pheomelanogenesis, it may be suggested that tyrosinase is involved in lysosomal degradation after forming dopaquinone, to which the cysteine present in the lysosomal granule binds to form cysteinyldopas that will then be auto oxidized to become pheomelanin. PMID- 11041368 TI - New insights on the structure of the mouse silver locus and on the function of the silver protein. AB - The melanosomal proteins encoded by the silver locus play important roles in melanogenesis. The human locus yields two proteins, PMEL17 and GP100, by alternative mRNA splicing. The mouse si locus was reported to encode a Pmel17 protein, and later gp87, a GP100 homologue. When we re-examined the products of wild-type and silver-mutant mouse si loci, RT-PCR of wild-type RNA and genomic DNA sequence accounted for gp87 but excluded the occurrence of Pmel17. Analysis of cDNA from the silver (si/si) melanocyte line, melan-si, showed that the pathogenic mutation is a G to A substitution at nt 1808, which yields a premature stop codon and a predicted protein truncated in the C-terminus. This was confirmed by reaction of a specific anti-gp87 antiserum with si/si melanocyte extracts. To further explore gp87 function, we compared the DHICA oxidase activity of extracts from B16, melan-si (heterozygous for the brown mutation and homozygous for the silver mutation) and Cloudman S91 cells (homozygous for the brown mutation), since both TRP1 and gp87 are thought to be involved in DHICA oxidation/polymerization. Cloudman extracts do not oxidize significantly DHICA and its methyl ester, supporting the involvement of native mouse TRP1 in DHICA oxidation. Extracts from B16 and melan-si do not show significant differences for the oxidation of free acid and methylated dihydroxyindoles, indicating that the mechanism is not decarboxylative. Melan-si extracts are very efficient in catalyzing dihydroxyindole oxidation, in spite of being heterozygous for the TRP1 mutation, consistent with a stablin effect for the wild-type gp87 protein. On the other hand, aggregated and degraded forms of that mutant gp87 protein are found in the cytosolic fraction of melan-si, suggesting that misrouting and aberrant processing of the gp87 and tyrosinase may also be related to the high DHICA oxidase activity of these melanocytes. PMID- 11041369 TI - Skin POMC peptides: their actions at the human MC-1 receptor and roles in the tanning response. AB - The melanocortin 1 (MC-1) receptor is a key control point in the regulation of skin pigmentation. Alpha-MSH is an agonist at this receptor and through its activation regulates melanocyte function. alpha-MSH is cleaved from pro opiomelanocortin (POMC) in the pituitary, but in humans the skin is a more important source of the peptide. Skin pigmentation is therefore regulated by locally produced alpha-MSH rather than that of pituitary origin. alpha-MSH acts as a paracrine and/or autocrine mediator of UV induced pigmentation. However, the predominant alpha-MSH in human skin is desacetyl alpha-MSH and, compared to the acetylated form, is a relatively weak agonist at the human MC-1 receptor. By acting as a partial agonist desacetyl alpha-MSH may even oppose the actions of acetylated alpha-MSH and other MC-1 receptor agonists. The most abundant MC-1 receptor agonist in human epidermis is ACTH1-17. This POMC peptide, which is produced by keratinocytes, is more potent than acetylated alpha-MSH in stimulating melanogenesis in human melanocytes and, in contrast to the latter, produces a biphasic dose-response curve. This is probably a consequence of its activation of both the cAMP and IP3/DAG signalling pathways. alpha-MSH peptides, on the other hand, selectively activate the cAMP pathway. Compared with alpha MSH, ACTH1-17 could have the more important role as a paracrine mediator of melanogenesis and other melanocytic processes. However, ACTH1-17 is not the only POMC peptide in the skin and may interact with related peptides at the MC-1 receptor. These interactions are likely to represent important determinants of melanocyte function and skin pigmentation. PMID- 11041370 TI - Molecular bases of congenital hypopigmentary disorders in humans and oculocutaneous albinism 1 in Japan. AB - The molecular bases of various types of congenital hypopigmentary disorders have been clarified in the past 10 years. Homozygous gene mutations of enzymes functional in melanogenesis such as tyrosinase, P protein and DHICA oxidase, result in oculocutaneous albinism (OCA) 1, OCA 2, and OCA 3, respectively. The genes responsible for Hermansky-Pudlak syndrome (HPS) and Chediak-Higashi syndrome (CHS) have also recently been isolated and cloned. The transcription factor paired box 3 (PAX3) works at the promoter region of the microphthalmia associated transcription factor (MITF) gene, and the MITF transcription factor orders the expression of c-kit, which encodes the receptor for stem-cell factor, which in turn stimulates melanoblast migration from the neural tube to the skin in the embryo. Heterozygous mutations of PAX3, MITF, or c-kit genes induce Waardenburg syndrome (WS) 1/3, WS 2 or Piebaldism, respectively. A defect of endothelin-3 or the endothelin-B receptor produces WS 4. In our examination of 26 OCA 1 patients in Japan, all were found to have homozygous or heterozygous tyrosinase gene mutations at codons 77 or 310. Therefore, mutations at codons 77 and 310 are the major ones in Japanese patients with OCA 1. An autosomal dominant pigmentary disease of dyschromatosis symmetrica hereditaria (DSH) is well known in Japan, and is characterized by a mixture of hypo- and hyper-pigmented macules of various sizes on the backs of the hands and feet. The disease gene and its chromosomal localization have not been identified yet. Our trial of linkage analysis and positional cloning to determine the disease gene is presented. PMID- 11041371 TI - Malignant melanoma on the sole: how to detect the early lesions efficiently. AB - Early detection of malignant melanoma (MM) is essential to improve the prognosis. In non-white populations, including Japanese, the sole is the most prevalent site of MM. On the sole, however, melanocytic nevus is also frequently found. Clinical differentiation of early MM from benign melanocytic nevus on the sole is sometimes difficult because both are observed as a brownish-black macule. For the effective early detection of MM on the sole, the author has proposed guidelines based on the data of hundreds of melanocytic lesions on the sole. The algorithmic guidelines are as follows: when you see a pigmented lesion on the sole, first exclude congenital melanocytic nevus and some other specified disorders, and then measure the maximum diameter of the lesion. If it is more than 7 mm, biopsy it for histopathologic evaluation. If it is 7 mm or less, just follow the course of the lesion and advise the patient to come back if it enlarges to more than 7 mm. Even when the lesion is 7 mm or less, a biopsy is recommended on it, if it shows marked irregularity in shape and/or color or it shows the parallel ridge pattern with epiluminescence microscopy (ELM). The author believes the guidelines surely work efficiently in screening early MM on the sole. PMID- 11041372 TI - UVA, pheomelanin and the carcinogenesis of melanoma. AB - Cloudman S91 mouse melanoma cells vary in constitutive and inducible melanin levels. Survival, mutation induction and DNA damage were quantitated after exposure to UVB, UVA and FS20 lamps. Assuming that the observed differences are related to melanin, induced pigment is photo-protective for survival and mutation after UVB and FS20 exposure, and is photosensitizing for survival after UVA exposure. No changes in pyrimidine dimers could be measured. DNA damage in pigmented mouse melanocytes (melan-a and melan-b) was greater than that in albino melanocytes (melan-c) after UVB and FS20, and the pigmented cells were more sensitive to killing. Pigment appears to be protective for killing by UVA in these melanocytes. Human melanocytes from different skin types vary in both melanin amount and composition (eu- and pheomelanin). Effects of pigmentation on UVB responses are unclear. In UVA, heavily pigmented cells have more DNA damage than lightly pigmented cells, but are resistant to killing. Increased pheomelanin photosensitizes DNA damage in lightly pigmented cells. Since eumelanin predominates in the mouse melanoma cells and melanocytes, they are less likely than human cells to provide a satisfactory model for human solar melanomagenesis. In order to understand the mechanism of photocarcinogenesis of melanoma, melanins in human melanocytes from different pigment types should be carefully quantitated and characterized. Mutations induced in them by solar wavelength-emitting lamps with well-characterized spectra should be measured, and mutant DNA should be sequenced to determine the nature of the solar-induced lesions. Research should focus on UVA and pheomelanin. PMID- 11041373 TI - Microphthalmia: a signal responsive transcriptional regulator in development. AB - The transcription factor Microphthalmia (Mi) is a helix-loop-helix protein which plays an essential role in the development and subsequent function of neural crest-derived melanocytes. Since its discovery as a mutant allele producing mice devoid of viable melanocytes, Mi has emerged as the gene whose mutation is responsible for the human pigmentation condition Waardenburg Syndrome IIa, as well as a variety of cellular defects involving retinal pigment epithelium, osteoclasts, and mast cells. As discussed here, Mi has recently been recognized to be targeted by several signaling pathways of importance to melanocytes, those activated by melanocyte stimulating hormone (MSH) and Steel factor. While these two pathways profoundly modulate Mi activity, they do so via strikingly different mechanisms. The differences and similarities of these responses highlight the likely roles of Mi in influencing both pigmentation and proliferation/survival. These effects are also of significance in human melanoma, a tumor for which Mi appears to be an extremely sensitive and specific marker. PMID- 11041374 TI - Evolution and development of pigment cells: at the crossroads of the discipline. AB - The following is a summary of the current state of comparative biology with respect to pigmentation. Recent results from molecular analyses of genes involved in pigmentation in lower vertebrates are compared with similar data from mouse and man. Particular emphasis has been placed on evolutionary and developmental aspects of pigmentation. Recent advances in molecular biology of lower vertebrate pigmentation allow for the comparison of orthologous molecules across a wider range of species than ever before; some of these results are summarized and used to highlight the current state of pigmentation from a comparative perspective. A more cellular, organismal approach is also explored to highlight some important lessons from comparative biology. Lastly, large-scale evolutionary questions are put into a framework that highlights both the differences and similarities between mammals/birds and other vertebrates. It is the opinion of the authors that important, long-standing questions in these areas can now be addressed in ways that have not been possible before. Thus, the discipline is at an exciting crossroads where developmental and evolutionary data can be used to create a unified view of pigment cells and pigments across many species. PMID- 11041375 TI - The melanocortin-1 receptor is a key regulator of human cutaneous pigmentation. AB - The cloning and characterization of the human melanocortin-1 receptor (MC1R) and the demonstration that normal human melanocytes respond to the melanocortins, alpha-melanocyte stimulating hormone (alpha-MSH) and adrenocorticotrophic hormone (ACTH), with increased proliferation and eumelanogenesis had put an end to a long standing controversy about the role of melanocortins in regulating human cutaneous pigmentation. We have shown that alpha-MSH and ACTH bind the human MC1R with equal affinity, and are equipotent in their mitogenic and melanogenic effects on human melanocytes. We also showed that the activation of the MC1R is important for the melanogenic response of human melanocytes to ultraviolet radiation (UVR). The MC1R is also the principal mediator of the inhibitory effects of agouti signaling protein (ASP) on melanogenesis. Expression of the MC1R is subject to regulation by its own ligands alpha-MSH and ACTH, as well as by UVR and endothelin-1. Recent studies that we conducted on the expression of MC1R variants by human melanocytes and the implications of these variants on the function of the MC1R revealed the following. Human melanocytes homozygous for Arg160Trp mutation in the MC1R demonstrated a significantly reduced response to alpha-MSH. Also, this culture responded poorly to ASP and exhibited an exaggerated cytotoxic response to UVR. Another culture, which was homozygous for Val92Met mutation in the MC1R, demonstrated a normal response to alpha-MSH. Heterozygous mutations that are frequently expressed in various melanocyte cultures did not disrupt MC1R function. These results begin to elucidate the significance of MC1R variants in the function of the receptor. Our data emphasize the significance of a normally functioning MC1R in the response of melanocytes to melanocortins, ASP, and UVR. PMID- 11041376 TI - T cell immune responses against melanoma and melanocytes in cancer and autoimmunity. AB - T cell responses specific for melanoma cells and melanocytes appear to be involved in the rejection of melanoma tumors, as well as in the development of autoimmune reactions in patients with Vogt-Koyanagi-Harada disease (VKH), sympathetic ophthalmia, or autoimmune vitiligo. Some of the target antigens for those T cells have been isolated using cDNA expression cloning with melanoma reactive T cells derived from lymphocytes tumor infiltrating (TIL) of patients with melanoma. These include melanocyte specific proteins, such as tyrosinase, TRP1, TRP2, gp100, and MART-1, cancer-testis antigens, and mutated peptides derived from genetic alterations in melanoma cells. Some of the melanoma reactive T cells appear to respond to cryptic or subdominant self epitopes in melanosomal proteins. Modification of those epitopes to increase their immunogenicity by replacement of amino acids at primary anchor residues for peptide/MHC binding, allowed an improvement in immunotherapy for patients with melanoma. Targets for autoreactive T cells against melanocytes in those autoimmune disorders remain to be identified. Isolation of novel target antigens is important for understanding these pathological T cell responses, as well as for developing new diagnostic and treatment methods for these diseases. A variety of techniques, including cDNA expression cloning with T cells, serological analysis of recombinant cDNA expression libraries (SEREX), cDNA subtraction with representational differential analysis (RDA), and serial analysis of gene expression (SAGE) are now being applied to identify novel melanoma/melanocyte antigens recognized by T cells and antibodies. PMID- 11041377 TI - Depigmenting effect of alpha-tocopheryl ferulate on normal human melanocytes. AB - Oral vitamin E supplementation has been reported to improve facial hyperpigmentation. alpha-Tocopheryl ferulate (alpha-TF) is a compound of alpha tocopherol (alpha-T) and ferulic acid connected by an ester bond. Ferulic acid is also an antioxidant, and could scavenge free radicals induced by ultraviolet (UV) radiation, and thus maintain the long-lasting antioxidative effect of alpha-T. Previously we have reported that alpha-TF inhibited melanogenesis in human melanoma cells. To know whether alpha-TF might be useful as a whitening agent to improve and prevent facial hyperpigmentation, the depigmenting effect of alpha-TF in normal human melanocytes was examined in this study. The results showed that 30 microg/ml of alpha-TF dissolved in 150 microg/ml of lecithin inhibited melanization significantly without inhibiting cell growth. This phenotypic change was associated with the inhibition of tyrosinase and the degree of inhibition was dose dependent. No significant effect on DOPAchrome tautomerase (DT) activity was observed. These results suggest that alpha-TF is a candidate for an efficient whitening agent which suppresses melanogenesis. In this paper, the role of alpha T and alpha-TF in inhibiting biological reactions induced by reactive oxygen species (ROS) is also discussed. PMID- 11041378 TI - History, evolution, and diagnosis of premenstrual dysphoric disorder. AB - Clinically significant premenstrual problems with mood and behavior have been recognized since ancient times. However, it was not until 1987 that formal criteria for a specific diagnosis were proposed. The history of the development of the DSM-IV criteria for premenstrual dysphoric disorder, as well as the epidemiology and ways in which the condition differs from other mood disorders, is reviewed. PMID- 11041379 TI - Recognizing and treating premenstrual dysphoric disorder in the obstetric, gynecologic, and primary care practices. AB - The author's aim is to aid primary care physicians and obstetrician-gynecologists in correctly diagnosing and treating premenstrual dysphoric disorder (PMDD). The symptoms fluctuate markedly, but their timing is key. PMDD patients experience symptoms only during the luteal phase and will have a symptom-free interval after the menstrual flow and before ovulation. The author discusses self-report instruments, which are valuable tools for diagnosis when combined with the ICD-10 criteria for premenstrual syndrome (PMS) or the DSM-IV criteria for PMDD and the ruling out of medical and psychiatric conditions, such as diabetes, hypothyroidism, major depression, and dysthymia, that cause similar symptoms. Treatment strategies ranging from nonpharmacologic approaches such as dietary modification and aerobic exercise to pharmacologic interventions such as antidepressants, anxiolytics, and agents to suppress ovulation are examined. PMID- 11041380 TI - Premenstrual dysphoria and the serotonin system: pathophysiology and treatment. AB - The inclusion of research diagnostic criteria for premenstrual dysphoric disorder (PMDD) in the DSM-IV recognizes the fact that some women have extremely distressing emotional and behavioral symptoms premenstrually. PMDD can be differentiated from premenstrual syndrome (PMS), which presents with milder physical symptoms, headache, and more minor mood changes. In addition, PMDD can be differentiated from premenstrual magnification of physical and/or psychological symptoms of a concurrent psychiatric and/or medical disorder. As many as 75% of women with regular menstrual cycles experience some symptoms of PMS, according to epidemiologic surveys. PMDD is much less common; it affects only 3% to 8% of women in this group. The etiology of PMDD is largely unknown, but the current consensus is that normal ovarian function (rather than hormone imbalance) is the cyclical trigger for PMDD-related biochemical events within the central nervous system and other target organs. The serotonergic system is in close reciprocal relationship with the gonadal hormones and has been identified as the most plausible target for interventions. Thus, beyond the conservative treatment options such as lifestyle and stress management, other nonantidepressant treatments, or the more extreme interventions that eliminate ovulation altogether, the serotonin reuptake inhibitors (SRIs) are emerging as the most effective treatment option for this population. Results from several randomized, placebo-controlled trials in women with PMDD have clearly demonstrated that the SRIs have excellent efficacy and minimal side effects. More recently, several preliminary studies indicate that intermittent (premenstrual only) treatment with selective SRIs is equally effective in these women and, thus, may offer an attractive treatment option for a disorder that is itself intermittent. PMID- 11041381 TI - Non-antidepressant treatment of premenstrual syndrome. AB - Although selective serotonin reuptake inhibitors are considered the first-line treatment option for premenstrual syndrome, several other such options are also available. Multiple studies have indicated that medications that suppress ovulation alleviate premenstrual emotional and physical symptoms. However. the use of such medications, such as the gonadotropin-releasing hormone agonists, leads to prolonged low estrogen levels and cardiac and osteoporotic health risks. A recent double-blind, placebo-controlled study of 466 women with premenstrual syndrome reported that calcium was effective in reducing emotional, behavioral, and physical premenstrual symptoms. Recent preliminary trials have suggested efficacy for cognitive therapy, light therapy, and tryptophan. Future studies of diet recommendations, exercise, relaxation, magnesium, nonsteroidal anti inflammatory drugs, diuretics, opiate antagonists, and alternative therapies are needed. PMID- 11041382 TI - Auditory and vestibular mouse mutants: models for human deafness. AB - We have shown here several examples of how hearing and vestibular impaired mouse mutants are generated and the insight that they provide in the study of auditory and vestibular function. These types of genetic studies may also lead to the identification of disease-susceptibility genes, perhaps the most critical element in presbyacusis (age-related hearing loss). Some individuals may be more prone to hearing loss with increasing age or upon exposure to severe noise, and susceptibility genes may be involved. Different inbred mice show a variety of age related and noise-induced hearing loss that varies between normal hearing and severe deafness throughout their life span /27/. Genetic diversity between inbred mouse strains has been shown to be a powerful tool for the discovery of modifier genes. Already two studies have found regions in which modifier genes for deafness may reside /28-29/. Future studies will hopefully lead to the identification of genes that modify hearing loss and will help us understand the variability that exists in human hearing, a crucial component in developing successful treatment strategies. The first human non-syndromic deafness-causing gene was identified in 1995, and since then, additional genes have been discovered. Much of the credit for this boom is due to deaf and vestibular mouse mutants. Their study has led to great insight regarding the development and function of the mammalian inner ear, and correlations with human deafness can now be made since mutations in the same genes have been found in these two mammals. As deafness is the most common form of sensory impairment and affects individuals of all ages, elucidating the function of the auditory and vestibular systems through genetic approaches is essential in improving and designing effective treatments for hearing loss. PMID- 11041383 TI - Use of evoked potentials to objectively differentiate between selective vulnerability of cochlear and vestibular end organ function. AB - Auditory nerve brainstem evoked responses (ABR) have been used for several decades to investigate cochlear function. Recently techniques have been developed to elicit similar recordings from the vestibular end organs - short latency vestibular evoked potentials (VsEPs). Both ABR and VsEP reflect appropriate end organ function and may therefore be used to investigate the vulnerability of these end organs to various experimental insults, such as noise exposure and ototoxic drugs. PMID- 11041384 TI - Evaluating children's hearing by DPOAEs at 1-10 kHz. AB - The efficacy of application of DPOAE (distortion product otoacoustic emission) in the evaluation of hearing in children was assessed. 106 children were checked by both DPOAE and ABR. The results show that DPOAE is highly effective in discriminating between hearing disorders and normal hearing. The correct classification rate of normal ears varied between 86.8-96.8% and ears with hearing loss (ABR threshold equal or above 20 dB HL) 87-100%. At the lower frequencies (1-2 kHz) the hit rate was lower. The results of this study suggest that the DPOAE can be used as a rapid hearing screening test for infants and children providing frequency specific information, mostly in the 3-10 kHz frequency range. The effectiveness of DPOAE at 1-10 kHz was evident in children with middle ear dysfunction and in detecting infants with possible high frequency hearing loss. In addition, high frequency DPOAE broadens the range typically available from click-evoked ABR measurements. This information is of increased clinical value not only in terms of speech and language development but also for detecting children who are at high-risk for possible hearing deterioration and who require early intervention. PMID- 11041385 TI - The varieties of auditory neuropathy. AB - Auditory neuropathy (AN) was initially described as impairment of auditory neural function, with preserved cochlear hair cell function. In this report, 67 patients with audiological and neurophysiological criteria for hearing loss due to auditory neuropathy are described. Reviewing this large body of patients, AN appears to consist of a number of varieties, with different etiologies and sites affected. All varieties share a relatively spared receptor function, and an impaired neural response, with diminished ability to follow fast temporal changes in the stimulus, but different varieties in this general scheme can be distinguished. Analyses of the clinical features indicate that auditory neuropathies vary in several measures including age of onset, presence of peripheral neuropathy, etiology, and behavioral and physiological measures of auditory function. The sites affected along the peripheral auditory pathway may include dysfunction of the outer hair cells, the synapse between hair cell and auditory nerve, and the auditory nerve fibers, with myelin as well as axonal impairments contributing to the disorder. PMID- 11041386 TI - Objective detection and localization of multiple sclerosis lesions on magnetic resonance brainstem images: validation with auditory evoked potentials. AB - To develop an objective method for detecting multiple sclerosis (MS) brainstem lesions, magnetic resonance (MR) images (multiple planar, spin-echo, acquired in three planes of section) of sixteen MS patients and fourteen normal subjects were analyzed with an algorithm that detected regions with a relatively increased intensity on both a spin-echo image and a T2 image. To be considered a lesion, such regions had to overlap in at least two orthogonal planes. Using a digitized atlas of the human brainstem, the lesion locations were mapped with respect to the brainstem anatomy. This method was evaluated by comparing the location of MS lesions with the brainstem auditory evoked potentials obtained from these subjects. Brainstem lesions were detected in five MS patients; four had lesions impinging upon the auditory system and one did not. All four had abnormal evoked potentials. The fourteen normal subjects, the one MS patient with brainstem lesions outside the auditory pathway, and the eleven other MS patients with no brainstem lesions all had normal evoked potentials. The requirement that lesions be detected in at least two planes of section greatly improved the specificity of the algorithm. The consistency between the MR and brainstem auditory evoked potentials results supports the validity of this imaging analysis algorithm for objectively localizing brainstem lesions. PMID- 11041387 TI - Identification of the relative onset time of a two-tone complex: auditory sensitivity or language based? AB - Categorical perception occurs when equal physical differences on a continuum sometimes cause no changes in the identification of a stimulus, while at other points on the same continuum, that difference causes an abrupt change in perception. One of the unresolved issues regarding this phenomenon is what determines categorical perception: a special speech mode or "natural" psychophysical boundaries. One way to answer this question is by investigating categorical perception with non-speech stimuli. An example is the identification of the relative onset of a two-tone complex (TOT) analogous to voice-onset time (VOT), the acoustic cue to voicing in initial position. Studies in English found similar category boundaries for TOT and VOT (at +20 ms) supporting the non-speech specific theory. The purpose of this study was to investigate TOT in Hebrew speaking listeners whose language uses very different VOT values from those reported in English. Twenty Hebrew-speaking young adults participated in this study. Stimuli consisted of a two-tone complex that varied in the relative onset time of the lower tone from a lead of -50 ms to a lag of +50 ms in 10 ms steps. Results show that: (1) All subjects were able to identify the lag conditions from the simultaneous ones but only half of them were also able to identify the lead from the simultaneous ones. This was explained in terms of prominent pitch cues available when shifting from simultaneous to lagging stimuli. (2) Hebrew category boundaries (CBs) for TOT are shorter than those of VOT, and both are shorter than the respective English ones. Nonetheless, all CBs fell into the range of 10 to 30 ms. The data support the hypothesis that "natural" psychophysical boundaries determine categorical perception but behavioral measures may be influenced by speech. PMID- 11041388 TI - Separation of conjoined twins. PMID- 11041389 TI - Parents' and GPs' key role in diagnosis of meningococcal septicaemia. PMID- 11041390 TI - BSE and transmission through blood. PMID- 11041391 TI - Surgery for colorectal cancer in elderly patients. PMID- 11041392 TI - Non-invasive ventilation for chronic obstructive pulmonary disease. PMID- 11041393 TI - Is syndrome "X" another late complication of bone-marrow transplantation? [comment]. PMID- 11041394 TI - Dopamine and schizophrenia--proof at last? PMID- 11041395 TI - The less acceptable face of bias. PMID- 11041396 TI - Recombinant bactericidal/permeability-increasing protein (rBPI21) as adjunctive treatment for children with severe meningococcal sepsis: a randomised trial. rBPI21 Meningococcal Sepsis Study Group. AB - BACKGROUND: Endotoxin is a primary trigger of the inflammatory processes that lead to shock, multiorgan failure, and purpura fulminans in meningococcal sepsis. Bactericidal/permeability-increasing protein (BPI) is a natural protein, stored within the neutrophil granules, that binds to and neutralises the effects of endotoxin in vitro, in laboratory animals, and in humans. To establish whether a recombinant 21-kDa modified fragment of human BPI (rBPI21), containing the active antimicrobial and endotoxin-neutralising moiety, would decrease death and long term disability from meningococcal sepsis, we did a randomised, double-blind, placebo-controlled trial of rBPI21 in children with severe meningococcal sepsis. METHODS: We enrolled children (2 weeks to 18 years of age) presenting to 22 centres in the UK and the USA with a clinical picture suggestive of meningococcal sepsis, and with evidence of severe disease. Children were randomly assigned rBPI21 (2 mg/kg over 30 min followed by 2 mg/kg over 24 h) or placebo (0.2 mg/mL human albumin solution) in addition to conventional medical therapy. Primary outcome variables were mortality, amputations, and change in paediatric overall performance category (POPC) from before illness to day 60. Analysis was by intention to treat. FINDINGS: Of 1287 patients screened, 892 were excluded, including 57 patients who died or who met criteria for imminent death before receiving the study drug. 190 patients received rBPI21, and 203 placebo. 34 (8.7%) of 393 patients died during the study: 14 (7.4%) in the rBPI21 group and 20 (9.9%) in the placebo group (odds ratio 1.31 [95% CI 0.62-2.74], p=0.48). Compared with patients randomised to placebo, fewer patients treated with rBPI21 had multiple severe amputations (six of 190 [3.2%] vs 15 of 203 [7.4%], odds ratio 2.47 [0.94-6.51], p=0.067), and more had a functional outcome similar to that before illness (as measured by the POPC scale) at day 60 (136 of 176 [77.3%] vs 126 of 190 [66.3%], p=0.019). INTERPRETATION: Because most deaths occurred in the interval between identification of patients and study drug administration, the mortality rate in the placebo group was substantially lower than predicted. The trial was therefore underpowered to detect significant differences in mortality. However, patients receiving rBPI21 had a trend towards improved outcome in all primary outcome variables. Given the excellent severity match between placebo and rBPI21 groups at study entry, the results overall indicate that rBPI21 is beneficial in decreasing complications of meningococcal disease. PMID- 11041397 TI - Surgery for colorectal cancer in elderly patients: a systematic review. Colorectal Cancer Collaborative Group. AB - BACKGROUND: The effectiveness of surgery for colorectal cancer depends on it being carried out safely, which allows most patients to return to productive lives, with an improved postoperative life expectancy, or at least one that is not diminished by the surgery. Because colorectal cancer is a major cause of morbidity and mortality in elderly people, we have examined how the outcomes of surgery in elderly patients differ from those in younger patients. METHODS: We did a systematic review of published and aggregate data provided by investigators. Studies were identified by computerised and manual searches of published and unpublished reports, scanning references, and contacting investigators. Within each study, outcomes for patients aged 65-74 years, 75-84 years, and 85+ years were expressed in relation to those aged less than 65 years. FINDINGS: From 28 independent studies, and a total of 34,194 patients, we found that elderly patients had an increased frequency of comorbid conditions, were more likely to present with later-stage disease and undergo emergency surgery, and less likely to have curative surgery than younger patients. The incidence of postoperative morbidity and mortality increased progressively with advancing age. Overall survival was reduced in elderly patients, but for cancer specific survival age-related differences were much less striking. INTERPRETATION: The relation between age and outcomes from colorectal cancer surgery is complex and may be confounded by differences in stage at presentation, tumour site, pre existing comorbidities, and type of treatment received. However, selected elderly patients benefit from surgery since a large proportion survive for 2 or more years, irrespective of their age. PMID- 11041398 TI - Risk factors for arrhythmia and sudden cardiac death late after repair of tetralogy of Fallot: a multicentre study. AB - BACKGROUND: Ventricular arrhythmia and sudden cardiac death late after repair of tetralogy of Fallot are devastating complications in adult survivors of early surgery, but their prediction remains difficult. METHODS: We examined surgical, electrocardiographic, and late haemodynamic data, and their relation to clinical arrhythmia and sudden death occurring over 10 years, in a multicentre cohort of patients with repaired tetralogy, who were alive in 1985. RESULTS: Of 793 patients (mean age at repair 8.2 years [SD 8], mean time from repair 21.1 years [8.7]) who entered the study, 33 patients developed sustained monomorphic ventricular tachycardia, 16 died suddenly, and 29 had new-onset sustained atrial flutter or fibrillation. Electrocardiographic markers (QRS duration, QRS rate of change between 1985 and 1995) were significantly greater in the ventricular tachycardia and sudden-death groups. Older age at repair was associated with a higher risk of sudden death and atrial tachyarrhythmia. Pulmonary regurgitation was the main underlying haemodynamic lesion for patients with ventricular tachycardia and sudden death, whereas tricuspid regurgitation was for those with atrial flutter/fibrillation. Despite adverse haemodynamics, no patient who died suddenly had undergone late reoperation. CONCLUSION: Arrhythmia and sudden death are important late sequelae for patients after repair of tetralogy of Fallot. The electrophysiological and haemodynamic substrate of sudden death resembled that of sustained ventricular tachycardia, with pulmonary regurgitation being the predominant haemodynamic lesion. Preservation or restoration of pulmonary valve function may thus reduce the risk of sudden death. Furthermore, electrocardiographic markers can help to identify patients at risk. PMID- 11041399 TI - The role of acute and chronic stress in asthma attacks in children. AB - BACKGROUND: High levels of stress have been shown to predict the onset of asthma in children genetically at risk, and to correlate with higher asthma morbidity. Our study set out to examine whether stressful experiences actually provoke new exacerbations in children who already have asthma. METHODS: A group of child patients with verified chronic asthma were prospectively followed up for 18 months. We used continuous monitoring of asthma by the use of diaries and daily peak-flow values, accompanied by repeated interview assessments of life events and long-term psychosocial experiences. The key measures included asthma exacerbations, severely negative life events, and chronic stressors. FINDINGS: Severe events, both on their own and in conjunction with high chronic stress, significantly increased the risk of new asthma attacks. The effect of severe events without accompanying chronic stress involved a small delay; they had no effect within the first 2 weeks, but significantly increased the risk in the subsequent 4 weeks (odds ratio 1.71 [95% CI 1.04-2.82], p < or = 0.05 for weeks 2 4 and 2.17 [1.32-3.57], p < or = 0.01 for weeks 4-6). When severe events occurred against the backdrop of high chronic stress, the risk increased sharply and almost immediately within the first fortnight (2.98 [1.20-7.38], p < or = 0.05). The overall attack frequency was affected by several factors, some related to asthma and some to child characteristics. Female sex, higher baseline illness severity, three or more attacks within 6 months, autumn to winter season, and parental smoking were all related to increased risk of new exacerbations; social class and chronic stress were not. INTERPRETATION: Severely negative life events increase the risk of children's asthma attacks over the coming few weeks. This risk is magnified and brought forward in time if the child's life situation is also characterised by multiple chronic stressors. PMID- 11041400 TI - Frequency of choroidal abnormalities in neurofibromatosis type 1. AB - BACKGROUND: Choroidal neurofibromatosis is thought to be a rare form of neurofibromatosis that involves the eyes. The development of infrared light examination with a scanning laser ophthalmoscope (SLO) and indocyanine-green fundus angiography has allowed examination of the choroid. We studied choroidal abnormalities in patients with neurofibromatosis 1 and compared their frequency with that of other ocular abnormalities. METHODS: We examined 33 eyes of 17 consecutive patients diagnosed with neurofibromatosis 1 by conventional ophthalmoscopy and by non-invasive infrared monochromatic light with confocal SLO. 76 eyes of 39 age-matched controls were examined similarly by confocal SLO. 21 digital fluorescein and indocyanine-green angiographies were obtained from 11 adult patients, and 77 angiograms were obtained from age-matched controls. FINDINGS: Infrared monochromatic light examination by confocal SLO showed bright multiple patchy regions at and around the entire posterior pole of all 33 eyes examined. All bright patchy regions seen in adult patients corresponded to hypofluorescent areas on their indocyanine-green angiograms. However, no abnormalities were noted in any patient at corresponding areas under conventional ophthalmoscopic examination or fluorescein angiography. In SLO and indocyanine green studies, controls and control angiograms showed no choroidal abnormalities. Iris nodules were noted in 25 eyes (76%) of 14 patients (82%) and eyelid neurofibroma in five patients (29%). INTERPRETATION: The bright patchy regions noted under infrared fundus examination and the corresponding hypofluorescent areas seen on indocyanine-green angiograms are probably of choroidal origin. The high frequency (100%) of these abnormalities suggests that the choroid is one of the structures most commonly affected by neurofibromatosis 1. PMID- 11041401 TI - Impaired glucose tolerance and dyslipidaemia as late effects after bone-marrow transplantation in childhood. AB - BACKGROUND: This follow-up study aimed to assess the frequency of late effects on glucose and lipid metabolism after bone-marrow transplantation in childhood. METHODS: 23 long-term survivors (median age 20 years) were studied 3-18 years after bone-marrow transplantation and compared with 23 healthy controls matched for age and sex and with 13 patients in remission from leukaemia. FINDINGS: 12 (52%) of the 23 bone-marrow transplantation patients had insulin resistance, including impaired glucose tolerance in six and type 2 diabetes in four. The core signs of the metabolic syndrome (hyperinsulinaemia and hypertriglyceridaemia combined), were found in nine (39%) of the bone-marrow transplantation patients compared with one (8%) of the 13 leukaemia patients and none of the healthy controls (p=0.0015). The frequency of insulin resistance increased with the time since bone-marrow transplantation. Abdominal obesity, but not overweight, was common among the patients with insulin resistance. INTERPRETATION: Long-term survivors of bone-marrow transplantation are at substantial risk of insulin resistance, impaired glucose tolerance, and type 2 diabetes even at normal weight and young age. They also develop typical signs of the metabolic syndrome. We advocate measurement of serum lipids, fasting blood glucose, and serum insulin for the follow-up of all patients who undergo transplants in childhood, to be continued regularly and possibly life-long. PMID- 11041402 TI - Angina pectoris: a headache. PMID- 11041403 TI - Transmission of BSE by blood transfusion in sheep. AB - We have shown that it is possible to transmit bovine spongiform encephalopathy (BSE) to a sheep by transfusion with whole blood taken from another sheep during the symptom-free phase of an experimental BSE infection. BSE and variant Creutzfeldt-Jakob disease (vCJD) In human beings are caused by the same infectious agent, and the sheep-BSE experimental model has a similar pathogenesis to that of human vCJD. Although UK blood transfusions are leucodepleted--a possible protective measure against any risk from blood transmission--this report suggests that blood donated by symptom-free vCJD-infected human beings may represent a risk of spread of vCJD infection among the human population of the UK. PMID- 11041404 TI - Scleromyxoedema-like cutaneous diseases in renal-dialysis patients. AB - 15 renal dialysis patients have been identified with a skin condition characterised by thickening and hardening of the skin of the extremities and an increase in dermal fibroblast-like cells associated with collagen remodelling and mucin deposition. The disease closely resembles scleromyxoedema, yet has significant enough clinical and histopathological differences to warrant its designation as a new clinicopathological entity. PMID- 11041405 TI - Screening older adults at risk of falling with the Tinetti balance scale. AB - In a prospective study of 225 community dwelling people 75 years and older, we tested the validity of the Tinetti balance scale to predict individuals who will fall at least once during the following year. A score of 36 or less identified 7 of 10 fallers with 70% sensitivity and 52% specificity. With this cut-off score, 53% of the individuals were screened positive and presented a two-fold risk of falling. These characteristics support the use of this test to screen older people at risk of falling in order to include them in a preventive intervention. PMID- 11041406 TI - In-situ immuno-PCR to detect antigens. AB - In-situ immunoassays do not allow the detection of the minute numbers of target molecules accessible with in-situ PCR. We developed a highly sensitive method, termed in-situ immuno-PCR, in which the DNA marker was linked to target molecules through an antibody-biotin-avidin bridge and amplified by in-situ PCR. Amplified DNA sequences were detected in situ by hybridisation. This technique may be the only one available to detect minute quantities of biological macromolecules such as proteins, carbohydrates, and lipids in intact cells or tissue sections. PMID- 11041407 TI - Galactorrhoea and hyperprolactinaemia associated with protease-inhibitors. AB - We describe four patients with galactorrhoea as an isolated endocrine abnormality after use of protease inhibitors (PIs) as part of both highly active antiretroviral therapy (HAART) and postexposure prophylaxis (PEP). This reaction may be a direct effect of PIs or may be indirectly mediated by the effect of PIs on the cytochrome P450 system, thus potentiating the dopamine antagonist effects of other drugs. PMID- 11041408 TI - New data challenge OPV theory of AIDS origin. PMID- 11041409 TI - Past, present, and future of drug abuse at the Olympics. PMID- 11041410 TI - Japan's hibakusha still battle the effects of US nuclear bombs. PMID- 11041411 TI - US doctor pleads guilty to murdering patients. PMID- 11041412 TI - Researchers ask whether Dutch health care is inferior. PMID- 11041413 TI - Report welcomes slow but steady eradication of landmines worldwide. PMID- 11041414 TI - Intrauterine device and upper-genital-tract infection. AB - Concern about upper-genital-tract infection related to intrauterine devices (IUDs) limits their wider use. In this systematic review I summarise the evidence concerning IUD-associated infection and infertility. Choice of an inappropriate comparison group, overdiagnosis of salpingitis in IUD users, and inability to control for the confounding effects of sexual behaviour have exaggerated the apparent risk. Women with symptomless gonorrhoea or chlamydial infection having an IUD inserted have a higher risk of salpingitis than do uninfected women having an IUD inserted; however, the risk appears similar to that of infected women not having an IUD inserted. A cohort study of HIV-positive women using a copper IUD suggests that there is no significant increase in the risk of complications or viral shedding. Similarly, fair evidence indicates no important effect of IUD use on tubal infertility. Contemporary IUDs rival tubal sterilisation in efficacy and are much safer than previously thought. PMID- 11041415 TI - Fighting for public health along the USA-Mexico border. PMID- 11041416 TI - Takayasu's arteritis. PMID- 11041417 TI - Angiogenesis in chronic myelogenous leukaemia. PMID- 11041418 TI - 23-valent pneumococcal vaccination and HIV. PMID- 11041419 TI - 23-valent pneumococcal vaccination and HIV. PMID- 11041420 TI - Factor V Leiden paradox. PMID- 11041421 TI - Pluripotential cells in the bone marrow. PMID- 11041422 TI - Detection of hepatocellular damage. PMID- 11041423 TI - Directly observed therapy and treatment adherence. PMID- 11041424 TI - Directly observed therapy and treatment adherence. PMID- 11041425 TI - Directly observed therapy and treatment adherence. PMID- 11041426 TI - Directly observed therapy and treatment adherence. PMID- 11041427 TI - Neuroprotection failure in stroke. PMID- 11041428 TI - Consequences of breast screening. PMID- 11041429 TI - Evolution, immune response, and cancer. PMID- 11041430 TI - Evolution, immune response, and cancer. PMID- 11041431 TI - North and South: bridging the information gap. PMID- 11041432 TI - North and South: bridging the information gap. PMID- 11041433 TI - North and South: bridging the information gap. PMID- 11041434 TI - North and South: bridging the information gap. PMID- 11041435 TI - North and South: bridging the information gap. PMID- 11041436 TI - Ethics of identification. PMID- 11041437 TI - "Insiders" and "outsiders" in research collaborations? PMID- 11041438 TI - The precarious antioxidant defenses of the preterm infant. AB - Oxygen radicals are considered to be major causative factors in many illnesses of preterm infants. This article reviews the antioxidant defenses in immature animals and preterm infants, and attempts to quantitate their vulnerabilities to oxidants. Sources of oxidants, including hyperoxia, iron, parenteral nutrition, nitric oxide, and prooxidants, and their impact on immature antioxidant defenses are discussed. Genetic manipulations of antioxidant enzymes such as knockout and transgenic mice models are reviewed. The various clinical and investigational antioxidant therapies in animals and humans and difficulties in the design of antioxidant therapy studies are explored. PMID- 11041439 TI - Labor and delivery following successful external cephalic version. AB - The objective of this study is to determine if successful external cephalic version is followed by an increased likelihood of prolonged labor or operative delivery. Women having a successful external cephalic version of a normal singleton fetus > or =37 weeks' gestation between January 1, 1997 and December 31, 1998 were included. Each case was matched for gestational age at delivery (+/ 1 week), labor onset (spontaneous or induced), prior vaginal delivery (yes or no), and cervical dilation on admission for delivery (+/-1 cm) to the next three patients delivering a spontaneously vertex term singleton. Maternal demographics, intrapartum variables, neonatal outcomes, and route of delivery were examined. Statistical comparisons were performed by the Student's t-test or Fisher's exact test. The 38 cases and 114 controls were similar by maternal age, race, gestational age at delivery, birth weight, and insurer. There were no differences in the frequency of epidural or oxytocin use, maternal genital tract lacerations, or blood loss at delivery. Neonatal outcomes, assessed by 1- and 5-min Apgar score <7, or neonatal intensive care unit (NICU) admission did not differ between cases and controls. The labor length of patients undergoing successful version was similar to that of women laboring with spontaneously vertex fetuses (10.8 +/- 8.9 vs. 10.1 +/- 10.1 hr, p = 0.4). The frequencies of operative vaginal and cesarean delivery in cases did not differ from those of controls (3/38 vs. 1/114, p = 0.56 and 4/38 vs. 8/114, p = 0.51, respectively.) Labor duration and delivery route following successful external cephalic version do not differ from women with spontaneously vertex fetuses. PMID- 11041440 TI - Neonatal outcome in growth-restricted versus appropriately grown preterm infants. AB - The objective of this paper is to examine whether growth-restricted preterm infants have a different neonatal outcome than appropriately grown preterm infants. All consecutive, singleton preterm deliveries between 27-35 weeks' gestation were included over a 4-year period. Infants with congenital anomalies and infants of diabetic mothers were excluded. Infants were categorized as small for-gestational-age (SGA) when birth weight was at or below the 10th percentile, and appropriate-for-gestational-age (AGA) when between the 11th and 90th percentiles. Outcome variables included: neonatal death, respiratory distress syndrome (RDS), sepsis, intraventricular hemorrhage (IVH), and necrotizing enterocolitis (NEC). Neonatal morbidity and mortality were examined by univariate and stepwise multivariate logistic regression analyses. Factors controlled for during the analysis included: maternal age; gestational age; mode of delivery; presence of preeclampsia, HELLP syndrome, prolonged premature rupture of membranes (PROM), placental abruption, placenta previa, prenatal steroid exposure, infant gender, and low Apgar score. Seventy-six infants were included in the SGA group and 209 in the AGA group. SGA infants had a higher mortality rate (p = 0.003). They also had more culture-proven sepsis episodes (p = 0.001). No differences were found with respect to the other outcomes. The results were similar when analyzed separately for the group of infants born at or below 32 weeks' gestation. Growth-restricted preterm infants were found to have both higher mortality and infection rates compared with AGA preterm infants. Growth restriction in the preterm neonate was not found to protect against other neonatal outcomes associated with prematurity. When considering elective preterm delivery for this high-risk group of pregnancies, the increased risks in the neonatal period should be taken into account. PMID- 11041441 TI - Vitamin B12 and folate bioavailability from two prenatal multivitamin/multimineral supplements. AB - In this crossover, single-blind study, the bioavailability of B12 and folate, fasting and postprandially, was measured in 30 pregnant women for two prenatal multivitamin/multimineral supplements (Stuartnatal Plus and Materna, Wyeth-Ayerst Pharmaceuticals, Philadelphia, PA) and a placebo. Blood samples were obtained before supplementation and at 1, 3, 6, and 8 hr after supplementation serum levels of the two vitamins were measured by radioimmunoassay. The maximum postabsorption serum level was multiplied by the total body plasma levels to obtain the total rate of body absorption. The absorption peak of both vitamins occurred at 3 hours after ingestion of a supplement. The total body absorption of the two vitamins was greater during fasting than it was postprandially. There was 30% greater B12 absorption for Stuartnatal Plus (371 +/- 56 vs. 285 +/- 34 pmol) and 33% for Materna (315 +/- 34 vs. 236 +/- 4 pmol, p < or = 0.05). Similarly, there was 117% greater folate absorption fasting for Stuartnatal Plus (163 +/- 15 vs. 75 +/- 15 nmol, p < or = 0.001) and 57% greater absorption for Materna (207 +/- 21 vs. 132 +/- 13 nmol, p < or = 0.01). Both vitamins were readily absorbed (within 3 hours) into the maternal hepatic portal circulation. The absorption of both vitamins was significantly less when ingested after the test meal than when fasting. PMID- 11041442 TI - Perinatal lethal form of Gaucher's disease presenting with hemosiderosis. AB - A term infant with hydrops fetalis presented with hypotonia, massive splenomegaly, renal failure, and severe hyperferritinemia. Multiple organ failure, myoclonus, and opisthotonus ensued and she died at 15 days of age. High rounded forehead, large open fontanel, and a small recessed chin led to initial premortem diagnosis of Zellweger syndrome, but her plasma profile of long chain fatty acid was normal. Her subsequent clinical course and findings of postmortem examinations were consistent with perinatal lethal form of Gaucher's disease (PLGD). The diagnosis was confirmed by deficiency of enzyme beta glucocerebrosidase in white blood cells and in cultured fibroblasts. In addition to the crossover features of Zellweger phenotype, this infant exhibited a number of unusual features including, severe hyperferritinemia, rapid progression of splenomegaly, and absence of icthyosis. PMID- 11041443 TI - Hyperemesis gravidarum: a current review. AB - Hyperemesis gravidarum or pernicious vomiting of pregnancy affects between 0.3% and 2% of all pregnant patients. The objective of this paper is to review current literature pertaining to epidemiology, etiology, symptomatology, complications, treatment, and perinatal outcome of patients with hyperemesis gravidarum. We performed a MEDLINE search of the English literature from 1966 through January 2000 utilizing the keywords: hyperemesis gravidarum, nausea and vomiting, and pregnancy. Current data pertaining to epidemiology, etiology, clinical manifestations, differential diagnosis, complications, various treatment modalities, subsequent perinatal outcome and recent developments are presented. Review of the literature supports that hyperemesis gravidarum is a multifactorial disease in which pregnancy-induced hormonal changes associated with concurrent gastrointestinal dysmotility and possible Helicobacter pylori infection function as contributing factors. Therapeutic key elements are mainly supportive in conjunction with antiemetic medication. It appears perinatal outcome is unaffected. PMID- 11041444 TI - Recurrent familial neonatal deaths: hereditary surfactant protein B deficiency. AB - Hereditary surfactant protein B (SP-B) deficiency is an uncommon autosomal recessive lung disorder that causes hypoxemic respiratory failure in mature, morphologically normal infants. Recognition and diagnosis of this condition is of paramount importance, as it has significant implications for future pregnancies with a recurrence risk of 25%. In a family with three neonatal deaths over 20 years, SP-B deficiency was diagnosed following the death of the fourth affected infant. Previous deaths were mistakenly attributed to hyaline membrane disease (HMD), congenital Mycoplasma hominis infection, and pulmonary hypertension, however, following the diagnosis in the proposita, SP-B deficiency was also confirmed in her deceased siblings by immunohistochemical staining of autopsy specimens. This case highlights the presentation, postnatal course, diagnosis, and therapeutic options of SP-B deficiency in addition to the mode of inheritance and the possibility of antenatal diagnosis. Genetic consultation is imperative in the investigations of recurrent neonatal deaths, especially in cases of remote events. The recent enormous advances in human genetics have shown that many conditions previously ascribed to environmental agents have a genetic basis. PMID- 11041445 TI - Neuroendocrinology of the skin. AB - The classical observations of the skin as a target for melanotropins have been complemented by the discovery of their actual production at the local level. In fact, all of the elements controlling the activity of the hypothalamus-pituitary adrenal axis are expressed in the skin including CRH, urocortin, and POMC, with its products ACTH, alpha-MSH, and beta-endorphin. Demonstration of the corresponding receptors in the same cells suggests para- or autocrine mechanisms of action. These findings, together with the demonstration of cutaneous production of numerous other hormones including vitamin D3, PTH-related protein (PTHrP), catecholamines, and acetylcholine that share regulation by environmental stressors such as UV light, underlie a role for these agents in the skin response to stress. The endocrine mediators with their receptors are organized into dermal and epidermal units that allow precise control of their activity in a field restricted manner. The skin neuroendocrine system communicates with itself and with the systemic level through humoral and neural pathways to induce vascular, immune, or pigmentary changes, to directly buffer noxious agents or neutralize the elicited local reactions. Therefore, we suggest that the skin neuroendocrine system acts by preserving and maintaining the skin structural and functional integrity and, by inference, systemic homeostasis. PMID- 11041446 TI - Intercellular communication in the anterior pituitary. AB - In addition to hypothalamic and feedback inputs, the secretory cells of the anterior pituitary are influenced by the activity of factors secreted within the gland. The list of putative intrapituitary factors has been expanding steadily over the past decade, although until recently much of the work was limited to descriptions of potential interactions. This took the form of evidence of production within the pituitary of factors already known to influence activity of secretory cells, or further descriptions of actions on pituitary cells by such factors when added exogenously. A new phase of discovery has been entered, with extensive efforts being made to delineate the control of the synthesis and secretion of the pituitary factors within the gland, regulation of the receptors and response mechanisms for the factors in pituitary cells, and measurements of the endogenous actions of the factors through the use of specific immunoneutralization, receptor blockade, tissue from transgenic animals, and other means. Taken together, these findings are producing blueprints of the intrapituitary interactions that influence each of the individual types of secretory cells, leading toward an understanding of the physiological significance of the interactions. The purpose of this article is to review the recent literature on many of the factors acting as intrapituitary signals and to present such finding in the context of the physiology of the secretory cells. PMID- 11041447 TI - Endocrine and paracrine regulation of birth at term and preterm. AB - We have examined factors concerned with the maintenance of uterine quiescence during pregnancy and the onset of uterine activity at term in an animal model, the sheep, and in primate species. We suggest that in both species the fetus exerts a critical role in the processes leading to birth, and that activation of the fetal hypothalamic-pituitary-adrenal axis is a central mechanism by which the fetal influence on gestation length is exerted. Increased cortisol output from the fetal adrenal gland is a common characteristic across animal species. In primates, there is, in addition, increased output of estrogen precursor from the adrenal in late gestation. The end result, however, in primates and in sheep is similar: an increase in estrogen production from the placenta and intrauterine tissues. We have revised the pathway by which endocrine events associated with parturition in the sheep come about and suggest that fetal cortisol directly affects placental PGHS expression. In human pregnancy we suggest that cortisol increases PGHS expression, activity, and PG output in human fetal membranes in a similar manner. Simultaneously, cortisol contributes to decreases in PG metabolism and to a feed-forward loop involving elevation of CRH production from intrauterine tissues. In human pregnancy, there is no systemic withdrawal of progesterone in late gestation. We have argued that high circulating progesterone concentrations are required to effect regionalization of uterine activity, with predominantly relaxation in the lower uterine segment, allowing contractions in the fundal region to precipitate delivery. This new information, arising from basic and clinical studies, should further the development of new methods of diagnosing the patient at risk of preterm labor, and the use of scientifically based strategies specifically for the management of this condition, which will improve the health of the newborn. PMID- 11041448 TI - Mutations of gonadotropins and gonadotropin receptors: elucidating the physiology and pathophysiology of pituitary-gonadal function. AB - The recent unraveling of structures of genes for the gonadotropin subunits and gonadotropin receptors has provided reproductive endocrinologists with new tools to study normal and pathological functions of the hypothalamic-pituitary-gonadal axis. Rare inactivating mutations that produce distinctive phenotypes of isolated LH or FSH deficiency have been discovered in gonadotropin subunit genes. In addition, there is a common polymorphism in the LHbeta subunit gene with possible clinical significance as a contributing factor to pathologies of LH-dependent gonadal functions. Both activating and inactivating mutations have been detected in the gonadotropin receptor genes, a larger number in the LH receptor gene, but so far only a few in the gene for the FSH receptor. These mutations corroborate and extend our knowledge of clinical consequences of gonadotropin resistance and inappropriate gonadotropin action. The information obtained from human mutations has been complemented by animal models with disrupted or inappropriately activated gonadotropin ligand or receptor genes. These clinical and experimental genetic disease models form a powerful tool for exploring the physiology and pathophysiology of gonadotropin function and provide an excellent example of the power of molecular biological approaches in the study of pathogenesis of diseases. PMID- 11041449 TI - Ceramide-induced apoptosis of human thyroid cancer cells resistant to apoptosis by irradiation. AB - Ionizing radiation (IR) induces apoptosis through, in part, cell membrane breakdown signals. Ceramide and diacylglycerol (DAG) are released after IR exposure, which act as second messengers to induce proapoptotic and antiapoptotic signals, respectively. We have previously shown, however, that thyroid cells are relatively resistant to IR-induced apoptosis. To investigate the mechanism of thyroid cell resistance to IR-related apoptosis, we determined the effects of ceramide and its release following exposure of human thyroid cancer cell lines to IR. Exogenous C2-ceramide (10-100 microM) activated the apoptosis process in all cell lines used. Exogenous C2-ceramide also activated a stress kinase, c-Jun N terminal kinase UNK). The apoptotic action of ceramide was attenuated by serum or simultaneous activation of protein kinases C and A by phorbol esters and forskolin. Furthermore, 2-5 Gy IR had a differential effect on ceramide and DAG release in human thyroid cells; a weak and transient release of ceramide but a strong and sustained release of DAG. Our results indicated that the radioresistance properties of thyroid cancer cells probably reflect the dominance of anti-apoptotic signals, evoked by growth factor(s) and DAG, which override the apoptotic effect of ceramide released by human thyroid cells on exposure to IR, in spite of activation of proapoptotic pathway downstream of ceramide. PMID- 11041450 TI - Cyclin D1 overexpression in thyroid carcinomas: relation with clinico pathological parameters, retinoblastoma gene product, and Ki67 labeling index. AB - Cyclin D1 is a G1 cyclin participating in the control of cell cycle progression through interaction with the retinoblastoma gene product (pRB). The overexpression of positive regulators (such as cyclin D1) has been reported in a variety of neoplasms, but their role in thyroid tumorigenesis is yet to be established. In our series of 54 thyroid carcinomas, cyclin D1 overexpression (detected by both immunohistochemistry and by Northern blotting) was correlated with prognostic variables, proliferative activity and pRB. Cyclin D1 overexpression was observed in 35% of thyroid carcinomas with a significantly higher expression of this cyclin in neoplastic tissues than in matched normal parenchyma. In well-differentiated carcinomas, the cyclin D1 mRNA overexpression was inversely correlated with nodal status (p = 0.03), while the protein product was higher in tumors from patients less than 40 than patients over 40 years of age. Inversely, there was no significant correlation with gender and tumor status, pRB and with proliferative activity. PMID- 11041451 TI - The thyrotropin receptor is not involved in the activation of p42/p44 mitogen activated protein kinases by thyrotropin preparations in Chinese hamster ovary cells expressing the human thyrotropin receptor. AB - We studied whether bovine pituitary thyrotropin (bTSH) or human recombinant thyrotropin (rhTSH) stimulated p42/p44 mitogen-activated protein kinases (MAPKs) in Chinese hamster ovary cells expressing human thyrotropin receptor (CHO-hTSHR cells). We show that p42/p44 MAPK phosphorylation was induced by both TSH preparations at similar levels in CHO-hTSHR cells and in wild-type CHO cells. In contrast, cyclic adenosine monophosphate (cAMP) production was stimulated by TSH only in CHO-hTSHR cells, demonstrating that p42/p44 MAPK stimulation was independent of the TSH receptor. Moreover, similar results were obtained with two other cell lines: the FRTL-5 thyroid cell line and the CCL39 fibroblast cell line. Maximal stimulation of p42/p44 MAPK phosphorylation was observed after a 5- to 10-minute incubation with bTSH and rhTSH preparations. At this time, the phosphorylation of GST-Elk1 was also increased in a time- and concentration dependent manner by bTSH preparations. The phosphorylation of p42/p44 MAPKs was abolished by PD 98059 and GF 109203X, indicating the involvement of MAPK kinases (MEK 1/2) and protein kinase C. In contrast, the activation of p42/p44 MAPKs was insensitive to H89, to cholera toxin and to pertussis toxin. These data suggest that the protein kinase A pathway was not implicated in p42/p44 MAPK activation by TSH preparations. Moreover, Gs or Gi/Go proteins do not appear to participate in p42/p44 MAPK activation. We also showed that these TSH preparations failed to induce activation of c-Jun NH2 terminal kinase. We therefore conclude that the commercial TSH preparations used in this study contained factor(s) responsible for the specific activation of p42/p44 MAPKs by a TSH receptor-independent mechanism. PMID- 11041452 TI - Thyroid hormone stimulates Na, K-ATPase gene expression in the hemodynamically unloaded heterotopically transplanted rat heart. AB - Regulation of myocardial Na, K-ATPase gene expression by thyroid hormone was investigated in the heterotopically transplanted rat heart to distinguish the direct effects of the hormone on the heart from effects secondary to increased hemodynamic workload. In this model, the transplanted heart is histologically normal and spontaneously beating, but hemodynamically unloaded. Three days after transplantation, relative contents of ventricular Na, K-ATPase alpha2- and beta1 mRNAs and alpha1- and alpha2-proteins were increased twofold to threefold in the transplanted heart, but these changes were transient. We next determined the maximal triiodothyronine (T3)-induced changes that are observed in various parameters of Na, K-ATPase expression in the heart: treatment of nontransplanted euthyroid rats with T3 to reach hyperthyroid steady state resulted in significant increases in heart weight, RNA and RNA/protein ratio, Na, K-ATPase activity, Na, K-ATPase alpha2-protein and enzyme activity, and approximately threefold increase in both alpha2- and beta1-mRNA content. The effect of treatment with thyroxine (T4) on the heterotopically transplanted and the in situ heart was then examined. T4 treatment (of the host) resulted in a significant increase in Na, K-ATPase alpha1-, alpha2-, and beta1-mRNAs in transplanted hearts (1.6 +/- 0.1-, 2.4 +/- 0.2-, and 1.7 +/- 0.1-fold, respectively), that was associated with a 2.2 +/- 0.2 fold increase in alpha2 protein as compared to transplanted hearts in diluent treated euthyroid hosts (p < 0.05 for all changes). In addition, T4-induced increments in transplanted hearts were similar to those observed in the corresponding in situ hearts of host rats treated with T4. We conclude that the increase in Na, K-ATPase expression by thyroid hormone largely occurs independently of increased cardiac work elicited by the hormone and reflects a direct action of the hormone on Na, K-ATPase gene expression. PMID- 11041453 TI - No evidence of thyrotropin receptor and G(s alpha) gene mutation in high iodine uptake thyroid carcinoma. AB - Usually, thyroid carcinoma presents as a cold nodule on radioiodine scintigraphy. High-uptake nodules on iodine thyroid scans are associated with an exceedingly low incidence of malignancy. Only 29 cases of carcinomas appearing as hot or warm nodules have as yet been reported. From 1993 to 1999, we have observed eight similar cases (4 hot and 4 warm thyroid nodules) suggesting that thyroid carcinomas may not be as rare as usually considered in these circumstances. Four tumors were available for molecular analysis on paraffin-embedded sections. Because no mutations were found in the whole coding portions of thyrotropin receptor (TSH-R) gene and fragments encompassing the mutational hot spots of the G(s alpha) gene, it is unlikely that activating mutations of the TSH-R or G(s alpha) genes were involved in these carcinomas. PMID- 11041454 TI - Using recombinant human TSH in the management of well-differentiated thyroid cancer: current strategies and future directions. AB - Mortality rates from thyroid cancer have fallen significantly in recent decades, almost certainly as the result of earlier diagnosis and improved treatment of differentiated (papillary and follicular) thyroid cancer. Enhanced survival is likely a result of early diagnosis and therapy applied at a disease stage when treatment is most effective. In the United States and Europe, most patients at high risk for relapse and death from thyroid cancer are treated with total or near-total thyroidectomy and receive radioiodine ablation of residual normal or malignant thyroid tissue, followed by treatment with thyroid hormone, a strategy that cures more than 80% of patients. Still, some die of the disease and nearly 15% have local recurrences, while another 5% to 10% develop distant metastases. Over 50% of recurrences appear in the first five years, but distant metastases may surface years, and sometimes decades, after initial therapy. Much has been learned about risk stratification to predict recurrence and death from thyroid cancer but individual patients continue to have adverse outcomes not always foreseen by a low tumor stage. Follow-up must accordingly be meticulous and prolonged. The National Cancer Center Network (NCCN) has recently established consensus practice guidelines that give explicit advice about the diagnosis and management of benign and malignant thyroid tumors, including paradigms for long term follow-up and the treatment of recurrent disease. The guidelines confirm that diagnostic scanning with 131I and measurement of serum thyroglobulin (Tg) levels are the mainstay of follow-up, offering the opportunity to detect recurrent or persistent cancer at very early stages. These guidelines advocate TSH-stimulated serum Tg measurements, done either during thyroid hormone withdrawal or stimulation with recombinant human TSH (rhTSH, Thyrogen), that often identify the presence of cancer well before diagnostic whole-body scanning or other imaging studies can spot the tumor, which offers the opportunity to treat recurrent disease at an early stage. The use of rhTSH adds a new dimension to long-term follow-up that avoids putting patients through the symptoms of hypothyroidism, and offers the opportunity to follow some patients with rhTSH stimulated serum Tg levels without performing 131I whole-body scans. A multicenter international study has shown that serum Tg measurements alone are not as sensitive in the identification of patients with persistent or recurrent tumor as are rhTSH-stimulated serum Tg determinations. Although not yet approved for preparation of patients for 131I therapy, rhTSH has been used successfully in a compassionate use program for this purpose in a relatively large number of patients. Formal clinical investigations now planned to provide guidelines for the use of rhTSH for therapeutic 131I portend a new set of effective therapeutic paradigms for the management of differentiated thyroid cancer. PMID- 11041455 TI - Accuracy considerations when using early (four- or six-hour) radioactive iodine uptake to predict twenty-four-hour values for radioactive iodine dosage in the treatment of Graves' disease. AB - Although literature has offered methods to predict 24-hour radioactive iodine uptake values from early (4- to 6-hour) measurements, the resultant dosage errors have not been examined. Potential errors include underdosage, overdosage, and a failure to recognize rapid turnover patients (early-to-late uptake ratios > or = 1) who are at high risk for treatment failure and full-body radiation exposure. We developed and tested a novel method for minimizing error involved in using a single early uptake measurement to derive late uptake. From a retrospective analysis of 203 Graves' disease patients, receiver operating characteristic (ROC) curve analysis enabled us to identify patients likely to experience rapid turnover and therefore should receive 24-hour studies. Twenty-four-hour uptake measurements are necessary with 77% or more 4-hour uptake values and 80% or more 6-hour values. After eliminating these patients, we developed linear regression equations to predict the 24-hour uptake from 4-hour (n = 61) and 6-hour (n = 22) rule groups, testing their efficacy on separate 4-hour (n = 61) and 6-hour (n = 21) patient groups. We also used our test population to measure error in four early-to-late uptake conversion formulas presented in the literature. Error involved in these predictions ranged from a 10.6% overestimate for 4-hour calculations to a 5.9% underestimate for 6-hour calculations. When applied to two dosage formulas incorporating gland size, absorbed dose, and 24-hour uptake, average dosage error was 7%. In comparison to the other sources of error radioactive iodine (131I) dosimetry, potential error in predicting 24-hour uptake from 4- or 6-hour uptake values is low. PMID- 11041456 TI - Limited genetic susceptibility to severe Graves' ophthalmopathy: no role for CTLA 4 but evidence for an environmental etiology. AB - Graves' disease (GD) is an autoimmune thyroid disease (AITD) characterized by hyperthyroidism and by the occurrence of a distinctive ophthalmopathy (orbitopathy), which presents with varying degrees of severity. Graves' disease clusters in families but the importance of heredity in the pathogenesis of the associated ophthalmopathy is unclear. We have studied the family history of 114 consecutive, ethnically mixed patients with severe Graves' ophthalmopathy (GO). Patients were selected by unambiguous single ascertainment. Seventy-seven percent of patients were female and 59% smoked. The mean age at onset of their GD was 43 years (range 17-78 years). Forty-one patients (36%) had a family history of AITD, defined as a first- and/or a second-degree relative affected with either Graves' disease (GD) or Hashimoto's thyroiditis (HT). The segregation ratio for AITD in nuclear families in our ascertained Graves' ophthalmopathy families was 0.07 (0.12 in Caucasians only). Hence, the higher prevalence of AITD among relatives of Graves' ophthalmopathy patients agreed with the known genetic predisposition to AITD and this predisposition was stronger in women than in men. However, only 3 of the 114 patients had a family history of severe Graves' ophthalmopathy (all second-degree relatives) and the segregation ratio for GO was 0. These data did not support a major role for familial factors in the development of severe Graves' ophthalmopathy distinct from Graves' disease itself. In addition, we tested 4 candidate genes, human leukocyte antigen (HLA), tumor necrosis factor beta (TNF-beta), CTLA-4 and the thyrotropin receptor (TSHR), for association with Graves' ophthalmopathy. These were negative except for the HLA and CTLA-4 genes, which were found to be weakly associated with GO giving similar relative risk (RR) as in GD patients without ophthalmopathy. These data suggested that environmental factors, rather than major genes, were likely to predispose certain individuals with AITD to severe Graves' ophthalmopathy. Smoking remains one example of such potential external insults. PMID- 11041457 TI - Increased frequency of euthyroid ophthalmopathy in patients with Graves' disease associated with myasthenia gravis. AB - We previously showed that myasthenia gravis (MG) has a mild clinical expression when associated with autoimmune thyroid diseases (AITD). In the present study we have investigated the frequency of thyroid-associated ophthalmopathy (TAO) in patients with Graves' disease (GD) associated with MG as compared with GD patients without MG. A total of 418 patients with GD were studied, 31 with MG and 387 without MG. TAO was evaluated by physical examination, exophthalmometry, computerized tomography, and computerized visual fields assessment. The overall prevalence of TAO was similar in GD patients with MG (61.2%) and in those without MG (56.4%). When the analysis was restricted to GD patients with ocular MG, a greater frequency of TAO was found (84.6%), compared with GD patients without MG or with GD patients with generalized MG, although the differences did not reach the statistical significance. GD patients with MG had a significantly greater prevalence (12.9%) of euthyroid ophthalmopathy (clinically overt ophthalmopathy without previous and/or current hyperthyroidism) than those without MG (3.1%; p = 0.003). The results suggest a preferential association between the ocular manifestations of GD and MG, which may be due to immunological cross-reactivity against common autoimmune targets in the eye muscle as well as to a common genetic background. PMID- 11041458 TI - Changes in lipoprotein(a) levels in overt and subclinical hypothyroidism before and during treatment. AB - The aim of this prospective, follow-up study was to examine the influence of overt hypothyroidism (OHP) and subclinical (SHP), before and during thyroxine (T4) treatment, on lipoprotein(a) [Lp(a)], other lipoproteins, and apolipoproteins. Twenty-four patients (17 females, 7 males) with OHP, aged 54 +/- 11.1 years (group A) and 23 patients (females) with SHP aged 50.1 +/- 13.2 years (group B) were evaluated and compared to 34 and 38 controls, respectively. All patients received T4 therapy in a stepwise fashion until euthyroidism was reached. Thyrotropin (TSH), free thyroxine (FT4), and total triiodothyronine (TT3) levels were measured before T4 therapy and repeatedly every 4 weeks after the initiation of treatment until the euthyroid state was reached. Levels of Lp(a), total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDLc), high-density lipoprotein cholesterol (HDLc), apolipoprotein A1 (apoA1) and apolipoprotein B (apoB) were measured before and 4 months after the achievement of euthyroidism. Additionally, body mass index (BMI) was also evaluated. We found that in OHP patients, levels of TC, LDLc, and apoB were elevated before treatment and decreased significantly after the return to the euthyroid state. BMI and levels of triglycerides also decreased significantly; Lp(a) was higher in OHP patients in comparison with controls and decreased significantly by 14.56% (25.29% in men and 10.34% in women) during T4 treatment. In SHP patients, levels of all common lipoproteins, apolipoproteins, and Lp(a) did not differ significantly from controls before treatment and did not change after the euthyroid stage was reached. It is concluded that in overt hypothyroidism, Lp(a) levels and most of the lipoproteins were elevated before treatment and decreased significantly. In subclinical hypothyroidism, lipoproteins and Lp(a) levels were normal at baseline and did not change during treatment. PMID- 11041459 TI - Thyroid-stimulating antibody is related to Graves' ophthalmopathy, but thyrotropin-binding inhibitor immunoglobulin is related to hyperthyroidism in patients with Graves' disease. AB - We investigated the relationship between thyroid function or ophthalmopathy of Graves' disease and thyrotropin receptor antibodies (TRAb) in 155 untreated patients with Graves' hyperthyroidism. All patients were examined by ophthalmologists, and underwent computed tomography of the orbit and measurement of serum free triiodothyronine (FT3), free thyroxine (FT4), thyrotropin-binding inhibitor immunoglobulin (TBII), and thyroid stimulating antibodies (TSAb). Patients were divided into three groups according to the presence of orbital fat increase (OFI) and extraocular muscle enlargement (EME): 57 patients without OFI and EMO formed the no Graves' ophthalmopathy (NGO) group; 55 patients with OFI but without EMO formed the OF group; 43 patients with EME with or without OFI formed the EM group. The FT3, FT4, and thyroid weight increased in the order of the EME, NGO, and OFI groups. TSAb increased in the order of the NGO, OFI, and EME groups, and TSAb was significantly greater in the EME and OFI than in the NGO group. TBII was not significantly different among the three groups, but was lower in EME than NGO. There was a significant positive correlation between TBII and FT3 or FT4 in all patients combined as well as in all three groups, but correlation between TSAb and FT3 or FT4 was very weak in all groups, and that between TSAb and FT3 was not significant in the EM group In the relationship between ophthalmopathy and TRAb, the sum of the scores of eyelid swelling, proptosis, and extraocular muscle enlargement was taken as a measure of the overall severity of the Graves' ophthalmopathy (GO). TSAb was significantly correlated with the GO score, but there was no correlation between TBII and GO scores. In conclusion, TSAb was correlated with ophthalmopathy but TBII was related to hyperthyroidism. PMID- 11041460 TI - Increased levels of serum interleukin-18 in Graves' disease. AB - We have previously reported that serum soluble interleukin-2 receptor (sIL-2R) and IL-12 levels were significantly increased in hyperthyroid Graves' disease. In this study, we investigated serum IL-18 levels in patients with either Graves' disease or Hashimoto's thyroiditis. The serum IL-18 levels in Graves' disease were significantly increased in the hyperthyroid state and were decreased during treatment with methimazole or propylthiouracil. On the other hand, the levels in Hashimoto's thyroiditis in the hypothyroid state showed no significant differences from those in healthy subjects. When liothyronine sodium was administered orally to healthy subjects, serum IL-18 levels were not changed. Positive correlations between serum IL-18 and IL-12, IL-12 and sIL-2R, and sIL-2R and IL-18 levels were noted in Graves' disease. These results suggest that Th1 cytokines might play an important regulatory role in Graves' disease. PMID- 11041461 TI - Hypothyroidism and aging: the Rosses' survey. AB - An earlier impression of a high prevalence of hypothyroidism in a general practice (4,190 patients including 1,544 adult females aged 18 years or more with 544 aged 50 years or more) in the Rosses, a coastal area in the northwest of Ireland was confirmed by this study. The accumulated prevalence of overt spontaneous primary hypothyroidism was 8.6% in 544 females aged 50 years or more but only 0.9% in the 1,000 females between 18 and 50 years of age. This prevalence was approximately twice that of an Irish National general practice population sample of 4,314 females aged 50 years or more (8.6% vs. 4.6%) p < 0.001. The reasons for this difference are unclear but may reflect the high level of opportunistic screening carried out in West Donegal. Thyroid peroxidase antibodies measured by radioimmunoassay were found in 75.6% of hypothyroid patients compared to 18.6% of practice controls (p < 0.01). Neither HLA-DRB1, DQA1, and DQB1 phenotype frequencies nor dietary iodine intake (median urinary iodine excretion 104 microg/L) appeared to be contributory factors. The finding of an 8.6% accumulated prevalence of hypothyroidism in females greater than 50 years of age when a population is aggressively investigated demonstrates the relative importance of its contribution to total morbidity and suggests that the disorder may be underdiagnosed, thus supporting the concept of targeted screening in this age group. PMID- 11041462 TI - Development of overt autoimmune hyperthyroidism in a patient therapeutically immunosuppressed after liver transplantation. AB - Immunosuppression is a therapeutic maneuver directed at preventing transplant rejection. When applied to autoimmunity, immunosuppression is intended to target similar immune processes. We report an unusual case of a 35-year-old woman who developed autoimmune hyperthyroidism of Graves' disease while on immunosuppressive therapy for liver transplantation. Signs and symptoms of hyperthyroidism were already present when, misled by the concomitant toxic hepatic syndrome, liver rejection was first suspected. Despite a therapeutic level of cyclosporine, elevated serum alanine and aspartate aminotransferase levels were noted. Consequently, a liver biopsy was performed to exclude an acute rejection. The findings were consistent with acute hepatitis without evidence of rejection. Then, the diagnosis of Graves' hyperthyroidism was considered and finally confirmed by finding a suppressed thyroid-stimulating hormone, elevated thyroid hormone levels, and a high and homogeneous thyroid uptake from radioactive iodine scan. Thyroid peroxidase antibody and thyroid-stimulating immunoglobulin were markedly elevated. The patient was treated with radioactive iodine, which resulted in improvement of symptoms and resolution of abnormal liver function tests. Although the mechanisms involved in transplant rejection and human autoimmunity are thought to be similar, the development of Graves' disease in this patient despite therapeutic immunosuppression suggests that the immunological processes may be different. PMID- 11041463 TI - Concomitant presentation of Hashimoto's thyroiditis and maltoma of the thyroid in a twenty-year-old man with a rapidly growing mass in the neck. AB - We report an uncommon case of a 20-year-old man, who noted a painless, growing mass in his neck, which appeared in a weekend, associated with moderate dysphagia and weakness. Laboratory examination revealed an elevated serum thyrotropin of 25 mU/L, normal serum triiodothyronine and thyroxine levels, and high titers of antithyroglobulin and antithyroid peroxidase antibodies. The neck lesion showed a depressed iodine uptake in the left thyroid lobe, which had an asymmetrical pseudocystic pattern associated with poor vascularization in the ultrasound scan. Cytologic examination showed a lymphocyte thyroiditis in association with lymphoma of large cell arising from mucosa-associated lymphoid tissue (MALT lymphoma or maltoma). The patient underwent a left thyroid lobectomy while being treated with levothyroxine for Hashimoto's thyroiditis, and the surgical treatment was further complemented with chemotherapy using fludarabine. The histologic examination confirmed the cytologic findings and the immunohistochemistry showed a B-cell type maltoma. Additional investigation provided no evidence of disease in other tissues. The clinical course has been favorable in the first 2 years of follow-up, with no evidence of local or systemic recurrence of the disease. PMID- 11041464 TI - Images in thyroidology. Left exophthalmos of endocrine nonthyroid origin in a seven-year-old boy. PMID- 11041465 TI - Noninvasive vascular laboratory for evaluation of peripheral arterial occlusive disease: Part I--hemodynamic principles and tools of the trade. PMID- 11041466 TI - Fibrin sheath stripping versus catheter exchange for the treatment of failed tunneled hemodialysis catheters: randomized clinical trial. AB - PURPOSE: To compare the effectiveness of two treatments for tunneled hemodialysis catheter malfunction: percutaneous fibrin sheath stripping (PFSS) and over-the wire catheter exchange (EX). MATERIALS AND METHODS: Adult patients with poorly functioning tunneled hemodialysis catheters (flow rates < 200 mL/min) were randomly assigned to receive either PFSS or EX. Over the course of 20 months, 30 patients (37 encounters) referred to a single institution met the inclusion criteria and consented to participate. PFSS employed transcatheter snares via femoral vein puncture, whereas EX was performed over a guide wire with use of fluoroscopic guidance. Patients were followed up to determine the duration of continued adequate hemodialysis via manipulated catheters for up to 4 months (primary outcome measure). RESULTS: Overall technical success rate was 97%. Mean catheter patency for the PFSS group was 24.5 +/- 29.3 days, and 52.2 +/- 43 days for the EX group (P < .0001). After EX, patency rates at 1, 2, 3, and 4 months were 71%, 33%, 27%, and 27%, compared to 31%, 16%, 7%, and 0% after PFSS (P = .04, logrank test). Exchanged catheters were significantly more likely to be patent for as long as 4 months (23% versus 0%; P < .05, chi2 test). CONCLUSIONS: Malfunctioning tunneled hemodialysis catheters treated by means of EX are significantly more likely to remain patent for up to 4 months than are those treated by means of PFSS. According to the results of this trial, PFSS should not be performed as a routine therapy for catheter malfunction. PMID- 11041467 TI - Percutaneous fibrin sheath stripping versus transcatheter urokinase infusion for malfunctioning well-positioned tunneled central venous dialysis catheters: a prospective, randomized trial. AB - PURPOSE: To compare central dialysis catheter patency rates after stripping procedures with those after urokinase (UK) infusion. MATERIALS AND METHODS: Fifty seven tunneled catheters with either (i) flow rates less than 250 mL/min and established baseline flow rates > or = 300 mL/min or (ii) flow rates 50 mL/min less than higher established baseline flows were prospectively randomized to undergo stripping procedures (n = 28) or UK infusion (n = 29) at 30,000 U/h via each port concurrently, for a total 250,000 U. Success and patency were determined by dialysis at normal flow rates (> or = 300 mL/min) or at the previously established higher baseline rate. Flow rates were monitored weekly. Primary patency ended with catheter malfunction or removal. Kaplan-Meier survival analysis was used to construct survival curves. RESULTS: In the stripping group, initial clinical success was 89% (25 of 28). The 15-, 30-, and 45-day primary patency rates were 75% (n = 20), 52% (n = 13), and 35% (n = 8), respectively. The median duration of additional function was 32 days (95% CI: 18-48 d). In the UK group, initial clinical success was 97% (28 of 29). The 15-, 30-, and 45-day primary patency rates were 86% (n = 21), 63% (n = 13), and 48% (n = 9), respectively. The median duration of additional patency was 42 days (95% CI: 22 153 d). The Wilcoxon test for equality detected no significant difference in the survival curves for the two treatment groups (P = .236). CONCLUSION: There is no significant difference in time to primary patency between the two methods. Both allow temporary catheter salvage in most patients. PMID- 11041469 TI - Conversion of indwelling chest port catheters to tunneled central venous catheters. AB - PURPOSE: To determine the safety and efficacy of the conversion of subcutaneous chest wall infusion ports to tunneled central venous catheters. MATERIALS AND METHODS: During a period of 34 months, 67 patients were referred for conversion of indwelling subcutaneous chest wall ports to tunneled central venous catheters as part of a bone marrow transplant protocol. Six patients were deemed unacceptable for conversion and the remaining 61 underwent successful conversion. All patients had functioning surgically placed single-lumen (n = 50) or double lumen (n = 11) chest ports, which were removed to maintain the original venous access sites for placement of a tunneled central venous catheter, incorporating the chest wall pocket for tunneling, in 46 patients (75%). A new tunnel was created in the other 15 patients. There were no immediate complications and all patients were followed until catheter removal or patient demise with the catheter in place. RESULTS: 57 of 61 (93%) catheters were used without evidence of infection for 23-164 days (mean, 57 d) after placement. Two (3%) were removed (both at 26 days) because of persistent neutropenic fever without physical signs or laboratory evidence of catheter infection, and two (3%) were removed (at 11 and 77 days) because of proven catheter infection, yielding an overall infection rate of 1.2 per 1,000 catheter days. Two catheters required exchange and two required stripping because of decreased function, resulting in an overall catheter-related complication rate of 2.4 per 1,000 catheter days. CONCLUSIONS: Indwelling subcutaneous chest wall infusion ports can be safely converted to tunneled central venous catheters, even in an immunocompromised patient population, with a low risk of complications such as infection. PMID- 11041468 TI - Hemodialysis catheter-associated fibrin sheaths: treatment with a low-dose rt-PA infusion. AB - PURPOSE: To prospectively evaluate the efficacy of a low-dose, 3-hour recombinant tissue plasminogen activator (rt-PA) infusion for the treatment of hemodialysis catheter (HDC)-associated fibrin sheaths. MATERIALS AND METHODS: Seventeen patients with end-stage renal disease (female, n = 11; male, n = 6), who were undergoing catheter-directed hemodialysis, were evaluated for 28 episodes of HDC dysfunction. This patient group ranged in age from 25 to 92 years (mean, 57 years). Radiographic contrast and/or clinical evaluation were consistent with the presence of a fibrin sheath on either the arterial and/or venous port in all cases. Patients subsequently underwent a thrombolytic infusion consisting of 2.5 mg rt-PA in 50 mL normal saline at a rate of 17 mL/h (3-hour infusion) per port. All infusions were performed in the interventional radiology recovery room, on an outpatient basis. Patients were followed-up prospectively for technical success, complications, catheter patency, and long-term outcome. RESULTS: The immediate technical success rate, defined as return of manual aspiration and infusion capabilities to both ports, was 100%. No potential patients required exclusion from thrombolytic therapy secondary to contraindications, and no procedure related complications occurred. The arithmetic mean and median catheter patency at the end of the study was 41 and 25 days, respectively (range, 1-116 days). A Kaplan-Meier survival analysis yielded a 30-, 60-, and 90-day probability of patency of 0.67, 0.61, and 0.51, respectively. At the end of the study period, all 17 patients remained on catheter-directed hemodialysis and 13 (76%) were utilizing the same catheter present at the time of entrance into the study. CONCLUSION: Thrombolytic therapy utilizing a 2.5-mg rt-PA infusion through each port during a 3-hour period would appear to be a safe, efficient method for treating HDC-associated fibrin sheaths. Three-month patency rates are comparable to those reported for other methods of restoring function to HDC catheters, including new catheter placement, catheter exchange over a guide wire, thrombolytic infusions with urokinase, and percutaneous fibrin sheath stripping. PMID- 11041470 TI - Pulmonary embolism from pulse-spray pharmacomechanical thrombolysis of clotted hemodialysis grafts: urokinase versus heparinized saline. AB - PURPOSE: To compare the frequency and extent of pulmonary embolism (PE) occurring during pulse-spray pharmacomechanical thrombolysis (PSPMT) of clotted hemodialysis grafts with use of either urokinase (UK) or heparinized saline (HS). Postintervention primary patency and complication rates were compared for each method of thrombolysis. METHODS AND MATERIALS: Twenty-seven patients were enrolled in this prospective, randomized, double-blind study evaluating PE with two PSPMT agents. The doses of heparin were similar between groups. The only variable was that one group of patients received UK and the other received HS. In two cases, the venous anastomosis could not be crossed. Eleven patients were treated with UK and 14 with HS. Nuclear medicine perfusion lung scans were performed before treatment and after graft declotting procedures. Lung perfusion was quantified to 10% of a pulmonary segment (0 = normal perfusion, 1 = segmental perfusion defect), with nine segments counted for each lung. RESULTS: Baseline nuclear medicine perfusion lung scan results were abnormal (> or = 20% segmental perfusion defect) in 19 patients (70.4%). New PE (one or more pulmonary segments) occurred in two patients treated with UK (18.2%) and nine patients treated with HS (64.3%; P = .04). All cases of PE were asymptomatic. Quantitative global pulmonary perfusion analyses revealed that treatment with UK improved flow to 0.2 +/- 2.0 pulmonary segments, whereas treatment with HS decreased perfusion to 1.0 +/- 1.7 segments (P = .16, NS). Although postintervention primary patency rates were similar according to life-table analysis (P = .76, NS), complication rates were higher with use of HS (n = 4, 28.6%) than with use of UK (n = 2, 18.2%) (P = .6, NS). CONCLUSIONS: All PE were asymptomatic during PSPMT, but treatment with UK reduced the rate of PE and tended to result in smaller defects in lung scan results. Most patients undergoing hemodialysis have abnormal baseline perfusion scan results, but PSPMT with UK improved many of them. The postintervention primary patency rates were similar between groups, but complications were more frequent after treatment with HS. PMID- 11041471 TI - Mechanical thrombolysis of thrombosed hemodialysis native fistulas with use of the Arrow-Trerotola percutaneous thrombolytic device: our preliminary experience. AB - PURPOSE: To evaluate the feasibility of use of the Arrow-Trerotola percutaneous thrombolytic device (PTD) in the treatment of thrombosed hemodialysis native fistula occlusions. MATERIALS AND METHODS: Ten patients with native fistula occlusion underwent mechanical thrombolysis with use of the PTD. The standard PTD was used in seven patients and the over-the-wire device was used in three patients. Major outcomes of our study included procedure time, clinical success, complication rate, and 3- and 6-month patency rates. RESULTS: The technical success rate was 100% and the clinical success rate was 90% (9 of 10). In all 10 cases, the procedure was associated with angioplasty. There were no major complications. The mean time of successful procedures was 126.1 minutes. The 3- and 6-month primary patency rates were 70% and 60%, respectively; the assisted primary patency rate at 6 months was 80%. CONCLUSION: The PTD is an effective mechanical device for percutaneous treatment of thrombosed hemodialysis access. Our clinically successful initial experience with the PTD shows that the technique is rapid and safe for treatment of native fistula occlusions. PMID- 11041472 TI - Massive pulmonary embolism: treatment with the hydrolyser thrombectomy catheter. AB - PURPOSE: To assess the efficacy of clot removal with use of the Hydrolyser thrombectomy catheter in acute massive pulmonary embolism (PE). MATERIALS AND METHODS: Eleven patients (eight women, three men) with a mean age of 61 (range, 37-79) years with acute massive PE underwent percutaneous mechanical thrombectomy (PMT) with use of the Hydrolyser. In four patients with no contraindication, fibrinolysis was performed with use of urokinase at low doses after thrombectomy. RESULTS: Ten patients (90.91%) recovered from massive PE and were discharged within 11 days. The Urokinase Pulmonary Embolism Trial angiographic severity indexes (mean +/- SD) were 14.7 +/- 2.6 and 7.5 +/- 2.7, respectively, before and after thrombectomy (P < .001). Partial arterial pressures of O2 increased from 72.8 mm Hg +/- 16.4 to 93.5 mm Hg +/- 5.6 (P < .005). Pulmonary artery pressure decreased from 45.5 mm Hg +/- 14.2 to 29.5 mm Hg +/- 13.6 after thrombectomy (P < .0001). Calculated by semiquantitative computed analysis, PMT with use of the Hydrolyser removed 74.06% of thrombus +/- 13.46%. One patient developed self limited hemoptysis immediately after thrombectomy. One patient died during the procedure secondary to PE. CONCLUSION: PMT with use of the Hydrolyser is effective and safe in massive PE, resulting in improved hemodynamics and blood oxygenation and decreased pulmonary artery pressure. It offers an alternative to fibrinolysis and surgical thrombectomy. PMID- 11041473 TI - Limb kinking in supported and unsupported abdominal aortic stent-grafts. AB - PURPOSE: The occurrence of kinking of stent-graft limbs depends on the patient's anatomy and the device used. The purpose of this investigation was to determine the rates of limb kinking in supported and unsupported aortic stent-grafts. MATERIALS AND METHODS: The authors performed a retrospective review of patients undergoing placement of either a Guidant Ancure/EGS or Medtronic Talent aortic stent-graft for the treatment of abdominal aortic aneurysm as part of separate phase II and phase III clinical trials. The records of 91 consecutive patients with 149 limbs were reviewed. The type and configuration of each device and any procedure performed specifically relating to limb patency was recorded. An analysis was then performed comparing the rates of kinking in supported and unsupported groups. A review of the literature was also performed. RESULTS: Overall, there was kinking in 18 of 149 limbs (12%). In the supported stent-graft group, 48 bifurcated and 26 aortomonoiliac grafts were placed, with a total of 122 limbs at risk. Six limbs (5%) in five patients required intervention as a result of limb kinking. Stents were placed intraoperatively in two limbs (2%) and postoperatively in four limbs (3%) for thrombosis or severe stenosis. In the unsupported group, 12 bifurcated and three aortomonoiliac grafts were placed, with a total of 27 limbs at risk. Twelve limbs (44%) in eight patients required some type of intervention as a result of limb kinking. Stents were placed intraoperatively in seven limbs (26%) and postoperatively in five limbs (19%) for thrombosis or severe stenosis. Rates of limb kinking were significantly different between the supported and unsupported groups (P < .0001). CONCLUSIONS: The use of supported versus unsupported stent-grafts impacts the occurrence of limb kinking. A direct comparison of the groups suggests that an unsupported stent-graft will be more than 15 times more likely than a supported system to require intervention because of kinking. PMID- 11041474 TI - Influence of radiographic technique and equipment on absorbed ovarian dose associated with uterine artery embolization. AB - PURPOSE: To evaluate the influence of pulsed fluoroscopy (PF), nonpulsed fluoroscopy (NPF), and various fluoroscopic techniques on the absorbed ovarian dose (AOD) associated with uterine artery embolization (UAE) of leiomyomata. MATERIAL AND METHODS: Ovarian location was estimated from preprocedural pelvic magnetic resonance images of 23 patients previously treated by means of UAE. The AOD was measured with thermoluminescent dosimeters (TLD) placed into an anthropomorphic phantom at the determined ovarian location. The following measurements from PF and NPF were obtained: 21.89 minutes of nonmagnified posterior-anterior fluoroscopy, 10 minutes of nonmagnified oblique fluoroscopy, 10 minutes of posterior-anterior magnified fluoroscopy, 10 minutes of combined oblique magnified fluoroscopy, and 47 simulated angiographic exposures. Numbers for nonmagnified posterior-anterior fluoroscopy time and exposure numbers were chosen from the average values from previous UAE procedures. AOD from pulsed and nonpulsed nonmagnified posterior-anterior fluoroscopy was compared to measurements from oblique magnified, posterior-anterior magnified, and oblique fluoroscopy. RESULTS: AOD from NPF was, on average, 1.7 times higher than from PF. When compared with nonmagnified posterior-anterior fluoroscopy, the AOD from oblique magnified fluoroscopy was 1.9 times greater; the AOD from nonmagnified oblique fluoroscopy was 1.1 times greater. The AOD from oblique magnified fluoroscopy was 1.5 times higher on the side closer to the x-ray tube than on the contralateral side. AOD from serial angiographic exposures contributed only less than 7% to the total AOD for the average UAE procedure. CONCLUSIONS: The AOD associated with UAE can best be reduced by limiting fluoroscopy time and the use of oblique or magnified fluoroscopy. Contribution of angiographic exposures to AOD is much less significant. PMID- 11041475 TI - Common femoral artery anastomotic pseudoaneurysm: endovascular treatment with hemobahn stent-grafts. PMID- 11041476 TI - Endovascular treatment of acute carotid blow-out syndrome. PMID- 11041477 TI - Percutaneous stent-graft management of renal artery aneurysms. PMID- 11041478 TI - Direct percutaneous embolization of visceral artery aneurysms: techniques and pitfalls. PMID- 11041480 TI - Repositioning the 12-F over-the-wire Greenfield filter. PMID- 11041479 TI - Multiple pancreaticoduodenal aneurysms: treatment with superior mesenteric artery stent-graft placement and distal embolization. PMID- 11041481 TI - Transjugular intrahepatic portosystemic shunts in the treatment of refractory ascites: results in 48 consecutive patients. AB - PURPOSE: To assess the efficacy, morbidity, and mortality involved in the creation of transjugular intrahepatic portosystemic shunts (TIPS) in the treatment of patients with refractory ascites in Child-Pugh classes B and C. MATERIALS AND METHODS: Forty-eight consecutive patients with refractory ascites were treated with TIPS creation in a tertiary care institution. They were followed for a median of 337 days (range, 3-1376 d). RESULTS: TIPS significantly decreased the portohepatic pressure gradient (20.7 +/- 5.9 mm Hg vs. 6.8 +/- 4.1 mm Hg; P < .0001). Seventy-three percent of patients had complete or partial response. One year after TIPS creation, survival was 73% in Child class B patients and 56% in Child class C patients. Thirteen patients experienced procedural complications (portal vein thrombosis, peritoneal bleeding, acute renal failure, pneumothorax, hemoptysis, spontaneous bacterial peritonitis, and heart failure) and TIPS creation was considered the cause of death in five patients (10.4%). Primary patency was 65% at 3 months and 23% at 1 year, but shunt obstruction was accessible for a second intervention. Ten patients (21%) had de novo encephalopathy after TIPS creation. CONCLUSIONS: This series suggests that TIPS is an effective treatment for refractory ascites; however, it is a challenging procedure and serious complications--usually renal and heart failure- can be seen. A careful selection of patients is mandatory. PMID- 11041482 TI - Effect of the learning process on procedure times and radiation exposure for CT fluoroscopy-guided percutaneous biopsy procedures. AB - PURPOSE: To assess if the learning process associated with computed tomography fluoroscopy (CTF) technology influences procedure and fluoroscopy times for percutaneous biopsy procedures. MATERIALS AND METHODS: Prospective analysis of the initial 250 consecutive patients who underwent percutaneous biopsy with use of a CT scanner equipped with rapid image reconstruction and fluoroscopic capabilities in a 24-month period. All procedures were performed with both continuous and spot fluoroscopic technique, with typical radiation parameters of 50 mA, 120 kV, and a 10-mm-slice thickness. The procedures were all performed by a single experienced interventional radiologist to limit the variables of physician expertise, interventional materials used, and biopsy approach. The subject group was divided into five equal consecutive groups of 50 patients. In each subgroup, the authors recorded mean lesion size, success, and complication rates, as well as mean procedure and fluoroscopy times. RESULTS: The five subgroups were similar patient populations as documented by the absence of statistically significant differences when comparing mean lesion size, procedure success, and complication rates (P > .05; ANOVA test). A statistically significant decrease in mean fluoroscopy (groups 1-5: 50.26 vs 45.24 vs 33.86 vs 32.68 vs 25.8 sec/patient) and mean procedure times (groups 1-5: 30.08 vs 27.9 vs 26.34 vs 25.6 vs 21.6 min/patient) was recorded between the patient subgroups (P < .0001; ANOVA test). CONCLUSION: The learning process associated with CTF technology impacts procedure parameters by decreasing both mean procedure and fluoroscopy times, thereby increasing patient turnover and decreasing radiation exposure to the patient and the operator. PMID- 11041483 TI - Creation of radiopaque thrombi for in vivo experiments. AB - PURPOSE: A number of percutaneous thrombectomy devices are undergoing investigation for treatment of patients with venous thromboembolism. Use of radiopaque thrombus to monitor thrombus delivery and assess thrombectomy has been previously reported. The purpose of this project was to quantitatively test the effect of mixing different ratios of blood and contrast material to facilitate maximum thrombus formation and radiopacity. MATERIALS AND METHODS: The following ratios of blood and contrast material were mixed: 2 mL blood to 8 mL contrast material (ratio = 0.25), 4 mL blood to 6 mL contrast material (ratio = 0.67), 6 mL blood to 4 mL contrast material (ratio = 1.5), and 8 mL blood to 2 mL contrast material (ratio = 4). Contrast material was added at day 0, 3, or 6. Each sample received one of two ionic contrast agents to opacify the clots. At day 14, thrombus mass and opacity were determined. RESULTS: Three combinations of blood and contrast material produced maximum thrombus and radiopacity. These were sodium iothalamate 30% with a ratio of 4 with contrast material added on day 0 and sodium iothalamate 60% with a ratio of 1.5 with contrast material added on day 3 or 6. CONCLUSIONS: When forming radiopaque thrombi, significant differences can result from the ratio of blood to contrast material used. Contrast material type can also affect radiopacity and mass formed. The use of optimal ratios of blood to contrast material should maximize device evaluation with minimal wasting of valuable resources such as test subjects, physician time, and equipment. PMID- 11041484 TI - The square stent-based large vessel occluder: an experimental pilot study. AB - PURPOSE: The purpose of this study is in vitro and in vivo experimental evaluation of a square stent-based vascular occlusion device for large vessels. MATERIALS AND METHODS: Square stent-based large vessel occluders (LVO) 5 mm-50 mm in size were constructed from stainless-steel square stents covered by porcine small intestine submucosa (SIS). The LVOs with two back-side barbs were delivered through a guiding catheter. The LVOs with two back-side barbs and two frontal barbs were front-loaded and delivered coaxially. A pusher with a retention mechanism at its end was used for deployment. In vitro testing for competency was performed with use of a flow model with pressure increases. In an experimental pilot study in seven pigs and five dogs, 16 LVOs were placed into the aorta (n = 4), common iliac artery (n = 2), pulmonary artery (n = 4), and medial sacral artery (n = 6). Four animals received two LVOs in different locations. Angiography was performed before and after placement of each LVO. Animals were followed for as long as 3 months with use of angiography and were then killed for gross and histologic evaluation. RESULTS: In vitro LVOs with two and four barbs were easily collapsed and pushed through or front-loaded into guiding catheters (6-F for a 5-mm occluder, 10-F for a 50-mm occluder). A 20-mm LVO adapted to tubular structures 10-15 mm in diameter, forming polygons 17-18.5 mm in length. In the flow model, LVOs endured pressure increases to 300 mm Hg. In vivo, the LVOs self-expanded and adapted to the vessel without migration in all cases. The locking pusher allowed precise LVO placement and engagement of its barbs into the vessel wall before complete deployment, preventing dislodgment by blood flow. Complete arterial occlusion occurred within 10-20 minutes and arteries remained occluded until the animal was killed in all cases. After 2 months, histologic evaluation revealed replacement of SIS by host tissue and its remodeling with variable fibrocytes, fibroblasts, and some inflammatory cells. Complete endothelialization was seen on both sides of the LVO. CONCLUSION: The SIS LVO is effective and reliable for acute and chronic occlusion in a high flow model in an experimental animal. PMID- 11041485 TI - Sequential activation of transcription factors in lens induction. AB - Since the pioneering work of the early 1900s, the lens has been used as a model system for the study of tissue development in vertebrates. A number of embryological transplantation experiments designed to elucidate the role of tissue interactions in the formation of the lens have led to the proposal of a stepwise determination model. This model has recently been refined through the identification of certain transcription factor genes, which exhibit distinct expression patterns and functional properties in the lens cell lineage. Otx2, Pax6, and Lens1 are induced by the adjacent anterior neural plate and expressed in predifferentiated lens ectoderm. Contact between the optic vesicle and lens ectoderm promotes expression of mafs, Soxs, and Prox1, which are responsible for the initiation of lens differentiation programs including crystallin expression, cell elongation, and cell cycle arrest. Further analysis of the expression and functional characteristics of these transcription factors will allow greater detail when describing the orchestration of genetic programs, which control tissue development from induction to maturation. PMID- 11041486 TI - Cloning and characterization of cDNA for syndecan core protein in sea urchin embryos. AB - The cDNA for the core protein of the heparan sulfate proteoglycan, syndecan, of embryos of the sea urchin Anthocidaris crassispina was cloned and characterized. Reverse transcription-polymerase chain reaction (RT-PCR) was used with total ribonucleic acid (RNA) from late gastrula stage embryos and degenerate primers for conserved regions of the core protein, to obtain a 0.1 kb PCR product. A late gastrula stage cDNA library was then screened using the PCR product as a probe. The clones obtained contained an open reading frame of 219 amino acid residues. The predicted product was 41.6% identical to mouse syndecan-1 in the region spanning the cytoplasmic and transmembrane domains. Northern analysis showed that the transcripts were present in unfertilized eggs and maximum expression was detected at the early gastrula stage. Syndecan mRNA was localized around the nuclei at the early cleavage stage, but was then found in the ectodermal cells of the gastrula embryos. Western blotting analysis using the antibody against the recombinant syndecan showed that the proteoglycan was present at a constant level from the unfertilized egg stage through to the pluteus larval stage. Immunostaining revealed that the protein was expressed on apical and basal surfaces of the epithelial wall in blastulae and gastrulae. PMID- 11041487 TI - Homoiogenetic regulation through the ectoderm on localized expression of the hatching gland phenotype in the head area of Xenopus embryos. AB - Ectoderm pieces explanted from embryos of Xenopus laevis were cultured and examined for differentiation of hatching gland cells, using immunoreactivity against anti-XHE (Xenopus hatching enzyme) as a marker. The anterio-dorsal ectoderm excised from stage 12-13 (mid-late gastrula) embryos developed hatching gland cells. Meanwhile, the posterio-, but not the anterio-dorsal ectoderm from stage 11 (early gastrula) embryos developed these cells, although it is not fated to do so during normogenesis. This hatching gland cell differentiation from stage 11 posterior ectoderm was not affected by conjugated sandwich culture with the mesoderm but was suppressed when explants contained an anterior portion of the ectoderm. Conjugated cultures of anterior and posterior portions of the ectoderm in various combinations indicated that differentiation of hatching gland cells from stage 11 posterior and stage 12 anterior portions was suppressed specifically by stage 11 anterior ectoderm. Northern blot analyses of cultured explants showed that XHE was expressed in association with XA-1, suggesting its dependence on the anteriorized state. These results indicate that the planar signal(s) emanating from stage 11 anterior ectoderm participates in suppression of the expression of the anteriorized phenotype so that an ordered differentiation along the anteroposterior axis of the surface ectoderm is accomplished. PMID- 11041488 TI - Activity of the medaka translation elongation factor 1alpha-A promoter examined using the GFP gene as a reporter. AB - The translation elongation factor 1alpha (EF-1alpha) is known to have several isoforms, which are expressed in a tissue- and stage-specific manner. Two genes encoding EF-1alpha exist per haploid genome in the medaka. In the present study, the promoter activity of the 5'-flanking region of the medaka EF-1alpha-A gene, an isoform of EF-1alpha, was characterized using transgenic techniques. First, using CAT gene as a reporter, it was revealed that about 1.8 kbp 5'-flanking sequence from the transcription initiation site of EF-1alpha-A was sufficient for high-level promoter activity. Second, the green fluorescent protein (GFP) gene fused to this region was introduced into medaka eggs using the microinjection method. Three germline transgenic individuals (one male and two female) were mated with non-transgenic medaka to obtain F1 offspring. In the case of embryonic and adult F1 transgenic individuals, GFP fluorescence was observed in almost all the tissues examined (e.g. kidney, liver, heart, gill, ovary, and testis), except for the skeletal muscle. In the case of F2 transgenic embryos derived from F1 transgenic males and non-transgenic females, the fluorescence was observed from the early gastrula stage. On the other hand, in the case of F2 transgenic embryos derived from F1 transgenic females and non-transgenic males, the fluorescence was observed even at the 1-cell stage, suggesting that this region is transcriptionally active during oogenesis. The usefulness of the EF-1alpha-A promoter as a tool for introducing foreign proteins into oocytes is discussed. PMID- 11041489 TI - Initial analysis of immunochemical cell surface properties, location and formation of the serotonergic apical ganglion in sea urchin embryos. AB - In the present study it was found that serotonergic apical ganglion (SAG)-forming cells in plutei of the sea urchin, Hemicentrotus pulcherrimus, possessed a characteristic pear shape with broad apical sides and a pointed basal side in the acron epithelium. The basal side extended axons through the space between the epithelium and the basal lamina toward the midline of the embryo that aligned parallel to the embryonic anteroposterior axis. Serotonergic apical ganglion forming cells had epithelial cell surface-specific proteins on their entire surface. The SAG in 4-arm plutei was composed of a 4-cell trunk region that aligned at right angles to the embryonic anteroposterior axis, and forked into two branches of one to two cells at both ends. Two branches extended toward the oral and the other two toward the aboral region, respectively. Double-stained immunohistochemistry using antiserotonin antibodies and oral ectoderm-specific anti-Ecto V monoclonal antibody or aboral ectoderm-specific anti-Ars antibodies indicated that SAG was in the aboral ectoderm region. Serotonergic apical ganglion cells were first detected in late gastrulae and increased in number rapidly between 36 and 48 h after fertilization, and then slowly afterwards. A 5 bromo-2-deoxyuridine incorporation study indicated that none of the increased SAG cells were in the S phase during the aforementioned period, suggesting that SAG cells do not proliferate by cell division, but acquire the property in particular cells by transdifferentiation using a mechanism that has yet to be elucidated. PMID- 11041490 TI - Effects of rat Axin domains on axis formation in Xenopus embryos. AB - Wnt signaling plays an important role in axis formation in early vertebrate development. Axin is one Wnt signaling regulator that inhibits this pathway. The effects of the injection of mRNA of several rat Axin (rAxin) mutants on axis formation in Xenopus embryos were examined. It was found that rAxin mutants containing only a regulation of G-protein signaling (RGS) domain fragment or with deletion of the RGS domain induced axis formation. Because the RGS domain is a major adenomatous polyposis coli gene product (APC)-binding domain, APC association with glycogen synthase kinase 3beta (GSK3beta) on the Axin molecule may be important in inhibition of axis formation. The ventralizing activities of wild-type rAxin and a mutant in which the Dishevelled and Axin (DIX) domain was deleted (deltaDIX mutant) were examined. Histological examination and gene expression revealed that the ventralizing activity of the deltaDIX mutant was weaker than that of wild-type rAxin. This finding suggests that the C-terminus of rAxin contributes to the inhibition of Wnt signaling in Xenopus embryos. Furthermore, an rAxin mutant that contained both the RGS and GSK3beta-binding domains affected both the dorsal and ventral sides of blastomeres, mediated ectodermal fate and induced expansion of notochord and/or endoderm, but did not induce axis formation. PMID- 11041491 TI - Regulative specification of ectoderm in skeleton disrupted sea urchin embryos treated with monoclonal antibody to Pl-nectin. AB - Pl-nectin is a glycoprotein first discovered in the extracellular matrix (ECM) of Paracentrotus lividus sea urchin embryo, apically located on ectoderm and endoderm cells. The molecule has been described as functioning as an adhesive substrate for embryonic cells and its contact to ectoderm cells is essential for correct skeletogenesis. The present study was undertaken to elucidate the biochemical characteristics of Pl-nectin and to extend knowledge on its in vivo biological function. Here it is shown that the binding of mesenchyme blastula cells to Pl-nectin-coated substrates was calcium dependent, and reached its optimum at 10 mM Ca2+. Perturbation studies using monoclonal antibody (McAb) to Pl-nectin, which prevent ectoderm cell-Pl-nectin contact, show that dorsoventral axis formation and ectoderm differentiation were retarded. At later stages, embryos recovered and, even if growth and patterning of the skeleton was greatly affected, the establishment of dorsoventral asymmetry was reached. Similarly, the expression of specific ectoderm and endoderm territorial markers was achieved, although occurring with some delay. Endoderm differentiation and patterning was not obviously affected. These results suggest that both endoderm and ectoderm cells have regulative capacities and differentiation of territories is restored after a lag period. On the contrary, failure of inductive differentiation of the skeleton cannot be rescued, even though the ectoderm has recovered. PMID- 11041492 TI - Direct molecular interaction of a conserved yolk granule protein in sea urchins. AB - The regulation of yolk storage in oocytes and subsequent utilization in embryos is critical for embryogenesis. In sea urchins, the major yolk protein is made in the intestines, transported to the ovaries and accumulated in developing oocytes within membrane-bound vesicles comprising approximately 10% of the mass of an egg. Here, a non-yolk protein that accumulates specifically in yolk granules is reported. This protein was identified by cDNA cloning and, by use of antibodies to the recombinant protein, it was shown that this molecule is stored selectively in yolk granules of oocytes and embryos. No accumulation was seen in the accessory cells, testis, or intestines. In situ ribonucleic acid (RNA) hybridizations showed that the transcript accumulated only in oocytes, and was more highly concentrated in young oocytes. However, later in oogenesis, the messenger ribonucleic acid (mRNA) levels decreased significantly so that no signal was detectable in mature haploid eggs or at any later stage in development. However, by immunofluorescence and western blot analysis, the 30 kDa band was present throughout development. The predicted sequence of this protein shows that it is a member of the bep, HLC-32, EBP family of sea urchin proteins, but as it does not accumulate at the cell surface, nor in the hyaline layer in the two species studied here, as do other members of the family, it has been referred to as YP30 (30 kDa protein of the yolk platelet). To address its potential function, yeast two-hybrid analysis was performed to screen for proteins that potentially interact with YP30. It was found that it binds itself, and forms strongly interacting dimers. It is hypothesized that YP30 participates in the packaging and storage of major yolk protein during oogenesis, or in the utilization of the major yolk protein in development. PMID- 11041493 TI - Pamlin-induced tyrosine phosphorylation of SUp62 protein in primary mesenchyme cells during early embryogenesis in the sea urchin, Hemicentrotus pulcherrimus. AB - Ingression of primary mesenchyme cells (PMC) is associated with the encounter of basal lamina including pamlin. It was found that sea urchin embryos have a protein that binds antihuman focal adhesion kinase (FAK) antibodies, yet it has a 62 kDa homo-dimeric structure. Thus, this protein was distinctive from known FAK, and was named SUp62. In mesenchyme blastulae, one of the subunits increased its apparent molecular mass slightly but distinctively, then restored the original molecular mass in early gastrulae. This temporal and stage-specific shifting of the molecular mass was associated with the occurrence of tyrosine phosphorylation of a subunit that did not increase the apparent molecular mass. Herbimycin A induced the hyperphosphorylation of tyrosine residues of SUp62, and inhibited the occurrence of molecular mass shifting. Immunohistochemistry showed a strong positive signal of SUp62 and phosphotyrosine in PMC. Herbimycin A also severely but reversibly inhibited PMC dissociation, migration and gastrulation. Tyrosine phosphorylation of SUp62 was induced when PMC were incubated with pamlin in vitro, and it was initiated within 10 min after onset of the incubation. It reached its peak in 1 h, and declined gradually in the next 1 h, indicating that pamlin-induced tyrosine phosphorylation of SUp62 occurs closely associated with acquiring PMC migration activity. PMID- 11041494 TI - A novel role of differentiation-inducing factor-1 in Dictyostelium development, assessed by the restoration of a developmental defect in a mutant lacking mitogen activated protein kinase ERK2. AB - It has been previously reported that the differentiating wild-type cells of Dictyostelium discoideum secrete a diffusible factor or factors that are able to rescue the developmental defect in the mutant lacking extracellular signal regulated kinase 2 (ERK2), encoded by the gene erkB. In the present study, it is demonstrated that differentiation-inducing factor-1 (DIF-1) for stalk cells can mimic the role of the factor(s) and the mechanism of the action of DIF-1 in the erkB null mutant is also discussed. The mutant usually never forms multicellular aggregates, because of its defect in cyclic adenosine monophosphate (cAMP) signaling. In the presence of 100 nM DIF-1, however, the mutant cells formed tiny slugs, which eventually developed into small fruiting bodies. In contrast, DIF-1 never rescued the developmental arrest of other Dictyostelium mutants lacking adenylyl cyclase A (ACA), cAMP receptors cAR1 and cAR3, heterotrimeric G-protein, the cytosolic regulator of ACA, or the catalytic subunit of cAMP-dependent protein kinase (PKA-C). Most importantly, it was found that DIF-1 did not affect the cellular cAMP level, but rather elevated the transcriptional level of pka during the development of erkB null cells. These results suggest that DIF-1 may rescue the developmental defect in erkB null cells via the increase in PKA activity, thus giving the first conclusive evidence that DIF-1 plays a crucial role in the early events of Dictyostelium development as well as in prestalk and stalk cell induction. PMID- 11041496 TI - Competitive exclusion treatment reduces the mortality and fecal shedding associated with enterotoxigenic Escherichia coli infection in nursery-raised neonatal pigs. AB - We have previously reported that the administration of a competitive exclusion culture (PCF-1), derived from the cecal microflora of a young, healthy pig and maintained in a continuous flow fermentation system to neonatal pigs resulted in a decrease in the incidence of fecal shedding and cecal colonization by Salmonella choleraesuis in pigs at weaning. In the present experiment, we describe the effects of the administration of a derivative of the PCF-1 culture, RPCF, against an enterotoxigenic E. coli infection in neonatal pigs raised off sow. The administration of RPCF at 12 and 24 hours after birth resulted in significant (P < 0.05) reductions in mortality, incidence of fecal shedding, and in gut colonization by E. coli when compared to control values. The RPCF reduced mortality from 17.5%, observed in untreated pigs, to 4.4% in RPCF-treated pigs. Fecal shedding of E. coli was reduced significantly (P < 0.05) in RPCF-treated pigs between Days 1 and 3 post-challenge. These results indicate that the RPCF culture is effective against one of the most important causes of neonatal scours (E. coli infections) in piglets. PMID- 11041495 TI - Xenotransplantation and the potential risk of xenogeneic transmission of porcine viruses. AB - The clinical success of allotransplantation and the shortage of donor organs have led to a proposal for the use of animal organs as alternative therapeutic materials for humans. In that regard, swine are preferable to non-human primates as a source of donor organs. While applications for clinical trials for xenotransplantation have not yet been received in Canada, several trials have already been authorized in the United States. A major concern, however, is the potential for xenogeneic transmission of viruses from animals to humans via organ, tissue, or cellular transplantation or via ex vivo exposure of humans to porcine biologic materials. Xenotransplantation allows viruses to bypass the normal immunological defense mechanisms of the recipient. Furthermore, the use of immunosuppressive drugs following transplantation may facilitate the xenogeneic transmission of zoonotic agents. Of porcine viruses, swine hepatitis E virus does not cause any clinical symptoms in the natural host but is a likely zoonotic agent that can infect humans and cause hepatitis. Porcine circovirus type 1 is prevalent in swine populations with no known association with clinical disease, while circovirus type 2 causes post-weaning multi-systemic wasting syndrome. Porcine endogenous retrovirus is integrated into the host chromosomes while porcine cytomegalovirus undergoes latent infection. Two additional porcine herpesviruses have recently been identified in swine and have been named porcine lymphotrophic herpesviruses. These herpesviruses can potentially become reactivated in human recipients after xenotransplantation. All in all, there are a number of viruses in swine that are of primary concern to screen and eliminate from xenotransplantation protocols. Epidemiology and the current knowledge on xenogeneic risk of these viruses are discussed. PMID- 11041497 TI - Yohimbine ameliorates the effects of endotoxin on gastric emptying of the liquid marker acetaminophen in horses. AB - The effect of yohimbine pretreatment on gastric emptying of a liquid marker in horses was evaluated by measuring serum concentrations of acetaminophen. Gastric emptying was determined in normal, fasted horses, in horses given endotoxin (E. coli 055 B5; 0.2 microg/kg) intravenously, and in horses given yohimbine (0.25 mg/kg, IV, over 30 minutes) plus endotoxin. Acetaminophen (20 mg/kg) was given by stomach tube 15 minutes after the endotoxin infusion. Blood samples for acetaminophen analysis were collected, and time to reach the peak serum concentration (Tmax), the maximum serum concentration (Cmax) and the area under the acetaminophen serum concentration versus time curve (AUC) were determined for each treatment group. Endotoxin significantly increased Tmax, indicating a profound delay in gastric emptying and yohimbine pretreatment significantly (P < or = 0.05) prevented this effect. PMID- 11041498 TI - An evaluation of chemical arthrodesis of the proximal interphalangeal joint in the horse by using monoiodoacetate. AB - The use of monoiodoacetate (MIA) for arthrodesis of the proximal interphalangeal joint (PIJ) and the effect of exercise on the degree of fusion were investigated. Eight horses received 3 injections (Weeks 0, 3, 6) of MIA (2 mL; 60 mg/mL) into the right or left front PIJ. Peri-operatively, the horses received phenylbutazone, butorphanol, and abaxial sesamoidean nerve blocks to relieve pain. During the study, the horses were monitored for general health, lameness, and swelling around the injection area. Radiographs were taken biweekly to evaluate bony fusion. Horses were randomly divided into non-exercised and exercised groups. Exercise consisted of 20 minutes of trotting on a treadmill (4 m/s), 3 days per week for 13 weeks. The horses were euthanized at 24 weeks. Slab sections of the PIJ were evaluated grossly and radiographically for bony fusion. Histologic examinations were performed to evaluate articular cartilage. Three horses were excluded from the study after developing soft tissue necrosis around the injection site, septic arthritis, and necrotic tendinitis. The remaining horses remained healthy, developed a grade 1 to 4 lameness with minimal to severe swelling in the PIJ region. All 5 horses showed radiographic evidence of bony fusion, however, no fusion was present when injected joints were examined on postmortem examination. Histologic examination revealed thinning of the cartilage, diffuse necrosis of chondrocytes, with the calcified zone intact. Subjectively, exercise did not influence the degree of cartilage destruction. Based on this study, chemical arthrodesis cannot be advocated in clinical cases because of the high complication rate and lack of bony fusion. PMID- 11041499 TI - Blood acid-base and serum electrolyte values in red deer (Cervus elaphus). AB - Acid-base, serum electrolyte, plasma protein, and packed cell volume (PCV) values were determined in venous blood samples from 30 red deer (Cervus elaphus) of both sexes showing no clinical signs of disease. The animals were 5 months of age and kept on pasture in the Valley of Mexico, at an altitude of 2450 m. Blood samples were collected without sedation. Mean blood values were: pH 7.411 +/- 0.041, pCO2 37.7 +/- 4.4 mmHg, base excess 0.7 +/- 3.2 mmol/L, actual bicarbonate 24.3 +/- 3.1 mmol/L, total CO2 25.3 +/- 3.2 mmol/L and anion gap 23.5 +/- 5.5 mmol/L. Mean serum electrolyte levels were: Na+ 142.3 +/- 2.5 mmol/L, Cl- 100.5 +/- 2.3 mmol/L, and K+ 7.03 +/- 1.03 mmol/L. Plasma protein and PCV values were 60.0 +/- 6.6 g/L and 0.47 +/- 0.05 L/L, respectively. Blood values determined in this study can be considered reference data for health control and disease diagnosis. PMID- 11041500 TI - Infection of Bergmann glia in the cerebellum of a skunk experimentally infected with street rabies virus. AB - Rabies virus is a highly neuronotropic virus and glial cell infection is not prominent in the central nervous system (CNS). Paraffin-embedded tissues from the cerebella of skunks experimentally infected with either a skunk salivary gland isolate of street rabies virus or the challenge virus standard (CVS) strain of fixed rabies virus were examined with immunoperoxidase staining for rabies virus antigen by using an anti-rabies virus nucleocapsid protein monoclonal antibody. A skunk infected with street rabies virus showed prominent infection of Bergmann glia. Although infected Purkinje cells were observed, they usually demonstrated a relatively small amount of antigen in their perikarya. A CVS-infected skunk showed many intensely labeled Purkinje cells and a relatively small number of infected Bergmann glia. These findings indicate that although rabies virus is a highly neuronotropic virus, street rabies virus strains do not always demonstrate strict neuronotropism in the central nervous system. PMID- 11041501 TI - Naturally occurring lesions of the uterine tube in sheep and serologic evidence of exposure to Chlamydophila abortus. AB - The uterine tubes from 405 ewes, collected at an abattoir, were assessed grossly and microscopically for abnormalities that correlated with serological evidence of exposure to Chlamydophila abortus. Gross lesions were found in 41 ewes and 86 had microscopic lesions. Enzyme immunoassay (EIA) of serum was used as an indication of exposure of individual ewes to C. abortus; 52 were found to be positive. Chi-squared analysis indicated no association between EIA-positive animals and lesions of the uterine tube. PMID- 11041502 TI - Longitudinal evaluation of CD4+ and CD8+ peripheral blood and mammary gland lymphocytes in cows experimentally inoculated with Staphylococcus aureus. AB - Staphylococcus aureus is a major pathogen associated with mastitis, a disease affecting both women and dairy cows. The longitudinal profiles of bovine peripheral blood and mammary gland lymphocyte phenotypes in response to S. aureus induced mastitis were investigated in dairy cows. Increased percentage of CD4 lymphocytes in the mammary gland between 1 and 8 days post-inoculation, increased milk CD4 protein density per cell between 1-8 days post-inoculation, and a statistically significant negative correlation between post-inoculation bacterial counts in milk and blood lymphocyte CD4 protein density were found. Together with blood and milk leukocyte counts, the milk lymphocyte CD4/CD8 ratio and the milk lymphocyte CD4 protein density were more informative indicators than milk somatic cell counts and bacteriology for identification of early vs. late inflammatory phases. These findings suggest that CD4+ lymphocytes play a protective role in the early stages of S. aureus-induced mastitis. PMID- 11041503 TI - Tilmicosin does not inhibit interleukin-8 gene expression in the bovine lung experimentally infected with Mannheimia (Pasteurella) haemolytica. AB - The expression of the interleukin-8 (IL-8) gene was examined by in situ hybridization in lung tissues from calves experimentally infected with Mannheimia (Pasteurella) haemolytica and treated with tilmicosin. Interleukin-8 mRNA expression was detected in alveolar areas, particularly along interlobular septa, in the lumen, and in the epithelial cells of some bronchioles. In lesional lung tissues from animals that had received tilmicosin, we found large areas with limited inflammation. There was no staining for IL-8 mRNA in these areas. In contrast, in strongly inflamed areas, the same patterns and intensities of staining for IL-8 mRNA were detected in tilmicosin- and sham-treated animals. We conclude that tilmicosin does not affect the expression of IL-8 mRNA in tissue showing microscopic signs of inflammation. Together with previous reports, this supports the view that the pro-apoptotic properties of tilmicosin on neutrophils do not compromise the host defense mechanisms required to control the infection. PMID- 11041504 TI - Achondroplastic dog breeds have no mutations in the transmembrane domain of the FGFR-3 gene. AB - One of the most common skeletal affections in humans is achondroplasia, a short limbed dwarfism that is, in most cases, caused by mutations in the transmembrane domain of the fibroblast growth factor receptor-3 (FGFR-3) gene. Due to the lack of sufficient radiological, genetic, and molecular studies, most types of skeletal anomalies in dogs are classified as achondroplasia. To initiate the molecular characterization of some osteochondrodysplastic dog breeds, we obtained the DNA sequence of the transmembrane domain of the FGFR-3 gene from the dachshund, basset hound, bulldog, and German shepherd dogs. All 4 breeds showed no mutation in the evaluated region. This indicates that the mutation responsible for the osteochondrodysplastic phenotype in the tested dog breeds lies either elsewhere in the FGFR-3 gene or in other ones involved in the formation and development of endochondral bone. PMID- 11041505 TI - The effect of stanozolol on 15nitrogen retention in the dog. AB - The objective of the study was to determine the influence of either oral or intramuscular administration of stanozolol on nitrogen retention in dogs by using a non-invasive 15N-amino acid tracer technique. Ten healthy, intact, adult male sled dogs received either stanozolol tablets, 2 mg/dog PO, q12h, for 25 days (Group 1, n = 5) or an intramuscular injection of 25 mg of stanozolol on Days 7, 14, 21, and 28 (Group 2, n = 5). A 15N amino acid (5.27 mmol) was infused intravenously into each dog on Day 0 (before stanozolol treatment) and on Day 31 (after stanozolol treatment). Urine was collected by catheterization from each animal 3 times daily for 3 consecutive days. The 15N-urea enrichment in urine was determined by high-resolution mass spectrometry and the total amount of urea in the urine was determined. Both oral and injectable stanozolol resulted in significant (P < 0.05) increases in amino acid nitrogen retention compared to pretreatment values. Oral stanozolol increased nitrogen retention from 29.2 +/- 8.2% to 50.3 +/- 9.2%, while stanozolol injection increased nitrogen retention from 26.6 +/- 9.9% to 67.0 +/- 7.5%. The response to intramuscular administration was significantly greater than the response to the oral dosing regime. Stanozolol increases amino acid nitrogen retention in dogs, as has been previously observed in rats. This action of stanozolol may be beneficial in dogs under stress of surgical trauma and chronic disease. PMID- 11041507 TI - Pacing induced sustained atrial fibrillation in a pony. AB - A transvenous, screw-in electrode was implanted in the right atrium of a healthy pony and connected with an implantable pulse generator programmed to deliver bursts of electrical stimuli to the atrium. Initially, cessation of burst pacing resulted in short (less than 1 minute), self-terminating episodes of atrial fibrillation. As burst pacing continued, the episodes of induced atrial fibrillation became longer. After 3 weeks of continuous atrial pacing, atrial fibrillation became sustained (56 hours). This model of pacing induced atrial fibrillation can be used to study the mechanisms leading to atrial fibrillation, its perpetuation and therapy. Our preliminary observations support the concept that once atrial fibrillation starts, it sets up changes in the electrical characteristics of the atrium that favor its own perpetuation. PMID- 11041506 TI - Propofol or thiopentone as induction agents in romifidine-sedated and halothane N2O-anesthetized dogs: a preliminary study. AB - The objective of this paper was to evaluate the use of romifidine as a premedicant in dogs before general anesthesia induced with propofol or thiopentone and maintained with halothane-N2O. Fifteen healthy dogs were anesthetized twice. Each dog received, as preanesthetic protocol, atropine (10 microg/kg, IM) and romifidine (40 microg/kg, IM); induction was delivered with propofol or thiopentone and anesthesia was maintained with halothane and N2O for 1 h. Some cardiovascular and respiratory variables and recovery times were recorded. Induction doses of propofol or thiopentone and the percentage of halothane necessary for maintaining anesthesia were also registered. Thiopentone as an induction agent is more respiratory depressive but is less hypotensive than propofol. Thiopentone reduces further the percentage of halothane necessary for maintaining the anesthesia. However, the quality of recovery is poorer, as the time to extubation is longer and the dogs occasionally had a violent recovery. The combination of romifidine, atropine, propofol, halothane, and N2O appears to be an effective combination for inducing and maintaining general anesthesia in healthy dogs. PMID- 11041508 TI - Origin and persistence of the mitochondrial genome. AB - The mitochondrial genome comprises a circular, histone-free 'chromosome' of 16.6 kb of DNA, present in one or more copies in every mitochondrion. This chromosome has been tightly conserved for more than half a billion years, coding in every multicellular animal so far investigated, both vertebrate and invertebrate: (i) the same 13 protein subunits required for oxidative phosphorylation; (ii) a component of each of the two mitochondrial ribosome subunits; and (iii) the 22 transport RNAs present within the mitochondrion. Exons on the circle are tightly packed, with no spacing introns. Mitochondrial DNA is histone-free, has limited repair ability, and has a relatively high mutation-fixation rate. Inheritance is cytoplasmic and maternal, with epidemiological evidence (namely the familial distribution of polymorphisms) indicating that recombination with mtDNA of paternal origin is exceedingly rare. Thus the maintenance and evolution of mtDNA (its remarkably successful symbiotic persistence with the nuclear genome) has been essentially asexual. The machinery for homologous recombination is present in mitochondria of at least some species, however, and it might be surprising if it did not occur between circles in some circumstances. By bringing together the fields of mitochondrial biochemistry, evolutionary genetics, reproductive physiology, and neuromuscular medicine in focusing on the inheritance of normal and abnormal human mtDNA, we can hope to better understand the forces behind this genome's inheritance and what might be required of ovarian function to satisfy its accurate persistence over millions of years. Clinically we can hope also for a better understanding of ooplasmic factors in human fertility and in the wide manifestations of mitochondrial genomic disease. PMID- 11041509 TI - Transcription and replication of mitochondrial DNA. AB - The physical isolation of mammalian mitochondrial DNA (mtDNA) over 30 years ago marked the beginning of studies of its structure, replication and the expression of its genetic content. Such analyses have revealed a number of surprises: novel DNA structural features of the circular genome such as the displacement loop (D loop); multiple sized and deleted forms of the circular genome; a minimal set of mitochondrially encoded rRNAs and tRNAs needed for translation; a bacteriophage like, nuclear-encoded mitochondrial RNA polymerase for transcription; and a direct linkage between transcription and the commitment to replication of the leading mtDNA strand that centres on the nuclear encoded mitochondrial transcription factor A. One of the more recent revelations is the existence, near the D-loop, of an atypical, stable RNA-DNA hybrid (or R-loop) at the origin of mammalian leading-strand DNA replication, composed of the parent DNA strands and an RNA transcript. In mammalian mitochondrial systems, all of the proteins known to be involved in DNA replication are encoded in the nucleus. Thus alterations and deficiencies in mtDNA replication must arise from mutations in mtDNA regulatory sequences and nuclear gene defects. Further studies of the relationships between nuclear-encoded proteins and their mtDNA target sequences could result in strategies to manipulate genotypes within cellular mtDNA populations. PMID- 11041510 TI - Genetic control of oxidative phosphorylation and experimental models of defects. AB - Energy in the form of ATP is continually produced by all cells for normal growth and function. Anaerobic glycolysis can provide enough ATP for some cells, but energetic cells such as cardiomyocytes and neurons require a more efficient ATP supply, which can only be provided by mitochondrial oxidative phosphorylation. Invented by bacteria that became symbiotically associated with other bacteria to form eukaryotic cells billions of years ago, oxidative phosphorylation carries with it a genetic legacy that is unique. The mitochondrial oxidative phosphorylation complexes are assembled from protein subunits encoded by both the mitochondrial genome (mtDNA) and the nuclear genome (nDNA, located in the chromosomes). The mtDNA is a remnant genome of the bacterial progenitor of mitochondria, and (unlike the biparental diploidy that characterizes the nuclear genome) is present in thousands of copies per cell, is replicated through life, and is inherited (cytoplasmically) only from the female parent. Oxidative phosphorylation comprises five multimeric enzyme complexes that act as a redox pathway, passing electrons from oxidizable intermediates produced by the metabolism of food to molecular oxygen in the mitochondrial matrix, while producing an electrochemical gradient by pumping protons into the intermembranal space. The proton (hydrogen ion) gradient across the inner mitochondrial membrane is used by the H+-transporting ATP synthase to produce ATP from ADP and inorganic phosphate, with the protons released into the mitochondrial matrix then combining with electronated oxygen to form water. Many of the details regarding the control of the synthesis of oxidative phosphorylation enzyme complexes remain to be elucidated. Transmitochondrial cell culture systems have been developed so that defective oxidative phosphorylation can be studied in a controlled nuclear background. Such systems may soon enable the development of mtDNA 'knockout' mice in order to better model mtDNA transmission and mitochondrial disease. PMID- 11041511 TI - Genetic defects causing mitochondrial respiratory chain disorders and disease. AB - Genetic mitochondrial defects of the respiratory chain show marked phenotypic variability. Laboratory diagnosis is complicated and includes biochemical screening tests, tissue histopathology, functional enzyme studies, and molecular tests where available. Normal respiratory chain function necessitates the co ordinated expression of over 100 different gene loci, and the interaction of two genetic systems, the nuclear and mitochondrial genomes. Thus genetic counselling for the mitochondrial disorders is extremely challenging. In this review, the classes of mitochondrial and nuclear defects that give rise to functional abnormalities of the mitochondrial respiratory chain are discussed, with specific instructive examples described in some detail. PMID- 11041512 TI - Organismal effects of mitochondrial dysfunction. AB - Mitochondrial disease can lead to clinical abnormalities in any organ system. Both inherited and spontaneous disorders are known. The spontaneous forms can occur as a mitochondrial DNA (mtDNA) mutation early in embryogenesis or, later in life, as somatic mutations that accumulate with age. The inherited forms may arise from any of >100 characterized mutations in mtDNA or from >200 nuclear gene defects that affect proteins required for mitochondrial function. Most dividing cells survive and interact normally despite their mitochondrial defects. Thus post-mitotic, terminally differentiated cells are preferentially affected in mitochondrial disease. This review emphasizes cellular metabolic co-operation and the structural and biochemical diversity of mitochondria as the framework for understanding the clinical spectrum of mitochondrial disease. The principles of the mitochondrial clinical assessment scale I (MCAS-I) are presented to assist in the development of diagnostic spectra of mitochondrial disease. PMID- 11041514 TI - Morphological correlates of mitochondrial dysfunction in children. AB - Morphological studies have traditionally played a major role in the study of adults with suspected mitochondrial diseases. Here we review their role in the investigation of paediatric patients. The morphological changes may be macroscopic, such as developmental abnormalities of the brain in pyruvate dehydrogenase deficiency, including ectopic inferior olives and the absence of corpus callosum and pyramids. Other changes are histological, such as rarefaction of the neuropil and endothelial prominence in Leigh syndrome, and spongiosis with neuronal loss and gliosis in Alpers disease. The ragged-red fibres typical of mitochondrial disease in adults are only rarely seen in skeletal muscle biopsies from children. On the other hand, dramatic ultrastructural changes involving the mitochondria may be seen in many organs, including the liver, heart and intestine. In Alpers and lethal infantile mitochondrial diseases, the hepatocytes show marked accumulation of small droplets of lipid alternating with densely packed mitochondria with pale matrix and loss of granules. These changes are associated with a marked decrease in respiratory chain enzyme activity in the liver, often without similar decrease in the skeletal muscle or fibroblasts. Enlarged mitochondria with concentric cristae are prominent in the cardiac myocytes in Barth syndrome. For the assessment of children with a suspected disorder of mitochondrial dysfunction, detailed morphological studies of the brain (at autopsy) and of biopsies (especially of the liver), including ultrastructural assessment of the mitochondria, can be a very useful preliminary investigation. The findings should then be correlated with the clinical features and used as a guide for further biochemical and molecular studies, preferably on multiple tissues. PMID- 11041513 TI - Practical problems in detecting abnormal mitochondrial function and genomes. AB - Mitochondrial respiratory chain dysfunction causes a wide range of primary diseases in adults and children, with highly variable organ involvement. Diagnosis involves weighing evidence from a number of sources, including the clinical presentation, metabolic measurements in vivo, imaging studies, analysis of respiratory chain function or enzyme activities in vitro, studies of mitochondrial morphology after biopsy, and mitochondrial (mt) DNA mutation analysis. Irrespective of the category of the information, it can be difficult to determine whether abnormal results are due to primary defects of the respiratory chain or to practical problems that complicate the diagnostic methodology. This review describes six sources of such problems: genetic complexity, tissue and temporal variation, methodological limitations, secondary effects, logistical issues, and questions of interpretation. When these issues are all addressed, a reliable categorization of the diagnosis as definite, probable, or possible respiratory chain defect becomes possible. PMID- 11041515 TI - In-vitro genetic modification of mitochondrial function. AB - Defects of mitochondrial (mt) DNA cause a diverse group of incurable, progressive diseases that often lead to severe disability and premature death. Most patients with pathogenic mtDNA defects have a mixture of mutant and wild-type mtDNA (heteroplasmy), and the clinical defect is only expressed when the percentage of mutant mtDNA exceeds a critical threshold. Since mtDNA is continually replicating and being turned over, we have proposed an approach to the treatment of these disorders that utilizes sequence-specific antigenomic peptide nucleic acids (PNAs) to hybridize and specifically inhibit the replication of mutant mtDNA under physiological conditions. By allowing the selective propagation of wild type molecules, it may be possible to correct the cellular biochemical defect and to prevent the progression of disease. This paper summarizes the experimental progress in this area, including the cellular uptake of PNA molecules and their import into mitochondria both in vitro and in cell culture by the addition of a nuclear-encoded mitochondrial targeting sequence. The possibilities of extending this strategy to the treatment of mtDNA deletion disorders are discussed. PMID- 11041516 TI - Regulation of mitochondrial DNA copy number during spermatogenesis. AB - The nuclear genome is physically compacted during spermatogenesis by replacing histones with protamines and transition proteins. This altered nuclear protein context may make gene regulation at the transcriptional level less efficient and could explain why post-transcriptional regulation is prominent in haploid male germ cells. Mitochondria and mitochondrial (mt) DNA are maternally inherited, whereas the transmission of paternal mtDNA is blocked in mammals. The paternal mtDNA enters the oocyte but is no longer detectable in the preimplantation embryo. Several mechanisms could be responsible for preventing the transmission of paternal mtDNA, including the down-regulation of mtDNA copy number during spermatogenesis, specific elimination of paternal mitochondria in fertilized oocytes, and the suspension of mtDNA replication in the fertilized oocyte. It is the first of these that is the subject of the present review. Mitochondrial transcription factor A (mtTFA, or Tfam) is a key regulator of mtDNA copy number in mammals. Germ cell-specific Tfam transcript isoforms are expressed during spermatogenesis in mice and humans. These alternative Tfam transcript isoforms have a structure that could prevent protein translation; their expression coincides with down-regulation of the mitochondrial Tfam protein values. We propose that this down-regulation of mitochondrial Tfam protein levels in turn down-regulates mtDNA copy number during mammalian spermatogenesis. PMID- 11041517 TI - Fertilization and elimination of the paternal mitochondrial genome. AB - With rare exceptions, mammalian mitochondria are inherited through the female. This probably serves to minimize lethal cytoplasmic gene competition and to prevent the inheritance of sperm mitochondrial DNA that has been subject to degradation by free radicals. In general, organisms are intolerant of mitochondrial heteroplasmy and, when this occurs in humans, it frequently presents as progressive and lethal bioenergetic or neurological disease. The mitochondria of spermatozoa are specifically destroyed by proteolysis in early embryonic development, in mice at the 4- to 8-cell transition. While there are concerns in human assisted reproduction that microinjection of abnormal or immature sperm cells could lead to lasting harm in the offspring through transmission of abnormal mitochondria, there is no clinical evidence to support this. There is more potential for harm through attempts to 'rescue' poor quality oocytes by cytoplasmic or nuclear transfer, as it is not currently possible to control the final fate of the donated mitochondria in relation to nuclear mitochondrial interactions or the embryonic axes. Moreover, the balance between nuclear and mitochondrial genes and the role of cytoplasmic factors in epigenesis are still poorly understood. The future challenge for biologists is to comprehend the nature of the selective destruction of paternal mitochondria, as it appears to be a species-specific recognition phenomenon. PMID- 11041518 TI - Evolutionary origin and consequences of uniparental mitochondrial inheritance. AB - In the great majority of sexual organisms, cytoplasmic genomes such as the mitochondrial genome are inherited (almost) exclusively through only one, usually the maternal, parent. This rule probably evolved to minimize the potential spread of selfish cytoplasmic genomic mutations through a species. Maternal inheritance creates an asymmetry between the sexes from which several evolutionary consequences follow. Because natural selection on mitochondria operates only in females, mitochondrial mutations may have more deleterious effects in males than in females. Strictly uniparental inheritance creates asexual mitochondrial lineages that are vulnerable to mutation accumulation (Muller's ratchet). There is evidence that over evolutionary time mitochondrial genomes have indeed accumulated slightly deleterious mutations. Mutation accumulation in animal mitochondrial genomes is probably slowed down mainly by two processes: a severe reduction in germline mitochondrial genome copy number at some point in the life cycle, enabling more effective elimination of mutations by natural selection, and occasional recombination between maternal and paternal mitochondrial genomes following paternal leakage. PMID- 11041519 TI - Germline passage of mitochondria: quantitative considerations and possible embryological sequelae. AB - Using a semi-quantitative review of published electron micrographs, we have explored the passage of mitochondria from one generation to the next through the cytoplasm of the human female germ cell. We propose a testable hypothesis that the mitochondria of the germline are persistently 'haploid' (effectively carrying just one mitochondrial chromosome per organelle). For mitochondria, the passage through germ cell differentiation, oogenesis, follicle formation and loss could constitute a restriction/amplification/constraint event of a type previously demonstrated for asexual purification and refinement of a nonrecombining genome. At the restriction event (or 'bottleneck') in the human primordial germ cell, which differentiates in embryos after gastrulation, there appear to be <10 mitochondria per cell. From approximately 100 or so such cells, a population of > or =7 x 10(6) oogonia and primary oocytes is produced in the fetal ovaries during mid-gestation, with mitochondria numbering up to 10000 per cell, implying a massive amplification of the mitochondrial genome. A further 10-fold or greater increase in mitochondrial numbers per oocyte occurs during adult follicular growth and development, as resting primordial follicles develop to preovulatory maturity. So few are the numbers of oocytes that fertilize and successfully cleave to form an embryo of the new generation, that biologists have long suspected that a competitive constraint lies behind the generational completion of this genetic cycle. I propose that maintaining the integrity of mitochondrial inheritance is such a strong evolutionary imperative that features of ovarian follicular formation, function, and loss could be expected to have been primarily adapted to this special purpose. To extend the hypothesis further, the imperative of maintaining mitochondrial genomic integrity in a population could explain why women normally become sterile a number of years before there is depletion of ovarian follicles and endocrine ovarian failure (i.e. why there is 'an oopause' preceding the menopause). Plausible explanations might also follow for several well-known and puzzling reproductive difficulties, including recurrent miscarriage, unexplained infertility, and persistent failure of IVF embryos to cleave or to implant. Current experimental laboratory manoeuvres that might circumvent mitochondrial shortcomings (such as cytoplasmic transfusion and karyoplast exchange) are examined and possible clinical hazards identified. PMID- 11041520 TI - Mitochondrial morphology in human fetal and adult female germ cells. AB - The aim of this study has been to observe, by electron microscopy, the morphological changes affecting mitochondria and associated organelles in the human female germ cell during oogenesis, maturation and fertilization. In the primordial germ cell (PGC), rounded mitochondria with a pale matrix and small vesicular cristae are disposed near the nucleus and significantly increase in number during PGC migration and settlement in the gonadal ridge, where they differentiate into oogonia. In these early stages of mammalian oogenesis, aggregates of mitochondria are typically clustered around or in close relationship with the nuage. In oocytes at early prophase stage, mitochondria proliferate while aligned along the outer surface of the nuclear membrane, contain a more dense matrix than before, and have lamellar cristae. Oocytes of primordial and primary follicles mostly contain round or irregular mitochondria whose matrix has become very light. These mitochondria show typical parallel, arched cristae, and are clustered near the nucleus with other organelles forming the Balbiani's vitelline body. When follicles grow, the mitochondria of the oocytes become even more numerous and are dispersed in the ooplasm. Both paranuclear accumulation and subsequent dispersion of mitochondria in the cytoplasm are likely to be regulated by microtubules. By ovulation, mitochondria are the most prominent organelles in the ooplasm. They form voluminous aggregates with smooth endoplasmic reticulum (SER) tubules and vesicles. These mitochondrial SER aggregates (M-SER) and the mitochondrial-vesicle complexes (MV) could be involved in the production of a reservoir of substances or membranes anticipating subsequent fertilization and early embryogenesis. Just after fertilization, the mitochondria of the oocyte undergo a further substantial change in size, shape, and microtopography. In the pronuclear zygote, mitochondria concentrate around the pronuclei. During the first embryonic cleavage divisions, round or oval mitochondria with a dense matrix and few arched cristae are gradually replaced by elongated ones with a less dense matrix and numerous transverse cristae. A progressive reduction in size and number of M-SER aggregates and MV complexes also occurs. In summary, oocyte mitochondria show dynamic morphological changes as they increase in number and populate different cell domains within the oocyte. They form complex relationships with other cell organelles, according to the different energetic -metabolic needs of the cell during differentiation, maturation, and fertilization, and are ultimately inherited by the developing embryo, where they eventually assume a more typical somatic cell form. PMID- 11041521 TI - Mitochondrial morphology during preimplantational human embryogenesis. AB - The structure, distribution, and function of mitochondria during human oogenesis and early development is reported. Oogonia show a sparse and even distribution of mitochondria, which are oval or elongated. Except around nuclei, growing oocytes from small antral follicles have more dense rounded or oval mitochondria, associated with the rough endoplastic reticulum. Mitochondria in fully grown, germinal vesicle (GV) oocytes present an inert appearance, with a dense matrix and a few arch-like or transverse cristae. At this stage mitochondria are usually absent from the cortical part of the cytoplasm. Mitochondria in metaphase I and II oocytes, including fertilized oocytes, present a similar structure, but they are numerous and evenly spread in the ooplasm, associating closely with vesicles or aggregates of tubular smooth endoplasmic reticulum. The most substantial change in distribution occurs at the pronuclear stage, when there is a central conglomeration of mitochondria around the pronuclei in both monospermic and dispermic embryos, which persists up to syngamy. In structure and distribution, mitochondria in blastomeres of 2-16-cell embryos remain virtually unchanged and resemble those of mature oocytes, though perinuclear aggregation can be evident. Mitochondria are usually excluded from meiotic and mitotic spindles but locate peripherally, apparently providing energy for centrosomal, cytoskeletal, and chromosomal activity during cell division. Morphogenetic changes in mitochondrial structure occur in the 8-cell cleaving embryo, the morula and the blastocyst (apparently accompanying the onset of nuclear and mitochondrial transcription), when they become progressively less electron dense and often develop clear areas in their matrices. Elongating mitochondria with inner mitochondrial membranes arranged into transverse cristae appear in expanding blastocysts, in the trophoblast, embryoblast, and endodermal cells. These mitochondria seem to play a role in blastocyst differentiation, expansion, and hatching, with their morphological changes reflecting increased cellular activity. PMID- 11041522 TI - Chromosomal non-disjunction in human oocytes: is there a mitochondrial connection? AB - The frequency of chromosome abnormalities due to non-disjunction of maternal chromosomes during meiosis is a function of age, with a sharp increase in the slope of the trisomy-age curve between the ages of 30 and 40 years. The basis of this increase, which is a major cause of birth defects, is unknown at present. In recent years, mutations in mitochondrial (mt) DNA have been associated with a growing number of disorders, including those associated with spontaneous deletions of mtDNA (deltamt DNAs). Intriguingly, these pathogenic deltamtDNAs, which are present at extremely high levels in certain patients, are also present at extremely low levels (detectable only by polymerase chain reaction) in normal individuals. The proportion of such deltamtDNAs in normal muscle is a function of age; the shape of this curve is exponential, with the accelerating part of the curve beginning at approximately 30-40 years. We postulate that, as well as muscle and brain, a similar time-dependent accumulation of deltamtDNAs also occurs in normal oocytes. Since deltamtDNAs are functionally inactive, an accumulation of such aberrant genomes could eventually compromise ATP-dependent energy-utilization in these cells. Furthermore, these deficiencies would also affect the function of the somatic follicular cells that surround, and secrete important paracrine factors to, the oocyte. If there is indeed an age-associated relationship between deltamtDNAs and oocyte age, perhaps errors in meiosis (which is almost certainly an energy, and ATP, dependent process) are related to mutations in mtDNA (primarily deletions, but perhaps point mutations as well) in oocytes and/or the surrounding somatic cells, which result in deficiencies in both mitochondrial function in general and oxidative energy metabolism in particular. This hypothesis would explain many of the non-Mendelian features associated with maternal age-related trisomies, e.g. Down's syndrome. PMID- 11041523 TI - Intrafollicular influences on human oocyte developmental competence: perifollicular vascularity, oocyte metabolism and mitochondrial function. AB - While genetic and epigenetic factors have been associated with the developmental competence of human oocytes and embryos produced by IVF, how such factors develop or influence the oocyte remain to be explained. This paper reviews evidence which suggests that the degree of perifollicular vascular expansion associated with increased rates of blood flow are developmentally important for the generation of a normal follicle and competent oocyte. The degree of vascular development is follicle specific and differences between follicles might reflect their unique abilities to regulate angiogenic growth factor(s) production by the follicle cells in response to hypoxia. The notion that mitochondrial function in oocytes and early embryos could be influenced by intrafollicular conditions, and that differential patterns of mitochondrial segregation occur in blastomeres during early cleavage, is discussed with respect to the role of these organelles as critical determinants of developmental competence. PMID- 11041524 TI - Mitochondrial distribution and function in oocytes and early embryos. AB - Active mitochondria relocate during oocyte maturation or fertilization in several species. Detailed studies with hamster oocytes and early embryos reveal a pattern of active mitochondria migrating to surround the pronuclei to form a pattern that persists through the early cleavage stages. Although the functional significance of this relocation is unknown, it appears to be an important part of normal development in hamsters. Treatments that disrupt embryo development in vitro (such as the presence of inorganic phosphate or alteration of intracellular pH) also disrupt the normal pattern of mitochondrial distribution. Active mitochondria also reorganize during maturation in bovine oocytes and during fertilization in rhesus monkey oocytes. Examination of these changes in mitochondrial organization may provide insights into the regulation of normal embryo development and might serve as predictors of oocyte or embryo developmental competence. PMID- 11041525 TI - Toxic effects of oxygen on human embryo development. AB - The toxic effects of oxygen on the embryos of various animal species are reviewed. Methodologies for assessing embryonic damage are discussed and possible ways of preventing the damage are explored. Three methods of potentially minimizing oxidative damage to human embryos were tested using gametes, zygotes, and embryos from a clinical IVF programme: (i) decreasing the oxygen tension in the gas phase used for culture during insemination, fertilization, and embryo growth; (ii) changing the formulation of culture media to include some components designed to protect against oxidative damage; and (iii) reducing the duration of insemination to minimize the effect of oxidative damage caused by spermatozoal metabolism. Fertilization, cleavage, embryo utilization, pregnancy, and embryo implantation rates were used to monitor these changes. Although all three methods gave an increase in success rates, there was still a dramatic decrease in success with patient age. It is suggested that, although the system of handling and culturing embryos can be optimized with respect to embryonic mitochondrial function, there are inherent age-related defects in oocytes and embryos that are still more fundamental than the environmental conditions of the embryo. PMID- 11041526 TI - Spontaneous and artificial changes in human ooplasmic mitochondria. AB - Our research has focused on promoting the development of compromised embryos by transferring presumably normal ooplasm, including mitochondria, to oocytes during intracytoplasmic insemination. Because of the enigma of mitochondrial heteroplasmy, the mixing of populations of oocyte cytoplasm has provoked considerable debate. We are currently investigating oocyte mitochondrial (mt) DNA mutations and the effects of ooplasmic transplantation on mitochondrial inheritance and mitochondrial functionality. Ageing human oocytes could accumulate mtDNA deletions, which might lead to detrimental development. Elimination of abnormal, rearranged mtDNA, such that the offspring inherit only normal mitochondria, is postulated to occur by a mtDNA 'bottleneck'. Among compromised human oocytes (n = 74) and early embryos (n = 137), investigations have shown the occurrence of deltamtDNA4977, the so-called common deletion, to be 33% among oocytes and 8% among embryos. Using a nested polymerase chain reaction (PCR) strategy of long followed by short PCR, another 23 novel mtDNA rearrangements were found: various rearrangements were present in 51% of the oocytes (n = 295) and 32% of early embryos (n = 197). The difference in the percentage of mtDNA rearrangements between oocytes and embryos was significant (P < 0.0001) and implies that there could be a process of selection as fertilized oocytes become embryos. There was no significant relationship between the percentage of human oocytes or embryos that contained mtDNA rearrangements and age. The first series of ooplasmic transfers have been performed in women with repeated implantation failure associated with slow and morphologically abnormal development of their embryos. In a total of 23 attempts in 21 women, eight healthy babies have been born and other pregnancies are ongoing. By examining the donor and recipient blood samples it is possible to distinguish differences in their mtDNA fingerprint. A small proportion of donor mitochondrial DNA was detected in samples with the following frequencies: embryos (six out of 13), amniocytes (one out of four), placenta (two out of four), and fetal cord blood (two out of four). Ooplasmic transfer can thus result in sustained mtDNA heteroplasmy representing both the donor and recipient. PMID- 11041527 TI - Mitochondrial ultrastructure in embryos after implantation. AB - Information on the morphology of mitochondria during embryogenesis is scattered in the literature, but there appears to be a consistent pattern. During early organogenesis, the embryo is in a state of relative hypoxia associated with a major decrease in terminal electron transport system activity and a marked increase in anaerobic glycolysis. Ultrastructural studies of a 14-somite monkey embryo and day 10 and 12 rat embryos, together with a review of the literature, led us to determine that this hypoxic stage is characterized by vesiculation of the mitochondrial inner membranes, or cristae. Starting in the late morula stage and continuing during early postimplantation embryogenesis, the cristae increase but appear tubular or vesicular. After the end of neurulation, and with the onset of vascular perfusion of embryonic tissues, the cristae gradually become lamellated; by the limb bud stage they appear more mature. We suggest that new cristae derive from blebs of the inner mitochondrial membrane and that with maturation these blebs collapse, giving them a lamelliform appearance. The delamellated state of the cristae might inactivate oxidative phosphorylation to protect the embryo from toxic respiratory end-products that could accumulate in an embryo before there is vascular perfusion. Consistent with this hypothesis, mitochondrial diameters in the developing heart of monkey and rat embryos were approximately twice those found in skin and neural tube. PMID- 11041528 TI - Mitochondrial DNA segregation in the developing embryo. AB - Mitochondrial (mt)DNA is strictly maternally inherited in mammals; new mutations thus segregate along maternal lineages without the benefit of homologous recombination with mtDNA of paternal origin. Despite the high mtDNA copy number (approximately 100000 or more) in mature oocytes, and despite the relatively small number of cell divisions during oogenesis, mtDNA sequence variants segregate rapidly between generations. This paradoxical behaviour has been ascribed to the presence of a mtDNA 'bottleneck' in oogenesis or early embryogenesis. The nature and size of this bottleneck have been the subject of much controversy. This review argues that segregation of mtDNA sequence variants in the female germline occurs primarily during mitosis in the oocyte precursor population. Segregation is rapid because the precursor cells (primordial germ cells and oogonia) contain a relatively small number of mtDNA templates (the bottleneck) and because the replication of mtDNA is under relaxed control. For the most part, the process appears similar in mice segregating polymorphic sequence variants and in human pedigrees segregating pathogenic point mutations. In particular, there is no evidence for selection against high levels of pathogenic mtDNA point mutations in oogenesis, in early embryonic development, or in fetal development, thus suggesting that efficient respiratory chain function is not critical until post-natal life. These results have important practical implications for clinical genetics. PMID- 11041529 TI - Transmission of the human mitochondrial genome. AB - The segregation and transmission of mitochondrial genomes in humans are complicated processes, but are particularly important for understanding the inheritance and clinical abnormalities of mitochondrial disorders. This review describes three aspects of mitochondrial genetics. First, that the segregation and transmission of mitochondrial (mt)DNA molecules are likely to be determined by their physical association within the organelles and by the dynamics of mitochondrial structure and subcellular organization. Second, that the transmission of heteroplasmic mtDNA sequence changes from one generation to the next often involves rapid shifts in allele frequency. For >20 years, the standard explanation has been that there is a developmental bottleneck in which, at some stage of oogenesis, there is a reduction in the effective number of mitochondrial units of inheritance. The third aspect is that ongoing analyses of the segregation and transmission of pathogenic mtDNA mutations indicate the operation of multiple genetic processes. Thus, the segregation and transmission of mtDNA mutations occurs predominantly, but not exclusively, under conditions of random genetic drift. However, there is also evidence for bias due to incomplete ascertainment of pedigrees and for negative selection of pathogenic mutations in rapidly dividing somatic tissues such as the white blood cell population. PMID- 11041530 TI - Towards reliable prenatal diagnosis of mtDNA point mutations: studies of nt8993 mutations in oocytes, fetal tissues, children and adults. AB - Prenatal diagnosis of mitochondrial DNA (mtDNA) mutations is technically possible, but has only rarely been attempted. This is largely because of uncertainty about the effects of mtDNA heteroplasmy, the mtDNA bottleneck, random segregation or selection of mtDNA species, and difficulty in correlating a particular mtDNA mutant load with clinical outcome. We have investigated the feasibility of prenatal diagnosis for two common mtDNA mutations at nucleotide (nt)8993 by determining mtDNA mutant loads in human oocytes and by reviewing data on 56 pedigrees with these mutations, and by reviewing six studies on mtDNA mutations in human fetuses. Data from heteroplasmic human and mouse oocytes demonstrate that the bottleneck occurs in early oogenesis. Analysis of mutant loads of the nt8993 mutations in fetal and adult tissues confirms that there is no substantial tissue variation, implying that the mutant load in a prenatal sample will represent the mutant load in other fetal tissues. The two nucleotide 8993 mutations each show a strong correlation between mutant load and symptom severity and between maternal blood mutant load and risk of a severe outcome. We generated empirical data for calculating recurrence risk and predicting the clinical outcome of a given mutant load. These predictive data can be used (cautiously) for genetic counselling and prenatal diagnosis of nucleotide 8993 mutations. PMID- 11041531 TI - Antidepressant drug exposure is associated with mRNA levels of tyrosine receptor kinase B in major depressive disorder. AB - 1. Recent studies have provided support for the notion that the high affinity neurotrophin receptor tyrosine receptor kinase B (trk B) may be involved in the treatment of depression. 2. Using a quantitative RT-PCR approach trk B mRNA levels were determined in brain material from cerebellum, temporal cortex, and frontal cortex of control specimen and patients with major depressive disorder, schizophrenia and bipolar disorder (15 subjects each). 3. Interestingly, elevated trk B mRNA levels were found in cerebellum (3.6-fold) in patients with major depressive disorder, reaching statistical significance (p=0.03). 4. The major depressive disorder-on drugs group differed from controls (p=0.006) in the cerebellum. 5. Since only patients with major depressive disorder received antidepressants, elevated trk B mRNA levels are possibly related to drug treatment. PMID- 11041532 TI - The effects of antipsychotic medication on saccadic eye movement abnormalities in Huntington's disease. AB - 1. The study was a prospective open trial done to determine the effect of antipsychotic medication on saccadic eye movements in patients with Huntington's disease (HD). The authors tested 16 outpatients with HD, 8 of whom were drug free and 8 of whom were on antipsychotic medication. We also tested 24 healthy control subjects. 2. The antisaccade task was used to investigate the voluntary control of saccadic eye movements. Antisaccade latencies and error rates were used as outcome measures. Both HD patient groups showed significantly more antisaccade abnormalities than controls and the rate of abnormalities was very similar between the drug-free and medicated patient groups. 3. The results suggest that these abnormalities are related to underlying neuropathology rather than effects of medication. Either specific dysfunction of the dorsolateral prefrontal cortex or of the subcortical circuit connecting this cortical area to the superior colliculus via the striatum are likely to be responsible for the generation of such antisaccade eye movement abnormalities. PMID- 11041533 TI - Comparison of the effects of zolpidem and triazolam on nocturnal sleep and sleep latency in the morning: a cross-over study in healthy young volunteers. AB - 1. Zolpidem (ZPD, 10 mg) was directly compared with triazolam (TRZ, 0.25 mg), a benzodiazepine hypnotic of a short action comparable to ZPD. The compounds were given to healthy young subjects for three nights, in a crossover design. 2. Polysomnographic data of three 150-min sections of the nights as well as the whole nights were analyzed, to clearly detect the proper effects of the very short acting hypnotics, which might be missed in the analysis of whole night. 3. Time courses were significantly different between the two compounds in the ratios (%) of stage wake (SW), stage 2 (S2), slow wave sleep (SWS) and stage REM (SR). 4. Compared to the baseline, SWS was increased by ZPD on the first night, not by TRZ. The separate analysis of the three 150-min sections revealed an increase of SWS during the first 150-min of the ZPD night, suggesting a proper action of ZPD to augment SWS. An increase of S2 and a decrease of SR were caused by TRZ, not by ZPD. However, the separate analysis indicated that ZPD might reduce SR during the first 150-min, which was cancelled by a subsequent rebound increase in the whole night analysis. 5. During the withdrawal period, TRZ, not ZPD, increased SW and SR with worsening of mood in the morning. ZPD did not affect sleep latency in the morning, while TRZ caused a trend of the reduction. PMID- 11041534 TI - Risperidone versus clozapine in treatment-resistant schizophrenia: a randomized pilot study. AB - 1. The atypical antipsychotic risperidone may constitute an alternative to clozapine, the current treatment of choice for refractory schizophrenia. The objectives of this study were to evaluate the effectiveness of risperidone in comparison to clozapine in everyday practice and to assess the feasibility of a pragmatic trial procedure. 2. Patients were randomly assigned to open-label clozapine or risperidone treatment for 10 weeks and treatment outcomes were assessed blindly. Twenty-one patients were recruited and nineteen entered the randomized phase. 3. Five of 10 participants allocated to clozapine and one of nine risperidone participants dropped out before study completion. Five clozapine patients and six risperidone patients achieved clinical improvement, defined as a 20% decrease in the Positive and Negative Symptom Scale (PANSS) total score. No significant differences between the groups were detected in baseline or endpoint positive or negative symptoms, disease severity, or global or social functioning scores. Patients' opinion on the drugs did not differ between groups. 4. The findings of the intention-to-treat analysis of this study corroborates previous findings that risperidone may be equally effective as clozapine, and supports the feasibility and need of a multicenter randomized pragmatic trial with sufficient power to detect differences between treatments. PMID- 11041535 TI - Differential effects of MK-801 on cerebrocortical neuronal injury in C57BL/6J, NSA, and ICR mice. AB - 1. Antagonists of the N-methyl-D-aspartate (NMDA) glutamate (Glu) receptor, including [(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate], dizocilpine maleate (MK-801), injure pyramidal neurons in the posterior cingulate/retrosplenial (PC/RS) cortex when administered systemically to adult rats and mice. 2. These results have, to our knowledge, only been reported previously in Harlan Sprague Dawley albino rats and International Cancer Research (ICR) mice, an outbred albino strain. 3. Male Non-Swiss Albino (NSA) mice, an albino outbred strain, and male C57BL/6J (B6) mice, a pigmented inbred strain, were injected systemically with 1 mg/kg of MK-801 in the first experiment. This dose of MK-801 reliably produces cytoplasmic vacuoles in neurons in layers III and IV of the PC/RS cortex in 100% of ICR mice treated 4. There was a significant difference in the number of vacuolated neurons in B6 and NSA mice, as assessed by ANOVA. The NSA were not significantly different than previously examined ICR mice, but the B6 had fewer vacuolated neurons than either of the two outbred strains. 5. In the second experiment, male NSA, ICR, and B6 mice were injected systemically with a high dose, 10 mg/kg, of MK-801. This dose has been demonstrated to result in necrosis in the same population of neurons injured by lower doses of MK-801. 6. An ANOVA indicated that there was a significant difference among the three strains of mice, and a Fisher's protected t revealed that the B6 mice were significantly different from both the NSA and ICR, but that, with our test, those two strains were indistinguishable. 7. Male ICR, NSA, and B6 mice were tested in the holeboard food search task 5 hours after 1 mg/kg of MK-801. There were significant differences between the strains in performance both pre and posttreatment. The effect of the drug was not statistically significant. 8. These results suggest that there may be a genetically mediated difference in the reaction to NMDA receptor antagonists, a finding which may be important given the NMDA receptor hypofunction hypothesis for the etiology of schizophrenic symptoms. PMID- 11041536 TI - Neurotoxic effects of caulerpenyne. AB - 1. In this paper the authors tested the effect of caulerpenyne (CYN), a sesquiterpene synthesized by the green alga Caulerpa taxifolia onto the central nervous system of the leech Hirudo medicinalis. Investigations have been performed with three different approaches: neuroethological, electrophysiological and neurochemical techniques. 2. CYN application mimics the effect of a nociceptive stimulation (brushing), eliciting a clear-cut potentiation of the animal swim response to the test stimulus (non associative learning process such as sensitization). This effect is similar to that one induced by the endogenous neurotransmitter serotonin (5HT). 3. CYN strongly reduces the after hyperpolarization (AHP) recorded from T sensory neurons. This effect overlaps that one produced by 5HT, but it is not affected by the serotonergic antagonist methysergide. 4. The decrease of AHP amplitude due to CYN application is observed also in presence of apamin, a blocking agent of Ca++-dependent K+ channels, suggesting that CYN is acting through the inhibition of the Na+/K+ electrogenic pump. 5. The depression of the AHP driven by CYN is not prevented by application of MDL 12330A, an adenylate cyclase inhibitor. On the other hand MDL 12330A counteracts the reduction of AHP due to 5HT application. 6. Incubation of the leech central nervous system with CYN induces the phosphorylation of proteins of 29, 50, 66 and 100 kDa. This pattern of phosphorylation is similar to that one elicited by 5HT treatment. 7. The data demonstrate that CYN exerts remarkable effects on leech neurons by acting onto specific molecular targets such as the Na+/K+ ATPase. This effect may influence important neural integrative functions and may explain the sensitizing action produced by the toxin on swim induction. Finally, caulerpenyne does not act through the pathways involved in the 5HT action, and its effect is not mediated by the second messenger cyclic AMP. The mechanism of action of CYN are still under investigations. PMID- 11041537 TI - Differential behavioural and neurochemical effects of para-methoxyamphetamine and 3,4-methylenedioxymethamphetamine in the rat. AB - 1. This study was prompted by recent deaths that have occurred after recreational administration of the substituted amphetamine para-methoxyamphetamine (PMA). Because relatively little is known regarding its mechanism(s) of action, its effects on physiological, behavioural and neurochemical parameters were compared with the well known effects of 3,4-methylenedioxymethamphetamine (MDMA). 2. Equivalent doses of PMA (5-20 mg/kg) produced greater hypothermia than MDMA at an ambient temperature of 20 degrees C. At 30 degrees C, PMA continued to evoke hypothermia except the highest dose where hyperthermia ensued. MDMA altered body temperature only at the highest dose where hyperthermia also resulted. 3. At both 20 and 30 degrees C, MDMA stimulated locomotor activity whereas PMA had modest effects and then, only at high doses. 4. In vivo chronoamperometry was used to measure the effect of MDMA and PMA on release, and inhibition of uptake, of serotonin (5-HT) and dopamine (DA) in the dorsal striatum of anaesthetised rats. As expected, MDMA evoked release of DA and inhibited uptake of both DA and 5-HT. By contrast, PMA was a relatively weak releasing agent and did not inhibit DA uptake. However, PMA potently inhibited uptake of 5-HT. 5. Taken together these data suggest that the acute adverse effects of PMA are more likely to be associated with alterations in serotonergic rather than dopaminergic neurotransmission. PMID- 11041538 TI - Characterization of benzodiazepine receptors in the cerebellum. AB - 1. The goals of the work reported here were to further characterize benzodiazepine/GABA(A) (BDZR) receptor heterogeneity in the cerebellum and to measure the affinities and selectivities of structurally diverse benzodiazepines at each site identified. 2. Five chemical families were included in these studies. These were 1,4-benzodiazepines (flunitrazepam), imidazobenzodiazepines (RO15-1788 and RO15-4513 and RO16-6028), beta-carbolines (Abecarnil) and pyrazoloquinolines (CGS 8216, CGS 9895 and CGS 9896). 3. Saturation and competition binding assays were combined with powerful data analysis software developed in our laboratory. Among the capabilities of this software is the identification of multiple binding sites for a cold ligand using a non-selective labeled ligand that binds with equal, but high, affinity to all the binding sites 4. Saturation binding assays using either [3H]-RO15-1788 or [3H]-RO15-4513 revealed only one apparent binding site, with a higher affinity for RO15-4513 than for RO15-1788. However, using [3H]-RO15-4513 for the competition binding studies in the cerebellum, together with our data analysis software, led to the identification of two distinct binding sites with equal densities for the diverse benzodiazepines studied. 5. In rat cerebellum one of the sites identified corresponds to GABA(A) receptors exhibiting alpha1 subunit pharmacology and the other to GABA(A) receptors exhibiting alpha6 subunit pharmacology. In general, the diverse families of BDZR ligands studied had much lower affinities for the alpha6 containing receptors. PMID- 11041540 TI - Modulation of kainate--activated currents by diazoxide and cyclothiazide analogues (IDRA) in cerebellar granule neurons. AB - 1. Patch-clamp technique was used in primary cultures of cerebellar granule neurons to study the modulation of the cyclothiazide analogue (IDRA21) and of the diazoxide derivative (IDRA 5) on KA-evoked currents. 2. The dose-response of kainic acid (KA) reveals an EC50=90 microM and an Hill coefficient of 1.3. IDRA 21 and cyclothiazide potentiate KA-evoked current in a dose dependent way, being cyclothiazide more potent but less efficacious than IDRA 21. Conversely IDRA 5 acts as a negative modulator of KA evoked -current. 3. Application of IDRA 21 and cyclothiazide results in a current potentiation of 125+/-18% and 80+/-12% respectively, while IDRA 5 decreases KA-current (-21+/-5%). Coapplication of cyclothiazide and IDRA 21 produces a potentiation of 110+/-17%, suggesting a competition of the two drugs for the same site. 4. In the same experimental model we studied the ability of IDRA compounds of promoting toxicity through AMPA receptor activation. Under basal conditions AMPA treatment (50 microM for 1 hour) results in a negligible excitotoxicity. 5. In contrast similar treatment with AMPA + IDRA 21 (1 mM) or + IDRA 5 (1 mM) or + cyclothiazide (100 microM) induces citotoxicity. The neurotoxic damage induced by IDRA 21 and cyclothiazide is blocked by GYKI 53655 (50 microM) and by NBQX (10 microM). Interestingly GYKI and NBQX are ineffective in reducing IDRA 5 toxicity. PMID- 11041539 TI - Different mechanisms for dopaminergic excitation induced by opiates and cannabinoids in the rat midbrain. AB - 1. The mechanism underlying morphine and cannabinoid-induced excitation of meso accumbens and nigro-striatal dopaminergic neurons was investigated by extracellular single unit recording techniques coupled with antidromic activation from the nucleus accumbens and striatum respectively, in unanesthetized rats. 2. The intravenous administration of cumulative doses (1-4 mg/kg) of morphine, dose dependently increased the firing rate of dopaminergic neurons projecting to the nucleus accumbens and neostriatum, while the same doses inhibited the activity of pars reticulata neurons of the substantia nigra. Both effects were antagonized by naloxone (0.1 mg/kg i.v.) but not by the selective CB1 receptor antagonist SR 141716A (1 mg/kg i.v.). 3. The intravenous administration of cumulative doses (0.125-0.5 mg/kg) of delta9-tetrahydrocannabinol (delta9-THC) also increased the firing rate of meso-accumbens and nigro-striatal dopaminergic neurons; this effect was antagonized by SR 141716A (1 mg/kg i.v.), but not by naloxone. 4. Furthermore, nor delta9-THC up to a dose of 1 mg/kg, maximally effective in stimulating dopamine neurons, neither SR 141716A (1 mg/kg i.v.) at a dose able to reverse the stimulatory effect of delta9, THC on dopamine cells, did alter the activity of SNr neurons. 5. The results indicate that morphine and delta9-THC activate dopaminergic neurons through distinct receptor-mediated mechanisms; morphine may act by removing the inhibitory input from substantia nigra pars reticulata neurons (an effect mediated by mu-opioid receptors). Alternatively, the delta9-THC-induced excitation of dopaminergic neurons seems to be mediated by CB1 cannabinoid receptors, while neither mu-opioid receptors nor substantia nigra pars reticulata neurons are involved. PMID- 11041541 TI - Neonatal treatment with L-name (NG-nitro-L-arginine methyl ester) attenuates stereotyped behavior induced by acute methamphetamine but not development of behavioral sensitization to methamphetamine. AB - 1. The neurodevelopmental hypothesis of schizophrenia postulates that disturbed nitric oxide (NO) function during neuronal development is one of premorbid factors for schizophrenia in later life. 2. The aim of present study is to investigate behaviorally whether neonatal inhibition of nitric oxide synthase (NOS) affects dopaminergic function, the abnormality of which may be ascribed to a major pathophysiology of schizophrenia. 3. Male rat pups were injected daily with NOS inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), from postnatal day (PD) 1 to 14. 4. When methamphetamine (MAP) was challenged on PD42, MAP-induced stereotypy was significantly attenuated in the L-NAME treated rats. The development of sensitization to the stereotypy-inducing effect of MAP, however, was not prevented with neonatal L-NAME. 5. These results suggest that decreased NO production during neonatal period may disturb normal maturation of dopaminergic system and result in impaired dopaminergic function in adult period. PMID- 11041542 TI - Evaluation of behavioral disinhibition in P/NP and HAD1/LAD1 rats. AB - 1. Two lines of rats specifically bred for alcohol preference were exposed to two different behavioral tasks that required behavioral inhibition to successfully solve. 2. Learning and performance of a step-down passive avoidance task and a differential reinforcement of low-rate responding task were studied in P/NP and HAD1/LAD1 rats. 3. While the P rats had difficulty in learning both tasks, HAD1, LAD1 and NP rats performed at control levels. 4. These data suggest that P rats, but not HAD1 rats, may have problems learning tasks that require inhibition of ongoing or previously learned behaviors. PMID- 11041543 TI - Risperidone, neuroleptic malignant syndrome and probable dementia with Lewy bodies. AB - 1. Conflicting reports are available regarding the sensitivity of patients with Dementia with Lewy bodies (DLB) to risperidone. 2. The authors studied a rare familial case of probable DLB, who developed a documented episode of neuroleptic malignant syndrome (NMS) following the exposure to risperidone. Previously, the patient had had an episode of NMS on trifluoperazine. 3. The discontinuance of risperidone, in combination with a mild increase of dopaminergic therapy, led to a complete recovery in few days. 4. In patients with DLB, a continued vigilance for extrapyramidal side effects, including NMS, would be advisable during the use of risperidone. PMID- 11041544 TI - Transepithelial chloride conductance in amphibian skin: regulatory mechanisms and localization. AB - The transepithelial transport of Na+ by amphibian skin must be accompanied by the corresponding anion, Cl-, and much effort has been devoted to the characterization of Cl- transport. The transepithelial Cl- conductance, G(Cl), is activated by voltage and adenosine 3',5'-cyclic monophosphate (cAMP), shows rectification, requires the presence of Cl- in the pathway and is influenced by factors modifying intracellular signalling cascades and by metabolic poisons such as cyanide (CN-). Until recently, these findings were interpreted as strong evidence for a transcellular path, for which, given the impermeability of the principal cells for Cl-, the mitochondria-rich cells (MRC) are the only candidate. This was supported by the apparent parallelism between G(Cl) and the density of MRC (D(mrc)). Data accumulated in recent years, however, raise serious doubts as to the validity of this concept. The single-channel conductance derived from various techniques is too small by an order of magnitude to account for the observed G(Cl), the very slow time course of conductance activation is not reconcilable with any known membrane channel gating processes, a more thorough examination of the relationship between G(Cl) and D(mrc) fails to show any consistent pattern and analysis of current density immediately above the transporting epithelium using the vibrating voltage probe shows current peaks associated with only a small fraction of MRC, and even so, these current peaks account for about 20% of the transepithelial current. The remaining 80% of the current cannot be localized to specific structures. Given the increasing evidence for close cellular control of tight-junction function, the foregoing findings are equally consistent with an additional, major, paracellular pathway for Cl-. A comprehensive description of Cl- transport must await the final resolution of the transport pathway(s). PMID- 11041545 TI - Activation of the human, intermediate-conductance, Ca2+-activated K+ channel by methylxanthines. AB - This study demonstrated that the methylxanthines, theophylline, IBMX and caffeine, activate the human, intermediate-conductance, Ca2+-activated K+ channel (hIK) stably expressed in HEK-293 cells. Whole-cell voltage-clamp experiments showed that the hIK current increased reversibly and voltage independently after the addition of methylxanthines. In current-clamp experiments, theophylline dose dependently hyperpolarised the cell membrane from a resting potential of -18 mV to -56 mV. The methylxanthines did not affect large-conductance (BK) or small conductance (SK2), Ca2+-activated K+ channels, demonstrating that the effects were not secondary to a rise in intracellular Ca2+. However, the activation of hIK by theophylline required an intracellular [Ca2+] above 30 nM. The hIK current was insensitive to 8-bromoadenosine cyclic 3',5'-monophosphate (8-bromo-cAMP), forskolin, 8-bromoguanosine cyclic 3',5'-monophosphate (8-bromo-cGMP) and sodium nitroprusside. Moreover, in the presence of inhibitors of protein kinase A (PKA) or protein kinase G (PKG) theophylline still activated the current. Finally, mutation of the putative PKA/PKG consensus phosphorylation site (Ser334) had no effect on the theophylline-induced activation of hIK. Since the observed activation is independent of changes in PKA/PKG-phosphorylation and of fluctuations in intracellular Ca2+, we suggest that the methylxanthines interact directly with the hIK protein. PMID- 11041546 TI - Transport of magnesium by two isoforms of the Na+-Ca2+ exchanger expressed in CCL39 fibroblasts. AB - Cytoplasmic concentrations of Ca2+ ([Ca2+]i) and Mg2+ ([Mg2+]i) were measured with fluorescent indicators in CCL39 cells, a cell line established from Chinese hamster lung fibroblasts, transfected with complementary deoxyribonucleic acid (cDNA) of the Na+-Ca2+ exchanger isolated either from canine heart (NCX1) or from rat brain (NCX3). Raising extracellular [Mg2+] to 10 mM increased Mg2+ influx and the resultant change in [Mg2+]i (delta[Mg2+]i) was monitored with furaptra under Ca2+-free conditions. In control (vector-transfected) cells, delta[Mg2+]i at 45 min was similar with or without extracellular Na+ (130 mM or 0 mM) and when [Na+]i was raised by 1 mM ouabain treatment. delta[Mg2+]i in NCX1-transfected cells was attenuated significantly in the presence of 130 mM Na+, but became comparable to (or slightly larger than) that in control cells on either removal of extracellular Na+ or treatment with 1 mM ouabain. Cells expressing NCX3 showed an intermediate dependence of delta[Mg2+]i on Na+, probably reflecting a lower degree of expression of the exchanger protein. Extracellular Na+-dependent changes in [Ca2+]i (measured with fura-2 in the presence of extracellular Ca2+ and 10 microM ionomycin, a Ca2+ ionophore) were minimal in control cells, marked in the NCX1-transfected cells and intermediate in the NCX3-transfected cells. These results suggest that the Na+-Ca2+ exchanger (either NCX1 or NCX3) can transport Mg2+ and may play a role in the extrusion of magnesium from cells. PMID- 11041547 TI - Ca2+ and electrolyte mobilization following agonist application to the pancreatic beta cell line HIT. AB - We have investigated intracellular Ca2+ mobilization in oscillations of cytoplasmic Ca2+ in response to glucagon-like peptide 1 (GLP-1) and glucose in clonal HIT insulinoma cells with a confocal laser-scanning microscope (CLSM). We also used electron probe X-ray microanalysis to determine the GLP-1- and glucose induced changes in electrolyte levels in the cytoplasm and insulin granules of the cells. GLP-1 produced 10- to 35-s duration oscillations in cytoplasmic Ca2+ concentration ([Ca2+]i), both with and without Ca2+ in the extracellular solution, suggesting that Ca2+ is mobilized from intracellular Ca2+ stores, namely secretory granules. Glucose caused 1- to 3-min duration oscillatory increases in [Ca2+]i when the extracellular solution contained Ca2+. When the cells were cultured without Ca2+ (no Ca2+ added, 1 mM EGTA), an oscillatory [Ca2+]i increase of amplitude and short duration (12-35 s) was produced by 11 mM glucose, and the oscillation was inhibited by ruthenium red. X-ray microanalysis showed that stimulation with glucose increased the total Ca concentration in the cytoplasm and decreased it in the insulin granules with and without Ca2+ in the extracellular solution. The application of glucose significantly decreased K, and increased Na and C1 in the cytoplasm when the extracellular solution contained Ca2+. Our result also suggests that the [Ca2+]i oscillation induced by glucose is involved in the release of Ca2+ from intracellular Ca2+ stores through the ryanodine receptor, which is blocked by ruthenium red, and/or through the inositol trisphosphate receptor that may be present in the membrane of insulin granules. PMID- 11041548 TI - The kinetics of gap junction currents are sensitive to the ionic composition of the pipette solution. AB - Myocytes were isolated from neonatal rat hearts using an enzymatic procedure. Cell pairs were used to control the junctional voltage, V(j), and to measure the transjunctional current, I(j), using the dual voltage-clamp method. V(j) gradients provoked I(j) signals with voltage-dependent inactivation. During voltage pulses, I(j) remained virtually constant at ?V(j)? <40 mV. At ?V(j)?>40 mV, it inactivated with time to a residual level. The inactivation followed a single exponential. The time constant of I(j) inactivation, taui, and the size of I(j) at steady state, I(j,ss), were both sensitive to the ions in the pipette solution. I(j,ss) was smaller in the presence of tetraethylammonium aspartate (TEA+ aspartate-) than KC1, while taui was smaller in the presence of KC1 than TEA+ aspartate-. The modification of I(j,ss) is readily explained by a change in the residual conductance of the gap junction channels, gammaj,residual x The alterations in taui are correlated with a change in beta, the rate constant that describes the transition of the channel from the main state to the residual state. Pipette solutions may affect the kinetics of gap junction currents by altering the conductive and/or kinetic parameters. Computer simulations revealed a substantial influence of the latter, but only a marginal effect of the former. Conceivably, ions of the pipette solution may affect the kinetics of gap junction channels by screening surface charges of the channel wall. PMID- 11041549 TI - Calorimetry and respirometry in guinea pigs in hydrox and heliox at 10-60 atm. AB - We used direct calorimetry and respirometry to measure the total rate of heat loss (Qsigma) and of oxygen consumption (VO2) in guinea pigs in 1-atm (0.1 MPa) air and at 10-60 atm in either heliox (98% He, 2% O2) or hydrox (98% H2, 2% O2). Our objective was to determine if the physiological responses to these two gas mixtures were different and, if so, whether the differences were attributable to the thermal characteristics of the gases alone or were confounded by additional mechanisms. At 10-40 atm, Qsigma and VO2 were not significantly different in the two gas mixtures, whereas at 60 atm, Qsigma and VO2 were significantly higher in heliox than in hydrox. The VO2/Qsigma ratio suggested that the animals were not in thermal equilibrium in hyperbaria. Based solely on the differing thermal properties of the gas mixtures, a mathematical model predicted a Qsigma that was higher in hydrox than in heliox at all pressures. Two plausible explanations are suggested: one is an adaptive lowering of the surface temperature as a physiological response of the animal to the thermally more stressful hydrox environment, and the other is related to the narcotic suppression of the animal's activity by hydrox. PMID- 11041550 TI - Functional incorporation of exogenous proteins into the Xenopus oocyte membrane does not depend on intracellular calcium increase. AB - Fusion of membranes occurs in diverse biological events and, in most cases, Ca2+ greatly augments its rate. The aim of this work was to study the role played by Ca2+ when transplanting exogenous proteins into Xenopus oocyte membranes. Lipid vesicles carrying nicotinic acetylcholine (ACh) receptors (nAChRs) from Torpedo electroplaques were injected into oocytes. The time course of nAChR incorporation was assessed by recording ACh-evoked currents at different times from injection. An incorporation peak was found at 16 h, but responses were maintained for over 48 h. To assess the role played by Ca2+, two groups were considered: control and chelator-loaded oocytes. In the latter group, cells were incubated with 50 microM 1,2-bis (2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, tetrakis(acetoxymethyl)ester (BAPTA-AM) or loaded with ca. 5 nmol ethyleneglycol bis (beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) 2 h before nAChR injection. Both groups responded to ACh, although the current amplitude was smaller in chelator-loaded than in control cells. These results indicate that the slow fusion of lipoproteosome vesicles with the oocyte plasma membrane does not depend on intracellular Ca2+ increase and therefore belongs to the type called "constitutive". This membrane fusion process is thus different from those involved in resealing of disrupted oocyte membranes or in the fusion of cortical granules with the egg membrane. PMID- 11041551 TI - Characterization of whole-cell currents elicited by mechanical stimulation of Xenopus oocytes. AB - Whole-cell mechanosensitive current (I(ms)) in Xenopus oocytes was studied using the two-electrode voltage-clamp technique. I(ms) was evoked by mechanically pressing the oocyte surface with a glass micropipette. The current was found to depend on the amplitude of the stimulus, showed a time-dependent decay, and turned off immediately after the stimulus was removed. The current-voltage relationship for the peak current exhibited inward and outward rectification at negative and positive potentials, respectively, while that for the sustained current exhibited only inward rectification. I(ms) was significantly suppressed by 30 microM Gd3+. One millimolar amiloride also significantly suppressed the inward I(ms) at negative potentials, but not the outward one at positive potentials. Replacing extracellular Na+ with K+ did not change the current voltage relationship, whereas replacing extracellular Na+ with choline+ or tetraethylammonium+ significantly decreased the inward I(ms). The outward rectifier at positive potentials was abolished by replacing extracellular Cl- with gluconate-, by intracellular injection of 1,2-bis (2-aminophenoxy) ethane N,N,N',N'-tetraacetic acid (BAPTA), by extracellular application of anthracene-9 carboxylic acid, and by replacing extracellular Ca2+ with Mg2+. These results suggest that mechanical stimulation activates stretch-activated cation channels and Ca2+-activated Cl- channels, the latter being secondarily activated by an increase in intracellular Ca2+ concentration by Ca2+ influx through stretch activated cation channels. PMID- 11041552 TI - Modulation of corneal endothelial hydration control mechanisms by Rolipram. AB - Corneal stromal hydration is maintained by an active HCO3- transport mechanism located in the corneal endothelium. Whilst modulation of transport activity by changes in intracellular cAMP concentration have been noted, the site of effect is undefined. To resolve this question, the effects of Rolipram, a cAMP phosphodiesterase inhibitor, on endothelial physiology were determined. Addition of 0.1 mM Rolipram caused a threefold increase in intracellular cAMP with no change in cGMP. Associated with the increase in cAMP was a transient whole corneal thinning and a similarly transient increase in trans-endothelial potential difference, short-circuit current and resistance. The membrane potential hyperpolarized and the intracellular Na+ concentration decreased. The decreased intracellular Na+ was associated with an increased rate of Na+ extrusion between the endothelial cell and extracellular space. It is concluded that Rolipram increases the concentration of cAMP which activates the basolateral membrane Na+/K+-ATPase activity and increases net HCO3- transport. In addition there is a reduction in endothelial permeability which combined with the increase in pump activity may jointly explain the observed stromal thinning. The duplicity of responses indicates that if cAMP has a physiological role in regulating corneal hydration then it may operate on both the endothelial pump and the endothelial permeability. PMID- 11041553 TI - The cultured human gastric cells HGT-1 express the principal transporters involved in acid secretion. AB - HGT-1 is a human cell line sharing a number of physiological features with gastric parietal cells. HGT-1 cell monolayers were able to secrete H+ when stimulated with histamine (calculated external pH variation, deltapH(e) 0.46+/ 0.05) as assessed using the impermeant, pH-sensitive fluorescence dye 8 hydroxypyrene-1,3,6-trisulphonic acid, trisodium salt (HPTS). Treatment with 100 microM omeprazole inhibited the histamine-induced apical acidification by about 60%. Intracellular pH (pH(i)) measurements using the fluorescent pH-sensitive dye 2',7'-bis-carboxyethyl-5(6)-carboxyfluorescein (BCECF) demonstrated the expression of a functional, omeprazole-sensitive H+/K+-pump. A monoclonal antibody directed against the alpha subunit of the H+/K+-ATPase immunoprecipitated a 95-kDa protein from HGT-1 cells and human stomach which corresponds to the expected molecular size of the native protein. HGT-1 cells were also positive for the anion exchanger AE2 that is expressed in gastric parietal cells. In addition, we identified a histamine- and pH(i)-sensitive Na+/H+ exchanger in HGT-1 cells, which might correspond to the functional expression of the NHE4 isoform that has been detected in gastric epithelial cells as well as in primary cultured parietal cells. HGT-1 cells therefore display the principal features of parietal cells and might represent an interesting cell culture model for studying the regulatory mechanisms involved in acid secretion. PMID- 11041554 TI - Control of cell proliferation by cell volume alterations in rat C6 glioma cells. AB - K+ and Cl- channels are involved in regulating the proliferation of a number of cell types. Two main hypotheses have been proposed to explain the mechanism by which these channels influence cell proliferation: regulation of membrane potential and regulation of cell volume. In order to test these hypotheses, we measured, under different experimental conditions, the volume, membrane potential and rate of proliferation of C6 glioma cells. Cells cultured in control medium for 1-4 days were compared with cells cultured for the same period of time in the presence of broad spectrum channel blockers: tetraethylammonium, 5-nitro-2-(3 phenylpropylamino) benzoic acid (NPPB) and Cs+, in hypertonic media (29% increased osmolarity with NaCl, KCl or sucrose), in hypotonic medium (23% decreased osmolarity with H2O) or in the presence of the specific channel blockers, i.e. mast cell degranulating peptide, charybdotoxin or chlorotoxin. In all of these conditions, we observed a close correspondence between the rate of proliferation and the mean cell volume. The proliferation decreased when volume increased. Moreover, whereas control cells were flattened, spindle-shaped, bipolar or multipolar, cells cultured in media supplemented with NPPB, KCl or CsCl were round with few processes. Of the agents tested, only KCl and Cs+ depolarized the cells. These results show that alterations of the rate of proliferation by K+ and Cl- channel blockers or anisotonia are closely related with changes in cell volume or form but are not correlated with changes in membrane potential. PMID- 11041555 TI - Role of potassium channels in the control of renin secretion from isolated perfused rat kidneys. AB - This study aimed to assess the relevance of specific potassium channels, such as inwardly or outwardly rectifying and calcium-regulated potassium channels, to the control of renin secretion. For this purpose we examined the effects of the K+ channel blockers 4-aminopyridine (1 mmol/l), barium (100 micromol/l), tetraethylammonium (2 mmol/l) and apamin (200 nmol/l) on basal renin secretion, on renin secretion stimulated by isoproterenol (10 nmol/l) and on the inhibition of renin secretion by angiotensin II (100-300 pmol/l) in the isolated rat kidney perfused at constant pressure. Whilst all four K+ channel blockers increased renal vascular resistance, only 4-aminopyridine and barium attenuated isoproterenol-stimulated renin secretion in an additive fashion and augmented the inhibitory effect of angiotensin II. These effects of K+ channel blockers were not changed by the L-type calcium channel blocker amlodipine (5 pmol/l), indicating that their effects on renin secretion are not due to voltage-operated calcium influx. Our data, moreover, suggest that potassium efflux from renal juxtaglomerular cells is not important for the inhibitory action of angiotensin II on renin secretion. As a consequence it appears that the membrane potential of renal juxtaglomerular cells per se is relevant to renin secretion such that membrane depolarization inhibits the exocytosis of renin whilst hyperpolarization favors renin secretion. By their activity, potassium channels can contribute to membrane hyperpolarization and thus facilitate renin secretion. PMID- 11041556 TI - Elementary receptor currents elicited by a single pheromone molecule exhibit quantal composition. AB - Responses to single pheromone molecules, elementary receptor currents (ERC), recorded from antennal olfactory receptor neurones of the moth Bombyx mori were studied in situ, at the opened sensilla, using the voltage-clamp technique in transepithelial recordings. The amplitude parameters of the ERCs eliciting one action potential in the bombykol and bombykal receptor neurones were analysed. The ERC amplitude varied largely within a range of 0.5-5.5 pA. The amplitude histograms of the ERCs exhibited a series of peaks and shoulders. Quantal analysis of the amplitude distributions based on spectral methods revealed that a quantum event underlies the ERCs. Deconvolution of the ERC amplitude distributions using the distribution of baseline noise indicates that the ERCs exhibit discrete amplitude levels. The weighted average interval between the levels indicates that the quantal size is 0.6-0.7 pA. PMID- 11041557 TI - Inhibition of Jurkat-T-lymphocyte Na+/H+-exchanger by CD95(Fas/Apo-1)-receptor stimulation. AB - Mitogenic factors are known to stimulate the Na+/H+-exchanger (NHE), leading to cytosolic alkalinization and/or cell swelling. Conversely, a hallmark of apoptosis is cell shrinkage and CD95-induced apoptosis has been reported to be paralleled by cytosolic acidification. To assess whether the CD95-receptor regulates NHE activity in Jurkat T-lymphocytes, we performed conventional BCECF fluorescence measurements and SNARF flow cytometric analysis (FACS). The recoveries from acidifications following application of butyrate or a NH3 pulse were both abolished by a specific NHE-inhibitor, HOE694, indicating that they fully depend on NHE activity. Thus they were taken as a measure of NHE activity. CD95-receptor stimulation caused a cytosolic acidification and blunted the recovery from acidification following application of butyrate or a NH3 pulse. Moreover, the NHE-dependent alkalinization following osmotic cell shrinkage was almost abolished by CD95-receptor stimulation. As apparent from the effect of osmotic cell shrinkage, inhibition of the NHE by CD95-receptor stimulation was absent in Lck56-deficient J-CaM1.6 cells and restored by retransfection of J CaM1.6-cells with Lck56. CD95-receptor stimulation led within 4 h to a decrease of cellular ATP which could contribute to NHE inhibition. Treatment of Jurkat cells with the NHE inhibitor HOE694 accelerated CD95-induced DNA fragmentation. In conclusion, CD95-receptor stimulation inhibits NHE activity through a mechanism that depends directly or indirectly on the activation of the Src-like kinase Lck56. This effect contributes to CD95-induced cytosolic acidification, DNA fragmentation and cell shrinkage. PMID- 11041558 TI - Incubation in tissue culture media allows isolated rabbit proximal tubules to regain in-vivo-like transport function: response of HCO3-absorption to norepinephrine. AB - Using a new stop-flow perfusion technique with microspectrofluorometric determination of luminal fluid pH, we have studied which substrates or incubation conditions allow isolated rabbit proximal tubules to attain in-vivo-like rates of HCO3- absorption (J(HCO3)) and maximal responses of J(HCO3) to norepinephrine (NE). Essentially three incubation media were tested: plasma-like HCO(3-)-Ringer solution containing 5 mmol/l D-glucose (G-Ringer sol.), the same solution also containing 10 mmol/l lactate and 5 mmol/l L-alanine, (LAG-Ringer sol.), and two tissue culture media (DMEM and RPMI 1640). Compared to G-Ringer sol., application of LAG-Ringer sol. in the bath and/or lumen, or application of DMEM or RPMI 1640 in the bath either slightly increased or decreased J(HCO3) with borderline significance. However, RPMI 1640 plus 1 mmol/l pyruvate stimulated J(HCO3) by 55%. While NE (10(-5) mol/l), if applied in G-Ringer sol., had no effect, in the presence of LAG-Ringer sol. it increased J(HCO3) by approximately =40%, and in the presence of DMEM or RPMI 1640 it increased J(HCO3) by approximately =100%. This stimulation by NE followed Michaelis-Menten kinetics with an EC50 value of 0.25 micromol/l and was probably mediated by alpha1-adrenergic receptors. Additional cell pH measurements suggest that NE stimulates the basolateral Na+ HCO3- cotransporter which then becomes susceptible to inhibition by cAMP. We conclude that incubation in tissue culture media allows isolated proximal tubules to maintain a better functional state than the commonly used solutions with unphysiologically high substrate concentrations. PMID- 11041559 TI - Skeletal muscle HSP72 level during endurance training: influence of peripheral arterial insufficiency. AB - Heat shock protein 72 (HSP72), the inducible isoform of the HSP70 family, is synthesized in exercised rat muscles and in the ischaemic heart. To determine the isolated and combined effects of chronic ischaemia and repeated exercise on skeletal muscle HSP72 expression, male Wistar rats were subjected to unilateral occlusion of the iliac artery. Beginning 1 week after ischaemia, half the rats were exercised on a motor-driven treadmill once a day, 5 days/week, the other half were restricted to cage activity. Rats were sacrificed after 2, 4, or 8 weeks of endurance training, together with the age-matched sedentary rats. Tissue samples were obtained from the plantaris and the red portion of the quadriceps of both hind-limbs. Endurance-trained rats displayed significantly increased HSP72 levels in skeletal muscles. Occlusion of iliac artery did not affect the HSP72 level in muscle from sedentary rats but enhanced that in the trained rats. Mitochondrial oxidative capacity, as assessed from cytochrome oxidase and citrate synthase activities, decreased during growth in sedentary animals, but was significantly improved by endurance training. Nevertheless, increased oxidative capacity induced by endurance training was partially prevented by arterial occlusion. It is concluded that both HSP72 levels and mitochondrial oxidative capacity are affected by ischaemia and training but these changes are not necessarily related. Whereas superimposition of chronic exercise on peripheral arterial insufficiency increased HSP72 levels, our results demonstrate that endurance training even for extended period of time is not effective for improving oxidative capacity of ischaemic muscle. PMID- 11041562 TI - Assessment of mitochondrial polarization status in living cells based on analysis of the spatial heterogeneity of rhodamine 123 fluorescence staining. AB - The mitochondrial membrane potential (psimito) is an important parameter not only of mitochondrial but also of cellular status. Prolonged mitochondrial depolarization is associated with various forms of neuronal death. Assessment of mitochondrial depolarization can take advantage of the specific properties of the lipophilic dyes that distribute in a potentiometrically determined ratio across membranes. Using conventional imaging, we showed that rhodamine 123 accumulated in the mitochondria, generating a highly heterogeneous pattern of spatial distribution of fluorescence across the cell body. Collapse of the psimito following exposure to a protonophore, carbonylcyanide p chloromethoxyphenylhydrazone (CCCP), released rhodamine 123 from mitochondria into the cytosol. Under acutely changed conditions, this increased the overall intensity of the fluorescence signal and significantly decreased the degree of spatial heterogeneity of the signal. If mitochondrial depolarization was sustained chronically, the intensity of the signal decreased, but the increase in the spatial homogeneity of the fluorescent signal was maintained. Image analysis showed that the level of spatial heterogeneity of the signal can be assessed by calculating, for each individual neurone, the spread of pixel intensities values around the mean. This spread is defined by the coefficient of variation (CV), which is a measure of the standard deviation normalized to the average, and was inversely related to mitochondrial depolarization measured under different conditions. Thus, the degree of spatial heterogeneity of the rhodamine 123 signal measured from a neurone is a reliable indicator for the assessment of mitochondrial depolarization and can be used in experiments to monitor psimito over shorter or longer periods. PMID- 11041561 TI - Properties of the inward-rectifying Cl- channel in rat choroid plexus: regulation by intracellular messengers and inhibition by divalent cations. AB - The properties of the inward-rectifying Cl- conductance in rat choroid plexus epithelial cells were investigated to allow comparisons to be made with ClC-2. All experiments were performed using the whole-cell configuration of the patch clamp method. The conductance was transiently activated using an electrode solution which contained 375 nM catalytic subunit of protein kinase A (PKA). PKA failed to activate the conductance, however, when cells were pre-incubated with phorbol esters, which activate protein kinase C [1 microM phorbol 12-myristate 13 acetate (PMA) and 1 microM phorbol 12,13-dibutyrate (PDBu)]. Sustained activation of the conductance by PKA was observed in Ca2+-free conditions (5 mM BAPTA in the electrode solution), or when 100 nM calphostin C, a PKC inhibitor, was added to the electrode solution. The inward-rectifying Cl- conductance in choroid plexus is therefore similar to ClC-2 in that it is inhibited by PKC. The inward rectifying conductance was blocked when Cd2+ (30 and 300 microM) and Zn2+ (1, 30 and 300 microM) were added to the bath solution. ClC-2 channels are also blocked by Zn2+ and Cd2+. The magnitude of the inward conductance was dependent on the concentration of ATP in the electrode solution. The conductance was not observed when ATP in the electrode was replaced with non-hydrolysable ATP analogues [adenosine 5'-O-(3-thiotriphosphate) (ATP[gamma-S]) and 5' adenylylimidodiphosphate (AMP-PNP)), but it was supported by UTP and GTP. These data contrast with those of previous studies in which ClC-2 channels were activated in the absence of ATP. In conclusion, the inward-rectifying Cl- channel in rat choroid plexus shares some properties with ClC-2 (inhibition by PKC and block by divalent cations), but differs in that it depends on intracellular ATP. PMID- 11041563 TI - High-dose chemotherapy for breast cancer. PMID- 11041560 TI - The role of tumor necrosis factor (TNF) in the febrile and metabolic responses of rats to intraperitoneal injection of a high dose of lipopolysaccharide. AB - The role of tumor necrosis factor (TNF) in the febrile and metabolic responses of rats to intraperitoneal injection of a high dose of lipopolysaccharide Injection of a high dose of lipopolysaccharide (LPS) induces a septic-shock-like state, which can be accompanied by phases of hypothermia and phases of fever. In the present study we monitored body core temperature and locomotor activity, both by remote radiotelemetry, as well as changes in food intake, body mass and water intake for 3 days after an intraperitoneal (i.p.) injection of a high dose of LPS (5 mg/kg) along with sterile 0.9% saline or a neutralizing form of the soluble tumor necrosis factor (TNF) type 1 receptor (referred to as TNF-binding protein, TNF bp). Intraperitoneal injection of LPS rapidly induced high concentrations of TNF in the plasma and peritoneal lavage fluid. TNF was undetectable in the plasma and peritoneal lavage fluid of animals co-injected with LPS and TNF bp, implying neutralization of peripheral bioactive TNF. Administration of LPS induced hypothermia by about 1.5 degrees C, which lasted for 5 h after injection. During the light-time periods of days 2 and 3 after injection, the rats developed a robust fever. Treatment with TNF bp resulted in a faster recovery from the LPS induced hypothermia so that the rats developed a pronounced fever on the day of injection. Locomotor activity during night-time periods was suppressed in LPS treated animals. The LPS-induced depression of night-time activity was not antagonized by co-injection of TNF bp. On day 1 after the injection of LPS, food intake reduced to virtually zero, water intake fell to about 30% of the control value and body mass dropped by 25 g (about 10% of total body mass). With the exception of body mass, these variables recovered slowly during days 2 and 3 after LPS injection, but did not reach the control values. The LPS-induced decreases in food intake, body mass and water intake were significantly attenuated by the treatment with TNF bp. These results confirm that TNF contributes significantly to the rats' responses to intraperitoneal injection of a high dose of LPS. The fact that treatment with TNF bp accelerated and improved the rats' ability to develop a febrile response supports the view that the fever is beneficial, since all other metabolic responses measured in this study were normalized more effectively in those rats that developed a faster and more pronounced increase in body temperature. PMID- 11041564 TI - Tacrolimus clearance is age-dependent within the pediatric population. AB - For prevention of graft-versus-host disease, the consensus initial intravenous dose of tacrolimus for adults is 0.03 mg/kg/day. Whether target whole blood concentrations of tacrolimus in children undergoing hematopoietic stem cell transplantation can be achieved reproducibly with this dose is not known. We reviewed the tacrolimus blood levels and calculated clearances for 55 children (aged 6 months to 18 years, median 9 years) using tacrolimus after allogeneic marrow, blood stem cell or cord blood transplantation. The tacrolimus dose regimen was 0.03 mg/kg/day by continuous infusion starting on day -1 or day -2. At the first sampling in the peritransplant period, 71% of the tacrolimus blood levels were within the target range of 5-15 ng/ml, 87% were in the safe range of 5-20 ng/ml, 9% were toxic, and 4% were subtherapeutic. Twenty-five children were converted to oral drug using the recommended oral/intravenous dose ratio of 4.0. At the first sampling after oral conversion, 80% were in the target range, and 20% were subtherapeutic. Clearance of tacrolimus was calculated from the blood levels for patients during intravenous dosing and normalized by ideal body weight. There was a decreased clearance over the first 2 weeks only for the children >12 years old (P = 0.014). The initial calculated clearances of tacrolimus did not differ between age groups, but at steady state the mean tacrolimus clearance (+/- s.d.) was higher for those <6 years old (0.159+/-0.082 l/h/kg) than for those 6-12 years old (0.109+/-0.053 l/h/kg) or >12 years old (0.104 +/-0.068 l/h/kg). Children <6 years old undergoing hematopoietic stem cell transplantation have a higher weight-normalized tacrolimus clearance than older children and adults, and careful therapeutic monitoring is needed in the first 2 weeks after transplantation to avoid prolonged subtherapeutic dosing for this age group. PMID- 11041565 TI - Busulfan pharmacokinetics do not predict relapse in acute myeloid leukemia. AB - The purpose of this study was to evaluate the influence of busulfan (BU) pharmacokinetics on survival, grades II-IV acute graft-versus-host disease (GVHD), non-relapse mortality (NRM) and relapse in a group composed of 45 children (<18 years) and seven adults with acute myeloid leukemia (AML) in first complete remission and undergoing hematopoietic stem cell transplantation (SCT). Fifty-two patients underwent autologous (n = 25) or allogeneic (n = 27) SCT. The median age was 8.9 years (range 0.6-53 years). Conditioning therapy consisted of BU and cyclophosphamide. Improved disease-free survival was found in those patients with a steady-state concentration of BU (CssBU) below the median (<578 mg/ml, P = 0.05), and the same trend was noted for overall survival (P = 0.07). This was secondary to a higher incidence of NRM in the group of patients with CssBU above the median (P = 0.06). There was no significant correlation with CssBU and relapse (P = 0.31). No association between CssBU and GVHD was found in allogeneic patients (P = 0.30). Relapse was evaluated among the subgroups of age (< or >10 years) and transplant type (allogeneic or autologous) with no statistically significant association observed among these factors. Multiple regression analysis for relapse revealed no significant correlation with CssBU above or below the median, age, or transplant type. In this study, CssBU below the median did not correlate with an inferior outcome for patients with AML. Pharmacokinetic dosing of BU may be important for prevention of NRM but does not appear to influence the risk of relapse in this largely pediatric population with AML. PMID- 11041566 TI - Allogeneic stem cell transplantation with fludarabine-based, less intensive conditioning regimens as adoptive immunotherapy in advanced Hodgkin's disease. AB - Six patients with advanced Hodgkin's disease in which multiple conventional treatments (median prior chemotherapy regimens: seven), radiation therapy, and a prior autologous stem cell transplantation (SCT) had failed underwent allogeneic SCT following a fludarabine-based conditioning regimen. Median age was 29 years (22-30). Median time to progression after autologous SCT was 6 months (4-21). Disease status at transplant was refractory relapse (n = 3) and sensitive relapse (n = 3). Cell source was filgrastim-mobilized peripheral blood stem cells from an HLA-identical sibling (n = 4) or matched unrelated donor marrow (n = 2). Conditioning regimens were fludarabine-cyclophosphamide-antithymocyte globulin (n = 4), fludarabine-melphalan (n = 1) and fludarabine-cytarabine-idarubicin (n = 1). Myeloid recovery was prompt, with an absolute neutrophil count > or =500/microl on day 12 (11-15). Median platelet recovery to > or =20000/microl was on day 9 (0-60). Chimerism studies on day 30 indicated 100% donor-derived hematopoiesis in 4/5 evaluable patients (4/4 non-progressors). All responders (3/3) have ongoing 100% donor-derived chimerism. Acute graft-versus-host disease (GVHD) was diagnosed in 4/6 evaluable patients. Chronic GVHD was present in 2/4 evaluable patients. There were no regimen-related deaths. Overall day 100 transplant-related mortality was 2/6 (33%). Three patients have expired and three are alive and progression-free with a median follow-up of 9 months (6-26) post transplant. We conclude that allogeneic stem cell transplantation with fludarabine-based preparative regimens is feasible in these high-risk, heavily pretreated HD patients. PMID- 11041567 TI - Prolonged survival after intensive therapy and purged ABMT in patients with multiple myeloma. AB - Despite numerous strategies, the cure of multiple myeloma remains a difficult challenge. Recent approaches have involved dose-intensive therapy followed by stem cell transplantation, most often with autologous stem cells (ASCT). Although ASCT is of benefit, it is not considered curative. Between 1988 and 1995, we utilized an aggressive three-drug conditioning regimen followed by ABMT using marrow purged with either 4-hydroperoxycyclophosphamide (4-HC) or mafosphamide (MAF). Twenty-nine of 42 patients who had first received VAD (14 patients) or VAD followed by cyclophosphamide (7 g/m2 i.v.) + dexamethasone (40 mg/day p.o. x4) + GM-CSF (15 patients) met the eligibility criteria needed to undergo bone marrow harvest and ABMT, ie < or =10% marrow plasma cells and > or =50% decrease in paraprotein level. Alpha-interferon maintenance therapy was given post ABMT. Median follow-up is 7.5 years (range 5.0-11.25). Six early and two late non relapse deaths occurred; 15 patients have relapsed. Seven patients remain in continuous CR (five) or PR (two), including three with stage IIIB disease at diagnosis. One patient developed a soft tissue sarcoma 8 years post ASCT. Although this protocol produced excessive toxicity compared with current approaches, the results demonstrate that dose-intensive therapy and ASCT can produce durable remission in this disease. Further development of dose-intensive strategies is warranted. PMID- 11041568 TI - Veno-occlusive disease of the liver after allogeneic bone marrow transplantation for severe aplastic anemia. AB - There are few reports about the occurrence of hepatic VOD after BMT for severe aplastic anemia (SAA). We prospectively studied 17 patients with SAA after allogeneic BMT for the occurrence and severity of VOD. Plasma levels of protein C, protein S, antithrombin III, vWF, t-PA and PAI-1 were determined before preparative chemotherapy, on the day of marrow infusion, and on days 7, 14 and 21. VOD occurred in seven patients (41.2%) at a median of day 1 (range, day -2 to 15). Five had mild, and two moderate VOD. Platelet transfusion requirements were higher in the patients with VOD. The plasma levels of natural anticoagulants such as protein C, free protein S and antithrombin III decreased significantly on day 0 from the baseline levels. Plasma levels of t-PA, PAI-1 and vWF increased significantly in the early post-transplant period compared to the baseline levels. The mean plasma levels of t-PA on day 7 (P = 0.016) and PAI-1 on days 0 and 7 (P = 0.016, 0.032) were higher in the patients with VOD. In summary, we observed hypercoagulability and a high incidence of VOD after allogeneic BMT for SAA. Levels of t-PA and PAI-1 were significantly higher in the patients with VOD after BMT. PMID- 11041569 TI - Economic analysis of a phase III study of G-CSF vs placebo following allogeneic blood stem cell transplantation. AB - Hematopoietic colony-stimulating factors (CSF) decrease the duration of neutropenia following stem cell transplantation (SCT). With CSF-mobilized allogeneic blood SCT (alloBSCT), the yields of CD34+ cells are several-fold higher than in other SCT settings, raising concern that post-transplant CSF use may be unnecessary. In this study, we estimate the resource and cost implications associated with CSF use following alloBSCT. A cost identification analysis was conducted for 44 patients on a randomized, double-blind placebo-controlled trial of G-CSF following alloBSCT. Study drug was given daily until an absolute neutrophil count (ANC) > or = 1000 cells/microl. Billing information from the time of transplant to day +100 was analyzed. The median number of days to an ANC > or = 500 cells/microl was shorter in the G-CSF arm, 10.5 days vs 15 days (P < 0.001), while platelet recovery and rates of acute graft-versus-host disease (GVHD) and survival were similar. Resource use was similar, including days hospitalized, days on antibiotics, blood products transfused and outpatient visits. Total median post-transplant costs were $76577 for G-CSF patients and $78799 for placebo patients (P = 0.93). G-CSF following allogeneic blood SCT decreased the median duration of absolute neutropenia and did not incur additional costs, but did not result in shorter hospitalizations, or less frequent antibiotic use. PMID- 11041570 TI - Juvenile myelomonocytic leukemia relapsing after allogeneic bone marrow transplantation successfully treated with interferon-alpha. AB - We report a 5-year-old boy with juvenile myelomonocytic leukemia (JMML) which relapsed after an allogeneic bone marrow transplant who was successfully treated with interferon-alpha (IFN-alpha). One year after starting the therapy, he remains clinically well and in complete remission while continuing treatment with IFN-alpha and bestatin. Although the precise mechanism by which remission was induced is uncertain, a GVL effect combined with a direct antileukemia effect of IFN-alpha may be responsible. Further assessment of the role of IFN-alpha in relapsed JMLL patients is warranted. PMID- 11041571 TI - Myopericarditis caused by cyclophosphamide used to mobilize peripheral blood stem cells in a myeloma patient with renal failure. AB - Cyclophosphamide (CPA) is widely used for peripheral blood stem cell mobilization, and a dose adjustment of CPA in the presence of renal failure has not been suggested. However, we describe a myeloma patient with renal failure (serum creatinine 4.2 mg/dl, creatinine clearance 11.2 ml/min) receiving CPA 2 g/m2 for 2 days, who developed unexpectedly severe toxicity, including myopericarditis and prolonged myelosuppression. The serial serum concentrations of CPA metabolites were persistently much higher than those in a myeloma patient with normal renal function. We consider, therefore, that the dose of CPA should be reduced in the presence of severe renal failure when used as high-dose therapy or to mobilize peripheral blood stem cells. PMID- 11041572 TI - Successful bone marrow transplantation in a child with red blood cell pyruvate kinase deficiency. AB - We report the first successful use of BMT for the treatment of RBC pyruvate kinase (PK) deficiency in a boy who developed neonatal jaundice and severe transfusion-dependent hemolytic anemia a few months after birth. He received a BMT at the age of 5 from an HLA-identical sister who has normal PK activity after conditioning with busulfan and cyclophosphamide. The post-transplant course was uneventful. At present, 3 years after transplant, he is 8 years old and has a normal hemoglobin level and normal RBC PK activity without evidence of hemolysis. DNA analysis has confirmed full engraftment. PMID- 11041573 TI - Rapid onset retinopathy in a diabetic patient following bone marrow transplantation. AB - We report a 38-year-old man who presented in 1998 with advanced multiple myeloma and newly diagnosed diabetes mellitus (DM). Subsequent BMT has been successful after conditioning with melphalan and total body irradiation, but significant ischaemic retinopathy has developed. Chemotherapeutic agents, total body irradiation, and DM are likely to have been co-factors in precipitating the rapid onset of retinopathy. Routine ophthalmic surveillance is recommended for all patients after BMT, particularly for those with additional risk factors for the development of retinopathy such as DM. PMID- 11041574 TI - Fatal invasive cerebral Absidia corymbifera infection following bone marrow transplantation. AB - A 56-year-old dairy farmer received a fully HLA matched unrelated donor marrow transplant for high risk CML in chronic phase. His early post-transplant course was complicated by a series of massive intracerebral bleeds and by sepsis related to a malignant otitis externa. The microbial pathogen isolated from ear swabs was found to be Absidia corymbifera, but CT scan at the time showed no intracerebral extension. Despite neutrophil engraftment and aggressive antifungal management he succumbed. Autopsy revealed invasion of Absidia into the brain from the ear. We speculate that colonisation by Absidia resulted from occupational exposure. PMID- 11041575 TI - Sciatic nerve compression following bone marrow harvest. AB - We describe a donor who suffered pain secondary to sacral plexus and sciatic nerve compression post bone marrow harvest. Haematoma was demonstrated by magnetic resonance image (MRI) scanning. To our knowledge, this is the first reported case of compression neuropathy post bone marrow harvest documented by MRI scanning. Given the increasing number of bone marrow transplants being performed and the paramount importance of donor safety, compressive neuropathies need to be remembered as rare but debilitating complications of bone marrow harvesting. MRI scanning is a useful modality to investigate severe or neuropathic pain post bone marrow harvest. PMID- 11041576 TI - Persistent immunological changes 55 months after PBPct. Is the restoration of immune function possible with a longer follow-up? PMID- 11041577 TI - The management of intracapsular fractures of the proximal femur. AB - The optimum choice of treatment for an intracapsular fracture cannot be based purely on the radiological appearance of the fracture and on the age of the patient. Although these are the main considerations many other factors need to be evaluated for each individual patient. Figure 1 gives a flow diagram which helps to aid decision in treatment. The intracapsular fracture should not be thought of as the unsolved fracture. Internal fixation is indicated for selected fractures. Some require arthroplasty and for others either treatment can be used. The clinician must assess each of the individual risk factors for healing in each patient, and then decide if the risk of failure of internal fixation is high enough to justify replacing the femoral head with an arthroplasty. PMID- 11041578 TI - Thoughts on the impact of technology on orthopaedics. PMID- 11041579 TI - The Trent regional arthroplasty study. Experiences with a hip register. AB - We have assessed the usefulness of a regional hip register in the assessment of the outcome of primary total hip replacement (THR). Over 97% of THRs performed in the Trent region in 1990 were captured onto the register and the inaccuracies recorded were less than 1.8%. In an independent assessment of 2111 patients five years after THR, 85.9% of those available for assessment responded, and 66.8% agreed to an assessment. The cost of this independent assessment at five years, utilising a regional hip register, was approximately l50 per implant. This is a reasonable outlay to identify problems early. Some form of registration and outcome assessment should be performed on a national level. PMID- 11041580 TI - Comparison of in vivo wear between polyethylene liners articulating with ceramic and cobalt-chrome femoral heads. AB - At yearly intervals we compared the radiological wear characteristics of 81 alumina ceramic femoral heads with a well-matched group of 43 cobalt-chrome femoral heads. Using a computer-assisted measurement system we assessed two dimensional penetration of the head into the polyethylene liner. We used linear regression analysis of temporal data of the penetration of the head to calculate the true rates of polyethylene wear for both groups. At a mean of seven years the true rate of wear of the ceramic group was slightly greater (0.09 mm/year, SD 0.07) than that of the cobalt-chrome group (0.07 mm/year, SD 0.04). Despite the numerous theoretical advantages of ceramic over cobalt-chrome femoral heads, the wear performance in vivo of these components was similar. PMID- 11041581 TI - A conservative femoral replacement for total hip arthroplasty. A prospective study. AB - Between 1985 and 1993, 146 patients (162 hips) had total hip replacement (THR) using a conservative uncemented femoral component. The mean age of the patients was 50.8 years and the mean follow-up was 6.2 years (2 to 13). One patient was lost to follow-up, one died within two years of surgery and one had a revision procedure after a fracture sustained in a road-traffic accident. For the remaining 159, Kaplan-Meier survival analysis was calculated for the incidence of revision because of mechanical loosening or osteolysis. Survival without mechanical loosening at both five and ten years was 98.2%. Survival without osteolysis was 99% at five and 91% at ten years. The Harris hip score improved from a mean of 66.3 before to 90.4 at follow-up. Of particular note is the lack of thigh pain in this group. Radiological analysis showed that 139 stems (88%) had no measurable subsidence, 8 (5%) had less than 2 mm and 12 (7%) had more than 2 mm. Two of the eight and one of the 12 were revised for mechanical loosening. Nine hips were revised for late loosening associated with osteolysis. No reaming of the femoral canal was associated with statistically significant less blood loss compared with a comparable control group of uncemented implants (p < 0.0001). Our study suggests that using a conservative femoral implant does not protect against wear debris but the reliable mechanical stability (98.2%) makes this an attractive design of implant particularly for young patients. PMID- 11041582 TI - Fix and flap: the radical orthopaedic and plastic treatment of severe open fractures of the tibia. AB - We performed a retrospective review of the case notes of 84 consecutive patients who had suffered a severe (Gustilo IIIb or IIIc) open fracture of the tibia after blunt trauma between 1990 and 1998. All had been treated by a radical protocol which included early soft-tissue cover with a muscle flap by a combined orthopaedic and plastic surgery service. Our ideal management is a radical debridement of the wound outside the zone of injury, skeletal stabilisation and early soft-tissue cover with a vascularised muscle flap. All patients were followed clinically and radiologically to union or for one year. After exclusion of four patients (one unrelated death and three patients lost to follow-up), we reviewed 80 patients with 84 fractures. There were 67 men and 13 women with a mean age of 37 years (3 to 89). Five injuries were grade IIIc and 79 grade IIIb; 12 were site 41, 43 were site 42 and 29 were site 43. Debridement and stabilisation of the fracture were invariably performed immediately. In 33 cases the soft-tissue reconstruction was also completed in a single stage, while in a further 30 it was achieved within 72 hours. In the remaining 21 there was a delay beyond 72 hours, often for critical reasons unrelated to the limb injury. All grade-IIIc injuries underwent immediate vascular reconstruction, with an immediate cover by a flap in two. All were salvaged. There were four amputations, one early, one mid-term and two late, giving a final rate of limb salvage of 95%. Overall, nine pedicled and 75 free muscle flaps were used; the rate of flap failure was 3.5%. Stabilisation of the fracture was achieved with 19 external and 65 internal fixation devices (nails or plates). Three patients had significant segmental defects and required bone-transport procedures to achieve bony union. Of the rest, 51 fractures (66%) progressed to primary bony union while 26 (34%) required a bone-stimulating procedure to achieve this outcome. Overall, there was a rate of superficial infection of the skin graft of 6%, of deep infection at the site of the fracture of 9.5%, and of serious pin-track infection of 37% in the external fixator group. At final review all patients were walking freely on united fractures with no evidence of infection. The treatment of these very severe injuries by an aggressive combined orthopaedic and plastic surgical approach provides good results; immediate internal fixation and healthy soft tissue cover with a muscle flap is safe. Indeed, delay in cover (>72 hours) was associated with most of the problems. External fixation was associated with practical difficulties for the plastic surgeons, a number of chronic pin-track infections and our only cases of malunion. We prefer to use internal fixation. We recommend primary referral to a specialist centre whenever possible. If local factors prevent this we suggest that after discussion with the relevant centre, initial debridement and bridging external fixation, followed by transfer, is the safest procedure. PMID- 11041583 TI - Late fracture of the hip after reamed intramedullary nailing of the femur. AB - In a consecutive series of 498 patients with 528 fractures of the femur treated by conventional interlocking intramedullary nailing, 14 fractures of the femoral neck (2.7%) occurred in 13 patients. The fracture of the hip was not apparent either before operation or on the immediate postoperative radiographs. It was diagnosed in the first two weeks after operation in three patients and after three months in the remainder. Age over 60 years at the time of the femoral fracture and female gender were significantly predictive of hip fracture on bivariate logistic regression analysis, but on multivariate analysis only the location of the original fracture in the proximal third of the femur (p = 0.0022, odds ratio = 6.96, 95% CI 2.01 to 24.14), low-energy transfer (p = 0.0264, odds ratio = 15.56, 95% CI 1.38 to 75.48) and the severity of osteopenia on radiographs (p = 0.0128, odds ratio = 7.55, 95% CI 1.54 to 37.07) were significant independent predictors of later fracture. Five of the 19 women aged over 60 years, who sustained an osteoporotic proximal diaphyseal fracture of the femur during a simple fall, subsequently developed a fracture of the neck. Eleven of the hip fractures were displaced and intracapsular and, in view of the advanced age of most of these patients, were usually treated by replacement arthroplasty. Reduction and internal fixation was used to treat the remaining three intertrochanteric fractures. Three patients developed complications requiring further surgery; five died within two years of their fracture. PMID- 11041584 TI - The role of physiotherapy and clinical predictors of outcome after fracture of the distal radius. AB - The capacity for physiotherapy to improve the outcome after fracture of the distal radius is unproven. We carried out a randomised controlled trial on 96 patients, comparing conventional physiotherapy with a regime of home exercises. The function of the upper limb was assessed at the time of removal of the plaster cast and at three and six months after injury. Factors which may predict poor outcome in these patients were sought. Grip strength and hand function did not significantly differ between the two groups. Flexion and extension of the wrist were the only movements to improve with physiotherapy at six months (p = 0.001). Predictors of poor functional outcome were malunion and impaired function before the fracture. These patients presented with pain, decreased rotation of the forearm and low functional scores at six weeks. Our study has shown that home exercises are adequate rehabilitation after uncomplicated fracture of the distal radius, and routine referral for a course of physiotherapy should be discouraged. The role of physiotherapy in patients at high risk of a poor outcome requires further investigation. PMID- 11041585 TI - Nonunion of the humerus after failure of surgical treatment. Management using the Ilizarov circular fixator. AB - We used the Ilizarov circular external fixator to treat 16 patients with persistent nonunion of the diaphysis of the humerus despite surgical treatment. All patients had pain and severe functional impairment of the affected arm. In ten, nonunion followed intramedullary nailing. We successfully treated these by a closed technique. The nail was left in place and the fracture compressed over it. The fractures of the other six patients had previously been fixed by various methods. We explored these nonunions, removed the fixation devices and excised fibrous tissue and dead bone before stabilising with the Ilizarov fixator. In five patients union was achieved. Bone grafting was not required. In the single patient in whom treatment failed, there had been a severely comminuted open fracture. All except one patient had reduction of pain, and all reported an improvement in function. PMID- 11041586 TI - Endoscopic reconstruction of the anterior cruciate ligament with an ipsilateral patellar tendon autograft. A prospective longitudinal five-year study. AB - A total of 90 patients with an isolated rupture of the anterior cruciate ligament (ACL) had a reconstruction using the ipsilateral patellar tendon secured with round-headed cannulated interference screws. Annual review for five years showed three failures of the graft (two traumatic and one atraumatic); none occurred after two years. Ten patients sustained a rupture of the contralateral ACL. At five years, 69% of those with surviving grafts continued to participate in moderate to strenuous activity. Using the International Knee Documentation Committee assessment, 90% reported their knee as being normal or nearly normal and had a median Lysholm knee score of 96 (64 to 100). Most patients (98%) had a pivot shift of grade 0 with the remaining 2% being grade 1; 90% of the group had a Lachman test of grade 0. The incidence of subsequent meniscectomy was similar in the reconstructed joint to that in the contralateral knee. Radiological examination was normal in 63 of 65 patients. Our study supports the view that reconstruction of the ACL is a reliable technique allowing full rehabilitation of the previously injured knee. In the presence of normal menisci there is a low incidence of osteoarthritic change despite continued participation in sporting activity. PMID- 11041587 TI - A modified Thompson quadricepsplasty for the stiff knee. AB - Between March 1987 and March 1997, we performed a modified Thompson quadricepsplasty on 20 stiff knees and followed the patients for a mean of 35 months (24 to 52). After the operation, the knee was immobilised in flexion and periodically extended. At the final follow-up, the mean active flexion was 113.5 degrees (75 to 150). The final mean gain in movement was 67.6 degrees (5 to 105). One patient had a deep infection which resolved after wound care and intravenous antibiotics. The modified Thompson quadricepsplasty with appropriate postoperative care can give good results. PMID- 11041588 TI - Radiological changes five years after unicompartmental knee replacement. AB - Failure of a unicompartmental knee replacement (UKR) may be caused by progressive osteoarthritis of the knee and/or failure of the prosthesis. Limb alignment can influence both of these factors. We have examined the fate of the other compartments and measured changes in leg alignment after UKR. A total of 50 UKRs was carried out on 45 carefully selected patients between 1989 and 1992. At operation, deliberate attempts were made to avoid overcorrection of the deformity. Four patients died, one patient was lost to follow-up and two knees were revised before review which was at a minimum of five years. Standard long leg weight-bearing anteroposterior views of the knee and skyline views of the patellofemoral joint were taken before and at eight months and five years after operation. The radiographs of the remaining 43 knees were reviewed twice by blind and randomised assessment to measure the progression of osteoarthritis within the joints. Overcorrection of the deformity in the coronal plane was avoided in all but two knees. Only one showed evidence of progression of osteoarthritis within the patellofemoral joint, and this was only identified in one of the four assessments. Deterioration in the state of the opposite tibiofemoral compartment was not seen. Varus deformity tended to recur. Recurrent varus of 2 degrees was observed between eight months and five years after operation. There was no correlation between the postoperative tibiofemoral angle and the extent of recurrent varus recorded at five years. Changes in alignment may be indicative of minor polyethylene wear or of subsidence of the tibial component. The incidence of progressive osteoarthritis within the knee was very low after UKR. Patients should be carefully selected and overcorrection of the deformity be avoided. PMID- 11041589 TI - Improvement in function after valgus bracing of the knee. An analysis of gait symmetry. AB - The use of a valgus brace can effectively relieve the symptoms of unicompartmental osteoarthritis of the knee. This study provides an objective measurement of function by analysis of gait symmetry. This was measured in 30 patients on four separate occasions: immediately before and after initial fitting and then again at three months with the brace on and off. All patients reported immediate symptomatic improvement with less pain on walking. After fitting the brace, symmetry indices of stance and the swing phase of gait showed a consistent and immediate improvement at 0 and 3 months, respectively, of 3.92% (p = 0.030) and 3.40% (p = 0.025) in the stance phase and 11.78% (p = 0.020) and 9.58% (p = 0.005) in the swing phase. This was confirmed by a significant improvement at three months in the mean Hospital for Special Surgery (HSS) knee score from 69.9 to 82.0 (p < 0.001). Thus, wearing a valgus brace gives a significant and immediate improvement in the function of patients with unicompartmental osteoarthritis of the knee, as measured by analysis of gait symmetry. PMID- 11041590 TI - Osteoarticular allograft in surgery for high-grade malignant tumours of bone. AB - We assessed the results of 17 limb-salvage procedures using osteoarticular allografts after wide resection of high-grade malignant bone tumours. All patients received chemotherapy. At the five-year follow-up, three patients had died from metastases. The allografts survived for five years in only seven patients all of whom had good function, ranging from 73% to 90% of normal. The allografts were removed because of fracture in seven patients and infection in one, and in all of these a second limb-salvage procedure was undertaken. With such a low rate of survival of osteoarticular allografts, we believe that their use in the management of high-grade malignant bone tumours should, at best, be considered a temporary solution. PMID- 11041591 TI - Wrist arthroscopy without distraction. A technique to visualise instability of the wrist after a ligamentous tear. AB - We describe a technique for arthroscopy of the wrist which is carried out without traction and with the arm lying horizontally on the operating table. The wrist is not immobilised, which makes it possible to assess the extent of instability after a ligamentous tear. In a prospective study of 30 patients we compared this technique with conventional wrist arthroscopy, performing the new method first followed by conventional arthroscopy. The advantages are that the horizontal position of the arm allows the surgeon to proceed directly from arthroscopic diagnosis to treatment, and that no change of position is required for fluoroscopy. In terms of diagnostic sensitivity, we found our technique matched that of conventional arthroscopy. We had no difficulty in carrying out minor surgical procedures such as debridement and suturing. PMID- 11041592 TI - The Sauve-Kapandji procedure for post-traumatic disorders of the distal radio ulnar joint. AB - We present the results of a retrospective series of 41 Sauve-Kapandji procedures carried out for complications of fractures of the distal radius. All the operations were undertaken by one surgeon with a mean follow-up of 32 months. A total of 37 patients was available for clinical review. The indications for surgery were pain on the ulnar side of the wrist and decreased rotation of the forearm. Intraperiosteal and extraperiosteal techniques were used for resection of the ulna, with no difference in outcome. Patients were assessed for pain, rotation of the forearm and complications. A Mayo Modified Wrist Score was used. Pain was improved in 25 of the 37 patients, and unchanged in ten. Rotation of the forearm returned to within 7 degrees of the uninjured side. The results are discussed in relation to the presence of preoperative malunion of the distal radius, age and the functional outcome. Age is not a contraindication for this procedure. PMID- 11041593 TI - Arthroscopic treatment for impingement of the anterolateral soft tissues of the ankle. AB - We treated 52 patients with impingement of the anterolateral soft tissues of the ankle by arthroscopic debridement. All had a history of single or multiple inversion injuries, without instability. One half had negative stress radiographs (stable group), while the others were positive (unstable group). Their mean age was 31 years and there were 35 men and 17 women. The results were assessed at a mean follow-up of 30 months. Three patients (6%) had a fair result, while 49 (94%) had an excellent or good outcome. No difference was found in the final results between the two groups (p > 0.05). We conclude that anterolateral impingement of the ankle should be considered in a patient with chronic anterolateral pain after an injury, regardless of the stability of the ankle. PMID- 11041594 TI - Vascularised fibular grafts. An experience of 102 patients. AB - The results and complications of 104 vascularised fibular grafts in 102 patients are presented. Bony union was ultimately achieved in 97 patients, with primary union in 84 (84%). The mean time to union was 15.5 weeks (8 to 40). In 13 patients, primary union was achieved at one end of the fibula and secondary union at the other end. In these patients, the mean time to union was 31.1 weeks (24 to 40). Five patients failed to achieve union, with a resultant pseudarthrosis (3 patients) or amputation (2 patients). There were various complications. Immediate thrombosis occurred in 14 cases. In two of 23 patients with osteomyelitis, infection recurred at two and six months after surgery, respectively. Both patients had active osteomyelitis less than one month before the operation. Bony infection occurred in a patient with a synovial sarcoma of the forearm one year after surgery. In 15 patients, 19 fractures of the fibular graft occurred after bony union, all except one within one year after union. In patients in whom an external fixator had been used, fracture occurred soon after its removal. Union was difficult to achieve in cases of congenital pseudarthrosis of the tibia. Appropriate alignment of the fibular graft is an important factor in preventing stress fracture. The vascularised fibula should be protected during the first year after union. Postoperative complications at the donor site included transient palsy of the superficial peroneal nerve in three patients, contracture of flexor hallucis longus in two and valgus deformity of the ankle in three. Vascularised fibular grafts are useful in the reconstruction of massive bony defects. We believe that meticulous preoperative planning, including choosing which vessels to select in the recipient and the type of fixation devices to use, and care in the introduction of the vascularised fibula, can improve the results and prevent complications. PMID- 11041595 TI - Serrated W/M osteotomy. Results using a new technique for the correction of infantile tibia vara. AB - The conventional osteotomies used to treat infantile tibia vara (Blount's disease) may require internal fixation and its subsequent removal. These techniques, which carry the risk of traction injury, and potential problems of stability and consolidation, do not always succeed in correcting the rotational deformity which accompanies the angular deformity. We have used a new surgical approach, the serrated W/M osteotomy of the proximal tibia, to correct infantile tibia vara in 15 knees of 11 patients. We present the results in 13 knees of nine patients who have been followed up for a mean of eight years. The mean angular correction achieved after operation was 18 +/- 5.8 degrees. The mean femorotibial shaft angle was corrected from 14.2 +/- 3.7 degrees of varus to 4.6 +/- 4.4 degrees of valgus. At the last follow-up, the mean angular correction had reduced to 1.3 +/- 4.9 degrees of valgus without compromising the rotational correction and the overall good clinical results. All the patients and parents were satisfied, rating the result as excellent or good. There were no major postoperative complications and no reoperations. Eight patients were free from pain and able to perform physical activities suitable for their age. One complained of occasional pain. This procedure has the advantage of allowing both angular and rotational correction with a high degree of success without the need for internal fixation. PMID- 11041596 TI - Treatment of cubitus varus using the Ilizarov technique of distraction osteogenesis. AB - Seven children with a post-traumatic cubitus varus deformity were treated using the Ilizarov technique of distraction osteogenesis. The outcome was rated as excellent in each case and all were satisfied with the cosmetic appearance. No complications had been encountered by the latest follow-up at a mean of 66.7 months. This technique seems reliable for the treatment of such deformities, provided that it achieves full correction by gradual distraction. Nerve palsy and unsightly scars are avoided, and the range of movement of adjacent joints is preserved. PMID- 11041597 TI - Injury to the spinal cord without radiological abnormality (SCIWORA) in adults. AB - Injury to the spinal cord without radiological abnormality often occurs in the skeletally immature cervical and thoracic spine. We describe four adult patients with this diagnosis involving the cervical spine with resultant quadriparesis. The relevant literature is reviewed. The implications for initial management of the injury, the role of MRI and the need for a high index of suspicion are highlighted. PMID- 11041598 TI - Instability of the coccyx in coccydynia. AB - Coccygectomy is a controversial operation. Some authors have reported good results, but others advise against the procedure. The criteria for selection are ill-defined. We describe a study to validate an objective criterion for patient selection, namely radiological instability of the coccyx as judged by intermittent subluxation or hypermobility seen on lateral dynamic radiographs when sitting. We enrolled prospectively 37 patients with chronic pain because of coccygeal instability unrelieved by conservative treatment who were not involved in litigation. The operation was performed by the same surgeon. Patients were followed up for a minimum of two years after coccygectomy, with independent assessment at two years. There were 23 excellent, 11 good and three poor results. The mean time to definitive improvement was four to eight months. Coccygectomy gave good results in this group of patients. PMID- 11041599 TI - Subacute subdural haematoma complicating lumbar microdiscectomy. AB - There have been no previous reports of a spinal subdural haematoma occurring as a complication of spinal surgery. We highlight the pitfalls in the diagnosis and management of a subacute subdural haematoma resulting from a dural tear which occurred as a surgical complication of microdiscectomy. PMID- 11041600 TI - Hangman's fracture: the relationship between asymmetry and instability. AB - There is ambiguity concerning the nomenclature and classification of fractures of the ring of the second cervical vertebra (C2). Disruption of the pars interarticularis which defines true traumatic spondylolisthesis of C2, is often wrongly called a pedicle fracture. Our aim in this study was to assess the influence of asymmetry on the anatomical and functional outcome and to evaluate the criteria of instability established by Roy-Camille et al. We studied the plain radiographs and CT scans of 24 patients: 13 were judged to be asymmetrical, ten were considered unstable and 14 stable. Treatment was with a Minerva jacket in 15 fractures and by operation in nine. Surgery was undertaken in patients with severe C2 to C3 sprains. One patient with an unstable lesion refused operation and was treated conservatively with a poor radiological result. Our study showed that asymmetry of the fracture did not affect the outcomes of treatment and should not therefore influence decisions in treatment. The criteria of Roy Camille seem to be reliable and useful. We prefer the posterior approach to the cervical spine, which allows both stabilisation of the fracture and correction of a local kyphosis. PMID- 11041602 TI - The influence of avascularity on the mechanical properties of human bone-patellar tendon-bone grafts. AB - Our aim was to analyse the effect of avascularity on the morphology and mechanical properties (tensile strength, viscoelasticity) of human bone-patellar tendon-bone (BPTB) grafts in vitro. These were harvested at postmortem and stored submerged in denaturated human plasma at a constant pH, pO2, pCO2, temperature and humidity under sterile conditions. Mechanical testing was performed two and four weeks after removal of the graft. The mean ultimate strength was 1085.7 +/- 255.8 N (control), 1009.0 +/- 314.9 N (two weeks cultured) and 1076.8 +/- 414.8 N (four weeks cultured). There was no significant difference in linear stiffness or deformation to failure between the groups. There was a difference in viscoelasticity between the control group and the avascular grafts and the latter had significant lower peak load-to-load ratios after 15 minutes compared with the control group. After two and four weeks the graft contained viable fibroblasts. There was regular cellularity in the superficial layers and decreased cellularity in the midportion. The structure of the collagen including the crimp pattern appeared to be normal in polarised light. We conclude that avascularity does not significantly affect ultimate failure loads or stiffness of BPTB grafts. Slight changes in viscoelasticity were induced, but the significance of the increased stress relaxation is not fully understood. PMID- 11041601 TI - Modulation of the formation of adhesions during the healing of injured tendons. AB - The formation of restrictive adhesions around the musculotendinous unit after injury is one of the most vexing processes faced by the surgeon. In flexor tendons it has been shown that the synovial tissue is the source of aggressive fibroblasts which contribute to this process. Using a rabbit model, we have examined the effects of treating the synovial sheath with the antimetabolite 5 fluorouracil (5-FU) for five minutes. Inflammatory, proliferative and molecular markers were compared in the response of the treated and control tendons to injury. Compared with a control group we found that the proliferative and inflammatory responses were significantly reduced (p < 0.001) in the treated tendons. Not only was there a reduction in the cellular and cytokine response, but there also was a significant (p < 0.001) reduction in the level of activity of the known pro-scarring agent, transforming growth factor beta 1 (TGF-beta1). These pilot studies indicate that the formation of restrictive adhesions may be modulated using a simple single-touch technique in the hope of producing a better return of function. PMID- 11041603 TI - Precision of the measurements of periprosthetic bone mineral density in hips with a custom-made femoral stem. AB - Our aim was to determine the precision of the measurements of bone mineral density (BMD) by dual-energy x-ray absorptiometry in the proximal femur before and after implantation of an uncemented implant, with particular regard to the significance of retro- and prospective studies. We examined 60 patients to determine the difference in preoperative BMD between osteoarthritic and healthy hips. The results showed a preoperative BMD of the affected hip which was lower by a mean of 4% and by a maximum of 9% compared with the opposite side. In addition, measurements were made in the operated hip before and at ten days after operation to determine the effect of the implantation of an uncemented custom made femoral stem. The mean increase in the BMD was 8% and the maximum was 24%. Previous retrospective studies have reported a marked loss of BMD on the operated side. The precision of double measurements using a special foot jig showed a modified coefficient of variation of 0.6% for the non-operated side in 15 patients and of 0.6% for the operated femur in 20 patients. The effect of rotation on the precision of the measurements after implantation of an uncemented femoral stem was determined in ten explanted femora and for the operated side in ten patients at 10 degrees rotation and in 20 patients at 30 degrees rotation. Rotation within 30 degrees influenced the precision in studies in vivo and in vitro by a mean of 3% and in single cases in up to 60%. Precise prediction of the degree of loss of BMD is thus only possible in prospective cross-sectional measurements, since the effect of the difference in preoperative BMD, as well as the apparent increase in BMD after implantation of an uncemented stem, is not known from retrospective studies. The DEXA method is a reliable procedure for determining periprosthetic BMD when positioning and rotation are strictly controlled. PMID- 11041604 TI - Early reactions after reimplantation of the tendon of supraspinatus into bone. A study in rabbits. AB - In 14 rabbits we determined the origin of the cells effecting healing of the tendon of supraspinatus inserted into a bony trough. After two weeks both the cellularity of the underlying bone and the thickness of the subacromial bursa were significantly increased in the operated compared with the control shoulders. The cellularity of the stump of the tendon, however, was significantly decreased in the operated shoulders. In this model, both the underlying bone and the subacromial bursa but not the stump of the tendon contributed to the process of repair. We conclude that the medial stump should be debrided judiciously but that cutting back to bleeding tissue is not necessary during repair of the rotator cuff. Moreover, great care should be taken to preserve the subacromial bursa since it seems to play an important role in the healing process. PMID- 11041605 TI - Low-intensity pulsed ultrasound accelerates bone maturation in distraction osteogenesis in rabbits. AB - We investigated the effects of low-intensity pulsed ultrasound on distraction osteogenesis in a rabbit model. Callotasis of the right tibia was performed in 70 male Japanese white rabbits using mini-external fixators. In the first part of the study in 64 animals using normal distraction (waiting period seven days; distraction rate 0.5 mm/12 hours; distraction period ten days), we evaluated the distraction site by radiography, measurement of the bone mineral density (BMD), mechanical testing, and histology. In the second part in six rabbits using fast distraction (waiting period 0 days; distraction rate 1.5 mm/12 hours; distraction period seven days) the site was evaluated radiologically. Half of the animals (35) had received ultrasound to their right leg (30 mW/cm2) for 20 minutes daily after ceasing distraction (ultrasound group), while rigid fixation only was maintained in the other half (control group). With normal distraction, the hard callus area, as shown by radiography, the BMD, and the findings on mechanical testing, were significantly greater in those receiving ultrasound than in the control group. Histological analysis showed no tissue damage attributable to exposure to ultrasound. With fast distraction, immature bone regeneration was observed radiologically in the control group, while bone maturation was achieved in the ultrasound group. We conclude that ultrasound can accelerate bone maturation in distraction osteogenesis in rabbits, even in states of poor callotasis. PMID- 11041606 TI - At the crossroads--neonatal detection of developmental dysplasia of the hip. PMID- 11041607 TI - Thromboprophylaxis--which treatment for which patient? PMID- 11041608 TI - The Baumann procedure for fixed contracture of the gastrosoleus in cerebral palsy. PMID- 11041609 TI - Implantation of a soft-tissue expander before operation for club foot in children. PMID- 11041610 TI - Fixation of fractures of the shaft of the humerus by dynamic compression plate or intramedullary nail. PMID- 11041611 TI - Wound infection in hip and knee arthroplasty. PMID- 11041612 TI - New York Medical College. PMID- 11041613 TI - New York University School of Medicine. PMID- 11041614 TI - University of Rochester School of Medicine and Dentistry. PMID- 11041615 TI - State University of New York (SUNY) Downstate College of Medicine. PMID- 11041616 TI - State University of New York at Stony Brook Health Sciences Center. PMID- 11041617 TI - SUNY Upstate Medical University (formerly SUNY Health Science Center at Syracuse) College of Medicine. PMID- 11041618 TI - Duke University School of Medicine. PMID- 11041619 TI - The Brody School of Medicine at East Carolina University. PMID- 11041620 TI - University of North Carolina at Chapel Hill School of Medicine. PMID- 11041621 TI - Wake Forest University School of Medicine. PMID- 11041622 TI - University of North Dakota School of Medicine and Health Sciences. Medical Student Education Council. PMID- 11041623 TI - Case Western Reserve University School of Medicine. PMID- 11041624 TI - University of Cincinnati College of Medicine. PMID- 11041625 TI - Medical College of Ohio. PMID- 11041626 TI - Northeastern Ohio Universities College of Medicine. PMID- 11041627 TI - The Ohio State University College of Medicine and Public Health. PMID- 11041628 TI - Identification of complete denture problems: a summary. AB - In this section, guidelines to the diagnosis of complete denture problems are presented in tabular form. Suggestions to the management of these problems are listed. PMID- 11041629 TI - Muscle cross-bridge chemistry and force. PMID- 11041630 TI - Muscle chemistry and force. PMID- 11041631 TI - [Radiation exposure dosage in CT studies: results of a nationwide inquiry]. PMID- 11041632 TI - Anticorresponding p15 promoter methylation and microsatellite instability in acute myeloblastic leukemia. PMID- 11041633 TI - The platelet glycoprotein Ia C807T polymorphism as risk factor for coronary catheter interventions. PMID- 11041634 TI - [On establishment of the Fundamental Law of the Protection of Personal Information]. PMID- 11041635 TI - Psychosocial problems among women cohabiting with heavy smoking men. PMID- 11041636 TI - Response from members of the Griffiths inquiry. PMID- 11041637 TI - UK case of congenital rubella can be linked to Greek cases. PMID- 11041638 TI - Compulsory retirement of experts. PMID- 11041639 TI - Public health must make best use of leadership resources. PMID- 11041641 TI - Do you admit to working within the system? Culture of not telling tales has to end. PMID- 11041640 TI - Doctors must be more aware of problems of gay, lesbian, and bisexual youth. PMID- 11041642 TI - Do you admit to working within the system? If we see errors being made how can we be expected to stay silent? PMID- 11041643 TI - Lifestyle modification may be beneficial in diabetes care. PMID- 11041645 TI - Complete bibliography of Professor Dr. med. Dr. h.c.mult. Theodor M. Fliedner. PMID- 11041644 TI - Proceedings of the International Stem Cell Workshop on Pathophysiology, Diagnostic and Therapeutic Implications of Blood Stem Cells. Ulm, Germany, July 9 12, 1997. Dedicated to Theodor M. Fliedner. PMID- 11041646 TI - Strategy from the porch. . PMID- 11041647 TI - Acute coronary interventions in the elderly. Overview. PMID- 11041648 TI - Clinical governance is the latest concept behind healthcare in the UK. PMID- 11041649 TI - Chi-squared test. PMID- 11041650 TI - Renaming protein secretion in the gram-negative bacteria. PMID- 11041651 TI - Developmental anomalies of the craniocervical junction and cervical spine. AB - Developmental anomalies of the craniocervical junction and cervical spine are relatively common in childhood and are potentially dangerous because of instability, particularly in the context of trauma. It is important that the radiologist understands and identifies these anomalies to contribute to their proper management. This article reviews the common and important anomalies of the craniovertebral junction, cervical spinal column, and related craniospinal neuraxis. PMID- 11041652 TI - Stroke recovery: how the computer reprograms itself. Neuronal plasticity: the key to stroke recovery. Kananskis, Alberta, Canada, 19-22 March 2000. PMID- 11041653 TI - Legitimizing the stepchild: organizing and expanding fellowship opportunities in endoscopic surgery. PMID- 11041654 TI - The modern role of hysteroscopy in the care of women. PMID- 11041655 TI - Incidence of mullerian defects in fertile and infertile women. PMID- 11041656 TI - Should laparoscopic hysterectomy replace vaginal hysterectomy. PMID- 11041657 TI - Hysteroscopic fluid monitoring guidelines. PMID- 11041659 TI - Development of flexible culdoscopy. PMID- 11041658 TI - The AAGL classification system for laparoscopic hysterectomy. PMID- 11041660 TI - Development of flexible culdoscopy. PMID- 11041661 TI - Selected bibliography. PMID- 11041662 TI - Proceedings of the Dr. Maurice Hilleman Symposium, August 30, 1999, in honor of his 80th birthday. PMID- 11041663 TI - Has the exclusive ulnar or median innervation of the ring finger been shown not to exist? PMID- 11041664 TI - Doctors' databank may be opened. PMID- 11041665 TI - NIH celebrates a decade of women's health research. PMID- 11041667 TI - 8th Annual Congress on Women's Health and Gender-Based Medicine: New management strategies emerging from gender-based biology. Hilton Head Island, South Carolina, USA. June 3-6, 2000. Abstracts. PMID- 11041666 TI - WebWatch--Women's health and gender-based medicine. Online consultations. PMID- 11041668 TI - Index of suspicion. Case #1. Diagnosis: Nongroup A poststreptococcal reactive arthritis. PMID- 11041669 TI - Index of suspicion. Case #2. Diagnosis: Anemia from hookworm infestation. PMID- 11041670 TI - Index of suspicion. Case #3. Diagnosis: Telogen effluvium. PMID- 11041671 TI - Transplant livers in Wilson's disease for hepatic, not neurologic, indications. PMID- 11041672 TI - European Academy for Medicine of Aging. Group of European Professors of Medical Gerontology. Zion, Switzerland. Abstracts. PMID- 11041673 TI - Re: statistical considerations in the intent-to-treat principle. PMID- 11041674 TI - Hypertension chat: tips for your patients. PMID- 11041675 TI - Effect of angiotensin-converting enzyme inhibition compared with conventional therapy on cardiovascular morbidity and mortality in hypertension: the Captopril Prevention Project (CAPP) randomized trial. The Captopril Prevention Project (CAPP) Study Group. PMID- 11041676 TI - Diabetic neuropathy: diagnosis and treatment for the pain management specialist. AB - Diabetic neuropathy is the name used by clinicians to describe a heterogeneous group of diseases that affect the autonomic and peripheral nervous systems of patients suffering from diabetes mellitus. This article provides the pain management specialist with an overview of the pathophysiology and current trends in the diagnosis and treatment of these diseases. Because of the significant morbidity and suffering associated with diabetic neuropathy, this article emphasizes practical steps to prevent, treat, and manage diabetic neuropathy to assist the pain management specialist in caring for patients suffering from this common malady. PMID- 11041677 TI - Cystic fibrosis related diabetes. PMID- 11041678 TI - [Conservative treatment of gallstones: contra]. PMID- 11041679 TI - Brain plasticity and epilepsy. Papers presented at the 5th Workshop on the Neurobiology of Epilepsy (WONOEP V). Cesky Krumlov, Czech Republic. September 8 10, 1999. PMID- 11041680 TI - Generation of superoxide from nitric oxide synthase. PMID- 11041681 TI - A key negative control experiment provides evidence that nitric oxide synthase does not catalyze superoxide formation. PMID- 11041683 TI - [Hepatic arterial infusion of chemotherapy after resection of hepatic metastases from colorectal cancer]. PMID- 11041682 TI - FIGO staging classifications and clinical practice guidelines in the management of gynecologic cancers. FIGO Committee on Gynecologic Oncology. PMID- 11041684 TI - Continuing education in endoscopy: live courses or video format? PMID- 11041685 TI - [Dr. Shaul Tchernichowski: physician, poet and lover of mankind]. PMID- 11041686 TI - [Helen Brook Taussig (1898-1986)]. PMID- 11041687 TI - Cost-effectiveness of hepatitis A vaccination in patients with chronic hepatitis C. PMID- 11041688 TI - Cost-effectiveness of hepatitis A vaccination in patients with chronic hepatitis C. PMID- 11041689 TI - Comprehensive allelotype study of hepatocellular carcinoma. PMID- 11041690 TI - Acute hepatitis after starting zinc therapy in a patient with presymptomatic Wilson's disease. PMID- 11041691 TI - Difficulties in conducting controlled trials in radical therapies for nonadvanced hepatocellular carcinoma. PMID- 11041692 TI - Normalizing the prothrombin time. PMID- 11041693 TI - Boeck's sarcoidosis--a centennial. PMID- 11041695 TI - [Holistic medicine and patient care management by family practice/ specialists]. PMID- 11041694 TI - Combination of clobetasol and tretinoin in vitiligo -letter-. PMID- 11041696 TI - [The medical profession and social accountability. Opening address at the 103. German Doctors' Day in Cologne, May 9, 2000]. PMID- 11041697 TI - [What type of physician is needed by the nation? The doctor and patient in a struggle between physician's responsibility and patient's autonomy, 4th Baden Wurttemberg Doctor's Day on July 14, 2000 at Constance]. PMID- 11041698 TI - [Economization of caring. Rate setting as health care policy]. PMID- 11041699 TI - [New development in diabetes therapy. 35th Annual Meeting of the German Diabetes Society, Munich, May 5-June 3, 2000]. PMID- 11041700 TI - Prof. Sumio Umezawa, 1909-2000. Obituary. PMID- 11041701 TI - Effect of vecuronium on the release of acetylcholine after nerve stimulation. PMID- 11041702 TI - Plating techniques for the dorsum of the distal radius. PMID- 11041703 TI - Elbow revision anthroplasty. PMID- 11041704 TI - Clinical diagnostic tests for carpal tunnel syndrome. PMID- 11041705 TI - Second episode of tuberculosis in an HIV-infected child: relapse or reinfection? AB - We report a case of an HIV-infected child with a second episode of tuberculosis 22 months after completing antituberculosis treatment. DNA fingerprinting of organisms from both episodes showed an identical strain of Mycobacterium tuberculosis. We believe this to be the first case of confirmed relapsed tuberculosis in an HIV-infected child, and suggest that a longer course of antituberculosis treatment be given to such children. ? 2000 The British Infection Society. PMID- 11041706 TI - A case of costochondral abscess due to Corynebacterium minutissimum in an HIV infected patient. AB - Corynebacterium minutissimum, known as the causative agent of erythrasma, has recently been reported as a clinically significant pathogen in the immunocompromised host. We report for the first time the possible involvement of a multidrug-resistant C. minutissimum strain in a costochondral abscess occurring in an HIV-infected patient. PMID- 11041707 TI - Orbital hydatid cyst: treatment and prevention of recurrences with albendazole plus praziquantel. AB - A case of successful treatment of orbital echinococcosis without evidence of recurrence on prolonged follow-up is presented. The management of orbital hydatid cyst is discussed. ? 2000 The British Infection Society PMID- 11041708 TI - Cryptococcus neoformans and cryptococcosis. PMID- 11041709 TI - Current approach to the acute management of cryptococcal infections. PMID- 11041710 TI - Failure to produce detectable antibodies to Legionella pneumophila by an immunocompetent adult. AB - A case of legionella pneumonia diagnosed by co-culture with amoebae and urinary antigen detection is described. Diagnostic antibody tests remained negative despite prolonged follow-up. Investigation showed no evidence of an under-lying immunodeficiency. The value of culture-based diagnosis and consequences of missed diagnoses are discussed. PMID- 11041711 TI - Bone marrow cryptococcal infection in the acquired immunodeficiency syndrome. AB - OBJECTIVES: To describe the bone marrow lesions in eight cases of Cryptococcus neoformans infection involving the bone marrow in HIV-infected patients. METHODS: Archival bone marrow biopsies from patients with HIV-related cryptococcosis of the bone marrow were retrospectively reviewed. Cryptocococcal organisms were identified on haematoxylin- and eosin-stained slides and confirmed using mucicarmine staining. RESULTS: Yeast cells stimulated a granulomatous response in all cases despite immunosuppression. The number of cryptococcal organisms appeared to be inversely proportional to the adequacy of the granulomatous response. All patients had a cytopenia. CONCLUSIONS: The ability to mount a tissue response in order to localize organisms is retained in patients with AIDS. Infection of the bone marrow with cryptococci may act in synergy with HIV to cause cytopenia. PMID- 11041712 TI - Treatment of a vancomycin-resistant Enterococcus faecium ventricular drain infection with quinupristin/dalfopristin and review of the literature. AB - Central nervous system infections involving vancomycin-resistant Enterococcus faecium (VREF) are infrequently described and pose significant therapeutic difficulties, because these organisms are intrinsically resistant to many antibiotics. We describe the use of intrathecal quinupristin/dalfopristin to treat a VREF-associated infection in a neuro--surgical patient. PMID- 11041714 TI - Proceedings of a workshop on micronutrients and infectious diseases: cellular and molecular immunomodulatory mechanisms. Bethesda, Maryland, USA. 16-17 September 1999. PMID- 11041713 TI - Aspergillus vertebral osteomyelitis in a child with a primary monocyte killing defect: response to GM-CSF therapy. AB - We report the first case of vertebral aspergillosis in a child with a primary defect in monocyte killing, an extremely rare immunodeficiency The diagnosis of defective monocyte killing was made by an in vitro assay that showed normal killing of Staphylococcus aureus by the patient's neutrophils but impaired killing by his monocytes. Importantly, the extensive granulomatous infection that involved the vertebral column, posterior mediastinum, pleura, and lung was not responsive to aggressive treatment with a combination of liposomal amphotericin B. intralesional amphotericin B. itraconazole, and granulocyte transfusions. Dramatic clinical and radiological improvement was only seen after the addition of granulocyte macrophage-colony stimulating factor (GM-CSF) to his treatment regimen. The use of GM-CSF in the treatment of invasive aspergillosis in immunocompromised patients requires further evaluation. PMID- 11041715 TI - Intracellular zinc depletion induces caspase activation and p21 Waf1/Cip1 cleavage in human epithelial cell lines. AB - To better understand the mechanisms by which zinc deficiency induces epithelial cell death, studies were done of the effects of intracellular zinc depletion induced by the zinc chelator TPEN on apoptosis-related events in human malignant epithelial cell lines LIM1215 (colonic), NCI-H292 (bronchial), and A549 (alveolar type II). In TPEN-treated cells, depletion of zinc was followed by activation of caspase-3 (as demonstrated by enzymatic assay and Western blotting), DNA fragmentation, and morphologic changes. Increase in caspase-3 activity began 12 h after addition of TPEN, suggesting that zinc may suppress a step just before the activation of this caspase. Caspase-6, a mediator of caspase-3 processing, also increased, but later than caspase-3. Effects of TPEN on apoptosis were completely prevented by exogenous ZnSO4 and partially prevented by peptide caspase inhibitors. A critical substrate of caspase-3 may be the cell cycle regulator p21Waf1/Cip1, which was rapidly cleaved in TPEN-treated cells to a 15-kDa fragment before further degradation. PMID- 11041716 TI - Morality, religion and metaphysics: diverse visions in bioethics. PMID- 11041717 TI - [Dacryocystorhinostomy]. PMID- 11041718 TI - Anterior introcular lens precipitates after combined phacotrabeculectomy. PMID- 11041719 TI - Silicone IOL biocompatibility--not all silicone is the same. PMID- 11041720 TI - Intraocular lens fixation. PMID- 11041721 TI - Corneal perforation during laser in situ keratomileusis. PMID- 11041722 TI - Capsule stabilization with microhooks. PMID- 11041723 TI - Surgical computerized keratometry. PMID- 11041724 TI - Mytomycin-C for post-PRK corneal haze. PMID- 11041725 TI - Retained nuclear fragments. PMID- 11041726 TI - Mini-nuc technique. PMID- 11041727 TI - [Osteo-articular ultrasonography of muscles and tendons]. PMID- 11041728 TI - [Osteo-articular ultrasonography of the shoulder]. PMID- 11041729 TI - [Ultrasonography of the wrist and the hand]. PMID- 11041730 TI - [Osteoarticular ultrasonography of the knee and the hip]. PMID- 11041731 TI - [Imaging of synovial diseases, neoplastic or non-neoplastic]. PMID- 11041732 TI - [Imaging of chronic hip pain in adults]. PMID- 11041733 TI - [Cerebellopontine angle tumors in adults]. PMID- 11041734 TI - [Neurovascular cross-compression syndromes of the cerebellopontine angle]. PMID- 11041735 TI - [Brain functional MRI: physiological, technical, and methodological bases, and clinical applications]. PMID- 11041736 TI - [Lumbar lateral recess and intervertebral foramen. Radio-anatomical study]. PMID- 11041737 TI - [Lumbar epidural space. Radio-anatomical study]. PMID- 11041738 TI - From the Centers for Disease Control and Prevention. Missed opportunities for prevention of tuberculosis among persons with HIV infection--selected locations, United States, 1996-1997. PMID- 11041739 TI - From the Centers for Disease Control and Prevention. Recall of isoniazid used for antimicrobial susceptibility testing for tuberculosis. PMID- 11041740 TI - From the Centers for Disease Control and Prevention. Update: West Nile virus activity--Northeastern United States, 2000. PMID- 11041741 TI - From the Centers for Disease Control and Prevention. Human ingestion of Bacillus anthracis-contaminated meat--Minnesota, August 2000. PMID- 11041742 TI - From the Centers for Disease Control and Prevention. Outbreak of acute febrile illness among participants in EcoChallenge Sabah 2000--Malaysia, 2000. PMID- 11041744 TI - From the Centers for Disease Control and Prevention. Progress toward global dracunculiasis eradication, June 2000. PMID- 11041743 TI - JAMA patient page. Flu vaccine. PMID- 11041745 TI - From the Centers for Disease Control and Prevention. Receipt of advice to quit smoking in Medicare managed care--United States, 1998. PMID- 11041746 TI - From the Centers for Disease Control and Prevention. Progress toward poliomyelitis eradication--African Region, 1999-March 2000. PMID- 11041748 TI - JAMA patient page. Cystic fibrosis. PMID- 11041747 TI - From the Centers for Disease Control and Prevention. Availability of influenza pandemic preparedness planning FluAid, 2.0. PMID- 11041749 TI - [Nephrology]. PMID- 11041750 TI - 13th Annual research meeting of the Kind-Philipp Foundation for Leukemia Research. 7-10 June 2000. Abstracts. PMID- 11041751 TI - [On the treatment of patients suffering from occupational diseases and those occupationally exposed]. PMID- 11041752 TI - Clopidogrel and thrombotic thrombocytopenic purpura. PMID- 11041753 TI - Clopidogrel and thrombotic thrombocytopenic purpura. PMID- 11041754 TI - Clopidogrel and thrombotic thrombocytopenic purpura. PMID- 11041755 TI - Clopidogrel and thrombotic thrombocytopenic purpura. PMID- 11041756 TI - Clopidogrel and thrombotic thrombocytopenic purpura. PMID- 11041757 TI - Observational studies and randomized trials. PMID- 11041758 TI - Observational studies and randomized trials. PMID- 11041759 TI - Observational studies and randomized trials. PMID- 11041760 TI - Observational studies and randomized trials. PMID- 11041761 TI - Observational studies and randomized trials. PMID- 11041762 TI - Effect of follicle-stimulating hormone on ovarian androgen production in a woman with isolated follicle-stimulating hormone deficiency. PMID- 11041763 TI - Anthrax: the investigation of a deadly outbreak. PMID- 11041764 TI - Aortic dissection presenting as bilateral testicular pain. PMID- 11041765 TI - Party urges Mbeki to toe the line on AIDS. PMID- 11041766 TI - [Clinical thinking and decision making in practice. A patient with persistent fever]. PMID- 11041767 TI - [Wilhelm Uhthoff: comments on cover picture]. PMID- 11041768 TI - Eletriptan in acute migraine: a double-blind, placebo-controlled comparison to sumatriptan. PMID- 11041769 TI - 30th Annual Scientific Meeting of the Australian and New Zealand Society of Nuclear Medicine. Adelaide, Australia, 20-24 May 2000. Abstracts. . PMID- 11041770 TI - Use of the World Wide Web in research: randomization in a multicenter clinical trial of treatment for twin-twin transfusion syndrome. AB - OBJECTIVE: To describe the process involved in using the World Wide Web to coordinate a randomized, multicenter international trial of treatment for twin twin transfusion syndrome. METHOD: A Web site was designed by members of the research team, a Web consultant, and a senior computer programmer. The original intent was to provide patient randomization only, but the Web site later was designed so that centers could download a data collection form. Data could be entered directly into the Web site and subsequently imported into a database at the coordinating center. EXPERIENCE: The Web site has been active for 3 years, with 13 participating centers and 31 patients enrolled. COMMENT: Use of the World Wide Web to coordinate an international, multicenter trial is an efficient method. Although there are many benefits, the most obvious is the capability to initiate and conduct a large international trial at minimal cost. PMID- 11041771 TI - The morcellator knife: a new laparoscopic instrument for supracervical hysterectomy and morcellation. PMID- 11041772 TI - [Differential therapy in primary knee joint allo-arthroplasty. Proceedings of a symposium. 4-6 March 1999, Hamburg]. PMID- 11041773 TI - Publications Bjorn Lindblom. PMID- 11041774 TI - Emergence and adaptation: studies in speech communication and language development. Proceedings of a symposium dedicated to Bjorn Lindblom onhis 65th birthday. Stockholm, 17-19 June 1999. PMID- 11041776 TI - Administration of cardioactive drugs to patients with out-of-hospital cardiac arrest. PMID- 11041775 TI - Post-radiation severe xerostomia relieved by pilocarpine: a prospective French cooperative study. AB - BACKGROUND AND PURPOSE: The aim of the study was: (1) to confirm the action of pilocarpine hydrochloride (Salagen) against xerostomia: (2) to correlate the response to dose/volume radiotherapy parameters. MATERIALS AND METHODS: From June 1995 to February 1996, 156 patients with severe radiation induced xerostomia received pilocarpine hydrochloride orally. IS mg per day with a 5 mg optional increase at S weeks up to a daily dose of 25 mg beyond 9 weeks. RESULTS: One hundred and forty five patients are fully evaluable. Treatment compliance was 75%. Thirty eight patients (26%) stopped treatment before week 12 for acute intolerance (sweating, nausea, vomiting) or no response. No severe complication occurred. Ninety ses en patients (67%) reported a significant relief of symptoms of xerostomia at 12 weeks. Within 12 weeks, the size of the subgroup ith normal food intake almost doubled (13-24 patients) while the size of the subgroup with (nearly) impossible solid food ingestion decreased by 38% (47 vs. 29 patients). The impact on quality of life was considered important or very important by 77% of the responders. CONCLUSIONS: No difference was found according to dose/volume radiotherapy parameters suggesting that oral pilocarpine hydrochloride: (1) acts primarily by stimulating minor salivary glands: (2) can be of benefit to patients suffering of severe xerostomia regardless of radiotherapy dose/volume parameters: (3) all responders are identified at 12 weeks. PMID- 11041777 TI - Genomics annotation. Beyond the first draft. PMID- 11041778 TI - Bioinformatics for biodiversity: a Web registry. PMID- 11041779 TI - Data networks. Downloading the human brain, with security. PMID- 11041780 TI - Data networks. Scientists weave new-style webs to tame the information glut. PMID- 11041781 TI - Anthropology. Misconduct alleged in Yanomamo studies. PMID- 11041782 TI - Paleoforensics. Ice Man warms up for European scientists. PMID- 11041783 TI - Genomics. Structural biology gets a $150 million boost. PMID- 11041784 TI - Presidential appointments. Panel cites barriers to government service. PMID- 11041785 TI - Neuroscience. A new look at how neurons compute. PMID- 11041787 TI - Conservation biology. Group urges southwest migration corridors. PMID- 11041786 TI - Anthropology. Bones decision rattles researchers. PMID- 11041788 TI - Marine mammalogy. Japan's whaling program carries heavy baggage. PMID- 11041789 TI - Hazardous waste cleanup. A tentative comeback for bioremediation. PMID- 11041791 TI - Many plans, one bottom line: save endangered salmon. PMID- 11041790 TI - A global biodiversity map. PMID- 11041792 TI - Many plans, one bottom line: save endangered salmon. PMID- 11041793 TI - Many plans, one bottom line: save endangered salmon. PMID- 11041794 TI - Thyroid tumor banks. PMID- 11041795 TI - Transcription. New insights into an old modification. PMID- 11041797 TI - Neuroscience. An accomplice for gamma-secretase brought into focus. PMID- 11041796 TI - Biomedicine. Protein loss in cancer cachexia. PMID- 11041798 TI - Diversity digitized. PMID- 11041799 TI - Taxonomic revival. PMID- 11041800 TI - Fossil databases move to the Web. PMID- 11041801 TI - Aspirin intolerance and related syndromes: a multidisciplinary approach. Proceedings of an international symposium. Rome, 11-13 November 1999. PMID- 11041802 TI - Influential passengers come of age. 1st International Wolbachia Conference, Orthodox Academy, Kolymbari, Crete, Greece, 7-12 June 2000. PMID- 11041803 TI - Diversity, not divergence. Genetic History of Modern Humans, 8thCEPH Annual Conference, Faculte de Medecine Lariboisiere Saint-Louis, Paris, France, 24-26 May 2000. PMID- 11041804 TI - Finding motivation at Seabrook Island: the ventral striatum, learning and plasticity. PMID- 11041805 TI - [XXIX Maghreb Medical Congress: Day hospitals. Hammamet, 7-10 May 2000. Proceedings]. PMID- 11041806 TI - Incidence of chromosome 8,10,X and Y aneuploidies in sperm nucleus of infertile men detected by FISH. AB - We studied the frequency of aneuploidy in sperm nuclei of six infertile men with abnormal semen profile and normal karyotype, using fluorescence in situ hybridization (FISH) with DNA probes for chromosomes 8, 10, X and Y. The control group consisted of four healthy fertile men with normal karyotype and semen profiles. The purpose of this study was to determine whether there are differences between infertile male donors and control donors for: (1) the incidence of sex chromosome aneuploidy, and (2) the number of disomies for chromosomes 8, and 10 cosegregating with chromosomes X and Y. FISH analysis showed no significant differences of sex ratios of the sperm nuclei in and between infertile and control groups. The most significant abnormalities in the infertile group were clusters of sperm nuclei bearing XY and XYY. In addition, the incidence of disomic sperm nuclei for chromosomes 8 and 10 consegregating with sex chromosomes was not significantly different between the patient and control groups, nor within them. However, the total frequency of aneuploid sperm nuclei was significantly different between the infertile group and the control group. We observed a significant excess of sperm nuclei bearing chromosome 10 along with disomy for chromosome Y (10YY). In conclusion, our results from FISH analysis demonstrate a significantly increased frequency of aneuploidy for the sex chromosomes in sperm nuclei from infertile men. Therefore it may be concluded that infertility is a risk factor for sex chromosome aneuploidy in sperm nuclei. PMID- 11041807 TI - [11th Workshop for Experimental and Clinical Liver Transplantation and Hepatology. Wilsede, 29 June-1 July 2000. Abstracts]. PMID- 11041808 TI - Cytology of ascitic fluid in a patient with gastric small cell carcinoma. PMID- 11041809 TI - Fine needle aspiration cytology of dermatopathic lymphadenitis. PMID- 11041810 TI - Calcifying fibrous pseudotumor: a rare entity related to inflammatory pseudotumor. PMID- 11041811 TI - Extracellular Toxoplasma organisms in granulomatous lymphadenitis. PMID- 11041812 TI - Extrathyroid variant of occult thyroid carcinoma. PMID- 11041813 TI - [Bibliography]. PMID- 11041814 TI - Schistosomiasis in the post-transmission phase. Proceedings of a meeting. San Juan, Puerto Rico. 16-17 October 1998. PMID- 11041815 TI - Pediatric intubation. PMID- 11041816 TI - A short history of a long tradition. Qi: the basis of Chinese medicine. PMID- 11041817 TI - Search for the new medicine. Patients increasingly forgo high-tech treatments for less conventional ones. PMID- 11041818 TI - Counting nurses. Data show many nursing homes to be short staffed. PMID- 11041819 TI - Short-term results of radiofrequency ablation in liver tumors. PMID- 11041820 TI - Is laparoscopic donor nephrectomy here to stay? PMID- 11041821 TI - Combined coronary artery bypass grafting and abdominal aortic aneurysm repair. PMID- 11041822 TI - Declining interest in surgical careers and the primary care mirage. PMID- 11041823 TI - [Lichen planus in the child: 25 cases. Clinical, follow-up and therapeutic aspects]. PMID- 11041824 TI - [Treatment with CO2 laser resurfacing of giant congenital nevus]. PMID- 11041825 TI - [Causative immunopathologic mechanisms in pediatric urticaria following primary injection with anti-hepatitis B vaccine]. PMID- 11041826 TI - [Eruptive pseudoangiomatosis in the adult: 8 cases]. PMID- 11041827 TI - Case report: 'treatment of malignant melanoma of the lower eyelid using anterolateral thigh flap' by T Ogawa, B Nakayama, et al. in Auris Nasus Larynx, Vol. 27;(2000):79-82. PMID- 11041828 TI - Chemistry and physics of supramolecular magnetic materials. AB - The building of multidimensional magnetic materials obtained with the molecular precursor [Cu(opba)](2-) is described. The reaction with other paramagnetic species (3d or 4f metal ions, organic radicals) yielded one-dimensional, two dimensional, and interlocked networks. The magnetic properties of these systems are reviewed using polarized neutron diffraction and magnetic measurements. It is shown that the spin density maps give a precise description of the ground state of such molecular magnetic species. Moreover, different long-range magnetic orderings (antiferro-, ferri-, and ferromagnetic) have been obtained. PMID- 11041829 TI - Do secondary orbital interactions really exist? AB - A revision of the most typical examples used to illustrate the existence of secondary orbital interactions (SOI) has been achieved. However, our analysis indicates that no conclusive evidence can be obtained from these cases. All five examples proposed by Woodward and Hoffmann in The Conservation of Orbital Symmetry have been revisited. A combination of well-known mechanisms (such as solvent effects, steric interactions, hydrogen bonds, electrostatic forces, and others) can be invoked instead to justify the endo/exo selectivity of Diels-Alder reactions. PMID- 11041830 TI - Luminescent photoproducts in UV-irradiated ice. AB - This Account describes the near-UV and visible luminescences emitted from crystalline, polycrystalline, and amorphous ices as a result of excitation by UV light. Vibrationally resolved, short-lived luminescence around 340 nm arises from excited O(2) formed by the reaction of two O atoms. Long-lived luminescence around 420 nm is tentatively assigned to a spin-forbidden (4)sigma(-) --> X(2)Pi transition of OH. This Account gives a history of the research into this little known phenomenon, places it in the context of other spectroscopic studies of gaseous and solid water, and proposes future directions for the work. PMID- 11041831 TI - Metal ion effects in isotopic hydrogen exchange in biologically important heterocycles. AB - The binding of metal ions to heteroatomic centers of biomolecules has been utilized as a probe of metal ion effects in living systems. This article focuses on the effect of N-coordination by transition metals, especially Pt(II), Co(III), Cr(III), on isotopic C(2)-H or C(8)-H exchange of imidazoles, thiazoles, and purines. The usual reactivity trend, protonated >> metalated >> neutral substrate, is excepted for Cr(III)/1-methylimidazole, where Cr(III) activates stronger than H(+). An interplay of factors is considered, including metal-to ligand back-bonding, electronic structure of metal ions, and differences in crystal field stabilization energy. PMID- 11041832 TI - Stereocontrol in cyclophane synthesis: a photochemical method to overlap aromatic rings. AB - To overlap aromatic rings in cyclophanes, the inter- and intramolecular [2 + 2] photocycloaddition of vinylarenes has been developed. It gives cyclophanes, naphthalenophanes, phenanthrenophanes, thiophenophanes, and carbazolophanes in reasonable to excellent yields. The structures and properties of cyclophanes obtained are reviewed; especially the excimer emission of phenanthrenophanes, naphthalenophanes, and carbazolophanes stressed that many intriguing cyclophanes can be obtained through the fine molecular design of precursors. PMID- 11041833 TI - Is the translational diffusion of organic radicals different from that of closed shell molecules? AB - The translational diffusion of photochemically created intermediate radicals is measured by the transient grating technique. The diffusional behavior of these intermediates is different from that of stable molecules, which have already been investigated extensively. The investigation of the diffusion of these species will provide an opportunity to reveal the unique intermolecular interaction between the intermediates and matrix. This information will be valuable for understanding photochemistry in solutions. PMID- 11041834 TI - Excited-state properties of C(60) fullerene derivatives. AB - This Account reviews our main achievements in the field of excited-state properties of fullerene derivatives. The photosensitizing and electron-acceptor features of some relevant classes of functionalized fullerene materials are highlighted, considering the impact of functionalization on fullerene characteristics. In addition, the unique optimization in terms of redox potentials, water-solubility, and singlet oxygen generation is presented for several novel fullerene-based materials. PMID- 11041835 TI - Stable silylenes. AB - The field of stable silylene research has grown dramatically since the first isolation of a stable silylene in 1994. Prior to 1994, silylenes existed only as reactive intermediates, isolable only in low-temperature matrixes. Since then, several stable silylenes have been synthesized, some in fact showing remarkable thermal stability. This Account highlights the developments in stable silylene chemistry, including theoretical and experimental studies attempting to explain the remarkable stability of the silylenes as well as the rapidly expanding reaction chemistry of the stable silylenes. PMID- 11041836 TI - Dynamic thermodynamic resolution: control of enantioselectivity through diastereomeric equilibration. AB - A theoretical foundation, tools for recognition and control, and recent examples of a class of asymmetric transformation termed dynamic thermodynamic resolution are presented. Enantioselective reaction pathways that involve an induced diastereomeric equilibration to intermediates, which are configurationally stable on the time scale of a subsequent reaction, are illustrated. Dynamic thermodynamic resolution differs from the classic, well-documented pathways of kinetic resolution and dynamic kinetic resolution in that equilibration and resolution can be operative on one system in separate controllable steps. This approach offers a high level of flexibility and provides multiple opportunities for optimization of enantioselectivity. PMID- 11041837 TI - Metal coordination and mechanism of multicopper nitrite reductase. AB - Cu-containing nitrite reductase is a homotrimer in which a ca. 36 kDa monomer contains each of type 1 Cu (two His, Cys, and Met ligands) and type 2 Cu (three His and solvent ligands). Type 1 Cu receives one electron from an electron donor and transfers it to the reaction center, type 2 Cu. The distance between the two Cu atoms bound by the Cys-His sequence segment is 12.6 A. The intramolecular electron transfer from type 1 Cu to type 2 Cu occurs probably through this segment. The noncoordinated Asp and His residues around type 2 Cu play important roles in both the electron-transfer and the catalytic processes. PMID- 11041838 TI - Identification of histidine 77 as the axial heme ligand of carbonmonoxy CooA by picosecond time-resolved resonance Raman spectroscopy. AB - The heme proximal ligand of carbonmonoxy CooA, a CO-sensing transcriptional activator, in the CO-bound form was identified to be His77 by using picosecond time-resolved resonance Raman spectroscopy. On the basis of the inverse correlation between Fe-CO and C-O stretching frequencies, we proposed previously that His77 is the axial ligand trans to CO [Uchida et al. (1998) J. Biol. Chem. 273, 19988-19992], whereas later a possibility of displacement of His77 by CO with retention of another unidentified axial ligand was reported [Vogel et al. (1999) Biochemistry 38, 2679-2687]. Although our previous resonance Raman study failed to detect the Fe-His stretching [nu(Fe-His)] mode of CO-photodissociated CooA of the carbonmonoxy adduct due to the rapid recombination, application of the picosecond time-resolved resonance Raman technique enabled us to observe a new intense line assignable to nu(Fe-His) at 211 cm(-)(1) immediately after photolysis, while it became nondiscernible after 100-ps delay. The low nu(Fe-His) frequency of photodissociated CooA indicates the presence of some strain in the Fe-His bond in CO-bound CooA. This and the rapid recombination of CO characterize the heme pocket of CooA. The 211 cm(-)(1) band was completely absent in the spectrum of the CO-photodissociated form of the His77-substituted mutant but the Fe-Im stretching band was observed in the presence of exogenous imidazole (Im). Thus, we conclude that His77 is the axial ligand of CO-bound CooA and CO displaces the axial ligand trans to His77 with retention of ligated His77 to activate CooA as the transcriptional activator. PMID- 11041839 TI - The catalytic center in nitrous oxide reductase, CuZ, is a copper-sulfide cluster. AB - The crystal structure of nitrous oxide reductase, the enzyme catalyzing the final step of bacterial denitrification in which nitrous oxide is reduced to dinitrogen, exhibits a novel catalytic site, called Cu(Z). This comprises a cluster of four copper ions bound by seven histidines and three other ligands modeled in the X-ray structure as OH(-) or H(2)O. However, elemental analyses and resonance Raman spectroscopy of isotopically labeled enzyme conclusively demonstrate that Cu(Z) has one acid-labile sulfur ligand. Thus, nitrous oxide reductase contains the first reported biological copper-sulfide cluster. PMID- 11041840 TI - Enzymatic incorporation in DNA of 1,5-anhydrohexitol nucleotides. AB - The ability of several DNA polymerases to catalyze the template-directed synthesis of duplex oligonucleotides containing a base pair between a nucleotide with anhydrohexitol ring and its natural complement has been investigated. All DNA polymerases were able to accept the chemically synthesized anhydrohexitol triphosphate as substrate and to catalyze the incorporation of one anhydrohexitol nucleotide. However, only family B DNA polymerases succeeded in elongating the primer after the incorporation of an anhydrohexitol nucleotide. In this family, Vent (exo(-)) DNA polymerase is the most successful one and was therefore selected for further investigation. Results revealed that at high enzyme concentrations six hATPs could be incorporated; however, a selective incorporation proved only feasible under experimental conditions where no more than two analogues could be inserted. Also the synthesis of a mixed HNA-DNA sequence was examined. Kinetic parameters for incorporation of one anhydrohexitol adenine nucleoside were similar to those of its natural analogue. PMID- 11041841 TI - Solution structure of the osteogenic 1-31 fragment of the human parathyroid hormone. AB - The solution conformations of a selectively osteogenic 1-31 fragment of the human parathyroid hormone (hPTH), hPTH(1-31)NH(2), have been characterized by use of very high field NMR spectroscopy at 800 MHz. The combination of the CalphaH proton and (13)Calpha chemical shifts, (3)J(NH)(alpha) coupling constants, NH proton temperature coefficients, and backbone NOEs reveals that the hPTH(1 31)NH(2) peptide has well-formed helical structures localized in two distinct segments of the polypeptide backbone. There are also many characteristic NOEs defining specific side-chain/backbone and side-chain/side-chain contacts within both helical structures. The solution structure of hPTH(1-31)NH(2) contains a short N-terminal helical segment for residues 3-11, including the helix capping residues 3 and 11 and a long C-terminal helix for residues 16-30. The two helical structures are reinforced by well-defined capping motifs and side-chain packing interactions within and at both ends of these helices. On one face of the C terminal helix, there are side-chain pairs of Glu22-Arg25, Glu22-Lys26, and Arg25 Gln29 that can form ion-pair and/or hydrogen bonding interactions. On the opposite face of this helix, there are characteristic hydrophobic interactions involving the aromatic side chain of Trp23 packing against the aliphatic side chains of Leu15, Leu24, Lys27, and Leu28. There is also a linear array of hydrophobic residues from Val2, to Leu7, to Leu11 and continuing on to residues His14 and Leu15 in the hinge region and to Trp23 in the C-terminal helix. Capping and hydrophobic interactions at the end of the N-terminal and at the beginning of the C-terminal helix appear to consolidate the helical structures into a V-shaped overall conformation for at least the folded population of the hPTH(1-31)NH(2) peptide. Stabilization of well-folded conformations in this linear 1-31 peptide fragment and possibly other analogues of human PTH may have a significant impact on the biological activities of the PTH peptides in general and specifically for the osteogenic/anabolic activities of bone-building PTH analogues. PMID- 11041842 TI - Mapping the binding of synthetic disaccharides representing epitopes of chlamydial lipopolysaccharide to antibodies with NMR. AB - A NMR study of the binding of the synthetic disaccharides alpha-Kdo-(2-->4)-alpha Kdo-(2-->O)-allyl 1 (Kdo, 3-deoxy-D-manno-oct-2-ulopyranosonic acid) and alpha Kdo-(2-->8)-alpha-Kdo-(2-->O)-allyl 2, representing partial structures of the lipopolysaccharide epitope of the intracellular bacteria Chlamydia, to corresponding monoclonal antibodies (mAbs) S23-24, S25-39, and S25-2 is presented. The conformations of 1 bound to mAbs S25-39 and of 2 bound to mAbs S23 24 and S25-39 were analyzed by employing transfer-NOESY (trNOESY) and QUIET trNOESY experiments. A quantitative analysis of QUIET-trNOESY buildup curves clearly showed that S25-39 recognized a conformation of 1 that was similar to the global energy minimum of 1, and significantly deviated from the conformation of 1 bound to mAb S25-2. For disaccharide 2, only a qualitative analysis was possible because of severe spectral overlap. Nevertheless, the analysis showed that all mAbs most likely bound to only one conformational family of 2. Saturation transfer difference (STD) NMR experiments were then employed to analyze the binding epitopes of the disaccharide ligands 1 and 2 when binding to mAbs S23-24, S25-39, and S25-2. It was found that the nonreducing pyranose unit was the major binding epitope, irrespective of the mAb and the disaccharide that were employed. Individual differences were related to the engagement of other portions of the disaccharide ligands. PMID- 11041843 TI - High pressure NMR reveals active-site hinge motion of folate-bound Escherichia coli dihydrofolate reductase. AB - A high-pressure (15)N/(1)H two-dimensional NMR study has been carried out on folate-bound dihydrofolate reductase (DHFR) from Escherichia coli in the pressure range between 30 and 2000 bar. Several cross-peaks in the (15)N/(1)H HSQC spectrum are split into two with increasing pressure, showing the presence of a second conformer in equilibrium with the first. Thermodynamic analysis of the pressure and temperature dependencies indicates that the second conformer is characterized by a smaller partial molar volume (DeltaV = -25 mL/mol at 15 degrees C) and smaller enthalpy and entropy values, suggesting that the second conformer is more open and hydrated than the first. The splittings of the cross peaks (by approximately 1 ppm on (15)N axis at 2000 bar) arise from the hinges of the M20 loop, the C-helix, and the F-helix, all of which constitute the major binding site for the cofactor NADPH, suggesting that major differences in conformation occur in the orientations of the NADPH binding units. The Gibbs free energy of the second, open conformer is 5.2 kJ/mol above that of the first at 1 bar, giving an equilibrium population of about 10%. The second, open conformer is considered to be crucial for NADPH binding, and the NMR line width indicates that the upper limit for the rate of opening is 20 s(-)(1) at 2000 bar. These experiments show that high pressure NMR is a generally useful tool for detecting and analyzing "open" structures of a protein that may be directly involved in function. PMID- 11041844 TI - Functional consequences of the Kaposi's sarcoma-associated herpesvirus protease structure: regulation of activity and dimerization by conserved structural elements. AB - The structure of Kaposi's sarcoma-associated herpesvirus protease (KSHV Pr), at 2.2 A resolution, reveals the active-site geometry and defines multiple possible target sites for drug design against a human cancer-producing virus. The catalytic triad of KSHV Pr, (Ser114, His46, and His157) and transition-state stabilization site are arranged as in other structurally characterized herpesviral proteases. The distal histidine-histidine hydrogen bond is solvent accessible, unlike the situation in other classes of serine proteases. As in all herpesviral proteases, the enzyme is active only as a weakly associated dimer (K(d) approximately 2 microM), and inactive as a monomer. Therefore, both the active site and dimer interface are potential targets for antiviral drug design. The dimer interface in KSHV Pr is compared with the interface of other herpesviral proteases. Two conserved arginines (Arg209), one from each monomer, are buried within the same region of the dimer interface. We propose that this conserved arginine may provide a destabilizing element contributing to the tuned micromolar dissociation of herpesviral protease dimers. PMID- 11041845 TI - Interhelical ion pairing in coiled coils: solution structure of a heterodimeric leucine zipper and determination of pKa values of Glu side chains. AB - Residues of opposite charge often populate heptad positions g (heptad i on chain 1) and e' (heptad i + 1 on chain 2) in dimeric coiled coils and may stabilize the dimer by formation of interchain ion pairs. To investigate the contribution to stability of such electrostatic interactions we have designed a disulfide-linked heterodimeric zipper (AB zipper) consisting of the acidic chain Ac-E-VAQLEKE VAQAEAE-NYQLEQE-VAQLEHE-CG-NH(2) and the basic chain Ac-E-VQALKKR-VQALKAR-NYAAKQK VQALRHK-CG-NH(2) in which all e and g positions are occupied by either E or K/R to form a maximum of seven interhelical salt bridges. Temperature-induced denaturation experiments monitored by circular dichroism reveal a stable coiled coil conformation below 50 degrees C and in the pH range 1.2-8.0. Stability is highest at pH approximately 4.0 [DeltaG(U) (37 degrees C) = 5.18 +/- 0.51 kcal mol(-)(1)]. The solution structure of the AB zipper at pH 5.65 has been elucidated on the basis of homonuclear (1)H NMR data collected at 800 MHz [heavy atom rmsd's for the ensemble of 50 calculated structures are 0.47 +/- 0.13 A (backbone) and 0.95 +/- 0.16 A (all)]. Both chains of the AB zipper are almost entirely in alpha-helical conformation and form a superhelix with a left-handed twist. Overhauser connectivities reveal close contacts between g position residues (heptad i on chain 1) and residues d/f (heptad i on chain 1), residues a/d (heptad i + 1 on chain 1), and residue a' (heptad i + 1 on chain 2). Residues in position e (heptad i on chain 1) are in contact with residues a/b/d/f (heptad i on chain 1) and residue d' (heptad i on chain 2). These connectivities hint at a relatively defined alignment of the side chains across the helix interface. Partial H-bond formation between the functional groups of residues g and e'(+1) is observed in the calculated structures. NMR pH titration experiments disclose pK(a) values for Glu delta-carboxylate groups: 4.14 +/- 0.02 (E(1)), 4.82 +/- 0.07 (E(6)), 4.52 +/- 0.01 (E(8)), 4.37 +/- 0.03 (E(13)), 4.11 +/- 0.02 (E(15)), 4.41 +/- 0.07 (E(20)), 4.82 +/- 0.03 (E(22)), 4.65 +/- 0.04 (E(27)), 4.63 +/- 0.03 (E(29)), 4.22 +/- 0.02 (E(1)(')). By comparison with pK(a) of Glu in unfolded peptides ( approximately 4. 3 +/- 0.1), our pK(a) data suggest marginal or even unfavorable contribution of charged Glu to the stability of the AB zipper. The electrostatic energy gained from interhelical ion pairs is likely to be surpassed by hydrophobic energy terms upon protonation of Glu, due to increased hydrophobicity of uncharged Glu and, thus, better packing against apolar residues at the chain interface. PMID- 11041846 TI - Mutant cycle analysis of the active and desensitized states of an AMPA receptor induced by willardiines. AB - The halogenated willardiines are agonists at the alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) subtype of glutamate receptors. Although they differ only by the nature of the halogen substituent, they display marked differences in their efficacy to activate the receptor channel opening and in causing desensitization. We have studied the origin of the different agonist properties of the willardiines and in particular the nature of the structural element within the receptor binding domain that is able to distinguish between willardiines at a subatomic resolution of 0.6 A (the difference in radius between F and Br) and allow (S)-5-fluorowillardiine to cause receptor desensitization much more than (S)-5-bromowillardiine. For this purpose, we analyzed, with the thermodynamic mutant cycle method, the active and desensitized states induced by the willardiines in the GluR1 subtype of AMPA receptors and GluR1 mutants in which residues E398, Y446, L646, and S650, within the agonist binding domain, were mutated. The results were used to generate a 3D model of the willardiine docking mode. We suggest that the active and desensitized states of the AMPA-R correspond, respectively, to the open-lobe and closed-lobe conformations of the agonist binding domain. PMID- 11041847 TI - Arrestin isoforms dictate differential kinetics of A2B adenosine receptor trafficking. AB - Adenosine mediates the activation of adenylyl cyclase via its interaction with specific A(2A) and A(2B) adenosine receptors. Previously, we demonstrated that arrestins are involved in rapid agonist-promoted desensitization of the A(2B) adenosine receptor (A(2B)AR) in HEK293 cells. In the present study, we investigate the role of arrestins in A(2B)AR trafficking. Initial studies demonstrated that HEK293 cells stably expressing arrestin antisense constructs, which reduce endogenous arrestin levels, effectively reduced A(2B)AR internalization. A(2B)AR recycling after agonist-induced endocytosis was also significantly impaired in cells with reduced arrestin levels. Interestingly, while overexpression of arrestin-2 or arrestin-3 rescued A(2B)AR internalization and recycling, arrestin-3 promoted a significantly faster rate of recycling as compared to arrestin-2. The specificity of arrestin interaction with A(2B)ARs was further investigated using arrestins fused to the green fluorescent protein (arr 2-GFP and arr-3-GFP). Both arrestins underwent rapid translocation (<1 min) from the cytosol to the plasma membrane following A(2B)AR activation. However, longer incubations with agonist (>10 min) revealed that arr-2-GFP but not arr-3-GFP colocalized with the A(2B)AR in rab-5 and transferrin receptor containing early endosomes. At later times, the A(2B)AR but not arr-2-GFP was observed in an apparent endocytic recycling compartment. Thus, while arrestin-2 and arrestin-3 mediate agonist-induced A(2B)AR internalization with relative equal potency, arrestin isoform binding dictates the differential kinetics of A(2B)AR recycling and resensitization. PMID- 11041848 TI - Comparison of the structure of vMIP-II with eotaxin-1, RANTES, and MCP-3 suggests a unique mechanism for CCR3 activation. AB - Herpesvirus-8 macrophage inflammatory protein-II (vMIP-II) binds a uniquely wide spectrum of chemokine receptors. We report the X-ray structure of vMIP-II determined to 2.1 A resolution. Like RANTES, vMIP-II crystallizes as a dimer and displays the conventional chemokine tertiary fold. We have compared the surface topology and electrostatic potential of vMIP-II to those of eotaxin-1, RANTES, and MCP-3, three CCR3 physiological agonists with known three-dimensional structures. Surface epitopes identified on RANTES to be involved in binding to CCR3 are mimicked on the eotaxin-1 and MCP-3 surface. However, the surface topology of vMIP-II in these regions is markedly different. The results presented here indicate that the structural basis for interaction with the chemokine receptor CCR3 by vMIP-II is different from that for the physiological agonists eotaxin-1, RANTES, and MCP-3. These differences on vMIP-II may be a consequence of its broad-range receptor recognition capabilities. PMID- 11041849 TI - Structures of the contryphan family of cyclic peptides. Role of electrostatic interactions in cis-trans isomerism. AB - The contryphan family of cyclic peptides, isolated recently from various species of cone shell, has the conserved sequence motif NH(3)(+)-X(1)COD-WX(5)PWC-NH(2), where X(1) is either Gly or absent, O is 4-trans-hydroxyproline, and X(5) is Glu, Asp, or Gln. The solution structures described herein of two new naturally occurring contryphan sequences, contryphan-Sm and des[Gly1]-contryphan-R, are similar to those of contryphan-R, the structure of which has been determined recently [Pallaghy et al. (1999) Biochemistry 38, 11553-11559]. The (1)H NMR chemical shifts of another naturally occurring peptide, contryphan-P, indicate that it also adopts a similar structure. All of these contryphans exist in solution as a mixture of two conformers due to cis-trans isomerization about the Cys2-Hyp3 peptide bond. The lower cis-trans ratio for contryphan-Sm enabled elucidation of the 3D structure of both its major and its minor forms, for which the patterns of (3)J(H)(alpha)(HN) coupling constants are very different. As with contryphan-R, the structure of the major form of contryphan-Sm (cis Cys2-Hyp3 peptide bond) contains an N-terminal chain reversal and a C-terminal type I beta turn. The minor conformer (trans peptide bond) forms a hairpin structure with sheetlike hydrogen bonds and a type II beta-turn, with the D-Trp4 at the 'Gly position' of the turn. The ratio of conformers arising from cis-trans isomerism around the peptide bond preceding Hyp3 is sensitive to both the amino acid sequence and the solution conditions, varying from 2.7:1 to 17:1 across the five sequences. The sequence and structural determinants of the cis-trans isomerism have been elucidated by comparison of the cis-trans ratios for these peptides with those for contryphan-R and an N-acetylated derivative thereof. The cis-trans ratio is reduced for peptides in which either the charged N-terminal ammonium or the X(5) side-chain carboxylate is neutralized, implying that an electrostatic interaction between these groups stabilizes the cis conformer relative to the trans. These results on the structures and cis-trans equilibrium of different conformers suggest a paradigm of 'locally determined but globally selected' folding for cyclic peptides and constrained protein loops, where the series of stereochemical centers in the loop dictates the favorable conformations and the equilibrium is determined by a small number of side-chain interactions. PMID- 11041850 TI - Structural studies of lysyl-tRNA synthetase: conformational changes induced by substrate binding. AB - Lysyl-tRNA synthetase is a member of the class II aminoacyl-tRNA synthetases and catalyses the specific aminoacylation of tRNA(Lys). The crystal structure of the constitutive lysyl-tRNA synthetase (LysS) from Escherichia coli has been determined to 2.7 A resolution in the unliganded form and in a complex with the lysine substrate. A comparison between the unliganded and lysine-bound structures reveals major conformational changes upon lysine binding. The lysine substrate is involved in a network of hydrogen bonds. Two of these interactions, one between the alpha-amino group and the carbonyl oxygen of Gly 216 and the other between the carboxylate group and the side chain of Arg 262, trigger a subtle and complicated reorganization of the active site, involving the ordering of two loops (residues 215-217 and 444-455), a change in conformation of residues 393 409, and a rotation of a 4-helix bundle domain (located between motif 2 and 3) by 10 degrees. The result of these changes is a closing up of the active site upon lysine binding. PMID- 11041851 TI - Role of SRP19 in assembly of the Archaeoglobus fulgidus signal recognition particle. AB - Previous studies have shown that SRP19 promotes association of the highly conserved signal peptide-binding protein, SRP54, with the signal recognition particle (SRP) RNA in both archaeal and eukaryotic model systems. In vitro characterization of this process is now reported using recombinantly expressed components of SRP from the hyperthermophilic, sulfate-reducing archaeon Archaeoglobus fulgidis. A combination of native gel mobility shift, filter binding, and Ni-NTA agarose bead binding assays were used to determine the binding constants for binary and ternary complexes of SRP proteins and SRP RNA. Archaeal SRP54, unlike eukaryotic homologues, has significant intrinsic affinity for 7S RNA (K(D) approximately 15 nM), making it possible to directly compare particles formed in the presence and absence of SRP19 and thereby assess the precise role of SRP19 in the assembly process. Chemical modification studies using hydroxyl radicals and DEPC identify nonoverlapping primary binding sites for SRP19 and SRP54 corresponding to the tips of helix 6 and helix 8 (SRP19) and the distal loop and asymmetric bulge of helix 8 (SRP54). SRP19 additionally induces conformational changes concentrated in the proximal asymmetric bulge of helix 8. Selected nucleotides in this bulge become modified as a result of SRP19 binding but are subsequently protected from modification by formation of the complete complex with SRP54. Together these results suggest a model for assembly in which bridging the ends of helix 6 and helix 8 by SRP19 induces a long-range structural change to present the proximal bulge in a conformation compatible with high-affinity SRP54 binding. PMID- 11041852 TI - Double-stranded RNA adenosine deaminases ADAR1 and ADAR2 have overlapping specificities. AB - Adenosine deaminases that act on RNA (ADARs) deaminate adenosines to produce inosines within RNAs that are largely double-stranded (ds). Like most dsRNA binding proteins, the enzymes will bind to any dsRNA without apparent sequence specificity. However, once bound, ADARs deaminate certain adenosines more efficiently than others. Most of what is known about the intrinsic deamination specificity of ADARs derives from analyses of Xenopus ADAR1. In addition to ADAR1, mammalian cells have a second ADAR, named ADAR2; the deamination specificity of this enzyme has not been rigorously studied. Here we directly compare the specificity of human ADAR1 and ADAR2. We find that, like ADAR1, ADAR2 has a 5' neighbor preference (A approximately U > C = G), but, unlike ADAR1, also has a 3' neighbor preference (U = G > C = A). Simultaneous analysis of both neighbor preferences reveals that ADAR2 prefers certain trinucleotide sequences (UAU, AAG, UAG, AAU). In addition to characterizing ADAR2 preferences, we analyzed the fraction of adenosines deaminated in a given RNA at complete reaction, or the enzyme's selectivity. We find that ADAR1 and ADAR2 deaminate a given RNA with the same selectivity, and this appears to be dictated by features of the RNA substrate. Finally, we observed that Xenopus and human ADAR1 deaminate the same adenosines on all RNAs tested, emphasizing the similarity of ADAR1 in these two species. Our data add substantially to the understanding of ADAR2 specificity, and aid in efforts to predict which ADAR deaminates a given editing site adenosine in vivo. PMID- 11041853 TI - Structural basis for substrate specificity differences of horse liver alcohol dehydrogenase isozymes. AB - A structure determination in combination with a kinetic study of the steroid converting isozyme of horse liver alcohol dehydrogenase, SS-ADH, is presented. Kinetic parameters for the substrates, 5beta-androstane-3beta,17beta-ol, 5beta androstane-17beta-ol-3-one, ethanol, and various secondary alcohols and the corresponding ketones are compared for the SS- and EE-isozymes which differ by nine amino acid substitutions and one deletion. Differences in substrate specificity and stereoselectivity are explained on the basis of individual kinetic rate constants for the underlying ordered bi-bi mechanism. SS-ADH was crystallized in complex with 3alpha,7alpha,12alpha-trihydroxy-5beta-cholan -24 acid (cholic acid) and NAD(+), but microspectrophotometric analysis of single crystals proved it to be a mixed complex containing 60-70% NAD(+) and 30-40% NADH. The crystals belong to the space group P2(1) with cell dimensions a = 55.0 A, b = 73.2 A, c = 92.5 A, and beta = 102.5 degrees. A 98% complete data set to 1.54-A resolution was collected at 100 K using synchrotron radiation. The structure was solved by the molecular replacement method utilizing EE-ADH as the search model. The major structural difference between the isozymes is a widening of the substrate channel. The largest shifts in C(alpha) carbon positions (about 5 A) are observed in the loop region, in which a deletion of Asp115 is found in the SS isozyme. SS-ADH easily accommodates cholic acid, whereas steroid substrates of similar bulkiness would not fit into the EE-ADH substrate site. In the ternary complex with NAD(+)/NADH, we find that the carboxyl group of cholic acid ligates to the active site zinc ion, which probably contributes to the strong binding in the ternary NAD(+) complex. PMID- 11041854 TI - Optimization of the P'-region of peptide inhibitors of hepatitis C virus NS3/4A protease. AB - Infection by Hepatitis C Virus (HCV) leads to a slowly progressing disease that over two decades can lead to liver cirrhosis or liver cancer. Currently, one of the most promising approaches to anti-HCV therapy is the development of inhibitors of the NS3/4A protease, which is essential for maturation of the viral polyprotein. Several substrate-derived inhibitors of NS3/4A have been described, all taking advantage of binding to the S subsite of the enzyme. Inspection of the S' subsite of NS3/4A shows binding pockets which might be exploited for inhibitor binding, but due to the fact that ground-state binding to the S' subsite is not used by the substrate, this does not represent a suitable starting point. We have now optimized S'-binding in the context of noncleavable decapeptides spanning P6 P4'. Binding was sequentially increased by introduction of the previously optimized P-region [Ingallinella et al. (1998) Biochemistry 37, 8906-8914], change of the P4' residue, and combinatorial optimization of positions P2'-P3'. The overall process led to an increase in binding of more than 3 orders of magnitude, with the best decapeptide showing IC(50) < 200 pM. The binding mode of the decapeptides described in the present work shares features with the binding mode of the natural substrates, together with novel interactions within the S' subsite. Therefore, these peptides may represent an entry point for a novel class of NS3 inhibitors. PMID- 11041855 TI - Conformational changes in photosystem II supercomplexes upon removal of extrinsic subunits. AB - Photosystem II is a multisubunit pigment-protein complex embedded in the thylakoid membranes of chloroplasts. It consists of a large number of intrinsic membrane proteins involved in light-harvesting and electron-transfer processes and of a number of extrinsic proteins required to stabilize photosynthetic oxygen evolution. We studied the structure of dimeric supercomplexes of photosystem II and its associated light-harvesting antenna by electron microscopy and single particle image analysis. Comparison of averaged projections from native complexes and complexes without extrinsic polypeptides indicates that the removal of 17 and 23 kDa extrinsic subunits induces a shift of about 1.2 nm in the position of the monomeric peripheral antenna protein CP29 toward the central part of the supercomplex. Removal of the 33 kDa extrinsic protein induces an inward shift of the strongly bound trimeric light-harvesting complex II (S-LHCII) of about 0.9 nm, and in addition destabilizes the monomer-monomer interactions in the central core dimer, leading to structural rearrangements of the core monomers. It is concluded that the extrinsic subunits keep the S-LHCII and CP29 subunits in proper positions at some distance from the central part of the photosystem II core dimer to ensure a directed transfer of excitation energy through the monomeric peripheral antenna proteins CP26 and CP29 and/or to maintain sequestered domains of inorganic cofactors required for oxygen evolution. PMID- 11041856 TI - CD39L2, a gene encoding a human nucleoside diphosphatase, predominantly expressed in the heart. AB - E-NTPDases are extracellular enzymes that hydrolyze nucleotides. The human E NTPDase gene family currently consists of five reported members (CD39, CD39L1, CD39L2, CD39L3, and CD39L4). Both membrane-bound and secreted family members have been predicted by encoded transmembrane and leader peptide motifs. In this report, we demonstrate that the human CD39L2 gene is expressed predominantly in the heart. In situ hybridization results from heart indicate that the CD39L2 message is expressed in muscle and capillary endothelial cells. We also show that the CD39L2 gene encodes an extracellular E-NTPDase. Flow cytometric experiments show that transiently expressed CD39L2 is present on the surface of COS-7 cells. Transfected cells also produce recombinant glycosylated protein in the medium, and this process can be blocked by brefeldin A, an inhibitor of the mammalian secretory pathway. The enzymology of CD39L2 shows characteristic features of a typical E-NTPDase, but with a much higher degree of specificity for NDPs over NTPs as enzymatic substrates. The kinetics of the ADPase activity exhibit positive cooperativity. The predominance of CD39L2 expression in the heart supports a functional role in regulating platelet activation and recruitment in this organ. PMID- 11041857 TI - Biochemical characterization of CD39L4. AB - Nucleotides are involved in regulating a number of important processes ranging from inflammation to platelet aggregation. Enzymes that can modulate levels of nucleotides in the blood therefore represent important regulatory components in these physiological systems. CD39L4 is a soluble E-nucleoside triphosphate dephosphohydrolase (E-NTPDase) with specificity for nucleotide diphosphates (NDPs). In this study, stable mammalian and insect cell lines were generated expressing CD39L4 protein to purify and characterize the recombinant protein. We demonstrate that recombinant CD39L4 protein expressed in human embryonic carcinoma 293 cells is glycosylated by comparing the molecular masses before and after glycosidase treatment. Activity measurements of CD39L4 isolated from tunicamycin-treated, transiently transfected COS-7 cells indicate that glycosylation is not required for full ADPase activity. Recombinant human CD39L4 protein isolated from stable insect cells was glycosylated differently, but also demonstrated relative activity comparable to that of the mammalian protein. When denatured by SDS under nonreducing conditions, a fraction of the CD39L4 protein migrates as a 110 kDa disulfide-linked dimer. We determined that the monomer is the most active form of CD39L4 by measuring the activity of sucrose density gradient fractions of monomers and partially purified dimers. The physiological significance of the biochemical and enzymatic characterization is discussed. PMID- 11041858 TI - Phosphorylation of serine 51 in initiation factor 2 alpha (eIF2 alpha) promotes complex formation between eIF2 alpha(P) and eIF2B and causes inhibition in the guanine nucleotide exchange activity of eIF2B. AB - Phosphorylation of serine 51 residue on the alpha-subunit of eukaryotic initiation factor 2 (eIF2alpha) inhibits the guanine nucleotide exchange (GNE) activity of eIF2B, presumably, by forming a tight complex with eIF2B. Inhibition of the GNE activity of eIF2B leads to impairment in eIF2 recycling and protein synthesis. We have partially purified the wild-type (wt) and mutants of eIF2alpha in which the serine 51 residue was replaced with alanine (51A mutant) or aspartic acid (51D mutant) in the baculovirus system. Analysis of these mutants has provided novel insight into the role of 51 serine in the interaction between eIF2 and eIF2B. Neither mutant was phosphorylated in vitro. Both mutants decreased eIF2alpha phosphorylation occurring in hemin and poly(IC)-treated reticulocyte lysates due to the activation of double-stranded RNA-dependent protein kinase (PKR). However, addition of 51D, but not 51A mutant eIF2alpha protein promoted inhibition of the GNE activity of eIF2B in hemin-supplemented rabbit reticulocyte lysates in which relatively little or no endogenous eIF2alpha phosphorylation occurred. The 51D mutant enhanced the inhibition in GNE activity of eIF2B that occurred in hemin and poly(IC)-treated reticulocyte lysates where PKR is active. Our results show that the increased interaction between eIF2 and eIF2B protein, occurring in reticulocyte lysates due to increased eIF2alpha phosphorylation, is decreased significantly by the addition of mutant 51A protein but not 51D. Consistent with the idea that mutant 51D protein behaves like a phosphorylated eIF2alpha, addition of this partially purified recombinant subunit, but not 51A or wt eIF2alpha, increases the interaction between eIF2 and 2B proteins in actively translating hemin-supplemented lysates. These findings support the idea that phosphorylation of the serine 51 residue in eIF2alpha promotes complex formation between eIF2alpha(P) and eIF2B and thereby inhibits the GNE activity of eIF2B. PMID- 11041859 TI - Kinetic characterization of the second step of group II intron splicing: role of metal ions and the cleavage site 2'-OH in catalysis. AB - The ai5gamma group II intron from yeast excises itself from precursor transcripts in the absence of proteins. When a shortened form of the intron containing all but the 3'-terminal six nucleotides is incubated with an exon 1 oligonucleotide and a 3' splice site oligonucleotide, a nucleotidyl transfer reaction occurs that mimics the second step of splicing. As this tripartite reaction provides a means to identify important functional groups in 3' splice site recognition and catalysis, we establish here a minimal kinetic framework and demonstrate that the chemical step is rate-limiting. We use this framework to characterize the metal ion specificity switch observed previously upon sulfur substitution of the 3' oxygen leaving group and to elucidate by atomic mutagenesis the role of the neighboring 2'-OH in catalysis. The results suggest that both the 3'-oxygen leaving group and the neighboring 2'-OH are important ligands for metal ions in the transition state but not in the ground state and that the 2'-OH may play an additional role in transition state stabilization by donating a hydrogen bond. Metal specificity switch experiments combined with quantitative analysis show that the Mn(2+) that interacts with the leaving group binds to the ribozyme with the same affinity as the metal ion that interacts with the neighboring 2'-OH, raising the possibility that a single metal ion mediates interactions with the 2' and 3'-oxygen atoms at the 3' splice site. PMID- 11041860 TI - Functional characterization of zinc-finger motif in redox regulation of RPA-ssDNA interaction. AB - The 70-kDa subunit of eukaryotic replication protein A (RPA) contains a conserved four cysteine-type zinc-finger motif that has been implicated in regulation of DNA replication and repair. Unlike other zinc-finger proteins, RPA zinc-finger motif is not a DNA-binding component, and deletion of the zinc-finger had very little effect on its ssDNA binding activity. Recently, we described a novel function for the zinc-finger motif in regulation of RPA's ssDNA binding activity through reduction-oxidation (redox). In this study, we carried out a detailed analysis of wild-type RPA and zinc-finger mutants in redox regulation of their ssDNA binding activity. Any mutation at a zinc-finger cysteine abolished its redox role in regulation of RPA-ssDNA interaction, suggesting that all four zinc finger cysteines are required for redox regulation. Reactivity of cysteine residues to 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) indicated that wild-type RPA contained 8.2 reactive thiols/molecule including all four cysteines in the zinc-finger motif. Zinc-finger cysteines slowly reacted to DTNB as compared to others. Zn(II) was not only essential but also uniquely qualified for redox regulation of RPA-ssDNA interaction, suggesting that Zn(II)-cysteine coordination is crucial for the zinc-finger function. Redox status significantly affected initial interaction of RPA with ssDNA but had no effect after RPA formed a stable complex with DNA. Together, our results suggest that the zinc-finger motif mediates the transition of RPA-ssDNA interaction to a stable RPA-ssDNA complex in a redox-dependent manner. PMID- 11041862 TI - Helical junctions as determinants for RNA folding: origin of tertiary structure stability of the hairpin ribozyme. AB - Helical junctions are ubiquitous structural elements that govern the folding and tertiary structure of RNAs. The tobacco ringspot virus hairpin ribozyme consists of two helix-loop-helix elements that lie on adjacent arms of a four-way junction. In the active form of the hairpin ribozyme, the loops are in proximity. The nature of the helical junction determines the stability of the hairpin ribozyme tertiary structure [Walter, N. G., Burke, J. M., and Millar, D. P. (1999) Nat. Struct. Biol. 6, 544-549] and thus its catalytic activity. We used two-, three-, and four-way junction hairpin ribozymes as model systems to investigate the thermodynamic basis for the different tertiary structure stabilities. The equilibrium between docked and extended conformers was analyzed as a function of temperature using time-resolved fluorescence resonance energy transfer (trFRET). As the secondary and tertiary structure transitions overlap, information from UV melting curves and trFRET had to be combined to gain insight into the thermodynamics of both structural transitions. It turned out that the higher tertiary structure stability observed in the context of a four-way junction is the result of a lower entropic cost for the docking process. In the two- and three-way junction ribozymes, a high entropic cost counteracts the favorable enthalpic term, rendering the docked conformer only marginally stable. Thus, two- and three-way junction tertiary structures are more sensitive toward regulation by ligands, whereas four-way junctions provide a stable scaffold. Altogether, RNA folding and stability appear to be governed by principles similar to those for the folding of proteins. PMID- 11041861 TI - Interactions of nucleotide cofactors with the Escherichia coli replication factor DnaC protein. AB - Quantitative analyses of the interactions of nucleotide cofactors with the Escherichia coli replicative factor DnaC protein have been performed using thermodynamically rigorous fluorescence titration techniques. This approach allowed us to obtain stoichiometries of the formed complexes and interaction parameters, without any assumptions about the relationship between the observed signal and the degree of binding. The stoichiometry of the DnaC-nucleotide complex has been determined in direct binding experiments with fluorescent nucleotide analogues, MANT-ATP and MANT-ADP. The stoichiometry of the DnaC complexes with unmodified ATP and ADP has been determined using the macromolecular competition titration method (MCT). The obtained results established that at saturation the DnaC protein binds a single nucleotide molecule per protein monomer. Analyses of the binding of fluorescent analogues and unmodified nucleotides to the DnaC protein show that ATP and ADP have the same affinities for the nucleotide-binding site, albeit the corresponding complexes have different structures, specifically affected by the presence of magnesium cations in solution. Although the presence of the gamma-phosphate does not affect the affinity, the structure of the triphosphate group is critical. While the affinity of ATP-gamma-S is the same as the affinity of ATP, the affinities of AMP-PNP and AMP-PCP are approximately 2 and approximately 4 orders lower than that of ATP, respectively. Moreover, the ribose plays a significant role in forming a stable complex. The binding constants of dATP and dADP are approximately 2 orders of magnitude lower than those for ribose nucleotides. The nucleotide-binding site of the DnaC protein is highly base specific. The intrinsic affinity of adenosine triphosphates and diphosphates is at least 3-4 orders of magnitude higher than for any of the other examined nucleotides. The obtained data indicate that the recognition mechanism of the nucleotide by the structural elements of the binding site is complex with the base providing the specificity and the ribose, as well as the second phosphate group contributing to the affinity. The significance of the results for the functioning of the DnaC protein is discussed. PMID- 11041863 TI - Functional hydrogen-bonding map of the minor groove binding tracks of six DNA polymerases. AB - Recent studies have identified amino acid side chains forming several hydrogen bonds in the DNA minor groove as potentially important in polymerase replication of DNA. Few studies have probed these interactions on the DNA itself. Using non hydrogen-bonding nucleoside isosteres, we have now studied effects in both primer and template strands with several polymerases to investigate the general importance of these interactions. All six polymerases show differences in the H bonding effects in the minor groove. Two broad classes of activity are seen, with a first group of DNA polymerases (KF(-), Taq, and HIV-RT) that efficiently extends nonpolar base pairs containing nucleoside Q (9-methyl-1H-imidazo[4,5 b]pyridine) but not the analogue Z (4-methylbenzimidazole), implicating a specific minor groove interaction at the first extension site. A second group of polymerases (Pol alpha, Pol beta, and T7(-)) fails to extend all non-H-bonding base pairs, indicating that these enzymes may need minor groove hydrogen bonds at both minor groove sites or that they are especially sensitive to noncanonical DNA structure or stability. All DNA polymerases examined use energetically important minor groove interactions to probe newly synthesized base pairs before extending them. The positions of these interactions vary among the enzymes, and only a subset of the interactions identified structurally appears to be functionally important. In addition, polymerases appear to be differently sensitive to small changes in base pair geometry. PMID- 11041864 TI - Identification of the structural subunits required for formation of the metal centers in subunit I of cytochrome c oxidase of Rhodobacter sphaeroides. AB - Genetic manipulation of the aa(3)-type cytochrome c oxidase of Rhodobacter sphaeroides was used to determine the minimal structural subunit associations required for the assembly of the heme A and copper centers of subunit I. In the absence of the genes for subunits II and III, expression of the gene for subunit I in Rb. sphaeroides allowed purification of a form of free subunit I (subunit I(a)()) that contained a single heme A. No copper was present in this protein, indicating that the heme a(3)-Cu(B) active site was not assembled. In cells expressing the genes for subunits I and II, but not subunit III, two oxidase forms were synthesized that were copurified by histidine affinity chromatography and separated by anion-exchange chromatography. One form was a highly active subunit I-II oxidase containing a full complement of structurally normal metal centers. This shows that association of subunit II with subunit I is required for stable formation of the active site in subunit I. In contrast, subunit III is not required for the formation of any of the metal centers or for the production of an oxidase with wild-type activity. The second product of the cells lacking subunit III was a large amount of a free form of subunit I that appeared identical to subunit I(a)(). Since significant amounts of subunit I(a)() were also isolated from wild-type cells, it is likely that subunit I(a)() will be present in any preparation of the aa(3)-type oxidase isolated via an affinity tag on subunit I. PMID- 11041865 TI - Detergent-solubilized bovine cytochrome c oxidase: dimerization depends on the amphiphilic environment. AB - The extent to which bovine cytochrome c oxidase (COX) dimerizes in nondenaturing detergent environments was assessed by sedimentation velocity and equilibrium. In contrast to generally accepted opinion, the COX dimer is difficult to maintain and is the major oligomeric form only when COX is solubilized with a low concentration of dodecylmaltoside, i.e., approximately 1 mg/mg protein. The dimer form is intrinsically unstable and dissociates into monomers with increased detergent concentration, i.e., >5 mg/mg protein. The structure of the solubilizing detergent, however, greatly alters detergent effectiveness by inducing either monomerization or aggregation. Triton X-100 is most effective at solubilizing COX, but it destabilizes COX dimers, even at low concentration. Undecylmaltoside, decylmaltoside, and octaethyleneglycolmonododecyl ether (C(12)E(8)) are less effective at solubilizing COX. Each prevents COX aggregation at high detergent concentration, but also destabilizes the COX dimer. Other detergents, e.g., Tween 20, sodium cholate, sodium deoxycholate, CHAPS, or CHAPSO, are completely ineffective COX solubilizers and do not prevent aggregation even at 10-40 mg/mL. The transition from dimers to monomers depends on many factors other than detergent structure and concentration, e.g., protein concentration, phospholipid content and pH. We conclude that the intrinsic dimeric structure of COX can be maintained only after solubilization with low concentrations of dodecylmaltoside at near neutral pH, and even then precautions must be taken to prevent its dissociation into monomers. PMID- 11041866 TI - The quantitatively important relationship between homocysteine metabolism and glutathione synthesis by the transsulfuration pathway and its regulation by redox changes. AB - Homocysteine is a key junction metabolite in methionine metabolism. It suffers two major metabolic fates: transmethylation catalyzed by methionine synthase or betaine homocysteine methyl transferase and transsulfuration catalyzed by cystathionine beta-synthase leading to cystathionine. The latter is subsequently converted to cysteine, a precursor of glutathione. Studies with purified mammalian methionine synthase and cystathionine beta-synthase have revealed the oxidative sensitivity of both junction enzymes, suggesting the hypothesis that redox regulation of this pathway may be physiologically significant. This hypothesis has been tested in a human hepatoma cell line in culture in which the flux of homocysteine through transsulfuration under normoxic and oxidative conditions has been examined. Addition of 100 microM H(2)O(2) or tertiary butyl hydroperoxide increased cystathionine production 1.6- and 2.1-fold from 82 +/- 7 micromol h(-)(1) (L of cells)(-)(1) to 136 +/- 15 and 172 +/- 23 micromol h(-)(1) (L of cells)(-)(1), respectively. The increase in homocysteine flux through the transsulfuration pathway exhibited a linear dose dependence on the concentrations of both oxidants (50-200 microM H(2)O(2) and 10-200 microM tertiary butyl hydroperoxide). Furthermore, our results reveal that approximately half of the intracellular glutathione pool in human liver cells is derived from homocysteine via the transsulfuration pathway. The redox sensitivity of the transsulfuration pathway can be rationalized as an autocorrective response that leads to an increased level of glutathione synthesis in cells challenged by oxidative stress. In summary, this study demonstrates the importance of the homocysteine-dependent transsulfuration pathway in the maintenance of the intracellular glutathione pool, and the regulation of this pathway under oxidative stress conditions. Aberrations in this pathway could compromise the redox buffering capacity of cells, which may in turn be related to the pathophysiology of the different homocysteine-related diseases. PMID- 11041867 TI - Influence of the axial ligands on the spectral properties of P700 of photosystem I: a study of site-directed mutants. AB - Two histidines provide the axial ligands of the two chlorophyll a (Chl a) molecules which form the primary electron donor (P700) of photosystem I (PSI). Histidine 676 in the protein subunit PsaA, His(A676), and histidine 656 in subunit PsaB, His(B656), were replaced in the green algae Chlamydomnas reinhardtii by site-directed mutagenesis with nonpolar, uncharged polar, acidic, and basic amino acid residues. Only the substitutions with uncharged polar residues led to a significant accumulation of PSI in the thylakoid membranes. These PSI complexes were isolated and the physical properties of the primary donor characterized. The midpoint potential of P700(+)(*)/P700 was increased in all mutants (up to 140 mV) and showed a dependence on size and polarizability of the residues when His(B656) was substituted. In the light-minus-dark absorbance spectra, all mutations in PsaB exhibited an additional bleaching band at 665 nm at room temperature comparable with the published spectrum for the replacement of His(B656) with asparagine [Webber, A. N., Su Hui, Bingham, S. E., Kass, H., Krabben, L., Kuhn, M., Jordan, R., Schlodder, E., and Lubitz, W. (1996) Biochemistry 35, 12857-12863]. Substitutions of His(A676) showed an additional shoulder around 680 nm. In the low-temperature absorbance difference spectra of P700(+)(*)/P700, a blue shift of the main bleaching band by 2 nm and some changes in the spectral features around 660 nm were observed for mutations of His(B656) in PsaB. The analogous substitution in PsaA showed only a shift of the main bleaching band. Similar effects of the mutations were found in the (3)P700/P700 absorbance difference spectra at low temperatures (T = 2 K). The zero-field splitting parameters of (3)P700 were not significantly changed in the mutated PSI complexes. The electron spin density distribution of P700(+)(*), determined by ENDOR spectroscopy, was only changed when His(B656) was replaced. In all measurements, two general observations were made. (i) The replacement of His(B656) had a much stronger impact on the physical properties of P700 than the mutation of His(A676). (ii) The exchange of His(B656) with glutamine induces the smallest changes in the spectra or the midpoint potential, whereas the other replacements exhibited a stronger but very similar influence on the spectroscopic features of P700. The data provide convincing evidence that the unpaired electron in the cation radical and the triplet state of P700 are mainly localized on the Chl a of the dimer which is axially coordinated by His(B656). PMID- 11041868 TI - Multidrug resistance protein MRP1 reconstituted into lipid vesicles: secondary structure and nucleotide-induced tertiary structure changes. AB - Multidrug resistance protein MRP1 is an ATP-dependent drug efflux pump that confers resistance in human cancer cells to various chemotherapeutic drugs. We have reconstituted purified MRP1 in lipid vesicles. The reconstituted protein conserves ATPase and drug transport activity. Structural analysis of MRP1 was investigated by infrared spectroscopy for the first time. This technique offers a unique opportunity to determine structural parameters characterizing a membrane protein in its lipid environment. Addition of different ligands (MgATP, MgATPgammaS, MgADP and P(i), and MgADP) did not significantly affect the MRP1 secondary structure, which is made of 46% alpha-helix, 26% beta-sheet, 12% beta turns, and 17% random coil. Binding of MgATP increased the protein accessibility to the solvent, suggesting a modification in the tertiary organization of the protein. Hydrolysis of MgATP to MgADP and P(i) did not significantly change the global accessibility of the protein. Release of P(i), after hydrolysis, caused a decrease in the accessibility of MRP1 to the water phase which brings the protein back to its initial conformation. All together, the data demonstrate that MRP1 adopts different structures during its catalytic cycle. PMID- 11041869 TI - Characterization of human copine III as a phosphoprotein with associated kinase activity. AB - The copines, first described by Creutz et al. [(1998) J. Biol. Chem. 273, 1393 1402], comprise a two C2 domain-containing protein family and are known to aggregate phosphatidylserine membranes in a calcium-dependent manner. No enzymatic function has been attributed to copines yet. Due to a cross-reacting activity of Mikbeta1, an antibody to the IL-2Rbeta chain, we were able to serendipitously purify, partially microsequence, and clone human copine III. The 5 kb copine III transcript is expressed ubiquitously as determined by a multitissue Northern blot analysis. Phosphoamino acid analysis revealed phosphorylation of copine III on serine and threonine residues. In vitro kinase assays were performed with immunoprecipitated endogenous copine III, chromatography-purified endogenous copine III, and recombinant copine III expressed in Saccharomyces cerevisiae. The exogenous substrate myelin basic protein was phosphorylated in all in vitro kinase assays containing copine III immunoprecipitate or purified copine III. A 60-kDa band was observed in corresponding in gel kinase assays with staurosporine-activated cells. Cell lines expressing high levels of copine III protein had correspondingly high kinase activity in copine III antiserum immunoprecipitate. However, the copine amino acid sequences lack the traditional kinase catalytic domain. Therefore, the data suggest copine III may possess an intrinsic kinase activity and represent a novel unconventional kinase family. PMID- 11041870 TI - A high-affinity fluorenone-based beta 2-adrenergic receptor antagonist with a photoactivatable pharmacophore. AB - To develop molecules capable of directly probing the catechol binding region of the beta(2)-adrenergic receptor (beta(2)AR), novel benzophenone- and fluorenone based beta(2)AR antagonists were prepared as potential photoaffinity probes. While the benzophenone-containing ligands bound with relatively modest affinity, one of the fluorenone-based compounds, 4-(2-hydroxy-3-isopropylaminopropoxy)-7 amino-6-iodofluorenone+ ++ (iodoaminoflisopolol, IAmF), showed very high affinity for the beta(2)AR, inhibiting [(125)I]ICYP binding with an apparent K(i) of approximately 1 x 10(-)(9) M. In comparison to the benzophenone ligands, the fluorenone ligands have one additional carbon-carbon bond that creates a planar unsaturated ring system and leads to a large increase in receptor binding affinity. Unlike previous beta(2)AR photoaffinity ligands, an attractive and unique feature of the fluorenone derivative IAmF is that the large planar unsaturated ring (believed to correspond to the catechol end of other beta(2)AR ligands) serves as both the binding pharmacophore and the photoreaction center for this molecule. With this potential for directly probing the catechol binding region of the beta(2)AR, we synthesized and tested IAmF in carrier-free radioiodinated form ([(125)I]IAmF). When photoreduction was conducted at 350 nm for 20 min, [(125)I]IAmF was able to produce cross-linked products in both triethylamine and methanol, with a reactivity pattern similar to that found in benzophenone photochemistry. As a final test of suitability as a photoaffinity label, specific labeling of the beta(2)AR in membranes (protectable by 10 microM alprenolol) was demonstrated. [(125)I]IAmF represents a new class of beta(2)AR photoaffinity labels that can directly probe the catechol-analogous antagonist pharmacophore binding site in the beta(2)AR ligand binding pocket. PMID- 11041871 TI - Effect of avidin on channel kinetics of biotinylated gramicidin. AB - Membrane protein functioning basically depends on the supramolecular structure of the proteins which can be modulated by specific interactions with external ligands. The effect of a water-soluble protein bearing specific binding sites on the kinetics of ionic channels formed by gramicidin A (gA) in planar bilayer lipid membranes (BLM) has been studied using three independent approaches: (1) sensitized photoinactivation, (2) single-channel, and (3) autocorrelation measurements of current fluctuations. As shown previously [Rokitskaya, T. I., et al. (1996) Biochim. Biophys. Acta 1275, 221], the time course of the flash induced current decrease in most cases follows a single-exponential decay with an exponential factor (tau) that corresponds to the gA single-channel lifetime. Addition of avidin does not affect tau for gA channels, but causes a dramatic increase in tau for channels formed by gA5XB, a biotinylated analogue of gA. This effect is reversed by addition of an excess of biotin to the bathing solution. The average single-channel duration of gA5XB was about 3.6 s as revealed by single-channel recording of the BLM current. After prolonged incubation with avidin, a long-lasting open state of the gA5XB channel appeared which did not close for more than 10 min. The data on gA5XB photoinactivation kinetics and single-channel measurements were confirmed by analysis of the corresponding power spectra of the current fluctuations obtained in the control, in the presence of avidin, and after the addition of biotin. We infer that avidin produces a deceleration of gA5XB channel kinetics by motional restriction of gA5XB monomers and dimers upon the formation of avidin and gA5XB complexes, which would stabilize the channel state and thus increase the single-channel lifetime. PMID- 11041872 TI - The conserved C-terminus of the citrate (CitP) and malate (MleP) transporters of lactic acid bacteria is involved in substrate recognition. AB - The membrane potential-generating transporters CitP of Leuconostoc mesenteroides and MleP of Lactococcus lactis are homologous proteins with 48% identical residues that catalyze citrate-lactate and malate-lactate exchange, respectively. The two transporters are highly specific for substrates containing a 2 hydroxycarboxylate motif (HO-CR(2)-COO(-)) in which substitutions of the R groups are tolerated well. Differences in substrate specificity between MleP and CitP are based on subtle changes in the interaction of the protein with the R groups affecting both binding and translocation properties. The conserved, 46-residue long C-terminal region of the transporters containing the C-terminal putative transmembrane segment XI was investigated for its role in substrate recognition by constructing chimeric transporters. Replacement of the C-terminal region of MleP with that of CitP and vice versa did not alter the exchange kinetics with the substrates malate and citrate, indicating that the main interactions between the proteins and di- and tricarboxylate substrates were not altered. In contrast, the interaction of the proteins with the monocarboxylate substrates mandelate and 2-hydroxyisovalerate changed in a complementary manner. The affinity of CitP for the S-enantiomers of these substrates was at least 1 order of magnitude lower than observed for MleP. Introduction of the C-terminal residues of MleP in CitP resulted in a higher affinity and vice versa. Interchanging the C-termini had a more complicated effect on the R-enantiomers, affecting different kinetic parameters with different substrates, indicating multiple interactions of the R groups at this side of the binding pocket. It is suggested that the binding pocket is located between transmembrane segment XI and the other transmembrane segments of the transporters. PMID- 11041873 TI - Mechanism for activation of mouse mast cell tryptase: dependence on heparin and acidic pH for formation of active tetramers of mouse mast cell protease 6. AB - Tryptase, a serine protease with trypsin-like substrate cleavage properties, is one of the key effector molecules during allergic inflammation. It is stored in large quantities in the mast cell secretory granules in complex with heparin proteoglycan, and these complexes are released during mast cell degranulation. In the present paper, we have studied the mechanism for tryptase activation. Recombinant mouse tryptase, mouse mast cell protease 6 (mMCP-6), was produced in a mammalian expression system. The mMCP-6 fusion protein contained an N-terminal 6 x His tag followed by an enterokinase (EK) site replacing the native activation peptide (6xHis-EK-mMCP-6). In the absence of heparin, barely detectable enzyme activity was obtained after enterokinase cleavage of 6xHis-EK-mMCP-6 over a pH range of 5.5-7.5. However, when heparin was present, 6xHis-EK-mMCP-6 yielded active enzyme when enterokinase cleavage was performed at pH 5.5-6.0 but not at neutral pH. Affinity chromatography analysis showed that mMCP-6 bound strongly to heparin-Sepharose at pH 6.0 but not at neutral pH. After enterokinase cleavage of the sample at pH 6.0, mMCP-6 occurred in inactive monomeric form as shown by FPLC analysis on a Superdex 200 column. When heparin was added at pH 6.0, enzymatically active higher molecular weight complexes were formed, e.g., a dominant approximately 200 kDa complex that may correspond to tryptase tetramers. No formation of active tetramers was observed at neutral pH. When injected intraperitoneally, mMCP-6 together with heparin caused neutrophil influx, but no signs of inflammation were seen in the absence of heparin. The present paper thus indicates a crucial role for heparin in the formation of active mast cell tryptase. PMID- 11041874 TI - An indel within the C8 alpha subunit of human complement C8 mediates intracellular binding of C8 gamma and formation of C8 alpha-gamma. AB - Human C8 is one of five complement components (C5b, C6, C7, C8, and C9) that interact to form the cytolytic membrane attack complex, or MAC. It is an oligomeric protein composed of three subunits (C8alpha, C8beta, C8gamma) that are products of different genes. In C8 from serum, these are arranged as a disulfide linked C8alpha-gamma dimer that is noncovalently associated with C8beta. In this study, the site on C8alpha that mediates intracellular binding of C8gamma to form C8alpha-gamma was identified. From a comparative analysis of indels (insertions/deletions) in C8alpha and its structural homologues C8beta, C6, C7, and C9, it was determined that C8alpha contains a unique insertion (residues 159 175), which includes Cys(164) that forms the disulfide bond to C8gamma. Incorporation of this sequence into C8beta and coexpression of the resulting construct (iC8beta) with C8gamma produced iC8beta-gamma, an atypical disulfide linked dimer. In related experiments, C8gamma was shown to bind noncovalently to mutant forms of C8alpha and iC8beta in which Cys(164)-->Gly(164) substitutions were made. In addition, C8gamma bound specifically to an immobilized synthetic peptide containing the mutant indel sequence. Together, these results indicate (a) intracellular binding of C8gamma to C8alpha is mediated principally by residues contained within the C8alpha indel, (b) binding is not strictly dependent on Cys(164), and (c) C8gamma must contain a complementary binding site for the C8alpha indel. PMID- 11041875 TI - Equilibrium and kinetic analysis of folding of ketosteroid isomerase from Comamonas testosteroni. AB - Equilibrium and kinetic analyses have been carried out to elucidate the folding mechanism of homodimeric ketosteroid isomerase (KSI) from Comamonas testosteroni. The folding of KSI was reversible since the activity as well as the fluorescence and CD spectra was almost completely recovered after refolding. The equilibrium unfolding transitions monitored by fluorescence and CD measurements were almost coincident with each other, and the transition midpoint increased with increasing protein concentration. This suggests that the KSI folding follows a simple two state mechanism consisting of native dimer and unfolded monomer without any thermodynamically stable intermediates. Sedimentation equilibrium analysis and size-exclusion chromatography of KSI at different urea concentrations supported the two-state model without any evidence of folded monomeric intermediates. Consistent with the two-state model, (1)H-(15)N HSQC spectra obtained for KSI in the unfolding transition region could be reproduced by a simple addition of the spectra of the native and the unfolded KSI. The KSI refolding kinetics as monitored by fluorescence intensity could be described as a fast first-order process followed by a second-order and a subsequent slow first-order processes with rate constants of 60 s(-)(1), 5.4 x 10(4) M(-)(1).s(-)(1), and 0.017 s( )(1), respectively, at 0.62 M urea, suggesting that there may be a monomeric folding intermediate. After a burst phase that accounts for >83% of the total amplitude, the negative molar ellipticity at 225 nm increased slowly in a single phase at a rate comparable to that of the bimolecular intermediate step. The kinetics of activity recovery from the denatured state were markedly dependent upon the protein concentration, implying that the monomers are not fully active. Taken together, our results demonstrate that the dimerization induces KSI to fold into the complete structure and is crucial for maintaining the tertiary structure to perform efficient catalysis. PMID- 11041876 TI - Complex waxes. PMID- 11041877 TI - Separable whorl-specific expression and negative regulation by enhancer elements within the AGAMOUS second intron. AB - We analyzed the 4-kb intragenic control region of the AGAMOUS (AG) gene to gain insight into the mechanisms controlling its expression during early flower development. We identified three major expression patterns conferred by 19 AG::reporter gene constructs: the normal AG pattern, a stamen-specific pattern, and a predominantly carpel pattern. To determine whether these three expression patterns were under negative control by APETALA2 (AP2) or LEUNIG (LUG), we analyzed beta-glucuronidase staining patterns in Arabidopsis plants homozygous for strong ap2 and lug mutations. Our results indicated that the stamen-specific pattern was independent of AP2 but dependent on LUG; conversely, the carpel specific pattern was independent of LUG but dependent on AP2. These results lead to a model of control of AG expression such that expression in each of the two inner whorls is under independent positive and negative control. PMID- 11041878 TI - Simulation of fungal-mediated cell death by fumonisin B1 and selection of fumonisin B1-resistant (fbr) Arabidopsis mutants. AB - Fumonisin B1 (FB1), a programmed cell death-eliciting toxin produced by the necrotrophic fungal plant pathogen Fusarium moniliforme, was used to simulate pathogen infection in Arabidopsis. Plants infiltrated with 10 microM FB1 and seedlings transferred to agar media containing 1 microM FB1 develop lesions reminiscent of the hypersensitive response, including generation of reactive oxygen intermediates, deposition of phenolic compounds and callose, accumulation of phytoalexin, and expression of pathogenesis-related (PR) genes. Arabidopsis FB1-resistant (fbr) mutants were selected directly by sowing seeds on agar containing 1 microM FB1, on which wild-type seedlings fail to develop. Two mutants chosen for further analyses, fbr1 and fbr2, had altered PR gene expression in response to FB1. fbr1 and fbr2 do not exhibit differential resistance to the avirulent bacterial pathogen Pseudomonas syringae pv maculicola (ES4326) expressing the avirulence gene avrRpt2 but do display enhanced resistance to a virulent isogenic strain that lacks the avirulence gene. Our results demonstrate the utility of FB1 for high-throughput isolation of Arabidopsis defense-related mutants and suggest that pathogen-elicited programmed cell death of host cells may be an important feature of compatible plant-pathogen interactions. PMID- 11041880 TI - A leaf-derived signal is a quantitative determinant of floral form in Impatiens. AB - The completion of flower development in Impatiens balsamina requires continuous inductive (short-day) conditions. We have previously shown that a leaf-derived signal has a role in floral maintenance. The research described here analyzes the role of the leaf in flower development. Leaf removal treatments, in which plants were restricted to a specified number of leaves, resulted in flowers with increased petal number, up to double that of the undefoliated control. Similar petal number increases (as well as changes in bract number or morphology) were recorded when plants began their inductive treatment at a late developmental age or when plants of a nonreverting line (capable of floral maintenance in the absence of continuous short days) were transferred from short days to long days. Our data imply that the increased petal number was neither a response to stress effects associated with leaf removal nor a result of alterations in primordium initiation rates or substitutions of petals for stamens. Rather, the petal initiation phase was prolonged when the amounts of a leaf-derived signal were limiting. We conclude that a leaf-derived signal has a continuous and quantitative role in flower development and propose a temporal model for the action of organ identity genes in Impatiens. This work adds a new dimension to the prevailing ABC model of flower development and may provide an explanation for the wide variety and instabilities of floral form seen among certain species in nature. PMID- 11041879 TI - Fumonisin B1-induced cell death in arabidopsis protoplasts requires jasmonate-, ethylene-, and salicylate-dependent signaling pathways. AB - We have established an Arabidopsis protoplast model system to study plant cell death signaling. The fungal toxin fumonisin B1 (FB1) induces apoptosis-like programmed cell death (PCD) in wild-type protoplasts. FB1, however, only marginally affects the viability of protoplasts isolated from transgenic NahG plants, in which salicylic acid (SA) is metabolically degraded; from pad4-1 mutant plants, in which an SA amplification mechanism is thought to be impaired; or from jar1-1 or etr1-1 mutant plants, which are insensitive to jasmonate (JA) or ethylene (ET), respectively. FB1 susceptibility of wild-type protoplasts decreases in the dark, as does the cellular content of phenylalanine ammonia lyase, a light-inducible enzyme involved in SA biosynthesis. Interestingly, however, FB1-induced PCD does not require the SA signal transmitter NPR1, given that npr1-1 protoplasts display wild-type FB1 susceptibility. Arabidopsis cpr1-1, cpr6-1, and acd2-2 protoplasts, in which the SA signaling pathway is constitutively activated, exhibit increased susceptibility to FB1. The cpr6-1 and acd2-2 mutants also constitutively express the JA and ET signaling pathways, but only the acd2-2 protoplasts undergo PCD in the absence of FB1. These results demonstrate that FB1 killing of Arabidopsis is light dependent and requires SA-, JA-, and ET-mediated signaling pathways as well as one or more unidentified factors activated by FB1 and the acd2-2 mutation. PMID- 11041881 TI - Ozone-sensitive arabidopsis rcd1 mutant reveals opposite roles for ethylene and jasmonate signaling pathways in regulating superoxide-dependent cell death. AB - We have isolated a codominant Arabidopsis mutant, radical-induced cell death1 (rcd1), in which ozone (O(3)) and extracellular superoxide (O(2)(*)-), but not hydrogen peroxide, induce cellular O(2)(*)- accumulation and transient spreading lesions. The cellular O(2)(*)- accumulation is ethylene dependent, occurs ahead of the expanding lesions before visible symptoms appear, and is required for lesion propagation. Exogenous ethylene increased O(2)(*)--dependent cell death, whereas impairment of ethylene perception by norbornadiene in rcd1 or ethylene insensitivity in the ethylene-insensitive mutant ein2 and in the rcd1 ein2 double mutant blocked O(2)(*)- accumulation and lesion propagation. Exogenous methyl jasmonate inhibited propagation of cell death in rcd1. Accordingly, the O(3) exposed jasmonate-insensitive mutant jar1 displayed spreading cell death and a prolonged O(2)(*)- accumulation pattern. These results suggest that ethylene acts as a promoting factor during the propagation phase of developing oxyradical dependent lesions, whereas jasmonates have a role in lesion containment. Interaction and balance between these pathways may serve to fine-tune propagation and containment processes, resulting in alternate lesion size and formation kinetics. PMID- 11041882 TI - The TT8 gene encodes a basic helix-loop-helix domain protein required for expression of DFR and BAN genes in Arabidopsis siliques. AB - The TRANSPARENT TESTA8 (TT8) locus is involved in the regulation of flavonoid biosynthesis in Arabidopsis. The tt8-3 allele was isolated from a T-DNA mutagenized Arabidopsis collection and found to be tagged by an integrative molecule, thus permitting the cloning and sequencing of the TT8 gene. TT8 identity was confirmed by complementation of tt8-3 and sequence analysis of an additional allele. The TT8 gene encodes a protein that displays a basic helix loop-helix at its C terminus and represents an Arabidopsis ortholog of the maize R transcription factors. The TT8 transcript is present in developing siliques and in young seedlings. The TT8 protein is required for normal expression of two flavonoid late biosynthetic genes, namely, DIHYDROFLAVONOL 4-REDUCTASE (DFR) and BANYULS (BAN), in Arabidopsis siliques. Interestingly, TRANSPARENT TESTA GLABRA1 (TTG1) and TT2 genes also control the expression of DFR and BAN genes. Our results suggest that the TT8, TTG1, and TT2 proteins may interact to control flavonoid metabolism in the Arabidopsis seed coat. PMID- 11041883 TI - Regulation of gynoecium marginal tissue formation by LEUNIG and AINTEGUMENTA. AB - The carpel is the female reproductive organ of flowering plants. In Arabidopsis, congenital fusion of two carpels leads to the formation of an enclosed gynoecium. The margins of the two fused carpels are meristematic in nature and give rise to placentas, ovules, septa, abaxial repla, and the majority of the stylar and stigmatic tissues. Thus, understanding how the marginal tissues are specified and identifying genes that direct their development may provide important insight into higher plant reproductive development. In this study, we show that LEUNIG and AINTEGUMENTA are two critical regulators of marginal tissue development. Double mutants of leunig aintegumenta fail to develop placentas, ovules, septa, stigma, and style. This effect is specific to the leunig aintegumenta double mutant and is not found in other double mutant combinations such as leunig apetala2 or aintegumenta apetala2. Additional analyses indicate that the absence of marginal tissues in leunig aintegumenta double mutants is not mediated by ectopic AGAMOUS. We propose that LEUNIG and AINTEGUMENTA act together to control the expression of common target genes that regulate cell proliferation associated with marginal tissue development. PMID- 11041884 TI - GRCD1, an AGL2-like MADS box gene, participates in the C function during stamen development in Gerbera hybrida. AB - Despite the differences in flower form, the underlying mechanism in determining the identity of floral organs is largely conserved among different angiosperms, but the details of how the functions of A, B, and C are specified varies greatly among plant species. Here, we report functional analysis of a Gerbera MADS box gene, GRCD1, which is orthologous to AGL2-like MADS box genes. Members of this group of genes are being reported in various species in growing numbers, but their functions remained largely unsettled. GRCD1 expression is detected in all four whorls, but the strongest signal is seen in the developing stamen and carpel. Downregulating GRCD1 expression by antisense transformation revealed that lack of GRCD1 caused homeotic changes in one whorl only: sterile staminodes, which normally develop in whorl 3 of marginal female florets, were changed into petals. This indicates that the GRCD1 gene product is active in determining stamen identity. Transgenic downregulation of GRCD1 causes a homeotic change similar to that in the downregulation of the Gerbera C function genes GAGA1 and GAGA2, but one that is limited to whorl 3. Downregulation of GRCD1 expression does not reduce expression of GAGA1 or GAGA2, or vice versa; and in yeast two hybrid analysis, GRCD1 is able to interact with GAGA1 and GAGA2. We propose that a heterodimer between the GRCD1 and GAGA1/2 gene products is needed to fulfill the C function in whorl 3 in Gerbera. PMID- 11041886 TI - HRT gene function requires interaction between a NAC protein and viral capsid protein to confer resistance to turnip crinkle virus. AB - An Arabidopsis protein was found to interact specifically with the capsid protein (CP) of turnip crinkle virus (TCV) through yeast two-hybrid screening. This protein, designated TIP (for TCV-interacting protein), was found to be a member of the recently recognized NAC family of proteins. NAC proteins have been implicated in the regulation of development of plant embryos and flowers. TIP alone was able to activate expression of reporter genes in yeast if fused to a DNA binding domain, suggesting that it may be a transcriptional activator. The TIP binding region in the TCV CP has been mapped to the N-terminal 25 amino acids. Site-directed mutagenesis within this region revealed that loss of the TIP CP interaction in the yeast two-hybrid assay correlated with loss of the ability of TCV to induce the hypersensitive response and resistance in the TCV-resistant Arabidopsis ecotype Dijon (Di-0 and its inbred line Di-17). These data suggest that TIP is an essential component in the TCV resistance response pathway. PMID- 11041885 TI - AHM1, a novel type of nuclear matrix-localized, MAR binding protein with a single AT hook and a J domain-homologous region. AB - Interactions between the nuclear matrix and special regions of chromosomal DNA called matrix attachment regions (MARs) have been implicated in various nuclear functions. We have identified a novel protein from wheat, AT hook-containing MAR binding protein1 (AHM1), that binds preferentially to MARs. A multidomain protein, AHM1 has the special combination of a J domain-homologous region and a Zn finger-like motif (a J-Z array) and an AT hook. For MAR binding, the AT hook at the C terminus was essential, and an internal portion containing the Zn finger like motif was additionally required in vivo. AHM1 was found in the nuclear matrix fraction and was localized in the nucleoplasm. AHM1 fused to green fluorescent protein had a speckled distribution pattern inside the nucleus. AHM1 is most likely a nuclear matrix component that functions between intranuclear framework and MARs. J-Z arrays can be found in a group of (hypothetical) proteins in plants, which may share some functions, presumably to recruit specific Hsp70 partners as co-chaperones. PMID- 11041887 TI - The pea light-independent photomorphogenesis1 mutant results from partial duplication of COP1 generating an internal promoter and producing two distinct transcripts. AB - The pea lip1 (light-independent photomorphogenesis1) mutant shows many of the characteristics of light-grown development when grown in continuous darkness. To investigate the identity of LIP1, cDNAs encoding the pea homolog of COP1, a repressor of photomorphogenesis identified in Arabidopsis, were isolated from wild-type and lip1 pea seedlings. lip1 seedlings contained a wild-type COP1 transcript as well as a larger COP1' transcript that contained an internal in frame duplication of 894 bp. The COP1' transcript segregated with the lip1 phenotype in F(2) seedlings and could be translated in vitro to produce a protein of approximately 100 kD. The COP1 gene in lip1 peas contained a 7.5-kb duplication, consisting of exons 1 to 7 of the wild-type sequence, located 2.5 kb upstream of a region of genomic DNA identical to the wild-type COP1 DNA sequence. Transcription and splicing of the mutant COP1 gene was predicted to produce the COP1' transcript, whereas transcription from an internal promoter in the 2.5-kb region of DNA located between the duplicated regions of COP1 would produce the wild-type COP1 transcript. The presence of small quantities of wild-type COP1 transcripts may reduce the severity of the phenotype produced by the mutated COP1' protein. The genomic DNA sequences of the COP1 gene from wild-type and lip1 peas and the cDNA sequences of COP1 and COP1' transcripts have been submitted to the EMBL database under the EMBL accession numbers AJ276591, AJ276592, AJ289773, and AJ289774, respectively. PMID- 11041888 TI - Glutathione and a UV light-induced glutathione S-transferase are involved in signaling to chalcone synthase in cell cultures. AB - UV irradiation stimulates expression of the gene encoding the key enzyme chalcone synthase (CHS), which leads to the generation of protective flavonoids in parsley cell cultures. CHS transcripts increase after 3 to 4 hr, and early genes are involved in the signal transduction to the CHS promoter. By using the fluorescent differential display technique in a large-scale screening, several early UV light induced genes were isolated. Of these, a novel glutathione S-transferase (PcGST1) is induced within 2 hr and precedes CHS expression. Overexpression of PcGST1 in transformed cell lines containing a CHS promoter/luciferase reporter (CHS-LUC) affected the onset of LUC transcription. Supplementing these cell lines with glutathione immediately stimulated CHS-LUC expression within 2 hr in dark incubated cells and resulted in a biphasic induction profile in UV-irradiated cells. Our data indicate the involvement of glutathione and PcGST1 in early events of a UV light-dependent signal transduction pathway to CHS. In this context, the oxidative status of a cell acts as a central regulating element. PMID- 11041889 TI - Insertion of OEP14 into the outer envelope membrane is mediated by proteinaceous components of chloroplasts. AB - Most chloroplastic outer envelope membrane proteins are synthesized in the cytosol at their mature size without a cleavable targeting signal. Their insertion into the outer membrane is insensitive to thermolysin pretreatment of chloroplasts and does not require ATP. The insertion has been assumed to be mediated by a spontaneous mechanism or by interaction solely with the lipid components of the outer membrane. However, we show here that insertion of an outer membrane protein requires some trypsin-sensitive and some N-ethylmaleimide sensitive components of chloroplasts. Association and insertion of the outer membrane protein are saturable and compete with the import of another outer membrane protein. These data suggest that import of chloroplastic outer membrane proteins occurs at specific proteinaceous sites on chloroplasts. PMID- 11041890 TI - Genetic interactions during root hair morphogenesis in Arabidopsis. AB - Root hairs are a major site for the uptake of water and nutrients into plants and form an increasingly important model system for studies of development of higher plants and cell biology. We have identified loss-of-function mutations in eight new genes required for hair growth in Arabidopsis: SHAVEN1 (SHV1), SHV2, and SHV3; CENTIPEDE1 (CEN1), CEN2, and CEN3; BRISTLED1 (BST1); and SUPERCENTIPEDE1 (SCN1). We combined mutations in 79 pairs of genes to determine the stages at which these and six previously known genes contribute to root hair formation. Double mutant phenotypes revealed roles for several genes that could not have been predicted from the single mutant phenotypes. For example, we show that TIP1 and RHD3 are required much earlier in hair formation than previous studies have suggested. We present a genetic model for root hair morphogenesis that defines the roles of each gene, and we suggest hypotheses about functional relationships between genes. PMID- 11041891 TI - Complex spatial responses to cucumber mosaic virus infection in susceptible Cucurbita pepo cotyledons. AB - Cucumber mosaic virus infection of its susceptible host Cucurbita pepo results in a program of biochemical changes after virus infection. Applying a spatial analysis to expanding infected lesions, we investigated the relationship between the changes in enzyme activity and gene expression. Patterns of altered expression were seen that could not be detected by RNA gel blot analysis. For all the host genes studied, there was a downregulation (shutoff) of expression within the lesion. In addition, two distinct types of upregulation were observed. The expression of heat shock protein 70 (HSP70) and NADP(+)-dependent malic enzyme (NADP-ME) showed induction in apparently uninfected cells ahead of the infection. This response was more localized than the upregulation exhibited by catalase expression, which occurred throughout the uninfected regions of the tissue. The experiments showed that virus infection induced immediate and subsequent changes in gene expression by the host and that the infection has the potential to give advance signaling of the imminent infection. PMID- 11041892 TI - Cell cycle regulation of the tobacco ribonucleotide reductase small subunit gene is mediated by E2F-like elements. AB - Ribonucleotide reductase (RNR) is a key enzyme involved in the DNA synthesis pathway. The RNR-encoded genes are cell cycle regulated and specifically expressed in S phase. The promoter of the RNR2 gene encoding for the small subunit was isolated from tobacco. Both in vivo and in vitro studies of the DNA protein interactions in synchronized BY2 tobacco cells showed that two E2F-like motifs were involved in multiple specific complexes, some of which displayed cell cycle-regulated binding activities. Moreover, these two elements could specifically interact with a purified tobacco E2F protein. Involvement of the E2F elements in regulating the RNR2 promoter was checked by functional analyses in synchronized transgenic BY2 cells transformed with various RNR2 promoter constructs fused to the luciferase reporter gene. The two E2F elements were involved in upregulation of the promoter at the G1/S transition and mutation of both elements prevented any significant induction of the RNR promoter. In addition, one of the E2F elements sharing homology with the animal E2F/cell cycle dependent element motif behaved like a repressor when outside of the S phase. These data provide evidence that E2F elements play a crucial role in cell cycle regulation of gene transcription in plants. PMID- 11041893 TI - Alterations in CER6, a gene identical to CUT1, differentially affect long-chain lipid content on the surface of pollen and stems. AB - Very long chain lipids contribute to the hydrophobic cuticle on the surface of all land plants and are an essential component of the extracellular pollen coat in the Brassicaceae. Mutations in Arabidopsis CER genes eliminate very long chain lipids from the cuticle surface and, in some cases, from the pollen coat. In Arabidopsis, the loss of pollen coat lipids can disrupt interactions with the stigma, inhibiting pollen hydration and causing sterility. We have positionally cloned CER6 and demonstrate that a wild-type copy complements the cer6-2 defect. In addition, we have identified a fertile, intragenic suppressor, cer6-2R, that partially restores pollen coat lipids but does not rescue the stem wax defect, suggesting an intriguing difference in the requirements for CER6 activity on stems and the pollen coat. Importantly, analysis of this suppressor demonstrates that low amounts of very long chain lipids are sufficient for pollen hydration and germination. The predicted CER6 amino acid sequence resembles that of fatty acid-condensing enzymes, consistent with its role in the production of epicuticular and pollen coat lipids >28 carbons long. DNA sequence analysis revealed the nature of the cer6-1, cer6-2, and cer6-2R mutations, and segregation analysis showed that CER6 is identical to CUT1, a cDNA previously mapped to a different chromosome arm. Instead, we have determined that a new gene, CER60, with a high degree of nucleotide and amino acid similarity to CER6, resides at the original CUT1 locus. PMID- 11041894 TI - A Jewish physician amidst the Holocaust. AB - On November 27, 1940, in Nazi Germany, a prescription was written by a Jewish physician, Dr Lucie Adelsberger. This article examines that prescription and tells the story of the physician who wrote it. Tracing the fate of Dr Adelsberger throughout the Holocaust, this article describes her ordeal as a prisoner physician at the Auschwitz concentration camp and her struggles to help sick camp inmates survive under the most brutal of conditions. Arch Intern Med. 2000;160:2891-2894 PMID- 11041895 TI - The heart and estrogen/progestin replacement study revisited: hormone replacement therapy produced net harm, consistent with the observational data. AB - Lower coronary event rates in women receiving hormone replacement therapy (HRT) have led to a presumption of benefit. The Heart and Estrogen/Progestin Replacement Study, a large randomized trial, observed a 1.4% first year excess of coronary events, well beyond the plausible play of chance on the expected effect. Over the duration of the study, event totals were similar, but patients treated with HRT experienced them earlier, with a net loss of patient-months of event free survival. The point at which the lower event rate in hormone-treated patients would fully repay the first year loss, with constant rates, is almost double the trial duration (of 4.1 years). Since patients in the trial were preselected for satisfactory adherence to therapy, the net benefit in practice is likely to be even less. Had the patients in the Heart and Estrogen/Progestin Replacement Study been recruited to an observational study at various intervals over the first 5 years after starting HRT, the apparent risk reduction over 5 years would have been between 21% and 34%. A previous meta-analysis of trials of HRT for other indications also shows net harm. Women with or at high risk of coronary heart disease should not start HRT. There is a risk that women without coronary heart disease might experience even greater net harm from HRT. The late benefit is necessarily limited, as it cannot exceed the event rate. The mechanism of the early loss is unknown; if it were reduced proportionately less than the late benefit, considerable net harm could result. Arch Intern Med. 2000;160:2897 2900 PMID- 11041896 TI - Friend or foe? How primary care physicians perceive hospitalists. AB - BACKGROUND: Increased use of hospitalists is redefining the role of primary care physicians. Whether primary care physicians welcome this transition is unknown. We examined primary care physicians' perceptions of how hospitalists affect their practices, their patient relationships, and overall patient care. METHODS: A mailed survey of randomly selected general internists, general pediatricians, and family practitioners with experience with hospitalists practicing in California. MAIN OUTCOME MEASURES: Physicians' self-reports of hospitalists' effects on quality of patient care and on their own practices. RESULTS: Seven hundred eight physicians were eligible for this study, and there was a 74% response rate. Of the 524 physicians who responded, 34% were internists, 38% were family practitioners, and 29% were pediatricians. Of the 524 respondents, 335 (64%) had hospitalists available to them and 120 (23%) were required to use hospitalists for all admissions. Physicians perceived hospitalists as increasing (41%) or not changing (44%) the overall quality of care and perceived their practice style differences as neutral or beneficial. Twenty-eight percent of primary care physicians believed that the quality of the physician-patient relationship decreased; 69% reported that hospitalists did not affect their income; 53% believed that hospitalists decreased their workload; and 50% believed that hospitalists increased practice satisfaction. In a multivariate model predicting physician perceptions, internists, physicians who attributed loss of income to hospitalists, and physicians in mandatory hospitalist systems viewed hospitalists less favorably. CONCLUSIONS: Practicing primary care physicians have generally favorable perceptions of hospitalists' effect on patients and on their own practice satisfaction, especially in voluntary hospitalist systems that decrease the workload of primary care physicians and do not threaten their income. Primary care physicians, particularly internists, are less accepting of mandatory hospitalist systems. Arch Intern Med. 2000;160:2902-2908 PMID- 11041897 TI - Physicians' attitudes about involvement in lethal injection for capital punishment. AB - BACKGROUND: Physicians could play various roles in carrying out capital punishment via lethal injection. Medical societies like the American Medical Association (AMA) and American College of Physicians have established which roles are acceptable and which are disallowed. No one has explored physicians' attitudes toward their potential roles in this process. METHODS: We surveyed physicians about how acceptable it was for physicians to engage in 8 actions disallowed by the AMA and 4 allowed actions involving lethal injection. Questions assessing attitudes toward capital punishment and assisted suicide were included. The impact of attitudinal and demographic variables on the number of disallowed actions deemed acceptable was analyzed via analysis of variance and multiple logistic regression analysis. RESULTS: Four hundred eighty-two physicians (51%) returned questionnaires. Eighty percent indicated that at least 1 of the disallowed actions was acceptable, 53% indicated that 5 or more were acceptable, and 34% approved all 8 disallowed actions. The percentage of respondents approving of disallowed actions varied from 43% for injecting lethal drugs to 74% for determining when death occurred. All 4 allowed actions were deemed acceptable by the majority of respondents. Favoring the death penalty (P<.001) and the acceptance of assisted suicide (P<.001) were associated with an increased number of disallowed actions that were deemed acceptable. CONCLUSIONS: Despite medical society policies, the majority of physicians surveyed approved of most disallowed actions involving capital punishment, indicating that they believed it is acceptable in some circumstances for physicians to kill individuals against their wishes. It is possible that the lack of stigmatization by colleagues allows physicians to engage in such practices. Arch Intern Med. 2000;160:2912-2916 PMID- 11041898 TI - Vertebral fracture risk with long-term corticosteroid therapy: prevalence and relation to age, bone density, and corticosteroid use. AB - BACKGROUND: Few data are available regarding vertebral fracture risk in patients treated with oral corticosteroids. The aim of this study was to determine the prevalence and the role of risk factors such as age, bone mineral density (BMD), and corticosteroid use for vertebral deformity in patients receiving long-term corticosteroid therapy. METHODS: Thoracolumbar x-ray films, BMD, and details on corticosteroid use were obtained on 229 consecutive patients treated with long term corticosteroid regimens (> or = 6 months of prednisone, > or = 5mg/d or equivalent) seen at 4 referral centers. Comparisons were made with a population control group of 286 male and female controls not taking corticosteroids (aged > or = 60 years). RESULTS: Sixty-five patients (28%) had at least 1 vertebral deformity and 25 (11%) had 2 or more vertebral deformities. Older age, independent of BMD, was a significant risk factor for deformity. Patients aged 70 to 79 years had a 5-fold increased risk of deformity compared with patients younger than 60 years (odds ratio, 5.13; 95% confidence interval, 2.03-13.0). Compared with the population controls, the prevalence of deformities increased to a greater extent with each decade of age in the corticosteroid group (P =.005). Mean lumbar spine and femoral neck BMD Z scores were lower in the steroid-treated patients with deformities compared with the nonsteroid control group with deformities. When the effects of age, sex, body mass index, and duration of corticosteroid use were adjusted for in logistic regression analysis, low BMD was a modest predictor of deformity (for a 1-SD decrease in lumbar spine BMD: odds ratio, 1.31; 95% confidence interval, 1.02-1.68) and for a 1-SD decrease in femoral neck BMD: odds ratio, 1.77; 95% confidence interval, 1.07-2.94). CONCLUSIONS: The combination of increasing age and corticosteroid use is associated with a marked increase in the risk of vertebral deformity. Elderly patients commencing long-term corticosteroid therapy should be considered for antiosteoporotic therapy independently of their BMD. Arch Intern Med. 2000;160:2917-2922 PMID- 11041899 TI - Effectiveness and economic impact associated with a program for outpatient management of acute deep vein thrombosis in a group model health maintenance organization. AB - BACKGROUND: Controlled clinical trials have demonstrated that outpatient administration of low-molecular-weight heparin to patients with acute deep vein thrombosis (DVT) provides safety and efficacy equivalent to that of traditional inpatient therapy with unfractionated heparin. Whether favorable results reported in controlled clinical trials are achievable in clinical practice is an important consideration. METHODS: Appropriate patients with objectively diagnosed DVT were treated as outpatients with low-molecular-weight heparin and warfarin sodium according to an approved guideline. The primary end point for analysis consisted of objectively diagnosed symptomatic recurrent thromboembolism or major bleeding within a 90-day evaluation period. The incremental cost incurred by the organization while using the outpatient DVT treatment guideline was determined. Incremental cost savings of the outpatient DVT treatment program were determined based on the cost that would have accrued had the patient been admitted to the hospital for treatment with unfractionated heparin. RESULTS: We enrolled 391 patients (91.4%) in the outpatient DVT treatment program. Of these, 373 (95.4%) completed 90 days of therapy without reaching the primary end point. The percentage of patients reaching the primary outcome measure (4.6%) fell within the range of patients enrolled in controlled clinical trials (3.5%-9.4%). During the 2-year program evaluation, total cost savings of $1,108,587 were realized. CONCLUSIONS: Outpatient treatment of acute DVT can be managed safely and effectively in clinical practice. The potential savings associated with outpatient DVT treatment are substantial. Arch Intern Med. 2000;160:2926-2932 PMID- 11041900 TI - Determinants of peripheral arterial disease in the elderly: the Rotterdam study. AB - OBJECTIVE: To examine which atherosclerotic risk factors are determinants for peripheral arterial disease (PAD), we performed a population-based study in 6450 subjects (40% men, 60% women) aged 55 years and older. METHODS: The presence of PAD was assessed by measuring the ankle-arm systolic blood pressure index (AAI); PAD was considered present if the AAI was lower than 0.90 in either leg. In addition, a threshold AAI of 0.70 in either leg defined severe PAD. RESULTS: Determinants strongly and independently associated with PAD were age of at least 75 years (odds ratio [OR], 1.2; 95% confidence interval [CI], 1.0-1.6), fibrinogen level (OR, 1.5; 95% CI, 1.3-1.7), cigarette smoking (OR, 2.8; 95% CI, 2.3-3.4), diabetes mellitus (OR, 2.0; 95% CI, 1.6-2.5), and systolic blood pressure (OR, 1.2; 95% CI, 1.1-1.2). An inverse relation of high-density lipoprotein cholesterol level with PAD (OR, 0.7; 95% CI, 0.5-0.8) was found. Similar results were demonstrated for severe PAD. Separate analyses for men and women did not demonstrate differences in risk factors for PAD. CONCLUSIONS: Assessment of a wide range of atherosclerotic risk factors enabled us to quantify the relative importance of each factor as determinant for PAD. In total, 69% of the occurrence of PAD is attributable to cardiovascular risk factors measured in our study; smoking accounted for most (etiologic fraction, 18.1%). The results suggest that preventive management of PAD should be directed at systolic blood pressure, fibrinogen level, smoking, high-density lipoprotein cholesterol level, and diabetes mellitus. Arch Intern Med. 2000;160:2934-2938 PMID- 11041901 TI - New transient ischemic attack and stroke: outpatient management by primary care physicians. AB - BACKGROUND: Patients with transient ischemic attack (TIA) or stroke frequently first contact their primary care physician rather than seeking care at a hospital emergency department. The purpose of the present study was to identify a group of patients seen by primary care physicians in an office setting for a first-ever TIA or stroke and characterize their evaluation and management. METHODS: Practice audit based on retrospective, structured medical record abstraction from 27 primary care medical practices in 2 geographically separate communities in the eastern United States. RESULTS: Ninety-five patients with a first-ever TIA and 81 with stroke were identified. Seventy-nine percent of those with TIA vs 88% with stroke were evaluated on the day their symptoms occurred (P =.12). Only 6% were admitted to a hospital for evaluation and treatment on the day of the index visit (2% TIA; 10% stroke; P =.03); only an additional 3% were admitted during the subsequent 30 days. Specialists were consulted for 45% of patients. A brain imaging study (computed tomography or magnetic resonance imaging) was ordered on the day of the index visit in 30% (23% TIA, 37% stroke; P =.04), regardless of whether the patient was referred to a specialist. Carotid ultrasound studies were obtained in 28% (40% TIA, 14% stroke; P<.001), electrocardiograms in 19% (18% TIA, 21% stroke; P =.60), and echocardiograms in 16% (19% TIA, 14% stroke; P =.34). Fewer than half of patients with a prior history of atrial fibrillation (n = 24) underwent anticoagulation when evaluated at the index visit. Thirty-two percent of patients (31% TIA, 33% stroke; P =.70) were not hospitalized and had no evaluations performed during the first month after presenting to a primary care physician with a first TIA or stroke. Of these patients, 59% had a change in antiplatelet therapy on the day of the index visit. CONCLUSIONS: Further primary care physician education regarding the importance of promptly and fully evaluating patients with TIA or stroke may be warranted, and barriers to implementation of established secondary stroke prevention strategies need to be carefully explored. Arch Intern Med. 2000;160:2941-2946 PMID- 11041902 TI - Safety and efficacy of meloxicam in the treatment of osteoarthritis: a 12-week, double-blind, multiple-dose, placebo-controlled trial. The Meloxicam Osteoarthritis Investigators. AB - BACKGROUND: Meloxicam (Mobic; Boehringer Ingelheim, Ridgefield, Conn) is an enolic acid derivative of the oxicam group of nonsteroidal anti-inflammatory drugs (NSAIDs) whose mechanism of action may be related to prostaglandin (cyclooxygenase) synthetase inhibition. In previous studies, meloxicam has been found to be safe and effective in the treatment of osteoarthritis (OA) at doses of 7.5 to 15 mg daily. To evaluate a lower dose and a different patient population, we evaluated the efficacy and safety of 3 doses of meloxicam vs placebo and diclofenac for the treatment of OA among patients with symptom exacerbations. METHODS: In this double-blind, double-dummy, parallel-group, multicenter study, 774 patients with confirmed OA of the hip or knee and a flare were randomized and treated with daily oral administration of meloxicam tablets (at dosages of 3.75, 7.5, or 15 mg/d), diclofenac (100 mg [50 mg twice daily]), or placebo. Treatment was for 12 weeks, with regular assessments for drug safety and efficacy. Safety was assessed by evaluation of adverse events, vital signs, and laboratory data. Primary efficacy variables included the Western Ontario and McMaster University Osteoarthritis (WOMAC) index, the patient's overall assessment of pain, and the patient's and investigator's overall assessment of disease activity. RESULTS: The incidence of all adverse events was lower at each dosage of meloxicam than for diclofenac but greater than for placebo. However, the incidence of gastrointestinal adverse events and dropout rates because of such events was the same for meloxicam as for placebo and lower than for diclofenac. Meloxicam, at 7.5 and 15 mg/d, and diclofenac were statistically significantly more effective than placebo for all end points, while the 3.75-mg/d dosage of meloxicam did not always reach statistical significance for all end points. Efficacy was evident after 2 weeks of treatment, improved with increasing doses, and was maintained until the end of the trial. CONCLUSIONS: Meloxicam is a safe and effective medication for the symptomatic treatment of OA. The data support consideration of 7.5 to 15 mg of meloxicam once daily to treat the pain and stiffness of OA, with gastrointestinal tolerability comparable to that of placebo. Arch Intern Med. 2000;160:2947-2954 PMID- 11041903 TI - Diagnostic efficacy of unconjugated plasma metanephrines for the detection of pheochromocytoma. AB - BACKGROUND: Recently, measurement of plasma metanephrines was suggested to improve the detection of pheochromocytoma compared with the other common biochemical tests. OBJECTIVE: To examine the diagnostic precision of measurements of plasma metanephrines, plasma catecholamines, and urinary catecholamines and to assess their variability. METHODS: Plasma metanephrine as well as plasma and urinary catecholamine concentrations were measured by high-performance liquid chromatography with electrochemical detection. Before surgery, responses of plasma metanephrines and catecholamines to change of posture were determined. Intraoperatively, metanephrines and catecholamines were measured before skin incision, during maximal mechanical tumor manipulation, and repetitively after the tumor was separated from the circulation. Patients were reexamined 1 and 3 months after surgery. Patients with pheochromocytoma (n = 17) and with histologically proved other adrenal tumors (n = 14) were studied before, during, and after surgery. RESULTS: Measurement of plasma metanephrines and plasma and urinary catecholamines provided 100% and 82% sensitivity, respectively, for the detection of pheochromocytoma (P<.001). Levels of plasma catecholamines but not metanephrines increased in response to change of posture (norepinephrine, P =.03; epinephrine, P =.07) and intraoperative stress (norepinephrine, P =.002; epinephrine, P =.009). CONCLUSIONS: Plasma metanephrines offer improved efficacy for the diagnosis of pheochromocytoma. Less variability in response to external factors may favor plasma metanephrines in the screening for this disease. Arch Intern Med. 2000;160:2957-2963 PMID- 11041904 TI - The detection of dementia in the primary care setting. AB - BACKGROUND: Recognition and medical record documentation of dementia in the primary care setting are thought to be poor. To our knowledge, previous studies have not examined these issues in private practice office settings within the United States. OBJECTIVE: To determine the rate of unrecognized and undocumented dementia in a primary care internal medicine private practice. METHODS: This was a cross-sectional study of 297 ambulatory persons aged 65 years and older attending an internal medicine private group practice within an Asian American community of Honolulu, Hawaii. Of the subjects, 95% had been with their current primary care physician for at least 1 year. Each subject's primary care physician noted the presence or absence of dementia by questionnaire at the time of an office visit. An investigating physician (V.G.V.) subsequently assessed cognitive function using the Cognitive Abilities Screening Instrument, and confirmed the presence of dementia and its severity, if present, using Benson and Cummings' criteria and the Clinical Dementia Rating Scale, respectively. A trained research assistant completed telephone interviews to proxy informants for collateral information concerning cognition, behavior, and occupational or social function. Subjects' outpatient medical records were reviewed for documentation of problems with cognition. RESULTS: Twenty-six cases of dementia were identified. Of these 26, 17 (65%) (95% confidence interval, 44.3-82.8) were not documented in outpatient medical records; of 18 patients, 12 (67%) (95% confidence interval, 40.9-86.7) were not thought to have dementia by their physicians at the time of the office visit. Recognition and documentation rates increased with advancing stage of disease. CONCLUSION: Dementia is often unrecognized and undocumented in private practice settings. Arch Intern Med. 2000;160:2964-2968 PMID- 11041905 TI - Exercise and changes in health status in patients with ankylosing spondylitis. AB - BACKGROUND: Although exercise is a commonly recommended treatment for ankylosing spondylitis (AS), little is known about the effectiveness of unsupervised recreational and back exercises. We examined the effects of recreational exercise and back exercises on patient-reported pain severity, stiffness severity, and functional disability in a prospective longitudinal study of 220 patients with AS. METHODS: Participants provided information on exercise habits and health status every 6 months using mailed questionnaires (median follow-up, 4.5 years). Pain severity and stiffness severity were measured using visual analog scales, and functional disability was measured using the Health Assessment Questionnaire (HAQ) Disability Index. RESULTS: Among all patients, there were no associations between either the number of exercise minutes per week or the number of days of back exercise per week and short-term (6-month) changes in pain, stiffness, or HAQ Disability Index. However, among those who had AS for 15 years or less, pain scores were 0.18 points lower (on a scale of 0-3; P =.04), and stiffness scores were 6.4 points lower (on a scale of 0-100; P =.005) during periods with more than 200 minutes per week of exercise compared with periods with 0 to 30 minutes of exercise per week. Among those who had AS for more than 15 years, pain scores were 0.11 points lower (on a scale of 0-3; P =. 03), and HAQ Disability Indexes were 0.08 points lower (on a scale of 0-3; P<.001) during periods with 5 to 7 days per week of back exercise compared with periods when back exercises were not performed. Less intense levels of exercise were not associated with improvements in health status. CONCLUSIONS: Unsupervised recreational exercise improves pain and stiffness, and back exercise improves pain and function in patients with AS, but these effects differ with the duration of AS. Health status is improved when patients perform recreational exercise at least 30 minutes per day and back exercises at least 5 days per week. Arch Intern Med. 2000;160:2969-2975 PMID- 11041906 TI - Clinical prediction of acute aortic dissection. AB - BACKGROUND: Clinical criteria for aortic dissection are poorly defined. Thus, 35% of aortic dissections remain unsuspected in vivo, and 99% of suspected cases can be refuted. OBJECTIVE: To identify independent predictors of acute aortic dissection and create a prediction model for facilitated estimation of the individual risk of dissection. METHODS: Two hundred fifty patients with acute chest pain, back pain, or both; absence of an established differential diagnosis of the pain syndrome; and clinical suspicion of acute aortic dissection were evaluated for the presence of 26 clinical variables in a prospective, observational study. Multivariate analysis was performed to create a prediction model of aortic dissection. RESULTS: Aortic pain with immediate onset, a tearing or ripping character, or both; mediastinal widening, aortic widening, or both on chest radiography; and pulse differentials, blood pressure differentials, or both (P<.001 for all) were identified as independent predictors of acute aortic dissection. Probability of dissection was low with absence of all 3 variables (7%), intermediate with isolated findings of aortic pain or mediastinal widening (31% and 39%, respectively), and high with isolated pulse or blood pressure differentials or any combination of the 3 variables (> or = 83%). Accordingly, 4% of all dissections were assigned to the low-probability group, 19% to the intermediate-probability group, and 77% to the high-probability group of aortic dissection. CONCLUSIONS: Assessment of 3 clinical variables permitted identification of 96% of the acute aortic dissections and stratification into high-, intermediate-, and low-probability groupings of disease. With better selection for prompt diagnostic imaging, this prediction model can be used as an aid to improve patient care in aortic dissection. Arch Intern Med. 2000;160:2977 2982 PMID- 11041907 TI - Hyperhomocysteinemia is associated with the presence of retinopathy in type 2 diabetes mellitus: the Hoorn study. AB - BACKGROUND: Retinopathy is the leading cause of blindness among patients with type 2 diabetes mellitus (DM). Hyperhomocysteinemia is a recently recognized risk factor for cardiovascular disease, independent of established risk factors. OBJECTIVE: To study the association between the homocysteine level and retinopathy among subjects with and without DM. METHODS: We studied an age-, sex , and glucose tolerance-stratified random sample of a 50- to 75-year-old general white population in the Hoorn Study (N = 625). Retinal vascular changes (retinopathy) were assessed using ophthalmoscopy and/or fundus photography. Hyperhomocysteinemia was defined as a serum total homocysteine level greater than 16 micromol/L. RESULTS: The prevalence of retinopathy was 9.8% (28/285) in subjects with normal glucose tolerance, 11.8% (20/169) in those with impaired glucose tolerance, 9.4% (10/106) in those with newly diagnosed type 2 DM, and 32.3% (21/65) in those with known type 2 DM. The prevalence of retinopathy was 10.3% (39/380) in subjects without hypertension and 16.3% (40/245) in subjects with hypertension; it was 12.0% (64/534) in subjects with a serum total homocysteine level of 16 micromol/L or less and 16.5% (15/91) in those with a serum total homocysteine level of more than 16 micromol/L. After stratification for DM and adjustment for age, sex, glycosylated hemoglobin, and hypertension, the odds ratio (95% confidence interval) for the relation between retinopathy and hyperhomocysteinemia was 0.97 (95% confidence interval, 0.42-2.82) in patients without DM and 3.44 (95% confidence interval, 1.13-10.42) in patients with DM (P =.08 for interaction). CONCLUSION: The findings suggest that hyperhomocysteinemia may be a risk factor for retinopathy in patients with type 2 DM, but probably not in patients without DM. Arch Intern Med. 2000;160:2984-2990 PMID- 11041908 TI - Clinical factors that influence patients' desire for participation in decisions about illness. AB - BACKGROUND: Clinical practice often fails to optimize patient participation in decisions about serious illness. Prior studies are unclear about whether the type of decision and prior illness experience affect the patient's preferences for participation in decision making. Most studies of patient decision making have not addressed decisions about serious illness. OBJECTIVE: To determine whether the type of illness and nature of the decision predict the patient's preferences for involvement in making decisions. DESIGN: Study of randomly selected patients' responses to vignettes about cancer, acute myocardial infarction, and diabetes coupled with cross-sectional survey and chart review. SETTING: Outpatient Veterans Affairs medical clinic. PATIENTS: A total of 255 patients with a mean age of 63.2 years (95.2% male; 61.9% married). MAIN RESULTS: Patients wanted to share hypothesized major decisions with their physicians (mean score, 2.9; 1 = only physician, 5 = only patient) but wanted less involvement in hypothesized minor decisions (mean score, 2.5). Patients with recent severe heart disease (myocardial infarction, bypass surgery, angioplasty) wanted more involvement in decisions about acute myocardial infarction than did patients with stable angina or no heart disease; prior experience with diabetes did not affect decisions about diabetes. Factor analysis of the vignette items yielded 3 types of decisions that we consider to reflect major, minor, and patient behavior decisions. Mean scores were 2.9 for major decisions, 2.1 for minor decisions, and 2.7 for patient behavior decisions. CONCLUSIONS: Patients want to share in major decisions with their physicians but prefer to be less involved in minor decisions. For some illnesses, such as myocardial infarction, prior experience with the illness increases the patients' desire for participation in decision making. Arch Intern Med. 2000;160:2991-2996 PMID- 11041909 TI - Gastrointestinal tolerability of the selective cyclooxygenase-2 (COX-2) inhibitor rofecoxib compared with nonselective COX-1 and COX-2 inhibitors in osteoarthritis. AB - BACKGROUND: Most nonsteroidal anti-inflammatory drugs (NSAIDs) are nonselective cyclooxygenase (COX-1 and COX-2) inhibitors and are associated with a variety of upper gastrointestinal (GI) tract symptoms. The roles of COX-1 and COX-2 in the pathogenesis of these symptoms are unclear. To test whether COX-2 inhibition with rofecoxib would have greater GI tolerability than nonselective COX-1 and COX-2 inhibition, we compared the incidences of (1) treatment discontinuations for GI adverse events (AEs) and (2) prespecified dyspeptic-type GI AEs among patients with osteoarthritis treated with rofecoxib vs NSAIDs. METHODS: A prespecified, combined analysis of investigator-reported GI AEs in all 8 double-blind, randomized, phase 2b/3 osteoarthritis trials of rofecoxib was conducted. Patients included men and women with osteoarthritis (N = 5435); there was no upper age limit for entry. Treatments tested included rofecoxib, 12.5, 25, or 50 mg (combined), vs ibuprofen, diclofenac, or nabumetone (combined). Primary outcomes were the time (by survival analysis) to (1) treatment discontinuation due to GI AEs and (2) first reported dyspeptic-type GI AE. Between-treatment comparisons were made by log-rank test. RESULTS: The number of treatment discontinuations caused by GI AEs during 12 months was significantly lower (P=.02) with rofecoxib vs NSAIDs (8.2 vs 12.0 per 100 patient-years; relative risk, 0.70; 95% confidence interval, 0.52-0.94). The incidence of prespecified dyspeptic-type GI AEs during the first 6 months was significantly lower (P=.02) with rofecoxib vs NSAIDs (69.3 vs 85.2 per 100 patient-years; relative risk, 0.85; 95% confidence interval, 0.74 0.97). However, the difference between treatments in dyspeptic-type GI AEs was attenuated after 6 months. CONCLUSION: Rofecoxib was associated with a lower incidence of treatment discontinuations due to GI AEs over 12 months and a lower incidence of dyspeptic-type GI AEs over 6 months than treatment with nonselective COX inhibitors, or NSAIDs. Arch Intern Med. 2000;160:2998-3003 PMID- 11041910 TI - Troglitazone as an anti-inflammatory agent. PMID- 11041911 TI - Aspirin in diabetes: beware of retinopathy. PMID- 11041913 TI - NSAIDs, intestinal cell integrity, and bacterial translocation in chronic heart failure. PMID- 11041914 TI - beta-Blockers in congestive heart failure: is there a difference between classes? PMID- 11041916 TI - Additional monitoring tools to improve the quality of anticoagulation management. PMID- 11041917 TI - Regimes of quantum degeneracy in trapped 1D gases AB - We discuss the regimes of quantum degeneracy in a trapped 1D gas and obtain the diagram of states. Three regimes have been identified: the Bose-Einstein condensation (BEC) regimes of a true condensate and quasicondensate, and the regime of a trapped Tonks gas (gas of impenetrable bosons). The presence of a sharp crossover to the BEC regime requires extremely small interaction between particles. We discuss how to distinguish between true and quasicondensates in phase coherence experiments. PMID- 11041918 TI - Stochastic ballistic annihilation and coalescence AB - We study a class of stochastic ballistic annihilation and coalescence models with a binary velocity distribution in one dimension. We obtain an exact solution for the density which reveals a universal phase diagram for the asymptotic density decay. By universal we mean that all models in the class are described by a single phase diagram spanned by two reduced parameters. The phase diagram reveals four regimes, two of which contain the previously studied cases of ballistic annihilation. The two new phases are a direct consequence of the stochasticity. The solution is obtained through a matrix product approach and builds on properties of a q-deformed harmonic oscillator algebra. PMID- 11041919 TI - Deviations from linear theory for fluctuations below the supercritical primary bifurcation to electroconvection AB - Over two decades ago it was predicted that nonlinear interactions between thermally driven fluctuations in dissipative nonlinear nonequilibrium systems lead to deviations from mean-field theory. Here we report experimental observations of such deviations as a supercritical primary bifurcation is approached. We measured the mean-square director-angle fluctuations below the bifurcation to electroconvection of two different nematic liquid crystals. For epsilon(mf) identical withV2/V(2)(c,mf)-1 less, similar-0.1 ( V is the applied voltage) we find approximately |epsilon(mf)|(-gamma) with gamma given by linear theory (LT). Closer to the bifurcation there are deviations from LT with a smaller gamma and with V(2)(c)>V(2)(c,mf). PMID- 11041920 TI - No black-hole theorem in three-dimensional gravity AB - A common property of known black-hole solutions in (2+1)-dimensional gravity is that they require a negative cosmological constant. To explain this, it is shown in this Letter that a (2+1)-dimensional gravity theory which satisfies the dominant energy condition forbids the existence of a black hole. PMID- 11041921 TI - Gravitational wave bursts from cosmic strings AB - Cusps of cosmic strings emit strong beams of high-frequency gravitational waves (GW). As a consequence of these beams, the stochastic ensemble of gravitational waves generated by a cosmological network of oscillating loops is strongly non Gaussian, and includes occasional sharp bursts that stand above the rms GW background. These bursts might be detectable by the planned GW detectors LIGO/VIRGO and LISA for string tensions as small as G&mgr; approximately 10(-13). The GW bursts discussed here might be accompanied by gamma ray bursts. PMID- 11041922 TI - Universal torsion-induced interaction from large extra dimensions AB - We consider the Kaluza-Klein (KK) scenario in which only gravity exists in the bulk. Without the assumption of symmetric connection, the presence of brane fermions induces torsion. The result is a universal axial contact interaction that dominates those induced by KK gravitons. This enhancement arises from a large spin density on the brane. Using a global fit to Z-pole observables, we find the 3sigma bound on the scale of quantum gravity to be 28 TeV for n = 2. If Dirac or light sterile neutrinos are present, the data from SN1987A increase the bound to sqrt[n] M(S)>/=210 TeV. PMID- 11041923 TI - Verifiable model of neutrino masses from large extra dimensions AB - We propose a new scenario of neutrino masses with a Higgs triplet (xi(++),xi(+),xi(0)) in a theory of large extra dimensions. Lepton number violation in a distant brane acts as the source of a very small trilinear coupling of xi to the standard Higgs doublet in our brane. Small realistic Majorana neutrino masses are naturally obtained with the fundamental scale M(*) approximately O(1) TeV, foretelling the possible discovery of xi (m(xi) less, similarM(*)) at future colliders. Decays of xi(++) into same-sign dileptons are fixed by the neutrino mass matrix. Observation of &mgr;-e conversion in nuclei is predicted. PMID- 11041924 TI - Two-body random ensembles: from nuclear spectra to random polynomials AB - The two-body random ensemble for a many-body bosonic theory is mapped to a problem of random polynomials on the unit interval. In this way one can understand the predominance of 0(+) ground states, and analytic expressions can be derived for distributions of lowest eigenvalues, energy gaps, density of states, and so forth. Recently studied nuclear spectroscopic properties are addressed. PMID- 11041925 TI - Single atoms in an optical dipole trap: towards a deterministic source of cold atoms AB - We describe a simple experimental technique which allows us to store a small and deterministic number of neutral atoms in an optical dipole trap. The desired atom number is prepared in a magneto-optical trap overlapped with a single focused Nd:YAG laser beam. Dipole trap loading efficiency of 100% and storage times of about one minute have been achieved. We have also prepared atoms in a certain hyperfine state and demonstrated the feasibility of a state-selective detection via resonance fluorescence at the level of a few neutral atoms. A spin relaxation time of the polarized sample of 4.2+/-0.7 s has been measured. Possible applications are briefly discussed. PMID- 11041926 TI - Routes to nonsequential double ionization AB - A method is proposed for the calculation of the S matrix for many-electron processes in intense-laser atom physics, in close analogy to the strong-field approximation for one-electron processes. Given a scenario of how some process evolves, corresponding approximations to the classical action are made which allow for the evaluation of the quantum-mechanical S matrix. The method is applied to the distribution of the total electronic momentum in nonsequential double ionization, and the results are compared to recent measurements. Good agreement is obtained for neon for a rescattering scenario. There is no comparable agreement for helium and argon, and possible alternative scenarios are discussed. PMID- 11041928 TI - Quantum limits on optical resolution AB - We discuss the ultimate limit imposed by quantum fluctuations of light for resolution of fine details in optical images. For this purpose, we extend in the quantum domain the classical analysis of the object reconstruction, or superresolution, in terms of prolate spheroidal function basis. We derive the expression for ultimate resolution limit in the reconstructed object using an illumination of the full object plane by a multimode squeezed vacuum. We show that the gain in resolution using multimode squeezed light is maximum when the Shannon number of the imaging system is close to unity. PMID- 11041929 TI - Superradiant emission dynamics of an optically thin material sample in a short decay-time optical cavity AB - We report observations of optical superradiant emission and the atomic evolution it drives under conditions closely approximating those originally envisioned in the classic work of Dicke [Phys. Rev. 93, 99 (1954)]. Our experiment involves an optically thin solid sample in a short-lifetime optical cavity whose homogeneous coherence is cryogenically stabilized. Pulsed coherent excitation initiates superradiant emission which subsequently drives the sample to higher or lower states of coherence. Suppression of dephasing via cryogenics and propagation effects through use of an optically thin sample and cavity provides one of the clearest and cleanest examples of Dicke superradiance yet reported. PMID- 11041927 TI - Channel mixing effects in the dissociative recombination of H+3 with slow electrons AB - We discuss the low-energy dissociative recombination of H+3, which strongly influences the abundance of this ion in diffuse interstellar molecular clouds. The kinetic couplings between the ionization continuum and the dissociative ground state of H3 have been used as input to a two-dimensional wave packet calculation of dissociation dynamics. The cross section obtained for direct dissociative recombination is much smaller than the latest experimental results. However, a multichannel quantum defect treatment shows that an indirect mechanism via bound Rydberg states of H3 prevails for this process. PMID- 11041930 TI - Accuracy comparison of absolute optical frequency measurement between harmonic generation synthesis and a frequency-division femtosecond comb AB - Using an iodine-stabilized He-Ne laser as a transfer oscillator, we compare absolute measurements of the optical frequency from a traditional frequency synthesis chain based on harmonic generation and from the frequency division technique of an ultrawide bandwidth femtosecond frequency comb. The agreement between these two measurements, both linked to the Cs standard, is 220+/-770 Hz, yielding a measurement accuracy of 1.6x10(-12). We report 473 612 353 604.8+/-1.2 kHz as a preliminary updated value of the absolute frequency of the " f" component for the He-Ne laser international standard at 633 nm. PMID- 11041931 TI - Enhanced quantum correlations in bound higher-order solitons AB - Quantum effects in N-bound solitons can be drastically enhanced compared to the fundamental soliton. In particular, the spectral photon-number correlations are much stronger. The formation of spectral domains of almost perfect positive and negative correlations is predicted. Criteria that are based on the Cauchy-Schwarz inequality are used for studying nonclassical correlations. Interferences between different soliton components of the N-bound solution being responsible for the strong changes in the coherent amplitude may also be a mechanism for the strong correlations. The results offer novel possibilities of producing light with highly nonclassical properties. PMID- 11041932 TI - Bubbles interactions in the cahn-hilliard equation AB - We study the dynamics of bubbles in the one dimensional Cahn-Hilliard equation. For a gas of diluted bubbles we find ordinary differential equations describing their interaction which permits us to describe the ulterior dynamics of the system in very good agreement with numerical simulations. PMID- 11041933 TI - Global coupling with time delay in an array of semiconductor lasers AB - Synchronization due to a weak global coupling with time delay in a semiconductor laser array is investigated both in the cw and self-pulsing regimes. A generalized form of the Kuramoto phase equations is derived and discussed analytically. The time delay is shown to induce in-phase synchronization in all dynamical regimes. Another form of synchronization is found which leads to local extinction of self-pulsing in the array. PMID- 11041934 TI - Subcritical bifurcations and nonlinear balloons in faraday waves AB - Bicritical points at wave numbers k(b) larger than the critical wave numbers k(c) are found in parametric surface waves (Faraday waves) using both numerical simulations and nonlinear analysis. Because k(b)-k(c) is small, it is argued that subcritical bifurcations at k>k(b) can be easily observed in experiments. In the second part we present a generic argument predicting the existence of nonlinear states resembling a balloon outside the instability region. The prediction is confirmed in simulations and it is argued to apply to other systems with similar instability curves. PMID- 11041935 TI - Spontaneous pattern formation in an effectively one-dimensional dielectric barrier discharge system AB - In a dielectric-barrier discharge between diametrically opposite sides of a narrow tube, discharge filaments stabilize at regular intervals along the tube's length. Three types of periodic patterns are observed, as is a disordered state in which filaments fire at apparently random positions and times. Time-resolved current measurements indicate that for each spatial pattern, a particular number of discharge stages occur during the voltage half-cycle. A preliminary model of the pattern-formation dynamics is described, motivating further work on time resolved imaging and investigations of surface charge distributions. PMID- 11041936 TI - Head-tail instability caused by electron clouds in positron storage rings AB - In positron or proton storage rings with many closely spaced bunches, an electron cloud can build up in the vacuum chamber due to photoemission or secondary emission. We discuss the possibility of a single-bunch two-stream instability driven by this electron cloud. Depending on the strength of the beam-electron interaction, the chromaticity and the synchrotron oscillation frequency, this instability either resembles a linac beam breakup or a head-tail instability. We present computer simulations of the instabilities, and compare the simulation results with analytical estimates. PMID- 11041937 TI - First observation of self-amplified spontaneous emission in a free-electron laser at 109 nm wavelength AB - We present the first observation of self-amplified spontaneous emission (SASE) in a free-electron laser (FEL) in the vacuum ultraviolet regime at 109 nm wavelength (11 eV). The observed free-electron laser gain (approximately 3000) and the radiation characteristics, such as dependency on bunch charge, angular distribution, spectral width, and intensity fluctuations, are all consistent with the present models for SASE FELs. PMID- 11041938 TI - Exact invariants for a class of three-dimensional time-dependent classical hamiltonians AB - An exact invariant is derived for three-dimensional Hamiltonian systems of N particles confined within a general velocity-independent potential. The invariant is found to contain a time-dependent function f(2)(t), embodying a solution of a third-order differential equation whose coefficients depend on the explicitly known trajectories of the particle ensemble. Our result is applied to a one dimensional time-dependent nonlinear oscillator and to a system of Coulomb interacting particles in a time-dependent quadratic external potential. PMID- 11041939 TI - Reversed current structure in a Z-pinch plasma AB - The current profile of a Z-pinch plasma is investigated using a one-dimensional magnetohydrodynamic code. Simulation results reveal the formation of a reversed current profile, its propagation, and an ejection of plasma at boundary region, which have been observed in previous experiments. A new physical mechanism is proposed to account for such phenomena. The physical mechanism involves the propagation of a shock wave. It is found that a reversed current profile appears when a shock wave reflected at axis expands in a compressing plasma column. PMID- 11041940 TI - Bifurcation in viscoresistive MHD: the hartmann number and the reversed field pinch AB - A scaling approach to the simplest viscoresistive MHD model reveals that the Prandtl number acts only through the inertia term. When this term is negligible the dynamics is ruled by the Hartmann number H only. This occurs for the reversed field pinch dynamics as seen by numerical simulation of the model. When H is large the system is in a multiple helicity state. In the vicinity of H = 2500 the system displays temporal intermittency with laminar phases of quasi-single helicity (SH) type. For lower H's two basins of SH are shown to coexist. SH regimes are of interest because of their nonchaotic magnetic field. PMID- 11041941 TI - Direct observation of self-focusing with subdiffraction limited resolution using near-field scanning optical microscope AB - The self-focusing effect in As2S3 glass has been studied using a near-field scanning optical microscope. Optical images of fine features in the self-focused beam were directly measured at the self-focus with approximately 100 nm spatial resolution. Because of the unusually large nonlinear refractive index at 690 nm, filaments with minimum size of 0.3 &mgr;m were observed in a 1.6 mW beam propagating through the 1.7 &mgr;m thin film. A qualitative analysis of our experimental results is presented. We show that nonparaxiality is responsible for arresting self-focusing, as predicted by recent theories. PMID- 11041943 TI - Evidence for negative cross correlations in vibrational dephasing in liquids: isotropic raman-line shift and width phenomena in isotopic mixtures of N2 and O2 AB - To experimentally confirm the hypothesis of negative cross terms between different dephasing mechanisms, potentially leading to a broadening of the isotropic Raman line upon isotopic dilution, we present a detailed study on the concentration dependencies of all of the line shifts and widths in the four isotopic mixtures of type (15N14N)(x)-(14N2)(1-x), (15N2)(x)-(14N2)(1-x), (15N2)(x)-(15N14N)(1-x), and (16O2)(x)-(18O2)(1-x) near the normal boiling point of nitrogen at T = 77.35 K. A quite disparate behavior of the nitrogen oscillators compared to the oxygen oscillators was observed in respect to a change of their isotopic baths. PMID- 11041942 TI - Structure and spectroscopy of surface defects from scanning force microscopy: theoretical predictions AB - A possibility to study surface defects by combining noncontact scanning force microscopy (SFM) imaging with atomically resolved optical spectroscopy is demonstrated by modeling an impurity Cr3+ ion at the MgO(001) surface with a SFM tip. Using a combination of the atomistic simulation and the ab initio electronic structure calculations, we predict a topographic noncontact SFM image of the defect and show that its optical transitions can be either enhanced or suppressed depending on the tip atomistic structure and its position relative to the defect. These effects should allow identification of certain impurity species through competition between radiative and nonradiative transitions. PMID- 11041944 TI - Molecular structure of polystyrene at Air/Polymer and Solid/Polymer interfaces AB - IR-visible sum-frequency generation (SFG) spectroscopy has been used in a total internal reflection geometry to study the molecular structure of polystyrene (PS) at PS/sapphire and PS/air interfaces, simultaneously. The symmetric vibrational modes of the phenyl rings dominate the SFG spectra at the PS/air interface as compared to the antisymmetric vibrational modes at the PS/sapphire interface. This indicates approximately parallel orientation of the phenyl rings at the PS/air interface while nearly perpendicular orientation at the PS/sapphire interface, with respect to the surface normal. PMID- 11041945 TI - Critical exponents for random knots AB - The size of a zero-thickness (no excluded volume) nonphantom polymer ring is shown to scale with chain length N in the same way as the size of the excluded volume (self-avoiding) linear polymer, that is, as Nnu, where nu approximately 0.588. The consequences of this fact are examined, including the sizes of trivial and nontrivial knots. PMID- 11041946 TI - Vacancies in metals: from first-principles calculations to experimental data AB - We have revealed, and resolved, an apparent inability of density functional theory, within the local density and generalized gradient approximations, to describe vacancies in Al accurately and consistently. The shortcoming is due to electron correlation effects near electronic edges and we show how to correct for them. We find that the divacancy in Al is energetically unstable and we show that anharmonic atomic vibrations explain the non-Arrhenius temperature dependence of the vacancy concentration. PMID- 11041947 TI - Determination of the of rate cross slip of screw dislocations AB - The rate for cross slip of screw dislocations during annihilation of screw dipoles in copper is determined by molecular dynamics simulations. The temperature dependence of the rate is seen to obey an Arrhenius behavior in the investigated temperature range: 225-375 K. The activation energy and the effective attempt frequency can therefore be extracted from the simulations. The transition state energy for the annihilation process is calculated by identifying the transition state using the nudged elastic band path technique. The two activation energies agree very well, indicating that transition state theory is applicable for this type of process. PMID- 11041948 TI - Reflected shock experiments on the equation-of-state properties of liquid deuterium at 100-600 GPa (1-6 mbar) AB - New laser-driven shock experiments have been used to study the equation-of-state (EOS) properties of liquid deuterium. Reflected shocks are utilized to increase the shock pressure and to enhance the sensitivity to differences in compressibility. The results of these experiments differ substantially from the predictions of the Sesame EOS. EOS models showing large dissociation effects with much greater compressibility (up to a factor of 2) agree with the data. By use of independent techniques, this experiment offers the first confirmation of an earlier observation of enhanced compressibility in liquid deuterium. PMID- 11041949 TI - Phase transition in the n > 2 honeycomb O(n) model AB - We determine the phase diagram of the O(n) loop model on the honeycomb lattice, in particular, in the range n>2, by means of a transfer-matrix method. We find that, contrary to the prevailing expectation, there is a line of critical points in the range between n = 2 and infinity. This phase transition, which belongs to the three-state Potts universality class, is unphysical in terms of the O(n) spin model, but falls inside the physical region of the n-component corner-cubic model. It can also be interpreted in terms of the ordering of a system of soft particles with hexagonal symmetry. PMID- 11041950 TI - Detection of the high eigenmodes of spin diffusion in porous media AB - The dynamics of spin diffusion in a fluid is governed by the Torrey-Bloch equations, and the solution is often expressed mathematically in an eigenmode expansion. We report an experimental demonstration of the excitation and detection of a wide range of eigenmodes in porous media by exploring the inhomogeneous internal magnetic field in the pore space. The nodal character of the eigenfunctions of the high eigenmodes was clearly observed. The methodology of excitation and detection of the high eigenmodes may be used to better characterize pore geometry. PMID- 11041951 TI - Correlated electromigration of H in the switchable mirror YH(3-delta) AB - Electromigration of hydrogen in YH(3-delta) is studied by exploiting the H concentration dependence of the optical transmission of YH(3-delta). We find the effective valence Z* of H in YH(3-delta) to be negative. Its value is dominated by a huge wind-force-like term, i.e., Z* approximately K/rho, with K approximately -60 mOmega cm. This value is 3 orders of magnitude larger than typical for H in metals. In an Arrhenius plot, the ratio of hydrogen and electron fluxes extrapolates to unity at infinite temperature, suggesting a one-to-one correlation of hydrogen and electron hopping. We discuss our results in the light of strong electron correlation theories which predict each proton to bind two electrons in a sort of Zhang-Rice singlet. PMID- 11041952 TI - Phonons, rotons, and layer modes of liquid 4He in aerogel AB - We report the first observation of two-dimensional layer modes in both fully filled and partially filled aerogel. Using complementary high-energy resolution and high statistical precision neutron scattering instruments, and two different 87% porous aerogel samples, we show that the three-dimensional (3D) phonon-roton excitation energies and lifetimes of liquid 4He in aerogel are the same as in bulk 4He within current precision. The layer modes are the excitations that distinguish aerogel from the bulk rather than a difference in the 3D roton energy. PMID- 11041953 TI - GaAs(001) surface under conditions of low As pressure: evidence for a novel surface geometry AB - Using density-functional theory we identify a new low-energy structure for GaAs(001) in an As-poor environment. The discovered geometry is qualitatively different from the usual surface-dimer based reconstructions of III-V semiconductor (001) surfaces. The stability of the new structure, which has a c(8x2) periodicity, is explained in terms of bond saturation and favorable electrostatic interactions between surface atoms. Simulated scanning tunneling microscopy images are in good agreement with experimental data, and a low-energy electron diffraction analysis supports the theoretical prediction. PMID- 11041954 TI - Nonstandard roughness of terraced surfaces AB - We present a class of surfaces which are simultaneously flat and rough over the same range of lengths. They are self-affine, have well-defined roughness exponents, and consist of terraces of all sizes. The coexistence of flat and rough makes them respond to different external interactions with variable roughness. We demonstrate this for optical scattering (including x rays), two wetting situations, diffusion currents, and catalysis. A terraced Cu surface is a "self-assembled" experimental example, and designs for nano- and micromachined examples are presented. PMID- 11041955 TI - Derivation and validation of mesoscopic theories for diffusion of interacting molecules AB - A mesoscopic theory for diffusion of molecules interacting with a long-range potential is derived for Arrhenius microscopic dynamics. Gradient Monte Carlo simulations are presented on a one-dimensional lattice to assess the validity of the mesoscopic theory. Results are compared for Metropolis and Arrhenius microscopic dynamics. Non-Fickian behavior is demonstrated and it is shown that microscopic dynamics dictate the steady-state concentration profiles. PMID- 11041956 TI - Shear response of molecularly thin liquid films to an applied air stress AB - The shear response of molecularly thin liquid films on solid substrates when subjected to an applied air stress has been measured. The response corresponds to viscous friction while the same films sheared between two solid surfaces display static friction. These results show that molecularly thin liquid films partially confined by a single solid surface do not solidify as when confined between two solid surfaces. We are also able to observe several novel properties for liquid films on single solid surfaces not previously observed or expected. PMID- 11041957 TI - Interaction induced delocalization for two particles in a periodic potential AB - We consider two interacting particles evolving in a one-dimensional periodic structure embedded in a magnetic field. We show that the strong localization induced by the magnetic field for particular values of the flux per unit cell is destroyed as soon as the particles interact. We study the spectral and dynamical aspects of this transition. PMID- 11041958 TI - Optical conductivity of the half-filled hubbard chain AB - We combine well-controlled analytical and numerical methods to determine the optical conductivity of the one-dimensional Mott-Hubbard insulator at zero temperature. A dynamical density-matrix renormalization group method provides the entire absorption spectrum for all but very small coupling strengths. In this limit we calculate the conductivity analytically using exact field-theoretical methods. Above the Lieb-Wu gap the conductivity exhibits a characteristic square root increase. For small to moderate interactions, a sharp maximum occurs just above the gap. For larger interactions, another weak feature becomes visible around the middle of the absorption band. PMID- 11041959 TI - Observation of anomalous single-magnon scattering in half-metallic ferromagnets by chemical pressure control AB - The temperature variation of the resistivity rho and specific heat C have been measured for prototypical half-metallic ferromagnets, R0. 6Sr 0.4MnO3, by controlling the one-electron bandwidth W. We have found variations in the temperature scalings of rho from approximately T2 ( R = La, and Nd) to approximately T3 ( R = Sm), and have interpreted the T3 law in terms of the anomalous single-magnon scattering process in the half-metallic system. PMID- 11041960 TI - Evolution of quasiparticle charge in the fractional quantum hall regime AB - The charge of quasiparticles in a fractional quantum Hall (FQH) liquid, tunneling through a partly reflecting constriction with transmission t, was determined via shot noise measurements. In the nu = 1/3 FQH state, a charge smoothly evolving from e(*) = e/3 for t(1/3) congruent with 1 to e(*) = e for t(1/3)<<1 was determined, agreeing with chiral Luttinger liquid theory. In the nu = 2/5 FQH state the quasiparticle charge evolves smoothly from e(*) = e/5 at t(2/5) congruent with 1 to a maximum charge less than e(*) = e/3 at t(2/5)<<1. Thus it appears that quasiparticles with an approximate charge e/5 pass a barrier they see as almost opaque. PMID- 11041961 TI - Gap inhomogeneities and the density of states in disordered d-wave superconductors AB - We report on a numerical study of disorder effects in 2D d-wave BCS superconductors. We compare exact numerical solutions of the Bogoliubov-de Gennes (BdG) equations for the density of states rho(E) with the standard T-matrix approximation. Local suppression of the order parameter near impurity sites, which occurs in self-consistent solutions of the BdG equations, leads to apparent power-law behavior rho(E) approximately |E|(alpha) with nonuniversal alpha over an energy scale comparable to the single-impurity resonance energy Omega(0). We show that the novel effects arise from static spatial correlations between the order parameter and the impurity distribution. PMID- 11041962 TI - Details of disorder matter in 2D d-wave superconductors AB - Within numerically exact solutions of the Bogoliubov-de Gennes equations, we demonstrate that discrepancies between predicted low-energy quasiparticle properties in disordered 2D d-wave superconductors occur because of the unanticipated importance of disorder model details and normal state particle-hole symmetry. For the realistic case, which is best described by a binary alloy model without particle-hole symmetry, we predict density-of-state suppression below an energy scale which appears to be correlated with the corresponding single impurity resonance. PMID- 11041963 TI - Observation of a BCS spectral function in a conventional superconductor by photoelectron spectroscopy AB - We present high-resolution photoelectron spectra on the A15-type conventional superconductor V 3Si, where-for the first time-both singularities of the BCS density of states can be resolved by photoemission spectroscopy (PES). With a transition temperature of about T(c) approximately 17 K the gap Delta(gap) of this compound has a magnitude of approximately 5 meV. A measurement by PES on this small energy scale requires a very high energy resolution (DeltaE less, similar5 meV) and sample temperatures significantly below T(c). PMID- 11041964 TI - Self-duality in superconductor-insulator quantum phase transitions AB - It is argued that close to a Coulomb interacting quantum critical point the interaction between two vortices in a disordered superconducting thin film separated by a distance r changes from logarithmic in the mean-field region to 1/r in the region dominated by quantum critical fluctuations. This gives support to the charge-vortex duality picture of the observed reflection symmetry in the current-voltage characteristics on both sides of the transition. PMID- 11041965 TI - Resonances, instabilities, and structure selection of driven josephson lattice in layered superconductors AB - We investigate the dynamics of the Josephson vortex lattice in layered high- T(c) superconductors at high magnetic fields. It is shown that the average electric current depends on the lattice structure and is resonantly enhanced when the Josephson frequency matches the frequency of the plasma mode. We find the stability regions of a moving lattice. It is shown that a specific lattice structure at a given velocity is uniquely selected by the boundary conditions; at small velocities a periodic triangular lattice is stable and looses its stability at some critical velocity. At even higher velocities, a structure close to a rectangular lattice is restored. PMID- 11041966 TI - Chiral critical exponents of the triangular-lattice antiferromagnet CsMnBr3 as determined by polarized neutron scattering AB - The critical exponents gamma(c) = 0.84(7) of the chiral susceptibility above the Neel temperature, T(N), and beta(c) = 0. 44(2) of the average chirality below T(N) have been determined for the triangular-lattice antiferromagnet CsMnBr3 by means of polarized neutron scattering. These first experimental values of chiral critical exponents are in line with theoretical predictions and fulfill their scaling relation. The temperature at which the average chirality appears coincides with the spin-order transition temperature, T(N). PMID- 11041967 TI - Spin dynamics and orbital state in LaTiO3 AB - A neutron scattering study of the Mott-Hubbard insulator LaTiO3 ( T(N) = 132 K) reveals a spin wave spectrum that is well described by a nearest-neighbor superexchange constant J = 15.5 meV and a small Dzyaloshinskii-Moriya interaction ( D = 1.1 meV). The nearly isotropic spin wave spectrum is surprising in view of the absence of a static Jahn-Teller distortion that could quench the orbital angular momentum, and it may indicate strong orbital fluctuations. A resonant x ray scattering study has uncovered no evidence of orbital order in LaTiO3. PMID- 11041968 TI - Orbital liquid in three-dimensional mott insulator: LaTiO3 AB - We present a theory of spin and orbital states in Mott insulator LaTiO3. The spin orbital superexchange interaction between d(1)(t(2g)) ions in cubic crystal suffers from a pathological degeneracy of orbital states at the classical level. Quantum effects remove this degeneracy and result in the formation of the coherent ground state, in which the orbital moment of t(2g) level is fully quenched. We find a finite gap for orbital excitations. Such a disordered state of local degrees of freedom on unfrustrated, simple cubic lattice is highly unusual. Orbital liquid state naturally explains observed anomalies of LaTiO3. PMID- 11041969 TI - Charge ordering and polaron formation in the magnetoresistive oxide La0.7Ca0.3MnO3 AB - Neutron scattering has been used to study the nature of the ferromagnetic transition in a single crystal of the perovskite La0. 7Ca0.3MnO3. Diffuse scattering from lattice polarons develops as the Curie temperature is approached from below, along with short range polaron correlations that are consistent with stripe formation. Both the scattering due to the polaron correlations and the anomalous quasielastic component in the magnetic fluctuation spectrum maximize very close to T(C), in a manner remarkably similar to the resistivity, indicating that they have a common origin. PMID- 11041970 TI - Dispersion and symmetry of bound states in the shastry-sutherland model AB - Bound states made from two triplet excitations on the Shastry-Sutherland lattice are investigated. Based on the perturbative unitary transformation by flow equations quantitative properties like dispersions and qualitative properties like symmetries are determined. The high order results [up to (J2/J1)(14)] permit one to fix the parameters of SrCu2(BO3)(2) precisely: J1 = 6.16(10) meV, x J2/J1 = 0.603(3), J( perpendicular) = 1.3(2) meV. At the border of the magnetic Brillouin zone a general double degeneracy is derived. An unexpected instability in the triplet channel at x = 0.63 indicates a transition towards another phase. The possible nature of this phase is discussed. PMID- 11041971 TI - Anisotropy of domain wall resistance AB - The resistive effect of domain walls in FePd films with perpendicular anisotropy was studied experimentally as a function of field and temperature. The films were grown directly on MgO substrates, which induces an unusual virgin magnetic configuration composed of 60 nm wide parallel stripe domains. This allowed us to carry out the first measurements of the anisotropy of domain wall resistivity in the two configurations of current perpendicular and parallel to the walls. At 18 K, we find 8.2% and 1.3% for the domain wall magnetoresistance normalized to the wall width (8 nm) in these two respective configurations. These values are consistent with the predictions of Levy and Zhang. PMID- 11041972 TI - Negative refraction makes a perfect lens AB - With a conventional lens sharpness of the image is always limited by the wavelength of light. An unconventional alternative to a lens, a slab of negative refractive index material, has the power to focus all Fourier components of a 2D image, even those that do not propagate in a radiative manner. Such "superlenses" can be realized in the microwave band with current technology. Our simulations show that a version of the lens operating at the frequency of visible light can be realized in the form of a thin slab of silver. This optical version resolves objects only a few nanometers across. PMID- 11041973 TI - Vibrational energy transfer in a protein molecule. AB - Mode coupling in a protein molecule was studied by a molecular dynamics simulation of the intramolecular vibrational energy transfer in myoglobin at near zero temperature. It was found that the vibrational energy is transferred from a given normal mode to a very few number of selective normal modes. These modes are selected by the relation between their frequencies, like Fermi resonance, governed by the third order mode coupling term. It was also confirmed that the coupling coefficients had high correlation with how much the coupled modes geometrically overlapped with each other. PMID- 11041975 TI - Comment on "New conditions for a total neutrino conversion in a medium" PMID- 11041974 TI - Synchronization by irregular inactivation. AB - Many natural and technological systems have on/off switches. For instance, mitosis can be halted by biochemical switches which act through the phosphorylation state of a complex called mitosis promoting factor. If switching between the on and off states is periodic, chaos is observed over a substantial portion of the on/off time parameter plane. However, we have discovered that the chaotic state is fragile with respect to random fluctuations in the on time. In the presence of such fluctuations, two uncoupled copies of the system (e.g., two cells) controlled by the same switch rapidly synchronize. PMID- 11041976 TI - Chizhov and petcov reply: PMID- 11041977 TI - Comment on "Gravity waves, chaos, and spinning compact binaries" PMID- 11041978 TI - Comment on "Dramatic change of the magnetic response in LiV2O4: possible heavy fermion to itinerant d-metal Transition" PMID- 11041979 TI - Krimmel et al. reply: PMID- 11041980 TI - Comment on "Cuprate superconductivity: dependence of T(c) on the c-axis layering structure" PMID- 11041981 TI - Leggett replies: PMID- 11041982 TI - Non-fermi liquid angle resolved photoemission line shapes of li0. 9Mo6O17 PMID- 11041983 TI - Smith et al. reply PMID- 11041984 TI - High affinity binding of proteins HMG1 and HMG2 to semicatenated DNA loops. AB - BACKGROUND: Proteins HMG1 and HMG2 are two of the most abundant non histone proteins in the nucleus of mammalian cells, and contain a domain of homology with many proteins implicated in the control of development, such as the sex determination factor Sry and the Sox family of proteins. In vitro studies of interactions of HMG1/2 with DNA have shown that these proteins can bind to many unusual DNA structures, in particular to four-way junctions, with binding affinities of 10(7) to 10(9) M(-1). RESULTS: Here we show that HMG1 and HMG2 bind with a much higher affinity, at least 4 orders of magnitude higher, to a new structure, Form X, which consists of a DNA loop closed at its base by a semicatenated DNA junction, forming a DNA hemicatenane. The binding constant of HMG1 to Form X is higher than 5 x 10(12) M(-1), and the half-life of the complex is longer than one hour in vitro. CONCLUSIONS: Of all DNA structures described so far with which HMG1 and HMG2 interact, we have found that Form X, a DNA loop with a semicatenated DNA junction at its base, is the structure with the highest affinity by more than 4 orders of magnitude. This suggests that, if similar structures exist in the cell nucleus, one of the functions of these proteins might be linked to the remarkable property of DNA hemicatenanes to associate two distant regions of the genome in a stable but reversible manner. PMID- 11041985 TI - Cognitive neuroscience of actions: a whole is greater than the sum of its parts. AB - The purpose of the present issue of Brain and Cognition is to demonstrate that the strength of neuroscience is in its multidisciplinary approach to understand events. Here we are concerned with the domain of action recognition and production. Seven articles have been selected as representative studies of actions carried out in different areas of neuroscience such as neuropsychology, neurophysiology, and cognitive science. Although each addresses different questions within the field, the articles share a common neuroscientific knowledge, as can be readily gathered from the references cited in each article. PMID- 11041986 TI - Compatibility between observed and executed finger movements: comparing symbolic, spatial, and imitative cues. AB - Intuitively, one can assume that imitating a movement is an easier task than responding to a symbolic stimulus like a verbal instruction. Support for this suggestion can be found in neuropsychological research as well as in research on stimulus-response compatibility. However controlled experimental evidence for this assumption is still lacking. We used a stimulus-response compatibility paradigm to test the assumption. In a series of experiments, it was tested whether observed finger movements have a stronger influence on finger movement execution than a symbolic or spatial cue. In the first experiment, we compared symbolic cues with observed finger movements using an interference paradigm. Observing finger movements strongly influenced movement execution, irrespective of whether the finger movement was the relevant or the irrelevant stimulus dimension. In the second experiment, effects of observed finger movements and spatial finger cues were compared. The observed finger movement dominated the spatial finger cue. A reduction in the similarity of observed and executed action in the third experiment led to a decrease of the influence of observed finger movement, which demonstrates the crucial role of the imitative relation of observed and executed action for the described effects. The results are discussed in relation to recent models of stimulus-response compatibility. Neurocognitive support for the strong relationship between movement observation and movement execution is reported. PMID- 11041987 TI - Cognition in action: testing a model of limb apraxia. AB - Assessment of limb apraxia is still suffering from Liepmann's legacy and performance in gesture-processing tests is generally rendered by classifying patients' profile according to the classic clinical labels of ideomotor and ideational apraxia. At odds with other cognitive functions, interpretation of apraxia has suffered from a lack of a reliable model which does justice to its complexity. Recently such a model has been proposed (Rothi et al., 1991, 1997). In this article a modified version of this model is presented and predictions are made according to its functional architecture. Five different patterns of impairment of gesture processing are postulated. To validate the predicted performance profiles, 19 left-hemisphere-damaged patients were assessed by means of an ad hoc battery of four praxis tests. Four of the five predicted apraxia patterns were observed, the fifth being more equivocal. These results support the need to overcome the simplistic dichotomous view of apraxia and confirm the fruitfulness of a model of normal gesture processing in order to understand dissociations in apraxia. PMID- 11041988 TI - The role of the dynamic body schema in praxis: evidence from primary progressive apraxia. AB - On an influential model of limb praxis, ideomotor apraxia results from damage to stored gesture representations or disconnection of representations from sensory input or motor output (Heilman & Gonzalez Rothi, 1993; Gonzalez Rothi et al., 1991). We report data from a patient with progressive ideomotor limb apraxia which cannot be readily accommodated by this model. The patient, BG, is profoundly impaired in gesturing to command, to sight of object, and to imitation, but gestures nearly normally with tool in hand and recognizes gestures relatively well. In addition, performance is profoundly impaired on imitation of meaningless gestures and on tasks requiring spatiomotor transformations of body position information. We provide evidence that BG's apraxia is largely attributable to impairments external to the stored gesture system in procedures coding the dynamic positions of the body parts of self and others; that is, the body schema. We propose a model of a dynamic, interactive praxis system subserved by posterior parietal cortex in which stored representational elements, when present, provide "top-down" support to spatiomotor procedures computed on-line. In addition to accounting for BG's performance, this model accommodates a common pattern of ideomotor apraxia more readily than competing accounts. PMID- 11041989 TI - Perceiving human locomotion: priming effects in direction discrimination. AB - During the perception of biological motion, the available stimulus information is confined to a small number of lights attached to the major joints of a moving actor. Despite this drastic impoverishment of the stimulus, the human visual apparatus organizes the swarm of moving dots in a vivid percept of a human figure. In addition, observers effortlessly identify the action the figure is involved in. After a historical introduction and a short walk through the literature, data from a priming experiment are presented. In a serial two-choice reaction-time task, participants were presented with a point-light walker, facing either to the right or to the left and walking either forward or backward on a treadmill. Subjects had to identify the direction of articulatory movements. Reliable priming effects were established in consecutive trials, but these effects were tempered by the relation between priming and primed walker. The reaction time to a walker was shorter when the walker in the preceding trial moved in the same direction and was facing in the same direction. The findings are discussed in relation to recent data from neuropsychological case studies, neuroimaging, and single-cell recording. PMID- 11041990 TI - The role of semantic knowledge and working memory in everyday tasks. AB - We present a single case study of a patient, HG, who was severely impaired on routine everyday tasks, such as cleaning his teeth and preparing a cup of tea. We used the Action Coding System developed by Schwartz et al. (1991) to provide quantitative and qualitative measures of his performance in a number of experimental manipulations: (a) with task-congruent objects only, (b) with task congruent objects and semantic distractors, (c) with a set of written commands to follow, (d) when he was given one command at a time, (e) when he was shown how the task should be performed before starting himself, and (f) when the task was divided into smaller subgoals. In general, the majority of HG's errors were step omissions, perseverations, sequence errors, and semantic errors. These semantic errors are particularly interesting since HG was able to name, gesture to, and define all the objects when they were presented in isolation or in task-congruent arrays. We suggest that semantic errors may arise for a number of reasons: (1) impaired access from semantic memory to a network representing action schema, (2) degradation of stored schema, and (3) behavior that is abnormally driven by the goal, by preceding actions, or by salient objects rather than by an appropriate association between these elements in working memory. PMID- 11041991 TI - Task demands and limb apraxia in stroke. AB - The present study was designed to examine the frequency and severity of apraxia in patients with left- or right-hemisphere stroke in both pantomime and imitation conditions and to compare the frequency of apraxia in each stroke group across the three patterns of apraxia described in Roy's model (Roy, 1996). Ninety-nine stroke patients and 15 age-matched healthy adults performed eight transitive gestures to pantomime and to imitation. Gestural performance was quantified as accuracy on five performance dimensions; a composite score, an arithmetic combination of the five performance dimensions, was used as an index of the overall accuracy. Analyses revealed a comparable proportion of patients in each stroke group were classified as apraxic in the imitation condition, but a higher proportion of left stroke patients were apraxic in the pantomime condition. The severity of apraxia in each stroke group and the performance dimensions affected were, however, comparable. Analyses of the patterns of apraxia (pantomime alone, imitation alone or apraxia in both conditions) revealed a higher frequency of apraxia in both stroke groups for the pattern reflecting apraxia in both conditions, indicating that a disruption at the movement execution stage of gesture performance was most common. PMID- 11041992 TI - Neural representation for the perception of the intentionality of actions. AB - A novel population of cells is described, located in the anterior part of the superior temporal sulcus (STSa, sometimes called STPa) of the temporal lobe in the macaque monkey. These cells respond selectively to the sight of reaching but only when the agent performing the action is seen to be attending to the target position of the reaching. We describe how such conditional selectivity can be generated from the properties of distinct cell populations within STSa. One cell population responds selectively to faces, eye gaze, and body posture, and we argue that subsets of these cells code for the direction of attention of others. A second cell population is selectively responsive to limb movement in certain directions (e.g., responding to an arm movement to the left but not to an equivalent leg movement or vice versa). The responses of a subset of cells sensitive to limb movement are modulated by the direction of attention (indicated by head and body posture of the agent performing the action). We conclude that this combined analysis of direction of attention and body movements supports the detection of intentional actions. PMID- 11041993 TI - Replication of arboviruses in insect vectors. PMID- 11041994 TI - Experimental Elaphostrongylus cervi infection in sheep and goats. AB - The pathogenesis and migratory life cycle of Elaphostrongylus cervi were studied in four sheep and six goats killed and examined 6 days to 5 months after inoculation with infective third-stage larvae (L3). Detailed histological studies demonstrated that the L3 followed a porto-hepatic, and probably also a secondary lymphatic, migratory route from the abomasum and small intestine to the lungs, with subsequent spread via the general circulation to the central nervous system (CNS) and other tissues. In addition, the results suggested that haematogenously spread L3, arrested in arterial vessels outside the spinal cord, migrated into the cord along the spinal nerves. During migration, the L3 caused focal inflammation and necrosis in the organs and along the spinal nerve roots, and infarcts occurred in the myocardium, kidneys and CNS. Nematode development took place in the CNS. During development, there was a gradual die-off of nematodes and patent infections were not observed. However, in one animal many mature nematodes were demonstrated in the CNS. In the nervous system, the nematodes caused encephalomyelitis, focal traumatic encephalomalacia, gliosis, meningitis, choroiditis, radiculitis and perineuritis. Two goats and one sheep displayed long lasting paraparesis starting 6 weeks after inoculation. The signs apparently resulted from nematode-induced spinal nerve root lesions. From 19 weeks after inoculation the sheep also showed signs of severe brain disturbances due to traumatic and inflammatory lesions caused by adult E. cervi in the cerebral parenchyma. We conclude that E. cervi represents a potential cause of neurological disease in small ruminants grazing areas inhabited by red deer. This is the first report confirming the infectivity of E. cervi for domestic ruminants. PMID- 11041995 TI - Experimental transmission of porcine circovirus type 2 (PCV2) in weaned pigs: a sequential study. AB - Weaned specific pathogen-free pigs were inoculated intranasally with porcine circovirus type 2 (PCV2) and killed in groups of two or three animals at 6, 13, 20, 27 and 34 days post-inoculation (dpi), together with appropriate uninfected controls, for examination by histopathological, immunohistochemical (immunogold silver staining; IGSS), polymerase chain reaction (PCR) and viral isolation techniques. Serum samples were also collected for detection of antibodies. No major clinical signs were observed in infected pigs, and gross lesions were essentially limited to the lungs and lymph nodes of some of the animals. Histologically, no lesions were seen at 6 dpi, but bronchointerstitial pneumonia was invariably noted from 13 dpi onwards. Granulomatous inflammation, with or without intracytoplasmic inclusions, was present in lymphoid tissues (e.g. lymph nodes, thymus, spleen and tonsil) from day 20 onwards, being most severe at days 20 and 27 dpi. Liver inflammation was present at days 13, 20 and 27 dpi. Virus was demonstrated in the tissues by isolation and PCR methods throughout the experiment. PCV2 antigens were detected by IGSS in bronchial and bronchiolar epithelial cells, in mononuclear cells and multinucleated giant cells within inflammatory lesions, and in mononuclear cells of apparently normal tissues (e.glamina propria of the small intestine and the bronchus-associated lymphoid tissue). The lesions were consistent with those of postweaning multisystemic wasting syndrome (PMWS), although not all previously reported PMWS lesions were seen. PCV2 antibodies were detected in infected pigs from day 13 onwards. The results demonstrated widespread distribution of PCV2 after infection and persistence of the virus in vivo for at least 34 days. It would appear that PCV2 can induce PMWS lesions in weaned pigs in the absence of porcine parvovirus and other common swine pathogens. PMID- 11041996 TI - Cell differentiation and bone protein synthesis in the lungs of sheep with spontaneous calcinosis. AB - The lungs of three sheep with spontaneous enzootic calcinosis induced by the calcinogenic plant Nierembergia veitchii (Nv) were examined electron microscopically and immunohistochemically. The main ultrastructural changes were activation of fibroblasts and modified smooth muscle cells (SMCs) in the pulmonary interstitium, with an increase in extracellular matrix and precipitation of calcium, either in a laminated pattern or as amorphous aggregates. Macrophages and multinucleated giant cells, some with calcium crystals in the cytoplasm, were found in areas of increased extracellular matrix. Thickening and replication of the basal lamina of capillaries were prominent. The bone proteins osteocalcin, osteopontin and osteonectin were detected immunohistochemically in the cytoplasm of activated fibroblasts, in modified SMCs and in the extracellular matrix. It is suggested that 1,25-dihydroxyvitamin D(3)in Nv induces cellular differentiation and the synthesis of a calcifiable matrix. PMID- 11041997 TI - Cross-protection and antigen expression by chicken embryo-grown Pasteurella multocida in chickens. AB - The growth of Pasteurella multocida strain X-73 (serotype 1) and P-1059 (serotype 3) in vivo instead of in vitro resulted in the expression of additional antigens, as revealed by SDS-PAGE profile and western blotting of the outer membrane detergent-insoluble fraction (DIF-OM). Minor protein bands ranging from 58-138 kDa were expressed by both strains; major bands ranging from 35-64 and 35-43 kDa were expressed by X-73 and P-1059, respectively. Antigens at 35-39 kDa were dense and dominant. Treatment of DIF-OM from both strains with sodium periodate and proteinase K, but not with trypsin, reduced ELISA reactivity; this suggested that DIF-OM was composed of glycoprotein. Western blotting with heterologous antisera demonstrated antigenic cross-reactivity between strains X-73 and P-1059 grown in vivo. Inactivated vaccine prepared from X-73 grown in vivo protected chickens against challenge with X-73, P-1059 and strain P-1662 (serotype 4); a similar vaccine prepared from in-vitro culture, however, protected only against the homologous serotype. Moreover, antiserum against X-73 grown in vivo but not against the same strain grown in vitro gave considerable passive protection against challenge with the heterologous strains P-1059 and P-1662. PMID- 11041998 TI - Immunohistochemical characterization of lung lesions induced experimentally by Mycoplasma agalactiae and Mycoplasma bovis in goats. AB - Goats aged 3 months were inoculated with a recent isolate of Mycoplasma agalactiae (five animals) or Mycoplasma bovis (five animals) by a combined (intratracheal+intranasal) route. Two control goats were inoculated by the same route with sterile mycoplasma broth. Animals were killed 14 or 21 days after infection. At necropsy, tracheal and lung tissue was taken for pathological and immunohistochemical examination to determine changes in the lymphocyte subpopulations in the bronchus-associated lymphoid tissue (BALT). Consolidation of the lungs was not observed in any animal. M. agalactiae or M. bovis was recovered from the respiratory tract and lung of all but two infected animals. Both Mycoplasma spp. induced a moderate bronchointerstitial pneumonia, characterized by lymphoid hyperplasia of the BALT and infiltration of mononuclear cells into the alveolar walls. The predominant phagocytic cell in the pulmonary parenchyma and the airways was the macrophage. The main cellular type in the BALT was the CD3(+)T lymphocyte, and the ratio of CD4(+): CD8(+)cells was >1. It is likely that cellular immune mechanisms, through the activation of CD4(+)T lymphocytes, plays a prominent role in the acute and subacute phase of these infections. PMID- 11041999 TI - The histology and growth kinetics of canine renal oncocytoma. AB - A renal oncocytoma was diagnosed in a 10-year-old English springer spaniel with a 5-month history of anorexia, vomiting and weight loss. The tumour, which was localized to the kidney, was treated by simple nephrectomy and the dog made a full recovery. Histologically, the tumour consisted of cells with abundant eosinophilic cytoplasm forming acini and alveolar nests set within a loose fibrovascular stroma. The results of cell kinetic studies (AgNOR score 2.68, PCNA index 5.2%) were comparable with findings reported for benign human renal tumours. This appears to be the first reported case of renal oncocytoma in a dog. PMID- 11042000 TI - Amoebosis in the flat-shelled spider tortoise (Acinixys planicauda). AB - Amoebosis is one of the most common protozoal diseases of reptiles, but has rarely been reported in tortoises. Six cases of amoebosis were identified in flat shelled spider tortoises immediately after their importation from Madagascar to Japan. Necropsy revealed an extensive green pseudomembrane of ulceration along the length of the thickened wall of the colon, and disseminated green foci in the liver. Histologically, the colonic wall was severely ulcerated and covered with a fibrinonecrotic pseudomembrane, and many amoebic trophozoites were seen in the submucosa. Multifocal necrosis with intralesional amoebic trophozoites corresponded to the disseminated green foci in the liver. Amoebic trophozoites invaded many blood vessels in the colon and liver. All tortoises exhibited severe colonic lesions, but the severity of the hepatic lesions varied. These findings suggest that amoebic colitis was the primary lesion with spread of amoebae to the liver via the portal system. PMID- 11042001 TI - Distribution of porcine parvovirus in porcine circovirus 2-infected pigs with postweaning multisystemic wasting syndrome as shown by in-situ hybridization. AB - In-situ hybridization with a nonradioactive digoxigenin-labelled probe was used to study the distribution of porcine parvovirus (PPV) in formalin-fixed paraffin wax-embedded tissues from 10 porcine circovirus 2 (PCV2)-infected weaned pigs with naturally occurring postweaning multisystemic wasting syndrome (PMWS). A 226 base pair DNA fragment from a VP2 structural gene was generated by polymerase chain reaction (PCR) and used as a probe. Lymph node, spleen, thymus and tonsil were positive by PCR, demonstrating the presence of PPV DNA in the tissue samples from four of 10 pigs tested. PPV nucleic acid was also detected consistently in lymph node, spleen, thymus and tonsil by in-situ hybridization. Detection of PPV DNA from PCV2-infected pigs with PMWS suggests that PPV also plays a role in the pathogenesis of PMWS. PMID- 11042004 TI - Editorial PMID- 11042002 TI - Canine extracutaneous mast-cell tumours consisting of connective tissue mast cells. AB - Two cases of canine extracutaneous mast-cell tumours were encountered, originating from the mucosa of either the oral cavity or the small intestine. The dogs had no neoplasms in the skin. Immunohistochemical and ultrastructural studies demonstrated that the neoplastic cells had the features of connective tissue mast cells. It would seem, therefore, that at least some extracutaneous forms of the neoplasm originate from connective tissue mast cells. Heparin was a useful cytological marker to diagnose this type of mast-cell tumour. PMID- 11042005 TI - Adrenocortical function in a new world primate, the marmoset monkey, Callithrix jacchus. AB - The function of the adrenal cortex of the marmoset monkey Callithrix jacchus has been investigated. In common with other New World primates, these animals seem to be glucocorticoid resistant. Blood and adrenal glands were obtained from male and female animals under terminal pentobarbitone anesthesia. Dispersed adrenal cell preparations were obtained by treatment with collagenase and incubated with ACTH(1-24), (0.1-1000 nM) angiotensin II (0.1-1000 nM), dibutyryl cyclic AMP (dbcAMP; 30-1000 microM), and forskolin (FSK; 1-30 microM). Plasma cortisol levels (2113 +/- 449 ng/ml male; 3858 +/- 429 ng/ml female) were found to be 10- to 20-fold higher than those quoted for Old World primates and man. The cell preparations showed no significant response to any dose of ACTH tested (0.1-1000 nM), although addition of exogenous precursor (22R-hydroxycholesterol, 2.5 microM) resulted in an increased yield of cortisol and aldosterone. Cyclic AMP production was increased in response to forskolin (1-30 microM) but not ACTH(1 24) (1-1000 nM). In addition, dose-related responses to angiotensin II (maximal stimulation of 316 +/- 49% basal aldosterone at 100 nM angiotensin II), dbcAMP (maximal stimulation of 449 +/- 24% basal cortisol at 300 microM dbcAMP), and forskolin (maximal stimulation of 394 +/- 31% basal cortisol at 10 microM FSK) were obtained. The lack of a response in vitro to ACTH in C. jacchus cannot, therefore, be attributed either to general failure of the cells or to defects in postreceptor signaling mechanisms. The results suggest that there is a reduction in adrenal ACTH receptor number or affinity, with a high basal production rate in vivo maintaining the elevated plasma cortisol levels. PMID- 11042006 TI - Development of cholecystokinin and pancreatic polypeptide endocrine systems during the larval stage of Japanese flounder, Paralichthys olivaceus. AB - To understand the developmental process of the endocrine system, which controls the pancreatic exocrine function in Japanese flounder, Paralichthys olivaceus, the expression patterns of cholecystokinin (CCK) and pancreatic polypeptide (PP) during the larval stage were analyzed by immunohistochemistry and in situ hybridization. Expression of CCK in the intestinal epithelia started at 2 days posthatching (dph), 1 day prior to the first feeding. Endocrine pancreatic cells containing insulin were present in the pancreas primodium at hatching, and these endocrine cells formed an islet at 3 dph and developed into the principle islet neighboring gallbladder at 20 dph. However, PP cells were not contained in the principle islet even after metamorphosis. PP cells were contained in the accessory islets differentiated around the proximal part of the pyloric appendages at 30 dph (early metamorphosis stage). Therefore, we infer that the stimulative regulation of pancreatic enzyme secretion by CCK starts to function at the first feeding, whereas the restrictive regulation by PP develops about 1 month later in flounder larvae. In addition, we observed that CCK immunoreactive cells appeared in the accessory islets at 30 dph, similar to PP cells, even though CCK mRNA expression could not be detected in cells from the islets. This indicates the possibility that a peptide that is cross-reacted with the CCK antibody, i.e., gastrin, is synthesized in the flounder islets. PMID- 11042007 TI - Chromogranin A-immunoreactive cells in the olfactory system of anuran amphibians. AB - Chromogranin A (CgA) is a member of the granin family of acidic proteins that are present in the secretory granules of many endocrine and neuroendocrine cells. The specific function(s) of these proteins is not known, but they seem to be the precursors of biologically active peptides, and they may act as helper proteins in the sorting and packaging of peptide hormones and neuropeptides. Using indirect immunohistochemistry, we have found CgA immunoreactivity in the primary olfactory epithelia, the vomeronasal epithelia, the olfactory nerves, and the olfactory bulbs of tadpoles of the American toad, Bufo americanus, and the green frog, Rana clamitans. CgA immunoreactivity was present in the early stages of larval development in toads but was not detected in toad tadpoles after the hindlimb buds formed or in toadlets or adults. In green frog tadpoles, CgA immunoreactive cells were found in pre- and prometamorphic stages but not in late climax. CgA immunoreactivity was also absent in froglets, but it was detected in the vomeronasal epithelium but not the olfactory epithelium of adult green frogs. PMID- 11042008 TI - Pharmacological adrenalectomy with mitotane. AB - The potential of mitotane (ortho, para'-DDD, commonly used to treat adrenal carcinomas in humans and dogs) was investigated as an alternative to surgical adrenalectomy in birds, salamanders, and lizards. House sparrows (Passer domesticus) were injected twice daily with vehicle or one of two doses of mitotane (225 or 450 mg/kg), and basal and stress-induced levels of corticosterone (CORT) were measured 3 and 5 days after injections. Mitotane reduced basal CORT levels to nondetectable and abolished stress-induced CORT increases by the 3rd day of treatment. In another study, a single injection of mitotane was effective in lowering endogenous CORT levels 36 h later, but levels had apparently recovered by 10 days after the injection. Mitotane did not effect testicular weights and had no detectable effect on testosterone levels. In contrast to its effects on house sparrows, mitotane did not lower endogenous CORT levels in either tiger salamanders (Ambystoma tigrinum) or tree lizards (Urosaurus ornatus), even at doses much higher than those used in house sparrows. PMID- 11042009 TI - The presence of high-affinity, low-capacity estradiol-17beta binding in rainbow trout scale indicates a possible endocrine route for the regulation of scale resorption. AB - High-affinity, low-capacity estradiol-17beta (E(2)) binding is present in rainbow trout scale. The K(d) and B(max) of the scale E(2) binding are similar to those of the liver E(2) receptor (K(d) is 1.6 +/- 0.1 and 1.4 +/- 0.1 nM, and B(max) is 9.1 +/- 1.2 and 23. 1 +/- 2.2 fmol x mg protein(-1), for scale and liver, respectively), but different from those of the high-affinity, low-capacity E(2) binding in plasma (K(d) is 4.0 +/- 0.4 nM and B(max) is 625.4 +/- 63. 1 fmol x mg protein(-1)). The E(2) binding in scale was displaced by testosterone, but not by diethylstilbestrol. Hence, the ligand binding specificity is different from that of the previously characterized liver E(2) receptor, where E(2) is displaced by diethylstilbestrol, but not by testosterone. The putative scale E(2) receptor thus appears to bind both E(2) and testosterone, and it is proposed that the increased scale resorption observed during sexual maturation in both sexes of several salmonid species may be mediated by this receptor. No high-affinity, low capacity E(2) binding could be detected in rainbow trout gill or skin. PMID- 11042010 TI - Gonadotropin-releasing hormone-immunoreactive neurons and associated nicotinamide adenine dinucleotide phosphate-diaphorase-positive neurons in the brain of a teleost, Rhodeus amarus. AB - Using combined nicotinamide adenine dinucleotide phosphate-diaphorase (NADPHd) histochemistry and salmon gonadotropin-releasing hormone (sGnRH) immunocytochemistry, it is reported for the first time that possible potential contacts occur between the nitric oxide (NO)- and the GnRH-containing neurons in the brain of a freshwater teleost, Rhodeus amarus. GnRH-immunoreactive (ir) neurons were observed in the olfactory nerve (OLN), olfactory bulb (OB), medial olfactory tract (MOT), ventral telencephalon (VT), nucleus preopticus periventricularis (NPP), nucleus lateralis tuberis (NLT), and midbrain tegmentum (MT). Although NADPHd neurons were widely distributed in the brain, only those having an association with GnRH-ir neurons are described. Based on the nature of the association between the GnRH and the NADPHd neurons, the former were classified into three types. The Type I GnRH neurons were characterized by the presence of NADPHd-positive granules in the perikarya and processes and occurred in the OLN, OB, MOT, and VT. The Type II GnRH neurons, having soma-soma or soma process contacts with the NADPHd neurons, were restricted to the MT; the long processes of NADPHd cells crossed over either the perikarya or the thick processes of GnRH cells. However, the Type III GnRH neurons, found in the NPP and NLT, did not show direct contact, but a few NADPHd fibers were present in the vicinity. The terminal-soma contacts in the olfactory system and the VT and the soma-soma contacts in the MT represent the sites of possible potential contacts indicating a direct NO involvement in GnRH function, although NO action by diffusion remains possible. NO may influence the NPP and NLT GnRH cells by diffusion only, since a direct contact was not observed. PMID- 11042011 TI - Short-term stress increases testosterone secretion from testes in male domestic fowl. AB - Prolonged stress inhibits the hypothalamus-pituitary-gonadal (HPG) axis and reduces plasma testosterone (T). However, enhanced secretion of luteinizing hormone (LH) and T has been documented during the initial stages of acute stress in mammals. This study assayed the effect of short-term stress on plasma T and corticosterone (B) in juvenile, pubertal, and adult White Leghorn cockerels. Stress was induced by brief physical restraint of caged juvenile (7 weeks), pubertal (17 weeks), and adult (40 weeks) cockerels, as well as 40-week-old adults reared together in a room lined with wood shavings (group reared). Blood was sampled immediately before restraint (0 time), at the end of a 10-min restraint period, and at 30, 60, and 180 min after 0 time. Restraint resulted in an initial increase in plasma T in all groups, along with a rise in B. Whereas B generally reached its peak level at the end of the restraining period, T peaked 20 min later. The maximum increase of T and B relative to prestress levels (T and B ratios) was similar in all groups, with median T ratio reaching 1.25-1. 5-about half that of the B ratio. Thus, the extent of T and B response to short-term stress was not influenced by basal levels of T, which were highest in adults, and basal levels of B, which were higher in caged adults than in group-reared adults. Injection of ACTH did not induce a greater increase in plasma T than in sham injected controls. Further, the elevation of T in response to stress was extinguished in castrated adults, indicating that T is secreted from the testes rather than the adrenals in response to stress. When the same regime of blood sampling was applied to adults not subjected to restraint, the T ratio rose by up to 11 times. It can therefore be stipulated that T response depends on the type of stress applied, a factor that should be considered when investigating androgen levels in plasma. PMID- 11042012 TI - Pituitary levels of three forms of GnRH in the male European sea bass (Dicentrarchus labrax, L.) during sex differentiation and first spawning season. AB - In the present study, levels of three GnRH forms [seabream GnRH (sbGnRH), chicken GnRH-II (cGnRH-II), and salmon GnRH (sGnRH)] were analyzed in the pituitary of male sea bass during sex differentiation and the first spawning season. Plasma levels of gonadotropin (GTH-2), testosterone (T), and 11-ketotestosterone (11-KT) were determined during the same periods. All GnRH forms were present in the pituitary. sbGnRH levels were 9-fold higher than cGnRH-II and 17-fold higher than sGnRH levels. The highest GnRHs levels were detected in November 1995, when fish were 9 months old and when the gonads started to differentiate. Levels of the three forms decreased and remained low during the first spawning season, with the exception of sbGnRH, which showed a significant increase in November 1996. Plasma GTH-2 levels were lowest in November 1995, later increasing 2.5 times during the next months. During the first spawning season, plasma GTH-2 levels peaked in December 1996, 1 month after the peak of sbGnRH. During sex differentiation, plasma T levels were high in November 1995 but decreased over the next months, while levels of 11-KT remained low and unchanged. During the first spawning season, both steroids peaked in January 1997. These results suggest a possible role for all three GnRH forms in achieving gonadal differentiation, while sbGnRH may be the most relevant form in the regulation of the first spawning season in male sea bass. Moreover, GTH-2 and 11-KT may play important roles in gonadal maturation, since plasma GTH-2 and 11-KT levels were high throughout the period of spermiation. PMID- 11042013 TI - Biochemical and morphological diversity among folliculo-stellate cells of the mink (Mustela vison) anterior pituitary. AB - The folliculo-stellate (FS) cells are agranular cells of the anterior pituitary whose origin and function are still a matter of debate. This study examined the presence, topography, and morphological characteristics of FS cells in the mink anterior pituitary throughout the annual reproductive cycle. The S-100 protein was used as a FS cell marker. Immunoperoxidase labeling on tissue sections demonstrated the presence of two types of S-100 positive cells. Type 1 cells were stellate-shaped cells whose nuclei were localized near the center of pituitary follicles. In this type, S-100 labeling was strong in anterior pituitary sections obtained during spring, a period characterized by high prolactin pituitary content and low gonadotropin pituitary content. Type 2 cells were rounded cells occupying the periphery of the follicles. During periods of low prolactin pituitary content and high gonadotropin anterior content the type 2 S-100 positive cells formed aggregates of several cells. The total number of S-100 positive cells was constant during these two periods of the annual reproductive cycle, suggesting that type 1 and type 2 may reflect different morphological and physiological states of the same cell. Of the two subunits, alpha and beta, that, combined, form three different dimeric S-100 proteins, mink FS cells expressed mostly the beta subunit. FS cells also expressed the glial fibrillary acidic protein (GFAP). In culture, 8 +/- 3% of anterior pituitary cells were S-100 positive. Cultured S-100 cells were elongated, polygonal, or rounded. The S-100 labeling accumulated in the cytoplasm around and within the nucleus, whereas it was weak in pseudopods and large cytoplasmic vacuoles. The presence of pseudopods suggests that cultured FS cells could migrate. The vacuoles may be related to the phagocytic activity ascribed to these cells. Some FS cells presented membrane blebbing and peripheral vesicles that were immunopositive for S-100 and that may indicate a secretory activity. Cultured FS cells possessed actin filaments organized as a peripheral network; a few actin cables were also observed running across the cytoplasm. Pseudopods depicted a highly organized actin network. The microtubules of FS cells expanded throughout the cytoplasm. The intermediate filaments expressed by cultured FS cells were GFAP and vimentin. GFAP labeling was punctate and vimentin was organized as filaments. All cultured S-100 cells were positive for vimentin, suggesting a mesenchymal origin for the cells, and all cultured S-100 positive cells were positive for GFAP, suggesting a neuroectodermal origin. In conclusion, S-100 positive cells are heterogeneous with respect to cell shape and expression of S-100 subunits in the mink anterior pituitary. The presence of morphologically different S-100 positive cells is modified in accordance with the endocrine status of the animal, suggesting that FS cells may be involved in the modulation of the anterior pituitary endocrine activity in the mink. PMID- 11042014 TI - The effects of photoperiod and feeding on the diurnal rhythm of circulating thyroid hormones in the red drum, Sciaenops ocellatus. AB - Available data in cyprinid and salmonid species indicate that nutrient intake sustains thyroidal rhythmicity and that time of feeding may influence the amplitude, but not the phase, of diurnal thyroid hormone cycles. Several experiments were conducted to characterize the nature of thyroidal rhythmicity in a more derived perciform teleost, the red drum. These studies were designed to test the following hypotheses: (1) that feeding time will alter the amplitude of the thyroid hormone rhythm without altering its phase and (2) that food deprivation will diminish the amplitude of the thyroid hormone rhythm. Circulating T(4) levels in this species exhibit high-amplitude diurnal rhythms, whereas circulating T(3) levels fluctuate within a more narrow range. Fish were reared under a 12L:12D photoperiod and fed 5% body weight once daily either at dawn or at dusk. Feeding time had no discernible effect on the phase of the T(4) cycle, but altered the amplitude of the cycle. Dawn-fed fish had significantly greater mean peak levels of T(4) than dusk-fed fish, although there was no difference in daily mean levels in both groups of fish. When red drum were deprived of food, significant declines in plasma glucose, HSI, and liver glycogen content occurred within 3 days. When red drum were sampled once per day after 3, 7, or 11 days of food deprivation there were no consistent changes in circulating T(4) and T(3) levels compared to those of fed controls. However, significant declines in circulating T(4) and T(3) levels in response to food deprivation were detected with a diurnal sampling protocol. Within 3 days of food deprivation, T(4) levels were significantly reduced compared to those in fed controls and not significantly different from T(4) levels after 10 days of food deprivation. T(3) levels exhibited a stepwise decline in circulating levels during food deprivation. These data indicate that both feeding time and nutrient status exert their effects on thyroid hormone rhythms by modifying the amplitude of these cycles. These data also underscore the importance of incorporating a consideration of endocrine rhythmicity into sampling protocols. PMID- 11042015 TI - Dietary protein restriction stress in the domestic chicken (Gallus gallus domesticus) induces remodeling of adrenal steroidogenic tissue that supports hyperfunction. AB - The stress of dietary protein restriction in the immature domestic turkey (Meleagris gallopavo) induces adrenal steroidogenic hypofunction that is associated with an alteration in the proportion of density-dependent subpopulations of steroidogenic cells within the adrenal gland. In contrast, when imposed on immature chickens, this nutritional stressor induces long-term enhancement of adrenal steroidogenic function. However, whether this alteration in function is accompanied by a remodeling of chicken adrenal steroidogenic tissue as in the turkey is not known. To address this question, immature cockerels (2 weeks old) were fed established isocaloric synthetic diets containing either 20% (control) or 8% (restriction) soy protein for 4 weeks. Adrenal glands were processed for the isolation of defined, density-separable, adrenal steroidogenic cell subpopulations: three low-density adrenal steroidogenic cell subpopulations [LDAC-1 (rho = 1.0285-1.0430 g/ml), LDAC-2 (rho = 1. 0430-1.0485 g/ml), and LDAC-3 (rho = 1.0485-1.0500 g/ml)] and one high density subpopulation [HDAC (rho = 1.0510-1.0840 g/ml)]. The steroidogenic function of these cell subpopulations was assessed. Protein restriction consistently, but differentially, enhanced maximal ACTH-induced corticosterone production by the subpopulations: values of LDAC-1, -2, and -3 and HDAC from protein restricted birds were, respectively, 116, 43, 33, and 20% greater than those of corresponding cell subpopulations from control birds. However, it had contrasting influences on maximal ACTH-induced aldosterone production by the cell subpopulations. Whereas the value of LDAC-1 from protein-restricted birds was 70% greater than that from control birds, the values for LDAC-2 and -3 were not different from those of the control, and the value for HDAC was 22% less than that of the control. Protein restriction also altered the cell subpopulation composition of the adrenal gland: compared to control, it increased the proportion of LDAC-1 by 46% and decreased the proportion of LDAC-3 and HDAC by 34 and 20%, respectively. Thus, dietary protein restriction increased the proportion of cells (i.e., LDAC-1) having the greatest enhancement in corticosteroid production. This pattern of remodeling of chicken adrenal steroidogenic tissue in response to dietary protein restriction contrasts sharply with the pattern that occurs in another galliform species, the domestic turkey. PMID- 11042016 TI - High blood cortisol levels and low cortisol receptor affinity: is the chub, Leuciscus cephalus, a cortisol-resistant teleost? AB - In contrast to the relatively minor intra- and interspecies differences in blood cortisol levels reported for salmonid species, there is a more pronounced distinction between cortisol levels among the Salmonidae and the Cyprinidae, with both basal and stress-induced cortisol levels markedly higher in the latter. This study shows that in the chub, Leuciscus cephalus, a widely distributed European cyprinid, mean blood cortisol levels during stress (1500 ng mL(-1)) exceeded those reported for most other species of fish and, even in unstressed chub, cortisol levels (50-100 ng mL(-1)) were within the range known to cause immunosuppression, growth retardation, and reproductive dysfunction in salmonid fish. The chub appears to be atypical only with respect to plasma cortisol levels; the levels of plasma glucose and plasma lactate in unstressed and stressed chub are similar to those reported for other species. Plasma levels of 11-ketotestosterone in males and 17beta-estradiol in females are lower than those reported for salmonids but similar to those reported for other cyprinid species and display clear stress-induced reduction. Comparative analysis of the binding characteristics of the trout and chub gill cortisol receptor revealed that the total number of binding sites in gill tissue for each species was similar (B(max); approximately 50-100 fmol mg(-1) protein). However, the affinity of the binding site for cortisol displayed an eightfold difference between the species (rainbow trout: K(d) approximately 6 nM; chub: K(d) approximately 50 nM). Therefore, the potentially adverse effects of high circulating levels of cortisol found both at rest and under conditions of stress in chub may be offset by the lower affinity of the cortisol receptor, rather than the abundance of target tissue receptor sites. This strategy is similar to that reported for some glucocorticoid-resistant rodents and New World primates. PMID- 11042017 TI - Baseline corticosterone peaks in shorebirds with maximal energy stores for migration: a general preparatory mechanism for rapid behavioral and metabolic transitions? AB - In captive red knots (Calidris canutus, Scolopacidae) showing a regulated body mass increase of 50% related to their migration from temperate staging sites to tundra breeding grounds, plasma corticosterone concentrations increased from less than 10 ng. ml(-1) to levels as high as 30 ng. ml(-1) when the energy storage for migration was complete. These birds did not fly, but concentrations dropped to very low levels (<5 ng. ml(-1)) as soon as the birds started their voluntary fasts to the low body masses preceding the early wing and body molts normally occurring after an unsuccessful breeding season. As the elevated levels of corticosterone are associated with stable body mass rather than with the preceding increase or subsequent decrease, it is suggested that a major role of corticosterone during the final stages just before departure may be to prepare birds for long-distance flights. Birds heading into the Arctic to breed face potentially arduous flights into unpredictable environmental and social conditions. Activation of the hypothalamic-pituitary-adrenal axis, as measured by elevated levels of corticosterone, may induce the suite of behavioral and metabolic changes necessary to negotiate these challenges successfully. PMID- 11042018 TI - Unexpected and unexplained phenotypes in transgenic models. PMID- 11042019 TI - Comparison of STAT5 mRNA levels in GH-treated male and female rats analysed by a solution hybridization assay. AB - In this study we describe the development of a RNA:RNA solution hybridization RNase protection assay to quantify STAT5 mRNA in total RNA extracts from rat tissues. The assay is sensitive and reproducible. We quantified STAT5 mRNA levels in liver and thymus lymphocytes from male and female control rats and from rats treated with a single dose of recombinant human growth hormone (rhGH). No significant sex differences in the expression pattern were observed in both studied tissues, but STAT5 mRNA levels were significantly (P< 0.05) higher in liver than in thymus lymphocytes. STAT5 mRNA levels were significantly (P< 0.05) increased by a pulse of GH given to either male or female normal rats, suggesting a regulation of STAT5 gene expression in the studied tissues. In conclusion, quantitative solution hybridization-RNase protection assay of STAT5 mRNA provides a tool to further advance the study of the regulatory mechanisms involved in STAT5 gene expression. PMID- 11042020 TI - Echocardiographic assessment of cardiac morphology and function in mutant dwarf rats. AB - Although the mutant dwarf rat has been proposed as a model of growth hormone (GH) deficiency, few studies have addressed its cardiovascular abnormalities. Therefore, the aim of the present study was to investigate cardiac structure and function in mutant dwarf rats in vivo before and after chronic GH administration, by means of transthoracic Doppler echocardiography. To this purpose, forty 90-day old female dwarf rats were randomized to receive either GH treatment or placebo. Twenty age-and sex-matched Lewis rats (200-250 g) served as the control group. All rats underwent echocardiograms before receiving any drug and after 3 weeks of therapy. Echocardiographically detected left ventricular mass indexed to tibial length was reduced by 41% in dwarf rats compared to the control group. Such relative cardiac atrophy was also evident at the myocyte level, and was fully reversible after GH therapy. In contrast to the control group, dwarf rats also showed a reduction of left ventricular diastolic volumes normalized to tibial length and impaired cardiac performance as suggested by the reduction of cardiac index, abnormal stress-shortening relations, and a significant elevation of total peripheral vascular resistance. All these abnormalities were reversible upon GH therapy for 3 weeks. In conclusion, GH plays an important role in maintaining a normal cardiac structure and function. Since the observed changes are similar to those seen in GH-deficient men, the mutant dwarf rat represents a faithful animal model of GH deficiency. PMID- 11042021 TI - A novel bioassay based on human growth hormone (hGH) receptor mediated cell proliferation: measurement of 20K-hGH and its modified forms. AB - Previously we introduced the full-length hGH receptor (hGHR) into the mouse pro-B cell line, Ba/F3, and obtained stable transfectant (Ba/F3-hGHR), which could grow in response to 20K- and 22K-hGH in a dose-dependent manner(1). In the present study, we established a new bioassay system based on the proliferation of the Ba/F3-hGHR in combination with the eluted stain assay (ESTA). The Ba/F3-hGHR assay is completed in 18 h and requires only 10(-6)-fold amount of GH sample (1.8 ng) as compared with the rat weight gain assay. The validation study shows that the Ba/F3-hGHR assay is specific for hGH, precise (RSD = 1.1-19.7%) and ultrasensitive (lower limit of working range = 18.7 pg/mL). Four modified forms of recombinant 20K-hGH (oxidized, deamidated, des-Phe(1)and cleaved form) all of which are newly identified were measured by the Ba/F3-hGHR assay and the rat weight gain assay with our in-house recombinant 20K-hGH as standard. The oxidized and deamidated 20K-hGH were fully active, however the des-Phe(1)and cleaved 20K hGH had significantly reduced activities in both assays. These findings suggest that the Ba/F3-hGHR assay is useful as an alternative to the rat weight gain assay. PMID- 11042022 TI - Akt mediates insulin rescue from apoptosis in brown adipocytes: effect of ceramide. AB - We have recently shown that insulin can rescue serum deprived adipocytes from apoptosis in a PI 3 kinase and MAP kinase dependent manner. This study investigated the contribution of Akt and p70S6-kinase in insulin rescue from two different apoptotic triggers, serum deprivation and ceramide treatment. Insulin rescued serum-deprived immortalized brown adipocytes from apoptosis through phosphatidylinositol (PI) 3-kinase and Akt pathways, but independently of p70S6 kinase, as demonstrated by the use of inhibitors such as LY294002 or Rapamycin, and transfection experiments with dominant-negative constructs of Akt or p85 subunit of PI 3-kinase. A constitutively active Akt construct mimicked the insulin survival effect, decreasing the percentage of hypodiploid cells, the percentage of apoptopic cells and precluding the formation of apoptotic nuclei. We propose that the insulin survival effect on immortalized brown adipocytes is mediated through activation of Akt. However, insulin and EGF failed to rescue brown adipocytes from ceramide-induced apoptosis, as determined by DNA laddering, hypodiploid cells and apoptotic nuclei. Ceramide treatment blunted Akt activity but not PI 3-kinase activity, and insulin and EGF were unable to activate Akt. Ceramide also caused apoptosis in cells transfected with a constitutively active Akt construct, since phosphorylation of Akt was impaired under these experimental conditions. This study suggests that activation of Akt may be an absolute requirement for the survival of brown adipocytes. PMID- 11042023 TI - Impaired brain development and hydrocephalus in a line of transgenic mice with liver-specific expression of human insulin-like growth factor binding protein-1. AB - Insulin-like growth factors (IGFs) produced in the brain are known to participate in brain development via activation of the type 1 IGF receptor. IGF binding proteins (IGFBPs) modulate the cellular action of IGFs and some are expressed in the fetal brain. Under normal conditions IGFBP-1 is not one of these, but IGFBP-1 expression obtained via transgenesis using ubiquitous promoters affects brain development. In earlier work, we established a model of transgenic mouse in which liver-specific IGFBP-1 expression begins during fetal life. The repercussions of this IGFBP-1 over-expression include reproductive defects, ante- and perinatal mortality and post-natal growth retardation, the extent of which is related to the degree of transgene expression. Unexpectedly, during the first 2 months of postnatal life, there were some cases of head enlargement revealing hydrocephalus among homozygotes, frequently associated with motor disorders. Brain sections showed dilatation of the lateral ventricles in 10 out of 15 homozygotes examined. Histologically, dilatation was evident in four out of nine heterozygotes. Brain weight in transgenics was relatively less reduced than the weights of other organs. Hence, brain weight/body weight ratios were normal in heterozygotes and on average higher than normal in homozygotes. The width of the cerebral cortex was reduced in homozygotes, with disorganized neuronal layers. The corpus callosum was underdeveloped, particularly in homozygotes. The area of the hippocampus was reduced in homozygotes and one-third of the heterozygotes, with a short and thick dentate gyrus in the former. Similar anomalies have been reported in mice with disruption of the igf-I gene and in a model of transgenic mice over expressing IGFBP-1 in all tissues, including the brain. Hydrocephalus was not mentioned in these reports, raising the possibility that insertional mutagenesis may have been involved in our mice. Nevertheless, our observations indicate that hepatic over-expression of IGFBP-1 may have endocrine effects on brain development. PMID- 11042024 TI - Evidence for two distinct classes of high affinity growth hormone binding proteins in pregnant rat serum. AB - These studies have established the presence of two major classes of high affinity growth hormone binding proteins in pregnant rat serum, designated GHBPa and GHBPb, with apparent native Mr of 257 K and 98 K respectively. GHBPa, which has not been identified previously, exhibits a binding affinity (2-5 nM(-1)) that is up to 20-fold higher than GHBPb (0.2-0.8 nM(-1)) and is the least abundant form, being approximately 15-20% of total serum GH-binding capacity. Western immunoblot analysis revealed that each GHBP is composed of several immunoreactive proteins which were reactive with carboxy-terminal (RB1615) and/or N-terminal (MAb263) domain antibodies, suggesting the presence of GHBPs with and without the hydrophilic tail. Of importance is that GHBPa exhibited significantly higher Mr (78-182 K, +DTT) than that predicted by GHBP cloning, suggesting that they may be covalently bound to other non-GH-binding proteins or may be distinct entities. GHBPb, on the other hand, was composed of smaller Mr (43/48 K, +DTT) "hydrophilic" tail-containing proteins, some of which were disulphide linked to a larger complex of approximately 110 K. These novel findings challenge the current view of the mechanism for generation of the rat serum GHBP and raise the intriguing possibility that the two classes of GHBP may play distinct and important roles in GH physiology. PMID- 11042025 TI - Growth hormone deficiency in one of two siblings with Fanconi's anaemia complementation group FA-D. AB - Fanconi's anaemia (FA) shows great variability in phenotypic symptoms. We report on two FA siblings of German ancestry with the very rare form of the complementation group FA-D. Both presented with a similar phenotype and mild disease severity but with different growth. In the sister, growth velocity was normal, puberty and menarche occurred spontaneously. Her final height was within her parental target height. The younger brother had a reduced growth velocity, height SDS values below -5.5 SDS, a markedly retarded bone age, and delayed puberty. At the age of 12.9 years, growth hormone deficiency (GHD) was diagnosed and treatment with growth hormone was initiated. Our cases emphasize the heterogeneity of symptoms in FA even in siblings with the same genotype. In FA children with severe growth retardation, GHD must also be considered. PMID- 11042026 TI - Ligand blotting: iodinated vs biotinylated IGF. PMID- 11042027 TI - MIP-1alpha and TGF-beta production in CD34+ progenitor-stromal cell coculture systems: effects of progenitor isolation method and cell-cell contact. AB - ABSTRACT Macrophage inflammatory protein-1alpha (MIP-1alpha) is a C-C chemokine which has antiproliferative effects on early hematopoietic progenitors and stimulatory effects on later progenitors. It also possesses chemotactic and activating properties for monocytes, macrophages, and T-cells. CD34+ progenitors isolated utilizing an avidin-biotin immunoadsorption column produced significant amounts of MIP-1alpha from 24 h onward when cultured in medium with 10% fetal calf serum (>200 pg/ml). Such production persisted through 96 h of culture and was greater when such progenitors were cocultured with a preformed marrow stromal layer (4000 pg/ml at 24 h). The production of MIP-1alpha declined over time of coculture with stromal layers, and stromal layers themselves produced minimal MIP 1alpha as detected by ELISA: <100 pg/ml. In contrast, CD34+ cells isolated by flow cytometry or by magnetic bead adsorption produced minimal MIP-1alpha (0-30 pg/ml). MIP-1alpha production also increased when cells isolated by these two methods were cocultured with stromal layers. The difference in MIP-1alpha production could not be accounted for by differences in purity of the CD34+ population between isolation methods nor on the basis of monocytic or lymphocytic contamination as assessed by the presence of CD14 or CD3 positive cells. CD34+ cells isolated by immune adsorption had increased expression of endothelial and mesenchymal associated antigens, however, suggesting that this subpopulation might account for the MIP-1alpha production observed. Freshly isolated CD34+ cells expressed MIP-1alpha message as assessed by RT-PCR and by in situ hybridization. Coculture of CD34+ cells isolated by any means with stromal cells increased transforming growth factor-beta (TGF-beta) production, in this case by the stromal layer itself. Both MIP-1alpha and TGF-beta have been found to influence cell cycle status and proliferation status of early hematopoietic progenitors, and both have potential effects on accessory cell function. These studies indicate that progenitor-stromal cell interactions may influence local cytokine output, thus potentially influencing progenitor cycling status and accessory cell activation. The method of isolation of CD34+ progenitors may influence secretion of certain cytokines and chemokines. PMID- 11042028 TI - Distinct phenotypic expression associated with a new hyperunstable alpha globin variant (Hb heraklion, alpha1cd37(C2)Pro>0): comparison to other alpha thalassemic hemoglobinopathies. AB - Clinical phenotypes associated with abnormal globin chain biosynthesis may result in thalassemia (deficient quantity) or hemolytic anemia (abnormal hemoglobins). However, the phenotypic expression of hyperunstable hemoglobin variants often includes features of thalassemia, along with variable peripheral hemolysis. Hemoglobinopathies caused by highly unstable beta-chain variants have a dominant thalassemia-like phenotype, in which carriers have a clinical expression of thalassemia intermedia, but highly unstable alpha-globin variants are usually only phenotypically apparent when they interact with other alpha-thalassemia mutations. In a child with clinical and hematological features consistent with beta-thalassemia intermedia, DNA analysis excluded any beta-globin gene mutations but characterized a novel deletion cd37(C2)Pro>0 (Hb Heraklion) in the alpha1 globin gene, in trans to a common Mediterranean nondeletion alpha-thalassemia mutation (alpha(Hph)alpha). The deletion of proline at alpha37(C2) is predicted to result in severe instability of the variant hemoglobin, which on interaction with a synthesis-deficient alpha-thalassemia mutation causes a relatively severe dyserythropoietic anemia, representing an alternative phenotype associated with highly unstable alpha-chain variants. Hb Heraklion is the fourth highly unstable alpha-globin variant that we have observed in patients from Greece and Albania. Two variants involve the alpha2-globin gene: Hb Agrinio (alpha29(B10)Leu>Pro) and Hb Adana (alpha59(E8)Gly>Asp), and two the alpha1-gene: Hb Aghia Sophia (alpha62(E11)Val>0) and (Hb Heraklion a37(C2)Pro>0). Each has been observed on interaction with a different alpha-thalassemia mutation and the phenotypes associated with these highly unstable alpha-variants are presented. PMID- 11042029 TI - Comparative efficacy of dose regimens in enzyme replacement therapy of type I Gaucher disease. AB - Gaucher disease is caused by a deficiency of beta-glucocerebrosidase activity. The optimum dose and frequency of enzyme replacement therapy for Gaucher patients have not been determined. We set to compare the therapeutic effects of initiating treatment with macrophage-targeted glucocerebrosidase at a high dose followed by progressive dose reductions with that produced by initial treatment at a low dose in patients with type I Gaucher disease. The study included two parts: (i) Twelve patients received every 2 weeks enzyme replacement therapy at 60 IU/kg body wt for 24 months followed by sequential dose reduction every 6 months to 30 and then to 15 IU/kg body wt. (ii) Thirty-two patients received enzyme replacement therapy at 10 IU/kg every 2 weeks for 12 months. Hematologic parameters and liver and spleen volume were monitored in all patients. All patients had intact spleens. In patients who were started on high-dose enzyme replacement therapy, hemoglobin, acid phosphatase, and organ volume improved or remained unchanged at the end of each dose reduction. Platelet count decreased significantly when the dose of enzyme was reduced from 30 to 15 IU/kg body wt. Initiation of therapy at a low dose led to a significant improvement in all measured parameters at the end of 1 year. We conclude that the minimal effective dose for the nonskeletal manifestations of Gaucher disease can be achieved either by initiating enzyme replacement therapy with a high dose followed by a stepwise dose reduction or by starting treatment at the minimal dose. High dose provides a faster clinical response and should be considered for patients with more aggressive disease. The therapeutic threshold for macrophage-targeted glucocerebrosidase appears to be 10 15 IU/kg body wt every 2 weeks. PMID- 11042030 TI - Tissue- and epitope-specific mechanisms account for the diverse effects of anti CD44 antibodies on the maintenance of primitive hematopoietic progenitors in vitro. AB - The identification of rare stromal cells that support high levels of stem cells has opened avenues to identify molecules that contribute to the maintenance of these cells. We show that the maintenance of long-term culture initiating cells (LTC-IC) in stromal cell-supported cultures can be modulated via mAbs specific for CD44. mAb IM7.8.1 suppressed while mAb RAMBM44 enhanced LTC-IC levels in culture. Genetic polymorphisms in CD44 were used to show that the stromal cell compartment is targeted by mAb RAMBM44 and the hematopoietic compartment by mAb IM7.8. Neither of the CD44-specific mAbs inhibited adhesion of LTC-IC to the stroma, suggesting alternative mechanisms of action. In support of this interpretation, we show that mAb RAMBM44 directly induces signal transduction in the stromal cell line S17 but not in hematopoietic cells. Conversely, mAb IM7.8 elicited the appearance of phosphorylated bands in hematopoietic cells, but not in stromal cells. Collectively, the data indicate that the opposing effects of CD44-mediated regulation can be explained by different cellular programs that are elicited in distinct cell compartments. The binding of the enhancing mAb RAMBM44 to CD44 is specifically inhibited by collagen IV, while binding of the suppressive mAb IM7.8.1 is inhibited by a substance contained in the supernatant of the stromal cell line AC3.U. Thus, the CD44 epitopes defined by the mAbs bind distinct ligands and the ligands provide a potential physiological counterpart for the regulatory actions of the mAbs. PMID- 11042031 TI - Dosage-response in the treatment of Gaucher disease by enzyme replacement therapy. PMID- 11042032 TI - Identification of a novel recombinant allele in three unrelated Italian Gaucher patients: implications for prognosis and genetic counseling. AB - Gaucher disease (GD) results from deleterious mutations in the glucocerebrosidase gene. The relatively high frequency of some of these, especially at cDNA nucleotide 1226G (N370S) and at cDNA nucleotide 1448C (L444P), has led to the development of rapid screening techniques that can sometimes be misleading. In this report, we describe a novel rearrangement between the glucocerebrosidase gene and its pseudogene, identified as a consequence of a discrepancy between the genotype, homozygous for the common 1226G mutation, of an Italian patient with type 1 Gaucher disease, and the absence of the 1226G allele in her daughter. Additional investigations went on to reveal a novel recombinant allele beginning in intron 6 and extending through the rest of the coding sequence. Italian GD patients found homozygous for a specific mutation or with one or both alleles still unknown were further investigated and the novel recombinant allele was identified in an adult type 1 patient previously genotyped 1226G/1226G and in a young patient with an unknown genotype. The detection of this allele in three unrelated GD patients originating from the same geographic area in central Italy suggested a founder effect. This study emphasizes the implications of an accurate genotyping for the prognostic value of glucocerebrosidase genotype and reliable genetic counseling. PMID- 11042033 TI - Nramp2 analysis in hemochromatosis probands. AB - The mechanism that leads to iron overload in hereditary hemochromatosis is not yet fully understood and genes other than HFE may be involved. Nramp2 is an intestinal iron transporter, upregulated by dietary iron deficiency, which also colocalizes with transferrin in recycling endosomes. The purpose of the present study was to analyze the coding region of the Nramp2 gene in 14 hemochromatosis probands which did not carry any HFE mutations on both chromosomes. We confirmed the existence of a polymorphism (1254 T --> C), which presumably is not associated with hereditary hemochromatosis, but we did not find any mutation. On the other hand, we identified 17 splice variants of the Nramp2 mRNA. Eight corresponded to activation of cryptic splicing sequences between exons 3 and 4. They were observed in a majority of hemochromatosis probands and control subjects. This indicates the existence of an important splicing instability in this region. At this stage, the biological significance of these variants is unclear. Our study did not find evidence for the involvement of the Nramp2 gene in hereditary hemochromatosis. The remaining question is whether hemochromatosis probands in our study have iron overload because of environmental factors or due to mutation in gene(s) other than HFE and Nramp2. PMID- 11042034 TI - Delineation and mapping of Stat5 isoforms activated by granulocyte colony stimulating factor in myeloid cells. AB - Granulocyte colony-stimulating factor (G-CSF) is a cytokine critical for proliferation and differentiation of granulocytic precursors and neutrophil functions that has previously been demonstrated to activate Stat3 and Stat5, two members of the signal transducer and activator of transcription (STAT) protein family. Stat3 has been identified to be critical for G-CSF receptor (G-CSFR) mediated signaling for granulocyte differentiation. Stat5 activation has been mapped to the proximal portion of the cytosolic region of the G-CSFR. However, delineation and mapping of the specific Stat5 isoforms activated by G-CSF in myeloid cells have not been reported. In this study, we demonstrated that G-CSF activated a Stat5 complex in human myeloid cells containing three isoforms of Stat5: Stat5A, Stat5B, and Stat5 p80. Activation of Stat5A and Stat5B maps to the proliferation-specific domain of the G-CSFR, whereas Stat5 p80 is recruited by phosphotyrosine-704 within the region of G-CSFR required for differentiation. G CSF-activated Stat5A/B, but not Stat5 p80, formed a heterodimer with Stat3. The Stat5A/B-Stat3 heterodimer can bind to specific DNA sequences preferred by both Stat3 and Stat5. These findings are consistent with the possibility that Stat5 p80 contributes to G-CSF-induced myeloid differentiation. PMID- 11042035 TI - Hemoglobin C in transgenic mice: effect of HbC expression from founders to full mouse globin knockouts. AB - When present in the homozygous form, hemoglobin C (HbC, CC disease) increases red cell density, a feature that is the major factor underlying the pathology in patients with SC disease (Fabry et al., JCI 70, 1315, 1982). The basis for the increased red cell density has not yet been fully defined. We have generated a HbC mouse in which the most successful founder expresses 56% human alpha and 34% human beta(C). We introduced knockouts (KO) of mouse alpha- and beta-globins in various combinations. In contrast to many KO mice, all partial KOs have normal MCH. Full KOs that express exclusively HbC and no mouse globins have minimally reduced MCH (13. 7 +/- 0.3 pg/cell vs 14.5 +/- 1.0 for C57BL/6) and a ratio of beta- to alpha-globin chains of 0.88 determined by chain synthesis; hence, these mice are not thalassemic. Mice with beta(C) > 30% have increased MCHC, dense reticulocytes, and increased K:Cl cotransport. Red cell morphology studied by SEM is strikingly similar to that of human CC cells with bizarre folded cells. We conclude that red cells of these mice have many properties that closely parallel the pathology of human disease in which HbC is the major determinant of pathogenesis. These studies also establish the existence of the interactions with other gene products that are necessary for pleiotropic effects (red cell dehydration, elevated K:Cl cotransport, morphological changes) that are also present in these transgenic mice, validating their usefulness in the analysis of pathophysiological events induced by HbC in red cells. PMID- 11042036 TI - Linkage to Gaucher mutations in the Ashkenazi population: effect of drift on decay of linkage disequilibrium and evidence for heterozygote selection. AB - The two most common Gaucher disease mutations in the Ashkenazi population, 1226A- >G and 84G-->GG in the glucocerebrosidase gene, are tightly linked to a marker in the nearby pyruvate kinase gene. This paper develops a simulation of the Ashkenazi population that considers the effects of selection and drift on the mutant allele frequency and the recombinant haplotype frequency over time. Although the fraction of mutants that are linked to the original marker decays exponentially on average, this expected value is not very likely to occur. Instead, due to random loss of the recombinant haplotype, a mutation has a significant probability of retaining complete linkage disequilibrium long after its origin, so there may be large errors in estimating the age of a mutation based on linkage data. The simulations show that the 1226G mutation probably originated between 40 and 1000 generations ago (1000 to 25,000 years ago), and the 84GG mutation probably originated between 50 and 4800 generations ago (1300 to 120,000 years ago). The recent origin of the 1226G mutation and its high current allele frequency provide strong evidence for heterozygote selection. New techniques and results developed in this paper have general applicability toward analyzing linkage disequilibrium near other mutations. For example, they potentially explain the unexpected pattern of linkage disequilibrium seen around the DeltaF508 mutation of the cystic fibrosis transmembrane conductance regulator gene. PMID- 11042037 TI - Erythroid differentiation in vitro is blocked by cyclopamine, an inhibitor of hedgehog signaling. AB - Adult hematopoietic differentiation is a developmental process that employs many of the same molecular mechanisms as embryogenesis. To explore the possibility that hedgehog signaling is involved in the control of hematopoietic differentiation, we screened a panel of human leukemia cell lines for the expression of Patched1 and Smoothened, the receptor and coreceptor for hedgehog ligands. Expression was found in multiple cell lines, and Patched1 expression was detected in normal marrow. Induction of myeloid differentiation in cell lines downregulated expression of both genes. When normal marrow mononuclear cells were grown in semisolid medium in the presence of 10 microM cyclopamine, development of colonies of granulocytic/monocytic lineage was unaffected in terms of both number and morphology. The number of erythroid colonies, however, was significantly reduced (P < 0.01). Furthermore, hemoglobinization was substantially delayed relative to controls in those erythroid colonies that did form. Incubation of hematopoietic progenitors with Shh-N and GM-CSF resulted in increased granulocyte/monocyte colonies (P < 0.01); the increase was blocked by cyclopamine. Incubation of hematopoietic progenitors with Shh-N and stem cell factor resulted in larger erythroid colonies. These results suggest that elements of the hedgehog signaling pathway are involved in the control of hematopoietic differentiation. PMID- 11042038 TI - A study on the interaction between hydroxylamine analogues and oxyhemoglobin in intact erythrocytes. AB - The oxidative potency of hydroxylamine (HYAM) and its O-derivatives (O-methyl- and O-ethyl hydroxylamine) is generally larger than the effects of the N derivatives (N-methyl-, N-dimethyl-, and N,O-dimethyl hydroxylamine). The effects of the two groups of hydroxylamines also differ in a qualitative sense. To elucidate this difference in toxicity profiles we investigated the hemoglobin dependence of the toxicity, the occurrence of cell-damaging products like superoxide and H(2)O(2), and the cellular kinetics of the hydroxylamine analogues. All hydroxylamines were found to depend on the presence and accessibility of oxyhemoglobin to exert their toxicity. This did not provide an explanation for the different toxicity profiles. The interaction of some hydroxylamines with oxyhemoglobin is known to lead to the formation of radical intermediates. Differences in the stability of these radical products are known to occur, and in some cases secondary products are formed. This can contribute to the differences in toxicity. In this respect, production of superoxide radicals was demonstrated for all hydroxylamines in the reaction with oxyhemoglobin. Evidence for H(2)O(2) generation during the reaction of HYAM, O-methyl, O-ethyl-, and N-dimethyl hydroxylamine with oxyhemoglobin was also found. Next to variations in the products formed, differences in cellular kinetics are likely to be among the most important factors that explain the different toxicity patterns seen for the hydroxylamines in erythrocytes. Indeed, differences were found to exist for the kinetics of methemoglobin formation in erythrocytes. Not only was the final level of methemoglobin formed much lower for the N-derivatives, but also the reaction rate with oxyhemoglobin was slower than with HYAM and its O derivatives. Except for N,O-dimethyl hydroxylamine (NODMH), the same pattern was seen in hemolysates. NODMH tripled its effect on hemoglobin in hemolysate compared with incubations in erythrocytes. This implies that cellular uptake is a limiting factor for NODMH. Since formation of H(2)O(2) is most likely a result of an interaction with hemoglobin, differences in kinetics of methemoglobin formation can be an explanation for the fact that NMH and NODMH did not produce H(2)O(2) to a detectable level. These results indicate that (a) the toxicity of all hydroxylamines depends on an interaction with oxyhemoglobin; (b) the interaction with hemoglobin produces radical intermediates and concomitantly superoxide radicals and H(2)O(2); and (c) differences in uptake, reaction rate with hemoglobin, and stability of the intermediates formed do exist for the different hydroxylamines and contribute to their differences in toxicity. PMID- 11042039 TI - Glucose-6-phosphate dehydrogenase mutations and haplotypes in Mexican Mestizos. AB - In a screening for glucose-6-phosphate dehydrogenase (G-6-PD) deficiency in 1985 unrelated male subjects from the general population (Groups A and B) belonging to four states of the Pacific coast, 21 G-6-PD-deficient subjects were detected. Screening for mutations at the G-6-PD gene by PCR-restriction enzyme in these 21 G-6-PD-deficient subjects as well as in 14 G-6-PD-deficient patients with hemolytic anemia belonging to several states of Mexico showed two common G-6-PD variants: G-6-PD A-(202A/376G) (19 cases) and G-6-PD A-(376G/968C) (9 cases). In 7 individuals the mutations responsible for the enzyme deficiency remain to be determined. Furthermore, four silent polymorphic sites at the G-6-PD gene (PvuII, PstI, 1311, and NlaIII) were investigated in the 28 individuals with G-6-PD A- variants and in 137 G-6-PD normal subjects. As expected, only 10 different haplotypes were observed. To date, in our project aiming to determine the molecular basis of G-6-PD deficiency in Mexico, 60 unrelated G-6-PD-deficient Mexican males-25 in previous studies and 35 in the present work-have been studied. More than 75% of these individuals are from states of the Pacific coast (Sinaloa, Nayarit, Jalisco, Michoacan, Guerrero, Oaxaca, and Chiapas). The results show that although G-6-PD deficiency is heterogeneous at the DNA level in Mexico, only three polymorphic variants have been observed: G-6-PD A-(202A/376G) (36 cases), G-6-PD A-(376G/968C) (13 cases), and G-6-PD Seattle(844C) (2 cases). G-6-PD A- variants are relatively distributed homogeneously and both variants explain 82% of the overall prevalence of G-6-PD deficiency. The variant G-6-PD A (202A/376G) represents 73% of the G-6-PD A- alleles. Our data also show that the variant G-6-PD A-(376G/968C)-which has been observed in Mexico in the context of two different haplotypes-is more common than previously supposed. The three polymorphic variants that we observed in Mexico are on the same haplotypes as found in subjects from Africa, the Canary Islands, and Spain. PMID- 11042040 TI - The effects of RARalpha and RXRalpha proteins on growth, viability, and differentiation of v-myb-transformed monoblasts. AB - Retinoids are important agents which regulate differentiation and proliferation processes in various cell types, including cancer cells. Growth arrest and induction of terminal differentiation demonstrate the tumor-suppressive effects of retinoids on leukemic cells. We studied differentiation, proliferation, and death processes in the cell line of v-myb-transformed monoblasts BM2 and their retinoic acid receptor (RAR) alpha- and retinoid X receptor (RXR) alpha expressing derivatives after exposure to four different retinoids: all-trans retinoic acid, 9-cis retinoic acid, TTNPB, and LG1000153. The effects of retinoids on the phenotype of BM2, BM2RAR, and BM2RXR cells were correlated with the transcription activation function of the v-Myb oncoprotein of avian myeloblastosis virus. We found that the efficiency of terminal differentiation of BM2RAR and BM2RXR cells induced by retinoids is indirectly proportional to the v Myb transcription activation activity. In contrast, the effects of liganded retinoid receptors on growth of BM2 cells are more complex. Activated RAR protein induces growth inhibition of BM2 cells by suppression of v-Myb function. However, liganded RXR protein is less efficient in cell cycle arrest and rather decreases cellular viability. This process can occur in the presence of active v-Myb protein. These results suggest that ligand-activated RARalpha protein is primarily engaged in control of proliferation and differentiation of v-myb transformed monoblasts, while activated RXRalpha protein controls their differentiation and death. PMID- 11042041 TI - Protein hydration and location of water molecules in oxidized horse heart cytochrome c by (1)H NMR. AB - The hydration properties of the oxidized form of horse heart cytochrome c have been studied by (1)H NMR spectroscopy. Two-dimensional, homonuclear ePHOGSY-NOESY experiments are used to map water-protein interactions. The detected NOEs reveal interactions between nonexchangeable protein protons and both water protons and labile protein protons which exchange with water protons. Among the many water molecules apparent in the X-ray structure, three have been identified with a residence time longer than 300 ps. One of them is located inside the distal heme cavity, in the deepest part of a hydration pathway extending toward the surface. The identification of hydrophilic regions and detection of three long-lived water molecules settles some ambiguities and provides a better representation of the water-protein interactions in oxidized cytochrome c. PMID- 11042042 TI - Atomic refinement using orientational restraints from solid-state NMR. AB - We describe a procedure for using orientational restraints from solid-state NMR in the atomic refinement of molecular structures. Minimization of an energy function can be performed through either (or both) least-squares minimization or molecular dynamics employing simulated annealing. The energy, or penalty, function consists of terms penalizing deviation from "ideal" parameters such as covalent bond lengths and terms penalizing deviation from orientational data. Thus, the refinement strives to produce a good fit to orientational data while maintaining good stereochemistry. The software is in the form of a module for the popular refinement package CNS and is several orders of magnitude faster than previous software for refinement with orientational data. The short computer time required for refinement removes one of the difficulties in protein structure determination with solid-state NMR. PMID- 11042043 TI - Gradient-echo line scan imaging using 2D-selective RF excitation. AB - A gradient-echo line scan imaging technique was developed which employs two dimensional spatially selective radiofrequency (2DRF) pulses for consecutively exciting individual columns of transverse magnetization, i.e., image lines. Although a variety of trajectories are possible for 2DRF excitation, the current implementation involved a blipped-planar trajectory in conjunction with additional saturation RF pulses to suppress side excitations above and below the desired image section, i.e., along the blip direction of the 2DRF pulse. Human brain imaging at 2.0 T (Siemens Vision, Erlangen, Germany) resulted in measuring times of 5.2 s for a 5-mm section at 1.0 x 1.0 mm in-plane resolution. Functional neuroimaging of the motor cortex at 1.2 s temporal resolution and 0.78 x 1.56 mm in-plane resolution exploited the capability of imaging inner volumes (here a 25 mm strip) without signal aliasing. PMID- 11042044 TI - Measurement of internuclear distances in solid-state NMR by a background-filtered REDOR experiment AB - A background-filtered version of the rotational-echo double resonance (REDOR) experiment is demonstrated. The experiment combines a traditional REDOR pulse sequence with a double-cross-polarization (DCP) sequence to select only those signals coming from spin pairs of interest. The relatively inefficient DCP sequence, which transfers polarization from (1)H to (15)N and subsequently to (13)C, is improved by the use of adiabatic passages through the (-1) sideband of the Hartmann-Hahn matching condition. The result is an efficient 2D-REDOR pulse sequence that does not require a reference experiment for removal of background signals. The data produced by the experiment are ideally suited to analysis by newly developed dipolar transform methods, such as the REDOR transform. The relevant features of the experiment are demonstrated on simple labeled amino acids. Relative efficiencies of several other potential filtering methods are also compared. Copyright 2000 Academic Press. PMID- 11042045 TI - Thermally polarized (1)H NMR microimaging studies of liquid and gas flow in monolithic catalysts AB - The feasibility of gas flow imaging in moderately high magnetic fields employing thermally polarized gases at atmospheric pressures is demonstrated experimentally. Two-dimensional spatial maps of flow velocity distributions for acetylene, propane, and butane flowing along the transport channels of shaped monolithic alumina catalysts were obtained at 7 T by (1)H NMR, with true in-plane resolution of 400 &mgr;m and reasonable detection times. The resolution is shown to be limited by the echo attenuation due to rapid molecular diffusion in the imaging gradients of magnetic field. All gas flow images exhibit flow patterns that are not fully developed, in agreement with the range of Reynolds numbers (190-570) and the length of the sample used in gas flow experiments. The flow maps reveal the highly nonuniform spatial distribution of shear rates within the monolith channels of square cross-section, the kind of information essential for evaluation and improvement of the efficiency of mass transfer in shaped catalysts. The water flow images were obtained at lower Re numbers for comparison. These images demonstrate the transformation of a transient flow pattern observed closer to the inflow edge of a monolith into a fully developed one further downstream. Copyright 2000 Academic Press. PMID- 11042046 TI - Effects of various types of molecular dynamics on 1D and 2D (2)H NMR studied by random walk simulations AB - By carrying out random walk simulations we systematically study the effects of various types of complex molecular dynamics on (2)H NMR experiments in solids. More precisely, we calculate one-dimensional (1D) (2)H NMR spectra and the results of two dimensional (2D) (2)H NMR experiments in time domain, taking into account isotropic as well as highly restricted motions which involve rotational jumps about different finite angles. Although the dynamical models are chosen to mimic the primary and secondary relaxation in supercooled liquids and glasses, we do not intend to describe experimental results quantitatively but rather to show general effects appearing for complex reorientations. We carefully investigate whether 2D (2)H NMR in time domain, which was originally designed to measure correlation times of ultraslow motions (tau >/= 1 ms), can be used to obtain shorter tau, too. It is demonstrated that an extension of the time window to tau >/= 10 &mgr;s is possible when dealing with exponential relaxation, but that it will fail if there is a distribution of correlation times G(lgtau). Vice versa, we show that 1D (2)H NMR spectra, usually recorded to look at dynamics with tau in the microsecond regime, are also applicable for studying ultraslow motions provided that the loss of correlation is achieved step by step. Therefore, it is useful to carry out 1D and 2D NMR experiments simultaneously in order to reveal the mechanism of complex molecular motions. In addition, we demonstrate that highly restricted dynamics can be clearly observed in 1D spectra and in 2D NMR in time domain if long solid-echo delays and large evolution times are applied, respectively. Finally, unexpected observations are described which appear in the latter experiment when considering very broad distributions G(lgtau). Because of these effects, time scale and geometry of a considered motion cannot be extracted from a straightforward analysis of experimental results. Copyright 2000 Academic Press. PMID- 11042047 TI - Simulation studies of high-field EPR spectra of spin-labeled lipids in membranes. AB - The high-field (i.e., 94 GHz) membrane EPR spectra of lipids spin labeled in their fatty acid chains have been simulated by using two limiting motional models. The aim was to identify the dynamic origin of the residual (g(xx) - g(yy)) anisotropy observed in the nonaxial EPR spectra of cholesterol-containing membranes. It is concluded that the residual spectral anisotropy arises from in plane ordering of the lipid chains by cholesterol. The partial averaging of the (g(xx) - g(yy)) anisotropy was best described by restricted axial rotation with a frequency in the region of tau(-1)(R||) approximately 0.5-1 x 10(9) s(-1). Simulations for slower axial rotation of unrestricted amplitude produced less satisfactory fits. In phospholipid membranes not containing cholesterol, the nonaxial anisotropy is completely averaged in the fluid phase and substantially reduced even in the gel phase. The unrestricted axial rotation in the gel phase is of comparable frequency to that of the limited axial rotation in the liquid ordered phase of membranes containing cholesterol. These results on in-plane ordering by cholesterol in the liquid-ordered phase could be significant for current proposals regarding domain formation in cellular membranes. PMID- 11042048 TI - MAS double-quantum filtered dipolar shift correlation spectroscopy AB - A carbon-13 magic angle spinning double-quantum filtered dipolar shift correlation NMR experiment which can be used to establish through-space connectivities in solids is analyzed. The main advantage of the double-quantum filtered approach is the removal of intensity arising from natural abundance background signals. The variation in intensity of the cross and diagonal peaks observed in the two-dimensional spectrum as a function of mixing time is investigated experimentally for model systems. The observed behavior is compared with analytical expressions derived for three coupled spins, as well as with simulations based on average Hamiltonian theory. Copyright 2000 Academic Press. PMID- 11042049 TI - Multiple-time correlation functions in spin-3/2 solid-state NMR spectroscopy AB - Stimulated echo spectroscopy of nonselectively excitable I = 3/2 nuclei offers new perspectives for the investigation of ultraslow motions predominantly in inorganic solids and solid-like materials. Conditions for the generation of pure, quadrupole modulated multipolar spin orders and for the detection of two- and four-time correlation functions are discussed. The case of spins I > 3/2 is also briefly considered. Copyright 2000 Academic Press. PMID- 11042050 TI - Relationships between (1)H NMR relaxation data and some technological parameters of meat: a chemometric approach. AB - In this paper chemometrics (ANOVA and PCR) is used to measure unbiased correlations between NMR spin-echo decays of pork M. Longissimus dorsi obtained through Carr-Purcell-Meiboom-Gill (CPMG) experiments at low frequency (20 MHz) and the values of 14 technological parameters commonly used to assess pork meat quality. On the basis of the ANOVA results, it is also found that the CPMG decays of meat cannot be best interpreted with a "discrete" model (i.e., by expanding the decays in a series of a discrete number of exponential components, each with a different transverse relaxation time), but rather with a "continuous" model, by which a continuous distribution of T(2)'s is allowed. The latter model also agrees with literature histological results. PMID- 11042051 TI - Relaxation of nuclear magnetization in a nonuniform magnetic field gradient and in a restricted geometry AB - We study the influence of restriction on Carr-Purcell-Meiboom-Gill spin echo response of magnetization of spins diffusing in a bounded region in the presence of a nonuniform magnetic field gradient. We consider two fields in detail-a parabolic field which, like the uniform-gradient field, scales with the system size, and a cosine field which remains bounded. Corresponding to three main length scales, the pore size, L(S), the dephasing length, L(G), and the diffusion length during half-echo time, L(D), we identify three main regimes of decay of the total magnetization: motionally averaged, localization, and short-time. In the short-time regime (L(D) << L(S), L(G)), we confirm that the leading order behavior is controlled by the average of the square of the gradient, (nablaB(z))(2), and in the motionally averaged regime (MAv), where L(S) << L(D), L(G), by (integral dxB(z))(2). We verify numerically that two different fields for which those two averages are identical result in very similar decay profiles not only in the limits of short and long times but also in the intermediate times, with important practical implications. In the motionally averaged regime we found that previous estimates of the decay exponent for the parabolic field, based on a soft-boundary condition, are significantly altered in the presence of a more realistic, hard wall. We find the scaling of the decay exponent in the MAv regime with pore size to be L(2)(S) for the cosine field and L(6)(S) for the parabolic field, as contrasted with the linear gradient scaling of L(4)(S). In the localization regime, for both the cosine and the parabolic fields, the decay exponent depends on a fractional power of the gradient, implying a breakdown of the second cumulant or the Gaussian phase approximation. We also examined the validity of time-evolving the total magnetization according to a distribution of effective local gradients and found that such approximation works well only in the short-time regime and breaks down strongly for long times. Copyright 2000 Academic Press. PMID- 11042052 TI - Dipolar interactions in molecules aligned by strong AC electric fields AB - We observed magnetization transfer and spectroscopic splittings due to dipolar couplings in the solution NMR spectra of neat nitrobenzene aligned using AC electric fields. Weak dipolar splittings have been previously observed for nitrobenzene in a DC electric field (T. M. Plantenga, et al., Chem. Phys. 66, 1 9, 1982); the use of homogeneous pulsed AC fields has allowed us to establish stable experimental conditions, which were more tolerable to sample impurities and required no sample purification, and to carry out multidimensional experiments. A pulse sequence is discussed in which the electric field is present only for the indirect dimension: this sequence records the dipolar splittings for each proton in the indirect dimension; the direct dimension presents the isotropic chemical shift. Another pulse sequence is discussed that uses the applied electric field only in the mixing period to produce cross peaks between dipolar coupled pairs and correlate their isotropic chemical shifts. The order parameter describing molecular alignment was in good agreement with that previously determined from deuterium quadrupolar measurements of deuterated nitrobenzene in a similar range of electric fields: S(mol) approximately 0.025% for a field strength of 7.0 MV/m (rms). The dipolar splittings for ortho-meta, meta-para, and ortho-para protons were in qualitative agreement with the known geometry. Copyright 2000 Academic Press. PMID- 11042053 TI - Synchronized adiabatic decoupling. AB - A new decoupling scheme termed "synchronized adiabatic decoupling" is developed for use in the indirectly detected dimension. After each increment, the decoupling sequence is replaced by another one with different period T or different initial period T(ini) so that sampling always occurs at the end of a complete decoupling period. The effects of J coupling are therefore completely averaged out for all data points. As a result, all decoupling sidebands disappear and the center band increases correspondingly. Since the synchronized adiabatic decoupling does not require conventional editing techniques to cancel the sidebands, it is useful in high-field gradient-enhanced multidimensional experiments with only a single scan per increment. PMID- 11042055 TI - Analysis and prediction of isotropic mixing magnetization transfer profiles in three-spin topologies AB - Isotropic mixing transfer functions (T(kl)) in three-spin systems typical of amino acids have been analyzed in order to develop simple rules for predicting transfer maxima/minima. For certain topologies, the intrinsically complex expressions describing the transfer functions reduce to compact forms which are easy to interpret and analyze. For other topologies where compactification is not possible, an analysis of the component functions of the T(kl) reveals that only one or two components contribute significantly to the overall profile of the transfer function. As a result, simple rules of the thumb may be devised for reasonably accurate prediction of mixing times corresponding to local and global transfer maxima/minima, thereby facilitating mixing time optimization in TOCSY experiments. Copyright 2000 Academic Press. PMID- 11042054 TI - Rapid acquisition transverse relaxometric imaging. AB - Segmented echo-planar acquisitions have been incorporated into a multiecho imaging sequence to produce a MRI method for rapid transverse relaxometry. The method is demonstrated on gel phantoms and rat brain and found to produce unbiased estimates of T(2). Gradient performance can be a limiting factor for the implementation of this technique and there is a cost in signal-to-noise ratio resulting from the higher bandwidth required, as is typical for echo-planar acquisitions. PMID- 11042056 TI - RRT: the regularized resolvent transform for high-resolution spectral estimation AB - A new numerical expression, called the regularized resolvent transform (RRT), is presented. RRT is a direct transformation of the truncated time-domain data into a frequency-domain spectrum and is suitable for high-resolution spectral estimation of multidimensional time signals. One of its forms, under the condition that the signal consists only of a finite sum of damped sinusoids, turns out to be equivalent to the exact infinite time discrete Fourier transformation. RRT naturally emerges from the filter diagonalization method, although no diagonalization is required. In RRT the spectrum at each frequency s is expressed in terms of the resolvent R(s)(-1) of a small data matrix R(s), that is constructed from the time signal. Generally, R is singular, which requires certain regularization. In particular, the Tikhonov regularization, R(-1) approximately [R(dagger)R + q(2)](-1)R(dagger) with regularization parameter q, appears to be computationally both efficient and very stable. Numerical implementation of RRT is very inexpensive because even for extremely large data sets the matrices involved are small. RRT is demonstrated using model 1D and experimental 2D NMR signals. Copyright 2000 Academic Press. PMID- 11042057 TI - HYDRONMR: prediction of NMR relaxation of globular proteins from atomic-level structures and hydrodynamic calculations. AB - The heteronuclear NMR relaxation of globular proteins depends on the anisotropic rotational diffusion tensor. Using our previous developments for prediction of hydrodynamic properties of arbitrarily shaped particles, by means of bead models, we have constructed a computational procedure to calculate the rotational diffusion tensor and other properties of proteins from their detailed, atomic level structure. From the atomic coordinates file used to build the bead model, the orientation of the pertinent dipoles can be extracted and combined with the hydrodynamic information to predict, for each residue in the protein, the relaxation times. All of these developments have been implemented in a computer program, HYDRONMR, which will be of public domain. PMID- 11042058 TI - Coherence transfer signals in the rotational resonance NMR of a spinning single crystal AB - A recent analysis of rotational resonance lineshapes (M. Helmle et al., J. Magn. Reson. 140, 379-403, 1999) predicted the existence of coherence transfer signals, which are generated by mechanically induced coherence transfer during the detection process. These signals correspond to the generation of observable coherences at spin sites that have no magnetization at the beginning of the observation interval but which acquire coherence while the detection is underway. The coherence transfer signals disappear for powder samples in conventional magic angle-spinning solid-state NMR experiments. In this Communication, we report the successful detection of coherence transfer signals in rotor-synchronized experiments performed on a single crystal of [1,2-(13)C(2)]glycine. Copyright 2000 Academic Press. PMID- 11042059 TI - Message from the editors PMID- 11042060 TI - Preface PMID- 11042061 TI - Introduction. Where neuroscience has been and where it needs to go. PMID- 11042062 TI - Mapping brain terrain. PMID- 11042063 TI - Deception of perception. PMID- 11042064 TI - Pain. PMID- 11042065 TI - Hearing and deafness. PMID- 11042066 TI - The plastic brain. PMID- 11042067 TI - Simple organisms. PMID- 11042068 TI - Spinocerebellar ataxias. PMID- 11042069 TI - Genes, gene expression, and behavior. PMID- 11042070 TI - Expressive genes record memories. PMID- 11042071 TI - Basic science in psychiatry. PMID- 11042072 TI - Stress hormones: good and bad. PMID- 11042073 TI - Drug addiction. PMID- 11042074 TI - Alzheimer's disease. PMID- 11042075 TI - Epilepsy. PMID- 11042076 TI - Stroke. PMID- 11042077 TI - Parkinson's disease. PMID- 11042078 TI - Two different oligomeric states of the RuvB branch migration motor protein as revealed by electron microscopy. AB - In prokaryotes, the RuvA, B, and C proteins play major roles at the late stage of DNA homologous recombination, where RuvB complexed with RuvA acts as an ATP dependent motor for branch migration. The oligomeric structures of negatively stained and frozen hydrated RuvB from Thermus thermophilus HB8 were investigated by electron microscopy. RuvB oligomers free of DNA formed a ring structure of about 14 nm in diameter. The averaged top view image clearly indicated a sevenfold symmetry, suggesting that it exists as a heptamer. The RuvB oligomers complexed with duplex DNA formed a smaller ring of about 13 nm in diameter. The averaged top view images represented a sixfold symmetry. This difference in oligomerization indicates that the oligomeric structure of RuvB may convert from a heptamer to a hexamer upon DNA binding. In addition, this finding provides the lesson that great care should be taken in investigating the subunit organizations of DNA binding proteins, because their oligomeric states are more sensitive to DNA interactions than expected. PMID- 11042079 TI - Structural characterization of penicillin-binding protein-related factor A (PrfA) from Bacillus species. AB - The prfA genes of Bacillus stearothermophilus and Bacillus subtilis are in an operon downstream of the ponA gene encoding penicillin-binding protein 1 (PBP1), a major enzyme involved in peptidoglycan synthesis. The specific function of the 23- to 24-kDa PrfA protein is unknown but this protein plays some role in nucleoid segregation and the functions of PrfA and PBP1 are interrelated. We overexpressed B. stearothermophilus and B. subtilis PrfA in Escherichia coli and purified the proteins to homogeneity by cation exchange and gel filtration chromatography. The protein is a monomer in solution, and circular dichroism spectroscopy revealed an abundance of beta-sheet secondary structure. Crystals of B. stearothermophilus PrfA were also obtained and diffracted X-rays to 1.8 A resolution. PMID- 11042080 TI - Expression, two-dimensional crystallization, and three-dimensional reconstruction of the beta8 outer membrane protein Omp21 from Comamonas acidovorans. AB - The Omp21 protein from the proteobacterium Comamonas (Delftia) acidovorans belongs to the recently described beta8 family of outer membrane proteins, characterized by eight antiparallel beta-strands which form a beta-barrel. This family includes virulence proteins, OmpA and OmpX from Escherichia coli, and other related molecules. After we established an expression system, recombinant Omp21 was purified by Ni(2+) chelation affinity chromatography and refolded in situ while bound to resin. The native state of refolded protein was proven by FTIR spectroscopy and monitored with denaturing PAGE (heat modification). Both native and recombinant Omp21 were reconstituted in lipid membranes and crystallized two-dimensionally by controlled dialysis. Recombinant Omp21 crystallized as dimer and formed a p22(1)2(1) lattice with constants of a = 11.1 nm, b = 12.2 nm, gamma = 89.5 degrees. The 3-D structure of negatively stained, recombinant Omp21 was determined at a resolution of 1.8 nm by means of electron crystallography. Comparison with 3-D maps of OmpX and the transmembrane domain of OmpA revealed a high similarity between the mass distribution of exoplasmic loops of Omp21 and OmpA. PMID- 11042081 TI - Chimeric human-simian anti-CD4 antibodies form crystalline high symmetry particles. AB - A chimeric human-simian IgG, antigen specific for CD4, when exposed to 0.5 M SO(=)(4) containing 0.4% polyethylene glycol or Jeffamine, self-assembles into discreet, roughly spherical particles 23 nm in diameter. Increasing SO(=)(4) to 1.55 M induces the IgG particles to crystallize in either a hexagonal or a monoclinic form. From X-ray diffraction, the former crystal is of space group P622, with one IgG particle in the unit cell; thus the particle itself must have 622 point group symmetry. Both crystal forms have been imaged using atomic force microscopy. Detailed features of the duodecamer were evident, including the symmetry and a large solvent channel along the sixfold axis. The particles in some ways resemble the hexameric IgG aggregates believed to activate compliment upon antigen binding and, therefore, may have physiological relevance. Investigation of seven other IgGs of diverse origins and subclasses indicates that many, if not most, IgGs form similar particles. To our knowledge, this is the first observation of the assembly of IgG into high symmetry aggregates in the absence of antigen or their crystallization. PMID- 11042082 TI - Ultrastructure of sea urchin calcified tissues after high-pressure freezing and freeze substitution. AB - The improvements brought by high-pressure freezing/freeze substitution fixation methods to the ultrastructural preservation of echinoderm mineralized tissues are investigated in developing pedicellariae and teeth of the echinoid Paracentrotus lividus. Three freeze substitution (FS) protocols were tested: one in the presence of osmium tetroxide, one in the presence of uranyl acetate, and the last in the presence of gallic acid. FS in the presence of osmium tetroxide significantly improved cell ultrastructure preservation and should especially be used for ultrastructural studies involving vesicles and the Golgi apparatus. With all protocols, multivesicular bodies, suggested to contain Ca(2+), were evident for the first time in skeleton-forming cells. FS in the presence of gallic acid allowed us to confirm the structured and insoluble character of a part of the organic matrix of mineralization in the calcification sites of the tooth, an observation which modifies the current understanding of biomineralization control in echinoderms. PMID- 11042083 TI - Local average intensity-based method for identifying spherical particles in electron micrographs. AB - A method is presented that reliably detects spherical viruses from a wide variety of noisy low-contrast electron micrographs. Such detection is one of the first image analysis steps in the computer-aided reconstruction of three-dimensional density distribution models of viruses. Particle detection is based on the comparison of intensity in a circular area and in the surrounding ring followed by a number of tests to validate the potential particles. The only required input from the user in addition to the micrograph is an approximate radius of the particle. The method has been implemented as program ETHAN that has been tested for several different data sets. ETHAN has also successfully been used to detect DNA-less virus particles for an actual reconstruction. PMID- 11042084 TI - Human and Xenopus cingulin share a modular organization of the coiled-coil rod domain: predictions for intra- and intermolecular assembly. AB - The complete nucleotide and derived amino acid sequences of Homo sapiens cingulin cDNA (5143 bp) were determined by sequencing two distinct EST clones that showed significant sequence homology to Xenopus laevis cingulin. Protein sequence analysis indicates that the molecule contains two chains and has a tripartite structure with N-terminal (head) domains, a coiled-coil rod domain (length, 120 nm), and short C-terminal (tail) domains. Human and Xenopus cingulin heads are only 33% identical, yet a human cingulin N-terminal fragment still interacts with canine ZO-1 and ZO-2 in vitro. The rod domain contains two A and two B subdomains, though it lacks the third B subdomain present in Xenopus cingulin. The heptad substructures of Xenopus and human cingulins were further characterized by computer analysis and indicated that the two-stranded coiled coil structure contained chains that were parallel and in axial register. Fast Fourier transform analysis and a scoring technique designed to recognize potential interactions between different supramolecular arrangements suggests that cingulin dimers may further assemble through antiparallel interactions between the last approximately 100 amino acids of the coiled-coil region. Cingulin mRNA ( approximately 5.2 kb) was detected by Northern blotting in epithelial tissues. A human cingulin EST was mapped to chromosome 1q21 using the UniGene database. PMID- 11042085 TI - Structural analysis of the bacteriophage T3 head-to-tail connector. AB - The connector protein of bacteriophage T3, p8, has been overexpressed in Escherichia coli. Purification of the oligomers built by several copies of p8 reveals a mixed population of dodecamers and tridecamers. The percentages of these two types of oligomers differ in every culture growth, indicating that assembly of this protein depends upon the conditions of the expression system. Those cultures that generated a majority of dodecamers allowed, after purification of the connectors, the two-dimensional crystallization of the dodecamers in a tetragonal arrangement, while the tridecamers did not form crystals. The processing and averaging of several images of frozen-hydrated crystals and their internal phase comparison shows that the crystals are arranged in a P42(1)2 space group, with cell unit dimensions of 165 x 165 A. The three dimensional reconstruction generated with images of crystals ranging from 0 degrees to 60 degrees tilt reveals a wide domain surrounded by 12 protrusions and a narrow domain that serves to interact with the tail of the bacteriophage. A channel runs along the connector wide enough to allow the translocation of a double-stranded DNA molecule into the prohead. The general structure of the T3 connector is very similar to those obtained for other nonrelated bacteriophages and strongly suggests that the shape of this important viral structure is intimately related to its function. PMID- 11042086 TI - Preliminary crystallographic study of a complex formed between the alpha/beta tubulin heterodimer and the neuronal growth-associated protein SCG10. AB - Crystals of a complex formed between the alpha/beta-tubulin heterodimer and SCG10, a neuron-specific growth-associated protein, have been obtained by the hanging drop method. They belong to the space group P2(1)2(1)2(1), with unit cell parameters a = 56 A, b = 353 A, c = 466 A and four molecular complexes in the asymmetric unit. A complete X-ray diffraction data set to 6.1 A resolution has been collected using synchrotron radiation. This represents a challenging opportunity to study at a molecular level the structure-function relationships between a microtubule-destabilizing protein, SCG10, and tubulin. PMID- 11042087 TI - Crystallization and preliminary X-ray analysis of receptor-binding protein P2 of bacteriophage PRD1. AB - Bacteriophage PRD1 has remarkable structural similarities to adenovirus, but is unusual in containing a membrane beneath its icosahedral capsid. Its monomeric receptor-binding protein, P2, is part of a complex at each capsid vertex and so is the functional equivalent of adenovirus fiber. P2 has been crystallized by the "hanging-drop" method of vapor diffusion and two different crystal forms were obtained. Macroseeding, used to increase the size of the initial small needles, gave rod-shaped crystals. These grew to a size of 0.08 x 0.08 x 0.50 mm(3) and diffracted to 2.6 A resolution. They have the orthorhombic space group P222(1), with unit cell dimensions a = 137.8 A, b = 46.5 A, c = 136.4 A. A few single crystals of a second form were grown without seeding under slightly different conditions. A parallelepiped crystal (0.10 x 0.10 x 0.35 mm(3)), with space group C222(1) and unit cell dimensions a = 182.3 A, b = 204.8 A, c = 133.3 A, diffracted to 3.5 A resolution. A rotation function for the second form revealed that four monomers of P2 are related by a noncrystallographic twofold axis. The structure of P2 will reveal how this arrangement relates to the trimeric adenovirus fiber. PMID- 11042088 TI - Preliminary X-ray crystallographic and NMR studies on the exonuclease domain of the epsilon subunit of Escherichia coli DNA polymerase III. AB - The structured core of the N-terminal 3'-5' exonuclease domain of epsilon, the proofreading subunit of Escherichia coli DNA polymerase III, was defined by multidimensional NMR experiments with uniformly (15)N-labeled protein: it comprises residues between Ile-4 and Gln-181. A 185-residue fragment, termed epsilon(1-185), was crystallized by the hanging drop vapor diffusion method in the presence of thymidine-5'-monophosphate, a product inhibitor, and Mn(2+) at pH 5.8. The crystals are tetragonal, with typical dimensions 0.2 mm x 0.2 mm x 1.0 mm, grow over about 2 weeks at 4 degrees C, and diffract X-rays to 2.0 A. The space group was determined to be P4(n)2(1)2 (n = 0, 1, 2, 3), with unit cell dimensions a = 60.8 A, c = 111.4 A. PMID- 11042089 TI - Effect of arsenite on the induction of CYP1A4 and CYP1A5 in cultured chick embryo hepatocytes. AB - We had reported previously that 2.5-5 microM sodium arsenite decreased the phenobarbital-mediated induction of CYP2H activity and protein but not CYP2H1 mRNA in chick-embryo hepatocyte cultures. Induction of a CYP1A activity and protein by 3-methylcholanthrene was also decreased by low arsenite concentrations; however, CYP1A mRNAs were not measured in those studies. We report here that low concentrations of arsenite decreased induction of activities and mRNAs of two chicken cytochromes P450, CYP1A (1A4 and 1A5), by 3 methylcholanthrene in chick-embryo hepatocyte cultures. Arsenite treatment did not affect the turnover of either mRNA, nor did it decrease the superinduction of each mRNA caused by treatment with cycloheximide in addition to 3 methylcholanthrene. Glutathione depletion enhanced the effect of arsenite to decrease induction of CYP1A4. These results indicate the induction of CYP1A4 and 1A5 is inhibited by sodium arsenite at the level of transcription, suggesting that the Ah receptor complex may be involved. PMID- 11042090 TI - A plasticizer released from IV drip chambers elevates calcium levels in neurosecretory terminals. AB - We report that intracellular calcium levels rise in mammalian neurosecretory terminals and in cultured pheochromocytoma cells during acute exposure to physiological medium incubated in IV drip chambers. The agent responsible for this effect is shown to be di(2-ethylhexyl)phthalate (DEHP). DEHP (800 nM) added to saline solution caused a rise in [Ca(2+)](i) similar to that elicited by the contaminant-containing solution. The extraction of this contaminant from the IV drip chamber, as measured by spectrophotometry, was time-dependent and was markedly accelerated by the presence of 50 mM ethanol in the solution. Larger [Ca(2+)](i) increases were observed in terminals exposed to solutions incubated in IV drip chambers for greater durations. The rise in calcium requires transmembrane calcium flux through membrane channels, as the response is blocked by either 100 microM cadmium or by lowering the extracellular free Ca(2+) concentration to 10 microM. Our results suggest that acute alterations in intracellular calcium should be considered in addition to long-term effects when determining the safety of phthalate-containing plastics and that laboratory researchers using plastic perfusion materials consider this potential source of artifactual results. PMID- 11042091 TI - Human skin absorption and metabolism of the contact allergens, cinnamic aldehyde, and cinnamic alcohol. AB - trans-Cinnamaldehyde and trans-cinnamic alcohol have been commonly reported to cause allergic contact dermatitis (ACD) in humans. Cinnamaldehyde is a more potent skin sensitizer than cinnamic alcohol. It has been hypothesized that cinnamic alcohol is a "prohapten" that requires metabolic activation, presumably by oxidoreductase enzymes such as alcohol dehydrogenase (ADH) or cytochrome P450 2E1 (CYP2E1), to the protein-reactive cinnamaldehyde (a hapten). In this study, the in vitro percutaneous absorption and metabolism of cinnamaldehyde and cinnamic alcohol (78 micromol dose) has been examined using freshly excised, metabolically viable, full-thickness breast and abdomen skin from six female donors. Penetration rates and total cumulative recoveries of cinnamic compounds that were present in receptor fluid, extracted from within the skin, evaporated from the skin surface, or remained unabsorbed on the skin surface after 24 h were quantified by reversed-phase high-performance liquid chromatography. Biotransformation of cinnamaldehyde to both cinnamic alcohol and cinnamic acid was observed. Topically applied cinnamic alcohol was converted to cinnamaldehyde (found on the skin surface only) and cinnamic acid. To establish whether these biotransformations were enzymatic, experiments were performed in the absence and presence of varying concentrations (80-320 micromol) of the ADH/CYP2E1 inhibitors pyrazole or 4-methylpyrazole. The observation that pyrazole significantly reduced (p < 0.05) the total penetration of cinnamic metabolites into receptor fluid, following either cinnamaldehyde or cinnamic alcohol treatment, but did not significantly affect parent chemical penetration, suggests that we are measuring cutaneous metabolic products of ADH activity. The skin absorption and metabolism of cinnamaldehyde and cinnamic alcohol will play an important role in the manifestation of ACD following topical exposure to these compounds. PMID- 11042092 TI - Activation of mitogen-activated protein kinases by tributyltin in CCRF-CEM cells: role of intracellular Ca(2+). AB - Effects of tributyltin chloride (TBT) and other organotin compounds on mitogen activated protein kinases (MAPKs) were examined in CCRF-CEM human T lymphoblastoid cells. In response to the incubation with 0.25-2 microM TBT for 1 h, the levels of the phosphorylated form of extracellular signal-regulated protein kinase (ERK), c-Jun NH(2)-terminal kinase (JNK), and p38 MAPK increased in a dose-dependent manner. The phosphorylation was observed after 15 min and lasted for 4 h following exposure to 1 microM TBT, while the cell viability was not lowered significantly within 6 h. On the other hand, no clear changes were found in the total protein levels of ERK, JNK, and p38 MAPK. The in vitro activities of MAPKs also increased in response to TBT exposure. The potentials of MAPKs phosphorylation and of cellular damage were TBT > dibutyltin dichloride (DBT) > monobutyltin trichloride (MBT). When compared to other triorganotin compounds such as trimethyltin chloride (TMT), triphenyltin chloride (TPT), and triethyltin bromide (TET), TBT exposure induced the most marked phosphorylation of MAPKs. Chelation of intracellular Ca(2+) suppressed TBT-induced MAPKs phosphorylation almost completely, but removal of external Ca(2+) did not. The present results showed that tributyltin is a potent activator of ERK, JNK, and p38 MAPK pathways, and Ca(2+) mobilized from intracellular stores plays an important role for the phosphorylation of MAPKs in this human T cell line. PMID- 11042093 TI - Pulmonary effects induced by ultrafine PTFE particles. AB - PTFE (polytetrafluoroethylene) fumes consisting of large numbers of ultrafine (uf) particles and low concentrations of gas-phase compounds can cause severe acute lung injury. Our studies were designed to test three hypotheses: (i) uf PTFE fume particles are causally involved in the induction of acute lung injury, (ii) uf PTFE elicit greater pulmonary effects than larger sized PTFE accumulation mode particles, and (iii) preexposure to the uf PTFE fume particles will induce tolerance. We used uf Teflon (PTFE) fumes (count median particle size approximately 16 nm) generated by heating PTFE in a tube furnace to 486 degrees C to evaluate principles of ultrafine particle toxicity. Teflon fumes at ultrafine particle concentrations of 50 microg/m(3) were extremely toxic to rats when inhaled for only 15 min. We found that when generated in argon, the ultrafine Teflon particles alone are not toxic at these exposure conditions; neither were Teflon fume gas-phase constituents when generated in air. Only the combination of both phases when generated in air caused high toxicity, suggesting either the existence of radicals on the surface or a carrier mechanism of the ultrafine particles for adsorbed gas compounds. Aging of the fresh Teflon fumes for 3.5 min led to a predicted coagulation to >100 nm particles which no longer caused toxicity in exposed animals. This result is consistent with a greater toxicity of ultrafine particles compared to accumulation mode particles, although changes in particle surface chemistry during the aging process may have contributed to the diminished toxicity. Furthermore, the pulmonary toxicity of the ultrafine Teflon fumes could be prevented by adapting the animals with short 5-min exposures on 3 days prior to a 15-min exposure. Messages encoding antioxidants and chemokines were increased substantially in nonadapted animals, yet were unaltered in adapted animals. This study shows the importance of preexposure history for the susceptibility to acute ultrafine particle effects. PMID- 11042094 TI - Ethnic differences in human flavin-containing monooxygenase 2 (FMO2) polymorphisms: detection of expressed protein in African-Americans. AB - The flavin-containing monooxygenases (FMOs) are a family of xenobiotic metabolizing enzymes that are expressed in a species- and tissue-specific manner. FMO2 expression has been observed in pulmonary tissue from several species, but not human. Two human FMO2 point mutations have been reported: a cytosine to thymidine transition at position 1414 resulting in a premature stop codon and a thymidine insertion at position 1589 resulting in a frameshift. To define the frequency of these sequence variations and explore their significance, unrelated African-American, Caucasian, and Korean individuals were genotyped. In the African-American population tested (n = 180), the 1414C allele occurred at a 13% frequency; however, all of the tested Caucasians (n = 52) and Koreans (n = 100) were homozygous for the 1414T allele. The T1589 allele occurred at frequencies of 6.9 and 13.0% in African-Americans (n = 175) and Caucasians (n = 23), respectively, and appears to segregate with the 1414T allele. Thus, it would have no further impact on FMO2 activity. Western blot analysis of pulmonary microsomes failed to detect immunoreactive protein in 1414T homozygotes. A heterozygotic individual did exhibit a single band of the expected size, but no detectable FMO activity in the corresponding lung microsomes. Sequence analysis, however, was consistent with the 1414C allele encoding an active FMO2 enzyme. FMO2 mRNA expression was observed in most individuals, but failed to correlate with genotype or protein expression. In summary, functional FMO2 is expressed in only a small percentage of the overall population. However, in certain ethnic groups, active pulmonary FMO2 enzyme will be present in a significant number of individuals. PMID- 11042095 TI - Metabolism and disposition of bisphenol A in female rats. AB - Bisphenol A (BPA), which is used in the manufacture of polycarbonates, elicits weak estrogenic activity in in vitro and in vivo test systems. The objectives of this study were to compare the patterns of disposition of radioactivity in adult female F-344 and CD rats after oral administration of (14)C BPA (100 mg/kg), to isolate the glucuronide of BPA and to assess its estrogenic activity in vitro, and to evaluate the transfer of radioactivity to pups from lactating dams administered (14)C BPA. Over 6 days, F-344 rats excreted more radioactivity in urine than CD rats. The major metabolite in urine was identified as bisphenol A glucuronide (BPA gluc) by incubation with beta-glucuronidase and (1)H and (13)C NMR spectroscopy. In lactating CD rats administered (14)C BPA (100 mg/kg) by gavage, only a small fraction of the label was found in milk, with 0.95 +/- 0.66, 0.63 +/- 0.13, and 0.26 +/- 0.10 microg equiv/ml (mean +/- SD) from dams collected 1, 8, and 26 h after dosing, respectively. Radioactivity in pup carcasses indicated exposure in the range of microgram equivalents per kilogram; those values ranged from 44.3 +/- 24.4 for pups separated from their lactating dams at 2 h to 78.4 +/- 10.9 at 24 h. BPA gluc was the prominent metabolite in milk and plasma. In test systems for activation of in vitro estrogen receptors alpha and beta, BPA gluc did not show appreciable efficacy at concentrations up to 0.03 mM, indicating that metabolism via glucuronidation is a detoxication reaction. PMID- 11042096 TI - A pharmacokinetic model for predicting absorption, elimination, and tissue burden of toxaphene in rats. AB - A two-compartment pharmacokinetic model was formulated to predict absorption, elimination, and tissue burden of toxaphene in rats. The model was constructed based on the database of Crowder and Dindal (Bull. Environ. Contam. Toxicol. 12, 320-327, 1974) and included six tissue compartments: blood, brain, liver, muscle, fat, and carcass. The pharmacokinetically based dosimetry indicated that absorption of toxaphene was fast in fat, whole body, carcass, and blood, relatively slow in liver and muscle, and slow in brain. In contrast, the elimination rate was rapid in whole body, muscle, and blood, moderate in carcass and brain, and slow in liver and fat. Tissue burden was highest in fat, whole body, and blood, intermediate in liver, and lowest in brain. The model performance was evaluated by the data set of Pollock and Hillstrand (J. Environ. Sci. Health B 17, 635-648, 1982) on toxaphene absorption and elimination in pregnant rats. Validity of the model was confirmed by the close agreement between the predicted and observed tissue burdens of toxaphene in target tissues. Disposition of toxaphene via feces was a dominant excretory pathway while urinary excretion was a minor elimination route in male rats. However, for pregnant rats, excretion of toxaphene both in urine and feces were of similar magnitude. These characteristics of elimination are valuable for understanding the metabolism of toxaphene in pregnant rats. The model serves as a starting point for a quantitative, mechanism-based understanding of the processes that influence the pharmacokinetics of toxaphene in mammalian systems. PMID- 11042097 TI - Dietary genistein inactivates rat thyroid peroxidase in vivo without an apparent hypothyroid effect. AB - Biological effects of genistein are currently under investigation by the National Toxicology Program because of widespread and increasing soy consumption by humans and evidence for modulation of endocrine function. Rats were exposed to genistein aglycone in soy-free feed fortified at 0, 5, 100, and 500 ppm starting in utero through 20 weeks. Thyroid glands and serum were analyzed for total genistein (aglycone + conjugates) using HPLC with electrospray mass spectrometric detection. Microsomal thyroid peroxidase (TPO) activity was measured spectrophotometrically. The total genistein content in rat serum was as high as 8 microM, and significant dose-dependent increases of genistein in thyroid tissue up to 1 pmol/mg were found in male and female rats. The activity of TPO in male and female rats was found to be reduced by up to 80% in a dose-dependent manner. Male and female rats consuming a standard soy-based rodent diet (NIH 31) had TPO activity approximately 50% lower than rats consuming a soy-free diet and this loss was commensurate with measured serum levels of isoflavones. Suicide inactivation of rat, porcine, and human TPO was observed in vitro at concentrations of genistein aglycone comparable to those measured in rat thyroids. Thyroid hormone levels (T3, T4, TSH) in serum, thyroid weights, and histopathology showed no differences between treated and untreated groups. These findings suggest that, even though substantial amounts of TPO activity are lost concomitant to soy isoflavone consumption by normal rats, the remaining enzymatic activity is sufficient to maintain thyroid homeostasis in the absence of additional perturbations. PMID- 11042098 TI - Development of phase II xenobiotic metabolizing enzymes in differentiating murine clara cells. AB - Glutathione S-transferases (GSTs) and epoxide hydrolases (EHs) protect cells from exogenous insult by detoxifying electrophilic compounds. Little is known about these enzyme systems during postnatal lung development. This study was designed to help establish whether the heightened neonatal susceptibility of the lung to bioactivated cytotoxicants is the result of inadequate ability to detoxify reactive intermediates. We compared the distribution of immunoreactive protein and enzymatic activity of GSTs and EHs in isolated distal airways during pre- and postnatal development in lungs of mice from 16 days gestation to 9 weeks postnatal age (adult). GST alpha, mu, and pi class protein expression in fetal and postnatal lung varied by isozyme and age. Isozymes alpha and mu are expressed at low levels before birth, high levels on postnatal day 7, low levels between postnatal days 14 and 21, high levels at postnatal day 28, and slightly lower levels in adults. Immunoreactive protein of isozyme pi has a peak expression on gestational day 18 and again on postnatal day 4, is undetectable at postnatal day 21, and is at peak levels in the adult mouse lung. GST activity in distal airways increased with age. Microsomal EH protein expression increased in intensity with age, while activity was similar in airways from all ages. We conclude that in the mouse lung (1) cellular expression of glutathione S-transferase varies by age and isozyme and does not increase with increasing age, (2) airway glutathione S transferase activity increases with increasing age and does not correlate with immunoreactive protein expression, and (3) airway microsomal epoxide hydrolase activity does not increase, even though immunoreactive protein expression does increase with age. PMID- 11042100 TI - Chemical index for volume 168 PMID- 11042101 TI - Optimal intraguild foraging and population stability. AB - This article explores effects of adaptive intraguild predation on species coexistence and community structure in three species' food webs. Two Lotka Volterra models that assume a trade-off between competition and predation strength are considered in detail. The first model does not explicitly model resource dynamics and is considered with both nonadaptive and adaptive intraguild predation; in the latter case predators choose their diet in order to maximize their instantaneous population growth rate. The second model includes resource population dynamics. Effects of adaptive intraguild predation on the community structure along a gradient in environment productivity are analyzed and compared with some experimental results of protist food webs. Conditions under which intraguild predation is adaptive are discussed for both models. It is proved that if intraguild predators are perfect optimizers then intraguild predation should decrease with increasing environmental productivity and adaptive intraguild predation is a stabilizing factor provided environmental productivity is high enough. PMID- 11042099 TI - Analysis of the cytotoxic properties of linoleic acid metabolites produced by renal and hepatic P450s. AB - Cytochrome P450 epoxidation of linoleic acid produces biologically active metabolites which have been associated with many pathological conditions that often lead to acute renal failure. In the present study, we evaluated the ability of specific cytochrome P450s to produce linoleic acid monoepoxides. We then tested the cytotoxic properties of linoleic acid, linoleic acid monoepoxides, and corresponding diols in a rabbit renal proximal tubule model. CYP1A2, CYP2E1, CYP2J2, CYP2J3, CYP2J5, and CYP2J9 metabolized linoleic acid at rates comparable to arachidonic acid and produced linoleic acid monoepoxides as major products. Cytotoxicity studies showed that linoleic acid, linoleic acid monoepoxides, and corresponding diols are toxic at pathologically relevant concentrations (100-500 microM). Concentration-dependent studies showed that linoleic acid and linoleic acid monoepoxides are the most toxic and induce mitochondrial dysfunction prior to cell death. Cytoprotectants known to block cell death associated with mitochondrial dysfunction and oxidative stress did not prevent cell death induced by linoleic acid and linoleic acid monoepoxides. This study shows that P450s in the CYP1 and CYP2 gene families metabolize linoleic acid to linoleic acid monoepoxides and that the monoepoxides, as well as linoleic acid, disrupt mitochondrial function without causing oxidative stress. PMID- 11042102 TI - Periodic triggering of an inducible gene for control of a wild population. AB - A possible method of control for the management of wild populations consists of continual introgression of an inducible transgene by releasing transgenic individuals, with periodic exposure of the population to a trigger. Exposure to the trigger causes death or sterility in carriers of the transgene, but is otherwise benign. We investigate the effectiveness of various strategies for control. We show that suppression of the population density below any pre specified level is possible using this technique. At the same time we show that too frequent or too efficient exposure to the trigger can select for non transgenic genotypes at an intensity such that the population density will be largely unaffected by the trigger. Choices for management parameters can ensure that the latter scenario is avoided. We show that releasing individuals carrying the transgene at more than one locus facilitates density control. PMID- 11042103 TI - The cytonuclear effects of facultative apomixis. I. Disequilibrium dynamics in diploid populations. AB - We comprehensively analyze the cytonuclear effects of generalized mixed mating, including all combinations of selfing, outcrossing, and apomixis, the asexual production of seeds. After first deriving the time-dependent solutions for nonrandom associations (disequilibria) between a diallelic cytoplasmic marker and the alleles and genotypes at a diploid nuclear locus, we delimit all possible dynamical behaviors and the conditions under which each occurs. As in standard mixed mating systems, all disequilibria ultimately decay to zero except when outcrossing is absent, in which case permanent disequilibria result if the allelic association is initially nonzero. When at least some outcrossing is present, any initial allelic association decays at a constant geometric rate, whereas genotypic disequilibria may first increase in magnitude or change sign. Although selfing and apomixis tend to retard the decay of disequilibria (or approach to equilibrium) and often to the same extent, apomixis can have a stronger effect under some conditions. We also determine the dynamics of cytonuclear disequilibria in specific examples that may be of particular interest for empirical studies of hybrid zones. The results suggest several practical guidelines for experimental design and data analysis and show how the cytonuclear disequilibrium dynamics under mating system alone furnish a quantitative baseline for null hypotheses against which to test for the presence of other evolutionary forces. PMID- 11042104 TI - The cytonuclear effects of facultative apomixis. II. Definitions and dynamics of disequilibria in tetraploid populations. AB - We develop a cytonuclear framework for tetraploid populations in which a diallelic nuclear marker exhibits tetrasomic inheritance. This system requires two separate parameterizations, with six cytonuclear disequilibria (nonrandom associations) in tetraploid individuals and four in their diploid gametes. Double reduction during meiosis adds further complexity by causing gametic output to vary with the distance of the nuclear locus from the centromere. We derive and analyze dynamical solutions for the disequilibria under generalized mixed mating, with any combination of apomixis, selfing, and outcrossing, with and without double reduction. As in comparable diploid systems, all disequilibria ultimately decay to zero, unless nuclear and cytoplasmic alleles are nonrandomly associated and outcrossing is absent, in which case permanent associations result. Selfing and apomixis retard the decay of disequilibria (or approach to equilibrium), and often to the same extent. In contrast, double reduction can accelerate the loss of tetraploid cytonuclear associations, but only negligibly in hybrid zones, and this loss is never faster than in diploids. Only in the absence of allelic associations or outcrossing is the asymptotic approach to equilibrium differentially affected by apomixis and selfing or slower under tetrasomic than disomic inheritance. To facilitate empirical applications, we also examine tetraploid hybrid zone dynamics and offer practical guidelines for experimental design and data analysis, showing how the consequences of the mating system alone provide a valuable baseline for drawing evolutionary inferences from the observed patterns of cytonuclear associations. PMID- 11042105 TI - Kin selection and natal dispersal in an age-structured population. AB - We examine the effect of iteroparity on the evolution of dispersal for a species living in a stable but fragmented habitat. We use a kin selection model that incorporates the effects of demographic stochasticity on the local age structure and age-specific genetic identities. We consider two cases: when the juvenile dispersal rate is allowed to change with maternal age and when it is not. In the latter case, we find that the unconditional evolutionarily stable dispersal rate increases when the adult survival rate increases. Two antagonistic forces act upon the evolution of age-specific dispersal rates. First, when the local age structure varies between patches of habitat, the intensity of competition between adults and juveniles in the natal patch is, on average, lower for offspring born to older senescent mothers. This selects for decreasing dispersal with maternal age. Second, offspring born to older parents are on average more related to other juveniles in the same patch and they experience a higher intensity of kin competition, which selects for increasing dispersal with maternal age. We show that the evolutionary outcome results from a balance between these two opposing forces, which depends on the amount of variance in age structure among sub populations. PMID- 11042106 TI - Cultural transmission in a demographic study of sex ratio at birth in China's future. AB - A decline in fertility causes an increase in the sex ratio at birth (SRB) in countries with strong son preference. What happens to the SRB if fertility is maintained at a low level depends on the evolution of son preference. In this paper, we analyze trends in son preference and its effect on China's future SRB. PMID- 11042107 TI - Amino-acid-dependent signal transduction. AB - Recent research carried out in several laboratories has indicated that, in addition to their role as intermediates in many metabolic pathways, amino acids can interact with insulin-dependent signal transduction. In this short review, the current state of this rapidly expanding field is discussed. PMID- 11042108 TI - Multitasking in signal transduction by a promiscuous human Ins(3,4,5,6)P(4) 1 kinase/Ins(1,3,4)P(3) 5/6-kinase. AB - We describe a human cDNA encoding 1-kinase activity that inactivates Ins(3,4,5,6)P(4), an inhibitor of chloride-channel conductance that regulates epithelial salt and fluid secretion, as well as membrane excitability. Unexpectedly, we further discovered that this enzyme has alternative positional specificity (5/6-kinase activity) towards a different substrate, namely Ins(1,3,4)P(3). Kinetic data from a recombinant enzyme indicate that Ins(1,3,4)P(3) (K(m)=0.3 microM; V(max)=320 pmol/min per microg) and Ins(3,4,5,6)P(4) (K(m)=0.1 microM; V(max)=780 pmol/min per microg) actively compete for phosphorylation in vivo. This competition empowers the kinase with multitasking capability in several key aspects of inositol phosphate signalling. PMID- 11042109 TI - Identification of multiple proteins expressed in murine embryos as binding partners for the WW domains of the ubiquitin-protein ligase Nedd4. AB - Nedd4 is a member of a growing family of ubiquitin-protein ligases which consist of a lipid-binding domain, two to four WW domains and a C-terminal ubiquitin protein ligase domain. The Nedd4 mRNA levels are developmentally regulated and Nedd4 protein is highly expressed in many mouse embryonic tissues. In this study we have used a far-Western screen to identify embryonic proteins that interact with the WW domains in mouse Nedd4. We report here identification of eight Nedd4 WW-domain-interacting proteins from mouse embryonic cDNA expression libraries. Two of the proteins are novel, while two have been identified previously as ligands for a WW domain. All of these proteins contain one or more PY motifs. In seven of the eight proteins, these PY motifs are necessary for their interaction with the WW domains of Nedd4. Using site-directed mutagenesis, and by using individual WW domains of Nedd4 as probes for far-Western analysis, we show that the three WW domains in Nedd4 interact with varying affinities with the PY motifs present in various Nedd4-binding proteins. These results provide evidence that Nedd4 can potentially interact with multiple proteins, possibly simultaneously, through its WW domains. PMID- 11042110 TI - An examination of how structural changes can affect the rate of electron transfer in a mutated bacterial photoreaction centre. AB - A series of reaction centres bearing mutations at the (Phe) M197 position were constructed in the photosynthetic bacterium Rhodobacter sphaeroides. This residue is adjacent to the pair of bacteriochlorophyll molecules (P(L) and P(M)) that is the primary donor of electrons (P) in photosynthetic light-energy transduction. All of the mutations affected the optical and electrochemical properties of the P bacteriochlorophylls. A mutant reaction centre with the change Phe M197 to Arg (FM197R) was crystallized, and a structural model constructed at 2.3 A (1 A=0.1 nm) resolution. The mutation resulted in a change in the structure of the protein at the interface region between the P bacteriochlorophylls and the monomeric bacteriochlorophyll that is the first electron acceptor (B(L)). The new Arg residue at the M197 position undergoes a significant reorientation, creating a cavity at the interface region between P and B(L). The acetyl carbonyl substituent group of the P(M) bacteriochlorophyll undergoes an out-of-plane rotation, which decreases the edge-to-edge distance between the macrocycles of P(M) and B(L). In addition, two new buried water molecules partially filled the cavity that is created by the reorientation of the Arg residue. These waters are in a suitable position to connect the macrocycles of P and B(L) via three hydrogen bonds. Transient absorption measurements show that, despite an inferred decrease in the driving force for primary electron transfer in the FM197R reaction centre, there is little effect on the overall rate of the primary reaction in the bulk of the reaction-centre population. Examination of the X-ray crystal structure reveals a number of small changes in the structure of the reaction centre in the interface region between the P and B(L) bacteriochlorophylls that could account for this faster-than-predicted rate of primary electron transfer. PMID- 11042111 TI - Stichopus japonicus arginine kinase: gene structure and unique substrate recognition system. AB - Arginine kinase from the sea cucumber Stichopus japonicus underwent a unique molecular evolution. Unlike the monomeric 40 kDa arginine kinases from molluscs and arthropods, Stichopus arginine kinase is dimeric, the same as cytoplasmic isoenzymes of the vertebrate creatine kinases. Its entire amino acid sequence is more similar to creatine kinases than to other arginine kinases, but the guanidino specificity region (GS region) is of the arginine kinase type. To elucidate its unusual evolution, the structure of the Stichopus arginine kinase gene was determined. It consisted of seven exons and six introns, and a part of the exon 2 of the Stichopus gene corresponds to the GS region. Compared with the structure of the human muscle creatine kinase gene (seven exons, six introns), the splice junctions of five introns were conserved exactly between the two genes, suggesting that these introns had been conserved for at least 500 million years. The entire sequence of Stichopus arginine kinase is distinctly included in the creatine kinase cluster in all tree construction methods examined. On the other hand, if the tree is constructed only from sequences corresponding to Stichopus exon 2, it is placed in the arginine kinase cluster. Thus we conclude that Stichopus arginine kinase evolved not from the arginine kinase gene but from the creatine kinase gene, and suggest that its GS region, determining substrate specificity, has been replaced by an arginine kinase type via exon shuffling. In typical arginine kinases four residues, Ser(63), Gly(64), Val(65) and Tyr(68) (numbering from the Limulus polyphemus sequence), in the GS region are highly conserved and are associated with substrate binding. Among them, Tyr(68) appears to play a crucial role by forming a hydrogen bond with the substrate, and is conserved exactly in all arginine kinases. However, in Stichopus arginine kinase, none of these four conserved residues were present. Nevertheless, the enzyme displays an affinity for the substrate arginine (K(m)=0.8 mM) comparable with other arginine kinases. This implies that a completely different substrate binding system has been developed in Stichopus arginine kinase. We propose that the His(64) in Stichopus arginine kinase acts as a substitute for the Tyr(68) in other arginine kinases, and that the imidazole ring of His(64) is hydrogen bonded with the substrate arginine, thus stabilizing it. PMID- 11042112 TI - Aberrant phosphorylation of dentatorubral-pallidoluysian atrophy (DRPLA) protein complex in brain tissue. AB - Dentatorubral-pallidoluysian atrophy (DRPLA) is caused by DRPLA protein which carries expansion of a glutamine repeat. Abnormal high-molecular-mass complex formation by DRPLA protein and its pathological ubiquitination comprise the disease processes in the brains of patients with DRPLA. In this study, DRPLA protein complex was isolated and shown to have pathologically stronger bond formation with DRPLA proteins in DRPLA brain tissue compared with control brain tissue. Immunochemical methods and an enzymic dephosphorylation technique were used to demonstrate that DRPLA protein complex is aberrantly phosphorylated in DRPLA brain tissue. Immunohistochemical studies show that both the ubiquitinated cytoplasmic inclusions and the nuclear membrane are aberrantly phosphorylated in DRPLA-affected neurons. This finding suggests that the nuclear membrane is another pathological focus of DRPLA neurodegeneration. PMID- 11042113 TI - The pH dependence of naturally occurring low-spin forms of methaemoglobin and metmyoglobin: an EPR study. AB - The paramagnetic species in human metHb and horse metmyoglobin (metMb) have been studied at low temperature using EPR spectroscopy. The high-spin (HS) haem signal in aquometMb has a greater rhombic distortion than the HS metHb signal. Nevertheless, the individual line width (g=6) is smaller in metMb than in metHb, consistent with non-identical signals from the alpha and beta Hb subunits. Three low-spin (LS) haem forms are present in metHb, while metMb has only two. The major LS form in both proteins is the alkaline species (with OH(-) at the sixth co-ordination position). The minor LS forms are assigned to different histidine hemichromes in equilibrium with the normal HS species at low temperature. LS forms disappear when the haem is bound by a ligand, such as fluoride, which ensures 100% occupancy of the HS state both at room temperature and at 25 K. The small differences in effective g-factors of the histidine hemichromes are interpreted in terms of different distances between the distal histidine and haem iron. The pH dependence of the inter-conversion of the different paramagnetic species is consistent with a model whereby protonation of a residue with a pK of 5.69 (metHb) or 6.12 (metMb), affects ligand binding and transformation from the HS to the LS form. Chemical and spectroscopic considerations suggest that the residue is unlikely to be the proximal or distal histidine. We therefore propose a model where protonation of this distant amino acid causes a conformational change at the iron site. Identical effects are seen in frozen human blood, suggesting that this effect may have physiological significance. PMID- 11042114 TI - Transmembrane and cytoplasmic domains of syndecan mediate a multi-step endocytic pathway involving detergent-insoluble membrane rafts. AB - Syndecan heparan sulphate proteoglycans directly mediate a novel endocytic pathway. Using Chinese hamster ovary cells expressing the human syndecan 1 core protein or a chimaeric receptor, FcR-Synd, consisting of the ectodomain of the IgG Fc receptor Ia linked to the transmembrane and cytoplasmic domains of syndecan 1, we previously reported that efficient internalization is triggered by ligand clustering, requires intact actin microfilaments and tyrosine kinases, proceeds with a t(1/2) of approx. 1 h and is distinct from coated-pit pathways. We have now examined the involvement of cholesterol-rich, detergent-insoluble membrane rafts. On clustering, (125)I-labelled IgG bound to FcR-Synd rapidly became insoluble in cold Triton X-100, well before endocytosis. Insolubility of clustered FcR-Synd ligand did not require the syndecan ectodomain, linkage of the cytoplasmic tail to the cytoskeleton, or energy-dependent cellular metabolism. Pretreatment of cells with cyclodextrin to deplete cholesterol from rafts abolished insolubility of the clustered ligand and inhibited endocytosis in a dose-responsive fashion. Similar results were obtained with (125)I-labelled lipoprotein lipase bound to authentic cell-surface syndecan. In contrast, the 39 kDa receptor-associated protein (RAP), a coated-pit ligand, was more than 80% soluble in cold Triton even after internalization; cellular cholesterol depletion failed to substantially affect the internalization of (125)I-RAP. Overall, our results indicate a multi-step endocytic process consisting of ligand binding, clustering, energy-independent lateral movement into detergent-insoluble membrane rafts and finally recruitment of actin and tyrosine kinases to bring the ligands into the cell. PMID- 11042115 TI - Insulin-induced phospholipase D1 and phospholipase D2 activity in human embryonic kidney-293 cells mediated by the phospholipase C gamma and protein kinase C alpha signalling cascade. AB - Phospholipase D (PLD)1 is quiescent in vitro and in vivo until stimulated by classical protein kinase C (PKC) isoforms, ADP-ribosylation factor or Rho family members. By contrast, PLD2 has high basal activity, and the mechanisms involved in agonist-induced activation of PLD2 are poorly understood. Using transiently transfected human embryonic kidney (HEK)-293 cells as a model system, we report in the present study that PLD2 overexpressed in HEK-293 cells exhibits regulatory properties similar to PLD1 when stimulated in response to insulin and phorbol ester. Co-expression of PLD1 or PLD2 with PKC alpha results in constitutive activation of both PLD isoforms, which cannot be further stimulated by insulin. Co-expression of PLD1 with phospholipase C (PLC)gamma has the same effect, while co-expression of PLD2 with PLC gamma allows PLD2 activity to be stimulated in an insulin-dependent manner. The PKC-specific inhibitors bisindolylmaleimide and Go 6976 abolish insulin-induced PLD2 activation in HEK-293 cells co-expressing the insulin receptor, PLC gamma and PLD2, confirming that not only PLD1, but PLD2 as well, is regulated in a PKC-dependent manner. Finally, we provide evidence that PKC alpha is constitutively associated with PLD2. In summary, we demonstrate that insulin treatment results in activation of both PLD1 and PLD2 in appropriate cell types when the appropriate upstream intermediate signalling components, i.e. PKC alpha and PLC gamma, are expressed at sufficient levels. PMID- 11042116 TI - Activation of protein kinase B/cAkt in hepatocytes is sufficient for the induction of expression of the gene encoding glucokinase. AB - Inhibitors of signalling pathways were used to dissect the mechanism of insulin action on expression of the gene encoding glucokinase in cultured rat hepatocytes. Wortmannin and LY 294002 completely prevented the insulin-induced increase in glucokinase mRNA seen in unhibited cells, indicating that the phosphoinositide 3-kinase module has a key role. A ligand inducible protein kinase B (PKB, also termed cAkt) fusion protein was expressed by using adenoviral transduction of hepatocytes in primary culture. The PKB activity of this protein was shown to be activated in transduced hepatocytes within 30 min of the addition of 4-hydroxytamoxifen and to stay high for 8 h, as a result of serine phosphorylation at position 473 of PKB. The increase in PKB activity was reflected in the hyperphosphorylation of phosphorylated, heat and acid stable regulated by insulin protein (PHAS-I; also termed 4E-BP1, for eukaryotic initiation factor 4E-binding protein 1), a protein involved in the regulation of translation initiation. These effects were comparable to the insulin-induced activation of endogenous PKB and phosphorylation of PHAS-I in non-transduced hepatocytes. The addition of tamoxifen to transduced hepatocytes resulted in an induction of glucokinase mRNA with kinetics and magnitude similar to those of insulin-induced mRNA accumulation. The effect of tamoxifen depended on stimulated PKB activity because it did not occur in hepatocytes that were transduced with a mutant PKB fusion protein that was refractory to activation with tamoxifen. These results establish that acute activation of PKB is sufficient to produce an insulin-like induction of glucokinase in isolated hepatocytes. Together with the inhibition by phosphoinositide 3-kinase inhibitors, they suggest that the activation of PKB might be critical in mediating the induction of glucokinase by insulin. In addition, experiments showed that PD98059 decreased by half the increase in glucokinase mRNA brought about by insulin, suggesting a contributory role of the mitogen-activated protein kinase cascade. PMID- 11042117 TI - Specific interactions among transmembrane 4 superfamily (TM4SF) proteins and phosphoinositide 4-kinase. AB - In earlier work we established that phosphoinositide 4-kinase (PI 4-kinase) may associate with transmembrane 4 superfamily (TM4SF, tetraspanin) proteins, but critical specificity issues were not addressed. Here we demonstrate that at least five different TM4SF proteins (CD9, CD63, CD81, CD151 and A15/TALLA1) can associate with a similar or identical 55 kDa type II PI 4-kinase. These associations were specific, since we found no evidence for other phosphoinositide kinases (e.g. phosphoinositide 3-kinase and phosphoinositide-4-phosphate 5 kinase) associating with TM4SF proteins, and many other TM4SF proteins (including CD82 and CD53) did not associate with PI 4-kinase. CD63-PI 4-kinase complexes were almost entirely intracellular, and thus are distinct from other TM4SF-PI 4 kinase complexes (e.g. involving CD9), which are largely located in the plasma membrane. These results suggest that a specific subset of TM4SF proteins may recruit PI 4-kinase to specific membrane locations, and thereby influence phosphoinositide-dependent signalling. PMID- 11042118 TI - Sequencing, functional expression and characterization of rat NTPDase6, a nucleoside diphosphatase and novel member of the ecto-nucleoside triphosphate diphosphohydrolase family. AB - We have isolated and characterized the cDNA encoding nucleoside triphosphate diphosphohydrolase 6 (NTPDase6), a novel member of the ecto-nucleoside triphosphate diphosphohydrolase family. The rat-brain-derived cDNA has an open reading frame of 1365 bp encoding a protein of 455 amino acid residues, a calculated molecular mass of 49971 Da and a predicted N-terminal hydrophobic sequence. It shares 86% sequence identity with the human CD39L2 sequence and 48% and 51% identity respectively with sequences of the two related human and murine nucleoside diphosphatases (CD39L4, NTPDase5/ER-UDPase). The mRNA was expressed in all tissues investigated, revealing two major transcripts with differing abundances. PCR analysis suggests a single open reading frame. A Myc-His-tagged NTPDase6 was expressed in Chinese hamster ovary (CHO) and PC12 cells for immunological analysis and protein isolation. The protein was contained in membrane fractions of transfected CHO cells and occurred in a soluble form in the cell culture supernatants. NTPDase6 preferentially hydrolysed nucleoside 5' diphosphates. With different substrates the order of activity was GDP>IDP>>UDP,CDP>>ADP. Nucleoside 5'-triphosphates were hydrolysed only to a minor extent and no hydrolysis of nucleoside 5'-monophosphates was observed. The enzyme was strongly and equally activated by Ca(2+) and Mg(2+) and had a K(m) for GDP of 211 microM. The immunohistochemical analysis of transfected CHO and PC12 cells suggests that NTPDase6 is associated with the Golgi apparatus and to a small extent also with the plasma membrane. The enzyme might support glycosylation reactions in the Golgi apparatus and, when released from cells, might catalyse the hydrolysis of extracellular nucleotides. PMID- 11042119 TI - Role of non-covalent enzyme-substrate interactions in the reaction catalysed by cellobiose phosphorylase from Cellulomonas uda. AB - Steady-state kinetic studies of the enzymic glucosyl transfer to and from phosphate catalysed by cellobiose phosphorylase from Cellulomonas uda have shown that this enzyme operates by a ternary-complex kinetic mechanism in which beta cellobiose binds before phosphate, and beta-D-glucose and alpha-D-glucopyranosyl phosphate are released in that order. alpha-D-Glucopyranosyl fluoride (but not beta-D-glucopyranosyl fluoride) serves as alternative glucosyl donor for beta cellobiose synthesis with a specificity constant that is one-ninth that of the corresponding enzymic reaction with alpha-D-glucopyranosyl phosphate (approximately 20000 M(-1).s(-1) at 30 degrees C). The kinetic parameters for a complete series of deoxy and deoxyfluoro analogues of D-glucose have been determined and the data yield estimates of the net strengths of hydrogen-bonding interactions with the non-reacting hydroxy groups of D-glucose at the transition state (0.8-4.0 kcal/mol, where 1 cal identical with 4.184 J) and enable the prediction of the polarities of these hydrogen bonds. Each hydroxy group functions as donor of a hydrogen for bonding to probably a charged (at 3-OH) or neutral (at 2-OH and 6-OH) acceptor group on the enzyme. The equatorial 1-OH is essential for enzyme activity. Derivatives of D-glucose in which the 1-OH or the reacting 4-OH were replaced by hydrogen or fluorine have been tested as inhibitors to measure their affinities for the sugar-binding subsite +1 (numbered from the bond-cleaving/forming site). The data show that hydrogen-bonding interactions between the 1-OH and 4-OH and charged groups on the enzyme stabilize the ground-state ternary complex of the enzymic synthesis of beta-cellobiose by 2.3 and 0.4 kcal/mol, respectively, and assist the precise positioning of beta-D glucose for catalysis. PMID- 11042120 TI - Characterization of the mouse dynamin I gene promoter and identification of sequences that direct expression in neuronal cells. AB - Dynamin I is expressed at high levels in brain and its expression is regulated during the developmental stages of brain. To elucidate the molecular mechanism by which the expression is tissue-specifically regulated, we cloned the 5'-flanking region of the mouse dynamin I gene and determined the nucleotide sequence of 1036 bases upstream from the translation start site. Transient transfection studies with a chloramphenicol acetyltransferase reporter gene in neuroblastoma NS20Y and Lewis lung cells demonstrated that the 5'-flanking region has a cell-type specific promoter activity. Deletion analyses demonstrated that the minimal promoter activity was detected in the proximal region 195 bp upstream of the translation initiation codon (-90 to +105). The minimal promoter was embedded in a GC-rich region (75% GC content), in which an Sp1-binding motif and a nuclear factor (NF)-kappa B-like element (NE-1) were found, but it lacked TATA and CAAT boxes. Mutational analysis and electrophoretic mobility-shift assay analysis revealed that Sp1 binds to the Sp1 site and that this element is critical for the promoter activity of the dynamin I gene. We found that the NE-1 sequence is required for the expression of the dynamin I gene but NEBP (NE-1-binding protein), which binds to the NE-1 sequence, is not NF-kappa B. We also found that one base in the NE-1 sequence (the underlined G residue in GGGATTCGCGGA) is critical for binding specificity to discriminate between NEBP and NF-kappa B. By UV cross-linking analysis, we found that NEBP is an approx. 104 kDa nuclear protein. PMID- 11042122 TI - Cysteine residues in the C-terminus of the neutral- and basic-amino-acid transporter heavy-chain subunit contribute to functional properties of the system b(0,+)-type amino acid transporter. AB - The neutral- and basic-amino-acid-transport glycoprotein NBAT (rBAT, D2) expressed in renal and jejunal brush-border membranes interacts with the b(0,+)AT permease to produce a heteromeric transporter effecting amino acid and cystine absorption. NBAT mutations result in type I cystinuria. The b(0,+)AT permease is presumed to be the catalytic subunit, but we have been investigating the possibility that cysteine residues within the C-terminus of NBAT are also important for expression of transport function. NBAT mutants were produced with combinations of Cys(664/671/683)-->Ala substitutions. Mutants with Cys(664)-->Ala show decreased arginine and cystine transport and specifically lose sensitivity to inhibition of transport by the thiol-group reagent N-ethylmaleimide (NEM). We suggest that the C-terminus of NBAT may have a direct role in the mechanism of System b(0,+) transport (the major transport activity defective in type I cystinuria) and that Cys(664) of NBAT is the major target for NEM-induced inactivation of the transport mechanism. PMID- 11042121 TI - Platelet alpha IIb-beta 3 integrin engagement induces the tyrosine phosphorylation of Cbl and its association with phosphoinositide 3-kinase and Syk. AB - Agonist-induced platelet activation triggers 'inside-out' signalling which activates alpha IIb-beta 3, the most abundant integrin in platelet membranes. The engagement of activated alpha IIb-beta 3 integrin by linking fibrinogen is necessary for platelet aggregation, and this induces subsequent outside-in signalling, which enhances platelet activation. Here we studied the involvement of Cbl during alpha IIb-beta 3-integrin-mediated signal transduction. During thrombin-induced platelet activation, Cbl was tyrosine phosphorylated, and phosphoinositide 3-kinase (PI 3-kinase) activity measured in Cbl immunoprecipitates was increased. Both Cbl phosphorylation and its association with PI 3-kinase were dependent on alpha IIb-beta 3 engagement by linking fibrinogen. The P256 and anti-LIBS6 (ligand-induced binding site 6) antibodies, which activate platelets directly through alpha IIb-beta 3, induced Cbl phosphorylation and increased the PI 3-kinase activity associated with Cbl. Both thrombin and antibodies to alpha IIb-beta 3 induced association of Cbl with the tyrosine kinase, Syk. Experiments performed with inhibitors of tyrosine kinases indicated that both Src-family kinases and Syk contribute to phosphorylation of Cbl and its consequent association with PI 3-kinase. The results show that, following integrin alpha IIb-beta 3 engagement, Cbl is tyrosine phosphorylated, recruits PI 3-kinase to this integrin signalling pathway and possibly enhances PI 3-kinase activity, downstream of Src-family tyrosine kinases and Syk activation. PMID- 11042123 TI - Different receptors use inositol trisphosphate to mobilize Ca(2+) from different intracellular pools. AB - In cells expressing different receptors linked to Ins(1,4,5)P(3) formation, maximal stimulation of any one of them often releases all the Ins(1,4,5)P(3) sensitive Ca(2+) stores, suggesting that Ins(1,4, 5)P(3) is used similarly by many receptors. In single HEK-293 cells, ATP and carbamylcholine (CCh) stimulated Ca(2+) release from intracellular stores via a pathway that was entirely dependent on Ins(1,4,5)P(3). After stimulation with maximal concentrations of ATP or CCh in Ca(2+)-free medium, there was no response to a second stimulation with the same agonist, indicating that each agonist had emptied the Ins(1,4,5)P(3) sensitive stores to which it had access. However, the Ca(2+) release evoked by the second agonist was unaffected by prior stimulation with the first. We conclude that Ins(1,4,5)P(3) mediates the effects of both receptors, but Ins(1,4, 5)P(3) is more versatile than hitherto supposed, because the spatial organization of the signalling pathways apparently allows Ins(1,4, 5)P(3) made in response to each agonist to interact with different Ins(1,4,5)P(3) receptors. PMID- 11042124 TI - Regulation of Nramp1 mRNA stability by oxidants and protein kinase C in RAW264.7 macrophages expressing Nramp1(Gly169). AB - The murine Nramp1 (natural-resistance-associated macrophage protein) locus confers innate resistance against intracellular macrophage pathogens. The gene encodes a transporter molecule, which is rapidly recruited to the phagosome. Nramp1 functions as an iron transporter by transporting iron into the phagosome. Within the phagosome iron mediates anti-microbial killing by hydroxyl radical formation through the iron-catalysed Fenton/Haber-Weiss reaction. In addition to its effects on the growth of intracellular pathogens, Nramp1 exerts a wide range of pleiotropic effects in activated macrophages. One of these pleiotropic effects is the increased stability of a variety of mRNA species, including Nramp1 mRNA. In the present study, the stability of Nramp1 mRNA in Mycobacterium avium infected RAW264. 7 macrophages expressing either the Nramp1(Gly169) resistant allele or the Nramp1(Asp169) susceptible allele was examined. Nramp1 mRNA stability was greater in Nramp1(Gly169) macrophages than in Nramp1(Asp169) macrophages. The increase in Nramp1 mRNA stability in resistant macrophages was inhibited by antioxidants and protein kinase C (PKC) inhibitors, suggesting that Nramp1 mRNA stability is regulated by an oxidant-generated signalling pathway that requires PKC activity. This was corroborated by treating Nramp1(Asp169) macrophages with menadione, which generates reactive oxygen species within cells. Menadione increased Nramp1 mRNA stability to the level observed in resistant macrophages; this increase was also inhibited by a PKC inhibitor. Further, PKC activity was found to be greater in M. avium-infected Nramp1(Gly169) macrophages than in infected Nramp1(Asp169) macrophages and inhibited by treatment with an antioxidant. PMID- 11042125 TI - Analysis of a nucleotide-binding site of 5-lipoxygenase by affinity labelling: binding characteristics and amino acid sequences. AB - 5-Lipoxygenase (5LO) catalyses the first two steps in the biosynthesis of leukotrienes, which are inflammatory mediators derived from arachidonic acid. 5LO activity is stimulated by ATP; however, a consensus ATP-binding site or nucleotide-binding site has not been found in its protein sequence. In the present study, affinity and photoaffinity labelling of 5LO with 5'-p fluorosulphonylbenzoyladenosine (FSBA) and 2-azido-ATP showed that 5LO bound to the ATP analogues quantitatively and specifically and that the incorporation of either analogue inhibited ATP stimulation of 5LO activity. The stoichiometry of the labelling was 1.4 mol of FSBA/mol of 5LO (of which ATP competed with 1 mol/mol) or 0.94 mol of 2-azido-ATP/mol of 5LO (of which ATP competed with 0.77 mol/mol). Labelling with FSBA prevented further labelling with 2-azido-ATP, indicating that the same binding site was occupied by both analogues. Other nucleotides (ADP, AMP, GTP, CTP and UTP) also competed with 2-azido-ATP labelling, suggesting that the site was a general nucleotide-binding site rather than a strict ATP-binding site. Ca(2+), which also stimulates 5LO activity, had no effect on the labelling of the nucleotide-binding site. Digestion with trypsin and peptide sequencing showed that two fragments of 5LO were labelled by 2-azido ATP. These fragments correspond to residues 73-83 (KYWLNDDWYLK, in single-letter amino acid code) and 193-209 (FMHMFQSSWNDFADFEK) in the 5LO sequence. Trp-75 and Trp-201 in these peptides were modified by the labelling, suggesting that they were immediately adjacent to the C-2 position of the adenine ring of ATP. Given the stoichiometry of the labelling, the two peptide sequences of 5LO were probably near each other in the enzyme's tertiary structure, composing or surrounding the ATP-binding site of 5LO. PMID- 11042126 TI - Negative regulation of cytosolic phospholipase A(2) by melatonin in the rat pineal gland. AB - In this paper evidence that supports a new role for melatonin as a negative endogenous regulator of cytosolic phospholipase A(2) (cPLA(2)) is presented. When rat pineal glands were incubated in culture, time-dependent release of arachidonic acid (AA) was observed, which was significantly inhibited by a known 85-kDa cPLA(2) inhibitor, methyl arachidonyl fluorophosphonate. Co-incubation with melatonin inhibited the AA release in a concentration-dependent manner, and this decrease was accompanied by a reduction of cPLA(2) protein and mRNA expression. Melatonin-receptor agonists, 2-iodo-N-butanoyl-5-methoxytryptamine and 5-methoxycarbonylamino-N-acetyltryptamine, also decreased AA release and cPLA(2) protein and mRNA levels, while pre-incubation with the melatonin receptor antagonists luzindole and 2-phenylmelatonin abolished the melatonin effect. In vivo, as melatonin production reflected a typical diurnal variation, endogenous non-esterified AA and cPLA(2) mRNA levels in the rat pineal gland showed an off phase diurnal pattern in relation to melatonin levels. Intravenous administration of isoproterenol, which has been shown to elevate melatonin production, also decreased the levels of non-esterified AA and cPLA(2) mRNA significantly. Direct administration of melatonin to rats by intravenous injection decreased the levels of non-esterified AA, cPLA(2) protein and mRNA in rat pineal glands. In conclusion, melatonin endogenously down-regulates cPLA(2) expression, presumably through melatonin-receptor-mediated processes. PMID- 11042127 TI - Soluble GPI8 restores glycosylphosphatidylinositol anchoring in a trypanosome cell-free system depleted of lumenal endoplasmic reticulum proteins. AB - We previously established an in vitro assay for glycosylphosphatidylinositol (GPI) anchoring of proteins using trypanosome membranes. We now show that GPI anchoring is lost when the membranes are washed at high pH and restored to physiological pH prior to assay. We show that soluble component(s) of the endoplasmic reticulum that are lost in the high-pH wash are required for GPI anchoring. We reconstituted the high-pH extract with high-pH-treated membranes and demonstrated restoration of activity. Size fractionation of the high-pH extract indicated that the active component(s) was 30-50 kDa in size and was inactivated by iodoacetamide. Activity could also be restored by reconstituting the inactivated membranes with Escherichia coli-expressed, polyhistidine-tagged Leishmania mexicana GPI8 (GPI8-His; L. mexicana GPI8 is a soluble homologue of yeast and mammalian Gpi8p). No activity was seen when iodoacetamide-treated GPI8 His was used; however, GPI8-His could restore activity to iodoacetamide-treated membranes. Antibodies raised against L. mexicana GPI8 detected a protein of approx. 38 kDa in an immunoblot of the high-pH extract of trypanosome membranes. Our data indicate (1) that trypanosome GPI8 is a soluble lumenal protein, (2) that the interaction between GPI8 and other putative components of the transamidase may be dynamic, and (3) that GPI anchoring can be biochemically reconstituted using an isolated transamidase component. PMID- 11042128 TI - Ionization characteristics and chemical influences of aspartic acid residue 158 of papain and caricain determined by structure-related kinetic and computational techniques: multiple electrostatic modulators of active-centre chemistry. AB - The pK(a) of (Asp(158))-CO(2)H of papain (EC 3.4.22.2) was determined as 2.8 by using 4-chloro-7-nitrobenzofurazan (Nbf-Cl) as a reactivity probe targeted on the thiolate anion component of the Cys(25)/His(159) nucleophilic-acid/base motif of the catalytic site. The possibility of using Nbf-Cl for this purpose was established by modelling the papain-Nbf-Cl Meisenheimer intermediate by using QUANTA/CHARMM and performing molecular orbital calculations with MOPAC interfaced with Cerius 2. A pH-dependent stopped-flow kinetic study of the reaction of papain with Nbf-Cl established that the striking rate maximum at pH 3 results from reaction in a minor ionization state comprising (Cys(25))-S(-)/(His(159)) Im(+)H (in which Im represents imidazole) produced by protonic dissociation of (Cys(25))-SH/(His(159))-Im(+)H with pK(a) 3.3 and (Asp(158))-CO(2)H. Although the analogous intermediate in the reaction of caricain (EC 3.4.22.30) with Nbf-Cl has similar geometry, the pH-k profile (k being the second-order rate constant) lacks a rate maximum under acidic conditions. This precludes the experimental determination of the pK(a) value of (Asp(158))-CO(2)H of caricain, which was calculated to be 2.0 by solving the linearized Poisson-Boltzmann equation with the program UHBD ('University of Houston Brownian dynamics'). A value lower than 2.8 had been predicted by consideration of the hydrogen-bonded networks involving Asp(158) and its microenvironments in both enzymes. The difference between these pK(a) values (values not previously detected in reactions of either enzyme) accounts for the lack of the rate maximum in the caricain reaction and for the differences in the electronic absorption spectra of the two S-Nbf-enzymes under acidic conditions. The concept of control of cysteine proteinase activity by multiple electrostatic modulators, including (Asp(158))-CO(2)(-), which modifies traditional mechanistic views, is discussed. PMID- 11042129 TI - Cloning and expression of the human transient receptor potential 4 (TRP4) gene: localization and functional expression of human TRP4 and TRP3. AB - Mammalian homologues of the Drosophila transient receptor potential (TRP) protein have been proposed to function as ion channels, and in some cases as store operated or capacitative calcium entry channels. However, for each of the mammalian TRP proteins, different laboratories have reported distinct modes of cellular regulation. In the present study we describe the cloning and functional expression of the human form of TRP4 (hTRP4), and compare its activity with another well studied protein, hTRP3. When hTRP4 was transiently expressed in human embryonic kidney (HEK)-293 cells, basal bivalent cation permeability (barium) was increased. Whole-cell patch-clamp studies of hTRP4 expressed in Chinese hamster ovary cells revealed a constitutively active non-selective cation current which probably underlies the increased bivalent cation entry. Barium entry into hTRP4-transfected HEK-293 cells was not further increased by phospholipase C (PLC)-linked receptor activation, by intracellular calcium store depletion with thapsigargin, or by a synthetic diacylglycerol, 1-oleoyl-2-acetyl sn-glycerol (OAG). In contrast, transient expression of hTRP3 resulted in a bivalent cation influx that was markedly increased by PLC-linked receptor activation and by OAG, but not by thapsigargin. Despite the apparent differences in regulation of these two putative channel proteins, green fluorescent protein fusions of both molecules localized similarly to the plasma-membrane, notably in discrete punctate regions suggestive of specialized signalling complexes. Our findings indicate that while both hTRP4 and hTRP3 can apparently function as cation channels, their putative roles as components of capacitative calcium entry channels are not readily demonstrable by examining their behaviour when exogenously expressed in cells. PMID- 11042130 TI - The effect of peroxisome-proliferator-activated receptor-alpha on the activity of the cholesterol 7 alpha-hydroxylase gene. AB - Cholesterol 7 alpha-hydroxylase (Cyp7a1) plays a central role in the regulation of bile acid and cholesterol metabolism, and transcription of the gene is controlled by bile acids and hormones acting through a complex interaction with a number of potential steroid-hormone-binding sites. Transcriptional activity of the human CYP7A1 gene promoter transfected into HepG2 cells was decreased in a concentration-dependent manner by co-transfection with an expression vector for peroxisome-proliferator-activated receptor-alpha (PPAR alpha). This effect was augmented by 9-cis-retinoic acid receptor-alpha (RXR alpha) and activators of PPAR alpha to give a maximum inhibition of approx. 80%. The region responsible for this inhibition contained a site known to bind hepatocyte nuclear factor 4 (HNF4), and mutation of this site greatly decreased the effect. Co-expression of HNF4 increased promoter activity and decreased the effect of PPAR alpha. Gel mobility-shift assays failed to detect any binding of PPAR alpha/RXR alpha dimers to any regions of the promoter containing potential binding sites. Also the hepatic abundance of Cyp7a1 mRNA in mice in which the PPAR alpha gene was disrupted was the same as in normal mice, both during the dark phase, when the animals were feeding, and during the light phase, when mRNA abundance was greatly increased. Cholesterol feeding produced the same increase in hepatic Cyp7a1 mRNA abundance in PPAR alpha-null animals as in normals. It is concluded that, whereas PPAR alpha can affect CYP7A1 gene transcription in vitro through an indirect action, probably by competing for co-factors, this is unlikely to be a major influence on Cyp7a1 activity under normal physiological conditions. PMID- 11042131 TI - Gene expression of the dibasic-pair cleaving enzyme NRD convertase (N-arginine dibasic convertase) is differentially regulated in the GH3 pituitary and Mat-Lu prostate cell lines. AB - NRD convertase (N-arginine dibasic convertase, NRD-C) is a dibasic selective metalloprotease which cleaves on the N-terminal side of an arginine residue in a dibasic pair. Abundant in endocrine tissues, the highest levels are found in testis. The mechanism whereby NRD-C expression is regulated at the transcriptional level has been examined by reporter-gene assay and electrophoretic-mobility-shift assays. Analysis of the rat and human promoters show that they are highly conserved, containing a number of motifs which may correspond to transcription-factor binding sites. The rat promoter has been cloned into a luciferase reporter vector and analysed in a number of cell lines. Full functionality of the promoter is observed with 5' deletions to 411 bp upstream of the transcriptional start site in spermatid, prostate and pituitary cell lines. Further deletion to 101 bp causes a complete loss of activity in spermatid and prostate lines. By contrast, GH3 pituitary cells display no reduction in promoter activity with deletion to 101 bp of upstream sequence. A number of transcription-factor binding sites have been identified by electrophoretic-mobility-shift assays in the region 411-101; however, no differences in binding between the cell lines were observed. PMID- 11042132 TI - Novel consensus sequence for the Golgi apparatus casein kinase, revealed using proline-rich protein-1 (PRP1)-derived peptide substrates. AB - Previous studies have shown that the Golgi apparatus casein kinase (G-CK) recognizes phosphoacceptor sites specified by the triplet SXE/Sp, which is found in several phosphoproteins, besides casein itself. In the present study, we report that G-CK can phosphorylate, with comparable efficiency, sequences surrounding Ser-22 of salivary proline-rich protein-1 (PRP1), which do not conform to the SXE/Sp motif. By using a series of peptide substrates derived from the PRP1 Ser-22 site, we also have shown that the optimal consensus sequence recognized by G-CK in this case was SXQXX(D/E)3, where the acidic residues at positions n+5 to n+7 and, to a lesser extent, the glutamine residue at position n+2 are the critical determinants. PMID- 11042133 TI - Histone carbonylation in vivo and in vitro. AB - Non-enzymic damage to nuclear proteins has potentially severe consequences for the maintenance of genomic integrity. Introduction of carbonyl groups into histones in vivo and in vitro was assessed by Western blot immunoassay and reductive incorporation of tritium from radiolabelled NaBH(4) (sodium borohydride). Histone H1 extracted from bovine thymus, liver and spleen was found to contain significantly elevated amounts of protein-bound carbonyl groups as compared with core histones. The carbonyl content of nuclear proteins of rat pheochromocytoma cells (PC12 cells) was not greatly increased following oxidative stress induced by H(2)O(2), but was significantly increased following alkylating stress induced by N-methyl-N'-nitro-N-nitrosoguanidine or by combined oxidative and alkylating stress. Free ADP-ribose, a reducing sugar generated in the nucleus in proportion to DNA strand breaks, was shown to be a potent histone H1 carbonylating agent in isolated PC12 cell nuclei. Studies of the mechanism of histone H1 modification by ADP-ribose indicate that carbonylation involves formation of a stable acyclic ketoamine. Our results demonstrate preferential histone H1 carbonylation in vivo, with potentially important consequences for chromatin structure and function. PMID- 11042134 TI - Very low surfactant protein C contents in newborn Belgian White and Blue calves with respiratory distress syndrome. AB - We have studied a respiratory distress syndrome (RDS) occurring in newborn calves of the Belgian White and Blue (BWB) breed that represents the large majority of beef cattle in Belgium. Pulmonary surfactant isolated from 14 BWB newborn calves that died from RDS and from 7 healthy controls was analysed for composition and surface activity. An extremely low content or, in some instances, an absence of surfactant protein C (SP-C) was detected in the RDS samples by Western blotting and differential amino acid analysis [0.03+/-0.01% (w/w) relative to total phospholipids, compared with 0.39+/-0.06% for healthy controls (means+/-S.E.M., P < 0.001)]. The contents of surfactant protein B (SP-B) were similar in RDS and control samples. The crude surfactant samples isolated from RDS calves had higher ratios of total protein to total phospholipid, altered phospholipid profiles and lower SP-A contents. Both crude and organic extracts of RDS surfactant samples showed increased dynamic surface tension compared with healthy controls when evaluated with a pulsating-bubble surfactometer. The addition of purified SP-C to organic extracts of RDS surfactant samples lowered surface tension. Strongly decreased levels of mature SP-C associated with fatal RDS and altered surface activity in vitro have, to the best of our knowledge, not been previously reported. The mechanisms underlying RDS and the decrease in SP-C in BWB calves remain to be established. PMID- 11042135 TI - Time-dependent activation of the semicarbazide-sensitive amine oxidase (SSAO) from ox lung microsomes. AB - The activity of ox lung microsomal semicarbazide-sensitive amine oxidase (EC 1.4.3.6; SSAO) towards benzylamine increased 20-fold during incubation at 37 degrees C. After an initial lag-period, activation was first-order with time and complete after approx. 20 h. No significant changes in activity towards methylamine, histamine or 2-phenylethylamine were observed, although mixed substrate experiments were consistent with the same enzyme being involved in the oxidation of all these substrates, both before and after time-dependent activation. The enzyme-tryptophan fluorescence increased on incubation at 37 degrees C in parallel with the increase in activity towards benzylamine. Treatment of the activated-enzyme preparation with 6 M guanidinium chloride followed by dialysis, caused both the activity towards benzylamine and the fluorescence to fall to that occurring before activation. However, incubation of this preparation at 37 degrees C resulted in increases in fluorescence and activity similar to those seen with the unactivated enzyme. Benzylamine oxidation was inhibited, uncompetitively with respect to oxygen, by high substrate concentrations but no such inhibition was observed with the other amines. Activation resulted in an increase in V(max) for benzylamine oxidation, with no significant alterations in the K(m) or the K(si) for high-substrate inhibition. Kinetic studies were consistent with sequential mechanisms being followed for the oxidation of both benzylamine and methylamine but the dependence on oxygen concentration was complex. These results might indicate that benzylamine follows a different reaction pathway from the other substrates, with substrate-specific activation involving a reaction step that is rate-limiting for benzylamine oxidation but not for the others. PMID- 11042136 TI - Bis(monoacylglycerol) phosphate in rat uterine stromal cells: structural characterization and specific esterification of docosahexaenoic acid. AB - In rat uterine stromal cells (U(III) cells), docosahexaenoic acid (DHA) was esterified extensively in alkenylacyl-glycerophosphoethanolamine and in an unknown phospholipid accounting for only 0.7% of the total phospholipid. The latter was identified as a bis(monoacylglycerol) phosphate (BMP) using MS. Incorporation studies using C(18:3)n-3 and C(20:5)n-3 demonstrated that BMP had a high specificity to incorporate DHA and C(22) polyunsaturated fatty acids of the (n-3) series. By contrast, polyunsaturated fatty acids of the (n-6) series were never incorporated into BMP. Incubation of U(III) cells with 5 microM DHA for 24 h increased the DHA content of BMP from 36 to 71% of the total acyl chains. [(3)H]DHA-labelled BMP purified as a single TLC spot was resolved into three peaks using HPLC. These peaks were also observed when cells were labelled with [(3)H]phosphatidylglycerol, an exogenous BMP precursor, and with [(33)P]P(i). Electrospray MS of BMP from control cells showed that the first two peaks contained the same molecular species (mainly C(22:6)n-3/C(22:6)n-3 and C(18:1)n 9/C(22:6)n-3) while the third peak mainly contained the C(18:1)n-9/C(18:1)n-9 species. The stereoconfiguration analysis of the compounds revealed an sn-glycero 3-phospho-1'-sn-glycerol configuration for the first peak and sn-glycero-1 phospho-1'-sn-glycerol configurations for the other two. BMP from rat testis was used to establish the positions of the acyl groups. More than 70% of its acyl chains were C(22:5) n-6. It was separated on HPLC into three peaks that co migrated with the three peaks of BMP from U(III) cells. Lipase activity and NMR analysis of the second peak showed that fatty acids esterified the primary alcohol group on each glycerol moiety. We conclude that the three peaks are stereoisomeric compounds with different acyl-chain locations and may be the result of different metabolic fates depending on subcellular localization. PMID- 11042137 TI - Endurance training increases stimulation of uncoupling of skeletal muscle mitochondria in humans by non-esterified fatty acids: an uncoupling-protein mediated effect? AB - Uncoupled respiration (UCR) is an essential property of muscle mitochondria and has several functions in the cell. We hypothesized that endurance training may alter the magnitude and properties of UCR in human muscle. Isolated mitochondria from muscle biopsies taken before and after 6 weeks of endurance exercise training (n=8) were analysed for UCR. To investigate the role of uncoupling protein 2 (UCP2) and UCP3 in UCR, the sensitivity of UCR to UCP-regulating ligands (non-esterified fatty acids and purine nucleotides) and UCP2 and UCP3 mRNA expression in muscle were examined. Oleate increased the mitochondrial oxygen consumption rate, an effect that was not attenuated by GDP and/or cyclosporin A. The effect of oleate was significantly greater after compared with before training. Training had no effect on UCP2 or UCP3 mRNA levels, but after training the relative increase in respiration rate induced by oleate was positively correlated with the UCP2 mRNA level. In conclusion, we show that the sensitivity of UCR to non-esterified fatty acids is up-regulated by endurance training. This suggests that endurance training causes intrinsic changes in mitochondrial function, which may enhance the potential for regulation of aerobic energy production, prevent excess free radical generation and contribute to a higher basal metabolic rate. PMID- 11042138 TI - Shared control of hepatic glycogen synthesis by glycogen synthase and glucokinase. AB - We have used recombinant adenoviruses (AdCMV-RLGS and AdCMV-GK) to overexpress the liver isoforms of glycogen synthase (GS) and glucokinase (GK) in primary cultured rat hepatocytes. Glucose activated overexpressed GS in a dose-dependent manner and caused the accumulation of larger amounts of glycogen in the AdCMV RLGS-treated hepatocytes. The concentration of intermediate metabolites of the glycogenic pathway, such as glucose 6-phosphate (Glc-6-P) and UDP-glucose, were not significantly altered. GK overexpression also conferred on the hepatocyte an enhanced capacity to synthesize glycogen in response to glucose, as described previously [Seoane, Gomez-Foix, O'Doherty, Gomez-Ara, Newgard and Guinovart (1996) J. Biol. Chem. 271, 23756-23760], although, in this case, they accumulated Glc-6-P. When GS and GK were simultaneously overexpressed, the accumulation of glycogen was enhanced in comparison with cells overexpressing either GS or GK. Our results are consistent with the hypothesis that liver GS catalyses the rate limiting step of hepatic glycogen synthesis. However, hepatic glycogen deposition from glucose is submitted to a system of shared control in which the 'controller', GS, is, in turn, controlled by GK. This control is indirectly exerted through Glc-6-P, which 'switches on' GS dephosphorylation and activation. PMID- 11042139 TI - N-acyl-dopamines: novel synthetic CB(1) cannabinoid-receptor ligands and inhibitors of anandamide inactivation with cannabimimetic activity in vitro and in vivo. AB - We reported previously that synthetic amides of polyunsaturated fatty acids with bioactive amines can result in substances that interact with proteins of the endogenous cannabinoid system (ECS). Here we synthesized a series of N-acyl dopamines (NADAs) and studied their effects on the anandamide membrane transporter, the anandamide amidohydrolase (fatty acid amide hydrolase, FAAH) and the two cannabinoid receptor subtypes, CB(1) and CB(2). NADAs competitively inhibited FAAH from N18TG2 cells (IC(50)=19-100 microM), as well as the binding of the selective CB(1) receptor ligand, [(3)H]SR141716A, to rat brain membranes (K(i)=250-3900 nM). The arachidonoyl (20:4 omega 6), eicosapentaenoyl (20:5 omega 3), docosapentaenoyl (22:5 omega 3), alpha-linolenoyl (18:3 omega 3) and pinolenoyl (5c,9c,12c 18:3 omega 6) homologues were also found to inhibit the anandamide membrane transporter in RBL-2H3 basophilic leukaemia and C6 glioma cells (IC(50)=17.5-33 microM). NADAs did not inhibit the binding of the CB(1)/CB(2) receptor ligand, [(3)H]WIN55,212-2, to rat spleen membranes (K(i)>10 microM). N-arachidonyl-dopamine (AA-DA) exhibited 40-fold selectivity for CB(1) (K(i)=250 nM) over CB(2) receptors, and N-docosapentaenoyl-dopamine exhibited 4 fold selectivity for the anandamide transporter over FAAH. AA-DA (0.1-10 microM) did not displace D1 and D2 dopamine-receptor high-affinity ligands from rat brain membranes, thus suggesting that this compound has little affinity for these receptors. AA-DA was more potent and efficacious than anandamide as a CB(1) agonist, as assessed by measuring the stimulatory effect on intracellular Ca(2+) mobilization in undifferentiated N18TG2 neuroblastoma cells. This effect of AA-DA was counteracted by the CB(1) antagonist SR141716A. AA-DA behaved as a CB(1) agonist in vivo by inducing hypothermia, hypo-locomotion, catalepsy and analgesia in mice (1-10 mg/kg). Finally, AA-DA potently inhibited (IC(50)=0.25 microM) the proliferation of human breast MCF-7 cancer cells, thus behaving like other CB(1) agonists. Also this effect was counteracted by SR141716A but not by the D2 antagonist haloperidol. We conclude that NADAs, and AA-DA in particular, may be novel and useful probes for the study of the ECS. PMID- 11042140 TI - Expression of maize eukaryotic initiation factor (eIF) iso4E is regulated at the translational level. AB - Mechanisms for regulation of gene expression at the translational level have been reported at specific developmental stages in eukaryotes. Control of eukaryotic initiation factor (eIF) 4E availability by insulin/growth factors constitutes a main point of translational regulation. The aim of the present research was to understand the regulatory mechanism(s) behind the differential expression of two main 4E factors present in maize embryonic axes during germination. De novo synthesis of eIFiso4E initiates earlier and is faster than that of eIF4E in maize axes. Insulin addition to maize axes stimulated de novo synthesis of the eIFiso4E protein, but not that of eIF4E. Specific recruitment of the eIFiso4E transcript into polysomes was observed in these axes after insulin stimulation. Inhibitors of the insulin signal-transduction pathway, wortmannin and rapamycin, reversed the insulin effect. In vitro translation of maize poly(A)(+) RNAs by S6 ribosomal protein (rp)-phosphorylated ribosomes demonstrated a strong increase in eIFiso4E synthesis, as compared with its translation by S6 rp-non-phosphorylated ribosomes. Other mRNAs from the poly(A)(+) RNA set, including the eIF4E mRNA, did not show differential translation with regard to the S6-phosphorylated status of the ribosomes. The overall results indicate that eIFiso4E, but not eIF4E, cell content is regulated by de novo synthesis in maize axes during germination, most probably by specific mRNA recruitment into polysomes via a signal-transduction pathway involving S6 rp phosphorylation. PMID- 11042141 TI - Identification of Glu-519 as the catalytic nucleophile in beta-mannosidase 2A from Cellulomonas fimi. AB - Incubation of the beta-mannosidase Man2A from Cellulomonas fimi with 2-deoxy-2 fluoro-beta-D-mannosyl fluoride (2FMan beta F) resulted in time-dependent inactivation of the enzyme (inactivation rate constant k(i)=0.57 min(-1), dissociation constant for the inactivator K(i)=0.41 mM) through the accumulation of a covalent 2-deoxy-2-fluoro-alpha-D-mannosyl-beta-mannosidase 2A (2FMan-Man2A) enzyme intermediate, as observed by electrospray ionization mass spectrometry. The stoichiometry of inactivation was 1:1. Removal of excess inactivator and regeneration of active enzyme by transglycosylation of the covalently attached inhibitor to gentiobiose [Glc beta(1-6)Glc] demonstrated that the covalent intermediate was catalytically competent. Comparison by MS of the peptic digests of 2FMan-Man2A with peptic digests of native Man2A revealed a peptide of m/z 1520 that was unique to 2FMan-Man2A, and one of m/z 1036.5 that was unique to a Man2A peptide. Their sequences, determined by collision-induced fragmentation, were CSEFGFQGPPTW and FGFQGPPTW, corresponding to residues 517-528 and 520-528 of Man2A respectively. The difference in mass of 483.5 between the two peptides equals the sum of the masses of the tripeptide CSE plus that of 2-fluoromannose. It was concluded that in 2FMan-Man2A, the 2-fluoromannose esterified to Glu-519 blocks hydrolysis of the Glu-519-Phe-520 peptide bond, and that Glu-519 is the catalytic nucleophile in this enzyme. This residue is conserved in all members of family 2 of the glycosyl hydrolases. This represents the first ever labelling and identification of an active-site nucleophile in a beta-mannosidase. PMID- 11042142 TI - Of mice, men, and the genome. PMID- 11042143 TI - Linkage disequilibrium and the search for complex disease genes. PMID- 11042144 TI - HEAT repeats associated with condensins, cohesins, and other complexes involved in chromosome-related functions. AB - HEAT repeats correspond to tandemly arranged curlicue-like structures that appear to serve as flexible scaffolding on which other components can assemble. Using sensitive sequence analysis techniques we detected HEAT repeats in various chromosome-associated proteins, including four families of proteins associated with condensins and cohesins, which are nuclear complexes that contain structural maintenance of chromosome (SMC) proteins. Among the proteins detected were the XCAP-D2 and XCAP-G subunits of the Xenopus laevis 13S condensin complex, the Aspergillus BimD and Sordaria macrospora Spo76p proteins, the budding yeast Scc2p protein, and the related Drosophila transcriptional activator Nipped-B. Clathrin adaptor and COP-I coatomer subunits, which function in vesicle coat assembly and were previously noted to share weak sequence similarity to condensin subunits, also contain HEAT repeats. HEAT repeats were also found in the TBP-associated TIP120 protein, a global enhancer of transcription, and in the budding yeast Mot1p protein, which is a member of the SWI2/SNF2 family. SWI2/SNF2 proteins, some of which are helicases, perform diverse roles in transcription control, DNA repair, and chromosome segregation and form chromatin-remodeling complexes. HEAT repeats also were found in dis1-TOG family and cofactor D family microtubule associated proteins, which, owing to their roles in microtubule dynamics, perform functions related to mitotic progression and chromosome segregation. Hence, our analysis predicts structural features of these proteins and suggests that HEAT repeats may play important roles in chromosome dynamics. PMID- 11042145 TI - Organization of mouse Iroquois homeobox genes in two clusters suggests a conserved regulation and function in vertebrate development. AB - Iroquois proteins comprise a conserved family of homeodomain-containing transcription factors involved in patterning and regionalization of embryonic tissues in both vertebrates and invertebrates. Earlier studies identified four murine Iroquois (Irx) genes. Here we report the isolation of two additional members of the murine gene family, Irx5 and Irx6. Phylogenetic analysis of the Irx gene family revealed distinct clades for fly and vertebrate genes, and vertebrate members themselves were classified into three pairs of cognate genes. Mapping of the murine Irx genes identified two gene clusters located on mouse chromosomes 8 and 13, respectively. Each gene cluster is represented by three Irx genes whose relative positions within both clusters are strictly conserved. Combined results from phylogenetic, linkage, and physical mapping studies provide evidence for the evolution of two Irx gene clusters by duplication of a larger chromosomal region and dispersion to two chromosomal locations. The maintenance of two cognate Irx gene clusters during vertebrate evolution suggests that their genomic organization is important for the regulation, expression, and function of Irx genes during embryonic development. PMID- 11042146 TI - Chromosome evolution: the junction of mammalian chromosomes in the formation of mouse chromosome 10. AB - During evolution, chromosomes are rearranged and become fixed into new patterns in new species. The relatively conservative nature of this process supports predictions of the arrangement of ancestral mammalian chromosomes, but the basis for these rearrangements is unknown. Physical mapping of mouse chromosome 10 (MMU 10) previously identified a 380-kb region containing the junction of material represented in human on chromosomes 21 (HSA 21) and 22 (HSA 22) that occurred in the evolutionary lineage of the mouse. Here, acquisition of 275 kb of mouse genomic sequence from this region and comparative sequence analysis with HSA 21 and HSA 22 narrowed the junction from 380 kb to 18 kb. The minimal junction region on MMU 10 contains a variety of repeats, including an L32-like ribosomal element and low-copy sequences found on several mouse chromosomes and represented in the mouse EST database. Sequence level analysis of an interchromosomal rearrangement during evolution has not been reported previously. PMID- 11042147 TI - Biosynthesis of isoprenoids via mevalonate in Archaea: the lost pathway. AB - Isoprenoid compounds are ubiquitous in living species and diverse in biological function. Isoprenoid side chains of the membrane lipids are biochemical markers distinguishing archaea from the rest of living forms. The mevalonate pathway of isoprenoid biosynthesis has been defined completely in yeast, while the alternative, deoxy-D-xylulose phosphate synthase pathway is found in many bacteria. In archaea, some enzymes of the mevalonate pathway are found, but the orthologs of three yeast proteins, accounting for the route from phosphomevalonate to geranyl pyrophosphate, are missing, as are the enzymes from the alternative pathway. To understand the evolution of isoprenoid biosynthesis, as well as the mechanism of lipid biosynthesis in archaea, sequence motifs in the known enzymes of the two pathways of isoprenoid biosynthesis were analyzed. New sequence relationships were detected, including similarities between diphosphomevalonate decarboxylase and kinases of the galactokinase superfamily, between the metazoan phosphomevalonate kinase and the nucleoside monophosphate kinase superfamily, and between isopentenyl pyrophosphate isomerases and MutT pyrophosphohydrolases. Based on these findings, orphan members of the galactokinase, nucleoside monophosphate kinase, and pyrophosphohydrolase families in archaeal genomes were evaluated as candidate enzymes for the three missing steps. Alternative methods of finding these missing links were explored, including physical linkage of open reading frames and patterns of ortholog distribution in different species. Combining these approaches resulted in the generation of a short list of 13 candidate genes for the three missing functions in archaea, whose participation in isoprenoid biosynthesis is amenable to biochemical and genetic investigation. PMID- 11042148 TI - Potential gene conversion and source genes for recently integrated Alu elements. AB - Alu elements comprise >10% of the human genome. We have used a computational biology approach to analyze the human genomic DNA sequence databases to determine the impact of gene conversion on the sequence diversity of recently integrated Alu elements and to identify Alu elements that were potentially retroposition competent. We analyzed 269 Alu Ya5 elements and identified 23 members of a new Alu subfamily termed Ya5a2 with an estimated copy number of 35 members, including the de novo Alu insertion in the NF1 gene. Our analysis of Alu elements containing one to four (Ya1-Ya4) of the Ya5 subfamily-specific mutations suggests that gene conversion contributed as much as 10%-20% of the variation between recently integrated Alu elements. In addition, analysis of the middle A-rich region of the different Alu Ya5 members indicates a tendency toward expansion of this region and subsequent generation of simple sequence repeats. Mining the databases for putative retroposition-competent elements that share 100% nucleotide identity to the previously reported de novo Alu insertions linked to human diseases resulted in the retrieval of 13 exact matches to the NF1 Alu repeat, three to the Alu element in BRCA2, and one to the Alu element in FGFR2 (Apert syndrome). Transient transfections of the potential source gene for the Apert's Alu with its endogenous flanking genomic sequences demonstrated the transcriptional and presumptive transpositional competency of the element. PMID- 11042149 TI - Reading between the LINEs: human genomic variation induced by LINE-1 retrotransposition. AB - The insertion of mobile elements into the genome represents a new class of genetic markers for the study of human evolution. Long interspersed elements (LINEs) have amplified to a copy number of about 100,000 over the last 100 million years of mammalian evolution and comprise approximately 15% of the human genome. The majority of LINE-1 (L1) elements within the human genome are 5' truncated copies of a few active L1 elements that are capable of retrotransposition. Some of the young L1 elements have inserted into the human genome so recently that populations are polymorphic for the presence of an L1 element at a particular chromosomal location. L1 insertion polymorphisms offer several advantages over other types of polymorphisms for human evolution studies. First, they are typed by rapid, simple, polymerase chain reaction (PCR)-based assays. Second, they are stable polymorphisms that rarely undergo deletion. Third, the presence of an L1 element represents identity by descent, because the probability is negligible that two different young L1 repeats would integrate independently between the exact same two nucleotides. Fourth, the ancestral state of L1 insertion polymorphisms is known to be the absence of the L1 element, which can be used to root plots/trees of population relationships. Here we report the development of a PCR-based display for the direct identification of dimorphic L1 elements from the human genome. We have also developed PCR-based assays for the characterization of six polymorphic L1 elements within the human genome. PCR analysis of human/rodent hybrid cell line DNA samples showed that the polymorphic L1 elements were located on several different chromosomes. Phylogenetic analysis of nonhuman primate DNA samples showed that all of the recently integrated "young" L1 elements were restricted to the human genome and absent from the genomes of nonhuman primates. Analysis of a diverse array of human populations showed that the allele frequencies and level of heterozygosity for each of the L1 elements was variable. Polymorphic L1 elements represent a new source of identical-by-descent variation for the study of human evolution. [The sequence data described in this paper have been submitted to the GenBank data library under accession nos. AF242435-AF242451.] PMID- 11042150 TI - Molecular genetic maps in wild emmer wheat, Triticum dicoccoides: genome-wide coverage, massive negative interference, and putative quasi-linkage. AB - The main objectives of the study reported here were to construct a molecular map of wild emmer wheat, Triticum dicoccoides, to characterize the marker-related anatomy of the genome, and to evaluate segregation and recombination patterns upon crossing T. dicoccoides with its domesticated descendant Triticum durum (cultivar Langdon). The total map length exceeded 3000 cM and possibly covered the entire tetraploid genome (AABB). Clusters of molecular markers were observed on most of the 14 chromosomes. AFLP (amplified fragment length polymorphism) markers manifested a random distribution among homologous groups, but not among genomes and chromosomes. Genetic differentiation between T. dicoccoides and T. durum was attributed mainly to the B genome as revealed by AFLP markers. The segregation-distorted markers were mainly clustered on 4A, 5A, and 5B chromosomes. Homeoalleles, differentially conferring the vigor of gametes, might be responsible for the distortion on 5A and 5B chromosomes. Quasilinkage, deviation from free recombination between markers of nonhomologous chromosomes, was discovered. Massive negative interference was observed in most of the chromosomes (an excess of double crossovers in adjacent intervals relative to the expected rates on the assumption of no interference). The general pattern of distribution of islands of negative interference included near-centromeric location, spanning the centromere, and median/subterminal location. [An appendix describing the molecular marker loci is available as an online supplement at http://www.genome.org.] PMID- 11042151 TI - Sequence diversity and large-scale typing of SNPs in the human apolipoprotein E gene. AB - A common strategy for genotyping large samples begins with the characterization of human single nucleotide polymorphisms (SNPs) by sequencing candidate regions in a small sample for SNP discovery. This is usually followed by typing in a large sample those sites observed to vary in a smaller sample. We present results from a systematic investigation of variation at the human apolipoprotein E locus (APOE), as well as the evaluation of the two-tiered sampling strategy based on these data. We sequenced 5.5 kb spanning the entire APOE genomic region in a core sample of 72 individuals, including 24 each of African-Americans from Jackson, Mississippi; European-Americans from Rochester, Minnesota; and Europeans from North Karelia, Finland. This sequence survey detected 21 SNPs and 1 multiallelic indel, 14 of which had not been previously reported. Alleles varied in relative frequency among the populations, and 10 sites were polymorphic in only a single population sample. Oligonucleotide ligation assays (OLA) were developed for 20 of these sites (omitting the indel and a closely-linked SNP). These were then scored in 2179 individuals sampled from the same three populations (n = 843, 884, and 452, respectively). Relative allele frequencies were generally consistent with estimates from the core sample, although variation was found in some populations in the larger sample at SNPs that were monomorphic in the corresponding smaller core sample. Site variation in the larger samples showed no systematic deviation from Hardy-Weinberg expectation. The large OLA sample clearly showed that variation in many, but not all, of OLA-typed SNPs is significantly correlated with the classical protein-coding variants, implying that there may be important substructure within the classical epsilon 2, epsilon 3, and epsilon 4 alleles. Comparison of the levels and patterns of polymorphism in the core samples with those estimated for the OLA-typed samples shows how nucleotide diversity is underestimated when only a subset of sites are typed and underscores the importance of adequate population sampling at the polymorphism discovery stage. [The sequence data described in this paper have been submitted to the GenBank data library under accession no. AF261279.] PMID- 11042152 TI - Cloning and functional analysis of cDNAs with open reading frames for 300 previously undefined genes expressed in CD34+ hematopoietic stem/progenitor cells. AB - Three hundred cDNAs containing putatively entire open reading frames (ORFs) for previously undefined genes were obtained from CD34+ hematopoietic stem/progenitor cells (HSPCs), based on EST cataloging, clone sequencing, in silico cloning, and rapid amplification of cDNA ends (RACE). The cDNA sizes ranged from 360 to 3496 bp and their ORFs coded for peptides of 58-752 amino acids. Public database search indicated that 225 cDNAs exhibited sequence similarities to genes identified across a variety of species. Homology analysis led to the recognition of 50 basic structural motifs/domains among these cDNAs. Genomic exon-intron organization could be established in 243 genes by integration of cDNA data with genome sequence information. Interestingly, a new gene named as HSPC070 on 3p was found to share a sequence of 105bp in 3' UTR with RAF gene in reversed transcription orientation. Chromosomal localizations were obtained using electronic mapping for 192 genes and with radiation hybrid (RH) for 38 genes. Macroarray technique was applied to screen the gene expression patterns in five hematopoietic cell lines (NB4, HL60, U937, K562, and Jurkat) and a number of genes with differential expression were found. The resource work has provided a wide range of information useful not only for expression genomics and annotation of genomic DNA sequence, but also for further research on the function of genes involved in hematopoietic development and differentiation. PMID- 11042153 TI - High-resolution transcript map of the region spanning D12S1629 and D12S312 at chromosome 12q13: triple A syndrome-linked region. AB - For those searching for human disease-causing genes, information on the position of genes with respect to genetic markers is essential. The physical map composed of ESTs and genetic markers provides the positional information of these markers as well as the starting point of gene identification in the form of genomic clones containing exons. To facilitate the effort of identification of genes in the region spanning D12S1629 and D12S312, we constructed a high-resolution transcript map with PAC/BAC/cosmid clones. The strategy for the construction of such a map involved utilization of STSs for the screening of the large insert bacterial chromosome libraries and a chromosome 12-specific cosmid library by hybridization. The contig was constructed based on the STS contents of the clones. The resulting high-resolution transcript map of the region between P273P14/SP6 and D12S312 spans 4.4 cM from 66.8 to 71.2 cM of the Genethon genetic map and represents approximately 2.4 Mb. It was composed of 81 BAC, 45 PAC, and 91 cosmid clones with a minimal tiling path consisting of 16 BAC and 4 PAC clones. These clones are being used to sequence this part of chromosome 12. We determined the order of 135 STSs including 74 genes and ESTs in the map. Among these, 115 STSs were unambiguously ordered, resulting in one ordered marker per 21 kb. The order of keratin type II locus genes was determined. This map would greatly enhance the positional cloning effort of the responsible genes for those diseases that are linked to this region, including male germ cell tumor as well as palmoplantar keratoderma, Bothnian-type, and triple A syndrome. This transcript map was localized at human chromosome 12q13. PMID- 11042154 TI - Maternal environment and genotype interact to establish diabesity in mice. AB - Obesity, a major risk factor for type II diabetes, is becoming more prevalent in Western populations consuming high calorie diets while expending less energy both at the workplace and at home. Most human obesity, and probably most type II diabetes as well, reflects polygenic rather than monogenic inheritance. We have genetically dissected a polygenic mouse model of obesity-driven type II diabetes by outcrossing the obese, diabetes-prone, NZO (New Zealand Obese)/HlLt strain to the relatively lean NON (Nonobese Nondiabetic)/Lt strain, and then reciprocally backcrossing obese F1 mice to the lean NON/Lt parental strain. A continuous distribution of body weights was observed in a population of 203 first backcross males. The 22% of first backcross males developing overt diabetes showed highest peripubertal weight gains and earliest development of hyperinsulinemia. We report a complex diabetes-predisposing ("diabesity") QTL (Quantitative Trait Loci) on chromosome 1 contributing significant main effects to increases in body weight, plasma insulin, and plasma glucose. NZO contributed QTL with significant main effects on adiposity parameters on chromosomes 12 and 5. A NON QTL on chromosome 14 interacted epistatically with the NZO obesity QTL on chromosome 12 to increase adiposity. Although the main effect of the diabetogenic QTL on chromosome 1 was on rapid growth rather than adiposity, it interacted epistatically with the obesity QTL on chromosome 12 to increase plasma glucose levels. Additional complex epistatic interactions eliciting significant increases in body weight and/or plasma glucose were found between the NZO-contributed QTL on chromosome 1 and other NZO-contributed QTL on chromosomes 15 and 17, as well as with an NON contributed QTL on chromosome 2. We further show that certain of these intergenic interactions are predicated on, or enhanced by, the maternal postparturitional environment. We show by cross-fostering experiments that the maternal environmental influence in part is because of the presence of early obesity inducing factors in the milk of obese F1 dams. We also discuss a strategy for using recombinant congenic strains to separate and reassemble interacting QTL for future study. PMID- 11042155 TI - SNP profile within the human major histocompatibility complex reveals an extreme and interrupted level of nucleotide diversity. AB - The human major histocompatibility complex (MHC) is characterized by polymorphic multicopy gene families, such as HLA and MIC (PERB11); duplications; insertions and deletions (indels); and uneven rates of recombination. Polymorphisms at the antigen recognition sites of the HLA class I and II genes and at associated neutral sites have been attributed to balancing selection and a hitchhiking effect, respectively. We, and others, have previously shown that nucleotide diversity between MHC haplotypes at non-HLA sites is unusually high (>10%) and up to several times greater than elsewhere in the genome (0.08%-0.2%). We report here the most extensive analysis of nucleotide diversity within a continuous sequence in the genome. We constructed a single nucleotide polymorphism (SNP) profile that reveals a pattern of extreme but interrupted levels of nucleotide diversity by comparing a continuous sequence within haplotypes in three genomic subregions of the MHC. A comparison of several haplotypes within one of the genomic subregions containing the HLA-B and -C loci suggests that positive selection is operating over the whole subgenomic region, including HLA and non HLA genes. [The sequence data for the multiple haplotype comparisons within the class I region have been submitted to DDBJ/EMBL/GenBank under accession nos. AF029061, AF029062, and AB031005-AB031010. Additional sequence data have been submitted to the DDBJ data library under accession nos. AB031005-AB03101 and AF029061-AF029062.] PMID- 11042156 TI - A genetic linkage map of the apicomplexan protozoan parasite Eimeria tenella. AB - Apicomplexan protozoan parasites have complex life cycles that involve phases of asexual and sexual reproduction. Some genera have intermediate insect hosts, for example, Plasmodium spp. (the cause of malaria), but related genera such as Eimeria spp. (causative agents of coccidiosis in poultry) have a direct life cycle occurring in only a single host. Mechanisms that regulate the life cycles of apicomplexan parasites are unknown, but the intracellular growth of avian Eimeria spp. is easily shortened by serial selection for the first parasites to complete the transition from asexual to sexual reproduction (to yield so-called precocious lines). To investigate the genetic basis of such an abbreviated life cycle, we have used the species E. tenella and analyzed the inheritance of 443 polymorphic DNA markers in 22 recombinant cloned progeny derived from a cross between parents that had selectable phenotypes of precocious development or resistance to an anticoccidial drug. The markers were placed in 16 linkage groups (which defined 12 chromosomes) and a further 57 unlinked groups. Two linkage groups showed an association (P =.0105) with the traits of precocious development or drug-resistance and were mapped to chromosome 2 (ca 1.2 Mbp) and chromosome 1 (ca 1.0 Mbp), respectively. The map provides a framework for further studies on the identification of genetic loci implicated in the regulation of the life cycle of an important protozoan parasite and a representative of a major taxonomic group. [A table with the segregation data is available as an online supplement at http://www.genome.org.] PMID- 11042157 TI - Analyzing DNA strand compositional asymmetry to identify candidate replication origins of Borrelia burgdorferi linear and circular plasmids. AB - The Lyme disease agent Borrelia burgdorferi has a genome composed of a linear chromosome and a series of linear and circular plasmids. We previously mapped the oriC of the linear chromosome to the center of the molecule, where a pronounced switch in CG skew occurs. In this study, we analyzed B. burgdorferi plasmid sequences for AT and CG skew in an effort to similarly identify plasmid replication origins. Cumulative skew diagrams of the plasmids suggested that they, like the linear chromosome, replicate bidirectionally from an internal origin. The B. burgdorferi linear chromosome contains homologs to partitioning protein genes soj and spoOJ, which are closely linked to oriC at the minimum cumulative skew point of the 1-Mb molecule. A soj/parA homolog also maps to cumulative skew minima of the B. burgdorferi linear and circular plasmids, further suggesting that these regions contain the replication origin. The heterogeneity in these genes and in the nucleotide sequences of the putative origin regions could account for the mutual compatibility of the multiple DNA elements in B. burgdorferi. PMID- 11042158 TI - Construction and transposition of a 100-kilobase extended P element in Drosophila. AB - We have used P element deletion derivatives at defined locations in the Drosophila genome to construct a 100-kb extended P element more than twice the size of any previously available. We demonstrate that this prototypical extended P element is capable of transposition to new sites in the genome. The structural and functional integrity of a transposed extended P element was confirmed using molecular, genetic, and cytogenetic criteria. This is the first method shown to be capable of producing large, unlinked transpositional duplications in Drosophila. The ability to produce functional transposable elements from half elements is novel and has many potential applications for the functional analysis of complex genomes. PMID- 11042159 TI - Normalization and subtraction of cap-trapper-selected cDNAs to prepare full length cDNA libraries for rapid discovery of new genes. AB - In the effort to prepare the mouse full-length cDNA encyclopedia, we previously developed several techniques to prepare and select full-length cDNAs. To increase the number of different cDNAs, we introduce here a strategy to prepare normalized and subtracted cDNA libraries in a single step. The method is based on hybridization of the first-strand, full-length cDNA with several RNA drivers, including starting mRNA as the normalizing driver and run-off transcripts from minilibraries containing highly expressed genes, rearrayed clones, and previously sequenced cDNAs as subtracting drivers. Our method keeps the proportion of full length cDNAs in the subtracted/normalized library high. Moreover, our method dramatically enhances the discovery of new genes as compared to results obtained by using standard, full-length cDNA libraries. This procedure can be extended to the preparation of full-length cDNA encyclopedias from other organisms. PMID- 11042160 TI - An assessment of gene prediction accuracy in large DNA sequences. AB - One of the first useful products from the human genome will be a set of predicted genes. Besides its intrinsic scientific interest, the accuracy and completeness of this data set is of considerable importance for human health and medicine. Though progress has been made on computational gene identification in terms of both methods and accuracy evaluation measures, most of the sequence sets in which the programs are tested are short genomic sequences, and there is concern that these accuracy measures may not extrapolate well to larger, more challenging data sets. Given the absence of experimentally verified large genomic data sets, we constructed a semiartificial test set comprising a number of short single-gene genomic sequences with randomly generated intergenic regions. This test set, which should still present an easier problem than real human genomic sequence, mimics the approximately 200kb long BACs being sequenced. In our experiments with these longer genomic sequences, the accuracy of GENSCAN, one of the most accurate ab initio gene prediction programs, dropped significantly, although its sensitivity remained high. Conversely, the accuracy of similarity-based programs, such as GENEWISE, PROCRUSTES, and BLASTX was not affected significantly by the presence of random intergenic sequence, but depended on the strength of the similarity to the protein homolog. As expected, the accuracy dropped if the models were built using more distant homologs, and we were able to quantitatively estimate this decline. However, the specificities of these techniques are still rather good even when the similarity is weak, which is a desirable characteristic for driving expensive follow-up experiments. Our experiments suggest that though gene prediction will improve with every new protein that is discovered and through improvements in the current set of tools, we still have a long way to go before we can decipher the precise exonic structure of every gene in the human genome using purely computational methodology. PMID- 11042161 TI - Links from genome proteins to known 3-D structures. AB - We describe a genome annotation service provided by the Entrez browser, http://www.ncbi.nlm.nih.gov/entrez. All protein products identified in fully sequenced microbial genomes have been compared with proteins with known 3-D structure by use of the BLAST sequence comparison algorithm. For the approximately 20% of genome proteins in which unambiguous sequence similarity is detected, Entrez provides a link from the gene product to its predicted structure. The service uses the Cn3D molecular graphics viewer to present a 3-D view of the known structure, together with an alignment display mapping conserved residues from the genome protein onto the known structure. Using an example from Aeropyrum pernix, we illustrate how mapping to a 3-D structure can confirm predictions of biological function. PMID- 11042162 TI - Functional properties of the active core of human cystathionine beta-synthase crystals. AB - Human cystathionine beta-synthase is a pyridoxal 5'-phosphate enzyme containing a heme binding domain and an S-adenosyl-l-methionine regulatory site. We have investigated by single crystal microspectrophotometry the functional properties of a mutant lacking the S-adenosylmethionine binding domain. Polarized absorption spectra indicate that oxidized and reduced hemes are reversibly formed. Exposure of the reduced form of enzyme crystals to carbon monoxide led to the complete release of the heme moiety. This process, which takes place reversibly and without apparent crystal damage, facilitates the preparation of a heme-free human enzyme. The heme-free enzyme crystals exhibited polarized absorption spectra typical of a pyridoxal 5'-phosphate-dependent protein. The exposure of these crystals to increasing concentrations of the natural substrate l-serine readily led to the formation of the key catalytic intermediate alpha-aminoacrylate. The dissociation constant of l-serine was found to be 6 mm, close to that determined in solution. The amount of the alpha-aminoacrylate Schiff base formed in the presence of l-serine was pH independent between 6 and 9. However, the rate of the disappearance of the alpha-aminoacrylate, likely forming pyruvate and ammonia, was found to increase at pH values higher than 8. Finally, in the presence of homocysteine the alpha-aminoacrylate-enzyme absorption band readily disappears with the concomitant formation of the absorption band of the internal aldimine, indicating that cystathionine beta-synthase crystals catalyze both beta elimination and beta-replacement reactions. Taken together, these findings demonstrate that the heme moiety is not directly involved in the condensation reaction catalyzed by cystathionine beta-synthase. PMID- 11042163 TI - Selective role of G protein gamma subunits in receptor interaction. AB - Receptor stimulation of nucleotide exchange in a heterotrimeric G protein (alphabetagamma) is the primary event-modulating signaling by G proteins. The molecular mechanisms at the basis of this event and the role of the G protein subunits, especially the betagamma complex, in receptor activation are unclear. In a reconstituted system, a purified muscarinic receptor, M2, activates G protein heterotrimers alphai2beta1gamma5 and alphai2beta1gamma7 with equal efficacy. However, when the alpha subunit type is substituted with alphao, alphaobeta1gamma7 shows a 100% increase in M2-stimulated GTP hydrolysis compared with alphaobeta1gamma5. Using a sensitive assay based on betagamma complex stimulation of phospholipase C activity, we show that both beta1gamma5 and beta1gamma7 form heterotrimers equally well with alphao and alphai. These results indicate that the gamma subunit interaction with a receptor is critical for modulating nucleotide exchange and is influenced by the subunit-type composition of the heterotrimer. PMID- 11042164 TI - Cd19-dependent activation of Akt kinase in B-lymphocytes. AB - CD19 is rapidly phosphorylated upon B-cell antigen receptor (BCR) cross-linking, leading to the recruitment of downstream signaling intermediates. A prominent feature of CD19 signaling is the binding and activation of phosphoinositide 3 kinase (P13K), which accounts for the majority of PI3K activity induced by BCR ligation. Recent findings have implicated activation of the serine/threonine kinase Akt as imparting survival signals in a PI3K-dependent fashion. Using CD19 deficient B-lymphoma cells and mouse splenic B-cells, we show that CD19 is necessary for efficient activation of Akt following cross-linking of surface immunoglobulin or Igbeta. In the absence of CD19, Akt kinase activity is reduced and transient. In addition, coligation of CD19 with surface immunoglobulin leads to augmented Akt activity in a dose-dependent manner. Thus, CD19 is a key regulator of Akt activity in B-cells; as such it may contribute to pre-BCR or BCR mediated cell survival in vivo. PMID- 11042165 TI - Mutational analysis of protein substrate presentation in the post-translational attachment of biotin to biotin domains. AB - Biotinylation in vivo is an extremely selective post-translational event where the enzyme biotin protein ligase (BPL) catalyzes the covalent attachment of biotin to one specific and conserved lysine residue of biotin-dependent enzymes. The biotin-accepting lysine, present in a conserved Met-Lys-Met motif, resides in a structured domain that functions as the BPL substrate. We have employed phage display coupled with a genetic selection to identify determinants of the biotin domain (yPC-104) of yeast pyruvate carboxylase 1 (residues 1075-1178) required for interaction with BPL. Mutants isolated using this strategy were analyzed by in vivo biotinylation assays performed at both 30 degrees C and 37 degrees C. The temperature-sensitive substrates were reasoned to have structural mutations, leading to compromised conformations at the higher temperature. This interpretation was supplemented by molecular modeling of yPC-104, since these mutants mapped to residues involved in defining the structure of the biotin domain. In contrast, substitution of the Met residue N-terminal to the target lysine with either Val or Thr produced mutations that were temperature insensitive in the in vivo assay. Furthermore, these two mutant proteins and wild type yPC-104 showed identical susceptibility to trypsin, consistent with these substitutions having no structural effect. Kinetic analysis of enzymatic biotinylation using purified Met --> Thr/Val mutant proteins with both yeast and Escherichia coli BPLs revealed that these substitutions had a strong effect upon K(m) values but not k(cat). The Met --> Thr mutant was a poor substrate for both BPLs, whereas the Met --> Val substitution was a poor substrate for bacterial BPL but had only a 2-fold lower affinity for yeast BPL than the wild-type peptide. Our data suggest that substitution of Thr or Val for the Met N-terminal of the biotinyl-Lys results in mutants specifically compromised in their interaction with BPL. PMID- 11042166 TI - Identification and characterization of three novel beta 1,3-N acetylglucosaminyltransferases structurally related to the beta 1,3 galactosyltransferase family. AB - We have isolated three types of cDNAs encoding novel beta1,3-N acetylglucosaminyltransferases (designated beta3Gn-T2, -T3, and -T4) from human gastric mucosa and the neuroblastoma cell line SK-N-MC. These enzymes are predicted to be type 2 transmembrane proteins of 397, 372, and 378 amino acids, respectively. They share motifs conserved among members of the beta1,3 galactosyltransferase family and a beta1,3-N-acetylglucosaminyltransferase (designated beta3Gn-T1), but show no structural similarity to another type of beta1,3-N-acetylglucosaminyltransferase (iGnT). Each of the enzymes expressed by insect cells as a secreted protein fused to the FLAG peptide showed beta1,3-N acetylglucosaminyltransferase activity for type 2 oligosaccharides but not beta1,3-galactosyltransferase activity. These enzymes exhibited different substrate specificity. Transfection of Namalwa KJM-1 cells with beta3Gn-T2, -T3, or -T4 cDNA led to an increase in poly-N-acetyllactosamines recognized by an anti i-antigen antibody or specific lectins. The expression profiles of these beta3Gn Ts were different among 35 human tissues. beta3Gn-T2 was ubiquitously expressed, whereas expression of beta3Gn-T3 and -T4 was relatively restricted. beta3Gn-T3 was expressed in colon, jejunum, stomach, esophagus, placenta, and trachea. beta3Gn-T4 was mainly expressed in brain. These results have revealed that several beta1,3-N-acetylglucosaminyltransferases form a family with structural similarity to the beta1,3-galactosyltransferase family. Considering the differences in substrate specificity and distribution, each beta1,3-N acetylglucosaminyltransferase may play different roles. PMID- 11042167 TI - Disulfide-dependent folding and export of Escherichia coli DsbC. AB - DsbC, a member of the Dsb family in the periplasm of Gram-negative bacteria, is not only a disulfide isomerase but also a chaperone. Five DsbC mutants with Cys in the active site sequence of Cys(98)-Gly-Tyr-Cys(101) and the nonactive site disulfide Cys(141)-Cys(163) replaced by Ser have been studied. The results show that the active site Cys residues are necessary for enzyme activities but not required for chaperone activity, while the lack of the nonactive site disulfide results in a decreased chaperone activity in assisting the reactivation of denatured d-glyceraldehyde-3-phosphate dehydrogenase but has no effect on enzyme activities. Wild-type DsbC was overexpressed and correctly processed as a soluble periplasmic protein. Mutation in one of these Cys residues results in aggregation or extracellular/membrane locations, but does not affect the proper processing. DsbC mutated in either Cys residue of nonactive site disulfide shows higher sensitivity to unfolding by guanidine hydrochloride and slower refolding compared with wild-type DsbC and the active site Cys mutants. The above results provide experimental evidence for structural role of the nonactive site disulfide in folding and biological activities of DsbC. PMID- 11042168 TI - Selective interaction of AGS3 with G-proteins and the influence of AGS3 on the activation state of G-proteins. AB - AGS3 (activator of G-protein signaling 3) was isolated in a yeast-based functional screen for receptor-independent activators of heterotrimeric G proteins. As an initial approach to define the role of AGS3 in mammalian signal processing, we defined the AGS3 subdomains involved in G-protein interaction, its selectivity for G-proteins, and its influence on the activation state of G protein. Immunoblot analysis with AGS3 antisera indicated expression in rat brain, the neuronal-like cell lines PC12 and NG108-15, as well as the smooth muscle cell line DDT(1)-MF2. Immunofluorescence studies and confocal imaging indicated that AGS3 was predominantly cytoplasmic and enriched in microdomains of the cell. AGS3 coimmunoprecipitated with Galpha(i3) from cell and tissue lysates, indicating that a subpopulation of AGS3 and Galpha(i) exist as a complex in the cell. The coimmunoprecipitation of AGS3 and Galpha(i) was dependent upon the conformation of Galpha(i3) (GDP GTPgammaS (guanosine 5'-3-O-(thio)triphosphate)). The regions of AGS3 that bound Galpha(i) were localized to four amino acid repeats (G-protein regulatory motif (GPR)) in the carboxyl terminus (Pro(463) Ser(650)), each of which were capable of binding Galpha(i). AGS3-GPR domains selectively interacted with Galpha(i) in tissue and cell lysates and with purified Galpha(i)/Galpha(t). Subsequent experiments with purified Galpha(i2) and Galpha(i3) indicated that the carboxyl-terminal region containing the four GPR motifs actually bound more than one Galpha(i) subunit at the same time. The AGS3 GPR domains effectively competed with Gbetagamma for binding to Galpha(t(GDP)) and blocked GTPgammaS binding to Galpha(i1). AGS3 and related proteins provide unexpected mechanisms for coordination of G-protein signaling pathways. PMID- 11042169 TI - Up-regulation of endothelial nitric-oxide synthase promoter by the phosphatidylinositol 3-kinase gamma /Janus kinase 2/MEK-1-dependent pathway. AB - Our recent study indicates that lysophosphatidylcholine (LPC) enhances Sp1 binding and Sp1-dependent endothelial nitric oxide synthase (eNOS) promoter activity via the mitogen-activated protein kinase/extracellular signal-regulated kinase kinase 1 (MEK-1) signaling pathway (Cieslik, K., Lee, C.-M., Tang, J.-L., and Wu, K. K. (1999) J. Biol. Chem. 274, 34669-34675). To identify upstream signaling molecules, we transfected human endothelial cells with dominant negative and active mutants of Ras and evaluated their effects on eNOS promoter activity. Neither mutant altered the basal or LPC-induced eNOS promoter function. By contrast, a dominant negative mutant of phosphatidylinositol 3-kinase gamma (PI-3Kgamma) blocked the promoter activity induced by LPC. Wortmannin and LY 294002 had a similar effect. AG-490, a selective inhibitor of Janus kinase 2 (Jak2), also reduced the LPC-induced Sp1 binding and eNOS promoter activity to the basal level. LPC induced Jak2 phosphorylation, which was abolished by LY 294002 and the dominant negative mutant of PI-3Kgamma. LY 294002 and AG-490 abrogated MEK-1 phosphorylation induced by LPC but had no effect on Raf-1. These results indicate that PI-3Kgamma and Jak2 are essential for LPC-induced eNOS promoter activity. This signaling pathway was sensitive to pertussis toxin, suggesting the involvement of a G(i) protein in PI-3Kgamma activation. These results indicate that LPC enhances Sp1-dependent eNOS promoter activity by a pertussis toxin-sensitive, Ras-independent novel pathway, PI-3Kgamma/Jak2/MEK 1/ERK1/2. PMID- 11042170 TI - Domain-dependent function of the rasGAP-binding protein p62Dok in cell signaling. AB - p62Dok, the rasGAP-binding protein, is a common target of protein-tyrosine kinases. It is one of the major tyrosine-phosphorylated molecules in v-Src transformed cells. Dok consists of an amino-terminal Pleckstrin homology domain, a putative phosphotyrosine binding domain, and a carboxyl-terminal tail containing multiple tyrosine phosphorylation sites. The importance and function of these sequences in Dok signaling remain largely unknown. We have demonstrated here that the expression of Dok can inhibit cellular transformation by the Src tyrosine kinase. Both the phosphotyrosine binding domain and the carboxyl terminal tail of Dok (in particular residues 336-363) are necessary for such activity. Using a combinatorial peptide library approach, we have shown that the Dok phosphotyrosine binding domain binds phosphopeptides with the consensus motif of Y/MXXNXL-phosphotyrosine. Furthermore, Dok can homodimerize through its phosphotyrosine binding domain and Tyr(146) at the amino-terminal region. Mutations of this domain or Tyr(146) that block homodimerization significantly reduce the ability of Dok to inhibit Src transformation. Our results suggest that Dok oligomerization through its multiple domains plays a critical role in Dok signaling in response to tyrosine kinase activation. PMID- 11042171 TI - Molecular cloning and characterization of a novel regulator of G-protein signaling from mouse hematopoietic stem cells. AB - A novel regulator of G-protein signaling (RGS) has been isolated from a highly purified population of mouse long-term hematopoietic stem cells, and designated RGS18. It has 234 amino acids consisting of a central RGS box and short divergent NH(2) and COOH termini. The calculated molecular weight of RGS18 is 27,610 and the isoelectric point is 8.63. Mouse RGS18 is expressed from a single gene and shows tissue specific distribution. It is most highly expressed in bone marrow followed by fetal liver, spleen, and then lung. In bone marrow, RGS18 level is highest in long-term and short-term hematopoietic stem cells, and is decreased as they differentiate into more committed multiple progenitors. The human RGS18 ortholog has a tissue-specific expression pattern similar to that of mouse RGS18. Purified RGS18 interacts with the alpha subunit of both G(i) and G(q) subfamilies. The results of in vitro GTPase single-turnover assays using Galpha(i) indicated that RGS18 accelerates the intrinsic GTPase activity of Galpha(i). Transient overexpression of RGS18 attenuated inositol phosphates production via angiotensin receptor and transcriptional activation through cAMP responsive element via M1 muscarinic receptor. This suggests RGS18 can act on G(q)-mediated signaling pathways in vivo. PMID- 11042172 TI - Hoxa-9 represses transforming growth factor-beta-induced osteopontin gene transcription. AB - Smad2 and Smad3 are downstream transforming growth factor-beta (TGF-beta) signaling molecules. Upon phosphorylation by its type I receptor, Smad2 or Smad3 forms a complex with Smad4 and translocates to the nucleus where the complex activates target gene transcription. In the present study, we report that Smad3 binds directly to the osteopontin (OPN) promoter and that Smad4 interacts with the Hox protein and displaces it from its cognate DNA binding site in response to TGF-beta stimulation. In gel shift assays, the glutathione S-transferase-Smad3 fusion protein was found to bind to a 50-base pair DNA element (-179 to -229) from the OPN promoter. Also, we found that both Hoxc-8 and Hoxa-9 bound to a Hox binding site adjacent to Smad3 binding sequence. Interestingly, Smad4, the common partner for both bone morphogenic protein and TGF-beta signaling pathways, inhibited the binding of Hox protein to DNA. FLAG-tagged Smad4 coimmunoprecipitated with HA-tagged Hoxa-9 from cotransfected COS-1 cells, demonstrating an interaction between Smad4 and Hoxa-9. Transfection studies showed that Hoxa-9 is a strong transcriptional repressor; it suppresses the transcription of the luciferase reporter gene driven by a 124-base pair OPN promoter fragment containing both Smad3 and Hox binding sites. Taken together, these data demonstrate a unique TGF-beta-induced transcription mechanism. Smad3 and Smad4 exhibit different functions in activation of OPN transcription. Smad3 binds directly to the OPN promoter as a sequence-specific activator, and Smad4 displaces the transcription repressor, Hoxa-9, by formation of Smad4/Hox complex as part of the transcription mechanism in response to TGF-beta stimulation. PMID- 11042173 TI - Proteomics characterization of abundant Golgi membrane proteins. AB - A mass spectrometric analysis of proteins partitioning into Triton X-114 from purified hepatic Golgi apparatus (84% purity by morphometry, 122-fold enrichment over the homogenate for the Golgi marker galactosyl transferase) led to the unambiguous identification of 81 proteins including a novel Golgi-associated protein of 34 kDa (GPP34). The membrane protein complement was resolved by SDS polyacrylamide gel electrophoresis and subjected to a hierarchical approach using delayed extraction matrix-assisted laser desorption ionization mass spectrometry characterization by peptide mass fingerprinting, tandem mass spectrometry to generate sequence tags, and Edman sequencing of proteins. Major membrane proteins corresponded to known Golgi residents, a Golgi lectin, anterograde cargo, and an abundance of trafficking proteins including KDEL receptors, p24 family members, SNAREs, Rabs, a single ARF-guanine nucleotide exchange factor, and two SCAMPs. Analytical fractionation and gold immunolabeling of proteins in the purified Golgi fraction were used to assess the intra-Golgi and total cellular distribution of GPP34, two SNAREs, SCAMPs, and the trafficking proteins GBF1, BAP31, and alpha(2)P24 identified by the proteomics approach as well as the endoplasmic reticulum contaminant calnexin. Although GPP34 has never previously been identified as a protein, the localization of GPP34 to the Golgi complex, the conservation of GPP34 from yeast to humans, and the cytosolically exposed location of GPP34 predict a role for a novel coat protein in Golgi trafficking. PMID- 11042174 TI - Unique oxidative mechanisms for the reactive nitrogen oxide species, nitroxyl anion. AB - The nitroxyl anion (NO-) is a highly reactive molecule that may be involved in pathophysiological actions associated with increased formation of reactive nitrogen oxide species. Angeli's salt (Na2N2O3; AS) is a NO- donor that has been shown to exert marked cytotoxicity. However, its decomposition intermediates have not been well characterized. In this study, the chemical reactivity of AS was examined and compared with that of peroxynitrite (ONOO-) and NO/N2O3. Under aerobic conditions, AS and ONOO- exhibited similar and considerably higher affinities for dihydrorhodamine (DHR) than NO/N2O3. Quenching of DHR oxidation by azide and nitrosation of diaminonaphthalene were exclusively observed with NO/N2O3. Additional comparison of ONOO- and AS chemistry demonstrated that ONOO- was a far more potent one-electron oxidant and nitrating agent of hydroxyphenylacetic acid than was AS. However, AS was more effective at hydroxylating benzoic acid than was ONOO-. Taken together, these data indicate that neither NO/N2O3 nor ONOO- is an intermediate of AS decomposition. Evaluation of the stoichiometry of AS decomposition and O2 consumption revealed a 1:1 molar ratio. Indeed, oxidation of DHR mediated by AS proved to be oxygen-dependent. Analysis of the end products of AS decomposition demonstrated formation of NO2- and NO3- in approximately stoichiometric ratios. Several mechanisms are proposed for O2 adduct formation followed by decomposition to NO3- or by oxidation of an HN2O3- molecule to form NO2-. Given that the cytotoxicity of AS is far greater than that of either NO/N2O3 or NO + O2, this study provides important new insights into the implications of the potential endogenous formation of NO- under inflammatory conditions in vivo. PMID- 11042175 TI - Identification and characterization of the carboxyl-terminal region of rat dentin sialoprotein. AB - Two acidic proteins, dentin sialoprotein (DSP) and dentin phosphoprotein (DPP), are present in the extracellular matrix of dentin but not in bone. These two proteins are expressed in odontoblasts and preameloblasts as a single cDNA transcript coding a large precursor protein termed dentin sialophosphoprotein (DSPP). DSPP is specifically cleaved into two unique proteins, DSP and DPP. However, the cleavage site(s) of DSPP and the mechanisms for regulating the cleavages are unknown. To identify the specific site(s) of DSPP that are cleaved when the initial translation product is converted to DSP and DPP, we performed a detailed analysis (Edman degradation and mass spectrometry) on selected tryptic peptides of a size originating from the COOH-terminal region of rat DSP. After cleavage with trypsin, the DSP fragments were separated by a two-dimensional method (size-exclusion chromatography followed by reversed phase high performance liquid chromatography). We characterized 13 peptides from various regions of DSP. The analyses showed that peptide Ile(409)-Tyr(421) was the major COOH-terminal fragment, ending at Tyr(421) only 9 residues from the NH(2) terminus of DPP. Peptide Gln(385)-His(406) represented a second, minor COOH-terminal peptide that terminated at His(406). Both of these residues are well beyond the COOH terminus predicted previously by two independent studies estimating that rat DSP contained 360-370 amino acids. Careful studies on two peptides showed that, among 9 potential casein kinase II phosphorylation sites, 2 serines were phosphorylated. We found that rat DSP was heterogeneous with respect to phosphorylation, because this same peptide sequence eluted in two discrete peaks, one with 2 phosphoserines and the other having 1. The finding that 3 lysines just preceding the COOH termini were modified by a 43-Da substituent (possibly a carbamoyl substituent) suggests that the lysines in this region were particularly susceptible to attachment of this substituent. PMID- 11042176 TI - Site-directed mutagenesis of human ceruloplasmin:. production of a proteolytically stable protein and structure-activity relationships of type 1 sites. AB - A fully active recombinant human ceruloplasmin was obtained, and it was mutated to produce a ceruloplasmin stable to proteolysis. The stable ceruloplasmin was further mutated to perturb the environment of copper at the type 1 copper sites in two different domains. The wild type and the mutated ceruloplasmin were produced in the yeast Pichia pastoris and characterized. The mutations R481A, R701A, and K887A were at the proteolytic sites, did not alter the enzymatic activity, and were all necessary to protect ceruloplasmin from degradation. The mutation L329M was at the tricoordinate type 1 site of the domain 2 and was ineffective to induce modifications of the spectroscopic and catalytic properties of ceruloplasmin, supporting the hypothesis that this site is reduced and locked in a rigid frame. In contrast the mutation C1021S at the type 1 site of domain 6 substantially altered the molecular properties of the protein, leaving a small fraction endowed with oxidase activity. This result, while indicating the importance of this site in stabilizing the overall protein structure, suggests that another type 1 site is competent for dioxygen reduction. During the expression of ceruloplasmin, the yeast maintained a high level of Fet3 that was released from membranes of yeast not harboring the ceruloplasmin gene. This indicates that expression of ceruloplasmin induces a state of iron deficiency in yeast because the ferric iron produced in the medium by its ferroxidase activity is not available for the uptake. PMID- 11042177 TI - Plant initiation factor 3 subunit composition resembles mammalian initiation factor 3 and has a novel subunit. AB - Eukaryotic initiation factor 3 (eIF3) is a multisubunit complex that is required for binding of mRNA to 40 S ribosomal subunits, stabilization of ternary complex binding to 40 S subunits, and dissociation of 40 and 60 S subunits. These functions and the complex nature of eIF3 suggest multiple interactions with many components of the translational machinery. Recently, the subunits of mammalian and Saccharomyces cerevisiae eIF3 were identified, and substantial differences in the subunit composition of mammalian and S. cerevisiae were observed. Mammalian eIF3 consists of 11 nonidentical subunits, whereas S. cerevisiae eIF3 consists of up to eight nonidentical subunits. Only five of the subunits of mammalian and S. cerevisiae are shared in common, and these five subunits comprise a "core" complex in S. cerevisiae. eIF3 from wheat consists of at least 10 subunits, but their relationship to either the mammalian or S. cerevisiae eIF3 subunits is unknown. Peptide sequences derived from purified wheat eIF3 subunits were used to correlate each subunit with mammalian and/or S. cerevisiae subunits. The peptide sequences were also used to identify Arabidopsis thaliana cDNAs for each of the eIF3 subunits. We report seven new cDNAs for A. thaliana eIF3 subunits. A. thaliana eIF3 was purified and characterized to confirm that the subunit composition and activity of wheat and A. thaliana eIF3 were similar. We report that plant eIF3 closely resembles the subunit composition of mammalian eIF3, having 10 out of 11 subunits in common. Further, we find a novel subunit in the plant eIF3 complex not present in either mammalian or S. cerevisiae eIF3. These results suggest that plant and mammalian eIF3 evolved similarly, whereas S. cerevisiae has diverged. PMID- 11042178 TI - Guard cell inward K+ channel activity in arabidopsis involves expression of the twin channel subunits KAT1 and KAT2. AB - Stomatal opening, which controls gas exchanges between plants and the atmosphere, results from an increase in turgor of the two guard cells that surround the pore of the stoma. KAT1 was the only inward K(+) channel shown to be expressed in Arabidopsis guard cells, where it was proposed to mediate a K(+) influx that enables stomatal opening. We report that another Arabidopsis K(+) channel, KAT2, is expressed in guard cells. More than KAT1, KAT2 displays functional features resembling those of native inward K(+) channels in guard cells. Coexpression in Xenopus oocytes and two-hybrid experiments indicated that KAT1 and KAT2 can form heteromultimeric channels. The data indicate that KAT2 plays a crucial role in the stomatal opening machinery. PMID- 11042179 TI - Growth hormone receptor ubiquitination coincides with recruitment to clathrin coated membrane domains. AB - Endocytosis of the growth hormone receptor (GHR) depends on a functional ubiquitin conjugation system. A 10-amino acid residue motif within the GHR cytosolic tail (the ubiquitin-dependent endocytosis motif) is involved in both GHR ubiquitination and endocytosis. As shown previously, ubiquitination of the receptor itself is not required. In this paper ubiquitination of the GHR was used as a tool to address the question of at which stage the ubiquitin conjugation system acts in the process of GHR endocytosis. If potassium depletion was used to interfere with early stages of coated pit formation, both GHR endocytosis and ubiquitination were inhibited. Treatment of cells with methyl-beta-cyclodextrin inhibited endocytosis at the stage of coated vesicle formation. Growth hormone addition to methyl-beta-cyclodextrin-treated cells resulted in an accumulation of ubiquitinated GHR at the cell surface. Using immunoelectron microscopy, the GHR was localized in flattened clathrin-coated membranes. In addition, when clathrin mediated endocytosis was inhibited in HeLa cells expressing a temperature sensitive dynamin mutant, ubiquitinated GHR accumulated at the cell surface. Together, these data show that the GHR is ubiquitinated at the plasma membrane, before endocytosis occurs, and indicate that the resident time of the GHR at the cell surface is regulated by the ubiquitin conjugation system together with the endocytic machinery. PMID- 11042180 TI - The Cdc42p GTPase and its regulators Nrf1p and Scd1p are involved in endocytic trafficking in the fission yeast Schizosaccharomyces pombe. AB - Nrf1p was first identified in a screen for negative regulators of the Cdc42p GTPase. Overexpression of Nrf1p resulted in dose-dependent lethality, with cells exhibiting an ellipsoidal morphology and abnormal vacuolar phenotypes including an increase in vacuolar fusion. Green fluorescent protein (GFP)-Cdc42p and GFP Nrf1p colocalized to vacuolar membranes and GFP-Nrf1p vacuolar localization depended on Scd1p, the Schizosaccharomyces pombe homolog of the Cdc24p guanine nucleotide exchange factor. In this study, site-directed mutagenesis was conducted on Nrf1p to determine its functional domains. Mutations in the three putative transmembrane domains resulted in mislocalization of GFP-Nrf1p and an inability to induce lethality, suggesting a loss of function. Mutations in the second extramembranous loop of Nrf1p also resulted in a loss of function and altered the ability of GFP-Nrf1p to localize to vacuolar membranes. Analysis of Deltanrf1 and Deltascd1 mutants revealed defects in endocytosis. In addition, overexpression of constitutively active Cdc42(G12V)p resulted in an increase in endocytosis and an ability to rescue the endocytic defects in Deltanrf1 and Deltascd1 cells. These data are consistent with Nrf1p and Scd1p being necessary for efficient endocytosis, possibly through the regulation of Cdc42p. PMID- 11042181 TI - Molecular characterization and developmental expression of NORPEG, a novel gene induced by retinoic acid. AB - We have characterized NORPEG, a novel gene from human retinal pigment epithelial cells (ARPE-19), in which its expression is induced by all-trans-retinoic acid. Two transcripts ( approximately 3 and approximately 5 kilobases in size) have been detected for this gene, which is localized to chromosome band 5p13.2-13.3. Placenta and testis showed the highest level of expression among various human tissues tested. Six ankyrin repeats and a long coiled-coil domain are present in the predicted sequence of the NORPEG protein, which contains 980 amino acid residues. This approximately 110-kDa protein was transiently expressed in COS-7 cells as a FLAG fusion protein and immunolocalized to the cytoplasm. Confocal microscopic analysis of the NORPEG protein in ARPE-19 cells showed threadlike projections in the cytoplasm reminiscent of the cytoskeleton. Consistent with this localization, the expressed NORPEG protein showed resistance to solubilization by Triton X-100 and KCl. An ortholog of NORPEG characterized from mouse encoded a protein that showed 91% sequence similarity to the human NORPEG protein. The expression of Norpeg mRNA was detected in mouse embryo at embryonic day 9.5 by in situ hybridization, and the expression appears to be developmentally regulated. In adult mouse, the highest level of expression was detected in the seminiferous tubules of testis. PMID- 11042182 TI - A novel mechanism of cooperation between c-Kit and erythropoietin receptor. Stem cell factor induces the expression of Stat5 and erythropoietin receptor, resulting in efficient proliferation and survival by erythropoietin. AB - Optimal production of red cells in vivo requires collaboration between c-Kit, erythropoietin receptor (Epo-R), and GATA-1. However, the mechanism(s) of collaboration remain unclear. Utilizing an embryonic stem cell-derived erythroid progenitor cell line from mice deficient in GATA-1, we have examined the role of c-Kit and Epo-R in erythroid cell proliferation, survival, and differentiation. In the absence of GATA-1, we demonstrate an essential role for c-Kit in survival and proliferation of erythroid progenitors via the regulation of Bcl-2 expression. In addition, we demonstrate that Epo-R and Stat5 are regulated by a second, novel mechanism. We demonstrate that c-Kit stimulation by stem cell factor is essential for the maintenance of Epo-R and Stat5 protein expression, which results in significantly enhanced Bcl-x(L) induction and survival of erythroid progenitors in response to Epo stimulation. Restoration of GATA-1 function results in terminal erythroid maturation and up-regulation of Epo-R and Bcl-x(L) expression, leading also to significantly enhanced survival of terminally differentiating erythroid progenitors in the presence of only Epo. These results demonstrate that c-Kit and Epo-R have unique role(s) during distinct phases of erythroid maturation, and both stem cell factor and Epo contribute to the regulation of the Epo-R-Stat5-Bcl-x(L) pathway to ensure optimal survival, proliferation, and differentiation of erythroid progenitors. PMID- 11042183 TI - Lysophosphatidic acid-induced mitogenesis is regulated by lipid phosphate phosphatases and is Edg-receptor independent. AB - Lysophosphatidic acid (LPA) is an extracellular signaling mediator with a broad range of cellular responses. Three G-protein-coupled receptors (Edg-2, -4, and 7) have been identified as receptors for LPA. In this study, the ectophosphatase lipid phosphate phosphatase 1 (LPP1) has been shown to down-regulate LPA-mediated mitogenesis. Furthermore, using degradation-resistant phosphonate analogs of LPA and stereoselective agonists of the Edg receptors we have demonstrated that the mitogenic and platelet aggregation responses to LPA are independent of Edg-2, -4, and -7. Specifically, we found that LPA degradation is insufficient to account for the decrease in LPA potency in mitogenic assays, and the stereoselectivity observed at the Edg receptors is not reflected in mitogenesis. Additionally, RH7777 cells, which are devoid of Edg-2, -4, and -7 receptor mRNA, have a mitogenic response to LPA and LPA analogs. Finally, we have determined that the ligand selectivity of the platelet aggregation response is consistent with that of mitogenesis, but not with Edg-2, -4, and -7. PMID- 11042184 TI - Tissue inhibitor of metalloproteinases-2 (TIMP-2) suppresses TKR-growth factor signaling independent of metalloproteinase inhibition. AB - The tissue inhibitors of metalloproteinases (TIMPs) block matrix metalloproteinase (MMP)-mediated increases in cell proliferation, migration, and invasion that are associated with extracellular matrix (ECM) turnover. Here we demonstrate a direct role for TIMP-2 in regulating tyrosine kinase-type growth factor receptor activation. We show that TIMP-2 suppresses the mitogenic response to tyrosine kinase-type receptor growth factors in a fashion that is independent of MMP inhibition. The TIMP-2 suppression of mitogenesis is reversed by the adenylate cyclase inhibitor SQ22536, and implicates cAMP as the second messenger in these effects. TIMP-2 neither altered the release of transforming growth factor alpha from the cell surface, nor epidermal growth factor (EGF) binding to the cognate receptor, EGFR. TIMP-2 binds to the surface of A549 cells in a specific and saturable fashion (K(d) = 147 pm), that is not competed by the synthetic MMP inhibitor BB-94 and is independent of MT-1-MMP. TIMP-2 induces a decrease in phosphorylation of EGFR and a concomitant reduction in Grb-2 association. TIMP-2 prevents SH2-protein-tyrosine phosphatase-1 (SHP-1) dissociation from immunoprecipitable EGFR complex and a selective increase in total SHP-1 activity. These studies represent a new functional paradigm for TIMP 2 in which TIMP suppresses EGF-mediated mitogenic signaling by short-circuiting EGFR activation. PMID- 11042185 TI - The N-terminal "beta-barrel" domain of 5-lipoxygenase is essential for nuclear membrane translocation. AB - 5-Lipoxygenase is the key enzyme in the formation of leukotrienes, which are potent lipid mediators of asthma pathophysiology. This enzyme translocates to the nuclear envelope in a calcium-dependent manner for leukotriene biosynthesis. Eight green fluorescent protein (GFP)-lipoxygenase constructs, representing the major human and mouse enzymes within this family, were constructed and their cDNAs transfected into human embryonic kidney 293 cells. Of these eight lipoxygenases, only the 5-lipoxygenase was clearly nuclear localized and translocated to the nuclear envelope upon stimulation with the calcium ionophore. The N-terminal "beta -barrel" domain of 5-lipoxygenase, but not the catalytic domain, was necessary and sufficient for nuclear envelope translocation. The GFP N-terminal 5-lipoxygenase domain translocated faster than GFP-5-lipoxygenase. beta-Barrel/catalytic domain chimeras with 12- and 15-lipoxygenase indicated that only the N-terminal domain of 5-lipoxygenase could carry out this translocation function. Mutations of iron atom binding ligands (His550 or deletion of C terminal isoleucine) that disrupt nuclear localization do not alter translocation capacity indicating distinct determinants of nuclear localization and translocation. Moreover, data show that GFP-5-lipoxygenase beta-barrel containing constructs can translocate to the nuclear membrane whether cytoplasmic or nuclear localized. Thus, the predicted beta-barrel domain of 5-lipoxygenase may function like the C2 domain within protein kinase C and cytosolic phospholipase A(2) with unique determinants that direct its localization to the nuclear envelope. PMID- 11042186 TI - Heat shock and oxidative stress-induced exposure of hydrophobic protein domains as common signal in the induction of hsp68. AB - The hypothesis of a common signal for heat shock (HS) and oxidative stress (OS) was analyzed in C6 cells with regard to the induction of heat shock proteins (Hsps). The synthesis rate and level of the strictly inducible Hsp68 was significantly higher after HS (44 degrees C) compared with OS (2 mm H2O2). This difference corresponded to higher and lower activation of the heat shock factor (HSF) by HS and OS, respectively. OS, on the other hand, showed stronger cytotoxicity compared with HS as indicated by drastic lipid peroxidation and inhibition of protein synthesis as well as of mitochondrial and endocytotic activity. Lactic dehydrogenase also revealed stronger inhibition of enzyme activity by OS than by HS as shown in cells and in vitro experiments. Conformational analysis of lactic dehydrogenase by the fluorophore 1 anilinonaphtalene-8-sulfonic acid, however, showed stronger exposure of hydrophobic domains after HS than after OS which correlates positively with the Hsp68 response. Treatment of cells with deoxyspergualin, which exhibits high affinity to Hsps, the putative inhibitors of HSF, strongly increased only OS induced hsp68 expression. In conclusion, the results suggest that exposure of hydrophobic domains of cytosolic proteins represents the common first signal in the multistep activation pathway of HSF. PMID- 11042187 TI - Stable activation of single Ca2+ release-activated Ca2+ channels in divalent cation-free solutions. AB - The regulation of store-operated, calcium-selective channels in the plasma membrane of rat basophilic leukemia cells (RBL-2H3 m1), an immortalized mucosal mast cell line, was studied at the single-channel level with the patch clamp technique by removing divalent cations from both sides of the membrane. The activity of the single channels in excised patches could be modulated by Ca(2+), Mg(2+), and pH. The maximal activation of these channels by divalent cation-free conditions occurred independently of depletion of intracellular Ca(2+) stores, whether in excised patches or in whole cell mode. Yet, a number of points of evidence establish these single-channel openings as amplified store-operated channel events. Specifically, (i) the single channels are exquisitely sensitive to inhibition by intracellular Ca(2+), and (ii) both the store-operated current and the single-channel openings are completely blocked by the capacitative calcium entry blocker, 2-aminoethoxydiphenyl borane. In addition, in Jurkat T cells single-channel openings with lower open probability have been observed in the whole cell mode with intracellular Mg(2+) present (Kerschbaum, H. H., and Cahalan, M. D. (1999) Science 283, 836-839), and in RBL-2H3 m1 cells a current with similar properties is activated by store depletion. PMID- 11042188 TI - The potency and specificity of the interaction between the IA3 inhibitor and its target aspartic proteinase from Saccharomyces cerevisiae. AB - The yeast IA3 polypeptide consists of only 68 residues, and the free inhibitor has little intrinsic secondary structure. IA3 showed subnanomolar potency toward its target, proteinase A from Saccharomyces cerevisiae, and did not inhibit any of a large number of aspartic proteinases with similar sequences/structures from a wide variety of other species. Systematic truncation and mutagenesis of the IA3 polypeptide revealed that the inhibitory activity is located in the N-terminal half of the sequence. Crystal structures of different forms of IA3 complexed with proteinase A showed that residues in the N-terminal half of the IA3 sequence became ordered and formed an almost perfect alpha-helix in the active site of the enzyme. This potent, specific interaction was directed primarily by hydrophobic interactions made by three key features in the inhibitory sequence. Whereas IA3 was cut as a substrate by the nontarget aspartic proteinases, it was not cleaved by proteinase A. The random coil IA3 polypeptide escapes cleavage by being stabilized in a helical conformation upon interaction with the active site of proteinase A. This results, paradoxically, in potent selective inhibition of the target enzyme. PMID- 11042189 TI - Identification and characterization of a novel MAP kinase kinase kinase, MLTK. AB - The MAPK cascades regulate a wide variety of cellular functions, including cell proliferation, differentiation, and stress responses. Here we have identified a novel MAP kinase kinase kinase (MAPKKK), termed MLTK (for MLK-like mitogen activated protein triple kinase), whose expression is increased by activation of the ERK/MAPK pathway. There are two alternatively spliced forms of MLTK, MLTKalpha and MLTKbeta. When overexpressed in cells, both MLTKalpha and MLTKbeta are able to activate the ERK, JNK/SAPK, p38, and ERK5 pathways. Moreover, both MLTKalpha and MLTKbeta are activated in response to osmotic shock with hyperosmolar media through autophosphorylation. Remarkably, expression of MLTKalpha, but not MLTKbeta, in Swiss 3T3 cells results in the disruption of actin stress fibers and dramatic morphological changes. A kinase-dead form of MLTKalpha does not cause these phenomena. Inhibition of the p38 pathway significantly blocks MLTKalpha-induced stress fiber disruption and morphological changes. These results suggest that MLTK is a stress-activated MAPKKK that may be involved in the regulation of actin organization. PMID- 11042190 TI - Analysis of the subsite specificity of rat insulysin using fluorogenic peptide substrates. AB - Recombinant rat insulysin was shown to cleave the internally quenched fluorogenic peptide 2-aminobenzyl-GGFLRKVGQ-ethylenediamine-2,4-dinitrophenol at the R-K bond, exhibiting a K(m) of 13 microm and a V(max) of 2.6 micromol min(-1) mg(-1). Derivatives of this peptide in which the P(2) leucine or the P(2)' valine were replaced with other residues were used to probe the subsite specificity of the enzyme. Varying the P(2) residue produced a 4-fold range in K(m) and a 7-fold range in k(cat). The nature of the P(2) residue had a significant effect on the site of cleavage. Leucine, isoleucine, valine, and aspartate produced cleavage at the R-K bond. Asparagine produced 36% cleavage at the N-R bond and 64% cleavage at the R-K bond, whereas with alanine or serine the A-R and S-R bonds were the major cleavage sites. With tyrosine, phenylalanine, methionine, or histidine representing the varied residue X, cleavages at F-X, X-R, and R-K were seen, whereas with tryptophan equal cleavage occurred at the F-W and W-R bonds. Variable P(2)' residues produce less of a change in both K(m) and k(cat) and have little influence on the cleavage site. Exceptions are phenylalanine, tyrosine, leucine, and isoleucine, which in addition to producing cleavage at the R-K bond, produce significant cleavage at the L-R bond. Alanine and tyrosine were unique in producing cleavage at the F-L bond. Taken together, these data suggest that insulysin specificity is directed toward the amino side of hydrophobic and basic residues and that the enzyme has an extended substrate binding site. PMID- 11042191 TI - Phosphorylation of GRK2 by protein kinase C abolishes its inhibition by calmodulin. AB - G-protein-coupled receptor kinases (GRKs) are important regulators of G-protein coupled receptor function. Two members of this family L, GRK2 and GRK5 L, have been shown to be substrates for protein kinase C (PKC). Whereas PKC-mediated phosphorylation results in inhibition of GRK5, it increases the activity of GRK2 toward its substrates probably through increased affinity for receptor-containing membranes. We show here that this increase in activity may be caused by relieving a tonic inhibition of GRK2 by calmodulin. In vitro, GRK2 was preferentially phosphorylated by PKC isoforms alpha, gamma, and delta. Two-dimensional peptide mapping of PKCalpha-phosphorylated GRK2 showed a single site of phosphorylation, which was identified as serine 29 by HPLC-MS. A S29A mutant of GRK2 was not phosphorylated by PKC in vitro and showed no phorbol ester-stimulated phosphorylation when transfected into human embryonic kidney (HEK)293 cells. Serine 29 is located in the calmodulin-binding region of GRK2, and binding of calmodulin to GRK2 results in inhibition of kinase activity. This inhibition was almost completely abolished in vitro when GRK2 was phosphorylated by PKC. These data suggest that calmodulin may be an inhibitor of GRK2 whose effects can be abolished with PKC-mediated phosphorylation of GRK2. PMID- 11042192 TI - Ca2+-induced phosphoethanolamine transfer to the outer 3-deoxy-D-manno octulosonic acid moiety of Escherichia coli lipopolysaccharide. A novel membrane enzyme dependent upon phosphatidylethanolamine. AB - Certain strains of Escherichia coli and Salmonella contain lipopolysaccharide (LPS) modified with a phosphoethanolamine (pEtN) group at position 7 of the outer 3-deoxy-d-manno-octulosonic acid (Kdo) residue. Using the heptose-deficient E. coli mutant WBB06 (Brabetz, W., Muller-Loennies, S., Holst, O., and Brade, H. (1997) Eur. J. Biochem. 247, 716-724), we now demonstrate that the critical parameter determining the presence or absence of pEtN is the concentration of CaCl(2) in the medium. As judged by mass spectrometry, half the LPS in WBB06, grown on nutrient broth with 5 mm CaCl(2), is derivatized with a pEtN group, whereas LPS from WBB06 grown without supplemental CaCl(2) is not. Membranes from E. coli WBB06 or wild-type W3110 grown on 5-50 mm CaCl(2) contain a novel pEtN transferase that uses the precursor Kdo(2)-[4'-(32)P]lipid IV(A) as an acceptor. Transferase is not present in membranes of E. coli grown with 5 mm MgCl(2), BaCl(2), or ZnCl(2). Hydrolysis of the in vitro reaction product, pEtN-Kdo(2)-[4' (32)P]lipid IV(A), at pH 4.5 shows that the pEtN substituent is located on the outer Kdo moiety. Membranes from an E. coli pss knockout mutant grown on 50 mm CaCl(2), which lack phosphatidylethanolamine, do not contain measurable transferase activity unless exogenous phosphatidylethanolamine is added back to the assay system. The induction of the pEtN transferase by 5-50 mm CaCl(2) suggests possible role(s) in establishing transformation competence or resisting environmental stress, and represents the first example of a regulated covalent modification of the inner core of E. coli LPS. PMID- 11042193 TI - Phospholipase C-gamma 2 couples Bruton's tyrosine kinase to the NF-kappaB signaling pathway in B lymphocytes. AB - Mutations in the gene encoding Bruton's tyrosine kinase (BTK) interfere with B cell proliferation and lead to an X-linked immunodeficiency in mice characterized by reduced B cell numbers. Recent studies have established that BTK transmits signals from the B cell antigen receptor (BCR) to transcription factor NF-kappaB, which in turn reprograms a set of genes required for normal B cell growth. We now demonstrate that induction of NF-kappaB via this pathway requires the intermediate action of the -gamma2 isoform of phospholipase C (PLC-gamma2), a potential phosphorylation substrate of BTK. Specifically, pharmacologic agents that block the action of either PLC-gamma2 or its second messengers prevent BCR induced activation of IkappaB kinase. Moreover, activation of NF-kappaB in response to BCR signaling is completely abolished in B cells deficient for PLC gamma2. Taken together, these findings strongly suggest that PLC-gamma2 functions as an integral component of the BTK/NF-kappaB axis following BCR ligation. Interference with this NF-kappaB cascade may account for some of the B cell defects reported for plc-gamma2(-/-) mice, which develop an X-linked immunodeficiency-like phenotype. PMID- 11042194 TI - Metallothionein inhibits peroxynitrite-induced DNA and lipoprotein damage. AB - Previous studies have demonstrated that metallothionein functions as an antioxidant that protects against oxidative DNA, protein, and lipid damage induced by superoxide anion, hydrogen peroxide, hydroxyl radical, and nitric oxide. The present study was undertaken to test the hypothesis that metallothionein also protects from DNA and lipoprotein damage induced by peroxynitrite, an important reactive nitrogen species that causes a diversity of pathological processes. A cell-free system was used. DNA damage was detected by the mobility of plasmid DNA in electrophoresis. Oxidation of low density lipoprotein was measured by a thiobarbituric acid-reactive substance, which was confirmed by lipid hydroperoxide assay. Plasmid DNA damage and low density lipoprotein oxidation were induced by 3-morpholinosydnomine, which produces peroxynitrite through the reaction between nitric oxide and superoxide anion or by synthesized peroxynitrite directly. DNA damage by 3-morpholinosydnomine was prevented by both metallothionein and superoxide dismutase, whereas the damage caused by peroxynitrite was prevented by metallothionein only. The oxidation of low density lipoprotein by 3-morpholinosydnomine and peroxynitrite was also significantly inhibited by metallothionein. This study thus demonstrates that metallothionein may react directly with peroxynitrite to prevent DNA and lipoprotein damage induced by this pathological reactive nitrogen species. PMID- 11042195 TI - Ca2+- and phosphatidylinositol 3-kinase-dependent nitric oxide generation in lung endothelial cells in situ with ischemia. AB - Endothelial cells generate nitric oxide (NO) in response to agonist stimulation or increased shear stress. In this study, we evaluated the effects of abrupt cessation of shear stress on pulmonary endothelial NO generation and its relationship to changes in intracellular Ca(2+). In situ endothelial generation of NO and changes in intracellular Ca(2+) in isolated, intact rat lungs were evaluated using fluorescence microscopy with diaminofluorescein diacetate, an NO probe, and Fluo-3, a Ca(2+) probe. The onset of increased NO generation in endothelial cells of subpleural microvessels in situ occurred between 30 and 90 s after onset of ischemia and was preceded by an increase in intracellular Ca(2+) due to both influx of extracellular Ca(2+) and release from intracellular stores. Flow cessation-induced NO generation in endothelial cells in situ was Ca(2+)-, calmodulin-, and PI3-kinase-dependent. The similarity of endothelial cell response (increased NO generation) to either increased flow or cessation of flow suggests that cells respond to an imposed alteration from a state of adaptation. This response to flow cessation may constitute a compensatory vasodilatatory mechanism and may play a role in signaling for cell proliferation and vascular remodeling. PMID- 11042197 TI - Characterization of homo- and heterodimerization of cardiac Csx/Nkx2.5 homeoprotein. AB - Csx/Nkx2.5 is an evolutionarily conserved homeodomain (HD)-containing transcription factor that is essential for early cardiac development. We found that the HD of Csx/Nkx2.5 binds as a monomer as well as a dimer to its DNA binding sites in the promoter of the atrial natriuretic factor (ANF) gene, an in vivo target gene of Csx/Nkx2.5. Csx/Nkx2.5 physically interacts with each other in vitro as well as in cells, and the HD is critical for homodimerization. Lys(193) and Arg(194), located at the COOH-terminal end of HD, are essential for dimerization. Lys(193) is also required for a specific interaction with the zinc finger transcription factor GATA4. Csx/Nkx2.5 can heterodimerize with other NK2 homeodomain proteins, Nkx2.3 and Nkx2.6/Tix, with different affinities. A single missense mutation, Ile(183) to Pro in the HD of Csx/Nkx2.5, preserved homodimerization function, but totally abolished DNA binding. Ile(183) --> Pro mutant acts in an inhibitory manner on wild type Csx/Nkx2.5 transcriptional activity through the ANF promoter in 10T1/2 cells. However, Ile(183) --> Pro mutant does not inhibit wild type Csx/Nkx2.5 function on the ANF promoter in cultured neonatal cardiac myocytes, possibly due to failure of dimerization in the presence of the target DNA. These results suggest that complex protein protein interactions of Csx/Nkx2.5 play a role in its transcriptional regulatory function. PMID- 11042196 TI - Induction of cytosolic phospholipase A2 by oncogenic Ras is mediated through the JNK and ERK pathways in rat epithelial cells. AB - Mutations in ras genes have been detected with high frequency in nonsmall cell lung cancer cells (NSCLC) and contribute to transformed growth of these cells. It has previously been shown that expression of oncogenic forms of Ras in these cells is associated with elevated expression of cytosolic phospholipase A(2) (cPLA(2)) and cyclooxygenase-2 (COX-2), resulting in high constitutive levels of prostaglandin production. To determine whether expression of constitutively active Ras is sufficient to induce expression of these enzymes in nontransformed cells, normal lung epithelial cells were transfected with H-Ras. Stable expression of H-Ras increased expression of cPLA(2) and COX-2 protein. Transient transfection with H-Ras increased promoter activity for both enzymes. H-Ras expression also activated all three families of MAP kinase: ERKs, JNKs, and p38 MAP kinase. Expression of constitutively active Raf did not increase either cPLA(2) or COX-2 promoter activity, but inhibition of the ERK pathway with pharmacological agents or expression of dominant negative ERK partially blocked the H-Ras-mediated induction of cPLA(2) promoter activity. Expression of dominant negative JNK kinases decreased cPLA(2) promoter activity in NSCLC cell lines and inhibited H-Ras-mediated induction in normal epithelial cells, whereas expression of constructs encoding constitutively active JNKs increased promoter activity. Inhibition of p38 MAP kinase or NF-kappaB had no effect on cPLA(2) expression. Truncational analysis revealed that the region of the cPLA(2) promoter from -58 to +12 contained sufficient elements to mediate H-Ras induction. We conclude that expression of oncogenic forms of Ras directly increases cPLA(2) expression in normal epithelial cells through activation of the JNK and ERK pathways. PMID- 11042198 TI - Subunit interactions of yeast NAD+-specific isocitrate dehydrogenase. AB - Yeast mitochondrial NAD(+)-specific isocitrate dehydrogenase is an octamer composed of four each of two nonidentical but related subunits designated IDH1 and IDH2. IDH2 was previously shown to contain the catalytic site, whereas IDH1 contributes regulatory properties including cooperativity with respect to isocitrate and allosteric activation by AMP. In this study, interactions between IDH1 and IDH2 were detected using the yeast two-hybrid system, but interactions between identical subunit polypeptides were not detected with this or other methods. A model for heterodimeric interactions between the subunits is therefore proposed for this enzyme. A corollary of this model, based on the three dimensional structure of the homologous enzyme from Escherichia coli, is that some interactions between subunits occur at isocitrate binding sites. Based on this model, two residues (Lys-183 and Asp-217) in the regulatory IDH1 subunit were predicted to be important in the catalytic site of IDH2. We found that individually replacing these residues with alanine results in mutant enzymes that exhibit a drastic reduction in catalysis both in vitro and in vivo. Also based on this model, the two analogous residues (Lys-189 and Asp-222) of the catalytic IDH2 subunit were predicted to contribute to the regulatory site of IDH1. A K189A substitution in IDH2 was found to produce a decrease in activation of the enzyme by AMP and a loss of cooperativity with respect to isocitrate. A D222A substitution in IDH2 produces similar regulatory defects and a substantial reduction in V(max) in the absence of AMP. Collectively, these results suggest that the basic structural/functional unit of yeast isocitrate dehydrogenase is a heterodimer of IDH1 and IDH2 subunits and that each subunit contributes to the isocitrate binding site of the other. PMID- 11042199 TI - Apolipoprotein E4 stimulates cAMP response element-binding protein transcriptional activity through the extracellular signal-regulated kinase pathway. AB - Inheritance of the epsilon4 allele of the apolipoprotein E gene (APOE4) is a major risk factor for the development of Alzheimer's disease (AD). Although the association between APOE4 and AD is well documented, the mechanism by which apolipoprotein E exerts an isoform-specific effect on neurons in disease is unknown. In this report, we demonstrate that apoE4 stimulates the transcriptional activity of cAMP-response element-binding protein (CREB) by activating the extracellular signal-regulated kinase (ERK) cascade in rat primary hippocampal neurons. In contrast, apoE3 was unable to stimulate CREB transcriptional activity and unable to activate the ERK pathway. Elevation of intracellular Ca(2+) levels are also involved because treatment with receptor-associated protein, nifedipine, MK801, removal of Ca(2+) from the medium and dantrolene all served to inhibit calcium elevation and attenuate the activation of CREB. Treatment with an apoE peptide was also found to facilitate transcription of the CREB-dependent genes, c fos and Bcl-2. In contrast to treatment with apoE3, our findings suggest apoE4 and apoE-peptide induce a novel signaling pathway. PMID- 11042200 TI - A role for the peroxin Pex8p in Pex20p-dependent thiolase import into peroxisomes of the yeast Yarrowia lipolytica. AB - Peroxins are proteins required for peroxisome assembly. The cytosolic peroxin Pex20p binds directly to the beta-oxidation enzyme thiolase and is necessary for its dimerization and peroxisomal targeting. The intraperoxisomal peroxin Pex8p has a role in the import of peroxisomal matrix proteins, including thiolase. We report the results of yeast two-hybrid analyses with various peroxins of the yeast Yarrowia lipolytica and characterize more fully the interaction between Pex8p and Pex20p. Coimmunoprecipitation showed that Pex8p and Pex20p form a complex, while in vitro binding studies demonstrated that the interaction between Pex8p and Pex20p is specific, direct, and autonomous. Pex8p fractionates with peroxisomes in cells of a PEX20 disruption strain, indicating that Pex20p is not necessary for the targeting of Pex8p to peroxisomes. In cells of a PEX8 disruption strain, thiolase is mostly cytosolic, while Pex20p and a small amount of thiolase associate with peroxisomes, suggesting the involvement of Pex8p in the import of thiolase after docking of the Pex20p-thiolase complex to the membrane. In the absence of Pex8p, peroxisomal thiolase and Pex20p are protected from the action of externally added protease. This finding, together with the fact that Pex8p is intraperoxisomal, suggests that Pex20p may accompany thiolase into peroxisomes during import. PMID- 11042201 TI - Functional characterization of vertebrate nonmuscle myosin IIB isoforms using Dictyostelium chimeric myosin II. AB - The alternatively spliced isoform of nonmuscle myosin II heavy chain B (MHC-IIB) with an insert of 21 amino acids in the actin-binding surface loop (loop 2), MHC IIB(B2), is expressed specifically in the central nervous system of vertebrates. To examine the role of the B2 insert in the motor activity of the myosin II molecule, we expressed chimeric myosin heavy chain molecules using the Dictyostelium myosin II heavy chain as the backbone. We replaced the Dictyostelium native loop 2 with either the noninserted form of loop 2 from human MHC-IIB or the B2-inserted form of loop 2 from human MHC-IIB(B2). The transformant Dictyostelium cells expressing only the B2-inserted chimeric myosin formed unusual fruiting bodies. We then assessed the function of chimeric proteins, using an in vitro motility assay and by measuring ATPase activities and binding to F-actin. We demonstrate that the insertion of the B2 sequence reduces the motor activity of Dictyostelium myosin II, with reduction of the maximal actin-activated ATPase activity and a decrease in the affinity for actin. In addition, we demonstrate that the native loop 2 sequence of Dictyostelium myosin II is required for the regulation of the actin-activated ATPase activity by phosphorylation of the regulatory light chain. PMID- 11042202 TI - Interactions of a novel inhibitor from an extremophilic Bacillus sp. with HIV-1 protease: implications for the mechanism of inactivation. AB - The active site cleft of the HIV-1 protease (PR) is bound by two identical conformationally mobile loops known as flaps, which are important for substrate binding and catalysis. The present article reports, for the first time, an HIV-1 PR inhibitor, ATBI, from an extremophilic Bacillus sp. The inhibitor is found to be a hydrophilic peptide with Mr of 1147, and an amino acid sequence of Ala-Gly Lys-Lys-Asp-Asp-Asp-Asp-Pro-Pro-Glu. Sequence homology exhibited no similarity with the reported peptidic inhibitors of HIV-1 PR. Investigation of the kinetics of the enzyme-inhibitor interactions revealed that ATBI is a noncompetitive and tight binding inhibitor with the IC(50) and K(i) values 18.0 and 17.8 nm, respectively. The binding of the inhibitor with the enzyme and the subsequent induction of the localized conformational changes in the flap region of the HIV-1 PR were monitored by exploiting the intrinsic fluorescence of the surface exposed Trp-42 residues, which are present at the proximity of the flaps. We have demonstrated by fluorescence and circular dichroism studies that ATBI binds in the active site of the HIV-1 PR and thereby leads to the inactivation of the enzyme. Based on our results, we propose that the inactivation is due to the reorganization of the flaps impairing its flexibility leading toward inaccessibility of the substrate to the active site of the enzyme. PMID- 11042203 TI - The N-terminal region of neuregulin isoforms determines the accumulation of cell surface and released neuregulin ectodomain. AB - Two neuregulin-1 isoforms highly expressed in the nervous system are the type III neuregulin III-beta1a and the type I neuregulin I-beta1a. The sequence of these two isoforms differs only in the region that is N-terminal of the bioactive epidermal growth factor-like domain. While the biosynthetic processing of the I beta1a isoform has been well characterized, the processing of III-beta1a has not been reported. In this study, we compared III-beta1a and I-beta1a processing. Both III-beta1a and I-beta1a were synthesized as transmembrane proproteins that were proteolytically cleaved to produce an N-terminal fragment containing the bioactive epidermal growth factor-like domain. For I-beta1a, this product was released into the medium. However, for III-beta1a, this product was a transmembrane protein. In cultures of cells expressing III-beta1a, the amount of neuregulin at the cell surface was much greater, and the amount in the medium was much less than in cultures expressing I-beta1a. Phorbol ester treatment and truncation of the cytoplasmic tail had markedly different effects on III-beta1a and I-beta1a processing. These results demonstrate an important role for the N terminal region in determining neuregulin biosynthetic processing and show that a major product of III-beta1a processing is a tethered ligand that may act as a cell surface signaling molecule. PMID- 11042204 TI - p38 Kinase-dependent MAPKAPK-2 activation functions as 3-phosphoinositide dependent kinase-2 for Akt in human neutrophils. AB - Akt activation requires phosphorylation of Thr(308) and Ser(473) by 3 phosphoinositide-dependent kinase-1 and 2 (PDK1 and PDK2), respectively. While PDK1 has been cloned and sequenced, PDK2 has yet to be identified. The present study shows that phosphatidylinositol 3-kinase-dependent p38 kinase activation regulates Akt phosphorylation and activity in human neutrophils. Inhibition of p38 kinase activity with SB203580 inhibited Akt Ser(473) phosphorylation following neutrophil stimulation with formyl-methionyl-leucyl-phenylalanine, FcgammaR cross-linking, or phosphatidylinositol 3,4,5-trisphosphate. Concentration inhibition studies showed that Ser(473) phosphorylation was inhibited by 0.3 microm SB203580, while inhibition of Thr(308) phosphorylation required 10 microm SB203580. Transient transfection of HEK293 cells with adenoviruses containing constitutively active MKK3 or MKK6 resulted in activation of both p38 kinase and Akt. Immunoprecipitation and glutathione S-transferase (GST) pull-down studies showed that Akt was associated with p38 kinase, MK2, and Hsp27 in neutrophils, and Hsp27 dissociated from the complex upon activation. Active recombinant MK2 phosphorylated recombinant Akt and Akt in anti-Akt, anti MK2, anti-p38, and anti-Hsp27 immunoprecipitates, and this was inhibited by an MK2 inhibitory peptide. We conclude that Akt exists in a signaling complex containing p38 kinase, MK2, and Hsp27 and that p38-dependent MK2 activation functions as PDK2 in human neutrophils. PMID- 11042205 TI - 7-ketocholesterol is an endogenous modulator for the arylhydrocarbon receptor. AB - We have identified 7-ketocholesterol (7-KC) as an endogenous modulator that inhibits transactivation by the arylhydrocarbon receptor (AhR) through competitive binding against xenobiotic ligands. 7-KC binds AhR and displaces labeled dioxin (2,3,7,8-tetrachlorodibenzo(p)dioxin (TCDD)). IC(50) is 5 x 10(-7) m in vivo and 7 x 10(-6) m in vitro. These figures are consistent with its concentration in human blood plasma and tissues. Association with 7-KC prevents AhR binding to DNA. 7-KC blocks the TCDD-mediated transactivation of stably expressed reporter gene constructs in T47-D cells as well as the expression of the endogenous CYP 1A1 gene in HepG2 cells and in primary porcine aortic endothelial cells. Injection of 7-KC to rats blocks the induction of CYP 1A1 messenger RNA and protein in endothelial cells from myocardial blood vessels. The differential sensitivity of mammalian species to toxic effects of AhR ligands, especially dioxin (TCDD), correlates with the expression of 7-hydroxycholesterol dehydrogenase, which synthesizes 7-KC from 7-hydroxycholesterol. The documented involvement of AhR ligands in cardiovascular diseases through lipid peroxidation and endothelium dysfunction can now be examined in the context of displacement of this protective modulator. PMID- 11042206 TI - The nonsense-mediated decay pathway and mutually exclusive expression of alternatively spliced FGFR2IIIb and -IIIc mRNAs. AB - Exons IIIb and IIIc of the FGFR2 gene are alternatively spliced in a mutually exclusive manner in different cell types. A switch from expression of FGFR2IIIb to FGFR2IIIc accompanies the transition of nonmalignant rat prostate tumor epithelial cells (DTE) to cells comprising malignant AT3 tumors. Here we used transfection of minigenes with and without alterations in reading frame and with and without introns to examine how translation affects observed FGFR2 splice products. We observed that nonsense mutations in other than the last exon led to a dramatic reduction in mRNA that is abrogated by removal of downstream introns in both DTE and AT3 cells. The mRNA, devoid of both IIIb and IIIc exons (C1-C2), is a major splice product from minigenes lacking an intron downstream of the second common exon C2. From these observations, we suggest that repression of exon IIIc and activation of exon IIIb inclusion in DTE cells lead to the generation of both C1-IIIb-C2 and C1-C2 products. However, the C1-C2 product from the native gene is degraded due to a frameshift and a premature termination codon caused by splicing C1 and C2 together. Derepression of exon IIIc and repression of exon IIIb lead to the generation of both C1-IIIc-C2 and C1-C2 products in AT3 cells, but the C1-C2 product is degraded. The C1-IIIb-IIIc-C2 mRNA containing a premature termination codon in exon IIIc was present, but at apparently trace levels in both cell types. The nonsense-mediated mRNA decay pathway and cell type dependent rates of inclusion of exons IIIb and IIIc result in the mutually exclusive expression of FGFR2IIIb and IIIc. PMID- 11042207 TI - Identification and biochemical characterization of an Arabidopsis indole-3-acetic acid glucosyltransferase. AB - Biochemical characterization of recombinant gene products following a phylogenetic analysis of the UDP-glucosyltransferase (UGT) multigene family of Arabidopsis has identified one enzyme (UGT84B1) with high activity toward the plant hormone indole-3-acetic acid (IAA) and three related enzymes (UGT84B2, UGT75B1, and UGT75B2) with trace activities. The identity of the IAA conjugate has been confirmed to be 1-O-indole acetyl glucose ester. A sequence annotated as a UDP-glucose:IAA glucosyltransferase (IAA-UGT) in the Arabidopsis genome and expressed sequence tag data bases given its similarity to the maize iaglu gene sequence showed no activity toward IAA. This study describes the first biochemical analysis of a recombinant IAA-UGT and provides the foundation for future genetic approaches to understand the role of 1-O-indole acetyl glucose ester in Arabidopsis. PMID- 11042208 TI - Odorants stimulate the ERK/mitogen-activated protein kinase pathway and activate cAMP-response element-mediated transcription in olfactory sensory neurons. AB - Olfactory sensory neurons (OSNs) respond acutely to volatile molecules and exhibit adaptive responses including desensitization to odorant exposure. Although mechanisms for short term adaptation have been described, there is little evidence that odorants cause long lasting, transcription-dependent changes in OSNs. Here we report that odorants stimulate cAMP-response element (CRE) mediated transcription in OSNs through Ca2+ activation of the ERK/MAPK/p90rsk pathway. Odorant stimulation of ERK phosphorylation was ablated by inhibition of calmodulin-dependent protein kinase II suggesting that odorant activation of ERK is mediated through this kinase. Moreover, a brief exposure in vivo to an odorant in vapor phase stimulated CRE-mediated gene transcription in discrete populations of OSNs. These data suggest that like central nervous system neurons, OSNs may undergo long term adaptive changes mediated through CRE-mediated transcription. PMID- 11042209 TI - The SH2 domain containing tyrosine phosphatase-1 down-regulates activation of Lyn and Lyn-induced tyrosine phosphorylation of the CD19 receptor in B cells. AB - SHP-1 is a cytosolic tyrosine phosphatase implicated in down-regulation of B cell antigen receptor signaling. SHP-1 effects on the antigen receptor reflect its capacity to dephosphorylate this receptor as well as several inhibitory comodulators. In view of our observation that antigen receptor-induced CD19 tyrosine phosphorylation is constitutively increased in B cells from SHP-l deficient motheaten mice, we investigated the possibility that CD19, a positive modulator of antigen receptor signaling, represents another substrate for SHP-1. However, analysis of CD19 coimmunoprecipitable tyrosine phosphatase activity in CD19 immunoprecipitates from SHP-1-deficient and wild-type B cells revealed that SHP-1 accounts for only a minor portion of CD19-associated tyrosine phosphatase activity. As CD19 tyrosine phosphorylation is modulated by the Lyn protein tyrosine kinase, Lyn activity was evaluated in wild-type and motheaten B cells. The results revealed both Lyn as well as CD19-associated Lyn kinase activity to be constitutively and inducibly increased in SHP-1-deficient compared with wild type B cells. The data also demonstrated SHP-1 to be associated with Lyn in stimulated but not in resting B cells and indicated this interaction to be mediated via Lyn binding to the SHP-1 N-terminal SH2 domain. These findings, together with cyanogen bromide cleavage data revealing that SHP-1 dephosphorylates the Lyn autophosphorylation site, identify Lyn deactivation/dephosphorylation as a likely mechanism whereby SHP-1 exerts its influence on CD19 tyrosine phosphorylation and, by extension, its inhibitory effect on B cell antigen receptor signaling. PMID- 11042210 TI - Two conserved lysines at the 50/20-kDa junction of myosin are necessary for triggering actin activation. AB - Actin stimulates myosin's activity by inducing structural alterations that correlate with the transition from a weakly to a strongly bound state, during which time inorganic phosphate (P(i)) is released from myosin's active site. The surface loop at the 50/20-kDa junction of myosin (loop 2) is part of the actin interface. Here we demonstrate that elimination of two highly conserved lysines at the C-terminal end of loop 2 specifically blocks the ability of heavy meromyosin to undergo a weak to strong binding transition with actin in the presence of ATP. Removal of these lysines has no effect on strong binding in the absence of nucleotide, on the rate of ADP binding or release, or on the basal ATPase activity. We further show that the 16 amino acids of loop 2 preceding the lysine-rich region are not essential for actin activation, although they do modulate myosin's affinity for actin in the presence of ATP. We conclude that interaction of the conserved lysines with acidic residues in subdomain 1 of actin either triggers a structural change or stabilizes a conformation that is necessary for actin-activated release of P(i) and completion of the ATPase cycle. PMID- 11042211 TI - Identification of glucosyltransferase genes involved in sinapate metabolism and lignin synthesis in Arabidopsis. AB - Sinapic acid is a major phenylpropanoid in Brassicaceae providing intermediates in two distinct metabolic pathways leading to sinapoyl esters and lignin synthesis. Glucosyltransferases play key roles in the formation of these intermediates, either through the production of the high energy compound 1-O sinapoylglucose leading to sinapoylmalate and sinapoylcholine or through the production of sinapyl alcohol-4-O-glucoside, potentially leading to the syringyl units found in lignins. While the importance of these glucosyltransferases has been recognized for more than 20 years, their corresponding genes have not been identified. Combining sequence information in the Arabidopsis genomic data base with biochemical data from screening the activity of recombinant proteins in vitro, we have now identified five gene sequences encoding enzymes that can glucosylate sinapic acid, sinapyl alcohol, and their related phenylpropanoids. The data provide a foundation for future understanding and manipulation of sinapate metabolism and lignin biology in Arabidopsis. PMID- 11042212 TI - Regulation of apoptosis by phosphatidylinositol 4,5-bisphosphate inhibition of caspases, and caspase inactivation of phosphatidylinositol phosphate 5-kinases. AB - Phosphoinositides such as phosphatidylinositol 3,4,5-trisphosphate and phosphatidylinositol 3,4-bisphosphate promote cell survival and protect against apoptosis by activating Akt/PKB, which phosphorylates components of the apoptotic machinery. We now report that another phosphoinositide, phosphatidylinositol 4,5 bisphosphate (PIP2) is a direct inhibitor of initiator caspases 8 and 9, and their common effector caspase 3. PIP2 inhibited procaspase 9 processing in cell extracts and in a reconstituted procaspase 9/Apaf1 apoptosome system. It inhibited purified caspase 3 and 8 activity, at physiologically attainable PIP2 levels in mixed lipid vesicles. Caspase 3 binding to PIP2 was confirmed by cosedimentation with mixed lipid vesicles. Overexpression of phosphatidylinositol phosphate 5-kinase alpha (PIP5KIalpha), which synthesizes PIP2, suppressed apoptosis, whereas a kinase-deficient mutant did not. Protection by the wild-type PIP5KIalpha was accompanied by decreases in the generation of activated caspases and of caspase 3-cleaved PARP. Protection was not mediated through PIP3 or Akt activation. An anti-apoptotic role for PIP(2) is further substantiated by our finding that PIP5KIalpha was cleaved by caspase 3 during apoptosis, and cleavage inactivated PIP5KIalpha in vitro. Mutation of the P(4) position (D279A) of the PIP5KIalpha caspase 3 cleavage consensus prevented cleavage in vitro, and during apoptosis in vivo. Significantly, the caspase 3-resistant PIP5KIalpha mutant was more effective in suppressing apoptosis than the wild-type kinase. These results show that PIP2 is a direct regulator of apical and effector caspases in the death receptor and mitochondrial pathways, and that PIP5KIalpha inactivation contributes to the progression of apoptosis. This novel feedforward amplification mechanism for maintaining the balance between life and death of a cell works through phosphoinositide regulation of caspases and caspase regulation of phosphoinositide synthesis. PMID- 11042213 TI - In vitro incorporation of nascent molybdenum cofactor into human sulfite oxidase. AB - We were able to reconstitute molybdopterin (MPT)-free sulfite oxidase in vitro with the molybdenum cofactor (Moco) synthesized de novo from precursor Z and molybdate. MPT-free human sulfite oxidase apoprotein was obtained by heterologous expression in an Escherichia coli mutant with a defect in the early steps of MPT biosynthesis. In vitro reconstitution of the purified apoprotein was achieved using an incubation mixture containing purified precursor Z, purified MPT synthase, and sodium molybdate. In vitro synthesized MPT generated from precursor Z by MPT synthase remains bound to the synthase. Surprisingly, MPT synthase was found capable of donating bound MPT to MPT-free sulfite oxidase. MPT was not released from MPT synthase when either bovine serum albumin or Moco-containing sulfite oxidase was used in place of aposulfite oxidase. After the inclusion of sodium molybdate in the reconstitution mixture, active sulfite oxidase was obtained, revealing that in vitro MPT synthase and aposulfite oxidase are sufficient for the insertion of MPT into sulfite oxidase and the conversion of MPT into Moco in the presence of high concentrations of molybdate. The conversion of MPT into Moco by molybdate chelation apparently occurs concomitantly with the insertion of MPT into sulfite oxidase. PMID- 11042214 TI - Biochemical analysis of the eIF2beta gamma complex reveals a structural function for eIF2alpha in catalyzed nucleotide exchange. AB - Eukaryotic translation initiation factor eIF2 is a heterotrimer that binds and delivers Met-tRNA(i)(Met) to the 40 S ribosomal subunit in a GTP-dependent manner. Initiation requires hydrolysis of eIF2-bound GTP, which releases an eIF2.GDP complex that is recycled to the GTP form by the nucleotide exchange factor eIF2B. The alpha-subunit of eIF2 plays a critical role in regulating nucleotide exchange via phosphorylation at serine 51, which converts eIF2 into a competitive inhibitor of the eIF2B-catalyzed exchange reaction. We purified a form of eIF2 (eIF2betagamma) completely devoid of the alpha-subunit to further study the role of eIF2alpha in eIF2 function. These studies utilized a yeast strain genetically altered to bypass a deletion of the normally essential eIF2alpha structural gene (SUI2). Removal of the alpha-subunit did not appear to significantly alter binding of guanine nucleotide or Met-tRNA(i)(Met) ligands by eIF2 in vitro. Qualitative assays to detect 43 S initiation complex formation and eIF5-dependent GTP hydrolysis revealed no differences between eIF2betagamma and the wild-type eIF2 heterotrimer. However, steady-state kinetic analysis of eIF2B catalyzed nucleotide exchange revealed that the absence of the alpha-subunit increased K(m) for eIF2betagamma.GDP by an order of magnitude, with a smaller increase in V(max). These data indicate that eIF2alpha is required for structural interactions between eIF2 and eIF2B that promote wild-type rates of nucleotide exchange. We suggest that this function contributes to the ability of the alpha subunit to control the rate of nucleotide exchange through reversible phosphorylation. PMID- 11042215 TI - Phylogenetic analysis of the UDP-glycosyltransferase multigene family of Arabidopsis thaliana. AB - A class of UDP-glycosyltransferases (UGTs) defined by the presence of a C terminal consensus sequence is found throughout the plant and animal kingdoms. Whereas mammalian enzymes use UDP-glucuronic acid, the plant enzymes typically use UDP-glucose in the transfer reactions. A diverse array of aglycones can be glucosylated by these UGTs. In plants, the aglycones include plant hormones, secondary metabolites involved in stress and defense responses, and xenobiotics such as herbicides. Glycosylation is known to regulate many properties of the aglycones such as their bioactivity, their solubility, and their transport properties within the cell and throughout the plant. As a means of providing a framework to start to understand the substrate specificities and structure function relationships of plant UGTs, we have now applied a molecular phylogenetic analysis to the multigene family of 99 UGT sequences in Arabidopsis. We have determined the overall organization and evolutionary relationships among individual members with a surprisingly high degree of confidence. Through constructing a composite phylogenetic tree that also includes all of the additional plant UGTs with known catalytic activities, we can start to predict both the evolutionary history and substrate specificities of new sequences as they are identified. The tree already suggests that while the activities of some subgroups of the UGT family are highly conserved among different plant species, others subgroups shift substrate specificity with relative ease. PMID- 11042216 TI - Negative cell cycle regulation and DNA damage-inducible phosphorylation of the BRCT protein 53BP1. AB - In a screen designed to discover suppressors of mitotic catastrophe, we identified the Xenopus ortholog of 53BP1 (X53BP1), a BRCT protein previously identified in humans through its ability to bind the p53 tumor suppressor. X53BP1 transcripts are highly expressed in ovaries, and the protein interacts with Xp53 throughout the cell cycle in embryonic extracts. However, no interaction between X53BP1 and Xp53 can be detected in somatic cells, suggesting that the association between the two proteins may be developmentally regulated. X53BP1 is modified via phosphorylation in a DNA damage-dependent manner that correlates with the dispersal of X53BP1 into multiple foci throughout the nucleus in somatic cells. Thus, X53BP1 can be classified as a novel participant in the DNA damage response pathway. We demonstrate that X53BP1 and its human ortholog can serve as good substrates in vitro as well as in vivo for the ATM kinase. Collectively, our results reveal that 53BP1 plays an important role in the checkpoint response to DNA damage, possibly in collaboration with ATM. PMID- 11042217 TI - Biogenesis of nonspecific lipid transfer protein and sterol carrier protein x: studies using peroxisome assembly-defective pex cell mutants. AB - Nonspecific lipid transfer protein (nsLTP; also called sterol carrier protein 2) with a molecular mass of 13 kDa is synthesized as a larger 15-kDa precursor (pre nsLTP) with an N-terminal 20-amino acid extension presequence, as well as with the peroxisome targeting signal type 1 (PTS1), Ala-Lys-Leu, at the C terminus. The precursor pre-nsLTP is processed to mature nsLTP by proteolytic removal of the presequence, most likely after being imported into peroxisomes. Sterol carrier protein x (SCPx), a 59-kDa branched-chain fatty acid thiolase of peroxisomes, contains the entire pre-nsLTP moiety at the C-terminal part and is converted to the 46-kDa form and nsLTP after the transport to peroxisomes. We investigated which of these two potential topogenic sequences functions in biogenesis of nsLTP and SCPx. Morphological and biochemical analyses, making use of Chinese hamster ovary cell pex mutants such as the PTS1 receptor-impaired pex5 and PTS2 import-defective pex7, as well as green fluorescent protein chimeras, revealed that both pre-nsLTP and SCPx are imported into peroxisomes by the Pex5p mediated PTS1 pathway. Nearly half of the pre-nsLTP remains in the cytosol, as assessed by subcellular fractionation of the wild-type Chinese hamster ovary cells. In an in vitro binding assay, only mature nsLTP, but not pre-nsLTP, from the cell lysates interacted with the Pex5p. It is likely, therefore, that modulation of the C-terminal PTS1 by the presequence gives rise to cytoplasmic localization of pre-nsLTP. PMID- 11042218 TI - Properties of the collagen type XVII ectodomain. Evidence for n- to c-terminal triple helix folding. AB - Collagen XVII is a transmembrane component of hemidesmosomal cells with important functions in epithelial-basement membrane interactions. Here we report on properties of the extracellular ectodomain of collagen XVII, which harbors multiple collagenous stretches. We have recombinantly produced subdomains of the collagen XVII ectodomain in a mammalian expression system. rColXVII-A spans the entire ectodomain from deltaNC16a to NC1, rColXVII-B is similar but lacks the NC1 domain, a small N-terminal polypeptide rColXVII-C encompasses domains deltaNC16a to C15, and a small C-terminal polypeptide rColXVII-D comprises domains NC6 to NC1. Amino acid analysis of rColXVII-A and -C demonstrated prolyl and lysyl hydroxylation with ratios for hydroxyproline/proline of 0.4 and for hydroxylysine/lysine of 0.5. A small proportion of the hydroxylysyl residues in rColXVII-C ( approximately 3.3%) was glycosylated. Limited pepsin and trypsin degradation assays and analyses of circular dichroism spectra clearly demonstrated a triple-helical conformation for rColXVII-A, -B, and -C, whereas the C-terminal rColXVII-D did not adopt a triple-helical fold. These results were further substantiated by electron microscope analyses, which revealed extended molecules for rColXVII-A and -C, whereas rColXVII-D appeared globular. Thermal denaturation experiments revealed melting temperatures of 41 degrees C (rColXVII A), 39 degrees C (rColXVII-B), and 35 degrees C (rColXVII-C). In summary, our data suggest that triple helix formation in the ectodomain of ColXVII occurs with an N- to C-terminal directionality. PMID- 11042219 TI - Phorbol ester-induced generation of reactive oxygen species is protein kinase cbeta -dependent and required for SAPK activation. AB - Treatment of human U-937 myeloid leukemia cells with 12-O-tetradecanoylphorbol-13 acetate (TPA) is associated with protein kinase C (PKC) betaII-mediated activation of the stress-activated protein kinase (SAPK) pathway. The present studies demonstrate that the TPA response of U-937 cells includes the generation of reactive oxygen species (ROS). By contrast, the TPA-resistant U-937 cell variant (TUR), which is deficient in PKCbetaII expression, failed to respond to TPA with the induction of ROS. Moreover, we show that TPA-induced ROS production is restored in TUR cells stably transfected to express PKCbetaII. The results also demonstrate that TPA-induced ROS production is required for activation of the MEK kinase-1 (MEKK-1)--> SAPK pathway. In concert with this observation, treatment of U-937 with H(2)O(2) as a source of ROS is associated with activation of the MEKK-1-->SAPK cascade. These findings indicate that PKCbetaII is required for TPA-induced ROS production and that the MEKK-1-->SAPK pathway is activated by a ROS-mediated mechanism. PMID- 11042220 TI - Up-regulation of nucleolin mRNA and protein in peripheral blood mononuclear cells by extracellular-regulated kinase. AB - The signal transduction pathways regulating nucleolin mRNA and protein production have yet to be elucidated. Peripheral blood mononuclear cells treated with phorbol 12-myristate 13-acetate showed steady state levels of nucleolin mRNA that were 2-2.5-fold greater than untreated control cells. The up-regulation of nucleolin mRNA was substantially repressed by U0126, a specific inhibitor that blocks phosphorylation of extracellular-regulated kinase (ERK). Calcium ionophores and ionomycin also activated ERK and substantially elevated nucleolin mRNA levels, demonstrating phorbol 12-myristate 13-acetate and calcium signaling converge on ERK. Drugs that affected protein kinase C, protein kinase A, and phospholipase C signal transduction pathways did not alter nucleolin mRNA levels significantly. The half-life of nucleolin mRNA increased from 1.8 h in resting cells to 3.2 h with phorbol ester activation, suggesting ERK-mediated posttranscriptional regulation. Concomitantly, full-length nucleolin protein was increased. The higher levels of nucleolin protein were accompanied by increased binding of a 70-kDa nucleolin fragment to the 29-base instability element in the 3'-untranslated region of amyloid precursor protein (APP) mRNA in gel mobility shift assays. Supplementation of rabbit reticulocyte lysate with nucleolin decreased APP mRNA stability and protein production. These data suggest ERK up regulates nucleolin posttranscriptionally thereby controlling APP production. PMID- 11042221 TI - Mutations that affect ligand binding to the Escherichia coli aspartate receptor: implications for transmembrane signaling. AB - Three arginine residues of the binding site of the Escherichia coli aspartate receptor contribute to its high affinity for aspartate (K(d) approximately 3 microm). Site-directed mutations at residue 64 had the greatest effect on aspartate binding. No residue could substitute for the native arginine; all changes resulted in an apparent K(d) of approximately 35 mm. These mutations had little impact on maltose responses. At residue Arg-69, a lysine substitution was least disruptive, conferring an apparent K(d) of 0.3 mm for aspartate. Results obtained for an alanine mutant were similar to those with cysteine and histidine mutants (K(d) approximately 5 mm) indicating that side chain size was not an important factor here. Proline and aspartate caused more severe defects, presumably for reasons related to conformation and charge. The impact of residue 69 mutations on the maltose response was small. Mutations at Arg-73 had similar effects on aspartate binding (K(d) 0.3-7 mm) but more severe consequences for maltose responses. Larger side chains resulted in the best aspartate binding, implying steric considerations are important here. Signaling in the mutant proteins was surprisingly robust. Given aspartate binding, signaling occurred with essentially wild-type efficiency. These results were evaluated in the context of available structural data. PMID- 11042222 TI - The zinc finger-containing transcription factors GATA-4, -5, and -6. Ubiquitously expressed regulators of tissue-specific gene expression. PMID- 11042223 TI - Induction and drug development. AB - Enzyme induction is an undesirable drug interaction, effecting the efficacy of co administered drugs, rather than safety. The number of clinically used drugs which induce P-450 enzymes is, in fact, quite limited. However, in certain disease areas (AIDS, epilepsy) many of the drugs used, whether for primary or secondary indications, have the potential for enzyme induction. Induction is often seen pre clinically, due to the elevated dose levels used, but this potential rarely transfers to the clinical situation. Some screening of induction potential can be conducted by the use of human hepatocytes, but supply and variable response limits their use. Robust and routine screens for enzyme induction are being developed based on nuclear receptors. However, whilst genuine structure activity relationships may emerge, with these new technologies, none is evident from the clinical information other than in general structures are diverse but most are lipophilic as defined by a positive calculated LogP value. A critical factor in P 450 induction in the clinic, based on the known drugs, is the question of dose size. The major inducible form of P-450 in man is CYP3A4. The drugs that induce CYP3A4 are given in high doses. In contrast to these amounts, many clinically used drugs, which are non-inducers are effective at doses up two orders of magnitude lower. As a first rule for drug discovery/development programmes it seems prudent to obey the "Golden Rules" of drug design: "Ensure moderate daily dose size by having chosen a viable mechanism and then increase potency against the target whilst optimising pharmacokinetics". This approach is exemplified by the antidiabetic compound troglitzone, a clinical CYP3A4 inducer which has a clinical dose of 200-600 mg and rosiglitazone a more potent analogue dose (2-12 mg) which is devoid of CYP3A4 induction in the clinic. PMID- 11042224 TI - Rational selection of antisense oligonucleotide sequences. AB - The purpose of this review is to identify rational selection procedures for the identification of optimal antisense oligonucleotide sequences. The review is firstly focused on how to find optimal hybridization sites, and secondly on how to select sequences that bind to structured RNA. The methods reviewed range from the more empirical testing of large numbers of mRNA complementary sequences to the more systematic techniques, i.e. RNase H mapping, use of combinatorial arrays and prediction of secondary structure of mRNA by computational methods. Structures that bind to structured RNA, i.e. aptastrucs and tethered oligonucleotide probes, and foldback triplex-forming oligonucleotides are also discussed. Relating to selection of antisense sequences by aid of computational analysis, valuable www addresses are given along with examples of folded structures of mRNA. PMID- 11042225 TI - Biodegradable pH-sensitive surfactants (BPS) in liposome-mediated nucleic acid cellular uptake and distribution. AB - The impact of biodegradable pH-sensitive surfactant (BPS)-liposomes on nucleic acid, i.e., oligonucleotide and plasmid DNA, cellular delivery was examined. Fluorescein-labeled nucleic acids complexed with 1,2-dioleoyl-3-trimethylammonium propane cationic liposomes and BPS at a charge ratio (+/-) of 10 were incubated in CV-1 cells and analyzed by flow cytometry. The fluorescence intensity of oligonucleotides but not plasmid DNA complexed with BPS-liposomes was higher than those complexed with BPS-free liposomes at early time points. However, when cells were fixed to equalize the intracellular pH since fluorescein, a pH-sensitive fluorophore, has higher fluorescence intensity in alkaline pH than acidic, no difference in intensity was observed. This indicated the incorporation of BPS in liposomes did not increase oligonucleotide cellular uptake over control liposomes, but redistributed oligonucleotides into a more basic environment, e.g., cytoplasm. An explanation consistent with the presented data is the formation of small transient membrane defects within the endosomal membrane as presented previously [Liang, E., Hughes, J.A., 1998a. Membrane fusion and rupture in liposomes: effect of biodegradable pH-sensitive surfactants. J. Membr. Biol. 166, 37-49.]. The above findings suggested that BPS may be effective agents of disrupting one of the major barriers, endosomal membrane, to enhance nucleic acid cellular transport. PMID- 11042227 TI - Pharmacokinetics of intravenously administered digoxin and histopathological picture in rabbits with experimental bile duct obstruction. AB - This study aimed to examine the effect of obstructive cholestasis on the pharmacokinetics of digoxin. Eighteen male rabbits were randomly ascribed to the two study groups: the sham-operated control group and the examined group - with common and cystic bile duct ligations. Digoxin was administered intravenously as a single dose of 0.02 mg/kg, and blood samples were withdrawn for up to 24 h. Digoxin concentrations were determined by the FPIA method. The pharmacokinetic parameters were calculated using a noncompartmental analysis. During the whole observation period the blood serum concentrations of digoxin were statistically higher in animals with obstructive cholestasis versus the controls. A significant increase in the area under the plasma concentration-time curve, decrease in the total body clearance and in the volume of distribution on the 6th day after the bile ducts ligation as compared to the sham-operated controls, were observed. The obtained results suggest an impaired elimination of digoxin in obstructive cholestasis in rabbits. PMID- 11042226 TI - Specificity of doxorubicin versus rhodamine-123 in assessing P-glycoprotein functionality in the LLC-PK1, LLC-PK1:MDR1 and Caco-2 cell lines. AB - The LLC-PK1:MDR1, LLC-PK1 and Caco-2 cell lines were used to investigate whether rhodamine-123 or doxorubicin would be the preferred substrate to study P glycoprotein (P-gp) functionality in vitro. Both rhodamine-123 and doxorubicin showed highly polarised transport in the Caco-2 cell line and the LLC-PK1:MDR1 cell line, indicating that P-gp is actively transporting these drugs. However, for rhodamine-123 polarised transport was also seen in the monolayers of the wild type LLC-PK1 cell line, indicating the presence of another active transporter for this compound. Polarised transport of doxorubicin in the Caco-2 and the LLC PK1:MDR1 cell lines could be inhibited by the P-gp inhibitors SDZ-PSC 833 (PSC 833), cyclosporin A (CsA), verapamil and quinine, but not by the inhibitors for the organic cation carrier systems cimetidine and tetraethylammonium (TEA). Polarised transport of rhodamine-123 in the Caco-2 cell line could only be inhibited by P-gp inhibitors. In the LLC-PK1:MDR1 and LLC-PK1 cell lines transport was also inhibited by inhibitors for the organic cation transport systems. In conclusion, rhodamine-123 is a substrate for both P-gp and the organic cation carrier systems in the kidney cell line. This indicates that rhodamine-123 is not selective enough to study P-gp functionality in cell systems were organic cation carrier systems are also present. Doxorubicin appears to be a more selective P-gp substrate and therefore more useful in studying P-gp functionality in vitro. PMID- 11042228 TI - Assessment of rate of drug release from oil vehicle using a rotating dialysis cell. AB - The rate constants for transfer of model compounds (naproxen and lidocaine) from oily vehicle (Viscoleo) to aqueous buffer phases were determined by use of the rotating dialysis cell. Release studies were done for the partly ionized compounds at several pH values. A correlation between the overall first-order rate constant related to attainment of equilibrium, k(obs), and the pH-dependent distribution coefficient, D, determined between oil vehicle and aqueous buffer was established according to the equation: logk(obs)=-0.71 logD-0.22 (k(obs) in h(-1)). Based on this correlation it was suggested that the rate constant of a weak electrolyte at a specified D value could be considered equal to the k(obs) value for a non-electrolyte possessing a partition coefficient, P(app), the magnitude of which was equal to D. Specific rate constants k(ow) and k(wo) were calculated from the overall rate constant and the pH-dependent distribution coefficient. The rate constant representing the transport from oily vehicle to aqueous phase, k(ow), was found to be significantly influenced by the magnitude of the partition coefficient P(app) according to: logk(ow)=-0.71 logP(app) log(P(app)+1)-0.22 (k(ow) in h(-1)). PMID- 11042229 TI - Modification of in vitro drug release rate from oily parenteral depots using a formulation approach. AB - Rate constants for transfer of naproxen and lidocaine from different oils and oil mixtures to aqueous buffer, pH 6.00, were determined using the rotating dialysis cell. Significantly different first-order rate constants related to attainment of equilibrium, k(obs), were derived depending on the type of oil/oil mixtures used in the release experiments. For the drugs a linear correlation was found between logk(obs) and the logarithm of the partition coefficient P(app): logk(obs)=-0.68 logP(app)-0.25 (k(obs) in h(-1), n=26). A linear relationship was observed between the calculated and experimentally determined P(app) values for the oil mixtures investigated. The specific rate constants, k(ow) and k(wo), related to the partition process were derived from the determined k(obs) and P(app) values. The rate constant k(ow) representing the rate of transfer of the solute from the oil phase to the aqueous buffer was shown to be strongly dependent on the partition coefficient according to the relationship: logk(ow)=-0.68 logP(app) log(P(app)+1)-0.25 (k(ow) in h(-1), n=26). In particular, diminished release rates were seen for oil mixtures containing castor oil most likely afforded by hydrogen bonding between the solute and the hydroxy groups of the latter vegetable oil. In this study it has been possible to alter P(app) for a specific compound up to a factor of 10 by variation of the composition of the oil vehicle. Such a span of P(app) values results in in vitro release rates differing a factor of 37. Thus, by proper design of the oil vehicle composition it should be possible to modify the release rate for a specific compound within certain limits. PMID- 11042230 TI - Supercritical fluid processing of proteins. I: lysozyme precipitation from organic solution. AB - The solution enhanced dispersion by supercritical fluid (SEDS) process was used to evaluate the effect of the processing variables on the biological and physicochemical characteristics of lysozyme protein particles produced from an organic solution of dimethylsulfoxide (DMSO) using an experimental design procedure. The processing variables were temperature, pressure, solution concentration and the flow-rates of supercritical carbon dioxide and a protein solution. Solutions of hen egg lysozyme (0.5-1%, w/v) in DMSO were dispersed using supercritical carbon dioxide as the antisolvent, and particles precipitated in a particle formation vessel. The morphology, particle size and size distribution and biological activity of the protein were determined. The precipitates were also examined with high sensitivity differential scanning calorimetry (HSDSC) and high-performance cation-exchange chromatography. The amount of residual DMSO was determined using headspace gas chromatography. Particle size measurements showed the precipitates to be agglomerates with primary particles of size 1-5 microm, containing <20 ppm of residual solvent. The activity of the precipitates varied between 44 and 100% depending on the experimental conditions. The similarity of HSDSC data for unprocessed and processed samples indicated that the SEDS process does not cause major denaturation of lysozyme when prepared from DMSO solutions. By optimising of working conditions, the SEDS process can produce micron-sized particles of lysozyme with minimal loss of biological activity. PMID- 11042231 TI - Population pharmacokinetics of tiagabine in epileptic patients on monotherapy. AB - A retrospective study of the population pharmacokinetics of tiagabine was performed from sparse data collected in a multicentre clinical trial in patients with newly diagnosed partial seizures. The purpose was to estimate the inter patient variability and to study the influence of various demographic, environmental and pathophysiological parameters on the pharmacokinetics of tiagabine in patients on monotherapy. A total of 593 plasma concentrations from 130 patients dosed with 2.5, 5, 7.5 or 10 mg tiagabine twice daily were used for modelling. A one-compartment open model with first-order absorption and elimination was fitted to the concentration-time data using the NONMEM program. Selection of covariates was initially performed using stepwise linear regression analyses. The selected covariates were incorporated in the population model and the importance of each covariate was investigated by means of backwards elimination. A one-compartment model with first-order absorption and elimination adequately described the tiagabine concentration-time profile. The apparent clearance as well as the apparent volume of distribution were both significantly correlated to body height in a nonlinear relationship. No other demographic, environmental or clinical chemical parameters were identified as covariates although only a few pathological values of the latter were present in the data. The mean values of CL/f was 6.10 l/h, of V/f was 62.0 l and of k(a) was 1.25 h( 1) for a subject of 170-cm height. The population half-life was 5.72 h. The apparent clearance and volume of distribution of tiagabine in epilepsy patients on monotherapy were both dependent on body height. Prospective studies are required in order to reveal if dose adjustments based on body height will result in improved therapeutic outcome. PMID- 11042232 TI - The results of a randomized trial of a quality improvement intervention in the care of patients with heart failure. The MISCHF Study Investigators. AB - PURPOSE: Quality improvement and disease management programs for heart failure have improved quality of care and patient outcomes at large tertiary care hospitals. The purpose of this study was to measure the effects of a regional, multihospital, collaborative quality improvement intervention on care and outcomes in heart failure in community hospitals. PATIENTS AND METHODS: This randomized controlled study included 10 acute care community hospitals in upstate New York. After a baseline period, 5 hospitals were randomly assigned to receive a multifaceted quality improvement intervention (n = 762 patients during the baseline period; n = 840 patients postintervention), while 5 were assigned to a "usual care" control (n = 640 patients during the baseline period; n = 664 patients postintervention). Quality of care was determined using explicit criteria by reviewing the charts of consecutive patients hospitalized with the primary diagnosis of heart failure during the baseline period and again in the postintervention period. Clinical outcomes included hospital length of stay and charges, in-hospital and 6-month mortality, hospital readmission, and quality of life measured after discharge. RESULTS: Patients had similar characteristics in the baseline and postintervention phases in the intervention and control groups. Using hospital-level analyses, the intervention had mixed effects on 5 quality-of care markers that were not statistically significant. The mean of the average length of stay among hospitals decreased from 8.0 to 6.2 days in the intervention group, with a smaller decline in mean length of stay in the control group (7.7 to 7.0 days). The net effects of the intervention were nonsignificant changes in length of stay of -1.1 days (95% confidence interval [CI]: -2.9 to 0.7 days, P = 0.18) and in hospital charges of -$817 (95% CI: -$2560 to $926, P = 0.31). There were small and nonsignificant effects on mortality, hospital readmission, and quality of life. CONCLUSIONS: The incremental effect of regional collaboration among peer community hospitals toward the goal of quality improvement was small and limited to a slightly, but not significantly, shorter length of stay. PMID- 11042233 TI - Effects of simulated altitude-induced hypoxia on exercise capacity in patients with chronic heart failure. AB - PURPOSE: Patients with stable heart failure often wish to spend time at altitudes above those of their residence. However, it is not known whether they can safely tolerate ascent to high altitudes or what its effects on work capacity may be. SUBJECTS AND METHODS: We studied 14 normal subjects and 38 patients with clinically stable heart failure, 12 of whom had normal workload [peak exercise oxygen consumption (VO(2)) greater than 20 mL/min/kg], 14 of whom had slightly diminished workload (peak VO(2) 20 to 15 mL/min/kg), and 12 of whom had markedly diminished workload (peak VO(2) less than 15 mL/min/kg) at baseline. All performed cardiopulmonary exercise tests with inspired oxygen fractions equal to those at 92, 1,000, 1,500, 2,000, and 3,000 m, and maximum achieved work rates (mean +/- SD) were measured. RESULTS: All subjects completed the trial; no test was interrupted because of arrhythmia, angina, or ischemia. Maximum work rate decreased in parallel with increasing simulated altitude. The percentage decrease was greater for patients with heart failure and was most marked among those with the lowest workload at baseline. Maximum achieved work rate declined by 3% +/- 4% per 1,000 m in normal subjects, by 5% +/- 3% (P <0.01) in patients with heart failure with normal workload, by 5% +/- 4% (P <0.01) in patients with slightly diminished workload, and by 11% +/- 5% (P <0.01 vs normal subjects and vs the other patients with heart failure) in patients with markedly reduced workload. CONCLUSION: Patients with stable heart failure who ascend to higher altitudes should expect to have a reduction in maximum physical activity in proportion to their exercise capacity at sea level. PMID- 11042234 TI - The association between the diameter of a patent foramen ovale and the risk of embolic cerebrovascular events. AB - PURPOSE: We sought to determine whether the size of a patent foramen ovale affected the risk of embolic cerebrovascular events of unknown origin. PATIENTS AND METHODS: We ascertained the presence and measured the size of patent foramen ovale using multiplane transesophageal echocardiography in 121 consecutive patients younger than 60 years who had transient ischemic attacks or ischemic strokes and in 123 control subjects. None of the patients had left heart, aortic, or carotid sources of embolism, or echocardiographic signs of elevated left or right atrial pressure. We used multivariate logistic regression to determine whether the size of the patent foramen ovale was an independent risk factor for cerebrovascular events. RESULTS: The mean (+/- SD) diameter of a patent foramen ovale was significantly larger in patients (4 +/- 2 mm) than in control subjects (2 +/- 1 mm, P <0.0001). A patent foramen ovale greater than 4 mm was associated with an increased risk of transient ischemic attacks [odds ratio (OR) = 3.4; 95% confidence interval (CI), 1.0 to 11, P = 0.04], ischemic strokes (OR = 12; 95% CI, 3.3 to 44, P = 0.0001), and, especially, having evidence of two or more strokes (OR = 27; 95% CI, 4.7 to 160, P = 0.0002). CONCLUSION: The diameter of a patent foramen ovale is an independent risk factor for ischemic events, especially recurrent strokes. PMID- 11042235 TI - The effects of transdermal estradiol on the response to mental stress in postmenopausal women: a randomized trial. AB - PURPOSE: Estrogens inhibit adrenomedullary catecholamine release and catecholamine-mediated responses to stress. We examined whether estrogen supplementation reduces the sympathoadrenal response to mental stress in postmenopausal women. MATERIALS AND METHODS: We compared the effects of 3-week treatment with transdermal 17-beta-estradiol and placebo in 10 postmenopausal women using a randomized, blinded, crossover design. We measured plasma catecholamine levels and the cardiovascular and metabolic responses to a 15 minute stress with mental arithmetic. Treatments were compared using repeated measures analysis of variance. RESULTS: During placebo treatment, mean (+/- SD) epinephrine levels reached a peak of 431 +/- 135 pmol/liter after 15 minutes of stress; the epinephrine response was blunted during estradiol treatment, with a peak of 357 +/- 77 pmol/liter (P <0.05). Estradiol also blunted the diastolic blood pressure response to stress (baseline levels of 78 +/- 15 mm Hg vs peak of 90 +/- 6 mm Hg during placebo; baseline of 80 +/- 8 mm Hg vs peak of 84 +/- 6 mm Hg during estradiol; P <0.05). Estradiol treatment also blunted the decrease in the standard deviation of the mean of the electrocardiographic RR intervals and the increase in the ratio between the low-frequency and high-frequency bandwidths. CONCLUSION: We observed a moderate, although significant, reduction in markers of the stress response to mental arithmetic in postmenopausal women treated with transdermal 17-beta-estradiol. PMID- 11042236 TI - An intensive communication intervention for the critically ill. AB - PURPOSE: We sought to determine the effects of a communication process that was designed to encourage the use of advanced supportive technology when it is of benefit, but to limit its burdens when it is ineffective. We compared usual care with a proactive, multidisciplinary method of communicating that prospectively identified for patients and families the criteria that would determine whether a care plan was effective at meeting the goals of the patient. This process allowed caregivers to be informed of patient preferences about continued advanced supportive technology when its continuation would result in a compromised functional outcome or death. MATERIALS AND METHODS: We performed a before-and after study in 530 adult medical patients who were consecutively admitted to a university tertiary care hospital for intensive care. Multidisciplinary meetings were held within 72 hours of critical care admission. Patients, families, and the critical care team discussed the care plan and the patients' goals and expectations for the outcome of critical care. Clinical "milestones" indicative of recovery were identified with time frames for their occurrence. Follow-up meetings were held to discuss palliative care options when continued advanced supportive technology was not achieving the patient's goals. We measured length of stay, mortality, and provider team and family consensus in 134 patients before the intensive communication intervention and in 396 patients after the intervention. RESULTS: Intensive communication significantly reduced the median length of stay from 4 days (interquartile range, 2 to 11 days) to 3 days (2 to 6 days, P = 0.01 by survival analysis). This reduction remained significant after adjustment for acute physiology and chronic health evaluation (APACHE) 3 score [risk ratio (RR) = 0.81; 95% confidence interval (CI), 0.66 to 0.99; P = 0.04). Subgroup analysis revealed that this reduction occurred in our target group, patients with acuity scores in the highest quartile who died (RR = 0.60; 95% CI, 0.38 to 0.92; P = 0.02). The intervention, which allowed dying patients earlier access to palliative care, was not associated with increased mortality. CONCLUSIONS: Intensive communication was associated with a reduction in critical care use by patients who died. Our multidisciplinary process targeted advanced supportive technology to patients who survived and allowed the earlier withdrawal of advanced supportive technology when it was ineffective. PMID- 11042237 TI - Are physicians aware of which of their patients have indwelling urinary catheters? AB - PURPOSE: Although infections associated with indwelling urinary catheters are common, costly, and morbid, the use of these catheters is unnecessary in more than one-third of patients. We sought to assess whether attending physicians, medical residents, and medical students are aware if their hospitalized patients have an indwelling urinary catheter, and whether physician awareness is associated with appropriate use of these catheters. METHODS: The physicians and medical students responsible for patients admitted to the medical services at four university-affiliated hospitals were given a list of the patients on their service. For each patient, the provider was asked: "As of yesterday afternoon, did this patient have an indwelling urethral catheter?" Respondents' answers were compared with the results of examining the patient. RESULTS: Among 288 physicians and students on 56 medical teams, 256 (89%) completed the survey. Of 469 patients, 117 (25%) had an indwelling catheter. There were a total of 319 provider-patient observations among these 117 patients. Overall, providers were unaware of catheterization for 88 (28%) of the 319 provider-patient observations. Unawareness rates by level of training were 21% for students, 22% for interns, 27% for residents, and 38% for attending physicians (P = 0.06). Catheter use was inappropriate in 36 (31%) of the 117 patients with a catheter. Providers were unaware of catheter use for 44 (41%) of the 108 provider-patient observations of patients who were inappropriately catheterized. Catheterization was more likely to be appropriate if respondents were aware of the catheter (odds ratio = 3.7; 95% confidence interval, 2.1 to 6.7, P <0.001). CONCLUSION: Physicians are commonly unaware that their patients have an indwelling urinary catheter. Inappropriate catheters are more often "forgotten" than appropriate ones. System wide interventions aimed at discontinuing unnecessary catheterization seem warranted. PMID- 11042238 TI - Management and dosing of warfarin therapy. AB - When initiating warfarin therapy, clinicians should avoid loading doses that can raise the International Normalized Ratio (INR) excessively; instead, warfarin should be initiated with a 5-mg dose (or 2 to 4 mg in the very elderly). With a 5 mg initial dose, the INR will not rise appreciably in the first 24 hours, except in rare patients who will ultimately require a very small daily dose (0.5 to 2.0 mg). Adjusting a steady-state warfarin dose depends on the measured INR values and clinical factors: the dose does not need to be adjusted for a single INR that is slightly out of range, and most changes should alter the total weekly dose by 5% to 20%. The INR should be monitored frequently (eg, 2 to 4 times per week) immediately after initiation of warfarin; subsequently, the interval between INR tests can be lengthened gradually (up to a maximum of 4 to 6 weeks) in patients with stable INR values. Patients who have an elevated INR will need more frequent testing and may also require vitamin K1. For example, a nonbleeding patient with an INR of 9 can be given low-dose vitamin K1 (eg, 2.5 mg phytonadione, by mouth). Patients who have an excessive INR with clinically important bleeding require clotting factors (eg, fresh-frozen plasma) as well as vitamin K1. PMID- 11042239 TI - Sex-related differences in the use and adverse effects of angiotensin-converting enzyme inhibitors in heart failure: the study of patients intolerant of converting enzyme inhibitors registry. PMID- 11042240 TI - The stress of being a doctor: steroid excretion rates in internal medicine residents on and off duty. PMID- 11042241 TI - Prescribing potassium despite hyperkalemia: medication errors uncovered by linking laboratory and pharmacy information systems. PMID- 11042242 TI - Central diabetes insipidus due to lymphocytic infundibuloneurohypophysitis. PMID- 11042243 TI - Diagnostic dilemma. PMID- 11042244 TI - Quality improvement studies: the need is there but so are the challenges. PMID- 11042245 TI - Taking heart failure to new heights: its pathophysiology at simulated altitude. PMID- 11042246 TI - Patent foramen ovale diameter and embolic stroke: a part of the puzzle? PMID- 11042247 TI - Answer to diagnostic dilemma PMID- 11042248 TI - Why Medicare supports graduate medical education. PMID- 11042249 TI - Dermal exposure: a decade of real progress. PMID- 11042250 TI - Dermal exposure assessment. AB - Assessing dermal exposure is a complex task. Even the most commonly used methods face fundamental problems and there are large gaps in the documentation and validation of sampling methods. Still larger uncertainties exist regarding strategies for measurement. We propose a strategy based on a conceptual model and which draws on the considerable insight gained for airborne contaminants, including EN 689 for assessing exposure by inhalation. The vast amount of air sampling data has provided good insight into the statistical properties of short term and long-term exposure levels, which is essential for designing cost effective exposure studies. For surface and skin contaminants an understanding of the distribution types and parameter values is only beginning to emerge. Transport rates away from the skin contaminant layer determine the 'memory' of a dermal sample and measurement principles are proposed depending on these rates. It is argued that uptake is the ultimate dermal exposure metric for risk assessment and should be the basis for devising dermal occupational exposure limits. PMID- 11042251 TI - Hand wash and manual skin wipes. AB - Hand wash and skin wipes are major techniques that have been used for dermal exposure sampling. Both techniques remove chemicals either deposited on or transferred to the skin contaminant layer by a combination of chemical and mechanical actions. The paper overviews identified methods and techniques, with emphasis on sampling parameters and sampling efficiency. It is concluded that identified sampling protocols, including sampling techniques, deviate at possible key issues, which hampers comparisons of study results. It is recommended to conduct sampling efficiency studies prior to field sampling, under conditions that are quite similar to conditions of exposure regarding exposure process, levels of skin loading, and time of residence of the compound on the skin. Harmonization of sampling protocols will be a first step in creating a database for better understanding the influence of sampling parameters on the performance of removal techniques to assess dermal exposure. PMID- 11042252 TI - Use of patches and whole body sampling for the assessment of dermal exposure. AB - There has been a growing awareness of the importance of dermal exposure in recent years. A wide range of techniques are employed to measure exposure, of which surrogate skin techniques such as patch sampling and whole body sampling are frequently used. One of the problems associated with dermal sampling is that different methods often produce different results due to differences in the principles involved in sample collection. As a consequence little progress towards establishing dermal exposure limits has been made. Both patches and clothing act as passive samplers and are intended to collect all of a substance deposited on them. This paper details the principles underlying patch and whole body sampling and outlines some of the advantages and disadvantages of each. A conceptual model has recently been proposed for dermal exposure and the role that surrogate techniques may play in the application of this model is discussed. Finally, suggestions are made as to how these techniques may be made more relevant and areas of future research highlighted. PMID- 11042253 TI - Use of qualitative and quantitative fluorescence techniques to assess dermal exposure. AB - Fluorescent tracers provide a way of simultaneously assessing the mass of a contaminant hazardous substance on the surface of the skin of a worker and the area of skin exposed. These parameters, along with the duration of exposure and the estimated contaminant concentration in the skin contamination layer, can be used to calculate the likely uptake through the skin. Repeated assessment of the mass of tracer on a surface within a room or on the surface of the skin can also allow the net transfer of contaminant to that compartment to be estimated. Qualitative evaluation of transfer processes using fluorescent tracers can help identify important secondary sources of exposure. PMID- 11042254 TI - Suction methods for assessing contamination on surfaces. AB - Suction sampling techniques are widely used to assess particulate contamination levels on domestic and occupational surfaces such as floor coverings, but their use for dermal exposure assessment has, to date, been limited. This paper reviews the sampling techniques commonly employed and summarises the range of sampling efficiencies reported in the literature. As there are an extremely large number of key factors influencing the recovery efficiency of suction sampling devices, it is recommended that controlled experiments are carried out to evaluate the relative significance of these factors, thus allowing inter-comparison of the data generated in field studies. As the range of applications of suction sampling devices is extensive, the harmonisation of sampling protocols is not considered to be a feasible objective. PMID- 11042255 TI - Modelling of skin exposure from distributed sources. AB - A simple model of indoor air pollution concentrations was used together with experimental results on deposition velocities to skin to calculate the skin dose from an outdoor plume of contaminants. The primary pathway was considered to be direct deposition to the skin from a homogeneously distributed air source. The model has been used to show that skin deposition was a significant dose contributor for example when compared to inhalation dose. PMID- 11042256 TI - Identification of dermal exposure pathways in the rubber manufacturing industry. AB - Current existing dermal exposure assessment strategies are predominantly based on regulatory protocols. In order to develop effective and efficient strategies more data driven approaches are needed. In a recently developed conceptual model for dermal exposure, compartments, barriers and mass transport processes relevant for dermal exposure were described. We systematically applied this conceptual model to the rubber manufacturing industry to assess dermal exposure to cyclohexane soluble matter (CSM) and used quantitative data to design an exposure assessment strategy. Identification of the spatial distribution of the dermal contamination showed high CSM surface concentrations for the upper body. Moreover, because of the high correlation between dermal exposure at the wrist and calculated total body exposure (r=0.89, P<0.01) an exposure assessment strategy based on only one pad sampler was employed to estimate CSM surface concentrations in the skin contaminant layer. Qualitative and quantitative evaluation of the relevant compartments and related mass transport processes demonstrated the importance of deposition of airborne contaminants and direct transfer of contaminants from sources and surfaces to the skin contaminant layer. Interestingly, the importance of the different exposure pathways varied considerably between production functions. The use of a model driven exposure assessment strategy in the rubber manufacturing industry revealed relevant skin regions, compartments and mass transport processes and enabled development of an effective and efficient strategy for dermal exposure assessment and hazard control in this particular occupational setting. PMID- 11042257 TI - Assessment of dermal exposure during airless spray painting using a quantitative visualisation technique. AB - The range of dermal exposure to non-volatile compounds during spray painting was studied in a semi-experimental study involving three enterprises and 12 painters. A fluorescent tracer was added to the paint and deposition of the tracer on clothing and uncovered parts of the skin was assessed using video imaging and processing techniques. A container (volume 36 m(3)) was sprayed with a colourless laquer (varnish) containing 66.7 mg/l fluorescent whitening agent. All painters sprayed the outside of the container. Nine painters repeated the painting a second time and five also sprayed the inside of the container. The painters wore white Tyvek coveralls, but no gloves. Duration of spraying the outside ranged from 4 to 21 min with a mean of 10 min and the amount of paint sprayed ranged from 3.0 to 12.8 l (mean 6.6 l). The mass of tracer deposited on the coverall ranged from 2.2 to 471 microg (90th percentile 256 microg), whereas, mass deposited on skin (i.e. the hands, wrists, and face) ranged from 0.01 to 52 microg tracer (90th percentile 20 microg). The quantity of tracer on the coverall was three times higher after spraying the inside of the container compared to spraying the outside, whereas the quantity on the skin was similar in both cases. On average 10% of the surface area of the coverall and skin was exposed during spraying the outside. Exposures, expressed in units of mass per area exposed were slightly higher for skin compared to coverall. In this study, deposited mass of tracer was correlated with an alternative exposure metric, i.e. surface area exposed multiplied by the duration of exposure, which has been proposed as a surrogate for uptake. Using a quantitative fluorescent tracer technique, it could be demonstrated that body parts which showed the lowest mass of tracer had the highest exposure as mass per surface area. Compared to other techniques which only determine mass, the ability to identify and quantify the actual surface area exposed is a clear advantage of the quantitative fluorescent tracer technique. PMID- 11042258 TI - Postulating a dermal pathway for exposure to anti-neoplastic drugs among hospital workers. Applying a conceptual model to the results of three workplace surveys. AB - Dermal exposure to anti-neoplastic drugs has been suggested as a potentially important route of exposure of hospital workers. Three small-scale workplace surveys were carried out in several hospitals focusing on contamination by leakage from IV infusion systems; contamination by spilled urine of patients treated with anti-neoplastic drugs and particulate phase anti-neoplastic drugs in the air of outpatient and nursing clinics. A new visual scoring technique using a fluorescent tracer was developed. The method showed a very low limit of detection (0.02 microl of contamination) and a very high inter-observer agreement (ICC=0.99). Evaluation of IV systems and connectors showed distinct differences between the systems. It was estimated that 0.5-250 microg of a drug can become available for contamination during each infusion. Differences in average contamination between nurses were negligible in the experimental set-up. Widespread and frequent contamination due to spillage of contaminated urine was revealed and appeared not to be restricted to the patient's room. Airborne particulate concentrations went undetected for 80% of the measurements. However, in a few cases concentrations up to 2 ng/m(3) of cyclophosphamide were measured predominantly in a room of a patient treated with this anti-neoplastic drug. Based on these results and a recently proposed conceptual model for dermal exposure a most likely exposure scenario was postulated both for nurses involved in administering drugs and nurses caring for treated patients. Estimation of all relevant mass transport rates will be a challenge for the near future. PMID- 11042259 TI - SGD1 encodes an essential nuclear protein of Saccharomyces cerevisiae that affects expression of the GPD1 gene for glycerol 3-phosphate dehydrogenase. AB - We here report the identification of the previously uncharacterized SGD1 gene, encoding a 102.8-kDa protein containing a leucine zipper region and a bipartite nuclear localization signal. Deletion of SGD1 results in loss of cell viability, while an increased dosage of SGD1 partially suppresses the osmosensitivity of pbs2 delta and hog1 delta mutants that are defective in the osmosignaling high osmolarity glycerol (HOG) mitogen-activated protein kinase pathway. The rescued mutants display a partially re-established transcriptional control of the osmostress-induced expression of GPD1, a target gene of the HOG pathway encoding NAD(+)-dependent glycerol 3-phosphate dehydrogenase, and a partially recovered hyperosmolarity-induced production of glycerol. Consistent with Sgd1p affecting the transcriptional control of GPD1, a functional green fluorescent protein tagged Sgd1p is localized to the cell nucleus. PMID- 11042260 TI - Genomic structure and promoter analysis of the rat kir7.1 potassium channel gene (Kcnj13). AB - In the brain inwardly rectifying potassium channel Kir7.1 subunits are predominantly expressed in the choroid plexus and meninges. To investigate this tissue-specific expression pattern, we characterized the genomic organization and the 5' proximal promoter of the rat Kir7.1 gene (Kcnj13). Starting from the major transcriptional initiation site, three exons in Kcnj13 give rise to the dominant approximately 1.45 kb transcript in brain. Adjacent to the transcriptional start the minimal promoter which, uncommon for ion channels, contains a TATA- and CAAT box is controlled by AP-1 factors and accounts for high gene expression levels. Luciferase reporter gene responses driven by the first 2.1 kb of the 5' flanking region were similarly high in epithelial FRTL-5 and neuronal N2A cells, suggesting that neuron-specific repressor elements are located remote from the non-selective minimal promoter. PMID- 11042261 TI - Oligomerization of the plasma membrane calcium pump involves two regions with different thermal stability. AB - Ca(2+) pump dimerization was studied by using a combined approach of thermal denaturation and fluorescence resonance energy transfer. The measurement of calcium pump ability to dimerize after the unfolding of individual functional domains of the enzyme demonstrated the existence of two different regions involved in the self-association process. One of these regions is highly susceptible to thermal unfolding and was identified as the calmodulin (CaM) binding domain. The other region whose thermal stability is higher than those of the catalytic and CaM-binding domains could be related with the previously found C28W-binding regions. PMID- 11042262 TI - A reversible protein phosphorylation system is present at the surface of infective larvae of the parasitic nematode Trichinella spiralis. AB - Trichinella spiralis infective larvae have externally oriented enzymes catalysing reversible protein phosphorylation on their surface. Incubation of larvae with exogenous ATP resulted in phosphorylation of surface bound and released proteins. Exposure of the parasites to bile, a treatment which renders them infective for intestinal epithelia, resulted in increased release of protein and an altered profile of phosphorylation. Both serine/threonine and tyrosine phosphorylation and dephosphorylation reactions took place at the parasite surface. Examination of the structural characteristics of the larvae following exposure to bile showed that the non-bilayer surface coat was not shed but was structurally reorganised. PMID- 11042263 TI - Evidence for the dual coupling of the rat neurotensin receptor with pertussis toxin-sensitive and insensitive G-proteins. AB - We previously demonstrated the functional coupling of the rat neurotensin receptor NTS1 with G-proteins on transfected CHO cell homogenates by showing modulation of agonist affinity by guanylyl nucleotides and agonist-mediated stimulation of [(35)S]GTP gamma S binding. In the present study, we observed that G(i/o)-type G-protein inactivation by pertussis toxin (PTx) resulted in a dramatic reduction of the NT-induced [(35)S]GTP gamma S binding whereas the effect of guanylyl nucleotide was almost not affected. As expected, NT-mediated phosphoinositide hydrolysis and intracellular calcium mobilization were not altered after PTx treatment. This suggests the existence of multiple signaling cascades activated by NT. Accordingly, using PTx and the PLC inhibitor U-73122, we showed that both signaling pathways contribute to the NT-mediated production of arachidonic acid. These results support evidence for a dual coupling of the NTS1 with PTx-sensitive and insensitive G-proteins. PMID- 11042264 TI - Direct binding of hepatitis B virus X protein and retinoid X receptor contributes to phosphoenolpyruvate carboxykinase gene transactivation. AB - The X gene product of the human hepatitis B virus (HBx), a major factor responsible for hepatitis and hepatocellular carcinoma, modulates transactivation by a variety of transcription factors. Herein, expression of the phosphoenolpyruvate carboxykinase (PEPCK) gene was found to be regulated transcriptionally by HBx through two distinct promoter regions. The cAMP response element (CRE)-1 site within the proximal promoter region mediated the HBx-induced transactivation of the PEPCK gene through C/EBP alpha and ATF-2. A retinoid X receptor (RXR) response element within the distal promoter region also contributed to the HBx-induced transactivation. Consistent with these results, HBx directly interacted with RXR, and the interaction interfaces were localized to the transactivation domain of HBx and the ligand binding domain of RXR. PMID- 11042265 TI - Regulation of tumor necrosis factor cytotoxicity by calcineurin. AB - Cyclosporin (CsA) inhibits mitochondrial death signaling and opposes tumor necrosis factor (TNF)-induced apoptosis in vitro. However, CsA is also a potent inhibitor of calcineurin, a phosphatase that may participate in cell death. Therefore, we tested the hypothesis that calcineurin regulates TNF cytotoxicity in rat hepatoma cells (FTO2B). TNF-treated FTO2B cells appeared apoptotic by DNA fragmentation, nuclear condensation, annexin V binding, and caspase activation. We studied two calcineurin inhibitors, CsA and FK506, and found that each potently inhibited TNF cytotoxicity. Western blot demonstrated calcineurin in FTO2B homogenates. In a model of mitochondrial permeability transition (MPT), we found that CsA prevented MPT and cytochrome c release, while FK506 inhibited neither. In summary, we present evidence that calcineurin participates in an apoptotic death pathway activated by TNF. CsA may oppose programmed cell death by inhibiting calcineurin activity and/or inhibiting mitochondrial signaling. PMID- 11042266 TI - Calcium channel beta subunits differentially modulate recovery of the channel from inactivation. AB - We examined the effects of calcium channel beta subunits upon the recovery from inactivation of alpha(1) subunits expressed in Xenopus oocytes. Recovery of the current carried by the L-type alpha(1) subunit (cyCa(v)1) from the jellyfish Cyanea capillata was accelerated by coexpression of any beta subunit, but the degree of potentiation differed according to which beta isoform was coexpressed. The Cyanea beta subunit was most effective, followed by the mammalian b(3), b(4), and beta(2a) subtypes. Recovery of the human Ca(v)2.3 subunit was also modulated by beta subunits, but was slowed instead. beta(3) was the most potent subunit tested, followed by beta(4), then beta(2a), which had virtually no effect. These results demonstrate that different beta subunit isoforms can affect recovery of the channel to varying degrees, and provide an additional mechanism by which beta subunits can differentially regulate alpha(1) subunits. PMID- 11042267 TI - Antioxidative galloyl esters as enzyme inhibitors of p-hydroxybenzoate hydroxylase. AB - Gallic acid and its esters were evaluated as enzyme inhibitors of recombinant p hydroxybenzoate hydroxylase (PHBH), a NADPH-dependent flavin monooxygenase from Pseudomonas aeruginosa. n-Dodecyl gallate (DG) (IC(50)=16 microM) and (-) epigallocatechin-3-O-gallate (EGCG) (IC(50)=16 microM), a major component of green tea polyphenols, showed the most potent inhibition, while product-like gallic acid did not inhibit the enzyme significantly (IC(50)>250 microM). Inhibition kinetics revealed that both DG and EGCG inhibited PHBH in a non competitive manner (K(I)=18.1 and 14.0 microM, respectively). The enzyme inhibition was caused by specific binding of the antioxidative gallate to the enzyme, and by scavenging reactive oxygen species required for the monooxygenase reaction. Molecular modeling predicted that EGCG binds to the enzyme in the proximity of the FAD binding site via formation of three hydrogen bonds. PMID- 11042268 TI - Induction of synthesis of an antimicrobial peptide in the skin of the freeze tolerant frog, Rana sylvatica, in response to environmental stimuli. AB - An extract of skin taken from specimens of the freeze-tolerant wood frog, Rana sylvatica, that were collected from cold (<7 degrees C) ponds and maintained at 5 degrees C lacked detectable antimicrobial activity. In contrast, an extract of skin taken from specimens maintained at 30 degrees C for 3 weeks under laboratory conditions contained a high concentration (approximately 4 nmol/g) of a single antimicrobial peptide of the brevinin-1 family (FLPVVAGLAAKVLPSIICAVTKKC). The peptide inhibited growth of Escherichia coli (minimum inhibitory concentration 45 microM) and Staphylococcus aureus (minimum inhibitory concentration 7 microM). The data suggest that synthesis of the peptide is induced when the animal is in an environment that promotes the growth of microorganisms consistent with a role in the animal's defense strategy. PMID- 11042269 TI - Identification of the histidine and aspartic acid residues essential for enzymatic activity of a family I.3 lipase by site-directed mutagenesis. AB - A lipase from Pseudomonas sp. MIS38 (PML) is a member of the lipase family I.3. We analyzed the roles of the five histidine residues (His(30), His(274), His(291), His(313), and His(365)) and five acidic amino acid residues (Glu(253), Asp(255), Asp(262), Asp(275), and Asp(290)), which are fully conserved in the amino acid sequences of family I.3 lipases, by site-directed mutagenesis. We showed that the mutation of His(313) or Asp(255) to Ala almost fully inactivated the enzyme, whereas the mutations of other residues to Ala did not seriously affect the enzymatic activity. Measurement of the far- and near-UV circular dichroism spectra suggests that inactivation by the mutation of His(313) or Asp(255) is not due to marked changes in the tertiary structure. We propose that His(313) and Asp(255), together with Ser(207), form a catalytic triad in PML. PMID- 11042270 TI - Photoreceptors and olfactory cells express the same retinal guanylyl cyclase isoform in medaka: visualization by promoter transgenics. AB - We examined the spatial expression patterns of two orphan receptor guanylyl cyclase genes OlGC4 and OlGC5 during embryogenesis of medaka and characterized the 5' flanking region required for tissue-specific expression of OlGC4 by introducing promoter-GFP fusion constructs into medaka embryos. Expression of OlGC5 is confined to retinal photoreceptor cells, while OlGC4 is expressed in the retina, pineal organ, and olfactory pits. The OlGC4 upstream region between -2374 and +343 is sufficient to drive the sensory organ-specific gene expression. Mutations in the consensus binding sequences for OTX/CRX transcription factors did not impair the reporter gene expression. Our results suggest that the same isoform of guanylyl cyclase is utilized in both photoreceptors and olfactory cells, and that transcription factors other than OTX/CRX primarily activate the OlGC4 expression. PMID- 11042271 TI - Pantetheinase activity of membrane-bound Vanin-1: lack of free cysteamine in tissues of Vanin-1 deficient mice. AB - Pantetheinase (EC 3.5.1.-) is an ubiquitous enzyme which in vitro has been shown to recycle pantothenic acid (vitamin B5) and to produce cysteamine, a potent anti oxidant. We show that the Vanin-1 gene encodes pantetheinase widely expressed in mouse tissues: (1) a pantetheinase activity is specifically expressed by Vanin-1 transfectants and is immunodepleted by specific antibodies; (2) Vanin-1 is a GPI anchored pantetheinase, and consequently an ectoenzyme; (3) Vanin-1 null mice are deficient in membrane-bound pantetheinase activity in kidney and liver; (4) in these organs, a major metabolic consequence is the absence of detectable free cysteamine; this demonstrates that membrane-bound pantetheinase is the main source of cysteamine in tissues under physiological conditions. Since the Vanin-1 molecule was previously shown to be involved in the control of thymus reconstitution following sublethal irradiation in vivo, this raises the possibility that Vanin/pantetheinase might be involved in the regulation of some immune functions maybe in the context of the response to oxidative stress. PMID- 11042272 TI - Hypochlorite modified LDL are a stronger agonist for platelets than copper oxidized LDL. AB - Experimental low density lipoprotein (LDL) oxidation is usually performed using trace copper, although the in vivo relevance of this method has been called into question. Such LDL augment adenosine 5'-diphosphate (ADP) induced platelet aggregation, presumably by the action of lipid derived compounds. In striking contrast, we find that LDL oxidized to a comparable extent by hypochlorite, an in vivo occurring oxidant, reveal themselves to be potent promoters of platelet aggregation. Interestingly, hypochlorite modified LDL seem to mediate their influence on human platelets by means of the modified apolipoprotein B-100 (apoB) moiety. Also, the finding that hypochlorite modified albumin is able to trigger platelet aggregation suggests an essential role for hypochlorite modified protein(s) in the process of platelet activation. PMID- 11042273 TI - A potential role for p15(Ink4b) and p57(Kip2) in liver development. AB - Hepatocytes undergo marked changes in proliferation during normal liver development. In order to elucidate the mechanism for these changes, we examined the ontogeny of expression for the known cyclin-dependent kinase inhibitors (CKIs), p15(Ink4b), p16(Ink4a), p18(Ink4c), p19(Ink4d), p21(Cip1), p27(Kip1) and p57(Kip2). All except p16(Ink4a) were expressed at some time between late gestation and adulthood. The mRNA and protein expression patterns for p15(Ink4b) and p57(Kip2) were consistent with a role for these CKIs in the regulation of hepatocyte proliferation. Specifically, p57(Kip2) may contribute to hepatocyte growth arrest that occurs in term fetuses, while p15(Ink4b) may contribute to the maintenance of adult hepatocytes in a quiescent state. These results assign a possible role to two CKIs not previously identified as involved in hepatocyte cell cycle control. PMID- 11042274 TI - Identification of triphosphoribosyl-dephospho-CoA as precursor of the citrate lyase prosthetic group. AB - The gamma-subunit of citrate lyase (EC 4.1.3.6) contains the prosthetic group 2' (5"-phosphoribosyl)-3'-dephospho-CoA and serves as an acyl carrier protein (ACP). We recently showed that in Escherichia coli the proteins CitG and CitX are essential for holo-ACP synthesis and provided evidence that CitG catalyzes the formation of a prosthetic group precursor from ATP and dephospho-CoA, which is subsequently attached via phosphodiester linkage to apo-ACP by CitX. Here we prove that CitG indeed catalyzes the conversion of ATP and dephospho-CoA to adenine and 2'-(5"-triphosphoribosyl)-3'-dephospho-CoA, the predicted precursor of the prosthetic group. Furthermore, this precursor was transferred by CitX to apo-ACP, yielding holo-ACP. Thus, our proposed mechanism for holo-ACP synthesis could be verified. PMID- 11042275 TI - Constitutive expression of a small heat-shock protein confers cellular thermotolerance and thermal protection to the photosynthetic apparatus in cyanobacteria. AB - The role of a small heat-shock protein (Hsp) in the acquisition of thermotolerance in cyanobacteria was investigated. Synechococcus sp. PCC 7942 was transformed with an expression vector carrying the coding sequence of the hspA gene encoding a small heat-shock protein from Synechococcus vulcanus under the control of the tac promoter. The transformant which was shown to constitutively express HspA displayed improved viability compared with the reference strain upon transfer from 30 to 50 degrees C in the light. When the heat shock was given in darkness, the survival rate in the reference strain increased greatly, approaching a level similar to that for the HspA expressing strain after heat shock in the light. Expression of HspA increased thermal resistance of photosystem II (PS II) and protected phycocyanin from heat-induced photobleaching. Our results are indicative of a central role for HspA in amelioration of the harmful effect of light during heat stress and identified the possible sites of action of the small Hsp in vivo to be the PS II complex and the light-harvesting phycobilisomes. PMID- 11042276 TI - Sequence and electrophysiological characterization of two insect-selective excitatory toxins from the venom of the Chinese scorpion Buthus martensi. AB - The two insecticidal peptides Bm32-VI and Bm33-I, isolated from the venom of the Chinese scorpion Buthus martensi induce paralytical symptoms typical of insect contractive toxins. They show, respectively, 74% and 77% homology with AaIT from Androctonus australis, comparable insecticidal activity and no vertebrate toxicity. Under voltage-clamp conditions, both toxins induced (1) an increased fast Na(+) current, (2) a shift in voltage dependence of Na(+) current activation, (3) the occurrence of a delayed current, and (4) a slow development of a holding current. Increased Na(+) conductance at negative potential values is responsible for axonal hyperexcitability and the contractive paralysis of insect prey. PMID- 11042277 TI - Kinetic analysis of novel multisubstrate analogue inhibitors of thymidine phosphorylase. AB - A kinetic analysis was performed for the novel 1-(8-phosphonooctyl)-6-amino-5 bromouracil and 1-(8-phosphonooctyl)-7-deazaxanthine inhibitors of Escherichia coli thymidine (dThd) phosphorylase (TPase). The structure of the compounds was rationally designed based on the available crystal structure coordinates of bacterial TPase. These inhibitors reversibly inhibited TPase. Kinetic analysis revealed that the compounds inhibited TPase in a purely competitive or mixed fashion not only when dThd, but also when inorganic phosphate (Pi), was used as the variable substrate. In contrast, the free bases 6-amino-5-bromouracil and 7 deazaxanthine behaved as non-competitive inhibitors of the enzyme in the presence of variable Pi concentrations while being competitive or mixed with respect to thymine as the natural substrate. Our kinetic data thus revealed that the novel 1 (8-phosphonooctyl)pyrimidine/purine derivatives are able to function as multisubstrate inhibitors of TPase, interfering at two different sites (dThd(Thy) and phosphate-binding site) of the enzyme. To our knowledge, the described compounds represent the first type of such multisubstrate analogue inhibitors of TPase; they should be considered as lead compounds for the development of mechanistically novel type of TPase inhibitors. PMID- 11042278 TI - Corrigendum to: sphingosylphosphorylcholine induces Ca(2+)-sensitization of vascular smooth muscle contraction: possible involvement of rho-kinase (FEBS 24156). PMID- 11042279 TI - Induction and detection of antibodies to squalene. AB - An enzyme-linked immunosorbent assay (ELISA) utilizing antigen coated on hydrophobic polyvinyldiene fluoride (PVDF) membranes is described for detecting antibodies that bind to squalene (SQE). Because of the prior lack of availability of validated antibodies to SQE, positive controls for the assay were made by immunization with formulations containing SQE to create monoclonal antibodies (mAbs) that reacted with SQE. Among eight immunogens tested, only two induced detectable murine antibodies to SQE: liposomes containing dimyristoyl phosphatidylcholine, dimyristoyl phosphatidylglycerol, 71% SQE, and lipid A [L(71% SQE+LA)], and, to a much lesser extent, an oil-in-water emulsion containing SQE, Tween 80, Span 85, and lipid A. In each case, lipid A served as an adjuvant, but neither SQE alone, SQE mixed with lipid A, liposomes containing 43% SQE and lipid A, nor several other emulsions containing both SQE and lipid A, induced antibodies that reacted with SQE. Monoclonal antibodies produced after immunizing mice with [L(71% SQE+LA)] served as positive controls for developing the ELISA. Monoclonal antibodies were produced that either recognized SQE alone but did not recognize squalane (SQA, the hydrogenated form of SQE), or that recognized both SQE and SQA. As found previously with other liposomal lipid antigens, liposomes containing lipid A also induced antibodies that reacted with the liposomal phospholipids. However, mAbs were also identified that reacted with SQE on PVDF membranes, but did not recognize either SQA or liposomal phospholipid. The polyclonal antiserum produced by immunizing mice with [L(71% SQE+LA)] therefore contained a mixed population of antibody specificities and, as expected, the ELISA of polyclonal antiserum with PVDF membranes detected antibodies both to SQE and SQA. We conclude that SQE is a weak antigen, but that antibodies that specifically bind to SQE can be readily induced by immunization with [L(71% SQE+LA)] and detected by ELISA with PVDF membranes coated with SQE. PMID- 11042280 TI - A method for the production of cryopreserved aliquots of antigen-preloaded, mature dendritic cells ready for clinical use. AB - Dendritic cells (DC) are increasingly used as a vaccine. Unfortunately, a satisfactory cryopreservation of DC in the absence of FCS is not yet available, so that laborious repeated generation of DC from fresh blood or frozen peripheral blood mononuclear cells for each vaccination has been required to date. We now aimed at developing an effective cryopreservation method, and by testing several variables found that it was crucial to combine the most advantageous maturation stimulus with an improved freezing procedure. We generated monocyte-derived DC from leukapheresis products by using GM-CSF and IL-4 and showed that amongst several known maturation stimuli the cocktail consisting of TNF-alpha+IL-1 beta+IL-6+PGE(2) achieved the highest survival of mature DC. We then systematically explored cryopreservation conditions, and found that freezing matured DC at 1 degrees C/min in pure autologous serum+10% DMSO+5% glucose at a cell density of 10x10(6) DC/ml gave the best results. Using this approach 85-100% of the frozen DC could be recovered in a viable state after thawing (Table 1). The morphology, phenotype, survival as well as functional properties (allogeneic mixed leukocyte reaction, induction of influenza matrix or melan A peptide specific cytotoxic T cells) of these thawed DC were equivalent to freshly prepared ones. The addition of CD40L or TRANCE/RANKL further improved DC survival. Importantly, we demonstrate that DC can effectively be loaded with antigens (such as Tetanus Toxoid, influenza matrix and melan A peptides) before cryopreservation so that it is now possible to generate antigen-preloaded, frozen DC aliquots that after thawing can be used right away. This is an important advance as both the generation of a standardized DC vaccine under GMP conditions and the carrying out of clinical trials are greatly facilitated. PMID- 11042281 TI - Thymocyte differentiation from lentivirus-marked CD34(+) cells in infant and adult human thymus. AB - Changes in thymic function and immune system homeostasis associated with HIV infection or chemotherapy have significant effects on the ability of patients to maintain a complete T cell receptor repertoire. Therefore, the development of in vitro systems to evaluate thymic function in children and adults may aid in the understanding of thymopoiesis and the development of new therapies to improve thymic output. Here we use a lentivirus-based gene transfer system to mark CD34(+) cells with EGFP and follow their differentiation into CD4(+) and CD8(+) single positive thymocytes in human thymic organ cultures. Lentivirus-marked cells entered the thymus and were detected in both the cortex and medulla. Pretreatment of the thymus with 2-deoxyguanosine depleted resident thymocytes and significantly increased the percentage of EGFP(+) thymocytes. High frequency gene transfer into CD34(+) cells and maintained expression throughout differentiation allows for the in vitro assessment of thymic function. In thymuses ranging in age from fetal to adult we observed EGFP(+) thymocytes at all stages of development suggesting that thymuses of all ages are capable of accepting new T cell progenitors and contributing to the maintenance of T cell homeostasis. PMID- 11042283 TI - Comparison of four serological methods for the detection of diphtheria anti-toxin antibody. AB - The aim of this study was to compare four serological methods for the detection of Corynebacterium diphtheriae IgG anti-toxin antibodies (IgG-DTAb) in human serum. One hundred serum samples were evaluated for C. diphtheriae IgG-DTAb by four different methods: passive haemagglutination (PHA), latex agglutination test (LA), toxoid enzyme-linked immunosorbent assay (Toxoid-ELISA), and toxin-binding inhibition enzyme-linked immunosorbent assay (ToBI-ELISA). As the external standardisation the neutralisation test for C. diphtheriae toxin in Vero cells (TN Vero) was used. For internal standardisation of IgG-DTAb titres, the WHO standard serum of human diphtheria antitoxin was used. The study revealed a poor correlation between the reference test and the PHA (r=0.34 Pearson's correlation coefficient), an acceptable correlation for the LA (r=0.74), a good correlation for the Toxoid-ELISA (r=0.81) and a very good correlation for ToBI-ELISA (r=0.93). The sensitivity measurements of PHA, LA, Toxoid-ELISA and ToBI-ELISA tests, were 14, 100, 94, 96% respectively and the corresponding specificity characteristics were 86, 76, 94, 90 respectively. Of the four evaluated methods, the ToBI-ELISA could be recommended for scientific and precise laboratory assays of diphtheria antibody levels in humans. For screening purposes the Toxoid-ELISA could be used, but the accuracy of antibody titres below 0.1 IU/ml, considered as the limits of protection, is questionable. Both tests offer very useful alternatives to the in vitro diphtheria toxin neutralisation test in Vero cells. Because of their unsatisfactory correlation and sensitivity as compared to the reference method, PHA and LA should be avoided and replaced by one of the two enzyme immunoassays. PMID- 11042282 TI - A quantitative flow cytometry assay for the preclinical testing and pharmacological monitoring of rabbit antilymphocyte globulins (rATG). AB - Rabbit antilymphocyte or antithymocyte globulins (rATG) are currently used as immunosuppressive agents in clinical organ and bone marrow transplantation, but because of differences in antigen source and purification processes between manufacturers, determination of the dose to be administered remains empirical. Dosage may be adjusted to peripheral blood T cell counts or limited by side effects such as neutropenia or thrombocytopenia. We report here a measurement by indirect immunofluorescence of the amount of antibodies from rATG that bind to different blood cell types. A concentration-related immunofluorescence signal was obtained with reasonable replicability, allowing comparison of different rATGs by reference to the same rATG batch as internal standard. Ratios of antilymphocyte to undesirable antibodies (directed against antigens expressed on neutrophils, platelets or red cells) could be calculated for each rATG preparation. Finally, the method is applicable to the measurement of free antibodies in sera from patients treated with rATG and to the analysis of the effects of in vivo absorption of rATG during its administration. Marked differences in antibody content per mg of rIgG account for the differences in rATG dosage that were empirically established for clinical applications. PMID- 11042284 TI - Isolation and tissue profiles of a large panel of phage antibodies binding to the human adipocyte cell surface. AB - Phage display is a powerful technique for the rapid selection and isolation of antibodies to any given target antigen. We have applied this technology to isolate over 100 different human antibodies that bind to antigens expressed in situ on the human adipocyte cell surface. This is a diverse panel of antibodies, as indicated by the V-region sequences. The binding profile of each anti adipocyte antibody has been characterised using phage antibody immunocytochemistry against a panel of normal human tissues. Although there was some variation in the intensity of the adipocyte staining, each antibody consistently recognised adipocytes, where present, irrespective of the tissue source. In addition, all of the antibodies recognised at least one other cell type other than the adipocyte cell surface. In total, over 50 different tissue binding profiles were recorded, with the most frequently recognised tissues identified as capillaries or smooth muscle. Extensive tissue binding profiles were generated for some antibodies using a panel of 37 different human tissues. This identified anti-adipocyte antibodies with unexpected profiles, such as FAT.13, which binds only to adipocytes and capillaries in the entire tissue panel. We believe this is the most extensive survey ever undertaken of the human adipocyte cell surface. Moreover, similar methodology could be used to derive complete tissue-binding profiles of antibodies against cell-surface antigens of any cell type. Indeed, by screening antibodies on both normal and diseased tissues, it may be possible to identify antigenic associations between different cell types and the pathologies of many diseases. PMID- 11042286 TI - Freezing induces artificial cleavage of apoptosis-related proteins in human bone marrow cells. AB - The aim of this study was to investigate whether freeze-thawing of freshly isolated human mononuclear bone marrow cells (MNC) influences the integrity of apoptosis-related proteins as determined by immunoblot analyses. Our results show that bone marrow is more sensitive to this process than either myelomonocytoid leukemic P39 or Jurkat T-lymphocyte cell lines. Specifically, bone marrow cells displayed a high level of intrinsic proteolytic activity in response to a single freeze-thaw cycle, which led to the cleavage of various proteins involved in apoptosis cell signaling. This effect was completely blocked by the inclusion of broad-spectrum protease inhibitors in the freezing medium and subsequently thawing the cells on ice. Since differences in the freezing conditions (-80 degrees C vs. liquid nitrogen) did not alter the proteins of interest, we suggest that the thawing process is the critical point when proteolytic enzyme activity is elevated. PMID- 11042285 TI - Effect of IFN-gamma on the killing of S. aureus in human whole blood. Assessment of bacterial viability by CFU determination and by a new method using alamarBlue. AB - Given the increasing incidence of methicillin resistant Staphylococcus aureus (MRSA) and the recent emergence of MRSA with a reduced susceptibility to vancomycin, alternative approaches to the treatment of infection are of increasing relevance. The purpose of these studies was to evaluate the effect of IFN-gamma on the ability of white blood cells to kill S. aureus and to develop a simpler, higher throughput bacterial killing assay. Using a methicillin sensitive clinical isolate of S. aureus, a clinical isolate of MRSA, and a commercially available strain of MRSA, studies were conducted using a killing assay in which the bacteria were added directly into whole blood. The viability of the bacteria in samples harvested at various time points was then evaluated both by the classic CFU assay and by a new assay using alamarBlue. In the latter method, serially diluted samples and a standard curve containing known concentrations of bacteria were placed on 96-well plates, and alamarBlue was added. Fluorescence readings were taken, and the viability of the bacteria in the samples was calculated using the standard curve. The results of these studies demonstrated that the CFU and alamarBlue methods yielded equivalent detection of bacteria diluted in buffer. For samples incubated in whole blood, however, the alamarBlue method tended to yield lower viabilities than the CFU method due to the emergence of a slower growing subpopulation of S. aureus upon incubation in the blood matrix. A significant increase in bacterial killing was observed upon pretreatment of whole blood for 24 h with 5 or 25 ng/ml IFN-gamma. This increase in killing was detected equivalently by the CFU and alamarBlue methods. In summary, these studies describe a method that allows for the higher throughput analysis of the effects of immunomodulators on bacterial killing. PMID- 11042287 TI - Monoclonal antibodies generated against an affinity-labeled immune complex of an anti-bile acid metabolite antibody: an approach to noncompetitive hapten immunoassays based on anti-idiotype or anti-metatype antibodies. AB - Conventional immunoassays for haptens such as steroids and synthetic drugs are dependent on the competitive reaction between an unlabeled antigen (analyte) and a labeled antigen against a limited amount of anti-hapten antibody. Although noncompetitive immunoassay procedures such as two-site immunometric assays offer a much higher sensitivity, direct application of this principle to haptens has been difficult due to their small molecular mass precluding simultaneous binding by two antibody molecules. Here, we have attempted to develop a noncompetitive immunoassay system based on anti-idiotype or anti-metatype antibodies. Ursodeoxycholic acid 7-N-acetylglucosaminide (UDCA 7-NAG), which is a bile acid metabolite (molecular weight, 595.8), was selected as the model hapten. A/J mice were immunized with a monoclonal antibody against UDCA 7-NAG, which had been affinity-labeled with a relevant hapten derivative. The fusion between the immune spleen cells and P3/NS1/1-Ag4-1 myeloma cells yielded four kinds of alpha-type and two kinds of beta-type monoclonal anti-idiotype antibodies, each recognizing the framework region and paratope of the anti-hapten antibody. The use of a selected combination between alpha-type and beta-type antibodies together with the anti-hapten antibody provided a noncompetitive assay system with a subfemtomole order sensitivity (detection limit, 118 amol) and a practical specificity. PMID- 11042288 TI - Biolistic-mediated gene transfer using the bovine herpesvirus-1 glycoprotein D is an effective delivery system to induce neutralizing antibodies in its natural host. AB - A genetic vaccine consisting of the bovine herpesvirus-1 (BHV-1) glycoprotein D (gD) gene was constructed and administered to cattle using the biolistic (gene gun) process. Results were compared to standard intramuscular injection of an inactivated whole BHV-1 commercial vaccine. Cattle genetically immunized by the gene-gun-delivered gD subunit vaccine developed high titers of IgG antibodies specific to gD demonstrating that this immunization method is a potent humoral response inducer. Further, gene-gun vaccinated cattle produced high neutralizing antibody titers to BHV-1 similar to levels induced in the commercial vaccine immunized animals. Additionally, cellular immunity was measured by an increased level of IFN-gamma mRNA detected in PBMC of cattle immunized with the gD gene or with the commercial vaccine, whereas augmented levels of IL-4 were not detected following vaccination. Because of its simplicity and effectiveness in inducing an immune response in cattle similar to a commercial vaccine, gene-gun delivery of a subunit BHV-1 gD vaccine would be a viable alternative to current immunization protocols. PMID- 11042290 TI - A cassette vector for high-level reporter expression driven by a hybrid invariant chain promoter in transgenic mice. AB - A plasmid cassette vector was designed to generate transgenic mice expressing reporter cDNAs at high levels in antigen-presenting cells under the control of the murine invariant chain (Ii) promoter. Analysis of several transgenic mice harboring a chimeric Ii cDNA placed in this vector showed that it can drive expression of the reporter protein to levels comparable to those of endogenous Ii. Furthermore, its expression pattern overlaps quite well with that of endogenous Ii. This vector should therefore be a convenient and versatile tool for the generation of transgenic mouse lines in which a desired protein may be expressed in Ii-positive cells at levels similar to those of endogenous Ii. Such a vector would be ideal for complementation studies of Ii-deficiency by specific Ii variants. PMID- 11042289 TI - Recombinant antibodies as carrier proteins for sub-unit vaccines: influence of mode of fusion on protein production and T-cell activation. AB - A major objective in development of vaccines is the design of sub-unit vaccines with the ability to induce strong T-cell responses. For this purpose, T-cell epitopes have been genetically inserted into various carrier proteins. Ig molecules may be especially useful as vehicles for delivery of CD4(+) T-cell epitopes to antigen presenting cells (APC). We have previously replaced loop structures between beta-strands in the C(H)1 domain of human IgG3 with a defined 11 amino acids long, MHC class II-restricted T-cell epitope. In this report we have added the same T-cell epitope into loops in the C(H)1 domain of mouse IgG2b. The following major points can be made: (1) Loops can accommodate an elongation of at least 11 amino acids without disruption of the overall Ig structure and secretion. (2) The recombinant Ig molecules are processed by spleen APC and the epitopes that are released are presented to T-cells. (3) Site of integration influences efficiency of processing and presentation. (4) Elongation of two neighbouring loops reduces Ig secretion. Taken together, our present results indicate that IgG C(H)1 domains may be engineered to carry T-cell epitopes in loop structures between beta-strands, but not all loops may be equally suitable for this purpose. PMID- 11042292 TI - A simple and rapid flow cytometric method for detection of porcine cell surface markers. AB - The objective of this study was to develop a rapid and reliable method for flow cytometric analysis of porcine whole blood cells. Fifty-microliters of heparin- or EDTA-treated whole blood was added to wells of a round-bottom 96-well microtitration plate. Each well contained 10 microl of an appropriate dilution of four different antibodies (40 microl total; two primary monoclonal antibodies and two fluorescent-labeled secondary antibodies). For convenience, the antibody mixture could be added to plates 1-2 days prior to assay and stored at 4 degrees C. Once whole blood was added to wells, plates were mixed gently, placed in a sealed bag and incubated in the dark at room temperature for 20 min. Contents of wells were then transferred to polystyrene tubes containing 2 ml of 1.5% formalin in distilled water and mixed gently. Cells were fixed for a minimum of 30 min and then stored in the dark at 4 degrees C until analysis by flow cytometry. Analysis of cell samples may be done up to 3 days after fixation. Results indicate that the percentages of Class I, Class II, CD3, CD8, CD4, CD45, monocyte, gamma-delta T-cell populations, and total number of granulocytes identified using this method were comparable to standard values or to values obtained following separation of white blood cells from red blood cells. The percentage of labeled B-cells was lower than standard values. Total assay time from receipt of blood to acquisition of data by flow cytometry required less than 2 h. This modified assay was shown to be simple, reliable, and useful for screening large numbers of porcine samples in a minimal period of time. PMID- 11042291 TI - Analysis of receptor/ligand interactions using whole-molecule randomly-mutated ligand libraries. AB - We report a novel method for the analysis of protein ligands using a whole molecule mutagenesis/phage display system. The cDNA for the inflammatory polypeptide C5a was used as template in a PCR reaction doped with mutagenic nucleoside triphosphates (dP and 8-oxo-2'deoxyguanosine (8-oxodG)) to allow introduction of mutations in a highly controlled manner throughout the cDNA. The resultant library of mutants was displayed on the surface of phage and functional polypeptides were selected by several rounds of selection against the cells bearing the receptor for C5a. Following selection only a limited number of residues in C5a were found to be mutated, suggesting that mutations in key residues involved in the maintenance of structure and in receptor binding had been eliminated. The selected C5a sequences had a higher affinity for receptor than wild type phage-C5a conjugates. As this method for analysing the functional characteristics of proteins does not rely on knowledge a priori of structure, it may be useful for affinity maturation or analysis in a wide range of protein ligand/receptor systems. PMID- 11042294 TI - Setting the target for a better cervical screening test: characteristics of a cost-effective test for cervical neoplasia screening. AB - OBJECTIVE: To determine the potential effects on costs and outcomes of changes in sensitivity and specificity with new screening methods for cervical cancer. METHODS: Using a Markov model of the natural history of cervical cancer, we estimated the effects of sensitivity, specificity, and screening frequency on cost-effectiveness. Our estimates of conventional Papanicolaou test sensitivity of 51% and specificity of 97% were obtained from a meta-analysis. We estimated the effect of reducing false-negative rates from 40-90% and increasing false positive rates by up to 20%, independently and jointly. We varied the marginal cost of improving sensitivity from $0 to $15. RESULTS: When specificity was held constant, increasing sensitivity of the Papanicolaou test increased life expectancy and costs. When sensitivity was held constant, decreasing specificity of the Papanicolaou test increased costs, an effect that was more dramatic at more frequent intervals. Decreased specificity had a substantial effect on cost effectiveness estimates of improved Papanicolaou test sensitivity. Most of those effects are related to the cost of evaluation and treatment of low-grade lesions. CONCLUSION: Policies or technologies that increased sensitivity of cervical cytologic screening increased overall costs, even if the cost of the technology was identical to that of conventional Papanicolaou smears. These effects appear to be caused by relatively high prevalence of low-grade lesions and are magnified at frequent screening intervals. Efficient cervical cancer screening requires methods with greater ability to detect lesions that are most likely to become cancerous. PMID- 11042295 TI - Knowledge about human papillomavirus among adolescents. AB - OBJECTIVE: To assess knowledge of human papillomavirus (HPV) among high school aged adolescents. METHODS: We administered written surveys to 523 inner-city high school students in Toronto, Canada, that asked about HPV, other sexually transmitted diseases (STDs), and Papanicolaou testing. We also asked them to report doctor or clinic visits and whether they received sexual health information at those visits. The predictor variables used in analysis were gender and sexual experience. RESULTS: Eighty-seven percent of our population [95% confidence interval (CI) 84%, 89%) had not heard of HPV. Although adolescent women were more knowledgeable about Papanicolaou testing than adolescent men, only 39% of sexually experienced adolescent women knew who should get a Papanicolaou test. Sexually experienced and inexperienced adolescents failed to identify correctly their STD risk. Both genders showed greater knowledge about human immunodeficiency virus (HIV) than other diseases. Among adolescent women, 85% had visited a doctor or clinic within the past year, but only 29% had talked about sexual health. CONCLUSION: Knowledge of HPV infection and cervical cancer screening was low in this urban adolescent population. Improved efforts are needed for prevention of HPV infection and HPV-related cervical changes. Programs modeled after HIV-education programs might be effective. Doctors' offices and clinics providing health care to adolescents should take greater responsibility in sexual health education. PMID- 11042296 TI - Risk factors for cervical stenosis after loop electrocautery excision procedure. AB - OBJECTIVE: To assess frequency of and identify risk factors for the development of cervical stenosis after loop electrosurgical excision procedure. METHODS: We reviewed outpatient charts of women treated by loop excision for cervical dysplasia between August 1996 and January 1998 in the colposcopy clinic at Massachusetts General Hospital. One hundred sixty-four women were evaluated for cervical stenosis during follow-up. Stenosis was considered present if manual dilation was required to allow endocervical sampling with an endocervical currette 3 mm wide. Multivariable analysis with stepwise logistic regression was used to evaluate age, parity, tobacco use, hormonal status, use of oral contraceptives, pathology, previous loop excision, performance of additional endocervical excision, and dimensions of excision specimens as predictors of cervical stenosis. RESULTS: The average age was 32 years. Cervical stenosis occurred in ten of 164 women (6%, 95% CI 3%, 11%). Among factors analyzed, previous loop excision and volume of excision specimen were the only independent predictors of stenosis. CONCLUSION: Cervical stenosis correlated with history of loop excision and volume of tissue removed, suggesting that women who have second excisions or large excisions should be counseled that their risk of stenosis might be higher. PMID- 11042297 TI - Lidocaine spray and outpatient hysteroscopy: randomized placebo-controlled trial. AB - OBJECTIVE: To assess the efficacy of lidocaine spray during outpatient hysteroscopy for reducing procedure-related pain and to identify risk factors for discomfort. METHODS: One hundred twenty-one women were assigned randomly to have application of lidocaine spray or placebo to the uterine cervix during outpatient hysteroscopy. The main outcome measure was pain during hysteroscopy, assessed on a visual analog scale. RESULTS: There was no statistically significant difference between study and control groups in mean age, rate of nulliparity, postmenopausal state, need for cervical dilation, or percentage of women who used hormone replacement therapy. Indications for diagnostic hysteroscopy were similar between groups. Women in the lidocaine group had statistically significantly less pain during the procedure than women in the placebo group (2.2 +/- 1.9 and 3.7 +/- 2.5, respectively; P <.001). Women with abnormal uterine findings (submucous myoma, endometrial polyps, or intrauterine adhesions) had significantly higher pain scores than women with normal cavities (2.2 +/- 1.9 and 3.2 +/- 2.4, respectively; P <.002). Aerosol anesthesia and normal uterine findings were independently associated with less pain. No procedure had to be abandoned because of excessive pain or complications, and no women required hospitalization. CONCLUSION: Women treated with lidocaine spray had significantly less pain. Uterine cavity abnormality might be associated with a higher degree of pain during hysteroscopy. PMID- 11042298 TI - Hormone replacement therapy and glucose metabolism. AB - OBJECTIVE: To determine whether hormone replacement therapy (HRT) alters glucose metabolism. METHODS: Cross-sectional data from the third National Health and Nutrition Examination Survey (1988-1994) included levels of hemoglobin A(1c) in women with diagnosed diabetes and levels of hemoglobin A(1c), fasting and 2-hour glucose, and fasting insulin and C-peptide in women without diagnosed diabetes. We compared mean values for these measures among never, current, and past users of HRT with adjustment for confounders. Types of hormones were not studied. RESULTS: Hormone replacement therapy was used by 8. 6% of diabetic women and 16.7% of women without diagnosed diabetes; 19.3% and 18.5%, respectively, had used HRT in the past. Current use approximately doubled among diabetic women between 1988-1991 and 1991-1994. Current users had lower hemoglobin A(1c) and fasting plasma glucose levels but higher 2-hour glucose levels compared with never and past users. After adjustment for confounding factors, hemoglobin A(1c) levels were 0.1% lower, fasting glucose levels were 3 mg/dL lower, and 2-hour glucose levels were 15 mg/dL higher in current users. Fasting serum insulin and C peptide levels were not associated with HRT use. Duration of HRT use among current users and time since cessation among former users were not associated with measures of glucose metabolism. CONCLUSION: The prevalence of HRT in the United States among diabetic women is approximately half that of women without diabetes diagnoses, although it appears to be increasing. Postmenopausal hormones appear to have no adverse effect on basal glucose metabolism but are associated with slightly elevated postchallenge glucose levels. PMID- 11042299 TI - Criteria for failed labor induction: prospective evaluation of a standardized protocol. AB - OBJECTIVE: To assess the safety and efficacy of a protocol that mandated at least 12 hours of oxytocin administration after membrane rupture before cesarean delivery for failed labor induction in the latent phase. METHODS: Gravidas at or beyond 36 weeks' gestation undergoing indicated induction with cervical dilatation up to 2 cm were studied prospectively. Prior cesarean was an exclusion criterion. If the fetal heart rate pattern was reassuring, cesarean was not permitted before the active phase of labor (4-cm dilatation and at least 90% effacement or 5-cm dilatation regardless of effacement) unless the membranes had been ruptured and oxytocin administered for at least 12 hours. RESULTS: Five hundred nine women were treated according to protocol; 360 (71%) were nulliparas and 149 (29%) were parous. Twenty-five percent of nulliparas and 9% of parous women were delivered by cesarean. After 6 hours of ruptured membranes and oxytocin, 14% of nulliparas were still in the latent phase; 39% of whom delivered vaginally, compared with 7% still in the latent phase after 9 hours (vaginal delivery rate 28%), and 4% after 12 hours (vaginal delivery rate 13%). In contrast, after 6 hours of ruptured membranes and oxytocin, only five (3%) parous women were still in the latent phase. Among those, none remained in the latent phase for 12 hours and all were delivered vaginally. No women had serious complications. Severe neonatal morbidities were infrequent and not related to duration of the latent phase. CONCLUSION: By requiring a minimum of 12 hours of oxytocin after membrane rupture before failed labor induction could be diagnosed, many nulliparas who remained in the latent phase at 6 and 9 hours had safe vaginal deliveries, and failed labor induction was eliminated as an indication for cesarean in parous women. PMID- 11042300 TI - A second-stage partogram. AB - OBJECTIVE: To describe a second-stage partogram based on a system of scoring the descent and position of the fetal head and to use this system for studying progress in the second stage of labor and predicting mode of delivery and obstetric outcome. METHODS: A prospective observational study of 1413 women at term with a singleton, cephalic presentation. The position and station of the fetal head were observed and scored at diagnosis of the second stage of labor, 1 hour later, and then at 30 minute intervals until delivery was achieved. The score at diagnosis of the second stage of labor was assessed for its ability to predict eventual mode of delivery and duration of labor. A normogram was defined for nulliparas and multiparas and was used to define normal and abnormal progress in the second stage, associated factors in the first stage of labor, and mode of delivery. RESULTS: Increasing total score at the start of the second stage of labor is associated with increasing chance of spontaneous vaginal delivery (odds ratio [OR] 1.68 for nulliparas, 1.59 for multiparas), decreasing chance of instrumental vaginal delivery (OR 0.67 for nulliparas, 0.64 for multiparas), and emergency cesarean delivery (OR 0.39 for nulliparas). Abnormal progress as defined by the normogram is associated with use of epidural anesthesia, induction of labor, augmentation, dystocia, and increased incidence of operative delivery. No significant difference is found between normal and abnormal second stages of labor in fetal outcome as determined by Apgar scores. CONCLUSION: The second stage partogram offers an objective basis for management of the second stage of labor. PMID- 11042301 TI - Outpatient cervical ripening with intravaginal misoprostol. AB - OBJECTIVE: To determine if outpatient cervical ripening using misoprostol can initiate labor within 48 hours of medication administration and to determine if time from medication administration to time of delivery is decreased using outpatient cervical ripening. METHODS: Uncomplicated singleton, vertex pregnancies at 41 weeks' gestation or later with Bishop score of 4 or less were eligible for enrollment. Other inclusion criteria included intact membranes, less than eight uterine contractions per hour, a reactive nonstress test, and amniotic fluid index (AFI) over 5 cm. After randomization, 25 micro(cg) of misoprostol or placebo was placed within the posterior vaginal fornix. Patients were continuously monitored for 4 hours, then discharged if not in active labor. Patients returned in 24 hours for a repeat administration of the respective medication. Patients not delivered within 48 hours were admitted for inpatient induction of labor. Statistical analysis was performed with the Fisher, Student t, chi(2), and Mann-Whitney U tests, with P <.05 considered statistically significant. RESULTS: Among the 60 patients enrolled, 27 (45%) received misoprostol and 33 (55%) received placebo. The majority (24 of 27, 88.9%) of study group patients entered active labor within 48 hours after dosing, compared with 16.7% (five of 33) of placebo group patients (P <.001). The time from initial dose to delivery was significantly shorter in the misoprostol group (36.9 +/- 3.8 compared with 61.3 +/- 3.8 hours, P <.001). CONCLUSION: Intravaginal misoprostol is effective for outpatient cervical ripening. No adverse effects were encountered, although further study is required to determine the safety of this treatment regimen. PMID- 11042302 TI - Season and outdoor ambient temperature: effects on birth weight. AB - OBJECTIVE: To determine the extent to which meteorologic factors explain seasonality in birth weight in a developed country. METHODS: Recorded birth weights were collected for all singleton live births after 36 weeks of pregnancy in Northern Ireland between 1971 and 1986. Data on daily maximum and minimum temperatures, rainfall, and hours of bright sunshine were obtained from a local climatologic station for the same period. For each birth, mean daily maximum and minimum temperatures, rainfall, and hours of bright sunshine were calculated for the trimesters of the pregnancy. Linear regression models were constructed with birth weight as the dependent variable and month of birth as a predictor variable. Months of birth were entered in the models as dummy variables. Adjustment was made for year of birth, duration of gestation, maternal age, number of previous pregnancies, sex, and social class of infants at birth and for meteorologic variables relating to each trimester. RESULTS: A clear seasonal pattern in birth weight was observed, with lowest mean birth weight in late spring and summer. Adjusted mean birth weights were 25.5 g, 29.6 g, and 31.6 g lower in May, June, and July, respectively, than in January. This seasonal variation occurred in both sexes, and in female births, it disappeared almost entirely after adjustment for mean daily maximum temperature during the second trimester of pregnancy. CONCLUSION: Infants born during late spring and summer are lighter than those born in winter, which might be the result of exposure to low winter temperatures during midgestation. Pregnant women should keep themselves warm during midpregnancy. PMID- 11042303 TI - Predicting preeclampsia in the second pregnancy from low birth weight in the first pregnancy. AB - OBJECTIVE: To evaluate the effect of low birth weight adjusted for gestational age in first pregnancies on preeclampsia in second pregnancies and to estimate the proportion of preeclampsia in second pregnancies attributable to histories of LBW for gestational age. METHODS: We conducted a cohort study based on linked data from the Medical Birth Registry of Norway, which covered all births in 1967 1992. RESULTS: Women who delivered infants under the third percentile birth weight were three times more likely to have initial or recurrent preeclampsia in second pregnancies than those who delivered infants at or above the tenth percentile. After adjusting for maternal age, year of birth, interpregnancy interval, education, chronic hypertension, diabetes mellitus, and change of partner, the increased risk persisted. Birth weight below the tenth percentile in the first delivery accounted for 10% of the total cases of preeclampsia in the second pregnancy and 30% of recurrent cases. CONCLUSION: A history of low birth weight adjusted for gestational age is associated significantly with subsequent occurrence as well as recurrence of preeclampsia. These findings are consistent with the hypothesis of a shared etiologic factor or recurrent pathophysiologic mechanism for preeclampsia and fetal growth restriction. A history of fetal smallness for gestational age is found in a substantial proportion of all cases of preeclampsia and thus seems to be important in the etiology of preeclampsia. PMID- 11042304 TI - Maternal age and malformations in singleton births. AB - OBJECTIVE: To examine the effect of maternal age on incidence of nonchromosomal fetal malformations. METHODS: Malformations detected at birth or in the newborn nursery were catalogued prospectively for 102,728 pregnancies, including abortions, stillbirths, and live births, from January 1, 1988 to December 31, 1994. Maternal age was divided into seven epochs. Relative risks (RRs) were used to compare demographic variables and specific malformations. The Mantel-Haenszel chi(2) statistic was used to compare age-specific anomalies. Multiple logistic regression analysis was used to adjust for parity. RESULTS: Abnormal karyotypes were significantly more frequent in older women. After excluding infants with chromosomal abnormalities, the incidence of structurally malformed infants also was increased significantly and progressively in women 25 years of age or older. The additional age-related risk of nonchromosomal malformations was approximately 1% in women 35 years of age or older. The odds ratio for cardiac defects was 3.95 in infants of women 40 years of age or older (95% CI 1.70, 9.17) compared with women aged 20-24 years. The risks of clubfoot and diaphragmatic hernia also increased as maternal age increased. CONCLUSION: Advanced maternal age beyond 25 years was associated with significantly increased risk of fetuses having congenital malformations not caused by aneuploidy. PMID- 11042305 TI - Cord leptin level and fetal macrosomia. AB - OBJECTIVE: To determine the relationships among serum leptin, insulin-like growth factor-I, and insulin levels in large for gestational age (LGA) infants. METHODS: Serum samples were collected from maternal veins and umbilical arteries of 52 consecutive, term, LGA neonates of nondiabetic mothers. Maternal and neonatal serum samples were analyzed for levels of leptin, insulin-like growth factor-I, and insulin by specific radioimmunoassays. Multiple regression analysis was used to determine independent risk factors for fetal macrosomia. RESULTS: The independent risk factor significantly associated with fetal macrosomia was umbilical cord leptin concentration (P <.01, beta = 0.59). There was a statistically significant correlation between umbilical cord leptin and insulin like growth factor-I levels and birth weight (r = 0.51, P <.01; r = 0.37, P <.01; respectively). The correlation between umbilical cord insulin levels and birth weight was not statistically significant (r = 0.06, P =.63), nor was that between maternal body mass index and birth weight (r = 0.09, P =.50). CONCLUSION: Our data showed that umbilical cord leptin concentration was an independent risk factor for fetal macrosomia. PMID- 11042306 TI - Outcome of twin gestations after first trimester chorionic villus sampling. AB - OBJECTIVE: To tabulate genetic results and obstetric outcomes of twin pregnancies after first-trimester chorionic villus sampling (CVS). METHODS: The study included 262 consecutive women with twin pregnancies who had first-trimester CVS between 1988 and 1998. RESULTS: Major indications for prenatal diagnosis included maternal age (n = 82), pregnancies after intracytoplasmic sperm injection (n = 114), or both (n = 33). Among 524 fetuses, 519 were sampled adequately. Cytogenetic results were incorrect because of sampling the same fetus twice in two pregnancies. In three pregnancies, contamination caused by mixed sampling made cytogenetic results uncertain. Correct genetic diagnoses were obtained in 509 fetuses, 24 of which had chromosomal abnormalities on direct preparations and four of which had monogenetic conditions. Additional invasive procedures were done on five occasions. Fifteen fetuses were terminated selectively. The total fetal loss rate was 5.5% (28 of 509). The indication for the procedure did not significantly determine the fetal loss rate. The mean +/- standard deviation (SD) gestational age at birth was 35.9 +/- 2.9 weeks, and the mean +/- SD birth weights for twins A and B were 2429 +/- 589.1 g and 2378 +/- 588.5 g, respectively. CONCLUSION: First-trimester CVS is an accurate means of prenatal genetic diagnosis in twins, offering early selective termination in cases of abnormal genetic results in one of the fetuses. PMID- 11042307 TI - Vascular endothelial growth factor and prognosis of cervical carcinoma. AB - OBJECTIVE: To evaluate vascular endothelial growth factor (VEGF) as a marker for predicting lymph node metastasis and an independent prognostic factor of early stage cervical carcinoma. METHODS: One hundred thirty-five women with stage IB IIA cervical carcinoma had radical abdominal hysterectomies and pelvic lymph node dissections. Intratumoral cytosol VEGF concentrations were assayed with enzyme immunoassay. Histopathologic items and cytosol VEGF-influencing clinical outcomes were compared. RESULTS: Twenty-two women (16.3%) who had disease recurrence had higher levels of cytosol VEGF (1020 versus 112 pg/mg protein, P <.001) than those without recurrence. Using a cutoff value of 400 pg/mg protein resulted in best sensitivity of 75%, best specificity of 70%, positive predictive value of 41%, and negative predictive value of 92%. Only overexpressed cytosol VEGF (hazard ratio 6.44, P <.001) was an independent prognostic factor of disease-free survival. The overexpressed cytosol VEGF (hazard ratio 4.50, P =.021) and positive lymphovascular emboli (hazard ratio 4.11, P =.045) were independent prognostic factor of overall survival. CONCLUSION: Cytosol VEGF might be a biomarker for the status of pelvic lymph nodes in early-stage cervical carcinoma and an independent prognostic indicator of its outcome. PMID- 11042308 TI - Radical hysterectomy for cervical cancer in obese women. AB - OBJECTIVE: To estimate the morbidity, adequacy of surgery, and survival of obese women undergoing radical hysterectomy and pelvic lymphadenectomy. METHODS: Patients with stage I and IIa cervical cancer and a body mass index (BMI) over 30 kg/m(2) and absolute weight greater than 85 kg explored with the intent for radical hysterectomy between 1986 and 1998 were identified. Patient characteristics, surgical, pathologic, and follow-up data were extracted and survival curves were generated. RESULTS: Forty-eight obese women were identified who were explored for radical hysterectomy and pelvic lymph node dissection. The median BMI was 36 kg/m(2), and the median weight was 95 kg. Thirty-five patients (73%) had stage Ib1 disease. Despite the obesity of the study group, none had severe comorbidity. The procedure was completed in 46 patients, and abandoned in two because of metastatic disease. For patients undergoing radical hysterectomy and pelvic lymph node dissection, median blood loss was 800 mL. No patient developed fistulas. Residual tumor was present in 26 (57%) hysterectomy specimens, and margins were without disease in 45 specimens (98%). A median of 26 pelvic lymph nodes were obtained per procedure, and six patients (13%) had positive nodes. Five-year overall and disease-free survival are 84% (95% confidence interval [CI] 70.9, 97.5) and 80% (95% CI 65.2, 93.8), respectively, at a median follow-up of 36 months. CONCLUSION: In this carefully selected obese group, we demonstrate that radical hysterectomy and pelvic lymph node dissection can be performed with adequate surgical resection, acceptable morbidity, and excellent survival. PMID- 11042309 TI - Doppler and M-mode ultrasonography to time fetal atrial and ventricular contractions. AB - OBJECTIVE: To compare ease of recording and reliability of ultrasonographic approaches used to time fetal heart atrial and ventricular contractions. METHODS: Seventeen consecutive fetuses seen at our fetal cardiology unit for possible fetal cardiac arrhythmia were included in this study. The same ultrasonographer obtained M-mode tracings of atrial and ventricular free walls, atrial wall and opening of the aortic valves, a peak of the mitral valve, and the opening of the aortic valves; and Doppler signals of flow-velocity waveforms in the outflow tract of the left ventricle and simultaneous flow-velocity waveforms in the aorta and superior vena cava. The outcome measures were rate of successful attempts and intra- and interobserver reliability coefficients. RESULTS: Valid recordings were made for all patients with one M-mode (atrial and ventricular free walls) and two Doppler (intraventricular, superior vena cava, and ascending aorta) approaches. Atrioventricular intervals were significantly longer with M-mode compared with Doppler ultrasonography. Reliability coefficients were excellent (at least 0.89) for all intraobserver measurements. Comparisons of atrioventricular and ventriculoatrial interval measurements made by two observers gave the following intraclass correlation coefficients (95% confidence interval): atrioventricular = M-mode: 0.87 (0.79, 0. 91), left ventricular outflow: 0.93 (0.89, 0.96), superior vena cava-aorta: 0.98 (0.97, 0.99); ventriculoatrial = M-mode: 0.79 (0.67, 0.87), left ventricular outflow: 0.97 (0.95, 0.98); superior vena cava-aorta: 0.99 (0.98, 0.99). CONCLUSION: Fetal atrioventricular intervals measured indirectly from M-mode or Doppler tracings were equally reliable when measured by the same observer; the Doppler approaches had better correlation between measurements made by two different observers. PMID- 11042310 TI - Amniotic fluid index and single deepest pocket: weak indicators of abnormal amniotic volumes. AB - OBJECTIVE: To compare amniotic fluid index (AFI) with the single deepest pocket in the identification of actual abnormal amniotic fluid (AF) volumes. METHODS: One hundred seventy-nine women with singleton pregnancies at the University of Mississippi between March 1994 and June 1999 had ultrasound estimations of AF volume sequentially using the AFI and single deepest pocket techniques. Each woman subsequently had ultrasound-directed amniocentesis with dye-dilution and spectrophotometric calculation of actual AF volume. RESULTS: Actual AF volumes were low (under 5% by volume for gestational age) in 62 women, normal (5-95%) in 100 women, and high (more than 95%) in 17 women. An AFI up to 5 cm (sensitivity 10%, specificity 96%) and a single deepest pocket up to 2 cm (sensitivity 5%, specificity 98%) were similarly inadequate in identifying dye-determined low AF volumes. Likewise, AFI above 20 (sensitivity 29%, specificity 97%) and a single deepest pocket above 8 cm (sensitivity 29%, specificity 94%) were poor in identifying dye-determined abnormally high volumes. CONCLUSION: There was no difference between AFI and single deepest pocket techniques for identifying truly abnormal AF volumes. Both techniques were unreliable for identifying true AF volumes. PMID- 11042311 TI - High and low hemoglobin levels during pregnancy: differential risks for preterm birth and small for gestational age. AB - OBJECTIVE: To examine the association of maternal hemoglobin during pregnancy with preterm birth and small for gestational age (SGA). METHODS: We performed a retrospective cohort analysis of hemoglobin and birth outcome among 173,031 pregnant women who attended publicly funded health programs in ten states and delivered a liveborn infant at 26-42 weeks' gestation. We defined preterm as less than 37 weeks' gestation and SGA as less than the tenth percentile of a US fetal growth reference. RESULTS: Risk of preterm birth was increased in women with low hemoglobin level in the first and second trimester. The odds ratio (OR) for preterm birth with moderate-to-severe anemia during the first trimester (more than three standard deviations [SD] below reference median hemoglobin, equivalent to less than 95 g/L at 12 weeks' gestation) was 1.68 (95% confidence interval [CI] 1.29, 2. 21). Anemia was not associated with SGA. High hemoglobin level during the first and second trimester was associated with SGA but not preterm birth. The ORs for SGA in women with very high hemoglobin level during the first and second trimester (more than three SDs above reference median hemoglobin, equivalent to greater than 149 g/L at 12 weeks' gestation and greater than 144 g/L at 18 weeks') were 1.27 (95% CI 1.02, 1.58) and 1.79 (95% CI 1.49, 2.15), respectively. CONCLUSION: These data highlight the importance of considering anemia and high hemoglobin level as indicators for adverse pregnancy outcome. An elevated hemoglobin level (greater than 144 g/L) is an indicator for possible pregnancy complications associated with poor plasma volume expansion, and should not be mistaken for good iron status. PMID- 11042312 TI - Occipitoposterior position: associated factors and obstetric outcome in nulliparas. AB - OBJECTIVE: To determine factors associated with term delivery in the occipitoposterior position and examine obstetric outcomes from that delivery position in nulliparas. METHODS: We did a retrospective analysis of population based data of 16,781 nulliparas who delivered at term (37-42 weeks) with singleton, cephalic presentations. Factors examined for possible association with occipitoposterior position were fetal weight, maternal age, completed weeks of gestation, epidural analgesia in labor, labor induction, and oxytocin augmentation. Obstetric outcome measures were mode of delivery and percentage of infants with Apgar scores less than 8 at 5 minutes. RESULTS: The frequency of occipitoposterior position was 4. 6%. Fetal weight, epidural analgesia, and oxytocin augmentation were strongly associated with delivery in the occipitoposterior position (odds ratios 1.18, 2.21, 1.44, respectively, P <.001, logistic regression). There was a higher incidence of instrument and emergency cesarean deliveries in occipitoposterior compared with occipitoanterior labors (43.7% versus 24.4%, 41.7% versus 13.7%, respectively, P <.001, the chi(2) test). There was no significant difference in percentage of infants with low Apgar scores at 5 minutes between those who delivered occipitoposterior or occipitoanterior. CONCLUSION: Epidural analgesia and oxytocin augmentation are associated with increased incidence of occipitoposterior position, which leads to increased operative obstetric intervention for delivery. PMID- 11042313 TI - Symptomatic nephrolithiasis complicating pregnancy. AB - OBJECTIVE: To review our experiences with diagnosis and management of symptomatic nephrolithiasis complicating pregnancy and to ascertain the efficacy of renal sonography for initial diagnosis compared with plain x-rays or single-shot intravenous pyelography. METHODS: Perinatal outcomes were evaluated for all pregnant women admitted to Parkland Hospital for nephrolithiasis from 1986 to 1999. Diagnostic studies and management of nephrolithiasis were also evaluated. RESULTS: During the 13-year study period, 57 pregnant women had 73 admissions for symptomatic nephrolithiasis. Symptomatic nephrolithiasis complicated 1 in 3300 (0.03%) deliveries at our institution. Only 12 women (20%) had a history of renal calculi. Mean gestational age at diagnosis was 23 weeks. Imaging techniques included renal ultrasonography, plain abdominal x-ray, and single-shot intravenous pyelography. Calculi were visualized in 21 of 35 (60%) renal ultrasonographic examinations and 4 of 7 (57%) abdominal x-ray studies when these were performed as the initial test. In contrast, urolithiasis was discovered in 13 of 14 (93%) instances in which intravenous pyelography was performed as the initial diagnostic test. When sonography was negative (n = 14), renal calculi were confirmed by single-shot intravenous pyelography (n = 8). Although 43 of 57 (75%) of symptomatic episodes responded to conservative management, 10 women required ureteral stents, 3 needed percutaneous nephrostomy tubes, and 2 underwent ureteral laser lithotripsy for resolution. CONCLUSION: Although the convenience and safety of ultrasonography to initially diagnose nephrolithiasis are indisputable, 40% of calculi were missed when this method alone was used. Thus, if nephrolithiasis is still suspected clinically despite ultrasonographic findings, single-shot pyelography is recommended. PMID- 11042314 TI - Antepartum or postpartum isoniazid treatment of latent tuberculosis infection. AB - OBJECTIVE: To compare health outcomes and costs of different strategies for treatment of latent tuberculosis infection in pregnancy. METHODS: Using a Markov decision-analysis model, the following three strategies were evaluated for treatment of latent tuberculosis infection in pregnancy, defined as positive tuberculin skin reaction of 10 mm or greater and negative chest radiograph: no treatment, antepartum isoniazid administration, in which women were given 300 mg of isoniazid with pyridoxine beginning at 20 weeks' gestation for 6 months; and postpartum isoniazid, in which women were given isoniazid and pyridoxine for 6 months after delivery. Sensitivity analyses were performed for a wide range of probability and cost estimates, and considered discount rates. RESULTS: Under base-case assumptions, the fewest cases of tuberculosis within the cohort occurred with antepartum treatment (1400 per 100,000) compared with no treatment (3300 per 100,000) or postpartum treatment (1800 per 100,000). Antepartum treatment resulted in a marginal increase in life expectancy due to the prevented cases of tuberculosis, despite more cases of isoniazid-related hepatitis and deaths, compared with no treatment or postpartum treatment. Antepartum treatment was the least expensive. Only if the case-fatality rate for tuberculosis was tenfold lower than the base-case and the risk of fatal hepatitis tenfold higher did antepartum treatment become the least advantageous strategy. CONCLUSION: Rather than delaying treatment until postpartum, consideration for antepartum treatment of latent tuberculosis during pregnancy should be given. If isoniazid is not administered antepartum, then efforts to improve postpartum compliance should be instituted, as either antepartum or postpartum treatment is better than no treatment. PMID- 11042315 TI - Open laparoscopy: 29-year experience. AB - OBJECTIVE: To describe the safety and efficacy of open laparoscopy as a method of access to the abdominal cavity for laparoscopic surgery. METHODS: We reviewed retrospectively all cases of open laparoscopy we did between August 1970 and June 1999. RESULTS: Twenty-seven (0.5%) of 5284 patients who had open laparoscopies during the study years developed complications related to primary access. Twenty one had minor wound infections, four had minor hematomas, one developed an umbilical hernia that required reoperation, and one had an inadvertent injury to the small bowel that was repaired intraoperatively without adverse outcome. Access to the abdominal cavity was generally secured in 3-10 minutes. CONCLUSION: Open laparoscopy was associated with no method failure or life-threatening complications. Minor and medium risk complications occurred at a rate of 0.5%. Open laparoscopy is a safe, effective method of accessing the abdominal cavity. PMID- 11042316 TI - Combined tobacco and alcohol use by pregnant and reproductive-aged women in the United States. AB - OBJECTIVE: To assess trends in the concurrent use of alcohol and tobacco among pregnant women. METHODS: Using behavioral risk factor surveillance system data from 1987 through 1997, we determined the prevalence of concurrent tobacco and alcohol use among women aged 18-44 years by pregnancy status and indirectly estimated pregnancy-related disuse rates. RESULTS: The percentage of women who used alcohol and tobacco decreased significantly from 1987 to 1990 among pregnant (5.4% to 3.0%) and nonpregnant women (17.6% to 14.2%), but thereafter did not change significantly. The estimated pregnancy-related disuse rate of tobacco and alcohol increased insignificantly from 70% in 1987 to 82% in 1997. Among women who used both substances, pregnancy-related disuse was slightly greater for alcohol alone (74%) than for tobacco alone (52%). There was not a significant decline in concurrent use of tobacco and alcohol between 1987 and 1997 among women 18-20 years old (pregnant, 4.4% to 3.6%; nonpregnant, 13.5% to 13.7%). That age group also showed a smaller pregnancy-related disuse rate than older women (1997, 74% versus 83%). CONCLUSION: The steady trend in concurrent use of tobacco and alcohol by young women emphasizes the need for enhanced efforts to reduce the initiation of tobacco and alcohol use by young people. Women who report abuse of tobacco or alcohol should be evaluated for abuse of both substances, and interventions should address abuse of both substances. PMID- 11042317 TI - Post-term induction of labor revisited. AB - Post-term pregnancy (longer than 42 weeks or 294 days) occurs in approximately 10% of all singleton gestations. The adverse outcomes of post-term pregnancy include a substantial increase in perinatal mortality and morbidity. ACOG currently recommends induction of labor for low-risk pregnancy during the 43rd week of gestation. However, that recommendation dates from 1989. Recent reports mandate reconsideration of the management of post-term pregnancy, including reinterpretation of the statistical risk of stillbirth in post-term pregnancies using ongoing (undelivered) rather than delivered pregnancies as the denominator, which shows a far higher risk to post-term fetuses than believed. Recent data also suggest that the risk of cesarean delivery after induction of labor at term is lower than reported, possibly because of improvements in methods for cervical ripening. Those findings provide rationale for earlier labor induction in low risk pregnancies. PMID- 11042318 TI - Does full-term pregnancy at a young age protect women against breast cancer through hCG? AB - Recent studies found that human and animal breast tissues and human breast cell lines contain low levels of receptors that bind hCG and its structural and functional homologue, LH. Those gonadotropins exert numerous anticancer effects in breast cancer models and cells, which might explain decreased breast cancer incidence in women who complete full-term pregnancies at a young age. The new findings also imply that premature chronic elevations of LH levels might contribute to decreased breast cancer incidence in women with early menopause, and elevated LH levels might contribute to a better prognosis after ovariectomy. Those findings predict that breast cancer risk might be reduced by early hCG treatment of women who plan to delay their first pregnancies; prophylactic hCG treatment might help women with family histories of breast cancer or oncogene mutations that predict breast cancer; and better prognoses might result when hCG is administered to breast cancer tissue. PMID- 11042320 TI - Hand-held computer operating system program for collection of resident experience data. AB - OBJECTIVE: To describe a system for recording resident experience involving hand held computers with the Palm Operating System (3 Com, Inc., Santa Clara, CA). PROGRAM DESCRIPTION: Hand-held personal computers (PCs) are popular, easy to use, inexpensive, portable, and can share data among other operating systems. Residents in our program carry individual hand-held database computers to record Residency Review Committee (RRC) reportable patient encounters. Each resident's data is transferred to a single central relational database compatible with Microsoft Access (Microsoft Corporation, Redmond, WA). Patient data entry and subsequent transfer to a central database is accomplished with commercially available software that requires minimal computer expertise to implement and maintain. The central database can then be used for statistical analysis or to create required RRC resident experience reports. As a result, the data collection and transfer process takes less time for residents and program director alike, than paper-based or central computer-based systems. CONCLUSION: The system of collecting resident encounter data using hand-held computers with the Palm Operating System is easy to use, relatively inexpensive, accurate, and secure. The user-friendly system provides prompt, complete, and accurate data, enhancing the education of residents while facilitating the job of the program director. PMID- 11042319 TI - Rethinking maternal-fetal conflict: gender and equality in perinatal ethics. AB - Practitioners who care for pregnant women face dilemmas when their patients use illicit drugs, reject medical recommendations, or cause fetal harm. Many ethics scholars characterize those situations as maternal-fetal conflicts. In conflict based models, maternal rights are considered to conflict with fetal rights, or moral obligations owed to pregnant women are considered to conflict with those owed to their fetuses. I offer an alternative model of pregnancy ethics by applying relational and equality-based moral theories to situations of fetal harm by pregnant women. In this model, clinicians faced with ethical dilemmas should attempt to understand pregnant women and their decisions within their broad social networks and communities, ask how the clinician's personal standpoint influences outcomes judged to be ethical, and determine whether the clinician's ethical formulations reduce or enhance existing gender, class, or racial inequality. This model focuses on the mutual needs of pregnant women and fetuses rather than on their mutually exclusive needs. It also avoids many pitfalls of traditional ethical formulations, specifically their tendency to neglect gender specific modes of moral reasoning, their implicit assumptions that application of universal principles like autonomy and beneficence results in objective ethical solutions, and their failure to account for the ways that projecting fetal needs perpetuates social inequalities. This model provides the ethical foundations for moving law and policy away from criminalization and toward prevention of prenatal harm. PMID- 11042321 TI - The adventure of the three abnormal paps. PMID- 11042322 TI - The bloodless blood knot. AB - Just as it is difficult to describe adequately the exhilaration one feels when using a fly rod to land a trout caught from a mountain stream, there is also a tremendous amount of satisfaction in the successful completion of an obstetric operation. Until recently, we were woefully ignorant of how fly fishing expertise could benefit pregnancy. We report with great pride an instance in which fly fishing knot skill was essential to successful placement of a cervical cerclage for a woman with an incompetent cervix. PMID- 11042323 TI - Morphological and ultrastructural studies of some acritarchs from the Lower Cambrian Lukati Formation, Estonia. AB - Six acritarch species from the Lukati Formation were studied using a combination of techniques, including transmitted light, scanning electron (SEM) and transmission electron (TEM) microscopy. New details of wall ultrastructure, surface microsculpture and internal morphology of the vesicle and processes significantly add to the previously known morphological features and increase the understanding of the form-genera Archaeodiscina, Globosphaeridium, Comasphaeridium, Skiagia, Tasmanites and Leiosphaeridia. Examination of microfossils using TEM revealed a substantial variation in wall ultrastructure among acritarchs. The diversity includes four structural types of vesicle wall in addition to their single- and multi-layered structure and the variable thickness of the wall. These are: electron-tenuous and fibrous; electron-dense and homogeneous; electron-dense and homogeneous but perforated by radial canals; and composite laminated structure. Morphologically recognised groupings of acritarchs (acanthomorphic, disphaeromorphic, sphaeromorphic) and tasmanitid taxa appear to be characterised by particular features of the wall structure, although the wall structure in itself may not be directly indicative of systematic relationships. Structurally diverse vesicle walls are observed in Tasmanites and Leiosphaeridia, taxa that both have been interpreted, based on other lines of evidence, to be of prasinophycean (green algal) affinities. The distinct wall ultrastructure of the Leiosphaeridia studied is similar to that of extant green algal genera, which provides evidence that some Cambrian leiosphaerids were chlorophycean algae, probably related to the Order Chlorococcales. Previous research and interpretations of the wall ultrastructure are also briefly discussed. PMID- 11042324 TI - The Ordovician acritarch genera Tranvikium and Ampullula: their relationship and taxonomy. AB - A restudy of the Ordovician (Arenig-Llanvirn) acritarch taxa Tranvikium polygonale Tynni, 1982, and Ampullula suetica Righi, 1991, indicates that they represent extremes in a single morphological plexus. At one extreme are forms with a polar 'excystment' aperture (closed by an operculum or two opercular pieces) and a smaller opening (plugged or open) at the opposite pole; at the other are forms lacking a polar aperture but having, at the opposite pole, a tube open distally and plugged or open basally. New morphological terms for these structures are proposed. The genera and species are treated as synonyms and an emended diagnosis is given for Tranvikium polygonale, incorporating this whole morphological plexus. The possible purposes of the structures exhibited and the likely affinity of T. polygonale to various groups of algae are discussed. The emendation of Ampullula by Brocke (1997) is rejected and the genus Stelomorpha Yin, 1994 retained, with an emended diagnosis. The new combination Stelomorpha princeps (Brocke, 1997) Uutela and Sarjeant is proposed. PMID- 11042325 TI - Taxonomy and palaeoecology of dinoflagellate cysts from Upper Oligocene freshwater sediments of Lake Enspel, Westerwald area, Germany. AB - Freshwater dinoflagellates play an important role as primary producers in the lacustrine environment. A new species of dinoflagellates, Cleistosphaeridium lacustre, is described from Upper Oligocene sediments of palaeo-lake Enspel. They are associated with other phytoplankton, such as diatoms, chrysophytes, green algae and benthic cyanobacteria. Mass occurrences of this species are interpreted as algal blooms and may partly reflect seasonal successions. This phenomenon was controlled by volcanic activities in the depositional area, which led to an increase in nutrient supply. PMID- 11042326 TI - Frenelopsis alata and its microsporangiate and ovuliferous reproductive structures from the Cenomanian of Bohemia (Czech Republic, Central Europe). AB - The conifer, Frenelopsis alata (K. Feistmantel) E. Knobloch (Cheirolepidiaceae), occurring mostly in the Cenomanian of Europe, is revised on the basis of the type material. Its comparison with relevant species of Frenelopsis is discussed.The ovuliferous cone associated with the genus Frenelopsis is recorded for the first time. For the associated ovuliferous cones of Frenelopsis, a new genus, Alvinia, is introduced in a new combination for the type: Alvinia bohemica (Velenovsky) comb. n. Its association with Frenelopsis alata is based on the presence of Classopollis pollen adhering to ovuliferous cone scales, and the same type of pollen found in the microsporangiate cone of F. alata, the same cuticle pattern present on ovuliferous cones, sterile twigs and microsporangiate cones of F. alata, and also the co-occurrence of ovuliferous cones or their scales and sterile twigs of F. alata.Large ovuliferous cones of Alvinia bohemica are formed by helically arranged ovuliferous scales subtended by bracts. Each ovuliferous cone scale displays one or two seeds covered by a covering flap, and three appendages, which form distally a funnel-like structure lined in its inner part by long trichomes. Numerous pollen grains of Classopollis adhere to the trichomes, and the structure is considered to function as a protostigmatic area.The ovuliferous cones of Alvinia differ from similar cones of the Cheirolepidiaceae, Hirmeriella and Tomaxellia, mainly in a high state of unification of the ovuliferous cone scale, reduction of appendages and in a presence of the protostigmatic funnel-like structure.The ovuliferous cones, Alvinia bohemica, rarely occur intact, so it is assumed that they disintegrate when mature. It seems likely that they were not woody. This assumption is supported by the flattened appearance of cones and their cone scales in the sediment, their flexibility and the absence of massive coaly matter known from cones of the Taxodiaceae and Cupressaceae. It is proposed that this type of ovuliferous cone scale indicates a specialized type of pollination. In addition, it is suggested that cone scales enclosing seeds play an important role in propagation. PMID- 11042327 TI - Use of plots to define pollen-vegetation relationships in densely forested ecosystems of Tropical Africa. AB - Modern soil samples from South Congo were analyzed for pollen content and compared to forest inventories to define modern pollen-vegetation relationships. A correspondence analysis (CA) was applied independently to botanical and pollen data and a hierarchical cluster analysis to pollen data only. Subsequently, a CA using a presence-absence approach has been made to directly compare the two types of data. Results show that the pollen rain and floristic composition of the sampled sites are not directly linked to altitudinal or precipitation gradients, but clear evidence of variation in relation to hygromorphy and soil type is detected. The forests occurring in swampy environments are well differentiated from the forests developed on well-drained soils by pollen and floristic data. Among forests on well-drained soils, a good distinction can be made between those growing on sandy soils and those growing on ferralitic soils. The comparison between pollen spectra and vegetation shows site-to-site variations in pollen assemblages in relation to the floristic heterogeneity of forests, and it appears that few taxa show a good correlation between plant cover and pollen abundance. PMID- 11042328 TI - Spatial variation of modern pollen in Oregon and southern Washington, USA. AB - Surface sediments from 95 lakes provide information on the spatial variation of modern pollen spectra in Oregon and southern Washington. Percentages for 13 pollen types were compared within and between vegetation zones to characterize regional patterns of pollen spectra. The percentage data were also compared with climate variables to determine relationships between pollen percentages and regional climate gradients. The composition of modern pollen spectra corresponds well with the distribution of the pollen producers. Most pollen assemblages were generally dominated by Pinus, but those west of the Cascade Range were dominated by Alnus. Low percentages of Pseudotsuga/Larix, Tsuga mertensiana, Abies, and Picea pollen coincided with local occurrence of the trees. The distributions of the pollen data were arranged along gradients of temperature and effective moisture. West of the Cascade Range, Alnus, Tsuga heterophylla, Pseudotsuga/Larix, and Cupressaceae pollen were abundant and correlate well with moderate temperature and high effective moisture. In the shrub-steppe and woodlands east of the Cascade Range, where effective moisture is low, Artemisia, Cupressaceae, and Pinus pollen were dominant. At high elevations, Pinus, T. mertensiana, Abies, and Picea were common pollen types in areas with short growing seasons and high effective moisture. Pollen percentages collected from lake surface sediments, moss polsters, and soils were compared within a number of vegetation types to assess their similarity. The three types of sample yielded similar results for forested areas, but lake sediment samples from upper- and lower-treeline sites captured a more regional picture of the vegetation. PMID- 11042329 TI - A reconsideration of Wattieza Stockmans (here attributed to Cladoxylopsida) based on a new species from the Devonian of Venezuela. AB - A single small plant fragment from the Middle Devonian of Belgium, first illustrated by Stockmans as Wattieza givetiana, is redescribed. It consists of a small central axis bearing on it whorls of four several-times bifurcate laterals that are terminally recurved. One tip is here shown to have attached one elongate ellipsoid sporangium. More abundant Middle Devonian fossils from Venezuela have similar characteristics, although they are more robust. They, together with more simple sterile examples, are demonstrated to be attached laterally and irregularly to large axes that are themselves found distal to a digitate division of a substantial branch with an enlarged base. All branches, axes and the central axes of the lateral branching systems are covered in small tangentially elongate depressions. These characteristics, taken together, demonstrate that Wattieza is a large plant of similar habit to Pseudosporochnus Potonie et Bernard. Wattieza joins Lorophyton Fairon-Demaret et Li, Calamophyton Krausel et Weyland and Pseudosporochnus as morphologically well-circumscribed Devonian members of the phylogenetically important plant group Cladoxylopsida. Wattieza casasii Berry sp. nov. is erected to accommodate the Venezuelan material. PMID- 11042330 TI - Visan miospore biostratigraphy and correlation of the Poti Formation (Parnaba Basin, northern Brazil). AB - The Poti Formation, which consists mainly of sandstones with minor proportions of carbonaceous shales and other siliciclastic lithologies, represents all the Visean strata thus far recorded in the Parnaiba Basin, northern Brazil.Well preserved miospores featuring species with both Southern Euramerican and Gondwanan affinities have been recovered from this formation in four well sections. The most characteristic species are listed in this paper, and brief systematic descriptions are presented for the most significant species, along with comments on their biostratigraphy. A new generic combination is proposed: Cordylosporites magnidictyus (Playford and Helby) Loboziak and Melo comb. nov. Comparisons with miospores illustrated from the Grand Erg Occidental, Algerian Sahara, are tentatively proposed.In terms of the Western European Carboniferous palynozonation, miospore assemblages from the Poti Formation are assignable to the Perotrilites tessellatus-Schulzospora campyloptera (TC)-Raistrickia nigra Triquitrites marginatus (NM) zonal range. This corresponds to the upper part of the Holkerian and the whole Asbian, which are British regional stages for the lower to middle parts of the upper Visean. The Visean age formerly attributed to biostratigraphic interval XII of Petrobras' regional palynostratigraphic scheme is therefore confirmed. As already noted in our recent investigations of the Faro Formation in the Amazon Basin and equivalent strata of the Solimoes Basin, latest Tournaisian and early to middle Visean sections are either absent or barren of characteristic miospores in the Parnaiba Basin as well. PMID- 11042331 TI - New taxa of angiosperm pollen, miospores and associated palynomorphs from the early Late Cretaceous of Egypt (Maghrabi Formation, Kharga Oasis). AB - A palynological investigation of samples from various boreholes in the Maghrabi Formation (Kharga Oasis, southern Egypt) resulted in the recovery of pollen and spore assemblages associated with rare marine palynofossils (dinoflagellates, foraminiferal linings) and freshwater algae (e.g. Botryococcus, Ovoidites parvus, Pediastrum, Scenedesmus). The general composition of the assemblages is largely consistent with the estuarine and tidal flat conditions characteristic of the Maghrabi Formation.The formal descriptions of the following new taxa are given: Cicatricosisporites kedvesii Schrank, sp. nov., Equisetosporites lawalii Schrank, sp. nov., Dettmannaepollenites clavatus Schrank, sp. nov., and Integritetradites porosus Schrank and Mahmoud, gen. nov. and sp. nov. Combined scanning electron microscopic and light microscopic techniques have been applied to hand-picked grains to illustrate the new taxa. The palynological ages assigned to the Maghrabi samples are mainly based on angiosperm pollen and range from undifferentiated Cenomanian for an Integritetradites porosus assemblage without triporates to Late Cenomanian-Early Turonian for another assemblage which has I. porosus associated with rare triporate pollen grains (Proteacidites/'Triorites' spp.). PMID- 11042339 TI - Expression of bcl-2, bax, and caspase-3 in the brain of the developing rat. AB - Naturally occurring neuronal death (NOND) is generally considered to be apoptotic. Apoptosis is an active form of cell death in which the regulation of specific proteins produces anti- or pro-apoptotic signals. Two of the protein families involved in this regulation are the bcl proteins and caspases. A quantitative immunoblotting technique was used to examine the temporal expression of bcl-2, bax, and two isoforms of caspase 3 (an active 20 kDa isoform and the inactive 32 kDa precursor) throughout the developing neuraxis. Long-Evans rat fetuses were collected on gestational day (G) 16 and G19, and pups were harvested on postnatal day (P) 0, P3, P6, P12, P21, and P30. Brains were divided into five segments: cortex, thalamus, midbrain, medulla/pons, and cerebellum. In general, the expression of bax increased and the ratio of bcl-2 expression to bax expression decreased concurrent with published data on the onset of NOND in a given area. The timing of these events was paralleled by an increase in the expression of active caspase 3. Unlike the bcl proteins, caspase 3 expression returned toward fetal levels as the brain matured. The timing of the changes in bcl protein and caspase expression show that both protein families are involved in promoting neuronal death. Reductions in caspase expression (and not bcl-2 and bax expression) are key to ending the period of NOND. PMID- 11042340 TI - Nitric oxide synthase activity and inhibition after neonatal hypoxia ischemia in the mouse brain. AB - Despite the emergence of therapies for hypoxic-ischemic injury to the mature nervous system, there have been no proven efficacious therapies for the developing nervous system. Recent studies have shown that pharmacological blockade of neuronal nitric oxide synthase (nNOS) activity can ameliorate damage after ischemia in the mature rodent. We have previously shown that elimination of nNOS neurons, either by targeted disruption of the gene or by pharmacological depletion with intraparenchymal quisqualate, can decrease injury after hypoxia ischemia. Using a simpler pharmacological approach, we studied the efficacy of a systemically administered NOS inhibitor, 7-nitroindazole, a relatively selective inhibitor of nNOS activity. Using multiple doses and concentrations administered after the insult, we found that there was only a trend for protection with higher doses of the drug. A significant decrease in NOS activity was seen at 18 h and 5 days in the cortex, and at 2 h and 18 h in the hippocampus after the hypoxia ischemia. nNOS expression decreased and remained depressed for at least 18 h after the insult. When nNOS expression was normalized to MAP2 expression, a decrease was seen at 18 h in the cortex and at 2 and 18 h in the hippocampus. These data suggest that further inhibition of NOS activity at early timepoints may not provide substantial benefit. At 5 days after the insult, however, NOS activity and normalized nNOS expression returned to baseline or higher in the hippocampus, the region showing the most damage. These data suggest that delayed administration of nNOS inhibitor after hypoxic-ischemic injury might be beneficial. PMID- 11042341 TI - NGF expression in the developing rat brain: effects of maternal separation. AB - A number of studies have shown that mothering style in rodents can produce neuroendocrine, neurochemical and behavioural changes in the adult, although the basic mechanisms initiating this cascade of events still need to be investigated. Long term changes in neuronal function might be due to alterations in the expression of neurotrophins which have been shown to promote neuronal survival, differentiation and function during development, such as Nerve Growth Factor (NGF). NGF is essential for proper development of sympathetic and neural crest derived sensory neurons of the peripheral nervous system as well as of central cholinergic neurons. In previous studies, using a maternal separation paradigm, we have shown that NGF expression is increased in the dentate gyrus and the hilus of the hippocampus as a result of brief (45 min) maternal separations. In the present study neonatal rats were separated for longer periods of time (up to 3 h) and at different ages during development (9 and 16 days postnatally). Results indicate that the effects of maternal separation on NGF expression are stronger with longer separations and are not restricted to the hippocampal region but can be seen also in other brain areas. Overall these results indicate that external factors, such as the presence/absence of the mother, can modify neurotrophic factor's availability in the brain, thus indicating NGF as a potential player in environmentally-mediated brain plasticity during development. PMID- 11042342 TI - Postnatal development of opioid receptors modulating acetylcholine release in hippocampus and septum of the rat. AB - The postnatal development of presynaptic opioid receptors inhibiting the release of acetylcholine (ACh) was studied in rat brain hippocampus, medial septum (MS) and diagonal band of Broca (DB). To this end, the corresponding brain slices (350 microm thick) of rats of various postnatal ages (postnatal day 4 [P4] to P16, and adult) were preincubated with [(3)H]choline and stimulated twice for 2 min (S(1), S(2): at 3 Hz, 2 ms, 60 mA) during superfusion with physiological buffer containing hemicholinium-3. In parallel, the activity of choline acetyltransferase (ChAT) was determined in crude homogenates of the tissues as a marker for the development of cholinergic neurons. At any postnatal age, the electrically evoked overflow of tritium from slices preincubated with [(3)H]choline was highest in the DB, followed by the MS and the hippocampus. The evoked [(3)H]overflow increased with postnatal age, reached about 50% (MS, DB) or 30% (hippocampus) of the corresponding adult levels at P16 and correlated significantly with the corresponding ChAT activities. Presence of the preferential mu-opioid receptor agonist DAMGO during S(2) significantly inhibited the evoked overflow of tritium already at P4 in DB and MS, whereas in the hippocampus significant inhibitory effects were first observed at P8 only. Moreover, adult levels of inhibition due to DAMGO were reached at P16 in the DB and MS but not in the hippocampus. In septal areas, also the effect of the preferential delta-opioid receptor agonist DPDPE on the evoked [(3)H]overflow was studied: in contrast to DAMGO, however, significant inhibitory effects of DPDPE were first observed at P12 only. In conclusion, the postnatal development of presynaptic mu-opioid receptors on cholinergic neurons in the DB and MS starts earlier than in the hippocampus and precedes that of presynaptic delta-opioid receptors. PMID- 11042343 TI - Developmental differences in alternative splicing of the NR1 protein in rat cortex and cerebellum. AB - Expression of the C-terminal cassettes of the NR1 protein was examined using a quantitative Western blot method with cassette-specific antibodies. Measurements were made of the percent of total NR1 protein that contained a specific cassette in both the cerebellum and cortex over development. In the cortex, the C1 cassette was shown to be present in about half of total NR1 protein with no change over development. While about half of total NR1 in the cerebellum at postnatal day 42 (P42) contained the C1 cassette, little NR1 protein with this cassette was seen at young ages. In both the cerebellum and cortex, the C2 and C2' cassettes showed opposite developmental patterns, with the C2 cassette decreasing and the C2' cassette increasing over age. Together with previous data on the expression of the N1 cassette, this study describes the alternatively spliced forms of NR1 protein that are dominant at different ages. In the young cerebellum, the NR1(001) form appeared dominant, while in the young cortex there appeared to be a mix of NR1(001) and NR1(011). The most common splice forms of NR1 protein in the adult cerebellum appeared to be NR1(111) and NR1(100). In the adult cortex, there appeared to be a mix of NR1(001) and NR1(011). These data on the expression of the alternatively spliced forms of NR1 allow predictions on the possible characteristics of NMDA receptors in different regions at specific ages. PMID- 11042344 TI - Activity-dependent development of spontaneous bioelectric activity in organotypic cultures of rat occipital cortex. AB - The development of spontaneous bioelectric activity (SBA) in organotypic tissue cultures (OTCs) from rat occipital cortex was studied by means of extracellular recording techniques in OTCs grown normally for 6-51 days in vitro (DIV), and in OTCs in which SBA had been silenced from DIV 4 on for 2 to 3 weeks by elevating the Mg(2+) levels in the growth medium. The proportions of spontaneously active neurones increased from about 25% at 6-14 DIV to more than 80% beyond the third week in vitro. Mature neurones discharged at shorter intervals and more vigorously than immature neurones; the developmental increase in firing rate was not significant, however. In OTCs 6-14 DIV the majority of spontaneously active neurones fired sluggishly in a regular manner. The remaining neurones fired action potentials in the form of discrete bursts resembling interictal activity in vivo. The proportions of these neurones increased from about 40% at 6-14 DIV to more than 80% beyond the third week in vitro. During development in vitro the mean burst duration increased from 3.5 s to about 8 s whereas the mean burst rate (between 0.7-1 bursts/min) remained constant. Activity-deprived neurons had low firing rates and fired action potentials in the form of discrete bursts with a mean burst rate of 0.4/min. The proportions of spontaneously active neurons, the variability of neuronal firing and the viability of the explants either were not altered by the activity blockade or had recovered to control values after 5-6 days in normal growth medium. We conclude that in OTCs of rat neocortex the absence of SBA during development in vitro delays the maturation of excitatory mechanisms responsible for the developmental increase in firing intensity. The development of burst firing modes is less affected by activity blockade. PMID- 11042345 TI - Quantification of the alpha(3) subunit of the Na(+)/K(+)-ATPase in developing rat cerebellum. AB - Cerebellar Purkinje neurons of rats have been shown to exhibit a progressive increase in resting membrane potential as the animals develop postnatally. The magnitude of this increase was equivalent in magnitude to the increase in the depolarizing action of ouabain, consistent with a role for the Na(+)/K(+)-pump in the hyperpolarization. Ouabain binding sites in whole cerebellum also increased with age. The present study was undertaken to confirm that the increases in ouabain binding and the electrophysiological responses to ouabain were a consequence of increases in the sodium pump and to determine whether the changes seen at the whole organ level were reflective of changes taking place at the cellular level. Using antibodies directed against the alpha(1), alpha(2), and alpha(3) subunits of the Na(+)/K(+)-ATPase, rats between 13 and 19 days of age exhibited a statistically significant increase in the relative amount of the alpha(3) subunit at the level of the whole organ, as determined by Western and slot blot analyses, with no change in the levels of either the alpha(1) or the alpha(2) subunit. Using immunohistochemistry, the alpha(3) subunit was shown to increase in both the Purkinje cell layer and the white matter during this postnatal time period, while the alpha(1) subunit increased in the granular layer. These results support and extend previous work, which pointed to a role for the electrogenic sodium pump in the developmental increase in Purkinje cell membrane potential. Furthermore, the data provide a cellular mechanism underlying the increase in resting membrane potential, that is, by the specific modulation of the alpha(3) subunit isoform. PMID- 11042346 TI - The role of NF-AT transcription factors in T cell activation and differentiation. AB - The family of genuine NF-AT transcription factors consists of four members (NF AT1 [or NF-ATp], NF-AT2 [or NF-ATc], NF-AT3 and NF-AT4 [or NF-ATx]) which are characterized by a highly conserved DNA binding domain (is designated as Rel similarity domain) and a calcineurin binding domain. The binding of the Ca(2+) dependent phosphatase calcineurin to this region controls the nuclear import and exit of NF-ATs. This review deals (1) with the structure of NF-AT proteins, (2) the DNA binding of NF-AT factors and their interaction with AP-1, (3) NF-AT target genes, (4) signalling pathways leading to NF-AT activation: the role of protein kinases and calcineurin, (5) the nuclear entry and exit of NF-AT factors, (6) transcriptional transactivation by NF-AT factors, (7) the structure and expression of the chromosomal NF-AT2 gene, and (8) NF-AT factors in Th cell differentiation. The experimental data presented and discussed in the review show that NF-AT factors are major players in the control of T cell activation and differentiation and, in all likelihood, also of the cell cycle and apoptosis of T lymphocytes. PMID- 11042347 TI - Phorbol ester synergistically increases interferon regulatory factor-1 and inducible nitric oxide synthase induction in interferon-gamma-treated RAW 264.7 cells. AB - The roles of PKC in iNOS induction by IFN-gamma have been shown in some cell types. The effect of a PKC activator, phorbol ester, in iNOS induction is thought to be due to multiple mechanisms, and it is necessary to examine the involvement of phorbol ester on IFN-gamma-induced iNOS in detail. In the present study, we investigated the mechanisms of phorbol ester on IFN-gamma-induced iNOS in RAW 264.7 cells. PMA synergistically increased iNOS activity, protein and mRNA levels in IFN-gamma-treated RAW 264.7 cells. PMA together with IFN-gamma increased iNOS mRNA without affecting the iNOS mRNA degradation, suggesting that the synergistic effect of PMA on IFN-gamma-induced iNOS mRNA production may depend on the elevation of the transcription rate rather than a prolongation of mRNA stability. The DNA binding proteins that are involved in the regulation of iNOS expression are mainly NF-kappa B and IRF-1. IRF-1 transcriptionally regulates many IFN inducible genes such as iNOS whose promoter contains an IRF-1 binding site. PMA might modulate iNOS induction as a cosignal with IFN-gamma in RAW 264.7 cells because the synergistic effect of PMA was mediated through IRF-1, rather than NF kappa B. Ro 31-8220, a PKC inhibitor, decreased iNOS activity, protein, mRNA levels and IRF-1 activity, indicating that the effect of PMA on iNOS induction might occur via the PKC pathway. It is evidence that PKC plays an important role in IRF-1 activation and that phorbol ester has a synergistic effect on iNOS induction through IRF-1 activation in IFN-gamma-treated RAW 264.7 cells. The synergistic effect of PMA on IFN-gamma-induced IRF-1 binding activity was observed in macrophage cell line J774 cells as well as RAW 264.7 cells, but not in thioglycollate-elicited peritoneal macrophages. PMID- 11042348 TI - Evidence for the presence of cGMP-dependent protein kinase-II in human distal colon and in T84, the colonic cell line. AB - Heat-stable enterotoxin (STa) stimulates intestinal Cl(-) secretion by activating guanylate cyclase C (GCC) to increase intracellular cyclic GMP (cGMP). In the colon, cGMP action could involve protein kinase (PK) G-II or PKA pathways, depending on the segment and species. In the human colon, both PKG and PKA pathways have been implicated, and, therefore, the present study examined the mechanism of cGMP-mediated Cl(-) transport in primary cultures of human distal colonocytes and in T84, the colonic cell line. Both cell preparations express mRNA for CFTR, Na(+)-K(+)-2Cl(-) cotransporter (NKCC1), GCC and PKG-II as detected by RT-PCR. The effects of STa and the PKG-specific cGMP analogues, 8Br cGMP and 8pCPT-cGMP, on Cl(-) transport were measured using a halide-sensitive probe. In primary human colonocytes and T84 cells, STa, the cGMP analogues and the cAMP-dependent secretagogue, prostaglandin E(1) (PGE(1)), enhanced Cl(-) transport. The effects of 8Br-cGMP and 8pCPT-cGMP suggested the involvement of PKG, and this was explored further in T84 cells. The effects of 8pCPT-cGMP were dose-dependent and sensitive to the PKG inhibitor, H8 (70 microM), but H8 had no effect on PGE(1)-induced Cl(-) secretion. In contrast, a PKA inhibitor, H7 (50 microM), blocked PGE(1)-mediated but not 8pCPT-cGMP-induced Cl(-) transport. 8pCPT-cGMP enhanced phosphorylation of the PKG-specific substrate, 2A3, by T84 membranes in vitro. This phosphorylation was inhibited by H8. These results strongly suggest that cGMP activates Cl(-) transport through a PKG-II pathway in primary cells and in the T84 cell line of the human colon. PMID- 11042349 TI - CHO cell enlargement oscillates with a temperature-compensated period of 24 min. AB - The rate of increase in cell area of CHO cells when measured at intervals of 1 min using a light microscope equipped with a video measurement system, oscillated with a minimum period of about 24 min. The pattern of oscillations paralleled those of the 24 min period observed with the oxidation of NADH by an external cell surface or plasma membrane NADH oxidase. The increase in cell area was non linear. Intervals of rapid increase in area alternated with intervals of rapid decrease in area. The length of the 24 min period was temperature-compensated (approximately the same when measured at 14 degrees C, 24 degrees C or 34 degrees C) while the rate of cell enlargement increased with temperature over this same range of temperatures. PMID- 11042350 TI - Periodic NADH oxidase activity associated with an endoplasmic reticulum fraction from pig liver. Response to micromolar concentrations of retinol. AB - An endoplasmic reticulum fraction from pig liver enriched in transitional endoplasmic reticulum vesicles capable of forming 50-60 nm buds in the presence of ATP and retinol was assayed for retinol-responsive oxidation of NADH and cleavage of a dithiodipyridine (DTDP) protein disulfide-thiol interchange substrate. Maxima for the two activities alternated giving rise to a 24 min period. The NADH oxidase activity was inhibited by micromolar and submicromolar concentrations of retinol. Retinol at 0.1 mM stimulated the activity. The inhibition was confined to two activity maxima separated in time by about 5 min. In contrast, with the DTDP substrate, the activity was stimulated by retinol and the stimulations were in the part of the oscillatory pattern where retinol inhibition of NADH oxidation was observed. The findings support an earlier proposed mechanism whereby retinol exerted opposing effects on NADH oxidation and protein disulfide reductions. PMID- 11042351 TI - Subcellular localization of cyclic ADP-ribosyl cyclase and cyclic ADP-ribose hydrolase activities in porcine airway smooth muscle. AB - Recent studies have provided evidence for a role of cyclic ADP-ribose (cADPR) in the regulation of intracellular calcium in smooth muscles of the intestine, blood vessels and airways. We investigated the presence and subcellular localization of ADP-ribosyl cyclase, the enzyme that catalyzes the conversion of beta-NAD(+) to cADPR, and cADPR hydrolase, the enzyme that degrades cADPR to ADPR, in tracheal smooth muscle (TSM). Sucrose density fractionation of TSM crude membranes provided evidence that ADP-ribosyl cyclase and cADPR hydrolase activities were associated with a fraction enriched in 5'-nucleotidase activity, a plasma membrane marker enzyme, but not in a fraction enriched in either sarcoplasmic endoplasmic reticulum calcium ATPase or ryanodine receptor channels, both sarcoplasmic reticulum markers. The ADP-ribosyl cyclase and cADPR hydrolase activities comigrated at a molecular weight of approximately 40 kDa on SDS-PAGE. This comigration was confirmed by gel filtration chromatography. Investigation of kinetics yielded K(m) values of 30.4+/-1.5 and 695. 3+/-171.2 microM and V(max) values of 330.4+/-90 and 102.8+/-17.1 nmol/mg/h for ADP-ribosyl cyclase and cADPR hydrolase, respectively. These results suggest a possible role for cADPR as an endogenous modulator of [Ca(2+)](i) in porcine TSM cells. PMID- 11042352 TI - Nitric oxide induces oxidative stress and apoptosis in neuronal cells. AB - Within the central nervous system and under normal conditions, nitric oxide (NO) is an important physiological signaling molecule. When produced in large excess, NO also displays neurotoxicity. In our previous report, we have demonstrated that the exposure of neuronal cells to NO donors induced apoptotic cell death, while pretreatment with free radical scavengers L-ascorbic acid 2-[3, 4-dihydro-2,5,7,8 tetramethyl-2-(4,8, 12-trimethyltridecyl)-2H-1-benzopyran-6-yl-hydrogen phosphate] potassium salt (EPC-K1) or superoxide dismutase attenuated apoptosis effectively, suggesting that reactive oxygen species (ROS) may be involved in the cascade of events leading to apoptosis. In the present investigation, we directly studied the kinetic generation of ROS in NO-treated neuronal cells by flow cytometry using 2', 7'-dichloro-fluorescein diacetate and dihydrorhodamine 123 as redox-sensitive fluorescence probes. The results indicated that exposure of cerebellar granule cells to the NO donor S-nitroso-N-acetylpenicillamine (SNAP) induced oxidative stress, which was characterized by the accumulation of cytosolic and mitochondrial ROS, the increase in the extracellular hydrogen peroxide level, and the formation of lipid peroxidation products. SNAP treatment also induced apoptotic cell death as confirmed by the formation of cytosolic mono and oligonucleosomes. Pretreating cells with the novel antioxidant EPC-K1 effectively prevented oxidative stress induced by SNAP, and attenuated cells from apoptosis. PMID- 11042353 TI - Muscarinic receptor subtypes expression in rat and chick dorsal root ganglia. AB - In the present work we have analyzed by Northern blot, RT-PCR and in situ hybridization the expression of muscarinic receptor subtype mRNAs in rat and chick dorsal root ganglia. Northern blot analysis performed on rat total RNA revealed a strong signal for M(2) while a faint band was observed for M(3) and M(4) subtypes; no signal was evident for M(1) and M(5), while in chick total RNA no signal was detected for any of the analyzed subtypes (M(2), M(3), M(4)). On the other hand, RT-PCR revealed that all muscarinic subtype mRNAs were present both in rat and chick DRG, although the level of their expression may be different. In chick DRG, the presence of various muscarinic subtypes was confirmed by competition binding experiments. In situ hybridization in rat DRG showed that M(3) and M(4) transcripts, similarly to what has been previously described for M(2) mRNA, were preferentially localized in medium-small neurons. Large neurons were usually negative or faintly labelled. No hybridization signal was detected in rat DRG with probes for M(1) and M(5) muscarinic subtypes. The presence of various muscarinic receptors in DRG and their preferential expression in the medium-small sensory neurons suggest their possible involvement in the modulation of nociceptive stimuli transduction. PMID- 11042354 TI - Identification of differentially expressed genes in the visual structures of brain using high-density cDNA grids. AB - The hybridization patterns of 18,371 high-density-grid-arrayed non-redundant complementary DNA (cDNA) clones were examined using three different sources of cDNA probes. The first set of probes was synthesized from mRNA isolated from visual brain areas MT and V4 of Vervet monkey. The second set of probes was derived from cDNA libraries constructed from two micro dissected sets of layers of the monkey Lateral Geniculate Nucleus layers within the visual pathway, namely the magnocellular and parvocellular layers. The third set of cDNA probes was synthesized from the subtracted fractions of the cDNAs enriched for either the magnocellular or the parvocellular layers of the Lateral Geniculate Nucleus. Software, linked directly to the Genbank database, was developed to aid in the rapid identification of both expressed and differentially expressed genes. Our results indicate that both the cDNA probes synthesized from mRNA and cDNA libraries can identify similar fractions of expressed genes. However, the subtracted cDNA probes improve the efficiency of detection for those genes that are expressed at much lower abundance. Analyses of these results for the differential expression patterns of these genes were validated by semi quantitative PCR on the DNA derived from the whole tissue cDNA libraries. A list of some known genes that are statistically differentially expressed within the magnocellular layers of the LGN and area MT in the primate visual areas is derived. PMID- 11042355 TI - Neuropeptide amidation: cloning of a bifunctional alpha-amidating enzyme from Aplysia. AB - One of the most common mechanisms of posttranslational modifications to generate biologically active (neuro)peptides is the process of peptide alpha-amidation. The only enzyme known to catalyze this important modification is peptidylglycine alpha-amidating monooxygenase (PAM): a (bifunctional) zymogen, giving rise to a monooxygenase (PHM) and a lyase (PAL). The highly peptidergic central nervous system and endocrine system of the marine mollusk Aplysia has homologs of various mammalian peptide processing enzymes, including furin, Afurin2, prohormone convertase 1 (PC1), PC2, carboxypeptidase E (CPE) and CPD. Previously, it has been shown that the abdominal ganglion of Aplysia, which contains approximately 800 peptidergic bag cell neurons, contains the highest specific alpha-amidating activity. We have identified and cloned multiple overlapping central nervous system and bag cell cDNAs that encode a predicted 748-residue protein that is a member of the PAM family. The protein sequence contains the contiguous sequence of the catalytic domains of PHM and PAL, clearly demonstrating the existence of bifunctional Aplysia PAM, the first invertebrate PAM zymogen with an organization similar to that in vertebrates. None of the characterized clones encoded the so called exon A domain between the PHM and PAL domains. Furthermore, in a specific search by reverse transcription-polymerase chain reaction of RNA from multiple tissues we could only detect exon A-less transcripts. PAM expression was detected in the central nervous system, and in several endocrine and exocrine organs. Aplysia PAM is a candidate prohormone processing enzyme that plays an important role in the processing of Aplysia prohormones in the secretory pathway. PMID- 11042356 TI - Structure, biological activity of the upstream regulatory sequence, and conserved domains of a middle molecular mass neurofilament gene of Xenopus laevis. AB - During development, the molecular compositions of neurofilaments (NFs) undergo progressive modifications that correlate with successive stages of axonal outgrowth. Because NFs are the most abundant component of the axonal cytoskeleton, understanding how these modifications are regulated is essential for knowing how axons control their structural properties during growth. In vertebrates ranging from lamprey to mammal, orthologs of the middle molecular mass NF protein (NF-M) share similar patterns of expression during axonal outgrowth, which suggests that these NF-M genes may share conserved regulatory elements. These elements might be identified by comparing the sequences and activities of regulatory domains among the vertebrate NF-M genes. The frog, Xenopus laevis, is a good choice for such studies, because its early neural development can be observed readily and because transgenic embryos can be made easily. To begin such studies, we isolated genomic clones of Xenopus NF-M(2), tested the activity of its upstream regulatory sequence (URS) in transgenic embryos, and then compared sequences of regulatory regions among vertebrate NF-M genes to search for conserved elements. Studies with reporter genes in transgenic embryos found that the 1. 5 kb URS lacked the elements sufficient for neuron specific gene expression but identified conserved regions with basal regulatory activity. These studies further demonstrated that the NF-M 1.5 kb URS was highly susceptible to positional effects, a property that may be relevant to the highly variant, tissue-specific expression that is seen among members of the intermediate filament gene family. Non-coding regions of vertebrate NF-M genes contained several conserved elements. The region of highest conservation fell within the 3' untranslated region, a region that has been shown to regulate expression of another NF gene, NF-L. Transgenic Xenopus may thus prove useful for testing further the activity of conserved elements during axonal development and regeneration. PMID- 11042357 TI - Differential regulation of two period genes in the Xenopus eye. AB - The recent identification and analysis of mammalian homologues of the well characterized Drosophila circadian clock gene, Period (Per), has led to the idea that key features of vertebrate circadian rhythmicity are conserved at the molecular level. The Xenopus laevis retina contains a circadian clock mechanism that can be studied in vitro. To study the rhythmic expression of Per in the Xenopus retina, we used a degenerate RT-PCR strategy to obtain cDNA clones covering the entire 1427 amino acid coding region of a Xenopus homologue of Per2 and a partial cDNA sequence for a Xenopus homologue of Per1. Northern blot analysis shows that xPer1 and xPer2 transcripts are expressed most abundantly in the eye and the brain. However, rhythmic expression of xPer2 transcripts in the retina and retinal pigment epithelium (RPE) is light dependent and occurs only under 12 h light/12 h dark (LD) conditions, not in constant dark (DD). In contrast, xPer1 mRNA accumulation is rhythmic under both LD and DD conditions. Light dependent regulation of xPer2 mRNA and circadian regulation of xPer1 mRNA in the Xenopus retina differs from that in Drosophila and mammals. Light dependence of xPer2 mRNA levels and the offset phase relationship of the xPer2 rhythm to that for xPer1 suggests a role for xPer2 in circadian entrainment. PMID- 11042359 TI - Cloning, localisation and functional expression of a novel human, cerebellum specific, two pore domain potassium channel. AB - We have isolated, by degenerate PCR, a complementary DNA encoding a novel two pore domain potassium channel. This is the 7th functional member of the human tandem pore domain potassium channel family to be reported. It has an open reading frame of 1.125 kb and encodes a 374 amino acid protein which shows 62% identity to the human TASK-1 gene: identity to other human members of the family is 31-35% at the amino acid level. We believe this gene to be human TASK-3, the ortholog of the recently reported rat TASK-3 gene: amino acid identity between the two is 74%. 'Taqman' mRNA analysis demonstrated a very specific tissue distribution pattern, showing human TASK-3 mRNA to be localised largely in the cerebellum, in contrast rat TASK-3 was reported to be widely distributed. We have shown by radiation hybrid mapping that human TASK-3 can be assigned to chromosome 8q24.3. Human TASK-3 was demonstrated to endow Xenopus oocytes with a negative resting membrane potential through the presence of a large K(+) selective conductance. TASK-3 is inhibited by extracellular acidosis with a mid-point of inhibition around pH 6. 5, supporting the predictions from the sequence data that this is a third human TASK (TWIK-related acid sensitive K(+) channel) gene. PMID- 11042358 TI - Corticosteroids regulate 5-HT(1A) but not 5-HT(1B) receptor mRNA in rat hippocampus. AB - The role of mineralocorticoid and glucocorticoid receptors (MR and GR, respectively) in the regulation of serotonin receptors has not been clearly delineated. There is no consensus regarding the regulation of 5-HT(1A) receptors, and corticosteroid regulation of 5-HT(1B) mRNA has not been previously studied. We compared the effects of long-term (two week) adrenalectomy (no MR or GR activation) and several hormone replacement protocols designed to stimulate MR selectively (ALDO), MR and GR (HCT), and continuous MR with cyclical GR activation (SHAM adrenalectomy). 5-HT(1A) and 5-HT(1B) mRNAs were measured by in situ hybridization in hippocampus and raphe nuclei. None of the experimental manipulations altered 5-HT(1B) mRNA levels in the hippocampus or dorsal raphe, and also had no effect on 5-HT(1A) mRNA in dorsal or median raphe. However, 5 HT(1A) mRNA levels were regulated in a complex manner in the different subfields of hippocampus. We conclude that both MR and GR play an integrated role in regulating 5-HT(1A) mRNA levels in hippocampus while having no effect on 5-HT(1B) mRNA levels under these conditions. PMID- 11042360 TI - Co-localisation, heterophilic interactions and regulated expression of IgLON family proteins in the chick nervous system. AB - The chick glycoprotein GP55 has been shown to inhibit the growth and adhesion of DRG and forebrain neurons. GP55 consists of several members of the IgLON family, a group of glycoproteins including LAMP, OBCAM, CEPU-1 (chick)/neurotrimin (rat) and neurotractin (chick)/kilon (rat) thought to play a role in the guidance of growing axons. IgLONs belong to the Ig superfamily and have three C2 domains and a glycosyl phosphatidylinositol anchor which tethers them to the neuronal plasma membrane. We have now completed the deduced amino acid sequence for two isoforms of chicken OBCAM and used recombinant LAMP, OBCAM and CEPU-1 to raise antisera specific to these three IgLONs. LAMP and CEPU-1 are co-expressed on DRG and sympathetic neurons, while both overlapping and distinct expression patterns for LAMP, OBCAM and CEPU-1 are observed in retina. Analysis of IgLON mRNA expression reveals that alternatively spliced forms of LAMP and CEPU-1 are developmentally regulated. In an attempt to understand how the IgLONs function, we have begun to characterise their molecular interactions. LAMP and CEPU-1 have already been shown to interact homophilically. We now confirm that OBCAM will bind homophilically and also that LAMP, OBCAM and CEPU-1 will interact heterophilically with each other. We propose that IgLON activity will depend on the complement of IgLONs expressed by each neuron. PMID- 11042361 TI - Schizophrenia: elevated mRNA for calcium-calmodulin-dependent protein kinase IIbeta in frontal cortex. AB - Because amphetamine releases two to three times more dopamine in schizophrenia patients than in control subjects, and because calcium-calmodulin-dependent protein kinase II has a key role in the enhanced action of amphetamine-induced dopamine release in rats, the synaptic content of calcium-calmodulin-dependent protein kinase IIbeta mRNA was measured (by quantitative competitive RT-PCR; reverse transcriptase-polymerase chain reaction) in seven frontal cerebral cortices of post-mortem brains from patients who had schizophrenia and in seven control tissues. The results indicate that the mRNA of this kinase is elevated in the schizophrenia frontal cortex. PMID- 11042362 TI - Region-specific decline in the expression of metabotropic glutamate receptor 7 mRNA in rat brain during aging. AB - Age-dependent changes in the expression of group III metabotropic glutamate receptors (mGluR4 and mGluR7) were studied by quantitative in situ hybridization using male Fisher 344 rats 3, 12 and 25 months of age. Results indicate an early decrease in mGluR7 mRNA level in several cortical areas including the frontal, parietal and temporal cortices. In the hippocampus, mGluR7 mRNA levels decreased in the CA1 region and the lower blade of the dentate gyrus. Moreover, significant decrease was found in the laterodorsal thalamic nucleus at 12 months of age. Other regions such as the caudate putamen and nucleus accumbens showed no age related changes in mGluR7 mRNA levels. Analysis of emulsion autoradiograms revealed a 36% decrease of mGluR7 mRNA in Purkinje neurons in the 12-month-old group and a 48% decline in the 25-month-old group as compared to the 3-month-old group. A substantial decrease in mGluR4 mRNA level was found in the granule cell layer of the cerebellum during aging. The difference between the young and aged groups exceeded 35%. These region-specific decreases may have important implication in some of the age-related changes in cognitive, motor and/or sensory functions. PMID- 11042363 TI - Protein kinase C iota protects neural cells against apoptosis induced by amyloid beta-peptide. AB - Protein kinase C (PKC) isoforms are increasingly recognized as playing important roles in the regulation of neuronal plasticity and survival. Recent findings from studies of non-neuronal cells suggest that atypical isoforms of PKC can modulate apoptosis in various paradigms. Because increasing data support a role for neuronal apoptosis in the pathogenesis of Alzheimer's disease (AD), we tested the hypothesis that PKCiota (PKCiota) can modify vulnerability of neural cells to apoptosis induced by amyloid beta-peptide (ABP), a cytotoxic peptide linked to neuronal degeneration in AD. Overexpression of PKCiota increased the resistance of PC12 cells to apoptosis induced by ABP. Associated with the increased resistance to apoptosis were improved mitochondrial function and reduced activity of caspases. In addition, ABP-induced increases in levels of oxidative stress and intracellular calcium levels were attenuated in cells overexpressing PKCiota. These findings suggest that PKCiota prevents apoptosis induced by ABP by interrupting the cell death process at a very early step, thereby allowing the cells to maintain ion homeostasis and mitochondrial function. PMID- 11042364 TI - First localisation of somatostatin sst(4) receptor protein in selected human brain areas: an immunohistochemical study. AB - Somatostatin is known to have diverse neurophysiological effects in the mammalian CNS. To date, genes for five different receptors, termed sst(1-5), have been isolated. Recently several reports have been published on the localisation of the individual receptor protein in the rat CNS, but their localisation in the human CNS remains largely unknown. Until now little information about the function of the sst(4) receptor is available, and there is a lack of receptor specific agonists and antagonists. Here, we report for the first time the immunohistochemical localisation of the sst(4) receptor in selected human brain areas using an anti-peptide antibody raised against a carboxy-terminal portion of the receptor protein. Strong receptor immunoreactivity was found in several brain regions, including the hippocampal formation, the cerebellar cortex and the medulla. Further immunohistochemical labelling was observed in the cerebral cortex, the red nucleus and the globus pallidus. Somatodendritic as well as axonal staining was observed. Specific signals were entirely absent following antibody pre-adsorption with the synthetic peptide. The results are in good agreement with the previously published immunohistochemical localisation of the sst(4) receptor in the rat brain. This is the first immunohistochemical study of the localisation of the sst(4) receptor in the human brain, and implicates this receptor in the function of higher centres of the human nervous system. PMID- 11042365 TI - Thyroid hormone of maternal origin regulates the expression of RC3/neurogranin mRNA in the fetal rat brain. AB - Recent clinical studies indicate that thyroid hormone plays essential roles in fetal brain development. However, the mechanism by which thyroid hormone affects fetal brain development is poorly studied. We recently identified several genes expressed in the fetal cortex whose abundance is affected by thyroid hormone of maternal origin. However, it is unclear whether these genes are directly regulated by thyroid hormone. Because these are the first genes known to be regulated by thyroid hormone during fetal development, we sought to expand our investigation to genes known to be regulated directly by thyroid hormone. We now report that the well-known thyroid hormone-responsive gene RC3/neurogranin is expressed in the fetal brain and is regulated by thyroid hormone of maternal origin. These findings support the concept that maternal thyroid hormone exerts a direct action on the expression of genes in the fetal brain that are important for normal neurological development. PMID- 11042366 TI - TH protein and mRNA in nigrostriatal dopaminergic neurons are down-regulated by continuous but not intermittent apomorphine. AB - Levels of tyrosine hydroxylase (TH) and TH mRNA were measured after administration of dopamine agonists for a long period of time to elucidate the long-term feedback inhibition of dopamine synthesis in nigrostriatal dopaminergic neurons. Continuous infusion, which desensitized presynaptic dopamine receptors, but not repeated administration, down-regulated TH and TH mRNA levels. This suggests levels of TH protein and mRNA are only feedback inhibited by the continuous stimulation of postsynaptic dopamine receptors. PMID- 11042367 TI - Longitudinal gradients of KCNQ4 expression in spiral ganglion and cochlear hair cells correlate with progressive hearing loss in DFNA2. AB - Mutations in the human KCNQ4 gene were recently found by Kubisch et al. [Cell 96 (1999) 437-446] to cause a non-syndromic, autosomal dominant, progressive hearing loss, DFNA2. The mouse Kcnq4 orthologue was previously localized to the outer hair cells (OHCs) of the inner ear, suggesting the pathophysiological effects were due to dysfunctional OHCs. Yet, OHC dysfunction does not provide a plausible explanation for the progressive nature of the frequency specific hearing loss. We have re-examined and extended the expression analyses of KCNQ4 in the murine inner ear using RT-PCR and whole mount in situ hybridization. Our results confirmed that the rat KCNQ4 orthologue is expressed in both inner and outer hair cells. Reciprocal longitudinal gradients were found in inner hair cells (IHCs) and OHCs. The strongest expression of KCNQ4 in IHCc was in the base of the cochlea and in the apex for OHCs. Similar to the IHCs, a basal to apical gradient was present in the spiral sensory neurons. IHCs mediate hearing via their afferent sensory neurons, whereas OHCs function as active cochlear amplifiers. The complete absence of OHCs leads only to severe sensitivity reduction, but not complete hearing loss. Our data suggest that the primary defect leading to initial high frequency loss and subsequent progressive hearing loss for all frequencies may be due to spiral ganglion and/or IHC dysfunction, rather than an OHC aberration. PMID- 11042368 TI - Photoreceptor regulated expression of Ca(2+)/calmodulin-dependent protein kinase II in the mouse retina. AB - The objective of this investigation is to determine mechanisms for regulation of retinal calmodulin kinase II (CaMKII). To this end, the expression and activity of CaMKII are examined in the retina of the rdta mouse, in which rod photoreceptors have been genetically ablated [47]. CaMKII levels are compared between rdta mice and the normal, littermate control mice. It is demonstrated that retinal CaMKII protein, enzyme activity and mRNA are significantly increased in response to the genetic ablation of rod photoreceptors. The data indicate that CaMKII expression/activity in amacrine and ganglion cells is negatively regulated by the rod photoreceptor-mediated visual input. The regulation appears to occur primarily at the transcriptional level. It is shown that the cytoplasmic polyadenylation element binding protein (CPEB), a regulatory factor for translation that is known to promote CaMKIIalpha translation in dendrites [83], is also present in the mouse retina. However, the polyadenylation-mediated translational control mechanism is not activated in this experimental paradigm. PMID- 11042376 TI - Inactivation of a novel three-cistronic operon tcaR-tcaA-tcaB increases teicoplanin resistance in Staphylococcus aureus. AB - A novel teicoplanin-associated operon termed tcaR-tcaA-tcaB was identified by Tn917-mediated insertional mutagenesis. Resistance to teicoplanin rose 4-fold by insertional inactivation of tcaA or by deletion of the entire operon. tcaA encodes a hypothetical transmembrane protein with a metal-binding motif, possibly a sensor-transducer. tcaB codes for a membrane-associated protein, which has sequence homologies to a bicyclomycin resistance protein. The two genes are preceded by tcaR encoding a putative regulator with sequence homologies to the transcriptional regulator MarR. The fact that tcaA inactivation as well as deletion of tcaRAB produced the same increase in teicoplanin resistance confirmed the association of tcaRAB with teicoplanin susceptibility. Cotransductional crosses showed that the level of teicoplanin resistance produced by these insertions was strain-dependent and that in the methicillin-resistant strain COL, it was paired with a remarkable decrease in methicillin resistance. This allowed to postulate that tcaRAB may be involved in some way in cell wall biosynthesis, and that teicoplanin may interact with TcaA and/or TcaB either directly or indirectly. PMID- 11042377 TI - Melatonin reduces rat hepatic macromolecular damage due to oxidative stress caused by delta-aminolevulinic acid. AB - Delta-aminolevulinic acid, precursor of heme, accumulates in a number of organs, especially in the liver, of patients with acute intermittent porphyria. The potential protective effect of melatonin against oxidative damage to nuclear DNA and microsomal and mitochondrial membranes in rat liver, caused by delta aminolevulinic acid, was examined. Changes in 8-hydroxy-2'-deoxyguanosine (8 OHdG) levels, an index of DNA damage, and alterations in membrane fluidity (the inverse of membrane rigidity) and lipid peroxidation in microsomal and mitochondrial membranes, as indices of damage to lipid and protein molecules in membranes, were estimated. Measurements were made in rat liver after a 2 week treatment with delta-aminolevulinic acid (40 mg/kg b.w., every other day). To test the potential protective effects of melatonin, the indole was injected (i.p. 10 mg/kg b.w.) 3 times daily for 2 weeks. 8-OHdG levels and lipid peroxidation in microsomal membranes increased significantly whereas microsomal and mitochondrial membrane fluidity decreased as a consequence of delta-aminolevulinic acid treatment. Melatonin completely counteracted the effects of delta-aminolevulinic acid. Melatonin was highly effective in protecting against oxidative damage to DNA as well as to microsomal and mitochondrial membranes in rat liver and it may be useful as a cotreatment in patients with acute intermittent porphyria. PMID- 11042378 TI - On the modulation of photoinduced fluorescence enhancement and conformational stability of albumin-bound bilirubin: effect of epsilon-NH(2) groups blocking and chloroform binding. AB - Photoinduced fluorescence enhancement of bilirubin bound to primary binding site on human serum albumin (HSA) was completely ceased when epsilon-NH(2) groups of its internal lysine residues were covalently blocked by acetylation or succinylation though the pigment bound to these derivatives in a folded conformation akin to that bound to HSA. These photoinduced fluorescence modulations cannot be ascribed to the binding of bilirubin to secondary low affinity sites as the CD spectrum of bilirubin bound to these derivatives showed complete inversion upon addition of chloroform which binds to subdomain IIA in HSA where high affinity bilirubin binding site is located. Presence of chloroform reconciled the photoinduced alterations in the CD spectrum observed in its absence, suggesting that chloroform stabilized the bound ligand against light but the fluorescence properties of bilirubin complexed with acetylated or succinylated derivatives remained unchanged. Guanidination of internal epsilon NH(2) groups in HSA by O-methylisourea did not alter the spectral properties of the bound ligand. These results suggest that salt linkage(s) existing between epsilon-NH(2) groups of lysine residues in HSA and carboxyl groups of bilirubin, act(s) as a potential barrier during conformational rotation of the bound ligand assisted by photoactivation and their abolishment can alter its dynamics and stereoselectivity, a hitherto unnoticed implication of salt linkage(s) in BR-HSA complex. PMID- 11042379 TI - The iron chelator pyridoxal isonicotinoyl hydrazone (PIH) and its analogues prevent damage to 2-deoxyribose mediated by ferric iron plus ascorbate. AB - Iron chelating agents are essential for treating iron overload in diseases such as beta-thalassemia and are potentially useful for therapy in non-iron overload conditions, including free radical mediated tissue injury. Deferoxamine (DFO), the only drug available for iron chelation therapy, has a number of disadvantages (e.g., lack of intestinal absorption and high cost). The tridentate chelator pyridoxal isonicotinoyl hydrazone (PIH) has high iron chelation efficacy in vitro and in vivo with high selectivity and affinity for iron. It is relatively non toxic, economical to synthesize and orally effective. We previously demonstrated that submillimolar levels of PIH and some of its analogues inhibit lipid peroxidation, ascorbate oxidation, 2-deoxyribose degradation, plasmid DNA strand breaks and 5,5-dimethylpyrroline-N-oxide (DMPO) hydroxylation mediated by either Fe(II) plus H(2)O(2) or Fe(III)-EDTA plus ascorbate. To further characterize the mechanism of PIH action, we studied the effects of PIH and some of its analogues on the degradation of 2-deoxyribose induced by Fe(III)-EDTA plus ascorbate. Compared with hydroxyl radical scavengers (DMSO, salicylate and mannitol), PIH was about two orders of magnitude more active in protecting 2-deoxyribose from degradation, which was comparable with some of its analogues and DFO. Competition experiments using two different concentrations of 2-deoxyribose (15 vs. 1.5 mM) revealed that hydroxyl radical scavengers (at 20 or 60 mM) were significantly less effective in preventing degradation of 2-deoxyribose at 15 mM than 2 deoxyribose at 1.5 mM. In contrast, 400 microM PIH was equally effective in preventing degradation of both 15 mM and 1.5 mM 2-deoxyribose. At a fixed Fe(III) concentration, increasing the concentration of ligands (either EDTA or NTA) caused a significant reduction in the protective effect of PIH towards 2 deoxyribose degradation. We also observed that PIH and DFO prevent 2-deoxyribose degradation induced by hypoxanthine, xanthine oxidase and Fe(III)-EDTA. The efficacy of PIH or DFO was inversely related to the EDTA concentration. Taken together, these results indicate that PIH (and its analogues) works by a mechanism different than the hydroxyl radical scavengers. It is likely that PIH removes Fe(III) from the chelates (either Fe(III)-EDTA or Fe(III)-NTA) and forms a Fe(III)-PIH(2) complex that does not catalyze oxyradical formation. PMID- 11042380 TI - Introduction of the human growth hormone gene into the guinea pig mammary gland by in vivo transfection promotes sustained expression of human growth hormone in the milk throughout lactation. AB - We tested the feasibility of transfecting mammary tissue in vivo with an expression plasmid encoding the human growth hormone (hGH) gene, under the control of the cytomegalovirus promoter. Guinea pig mammary glands were transfected with plasmid DNA infused through the nipple canal and expression was monitored in control and transfected glands by radioimmunoassay of milk samples for hGH. Sustained expression of hGH throughout lactation was attained with a polyion transfection complex shown to be optimal for the transfection of bovine mammary cells, in vitro. However, contrary to expectations, hGH expression was consistently 5- to 10-fold higher when DEAE-dextran was used alone for transfection. Thus polyion complexes which are optimal for the transfection of cells in vitro may not be optimal in vivo. The highest concentrations of hGH in milk were obtained when glands were transfected within 3 days before parturition. This method may have application for studying the biological role or physical properties of recombinant proteins expressed in low quantities, or for investigating the regulation of gene promoters without the need to construct viral vectors or produce transgenic animals. PMID- 11042381 TI - The biochemical mode of inhibition of DNA polymerase beta by alpha-rubromycin. AB - Quinone antibiotics, alpha- and beta-rubromycin, were originally found as inhibitors of retroviral reverse transcriptase. We investigated the effects of these agents on DNA metabolic enzymes including DNA and RNA polymerases as retroviral reverse transcriptase is a kind of the polymerase. As expected, we found that alpha- and beta-rubromycin strongly inhibited not only the retroviral reverse transcriptase activity, but the activities of the mammalian DNA polymerases, telomerase and terminal deoxynucleotidyl transferase in vitro. These agents should therefore be classified as DNA polymerase inhibitors. The Ki values of alpha-rubromycin against nucleotide substrate were 0.66 and 0.17 microM for DNA polymerase alpha and beta (pol. alpha and beta), respectively, and those of beta-rubromycin was 2.40 and 10.5 microM, respectively. Alpha-rubromycin strongly inhibited the pol. beta activity, and showed the strongest pol. beta inhibitory effect reported to date. At least on pol. beta, alpha-rubromycin was suggested to bind to the active region competing with the nucleotide substrate, and subsequently inhibit the catalytic activity. alpha-Rubromycin directly competed with the nucleotide substrate, and indirectly but simultaneously and non competitively disturbed the template-DNA interaction with pol. beta. PMID- 11042382 TI - Differed preferential iron-binding lobe in human transferrin depending on the presence of bicarbonate detected by HPLC/high-resolution inductively coupled plasma mass spectrometry. AB - The binding of iron (Fe) to human serum transferrin (Tf) was analyzed with an HPLC system equipped with an anion exchange column and directly connected with a high-resolution inductively coupled plasma mass spectrometer for metal detection. The (56)Fe level in the eluate was monitored at resolution m/Deltam=3000. Two monoferric Tfs were assigned based on the results of urea-PAGE and desferrioxamine experiments. When Fe was added as Fe-citrate stepwise to an apo Tf solution in the presence of bicarbonate, the N-lobe site was the preferential Fe-binding site, while the C-lobe site was preferred in the absence of bicarbonate. In both cases, the Fe-peak areas of the preferential site and Fe(2) Tf increased up to an Fe/Tf molar ratio of 1, and then the peak area of the monoferric Tf decreased while the peak area of Fe(2)-Tf increased. When the Fe/Tf molar ratio was below 1, the amount of Fe bound to the lobe with a weaker affinity was higher in Fe(2)-Tf than in the monoferric Tf in each case. Namely, Fe(2)-Tf was the preferential binding state of Fe to human serum Tf. The preference is reasonable for transferring Fe ions effectively to Tf-receptors. PMID- 11042383 TI - Dopamine-melanin protects against tyrosine nitration, tryptophan oxidation and Ca(2+)-ATPase inactivation induced by peroxynitrite. AB - The effects of dopamine-melanin (DA-melanin), a synthetic model of neuromelanin, on peroxynitrite-mediated 3-nitrotyrosine formation, oxidation of tryptophan in bovine serum albumin and inactivation of erythrocyte membrane Ca(2+)-ATPase activity were investigated in the absence and in the presence of bicarbonate. DA melanin inhibited nitration of free tyrosine, loss of tryptophan residues and Ca(2+)-ATPase inactivation by peroxynitrite in a dose dependent manner. In the presence of bicarbonate, this inhibitory effect was lower for nitration and insignificant for oxidative protein modifications. These results suggest that neuromelanin can protect against nitrating and oxidizing action of peroxynitrite but is a worse protector against the peroxynitrite-CO(2) adduct. As peroxynitrite may be a mediator of neurotoxic processes, the obtained results suggest that neuromelanin may be important as a physiological protector against peroxynitrite. PMID- 11042384 TI - Ovotransferrin antimicrobial peptide (OTAP-92) kills bacteria through a membrane damage mechanism. AB - Ovotransferrin antimicrobial peptide (OTAP-92) is a cationic fragment of hen ovotransferrin (OTf). OTAP-92 consists of 92 amino acid residues located within the 109-200 sequence of the N-lobe of OTf. This study was aimed to delineate the antimicrobial mechanism of OTAP-92 and to identify its interaction with bacterial membranes. OTAP-92 caused permeation of Escherichia coli outer membrane (OM) to 1 N-phenylnaphthylamine fluorescent probe in a dose-dependent manner. These results suggested that OTAP-92 crossed the bacterial OM by a self-promoted uptake. Cytoplasmic membrane of E. coli was found to be the target for OTAP-92 bactericidal activity, as assayed by the unmasking of cytoplasmic beta galactosidase due to membrane permeabilization in a kinetic manner. Pretreatment of bacteria with uncoupler, carbonyl cyanide m-chlorophenylhydrazone, markedly enhanced permeation of cytoplasmic membrane, suggesting that the membrane permeation due to OTAP-92 is independent of the transmembrane potential. In an E. coli phospholipid liposome model, it was demonstrated that OTAP-92 has the ability to dissipate the transmembrane electrochemical potential. Intrinsic fluorescence spectra of the two tryptophan residues in OTAP-92, using liposomal membrane, have identified the lipid-binding region as a helix-sheet motif, and suggested an adjacent Ca(2+)-sensitive site within OTAP-92. These data indicated that OTAP-92 possesses a unique structural motif similar to the insect defensins. Further, this cationic antimicrobial peptide is capable of killing Gram-negative bacteria by crossing the OM by a self-promoted uptake and cause damage to the biological function of cytoplasmic membrane. PMID- 11042385 TI - Determination of H(2)S solubility via the reaction with ferric hemoglobin I from the bivalve mollusc Lucina pectinata. AB - A new, simple and fast spectrophotometric method for the determination of the H(2)S concentration is reported. This method, based on the 1:1 reaction between H(2)S and the ferric derivative of hemoglobin I (HbI) from the bivalve mollusc Lucina pectinata, allows the quantitative determination of H(2)S dissolved in a given solution even at concentrations as low as 1 x 10(-6) M. Note that L. pectinata HbI is considered the physiological receptor of H(2)S. PMID- 11042386 TI - Time course and quantitative analysis of the adaptive responses to 85% oxygen in the rat lung and heart. AB - The phenomenon of oxygen tolerance (resistance to 100% O(2) in rats previously exposed to 85% O(2)) constitutes one of the few models of adaptive responses to oxidative stress in mammals. In vitro studies suggest that reactive oxygen species mediate this response. To test this hypothesis in vivo, we followed the time course of oxidative stress, enzyme induction, and edema in the lung, heart and liver of rats exposed to 85% O(2) for 1 to 5 days. Interestingly, not only the lung, but also the heart of rats exposed to 85% O(2) showed increases in the production of O(*-)(2) (aconitase inactivation) early during the exposure. Increases in O(*-)(2) were associated to oxidative stress (increased in situ chemiluminescence) and transient edema in both tissues. Both the lung and heart displayed induction of superoxide dismutase and reversion of the oxidative stress and damage. The adaptive response in the heart was faster and more efficient, suggesting that this tissue is at a more critical risk when exposed to elevated O(2) concentrations. PMID- 11042387 TI - Modulation of luteinizing hormone subunit gene expression by intracerebroventricular microinjection of gonadotropin-releasing hormone or beta endorphin in female rats. AB - The effects of gonadotropin-releasing hormone (GnRH), beta-endorphin and its antagonist naloxone on the expression of luteinizing hormone (LH) subunit genes and LH secretion were examined in ovariectomized and/or cycling female rats through their direct microinjection into the third cerebral ventricle, in the proximity of the hypothalamus-pituitary complex. GnRH (1 nM) induced a significant augmentation of the pituitary content of alpha mRNA when administered 15, 30 or 60 min intervals over 5 h to ovariectomized rats whereas only the 30 and 60 min intervals were effective in increasing LHbeta mRNA, and the 60 min intervals for LH release. This was in agreement with the established concept of a pulse-dependent regulation of gonadotropin synthesis and release. Hourly pulses of GnRH also increased alpha and LHbeta mRNA levels when microinjected in female cycling rats during proestrus or diestrus II. Using this model we observed a marked negative influence of hourly intracerebral microinjections of beta endorphin on LH mRNA content and LH release in ovariectomized rats while naloxone had no effect. This suggests that endogenous beta-endorphin was unable to exert its negative action on beta-endorphin receptors that were present and responded to the ligand. The present approach would be valuable for the exploration of the mechanisms of action of beta-endorphin or other substances on the functions of the gonadotrophs. PMID- 11042388 TI - Hemolysis of human red blood cells by riboflavin-Cu(II) system. AB - The photodynamic action of riboflavin is generally considered to involve the generation of reactive oxygen species, whose production is enhanced when Cu(II) is present in the reaction. In the present study we report that photoactivated riboflavin causes K(+) loss from fresh human red blood cells (RBC) in a time dependent manner. Addition of Cu(II) further enhances the K(+) loss and also leads to significant hemolysis. Riboflavin in a 2:1 stoichiometry with Cu(II) leads to maximum K(+) loss and up to 45% hemolysis. Bathocuproine, a specific Cu(I)-sequestering agent, when present in the reaction, inhibits the hemolysis completely. Free radical scavengers like superoxide dismutase, potassium iodide and mannitol inhibited the hemolysis up to 55% or more. However, thiourea was the most effective scavenger showing 90% inhibition. These results suggest that K(+) leakage and hemolysis of human RBC are basically free radical mediated reactions. PMID- 11042389 TI - Specific ganglioside changes in extraneural tissues of adult rats with hypothyroidism. AB - Adults rats with hypothyroidism were prepared by administration of 6-propyl-2 thiouracil (PTU) or methimazole, and the tissues were examined for their gangliosides through methods including glycolipid-overlay techniques. Normal thyroid tissue contained GM3, GD3, and GD1a as the major gangliosides, with GM1, GD1b, GT1b, and GQ1b in lesser amounts. The goitrous tissue of PTU-induced hypothyroid rats had higher concentrations of GM1 and GD1a with a concomitant decrease of GM3. The amount of GT3 in thyroid tissue was increased in hypothyroid animals. While normal liver tissue had a complex ganglioside pattern with a- and b-series gangliosides, the PTU-induced hypothyroid tissue showed a simpler ganglioside profile that consisted mainly of a-series gangliosides with almost undetectable amounts of b-series gangliosides. The expression of c-series gangliosides was suppressed in the hypothyroid liver tissue. Heart tissue had higher contents of GM3 and GT3 than control. No apparent change was observed in the compositions of major and c-series gangliosides in other extraneural tissues (i.e., kidney, lung, spleen, thymus, pancreas, testis, skeletal muscle, and eye lenses), and neural tissues (i.e., cerebrum and cerebellum) from PTU-induced hypothyroid rats. The ganglioside changes of thyroid, liver, and heart tissues were reproduced in corresponding tissues of methimazole-induced hypothyroid rats. These results suggest that hypothyroid conditions affect the biosynthesis and expression of gangliosides in specific tissue and cell types. PMID- 11042390 TI - Structural analysis of N-glycans from yellow lupin (Lupinus luteus) seed diphosphonucleotide phosphatase/phosphodiesterase. AB - N-linked oligosaccharide chains released by hydrazinolysis from yellow lupin seed diphosphonucleotide phosphatase/phosphodiesterase were fluorescence labeled and separated by high performance liquid chromatography (GlycoSep N and GlycoSep H columns). Exoglycosidase sequencing elucidated the structures of 24 separated N glycans. Thirty percent of isolated glycans were found to be of high-mannose type (three to eight mannosyl residues), 42% were complex type and 26% belonged to paucimannosidic type. Among complex type glycans, structures with Lewis(a) epitope were identified. It is very unusual to find all types of plant N-glycans in one protein. Possible reasons for such a broad spectrum of N-glycan structures are discussed. PMID- 11042392 TI - Amino acid sequence and some properties of phytolacain G, a cysteine protease from growing fruit of pokeweed, Phytolacca americana. AB - A protease, phytolacain G, has been found to appear on CM-Sepharose ion-exchange chromatography of greenish small-size fruits of pokeweed, Phytolacca americana L, from ca. 2 weeks after flowering, and increases during fruit enlargement. Reddish ripe fruit of the pokeweed contained both phytolacain G and R. The molecular mass of phytolacain G was estimated to be 25.5 kDa by SDS-PAGE. Its amino acid sequence was reconstructed by automated sequence analysis of the peptides obtained after cleavage with Achromobacter protease I, chymotrypsin, and cyanogen bromide. The enzyme is composed of 216 amino acid residues, of which it shares 152 identical amino acid residues (70%) with phytolacain R, 126 (58%) with melain G, 108 (50%) with papain, 106 (49%) with actinidain, and 96 (44%) with stem bromelain. The amino acid residues forming the substrate binding S(2) pocket of papain, Tyr67, Pro68, Trp69, Val133, and Phe207, were predicted to be replaced by Trp, Met, His, Ala, and Ser in phytolacain G, respectively. As a consequence of these substitutions, the S(2) pocket is expected to be less hydrophobic in phytolacain G than in papain. PMID- 11042391 TI - Chorion peroxidase-mediated NADH/O(2) oxidoreduction cooperated by chorion malate dehydrogenase-catalyzed NADH production: a feasible pathway leading to H(2)O(2) formation during chorion hardening in Aedes aegypti mosquitoes. AB - A specific chorion peroxidase is present in Aedes aegypti and this enzyme is responsible for catalyzing chorion protein cross-linking through dityrosine formation during chorion hardening. Peroxidase-mediated dityrosine cross-linking requires H(2)O(2), and this study discusses the possible involvement of the chorion peroxidase in H(2)O(2) formation by mediating NADH/O(2) oxidoreduction during chorion hardening in A. aegypti eggs. Our data show that mosquito chorion peroxidase is able to catalyze pH-dependent NADH oxidation, which is enhanced in the presence of Mn(2+). Molecular oxygen is the electron acceptor during peroxidase-catalyzed NADH oxidation, and reduction of O(2) leads to the production of H(2)O(2), demonstrated by the formation of dityrosine in a NADH/peroxidase reaction mixture following addition of tyrosine. An oxidoreductase capable of catalyzing malate/NAD(+) oxidoreduction is also present in the egg chorion of A. aegypti. The cooperative roles of chorion malate/NAD(+)oxidoreductase and chorion peroxidase on generating H(2)O(2) with NAD(+) and malate as initial substrates were demonstrated by the production of dityrosine after addition of tyrosine to a reaction mixture containing NAD(+) and malate in the presence of both malate dehydrogenase fractions and purified chorion peroxidase. Data suggest that chorion peroxidase-mediated NADH/O(2) oxidoreduction may contribute to the formation of the H(2)O(2) required for chorion protein cross-linking mediated by the same peroxidase, and that the chorion associated malate dehydrogenase may be responsible for the supply of NADH for the H(2)O(2) production. PMID- 11042393 TI - Comparative characterization of two serine endopeptidases from Nocardiopsis sp. NCIM 5124. AB - A protease-producing, crude oil degrading marine isolate was identified as Nocardiopsis sp. on the basis of the morphology, cell wall composition, mycolic acid analysis and DNA base composition. The Nocardiopsis produces two extracellular proteases, both of which are alkaline serine endopeptidases. Protease I was purified to homogeneity by chromatography on CM-Sephadex at pH 5.0 and pH 9.0. Protease II was purified using DEAE-cellulose, Sephadex G-50, phenyl Sepharose and hydroxyapatite chromatography. Protease I and II had almost similar M(r) of 21 kDa (Protease I) and 23 kDa (Protease II), pI of 8.3 and 7.0 respectively with pH and temperature optima for activity between 10.0 and 11.0 and about 60 degrees C. Specific activities were 152 and 14 U/mg respectively on casein. However, Protease I was antigenically unrelated to Protease II. Both proteases were endopeptidases and required extended substrate binding for catalysis. Both proteases had collagenolytic and fibrinolytic activity but only Protease I had elastinolytic activity. The proteases were chymotrypsin-like with respect to their amino acid compositions and N-terminal sequences. PMID- 11042394 TI - Functional expression and genomic structure of human N-acetylglucosamine-6-O sulfotransferase that transfers sulfate to beta-N-acetylglucosamine at the nonreducing end of an N-acetyllactosamine sequence. AB - The cDNA and gene encoding human N-acetylglucosamine-6-O-sulfotransferase (Gn6ST) have been cloned. Comparative analysis of this cDNA with the mouse Gn6ST sequence indicates 96% amino acid identity between the two sequences. The expression of a soluble recombinant form of the protein in COS-1 cells produced an active sulfotransferase, which transferred sulfate to the terminal GlcNAc in GlcNAcbeta1 O-CH(3), GlcNAcbeta1-3Galbeta1-O-CH(3) and GlcNAcbeta1-3Galbeta1-4GlcNAcbeta1 3Galbeta1-4Gl cNAc but not in GlcNAcalpha1-4GlcAbeta1-3Galbeta1-3Galbeta1-4 Xylbeta1-O-Ser. In addition, neither Galbeta1-4GlcNAcbeta1-O-naphthalenemethanol nor GalNAcbeta1-4GlcAbeta1-3Galbeta1-3Galbeta1-4X ylbeta1-O-Ser were utilized as acceptors. These findings indicate that a terminal beta-linked GlcNAc residue is necessary for acceptor substrates of Gn6ST. The human Gn6ST gene spans about 7 kb, consists of two exons and exhibits an intron-less coding region. PMID- 11042395 TI - Proceedings of the European Society for Veterinary Virology 4th Pestivirus Meeting. Giessen, Germany, 15-19 March 1999. PMID- 11042396 TI - The 1997/1998 epizootic of swine fever in the Netherlands: control strategies under a non-vaccination regimen. AB - The 1997/1998 epizootic of classical swine fever (CSF) in an area with high pig density in the Netherlands is described. The epizootic, which numbered 429 outbreaks, was controlled and finally eradicated after 14 months without resorting to vaccination. A further almost 1300 herds (1.1 million pigs) at close proximity of confirmed outbreaks were preventively culled because of the risk of having been infected. The pros and cons of this so-called "pre-emptive slaughter" are discussed. The long-lasting movement restrictions caused severe overcrowding especially in breeding farms. For reasons of animal welfare 6.5 million weaners and adult pigs had to be killed and destroyed, whereas another 2.6 million 3-17 days old piglets were euthanised to save long-term destruction capacity. The presumed routes of infection and factors influencing the epizootic are explained, as well as the various methods to bring the epizootic to a halt. The strategy for detecting outbreaks in an early stage, and the type of samples to be collected for laboratory diagnosis are emphasised from the general point of application. The direct costs of the epizootic, losses of exports not included, are estimated at US$ 2 billion. PMID- 11042397 TI - Molecular epidemiology of a large classical swine fever epidemic in the European Union in 1997-1998. AB - A big epidemic of classical swine fever (CSF) occurred in the European Community in 1997. The first case was reported at the beginning of January 1997 from Germany. The disease presumably spread to the Netherlands, and from there to Italy, Spain and eventually to Belgium. About 30 isolates from these outbreaks were analysed by comparison of the nucleotide sequence data generated from fragments of both the E2 glycoprotein gene (190 nucleotides) and from the 5' nontranslated region (5'-NTR; 150 nucleotides). By combining epidemiological data with genetic typing, it was found that the outbreaks were related and caused by a virus belonging to the genetic subgroup 2.1. As this type of virus had been reported infrequently in Europe and not at all since 1993, we postulate that it was newly introduced into the European Union (EU). PMID- 11042398 TI - Epidemiology of classical swine fever in Germany in the 1990s. AB - In Germany, 424 outbreaks of CSF in domestic pigs and a great number of cases in wild boar were recorded between 1990 and 1998. Most of the federal states ('Bundeslander') were affected. Epidemiological data from field investigations combined with genetic typing allowed to distinguish seven unrelated epidemics and a number of sporadic outbreaks in domestic pigs. Detailed epidemiological data was available for 327 outbreaks. It was found that 28% of these were primary outbreaks. Most of them were due to indirect or direct contact to wild boar infected with CSF virus or swill feeding. Infected wild boar remain the main risk for domestic pigs. The most frequent sources of infection in secondary or follow up outbreaks were the trade with infected pigs, neighbourhood contacts to infected farms and other contacts via contaminated persons and vehicles, respectively. An increased risk of virus transmission from infected herds to neighbourhood farms was observed up to a radius of approximately 500m. More than two thirds of the infected herds were discovered due to clinical signs. About 20% were identified by epidemiological tracing on and back. These were scrutinised because contacts to infected herds were evident. In conclusion, tracing of contact herds and clinical examination combined with carefully targeted virological testing of suspicious animals is likely to be the most important measure to immediately uncover secondary outbreaks. Obligatory serological screening in the surveillance and the restriction zones do not seem to be efficient measures to detect follow-up outbreaks. PMID- 11042399 TI - A serological survey on classical swine fever (CSF), Aujeszky's disease (AD) and porcine reproductive and respiratory syndrome (PRRS) virus infections in French wild boars from 1991 to 1998. AB - In early 1992, a CSF epizootic was clinically recognised in a wild boar population of approximately 1300 animals within an area of 250km(2) located in the east of France. In order to check the CSF situation in wild boars outside this area, a serological survey was carried out in the rest of France, for 8 consecutive years (1991-1998). This paper reports on the results obtained during this survey which included wild boars shot during the hunting period but also boars reared within fences. Around 1000-2700 sera a year were tested for the presence of antibodies to classical swine fever virus (CSFV) and also to Aujeszky's disease virus (ADV). Out of 12025 sera tested over the whole period, 80 wild boars were found positive for CSF antibodies. Sixty of them were collected on wild boars shot during the years 1992-1994 in the epizootic area located in east of France and 10 were collected in Corsica during the years 1994 1996. The last four positive samples were single reactors coming from areas or farms, which were thereafter confirmed to be serologically negative. These results together with the fact that no disease has been reported so far illustrate that the French wild boar population is probably not concerned by CSF infection (excepted in the east of France where the disease has now become enzootic). Two hundred and forty nine sera were initially detected as CSF positive but confirmed secondarily as positive for border disease (BD) antibodies. This finding shows that wild boars are also susceptible to infection by ruminant pestiviruses. Four hundred and twenty three wild boars have been found positive for ADV antibodies. In addition, from 1993 to 1995, 909 samples were tested for the presence of antibodies to porcine reproductive and respiratory syndrome virus (PRRSV). Thirty three of them were positive. The results on AD and PRRS antibody detection show that wild boars may constitute a reservoir for various infectious diseases of pigs. PMID- 11042400 TI - Genetic typing of classical swine fever virus isolates from the territory of the Czech Republic. AB - Epizootiological studies based on genetic typing were performed using 14 isolates from outbreaks of classical swine fever (CSF) in domestic pigs and wild boar in the Czech Republic which occurred between 1991 and 1998. They were compared with Austrian, Slovakian, Hungarian, Polish and German isolates. The aim of this study was to characterise the CSF virus isolates and find out the possible relationships between the outbreaks in domestic pigs and wild boar, and to map the spread of the virus in the Czech Republic. For this, fragments of the 5' nontranslated region (5' NTR) and of the E2 glycoprotein gene were sequenced and used for genetic typing. The analysis of both fragments of the genome showed that the Czech isolates belong to two CSF subgroups within group 2, namely to subgroups 2.2 and 2.3. A close relation was found with Austrian isolates from 1992 and 1994, belonging to subgroup 2.2. The isolates in subgroup 2.3 formed a very homogeneous group, although they originated from different regions of the country. They seem identical to two Slovakian isolates from 1998, and differed from Hungarian isolates from 1992. Epizootiological links became evident when the epidemiological data were compared. PMID- 11042401 TI - Classical swine fever virus: a second ring test to evaluate RT-PCR detection methods. AB - Six laboratories participated in a study to compare the sensitivity and specificity of RT-PCR tests for the detection of classical swine fever virus (CSFV). Sets of coded samples were prepared by serial dilution of positive samples and then distributed to each of the laboratories. One set comprised 25 samples of random primed cDNA, synthesised from viral RNA representative of different pestiviruses. The other set comprised samples of blood and serum obtained from virus-free or CSFV-infected pigs. Each laboratory tested the samples using PCR/RT-PCR according to a set of standardised protocols that specified the exact conditions and requirements for inclusion of control samples. Two types of test were evaluated. One amplified a part of the 5'-non coding region of the pestivirus genome by means of a closed, one-tube RT-nested PCR. The other amplified a part of the NS5B gene using non-nested RT-PCR. The results of the laboratories were compared with one another, and with those obtained earlier when similar samples were tested by the same laboratories using non-standardised methods [Paton et al., Classical swine fever virus: a ring test to evaluate RT PCR detection methods, Vet. Microbiol., in press]. Standardisation of the protocols resulted in a more consistent test sensitivity. Three laboratories avoided significant false positive results. Others that did not, could nevertheless recognise that test specificity was inadequate from the results obtained with the control samples. Minimum requirements for the inclusion of adequate controls and periodic proficiency testing are proposed. PMID- 11042402 TI - Efficacy of the classical swine fever (CSF) marker vaccine Porcilis Pesti in pregnant sows. AB - The efficacy of the classical swine fever (CSF) subunit marker vaccine Porcilis Pesti based on baculovirus expressed envelope glycoprotein E2 of CSF virus (CSFV) was evaluated in pregnant sows. Ten gilts were vaccinated with one dose of marker vaccine, followed by a second dose 4 weeks later. Four gilts remained unvaccinated and received a placebo at the same times. Thirty-three days after the second vaccination all animals were artificially inseminated. Neither local or systemic reactions nor an increase of body temperature were observed after vaccinations. All gilts showed a normal course of pregnancy. Thirty-five days after first vaccination all animals developed E2 specific neutralising antibodies with titres in the range of 5.0 and 7.5 log(2). No antibodies to CSFV-E(rns) were found in ELISA. On day 65 of gestation (126 days after the first immunisation) all sows were infected intranasally using 2ml (10(6.6) TCID(50)/ml) of the low virulent CSFV strain "Glentorf". After challenge in two of the unvaccinated control sows a slight transient increase of body temperature was observed, whereas leukopenia was demonstrated in all control animals. In addition all controls became viraemic. Vaccinations with the CSFV subunit vaccine protected the animals from clinical symptoms of CSF. In two sows a moderate decrease of leukocyte counts was detected on day 5 post infection. In contrast to the unvaccinated control sows in none of the vaccinated animals virus was isolated from the nasal swabs or the blood. Approximately 40 days after challenge all sows were killed and necropsy was done. The sows and their offspring were examined for the presence of CSFV in blood, bone marrow and different organs. No virus was found in any of the sows. In contrast, in all litters of the control sows CSFV was found in the blood as well as in the organ samples. Nine out of 10 litters of the vaccinated sows were protected from CSFV infection. Blood samples, lymphatic organs and bone marrow of these animals were all virologically negative. When sera were tested for CSFV-antibodies all sows had developed E(rns)-specific antibodies but no CSFV-specific antibodies were found in any of the progeny. It was concluded that vaccination with CSF subunit marker vaccine Porcilis((R)) Pesti protected 90% of the litters from viral infection when sows were challenged mid-gestation using the CSFV-strain "Glentorf". PMID- 11042403 TI - Safety and efficacy of a classical swine fever subunit vaccine in pregnant sows and their offspring. AB - In the study three groups with five pregnant sows each were used. The animals were vaccinated twice, 2 weeks apart, in different stages of gestation, i.e. +/ 4, +/-8 and +/-12 weeks after insemination and then 14 days later, respectively. From each group of sows three litters were randomly selected and vaccinated twice, 4 weeks apart, at 5 and 9, 7 and 11, and 9 and 13 weeks of life, respectively. Blood for serological investigations by virus neutralisation test and ELISA tests (for E(rns) antibodies and for E2 antibodies, separately) was taken before immunisation, at each vaccination and 2 weeks thereafter. Clinical observations shown that no local nor systemic reactions as well as no adverse effect on gestation occurred after vaccinations in any of the sows. Serological tests detected a low level of antibodies after the first vaccination and a typical booster effect after the second one. In piglets no adverse effect of the vaccination on the body weight gain was found. The presence of maternally derived antibodies (MDA) in non-vaccinated control piglets was observed up to the age of 5-13 weeks of life. The most evident immunological reaction was obtained in piglets vaccinated at the age of 5 or 7 weeks of life and revaccinated 4 weeks later. The CSFV-E2 subunit marker vaccine tested proved to be safe for pregnant sows and immunogenic for MDA positive piglets. PMID- 11042404 TI - Detection of classical swine fever virus in semen of infected boars. AB - During the Classical Swine Fever (CSF) epidemic in 1997 in the EU member states Germany, Italy, Spain and The Netherlands, boars in an artificial insemination (AI) centre were found to be infected with CSF virus. This raised a question of epidemiological importance which could not be answered immediately. Can CSF virus be shed by semen of infected boars and what conclusions concerning the risk of spreading CSF infection by semen can be drawn. Experimental studies were conducted to answer this question. Four young boars were infected with a CSF field virus isolate from Germany, which had been characterised in a previous animal experiment. Semen was collected at least every other day after infection. The semen was subjected to the standard diagnostic procedure for the detection of CSF virus and to semen quality assessment. The boars were euthanized at day 8, 12, 16 and 21 post infection, respectively. A post mortem examination was done and organ samples were taken from the CSF reference organs and genital organs for the detection of virus and antigen. The course of CSF infection of the boars was mild but detectable during the second week of infection. CSF virus could be isolated from semen of two animals during the pyrexic phase and from the epididymis but not from the testes. Since CSF virus shedding via semen could be proven, it was concluded that the disease may also be transmitted by artificial insemination. However analysis of semen in cell culture for the presence of CSF virus is not suitable as a routine method for CSF diagnosis. PMID- 11042405 TI - The genetic basis for cytopathogenicity of pestiviruses. AB - Two biotypes of pestiviruses, cytopathogenic (cp) and noncp viruses, can be distinguished by their effects on tissue culture cells. Identification of cp bovine viral diarrhea virus (BVDV) has been frequently reported since antigenically closely related noncp and cp BVDV can be isolated from cattle with fatal mucosal disease (MD) and are called a virus pair. In contrast to the BVDV system, only few cp border disease virus (BDV) and cp classical swine fever virus (CSFV) strains have been described. Serological analyses and sequence comparison studies showed that cp pestiviruses arise from noncp viruses by mutation. Elaborate studies during the last 10 years revealed that in most cases RNA recombination is responsible for the generation of the cp viruses. Recent results showed a second way for the development of a cp pestivirus which is based on the introduction of a set of point mutations within the NS2 gene. PMID- 11042406 TI - Cellular insertions in the NS2-3 genome region of cytopathic bovine viral diarrhoea virus (BVDV) isolates. AB - When compared to noncytopathic (ncp) bovine viral diarrhoea virus (BVDV), some cytopathic (cp) BVDV contain additional sequences in the NS2-3 genomic region. One of these insertions, which is 270 nucleotides long and of host origin (cINS), was first described for strain NADL. To find out how frequently this type of insertion occurs in other cp BVDV, 32 cp BVDV field isolates and the BVDV reference cp strain Indiana were screened using RT-PCR which detected cINS in NADL. For most cp viruses an RT-PCR product of 402bp indicated the presence of NS2-3 genes without insertions. In addition, one or two DNA fragments, around 600 850bp in size, were amplified from the genomes of 13 cp viruses indicating the presence of insertions. Sequencing of the PCR products, i.e. 402bp DNA fragment (with no insertion) and longer fragments (with insertion) revealed the location of the insertions in the NS2-3 coding region of eight cp BVDV genomes. All of the insertions were confirmed to be of the cINS type and were located in a very similar position to that found previously in the NADL genome. They were in the same reading frame as the viral polypeptide and they encoded 90-140 amino acids. The 5' and 3' ends of the insertions were different in most of the cp isolates studied. Interestingly, a 14-amino-acid stretch at the 5'-end of the insertion in the cp 5569 isolate as well as 15 amino acids at the 3'-end of the insertion in the cp 5.19516 isolate were not homologous to the cINS sequence. No significant matches for these stretches were found in the EMBL and Swissprot databases. PMID- 11042407 TI - Experiences from the Danish programme for eradication of bovine virus diarrhoea (BVD) 1994-1998 with special reference to legislation and causes of infection. AB - The main experiences from the Danish bovine virus diarrhoea (BVD) eradication programme over 5 years from 1994 to 1999 are presented. The last 3 years of the programme has been strongly supported by legislation. The most important regulations have been blood testing of live animals before movement to other herds, common pastures or exhibitions, and monitoring of all herds at regular intervals for the presence of the infection. Nevertheless, free herds have experienced infection, e.g., 204 dairy herds in 1998. Of herds found to be infected in the period from July 1997 through June 1998 after previously having been registered to be BVD-free, 67 herds were thoroughly investigated. Nineteen herds (28%) were found infected because of purchase of pregnant cows or heifers which delivered persistently infected (PI) calves, and 24 (36%) and two (3%) because of PI animals on neighbouring pastures or in neighbouring farm houses, respectively. In five herds (7%) pregnant heifers had become infected on one and the same common pasture, while in 17 herds (25%) no immediate cause of infection could be demonstrated. Yet, airborne spread from PI herds as a source of infection was suspected in some of these cases. It was furthermore concluded from investigations presented, that antibody-positive AI bulls were a remote but unlikely possibility. Free-living deer in Denmark had to be considered uninfected. Presence of PI-animals in sheep on infected farms has been seen and is paid attention to in individual cases. The results underline the need for legislation to be used in eradication programmes in areas with a high prevalence of infection and to be introduced right from the beginning in order to minimise the risk of infection for free herds. PMID- 11042408 TI - Phylogenetic, antigenic and clinical characterization of type 2 BVDV from North America. AB - Bovine viral diarrhea virus (BVDV) infection continues to have a significant impact upon US cattle producers despite the availability of more than 140 federally licensed vaccines. Detection and control is hampered by viral heterogeneity that results in differences in neutralizing epitopes, cytopathology and virulence. Recently it was found that there are two different genotypes, BVDV1 and BVDV2, among BVDV. BVDV2 isolates make up a significant proportion of the BVDV isolated in North America. Serologically BVDV2 viruses can be distinguished from BVDV1 and border disease viruses. Mab binding also distinguishes between BVDV1, BVDV2 and BDV. Like the BVDV1 viruses, BVDV2 viruses may exist as one of two biotypes, cytopathic or noncytopathic, based on their activity in cultured cells. Cytopathogenic effects on cultured cells does not correlate with virulence in vivo, as BVDV2 associated with hemorrhagic syndrome (HS) are noncytopathic. Variation among BVDV1 and BVDV2 in the 5' UTR is similar. Phylogenetic analysis and differences in virulence suggest that BVDV2 are heterogeneous. Symptoms resulting from BVDV2 infections may range from clinically inapparent to clinically severe. Recently, disease outbreaks associated with acute uncomplicated BVDV infection have been reported in the US and Canada. These outbreaks of clinically severe disease, termed HS, were all associated with viruses from the BVDV2 genotype. Not all BVDV2 isolates cause clinically severe disease. Avirulent BVDV2 isolates do exist and may predominate over virulent BVDV2 in nature. When virulent BVDV2 viruses are inoculated into calves they induce a disease characterized by fever, diarrhea, leukopenia, lymphopenia, neutropenia, thrombocytopenia, and death. Infection with avirulent BVDV2 results in a reduction of luekocytes that may be accompanied by a low-grade fever. These viruses do not cause clinical disease or a clinical leukopenia. PMID- 11042409 TI - Morphologic lesions in type 2 BVDV infections experimentally induced by strain BVDV2-1373 recovered from a field case. AB - Widespread outbreaks of severe acute BVDV, some associated with hemorrhagic syndrome (HS), were reported in Quebec and Ontario in 1993. These outbreaks caused significant economic hardship in infected herds. In the Ontario outbreak 150 dairy, 600 beef and 100 milk and grain fed veal herds were affected with losses estimated at $40000-$10000 per herd in lost animals, milk production, abortions and genetics. Fever, pneumonia, diarrhea, and sudden death occurred in all age groups of cattle. Abortions were frequently observed in pregnant cattle. The viruses associated with this outbreak were determined to be noncytopathic BVDV from the type 2 genotype. All BVDV2 associated with these outbreaks were noncytopathic. One of the viruses isolated from the Ontario outbreak, BVDV2-1373, was used to experimentally induce HS in 5-6 weeks old colostrum deprived, seronegative calves. All animals developed leukopenia and thrombocytopenia within 6-10 days with some developing bloody diarrhea and becoming moribund. Animals were killed for necropsy between 6 and 11 days postinfection. Histopathologically lesions were similar, but more severe, to those seen early on (within first 9 days after superinfection) in animals with experimentally induced mucosal disease (MD). There were no erosions and ulcerations present in the upper digestive tract. In hemorrhages in the mucosa, virus antigen (VA) was present in macrophages of both the lamina propria and the submucosa and in basal epithelial cells. Cells containing VA were vacuolated and separated from each other. The most severe lesions observed in the digestive tract were in the Peyers patches and were characterized by depletion of lymphocytes and proliferation of crypt cells resulting in crypthyperplasia. Apoptotic cells were present in crypts and areas of lymph follicles where viral antigen was detected. Out of the six animals, VA was present in four animals in the pancreas, three animals in the pituitary and in two animals in the adrenal glands. The results suggest that the pathology resulting from acute infection with a highly virulent noncytopathic BVDV2 differs from the pathology observed in classic mucosal disease. PMID- 11042410 TI - Comparative investigation of tissue alterations and distribution of BVD-viral antigen in cattle with early onset versus late onset mucosal disease. AB - Tissue alterations and distribution of BVDV antigen were examined in nine cattle with early onset and five cattle with late onset mucosal disease (MD) to evaluate the possibility to differentiate between the two disease entities. MD was induced by inoculation of persistently viremic cattle with different strains of cytopathogenic BVDV. Animals which developed early onset MD became moribund approximately 2 weeks post-inoculation (pi); animals with late onset MD 42-115 days pi. All animals were euthanized and necropsied when moribund. Macroscopic lesions were found in the upper and lower digestive tract of cattle with early and late onset MD. In cattle with late onset MD, lesions in the oral cavity were generally milder and in the intestinal tract they were not only associated with GALT, but frequently affected the mucosa outside. Histologically, the abrupt changes between hyperplastic and atrophic areas of mucosa were striking in the cattle with late onset MD. This corresponded with the multifocal distribution of areas of mucosa in which intense staining for BVD-virus antigen could be demonstrated. In both courses of MD, a severe depletion of Peyer's patches was noted, but only in late onset MD, there was a complete loss of architecture. The most distinctive difference was the presence of vascular lesions which were observed in all five cattle with late onset MD, but in none of the animals with early onset MD. The vasculopathy was characterized by segmental necrosis of vascular walls and lymphohistiocytic perivasculitis in arterioles and small arteries in the submucosa of the intestine. PMID- 11042411 TI - Clinical, pathological and antigenic aspects of bovine viral diarrhea virus (BVDV) type 2 isolates identified in Brazil. AB - Nucleotide sequencing and phylogenetic analysis of Brazilian bovine viral diarrhea virus (BVDV) field isolates identified four viruses belonging to the genotype 2. Comparison of 5' UTR sequences from these isolates to those of North American BVDV type 2 revealed genomic variations that correlated with the geographic origins of the isolates. Two of the Brazilian type 2 viruses were isolated from clinical cases of gastroenteric/respiratory disease and two were isolated from healthy bovine fetuses. The clinical cases affected young animals (8- and 18-months-old) and were characterized by diarrhea, respiratory signs, extensive oral and digestive tract erosions, conjunctival and vulvar congestion, occasional digestive bleeding and vulvar and heart petechial hemorrhage. Antigenic analysis of these isolates with a panel of 10 monoclonal antibodies revealed marked antigenic differences in the major envelope glycoprotein, gp53/E2, compared to standard laboratory and vaccine BVDV strains. In addition, virus-specific antisera raised to Brazilian BVDV type 2 viruses displayed very low serological cross-reactivity with standard BVDV type 1 strains. Differences up to 64-fold in cross-neutralization titers were observed between BVDV type 1 and Brazilian BVDV type 2 isolates. The identification of BVDV type 2 among Brazilian cattle may have important implications for epidemiological studies, diagnostic and immunization strategies. Furthermore, the low neutralizing activity of BVDV type 1 antisera against the recently identified Brazilian BVDV type 2 isolates raises the question about the degree of protection conferred by BVDV vaccines, most of them based on a single type 1 strain. PMID- 11042412 TI - Bovine viral diarrhoea virus: its effects on ovarian function in the cow. AB - Bovine viral diarrhoea virus (BVDV) is a major cattle pathogen responsible for a spectrum of symptoms, including reproductive failure. In this paper we investigate how BVDV interacts with the ovary. The viruses' tropism for the pre ovulatory oocyte was studied by indirect immunohistochemistry. Two monoclonal antibodies, raised against the non-structural protein NS3 and the envelope glycoprotein E2 were used to probe cryo-sections cut from the ovaries of three persistently infected heifers. NS3 and E2 antigens were widely distributed within the ovarian stroma and follicular cells. NS3 was also localised within the proportion of oocytes. Overall 18.7% of the oocyte population had detectable levels of NS3. What is more, the proportion of antigen positive oocytes remained constant (P>0. 05) throughout the different stages of oocyte maturation. In a subsequent study seven cows were challenged with non-cytopathogenic BVDV (strain Pe515: 5x10(6) TCID(50)) to determine the oestradiol and progesterone responses to an acute infection. The sensitivity of the endogenous luteolytic mechanism was also established by analysing plasma prostaglandin F2alpha metabolite (PGFM) levels following an exogenous oxytocin (50 IU) challenge. The inoculation was given 2 days before a synchronised oestrus and was timed to ensure that viraemia occurred during the initial stage of corpora luteal development. Seven cows inoculated with non-infectious culture medium served as control animals and remained BVDV naive throughout the study. The BVDV challenge was followed by leucopenia, viraemia and sero-conversion. The virus also significantly (P<0.01) reduced plasma oestradiol levels between day 6 and day 11 post-inoculation (i.e. between day 4 and day 9 post-oestrus). However, the infection did not alter (P>0.05) progesterone secretion throughout the oestrous cycle or the plasma concentration of PGFM. These data indicate that bovine follicular cells and oocytes are permissive to BVDV at all stages of follicular development. They also show that a transient fall in oestradiol secretion may accompany an acute infection. In conclusion, this work has identified two potential routes through which BVDV can reduce fertility in the cow, namely impairment of oocyte quality and disruption of gonadal steroidogenesis. PMID- 11042413 TI - A new inactivated BVDV genotype I and II vaccine. An immunisation and challenge study with BVDV genotype I. AB - An inactivated vaccine containing BVDV I and II strains (PT810; BVDV I, and 890; BVDV II) and using different adjuvants and antigen dosages was tested in a cattle challenge model. Groups of six healthy, seronegative cattle were vaccinated twice with a low dose (10(6.6) TCID(50) PT810 and 10(7.2) TCID(50) 890) vaccine with the adjuvant Bay R1005 or a high dose (10(7.8) TCID(50) PT810 and 10(8. 2) TCID(50) 890) vaccine with two different adjuvants (Bay R1005 or Polygen). Thirty eight days after the second vaccination, immunised animals (n=18) and non vaccinated control animals (n=3) were challenged intranasally with 10(6) TCID(50) BVDV strain PT810. For a period of 16 days, virus was isolated from blood leukocytes and nasal swabs, and neutralising antibody titres were determined.The induction of antibodies following immunisation was strongly dependent on the antigen dosage in the vaccine. The high dose formulation induced high serum neutralising antibody titres against both genotypes of up to 32000 after the second immunisation. Animals with neutralising antibody titres >512 (n=14) did not show any marked leukopenia after challenge and only very little or no virus could be isolated from blood leukocytes and/or nasal swabs when compared to control cattle. Furthermore, some of these animals did not show any boost of neutralising or even NS3-specific antibodies, which renders viral replication unlikely and thus would prevent infection of the fetus. Both adjuvants (Bay R1005 or Polygen) were similarly efficient and induced nearly identical antibody responses. In contrast, four of the six low dosage vaccinates had a marked leukopenia and viraemia as well as detectable nasal virus shedding for several days. We conclude that the selected strains and the system of vaccine preparation with high BVDV antigen dosages and highly efficient new adjuvants provide an effective means of protection against BVDV I infections. Investigations to demonstrate the protection against BVDV II infections, the duration of immunity and the ability of fetal protection by using the high dose vaccine in a fetal challenge model will follow. PMID- 11042414 TI - Development of a fetal challenge method for the evaluation of bovine viral diarrhea virus vaccines. AB - A method to evaluate the efficacy of bovine viral diarrhea virus (BVDV) vaccines using a multiple challenge model was investigated. Four pregnant heifers were challenged intranasally with a type I and type II isolate of BVDV at 75 days of gestation. At 60 days postinoculation, virus isolation and RT-PCR from blood and tissues of fetuses indicated that all fetus were persistently infected with both type I and type II isolates. Differing results of detection by PCR and virus isolation between the type I and type II isolates were obtained. These preliminary studies may indicate differences in the level of replication between type I and type II BVDV as well as predilected sites of replication in certain tissues. PMID- 11042415 TI - Translation from the internal ribosome entry site of bovine viral diarrhea virus is independent of the interaction with polypyrimidine tract-binding protein. AB - Translation of the pestiviral polyprotein is initiated cap independently at an internal site of the viral RNA, the internal ribosome entry site (IRES). We investigated the translation from the IRES of bovine viral diarrhea virus (BVDV) and the possible interaction of the unconventional cellular RNA-binding proteins, particularly of polypyrimidine tract-binding protein (PTB). The BVDV IRES is translationally active in rabbit reticulocyte lysate (RRL), and it is translated most efficiently at low concentrations of Mg(2+)- and K(+)-ions. In the UV cross link assay, several proteins from RRL bind to the BVDV IRES, including proteins of 50, 65 and 72kDa, but no protein of 57kDa possibly corresponding to PTB, although PTB is endogenously present in RRL. However, the BVDV IRES can bind PTB weakly under certain conditions. Interestingly, in a functional depletion and add back translation system, PTB does not enhance translation of BVDV, although PTB enhances translation of a picornavirus in this translation stimulation assay. These results indicate that PTB can bind the BVDV IRES RNA, but translation is independent of the action of PTB. PMID- 11042416 TI - Intranasal midazolam for prolonged convulsive seizures. AB - In order to determine the efficiency of intranasal midazolam in prolonged convulsive episodes, we conducted a prospective study in children with various types of seizures. Nine patients (six boys, three girls; age range 6 months to 9 years) with prolonged convulsions lasting more than 10 min were treated with intranasal midazolam, 0.3 mg/kg. The success rate was 100% with only one case requiring a second dose. Estimated duration of seizures was 12-30 min (mean 18.6) while mean time elapsed until cessation of seizures was 139.6 s (range 60-480). No significant adverse effects were noted except for one patient who had seizures secondary to serious CNS infection and respiratory depression after intranasal midazolam. PMID- 11042417 TI - Spatial cognition in children. I. Development of drawing-related (visuospatial and constructional) abilities in preschool and early school years. AB - The present study aimed to investigate the acquisition of visuospatial and graphomotor capacities during the pre-school and early schooling years in order to follow the normal development of drawing-related abilities and spatial cognition. Eighty children aged 3-5 years, divided in four subgroups each different for a 6-month period, and 80 children aged 8-9 years were administered a neuropsychological battery for visuospatial and visuoconstructional analysis. The battery explored five cognitive domains: visual scanning, visuospatial perceptual and representational abilities, visuomotor control and graphomotor skills. Results showed that the total scores significantly improved in each group of children with respect to the previous one, but the pattern of skill acquisition was not homogeneous. We observed a gradient from explorative and visuomotor to perceptive, representational and graphomotor abilities. Explorative and visuomotor abilities were almost mature at a time when visuoperceptual capacities began to develop. On the contrary, at that time we found very low performances at representational and constructional tasks. Our findings could suggest that constructional abilities need both perceptual and representational competences to develop properly. PMID- 11042418 TI - Spatial cognition in children. II. Visuospatial and constructional skills in developmental reading disability. AB - Cognitive models for developmental dyslexia are nowadays centered on the hypothesis of a specific deficit within the phonologic module of the language system. To ascertain whether defects of spatial cognition are associated with developmental reading disability, we investigated a sample of 43 school children (aged 8-9 years) found to be reading impaired during a wide screening survey for developmental dyslexia in the province of Naples, Italy. After one year all children were tested again and only 9/43 still presented reading impairment, while the remaining had achieved a variable range of spontaneous recovery. A detailed analysis was performed on all children to characterize their cognitive performances using on one hand classical conventional tests for constructional praxis, visuospatial cognition, and visuospatial memory and on the other a specific neuropsychological battery for constructional disorders. The results of our study demonstrated that children with long-lasting reading impairment exhibited normal performances on spatial cognition tasks. Moreover, one single child was found with relevant visuospatial deficits pointing to the possible existence of a visuospatial subtype for developmental dyslexia. PMID- 11042419 TI - Combined therapy with hypothermia and anticytokine agents in influenza A encephalopathy. AB - Two children with influenza A-related encephalopathy were treated with a combination of mild hypothermia (deep body temperature of the forehead: 35 degrees C) and anticytokine agents (high-dose methylprednisolone and ulinastatin), while receiving amantadine. One of the cases exhibited acute necrotizing encephalopathy on computed tomography (CT). Although no severe complications occurred, correctable hypokalemia and hyperglycemia occurred in both cases. Both patients recovered without any neurological sequelae. Our therapeutic protocol appears to be effective for managing influenza A-related encephalopathy. PMID- 11042420 TI - The test of variables of attention (TOVA) is useful in the diagnosis of Japanese male children with attention deficit hyperactivity disorder. AB - The purpose of this study was to evaluate the ability of the test of variables of attention (TOVA) to distinguish between 6- to 12-year-old Japanese male children with attention deficit hyperactivity disorder (ADHD group; n=17) meeting DSM-IV and ICD-10 criteria, and age-matched, normal Japanese male controls (control group; n=19). The TOVA is a computer-administered, visual continuous performance test that provides measures of attention. The ADHD group had significantly higher means than the control group in all variables: omission errors, commission errors, response time, response time variability, anticipatory responses, and multiple response. Control children exhibited age-related changes in two variables: response time and response-time variability, but no age-related changes were observed in any variables in the ADHD group. This preliminary study indicates that the TOVA makes a useful contribution to the diagnosis of Japanese male children with ADHD. PMID- 11042421 TI - Adenylosuccinase deficiency: an unusual cause of early-onset epilepsy associated with acquired microcephaly. AB - Adenylosuccinase deficiency, an autosomal recessive inborn error of purine synthesis, was first described in 1984 by Jaeken and Van den Berghe (reviewed in J Inher Metab Dis 20;1997:193). The cardinal features are variable psychomotor delay often accompanied by epilepsy and autistic features. Diagnosis is made by detection of abnormal purine metabolites in body fluids. We report a girl who presented with early onset epilepsy, associated with acquired microcephaly and severe psychomotor retardation, as the most prominent symptoms. PMID- 11042422 TI - Partial seizures in leukoencephalopathy with swelling and a discrepantly mild clinical course. AB - We report a patient with 'Leukoencephalopathy with swelling and a discrepantly mild clinical course', an entity of leukoencephalopathy recently clarified. Our patient presented with complex partial seizures in addition to characteristic radiological findings and clinical course. A review of the literature revealed that this new neurodegenerative disease complicates epilepsy in more than half of the patients, and that partial components in the seizure symptomatology are not infrequent. PMID- 11042423 TI - Sneddon syndrome, arylsulfatase A pseudodeficiency and impairment of cerebral white matter. AB - We describe a 11 year-old-boy with Sneddon syndrome, confirmed by skin biopsy, and MR evidence of diffuse cerebral hyperintensity of white matter; he also suffered from pre-perinatal hypoxic-ischemic distress. Arylsulfatase A activity was found reduced because of arylsulfatase A pseudodeficiency. We suggest that the association of pre-perinatal distress, Sneddon syndrome and arylsulfatase A pseudodeficiency is responsible for the diffuse impairment of cerebral white matter, never reported in Sneddon syndrome and similar to described cases of delayed posthypoxic demyelination and arylsulfatase A pseudodeficiency. PMID- 11042424 TI - An autopsy case of atypical adrenoleukomyeloneuropathy in childhood. AB - Adrenoleukomyeloneuropathy (ALMN) usually occurs in adulthood, it being extremely rare in childhood. We reported a quite atypical clinical case of ALMN as a variant of adrenoleukodystrophy (ALD). The onset was at 5 years 7 months and ataxia was the major symptom. His condition progressed rapidly to a vegetative state within 1 year. At the age of 11 years and 11 months he died of pneumonia and an autopsy was performed. We herein reported the neuropathological findings in this rare case. The autopsy revealed marked atrophy with diffuse demyelination and astrogliosis throughout the cerebrum, cerebellum and brainstem. Massive degeneration of the pyramidal tracts and loss of neurons were also seen in the spinal cord. The adrenal cortex showed marked atrophy with a striated cytoplasm in ballooned cells. These findings include pathological characteristics of both ALD and adrenomyeloneuropathy (AMN), suggesting ALMN. However, diffuse demyelination with gliosis in the cerebrum and cerebellum is quite atypical for ALMN. They might explain his atypical clinical course, especially the early onset of the disease with ataxia and rapid deterioration. PMID- 11042425 TI - Ictal increased writing preceded by dysphasic seizures. AB - Ictal increased writing has never been concretely demonstrated to be an epileptic symptom. We reported the evolution of seizures, from the beginning of reading writing dysphasia through increased writing to secondary generalization, in a schoolchild with benign partial seizures. The lateralizing value of inter- or non ictal hypergraphia has been stressed for the right hemisphere, while ictal increased writing most likely originated from the language area in the dominant (left) cerebral hemisphere in our patient. PMID- 11042426 TI - Education and counseling approaches in chronic conditions. PMID- 11042427 TI - Effects on quality of care for patients with NIDDM or COPD when the specialised nurse has a central role: a literature review. AB - Chronic care has to be organised in a way that care from any one caregiver is linked up to that provided by others so that disturbing gaps, contradictions and overlaps are avoided. In the search for the most effective and efficient combination of health professionals to deliver care to chronic patients, the role of the specialised nurse has become important. This article reviews a Medline search for publications about the effects of models of care for patients with NIDDM or COPD in which the specialised nurse has a central role. Main features of the models are identified and related to expected and statistically significant effects. In this young domain of effect evaluation ten publications met our criteria. Depending on the division of tasks between care providers, improvements are seen in self-care, quality of life and patient satisfaction, as well as increased medical consumption. More methodologically suitable evaluations with the use of only valid measures are needed. PMID- 11042428 TI - Fear of hypoglycaemia among insulin-treated Hong Kong Chinese patients: implications for diabetes patient education. AB - This study investigated fear of hypoglycaemia among insulin-treated Chinese patients in Hong Kong. The study employed a cross-sectional descriptive design. Using structured-interviews, the authors collected data from 120 insulin-treated adults attending a diabetes centre. Results showed that although hypoglycaemic attacks were generally mild, they were common amongst one-third of patients. Fear was predominantly low, however 15% of respondents reported high fear. In addition, fear correlated positively with time since insulin-treatment, frequency of hospitalization due to hypoglycaemia, and frequency of hypoglycaemia affecting working life. Results also showed that although all respondents were insulin treated, 40% had undesirable metabolic control. No statistically significant differences were found between the respondents' rating of adequacy of their diabetes knowledge among three groups of respondents who had completed, defaulted or were attending an education programme. Implications for diabetes patient education and further study are discussed. PMID- 11042429 TI - Health care seeking behaviour of patients with sexually transmitted diseases: determinants of delay behaviour. AB - The study aimed to determine the factors associated with a delay in seeking health care for symptoms of sexually transmitted diseases (STDs) among a sample of 1482 patients attending STD clinics. Of the sample 78% were male, 58% sought help from the clinic within the first 6 days of noticing symptoms, 24% waited between 7 and 10 days and 17% waited longer than 10 days before seeking health care. Patients who delayed were those who treated themselves prior to seeking health care, who were female, whose friends waited before seeking treatment, who held misconceptions regarding the cause of STDs, who perceived STDs not to be serious and who valued personal autonomy in sexual behaviours less and had less positive outcome expectations of refusing sex. The data suggest that targeted interventions should be directed specifically at women and the youth. Early health care seeking could be facilitated through improved basic knowledge regarding STDs, control over one's own sexual behaviour and social support for early health care seeking. PMID- 11042430 TI - Stroke patients' needs and experiences regarding autonomy at discharge from nursing home. AB - In this qualitative study stroke patients rehabilitating in nursing homes experienced an increase in their autonomy (particularly in self-determination, independence and self-care) in the last weeks before discharge. The change in autonomy was found to be related to regained abilities and self-confidence, and to patients' strategies (e.g. taking initiative, being assertive). The attitude of health professionals and family, and the nursing home could influence patient autonomy. Overprotection, paternalism, care routines and an inconsistent approach constrain autonomy. Conversely, attentiveness, tailored interventions and a respectful dialogue facilitate autonomy, like moderate instrumental and emotional support by the family. Nursing homes can enhance autonomy by minimizing care routines and by providing room for doing activities independently and privately. Attention to patient autonomy may improve patients' active participation in rehabilitation, quality of life, and autonomous living after discharge. Multidisciplinary guidelines based on the results may increase attention to the stroke patients' autonomy and stimulate a team approach. PMID- 11042431 TI - Late diagnosis of congenital hearing impairment in children: the parents' experiences and opinions. AB - The purpose of the present investigation is to describe how parents experience a delayed identification of their child's hearing impairment. Ten parents of 8 children were interviewed. The impairment was confirmed when the children were between 2 years, and 5 years and 8 months. The results show that the parents and their child pass through a series of distinct phases: Unawareness, Suspicion, Confirmation and Habilitation. After the birth of the child there was first a calm period, which lasted until the possibility of a hearing impairment was suspected. Once the suspicion was raised, a time of much anxiety and frustration ensued. The parents described how defective communication and misunderstanding lead to frequent conflicts with their child. The differing behaviour of the child, in combination with poor language development, initiated referral to audiological assessment and confirmation of the hearing impairment. After confirmation, the parents felt relief but at the same time a sorrow. When hearing aids had been fitted and education in sign language was under way, the child's language and social behaviour improved. Supposedly, the late detection is explained by the combination of an insufficient test method that cannot detect all children with a hearing impairment and, in cases of uncertainty, a tendency to let the child pass rather than "bringing bad news". All parents in the present study would have wished to participate in a hearing screening program for new borns, had the opportunity been present. PMID- 11042432 TI - Personal accounts of stroke experiences. AB - As there appeared to be a need for personal accounts of stroke experiences, a book called "Speaking about Stroke" was written for stroke patients and their caregivers. For the past two years, a questionnaire was sent to the people who had ordered the book, to gain an insight into the characteristics of those who bought it, the distribution, use, and relevance of the book, and its effects on the readers' attitude and behaviour. It appears that healthy caregivers of relatively young but severely handicapped stroke patients order the book. The book is read carefully. Both stroke patients and caregivers value the book because the accounts give them a feeling of recognition, appreciation and understanding. Many readers state that they have put suggestions given in the book into practice. "Speaking about Stroke" fills a clear need for personal accounts, and cerebrovascular accident professionals should draw the attention of the target group to it. PMID- 11042433 TI - Using content analysis of video-recorded consultations to identify smokers' "readiness" and "resistance" towards stopping smoking. AB - Although physicians' brief advice against smoking is effective in helping smokers to stop, very little is known about the process of counseling smokers. We describe the development of a coding system for describing smokers' motivation to stop smoking as judged by their behaviour when discussing smoking with their family physician. We analysed video-recordings of consultations between 47 self reported smokers and 29 family physicians where smoking was discussed. By a process of observing recordings, reading transcripts and discussion, we developed a coding system for describing smokers' levels of motivation to stop smoking. This consists of four behaviours thought to indicate "readiness" (higher motivation to stop) and four "resistance" (lower motivation to stop). Interobserver reliability between two observers was good (Kappa = 0.71 for readiness, 0.73 for resistance behaviours). We discuss the relevance of our descriptions for clinicians and the possibility of developing this work further to enable systematic research into doctor-patient interactions where smokers are counseled to stop. PMID- 11042434 TI - Infertility counselling: the ISSUE experience of setting up a telephone counselling service. AB - This paper explores the role of telephone counselling for people experiencing fertility problems. It examines the first nine months of the evening telephone counselling service offered by ISSUE (The National Fertility Association). Records of the service were analysed for number, duration and nature of calls. The results suggest that there is significant demand for such a service and that this demand has grown in line with availability. All counsellors working on the service were interviewed. Qualitative analysis of the interviews generated key themes concerning gender and culture, supervision and training, counselling process issues, boundaries and managerial issues. In addition, specific issues about counselling in the field of reproductive medicine were highlighted. Findings are discussed in relation to implications for service provision in this area of health care. PMID- 11042435 TI - The role of cognition in vocational functioning in schizophrenia. AB - Schizophrenia is associated with long-term unemployment. Cognitive dysfunction, rather than clinical symptoms, may be the most important factor in the ability to work for patients with this disorder. To evaluate the relationship of clinical symptoms and cognitive functioning to work status, thirty patients with schizophrenia, who were participants in a vocational rehabilitation program, were evaluated with a comprehensive neuropsychological battery and assessment of psychopathology. Subjects were classified as being in stable full-time, part-time or unemployed work status for at least a year. Univariate analysis indicated that patients who were working full-time were significantly better educated, more likely to be treatment-resistant, more likely to be treated with an atypical antipsychotic medication, had more positive symptoms, and were engaged in work tasks which were more cognitively complex than the part-time employed and unemployed work groups. An ANCOVA controlling for education demonstrated that the full-time employed group performed significantly better than the unemployed group on measures of executive functioning, working memory and vigilance; and significantly better than the part-time group on measures of vigilance and executive functioning. Although negative symptoms did not significantly relate to work status in the univariate analysis, a multiple regression indicated that negative symptoms, level of education, and executive functioning differentiated the work groups. These results suggest that poor premorbid function, negative symptoms and cognitive dysfunction are significantly associated with unemployment in schizophrenia. PMID- 11042436 TI - Gender differences in premorbid cognitive performance in a national cohort of schizophrenic patients. AB - Despite significant research, there are still inconsistent findings regarding gender differences in cognitive performance in individuals already diagnosed with schizophrenia; studies have found that males suffering from schizophrenia are more, less or equally impaired compared with females. Gender differences in cognitive performance in individuals suffering from schizophrenia may be influenced by gender differences in premorbid cognitive performance; the very few and very small N studies published indicated that males have a poorer pre-morbid cognitive performance than females. This study examined the gender differences in premorbid cognition, utilizing cognitive assessments performed on female and male adolescents before induction into military service. The Israeli Draft Board Registry, which contains cognitive assessments equivalent to IQ scores on 16-18 year old Israeli adolescents, was linked with the Israeli National Psychiatric Hospitalization Case Registry, which records all psychiatric hospitalizations in the country. Scores on premorbid cognitive performance in schizophrenia were examined in 90 female-male case pairs matched for school attended as a proxy for socio-economic status. The mean age of first hospitalization was 20. 1+/-1.8 years of age for males and 19.6+/-1.8 years of age for females. A repeated measures ANCOVA with age of first hospitalization and years of formal education as covariates, and controlling for gender differences in cognitive performance in healthy adolescents, revealed a significant difference in pre-morbid cognitive performance between males and females on all four cognitive measures [F(1,87)=8.07, P=0.006] with females scoring lower (worse) than males. In this national cohort, pre-morbid cognition was poorer in female, compared with male, adolescents who will suffer from schizophrenia in the future, a result consistent with some, but not all, similar studies. These results may be valid only for patients with first hospitalization around age 20. Hence, gender differences in premorbid cognition should be taken into account when assessing gender differences in cognition in schizophrenia. PMID- 11042437 TI - Phasic and enduring negative symptoms in schizophrenia: biological markers and relationship to outcome. AB - Negative symptoms have been associated with poor response to neuroleptics, enlarged ventricles, cognitive impairment, and poor outcome in schizophrenia. These associations appear, however, to be dependent on the phase of study, suggesting that acute-phase (phasic) negative symptoms may be pathophysiologically distinct from enduring negative symptoms that persist through the residual phase. To compare correlates of enduring and phasic negative symptoms, we studied 60 drug-free schizophrenic patients (DSM-III-R and SADS/RDC) at baseline, 4 weeks after neuroleptic treatment, and assessed the 1 year outcome. We rated positive and negative symptoms at baseline and 4 weeks after treatment. At baseline, premorbid function, neuropsychological function, ventricle-brain ratio (VBR) and symptom response to an anticholinergic agent were assessed, and a two-night sleep EEG and 1mg dexamethasone suppression test (DST) were conducted. Phasic negative symptoms were defined as the change in negative symptoms (baseline to 4 weeks) and enduring negative symptoms as severity of negative symptoms at 4 weeks. Patients had varying proportions of phasic and enduring symptoms; the two did not define distinct subgroups. Phasic negative symptoms were significantly correlated with global treatment response, positive symptom treatment response, response to anticholinergic agent, baseline post dexamethasone cortisol, and shortened REM latency. Enduring negative symptoms were significantly correlated with residual positive symptoms and global psychopathology, VBR, poor performance on neuropsychological testing, decreased slow-wave sleep, poor premorbid function, and poor 1 year outcome. These data suggest that phasic negative symptoms and enduring negative symptoms may be caused by different pathophysiological mechanisms. PMID- 11042438 TI - Self-Appraisal of Illness Questionnaire (SAIQ): relationship to researcher-rated insight and neuropsychological function in schizophrenia. AB - The Self-Appraisal of Illness Questionnaire (SAIQ) is a self-report instrument designed to assess attitudes toward mental illness among persons receiving psychiatric treatment. This instrument was developed for use in community settings, adapted closely from the Patient's Experience of Hospitalization questionnaire. In order to examine the validity of the SAIQ, a factor analysis was first conducted on the items of this instrument in a sample of 59 outpatients with schizophrenia or schizoaffective illness. Three factors emerged: Need for Treatment, Worry, and Presence/Outcome of Illness. Next, to examine the concurrent validity of these three SAIQ subscales, they were correlated with researcher rated insight scales and neuropsychological tests. Results indicated that the Need for Treatment and Presence/Outcome subscales were significantly correlated with both researcher-rated insight scales and with neuropsychological tests of executive functioning. The Worry subscale was not associated with either researcher-rated insight scales or neuropsychological tests. It was concluded that the Need for Treatment and Presence/Outcome subscales may be used in combination as a brief screening instrument for clients with schizophrenia receiving outpatient psychiatric treatment who may be at risk for treatment non compliance due to a lack of insight into illness. PMID- 11042439 TI - The kings schizotypy questionnaire as a quantitative measure of schizophrenia liability. AB - We used a new self-report measure, the Kings Schizotypy Questionnaire (KSQ; Williams, M. The psychometric assessment of schizotypal personality. PhD thesis. Institute of Psychiatry, University of London, 1993), to investigate schizotypy as a quantitative measure of familial liability to schizophrenia. The KSQ was administered to 135 DSM-IV schizophrenia probands, 153 of their healthy first degree relatives, and 267 control subjects. We found that the questionnaire clearly differentiated schizophrenic from non-schizophrenic individuals, but failed to differentiate the relatives from controls. Possible reasons for this include defensive responding among relatives, self-selection bias among relatives, differences in data collection methods, and the possibility that positive aspects of schizotypy may not be closely related to familial liability to schizophrenia. PMID- 11042440 TI - Attitudes towards neuroleptic treatment: reliability and validity of the attitudes towards neuroleptic treatment (ANT) questionnaire. AB - Non-compliance problems may rise to 50% among patients undergoing neuroleptic treatment. There are no direct measures available to predict compliance, if previous non-compliance is not taken into account. Attitudes towards neuroleptic treatment and insight into psychotic symptoms may vary during the course of the treatment process. It would be relevant to evaluate these items before taking any clinical action and later reassess the degree of change. The instrument thus far available has been the Drug Attitude Inventory. It has limitations for use with first-episode-patients and their follow-up. Its statements are dichotomous, which makes it difficult to determine the variation of attitudes e.g. during maintenance treatment, and most of the items concentrate on the subjective state of the patient, leaving attitudes as a minority in the scale. In this study, we report the new Attitudes towards Neuroleptic Treatment (ANT) questionnaire for the quantitative assessment of attitudes. We developed 10 statements for attitudes and two items for insight in the Visual Analogue Scale form (0-100 points). These were compared with the Drug Attitude Inventory 10 Questionnaire (DAI-10) (Hogan, T. P., Awad, A.G., Eastwood, R., 1983. A self-report scale predictive of drug compliance in schizophrenics: reliability and discriminative validity. Psychol. Med. 13, 177-183.) among 106 subjects receiving neuroleptic medication. The 12 Visual Analogue Scales showed a high inter-item consistency and fair test-retest validity. The results were in accordance with the DAI-10. The scales comprised three factors: general attitudes, subjective feeling and expectations and insight. Attitudes towards neuroleptic treatment and insight into psychotic symptoms are different dimensions and can be measured quantitatively. The Attitudes towards Neuroleptic Treatment scale is useful in assessing the state of attitudes before starting medication and for follow-up among patients receiving neuroleptic medication. PMID- 11042441 TI - Saccadic eye movement abnormalities in relatives of patients with schizophrenia. AB - Recent studies note abnormalities in saccadic eye movements of relatives of patients with schizophrenia. The current study examined which aspects of the saccadic system are affected, whether these saccadic abnormalities are associated with schizophrenia spectrum personality symptoms (SSP), and whether such an association is dependent on a family history of schizophrenia. Furthermore, the study examined what proportion of relatives have the saccadic abnormality(ies). Fifty-five first-degree relatives with no DSM-III-R Axis I diagnosis participated in the study. Twenty-one of these relatives experienced SSP symptoms and 34 had no Axis II diagnosis. Sixty-two subjects with no Axis I diagnosis were recruited from the community. Twenty-five experienced SSP symptoms and 37 had no Axis II diagnosis. Prosaccades (saccades toward the target) and antisaccades (saccades made in the opposite direction of the target jump) were examined. Relatives, particularly those with SSP, had difficulties with the antisaccade task as suggested by higher error rates and longer antisaccade latency. Prosaccades were not different in relatives compared to the community subjects, although the effects of field were different in the two groups on some measures. The antisaccade latency was 'abnormal' in only a small proportion (1.6%) of community subjects compared to 14.9% of all relatives (35.3% of SSP relatives and 3.3% of non-SSP relatives). Relatives of patients with schizophrenia have deficits in aspects of the saccadic system involved in generating internally driven saccades and inhibition of unwanted saccades. These deficits implicate frontal ocular motor neuronal circuitry involving frontal cortical and basal ganglia areas. These deficits are associated with SSP symptoms, but not in the absence of a blood relationship to schizophrenia. The relatively high prevalence rate of the abnormality in at-risk subjects may have relevance for use of these measures in linkage analysis. PMID- 11042442 TI - Validation and factorial structure of a standardized neurological examination assessing neurological soft signs in schizophrenia. AB - Although neurological soft signs (NSS) have been consistently reported in patients with schizophrenia, their clinical relevance, the actual impact of treatment or their evolution during the disease are not well clarified, possibly because of methodological limitations of the available tools. We have developed a new standardized examination integrating the assessment of 23 NSS selected from the literature and the rating of well-validated scales for assessment of extra pyramidal symptoms. We examined 161 subjects (controls, n=48; patients with schizophrenia, n=95; or recurrent mood disorder, n=18). Half of the patients were neuroleptic-free. Schizophrenic patients had significantly higher total score (14. 6+/-8) than mood disorder patients (12.0+/-7) and controls (5.0+/-2). Internal consistency (Cronbach's alpha=0.85) and inter-rater reliability were good. Principal component analysis found five consistent factors ('motor coordination', 'motor integrative function', 'sensory integration', 'involuntary movements or posture', 'quality of lateralization'). This scale thus confirmed a factorial structure in agreement with the conceptual areas of interest explored by NSS and should be a useful tool for assessment of the different dimensions of neurological dysfunction in schizophrenia. PMID- 11042443 TI - Altered brain activation in schizophrenia during visually guided motor selection revealed by fMRI. PMID- 11042444 TI - Erratum to "New strategies for old problems: tardive dyskinesia (TD). Review and report on severe TD cases treated with clozapine, with 12, 8 and 5 years of video follow-up" PMID- 11042445 TI - Carbohydrates and glycoconjugates. Biophysical methods. Web alert. PMID- 11042446 TI - Glycosyltransferases, sugar nucleotide transporters and bacterial surface lectins - at the cutting edge of glycobiology PMID- 11042447 TI - Glycosyltransferase structure and mechanism. AB - The high-resolution X-ray crystal structures of a new form of bacteriophage T4 beta-glucosyltransferase, Escherichia coli MurG, Bacillus subtilis SpsA, bovine beta-1,4-galactosyltransferase 1 and rabbit N-acetylglucosaminyltransferase I have now been solved. These glycosyltransferase structures have provided the first detailed view of the structural basis of catalysis, as well as new insight into glycosyltransferase classification. PMID- 11042448 TI - Recent advances in the study of the biosynthesis and functions of sulfated glycosaminoglycans. AB - Recent cDNA cloning of the glycosyltransferases involved in the synthesis of the sulfated glycosaminoglycan sidechains of proteoglycans has provided important clues to answering long-standing questions concerning the mechanisms of both chain polymerization and the biosynthetic sorting of glucosaminoglycans (heparin/heparan sulfate) and galactosaminoglycans (chondroitin/dermatan sulfate). These biosynthetic mechanisms are crucial to the expression and regulation of the biological functions of glycosaminoglycans in development and pathophysiology. PMID- 11042449 TI - Microbial toxins and the glycosylation of rho family GTPases. AB - Large clostridial cytotoxins act on cells by glycosylating low molecular mass GTPases using nucleotide-sugars as the sugar donor. These toxins are important virulence factors in human and animal diseases, but are also valuable cell biology tools. Recent findings shed some light on their mode of action and provide new insights into the structure/activity relationship of these bacterial toxins. PMID- 11042450 TI - Large-scale production of oligosaccharides using engineered bacteria. AB - Rapid advances in the cloning and expression of glycosyltransferase genes, especially from bacteria, could open the way to overcoming difficulties in the mass production of oligosaccharides. The large-scale production of oligosaccharides using either glycosyltransferases isolated from engineered microorganisms or whole cells as an enzyme source could promote a new era in the field of carbohydrate synthesis. PMID- 11042451 TI - Nucleotide sugar transporters of the Golgi apparatus. AB - Glycosylation, sulfation and phosphorylation of proteins, proteoglycans and lipids occur in the lumen of the Golgi apparatus. The nucleotide substrates of these reactions must be first transported from the cytosol into the Golgi lumen by specific transporters. The topology and structure of these hydrophobic, multi transmembrane-spanning proteins are beginning to be understood. PMID- 11042453 TI - New approaches to study macromolecular structure and function PMID- 11042454 TI - High-throughput protein crystallization. AB - The combinatorial chemistry industry has made major advances in the handling and mixing of small volumes, and in the development of robust liquid-handling systems. In addition, developments have been made in the area of material handling for the high-throughput drug screening and combinatorial chemistry fields. Lastly, improvements in beamline optics at synchrotron sources have enabled the use of flash-frozen micron-sized (10-50 microm) crystals. The combination of these and other recent advances will make high-throughput protein crystallography possible. Further advances in high-throughput methods of protein crystallography will require application of the above developments and the accumulation of success/failure data in a more systematic manner. Major changes in crystallography technology will emerge based on the data collected by first generation high-throughput systems. PMID- 11042452 TI - Chaperone-assisted pilus assembly and bacterial attachment. AB - Bacterial pili assembled by the chaperone-usher pathway can mediate microbial attachment, an early step in the establishment of an infection, by binding specifically to sugars present in host tissues. Recent work has begun to reveal the structural basis both of chaperone function in the biogenesis of these pili and of bacterial attachment. PMID- 11042455 TI - Recent developments in software for the automation of crystallographic macromolecular structure determination. AB - The automation of macromolecular structure determination by X-ray crystallography has long been a goal for many researchers. Recently, there have been improvements in the underlying algorithms, some of which have been implemented in software packages that deal with multiple stages of the structure determination process. These first steps towards complete automation have made X-ray crystallography more efficient. PMID- 11042456 TI - Single-molecule X-ray diffraction. AB - Free-electron lasers could provide femtosecond X-ray flashes with a peak brilliance 10-11 orders of magnitude higher than that which is currently available from synchrotrons. Such pulses may allow structural studies of single biomolecules before radiation damage destroys them and may permit the imaging of complex structures without the need to amplify scattered radiation through Bragg reflections. PMID- 11042457 TI - The current excitement in bioinformatics-analysis of whole-genome expression data: how does it relate to protein structure and function? AB - Whole-genome expression profiles provide a rich new data-trove for bioinformatics. Initial analyses of the profiles have included clustering and cross-referencing to 'external' information on protein structure and function. Expression profile clusters do relate to protein function, but the correlation is not perfect, with the discrepancies partially resulting from the difficulty in consistently defining function. Other attributes of proteins can also be related to expression-in particular, structure and localization-and sometimes show a clearer relationship than function. PMID- 11042458 TI - New developments in isotope labeling strategies for protein solution NMR spectroscopy. AB - The development of novel isotope labeling strategies for proteins has facilitated the study of the structure and dynamics of these molecules. In addition, the recent emergence of alternative methods of bacterial expression for obtaining isotopically labeled proteins permits the study of new classes of important proteins by solution NMR methods. PMID- 11042459 TI - Solid-state NMR spectroscopy applied to membrane proteins. AB - One major remaining problem in structural biology is to elucidate the structure and mechanism of function of membrane proteins. On the basis of preliminary information from genome projects, it is now estimated that up to 50,000 different membrane proteins may exist in the human being and that virtually every life process proceeds, sooner or later, through a membrane protein. Solid-state NMR spectroscopy in high magnetic field is rapidly developing into a widely applicable tool to describe the structure and help understand the mechanism of function of a membrane protein. Recent work in applied solid-state NMR spectroscopy crosses the boundary between the biological and the physical sciences, and aims at increasing the predictive range of this biophysical method. PMID- 11042460 TI - Protein complexes and analysis of their assembly by mass spectrometry. AB - The utility of mass spectrometry for the analysis of proteins has grown enormously in the past decade. Significant advances in detection and ionization techniques are allowing questions about noncovalent assembly to be addressed by the direct observation of gas phase complexes, their assembly in real time and their disassembly by perturbation of solution or instrument conditions. These technological innovations have plainly captured the imagination of biological researchers. Recent and novel developments include the combination of mass spectrometry with isotopic labeling, affinity labeling and genomic information. Collectively, these advances are opening new doors to the isolation of complexes, the identification of their substituents and the characterization of their conformations and assembly. PMID- 11042461 TI - Protein machines and lipid assemblies: current views of cell membrane fusion. AB - Protein machines and lipid bilayers both play central roles in cell membrane fusion, a process crucial to life. Recent results provide clues to how both components function in fusion. Recent observations suggest a common mechanism by which very different fusion machines (from lipid-enveloped viruses and synaptic vesicles) may function to produce compartment-joining pores. This mechanism presumes that fusion proteins act as machines that use stored conformational energy to assemble closely juxtaposed lipid bilayers, bend these to form fusion competent structures, stabilize unfavorable lipid structures and destabilize a committed intermediate to drive fusion pore formation. PMID- 11042462 TI - Recent developments in cryo-electron microscopy reconstruction of single particles. AB - Cryo-electron microscopy and single-particle 3D image reconstruction techniques have been used to examine a broad spectrum of samples ranging from 500 kDa protein complexes to large subcellular organelles. The attainable resolution has improved rapidly over the past few years. Structures of both symmetric and asymmetric assemblies at approximately 7.5 A have been reported. Together with X ray crystallography, three-dimensional cryo-electron microscopy reconstruction has provided important insights into the function of many biological systems in their native biochemical contexts. PMID- 11042463 TI - New perspectives in diabetes research and treatment. PMID- 11042464 TI - Free fatty acids and pathogenesis of type 2 diabetes mellitus. AB - Plasma free fatty acids (FFA) might mediate the insulin resistance and impaired glucose tolerance associated with central obesity. Central adipocytes are resistant to insulin, suggesting that FFA delivery to the liver via the portal vein is increased when visceral triglyceride (TG) stores are increased. Muscle insulin resistance might result from the 'Randle' mechanism, from downregulation of the insulin signaling pathway, and/or reduced access of insulin to skeletal muscle owing to changes in blood flow or insulin transport across capillary endothelium. TG storage within muscle might interfere with insulin action, but a causal relationship between myocellular lipid and glucose disposal remains to be demonstrated. Basal levels of FFA appear to be permissive for insulin secretion; however, elevated FFA have a minor effect on insulin secretion in vivo. In humans, prolonged hyperlipidemia engenders an insulin response matched to the degree of insulin resistance, leaving open the question of whether lipotoxicity of islet cells contributes to glucose intolerance and diabetes in humans. Elevated portal FFA might account for overproduction of liver glucose output with visceral adiposity. Additionally, portal FFA might reduce hepatic extraction of insulin, diminishing the necessity of increased beta-cell response to compensate for FFA-driven insulin resistance. Overall, effects of FFA can lead to several components of the insulin resistance syndrome and risk for diabetes. Reduction in FFA might be the appropriate therapy for these disorders. PMID- 11042465 TI - GLUT4 and company: SNAREing roles in insulin-regulated glucose uptake. AB - The primary physiological role of insulin is in glucose homeostasis. This is accomplished through the inhibition of gluconeogenesis in the liver and the stimulation of glucose uptake into insulin-sensitive tissues, such as adipose tissue, skeletal muscle and cardiac muscle. The ability of insulin to stimulate glucose uptake relies on a complex signaling cascade that leads to the translocation of glucose transporter protein 4 (GLUT4) from an intracellular compartment to the plasma membrane, which results in increased glucose uptake. Defects in the ability of insulin to regulate this key metabolic event can lead to insulin resistance and non-insulin-dependent type 2 diabetes mellitus (T2DM). To design effective treatments for diabetes, there have been major efforts to understand the insulin-regulated mechanisms that govern glucose uptake. These have involved defining the components of the insulin signaling network and identifying the molecular machinery that is used to translocate GLUT4. PMID- 11042466 TI - PPAR gamma and the treatment of insulin resistance. AB - Numerous studies across several population groups have indicated that insulin resistance plays a central role in the development of type 2 diabetes mellitus (T2DM). Moreover, this disorder is also strongly associated with other metabolic syndromes, including hypertension, dyslipidemias and polycystic ovarian syndrome (PCOS). Recent advances have demonstrated that pharmacological agents of the thiazolidinedione class can reverse insulin resistance and profoundly improve many of these associated symptoms. These effects have been documented in a variety of genetic and acquired animal models of insulin resistance, as well as in numerous clinical trials in patients with insulin resistance. These compounds appear to enhance insulin action by modulating the activity of the nuclear receptor peroxisome proliferator-activated receptor (PPAR) gamma. This activation results in changes in the expression of a number of genes that are critically involved in glucose and lipid metabolism, as well as in insulin signal transduction. While precise events that occur downstream from PPAR gamma modulation remain uncertain, new insights are emerging from knockout studies in mice and the identification of genetic variants in humans. These findings indicate that there is still much to learn about the molecular biology and physiology of these interesting receptors, and that research in this area can lead to more effective and safer drugs to treat insulin resistance and associated syndromes. PMID- 11042467 TI - RAGE: a new target for the prevention and treatment of the vascular and inflammatory complications of diabetes. AB - Although the underlying causes of hyperglycemia are multiple, a common thread associated with high levels of blood sugar is the development of a range of vascular and inflammatory complications that might seriously limit the quality and duration of life in affected individuals. Despite multiple aggressive efforts to prevent complications, diabetes remains the leading disease consuming healthcare dollars in the USA. This review focuses on the role of advanced glycation endproducts (AGEs) and their interaction with their signal-transduction AGE receptor (RAGE), in vascular and inflammatory cell perturbation and the chronic activation that underlies diabetes. Our studies provide mechanistic insights into complications within the macrovasculature and those ensuing from an exaggerated host response to invading bacteria, and suggest that blockade of RAGE might provide a potent and safe strategy for the prevention of complications that typify long-term diabetes. PMID- 11042468 TI - Life and death of the pancreatic beta cells. AB - Pancreatic beta cells are responsible for maintaining the body's glucose levels within a very narrow range; their population is dynamic, with compensatory changes to maintain euglycemia. Throughout the lifetime of a mammal, low levels of beta-cell replication and apoptosis are balanced and result in a slowly increasing mass of beta cells. The emphasis in this review is on recent insights on the natural history of the beta cell in a normal pancreas: sources of renewal, survival and changes in differentiation. PMID- 11042469 TI - Transplantation of the islets of Langerhans: new hope for treatment of type 1 diabetes mellitus. AB - For more than two decades, islet transplantation has been pursued as a curative treatment for type 1 diabetes mellitus (T1DM) with little success. It is likely that the failures of the past have involved technical difficulties in harvesting human islets, transplantation of insufficient amounts of islet tissue, the antagonistic effects of immune suppressive drugs, including calcineurin inhibitors and glucocorticoids, graft rejection and recurrent autoimmune disease. More recently, success has been reported in seven out of seven consecutive transplants using approaches that overcome the technical and therapeutic problems of the past. Although this success is noteworthy, issues remain that preclude the general application of islet transplants for treatment of the majority of patients with T1DM. These include the need for chronic immunosuppression and the requirement of large numbers of islets. Efforts are under way, using a variety of immunological, molecular and cellular strategies, to make this promising treatment available to the majority of patients with this disease. PMID- 11042470 TI - Searching for type 2 diabetes genes in the post-genome era. AB - The discovery of genes encoding maturity onset diabetes of the young (MODY) type 2 diabetes mellitus (T2DM) represents a milestone in the study of the genetics of complex diseases. The genes were revealed, in large part, through positional cloning and linkage analysis in families. The genes are relatively rare and have strong genotype-phenotype correlations. Based on the results of several genome scans for genes encoding T2DM in various racial-ethnic groups, it appears that, as in other complex diseases, multiple genes are involved, each contributing a small amount to the overall risk. New strategies for patient sampling, phenotyping, genotyping technologies and genetic analysis must be employed to further define the genetic basis of this disease. PMID- 11042471 TI - A cDNA RDA protocol using solid-phase technology suited for analysis in small tissue samples. AB - cDNA representational difference analysis (cDNA RDA) is a PCR-based subtractive enrichment procedure for the cloning of differentially expressed genes. In this study, we have further developed the procedure to take advantage of solid-phase technology, and to facilitate the use of RDA when starting material is limited. Several parameters of the PCR-based generation of cDNA representations were investigated, and a solid-phase based purification step was introduced to simplify removal of digested adapter-ends and uncleaved fragments. The use of magnetic particles increased the speed of the method, and also eliminated the risk of carry-over contamination between iterative steps of subtraction and PCR amplification. The modified protocol was evaluated in monitoring differences in gene expression in (i) a rat system consisting of livers with and without growth hormone treatment, and in (ii) a human system consisting of normal colon and colon cancer. PMID- 11042472 TI - A broad-host-range expression vector series including a Ptac test plasmid and its application in the expression of the dod gene of Serratia marcescens (coding for ribulose-5-phosphate 3-epimerase) in Pseudomonas stutzeri. AB - A series of expression vectors for gram-negative bacteria was constructed which combine broad-host-range, inducible expression from the tac promoter and diverse antibiotic resistance determinants. The tac promoter activity and the repression by lacIq can be quantitated with a separate test plasmid in the strain to be studied. The dod gene of Serratia marcescens was expressed in Pseudomonas stutzeri and was shown to code for D-ribulose-5-phosphate 3-epimerase. PMID- 11042473 TI - The application of direct immunofluorescence to intraoperative neurosurgical diagnosis. AB - A diagnostic problem can occur at the time of intraoperative consultation of neurosurgical tumors as to whether the tumor is of neuroectodermal origin or whether it represents an epithelial metastasis from another site. Intraoperative diagnoses based on hematoxylin and eosin stained frozen sections are often later confirmed by immunocytochemical analysis of formalin-fixed, paraffin-embedded tissue sections that are not available at the time of surgery. The objective of the current study was to demonstrate that the application of direct immunofluorescence to the intraoperative diagnosis of neurosurgical tumors would provide unequivocal, and nearly immediate results. This report describes a new application of an existing technique for an optimized, rapid procedure utilizing direct immunocytochemistry with fluorescence-labeled primary antibodies to analyze surgical biopsies intraoperatively. The examination of five neurosurgical biopsies established a neuroectodermal origin of three tumors via immunolabeling for glial fibrillary acidic protein (GFAP) and lack of labeling with keratin markers, whereas several metastatic lung carcinomas were identified by immunostaining for keratin, but not GFAP, markers. The results of the direct immunolabeling method were unequivocal and required only minutes. The same diagnoses were confirmed by standard immunocytochemical labeling of formalin fixed, paraffin-embedded sections, though it required several days to obtain the results. Direct immunofluorescence using fluorescently conjugated primary antibodies is a practical and rapid method for deciding whether a neurosurgical tumor is a primary glial or an epithelial metastatic tumor in origin. It is the first reported application of the technique for this aspect of rapid neurosurgical diagnosis. PMID- 11042474 TI - Micromachined interfaces: new approaches in cell immunoisolation and biomolecular separation. AB - As a novel therapeutic application of microfabrication technology, a micromachined membrane-based biocapsule is described for the transplantation of protein-secreting cells without the need for immunosuppression. This new approach to cell encapsulation is based on microfabrication technology whereby immunoisolation membranes are bulk and surface micromachined to present uniform and well-controlled pore sizes as small as 10 nm, tailored surface chemistries, and precise microarchitecture. Through its ability to achieve highly controlled microarchitectures on size scales relevant to living systems (from microm to nm), microfabrication technology offers unique opportunities to more precisely engineer biocapsules that allow free exchange of the nutrients, waste products, and secreted therapeutic proteins between the host (patient) and implanted cells, but exclude lymphocytes and antibodies that may attack foreign cells. Microfabricated inorganic encapsulation devices may provide biocompatibility, in vivo chemical and mechanical stability, tailored pore geometries, and superior immunoisolation for encapsulated cells over conventional encapsulation approaches. By using microfabrication techniques, structures can be fabricated with spatial features from the sub-micron range up to several millimeters. These multi-scale structures correspond well with hierarchical biological structures, from proteins and sub-cellular organelles to the tissue and organ levels. PMID- 11042475 TI - Surgical management of coarctation of the aorta in infants younger than five months: a study of fifty-one patients. AB - From January 1992 to December 1996, fifty-one (male 31) patients underwent operative intervention for coarctation of the aorta. The mean age was 26.8+/-20.3 days. Twenty six patients had simple coarctation and 25 patients had coexisting other complex cardiac anomalies. Coarctation with Hypoplastic aortic arch was seen in 47% of the patients. Resection and end-to-end anastomosis was done on 28 (55%), extended end-to end anastomosis on 22 (43.1%), and one had subclavian flap aortoplasty. The overall mortality was 13.7%. Five of the seven patients who died had additional congenital heart disease (p<0.05). The major cause of death was left ventricular dysfunction following surgery. The mean duration of follow-up was 16. 5+/-12.8 months. The rate of recoarctation was 19.6%. This study shows that associated cardiac anomalies increase the mortality. PMID- 11042476 TI - Do we need hypothermia in myocardial protection? AB - BACKGROUND: Since the concept of "elective cardiac arrest" has been introduced by Melrose et al., rapid arrest, hypothermia and additional protection has been employed in surgical myocardial protection in clinical and experimental settings. And cardioplegia technique employed these components improved clinical results of open heart surgery except special cases which require longer cardiac arrest. In 1991, Salemo et al. offered striking impact on most of cardiac surgeon with the report of retrograde continuous warm blood cardioplegia. Since then several reports pointed out the benefit of warm blood cardioplegia although inherent disadvantage of continuous cardioplegia were the inadequate visualization of the operative field. These reports recently lead some cardiac surgeon to intermittent warm blood cardioplegia. METHODS: This review introduced and examined our experimental and clinical data with reference to establish new surgical myocardial protection. CONCLUSIONS: Experimental and clinical data might encourage us to employ intermittent tepid (29 ) blood cardioplegia as a practical cardioplegia in open heart surgery. PMID- 11042477 TI - The role of cyclosporine A and interleukin-2 in obliterative airway disease in a rat tracheal transplant model. AB - The pathogenesis of obliterative bronchiolitis (OB) following lung and heart-lung transplantation remains unclear. We evaluated the role of CsA and IL-2 on the development of obliterative airway disease (OAD) by administrating exogenous IL-2 in a CsA-treated rat tracheal transplant model. Tracheal grafts were implanted into the peritoneal cavity from Brown Norway (BN) to BN rats or to Lewis (LEW) rats. Allotransplant: No treatment was given in group 1. Short-term CsA (25 mg/kg, i.m. on POD 2 and 3) was used in group 2. Group 3 was treated with long term CsA (25 mg/kg, i.m. on POD 2 and 3, followed by 5 mg/kg on POD 4 to 27). Administration of IL-2 (300, 000 IU/kg, i.p. on POD 15 to 19 and 22 to 26) was performed to long-term CsA treated rats in group 4. Isotransplant: No treatment was given to group 5, group 6 was treated with IL-2 (same regimen as in group 4). Grafts were harvested at different time points after Tx for histological assessment. No luminal obliteration was observed in group 5 and 6. Complete luminal obliteration was noted 4 weeks after Tx in group 1. In group 2 and 3, obliterative lesion occurred 4-6 weeks after CsA withdrawal. IL-2 increased epithelial loss, lymphocytic infiltration, and obliterative changes in group 4. Our results suggest that OAD is an immune mediated disorder. Furthermore, administration of exogenous IL-2 might be able to abrogate the protection from OAD by CsA therapy. PMID- 11042478 TI - Management of massive hemoptysis: a single institution experience. AB - BACKGROUND: Massive hemoptysis is a life threatening condition. Several therapeutic strategies have been applied in the clinical setting, with variable results. We reviewed our recent experience on this subject. MATERIAL AND METHODS: In a 5-year period, fifty-four patients (41 males, mean age 57.9 years) were treated for massive hemoptysis in our unit. The underlying pathology included bronchiectasis (n=31), active tuberculosis (n=9), pneumoconiosis (n=3), lung cancer (n=2) and pulmonary angiodysplasia (n=1). These patients often present with continuous bleeding with large volume of hemoptysis, or with recurrent episodes of bleeding. Bronchoscopic assessment and interventions were performed upon admission in all patients. Surgery was considered if the patient had acceptable pulmonary reserve and a bleeding source was clearly identified. If the patient was not considered fit for surgery, bronchial artery embolization was attempted. RESULTS: Hemoptysis ceased with conservative management in 7 patients (13%) only. Twenty seven (50%) patients received surgical resection. The procedures included lobectomy (n=21), bilobectomy (n=4) and pneumonectomy (n=2). The in-hospital mortality after surgery was 15%. Postoperative morbidity occurred in 8 patients, including prolonged ventilatory support, bronchopleural fistulae, empyema and myocardial infarction. Twenty-one patients not suitable for surgery were treated with bronchial artery embolisation, which was successful in 17 patients without any complications. CONCLUSION: The clinical outcome for massive hemoptysis reflects the generalized nature of a destructive disease process involving both lungs and a limited respiratory reserve. Surgery is associated with high risk of morbidity and mortality, and should be performed only in selected patients. Meanwhile, aggressive conservative therapy including bronchial artery embolization should be pursued. PMID- 11042479 TI - Effectiveness of prostaglandin E1 on pulmonary hypertension and right cardiac function induced by single-lung ventilation and hypoventilation. AB - It is known that prostaglandin E1 (PGE1) is a potent vasodilator and improves red cell deformability. Single lung-ventilation sometimes occurs under lung transplantation, lung cancer surgery and traumatic pneumonectomy, and may result in increased pulmonary resistance, right heart failure and severe hypoxemia. The present experimental study was undertaken to examine the effects of PGE1 on these states induced by single-lung ventilation and hypoventilation. Fourteen pigs weighing 32-33 kg were anesthetized, intubated and ventilated using a respirator and then randomly assigned to two groups, the control group and the PGE1 treated group, 7 pigs each. After median sternotomy to induce severe hypoxemia hypoventilation was induced and then the right hilus pulmonis was cross clamped. Mean blood pressure, mean pulmonary arterial pressure (PAP), pulmonary vessel resistance (PVR), right ventricular stroke work (RVSW) and arterial blood gases (PaO2 and SatO2) were measured at baseline, in the hypoventilation state, and 15 min, 1 hour, and 2 hours after the right hilus pulmonis clamping with hypoventilation. PGE1 (250 microg/20 ml saline) was administered via the central vein starting 15 min after right hilus cross clamping for 1 hour and 45 min in the PGE1 group. PGE1 significantly reduced PAP and PVR, normalized RVSW, and improved PaO2. PGE1 may be useful for the condition of increased pulmonary hypertension during single-lung ventilation and hypoventilation. PMID- 11042480 TI - Control system for an implantable rotary blood pump. AB - Rotary blood pumps can be used for long-term left ventricular assist devices. These pumps have several advantages over the conventional pulsatile pumps including smaller size, higher efficiency, and simple design and construction. However, one of the difficulties associated with the rotary blood pump is the proper control method to maintain an optimum flow rate in different physiological conditions. The rotary blood pump can be controlled by two methods. The first is to utilize the measured pump flow rate from its servo signal. The second is to detect and avoid abnormal pumping conditions such as; back flow and sudden increase in the pressure head. This abnormal situation typically occurs from excessive suction of blood when there is a functional or mechanical occlusion in the inflow cannula. The ultrasound flow meter is durable and reliable but it is difficult to continually monitor the blood flow rate of an implantable pump. Therefore, another method is needed instead of the continuous flow monitoring. One chronic calf having an LVAD was subjected for the development of this control system. This calf survived more than 6 months. Voltage, current, motor speed, heart rate and the pump flow rate were recorded and stored at 30-min intervals in a computer. Utilizing these parameters, attempts were made (1) to achieve indirect flow assessments and (2) to reveal abnormal operating parameters of the centrifugal pump (1). Indirect flow measurement, the predicted pump flow rate was calculated from these pump derived parameters (required power, motor speed and heart rate). The value of the coefficient of determination (R) between the measured and estimated pump flow rate was 0.796. (2) Abnormal operating indicator, there was an association between the required current and pump flow waves. The current was differentiated, and then calculated to the power of the differentiated current. The normal range of this value was 0.02+/-0.54. In abnormal conditions, this abnormal operating indicator increased 500 times. The predicted flow estimation method and abnormal operating indicator were available from intrinsic operating parameters of the pump and need no sensors. These two methods were simple, yet they are possibly effective and reliable servo control methods for a rotary blood pump. PMID- 11042481 TI - Clinical evaluation of ATS prosthetic valve by doppler echocardiography: comparison with St. Jude Medical (SJM) valve. AB - ATS valve is a bileaflet valve developed to have a superior function to traditional prosthetic valves on anti-thrombogenesis and hemolysis. The orifice area was enlarged in consequence of making its orifice with pyrolytic carbon materials, and the pivot was open pivot and the strut was eliminated. The efficiency of this valve has been reported in the Europe. In Japan, its clinical trial was performed in September 1993 and its clinical use was permitted in August 1996. In this study, we compared the functions of ATS and St. Jude Medical (SJM) valves with echocardiography. The pressure gradients at the mitral valve position were not significantly different between ATS and SJM valves, but that of ATS was lower than that of SJM. The pressure gradient at the aortic valve position of ATS was also lower than that of SJM, and especially those of the patients who performed aortic valve replacement with 23-mm ATS and SJM valves were 8.9+/-1.6 and 23.3+/-6.6 mmHg, showing a significant difference. Though further observation and review of this valve are needed at the chronic phase, ATS valve is an excellent bileaflet valve on the valvular function, showing a low pressure gradient. PMID- 11042482 TI - A case report of video-assisted thoracoscopic pneumonectomy for pulmonary atypical mycobacteriosis. AB - We report a case of a 61-year old woman with pulmonary atypical mycobacteriosis. Although she had been treated with chemotherapy, image findings gradually deteriorated. Chest X-ray and CT scan demonstrated that the advanced destruction with cavity formation and diffused shadow in the right lung. She underwent a right pneumonectomy under the video-assisted thoracic surgery (VATS) approach. The operation time was 4.9 hours, and the amount of intraoperative bleeding was 550 ml. The postoperative course was uneventful and the patient was discharged from hospital on 13th postoperative day. At present, two years after the operation, there is no progress of the pathogen into the other lung, and the patient is doing well with no postoperative complaint. Anatomically, pneumonectomy is a simple procedure, and in consideration of postoperative quality of life with alleviation of pain as well, VATS is worth trying in the cases where the status and nature of the disease are suitable for this technique. PMID- 11042483 TI - Extracardiac total cavopulmonary connection after superior cavopulmonary connection for left isomerism. AB - We report the case of a 3-year-old boy who underwent definitive conversion to the Fontan circulation after total cavopulmonary shunt using a total extracardiac right heart bypass. This simple, safe and reproducible procedure is an alternative to Fontan or total cavopulmonary connection procedure. PMID- 11042484 TI - A rare complication of PTCS: ruptured balloon with retained broken catheter. AB - This report describes a rare situation in which a retained ruptured balloon with broken catheter was caught in an incompletely opened stent during a percutaneous transluminal coronary stenting (PTCS) procedure in a 84 year old gentleman. A short, warm heart bypass and single cross clamping coronary artery bypass grafting procedure was performed. PMID- 11042485 TI - Off-pump coronary artery bypass grafting for the circumflex coronary artery via the left thoracotomy in redo CABG with the patent left internal thoracic artery graft to the left anterior descending artery. AB - Five patients had undergone off-pump coronary artery bypass grafting (CABG) as redo CABG via the left thoracotomy for the lesions of the left circumflex coronary arteries. In all patients, the internal thoracic artery (ITA) grafts to the LAD were well patent and acting significantly important in coronary circulation, however, ischemia due to the lesion of the LCX was significant. The saphenous vein grafts or the radial artery grafts were used as the materials of the grafts. The proximal ends of these grafts were anastomosed to the descending aorta. The procedures were completed successfully in all the patients and the excellent patency was shown angiographycally even in the long-term period after the surgery. Necessity of graft surgery only for the LCX lesion would be a rare occasion for a surgeon; however, these results suggest that the procedure is simple and less risky, which would encourage the surgeon to perform it in clinical situation. PMID- 11042486 TI - Ruptured sinus valsalva with infectious endocarditis: a technique of defect closure with an autologous-xenologous pericardial sandwich patch. AB - We report a case of ventricular septal defect with acquired rupture of sinus Valsalva induced by infectious endocarditis. After irrigati on of all infectious tissues, the defect was closed twice using two different patches. One was an autologous pericardial patch from the right ventricle and the other was a composite patch made of an autologous pericardium and axenologous pericardium from the left ventricle. As a result, the xenologous pericardium was sandwiched between autologous pericardiums. We thought that this "sandwiched patch" would compensate for the shortcomings of each type of pericardium and resist left ventricular pressure and infection. Despite the development of antibiotic therapy, infectious endocardit is (IE) is still one of the most difficult disease to cure. In the case of a rupture of sinus Valsalva, because of the rapid spread of infection into any of the cardiac chambers, surgical intervention is necessary. In this report, we describe a case treated successfully. PMID- 11042487 TI - Ruptured aortic dissecting aneurysm in Turner's syndrome: a case report and review of literature. AB - Cardiovascular malformations are frequently observed in Turner's syndrome. Bicuspid aortic valve and coarctation of the aorta are commonly associated with Turner's syndrome whereas aortic dissection is rare but its rupture results in death. We experienced a case of ruptured dissecting aneurysm (Stanford type A) in a 30-year-old female with Turner's syndrome. Emergent total arch replacement was performed successfully. A literature review revealed 32 cases of aortic dissection in patients with Turner's syndrome, including 15 cases of rupture. However, survival after rupture was reported only two cases. To our knowledge, this report descries the third known case of successful surgical management of ruptured aortic dissection in Turner's syndrome. PMID- 11042488 TI - Atherosclerotic aneurysm of the intrathoracic segment of the subclavian artery: a case report. AB - True aneurysms of the intrathoracic segment of the subclavian artery are extremely rare. Atherosclerosis is the most common etiology. The surgical approach and timing of repair remain controversial. We successfully treated a patient with a large proximal subclavian artery aneurysm which was secondary to atherosclerosis. The patient was asymptomatic for 30 years as the aneurysm enlarged. Three-dimensional computed tomography provided the most useful information regarding anatomy of the cervical vasculature. The patient underwent closure of the inlet port of the aneurysm and repair using a 6-mm Dacron bypass graft (Gelsoft , VASCTEK, Scotland) via a minimally invasive surgical approach. PMID- 11042489 TI - Fatty acids as uncouplers of oxidative phosphorylation. AB - This review summarizes data on the uncoupling effect of fatty acids on oxidative phosphorylation in mitochondria of various animal and plant tissues. PMID- 11042490 TI - A study of the Asp110-Glu112 region of EcoRII restriction endonuclease by site directed mutagenesis. AB - Site-directed mutagenesis of the ecoRII gene has been used to search for the active site of the EcoRII restriction endonuclease. Plasmids with point mutations in ecoRII gene resulting in substitutions of amino acid residues in the Asp110 Glu112 region of the EcoRII endonuclease (Asp110 --> Lys, Asn, Thr, Val, or Ile; Pro111 --> Arg, His, Ala, or Leu; Glu112 --> Lys, Gln, or Asp) have been constructed. When expressed in E. coli, all these plasmids displayed EcoRII endonuclease activity. We also constructed a plasmid containing a mutant ecoRII gene with deletion of the sequence coding the Gln109-Pro111 region of the protein. This mutant protein had no EcoRII endonuclease activity. The data suggest that Asp110, Pro111, and Glu112 residues do not participate in the formation of the EcoRII active site. However, this region seems to be relevant for the formation of the tertiary structure of the EcoRII endonuclease. PMID- 11042492 TI - Hypothetical 92.3-kD human protein homologous to angiotensin-converting enzyme: a new member of the zinc metalloprotease family? AB - Significant identity between a hypothetical 92.3-kD human protein and angiotensin converting enzyme (ACE; peptidyl-dipeptidase A; kininase II) has been found. Certain specific regions of the 92.3-kD protein indicate that this unidentified molecule may be a member of the zinc metalloprotease family. A method is suggested for determination of a structural and functional family of proteins with unknown structure and function. PMID- 11042491 TI - Expression, refolding, and ferritin-binding activity of the isolated VL-domain of monoclonal antibody F11. AB - Expression of the VL-domain of mouse monoclonal antibody F11 to human spleen ferritin in Escherichia coli cells is associated with the formation of insoluble protein aggregates (inclusion bodies). The aggregates were solubilized in the presence of guanidine hydrochloride and the recombinant VL-domain was purified by immobilized metal affinity chromatography (IMAC). Subsequent renaturation results in approximately 99% pure preparation with high yield. The VL-domain forms dimers at concentrations from 1 to 10 mg/ml. Monomeric form is detected only at protein concentrations below 0.5 mg/ml. Functional activity of the VL-domain was verified by two variants of ELISA. The affinity of the VL-domain ((0.2-1.2). 108 M(-1)) is similar to the affinity of the full-length parental antibody F11 because when the immobilized VL-domain was used, the binding constant of ferritin to the VL-domain was only 4-6-fold lower than that in the case of F11 antibody. In another ELISA system with immobilized ferritin, affinity was decreased 30-fold. The VL-domain of antibody F11 is the first example of the recombinant variable domain of the immunoglobulin light chain that preserves the antigen-binding activity in the absence of the partner VH-domain. The data indicate that the recombinant VL domain can be used in construction of chimeric immunotoxins and other antigen binding proteins in immunotherapy and in studies of correlations between folding, stability, and activity of immunoglobulins. PMID- 11042493 TI - Regulation of oxidative phosphorylation in the inner membrane of rat liver mitochondria by calcium ions. AB - The effect of accumulation of Ca2+ at physiological concentrations (10(-8)-10(-6) M) on the rates of ATP synthesis and hydrolysis in rat liver mitochondria was studied. An addition of 5 x 10(-7) M Ca2+ resulted in the maximal rates of synthesis and hydrolysis of ATP. Decrease in the concentration of Ca2+ to 10-8 M or its increase to 5 x 10(-6) M inhibited oxidative phosphorylation and ATP hydrolysis. It was found that the rate of oxidative phosphorylation correlated with the phosphorylation level of a 3.5-kD peptide in the mitochondrial inner membrane on varying the Ca2+ concentration. The possible regulation of oxidative phosphorylation in mitochondria by Ca2+ is discussed. PMID- 11042494 TI - Effects of nitric oxide donors on Ca2+-dependent [14C]GABA release from brain synaptosomes: the role of SH-groups. AB - Nitric oxide (NO) modulates processes of synaptic transmission at pre- and postsynaptic levels. In the present work we studied the mechanisms of action of NO on [gamma-14C]amino-n-butyric acid ([14C]GABA) release in rat cortical synaptosomes. NO donors--S-nitroso-L-cysteine and hydroxylamine (but not sodium nitroprusside)--inhibited the neurotransmitter efflux in a concentration range from 10 microM to 1 mM. Nitrosocysteine completely and selectively suppressed the Ca2+-dependent (vesicular) [14C]GABA release, while not affecting the Ca2+ independent component of the [14C]GABA transport. The influence of NO donors was not related to activation of guanylyl cyclase, since the membrane-permeable cGMP analog dibutyryl-cGMP did not mimic and the guanylyl cyclase inhibitor methylene blue did not change the NO effects. In contrast, the membrane-permeable SH reagent N-ethylmaleimide (NEM) resembled the effects of NO donors on the Ca2+ dependent [14C]GABA release. The degree of inhibition of the release by nitrosocysteine, hydroxylamine, and NEM correlated with their ability to oxidize intra-synaptosomal SH-groups. These data suggest that synaptosomal sulfhydryl groups are the target for NO action at the presynaptic level. The NO-induced oxidation of thiols may be involved in physiological and, especially, pathological effects of nitric oxide in the central nervous system. PMID- 11042495 TI - Intracellular glucosaminidase of the bacterium Xanthomonas campestris IBPM B-124: purification and properties. AB - A system of intracellular autolytic enzymes of the bacterium Xanthomonas campestris IBPM B-124 was found to include enzymes with muramidase and glucosaminidase activities, while a system of extracellular bacteriolytic enzymes of the same bacterium includes muramidase, muramoylalanine amidase, and endopeptidase. Using a purification technique including fractional precipitation with ammonium sulfate, gel-filtration on Toyopearl HW-55F, and FPLC ion-exchange chromatography on Mono Q, a preparation of intracellular glucosaminidase was purified 435-fold with 16% yield (SDS-PAGE data indicated the presence of minor protein contaminants). Some physicochemical properties of the purified enzyme were determined: molecular mass 26 kD, Km = 5.6 x 10(-4) M with p-nitrophenyl-2 acetamido-2-deoxy-beta-D-glucopyranoside as the substrate, and pH optimum 8.0 8.5. The enzyme is active over a wide range of Tris-HCl buffer concentrations (0.01-0.5 M) and has temperature optimum at 37-40 degrees C. The glucosaminidase activity is sensitive to p-chloromercuribenzoate (PCMB), phenylmethylsulfonyl fluoride (PMSF), and the disodium salt of ethylenediamine tetraacetic acid (EDTA). The properties of this glucosaminidase markedly differ from those of all extracellular bacteriolytic enzymes of Xanthomonas campestris. These findings indicate that the system of autolytic enzymes of this bacterium functions independently and is not connected with the system of extracellular bacteriolytic enzymes. PMID- 11042496 TI - Conformational analysis of the biologically active RGD-containing anti-adhesive peptide cyclo(ArgGlyAspPhe-D-val). AB - Using theoretical conformational analysis, the RGD-peptide with anti-adhesive activity cyclo(ArgGlyAspPhe-D-Val) was studied. Random sampling was used to search the conformational space of the allowed torsional angles of the main chain of the molecule. Among 900 stable conformers with different folding of the cyclic moiety of the peptide, only those were selected which corresponded to low-energy conformers of the model linear tripeptide AcAlaGlyAspNHMe. This peptide served as the main chain template of the RGD-fragment of the studied cyclopeptide molecule. Of 36 selected cyclopeptide conformers with potential biological activity, only a few contain stable intramolecular hydrogen bonds. It was supposed that a biologically active conformer of the cyclopeptide molecule exists in solution among other conformers, but not necessarily as the major component of the equilibrium mixture. PMID- 11042497 TI - Interaction of trypsin with multilamellar vesicles of soybean lipids. AB - The pH dependence of complex formation of trypsin with multilamellar vesicles (MLV) of soybean lipids has been investigated. The lipids were characterized by the same phospholipid composition, but the content of other lipids differed. Decrease of pH or introduction of negatively charged components into the lipid samples increased trypsin content in the protein-lipid complexes. This suggests electrostatic interaction between the protein and soybean lipids. The dependence of trypsin activity in the complexes with MLV on their concentration and on the presence of an ionic detergent was studied. Trypsin-MLV interaction did not result in complete inactivation of the protein molecules. Moreover, the effects of dilution and addition of ionic detergent on trypsin activity were additive. Using a fluorescence technique, complex formation with MLV was found to stabilize trypsin molecules, preventing their autolysis. PMID- 11042498 TI - Structure of the O-specific polysaccharide of the bacterium Proteus vulgaris O23. AB - An acidic O-specific polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of the bacterium Proteus vulgaris O23 (strain PrK 44/57) and found to contain 2-acetamido-2-deoxy-D-galactose, 2-acetamido-2-deoxy-D-glucose, and D-galacturonic acid. Based on 1H- and 13C-NMR spectroscopic studies, including two-dimensional correlation spectroscopy (COSY), total correlation spectroscopy (TOCSY), nuclear Overhauser effect spectroscopy (NOESY), and 1H,13C heteronuclear multiple-quantum coherence (HMQC) experiments, the following structure of the branched tetrasaccharide repeating unit of the polysaccharide was established: [figure], where the degree of O-acetylation of the terminal GalA residue at position 4 is about 80%. A structural similarity of the O-specific polysaccharides of P. vulgaris O23 and P. mirabilis O23 is discussed. PMID- 11042499 TI - Structure of an acidic O-specific polysaccharide of the marine bacterium Pseudoalteromonas sp. KMM 634. AB - An acidic O-specific polysaccharide containing D-glucuronic acid (D-GlcA), 2,3 diacetamido-2,3-dideoxy-D-glucuronic acid (D-GlcNAc3NAcA), 2,3-diacetamido-2,3 dideoxy-D-mannuronoyl-L-alanine (D-ManNAc3NAcA6Ala), and 2-acetamido-2,4, 6 trideoxy-4-[(S)-3-hydroxybutyramido]-D-glucose (D-QuiNAc4NAcyl) was obtained by mild acid degradation of the lipopolysaccharide of the bacterium Pseudoalteromonas sp. KMM 634 followed by gel-permeation chromatography. The polysaccharide was cleaved selectively with a new solvolytic agent, trifluoromethanesulfonic acid, to give a disaccharide and a trisaccharide with D GlcNAc3NAcA at the reducing end. The borohydride-reduced oligosaccharides and the initial polysaccharide were studied by GLC-MS and 1H- and 13C-NMR spectroscopy, and the following structure of the linear tetrasaccharide repeating unit of the polysaccharide was established: -->3)-alpha-D-QuipNAc4Ac4NAcyl-(1-->4)-beta-D ManpNAc3NAcA6Ala+ ++-(1-->4)-b eta-D-GlcpNAc3NAc3NAcA-(1-->4)-beta-D-GlcpA-(1-->. PMID- 11042500 TI - Structure and folding of bacteriophage T4 gene product 9 triggering infection. II. Study Of conformational changes of gene product 9 mutants using monoclonal antibodies. AB - Gene product 9 (gp9) of bacteriophage T4, whose spatial structure we have recently solved to 2.3 A resolution, is a convenient model for studying the folding and oligomerization mechanisms of complex proteins. The gp9 polypeptide chain consists of 288 amino acids forming three domains. Three monomers, packed in parallel, assemble to a functionally active protein. The main aim of this work was to study conformational changes and trimerization of gp9 deletion mutants using monoclonal antibodies (mAbs). We selected a set of mAbs interacting with the amino, middle, and carboxyl regions of the protein, respectively. Eighteen mAbs bind to native as well as to denatured protein, and two mAbs bind to denatured protein only. Using mAbs, we found that deletions of the gp9 N-terminal region result in conformational changes in the middle and C-terminal domains. The study of mAb binding to the CDelta. truncated mutant by competitive ELISA and immunoblotting shows that the C-terminus of the gp9 sequence is essential for protein trimerization and stability. A single point substitution of the Gln282 residue causes formation of a labile trimer that has significant conformational changes in the protein domains. The results of our study show that folding and trimerization of gp9 is a cooperative process that involves all domains of the protein. PMID- 11042501 TI - The primary structure of the N-terminal region of mature alkaline phosphatase is critical for secretion and function of the enzyme. AB - The export signal has been assumed to be localized not only in the signal peptide of a secreted protein precursor, but also in the N-terminal region of the mature polypeptide chain. Mutant alkaline phosphatases with amino acid substitutions of two positively charged residues (Lys or Arg) in this region at different distances from the signal peptide have been studied to test this assumption. The efficiency of secretion has been shown to decrease in mutant proteins with amino acid substitutions in the region of 16-18 amino acid residues; the closer to the signal peptide is the substitution, the greater is the decrease. A change in the primary structure of the N-terminal domain results also in an increase in the Michaelis constant, which is greater the farther is the amino acid substitution from the signal peptide, suggesting a change in the enzyme function as well. PMID- 11042503 TI - Monoamine oxidase inhibition by N-alkyloxycarbonyl derivatives of ethylene diamine. AB - Urethane type derivatives of ethylene diamine (EDA) were synthesized and tested as inhibitors of rat liver mitochondrial monoamine oxidase (MAO) A and B. The nature of the aromatic ring and the position of substituents in it were crucial for manifestation of the inhibitory activity. 3,4- and 2,4 Chlorobenzyloxycarbonyl-EDA derivatives were the most potent MAO A inhibitors. The inhibition of both MAO A and to a lesser extent MAO B depended on preincubation time with these inhibitors. The activity of both enzymes did not recover completely after repeated sedimentation and resuspension of inhibitor treated mitochondria. The data suggest that these compounds exhibit properties of tight-binding reversible inhibitors of MAO A and B. The development of a new generation of MAO inhibitors causing simultaneous reversible nonselective inhibition of MAO A and B must meet one important criterion, the same type of inhibition of both the enzymes. PMID- 11042502 TI - Interaction of bacterial endotoxins with chitosan. Effect of endotoxin structure, chitosan molecular mass, and ionic strength of the solution on the formation of the complex. AB - The interaction of endotoxins of different structure (lipopolysaccharides (LPS) and lipopolysaccharide-protein complexes (LPPC)) with chitosan has been studied. It was shown that the mechanism of interaction is rather complicated and depends on the macromolecular organization of endotoxin as well as on the degree of polymerization of the chitosan. Chitosan with molecular mass of 20 kD reveals higher affinity to LPS than chitosan with molecular mass of 140 kD. Endotoxins with long O-specific chains can bind completely with chitosan with the formation of LPS-chitosan and LPPC-chitosan complexes with weight ratios between the original components of 1:1 and 1:5. When endotoxins with higher degree of hydrophobicity and short O-specific chains were mixed with chitosan, a part of the LPS remained unbound. The stability of the complexes formed depends on ionic strength. It was shown that, in addition to electrostatic forces, other types of forces take part in the formation of the complexes. A decrease in acute toxicity of various LPSs is observed on their binding with chitosans. PMID- 11042504 TI - The major phospholipid of Escherichia coli, phosphatidylethanolamine, is required for efficient production and secretion of alkaline phosphatase. AB - The major phospholipid of the Escherichia coli membranes--the zwitterion phosphatidylethanolamine (PE)--is the only phospholipid involved in the formation of non-bilayer structure of membrane lipids, which is supposed to be necessary for efficient translocation of secreted proteins across the cytoplasmic membrane. The effect of PE on the production and secretion of alkaline phosphatase has been studied in this work using the mutant strain E. coli AD93, which is unable to synthesize PE. It was shown that this phospholipid is required for the efficient production and secretion of alkaline phosphatase. The anionic phospholipid cardiolipin in combination with divalent cations Mg2+ functionally replaces PE in these processes, participating in the regulation of lipid polymorphism. PMID- 11042505 TI - Kinetics of inhibition of green crab (Scylla serrata) alkaline phosphatase by vanadate. AB - Green crab (Scylla serrata) alkaline phosphatase is a metalloenzyme that catalyzes the nonspecific hydrolysis of phosphate monoesters. The kinetics of inhibition of the enzyme by vanadate has been studied. The time course of the hydrolysis of p-nitrophenyl phosphate catalyzed by the enzyme in the presence of different Na3VO4 concentrations showed that, at each Na3VO4 concentration, the rate decreased with increasing time until a straight line was approached, the slopes of the straight lines being the same for all concentrations. The results suggest that the inhibition of the enzyme by Na3VO4 is a slow, reversible reaction with fractional residual activity. The microscopic rate constants were determined for the reaction of the inhibitor with the enzyme. As compared with Na2HPO4 (Ki = 0.95 mM), Na2HAsO4 (Ki = 1.10 mM), and Na2WO4 (Ki = 1.55 mM), the results suggest that Na3VO4 (Ki = 0.135 mM) is a considerably more potent inhibitor than other inhibitors. PMID- 11042506 TI - Expression of Cell Cycle Regulating Proteins in an Unusual Transformation of Mantle Cell Lymphoma. AB - We describe here two patients with mantle cell lymphoma (MCL) who after a few years, developed to the diffuse large cell lymphoma (DLCL)( anaplastic centrocytic lymphoma) growing in a diffuse sheets without the classical MCL component. In both the initial and second biopsy specimens, in each case, tumor cells were positive for cyclin D1, sIgM, sIgD, and CD5, but were negative for CD10 and CD23. In a study of immunoglobulin heavy chain (IgH) gene rearrangement, using the polymerase chain reaction (PCR) method, the products obtained from each paired biopsy tissue sample were the same size, and in one case had an identical sequence to the non-mutated VH gene. Immunohistochemistry was used to examine the expression of p53, p27(Kip1) and cyclin E. Interestingly, there was clear overexpression of p53 protein in case 1 but not in case 2, compared with other typical MCL cases. The expression of p27(Kip1) in the second biopsies of each case was decreased compared with those in the initial biopsies. In case 2, however, p27(Kip1) was clearly expressed in the first and second biopsies, in contrast to other typical MCL cases. Thus these 2 cases demonstrate not only that the variant form of MCL may arise de novo, but also that MCL may transform to DLCL at the time of relapse. Although the mechanism of tumor progression/transformation is still poorly understood, the overexpression of p53 or p27(Kip1) may be linked to a cellular mechanism involved in the development of the variant form of MCL. PMID- 11042507 TI - CD40 ligand in CLL pathogenesis and therapy. AB - Advances in immunology during the past three decades have facilitated our understanding of the biology of specific lymphoid neoplasms including chronic lymphocytic leukemia (CLL). Investigations in our laboratory have focused on CD40, a critical regulator of B cell survival and differentiation, and its ligand, CD154 (CD40L). We have established that in some cases of CLL the malignant cells express both CD40 and CD154, and on the basis of those observations, proposed a model for CLL tumor growth due to CD40-CD154 interactions within and among the malignant cells, and for the occurrence of autoimmune syndromes in some cases of CLL. Here, we include an update on our studies regarding CD154 expression in CLL, a review of the data regarding the consequences of CD40 engagement in CLL B cells, and a discussion of these findings in the context of the complex and potentially opposite outcomes that have been reported for CD40-mediated signals in CLL. The implications for therapy, such as by impedance to CD154-CD40 interaction using antibody to CD154, or by selective inhibitors of NF-kappa B, are considered. PMID- 11042508 TI - Clinical and biologic diversity of leukemias occurring in patients with mastocytosis. AB - Patients with systemic mast cell (MC) disease, but not those with cutaneous mastocytosis, are at a high risk (10-30%) to develop life-threatening myelogenous malignancies. In a significant proportion of cases, myeloid leukemias occur. Using conventional criteria, such leukemias resemble acute myeloid leukemia (AML), chronic myeloid leukemia (CML), or myelomonocytic leukemia (CMML). Mast cell leukemia (MCL) may also occur. Myeloid leukemias (AML, CML, CMML) can develop in indolent or aggressive mastocytosis (skin lesions present or absent) with a variable prephase of MC disease. By contrast, MCL (typically without skin lesions) often develops on a "de novo" basis, and, if at all recognized, a prephase resembling (malignant) mastocytosis, is short. MCL differs from myeloid leukemias (AML, CML, CMML) by morphologic and phenotypic cellular characteristics. In fact, MCL are strongly tryptase-positive, c-kit-positive, myeloperoxidase (MPO) -negative neoplasms with variable metachromasia and chloroacetate esterase expression, whereas an MPO-positive, tryptase-negative phenotype supports the diagnosis of a myeloid non-MC lineage disease. Thus, MCL, but also myeloid non-MC lineage leukemias can develop in patients with (systemic) mastocytosis. Little is known, however, about the pathophysiologic basis of co evolution. In the present article, the concomitant occurrence of mastocytosis and leukemia is discussed in the light of the literature and of concepts proposed to explain the biologic basis of this phenomenon. PMID- 11042509 TI - Partially matched transplants with allogeneic CD34+ blood cells. AB - Allogeneic CD34(+) selected cell transplantation from a partially matched donor is feasible for the treatment of hematopoietic malignancies. Severe graft-versus host disease (GvHD) can be prevented by this approach, and a lengthy search for an unrelated donor can be avoided. However, long-term engraftment is still a challenge, and may require intensified conditioning regimens with severe toxicities to sustain engraftment. Furthermore, unacceptable effects such as relapse and life-threatening viral infections might be more frequent after such transplants compared with non-T-cell-depleted transplants from an HLA-identical donor. Strict indications should be considered for this treatment based on a risk/benefit assessment for very high-risk patients. The early application of periodic donor leukocyte infusion (DLI) may be useful for preventing both late graft rejection and serious viral infections. PMID- 11042510 TI - Pathogenesis of HIV-1 infection within bone marrow cells. AB - Mononuclear phagocytic cells and CD4+ T lymphocytes represent the major targets for infection by HIV-1 in vivo. The most severe pathogenic features associated with HIV-1 infection can be attributed to malfunction or premature death of these cells that are of hematopoietic origin. Patients with acquired immunodeficiency syndrome (AIDS), suffer from many hematologic disorders, particularly those persons with long-term infection of HIV-1. These disorders include anemia, lymphocytopenia, thrombocytopenia and neutropenia. The mechanisms that lead to the induction of these disorders are multi-factorial. However, sufficient evidence has accumulated which suggests that HIV-1 infection of cells within the microenvironment of the bone marrow can lead to the induction of hematopoietic deficits. Most studies indicate that marrow-derived hematopoietic stem cells cannot be infected by HIV-1 until they undergo modest differentiation in order to express the appropriate receptors to enable virus entry and subsequent replication. Some cells within the mixed environment of the marrow stroma appear to support HIV-1 replication however. These cells include marrow microvascular endothelial cells, sometimes referred to as blanket cells, stromal fibroblasts, as well as mononuclear phagocytes. Our recent experiments suggest that the HIV-1 accessory protein, Vpr, plays some role in the activation of marrow-derived mononuclear phagocytes which appears to result in premature phagocytosis of non adherent marrow cells present in the in vitro cultures. This phenomenon could account, in part, for the induction of cytopenias that are typical of individuals infected by HIV-1. PMID- 11042511 TI - Structural and functional studies on the leukemia inhibitory factor receptor (LIF R): gene and soluble form of LIF-R, and cytoplasmic domain of LIF-R required for differentiation and growth arrest of myeloid leukemic cells. AB - The leukemia inhibitory factor receptor (LIF-R) subunit is a component of cell surface receptor complexes for the multifunctional cytokines, LIF, cardiotrophin 1, ciliary neurotrophic factor, and human oncostatin M. The structure of the human LIF-R gene is similar to that of the mouse gene. The transmembrane receptor is encoded by 19 exons. Two distinct 5' non-coding exons are present, indicating the existence of alternative promoters. An extra-exon specific to the mouse soluble receptor contains a stop codon and polyadenylation signals in a B2 repetitive element. On the other hand, LIF-R mRNAs containing unspliced introns are abundantly present in human tissues. These intronic sequences introduce a termination codon before the transmembrane domain. Human choriocarcinoma cells expressing these mRNAs release soluble LIF-R. The cytoplasmic domain of LIF-R can generate the signals for growth arrest and differentiation of mouse myeloid leukemic cells when they are induced to form a homodimer of the cytoplasmic domain independently of gp130. Two membrane-distal tyrosines on the YXXQ motif of LIF-R are critical not only for STAT3 activation but also for growth arrest and macrophage differentiation of WEHI-3B D+ cells. PMID- 11042512 TI - Leukemia relapse reconsidered from the molecular aspect. AB - Relapse, a major obstacle in the treatment of acute leukemia, is essentially caused by re-growth of residual leukemia cells, frequently accompanied by resistance to chemotherapy. Comparative studies of clones both at initial diagnosis and at subsequent relapse have indicated that phenotype and karyotype are frequently changed at relapse. This can be recognized as the result of negative selection by chemotherapy in a heterogeneous population. Furthermore, complex molecular alterations that include the loss of as well as the acquisition of mutations are noticed by comparing multiple genes associated with leukemia, suggesting a continuous genetic evolution. Studies on leukemia relapse have thus served as a model of clonal progression, which can be serially observed, including selection by chemotherapy, induction of resistant phenotype, and genetic alteration. PMID- 11042513 TI - Immune reconstitution following allogeneic peripheral blood progenitor cell transplantation. AB - Delayed immune reconstitution following allogeneic stem cell transplantation remains a major clinical problem, resulting in significant transplant-related mortality from infectious complications. The recovery of immunity after bone marrow transplantation (BMT) is a complex process dependent on a large number of pre- and post-transplant factors. It has been suggested that the use of peripheral blood instead of bone marrow as stem cell source may accelerate immune reconstitution after allogeneic transplantation. Some authors have recently reported a more rapid recovery of the number and function of T and B cells after allogeneic peripheral blood progenitor cell transplant (allo-PBPCT) in comparison with conventional BMT, results which would reflect the high number of lymphocytes infused to the patients. Such a rapid immune recovery could indeed contribute to the apparent therapeutic advantage of PBPCT when compared with BMT. However, there is limited knowledge on this issue and randomized trials are required to prove whether allo-PBPCT is indeed superior to BMT in terms of immune reconstitution post-transplant. A review of some quantitative and functional aspects of immune recovery after allo-PBPCT is presented in this article. PMID- 11042514 TI - Chemotherapy of secondary leukemias. AB - Chemotherapy of secondary leukemias is currently still considered to be associated with poor results. However, recent data suggest that the response to remission induction may substantially differ according to the previous medical history of the patients. Therapy related leukemia, arising following exposure to previous alkylating agents or radiotherapy, is often associated with chromosomal abnormalities involving chromosomes 5 and 7 and has a particularly bad response, whereas AML after exposure to epipodophyllotoxins or topoisomerase-II active agents could have a somewhat better response. Acute promyelocytic leukemia secondary to treatment of a primary malignant neoplasm seems to be associated with a better response if compared to other cytotypes of AML or to AML arising after transformation of myelodysplasia. However, here the literature data are not in full agreement, as different kinds of approaches have been applied. In fact, even if the problems encountered in treating patients with secondary leukemia are similar to those seen in patients with AML arising in a background of myelodysplasia (resistant disease and prolonged cytopenia after treatment), there are data suggesting that the use of high dose ara-C, with or without fludarabine, can circumvent resistance in a small but significant number of cases. One of the unsolved problems which still remains is how to consolidate the CR induced with high dose ara-C or with cycles based on anthracycline derivatives. In addition, another question relates to the categories of patients in whom chemotherapy may change the expected survival. Intensive post-remission chemotherapy, with or without autologous HSCT, may constitute an appropriate alternative for patients lacking a suitable sibling donor or for older patients who are in remission after chemotherapy and also able to tolerate other cycles of intensive chemotherapy. In this respect, the specific cytogenetic abnormality involved should be considered the most important prognostic factor for response and disease free survival; patients with abnormalities of chromosome 5 and 7 have a particularly low possibility of response and duration of CR. Furthermore, it is still debatable whether patients, especially the elderly, with these characteristics should go through a series of conventional treatments or just receive supportive treatment. On the other hand, patients with better prognostic factors should be entitled to further intensive treatments, taking into account possible delayed recovery and/or possible less successful collection of peripheral or marrow stem cells. PMID- 11042515 TI - Impact of chemotherapy on the mobilisation, harvest and economic costs of autologous peripheral stem cell transplantation in patients with multiple myeloma. AB - To evaluate factors affecting mobilisation and harvest and to calculate the economic costs of autologous stem cell transplantation in multiple myeloma (MM) we analysed 29 consecutive patients who had been transplanted at the Nijmegen University Hospital between January 1992 and February 1999. Thirteen patients had been treated with three or more melphalan cycles before transplantation (melphalan group), while four of the remaining 16 patients (no-melphalan group) had only received one melphalan cycle with an interval of one year or longer before harvest. The two groups were analysed for differences in mobilisation, harvest and the costs. Collection of a sufficient number of peripheral stem cells failed in 4 patients in the melphalan group, and these patients were transplanted with both bone marrow and peripheral stem cells. The greater need for growth factors (median 6,400 microg vs 4,500 microg) and the longer duration of admission (median 8 days vs 3 days) for mobilisation in the melphalan group increased significantly (p=0.01) the total mobilisation costs (median fl 13,876 vs fl 6,101). The greater number of apheresis sessions (median three) and the additional bone marrow harvests for patients who could not achieve a sufficient number of stem cells, increased significantly (p<0.001) the total harvest costs (median fl 9,690 vs fl 1,615) in the melphalan group. This resulted in significantly (p=0.008) higher overall costs of the procedure (median: fl 49,576 vs fl 35,889). The haematopoietic recovery of all groups was similar. The no melphalan group was subdivided in two groups based on the median number of chemotherapy cycles before harvest. The heavily treated group had received more than 5 chemotherapy cycles and the moderately treated group four cycles or less. The median overall costs of these two groups were comparable (median fl 36,837 vs fl 34,351). This study suggests that the administration of stem cell toxic melphalan before harvest contributes to administration of more dosages of growth factor, longer admission duration for mobilisation and higher number of leukaphereses in order to collect sufficient number of stem cells. This resulted in significantly higher overall costs. More cycles of chemotherapy without melphalan did not increase significantly any studied parameter nor the total costs of procedure. Melphalan therapy or heavy pre-treatment did not prolong the repopulation interval, probably due to the infusion of similar number of progenitor cells. PMID- 11042516 TI - High dose ifosfamide in combination with etoposide and epirubicin followed by autologous stem cell transplantation in the treatment of relapsed/refractory Hodgkin's disease: a report on toxicity and efficacy. AB - Patients with Hodgkin's disease (HD) refractory to first line chemotherapy and those who have rapid or multiple relapses have a very poor prognosis. With the increasing use of hybrid chemotherapy these patients will have been exposed to many of the drugs active in HD so it is important to develop salvage regimens that are novel and demonstrate activity in this group of patients. We report the use of a continuous high dose infusion of ifosfamide at a dose of 9g/m(2) over 3 days in combination with etoposide and epirubicin followed by autologous stem cell transplant with either BEAM or Melphalan/VP16 conditioning in this difficult group. Forty six patients (28M:18F) with a median age of 28 years (range 13-45) were treated. Overall 39 out of 46 (85%) patients responded to treatment, with 17 achieving complete remission and 11 a good partial remission; 28 proceeded to autologous bone marrow/stem cell transplantation. In total, 23 patients are alive and in continuous remission with a follow up of between 12 and 61 months. Median overall survival for the whole group is 36 months. Haematological toxicity, particularly neutropenia (WHO grade IV), was observed in all cases but improved over the 3 courses of treatment in all patients. Non-haematological toxicity was not a major problem; no significant cardiac, hepatic, renal, pulmonary or neuro toxicity was observed and there were no deaths on treatment. This regime shows promise in patients with difficult Hodgkin's disease and warrants further study. PMID- 11042517 TI - Atypical lymphocytes in B-cell chronic lymphocytic leukemia and trisomy 12 studied by conventional staining combined with fluorescence in situ hybridization. AB - Trisomy 12 is one of the most frequent chromosomal abnormalities in B-cell chronic lymphocytic leukemia (CLL), and is predominantly found in CLL with atypical morphology (aCLL). It has been suggested that the atypical morphology might be a feature of the abnormal trisomy 12 clone, but so far it has been difficult to allocate chromosomal aberrations to individual leukemic cells identified by cytomorphology. We therefore wanted to use our MGG/FISH method, which combines fluorescence in situ hybridization (FISH) and standard cytomorphology, to study if the trisomy 12 clone in CLL was restricted to lymphocytes with atypical morphology. Peripheral blood specimens of four patients with aCLL were studied using a DNA probe against the pericentromeric region of chromosome 12. Trisomy 12 was found in 10-34 % of the lymphocytes. In three patients, the proportion of atypical and typical lymphocytes with trisomy 12 was quite comparable, and so was the percentage of atypical cells with lymphoplasmacytoid morphology and those with cleaved nucleus showing trisomy 12. Only one patient differed, since we found an overrepresentation of trisomy 12 among the atypical lymphocytes. However, this could be fully explained by the diluting effect of contaminating T-cells after chemotherapy. The results of the present study show that despite the strong association of trisomy 12 and atypical morphology in CLL, this chromosomal abnormality is not confined to lymphocytes with atypical morphology, but is also found in typical CLL cells. This supports that both cell types have the same clonal origin and that different cell morphology cannot be explained alone by the acquisition of an additional chromosome 12. PMID- 11042518 TI - Outcome of clonal hemophagocytic lymphohistiocytosis: analysis of 32 cases. AB - We studied the impact of clonality, determined by analysis of Epstein-Barr virus genome termini, T-cell receptor genes and clonal chromosomal abnormality, on the clinical outcome in 32 patients with hemophagocytic lymphohistiocytosis (HLH). Of the cases studied, 23 cases were EBV-clonal, 15 cases were TCR-clonal and 7 cases were cytogenetically clonal. Thirty patients were treated with immuno chemotherapy and/or multiagents' chemotherapy and 4 received bone marrow transplantation. All 7 cases, in which cytogenetically abnormal clones were identified, were fatal (3-year survival by Kaplan-Meier analysis; 14%, 95%CI: 0 40%). None of these 7 cases received bone marrow transplantation. On the other hand, the 3-year survival of 23 clonal EBV-positive HLH cases including 4 cytogenetically abnormal cases was 64 % (95%CI: 42-84%), while that of 15 TCR clonal cases was 53% (95%CI: 26-78%). Our observations suggest that cytogenetically abnormal cases are at extremely high risk, requiring intensive immuno-chemotherapy followed by prompt and timely allogeneic bone marrow transplantation. PMID- 11042519 TI - Serum CD44 levels preceding the diagnosis of non-Hodgkin's lymphoma. AB - Serum CD44 (s-CD44) concentrations were measured in sera taken from 49 individuals who were diagnosed with non-Hodgkin's lymphoma 0.9 to 7.2 years after taking the blood sample, and from 49 controls matched for age. The serum samples had been collected in conjunction of the Alpha-Tocopherol, Beta-Carotene Cancer Prevention (ATBC) study, which evaluated the influence of vitamin supplementation on cancer prevention. S-CD44 was measured using chemiluminescence enzyme immunoassay. S-CD44 concentrations of the cases were significantly elevated before the diagnosis of lymphoma when compared to the serum levels found in the controls (median, 447 ng/mL; range, 108-780 ng/mL vs. median, 364 ng/mL; range, 53-660 ng/mL; p=0.012). Individuals who were later diagnosed with high grade lymphoma according to the Kiel classification (n=21) had significantly higher values than the controls 0.9-4.0 years before the diagnosis, but such a difference could not be detected if serum samples had been taken more than 4 years before the diagnosis. The s-CD44 levels were not significantly elevated among individuals who were later diagnosed with low grade malignant non-Hodgkin's lymphoma (n=25) as compared to their controls. The prediagnostic s-CD44 levels in cases and controls overlapped markedly, and a value higher than the highest value found among the controls (660 ng/mL) was found only in 5 (10%) samples taken from individuals who were later diagnosed with lymphoma. We conclude that serum CD44 may be elevated a few years preceding the diagnosis of non-Hodgkin's lymphoma, but the levels overlap markedly with those found in individuals without lymphoma. PMID- 11042521 TI - Diffuse alveolar hemorrhage in acute promyelocytic leukemia patients treated with ATRA--a manifestation of the basic disease or the treatment. AB - All-trans-retinoic acid (ATRA) is considered the recommended induction treatment for acute promyelocytic leukemia. In the pre-ATRA era pulmonary bleeding was a common cause of death in these patients, mostly due to disseminated intravascular coagulation which was further exacerbated by the administration of chemotherapy. Although ATRA syndrome, the most serious adverse effect of ATRA treatment, involves the lungs, pulmonary hemorrhage has only rarely been reported as a manifestation of ATRA syndrome. Here we describe 2 patients who developed diffuse alveolar hemorrhage during treatment with ATRA. The possible mechanisms of pulmonary bleeding in these cases are discussed. PMID- 11042520 TI - Epstein-Barr virus (EBV)-induced B-cell proliferative disorder after chemotherapy in a patient with hemophagocytic lymphohistiocytosis with associated EBV-induced T-cell proliferation. AB - We report a case of Epstein-Barr virus (EBV)-associated lymphoproliferative disorder (LPD) which developed after chemotherapy for hemophagocytic lymphohistiocytosis (HLH), who had no history of immunodeficiency or familial X linked LPD. In HLH, the presence of EBV in T-cells was confirmed by a combination of in situ hybridization (ISH) and immunostaining. Southern blot analysis using EBV-TR and immunoglobulin JH probes revealed oligoclonal proliferation of B-cells in each organ involved by abnormal B-lymphoid cells at autopsy. Combined ISH and immunostaining disclosed the presence of EBV in proliferating B-cells. Cytokine analysis during the period of T-cell activation in HLH revealed marked elevation of interferon (IFN) gamma, interleukin (IL)-10 and soluble IL-2 receptor (sIL-2R) and mild to moderate increases of tumor necrosis factor (TNF)-alpha were observed, while IFN gamma, IL-10 and sIL-2R were elevated initially during the HLH phase, which then decreased as LPD developed and B-cell proliferation predominated. Immunosuppressive chemotherapy for HLH may then have allowed latent EBV in B lymphocytes to induce transformation and oligoclonal proliferation of B cells, finally resulting in LPD. Mechanisms of EBV-induced cell proliferation remain unclear, but alteration of various cytokines may be responsible for it. PMID- 11042522 TI - Breast plasmacytoma. AB - An unusual case of breast plasmacytoma associated with a multiple myeloma is reported. Breast plasmacytoma is exceedingly rare. We identified 37 cases published between 1928 and 1999, and reviewed the clinical features of this unusual presentation of plasmacytoma. PMID- 11042523 TI - Chronic myelomonocytic leukemia derived from a possible common progenitor of monocytes and natural killer cells. AB - The neural cell adhesion molecule, CD56, is expressed on acute myelogenous leukemia (AML) cells in 17-20% of the patients. However, the clinical and biological significance of its expression in AML has not been well analyzed from the standpoint of CD56 expression and its association with differentiation to a natural killer (NK) cell lineage. Here we present a 78-year-old patient with chronic myelomonocytic leukemia (CMML) whose leukemic cells had features of both monocytes and NK cells. We demonstrated that the leukemic cells were positive for CD4, CD56 and interleukin-2 (IL-2) receptor beta chain (CD112) in addition to myelomonocytic markers such as CD33, CD11b and CD11c. These leukemic cells proliferated well in vitro in response to 10-100 U/ml of IL-2, and functionally showed significant cytotoxicity against K562 target cells in a 4-hour (51) Cr release assay. All the above data indicate that these cells possessed at least some of the biological features of NK cells. Accordingly, we speculate that the leukemic cells in this patient may have been derived from a possible common progenitor of monocytes and NK cells. PMID- 11042524 TI - Acquired delta-storage pool deficiency associated with idiopathic myelofibrosis. AB - A 73-year-old woman complained of easy bruising, as a consequence of prolonged bleeding time despite normal platelet counts. Platelet aggregation profile, mepacrine fluorescence test, flow cytometry and transmission electron microscopy studies led to the diagnosis of delta-storage pool deficiency (SPD) A few months later, she developed hyperleucocytosis with immature granulocytes and erythroblasts. The presence of bone marrow fibrosis and clonal cytogenetic abnormalities led to the diagnosis of idiopathic myelofibrosis (IM). Association between SPD and IM has never been reported. The pathogenesis of this unusual association remains unclear and may involve proliferation of abnormal monoclonal stem cells with differentiation into activated megakaryocytes associated with impaired dense granule development and increased cytokines release which may be. involved in myelofibrosis. PMID- 11042525 TI - Hodgkin's disease with primary manifestation in the liver. AB - We describe a patient with HD presenting with hepatomegaly and impaired liver function and discuss the differential diagnosis and histologic diagnosis of HD in the liver. PMID- 11042526 TI - Involvement of the appendix in a relapsed case of primary nasal NK/T-cell lymphoma. AB - We report here a 20-year-old man presenting with primary nasal NK/T-cell lymphoma which showed an aggressive clinical course spreading to the spleen and skin despite various treatments. Eight months after high dose chemotherapy followed by autologous peripheral blood stem cell transplantation, acute appendicitis with perforation occurred and the patient underwent appendectomy. The histopathological diagnosis was NK/T-cell lymphoma of the appendix. Lymphoma of the appendix is extremely rare and the majority of appendiceal lymphomas are of B cell origin. This is the first report of involvement of appendix by nasal NK/T cell lymphoma. PMID- 11042527 TI - Avascular necrosis of head of femur in a patient with acute promyelocytic leukemia. AB - Avascular necrosis (AVN) of head of the femur is associated with various pathological conditions and treatment modalities. We present a case of acute promyelocytic leukemia who was treated with all-transretinoic acid (ATRA), daunomycin, cytarabine and a short course of dexamethasone. He developed AVN of bone after 2 years of treatment. Whether this is related to ATRA is dealt with in the discussion. PMID- 11042528 TI - Divergent expression of cyclin-dependent kinase inhibitors (CKI) and p14ARF/p16 beta in non-Hodgkin's lymphomas and chronic lymphocytic leukemia. AB - Chronic B-cell lymphocytic leukaemia (CLL) and low-grade B-cell Non Hodgkin's lymphomas (Lg-NHL) are characterized by slow accumulation of neoplastic cells arrested in the G0/G1 phase of the cell cycle. In contrast, proliferation rates are high in aggressive B-cell lymphomas (Hg-NHL). Divergent expression of cyclin dependent kinase inhibitors (CKI) in the cell cycle may contribute to these differences. We analysed CLL as well as low and high grade B-cell NHL for expression of G1-specific and universal CKI by competitive RT-PCR and immunostaining. p16(INK4A) expression was low in all types of neoplasms. Highest p14(ARF) /p16 beta expression levels were found in normal lymphocytes. Expression of this CKI was significantly lower in CLL, but still higher in CLL than in the lymphomas (median 27 vs. 3 mRNA transcripts x 10(3), p = 0.0001). p14(ARF) /p16 beta immunostaining correlated with mRNA expression. Highest p21 mRNA levels were found in CLL, but three of four CLL with abundant p21 mRNA production were negative on immunostaining. High grade lymphomas showed markedly decreased p21 expression (3.9 in Hg-NHL vs. 12 in Lg-NHL and 29 in CLL; values expressed as mRNA transcripts x 10(3), p < 0.009). mRNA and protein expression of p27 was considerably higher in CLL than in the lymphomas. Differential CKI expression in various B-cell neoplasias may provide important biological markers, if not the molecular underpinning of their different cell cycle kinetics. Targeted interference with such genes governing cell cycle control in lymphoid neoplasia may pave the way towards new treatment strategies. PMID- 11042529 TI - Arsenic trioxide and the growth of human T-cell leukemia virus type I infected T cell lines. AB - A novel therapeutic potential for acute promyelocytic leukemia using arsenic trioxide (As(2) O(3) ) has been reported. Recent in vitro studies demonstrated that As(2) O(3) effectively inhibits the growth of some cell lines derived from patients with malignant lymphoma, chronic lymphocytic leukemia and multiple myeloma. Adult T-cell leukemia (ATL) is an aggressive neoplasm of mature T-cell origin caused by human T-cell leukemia virus type-I (HTLV-I) the prognosis of which still remains very poor. A possible role of As(2) O(3) for the treatment of ATL is demonstrated from evidence that As(2) O(3) significantly inhibits the growth of HTLV-I infected T-cell lines and induces apoptosis in fresh ATL cells at clinically achievable concentration of the agent. The growth inhibition of As(2) O(3) treated HTLV-I infected T-cell lines was induced by both apoptosis and G(1) phase accumulation. Cleaved bcl-2 protein and an enhanced expression of bak protein in the cells were coincidentally observed during As(2) O(3) treatment. A broad spectrum caspase inhibitor, z-Val-Ala-DL-Asp-fluoromethylketone inhibited the apoptosis induced by As(2) O(3). Increased expression of p53, Cip1/p21 and Kip1/p27, and dephosphorylation of retinoblastoma protein (pRb) were detected in the As(2) O(3) treated cells. In conclusion, As(2) O(3) might become a new therapeutic tool in the treatment of ATL as As(2) O(3) induces apoptosis by destruction of the bcl-2 protein and enhancement of the bak protein production proceeding to activate caspases, and also induces G(1) phase accumulation by enhancement of p53, Cip1/p21, Kip1/p27 and dephosphorylation of pRb to HTLV-I infected T-cell lines. PMID- 11042530 TI - B-cell leukemia in a rhesus macaque (Macaca mulatta) infected with simian immunodeficiency virus. AB - Conditions associated with abnormal B-cell proliferation have an increased incidence in the HIV-infected population. A longitudinal study conducted at the Tulane Regional Primate Research Center has followed more than 1,000 rhesus macaques infected with simian-immunodeficiency virus (SIV) since 1984. While spontaneous B-cell malignancy in SIV-negative macaques has not been reported, 42 cases of SIV-associated-lymphoma (SAL) have been documented in this cohort. Recently we identified a single case of B-cell leukemia, first suggested by clinical abnormalities and confirmed and further characterized by molecular analysis. The case is important because it models the occurrence of B-cell leukemia in the human AIDS patient and because it extends our understanding of the B-cell lymphoproliferative diseases associated with AIDS. PMID- 11042531 TI - Effect of ubenimex (Bestatin) on the cell growth and phenotype of HL-60 and HL 60R cell lines: up-and down-regulation of CD13/aminopeptidase N. AB - Ubenimex (Bestatin), a low-molecular-mass dipeptide, has been demonstrated to have anti-tumor activities and immunomodulating activities. We here report cell growth inhibition and phenotypic changes of HL-60 and HL-60R cell lines induced by Bestatin treatment. Bestatin (0.1 microg/ml) showed remarkable cell growth inhibition against HL-60 cells, whereas it was ineffective for HL 60R cells. Bestatin also showed growth inhibition in the concentration of 1 microg/ml against HL-60R cells which are resistant to differentiation induction by DMSO and TPA. In both cell types, the effect of growth inhibition by Bestatin treatment was dose dependent. We found a low level expression of CD13 and a low number of CD13 positive cells in HL-60R cells compared with that of HL-60. We also observed phenotypic changes of HL-60 and HL-60R cells following incubation with Bestatin (10 microg/ml) for 1 and 3 hrs, respectively. With HL-60 cells, the upregulation of CD13/aminopeptidase N was found after 1 hr, however, the downregulation was observed after 3 hrs incubation with Bestatin. On the other hand, the downregulation of CD15 and CD33 was observed after both one and 3 hrs incubation. Similarly, in HL-60R cells, the upregulation of CD13/aminopeptidase N was found temporarily (1hr), and then CD13 downregulation was observed after 3 hrs incubation with Bestatin. No notable change was observed for expression of other myeloid-related antigens, e.g. CD14 (My4, LeuM3), CD11b (OKM1), and CD34 (My10). On the basis of these observations of in vitro activity, we suggest that Bestatin may also be an effective anti-leukemic agent in vivo. PMID- 11042532 TI - A view on the science: physical anthropology at the millennium. PMID- 11042533 TI - Molecular evidence for different stages of tuberculosis in ancient bone samples from Hungary. AB - This paleomicrobiologic study was conducted on osseous tissue specimens from ancient Hungarian skeletal samples from the 7-8th and the 17th centuries AD with typical macromorphologic evidence of osseous tuberculosis (n = 3), morphologic alterations probably due to tuberculosis (n = 6), or with nontypical osseous changes of vertebral bodies suggestive of inflammatory reaction (n = 5). From these bone samples, DNA was extracted and amplified by polymerase chain reaction (PCR) by using various primer pairs recognizing DNA segments of different mycobacterial species. To confirm specificity of the analysis, the amplification products of several samples were subjected to restriction enzyme digestion and/or direct sequencing. Of the analyzed 14 cases, 8 were unambiguously positive for mycobacterial DNA of the Mycobacterium tuberculosis complex, as shown by the amplification of the IS6110 sequence. In 13 cases we found a PCR product with primers specific for the 65-kDa antigen gene, including 2 cases without genomic DNA. We conclude that the application of other mycobacterial DNA primers may reveal contamination of bones with atypical saprophytic mycobacteria. A positive result for typical mycobacteria was seen in 2 of 3 cases with typical morphologic signs of tuberculosis and amplifiable DNA, in 3 of 6 probable cases, but also in 3 of 6 cases with nontypical bone changes. This indicates that minor osseous reactions of the surface of vertebral bodies may be due-at least in several cases to infections with bacteria of the M. tuberculosis complex. In these cases the disease may have proceeded rapidly, and the morphologic osseous changes may represent "early" stages of tuberculous infection of the vertebrae. PMID- 11042534 TI - Fetal age: methods of estimation and effects of pathology. AB - Accurate identification of fetal age is important in a wide variety of circumstances. Seventeen anthropometric and radiographic measurements were taken on fetuses between 15 and 42 weeks of gestational age, both with and without pathologic conditions. A full evaluation including radiographic, karyotypic, gross anatomic, and histologic examination of the fetus and placenta identified 72 individuals as nondysmorphic with no signs of chronic uterovascular insufficiency. These specimens served as the control group. Based on least squares regressions of this group, age-estimation equations were calculated for all variables. Six models were adequately described by linear equations; the remaining 11 required a quadratic term. Based on standard error of the estimate (S(y:x)), skeletal measures proved the most accurate age estimators. Pathologic conditions were shown to have an influence on age estimation indicated by high levels of inaccuracy and, in some instances, significant bias. PMID- 11042535 TI - Sex assessment on the basis of long bone circumference. AB - Discriminant functions have long been used to classify individuals into groups according to the dimensions of their bones. Although lengths, widths, and diameters have been extensively used, the circumferences have not been adequately validated. In this work, the importance that the circumferences of long bones can have in assigning the sex of ancient human remains is demonstrated. The functions produced by using just one circumference achieved accuracies higher than 80%, and circumference at the radial tuberosity of the radius is able to classify 92.8% of skeletons from the Late Roman site of Mas Rimbau/Mas Mallol (Spain). When functions are produced by using more than one circumference, they can achieve the uppermost classification attained in this sample. The functions also showed that the arm circumference functions are more useful than those of the leg, probably because male individuals of the population had greater mechanical stress than did females. The classification percentages, as well as other statistical values for the functions, demonstrated the great ability of long bone circumferences in helping to classify the sex of individuals of other sites of the Mediterranean area besides the ones examined in this study. PMID- 11042536 TI - Shape variation of the human pollical distal phalanx and metacarpal. AB - Human distal pollical phalanx form has been associated with tool manufacture, and the broad tuft of this bone in Neanderthals has been suggested to be a climatic adaptation and/or an aid to a tremendously powerful grip. A wide first metacarpal head has also been proposed to be useful in distinguishing tool-dependent hominids from those less reliant on tools. In order to contribute to an evaluation of these hypotheses variation in first metacarpal and distal phalanx shape is explored among samples of modern humans and compared to that of fossil hominids. Modern humans are from the Terry Collection, Larsen Bay, a Chinese Alaskan cemetery, Egypt, and Sully and Mobridge. Hominid fossils include AL 333w 39, SKX 5016, SK 84, Stw 294, OH 7, several Neanderthals, Skhul 4 and 5, and Predmosti 3. Analysis involves length-width ratios, regressions of distal phalanx tuft width on base width and of metacarpal head width on length, and pattern profiles based on Z-scores with reference to the Larsen Bay sample. Larsen Bay individuals are robust, while Terry "blacks," Egyptians, and Chinese-Alaskan males tend to be gracile. Fossil hominids are most distinctive for distal phalanx radioulnar tuft and mid-shaft widths relative to length. Security of grip is one plausible explanation. While most modern samples are positively allometric for tuft width relative to base width, the Larsen Bay and fossil hominid samples are not; thus caution is advised in accepting a base-tuft width comparison as a tool dependence marker. Separation from modern humans is not easily achieved with metacarpal measures, but the Hadar metacarpal has distinctively narrow radioulnar head width ratios. While first metacarpal head expansion among hominids may plausibly be related to tool manufacture, other activities that place stress on the metacarpophalangeal joint should also be considered. PMID- 11042537 TI - The value of infracranial nonmetric variation in studies of modern Homo sapiens: an Australian focus. AB - The value of quantitative infracranial nonmetric variation is examined in the study of population relationships by using samples from populations originating from five major geographic regions: Australia (two populations), Africa, East Asia, Europe, and Polynesia. According to the nonspecificity hypothesis, there are no distinct large classes of genes affecting one group of attributes exclusively; thus infracranial nonmetric traits should compare with other osteologic data sets in addressing questions of population relationships. By using the mean measure of divergence, infracranial nonmetric traits are shown to be useful in separating populations, particularly when using female and pooled sex samples. The two Australian female samples (New South Wales coastal Australian and South Australian Aboriginals) are shown to be closer than any other two samples. The picture of intrapopulation and interpopulation variation in infracranial nonmetric traits is extended and clarified. Distance studies with infracranial nonmetric traits are possible but more illuminating if the sexes are first separated. Infracranial nonmetric variation does extend the knowledge of human population studies in yielding biologically meaningful results relating to development and ontogeny. PMID- 11042538 TI - Fourier analysis of facial profiles of young twins. AB - Twins studies provide a powerful approach to determining the relative contribution of genetics and environment to observed variation. Such studies assume trait differences in monozygous (MZ) twins are due to environmental factors and those in dizygous (DZ) twins are due to both genetic and environmental factors. This study quantitated facial profiles of twins using Fourier equations, determining their value in profile analysis and the assessment of the genetic contribution to facial shape. Standardized profile slide photographs of 79 pairs of 4-6 year-old twins (37 MZ pairs, 42 DZ pairs) were scanned and x and y coordinates were extracted from each profile using sellion and Camper's plane as references. The coordinates were subjected to Fourier analysis and the normalised vertex projection coefficients were studied. The means of the differences between coefficients for MZ co-twins did not differ significantly from that of DZ co-twins, although the DZ group showed higher mean differences in the higher harmonics. Subjective examination of superimposed reconstructions showed wider variation between DZ co-twins than MZ co-twins. Correct classification of twins by discriminant function analysis using Fourier coefficients was similar for both groups (MZ: 70.3%; DZ: 73.8%). Fourier analysis could quantitate facial profiles of young children and differentiate some details, but was unable to discriminate between genetic and environmental influences, and any possible interactions between these influences, on their overall facial profiles at this developmental stage. PMID- 11042539 TI - Differences of subcutaneous adipose tissue topography in type-2 diabetic (NIDDM) women and healthy controls. AB - Women suffering from type-2 diabetes mellitus (non-insulin-dependent diabetes mellitus [NIDDM]) have more total body fat and upper body obesity compared with healthy controls. However, the standard measurement methods have disadvantages such as radiological burden, lack of precision, or high time consumption. A new optical device, the Lipometer, enables the noninvasive, quick, and save determination of the thickness of subcutaneous adipose tissue layers at any given site of the human body. The specification of 15 evenly distributed body sites allows the precise measurement of subcutaneous body fat distribution, so-called subcutaneous adipose tissue topography (SAT-Top). SAT-Tops of 20 women with clinically proven NIDDM and 122 healthy controls matched by age group were measured. In this paper, we describe the precise SAT-Top differences of these two groups and present the multidimensional SAT-Top information condensed in a two dimensional factor plot and in a response plot of an artificial neural network. NIDDM women provide significantly lower leg SAT-Top and significantly higher upper trunk SAT-Top development ("apple"-type) compared with their healthy controls. PMID- 11042540 TI - Bone mineral density, osteopenia, and osteoporosis in the rhesus macaques of Cayo Santiago. AB - This cross-sectional study investigates metabolic bone disease and the relationship between age and bone mineral density (BMD) in males and females of a large, well-documented skeletal population of free-ranging rhesus monkeys (Macaca mulatta), from the Caribbean Primate Research Center Museum collection from Cayo Santiago, Puerto Rico. The sample consists of 254 individuals aged 1.0-20+ years. The data consist of measurements of bone mineral content and bone mineral density, obtained from dual-energy X-ray absorptiometry (DEXA), of the last lumbar vertebra from each monkey. The pattern of BMD differs between male and female rhesus macaques. Females exhibit an initial increase in BMD with age, with peak bone density occurring around age 9.5 years, and remaining constant until 17.2 years, after which there is a steady decline in BMD. Males acquire bone mass at a faster rate, and attain a higher peak BMD at an earlier age than do females, at around 7 years of age, and BMD remains relatively constant between ages 7-18.5 years. After age 7 there is no apparent effect of age on BMD in the males of this sample; males older than 18.5 years were excluded due to the presence of vertebral osteophytosis, which interferes with DEXA. The combined frequency of osteopenia and osteoporosis in this population is 12.4%. BMD values of monkeys with vertebral wedge fractures are generally higher than those of virtually all of the nonfractured osteopenic/osteoporotic individuals, thus supporting the view that BMD as measured by DEXA is a useful but imperfect predictor of fracture risk, and that low BMD may not always precede fractures in vertebral bones. Other factors such as bone quality (i.e., trabecular connectivity) should also be considered. The skeletal integrity of a vertebra may be compromised by the loss of key trabeculae, resulting in structural failure, but the spine may still show a BMD value within normal limits, or within the range of osteopenia. PMID- 11042541 TI - Craniometric variation in a population of mantled howler monkeys (Alouatta palliata): evidence of size selection in females and growth in dentally mature males. AB - A large body of work on monkey cranial metrics (involving conclusions about interspecific variation, sexual dimorphism, and ontogeny) depends on the assumptions that growth effectively ceases with dental maturity and that intraspecific variation is negligible. We test these assumptions by examining variation in 39 measurements of 166 dentally mature Alouatta palliata skulls from animals found dead on Barro Colorado Island (BCI), Panama. We also investigate whether this population is under size-based selection, since our found-dead sample reflects the natural mortality in this population. The sample was divided into three age stages by occlusal wear (A-C, least to most wear). Female stage A means are significantly smaller than female stage B means for three cranial measures. Female stage B means are significantly smaller than female stage C means for five cranial measures. Male stage A means are significantly smaller than male stage B means for 21 cranial measures. Multivariate analyses confirm this trend of expansion between adult age stages. The dental metric and suture closure data suggest that the cranial expansion in females is due to size-based selection, while the cranial expansion in males is due to significant growth after dental maturity. Sexual dimorphism ratios are highly variable across different samples of A. palliata, indicating that dimorphism varies between populations of this species. These results provide insight into the selective forces operating on the BCI howlers and challenge the validity of the many studies which pool subspecies and assume growth ceases with maturity. PMID- 11042542 TI - Grandmother hypothesis and primate life histories. AB - The adaptive significance of midlife menopause in human females has long engaged the attention of evolutionary anthropologists. In spite of extensive debate, the problem has only recently been examined in the context of primate life histories. Here I extend those investigations by comparing life history traits in 16 primate species to test predictions generated from life history theory. In humans, late ages of maturity and higher than expected birth rates are systematically associated with extended postmenopausal longevity. Links among these adjustments on the primate pattern can explain how selection could slow somatic senescence without favoring extension of the fertile span. This conclusion is consistent with the observation that our fertile spans are similar to those of other pongids. The shape of the argument herein demonstrates the utility of life history theory for solving problems of adaptive evolution in female life history traits, with consequences for broader arguments regarding human evolution. PMID- 11042543 TI - Cell encapsulation with alginate and alpha-phenoxycinnamylidene-acetylated poly(allylamine). AB - In general, microcapsules prepared from alginate and polycations lack mechanical strength because the interaction between alginate and polycations is ionic instead of covalent, which represents a much stronger bond. To increase the mechanical strength of the capsule, we prepared photosensitive microcapsules that could form covalent bonds between polymers in the capsular membrane by light irradiation. Two types of photosensitive poly(allylamine), with 5% and 10% of amino groups modified by alpha-phenoxycinnamylidene acetylchloride, were synthesized. Both photopolymers exhibited an absorption maximum at 325 nm and were capable of crosslinking upon light exposure. These photosensitive polymers were used for the preparation of microcapsules. The capsules formed from this photosensitive poly(allylamine) and alginate were strengthened significantly by light irradiation. Only 28% of the microcapsules prepared from the 5%-modified photopolymer fractured after 48 h of shaking at 150 rpm. This fracture percentage is much lower when compared with the 60% of capsules fractured when prepared from the untreated poly(allylamine). By using poly(allylamine) at 10% modification, the mechanical strength was improved only slightly, with 26% of capsules fractured. Analysis of the permeability test indicated that the photo-crosslinked capsular membrane was freely permeable to cytochrome c and myoglobin, but less permeable to serum albumin. The encapsulation method was used to entrap and culture IW32 mouse leukemia cells. The cells proliferated to a density of about 1.1 x 10(7) cells/mL in the capsules after 7 days of cultivation. Concurrently, the concentration of erythropoietin in the microcapsules increased to 800 mU/mL. This new encapsulation technique has great potential in the application of a bioindustrial cell-culturing process. PMID- 11042544 TI - Separation of alpha-acid glycoprotein glycoforms using affinity-based reversed micellar extraction and separation. AB - A new method for the preparation of the glycoforms of bovine alpha(1)-acid glycoprotein (AGP) is described relying on affinity-reversed micellar extraction and separation (ARMES). This method has proven effective in separating structurally similar glycoproteins and separating glycoproteins from nonglycosylated proteins from natural sources. In this method, individual glycoforms complex with the lectin, concanavalin A (ConA) are extracted into an organic-phase reversed micellar solution formed by Aerosol OT (AOT). The purity of three AGP glycoforms isolated was assessed by hydroxyapatite high-performance liquid chromatography (HPLC), gel-permeation chromatography and SDS-PAGE. The glycan structure of the pure glycoforms was analyzed. Oligosaccharide mapping using capillary electrophoresis (CE) and PAGE showed the glycans obtained from each glycoform to be distinctly different. ARMES can be used for the semi preparative scale resolution of the glycoforms of bovine AGP or other therapeutic glycoproteins. PMID- 11042545 TI - Toxicity effects of compressed and supercritical solvents on thermophilic microbial metabolism. AB - Selection of biocompatible solvents is critical when designing bioprocessing applications for the in situ biphasic extraction of metabolic end-products. The prediction of the biocompatibility of supercritical and compressed solvents is more complicated than for liquid solvents, because their properties can change significantly with pressure and temperature. The activity of the anaerobic thermophilic bacterium, Clostridium thermocellum, was studied when the organism was incubated in the presence of compressed nitrogen, ethane, and propane at 333 K and multiple pressures. The metabolic activity of the organisms in contact with compressed solvents was analyzed using traditional indicators of solvent biocompatibility, such as log P, interfacial tension, and solvent density. The toxicity of the compressed solvents was compared with the phase and molecular toxicity effects measured in liquid alkanes at atmospheric pressure. Inactivation increased with time in the presence of the compressed solvents, but was constant in the presence of atmospheric liquid solvents. Knowledge of molecular and phase toxicity provides a framework for the interpretation of C. thermocellum metabolism in contact with atmospheric and compressed solvents. PMID- 11042546 TI - Phase equilibria in the lysozyme-ammonium sulfate-water system. AB - Ternary phase diagrams were measured for lysozyme in ammonium sulfate solutions at pH values of 4 and 8. Lysozyme, ammonium sulfate, and water mass fractions were assayed independently by UV spectroscopy, barium chloride titration, and lyophilization respectively, with mass balances satisfied to within 1%. Protein crystals, flocs, and gels were obtained in different regions of the phase diagrams, and in some cases growth of crystals from the gel phase or from the supernatant after floc removal was observed. These observations, as well as a discontinuity in protein solubility between amorphous floc precipitate and crystal phases, indicate that the crystal phase is the true equilibrium state. The ammonium sulfate was generally found to partition unequally between the supernatant and the dense phase, in disagreement with an assumption often made in protein phase equilibrium studies. The results demonstrate the potential richness of protein phase diagrams as well as the uncertainties resulting from slow equilibration. PMID- 11042547 TI - Formaldehyde-mediated aggregation of protein antigens: comparison of untreated and formalinized model antigens. AB - A formaldehyde-mediated aggregation pathway (FMAP) is suggested as being primarily responsible for the aggregation of lyophilized tetanus toxoid (TT; a formalinized antigen) in the presence of moisture. The general occurrence of the FMAP was examined by using bovine serum albumin (BSA) and ribonuclease A (RNase) as model antigens; both protein antigens were formalinized according to a method commonly used to detoxify bacterial toxins. To clearly delineate the FMAP from other aggregation mechanisms, the aggregation kinetics and mechanism of both unmodified antigens (BSA and RNase) and formalinized antigens (f-BSA and f-RNase) were evaluated. We report that formaldehyde treatment introduces more rapid and extensive aggregation in antigens under conditions that favor the FMAP (i.e., 80% relative humidity and 37 degrees C). Consistent with formaldehyde-mediated crosslinking, f-antigen aggregates were covalent and non-disulfide-bonded, whereas BSA aggregates were disulfide-linked and RNase even did not aggregate under the same conditions. Coincorporation of amino acids (histidine and lysine), which strongly interact with formaldehyde, as well as prior antigen reduction with cyanoborohydride, significantly inhibited f-BSA aggregation, but showed no selective effect on BSA aggregation. Mechanistic analysis of f-BSA aggregates, inhibition studies, and similar reactivity of f-BSA with TT all confirmed the existence of the FMAP at moisture levels intermediate between the dry and solution state. This study demonstrates the potential for covalent reactions between formalinized protein antigens and neighboring chemical or biochemical species even after formalinization, and provides a general approach to inhibit the FMAP. PMID- 11042548 TI - Enhanced bioaccumulation of heavy metals by bacterial cells displaying synthetic phytochelatins. AB - A novel strategy using synthetic phytochelatins is described for the purpose of developing microbial agents for enhanced bioaccumulation of toxic metals. Synthetic genes encoding for several metal-chelating phytochelatin analogs (Glu Cys)(n)Gly (EC8 (n = 8), EC11 (n = 11), and EC20 (n = 20)) were synthesized, linked to a lpp-ompA fusion gene, and displayed on the surface of E. coli. For comparison, EC20 was also expressed periplasmically as a fusion with the maltose binding protein (MBP-EC20). Purified MBP-EC20 was shown to accumulate more Cd(2+) per peptide than typical mammalian metallothioneins with a stoichiometry of 10 Cd(2+)/peptide. Cells displaying synthetic phytochelatins exhibited chain-length dependent increase in metal accumulation. For example, 18 nmoles of Cd(2+)/mg dry cells were accumulated by cells displaying EC8, whereas cells exhibiting EC20 accumulated a maximum of 60 nmoles of Cd(2+)/mg dry cells. Moreover, cells with surface-expressed EC20 accumulated twice the amount of Cd(2+) as cells expressing EC20 periplasmically. The ability to genetically engineer ECs with precisely defined chain length could provide an attractive strategy for developing high affinity bioadsorbents suitable for heavy metal removal. PMID- 11042549 TI - Enhancement of mass transfer using colloidal liquid aphrons: measurement of mass transfer coefficients in liquid-liquid extraction. AB - Interphase mass transfer of a sparingly soluble solute is often the rate-limiting step in multiphase biocatalytic processes. Colloidal liquid aphrons (CLA) provide very large interfacial areas, and thus could enhance mass transfer in such processes. The aim of this study was to characterize mass transfer properties of CLA dispersions during transfer of heptanoic acid from water to limonene. The interfacial area per unit volume (a), film mass transfer coefficient (K(L)), and volumetric mass transfer coefficient (K(L)a) values were determined in a stirred tank reactor. These results were used, along with a literature correlation, to estimate the mass transfer coefficient of the surfactant-stabilized shell surrounding the CLA. The very large increase in a provided by the CLA was only partially offset by a slight increase in the mass transfer resistance of the shell. As a result, the range of K(L)a values obtained using CLA was about an order of magnitude greater than that obtained using a conventional dispersion. The concentration of the aqueous-phase surfactant used to form the CLA strongly affected the Sauter mean diameter of the CLA; however, the concentration of the nonpolar-phase surfactant had little effect. These results suggest that CLA have considerable potential for multiphase biocatalytic applications. PMID- 11042550 TI - Ethanol utilization by sulfate-reducing bacteria: an experimental and modeling study. AB - A mixed culture of sulfate-reducing bacteria containing the species Desulfovibrio desulfuricans was used to study sulfate-reduction stoichiometry and kinetics using ethanol as the carbon source. Growth yield was lower, and kinetics were slower, for ethanol compared to lactate. Ethanol was converted into acetate and no significant carbon dioxide production was observed. A mathematical model for growth of sulfate-reducing bacteria on ethanol was developed, and simulations of the growth experiments on ethanol were carried out using the model. The pH variation due to sulfate reduction, and hydrogen sulfide production and removal by nitrogen sparging, were examined. The modeling study is distinct from earlier models for systems using sulfate-reducing bacteria in that it considers growth on ethanol, and analyzes pH variations due to the product-formation reactions. PMID- 11042551 TI - Determination of depolymerization kinetics of amylose, amylopectin, and soluble starch by Aspergillus oryzae alpha-amylase using a fluorimetric 2-p toluidinylnaphthalene-6-sulfonate/flow-injection analysis system. AB - This study reports on the determination of the depolymerization kinetics of amylose, amylopectin, and soluble starch by Aspergillus oryzae alpha-amylase using flow-injection analysis with fluorescence detection and 2-p toluidinylnaphthalene-6-sulfonate as the fluorescent probe. The experimental data points, corresponding to the evolution of the concentration of "detectable" substrate with depolymerization time, were fit to a single exponential decay curve in the case of amylose and to a double exponential decay curve in the cases of amylopectin and soluble starch. For all the assayed substrates, the determined depolymerization rates at time zero correlated well with the initial enzyme and substrate concentrations through the usual Michaelis-Menten hyperbola. Therefore, this methodology allows the determination of alpha-amylase activity using any of these substrates. For amylopectin and soluble starch, the value of the total depolymerization rate at any depolymerization time was the result of the additive contribution of two partial depolymerization rates. In contrast, the total depolymerization rate for amylose was always a single value. These results, in conjunction with the relative time evolution of the two partial depolymerization rates (for amylopectin and soluble starch), are in good agreement with a linear molecular structure for amylose, a "grape-like" cluster molecular structure for amylopectin, and an extensively degraded grape-like cluster structure for soluble starch. PMID- 11042552 TI - Bioconversion of hydrophobic compounds in a continuous closed-gas-loop bioreactor: feasibility assessment and epoxide production. AB - Microorganisms can be used as catalysts to produce organic compounds in a highly chemo-, regio- and enantioselective manner, and whole cells do not require the costly addition of cofactors for redox reactions. However, bioconversions are slow compared to alternative chemical reactions, and the biocatalyst works at its best in an aqueous medium, while the transformations of interest frequently involve compounds with a low-aqueous solubility and that are toxic to microorganisms. This results in low-volumetric productivity in classical bioreactors. The Continuous Closed-Gas-Loop Bioreactor is described here-a reactor system with high productivity, but without the problems associated with two-phase systems, such as an emulsified product stream and phase toxicity. Its working principle is to recirculate a gas phase through a bioreaction compartment and a saturator/absorber module where the product accumulates as a clear organic solution. A wide range of bioconversions should be possible in this set-up, and proof of concept was established for the epoxidation of 1,7-octadiene to (R)-1,2 epoxyoct-7-ene by a native strain of Pseudomonas oleovorans. This reaction represents a group of terminal alkene epoxidations where the bioconversion substrate does not support growth of the microorganism. Practical results at a 5l scale are presented for this bioconversion for both batch and continuous operation with respect to the aqueous phase, showing continuous stable epoxidation at productivities >14 micromol min(-1) L(-1) (U L(-1)). The results confirm that the metabolism does not allow a simple optimization strategy, because growth and biotransformation substrates compete for the same enzyme sites, and conversely growth on a substrate using this very enzyme system is necessary for longterm bioconversion. Integrated removal of the CO(2) formed via the liquid overflow was estimated from theory and verified in experimental work. PMID- 11042554 TI - Perfusion culture of baculovirus-infected BTI-Tn-5B1-4 insect cells: a method to restore cell-specific beta-trace glycoprotein productivity at high cell density. AB - The impact of different cultivation-infection strategies on the productivity of baculovirus-infected BTI-Tn-5B1-4 (High Five) cells was investigated. Using beta trace protein as the recombinant glycoprotein, the effects of multiplicity of infection (MOI) and time of infection (TOI) were studied on growth after infection as well as the degree of infection and recombinant protein productivity in batch culture. The highest productivities were found when infecting Tn5 cells at early exponential growth phase (EGP) (low cell density) using a high MOI. To increase the productive cell density of Tn5 cells after beta-trace-baculovirus infection, we performed studies infecting cells in the range of 1 to 5 x 10(6) cells/mL in fresh medium. Although the protein production was increased twofold, a strong negative cell density effect was still observed when maximal productive cell density exceeded 1 x 10(6) cells/mL. To verify whether the changing cell environment of the batch experiments was responsible for the decrease in protein productivity at increasing cell density at infection, several perfusion experiments were designed by infecting Tn5 cells at cell densities over 2 x 10(6) cells/mL under more steady-state conditions. The use of this experimental setup enabled successful infections at high cell densities with volumetric productivities of up to 1.2 g L(-1) day(-1) of beta-trace protein, which is very high for a glycoprotein expressed with the baculovirus expression vector system (BEVS). The cell specific protein productivity observed after infections at higher cell densities in perfusion mode was the same as in batch experiments at low cell concentrations, which clearly demonstrates that the cell density effect could be completely overcome with perfusion cultivation. PMID- 11042553 TI - Enzymatic grafting of hexyloxyphenol onto chitosan to alter surface and rheological properties. AB - An enzymatic method to graft hexyloxyphenol onto the biopolymer chitosan was studied. The method employs tyrosinase to convert the phenol into a reactive o quinone, which undergoes subsequent nonenzymatic reaction with chitosan. Reactions were conducted under heterogeneous conditions using chitosan films and also under homogeneous conditions using aqueous methanolic mixtures capable of dissolving both hexyloxyphenol and chitosan. Tyrosinase was shown to catalyze the oxidation of hexyloxyphenol in such aqueous methanolic solutions. Chemical evidence for covalent grafting onto chitosan was provided by three independent spectroscopic approaches. Specifically, enzymatic modification resulted in (1) the appearance of broad absorbance in the 350-nm region of the UV/vis spectra for chitosan films; (2) changes in the NH bending and stretching regions of chitosan's IR spectra; and (3) a base-soluble material with (1)H-NMR signals characteristic of both chitosan and the alkyl groups of hexyloxyphenol. Hexyloxyphenol modification resulted in dramatic changes in chitosan's functional properties. On the basis of contact angle measurements, heterogeneous modification of a chitosan film yielded a hydrophobic surface. Homogeneously modified chitosan offered rheological properties characteristic of associating water-soluble polymers. PMID- 11042555 TI - Synthesis of glycerides containing n-3 fatty acids and conjugated linoleic acid by solvent-free acidolysis of fish oil. AB - Menhaden oil, a rich source of n-3 fatty acids, was interesterified with conjugated linoleic acid (CLA) in a reaction medium composed solely of substrates and either free or immobilized commercial lipase preparations. Of five lipases tested, an immobilized preparation from Mucor miehei provided the fastest rate of incorporation of CLA into fish oil acylglycerols; however, and as observed with most of the lipases utilized, a significant proportion of the n-3 fatty acid residues were liberated in the process. A soluble lipase from Candida rugosa converted free CLA to acylglycerol residues while leaving the n-3 fatty acid residues virtually untouched. Even though the reaction rate was slower for this enzyme than for the other four lipase preparations, the specificity of the free C. rugosa lipase gives it the greatest potential for commercial use in preparing fish oils enriched in CLA residues but still retaining their original n-3 fatty acid residues. PMID- 11042556 TI - Therapeutic choices in younger patients with chronic myelogenous leukemia. AB - BACKGROUND: Allogeneic stem cell transplantation (SCT) and interferon (IFN)-alpha therapy have significantly improved the prognosis of patients with Philadelphia (Ph) chromosome positive chronic myelogenous leukemia (CML). Both therapies may be suitable for younger patients. The authors reviewed current data to assist in prioritizing these modalities in an individual patient. METHODS: The authors reviewed and summarized current data on outcomes of SCT and IFN-alpha therapy in patients with early chronic phase CML. RESULTS: Several disease-, patient-, and physician-related factors affect outcomes with both modalities. Interferon-alpha does not induce myelofibrosis. The course of CML is predictable in most patients; sudden emergence of blastic phase; disease is unusual. There is no significant adverse impact of delaying SCT for the 12 months usually necessary to assess cytogenetic response to an IFN-alpha-based regimen. Interferon-alpha may be discontinued some months before SCT and is not associated with an adverse impact on post-SCT outcomes. CONCLUSIONS: An individualized risk assessment-based approach is of value in prioritizing SCT and IFN-alpha in younger patients with chronic phase CML. The authors recommend a risk-based therapy algorithm based on the expected SCT associated 1-year mortality for an individual patient. PMID- 11042557 TI - Frey syndrome: treatment with type A botulinum toxin. AB - BACKGROUND: Frey syndrome was first described by Baillarger in 1853. Frey provided a detailed analysis and description as "auriculotemporal syndrome" in 1923. According to the literature, even the most recent therapeutic measures described for the treatment of patients with Frey syndrome have little chance of success and a high incidence of side effects. Thus, a type of treatment is desirable that can suppress the symptoms of Frey syndrome and can offer a good success rate, minimum invasiveness, and few side effects. METHODS: The experience of the authors and data from the literature confirmed the efficacy of type A botulinum toxin treatment for patients with Frey syndrome up to a maximum observation period of 3 years. RESULTS: In the current study, seven patients with severe, symptomatic Frey syndrome after parotidectomy were treated successfully with type A botulinum toxin. CONCLUSIONS: The method of local, intracutaneous treatment with type A botulinum toxin for patients with Frey syndrome is effective, virtually side-effect free, and minimally invasive. PMID- 11042558 TI - Evaluation of the validity of the 1997 International Union Against Cancer TNM classification of major salivary gland carcinoma. AB - BACKGROUND: The TNM classification (International Union Against Cancer) of salivary gland carcinoma was revised in 1997. In this study, the authors have evaluated the validity of this new TNM classification and clinical staging in 1683 patients with parotid gland carcinoma. METHODS: Reclassification was conducted according to the new classification based on the clinical data of 1683 patients registered to the salivary gland division of the Japanese Joint Committee on TNM classification. The 5- and 10-year survival rates according to TNM classification were calculated for 1074 patients whose prognosis could be followed up. Finally, the distribution of the total patient population was analyzed using the new staging and the survival curves for each disease stage. RESULTS: The variance of the patients for T1 to T4 was appropriate using the new T classification. The 5- and 10-year survival rates corresponded well to the degree of progression of TNM. However, there were only nine patients with Stage III, and marked nonuniformity in the staging was observed. The separation of the survival curves for each stage was not clear, and no significant differences between the survival curves of Stages II and III and Stages III and IV were observed. When the authors classified T1N1M0, T2N1M0, T3N1N0, and T4N0M0, which have 5- and 10-year survival rates similar to Stage III, distribution of patients and separation of the survival curves in each stage improved markedly. CONCLUSIONS: The results of the current study confirm that the new TNM classification system is valid. However, a significant problem was observed with respect to the new clinical staging. The authors propose that T1N1M0, T2N1M0, T3N1M0, and T4N0M0 be classified as Stage III. PMID- 11042559 TI - Expression of ets-1 transcription factor is correlated with penetrating tumor progression in patients with squamous cell carcinoma of the esophagus. AB - BACKGROUND: The Ets-1 transcription factor is proposed to play a role in tumor invasion. To evaluate the biologic significance of Ets-1 in the progression of squamous cell carcinoma of the esophagus, the authors investigated Ets-1 expression in tumors from 90 patients using immunohistochemical procedures. METHODS: The Ets-1 positive ratio (Ets-1 ratio) was determined in the center and deep margin areas of invasive sites and in the center and outer edge areas of intraepithelial spread contiguous with invasive disease. RESULTS: Ets-1 expression was heterogeneous within tumor tissue. The Ets-1 ratio was significantly higher at invasive sites compared with intraepithelial spread sites, and at deep margin areas compared with center areas (P < 0.0001 and P < 0.05, respectively, by two-way analysis of variance). No differences were noted in the Ets-1 ratio at the deep margin areas at each depth of invasion. Investigating the Ets-1 ratio at deep margin areas in each pathologic phenotype, the ratio of the small nest type was found to be higher than that of the large nest type (P < 0.0001). CONCLUSIONS: The authors propose that Ets-1 plays an important role in penetrating tumor progression, even very early in the invasive process (especially in the case of infiltrative invasive type tumors) at the deep margin areas of invasive sites in patients with squamous cell carcinoma of the esophagus. PMID- 11042560 TI - Serum p53 antibody is a useful tumor marker in superficial esophageal squamous cell carcinoma. AB - BACKGROUND: Patients with superficial (mucosal or submucosal) esophageal carcinoma (SEC) have significantly better survival rates than patients with advanced carcinoma. Some patients with advanced esophageal carcinoma have been reported to test positive for serum p53 antibodies (Abs). Because very few patients with superficial carcinoma have been examined, the aim of this study was to evaluate the clinical significance of serum p53-Abs in patients with superficial esophageal squamous cell carcinoma (SESCC). METHODS: Thirty-five consecutive patients with SESCC were studied for serum p53-Abs by enzyme-linked immunoabsorbent assay before and after treatment. The clinicopathologic features of p53 seropositive and p53 negative patients were compared. The relation between the presence of serum p53-Abs and p53 immunoreactivity of the resected specimens was examined. Three tumor markers (squamous cell carcinoma antigen [SCC-Ag], CYFRA21-1, and carcinoembryonic antigen [CEA]) were assessed to compare their sensitivities with serum p53-Abs. RESULTS: Fourteen of 35 patients (40%) were p53 seropositive. Relatively few patients tested positive for the other tumor markers: CEA, 11.4%; SCC-Ag, 14.3%; CYFRA21-1, 5.7%. There were no significant correlations between clinicopathologic features and p53 seropositivity except for tumor location. A strong correlation between p53 immunostaining and the presence of serum p53-Abs was observed (P = 0.003). Of the 14 patients with seropositive results, 12 turned seronegative after resection, and the other 2 experienced disease recurrence. CONCLUSIONS: Surveillance of serum p53-Abs is superior to the three tumor markers for detecting SESCC. This serum marker is also useful for the detection of p53 protein overexpression and for the monitoring of residual tumor cells. PMID- 11042561 TI - p27(Kip1) loss does not predict survival in patients with advanced gastric carcinoma. AB - BACKGROUND: p27(Kip1) is a cyclin-dependent kinase inhibitor whose loss is associated with disease progression and an unfavorable outcome in several malignancies. The authors studied its expression in a consecutive series of resected gastric carcinomas. METHODS: Expression of p27(Kip1) in 71 advanced gastric carcinomas and 10 lymph nodes containing metastases was determined using an avidin-biotin-peroxidase immunohistochemical method. The relations between p27(Kip1) expression and pathologic features, patient characteristics, and survival were analyzed. RESULTS: p27(Kip1) levels in gastric carcinomas ranged from 0.63-82.97% (median, 23. 10%; mean, 27.99%). There was no association found between p27(Kip1) expression and patient gender (P = 0.21), patient age (P = 0.13), tumor stage (P = 0.17), tumor grade (P = 0.22), or histologic type (P = 0.72). Univariate analysis showed that long term survival was related to stage (P < 0.0001) and grade (P = 0.03). However, tumors with p27(Kip1) levels above and below the median value were associated with a similar outcome, regardless of whether all cases (P = 0.19) or those without metastatic disease (P = 0.50) or those with residual or metastatic disease (P = 0.92) were included. When entered into a multivariate analysis, stage (P < 0.0001) and grade (P = 0.05), but not p27(Kip1) levels (P = 0.16), were found to be related to patient outcome. In lymph node metastases, p27(Kip1) expression (median, 16.5%) was similar to that found in the corresponding primary lesion (median, 30.9%). CONCLUSIONS: p27(Kip1) may play a role in the pathogenesis and progression of gastric carcinoma, but its expression is unlikely to be useful as a prognostic indicator, at least in European patients with advanced disease. PMID- 11042562 TI - Radioimmunoguided surgery for recurrent colorectal cancer manifested by isolated CEA elevation. AB - BACKGROUND: Carcinoembryonic antigen (CEA) is a sensitive marker for detecting recurrent colorectal carcinoma. An asymptomatic rise of CEA can precede by several months the detection of recurrent cancer by standard imaging modalities. Yet, surgeons are hesitant to operate solely on the basis of an observed increase in CEA. We investigated the ability of radioimmunoguided surgery to enhance the surgeon's capability of detecting intraabdominal disease in these patients. METHODS: Nineteen patients who underwent radioimmunoguided surgery for suspected tumor recurrence based solely on elevated CEA were included in the study. They underwent colonoscopy and CT of the abdomen and chest, all of which were negative. They then underwent scintigraphy scan with an anti-CEA monoclonal antibody (MoAb) labeled with (99m)Tc or Indium I-111. All patients were injected with the CC49 MoAb (an anti-TAG-72 tumor-associated glycoprotein) labeled with (125)I three weeks before surgery. During surgery, traditional exploration was followed by survey with a gamma-detecting probe. RESULTS: Traditional surgical exploration identified 26 recurrent tumors: 7 hepatic, 8 pelvic, 6 retroperitoneal, 3 colonic, 1 splenic, and 1 anastomotic. Radioimmunoguided surgical exploration confirmed all recurrent tumors and identified additional tumor sites in seven patients that resulted in changing the surgical plan. CEA scans correlated with intraabdominal findings in seven patients. Abdominal pathology did not correlate completely with the scans in three patients, and CEA scan results were undetermined in two patients. CONCLUSION: Patients with elevated CEA and no other findings should be operated upon without delay, and radioimmunoguided surgery should be used to enhance the surgeon's knowledge of the extent of disease. PMID- 11042563 TI - A North Central Cancer Treatment Group Phase II trial of 9-aminocamptothecin in previously untreated patients with measurable metastatic colorectal carcinoma. AB - BACKGROUND: Topoisomerase I inhibitors have demonstrated clinical activity in patients with metastatic colorectal carcinoma. The authors performed a Phase II study to evaluate the objective tumor response rate of 2 different doses and schedules of 9-aminocamptothecin (9-AC) in previously untreated patients with measurable recurrent metastatic colorectal carcinoma. METHODS: Fifty-one patients were registered. One schedule evaluated 9-AC given at 1100 microgram/m(2)/24 hours by continuous infusion for 72 hours along with granulocyte-colony stimulating factor at 5 microgram/kg/day on Days 5 through 12. Another schedule involved 9-AC at 480 microgram/m(2)/24 hours by continuous infusion for 120 hours on Days 1, 8, and 15 given every 4 weeks. RESULTS: Forty-eight of 51 patients (94%) were evaluable (28 patients who received 72-hour infusion and 20 patients who received 120-hour infusion) for response and toxicity. Significant hematologic toxicities were encountered, especially with the 72-hour infusion schedule, in which 43% (12 of 28) and 28% (8 of 28) experienced Grade 4 (National Cancer Institute Common Toxicity Criteria) leukopenia and thrombocytopenia, respectively. Grade 4 neutropenia was encountered in 61% (17 of 28) and 11% (2 of 19) of patients on the 72-hour and 120-hour infusion schedules, respectively. Diarrhea, nausea, vomiting, and hepatotoxicity were troublesome nonhematologic toxicities. Seventy-nine percent (11 of 14) and 57% (4 of 7) of the patients experiencing Grade 3 or 4 nonhematologic toxicity were on the 72-hour infusion schedule. Three patients died of chemotherapy-related toxicity. One response was observed in 48 evaluable patients (2%). CONCLUSIONS: 9-AC did not demonstrate sufficient antitumor activity and had unacceptable toxicity in previously untreated patients with metastatic colorectal carcinoma. PMID- 11042564 TI - Phase II trial of gemcitabine and UFT modulated by leucovorin in patients with advanced pancreatic carcinoma. The ONCOPAZ Cooperative Group. AB - BACKGROUND: Use of chemotherapy for advanced pancreatic carcinoma (APC) pursues a palliative objective. Gemcitabine is active against this tumor and shows in vitro synergism with 5-fluorouracil. UFT is a combination of tegafur (a prodrug of 5 flouorouracil) and uracil that can be given orally. The administration of UFT for several weeks may simulate the effects of a continuous infusion of 5 fluorouracil. The objective of the current study was to assess the efficacy and toxicity of the combination gemcitabine-UFT-leucovorin in the treatment of APC. METHODS: Forty-two patients with bidimensionally measurable APC were included. The study regimen consisted of gemcitabine 1000 mg/m(2) once weekly for 3 consecutive weeks, followed by a 1-week rest, intravenous 6S-steroisomer of leucovorin (6SLV) 250 mg/m(2) in 2 hours on Day 1, oral 6SLV 7.5 mg/12 hours on Days 2-14, and oral UFT 390 mg/m(2)/day (in 2 doses) on Days 1-14. Cycles were repeated every 4 weeks for a minimum of 3 per patient unless progressive disease was detected. RESULTS: One hundred eighty-three courses were given, with a median of 4 per patient. World Health Organization Grade 3-4 toxicity was: diarrhea in 7 patients (17%), leucopenia in 2 (5%), nausea/vomiting in 2 (5%), and anemia in 1 (4%). Among 38 patients evaluable for response, 6 achieved a partial response (16%; 95% confidence interval (CI), 6-31. 4), 15 had stable disease (39%), and 17 had progression (45%). Improvement in performance status and symptoms (pain, analgesic consumption, and weight) was present in 11 (29%) and 17 (45%) patients, respectively. Eighteen patients (47%; 95% CI, 31.5-54.5) experienced a clinical benefit response. CONCLUSIONS: The combination of gemcitabine-UFT-6SLV is convenient and moderately active and shows a low toxicity for the palliative treatment of patients with APC. PMID- 11042565 TI - A randomized study comparing two different schedules of administration of cisplatin in combination with gemcitabine in advanced nonsmall cell lung carcinoma. AB - BACKGROUND: This randomized trial was designed to investigate the feasibility, toxicity, and activity of two different schedules of gemcitabine plus cisplatin in previously untreated patients with advanced (International Union Against Cancer (UICC) Stage IIIB-IV) nonsmall cell lung carcinoma (NSCLC). METHODS: From February 1997 to September 1998, 82 patients with advanced NSCLC were entered onto the study and were randomized to gemcitabine 1000 mg/m(2) on Days 1, 8, and 15 plus cisplatin 80 mg/m(2) on Day 2 (arm A) or Day 15 (arm B) every 28 days. RESULTS: All the patients were assessable for toxicity (arm A/arm B: 151/177 cycles; median, 4 of 5 cycles per patient), and the following Grade 3-4 toxicities were reported (percentage of cycles in arm A vs. arm B): anemia, 7.9% and 2.3% (P < 0.05); leukopenia, 6.0% and 6.7%; thrombocytopenia, 15.0% and 1.6% (P < 0.01); no World Health Organization (WHO) Grade 3-4 nonhematologic toxicities were observed. These side effects led to gemcitabine dose reductions in 35.1% of courses in arm A and 22.0% of courses in arm B (P < 0.05) and to gemcitabine omissions in 28.5% of courses in arm A versus 7.3% of courses in arm B (P < 0.01). Dose intensities (DIs) of gemcitabine were 607.5 mg/m(2)/week in arm A and 711.6 mg/m(2)/week in arm B (P < 0.01); DIs of cisplatin were 18. 1 mg/m(2)/week in arm A and 18.8 mg/m(2)/week in arm B. The total delivered doses of gemcitabine were 9315.5 mg/m(2) in arm A and 12, 631.0 mg/m(2) in arm B (P < 0.01); the total delivered doses of cisplatin were 277.1 mg/m(2) in arm A and 333.0 mg/m(2) in arm B (P < 0.01). Response rates according to intention to treat were 40.4% (95% confidence interval [CI], 25.5-55.3) in arm A and 45% (95% CI, 29.5-60.5) in arm B. The overall median duration of response was 7.4 months; the median time to disease progression was 6 months (95% CI, 3-9) in arm A and 9 months (95% CI, 4-14) in arm B (P < 0.02); the median overall survival was 10 months (95% CI, 7.0-12.5) in arm A and 17 months (95% CI, 13.0-21.6) in arm B (P < 0.01); the 1-year survival rates were 34% and 63%, respectively. CONCLUSIONS: Our data show that arm B (cisplatin on Day 15) is less toxic than arm A (cisplatin on Day 2) and allows the administration of significantly higher total doses and dose intensities of chemotherapy. No significant differences in response rates were observed between the two schedules; patients on arm B experienced a significantly more prolonged progression free and overall survival; however, the study was not powered to detect differences in these outcomes. PMID- 11042566 TI - Frozen section analysis of sentinel lymph nodes in melanoma patients. AB - BACKGROUND: The sentinel lymph node biopsy (SLNB) is a diagnostic or staging option in the treatment of patients with cutaneous malignant melanoma (CMM) and is investigated intensively. A positive SLNB has appeared to identify patients who might have benefited from a lymph node dissection (LND). Intraoperative frozen section analysis (FSA) of the sentinel lymph node (SLN) during surgery would allow SLNB and LND to be performed in the same procedure. In the current study, we tested the reliability of FSA on the sentinel lymph node in patients with CMM. METHODS: Before definitive treatment of their melanomas began, FSA was performed on the SLNBs of 58 patients, whose median age was 56 (22-81) years, and who were 55% male and 45% female. Serial sections (500 micrometer interval), stained with routine hematoxylin and eosin and immunohistochemistry (S-100 and HMB-45), obtained definitive histology of the sentinel lymph node. RESULTS: Detection of the sentinel lymph node was possible in 56 patients (97%). Sixty-one SLNBs were performed in these patients. FSA detected metastases in 5 of 108 SLN (5%) in 5 patients. This was upgraded after definitive histology to 13 SLN (12%) in 11 patients (20%). Sensitivity of the FSA was 38%. After a median follow-up of 35 (range: 24-54) months, the false-negative rate of the SLN was 4% (2 patients). CONCLUSION: The combination of the low sensitivity of FSA and a finding that only 12% of the SLNBs contained metastases does not justify routine use of FSA on the SLN of patients with CMM. PMID- 11042567 TI - Routine bone scintigraphy in primary staging of soft tissue sarcoma; Is it worthwhile? AB - BACKGROUND: The incidence of bone metastases in soft tissue sarcoma (STS) patients seems to be low but has not been studied separately. In this study, the authors aimed to determine the value of routine radionuclide bone scanning in preoperative staging of STS patients. METHODS: Preoperative bone scans were evaluated retrospectively in 109 consecutive patients (median age, 44 years; range, 1-86) with intermediate or high grade STS. Scans were scored in 3 categories: 1, metastases very likely; 2, equivocal; and 3, normal or benign lesions. RESULTS: Category 1 scans were found in 8 of 109 patients (7%); in all 8 patients, bone metastases were confirmed. Six of these eight patients reported pain, and all had additional lung, bone marrow, or lymph node metastases. The highest rate (17%) was found in the rhabdomyosarcoma subgroup (n = 18). Category 2 (equivocal) scans were present in 12 of 109 patients (11%), in all of which bone metastases were excluded through additional investigations. Category 3 (normal) scans were found in 81%. Bone metastases were at least as frequent as lung metastases (4%) and were the single site of systemic disease in 4%. The rate of bone metastases was 55% in patients with bone pain versus 2% in patients without pain. CONCLUSIONS: Bone metastases in primary STS patients are rare (7%) yet in this study at least as frequent as lung metastases. The low rate in asymptomatic patients versus the high rate in symptomatic patients supports the use of bone scanning in symptomatic patients only. The yield of routine bone scanning is low. PMID- 11042568 TI - Quantitative analysis of estrogen receptor-alpha and -beta messenger RNA expression in breast carcinoma by real-time polymerase chain reaction. AB - BACKGROUND: Estrogen action is mediated not only through a classic estrogen receptor (ER) (ER-alpha) but also through a second ER (ER-beta) that has a structure and function similar to ER-alpha. A correlation between ER-beta mRNA expression with ER and progesterone receptor (PR) protein levels as well as prognostic factors remains to be established in breast carcinoma. METHODS: The authors conducted a quantitative analysis of ER-alpha and ER-beta mRNA expression in 116 breast tumors using real-time polymerase chain reaction (PCR), and investigated a possible correlation between ER-alpha and ER-beta mRNA expression and ER and PR status as determined by enzyme immunoassay as well as with various prognostic factors. RESULTS: ER-alpha mRNA levels were significantly (P < 0.01) higher in ER positive compared with ER negative tumors. Conversely, ER-beta mRNA levels were significantly (P < 0.01) lower in ER positive compared with ER negative tumors. Accordingly, the ratio of ER-beta to ER-alpha was significantly (P < 0.01) higher in ER negative compared with ER positive tumors. A subset analysis based on ER and PR status showed that ER-beta mRNA levels as well as the ratios of ER-beta to ER-alpha mRNA level were highest in ER negative and PR negative tumors (P < 0.05). ER-alpha mRNA levels were significantly (P < 0.05) higher in postmenopausal compared with premenopausal tumors. Histologic Grade 3 tumors showed a significant decrease in ER-alpha mRNA levels compared with Grade 1 and 2 tumors (P < 0.01 and P < 0.05, respectively). No significant correlation between ER-alpha and ER-beta mRNA levels and histologic type, tumor size, or lymph node status was observed. CONCLUSIONS: An absolute and relative increase in ER-beta mRNA levels in ER negative and PR negative breast tumors, which rarely respond to endocrine therapy, suggests the possible involvement of up-regulation of ER-beta mRNA in the development of estrogen-independent tumors. PMID- 11042569 TI - The effect of less than definitive care on breast carcinoma recurrence and mortality. AB - BACKGROUND: Risk factors for breast carcinoma offer few opportunities for prevention; thus, the reduction of morbidity and mortality among breast carcinoma patients must remain a priority. The objective of this study was to measure the effects of less than definitive care for patients with breast carcinoma on disease recurrence and mortality. METHODS: The prognostic evaluation and treatment received by an inception cohort of 494 women was characterized. Three hundred ninety women ages 45-90 years with local or regional breast carcinoma who were diagnosed between 1984 and 1986 and were treated at one of eight Rhode Island hospitals comprised the final cohort. Disease recurrence and mortality were ascertained through December 31, 1996. Candidate determinants of outcomes were a less than definitive prognostic evaluation and less than definitive primary therapy-adjusted for confounding by patient age, extent of disease, and comorbid diseases. RESULTS: During the first 5 years of follow-up, patients who received a less than definitive prognostic evaluation had an adjusted relative hazard of recurrence of 1.7 (95% confidence interval, 1.0-2.7) and an adjusted relative hazard for breast carcinoma mortality of 2.2 (95% confidence interval, 1.2-3.9). Patients who received less than definitive therapy had an adjusted relative hazard of recurrence of 1.6 (95% confidence interval, 1.0-2.5), and an adjusted relative hazard of breast carcinoma mortality of 1.7 (95% confidence interval, 1.0-2.8). CONCLUSIONS: Breast carcinoma patients who receive less than definitive care are at excess risk for disease recurrence and mortality. Women with early stage breast carcinoma should be treated in accordance with existing guidelines. PMID- 11042570 TI - Prognostic value of DNA cytometry in 281 premenopausal patients with lymph node negative breast carcinoma randomized in a control trial: multivariate analysis with Ki-67 index, mitotic count, and microvessel density. AB - BACKGROUND: The clinical relevance of DNA image cytometry (ICM) and flow cytometry (FCM) remains under investigation in breast carcinoma. The objective of the current work was to study the prognostic value of DNA ICM and FCM in a series of patients randomized in a control trial. A multivariate analysis has been performed including other factors still under investigation such as Ki-67 index, mitotic count, microvessel density, and P53 and Bcl-2 expression. METHODS: Two hundred and eighty-one patients were randomized in the European Organization for Research and Treatment of Cancer 10854 trial comparing surgery followed by one course of perioperative chemotherapy versus surgery alone. Tumor parameters studied were pT, multicentricity, tumor grading according to modified Scarff Bloom-Richardson, estrogen receptors, mitotic count per 1.7 mm(2), MIB-1, and BCL 2 scores, microvessel density, and p53 expression. ICM DNA parameters studied from paraffin embedded specimens, were DNA ploidy, proliferative index, 2c deviation index, malignancy grade, and Auer-Baldetorp typing. FCM DNA parameters analyzed on the same samples were ploidy and S-phase fraction statistics. The influence of tumor parameters, and DNA parameters on overall survival (OS), disease free survival (DFS), and metastasis-free survival (MFS) was evaluated using the Cox model. Median follow-up was 82 months. RESULTS: For OS, the prognostic parameters retained were pathologic tumor size (pT) and mitotic index (MI). Overall survival was 94% and 68% for tumors pT1/MI less than 10 and pT2-3 MI greater than or equal to 10, respectively. For DFS, age, multicentricity, and grading according to modified Scarff and Bloom were predicting factors with the same relative risk. Disease free survival was 96%, 78% and 68% respectively, when 1, 2, or 3 of those factors were present. For MFS, the only retained predicting factor was MI. MFS was 97% and 73% when MI was less than 10 and MI was greater than or equal to 10, respectively. CONCLUSIONS: Evaluation of proliferative compartment was the most important predicting factor for OS and MFS in the current series of premenopausal lymph node negative patients with breast invasive carcinoma. When working on paraffin embedded tissue, the best way of assessing it was MI count. ICM DNA analysis results were not selected in multivariate analysis. DNA analysis by FCM should be considered as an unsuitable technique when working on paraffin embedded tissue. PMID- 11042571 TI - Clinical significance of microsatellite instability in endometrial carcinoma. AB - BACKGROUND: The purpose of this study was to compare the clinical characteristics of endometrial carcinomas with and without microsatellite instability (MSI). METHODS: The authors prospectively acquired DNA from patients with endometrial carcinomas at Washington University Medical Center. Tumors were assigned MSI (+) status when two or more of five microsatellite repeat markers revealed novel bands in tumor DNA not present in the corresponding normal DNA. Clinical characteristics and survival data of patients with and without MSI were abstracted from patient charts. Statistical significance was calculated with the chi-square test, and survival was assessed with Kaplan-Meier methods. RESULTS: The authors found 65 of 70 (93%) patients with MSI (+) tumors to be of white race, whereas only 124 of 159 (78%) patients with MSI (-) tumors were white (P = 0.012). Advanced disease (International Federation of Gynecology and Obstetrics Stage III-IV) was observed in 9 of 70 (13%) MSI (+) patients and 44 of 159 (28%) MSI (-) patients (P = 0.017). In addition, aggressive histologic subtypes were observed less frequently in MSI (+) tumors (6/70 [8%]) than in MSI (-) tumors (30 of 159 [19%]) (P = 0.034). Race and stage were shown by multivariate analysis to be different in MSI (+) and MSI (-) patients. Recurrence and overall survival were similar in the two groups. CONCLUSIONS: Patients with MSI (+) tumors were more likely to be of white race and to present with early stage disease. Further investigation is needed to explain why patients with MSI (+) tumors have similar survival to patients with MSI (-) tumors, despite presenting at earlier stages, being of white race, and being less likely to be associated with virulent histologic subtypes. PMID- 11042572 TI - The accuracy of endometrial sampling in the diagnosis of patients with endometrial carcinoma and hyperplasia: a meta-analysis. AB - BACKGROUND: Endometrial assessment by means of biopsy or sampling of endometrial cells is a minimally invasive alternative for dilatation and curettage (D&C) or hysteroscopy. The use of this technique is believed to reduce the cost of the diagnostic work-up for abnormal uterine bleeding without reducing accuracy. Because the authors were not aware of any systematic review of this test, they performed a meta-analysis to assess the accuracy of endometrial sampling devices in the detection of endometrial carcinoma and atypical hyperplasia. METHODS: The authors searched the literature for studies published between 1966 and 1999 comparing the results of endometrial sampling with findings at D&C, hysteroscopy, and/or hysterectomy. They found 39 studies that included 7914 women. For each study, the fraction of patients was calculated in which endometrial sampling failed. Furthermore, the authors calculated the fraction of cases of endometrial carcinoma and atypical hyperplasia that were identified correctly as well as the fraction of women in whom these diseases were diagnosed false positively. RESULTS: The detection rate for endometrial carcinoma was higher in postmenopausal women compared with premenopausal women. In both postmenopausal and premenopausal women, the Pipelle was the best device, with detection rates of 99. 6% and 91%, respectively. For the detection of atypical hyperplasia, there was only one study that reported explicitly on postmenopausal women, thereby hampering the possibility of subgroup analysis. Again, the Pipelle was the most sensitive technique with a sensitivity of 81%. The specificity of all devices was > 98%. CONCLUSIONS: Endometrial biopsy with the Pipelle is superior to other endometrial techniques in the detection of endometrial carcinoma and atypical hyperplasia. The accuracy of the Pipelle is higher in postmenopausal women compared with premenopausal women. PMID- 11042573 TI - Microsatellite analysis at 10q25-q26 in Sardinian patients with sporadic endometrial carcinoma: identification of specification patterns of genetic alteration. AB - BACKGROUND: Loss of heterozygosity (LOH) at chromosome 10q25-q26 has been reported previously in endometrial carcinoma (EC), suggesting the presence of tumor suppressor gene(s). Nevertheless, frequency of genome-wide microsatellite instability (MSI) has been demonstrated higher in EC than in other common malignancy, mostly due to defective DNA mismatch repair. The authors further evaluated the role of the chromosome 10q25-q26 in endometrial tumorigenesis as well as the clinical significance of any observed genetic alteration in sporadic EC. METHODS: Paired normal and tumor samples from 94 Sardinian patients with sporadic EC at various stages of disease were screened by polymerase chain reaction (PCR)-based microsatellite analysis. Genomic DNA was isolated from paraffin embedded tissues and amplified by PCR using microsatellite markers spanning approximately 14 cM at 10q25-q26. Microsatellite instability was studied at four loci mapping to different chromosomal locations. RESULTS: Thirty-two (34%) EC patients were found negative for genetic alterations within the 10q25 q26 region. Among the remaining 62 (66%) EC cases, the authors identified 1) a minimum consensus region of LOH of approximately 1 cM, between D10S610 and D10S542 markers; and 2) a subset of tumors with prevalence of instability at 10q25-q26 (10qMI+), as expression of the presence of a MSI+ phenotype. CONCLUSIONS: The authors' data establish the existence of significant correlations between disease stages and 10qMI+ (with or without MSI+). However, longer follow-up and additional studies are required to define the clinical significance of these findings as prognostic factors. Moreover, the minimum region of LOH at 10q25-q26 will be further analyzed for identifying the putative tumor suppressor gene involved in EC pathogenesis. PMID- 11042574 TI - Steroid hormone receptors and long term survival in invasive ovarian cancer. AB - BACKGROUND: Steroid hormone receptors are important determinants of prognosis and predictive behavior in tumor tissues of several origins. Since their role in ovarian cancer is still controversial, we investigated the prevalence and prognostic impact of the estrogen (ER) and progesterone (PR) receptors and combinations (ER+PR+, ER+PR-, ER-PR+, and ER-PR-) in a comparably large number of patients with a long clinical follow-up. METHODS: The present analysis included 186 patients with invasive ovarian carcinomas treated at the Department of Obstetrics and Gynecology of the Justus-Liebig-University Giessen between 1982 and 1996, the follow-up lasting up to 15.8 years (median 2.4 yrs). The expression of ER and PR was assessed by immunohistochemistry using alkaline phosphatase antialkaline phosphatase in microwave pretreated, formalin fixed, and paraffin embedded specimens of the primary tumors and was evaluated semiquantitatively using a standardized immunoreactive scoring system. Receptor expression and combinations were compared to clinical, histologic and prognostic factors, the tumor proliferation, and the clinical outcome. RESULTS: Kaplan-Meier survival analyses supported the favorable prognostic value of PR and its level of expression in ovarian carcinomas. Especially the ER-PR+ combination, which accounted for 10.2% of all tumors, showed a significantly superior prognosis when compared with all other combinations (survivors 15 of 19 vs. 67 of 167, log rank P = 0.009) and was associated with early stage, low ascites quantity, and higher tumor differentiation. Five-year survival rates were 13/16 (81.3%) for ER-PR+ tumors versus 58/128 (45.3%) for all other steroid hormone receptor combinations. Residual analysis proved the results. CONCLUSIONS: The determination of steroid hormone receptor status offers additional prognostic information in ovarian carcinomas. Especially the ER-PR+ phenotype predicts a favorable tumor biology and long term survival, probably reflecting functional effects on tumor proliferation, differentiation, and cellular apoptosis. PMID- 11042575 TI - A nationwide charge comparison of the principal treatments for early stage prostate carcinoma. AB - BACKGROUND: The diagnosis and treatment of men with early stage prostate carcinoma is expensive and controversial, yet the similarities in reported survival rates has underscored the importance of ascertaining the relative charges of different forms of therapy. METHODS: Patient specific data on demographic characteristics, hospital and physician resource use, and charges were obtained from the Health Care Financing Administration for 1993 through 1996. The inpatient, outpatient, and part B claims from men with a new diagnosis of prostate carcinoma were captured from the quarter of the year in which biopsy was performed through the two quarters after treatment. Charges are reported in inflation-unadjusted dollars. RESULTS: Of 10,107 men treated for early stage prostate carcinoma, 58% received external beam radiation therapy (XRT), 35% had radical prostatectomy, and 7% underwent brachytherapy. Over the 4 years, use of XRT decreased 19% whereas use of brachytherapy increased 21%. Men aged 65-69 years were more likely to have radical prostatectomy, but after age 70 years, XRT predominated. The most expensive treatments were radical prostatectomy with adjuvant XRT ($31,329) and brachytherapy with pretreatment XRT ($24,407). Cost of radical prostatectomy alone was more than XRT alone ($19,019 vs. 15,937; P < 0.05) or brachytherapy alone ($15,301; P < 0.05). Treatment utilization varied with age, race, and geographic region. CONCLUSIONS: The mean charges for the workup, treatment, and 6 month follow-up of patients treated for early stage prostate carcinoma ranged between $15,301 and $31,329, with significant treatment group differences. Without a clear survival advantage from one form of treatment, issues such as costs, quality of life, and patient preferences take on paramount importance. PMID- 11042576 TI - Detailed mapping of prostate carcinoma foci: biopsy strategy implications. AB - BACKGROUND: Prostate carcinoma exhibits considerable anatomic heterogeneity. Detailed characterization of prostate carcinoma distribution could lead to improved detection procedures and biopsy strategies. We mapped all 607 tumor foci from 180 serially sectioned whole mount radical prostatectomy specimens and used a computer algorithm to plot and summarize the distribution of these foci. We investigated whether specimen and clinical variables predicted differences in tumor distribution. METHODS: The volume and anatomic location of each tumor focus were determined and digitized. A computer-based algorithm was used to fit the digitized tumor foci to a paradigm prostate. Pseudo-color summary plots of tumor distribution then were computed for selected cases. RESULTS: Of the 180 specimens, 149 (83%) specimens had more than one cancer focus. Most foci (448 of 607 tumor foci, 74%) were in the peripheral zone (PZ). PZ foci near the apex had a significant midline component. Toward the base, PZ foci diverged laterally. Only 3 (2%) of 180 specimens contained foci solely in the transition zone (TZ). Total TZ cancer volume was 50% decrease in serum prostate-specific antigen (PSA) in 6 of 25 patients (24% of patients; 95% confidence interval (CI) 9-45%). The median duration of PSA response was 10 weeks (range: 3-39 weeks). Of the five men with bidimensionally measurable disease, none achieved a complete or partial response. There were no documented improvements in post-treatment bone scans. Median overall survival time was 14.1 months. CONCLUSIONS: The combination of vinorelbine and estramustine is a well-tolerated and modestly active regimen in men with androgen-independent metastatic prostate cancer. PMID- 11042580 TI - The effect of disease and treatment-related factors on biopsy results after prostate brachytherapy: implications for treatment optimization. AB - BACKGROUND: Posttreatment prostate biopsy is a method of assessing local control after irradiation for prostate carcinoma. An analysis of the effect of disease- and treatment-related factors on biopsy results after prostate brachytherapy was performed to aid in patient selection and treatment optimization. METHODS: Two hundred sixty-eight patients underwent posttreatment prostate biopsy (6-8 cores) 2 years after brachytherapy alone without external beam irradiation. Follow-up ranged from 24 to 111 months (median, 43 months). Implants were performed using a real-time ultrasound guided technique with the isotopes (125)I in 186 and (103)Pd in 82 patients. Ninety-eight patients underwent hormonal therapy (HT) 3 months before and 2-3 months after implant. Implant dose was defined as the D90 (dose delivered to 90% of the gland from the dose volume histogram generated using 1 month computed tomography-based dosimetry). RESULTS: Overall, 89% of patients (238 of 268) had negative biopsies. A positive biopsy was a predictor of biochemical failure. Patients with a positive biopsy had a 5-year freedom from biochemical failure of 40% versus 76% for patients with a negative biopsy (P = 0.0003). Univariate and multivariate analysis found that risk group, HT, and implant dose significantly affected biopsy outcome. Patients with low risk features (prostate specific antigen [PSA] 10 ng/mL or Gleason score >/= 7 or classification T2b or higher) (n = 164) (P = 0.008). Hormonal therapy was associated with a negative biopsy rate of 98% versus 84% for implant alone (P = 0.003). Patients receiving a high implant dose (D90 >/= 140 grays [Gy] for (125)I or >/= 100 Gy for (103)Pd) (n = 174) had a negative biopsy rate of 95% versus 77% for those receiving a low dose (D90 < 140 Gy for (125)I or < 100 Gy for (103)Pd) (n = 87; P < 0.001). CONCLUSIONS: Biopsy results support the use of brachytherapy without external beam irradiation for patients with low risk features and highlight the importance of achieving an adequate implant dose. PMID- 11042581 TI - Reduction of tumor burden and stabilization of disease by systemic therapy with anti-CD20 antibody (rituximab) in patients with primary cutaneous B-cell lymphoma. AB - BACKGROUND: Primary cutaneous B-cell lymphoma (CBCL) is characterized by restriction to the skin, a high incidence of recurrence after various treatment modalities, and a variable but mostly favorable prognosis. METHODS: Ten patients with long standing primary CBCL (3 with follicular CBCL, 5 with cutaneous, large B-cell lymphoma, 1 with diffuse large cell lymphoma, and 1 with extranodal large cell lymphoma) were treated by intravenous application of a chimeric antibody against the CD20 transmembrane antigen that is present on malignant and normal B cells. In 6 of 10 patients, several treatment attempts either had failed or could not be used due to severe side effects or underlying disease. RESULTS: The treatment regimen resulted in two complete regressions, five partial responses, and one mixed response, and two patients did not respond to the treatment. No severe side effects occurred, except for slight pain in the nodules after infusion and an urticarial reaction at the tumor sites. A prolonged, complete disappearance of B-cells from the peripheral blood was observed. The immunoglobulin serum levels and inflammatory markers were unchanged. Histologic examination of biopsies from two regressing tumor nodes showed necrotic tumor cells and infiltration with CD8 positive cells. CONCLUSIONS: Intravenous therapy with the anti-CD20 antibody rituximab is a nontoxic and effective treatment for patients with primary cutaneous B-cell lymphoma. PMID- 11042582 TI - Hepatoblastoma presenting with lung metastases: treatment results of the first cooperative, prospective study of the International Society of Paediatric Oncology on childhood liver tumors. AB - BACKGROUND: The prognosis of children who are affected by hepatoblastoma (HB) that presents with lung metastases has always been considered very poor. In light of the overall improvement in the survival of HB patients since the introduction of cisplatin (CDDP) in the therapeutic armament of this tumor, the question has been raised whether patients with metastatic HB also would benefit from this drug. The purpose of the current study was to address this issue by analyzing the treatment outcome of those patients presenting with metastases who entered into the first HB study on childhood liver tumors conducted by the International Society of Paediatric Oncology (SIOPEL 1). METHODS: SIOPEL 1 was a prospective, international, multicentric, single-arm study based on preoperative chemotherapy that was open to patient registration from January 1990 to February 1994. After undergoing a biopsy, patients received four courses of CDDP (80 mg/m(2) in a 24 hour, continuous infusion) on Day 1 followed by doxorubicin (60 mg/m(2) in a 48 hour, continuous infusion) on Days 2 and 3 (PLADO). Surgery was performed after four courses of PLADO and was followed by two more courses. Untreated children age < 16 years with biopsy-proven HB were eligible for the study. Metastatic spread was assessed by chest X-ray and, where available, lung computed tomography scan. RESULTS: Thirty-one of 154 children that entered into the trial presented with metastases. Eight children presently are alive with no evidence of disease (NED) after being treated with protocol therapy only (median follow-up, 60 months); nine children are alive with NED after having failed PLADO and having been rescued with alternative therapies (median follow-up, 80 months). The 5-year overall and event free survival rates for these children were 57% (95% confidence interval, 39-75%) and 28% (95% confidence interval, 12-44%), respectively. Persistent lung disease was the main reason for PLADO failure (17 of 23 patients; 74%). CONCLUSIONS: The SIOPEL 1 therapeutic strategy seems to cure 25% of the HB patients who present with metastases. However, further chemotherapy and the use of thoracotomies still can save significant numbers of these children. PMID- 11042584 TI - Merkel cell carcinoma: Review of 22 cases with surgical, pathologic, and therapeutic considerations. PMID- 11042583 TI - Living with cancer: "good" days and "bad" days--what produces them? Can the McGill quality of life questionnaire distinguish between them? AB - BACKGROUND: To determine the impact of care on quality of life (QOL), or to detect a change in QOL over time, measures of QOL must remain stable when QOL is stable (test-retest reliability) and change when QOL changes (responsiveness). This study addresses these issues for the McGill Quality of Life Questionnaire (MQOL). Unlike other studies that use disease status to indicate whether QOL has remained stable or changed, in this study the patient determines QOL stability or change. The authors also sought to clarify the determinants of "good" and "bad" days for oncology patients. METHODS: Patients attending an oncology outpatient clinic or who were being treated by a palliative care service were asked to complete MQOL 4 times: on days they judged to be "good," "average," and "bad" and 2 days after the first completion. They also were asked to directly rate the change in their QOL during the intervals between MQOL completion and to report the most important determinants of their good and bad days. RESULTS: The test retest reliability of MQOL as measured by an intraclass correlation coefficient ranged from 0.69 to 0.78. All MQOL scores were significantly different on good, average, and bad days, except for the support subscale, in both clinical settings. Five domains were determinants of QOL: physical symptoms, physical functioning, psychologic well-being, existential well-being, and relationships. CONCLUSIONS: MQOL's reliability and responsiveness suggest it can be used to determine changes in the QOL of groups. The results allow interpretation of changes in MQOL scores with respect to meaning of the change to oncology patients. This in turn is helpful to determine the sample size required in future studies. Some of the domains important to the QOL of oncology patients are not included in widely used measures of QOL. PMID- 11042585 TI - Author reply PMID- 11042586 TI - Hormone replacement therapy and lobular breast carcinoma. PMID- 11042587 TI - Author reply PMID- 11042588 TI - B2 bradykinin receptor immunoreactivity in rat brain. AB - Bradykinin has long been known to exist in the central nervous system and has been hypothesized to mediate specific functions. Despite an increasing understanding of the functions of bradykinin, little is known about the cell types expressing the bradykinin receptor within the brain. The present investigation employed a monoclonal antibody directed against the 15-amino-acid portion of the C-terminal of the human bradykinin B2 receptor to establish the cellular distribution of bradykinin B2 receptor immunoreactivity in the rat brain. Bradykinin B2 receptor immunoreactivity was ubiquitously and selectively observed in neurons, including those within the olfactory bulb, cerebral cortex, hippocampus, basal forebrain, basal ganglia, thalamus, hypothalamus, cerebellum, and brainstem nuclei. Bradykinin B2 receptor immunoreactivity was also present in the circumventricular organs including choroid plexus, subfornical organ, median eminence, and area postrema. Double-labeling experiments colocalizing the bradykinin B2 receptor with the neuronal marker NeuN or the astrocytic marker glial fibrillary acidic protein revealed that virtually 100% of the bradykinin B2 receptor-immunoreactive positive cells were neurons. The widespread distribution of bradykinin B2 receptor immunoreactivity in neuronal compartments suggests a greater than previously appreciated role for this peptide in neuronal function. PMID- 11042589 TI - Loss of nucleus basalis neurons containing trkA immunoreactivity in individuals with mild cognitive impairment and early Alzheimer's disease. AB - Recent studies indicate that there is a marked reduction in trkA-containing nucleus basalis neurons in end-stage Alzheimer's disease (AD). We used unbiased stereological counting procedures to determine whether these changes extend to individuals with mild cognitive impairment (MCI) without dementia from a cohort of people enrolled in the Religious Orders Study. Thirty people (average age 84.7 years) came to autopsy. All individuals were cognitively tested within 12 months of death (average MMSE 24.2). Clinically, 9 had no cognitive impairment (NCI), 12 were categorized with MCI, and 9 had probable AD The average number of trkA immunoreactive neurons in persons with NCI was 196, 632 +/- 12,093 (n = 9), for those with MCI it was 106,110 +/- 14,565, and for those with AD it was 86,978 +/- 12,141. Multiple comparisons showed that both those with MCI and those with AD had significant loss in the number of trkA-containing neurons compared to those with NCI (46% decrease for MCI, 56% for AD). An analysis of variance revealed that the total number of neurons containing trkA immunoreactivity was related to diagnostic classification (P < 0.001), with a significant reduction in AD and MCI compared to NCI but without a significant difference between MCI and AD. Cell density was similarly related to diagnostic classification (P < 0.001). There was a significant correlation with the Boston Naming Test and with a global score measure of cognitive function. The number of trkA-immunoreactive neurons was not correlated with MMSE, age at death, education, apolipoprotein E allele status, gender, or Braak score. These data indicate that alterations in the number of nucleus basalis neurons containing trkA immunoreactivity occurs early and are not accelerated from the transition from MCI to mild AD. PMID- 11042590 TI - Ultrastructural synaptic organization of axon terminals in the ventral part of the cat oral pontine reticular nucleus. AB - In an attempt to contribute to the current knowledge of the brainstem reticular formation synaptic organization, the ultrastructure and distribution of synaptic terminal profiles on neurons in the ventral part of the oral pontine reticular nucleus (vRPO), the rapid eye movement (REM) sleep-induction site, were studied quantitatively. Terminals with asymmetric contacts and rounded vesicles were classified according to vesicle density as type I or II (high or low density, respectively). The area, apposed perimeter length, and mitochondrial area of type I terminals, on average, were significantly smaller than those of type II terminals. Type III and IV terminals had symmetric contacts and oval and/or flattened vesicles; type III terminals formed synapses between them and on initial axons. Type V and VI terminals showed characteristics intermediate to those of asymmetric and symmetric synapses. Interestingly, some terminal features were related to both terminal area and postsynaptic dendritic diameter. The percentages of different synapses sampled on somata were as follows: asymmetric synapses (usually formed by type II terminals; mean +/- S.D.), 26.4% +/- 3%; symmetric synapses, 46.7% +/- 5.2%; and intermediate synapses, 26.9% +/- 6.1%. The percentages of different synapses sampled on dendrites were asymmetric synapses, 62.1% +/- 9%; symmetric synapses, 25.6% +/- 8.1%; and intermediate synapses, 12.3% +/- 1.7%. Comparison between large- and small-diameter dendrites revealed that the percentages of symmetric synapses and type II terminals decreased, whereas the percentages of type I terminals increased as postsynaptic dendritic diameters became smaller. Synaptic density was approximately four times lower on somata than on dendrites. The vRPO synaptic organization reflects some patterns that are similar to those found in other regions of the central nervous system as well as specific synaptic patterns that are probably related to its functions: the generation and maintenance of REM sleep and the control of eye movement or limb muscle tone. PMID- 11042591 TI - Absence of p75(NTR) expression reduces nerve growth factor immunolocalization in cholinergic septal neurons. AB - Septal axons provide a cholinergic innervation to the nerve growth factor (NGF) producing neurons of the mammalian hippocampus. These cholinergic septal afferents are capable of responding to target-derived NGF because they possess trkA and p75(NTR), the two transmembrane receptors that bind NGF and activate ligand-mediated intracellular signaling. To assess the relative importance of p75(NTR) expression for the responsiveness of cholinergic septal neurons to hippocampally derived NGF, we used three lines of mutant and/or transgenic mice: p75(-/-) mice (having two mutated alleles of the p75(NTR) gene), NGF/p75(+/+) mice (transgenic animals overexpressing NGF within central glial cells and having two normal alleles of the p75(NTR) gene), and NGF/p75(-/-) mice (NGF transgenic animals having two mutated alleles of the p75(NTR) gene). BALB/c and C57B1/6 mice (background strains for the mutant and transgenic lines of mice) were used as controls. Both lines of NGF transgenic mice possess elevated levels of NGF protein in the hippocampus and septal region, irrespective of p75(NTR) expression. BALB/c and C57Bl/6 mice display comparably lower levels of NGF protein in both tissues. Despite differing levels of NGF protein, the ratios of hippocampal to septal NGF levels are similar among BALB/c, C57B1/6, and NGF/p75(+/+) mice. Both p75(-/-) and NGF/p75(-/-) mice, on the other hand, have markedly elevated ratios of NGF protein between these two tissues. The lack of p75(NTR) expression also results in a pronounced absence of NGF immunoreactivity in cholinergic septal neurons of p75(-/-) and NGF/p75(-/-) mice. BALB/c, C57B1/6, and NGF/p75(+/+) mice, on the other hand, display NGF immunoreactivity that appears as discrete granules scattered through the cytoplasm of cholinergic septal neurons. Elevated levels of NGF in the hippocampus and septal region coincide with hypertrophy of cholinergic septal neurons of NGF/p75(+/+) mice but not of NGF/p75(-/-) mice. Levels of choline acetyltransferase (ChAT) enzyme activity are, however, elevated in the septal region and hippocampus of both NGF/p75(+/+) and NGF/p75(-/-) mice, compared with control mice. These data indicate that an absence of functional p75(NTR) expression disrupts the normal cellular immunolocalization of NGF by cholinergic septal neurons but does not affect the ability of these neurons to respond to elevated levels of NGF, as determined by ChAT activity. PMID- 11042592 TI - Expression of calcium-binding proteins in the diencephalon of the lizard Psammodromus algirus. AB - This work is a study of the distribution pattern of calbindin-D28k, calretinin, and parvalbumin in the diencephalic alar plate of a reptile, the lizard Psammodromus algirus, by using the prosomeric model (Puelles [1995] Brain Behav Evol 46:319-337), which divides the alar plate of the diencephalon into the caudorostrally arranged pretectum (p1), dorsal thalamus plus epithalamus (p2), and ventral thalamus (p3). Calbindin and calretinin are more extensively expressed in the dorsal thalamus than in the neighboring alar regions, and therefore these calcium-binding proteins are particularly suitable markers for delimiting the dorsal thalamus/epithalamus complex from the ventral thalamus and the pretectum. Conversely, parvalbumin is more intensely expressed in the pretectum and ventral thalamus than in the dorsal thalamus/epithalamus complex. Within the dorsal thalamus, calcium-binding protein immunoreactivity reveals a three-tiered division. The pretectum displays the most intense expression of parvalbumin within the diencephalon. Virtually all nuclei in the three sectors of the pretectum (commissural, juxtacommissural, and precommissural) present strong to moderate expression of parvalbumin. We compare the distribution of calcium binding proteins in the diencephalon of Psammodromus with other vertebrates, with mammals in particular, and suggest that the middle and ventral tiers of the reptilian dorsal thalamus may be comparable to nonspecific or plurimodal posterior/intralaminar thalamic nuclei in mammals, on the basis of the calcium binding protein expression patterns, as well as the hodological and embryological data in the literature. PMID- 11042593 TI - Immunolabeling reveals cellular localization of the N-methyl-D-aspartate receptor subunit NR2B in neurosecretory cells but not astrocytes of the rat magnocellular nuclei. AB - Previous studies suggest that activation of N-methyl-D-aspartate (NMDA) receptors facilitates phasic firing and spike clustering displayed by magnocellular neuroendocrine cells (MNCs) of the supraoptic (SON) and paraventricular nucleus of the hypothalamus (PVN). Osmotic stimulation produces similar activity patterns which, in turn, can lead to enhanced release of vasopressin and oxytocin from MNCs. Our laboratory has shown that dehydration regulates the expression of the NMDA receptor subunits, NR1 and NR2B, in the SON and PVN, suggesting their involvement in osmoregulation. In the present study, we examined the cellular localization of NR2B, one of the glutamate-binding subunits of the NMDA receptor, with an NR2B-specific antibody. Using double-label immunohistochemistry and three different detection methods with metallic, peroxidase, and fluorescence markers, it was found that both vasopressin and oxytocin-producing MNC populations synthesize NR2B. The incidence of NR2B colocalization with vasopressin neurophysin in the SON and lateral magnocellular PVN (PVL) was 0.95 and 0.91, respectively. For oxytocin-neurophysin, the corresponding values were 0.97 and 0.95, respectively. Furthermore, the extent of colocalization in MNCs of the SON, PVL, retrochiasmatic SON, and accessory neurosecretory nuclei was similar. Astrocytes associated with the SON, and identified with antibodies targeting glial fibrillary acidic protein (GFAP) or vimentin, were not colabeled with NR2B. Our results demonstrate that NR2B protein is expressed by almost all MNCs and that it is equally prevalent in vasopressinergic and oxytocinergic populations of various magnocellular neuroendocrine nuclei supporting a role of NMDA receptors in MNC-mediated neurosecretory processes. Although NR2B may form part of functional NMDA receptors on MNCs, it is probably not present on astrocytes associated with nearby MNCs. PMID- 11042594 TI - AMPA receptor subunit expression in trigeminal neurons during postnatal development. AB - Trigeminal motoneurons (Mo5) and mesencephalic trigeminal neurons (Me5) are important constituents of the neural circuitry responsible for jaw movements. Non N-methyl-D-aspartate (NMDA) glutamate receptors are a critical component of the brainstem circuitry responsible for reflex and centrally activated jaw movements; however, little is known about the expression of these receptors in neonatal oral motor circuitry. Receptor immunohistochemistry using affinity-purified polyclonal antibodies directed against GluR1, GluR2/3/4c, and GluR4, respectively, and a monoclonal antibody directed against the GluR2 subunit, were used in rats at postnatal day (P)1, P3, P5, P10, P19-21, P32-35, and P60 to describe the expression of the alpha-amino-d-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor in Mo5 and Me5 neurons. In Mo5, immunoreactivity was noted for all antibodies throughout the time frame sampled. Neurons in caudal portions of Me5 displayed immunoreactivity to each antibody except at P60 when GluR2 immunoreactivity was absent. Neurons located in rostral Me5 displayed GluR2/3/4c and GluR4 immunoreactivity throughout the time frame, GluR1 immunoreactivity emerged at P3 and a transient expression of GluR2 expression was observed between P10 and P32-35. The lack of labeling of some neurons in both regions, coupled with differences in temporal expression, suggests that there are differences in the AMPA receptor phenotype within and between Mo5 and Me5 during postnatal development. Differences in AMPA subunit composition suggest a complex role for AMPA-mediated glutamatergic neurotransmission in brainstem circuits controlling jaw movements. PMID- 11042595 TI - Extranuclear projections of rNST neurons expressing gustatory-elicited Fos. AB - Previous studies have demonstrated that gustatory stimulation evokes expression of the immediate-early gene, c-fos in the rostral division of the nucleus of the solitary tract (rNST) (Harrer and Travers [1996] Brain Res. 711:125-137; DiNardo and Travers [1997] J. Neurosci. 17:3826-3839; King et al. [1999] J. Neurosci. 19:3107-3121). The present investigation further defined the phenotype of those neurons by determining their projections, by using immunohistochemistry for the Fos protein and retrograde tracing with Fluoro-Gold. Tracer injections were made into the two major extranuclear targets of rNST, the parabrachial nucleus (PBN) and medullary reticular formation (RF). These structures are thought to play differential roles in higher-order discriminative and homeostatic (PBN) versus reflexive function (RF). After PBN injections, approximately 18% of the Fos-like immunoreactive (FLI) neurons were double-labeled; after RF injections the proportion was 9%. Because only a minority of FLI neurons appear to project to targets outside NST, this suggests that most of these cells have local, intranuclear projections. Comparable proportions of cells were double-labeled after sucrose or quinine, consistent with roles for both tastants in higher-order and reflexive function. On the other hand, regardless of stimulus, twice as many FLI neurons projected to the PBN as to the RF. This could suggest that more FLI neurons contribute to functions mediated by the ascending pathway. However, the results of a recent study prompted a different hypothesis: Because glossopharyngeal nerve section similarly devastates quinine-induced FLI and oral rejection but leaves discriminative function unimpaired, it was proposed that FLI neurons are more important in driving oral motor behavior than discrimination (King et al. [1999] J. Neurosci. 19:3107-3121). A plausible hypothesis for reconciling this apparent discrepancy is that many FLI neurons make local projections in rNST, that in turn give rise to RF connections. PMID- 11042597 TI - Erratum: yoon M-S, puelles L, redies C. 2000. Formation Of cadherin-expressing brain nuclei in diencephalic alar plate divisions. J. Comp. Neurol. 421:461-480 PMID- 11042596 TI - Clorgyline treatment elevates cortical serotonin and temporarily disrupts the vibrissae-related pattern in rat somatosensory cortex. AB - Manipulation of cortical serotonin (5-HT) levels in perinatal rodents produces significant alterations in the development of the layer IV cortical representation of the mystacial vibrissae. Monoamine oxidase A (MAO(A)) knockout mice have highly elevated cortical 5-HT and completely lack barrels in somatosensory cortex (S-I). The present study was undertaken to determine whether the effects on thalamocortical development seen in MAO(A) knockout mice can be replicated in perinatal rats treated with an MAO(A) inhibitor and, second, to determine whether these effects persist with continued treatment or after discontinuation of the drug. Littermates were injected with either clorgyline (5 mg/kg) or sterile saline five times daily. Clorgyline administration from birth to postnatal day (P) 6, 8, or 10 produced increases of 1,589.4 +/- 53.3%, 1660.2 +/- 43.1% and 1,700.5 +/- 84.5 %, respectively, in cortical 5-HT as compared with controls. Serotonin immunocytochemistry, 1,1;-dioctadecyl-3,3,3", 3; tetramethylindocarbocyanine perchlorate (DiI) labeling of thalamocortical afferents and Nissl and cytochrome oxidase staining of layer IV cellular aggregates demonstrated that clorgyline treatment from P0 to P6 produced a complete absence of any segmentation of vibrissae-related patches in S-I. However, continued treatment until P8 or P10 did not prevent the appearance of these patches. Animals treated with clorgyline from birth to P6 and killed on P8 or P10 had increases of 546.8 +/- 33.2% and 268.8 +/- 6.3% in cortical 5-HT and they had qualitatively normal vibrissae-related patterns in S-I. These results indicate that clorgyline treatment produces a transient disruption of vibrissae related patterns, despite the continued presence of elevated cortical 5-HT. PMID- 11042598 TI - Hydrophobicity parameter of diazines IV: a new hydrogen-accepting parameter of monosubstituted (di)azines for the relationship of partition coefficients in different solvent systems. AB - We recently proposed a new hydrogen-accepting scale, S(HA), for each member of the substituted (di)azine series on the basis of the heat of formation calculated under various dielectric environments by the COSMO method. In this paper, the S(HA) scale was used to examine relationships between log P(CL) (P(CL): CHCl(3)/H(2)O partition coefficient) and log P(oct) (P(oct): 1-octanol/H(2)O partition coefficient) for each of the 2-substituted pyridine (I), monosubstituted pyrazine (II), and pyrimidine (III) series. This S(HA) parameter worked nicely, representing the hydrogen-accepting effect of the solute molecule. A correlation equation with excellent quality, such as log P(CL) = a log P(oct) + sS(HA) + constant, was obtained for each series. We further defined the parameter S(HA/PY), derived from S(HA) values for the heterocyclic series by shifting the reference points to unsubstituted pyridine, to unify separately derived correlation equations. Thus, the correlation between log P(CL) and log P(oct) for all combined data of three series was derived by using a single equation as log P(CL) = a log P(oct) + sS(HA/PY) + constant. The S(HA) parameters were reasonably considered as being free-energy related, and the rationale for the hydrogen-bond acceptor scale was presented. PMID- 11042599 TI - Oxidation of HMG-CoA reductase inhibitors by tert-butoxyl and 1, 1-diphenyl-2 picrylhydrazyl radicals: model reactions for predicting oxidatively sensitive compounds during preformulation. AB - Hydrogen atom abstraction rate constants for the reaction of tert-butoxyl and 1,1 diphenyl-2-picrylhydrazyl (DPPH) radical with the HMG-CoA reductase inhibitors lovastatin, simvastatin, and statins I-IV were measured. This series of diene containing drugs is known to be prone to oxidation. The tert-butoxyl radical was generated by the thermolysis of di-tert-butylperoxyoxalate at 40 degrees C. A competitive kinetic method was used to determine the relative rate of hydrogen atom abstraction by tert-butoxyl radical to beta-scission. The absolute rate constants were calculated using the experimentally determined product ratios of t butanol to acetone and the known rate of beta-scission of tert-butoxyl radical. The rate constants for the reaction with DPPH radical were measured spectrophotometrically by monitoring the loss of DPPH radical as a function of substrate concentration. The rate constants correlate well with the structure of the molecules studied. These kinetic techniques allow for oxidatively sensitive compounds to be identified early in the drug development cycle. The tert-butoxyl radical, a strong hydrogen atom abstractor, is representative of the hydroxyl (. OH) and alkoxyl (. OR) radicals; in contrast the DPPH radical, a much weaker radical, is a good kinetic model for hydroperoxyl (. OOH) and peroxyl (. OOR) radicals. These kinetic methods can be used to quantitatively assess the lability of drug candidates towards reaction with oxygen-centered radicals at an early stage of development and facilitate the design of inhibiting strategies. PMID- 11042600 TI - Synthesis and solution properties of deferoxamine amides. AB - The poor membrane permeability and oral bioavailability of the iron chelating agent deferoxamine (DFO) mesylate result from the low octanol/water partition coefficient and high aqueous solubility. With the ultimate aim to improve biomembrane permeability while retaining the iron-binding ability of DFO, a series of more lipophilic amides were prepared by reacting the terminal primary amino group with fatty and aromatic acid chlorides or anhydrides. Octanol/water partition coefficients and equilibrium solubilities of these analogs in solvents, chosen to delineate physicochemical interactions, were determined as a function of temperature. Solid-state properties were evaluated by calorimetry. All DFO amide derivatives had higher melting points, indicating that derivatives formed strong intermolecular interactions in the solid phase. Formamidation of the primary amine of deferoxamine resulted in a 200-fold increase in the octanol/water partition coefficient and reduced aqueous solubility at least 2000 fold compared with the parent molecule. The partition coefficient increased and aqueous solubility decreased 2-fold with the addition of each methylene group in the homologous series of aliphatic amides. Solubilities of the derivatives in water-saturated octanol and hexane showed irregular profiles as a function of increasing aliphatic chain length that were attributed to intermolecular packing in the solid state. The temperature dependence of the partition coefficients was interpreted to indicate that interfacial transfer of the deferoxamine amides was, in part, affected by an apparent diminished ability to form energetically favorable interactions in the water-saturated organic phase. PMID- 11042601 TI - Scanning electrochemical microscopy of iontophoretic transport in hairless mouse skin. Analysis of the relative contributions of diffusion, migration, and electroosmosis to transport in hair follicles. AB - Scanning electrochemical microscopy (SECM) is used to measure spatially localized diffusive and iontophoretic transport rates in hairless mouse skin. Molecular fluxes within individual hair follicles are quantified by measuring the rate at which redox-active probe molecules emerge from the follicle. The influence of an applied current on the flux of an anion (ascorbate), a cation (ferrocenylmethyltrimethylammonium), and a neutral molecule (acetaminophen) is used to determine the contributions of diffusion, migration, and electroosmosis to iontophoretic transport. The direction of electroosmotic transport is consistent with hair follicles possessing a net negative charge at neutral pH. Electroosmosis results in a modest increase in the transport rate of the neutral molecule (a factor of approximately 2.4x at an iontophoretic current density of 0.1 mA/cm(2)). Larger enhancements in the flux of the electrically charged species are associated with migration. The electroosmotic flow velocity within hair follicles is established to be 0.5 (+/-0.1) microm/s at 0.1 mA/cm(2), independent of the electrical charge of permeant. The net volume flow rate across skin resulting from electroosmosis in hair follicles is estimated to be 0.3 microL/cm(2)h. The results suggest that hair follicles are a significant pathway for electroosmotic solution flow during iontophoresis. The radius of the hair follicle openings in hairless mouse skin is measured to be 21 +/- 5 microm. PMID- 11042602 TI - Enhancing the immunogenicity of liposomal hepatitis B surface antigen (HBsAg) by controlling its delivery from polymeric microspheres. AB - Microencapsulated liposome systems (MELs) were investigated as a potential immunization carrier for a recombinant 22-nm hepatitis B surface antigen (HBsAg) particle. MELs were prepared by first entrapping the HBsAg particles within liposomes composed of phosphatidylcholine:cholesterol (1:1 molar ratio), which were then encapsulated within alginate-poly(L-lysine) (PLL) hydrogel microspheres. The entrapped HBsAg particles retained immunoreactivity, as judged by an enzyme-linked immunosorbent assay (ELISA). Direct imaging of HBsAg particles and HBsAg incorporated into liposomes by cryo-transmission electron microscopy (cryo-TEM) indicated that HBsAg is embedded in the liposomal membrane. The antigenic particles were released from MELs mainly within the context of liposomes. The release rates in vitro and in vivo depended on the molecular weight of PLL used for MEL coating; MELs-214, coated with 214 kDa PLL, released the liposomal HBsAg at much higher rates than MELs-25, which was coated with 25 kDa PLL. Concomitantly, the specific anti-HBsAg titers in mice receiving HBsAg in MELs-214 were higher than those induced by MELs-25. MELs-214 were more efficient than conventional liposomes or alum in eliciting higher and prolonged antibody levels in mice. The ability of MELs to provide an HBsAg depot as well as a sustained release of liposomal HBsAg suggests that these carriers may be an ideal immunoadjuvant. PMID- 11042603 TI - Physicomechanical properties of biodegradable poly(D,L-lactide) and poly(D,L lactide-co-glycolide) films in the dry and wet states. AB - The objective of this study was to investigate the mechanical properties (% elongation and puncture strength) of poly(D,L-lactide) (PLA) and poly(D,L-lactide co-glycolide) (PLGA) films as a function of exposure time to an aqueous medium and to correlate the mechanical properties to the degradation/erosion of the polymer as a function of the type of polymer [PLA, weight-average molecular weight (M(W)) 270,300, or PLGA 50:50, M(W) 56,500], the type of plasticizer [(triethyl citrate (TEC) or acetyltributyl citrate (ATBC)], and the exposure time to pH 7.4 phosphate buffer. The glass transition temperature of the films was measured by differential scanning calorimetry (DSC), the molecular weight by size exclusion chromatography (SEC), and the polymer erosion and hydration gravimetrically. The mechanical properties were strongly affected by the type of polymer and plasticizer. PLGA films showed a faster loss of mechanical integrity. TEC, the water-soluble plasticizer, leached from the films, resulting in major differences in the mechanical properties (flexibility) when compared with films plasticized with the more permanent, water-insoluble ATBC. A significant difference in M(W) decrease was seen between plasticizer-free and plasticizer containing PLA films, but not for PLGA films. Plasticized PLA films, which were above their glass transition temperature in the rubbery state, showed a faster decrease in M(W) than plasticizer-free PLA ones, which were in the glassy state. The plasticizer addition to the lower M(W) PLGA did not enhance the polymer degradation; the plasticizer-free PLGA was already in the rubbery state. Major differences between the two polymers were also seen in the mass loss and the water uptake studies. After 4 weeks, the mass loss was between 2.6 and 7.0% and the water uptake between 10.1 and 21.1% for PLA films, whereas for PLGA films, the mass loss was between 40.3 and 51.3% and the water uptake between 221.9 and 350.6%. 2000 Wiley-Liss, Inc. PMID- 11042604 TI - Evaluating hydrogen-bond donor strength. AB - A parameter that measures hydrogen-bond donor (HBD) strength of solutes is useful in modeling many biological interactions. The solvents octanol and chloroform, have about equal HBD strength and thus will accommodate the hydrogen-bond acceptor (HBA) groups in solutes about equally well. Because the solvent octanol has a strong acceptor oxygen, solutes with HBD groups will favor it over chloroform on that basis. With its eight alkane carbons, octanol also favors solutes with a significant amount of alkane character, a property referred to in this paper as 'excess alkane affinity' (XAA). On the other hand, it is easier to form a cavity in the solvent chloroform, so larger solutes tend to favor that solvent. After allowing for XAA and molecular volume, the difference between log P(oct) and log P(clf) is a measure of the effective sum of HBD. This value is given the symbol epsilonalpha and appears to be on the same scale as Abraham's summation operatoralpha(2)(H). PMID- 11042605 TI - Commentary: using the convection-dispersion model and transit time density functions in the analysis of organ distribution kinetics. AB - The convection-dispersion model and its extended form have been used to describe solute disposition in organs and to predict hepatic availabilities. A range of empirical transit-time density functions has also been used for a similar purpose. The use of the dispersion model with mixed boundary conditions and transit-time density functions has been queried recently by Hisaka and Sugiyama in this journal. We suggest that, consistent with soil science and chemical engineering literature, the mixed boundary conditions are appropriate providing concentrations are defined in terms of flux to ensure continuity at the boundaries and mass balance. It is suggested that the use of the inverse Gaussian or other functions as empirical transit-time densities is independent of any boundary condition consideration. The mixed boundary condition solutions of the convection-dispersion model are the easiest to use when linear kinetics applies. In contrast, the closed conditions are easier to apply in a numerical analysis of nonlinear disposition of solutes in organs. We therefore argue that the use of hepatic elimination models should be based on pragmatic considerations, giving emphasis to using the simplest or easiest solution that will give a sufficiently accurate prediction of hepatic pharmacokinetics for a particular application. PMID- 11042606 TI - Problems of mixed boundary conditions for convection-dispersion models in the analysis of local pharmacokinetics. PMID- 11042607 TI - The effect of serum albumin on the aggregation state and toxicity of amphotericin B. AB - Studies have shown that the dose-limiting toxicity of amphotericin B (AmB), a key drug for systemic mycoses, depends on its self-aggregation state. In a step toward understanding the various factors in blood mediating the toxicity of AmB, we have investigated the effect of serum albumin, the most abundant plasma protein, on the aggregation state of AmB using absorption spectroscopy. The critical aggregation concentration (CAC) of AmB, which coincides with its concentration at the onset of toxicity (hemolysis), was 1.1 microM, but rose in proportion to the level of serum albumin (1.0 to 4.0% w/v). The CAC of AmB was 8.0 microM at 4.0% w/v serum albumin, which is considerably higher than peak therapeutic levels of AmB in plasma (i.e., 2.0 microM). Serum albumin (4.0% w/v) lowered the degree of aggregation of AmB (size of aggregates) above the CAC and increased its solubility. The results suggest that serum albumin attenuates the toxicity of AmB at a membrane level by affecting its aggregation state. In this way, serum albumin in blood may balance deleterious effects of AmB mediated by serum low-density lipoproteins. PMID- 11042608 TI - Inhibitory effect of statins on fetal bovine serum-induced proliferation of rat cultured mesangial cells and correlation between their inhibitory effect and transport characteristics. AB - Mesangial cells play an important role in physiologic functions, including the regulation of glomerular filtration, and as a pathogenic factor for proliferative glomerulonephritis. We compared the potencies of the inhibitory effects of simvastatin acid, lovastatin acid, and pravastatin on fetal bovine serum (FBS) induced proliferation of rat cultured mesangial cells, and examined the correlation between their inhibitory effects and intracellular concentrations. We also investigated the transport of the statins in the cells, and whether or not their intracellular concentrations were determined by their transport characteristics. It appeared that the growth inhibitory effects on FBS-induced proliferation of mesangial cells of simvastatin acid and lovastatin acid were approximately the same, but that of pravastatin was extremely weak compared with the others. The growth inhibitory effects of these agents were suggested to depend, at least in part, on the amount incorporated intracellularly. Simvastatin acid, lovastatin acid, and pravastatin appeared to be taken up by mesangial cells via a common carrier, the uptake capacity being determined by their lipophilicity. Therefore, it was thought that the growth inhibitory effects of the statins partially depended on their carrier-mediated uptake by mesangial cells. PMID- 11042609 TI - Conformational stability of human interferon-gamma on association with and dissociation from liposomes. AB - The integrity of a therapeutic protein has to be safeguarded when formulated in delivery systems such as liposomes. In this study, we investigated the conformational stability of recombinant human interferon gamma (hIFNgamma) on association with and after dissociation from liposomal bilayers using circular dichroism (CD) and steady-state fluorescence spectroscopy as well as time resolved fluorescence methodology. We used hIFNgamma adsorption to and desorption from empty liposomes as a model for hIFNgamma-containing liposomes prepared via the film hydration method. CD studies indicated that no changes in the secondary and tertiary protein structure occur during and after interaction of hIFNgamma with the liposomes. Steady-state fluorescence emission spectra of untreated and liposome-desorbed hIFNgamma revealed that the environment of the sole Trp residue was not affected by the adsorption/desorption process. The Trp-36 residue remained fully quenchable by acrylamide after desorption of hIFNgamma from the liposomes. Time-resolved fluorescence studies were conducted to probe the local environment and the mobility of Trp-36 before, during, and after interaction of hIFNgamma with the liposomal membrane. Differences in rotational correlation time between free and liposomal hIFNgamma were attributed to immobilization of the protein on adsorption to the liposome bilayer. Disparities were detected between the average lifetimes of liposome-adsorbed hIFNgamma and hIFNgamma-liposomes, indicating that subtle changes in the Trp-36 environment took place during preparation of the liposomes via the film hydration method compared with the adsorption of hIFNgamma to the liposome surface. The results of this study indicate that association of hIFNgamma with negatively charged liposomes results in minimal changes in the secondary and tertiary structure of the protein. We conclude that all techniques used point to a full retention or restoration of the protein conformation after desorption from the liposomes. PMID- 11042610 TI - Estimation of the effect of NaCl on the solubility of organic compounds in aqueous solutions. AB - The Setschenow constant, K(salt), of a nonelectrolyte in a NaCl solution is shown to be related to the logarithm of its octanol-water partition coefficient, log K(ow), determined by K(salt) = A log K(ow) + B, where K(ow) is the octanol-water partition coefficient of the solute and the coefficients A and B are constants. The values of A and B were empirically determined from literature data for 62 organic compounds and validated for a test set of 15 compounds including several drugs. PMID- 11042611 TI - Algorithms for quantitation of protein expression variation in normal versus tumor tissue as a prognostic factor in cancer: Met oncogene expression, and breast cancer as a model. AB - BACKGROUND: Immunohistochemistry and immunofluorescence (IF) assays frequently rely on subjective observer evaluation for grading. The aim of our study was to develop an objective quantitative index based on confocal laser scanning microscopy (CLSM) and image analysis of an IF assay to determine alteration in protein expression levels in normal versus tumor tissue. The relative levels of Met expression, a prognostic factor in breast cancer, were used as a model for evaluating image analysis algorithms. METHODS: Primary human breast cancer biopsies were collected. Sections containing tumor and adjacent uninvolved normal regions were immunostained for Met and digital images were acquired by CLSM. Subsequently, the digital data were manipulated using several different algorithms to calculate prognostic indexes. The results were correlated with the clinical outcome to determine the prognostic value of these indexes. RESULTS: Different algorithms were used to obtain quantitative indexes to evaluate the relative levels of Met expression. We report a statistical correlation between patient prognosis and relative Met level in normal versus tumor tissue as determined by three distinct algorithms using Kaplan-Meier analysis (log-rank): calculations based on intensity levels differences DV (P = 0.002), DIntensity (P = 0.014), and entropy divergence (Dentropy; P = 0.0023). CONCLUSIONS: Using adjacent normal tissue as an internal reference, a quantitative index of tumor Met level divergence can be objectively determined to have a prognostic value. Moreover, this methodology can be used for other proteins in a variety of different diseases. PMID- 11042612 TI - Measurement of markers for breast cancer in a model system using laser scanning cytometry. AB - BACKGROUND: A variety of markers, including Ki67, estrogen receptors (ER), and progesterone receptors (PgR), are frequently measured in fine needle aspirates (FNA) from human breast carcinomas. We used a human breast carcinoma cell line, MCF7, as a model system to investigate the use of laser scanning cytometry (LSC) for the measurement of these markers. Additionally, we measured the number of apoptotic cells. METHODS: Cells were treated with drugs to vary the expression of markers and the number of apoptotic cells. They were then fixed on microscope slides. For LSC, the cells were stained for the different markers with fluorescein using immunofluorescence and for apoptotic cells using the TUNEL assay. The nuclei were counterstained with propidium iodide. A parallel set of slides was stained using horseradish peroxidase and diaminobenzidine and scored manually by conventional light microscopy. RESULTS: The results from the LSC closely paralleled those obtained by manual scoring of immunohistochemical stains. CONCLUSIONS: It should be possible to use LSC for the routine measurement of nuclear markers in FNAs from human breast carcinomas. PMID- 11042613 TI - Cell heterogeneity and subpopulations in solid tumors characterized by simultaneous immunophenotyping and DNA content analysis. AB - BACKGROUND: Heterogeneity in human malignant tumors is a well-described phenomenon and of interest with regard to subpopulations with differences in clonality, metastatic potential, and response to therapy under different treatment regimes. The aim of this study was the simultaneous characterization of surface markers and DNA content of solid tumors to identify tumor cell subpopulations and to study the association between the expression of antigens and DNA content. METHODS: In the present study, six different malignant tumors grown as xenografts in nude mice were characterized by five-parameter flow cytometry. Immunophenotyping was performed using a variety of direct fluorescence conjugated antibodies. In all cases, simultaneous detection of DNA content was done after staining with 7-aminoactinomycin D. RESULTS: Tumor cells were characterized by light scatter properties, antigen expression, and DNA content. Tumor cell heterogeneity, subpopulations, and DNA content-dependent antigen expression were identified. CONCLUSIONS: This method offers the possibility of characterizing solid tumors according to their immunophenotype and DNA content. The results obtained can be used to identify changes in immunophenotypic and DNA profiles of tumor cell populations before and after therapy and might be useful to define parameters predictive for response to therapy. PMID- 11042614 TI - Fluorescence lifetime imaging of nuclear DNA: effect of fluorescence resonance energy transfer. AB - BACKGROUND: DNA fluorescence dyes have been used to study DNA dynamics, chromatin structure, and cell cycle analysis. However, most microscopic fluorescence studies of DNA use only steady-state measurements and do not take advantage of the additional information content of the time-resolved fluorescence. In this paper, we combine fluorescence imaging of DNA with time-resolved measurements to examine the proximity of donors and acceptors bound to chromatin. METHODS: We used frequency-domain fluorescence lifetime imaging microscopy to study the spatial distribution of DNA-bound donors and acceptors in fixed 3T3 nuclei. Over 50 cell nuclei were imaged in the presence of an AT-specific donor, Hoechst 33258 (Ho), and a GC-specific acceptor, 7-aminoactinomycin D (7-AAD). RESULTS: The intensity images of Ho alone showed a spatially irregular distribution due to the various concentrations of DNA or AT-rich DNA throughout the nuclei. The lifetime imaging of the Ho-stained nuclei was typically flat. Addition of 7-AAD decreased the fluorescence intensity and lifetime of the Ho-stained DNA. The spatially dependent phase and modulation values of Ho in the presence of 7-AAD showed that the Ho decay becomes nonexponential, as is expected for a resonance energy transfer (RET) with multiple acceptors located over a range of distances. In approximately 40 nuclei, the intensity and lifetime decrease was spatially homogeneous. In approximately 10 nuclei, addition of 7-AAD resulted in a spatially nonhomogeneous decrease in intensity and lifetime. The RET efficiency was higher in G(2)/M than in G(0/1) phase cells. CONCLUSIONS: Because RET efficiency depends on the average distance between Ho and 7-AAD, data suggest that the heterogeneity of lifetimes and spatial variation of the RET efficiency are caused by the presence of highly condensed regions of DNA in nuclei. PMID- 11042615 TI - Membrane permeability changes induce hyperpolarization in transformed lymphoid cells under high-density culture conditions. AB - BACKGROUND: Membrane potential changes in cells from the human lymphoid B cell line, JY, evoked by increasing cell density in culture were investigated, as data published on other cell types are controversial. An attempt was also made to clear the underlying mechanism. METHODS: Nonadherent JY cells were isolated from high-density plateau-phase cultures (type A cells), medium-density log-phase cultures (type B cells), and low-density lag-phase cultures (type C cells). They were analyzed for transmembrane potential, intracellular free concentration of potassium and sodium, membrane permeability for monovalent cations, cell cycle distribution by measuring DNA content, and glucose uptake. RESULTS: C type cells proved to be relatively depolarized (-41 +/- 3 mV) and cells obtained from the highest density cultures hyperpolarized (-60 +/- 3 mV). Intracellular concentrations ([K](i) = 92-97 mM and [Na](i) = 34-35 mM) were almost identical for each type of cell. The sodium/potassium permeability constant ratio in the A and C type of cells was 0.047 and 0.094, respectively. High-density culture conditions resulted in a pronounced G(1)-phase arrest. CONCLUSIONS: Differences in the membrane potential values induced by high-density culture conditions were maintained by changes in the membrane permeability for the monovalent cations. PMID- 11042616 TI - One-round determination of seven leukocyte subsets in rhesus macaque blood by flow cytometry. AB - BACKGROUND: Rhesus macaques are frequently used in biomedical research as experimental models for studying infectious diseases and for preclinical vaccination trials. The infection of these monkeys with simian immunodeficiency viruses (SIV) or simian-human immunodeficiency viruses (SHIV) reproduces the clinical and immunological characteristics of human infection by human immunodeficiency virus (HIV). Evolution of the immune response in the infected animals is generally analyzed by determining the lymphocyte subsets on blood samples using flow cytometry but requiring multiple, blood consuming, determinations. METHODS: Cell subsets present in whole-blood samples were labeled with a combination of anti-human monoclonal antibodies to CD2, CD20, CD4, CD8, and CD14 coupled to FITC or PE and analyzed by flow cytometry. RESULTS: In one round, we obtained the precise determination of macaque blood cell composition by flow cytometry. Monocytes, granulocytes, eosinophils, B lymphocytes, helper, and cytotoxic T lymphocytes were distinguished. Results obtained correlated strongly with those obtained with conventional blood cell differential systems and with separate staining of lymphocytes. The analysis of blood from healthy rhesus macaques and SHIV-infected animals demonstrated the accuracy of the determination even in very pathological situations such as macaques with simian AIDS. CONCLUSIONS: Our method allows fast determination of the blood cell composition and will be particularly useful to evaluate the cell subset evolution of macaques involved in large-scale experimental trials. PMID- 11042617 TI - Rapid DNA fingerprinting of pathogens by flow cytometry. AB - BACKGROUND: A new method for rapid discrimination among bacterial strains based on DNA fragment sizing by flow cytometry is presented. This revolutionary approach combines the reproducibility and reliability of restriction fragment length polymorphism (RFLP) analysis with the speed and sensitivity of flow cytometry. METHODS: Bacterial genomic DNA was isolated and digested with a rare cutting restriction endonuclease. The resulting fragments were stained stoichiometrically with PicoGreen dye and introduced into an ultrasensitive flow cytometer. A histogram of burst sizes from the restriction fragments (linearly related to fragment length in base pairs) resulted in a DNA fingerprint that was used to distinguish among different bacterial strains. RESULTS: Five different strains of gram-negative Escherichia coli and six different strains of gram positive Staphylococcus aureus were distinguished by analyzing their restriction fragments with DNA fragment sizing by flow cytometry. Fragment distribution analyses of extracted DNA were approximately 100 times faster and approximately 200,000 times more sensitive than pulsed-field gel electrophoresis (PFGE). When sample preparation time is included, the total DNA fragment analysis time was approximately 8 h by flow cytometry and approximately 24 h by PFGE. CONCLUSIONS: DNA fragment sizing by flow cytometry is a fast and reliable technique that can be applied to the discrimination among species and strains of human pathogens. Unlike some polymerase chain reaction (PCR)-based methods, sequence information about the bacterial strains is not required, allowing the detection of unknown, newly emerged, or unanticipated strains. PMID- 11042618 TI - Flow cytometry as a strategy to study the endosymbiosis of algae in Paramecium bursaria. AB - BACKGROUND: The stable symbiotic association between Paramecium bursaria and algae is of interest to study such mechanisms in biology as recognition, specificity, infection, and regulation. The combination of algae-free strains of P. bursaria, which have been recently established by treating their stocks of green paramecia with herbicide paraquat (Hosoya et al.: Zool Sci 12: 807-810, 1995), with the cloned symbiotic algae isolated from P. bursaria (Nishihara et al.: Protoplasma 203: 91-99, 1998), provides an excellent clue to gain fundamental understanding of these phenomena. METHODS: Flow cytometry and light microscopy have been employed to characterize the algal cells after they have been released from the paramecia by ultrasonic treatment. Algal optical properties such as light scattering and endogenous chlorophyll fluorescence intensity have been monitored for symbiotic and free-living strains, and strains at stages of interaction with a host. RESULTS: Neither algal morphology nor chlorophyll content has been found to be altered by sonication of green paramecia. This fact allows to interpret in adequate degree changes in the optical properties of symbiont that just has been released from the association with a host (decreased forward light scatter and chlorophyll fluorescence signals). Optical characterization of both symbiotic and free-living algal strains with respect to their ability to establish symbioses with P. bursaria showed that chlorophyll content per cell volume seems to be a valuable factor for predicting a favorable symbiotic relationship between P. bursaria and algae. CONCLUSIONS: Flow cytometry combined with algae-free paramecia and cloned symbiotic algae identifies algal populations that may be recognized by host cells for the establishment of symbioses. PMID- 11042619 TI - A novel two-color flow cytometric assay for the detection of Cryptosporidium in environmental water samples. AB - BACKGROUND: Cryptosporidium is an important waterborne pathogen. Detection of Cryptosporidium in concentrated water samples depends on oocyst isolation using immunomagnetic separation (IMS) and/or fluorescence-activated cell sorting (FACS), followed by confirmation using immunofluorescence staining (IFA) and fluorescence microscopy. These methods require highly trained microscopists for oocyst identification and confirmation. Analysis is hampered due to the presence of autofluorescent particles coupled with particles binding nonspecifically with the monoclonal antibodies (mAbs) used for detection. Flow cytometry (FCM) has the potential to be a more specific method for oocyst detection, but such a system would require more than one selection parameter. METHODS: Various mAbs from commercial suppliers were paired with CRY104-PE and evaluated. The mAb combination that best discriminated stained oocyst from detritus was optimized and compared to Cryptosporidium detection utilizing one-color IFA/FACS. RESULTS: A highly specific two-color assay employing the IgG(1) mAb CRY104 was developed. The assay resulted in reductions, up to 20-fold, in the number of non Cryptosporidium particles detected. The addition of a second selection parameter improved microscopic analysis times and simplified oocyst confirmation by microscopists. CONCLUSIONS: A two-color assay employing competing surface mAbs reduces the number of fluorescent particles sorted, thus improving FCM detection methods for Cryptosporidium. PMID- 11042620 TI - Flow cytometry to evaluate Theileria sergenti parasitemia using the fluorescent nucleic acid stain, SYTO16. AB - BACKGROUND: Although the infection of Theileria sergenti is demonstrated by intraerythrocytic localization of this parasite, much time and labor are necessary in order to determine this. We applied flow cytometry to evaluate T. sergenti parasitemia using the fluorescent nucleic acid stain method. METHODS: Peripheral blood samples from cattle infected with T. sergenti were stained with a membrane-permeable fluorescent nucleic acid stain, SYTO16, and hydroethidine (HE). Stained parasitized erythrocytes were measured by a flow cytometer equipped with a single argon laser operating at 488 nm. RESULTS: SYTO16-stained intraerythrocytic parasites were detected on the FL1 (525 nm) and parasitized cells were separated completely from unparasitized cells. However, HE-stained erythrocytes could not be divided clearly into parasitized and unparasitized cells. SYTO16-stained parasites were reproducibly detected at a percentage above 0.1%. Contaminating leukocytes, which were indicated by CD18-positive cells, were eliminated from the analysis by narrowing the light scatter gate of the erythrocyte fraction. A correlation (r = 0.983) between the percentage of SYTO16 positive cells and parasitemia in grazing cattle was observed. CONCLUSIONS: Flow cytometric detection using SYTO16 is a rapid and reliable method of monitoring parasitemia in T. sergenti-infected cattle. PMID- 11042621 TI - Phycobiliprotein-Fab conjugates as probes for single particle fluorescence imaging. AB - BACKGROUND AND METHODS: Single particle fluorescence imaging (SPFI) is a recently developed method that has provided a powerful approach to observing receptor movement and associations at high spatial resolution. It provides a noninvasive alternative to the existing biochemical techniques. It can also quantify and resolve molecular interactions at the cell surface at a nanometer scale. Probes that have been used in the past to study mobility and associations of cell surface receptors have many limitations. These include concerns about the specificity of the probes, the possibility that their size interferes with the receptor once bound to it, the nonuniform fluorescence, and the questionable stoichiometry. RESULTS: In this study, we have generated phycobiliprotein-Fab conjugates, and have shown that they are a significant advance on existing probes for SPFI studies. They are small in size, highly specific, highly fluorescent, of known stoichiometry, photostable, emit uniform fluorescence, and are generally well defined. CONCLUSIONS: It is highly important that when studying receptor mobility or associations, fully characterized probes are used. Phycoerythrin(PE) Fab probes provide us with the perfect tool for SPFI, and a system with a wide range of applicability to study any cell surface receptor against which a monoclonal antibody exists. PMID- 11042622 TI - Formal approaches to safety monitoring of clinical trials in life-threatening conditions. AB - Large clinical trials in life-threatening conditions are usually conducted under the surveillance of a Data and Safety Monitoring Board (DSMB), whose remit is to protect the ethical and safety interests of the patients. The purpose of this paper is to describe a formal approach to safety monitoring, using a sequential safety procedure to aid the decisions made by the DSMB. This procedure is designed to recommend termination of the study as soon as evidence that the experimental treatment is worse than the control in terms of the primary safety response is so strong that it is unethical to proceed. The use of this formal sequential procedure enables probabilities of the study stopping erroneously and stopping correctly, under various degrees of experimental treatment disadvantage, to be examined. Also scenarios depicting data sets which lead to continuing or stopping can be presented. Such explorations are useful in encouraging all DSMB members to consider carefully, prior to the start of a study, the conditions under which they would seriously wish to consider termination. The implementation of these methods is described for three recently completed trials in which it has been used. Finally, our current recommendations for the design of these procedures, arising from these and other similar experiences, are given. PMID- 11042623 TI - Tests for homogeneity of the risk ratio in a series of 2x2 tables. AB - We often apply the risk ratio to measure the strength of a causal relationship between a suspected risk factor and a disease of interest. In this paper we consider testing the homogeneity of risk ratio over a series of 2x2 tables. In addition to the classical weighted least squares (CWLS) test procedure, we consider two test procedures using simple transformations of the CWLS statistic and develop three other asymptotically weighted test procedures. On the basis of Monte Carlo simulation, we conclude that the commonly-used CWLS test procedure is generally conservative, especially when the number of 2x2 tables is large and the mean group size per table is moderate or small. We further find that two of the test procedures discussed here can not only generally outperform the CWLS test procedure, but also perform well in a variety of situations considered in this paper. Finally, we illustrate the use of these testing procedures with an example of six randomized trials that assess the effects of aspirin on the prevention of death in post-myocardial infarction patients. PMID- 11042624 TI - Confidence intervals for the risk ratio under cluster sampling based on the beta binomial model. AB - In cohort studies, the risk ratio (RR) is one of the most commonly used epidemiologic indices to quantify the effect of a suspected risk factor on the probability of developing a disease. When we employ cluster sampling to collect data, an interval estimator that does not account for the intraclass correlation between subjects within clusters is likely inappropriate. In application of the beta-binomial model to account for the intraclass correlation, we develop four asymptotic interval estimators of the RR, which are direct extensions of some recently developed estimators for independent binomial sampling. We then use Monte Carlo simulation to evaluate the finite-sample performance of these four interval estimators in a variety of situations. We find that the estimator using the logarithmic transformation generally performs well and is preferable to the other three estimators in most of the situations considered here. Finally, we include an example from a study of an educational intervention with emphasis on behaviour change to illustrate the use of the estimators developed in this paper. PMID- 11042625 TI - Goodness-of-fit statistics for age-specific reference intervals. AB - The age-specific reference interval is a commonly used screening tool in medicine. It involves estimation of extreme quantile curves (such as the 5th and 95th centiles) of a reference distribution of clinically normal individuals. It is crucial that models used to estimate such intervals fit the data extremely well. However, few procedures to assess goodness-of-fit have been proposed in the literature, and even fewer have been evaluated systematically. Here we consider procedures based on the distribution of the Z-scores (standardized residuals) from a model and on Pearson chi(2) statistics for observed and expected counts in groups defined by age and the estimated reference centile curves. Two of the procedures (Q and grid tests) are mainly inferential, whereas the third (permutation bands and B-tests) is essentially graphical. We obtain approximations to the null distributions of several relevant test statistics and examine their size and power for a range of models based on real data sets. We recommend Q-tests in all situations where Z-scores are available since they are general, simple to calculate and usually have the highest power among the three classes of test considered. For the cases considered the grid tests are always inferior to the Q- and B- tests. PMID- 11042626 TI - Permutation tests for comparing marginal survival functions with clustered failure time data. AB - We propose a class of two-sample non-parametric permutation tests to compare the marginal survival distributions of two groups when the failure times are correlated within cluster, with clusters nested within each group. The permutation distribution effectively takes into account the correlation between failure times within a cluster. The method is able to handle data with clusters of either fixed or variable sizes. Moreover, this class of test statistics is sensitive to various alternatives. The size and power of the proposed tests are assessed by a series of simulation studies. The method is illustrated by application to data from the Hypertension Detection and Follow-up program trial. PMID- 11042627 TI - Maximum likelihood estimation for longitudinal data with truncated observations. AB - We obtain maximum likelihood estimates of the parameters when the observations on the response variable in a repeated measures design are truncated above a cutpoint. The maximum likelihood equations are solved iteratively using an EM like procedure. It is observed that these estimates have smaller mean squared error than recently proposed iterative weighted least-squares estimates. The results are applied to data arising from a study of dioxin elimination in Air Force veterans. Published in 2000 by John Wiley & Sons, Ltd. PMID- 11042628 TI - Use of an angular transformation for ratio estimation in cost-effectiveness analysis. AB - Economic evaluations of medical technologies involve a consideration of both costs and clinical benefits, and an increasing number of clinical studies include a specific objective of assessing cost-effectiveness. These studies measure the trade-off between costs and benefits using the cost-effectiveness ratio (CE ratio), which is defined as the net incremental cost per unit of benefit provided by the candidate therapy. In this paper we review the statistical methods which have been proposed for estimating 95 per cent confidence intervals for cost effectiveness ratios. We show that the use of an angular transformation of the standardized ratio stabilizes the variance of the estimated CE ratio, and provides a clearer interpretation of study results. An estimate of the 95 per cent confidence interval for the CE ratio in the transformed scale is easily made using the jack-knife or bootstrap. The available methods are compared using data from a long term study of mortality in patients with congestive heart failure. PMID- 11042629 TI - Estimation of current leukaemia-free survival following donor lymphocyte infusion therapy for patients with leukaemia who relapse after allografting: application of a multistate model. AB - Allogeneic bone marrow transplantation has become routine treatment for selected patients with leukaemia and other haematological disorders. A standard measure of the efficacy of this treatment for patients is assessment of leukaemia-free survival, that is, the probability of being alive and in remission at different time intervals after transplant. During the last 10 years a new approach to managing patients who relapse after allografting was developed - namely infusion of lymphocytes (white blood cells) collected from the original donor. Such donor lymphocyte infusions (DLI) are highly effective in restoring complete remission and it appears that these remissions are durable. Consequently estimates of long term success of allografting must consider salvage of initial relapses with DLI. To do so a new estimator of treatment efficacy, called 'current leukaemia-free survival' is sometimes used in the medical literature. This curve attempts to estimate the probability that a patient is alive in an original remission or in subsequent remission after treatment with DLI at a given time after transplant. Here we show that this estimator does not actually estimate the probability of interest and we use a multi-stage model to develop a new and more appropriate estimator for 'current leukaemia-free survival'. PMID- 11042630 TI - M. D. deB. Edwardes, 'The generalization of the odds ratio, risk ratio and risk difference to r x k tables'. Statistics in Medicine 2000; 19(14): 1901-1914. AB - The original article to which this Correction refers was published in Statistics in Medicine 2000 19(14): 1901-1914. PMID- 11042631 TI - Interventional MRI-challenge for radiology. PMID- 11042632 TI - Intraoperative magnetic resonance image guidance in neurosurgery. AB - Recently, there has been a burgeoning interest in the use of image-guided navigation to improve the safety and effectiveness of neurosurgical procedures. The intraoperative use of magnetic resonance imaging (MRI) provides the most accurate guidance available. This report discusses the hardware and software improvements that have made intraoperative MRI a reality and describes the use of this technology for neurosurgical intraoperative guidance. PMID- 11042633 TI - Magnetic resonance temperature imaging for guidance of thermotherapy. AB - Continuous thermometry during a hyperthermic procedure may help to correct for local differences in heat conduction and energy absorption, and thus allow optimization of the thermal therapy. Noninvasive, three-dimensional mapping of temperature changes is feasible with magnetic resonance (MR) and may be based on the relaxation time T(1), the diffusion coefficient (D), or proton resonance frequency (PRF) of tissue water. The use of temperature-sensitive contrast agents and proton spectroscopic imaging can provide absolute temperature measurements. The principles and performance of these methods are reviewed in this paper. The excellent linearity and near-independence with respect to tissue type, together with good temperature sensitivity, make PRF-based temperature MRI the preferred choice for many applications at mid to high field strength (>/= 1 T). The PRF methods employ radiofrequency spoiled gradient-echo imaging methods. A standard deviation of less than 1 degrees C, for a temporal resolution below 1 second and a spatial resolution of about 2 mm, is feasible for a single slice for immobile tissues. Corrections should be made for temperature-induced susceptibility effects in the PRF method. If spin-echo methods are preferred, for example when field homogeneity is poor due to small ferromagnetic parts in the needle, the D- and T(1)-based methods may give better results. The sensitivity of the D method is higher that that of the T(1) methods provided that motion artifacts are avoided and the trace of D is evaluated. Fat suppression is necessary for most tissues when T(1), D, or PRF methods are employed. The latter three methods require excellent registration to correct for displacements between scans. PMID- 11042634 TI - Interventional MRA and intravascular imaging. AB - Several attributes make magnetic resonance imaging (MRI) attractive for guidance of intravascular therapeutic procedures, including high soft tissue contrast, imaging in arbitrary oblique planes, lack of ionizing radiation, and the ability to provide functional information, such as flow velocity or flow volume per unit time, in conjunction with morphologic information. For MR guidance of vascular interventions to be safe, the interventionalist must be able to visualize catheters and guidewires relative to the vascular system and surrounding tissues. A number of approaches for rendering instruments visible in an MR environment have been developed, including both passive and active techniques. Passive techniques depend on contrast agents or susceptibility artifacts that enhance the appearance of the catheter in the image itself, whereas active techniques rely on supplemental hardware built into the catheter, such as a radiofrequency (RF) coil. Additionally, the ability to introduce an RF coil mounted on a catheter presents the opportunity to obtain high-resolution images of the vessel wall. These images can provide the capability to distinguish and identify various plaque components. The additional capabilities of MRI could potentially open up new applications within the purview of vascular interventions beyond those currently performed under X-ray fluoroscopic guidance. PMID- 11042635 TI - Intraoperative magnetic resonance imaging in epilepsy surgery. AB - The aim of this study was to investigate how intraoperative magnetic resonance imaging (MRI) can help in epilepsy surgery to asses immediately whether a resection or disconnection procedure is tailored to the individual needs of a patient, thus ideally meeting the treatment plan and enhancing the efficiency of the procedure. The recently proposed concept of an individually tailored procedure with as limited tissue removal as possible would support a more conservative resection than initially advocated by many centers; such limited removal would preserve as much brain as possible that is not necessarily epileptogenic or involved in propagation of seizures. For intraoperative imaging we used a Magnetom Open 0.2-T scanner located in our "twin-OR" in 61 patients with pharmacoresistant epilepsy. A three-dimensional sequence was used, allowing free slice reformatting. In the nonlesional cases (n = 32) the extent of the tailored temporal resection (n = 28) or callosotomy (n = 4) could be documented exactly. In the 29 lesional cases the complete resection was primarily proved in 23 patients. In three glioma patients a lesion that extended into eloquent areas did not allow for complete removal. A second look (n = 3) could increase the rate of total resection in the lesional cases from 79% to 90%. Intraoperative MRI allowed a reliable evaluation of the extent of resection or disconnection in epilepsy surgery within the operative procedure. It also provided the possibility of a second look in cases of incomplete resection, especially in the lesional cases. Increased knowledge of structure-function relationships as partially defined by intraoperative imaging may reduce the adverse neuropsychological sequelae of epilepsy surgery in the future. PMID- 11042636 TI - Nerve root infiltration of the first sacral root with MRI guidance. AB - The purpose of this clinical trial was to describe the methodology and evaluate the accuracy of optical tracking-based magnetic resonance (MR)-guided infiltration of the first sacral (S1) root. Thirty-five infiltrations were performed on 34 patients with a 0. 23-T open C-arm magnet installed in a fully equipped operation room with large-screen (36 inches) display and optical navigator utilizing infrared passive tracking. T1 and T2 fast spin-echo (FSE) images were used for localizing the target and fast field echo for monitoring the procedure. Saline as contrast agent in single-shot (SS)FSE images gave sufficient contrast-to-noise ratio. Twenty-four patients had unoperated L5/S1 disc herniation, and 10 had S1 root irritation after failed back surgery. Needle placement was successful in 97% of the cases, and no complications occurred. Outcome was evaluated 1-6 months (mean 2.2 months) after the procedure and was comparable to that of other studies using fluoroscopy or computed tomography guidance. MR-guided placement of the needle is an accurate technique for first sacral root infiltration. PMID- 11042637 TI - MRI-guided celiac plexus block. AB - Celiac plexus block is used as a palliative procedure in cases of severe upper abdominal pain caused by pancreatitis or tumors of the pancreas. It can be guided by bony landmarks, fluoroscopy, ultrasound (US), or computed tomography (CT). To avoid severe complications, methods visualizing soft tissue, like CT and magnetic resonance (MR) imaging, are preferable. We describe celiac plexus blocks carried out in an open MR scanner, offering needle guidance with an optical tracking system and near real-time image acquisition. Eight patients with severe chronic abdominal pain were included. In these, 14 celiac blocks were carried out. Good or total pain relief was achieved in 8 of the 14 blocks (57%), a moderate effect in 5 blocks (36%), and no effect in 1 block (7%). The placement of the needle was easily guided with MR in all cases. The MR technique ensures good visualization of soft tissue, direct monitoring of needle movement and avoids exposure to ionizing radiation. Celiac plexus block can safely be carried out in an open MR scanner. PMID- 11042638 TI - Magnetic resonance-guided percutaneous laser ablation of uterine fibroids. AB - Laser ablation of uterine fibroids using a percutaneous approach under local anesthetic in an open magnetic resonance (MR) scanner was performed in 12 symptomatic women awaiting hysterectomy. Accurate laser fiber placement was assisted by the use of an MR needle tracking system, as well as laser heat dissipation monitored during treatment by a real-time imaging processor. This day case procedure was well tolerated by all women, with eight women subsequently declining their planned surgery. Follow-up measurements of treated fibroid volume by MRI demonstrated a mean decrease of 37.5% at 3 months. This novel minimally invasive approach offers an alternative to surgery for women with fibroids, but longer follow-up is required to ascertain maximal fibroid shrinkage and to compare outcome with traditional surgery. PMID- 11042639 TI - Local hyperthermia with MR-guided focused ultrasound: spiral trajectory of the focal point optimized for temperature uniformity in the target region. AB - The objective of hyperthermia treatment is to deliver a similar therapeutic thermal dose throughout the target volume within a minimum amount of time. We describe a noninvasive approach to this goal based on magnetic resonance imaging (MRI)-guided focused ultrasound (FUS) with a spherical transducer that can be moved along two directions inside the bed of a clinical MR imager and that has an adjustable focal length in the third dimension. Absorption of FUS gives rise to a highly localized thermal buildup, which then spreads by heat diffusion and blood perfusion. A uniform temperature within a large target volume can be obtained using a double spiral trajectory of the transducer focal point together with constant and maximum FUS power. Differences between the real and target temperatures during the first spiral are evaluated in real time with temperature MRI and corrected for during the second spiral trajectory employing FUS focal point velocity modulation. Once a uniform temperature distribution is reached within the entire volume, FUS heating is applied only at the region's boundaries to maintain the raised temperature levels. Heat conduction, together with the design and timing of the trajectories, therefore ensures a similar thermal dose for the entire target region. Good agreement is obtained between theory and experimental results in vitro on gel phantoms, ex vivo on meat samples, and in vivo on rabbit thigh muscle. Edema in muscle was visible 1 hour after hyperthermia as a spatially uniform rise of the signal intensity in T(2)-weighted images. PMID- 11042640 TI - Thermal lesion conspicuity following interstitial radiofrequency thermal tumor ablation in humans: a comparison of STIR, turbo spin-echo T2-weighted, and contrast-enhanced T1-weighted MR images at 0.2 T. AB - The purpose of this study was to compare the contrast between radiofrequency (RF) thermal liver lesions and surrounding tissue in T2-weighted turbo spin-echo sequences (TSE T2), short TI inversion recovery techniques (STIR), and contrast enhanced (CE) T1-weighted spin-echo images. Nineteen RF thermal ablations were performed on eight patients with metastatic liver tumors. After ablation, contrast-to-noise ratios (CNRs) were calculated between mean signal amplitudes from three regions of interest (ROI) (lesion, surrounding edema, and normal tissue) using TSE T2-weighted, STIR, and contrast-enhanced T1-weighted (CE T1) sequences for each lesion. CNRs between the thermal lesion and normal liver tissue for both TSE T2-weighted (mean 0.9) and STIR (2.0) images were significantly lower than for CE T1-weighted (8.4) images (t-test, alpha = 0.05). However, CNRs between edema rim and the core of the thermal lesion for both TSE T2-weighted (8.1) and STIR images (7.2) were not significantly different (t-test, alpha = 0.05) from CNRs between lesion and normal tissue for CE T1-weighted images (8.4), nor was the CNR between edema rim and normal tissue for both TSE T2 weighted (10.3) and STIR (9.8) images. Although the edema was not visible on CE T1-weighted images, 18 of 19 lesions (94.7%) were surrounded by a hyperintense rim on TSE T2-weighted or STIR images. Both TSE T2-weighted and STIR sequences represent valid techniques for repeatable assessment of RF thermal lesions. PMID- 11042641 TI - Toward MRI-guided coronary catheterization: visualization of guiding catheters, guidewires, and anatomy in real time. AB - The success of x-ray fluoroscopy-guided coronary catheterization depends in part on the ability to obtain simultaneous and real-time visualization of the guidewire, guiding catheter, and anatomy of the chest. The hypothesis explored in this paper is that magnetic resonance imaging (MRI) could provide this ability. This hypothesis was tested with loopless antennas used as the guidewire and a guiding catheter and two surface coils, each connected to four different receiver channels of a GE 1.5-T CV/I MRI scanner. Experiments were conducted on six healthy dogs. Intravascular antennas were inserted in the right carotid artery and maneuvered in the aorta while running a fast gradient-echo sequence (TR/TE 5/1.3 msec, flip angle 7 degrees). Real-time projection images of the chest anatomy, together with the guidewire and guiding catheter, were obtained. Positioning of the MRI guiding catheter either in the descending aorta, ascending aorta, or heart was achieved easily. This study represents a step toward MRI guided coronary catheterization. PMID- 11042642 TI - The use of carbon dioxide in magnetic resonance angiography: a new type of black blood imaging. AB - The injection of boluses of carbon dioxide (CO(2)) during T1-weighted magnetic resonance angiography (MRA) resulted in sharp depiction of the vascular lumen during the performance of in vitro and in vivo animal experiments. We propose the use of the low-cost, relatively safe CO(2) as a negative contrast agent for MRA and speculate on its use in future interventional MRA. PMID- 11042643 TI - Placement of an inferior vena cava filter in a pig guided by high-resolution MR fluoroscopy at 1.5 T. AB - Percutaneous placement of an inferior vena cava filter is a means for long-term prevention of pulmonary thromboembolism. In this study we investigated the magnetic resonance (MR) imaging properties of a Nitinol vena cava filter, in various anatomic and angiographic scans, as well as the feasibility of placing this filter under near real-time, high-resolution MR fluoroscopy. We made use of the passive tracking strategy, with on-line image processing and visualization, both in vitro and in a pig. The artifacts provoked by the metallic filter were such that the position and orientation of the filter were well depicted in all scans. Considerable radiofrequency caging obscured the interior of the filter. Our experiments showed that an MR-guided vena cava filter placement, with sufficient temporal and spatial resolution, is possible. Three-dimensional phase contrast MRA allowed direct evaluation of the filter placement procedure, without the use of contrast agent. PMID- 11042644 TI - MR imaging in the presence of vascular stents: A systematic assessment of artifacts for various stent orientations, sequence types, and field strengths. AB - A systematic evaluation of the potential quality of magnetic resonance images recorded in the presence of metallic stents was performed on a low-field open imager operating at 0.2 T and on a high-field closed unit operating at 1.0 T. Eight different stent types were examined by two-dimensional gradient-echo sequences with echo times of 4 and 10 msec and by a fast spin-echo technique. In addition, a three-dimensional gradient-echo sequence was applied with an echo time of 2.4 msec. A set of sequence and slice parameters was used on both scanners. Thus, artifacts due to susceptibility effects depending on the magnetic field strength could be distinguished from radiofrequency shielding effects in the lumen of the stents (independent of the field strength). Nine different orthogonal orientations of the stent axis and the image (in terms of slice, read, and phase-encoding direction) were tested, and the artifacts (extension of signal void and visibility of the lumen) were compared. The optimal strategy for visualization of vascular and perivascular regions outside the stents was fast spin-echo imaging with the stent axis and read direction parallel to the static field. Susceptibility-induced signal void in gradient-echo images was minimal using the three-dimensional approach. Increased transmitter amplitudes above usual values provided clearly improved insight in the lumen using gradient-echo sequences. PMID- 11042645 TI - Real-time MR fluoroscopy for MR-guided iliac artery stent placement. AB - The purpose of this study was to test the feasibility of real-time magnetic resonance (MR) guidance of iliac artery stent placement. Radial scanning together with the sliding window reconstruction technique was implemented on a 1.5 T magnet, yielding a frame rate of 20 images per second. Seven prototype nitinol ZA stents were deployed in iliac arteries of living pigs under MR control. All stents were well visualized on the radial MR images, allowing depiction of the mounted stents as well as stent deployment without anatomy-obscuring artifacts. Stent placement was sucessful in all cases and took 6 minutes on average. The position of the stents was correctly visualized by real-time radial MR scanning, as proved by digital subtraction X-ray angiography. Combined radial scanning and the sliding window reconstruction technique allow real-time MR-guided stent placement in iliac arteries. PMID- 11042646 TI - On-line flow quantification by low-resolution phase-contrast MR imaging and model based postprocessing. AB - Over the past decade, magnetic resonance (MR) imaging has been developed toward a tool for guiding and evaluating diagnostic and therapeutic interventions. Within the field of vascular MR-guided interventions, MR has potential for providing on line monitoring of the blood volume flow rate, which is relevant during procedures such as balloon angioplasty and stent placement. We recently reported a hardware and software environment for enabling flow quantification every 8 seconds using nontriggered phase-contrast imaging. In the present study, the objective was to increase temporal resolution further to one evaluation per 4 seconds. We achieve this by lowering spatial resolution to 3 pixels per lumen diameter. The accuracy of the measurements is preserved by applying model-based postprocessing for quantification of the volume flow rate. Phantom and volunteer studies are presented, demonstrating the accuracy of the model-driven approach for the applied short acquisitions. The capabilities of the presented approach are illustrated by the results of several hypercapnia experiments and carotid compression tests performed on healthy volunteers. PMID- 11042647 TI - An optical system for wireless detuning of parallel resonant circuits. AB - A new optical method of detuning parallel resonant circuits is described. This method involves the integration of a photoresistor in parallel with the inductor and capacitor of a parallel resonant circuit, in this case a magnetic resonance imaging (MRI) receiver coil. A fiberoptic cable extending the length of the interventional device is used in conjunction with an external light source to deliver light to the photoresistor. Exposing the photoresistor to light changes its bulk resistance and greatly lowers the Q of the parallel resonant circuit, effectively detuning it. By combining this optical detuning scheme with inductive coupling of the interventional device-mounted microcoils to a standard MRI coil, a completely wireless device for active device tracking has been created. This new device improves on current technology by simplifying device complexity and reducing patient risk by eliminating the need for electrical connections between the device-mounted microcoils to the MR receiver channel. PMID- 11042648 TI - MR-guided balloon angioplasty of stenosed aorta: in vivo evaluation using near standard instruments and a passive tracking technique. AB - The purpose of this study was to assess the feasibility of magnetic resonance (MR)-guided balloon angioplasty of a stenosed aorta on an open low-field magnet using a passive tracking technique. Visualization of vessels and position of instruments were realized by using a fast low-angle shot (FLASH) sequence. Catheters and guidewire were prepared for susceptibility-based MR visualization. Standard balloon catheters were inflated with diluted gadolinium, and nitinol guidewires were modified by incorporation of iron oxide markers into their walls. After validation on a flow phantom, balloon angioplasty was performed on an in vivo model of arterial stenosis. Creation of abdominal aorta stenosis was realized in five piglets. MR-guided balloon angioplasty of the aorta was performed with success in all but one. In one of them, stent implantation was achieved in the descending aorta. Balloon angioplasty using a passive tracking technique is a simple concept that can be realized with near-standard instruments and any MR imaging system. This represents an advance toward MR-guided vascular interventions in the future. PMID- 11042649 TI - Accurate localization of needle entry point in interventional MRI. AB - In interventional magnetic resonance imaging (MRI), the systems designed to help the surgeon during biopsy must provide accurate knowledge of the positions of the target and also the entry point of the needle on the skin of the patient. In some cases, this needle entry point can be outside the B(0) homogeneity area, where the distortions may be larger than a few millimeters. In that case, major correction for geometric deformation must be performed. Moreover, the use of markers to highlight the needle entry point is inaccurate. The aim of this study was to establish a three-dimensional coordinate correction according to the position of the entry point of the needle. We also describe a 2-degree of freedom electromechanical device that is used to determine the needle entry point on the patient's skin with a laser spot. PMID- 11042650 TI - Dose-dependent effect of etoposide in combination with busulfan plus cyclophosphamide as conditioning for stem cell transplantation in patients with acute myeloid leukemia. AB - To evaluate the efficacy and toxicity of two different etoposide (VP-16) dosages (30 or 45 mg/kg) in combination with busulfan/cyclophosphamide as conditioning therapy followed by stem cell transplantation in acute myeloid leukemia (AML), 90 patients with AML received either 30 mg/kg (n = 60) or 45 mg/kg (n = 30) etoposide in combination with busulfan (16 mg/kg) and cyclophosphamide (120 mg/kg). The stem cell source was allogeneic related bone marrow (BM) (n = 53), allogeneic unrelated BM (n = 5), allogeneic unrelated peripheral blood (PBSC) (n = 2), syngeneic BM (n = 2), autologous BM purged (n = 9) or unpurged (n = 9), autologous PBSC (n = 10). Fifty-six patients (62%) were in first CR, 26 (29%) were > first CR, and eight (9%) were transplanted in relapse. Principal toxicities in both groups were mucositis and hepatotoxicity. Forty-five mg/kg etoposide resulted in greater hepatic toxicity (P = 0.03), and a higher incidence of VOD (23 vs 12%, P = 0.04) and acute GVHD grade III/IV (13 vs 5%, NS). The treatment-related mortality was 17% in the 30 mg/kg group and 33% in the 45 mg/kg group, mainly due to infections, intestinal pneumonia and GVHD. Hematological recovery of leukocytes 1/nl was comparable in both groups (17 vs 16 days). After a median follow-up of 16 months 19% in the 30 mg/kg group and 23% in the 45 mg/kg group relapsed. In patients who had undergone allogeneic related bone marrow transplantation in first CR no relapses occurred after a median follow-up of 3 years. For all patients the 3-year estimated disease-free survival was 62% in the 30 mg/kg group and 40% in the 45 mg/kg group (P = 0.03). For patients in first CR who underwent allogeneic related stem cell transplantation the 3 year disease free survivals were 80% and 66%, respectively (P = 0.4). We conclude that etoposide 30 mg/kg or 45 mg/kg in combination with busulfan/cyclophosphamide is a highly active regimen for bone marrow transplantation of patients with AML with a low relapse rate. However, conditioning with 30 mg/kg rather than 45 mg/kg etoposide resulted in less toxicity and a better overall survival due to a lower transplant-related mortality. Bone Marrow Transplantation (2000) 26, 711-716. PMID- 11042651 TI - A comparative study of sequential priming and mobilisation of progenitor cells with rhG-CSF alone and high-dose cyclophosphamide plus rhG-CSF. AB - Stem cell mobilisation can be achieved either by administration of rhG-CSF alone or after high-dose cyclophosphamide (HDCy) plus rhG-CSF. We have compared both mobilisation procedures intra-individually in 43 patients with haematological malignancies. Furthermore, the toxicity data were registered. The CD34+ cell yield was higher after mobilisation with HDCy plus rhG-CSF than after rhG-CSF alone in 21 out of 22 patients who were actually harvested after both procedures. If a patient mobilised insufficiently after rhG-CSF alone, the yield of CD34+ cells after the following HDCy priming was lower compared to patients who mobilised sufficiently after rhG-CSF priming alone. In 12 patients with B cell malignancies a reduced number of B cells such as CD10+, CD19+, CD20+ cells in bone marrow as well as in leukapheresis products was observed after HDCy plus rhG CSF compared to rhG-CSF alone. Toxicity data revealed HDCy as a relatively toxic priming regimen with all patients hospitalised and 74% experiencing neutropenic fever and administration of intravenous antibiotics. In two patients, seizure like episodes were observed during cyclophosphamide bolus infusion. In conclusion, HDCy increased the yield of CD34+cell and reduced B cells in leukapheresis products indicating reduced tumour cell load compared with rhG-CSF priming alone. The efficacy of HDCy priming is limited by its profound toxicity and morbidity. Studies evaluating efficacy and safety of lower doses of cyclophosphamide are needed. Bone Marrow Transplantation (2000) 26, 717-722. PMID- 11042652 TI - Matched-pair analysis of peripheral blood stem cells compared to marrow for allogeneic transplantation. AB - We performed a case-control analysis of 42 patients with advanced leukemia or MDS comparing peripheral blood stem cell (PBSC) with marrow grafts (BMT) from HLA matched sibling donors. PBSC were mobilized with G-CSF (7.5 microg/kg/day) and yielded a median of 6.7 x 10(6) CD34+ cells/kg (range, 1.6-15.0) and 2.7 x 10(8) CD3+ cells/kg (range, 1.1-7.1) vs marrow grafts with a median of 2.0 x 10(8) nucleated cells/kg (range, 1.8-2.2). Recovery was significantly faster after PBSCT compared to BMT, with a median of 17 (range, 12-26) vs 26 (range, 16-36) days, respectively, to neutrophils >0.5 x 10(9)/l (P < 0.01), and 22 (range, 12 >60) vs 42 (range, 18->60) days, for platelet recovery (P < 0.01). Transplantation of >/=7 x 10(6) CD34+ cells/kg accelerated recovery to >20 x 10(9) l platelets; median 17 days (range, 12-19) vs 23 days (range, 17-36) for those receiving <7 x 10(6)/kg (P = 0.01). PBSC and marrow recipients had similar risks of grades II-IV or III-IV acute GVHD or extensive chronic GVHD (all P > 0.3). At 1 year after PBSCT and BMT, the risk of relapse was 41% and 32%, respectively (P = 0.47), and the probability of survival was 46% and 48%, respectively (P = 0.70). HLA-matched sibling PBSCT resulted in faster neutrophil and platelet engraftment compared to BMT, with no subsequent differences in acute or chronic GVHD, relapse or survival. A minimum of 7 x 10(6) CD34+ cells/kg in PBSC grafts may be required for very rapid platelet engraftment. Bone Marrow Transplantation (2000) 26, 723-728. PMID- 11042653 TI - Use of the anti-idiotype antibody vaccine TriAb after autologous stem cell transplantation in patients with metastatic breast cancer. AB - Between April 1997 and March 1998 we evaluated the immune response and outcome in 11 chemosensitive patients who were treated with the anti-idiotype antibody vaccine TriAb after recovery from intensive therapy and autologous stem cell transplant (ASCT). Triab was commenced after recovery from the acute effects of ASCT; a minimum interval of 1 month was required from completion of consolidation radiotherapy, if given. Nine patients (82%) manifest anti-anti-idiotype antibody (Ab3) responses post ASCT. The maximal Ab3 response was seen after a median of 10 doses (range 5-20), which corresponded to a median of 14 months (range 5-19) post ASCT. Evidence of a T cell proliferative response was seen in eight patients; the response was modest in most of these. At a median follow-up of 24 months (range 22-33) after ASCT, four patients are alive without evidence of disease progression. All four of these patients were in the subgroup with more vigorous immune responses. Subsequent efforts have been directed toward the achievement of higher levels of immune responses more rapidly post ASCT. Bone Marrow Transplantation (2000) 26, 729-735. PMID- 11042654 TI - Feasibility and toxicity of high-dose chemotherapy supported by peripheral blood stem cell transplantation in elderly patients (>/=60 years) with non-Hodgkin's lymphoma: comparison with patients <60 years treated within the same protocol. AB - Limited data are available concerning feasibility and toxicity of progenitor cell mobilization and high-dose therapy (HDT) supported by peripheral blood stem cell transplantation (PBSCT) in elderly patients (>/=60 years) with non-Hodgkin's lymphoma (NHL). From 1995 to 1999, 17 elderly NHL patients (median age 63 years, range 60-70) entered our HDT program and were mobilized with CY (4 g/m2) followed by G-CSF. Mobilization was successful in 13 patients, who then received BEAM or BEAC followed by PBSCT. The feasibility and toxicity of progenitor cell mobilization and HDT in the elderly patients were compared with experiences in 62 NHL patients <60 years (median 46 years, range 16-59), who received the same mobilization protocol and of whom 48 patients received HDT supported by PBSCT. No significant differences were observed between these groups in the success rate of progenitor cell mobilization, in the number of CD34-positive cells collected or in the number of aphereses needed. HDT appeared to be somewhat more toxic in the elderly patients: a higher peak CRP value (P = 0.08) and longer in-hospital stay (P = 0. 05) were observed. No differences were found in transplant-related mortality or severe organ toxicity between these age groups except for oral mucositis grade >2, which tended to be more common in the elderly patients (P = 0.07). We conclude that progenitor cell mobilization and HDT supported by PBSCT is also feasible in selected elderly patients with NHL. Bone Marrow Transplantation (2000) 26, 737-741. PMID- 11042655 TI - Lymphocyte subset reconstitution after HLA-identical placental blood transplantation (PBT) or PBT plus bone marrow transplantation (BMT) in three children with beta-thalassemia major. AB - The kinetics of circulating lymphoid cells were evaluated in three children suffering from beta-thalassemia major after HLA-identical sibling placental blood transplant (PBT) in one patient and placental blood plus bone marrow transplantation (BMT) in two patients. Recovery of the main lymphocyte subsets, as determined by phenotype analysis of circulating PBMCs, was complete within 2 months after transplant. NK (CD56+) cells were the first to appear in peripheral blood, followed by T (CD3+, CD2+, CD7+) and B (CD19+) cells. Of the T lymphocytes, the CD8+ were the first to reconstitute, but recovery of CD4+ cells was also rapid and within 6 months these T cells reached normal values. The expression of CD57 by NK or T cells was slightly delayed. The evaluation of RA and RO isoform expression of the CD45 molecule showed a prevalence of the CD45RA antigen with a ratio of 2-3:1. In the PBT only patient, T cells expressing the CD45RO antigen prevailed in the early post-transplant period. Severe or chronic GVHD was not observed. This experience demonstrates that reconstitution of lymphocyte subsets is successful in genetic hematological diseases after transplantation of HLA-identical placental blood or placental blood plus bone marrow from healthy or heterozygous siblings. Bone Marrow Transplantation (2000) 26, 743-747. PMID- 11042656 TI - Transplantation of ABO-incompatible bone marrow and peripheral blood stem cell components. AB - Hemolysis may occur during infusion of an ABO-incompatible HSC component if the recipient has isoagglutinins directed against donor red blood cells, or later as a result of the production by donor lymphocytes of isoagglutinins directed against recipient ABO-antigens. Peripheral blood stem cell (PBSC) components collected by apheresis contain few red cells but considerably greater numbers of lymphocytes than marrow. We reviewed the transplant courses of 158 recipients of marrow (n = 90) or PBSC (n = 68) from HLA-identical, ABO-incompatible sibling donors. No patient experienced immediate or delayed hemolysis attributable to the ABO incompatibility. Recipients of minor ABO-incompatible red cell-replete marrow required fewer red cell transfusions during the first week after transplantation than recipients of PBSC or marrows depleted of red cells; the red cell transfusion requirements for the following 3 weeks did not differ. The maximum level of bilirubin did not differ for patients classified by ABO incompatibility or source of HSC. The development of positive antiglobulin tests occurred for eight marrow recipients from a separate group of 22 patients (17 marrow, five PBSC) for whom this testing was performed. None of these patients developed overt hemolysis. These data indicate that hemolysis complicating ABO-incompatible transplantation is not common after either marrow or PBSC transplantation. Bone Marrow Transplantation (2000) 26, 749-757. PMID- 11042657 TI - Treatment and outcome of invasive Aspergillus infections in allogeneic BMT recipients. AB - The outcome of invasive aspergillosis (IA) has been considered poor in allogeneic BMT recipients. We analyzed retrospectively the treatment and outcome of IA diagnosed during life in a recent cohort of 20 allogeneic BMT recipients. All patients were initially treated with amphotericin B (AmB) (conventional 16, liposomal 4). Due to toxicity, conventional AmB was changed to a liposomal preparation in 10 patients. Five patients also received itraconazole and three underwent surgery. Of 19 evaluable patients, two patients achieved a complete response and a partial response was observed in five patients (response rate 37%). The median survival was 37 days after the diagnosis. Only two patients (10%) were cured. The prognosis of allogeneic BMT recipients with IA has remained poor. Although treatment responses are common, immunosuppression aggravated by GVHD and its treatment, as well as the commonly disseminated presentation of IA, seem to be major obstacles to the success of therapy. Bone Marrow Transplantation (2000) 26, 759-762. PMID- 11042658 TI - A survey of allogeneic bone marrow transplant programs in the United States regarding cytomegalovirus prophylaxis and pre-emptive therapy. AB - Despite an extensive literature, no consensus has emerged regarding the optimal preventive strategy for CMV in allogeneic bone marrow transplantation (BMT). No survey of CMV prevention in BMT centers in the United States has yet been published. A questionnaire was sent to all allogeneic BMT programs in the United States, as listed in the November 1998 National Marrow Donor Program (NMDP) address roster. Questions included whether universal prophylaxis, pre-emptive therapy, or some other strategy was used for CMV prevention, and which CMV diagnostic tests were utilized. Eighty-one of 96 programs (86%) responded to the survey. Of these, 46 (56%) utilize a pre-emptive ganciclovir strategy, whereas 17 (21%) utilize universal prophylaxis, and 15 (19%) utilize a hybrid strategy based on risk stratification. The most commonly utilized CMV diagnostic tests are CMV DNA by PCR (55 centers), shell vial centrifugation culture (52), tissue culture (42), pp65 antigenemia assay (38), and CMV-DNA by Digene hybrid capture (14). Of these, the CMV-DNA by PCR, pp65 antigenemia assay, and shell vial culture are the most frequently utilized as triggers for pre-emptive therapy. Quantitative assays are common (PCR 42%, Digene 64%). We conclude that centers currently performing allogeneic BMT in the United States employ a variety of strategies for CMV prevention, and differ in their diagnostic tests of choice for CMV monitoring. These results emphasize the need for large-scale studies to identify optimal diagnostic and management protocols. Bone Marrow Transplantation (2000) 26, 763 767. PMID- 11042659 TI - Donor leukocyte infusion for Japanese patients with relapsed leukemia after allogeneic bone marrow transplantation: lower incidence of acute graft-versus host disease and improved outcome. AB - To clarify the role of donor leukocyte infusion (DLI) in the treatment of leukemia relapsing after allo-BMT, data from 100 patients were collected from 46 facilities in Japan and analyzed with respect to the efficacy and adverse effects of donor leukocyte infusion. Complete remission was achieved in 11 of 12 (91%) patients with relapsed chronic myelogenous leukemia (CML) in chronic phase, three of 11 (27%) with CML in the acute phase, eight of 21 (38%) with acute myelogenous leukemia (AML), six of 23 (25%) with acute lymphoblastic leukemia (ALL) and five of 11 (45%) with myelodysplastic syndrome (MDS). The probability of remaining in CR at 3 years was 82% in CML patients in the chronic phase, but 0% in those with CML in the acute phase, 7% in those with AML, 0% with ALL and 33% with MDS. Acute GVHD (>/=2) developed in 31 of 89 (34%) patients with HLA-identical related donors and was fatal for seven (7%). Cytopenia developed in 21 of 94 (22%) with no associated fatalities. When the outcome of patients with CML in CP and MDS was analyzed, development of GVHD, cytopenia, or both, was associated with a higher GVL effect (15 of 16, 93%) than in those without adverse affects (one of 6, 17%). A leukocyte dose of 5 x 107/kg of recipient body weight appeared to be optimal as an initial dose of DLI. Given the relatively low incidence of acute GVHD and the similar GVL effect, DLI may be more beneficial to patients in Japan with recurrent leukemia than to those in Western countries. Bone Marrow Transplantation (2000) 26, 769-774. PMID- 11042660 TI - Scintigraphic study of the major salivary glands in pediatric bone marrow transplant recipients. AB - Total body irradiation (TBI) at bone marrow transplantation (BMT) is shown to cause salivary gland dysfunction in children. The aim of the investigation was to study the function of major salivary glands in long-term surviving children following treatment with TBI, using salivary gland scintigraphy (SGS). Thirteen patients (seven male, six female), who had received TBI before the age of 13 years and survived more than 4 years, participated in the study. A reference group of 10 patients (nine male, one female) was examined shortly before they were to undergo BMT. The mean age was 14.1 +/- 4.1 years in the TBI-treated group and 12.8 +/- 5.9 years in the reference group. Unstimulated and stimulated whole salivary secretion rates were measured for 15 and 5 min, respectively, before SGS was performed. The percentage of stimulated secretion was 44.7 +/- 18.1% in the TBI-treated group compared to 58.4 +/- 13.0% in the reference group (P = 0.0438). Slower reaccumulation after excretion was found in the TBI-treated patients compared to the reference group (P = 0. 0300). The function of the major salivary glands in long-term survivors treated with TBI at BMT before the age of 13 years was found to be diminished, as shown by the reduced trapping rate and reduced emptying capacity, compared to prior to BMT. Bone Marrow Transplantation (2000) 26, 775-779. PMID- 11042661 TI - Permanent tunneled silicone central venous catheters for autologous PBPC harvest in children and young adults. AB - Children with high risk malignancies are usually given permanent (Hickman-type) tunneled silicone rubber central venous catheters (silicone CVCs) for the administration of chemotherapy. In the past, these children received an additional short-term polyurethane dialysis CVC for stem cell apheresis. To avoid placement of an additional short-term CVC, we started in 1995 to use pre-existing silicone CVCs for PBPC harvests. From May 1996 to February 1999 we evaluated 165 harvests in 37 children and 14 young adults (16-28 years) treated with high-dose chemotherapy and stem cell support, comparing CD34+ cell harvest efficiency, catheter tolerability, and complications in three different approaches to vascular access. Pre-existing silicone CVCs (64%) or peripheral venous cannulae (15%) were the first choice for venous access. Only when these failed were polyurethane CVCs (21%) used. No significant difference was seen between these three groups, even after dividing the silicone CVC group (105 harvests in 32 patients) into three subgroups according to weight and age. The most frequent problems were citrate toxicity (n = 33), mechanical obstruction inside (n = 9) and outside the cell separator (n = 2), decreased draw line flow in silicone CVCs (n = 7), decreased draw line flow in peripheral venous cannulae (n = 6), and one occlusion in a polyurethane CVC. Pre-existing CVCs and peripheral venous cannulae functioned efficiently when used as a draw line in 79% of the apheresis procedures without significantly reducing single harvest efficiency or catheter tolerability. Consequently, the risks and costs associated with the placement of a dialysis CVC could be avoided in the majority of cases. Bone Marrow Transplantation (2000) 26, 781-786. PMID- 11042662 TI - Large scale purification of human blood CD34+ cells from cryopreserved peripheral blood stem cells, using a nylon-fiber syringe system and immunomagnetic microspheres. AB - Isolation of large numbers of human peripheral blood CD34+ cells could lead to therapeutic applications, including purging of malignant cells from blood cell transplantations, purging of T cells from allogeneic bone marrow, and even blood cell transplantation. This procedure has limitations if there are not sufficient numbers of progenitor cells in the leukapheresis concentrates available for selection after detection of tumor cells in apheresis products. Use of frozen/thawed peripheral blood mononuclear cell (PBMC) samples would make feasible pooling of two or even more stem cell harvests collected at different time points and the total number of CD34+ progenitor cells available would increase. We established an efficient method for purification of CD34+ cells from cryopreserved apheresis products, using a nylon-fiber syringe system and immunomagnetic microspheres. We compared purity, recovery rate and clonogenicity of CD34+ cells purified from fresh (n = 22) and cryopreserved apheresis products (n = 14), using a nylon-fiber syringe system and immunomagnetic microspheres. The purity of CD34+ cells from cryopreserved products was less than that from fresh products (85.9 +/- 14.4% vs 94.6 +/- 10.0%), but the recovery rate of CD34+ cells and colony-forming cells was comparable between fresh and cryopreserved products. One patient underwent grafting with peripheral blood CD34+ cells selected after freezing, with good success. Therefore, these cells are capable of rapidly reconstituting hematopoiesis after high-dose chemotherapy. Bone Marrow Transplantation (2000) 26, 787-793. PMID- 11042663 TI - Varicella zoster meningoencephalitis following treatment for dermatomal zoster in an alloBMT patient. AB - Herpes zoster infections are frequently observed after allogeneic bone marrow transplantation (alloBMT). In the majority of cases, the infection is restricted to specific dermatomes and responds to oral acyclovir, without visceral dissemination. We report the case of a 40-year-old male who developed dermatomal herpetic infection 8 months post alloBMT. The herpetic rash responded well to treatment with high-dose oral acyclovir. However, within a week of cessation of therapy, the patient re-presented with dermatomal zoster and meningoencephalitis. Although the cutaneous lesions resolved with intravenous acyclovir, clinical features of meningoencephalitis persisted, along with evidence of varicella zoster virus (VZV) DNA in cerebrospinal fluid (CSF). A satisfactory response to treatment was observed only after the addition of intravenous foscarnet to acyclovir. Based on our experience with this patient, we suggest that in a subset of alloBMT recipients, late dermatomal herpes zoster infections may respond only partially to treatment with standard oral acyclovir. The use of oral acyclovir preparations with higher bioavailability (valacyclovir) or intravenous acyclovir early on may prevent the considerable morbidity associated with disseminated zoster infection. Bone Marrow Transplantation (2000) 26, 795-796. PMID- 11042664 TI - Reversible leukoencephalopathy associated with re-infusion of DMSO preserved stem cells. AB - We report a case of posterior reversible leuko- encephalopathy (PRL) following the infusion of dimethylsulfoxide (DMSO) cryopreserved autologous stem cells in the setting of myeloablative chemotherapy in a patient with recurrent Ewing's sarcoma. Magnetic resonance (MR) imaging revealed white matter changes which resolved over the next 2 months. Bone Marrow Transplantation (2000) 26, 797-800. PMID- 11042665 TI - Cyclosporin A-related cerebral vasculopathy. AB - The use of cyclosporin A has been associated with several side-effects, including neurotoxicity. The mechanism of toxicity is not well known. We report two patients treated with cyclosporin A who developed lesions in the cerebral white matter associated with abnormally elevated cerebral blood flow velocities on transcranial doppler ultrasound and abnormal vascular appearance on magnetic resonance angiography. Bone Marrow Transplantation (2000) 26, 801-804. PMID- 11042666 TI - Allogeneic peripheral blood stem cell transplantation for the treatment of chronic active Epstein-Barr virus infection. AB - The prognosis of chronic active Epstein-Barr virus infection (CAEBV) is very poor. We describe a 24-year-old male with severe CAEBV who was treated with allogeneic peripheral blood stem cell transplantation (allo-PBSCT). On admission, EBER-1 in lymphocytes infiltrating the liver, EBV-DNA in peripheral blood mononuclear cells (PBMC) and monoclonal NK cell proliferation were confirmed. After unsuccessful chemotherapy, he received an allo-PBSCT from his HLA-identical sister. Although he died of pulmonary hemorrhage on day +19, EBV-DNA was undetectable by PCR in PBMC, and the post-mortem liver showed no EBER-1-positive lymphocytes. This experience suggests that EBV-positive lymphocytes in CAEBV may be eradicated by allo-PBSCT, thereby raising the possibility of a new treatment modality. Bone Marrow Transplantation (2000) 26, 805-808. PMID- 11042667 TI - Long-term molecular remission induced by donor lymphocyte infusions for recurrent acute myeloblastic leukemia after allogeneic bone marrow transplantation. AB - A case of acute myelogenous leukemia with a t(8;21) translocation relapsed 5 months after allogeneic bone marrow transplantation (allo-BMT). After chemotherapy-induced hematologic remission, the patient received donor lymphocyte infusions (DLI); 4.9 x 108/kg T cells were infused. After DLI, she achieved molecular CR for the first time after allo-BMT, which lasted for 40 months. However, she suffered from grade III acute GVHD of the skin and the liver. Hepatic GVHD was sustained and resulted in fatal outcome. The case demonstrates that DLI is a double-edged sword. Further study is necessary before DLI can be considered to be a beneficial therapy for acute leukemia. Bone Marrow Transplantation (2000) 26, 809-810. PMID- 11042668 TI - Unusual adverse events following peripheral blood stem cell (PBSC) mobilisation using granulocyte colony stimulating factor (G-CSF) in healthy donors. AB - Here, we describe two cases of severe pyogenic infection in healthy donors diagnosed immediately following stem cell mobilisation with G-CSF. In the first donor a painful perianal abscess and in the second one an apical abscess required surgical incision. The reported serious adverse events in the literature are reviewed and the potential pathophysiological role of G-CSF or GM-CSF in augmenting inflammatory processes is discussed. In the light of a rapidly increasing number of related and unrelated peripheral blood stem cell donations the need for more comprehensive donor work-up and follow-up for peripheral blood stem cell donors has to be considered. Bone Marrow Transplantation (2000) 26, 811 813. PMID- 11042669 TI - Trimethoprim/sulfamethoxazole prophylaxis impairs function of mobilised autologous peripheral blood stem cells. PMID- 11042670 TI - The TRAIL DISCussion: It is FADD and caspase-8! PMID- 11042671 TI - Ordering of ceramide formation, caspase activation, and Bax/Bcl-2 expression during etoposide-induced apoptosis in C6 glioma cells. AB - Etoposide (VP-16) a topoisomerase II inhibitor induces apoptosis of tumor cells. The present study was designed to elucidate the mechanisms of etoposide-induced apoptosis in C6 glioma cells. Etoposide induced increased formation of ceramide from sphingomyelin and release of mitochondrial cytochrome c followed by activation of caspase-9 and caspase-3, but not caspase-1. In addition, exposure of cells to etoposide resulted in decreased expression of Bcl-2 with reciprocal increase in Bax protein. z-VAD.FMK, a broad spectrum caspase inhibitor, failed to suppress the etoposide-induced ceramide formation and change of the Bax/Bcl-2 ratio, although it did inhibit etoposide-induced death of C6 cells. Reduced glutathione or N-acetylcysteine, which could reduce ceramide formation by inhibiting sphingomyelinase activity, prevented C6 cells from etoposide-induced apoptosis through blockage of caspase-3 activation and change of the Bax/Bcl-2 ratio. In contrast, the increase in ceramide level by an inhibitor of ceramide glucosyltransferase-1, D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol caused elevation of the Bax/Bcl-2 ratio and potentiation of caspase-3 activation, thereby resulting in enhancement of etoposide-induced apoptosis. Furthermore, cell-permeable exogenous ceramides (C2- and C6-ceramide) induced downregulation of Bcl-2, leading to an increase in the Bax/Bcl-2 ratio and subsequent activation of caspases-9 and -3. Taken together, these results suggest that ceramide may function as a mediator of etoposide-induced apoptosis of C6 glioma cells, which induces increase in the Bax/Bcl-2 ratio followed by release of cytochrome c leading to caspases-9 and -3 activation. PMID- 11042672 TI - Phenylephrine protects neonatal rat cardiomyocytes from hypoxia and serum deprivation-induced apoptosis. AB - Previous studies have shown that alpha-adrenergic activation reduces myocardial damages caused by ischemia/reperfusion. However, the molecular mechanisms of how alpha-adrenergic activation protects the myocardium are not completely understood. The objective of this study was to test the hypothesis that alpha adrenergic activation protects the myocardium by, at least in part, inhibiting apoptosis in cardiomyocytes. The current data has shown that apoptosis in neonatal rat cardiomyocytes, induced by 24 h treatment with hypoxia (95% N2 and 5% CO2) and serum deprivation, was inhibited by co-treatment with phenylephrine. Pre-treatment with phenylephrine for 24 h also protected cardiomyocytes against subsequent 24 h treatment with hypoxia and serum deprivation. Exposure of cardiomyocytes to phenylephrine for up to 9 days under normoxic conditions did not cause apoptosis. The phenylephrine-mediated cytoprotection was blocked by an alpha-adrenergic antagonist, phentolamine. beta-adrenergic activation with isoproterenol did not protect cardiomyocytes against hypoxia and serum deprivation-induced apoptosis. Under hypoxic conditions, phenylephrine prevented the down-regulation of Bcl-2 and Bcl-X mRNA/protein and induced hypertrophic growth. Phenylephrine-mediated protection was abrogated by the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor wortmannin and was mimicked by the caspase-9 peptidic inhibitor LEHD-fmk. These results suggest that alpha adrenergic activation protects cardiomyocytes against hypoxia and serum deprivation-induced apoptosis through regulating the expression of mitochondrion associated apoptosis regulatory genes, preventing activation of mitochondrial damage-induced apoptosis pathway (cytochrome C-caspase-9), and activating hypertrophic growth. PMID- 11042673 TI - CD40 ligand, Bcl-2, and Bcl-xL spare group I Burkitt lymphoma cells from CD77 directed killing via Verotoxin-1 B chain but fail to protect against the holotoxin. AB - Owing to its lineage and differentiation stage-restricted expression, CD77 has been mooted as a therapeutic target in Burkitt lymphoma (BL). The recognition that the globotriaosyl moiety of this neutral glycosphingolipid is a receptor for Escherichia coli-derived Verotoxin-1 (Shiga-Like Toxin-1) offers a potential delivery system for the attack. Here we show that CD77-expressing Group I BL cells which are normally susceptible to activation-induced death on binding Verotoxin-1 B chain are protected in the presence of CD40 ligand. Ectopic expression of either bcl-2 or bcl-xL also afforded resistance to the actions of the B chain. In total contrast, neither of the survival genes nor a CD40 signal - even when acting in concert - protected against killing mediated by the holotoxin. These findings indicate that while therapeutic modalities for CD77 expressing B cell tumors (which include follicular lymphoma) based on the use of Verotoxin-1 B chain might be compromised by the activation of endogenous or exogenous survival pathways, those exploiting the holotoxin should be left unscathed. PMID- 11042674 TI - Activation of protein kinase C relays distinct signaling pathways in the same cell type: differentiation and caspase-mediated apoptosis. AB - Activation of PKC with 5 nM 12-O-tetradecanoylphorbol-13-acetate (TPA) for 72 h in human U937 myeloid leukemia cells is associated with induction of adherence, followed by monocytic differentiation and G0/G1 cell cycle arrest. In this study, we demonstrate that in addition to these effects about 25% of U937 cells accumulated in an apoptotic subG1 phase after TPA treatment. The appearance of these apoptotic suspension cells was detectable throughout the time course of the culture and was independent of TPA concentrations between 0.5 and 500 nM. Experiments with cells synchronized by centrifugal elutriation revealed dominant susceptibility of G1-phase cells to TPA-mediated apoptosis. While adherent cells expressed differentiation markers including the integrin CD11c, this effect was less pronounced in the TPA-treated suspension fraction. Moreover, previous work has demonstrated cell cycle arrest in differentiating U937 cells. Accordingly, PKC activation by TPA treatment was associated with a significant expression of the cdk/cyclin inhibitor p21WAF/CIP/sdi-1 in the adherent population and subsequent G0/G1 cell cycle arrest. In contrast, suspension cells failed to induce significant levels of p21WAF/CIP/sdi-1 after TPA stimulation. Immunoblotting experiments demonstrated no difference in the expression of the pro-apoptotic factors Bax, Bad, and Bak in either control U937 and TPA-treated adherent or suspension cells, respectively. However, anti-apoptotic factors including Bcl-2, Bcl-xL, and Mcl-1 were significantly induced in the adherent population whereas no induction was detectable in the suspension cells. In this context, incubation with the caspase-3/caspase-7 specific tetrapeptide inhibitor DEVD prior to TPA treatment prevented an accumulation of cells in subG1, respectively, demonstrating an involvement of these caspases. Taken together, these data suggest that PKC activation can relay distinct signaling pathways such as induction of adherence coupled with monocytic differentiation and growth arrest, or induction of caspase-mediated apoptosis coupled with the failure to adhere and to differentiate. PMID- 11042675 TI - Bcl-2 and Bax mammalian regulators of apoptosis are functional in Drosophila. AB - Studies of apoptosis in C. elegans have allowed the identification of three genes, ced-3, ced-4 and ced-9. Their products constitute the components of an induction pathway of apoptosis conserved in the nematode and mammals. In Drosophila, homologues have been found for CED-3, CED-4 and CED-9. CED-9 belongs to the Bcl-2 family which includes negative (Bcl-2) and positive (Bax) regulators of apoptosis. The recently discovered Bcl-2 family member named Drob-1 acts as a positive regulator of cell death. To address whether a Bcl-2 anti-apoptotic pathway exists in the fly, we studied the effects of expressing the mammalian genes bcl-2 in Drosophila. In embryos, expression of bcl-2 inhibits developmental and X-ray-induced apoptosis. Expressing bcl-2 or the pro-apoptotic mammalian bax in the developing eye and wing alters these structures, bcl-2 increasing the number of cells, while bax reduces the number of cells. In addition, the functional interaction between Bcl-2 and Bax is conserved. These results indicate that factors necessary for the activity of bcl-2 and bax are present in Drosophila. Therefore, a Bcl-2 pathway for inhibition of cell death may exist in the fly. PMID- 11042676 TI - The X-linked inhibitor of apoptosis (XIAP) prevents cell death in axotomized CNS neurons in vivo. AB - The inhibition of neuronal apoptosis in acute traumatic and ischemic injuries as well as in long term neurodegenerative disorders like spinal muscular atrophy and possibly Alzheimer's disease is a fundamental requirement for a therapeutic strategy. In this study we used an established in vivo model system of induction of neuronal apoptosis in the CNS to evaluate the properties of the X-linked inhibitor of apoptosis protein (XIAP) to inhibit secondary cell death after axonal lesions. We used adenoviral vectors to transduce retinal ganglion cells after axotomy of the optic nerve of adult rats. Vector application was performed at the optic nerve stump so that only the lesioned retinal neurons could be transduced. We found XIAP to be as effective as the viral broad spectrum caspase inhibitor protein p35. These findings suggest that axotomized RGCs degenerate through class II caspase activity and furthermore offer the possibility of using mammalian XIAP protein to inhibit neuronal apoptosis as a basis for a regenerative therapy in the CNS. PMID- 11042677 TI - M1 muscarinic receptors block caspase activation by phosphoinositide 3-kinase- and MAPK/ERK-independent pathways. AB - When PC12 cells are deprived of trophic support they undergo apoptosis. We have previously shown that survival of trophic factor-deprived PC12M1 cells can be promoted by activation of the G protein-coupled muscarinic receptors. The mechanism whereby muscarinic receptors inhibit apoptosis is poorly understood. In the present study we investigated this mechanism by examining the effect of muscarinic receptor activation on the serum deprivation-induced activity of key players in apoptosis, the caspases, in PC12M1 cells. The results showed that m1 muscarinic activation inhibits caspase activity induced by serum deprivation. This effect appeared to be caused by the prevention of activation of caspases such as caspase-2 and caspase-3, and not by the inhibition of existing activity. Muscarinic receptor activation also stimulated the mitogen-activated protein kinase/extracellular signaling-regulated kinase (MAPK/ERK) and phosphoinositide (PI) 3-kinase signaling pathways. The PI 3-kinase pathway inhibitors wortmannin and LY294002, as well as the MAPK/ERK pathway PD98059 inhibitor, did not however suppress the inhibitory effect of the muscarinic receptors on caspase activity. The results therefore suggested that the muscarinic survival effect is mediated by a pathway that leads to caspase inhibition by MAPK/ERK- and PI 3-kinase independent signaling cascades. PMID- 11042678 TI - Resveratrol causes arrest in the S-phase prior to Fas-independent apoptosis in CEM-C7H2 acute leukemia cells. AB - Resveratrol (3,5,4'-trihydroxy-trans-stilbene), in the concentration range of 20 microM and above, induced arrest in the S-phase and apoptosis in the T cell derived T-ALL lymphocytic leukemia cell line CEM-C7H2 which is deficient in functional p53 and p16. Expression of transgenic p16/INK4A, which causes arrest in G0/G1, markedly reduced the percentage of apoptotic cells. Antagonist antibodies to Fas or FasL, or constitutive expression of crmA did not diminish the extent of resveratrol-induced apoptosis. Furthermore, a caspase-8-negative, Fas-resistant Jurkat cell line was sensitive to resveratrol-induced apoptosis which could be strongly inhibited in the Jurkat as well as in the CEM cell line by z-VAD-fmk and z-IETD-fmk. The almost complete inhibition by z-IETD-fmk and the lack of inhibition by crmA suggested caspase-6 to be the essential initiator caspase. Western blots revealed the massive conversion of procaspase-6 to its active form, while caspase-3 and caspase-2 were proteolytically activated to a much lesser extent. PMID- 11042679 TI - Neuroblastoma specific effects of DR-nm23 and its mutant forms on differentiation and apoptosis. AB - DR-nm23 belongs to a gene family which includes nm23-H1, originally identified as a candidate metastasis suppressor gene. Nm23 genes are expressed in different tumor types where their levels have been alternatively associated with reduced or increased metastatic potential. Nm23-H1, -H2, DR-nm23 and nm23-H4 all possess NDP kinase activity. Overexpression of DR-nm23 inhibits differentiation and promotes apoptosis in hematopoietic cells. By contrast, it induces morphological and biochemical changes associated with neural differentiation in neuroblastoma cells. In this study, we show that mutations in the catalytic domain and in the serine 61 phosphorylation site, possibly required for protein-protein interactions, impair the ability of DR-nm23 to induce neural differentiation. Moreover, neuroblastoma cells overexpressing wild-type or mutant DR-nm23 are less sensitive to apoptosis triggered by serum withdrawal. By subcellular fractionation, wild-type and mutant DR-nm23 localize in the cytoplasm and prevalently in the mitochondrial fraction. In co-immunoprecipitation experiments, wild-type DR-nm23 binds other members of nm23 family, but mutations in the catalytic and in the RGD domains and in serine 61 inhibit the formation of hetero multimers. Thus, the integrity of the NDP kinase activity and the presence of a serine residue in position 61 seem essential for the ability of DR-nm23 to trigger differentiation and to bind other Nm23 proteins, but not for the anti apoptotic effect in neuroblastoma cells. These studies underline the tissue specificity of the biological effects induced by DR-nm23 expression. PMID- 11042680 TI - Death at the Dead Sea. PMID- 11042681 TI - A novel germ line juxtamembrane Met mutation in human gastric cancer. AB - Activating mutations in the Met receptor tyrosine kinase, both germline and somatic, have been identified in human papillary renal cancer. Here we report a novel germline missense Met mutation, P1009S, in a patient with primary gastric cancer. The dosage of the mutant Met DNA was elevated in the tumor when compared to its matched normal DNA. Therefore, as with hereditary renal papillary cancer, the mutant Met allele may also be selectively duplicated in the tumor. Different from previously reported Met mutations, which occur in the tyrosine kinase domain, this missense mutation is located at the juxtamembrane domain, and is not constitutively activated. However, following treatment with HGF/SF, the P1009S mutant Met protein, expressed in NIH3T3 cells, displays increased and persistent tyrosine phosphorylation compared to the wild-type Met. Importantly, these cells also form colonies in soft agar, and are highly tumorigenic in athymic nude mice. A second nucleotide change in this region of Met, T1010I, was found in a breast cancer biopsy and a large cell lung cancer cell line. Although this previously reported 'polymorphism' did not stimulate NIH3T3 cell growth in soft agar, it was more active than the wild-type Met in the athymic nude mice tumorigenesis assay, suggesting that it may have effects on tumorigenesis. Met has been shown to be highly expressed in human gastric carcinoma cell lines, and our results raise the possibility that activating missense Met mutations could contribute to tumorigenesis of gastric cancer. PMID- 11042682 TI - High circulating proviral load with oligoclonal expansion of HTLV-1 bearing T cells in HTLV-1 carriers with strongyloidiasis. AB - Adult T cell leukemia (ATLL) develops in 3 - 5% of HTLV-1 carriers after a long period of latency during which a persistent polyclonal expansion of HTLV-1 infected lymphocytes is observed in all individuals. This incubation period is significantly shortened in HTLV-1 carrier with Strongyloides stercoralis (Ss) infection, suggesting that Ss could be a cofactor of ATLL. As an increased T cell proliferation at the asymptomatic stage of HTLV-1 infection could increase the risk of malignant transformation, the effect of Ss infection on infected T lymphocytes was assessed in vivo in HTLV-1 asymptomatic carriers. After real-time quantitative PCR, the mean circulating HTLV-1 proviral load was more than five times higher in HTLV-1 carriers with strongyloidiasis than in HTLV-1+ individuals without Ss infection (P<0.009). This increased proviral load was found to result from the extensive proliferation of a restricted number of infected clones, i.e. from oligoclonal expansion, as evidenced by the semiquantitative amplification of HTLV-1 flanking sequences. The positive effect of Ss on clonal expansion was reversible under effective treatment of strongyloidiasis in one patient with parasitological cure whereas no significant modification of the HTLV-1 replication pattern was observed in an additional case with strongyloidiasis treatment failure. Therefore, Ss stimulates the oligoclonal proliferation of HTLV 1 infected cells in HTLV-1 asymptomatic carriers in vivo. This is thought to account for the shortened period of latency observed in ATLL patients with strongyloidiasis. Oncogene (2000) 19, 4954 - 4960 PMID- 11042683 TI - The effector loop and prenylation site of R-Ras are involved in the regulation of integrin function. AB - The closely related small GTP-binding proteins H-Ras and R-Ras have opposing effects on the regulation of integrin cell adhesion receptors. To gain insight into the properties of R-Ras with respect to the regulation of integrin function and interactions with downstream effectors we performed an analysis of R-Ras variants containing mutations in the effector binding domain and C-terminal prenylation site. We found that the activation of the downstream effector PI 3 kinase was sensitive to mutations in the effector binding domain, as was the binding to the effectors, Ral-GDS, Raf-1 and the novel effector Nore1. Furthermore, specific mutations in the effector binding loop and C-terminal prenylation motif impaired the ability of R-Ras to regulate integrin function in CHO cells. However, the ability of the R-Ras effector loop mutants to bind, and activate known effectors did not correlate with their ability to regulate integrin function. Thus, the known R-Ras effectors are not critical for regulating integrin activation, at least in CHO cells. Consequently, these studies provide insight into the structural basis of the interactions between R Ras and its candidate effectors and suggest the existence of novel mechanisms through which this GTPase could regulate cell adhesion. PMID- 11042684 TI - Estrogen receptor beta acts as a dominant regulator of estrogen signaling. AB - The physiological effects of estrogens are mediated by two intracellular transcription factors, the estrogen receptors (ERs), that regulate transcription of target genes through binding to specific DNA target sequences. Here we describe alterations in cellular responses to different ER agonists and to the anti-estrogenic compound tamoxifen resulting from co-expression of the two ERs in transient co-transfection experiments. Our results demonstrate that ERbeta can act as a negative or positive dominant regulator of ER activity. This is manifested through reduced transcriptional activity at low concentrations of estradiol (E2); increased antagonistic effects of tamoxifen on E2 stimulated activity; and enhanced agonistic action of the phytoestrogenic compound genistein. Furthermore, using chimeric proteins lacking the N-terminal activation function 1 (AF-1), we show that the differential responses of ERalpha and ERbeta to different agonists and antagonists are primarily dictated by inherent differences in the C-terminal ligand-binding domains of the receptors, whereas the magnitude of transcriptional activity is influenced by ERalpha AF-1, but not ERbeta AF-1. The ERalpha AF-1 activity appears to be modulated upon co-expression of both ERs. The alterations in transcriptional activity resulting from co expression of ERalpha and ERbeta are probably due to the formation of alpha/beta heterodimeric complexes. This study demonstrates that co-localization and subsequent heterodimerization of ERalpha and ERbeta may result in receptor activity distinct from that of ER homodimers. PMID- 11042685 TI - Expression of protein tyrosine phosphatase alpha (RPTPalpha) in human breast cancer correlates with low tumor grade, and inhibits tumor cell growth in vitro and in vivo. AB - Tyrosine phosphorylation is controlled by a balance of tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). Whereas the contribution of PTKs to breast tumorigenesis is the subject of intense scrutiny, the potential role of PTPs is poorly known. RPTPalpha is implicated in the activation of Src family kinases, and regulation of integrin signaling, cell adhesion, and growth factor responsiveness. To explore its potential contribution to human neoplasia, we surveyed RPTPalpha protein levels in primary human breast cancer. We found RPTPalpha levels to vary widely among tumors, with 29% of cases manifesting significant overexpression. High RPTPalpha protein levels correlated significantly with low tumor grade and positive estrogen receptor status. Expression of RPTPalpha in breast carcinoma cells led to growth inhibition, associated with increased accumulation in G0 and G1, and delayed tumor growth and metastasis. To our knowledge, this is the first example of a study correlating expression level of a specific bona fide PTP with neoplastic disease status in humans. PMID- 11042686 TI - Rb and p130 regulate RNA polymerase I transcription: Rb disrupts the interaction between UBF and SL-1. AB - We have previously demonstrated that the protein encoded by the retinoblastoma susceptibility gene (Rb) functions as a regulator of transcription by RNA polymerase I (rDNA transcription) by inhibiting UBF-mediated transcription. In the present study, we have examined the mechanism by which Rb represses UBF dependent rDNA transcription and determined if other Rb-like proteins have similar effects. We demonstrate that authentic or recombinant UBF and Rb interact directly and this requires a functional A/B pocket. DNase footprinting and band shift assays demonstrated that the interaction between Rb and UBF does not inhibit the binding of UBF to DNA. However, the formation of an UBF/Rb complex does block the interaction of UBF with SL-1, as indicated by using the 48 kDa subunit as a marker for SL-1. Additional evidence is presented that another pocket protein, p130 but not p107, can be found in a complex with UBF. Interestingly, the cellular content of p130 inversely correlated with the rate of rDNA transcription in two physiological systems, and overexpression of p130 inhibited rDNA transcription. These results suggest that p130 may regulate rDNA transcription in a similar manner to Rb. PMID- 11042687 TI - NPDC-1, a regulator of neural cell proliferation and differentiation, interacts with E2F-1, reduces its binding to DNA and modulates its transcriptional activity. AB - We have previously identified NPDC-1, a neural factor involved in the control of proliferation and differentiation, and we have shown that the stable introduction of NPDC-1 into transformed cells down-regulates cell proliferation both by increasing the generation time and by suppressing transformed properties. The data presented here indicate that, in vitro, NPDC-1 is able to interact with the transcription factor E2F-1 and some cell cycle proteins, such as D-cyclins and cdk2. In addition, two-hybrid experiments in mammalian cells show that the interaction between NPDC-1 and E2F-1 can also occur in vivo. This interaction reduces the binding of E2F-1 to DNA and its transcriptional activity. Taken together, the data suggest that NPDC-1 could influence cell cycle progression and neural differentiation through its association with E2F-1. PMID- 11042688 TI - Differences in mutant p53 protein stability and functional activity in teniposide sensitive and -resistant human leukemic CEM cells. AB - We examined p53 protein stability and DNA damage-induced p53-dependent responses in a human leukemic CEM cell line and two teniposide-resistant sublines, CEM/VM-1 and CEM/VM-1-5 ( approximately 40 and 400-fold resistant to teniposide, respectively). Although all cell lines contain the same p53 mutations at codons 175 (Arg-->His) and 248 (Arg-->Gln), the constitutive levels of p53 were progressively increased with the resistance of the cells to teniposide. By pulse chase experiments, we found that the half-lives of mutant p53 protein were approximately 12, 17, and >30 h in CEM, CEM/VM-1, and CEM/VM-1-5 cells, respectively. The prolonged half-lives of p53 in these cells is consistent with the fact that the protein harbors the indicated mutations. Of note, however, is the fact that the increased p53 protein half-lives in the two drug-resistant cell lines corresponds to a proportional decrease in MDM2 protein levels but an increase in p53-MDM2 binding interactions. This suggests that MDM2-mediated p53 degradation may be altered in our leukemic cell lines. The DNA damage-induced p53 response is fully functional in the drug-sensitive CEM cells containing a mutant p53, but this pathway is attenuated in the drug-resistant cells. Specifically, while the mutant p53 was phosphorylated at serine-15 in response to ionizing radiation in all these cell lines, mutant p53 induction in response to teniposide or ionizing radiation and induction of the p53-target genes, p21 and GADD45 only occurred in the drug-sensitive CEM cells. As assessed by MTT cytotoxicity assay, CEM cells were also significantly more sensitive to ionizing radiation, compared to the drug-resistant cell lines, and this correlated with p53 induction. Collectively, these results suggest that changes in constitutive mutant p53 protein levels, p53-MDM2 binding interactions, and altered regulation of the DNA damage-inducible p53-dependent pathway may play a role in drug- and radiation responsiveness in these cells. PMID- 11042689 TI - Transcriptional control of SPARC by v-Jun and other members of the AP1 family of transcription factors. AB - Transformation of chick embryo fibroblasts by the v-Jun oncoprotein correlates with a down-regulation of the extracellular matrix protein SPARC and repression of the corresponding mRNA. Alteration in SPARC expression has been repeatedly reported in human cancers of various origin, and is thought to contribute to the remodeling of the extracellular matrix during neoplastic progression. Transcriptional control of SPARC is poorly understood. We show here that (i) v Jun-mediated repression of the endogenous SPARC gene is enhanced by Fra2 but alleviated by ATF2, Fra2 and ATF2 being the two major partners of v-Jun in the transformed cells; (ii) high basal activity as well as repression by v-Jun and modulation by Fra2 and ATF2 is restricted to a small proximal fragment (-124/+16) of the chicken SPARC promoter; (iii) the activity of this minimal promoter is modulated by all the AP1 family members known in chickens (c-Jun and JunD; c-Fos and Fra2; ATF2; c-Maf, MafA, and MafB). Taken together these data demonstrate that, at least in avian primary cells, SPARC expression is under the control of the AP1 transcription factor. Further studies with the minimal (-124/+16) promoter fragment are needed to understand how this control takes place at the molecular level. PMID- 11042690 TI - A novel assay for the measurement of Raf-1 kinase activity. AB - Raf-1 is a serine-threonine protein kinase that functions as a central component of the mitogen-activated protein kinase signal transduction pathway. Raf-1 activity is currently assayed in vitro by either measuring 32P incorporation into MEK, Raf-1's only characterized substrate, or by using the phosphorylated MEK to initiate a coupled assay culminating in the phosphorylation of myelin basic protein by MAP kinase. These assays are plagued by a potential lack of specificity in the case of the former, and the time consuming and error-prone nature of the later indirect assay. In this report, we demonstrate a novel single step assay for Raf-1 kinase activity based on phosphorylation of recombinant MEK 1, detected using an activation-specific MEK antibody that recognizes MEK only when specifically phosphorylated by Raf-1 on Ser 217 and Ser 221. The assay readily detected stem cell factor-mediated Raf-1 activation. MEK phosphorylation by immunoprecipitated Raf-1 plateaued at 10 min following initiation of the kinase reaction and was completely dependent on the inclusion of Raf-1. There was a linear correlation between the degree of MEK phosphorylation and the amount of Raf-1 protein immunoprecipitated. In addition to detecting growth factor-mediated activation, the assay was also able to detect paclitaxel-mediated Raf-1 activation. This assay is rapid, sensitive, and specific and therefore is a marked improvement over currently utilized techniques. PMID- 11042691 TI - Age-dependent spontaneous mutagenesis in Xpc mice defective in nucleotide excision repair. AB - DNA damages caused by cellular metabolites and environmental agents induce mutations, that may predispose to cancer. Nucleotide excision repair (NER) is a major cellular defence mechanism acting on a variety of DNA lesions. Here, we show that spontaneous mutant frequencies at the Hprt gene increased 30-fold in T lymphocytes of 1 year old Xpc-/- mice, possessing only functional transcription coupled repair (TCR). Hprt mutant frequencies in Xpa-/- and Csb-/- mice that both have a defect in this NER subpathway, remained low during ageing. In contrast to current models, the elevated mutation rate in Xpc-/- mice does not lead to an increased tumour incidence or premature ageing. Oncogene (2000) 19, 5034 - 5037 PMID- 11042692 TI - Interferon-beta induces S phase slowing via up-regulated expression of PML in squamous carcinoma cells. AB - Type I Interferon (IFN) and all-trans retinoic acid (RA) inhibit cell proliferation of squamous carcinoma cell lines (SCC). Examinations of growth affected cell populations show that SCC lines ME-180 and SiHa treated with IFN beta undergo a specific slower progression through the S phase that seems to trigger cellular death. In combination treatment RA potentiates IFN-beta effect in SCC ME-180 but not in SiHa cell line, partially resistant to RA antiproliferative action. RA added as single agent affects cell proliferation differently by inducing a slight G1 accumulation. The IFN-beta-induced S phase lengthening parallels the increased expression of PML, a nuclear phosphoprotein specifically up-regulated at transcriptional level by IFN, whose overexpression induces cell growth inhibition and tumor suppression. We report that PML up regulation may account for the alteration of cell cycle progression induced by IFN-beta in SCC by infecting cells with PML-PINCO recombinant retrovirus carrying the PML-3 cDNA under the control of the 5' LTR. In fact PML overexpression reproduces the IFN-beta-induced S phase lengthening. These findings provide important insight into the mechanism of tumor suppressing function of PML and could allow PML to be included in the pathways responsible for IFN-induced cell growth suppression. PMID- 11042693 TI - Dual functions of E2F-1 in a transgenic mouse model of liver carcinogenesis. AB - Deregulation of E2F transcriptional control has been implicated in oncogenic transformation. Consistent with this idea, we recently demonstrated that during hepatocarcinogenesis in c-myc/TGFalpha double transgenic mice, there is increased expression of E2F-1 and E2F-2, as well as induction of putative E2F target genes. Therefore, we generated transgenic mice expressing E2F-1 under the control of the albumin enhancer/promoter to test the hypothesis that E2F family members may contribute to liver tumor development. Overexpression of E2F-1 resulted in mild but persistent increases in cell proliferation and death during postnatal liver growth, and no increases in hepatic regenerative growth in response to partial hepatectomy. Nevertheless, from 2 months postnatally E2F-1 transgenic mice exhibited prominent hepatic histological abnormalities including preneoplastic foci adjacent to portal tracts and pericentral large cell dysplasia. From 6 to 8 months onward, there was an abrupt increase in the number of neoplastic nodules ('adenomas') with 100% incidence by 10 months. Some adenomas showed evidence of malignant transformation, and two of six mice killed at 12 months showed trabecular hepatocellular carcinoma. Endogenous c-myc was up-regulated in the early stages of E2F-1 hepatocarcinogenesis, whereas p53 was overexpressed in the tumors, suggesting that both E2F-1-mediated proliferation and apoptosis are operative but at different stages of hepatocarcinogenesis. In conclusion, E2F-1 overexpression in the liver causes dysplasia and tumors and suggests a cooperation between E2F-1 and c-myc oncogenes during liver oncogenesis. PMID- 11042694 TI - The ternary complex factor Net contains two distinct elements that mediate different responses to MAP kinase signalling cascades. AB - The ternary complex factors (TCFs), Elk-1, Sap-1a and Net, are key integrators of the transcriptional response to different signalling pathways. Classically, three MAP kinase pathways, involving ERK, JNK, and p38, transduce various extracellular stimuli to the nucleus. Net is a repressor that is converted into an activator by Ras/ERK signalling. Net is also exported from the nucleus in response to stress stimuli transduced through the JNK pathway, leading to relief from repression. Here we show that ERK and p38 bind to the D box and that binding is required for phosphorylation of the adjacent C-terminally located C-domain. The D box as well as the phosphorylation sites in the C-domain (the DC element) are required for transcription activation by Ras. On the other hand, JNK binds to the J box in the middle of the protein, and binding is required for phosphorylation of the adjacent EXport motif. Both the binding and phosphorylation sites (the JEX element) are important for Net export. In conclusion, specific targeting of Net by MAP kinase pathways involves two different docking sites and phosphorylation of two different domains. These two elements, DC and JEX, mediate two distinct functional responses. PMID- 11042695 TI - Comparative microarray analysis of gene expression during apoptosis-induction by growth factor deprivation or protein kinase C inhibition. AB - The transcriptional response of mouse pro-B cells to two different apoptotic stimuli was investigated. First, interleukin-3 (IL-3) deprivation was used to trigger programmed cell death in IL-3 dependent FL5.12 cells. Alternatively, cells were treated with the protein kinase C (PKC) inhibitor staurosporine. The temporal pattern of gene expression was followed with cDNA microarrays, covering over 8700 different mouse cDNA sequences corresponding to approximately 7900 unique genes. Messenger RNA levels of 315 genes were found to be regulated by more than twofold upon IL-3 removal, while 125 genes reacted to staurosporine treatment. Cross-comparison revealed an intersection of 34 genes similarly regulated in both pathways and thus representing candidates for common apoptosis regulators. For many expressed sequence tags (ESTs) our data suggest for the first time functions in the control of apoptosis, stress response or the cell cycle. IL-3 removal led to the repression of genes required for proliferation and to the induction of genes, linked to apoptotic and signaling pathways. Staurosporine caused predominantly activation of genes, some of which had previously been described to be involved in inflammation. Our findings indicate that cellular responses to both apoptotic stimuli influence various physiological pathways which had not previously been known to be linked. PMID- 11042696 TI - A specific function for phosphatidylinositol 3-kinase alpha (p85alpha-p110alpha) in cell survival and for phosphatidylinositol 3-kinase beta (p85alpha-p110beta) in de novo DNA synthesis of human colon carcinoma cells. AB - We have previously shown an important function of phosphatidylinositol 3-kinase (PI3K)alpha(p85alpha-p110alpha) and PI3Kbeta (p85-alpha-p110beta) for DNA synthesis induced by various mitogens in non transformed fibroblasts and we now report a specific role of these enzymes in human colon cancer cell growth. Using antibodies specific to p110alpha and to p110beta catalytic subunits, increase in PI3Kalpha and PI3Kbeta activities was detected in 15/19 human tumour biopsies relative to adjacent normal mucosa of human colon and bladder. Increase in such activities was also observed in adenocarcinoma cell lines CaCo2, CO115, HCT 116, LS 174T and WiDr relative to non-transformed fibroblasts. Maximal PI3Kalpha activity was observed for LS 174T and PI3Kbeta activity for WiDr cells. This was partly correlated with an increase in p110alpha and p110beta protein levels both in some primary tumours and established cell lines, suggesting that PI3K overexpression is involved in enzymatic deregulation. Functional consequence of such activation was assessed by a microinjection approach. An injection of neutralizing antibody specific to p110beta in WiDr, HCT116 and CO 115 cells inhibited de novo DNA synthesis, whereas antibodies specific to p110gamma had no effect. Neutralizing antibodies specific to p110alpha induced apoptosis, a response that was reverted by treating cells with the caspase inhibitor z-VAD fmk. However anti-p110beta and anti-p110gamma antibodies did not affect cell survival. We concluded that PI3Kalpha and PI3Kbeta play important roles in human colon cancer cell growth with a specific function for PI3Kbeta in de novo DNA synthesis and an involvement of PI3Kalpha in cell survival. PMID- 11042697 TI - The anti-proliferative effects of 1alpha,25(OH)2D3 on breast and prostate cancer cells are associated with induction of BRCA1 gene expression. AB - The anti-proliferative action of the seco-steroid hormone 1alpha, 25 dihydroxyvitamin D3 [1alpha,25(OH)2D3] extends to some, but not all breast and prostate cancer cell lines. By elucidating the molecular mechanisms mediating the sensitivity of these cells, we can identify critical target genes regulated directly or indirectly by 1alpha,25(OH)2D3 and pathways potentially disrupted during transformation. In this study, we demonstrated the induction of expression of BRCA1 mRNA and protein as well as transcriptional activation from the BRCA1 promoter by 1alpha,25(OH)2D3 in the sensitive breast cancer cell line MCF-7. This was not observed in the 1alpha,25(OH)2D3-resistant breast cancer cell line MDA-MB 436. The induction of BRCA1 mRNA was blocked by cyclohexamide. This indicated that transcriptional activation was mediated indirectly by the vitamin D receptor (VDR). Inhibition of VDR protein levels by stable transformation of the anti sense VDR in MCF-7 reduced the sensitivity of MCF-7 to 1alpha,25(OH)2D3 by 50 fold. In addition, the induction of BRCA1 protein and transcriptional activation of a BRCA1 promoter-luciferase reporter construct was abrogated in the stable transformant with the greatest reduction of VDR levels. Examination of other breast and prostate cancer cell lines revealed that sensitivity to the anti proliferative effects of 1alpha, 25(OH)2D3 was strongly associated with an ability to modulate BRCA1 protein. Furthermore, the expression of the estrogen receptor in these cell lines strongly correlated with their sensitivity to 1alpha,25(OH)2D3 and their ability to modulate BRCA1 expression. Taken together, our data support a model whereby the anti-proliferative effects of 1alpha,25(OH)2D3 are mediated, in part, by the induction of BRCA1 gene expression via transcriptional activation by factors induced by the VDR and that this pathway is disrupted during the development of prostate and breast cancers. PMID- 11042698 TI - A serine 37 mutation associated with two missense mutations at highly conserved regions of p53 affect pro-apoptotic genes expression in a T-lymphoblastoid drug resistant cell line. AB - The p53 protein accumulates rapidly through post-transcriptional mechanisms following cellular exposure to DNA damaging agents and is also activated as a transcription factor leading to growth arrest or apoptosis. Phosphorylation of p53 occurs after DNA damage thereby modulating its activity and impeding the interaction of p53 with its negative regulator oncogene Mdm2. The serines 15 and 37 present in the amino terminal region of p53 are phosphorylated by the DNA dependent protein kinase (DNA-PK) in response to DNA damage. In order to verify if specific p53 mutations occur in the multi-drug resistance phenotype, we analysed the p53 gene in two T-lymphoblastoid cell lines, CCRF-CEM and its multi drug-resistant clone CCRF-CEM VLB100, selected for resistance to vinblastine sulfate and cross-resistant to other cytotoxic drugs. Both cell lines showed two heterozygous mutations in the DNA binding domain at codons 175 and 248. The multi drug resistant cell line, CCRF-CEM VLB100, showed an additional mutation that involves the serine 37 whose phosphorylation is important to modulate the protein activity in response to DNA damage. The effects of these mutations on p53 transactivation capacity were evaluated. The activity of p53 on pro-apoptotic genes expression in response to DNA damage induced by (-irradiation, was affected in the vinblastine (VLB) resistant cell line but not in CCRF-CEM sensitive cell line resulting in a much reduced apoptotic cell death of the multi-drug resistant cells. PMID- 11042699 TI - SPI-1 transforming properties depend upon specifically activated forms of the EPOR. AB - Friend erythroleukemia induced in mice by the spleen focus forming virus (SFFV) is a multi-step process. The pre-leukemic phase of the disease results from the abnormal activation of the Erythropoietin (Epo) receptor by the gp55 env gene product of SFFV. Later in disease progression, the emergence of leukemic clones is associated with recurrent genetic events, in particular the activation of the expression of SPI-1, an ETS family transcriptional regulator. We show here that the expression of either SPI-1 or GP55 with the mouse EPOR in avian primary erythroblasts only marginally affects their normal Epo-induced terminal differentiation. In contrast, the co-expression of GP55 and SPI-1 resulted in inhibition of Epo-induced differentiation of EPOR-expressing erythroblasts, promoting instead their proliferation. Co-expression of SPI-1 and GP55 also inhibited the apoptotic cell death program normally induced in response to Epo withdrawal. This cooperation between SPI-1 and GP55 to induce primary erythroblast transformation suggests that progression of Friend erythroleukemia critically depends upon inter-dependent interactions between the molecular events specific of the early and late phase of the disease. PMID- 11042700 TI - Aberrant p27Kip1 promoter methylation in malignant melanoma. AB - p27Kip1 is a regulator of the mammalian cell cycle and a putative tumor suppressor. Distinct altered patterns of p27Kip1 protein expression are found in a variety of human carcinomas, and p27Kip1 expression levels usually correlate directly with disease-free survival. The mechanism(s) by which p27Kip1 expression is reduced or lost during tumorigenesis remains unclear. Specific alterations of the p27Kip1 gene, including mutations and homozygous deletions, are exceedingly rare in human cancers. We have used methylation-specific PCR and bisulfite genomic sequencing to examine the methylation status of p27Kip1 in 61 primary and metastatic tumors and 35 cell lines from patients with malignant melanoma. Dense methylation of a CpG island in the promoter region of p27Kip1 was detected in four of 45 metastatic tumors (9%) and three of the cell lines (9%), including two cell lines established from two different metastases from the same patient. Examination of a naturally occurring, allele-specific sequence variant demonstrated that p27Kip1 methylation is associated with transcriptional silencing in situ. Cell lines with p27Kip1 methylation showed retention of the wild-type allele and detectable p27Kip1 protein whose abundance was reduced compared with normal melanocytes. Collectively, our data suggest that DNA methylation may be a cause of monoallelic p27Kip1 silencing in malignant melanoma, which would support a role for p27Kip1 haploinsufficiency in human cancer. PMID- 11042701 TI - Phosphorylation of the retinoblastoma-related protein p130 in growth-arrested cells. AB - The retinoblastoma family of proteins including pRB, p107 and p130 undergoes cell cycle dependent phosphorylation during the mid-G1 to S phase transition. This phosphorylation is dependent upon the activity of cyclin D/cdk4. In contrast to pRB and p107, p130 is phosphorylated during G0 and the early G1 phase of the cell cycle. We observed that p130 is specifically phosphorylated on serine and threonine residues in T98G cells arrested in G0 by serum deprivation or density arrest. Identification of the phospho-serine and phospho-threonine residues revealed that most were clustered within a short co-linear region unique to p130, defined as the Loop. Deletion of the Loop region resulted in a change in the phosphorylation status of p130 under growth arrest conditions. Notably, deletion of the Loop did not affect the ability of p130 to bind to E2F-4 or SV40 Large T antigen, to induce growth arrest in Saos-2 cells, and to become hyperphosphorylated during the proliferative phase of the cell cycle. p130 undergoes specific G0 phosphorylation in a manner that distinguishes it from pRB and p107. PMID- 11042702 TI - Education of physician on end of life care: Indian perspective. PMID- 11042703 TI - Growth in the first year in children following IAP Policy on Infant Feeding. AB - OBJECTIVE: To study the growth pattern in the first year in children fed according to recommendations of IAP Policy on Infant Feeding. DESIGN: Longitudinal. SETTING: Department of Pediatrics, S.C.B. Medical College Hospital, Cuttack, Orissa. SUBJECTS AND METHODS: 114 infants (68 boys and 46 girls) with birth weight greater than or equal to 2500g from upper and middle S-E status were regularly followed up from birth to 12 mo of age and fed according to recommendations of IAP Policy on Infant Feeding. Mean and standard deviations of weight for age (W/A) and length for age (L/A) and mean Z scores for W/A, L/A and W/L (weight for length) were calculated separately for boys and girls with reference to NCHS-WHO and BFDS data. OBSERVATIONS: Mean Z scores for W/A with reference to NCHS-WHO data showed a positive trend from birth upto the age of 3 to 4 months, subsequently declining upto one year. The Z scores for L/A showed only a minimal downward trend. The W/L Z score remained above the baseline value up to 3 months in boys and 7 months in girls. When BFDS was taken as the reference, W/A Z scores showed consistent positive increments, from birth in girls and 1 mo in boys. L/A Z scores increased from 3 months in boys and 11 months in girls. Using NCHS data as the reference, the percentage of infants below -2SD for weight was 0 to 7% during first 6 months and 14% at 12 months. Ten% were below -2SD for length at 12 months. With BFDS as the reference, the percentage of infants below -2SD for weight was 25% at birth, 5% at 6 months and 12% at 1 yr. For length, it was 12% at birth and 8% at 1 year. The increments in weight and length closely followed BFDS upto 12 mo age. CONCLUSION: The IAP Policy on Infant Feeding results in adequate growth of non low birth weight infants in the first year of life. PMID- 11042704 TI - Changes in nutritional status and morbidity over time among pre-school children from slums in Pune, India. AB - OBJECTIVE: To investigate changes in nutritional status and morbidity over time among pre-school slum children. DESIGN: Longitudinal. METHODS: Children in the age group of 0-5 years from three slums in Pune (n = 845) were studied for a period of two years. Measurement of weight (up to 20 g) and height (up to 0.1 cm), morbidity (in last 7 days) and clinical assessment was undertaken once every four months. RESULTS: Peak prevalence of malnutrition was observed around 18 months and shorter period (3.5 months) of exclusive breastfeeding was probably responsible. Morbidity was generally higher in rainy season and was associated with wasting but not stunting. Gastrointestinal illness and fever contributed 50% of total morbidity days. Higher morbidity affected significantly growth velocities in weight throughout pre-school age. Height velocities were significantly low upto three years of age but there appeared no scope for catch up growth as velocities remained similar thereafter. Higher morbidity in younger children (less than 2 years) led to deterioration of nutritional status over time in 30% to 50% children. CONCLUSION: Shorter period of exclusive breastfeeding results in undernutrition at an early age among slum children. Morbidity further deteriorates the nutritional status PMID- 11042705 TI - Factors associated with severe asthma. AB - OBJECTIVE: To study the role of various factors associated with development and severity of bronchial asthma in children between 5-15 years of age. SETTING: Tertiary Care Medical College Hospital. METHODS: A case control study was carried out during May 96 to April 98. Sixty children each suffering from mild (chronic) and severe asthma (chronic) and 60 controls were enrolled to study the association of various risk factors with development of asthma and for severe disease. RESULTS: On univariate analysis factors associated with significant risk for development of asthma included family history of asthma (p = 0.003), lack of exclusive breastfeeding (p = 0.05), past history of bronchiolitis (p = 0.02), associated allergic rhinitis (p = 0.04) and atopic dermatitis (p = 0.01). For development of severe asthma, associated factors were early onset of symptoms (p = 0.01), family history of asthma in grandparents (p = 0.04) and more than 10 cigarettes per day smoked by any family member. No significant effect of air pollution, overcrowding, pets and passive smoking were found on either development of asthma or it's severity. On multivariate analysis only age of onset below 48 months was associated with severe asthma (OR 2.13, 95% CI 1.00 4.54). Exclusive breastfeeding for more than 4 months was the most protective factor for development of asthma (OR 0.25, 95% CI 0.08-0.70). A strong association between development of asthma and past history of bronchiolitis or tuberculosis (OR 5.26, 95% CI 1.7-16.20) and presence of associated atopic dermatitis or rhinitis (OR 7.5, 95% CI 1.64-34.48) was observed. CONCLUSION: History of associated allergic diseases and past history of bronchiolitis were significantly associated with development of asthma. Exclusive breastfeeding for first 4 months of life was protective. The most significant factor associated with severe asthma was onset of illness before 48 months of age. There was no significant effect of air pollution, over crowding, pets at home or passive smoking on severity of asthma PMID- 11042706 TI - Japanese encephalitis vaccine. PMID- 11042708 TI - Developing health services for children. PMID- 11042707 TI - Recurrent/persistent pneumonia. PMID- 11042709 TI - Role of cetirizine in treatment of eosinophilia. PMID- 11042710 TI - Renal insult in asphyxia neonatorum. PMID- 11042711 TI - Evaluating Bacillus-Calmette-Guerin vaccination by tuberculin skin test response. PMID- 11042712 TI - Endoscopic and histologic evaluation of reflux esophagitis. PMID- 11042713 TI - Breastfeeding in adopted babies. PMID- 11042714 TI - Neuromyelitis optica with transient autonomic disturbances. PMID- 11042715 TI - Myotonia congenita: response to carbamazepine. PMID- 11042716 TI - Juvenile recurrent parotitis. PMID- 11042717 TI - CHARGE association--need for choanostomy. PMID- 11042718 TI - Neonatal pharyngeal pseudo-diverticulum. PMID- 11042719 TI - Cystic hygroma. PMID- 11042720 TI - Trisomy 13 (Patau syndrome). PMID- 11042721 TI - Misguiding guidelines from IAP? PMID- 11042723 TI - Use of BCG vaccine. PMID- 11042725 TI - Acute nitrobenzene poisoning. PMID- 11042726 TI - Tuberculosis control without protection from BCG. PMID- 11042728 TI - Scorpion sting. PMID- 11042730 TI - Can single dose intramuscular dexamethasone replace five day oral prednisolone therapy in mild to moderate asthma cases? PMID- 11042731 TI - A piece of my mind: My name is Jack. PMID- 11042732 TI - JAMA 100 years ago: HOSPITAL ORGANIZATION PMID- 11042733 TI - Access to health care for the rural elderly. PMID- 11042734 TI - Children with mental problems not getting the care they need. PMID- 11042735 TI - Neonatal screening varies by state of birth. PMID- 11042737 TI - Quick uptakes: access to trauma care PMID- 11042736 TI - 2000 Gairdner Foundation International Awards. PMID- 11042738 TI - Quick uptakes: virus heading south PMID- 11042739 TI - Quick uptakes: bipolar brain chemistry PMID- 11042740 TI - Quick uptakes: antibiotics in the ED PMID- 11042741 TI - Inequalities in racial access to health care. PMID- 11042742 TI - Inequalities in racial access to health care PMID- 11042743 TI - Do increased 5-year survival rates in prostate cancer indicate better outcomes? PMID- 11042744 TI - Do increased 5-year survival rates in prostate cancer indicate better outcomes? PMID- 11042745 TI - Do increased 5-year survival rates in prostate cancer indicate better outcomes? PMID- 11042746 TI - Viral load in treatment with antiretroviral therapy and interleukin 2. PMID- 11042747 TI - Viral load in treatment with antiretroviral therapy and interleukin 2 PMID- 11042748 TI - Effects and ethics of sanctions on childhood immunization rates. PMID- 11042750 TI - Effects and ethics of sanctions on childhood immunization rates PMID- 11042749 TI - Effects and ethics of sanctions on childhood immunization rates. PMID- 11042751 TI - Effects and ethics of sanctions on childhood immunization rates PMID- 11042752 TI - Low rate of seropositivity to poliovirus among teenagers in Myanmar: A potential pocket for polio. PMID- 11042753 TI - Measles-mumps-rubella vaccine in the Italian armed forces. PMID- 11042754 TI - Unmet health needs of uninsured adults in the United States. AB - CONTEXT: In 1998, 33 million US adults aged 18 to 64 years lacked health insurance. Determining the unmet health needs of this population may aid efforts to improve access to care. OBJECTIVE: To compare nationally representative estimates of the unmet health needs of uninsured and insured adults, particularly among persons with major health risks. DESIGN AND SETTING: Random household telephone survey conducted in all 50 states and the District of Columbia through the Behavioral Risk Factor Surveillance System. PARTICIPANTS: A total of 105,764 adults aged 18 to 64 years in 1997 and 117,364 in 1998, classified as long-term (>/=1 year) uninsured (9.7%), short-term (<1 year) uninsured (4.3%), or insured (86.0%). MAIN OUTCOME MEASURES: Adjusted proportions of participants who could not see a physician when needed due to cost in the past year, had not had a routine checkup within 2 years, and had not received clinically indicated preventive services, compared by insurance status. RESULTS: Long-term- and short term-uninsured adults were more likely than insured adults to report that they could not see a physician when needed due to cost (26.8%, 21.7%, and 8.2%, respectively), especially among those in poor health (69.1%, 51.9%, and 21.8%) or fair health (48.8%, 42.4%, and 15.7%) (P<.001). Long-term-uninsured adults in general were much more likely than short-term-uninsured and insured adults not to have had a routine checkup in the last 2 years (42.8%, 22.3%, and 17.8%, respectively) and among smokers, obese individuals, binge drinkers, and people with hypertension, elevated cholesterol, diabetes, or human immunodeficiency virus risk factors (P<.001). Deficits in cancer screening, cardiovascular risk reduction, and diabetes care were most pronounced among long-term-uninsured adults. CONCLUSIONS: In our study, long-term-uninsured adults reported much greater unmet health needs than insured adults. Providing insurance to improve access to care for long-term-uninsured adults, particularly those with major health risks, could have substantial clinical benefits. JAMA. 2000;284:2061-2069 PMID- 11042755 TI - Comprehensive follow-up care and life-threatening illnesses among high-risk infants: A randomized controlled trial. AB - CONTEXT: Inner-city high-risk infants often receive limited and fragmented care, a problem that may increase serious illness. OBJECTIVE: To assess whether access to comprehensive care in a follow-up clinic is cost-effective in reducing life threatening illnesses among high-risk, inner-city infants. DESIGN: Randomized controlled trial. SETTING AND PARTICIPANTS: A total of 887 very-low-birth-weight infants born in a Texas county hospital between January 1988 and March 1996 and followed up in a children's hospital clinic. One hundred four infants who became ineligible or died after randomization but before nursery discharge were excluded from the analysis. INTERVENTIONS: Infants were randomly assigned to receive routine follow-up care (well-baby care and care for chronic illnesses; n = 441) or comprehensive care (which included the components of routine care plus care for acute illnesses, with 24-hour access to a primary caregiver; n = 446). MAIN OUTCOME MEASURES: Life-threatening illnesses (ie, causing death or hospital admission for pediatric intensive care) occurring between nursery discharge and age 1 year, assessed by blinded evaluators from inpatient charts and state Medicaid and vital statistics records; and hospital costs (estimated from department-specific cost-to-charge ratios). RESULTS: Comprehensive care resulted in a mean of 3.1 more clinic visits and 6.7 more telephone conversations with clinic staff (P<.001 for both). One-year outcomes were unknown for fewer comprehensive-care infants than routine-care infants (9 vs 28; P =.001). Identified deaths were similar (11 in comprehensive care vs 13 in routine care; P =.68). The comprehensive-care group had 48% fewer life-threatening illnesses (33 vs 63; P<.001), 57% fewer intensive care admissions (23 vs 53; P =.003), and 42% fewer intensive care days (254 vs 440; P =.003). Comprehensive care did not increase the mean estimated cost per infant for all care ($6265 with comprehensive care and $9913 with routine care). CONCLUSION: Comprehensive follow up care by experienced caregivers can be highly effective in reducing life threatening illness without increasing costs among high-risk inner-city infants. JAMA. 2000;284:2070-2076. PMID- 11042756 TI - Primary care safety-net delivery sites in the United States: A comparison of community health centers, hospital outpatient departments, and physicians' offices. AB - CONTEXT: The US primary care safety net is composed of a loose network of community health centers, hospital outpatient departments, and physicians' offices. National data on how the mix of patients and services differ across sites are needed. OBJECTIVE: To develop and contrast national profiles of patient and service mix for primary care. DESIGN, SETTING, AND PATIENTS: Comparative analyses of 3 national surveys of primary care visits occurring in 1994: for data on physician's office visits, the National Ambulatory Medical Care Survey (NAMCS); for hospital outpatient department data, the National Hospital Ambulatory Medical Care Survey (NHAMCS); and for data on community health centers, the Bureau of Primary Health Care's 1994 Survey of Visits to Community Health Centers. A time trend analysis also was conducted using the 1998 NAMCS and NHAMCS. MAIN OUTCOME MEASURES: National estimates of primary care visit rates, types of patient presentation, patient case-mix, disposition of patients, and management interventions in 1994, and compared with 1998 data. RESULTS: The US population made 1.3 primary care visits per person in 1994, which accounted for 43.5% of all ambulatory visits to physicians' offices, community health centers, and hospital outpatient departments. Primary care visits per person were 20% lower for Hispanics and 33% lower for black, non-Hispanic persons compared with white, non-Hispanic persons. Visits to community health centers were more likely to be made by ethnic minorities, patients with Medicaid or no insurance, and rural dwellers than visits made to the other delivery sites. Visits at hospital outpatient departments were made by sicker populations and were characterized by less continuity than the other delivery sites. Controlling for patient mix, visits made to hospital outpatient departments were more commonly associated with imaging studies, minor surgery, and specialty referrals than those made to physicians' offices. In 1998, the US population made an estimated 3. 4 visits per person, 45.6% of which were primary care visits. National estimates of primary care visit rates and patient mix and practice pattern comparisons between hospital outpatient departments and physicians' offices were similar in 1998 and 1994. CONCLUSIONS: Expanding community health centers will likely improve access to primary care for vulnerable US populations. However, enhancing access to of physicians' offices is also needed to bolster the safety net. The greater service intensity and poorer continuity for primary care visits in hospital outpatient departments that we observed raises concern about the suitability of these clinics as primary care delivery sites. JAMA. 2000;284:2077-2083. PMID- 11042757 TI - State scholarship, loan forgiveness, and related programs: the unheralded safety net. AB - CONTEXT: In the mid-1980s, states expanded their initiatives of scholarships, loan repayment programs, and similar incentives to recruit primary care practitioners into underserved areas. With no national coordination or mandate to publicize these efforts, little is known about these state programs and their recent growth. OBJECTIVES: To identify and describe state programs that provide financial support to physicians and midlevel practitioners in exchange for a period of service in underserved areas, and to begin to assess the magnitude of the contributions of these programs to the US health care safety net. DESIGN: Cross-sectional, descriptive study of data collected by telephone, mail questionnaires, and through other available documents, (eg, program brochures, Web sites). SETTING AND PARTICIPANTS: All state programs operating in 1996 that provided financial support in exchange for service in defined underserved areas to student, resident, and practicing physicians; nurse practitioners; physician assistants; and nurse midwives. We excluded local community initiatives and programs that received federal support, including that from the National Health Service Corps. MAIN OUTCOME MEASURES: Number and types of state support-for service programs in 1996; trends in program types and numbers since 1990; distribution of programs across states; numbers of participating physicians and other practitioners in 1996; numbers in state programs relative to federal programs; and basic features of state programs. RESULTS: In 1996, there were 82 eligible programs operating in 41 states, including 29 loan repayment programs, 29 scholarship programs, 11 loan programs, 8 direct financial incentive programs, and 5 resident support programs. Programs more than doubled in number between 1990 (n = 39) and 1996 (n = 82). In 1996, an estimated 1306 physicians and 370 midlevel practitioners were serving obligations to these state programs, a number comparable with those in federal programs. Common features of state programs were a mission to influence the distribution of the health care workforce within their states' borders, an emphasis on primary care, and reliance on annual state appropriations and other public funding mechanisms. CONCLUSIONS: In 1996, states fielded an obligated primary care workforce comparable in size to the better known federal programs. These state programs constitute a major portion of the US health care safety net, and their activities should be monitored, coordinated, and evaluated. State programs should not be omitted from listings of safety-net initiatives or overlooked in future plans to further improve health care access. JAMA. 2000;284:2084-2092. PMID- 11042759 TI - Transforming insurance coverage into quality health care: voltage drops from potential to delivered quality. AB - Although the US health care system is often touted as one of the best in the world, disparities exist in quality of care received by different populations, in different regions, and across different institutions and clinicians. Initiatives to provide access to health insurance have been a major policy tool to ensure that Americans receive high-quality health care. However, availability of insurance coverage does not automatically lead to high-quality care. This article explores points of vulnerability in the US health care system at which the potential to achieve high-quality care can be lost: (1) access to insurance coverage; (2) enrollment in available insurance plans; (3) access to covered services, clinicians, and health care institutions; (4) choice of plans, clinicians, and health care institutions; (5) access to a consistent source of primary care; (6) access to referral services; and (7) delivery of high-quality health care services. Ensuring high-quality health care requires that each of these "voltage drops" be recognized and addressed. JAMA. 2000;284:2100-2107. PMID- 11042758 TI - Access to substance abuse treatment services under the Oregon Health Plan. AB - CONTEXT: The shift to Medicaid managed care has raised numerous concerns about access to publicly funded substance abuse treatment. The implementation of a capitated chemical dependency benefit within the Oregon Health Plan in 1995 provided an opportunity to study the impact of funding mechanisms on access. OBJECTIVES: To determine to what extent access to publicly funded substance abuse treatment changed following the shift to managed care in Oregon and to examine factors associated with that change. DESIGN: Analysis of statewide treatment and Medicaid eligibility data. SETTING AND PATIENTS: All Medicaid-eligible persons aged 12 to 64 years who were enrolled in the Oregon Health Plan during 1994 (88,320), 1996 (170,387), 1997 (160,929), or 1998 (149,877). MAIN OUTCOME MEASURES: Access rates (the number of unique individuals admitted to treatment during a calendar year divided by the average number of enrolled members) computed before (1994) and after (1997) implementation of the capitated benefit. Analyses were replicated with data for 1996 and 1998. RESULTS: The percentage of Medicaid-eligible persons admitted to substance abuse treatment programs during a calendar year increased from 5.5% of the average number of enrolled members per month in 1994 to 7.7% in 1997, following the shift to managed care. For 1996 and 1998, the rates were 6.9% and 7.7%, respectively. Access rates varied considerably among the 7 largest prepaid health plans after adjusting for case mix. Operating characteristics of these prepaid health plans, such as the method of reimbursing treatment providers, were significant predictors of access after controlling for member characteristics. CONCLUSION: According to our analyses, Medicaid-eligible persons in Oregon observed an increase in access to substance abuse treatment after a shift to managed care. JAMA. 2000;284:2093-2099. PMID- 11042760 TI - Ensuring access to health care: the Bush plan. PMID- 11042761 TI - Ensuring access to health care: the Gore plan. PMID- 11042762 TI - Strengthening the US health care safety net. PMID- 11042763 TI - Insuring the uninsured: time to end the aura of invisibility. PMID- 11042766 TI - Ernest Jones (1879-1958). PMID- 11042771 TI - An emigre physician: Dr David (Didier) Roth, homoeopath, art collector, and inventor of calculating machines. PMID- 11042775 TI - Sir James Wylie (1768-1854). PMID- 11042776 TI - Beauperthuy and yellow fever. PMID- 11042777 TI - Nurses must not let themselves be used. PMID- 11042778 TI - Infection control must be the priority of all nurses. PMID- 11042779 TI - Case 22: The incompetent practitioner. Serious concerns about a nurse's basic competencies. PMID- 11042780 TI - Nurse assessment of oral health: a review of practice and education. AB - The assessment of oral health status and related care of patients is a largely neglected area of nursing practice. With the notable exceptions of high-risk patient groups, such as those receiving chemotherapy in neonatal and intensive care units, and in terminal care, few patients enjoy regular, formal, oral assessments and care. Such interventions--nurse administered oral hygiene--should not be reserved only for high-risk groups but ideally be provided to all patients, whether in hospital or in the community, as they can reveal signs and symptoms of oral disease, manifestations of systemic disease, drug side-effects, or trauma; they may also provide important diagnostic clues. This article sets out to emphasize the need for nurse education in oral health care and provides a literature review and introduction to common oral health problems. It also sets out to establish the rationale for assessment in all contexts of patient care. PMID- 11042782 TI - Gerontological nursing. 5: Realizing the future potential. AB - In this, the final article in the series, the authors consider the prospect for the future of gerontological nursing with a special focus on philosophy of care, expert gerontological nurses and evidence-based practice. The article highlights the need for more research and education in gerontological nursing if quality of care is to improve and gerontological nursing is to consolidate its place in the future of nursing practice. PMID- 11042781 TI - Implementing evidence-based practice in infection control. AB - Evidence-based practice is seen as a way of providing more effective health care and is considered to be vital in the current healthcare climate. However, in many areas of practice, and specifically in infection control, there is often little or no evidence to back or refute certain practices. This article looks at ritualistic practices, interventions with indirect evidence to support them and practices with overwhelming evidence in their favour which are not always followed. It is concluded that nurses need to integrate the best available evidence with clinical judgment and ensure that available evidence is disseminated appropriately. PMID- 11042783 TI - Collaboration between nurses and doctors in clinical practice. AB - Several authors have identified collaboration between nurses and doctors as problematic. Benner (1984) stressed that teamwork and collaboration between the disciplines was crucial for both patient care and team morale. The purpose of this article is to evaluate critically and discuss the research studies which have been conducted into the dynamics of the nurse/doctor relationship. A number of characteristics are significant in influencing the collaborative process. These include excellent communication skills, respecting the value of colleagues' roles, the ability to share points of view and trust. PMID- 11042784 TI - Enhancing the nursing role: why nurses want to cannulate. AB - This small qualitative study explored nurses' reasons for undertaking a traditional medical task, that of peripheral intravenous cannulation, and their perceptions as to how this task contributes to their nursing skills. The findings suggest that it benefits the continuity and quality of patient care and provides nurses with personal and professional satisfaction, promoting the delivery of holistic care and the opportunity to initiate autonomous clinical interventions. It is argued that the development of such nursing skills may require professional and organizational support in the form of practice guidelines, protocols and vicarious liability boundaries to enable nurses to practise effectively and efficiently. PMID- 11042785 TI - Diabetes specialist nursing: looking to the future, learning from the past. AB - Diabetes specialist nursing, like many other aspects of nursing, is facing a challenging time in terms of limited resources, staff stress and increasing client caseloads (Naughton and Nolan, 1998). This trend seems set to continue and therefore reflection on and analysis of the role of the diabetes specialist nurse (DSN) is essential so that DSNs can understand their role and meet such challenges proactively. This learning experience (Jarvis, 1992) will also enable DSNs to communicate effectively their role within their teams and to their peers and managers. The author contends that many of the problems currently faced by DSNs can be identified in the initial reactive creation and development of the DSN role which, in some cases, was 20 years ago. Therefore, this article discusses how the past could be influencing the present and future for diabetes specialist nursing. It cannot attempt to provide an exhaustive discussion, but will focus on the following major aspects: DSN titles and commencement; role entry and subroles; and role set and role definition. The article concludes with a vision for the future, informed by this analysis of the past. PMID- 11042786 TI - An evaluation of the impact of lecturer practitioners on learning. AB - The aim of this study was to determine if nursing diploma students recognized a difference in the nature of classroom teaching between lecturer practitioners and university-based senior lecturers in nursing. The study generated both qualitative and quantitative data from a sample of 117 student nurses completing an elderly care module. Analysis of the data suggested that the students associate lecturer practitioners with a realistic, evidence-based presentation of contemporary nursing practice, which they found both academically liberating, and efficacious to their practical training. Such associations were less apparent when analysing student responses to senior lecturer teaching. The study underlines the potential of lecturer practitioners to make a positive and far reaching contribution to the future of nurse education. PMID- 11042787 TI - Confidentiality. 14: Powers of the police and access to information. AB - Staff nurse Paula Rose was on duty in the accident and emergency (A&E) department early one Sunday morning when a police constable came in asking for details of any patient who had been brought in and for whom there was evidence of a fall from height. The constable explained that a girl had been assaulted and raped while in her bedroom by an assailant who came in through the window. Her screams brought help, but he made his escape from the window and she believed that he had fallen and may have suffered fractures or bruises. PMID- 11042788 TI - Wound care: Askina Transorbent and Askina Biofilm Transparent. AB - Selecting the correct dressing for a wound can be complicated as there are so many products from which to choose. The selection process is made easier once the healthcare professional has performed a holistic assessment. This article discusses some of the principles of wound management and the efficacy of Askina Transorbent and Askina Biofilm, two new products from B/Braun. Both of these maintain the moist environment required for optimum wound healing to take place. PMID- 11042789 TI - A nursing career in the RAF. PMID- 11042790 TI - Web watch. PMID- 11042791 TI - New nurse competencies: are they adequate? PMID- 11042792 TI - Who cares for the carers? PMID- 11042793 TI - Golden slumbers. AB - It is interesting to listen to theatre staff who have experienced surgery themselves. Sometimes they have totally changed their practice as a result of their experience, or at any rate have adjusted it. Jeannette has chosen on this occasion to share her distinctive patient's eye view of her trip behind the doors and to pass on her views on anaesthetics. Of course, Jeannette does it all in her own inimitable style, and other patients will not formulate their thoughts in quite the same fashion, nor deliver the message so amusingly ... but how often do we ask the patients for their views? Read the message and sing out.... PMID- 11042794 TI - Being old is detrimental to your health. PMID- 11042795 TI - Postoperative pain management in paediatrics. AB - Pain is defined by the International Association for the Study of Pain as 'an unpleasant sensory and emotional experience associated with actual or potential tissue damage or described in terms of such damage'. It is now accepted that pain should be anticipated, and safely and effectively controlled, in all children, whatever their age, maturity or severity of illness (Fisher & Morton 1998). PMID- 11042796 TI - The support worker in theatres. AB - This article will consider various aspects of the use of the support worker in theatre and will propose that the support worker can undertake the scrubbed role. The position of NATN is that it 'does not recommend that the role of the support worker encompasses the scrub role' (NATN 1996): it will be argued here that this stance is based on arguments which are fundamentally flawed. PMID- 11042797 TI - Library resources and getting the best out of them. AB - Nurses are required to undertake professional updating, PREP (UKCC, 1994), and are increasingly under pressure to ensure that they provide evidence-based healthcare (DoH, 1996). Proliferation of the literature, the move from printed to electronic sources of information and shortage of time may all be seen as barriers to nurses who wish to gain access to information to support these activities. Libraries are important gateways to this information and this article is a brief guide to making the most of library resources. PMID- 11042798 TI - Implications of Working Time Regulations for theatre nurses working in the NHS. AB - The European Working Times Regulations have implications for staff throughout the NHS. In particular, theatre staff and others working on call and shifts need to be aware of the implications of these regulations. In this article, Jon Richards, National Officer for Unison, explains the background to and implications of these regulations for theatre staff. PMID- 11042799 TI - Two gloves or not two gloves that is the question. AB - Surgical gloves were first introduced in 1889, when Halstead wanted to protect his scrub nurse from dermatitis caused by contact with mercuric chloride (Dodds et al 1990). The use of gloves then became more widespread, principally to protect surgical teams whilst operating on infected patients (Dodds et al 1990). Later, focus changed to protecting the wound from microbial contamination. PMID- 11042800 TI - Forward waiting in theatre. AB - Waiting to go into theatre on the day of your surgery is probably the most stressful time for patients. What happens to patients during this crucial time can make a tremendous difference to the whole experience of going to theatre. Heidi Belbin and her colleagues realised that there was a problem at Lincoln City Hospital with patients being kept waiting at theatre reception and were concerned that the quality of patient care in this respect was not acceptable. A plan was developed to improve this situation, and the story of what was done is told in this article. The project was entered for the 3M/NATN award, and earned a commendation from the judges. PMID- 11042801 TI - Pain management in recovery. AB - I make no mistake in mixing anaesthetic care with recovery room nursing, as it is my belief that the two are linked. So this month we look at pain in the recovery room--or is this the only place to which this article is relevant? You decide. Pain is something which, quite rightly, occupies the minds of perioperative practitioners greatly. It is only human, and humane, to use our expertise to alleviate, control and understand it. In this piece, Trudi Starritt from St John's Hospital, Livingston, explores the concept of pain and pain relief from the recovery nurse's perspective. PMID- 11042802 TI - Oncology nurses find radiation setting exciting and challenging. PMID- 11042803 TI - [Nodulation as a model for studying differentiation in higher plants]. AB - The stages of the legume-rhizobial symbiosis and nodule structure in various legume plants are briefly reviewed. Modern data on the mechanisms involved in the control of nodule initiation and morphogenesis are considered. PMID- 11042804 TI - [Satellite DNA and disease--possible mechanisms. Minisatellite instability]. AB - The main possible molecular mechanisms of minisatellite DNA instability are reviewed and compared with those of the trinucleotide repeat instability. Evidence indicating that some human diseases are associated with minisatellite DNA instability is presented including data on minisatellite DNA expansion. PMID- 11042805 TI - [Divergence of the cytochrome b gene in grouse species]. AB - A 720-bp cytochrome b gene fragment of the mitochondrial genome was sequenced in ten species and seven potential subspecies of grouse. Of 187 variable sites detected, 140 were parsimony informative. The distribution of nucleotides in three positions of codons had a similar pattern with and did not fundamentally differ from that of the nucleotide composition of this gene in other animals, including birds. The nucleotides found at the first codon position were shown to be uniformly distributed, whereas the third position was characterized by a significant decrease in the amount of guanine and, to a lesser extent, thymine. Based on the data on the nucleotide sequences of the cytochrome b gene fragment, a phylogenetic tree was constructed. This tree fits well the morphological and ecological differentiation of the species studied. PMID- 11042806 TI - [Phenotypic changes in transgenic tobacco plants with an antisense form of the hmg1 gene]. AB - Tobacco plants were genetically transformed with the Arabidopsis thaliana heterologous hmg1 gene encoding 3-hydroxy-3-methylglutaryl-CoA reductase, a key enzyme involved in the metabolism of terpenoid compounds. The hmg1 gene was inserted under the control of the 35S RNA double promoter from the cauliflower mosaic virus (CaMV 35S) both in direct and reverse orientation relative to the promoter. DNA analysis by polymerase chain reaction (PCR) and Southern blotting confirmed the transgenic nature of the tobacco plants obtained. DNA-RNA hybridization revealed expression of the hmg1 gene in these tobacco plants. The plants transformed with the antisense copy of the hmg1 gene differed from the control plants in delayed development and in flower color and shape. PMID- 11042807 TI - [Genetic connections between some species of Mytilidae (Mollusca: Bivalvia) from the northern part of the Pacific Ocean]. AB - In 1978 and 1999, seven and eight species of Mytilidae (Mollusca: Bivalvia) were analyzed using gel electrophoresis. Mean heterozygosity per individual (Hobs and Hexp) and genetic distances (Rogers' DR, Nei's DN, and others) were estimated for 21 and 24 allozyme loci. Mytilus modiolus had the highest variation among the species examined. Genetic distances were lowest for the M. trossulus-M. galloprovincialis species pair: DR = 0.147, DN = 0.078. Overall, five species of the genera Mytilus and Crenomytilus were genetically closer to each other (DR = 0.147, DN = 0.078) than to the remaining three species of this group (DR = 0.807, DN = 2.243). The relationships among the species were examined using cluster analysis and parsimony methods. The densest clusters in the dendrograms consisted of (1) M. trossulus and M. galloprovincialis and (2) M. coruscus, M. californianus, and M. grayanus. These two clusters form a larger cluster (3), which comprises all representatives of the nominal genus Mutilus and C. grayanus. The Mytilus-Crenomytilus cluster is consecutively joined by Adula falcatoides, Mytilus modiolus, and Septifer keenae. According to Nei's genetic distances DN, the time of divergence between M. trossulus and M. galloprovincialis is 0.8-1.6 Myr; between M. californianus and C. grayanus, it is approximately 9 Myr; and between M. coruscus and the latter pair, it is 13 Myr before present. Two representatives of the Mytilus ex gr. edulis complex diverged from the Mytilus Crenomytilus group of large-size Pacific species about 20 Myr ago. These results are in good agreement with paleontological data and indicate a relatively recent origin of the Mytilus ex gr. edulis complex. The results obtained can be used in systematics and phylogeny of modern Mytilidae. PMID- 11042808 TI - [Isolation of DNA from museum exhibits of butterflies (Lepidoptera, Papilionidae) and PCR analysis with random and universal genes-specific primers]. AB - The effect of the duration of storage of entomological material on DNA preservation was estimated. The results of the optimization of conditions for the analysis of random amplified polymorphic DNA in a polymerase chain reaction (RAPD PCR) are presented as applied to the DNA of lepidopterans of the family Papilionidae. RAPD patterns are shown for the first time in Atrophaneura alcinous and four species of the genus Parnassius (sensu lata). The applicability of museum specimens of butterflies for RAPD analysis was demonstrated. The results of PCR analysis using DNA obtained from different collection specimens stored for up to five years were compared. The authenticity of DNA obtained from collection specimens was proved using PCR with universal primers, which are specific to the COI and COII cytochrome genes of mitochondrial DNA (mt DNA). The lengths of individuals gene fragments obtained by the amplification of both museum and live specimens were 800 and 1600 bp. The conservative regions of mitochondrial genome were shown to be slightly different in two A. alcinous subspecies. PMID- 11042809 TI - [Change in gene action, genotype ranks, and results of selection in a Johannesen populations under the effect of environmental factors]. AB - Eight mapped mutants (er, tfl, fb, fca, fd, fe, fg, and fy) of arabidopsis (Arabidopsis thaliana (L.) Heynh.) were chosen as the model for investigation of plant response to vernalization and day length. Six quantitative characters were recorded: the number of days from sowing until flowering, plant weight, stem height, number of rosette leaves, number of cauline leaves, and total leaf number. The genotypes responded differently to environmental changes, which altered the effects of genes on quantitative traits, genotype ranks, and selection consequences. A certain set of environmental factors may result in selection in favor of one genotype, and another set may result in selection for another genotype, with direction and intensity of selection being the same. PMID- 11042810 TI - [Absence of heterosis in hybrid corn. A genetic analysis of the effect]. AB - The genetics of the absence of heterosis in hybrid plants involving lines TS11 and P22 was studied. Segregation of hybrids (TS11 x 092) x P22, (TS11 x A619) x P22, and (TS11 x 19-3-3) x P22 for plant height was analyzed. The distribution pattern was compared with that of hybrid TS11 x P22, in which hybrid vigor was not observed or was minimal. It was shown that, in 1993 and 1996, the absence of heterosis in hybrid plants was controlled by two recessive genes. In 1998, only one gene was identified. It was suggested that lines TS11, P22, and ST156 may be used to identify genes that determine heterosis in maize. PMID- 11042811 TI - [Effect of chronic irradiation on the genetic structure of natural populations of Arabidopsis thaliana (L.) Heynh]. AB - The genetic structure of Arabidopsis thaliana populations from in the Chernobyl zone of radioactive contamination was studied. The dynamics of genotypic and allele frequencies of eight polymorphic loci were described. Analysis of the results obtained and earlier data revealed selection for loci Per1 and Acp2. Sharp fluctuations of allele frequencies were observed in consecutive generations of chronically irradiated A. thaliana populations. These fluctuations led to replacement of the most frequent alleles, including those close to fixation. PMID- 11042812 TI - [Allozyme variability and genetic divergence of Pacific trout (species Parasalmo) from western Kamchatka]. AB - Genetic variation in several populations of Parasalmo (Onchorhyncus) mykiss from Kamchatka was examined on the basis of data obtained by the author and results from literature. Three (sSOD-1*, LDH-C*, and EST-1*) out of 44 protein-coding loci were highly polymorphic. Low-frequency alternative alleles occurred at sAAT 1,2*, LDH-A2*, EST-5*, IDDH-1,2*, and sMDH-1,2*. The results of the present work and comparison with evidence on North American species indicated that, in the Kamchatka part of the range, P. mykiss is represented by several populations carrying unique alleles and forming a genetically independent group. Gene exchange between North American and Western Kamchatka populations is mainly determined by straying in the feeding stock of the American coastal form entering the Sea of Okhotsk. The genetic divergence and mean heterozygosities in the Kamchatka populations were low (D magnitude of 0.0002-0.0275; HS magnitude of 0.011-0.0371). The difference between the Western Kamchatka populations from the North American coastal form was so small (D magnitude of 0.0109-0.0241) that these forms clustered together. Genetic divergence of the Kamchatka populations and the inland North American P. mykiss is an order of magnitude higher (D magnitude of 0.1973-0.2367). PMID- 11042813 TI - [On the phylogenetic relationship of Corvinae birds (Aves, Corvidae) from data of partial sequencing of cytochrome b gene mitochondrial DNA]. AB - To establish phylogenetic relationships within the corvine birds at the interspecific and intergeneric levels, the sequence of the mitochondrial DNA cytochrome b gene was analyzed. The NJ, UPGMA, and MP trees showed similar clustering. Relationships between the jungle crow, on the one hand, and the rook and Australian raven, on the other hand, were closer than between the jungle crow and the hooded and carrion crows. Mitochondrial genome of Australian raven displayed the closest similarity to the ancestral genome of the genus Corvus. Populations inhabiting the eastern part of the carrion crow C. corone orientations area were statistically significantly subdivided into three lineages. These data also confirmed the hypothesis on the location of the carrion crow ancestral lineage in the southeastern part of the area. In general, the transition and transversion substitution levels, their relationships, and distribution over codon positions were similar to that already reported for birds. Synonymous transitions in the third codon position were the prevailing substitution type. Using standard calibration scales, the time of divergence between species and genera within the corvine family was estimated to be 3.1-4 and 3.8-8.8 Myr, respectively. The divergence time between the examined corvine birds and birds of paradise constituted from 8 to 10 Myr. PMID- 11042814 TI - [Relationship between polymorphism C677T of the methylene tetrahydrofolate reductase gene with clinical symptoms of coronary atherosclerosis]. AB - Association of the methylenetetrahydrofolate reductase gene (MTHFR) C677T missense mutation (substitution of cytosine by thymine at position 677) with coronary artery disease (CAD), as well as with blood levels of various lipoprotein fractions, systolic pressure (SP), diastolic arterial blood pressures (DP), and body mass index (BMI) in patients with angiographically verified CAD and in a control group. The affected and control subjects did not differ substantially with respect to genotypic and allelic frequencies. The MTHFR gene polymorphism was not associated with variation in either total cholesterol (TC), very-low-density-lipoprotein cholesterol (VLDLC), low-density-lipoprotein cholesterol (LDLC), high-density-lipoprotein cholesterol (HDLC), or triglyceride (TG) levels. SP and DP in subjects with different MTHFR genotypes did not differ significantly. BMI tended to correlate with the C677T polymorphism of the MTHFR gene (0.05 < P < 0.1). C677T mutation frequencies in western Siberia were determined for the first time; they are about the same as in most European populations. PMID- 11042815 TI - [Correlation of results of immunocytochemical, electrophysiologic, and molecular studies in patients with cystic fibrosis]. AB - In 13 cystic fibrosis (CF) patients of 5 to 23 years of age with a known mutation spectrum of gene CFTR, sweat chloride values and nasal-potential differences (NPD) were measured and localization characteristics of the protein product of gene CFTR in the cells of nasal epithelium were studied. Sweat Chloride values were normal or boundary (24 to 62 mM/l) in six CF patients. In seven CF patients, these values were significantly above the estimates for the control group. On average, the NPD values were -44.7 +/- 2.2 mV (from -32.5 to -68.9 mV) and -17.2 +/- 1.8 mV (from -6.8 to -30.2 mV) in CF patients and the control group, respectively. Histochemical studies clearly revealed the localization of the CFTR protein on the apical membrane of the nasal epithelium. Depending on the type of mutation, the protein product of gene CFTR was either absent or regularly distributed in the cytoplasm in CF patients; it was not detected in the apical membrane. Thus, NPD measurements and the analysis of the localization of the protein product of gene CFTR in scrapes of nasal epithelium were shown to be additional, highly informative methods of CF diagnostics. PMID- 11042816 TI - [Calculation of the morbid risk in genetic-epidemiologic studies of age-dependent diseases]. AB - Distributions of age at onset are widely used in the genetic epidemiology of age dependent diseases. Examples are estimation of recurrent risks in genetic counselling and testing genetic hypotheses in segregation and linkage analyses. In this study, morbidity parameters are defined, including age-specific morbidity rates, morbidity net risk (incidence), and cumulative incidence (population risk, an integrated measure of population susceptibility to the disease at the moment of the study). Age-specific morbidity risks are calculated from the respective morbidity rates, which are analogous to mortality rates used in demography. Population data typically used for calculation of morbidity rates are discussed. Methods of calculation of morbidity rates based on the data of single and interval epidemiological studies are described. Methods for calculating standard errors of these parameters, estimating their statistical reliability, and testing statistical hypotheses are discussed. PMID- 11042818 TI - The early international dental congresses. AB - The importance of the dissemination of scientific and clinical advances brought about by the early international dental congresses cannot be underestimated. The congresses in 1889, 1893, 1900 and 1909 set the pattern for discussion and association for the rest of the twentieth and, now, the beginning of the twenty first century. PMID- 11042817 TI - The FDI's first ten years, 1900-1910. Federation Dentaire Internationale. AB - On the centenary of the foundation of the FDI and the fiftieth anniversary of the International Dental Journal, the editor looks at the beginnings of the Federation. This paper sets out to aid the understanding not only of the early history of the FDI but also the social, scientific and political background against which it was born and nurtured during its tentative, early years. PMID- 11042819 TI - World dental demographics. AB - AIMS: To collect basic data regarding dental workforce and dental education in all countries of the world and to make this data available on the FDI's Website. METHOD: A postal questionnaire survey. SOURCES OF INFORMATION: Member Associations of the FDI and governmental agencies. RESULTS: Responses were received from 73 countries with a reported total number of dentists of 703,947. A comparison between the figures now reported and the figures published in 1990 shows that the total number of dentists in these countries has increased by 27.8 per cent over the ten year period. No correlation was found between the population per dentist figures and the GNP of the countries. In the reporting countries, there were 550 dental schools. A comparison between the figures now reported and the figures published in 1990 shows that the total number of dental schools in these countries has increased by 42.6 per cent over the ten year period. The total number of dental hygienists was reported to be 181,336 and the total number of dental technicians in these countries was reported to be 252,004. PMID- 11042820 TI - Systematic root cause analysis of adverse drug events in a tertiary referral hospital. AB - BACKGROUND: Adverse drug events (ADEs) occur frequently, and serious ADEs are associated with mortality or prolonged morbidity. As many ADEs are preventable, identification and modification of systems and processes that permit ADEs has the potential to reduce the rate of ADEs. METHODS: Root cause analysis was systematically employed in a blame-free fashion to investigate the patterns of serious ADEs that occurred during a 29-month period at Hermann Hospital (Houston), and process improvements were implemented on the basis of these findings. The consistently nonpunitive responses to the results of the initial and subsequent root cause analyses was gradually seen, accepted, and ultimately embraced by the hospital staff. RESULTS: The most commonly identified root causes were environmental factors (for example, increased census, increased acuity, change of shift) and staffing issues (for example, personnel new to a unit). Policy changes that led to increased use of forcing or constraining functions (for example, removal of concentrated intravenous potassium solutions from floor stocks) and better personnel support (for example, early awareness and response to localized increases in census and acuity) were particularly effective. Although limited by our lack of active surveillance and not necessarily directly due to the process changes that we implemented, the rate of voluntarily reported serious ADEs/100,000 patient days decreased during this time from 7.2 to 4.0, a decline of 45% (p < 0.001). CONCLUSION: Systematic application of root cause analysis followed by implementation of process changes that target the underlying cause(s) of each event can be successfully implemented in a large hospital. PMID- 11042821 TI - The strategic use of outcome information. AB - BACKGROUND: Most health care executives see outcome measurement as a technical or tactical matter rather than as a strategic tool. Accordingly, provider investment in outcome measurement and management is relatively small. Nevertheless, outcome information can be key to achieving an organization's strategic objectives. Advances in risk adjustment and improvements in technology for data collection and analysis have made outcome measurement a practical tool for individual hospital use. CASE STUDIES: Strategically integrated outcome measurement efforts can give providers a competitive advantage over organizations that only use outcomes tactically. One of the best examples of an acute care provider that has used outcome information for strategic advantage is Intermountain Health Care (IHC; Salt Lake City). In 1997 IHC made clinical quality and outcomes the primary focus of its five-year strategic plan. To support the new strategy IHC's board of trustees approved the development of an outcome information system that generated data along clinical processes of care and the creation of a new management structure to use these data to hold professionals accountable and to set and achieve clinical improvement goals. From 1996 to 1999, IHC's share of the commercial health care market in Utah increased from roughly 50% to about 62% of the market, with the result that it has stopped actively marketing its services. DISCUSSION: Health care executives will not willingly invest in outcomes until they believe that they have business value. Therefore, making the business case for outcomes can help improve the quality of health care and the lives of individuals. PMID- 11042822 TI - Preventing home health nursing assistant back and shoulder injuries. AB - BACKGROUND: Franklin County Home Health Agency (St Albans, Vermont) undertook a performance improvement project in 1996 to reduce employee injuries. A review of recent injuries led to the prevention of licensed nursing assistants' (LNAs') back and shoulder injuries as the first priority. Root causes of injuries were agency communication, employee training, patient home environment, nursing assistant body mechanics, and failure to use safety measures. Given that injury causality is complex and multifactorial, a variety of improvement strategies were implemented over the following two to three years. IMPLEMENTATION OF POTENTIAL SOLUTIONS: Short-term (a few months), mid-term (six months), and long-term (one year) potential solutions to the LNA back and shoulder injury problem were charted. Safety and health training was the major focus of the team's short-term plan. Risk management forms were to be used to identify and follow up on hazardous situations. RESULTS: Project plans that were successfully implemented included revision of LNA plans of care, standardization of the return-to-work process after injury, development of guidelines for identifying unsafe patient lifts and transfers, improved follow-up of employee reports of injury-risk situations in patient homes, improved body mechanics screening of new employees, and a stronger injury-prevention training program for current employees. A less successful initiative was aimed at collecting more data about injuries and causal factors. Employee injuries were gradually reduced from 4-10 per quarter to 0-3 per quarter. CONCLUSIONS: Injury prevention requires commitment, persistence, and patience--but not expensive improvements. Multiple interventions increase the chances of success when there are many root causes and lack of evidence regarding the effectiveness of various approaches. PMID- 11042823 TI - Quality improvement in a managed care organization from a medical director's perspective: an interview with Bruce Perry. Interview by Douglas Roblin. AB - BACKGROUND: In late 1994 the Quality Forum commissioned the Interdisciplinary Prevention Committee [IPC]. One of the IPC's charges was to identify priorities for QI in preventive health services. The IPC established priorities through a review of scientific literature, identification of national and state health initiative priorities, and consideration of what services to establish as priorities and of the practicality of implementing low-cost interventions to achieve specific QI goals. Breast cancer screening was selected as a top-ten priority for guideline development and for focused intervention because of the disease's prevalence, morbidity, and mortality and because of the fact that it is most treatable and curable when it is found early through routine screening. The national HEDIS (Health Plan Employer Data and Information Set; National Committee on Quality Assurance (NCQA), Washington, DC] result of 71%, reported in May 1995, provided our baseline performance measurement. This result fell short of our goal of being in the 90th percentile of performance on each HEDIS effectiveness of care measure. In August 1995 the Quality Forum accepted the IPC's recommendations, which had been endorsed by the department of medicine. These recommendations emphasized the importance of annual clinical breast exam and mammography for women of targeted age groups. In November 1997, a new "Excellence in Quality: HEDIS Improvement Team" began work. Its charge was to undertake analyses of underlying causes of reduced performance and to develop additional steps to improve performance by changes in care delivery processes in 1998. In March 1998 the Quality Forum's executive committee designated breast cancer screening one of the six organizationwide quality priorities for 1998 and designated two "owners" who would be accountable for this performance--the chief and director of radiology. RESULTS: The screening rate increased from 73.8% in 1996 to 84.0% in 1999. National benchmarks [90th percentile] in 1998 were 81% for commercially insured members and 84% for Medicare members. The 84% screening rate made the Georgia region the Kaiser Permanente national leader and put the region in the top 10% of all health plans in the United States. CONCLUSIONS: The program has achieved these results with a broad array of activities: Saturday hours, mobile mammography, medical record reminders (fuschia-colored inserts), patient and physician reminders, call-center outreach, provider feedback on performance, and provider financial incentives. Several of these innovations demonstrate the ability to integrate improved care management into evolving service delivery in Kaiser Permanente--such as use of call-center technologies and redesign of primary care delivery. While we cannot point to any one of these innovations as a key driver of improvement, it is clear that substantial improvements in care delivery can be achieved. All these activities are relatively low cost and easily implemented in other managed care organizations and in other areas of medical care. PMID- 11042824 TI - An analysis of the effect of age on implantation rates. AB - PURPOSE: To evaluate implantation rate as a function of age. METHODS: A total of 1621 consecutive cycles of IVF were evaluated for implantation as a function of age at The New York Hospital/Cornell Medical Center. RESULTS: An overall implantation rate of 23.3% (1328/5691) was found. The implantation rate as a function of age decreased in a nonlinear fashion. Implantation remained constant until the age of 35 and then decreased in a significantly, linear fashion by 2.77% per year (P < 0.001, R2 = 0.975). A formula to predict implantation rates for a given age was developed: Implantation rate = -119.352 + (9.985 x Age - (0.164 x Age2)). CONCLUSIONS: We have demonstrated that implantation rates remain constant until the age of 35 at which time a linear decrease of 2.77% per year is observed. PMID- 11042826 TI - Effects of electric stimulation on bovine oocyte activation and embryo development in intracytoplasmic sperm injection procedure. AB - PURPOSE: This study was carried out to investigate the efficacy of electric stimulation before and/or after intracytoplasmic sperm injection (ICSI) on bovine oocyte activation and embryo development. METHODS: The oocytes were treated with electric shock before (B), before and after (B&A), and after (A) sperm injection. In each group, sham ICSI (ICSI-s) was performed to exclude the effect of parthenogenesis (B ICSI-s, B&A ICSI-s, and A ICSI-s). An electric pulse was applied with a single direct current (DC) pulse (0.8 kV/cm, 70 microseconds). RESULTS: One pronucleus (PN) formation in the B&A ICSI-s group was slightly higher than that found in B and B&A ICSI group; however, the difference was not significant. Two PN formation in B&A ICSI group was higher than that found in sham ICSI groups (P < 0.05). There were no differences among treatment groups in the cleavage rate; however, morulae and blastocyst formation in the B&A embryos was significantly higher than that of other groups (P < 0.05) and got pregnant. CONCLUSIONS: Electric stimulation before and after injection was an effective method in inducing bovine oocyte activation and in sustaining embryo development to the morulae and blastocyst stage. PMID- 11042825 TI - Increased incidence of meiotic anomalies in oligoasthenozoospermic males preselected for intracytoplasmic sperm injection. AB - PURPOSE: Based on data from the literature, to detect the possible presence of an increased frequency of meiotic anomalies in oligoasthenozoospermic (OA) patients preselected for intracytoplasmic sperm injection. METHODS: Meiotic studies in as many successive patients with a clinical indication for a diagnostic testicular biopsy as needed to complete at least 100 cases with a severe OA (motile sperm concentration < or = 1.5 x 10(6)/ml). RESULTS: An increased incidence of meiotic anomalies was found in 102 patients with a severe OA (17.6%) compared to the mean for 105 patients with other etiologies in the series (5.7%) or the mean for patients reviewed in the literature (6.5%). CONCLUSIONS: Patients with a severe OA have a higher incidence of synaptic anomalies. This may result in the malsegregation of chromosomes at meiosis I, producing abnormal sperm, and could explain the high incidence of sterility and some cases of abortion (in two thirds of the couples with abortions the husband had meiotic anomalies) in this group. PMID- 11042827 TI - Number and size of antral follicles as predictive factors in vitro fertilization and embryo transfer. AB - PURPOSE: The purpose of the study was to evaluate whether number and size of antral follicles can predict the outcome of in vitro fertilization-embryo transfer. METHODS: A total of 113 patients were prospectively included into this study. After 19 days of down-regulation, number and size of follicles were determined by using recent three-dimensional transvaginal ultrasound technology. Before application of gonadotropin, all follicles had been defined as antral follicles. According to size, antral follicles were categorized into four different groups: group I included antral follicles < 5 mm, group II follicles 5 10 mm; group III 11-20 mm; and group IV > 20 mm. Pregnant and non-pregnant patients were compared in terms of their number of antral follicles of group I IV. These four groups were then compared regarding implantation rate, number of retrieved oocytes, endometrium thickness, and age. RESULTS: Pregnant patients showed an significant higher number of follicles with the size between 5 and 10 mm (P = 0.04). A significant correlation was found between number of retrieved oocytes and antral follicle size of 5-10 mm (P = 0.0001). Antral follicles with a diameter between 5 and 10 mm decreased significantly with age (P = 0.008). In group III and IV, a significant correlation was found between antral follicle size (P = 0.016) and serum estradiol level after gonadotropin-releasing hormone agonist down-regulation (P = 0.011). CONCLUSIONS: We demonstrated that patients with a higher number of follicles between 5 and 10 mm showed a significantly higher pregnancy rate, whereas patients with a dominant number of antral follicles > 11 mm have a higher cancellation rate due to ovarian low response. PMID- 11042828 TI - Comparison between two hCG-to-oocyte aspiration intervals on the outcome of in vitro fertilization. AB - PURPOSE: To determine whether there was any difference in the outcome of in vitro fertilization when retrieval of oocytes was done 34 hr (group A) or 38 hr (group B) after hCG injection. METHODS: A total of 170 patients with tubal failure were randomized into either group A (83 patients) or group B (87 patients). They underwent in vitro fertilization according to described protocols and were compared with regard to the frequency of spontaneous ovulation, number of oocytes retrieved, oocyte cumulus complex quality, embryo quality, and implantation and pregnancy rates. RESULTS: There was no significant difference for any of the parameters tested for in group A and group B. CONCLUSIONS: HCG can be administered at any time within the time interval of 34 to 38 hr before retrieval of oocytes without affecting the results of in vitro fertilization. PMID- 11042829 TI - Influence of zona pellucida thickness of human embryos on clinical pregnancy outcome following in vitro fertilization treatment. AB - PURPOSE: To evaluate the correlation between the degree of zona pellucida thickness variation (ZPTV) of transferred embryos with identical morphologies and subsequent clinical pregnancy rates during 141 intracytoplasmic sperm injection (ICSI) treatment cycles. METHODS: A total of 141 women participating in the study group comprising mostly male factor infertilities and some repeat in vitro fertilization (IVF) failures were transferred, 2 days after ovum pickup, with two to three embryos with identical grades created by fertilization with ICSI. All selected embryos were subjected to zona pellucida thickness measurements immediately prior to transfer using a computerized embryo measurement program from videocinematography recordings. RESULTS: A total of 326 identical-grade transferred pre-embryos resulted in 70 clinical pregnancies with live-born implantation rate of 27.6%. A highly significant correlation was observed between ZPTV of transferred embryos and the IVF outcome with 77.1% and 83.64% of the clinical pregnancies resulting from transferred embryos with ZPTV values greater than 20 and 25, respectively. The mean ZPTV values for 70 conceptual cycles and 62 nonconceptual cycles were 28 +/- 6.43 and 17.85 +/- 8.11, respectively. No significant correlation between ZP thickness and number of blastomeres in the transferred embryos was evident, though embryos with better scores had significantly thinner zonae and higher ZPTV values. Though average zona thickness of embryos declined with age, the mean ZPTV value for women less than 30 years old was significantly higher (25.84 +/- 8.57) as compared with those from women older than 35 years (20.72 +/- 8.45). CONCLUSIONS: The degree of ZPTV of the transferred embryos exhibits a strong correlation with clinical pregnancy outcome following IVF treatment. This potentially reliable indicator of IVF success rate could be used as a criteria for embryo selection during clinical transfers. PMID- 11042830 TI - Power Doppler endometrial evaluation as a method for the prognosis of embryo implantation in an ICSI program. AB - PURPOSE: The objective of the present study was to evaluate power Doppler of the endometrium as a parameter for the prognosis of embryo implantation in patients who underwent intracytoplasmic sperm injection (ICSI). METHODS: The power Doppler was performed on a transverse section at the level of the uterine fundus on the day of human chorionic gonadotropin in 185 patients who submitted to ovarian stimulation for ICSI. The endometrium was divided into four equal quadrants and classified as grade I, II, III, or IV according to the visualization of the power Doppler in the quadrants. The color Doppler signal was considered to be positive when it reached at least the basal layer of the endometrium. RESULTS: Age, number of days of stimulation, number of follicles > or = 16 mm, number of oocytes in metaphase II retrieved, and fertilization rate did not differ patients with the four different types of endometrial grades. Endometrial thickness and the pulsatility index of uterine artery also were similar for the four grades. The rate of embryo implantation also did not differ significantly (P = 0.53) among groups: grade I = 10%; grade II = 11.6%; grade III = 15.4%; grade IV = 10.5%. The pregnancy rates were grade I = 25%; grade II = 29.7%; grade III = 37.5%; grade IV = 23.8% (P = 0.44). CONCLUSIONS: Our data demonstrate that isolated evaluation of endometrial vascularization with power Doppler is not an important factor for the prediction of pregnancy in an ICSI program. PMID- 11042832 TI - The effects of brief gamete co-incubation in human in vitro fertilization. AB - PURPOSE: To investigate whether very short exposure of mature oocytes to sperm in vitro may affect the fertilization rates, embryo cleavage rates, and embryo quality between sibling oocytes in the same patient. METHODS: Sibling oocytes of the same patient from 23 oocyte collection cycles were randomly allocated to the study group, with a 1-hr or 3-hr sperm-oocyte incubation, or the control group with the standard overnight gamete co-incubation. The fertilization rates, cleavage rates, and subsequent embryo quality were evaluated. RESULTS: Our results showed no statistically significant differences in fertilization rates, embryo cleavage rates, and quality of the embryos between the study group and the control group. CONCLUSIONS: Since the present study showed that long exposure of the oocyte to sperm has no advantage over short exposure, we prefer shortening the oocyte-sperm incubation period for reducing the negative effect induced by nonphysiologically high concentrations of spermatozoa. PMID- 11042833 TI - Blastocyst-ET and monozygotic twinning. AB - PURPOSE: To examine the rate of monozygotic twinning associated with blastocyst transfer using commercially available, cell-free culture systems with unmanipulated blastocysts. METHODS: A retrospective analysis was conducted in multiple private and academic infertility centers throughout the United States, of 199 pregnant patients following in vitro fertilization (IVF) blastocyst embryo transfer (ET). Human embryos obtained through standard IVF stimulation protocols were cultured in commercially available, cell-free media systems and transferred as blastocysts. The main outcome measure was the rate of monozygotic twinning. RESULTS: A total of 199 blastocyst-ET pregnancies were achieved during the study period at the fertility centers examined. Monozygotic twinning was noted in 10/199 (5%) of these pregnancies. All were monochorionic diamnionic. CONCLUSIONS: Monozygotic twinning previously has been reported following IVF, especially in relation to assisted hatching. While blastocyst transfer has been available for many years using coculture, there have been no published multicenter reports of monozygotic twinning associated with unmanipulated blastocysts. In a multicenter analysis, a definite increase in monozygotic twinning was seen following blastocyst-ET. We believe this phenomenon is real and that this information should be considered when counseling patients for treatment. PMID- 11042831 TI - The influence of supernumerary embryos on the clinical outcome of IVF cycles. AB - PURPOSE: To assess the influence of the presence of quality supernumerary embryos on the clinical outcome and risk of multiple conception in patients having their first in vitro fertilization (IVF) cycle. METHODS: Retrospective cohort study of 1448 women having their first IVF treatment cycle who received 4004 embryos where at least six embryos were available for transfer treated in an Assisted Conception Unit based in a large teaching hospital. RESULTS: The replacement of three rather than two embryos to women under 35 years who had good-quality supernumerary embryos resulted in a higher twin (12.5 vs. 11.9%) and triplet birth rates (2.1 vs. 0%), without significantly improving the clinical pregnancy (50.5 vs. 45.2%) or total live birth rates (38.9 vs. 35.7%). In the absence of quality spare embryos, these women who had three rather than two embryos replaced had a significantly higher clinical pregnancy rate (39.3 vs. 28.8%; P = 0.04), total live birth (32.7 vs. 19.4%; P = 0.02) and singleton birth rate per cycle (20.8 vs. 14.4%; P = 0.04), without significantly influencing the multiple birth rate. In women over 35 years, the replacement of three instead of two embryos in the presence or absence of quality supernumerary embryos led to a significant improvement in clinical outcome, without being associated with a concurrent increase in the multiple birth rate. Women in both age groups who had either two or three embryos replaced in the presence of quality supernumerary embryos had a notably better clinical outcome compared with their counterparts who had the same number of embryos replaced, but with no quality embryos to spare. CONCLUSIONS: The presence of good-quality supernumerary embryos can be used as a reference to determine the optimal number of embryos to transfer and as an indicator of the probability of success of an individual couple in a given cycle. Optimal pregnancy rates and simultaneous reduction of multiple gestation can be achieved with a flexible embryo replacement policy that is based on embryo quality, maternal age, and the presence or absence of surplus quality embryos. PMID- 11042834 TI - Effect of different concentrations of recombinant leukemia inhibitory factor on different development stage of mouse embryo in vitro. AB - PURPOSE: To assess the influence of different concentrations of recombinant human leukemia inhibitory factor (LIF) on the in vitro development of mouse embryos. METHODS: The 2- to 4-cell embryos of CB6F1 mice were cultured in the human tubal fluid (HTF) media containing different concentrations of LIF. Mouse embryos were divided into seven groups: (1) HTF; (2) 1500 IU/ml LIF; (3) 1000 IU/ml LIF; (4) 750 IU/ml LIF; (5) 500 IU/ml LIF; (6) 250 IU/ml LIF; (7) 125 IU/ml LIF. The embryonic numbers of different stages including 5-8 cell, 9-16 cell, morula, blastocyst, and hatching blastocyst were recorded. RESULTS: The percentage of early embryo stage (2-cell embryos to 6- to 16-cell stages) in all groups were nonsignificantly different. There were higher formation rates of preimplantation embryos (morula to hatching blastocyst) in groups 2, 3, 4, and 5 than in groups 1, 6 and 7. CONCLUSIONS: LIF has positive effects on preimplantation embryo development and has nonsignificant influence on the early embryo development. The lowest concentration of LIF which could provide the optimal embryo development is 500 IU/ml. PMID- 11042835 TI - Gay and lesbian couples in therapy: perspectives for the contemporary family therapist. AB - This paper outlines the major concerns of gay and lesbian couples who seek therapy. Presenting problems are classified as either internal to the relationship or as external (contextual) ones that reflect the influence of oppressive cultural and gender biases. Throughout the article, distinctive therapy methods are described that address the unique concerns of lesbian and gay couples, with special sensitivity to heterosexist and homophobic bias. PMID- 11042836 TI - Therapy with lesbian and gay parents and their children. AB - This article explores some of the social and clinical issues facing the many different kinds of gay and lesbian families that are becoming increasingly visible in the United States. Research findings are discussed that dispel popularly held myths and stereotypes concerning these families, gays and lesbians as parents, and their children. Clinical vignettes are presented to illustrate issues often encountered in the consulting room, some unique to gay and lesbian families and some common to all families. PMID- 11042837 TI - Generating stories of resilience: helping gay and lesbian youth and their families. AB - Lesbian, gay, and bisexual youth have often been forgotten in the provision of care to families. Not only are 10%-12% of all youth lesbian, gay, or bisexual, they all have families of origin and many also have "families of choice" that are dramatically increasing the numbers of persons who are directly affected. These youth can suffer disqualification ranging from internalized poor esteem and suicidality to physical danger, often at the hands of those very persons who are expected to care for them--families, educators, and health care providers. This article examines both how homophobia and heterosexism are both manifest and recovered from using case examples and offering therapeutic suggestions to clinicians. PMID- 11042838 TI - Straight therapists working with lesbians and gays in family therapy. AB - This article uses a cultural literacy model to sensitize straight marital and family therapists (MFTs) to work with gays, lesbians, and their families. While most MFTs number gays and lesbians among their clients, differences in sexual orientation between therapist and clients are often insufficiently addressed, closing off therapeutic possibilities. Marital and family therapists are asked to systematically assess homophobic and heterosexist assumptions in both personal attitudes and professional theory and practice and to educate themselves about gay culture and family life. The role of disclosure, trust, and collaborative meaning making in creating a therapeutic relationship that is culturally sensitive, clinically effective, and ethically responsible is examined. PMID- 11042839 TI - Gender in lesbian relationships: cultural, feminist, and constructionist reflections. PMID- 11042840 TI - Clients' perceptions of pivotal moments in couples therapy: a qualitative study of change in therapy. AB - Few qualitative research studies have been conducted on change processes in couples therapy, and even fewer have focused on the clients' perceptions of change processes. In this grounded theory qualitative study on marital therapy, clients' perceptions and experiences of pivotal moments are identified. Analysis of transcripts of therapy sessions, postsession questionnaires, and two posttherapy interviews with each couple revealed that clients did identify specific therapy events or discourses as pivotal. Reports of pivotal moments tended to be highly individualized accounts, with little overlap spouses and little overlap between therapist and client identification of pivotal moments. Pivotal moments tended to occur during discussions of topics that were presenting problems for the couple but, typically, after repeated discussions of the same topic. Other findings and implications are discussed. PMID- 11042841 TI - Toward a developmental family therapy: the clinical utility of research on adolescence. AB - Tremendous advances have been made in our understanding of the intrapersonal, interpersonal, familial, and contextual characteristics and processes that contribute to adaptive as well as maladaptive developmental outcomes with high risk and clinically referred adolescents. This empirical knowledge base offers clinically rich opportunities for systematic treatment development. An important step in this process is distinguishing which research findings in basic science areas such as developmental psychology and developmental psychopathology might have clinical relevance. Toward this goal, we review relevant but selective research in areas that are central to clinical work with adolescents (parent adolescent relationship, biological aspects, and affect and cognition), and we offer examples of how basic research in these areas can inform treatment. PMID- 11042842 TI - Hierarchical relationship development: parents and children. AB - Child growth and development occur in hierarchical relationships. In our attempt to work more collaboratively, family therapists have neglected to work toward developing theory that guides our work in such hierarchical relationships. This article describes a method for understanding child and relational growth. It builds on Wynne's (1984) epigenetic model of relational systems by integrating his model of family development with Bateson's (1958) concepts of complementarity and symmetry. Wynne's model defines a process for understanding the stages of relationship development, and Bateson articulates the process of change within a relationship. Utilizing principles from both theorists, this paper proposes that parent-child complementary relationships mature through Wynne's developmental stages via symmetrical struggles, and that these symmetrical struggles are necessary ingredients in the development of the relationships as well as of the individuals within those relationships. PMID- 11042843 TI - Supervising self-supervision: constructive inquiry and embedded narratives in case consultation. AB - The development of self-supervision has been suggested as a universal supervisory goal. However, the principle of isomorphism suggests that supervisors from different orientations will view this practice quite differently. This article elaborates an approach to self-supervision that is consistent with constructive therapies, though the intention is to provide flexible guidelines that can accommodate a range of supervisory activities. Case consultation is viewed as an "embedded narrative" involving the case story, the therapist story, and the supervision story. A process of "constructive inquiry" is used to connect these stories in order to construct the identity of a self-sustaining therapist. This process requires a conceptual shift from supervising practice to supervising self supervision. PMID- 11042844 TI - A test of the Circumplex Model of Marital and Family Systems using the Clinical Rating Scale. AB - Most studies of the Olson Circumplex Model of Marital and Family Systems have utilized a version of the Family Adaptability and Cohesion Evaluation Scales (FACES). Because FACES does not appear to operationalize the curvilinear dimension of the Circumplex Model, researchers have been pessimistic about the model's validity. However, the Clinical Rating Scale (CRS) has received some support as a curvilinear measure of the Circumplex Model. Therefore, we used the CRS rather than FACES to test the validity of the Circumplex Model hypotheses. Using a structural equation-modeling analytical approach, we found support for the hypotheses pertaining to the effects of cohesion and communication on family functioning. However, we found no support for the hypotheses pertaining to the concept of adaptability. We discuss these results in the context of previous studies of the Circumplex Model using FACES. Based on the collective findings, we propose a preliminary reformulation of the Circumplex Model. PMID- 11042845 TI - Clinical assessment using the Clinical Rating Scale: Thomas and Olson revisited. AB - Does the Clinical Rating Scale retain its validity when it is used, not by researchers, but by psychotherapists in their clinical practice? Students in a marital and family therapy training center routinely used the Clinical Rating Scale as part of the intake process. Although they had modest training in its use, confirmatory factor analysis indicated that they produced data that provided a reasonable approximation of the underlying factor structure. Therefore, although primarily considered a research instrument, the Clinical Rating Scale may have a role in clinical assessment and treatment planning. PMID- 11042846 TI - [Gene networks]. PMID- 11042847 TI - [Computer analysis of signals of regulation in bacterial genomes. Attenuators of operons of aromatic amino acids metabolism]. PMID- 11042848 TI - [MIR--family of repeats common for vertebrate genomes]. PMID- 11042849 TI - [The yeast genome and the first steps toward the postgenomic era]. PMID- 11042850 TI - [Overlapping genes in bacterial and bacteriophage genomes]. PMID- 11042851 TI - [Molecular mechanisms of early neurogenesis in vertebrates]. PMID- 11042852 TI - [From genomics to proteomics]. PMID- 11042853 TI - [Novel strategies of cloning identical and distinct DNA sequences from complex genomes]. PMID- 11042854 TI - [Alternative promoters and RNA processing in expression of the eukaryotic genome]. PMID- 11042855 TI - [Endogenous retroviruses: possible role in human cell function]. PMID- 11042856 TI - [Genomic imprinting and human hereditary pathology]. PMID- 11042857 TI - [A search for multiple sclerosis susceptibility genes ]. PMID- 11042858 TI - [DNA-diagnostic in oncology]. PMID- 11042859 TI - [Molecular medicine: molecular diagnostic, preventive medicine and gene therapy]. PMID- 11042860 TI - [Allele polymorphism of the dopamine transporter gene in group of patients with endogenous psychotic diseases: association with pathological syndromes]. PMID- 11042862 TI - [Genomics in 2000. Comments from Human Genome Meeting 2000]. PMID- 11042861 TI - [Molecular phylogeny of Chironomus genus based on nucleotide sequences analysis of two nuclear genes, ssp160 and globin 2b]. PMID- 11042863 TI - [Community-integrated acute psychiatric care. Options and limitations]. AB - The shift of psychiatric care from the hospital to the community has been accompanied by a reduction of hospital beds and shortened durations of inpatient treatment, but also by an increase in admissions. This evolution may be largely attributed to the prime focus of community mental health institutions on rehabilitation. The continued implementation of reforms in psychiatric care is contingent upon effectively halting the "revolving door phenomenon" by incorporating community-integrated treatment approaches into the care of acutely ill patients. Since the mid-1960s, a series of studies have established the efficacy of two community-integrated modalities for the treatment of acute psychiatric illness, i.e. home-based and day hospital treatment. In general, these approaches not only seem to be as effective as inpatient care for certain groups of patients but also reduce their need of hospitalisation, thereby contributing towards a cost effective, comprehensive psychiatric care system. PMID- 11042864 TI - [Ethical aspects of clinical neuroscience]. AB - This paper deals with ethical implications of neuroscientific research on patients as well as with the application of its results in diagnosis and treatment for brain diseases, in which a considerable demand for research exists due to their high frequency, long duration, disabling consequences, and unsatisfactory or nonexistent treatment possibilities. Such indispensable research on patients calls forth the basic ethical tension between respect of autonomy and dignity of the sick individual (as well as the avoidance of somatic and psychic risks and burdens) and the ethically justified demand for flawless research in recognizing, preventing, reducing, or eliminating disability and suffering caused by disease. The demand for research today also results from the increasing orientation of insurance companies towards scientifically proven evidence of the efficacy and safety of medical interventions: "evidence-based medicine." This is illustrated by 3 examples: (1) use of fetal brain tissue/cells from planned abortions in patients in therapy-resistant final stages of Parkinson's disease and the effects of neurotransplantation on the recipients, (2) research with demented patients incompetent to give informed consent, and (3) predictive (presymptomatic, prenatal) testing in the genetic counseling of individuals from families with Huntington's chorea. We conclude that adherence to high ethical standards is of inestimable significance, not only for those participating in research but also for public acceptance of that research. This is particularly valid in Germany, where nonobservance and the undermining of ethical principles was grossly practiced in the first half of this century on the slippery descent into the abyss. Therefore, continued scrutiny with the increasing variety of ethical problems in medical research is demanded. However, this will be achieved not by taboos and prohibitive regulations but only through open discussion between scientists, particularly probands in research, patients' relatives, and the public. This leads to four demands: listening honestly and openly, both in single cases and in the public sector, training in recognition and consideration of ethical problems, and reducing unfavorable conditions such as complex bureaucratic regulations, negative public views, overzealous efficiency, and insufficient time. Sufficient numbers of qualified personnel are needed who are trained in listening and who will have the time to do so. PMID- 11042865 TI - [Mental competence and informed consent. Clinical practice and ethical analysis]. AB - The doctrine of informed consent has become a legal and ethical standard in psychiatry today. However, ethical problems arise if patients lose the capacity to give informed consent due to their psychiatric disorders. Particularly in the field of psychiatry, the assessment of competence of informed consent to medical treatment and participation in clinical trials is a controversial issue. New empirical data suggest that a high percentage of psychiatric patients are incompetent according to defined standards for assessing their capacity to make treatment decisions. Assessing competence according to a sliding scale integrates the ethical principles of autonomy and beneficence and provides help in assessing competence in clinical practice. PMID- 11042866 TI - [Acute effects of alcohol and chronic alcoholism as causes of violent crime]. AB - To study the influence of alcohol and psychosocial variables on delinquent behavior, we coded data from the psychiatric evaluation of 254 defendants using a standardized score sheet, analyzing correlations between acute intoxication at the time of the crime (ICD 10:F10.0), diagnosis of alcohol dependency according to ICD 10 (F10.2), psycho-biographical variables, criminal history, and parameters relating to the index offence. We found that 64.6% of all defendants studied were intoxicated when committing the crime and 25.6% suffered from alcohol dependency. Alcohol intoxication correlated to occurrence of violent crime, cruelty in committing the index offence, and earlier convictions. Logistic regression, with demographic and psychosocial variables entered as covariables, revealed acute alcohol intoxication but not alcohol dependency as a predictor of violent crime (odds ratio 2.3, P = 0.02). Alcohol intoxication and dependency were also independent predictors of earlier convictions (intoxication, odds ratio 4.4, P = 0.0001; dependency, odds ratio 3.6, P = 0.003). Our findings support the hypothesis that acute alcohol intoxication, not dependency, influences violent crime in a direct manner. However, alcohol dependency predicts criminal recidivism. PMID- 11042867 TI - [Detoxification of poly-substance abusers with buprenorphine. Effects on affect, anxiety, and withdrawal symptoms]. AB - We used an open-labeled, 21-day inpatient detoxification treatment to compare the short-term effects of a 10-day buprenorphine plus 19-day carbamazepine regimen (n = 15) to a 14-day oxazepam plus 19-day carbamazepine regimen (n = 12) during rapid detoxification from opioids and other abused drugs. Somatic and psychopathological changes were assessed using the following rating scales: ASI, HAMD, SCL-90-R, and SOWS. Eighteen of 27 patients (67%) completed the study. Four dropouts (27%) were treated with buprenorphine/carbamazepine (BPN/CBZ) and the other five dropouts (42%) were treated with oxazepam/carbamazepine (OXA/CBZ). Repeated measures analysis of variance showed that SOWS scores were significantly less pronounced with BPN-CBZ than with OXA/CBZ. On the first day of admission, no significant difference in HAMD scores was detected (BPN/CBZ 11.6, BPN/CBZ 1.0). On day 14, HAMD was significantly less pronounced in BPN/CBZ (3.0) than in OXA/CBZ (6.1). BPN/CBZ showed a significant improvement in the ASI score on days 7 and 14 compared with OXA/CBZ. Three of nine items of the SCL-90-R showed a trend toward less pronounced outcome in BPN-CBZ. No severe side effects occurred during treatment in either group. The buprenorphine/carbamazepine regimen provided significantly more effective relief from affect disturbances and withdrawal syndromes than the oxazepam/carbamazepine regimen. The pharmacological basis of these effects of buprenorphine (kappa-antagonism activity,mu-agonism activity) are discussed. PMID- 11042869 TI - [Psychiatric and somatic morbidity of patients with suspected multiple chemical sensitivity syndrome (MCS)]. AB - Multiple chemical sensitivity (MCS) or idiopathic environmental intolerance (IEI) is understood as an acquired disorder with multiple recurrent symptoms that cannot be traced to any well-known medical or psychiatric condition and is associated with diverse environmental influences that are well tolerated by the majority of people. In a prospective study, we investigated 120 consecutive patients admitted a university-based outpatient department for environmental medicine during 1 year. Apart from routine medical examination and special toxicological diagnostic procedures, a structured clinical interview for DSM-IV psychiatric disorders was performed with every patient. At least one psychiatric diagnosis was found in 100 patients. The diagnostic criteria for somatoform disorders were filled by 53 patients. We found lifetime or current affective disorders in 39 patients, anxiety disorders in 29, and substance dependency or abuse in 25. In 16 patients, personality disorders were diagnosed. Nine suffered from psychotic disorders. This is the largest prospective study with standardized psychiatric diagnostic methods concerning psychiatric morbidity and MCS. The data show that many patients with environmental health problems obviously suffer from somatoform disorders but also from other, well-known psychiatric conditions. PMID- 11042868 TI - [Postpartum risk factors in the development of children born to opiate-addicted mothers; comparison between mothers with and without methadone substitution]. AB - In a retrospective case control study at the University of Frankfurt, Germany, 101 babies born to opiate-addicted mothers were identified from birth charts from 1988 to 1995. After birth, they developed a withdrawal syndrome (neonatal abstinence syndrome). Fifty control infants and their mothers were selected from neonatal wards. The group of opiate-exposed babies was subdivided into a group born to mothers without methadone treatment (n = 48) and a group born to mothers who were enrolled in a methadone program (n = 51). The methadone infants had a significantly higher mean birth weight (2822 g) than children in the group without methadone (2471 g). The abstinence syndrome was much more intense in the methadone group (convulsions 47.1%) than in heroin-exposed babies without methadone treatment (convulsions 27.1%). Women in methadone maintenance programs lived in more stable socioeconomic conditions than opiate-addicted women without methadone substitution. Moreover, they cared significantly better for their babies: 81.3% of the methadone mothers visited their children on a regular basis and 90.9% cared adequately. The data emphasize the need in future research to look more closely at the role of methadone treatment programs in the development of opiate-exposed babies. PMID- 11042870 TI - [Antagonist-induced opiate detoxification. Report on clinical experience as well as on short and intermediate range course]. AB - In a special inpatient unit for detoxification treatment of illicit drugs, antagonist-induced opiate detoxification was studied in five nonselected inpatients with polytoxicomanic abuse. The purpose was to evaluate the feasibility of this detoxification method and its impact on further reaction to treatment. During rapid detoxification under general anesthesia in an intensive care unit, no complications occurred. Withdrawal symptoms were observed in all patients over several days. During the inpatient period, no patient could be motivated to take part in a longer rehabilitation therapy. Most patients were discharged prematurely on their own demand and none made use of the rehabilitation program offered to them. All patients relapsed after relatively short times and three out of five presented for a new detoxification treatment. PMID- 11042871 TI - [Fundamentals of scientifically based ethics in the works of Eugen Bleuler]. AB - The name Eugen Bleuler is almost exclusively linked with matters concerning the nosology of schizophrenia, general psychopathology, and the relationship of clinical psychiatry with psychoanalysis. His bibliography lists various works dealing with contemporary themes about legislation, the fight against alcoholism, and, for the most part from later years, with themes about general psychology. In this article, material from Bleuler's texts are reviewed, including some which have been inaccessible up to now, which either allow interpretative conclusions on ethical grounds or have this explicitly as a topic. In particular, the analysis focuses on Bleuler's work "The Scientific Fundamentals of Ethics", published in 1939. Written in a time widely viewed as disorientated, the author coherently and systemically comments on the issue of a 'new' ethic. The anti religious and anti-philosophical positions already demonstrable in some of his earlier works will be shown. According to his conception of a scientifically based ethic, the idea of social suitability is of utmost importance and also recognisable in the animal world as a general principle of nature. Bleuler perceives the ethical 'instinct' as inherent; its absence characterises the image of an 'moral idiot', which was already a theme in his earlier works. His statements about matters concerning euthanasia are presented and, furthermore, it will be attempted to construct from his texts an underlying global view. Concerning these ethical issues, it also can be shown once more that the elderly Bleuler was hardly influenced by psychoanalytical perspectives. PMID- 11042872 TI - [Substance P receptor antagonists--a new antidepressive and anxiolytic mechanism?]. AB - Preclinical investigations suggest that the neuropeptide substance P might be involved in the etiopathology of pain, depression, and anxiety. In a recent study, the substance P receptor antagonist MK-869 showed antidepressant and anxiolytic activity in depressed outpatients which was comparable to a standard SSRI. The MK-869 was well tolerated. Although these findings are promising, further studies are necessary to prove the hypothesis that substance P receptor antagonists represent a new class of antidepressants or anxiolytics. Respective studies are currently underway. PMID- 11042873 TI - [Plant potential for detoxification (review)]. AB - Data on the uptake, excretion, and biodegradation of organic xenobiotics by plants are reviewed. Detoxification pathways operating in plants and their role in remediation of biosphere are described. Structure-, concentration, and time dependent effects of xenobiotics on the ultrastructural organization of cells are analyzed. PMID- 11042874 TI - [Characteristics and use of protein hydrolysates (review)]. AB - Properties of protein hydrolysates and possible uses of these substances in research and various branches of industry are considered. The main problem discussed in this paper is the relationship between the degree of protein conversion and characteristics (structural-functional and physicochemical) of hydrolysates. PMID- 11042875 TI - [Purification and characteristic of endo-(1--4)-beta-xylanase from Geotrichum candidum 3C]. AB - A method of purification of endo-(1-->4)-beta-xylanase (endoxylanase; EC 3.2.1.8) from the culture liquid of Geotrichum candidum 3C, grown for three days, is described. The enzyme purified 23-fold had a specific activity of 32.6 U per mg protein (yield, 14.4%). Endoxylanase was shown to be homogeneous by SDS-PAGE (molecular weight, 60 to 67 kDa). With carboxymethyl xylan as substrate, the optimum activity (determined viscosimetrically) was recorded at pH 4.0 (pI 3.4). The enzyme retained stability at pH 3.0-4.5 and 30-45 degrees C for 1 h. With xylan from beach wood, the hydrolytic activity of the enzyme (ability to saccharify the substrate) was maximum at 50 degrees C. In 72 h of exposure to 0.2 mg/ml endoxylanase, the extent of saccharification of xylans from birch wood, rye grain, and wheat straw amounted to 10, 12, and 7.7%, respectively. At 0.4 mg/ml, the extent of saccharification of birch wood xylan was as high as 20%. In the case of birch wood xylan, the initial hydrolysis products were xylooligosaccharides with degrees of polymerization in excess of four; the end products were represented by xylobiose, xylotriose, xylose, and acid xylooligosaccharides. PMID- 11042876 TI - [Trypsin inhibitor from Amaranth (Amaranthys cruentus) leaves]. AB - A protein that inhibited the proteolytic activity of trypsin was isolated from amaranth leaves (Amaranthus cruentus) by affinity chromatography on trypsin Sepharose. The inhibition was noncompetitive (with n-nitroanilide-N-alpha-benzoyl DL-arginine as substrate) and had a Ki of 11.87 x 10(-7) 7 M. The protein caused a weaker inhibitory effect on chymotrypsin, had no effect on subtilisin, displayed a molecular weight of 8 kDa, and contained no cysteine residues. PMID- 11042877 TI - [Optimization of culture media and conditions for cultivating Erwinia sp., a producer of fumarase and aspartase]. AB - Mathematical methods of experimental design were used to determine the optimal concentrations of nutrient medium components, aeration conditions, and pH providing for the maximum biomass yields, as well as fumarase and aspartase activities, during submerged cultivation of Erwinia sp. The data showed that different concentrations of carbon source (molasses) and pH of the nutrient medium were required to reach the maximum yields of fumarase and aspartase. Calculations suggested that the combination of these optimized factors would result in 3.2-, 3.4-, and 3.8-fold increases in the Erwinia sp. biomass, aspartase activity, and fumarase activity yields, respectively. The experimental data were consistent with these estimates to a 80% accuracy. PMID- 11042878 TI - [Effect of membrane-active microbial autoregulators on the growth of cultured ras transformed fibroblasts]. AB - Differential effects on proliferation of individual vs. combined administration of high- and low-molecular-weight microbial autoregulators (extracellular RNase from Bacillus subtilis and anabiosis-inducing factor d1) are reported for the first time for cultured cells of higher eukaryotes. Proliferation of ras transformed mouse fibroblasts was affected by both autoregulators dose dependently. The cytotoxic activity of individual regulators was directly related to their concentration. Unlike RNase, factor d1 (which functions as a chemical chaperone) exerted reversible effects. Studies of the effects of combined administration of the autoregulators demonstrated that pretreatment of the cells with low-dose d1 decreased the toxicity of RNase. Higher doses of d1 were required to attenuate the effects of toxic agents with more pronounced membrane tropism. The results obtained suggest that a universal system regulating the physiological activity of cells is operative in taxonomically remote organisms. The operation of the system is based on sequential changes in the structural organization and function of subcellular structures, induced by low- and high molecular-weight autoregulators. PMID- 11042879 TI - [Respiratory activity of bacteria Acinetobacter calcoaceticus TM-31 during assimilation of alkane hydrocarbons]. AB - The respiratory activity of Acinetobacter calcoaceticus TM-31 with resect to alkane hydrocarbons was studied. The dynamics of oxygen consumption by the cells while assimilating n-hexadecane was assayed by a modified technique using an oxygen electrode. The dependence of cell respiratory activity on the amount of n hexadecane within the concentration range of 0.03-0.66% was determined. It was demonstrated that the cells also displayed respiratory activity towards other medium-chain n-alkanes: hexane, octane, decane, tridecane, and heptadecane. Thus, we demonstrated the possibility of determining alkanes by measuring the respiratory activities of microorganisms. PMID- 11042880 TI - [Effect of bulk discharges of oil products on sludge microbiocenoses of purification works and their function]. AB - The effects of two bulk discharges of oil products on the microbiocenoses and functioning of bioactive sludge in purification works of Ulan-Ude city were investigated. Pollution with oil products exerted both a direct toxic effect on the microorganisms and an indirect effect via food chains. No toxic effects of oil products on sludge nitrifying bacteria were found. PMID- 11042881 TI - [An antibiotic complex produced by Streptomyces werraensis 1365T strain]. AB - An antibiotic complex comprising four components (A, B, C, and X) was extracted from a native solution and mycelium of Streptomyces werraensis 1365T. The components were purified by column and thin-layer (TLC) chromatographic procedures to study their physicochemical and biological properties. The results were used to identify the substances isolated. The preliminary data allowed us to identify the components X, A, and B as the previously described compounds undecylprodigiosin, anisomycin, and copiamycin, respectively, whereas component C is a natural compound, which probably has never been described. PMID- 11042882 TI - [Antioxidant features of fungal melanin pigments]. AB - Fungal melanin pigments were shown to display a high antioxidant activity. An increase in the number of methyl substituents in benzidine molecules of melanins obtained from micromycetes and macromycetes was accompanied by a decrease in the efficiency of inhibition of peroxidase-catalyzed oxidation. Melanins were found to have considerable gene-protecting properties. Pigments isolated from macromycetes and applied at a much lower concentration than those obtained from micromycetes prevented damage to bacteriophage-lambda DNA induced by products of peroxidase-catalyzed degradation of aminobiphenyls. PMID- 11042883 TI - [Comparative immunochemical characteristics of polyclonal and monoclonal antibodies to aflatoxin B1]. AB - Four hybrid clones (MM-(AB1)-1, MM-(AB1)-2, MM-(AB1)-3, and MM-(AB1)-4) were obtained by hybridoma technology with immunization of BALB/c mice with a BSA conjugate of aflatoxin B1 carboxymethyloxime derivative. Antibodies produced by these clones varied in the ability to recognize the aflatoxin B1 analogues. The sensitivity of enzyme immunoassay based on all monoclonal antibodies was higher compared to analysis based on polyclonal rabbit antibodies (0.1 and 0.4 ng/ml, respectively). PMID- 11042884 TI - [Composition and structure of a galactomannan macromolecules from seeds of Astragalus lehmannianus Bunge]. AB - The composition and structure of a galactomannan from seeds of Astragalus lehmannianus, an endemic legume species, is reported for the first time. The purified galactomannan (yield, 4.8%) contained 55% D-mannose and 45% D-galactose and had a molecular weight of 997.03 kDa. Its aqueous solutions were optically active and highly viscous (the specific rotation, [alpha]D, equaled +81.3 degrees; the characteristic viscosity, [eta], 868.4 ml/g). Chemical, chromatographic, and spectral (IR and 13C-NMR spectroscopy) methods were used to demonstrate that the main chain of the molecule is formed by residues of 1,4-beta D-mannopyranose, 78% of which are substituted at position 6 with single alpha-D galactopyranose. The distribution of galactose along the chain was calculated from NMR spectra: frequencies of occurrence, per pair of neighboring mannose units, of (1) two substituents, (2) one substituent, and (3) no substituents were 65.3, 31.5, and 3.2%, respectively. The specific rotation of galactomannans was shown to correlate with their content of galactose. PMID- 11042885 TI - [Effect of gibberellin and auxin on the synthesis of abscisic acid and ethylyne in buds of dormant and sprouting potato tuber]. AB - Gibberellic and beta-indolylacetic acids at concentrations of 10(-7)-10(-5) M were shown to change the hormonal status and duration of dormancy in potato tubers. Gibberellic acid shortened the dormancy and decreased the contents of abscisic acid and ethylene in apical meristems. beta-Indolylacetic acid elongated the dormancy, decreased abscisic acid production, but caused a more than tenfold increase in the production of ethylene by apical tissues. The data suggest that beta-indolylacetic acid and ethylene, as well as gibberellic and abscisic acids, are involved in the regulation of dormancy in potato tubers. PMID- 11042886 TI - [Cytoplasmic protein vaccine against bacterial hemorrhagic septicemia (aeromonosis) of fish]. AB - Water-soluble proteins from Aeromonas sobria, a causative agent of bacterial hemorrhagic septicemia of fishes, were separated into six fractions by gel chromatography on Sephadex G-100. Injections of fraction II (67 kDa) provided the highest protection of carps against the disease. Injections of proteins contained in fraction II caused stronger effects on certain biochemical parameters in the fish liver (fatty acids of phospholipids and cathepsin B and D activities) in comparison to infections of the live culture. PMID- 11042887 TI - [Lipopolysaccharides of Shigella sonnei]. AB - Immunobiological properties of native lipopolysaccharides (LPS) from virulent and avirulent strains of Shigella sonnei bacteria (LPS-V and LPS-A, respectively) were studied. In avirulent bacteria, LPS-V induced immunosuppressive activity specific of the virulent strain. LPS of the avirulent strain, whereas LPS-A lacked this property. Native LPS-V with immunosuppressive activity were isolated from the virulent strain by and immune affinity method. Treatment of LPS-V with phenol or TCA abolished its activity and converted it into the LPS-A form. The data showed that LPS-A can be converted back to the LPS-V form by redox treatment. This approach seems to be promising for activating LPS extracted from cells with TCA or a water-phenol mixture. PMID- 11042888 TI - [In Process Citation] PMID- 11042889 TI - [Severe combined immunodefiency disease (SCID) in the Arabian horse]. AB - Severe-Combined-Immunodeficiency-Disease (SCID) is discussed with special reference to its pathogenesis, clinical symptoms, pathology, and diagnosis. The disorder has been observed in the USA, Canada, Great Britain, and Australia and is characterized by an autosomal recessive mode of inheritance. The clinical features of the disease seen in Arab foals under 46 days of age are intermittent fever, (adenoviral) pneumonia, and weight loss sometimes associated with diarrhoea. From 1998 on, the SCID gene can be detected in the Netherlands by means of DNA analysis. PMID- 11042890 TI - [Diarrhea, pup mortality and Cystoisospora species (coccidiosis)]. AB - A review of coccidiosis in dogs is given on the basis of an autopsy of a puppy from a kennel in which other puppies had diarrhoea and stunted growth. Coccidiosis especially affects puppies from kennels and leads to poor growth, diarrhoea, and even death. Its occurrence is probably underestimated. PMID- 11042891 TI - [Use of captive bolt pistol without exsanguinating the animal]. PMID- 11042892 TI - [Veterinary drugs, for whom a means and for whom a worry?]. PMID- 11042893 TI - [Response to the veterinary practice group De Grensstreek. 'You should be able to blindly trust your veterinarian']. PMID- 11042894 TI - [Aleid Lubberink, veterinarian. 'One should take into account the role of the dog in the life of the owner']. PMID- 11042895 TI - [The molecular mechanisms of quantum mediator secretion in the synapse]. AB - The modern condition of knowledge about the molecular mechanisms underlying the quantal transmitter release in the central and the peripheric synapses is analysed. The data about the synaptic vesicles types, their forming, transporting to the sites of release at the nerve endings, exo- and endocytosis processes are presented. Ultrastructural and molecular organization of active zone of nerve ending and transmitter release morphofunctional unit--secretosome, which includes synaptic vesicle, exocytosis protein complex and calcium channels, are described. The basic proteins involved in the exo- and endocytosis and their interactions during transmitter release are examined. The role of the intracellular buffer systems, calcium micro- and macrodomains in the quantal transmitter secretion are considered. The reasons of the active zones functional non-uniformity and plasticity and factors reduced transmitter release in the active zone to the single quantum are analysed. PMID- 11042896 TI - [The neuronal ensembles of the brain]. AB - The conclusion about availability of two types of neuron ensembles in the central nervous structures was made on the basis of proper experimental studies and appropriate literature data analysis. The first is presented by neurons functional cooperation (Hebbian ensembles) formed during learning process on the basis of synchronization by high-frequency oscillator activity. The second is presented by morphological-and-functional structures (Kogan's neuron ensembles) formed during neurogenesis process and genetically destined for biologically valuable patterns recognition. The control of functional state of the brain neuron networks and occurred information processes is realized by non-specific systems either restrict the neurons capability of joining in the rate of global synchronization by synchronization and desynchronization actions or form and maintain the common rhythm of activity in considerable neuron populations. The latter block single elements ability to enter into specific interactions. PMID- 11042897 TI - [The rhythmic structure of the human EEG: the current state and trends of the research]. AB - During the past decade, spectral analysis has become indispensable instrument for different kinds of EEG processing. With the development of dedicated computer system, investigation of shifts in human EEG rhythm under various conditions has improved considerably. However, it is difficult to make general conclusions from this line of research, since a large number of studies are carried out using the ambiguous experimental approaches and different methods. Present paper aims to evaluate a modern state of the art in the field of human EEG rhythmical structure investigation. The results from recent relevant articles are briefly reviewed according to the universal scheme (EEG rhythm--experimental condition--observed effect). Due to such presentation, the obtained results have been summarized and some tendencies of modern investigations have been revealed. The extension of studied frequency range of rhythmical EEG components to both high (> 35 Hz) and low (< 1 Hz) frequencies, the shift to a more detailed spectral structure analysis simultaneously with ignoring the fixed boundaries of traditional EEG rhythms, the growing attempts to reveal EEG rhythmical structure correlates of cognitive activity, and a wide utilization of dynamic approaches for the analysis of brain electrical activity are discussed in some detail. The observed data are indicate of high functional significance and perspectives of human EEG rhythmical structure investigation. PMID- 11042898 TI - [The development, phenotypic characteristics and communications of Schwann cells]. AB - The Schwann cell is one of the major cell types in vertebrate peripheral nervous system. The Schwann cell precursors are derived from the neural crest and migrate along the growing axons towards their ultimate localization. Recently obtained data regarding expression of different molecules (S100 protein, myelin proteins, low-affinity nerve growth factor receptor, GAP-43, adhesion molecules) and particularly transcription factors SCIP and Krox-20 allowed to precisely characterize lineage for myelinating and non myelinating Schwann cells and their phenotypes. The special attention to communication between neurones and Schwann cells is paid. The advance in study of the biology of Schwann cell is of great practical importance for understanding the molecular basis of neuropathies and posttraumatic peripheral nerve regeneration. Schwann cells are also considered as intriguously perspective candidates for transplantation into CNS to stimulate regeneration of axons in brain and spinal cord. PMID- 11042899 TI - [Ventricular fibrillation: the current methods for analysing the degree of irregularity of the process]. AB - Ventricular fibrillation has traditionally been described as "chaotic" and in recent years there has been discussions that fibrillation may be an instance of deterministic chaos in the context of nonlinear dynamical systems theory. The current paper summarizes modern methods of mathematical analysis of the degree of electrical irregularities of the heart during VF. The traditional methods of Fourier analysis of electrocardiographic data as well as concepts of chaos theory -fractal dimension, entropy, reconstruction of attractors and some new methods such as spatial coherence have been considered. The results are discussed in context of mathematical models and hypothesis of mechanisms of VF. PMID- 11042900 TI - [Methods for the euthanasia of experimental animals--the ethics, esthetics and personnel safety]. AB - In this paper there are shown chemical and physical methods of euthanasia of vertebrate animals. All methods are divided into three categories: A) Acceptable methods of euthanasia, B) methods acceptable only for unconscious animals, C) methods that are not acceptable for euthanasia. The acceptability or non acceptability of the method is determined by ethic or aesthetic conceptions and also by conceptions of the personnel safety and the environment. There is provided a table of acceptable methods of euthanasia. Recommendations of working group of the Federation of Laboratory Animal Science Association (FELASA) were taken into consideration in this paper. The survey can be useful for the experimenters and personnel concerned with laboratory animals. PMID- 11042901 TI - [Methotrexate: a nonsurgical treatment alternative in tubal pregnancy]. PMID- 11042902 TI - [Increased efficacy of thrombolytic therapy in acute myocardial infarction by improved properties of new thrombolytic agents]. AB - Thrombolysis is a milestone in the therapy of acute myocardial infarction due to its ubiquitous availability and ease of application. The recombinant form of tissue-type plasminogen activator (rt-PA) has been proven to be superior over first generation fibrinolytic agents (streptokinase, urokinase, APSAC) with respect to efficacy and side effects and the front-loaded regimen has meanwhile become the "gold"-standard of thrombolytic therapy. Biochemically modified mutants of wild-type t-PA, e.g. r-PA (reteplase), n-PA (lanoteplase), and TNK-t PA have altered biochemical and pharmacokinetic characteristics which make them more easy to use (as double or single bolus injection) and share a theoretical efficacy benefit. This article describes these specific characteristics focussing on clot-selectivity and PAI-1 resistance of the new mutants of wild-type t-PA and their possible clinical efficiency. PMID- 11042904 TI - Gender-specific differences in the natural history, clinical features, and socioeconomic status of HIV-infected patients: experience of a treatment centre in Vienna. AB - BACKGROUND: In order to evaluate natural history, clinical features, and socioeconomic aspects in HIV-infected women, a prospective study comprising 695 HIV-infected patients was performed at our department. METHODS: Demographic data, CD4+ T-lymphocyte counts, human immunodeficiency virus-1 (HIV-1) plasma loads, AIDS indicator diseases and socioeconomic variables were recorded. Prognostic factors for survival were evaluated by univariate and multivariate analysis. Data of men and women were compared. RESULTS: The mean age of the 150 women (29.4 +/- 9.4 years) was significantly lower than that of men (32.6 +/- 9.7 years, p = 0.0002). CD4+ T-lymphocyte counts, HIV1-plasma loads, and AIDS indicator diseases did not significantly differ between men and women; the same was true for all socioeconomic variables (family status, education, professional training, employment status) except family status. No significant difference in median overall and AIDS-free survival was observed between females (2033 and 1593 days) and males (1554.5 and 1235 days, respectively, p = 0.36 and p = 0.098). Overall survival compared by age groups (< 30, 31-50, > 50 years), by risk categories (homosexuals, i.v. drug users, heterosexual contacts) and by CD4+ T-lymphocyte count (< 200, 200-500, > 500 cells/mm3), differed significantly (p < 0.001) as did AIDS-free survival. Lower age and a high CD4+ T-lymphocyte count were independently associated with the outcome in the multivariate analysis. (Overall survival/relative risk: 0.49 for age < 30 years and 2.3 for CD4+ T-lymphocyte count < 200 cells/mm3, AIDS-free survival/relative risk: 0.65 for age < 30 years and 3.3 for CD4+ T-lymphocyte count < 200 cells/mm3). HIV-1 plasma loads as a prognostic factor could not be evaluated due to the small number of patients who died or developed AIDS (2/375 and 10/375 patients, respectively). CONCLUSION: Our results showed no difference in the natural history and clinical features of HIV infection between men and women. However, pulmonary tuberculosis was associated with a significantly longer survival compared to other AIDS-defining diseases. Lower age and high CD4+ T-lymphocyte count are independent predictors for survival. With the exception of family status, socioeconomic variables showed no differences between male and female patients. PMID- 11042903 TI - Immunohistochemical expression of matrix metalloproteinases 1 and 2 (MMP-1 and MMP-2) and tissue inhibitor of metalloproteinase 2 (TIMP-2) in ruptured and non ruptured tubal ectopic pregnancies. AB - OBJECTIVE: Recent evidence suggests that matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) are crucial for trophoblast implantation in normal pregnancy. To evaluate the expression of MMP-1, MMP2, and the tissue inhibitor of MMP-2 (TIMP-2) along the invasive pathway of trophoblast in ruptured and non ruptured tubal ectopic pregnancies, we performed a retrospective immunohistochemical study. METHODS: In 15 tissue specimens of patients with ruptured (N = 7) and non-ruptured (N = 8) first trimester tubal ectopic pregnancies who underwent laparoscopic salpingectomy, immunohistochemical staining against MMP-1, MMP-2, and TIMP-2 was performed. Serial paraffin sections were photographed and digitized for a computerized quantitative image analysis. Mean percentages of positive stained areas by MMP-1, MMP-2, and TIMP-2 antibodies in the extravillous trophoblast were determined for ruptured and non-ruptured tubal ectopic pregnancies and compared. RESULTS: In our 15 tissue specimens of ectopic pregnancies MMP-1 and TIMP-2 were found to be more prominent in the immunohistochemical distribution pattern than MMP-2. However, no statistically significant difference could be detected between the mean percentages of positive stained area by MMP-1, MMP-2, and TIMP-2 antibodies in ruptured and non-ruptured tubal pregnancies. DISCUSSION: For the first time, we measured the comparative immunohistochemical expression of MMP-1, MMP-2, and TIMP-2 in ruptured and non ruptured tubal ectopic pregnancies. Although our results did not show any statistically significant difference between ruptured and non-ruptured tubal ectopic pregnancies, we conclude that MMP-1, MMP-2, and TIMP-2 are functionally involved in the highly proliferative early first part of ectopic implantation. PMID- 11042905 TI - Hybrid capture based human papillomavirus typing in cervical screening compared to cytology and histology. AB - INTRODUCTION: Cervical cancer is frequently associated with infection from various types of human papillomavirus (HPV) with high a oncogenic potential (high risk types). Commercial systems for HPV typing are available, but the question as to when HPV typing should be performed has not yet been solved. OBJECTIVES: To assess the value of HPV typing in a clinical setting in a population with opportunistic screening. STUDY DESIGN: Cytology, histology and HPV status of 593 patients from a high-risk collective were evaluated retrospectively. For HPV typing, the hybrid capture (HC) system was used. RESULTS: Infection with high risk types of HPV was associated with more severe cervical lesions. Women with PAP III or PAP IIID who were infected with high-risk HPV were at increased risk for high-grade cervical lesions (CIN III+) (p = 0.006). Conization influenced HPV status: of 63 patients who were HPV high-risk positive before conization, 4 remained positive afterwards. CONCLUSION: HC appears to be a useful system to triage women with PAP III or IIID and to detect patients with residual HPV infection after conization. However, because of high costs and no significant increase in the sensitivity of cytology, the use of HPV typing in routine cervical screening cannot be recommended in countries with opportunistic annual cytological screening. PMID- 11042907 TI - Systemic methotrexate treatment of interstitial pregnancy--magnetic resonance imaging (MRI) as a valuable tool for monitoring treatment. AB - OBJECTIVE: Interstitial pregnancy occurs in 2-4% of ectopic pregnancies and is defined as implantation of the trophoblast in the interstitial part of the tuba uterina. Therefore the term intramural pregnancy can also be found in the literature. In 20% of the cases that progress beyond 12 weeks of amenorrhea a potentially life-threatening rupture of the uterus occurs, leading to a maternal mortality rate of 2.5%. According to the literature until a few years ago diagnosis was mainly made intraoperatively, and resulted in cornual resection or hysterectomy per laparotomy. Better methods of diagnosis and treatment of interstitial pregnancy can help to decrease morbidity and mortality associated with this condition. PATIENTS: We describe two cases of interstitial pregnancies that were eventually diagnosed and also monitored by magnetic resonance imaging (MRI) after systemic methotrexate treatment. Both patients were uniparous and experienced their second spontaneous pregnancy. METHODS: Treatment consisted of four doses (50 mg/m2 body surface area) of systemic intramuscular methotrexate alternating with four doses (6 mg) of intramuscular folic acid. When beta-hCG levels were undetectable, MRI results were compared with pre-therapeutic MRI findings. RESULTS: In patients A and B, beta-hCG levels were undetectable 64 and 88 days after initiation of methotrexate treatment, while magnetic resonance imaging revealed nearly equally persisting interstitial pregnancies. They initially presented as hyperintense lesions with hypointense zones and changed into a hypointense lesion with a central hyperintense area for patient A, and a completely hyperintense lesion for patient B at the time of negative beta-hCG levels in follow-up MRI. CONCLUSION: Systemic methotrexate treatment with an intramuscular regimen is effective in the treatment of interstitial pregnancy. MRI has the ability of correct tissue differentiation and objective three dimensional measuring of interstitial pregnancy. We therefore propose this imaging modality as a valuable tool for monitoring systemic methotrexate treatment of interstitial pregnancy that should be used additionally to beta-hCG clearance curves. PMID- 11042906 TI - Upper airway findings in patients with nocturnal breathing disorders. AB - This prospective open study was performed to examine the relation between pathologic findings of the upper respiratory tract and the types of noncentral sleep disorders (rhonchopathy, obstructive sleep apnea). 312 men and 274 women aged between 35-75 years attended our outpatient department for relief of their nocturnal breathing disorder. About 1/3 of the patients suffered only of habitual rhonchopathy and 2/3 were obstructive. No findings of the head and neck examination could predict the degree of oxygen desaturation, although pathologies of the nose (73.68%), the soft palate (94.1%) and narrowing of the oro- and hypopharynx (74.19%) were very common in patients with sleep disorder. At least one pathology could be found in every patients. The more pathologies were found the higher was the risk of obstructive sleep apnoea with desaturation below 70%. Unrelated tonsillectomy (51.36%) did not prevent nocturnal breathing disorders. PMID- 11042908 TI - Giovanni Alessandro Brambilla (1728-1800) and the Imperial Infantry Regiment #22 ("Lacy"). On occasion of the bicentennial of his death. AB - Giovanni Alessandro Brambilia (1728-1800) was the Chief Surgeon of the Imperial Austrian Army and the first director of the newly founded medico-surgical academy in Vienna (1785). He died in Padua in late July 1800 (29th?), en route from his estate in Lombardy to Vienna, and the chiostro della magnolia in the Santo displays a memorial plaque commemorating his demise. On account of his merits for the education of (military) surgeons and the development of surgery in Austria, this short article shall serve to briefly recall his life and related facts. PMID- 11042909 TI - New-variant Creutzfeldt-Jacob disease. PMID- 11042910 TI - UTI antimicrobial resistance: tricky decisions ahead? PMID- 11042911 TI - Helicobacter pylori infection, intake of analgesics or anti-inflammatory medication, and personal factors in relation to dyspeptic symptoms in patients of a general practitioner. AB - BACKGROUND: Several studies have assessed the relationship between Helicobacter pylori infection and dyspeptic symptoms in highly selected patient populations and they have yielded inconsistent results. AIM: To investigate the relationship between current H. pylori infection, intake of analgesics or anti-inflammatory medication, and personal factors with dyspeptic symptoms in a large, unselected patient population of a general practitioner (GP). METHOD: Consecutive patients of a GP were invited to participate in a cross-sectional study regardless of the reason for their visit. Active infection with H. pylori was measured using the 13C-urea breath test (13C-UBT). A standardised questionnaire covering demographic, socioeconomic and lifestyle factors, and dyspeptic symptoms was completed by the patients. The number and severity of dyspeptic symptoms were quantified using a symptom score. RESULTS: Five hundred and one out of 531 eligible patients returned their questionnaires; a response rate of 94.4%. The prevalence of H. pylori infection, as indicated by a positive 13C-UBT, was 21.1% and was unrelated to dyspeptic symptoms. After adjustment for potential confounders by multiple logistic regression, a symptom score in the upper quartile of the symptom score distribution was significantly associated with female sex (odds ratio [OR] = 1.8, 95% confidence interval [CI] = 1.1 to 3.0) and intake of analgesics or anti-inflammatory drugs other than non-steroidal anti inflammatory drugs (NSAIDs) (OR = 2.3, 95% CI = 1.1 to 4.7). Older age (60 to 79 years) was associated with fewer symptoms (OR = 0.4, 95% CI = 0.2 to 0.9) when compared with the youngest age group (15 to 39 years). CONCLUSION: Female sex, younger age, and intake of analgesics or anti-inflammatory drugs other than NSAIDs, but not H. pylori infection, were independently associated with dyspeptic symptoms in this population. PMID- 11042912 TI - Patient determinants of mental health interventions in primary care. AB - BACKGROUND: A large proportion of a general practitioner's (GP's) caseload comprises patients with mental health problems. It is important to ensure that care is provided appropriately, on the basis of clinical need. It is therefore necessary to investigate the determinants of the use of mental health care in the primary care sector and, in particular, to identify any non-clinical characteristics of patients that affect the likelihood of their receiving appropriate care. AIM: To identify and compare the influence of non-clinical patient factors on GPs' acknowledgement of mental problems and on their provision of mental health care. METHOD: Cross sectional study of adults aged 16 to 65 years old (n = 802) attending one of eight practices (20 GPs in total) in inner west London. RESULTS: Multivariable analysis showed that the combination of factors that best predict GPs' acknowledgement of the presence of mental problems are general health questionnaire (GHQ) scores (odds ratio [OR] = 1.10 per unit increase in score, 95% confidence interval [CI] = 1.07 to 1.13), previous mental symptoms (OR = 7.5, 95% CI = 4.3 to 12.9), increasing age (OR = 1.03 per one-year increase, 95% CI = 1.01 to 1.04) and physical health status (OR = 0.98 per unit increase in short form-36 (SF-36) score, 95% CI = 0.96 to 1.00). Multivariable analysis showed that the combination of factors that best predict intervention (prescription for psychotropic medication; return visit to GP; referral to psychiatric inpatients/outpatients; referral to other [specified] health professionals, or social services) are previous symptoms (OR = 7.4, 95% CI = 3.8 to 14.4), white ethnic group (OR = 2.2, 95% CI 0.9 to 5.5); and not owning a property (OR = 2.1, 95% CI = 1.1 to 4.0). Life events influenced intervention only in the presence of low GHQ scores (OR = 8.1, 95% CI = 2.7 to 24.0). CONCLUSIONS: Mental problems are common in primary care and their acknowledgement is a necessary but not a sufficient condition for intervention. Our results show that GPs' decisions about mental health interventions can be influenced by non clinical patient factors, regardless of patients' clinical needs. The results suggest that current practice may not always be equitable, and point to the need for better understanding of the basis of these potential inequalities and for focused training. PMID- 11042913 TI - The evaluation of a mental health facilitator in general practice: effects on recognition, management, and outcome of mental illness. AB - BACKGROUND: Facilitation uses personal contact between the facilitator and the professional to encourage good practice and better service organisation. The model has been applied to physical illness but not to psychiatric disorders. AIM: To determine if a non-specialist facilitator can improve the recognition, management, and outcome of psychiatric illness presenting to general practitioners (GPs). METHOD: Six practices were visited over an 18-month period by a facilitator whose activities included providing guidelines and organising training initiatives. Six other practices acted as controls. Recognition (identification index of family doctors), management (psychotropic prescribing, psychological consultations with the GP, specialist mental health treatment, and the use of medical interventions and investigations), and patient outcome at four months were assessed before and after intervention. RESULTS: The mean identification index of facilitator GPs rose from 0.51 to 0.64 following intervention, while that of the control GPs fell from 0.67 to 0.59 (P = 0.046). The facilitator had no detectable effect on management or patient outcome. CONCLUSIONS: The facilitator improved recognition of psychiatric illness by GPs. Generic facilitators can be trained to take on a mental health role; however, the failure to achieve more fundamental changes in treatment and outcome implies that facilitator intervention requires development. PMID- 11042914 TI - How much does self-reported health status, measured by the SF-36, vary between electoral wards with different Jarman and Townsend scores? AB - BACKGROUND: The best way for practices to determine the health status of patients living in areas with different socioeconomic characteristics is unclear. AIMS: To see how much SF-36 health status varies between electoral wards, how much of this variation can be explained by census-derived Jarman and Townsend scores, and compare the performance of census scores with direct socioeconomic information. METHOD: A postal questionnaire survey of 3000 randomly selected 18 to 75-year olds residing in 15 electoral wards and registered with two urban practices. RESULTS: The response rate was 73%. Only two of the eight SF-36 domains were significantly associated with Jarman scores, whereas seven domains were associated with the Townsend score. Of the four socioeconomic variables derived directly from the survey, unemployment showed the weakest association, housing tenure was associated with seven domains, and car ownership and low income were associated with all eight. Income explained between 47% to 71% of the variation across the eight domains. CONCLUSION: The most accurate predictions about health status were made from direct socioeconomic information. Nonetheless, the association between Townsend score and health status was strong enough to be of practical importance. This study cautions against assuming the Jarman score of a population has a clear relationship with its health status. PMID- 11042915 TI - Management of urinary tract infection in general practice: a cost-effectiveness analysis. AB - BACKGROUND: Symptoms associated with urinary tract infection (UTI) are common in women in general practice and represent a significant burden for the National Health Service. There is considerable variation among general practitioners in the management of patients presenting with these symptoms. AIM: To identify the most appropriate patient management strategy given current information for non pregnant, adult women presenting in general practice with symptoms of uncomplicated UTI. METHOD: A decision analytic model incorporating a variety of patient management strategies was constructed using available published information and expert opinion. This model was able to provide guidance on current best practice based upon cost-effectiveness (cost per symptom-free day). RESULTS: Empiric treatment was found to be the least costly strategy available. It saved two days of symptoms per episode of UTI at a cost of 14 Pounds. The empiric-and-laboratory strategy involves an incremental cost-effectiveness ratio of 215 Pounds per symptom day averted per episode of UTI. The remaining patient management strategies are never optimal. CONCLUSION: Empiric treatment of patients presenting with symptoms of UTI was found to be cost-effective under a range of assumptions for this patient group. However, recognition of the impact of this strategy upon antibiotic resistance may lead to the dipstick strategy being considered a superior strategy overall. PMID- 11042916 TI - Low back pain in general practice: reported management and reasons for not adhering to the guidelines in The Netherlands. AB - BACKGROUND: Although guidelines for the management of low back pain have been published in the past decade, there is potential for further improvement in back pain care. AIM: To document the management of non-specific low back pain by general practitioners (GPs) in the Netherlands, to determine how this management of care is related to patient and physician factors, and to explore possible reasons for not adhering to the guidelines. METHOD: A prospective study was set up in which 57 GPs in 30 general practices completed a computerised questionnaire after each consultation for low back pain during a four-month period. RESULTS: Of 1640 back pain contacts, 1180 referred to non-specific low back pain. Diagnostic tests were ordered in 2% of first consultations and in 7% of follow-up consultations within one episode. The advice to stay active despite pain was given in 76% and 69% of these cases respectively. Patients were prescribed an analgesic in 53% and 41% of cases respectively (mainly NSAIDs [80%]). Patients were referred to a physiotherapist in 22% of first and in 50% of follow-up consultations. Older patients were physically examined less often, prescribed analgesics more often, and were told less often that staying active could benefit them. The advice to remain active was omitted more often when symptoms lasted longer. Only a small part of the variance in management was accounted for by patient characteristics or by differences between practices. CONCLUSION: The management of low back pain met the guidelines to a large extent. Management decisions were often related to characteristics in which the guidelines lack differentiation. Important reasons for non-adherence were perceived patients' preferences. Further implementation of guidelines will be difficult unless doctors' and patients' views are more explicitly known. PMID- 11042917 TI - General practitioners miss disability and anxiety as well as depression in their patients with osteoarthritis. AB - BACKGROUND: General practitioners (GPs) integrate physical, psychological, and social factors when assessing patients, particularly those with chronic diseases. Recently, the emphasis has been on assessment of depression but not of other factors. AIM: To determine functional disability, psychological morbidity, social situation, and use of health and social services in patients with osteoarthritis and examine GP knowledge of these factors. METHOD: Two hundred patients completed a validated postal questionnaire about functional disability (Health Assessment Questionnaire [HAQ]), mood (Hospital Anxiety and Depression Scale [HAD]), employment status, who they lived with, welfare benefits received, and use of health and social services. A similar questionnaire was completed by the patient's GP, including a HAQ. However, a three-point scale was used to assess depression and anxiety. RESULTS: Forty-seven per cent of patients were moderately or severely disabled (HAQ > 1). GPs underestimated functional disability: mean patient HAQ = 1.04 (95% confidence interval [CI] = 0.92-1.16), mean GP HAQ = 0.74 (95% CI = 0.65-0.83), and there was low correlation between patient and GP scores (kappa = 0.24). There was moderate prevalence of depression and high prevalence of anxiety, which the GP often did not recognise: patient depression = 8.3% (95% CI = 4.1%-12.8%), GP depression = 6.0% (95% CI = 2.4%-9.6%), kappa = 0.11; patient anxiety = 24.4% (95% CI = 17.8%-31.0%), GP anxiety = 11.9% (95% CI = 6.9% 16.9%), kappa = 0.19. Only 46% of severely disabled patients (HAQ > 2) were receiving disability welfare benefits. GPs were often unaware of welfare benefits received or the involvement of other professionals. CONCLUSION: GPs frequently lack knowledge about functional disability, social factors, and anxiety as well as depression in their patients with osteoarthritis. PMID- 11042918 TI - Towards a conceptual framework for evaluating primary care research networks. AB - We describe a conceptual framework that we have developed for evaluating primary care research networks. The framework includes objectives, process indicators, and outcome indicators. We propose the framework as a provisional model that we hope will promote further research and debate. PMID- 11042919 TI - Palliative care provided by GPs: the carer's viewpoint. AB - As most terminal and palliative care is in the community, general practitioners (GPs) have an important role to play. This study presents bereaved carers' views of the palliative care provided by GPs. It suggests that symptom control may not be optimal. PMID- 11042920 TI - What proportion of patients refuse consent to data collection from their records for research purposes? AB - In a randomised trial of the implementation of guidelines for asthma and angina, we sent questionnaires that included a request for consent to collect data from the patient's clinical records to 5069 patients in 81 general practices. Of these 3429 (67.6%) responded, of whom 335 (9.8% [95%, CI = 8.8%-10.8%]) refused consent. We conclude that consent should always be sought unless a research ethics committee has waived this requirement for pressing reasons. PMID- 11042921 TI - Copying general practitioner referral letters to patients: a study of patients' views. AB - Over the past decade there has been a significant shift towards greater involvement of patients in their health care and this has highlighted many areas relating to doctor-patient communication. One area of communication that has not been extensively researched is the referral letter between general practitioners (GPs) and their patients. This small study of patients' views suggests that patients value receiving a copy of their GP outpatient letter, appreciating greater understanding of, and involvement in, the referral process. PMID- 11042922 TI - A targeted approach to reducing maternal smoking. AB - The Government White Paper, Smoking Kills, published in December 1998, set new and more ambitious targets for reducing maternal smoking. This is despite the fact that consecutive surveys have shown that the prevalence of maternal smoking has not changed since the 1992 targets (White Paper, The Health of the Nation). Based on current literature, including the author's own research on maternal smoking, this article argues that future research and community smoking cessation interventions should: encompass not just pregnant woman but also partners and close family members; pay particular attention to young, socially disadvantaged groups; and develop and evaluate stage-dependent antenatal smoking cessation materials (tailored to the user's level of intention to quit). By adopting these measures, researchers and primary health care professionals may finally reduce infant deaths and the numerous infant and child health problems related to maternal smoking and household tobacco exposure. PMID- 11042923 TI - Primary care in the United States. PMID- 11042924 TI - Excipient E110: a cause for complaint? PMID- 11042925 TI - Randomised controlled trials in general practice. PMID- 11042926 TI - Treatment of drug users. PMID- 11042927 TI - Treatment of drug users. PMID- 11042928 TI - The Shipman inquiry. PMID- 11042929 TI - GPs' diagnosis of dementia. PMID- 11042930 TI - Who is a frequent attender? PMID- 11042931 TI - Who is a frequent attender? PMID- 11042932 TI - A better use of corticosteroids. PMID- 11042933 TI - Conflicting advice in the care of a dying patient. PMID- 11042934 TI - Key developments in endocrinology. PMID- 11042935 TI - The GP's role in growth hormone deficiency. PMID- 11042936 TI - Diabetic eye disease: how can it be prevented? PMID- 11042937 TI - Casebook: hypercalcaemia. PMID- 11042938 TI - The Alaska Blind Child Discovery project: rationale, methods and results of 4000 screenings. AB - BACKGROUND: Photoscreening allows lay persons to adapt the Enhanced Bruckner Test to preschoolers in an attempt to identify refractive amblyopia. The Alaska Blind Child Discovery (ABCD) project is charitably funded and administered. METHODS: MTI photoscreening was offered to children in rural and urban communities in southern Alaska from 1996 through June 1999. Parents answered questions concerning the child's health, family ocular history and whether the child had any eye "Warning Signs." The MTI images were interpreted by two eye doctors using a modification in MTI published guidelines. RESULTS: Out of 4000 screenings performed on 3930 children, there was an overall "not normal" interpretation of 9% and an inconclusive rate of 1%. The mean S.D. age was 3.9 2 years. Only 6% had had a prior eye exam. The average number of Polaroid pictures per screening was 1.16. Follow-up data on "not normal" results was obtained on just over 50%. The positive predictive value during the first two years was 77% but improved to 92% from 1998-1999. Affirmative answers to the questions concerning previous eye exam, child's health, siblings eye health and positive "Warning Signs" were significantly associated with "not normal" interpretations but affirmative answers about eye health of mother, father and relatives were not. Community penetrance of photoscreening to the target age-group ranged from only 5% for Anchorage to almost 100% for the Bristol Bay public health nurses. Five percent of parents of "positive" results surveyed would not have recommended screening for their friends. Equipment functioned dependably even in remote Alaska. CONCLUSION: Charitable volunteer Polaroid photoscreening detected amblyopia and significant pediatric eye disease in over 300 children during the first 3.5 years of ABCD. PMID- 11042939 TI - The detection of congenital glaucoma by photoscreen interpretation. AB - Photoscreening is designed to detect abnormalities in children's eye, particularly abnormal refractive errors, which can lead to amblyopia. An Alaska Bind Child Discovery MTI Polaroid photoscreen in one girl resulted in diagnosis and treatment of congenital glaucoma. Patients with known pediatric eye disease underwent photoscreening. Subtle non-refractive changes in photoscreen images may reveal eye disease even more serious than amblyopia. We suggest that human or computer interpretation of photoscreening images, particularly when retained, be done conscientiously with respect to the refractive state and alignment of the eyes, but also regarding other potentially serious ocular pathology. PMID- 11042940 TI - Alaska's obstetrical delivery systems: a descriptive epidemiologic study. AB - Delivery of obstetrical care in rural Alaska can be very challenging, due to remoteness, lack of medical resources and transportation difficulties. This descriptive study looks at what the current delivery systems for obstetrical care in Alaska are. Alaska's obstetrical delivery systems can be divided into three basic systems. 1) Full comprehensive obstetrical care limited only by lack of neonatal ICU capability. 2) Cesarean delivery capable, but with limited resources. 3) Low risk vaginal deliveries with no cesarean delivery capability except by transports approaching 6 hours. This study raises questions about which system is most effective for which communities. Further studies need to be undertaken to better understand how to provide effective obstetrical care in rural and bush Alaska at an acceptable risk, and at reasonable cost. PMID- 11042942 TI - 1970 preventive dentistry program. Immediate and long-term results of a control program. PMID- 11042941 TI - 1970 report on bone tuberculosis and Dr. Phil Moore's retirement. PMID- 11042943 TI - Small colony variants of Staphylococci: a link to persistent infections. AB - In prospective studies, Staphylococcus aureus small-colony variants (SCVs) have been linked to persistent and recurrent infections. SCVs are a naturally occurring subpopulation often defective in electron transport which may be identified in the microbiological laboratory as nonpigmented, nonhemolytic, slow growing pinpoint colonies after incubation on rabbit blood agar. In addition, the often relatively unstable SCVs demonstrate a number of other characteristics that are atypical for S. aureus including reduced alpha-toxin production and delayed coagulase activity. A site-directed hemB mutant with a stable SCV phenotype provided strong evidence for the link between these electron transport defective strains and persistent infections. The hemB mutant was phagocytized by cultured endothelial cells, but did not lyse these cells, because the mutant produced very little alpha-toxin. Thus, SCVs can hide within the host cell, then revert to the highly virulent rapidly growing form and lyse the host cell, once the host immune response has abated and antibiotic therapy is completed. The intracellular position shields SCVs from host defenses and decreases exposure to antibiotics. This review discusses what is known of the biology of SCVs and describes the recovery and significance of Staphylococcus SCVs in clinical specimen. PMID- 11042944 TI - [Clinical and subclinical lameness in young fattenin cattle]. AB - Time dependent changes of subclinical and clinical lameness were analysed in 493 fattening bulls in three different herds with a total number of 30,621 animals. In the majority of cases subclinical lameness were observed which were caused by metabolic acidosis or deficiencies in mineral intake (P and probably Ca). The activity of alkaline phosphatase in plasma exhibited an increase and the dry matter in bone (tubera coxae) a decrease before the appearance of clinical symptoms. Changes in bone composition were accompanied by alterations of plasma Ca and P concentrations, which finally led to clinical lameness. Hence the diagnosis was verified by simple laboratory methods as the precondition for successful treatment and prophylaxis of lameness caused by metabolic disorders. PMID- 11042945 TI - [Possibilities for standardization of ELISA for detection of Salmonella antibodies in sera and meat juices of pigs]. AB - Programmes for controlling salmonella infections in German piggeries are based on the meat-juice-ELISA conducted in various investigation centres by using different test-kits. A usual procedure for harmonization (standardisation) of results is the calculation of the percentage of antibody-concentration from field samples in relation to the extinctions of a set of control-sera with known antibody concentrations. Whether this system is still acceptable in case of using different test-kits seems to be questionable. In principle, difficulties arise by calculating field results from the regression curve of control-sera because the calculated percentages of antibodies do not represent the antibody concentration but, instead, the percentages of the extinctions measured, and secondly, because control-sera presently in use are directed against different salmonella serovars. In regard to the number of laboratories involved and because of a variety of test kits used it seems to be more adequate to include only one anti-Salmonella Typhimurium standard-serum at a given antibody concentration which is to be tested repeatedly on every test-plate. Simultaneously, further controls should include another anti-Salmonella Typhimurium and one anti-Salmonella Choleraesuis serum which should provide results similar to the Danish system which is regarded as a standard. As well, a negative serum must be included in the test and a minimum difference in extinctions between this negative serum and the standard positive control-serum should be reached to prove the validity of results from the test plate. PMID- 11042946 TI - [Technique of abdominal ultrasonography in newborn foals and normal findings]. AB - Under field conditions, the diagnosis of foal's diseases relies almost exclusively on the physical examination. As the signs of illness in the equine neonate are frequently vage and non-localizing, the diagnosis of diseases may be problematic. This often causes misinterpretations and leads to ineffective prophylaxis and treatment. The purpose of this study was to evaluate the usefulness of diagnostic ultrasonography of the foal's abdomen under field conditions to provide an optimized technique and to describe the normal findings. Diagnostic ultrasonography of the abdomen was performed after obtaining clinical history and passing the physical examination of 25 foals without signs of abdominal problems. The foals were scanned in a stable box, being restrained by three persons in semi-lateral recumbency. Usually, sedation was not necessary. The ventral abdominal wall was clipped, a generous amount of ultrasound coupling gel was applied and massaged on the skin surface. The ultrasonographic examination was carried out using a portable sector scanner ("Microimager 2000", Ausonics) with 5.0 and 7.5-MHz transducers or a combined 5.0 and 7.5-MHz transrectal linear-array scanner ("450 Enhanced", Pie Medical). Employing the 5.0 MHz sector scanner first, the abdomen was explored from caudal to cranial in left and right semi-lateral recumbency. The 7.5-MHz scanner was used to attain higher resolution of certain structures. The sector scanner turned out to be suitable under field conditions and adequate to examine the abdominal organs. The transrectal linear-array scanner also provided the most important informations, although it was difficult to maintain a good contact area of the scan head. By ultrasonography it was possible to identify the urinary bladder, kidneys, spleen, liver and part of the gastrointestinal tract. Thus, application of ultrasound could successfully be performed on newborn foals under field conditions. PMID- 11042947 TI - [Determination of breakpoints for veterinary medically relevant antibiotics for resistance assessment of veterinary pathogens]. AB - This article describes the meaning of the term clinical breakpoint. This is followed by a discussion of the parameters that need to be considered when setting valid breakpoints for active substances in veterinary medicine; in doing so we closely follow equivalent regulations and guidelines on the establishment of breakpoints in human medicine. Along with pharmacokinetic data and the results of clinical efficacy tests, susceptibility data of relevant organisms play a key role in the establishment of breakpoints. Published breakpoints are currently only available for a few modern drugs in veterinary medicine. PMID- 11042948 TI - [Influence of diagnostic tests for estimating prevalence during surveillance programs]. AB - The influence of diagnostic tests on the estimation of the prevalence and the calculation of sample sizes with respect to different sampling schemes are presented in this paper. These sampling schemes are used for the implementation of surveillance programs. Assuming "perfect tests" (i.e. sensitivity = specificity = 100%) the calculated sample sizes, e.g. for an IBR/IPV-surveillance or a paratuberculosis survey, are half of the sample sizes considering sensitivity and specificity of the diagnostic tests. However the probability not to identify infected livestocks may be 4 times higher neglecting the test characteristics and assuming perfect tests. PMID- 11042949 TI - Population-based incidence rate of invasive pneumococcal infection in children: Vancouver, 1994-1998. PMID- 11042950 TI - Non-return rates for HIV testing: results of a 3-month retrospective review at an STD clinic in 1999. PMID- 11042951 TI - The future of radiologic imaging and intervention. PMID- 11042952 TI - Intraoperative gamma probe directed rib resection. AB - Radioguided surgery has slowly gained acceptance since the first gamma probe directed sentinel node lymph node biopsy in a patient with melanoma in 1993. We describe how the intraoperative gamma probe is used to localize a rib with abnormal uptake on the bone scan in a patient with rib pain. PMID- 11042953 TI - Endovascular therapy in the treatment of head and neck lesions. AB - Recent advances in microcatheter technology, refinements in embolic agents and improvements in navigational techniques have allowed for endovascular embolization to become an important adjunct in the treatment of vascular head and neck lesions. We describe several case reports where endovascular embolization was utilized in the treatment of such lesions. PMID- 11042954 TI - Uterine fibroids: primary treatment with therapeutic embolization. AB - The use of transcatheter uterine artery embolization as a treatment for uterine fibroids represents a new approach to the management of this common problem. Early reports of uterine artery embolization as a treatment of symptomatic fibroids have indicated significant symptomatic improvement as well as reduction in the size of fibroids. We have been performing this procedure for two years at the Christiana Care Hospital. We describe two representative cases with clinical follow up on 12 of our patients and briefly review the literature on uterine artery embolization for uterine fibroids. PMID- 11042955 TI - [Statistical estimation of parameters in allometric equations]. AB - An algorithm for estimating allometric coefficients widely used in biological studies is presented. The coefficients can be estimated only when the relationship between logarithms of the approximated data meets the linearity criterion. The proposed algorithm was applied for the brain-body weight relationship in mammals and oxygen consumption rate-body weight relationship in amphibians. PMID- 11042956 TI - [Effect of the population density on growth and regeneration in the snail Achatina fulica]. AB - In the laboratory, the growth rate of the giant African snail Achatina fulica, as estimated by the weight and shell length was shown to decrease when the population density increased from 10 to 60 snails/m2 of the total terrarium area for five months. In the second experiment, when the population density increased from 48 to 193 snails/m2, the growth rate had already decreased by six weeks. In the groups with a high population density the feeding behavior was weakened, expressed by a greater amount of nonconsumed food, according to visual observations, than in the groups with lower population densities. At the population density of 10 to 60 snails/m2, the proliferative activity in the course of the optic tentacle regeneration, as expressed by the mitotic index, did not differ reliably within five months. In the second experiment, the mitotic indices at the population densities of 96 and 193 snails/m2 within 1.5 months exceeded that of 48 snails/m2. Recommendations are given concerning the population density from the viewpoint of commercial growth of the snails. It was proposed that, based on the analysis of the mechanism underlying the inhibition of feeding behavior in populations with extra high densities, one may develop a new approach to the production of chemical agents to control land snails as agricultural pests. PMID- 11042957 TI - [Comparative study of various biochemical parameters in pathogenic and nonpathogenic Aeromonas strains]. AB - We studied certain biochemical properties of virulent and avirulent strains of mobile aeromonads. The pathogenic strain featured a higher proportion of odd fatty acids in the lipids, increased protease activity, and a high concentration of 55 kDa protein. We propose that these compounds be used as pathogenic markers of these microorganisms. PMID- 11042958 TI - [Formation of flavonoid pigments during flower color development in sweet pea Lathyrus odoratus L. mutants]. AB - We studied the variability of two classes of flavonoid pigments (flavonols and anthocyanidins) in seven sweet pea mutant lines. Flavonols proved to be synthesized before anthocyanidins during the color formation of the petals. The top content of flavonols was observed at the third stage of flower development, while the anthocyanidins content peaked at the fifth stage. The pigments with the more hydroxylated B ring were synthesized at later stages of the flower development as well. The stages of flavonols and anthocyanidins biosynthesis controlled by the corresponding genes, as well as the possible genetic mechanism of a fast and complete transformation of the precursors to the final products of the biosynthetic chain, are considered. PMID- 11042959 TI - [The mobile vector method for genetic transformation of animals]. AB - Experimental data on transgenic animal production using transfected sperm are reviewed. Pathways for further improvement of the method for increasing the frequency of transgenic organisms are proposed: modification of the saccharophosphate skeleton and DNA ends; use of nucleic exchange enzymes and directed oligonucleotides; use of adhesive sequences (MAR elements), ori replication, and transposable elements within the transgenes; development of the embryos selection; the use of gene targeting elements; and the use of various DNA conformations. The promising use of spermatozoa as a noninvasive system of gene delivery in transgenesis and prenatal gene therapy are outlined. PMID- 11042960 TI - [Variations of flower coloration in garden rose spontaneous mutants]. AB - Twenty two varieties were isolated as a result of studying the world assortment of garden roses, from which many natural bud variations (sports) were selected. The genealogy of these varieties and variations of flower coloration in their sports were studied. It was shown that flower coloration may have a certain direction, depending on the origin of initial forms, i.e., on their genotype. The possible genetic causes of changes in the flower coloration in a certain predominant direction upon mutation are discussed. PMID- 11042961 TI - [Phytohormones of rhizomes of the mint of various geographic origin in its annual developmental cycle]. AB - We studied the ratio of cytokinins and abscisic acid in the rhizome tissues of two southern introduced forms of mint (GBS 1-94 and GBS 2-94) in the annual cycle of its development under the climatic conditions of the Middle Zone of Russia. The level of cytokinins exceeded that of free abscisic acid during the entire period of rhizome growth and development. Three peaks of an excess--zeatin riboside over abscisic acid were established: (a) during mass flowering, (b) after dying off terrestrial shoots, and (c) before the appearance of shoots of the next year vegetation. These peaks were most distinct in the GBS 2-94 mint. Zeatin riboside was the major identified cytokinin. The role of balance of phytohormones in rhizome tissues upon adaptation of the introduced mint forms to new climatic conditions is discussed. PMID- 11042962 TI - [Biochemical adaptation of the barley root cells to toxic substances. 1. Effect of aluminum on the phosphohydrolase activity]. AB - Acid phosphatase (AP) and two nucleotidases with a top affinity to ATP and Ca (AN1) or AMP and Mg (AN2) were found among acid phosphohydrolases of the apoplast. After 15 min aluminum chloride at 100 microM induced activity of both membrane-bound and soluble phosphohydrolases. The highest induction of the enzymes by aluminum was observed at pH 4.5. A relatively high concentration of aluminum chloride (2 mM) stimulated AN2 and inhibited AN1, while AP activity remained unaltered. We propose that they activation of membrane-bound and soluble acid phosphohydrolases is one of the protective mechanisms of barley root apoplast against the toxic effect of aluminum chloride. PMID- 11042963 TI - [Response of the algae Gymnodinium kovalevskii (Dinophyta) to exposure to synthetic detergents and distillation]. AB - The effects of synthetic detergents and combined effects of synthetic detergents and water freshening on growth characteristics of the alga Gymnodinium kovalevskii (Dinophyta) were studied. Low concentrations of synthetic detergents (0.1 and 1.0 mg/l) stimulated the algal growth. Elevated concentrations inhibited cell division, affected their motility and induced morphological changes. Contamination with synthetic detergents adversely affected the adaptation plasticity of algae with respect to salinity. PMID- 11042964 TI - [Origin of Lepidoptera fauna of the Southern Transural region]. AB - The butterfly fauna of the Southern Transural region began mainly through the migration of insects from the Urals and Kazakhstan, since the end of the Cretaceous Period to the end of Paleogen, the Transural region was covered by an epiplatform sea. As this sea was retreating, the first regions of dry land appeared, which had boundaries with Kazakhstan and the Urals. They were the first to be populated by Lepidoptera. During the Pleocene and then after the Pleistocene cooling events, insects settled generally along the valley of the Tobol River and the Turgai depression, because these territories belong to intrazonal elements. At the present time, the greatest species diversity among insects in the southern Transural area is observed specifically in the Turgai depression and in areas directly adjacent to it. This territory is mainly occupied by populations unique to the Transural regions and belonging to the following species: Mantis religiosa (praying mantis), Saga pedo, Parnassius apollo (apollo), Neolycaena rhymnus, Hyponephele lupina (oriental meadow brown), Chazara persephone (dark rockbrown), Epicallia villica (cream-spot tiger), etc. PMID- 11042965 TI - [Ecophysiological characteristics of Mytilus edulis L. living near the chalybeate spring]. AB - The White Sea mussels Mytilus edulis constantly living near chalybeate accumulate significant amounts of iron as well as zinc and manganese in soft tissues that exceed the background leve 5-10 and 2-3 times, respectively at the upper habitat limit. These mollusks feature iron particles in the blood system. It was established from chalybeate streamways appearing in chalybeate water during reflux that M. edulis tightly pressurizes its shell for this period. As a result, calcium content increases in hemolymph and cavity fluid at the expense of calcium joint of the shell. By the beginning of flux, it reaches ca. 12%; hypostracum of the shell shows specific signs of dissolution, that are manifested as brown prints of blood vessels due to their filling with iron particles. In the period of maximum drying no overhydration is observed in the mollusks and a constantly high concentration of salt is preserved in the cavity fluid. PMID- 11042966 TI - [Ratio of r- and K-forms of selection in life cycles of trematodes (Plathelminthes, Trematoda). AB - The situation is considered in a continuum of r-K-selection of hermaphroditic and parthenogenetic populations of trematodes (fish parasites) of three species: Bunodera luciopercae, Sphaerostomum bramae, and Phyllodistomum elongatum. Maritae of B. luciopercae and S. bramae are characterized by a low fertility, single-type reproduction, and long prereproductive period, and are involved in interspecific competition. On the contrary, maritae of Ph. elongatum rapidly mature, are highly fertile, repeatedly reproduce, and occur in a "competitive vacuum". Pathenitae of these trematodes combine the features of both forms of selection, which is accounted for by adaptations to the parasitic mode of life. A reproductive tactic of parthenitae has been proposed, based on the redioid or sporocystoid types. According to the community of features, B luciopercae as a species is a "K strategiest", S. bramae equally combines the features of both forms of selection, and Ph. elongatum is an "r-strategist". PMID- 11042967 TI - [Environmental activity of earthworms (Lumbricus terrestris L.) and the spatial organization of soil communities]. AB - The effect of feeding and burrowing activities of anecic earthworms (Lumbricus terrestris) on abiotic characteristics of the soil, biomass and activity of soil microorganisms, and the spatial distribution of Collembola and Lumbricidae species was studied in a Iinden forest near Moscow. The results showed that organic carbon content, nitrogen content, pH, and microbial biomass and basal respiration are considerably higher around L. terrestris burrows than in the surrounding soil. The total density of springtails near the burrows was 1.6-1.7 as high as at the control sites. The most pronounced preference for earthworm burrows was observed in the species dominating in the soils of undisturbed deciduous forests (Isotomiella minor and Isotoma notabilis). The number and biomass of epigeic and endogeic earthworms also increased significantly in the zone of L. terrestris burrows. However, some springtail (Isotoma viridis, Protaphorura cf. nemorata, Lepidocyrtus lignorum) and earthworm species (Aporrectodea rosea) did not accumulate near L. terrestris burrows and even avoided them. Thus, L. terrestris activities create a mosaic of soil microhabitats, which provides for the coexistence of different microcommunities of soil organisms. PMID- 11042968 TI - [Thermodynamic interpretation of the nitrogen status of forest gray soil agrocenoses]. AB - Thermodynamic approaches used in soil agrochemical studies allow to determine the direction of processes occurring in soils and to provide theoretically meaningful predictions of changes in ecosystems as a whole in response to various factors. Analysis of redox reactions of the NH4+ reversible NO3- equilibrium in agrocenoses of gray forest soil has shown that the additional introduction of nitrogen fertilizers is undesirable both from the economical and ecological points of view, since the dominating process occurring with the loss of nitrogen from the soil in the form of gaseous compounds is thermodynamically most preferred. PMID- 11042969 TI - [Analysis of nuclear 'trypsin-like' and 'trypsin inhibitor' coleoptile complexes at the post-embryonic stage of wheat ontogenesis]. AB - "Inhibitor-trypsin" and "trypsin-like" complexes of the nuclear matrix of actively growing wheat coleoptiles and germinated wheat embryos were isolated using affine chromatography on columns with immobilized trypsin and trypsin inhibitor, respectively. No such complexes were found after the termination of the coleoptile growth or in the air-dry quiescent wheat embryos. Electron microscopy of the "trypsin-like" complexes has shown a rosette- and bead-like structure with a dense granular layer which was absent in the "inhibitor-trypsin" complexes. Electrophoretic analysis demonstrated a range of protein molecular masses from approximately 14 to approximately 67 kDa. DNA, RNA. Carbohydrates were also found in the complexes. Immunoenzymatic analysis visualized endogenous auxin in the complexes of 24-hour germinated wheat embryos. PMID- 11042970 TI - [Effects of neurohypophysial hormones and their synthetic analogs on platelet aggregation]. AB - The effect of neurohypophysial hormones vasopressin and oxytocin as well as their synthetic analogs on platelet aggregation induced by thrombin or ADP was studied in vitro on washed rat platelets. Vasopressin and its analog DGAVP, as well as oxytocin enhanced thrombin-induced platelet aggregation, while their effect on ADP-induced aggregation was considerably lower. An oxytocin analog depotocin had a pronounced antithrombin effect indicated by a significant inhibition (by 81%) of thrombin-induced platelet aggregation and by assay for its antithrombin activity in the blood plasma. PMID- 11042971 TI - [Binding of aldosterone with cytoplasmic and brain nuclear corticosteroid receptors in rats with various types of behavior]. AB - The results are given of in vivo experiments on the binding of (3H)aldosterone to corticosteroid receptors of the cytoplasm and nuclei in the brain structure in rats with different types of behavior. Differences were established in binding (3H)aldosterone to corticosteroid receptors of the hippocampus cytoplasm in the animals with different types of behavior both under the normal conditions and within two weeks after stress. Stress was shown to affect the binding of this hormone to corticosteroid receptors of the brain cytoplasm (without hippocampus and cerebellum). No differences were found in the binding of aldosterone to corticosteroid receptors of the brain nuclei after stress. PMID- 11042972 TI - The perfect patient. PMID- 11042973 TI - Fast-curing lights. PMID- 11042974 TI - Low-dose aspirin therapy. PMID- 11042975 TI - The anti-tobacco-use bandwagon. PMID- 11042976 TI - Incomplete tooth fractures. PMID- 11042977 TI - CPR recertification. PMID- 11042979 TI - New dental therapeutics guide to debut in October. PMID- 11042978 TI - Early vs. late orthodontic treatment. PMID- 11042980 TI - Cleft lip and palate gene identified. PMID- 11042981 TI - Do you presently offer your patients tobacco-use cessation services? PMID- 11042982 TI - The dental unit waterline controversy: defusing the myths, defining the solutions. AB - BACKGROUND AND OVERVIEW: This article reviews the literature on the subject of dental unit waterline contamination. It has been expanded from the text of a lecture given at the Scientific Frontiers in Dentistry program sponsored by the National Institute for Dental and Craniofacial Research in Bethesda, Md., in July 1999. The author examines the underlying biological causes of waterline colonization by microorganisms, the evidence of potential health consequences and possible means of improving the quality of dental water. He also describes examples of devices currently marketed to improve and maintain the quality of dental treatment water. CONCLUSIONS: Microorganisms colonize dental units and contaminate dental treatment water. While documented instances of related illness are few, water that does not meet potable-water standards is inappropriate for use in dentistry. CLINICAL IMPLICATIONS: Exposure to water containing high numbers of bacteria violates basic principles of clinical infection control. Dentists should consider available options for improving the quality of water used in dental treatment. PMID- 11042983 TI - Dental safety needles' effectiveness: results of a one-year evaluation. AB - BACKGROUND: Some government agencies and state legislatures recently have passed regulations mandating the use of safety-enhanced devices, including dental anesthetic safety needles. Little information exists, however, on the efficacy and utility of these types of needles currently on the market. METHODS: The authors evaluated four types of dental safety needles and syringes for clinical acceptability. Two of these devices were deemed unacceptable owing to inherent features identified during the bench test. The remaining two devices were clinically evaluated using an 11-statement survey. Senior dental students completed the survey at one, two, four, five, six and eight weeks from introduction of the devices to a dental school clinic. Junior dental students joined the senior students using one of the devices for the last six months of the evaluation and joined the senior students in completion of a final survey at 52 weeks. RESULTS: The survey results indicated increasing user dissatisfaction with nine of the safety device features evaluated over the 52 weeks. At eight weeks, use of one of the two devices was discontinued owing to poor clinical performance. A review of the blood exposure incident reports that routinely are collected following an exposure incident revealed a small increase in exposures involving anesthetic needles. The sample size was too small to determine statistical significance of the change in injury rate, but it did show that needlesticks continue to occur in spite of the use of safety devices. CONCLUSIONS: None of the safety devices tested successfully passed the clinical evaluation. Continued evaluation is necessary to ensure that effective safety devices are available to dental practitioners. CLINICAL IMPLICATIONS: Evaluators had significant concerns about the usability of dental safety needles and their ability to adapt to using them effectively. Results of a review and bench tests indicate that the devices tested are no safer than traditional anesthetic needles. PMID- 11042984 TI - Anxiety and pain measures in dentistry: a guide to their quality and application. AB - BACKGROUND: The authors review measures of anxiety and pain used in recent dental studies. In particular, the study identifies the reliability, validity and usefulness of the measures. TYPE OF STUDIES REVIEWED: Three computerized databases of published scientific literature were searched over a 10-year period. Only studies that included measures of anxiety or pain were included. RESULTS: Information on the reliability and validity of 15 measures of dental care anxiety and three measures of pain and pain-related behaviors is provided. Reliability and validity data for most measures are good. Corah's Dental Anxiety Scale is the most widely used measure of anxiety, although it may not be as sensitive as other measures. The McGill Pain Questionnaire is the measure of choice for the assessment of pain. CLINICAL IMPLICATIONS: The authors have summarized properties of the scales for clinicians and researchers planning to use measures of anxiety, measures of pain, or both. PMID- 11042985 TI - Restoration fractures, cusp fractures and root fragments in a diverse sample of adults: 24-month incidence. AB - BACKGROUND: There are few reports in the literature regarding estimates of the occurrence of tooth fractures. Most studies have assessed tooth fractures in people seeking dental care, which may underestimate the incidence of the problem. METHODS: This study sought to estimate the incidence and prevalence of cusp and restoration fractures, as well as root fragments in participants in the Florida Dental Care Study, a diverse representative sample of community-dwelling residents of four north Florida counties made up of people who seek dental care regularly and those who do not. Participants received a dental examination and an in person interview at baseline and at a 24-month follow-up session. RESULTS: At least 20 percent of the participants were diagnosed as having bulk restoration fractures, cusp fractures or root fragments at the baseline visit. At the 24 month follow-up session, 26 percent of the participants had at least one new occurrence of these problems. Of those subjects presenting with tooth fracture, 25 percent had multiple teeth affected. African-Americans and people who sought care on a problem-oriented basis experienced twice the rate of cusp fracture and a higher rate of root fragments as did those who sought care on a regular basis. These data represent consecutive prevalence estimates rather than the true incidence, in that fractures that occurred after baseline may have been treated in the intervening 24 months. Thus, these data represent "lower-bound" incidence estimates. CONCLUSIONS: These results suggest that restoration fractures, cusp fractures and root fragments are a significant dental health problem, and that selected segments of the population are at greater risk of developing these problems. CLINICAL IMPLICATIONS: These data are useful for dentists in understanding the magnitude of the problem and the potential progression of fractures and root fragments. PMID- 11042986 TI - Enhanced dental cutting through chemomechanical effects. AB - BACKGROUND: Previous studies have shown that certain surface-active agents- compounds that reduce interfacial tension--in a dental handpiece's irrigation water can enhance cutting rates, or CRs. This study evaluated these effects under test conditions simulating dental practice. METHODS: The authors used a self contained cutting system with a digitally controlled handpiece speed, torque and water flow rate to cut machinable glass ceramic (Macor, Corning Inc.) with medium grit diamond burs and cross-cut fissure carbide burs under a load of 147.5 grams and 22 milliliters per minute coolant flow rate using water with mouthwash (Scope, Procter & Gamble) additions. They used six burs for each irrigant mixture to make three 5-millimeter edge cuts through 13 mm of Macor; CRs were quantified as the time necessary to transect the Macor cutting substrate. RESULTS: Additions of small amounts of mouthwash to the coolant water accelerated the CR for both carbide and diamond burs. The CRs for carbide burs in millimeters per second were distilled water, 0.21; 1:2.5 mouthwash:distilled water mixture, 0.12; 1:5 mixture, 0.64; and 1:10 mixture, 0.66. The CR differences for the 1:5 and 1:10 mixtures were significant (P < .001). The CRs for diamond burs in millimeters per second were distilled water, 0.09; 1:1 mouthwash:distilled water mixture, 0.13; 1:2.5 mixture, 0.16; 1:5 mixture, 0.21; and 1:10 mixture, 0.18. When it came to the diamond burs, the CR differences between water and the mouthwash:distilled water mixtures were significant (P < .001). The authors found that the mouthwash additions ensured higher CRs compared with those for water alone over the entire cutting regimen; that is, while the CRs for both carbide and diamond burs dropped with prolonged cutting with water irrigation, the addition of mouthwash resulted in the burs' cutting faster and for longer than with water alone. CONCLUSION: Adding small amounts of mouthwash to the coolant water significantly enhanced cutting by diamond and carbide burs and maintained higher CRs with prolonged cutting. CLINICAL IMPLICATIONS: Making low additions of mouthwash (1:5 and 1:10 mouthwash:distilled water mixtures) to the handpiece irrigant system can lead to two- to threefold increase in the dental diamond and carbide bur cutting rate compared with that for water alone. PMID- 11042987 TI - Enhancing the retention of prefabricated metal posts and resin cores. PMID- 11042988 TI - Composite restoration wear analysis: conventional methods vs. three-dimensional laser digitizer. AB - The purpose of this study was to compare three-dimensional laser digitizing with subjective evaluations of wear in posterior resin-based composite restorations during a 24-month period. BACKGROUND: The authors describe an indirect method of analyzing wear in resin-based composite restorations. A computer-driven laser was used to scan stone casts of restorations and create three-dimensional computerized surface models of the teeth. A standard computer algorithm was used to superimpose the follow-up model images over the baseline model images and to calculate the amount of wear within 10 micrometers. METHODS: Patients were selected whose treatment plans required a Class II restoration. A total of 100 restorations were inserted at baseline and evaluated at six months, 12 months and 24 months. Polyether quadrant impressions were taken and casts were made. Twenty one casts were selected for the three-dimensional laser digitizing technique. The same casts were evaluated via subjective evaluations using sets of 18 calibrated standard models, following the method developed by Leinfelder. RESULTS: Statistical analysis using the Wilcoxon test revealed significant differences (P < .01) between subjective wear evaluation and the three-dimensional laser digitizing wear analysis. CONCLUSION: The normalized three-dimensional laser digitizing technique is significantly more effective than subjective evaluations in establishing restoration wear rates. PMID- 11042989 TI - A clinical evaluation of 10 percent vs. 15 percent carbamide peroxide tooth whitening agents. AB - BACKGROUND: Agents with carbamide peroxide, or CP, in various concentrations are widely prescribed for at-home tooth whitening. It is not clear, however, if the more concentrated gels will whitening teeth to a greater extent, as no controlled clinical trials have been reported. The authors conducted a double-blind study of human subjects to evaluate whether a 15 percent CP tooth-whitening system was more effective than a 10 percent CP system, and to determine if tooth sensitivity increased with use of the higher concentration. METHODS: The authors recruited 57 subjects with maxillary anterior teeth of shade A3 or darker (as gauged against a value-oriented shade guide). The subjects were 18 to 65 years of age and in good general and dental health. After matching the subjects by sex and age, the authors randomly assigned them to either a control group, which used a 10 percent CP whitening agent, or an experimental group, which used a 15 percent CP agent. RESULTS: The results indicated that there was no significant difference in shade change between the groups after one week of treatment (t = 1.455, P = .05), but there was a significant difference at the end of the treatment period (t = 2.303, P < .05), as well as two weeks after treatment concluded (t = 2.248, P < .05). There was no significant difference in sensitivity (t = 1.399, P > .05). CONCLUSIONS: There was a significant difference in color change between the 10 percent CP and 15 percent CP groups at the end of the study period. There was no significant difference in level of tooth sensitivity between the two groups, and the incidence was equal; there was, however, a significant difference in variability of tooth sensitivity between the two groups. CLINICAL IMPLICATIONS: If performed under the careful guidance of a dentist, at-home whitening is an effective treatment, regardless of whether 10 percent CP or 15 percent CP is used. There may be added color change and varying sensitivity with the use of 15 percent CP. PMID- 11042991 TI - Elective vs. mandatory dentistry. AB - A major portion of the dental treatment provided today for the American consumer is elective. Dental patients are confused when they receive diverse treatment plans from different dentists. If treatment plans for oral therapy are divided into mandatory and elective categories, dental patients experience far less confusion. In this article, I have discussed methods of implementing this form of treatment planning. Dividing treatment plans into categories of mandatory and elective care has advantages for dental patients, the dental team and the profession as a whole. PMID- 11042990 TI - Dentists' participation in capitation and preferred provider organization dental plans. AB - BACKGROUND: In 1998, the American Dental Association Survey Center conducted a telephone and mail survey of U.S dentists in private practice in an effort to determine the extent of dentists' participation in capitation and preferred provider organization, or PPO, dental plans and the characteristics of dentists who participate in those plans. METHODS: An initial phone screening survey was conducted with a random sample of 11,550 dentists in private practice. Dentists who indicated that they participated in capitation or PPO dental plans received a follow-up mail survey asking specific questions concerning these two types of dental plans. RESULTS: Almost one-half of responding dentists indicated that they participated in either capitation or PPO dental plans. However, far more dentists reported participating in PPO dental plans than in capitation dental plans. The majority of participating dentists' patients were reported to be fee-for-service patients. CONCLUSIONS: Dentists' participation in PPO dental plans generally increased from that indicated in previous surveys, though participation in capitation plans declined. There was some regional and demographic variation in participation in these dental plans, but such differences were not large. Pricing and concerns about quality of care continue to be the primary concerns of nonparticipating dentists. PRACTICE IMPLICATIONS: Dentists reporting participation in PPO dental plans are becoming more common, but such plans still do not cover the majority of participating dentists' patients. A large percentage of nonparticipating dentists cite pricing and concerns about quality care as reasons for not joining these plans. PMID- 11042992 TI - MSOs revisited. PMID- 11042993 TI - Providing building services for healthy occupants? PMID- 11042994 TI - Diabetes mellitus type 2, obesity and weight loss. PMID- 11042995 TI - Health risks of air travel. PMID- 11042996 TI - Managing clinical waste--a continuing challenge for hospitals. PMID- 11042997 TI - Update on seasonal affective disorder (SAD). PMID- 11042998 TI - Oral cancer. PMID- 11042999 TI - Health and cancer prevention: implications of the knowledge and beliefs of 15-16 year old school pupils in South Africa. AB - The objective was to collect information from African and white pupils aged 15-16 years on their knowledge of and attitudes towards cancer, and their understanding of health-related behaviours in relation to their future experience. Questionnaires were completed by a series of 338 African and 378 white pupils in suburban secondary schools. African and white pupils knew most about lung cancer, and had some knowledge of skin and breast cancers. Smoking was seen by most as the chief cause of the disease. Half of both groups perceived a high intake of vegetables and fruit as protective. Television and printed media were the most important sources of information. The children in both groups were more anxious about unemployment and violence, and also, among African children, about AIDS, than about future ill-health. Thus, health was not perceived by many as the most important goal in life. Discussion indicates that white adolescents and also white adults, with more advanced understanding of cancer causation, make very limited use of their knowledge, dietary and non-dietary, to avoid the disease. In brief, level of cancer knowledge has very limited implications. Although young Africans in the course of their transition are very unlikely to seek to diminish their risk of cancer, there must be no lessening of urgent warnings and advice through the media on the avoidance of the disease. PMID- 11043000 TI - Against women: are we looking after our general practitioners? GPs' views of the 1990 part-time contract. AB - A postal survey was conducted looking at the roles and experiences of General Practitioners on part-time contracts. This involved their perception of the attitudes of their colleagues and patients to their part-time status, and the consequences of these for their professional development. Of the 130 General Practitioners with part-time contracts in the one Regional Health Authority that was being surveyed 77.7% responded; 74.3% of the respondents were women. Of the women General Practitioners who responded to this particular question, the predominant age-bands were 31-40 years [41.4% (41)] and 41-50 [19.2% (19)], whilst male General Practitioners were more evenly spread across the age bands. The results showed that the majority of General Practitioners took up part-time contracts to enable them to look after their dependants, though a sizeable minority wished for free time or to relieve stress from a full-time contract. Forty percent said that they felt excluded from decision making about continuity of patient care and practice policy. Just under forty percent also stated that their workload was excessive in comparison with their full-time counterparts. Further, many expressed the opinion that they were financially penalised. The cost effective correlation between the increased availability of General Practitioners (particularly women) for patient care, and the costly medical education and training of such General Practitioners not being 'wasted' for several years was also noted together for the need for ongoing (or continual) medical education and training. The findings of this survey suggest there are many unresolved issues involved in satisfactory part-time contracting arrangements for General Practitioners. This particularly affects women General Practitioners. Whilst the RCP policy statement addresses education and training for general practice, the question of not losing out in relation to training opportunities and promotion is not addressed. The unresolved effects of the intra professional differences in opportunities may affect the inter-professional functioning of the primary health care team and ultimately continuity and quality of care for patients. PMID- 11043001 TI - The impact of water supplies and sanitation on growth in Chinese children. AB - In many developing countries, improvement in water supplies has not been supplemented by improvement in sanitation facilities. Moreover, health education is rarely included in environmental hygiene programmes. Community health workers need to know if water supplies and sanitation have independent or complementary effects on health. This study analysed the weight data of 1,045 Chinese children aged 60 months or below. Regression models with interaction terms were tested against a model with main effects only. There was no evidence of interaction between water supplies and sanitation measures. The results show that water supplies and toilet facilities had independent associations with growth. Improved water source (P = 0.01) and flush-toilet (P = 0.06) were found positively associated with the children's weight. Presence of excreta in the home had a negative, but not statistically significant, association with weight. PMID- 11043002 TI - Self-reported smoking habits, biochemical markers, and nicotine dependence in a sample of the Danish population. AB - This study reports the smoking habits in a Danish population, evaluates plasma cotinine and thiocyanate levels in the detection of 'slips' (subjects participating in smoking cessation trials who begin to smoke a few cigarettes per week) and provides distribution scores on questionnaire measures of nicotine dependence. A total of 599 subjects with a mean age of 41 years participated in the study. Of these 46% were current smokers with no difference in the proportion of males(46%) and females(45%) and with a mean cigarette consumption of 12.7 daily. Plasma samples were analyzed for cotinine and thiocyanate, and the smokers completed two questionnaires to measure nicotine dependence: the Fagerstrom Tolerance Questionnaire and the modified Horn-Russell scale. The mean plasma cotinine was 207 micrograms/l for smokers, 14.4 micrograms/l for occasional smokers and 8.0 micrograms/l for non-smokers (ex-smokers and never-smokers). For plasma thiocyanate the levels were 130 mg/l, 54.8 mg/l, and 54.3 mg/l, respectively. The mean Horn-Russell score was 7.4 and the mean Fagerstrom score was 5.9. The two tests correlated with a t-value of 0.61 (p < 0.001) and the scores in both tests increased with increasing cigarette consumption. In conclusion, 75% of the smokers consumed 10 or more cigarettes per day and males smoked more cigarettes than females. It was impossible to distinguish occasional smokers (slips) from non-smokers using plasma cotinine or thiocyanate levels. We suggest that studies are needed to evaluate if light smokers benefit from nicotine replacement therapy because they achieve plasma cotinine levels which are similar to those seen when using patches for nicotine replacement therapy. PMID- 11043003 TI - Immunisation coverage at a well-baby clinic in the United Arab Emirates. AB - The United Arab Emirates Survey of Immunisation Coverage by UNICEF in 1990 showed that the coverage for UAE infants up to one year of age was 60%. The objective of this study was to determine factors associated with attendance and immunisation uptake at a well-baby clinic. A birth cohort of 665 infants were selected and their records examined to determine the factors such as gender, distance of housing area from clinic and access to other primary health care facilities. Only 22% attended all the scheduled visits and 54.3% of the sample completed the immunisation program within the correct time. The distance the infant lived from the clinic was the only factor significantly affecting immunisation uptake. The identification and targeting of groups at high risk of delayed completion of the immunisation program is a necessary risk factor in improving future surveillance. PMID- 11043004 TI - Hypohidrosis in children. AB - Hypohidrosis has many causes. The condition may be generalised or segmental, acute or progressive and central or peripheral in origin. Most of the causes can be diagnosed from the history and physical examination. Laboratory investigations are usually not necessary. Treatment should be directed at the underlying cause whenever possible. PMID- 11043005 TI - Ojirami-Ugbo: the village where the god of cleanliness is worshipped. AB - The beliefs and practices of the people of Ojirami-Ugbo, who worship Atta, the god of cleanliness, have been reported. A laboratory survey showed significant medical benefits in those health measures adopted by the people. PMID- 11043006 TI - A methodology for setting practice criteria in healthcare. AB - Practice criteria are an important part of health care and have taken a new prominence in the trend to address quality-of-care issues. Once an organisation makes a commitment to addressing its quality-of-care, it must define 'quality' in operational terms. Practice criteria do just that. The organisation ensures consistent, high-quality services through the correct application of practice criteria. This paper outlines a methodology that has been used in at least three countries to date. Early indications are that it is useful for helping an organisation begin its quality improvement 'journey'. PMID- 11043007 TI - Food hygiene for students whose preferred language is not English. PMID- 11043008 TI - Lessons to be learned: a case study approach: severe hyponatraemia induced by primary hypothyroidism and associated with possible increased hepatic sensitivity to thyroxine replacement. AB - The case is presented of a 74 year-old woman who was admitted with severe hypo osmolar hyponatraemia associated with inappropriately raised urinary osmolality, and who was subsequently discovered to have primary hypothyroidism. A normal serum sodium concentration was restored by means of judicious fluid restriction and thyroid hormone replacement. Low dose thyroxine therapy led to rapid but modest increases in the serum activities of alanine aminotransferase (ALT) and alkaline phosphatase (ALP); both returned to normal over a period of three weeks. These sub-clinical enzyme changes may indicate tissue 'hyperthyroidism'; and in this case, the fact that they occurred acutely at only low doses of thyroxine possibly suggests an increased hepatic sensitivity to the hormone. PMID- 11043009 TI - Historical perspectives on health: assessment of national progress in public health in England, 1832-1911. AB - The health of society is the sum of recent public policy and the heritage of public health and welfare. The degree of improvement in the health of young males in the nineteenth century may be gauged by scrutinizing changes in the ratio of weight to height2 (W/H2). The two variables summate the developmental history of individuals, and so are an index of biosocial circumstances for a set of people. Such data are uncommon, and this essay formulates the ratio for the years 1832 to 1911. The conclusion is that, according to the W/H2 criterion, innovations in public policy produced improvement in the health of young males. PMID- 11043010 TI - Re: Respiratory symptoms and lung function in garage workers and taxi drivers, December 1998 (J Roy Soc Health; 118 (6); 346-353). PMID- 11043011 TI - Re: The challenge of hospital nutrition, current topics, March 1999 (J Roy Soc Health; 119 (1); 9-10). PMID- 11043012 TI - Prevention of fire ant anaphylaxis in South Carolina: improving patient referrals, 1989-1999. PMID- 11043013 TI - Seizures in children: a guide for pediatricians and general practitioners. AB - Seizures are very common in children and adolescents. Physicians dealing with young seizure patients should develop a well-organized and step-wise approach to the diagnosis, bearing in mind that other paroxysmal disorders can closely mimic epileptic seizures. Treating physician should become familiar with a few of the many anticonvulsants available and use them judiciously in selected cases. PMID- 11043014 TI - New legislation covers reimbursement and diabetes care guidelines. PMID- 11043015 TI - An outpatient technique for continuous subcutaneous insulin infusion in adolescents and young adults with IDDM. PMID- 11043016 TI - Paradigm shifts in health care. PMID- 11043017 TI - The efficacy and safety of cefaclor in respiratory infections amongst Pakistani children. AB - PURPOSE: To evaluate the efficacy and safety of Cefaclor in respiratory tract infections amongst Pakistani children. PATIENTS AND METHODS: Multicenter, open label and non-comparative study was done to evaluate the response in terms of symptoms (In vivo) and bacterial cultures (In Vitro) to Cefaclor amongst children with respiratory tract infection between the ages 2 months to 12 years. Each patient was asked to visit the doctor on three occasions i.e., Day 0 (Initial evaluation prior to commencement of study), Day 4 (During therapy assessment and confirmation of compliance) and Day 10 (End of therapy assessment and compliance evaluation). Representative swab specimens (Throat swabs, Ear swabs or Sputum) were collected from the infected site on day 0 and day 10 for culture and sensitivity. Patients were also assessed by the evaluators on each visit in terms of clinical symptomatic response and information collected was documented on a prescribed data base form. RESULTS: A total of 160 patients were enrolled in the study, of whom 15 were lost to follow-up between the first and second visit and a further 38 were lost by the 3rd visit. Thus 107 patients completed the study as per protocol. Otitis media and Upper respiratory tract infection were the predominant ailments amongst the cases enrolled. One or more bacteria were isolated in 75 (46%) instances, the maximum number of isolates being from ear swabs of Otitis media patients. Beta haemolytic Streptococcus (group A,C,F,G) seen in 18 cases was the most common pathogen reported followed by Staphylococcus aureus, H. influenzae and Streptococcus pneumoniae in 13,12 and 11 cases respectively. Sensitivity of Cefaclor for bacteria commonly seen in the respiratory tract was greater than 90% in most of the cases. Evaluation of the 42 culture proven cases for patients who completed the study showed that Cefaclor had a 93% efficacy for indicated bacteria and 54% for non-indicated bacteria. In Vivo analysis of Cefaclor (i.e. on the basis of symptomatic response) showed that 96% cases had a symptomatic response by the second visit, which improved to 97% by the third visit. Only 15 non-serious adverse events were observed in 160 patients, none of the cases necessitated discontinuation of drug. Mild gastrointestinal symptom was the most common adverse event reported. CONCLUSION: Cefaclor was found to be a safe and efficacious drug in the treatment of bacterial respiratory tract infections amongst Pakistani children. PMID- 11043018 TI - Genetic diversity of beta-thalassemia mutations in Pakistani population. AB - BACKGROUND: beta-thalassemia is one of the most common inherited single gene disorder in Pakistan. It is characterized by reduced or absent beta-globin gene expression resulting in abnormal maturation and survival of red blood cells. Due to high prevalence of this disease in the local population, it has become important for the health care providers to encourage people to utilize laboratory facilities for carrier and prenatal genetic testing. OBJECTIVE: To study the frequency of beta-thalassemia mutations in Pakistani population. SETTING: A tertiary care teaching hospital. METHODS: Blood samples of 72 couples and chorionic villus (CV) biopsy specimen collected at the Aga Khan University Hospital, Karachi were tested by Amplified Refractory Mutation Systems (ARMS) for the 12 most common mutations in the beta-globin gene. RESULTS: Out of 72 chorionic villus biopsy specimen analyzed, 17 (23%) had mutations in both alleles of the beta-globin gene. Homozygosity was identified in 6 CV samples, whereas 11 CV specimens were diagnosed as double heterozygous. Almost 60% of the CV biopsies showed mutations in one allele and were diagnosed as carriers. IVSI-5 (G-C) was the most common mutation identified in this study. It was found in 53% of the subjects and was represented equally in all the ethnic groups except Pathans. Several regional and ethnic differences were observed in the distribution of common mutations, for example in Pathan families Fr 8-9 (+G) mutation was most prevalent. In addition, variation in the distribution of mutations was also observed between the Northern and the Southern regions. CONCLUSION: This study indicates that in Pakistan, the five most common mutations are IVS1-5 (G-C), IVS1 1 (G-T), Fr 41-42 (-TTCT) Fr 8-9 (+G) and deletion 619 bp. An important factor contributing to high incidence of thalassemia is the unawareness among people about the available diagnostic facilities for the prenatal diagnosis in Pakistan. Strict implementation of collective measures including carrier identification, genetic counseling and prenatal diagnosis are required for preventing beta thalassemia in Pakistan. PMID- 11043019 TI - Effectiveness of health education in promoting the use of iodized salt in Lotkoh, tehsil Chitral, Pakistan. AB - INTRODUCTION: We evaluated the effect of health education on the use of iodized salt in a remote region. METHODS: We randomly selected 31 villages in teh Lotkoh tehsil of district Chitral in the North West Frontier Province of Pakistan. We then randomly selected 7 households from each village and inteviewed the eldest women of the family. We also tested samples of salt for iodine concentration at the user's level. RESULTS: Eighty-five percent of families (184/217) used iodized salt exclusively. Among the samples population, the Aga Khan Health Services (AKHS) informed 67% about the importance of iodized salt. Shopkeepers and neighbors informed 25%. People informed by AKHS were more likely to know the volatile nature of iodine (76% vs 55%, p < 0.001) and the advantages of iodized salt (91% vs. 75%, p = 0.001) than persons informed by other sources. People who could name any single advantage of iodized salt were more likely to use iodized salt (97%) compared to those who could not name any advantages (62%) (p < 0.001) Iodine concentration in 78% (141/183) samples was acceptable (> or = 15 ppm). One specific brand of salt consistently had sufficient iodine concentration (91%) compared to all others (47%) (p < 0.001). CONCLUSION: Health education has been effective in promoting the use of iodized salt in these isolated rural communities. A joint effort by the government, local NGOs and the community can substitute the role of mass media in such areas. Regular evaluation of iodized salt brands should be considered. PMID- 11043020 TI - Adult mortality in slums of Karachi, Pakistan. AB - OBJECTIVE: Cause-specific death rates are rarely available to guide health interventions for adults in South Asia. We report mortality patterns among Karachi's urban poor. METHODS: We conducted verbal autopsies for adult deaths under active surveillance during 1990-1993 in five urban slums of Karachi. Two physicians assigned underlying cause of death by consensus. Analysis included cause- and category-specific rates, 45Q15s and comparison with 1991 Japanese national statistics. RESULTS: All 345 adult deaths (15-59 years) in the 5 slums (total population 45,389) were included. Male mortality exceeded female (4.4 vs 3.3/1000, p = .02). Noncommunicable diseases claimed 59% of deaths, communicable and reproductive 27% and injuries, 15%. The leading identified death rates (/100,000) among women were: circulatory disorders (66), maternal causes (33), tuberculosis (30), and burns (23); and among men they were: circulatory disorders (124) tuberculosis (30) and road traffic accidents (30). Overall Karachi adult mortality was 3.7 times Japanese rate. Compared to Japan, adults in Karachi had one to two orders of magnitude excess mortality due to maternal causes, tuberculosis and burns. Circulatory disorders and tuberculosis accounted for 47% of excess male mortality; these plus maternal causes and burns accounted for 55% of excess female mortality. CONCLUSION: These mortality levels and patterns compel interventions and research for poor urban adults beyond maternal health. Women's health would equally benefit from tuberculosis control or burn prevention. Men need safer travel. Both need improved cardiovascular health. PMID- 11043021 TI - The minds of mothers: maternal mental health in an urban squatter settlement of Karachi. AB - BACKGROUND: Community-based information on maternal mental health in developing countries is meager and nearly non-existent in Pakistan. OBJECTIVE: To determine the proportion of probable cases of women with mental disorders and examine the associated conditions and risk factors which contribute to maternal mental ill health. METHODS: With convenient sampling 260 mothers in an urban squatter settlement of Karachi were interviewed. The tools consisted of a household questionnaire collecting information on basic demographic and other characteristics and the Aga Khan University Anxiety and Depression Scale (AKUADS), an instrument to assess psychiatric morbidity. RESULTS: The proportion of probable cases of mental disorder was 28.8% (n = 75). Reviewing the gradient of responses the most frequently expressed psychiatric symptoms were "being worried" and "crying". Amongst somatic complaints the most frequently reported was headache. Study also suggests that women in the older age group (OR 2.30, CI 1.27-4.19, p = 0.0031) and those with longer duration of marriage (OR 1.80, CI 1.01-3.22, p = 0.032) are more likely to be mentally distressed. Arguments with husband (OR 5.0, CI 2.19-11.52, p = 0.00001) or in-laws (OR 2.43, CI 1.22-4.85, p = 0.0059), husband's unemployment (OR 4.1, CI 1.27-13.6, p = 0.0058), not having permanent source of income and lack of autonomy in making decisions significantly contributed towards mental illness. CONCLUSION: Approximately 1 out of 4 women suffer from mental illness. This is alarmingly high. Besides counseling in cases of matrimonial disharmony, community-based interventions should aim to improve the socioeconomic status of households. PMID- 11043022 TI - Domestic violence against women--perspective from Pakistan. AB - OBJECTIVE: Domestic violence is an important public health concern. This study assessed the prevalence and type of domestic violence perpetrated by men on their wives. SETTING: The study was conducted in a public sector hospital among men accompanying patients. METHODS: A cross-sectional survey based on a sample of convenience was conducted by the author, using a pre-tested questionnaire with mostly close-ended questions. RESULTS: All the respondents admitted to ever shouting or yelling at their wives, including while she was pregnant. Twenty three (32.8%) respondents admitted to ever having slapped their wives and 54 (77.1%) admitted to ever engaging in a non-consensual sex with their wives. CONCLUSION: Population based studies are required to assess the type, frequency, risk factors and sequel of wife abuse, so as to establish practice guidelines. PMID- 11043023 TI - Surgical management of penile cancer--report of two cases. PMID- 11043024 TI - Low serum alpha 1 antitrypsin in duodenal ulcer--a family study. PMID- 11043025 TI - Staphylococcus aureus meningitis in a post splenectomy patient. PMID- 11043026 TI - Intravenous immunoglobin G therapy in Evans syndrome. PMID- 11043027 TI - Place research problem in theory--a theory research symbiosis. AB - Theory gives a specific perspective to research by providing systematic guidance for the identification and investigation of the logical relationship in the phenomenon of interest. To critically appraise the theoretical bases of a research, it is important first to familiarize with the language of theory, which mainly comprises of concepts and propositions. Furthermore, models are developed to act as a springboard for theory generation and theory practice. To organise the research findings in order to develop a body of knowledge, the connection between theory and research should be explicit in the study. PMID- 11043028 TI - U.S. Supreme Court validates HMO financial incentives and further undermines professionalism in the practice of medicine. PMID- 11043029 TI - Role of the physician in the home health prospective payment system. PMID- 11043030 TI - Cerebral air emboli: an uncommon complication resulting from a common procedure. PMID- 11043031 TI - [Shoulder tendon disorders]. PMID- 11043032 TI - [General notions on epicondylalgias and epitrochlealgias]. PMID- 11043034 TI - [Epicondylitis in 35 amateur athletes]. PMID- 11043033 TI - [Epicondylitis in athletes]. PMID- 11043036 TI - [Tendon disorders of the wrist and hand: principal localizations]. PMID- 11043035 TI - [Wrist tendon disorders. General notions]. PMID- 11043038 TI - [Achilles tendon disorders]. PMID- 11043037 TI - [Tendon disorders of the knee]. PMID- 11043039 TI - [Ankle tendon disorders outside of the Achilles tendon]. PMID- 11043040 TI - [Foot orthoses in foot and ankle tendon disorders]. PMID- 11043041 TI - [General notions on Achilles tendon rupture]. PMID- 11043043 TI - [Hospital hygiene--in the middle of the storm]. PMID- 11043042 TI - [Achilles tendon rupture in athletes. Ten cases]. PMID- 11043044 TI - [Death among drug addicts]. PMID- 11043045 TI - [Better understanding of the biology of cancer cells]. AB - Most forms of cancer arise through a Darwinian evolutionary process. The natural selection that ultimately leads to cancer takes place in somatic tissues although it may be triggered by inherited mutations in a small but significant minority. It favors the growth of clones and subclones that are less and less responsive to normal intra- and extracellular growth control mechanisms. The development of molecular biology has led to the identification of many genes that participate in this somatic evolution. They belong to the following groups: Oncogenes, constitutively activated by structural and/or regulatory changes that drive the cell to continuous proliferation; Tumor suppressor genes, that can inhibit the illegitimately activated cell cycle. They contribute to tumor development by loss mutations or permanent down-regulation, e.g. by methylation; Apoptosis inhibitory genes that can contribute to tumor development by raising the apoptotic threshold, and apoptosis promoting genes that can favor the growth of apoptosis prone tumor cells by their loss or inactivation; DNA repair genes whose inactivation can counteract the normal elimination of cells that carry potentially cancer promoting mutations. Inherited mutations in DNA repair genes can lead to familial cancer syndromes. Immortalizing genes that counteract cellular senescence; Angiogenesis promoting genes whose products may stimulate the vascular supply of tumors; Genes whose structural or functional changes may facilitate the escape of tumor cells from immune rejection; The multi-step development of individual tumors can encompass changes in most or all of these genes. They occur independently of each other and without any fixed order or timing. Tumor emancipation from growth control can therefore proceed along various pathways. It follows that each tumor must be regarded as a biologically unique individual. PMID- 11043046 TI - [Fatal intoxications among drug addicts in Denmark in 1997]. AB - INTRODUCTION: The purpose of this study was to investigate fatal poisonings among drug addicts in 1997 and to compare the results to similar investigations from 1985 and 1991. MATERIAL AND METHODS: All fatal intoxications among drug addicts in Denmark in 1997, investigated at the three Institutes of Forensic Medicine in Denmark. RESULTS: The number of fatal intoxications increased by 32% from 1991 to 1997, mainly outside the metropolitan area, The average age increased from 32 to 36 years. The proportion of heroin/morphine intoxications increased from 57% to 71%. The most commonly used drugs were as in 1991 heroin/morphine, diazepam and methadone. The frequency of cocaine increased from one positive case in 1991 to 14% positive cases in 1997. DISCUSSION: This study showed an increasing number of fatal intoxications and changes in drug abuse pattern and place of death since 1991. PMID- 11043047 TI - [Surgical mesh implantation in incisional hernia. A historical follow up study]. AB - AIM OF STUDY: To describe risk factors, recurrence rates, morbidity and satisfaction in patients operated for incisional hernia with implantation of a surgical mesh. MATERIAL AND METHODS: The records of 60 patients undergoing an incisional hernia repair using a surgical mesh in the period 1.1.1993-30.6.1998 were reviewed, and 49 living patients were examined. RESULTS: During a mean follow-up of 27 months, patients with preoperative risk factors had more complications and recurrences. Overall recurrence rate was 14%. Ninety-six percent were satisfied with the operation and 90% wanted the same operation in case of recurrence. CONCLUSION: Operation for incisional hernia using implantation of a surgical mesh is a safe and well tolerated procedure, care must be taken in patients with risk factors. PMID- 11043048 TI - [Metaphors and power. A qualitative study of metaphorical thinking among Danish cancer patients]. AB - The use of metaphors concerning cancer was investigated by interviewing nine Danish cancer patients, aged 30-85 years. Metaphors about the disease were widely used, e.g. personification metaphors. Most patients considered their cancer as a metaphor for a physical and psychological imbalance in the period before the disease. Some patients felt responsible for the disease. The metaphors of cancer patients should be considered when communicating with cancer patients. PMID- 11043049 TI - [Baker's cysts in children. A retrospective study at the hospitals in the county of Rebe from 1992 to 1998]. AB - Popliteal cysts or Baker's cysts in childhood is a rare disease usually found by the parents. The purpose of this study is to study the recurrence rate after primary surgical resection and conservative treatment of the cysts. We retrospectively found 20 children with Baker's cysts treated at Ribe Amts Hospitals during the period 1992-1998. Thirteen patients were treated conservatively and seven had a surgical resection of the cysts performed. Of the 13 conservatively treated patients the cysts was still present in six patients (46%). In seven patients the cysts had disappeared within nine months. In the surgically treated group the cysts recurred in three patients (43%). Surgical intervention for Baker's cysts in children should only be performed after thorough consideration, due to a high recurrence rate. PMID- 11043050 TI - [Hantavirus disease during pregnancy]. AB - Hantavirus infection is known to occur in mice and rats without symptoms. In contrast, an infection with Hantavirus in humans gives influenzalike symptoms and acute renal failure. The treatment is symptomatic, but treatment with dialysis can be necessary for a while. A case of Hantavirus infection in Denmark in a 30 week pregnant woman is reported. PMID- 11043052 TI - [Picture of the month. Colonic adenoma]. PMID- 11043051 TI - [Thoracic aortobifemoral bypass in chronic lower limb ischemia]. AB - In cases of critical limb ischemia caused by aorto-iliac arteriosclerosis the most common operation is aortobifemoral bypass. This operation has a high patency. We describe two cases in which a thoracic aortobifemoral bypass was chosen as an alternative bypass operation because of a hostile abdomen. Both operations were performed without complications and resulted in normally perfused extremities. PMID- 11043053 TI - [Transport receptors can signal, too]. PMID- 11043054 TI - [Amebiasis]. PMID- 11043055 TI - [NSAID and fluid retention. Is it possible to change to other NSAID?]. PMID- 11043056 TI - [Leflunomide for active rheumatoid arthritis]. AB - In 1999, leflunomide was introduced for the treatment of active rheumatoid arthritis. Leflunomide is a reversible inhibitor of "de novo" synthesis of pyrimidine, resulting in a restriction of lymphocyte proliferation. The pharmacodynamics are characterized by slow elimination. Leflunomide has shown an efficacy and a pattern of mild and serious side effects similar to methotrexate and sulphasalazine. Treatment is started with a loading dose of 100 mg orally for three days followed by 20 (10) mg daily. Regular haematological measurements, determination of serum chemistry and blood pressure should be performed every second week the first six months, subsequently every sixth week. If the treatment is to be terminated abruptly, a drug elimination procedure must be performed. Leflunomide will find its place in the treatment of rheumatoid arthritis. Leflunomide should follow an unsuccessful treatment attempt with methotrexate or sulphasalazine. PMID- 11043057 TI - [Rofecoxib, a new NSAID preparation with selective COX-2 inhibition]. AB - NSAIDs are effective by inhibition of the enzyme cyclooxygenase, which leads to formation of prostaglandins. At the beginning of the 90-ties it became evident, that cyclooxygenase existed in two isoforms (COX-1 and COX-2). COX-1 is found in a number of tissues. In these tissues prostaglandins have several physiological functions. COX-2 is an induceable enzyme involved in inflammation. A new NSAID, rofecoxib, has selective COX-2 inhibition defined as inhibition of COX-2, but not COX-1 in the therapeutic dose range. Trial results have shown pain relief analogous to common NSAIDs (ibuprofen and diclofenac) in patients with osteoarthritis of the hip or knee. The incidence of gastric ulcers was significantly reduced in the rofecoxib treated group compared to the groups treated with common NSAIDs. It is concluded that selective COX-2 inhibition is a promising new treatment. At present it seems appropriate to consider the drug in patients with good indication for an NSAID and simultaneous gastrointestinal risk factors. PMID- 11043058 TI - [Oral transmucosal fentanyl citrate (OTFC) in the treatment of breakthrough pain]. AB - Breakthrough pain in patients receiving opioids for pain relief is traditionally treated with opioids given orally. This, however, implies a long time to clinical effect and a long duration of action resulting in difficult titration. It is possible to give fentanyl orally for transmucosal absorption (OTFC) with a new formulation using a stick with a specially designed tablet that can be rubbed against the mouth mucosa. This gives a short time to effect and with a short duration of action. Clinical studies have shown that patient tolerance of OTFC is high and the analgesic effect is comparable to that of intravenous fentanyl, without the need for an i.v. line. Clinical indication will be breakthrough pain in patients receiving opioids for baseline pain medication. PMID- 11043059 TI - [Tremor--in cattle and human beings--when exposed to growth promoters]. PMID- 11043060 TI - Public health series take 3: infectious disease PMID- 11043061 TI - Too many treatments for Parkinson's disease: how should they be used? PMID- 11043062 TI - What is healthy about the "Healthy People 2000" document? PMID- 11043063 TI - Infectious diseases in Wisconsin: a public health perspective. PMID- 11043064 TI - Overuse of antibiotics leads to "super bugs". PMID- 11043065 TI - Hepatitis C: a hidden, growing problem. PMID- 11043066 TI - Old foes, new enemies appear on infectious disease front. PMID- 11043067 TI - Escherichia coli O157:H7 infections in Wisconsin, 1992-1999. AB - Escherichia coli O157:H7 infection became a reportable condition in Wisconsin on April 1, 2000; previously cases were voluntarily reported by physicians and laboratories. During 1992 through 1999, 1333 cases of E. coli O157:H7 infection occurred in Wisconsin residents and were reported to the Wisconsin Division of Public Health. During this interval, the highest age-specific mean annual incidence, 13.2 cases per 100,000 population, occurred in persons 3 to 5 years old. Only 28% of patients with reported cases identified bloody diarrhea among their signs and symptoms. Of reported cases, 17% (231/1333) were involved in the eight outbreaks investigated during this interval. Among case patient identifiable risk exposures, farm related (13.4%), recreational water related (8.1%), and unpasteurized milk/dairy product (7.0%) exposures were the most frequently noted. Relatively few infections involved raw/undercooked ground beef consumption (5.8%). Recent use of pulsed-field gel electrophoresis has facilitated linkage of sporadically reported cases into recognized outbreaks. E. coli O157:H7 infections frequently occur in Wisconsin; acquisition of these infections in a wide variety of settings poses important challenges to their prevention and control. PMID- 11043068 TI - HIV infection in Wisconsin: an overview of epidemiologic trends, 1983-1999. AB - A review of HIV case surveillance data shows that the number of persons reported with HIV infection in Wisconsin steadily increased during the 1980s, leveled in the early 1990s and since 1993 has tended to decline. Cases reported in 1999 represented a 44% decrease compared to the 1990-1993 average. The number of deaths among persons with HIV infection declined 64% from 1993 to 1999; as a result, the number of persons living with HIV infection nearly doubled during the 1990s. Comparing cases reported 1995-1999 with cases reported in the 1980s, a higher percentage was attributed to injection drug use and high-risk heterosexual contact. A higher percentage of HIV cases also occurred among females and racial and ethnic minorities. PMID- 11043069 TI - Wisconsin, July 1999 heat wave: an epidemiologic assessment. AB - OBJECTIVE: To assess the characteristics of heat-related deaths in Wisconsin during the summer of 1999. METHODS: Review of death certificates indicating heat as an underlying or contributing cause of death. RESULTS: Heat-related illness led to 21 deaths during the summer of 1999 in Wisconsin. The rate of death was highest in the elderly, particularly those aged 65-84 years (2.2/100,000). Heat was the underlying cause for 12 of the 21 deaths. Cardiovascular conditions were the underlying cause in 8 of the deaths, and a contributing cause for another 7. CONCLUSIONS: The elderly, persons taking psychotropic medications, and persons with chronic diseases, particularly cardiovascular conditions, are at increased risk of death from heat during heat waves. Prevention messages and weather advisories during heat emergencies must target these groups. Care givers and medical personnel must be on heightened awareness for the signs and symptoms of heat exhaustion and heat stroke during these periods. PMID- 11043070 TI - Impact of Haemophilus influenzae type b (Hib) conjugate vaccines on Haemophilus influenzae meningitis in Wisconsin. AB - The first effective Haemophilus influenzae type b (Hib) conjugate vaccines were approved for use in children and infants between 1987 and 1990. In 1993, the federal government began the Childhood Immunization Initiative (CII), a program to improve the rate of vaccination of children nationwide. Subsequently, the proportion of 19 to 35-month-old children who received three or more doses of Hib vaccine rapidly increased from 28% in 1992 to 90% by 1995, with a concurrent dramatic decline in the incidence of H. influenzae meningitis. We reviewed cases of H. influenzae meningitis reported to the Wisconsin Division of Public Health from 1981 to 1997. The mean annual incidence of H. influenzae meningitis declined 96% from 2.4 cases per 100,000 persons in the pre-vaccination period (1981-1986) to less than 0.1 case per 100,000 persons after Wisconsin had achieved 90% Hib vaccination coverage (1994-1997). H. influenzae meningitis occurrence declined dramatically among all age groups, including 96% among children aged less than 1 year old, 99% among 1-4 and 5-9 year olds, and 46% among persons 10 or more years old. Consistent with national trends, the majority of H. influenzae meningitis cases reported in 1997 was caused by non-type b strains of H. influenzae. PMID- 11043071 TI - HIV infection in Wisconsin: state and national resources for clinicians and patients. AB - The authors review state and national resources for clinicians and patients regarding HIV infection, high-lighting pertinent resources in the Wisconsin AIDS/HIV Program as well as select local and national resources. PMID- 11043072 TI - Antibiotic susceptibility of invasive Streptococcus pneumoniae in Wisconsin, 1999. AB - BACKGROUND: Streptococcus pneumoniae is a major cause of community acquired infections in the United States, and rates of antibiotic resistance have increased dramatically in the past decade. Statewide rates of pneumococcal resistance to penicillin and other antibiotics have not been previously reported in Wisconsin. To determine these rates, we assessed invasive pneumococcal isolates for reduced susceptibility to nine different antibiotics. METHODS: Pneumococcal isolates from blood, cerebrospinal fluid or other normally sterile body sites were submitted by 91% of laboratories that perform invasive bacterial cultures. Isolates were tested for susceptibility to penicillin, cefotaxime, ceftriaxone, levofloxacin, meropenem, erythromycin, vancomycin, sulfamethoxazole trimethoprim and chloramphenicol. RESULTS: There were 409 invasive pneumococcal isolates identified in 1999 among Wisconsin residents, including 385 (94%) isolates from blood. The mean patient age was 42.5 years (range, < 1 year to 96 years), and 213 (52%) were male. Of the pneumococcal isolates, 24% were not susceptible to penicillin, including 10% with high level resistance. Isolates with reduced penicillin susceptibility were also likely to have reduced susceptibility to other antibiotics. Patients with penicillin nonsusceptible (intermediate and fully resistant) pneumococcal isolates were significantly younger (mean, 37.0 years) than those with susceptible isolates (mean, 44.3 years) (p = .04). The proportion of patients with a penicillin nonsusceptible isolate varied by region, ranging from 12.8% in northeastern Wisconsin to 35.5% in northern Wisconsin. CONCLUSIONS: The proportion of invasive pneumococcal isolates with penicillin resistance in Wisconsin is similar to other regions of the United States. Inappropriate antibiotic use contributes to the emergence of resistant pneumococcal infections, and educational efforts are underway to promote judicious antibiotic use in Wisconsin. PMID- 11043073 TI - Proceedings from the 1999 annual meeting of the American College of Physicians American Society of Internal Medicine, Wisconsin Chapter. PMID- 11043074 TI - MCW building on its commitment to genetic research. PMID- 11043075 TI - Peer review records may be discoverable in federal court. PMID- 11043076 TI - Reflections on an emergency department visit. PMID- 11043077 TI - Wireless communication is all around. It's gone way beyond cellular phones. PMID- 11043078 TI - Choosing a hair-removal method. PMID- 11043080 TI - Sinusitis and its relationship to asthma. Can treating one airway disease ameliorate another? AB - In a large percentage of patients with upper airway disease, asthma and sinusitis occur concomitantly. As many as 88% of patients with asthma exhibit the symptoms of rhinitis, and half of rhinitic patients have asthma. In this article, Dr Muller explores the interrelationship of sinusitis and asthma, including characteristics, mechanisms of action, and diagnosis, and provides suggestions for optimal treatment of both. PMID- 11043079 TI - Sudden cardiac death in young athletes. Causes, athlete's heart, and screening guidelines. AB - Sudden cardiac death of a young competitive athlete is a rare but tragic event. Hypertrophic cardiomyopathy and coronary artery anomalies are the most frequent causes. Most cardiovascular abnormalities go unrecognized until the time of death owing to the lack of preceding signs or symptoms suggestive of disease. Physicians responsible for the care of athletes should be familiar with the various causes of sudden cardiac death, the physiologic adaptations seen in so called athlete's heart, and existing cardiovascular screening guidelines. The preparticipation evaluation, although it has limitations, is the major instrument readily available for prevention of sudden cardiac death. Effort should be made to follow established consensus guidelines. PMID- 11043081 TI - Managing eye disease in primary care. Part 1. How to screen for occult disease. AB - Proper eye screening is the first step in detection of occult eye disease in asymptomatic patients. Knowing which patients are at high risk and should be referred for a comprehensive eye examination is the key. In part 1 of this three part article, Dr Shields explains how to identify high-risk patients and offers practical pointers for performing visual acuity measurements and other necessary testing in children. A list of resources on vision care for both patients and physicians is also included. PMID- 11043082 TI - Managing eye disease in primary care. Part 2. How to recognize and treat common eye problems. AB - Many patients with symptomatic eye conditions present initially to their primary care physician. Comprehensive treatment of such conditions as dry eye syndrome, blepharitis, styes, chalazia, conjunctivitis, congenital nasolacrimal duct obstruction, superficial foreign bodies, corneal abrasion, and subconjunctival hemorrhage can often be provided in the primary care setting. Patients with persistent or severe conditions, particularly those involving foreign bodies and corneal abrasions due to high-velocity injuries, should be referred for immediate care by an ophthalmologist. PMID- 11043083 TI - Managing eye disease in primary care. Part 3. When to refer for ophthalmologic care. AB - Most severe eye diseases and injuries ultimately require intervention by an ophthalmologist. The urgency of referral depends on various factors, including level of vision loss, duration of symptoms, and presence of comorbid diseases. Of special importance are five acute eye problems in which emergency management by primary care physicians can be critical to visual outcome: high-velocity injuries, chemical injuries, acute angle-closure glaucoma, arteritic ischemic optic neuropathy, and central retinal artery occlusion. PMID- 11043084 TI - Optimal antidepressant dosing. Practical framework for selection, titration, and duration of therapy. AB - Appropriate antidepressant dosing and trial duration are crucial for successful treatment of depression. Before prescribing an antidepressant, primary care physicians should take into account each patient's history, responses to previous antidepressants, depressive symptoms, coexisting illnesses, and current prescriptions. Physicians must be able to help patients manage side effects and know when to discontinue treatment, switch antidepressants, or refer patients to a psychiatrist. PMID- 11043085 TI - New insights into genetic aspects of Alzheimer's disease. Does genetic information make a difference in clinical practice? AB - Genetic testing sometimes offers definitive information for patients who have a family history of early-onset Alzheimer's disease that occurs before age 50 in a Mendelian pattern. However, for patients who are already symptomatic, especially those with sufficient symptoms to warrant a clinical diagnosis of Alzheimer's disease, genetic testing may not contribute a great deal of information beyond that already available from the clinical and family history. For prediction of disease onset, genetic testing can sometimes give a clear picture of disease risk, but each patient must carefully weigh the risks and benefits of having that information. For early-onset Alzheimer's disease occurring beyond age 50 or without a clear Mendelian pattern, genetic testing is unlikely to be informative. In patients who have a family history of late-onset Alzheimer's disease, while APOE's contribution to increased risk is indisputable, its potential use as a genetic test is very limited. Testing may be helpful as an adjunct to clinical diagnosis but does not obviate the need for a full workup for treatable causes. Thus, the benefit of testing may be marginal. No consensus has been reached as to the value of genetic testing for early detection of late-onset disease, but APOE testing might become important in the future if it helps to define the need for intervention or to select an optimal intervention. There is a broad consensus that APOE testing lacks sufficient predictive value to be suitable for predictive testing in asymptomatic persons. PMID- 11043086 TI - Understanding essential tremor. Differential diagnosis and options for treatment. AB - Although essential tremor is common, its various presentations may be confused with other movement disorders, such as Parkinson's disease and dystonic tremor. In this article, Dr Evidente describes classification of tremor, the clinical features of essential tremor, and the differential diagnostic considerations. He also discusses the extensive list of medications used to treat the disorder and the surgical options for severe, drug-resistant cases. PMID- 11043087 TI - Strategies for controlling dystonia. Overview of therapies that may alleviate symptoms. AB - Dystonia is an involuntary movement disorder characterized by twisting, turning, and posturing. This disorder may affect a single body part or may be more generalized, but the pathophysiology remains unclear. The treatment of choice for most of the focal dystonias is botulinum toxin injections, although oral medications occasionally may be beneficial. Surgical treatment of dystonia may be performed peripherally or centrally but is usually reserved for patients in whom other forms of therapy fail. PMID- 11043088 TI - Primary care guide to myoclonus and chorea. Characteristics, causes, and clinical options. AB - Myoclonus and chorea are hyperkinetic movement disorders that confer a jerky appearance. Myoclonus involves a quick and simple jerk, whereas the jerking in chorea combines with other, slower movements in a continuous, flowing fashion. Both disorders have many different causes, and diagnosis requires knowledge of common clinical characteristics and directed ancillary testing. Symptomatic treatment is available, but reversal of the underlying cause should be considered first if possible. The potential benefits of treatment must be weighed against the risk of drug side effects. PMID- 11043089 TI - Is it a tic or Tourette's? Clues for differentiating simple from more complex tic disorders. AB - Tics are characterized by sterotyped, purposeless, and irregularly repetitive movements and usually can be classified as chronic motor or vocal tic disorders, transient tic disorders, or Tourette's syndrome. The latter is a complex disorder associated with multiple tics and often accompanied by other conditions, such as ADHD and obsessive-compulsive disorder. Treatment can be difficult, and drug therapy should begin with agents least likely to cause problems for the patient. Education of the patient and family and support from the physician and other care providers are essential elements of effective management. PMID- 11043090 TI - Breast cancer diagnosis. Screening and evaluation. Institute for Clinical Systems Improvement. PMID- 11043091 TI - Can you name this congenital syndrome? Bardet-Biedl syndrome. PMID- 11043092 TI - Incidence, location, and diagnostic evaluation of metastatic bone disease. AB - Metastatic carcinoma is the most common malignancy of bone. The clinical presentation of patients with skeletal metastasis is variable. When asked to evaluate a patient with a pathologic lesion or unexplained bone pain, the orthopedic surgeon should follow a logical sequence of steps in evaluating the patient with suspected metastasis to optimize care and avoid complications. In the majority of cases, a systematic approach to the patient with skeletal metastasis leads to the correct diagnosis. PMID- 11043093 TI - The mechanism of metastasis. AB - The establishment of clinically detectable skeletal metastasis is a multifactorial process. This process can be divided into three general areas of understanding. The first is that of the intrinsic characteristics and properties of the tumor cells, which allow and facilitate their migration from the site of primary neoplasia to the distant host skeleton. Second, there are anatomic considerations of the human body, which influence the distribution of metastatic seeding. Third, there are the considerations of the host organism's biology, including the immune system, the circulatory system, and the affected host skeleton, which hinder and, at times, potentiate the ability of neoplastic cells to establish skeletal lesions. PMID- 11043094 TI - Pathology of skeletal metastases. AB - Metastatic disease involving the skeleton is an unfortunate and common occurrence in cancer patients. Choosing the best diagnostic approach requires knowledge of the patient's clinical history, the radiologic appearance of the lesion, the differential diagnosis, and the ability of the diagnostic modality to answer the questions that must be addressed. In difficult cases, interaction between the pathologist and clinician before biopsy may make the difference between a rapid procedure serving to definitively diagnose and effectively stage a patient and a costly procedure that provides little or no information. PMID- 11043095 TI - Medical management of metastatic skeletal disease. AB - The medical management of metastatic disease generally includes chemotherapy, hormonal therapy, and metabolic pharmacologic manipulations with medications, such as bisphosphonates as well as nonoperative physical measures, such as orthoses and ambulatory or mobility aids. This comprehensive complex care is best coordinated with the medical oncologist. If well planned and coordinated, such care can improve the life of the cancer patient greatly. PMID- 11043096 TI - Radiation therapy. AB - Radiation therapy is commonly used to alleviate the pain associated with bone metastases. This article reviews the components of the radiation oncology evaluation. The options for use of ionizing radiation including postoperative treatment, limited-volume external beam radiotherapy, wide-field radiotherapy, and radioisotope therapy are compared and contrasted. Side effects and toxicities of radiotherapy are discussed. PMID- 11043097 TI - Indications for operative treatment. AB - The importance of careful assessment of clinical, biomedical, and radiographic factors in the indications for operative treatment are stressed in this article. The indication for surgical resection of solitary lesions versus simple stabilization are compared and contrasted. The biologic behavior of various types of tumors is emphasized. PMID- 11043098 TI - Perioperative considerations in patients with metastatic bone disease. AB - Preoperative assessment of patients with metastatic bone disease includes a history and physical examination, laboratory evaluation, and standard radiographs. Perioperative diagnostics include technetium bone scan, CT scans, MR imaging, positron emission tomography, and biopsy. The role of preoperative tumor embolization and vena cava filter placement is discussed in this article. Guidelines for pain control are provided. Surgical planning and instrument considerations for long bone lesions, periarticular lesions, and pelvis and acetabular lesions are addressed. The importance of rehabilitation for patients with metastatic bone disease is emphasized. PMID- 11043099 TI - Management of metastatic lesions of the humerus. AB - The management of patients with metastatic disease in the humerus requires consideration of many factors. A careful evaluation of the patient's general condition, an understanding of the disease process, an appreciation of the degree of bone destruction, and a working knowledge of available treatment options are required. A multidisciplinary approach is essential to determine the course of treatment that best alleviates pain, preserves function, and optimizes the quality of life remaining in the patient with metastatic disease. PMID- 11043100 TI - Management of metastatic disease of the spine. AB - Accurate assessment of spinal cord stability, pain, and neurologic function is essential to any rational approach to metastatic disease of the spine. The Harrington classification is useful in terms of selecting treatment options and planning appropriate interventional and adjuvant therapy. For milder cases, with bone involvement in the absence of structural deformity, adjuvant therapy usually is appropriate. In cases of bone collapse, with or without neurologic compromise, surgical consideration should be given to patients with appropriate life expectancy. Surgical principles revolve around anterior decompression and reconstruction, for anatomic and biomechanical reasons. More widespread use of anterior instrumentation may obviate the need for subsequent posterior stabilization, but additional long-term study is required in this area. PMID- 11043101 TI - Operative management of metastases to the pelvis and acetabulum. AB - Bone metastases to the acetabulum and pelvis can be a devastating and debilitating problem. In certain patients, operative reconstruction of the involved hip can lead to maintenance of independence, pain control, and an increase in the overall quality of life for their remaining life span. These procedures are technically challenging and are associated with a higher complication rate than that for patients having surgery for nonneoplastic disease. They are probably best performed by surgeons with specific training and expertise in tumor surgery and acetabular reconstruction. With proper patient selection, appropriate component use, and competent surgical technique, good-to excellent results can be obtained. PMID- 11043102 TI - Metastatic disease of the femur. Surgical management. AB - Treatment of actual or impending pathologic fractures of the femur provides the senior author with some of the most rewarding surgical interventions of his practice. The patients' survival outlook is not changed, but their quality of life is enhanced significantly. Most health care providers usually provide the metastatic cancer patient only temporary symptomatic relief, at best, and often at the expense of continued pain, suffering, or sickness, such as is seen with chemotherapy-associated morbidity. Patients with metastatic bone disease are usually incredibly grateful for the restoration of function and diminution of their pain that results from the proper operation on metastatic bone disease. These patients typically are among the most appreciative patients and often express their gratitude when seen in follow-up in the clinic or office. Despite their metastatic disease state, their usual enthusiasm is uplifting to the surgeon and to the staff. To help a patient be pain-free and functional in the waning days of his or her life affords the patient, the physician, and the physician's staff with an emotionally rewarding experience and one that is well worth the time and effort required to care for these patients. By following the techniques outlined in this article, most patients with metastatic disease of the femur can be appropriately managed with excellent results. PMID- 11043103 TI - Management of metastatic disease of other bones. AB - Metastases to the scapula and distal sites on the upper and lower extremities are infrequent. Although these metastases tend to occur in patients with advanced disease, a distal metastasis is occasionally the sole metastatic location. Distal metastases do not pose an immediate threat to a patient's life; however, they may cause significant pain and disability. Appropriate management can considerably enhance function, quality of life, and, occasionally, survival. Seven cases of distal metastasis are presented in this article with discussion of operative and nonoperative approaches to management. PMID- 11043104 TI - Management of metastatic disease to soft tissue. AB - Metastases to soft tissue are rare clinical problems. Most metastases are caused by carcinomatous deposits in the skeletal muscle, with lung carcinoma being the most common primary cause. Pain is more commonly observed in association with metastatic soft tissue masses than for soft tissue sarcomas. Treatment should be individualized, but for most carcinomas, initial radiotherapy treatment is recommended. Prognosis varies with the underlying disease, but for the typical patient with a metastatic carcinoma, mean survival duration is approximately 6 months. PMID- 11043105 TI - Errors and pitfalls in the diagnosis and treatment of metastatic bone disease. AB - 1. The orthopedist must be sure of the diagnosis and not embark on treatment for the wrong diagnosis. 2. Solitary lesions in patients with a remote history of malignancy require complete investigation and biopsy. This includes blood work, bone scan, magnetic resonance imaging of the bone lesion, and CT scan of the chest and abdomen. 3. Pathologic fractures do not require immediate fixation. They require careful surgical planning and a team approach to the underlying malignancy. 4. Load-sparing devices should not be used. 5. Femoral neck fractures should be treated by endoprosthetic replacement, and consideration should be given to long-stemmed femoral components. 6. The orthopedist should assume that the fracture will never heal. 7. Immediate full and unrestricted weight bearing should be planned. 8. Future problems in the surgical site should be anticipated. Often a long-stem cemented femoral component is a better choice than a standard length. 9. The orthopedist must ensure that there are no other lesions that require stabilization in the bone being treated. 10. Methyl methacrylate can be used to augment fixation if needed. 11. If secure fixation cannot be achieved with the use of cement, the bone should be replaced with a tumor endoprosthesis. 12. The orthopedist should not hesitate to call in help. These can be difficult situations to manage and often require the assistance of a tumor surgeon and oncologic team. PMID- 11043106 TI - Weaving feminism and community psychology: an introduction to a special issue. PMID- 11043107 TI - A history of women and feminist perspectives in community psychology. AB - Using an historical framework, we document and assess efforts to include women, women's issues, and feminism in community psychology and in the Society for Community Research and Action (SCRA). Initiatives of the SCRA Task Force/Committee on Women are traced from its inception to present. We also chronicle the dilemmas and difficulties of moving toward a feminist community psychology. The history is divided into five phases. Each phase is described in terms of women's involvement in the field and efforts to integrate feminist content into research and practice of the field. Reflections on the qualities of contexts that have both supported and inhibited inclusion are identified. We look to this history to try to understand the observation that while women have been increasingly visible in leadership roles and women's professional development has been encouraged, less progress has been made toward building a feminist community psychology. PMID- 11043108 TI - The impact of welfare reform on men's violence against women. AB - Welfare reform is likely to have a profound effect on the lives of poor women who are being abused. This article proposes exchange theory and the feminist "backlash hypothesis" as frameworks with which to assess the impact of welfare reform on violence levels in abusive relationships. Exchange theory suggests that if a woman leaves welfare and obtains employment that increases her economic resources, violence against her will decrease. The backlash hypothesis makes a different prediction: Violence will increase as men attempt to compensate for women's enhanced status or independence. Both approaches are examined in light of current data. As demonstrated here, the incorporation into social policy analyses of feminist thinking about dominance and power will enrich our understanding of the impact of social policy changes on people. PMID- 11043109 TI - The importance of community in a feminist analysis of domestic violence among American Indians. AB - There are over 500 native communities in the United States alone. Although popular conceptions in the majority culture commonly refer to these as a single American Indian group, native communities are in fact extremely diverse and heterogeneous. Issues of gender, class, and power are discussed from a feminist perspective with an emphasis on the diversity among native communities. Available evidence, while sketchy, suggests that male authority, male restrictiveness, and socioeconomic stress are associated with violence, but that the levels of these factors vary widely across native groups. For example, some native tribes practice matrilineal descent while others are patrilineal. This diversity has far reaching implications for the community context in which domestic violence occurs. An approach that integrates both feminist and community approaches seems best suited to address the problem of domestic violence in native North America. PMID- 11043110 TI - Self-determination and empowerment: a feminist standpoint analysis of talk about disability. AB - In this paper we offer a feminist analysis of talk about self-determination and empowerment in the context of disability, focusing on the case of developmental disabilities. We find strains of the same patterns feminist epistemologists have argued shape the organization of formal knowledge from the standpoint of the privileged. At the extreme, people with developmental disabilities appear as objects without selves, outside of the context of interpersonal and social structural relationships that constrain who they can be by defining them as other, often in multiple and interacting ways. Empowerment, from the dominant standpoint, becomes an abstract attribute or condition; something a person has or does not have. Taking the standpoint of women and other marginalized people offers a view of self-determination as a person's development of his or her self. Empowerment becomes a potential characteristic of a social relationship, one that facilitates the development of someone's self. The most empowering relationships are mutual, recognizing and building on the diverse contributions and needs of participants in ways that seek to minimize inequalities over time. The reason some of us are self-determining is that we are in interpersonal and social structural relationships that empower us. To construct interpersonal and social structural relationships that empower people with developmental disabilities requires challenging the way dominant conceptualizations of independence and productivity also express the standpoint of the privileged. The standpoint of women allows all of us to talk more of how we connect with and facilitate one another's developing selves within communities. PMID- 11043111 TI - The Listening Partners program: an initiative toward feminist community psychology in action. AB - The Listening Partners intervention is described and analyzed as a synthesis of feminism and community psychology, within a developmental framework. Working from an empowerment perspective, this social action, peer group intervention supported a community of poor, rural, isolated, young, White mothers to gain a greater voice, claim the powers of their minds, and collaborate in developmental leadership--creating settings that promote their own development and that of their families, peers, and communities. High quality dialogue, individual and group narrative, and collaborative problem-solving were emphasized, in a feminist context affirming diversity, inclusiveness, strengths, social-contextual analyses, and social constructivist perspectives. The power of enacting a synergy of feminism and community psychology is highlighted. PMID- 11043112 TI - Where the girls (and women) are. AB - This paper takes up a theoretical and empirical investigation of how two community-based projects for young women both create safety from community and domestic violence but how, in the process, discourses of multicultural inclusion define one site, and racist discourses of exclusion float through the other site. By relying on two intensive qualitative case studies of community-based organizations for girls, one exclusively White and working class and the other expressly multicultural and antiracist, we try to identify those structures and practices that support feminist, but inadvertently racist, work and those structures and practices that enable, at once, feminist and antiracist consciousness and praxis. PMID- 11043113 TI - [Callus modulation--fiction or reality?]. PMID- 11043114 TI - [Mechanical modification of callus healing]. AB - Interfragmentary movement and size of the fracture gap influence fracture healing. Limited movements promote callus formation and may result in increased mechanical stability. Although larger movements still promote callus formation, the bony consolidation of the fracture is hampered. Fracture healing is also hampered if the size of the fracture gap is too large. A combination of large movement and large gap bears the risk of non-union. Therefore, having in mind a minimally invasive surgical approach, one should strive for good reduction of the fracture ends and flexible yet stable osteosynthesis. Dynamization of the fracture by enabling axial movement will close the fracture gap, stimulate tissue differentiation and possibly accelerate the healing process. External mechanical stimulation, however, has not been shown to effectively enhance the healing process under flexible fixation or in load-bearing patients. PMID- 11043115 TI - [Increasing expression of growth factors by adenovirus gene transfer. A new method of fracture treatment?]. AB - Gene therapy in orthopedic surgery is a new technic based on the idea of biological tissue healing. External gene segments are transferred to cells that overexpress growth factors locally to achieve this effect. The influence of growth factors on fracture healing is very well documented in the literature. Experimental data demonstrate that defect healing in bone can be accelerated by the application of different cytokines in vivo. Gene transfer is a promising, new technic of in situ tissue engineering that will enter clinics within the next decade. PMID- 11043116 TI - [Tissue engineering with mesenchymal stem cells for cartilage and bone regeneration]. AB - Tissue engineering offers the possibility to fabricate living substitutes for tissues and organs by combining histogenic cells and biocompatible carrier materials. Pluripotent mesenchymal stem cells are isolated and subcultured ex vivo and then their histogenic differentiation is induced by external factors. The fabrication of bone and cartilage constructs, their combinations and gene therapeutic approaches are demonstrated. Advantages and disadvantages of these methods are described by in vitro and in vitro testing. The proof of histotypical function after implantation in vivo is essential. The use of autologous cells and tissue engineering methods offers the possibility to overcome the disadvantages of classical tissue reconstruction--donor site morbidity of autologous grafts, immunogenicity of allogenic grafts and loosening of alloplastic implants. Furthermore, tissue engineering widens the spectrum of surgical indications in bone and cartilage reconstruction. PMID- 11043117 TI - [Recombinant growth hormone accelerates regenerate consolidation in distraction osteosynthesis]. AB - Distraction osteogenesis is a good model for evaluation of fracture healing with intramembranous bone formation. The purpose of the present study was to investigate whether homologous GH has a stimulating effect on bone healing in distraction osteogenesis. The left tibiae of 30 micropigs were osteomized and distracted with an external fixator 2 mm daily over a period of 10 days. Animals were killed after an additional healing time of 10 days. The treatment group received 100 micrograms r-pGH per kg bodyweight per day. A newly developed device allowed non-destructive torsional biomechanical evaluation of the regenerate strength as in vivo measurements. After killing, destructive torsional strength testing of the sites of distraction was performed. To determine the endocrine response to the administration of r-pGH, serum levels of IGF-I were determined. The non-destructive in vivo testing showed that torsional stiffness of the regenerate was significantly higher in the treatment group than in the control group. Final regenerate torsional failure load was 131% higher and ultimate torsional stiffness was 231% higher in the treatment group than in the control group. The mean serum level of IGF-I increased to 440% of the preoperative base level in the treatment group and remained unchanged in the control group. Our data indicate that systemic administration of recombinant homologous growth hormone significantly accelerates ossification of bone regenerate in distraction osteogenesis. PMID- 11043118 TI - [Local liberation of IGF-I and TGF-beta 1 from a biodegradable poly(D,L-lactide) coating of implants accelerates fracture healing]. AB - In vitro and in vivo studies have demonstrated an osteoinductive effect of growth factors like IGF-I and TGF-beta 1. However, for therapeutic use in fracture treatment, the local application of these bioactive molecules is still an unsolved problem. The controlled release of growth factors from a biodegradable coating of osteosynthetic implants could stimulate fracture healing locally. Coated implants could stabilise the fracture and work as a local drug delivery system. Previous studies demonstrated a high mechanical stability of a thin 10 microns poly(D,L-lactide) (PDLLA) coating on metallic implants that withstands even an intramedullary insertion process. After an initial peak, 80% of incorporated growth factors IGF-I and TGF-beta 1 were continuously released within 42 days. The effect of locally applied IGF-I and TGF-beta 1 from a biodegradable PDLLA coating of intramedullary implants on fracture healing were investigated in a rat model. A fracture of the right tibia of 5-month-old female Sprague-Dawley rats was stabilised with coated versus uncoated titanium K-wires. X-ray examinations and blood analysis were performed, body weight and body temperature monitored throughout the experimental period. After 42 days both tibiae were dissected for mechanical torsional testing and histomorphometric analyses. The results demonstrate a nearly completely consolidated fracture in the X-ray examinations, a significant higher maximum load and torsional stiffness in the biomechanical tests and a progressed remodeling in the histological and histomorphometric analyses after 42 days in the group treated with growth factors compared to the controls. Interestingly, the PDLLA coating itself had a positive effect on fracture healing even without incorporated growth factors. No systemic change of serum parameters including IGF-I and IGF binding proteins and no differences in body weight and body temperature were seen in any group. These findings suggest that the local application of growth factors from a biodegradable poly(D,L-lactide) coating of osteosynthetic implants accelerates fracture healing significantly without systemic side effects. PMID- 11043119 TI - [Surgery of the cruciate ligament--an ever current topic]. PMID- 11043120 TI - [New methods of cruciate ligament surgery]. AB - Although long-term results in cruciate ligament surgery are continuously improving, we are still confronted with a certain rate of surgical failures and an increasing number of revision procedures. Despite a tremendous effort towards experimental and clinical research in the field of anterior and posterior cruciate ligament surgery over the past decades, there is still a strong need to further improve results. Therefore, the goal of the present paper is to introduce and discuss new developments in the rapidly changing field of cruciate ligament surgery, in order to give the clinician a base for discussion to include these developments in their daily clinical work. The present paper focuses on current developments and future perspectives such as biodegradable implants, new aspects in graft selection, double-bundle posterior cruciate ligament reconstruction, possibilities of biological intervention with growth factors, and the new technology of robotics and navigation. PMID- 11043121 TI - [Current aspects of anchoring hamstring tendon transplants in cruciate ligament surgery]. AB - The use of hamstring tendon grafts in cruciate ligament surgery has recently raised strong interest. Hamstring tendons are superior to the mid third patellar tendon graft by virtue of lower harvest site morbidity combined with high tensile strength. Osseous graft incorporation relies on a proper tendon-to-bone healing, which relies on specific biomechanical and biological boundary conditions. Several different fixation devices have recently been introduced, with special emphasis on high initial fixation strength and moving the level of fixation closer to the joint line, the so-called aperture fixation. The goal of the present review is to focus on the advantages and disadvantages of different fixation principles for hamstring tendon grafts in order to give a comprehensive insight into current developments, such as interference fit fixation, cross-pin fixation, and the concept of hybrid fixation. PMID- 11043122 TI - [Differential transplant selection in cruciate ligament surgery]. AB - Over the past century numerous graft materials have been used for the reconstruction of the cruciate ligament of the knee. Among the autologous tissues that are currently recommended as graft materials, the central bone patellar tendon bone graft, a quadrupled hamstring graft and the central quadriceps tendon graft have the greatest clinical significance. With some limitations, allograft materials can also be used. Each of the three mentioned grafts has specific features regarding morphological and structural properties, graft fixation and graft incorporation. Clinical studies have failed to identify any of the three grafts as superior to the others. When choosing the graft for surgery the different anatomy and function of the anterior and posterior cruciate ligaments have to be considered. For the treatment of multiple ligament injuries and for revision cases, thorough preoperative planning is necessary and modified graft selection may be required. PMID- 11043123 TI - [Strategies for interventional revisions in failed anterior cruciate ligament reconstruction]. AB - Anterior cruciate revision reconstruction is gaining more and more importance. Postoperative infection, a painful knee, limited range of motion and instability may make a second operation necessary. Results after revision ACL reconstruction are worse than results after primary ACL reconstruction. Analysis of the causes of failure and a therapeutic concept that is tailored to the individual case are preconditions for a successful reintervention. Revision ACL reconstruction has to be performed by an experienced knee surgeon who masters all the necessary techniques, from arthroscopy to arthrotomy and should be carefully planned. PMID- 11043124 TI - [Diagnosis and therapy of fresh and chronic posterior cruciate ligament lesions]. AB - Injuries to the posterior cruciate ligament are among the major injuries to the knee joint. Today we are often confronted with an inappropriate and delayed diagnosis of this injury and its concomitant lesions. Additionally, the outcome of operative treatment has not yet reached an acceptable rate of satisfactory results. Therefore, the goal of this current concept review is to give a comprehensive insight into anatomy and biomechanics, injury mechanisms and pathobiomechanics, based on our experiences and data from the international literature. A further goal is to clarify diagnostic problems with respect to clinical examination and imaging techniques. We also present a differential concept for the perioperative and conservative management of posterior cruciate ligament deficient knees in order to also adequately address concomitant injuries such as posterolateral rotatory instability and combined anterior cruciate ligament injuries with the aim of further improving results. PMID- 11043125 TI - [Conservative or surgical therapy of acromioclavicular joint injury--what is reliable? A systematic analysis of the literature using "evidence-based medicine" criteria]. AB - There is controversy about the therapy for third-degree acromioclavicular dislocation according to Tossy and Rockwood's classification. Both operative and non-operative treatment is reported to have satisfactory results in the literature. The purpose of this study was to analyze the literature in a systematic manner based on the criteria of evidence-based medicine. It was our hypothesis that there is no scientific evidence for the superiority of one treatment over the other. A total of 370 papers were retrieved and classified into three groups: (1) randomized controlled trials; (2) comparative retrospective studies; and (3) retrospective studies. In three studies that were graded with high evidence, the major outcome for both operative and non-operative treatment was similar. The advantages of non-operative treatment include a shorter period of rehabilitation and a significantly lower complication rate while the advantages of operative treatment include a low rate of persisting subluxation of the AC joint. Similar results were found for retrospective comparative and long-term studies. For retrospective studies without controls, both operative and conservative therapy are described with good and excellent results, ranging between 80 and 97%. In conclusion, there is good evidence on the therapy of third-degree acromioclaviculary dislocation studies. The functional result according to the literature is similar, and complications associated with therapy occur more often with operative treatment. Conservative treatment appears to be the method of choice for third-degree acromioclavicular dislocations unless the patient's preference is operative therapy. PMID- 11043126 TI - [Osteochondral transplantation versus autogenous chondrocyte transplantation. A prospective comparative clinical study]. AB - INTRODUCTION: The treatment of full-thickness cartilage defects still represents a problem that has not yet been solved satisfactorily. Current methods used to cover defects in the knee joint are osteochondral cylinder transplantation (OCT) and autologous chondrocyte transplantation (ACT). METHODS: With a prospective clinical investigation, at the time being with 2-year results, we have examined ACT in comparison to OCT in 20 patients with regard to clinical and histomorphological (histology, immunohistochemistry, RES) outcome. RESULTS: We found equally good results with both methods in Lysholm, Meyers and Tegner Activity Scores. Histomorphologic evaluation of biopsies obtained by arthroscopy after ACT showed a defect filling in all cases, mainly with fibrous cartilage, while localized areas of hyalinelike regenerative cartilage were documented near the base. We did not see any histomorphologically visible change in the transplants after OCT. CONCLUSION: At the time we prefer OCT instead of ACT given the correct indication. PMID- 11043127 TI - [Diaphyseal femur pseudarthroses--only a technical problem?]. AB - Between 1981 and 1994 at the Bergmannsheil Ruhr University Hospital in Bochum, Germany, we treated 145 patients with femoral diaphyseal nonunions following initial operative treatment. Of these patients, 138 received this initial operative treatment at an outside institution. The primary reconstructions for the fractures utilized plates in 112 cases, reamed nails in 24 cases and external fixators in 9 cases. The average age of the patients was 35 years and the mean time from the initial operative treatment was 2 years. Twenty-seven patients (19%) presented with a hypertrophic nonunion and 118 (81%) with an atrophic nonunion. There was a significant correlation between primary "classic" plating and development of an atrophic nonunion (chi 2-test: P < 0.01). We observed 34 wound infections (23%) with no significant correlation to the type of primary osteosynthesis. We determined that 73 of the pseudarthroses were due to improper osteosynthesis techniques. Of these cases, 41% involved the use of plates, 83% involved the use of reamed nails, and 78% involved the use of external fixators. Fracture location near the diaphyseal-metaphyseal junctions was common in this problematic group. Ninety-two percent of all plates led to atrophic nonunions. There were 21 open fractures and of these 90% (n = 19) developed an atrophic pseudarthrosis and 29% (n = 6) developed a wound infection. Fifty-seven (39%) of all patients had additional injuries, but we found that did not increase the risk of disturbed bone healing. Our revision operations focused on the elimination of wound infections, refreshment of bone healing, and improvement in fragment stability. Only 28% of all "classic" plates and 11% of all external fixators were changed to an intramedullary implant at the time of the first revision surgery. Hypertrophic nonunions required a mean of 1.3 revision operations to achieve bone healing whereas a mean of 2 revision operations were necessary to fuse atrophic bone ends (P < 0.05). In cases of diaphyseal pseudarthrosis healing time was not affected by the type of osteosynthesis used for primary reconstructions. Since lack of fracture healing can often already be observed directly from postoperative X-rays, we recommend that revision procedures be performed early. The prolonged length of time to care for femoral nonunions underlines the importance of appropriate primary fracture treatment. That takes into consideration both the biomechanical and the biological aspects of bone healing. PMID- 11043128 TI - [The long gamma nail--indications, technique and results]. AB - The introduction of the Gamma nail (GN) as an intramedullar implant for pertrochanteric femoral fractures that allowed full weight bearing decreased the death rate from 17% (methods without full weight bearing) to 6%. The long Gamma nail (LGN) is a logical supplement of the standard version, designed to treat unstable per-, subtrochanteric and segmental fractures. This study evaluated 44 consecutive operations. Seventy percent of the patients had to be classified ASA III and IV, due to their high morbidity. The median age was 73.5 years. Multiple injuries occurred in 30.2%. All fractures were considered unstable. Surgery was usually performed within 24 h. The median duration of the surgical treatment was 120 min. In five cases technical problems were observed. Radiological controls showed a good positioning of the head screw. Early complications consisted of four local wound infections, three of them deep infections with a osteomyelitis. Deep venous thrombosis was observed in four cases, two of which included a pulmonary embolism (conservative treatment). The 30-day death toll was 2.3% (one patient). The median survival time (using Kaplan-Meier) in the study was 46 months, compared to 80 months in a matched population. This difference has to be linked to high premorbidity. The median duration of admission was 15 days. Mobilisation with full weight bearing was theoretically possible in all cases, but additional injuries or preoperatively impaired walking ability prevented full mobilisation in 15 cases. Functional assessment uncovered a decrease in Merle d,Aubigne score of 26.7% due to an impaired walking ability. Seventy-three percent of the patients regained their preoperative social status. In conclusion the long Gamma nail is a universal, less invasive implant with high early weight bearing. It thus allows early remobilization and reduces lethality in the treatment of complex, unstable coxal fractures. PMID- 11043129 TI - [Chronic venous insufficiency after open tibial fracture. An underestimated problem]. AB - There is a wide range of alternatives for primary bone reconstruction in the treatment of open lower leg fractures with soft tissue damage of the type Gustillo II and III. The primary objective should always be the protection of soft tissue damage whether one uses the fixateur externe, or an unreamed nail or primary bone shortening with secondary callus distraction. In recent years, this approach has produced better results and a reduction in the rate of major amputations. The overall effects of the initial treatment can only be analysed after a number of years. Research results indicate long-term soft tissue complications of the lower leg, varying from harmless swelling to venous ulcer. We conducted a clinical investigation which compared 80 patients, who were treated between 1985 and 1994 using the venous-occlusion plethysmography, to 50 healthy individuals. Clinically significant damage of the deep venous system was found in over 50% of cases. There was a direct correlation between the number of years since the initial treatment and the degree of damage found. Based on these findings, we recommend that the initial treatment of this condition and the preventative treatment of the secondary trauma diseases should follow regulated surgical guidelines and be recognised for insurance purposes. PMID- 11043130 TI - [Pathological fracture in primary malignant bone tumors]. AB - Retrospective analysis of 30 patients with pathological fracture out of 336 patients with primary malignant bone tumors should demonstrate the influence a pathologic fracture and the form of surgical therapy have on the survival rate. In 25 out of 30 patients a fracture led to diagnosis of the disease. Pathological fractures occurred cumulatively by malignant fibrous histiocytoma of the bone and in tumor stages IIb and III. Surgery was performed on 26 out of 30 patients (12 ablative therapies, 14 reconstructive therapies). The mortality risk for patients with pathological fractures was more than double the risk for patients without pathological fractures (P = 0.0062). When performed correctly, reconstructive therapy does not influence the survival rate. PMID- 11043131 TI - [Does liberation of interleukin-12 correlate with the clinical course of polytraumatized patients?]. AB - INTRODUCTION: Interleukin-12-p70 (IL-12-p70) is a potent immunoregulatory cytokine composed of a heavy chain (p40) and a light chain (p35). Contradicting results have been reported with regard to leukocyte release and systemic concentration of IL-12 after polytrauma. METHODS: We daily analyzed systemic concentrations of IL-12 in polytrauma patients (n = 37, mean ISS 33.9) in comparison to healthy donor values during intensive care course by ELISA. Patients were divided according to their mean IL-12 levels into those with elevated IL-12 (group 1, n = 7), those with decreased IL-12 (group 2, n = 4) and those with IL-12 in the normal range (group 3, n = 26). RESULTS: Patients in group 1 revealed elevated levels of IL-12 up to p70 > 1000 pg/ml and p40 > 2500 pg/ml. The common clinical feature of group 1 was a thorax trauma in combination with pneumonia (85% survivors). Patients with single thorax trauma or pneumonia without thorax trauma (group 3) showed normal IL-12 values. Patients with decreased IL-12 levels revealed also a thorax trauma and pneumonia but all patients succumbed. The groups significantly differ in their stay in the intensive care unit, in TISS, in MODS score and in respiratory ratio, but not in ISS, mean CRP values and leukocyte counts. Correlation analysis revealed no significant relation between systemically altered IL-12 values and clinical parameters, with the exception of a negative correlation of p70 and ISS (r = 0.785) or MODS score (r = -0.314) in group 1. CONCLUSIONS: After major injuries there is no overall suppression of IL-12 formation. Patients with normal or elevated IL-12 levels belong mainly to the survivors, whereas patients with decreased IL-12 levels are at high risk of succumbing to multi-organ failure. PMID- 11043132 TI - [Determinants of the global quality of life after polytrauma]. AB - This study investigated multiple trauma patients, who were injured between 1991 and 1995 and treated in our department. The aim of this study was to identify the determinants of quality of life after multiple trauma. From a total of 186 patients 173 (93%) were examined. The patients were asked to rate their quality of life according to the Nottingham Health Profile (NHP) and to a visual analogue scale (VAS). The VAS and the NHP isolated the age of the patients, the duration of artificial respiration, and the duration of rehabilitation as the predictors for a reduced overall quality of life. These results show that quality of life after multiple trauma not only depends on the severity of injury but also on demographic and psychosocial factors. PMID- 11043133 TI - [Penetrating injuries of the neck, injury pattern and diagnostic algorithm]. AB - Penetrating neck injuries are potentially life-threatening injuries. Management is controversial despite decades of discussion in the literature. An algorithm for diagnosis and therapy is needed because of the potential risk of mortality. In the Department of Trauma Surgery of Vienna Medical School, mandatory surgical exploration after noninvasive investigations is practiced. Thirty-one patients with this kind of injury were treated between August 1992 and September 1999. Injuries were caused in seven cases by gunshots, in eight cases by broken glass, in one case by an iron rod, in one case by a spear, in one case by a branch and in 13 cases the penetrating injuries were stab wounds caused by knives. Complications like pseudoaneurysms and fistulas were not seen. No patient came to death. PMID- 11043134 TI - [Tumor surgery of the upper cervical spine]. AB - Our own series of tumors of the upper cervical spine was analyzed retrospectively. The standard treatment strategies were reevaluated. A total of nine patients (mean age 61 years, metastasis 4, plasmocytoma 3, chordoma 1, histiocytosis 1) were treated between 1/92 and 2/99. A total of 12 operations were carried out. One-step procedures (6): Three extraoral, one transoral, one dorsal and in one case combined dorsal and extraoral tumor removal were performed. Three dorsal occipitocervical or atlantoaxial stabilizations, one ventral plating and two combined ventral plating plus dorsal three-point fixations, and four vertebral body replacements were carried out. Two-step procedures (3): three extraoral tumor removals with ventral plating plus dorsal three-point fixation, in combination with vertebral body replacement in two cases. The neurological status and the quality of life (Karnofsky performance status, pain levels) were analyzed preoperatively and at the follow-up outpatient examinations (mean follow-up: 18 months). Flexion-extension radiographs were performed at the follow-up. There was no operative mortality. The transient morbidity was 11%. The operative intervention improved the quality of life in all patients. Three patients died within 27 months of operation. Tumor resection at the upper cervical spine using individually modified surgical strategies over an approach corresponding to the tumor location, stabilization and vertebral body replacement increases significantly the time of survival and quality of life with an acceptable surgical risk for complications. PMID- 11043135 TI - [Therapy and prognosis of enchondroma of the hand]. AB - Enchondromas are the most common bone tumors of the hand. In a retrospective study, medical records and radiographs of 112 patients were reviewed. These patients were operated on between January 1973 and June 1997. After extirpation of the tumor, the defect was preferably treated with bone grafting in 102 patients. A malignant transformation (chondrosarcoma) was diagnosed in 2 patients. Follow-up examination of 92 patients with a mean follow-up of 1.6 years (range: 7 months to 14 years) yielded excellent or good results in 76 patients (82.6%), according to the evaluation scheme of Wilhelm and Feldmeier. Four recurrences were probably related to an incomplete resection of the tumor. The authors conclude that enchondromas of the hand should be treated surgically to prove the diagnosis and to prevent a pathologic fracture. PMID- 11043136 TI - [Plastic surgery coverage of osteocutaneous defects of the sternum area with the vertical and transversal rectus abdominis muscle (VRAM/TRAM) flap]. AB - Longitudinal osteocutaneous defects of the sternal region including the caudal third were reconstructed in 15 patients during a 3-year period by using the "vertical (VRAM)- and transverse rectus abdominis muscle" (TRAM) flap. The majority of the defects resulted from chronic osteomyelitis after previous cardiothoracic surgery or were due to former therapy of breast cancer. Three VRAM/TRAM flaps were primarily transferred as free flaps with microvascular anastomosis in the axilla region. Nine out of 12 pedicled VRAM or TRAM flaps required an additional microvascular anastomosis because of imminent venous or arterial insufficiency ("supercharging"). Therefore, operative technique and operating time of the pedicled and free flap for reconstruction of longitudinal sternal defects are comparable. Adequate reconstruction and rehabilitation was achieved in 11 cases. In 2 patients revision and partial secondary defect coverage was required. Two male patients died postoperatively due to their preexisting condition. PMID- 11043137 TI - [Covering soft tissue defects and unstable scars over the Achilles tendon by free microsurgical flap-plasty]. AB - INTRODUCTION: Coverage of the exposed Achilles tendon requires thin, supple tissue to provide adequate range of motion and a satisfying aesthetic result for the distal lower extremity. Various local flaps and free flaps have been described for reconstruction of small and large defects. Small defects can be closed with local tissue, whereas free flap coverage may be necessary for coverage of large defects. METHODS: From July 1993 to September 1998 14 patients between the age of 15 and 74 years (mean 47 years; 3 female, 11 male) underwent free flap coverage for the exposed Achilles tendon due to primary trauma, chronic wounds or tumors. The mean duration of follow-up was 33.3 months. The defect size ranged from 8 x 8 to 25 x 28 cm. RESULTS: Six parascapular flaps (three with a vascularized scapular fascial extension), four radial forearm flaps and four latissimus dorsi flaps (one combined with free serratus fascia) were used for soft tissue coverage over the Achilles tendon. Thirteen flaps survived. In one case a parascapular flap had to be removed due to venous thrombosis and a free latissimus dorsi flap was used as secondary salvage procedure. The donor site morbidity was acceptable for most patients after flap harvesting in the subscapular region and also satisfactory in the forearm region. Average active range of motion in the upper ankle joint was 15-0-40 degrees for extension/flexion. All patients were satisfied with the functional and aesthetic result. CONCLUSION: Soft tissue coverage over the exposed Achilles tendon requires an optimal solution for each patient to achieve an aesthetically pleasing result and acceptable function. Microvascular free flaps can be used to reconstruct medium and large defects and to provide gliding tissue for the Achilles tendon. The complication rate of microvascular flaps is comparable with that of local flaps. PMID- 11043138 TI - [A new technique of subdermal flap dissection for reconstruction of the ala of the nose]. AB - There are different techniques for reconstruction of the ala of the nose. Nasolabial flaps are commonly used. With these techniques nasolabial and perialar skin is subdermally dissected and turned over into the defect to create both the inner and outer lining of the ala. If there is also loss of the perialar and nasolabial skin, such techniques become impossible. With a modification it is still possible to reconstruct both the ala of the nose and the perialar/nasolabial skin in one operation. A V/Y sliding flap from the cheek with a side extension from the lower nasolabial fold used as a turn-over flap is dissected. With this technique it is also possible to reconstruct isolated loss of the ala of the nose. PMID- 11043139 TI - [Trans-scaphoid perilunar dislocation of the wrist (de Quervain) as a rare complication of electric injury]. AB - We describe the case of a 67-year-old man with an electricity inflicted injury in the left hand and a transscaphoidal perilunear dislocation of the opposite right hand. On admission, the injury was missed on the standard ap-radiogram of the right hand, so the operative reconstruction was delayed. Later on, the lunate showed aseptic necrosis and wrist collapse as consequence. PMID- 11043140 TI - [Comment on H. J. Oestern: Distal radius fractures. 1: Basic principles and conservative therapy]. PMID- 11043141 TI - [Value of flexible endoscopy in surgery. I]. PMID- 11043142 TI - [Ambulatory surgery in the hospital (section 115b SGB V)]. PMID- 11043143 TI - [Effects of the "law for controlling corruption" on the relations been physicians and the health care industry]. PMID- 11043144 TI - [BDC (Professional Society of German Surgeons) online. New internet presence of the professional society]. PMID- 11043145 TI - [The Delegation of the Current Professional Group of Young European Physicians (PWG) in the Committee for Surgery of the European Union of Specialists (UEMS)]. PMID- 11043146 TI - [Virus carrier in surgery]. PMID- 11043147 TI - Adverse cardiovascular drug interactions. PMID- 11043148 TI - Survey of national guidelines for the treatment of phenylketonuria. AB - Phenylketonuria treatment policies vary not only between different countries worldwide, but also within one country. Recommendations and guidelines for phenylketonuria should deal with the following subjects: 1. What is the target age to start dietary phenylalanine restriction under newborn-screening conditions? 2. At which plasma phenylalanine concentration should phenylalanine restriction be initiated? 3. Which are the recommended plasma phenylalanine concentrations at different ages? 4. What is the recommended frequency of monitoring phenylalanine in plasma? Statements from the following countries are presented: Czech Republic, Denmark, France, Germany, Great Britain, Hungary, Ireland, Poland, Slovakia and the United States. CONCLUSION: Due to the lack of internationally accepted guidelines, management of phenylketonuria still varies between different countries. Our efforts should focus on the formulation of internationally acceptable and accepted recommendations for the treatment of patients with phenylketonuria at different ages. PMID- 11043149 TI - Development of intelligence in early treated phenylketonuria. AB - Designs, analyses and results of longitudinal studies of intelligence of patients treated early for phenylketonuria (PKU) are reviewed. All studies converge on the conclusion that after the age of 10 years, IQ development is stable for different degrees of dietary relaxation. On average, for each 300 mumol/l increase in blood phenylalanine (Phe) levels pre-school, IQ decreases by about half a standard deviation. Children with Phe levels below 400 mumol/l in early and middle childhood had the best outcomes which were near normal. PKU seems to suppress the global level of IQ without impairment of domain-specific competencies. For historical reasons there is no research on IQ development of early treated patients in middle or late adulthood, and it remains unclear whether older age groups might carry new risks. CONCLUSION: It is argued that control group designs, meta-analysis, and interdisciplinary studies combining psychology, neurology and neuropathology could increase the understanding of phenylketonuria as well as the scientific basis of its treatment. PMID- 11043150 TI - Comments on cognition and intelligence in phenylketonuria. PMID- 11043151 TI - Neuropsychological approaches to treatment policy issues in phenylketonuria. AB - Neuropsychological research conducted so far on treatment factors in phenylketonuria suggests that dietary cessation at age 6 is too early. However, continuation of diet until age 10 appears to provide protection against subsequent hyperphenylalaninaemia in the domains of perception, memory and motor skill if concentrations remain at least below 1200 mumol/l thereafter. Levels in the range 360 mumol/l to 600 mumol/l appear to constitute a hazard for executive skill during the pre-school period but in the primary school years the risk diminishes. Levels above 900 mumol/l in early adolescence and adulthood may affect executive abilities adversely and the question remains whether such effects are reversible. CONCLUSION: Though scant, neuropsychological evidence does not seriously contradict current British and German recommendations for dietary control in phenylketonuria. PMID- 11043152 TI - Behaviour in early treated phenylketonuria: a systematic review. AB - A search was made of the literature on behaviour in early treated phenylketonuria (PKU). Some 34 publications were selected for systematic review in order to answer the following questions: (1) does behaviour in early treated subjects with PKU differ from that in the general population?, (2) if so, do the differences show any consistent pattern?, matter in real life?, relate to intellectual status, phenylalanine (Phe) control or social factors? and (3) can behavioural problems be precipitated or reversed by changes in Phe control?. Eleven reports based on four studies of > 20 subjects provided the most useful data. There was good evidence of differences between subjects with PKU and their unaffected peers in behaviour and perception of self. The pattern of results was consistent across nations and age groups (higher scores on emotional/neurotic items and lower scores on anti-social/aggressive items) and suggested that subjects with PKU are more prone to depression, anxiety, phobic tendencies, isolation from their peers and a less "masculine" self image. Features of attention deficit hyperactivity disorder and associations between behaviour problems and both lower IQ and a worse quality of Phe control were well documented in several studies. CONCLUSION: The literature did not allow for definite conclusions on the cause of the behavioural problems but, taken together, the findings suggest that both the stressful dietary intervention and a degree of neurobiological impairment are likely to play a part. PMID- 11043153 TI - Patterns of academic achievement among patients treated early with phenylketonuria. AB - This paper provides a review of recent studies investigating the academic achievement of patients treated for phenylketonuria. These studies consistently indicate that these patients demonstrate mild academic difficulties, primarily with mathematics but with reading and spelling skills as well. In addition to the review of previous studies, academic achievement results are presented for a sample of 32 school-age, early-treated patients with phenylketonuria. CONCLUSION: Consistent with previous studies, results from the current study suggest that patients in this sample demonstrate mild difficulties with basic spelling and basic mathematical calculation skills. PMID- 11043154 TI - The neuropathology of phenylketonuria: human and animal studies. AB - Pathologic changes in the brain of untreated phenylketonuria (PKU) patients occur in structures that develop post-natally, i.e. in myelination of subcortical white matter and spinal cord and in the growth of axons, dendrites and synapses in cerebral cortex. In addition, a small minority of brains show evidence of progressive white matter degeneration (leucodystrophy). The pathologic changes are thought to be due to toxic effects of phenylalanine and/or its metabolites. It is assumed that they can be prevented by dietary therapy during infancy and childhood, but direct confirmation by neuropathologic studies is lacking. The recently discovered genetic mouse mutant Pah(enu2) provides an excellent animal model in which effects of PKU on brain development, including dendritic and synaptic development in cerebral cortex, can be assessed. In human PKU, there needs to be neuropathologic study of the brains from PKU patients, particularly adults, with a history of dietary therapy. Special attention needs to be paid to the study of white matter in such cases, in view of recent reports of white matter lesions on MRI despite dietary treatment. CONCLUSION: Careful correlation is needed between neuropathology, magnetic resonance imaging white matter changes, dietary history and clinical findings. Finally, neuropathologic investigation is needed to determine whether progressive degeneration of the white matter (leucodystrophy) poses a risk to adults in whom dietary therapy has been discontinued. PMID- 11043155 TI - Comments on the neuropathology of phenylketonuria. PMID- 11043156 TI - The neurochemistry of phenylketonuria. AB - The mechanisms by which deficiency of hepatic phenylalanine hydroxylase causes central nervous system disease are reviewed. The neurological disease appears to be secondary to increased concentrations of phenylalanine and a decrease in the concentrations of other large neutral amino acids, especially methionine and tyrosine, within the central nervous system. This causes a deficiency of the neurotransmitter dopamine, reduced protein synthesis and demyelination. Similar mechanisms appear to be operating when blood phenylalanine concentrations are in the range expected for early continuously treated phenylketonuria. CONCLUSION: The severe brain disease found in phenylketonuria is caused by a raised blood phenylalanine content which increases the brain free phenylalanine and decreases the concentration of other large neutral amino acids. Brain protein synthesis is decreased, myelin turnover is increased and there are abnormalities in amine neurotransmitter systems. PMID- 11043157 TI - Adult care in phenylketonuria and hyperphenylalaninaemia: the relevance of neurological abnormalities. AB - Neurological abnormalities in phenylketonuria were described before dietary treatment became possible. These included tremor, clumsiness, epilepsy, spastic paraparesis and occasionally extrapyramidal features. Neurological deterioration after childhood was recognised. Patients with neurological deterioration described recently have been late diagnosed or intellectually impaired or both. No early diagnosed patient who was well treated and of good intellectual outcome has yet shown neurological deterioration after stopping diet but it may happen. CONCLUSION: The fascinating links between pathology, magnetic resonance imaging appearances, magnetic resonance spectroscopy results and clinical features are not yet clearly understood. Patients must understand the possible risks of stopping diet and make their choice. All patients need help, support and follow up regardless of the choices they make over continuing diet. PMID- 11043158 TI - In vivo proton magnetic resonance spectroscopy in phenylketonuria. AB - In vivo nuclear magnetic resonance spectroscopy permits the non-invasive examination of metabolic characteristics of the human brain in a clinical environment. Methods to detect elevated phenylalanine (Phe) in patients with phenylketonuria (PKU) using difference spectroscopy and to estimate absolute brain Phe concentrations, [Phe]brain, have been developed. In patients with classical PKU, [Phe]brain typically varied between 0.14 and 0.78 mmol/l depending upon actual blood Phe concentrations, [Phe]blood, between 0.47 and 2.30 mmol/l. Dynamic investigations can be used to extract information about Phe transport at the human blood-brain barrier, which may be described by a symmetric Michaelis Menten model. Carrier saturation and competitive inhibition of the influx of other large neutral amino acids can be expected at blood levels usually found in PKU patients. In single cases of untreated, normal intelligent patients, abnormally low [Phe]brain < or = 0.15 mmol/l were observed despite high stationary Phe levels ([Phe]blood = 1.15 +/- 0.10 mmol/l). CONCLUSION: Significant variations in phenylalanine transport parameters in untreated, normal intelligent patients indicated that blood-brain barrier transport or intracerebral phenylalanine consumption are causative factors for the individual vulnerability to phenylketonuria. PMID- 11043159 TI - Nutrition, physical growth, and bone density in treated phenylketonuria. AB - Dietary treatment of phenylketonuria is well established to be safe and to prevent developmental and mental impairment in patients with low or absent phenylalanine hydroxylase activity. The use of semi-synthetic diets necessitates careful and longitudinal control not only of physical and intellectual development, which are both near normal in well treated patients, but also of potential diet inherent insufficiencies of essential nutrients. Concern has been raised by some reports on growth retardation in young patients on strict diets and on decreased bone density in older phenylketonuric children. The clinical significance of these findings is not known. CONCLUSION: Changes have been found, although inconsistently, in connection with selenium, zinc, iron, retinol and polyunsaturated fatty acid status in dietetically treated patients with phenylketonuria. Both the mechanism and significance of these changes is doubtful at present. PMID- 11043160 TI - Diet and compliance in phenylketonuria. AB - In phenylketonuria, compliance and diet is a difficult issue; it is hard to quantify and is under-researched. It is influenced by many factors. Failure to consume prescribed quantity of protein substitute has been commonly reported and is probably affected by their acceptability, format and timing of administration. There are few reports documenting actual phenylalanine intake and blood phenylalanine control, but it is possible that the more rigorous systems for allocation of phenylalanine are associated with worse compliance. The dry, hard and insipid nature of many low protein products may lead to their under usage and consequent boredom and hunger. The diet becomes increasingly harder to maintain as children grow older and seek fewer constraints in their meals. CONCLUSION: In phenylketonuria, encouraging adherence to diet requires continual education, reinforcement and support from the family and professionals within the support team. PMID- 11043161 TI - Final intelligence in late treated patients with phenylketonuria. AB - Despite neonatal screening programmes, there is still a number of patients with phenylketonuria who are not diagnosed and start treatment late. The question in this study was to evaluate which factors will contribute, other than the quality and duration of dietary treatment, to final outcome in late treated patients with phenylketonuria. We retrospectively analysed the data of 40 patients with phenylketonuria, of whom 2 patients at 35 and 24 years of age had a normal IQ despite never being treated. In 38 patients starting dietary treatment between 0.7 and 7 years of age, mean IQ/DQ at diagnosis was 52.7 (SD = 16) (mean age 2.5 years), final IQ (mean age 33.5 years) was 79.0 (SD = 16), the difference was highly significant (P < 0.0001). Important factors for the final intelligence in adult late treated patients with phenylketonuria were onset (r = -0.46, P < 0.009) and DQ/IQ (r = 0.51, P < 0.002) when dietary treatment was started. Thus, in late treated patients with phenylketonuria, in addition to the quality and duration of treatment, the outcome is mainly influenced by the age of starting treatment and also by the intellectual status of the patient. In one of the two patients with normal intelligence, nuclear magnetic resonance spectroscopy showed that brain phenylalanine was undetectable even though blood phenylalanine was 30 mg/dl. A second metabolic disorder may protect these patients from severe brain damage. CONCLUSION: These data indicate that brain damage in untreated or late treated patients with phenylketonuria is influenced by various genetic factors. PMID- 11043162 TI - Mutation analysis anticipates dietary requirements in phenylketonuria. AB - Phenylalanine hydroxylase (PAH) deficiency is inherited as an autosomal recessive trait and the associated hyperphenylalaninaemia phenotype is highly variable, primarily due to a great allelic heterogeneity at the PAH locus (approximately 400 disease-associated mutations are known). The arbitrary classification of PAH deficiency on the basis of clinical parameters has been complicated by the lack of international guidelines, leading to a wide confusion in both methodology and terminology. Recently, significant improvements in methods for detection of mutations have paved the way for an alternative system for classification of PAH deficiency, which is based solely on PAH genotypes. This paper gives a summary of the recent progress made in establishing a direct correlation between individual PAH mutations and biochemical and metabolic phenotypes, including the use of "functionally hemizygous" patients to classify both common and rare mutant alleles, and a simple and general model to predict the combined phenotypic effect of two mutant PAH alleles. CONCLUSION: Genotype-based prediction of the biochemical phenotype is now feasible in the majority of newborns with hyperphenylalaninemia, which may be useful for refining diagnosis and anticipating dietary requirements. A recent observation suggests that the genotype also determines cognitive development if dietary therapy is discontinued at 6 years of age. PMID- 11043163 TI - PAH gene mutation analysis in clinical practice--comments on mutation analysis anticipates dietary requirements in phenylketonuria. PMID- 11043165 TI - [An outline of brain tumor classification]. PMID- 11043164 TI - The International Collaborative Study of Maternal Phenylketonuria: status report 1998. AB - The Maternal Phenylketonuria Study began in 1984 and during the intervening years, 572 pregnancies in hyperphenylalaninemic women and 99 controls and their outcomes have been evaluated. Among hyperphenylalaninemic women who delivered a live infant, only 15.9% were treated and in metabolic control preconceptually, however, another 18.4% were in control by 10 weeks. Compared to the results reported by Lenke and Levy in 1980, there is a marked improvement in outcome with treatment. Microcephaly was unusual in preconceptually treated pregnancies with well controlled phenylalanine restricted diets. Even in pregnancies that established control after conception but before the 8th week, congenital heart disease did not occur in the offspring, however, it did occur in 12% of pregnancies not achieving control until after 10 weeks of pregnancy. CONCLUSION: The recommended level of blood phenylalanine during pregnancy is 120-360 mumol/l. Best results were obtained by close cooperation between the attending obstetrician and a metabolic team experienced in the care of persons with phenylketonuria. PMID- 11043166 TI - [Astrocytoma]. PMID- 11043167 TI - [Anaplastic (malignant) astrocytoma]. PMID- 11043168 TI - [Glioblastoma multiforme]. PMID- 11043169 TI - [Pilocytic astrocytoma]. PMID- 11043170 TI - [Pleomorphic xanthoastrocytoma]. PMID- 11043171 TI - [Subependymal giant cell astrocytoma (tuberous sclerosis)]. PMID- 11043172 TI - [Oligodendroglioma]. PMID- 11043173 TI - [Anaplastic oligodendroglioma]. PMID- 11043174 TI - [Ependymoma]. PMID- 11043175 TI - [Anaplastic (malignant) ependymoma]. PMID- 11043176 TI - [Myxopapillary ependymoma]. PMID- 11043177 TI - [Subependymoma]. PMID- 11043178 TI - [Oligo-astrocytoma]. PMID- 11043179 TI - [Anaplastic (malignant) oligo-astrocytoma]. PMID- 11043180 TI - [Choroid plexus tumor--choroid plexus papilloma and choroid plexus carcinoma]. PMID- 11043181 TI - [Astroblastoma]. PMID- 11043182 TI - [Polar spongioblastoma]. PMID- 11043183 TI - [Gliomatosis cerebri]. PMID- 11043184 TI - [Gangliocytoma]. PMID- 11043185 TI - [Dysplastic gangliocytoma of cerebellum (Lhermitte-Duclos disease)]. PMID- 11043187 TI - [Dysembryoplastic neuroepithelial tumor (DNT)]. PMID- 11043186 TI - [Desmoplastic infantile ganglioglioma (DIG)]. PMID- 11043188 TI - [Ganglioglioma]. PMID- 11043189 TI - [Anaplastic ganglioglioma]. PMID- 11043190 TI - [Central neurocytoma]. PMID- 11043192 TI - [Olfactory neuroblastoma (esthesioneuroblastoma)]. PMID- 11043191 TI - [Paraganglioma of the filum terminale]. PMID- 11043193 TI - [Pineocytoma]. PMID- 11043194 TI - [Pineoblastoma]. PMID- 11043195 TI - [Mixed/transitional pineal tumor]. PMID- 11043196 TI - [Medulloepithelioma]. PMID- 11043197 TI - [Neuroblastoma]. PMID- 11043198 TI - [Ependymoblastoma]. PMID- 11043199 TI - [Primitive neuroectodermal tumours (PNETs)]. PMID- 11043200 TI - [Medulloblastoma]. PMID- 11043201 TI - [Schwannoma]. PMID- 11043202 TI - [Neurofibroma]. PMID- 11043204 TI - [Meningioma]. PMID- 11043203 TI - [Malignant peripheral nerve sheath tumor]. PMID- 11043205 TI - [Atypical meningioma]. PMID- 11043207 TI - [Anaplastic (malignant) meningioma]. PMID- 11043206 TI - [Papillary meningioma]. PMID- 11043208 TI - [Osteocartilaginous tumors]. PMID- 11043209 TI - [Lipoma]. PMID- 11043210 TI - [Fibrous histiocytoma]. PMID- 11043211 TI - [Hemangiopericytoma]. PMID- 11043212 TI - [Chondrosarcoma]. PMID- 11043213 TI - [Malignant fibrous histiocytoma]. PMID- 11043214 TI - [Rhabdomyosarcoma]. PMID- 11043215 TI - [Meningeal sarcomatosis]. PMID- 11043217 TI - [Hemangioblastoma]. PMID- 11043216 TI - [Primary melanocytic lesion]. PMID- 11043218 TI - [Primary central nervous system malignant lymphoma]. PMID- 11043219 TI - [Plasmacytoma]. PMID- 11043220 TI - [Granulocytic sarcoma (chloroma)]. PMID- 11043222 TI - [Embryonal carcinoma]. PMID- 11043221 TI - [Germinoma]. PMID- 11043223 TI - [Yolk sac tumor]. PMID- 11043224 TI - [Choriocarcinoma]. PMID- 11043225 TI - [Teratoma]. PMID- 11043226 TI - [Mixed germ cell tumors]. PMID- 11043227 TI - [Rathke's cleft cyst]. PMID- 11043228 TI - [Epidermoid cyst]. PMID- 11043229 TI - [Dermoid cyst]. PMID- 11043230 TI - [Colloid cyst]. PMID- 11043231 TI - [Enterogenous cyst]. PMID- 11043232 TI - [Neuroglial cyst]. PMID- 11043234 TI - [Hypothalamic neuronal hamartoma]. PMID- 11043233 TI - [Granular-cell tumor (choristoma, pituicytoma)]. PMID- 11043235 TI - [Nasal glial heterotopia]. PMID- 11043236 TI - [Plasma cell granuloma]. PMID- 11043237 TI - [Pituitary adenoma, pituitary carcinoma]. PMID- 11043238 TI - [Craniopharyngioma]. PMID- 11043239 TI - [Glomus jugulare tumor]. PMID- 11043240 TI - [Chordoma]. PMID- 11043241 TI - [Chondroma]. PMID- 11043242 TI - [Chondrosarcoma]. PMID- 11043243 TI - [Carcinoma]. PMID- 11043244 TI - [Adenoid cystic carcinoma (cylindroma)]. PMID- 11043245 TI - [Metastatic brain tumor]. PMID- 11043246 TI - [Brain tumors arising from frontal and parietal lobes]. PMID- 11043247 TI - [Brain tumor in the temporal lobe (including middle cranial fossa)]. PMID- 11043248 TI - [Tumors in the occipital lobe and related structures including tentorium cerebelli]. PMID- 11043249 TI - [Lateral ventricular tumors]. PMID- 11043250 TI - [Tumors in the pituitary and chiasmal region]. PMID- 11043251 TI - [Third ventricle tumor]. PMID- 11043253 TI - [Neoplasm of the brain stem region]. PMID- 11043252 TI - [Fourth ventricular tumors]. PMID- 11043254 TI - [Cerebellar tumor]. PMID- 11043255 TI - [Cerebello-pontine angle tumor]. PMID- 11043256 TI - [Tumors in the cavernous sinus (cavernous sinus tumor)]. PMID- 11043257 TI - [Brain tumors located in deep cortical regions]. PMID- 11043258 TI - [Pineal tumors]. PMID- 11043259 TI - [Posterior cranial fossa skull base tumors (clival, foramen magnum, and jugular foramen tumors)]. PMID- 11043260 TI - [Syndrome of orbit]. PMID- 11043261 TI - [Bone tumor of the skull]. PMID- 11043262 TI - [Spinal tumor, extradural tumor, intradural extramedullary and intramedullary tumor]. PMID- 11043263 TI - [Peripheral nerve tumors]. PMID- 11043264 TI - [Malformation: introduction and classification]. PMID- 11043265 TI - [Craniorachischisis totalis]. PMID- 11043266 TI - [Cranium bifidum, meningoencephalocele]. PMID- 11043267 TI - [Cephalocele]. PMID- 11043268 TI - [Anencephaly]. PMID- 11043269 TI - [Cranium bifidum occultum]. PMID- 11043270 TI - [Myeloschisis]. PMID- 11043272 TI - [Lipomeningocele]. PMID- 11043271 TI - [Spina bifida cystica]. PMID- 11043273 TI - [Diastematomyelia]. PMID- 11043274 TI - [Dimyelia, diplomyelia]. PMID- 11043275 TI - [Tethered cord syndrome]. PMID- 11043276 TI - [Congenital dermal sinus]. PMID- 11043277 TI - [Anterior dysrhaphic disturbance]. PMID- 11043278 TI - [Caudal regression syndrome]. PMID- 11043279 TI - [Spina bifida occulta]. PMID- 11043280 TI - [Atelencephaly]. PMID- 11043281 TI - [Holoprosencephaly]. PMID- 11043282 TI - [Septo-optic dysplasia]. PMID- 11043283 TI - [So-called diencephalic cyst and related diseases]. PMID- 11043284 TI - [Cerebral hemihypoplasia/aplasia]. PMID- 11043285 TI - [Hypoplasia/aplasia of the cerebellar hemisphere]. PMID- 11043286 TI - [Hypoplasia/aplasia of the cerebellar vermis including Joubert syndrome]. PMID- 11043287 TI - [Dysplasias of cerebellar cortex]. PMID- 11043288 TI - [Isolated lissencephaly sequence]. PMID- 11043289 TI - [Miller-Dieker syndrome]. PMID- 11043290 TI - [Focal agyria/pachygyria, partial lissencephaly]. PMID- 11043291 TI - [Congenital muscular dystrophy (Fukuyama type)]. PMID- 11043292 TI - [Walker-Warburg syndrome]. PMID- 11043294 TI - [Bilateral perisylvian polymicrogyria/bilateral perisylvian syndrome]. PMID- 11043293 TI - [Muscle-eye brain disease]. PMID- 11043295 TI - [Bilateral symmetric polymicrogyria]. PMID- 11043296 TI - [Asymmetric polymicrogyria]. PMID- 11043297 TI - [Schizencephaly]. PMID- 11043298 TI - [Periventricular nodular heterotopia (PNH)]. PMID- 11043299 TI - [Subcortical band heterotopia (SBH)]. PMID- 11043300 TI - [Leptomeningeal heterotopia]. PMID- 11043301 TI - [Focal heterotopia]. PMID- 11043302 TI - [Hemimegalencephaly]. PMID- 11043303 TI - [Neurofibromatosis type 1]. PMID- 11043304 TI - [Neurofibromatosis type 2 (NF2)]. PMID- 11043305 TI - [Tuberous sclerosis (TS)]. PMID- 11043306 TI - [Hypomelanosis of Ito]. PMID- 11043307 TI - [von Hippel-Lindau (VHL) disease]. PMID- 11043308 TI - [Ataxia-telangiectasia (Louis Bar syndrome)]. PMID- 11043309 TI - [Cockayne syndrome]. PMID- 11043310 TI - [Xeroderma pigmentosum]. PMID- 11043312 TI - [Sturge-Weber syndrome]. PMID- 11043311 TI - [Phacomatosis and tumors]. PMID- 11043313 TI - [Incontinentia pigmenti]. PMID- 11043314 TI - [Linear nevus sebaceus syndrome]. PMID- 11043315 TI - [Genetic megalencephaly]. PMID- 11043316 TI - [Megalencephaly with neurologic disorder]. PMID- 11043317 TI - [Megalencephaly with gigantism, cerebral gigantism (Sotos syndrome)]. PMID- 11043319 TI - [Scaphocephaly]. PMID- 11043318 TI - [Metabolic megalencephaly]. PMID- 11043320 TI - [Dolichocephaly]. PMID- 11043321 TI - [Brachycephaly]. PMID- 11043322 TI - [Oxycephaly, acrocephaly]. PMID- 11043323 TI - [Turricephaly]. PMID- 11043324 TI - [Plagiocephaly]. PMID- 11043325 TI - [Trigonocephaly]. PMID- 11043326 TI - [Crouzon craniofacial dysostosis]. PMID- 11043327 TI - [Apert syndrome]. PMID- 11043328 TI - [Carpenter syndrome (acrocephalopolysyndactyly type II)]. PMID- 11043329 TI - [Saethre-Chotzen syndrome]. PMID- 11043330 TI - [Pfeiffer syndrome]. PMID- 11043331 TI - [Syringomyelia]. PMID- 11043332 TI - [Chiari malformation]. PMID- 11043333 TI - [Dandy-Walker syndrome and Dandy-Walker variant]. PMID- 11043334 TI - [Klippel-Feil syndrome]. PMID- 11043335 TI - [Hypoplasia of the brain stem]. PMID- 11043336 TI - [Basilar impression]. PMID- 11043337 TI - [Agenesis of the corpus callosum]. PMID- 11043338 TI - [Subependymal pseudocyst]. PMID- 11043339 TI - [Agenesis of cranial nerve nuclei]. PMID- 11043340 TI - [Aqueductal stenosis]. PMID- 11043341 TI - [Congenital aqueductal stenosis]. PMID- 11043342 TI - [Colpocephaly]. PMID- 11043343 TI - [Porencephaly]. PMID- 11043345 TI - [Hydranencephaly]. PMID- 11043344 TI - [Multicystic encephalopathy]. PMID- 11043346 TI - [Ulegyria]. PMID- 11043347 TI - [Hypomyelination]. PMID- 11043348 TI - [Retarded myelination]. PMID- 11043349 TI - [Primary diseases affecting the gray matter]. PMID- 11043350 TI - [Primarily diseases affecting white matter/dysmyelination]. PMID- 11043351 TI - [Arachnoid cyst]. PMID- 11043352 TI - [Subdural hygroma (hydroma)]. PMID- 11043353 TI - [Choroid plexus cyst]. PMID- 11043354 TI - [Mega cisterna magna]. PMID- 11043355 TI - [Microcephalic syndrome]. PMID- 11043356 TI - [Syndrome accompanied with hydrocephalus]. PMID- 11043357 TI - [Encephaloclastic atelencephaly]. PMID- 11043358 TI - [Encephaloclastic hydranencephaly]. PMID- 11043359 TI - [Multicystic encephalomalacia]. PMID- 11043360 TI - [Encephaloclastic schizencephaly]. PMID- 11043361 TI - [Encephaloclastic porencephaly]. PMID- 11043362 TI - [Aqueductal stenosis]. PMID- 11043363 TI - [Destructive hydrocephalus]. PMID- 11043364 TI - [Damage of corpus callosum]. PMID- 11043365 TI - [Absence of septum pellucidum]. PMID- 11043366 TI - [Cerebral atrophy]. PMID- 11043367 TI - [Cerebellar atrophy]. PMID- 11043368 TI - [Acquired demyelinating diseases]. PMID- 11043369 TI - [Intracranial calcification]. PMID- 11043370 TI - [Hemorrhage in the choroid plexus]. PMID- 11043371 TI - [Subependymal hemorrhage]. PMID- 11043372 TI - [Other parenchymatous hemorrhage]. PMID- 11043373 TI - [Subdural hematoma]. PMID- 11043374 TI - [Subdural fluid collection]. PMID- 11043375 TI - [Periventricular leukomalacia]. PMID- 11043376 TI - [Congenital infarction]. PMID- 11043377 TI - [Non-decussation of pyramidal tract]. PMID- 11043378 TI - Endothelin and the "seventh inning stretch". PMID- 11043379 TI - The role of CD40 in peripheral T cell tolerance and immunity. AB - CD40 and CD40 ligand (CD40L) have been implicated as important molecules for the transformation of nonactivated antigen-presenting cells (APC) into cells that are potent inducers of cytotoxic T lymphocyte (CTL) immunity. The onset of a successful immune response lies within the control of the CD4+ T helper cells which, after specific antigen recognition, can up-regulate CD40L and subsequently activate APC through CD40 signaling. Triggering of CD40 with antibodies in vivo can replace the need for CD40L-expressing CD4+ T helper cells for cross-priming of CTL. Blocking of CD40-CD40L interactions can also have profound effects on the generation of T cell immunity. Interestingly, differential involvement of CD40/CD40L in immune responses can be observed between various immunological sites in the body. In most sites of the periphery interruption of CD40-CD40L interactions can lead to the induction of T cell tolerance whereas in mucosal tissues this interruption can lead to abrogation of T cell tolerance. Furthermore, in vivo CD40 activation can convert specific T cell tolerance following peptide vaccination into efficient T cell priming. Thus intervention of CD40-CD40L interactions can result in enhancement or down-modulation of T cell reactivity and therefore modulation of these interactions may form the foundation of new treatment modalities directed against malignancies, allergies, organ rejections and autoimmunity. PMID- 11043380 TI - Intron 4 polymorphism of the endothelial nitric oxide synthase gene is associated with elevated blood pressure in type 2 diabetic patients with coronary heart disease. AB - The endothelial nitric oxide synthase (eNOS) gene is responsible for constitutive nitric oxide synthesis and arterial vasodilatation. Recently two polymorphisms, the 27-bp repeat sequence in intron 4 and the Glu298Asp substitution in exon 7 of the eNOS gene have been reported to be related to coronary heart disease (CHD). We screened these polymorphisms of the eNOS gene in 308 unrelated nondiabetic subjects with CHD, in 251 unrelated patients with type 2 diabetes with CHD, and in 110 randomly selected healthy subjects without CHD. The 4a and Asp298 allele frequencies of the eNOS gene were 0.19 and 0.36 in nondiabetic patients with CHD, 0.21 and 0.27 in type 2 diabetic patients with CHD, and 0.16 and 0.31 in nondiabetic subjects without CHD (n.s. between the groups). The Asp298 allele in exon 7 of the eNOS gene was not associated with elevated blood pressure in any of the study groups. Among type 2 diabetic patients with CHD the 4a allele in intron 4 of the eNOS gene was associated with elevated levels of systolic (P=0.035) and mean arterial blood pressure (P=0.040). In nondiabetic subjects these associations were not statistically significant. When all study groups were pooled in statistical analysis the 4a allele of the eNOS gene was associated with elevated diastolic (P=0.032) and mean (P=0.030) arterial blood pressure even after adjustment for confounding factors. We conclude that the 4a allele of the eNOS gene is not associated with CHD or type 2 diabetes, but that it is related to elevated blood pressure levels particularly among type 2 diabetic patients with CHD. PMID- 11043381 TI - Induction of putative tumor-suppressing genes in Rat-1 fibroblasts by oncogenic Raf-1 as evidenced by robot-assisted complex hybridization. AB - The growth factor receptor-dependent protein kinase Raf-1 is activated by GTP bound Ras, thereby activating the mitogen-activated protein kinase pathway. To study the role of Raf in transformation we transduced Rat-1 cells with a tetracycline-regulatable retroviral vector encoding the constitutively active oncogenic C-terminal fragment of the human Raf-1 protein. Using subtractive hybridization of mRNAs from induced and noninduced cells and robot-assisted screening by complex hybridization, Raf-induced genes with various different characteristics of induction were investigated. Among the strongly induced genes were those involved in carcinogenesis such as metalloproteinases 3, 10 and 13, cathepsin L, ornithine decarboxylase, and putative tumor-suppressing genes such as monocyte chemoattracting protein 1, interferon-induced protein 10, a recently identified 2'-5' oligoadenylate synthetase-like protein, and plasminogen activator inhibitor type 2. Other components of the plasminogen activator system were not induced. Plasminogen activator inhibitor type 2 is a down-regulator of the proteolytic cascade consisting of various metalloproteinases, some of which are induced by a carboxy-terminal Raf mutant (RafCT). In conclusion, RafCT induces factors which act in a conflicting manner in respect of carcinogenesis, especially within the proteolytic system of the extracellular matrix. PMID- 11043383 TI - The state of the art microdissection and its downstream applications. PMID- 11043382 TI - Molecular cloning and expression of a novel klotho-related protein. AB - Klotho protein is a novel beta-glucosidase-like protein produced predominantly in the kidney. The klotho mouse, which genetically lacks klotho gene expression, manifests various systemic phenotypes resembling aging. In the present study we succeeded in isolating a novel human protein structurally related to klotho protein. The protein possesses one beta-glucosidase-like domain and is 42% identical with klotho protein at the amino acid level. Unlike klotho protein, it possesses neither a signal sequence nor a transmembrane domain, suggesting that it is a cytosolic protein, and thus was termed cytosolic beta-glucosidase-like protein-1 (cBGL1). By Northern blot analysis cBGL1 mRNA was expressed most abundantly in the liver, followed by the small intestine, colon, spleen, and kidney. When klotho and cBGL1 gene expression was examined in renal cell carcinoma tissues, both klotho and cBGL1 mRNA levels in tumors were lower than those in nontumor regions, suggesting that renal epithelial cells may lose klotho and cBGL1 gene expression during the course of malignant transformation. In conclusion, we describe the primary structure and gene expression of a novel protein related to klotho protein. PMID- 11043384 TI - The force of focused light in different areas of cell and molecular biology. PMID- 11043385 TI - Use of SMART-generated cDNA for differential gene expression studies. AB - A major limitation of gene expression profiling using microarrays has been the substantial amount of RNA required for standard probe labeling techniques, especially when working with clinical samples such as biopsies, microdissected tumors, and laser-captured cells. CLONTECH's PCR-based SMART technology (Switch Mechanism At the 5'end of RNA Templates) bypasses this problem by allowing accurate cDNA amplification from nanogram quantities of total RNA. This amount of RNA ensures that SMART amplification yields a pool of cDNA, which reflects the sample's original complexity and relative abundance of the original RNA sample. PMID- 11043386 TI - Microdissection of frozen sections. PMID- 11043387 TI - The LEICA microdissection system: design and applications. PMID- 11043388 TI - Quantitative gene expression analysis in microdissected archival tissue by real time RT-PCR. AB - Laser-assisted microdissection is a powerful tool for the analysis of morphologically defined cell populations. We report here that the combination of microdissection and real-time RT-PCR technologies together with an optimized RNA microscale extraction procedure allows the reproducible determination of gene expression levels in archival formalin-fixed, paraffin-embedded tissue samples. PMID- 11043389 TI - The ProteinChip System from Ciphergen: a new technique for rapid, micro-scale protein biology. PMID- 11043390 TI - Presence of genetic alterations in microdissected stroma of human colon and breast cancers. AB - We show by laser-assisted microdissection that frequent genetic alterations in non-hereditary invasive human colon and breast cancers (loss of heterozygosity and TP53 mutations) occur not only in the neoplastic epithelial cells, but also in the adjacent fibroblastic tumor stroma and that both components can share clonal features. Tumor cell-mesenchyme transitions are among the possible explanations for these findings. PMID- 11043391 TI - Microdissection and breast cancer. PMID- 11043392 TI - Positron-emission tomography with [18F]fluorodeoxyglucose. Part I. Biochemical uptake mechanism and its implication for clinical studies. AB - Over the past decades, Positron Emission Tomography has opened a new field of imaging. Nowadays, this technique is being used for diagnosing, staging disease as well as for prognostic stratification and monitoring therapy. In this respect, [18F]fluorodeoxyglucose (FdGlc) is by far the most commonly used PET agent. Many factors have been identified being responsible for a high uptake of this agent in malignancy. However, additional factors such as tumour treatment may interfere with the uptake mechanism. Knowledge of all these factors is a prerequisite for an optimal interpretation of PET studies and, consequently, for a reliable judgement of tumour status. In this article, a review is given of the factors influencing FdGlc uptake and the implications for clinical studies. PMID- 11043393 TI - Positron emission tomography with 2-[18F]fluoro-2-deoxy-D-glucose in oncology. Part II. The clinical value in detecting and staging primary tumours. AB - The tumoral uptake of 2-[18F]fluoro-2-deoxy-D-glucose (18FdGlc) is based upon enhanced glycolysis. Positron-emission tomography (PET) using 18FdGlc provides the physiological and metabolic information. 18FdGlc PET has been used successfully for assessing primary tumours and metastases, prognosis, and planning and for monitoring tumour therapy as well as for early detection of recurrent tumour growth. This review summarises the uptake mechanism of 18FdGlc in benign and malignant lesions, its relation to histopathology, and its clinical value for detecting and staging primary tumours. PMID- 11043394 TI - Epidemiology and biology of human urinary bladder cancer. AB - The bladder constitutes the most frequent localization of malignant tumors in the urinary tract. Further prognostic factors are molecular and cytogenetic alterations, which have been identified as key mechanisms in the carcinogenetic pathway of bladder cancer. Structural or numerical chromosomal alterations lead to the activation of a variety of cancer-inducing oncogenes as well as to the inactivation of various distinct antiproliferative tumor-suppressor genes. With regard to the biological heterogeneity in transitional cell carcinoma, which is also reflected in epidemiological data, the differing clinical course and the limited value of established prognosticators, the analysis of new molecular parameters has become of interest in predicting the prognosis of bladder cancer patients. In addition, the definition of high-risk patient groups that are at at risk of progression and recurrence is a further objective of urological research in this field. PMID- 11043395 TI - Biological effects of ionizing radiation on human blood compounds ex vivo. AB - PURPOSE: Blood compounds are irradiated ex vivo to prevent transfusion-associated graft-versus-host-disease. Recently, ex vivo irradiation of re-transfused wound blood has been proposed to prevent metastatic spread in patients with malignant tumors, an issue requiring different dose concepts. To determine effects on blood cells we examined the impact of various doses of ionizing radiation. METHODS: Full blood was irradiated with doses of 10-150 Gy. Potassium, LDH and hemoglobin levels were determined 2 h-96 h after irradiation. The lymphocyte proliferation after irradiation was measured by means of a lymphocyte-transformation assay. The impact of irradiation on mitogen-induced secretion of cytokines was determined by the ELISA technique, and P-selectin expression as an indicator of platelet activation was analyzed by flow-cytometry. RESULTS: Potassium levels increase with aging and irradiation dose. Mitogenic capacity is reduced by over 90% with moderate doses of 10-20 Gy, but a residual proliferation is still detectable up to 50 Gy. No enhancement of extracellular cytokine levels is detectable, but the cytokine release is reduced by radiation. Neither induction of platelet activation nor abrogation of activation has been detected. CONCLUSIONS: Doses of 30-50 Gy abrogate lymphocyte proliferation almost completely. In this range we did not observe severe adverse effects on blood transfusions. Hemolysis might be enhanced when the samples are stored for a longer period after irradiation. PMID- 11043396 TI - Study of immortalization and malignant transformation of human embryonic esophageal epithelial cells induced by HPV18 E6E7. AB - In order to study the effect of viruses and tumor promoters on the tumorigenicity of the esophagus, human embryonic esophageal epithelial cells were infected with human papilloma virus HPV18 E6E7-AAV in synergy with 12-O-tetradecanoylphorbol 13 acetate (TPA) to observe their malignant transformation. The cultured esophageal epithelial cells incubated with HPV18 E6E7-AAV were divided into two groups: the SHEEC1 group was exposed to TPA (5 ng/ml) for 4 weeks at the 5th passage of the cells; the SHEE group served as the control and was cultured in the same medium without TPA. The morphological phenotype, the DNA content during the cell cycle and the chromosomes were analyzed. The tumorigenicity was assessed by colony formation after cultivation in soft agar and transplanting the cells into nude mice. HPV18 E6E7 DNA was assayed by fluorescent in situ hybridization (FISH) and the polymerase chain reaction (PCR). The SHEE group, at its 20th passage, grew as a monolayer with the cells showing anchorage dependence and contact inhibition. The chromosome analysis showed diploidy, and soft-agar cultivation and injection into nude mice showed the cells to be non-tumorigenic. They were therefore immortalized cells. In contrast, the SHEEC1 group (TPA group) showed increased DNA synthesis and a proliferative index that was higher (45%) than that of the SHEE group (34%). The number of large colonies of dense multilayer cells (positively transformed foci) in soft agar was high in SHEEC1 group (4.0%) but low in the SHEE group (0.1%). Tumors resulting from transplantation were observed in all six nude mice injected subcutaneously with cells of the SHEEC1 group but no tumor developed in mice receiving cells of the SHEE group. In both groups of cells, HPV18 E6E7 DNA was positively detected by FISH and PCR. The malignant transformation of human embryonic epithelial cells was induced in vitro by HPV18 E6E7 in synergy with TPA. This is a good evidence for the close relationship between HPV and the etiology and pathogenicity of esophageal carcinoma. It is also a reliable model for studying the cellular and molecular mechanisms of carcinogenesis of esophageal carcinoma. PMID- 11043397 TI - Inhibitory effects of synthetic beta peptide on invasion and metastasis of liver cancer. AB - PURPOSE: To study the inhibitory effects of synthetic beta peptide on invasion and metastasis of liver cancer. METHODS: Membrane-type intercellular adhesion molecule-1 (ICAM-1) expression of SMMC-7721 cultured hepatoma cells (7721 cells) was detected by immunofluorescence cell flowmeter. The adhesion of 7721 cells to fibronectin (FN) was assayed by the MTT method. The adhesion of 7721 cells to 7721 cells, 7721 cells to endothelial cells, and 7721 cells to lymphocyte cells was detected by adhesion assay. LCI-D20 human liver cancer metastasis model in nude mice was used in this experiment. One hundred micrograms of beta peptide per mouse were injected subcutaneously after tumor was resected premetastatically or postmetastatically to observe its effect on liver cancer metastasis after hepatectomy. RESULTS: Membrane-type ICAM-1 expression of SMMC-7721 cells treated by beta peptide was lower than that of the untreated cells. The adhesion of 7721 cells to FN, 7721 cells to 7721 cells, 7721 cells to endothelial cells, and 7721 cells to lymphocyte cells was also lower in the beta peptide group than in the untreated group. CONCLUSIONS: beta Peptide can block the adhesion of 7721 cells to FN, 7721 cells to some host cells in vitro, and inhibit HCC metastasis of LCI D20 model posthepatectomy in vivo, so it could potentially act as an antimetastasis drug. PMID- 11043398 TI - Sonodynamically induced antitumor effect of Photofrin II on colon 26 carcinoma. AB - The sonodynamically induced antitumor effect of Photofrin II (PF), was evaluated in mice bearing colon 26 carcinoma. In order to find the optimum timing for ultrasonic exposure after the administration of PF, the PF concentrations in the plasma, skin, muscle, and tumor were measured. The antitumor effect was estimated by measuring the tumor size. Since the highest concentration of PF in the tumor was observed 24 h after administration, an ultrasonic exposure timing of 24 h after the intravenous administration of PF was chosen. When used alone, ultrasound showed a slight antitumor effect, which became increasingly significant as the dose of PF was increased, while use of PF alone showed no significant effect. From these results, it is concluded that PF significantly sensitizes solid tumors to ultrasound, demonstrating a synergistic antitumor effect. PMID- 11043399 TI - Analysis of cadherin/catenin complexes in transformed thyroid epithelial cells: modulation by beta 1 integrin subunit. AB - We have analysed the expression of cadherin/catenin complex molecules in PC C13 rat thyroid cells transformed in vitro with different oncogenes. No significant downregulation of either E-cadherin, alpha-, beta- and gamma-catenin was detected following the introduction of activated forms of myc, adenovirus E1A, ras, raf, myc + ras, E1A + raf. However, ras- and raf-transformed PC C13 cells showed altered adherens junctions. An altered distribution of cadherin/catenin complexes characterized by radially oriented membrane spikes perpendicular to cell edges was the most prominent feature evidenced by immunofluorescence. No beta1 integrin localization was observed in areas where this altered pattern of E-cadherin expression was detected. However, beta1 integrin subunit expression was detected at areas of cell-cell contact where E-cadherin showed a normal pattern of expression. Furthermore, ras- and raf-transformed PC C13 cells showed the ability to migrate in collagen gels, in contrast to their normal untransformed counterpart. Overexpression of beta1 integrin was found to restore normal E cadherin localization at cell-cell contacts and to partially inhibit the ability to migrate in collagen gels. Finally, two cell lines obtained by ras transformation in vivo, and derived from a rat primary thyroid carcinoma (TK6) and its lung metastasis (MPTK6), were found to have lost gamma-catenin expression. TK6 lost also E-cadherin expression and membrane localization of alpha-catenin. These results suggest that: i) in vitro thyroid cell transformation is associated to a change in cadherin/catenin complexes distribution rather than to a decrease in expression; ii) in vivo transformation is associated to the loss of expression of some of these molecules likely due to tumor progression; iii) alterations in beta1 integrin subunit expression can result in changes in cadherin/catenin function thus implying that an integrin cadherin synergy may exist in thyroid cells. PMID- 11043400 TI - Growth factor-induced morphological, physiological and molecular characteristics in cerebral endothelial cells. AB - The capacity of vascular endothelial cells to modulate their phenotype in response to changes in environmental conditions is one of the most important characteristics of this cell type. Since different growth factors may play an important signalling role in this adaptive process we have investigated the effect of endothelial cell growth factor (ECGF) on morphological, physiological and molecular characteristics of cerebral endothelial cells (CECs). CECs grown in the presence of ECGF and its cofactor heparin exhibit an epithelial-like morphology (type I CECs). Upon removal of growth factors, CECs develop an elongated spindle-like shape (type II CECs) which is accompanied by the reorganization of actin filaments and the induction of alpha-actin expression. Since one of the most important functions of CECs is the creation of a selective diffusion barrier between the blood and the central nervous system (CNS), we have studied the expression of junction-related proteins in both cell types. We have found that removal of growth factors from endothelial cultures leads to the downregulation of cadherin and occludin protein levels. The loss of junctional proteins was accompanied by a significant increase in the migratory activity and an altered protease activity profile of the cells. TGF-beta1 suppressed endothelial migration in all experiments. Our data provide evidence to suggest that particular endothelial functions are largely controlled by the presence of growth factors. The differences in adhesiveness and migration may play a role in important physiological and pathological processes of endothelial cells such as vasculogenesis or tumor progression. PMID- 11043401 TI - The effects of high magnitude cyclic tensile load on cartilage matrix metabolism in cultured chondrocytes. AB - Excessive mechanical load is thought to be responsible for the onset of osteoarthrosis (OA), but the mechanisms of cartilage destruction caused by mechanical loads remain unknown. In this study we applied a high magnitude cyclic tensile load to cultured chondrocytes using a Flexercell strain unit, which produces a change in cell morphology from a polygonal to spindle-like shape, and examined the protein level of cartilage matrixes and the gene expression of matrix metalloproteinases (MMPs), tissue inhibitors of matrix metalloproteinases (TIMPs) and proinflammatory cytokines such as IL-1beta and TNF-alpha. Toluidine blue staining, type II collagen immunostaining, and an assay of the incorporation of [35S]sulfate into proteoglycans revealed a decrease in the level of cartilage specific matrixes in chondrocyte cultures subjected to high magnitude cyclic tensile load. PCR-Southern blot analysis showed that the high magnitude cyclic tensile load increased the mRNA level of MMP-1, MMP-3, MMP-9, IL-1beta, TNF-alpha and TIMP-1 in the cultured chondrocytes, while the mRNA level of MMP-2 and TIMP-2 was unchanged. Moreover, the induction of MMP-1, MMP-3 and MMP-9 mRNA expression was observed in the presence of cycloheximide, an inhibitor of protein synthesis. These findings suggest that excessive mechanical load directly changes the metabolism of cartilage by reducing the matrix components and causing a quantitative imbalance between MMPs and TIMPs. PMID- 11043402 TI - Stage-specific apoptotic patterns during Drosophila oogenesis. AB - In the present study we demonstrate the existence of two apoptotic patterns in Drosophila nurse cells during oogenesis. One is developmentally regulated and normally occurs at stage 12 and the other is stage-specific and is sporadically observed at stages 7 and 8 of abnormally developed follicles. The apoptotic manifestation of the first pattern begins at stage 11 and is marked by a perinuclear rearrangement of the actin cytoskeleton and the development of extensive lobes and engulfments of the nurse cell nuclei located proximal to the oocyte. Consequently, at late stage 12 (12C), half of the nurse cell nuclei exhibit condensed chromatin, while at late stage 13 all the nuclei have fragmented DNA, as it is clearly shown by TUNEL assay. Finally, the apoptotic vesicles that are formed during stage 13, are phagocytosed by the neighboring follicle cells and at stage 14 the nurse cell nuclear remnants can be easily detected within the adjacent follicle cell phagosomes. In the second sporadic apoptotic pattern, all the nurse cell nuclei are highly condensed with fragmented DNA, accompanied by a completely disorganized actin cytoskeleton. When we induced apoptosis in Drosophila follicles through an etoposide and staurosporine in vitro treatment, we observed a similar pattern of stage-specific cell death at stages 7 and 8. These observations suggest a possible protective mechanism throughout Drosophila oogenesis that results in apoptosis of abnormal, damaged or spontaneously mutated follicles before they reach maturity. PMID- 11043403 TI - The receptor tyrosine phosphatase-like protein ICA512 binds the PDZ domains of beta2-syntrophin and nNOS in pancreatic beta-cells. AB - Islet cell autoantigen (ICA) 512 of type I diabetes is a receptor tyrosine phosphatase-like protein associated with the secretory granules of neurons and endocrine cells including insulin-secreting beta-cells of the pancreas. Here we show that in a yeast two-hybrid assay its cytoplasmic domain binds beta2 syntrophin, a modular adapter which in muscle cells interacts with members of the dystrophin family including utrophin, as well as the signaling molecule neuronal nitric oxide synthase (nNOS). The cDNA isolated by two-hybrid screening corresponded to a novel beta2-syntrophin isoform with a predicted molecular mass of 28 kDa. This isoform included the PDZ domain, but not the C-terminal region, which in full-length beta2-syntrophin is responsible for binding dystrophin related proteins. In vitro binding of the beta2-syntrophin PDZ domain to ICA512 required both ICA512's C-terminal region and an internal polypeptide preceding its tyrosine phosphatase-like domain. Immunomicroscopy and co immunoprecipitations from insulinoma INS-1 cells confirmed the occurrence of ICA512-beta2-syntrophin complexes in vivo. ICA512 also interacted in vitro with the PDZ domain of nNOS and ICA512-nNOS complexes were co-immunoprecipitated from INS-1 cells. Finally, we show that INS-1 cells, like muscle cells, contain beta2 syntrophin-utrophin oligomers. Thus, we propose that ICA512, through beta2 syntrophin and nNOS, links secretory granules with the actin cytoskeleton and signaling pathways involving nitric oxide. PMID- 11043404 TI - Peptide secretion in the cutaneous glands of South American tree frog Phyllomedusa bicolor: an ultrastructural study. AB - The development of the dermal glands of the arboreal frog Phyllomedusa bicolor was investigated by immunocytochemistry and electron microscopy. The 3 types of glands (mucous, lipid and serous) differed in size and secretory activity. The mucous and serous glands were apparent in the tadpole skin, whereas the lipid glands developed later in ontogenesis. The peptide antibiotics dermaseptins and the D-amino acid-containing peptide opioids dermorphins and deltorphins are abundant in the skin secretions of P. bicolor. Although these peptides differ in their structure and activity they are derived from precursors that have very similar preproregions. We used an antibody to the common preproregion of preprodermaseptins and preprodeltorphins and immunofluorescence analysis to show that only the serous glands are specifically involved in the biosynthesis and secretion of dermaseptins and deltorphins. Scanning and transmission electron microscopy revealed that the serous glands of P bicolor have morphological features, especially the secretory granules, which differ from those of the glands in Xenopus laevis skin. PMID- 11043405 TI - Sub-second quenched-flow/X-ray microanalysis shows rapid Ca2+ mobilization from cortical stores paralleled by Ca2+ influx during synchronous exocytosis in Paramecium cells. AB - Though only actual local free Ca2+ concentrations, [Ca2+], rather than total Ca concentrations, [Ca], govern cellular responses, analysis of total calcium fluxes would be important to fully understand the very complex Ca2+ dynamics during cell stimulation. Using Paramecium cells we analyzed Ca2+ mobilization from cortical stores during synchronous (< or = 80 ms) exocytosis stimulation, by quenched flow/cryofixation, freeze-substitution (modified for Ca retention) and X-ray microanalysis which registers total calcium concentrations, [Ca]. When the extracellular free calcium concentration, [Ca2+]e, is adjusted to approximately 30 nM, i.e. slightly below the normal free intracellular calcium concentration, [Ca2+]i = 65 nM, exocytosis stimulation causes release of 52% of calcium from stores within 80 ms. At higher extracellular calcium concentration, [Ca2+]e = 500 microM, Ca2+ release is counterbalanced by influx into stores within the first 80 ms, followed by decline of total calcium, [Ca], in stores to 21% of basal values within 1 s. This includes the time required for endocytosis coupling (350 ms), another Ca2+-dependent process. To confirm that Ca2+ mobilization from stores is superimposed by rapid Ca2+ influx and/or uptake into stores, we substituted Sr2+ for Ca2+ in the medium for 500 ms, followed by 80 ms stimulation. This reveals reduced Ca signals, but strong Sr signals in stores. During stimulation, Ca2+ is spilled over preformed exocytosis sites, particularly with increasing extracellular free calcium, [Ca2+]e. Cortically enriched mitochondria rapidly gain Ca signals during stimulation. Balance calculations indicate that total Ca2+ flux largely exceeds values of intracellular free calcium concentrations locally required for exocytosis (as determined previously). Our approach and some of our findings appear relevant also for some other secretory systems. PMID- 11043406 TI - Reporter-linked monitoring of transgene expression in living cells using the ecdysone-inducible promoter system. AB - Inducible promoter systems such as the ecdysone-inducible system or the tetracycline-regulated expression systems have proven to be powerful tools in studying gene function. In practice, such systems have met with the difficulty that either the vector expressing the transactivator gene or the vector carrying the response element are frequently silenced by flanking genomic sequences after stable integration. In order to identify those cells in a heterogeneous population in which a transgene is expressed from an ecdysone-inducible promoter, we have created the vector p2ER-EGFP/mcs that contains two ecdysone-inducible expression cassettes in tandem. Using two reporter genes, lacZ and green fluorescent protein (EGFP), we demonstrate that the expression of both genes can be co-induced from a very low baseline in CHO cells expressing the modified ecdysone receptor and the retinoid X receptor. The expression of EGFP and lacZ from vector p2ER-EGFP/lacZ follows the same Muristerone A concentration dependence as that of EGFP from vector pER-EGFP, indicating that the juxtaposition of the two inducible promoters in vector p2ER-EGFP/mcs does not cause cross interference between them. We suggest that this modification of the ecdysone-inducible promoter system will allow for the visual control of the induced expression of other genes by Muristerone A. PMID- 11043407 TI - Ca2+ oscillations and the cell cycle at fertilisation of mammalian and ascidian eggs. AB - At fertilisation of mammalian and ascidian eggs the sperm induces a series of Ca2+ oscillations. These Ca2+ oscillations are triggered by a sperm-borne Ca2+ releasing factor whose identity is still unresolved. In both mammals and ascidians Ca2+ oscillations in eggs are associated with the period leading up to exit from meiosis and entry into the first embryonic cell cycle. Thus, in mammals Ca2+ oscillations continue for several hours but are complete by within 30 min in the ascidian. In mammals and ascidians Ca2+ oscillations stop at around the time when pronuclei form in the 1-cell embryo. There is evidence to show that cell cycle factors are important in regulating the fertilisation Ca2+ signal. If the formation of pronuclei is blocked either in mammals (by spindle disruption) or in ascidians (by clamping maturation promoting factor levels high) then Ca2+ oscillations continue indefinitely. Here, we explore the nature of the sperm Ca2+ releasing factor and examine the relationship between cell cycle resumption and the control of Ca2+ oscillations at fertilisation. PMID- 11043408 TI - NAADP-induced calcium release in sea urchin eggs. AB - Nicotinic acid adenine dinucleotide phosphate (NAADP) is the most potent activator of Ca2+ release from intracellular stores described. It acts on a mechanism distinct from inositol trisphosphate and ryanodine receptors, the two major Ca2+ release channels characterised. NAADP-gated Ca2+ release channels do not appear to be regulated by Ca2+ and may be better suited for triggering Ca2+ signals rather than propagating them. They exhibit a remarkable pharmacology for a putative intracellular Ca2+ release channel in that they are selectively blocked by potassium and L-type Ca2+ channel antagonists. Furthermore, in contrast to microsomal Ca2+ stores expressing IP3Rs and RyRs, those sensitive to NAADP are thapsigargin-insensitive, suggesting that they may be expressed on a different part of the endoplasmic reticulum. Perhaps the most unusual feature of the NAADP-gated Ca2+ release mechanisms is its inactivation properties. Unlike the mechanisms regulated by IP3 and cADPR in sea urchin eggs which after induction of Ca2+ release appear to become refractory to subsequent activation, very low concentrations of NAADP are able to inactivate NAADP-induced Ca2+ release fully at concentrations well below those required to activate Ca2+ release. The mechanism and physiological significance of this most unusual desensitisation phenomenon are unclear. More recently, NAADP has been shown to mobilise Ca2+ in ascidian oocytes, brain microsomes and pancreatic acinar cells suggesting a more widespread role in Ca2+ signalling. A possible role for this novel Ca2+ release mechanism in sea urchin egg fertilisation is discussed. PMID- 11043409 TI - The initiation and propagation of the fertilization wave in sea urchin eggs. AB - Calcium waves sweep across most eggs of the deuterostome lineage at fertilization. The precise timing of the initiation and propagation of a fertilization calcium wave has been best studied in sea urchin embryos, since the rapid depolarization caused by sperm egg fusion can be detected as a calcium influx using confocal imaging of calcium indicator dyes. The time between sperm egg fusion and the first sign of the calcium increase that constitutes the calcium wave is comparable to the time it takes for the wave to sweep across the egg, once initiated. The latency and rise time of the calcium response is sensitive to inhibitors of the InsP3 signalling pathway, as reported previously. Using calcium green dextran and confocal microscopy, we confirm that the propagation time of the calcium wave is lengthened and that initiation of the calcium wave involves activation of calcium release at hot spots that may represent clusters of calcium release channels, as has been seen in other cell types. PMID- 11043410 TI - Egg activation: upstream of the fertilization calcium signal. AB - Interaction of sperm and egg at fertilization induces well-coordinated molecular events including specific recognition between species, adhesion and fusion, that lead to the formation of a zygote, a totipotent cell that develops into a new individual. A calcium signal, common to a great number of species, from marine invertebrates to mammals, is essential to activate the metabolism of the unfertilized oocyte. However, how fertilization triggers this calcium signal and initiates development of the early embryo is far from understood. The signaling pathways activated in eggs may be similar to those described in somatic cells, since changes in intracellular free calcium and in mitosis activating protein (MAP) kinase activity occur in both systems after activation. Several hypotheses are currently proposed, implying a spermatic ligand binding to a specific receptor expressed at the egg surface, or where the fused sperm either allows the transit of external calcium into the egg or injects one (or several) activating factor(s). It is still not known which of these ideas is true. We concentrate in this review on the possible signaling pathways involving IP3 (inositol trisphosphate), since its production is involved in most species to generate the fertilization calcium wave. PMID- 11043411 TI - Adhesion proteins expressed on human gamete surfaces and egg activation. AB - The initiation and propagation of a Ca2+ signal through the egg seems to be the pivotal event in triggering of meiosis resumption. Over the past decade evidence has accumulated suggesting that sperm contact is essential for this phenomenon to occur in most physiological groups. Given their ability to transduce signals, adhesive proteins which are involved in various binding mechanisms such as cell migration, lymphocyte activation, phagocytosis and virus fusion may play a similar role in fertilization. They have been the subject of serious investigation in non-human mammals and some emerging data indicate that they are active in humans as well. Our goal is to review the presence of such molecules on human gametes and their relevant physiological role, i.e., integrins and their ligands, selectins, IgG Fc receptors and leucocyte differentiation markers. We will discuss how they might trigger egg activation through signaling pathways in light of their identified functions in other adhesion systems. The putative participation of specific human sperm proteins will also be evaluated. PMID- 11043412 TI - Localised MPF regulation in eggs. AB - In this review we discuss the evidence that activation and inactivation of M phase promoting factor (MPF), the universal mitotic activator, are regulated locally within the cell, and consider the mechanisms that might be responsible. Localised initiation of MPF activation has been demonstrated in Xenopus eggs and egg fragments by examination of the timing of surface contraction waves (SCWs), indicators of MPF activity, and confirmed by direct measurement of MPF in such fragments. Both the timing and the site of SCW initiation relate to the presence of nuclei and of associated centriole-nucleated microtubules. Localised MPF activation is likely to occur in the perinuclear cytoplasm as well as within the nucleus. Studies in a number of cell types show that the perinuclear/centrosomal region is the site of accumulation of MPF itself (the cyclin B-Cdc2 kinase complex) and of many of its molecular regulators. It also harbours calcium regulating machinery, and in sea urchin eggs is the site of transient calcium release at the onset of mitosis. During mitosis MPF, regulatory molecules and calcium signalling components associate with spindle structures. Inactivation of MPF to end mitosis has been shown to be initiated locally at the mitoic spindle in Drosophila embryos. In sea urchin and frog eggs, calcium transients are required for both mitotic entry and exit and in mouse eggs, MPF inactivation requires both a calcium signal and an intact spindle. It thus appears that calcium signals coinciding with localised accumulation of MPF regulators are required first to set off and/or amplify the MPF activation process around the nucleus, and later to promote MPF inactivation via cyclin B destruction. Calcium release from sequestering machinery organised around nuclear and astral structures may act co-operatively with localised MPF regulatory molecules to trigger both mitotic entry and exit. PMID- 11043413 TI - The acrosome reaction in human spermatozoa. AB - During gamete interaction, sperm acrosome reaction (AR) induced by oocyte investment is a prerequisite event for the spermatozoa to pass through the zona pellucida (ZP), fuse with and penetrate the oocyte. Progesterone (P4), secreted by cumulus cells, is an important cofactor for the occurrence of this exocytosis event. The AR results from the fusion between outer acrosomal and plasma membranes, leading to inner acrosomal membrane exposure. Binding of agonists, P4 or ZP3 glycoprotein, to plasma membrane sperm receptors activates intraspermatic signals and enzymatic pathways involved in the AR. Among the proteins or glycoproteins described as potential sperm receptors for ZP, Gi/Go protein coupled and tyrosine kinase receptors have been described. Sperm receptors for P4 are poorly characterized, except a putative GABA(A)-like receptor. ZP- and P4 promoted AR is mediated by an obligatory intracellular calcium increase, appearing first at the acrosome equatorial segment and spreading throughout the head. The plasma membrane channels involved in calcium entry are operated by a plasma membrane depolarization and protein phosphorylations mediated by protein kinase C and tyrosine kinase protein. Part of the calcium increase could also be due to intracellular store release through IP3- and nucleotide (cAMP)-gated channels. Besides adenylate cyclase and phospholipase C activations, intracellular calcium increase also stimulates PLA2 activity and actin depolymerization, leading to membrane fusion. Evaluation of AR by staining or fluorescent probes can be useful to predict fertilization success and to direct the therapeutic strategy in male infertility. PMID- 11043414 TI - The soluble mammalian sperm factor protein that triggers Ca2+ oscillations in eggs: evidence for expression of mRNA(s) coding for sperm factor protein(s) in spermatogenic cells. AB - At fertilisation in mammals the sperm initiates a series of Ca2+ oscillations that activate development. One theory of signalling at fertilisation suggests that the sperm contains a soluble protein factor that causes these Ca2+ oscillations by entering the egg after sperm-egg membrane fusion. This theory is supported by the finding that, in some species, injection of sperm protein extracts into eggs triggers a pattern of Ca2+ oscillations similar to those seen at fertilisation. So far, all the direct evidence for a sperm factor has been based upon the injection of soluble proteins from mature sperm. Here, we demonstrate that injection of mRNA extracted from hamster spermatogenic cells also leads to generation of prolonged Ca2+ oscillations in mouse eggs. The ability of spermatogenic cell mRNA to induce Ca2+ oscillations is dependent upon translation into protein and also appears to be specific to spermatogenic cells since injection of mRNA isolated from somatic tissues into eggs was ineffective. These data support the hypothesis that sperm contain a soluble, cytosolic protein factor that induces Ca2+ oscillations in eggs at fertilisation. These data are discussed in the light of our recent findings that suggest that the sperm factor possesses a phospholipase C activity. PMID- 11043415 TI - Bilateral asymmetry of the inositol trisphosphate-mediated calcium signaling in two-cell ascidian embryos. AB - In ascidian oocytes, numerous calcium signaling events occur at fertilization which contribute to resume and complete meiosis, and determine the three embryonic axes. The main ooplasmic and intracellular calcium channels at work in the calcium signaling of the one-cell embryo have different roles and fates when the first mitosis begins. By whole-cell patch-clamp recording, we observed different families of these calcium channels in the blastomeres of Phallusia mammillata two-cell ascidian embryos. Membrane capacitance has been measured to evaluate the oocyte and blastomere surface area, allowing certification of the exact time of cell division. At the two-cell stage, no difference was observed in the density of voltage-dependent calcium channels in each blastomere, or in the ryanodine-sensitive calcium stores. In contrast, a bilateral asymmetry was recorded for the ooplasmic channels responsible for calcium entry after calcium store depletion: they could be activated only in the blastomere not wearing the polar bodies. The same laterality was observed in the InsP3-induced internal calcium release. Moreover, this asymmetry included a one-way communication in the InsP3-dependent calcium signaling between the two blastomeres. These results enhance the understanding of the early steps of development, and underscore the interest for ascidians in studies of polarity patterning. PMID- 11043416 TI - Role of dihydropyridine-sensitive calcium channels in meiosis and fertilization in the bivalve molluscs Ruditapes philippinarum and Crassostrea gigas. AB - Prophase-arrested oocytes of Ruditapes philippinarum can not be fertilized or stimulated by a depolarizing agent such as an excess of KCl, in contrast to the situation found in Crassostrea gigas. We have performed a comparative study between the two situations found in these species. In vitro, both of these oocytes can be triggered to reinitiate meiosis following a treatment by serotonin which promotes an intracellular calcium surge. Ruditapes and Crassostrea oocytes further arrest in metaphase I, at which stage they can be either activated by sperm or by excess KCl. These treatments trigger an intracellular calcium increase. This suggests that functional voltage-operated Ca2+ channels are expressed in Ruditapes during the course of maturation between prophase and metaphase I. Results obtained using pharmacological tools and direct binding of specific dihydropyridines, strongly suggest that these channels are dihydropyridine-sensitive calcium channels. In Ruditapes they become functional after 5-HT stimulation, their number increasing before GVBD. In Crassostrea the dihydropyridine-sensitive Ca2+ channels are already present at prophase stage and their density is constant from prophase to metaphase I. Moreover, we have shown for Ruditapes and Crassostrea that: 1) the addition of 10 microM of S(-)BayK8644, an agonist of dihydropyridine-sensitive calcium channels to metaphase-arrested oocytes releases them from metaphase block; and 2) incubating these oocytes with nicardipine, a potent blocker of dihydropyridine-sensitive Ca2+ channels, inhibits both their activation by excess KCl or fertilization. Taken together these data suggest that the absence of dihydropyridine-sensitive Ca2+ channels in the membrane of prophase-arrested oocytes of Ruditapes may account for their inability to be fertilized at this stage, while the presence of dihydropyridine sensitive Ca2+ channels in prophase-arrested oocytes of Crassostrea may explain their fertilizability at this stage. PMID- 11043417 TI - Production of stem-cell transplants according to good manufacturing practice. AB - Peripheral blood stem cells (PBSCs) are used for transplantation to reconstitute the hematopoietic system after high-dose chemotherapy. They are harvested from peripheral blood after mobilization by cytokines and/or chemotherapy. Further ex vivo manipulation steps (e.g., selection of CD34+ PBSCs, purging, expansion, and differentiation or gene transfer) can be performed. In 1997, more than 12,000 PBSC preparations were transplanted in Europe and the total number is steadily increasing [1]. To ensure quality and safety of the final cell products intended for clinical use, national and international guidelines and regulations have been issued. The implementation of a quality assurance (QA) program including the principles of good manufacturing practice (GMP) and a quality control system is a major requirement. GMP regulations apply to all phases of cell collection, processing, and storage, and to documentation, training of personnel, and equipment of the cell processing laboratory. They have to be followed by pharmaceutical companies and medical doctors who are involved in PBSC processing at academic institutions. The complicated regulatory network for the manufacturing of cell products will help to standardize these procedures and ensure consistent quality and safety in the long term. This will be in the interest of patients and reduce risks of application of individual cell preparations. PMID- 11043418 TI - E-selectin and very late activation antigen-4 mediate adhesion of hematopoietic progenitor cells to bone marrow endothelium. AB - Adhesion of CD34+ hematopoietic progenitor cells (HPCs) to sinusoidal endothelium probably plays a key role in homing of transplanted CD34+ HPCs to the bone marrow (BM). We have investigated the role of various adhesion molecules in the interaction of purified CD34+ HPCs derived from BM or peripheral blood (PB) and a human BM-derived endothelial cell line. Adhesion of CD34+ HPCs to endothelial cells was measured with the use of a double-color flow microfluorimetric adhesion assay. In this assay, adhesion is measured under stirring conditions, simulating blood flow in sinusoidal marrow vessels. Adhesion of PB CD34+ cells to human BM endothelial cells (HBMECs) was observed only after interleukin (IL)-1beta prestimulation of the endothelial cells. This adhesion was strongly increased after addition of phorbol-myristate acetate (PMA). Adhesion of PB CD34+ cells to IL-1beta-prestimulated HBMECs was inhibited by blocking monoclonal antibodies (mAbs) against E-selectin and by neuraminidase treatment of the PB CD34+ cells. mAbs against very late activation antigen (VLA)-4 inhibited adhesion only when the E-selectin-mediated interaction was prevented. No clear inhibiting effect was found with blocking mAbs against beta2-integrins. Stimulation with the beta1 integrin-activating mAb, 8A2, induced adhesion of CD34+ cells to endothelial cells. In conclusion, stimulation of both endothelial cells and CD34+ HPCs is necessary for adhesion of CD34+ HPCs to endothelial cells. We furthermore demonstrated that E-selectin and VLA-4 mediated this adhesion. PMID- 11043419 TI - Induction of apoptosis using 2',2' difluorodeoxycytidine (gemcitabine) in combination with antimetabolites or anthracyclines on malignant lymphatic and myeloid cells. Antagonism or synergism depends on incubation schedule and origin of neoplastic cells. AB - Induction of apoptosis in vitro using gemcitabine (dFdC) in combination with cladribine (2-CdA) and other cytotoxic drugs on malignant mononuclear cells (MNCs) of patients with acute myeloid leukemia (AML, n=20) and chronic lymphocytic leukemia (CLL, n =20) in myeloid (HL60, HEL) and lymphatic cell lines (HUT78, JURKAT) was investigated using different incubation conditions (simultaneous and consecutive). Furthermore, the influence of dFdC on the level of intracellular metabolites of 2-CdA was studied using high-performance liquid chromatography (HPLC). Apoptosis was evaluated using flow cytometry with 7 aminoactinomycin D. In MNCs of patients with CLL, dFdC + 2-CdA showed an antagonistic effect when applied simultaneously. This antagonism was reduced by consecutive application. The combination of dFdC with doxorubicin was synergistic, independent of incubation schedule. In blasts from newly diagnosed patients with de novo AML, all drug combinations (dFdC+2-CdA, doxorubicin, or cytosine arabinoside) were antagonistic by simultaneous incubation. Reduced antagonism or even synergism was shown (P<0.001) by consecutive incubation. The simultaneous combination of dFdC with 2-CdA in all tested cell lines resulted in a competitive inhibition on the rate of apoptosis. By changing the incubation period to a consecutive schedule, the antagonism was diminished or synergism of apoptosis was measured (P< 0.001). Using similar incubation conditions, these experiments were supported by HPLC measurement of intracellular metabolites of 2 CdA influenced by dFdC application. In conclusion, we demonstrated that the efficacy of dFdC in vitro in combination with other cytotoxic drugs depends on the incubation condition and on the origin of neoplastic cells (lymphatic vs myeloid). The data suggest that simultaneous combination therapy with purine and pyrimidine analogues may not improve the clinical efficacy of one or the other drug administered alone. PMID- 11043420 TI - IDEC-C2B8 (Rituximab) anti-CD20 antibody treatment in relapsed advanced-stage follicular lymphomas: results of a phase-II study of the German Low-Grade Lymphoma Study Group. AB - PURPOSE: The current study was initiated to assess the clinical efficacy and side effects of rituximab in patients with relapsed advanced stage follicular lymphoma. PATIENTS AND METHODS: The study was performed as an open-label non randomized multicenter phase-II trial and included patients older than 18 years of age with relapsed advanced-stage follicular lymphomas (FL) grades I and II, according to the REAL classification, or with centroblastic/centrocytic (CB/CC lymphomas according to the Kiel classification. Four weekly doses of 375 mg/m2 rituximab were applied. RESULTS: 38 patients from eight centers were included between January 1997 and January 1998 and were evaluable for response and toxicity on an intention to treat basis. The median age was 55 years (range 26-75 years). Thirteen patients (35%) were in first relapse, 11 patients (30%) in second, and 13 patients (35%) in third relapse. The median time between primary diagnosis and study entry was 4.6 years (range 0.9-14.7 years). Twenty-three patients tolerated the application of rituximab without adverse events; in 13 cases the infusion rate had to be reduced because of side effects; in two patients the application was stopped because of pharyngeal edema and anaphylactoid reaction. The most frequent side effects were fever (13 patients) and rigor (13 patients); 65% of the side effects were observed after the first infusion. Twenty grade-III/IV side effects were considered to be related to treatment: lymphocytopenia (3), granalocytopenia (1), thrombocytopenia (2), fever (1), hyperglycermia (1), venous thrombosis (1), syncope (1), plasmatic coagulation disorder (1), shortness of breath (2), photosensitivity (1), cardiac failure (1), chills (1), sepsis (1), tumor lysis (1), anemia (1), and pharyngeal edema (1). Eight patients were not eligible for assessment of response because of non-follicular subtypes of low-grade lymphomas (n =6) or early termination of therapy at the first infusion because of severe side effects (n =2). From the 30 evaluable cases with follicular lymphomas, five patients achieved a complete remission (CR) (17%), nine patients a partial remission (PR) (30%), and two patients a minor response (MR) (7%). The overall response rate was 47%. The median time to treatment progression (TTP) was 201 days (range 64-293 days), with five patients experiencing long-lasting remissions of 214-293 days duration. In three patients, the rituximab-induced remission exceeded the preceding progression-free interval substantially. Bulky disease (P=0.058) and/or bone-mar row involvement (P=0.046) were associated with poor response. CONCLUSION: This study confirms the moderate treatment-related toxicity and the high antilymphoma activity of rituximab in patients with relapsed follicular lymphoma. Further studies are needed to determine the role of rituximab in the first-line treatment of these disorders and its combination with conventional chemotherapy. PMID- 11043421 TI - Impact of pre-induction therapy leukapheresis on treatment outcome in adult acute myelogenous leukemia presenting with hyperleukocytosis. AB - Acute myeloid leukemia (AML) presenting with hyperleukocytosis is generally of poor prognosis due to an increased early death rate and a lower response to initial chemotherapy. Between April 1985 and December 1995, all patients with newly diagnosed AML admitted to our institution with an initial white blood cell (WBC) count greater than 100 x 10(9)/l were scheduled to undergo leukapheresis. This represented 53 patients (median age 59 years, range 16-78 years) who underwent from 1 to 4 sets of leukapheresis (median 1). The median initial WBC count was 160 x 10(9)/l (range 100-480 x 10(9)/l). Morphologic subtypes, according to the French-American-British classification, showed 3 M0, 16 M1, 6 M2, 10 M4, 16 M5, and 2 unclassified cases of AML. In 21 patients (40%), leukapheresis did not reduce their WBC counts significantly, while 32 patients (60%) achieved a WBC count of less than 100 x 10(9)/l (median 71 x 10(9)/l) after leukapheresis. Analysis of cell cycle was performed on bone marrow (BM) and peripheral blood leukemic cells before and after leukapheresis in three cases. In two of those cases, a recruitment of BM leukemic cells in the S phase was observed after leukapheresis. The median WBC count at the time of starting chemotherapy was 85 x 10(9)/l (range 23-264 x 10(9)/l). Complete remission was achieved in 55% (95% confidence interval 40-68%). Early death occurred in two cases. Median disease-free survival was 10 months, while median overall survival was 8 months. In this study, early death rate is lower than data previously published in the literature and almost all patients could receive chemotherapy. This might suggest a benefit of initial leukapheresis in the treatment of AML presenting with hyperleukocytosis. PMID- 11043423 TI - c-myc overexpression is not mandatory in aggressive-phase multiple myeloma with Burkitt's type translocation. AB - This report concerns a case of aggressive-phase multiple myeloma (AGMM) with Burkitt's type translocation t(8;14)(q24;q32), detected by Giemsa-banding. Double color fluorescence in situ hybridization identified the breakpoint on 8q24 at a comparatively centromeric site, which was at least 300 kb and possibly 600 kb distant from the c-myc coding region. The breakpoint on 8q24 of the present case was far removed from that seen in other B-cell neoplasms with t(8;14)(q24;q32). Despite the presence of t(8;14)(q24;q32), neither rearrangement nor overexpression of the c-myc gene was observed in this case. Although our case may be a special case of multiple myeloma, it nevertheless suggests that overexpression of c-myc is not mandatory in an AGMM patient with Burkitt's type translocation. t(8;14)(q24;q32) which was seen in our case represents one of the first to be mapped at more than 300 kb 5' of c-myc. It should also be noted that this result could mean that a centromeric boundary 5' of c-myc exists where the influence of the immunoglobulin (Ig) H enhancer on c-myc transcription is not effective. PMID- 11043422 TI - A transforming growth factor-beta1-mediated bystander immune suppression could be associated with remission of chronic idiopathic thrombocytopenic purpura. AB - Bystander immune suppression has been demonstrated in experimental models of oral immune tolerance induction. This phenomenon is associated with expression of transforming growth factor (TGF)-beta1 and T-helper cell (Th) 2 cytokines. We have studied serum levels of Th cytokines and B- and T-lymphocyte subsets in chronic idiopathic thrombocytopenic purpura (ITP), a disorder in which the production of platelet autoantibodies might be caused by a cytokine network dysregulation. Forty-six patients with ITP were separated into three groups depending on the platelet count (pltc): (1) < 50 x 10(9)/l, (2) 50-150 x 10(9)/l and (3) > 150 x 10(9)/l. We found significantly elevated plasma levels of the Th3 cytokine TGF-beta1 in patients with pltc >150x10(9)/l (23.5+/-2.8ng/ml), compared with patients with pltc <50x10(9)/l (2.3+/-0.6 ng/ml; P<0.0001), patients with pltc 50-150x 10(9)/l (7.2+/-1.7 ng/ml; P<0.0001) and healthy volunteers (9.8+/ 1.3 ng/ml; P<0.01). The serum levels of the Thl cytokines interleukin (IL)-2 and interferon (IFN)-y were below the detection limits of the assays. Likewise, the Th2 cytokine IL-4 was not detectable or was very low both in patients and controls. The serum levels of IL-10, a Th2 cytokine, were within the assay range and patients with pltc <50 x 10(9)/l had significantly lower levels (0.6+/-0.1 pg/ml) than both patients with pltc 50-150 x 10(9)/l (1.8 +/- 0.1 pg/ml; P<0.005) and healthy volunteers (1.4+/-0.1 pg/ml; P<0.005). Furthermore, patients with pltc <50 x 10(9)/l and splenectomised patients had significantly higher levels of CD4 + CD25 + activated T cells [26.2 +/- 14.8% (P<0.05) and 26.7+/-11.9% (P<0.005), respectively] than healthy controls (16.5+/-4.0%). Also, the number of natural killer (NK) cells among patients with pltc >150 x 10(9)/l were significantly elevated (26.6+/-16.0%; P<0.05) compared with controls (17.4+/ 7.6%). In conclusion, our data corroborate previous findings of elevated numbers of activated T cells in chronic ITP patients with active disease, but neither a clear-cut Th1 nor a Th2 serum cytokine profile could be established. However, ITP in remission was associated with elevated TGF-beta1, which might be a part of a bystander immune suppression. We propose that the effect of possible expression of TGF-beta1 by oral immune tolerance induction deserves to be explored in ITP patients with an active disease. PMID- 11043424 TI - Aggressive natural killer cell leukemia/lymphoma: a comprehensive cytogenetic study by spectral karyotyping. AB - A 38-year-old male presented with fever and hepatosplenomegaly. Cells that had infiltrated to the bone marrow were consistent immunophenotypically and genotypically with natural killer (NK) cells. Oligoclonal Epstein-Barr virus infection was detected in the bone marrow cells. The patient was diagnosed as a case of aggressive NK cell leukemia/lymphoma. Combined chemotherapy was not effective and death occurred shortly after presentation. Although the karyotype of this case was too complicated to be accurately identified only by G-banding, spectral karyotyping (SKY) analysis not only identified all chromosomal materials of unknown origin, but also detected the cryptic translocation on the apparently normal chromosome. Moreover, SKY analysis identified der(4)t(4;14)(q12;q11.2). The chromosomal band 14q11.2 is a recurring breakpoint in T-cell non-Hodgkin's lymphoma, and is also the locus of the delta chain of the T-cell receptor. To our knowledge, t(4;14)(q12;q11.2) in T-cell or NK-cell malignancies has not been previously reported. PMID- 11043425 TI - Fatal cardiac arrhythmia after infusion of dimethyl sulfoxide-cryopreserved hematopoietic stem cells in a patient with severe primary cardiac amyloidosis and end-stage renal failure. AB - Amyloidosis (AL) is a rapidly fatal plasma cell dyscrasia causing progressive multiorgan failure. Recently, substantial improvement of survival was reported following high-dose chemotherapy with peripheral blood stem cell (PBSC) rescue. We describe a patient with AL with severe cardiac and renal involvement who received high-dose melphalan followed by fractioned autologous PBSC transplantation (455 ml on day 1 and 350 ml on day 2). Immediately after the second infusion of the PBSCs, life-threatening cardiac arrhythmias occurred and, despite intensive treatment, the patient died less than 24 h later. The infusion of cryopreserved PBSCs may be associated with complications, including cardiac toxicity. Dimethyl sulfoxide (DMSO) is the most frequently used cryopreservation agent. In the present case, we suggest that DMSO could have played an important role in causing the fatal cardiac arrhythmias. The mechanisms of the cardiovascular effects of DMSO and the possible preventive measures are discussed. Given the poor prognosis of AL and the promising results of dose intensive chemotherapy with autologous PBSC transplantation, careful patient selection and intensive monitoring are mandatory in order to further pursue this therapeutic approach. PMID- 11043426 TI - Fatal thrombotic thrombocytopenic purpura as a rare complication following allogeneic stem cell transplantation. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare disease which, together with hemolytic uremic syndrome, is subsumed under thrombotic microangiopathy. After stem cell transplantation (SCT), this syndrome represents a possibly fatal complication with a higher incidence in allogeneic SCT than in autologous SCT. Although plasmapheresis offers an encouraging treatment modality in classic TTP, this seems less effective in bone marrow transplant-associated microangiopathy. This is probably due to a different etiology. We present a case of transplant associated TTP with a fatal outcome despite multiple courses of plasmapheresis. PMID- 11043427 TI - Primary hepatic or splenic diffuse large B-cell lymphoma and hepatitis C virus infection: a non-fortuitous association? PMID- 11043428 TI - Prader-Willi psychiatric syndrome and Velo-Cardio-Facial psychiatric syndrome. AB - Prader-Willi psychiatric syndrome and Velo-Cardio-Facial psychiatric syndrome: Similar to the studies on behavioural phenotypes, it is suggested to more rigorously promote the investigation of psychopathological phenotypes. The psychopathological profile in patients with Prader-Willi Syndrome (PWS) or Velo Cardio-Facial Syndrome (VCFS) appears to be not classifiable within the current nosological systems. On a descriptive level, PWS-psychotic states show similarities with the cycloid psychoses, but VCFS psychosis does not. It is therefore advocated to adopt the notion of a brain-structure phenotype as well as that of a syndrome-specific psychiatric disorder. PMID- 11043429 TI - Prenatal echographic diagnosis of laryngeal atresia as part of a multiple congenital anomalies (MCA) syndrome. AB - Prenatal echographic diagnosis of laryngeal atresia as part of a multiple congenital anomalies (MCA) syndrome: In this report we present the prenatal second trimester echographic diagnosis of laryngeal atresia in a male fetus with multiple associated congenital anomalies: oesophageal atresia, crossed fused ectopy of the right kidney, mild cutaneous syndactyly of fingers III-V and toes II-III, distinct facial appearance and single umbilical artery. Bilateral voluminous echogenic lungs were the major echographic diagnostic sign. The associated multiple congenital anomalies were not diagnostic for a distinct, recognizable multiple malformation syndrome. PMID- 11043430 TI - De novo translocation (2;18)(q21;q22) in a child with severe epilepsy, developmental delay and mild dysmorphism. AB - De novo translocation (2;18)(q21;q22) in a patient with severe epilepsy developmental delay and mild dysmorphism: We report on a patient presenting with severe epilepsy, hypotonia, developmental delay, blepharophimosis, low-set ears, camptodactyly and tapering fingers, and cutaneous syndactyly of toes II and III of the right foot. The MRI showed some loss of volume of the white matter and delayed myelination, no other specific anomalies were present. Chromosome analysis revealed a translocation involving chromosomes 2 and 18, which was characterized further by FISH using band-specific probes. The possibility of a submicroscopic deletion is discussed and the patient is compared with patients reported in the literature with either 2q21 or 18q22 deletion. PMID- 11043431 TI - Reproductive follow-up of carriers of familial reciprocal balanced translocations involving chromosome 9 and comparison with predicted outcome. AB - Reproductive follow-up of carriers of familial reciprocal balanced translocations involving chromosome 9 and comparison with predicted outcome: Chromosome 9 is commonly implicated in reciprocal translocations (rcp). Twenty-seven families segregating rcp involving chromosome 9 were selected with the aim of comparing the theoretical risk of Mental Retardation with Congenital Anomalies (MCA/MR) calculated according to Human Cytogenetics Forum with the observed reproductive follow-up. The 27 families include 157 subjects. The reproductive follow-up showed that the majority of mothers underwent full-term pregnancies (88/130), and that there were 37 spontaneous and five voluntary abortions. Eighty-one subjects were karyotyped: 18 had a normal karyotype, 50 carried an rcp, ten had an unbalanced rcp-related karyotype and three an abnormal rcp-unrelated karyotype. Of the 88 live-born individuals, seven had an abnormal rcp-related karyotype with partial chromosome 9 trisomy (four cases) or partial 9p monosomy (three cases), and 48 were rcp carriers, two of whom also presented additional anomalies. The evaluation of reproductive outcomes in the 27 families studied revealed good concordance between the Human Cytogenetics Forum predictions and the observed follow-up in relation to the most probable mode of unbalance at birth, and the higher risk of MCA/MR in rcp carriers with unbalanced live-borns in comparison with those generating healthy progeny PMID- 11043432 TI - Precarious acrocentric short arm in prenatal diagnosis: no chromosome 14 polymorphism, but trisomy 17p. AB - Precarious acrocentric short arm in prenatal diagnosis: no chromosome 14 polymorphism, but trisomy 17p: We report on a girl with multiple congenital abnormalities and a prenatally diagnosed 46,XX,14p+ de novo karyotype. Fluorescence in situ hybridization (FISH) demonstrated that the extra material on the short arm of chromosome 14 was not just a polymorphism, but that it originated from chromosome 17. The phenotypic findings of this patient with pure trisomy 17p are compared with those of ten previously published cases. PMID- 11043433 TI - Two sibs with duplication of 4q31-->qter due to 3:1 meiotic disjunction and mild phenotype. AB - Two sibs with duplication of 4q31-->qter due to 3:1 meiotic disjunction and mild phenotype: Clinical and cytogenetic findings in two sibs with partial duplication of 4q31.3-->qter and 21q11.2-->pter are reported. These patients are rare cases of reoccurrence of those partial trisomies due to 3:1 segregation of a maternal balanced translocation. A review of the literature reporting cases of trisomy of the 4q31-->qter segment is also made; previously reported cases mostly in addition have deletions of other chromosomes resulting from adjacent segregation of balanced translocation. The findings of our study confirm the high risk for offspring with unbalanced rearrangements in women with reciprocal translocation involving acrocentric and short chromosome segments. The study also points out that duplication of 4q31-->qter may go along with only mild phenotypic findings if there is no significant additional aneuploidy of the other chromosome involved in the rearrangement. PMID- 11043434 TI - A MELAS phenotype and a paternal inherited inversion of chromosome 10 in a female patient. AB - A MELAS phenotype and a paternal inherited inversion of chromosome 10 in a female patient: We describe a patient suffering from encephalomyopathy with overlapping symptoms, including MELAS and Kearn-Sayre syndrome features. Mutations in tRNA LEU (UUR) were not found in mtDNA of blood cells, suggesting a different genetic defect. Cytogenetic studies revealed a paternal inherited pericentric inversion of chromosome 10 (p13;q22) pat. Although the presence of the same inversion in the father and in the apparently asymptomatic sister does rather suggest that the concurrence of the mitochondrial disease in the patient was due to chance, some alternative explanations to associate both events might be proposed. PMID- 11043435 TI - A case with 47,XXY,del(11)(q23) karotype-coexistence of Jacobsen and Klinefelter syndromes. AB - A case with 47,XXY, del(11)(q23) karyotype-coexistence of Jacobsen and Klinefelter syndromes: A two-year-old dysmorphic male child was found to have 47,XXY,del(11)(q23) karyotype. Domination of the clinical features of Jacobsen syndrome was observed: mild mental retardation, trigonocephaly, ptosis, downward slanting palpebral fissures, low set ears, carp-shape mouth and micrognathia. Transient thrombocytopenia and leukopenia were also present. Over the following five years gynecomastia and eunuchoid body proportions became evident as clinical features of Klinefelter syndrome. This is the first description of the coexistence of both syndromes. PMID- 11043436 TI - Conductive hearing loss and multiple pre- and supra-auricular skin defects: a variant example of the Branchio-Oculo-Facial syndrome. AB - Conductive hearing loss and multiple pre- and supra-auricular skin defects: a variant example of the Branchio-Oculo-Facial Syndrome: We describe a 3-year-old girl with bilateral severe conductive hearing loss and multiple bilateral supra- and preauricular defects with unusual overlying thin skin. The severe conductive hearing loss is due to abnormal configuration of the ossicular chain. Multiple pre- and supra-auricular skin defects have been described in the Branchio-Oculo Facial syndrome, and we propose that the findings in our present patient may be a variant manifestation of this syndrome. PMID- 11043437 TI - Hypersplenism and portal hypertension with vena porta thrombosis in cartilage hair hypoplasia (metaphyseal chondrodysplasia, McKusick type, MIM *250250) PMID- 11043438 TI - Penile agenesis as an isolated malformation: a rare example of sexual ambiguity at birth. PMID- 11043439 TI - A stereological study of long-term regeneration of rat severed sciatic nerve repaired by means of muscle-vein-combined grafts. AB - This study is a stereological analysis, by the 2-D dissector method, on the long term regeneration of myelinated nerve fibers of the rat sciatic nerve repaired by muscle-vein-combined graft, a surgical technique that has been shown to be a valid tool for the repair of peripheral nerve defects with substance loss. Quantitative analysis showed that the total number and mean density of regenerated myelinated nerve fibers was significantly higher than in control nerves. The contrary was true for fiber mean size. The morpho-quantitative parameters of regenerated fibers from nerves repaired by the muscle-vein-combined graft were similar to those observed in rats where nerve defects were repaired by direct nerve suture thus confirming the validity of this surgical technique. PMID- 11043440 TI - On the bulky appliances and artero-venous anastomoses in the vascular district of the base of the brain. AB - It has been studied the vascular territory of the base of the brain and it has been pointed out the presence of characteristical structural arrangements of the vasal wall and artero-venous anastomoses. The just mentioned characteristical structures are placed in points where a vessel divides itself or creates a collateral branch and their functional engagement is target-oriented to control the blood flux. PMID- 11043441 TI - The vertebral canal and the lateral recess of the lombar tract: an anatomo radiologic study. AB - To address this work, we studied the vertebral canal and the lateral recess at the L1 to L5 level. The importance of this tract of vertebral column lies in the frequent pathologies involving this anatomical region. In fact, the diagnosis of any pathological condition requires the knowledge of the normal status of that anatomical district. In particular, in this study, we suggested the presence of a relation between the diameter of vertebral canal and lateral recess. In addition, the age of the subjects did not seem related to these diameters. In conclusion, we confirmed also that the living-obtained data have a practical utility for clinical and surgical applications. PMID- 11043442 TI - Age-related changes in skeletal muscle fiber composition in two swine muscles. AB - This study focuses the aging-related modification of skeletal fiber types in two skeletal muscles of different-age swine (6 and 18 month). Rectus abdominis and vastus medialis were employed. It was performed an immunohistochemical staining for slow fibers and it was made a quantitative evaluation, using an automatic interactive image analysis system. The percentage of slow fibers decreased in adult swine. Moreover, slow fibers in rectus abdominis were less numerous than in vastus medialis. Aging and muscle function are two important factors able to modify fiber types. Morphometric analyses can ascertain this modification for diagnostic or nourishmental purposes. PMID- 11043443 TI - Immunocytochemical expression of protein kinase C isoenzymes alpha, delta, epsilon and zeta in differentiating chick chondrocytes in vitro. AB - In our previous work we have investigated the expression of the serine-threonine kinase protein kinase C (PKC) in the vertebral column of mouse foetuses. In the present work we would verify the expression of four PKC-isoenzymes (alpha, delta, epsilon, zeta) in two distinct phases of the chondrogenesis and the endochondral osteogenesis in vitro. We performed primary cultures of chondrocytes collected from tibiae of 6-day old chick embryos. This cells were cultured for 20 days and than collected on coverslips (stage 1 culture). Other cells of the stage 1 were undergone further differentiation towards the phenotype of osteoblast-like cells (stage 2 culture), in accord to the protocol of Descalzi Cancedda et al. (1992). In stage 1 culture, PKC-epsilon was the most expressed isoform, whereas PKC-alpha exhibited the least intense positivity. In stage 2 culture, PKC-alpha was the most expressed isoform, whereas a marked decrease of PKC-epsilon expression was detected compared to stage 1. No relevant differences were evidenced as regards,the expression of PKC-zeta between the two considered cell culture stages. On these bases, it could be reasonable that these PKC-isoenzymes may be involved at different levels in chondrocytes differentiation as well as in the endochondral ossification process. PMID- 11043444 TI - Lectin histochemistry for detection of the oligosaccharides in the testis of the chick embryo. AB - The oligosaccharidic content, distribution and changes of the glycoconjugates in the testis of the chick embryo, from the 8th day of incubation to hatching, were studied using a battery of seven HRP-conjugated lectins (DBA, SBA, PNA, ConA, WGA, LTA, and UEA I). Our findings showed that ConA and WGA appeared to characterize the spermatogonia during their differentiation, maturation, migration and meiosis. In the Sertoli cells, a change of localization of staining with ConA and WGA was revealed during the differentiation and maturation of these cells. The basal membrane was characterized by the reactivity with ConA and WGA from the early stages of incubation. ConA, WGA and PNA reacted with the endothelial cells of the testis for the whole period of incubation considered. Moreover, the interstitial cells, since their appearance, showed reactivity with ConA, WGA and PNA at the plasma membrane and the cytoplasm. PMID- 11043445 TI - Corrosion cast of the vascularization of Mugil Cephalus gills. AB - The spatial organization of the microcirculation in gills of Mugil Cephalus, was examined by scanning electron microscopic analysis of corrosion cast prepared by intravascular injection of methyl methacrylate. The afferent branchial artery originates from the ventral aorta and gives rise to afferent filamental artery. From the medio-lateral wall of the afferent filamental artery, afferent lamellar arterioles originate which supply one or more lamellae. The lamellar efferent arterioles, which drain the blood coming from the lamellae into the efferent filamental arteries, continue with the efferent branchial arteries and then the dorsal aorta. The techniques used so permitted to evaluate the structure and the interrelationships of the vascular pathways, explaining the regulation and the distribution of the blood flow in the gills. PMID- 11043446 TI - Positive inotropic effects of imidazoline derivatives are not mediated via imidazoline binding sites but alpha1-adrenergic receptors. AB - Imidazoline-binding sites are non-adrenergic receptors and classified into I11/I2 subtypes. There is strong evidence that I1-binding sites, located in the rostro ventrolateral medulla, are involved in regulation of blood pressure. However, less is known about the peripheral participation of I1-binding sites in cardiovascular reactions. Therefore, the aim of this study was to investigate whether specific imidazoline derivatives influence myocardial contractility and whether imidazoline binding sites are expressed in rat heart. Agmatine, clonidine and idazoxan failed to alter inotropy in left atria within the whole concentration range tested (1 nM - 100 microM), whereas cirazoline (1- 100 microM) and moxonidine (100 microM) increase inotropy by about 20-30%. After preincubation with the alpha1-adrenoceptor antagonist prazosin, the cirazoline and moxonidine stimulated inotropy was antagonized, indicating more an alpha1 adrenergic and less an imidazoline binding site mediated mechanism. Radioligand binding studies in membranes of left ventricles using [3H]-clonidine to specify I1-binding yielded KD values of 12.7 microM, confirming the functional results of an absence of I1-binding sites in ventricles of rats. However, the existence of low affinity I2-binding sites determined by [3H]-idazoxan labeling could not be excluded since a KD of 0.5 microM was calculated and since competition studies with guanabenz (Ki = 0.1 microM), clonidine (Ki = 58.1 microM) and moxonidine (Ki = 129 microM) confirmed the specificity of the I2-binding. PMID- 11043447 TI - Pharmacological characterization of a novel sulfonylureid-pyrazole derivative, SM 19712, a potent nonpeptidic inhibitor of endothelin converting enzyme. AB - We describe the pharmacological characteristics of SM-19712 (4-chloro-N-[[(4 cyano-3-methyl-1-phenyl-1H-pyrazol-5-yl)amino]carbonyl] benzenesulfonamide, monosodium salt). SM-19712 inhibited endothelin converting enzyme (ECE) solubilized from rat lung microsomes with an IC50 value of 42 nM and, at 10 - 100 microM, had no effect on other metalloproteases such as neutral endopeptidase 24.11 and angiotensin converting enzyme, showing a high specificity for ECE. In cultured porcine aortic endothelial cells, SM-19712 at 1 - 100 microM concentration-dependently inhibited the endogenous conversion of big endothelin-1 (ET-1) to ET-1 with an IC50 value of 31 microM. In anesthetized rats, either intravenous (1-30 mg/kg) or oral (10-30 mg/kg) administration of SM-19712 dose dependently suppressed the pressor responses induced by big ET-1. In acute myocardial infarction of rabbits subjected to coronary occlusion and reperfusion, SM-19712 reduced the infarct size, the increase in serum concentration of ET-1 and the serum activity of creatinine phosphokinase. The present study demonstrates that SM-19712 is a structurally novel, nonpeptide, potent and selective inhibitor of ECE, and SM-19712 is a valuable new tool for elucidating the pathophysiological role of ECE. PMID- 11043449 TI - Characterization of the ca2 + response mediated by activation of beta adrenoceptors in rat submandibular ducts. AB - The Ca2+ signaling mediated by activation of beta-adrenoceptors was studied in a purified preparation of ducts from rat submandibular glands. At concentrations above 1 nM, isoproterenol (ISO) caused a small but significant increase in cytosolic Ca2+ concentration ([Ca2+]i). The ISO-induced increase in [Ca2+]i was completely inhibited by the beta-adrenoceptor antagonist propranolol but not by the alpha-adrenoceptor antagonist phentolamine. Forskolin was able to mimic the Ca2+ response to ISO. These results suggest that the ISO-induced increase in [Ca2+]i in rat submandibular ducts is mediated by an accumulation of cAMP resulting from activation of beta-adrenoceptors. In the absence of extracellular Ca2+, ISO or forskolin caused a transient increase in [Ca2+]i, indicating Ca2+ mobilization from intracellular Ca2+ stores. Further, stimulation with ISO failed to mobilize Ca2+ after the depletion of intracellular Ca2+ stores by phenylephrine or carbachol, suggesting that the cAMP-mediated increase in [Ca2+]i is due to a Ca2+ release from inositol trisphosphate (IP3)-sensitive Ca2+ stores. As ISO did not stimulate a detectable production of IP3, the cAMP-mediated Ca2+ mobilization may be evoked by a mechanism different from activation of phosphoinositide hydrolysis. PMID- 11043448 TI - Protective effect of SM-19712, a novel and potent endothelin converting enzyme inhibitor, on ischemic acute renal failure in rats. AB - Effects of SM-19712 (4-chloro-N-[[(4-cyano-3-methyl- 1-1-phenyl- 1H-pyrazol-5 yl)amino]carbonyl] benzenesulfonamide, monosodium salt), a novel endothelin converting enzyme (ECE) inhibitor, on ischemic acute renal failure (ARF) in rats were examined in comparison with those of phosphoramidon, a conventional ECE inhibitor. ARF was induced by occlusion of the left renal artery and vein for 45 min followed by reperfusion, 2 weeks after contralateral nephrectomy. Renal function in ARF rats markedly decreased at 24 h after reperfusion. Intravenous bolus injection of SM-19712 (3, 10, 30 mg/kg) prior to the occlusion attenuated dose-dependently the ischemia/reperfusion-induced renal dysfunction. Histopathological examination of the kidney of ARF rats revealed severe renal damages such as tubular necrosis, proteinaceous casts in tubuli and medullary congestion, all of which were dose-dependently attenuated by SM-19712. Protective effects of phosphoramidon (10 mg/kg) on ARF-induced functional and tissue damages were less potent than that of the same dose of SM-19712. Endothelin-1 (ET-1) content in the kidney after the ischemia/reperfusion was significantly increased, being the maximum level at 6 h after reperfusion, and this elevation was completely suppressed by the higher dose of SM-19712. Our findings support the view that renal ET-1 plays an important role in the development of ischemia/reperfusion-induced renal injury. SM-19712 may be useful in the treatment of ischemic ARF. PMID- 11043450 TI - Effects of perinatal nicotine exposure on development of [3H]hemicholinium-3 binding sites in rat neonate brain. AB - In this study, [3H]hemicholinium-3 ([3H]HC-3) binding, which labels the presynaptic high affinity-choline transport sites, was examined in two brain regions, cerebral cortex and midbrain, of nicotine-treated and -untreated rat neonates. In nicotine-untreated neonates, [3H]HC-3 binding sites of cerebral cortex increased from 64 fmol/mg protein at postnatal day 7 to 142 fmol/mg protein at postnatal day 35. In nicotine-treated neonates, the development of [3H]HC-3 binding sites in cerebral cortex was significantly retarded, compared with control neonates on the 7th, 14th and 21st postnatal days. In parallel with this, the development of muscarinic receptor in cerebral cortex, which was detected by [3H]quinuclidinyl benzylate ([3H]QNB) binding, was also retarded by nicotine treatment. However, in midbrain, neither [3H]HC-3 nor [3H]QNB binding sites at postnatal day 14 was affected by nicotine treatment. These results strongly suggest that perinatal treatment with nicotine inhibits presynaptic and postsynaptic development of the cholinergic pathway in cerebral cortex but not in midbrain of rat neonate. PMID- 11043451 TI - In vitro and in vivo long term release of apomorphine from polymer matrices. AB - Since apomorphine actually reveals high efficacy in treatment of Parkinson's disease but only has a very short half life, it is of only limited clinical significance. To overcome this substantial disadvantage, drug application by long term delivery systems could be one possibility. Based on this background, ethylene vinyl acetate polymeric delivery systems were manufactured that differed in size, with either coated or uncoated surfaces, but were similar in apomorphine loading. Release from uncoated polymeric delivery systems followed first order kinetics, whereas coated polymeric delivery systems showed within the first 40 days a period of first order kinetics release, in which the release rate is approximately half that of the uncoated polymeric delivery systems, followed by a zero order kinetics release for more than 130 days with a daily release rate of 3.1 +/- 0.2 mg. In vivo release was investigated by determining plasma apomorphine concentrations after implanting polymeric delivery systems into the abdominal cavities of rats. Animals with uncoated polymeric delivery systems exhibited symptoms of an apomorphin overdosage within 20 days after surgery. Using coated polymeric delivery systems, a steady state plasma concentration of 15 ng/ml was observed, which was maintained over a period of 130 days after an initial period of high plasma concentrations. Based on our results, it is concluded that polymeric delivery systems might be an appropriate method for applying apomorphine for the treatment of Parkinson's disease. PMID- 11043452 TI - Cholecystokinin acts as an essential factor in the exacerbation of pancreatic bile duct ligation-induced rat pancreatitis model under non-fasting condition. AB - We examined the influence of 2 gut hormones involved in the enhancement of pancreatic exocrine secretion, secretin and cholecystokinin (CCK), in the exacerbation of pancreatitis. We also examined the role of the vagal system, which was considered to be a transmission route for these hormones. Our model of pancreatitis in the rat was prepared by pancreatic bile duct ligation (PBDL), which simultaneously ligated the pancreatic duct and the common bile duct. Serum amylase activity and histopathological changes in the pancreas were used as indices of pancreatitis. We also measured the volume of pancreatic juice, as well as the amylase activity and protein level of the pancreatic juice, as indices of increased pancreatic exocrine secretion. Two gut hormones were given 6 times at 1 h intervals. Administration of secretin (1-3 microg/kg, s.c.) did not influence serum amylase activity in rats with PBDL-induced pancreatitis. However, food stimulation and administration of CCK-8 (1 microg/kg, s.c.) increased serum amylase activity and promoted vacuolation of the pancreatic acinar cells in rats with PBDL-induced pancreatitis. Administration of atropine (3 mg/kg, s.c.) or a CCK1-receptor antagonist, Z-203 (0.1 mg/kg, i.v.), inhibited food-stimulated or CCK-8-induced (1 microg/kg, s.c.) enhancement of pancreatic exocrine secretion and exacerbation after the development of PBDL-induced pancreatitis. These results suggest that not secretin, which regulates the volume of pancreatic juice, but CCK, which regulates the secretion of pancreatic enzymes via the vagal system, plays an essential role in food-stimulated exacerbation after the development of pancreatitis. PMID- 11043453 TI - Food deprivation depletes gastric mucus glycoprotein in streptozotocin-induced diabetic rats. AB - Fasting causes gastric mucosal damage in streptozotocin (STZ)-induced diabetic rats, but its pathogenic mechanism remains to be elucidated. The aim of the present study was to investigate the alteration of gastric mucosal mucin, one of the gastric defensive factors against the development of such damage. Diabetes was induced in rats by intravenous injection of STZ (65 mg/kg). The experiments were performed using 4-week STZ-diabetic rats with blood glucose levels above 350 mg/dl. The amount of gastric mucus glycoprotein was determined by gel filtration, and the distribution of neutral and acidic mucins in the stomach epithelium was examined by histochemical analysis. In normal rats, 24-h fasting neither affected the gastric mucin content nor caused any macroscopic gastric mucosal injury. In contrast, starvation significantly reduced the amount of total gastric mucus glycoprotein prior to the formation of mucosal lesions in the STZ-diabetic rats. Nine hours after food deprivation, the gastric damage developed in about 70% of the diabetic rats, the amount of mucus glycoprotein markedly decreased, and both the neutral and acidic mucins diminished in the epithelium. Taken together, in STZ-diabetic rats, fasting by itself depletes gastric mucus glycoprotein, and this depletion may be involved in the pathogenic mechanism of the formation of gastric mucosal lesions. PMID- 11043454 TI - Effects of benidipine hydrochloride on cerebrovascular lesions in salt-loaded stroke-prone spontaneously hypertensive rats: evaluation by magnetic resonance imaging. AB - We determined possible protective effects of benidipine hydrochloride (benidipine), a dihydropyridine calcium antagonist, on cerebrovascular lesions in salt-loaded stroke-prone spontaneously hypertensive rats (SHRSP). The animals were orally treated with benidipine at 1, 3 and 10 mg/kg daily for 7 weeks, and their neurological symptoms, body weight changes, systolic blood pressure and cerebrovascular lesions on magnetic resonance imaging (MRI) were determined at various time points of treatment. Moreover, the brains of the rats that showed cerebrovascular lesions on MRI in the course of treatment or completed 7-week treatment were examined histopathologically. Control rats presented such symptoms as sedation, ataxia and aggressiveness, while their MRI analysis revealed high signals over wide areas from the occipital to frontal cortex and from the corpus callosum to external capsule. These high signal areas corresponded in location to edematous or softening lesions revealed by the histopathological observation. Treatment with benidipine at 3 and 10 mg/kg ameliorated neurological symptoms, significantly suppressing cerebrovascular damages on MRI. Benidipine at 3 mg/kg significantly decreased blood pressure for the first four weeks but it did not thereafter. These findings demonstrate that benidipine can protect salt-loaded SHRSP from cerebrovascular injury as assessed by MRI. PMID- 11043456 TI - Serotonin induces apoptosis in PGT-beta pineal gland tumor cells. AB - Serotonin-induced neuronal cell death has been implicated as a possible cause of neurodegenerative and neuropsychiatric disorders. To investigate the involvement of serotonin-induced apoptosis as a potential mechanism in the pathophysiology of serotonin-related diseases, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay, DAPI (4',6-diamidino-2-phenylindole) staining, TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling) assay and flow cytometric analysis were performed using the immortalized pineal cell line PGT-beta. Through morphological and biochemical analyses, it was demonstrated that cell death induced by serotonin in PGT-beta cells shows classic apoptotic features. These data suggest that serotonin induces apoptosis in PGT-beta cells. PMID- 11043455 TI - Protection by polaprezinc, an anti-ulcer drug, against indomethacin-induced apoptosis in rat gastric mucosal cells. AB - Polaprezinc [N-(3-aminopropionyl)-L-histidinato zinc] (PZ), an anti-ulcer drug, is a chelate compound consisting of zinc and L-carnosine. PZ has been shown to prevent gastric mucosal injury. In the present study, we investigated the inhibitory effect of PZ on indomethacin (IND)-induced apoptosis in a rat gastric mucosal cell line, RGM1. Pretreatment with PZ suppressed caspase-3 activation and subsequent apoptosis in the cells exposed to 500 microM IND in a dose-dependent manner, and 50 microM PZ exhibited the maximum inhibitory effect. Among PZ subcomponents, zinc but not L-carnosine played a pivotal role in this antiapoptotic function. PZ did not affect mitochondrial cytochrome c release upstream of caspase-3 activation in the IND-induced apoptotic signal pathway. Treatment with 500 microM IND evidently produced reactive oxygen species (ROS) in RGM1 cells. However, PZ did not scavenge ROS in IND-treated cells. Moreover, N acetylL-cysteine, a potent antioxidant, inhibited ROS generation but did not suppress apoptosis in RGM1 cells exposed to IND. These observations demonstrate a novel pharmacological action of PZ; i.e., that PZ, and in particular its zinc subcomponent, inhibits apoptosis via inhibition of caspase-3 activation but not antioxidant activity. PMID- 11043457 TI - Cardiac angiotensin II receptors are downregulated by chronic angiotensin II infusion in rats. AB - We studied the effect of chronic (7 days) angiotensin II infusion in a subpressor (200 ng/kg per minute) dose on angiotensin II receptors in the left ventricle in rats. Infusion of angiotensin II caused an elevation in systolic blood pressure after 3 days, usually to values of about 150 mmHg, and the increase continued during the drug administration. The number of angiotensin II type 1 and angiotensin II type 2 receptors was significantly decreased by 20-30% in the angiotensin II-infused left ventricular membranes without affecting the affinity. Thus, these data suggest that angiotensin II may regulate the number of its own receptors in rat left ventricles. PMID- 11043458 TI - Repeated adenosine pre-treatment potentiates the acute effect of methamphetamine in rats. AB - Adenosine was intraperitoneally (i.p.) injected to Wistar rats every 3 days with a total of 5 administrations. After a 7-day withdrawal, the animals were challenged with methamphetamine (0.5 mg/kg, i.p.). The effect of methamphetamine on locomotor activity was significantly potentiated by repeated adenosine pretreatment. Moreover, methamphetamine-induced dopamine release was also increased in the striatum. Methamphetamine-induced hyperactivity and dopamine release were significantly potentiated by repeated pretreatment of an adenosine A1 agonist, N6-cyclohexyladenosine (0.5 mg/kg, i.p.). These results suggest that the acute effect of methamphetamine is potentiated by repeated pre-treatment of adenosine via adenosine A1 receptors. PMID- 11043459 TI - Captopril increases the affinity of bradykinin receptor binding sites in bovine coronary arterial endothelial cells. AB - In a radioligand binding study using bovine coronary artery endothelial cell membranes, captopril changed a single bradykinin (BK) binding site (Kd = 1.77 nM, Bmax = 60.2 fmol/mg protein) to high- (Kd = 0.68 pM, Bmax = 17.7 fmol/mg protein) and low- (Kd = 1.00 nM, Bmax = 72.5 fmol/mg protein) affinity binding sites. This effect was reversed by GppNHp. Captopril also enhanced BK-induced endothelium dependent relaxation in saponin-treated coronary rings, and GppNHp partially suppressed this enhancement. These results suggest that captopril may affect BK receptors that couple to G-proteins. PMID- 11043460 TI - Effect of intracerebroventricular administration of soybean lecithin transphosphatidylated phosphatidylserine on scopolamine-induced amnesic mice. AB - The effect of intracerebroventricularly administered soybean lecithin transphosphatidylated phosphatidylserine (SB-tPS) on memory impairment was evaluated by a passive avoidance task. SB-tPS significantly prolonged the step through latency induced by scopolamine treatment as in our previous report where SB-tPS was orally administered. The same doses of soybean phosphatidylcholine were ineffective. This result indicates that SB-tPS can act on the brain without any peripheral modification. PMID- 11043461 TI - Effect of mexiletine on thermal allodynia and hyperalgesia in diabetic mice. AB - The antinociceptive effect of mexiletine in diabetic mice was examined. Tail flick latencies at heat intensity of 35 and 50 V in diabetic mice were shorter than those in non-diabetic mice. In diabetic mice, mexiletine increased the tail flick latency at 35 V to the level observed in non-diabetic mice. The tail-flick latency at 50 V in diabetic mice, but not in non-diabetic mice, was increased by pretreatment with capsaicin (0.56 nmol, i.t., 24 h). The antinociceptive effect of mexiletine in diabetic mice was reduced by capsaicin. These results suggest that the mexiletine-induced antinociception in diabetic mice involves the inhibition of the nociceptive transmission of capsaicin-sensitive primary afferent fibers. PMID- 11043462 TI - The effects of soybean transphosphatidylated phosphatidylserine on cholinergic synaptic functions of mice. AB - The effects of soybean transphosphatidylated phosphatidylserine (SB-tPS) on cholinergic synaptic functions were investigated using cerebral cortical synaptosomes from mice. Treatment of the synaptosomes with SB-tPS increased high K+-induced acetylcholine (ACh) release in a bell-shaped, dose-dependent manner without affecting ACh synthesis. SB-tPS (10 and 50microM) also enhanced synaptosomal synthesis of sn-glycero-3-phosphocholine, but did not affect phosphorylcholine synthesis. In contrast, the choline synthesis was significantly reduced as SB-tPS concentration increased. The present result that SB-tPS modified the cholinergic pathway can partly explain its nootropic functions. PMID- 11043463 TI - SK4 encodes intermediate conductance Ca2+-activated K+ channels in mouse urinary bladder smooth muscle cells. AB - The single channel current of intermediate conductance Ca2 +-activated K+ channel (IK channel) was measured in mouse urinary bladder myocytes (MBM), and the molecular basis of the channel was suggested to be the SK4 subtype by RT-PCR. Among Ca2+-activated K+ channel subtypes (SK2, SK3, SK4 and BK), the mRNAs of SK4 and BK were predominantly expressed in MBM. IK channel currents recorded from MBM showed: 38.7 pS slope conductance under symmetrical 140 mM K+ conditions, Ca2+ dependent activation, and blockade by charybdotoxin and econazole. These results strongly suggest that SK4 encodes IK channels in MBM. PMID- 11043464 TI - Contribution of fungal loline alkaloids to protection from aphids in a grass endophyte mutualism. AB - Fungal endophytes provide grasses with enhanced protection from herbivory, drought, and pathogens. The loline alkaloids (saturated 1-aminopyrrolizidines with an oxygen bridge) are fungal metabolites often present in grasses with fungal endophytes of the genera Epichloe or Neotyphodium. We conducted a Mendelian genetic analysis to test for activity of lolines produced in plants against aphids feeding on those plants. Though most loline-producing endophytes are asexual, we found that a recently described sexual endophyte, Epichloe festucae, had heritable variation for loline alkaloid expression (Lol+) or nonexpression (Lol-). By analyzing segregation of these phenotypes and of linked DNA polymorphisms in crosses, we identified a single genetic locus controlling loline alkaloid expression in those E. festucae parents. We then tested segregating Lol+ and Lol- full-sibling fungal progeny for their ability to protect host plants from two aphid species, and observed that alkaloid expression cosegregated with activity against these insects. The in planta loline alkaloid levels correlated with levels of anti-aphid activity. These results suggested a key role of the loline alkaloids in protection of host plants from certain aphids, and represent, to our knowledge, the first Mendelian analysis demonstrating how a fungal factor contributes protection to plant-fungus mutualism. PMID- 11043465 TI - Activation of the cAMP pathway in Ustilago maydis reduces fungal proliferation and teliospore formation in plant tumors. AB - In the corn smut fungus Ustilago maydis, mating of two haploid sporidia is a prerequisite for subsequent colonization of the host. Cyclic AMP (cAMP) and pheromone signals have been implicated in this developmental program. The cAMP pathway is also needed for subsequent fungal development in planta, as null mutants in any component of the pathway fail to form tumors. Here we show that moderate activation of the pathway conferred either by mutation in the Galpha subunit or by mutation in the regulatory subunit of the protein kinase A influences tumor morphology. In the resulting tumors, the amount of fungal material is drastically reduced and fungal development is arrested at the stage of sporogenic hyphae. We conclude that tight regulation of the cAMP pathway is crucial for fungal development within the plant but does not interfere with the tumor induction process. PMID- 11043466 TI - Stagonospora avenae secretes multiple enzymes that hydrolyze oat leaf saponins. AB - The phytopathogenic fungus Stagonospora avenae is able to infect oat leaves despite the presence of avenacoside saponins in the leaf tissue. In response to pathogen attack, avenacosides are converted into 26-desglucoavenacosides (26 DGAs), which possess antifungal activity. These molecules are comprised of a steroidal backbone linked to a branched sugar chain consisting of one alpha-L rhamnose and two (avenacoside A) or three (avenacoside B) beta-D-glucose residues. Isolates of the fungus that are pathogenic to oats are capable of sequential hydrolysis of the sugar residues from the 26-DGAs. Degradation is initiated by removal of the L-rhamnose, which abolishes antifungal activity. The D-glucose residues are then hydrolyzed by beta-glucosidase activity. A comprehensive analysis of saponin-hydrolyzing activities was undertaken, and it was established that S. avenae isolate WAC1293 secretes three enzymes, one alpha rhamnosidase and two beta-glucosidases, that carry out this hydrolysis. The major beta-glucosidase was purified and the gene encoding the enzyme cloned. The protein is similar to saponin-hydrolyzing enzymes produced by three other phytopathogenic fungi, Gaeumannomyces graminis, Septoria lycopersici, and Botrytis cinerea, and is a family 3 beta-glucosidase. The gene encoding the beta glucosidase is expressed during infection of oat leaves but is not essential for pathogenicity. PMID- 11043468 TI - Molecular analysis of thermoregulation of phaseolotoxin-resistant ornithine carbamoyltransferase (argK) from Pseudomonas syringae pv. phaseolicola. AB - The phaseolotoxin-resistant ornithine carbamoyltransferase (ROCT) and phaseolotoxin are produced by Pseudomonas syringae pv. phaseolicola at 18 degrees C but not at 28 degrees C. At 28 degrees C, the pathogen produces a protein(s) that binds (in vitro) to a 485-bp fragment (thermoregulatory region, TRR) from a heterologous clone from the pathogen genomic library, which in multiple copies overrides thermoregulation of phaseolotoxin production in wild-type cells (K. B. Rowley, D. E. Clements, M. Mandel, T. Humphreys, and S. S. Patil, Mol. Microbiol. 8:625-635, 1993). We report here that DNase I protection analysis of the 485-bp fragment shows that a single site is protected from cleavage by the protein in the 28 degrees C extract and that this site contains two repeats of a core motif G/C AAAG separated by a 5-bp spacer. Partially purified binding protein forms specific complexes with a synthetic oligonucleotide containing four tandem repeats of this motif. A 492-bp upstream fragment from argK encoding ROCT also forms specific complexes with the protein in the 28 degrees C crude extract, and a 260-bp subfragment from the TRR containing the binding site cross competes with the argk fragment, indicating that the same protein binds to nucleotides in both fragments. DNase I protection analysis of the fragment from argK revealed four separate protected sequence elements, with element III containing half of the core motif sequence (CTTTG), and the other elements containing similar sequences. Gel shift assays were done with DNA fragments from which one or all of the sites were removed as competitor DNAs against the argK probe. The results of these experiments confirmed that the binding sites (in argK) are necessary for the protein to bind to the argK fragment in a specific manner. Taken together, the results of studies presented here suggest that in cells of P. syringae pv. phaseolicola grown at high temperature argK may be negatively regulated by the protein produced at this temperature. PMID- 11043467 TI - Differential expression of two soybean apyrases, one of which is an early nodulin. AB - Two cDNA clones were isolated from soybean (Glycine soja) by polymerase chain reaction with primers designed to conserved motifs found in apyrases (nucleotide phosphohydrolase). The two cDNAs are predicted to encode for two, distinct, apyrase proteins of approximately 50 kDa (i.e., GS50) and 52 kDa (i.e., GS52). Phylogenetic analysis indicated that GS52 is orthologous to a family of apyrases recently suggested to play a role in legume nodulation. GS50 is paralogous to this family and, therefore, likely plays a different physiological role. Consistent with this analysis, GS50 mRNA was detected in root, hypocotyls, flowers, and stems, while GS52 mRNA was found in root and flowers. Neither gene was expressed in leaves or cotyledons. Inoculation of roots with Bradyrhizobium japonicum, nitrogen-fixing symbiont of soybean, resulted in the rapid (<6 h) induction of GS52 mRNA expression. The level of GS50 mRNA expression was not affected by bacterial inoculation. Western blot (immunoblot) analysis of GS50 expression mirrored the results obtained by mRNA analysis. However, in contrast to the mRNA results, GS52 protein was found in stems. Interestingly, anti-GS52 antibody recognized a 50-kDa protein found only in nodule extracts. Treatment of roots with anti-GS52 antibody, but not anti-GS50 antibody or preimmune serum, blocked nodulation by B. japonicum. Fractionation of cellular membranes in sucrose density gradients and subsequent Western analysis of the fractions revealed that GS50 colocalized with marker enzymes for the Golgi, while GS52 colocalized with marker enzymes for the plasma membrane. Restriction fragment length polymorphism (RFLP)-based mapping placed the gs52 gene on major linkage group J of the integrated genetic map of soybean. These data suggest that GS50 is likely an endo-apyrase involved in Golgi function, while GS52 is localized on the root surface and appears to play an important role in nodulation. PMID- 11043469 TI - A second T-region of the soybean-supervirulent chrysopine-type Ti plasmid pTiChry5, and construction of a fully disarmed vir helper plasmid. AB - Agrobacterium tumefaciens Chry5, which is particularly virulent on soybeans, induces tumors that produce a family of Amadori-type opines that includes deoxyfructosyl glutamine (Dfg) and its lactone, chrysopine (Chy). Cosmid clones mapping to the right of the known oncogenic T-region of pTiChry5 conferred Amadori opine production on tumors induced by the nopaline strain C58. Sequence analysis of DNA held in common among these cosmids identified two 25-bp, direct repeats flanking an 8.5-kb segment of pTiChry5. These probable border sequences are closely related to those of other known T-regions and define a second T region of pTiChry5, called T-right (TR), that confers production of the Amadoriopines. The oncogenic T-left region (TL) was located precisely by identifying and sequencing the likely border repeats defining this segment. The two T-regions are separated by approximately 15 kb of plasmid DNA. Based on these results, we predicted that pKYRT1, a vir helper plasmid derived from pTiChry5, still contains all of TR and the leftmost 9 kb of TL. Consistent with this hypothesis, transgenic Arabidopsis thaliana plants selected for with a marker encoded by a binary plasmid following transformation with KYRT1 co-inherited production of the Amadori opines at high frequency. All opine-positive transgenic plants also contained TR-DNA, while those plants that lacked TR-DNA failed to produce the opines. Moreover, A. thaliana infected with KYRT1 in which an nptII gene driven by the 35S promoter of Cauliflower mosaic virus was inserted directly into the vir helper plasmid yielded kanamycin-resistant transformants at a low but detectable frequency. These results demonstrate that pKYRT1 is not disarmed, and can transfer Ti plasmid DNA to plants. A new vir helper plasmid was constructed from pTiChry5 by two rounds of sacB-mediated selection for deletion events. This plasmid, called pKPSF2, lacks both of the known T-regions and their borders. pKPSF2 failed to transfer Ti plasmid DNA to plants, but mobilized the T region of a binary plasmid at an efficiency indistinguishable from those of pKYRT1 and the nopaline-type vir helper plasmid pMP90. PMID- 11043470 TI - A potato gene encoding a WRKY-like transcription factor is induced in interactions with Erwinia carotovora subsp. atroseptica and Phytophthora infestans and is coregulated with class I endochitinase expression. AB - A potato gene encoding a putative WRKY protein was isolated from a cDNA library enriched by suppression subtractive hybridization for sequences upregulated 1 h postinoculation with Erwinia carotovora subsp. atroseptica. The cDNA encodes a putative polypeptide of 172 amino acids, containing a single WRKY domain with a zinc finger motif and preceded by a potential nuclear localization site. St-WRKY1 was strongly upregulated in compatible, but only weakly in incompatible, interactions with Phytophthora infestans where, in all cases, it was coregulated with class I endochitinase, associating its expression with a known defense response. Whereas St-WRKY1 was strongly induced by E. carotovora culture filtrate (CF), confirming it to be an elicitor-induced gene, no such induction was detected after treatment with salicylic acid, methyl jasmonate, ethylene, or wounding. St-WRKY1 was upregulated by treatment of potato leaves with CFs from recombinant Escherichia coli containing plasmids expressing E. carotovora pectate lyase genes pelB and pelD, suggesting that either proteins encoded by these genes, or oligogalacturonides generated by their activity, elicit a potato defense pathway associated with St-WRKY1. PMID- 11043471 TI - The cylindrical inclusion gene of Turnip mosaic virus encodes a pathogenic determinant to the Brassica resistance gene TuRB01. AB - The viral component of Turnip mosaic virus (TuMV) determining virulence to the Brassica napus TuRB01 dominant resistance allele has been identified. Sequence comparisons of an infectious cDNA clone of the UK 1 isolate of TuMV (avirulent on TuRB01) and a spontaneous mutant capable of infecting plants possessing TuRB01 suggested that a single nucleotide change in the cylindrical inclusion (CI) protein coding region (gene) of the virus was responsible for the altered phenotype. A second spontaneous mutation involved a different change in the CI gene. The construction of chimeric genomes and subsequent inoculations to plant lines segregating for TuRB01 confirmed the involvement of the CI gene in this interaction. Site-directed mutagenesis of the viral coat protein (CP) gene at the ninth nucleotide was carried out to investigate its interaction with TuRB01. The identity of this nucleotide in the CP gene did not affect the outcome of the viral infection. Both mutations identified in the CI gene caused amino acid changes in the C terminal third of the protein, outside any of the conserved sequences reported to be associated with helicase or cell-to-cell transport activities. This is the first example of a potyvirus CI gene acting as a determinant for a genotype-specific resistance interaction. PMID- 11043472 TI - The Lotus japonicus LjSym4 gene is required for the successful symbiotic infection of root epidermal cells. AB - The role of the Lotus japonicus LjSym4 gene during the symbiotic interaction with Mesorhizobium loti and arbuscular mycorrhizal (AM) fungi was analyzed with two mutant alleles conferring phenotypes of different strength. Ljsym4-1 and Ljsym4-2 mutants do not form nodules with M. loti. Normal root hair curling and infection threads are not observed, while a nodC-dependent deformation of root hair tips indicates that nodulation factors are still perceived by Ljsym4 mutants. Fungal infection attempts on the mutants generally abort within the epidermis, but Ljsym4-1 mutants allow rare, successful, infection events, leading to delayed arbuscule formation. On roots of mutants homozygous for the Ljsym4-2 allele, arbuscule formation was never observed upon inoculation with either of the two AM fungi, Glomus intraradices or Gigaspora margarita. The strategy of epidermal penetration by G. margarita was identical for Ljsym4-2 mutants and the parental line, with appressoria, hyphae growing between two epidermal cells, penetration of epidermal cells through their anticlinal wall. These observations define a novel, genetically controlled step in AM colonization. Although rhizobia penetrate the tip of root hairs and AM fungi access an entry site near the base of epidermal cells, the LjSym4 gene is necessary for the appropriate response of this cell type to both microsymbionts. We propose that LjSym4 is required for the initiation or coordinated expression of the host plant cell's accommodation program, allowing the passage of both microsymbionts through the epidermis layer. PMID- 11043473 TI - Both induction and morphogenesis of cyst nematode feeding cells are mediated by auxin. AB - Various lines of evidence show that local changes in the auxin concentration are involved in the initiation and directional expansion of syncytia induced by cyst nematodes. Analysis of nematode infections on auxin-insensitive tomato and Arabidopsis mutants revealed various phenotypes ranging from complete inhibition of syncytium development to a decrease in hypertrophy and lateral root formation at the infection site. Specific activation of an auxin-responsive promoter confirmed the role of auxin and pointed at a local accumulation of auxin in developing syncytia Disturbance of auxin gradients by inhibiting polar auxin transport with N-(1-naphthyl)phtalamic acid (NPA) resulted in abnormal feeding cells, which were characterized by extreme galling, massive disordered cell divisions in the cortex, and absence of radial expansion of the syncytium initial toward the vascular bundle. The role of auxin gradients in guiding feeding cell morphogenesis and the cross-talk between auxin and ethylene resulting in a local activation of cell wall degrading enzymes are discussed. PMID- 11043474 TI - The broad-spectrum tospovirus resistance gene Sw-5 of tomato is a homolog of the root-knot nematode resistance gene Mi. AB - We used a positional cloning approach to isolate the Sw-5 disease resistance locus of tomato. Complementation experiments with overlapping cosmid clones enabled us to demonstrate that Sw-5 is a single gene locus capable of recognizing several tospovirus isolates and species. Analysis of the predicted Sw-5 protein suggests that it is a cytoplasmic protein, with a potential nucleotide binding site (NBS) domain and a C-terminal end consisting of leucine-rich repeats (LRRs). Based on its structural features, Sw-5 belongs to the class of NBS-LRR resistance genes that includes the tomato Mi, 12, and Prf genes; the Arabidopsis RPM1 gene; and the plant potato virus X resistance gene Rx. The overall similarity between the Sw-5 and Mi proteins of tomato suggests that a shared or comparable signal transduction pathway leads to both virus and nematode resistance in tomato. The similarity also supports the hypothesis that Sw-5 provides resistance via a hypersensitive response. Sw-5 is a member of a loosely clustered gene family in the telomeric region of chromosome 9. Members of this family map to other regions of chromosome 9 and also to chromosome 12, where several fungal, virus, and nematode genes have been mapped, suggesting that paralogs of Sw-5 may have evolved to provide different resistance specificities. PMID- 11043475 TI - Susceptibility and symptom development in Arabidopsis thaliana to Tobacco mosaic virus is influenced by virus cell-to-cell movement. AB - To identify host factors that regulate susceptibility to Tobacco mosaic virus (TMV), 14 Arabidopsis thaliana ecotypes were screened for their ability to support TMV systemic movement. The susceptibility phenotypes observed included one ecotype that permitted rapid TMV movement accompanied by symptoms, nine ecotypes that allowed a slower intermediate rate of systemic movement without symptoms, and four ecotypes that allowed little or no systemic TMV movement. Molecular comparisons between ecotypes representing the rapid (Shahdara), intermediate (Col-1), and slow (Tsu-1) movement phenotypes revealed a positive correlation between the ability of TMV to move cell to cell and its speed of systemic movement. Additionally, protoplasts prepared from all three ecotypes supported similar levels of TMV replication, indicating that viral replication did not account for differences in systemic movement. Furthermore, induction of the pathogenesis-related genes PR-1 and PR-5 occurred only in the highly susceptible ecotype Shahdara, demonstrating that reduced local and systemic movement in Col-1 and Tsu-1 was not due to the activation of known host defense responses. Genetic analysis of F2 progeny derived from crosses made between Shahdara and Tsu-1 or Col-1 and Tsu-1 showed the faster cell-to-cell movement phenotypes of Shahdara and Col-1 segregated as single dominant genes. In addition, the Shahdara symptom phenotype segregated independently as a single recessive gene. Taken together, these findings suggest that, within Arabidopsis ecotypes, at least two genes modulate susceptibility to TMV. PMID- 11043476 TI - Fructose utilization and phytopathogenicity of Spiroplasma citri. AB - Spiroplasma citri is a plant-pathogenic mollicute. Recently, the so-called nonphytopathogenic S. citri mutant GMT 553 was obtained by insertion of transposon Tn4001 into the first gene of the fructose operon. Additional fructose operon mutants were produced either by gene disruption or selection of spontaneous xylitol-resistant strains. The behavior of these spiroplasma mutants in the periwinkle plants has been studied. Plants infected via leafhoppers with the wild-type strain GII-3 began to show symptoms during the first week following the insect-transmission period, and the symptoms rapidly became severe. With the fructose operon mutants, symptoms appeared only during the fourth week and remained mild, except when reversion to a fructose+ phenotype occurred. In this case, the fructose+ revertants quickly overtook the fructose- mutants and the symptoms soon became severe. When mutant GMT 553 was complemented with the fructose operon genes that restore fructose utilization, severe pathogenicity, similar to that of the wild-type strain, was also restored. Finally, plants infected with the wild-type strain and grown at 23 degrees C instead of 30 degrees C showed late symptoms, but these rapidly became severe. These results are discussed in light of the role of fructose in plants. Fructose utilization by the spiroplasmas could impair sucrose loading into the sieve tubes by the companion cells and result in accumulation of carbohydrates in source leaves and depletion of carbon sources in sink tissues. PMID- 11043477 TI - Structural and expression analysis of uricase mRNA from Lotus japonicus. AB - Uricase (nodulin-35) cDNA, LjUr, was isolated from nodules of a model legume, Lotus japonicus. LjUr expression was most abundant in nodules, although it was detected in nonsymbiotic tissues as well, particularly in roots. Expression in nodules was detected in uninfected cells, nodule parenchyma, and, more intensely, in vascular bundles. Phylogenetic analysis of uricase sequences from various legumes indicated that uricases of amide- and ureide-transporting legumes form two distinct clades. LjUr is in the cluster of amide-transport legumes even though L. japonicus bears determinate nodules. PMID- 11043478 TI - Greetings to nurses from Her Majesty Empress Michiko of Japan. PMID- 11043479 TI - Sharing knowledge to create an uplifting future. PMID- 11043480 TI - Nurses, physicians and pharmacists create a global alliance. PMID- 11043481 TI - Nurses remind G8 leaders that poverty impacts on health. PMID- 11043482 TI - ICN's Code of Ethics for Nurses: Serving nurses and nursing care world-wide. PMID- 11043483 TI - The International Council of Nurses Code of Ethics for Nurses. PMID- 11043484 TI - From vision to reality: how to actualize the vision of discharging patients from a hospital, with an increased focus on prevention. AB - Experiences in creating dialogue possibilities to stimulate interdisciplinary and intersectoral collaboration in hospital discharge and prevention are presented. Time is often a major constraint that persuades decision-makers to avoid using qualitative methodologies in research and development. Quick results are demanded of today's health-care system, not allowing ideas to be implemented or visions owned by professionals. Action-orientated research is used and recommended, despite its time-demanding methodology. A multidisciplinary management team and the authors, over a 2-year period, worked with the development and implementation of 'Preventive Discharge' in a Danish hospital clinic. The empirical starting point, developmental processes, piloting and implementation of the project are described. Earlier research from a literature review on discharge is referenced. The results show that while there are many barriers to dialogue between professions and between organizational levels, it is possible to frame such opportunities and improve interdisciplinary and intersectoral collaboration for health. External supervision may be an effective tool for stimulating dialogue. The testing phase afforded a valuable lesson when pilot-site collaborators, initially not involved in the development of the process tools, reformulated the project to suit their cultural climate while maintaining the project's original aims. Finally, a discussion is presented on actualizing the vision for 'Preventive Discharge'. PMID- 11043485 TI - The older adult: a comparison of long-term care in Glasgow and San Francisco. AB - Population trends show an increasing proportion of people older than 65 years of age. This report aims to describe the facilities/services accessible to the older adult living in Glasgow, Scotland and in San Francisco, United States of America. This comparative study serves to develop a broader understanding of what is involved in the process of ageing, and describes how these two cities are responding to the needs of older people. Long-term caring provision is complex and involves multiple agencies, often leaving both lay and professional individuals ill-informed as to which services exist and how best to use them. The nurse can be instrumental in providing information, viable alternatives and services. In this article, options available for caring of the older adult and financing of care are discussed along with a number of recommendations based on observations described, data collected and literature researched. With the advent of care being provided in the community, there is now a continuum of services ranging from independent living at home to dependent care in hospital. Whether an individual remains at home depends on the availability and affordability of, and access to, other resources/facilities. Creative alternatives are needed for care of the older adult in this millennium. This report describes the need for an integrated system of care, which can adjust to variations in clients' needs. Progressive care facilities, intergenerational sharing, community volunteer service programmes for the older person and Edenizing are a few recommendations discussed by the authors. PMID- 11043486 TI - Meeting patients' skin care needs: harnessing nursing expertise at an international level. AB - Skin disease is highly prevalent, particularly in the developing world. This can result in skin failure which in turn may have a major social and economic impact on individuals and communities (Finlay & Ryan 1996). The significant lack of dermatologists worldwide, most of whom are based in hospitals, means that expertise in skin care cannot always be delivered to those who need it. This paper advances the argument that a care delivery model is needed which provides skin care focused at a primary care level throughout the nursing service, drawing on specialists who are often based in the secondary health care facilities. This can only be achieved through adopting a strategic approach which identifies the training needs of such professionals, harnesses the appropriate expertise, shares good practice, and operates in close conjunction with dermatologists. This paper outlines how this vision can be put into operation, by outlining the strategic international development work being undertaken by nurses and the efforts to promote inter-professional collaboration. PMID- 11043487 TI - Regional Examination for Nurse Registration, Commonwealth Caribbean. AB - In 1990, a Regional Examination for Nurse Registration was approved by the Conference of Ministers Responsible for Health in the Commonwealth Caribbean. The examination allows for standardization and improvement of nursing education, as well as reciprocity and ease-of-movement for Registered Nurses among the countries of the region. A Planning Committee and a Blueprint Committee were established to develop the examination process. Committee membership included the Principal/Chief Nursing Officers, Nurse Tutors, and General Nursing Council representatives of each country, as well as educators from the two universities of the region. The accepted model, based on mutually agreed competencies for the Registered Nurse to practice in the region, forms the basis for the elaboration of the blueprint and the administrative manuals. The treatment of test items, assembling and conducting the examinations, which consist of four papers, as well as scoring the examination results, and notifying students of the results, are the responsibility of each General Nursing Council. The 13 General Nursing Councils with responsibility for Schools of Nursing meet annually as a regional committee to prepare the examinations, using a different country for each meeting; countries are rotated alphabetically. There is no permanent site for the administration of the examination. Membership to the regional committee, known as the Regional General Nursing Councils (RGNCs), is for 3 years. This staggering of membership allows for continuity. PMID- 11043488 TI - A client-centered model: discharge planning in Juvenile Justice Centres in New South Wales, Australia. AB - Health care delivered in Juvenile Justice Centres in New South Wales, Australia, leads to improvement in the health status of detainees while in custody but this declines on discharge and is contributed to by a breakdown in continuity of care. In an effort to halt this decline in health status, a model of interdisciplinary discharge planning is proposed to address the interface between Juvenile Justice Centres and adult correctional facilities and the community. Key features of the process are: collaboration through the development of links by establishment of a community liaison co-ordinator position; open and continuous communication between service providers; continuous involvement of detainees and their significant others; and ongoing support to detainees once discharged. Evaluation of the discharge-planning process ensures that the process is appropriate to meet the needs of this specific and diverse population of young people and supplies feedback to all service providers. The trusting relationship that is developed between detainees and registered nurses within centres leaves nurses ideally placed to co-ordinate the discharge-planning process. PMID- 11043489 TI - Bismuth biokinetics and kidney histopathology after bismuth overdose in rats. AB - Bismuth induced nephrotoxicity has been reported to occur after acute overdoses of Bi-containing therapeutic drugs. We studied the development of bismuth induced nephropathy and bismuth biokinetics in rats. Bismuth nephropathy was induced in 33 young adult female Wistar rats weighing ca. 175 g by feeding them a single overdose of colloidal bismuth subcitrate containing 3.0 mmol Bi/kg at (t = 0). Control animals (n = 7) were fed the vehicle only. The animals were sacrificed after 1-48 h. Plasma creatinine increased from 51 +/- 6 micromol/l at t = 0 to 550 +/- 250 micromol/l after 48 h in the experimental group. The S3 segment of the proximal tubule showed epithelial cell vacuolation after 1 h and necrosis after 3 h. Cells of the S1/S2 segment demonstrated vacuolation after 6 h and necrosis after 12 h. Biokinetics of bismuth in blood could best be described with a one-compartment model characterized by an absorption half-life of 0.32 h and an elimination halflife of 16 h. The peak concentration of about 7.0 mg Bi/l was reached after 2 h. In conclusion, cells of the S3 segment of the proximal tubule necrotized first after an oral colloidal bismuth subcitrate overdose and biokinetics of Bi in blood was best described by a one-compartment model. PMID- 11043490 TI - Percutaneous absorption and metabolism of dinitrochlorobenzene in vitro. AB - Dinitrochlorobenzene (DNCB) absorption through mouse and rat dorsal skin, pig ear skin and human abdominal skin in vitro was determined, and local metabolism to the glutathione conjugate was related to glutathione transferase activities and glutathione status in the skin. Absorption studies were conducted using skin mounted in a flow-through diffusion cell with tissue culture medium as receptor fluid. DNCB applied to the surface of skin in acetone penetrated through 26-day old rat skin better than through the skin of the other species investigated. The amounts of absorption through pig and human skin and conjugation formation were similar. In general, occlusion resulted in increased penetration of DNCB but no change in conjugation. Human skin showed the highest gluta-thione-S-transferase activity towards DNCB, followed by 26-day-old rat, pig, mouse and neonatal rat skin. Levels of glutathione were highest in mouse skin, followed by neonatal rat, 26-day-old rat, pig and human skin, with pig and human skin showing similar levels. These studies indicated that the glutathione level in skin was the determining factor influencing the degree of DNCB conjugation during percutaneous absorption, and this was greatly depleted during percutaneous penetration of DNCB. PMID- 11043491 TI - Influence of subchronic administration of oestrone-3-O-sulphamate on oestrone sulphatase activity in liver, spleen and white blood cells of ovariectomized rats. AB - Inhibition of oestrone sulphatase followed by oestrogen removal from tumour cells may be a new form of endocrine therapy of breast cancer in women. We investigated the inhibitory effect of the subchronic administration of oestrone-3-O-sulphamate (EMATE), a steroid sulphatase inhibitor, to ovariectomized rats, to evaluate this method for testing new nonsteroidal inhibitors. EMATE in DMSO was administered both orally and subcutaneously (s.c.) for 7 days at doses of 0.5 and 2.5 mg/kg. In addition the rats were injected s.c. with 0.5 mg oestrone sulphate/kg 26 and 2 h before decapitation under ether anaesthesia. Oestrone sulphatase activity (ESA) was measured radiometrically using [3H]oestrone sulphate as substrate for desulphuration in white blood cells, liver homogenate, microsomes and spleen homogenate. ESA in liver microsomes was found to be nearly 40 times higher than in white blood cells while in spleen ESA was nearly half of that found in liver homogenates and white blood cells. ESA can be inhibited by EMATE down to 50-1.5% of control activity depending on the dose and administration route. The inhibition was in the order, liver homogenate < spleen < liver microsomes < white blood cells, and was more pronounced after s.c. administration of the inhibitor than after oral administration. Ovariectomy was found to be not necessary for oestrone sulphatase-inhibiting studies. Two sequential s.c. injections of oestrone sulphate enhanced the enzyme activities significantly in liver and white blood cells, but not in spleen. In conclusion, white blood cells and liver microsomes of intact female rats can be used for ESA-inhibiting studies. Sulphate conjugated oestrone can induce oestrone sulphatase in vivo in liver and white blood cells thereby enhancing oestrogen supply in the peripheral organs. PMID- 11043492 TI - Characterization of liver microsomal 7-ethoxycoumarin O-deethylation and chlorzoxazone 6-hydroxylation activities in Japanese and Caucasian subjects genotyped for CYP2E1 gene. AB - To determine whether the CYP2E1 genetic polymorphisms cause alterations in protein expression and enzyme catalytic activities, three CYP2E1 genetic polymorphisms, namely RsaI/PstI, DraI, and MspI types, were determined in liver genomic DNA isolated from 39 Japanese and 45 Caucasians. These genotypes were compared with levels of CYP2E1 and activities of 7-ethoxycoumarin O-deethylation and chlorzoxazone 6-hydroxylation in liver microsomes from these human samples. In combination of three types of CYP2E1 polymorphisms, it was classified into seven genotypes in the Japanese population and four in the Caucasian population. The incidence in the occurrence of RsaI/PstI polymorphism or DraI polymorphism was 0.24 and 0.29 for Japanese, and 0.01 and 0.02 for Caucasians. Ethnic difference was also noted in the MspI polymorphism in which frequencies in Japanese and Caucasian populations were 0.15 and 0.02, respectively. Studies with liver microsomes showed that there were no significant differences in the levels of expression of CYP2E1 protein between wild-type (group A) and other 6 genotypes (B, C, D, E, F, and G) in Japanese and other three genotypes (B, D, and F) in Caucasians. Catalytic activities for 7-ethoxycoumarin O-deethylation and chlorzoxazone 6-hydroxylation by liver microsomes were also found to be less significantly affected by mutations in the CYP2E1 gene in human samples examined in this study. These results support the view that RsaI/PstI, DraI, and MspI types of CYP2E1 genetic polymorphisms may not cause significant alterations in protein expression and enzyme catalytic activities of CYP2E1 enzyme in human livers. PMID- 11043493 TI - Establishment of a novel in vitro system for studying the interaction of xenobiotic metabolism of liver and intestinal microflora. AB - We developed a new two-chamber system for the coculture of hepatocytes and fecal microflora under aerobic and anaerobic conditions, respectively, to investigate the sequential metabolism of chemicals by the liver and microflora in vitro. The culture device consisted of two chambers separated by a permeable polycarbonate membrane. In the aerobic compartment, hepatocytes were cultivated as a monolayer on the membrane and in the anaerobic compartment fecal microflora as a suspension. To characterize the metabolic capacity of the microflora and hepatocytes, various marker enzymes were studied. Azoreductase, nitroductase, beta-glucuronidase, beta-glucosidase and sulphatase were tested in the microflora of the feces from three volunteers who had had significantly different eating habits for years (daily meat, mixed diet, vegetarian). The microflora exhibited significant activities and the various enzymes differed only moderately in the samples from the three volunteers. For rat hepatocytes the activities of various cytochrome P450 forms and conjugating enzymes served as markers. The enzyme activities were tested in the coculture system during a 4-h culture period intended for the test protocol. Deethylation of ethoxycoumarin and 2alpha-, 6beta and 16alpha-hydroxylation of testosterone decreased by about 30%, 25%, 40% and 20%, respectively, while there was no loss of glucuronidation and sulphonation of 3-OH-benzo(a)pyrene nor of glutathione conjugation of 1-chloro-2,4-dinitrobenzene during the 4-h culture period. The activities of the tested hepatic phase I and II enzymes were not changed after coculture of the hepatocytes with the microflora for 4 h. The applicability of the in vitro system for studying the metabolic interaction of liver and microflora was demonstrated using 7 ethoxycoumarin and the developmental drug EMD 57033, a thiadiazinon derivative from Merck KGaA, as model compounds. Both compounds were oxidized and conjugated by liver cells. In the coculture of hepatocytes and fecal microflora the resulting glucuronides and sulphoconjugates were split by hydrolytic enzymes of the intestinal microflora. PMID- 11043494 TI - Placental transfer and pharmacokinetics of a single oral dose of [14C]p nitrophenol in rats. AB - The pharmacokinetics and placental transfer of a single oral dose of 100 mg/kg (10 microCi/kg, 16% of acute oral LD50) of uniformly phenyl-labeled [14C]p nitrophenol were investigated in pregnant Sprague-Dawley rats at 14-18 days of gestation. Three animals were killed on gestation day 18, at 0.5, 1, 2, 4, 12, 24, and 48 h after dosing. Radioactivity was rapidly absorbed and distributed throughout the maternal and fetal tissues. The gastrointestinal tract contents retained 20% and 2% of the dose at 0.5 h and 4 h after dosing. The peak maternal plasma concentration of radioactivity (microg p-nitrophenol equivalent/ml) was 7.17 compared with 0.37 for fetal plasma at 0.5 h. Maximum concentration of radioactivity (microg p-nitrophenol equivalent/g fresh tissue) was detected in most tissues 0.5 h after dosing and was in descending order: kidney 23.27, liver 12.37, placenta 3.56, fetus 2.17, and brain 1.99. Radioactivity was eliminated from plasma and all tissues beiexponentially. The half-lives of elimination of 14C were 34.65 h and 69.30 h for maternal and fetal plasma, respectively. p Nitrophenol, detected by HPLC, was the major compound identified in plasma and tissues. While p-nitrophenol disappeared biphasically from maternal plasma and kidney, it was eliminated monophasically from brain, placenta, and liver. p Nitrocatechol and p-aminophenol were detected in the liver with peak concentrations at 0.5 h of 1.13 and 1.00 microg/g fresh tissue, respectively. While the change in the concentration of p-nitrocatechol with time was monophasic, that of p-aminophenol showed a biphasic pattern with elimination half lives of 1.93 h and 4.95 h, respectively. Radioactivity was rapidly excreted in the urine mostly as polar metabolites, while only 3% of the dose was recovered in the feces. Radioactive materials excreted in the urine comprised: glucuronides 4%, sulfates 8%, hot-acid hydrolysates 11%, nonconjugated compounds 16%, and water-soluble metabolites 61%. This study demonstrated that although orally administered p-nitrophenol is a rapidly absorbed and excreted compound, it is transported to the maternal brain and the fetus and may pose a health risk following exposure to toxic doses during pregnancy. PMID- 11043495 TI - Inhalation of toluene diisocyanate affects cytochrome P450 2B1 expression in rat lung. AB - In the lung the expression of xenobiotic-metabolizing enzymes such as cytochromes P450 (CYP) and glutathione S-transferases (GST) may be affected by inhaled pollutants. Toluene diisocyanate (TDI) is a highly volatile chemical compound known to induce a wide array of diseases in workers exposed to vapors or sprays, including respiratory allergy and asthma. We investigated the effect of inhaled TDI on expression of CYP 1A1, 2B1, 2E1, and 3A1 and of alpha-, mu-, and pi-GST in rat lung. Animals were exposed to targeted concentrations of 0.01, 0.1, or 1 ppm TDI vapors or to cleaned filtered air for 8 h. Expression of CYP and GST was analyzed 18 24 h after the end of exposure using western blotting, northern blotting, and immunohistochemistry. Constitutive levels of CYP 2B1 and 3A1 proteins were found in lung tissue from control rats, whereas CYP 1A1 and 2E1 proteins were not detectable. Animal exposure to TDI vapors neither modified CYP 3A1 protein expression, nor led to any detectable expression of CYP 1A1 or 2E1. In contrast, exposure to 1 ppm TDI induced a 40% reduction in CYP 2B1 protein levels. This decrease was associated with a 33% decrease in CYP 2B1 mRNA levels. Additionally, CYP 2B1 immunolabeling localized to ciliated epithelial cells, Clara cells, and type II alveolar cells in the lung tissue of control rats was markedly decreased in animals exposed to 1 ppm TDI. Constitutive levels of alpha , mu-, and pi-GST proteins were found in lung tissue from control rats. Exposure to TDI had no effect on lung expression of either of the GST. In conclusion, this study clearly shows a selective decrease in CYP 2B1 expression by TDI vapors in rat lung. The contribution of CYP 2B1 to metabolize further xenobiotics is therefore altered. PMID- 11043496 TI - Development of a 32P-postlabelling method for the detection of 1,N2 propanodeoxyguanosine adducts of crotonaldehyde in vivo. AB - Crotonaldehyde is a genotoxic, mutagenic and carcinogenic alpha,beta-unsaturated carbonyl compound which forms 1,N2-propanodeoxyguanosine adducts. Humans are exposed to this compound at work places, and from tobacco smoke and air pollution, but also from food and beverages. Therefore crotonaldehyde can play a significant role in carcinogenesis. Since in vivo measurement of DNA adducts of crotonaldehyde can improve cancer risk assessment and contribute to the clarification of the role of crotonaldehyde in carcinogenicity, we developed, adapted and optimized a 32P-postlabelling technique for the adducts of crotonaldehyde based on nuclease P1 enrichment and on a polyethylene imine modified cellulose TLC to provide a detection sensitivity of three adducts per 10(9) nucleotides and a labelling efficiency of 80-90%. We also report a readily performable synthesis of adduct standards and demonstrated that DNA is completely digested to the 3'-monophosphate nucleotides under the conditions of our enzymatic DNA hydrolysis. We showed that the postlabelling method developed is appropriate for in vivo DNA-binding studies. Female Fischer 344 rats were treated by gavage with crotonaldehyde at doses of 200 and 300 mg/kg body weight, and 20 h after treatment adduct levels of 2.9 and 3.4 adducts per 10(8) nucleotides, respectively, were found in the liver DNA. Only 1.6 nucleotides per 10(8) nucleotides were found 12 h after treatment at 200 mg/kg body weight. Absolutely no adducts could be found in liver DNA of untreated rats with our method at the detection limit of three adducts per 10(9) nucleotides. In contrast to our group, the group of Chung have reported crotonaldehyde adduct levels in the range of 2.2 22 adducts per 10(8) nucleotides in DNA of untreated Fischer 344 rats. The clarification of this discrepancy is of importance for the elucidation of the role of crotonaldehyde in carcinogenicity, and both groups have decided to clarify this in cooperation in the near future. PMID- 11043497 TI - Genotoxic and mono-oxygenase system effects of the fungicide maneb. AB - The in vivo effects of a commercial preparation of maneb on mono-oxygenase activities of hepatic microsomes of basal and induced rats were examined. In vitro experiments with the D7 strain of yeast Saccharomyces cerevisiae were also performed. In both basal and induced rats maneb caused a decrease in cytochrome P 450 content and aniline hydroxylase. Immunoblotting analysis using anti-P-450 IIE1 antibodies confirmed the data obtained for aniline hydroxylase activity. Maneb was toxic in cells of S. cerevisiae. On the basis of in vivo and in vitro experiments it can be concluded that maneb possesses a toxic activity attributable to its main metabolite ethylene thiourea. Immunoblotting analysis indicates that maneb biotransformation influences the IIEI P-450 isoform. PMID- 11043498 TI - Stress and molecular chaperones in disease. AB - Stress, a common phenomenon in today's society, is suspected of playing a role in the development of disease. Stressors of various types, psychological, physical, and biological, abound. They occur in the working and social environments, in air, soil, water, food, and medicines. Stressors impact on cells directly or indirectly, cause protein denaturation, and elicit a stress response. This is mediated by stress (heat-shock) genes and proteins, among which are those named molecular chaperones because they assist other proteins to achieve and maintain a functional shape (the native configuration), and to recover it when partially lost due to stress. Denatured proteins tend to aggregate and precipitate. The same occurs with abnormal proteins due to mutations, or to failure of post transcriptional or post-translational mechanisms. These abnormal proteins need the help of molecular chaperones as much as denatured molecules do, especially during stress. A cell with normal antistress mechanisms, including a complete and functional set of chaperones, may be able to withstand stress if its intensity is not beyond that which will cause irreversible protein damage. There is a certain threshold that normal cells have above which they cannot cope with stress. A cell with an abnormal protein that has an intrinsic tendency to misfold and aggregate is more vulnerable to stress than normal counterparts. Furthermore, these abnormal proteins may precipitate even in the absence of stress and cause diseases named proteinopathies. It is possible that stress contributes to the pathogenesis of proteinopathies by promoting protein aggregation, even in cells that possess a normal chaperoning system. Examples of proteinopathies are age related degenerative disorders with protein deposits in various tissues, most importantly in the brain where the deposits are associated with neuronal degeneration. It is conceivable that stress enhances the progression of these diseases by facilitating protein unfolding and misfolding, which lead to aggregation and deposition. A number of reports in the last few years have described research aimed at elucidating the role of heatshock proteins, molecular chaperones in particular, in the pathogenesis of neurodegenerative disorders. The findings begin to shed light on the molecular mechanism of protein aggregation and deposition, and of the ensuing cell death. The results also begin to elucidate the role of molecular chaperones in pathogenesis. This is a fascinating area of research with great clinical implications. Although there are already several experimental models for the study of proteinopathies, others should be developed using organisms that are better known now than only a few years ago and that offer unique advantages. Use of these systems and of information available in databases from genome sequencing efforts should boost research in this field. It should be possible in the not-too-distant future to develop therapeutic and preventive means for proteinopathies based on the use of heat-shock protein and molecular chaperone genes and proteins. PMID- 11043499 TI - Cyclosporine-A plus steroids versus steroids alone in the 12-month treatment of systemic lupus erythematosus. AB - The positive results obtained with cyclosporine-A both in an experimental model and in selected patients with advanced systemic lupus erythematosus support the hypothesis that the drug could be used as a steroid sparer in the earliest stages of active disease. To determine the 12-month clinical efficacy (disease control and steroid sparing), safety, and tolerability of low-dose cyclosporine-A plus steroids versus steroids alone, we designed a multicenter, open, prospective, randomized, pilot study, controlled for parallel groups. The patients were then followed up to month 24. A total of 18 consenting patients with recently diagnosed systemic lupus erythematosus of moderate severity indicated for the use of steroids in acute boluses and subsequently per os were enrolled at two university hospital medical centers. The protocol was based on three 1-g boluses of 6-methylprednisolone followed by cyclosporine-A (<5 mg/kg per day) plus prednisone 0.5-1 mg/kg per day per os, reduced by 5 mg/day every 2 weeks following clinical remission, versus the same doses of oral prednisone alone. The efficacy evaluation was based on a four-point scale (from absent/none to severe) for signs and symptoms of systemic lupus erythematosus and immunoserological parameters. The disease activity index and cumulative prednisone dose per patient were analyzed. Any adverse events were reported. All patients showed a reduction in disease activity index within the 1st month. The results were significantly better in the group with cyclosporine-A plus prednisone throughout month 12 (baseline and 12-month disease activity indexes: 21.3+/-8.6 and 5.0+/-2.5 versus 20.4+/-7.1 and 8.8+/-6.0 in the prednisone group, P<0.05). The 12-month cumulative mean dose of prednisone was significantly lower in the group with both cyclosporine-A plus prednisone (179.4+/-40.1 versus 231.8+/-97.1 mg/kg, P<0.005). No unusual adverse events related to the study drugs have been reported. In particular, renal function and blood pressure monitoring revealed no significant changes from mean baseline values in either group. No disease flares were reported in the group treated with cyclosporine-A plus prednisone during the 12- to 24-month period. Thus cyclosporine-A represents a useful corticosteroid sparer in the maintenance of clinical remission in patients with an early-stage, active systemic lupus erythematosus. PMID- 11043500 TI - Measurement of soluble adhesion molecules in primary Raynaud's phenomenon and in Raynaud's phenomenon secondary to connective tissue diseases. AB - Adhesion molecules play a role in the inflammation and pathogenesis of vascular diseases. In 13 patients with primary Raynaud's phenomenon, 19 with Raynaud's phenomenon associated with connective tissue disease, and 16 control subjects, we measured plasma levels of soluble forms of intercellular adhesion molecule-1, vascular cell adhesion molecule-1, E-selectin, and von Willebrand factor. Patients with secondary Raynaud's phenomenon had plasma levels of soluble forms of intercellular adhesion molecule- 1, vascular cell adhesion molecule-1, E selectin, and von Willebrand factor which were significantly higher than in those with primary Raynaud's phenomenon and controls, while no difference was observed between patients with primary Raynaud's phenomenon and controls. Within the group with secondary Raynaud's phenomenon, the strongest correlations were between soluble forms of intercellular adhesion molecule-1 and both E-selectin, (r=0.67, P<0.001) and von Willebrand factor (r=0.58, P<0.01). In none of the three groups were the levels of soluble adhesion molecules and von Willebrand factor changed by exposure of hands to cold, although all patients had a definite vasospasm. In conclusion, this study indicates that primary Raynaud's phenomenon is not associated with elevation of soluble adhesion molecules and von Willebrand factor. Prospective studies are now required to investigate the role of these molecules as predictors of secondary diseases. PMID- 11043501 TI - Age-associated changes in nitric oxide metabolites nitrite and nitrate. AB - Aging is an important determinant of vascular disease. Endothelial dysfunction accompanying vascular disease may be related to cardiovascular risk factors such as aging, hypertension, and atherosclerosis. Experimental models suggest that endothelium-derived nitric oxide is reduced with aging, and this reduction is implicated in atherogenesis. The aim of this study was to determine whether increased age resulted in altered serum nitrite and nitrate levels, end-products of nitric oxide, in healthy subjects. Sixty-nine healthy individuals were divided into five different age groups: group I (6-15 years), group II (16-30 years), group III (31-45 years), group IV (46-60 years), and group V (>61 years). In these subjects, serum nitrite was measured by the Griess reaction and nitrate by the nitrate reductase method. Statistical analysis showed that serum nitrite levels were not significantly different in any of the groups, while serum nitrate concentrations exhibited significant differences (P<0.001). These findings suggest that nitric oxide synthesis and/or secretion is reduced with age and consequently endothelium-dependent vasodilation is impaired. PMID- 11043502 TI - Beta2-microglobulin gene mutation is not a common mechanism of HLA class I total loss in human tumors. AB - One hundred and sixty-two tumor samples were analyzed for HLA class I expression using immunohistological techniques. HLA class I total loss (phenotype no. I) was detected in 31 cases (19%), comprising 20 colorectal, 3 laryngeal, and 2 bladder carcinomas and 6 melanomas. Twenty-one cases were selected for molecular analysis due to a higher proportion of tumor cells versus stroma cells (75%). We investigated whether beta2-microglobulin mutation was responsible for HLA downregulation. Single-strand conformation polymorphism and sequencing analysis of DNA samples was performed. Alterations were detected only in melanomas M78 (a point mutation in the initiation ATG sequence), M79 (a mutation in codon 31 producing a stop codon), and M34 (a TTCT deletion introducing a termination codon signal). We found no beta2-microglobulin gene mutation in the other 18 samples. Loss of heterozygosity in 15q close to the beta2-microglobulin gene was found in 5 cases. We conclude that HLA class I total loss can frequently occur without beta2-microglobulin gene mutations. PMID- 11043503 TI - Association of lipoprotein(a) concentration and apo(a) isoform size with restenosis after percutaneous transluminal coronary angioplasty. AB - Lp(a) is a unique class of lipoprotein particles that exhibits a considerable size heterogeneity resulting from the size polymorphism of apo(a), its unique protein component. An elevated level of Lp(a) in plasma has been proposed to be a risk factor for premature development of coronary artery disease. To evaluate the relationship between Lp(a) concentration and apo(a) isoform size with restenosis after percutaneous transluminal coronary angioplasty, Lp(a) levels and apo(a) phenotypes were determined in 204 patients who underwent a successful coronary angioplasty procedure and stent implantation. The patients were followed with clinical examinations and exercise tests at 1, 3, and 6 months, and a control coronary angiography was performed after 6 months to evaluate restenosis. Lp(a) levels were determined with an ELISA that is insensitive to the size heterogeneity of Lp(a), and the apo(a) isoforms were determined by a high resolution agarose gel electrophoresis method followed by immunoblotting with a specific monoclonal antibody. Of the 146 patients who underwent angiographic evaluation, 57 (39%) had restenosis, whereas 89 (61%) did not. Lp(a) levels and the distribution of the expressed apo(a) phenotypes were compared in these two groups of patients. Although the mean and median Lp(a) levels were higher in the restenosed group, the difference was not statistically significant. However, a significant difference in Lp(a) values was found in women (P=0.043), even though, because of the small number of women in the study (n=35), no sound conclusions can be reached on the predictive role of Lp(a) in restenosis. There also was no difference in the distribution of apo(a) phenotypes between the two groups. Because of their wide distribution, Lp(a) values and apo(a) isoforms do not seem to be a useful indicator of risk of restenosis after percutaneous transluminal coronary angioplasty in our study cohort. PMID- 11043505 TI - Lack of association between antiphospholipid antibodies and migraine in children. AB - Anticardiolipin antibodies are found frequently in those suffering from migraine, but it is not clear if this association is real or coincidental. Moreover, there are no data on the prevalence of anticardiolipin antibodies in children. In this study, 40 patients were divided into two groups according to the type of migraine: group I included 22 cases (15 females and 7 males, mean age+/-SD 13.7+/ 8.9 years) suffering from migraine with and without aura; group II consisted of 18 children (10 females and 8 males, age 14.7+/-6.9 years) having migraine with prolonged aura or migrainous infarction, also called complicated migraine. We studied two groups of children as controls: a group of 35 children (25 females and 10 males, mean age 13.9+/-7.1 years) with juvenile chronic arthritis (group III) and a group of 40 healthy sex- and age-matched children who did not suffer from migraine or any other neurological disease (group IV). No statistically significant differences in levels of anticardiolipin antibodies were found between group I and II and controls. Our data demonstrate that, in children with migraine, anticardiolipin antibodies are not more frequent than in healthy controls, and suggest that anticardiolipin antibodies are not implicated in the pathogenesis of migraine. PMID- 11043504 TI - Effects of defibrotide on aorta and brain malondialdehyde and antioxidants in cholesterol-induced atherosclerotic rabbits. AB - The effects of a high-cholesterol diet in the presence and absence of defibrotide, a single-stranded polydeoxyribonucleotide compound, on the lipid peroxidation product malondialdehyde, endogenous antioxidant enzymes catalase, glutathione peroxidase, and the antioxidant thiol compound GSH were investigated. Forty male New Zeland white rabbits were divided into four groups each consisting of 10 rabbits. Group I received a regular rabbit chow diet and group II 1% cholesterol plus regular chow, group III was given defibrotide (60 mg/kg per day p.o. in water) and was fed with regular chow, and group IV received defibrotide plus 1% cholesterol for 9 weeks. Blood cholesterol and malondialdehyde, catalase, glutathione peroxidase, and GSH were determined before starting the experimental diet regimen (basal). After 9 weeks, the same parameters were determined in blood, aorta, and brain tissues (end -experiment). Aortic tissue was examined under a light microscope for morphological alterations indicative of atherosclerosis. The increase in serum total cholesterol was greater in group II than group IV. Plasma malondialdehyde in group II was higher than in group III. Brain malondialdehyde in group II was higher than all other groups, and aortic malondialdehyde in this group was higher than group I and III. Serum catalase activity decreased in group II and increased in group III, compared with basal values. Brain catalase activity in group I was higher than group II, and aorta catalase in group IV was higher than in group I and III. Blood glutathione peroxidase activity in group III and IV was higher than basal. GSH concentrations decreased significantly in the cholesterol-fed groups (group II and IV). Histological alterations in the cholesterol-fed groups were more pronounced in group II. The increased levels of malondialdehyde in plasma, aorta, and brain tissue of group II suggest a role of oxygen free radicals in the pathogenesis of cholesterol-induced atherosclerosis. The higher malondialdehyde values in the brain tissues of animals in group II compared with group IV suggest a protective role of defibrotide in the brain against lipid peroxidation in the oxidant stress of cholesterol-induced atherosclerosis. Increased catalase activities in the blood and aortic tissues and increased glutathione peroxidase activities in the blood of rabbits receiving defibrotide suggest an induction of these antioxidant enzyme activities by defibrotide. These results imply that anti-atherosclerotic, anti-ischemic effects of this drug may be due to the beneficial effects on the oxidant-antioxidant balance of various tissues. PMID- 11043506 TI - A co-author dissociates himself from erroneous published results. PMID- 11043507 TI - Distribution of 32 alelle of the CCR5 gene in the population of Poland. AB - The chemokine receptor CCR5 constitutes a major co-receptor for the R5 strains of HIV-1, and a mutant allele of the CCR5 gene, especially in the homozygous form delta32/delta32, confers resistance against infection by the virus. The frequency of the delta32 allele was determined in blood donors from 16 provinces, covering the entire territory of Poland. Among 861 individuals 182 (21.1%) were carriers of the mutated allele: 7 of them (0.8 %) were homozygotes delta32/delta32, and 175 (20.3%) were heterozygotes +/delta32, resulting in a 10.9% frequency of the delta32 allele. The highest frequencies of the mutated allele were found in the eastern and western provinces, and the lowest frequencies of the delta32 allele were detected in the provinces in the center of the country. This pattern of distribution may reflect the migration of the population from the eastern territories of Poland to the western part of the country after World War II. PMID- 11043508 TI - Molecular analysis of Wilson disease in Taiwan: identification of one novel mutation and evidence of haplotype-mutation association. AB - Wilson disease (WND) is caused by a deficiency of the copper-transporting enzyme, P-type ATPase (ATP7B). Twelve different mutations have previously been identified in Taiwan Chinese with Wilson disease. We, herein, report another 4 missense mutations, 1 of which is novel. We did haplotype analysis of Taiwanese WND chromosomes, using three well characterized short tandem repeat markers (haplotype was assigned in the order of D13S314-D13S301-D13S316). Association correlation was found between the mutations and their respective haplotypes. Haplotype-deduced pedigree analysis was shown to be helpful in the mutation analysis of WND chromosomes and in the molecular assessment of both pre symptomatic WND patients and carriers. Given the complexity and heterogeneity of the mutation spectrum of ATP7B, we suggest that haplotype analysis should be performed before full-scale mutation analysis. PMID- 11043509 TI - Association of the vitamin D receptor start codon polymorphism (FokI) with bone mineral density in postmenopausal Korean women. AB - We undertook this study in order to examine the association between bone mineral density (BMD) and a polymorphism at the first of two potential translation initiation codons in the vitamin D receptor (VDR) gene. This polymorphism was detected by restriction fragment length polymorphism analysis, using polymerase chain reaction (PCR) and the restriction endonuclease FokI. The f allele indicates the presence of the FokI site, and the F allele its absence. The FokI genotype was determined in 174 postmenopausal Korean women, aged 43-71 years. The distribution of FokI genotypes in Koreans was found not to differ significantly from those found in Caucasians and Japanese, although it does differ significantly from that found in the black American population. We observed a significant association between the FokI polymorphism and lumbar BMD; P = 0.048, analysis of covariance [ANCOVA], but no association with femoral neck BMD (P = 0.505, ANCOVA). Those with the ff genotype had a 13.3% lower BMD in the lumbar spine than the FF subjects. In addition, a significantly higher prevalence of the ff genotype was observed in osteoporotic compared with osteopenic or normal women (P = 0.036, chi2 test). These data suggest that the ff genotype of the VDR gene correlates with decreased BMD in the lumbar spine in postmenopausal Korean women. PMID- 11043510 TI - Genomic structure of the gene encoding human 3-hydroxy-3-methyl-glutaryl coenzyme A reductase: comparison of exon/intron organization of sterol-sensing domains among four related genes. AB - We determined the genomic structure of the human gene encoding 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase, which catalyzes the conversion of HMG-CoA to mevalonate and is the rate-limiting and major regulatory enzyme in sterol biosynthesis. The gene is more than 21 kb long, about five times the size of its corresponding cDNA. It consists of 20 exons, ranging in size from 68 to 1809bp. An amino-terminal hydrophobic membrane-bound domain is encoded by exons 2 10, a flexible linker domain by exons 10 and 11, and the catalytic domain by exons 11-20. Exons 3-7 encode a sterol-sensing domain. We compared its genomic structure in this region with the sterol-sensing domains of three related genes, sterol regulatory element binding protein (SREBP) cleavage-activating protein (SCAP), Niemann-Pick type C1 protein (NPC1), and a morphogen receptor, Patched. Two of the five positions of introns in the sterol-sensing domain of the HMG-CoA reductase gene were identical to the exon/intron organization of this domain in the related human genes, but these positions of introns were not conserved in homologues from lower organisms, except in one instance. The data suggested that exon-shuffling may have occurred during relatively recent evolution: this would account for the structural similarity of this domain in four quite different human proteins. PMID- 11043511 TI - The human ribosomal protein L6 gene in a critical region for Noonan syndrome. AB - We have determined the genomic structure of the human ribosomal protein L6 gene (RPL6) and assigned it to the interval containing the Noonan syndrome locus. RPL6 spans 4415bp and consists of seven exons and six introns. The first exon is only 19bp in length, containing a 5' non-coding region and a polypyrimidine tract. The second exon starts with the initiator ATG. Although the overall structure of the protein is highly conserved among mammalian species, there is significant variation in the N-terminal portion. We have refined the position of RPL6, using two different radiation hybrid panels. RPL6 was mapped to chromosome 12q24.1 between the markers D12S84 and D12S861, which is in the critical region for Noonan syndrome. PMID- 11043512 TI - Association of essential hypertension in elderly Japanese with I/D polymorphism of the angiotensin-converting enzyme (ACE) gene. AB - Recent evidence suggests that an insertion/deletion (I/D) polymorphism of the gene encoding angiotensin-converting enzyme (ACE) is associated with myocardial infarction and related cardiovascular diseases. We investigated a possible association of the ACE polymorphism with essential hypertension in a total of 263 cases/controls from among the elderly (age, over 70 years) and middle-aged (age between 30 and 60 years) Japanese population. The frequency of the I/I homozygote was significantly higher in hypertensive subjects than in controls in the elderly age group (33/57 vs 16/46; P = 0.02), but no association was observed in the middle-aged group (25/75 vs 26/85; P = 0.71). Similarly, having at least one insertion allele was associated with essential hypertension in the elderly age group (83/114 vs 46/92 in controls; P = 0.001), but not in the middle-aged group (78/150 vs 94/170; P = 0.524). These data suggest that genetic variation at the ACE locus may be associated with some determinants for blood pressure in elderly persons, and imply the involvement of the ACE insertion/deletion polymorphism in the etiology of age-related essential hypertension in the Japanese population. PMID- 11043513 TI - Genomic structure and multiple single-nucleotide polymorphisms (SNPs) of the thiopurine S-methyltransferase (TPMT) gene. AB - Thiopurine S-methyltransferase (TPMT) catalyzes the S-methylation of drugs such as azathiopurine, 6-mercaptopurine, and 6-thioguanine, which are widely prescribed for immunosuppressive or cytotoxic applications. We report here the entire genomic structure of the TPMT gene and the presence of 30 single nucleotide polymorphisms (SNPs) within that structure. The gene spans a genomic region about 27kb long and consists of nine exons. By screening its entire genomic sequence for SNPs in 48 Japanese chromosomes by direct DNA sequencing, we detected 1 SNP in the 870-bp promoter region, 26 SNPs in introns, and 3 SNPs in the 3' untranslated region (3'UTR) for investigating correlations between TPMT genotypes and the side-effects caused by thiopurine drugs. PMID- 11043514 TI - Multiplex PCR amplification of TH01, D9S304, and D3S1744 loci. AB - A multiplex typing method of the tetrameric short tandem repeat (STR) loci TH11, D9S304, and D3S1744 was developed. The allelic ladder included alleles 6-11 (80 100bp) and 9.3 (95bp) for TH01, alleles 6-15 (125-161 bp) for D9S304, and alleles 13-22 (174-210bp) for D3S1744. The observed heterozygosity of D9S304 was 0.851. The combined discrimination power of the three loci was 0.991. PMID- 11043516 TI - cDNA cloning of a human RAB26-related gene encoding a Ras-like GTP-binding protein on chromosome 16p13.3 region. AB - Members of the RAB protein family are important regulators of vesicular fusion and trafficking. A putative new member of the RAB family of genes was identified through a public database search, and its full-length cDNA was isolated from a human fetal brain cDNA library. The predicted protein product of the gene consists of 190 amino acid residues and has 87% identity with rat Rab26. Thus, we designated this gene as the human RAB26-related gene. Reverse transcription coupled polymerase chain reaction (RT-PCR) demonstrated that the RAB26-related messenger RNA was predominantly expressed in adult and fetal brain. Furthermore, an RT-PCR experiment for brain subregions showed that the mRNA was highly expressed in the amygdala, cerebellum, caudate nucleus, and hippocampus. By PCR based analysis with both a human/rodent monochromosomal hybrid cell panel and a radiation hybrid panel, the gene was mapped to the chromosome 16p13.3 region between markers WI-7742 and WI-3061. The RAB26-related gene consists of eight exons that span about 44kb of the genome DNA. PMID- 11043515 TI - Genomic structure and chromosomal localization of the gene encoding TRAX, a Translin-associated factor X. AB - The TRAX gene encodes a Translin-associated 33-kDa protein partner, TRAX. The TRAX protein has extensive amino acid homology with Translin, and contains bipartite nuclear targeting sequences, suggesting a possible role in the selective nuclear transport of Translin lacking any nuclear targeting motifs. In the present study, genomic clones of the human TRAX gene were isolated to determine the complete genomic organization. The genomic structure of the human TRAX gene was similar to that of the human Translin gene, consisting of six exons and five introns, encompassing approximately 27kb in genomic DNA. Northern blot analysis revealed a predominant transcript of approximately 2.7kb, and its distribution in various tissues was like that of Translin. Chromosomal mapping by fluorescence in situ hybridization (FISH) analysis allowed localization of the TRAX gene to human chromosome lq41. PMID- 11043517 TI - Novel mutation of L718X in the ATP7A gene in a Japanese patient with classical Menkes disease, and four novel polymorphisms in the Japanese population. AB - Menkes disease is an X-linked recessive disorder of the copper membrane transport system caused by mutations in the ATP7A gene. While various mutations in the ATP7A gene have been reported, a genotype-phenotype correlation has not been clearly defined. A novel mutation in the ATP7A gene in a Japanese patient with classical Menkes disease was identified via analysis of reverse-transcriptase polymerase chain reaction products and genomic DNA of the ATP7A gene. The nonsense mutation, L718X, was found to result in premature termination and immature ATP7A protein, unlikely to have normal functioning. Therefore, this nonsense mutation of the ATP7A gene is proposed to play a causative role in presenting the classical Menkes phenotype. Furthermore, four novel polymorphisms, C1535T (L464L), C2151T (T669I), G2253A (R703H), and C3677T (H1178Y) were also identified. PMID- 11043518 TI - cDNA cloning of a new member of the Ras superfamily, RAB9-like, on the human chromosome Xq22.1-q22.3 region. AB - Members of the RAB protein family regulate vesicular trafficking and reside in specific intercellular compartments. A new member of the RAB family was identified through a public database search, and its full-length cDNA was isolated from a human fetal brain cDNA library. The predicted protein product of the gene consists of 201 amino acid residues, and the protein has 86% similarity to human RAB9 at the amino acid level. We designated the new gene RAB9-like. Northern blot analysis showed that the gene was transcribed ubiquitously in various human tissues. A database search revealed that the gene is divided into three exons and spans approximately 7.2kb of the genome DNA of chromosome Xq22.1 q22.3 region. PMID- 11043519 TI - Comparative studies for development of Schistosoma mansoni sporocysts in Puerto Rican and Brazilian Biomphalaria glabrata. AB - The development of sporocysts of Schistosoma mansoni was monitored in pigmented and albino Biomphalaria glabrata from Puerto Rico and Brazil. The snails were exposed individually to 20 miracidia, and sporocysts were allowed to develop for 3 to 12 weeks. Most of the immature sporocysts were found in the seminal receptacle sac and vas deferens during development. In contrast, mature daughter sporocysts were detected everywhere except in the foot at 12 weeks after exposure to the miracidia. It was found that mature daughter sporocysts formed more rapidly in the pigmented than in the albino snails, but no difference was observed in the formative time between the same types of Puerto Rican and Brazilian snails. It seems likely that there is a correlation between the infection rate and the time required for formation of mature daughter sporocysts in B. glabrata. PMID- 11043520 TI - Breath-hold gadolinium-enhanced three-dimensional MR thoracic aortography: higher spatial resolution imaging with phased-array coil and three-dimensional surface display. AB - The aim of this study was to examine signal intensities of data sets from MR thoracic aortography and to evaluate three-dimensional surface display (3DSD) for postprocessing. Twenty-five patients were imaged with gadolinium-enhanced 3D fast gradient echo sequence. The intensity at the aortic arch was significantly higher than that at the mediastinal fat (p<0.0001). The signal-to-noise ratio was lower at the aortic arch than at the ascending and descending aorta, whereas the contrast-to-noise ratio was fairly high at the aortic arch. Although in one case (4%) the intensity at the arch was smaller than that at the mediastinal fat, 3DSD was successfully performed in all cases. Superiority of 3DSD over maximum intensity projection was obtained in 67% of the cases. 3DSD was evaluated to be superior to maximum intensity projection in all cases of thoracic aortic aneurysm and coarctation of aorta. Higher resolution MR thoracic aortography could be successfully performed with phased-array coil and 3DSD. PMID- 11043521 TI - Results of filtering surgery in young patients with aniridia. AB - To evaluate whether filtering surgery is effective in controlling the intraocular pressure of young aniridic patients with glaucoma, we retrospectively reviewed the charts of aniridic patients with glaucoma under the age of 40 years. We defined a good intraocular pressure control period as the time from surgery until IOP exceeded 20 mm Hg, with or without glaucoma medication. Twenty filtering surgeries (17 trabeculectomies and 3 trabeculectomies with mitomycin C) were performed on 10 eyes in 6 patients for more than 20 years. The mean good intraocular pressure control period after the filtering surgery was 14.6 months (range, 2 to 54 months). Aside from mild choroidal detachment, no other serious complications were encountered. We believe that filtering surgery is efficacious for control of intraocular pressure of young aniridic patients with glaucoma. PMID- 11043522 TI - Ampullectomy of carcinoma of the papilla of vater in an elderly patient without jaundice. AB - A 79 year-old woman was admitted to Aioi City Hospital for a closer examination of hepatic dysfunction. A filling defect was observed at the distal end of the intrapancreatic common bile duct by computed tomography combined with drip infusion cholangiography. The diagnosis of adenoma with dysplasia at the papilla of Vater was obtained by a biopsy performed during duodenoscopy. As a result, we performed an ampullectomy. Histologic examination revealed a papillary adenocarcinoma which partly extended just beyond the muscle of Oddi. The patient made an uneventful recovery and was discharged on the 35th postoperative day. Here, based upon our experience, we discuss such problems as the accuracy of preoperative diagnosis and the indications for ampullectomy. PMID- 11043523 TI - Argentina at a crossroads in science. PMID- 11043524 TI - Serotonergic modulation of hippocampal acetylcholine release after long-term neuronal grafting. AB - Adult female rats sustained aspirative fimbria-fornix lesions and, 2 weeks later, received intrahippocampal grafts of fetal septal or mixed septal-raphe cell suspensions. Twenty-four months later, the extracellular concentration of hippocampal acetylcholine (ACh) was determined by microdialysis. Basal ACh levels (5-65 fmol/5 microl sham-operated rats) were strongly reduced after lesioning (3 7 fmol/5 microl). In septally transplanted and septal-raphe co-transplanted rats, hippocampal ACh concentrations were restored to near-normal levels (15-25 fmol/5 microl), indicating long-term functional survival of hippocampal transplants. After administration of citalopram (100 microM by infusion) and fenfluramine (20 mg/kg i.p.), the hippocampal ACh efflux was increased by 2- to 3-fold in all groups of rats. The relative increase of ACh was highest in co-transplanted rats, an effect which was possibly due to functional interactions between grafted raphe and septal neurons. PMID- 11043525 TI - AIDA reduces glutamate release and attenuates mechanical allodynia after spinal cord injury. AB - Spinal cord injury (SCI) leads to an increase in extracellular excitatory amino acid (EAA) concentrations, resulting in glutamate receptor-mediated excitotoxicity and central sensitization. To test contributions of group I metabotropic glutamate receptors (mGluRs) in SCI induced release of glutamate and in behavioral outcomes of central sensitization following injury, we administered 1-aminoindan-1,5-dicarboxylic acid (AIDA; 0.1 nmol intraspinally), a potent group I mGluR antagonist, to rats immediately after spinal cord contusion injury. EAAs were collected by microdialysis and quantified using HPLC. AIDA significantly decreased extracellular glutamate but not aspartate concentrations and significantly attenuated the development of mechanical but not thermal allodynia. These results suggest mGluRs play an important role in injury-induced EAA release and in central sensitization following SCI. PMID- 11043526 TI - Systemic nitroglycerin increases nNOS levels in rat trigeminal nucleus caudalis. AB - Systemic administration of nitroglycerin, a nitric oxide donor, triggers in migraineurs a delayed attack of unknown mechanisms. Subcutaneous nitroglycerin (10 mg/kg) produced a significant increase of nitric oxide synthase (NOS)- and c fos-immunoreactive neurons in the cervical part of trigeminal nucleus caudalis in rats after 4 h. This effect was not observed in the thoracic dorsal horn. Similar increase of NOS and c-fos was obtained in the brain stem after a somatic nociceptive stimulus, i.e. on the side of the formalin injection in the lip. Nitric oxide is thus able to increase NOS availability in second order nociceptive trigeminal neurons, which may be relevant for central sensitization and the understanding of its effect in migraine. PMID- 11043528 TI - Sensorimotor organization in patients who have undergone hemispherectomy: a study with (15)O-water PET and somatosensory evoked potentials. AB - To identify cortical structures that subserve residual motor and sensory function in patients with congenital hemiparesis due to a porencephalic cyst, we examined, using [(15)O]H2O, PET and somatosensory evoked potentials (SEPs) in three patients with left-sided hemiparesis who had undergone hemispherectomy. Motor stimulation of the affected hand produced ipsilateral activation in the premotor area in all patients, the SMA in two patients, and SII in two patients. Vibrotactile stimulation resulted in activation of the ipsilateral SII in all subjects. Median nerve stimulation of the affected hand produced ipsilateral long latency SEPs in fronto-centro-parietal areas, whereas stimulation of the non affected hand produced normal early cortical potentials in the contralateral hemisphere. Our results suggest that residual function in the paretic hand is warranted through non-primary motor and sensory areas, and higher order associative areas in the intact hemisphere. PMID- 11043527 TI - Enhanced motor cortical excitability in the unaffected hemisphere after hemispherectomy. AB - We evaluated motor cortical excitability of the unaffected hemisphere in three patients with intractable epilepsy who underwent hemispherectomy, using transcranial magnetic stimulation (TMS) and PET. TMS of the unaffected hemisphere evoked motor responses not only in the contralateral muscles but also in the ipsilateral ones in all the patients. A PET study in one patient showed activation of the unaffected motor cortex by movement of either arm. All of these responses were enhanced after the hemispherectomy, probably due to motor cortical disinhibition by transection of the corpus callosum. The PET study also showed postoperative activation of the premotor area of the unaffected hemisphere. These phenomena indicate posthemispherectomy neuroplastic reorganization leading to preservation of the motor function after the operation. PMID- 11043529 TI - Presenilin I interaction with cytoskeleton and association with actin filaments. AB - Presenilin I (PSI) has been shown to interact with microfilament-associated proteins of the filamin family. Here, we investigated a possible association of PSI with the cytoskeleton. Immunoblotting of detergent-insoluble fractions of rat brain homogenate revealed enrichment of neuron-specific 36 and 14 kDa proteolytic fragments of PSI, whereas 30 and 20 kDa fragments were found in the detergent soluble fraction. Specific severing of microfilaments with gelsolin in the detergent-insoluble pellet and subsequent centrifugation led to the detection of both actin and PSI fragments in the supernatant. In addition, in vitro translated PSI cosedimented with actin filaments. Our findings provide biochemical evidence for the association of PSI fragments with actin filaments. PMID- 11043530 TI - Immunohistochemical study of the hnRNP A2 and B1 in the rat forebrain. AB - Immunohistochemical techniques were employed to examine the distribution of RNA binding proteins A2 and B1 in the rat forebrain. Intense A2 and B1 immunolabeling were observed in the nucleoplasm of the neurons in the cerebral cortices, hippocampal formation, olfactory regions, caudate-putamen as well as the supraoptic nucleus of hypothalamus. In contrast, within the bed nucleus of the stria terminalis, as well as the medial and lateral habenular nucleus of thalamus, immunoreactivity for both proteins was weak. Within the globus pallidus and thalamic nucleus immunoreactivity for A2 was hardly detectable despite of intense B1 immunolabeling, while within the endopiriform nucleus and lateral and basolateral nucleus of amygdala intensity of B1 immunolabeling was relatively weak compared to A2. Our study suggests that the distribution of A2 and B1 are not constant throughout the forebrain and this diversity may reflect the post transcriptional regulation of cell-specific gene expression of neuronal cells. PMID- 11043531 TI - Inhibition of phosphatidylcholine synthesis precedes apoptosis induced by C2 ceramide: protection by exogenous phosphatidylcholine. AB - Cerebellar granule neurons in primary culture underwent apoptosis when exposed to C2-ceramide. Addition of exogenous phosphatidylcholine (PtdCho) resulted in a dose-dependent full prevention of neuronal death. Exogenous PtdCho also prevented apoptosis induced by farnesol, N-oleoylethanolamine, and sphingomyelinase, but did not prevent apoptosis induced after lowering the potassium concentration in the medium to non-depolarizing levels. Moreover, C2-ceramide inhibited labeling of [32P]PtdCho in cells incubated with [32P]orthophosphate, with the same potency to that causing apoptosis. Although cell viability did not decrease during the first few hours, inhibition of PtdCho synthesis was already patent after a 1 h exposure to C2-ceramide. Taken together, these results strongly suggest that inhibition of PtdCho synthesis constitutes one of the primary events by which C2 ceramide triggers apoptosis in cerebellar granule neurons. PMID- 11043532 TI - Maturation of frontal and temporal components of mismatch negativity (MMN) in children. AB - The mismatch negativity (MMN) response of auditory ERPs in adults appears to result from several overlapping components involving both frontal and temporal brain areas. Our aim was to test whether a similar configuration could be observed in children, and to examine the maturation rates of the different components. MMN (standard tones: 1000 Hz, deviants: 1100 Hz) was recorded from 28 scalp electrodes in 24 healthy children aged from 5 to 10 and in eight adults for comparison. Scalp current density analysis revealed both temporal and frontal components in children of all ages as well as in adults. Moreover the amplitudes of the temporal components were significantly greater in children than in adults, whereas the frontal components were similar at all ages. The results strongly suggest that MMN is mediated by at least two separate neural systems, and that the frontal system matures earlier than the sensory-specific system. PMID- 11043533 TI - Chronic morphine treatment inhibits oxytocin synthesis in rats. AB - The changes of oxytocin content and mRNA expression in some nuclei were investigated in morphine-dependent rats using radioimmunoassay (RIA) and in situ hybridization (ISH). After chronic administration of morphine, the oxytocin content in supraoptic nucleus (SON) and nucleus accumbens (NAc) decreased, and increased in the ventral tegment area (VTA) and locus coeruleus (LC), but did not change in other nuclei including the paraventricular nucleus (PVN), lateral septum (SEPTUM), raphe magnus nucleus (NRM) and periaquaductal gray (PAG). In morphine-L dependent rats, naloxone increased the levels of oxytocin in SON and PVN, but decreased that in LC. ISH first showed that chronic morphine treatment inhibited the oxytocin synthesis in SON but not in PVN. The present study demonstrates that chronic morphine treatment alters the brain oxytocin system, suggesting that oxytocin might contribute to the behavioral and neuroendocrine responses to morphine. PMID- 11043534 TI - Delivery of antisense oligonucleotides to neuroblastoma cells. AB - pH-Sensitive liposomes composed of dioleoylphosphatidylethanolamine and cholesterol hemisuccinate (3:2 mol/mol) were applied in delivery of antisense oligodeoxynucleotides (asODN) into NG 108-15 neuroblastoma and glioma cells. Fluorescently labelled asODN were entrapped in liposomes by a modified freeze thawing method (20% encapsulation efficiency). The uptake of asODN (free or entrapped in liposomes) by NG 108-15 cells was monitored by fluorescence activated cell sorting and confocal microscopy. Delivery of asODN was significantly improved when antisense were entrapped in liposomes as compared to free (nonliposomal) asODN. The uptake was dose-dependent and optimum was achieved after 2 h incubation. PMID- 11043535 TI - Sublethal hypoxia up-regulates corticotropin releasing factor receptor type 1 in fetal hippocampal neurons. AB - We analysed the influence of oxygen-glucose deprivation (OGD) on the expression of corticotropin releasing factor (CRF) receptors (CRF-R1 and CRF-R2) in fetal hippocampal neurons in vitro. A 2 h exposure of neurons to OGD resulted in death of 18+/-2.8% cells at 24 h following exposure, which was considered sublethal hypoxia. Expression of both receptors was quantitated by competitive reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Levels of mRNA for CRF-R1 were increased 3.2-fold compared to control neurons, while CRF-R2 mRNA levels remained unchanged. The increase of CRF-R1 mRNA levels was observed at 6 h and peaked at 24 h. CRF-R1 protein levels were also increased by 2.4-fold and 1.7-fold at 24 h and 48 h, respectively. These data suggest that the effects of CRF on neuronal survival are mediated in part through the induction and expression of CRF-R1 following a hypoxic/ischemic insult. PMID- 11043536 TI - The distribution of the neurokinin B receptor in the human and rat hypothalamus. AB - The neurokinin B receptor (NK3) is an element of the hypothalamic neuronal circuitry regulating blood pressure in rats. The present study used immunohistochemistry to reveal the distribution of NK3 in the human hypothalamus. The strongest NK3-like immunoreactivity in the human hypothalamus was found in neurons of the paraventricular nucleus, specifically in the parvicellular and posterior paraventricular subnuclei. Another prominent population of NK3-positive cells in the human hypothalamus was found in the perifornical nucleus. The present study also showed two previously unreported populations of NK3-positive neurons in the rat periventricular nucleus and medial magnocellular paraventricular subnucleus. It is concluded that there is a large degree of similarity in the distribution of NK3 in the human and rat hypothalamus. PMID- 11043538 TI - Neural cell adhesion molecules, CaM kinase II and long-term memory in the chick. AB - The intermediate and medial hyperstriatum ventrale (IMHV) of the chick brain is a site of recognition memory for filial imprinting. Previous results have demonstrated learning-related changes in the amounts of the three major isoforms of neural cell adhesion molecule (NCAM) in the left IMHV. The increases were present 24 h after training. The present study enquired whether the increases persisted and were present 48 h after training. The brain regions analysed were the left and right IMHV and the left and right hyperstriatum accessorium (HA), a visual projection area. The alpha-subunit of calcium/calmodulin protein kinase II (CaMKIIalpha) was also assayed. There were significant correlations between a measure of the strength of learning and the amount of NCAM 180 in the right IMHV (r = +0.65; p = 0.012) but not in the left, and in the left HA (r = -0.61; p = 0.02), but not in the right. There were no learning-related changes for CaMKIIalpha. We conclude that in IMHV the effects of imprinting on NCAM 180 are expressed mainly in the left IMHV 24 h after training, but 48 h after training are expressed mainly in the right IMHV. PMID- 11043537 TI - Gene expression of PSD95 in prefrontal cortex and hippocampus in schizophrenia. AB - A number of studies have suggested that disturbance in glutamatergic transmission in the cerebral cortex may underlie, or contribute to the pathophysiology of schizophrenia. In this study we examined expression of the postsynaptic density protein 95 (PSD95) mRNA in the prefrontal cortex and hippocampus in postmortem material from neuroleptic-treated schizophrenics and normal controls. PSD95 is known to bind to NMDA receptor subunits and is known to be involved in synaptic plasticity. In situ hybridization analysis showed that the expression of PSD95 was significantly decreased in Brodmann area 9 of the prefrontal cortex but not in the hippocampus. These results further implicate the prefrontal cortex in the pathophysiology of schizophrenia and suggest dysfunction of NMDA receptors in the schizophrenic cortex. PMID- 11043539 TI - Configural face processes in acquired and developmental prosopagnosia: evidence for two separate face systems? AB - Configural face processes were tested using face recognition and face detection tasks in a comparison of acquired and developmental prosopagnosia. In the recognition task the two patients showed a very different pattern. The developmental patient does not show an inversion effect while the acquired prosopagnosia patient is better at matching inverted than normal stimuli. Moreover, there is no effect of face context on matching features in the developmental case while the acquired prosopagnosia patient shows a strong negative effect of context. However, in a speeded face detection task both patients are similarly unimpaired. The results are consistent with the existence of two separate face systems, one involved in face detection and the other in face recognition. PMID- 11043540 TI - Reactive astrocytes show enhanced inwardly rectifying K+ currents in situ. AB - Injury and diseases of the nervous system can induce astrocytes to form tenacious glial scars. We induced focal cortical freeze-lesions in neonatal rats and examined scars histologically and electrophysiologically in tissue slices isolated 2-3 weeks after lesioning. Lesions displayed marked gliosis, characterized by upregulation of GFAP labeling. Reactive astrocytes surrounding the scar showed marked hypertrophy, enlarged cell bodies and extended processes frequently terminating with endfeet-like structures on blood vessels. These reactive astrocytes showed enhanced expression of inwardly rectifying K+ (K(IR)) channels, widely believed to be an important pathway for astrocytic K+ buffering. These results suggest that a subpopulation of reactive astrocytes along a glial scar might be instrumental in buffering K+ away from the lesion. PMID- 11043541 TI - 12-O-tetradecanoyl-phorbol-13-acetate down-regulates the Huntingtin promoter at Sp1 sites. AB - We have studied the effects of the phorbol ester, 12-O-tetradecanoyl-phorbol-13 acetate (TPA) on Huntington's disease (HD) gene transcription in neuronal and non neuronal cell lines, to investigate pathways regulating HD gene expression. TPA reduced transcription from the HD gene promoter in SK-N-SH (neuroblastoma) and HeLa cells but not in JEG3 (choriocarcinoma) cells. In SK-N-SH cells, the responsible cis-acting promoter sequences comprise the tandemly duplicated Sp1 sites in the region from -213 to -174, relative to the translation start site. The TPA-down-regulating region in HeLa cells was mapped to the sequence from -141 to -126. In conclusion, this demonstrates that HD gene transcription can be down regulated in vitro in a cell-specific manner. PMID- 11043542 TI - Rapid reduction in somatostatin mRNA expression by hypothalamic neurons induced by dexamethasone. AB - We have previously reported that peripherally administered dexamethasone induces a rapid increase in hypothalamic somatostatin release. Here we investigated whether somatostatin synthesis could also be affected by this treatment and the potential involvement of glutamate in this effect. Male rats received a saline or a dexamethasone injection (300 microg/100 g body weight) and were killed 30 min later. Thirty minutes prior to dexamethasone treatment, another group received an i.p. injection of MK-801, a NMDA receptor antagonist. Cells expressing somatostatin mRNA in the periventricular nucleus were analyzed by in situ hybridization using digoxigenin-labeled somatostatin oligonucleotide probe. Dexamethasone decreased the number of digoxigenin-labeled cells expressing somatostatin mRNA in the periventricular nucleus as compared to the same histological sections from control rats. The dexamethasone effect was reversed by pretreatment with MK-801, which alone also decreased the number of cells expressing somatostatin mRNA. In summary, dexamethasone administration induces a significant rapid decrease in periventricular cells expressing somatostatin mRNA and this effect is partly abolished by MK-801. PMID- 11043543 TI - Subunit composition of rat ventral spinal cord GABA(A) receptors, assessed by single cell RT-multiplex PCR. AB - We analyzed the expression of native GABA(A) receptors in choline acetyltransferase and glutamic acid decarboxilase positive cells, from lamina IX of the lumbar region of rat spinal cord. More than one isoform of each subunit was detected within a single cell. The alpha3, alpha5, alpha1, beta3 and gamma2 subunit was the most frequent combination in both cell populations. However, the total number of subunit expressed by each cell type was different, being the ChAT positive cells the simplest. Interestingly, the ChAT and GAD positive cells also displayed a different pattern of distribution of both spliced isoforms of the gamma2 subunit. These results indicate that several GABA(A) receptors, with different molecular composition, are expressed in a single cell and that different cell types can express different GABA(A) receptors. PMID- 11043544 TI - Functional sex differences in the accessory olfactory bulb of the rat. AB - The aim of this study is to determine whether sex-related differences exist in the biosynthetic activity of the mitral cells within the mitral layer of the AOB. Possible functional changes over the estrus cycle and the potential effects of castration and androgenization are assessed. Biosynthetic activity was measured using silver staining of the argyrophilic proteins associated with the nucleolar organizer regions (Ag-NOR). Assisted by stereological methods, the following parameters were studied: mean number, percentage and mean area of Ag-NOR in estrus and diestrus females, intact males, castrated and androgenizated rats. We detected sex differences in a histochemical marker related to synthetic activity, an estrus cycle effect and changes resulting from the perinatal treatments. We conclude that this structurally dimorphic region is also functionally dimorphic. PMID- 11043546 TI - ERP analysis of cognitive sequencing: a left anterior negativity related to structural transformation processing. AB - A major objective of cognitive neuroscience is to identify those neurocomputational processes that may be shared by multiple cognitive functions vs those that are highly specific. This problem of identifying general vs specialized functions is of particular interest in the domain of language processing. Within this domain, event related brain potential (ERP) studies have demonstrated a left anterior negativity (LAN) in a range 300-700 ms, associated with syntactic processing, often linked to grammatical function words. These words have little or no semantic content, but rather play a role in encoding syntactic structure required for parsing. In the current study we test the hypothesis that the LAN reflects the operation of a more general sequence processing capability in which special symbols encode structural information that, when combined with past elements in the sequence, allows the prediction of successor elements. We recorded ERPs during a non-linguistic sequencing task that required subjects (n = 10) to process special symbols possessing the functional property defined above. When compared to ERPs in a control condition, function symbol processing elicits a left anterior negative shift between temporal and spatial characteristics quite similar to the LAN described during function word processing in language, supporting our hypothesis. These results are discussed in the context of related studies of syntactic and cognitive sequence processing. PMID- 11043545 TI - Mouse Tspan-5, a member of the tetraspanin superfamily, is highly expressed in brain cortical structures. AB - Using a subtractive hybridization method for the identification of genes related to the development of the murine cerebral cortex, we cloned a mouse homologue of a human tetraspanin family member, Tspan-5. We have isolated a 3.1 Kb cDNA fragment containing the entire coding region. Analysis of the cDNA nucleotide sequence revealed that mouse Tspan-5 shares 98% amino acid sequence identity with its human homologue. The predicted length of the mouse protein is 268 amino acids, with four putative hydrophobic domains with N- and C-intracellular tails, and two extracellular domains. Northern blot analysis of adult mouse tissues showed a single transcript, which is preferentially expressed in the brain. In situ hybridization showed prominent expression of Tspan-5 in the neocortex, the hippocampus, amygdala and in Purkinje cells in the cerebellum. The pattern of expression of Tspan-5 in the mouse brain suggests a role for the tetraspanins in the maintenance of adult brain function. PMID- 11043547 TI - Repetitive peripheral magnetic stimulation alleviates tactile extinction. AB - Despite its frequency in right brain damaged patients crucial mechanisms of tactile extinction are still obscure and treatments are unavailable. Recent PET observations suggest a hypometabolism in the primary and secondary somatosensory cortex of the lesioned hemisphere in patients with tactile extinction. Functional and morphological investigations have shown that the sensorimotor cortex has a remarkable capability of reorganization when the sensory inflow is changed. Repetitive peripheral magnetic stimulation (RPMS) applied in patients suffering from central paresis alleviates sensorimotor as well as cognitive deficits by the induction of proprioceptive inflow, thereby activating plasticity in the CNS. Based on the observation of reduced metabolic activity in patients suffering from tactile extinction we applied RPMS to explore the effects of peripheral sensory stimulation on tactile extinction. Fourteen right-hemisphere lesioned patients with tactile extinction were randomly allocated to an experimental and a control group. The experimental group received one single RPMS treatment of the left forearm as well as a condition of attentional cueing known to improve visual extinction. The control group, with comparable tactile extinction scores, neither received RPMS nor verbal cueing, but was tested twice to evaluate possible learning or test repetition effects. In the experimental group RPMS led to a significant reduction of left-sided extinctions in the recognition of different tactual surfaces, but had no effect on ipsilesional errors. In contrast, attentional cueing had no significant effect on left-sided extinction errors but unexpectedly increased right-hand extinction errors slightly but significantly. The control group showed stable extinction scores of the left- and right-hand stimulus across two measurements, thus ruling out learning or test repetition effects. These results show that sensory inflow is an important modulatory factor in tactile extinction. Furthermore, multiple RPMS may prove a promising way for the rehabilitation of patients with this disorder. PMID- 11043548 TI - Role of Ca2+-permeable non-NMDA glutamate receptors in spinal nociceptive transmission. AB - The functional role of Ca2+-permeable non-NMDA receptors in spinal nociceptive processing was investigated using joro spider toxin (JSTx), a selective blocker of these receptors. JSTx 0.25 and 1 microg administered spinally produced a significant facilitation of the C-fibre evoked response and post-discharge, but not the A-fibre response, of dorsal horn neurones recorded in adult rats. This may result from a block of Ca2+-permeable AMPA receptors located on GABAergic interneurones. At higher doses, this facilitation of responses was lost, suggesting additional Ca2+-permeable non-NMDA receptors, possibly kainate receptors, in excitatory spinal pathways. Thus, functional Ca2+-permeable AMPA receptors are present within the dorsal horn, predominantly within inhibitory pathways, and play a role distinct from other excitatory amino acid receptors in spinal nociceptive processing. PMID- 11043549 TI - Agmatine improves locomotor function and reduces tissue damage following spinal cord injury. AB - Clinically effective drug treatments for spinal cord injury (SCI) remain unavailable. Agmatine, an NMDA receptor antagonist and inhibitor of nitric oxide synthase (NOS), is an endogenous neuromodulator found in the brain and spinal cord. Evidence is presented that agmatine significantly improves locomotor function and reduces tissue damage following traumatic SCI in rats. The results suggest the importance of future therapeutic strategies encompassing the use of single drugs with multiple targets for the treatment of acute SCI. The therapeutic targets of agmatine (NMDA receptor and NOS) have been shown to be critically linked to the pathophysiological sequelae of CNS injury and this, combined with the non-toxic profile, lends support to agmatine being considered as a potential candidate for future clinical applications. PMID- 11043550 TI - Kindled seizure-evoked somatostatin release in the hippocampus: inhibition by MK 801. AB - The aim of this study was to evaluate the contribution of ionotropic glutamate receptors to kindled seizure-evoked somatostatin release in the hippocampus, using a microdialysis approach. Basal and amygdala stimulation-evoked somatostatin-like immunoreactivity (-LI) release was significantly greater in kindled compared to naive rats. In naive rats, neither hippocampal perfusion with the selective AMPA/kainate receptor antagonist GYKI 52466 nor with the selective NMDA receptor antagonist MK-801 affected behavior, EEG, or somatostatin-LI release. In kindled rats, GYKI 52466 was still devoid of any effect, while MK-801 significantly decreased stimulus-evoked (but not basal) somatostatin-LI efflux. MK-801 produced identical effects when injected i.p. This study provides the first direct evidence that kindled seizure-evoked somatostatin release in the hippocampus is partly NMDA receptor dependent. PMID- 11043551 TI - Deciphering the role of Gi2 in opioid-induced adenylyl cyclase supersensitization. AB - Prolonged opioid treatment of HEK 293 cells expressing opioid receptors are known to induce adenylyl cyclase supersensitization, a process that requires pertussis toxin (PTX)-sensitive G(i/o) proteins. Here, the role of Gi2 in adenylyl cyclase supersensitization was investigated. A PTX-insensitive G alpha(i2)/z chimera was stably co-expressed with mu-, kappa- or delta-opioid receptors in HEK 293 cells. Functional coupling of G alpha(i2)/z to the opioid receptors was demonstrated by opioid-induced inhibition of adenylyl cyclase and stimulation of ERK1/2 phosphorylation in PTX-treated cells. Chronic opioid treatment of each cell line led to adenylyl cyclase supersensitization but this response was blocked by PTX. Our results demonstrated that although PTX-sensitive G proteins are obligatory for opioid-induced adenylyl cyclase supersensitization, Gi2 alone was insufficient to mediate this response. PMID- 11043552 TI - Recurrent mossy fibers preferentially innervate parvalbumin-immunoreactive interneurons in the granule cell layer of the rat dentate gyrus. AB - Detection of vesicular zinc and immunohistochemistry against markers for different interneuron subsets were combined to study the postsynaptic target selection of zinc-containing recurrent mossy fiber collaterals in the dentate gyrus. Mossy fiber collaterals in the granule cell layer selectively innervated parvalbumin-containing cells, with numerous contacts per cell, whereas the granule cells were avoided. Under the electron microscope, those boutons made asymmetrical contacts on dendrites and somata. These findings suggest that, in addition to the hilar perforant path-associated (HIPP) interneurons, the basket and chandelier cells also receive a powerful feed-back drive from the granule cells, and thereby are able to control population synchrony in the dentate gyrus. On the other hand, the amount of monosynaptic excitatory feed-back among granule cells is shown to be negligible. PMID- 11043553 TI - Mutation of the conserved N-terminal cysteine (Cys92) of human presenilin 1 causes increased A beta42 secretion in mammalian cells but impaired Notch/lin-12 signalling in C. elegans. AB - The presenilin proteins are involved in the proteolytic processing of transmembrane proteins such as Notch/lin-12 and the beta-amyloid precursor protein (betaAPP). Mutation of a conserved cysteine (Cys60Ser) in the C. elegans presenilin sel-12 has a loss-of-function effect on Notch/lin-12 processing similar to that of null mutations in sel-12. In contrast, in mammalian cells, most missense mutations increase gamma-secretase cleavage of betaAPP. We report here that mutation of this conserved cysteine (Cys92Ser) in human presenilin 1 confers a loss-of-function effect in C. elegans, but causes increased A beta42 secretion in mammalian cells. These data suggest that the role of presenilins in Notch/lin-12 signalling and betaAPP processing are either separately regulated activities or independent activities of the presenilins. PMID- 11043554 TI - The calcium receptor modulates the hyperpolarization-activated current in subfornical organ neurons. AB - Here we report that neurons of the subfornical organ (SFO), a circumventricular structure devoid of a blood-brain barrier, show time-dependent, inward rectification indicative of the presence of a subthreshold, hyperpolarization activated inward current (Ih). In whole-cell patch clamp experiments of isolated SFO neurons, we observed a Cs+-sensitive Ih in 47% of cells tested. Furthermore, we show that Ih is involved in the generation of evoked bursts in SFO neurons. An allosteric agonist of the extracellular calcium-sensing receptor (CaR) was found to potentiate Ih consistent with our previous observations of CaR-mediated bursting in SFO neurons. These studies indicate that a proportion of SFO neurons express Ih, and this may be one ionic mechanism through which bursting is regulated by various extracellular messengers. PMID- 11043555 TI - Serotonergic dorsal raphe neurons cease firing by disfacilitation during paradoxical sleep. AB - Using in vivo extracellular unit recordings combined with microdialysis infusion in the cat, we found that the cessation of discharge of presumed serotonergic dorsal raphe neurons during paradoxical sleep (PS) was completely blocked by either histamine or phenylephrine, an alpha1 adrenoceptor agonist, but not by bicuculline, a GABA receptor antagonist. In addition, application of mepyramine, a specific H1 histamine receptor antagonist, or prazosin, a specific alpha1 adrenoceptor antagonist, suppressed the spontaneous discharge of raphe neurons during both quiet waking and sleep. The present data suggest that this cessation of dorsal raphe unit activity is caused by the mechanism of disfacilitation resulting from the cessation of discharge of norepinephrine- or histamine containing neurons during PS. PMID- 11043556 TI - Regulation of nociceptin mRNA expression in the septum by dopamine and adenosine systems. AB - Most effects of nociceptin are related to blockade of stress and anxiolytic-like effects. This neuropeptide is highly expressed in septal nuclei, which are involved in response to stressful situations. Dopamine and adenosine may have modulatory effects on stress behaviour by acting on septal neurons. We therefore analysed the regulation of septal nociceptin expression using quantitative in situ hybridization following manipulations of adenosine and dopamine neurotransmission. No difference was observed between wild-type and A2A receptor deficient mice. In both genotypes, chronic treatments with caffeine, an equipotent A1 and A2A adenosine receptor antagonist, did not significantly modify nociceptin expression. 6-Hydroxydopamine-induced dopamine depletion was also without effect. These results demonstrate that dopamine and adenosine are not involved in the regulation of septal nociceptin expression in spite of the involvement of these three neurotransmitters in stress and anxiety behaviours. PMID- 11043557 TI - Direct mapping of ocular dominance columns in human primary visual cortex. AB - Functional magnetic resonance imaging at 2.0T was employed to identify columnar structures in human visual cortex. Sagittal sections (4 mm thickness) covering the calcarine cortex were acquired with use of a multiecho low flip angle gradient-echo sequence at 4.0 s temporal resolution and 0.25 x 0.25 mm2 spatial resolution. Extending earlier attempts based on a differential paradigm contrasting left vs right eye stimulation, this work presents the first direct mapping of human ocular dominance columns by measuring separate activation maps with left and right eye stimulation. The resulting individual maps reveal patterns of ocular dominance as spots or bands of altered activity in calcarine cortex. Their superposition shows only little spatial overlap of eye-specific encoding which strongly supports the genuineness of these functional units. PMID- 11043559 TI - Current orientation induced by magnetic stimulation influences a cognitive task. AB - The direction of the current induced by transcranial magnetic stimulation (TMS) over the motor cortex has been observed to influence the threshold and latency of evoked muscle responses. This study investigates the effect of TMS-induced current orientation (ICO) over the prefrontal cortex, on a specific cognitive task (memory-guided saccade). TMS was applied with a figure-of-eight coil, placed at one of eight different orientations over the prefrontal cortex. The most effective ICO was antero-lateral, which is a different optimal ICO from that seen over the hand area of the motor cortex. This demonstrates that ICO can alter the effect of TMS on cognitive functions and that ICO is an independent variable that should not be ignored when designing TMS studies. PMID- 11043558 TI - Effect of CART in the hypothalamic paraventricular nucleus on feeding and uncoupling protein gene expression. AB - Cocaine and amphetamine regulated transcript (CART) decreases feeding and body weight after ventricular injection. CART mRNA and peptide are found in the paraventricular nucleus of the hypothalamus (PVN). The purpose of the present study was to determine effects of PVN-injected CART on feeding and thermogenic capacity. PVN-injected CART (55-102, 100 pmol) significantly decreased NPY induced feeding at 1, 2 and 4 h, but did not significantly affect deprivation induced feeding. CART induced gene expression of uncoupling protein 1 (UCP1), UCP2, and UCP3 in brown and white adipose tissue and biceps femoris muscle respectively. These results indicate the PVN as a specific site of CART action, and suggest that CART in the PVN may have an important influence on energy metabolism. PMID- 11043560 TI - Lithium lengthens the circadian period of individual suprachiasmatic nucleus neurons. AB - Lithium treatment lengthens the period of circadian rhythms in most organisms. In the present study, we tested whether lithium acts directly on the mammalian suprachiasmatic nucleus (SCN) to lengthen rhythms of individual neurons. Lithium increased the circadian period of firing rate rhythms of cultured SCN neurons in a concentration-dependent manner. Lithium had no effect on the amplitude of these rhythms, but did affect the period of some cells more than others. The results indicate that lithium acts directly on the SCN to lengthen the free-running period of individual neurons. PMID- 11043561 TI - Differential somato-dendritic localization of TrkA, TrkB, TrkC and p75 mRNAs in vivo. AB - TrkB mRNA was shown to be localized in the somatodendritic compartment in vitro but no data are currently available on the subcellular distribution of the neurotrophin receptors mRNAs in vivo. Here we describe the subcellular distribution of TrkA, TrkB, TrkC and p75 mRNAs in the adult rat basal forebrain. We find that TrkA, TrkC and p75 mRNAs are restricted to the cell soma but in addition, p75 mRNA labelling extends in average for 8 microm within the proximal dendrites of 34% of the labelled neurons. TrkB mRNA has a somatodendritic distribution in 95% of the labelled neurons reaching variable distances in different neurons (23-84.5 microm) and forebrain regions (mean: 40, 51 and 55 microm for diagonal band, septum and ventral pallidum). PMID- 11043562 TI - Magnetically induced phosphenes in sighted, blind and blindsighted observers. AB - Direct stimulation of visual cortex can produce illusory flashes of light, called phosphenes. Here we describe the spatial and motion properties of phosphenes produced by transcranial magnetic stimulation in normal subjects and in two subjects with peripheral or cortical blindness. The totally retinally blind subject experienced normal phosphenes, apart from their concentration in the centre of the visual field, whereas the hemianopic subject, lacking area V1, did not experience phosphenes when his surviving extrastriate visual areas were stimulated. In the absence of V1, magnetically induced activity was unable to generate a conscious visual percept in the field defect. PMID- 11043563 TI - Ipsilateral and contralateral subthalamic activity after unilateral dopaminergic lesion. AB - Unilateral lesions of the dopaminergic nigral neurons in rats are currently used as a model of Parkinson's disease. However, several neurochemical studies have questioned the possible influence of the lesioned side on the contralateral non lesioned side. To address this question, electrophysiological recordings in the ipsilateral and contralateral subthalamic nucleus was performed on anaesthetized rats, 3, 7 and 14 days after induction of a unilateral dopaminergic lesion. At these three times, the mean discharge rate of the subthalamic neurons recorded ipsilateral to the lesion was increased by 85, 176 and 127%, respectively, whereas this rate was decreased by 16, 27 and 43%, respectively, in the opposite subthalamic nucleus. This result emphasizes the importance of interhemispheric regulation of this structure, contrasting with the unilateralized current model of the functional organization of the basal ganglia. PMID- 11043564 TI - Ischemic tolerance affects the adenylation state of GluR2 mRNA. AB - To analyse GluR2 regulations in the rat hippocampal CA1 region following global and tolerance-inducing ischemia in situ hybridization (ISH) and quantitative PCR (Q-PCR) was applied. In addition, cDNA was synthesised from two different primer combinations in order to elucidate possible differences in the adenylation state of GluR2 mRNA. Following global ischemia, ISH and Q-PCR both showed reductions to half of control levels of GluR2 mRNA in consent with previously published results. Following tolerance induction, ISH showed no changes, whereas PCR analysis showed up-regulation to 228% of control value for the general cDNA synthesis, and no change for the specific cDNA synthesis. This indicates that tolerance-inducing ischemia does not increase the amount of GluR2 mRNA; instead polyadenylation of the existing GluR2 mRNA pool takes place. PMID- 11043565 TI - Theta rhythm increases in left superior temporal cortex during auditory hallucinations in schizophrenia: a case report. AB - Auditory hallucinations (AH), the perception of sounds and voices in the absence of external stimuli, remain a serious problem for a large subgroup of patients with schizophrenia. Functional imaging of brain activity associated with AH is difficult, since the target event is involuntary and its timing cannot be predicted. Prior efforts to image the patterns of cortical activity during AH have yielded conflicting results. In this study, MEG was used to directly image the brain electrophysiological events associated with AH in schizophrenia. We observed an increase in theta rhythm, as sporadic bursts, in the left superior temporal area during the AH states, whereas there was steady theta band activity in the resting state. The present finding suggests strong association of the left superior temporal cortex with the experience of AH in this patient. This is consistent with the hypothesis that AH arises from areas of auditory cortex subserving receptive language processing. PMID- 11043566 TI - Differential expression of mRNAs of GDNF family in the striatum following 6-OHDA induced lesion. AB - Changes of mRNA levels of GDNF and its recently discovered congeners persephin, neurturin, artemin in the striatum of lesioned side following 6-hydroxydopamine (6-OHDA) lesion in rodents were investigated with semi-quantitative RT-PCR. Two weeks after the lesions were made, mRNA levels of GDNF family members, except for neurturin, were significantly increased in the striatum on the side ipsilateral to the lesion compared with equivalent tissue of sham control. This increase reached a maximal level 5-7 weeks post-lesion. The marked increase of BDNF mRNA expression was also observed in the ipsilateral striatum with similar time course. These findings suggest that 6-OHDA-induced lesions can change gene expression in denervated target tissue, and that mRNA levels of GDNF family members in striatal cells may be modulated by afferent dopaminergic input in a slow-rising and long-lasting fashion. PMID- 11043567 TI - Detection of determinism within time series of irregular burst firing from the injured sensory neuron. AB - Irregular burst firing is spontaneously generated by rat dorsal root ganglion neuron after chronic compression. To investigate the time series of burst firing, we recorded interspike intervals (ISIs) of single-unit firing in vitro and processed the ISIs to obtain interevent intervals (IEIs). Then, two non-linear methods were applied to detect deterministic dynamic behaviors in IEI series. No evidence for the existence of determinism was found with non-linear prediction method. Using unstable periodic orbit identification method, significant period-1 orbits were identified in all 10 data, period-2 orbits in eight, and period-3 orbits in six. The results indicate that there exist significant deterministic behaviors in the time series of irregular burst firing from the injured sensory neuron. PMID- 11043568 TI - Cerebral localization of the center for reading and writing music. AB - The mechanisms that underlie the ability to read and write music remain largely unclear compared to those involved in reading and writing language. We had the extremely rare opportunity to study the cerebral localization of the center for reading and writing music in the case of a professional trombonist. During rehearsal immediately before a concert, he suffered a hemorrhage that was localized to the left angular gyrus, the area that has long been known as the center for the ability to read and write. Detailed tests revealed that he showed symptoms of alexia with agraphia for both musical scores and language. PMID- 11043569 TI - Primary structure and function of the catecholamine release inhibitory peptide catestatin (chromogranin A(344-364)): identification of amino acid residues crucial for activity. AB - The novel chromogranin A fragment catestatin (bovine chromogranin A(344-364); RSMRLSFRARGYGFRGPGLQL) is a potent inhibitor of catecholamine release (IC50, approximately 0.2-0.3 microM) by acting as a nicotinic cholinergic antagonist. To define the minimal active region within catestatin, we tested the potencies of synthetic serial three-residue deletion (amino-terminal, carboxyl-terminal, or bidirectional) fragments to inhibit nicotine-stimulated catecholamine secretion from PC12 pheochromocytoma cells. The results revealed that a completely active core sequence of catestatin was constituted by chromogranin A(344-364). Nicotinic cationic signal transduction was affected by catestatin fragments in a manner similar to that for secretion (confirming the functional importance of the amino terminus). To identify crucial residues within the active core, we tested serial single amino acid truncations or single residue substitutions by alanine on nicotine-induced catecholamine secretion and desensitization. Nicotinic inhibition by the active catestatin core was diminished by even single amino acid deletions. Selective alanine substitution mutagenesis of the active core revealed important roles for Met346, Leu348, Phe350, Arg351, Arg353, Gly354, Tyr355, Phe357, and Arg358 on catecholamine secretion, whereas crucial roles to inhibit desensitization of catecholamine release were noted for Arg344, Met346, Leu348, Ser349, Phe350, Arg353, Gly354, Tyr355, Gly356, and Arg358. We conclude that a small, 15-amino acid core of catestatin (chromogranin A(344-364)) is sufficient to exert the peptide's typical inhibitory effects on nicotinic cholinergic stimulated catecholamine secretion, signal transduction, and desensitization. These studies refine the biologically active domains of catestatin and suggest that the pharmacophores for inhibition of nicotinic secretion and desensitization may not be identical. PMID- 11043570 TI - The antiangiogenic factor 16K PRL induces programmed cell death in endothelial cells by caspase activation. AB - We asked whether the antiangiogenic action of 16K human PRL (hPRL), in addition to blocking mitogen-induced vascular endothelial cell proliferation, involved activation of programmed cell death. Treatment with recombinant 16K hPRL increased DNA fragmentation in cultured bovine brain capillary endothelial (BBE) and human umbilical vein endothelial (HUVE) cells in a time- and dose-dependent fashion, independent of the serum concentration. The activation of apoptosis by 16K hPRL was specific for endothelial cells, and the activity of the peptide could be inhibited by heat denaturation, trypsin digestion, and immunoneutralization, but not by treatment with the endotoxin blocker, polymyxin B. 16K hPRL-induced apoptosis was correlated with the rapid activation of caspases 1 and 3 and was blocked by pharmacological inhibition of caspase activity. Caspase activation was followed by inactivation of two caspase substrates, poly(ADP-ribose) polymerase (PARP) and the inhibitor of caspase activated deoxyribonuclease (DNase) (ICAD). Furthermore, 16K hPRL increased the conversion of Bcl-X to its proapoptotic form, suggesting that the Bcl-2 protein family may also be involved in 16K hPRL-induced apoptosis. These findings support the hypothesis that the antiangiogenic action of 16K hPRL includes the activation of programmed cell death of vascular endothelial cells. PMID- 11043571 TI - A synthetic triterpenoid, 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO), is a ligand for the peroxisome proliferator-activated receptor gamma. AB - A novel synthetic triterpenoid, 2-cyano-3,12-dioxooleana-1,9-dien-28-oic acid (CDDO), previously reported to have potent differentiating, antiproliferative, and antiinflammatory activities, has been identified as a ligand for the peroxisome proliferator-activated receptor gamma (PPARgamma). CDDO induces adipocytic differentiation in 3T3-L1 cells, although it is not as potent as the full agonist of PPARgamma, rosiglitazone. Binding studies of CDDO to PPARgamma using a scintillation proximity assay give a Ki between 10(-8) to 10(-7) M. In transactivation assays, CDDO is a partial agonist for PPARgamma. The methyl ester of CDDO, CDDO-Me, binds to PPARgamma with similar affinity, but is an antagonist. Like other PPARgamma ligands, CDDO synergizes with a retinoid X receptor (RXR) specific ligand to induce 3T3-L1 differentiation, while CDDO-Me is an antagonist in this assay. The partial agonism of CDDO and the antagonism of CDDO-Me reflect the differences in their capacity to recruit or displace cofactors of transcriptional regulation; CDDO and rosiglitazone both release the nuclear receptor corepressor, NCoR, from PPARgamma, while CDDO-Me does not. The differences between CDDO and rosiglitazone as either partial or full agonists, respectively, are seen in the weaker ability of CDDO to recruit the coactivator CREB-binding protein, CBP, to PPARgamma. Our results establish the triterpenoid CDDO as a member of a new class of PPARgamma ligands. PMID- 11043573 TI - Differential regulation of mitogen-activated protein kinase-responsive genes by the duration of a calcium signal. AB - We have investigated the cellular mechanisms by which changes in intracellular calcium (Ca2+) can differentially regulate gene expression. Two Ca2+ paradigms, involving prolonged and transient Ca2+ increases, were used. As a starting point, we studied the slow, prolonged elevation of Ca2+ caused by activation of 5-HT1 receptors. We had previously shown that 5-HT1 agonists inhibit calcitonin gene related peptide (CGRP) transcription and secretion. The Ca2+ ionophore, ionomycin, was used to produce a prolonged elevation of the Ca2+ signal similar to that generated by 5-HT1 receptor agonists. Ionomycin treatment of the neuronal like CA77 cell line specifically inhibited mitogen-activated protein (MAP) kinase stimulation of the CGRP enhancer and two synthetic MAP kinase-responsive reporter genes (4- to 10-fold). We then showed that ionomycin repression of promoter activity involved selective induction of MAP kinase phosphatase-1 (MKP-1), but not MKP-2, and that overexpression of MKP-1 was sufficient to repress CGRP enhancer activity. These effects were then compared with a Ca2+ paradigm involving a transient elevation in Ca2+ as seen after depolarization. At 4 h after the transient increase in Ca2+, the CGRP enhancer and synthetic MAP kinase responsive reporter genes were stimulated. In contrast, exposure to depolarizing stimuli overnight caused only a less than 2-fold inhibition of promoter activity. We propose that the duration of the Ca2+ signal can determine the magnitude of a negative feedback loop that leads to differential regulation of MAP kinase responsive genes. PMID- 11043572 TI - Tumor necrosis factor alpha-mediated insulin resistance, but not dedifferentiation, is abrogated by MEK1/2 inhibitors in 3T3-L1 adipocytes. AB - Tumor necrosis factor-alpha (TNFalpha) has been implicated as a contributing mediator of insulin resistance observed in pathophysiological conditions such as obesity, cancer-induced cachexia, and bacterial infections. Previous studies have demonstrated that TNFalpha confers insulin resistance by promoting phosphorylation of serine residues on insulin receptor substrate 1 (IRS-1), thereby diminishing subsequent insulin-induced tyrosine phosphorylation of IRS-1. However, little is known about which signaling molecules are involved in this process in adipocytes and about the temporal sequence of events that ultimately leads to TNFalpha-stimulated IRS-1 serine phosphorylation. In this study, we demonstrate that specific inhibitors of the MAP kinase kinase (MEK)1/2-p42/44 mitogen-activated protein (MAP) kinase pathway restore insulin signaling to normal levels despite the presence of TNFalpha. Additional experiments show that MEK1/2 activity is required for TNFalpha-induced IRS-1 serine phosphorylation, thereby suggesting a mechanism by which these inhibitors restore insulin signaling. We observe that TNFalpha requires 2.5-4 h to markedly reduce insulin triggered tyrosine phosphorylation of IRS-1 in 3T3-L1 adipocytes. Although TNFalpha activates p42/44 MAP kinase, maximal stimulation is observed within 10 30 min. To our surprise, p42/44 activity returns to basal levels well before IRS 1 serine phosphorylation and insulin resistance are observed. These activation kinetics suggest a mechanism of p42/44 action more complicated than a direct phosphorylation of IRS-1 triggered by the early spike of TNFalpha-induced p42/44 activity. Chronic TNFalpha treatment (>> 72 h) causes adipocyte dedifferentiation, as evidenced by the loss of triglycerides and down-regulation of adipocyte-specific markers. We observe that this longer term TNFalpha-mediated dedifferentiation effect utilizes alternative, p42/44 MAP kinase-independent intracellular pathways. This study suggests that TNFalpha-mediated insulin resistance, but not adipocyte dedifferentiation, is mediated by the MEK1/2-p42/44 MAP kinase pathway. PMID- 11043574 TI - Identification and characterization of constitutively active Smad2 mutants: evaluation of formation of Smad complex and subcellular distribution. AB - Smads mediate activin, transforming growth factor beta (TGFbeta), and bone morphogenetic protein signaling from receptors to nuclei. According to the current model, activated activin/TGFbeta receptors phosphorylate the carboxyl terminal serines of Smad2 and Smad3 (SSMS-COOH); phosphorylated Smad2/3 oligomerizes with Smad4, translocates to the nucleus, and modulates transcription of defined genes. To test key features of this model in detail, we explored the construction of constitutively active Smad2 mutants. To mimic phosphorylated Smad2, we made two Smad2 mutants with acidic amino acid substitutions of carboxyl terminal serines: Smad2-2E (Ser465, 467Glu) and Smad2-3E (Ser464, 465, 467Glu). The mutants enhanced basal transcriptional activity in a mink lung epithelial cell line, L17. In a Smad4-deficient cell line, SW480.7, Smad2-2E did not affect basal signaling; however, cotransfection with full-length Smad4, but not transfection of Smad4 alone, resulted in enhanced basal transcriptional activity, suggesting that the constitutively active Smad2 mutant also requires Smad4 for function. In vitro protein interaction analysis revealed that Smad2-2E bound more tightly to Smad4 than did wild-type Smad2; dissociation constants were 270 +/- 66 nM for wild-type Smad2:Smad4 complexes and 79 +/- 18 nM for Smad2-2E:Smad4 complexes. Determination of the subcellular localization of Smad2 revealed that a greater percentage of Smad2-2E was localized in the nucleus than wild-type Smad2. These results suggest that Smad2 phosphorylation results in both tighter binding to Smad4 and increased nuclear concentration; those changes may be responsible for transcriptional activation by Smad2. PMID- 11043575 TI - Prolactin stimulates activation of c-jun N-terminal kinase (JNK). AB - In recent years the mitogen-activated protein (MAP) kinase family has expanded to include both c-jun N-terminal kinases (JNKs), and the p38/HOG1 family in addition to the extracellular regulated kinase (ERK) family. These kinases are activated by a variety of growth factors, as well as extra- and intracellular insults such as osmotic stress, UV light, and chemotherapeutic agents. Stimulation of the PRL dependent Nb2 cell line with PRL results in the rapid activation of JNK as determined by the glutathione-S-transferase (GST)-jun kinase assay. Activation was maximal 30 min after stimulation with 50 nM rat PRL (rPRL) and decreased after that time. Dose response studies indicated that concentrations as low as 10 nM rPRL resulted in maximal activation. The interleukin-3 (IL-3)-dependent myeloid progenitor cell line 32Dcl3 was transfected with the long, Nb2, and short forms of the rat PRL receptor (rPRLR), as well as the long form of the human PRLR (hPRLR). The long and Nb2 forms of the PRLR were able to stimulate activation of JNK; however, the short form of the rPRLR was not. This corresponds with the inability of the short form of the rPRLR to stimulate proliferation of 32Dcl3 cells. Activation of JNK in 32Dcl3 cells expressing the long form of the hPRLR was maximal at 30 min after stimulation with 100 nM ovine PRL (oPRL) and declined after that time. Dose response studies indicated that activation of JNK was maximal after 30 min at a concentration of 10 nM, and the amount of activated JNK declined at the highest concentration of oPRL, 100 nM. Immunoblot analysis with an antibody that recognizes the activated (phosphorylated) forms of JNK1 and JNK2 indicated that both JNK1 and JNK2 isoforms were activated in 32D/hPRLR cells stimulated with oPRL. A recombinant human adenovirus expressing a kinase-inactive mutant of JNK1 (APF mutant) was used to determine the biological effect of blocking JNK activity in Nb2 cells. Expression of the JNK1-APF mutant inhibited cellular proliferation and induced DNA fragmentation typical of cells undergoing apoptosis. These data suggest that activation of JNKs may be important in mitogenic signaling and/or suppression of apoptosis in Nb2 cells. PMID- 11043576 TI - Mutational analysis of the androgen receptor AF-2 (activation function 2) core domain reveals functional and mechanistic differences of conserved residues compared with other nuclear receptors. AB - A short C-terminal sequence that forms the core of the activation function-2 (AF 2) domain is conserved in members of the nuclear receptor superfamily and is required for their normal biological function. Despite a high degree of sequence similarity, there are differences in the context and structure of AF-2 in different nuclear receptors. To gain deeper insight into these differences, we carried out an extensive mutational analysis of the AF-2 core in the androgen receptor (AR) and compared the changes in transcriptional activity with similar mutations that have previously been generated in other nuclear receptors. Mutagenesis of Met894 to Asp resulted in substantial decreases in both DNA and ligand binding activities and, consequently, a significant drop in ligand dependent transcriptional activation. In contrast, substitution of Met894 with Ala did not affect DNA or hormone binding, and the transactivation potential was comparable to that of wild-type AR. Mutagenesis of Glu897 either with Val or Ala significantly impaired ligand-dependent activation that was not due to changes in DNA or ligand binding. There are both similarities and distinct differences between these findings compared with previous mutagenesis studies of the corresponding residues in other nuclear receptors. All mutants efficiently interfered with AP-1 activity, indicating that ligand-dependent activation of transcription and interference with AP-1 activity are separable functions in AR. For the Glu897 substitutions, the decrease in ligand-dependent transactivation could partially be reversed by overexpression of GRIP1 (GR-interacting protein 1) or CBP, putative coactivators for AR. However, there was no correlation between ligand-dependent in vitro or in vivo association with coactivators and the ability of the mutants to support ligand-dependent transactivation. This is in contrast to similar mutations in other nuclear receptors that lose interactions with putative coactivators concomitant with their loss of transcriptional activity. However, the Glu897 mutations disrupted the intramolecular interaction between the N-terminal domain and the ligand-binding domain of AR that was recently suggested to be required for normal AR function. We conclude that residues in the AF-2 core domain of AR make distinctly different contributions to its transcriptional activities compared with those of other nuclear receptors studied to date. PMID- 11043577 TI - Androgen receptor nuclear translocation is facilitated by the f-actin cross linking protein filamin. AB - The human androgen receptor (hAR) is a ligand-dependent transcription factor responsible for the development of the male phenotype. The mechanism whereby nuclear translocation of the hAR is induced by its natural ligand 5alpha dihydrotestosterone is a phenomenon not fully understood. The two-hybrid interaction trap assay has been used to isolate proteins that interact with the hAR in an attempt to identify molecules involved in hAR transactivation and movement. We have identified the actin-binding protein filamin, a 280-kDa component of the cytoskeleton, as an hAR interacting protein. This interaction is ligand independent but is enhanced in its presence. The functional significance of this interaction was analyzed using a cell line deficient in filamin via transient expression of a green fluorescent protein-hAR chimera. In filamin deficient cells this revealed that hAR remained cytoplasmic even after prolonged exposure to synthetic ligand. Nuclear shuttling was restored when this cell line regained wild-type expression of filamin. These data suggest a novel role for filamin, implicating it as an important molecule in AR movement from the cytoplasm to the nucleus. PMID- 11043578 TI - The LIM/homeodomain protein islet-1 modulates estrogen receptor functions. AB - LIM/Homeodomain (HD) proteins are classically considered as major transcriptional regulators which, in cooperation with other transcription factors, play critical roles in the developing nervous system. Among LIM/HD proteins, Islet-1 (ISL1) is the earliest known marker of motoneuron differentiation and has been extensively studied in this context. However, ISL1 expression is not restricted to developing motoneurons. In both embryonic and adult central nervous system of rodent and fish, ISL1 is found in discrete brain areas known to express the estrogen receptor (ER). These observations led us to postulate the possible involvement of ISL1 in the control of brain functions by steroid hormones. Dual immunohistochemistry for ISL1 and ER provided evidence for ISL1-ER coexpression by the same neuronal subpopulation within the rat hypothalamic arcuate nucleus. The relationship between ER and ISL1 was further analyzed at the molecular level and we could show that 1) ISL1 directly interacts in vivo and in vitro with the rat ER, as well as with various other nuclear receptors; 2) ISL1-ER interaction is mediated, at least in part, by the ligand binding domain of ER and is significantly strengthened by estradiol; 3) as a consequence, ISL1 prevents ER dimerization in solution, thus leading to a strong and specific inhibition of ER DNA binding activity; 4) ISL1, via its N-terminal LIM domains, specifically inhibits the ER-driven transcriptional activation in some promoter contexts, while ER can serve as a coactivator for ISL1 in other promoter contexts. Taken together, these data suggest that ISL1-ER cross-talk could differentially regulate the expression of ER and ISL1 target genes. PMID- 11043579 TI - Estrogen-induced activation of Erk-1 and Erk-2 requires the G protein-coupled receptor homolog, GPR30, and occurs via trans-activation of the epidermal growth factor receptor through release of HB-EGF. AB - Estrogen rapidly activates the mitogen-activated protein kinases, Erk-1 and Erk 2, via an as yet unknown mechanism. Here, evidence is provided that estrogen induced Erk-1/-2 activation occurs independently of known estrogen receptors, but requires the expression of the G protein-coupled receptor homolog, GPR30. We show that 17beta-estradiol activates Erk-1/-2 not only in MCF-7 cells, which express both estrogen receptor alpha (ER alpha) and ER beta, but also in SKBR3 breast cancer cells, which fail to express either receptor. Immunoblot analysis using GPR30 peptide antibodies showed that this estrogen response was associated with the presence of GPR30 protein in these cells. MDA-MB-231 breast cancer cells (ER alpha-, ER beta+) are GPR30 deficient and insensitive to Erk-1/-2 activation by 17beta-estradiol. Transfection of MDA-MB-231 cells with a GPR30 complementary DNA resulted in overexpression of GPR30 protein and conversion to an estrogen responsive phenotype. In addition, GPR30-dependent Erk-1/-2 activation was triggered by ER antagonists, including ICI 182,780, yet not by 17alpha-estradiol or progesterone. Consistent with acting through a G protein-coupled receptor, estradiol signaling to Erk-1/-2 occurred via a Gbetagamma-dependent, pertussis toxin-sensitive pathway that required Src-related tyrosine kinase activity and tyrosine phosphorylation of tyrosine 317 of the Shc adapter protein. Reinforcing this idea, estradiol signaling to Erk-1/-2 was dependent upon trans-activation of the epidermal growth factor (EGF) receptor via release of heparan-bound EGF (HB EGF). Estradiol signaling to Erk-1/-2 could be blocked by: 1) inhibiting EGF receptor tyrosine kinase activity, 2) neutralizing HB-EGF with antibodies, or 3) down-modulating HB-EGF from the cell surface with the diphtheria toxin mutant, CRM-197. Our data imply that ER-negative breast tumors that continue to express GPR30 may use estrogen to drive growth factor-dependent cellular responses. PMID- 11043580 TI - Hypoxia prevents induction of aromatase expression in human trophoblast cells in culture: potential inhibitory role of the hypoxia-inducible transcription factor Mash-2 (mammalian achaete-scute homologous protein-2). AB - The human placenta has a remarkable capacity to aromatize C19-steroids, produced by the fetal adrenals, to estrogens. This reaction is catalyzed by aromatase P450 (P450arom), encoded by the CYP19 gene. In placenta, CYP19 gene expression is restricted to the syncytiotrophoblast layer. Cytotrophoblasts isolated from human placenta, when placed in monolayer culture in 20% O2, spontaneously fuse to form syncytiotrophoblast. These morphological changes are associated with a marked induction of aromatase activity and CYP19 gene expression. When cytotrophoblasts are cultured in an atmosphere containing 2% O2, they manifest increased rates of DNA synthesis and fail to fuse and form syncytiotrophoblast. The objective of the present study was to utilize cytotrophoblasts isolated from midterm human placenta to analyze the effects of O2 on CYP19 gene expression and the molecular mechanisms that mediate these effects. We observed that when trophoblast cells were maintained in 2% O2, there was only a modest induction of CYP19 expression as a function of time in culture, and aromatase activity was barely detectable. However, when cytotrophoblasts that had been maintained in 2% O2 for 3 days were placed in a 20% O2 environment, there was a rapid onset of cell fusion and induction of P450arom mRNA and aromatase activity. In addition, mRNAs for the helix-loop-helix factors Mash-2 (mammalian achaete-scute homologous protein-2) and Id1 (inhibitor of differentiation 1) were readily detectable in freshly isolated cytotrophoblasts and were markedly decreased upon differentiation to syncytiotrophoblast in 20% O2. By contrast, when cytotrophoblasts were cultured in 2% O2, mRNA levels for Mash-2 and Id1 remained elevated. Interestingly, overexpression of Mash-2 in primary cultures of human trophoblast cells markedly inhibited cell fusion and the spontaneous induction of P450arom mRNA levels and caused a marked decrease in expression of co-transfected fusion gene constructs containing either 125, 201, 246, or 501 bp of DNA flanking the 5'-end of the placenta-specific exon (exon I.1) of the human CYP19 gene linked to the human GH (hGH) structural gene, as reporter. In studies using BeWo, a human choriocarcinoma cell line, overexpression of Mash-2 also inhibited expression of cotransfected CYP19I.1:hGH fusion gene constructs. The findings that Mash-2 had no effect on the expression of a CYP19I.1(-42):hGH fusion gene in primary cultures of human trophoblast and BeWo cells suggest that Mash-2 exerts its inhibitory effects directly or indirectly though CYP19I.1 5'-flanking sequences that lie between -42 and -125 bp. By contrast, neither Id1 nor Id2 had an effect on CYP19I. 1 promoter activity in the transfected BeWo cells. These findings suggest that Mash-2 may serve as a hypoxia-induced transcription factor that prevents differentiation to syncytiotrophoblast and aromatase induction in human trophoblast cultured under low O2 conditions. PMID- 11043581 TI - Conditional disruption of the aryl hydrocarbon receptor nuclear translocator (Arnt) gene leads to loss of target gene induction by the aryl hydrocarbon receptor and hypoxia-inducible factor 1alpha. AB - To determine the function of the aryl hydrocarbon receptor nuclear translocator (ARNT), a conditional gene knockout mouse was made using the Cre-loxP system. Exon 6, encoding the conserved basic-helix-loop-helix domain of the protein, was flanked by loxP sites and introduced into the Arnt gene by standard gene disruption techniques using embryonic stem cells. Mice homozygous for the floxed allele were viable and had no readily observable phenotype. The Mx1-Cre transgene, in which Cre is under control of the interferon-gamma promoter, was introduced into the Arnt-floxed mouse line. Treatment with polyinosinic polycytidylic acid to induce expression of Cre resulted in complete disruption of the Arnt gene and loss of ARNT messenger RNA (mRNA) expression in liver. To determine the role of ARNT in gene control in the intact animal mouse liver, expression of target genes under control of an ARNT dimerization partner, the aryl hydrocarbon receptor (AHR), was monitored. Induction of CYP1A1, CYP1A2, and UGT1*06 mRNAs by the AHR ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin was absent in livers of Arnt-floxed/Mx1-Cre mice treated with polyinosinic-polycytidylic. These data demonstrate that ARNT is required for AHR function in the intact animal. Partial deletion of the Arnt allele was found in kidney, heart, intestine, and lung. Despite more than 80% loss of the ARNT expression in lung, maximal induction of CYP1A1 was found, indicating that the expression level of ARNT is not limiting to AHR signaling. Cobalt chloride induction of the glucose transporter-1 and heme oxygenase-1 mRNAs was also markedly abrogated in mice lacking ARNT expression, suggesting an inhibition of HIF-1alpha activity. These studies establish a critical role for ARNT in AHR and HIF-1alpha signal transduction in the intact mouse. PMID- 11043582 TI - Identification of a short cis-acting element in the human vasopressin type 2 receptor gene which confers high-level expression of a reporter gene specifically in collecting duct cells. AB - In the kidney, water reabsorption is mainly regulated by the binding of arginine vasopressin to vasopressin type 2 (V2) receptors. These receptors are expressed selectively in principal cells of the collecting ducts. To identify molecular mechanisms responsible for the cell-specific expression of the V2 receptor, we have analyzed the proximal promoter of the corresponding gene. We report the identification of a 33-bp enhancer [collecting duct tissue-specific element 1 (CSE1)] that induced high levels of expression of the luciferase reporter gene in three collecting duct cell lines, but not in other renal cell lines. In gel shift assays, CSE1 bound a DNA-binding protein expressed selectively in collecting duct cell lines, and a 7-bp mutation, which abolished the activity of CSE1 in transient transfection experiments, also abolished the binding of this protein. Furthermore, decoy experiments performed using CSE1 showed that this sequence was involved not only in the expression of a construct containing 4.2 kb of the V2 receptor proximal promoter, but also in the expression of the endogenous V2 receptor gene. CSE1 appears to act mostly by counteracting the inhibitory effects of a strong ubiquitous repressor element that we called CIE1. Collectively, these results identify the first functional collecting duct-specific cis-acting element. PMID- 11043583 TI - Applications of mass spectrometry to food proteins and peptides. AB - The application of mass spectrometry (MS) to large biomolecules has been revolutionized in the past decade with the development of electrospray ionization (ESI) and matrix-assisted laser desorption/ionization (MALDI) techniques. ESI and MALDI permit solvent evaporation and sublimation of large biomolecules into the gaseous phase, respectively. The coupling of ESI or MALDI to an appropriate mass spectrometer has allowed the determination of accurate molecular mass and the detection of chemical modification at high sensitivity (picomole to femtomole). The interface of mass spectrometry hardware with computers and new extended mass spectrometric methods has resulted in the use of MS for protein sequencing, post translational modifications, protein conformations (native, denatured, folding intermediates), protein folding/unfolding, and protein-protein or protein-ligand interactions. In this review, applications of MS, particularly ESI-MS and MALDI time-of-flight MS, to food proteins and peptides are described. PMID- 11043584 TI - Preparation and evaluation of packed capillary columns for the separation of nucleic acids by ion-pair reversed-phase high-performance liquid chromatography. AB - Oligonucleotides and double stranded DNA fragments were separated in 200 microm I.D. capillary columns packed with micropellicular, octadecylated, 2.1 microm poly(styrene-divinylbenzene) particles by ion-pair reversed-phase high performance liquid chromatography (IP-RP-HPLC). Both the length and the diameter of the connecting capillaries (150 x 0.020 mm I.D.) as well as the detection volume (3 nl) had to be kept to a minimum in order to maintain the high efficiency of this chromatographic separation system with peak widths at half height in the range of a few seconds. Three different types of frits, namely sintered silica particles, sintered octadecylsilica particles, and monolithic poly(styrene-divinylbenzene) (PS-DVB) frits were evaluated with respect to their influence on chromatographic performance. Best performance for the separation of oligonucleotides and long DNA fragments was observed with the PS-DVB frits, whereas the short DNA fragments were optimally resolved in columns terminated by octadecylsilica frits. The maximum loading capacity of 60 x 0.20 mm I.D. columns ranged from 20 fmol (7.7 ng) for a 587 base pair DNA fragment to 500 fmol (2.4 ng) for a 16-mer oligonucleotide. Lower mass- and concentration detection limits in the low femtomol and low nanomol per liter range, respectively, make capillary IP-RP-HPLC with UV absorbance detection highly attractive for the separation and characterization of minute amounts of synthetic oligonucleotides, DNA restriction fragments, and short tandem repeat sequences amplified by polymerase chain reaction. PMID- 11043585 TI - Investigation of mandelic acid bonding on Pirkle type chromatographic stationary phases by Raman spectroscopy. AB - The bonding of mandelic acid enantiomers has been studied on benzene-leucine, dinitrobenzene-leucine and dinitrobenzene-phenylalanine type chiral stationary phases connected to zeolite A supports. The pi-donor, pi-acceptor and H-bonding interactions responsible for diastereomer pair formations can be studied under quasi in situ chromatographic conditions by Fourier transform Raman and surface enhanced Raman spectroscopic techniques. Structural differences between diastereomer pairs result in observable spectral differences at a phase load of approx. 50%. It was shown that the decreasing pi-acceptor character of the phase is associated with its increasing capability of H-bond formation. Correlating spectral data to chromatographic results it can be concluded that, in addition to H-bonding as well as to pi-donor-pi-acceptor interactions, steric hindrances due to bulky moieties of either the stationary phase or the analyte molecules are of importance in successful separations. PMID- 11043586 TI - Separation of the four pairs of enantiomers of vincamine alkaloids by enantioselective high-performance liquid chromatography. AB - The four enantiomeric pairs of vincamine group alkaloids were separated by HPLC using Chiralpak AD as chiral stationary phase (CSP) and various n-hexane-2 propanol and n-hexane-ethanol mobile phases. (+)-cis-Vincamine, which is used in pharmaceutical preparations, is eluted much faster than its optical isomer, with separation factors of 2.4 and 3.5, respectively in these mobile phases. Other CSPs gave negative results. A chiral recognition mechanism is proposed and circular dichroism spectra of the individual enantiomers are presented. PMID- 11043587 TI - Comparison of different liquid chromatography methods for the determination of corticosteroids in biological matrices. AB - Various extraction techniques can be combined with column liquid chromatography (LC) and ultraviolet (UV) or mass spectrometric (MS) detection for the determination of synthetic corticosteroids in biological matrices. Target analysis of low concentrations of 25 microg/kg of dexamethasone in feed can be performed by combining immunoaffinity chromatography (IAC) and LC with UV detection. A straightforward multi-analyte procedure is obtained by tandem solid phase extraction (SPE) and subsequent LC-UV. However, the limits of detection for feed samples are then relatively poor, viz. 100 microg/kg. A multi-analyte method which meets modern demands of about 5 microg/kg detection limit requires one-step SPE combined with LC-MS analysis. As regards urine corticosteroids can be determined down to a level of 0.5 microg/l by either SPE-LC-MS- MS or SPE(IAC)-LC MS. PMID- 11043588 TI - Derivatization of ephedrine with o-phthaldialdehyde for liquid chromatography after treatment with sodium hypochlorite. AB - The usefulness of the reaction with NaClO followed by derivatization with o phthaldialdehyde (OPA) and N-acetyl-L-cysteine (NAC) has been investigated for the chromatographic analysis of ephedrine. The influence of parameters affecting the two-stage reaction has been evaluated, including concentration of NaClO, time of reaction, temperature and pH. On the basis of these studies, conditions for the pre-column and (automated) post-column determination of ephedrine are presented. The described conditions have been applied to the measurement of ephedrine in the concentration intervals 0.2-20.0 microg/ml and 2.0-50.0 microg/ml for the pre-column and post-column methods, respectively. The possibility of applying the NaClO/ OPA-NAC method to other primary, secondary and tertiary derivative amphetamines has also been evaluated. PMID- 11043589 TI - Development of simultaneous purification methodology for multiple synthetic peptides by reversed-phase sample displacement chromatography. AB - We have developed a low-pressure protocol, designed as a rapid, simple and cost effective procedure for the efficient and parallel purification of multiple peptide mixtures. This was achieved through adaptation of our novel reversed phase sample displacement chromatography (SDC) method, where the major separation process takes place in the absence of organic modifier, to modular solid-phase extraction (SPE) technology. Thus, crude peptide sample is applied at overload conditions to extraction columns consisting of SPE tubes containing silica-based reversed-phase packing. By applying a vacuum to draw the solution through the packing, product separation from hydrophobic and hydrophilic impurities is accomplished in a two-stage purification unit: a short pre-column functions as a trap for hydrophobic impurities, while a second, longer SPE column is used as a product isolation column. Thus, under ideal SDC conditions, washing with a 100% aqueous solvent will achieve retention of hydrophobic impurities on the trap, with displacement of product and hydrophilic impurities from the trap to the product isolation column; hydrophilic impurities are thus displaced off the product isolation to waste, leaving only product retained on the main column. In this initial evaluation, this purification system has demonstrated excellent separation of product, in good yield, from both hydrophilic and hydrophobic impurities over a wide range of peptide hydrophobicity and crude composition for model synthetic peptide systems representing crude peptide mixtures. PMID- 11043590 TI - Analysis of a peptide hormone mixture of therapeutic interest by liquid chromatography coupled to high-flow pneumatically assisted electrospray mass spectrometry. AB - High-flow pneumatically assisted electrospray ionization mass spectrometry (ESI MS) has been extensively used for the characterization and determination of peptides and peptide hormones available for biomedical research and therapeutic applications. The aim of this study was to optimize a method of characterization and determination of a mixture of peptide hormones with therapeutic interest by liquid chromatography (LC) coupled to ESI-MS. In this work the linear solvation energy relationship methodology was used in order to optimize the mobile phase to be used in the LC separation of the peptide hormone series and the operational parameters of the source and analyzer of ESI were also optimized to obtain the best signal stability and the highest sensitivity. To validate the proposed method for peptide hormone analysis, quality parameters were determined and satisfactory results were obtained. Likewise, the method detection limit was picomole level for most of the peptides employing selected-ion monitoring of the [M+nH]n+ ions. PMID- 11043591 TI - Purification of substance P endopeptidase activity in the rat ventral tegemental area with the Akta-Purifier chromatographic system. AB - The new chromatographic system Akta-Purifier 10 (Amersham-Pharmacia Biotech), scaled for preparative HPLC, was used for the purification of Substance P (SP) endopeptidase activity in the ventral tegemental area (VTA) of the rat brain. SP endopeptidase previously identified and purified from human cerebrospinal fluid has been found to degrade the neuroactive peptide SP in a specific pattern. In this study we have recovered SP endopeptidase from the rat VTA following a purification scheme involving homogenization (ultrasonication) and extraction of the excised tissue, size-exclusion chromatography (Superdex 75 HR), and ion exchange chromatography (Resource Q). In this way we were able to achieve a purification factor of almost 7,500, based on specific activity. The obtained SP endopeptidase activity, was then subjected to characterization with regard to inhibition profile. The enzyme activity was monitored by following the conversion of SP to its N-terminal fragment SP(1-7) using a radioimmunoassay, specific for the heptapeptide product. On basis of inhibition profile it was possible to discern two different SP endopeptidase-like activities, one sensitive toward the protease inhibitor phosphoramidon (preparation A), and another non-sensitive to phosphoramidon or captopril (preparation B). The molecular masses of preparations A and B, as derived from sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were found to be 90,000 and 76,000, respectively. Our data suggest that the purified phosphoramidon sensitive endopeptidase activity may be an enzyme that plays a major role in the conversion of SP to its bioactive fragment SP(1-7) in the rat VTA. This is likely to be identical to the previously known neutral endopeptidase (EC 3.4.24.11). However, this study also demonstrates the existence of a distinct endopeptidase activity with properties in agreement with rat spinal cord SP endopeptidase. In the context of previously shown altered levels of SP(1-7) in the VTA during morphine withdrawal both purified enzyme activities may turn out to be responsible. PMID- 11043592 TI - Effects of DNA topology, temperature and solvent viscosity on DNA retardation in slalom chromatography. AB - Slalom chromatography is a unique size-fractionation method applicable to large DNA molecules [>5 kilobase pairs (kbp)]. The method was first developed by using columns packed with microbeads (diameter, <20 microm) used for high-performance liquid chromatography and by applying a relatively fast flow-rate (>0.3 ml/min). Previous studies suggested that the separation is attributed to a hydrodynamic rather than to an equilibrium phenomenon (J. Hirabayashi and K. Kasai, Anal. Biochem. 178 (1989) 336; J. Hirabayashi, N. Itoh, K. Noguchi and K. Kasai, Biochemistry, 29 (1990) 9515). In the present report, the results of a systematic study on the effects of DNA topology, temperature, and solvent viscosity on DNA retardation are described. Firstly, the behaviour of circular (super-coiled) and linearized forms of charomid DNAs (20-42 kbp) was studied. Circular-form DNA molecules were found to be fractionated size-dependently similarly to linear forms in a flow-rate dependent manner. However, the extent of retardation of the circular form DNA was apparently less than that of the corresponding linear forms. Circular DNAs showed almost the same retardation (e.g., 42 kbp) as DNA fragments (e.g., 20 kbp) having approximately half of the size of the former. This observation indicates that DNA retardation is basically related to physical length, not to mass. Secondly, to study the effect of temperature with special reference to solvent viscosity, we carried out chromatographic analysis at various temperatures ranging from 6 to 65 degrees C in both the absence and presence of sucrose (10 or 20%, w/v). The results showed that it is the solvent viscosity that determines the extent of retardation. Taken together, all of physicochemical parameters that define hydrodynamic properties, i.e., particle size, flow-rate and solvent viscosity, proved to be critical in slalom chromatography as well as the potential physical length of the DNA, thus supporting the concept that slalom chromatography is based on a hydrodynamic principle. PMID- 11043593 TI - Determination of thiophanate-methyl and its metabolites at trace level in spiked natural water using the supported liquid membrane extraction and the microporous membrane liquid-liquid extraction techniques combined on-line with high performance liquid chromatography. AB - On-line supported liquid membrane (SLM) extraction and microporous membrane liquid-liquid extraction (MMLLE) techniques for sample preparation of natural water samples have been developed for the determination of thiophanate-methyl (TM), carbendazim (MBC) and 2-aminobenzimidazole (2-AB) using reversed-phase HPLC. The combination of SLM extraction and MMLLE offers extraction conditions that makes it possible to determine a wide variety of compounds, i.e., permanently charged, ionisable and non-polar at sub ppb level. The detection limits obtained after extraction are about 0.1 microg/l for MBC and 2-AB using SLM, and 0.5 x Lg/l for TM using MMLLE and the precision is better than 5% for both systems. Typical enrichment rates are 0.6 and 2.7 times/min using SLM and MMLLE, respectively. PMID- 11043594 TI - Determination of ammonium in milk and dairy products by ion chromatography. AB - To control the quality and the biochemical evolution of milk and dairy products during their technological transformations, it is interesting to determine their ammonium concentrations. A chromatographic method for the determination of this compound is proposed. The method is based on the separation of ammonium by cation exchange chromatography and its detection by suppressed conductivity. With an appropriate sample preparation, this method enabled identification and quantification of ammonium with good repeatability (relative standard deviation of about 5%). Moreover, good sensitivity (less than 0.5 mg/kg) and no interference between ammonium and other matrix components were determined. It was also shown that this method offers a very promising alternative for studying changes in ammonium concentration of milk or caseinate after their heat treatments and in different dairy products such as yoghurt and cheeses (hard cooked and mould ripened cheeses). PMID- 11043595 TI - Determination of fungicides in natural waters using solid-phase microextraction and gas chromatography coupled with electron-capture and mass spectrometric detection. AB - This study develops a method for the analysis of seven fungicides in environmental waters, using solid-phase microextraction (SPME). The analyzed compounds--dicloran, chlorothalonil, vinclozolin, dichlofluanid, captan, folpet and captafol--belong to different classes of chemical compound (chloroanilines, sulphamides, phthalimides and oxazolidines) and are used mainly in agriculture and as antifouling paints. Their determination was carried out by gas chromatography with electron-capture and mass spectrometric detection. To perform SPME, four types of fibre have been assayed and compared: polyacrylate (85 microm), polydimethylsiloxane (100 and 30 microm), carbowax-divinylbenzene (CW DVB 65 microm) and polydimethylsiloxane-divinylbenzene (65 microm). The main parameters affecting the SPME process such as pH, salt additives, methanol content, memory effect, stirring rate and adsorption-time profile were studied. The method was developed using spiked natural waters such as ground water, sea water, river water and lake water in a concentration range of 0.1-10 microg/l. Limits of detection of studied compounds were determined in the range of 1-60 ng/l, by using electron-capture and mass spectrometric detectors. The recoveries of all fungicides were in relatively high levels (70.0-124.4%) and the average R2 values of the calibration curves were above 0.990 for all the analytes. The SPME conditions were finally optimized in order to obtain the maximum sensitivity. The potential of the proposed method was realized by applying it to the trace-level screening determination of fungicides and antifouling compounds in sea water samples originating from various Greek marinas. PMID- 11043596 TI - High-performance polyethylene glycol-coated solid-phase microextraction fibers using sol-gel technology. AB - The sol-gel method is applied for the preparation of solid-phase microextraction (SPME) fibers. An electron microscopy experiment suggested a porous structure for Superox-4 (polyethylene glycol, PEG) coating. SPME-GC analyses provided evidence that the sol-gel fibers have some advantages, such as high velocities of mass transfer, efficient extraction rates. high thermal stability, long life span, and spacious range of application for both polar and non-polar analytes. Efficient SPME-GC analyses of benzene-toluene-ethylbenzene-xylenes, phenols, phthalic diesters, naphthalene congeners and pesticides were achieved using sol-gel-coated PEG fibers. PMID- 11043597 TI - On-line monitoring trihalomethanes in chlorinated water by membrane introduction fast gas chromatography mass-spectrometry. AB - An analytical method based on membrane introduction and fast gas chromatography mass spectrometry (GC-MS) has been developed for the on-line monitoring of trihatomethanes (THMs) in chlorinated drinking water. The coupling of membrane introduction with fast GC-MS offers the advantage of membrane introduction as an on-line sampling device and fast GC-MS as a separation and identification method. While maintaining the on-line monitoring characteristic of traditional membrane introduction mass spectrometry (MIMS), the difficulty of distinguishing CHCl3 and CHBrCl2 in MIMS was overcome by rapid GC separation and MS analysis. Water permeated across the membrane affected the analysis of CHBr2Cl and CHBr3. A method based on controlling the injection temperature and injection time has been developed to overcome the moisture problem. This method is simple and less time consuming than the conventional moisture removing method. Under typical operating conditions, the sampling rate was about 20 samples h(-1) capable of on-line monitoring THMs in chlorinated drinking water. The detection limits of this system were found to be about 2 ppt, 4 ppt, 4 ppt, and 8 ppt for CHCl3 CHBrCl2, CHBr2Cl, and CHBr3, respectively. PMID- 11043598 TI - Enantiomeric separations of cationic and neutral compounds by capillary electrochromatography with monolithic chiral stationary phases of beta cyclodextrin-bonded negatively charged polyacrylamide gels. AB - Enantiomeric separation by capillary electrochromatography with beta-cyclodextrin bonded negatively charged polyacrylamide gels was examined. The columns used are capillaries filled with a negatively charged polyacrylamide gel, a so-called monolithic stationary phase, to which allyl carbamoylated beta-CD (AC-beta-CD) derivatives covalently bind. The capillary wall is activated first with a bifunctional reagent to make the resulting gel bind covalently to the inner surface of the fused-silica tubing. Enantiomeric separations of 15 cationic compounds were achieved using the above-mentioned columns and mobile phases of 200 mmol l(-1) Tris-300 mmol I(-1) boric acid buffer (pH 7.0 or 9.0) or 200 mmol l(-1) Tris-300 mmol l(-1) boric acid buffer (pH 7.0) containing an achiral crown ether (18-crown-6). Enantiomeric separations of two neutral compounds were also achieved using 200 mmol l(-1) Tris-300 mmol l(-1) boric acid buffer (pH 9.0) as a mobile phase. High efficiencies of up to 150,000 plates m(-1) were obtained. Both the within- and between-run reproducibilities of retention time and separation factor were good. The reproducibilities of retention time and separation factor for three different columns prepared from a different batch of monomers were acceptable. The gel-filled capillaries were stable for at least 3 months with intermittent use, utilizing the mobile phase of 200 mmol I(-1) Tris-300 mmol I( 1) boric acid buffer (pH 9.0). PMID- 11043599 TI - Determination of enantiomeric excess of alpha-hydroxy-3 phenoxybenzeneacetonitrile and its n-butyl ester by chiral high-performance liquid chromatography. AB - Determination of enantiomeric excess of alpha-hydroxy-3 phenoxybenzeneacetonitrile, an important intermediate in the production of several pyrethroid insecticides, is usually done after derivatization and gas chromatographic analysis on a beta-cyclodextrin-based column. In this communication we report a direct determination of enantiomeric excess of alpha hydroxy-3-phenoxybenzeneacetonitrile and its n-butyl ester by chiral HPLC on Chiralcel OJ (Daicel, Japan) in a single run without derivatization. PMID- 11043600 TI - Determination of sugar alcohols in confectioneries by high-performance liquid chromatography after nitrobenzoylation. AB - A method was developed for the determination of sugar alcohols, meso-erythritol, xylitol, D-glucitol, D-mannitol, maltitol and parachinit by high-performance liquid chromatography (HPLC). The sugar alcohols were converted into strong ultraviolet (UV)-absorbing derivatives with p-nitrobenzoyl chloride. HPLC was performed on a phenyl column, using acetonitrile-water (67:33) as mobile phase and UV detection (260 nm). The calibration curves for all sugar alcohols tested were linear in the 10-250 microg/ml range. The average recoveries of the sugar alcohols from four sugarless confectioneries spiked at 5 and 10% levels of six sugar alcohol standards ranged from 73.2 to 109.0% with relative standard deviations ranging from 0.7 to 9.0%. The detection limit of the developed method was 0.1% for the above sugar alcohols contained in the samples. PMID- 11043601 TI - Pressurised liquid extraction of ketones of polycyclic aromatic hydrocarbons from soil. AB - Pressurised liquid extraction (PLE) was used in the extraction of three ketones of polycyclic aromatic hydrocarbons from the sample of a soil highly contaminated with polycyclic polyaromatic compounds. The choice of solvent was the only factor that considerably influenced the extraction efficiency of PLE under the conditions recommended in Method 3545A promulgated by the United States Environmental Protection Agency. The dichloromethane-ethanol solvent mixture was found to be the most efficient solvent. PLE using this mixture provided better recoveries of all analysed ketones relative to Soxhlet extraction. PMID- 11043602 TI - Fatty acids and beta-carotene in australian purslane (Portulaca oleracea) varieties. AB - The fatty acid profile and beta-carotene content of a number of Australian varieties of purslane (Portulaca oleracea) were determined by GC and HPLC. The total fatty acid content ranged from 1.5 to 2.5 mg/g of fresh mass in leaves, 0.6 to 0.9 mg/g in stems and 80 to 170 mg/g in seeds. alpha-Linolenic acid (C18:3omega3) accounted for around 60% and 40% of the total fatty acid content in leaves and seeds, respectively. Longer-chain omega-3 fatty acids were not detected. The beta-carotene content ranged from 22 to 30 mg/g fresh mass in leaves. These results indicate that Australian purslane varieties are a rich source of alpha-linolenic acid and beta-carotene. PMID- 11043603 TI - UVB-induced hemolysis of rat erythrocytes: protective effect of procyanidins from grape seeds. AB - Besides erythema and sunburn reactions, UVB stress can promote erythrocyte extravasation from skin capillaries and hemolysis, and photosensitized hemoglobin can in turn lead to an overload of free radicals in dermis which exacerbates photodamage. The objective of this study was to investigate in rat erythrocytes (RBC) the pattern of events leading to membrane peroxidation and hemolysis following UVB insult (1.5-8.5 J/cm2), and the protective action of grape seed procyanidins. UVB causes a dramatic dose-dependent decrease of intracellular glutathione (paralleled by the formation of pro-oxidant ferryl-hemoglobin), of intramembrane vitamin E and of membrane fluidity, then a rise of conjugated dienes (CD), and thiobarbituric acid-reactive substances (TBARS) and finally a strong hemolytic effect. Procyanidins prevent membrane peroxidation (but not intracellular GSH depletion nor ferryl-hemoglobin formation), with a minimal effective concentration of 0.1 microM (IC50 for TBARS and CD after 120 min UVB exposure: 0.71 microM and 0.56 microM) and dose-dependently delay the onset of hemolysis, by 30 min at 0.1 mciroM, by 90 and 120 min at 0.5 and 1.0 microM. Epigallocatechin-3-O-gallate (EGCG) and catechin, typical constituents of the fraction, were significantly less potent. This since procyanidins (1 microM) inhibit the formation of phospholipid hydroperoxides of the inner (phosphatidylserine, phosphatidylethanolamine) and outer (phosphatidylcholine) layers of the RBC membrane (HPLC analysis), suppress the decrease in membrane fluidity due to lipid and protein thiol oxidation and spare vitamin E from consumption in a dose-dependent manner (0.1-1 microM). Hence procyanidins, preserving membrane phospholipids, since their strong antilipoperoxidant activity, may maintain in vivo the integrity of RBC in sub-epidermal capillaries and effectively counteract in dermis the onset/exacerbation of the UVB-induced skin photodamage. PMID- 11043604 TI - Phorbol ester-induced phosphoinositide hydrolysis in rat aorta: role of cyclooxygenase products. AB - This study investigates whether phorbol esters increase phosphoinositide hydrolysis in intact vascular smooth muscle, and the mechanism underlying the hydrolysis. Phorbol myristate acetate induced time- and concentration-dependent increases in phosphoinositide hydrolysis, as demonstrated by elevated inositol monophosphate levels, in deendothelialized rat aorta. The phorbol ester-elevated inositol monophosphate levels were abolished by indomethacin, a cyclooxygenase inhibitor, but were only partially decreased by SQ29548, a thromboxane A2/prostaglandin H2 receptor antagonist. SQ29548 also only partially decreased elevated inositol monophosphate levels due to prostaglandin E2, prostaglandin F2alpha, prostaglandin I2 and carbacyclin, a stable prostaglandin I2 analog. SQ29548 abolished elevated inositol monophosphate levels due to U46619, a stable thromboxane A2/prostaglandin H2 receptor agonist. These studies demonstrate that phorbol esters increase phosphoinositide hydrolysis in intact vascular smooth muscle, and that the increase is due, at lease in part, to endogenously released prostaglandins other than prostaglandin H2. PMID- 11043605 TI - Identification by differential display of the IF1 inhibitor peptide of ATP synthase/ATPase as a gene inducible in rat liver by pregnenolone 16alpha carbonitrile. AB - The synthetic steroid, pregnenolone-16alpha-carbonitrile (PCN), activates hepatic metabolism and elimination of xenobiotics mediated by its interaction with the PXR, a nuclear receptor that binds PCN and such glucocorticoids as dexamethasone (Dex). We used mRNA differential display to define further the domain of genes under the control of PCN/PXR. We found 76 cDNA fragments representing mRNAs differentially expressed in the liver of rats treated with PCN or Dex. Sequence analysis of one of these revealed a PCN induced cDNA fragment as IF1, an inhibitor peptide of ATP synthase/ATPase complex. Northern blot analysis revealed that IF1 was detectable in untreated liver and was induced 2-3 fold following treatment with PCN. IF1 mRNA was not detected in lung, heart, kidney, or testes of control or PCN treated rats. We conclude that IF1 inhibitor peptide is a novel representative of an apparently large set of previously unrecognized genes coordinately controlled by the PCN/PXR system to maintain homeostasis during toxic stress. PMID- 11043606 TI - Efficacy of muscarinic stimulation and mode of excitation-contraction coupling in bovine trachealis muscle. AB - We have compared the efficacy of cromakalim and nifedipine to inhibit acetylcholine (ACh) and pilocarpine-induced tonic contractions in control preparations and in tissues where a fraction of the muscarinic receptor population had been removed by alkylation with phenoxybenzamine (PBZ). Both agonists induced contractions by stimulating pharmacologically similar receptors, probably of the M3 muscarinic subtype. The receptor reserve was larger, and the coupling between stimulation and contraction (E-C coupling) more efficient when ACh was the stimulating agonist. For stimulations that produced equal levels of muscle response, cromakalim was more efficacious in inhibiting contractions induced by pilocarpine. The efficacy of cromakalim in relaxing contractions induced by ACh increased when the number of functional receptors decreased. Cromakalim and nifedipine decreased the efficiency of E-C coupling for ACh and pilocarpine. Cromakalim efficacy decreased in a sigmoid manner when stimulating concentrations of ACh (and receptor occupancy) increased, and there was an inverse relationship between receptor occupancy by ACh and cromakalim efficacy. In the presence of TEA, a K+ channel blocker, nifedipine almost completely inhibited contractions induced by the M3 muscarinic agonist bethanechol. These data indicate that in bovine tracheal smooth muscle, electro-mechanical coupling is an inherent part of muscarinic E-C coupling, but its functional expression is dependent upon the efficacy of stimulation. The data also suggest that the M3 receptor is coupled to a cellular pathway linked with the activation of K+ channels that exerts a potent functional antagonism against activation of voltage dependent Ca2+ entry. PMID- 11043607 TI - Distribution of [14C]suramin in tissues of male rats following a single intravenous dose. AB - PURPOSE: Suramin has been shown to have efficacy in treatment of prostate cancer. In the present study we evaluated distribution of [14C]suramin in tissues over time following a single intravenous dose. METHODS: Male rats were given a single IV dose of 300 mg/kg [14C]suramin and sacrificed at 1 or 6 hours, or at 1, 7, 14, 28, 56, or 84 days postdose. Radioactivity remaining in tissues was measured by quantitative whole body autoradiography. RESULTS: At one hour highest tissue activity was found in blood vessel walls and caecum, followed by lung, blood, skin, preputial, thyroid, brown fat, heart, kidney, lymph nodes, liver, salivary, adrenal, Harder's and lacrimal glands, prostate, and spleen. Considerable activity was present in membranes surrounding muscle groups, bone and other organs. Relatively low activity was found in brain tissue although persistent concentration was evident in choroid plexus. High levels were present in bladder and caecum contents. Activity declined in blood but continued to increase in many tissues at later time points. Kidney reached maximum levels at 7 days postdose and retained concentration considerably higher than other tissues over the course of the study. Concentrations in tissues were persistent and considerable activity remained at 84 days postdose. Terminal elimination half life in tissues was prolonged, approximately 39 days in blood and 91 and 102 days in kidney and spleen, respectively. Uptake in prostate was highest in membranous structures separating secretory lobules. CONCLUSION: Suramin is widely distributed to tissues and appears to have particular affinity for boundary membranes surrounding organs and other structural tissue elements, possibly due to uptake by glycosaminoglycans. Antitumor activity may be related to inhibition of growth factors associated with these elements. PMID- 11043608 TI - Decreased protein kinase C-epsilon expression in hypertrophied cardiac ventricles induced by triiodothyronine treatment in the rat. AB - To examine the effect of thyroid hormone-induced cardiac hypertrophy on PKC expression, changes in the expression of PKC isoforms were studied in hypertrophied cardiac ventricles induced by triiodothyronine (T3) injection in the rat. Injection with T3 for 8 days induced 49% increase in cardiac weight compared to controls. Immunoblot analysis of cardiac ventricular extracts showed the expression of PKC-delta, -epsilon, and -zeta in both control and T3-treated groups. The expression of PKC-epsilon decreased by 40% in hyperthyroid rat cardiac ventricles, while PKC-delta and -zeta expressions were barely affected. PKC-epsilon immunoreactivity decreased in both cytosol and membrane fractions. On the contrary, PKC-epsilon expression did not decrease in the extract of hypertrophied cardiac ventricles produced by aortic banding or aortocaval shunt. These results indicate that thyroid hormone down regulates PKC-epsilon expression in the hyperthyroid-mediated cardiac hypertrophy. PMID- 11043609 TI - The effect of Am-80, one of retinoids derivatives on experimental allergic encephalomyelitis in rats. AB - Experimental allergic encephalomyelitis (EAE) is an autoimmune model with inflammation and demyelination in the central nervous system, which resembles the human demyelinating disorder, multiple sclerosis (MS). In this study, we investigated the effect of Am-80, a synthetic retinoid, on EAE in DA rats. DA rats immunized with complete Freund's adjuvant (CFA) supplemented with myelin basic protein (MBP) developed severe EAE which reached the peak 12 to 14 days after immunization. Am-80 and prednisolone administered orally for 12 days after immunization diminished the clinical symptoms and infiltration of inflammatory cells in a dose dependent manner. However, after stopping administration, EAE recurred in DA rats treated with Am-80, but not with prednisolone. The different responses between Am-80 and prednisolone were not due to the difference in the tolerability to the MBP because both inhibited the delayed-type hypersensitivity response to MBP only during administration. To investigate the mechanism how Am 80 alone delayed the response, the expressional levels of mRNA for interleukin-6 (IL-6), interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) in spinal cord were examined. Transcriptional levels of IL-6, IFN-gamma and TNF alpha were parallel with the clinical symptoms of the disease in Am-80-treated rats, that is, expressional levels of their mRNA were diminished during the administration of Am-80, which then increased as soon as the administration was stopped. Among them, the expression of IL-6 mRNA was more rapidly and highly relapsed than that of the other two cytokines mRNA. However, prednisolone attenuated transcriptions of all these cytokines throughout the experiment. Therefore, these findings suggested that the inhibition of EAE is, in part, related to the inhibition of IL-6 production. However, there are many possible mechanism in the suppression of EAE by Am-80, further experiments will be necessary. PMID- 11043610 TI - Vitamin E, membrane fluidity, and blood pressure in hypertensive and normotensive rats. AB - Vitamin E treatment was found to lower blood pressure, and increase membrane fluidity in rats. The objectives of this study were to investigate the effects of the antioxidant, vitamin E, on the blood pressure and erythrocyte membrane fluidity in spontaneously hypertensive (SHR) and normotensive (WKY) rats. Membrane fluidity was assessed using spin labeling technique and electron paramagnetic resonance (EPR) spectroscopy. Two different spin labels were used in this study, 5-doxylstrearic acid (5-SASL) and 16-doxylstearic acid (16-SASL). The rats were given vitamin E, 3 days/week for 3 weeks and blood pressure was measured once weekly, using the tail-cuff method. Subsequently, blood was taken via heart puncture and erythrocytes were prepared for spin labeling. The fluidity of the membrane in the nonpolar region of erythrocytes from hypertensive rats was found quite different from that of normal rats as judged by the spectra of 16 SASL. The values of maximum splitting parameter of the EPR spectra of the spin label 5-SASL incorporated in erythrocyte membrane from both SHR and WKY rats, and the effects of vitamin E on membrane fluidity were compared. The maximum splitting parameter calculated from EPR spectra was larger for SHR than WKY rats. Additionally, the maximum splitting parameter calculated for vitamin E treated SHR and WKY rats were lower than those of their respective controls. As expected, the blood pressure of the SHR rats was found to be higher than that of the WKY rats. Vitamin E treated SHR and WKY rats showed significantly lower blood pressure than their controls. PMID- 11043611 TI - Gangliosides of rat eye lens: a severe reduction in the content of C-series gangliosides following streptozotocin treatment. AB - Gangliosides of eye lenses from normal and experimentally induced diabetic rats were investigated by methods including glycolipid-overlay techniques. Adult rat eye lens showed a complex ganglioside pattern that consisted of six major ganglioside components. These gangliosides were identified as GM3, GD3, GD1a, GD1b, GT1b, and GQ1b based upon their reactivity to anti-GM1 antibody after in situ sialidase treatment and mobility on thin-layer chromatography (TLC). Gangliosides in eye lens were further characterized by TLC-immunostaining with A2B5, a specific monoclonal antibody directed toward c-series gangliosides. Eye lens contained GT3 as the main c-series ganglioside component. Unexpectedly, the relative concentration of GT3 in total gangliosides of eye lens was highest among neural and extra-neural tissues examined. Administration of streptozotocin to rats caused a severe reduction in the GT3 content in eye lenses as early as day 3 without apparent changes in the composition of major gangliosides. Alloxan failed to produce such an effect despite producing similar hyperglycemic conditions. These results suggest that rat eye lens probably contains a streptozotocin susceptible cell type(s), which is highly enriched with c-series gangliosides. PMID- 11043612 TI - Excitation of nigral dopamine neurons by the GABA(A) receptor agonist muscimol is mediated via release of glutamate. AB - Previous electrophysiological studies have shown that the GABA(A)-receptor agonist muscimol is able to markedly increase the firing rate of rat nigral dopamine (DA) neurons. This action of the drug is paradoxical since local microiontophoretic application of the drug is associated with a clearcut inhibition of this neurons. In the present electrophysiological study, an attempt was made to analyze the mechanism of this action of the drug. Administration of muscimol (0.25-4.0 mg/kg, i.v.) was associated with a dose-dependent increase in firing rate as well as an increased bursting activity of the nigral DA neurons. Both these effects of muscimol were clearly antagonised by intravenous administration of the NMDA receptor antagonist MK 801(1 mg/kg) or by intracerebroventricular administration of the broad-spectrum excitatory amino acid receptor antagonist kynurenic acid. Furthermore, pretreatment with PNU 156561A (40 mg/kg, i.v., 5-8h), a compound that raised endogenous kynurenic acid levels about 9 times, also clearly antagonised the actions of muscimol. Indeed, this treatment reversed the excitatory action of muscimol into an inhibitory effect on the nigral DA neurons. Here, we report that the excitatory action of muscimol is mediated indirectly by release of glutamate. PMID- 11043613 TI - Vasorelaxing effects of Caesalpinia sappan involvement of endogenous nitric oxide. AB - Methanolic extract and two purified compounds (brazilin and hematoxylin) from Caesalpinia sappan were examined for their relaxant effects in isolated rat thoracic aorta. The methanolic extract significantly and dose-dependently relaxed the alpha1-receptor agonist phenylephrine-precontracted aortic rings, without affecting passive tension of these vessels. Removal of the vascular endothelium, inhibition of nitric oxide (NO) synthase with 0.1 mM Nomega-nitro-L-arginine and of cGMP biosynthesis with 10 microM methylene blue abolished the vasorelaxant effects of the herbal extract at doses up to 30 microg/ml. Similar vasorelaxant effects were observed with brazilin and hematoxylin. Therefore, these results suggest that brazilin and hematoxylin may be responsible for the vascular relaxant effects of C. sappan, via endogenous NO and subsequent cGMP formation. The vascular relaxant effects of the plant may contribute to its therapeutic actions. PMID- 11043614 TI - Helical CT scanning: the primary imaging modality for acute flank pain. AB - We set out to evaluate the accuracy of nonenhanced helical computed tomography (CT) scanning at stone detection in the patient with acute flank pain, and as a means of detecting noncalculus causes of acute flank pain. Between April 1995 and April 1997, 412 consecutive patients with acute flank pain underwent noncontrast enhanced helical CT. Two hundred eighty-one patients had confirmation of their CT diagnosis by other radiographic studies, urologic intervention, or spontaneous stone passage of calculi. We determined the presence or absence of urinary calculi, as well as the presence of other noncalculus pathology. CT scanning revealed a stone in 92/281 patients (32.7%) and no stone in 189/281 patients (67.3%). Of the 189 patients, 60/189 patients (32%) had another positive finding as a cause for flank pain. Eighty-one of 92 patients with a stone on CT (88%) had confirmation of stone disease by radiologic or surgical intervention. Eleven of 92 patients (12%) did not have confirmation of their diagnosis because of resolution of symptoms or refusal of further intervention. On helical CT scans 129/189 patients demonstrated no abnormalities. Two of 189 (1.5%) thought to be stone free by CT passed a stone. Helical CT had a sensitivity of 97%, a specificity of 92%, a positive predictive value of 88%, and a negative predictive value of 98% at stone detection. Noncontrast-enhanced helical CT is accurate and rapid in detecting calculus disease in patients with acute flank pain. Perhaps more importantly, it provides the added benefit of detecting noncalculus causes of flank pain in greater than 30% of patients. PMID- 11043615 TI - Prophylactic metoclopramide is unnecessary with intravenous analgesia in the ED. AB - Antiemetics are commonly prescribed as prophylaxis for nausea and vomiting when opiate analgesics are prescribed in the emergency department. This prospective, randomized, double-blind, placebo-controlled trial assessed the incidence of nausea and vomiting after morphine and pethidine (meperidine) analgesia, and the effect of metoclopramide on this incidence. Intravenous morphine or pethidine analgesia was administered with metoclopramide or placebo to 122 opiate-naive patients with acute severe pain. Seven patients (5.7%) experienced nausea, three in the metoclopramide group and four in the placebo group. One patient (0.8%) had vomiting. The frequency of other side effects was higher in the metoclopramide group (7.9% versus 3.4%). None of these differences reached statistical significance. The low incidence of nausea and vomiting after opiate analgesia, and higher incidence of side effects with metoclopramide, are consistent with controlled data in the literature. Prophylactic metoclopramide should not be used routinely in ED patients receiving parenteral morphine or pethidine analgesia. PMID- 11043616 TI - Initial fluid management of diabetic ketoacidosis in children. AB - The purpose of this study was to review the emergency department management of children presenting in diabetic ketoacidosis (DKA) to determine if current recommendations for fluid therapy are practiced. A 5-year retrospective chart review was conducted of all pediatric patients admitted with DKA to the University of Alberta Hospital. Presenting clinical and laboratory data, the initial fluid therapy, and insulin dose were analyzed. The therapy was also compared between sites of initial presentation (primary, secondary, or tertiary hospital). A total of 49 cases of DKA in 37 patients were reviewed. There were no significant clinical or biochemical differences between patients presenting at the three levels of hospital. Forty-one cases (84%) were given a saline bolus and the mean fluid volume given by 1 hour was 18.3 mL/kg. In the first hour 82% of patients presenting at a primary or secondary centre and 67% of those at the tertiary centre received more than 10 mL/kg. This excessive fluid therapy was also evident after 4 hours. Fluid management of children in DKA is excessive and not in keeping with current recommendations. Education of emergency physicians is needed to reduce fluid therapy and the risk of neurologic complications. PMID- 11043617 TI - Patient satisfaction with physician assistants (PAs) in an ED fast track. AB - The study objective was to determine patient satisfaction with physician assistants (PAs) in an emergency department (ED) fast track (FT). An additional goal was to determine if patients would be willing to wait longer to be seen primarily by an emergency physician (EP) rather than a PA. The study was conducted between March 1, 1999 and May 1, 1999 at a community hospital with an annual ED census of 48,644 patients; 18% are seen in the ED FT. All patients were seen primarily by a PA. An anonymous survey was given to patients at time of discharge. Patients rated their degree of satisfaction by placing an X on a 100 millimeter visual analogue scale. Patients also indicated if they would be willing to wait longer to be seen primarily by an EP rather than a PA. A total of 111 surveys were analyzed, for a response rate of 11%. Sixty-two patients (56%) were women and 49 men (44%), with a mean age of 28 years. Twenty-seven patients (24%) were younger than 18 years and required a legal guardian to complete the survey. The mean patient satisfaction score was 93 (95% CI: 90.27 to 95.73). Only 13 patients (12%) indicated they would be willing to wait longer to be seen primarily by an EP rather than a PA. Patients seen in an ED FT are very satisfied with the care rendered by a PA. Few patients would be willing to wait longer in such a setting to be seen primarily by an EP. PMID- 11043618 TI - Gender differences in state-wide EMS transports. AB - There are gender differences in emergency medical services (EMS) transports and management based on diagnosis. Data were extracted from the EMS State Ambulance Transport database. This database exists because of a legal requirement that all EMS transports generated by 911 calls and all interhospital transports be reported to the State EMS Bureau. All ambulance transports reported to the State EMS Division during 1995 were evaluated. Cases were excluded if they were aborted, admission or discharge transports, outpatient transports, or cases listed as "other" without a diagnosis. Gender-related treatment differences were determined for problems for which EMTs have specific treatment options. These were cardiac arrest, chest pain, allergic reactions, and extremity fractures. Results were compared using a two-tailed Chi squared or Fischer's Exact with significance at P < .05. Odds Ratios (OR) and 95% confidence intervals (CIs) were calculated. There were a total of 164,595 ambulance transports reported to the State EMS Division. Of these 76,074 (46%) were men and 88,521 (54%) were women. Of these, 50,211 were excluded. This left 52,607 injury transport and 61,777 illnesses transport. Men were significantly more likely than women to have injuries related to all-terrain vehicle accidents, motorcycle accidents, RV accidents, burns, gunshot wounds, and stab wounds. Men were significantly more likely than women to have illnesses related to cardiac arrest, dead on arrivals (DOAs), drowning, and smoke inhalation. For cardiac arrest transports, significantly more male patients presented ventricular fibrillation, more males received defibrillation, lidocaine, and bicarbonate, but more women received atropine. Male chest pain patients were more likely to receive oxygen and morphine and less likely to receive nitroglycerin. Male allergic reaction patients were more likely to receive an i.v. and subcutaneous epinephrine. Male extremity fracture patients were more likely to get an i.v. line, but there was no difference in morphine use or splinting. There are numerous disease-specific gender differences in the demographics of illness and injury transported by EMS. The use of various medications and procedures may also be related to gender. Understanding these differences may help in preparing EMS professionals for patient management. PMID- 11043619 TI - Experience with guidelines for cardiac monitoring after electrical injury in children. AB - The objective of the study was to evaluate the use of guidelines to determine the need of cardiac monitoring in children who sustained an electrical injury. The prospective use of guidelines since implementation July 1994 to June 1998 in a tertiary care pediatric teaching hospital was reviewed. Guidelines were cardiac monitoring for 24 hours is done on children with past cardiac history, loss of consciousness, voltage >240 volt or abnormal electrocardiogram (ECG); an ECG was obtained only when theoretical risk factors were present (tetany, decreased skin resistance by water or burns) or an unwitnessed event. Cardiac monitoring was performed in 29/224 (13%) patients (all normal) for 421 hours since implementation of the guidelines. Reasons included abnormal ECG (n = 10), voltage >240 volts (n = 6), lost of consciousness (n = 3), past cardiac history (n = 2), and unjustified (n = 9). There was no morbidity (0/172 patients 95% CI 0 to 1.7%) or mortality (0/224 patients 95% CI 0 to 1.3%). The guidelines were helpful in determining the need of cardiac monitoring in children after an electrical injury without any apparent risk. PMID- 11043620 TI - Male discomfort during the digital rectal examination: does examiner gender make a difference? AB - To determine if examiner gender affects men's perceived pain and embarrassment during an emergency department (ED) digital rectal examination, we prospectively studied male ED patients undergoing rectal examination. Each subject's perceived pain and embarrassment was gauged using 100 mm visual analog scales. Age, complaint, and examiner gender and training level were recorded. Two-tailed Mann Whitney or Kruskal-Wallis tests examined significant differences among group mean pain and embarrassment scores. Of 126 subjects, 60 had female and 66 had male examiners. Mean pain scores were similar for patients of female (36.5 mm) and male (37.4 mm) examiners (95% CI -9.8 to 11.5, P = .73). Mean embarrassment scores were similar for female (36.6 mm) and for male (32.9 mm) examiners (95% CI -7.9 to 15.3, P = .67). Younger male patients experienced more pain and embarrassment (P < .027). Examiner training level and prostatic examination did not affect the score. PMID- 11043621 TI - Ambulance use by high-acuity patients in a pediatric ED. AB - The objective of this study was to analyze ambulance usage by highest acuity patients as compared with all patients in a suburban pediatric hospital ED. A 1 year retrospective records analysis was conducted of all highest acuity patients (those patients triaged as emergent or critical or admitted to the intensive care unit). A total of 245 patients made 270 high-acuity visits to the ED in 1995. Thirty-one (13%) of the high-acuity patients arrived via ambulance; the rest arrived via private vehicle. The 31 high-acuity patients constituted 8% of the total number of patients arriving by ambulance. There was no significant difference in ambulance usage between insurance groups in the high-acuity patients. Only high-acuity patients with neurologic symptoms (primarily seizures) had a greater relative use of EMS transportation, with 39% of these patients arriving via ambulance (odds ratio 6.6, 95% confidence interval 2.6,16.6). High acuity patients account for the minority of total ambulance usage in our ED. PMID- 11043622 TI - Patient priorities with traumatic lacerations. AB - Clinical trials should use outcomes that are important to patients. We sought to determine the aspects of laceration management that are most important to patients. A prospective observational survey was conducted at one suburban and one urban university ED during November to December 1998 that included ED patients and visitors with and without current or prior lacerations. Trained research assistants approached 747 people of which 724 (97%) completed a 25-item closed question survey that evaluated demographics, prior laceration repairs, and assessed the relative importance of least painful repair, ED length of stay, cosmetic outcome, functional recovery, practitioner compassion, avoidance of wound infection, total costs, and missed days of work or school using a five-item Likert scale (not important-extremely important). Additionally, the relative importance of these items was compared. Data were analyzed with descriptive statistics and 95% confidence intervals (CIs). Seven hundred twenty-four people participated; 383 (53%) had prior lacerations; 92 (14%) had lacerations at the time of the survey. The most important aspect of care for respondents were normal function (28%), avoiding infection (20%), cosmetic outcome (17%), and least painful repair (17%). Based on Likert scale data, most important aspects of care were: avoiding wound infection (mean [95% CI], 4.58 [4.52 to 4.64]), normal function (4.54 [4.48 to 4.6]), cosmetic outcome (3.78 [3.68 to 3.88]), and least painful repair (3.84 [3.76 to 3.92]). Cost, length of stay, missed work/school, and compassion were less important (range, 3.0 to 3.7). Patients with facial lacerations chose cosmetic outcome as the most important aspect of care while all others chose function. Patients prioritize the medical outcomes of laceration repair (function, avoiding infection, cosmesis, pain) more than cost, compassion, ED length of stay and inconvenience (missed work/school). Cosmetic outcome is particularly important to patients with facial lacerations. This information should be useful when designing outcome studies of laceration management. PMID- 11043623 TI - Increasing frequency of dental patients in the urban ED. AB - This study examined the spectrum and frequency of dental disorders presenting to an urban ED. Data were retrospectively collected on all patients presenting with dental complaints between January 1, 1987 and December 31, 1995. Data included age, date and time of presentation, diagnosis, triage acuity and disposition. Of the 3,943 charts reviewed, 1,892 (48%) patients required emergent oral surgery management and 2,051 (52%) emergency physician management only. The frequency of dental patients increased from 4.4/1,000 total ED patients in 1987 to 11.5/1,000 in 1995 (P < .05). The rate of emergent dental trauma, emergent nontraumatic dental care, and nonemergent nontraumatic dental care increased similarly during the study period. There was no significant difference by day of week, but a significantly greater number of nonemergent patients presented between 7:00 a.m. and 3:00 p.m. (72%). The incidence of dental patients presenting to the ED increased significantly between 1987 and 1995. Approximately one-half did not require specialist consultation and could potentially have been managed by a primary care dental practitioner. PMID- 11043624 TI - Practicality and accuracy of prehospital rapid venous blood glucose determination. AB - Blood glucose testing plays an important role in emergency medicine. Although the use of visual reagent test strips is widely established in this setting, the accuracy of reflectometric blood glucose determinations under emergency conditions has rarely been investigated. In a prospective study, 522 of a total of 3,217 patients undergoing emergency blood glucose testing had parallel blood glucose measurements performed using a specific enzymatic method. These 522 patients (aged 61.4 years, 54% men, 90 cases of severe hypoglycemia) had an intravenous access placed at the scene of the emergency. Venous whole blood from the introducer needle of the access was applied to the test strip and the glucose measured with a GlucoTouch reflectometer (LifeScan, Inc.). A blood sample from the intravenous access was then immediately collected in a monovette for subsequent glucose determination in a chemical laboratory (hexokinase method) within 20 to 40 minutes. The emergency glucose measurements (mean: 7.3 mmol/L [95% confidence interval [CI] 6.9 to 7.7]; range: 0.55 to 27.7) correlated well with the reference laboratory results (Pearson's r = .98; linear regression analysis: slope 1.0, axial intercept 1.74). Error grid analysis also showed good agreement between corresponding measurements: zone A 96.7%, B 2.5%, C 0% and D 0.8%. The mean difference using the Bland-Altman method was 0.14 mmoVL; 2 SD 1.8 mmol/L; minimum -7.0 mmol/L; maximum 4.4 mmol/L. The accuracy of the rapid venous blood glucose determination by constantly changing emergency teams was high. Especially in 90 hypoglycemic patients, there were no deviations from the reference method that could have led to clinically relevant wrong decisions. The method of collecting whole blood directly from the venous access is simple and robust, and is independent of the hemodynamic status of the patient. PMID- 11043625 TI - The utility of maternal creatine kinase in the evaluation of ectopic pregnancy. AB - This investigation was designed to evaluate the utility of maternal creatine phosphokinase (CPK) in predicting the presence of an ectopic pregnancy (EP) in an emergency department (ED) setting. Twenty-one patients with the diagnosis of EP were randomly matched (1:1) with pregnant patients who subsequently ruled-out for EP. Serum CPK values at presentation were compared between the groups using two tailed ANOVA, odds ratio, and frequency tables were generated using our a priori hypothesis that a serum CPK of >70 mlU/dL may be useful as a predictor of EP. The mean serum CPK was 118mlU/dL in the EP group and 64 mlU/dL in the non-EP group (P < .0031). Controlling for age, race, and gestational age, there was an association between elevated serum CPK and EP in our study population (with an odds ratio of 6.5). The categorical evaluation (with 95% confidence interval [CI]) of CPK (>70 mIU/dL) as a predictor of EP follows: sensitivity - 100% (80.8 to 100); specificity-- 61.9% (38.7 to 81); PV(+) - 72.4% (52.5 to 88.6); PV(-) - 100% (71.7 to 100). We therefore conclude that a CPK level >70 mIU/dL may serve as an important adjuvant diagnostic tool in ruling-out EP. PMID- 11043626 TI - A rapid protocol to identify and exclude acute myocardial infarction: continuous 12-lead ECG monitoring with 2-hour delta CK-MB. AB - A prospective observational study was performed in 706 chest pain patients who underwent our chest pain evaluation protocol which consists of continuous 12-lead ST-segment monitoring with automated serial ECG (SECG) and a 2-hour delta (delta) CK-MB level determination before ED physician making final disposition decision to determine the incremental value of our 2-hour protocol for identifying myocardial infarction (MI) as compared with the initial ECG in combination with a baseline CK-MB. The initial ECG was obtained on presentation and considered positive if it revealed injury or ischemia. SECGs were obtained at least every 10 minutes and considered positive if it revealed new injury or ischemia. The baseline CK-MB value was considered positive if it was > or =12 ng/mL and index > or =4%. ACK-MB was defined as a difference between the 2 hour and baseline CK-MB and was considered positive if the value was > or =+1.5 ng/mL. MI was defined as acute myocardial infarction (AMI) or recent AMI (ie, AMI patients presenting on falling curve of CK-MB). The incremental value of the 2 hour protocol (ie, SECG in conjunction with deltaCK-MB) was more sensitive for identification of MI than the baseline protocol (ie, initial ECG in conjunction with the baseline CK-MB) (94.0% versus 55.4%; P < .0001) and reliably both identified (+LR = 14.6) and excluded MI (-LR = 0.06). SECG monitoring in conjunction with the 2 hour deltaCK MB allows for early identification and exclusion of MI, and can assist the ED physician in making appropriate treatment and disposition decisions. PMID- 11043627 TI - Hypothermia and afterdrop following open water swimming: the Alcatraz/San Francisco Swim Study. AB - To determine whether or not participants in open water swim events experience hypothermia and afterdrop, rectal temperature was measured for up to 45 minutes in 11 subjects following the New Year's Day Alcatraz Swim. This event was held in open water (11.7 degrees C [53.0 degrees F]) in the San Francisco Bay, and participants did not wear wetsuits or other protective clothing. Biophysical parameters, including surfacelvolume ratio, body mass index, and percent body fat were measured before the swim, and statistical analysis was done to determine predictors of temperature decrease and afterdrop duration. Applying the American Heart Association definition of hypothermia (less than 36.0 C [96.8 degrees F]), hypothermia was seen in 5 of the 11 subjects. Using a more rigorous and traditional definition (less than 35.0 degrees C [95.0 degrees F]), hypothermia was seen in only one subject. Afterdrop, defined as continued cooling following removal from cold stress, was seen in 10 of the 11 subjects. Surface/volume ratio (S/V) and body mass index (BMI) predicted the lowest recorded temperatures (P < .05; r(S/V) = -.71, r(BMI) = .72) and afterdrop duration (P < .05; r(SN) = -.75, r(BMI) = .69). These results suggest that hypothermia and afterdrop can occur commonly after recreational open water swimming, and that participants should be observed for signs of temperature decrease following removal from cold stress. PMID- 11043628 TI - Arthropod poisons and the cardiovascular system. AB - Animal poisons induce myocardial damage after envenomation. The cardiac manifestations after black widow spider bite are rarely observed and their prognostic significance are not known. Hymenoptera induce anaphylactic manifestations commonly observed in adults with previous coronary artery disease although myocardial involvement is observed in a few without previous heart disease. The mechanism of cardiac involvement after scorpion envenomation has been elucidated, however after widow spider bite or hymenoptera stings the mechanism of myocardial damage of those without previous heart disease is not clear. PMID- 11043629 TI - Electrocardiographic manifestations of CNS events. AB - Certain intracranial events produce electrocardiographic abnormalities, most often involving the T wave with diffuse, deep inversions. The amplitude of the T wave inversion is impressive, approaching 15 mm in some cases. Morphologically, the T wave is asymmetric with a characteristic outward bulge in the ascending portion. In the setting of a CNS event, relatively minor degrees of ST segment elevation are also seen in leads with obviously abnormal T waves; the ST segment elevation frequently is less noticeable than the T wave changes and is usually less than 3 mm. The T wave inversions with associated ST segment elevation are most pronounced in the mid-precordial and lateral precordial leads; such findings are also noted to a less extent in the limb leads. Other electrocardiographic features associated with acute CNS injury include prominent U waves of either polarity and QT interval prolongation, often exceeding 60% of its normal value, as well as malignant forms of bradycardia and tachycardia. PMID- 11043630 TI - Electrocardiographic manifestations of hyperkalemia. AB - Hyperkalemia is one of the more common acute life-threatening metabolic emergencies seen in the emergency department. Early diagnosis and empiric treatment of hyperkalemia is dependent in many cases on the emergency physician's ability to recognize the electrocardiographic manifestations of hyperkalemia. The electrocardiographic manifestations commonly include peaked T-waves, widening of the QRS-complex, and other abnormalities of altered cardiac conduction. Peaked T waves in the precordial leads are among the most common and the most frequently recognized findings on the electrocardiogram. Other "classic" electrocardiographic findings in patients with hyperkalemia include prolongation of the PR interval, flattening or absence of the P-wave, widening of the QRS complex, and a "sine-wave" appearance at severely elevated levels. A thorough knowledge of these findings is imperative for rapid diagnosis and treatment of hyperkalemia. PMID- 11043631 TI - ED identification of cardiac septal abscess using conduction block on ECG. AB - A case of cardiac septal abscess in a patient with a porcine bioprosthetic aortic valve who gradually developed a complete atrioventricular block on successive electrocardiograms (ECG) is reported. Emergency physicians should consider endocarditis with septal abscess in a patient with a prosthetic heart valve who presents with fever and a new conduction defect on ECG. PMID- 11043632 TI - Benign acute myositis. PMID- 11043634 TI - Embedded fishhook removal. PMID- 11043633 TI - Heliox treatment of severe croup. PMID- 11043635 TI - Sedation and local anesthesia preferences of emergency physician parents. PMID- 11043637 TI - Nonsurgical treatment of thyroid injury after blunt cervical trauma. PMID- 11043636 TI - High-dose verapamil-trandolapril induced rhabdomyolysis and acute renal failure. PMID- 11043638 TI - Tracking clinical outcomes of hospitalized patients. PMID- 11043639 TI - Intussusception in twins. PMID- 11043640 TI - Troponin I as a marker of cardiac toxicity in acute colchicine overdose. PMID- 11043641 TI - Ocular injuries from paintball. PMID- 11043642 TI - Time as responsible of ineligibility for thrombolytic treatment in stroke patients. PMID- 11043643 TI - Outbreak of Rift Valley fever--Saudi Arabia, August-October, 2000. AB - On September 10, 2000, the Ministry of Health (MOH), Kingdom of Saudi Arabia, and subsequently the Ministry of Health of Yemen received reports of unexplained hemorrhagic fever in humans and associated animal deaths from the southwestern border of Saudi Arabia and Yemen. Signs and symptoms of ill persons included low grade fever, abdominal pain, vomiting, diarrhea, jaundice with liver and renal dysfunction often progressing to disseminated intravascular coagulation, hepatorenal syndrome, and death. On September 15, using ELISA (antigen detection and IgM), polymerase chain reaction, virus isolation, and immunohistochemistry, CDC confirmed the diagnosis of Rift Valley fever (RVF) in all four serum samples submitted from Saudi Arabia. This report summarizes the preliminary results of the collaborative epidemiologic investigation performed by the Saudi Arabian MOH, CDC, and the National Institute of Virology, South Africa, of the first confirmed occurrence of RVF outside Africa. PMID- 11043644 TI - Measuring childhood asthma prevalence before and after the 1997 redesign of the National Health Interview Survey--United States. AB - Asthma is the most common chronic disease of childhood and a leading cause of disability among children (1,2). Since 1980, asthma prevalence has increased dramatically in children (3,4). The National Health Interview Survey (NHIS), the principal source of asthma prevalence data for the United States, was redesigned in 1997. This report presents NHIS data from 1980-1998 to examine the effect of the redesign on measuring trends in asthma prevalence overall and among age and racial subgroups of children. The findings indicate that although asthma prevalence estimates for 1997-1998 are lower than those preceding changes in the survey design, estimates after 1997 are not comparable to previous estimates. Additional data are needed to establish a new trend after 1997. PMID- 11043645 TI - Outbreak of Escherichia coli O157:H7 infection associated with eating fresh cheese curds--Wisconsin, June 1998. AB - On June 15, 1998, the Division of Public Health, Wisconsin Department of Health and Family Services, was notified of eight laboratory-confirmed and four suspected Escherichia coli O157:H7 infections among west-central Wisconsin residents who became ill during June 8-12. This report summarizes the outbreak investigation, which implicated fresh (held <60 days) cheese curds from a dairy plant as the source of infection. PMID- 11043646 TI - Enterovirus surveillance--United States, 1997-1999. AB - Enteroviruses accountfor an estimated 10-15 million symptomatic infections in the United States each year (1). At present, 66 serotypes of enteroviruses are recognized, including three poliovirus serotypes (2). A range of diseases is associated with nonpolio enterovirus infections, including aseptic meningitis, encephalitis, neonatal enteroviral disease, myocarditis, pericarditis, chronic infections among persons with compromised immune systems, poliomyelitis-like illness, hand-foot-and-mouth disease, nonspecific upper respiratory disease, and other manifestations (3). This report summarizes data from the National Enterovirus Surveillance System (NESS) and describes temporal trends of reported enterovirus infections in the United States during 1997-1999. PMID- 11043647 TI - Cocaine and metabolite elimination patterns in chronic cocaine users during cessation: plasma and saliva analysis. AB - Several reports suggest a prolonged elimination of cocaine and metabolites after chronic use compared with single or occasional use. This study was designed to measure the half-lives of cocaine in plasma and saliva of individuals who consumed cocaine on a frequent basis. The disposition and elimination patterns of cocaine and metabolites in the body fluids of chronic high-dose cocaine users during acute cessation of use were investigated. Plasma and saliva specimens were collected over a 12-h period during cessation and analyzed by gas chromatography mass spectrometry. Pharmacokinetic parameters were derived by noncompartmental analysis of plasma and saliva data. Results indicated a cocaine terminal T(1/2) of 3.8 h in plasma and 7.9 h in saliva. The terminal T(1/2) of benzoylecgonine was 6.6 h in plasma and 9.2 h in saliva. Compared with prior studies of acute low dose cocaine administration, these findings suggest that cocaine's half-life is longer in active street users than in occasional users though the half-life of its main metabolite benzoylecgonine remains similar (as do cocaine saliva-to plasma ratios). Thus, regular use of cocaine appears to alter the disposition and elimination of cocaine when compared to single or occasional use. PMID- 11043648 TI - Elimination of cocaine and metabolites in plasma, saliva, and urine following repeated oral administration to human volunteers. AB - Chronic administration of lipophilic drugs can result in accumulation and prolonged elimination during abstinence. It has been suggested that cocaine and/or metabolites can be detected in saliva and urine for an extended period following long-term, high-dose administration. The effects of chronic oral cocaine administration in healthy volunteer subjects with a history of cocaine abuse were investigated. Subjects were housed on a closed clinical ward and were administered oral cocaine in up to 16 daily sessions. In each session, volunteers received five equal doses of oral cocaine with 1 h between doses. Across sessions, cocaine was administered in ascending doses from an initial dose of 100 mg (500 mg/day) up to 400 mg (2 g/day), increasing by 25 mg/dose/session (125 mg/session). Participation in the study was terminated if cardiovascular safety parameters were exceeded. Plasma and saliva specimens were collected periodically during the dosing sessions and during the one-week withdrawal phase at the end of the study. All urine specimens were collected throughout the entire study. Specimens were analyzed for cocaine and metabolites by solid-phase extraction followed by gas chromatographic-mass spectrometric analysis in the SIM mode. The limit of detection for each analyte was approximately 1 ng/mL. The analytes measured included benzoylecgonine (BZE), ecgonine methyl ester, cocaine, benzoylnorecgonine, norcocaine, m- and p-hydroxycocaine, and m- and p hydroxybenzoylecgonine. Noncompartmental analysis was employed for the determination of plasma and saliva pharmacokinetic parameters. Urinary elimination half-lives for cocaine and metabolites were determined by constructing ARE (amount remaining to be excreted) plots. Two phases of urinary elimination of cocaine and metabolites were observed. An initial elimination phase was observed during withdrawal that was similar to the elimination pattern observed after acute dosing. The mean (N = 6) plasma, saliva, and urine cocaine elimination half-lives were 1.5 +/- 0.1 h, 1.2 +/- 0.2 h, and 4.1 +/- 0.9 h, respectively. For three subjects, the mean cocaine urinary elimination half-life for the terminal phase was 19.0 +/- 4.2 h. There was some difficulty in determining if a terminal elimination phase for cocaine was present for the remaining three subjects because of interference by high concentrations of BZE. A terminal elimination phase was also observed for cocaine metabolites with half life estimates ranging from 14.6 to 52.4 h. These terminal elimination half-lives greatly exceeded previous estimates from studies of acute cocaine administration. These data suggest that cocaine accumulates in the body with chronic use resulting in a prolonged terminal elimination phase for cocaine and metabolites. PMID- 11043649 TI - Cocaine and its major metabolites in plasma and urine samples from patients in an urban emergency medicine setting. AB - In this retrospective study, we examined the levels of cocaine and its major metabolites in plasma and urine from 29 randomly selected emergency department patients (19 males and 10 females, aged 19 to 55) whose urine screened positive for benzoylecgonine using fluorescence polarization immunoassay. Levels of cocaine along with benzoylecgonine, ecgonine methyl ester, and norcocaine were quantitated in EDTA plasma and urine from each patient using gas chromatography mass spectrometry with selected ion monitoring. Admission diagnosis and history were also obtained for each patient. In plasma, the levels were 16-130 ng/mL for cocaine (n = 3), 27-96 ng/mL for ecgonine methyl ester (n = 9), and 18-1390 ng/mL for benzoylecgonine (n = 22). Norcocaine was not detected in any of the plasma samples. In urine, the concentration ranges were 4-40,130 ng/mL for cocaine (n = 23), 36-660,500 ng/mL for ecgonine methyl ester (n = 27), and 9-2520 ng/mL for norcocaine (n = 9). All urine samples were positive for benzoylecgonine (106 3,361,000 ng/mL), and benzoylecgonine was the only metabolite present in two urine samples (at concentrations of 407 and 435 ng/mL). Two patients had plasma and urine samples positive for all analytes (except norcocaine in plasma). The patient with the highest urinary concentrations of cocaine (40,130 ng/mL), ecgonine methyl ester (660,500 ng/mL), benzoylecgonine (3,361,000 ng/mL), and norcocaine (2520 ng/mL) had a small quantity of benzoylecgonine (465 ng/mL) in plasma. No correlation was noted with patient history, admitting diagnosis or symptomatology, or plasma/urine levels of cocaine or any of its metabolites. PMID- 11043650 TI - Simultaneous GC-MS analysis of meta- and para-hydroxybenzoylecgonine and norbenzoylecgonine: a secondary method to corroborate cocaine ingestion using nonhydrolytic metabolites. AB - Positive benzoylecgonine (BZE) urinalysis results are sometimes challenged in legal and administrative proceedings on the grounds that the presence of BZE is due to the addition of cocaine to the urine sample with subsequent in vitro hydrolysis to BZE. Consequently, counsel for the respondent or defendant may move that an ecgonine methyl ester (EME) analysis be preformed because EME is presumed to be solely an in vivo cocaine metabolite. For these reasons, a sensitive and rapid gas chromatographic-mass spectrometric procedure was developed for the simultaneous analysis of m-hydroxybenzoylecgonine (m-OHBZE), p hydroxybenzoylecgonine (p-OHBZE), and N-desmethyl benzoyl ecgonine (norBZE), all of which are cocaine metabolites believed to arise exclusively via in vivo metabolism. Analysis of human urine specimens previously reported positive for BZE using GC-MS at the Department of Defense cutoff of 100 ng/mL demonstrated that at least one of the three metabolites was present in 79 of the 82 specimens studied (96.3%). Thus, the simultaneous analysis of r-OHBZE, p-OHBZE, and norBZE could be used to substantiate that the presence of BZE in urine specimens is the result of cocaine ingestion. Additionally, the premise that EME is a "true" in vivo cocaine metabolite was investigated by assessing the stability of cocaine in unpreserved urine samples at several pHs ranging from 5.0 to 9.0. PMID- 11043651 TI - Quantitation of cocaine, benzoylecgonine, cocaethylene, methylecgonine, and norcocaine in human hair by positive ion chemical ionization (PICI) gas chromatography-tandem mass spectrometry. AB - A total of 30 human head-hair samples were analyzed for cocaine (COC), cocaethylene (CE), benzoylecgonine (BE), methylecgonine (EME), and norcocaine (NCOC) using a sensitive positive ion chemical ionization gas chromatography tandem mass spectrometry (GC-MS-MS) method. All 30 hair samples had been previously submitted to the laboratory and had confirmed positive for cocaine. Hair samples (20 mg each) were cut into small segments (2-5 mm) and incubated overnight at 45 degrees C in 0.1 N HCl after the addition of 50 microL of an internal standard mix of COC-d3 (1.0 ng/mg), BE-d3 (0.5 ng/mg), EME-d3 (0.25 ng/mg), and NCOC-d3 (0.25 ng/mg). The samples were then extracted with Clean Screen extraction columns from United Chemical Technologies, Inc. The final extract was evaporated to dryness and derivatized with 50 microL of 1,1,1,3,3,3 hexafluoro-2-propanol and 50 microL of trifluoroacetic anhydride at 90 degrees C for 15 min. The derivatized samples were allowed to cool to room temperature, evaporated to dryness, and then reconstituted in 50 microL of ethyl acetate. Parent set masses (unbolded ions) and product ions were m/z 304 and m/z 182 and 82 (COC), m/z 307 and m/z 185 and 85 (COC-d3), m/z 318 and m/z 196 and 82 (CE), m/z 440 and m/z 318 and 105 (BE), m/z 443 and m/z 321 and 105 (BE-d3), m/z 296 and m/z 182, and 82 (EME), m/z 299 and m/z 185 and 85 (EME-d3), m/z 403 and m/z 386 and 105 (NCOC), m/z 406 and m/z 389 and 105 (NCOC-d3). Quantitation was accomplished by calculating the area ratio of the higher mass product ion (underlined ions) of analyte to the respective internal standard based on multilevel calibrations from 0.01 to 10.0 ng/mg. The GC-MS-MS method had a limit of detection of 0.01 ng/mg and a limit of quantitation of 0.05 ng/mg for all five analytes. COC, BE, and EME were detected in all 30 samples, and CE and NCOC were found in 19 and 29 samples, respectively. The average relative percentages of each metabolite normalized to the cocaine concentrations were 12.8%, 15.4%, 1.8%, and 2.5% for BE, CE, EME, and NCOC, respectively. PMID- 11043652 TI - The disposition of cocaine and opiate analytes in hair and fingernails of humans following cocaine and codeine administration. AB - This study investigated the disposition patterns of cocaine and opiates into hair and fingernail specimens collected from 8 volunteers enrolled in a 10-week inpatient clinical study. All subjects were African-American males with a confirmed drug use history. Scalp hair and fingernail scrapings were collected weekly throughout the course of the study. Head hair was collected from the posterior vertex region, and fingernail scrapings were collected along the entire ventral surface of the nail plate. The specimens were introduced to successive decontamination washes including an isopropanol wash and three phosphate buffer washes. All decontamination washes were collected and analyzed. All specimens were enzymatically digested prior to being subjected to solid-phase extraction and derivatization. Analyses were performed using electron impact gas chromatography-mass spectrometry. Analytes investigated included eight cocaine analytes and five codeine analytes. The limit of quantitation for all analytes ranged from 0.1 to 0.5 ng/mg for both matrices. Cocaine was present at the highest concentrations of any analyte in both hair and nail. Benzoylecgonine and ecgonine methyl ester were the primary metabolites in both matrices and were typically less than 15% of cocaine concentrations. Codeine was the only opiate analyte identified in either hair or nail. Observed drug disposition profiles were different for hair and nails. A significant dose-response relationship was observed for hair specimens. The mean peak concentrations in hair after low dosing were half the concentration observed after high-dose administration. Generally, no clear relationship was evident between nail drug concentrations and dose. Decontamination washes removed less than 20% of the total drug present in hair, but removed most of the drug concentrations (60-100%) in nail. This investigation demonstrated that higher concentrations of drug were found in the subjects' hair than in their fingernails and that cocaine was found in both matrices at a greater concentration than codeine. Although both hair and nail have similar physical and chemical properties and may share common mechanisms of drug incorporation, this clinical study suggests that there are distinct differences in their disposition profiles. PMID- 11043653 TI - Monitoring opiate use in substance abuse treatment patients with sweat and urine drug testing. AB - Although urine testing remains the standard for drug use monitoring, sweat testing for drugs of abuse is increasing, especially in criminal justice programs. One reason for this increase is sweat testing may widen the detection window compared to urine testing. Drug metabolites are rapidly excreted in urine limiting the window of detection of a single use to a few days. In contrast, sweat collection devices can be worn for longer periods of time. This study was designed to compare the efficacy of sweat testing versus urine testing for detecting drug use. Paired sweat patches that were applied and removed weekly on Tuesdays were compared to 3-5 consecutive urine specimens collected Mondays, Wednesdays, and Fridays (355 matched sweat and urine specimen sets) from 44 patients in a methadone-maintenance outpatient treatment program. All patches (N = 925) were extracted in 2.5 mL of solvent and analyzed by ELISA immunoassay for opiates (cutoff concentration 10 ng/mL). A subset (N = 389) of patches was analyzed by gas chromatography-mass spectrometry (GC-MS). Urine specimens (N = 1886) were subjected to qualitative analysis by EMIT (cutoff 300 ng/mL). Results were evaluated to (1) determine the identity and relative amounts of opiates in sweat; (2) assess replicability in duplicate patches; (3) compare ELISA and GC-MS results for opiates in sweat; and (4) compare the detection of opiate use by sweat and urine testing. Opiates were detected in 38.5% of the sweat patches with the ELISA screen. GC-MS analysis confirmed 83.4% of the screen-positive sweat patches for heroin, 6-acetylmorphine, morphine, and/or codeine (cutoff concentration 5 ng/mL) and 90.2% of the screen-negative patches. The sensitivity, specificity, and efficiency of ELISA opiate results as compared to GC-MS results in sweat were 96.7%, 72.2%, and 89.5%, respectively. Heroin and/or 6 acetylmorphine were detected in 78.1% of the GC-MS-positive sweat patches. Median concentrations of heroin, 6-acetylmorphine, morphine, and codeine in the positive sweat samples were 10.5, 13.6, 15.9, and 13.0 ng/mL, respectively. Agreement in paired sweat patch test results was 90.6% by ELISA analysis. For the purposes of this comparison of ELISA sweat patch to EMIT urine screening for opiates, the more commonly used urine test was considered to be the reference method. The sensitivity, specificity, and efficiency of sweat patch results to urine results for opiates were 68.6%, 86.1%, and 78.6%, respectively. There were 13.5% false negative and 7.9% false-positive sweat results as compared to urine tests. Analysis of sweat patches provides an alternate method for objectively monitoring drug use and provides an advantage over urine drug testing by extending drug detection times to one week or longer. In addition, identification of heroin and/or 6-acetylmorphine in sweat patches confirmed the use of heroin in 78.1% of the positive cases and differentiated illicit heroin use from possible ingestion of codeine or opiate-containing foods. However, the percentage of false-negative results, at least in this treatment population, indicates that weekly sweat testing may be less sensitive than thrice weekly urine testing in detecting opiate use. PMID- 11043654 TI - Detection times and analytical performance of commercial urine opiate immunoassays following heroin administration. AB - The Federal Workplace Drug Testing Program changed urine screening and confirmation cutoff concentrations for opiate testing from 300 to 2000 ng/mL in 1998. Morphine was the designated target compound. An additional heroin metabolite, 6-acetylmorphine (6-AM), was added to the testing procedure with a cutoff concentration > or = 10 ng/mL. Testing of 6-AM was required if morphine was positive to assist in medical review. A comparison of the new opiate cutoff concentrations was made with the older cutoff concentration at 300 ng/mL. Six commercial opiate immunoassays, four with a 300-ng/mL cutoff, ONLINE, EMIT, CEDIA and AxSym, and two with 2000-ng/mL cutoffs, ONLINE and EMIT, were selected to test 920 urine samples collected from 11 male human subjects following single doses of heroin. Eight received intravenous doses of 3, 6, and 12 mg heroin HCl and four smoked 3.5-, 5.2-, 10.5-, or 13.9-mg doses of heroin (base). In addition, 183 urine-based blind quality-control specimens were added to the study set to assess linearity, cross-reactivity, and interference. Total morphine, free morphine, and 6-AM were measured in each sample by gas chromatography-mass spectrometry (GC-MS). Linearity, cross-reactivity, and interference results for each immunoassay are described. Detection times, sensitivity, specificity, and efficiency of each assay were determined using data from the specimens collected after heroin administration. Detection times for morphine using the 300-ng/mL cutoff assays was approximately 12 h for low dose and 24 to 48 h for higher doses of heroin. For the two 2000-ng/mL cutoff concentration assays detection time was about 12 h. This was also the detection time for 6-AM by GC-MS. ONLINE had the lowest sensitivity, 60-74%, highest specificity, 98.8-100%, and least interference from a selection of common over-the-counter drugs and opioids. Increasing the cutoff to 2000 ng/mL from 300 ng/mL increased efficiencies of the assays from 72.7 to 82.6% to over 97%. PMID- 11043655 TI - Identification of hydrocodone in human urine following controlled codeine administration. AB - Allegations of illicit hydrocodone use have been made against individuals who were taking physician-prescribed oral codeine but denied hydrocodone use. Drug detection was based on positive urine opiate immunoassay results with subsequent confirmation of hydrocodone by gas chromatography-mass spectrometry (GC-MS). In these cases, low concentrations of hydrocodone (approximately 100 ng/mL) were detected in urine specimens containing high concentrations of codeine (> 5000 ng/mL). Although hydrocodone has been reported to be a minor metabolite of codeine in humans, there has been little study of this unusual metabolic pathway. We investigated the occurrence of hydrocodone excretion in urine specimens of subjects who were administered codeine. In a controlled study, two African American and three Caucasian male subjects were orally administered 60 mg/70 kg/day and 120 mg/70 kg/day of codeine sulfate on separate days. Urine specimens were collected prior to and for approximately 30-40 h following drug administration. In a second case study, a postoperative patient self-administered 960 mg/day (240 mg four times per day) of physician-prescribed oral codeine phosphate, and urine specimens were collected on the third day of the dosing regimen. Samples from both studies were extracted on copolymeric solid-phase columns and analyzed by GC-MS. In the controlled study, codeine was detected in the first post-drug-administration specimen from all subjects. Peak concentrations appeared at 2-5 h and ranged from 1475 to 61,695 ng/mL. Codeine was detected at concentrations above the 10-ng/mL limit of quantitation for the assay throughout the 40-h collection period. Hydrocodone was initially detected at 6-11 h following codeine administration and peaked at 10-18 h (32-135 ng/mL). Detection times for hydrocodone following oral codeine administration ranged from 6 h to the end of the collection period. Confirmation of hydrocodone in a urine specimen was always accompanied by codeine detection. Codeine and hydrocodone were detected in all specimens collected from the postoperative patient, and concentrations ranged from 2099 to 4020 and 47 to 129 ng/mL, respectively. Analyses of the codeine formulations administered to subjects revealed no hydrocodone present at the limit of detection of the assay (10 ng/mL). These data confirm that hydrocodone can be produced as a minor metabolite of codeine in humans and may be excreted in urine at concentrations as high as 11% of parent drug concentration. Consequently, the detection of minor amounts of hydrocodone in urine containing high concentrations of codeine should not be interpreted as evidence of hydrocodone abuse. PMID- 11043656 TI - The effects of collection methods on oral fluid codeine concentrations. AB - The use of a variety of alternative biological specimens such as oral fluid for the detection and quantitation of drugs has recently been the focus of considerable scientific research and evaluation. A disadvantage of drug testing using alternative specimens is the lack of scientific literature describing the collection and analyses of these specimens and the limited literature about the pharmacokinetics and disposition of drugs in the specimen. Common methods of oral fluid collection are spitting, draining, suction, and collection on various types of absorbent swabs. The effect(s) of collection techniques on the resultant oral fluid drug concentration has not been thoroughly evaluated. Reported is a controlled clinical study (using codeine) that was designed to determine the effects of five collection techniques and devices on oral fluid codeine concentrations. The collection techniques were control (spitting), acidic stimulation, nonacidic stimulation, and use of either the Salivette or the Finger Collector (containing Accu-Sorb) oral fluid collection devices. Preliminary data were collected from two subjects using the Orasure device. The in vitro drug recovery was also evaluated for the Salivette and the Finger Collector devices. With the exception of a single time point, codeine concentrations in specimens collected by the control method (spitting) were consistently higher than concentrations in specimens collected by the other methods. The control collection concentrations averaged 3.6 times higher than concentrations in specimens collected by acidic stimulation and 1.3 to 2.0 higher than concentrations in specimens collected by nonacidic stimulation or collection using either the Salivette or the Finger Collector devices. When calculated using oral fluid codeine concentrations from the clinical study, the elimination rate constant, t(1/2), AUC and the peak oral fluid concentrations demonstrated device differences. The slope of the elimination curve for codeine using the acidic collection method exceeded that of the other four methods. As a result, the t(1/2) for the acidic method was significantly less than that of the control method (1.8 vs. 3.0 h, respectively). Oral contamination contributed to the control method having higher AUC than that calculated using the other methods. There was considerable variation in peak codeine concentrations between devices and between individuals within each collection method. When samples were collected simultaneously with the Salivette and the Finger Collector, the mean codeine concentrations were similar. We were able to recover > or = 500 microL of oral fluid from 81.8% of the clinical samples collected with the Salivette. However, we were able to recover this volume from only 25.5% of the samples collected with the Finger Collector. In addition, the in vitro drug recoveries were lower using the Finger Collector. When oral fluid was collected nearly simultaneously by the control method and by use of the Salivette, mean control codeine concentrations were 2.3 times higher, but the duration of detection was similar for both methods. PMID- 11043657 TI - Liquid chromatography-electrospray ionization mass spectrometry for the detection of lysergide and a major metabolite, 2-oxo-3-hydroxy-LSD, in urine and blood. AB - A method is presented for the quantitative measurement of lysergide (LSD) and its metabolite 2-oxo-3-hydroxy-LSD (O-H-LSD) in urine and blood. O-H-LSD has been reported to have urinary concentrations many times higher than LSD. Measuring its presence in urine would significantly extend the detection time for confirming LSD abuse. A single-step liquid-liquid extraction was performed on 5-mL urine samples prior to separation by gradient liquid chromatography (LC). Electrospray ionization was used to produce the positively charged ions of O-H-LSD, 2-oxo-3 hydroxy-LAMPA (O-H-LAMPA, internal standard), LSD, and iso-LSD. Varying the orifice voltage in the intermediate-pressure region of the source generated the fragmentation necessary to produce qualifying ions. Selected ion monitoring allowed detection limits of 400 pg/mL and 100 pg/mL for O-H-LSD and LSD, respectively. The method was linear for O-H-LSD from 400 to 8000 pg/mL and for LSD from 100 to 6000 pg/mL. LSD-positive samples (n = 9) analyzed by the liquid chromatography-mass spectrometry method were found to contain mean concentrations of 6378 pg/mL O-H-LSD (332-21371 pg/mL) and 844 pg/mL LSD (177-2456 pg/mL). O-H LSD urinary concentrations were between 0.9 and 19.8 times higher than LSD (mean = 10.2). Whole-blood samples were also analyzed following additional sample cleanup. LSD was measured in the blood samples, but no O-H-LSD was detected. Enzymatic hydrolysis was carried out on LSD-positive samples (n = 6) to evaluate the existence of conjugated O-H-LSD. Beta-glucuronidase from Helix pomatia was incubated with urine samples at 37 degrees C, pH 5.2 for 24 h. At an enzymatic activity of approximately 4000 units per milliliter of urine, no significant (p = 0.05) difference was seen between hydrolyzed and nonhydrolyzed samples suggesting an absence of O-H-LSD-glucuronic acid conjugation. PMID- 11043658 TI - Metabolism of lysergic acid diethylamide (LSD) to 2-oxo-3-hydroxy LSD (O-H-LSD) in human liver microsomes and cryopreserved human hepatocytes. AB - The metabolism of lysergic acid diethylamide (LSD) to 2-oxo-3-hydroxy lysergic acid diethylamide (O-H-LSD) was investigated in liver microsomes and cyropreserved hepatocytes from humans. Previous studies have demonstrated that O H-LSD is present in human urine at concentrations 16-43 times greater than LSD, the parent compound. Additionally, these studies have determined that O-H-LSD is not generated during the specimen extraction and analytical processes or due to parent compound degradation in aqueous urine samples. However, these studies have not been conclusive in demonstrating that O-H-LSD is uniquely produced during in vivo metabolism. Phase I drug metabolism was investigated by incubating human liver microsomes and cryopreserved human hepatocytes with LSD. The reaction was quenched at various time points, and the aliquots were extracted using liquid partitioning and analyzed by liquid chromatography-mass spectrometry. O-H-LSD was positively identified in all human liver microsomal and human hepatocyte fractions incubated with LSD. In addition, O-H-LSD was not detected in any microsomal or hepatocyte fraction not treated with LSD nor in LSD specimens devoid of microsomes or hepatocytes. This study provides definitive evidence that O-H-LSD is produced as a metabolic product following incubation of human liver microsomes and hepatocytes with LSD. PMID- 11043659 TI - Detection of cannabis in oral fluid (saliva) and forehead wipes (sweat) from impaired drivers. AB - Saliva and sweat have been presented as two alternative matrices for the establishment of drug abuse. The noninvasive collection of a saliva or sweat sample, which is relatively easy to perform and can be achieved under close supervision, is one of the most important benefits in a driving-under-the influence situation. Moreover, the presence of certain analytes in saliva is a better indication of recent use than when the drug is detected in urine, so there is a higher probability that the subject is experiencing pharmacological effects at the time of sampling. We developed an original procedure using gas chromatography-mass spectrometry to test for delta9-tetrahydrocannabinol (THC), the psychoactive ingredient of cannabis, in oral fluid and forehead wipes, collected with Sarstedt Salivettes and cosmetic pads, respectively. Blood, urine, oral fluid, and forehead wipes were simultaneously collected from 198 injured drivers admitted to an Emergency Hospital in Strasbourg, France. Of the 22 subjects positive for 11-nor-9-carboxy-THC (THCCOOH) in urine, 14 and 16 were positive for THC in oral fluid (1 to 103 ng/Salivette) and forehead wipe (4 to 152 ng/pad), respectively. 11-Hydroxy-THC and THCCOOH were not detected in these body fluids. Two main limitations of saliva and sweat are apparent: the amount of matrix collected is smaller when compared to urine, and the levels of drugs are higher in urine than in saliva and sweat. A current limitation in the use of these specimens for roadside testing is the absence of a suitable immunoassay that detects the parent compound in sufficiently low concentrations. PMID- 11043660 TI - Consumption and quantitation of delta9-tetrahydrocannabinol in commercially available hemp seed oil products. AB - There has been a recent and significant increase in the use and availability of hemp seed oil products. These products are being marketed as a healthy source of essential omega fatty acids when taken orally. Although the health aspects of these oils is open to debate, the probability that oils derived from the hemp seed will contain delta9-tetrahyrdocannabinol (THC) is noteworthy. Recent additions to the literature cite a number of studies illustrating that the ingestion of these products results in urinary levels of the THC metabolite, delta9-tetrahyrdocannabinol carboxylic acid (THCA), well above the administrative cutoff (50 ng/mL) used during random drug screens. Testing performed by our laboratory found that the concentration of THC in hemp oil products has been reduced considerably since the publication of earlier studies. The purpose of this study is to quantitate the THC levels in commercially available hemp oils and to administer those oils tested to THC-free volunteers to determine urine metabolite levels following several 15-g doses. Two extraction protocols were evaluated for removing THC from the oil matrix: a single step liquid-liquid extraction was compared to a two-phase process using both liquid-liquid and solid phase techniques. Gas chromatography-mass spectrometry was used to determine THC levels in several products: four from Spectrum Essentials (3 bottled oils and 1-g capsules), two from Health from the Sun (1-g capsules and bottled oil) oils, and single samples of both Hempstead and Hempola hemp oils. These hemp oil products contained THC concentrations of 36.0, 36.4, 117.5, 79.5, 48.6, 45.7, 21.0, and 11.5 mg/g, respectively. The Abbott AxSYM FPIA and Roche On-Line KIMS immunoassays were used to screen the urine samples, and GC-MS was used to determine the amount of THC in each oil as well as confirm and quantitate THCA in the urine of study participants immediately before and 6 h after each dose. Peak THCA levels in the participants' urine ranged from 1 to 49 ng/mL. All volunteers were below positive screen and confirmation cutoffs within 48 h after cessation of ingestion. PMID- 11043661 TI - Loss of THCCOOH from urine specimens stored in polypropylene and polyethylene containers at different temperatures. AB - The loss of delta9-tetrahydrocannabinol (THCCOOH) from urine specimens stored in polypropylene and polyethylene containers at 4 degrees C and 25 degrees C was examined. All specimens were analyzed by GC-MS after sampling at various times over a one-week period. Data were analyzed by one-way analysis of variance and fitted with a first order kinetic equation. Rapid loss of THCCOOH was seen at 4 degrees C for both polypropylene (14% maximal loss, t(1/2) = 0.53 min) and polyethylene (17% maximal loss, t(1/2) = 5.77 min) bottles. At 25 degrees C, a small loss (< 5%) was observed in polypropylene and no significant loss was seen for urine in polyethylene. All losses stabilized within 1 h, and no further losses were seen over one week. The results indicate that THCCOOH binding may be due to decreased solubility of THCCOOH at lower temperatures and subsequent association of THCCOOH with the more lipophilic plastic. The results also indicate that polypropylene and polyethylene do not bind THCCOOH to such an extent as to compromise the integrity of specimens. PMID- 11043662 TI - Carbon monoxide stability in stored postmortem blood samples. AB - Carbon monoxide (CO) poisoning remains a common cause of both suicidal and accidental deaths in the United States. As a consequence, determination of the percent carboxyhemoglobin (%COHb) level in postmortem blood is a common analysis performed in toxicology laboratories. The blood specimens analyzed are generally preserved with either EDTA or sodium fluoride. Potentially problematic scenarios that may arise in conjunction with CO analysis are a first analysis or a reanalysis requested months or years after the initial toxicology testing is completed; both raise the issue of the stability of carboxyhemoglobin in stored postmortem blood specimens. A study was conducted at the Bexar County Medical Examiner's Office to evaluate the stability of CO in blood samples collected in red-, gray-, and purple-top tubes by comparing results obtained at the time of the autopsy and after two years of storage at 3 degrees C using either an IL 282 or 682 CO-Oximeter. The results from this study suggest that carboxyhemoglobin is stable in blood specimens collected in vacutainer tubes, with or without preservative, and stored refrigerated for up to two years. PMID- 11043663 TI - The characterization of human urine for specimen validity determination in workplace drug testing: a review. AB - One challenge facing the laboratory forensic toxicologist today is verifying the validity of the random urine specimen submitted for workplace drugs of abuse analysis. Determining whether urine substitution has occurred is best accomplished through the inspection of the specimen's appearance and the performance of specific laboratory tests, such as determining the concentration of biochemical metabolic waste products and measuring indices of urine concentration. Criteria for classifying submitted urine as substituted are postulated after an extensive review of the published scientific literature. Relevant studies that were evaluated include normal random urine reference interval studies, clinical studies involving the analysis of random urine specimens, theoretical dilutional limits, medical conditions resulting in overhydration, and water-loading studies. After compilation of the study data, derived substituted criteria of urinary creatinine < or = 5.0 mg/dL and urinary specific gravity < or = 1.001 are suggested. A urine specimen meeting these criteria may be considered substituted because it is not consistent with the clinical characteristics associated with normal human urine. PMID- 11043664 TI - Performance evaluation of four on-site drug-testing devices for detection of drugs of abuse in urine. AB - On-site drug tests are becoming increasingly more popular because of their easy test protocols and instantaneous results. This study evaluates the performance of four on-site drug testing devices that use competitive binding immunoassays to qualitatively determine the presence of drugs in urine: Triage Panel for Drugs of Abuse plus TCA, QuickScreen Pro-Multi Drug Screening Tests, Syva Rapid Test d.a.u. 5 and d.a.u. 2, and Rapid Drug Screen. All devices simultaneously determine the presence of the following drugs of abuse: amphetamine (AMP), benzoylecgonine (BE), 11-nor-9-carboxy-delta9-tetrahydrocannabinol (THCA), opiates (OPI), and phencyclidine (PCP). Triage and Rapid Drug Screen also simultaneously test for benzodiazepines (BZB) and barbiturates (BRB), whereas QuickScreen and Rapid Test require separate devices for the BZB and BRB analyses. Urine specimens (222) containing drug concentrations around or above cutoff values were screened by ONLINE or EMIT II immunoassays. Of these, 199 yielded positive gas chromatography-mass spectrometry results with at least 17 positive specimens in each drug class. Specimens with the target drugs added at 16.7% above and below the cutoff, 33.3% above and below the cutoff, and 66.7% above the cutoff were also used to evaluate the test devices. Sensitivity and specificity calculations demonstrated that Triage performed most predictably in the donor urine specimens and the drug-added specimens. In addition, it required the least amount of test volume and was the only device in which the appearance of a colored line indicated a positive result. Therefore, of the devices studied, Triage was the most dependable and reproducible on-site drug-screening device. PMID- 11043665 TI - Assessment of the ion-trap mass spectrometer for routine qualitative and quantitative analysis of drugs of abuse extracted from urine. AB - The ion-trap mass spectrometer (MS) has been available as a detector for gas chromatography (GC) for nearly two decades. However, it still occupies a minor role in forensic toxicology drug-testing laboratories. Quadrupole MS instruments make up the majority of GC detectors used in drug confirmation. This work addresses the use of these two MS detectors, comparing the ion ratio precision and quantitative accuracy for the analysis of different classes of abused drugs extracted from urine. Urine specimens were prepared at five concentrations each for amphetamine (AMP), methamphetamine (METH), benzoylecgonine (BZE), delta9 carboxy-tetrahydrocannabinol (delta9-THCCOOH), phencyclidine (PCP), morphine (MOR), codeine (COD), and 6-acetylmorphine (6-AM). Concentration ranges for AMP, METH, BZE, delta9-THCCOOH, PCP, MOR, COD, and 6-AM were 50-2500, 50-5000, 15-800, 1.5-65, 1-250, 500-32000, 250-21000, and 1.5-118 ng/mL, respectively. Sample extracts were injected into a GC-quadrupole MS operating in selected ion monitoring (SIM) mode and a GC-ion-trap MS operating in either selected ion storage (SIS) or full scan (FS) mode. Precision was assessed by the evaluation of five ion ratios for n = 15 injections at each concentration using a single-point calibration. Precision measurements for SIM ion ratios provided coefficients of variation (CV) between 2.6 and 9.8% for all drugs. By comparison, the SIS and FS data yielded CV ranges of 4.0-12.8% and 4.0-11.2%, respectively. The total ion ratio failure rates were 0.2% (SIM), 0.7% (SIS), and 1.2% (FS) for the eight drugs analyzed. Overall, the SIS mode produced stable, comparable mean ratios over the concentration ranges examined, but had greater variance within batch runs. Examination of postmortem and quality-control samples produced forensically accurate quantitation by SIS when compared to SIM. Furthermore, sensitivity of FS was equivalent to SIM for all compounds examined except for 6-AM. PMID- 11043666 TI - GC-MS analysis of methamphetamine impurities: reactivity of (+)- or (-) chloroephedrine and cis- or trans-1,2-dimethyl-3-phenylaziridine. AB - S-(+)-Methamphetamine is frequently found as the only isomer in urine specimens from methamphetamine abuseres. Enantiomerically pure S-(+)-methamphetamine can be synthesized from ephedrine or pseudoephedrine via chloroephedrine intermediates. These intermediates are unstable and capable of cyclizing to form cis- and trans 1,2-dimethyl-3-phenyl aziridine. Studies were done to determine if these intermediates could be detected when using a common gas chromatographic-mass spectrometric analytical method (derivatization with heptafluorobutyric anhydride, HFBA) for toxicological screening of methamphetamine. Analysis of (+)- or (-)-chloroephedrine after extraction into hexane and derivatization with HFBA indicated that both pseudoephedrine and ephedrine were the major compounds detected. Direct derivatization of a hexane solution of cis-1,2-dimethyl-3-phenyl aziridine yielded only the derivatives of ephedrine and pseudoephedrine, indicating that the aziridine intermediate is responsible for the formation of the ephedrine or pseudoephedrine. These studies indicate that the aziridine intermediates would not be detected in methamphetamine samples following HFBA derivatization. PMID- 11043667 TI - Differentiation of clobenzorex use from amphetamine abuse using the metabolite 4 hydroxyclobenzorex. AB - Clobenzorex (Asenlix) is an anorectic drug metabolized by the body to amphetamine, thus causing difficulty in the interpretation of amphetamine positive drug tests. Previous studies have shown the parent drug and several metabolites are excreted in urine. Clobenzorex itself has been detected for as long as 29 h postdose using a detection limit of 1 ng/mL. Despite this fact, several amphetamine-positive samples (> or = 500 ng/mL) contained no detectable clobenzorex. Thus, the absence of clobenzorex in the urine does not exclude the possibility of its use. To more definitively assess the possibility of clobenzorex use, evaluation of another metabolite was considered. One study reported the presence of unidentified hydroxy metabolites of clobenzorex for as long as amphetamine was detected in some subjects. To assess the viability of using a hydroxy metabolite to confirm the use of clobenzorex in samples containing amphetamine, 4-hydroxyclobenzorex was synthesized for this study. This metabolite proved to be easily detected and was typically found at levels higher than amphetamine in amphetamine-positive urines, long after clobenzorex itself was no longer detected. Samples obtained from a controlled single-dose study involving the administration of clobenzorex (30 mg) were analyzed for the presence of the 4-hydroxy metabolite. The analytical procedure used acid hydrolysis followed by liquid-liquid extraction and analysis with gas chromatography-mass spectrometry by monitoring ions at m/z 125, 330, and 364. 4 Hydroxyclobenzorex and its 3-Cl regioisomer were used in the identification and quantitation of the metabolite. Peak concentrations of 4-hydroxyclobenzorex were found at approximately 1:30-5:00 h postdose and ranged from approximately 5705 to 88,410 ng/mL. Most importantly, however, all samples that contained amphetamine at > or = 500 ng/mL also contained detectable amounts of this hydroxy metabolite (LOD 10 ng/mL), making it a valuable tool in differentiating use of clobenzorex from illicit amphetamine use. PMID- 11043668 TI - Quantitation of clonazepam and its major metabolite 7-aminoclonazepam in hair. AB - Clonazepam (CLO) is an anticonvulsant benzodiazepine approved by the Food and Drug Administration for use in the treatment of seizures. It produces pharmacological effects (depression, amnesia) similar to other compounds from the same therapeutic class, and in combination with alcohol, its CNS-depressant action can be significantly potentiated. As with some other benzodiazepines, CLO is a drug possibly used in "date-rape" situations. A method using solid-phase extraction followed by a highly sensitive negative chemical ionization gas chromatography-mass spectrometry for the simultaneous quantitation of CLO and its major metabolite 7-aminoclonazepam (7-ACLO) in hair was developed and validated. The method has potential application to alleged drug-facilitated rape cases. To determine the feasibility of detecting 7-ACLO and CLO in hair, specimens were collected from 10 psychiatric patients treated with CLO, divided into 2-cm segments, and analyzed. Standard curves for 7-ACLO (1-1000 pg/mg) and CLO (10-400 pg/mg) had correlation coefficients of 0.998. All precision and accuracy values were within acceptable limits. 7-ACLO was present in measurable quantities (1.37 1267 pg/mg) in 9 out of 10 patient samples. CLO concentrations in hair were much lower (10.7-180 pg/mg). In 4 out of 10 cases, CLO was not detected in hair. Two patients who had never been treated with CLO before received a single 2-mg dose of the drug. Approximately three weeks later, hair samples were collected, and measurable quantities of 7-ACLO (4.8 pg/mg) were detected in the first segment (proximal) of one of those samples, and traces of the drug were present in the other sample. We concluded that the 7-ACLO is being deposited in hair in much higher quantities than the parent drug and remains there for extended periods of time. Our study also indicates that it is possible to detect 7-ACLO after a single dose of CLO as in the typical date-rape scenarios. PMID- 11043669 TI - Simultaneous detection and quantitation of diethylene glycol, ethylene glycol, and the toxic alcohols in serum using capillary column gas chromatography. AB - Determination of toxic glycols and alcohols in an emergency setting requires a rapid yet accurate and reliable method. To simultaneously determine diethylene glycol (DEG) along with ethylene glycol, methanol, isopropanol, acetone, and ethanol, we modified a previously developed gas chromatographic (GC) method. The system used a Hewlett-Packard 6890 GC with EPC, a Gooseneck splitless liner, and an Rtx-200 capillary column (30 m x 0.53-mm i.d., 3 mm). After serum samples were deproteinized using ultrafiltration (Millipore Ultrafree-MC), 1 mL of the protein free filtrate was manually injected into the GC. Internal standards for alcohols (and acetone) and glycols were n-propanol and 1,3-butanediol, respectively. All compounds eluted within 3.5 min (linear temperature gradient from 40 to 260 degrees C); total run time was 6.5 min. Limit of detection and linear range for all compounds were 1 or 2.5 mg/dL and 0-500 mg/dL, respectively. In addition, there was no interference from propionic acid, propylene glycol, and 2,3 butanediol. The modifications in the equipment and temperature program allowed increased resolution and thus, detection and reliable quantitation of DEG and other common toxic glycols and alcohols of clinical interest. PMID- 11043670 TI - Duragesic transdermal patch: postmortem tissue distribution of fentanyl in 25 cases. AB - Fentanyl is a potent, short-acting narcotic analgesic widely used as a surgical anesthetic and for the control of pain when administered in the form of a transdermal patch. The success of the patch can be attributed to fentanyl's low molecular weight and its highly lipophilic nature, which enables it to be readily absorbed through the skin and subsequently distributed throughout the body. Over the past three years, the Los Angeles County Coroner's Toxicology Laboratory has encountered 25 cases involving Duragesic patches (fentanyl), and their postmortem tissue distributions are presented here. The analysis of fentanyl from postmortem specimens (3-mL or g sample size) consisted of an n-butyl chloride basic extraction followed by identification and quantitation on a gas chromatograph mass spectrometer using the selected ion monitoring (SIM) mode. The fentanyl ions monitored were m/z 245, 146, and 189 and the internal standard, fentanyl-d5 ions, were m/z 250, 151, and 194 (quantitation ion underlined). The linear range of the assay was 1.67 microg/L to 500 microg/L with the limit of quantitation and detection of 1.67 microg/L. The postmortem tissue distribution ranges of fentanyl in the 25 fatalities were as follows: heart blood, 1.8-139 microg/L (23 cases); femoral blood, 3.1-43 microg/L (13 cases); vitreous, +<2.0-20 microg/L (4 cases); liver, 5.8-613 microg/kg (22 cases); bile, 3.5-262 microg/L (15 cases); urine, 2.9-895 microg/L (19 cases); gastric, 0-1200 microg total (17 cases); spleen, 7.8 79 microg/kg (3 cases); kidney, 11 microg/kg (1 case); and lung, 31 microg/kg (1 case). The age of the decedents in this study ranged from 19 to 84, with an average age of 46. The modes of death included 15 accidental, 5 natural, 3 suicidal, and 2 undetermined. The main objectives of this paper are to show the prevalence of fentanyl patches in our community and to aid the forensic toxicologist with the interpretation of postmortem fentanyl levels in casework. PMID- 11043671 TI - Lamotrigine distribution in two postmortem cases. AB - Lamotrigine (Lamictal) is a new anticonvulsant drug recently approved for use in the United States. Although a therapeutic range for lamotrigine has not been definitively established, a range of between 2 and 14 mg/L has been reported. Two cases are presented in which lamotrigine was identified in cases investigated by the Office of the Chief Medical Examiner, State of Maryland. Lamotrigine was identified by gas chromatography-nitrogen-phosphorus detection following an alkaline extraction. A DB-5 column provided analytical separation; no derivatization was required. Confirmation was achieved by full scan electron ionization gas chromatography-mass spectrometry. In Case 1, primidone (11 mg/L) and phenobarbital (5.5 mg/L) were found in the heart blood in addition to lamotrigine (8.3 mg/L); in Case 2, no drugs other than lamotrigine (52 mg/L) were detected in the heart blood. The peripheral blood concentration in Case 2 was 54 mg/L. The liver lamotrigine concentrations in the two cases were 41 and 220 mg/kg. The medical examiner ruled that the cause of death in Case 1 was seizure disorder and the manner of death was natural. In Case 2, the medical examiner ruled that the cause of death was lamotrigine intoxication and the manner of death was undetermined. PMID- 11043672 TI - Loxapine intoxication: case report and literature review. AB - Loxapine is a dibenzoxazepine tricyclic compound used to treat schizophrenia in the United States since 1976. Metabolism includes demethylation to its primary metabolite, amoxapine. There are few documented reports of the disposition of loxapine in deaths due to overdose. This report discusses the overdose suicide of a 69-year-old white female found dead in her home by her husband. A prescription for loxapine (50-mg capsules) was found near the body. An autopsy was performed and heart blood, bile, vitreous humor, and gastric contents were submitted for toxicological analysis. The blood specimen was subjected to comprehensive testing that included volatile analysis by headspace gas chromatography (GC); acidic/neutral and basic drug screening by GC; benzodiazepine screening by high performance liquid chromatography; opiate screening by modified immunoassay; and acetaminophen, salicylate, and ethchlorvynol screening by colorimetry. Loxapine and amoxapine were detected in the basic drug screen. No other drugs were detected in the case specimens. The respective concentrations of loxapine and amoxapine in each specimen were as follows: heart blood, 9.5 and 0.6 mg/L; bile, 28.8 and 4.7 mg/L; gastric, 278 mg/L and negative; and vitreous, 1.5 mg/L and negative. A review of the literature showed that the heart blood concentration of loxapine measured in this case was the highest reported to date. Based on the autopsy findings, patient history, and toxicology results, the cause of death was determined to be acute intoxication of loxapine and the manner, suicide. PMID- 11043673 TI - Fatal strychnine poisoning: application of gas chromatography and tandem mass spectrometry. AB - The history and toxicological findings in a case of suicidal fatal strychnine poisoning are presented along with a description of the analytical methods. Detection and quantitation of strychnine in body fluids and tissues was performed by gas chromatography (GC) with nitrogen-phosphorus detection, using organic extraction and calibration by a standard addition method. Strychnine concentrations in subclavian blood (1.82 mg/mL), inferior vena cava blood (3.32 mg/mL), urine (3.35 mg/mL), bile (11.4 mg/mL), liver (98.6 mg/kg), lung (12.3 mg/kg), spleen (11.8 mg/kg), brain (2.42 mg/kg), and skeletal muscle (2.32 mg/kg) were determined. Confirmation of strychnine in blood and tissue was performed by GC with detection by tandem ion-trap mass spectrometry (MS). GC-MS-MS analysis, employing electron ionization followed by unit mass resolution and collision induced dissociation of strychnine, resulted in confirmatory ions with mass-to charge ratios of 334 (parent ion), 319, 306, 277, 261, 246, 233, and 220. Additional confirmation was provided by GC-MS-MS-MS analysis of each confirmatory ion, revealing an ion fragmentation pathway consistent with the molecular structure of strychnine. The case demonstrates body tissue and fluid distribution of strychnine in a fatal poisoning and the application of tandem MS in medical examiner casework. PMID- 11043674 TI - An unusual multiple drug intoxication case involving citalopram. AB - A 47-year-old male with a history of drug abuse and suicide attempts was found dead at home. The death scene investigation showed evidence of cocaine abuse and multiple drug ingestion. Citralopram, a new selective serotonin reuptake inhibitor, cocaine, oxycodone, promethazine, propoxyphene, and norpropoxyphene were identified and quantitated in the postmortem samples by gas chromatography mass spectrometry. The concentration of citalopram in the femoral blood was 0.88 mg/L. The heart blood concentration was 1.16 mg/L. Femoral blood concentrations of the other drugs were as follows: cocaine, 0.03 mg/L; oxycodone, 0.06 mg/L; promethazine, 0.02 mg/L; propoxyphene, 0.02 mg/L; and norpropoxyphene, 0.07 mg/L. Other tissue samples were also analyzed. The concentrations of cocaine, oxycodone, promethazine, and propoxyphene in the blood, liver, brain, and gastric contents did not suggest an intentional overdose. However, the possibility of multiple drug interactions including citalopram was evident. In this case, the citalopram concentrations were consistent with those reported in fatal cases involving multiple drug use. Citalopram was present in urine at a concentration of 0.9 mg/L. PMID- 11043675 TI - Chiral high-performance liquid chromatographic analysis of fluoxetine and norfluoxetine in rabbit plasma, urine, and vitreous humor using an acetylated beta-cyclodextrin column. AB - Fluoxetine (Prozac) is a potent selective serotonin reuptake inhibitor used for the treatment of major depression. Both fluoxetine (F) and its demethylated metabolite, norfluoxetine (NF), are racemic. S-Fluoxetine (SF) and S norfluoxetine (SNF) are more potent inhibitors of serotonin reuptake than R fluoxetine (RF) and R-norfluoxetine (RNF). Quantitation of individual enantiomers may provide a greater understanding of pharmacokinetic properties. The objective of this study was to perform a limited chiral selectivity study using rabbit plasma, urine, and vitreous humor analyzed by a solid-phase extraction protocol and a newly developed chiral analysis with an acetylated beta-cyclodextrin (CD) column. Liquid chromatographic parameters for CD were as follows: a mobile phase composition of methanol/0.3% triethylamine buffer, pH 5.6, (30:70), a flow rate of 1 mL/min, detection at 214 nm, and a temperature of 40 degrees C. Elution order was SNF, SF, RNF, and RF with capacity factors of 6, 7, 8, and 9, respectively. The corresponding resolution factors were as follows: R1,2 = 0.8, R2,3 = 1.2, and R3,4 = 0.9. The conditions for solid-phase extraction were optimized for Varian Bond Elut Certify columns. Following sample application, the column was rinsed with water, acetic acid, and then with methanol. Drug enantiomers were eluted with methylene chloride, isopropanol, and ammonium hydroxide (78:20:2). After extract evaporation, the extract residue was reconstituted for high-performance liquid chromatographic analysis. To investigate chiral pharmacology, a biodistribution study was performed by administering 2 mg/kg of F to five rabbits. Blood, urine, and vitreous specimens were collected. Plasma samples collected 45 min postinjection showed nearly equal concentrations of RF and SE After 24 h, the only metabolite detected in plasma was RNF. Drugs were not detectable in vitreous humor. Urine concentrations of SNF, SF, RNF, and RF were 51, 76, 34, and 8 microg/L, respectively. The CD column along with the described extraction protocol may be used for a chiral selectivity study of fluoxetine. PMID- 11043676 TI - The analysis of methadone in nail clippings from patients in a methadone maintenance program. AB - This study offers an analytical scheme for methadone in fingernail clippings. Nail specimens (0.18-16.33 mg) were collected from 30 consenting adults participating in a methadone-maintenance program along with questionnaires regarding their drug-use histories. The nail clippings were stored in plastic bags and transferred to the laboratory for analysis. They were decontaminated by sonication for 15-min intervals successively in 0.1% sodium dodecyl sulfate, water (three times), and methanol (three times). The methanolic washes were collected and screened for methadone by enzyme immunoassay (EIA). Three washes were found sufficient to provide EIA negative results. The decontaminated nail clippings were hydrolyzed in 1M NaOH. Aliquots of the hydrolysates were screened for methadone by EIA and confirmed by gas chromatography-mass spectrometry (GC MS). The mean methadone concentrations in fingernail clippings determined by EIA and GC-MS were 32.8 and 26.9 ng/mg, respectively. Hydrolysates of the equivalent of 10 mg of blank nail clippings were spiked with known concentrations of methadone and analyzed by the developed procedures in order to determine extraction recoveries and limits of detection of the two techniques. Based on our results, fingernails appear to be a potentially useful biological specimen for the analysis of methadone and the monitoring of patient compliance to methadone maintenance programs. PMID- 11043677 TI - Rapid gas chromatographic procedure for the determination of topiramate in serum. AB - A rapid gas chromatographic method for the routine determination in serum of the new anticonvulsant drug topiramate (Topamax) (TOP) is described. The method involves extracting 0.50 mL of sample, previously adjusted to pH 9.5 with saturated borate buffer with ethyl acetate. One-microliter aliquots of the extract were injected into a 10-m x 0.53-mm i.d. x 0.5-microm 100% methyl silicone megabore capillary column connected to a nitrogen-phosphorus detector. The column temperature was initially at 170 degrees C for 0.1 min, then programmed at 10 degrees C/min to 240 degrees C, then 20 degrees C/min to 280 degrees C for 0.5 min. Under these conditions of the assay, the retention times of TOP and mepivicaine, internal standard, were 4.0 and 3.4 min, respectively. Quantitative determinations were performed with peak-height ratios of TOP to the internal standard. Calibration curves were linear from 2.5 to 150 mg/L TOP. The assay had a limit of quantitation of 2.5 mg/L. The overall within-run precision of the method yielded coefficients of variation (CV) of 3.9% at 10 mg/L (n = 10) and 3.1% at 100 mg/L (n = 10). The overall between-run precision calculated by three determinations on a single day for a week yielded CVs of 7.3% at 23 mg/L (n = 12) and 7.8% at 85 mg/L (n = 12). Common anticonvulsant and basic/neutral extractable drugs were found not to interfere with the assay. At present, no correlation has been demonstrated between trough plasma TOP concentrations and clinical efficacy. However, TOP values observed in our laboratory in serums from patients receiving adjunctive treatment for seizure disorders ranged from 2.5 to 35 mg/L. PMID- 11043678 TI - A fatality due to accidental PineSol ingestion. AB - The case history and toxicological findings of a fatal PineSol intoxication are presented. An 89-year-old white female with Alzheimer's disease accidentally drank PineSol and was subsequently brought to the hospital where she was pronounced dead on arrival. Significant autopsy findings included acute erosive gastritis. There appeared to be no aspiration of PineSol into the lungs. Isopropanol along with 1-alpha-terpineol are the two major toxic ingredients of PineSol. The toxicological screening and quantitiation of 1-alpha-terpineol in postmortem fluids was performed by gas chromatography-mass spectrometry using a simple one-step extraction. Postmortem blood, urine, and gastric levels of 1 alpha-terpineol were 11.2 mg/L, 5.76 mg/L, and 15.3 g/L, respectively. Postmortem blood, vitreous humor, urine, and gastric acetone concentrations were 25, 31, 33, and 28 mg/dL. Postmortem concentrations of isopropanol were less than 10 mg/dL in the blood, vitreous humor, urine, and gastric contents. The cause of death was ruled acute 1-alpha-terpineol intoxication due to accidental ingestion of PineSol, presumably caused by confusion related to Alzheimer's disease. PMID- 11043679 TI - Exotic virus spreads across Northeast; researchers worry about wildlife. PMID- 11043680 TI - Problems facing women veterinarians, past and present. PMID- 11043681 TI - Types of leadership in the veterinary community. PMID- 11043682 TI - What is your diagnosis? Dorsal subluxation of the talus with bony opacities associated with the distal portion of the tibia. PMID- 11043683 TI - Animal behavior case of the month. Aggression toward household members. PMID- 11043684 TI - Effects of a synthetic facial pheromone on behavior of cats. AB - OBJECTIVE: TO evaluate the effects of a synthetic feline facial pheromone (FFP) on behavior and food intake of healthy versus clinically ill cats. DESIGN: Original study. ANIMALS: 20 cats were used in each of 2 studies. In each study, 7 cats were considered healthy, and 13 cats were determined to be clinically ill. PROCEDURE: In study 1, cats were assigned either to exposure to FFP (treated group; 4 healthy, 6 ill cats) or to exposure to the vehicle (70% ethanol solution; control group; 3 healthy, 7 ill cats). Cats were placed in a cage containing a small cotton towel that had been sprayed with FFP or vehicle 30 minutes previously. Cats were then videotaped for 125 minutes, and food intake was measured during this period. Videotapes were scored at 5-minute intervals for various behaviors. In study 2, cats were categorized in 1 of 2 groups; group 1 (2 healthy, 8 ill cats) had a cat carrier placed in their cages, and group 2 (5 healthy, 5 ill cats) did not. All cats were exposed to FFP, and 24-hour food intake was measured. RESULTS: Differences between behaviors of healthy versus clinically ill cats were not identified. In the first study, significant increases in grooming and interest in food were found in cats exposed to FFP compared with vehicle. For all cats, significant positive correlations were detected between grooming and facial rubbing, walking and facial rubbing, interest in food and facial rubbing, eating and facial rubbing, grooming and interest in food, and grooming and eating. In the second study, 24-hour food intake was significantly greater in cats exposed to FFP and the cat carrier, compared with cats exposed to FFP alone. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that exposure to FFP may be useful to increase food intake of hospitalized cats. PMID- 11043685 TI - Effect of a bioflavonoid dietary supplement on acetaminophen-induced oxidative injury to feline erythrocytes. AB - OBJECTIVE: To determine the effect of a commercial bioflavonoid antioxidant on acetaminophen-induced oxidative injury to feline erythrocytes. DESIGN: Randomized controlled study. ANIMALS: 45 healthy age-matched cats. PROCEDURE: Cats were assigned to 3 experimental groups. Groups 1 and 3 received a bioflavonoid antioxidant (10 mg/d) orally for 2 weeks. Groups 2 and 3 received an oxidative challenge with acetaminophen (90 mg/kg [41 mg/lb] of body weight, PO) on day 7. Packed cell volume, percentage of erythrocytes with Heinz bodies, blood methemoglobin concentration, and blood reduced and oxidized glutathione concentrations were determined at various times during the 2-week study period. RESULTS: Adverse effects were not associated with bioflavonoid antioxidant administration alone. Acetaminophen administration resulted in a significant increase in methemoglobin concentration in groups 2 and 3; differences were not detected between these groups. Heinz body concentrations in groups 2 and 3 increased after acetaminophen administration; however, the increase in cats that received the antioxidant was significantly less than in group-2 cats. Total blood glutathione concentrations did not change significantly in groups 2 and 3 after acetaminophen administration; however, ratio of reduced to oxidized glutathione concentration increased significantly after administration in group-2 cats, compared with group-3 cats. CONCLUSIONS AND CLINICAL RELEVANCE: Oral administration of bioflavonoid antioxidants to cats at risk for oxidative stress may have a beneficial effect on their ability to resist oxidative injury to erythrocytes. PMID- 11043686 TI - Cerebellar degeneration in Old English Sheepdogs. AB - OBJECTIVE: To evaluate related and unrelated Old English Sheepdogs (OESD) by clinical examination, histologic evaluation, and pedigree analysis to determine whether cerebellar degeneration develops in this breed and whether there are genetic implications. DESIGN: Case study and pedigree analysis. ANIMALS: 24 clinically normal or affected OESD; brain tissue specimens from 25 unaffected or affected OESD. PROCEDURE: Twenty-four OESD that were chosen because of a family history of gait abnormalities were given physical and neurologic examinations to determine whether they had clinical signs of cerebellar degeneration. Tissue specimens from 25 brains of OESD were examined histologically. Nine OESD that were determined to have cerebellar degeneration histologically as well as 2 clinically affected littermates of the histologically confirmed affected OESD were included in the pedigree analysis. Standard statistical evaluation of pedigrees for hereditary conclusions was used. RESULTS: Twelve of the 24 OESD evaluated by neurologic examination had a progressive gait abnormality. Clinical signs of cerebellar degeneration typically started later in life in OESD, compared with description for other dog breeds, and progressed ore slowly. Results of pedigree analysis revealed that 11 of 49 dogs were affected in 9 litters, providing an affected-to-total ratio of 22.49%. CONCLUSIONS AND CLINICAL RELEVANCE: Results of our study indicate that a slowly progressing late-onset form of cerebellar degeneration develops in OESD, and the mode of inheritance is by an autosomal recessive gene. PMID- 11043687 TI - Concurrent disorders in dogs with diabetes mellitus: 221 cases (1993-1998). AB - OBJECTIVE: To characterize concurrent disorders in dogs with diabetes mellitus (DM). DESIGN: Retrospective study. ANIMALS: 221 dogs with DM. PROCEDURE: Medical records were reviewed, and clinical signs, physical examination findings, and results of clinicopathologic testing, urinalysis, aerobic bacterial culture of urine samples, coagulation testing, endocrine testing, histologic evaluation, diagnostic imaging, and necropsy were recorded. RESULTS: For most dogs, CBC results were normal. Common serum biochemical abnormalities included hypochloremia (127 dogs, 60%) and high alanine aminotransferase (163, 78%), aspartate aminotransferase (78, 71%), and alkaline phosphatase (188, 90%) activities. Venous pH and serum ionized calcium concentration were measured in 121 and 87 dogs, respectively, and were low in 56 (46%) and 41 (47%) dogs. Lipemia was observed in 92 (42%) dogs. Urine samples from 159 (72%) dogs were submitted for aerobic bacterial culture, and 34 (21%) yielded bacterial growth. Escherichia coli was the most commonly isolated organism. Thirty-six (16%) dogs had dermatitis or otitis. Hyperadrenocorticism was diagnosed in 51 (23%) dogs on the basis of clinical signs and results of a low-dose dexamethasone suppression test (41 dogs), an adrenocorticotropic hormone stimulation test (5), both tests (4), or histologic evaluation of necropsy specimens (1). Acute pancreatitis was diagnosed in 28 (13%) dogs. Eleven (5%) dogs had tumors for which a histologic diagnosis was obtained. Eight (4%) dogs were hypothyroid. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that dogs with diabetes mellitus may have many concurrent disorders. The most commonly identified concurrent disorders included hyperadrenocorticism, urinary tract infection, dermatitis, otitis, acute pancreatitis, neoplasia, and hypothyroidism. PMID- 11043688 TI - Analysis of risk factors for the development of equine protozoal myeloencephalitis in horses. AB - OBJECTIVE: To investigate risk factors for development of equine protozoal myeloencephalitis (EPM) in horses. DESIGN: Case-control study. ANIMALS: 251 horses admitted to The Ohio State University Veterinary Teaching Hospital from 1992 to 1995. PROCEDURE: On the basis of clinical signs of neurologic disease and detection of antibody to Sarcocystis neurona or S neurona DNA in cerebrospinal fluid, a diagnosis of EPM was made for 251 horses. Two contemporaneous series of control horses were selected from horses admitted to the hospital. One control series (n = 225) consisted of horses with diseases of the neurologic system other than EPM (neurologic control horses), and the other consisted of 251 horses admitted for reasons other than nervous system diseases (nonneurologic control horses). Data were obtained from hospital records and telephone conversations. Risk factors associated with disease status were analyzed, using multivariable logistic regression. RESULTS: Horses ranged from 1 day to 30 years old (mean +/- SD, 5.7 +/- 5.2 years). Risk factors associated with an increased risk of developing EPM included age, season of admission, prior diagnosis of EPM on the premises, opossums on premises, health events prior to admission, and racing or showing as a primary use. Factors associated with a reduced risk of developing EPM included protection of feed from wildlife and proximity of a creek or river to the premises where the horse resided. CONCLUSIONS AND CLINICAL RELEVANCE: Development of EPM was associated with a number of management-related factors that can be altered to decrease the risk for the disease. PMID- 11043689 TI - Evaluation of risk factors associated with clinical improvement and survival of horses with equine protozoal myeloencephalitis. AB - OBJECTIVE: To investigate risk factors for use in predicting clinical improvement and survival of horses with equine protozoal myeloencephalitis (EPM). DESIGN: Longitudinal epidemiologic study. ANIMALS: 251 horses with EPM. PROCEDURE: Between 1992 and 1995, 251 horses with EPM were admitted to our facility. A diagnosis of EPM was made on the basis of neurologic abnormalities and detection of antibody to Sarcocystis neurona or S neurona DNA in CSF. Data were obtained from hospital records and through telephone follow-up interviews. Factors associated with clinical improvement and survival were analyzed, using multivariable logistic regression. RESULTS: The likelihood of clinical improvement after diagnosis of EPM was lower in horses used for breeding and pleasure activities. Treatment for EPM increased the probability that a horse would have clinical improvement. The likelihood of survival among horses with EPM was lower among horses with more severe clinical signs and higher among horses that improved after EPM was diagnosed. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of horses with EPM is indicated in most situations; however, severity of clinical signs should be taken into consideration when making treatment decisions. Response to treatment is an important indicator of survival. PMID- 11043690 TI - Use of an external ring fixator for correction of an acquired angular limb deformity in a donkey. AB - Numerous techniques for surgical correction of angular limb deformities in horses involving an osteotomy of the affected bone and stabilization with an internal fixation device have been described. However, because the osteotomy typically has to be performed at the level of the physis, leaving little bone between the physis and the nearest joint, stabilizing the osteotomy by use of internal fixation devices may be difficult. In horses with severe chronic angular limb deformities, the amount of soft-tissue contracture may make it impossible to correct the deformity during a single procedure without causing stretch injuries to the adjacent tendons and neurovascular structures. Adjustable external ring fixators incorporating hinged rods on 1 side of the limb and an angular motor assembly on the other may be useful for treatment of severe chronic angular limb deformities in younger equids, because they allow for gradual correction of the deformity. PMID- 11043691 TI - Detection and isolation of coronavirus from feces of three herds of feedlot cattle during outbreaks of winter dysentery-like disease. AB - Clinical signs of a winter dysentery-like syndrome in 6- to 9-month-old cattle in 3 feedlots included acute onset of diarrhea with high morbidity and low mortality, respiratory tract problems that included dyspnea, coughing, and nasal discharge, and high rectal temperatures. Bovine coronavirus was detected by use of an ELISA and immune electron microscopy in fecal and nasal swab samples and by immunohistochemical analysis of intestinal sections collected from calves during necropsy. Bovine coronavirus should be considered in the differential diagnoses for diseases that cause acute onset of bloody diarrhea in feedlot cattle. PMID- 11043692 TI - Evaluation of an in-house centrifugal hematology analyzer for use in veterinary practice. AB - OBJECTIVE: To compare CBC results obtained by use of an in-house centrifugal analyzer with results of a reference method. DESIGN: Prospective study. SAMPLE POPULATION: Blood samples from 147 dogs, 42 cats, and 60 horses admitted to a veterinary teaching hospital and from 24 cows in a commercial dairy herd. PROCEDURE: Results obtained with the centrifugal analyzer were compared with results obtained with an electrical-impedance light-scatter hematology analyzer and manual differential cell counting (reference method). RESULTS: The centrifugal analyzer yielded error messages for 50 of 273 (18%) samples. Error messages were most common for samples with values outside established reference ranges. Correlation coefficients ranged from 0.80 to 0.99 for Hct, 0.55 to 0.90 for platelet count, 0.76 to 0.95 for total WBC count, and 0.63 (cattle) to 0.82 (cats) to 0.95 (dogs and horses) for granulocyte count. Coefficients for mononuclear cell (combined lymphocyte and monocyte) counts were 0.56, 0.65, 0.68, and 0.92 for cats, horses, dogs, and cattle, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that there was an excellent correlation between results of the centrifugal analyzer and results of the reference method only for Hct in feline, canine, and equine samples; WBC count in canine and equine samples; granulocyte count in canine and equine samples; and reticulocyte count in canine samples. However, an inability to identify abnormal cells, the high percentage of error messages, particularly for samples with abnormal WBC counts, and the wide confidence intervals precluded reliance on differential cell counts obtained with the centrifugal analyzer. PMID- 11043693 TI - A brief overview of the analgesic and immunologic effects of acupuncture in domestic animals. PMID- 11043694 TI - Gram values and intakes: correction. PMID- 11043695 TI - Zinc supplementation. PMID- 11043696 TI - The three Cs of MNT: coverage, codes, and compensation. PMID- 11043697 TI - Multicultural competence in dietetics and nutrition. PMID- 11043698 TI - Research and the clinical dietitian: perceptions of the research process and preferred routes to obtaining research skills. AB - The art and science of dietetics are rooted in research, yet clinical dietitians do not generally participate in the research process. Focus group methodology was used to identify clinical dietitians' perceptions about incorporating research into clinical practice and to identify potential barriers to participation in research. In a series of 9 focus group sessions with 50 registered dietitians and 3 dietetic interns, attendees recognized the importance that research findings play in their practice. Barriers such as lack of time and low comfort levels with initiating and conducting research were discussed, and possible solutions to these barriers were suggested. Collaboration with academic dietitians or with research mentors was viewed as a useful and realistic pathway to increased participation in the research process. According to participants, increased involvement in research could also be facilitated by a supportive administrative structure within the facility, clinical management that is supportive of the research process, and collaboration among members of the dietetics staff. The findings of this qualitative investigation indicate that the central tenet of successful integration of clinical practice and research is collaboration-within dietetics departments, across the strata of facilities, and in particular, between clinical and research-oriented registered dietitians. PMID- 11043699 TI - Prevalence of marked overweight and obesity in a multiethnic pediatric population: findings from the Child and Adolescent Trial for Cardiovascular Health (CATCH) study. AB - OBJECTIVE: Determine the prevalence of marked overweight and obesity among children in the Child and Adolescent Trial for Cardiovascular Health (CATCH), identify high risk groups, and compare findings to other recent studies. DESIGN: Cohort study. SUBJECTS/SETTING: Five thousand one hundred-six school children who were participants in CATCH at baseline (age approximately 9 years) during 1991 and 4,019 of those children who had follow-up data from 1994 (age approximately 1 years) available. METHODS: Body mass index (BMI), triceps and subscapular skinfolds, subscapular to triceps skinfold (S/T) ratio, and an estimate of body fat distribution from skinfolds was calculated. Findings were compared to population-based reference data from the First National Health and Nutrition Examination Survey, 1971 to 1973 (NHANES I), to data from the Bogalusa Heart Study, and to data from the Third National Health and Nutrition Examination Survey, 1988 to 1994 (NHANES III). RESULTS: Children in CATCH were markedly heavier and fatter than the NHANES I population and more comparable to the NHANES III population, especially those in the upper percentiles. The prevalence of obesity based on BMI and triceps skinfolds >95th percentile among CATCH children was higher in boys than in girls at both baseline (boys 9.1%, girls 8.6%) and follow-up (boys 11.7%, girls 7.2%). It was higher among African-Americans and Hispanics than whites for both sexes. S/T ratios did not differ appreciably from those observed in the NHANES I reference population, suggesting that body fat distribution was more stable over time than BMI and skinfolds. APPLICATIONS: Our findings support other recent reports that American children, especially African American and Hispanic children, are becoming heavier and fatter. Preventive measures are warranted, especially for high-risk youth. PMID- 11043700 TI - Five-year-old girls' ideas about dieting are predicted by their mothers' dieting. AB - OBJECTIVE: To explore 5-year-old girls' ideas, concepts, and beliefs about dieting. DESIGN: Girls were asked to define dieting, to describe the behaviors dieting comprised, and were queried about links between dieting, weight control, and body shape. Parents completed questionnaires addressing family health history, demographics, and issues related to food, dieting, and weight control. SUBJECTS/SETTING: Participants were 197 girls aged 5 years and their parents. All girls lived with both biological parents, and were without food allergies or chronic medical problems. STATISTICAL ANALYSES PERFORMED: For 5 open-ended questions related to dieting, girls were categorized as either having or not having ideas about dieting. These ideas, concepts, and beliefs were categorized, and logistic regression examined predictors of girls' ideas about dieting. RESULTS: Depending on the question, from 34% to 65% of girls aged 5 years had ideas about dieting. Compared to girls whose mothers did not diet, girls whose mothers reported current or recent dieting were more than twice as likely to have ideas about dieting, suggesting that mothers' dieting behavior is a source of young girls' ideas, concepts, and beliefs about dieting. Among mothers, more than 90% reported recent dieting, and most reported use of both health-promoting and health-compromising dieting behaviors. APPLICATIONS: Women should be informed that weight control attempts may influence their young daughters' emerging ideas, concepts, and beliefs about dieting. Mothers should be encouraged to use health promoting rather than health-compromising weight control strategies, not only for their own well being, but to reduce the likelihood that daughters will incorporate health-compromising dieting behaviors into their concepts, ideas, and beliefs about dieting. PMID- 11043701 TI - Profiles of a healthful diet and its relationship to biomarkers in a population sample from Mediterranean southern France. AB - OBJECTIVES: The failure of single-nutrient supplementation to prevent disease in intervention studies underlines the necessity to develop a holistic view of food intake. The objectives of this study were to devise a diet quality index (DQI) and identify biomarkers of multidimensional dietary behavior. DESIGN: A nutrition survey was conducted in Mediterranean southern France by means of a food frequency questionnaire. The DQI was based on current dietary recommendations for prevention of diet-related diseases such as cardiovascular disease and some cancers. A second DQI included tobacco use. STATISTICAL ANALYSES: performed Spearman rank correlations, cross-classifications and intraclass correlations were computed between the DQI and biomarkers. RESULTS: Of the 146 subjects, 10 had a healthful diet and 18 had a poor diet. Erythrocyte omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)-beta carotene, and vitamin E concentrations were lower and cholesterol concentrations were higher in the poor diet; the difference was significant for EPA and DHA and borderline significant for vitamin E. Significant correlation was found between the DQI and vitamin E (-0.12), EPA (-0.30), and DHA (-0.28), and beta carotene (-0.17) when tobacco use was considered, but not between the DQI and cholesterol. The correlation coefficient reached 0.58 (P0.01) for a composite index based on all biomarkers except cholesterol. CONCLUSIONS: Subjects with a beta carotene levels greater thanl micromol/L, vitamin E greater than 30 micromol/L and EPA greater than 0.65% and DHA greater than 4% of fatty acids in erythrocytes were likely to have a healthful diet. Each biomarker indicated the quality of diet, but correlation was higher with a composite index. PMID- 11043702 TI - Comparison of the effectiveness of a telephone 24-hour dietary recall method vs an in-person method among urban African-American women. AB - OBJECTIVE: To examine the comparative accuracy of telephone and in-person 24-hour dietary recall methods. SUBJECTS: One hundred eighty-five African-American females, aged 40 years and older, recruited from Sunday church services in Baltimore City, Md. METHODS: Participants were trained to estimate portion size with plastic food models and a 2-dimensional food recall booklet. Dietary intake was then assessed with 2 in-person 24-hour dietary recalls and 1 telephone 24 hour dietary recall, all using a computer-assisted, multiple pass approach. Results from the 2 in-person recalls were averaged and compared with the results from the telephone recall. STATISTICAL ANALYSES: Cross-tabulation, paired t test, Pearson's correlation, chance-corrected agreement, and stepwise linear regression analyses were performed. RESULTS: There were no significant differences between the telephone and in-person methods for any nutrient. Agreement between methods was moderate for all major dietary components, with corrected correlations between methods ranging from 0.26 to 0.97 (P<.001), and kappas ranging from 0.155 to 0.372 (P<.01). Levels of low-energy reporting were high (88% telephone, 91% in person), though there were no significant differences between methods. CONCLUSIONS: The telephone 24-hour dietary recall method appears to be comparable to the standard in-person method among older African-American women. Portion-size training in person may make subsequent telephone dietary recalls acceptable in this population. PMID- 11043703 TI - Model for multicultural nutrition counseling competencies. AB - A model for multicultural nutrition counseling competencies for registered dietitians was developed and tested. Six hundred four registered dietitians who were members of The American Dietetic Association Public Health Nutrition Practice Group or directors of dietetic internships and didactic programs in dietetics were selected by a stratified random sample method and were mailed a survey. Respondents rated each of 46 competencies using a Likert scale to delineate how essential each competency will be for entry-level dietitians in the next 10 years. Of the 60% who responded (n=363), 94.4% met the study selection criteria. Most were white (85.7%), spoke English as their primary language (96.8%), and had a master's degree (64.4%). Many (37.9%) worked in community/public health facilities or organizations, and 50.4% provided nutrition counseling or education to clients culturally different from themselves. Exploratory principal components analysis extracted 3 factors with 28 competencies loading on them: multicultural nutrition counseling skills, multicultural awareness, and multicultural food and nutrition knowledge. Subjects responded similarly whether or not they provided nutrition counseling to culturally different clients. Secondary analysis revealed no significant interaction or differences between how bilingual dietitians and those of color scored items in the 3 factors. The resulting model is a guideline that can be used by educators to enhance dietetics education and training and by public health nutritionists as a basis for self-evaluation and selection of continuing education opportunities to enhance their multicultural nutrition counseling competence. PMID- 11043704 TI - Designing a quality assurance system for dietary data in a multicenter clinical trial: Women's Intervention Nutrition Study. AB - Reliable dietary intake data are essential for determining outcomes in nutrition related clinical trials. Nevertheless, systems for quality assurance of dietary intake data are often slighted in the design of such trials and not incorporated or monitored as the trials continue. The Women's Intervention Nutrition Study (WINS), a multicenter clinical trial investigating the effect of reduction of dietary fat intake together with adjuvant systemic therapy on recurrence rates in and survival of postmenopausal women with early stage, surgically treated, breast cancer, has developed a quality assurance system to minimize errors and to produce data that are complete and reliable. The system involves development of standardized procedures for data collection, a quality control program to evaluate the data collected, and continual monitoring and reevaluation. The WINS system is offered as a model for studies collecting dietary intake data, no matter how simple or complex the trial design. PMID- 11043705 TI - Improving dietetics education with interactive communication technology. AB - Changes occurring in health care, education, and technology are altering dietetics education. A model of learnercentered, cooperative, distance education based on interactive online technology is described for use in a dietetic internship. Evaluation of this model includes review of key-feature exams, results of computer attitudes surveys, use of the technology by interns, exit interviews, and performance on the examination for registered dietitians. In a pilot study of the model with 8 subjects, comfort using the Internet improved significantly. Use of interactive communication technology in dietetics education has the potential to improve competency, technological aptitude, professional partnering skills, and lifelong learning skills. PMID- 11043706 TI - Benefits associated with serving as a preceptor for dietetic interns. PMID- 11043707 TI - Dietary habits and health beliefs of Korean-Americans in the San Francisco Bay Area. PMID- 11043708 TI - Accuracy of phylloquinone (vitamin K-1) data in 2 nutrient databases as determined by direct laboratory analysis of diets. PMID- 11043709 TI - Scope of practice for qualified dietetics professionals in diabetes care and education. PMID- 11043710 TI - Position of the American Dietetic Association: the impact of fluoride on health. AB - Fluoride is an important element for mineralization of body tissues. The use of topical and systemic fluoride for oral health has resulted in major reductions in dental caries and its associated disability. Fluoridation of public water supplies has been endorsed by over 90 professional health organizations as the most effective dental public health measure in existence. Still, about half of the US population fails to receive the maximum benefits possible from community water fluoridation and the use of fluoride products. Fluoride also plays a role in bone health. The role of high doses of fluoride for prevention of osteoporosis is undergoing active study and is considered experimental at this point. Dietetics professionals should routinely monitor and promote the use of systemic and topical fluorides, especially in children and adolescents. The American Dietetic Association strongly reaffirms its endorsement of the use of systemic and topical fluorides, including water fluoridation, at appropriate levels of intake, as an important public health measure. However, clients should be cautioned that experimental use of high intakes of fluoride should be avoided unless they are participating in clinical trials. PMID- 11043711 TI - Struma ovarii with hyperthyroidism. AB - A case of hyperthyroid disease after total thyroidectomy is reported. An initial scintigraph with Tc-99m sodium pertechnetate confirmed the absence of uptake in the neck. The view of the pelvis revealed a rounded area of increased uptake on the left, near the sacroiliac joint (results of bone scintigraphy were negative). A wholebody scan with iodine-131 confirmed a struma ovarii that was bilateral, which occurs in 5% to 10% of patients with this condition. PMID- 11043712 TI - Thyroid hemiagenesis. AB - Thyroid hemiagenesis is a rare congenital anomaly in which one thyroid lobe fails to develop. Tc-99m pertechnetate scintigraphy and ultrasonography can be used to visualize this anomaly. The authors evaluated four cases with Tc-99m pertechnetate scintigraphy and ultrasonography and confirmed thyroid hemiagenesis. In one case, a Tc-99m MIBI scan excluded the suppresssion of the contralateral thyroid lobe. Tc-99m MIBI can be used to confirm the diagnosis of thyroid hemiagenesis. PMID- 11043713 TI - Thyroid hemangioma: a case report with a review of the literature. AB - A 53-year-old man was found serendipitously to have increased blood-pool uptake in the right lobe of the thyroid during a radionuclide ventriculography study for left ventricular function. Primary thyroid hemangioma is a rare condition, with only two cases reported in the literature. Secondary hemangioma may occur as a result of fine-needle aspiration. This technique may cause hematoma formation, which generally resolves normally, but on rare occasions it can lead to cavernous hemangioma formation. In patients with a thyroid swelling who have a cold nodule on a thyroid scan and only blood on repeated fine-needle aspiration, Tc-99m erythrocyte blood-pool imaging may be performed to diagnose hemangioma. PMID- 11043714 TI - Multiple brown tumors simulating bone metastases: a case of parathyroid adenoma coexisting with papillary carcinoma of the thyroid. AB - A 55-year-old man with multiple brown tumors who initially was thought to have multiple bone metastases is described. He had elevated parathyroid hormone levels and was referred to the nuclear medicine department, where a parathyroid adenoma was diagnosed. At surgery, abnormal lymph nodes were seen, which were found to contain metastatic thyroid papillary carcinoma cells. On further exploration, the patient's bone scintigraph revealed multiple sites of increased uptake but no bone abnormalities on whole-body iodine and Tc-99m MIBI scans. PMID- 11043715 TI - Prognostic value of myocardial perfusion tomographic imaging in patients after percutaneous transluminal coronary angioplasty. AB - PURPOSE: The authors assessed the prognostic value of stress myocardial perfusion tomographic imaging (SPECT) in patients with recurrent angina or inconclusive results of exercise electrocardiographic tests after successful percutaneous transluminal coronary angioplasty (PTCA). MATERIALS AND METHODS: After PTCA, 70 patients (54 men, 16 women; 41 after myocardial infarction; mean age, 56 +/- 9 years) underwent TI-201 or Tc-99m sestamibi SPECT studies. SPECT patterns were divided into normal (n = 25), fixed defects (n = 15), and reversible or combined fixed plus reversible defects (n = 30). A cardiac event was defined as either cardiac death, nonfatal myocardial infarction, or unstable angina requiring further revascularization. RESULTS: During an average follow-up of 25 +/- 10 months, two patients had severe outcomes (one cardiac death and one nonfatal myocardial infarction), and revascularization was required in 13 patients. In patients with normal SPECT or fixed defects, the annual event rate was low (1.2%), with only one revascularization. In patients with reversible or combined defects, the annual event rate was significantly greater (22.4%; chi square = 17.32, P = 0.00003). CONCLUSIONS: Normal perfusion or fixed defects predict a benign prognosis in patients after successful PTCA. The presence of stress induced reversible defects appears to be the best predictor of future cardiac events. PMID- 11043716 TI - Late correlative imaging findings of previous acute infective spondylitis. AB - PURPOSE: To evaluate late correlative imaging findings in relation to clinical outcome in persons with previous acute infective spondylitis, the authors performed bone and leukocyte scintigraphy (planar and SPECT imaging) and magnetic resonance imaging of the spine in nine patients (two men, seven women; mean age, 66 years; age range, 57 to 84 years) 3.1 years (range, 0.9 to 6.2 years) after the acute disease. METHODS: All images were evaluated visually. The relevant uptake on SPECT images was also quantitated using an adjacent normal vertebral body as the reference area. RESULTS: Except for one patient, all other patients had increased uptake in the affected vertebra on bone scintigraphy [corrected]. In leukocyte scintigraphy, clearly decreased uptake was noted quantitatively in six of the seven patients who underwent SPECT. On the magnetic resonance and computed tomographic scans, the typical findings were destruction of intervertebral disks and compression deformities of vertebral bodies, but there was high interindividual variance of other findings, such as osteophytes, spondylolisthesis, increased vertebral fat content, and postoperative changes. These data show that nearly all patients with previously acute infective spondylitis have gross abnormal anatomic and functional imaging findings years after the acute disease, despite good clinical outcome [corrected]. CONCLUSIONS: The utility of bone and leukocyte scans in the diagnosis of reactivated spondylitis is limited, and incidentally observed abnormal imaging findings in asymptomatic patients with known previous spondylitis should be interpreted with caution. PMID- 11043717 TI - Fibrous dysplasia with barely increased uptake on bone scan: a case report. AB - Fibrous dysplasia, in general, appears as an area of markedly increased uptake on bone scintigraphy. Therefore, the possibility of fibrous dysplasia is likely to be excluded when the lesion shows no or slightly increased uptake. The authors report a case of incidentally detected fibrous dysplasia that appeared as slightly increased uptake on bone scintigraphy and was found to harbor a bone infarction along with typical fibrous dysplasia by pathologic examination of a specimen sampled by curettage. Barely increased bone uptake in fibrous dysplasia may be associated with decreased vascularity and osteoblast activity of the lesion as a result of concurrent bone infarction. The authors suggest that not every case of fibrous dysplasia appears as an area of intensely increased uptake on a bone scan. Clinicians should be cautious in interpreting bone scans of radiographically indicated fibrous dysplasia. PMID- 11043719 TI - Tc-99m MIBG imaging in a huge clinically silent pheochromocytoma with cystic degeneration and massive hemorrhage. AB - I-131 metaiodobenzylguanidine scintigraphy showed marked accumulation in the walls of a clinically silent, huge cystic adrenal mass with a prominent hemorrhage in a 48-year-old man. Although a careful reexamination of the histologic specimen finally lead to a diagnosis of pheochromocytoma, the appearances of this mass on computed tomography and magnetic resonance imaging were not specific for this neoplasm, and even pathologic analysis initially indicated, incorrectly, that this lesion was a hemorrhagic hemangioma. This case shows that I-131 metaiodobenzylguanidine scintigraphy is useful for correctly diagnosing an adrenal mass with prominent cystic or hemorrhagic degeneration. PMID- 11043718 TI - Imaging of malignant lymphomas with F-18 FDG coincidence detection positron emission tomography. AB - PURPOSE: The authors evaluated the utility of F-18 fluorodeoxyglucose (FDG) coincidence detection (CoDe) positron emission tomography (PET) for staging, post treatment evaluation, and follow-up assessment of patients with malignant lymphomas. MATERIALS AND METHODS: Fifty-eight patients with histologically proved malignant lymphomas (4 Hodgkin's disease, 54 non-Hodgkin's lymphoma) underwent CoDe PET using F-18 FDG. CoDe PET was performed using a dual-head gamma camera equipped with coincidence detection circuitry. Of the 87 CoDe PET studies, 26 were performed for staging, 38 for post-treatment evaluation, and 23 for follow up evaluation of recurrence. The entire trunk, from the cervical to the inguinal regions, or selected regions were scanned with the patient in the supine position. No attenuation correction was made and reconstruction was performed using filtered back-projection rather than iterative reconstruction. CoDe PET findings were compared with corresponding results of computed tomographic (CT) and magnetic resonance imaging (MRI), tissue biopsy, or clinical follow-up. RESULTS: For staging, 52 sites were positive on CoDe PET or CT-MRI. CoDe PET detected 49 sites (94%), and CT-MRI showed 47 sites (90%). CoDe PET detected five more lymphomatous lesions and missed three lesions. For post-treatment evaluation, CoDe PET showed a positive predictive value of 100% and a negative predictive value of 83%, but the validated cases numbered only 11. For follow-up for recurrence, CoDe PET had a negative predictive value of 90%, but frequent false-positive findings were noted in the head and neck region as a result of underlying inflammatory changes. CONCLUSIONS: For staging, FDG CoDe PET alone without attenuation correction is not sensitive enough to be used as an independent imaging method, especially for small abdominal lesions. However, it appears to be an accurate method for assessing residual disease and for patient follow-up. PMID- 11043720 TI - An unusual white blood cell scan in a child with inflammatory bowel disease: a case report. AB - Technetium-99m-labeled leukocyte (WBC) imaging is a valuable screening method for inflammatory bowel disease, especially in children, because of its high rate of sensitivity, low cost, and ease of preparation. A 14-year-old girl is described who had juvenile arthritis and iritis complicated by inflammatory bowel disease. She was examined for recurrent abdominal pain. A Tc-99m stannous colloid WBC scan was performed, and tracer accumulation was seen in the small bowel in the region of the distal ileum on the initial 1-hour image. Delayed imaging at 3 hours also revealed tracer accumulation in the cecum and ascending colon, which was not seen on the early image. A biopsy of the colon during endoscopy showed no evidence of active inflammation in the colon. The small bowel was not seen. Computed tomography revealed changes suggestive of inflammatory bowel disease in the distal ileum. The appearance on the WBC study was most likely a result of inflammatory bowel disease involving the distal ileum, with transit of luminal activity into the large bowel. PMID- 11043721 TI - Radionuclide imaging in patients with amputations of the lower leg: typical imaging patterns in five cases. AB - Amputation of the lower leg is not uncommon in elderly patients with chronic infections or vascular problems of the leg and foot, and most often it is performed below the knee or on the distal part of the lower leg (i.e., Syme's amputation). After operation, healing disorders with or without infection can occur, and usually structural imaging methods are performed for diagnosis. Radionuclide imaging using a combination of bone scans and infection scintigraphy can help to identify bone and soft tissue infection. Interpreting radionuclide scans is difficult, because imaging findings after amputation may depend not only on the level of resection and the disease investigated but also on the time that has elapsed since surgery. Typical imaging patterns of bone or infection scintigraphy of five patients are described PMID- 11043722 TI - Bone scintigraphy of elephantiasis neuromatosa in Von Recklinghausen's disease. PMID- 11043723 TI - Demonstration of non-small-cell lung carcinoma by indium-111 octreotide scintigraphy. PMID- 11043724 TI - Bone metastases in a patient with colon cancer depicted by Tc-99m carcinoembryonic antigen scintigraphy. PMID- 11043725 TI - Role of intraoperative skeletal scintigraphy in the localization of osteoblastomas. PMID- 11043726 TI - Unusual extravasation of urine diagnosed on bone scan. PMID- 11043727 TI - Renal scintigraphy with Tc-99m DTPA in pheochromocytoma: mistake or not? PMID- 11043728 TI - Curious radioactivity in the lower abdomen on bone scintigraphy: displacement of the urinary bladder by an incidentally diagnosed uterine myoma. PMID- 11043729 TI - Myocardial Tc-99m MDP uptake on a bone scan in senile systemic amyloidosis with cardiac involvement. PMID- 11043730 TI - Shunt imaging in a complex congenital cardiomyopathy. PMID- 11043731 TI - Isolated left hepatic lobe cholestasis demonstrated on cholescintigraphy. PMID- 11043732 TI - Central scar in hepatic focal nodular hyperplasia revealed by scintigraphy. PMID- 11043733 TI - False-positive imaging of Tc-99m pertechnetate Meckel's scintigraphy in a patient with hemorrhagic enteritis. PMID- 11043734 TI - Gastroesophageal reflux and myocardial imaging. PMID- 11043735 TI - Tc-99m red blood cell imaging of blue rubber bleb nevus syndrome. PMID- 11043736 TI - Detection of chronic recurrent lower extremity deep venous thrombosis on fluorine 18 fluorodeoxyglucose positron emission tomography. PMID- 11043737 TI - Preoperative and postoperative lymphoscintigraphy using Tc-99m sulfur colloid in the repair of a lymphatic leak in a patient with traumatic chylothorax. PMID- 11043738 TI - Imaging pelvic inflammatory disease with Tc-99m ciprofloxacin. PMID- 11043739 TI - Extraosseous accumulation of Tc-99m diphosphonate. PMID- 11043740 TI - TI-201 imaging in a malignant peripheral nerve sheath tumor. PMID- 11043741 TI - Visualization of the right atrial appendage during sestamibi scintigraphy. PMID- 11043742 TI - Cutaneous mantle cell lymphoma detected with Ga-67 Citrate. PMID- 11043743 TI - Circuit focus in early seizure after acute subcortical hemorrhage. PMID- 11043744 TI - Current readings in nuclear medicine. PMID- 11043745 TI - Simultaneous determination of ivabradine and its metabolites in human plasma by liquid chromatography--tandem mass spectrometry. AB - A rapid, selective, sensitive and reproducible liquid chromatographic method with tandem mass spectrometric detection has been developed and validated for the analysis of a new specific bradycardic agent, ivabradine (S 16257) and six potentially active metabolites in human plasma. Isolation of these compounds and of the internal standard was performed by an automated solid-phase extraction system using Oasis cartridges. Separation and detection of ivabradine and its metabolites were achieved using a C18 column and a MS-MS detector with a positive electrospray ionization source. Ivabradine and its metabolites gave a linear response ranging from 0.1 or 0.2 to 20 ng/ml and the limits of quantitation ranged from 0.1 to 0.2 ng/ml using a 0.5 ml plasma sample size. A complete validation demonstrated the method to be accurate, precise and specific for the simultaneous quantification of ivabradine and its metabolites in human plasma. The method was subsequently applied to the quantitative determination of ivabradine and its metabolites in human plasma samples from healthy volunteers participating in a clinical study to provide pharmacokinetic data. PMID- 11043746 TI - Assay of tramadol in urine by capillary electrophoresis using laser-induced native fluorescence detection. AB - Capillary electrophoresis (CE) with UV laser-induced native fluorescence detection was developed as a sensitive and selective assay for the direct determination of tramadol in human urine without extraction or preconcentration. The main problem in CE is the small inner diameter of the capillary which causes a low sensitivity with instruments equipped with a UV detector. Laser-induced native fluorescence with a frequency doubled argon ion laser at an excitation wavelength of 257 nm was used for the direct assay of tramadol in urine to enhance the limit of detection about 1,000-fold compared to UV absorption detection. The detection system consists of an imaging spectrograph and an intensified CCD camera, which views an illuminated 1.5 mm section of the capillary. This set-up is able to record the whole emission spectra of the analytes to achieve additionally wavelength-resolved electropherograms. In the concentration range of 20 ng/ml-5 microg/ml in human urine coefficients of correlation were better than 0.998. Within-day variation determined on four different concentrations showed accuracies ranging from 90.2 to 108.4%. The relative standard deviation (RSD) was determined to be less than 10%. Day-to-day variation presented accuracies ranging from 90.9 to 103.1% with an RSD less than 8%. PMID- 11043747 TI - High-performance liquid chromatographic analysis of muscular interstitial arginine and norepinephrine kinetics. A microdialysis study in rats. AB - Complex interactions between the L-arginine/nitric oxide synthase (NOS) pathway and the sympathetic nervous system have been reported. Methods capable of measuring L-arginine and norepinephrine (NE) have mainly been reported for plasma. We report the use of the microdialysis technique combined with high performance liquid chromatography (HPLC) for measurement of both L-arginine and NE within the same tissue microdialysis sample. The microdialysis probe consisted of linear flexible probes (membrane length: 10 mm, outside diameter: 290 microm, molecular weight cut-off 50 kDa). The method used for L-arginine measurement was HPLC with fluorescence detection, giving a within-run and a between-day coefficient of variation of 2.9 and 12.8%, respectively. The detection limit was 0.5 pM/20 microl injected for L-/D-arginine. The method used for NE measurement was HPLC with electrochemical detection. The coefficients of variation were 4% for within-assay precision and 7.5% for between-assay precision. The detection limit for NE was 1 fmol/20 microl injected. The microdialysis technique coupled with HPLC system was validated in vivo to measure muscular interstitial concentrations of both arginine and NE under baseline conditions and after intravenous infusion of 500 mg/kg of L-arginine or D-arginine. In conclusion, the microdialysis technique coupled to HPLC allows the simultaneous measurements of both L-arginine and NE within the same tissue microenvironment and will enable the study of the complex interactions between the L-arginine/NO pathway and sympathetic nervous system within the interstitial space of different organs. PMID- 11043748 TI - Liquid chromatography--tandem mass spectrometry analysis of cocaine and its metabolites from blood, amniotic fluid, placental and fetal tissues: study of the metabolism and distribution of cocaine in pregnant rats. AB - The ability to simultaneously quantitate cocaine and its 12 metabolites from pregnant rat blood, amniotic fluid, placental and fetal tissue homogenates aids in elucidating the metabolism and distribution of cocaine. An efficient extraction method was developed to simultaneously recover these 13 components using underivatized silica solid-phase extraction (SPE) cartridges. The overall recoveries for cocaine and its metabolites were studied from pregnant rat blood (47-100%), amniotic fluid (61-100%), placental homogenate (31-83%), and fetal homogenate (39-87%). Extraction of the samples using silica is not classical SPE, but rather allows for the concentration of the sample into a small volume prior to injection and the removal of the proteins due to their strong interaction with the active silica surface. A positive ion mode electrospray ionization liquid chromatography-tandem mass spectrometry (LC-MS-MS) method was used and validated to simultaneously quantitate cocaine and 12 metabolites from these four biological matrices. A gradient elution method with a Zorbax XDB C8 reversed phase column was used to separate the components. Multiple reaction monitoring (MRM) of a product ion arising from the corresponding precursor ion was used in order to enhance the selectivity and sensitivity of the method. Low background noise was observed from the complex biological matrices due to efficient SPE and the selectivity of the MRM mode. Linear calibration curves were generated from 0.01 to 2.50 ppm. The method also showed high intra-day (n =3) and inter-day (n=9) precision (% RSD) and accuracy (% error) for all components. The limits of detection (LODs) for the method ranged from 0.15 to 10 ppb. The LODs of cocaine and its major metabolites were less than 1 ppb from all four biological matrices. This method was applied to the study of the metabolism and distribution of cocaine in pregnant rats following intravenous infusion to a steady state plasma drug concentration. The following results were observed in the pregnant rat study: (1) the observations correlated strongly with the previous literature data on cocaine metabolism and distribution, (2) cocaine and norcocaine accumulated in the placenta, (3) arylhydroxylation of cocaine was a major metabolic pathway, (4) para-arylhydroxylation of cocaine was favored over meta-arylhydroxylation in rats and (5) accumulation of cocaine and its major metabolites was observed in the amniotic fluid. PMID- 11043749 TI - High-performance liquid chromatography with on-line post-column immunoreaction detection of digoxin and its metabolites based on fluorescence energy transfer in the far-red spectral region. AB - The combination of immunoassays with separation techniques such as chromatography can result in enhanced selectivity and sensitivity. This paper describes an on line chromatography with immunochemical post-column fluorescence energy transfer detection for digoxin and its metabolites. R-phycoerythrin (PE) was used as the donor and an indodicarbocyanine dye (Cy5) as the acceptor label. These labels allow the detection in the far-red spectral region, which is more selective for biological samples. Hence, digoxin was labeled with PE using the activated digoxigenin-NHS-ester and monoclonal anti-digoxin antibody was labeled with Cy5. Digoxin and its metabolites was injected into the HPLC system followed by post column injection of R-phycoerythrin labeled digoxin and by Cy5 labeled anti digoxin antibody. Incubation time was provided using an open tubular reactor coil at room temperature. The detection was performed by measurement of the sensitized emission of Cy5 at 670 nm due to fluorescence energy transfer from PE labeled with digoxin. The system was optimized with regard to the concentrations of the used post-column reagents as well as incubation time and temperature. The dynamic range of digoxin spiked in 0.01 M phosphate buffer (pH 7.4) was 0.05 to 10 ng/ml with a correlation coefficient of 0.989. The limit of detection was 33 pg/ml. The precision of two controls, 0.4 and 4 ng/ml, was found to be 2.2 and 8.7% RSD, respectively, accuracy was 10.7 and 20.3% (n=6 in each case). PMID- 11043750 TI - High-performance liquid chromatographic determination of cocaine and its metabolites in serum microsamples with fluorimetric detection and its application to pharmacokinetics in rats. AB - A sensitive, selective and simple HPLC method with fluorimetric detection is described for quantitating cocaine and its three metabolites in rat serum microsamples (50 microl). Chromatographic separation is achieved on a Hypersil BDS C18 column (100X2.1 mm, 5 microm) with an isocratic mobile phase consisting of methanol-acetonitrile-25.8 mM sodium acetate buffer, pH 2.6, containing 1.0 x 10(-4) M tetrabutylammonium phosphate (14:10:76, v/v/v). The detection limit (0.5 ng/ml) for all the compounds, using direct fluorometric detection operated at excitation and emission wavelengths of 230 and 315 nm, respectively, was approximately five-times lower than that of using a UV detector operated at 235 nm. The effects of ratio of 2-propanol to chloroform in extraction solvents on the recovery and precision for cocaine and its metabolites were systematically examined. The method was used to study the pharmacokinetics of cocaine after administration of intravenous 2 mg/kg and oral 20 mg/kg doses. PMID- 11043751 TI - Enantioselective analysis of levetiracetam and its enantiomer R-alpha-ethyl-2-oxo pyrrolidine acetamide using gas chromatography and ion trap mass spectrometric detection. AB - A gas chromatographic-mass spectrometric method was developed for the enantioselective analysis of levetiracetam and its enantiomer (R)-alpha-ethyl-2 oxo-pyrrolidine acetamide in dog plasma and urine. A solid-phase extraction procedure was followed by gas chromatographic separation of the enantiomers on a chiral cyclodextrin capillary column and detection using ion trap mass spectrometry. The fragmentation pattern of the enantiomers was further investigated using tandem mass spectrometry. For quantitative analysis three single ions were selected from the enantiomers, enabling selected ion monitoring in detection. The calibration curves were linear from 1 microM to 2 mM for plasma samples and from 0.5 mM to 38 mM for urine samples. In plasma and urine samples the inter-day precision, expressed as relative standard deviation was around 10% in all concentrations. Selected ion monitoring mass spectrometry is suitable for quantitative analysis of a wide concentration range of levetiracetam and its enantiomer in biological samples. The method was successfully applied to a pharmacokinetic study of levetiracetam and (R)-alpha-ethyl-2-oxo-pyrrolidine acetamide in a dog. PMID- 11043752 TI - Determination of perfluorodecalin and perfluoro-N-methylcyclohexylpiperidine in rat blood by gas chromatography-mass spectrometry. AB - A gas chromatography-mass spectrometry method (SIM mode) was developed for the determination of perfluorodecalin (cis and trans isomers, 50% each) (FDC), and perfluoromethylcyclohexylpiperidine (3 isomers) (FMCP) in rat blood. The chromatographic separation was performed by injection in the split mode using a CP-select 624 CB capillary column. Analysis was performed by electronic impact ionization. The ions m/z 293 and m/z 181 were selected to quantify FDC and FMCP due to their abundance and to their specificity, respectively. The ion m/z 295 was selected to monitor internal standard. Before extraction, blood samples were stored at -30 degrees C for at least 24 h in order to break the emulsion. The sample preparation procedure involved sample clean-up by liquid-liquid extraction. The bis(F-butyl)ethene was used as the internal standard. For each perfluorochemical compound multiple peaks were observed. The observed retention times were 1.78 and 1.87 min for FDC, and 2.28, 2.34, 2.48 and 2.56 min for FMCP. For each compound, two calibration curves were used; assays showed good linearity in the range 0.0195-0.78 and 0.78-7.8 mg/ml for FDC, and 0.00975-0.39 and 0.39 3.9 mg/ml for FMCP. Recoveries were 90 and 82% for the two compounds, respectively with a coefficient of variation <8%. Precision ranged from 0.07 to 15.6%, and accuracy was between 89.5 and 111.4%. The limits of quantification were 13 and 9 microg/ml for FDC and FMCP, respectively. This method has been used to determine the pharmacokinetic profile of these two perfluorochemical compounds in blood following administration of 1.3 g of FDC and 0.65 g of FMCP per kg body weight, in emulsion form, in rat. PMID- 11043753 TI - Simple and selective determination of the cyclophosphamide metabolite phosphoramide mustard in human plasma using high-performance liquid chromatography. AB - A simple and selective assay for the determination of the alkylating cyclophosphamide metabolite phosphoramide mustard (PM) in plasma was developed and validated. PM was determined after derivatisation by high-performance liquid chromatography (HPLC) with ultraviolet detection at 276 nm. Sample pre-treatment consisted of derivatisation of PM with diethyldithiocarbamate (DDTC) at 70 degrees C for 10 min, followed by extraction with acetonitrile in the presence of 0.7 M sodium chloride. Phase separation occurred due to the high salt content of the aqueous phase. The HPLC system consisted of a C8 column with acetonitrile 0.025 M potassium phosphate buffer, pH 8.0, (32:68, v/v) as the mobile phase. The entire sample handling procedure, from collection at the clinical ward until analysis in the laboratory, was optimised and validated. Calibration curves were linear from 50 to 10,000 ng/ml. The lower limit of quantification and the limit of detection (using a signal-to-noise ratio of 3) were 50 and 40 ng/ml, respectively, using 500 microl of plasma. Within-day and between-day precisions were below 11% over the entire concentration range and the accuracies were between 100 and 106%. PM was found to be stable at -30 degrees C for at least 10 weeks both in plasma and as a DDTC-derivative in a dry sample. A pharmacokinetic pilot study in two patients receiving 1,000 mg/m2 CP in a 1-h infusion demonstrated the applicability of the assay. PMID- 11043754 TI - High-performance liquid chromatography of the neuroactive steroids alphaxalone and pregnanolone in plasma using dansyl hydrazine as fluorescent label: application to a pharmacokinetic-pharmacodynamic study in rats. AB - This report describes a rapid and sensitive analytical method for the quantification of the neuroactive steroids alphaxalone and pregnanolone in rat plasma using derivatization with dansyl hydrazine as fluorescent label. The method involves protein precipitation, alkaline derivatization and extraction of the compounds and internal standard pregnenolone with dichloromethane, followed by isocratic reversed-phase high-performance liquid chromatography on a 3-microm Microsphere C18 column with fluorescence detection at wavelengths 332 nm and 516 nm for excitation and emission, respectively. The mobile phase consists of a mixture of 25 mM acetate buffer (pH 3.7)-acetonitrile (45:55, v/v for alphaxalone and 40:60, v/v for pregnanolone) with a flow-rate of 1 ml/min. The total run time was approximately 35 min. In the concentration range of 0.010-10 microg ml(-1), the intra- and inter-assay coefficients of variation were less than 17% for both methods. In 50 microl plasma samples the corresponding limits of detection were 10 ng ml(-1) (signal-to-noise ratio=3). The utility of the analytical method was established by analyzing plasma samples from rats, which had received an intravenous administration of 5 mg kg(-1) alphaxalone or pregnanolone. Values for clearance, volume of distribution at steady state and terminal half life were 71.9 ml min(-1) kg(-1), 814 mg kg(-1) and 13.5 min for alphaxalone and 69.2 ml min(-1) kg(-1), 1,638 ml kg(-1) and 27.8 min for pregnanolone, respectively. Due to its simplicity and sensitivity this method can be used on a routine basis for pharmacokinetic analysis of neuroactive steroids. PMID- 11043755 TI - High resolution of oligosaccharide mixtures by ultrahigh voltage micellar electrokinetic capillary chromatography. AB - Oligosaccharide mixtures released from ribonuclease B and human IgG have been separated using micellar electrokinetic capillary chromatography operated at 100 kV. The resolution of these closely related analytes at this high voltage was found to be superior to that obtained at 20 kV, a voltage which is ordinarily used in most capillary electrophoresis separations. PMID- 11043756 TI - Development of a high-performance liquid chromatographic-mass spectrometric assay for the specific and sensitive quantification of Ro 64-0802, an anti-influenza drug, and its pro-drug, oseltamivir, in human and animal plasma and urine. AB - Oseltamivir phosphate (Ro 64-0796/002) is a pro-drug of the anti-influenza neuraminidase inhibitor, Ro 64-0802, and as Tamiflu, has been developed for the treatment of both A and B strains of the disease. This paper describes an HPLC-MS MS assay for both compounds in plasma and urine which fulfils all of the criteria for a good analytical method. It is sensitive with limits of quantification of 1 and 10 ng/ml for the pro-drug and active neuraminidase inhibitor, respectively. It is both accurate and precise with typical coefficients of variation from some 5,000 quality control samples of approximately +/-3 and +/-6%, respectively. Extensive stability studies have demonstrated the absence of significant problems associated with the decomposition of either compound, although ex vivo hydrolysis of Ro 64-0796 to Ro 64-0802 in rodent plasma has to be prevented by the use of the esterase inhibitor, dichlorvos. PMID- 11043757 TI - Chiral separation of amino acids in biological fluids by micellar electrokinetic chromatography with laser-induced fluorescence detection. AB - A method is presented for the chiral analysis of amino acids in biological fluids using micellar electrokinetic chromatography (MEKC) and laser-induced fluorescence (LIF). The amino acids are derivatized with the chiral reagent (+/-) 1-(9-anthryl)-2-propyl chloroformate (APOC) and separated using a mixed micellar separation system. No tedious pre-purification of samples is required. The excellent separation efficiency and good detection capabilities of the MEKC-LIF system are exemplified in the analysis of urine and cerebrospinal fluid. This is the first time MEKC has been reported for chiral analysis of amino acids in biological fluids. The amino acids D-alanine, D-glutamine, and D-aspartic acid have been observed in cerebrospinal fluid, and D-alanine and D-glutamic acid in urine. To the best of our knowledge no measurements of either D-alanine in cerebrospinal fluid or D-glutamic acid in urine have been presented in the literature before. PMID- 11043758 TI - High-temperature solid-phase microextraction procedure for the detection of drugs by gas chromatography-mass spectrometry. AB - High-temperature headspace solid-phase microextraction (SPME) with simultaneous ("in situ") derivatisation (acetylation or silylation) is a new sample preparation technique for the screening of illicit drugs in urine and for the confirmation analysis in serum by GC-MS. After extraction of urine with a small portion of an organic solvent mixture (e.g., 2 ml of hexane-ethyl acetate) at pH 9, the organic layer is separated and evaporated to dryness in a small headspace vial. A SPME-fiber (e.g., polyacrylate) doped with acetic anhydride-pyridine (for acetylation) is exposed to the vapour phase for 10 min at 200 degrees C in a blockheater. The SPME fiber is then injected into the GC-MS for thermal desorption and analysis. After addition of perchloric acid and extraction with n hexane to remove lipids, the serum can be analysed after adjusting to pH 9 as described for urine. Very clean extracts are obtained. The various drugs investigated could be detected and identified in urine by the total ion current technique at the following concentrations: amphetamines (200 microg/l), barbiturates (500 microg/l), benzodiazepines (100 microg/l), benzoylecgonine (150 microg/l), methadone (100 microg/l) and opiates (200 microg/l). In serum all drugs could be detected by the selected ion monitoring technique within their therapeutic range. As compared to liquid-liquid extraction only small amounts of organic solvent are needed and larger amounts of the pertinent analytes could be transferred to the GC column. In contrast to solid-phase extraction (SPE), the SPME-fiber is reusable several times (as there is no contamination by endogenous compounds). The method is time-saving and can be mechanised by the use of a dedicated autosampler. PMID- 11043759 TI - Liquid chromatographic-mass spectrometric determination of itraconazole and its major metabolite, hydroxyitraconazole, in dog plasma. AB - A fast, reliable and sensitive liquid chromatography-mass spectrometry (LC-MS) assay for the determination of itraconazole and hydroxyitraconazole in dog plasma has been developed. The analysis involves a simple liquid-liquid extraction followed by LC-MS analysis using electrospray ionization in the positive mode. Total separation of itraconazole, hydroxyitraconazole and the internal standard, miconazole, was achieved on a C18 column in 3.5 min using an isocratic mixture of acetonitrile and 10 mM ammonium acetate. The response was linear over four-orders of magnitude, allowing reliable quantification of each species. This paper describes the development of the method and its validation. PMID- 11043760 TI - High-performance liquid chromatography method for the determination and quantitation of arabinosylguanine triphosphate and fludarabine triphosphate in human cells. AB - A gradient anion-exchange high-performance liquid chromatographic assay was developed for the simultaneous determination and quantitation of the cytotoxic triphosphates of arabinosylguanine (ara-GTP) and fludarabine (F-ara-ATP). The method was validated with respect to selectivity, recovery, linearity, precision, and accuracy using authentic standards. To test this assay in a more complex biological matrix, perchloric acid extracts of circulating human leukemia cells spiked with known concentrations of ara-GTP and F-ara-ATP were examined. Finally, to assess the clinical utility of our method, perchloric acid extracts of circulating human leukemia cells isolated from patients treated with fludarabine and nelarabine were analyzed. The range of quantitation was 0.0125-10 nmol for the ara- and native NTPs in cellular extracts. This assay should be helpful in establishing the mechanistic rationales for drug scheduling and combinations of nelarabine and fludarabine, and for correlating the therapeutic efficacy and levels of the cytotoxic triphosphates in target cells. PMID- 11043761 TI - Rapid and simultaneous high-performance liquid chromatography assay of polyamines and monoacetylpolyamines in biological specimens. AB - A rapid, resolutive and reproducible reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed for polyamines and acetylpolyamines by adopting pre-column derivatization with benzoyl chloride. In a single run lasting less than 15 min ten polyamines were separated as well as traces of benzoic acid, methylbenzoate and benzoic anhydride. These contaminants, produced during the derivatization reaction, were almost all eliminated by washing steps envisaged in the same procedure. This simple and sensitive method can be applied to routine determination of polyamines in biological samples. A fine application of this procedure to the determination of endogenous content of polyamines in chick embryo retina was reported. PMID- 11043762 TI - Improved high-performance liquid chromatographic analysis of terazosin in human plasma. AB - A simple, sensitive and reproducible high-performance liquid chromatography (HPLC) method was developed for the determination of terazosin in human plasma. The method involves a one-step single solvent extraction procedure using dichloromethane with a 0.25 ml plasma sample. Recovery values were all greater than 90% over the concentration range 0.25-100 ng/ml. Terazosin was found to adsorb to glass or plastic tubes, but this could be circumvented by using disposable plastic tubes. Also, rinsing the injector port with methanol after each injection helped to prevent any carry-over effect. The internal standard, prazosin, did not exhibit this problem. The method has a quantification limit of 0.25 ng/ml. The within- and between-day coefficient of variation and accuracy values were all less than 7% over the concentration range 0.25-100 ng/ml and hence the method is suitable for use in pharmacokinetic studies of terazosin. PMID- 11043763 TI - B-cell activation 2000. PMID- 11043764 TI - An unsolved problem of the clonal selection theory and the model of an oligomeric B-cell antigen receptor. AB - The B cell antigen receptor (BCR) plays a central role in the development, survival and activation of B lymphocytes. As the pre-BCR, it controls allelic exclusion of heavy chains and the expansion of pre-B cells. As the BCR, it controls the positive and negative selection of immature B cells as well as the survival and activation of mature B cells. Recent studies of receptors have shown that it is the ligand that brings them into the conformation necessary for signaling. How the multiple and structurally diverse antigens could fulfill this task for the BCR is unknown, and we regard this as an unsolved problem of Burnet's clonal selection theory This question and our recent biochemical studies lead us to propose a new model for the BCR, according to which the BCR exists as a precise oligomeric complex on the B cell surface. In this form, it can signal positive selection and survival of B cells. Binding to self- or foreign antigen results in a distortion of the oligomeric complex that gives the signal for negative selection of immature and activation of mature B cells. PMID- 11043765 TI - Regulation of the phospholipase C-gamma2 pathway in B cells. AB - In B lymphocytes, a signaling complex that contributes to cell fate decisions is the B-cell antigen receptor (BCR), with different extents of receptor engagement leading to such outcomes as cell death, survival, or proliferation. Here, based upon the available genetic and biochemical data of the BCR signal components, we discuss several mechanisms by which BCR signals are propagated and modified, with specific emphasis on the phospholipase C (PLC)-gamma2-calcium pathway Gene targeting experiments in DT40 chicken B cells highlighted the importance of the intracellular protein tyrosine kinases Syk and Btk in PLC-gamma2 activation. Until recently, the molecular mechanism underlying the double requirement for Syk and Btk in PLC-gamma2 activation remained unclear, but new data suggest that an adapter molecule, B-cell linker protein (alternatively named SLP-65 or BASH), phosphorylated by Syk, provides docking sites for Btk SH2 domain as well as PLC gamma2 SH2 domains, thus bringing Btk into close proximity with PLC-gamma2. The activated Btk then phosphorylates PLC-gamma2, leading to its activation. The activated PLC-gamma2 converts phosphatidylinositol 4,5-bisphosphate into the second messenger inositol 1,4,5-trisphosphate (IP3), which in turn binds to IP3 receptors located on the endoplasmic reticulum (ER). Binding of IP3 to the IP3 receptors is essential for triggering a calcium release from the ER and subsequent entry of extracellular calcium. Balancing these activation signals in the PLC-gamma2-calcium pathway are the inhibitory receptors expressed on B cells, FcyRII and paired immunoglobin-like receptor (PIR)-B. Although both FcyRII and PIR-B inhibits the BCR-mediated [Ca2+]i increase, the inhibitory mechanisms of these receptors are distinct. The FcyRII-mediated inhibitory signal is dependent on lipid phosphatase SHIP, whereas the PIR-B requires redundant functions of protein phosphatases SHP-1 and SHP-2. Thus, PIR-B and FcgammaRII inhibit calcium signals by utilizing two distinct signaling molecules, thereby contributing to setting threshold levels for activation signals as well as terminating activation responses. PMID- 11043766 TI - Regulation of B-cell activation and differentiation by the phosphatidylinositol 3 kinase and phospholipase Cgamma pathway. AB - Signal transduction through the B-cell antigen receptor (BCR) determines the fate of B lymphocytes during their development and during immune responses. A multitude of signal transduction events are known to be activated by ligation of the BCR; however, the critical parameters determining the biological outcome of the signal transduction cascade are only just beginning to be understood. Two enzymes which act on plasma membrane phospholipids, phosphatidylinositol 3-kinase (PI3K) and phospholipase Cgamma (PLCgamma), have been implicated as critical mediators of B-cell activation and differentiation signals. Activation of these ubiquitous enzymes is regulated by B-lymphocyte-specific signal transduction proteins, such as CD 19 and B-cell linker protein. These enzymes function by generating both membrane-anchored and soluble second messenger molecules which regulate the activity of downstream signal transduction proteins. Active PI3K produces phosphatidylinositol-3,4-bisphosphate (PI(3,4)P2) and phosphatidylinositol-3,4-trisphosphate (PI(3,4,5)P3) which can bind to signaling proteins such as Btk or Akt via their pleckstrin homology domains, resulting in their membrane recruitment and activation. The lipid phosphatases SHIP and PTEN negatively regulate production of PI(3,4)P2 and PI(3,4,S)P3 and therefore function to put a "brake" on the PI3K pathway. Active PLCgamma produces inositol 1,4,5-trisphosphate, which regulates Ca2+ mobilization, and diacylglycerol, which binds to a subset of protein kinase C enzymes leading to their membrane localization and activation. Recent evidence has indicated that PLCgamma activation is partially dependent on the PI(3,4,5)P3 production by activated PI3K. Since PI3K and PLCgamma also share common downstream targets such as the NF AT and NF-kappaB transcription factors, it is becoming clear that these two pathways are interconnected at several levels. Studies of mice deficient in components of the PI3K and PLCgamma pathways demonstrate that these pathways play critical roles in both pre-BCR and BCR-dependent selection events during B-cell differentiation. Taken together, the present data clearly indicate that PI3K and PLCgamma play critical and indispensable roles in the signal transduction cascades leading to multiple biological responses downstream of the BCR. PMID- 11043767 TI - Targets of B-cell antigen receptor signaling: the phosphatidylinositol 3 kinase/Akt/glycogen synthase kinase-3 signaling pathway and the Rap1 GTPase. AB - In this review, we discuss the role of phosphatidylinositol 3-kinase (PI3K) and Rap 1 in B-cell receptor (BCR) signaling. PI3K produces lipids that recruit pleckstrin homology domain-containing proteins to the plasma membrane. Akt is a kinase that the BCR activates in this manner. Akt phosphorylates several transcription factors as well as proteins that regulate apoptosis and protein synthesis. Akt also regulates glycogen synthase kinase-3, a kinase whose substrates include the nuclear factor of activated T cells (NF-AT)cl and beta catenin transcriptional activators. In addition to Akt, PI3K-derived lipids also regulate the activity and localization of other targets of BCR signaling. Thus, a key event in BCR signaling is the recruitment of PI3K to the plasma membrane where its substrates are located. This is mediated by binding of the Src homology (SH) 2 domains in PI3K to phosphotyrosine-containing sequences on membrane associated docking proteins. The docking proteins that the BCR uses to recruit PI3K include CD19, Cbl, Gab1, and perhaps Gab2. We have shown that Gab1 colocalizes PI3K with SH2 domain-containing inositol phosphatase (SHIP) and SHP2, two enzymes that regulate PI3K-dependent signaling. In contrast to PI3K, little is known about the Rap1 GTPase. We showed that the BCR activates Rap1 via phospholipase C-dependent production of diacylglycerol. Since Rap1 is thought to regulate cell adhesion and cell polarity, it may be involved in B-cell migration. PMID- 11043768 TI - Bilevel control of B-cell activation by the inositol 5-phosphatase SHIP. AB - The balanced interplay between positive and negative signals pathways emanating from surface receptors has emerged as a common paradigm for regulation of cell function and the immune response. Here, we will review the recent progress in analysis of signaling pathways initiated upon antigen receptor (BCR) aggregation, and co-aggregation with the inhibitory IgG receptor FcgammaRIIB. Particular attention is paid to the function of the inositol 5-phosphatase SHIP and its effector p62i(Dok), a RasGAP adapter protein. SHIP and Dok function in FcgammaRIIB-mediated inhibition as well as in feedback regulation of signals generated through the BCR. These inhibitory molecules may play critical roles in the prevention of immune system hyperactivity and resulting autoimmunity. PMID- 11043769 TI - CD72, a negative regulator of B-cell responsiveness. AB - The ability of lymphocytes to respond to antigenic or mitogenic stimulation is regulated not only by specific receptor proteins, but also by both positive and negative regulatory proteins that set or fine-tune the threshold for responsiveness. CD72 is one such regulatory protein on B lymphocytes. It is a member of the C-type lectin superfamily and is expressed on the surface of B cells from the pro-B through the mature B-cell stage. Studies with anti-CD72 antibodies have suggested a positive regulatory role for CD72 in B-cell activation. However, the cytoplasmic tail of CD72 contains two potential immunoreceptor tyrosine-based inhibitory motifs, one of which has been shown to recruit the tyrosine phosphatase SHP- 1. These features suggest a negative regulatory role for CD72. We have generated CD72-deficient mice to elucidate the physiological role of CD72 in B-lymphocyte development and activation. Our analyses of these mice and their B-cell compartment demonstrate that CD72 is a nonredundant regulator of B-cell development and a negative regulator of B-cell responsiveness. PMID- 11043770 TI - Immunobiology of the immature B cell: plasticity in the B-cell antigen receptor induced response fine tunes negative selection. AB - The immature and transitional immature B-cell stages define an important window in B-cell development, as it is at this point that cells committed to the B-cell lineage first express the clonotypic B-cell antigen receptor (BCR) and cells expressing self-reactive specificities may be identified and eliminated. The intrinsic susceptibility of the immature B cell to negative selection following BCR engagement distinguishes these cells functionally from mature-stage B cells in which BCR cross-linking leads to activation. Our laboratory has been interested in determining the molecular events responsible for the distinct and disparate responses of immature and mature B cells to antigen receptor signaling in order to understand the molecular basis of negative selection of developing B cells. These studies have indicated that developmentally regulated mechanisms, intrinsic to the B cell, regulate the differential responsiveness of the immature and mature stage B cell to antigen. However, the "hard-wired" BCR-induced apoptotic response of the immature B cell can be modified by the microenvironmental context in which the antigen is encountered. This plasticity fine tunes the BCR-induced response of the immature B cell by regulating the mechanism of negative selection and, under defined circumstances, allowing for recruitment into an immune response. PMID- 11043771 TI - B-cell receptor and Fas-mediated signals for life and death. AB - A series of B-cell lymphoma lines with an immature phenotype has been used as a model system to study molecular events associated with receptor ligation induced death. B-cell receptor (BCR) cross-linking with antibodies to membrane IgM (but not with anti IgD) induces c-Myc downregulation via nuclear factor kappaB inactivation and p27(Kip1) accumulation in these B lymphomas. Anti-mu-treated cells then undergo G1 arrest and die by apoptosis independent of Fas. Steroids and retinoids similarly downregulate c-Myc and induce apoptosis in these B cells and synergize with anti-mu. Rescue from apoptosis induced by anti-mu or steroids occurs with T-cell signals, like CD40L, or a broad-range caspase inhibitor, but only CD40L prevents the loss of c-Myc, p27 accumulation and growth arrest. Both IgM and IgD signaling lead to modulation of phosphatidylinositol 3-kinase (PI3K) signals, including the activation of p70(S6K), but this pathway recovers under anti-IgD treatment. Blockade of the PI3K pathway augments anti-mu-induced death and converts anti-delta to an apoptotic signal. Resistance to Fas-mediated death may be an important factor in B-cell transformation in vivo. Many of our panel of lymphomas are insensitive to Fas-mediated death signals, although all can form a death-inducing signaling complex (DISC). Additional studies suggest that some lymphomas can be blocked at the DISC complex by anti-apoptotic proteins, whereas others are inhibited downstream of caspase 8 activation. Anti-Ig treatment of a Fas-sensitive line, A20.2J, activated a number of genes whose products may block apoptosis proximally (like FLICE-inhibitory protein (FLIP1)) or at late points, such as bcl-2-family members. Our data suggest that B lymphomas develop multiple pathways of resistance to Fas-mediated signals during lymphomagenesis, in part via signaling through the BCR. PMID- 11043772 TI - Receptor-specific regulation of B-cell susceptibility to Fas-mediated apoptosis and a novel Fas apoptosis inhibitory molecule. AB - The susceptibility of primary B cells to Fas (APO-1, CD95)-mediated apoptosis is modulated by signals derived from additional surface receptors: CD40 engagement produces upregulation of Fas expression and marked sensitivity to Fas-induced cell death, whereas antigen receptor engagement, or interleukin-4 receptor (IL 4R) engagement, inhibits Fas killing and thereby produces Fas resistance, even in otherwise susceptible, CD40-stimulated targets. Surface immunoglobulin (sIg) and IL-4R utilize distinct signaling pathways to produce Fas resistance that rely on protein kinase C and signal transducer and activator of transcription 6, respectively sIg signaling for inducible Fas resistance requires nuclear factor kappaB and depends on new macromolecular synthesis. Proximate mediators for Fas resistance include the known anti-apoptotic gene products Bcl-xL and FLIP (but not Btk), and a novel anti-apoptotic gene that encodes Fas apoptosis inhibitory molecule (FAIM). FAIM was identified by differential display and was cloned as two alternatively spliced forms: FAIM-S is broadly expressed, whereas faim-L expression is tissue specific. faim is highly evolutionarily conserved, suggesting an important function throughout phylogeny. Inducible resistance to Fas-mediated apoptosis is speculated to protect antigen-specific B cells during potentially dangerous interactions with FasL-bearing T cells; the elevated sIg signaling threshold for inducible Fas resistance in autoreactive, tolerant B cells would insure against autoimmunity. However, aberrant acquisition of Fas resistance may allow autoreactive B cells to escape Fas deletion and malignant lymphocytes to thwart antitumor immunity. PMID- 11043773 TI - Rel/NF-kappaB transcription factors: key mediators of B-cell activation. AB - The Rel/nuclear factor (NF)-kappaB family of transcription factors have been implicated in the regulation of a wide variety of genes, in particular those encoding proteins crucial to the function of the immune system. Through the use of mutant mice that lack one or more of these proteins, we have begun to examine the individual and combined roles of Rel, RelA and NF-kappaB1 in B-cell development and function. Here we outline and discuss how these transcription factors operate as differentiation stage-specific regulators of B-cell development, survival, division and immunoglobulin expression, emphasizing those Rel/NF-kappaB-regulated genes that mediate these functions. PMID- 11043774 TI - B-cell antigen receptor competence regulates B-lymphocyte selection and survival. AB - Experimental evidence contradicts the simplistic view that during development all B cells expressing non autoreactive antigen receptors on the cell surface are selected into the mature B-cell pool. While allelic exclusion, clonal selection and affinity maturation continue to define the mainstream notions of B-cell development and selection, new evidence is redefining our understanding of these processes. Receptor editing replaces functional B-cell receptors by secondary immunoglobulin gene rearrangements, a process that can play roles in both immune tolerance and immune response. In addition, editing can rescue cells that would otherwise fail positive selection. We focus here on our studies indicating that the functional competence of the B-cell antigen receptor complex plays a central role in the fate of developing B cells and their antigen receptor genes. PMID- 11043775 TI - B-cell activation by T-cell-independent type 2 antigens as an integral part of the humoral immune response to pathogenic microorganisms. AB - Antigens that are expressed on the surface of pathogens in an organized, highly repetitive form can activate specific B cells by cross-linking of antigen receptors in a multivalent fashion. B cells respond to these multivalent antigens in the absence of MHC class II-restricted T-cell help by a mechanism that depends on the expression of a functional Bruton's tyrosine kinase (Btk). Accordingly, this class of immunogens has been designated T-cell-independent type 2 (TI-2) antigens. The unique properties of the B-cell response to TI-2 antigens are critically dependent on the formation of a small number of antigen receptor clusters, each of which contains approximately 10 to 20 antigen-bound membrane Ig (mIg) molecules. These clusters induce local membrane association of multiple activated Btk molecules, which results in long-term mobilization of intracellular ionized calcium. Such persistent calcium fluxes efficiently recruit transcription factors and thereby induce T-cell-independent B-cell activation and proliferation. While this first signal of multivalent mIg cross-linking can induce B-cell proliferation, we propose that a second signal is required for a TI 2 Ig secretory response. We have found that engagement of members of the Toll like receptor (TLR) family could provide second signals that selectively induce Ig secretion in B cells that were activated by multivalent, but not by bivalent, antigen receptor engagement. This finding demonstrates a general mechanism by which TLRs recognize molecular motifs on the surface of pathogens and provide the TI-2-activated B cell with a second signal. In addition, TLR-dependent recognition of these non-self motifs by cells of the innate immune system can induce these cells to provide alternative and/or additional second signals in the TI-2 response. The complement system provides another link between the B cell and the innate immune system, and facilitates the mIg signal transduction by recruitment of CD21 in the immune response. Thus, the TI-2 response provides the host with a combination of "the best of both worlds": the recruitment of the fine specificity of the adaptive immune response and the utilization of both the speed of the innate immune system and the wealth of cytokines produced by its member cells upon stimulation by pathogenic organisms or their products. By combining these two pathways, the TI-2 response enables the host to rapidly produce antigen specific Ig effector molecules that can be secreted at a sufficient rate to keep up with the rapid multiplication of invading infectious microorganisms, and will also prevent the intracellular spreading of a significant part of this population. PMID- 11043776 TI - A two-phase model of B-cell activation. AB - The current paradigm of lymphocyte activation, the two-signal model, has developed from the premise that recognition of antigen alone is insufficient to stimulate naive B cells, as this could potentially induce autoreactive responses, and that cognate T-B interaction is necessary to induce a full B-cell response. Recent evidence suggests, however, that T-cell-independent B-cell activation is part of the humoral immune response to pathogens, and therefore that antigen alone, or antigen plus signals provided by cells other than T cells, can provide all the necessary signals to induce a B-cell response. Furthermore, the presence of secreted IgM produced either as natural antibodies by CD5+ B-1 cells or as antigen-induced IgM by conventional (B-2) cells was shown to affect the kinetics and magnitude of the IgG response significantly. These data and the observed rapid kinetics of in vivo humoral responses seem at odds with a model that predicts that full B-cell activation and expansion is delayed until sufficient T cell help is generated. I will therefore argue here that, in response to an infection, initial clonal B-cell expansion and secretion of IgM occurs in a T cell-independent fashion (phase I) driven by the presence of antigen, and that secreted IgM serves as an autocrine growth factor at this time. B-cell-T-cell interaction occurs only during phase II of the response, thereby initiating the germinal center reaction, isotype switching and memory B-cell development. Hence, this model provides an explanation of how B-cell responses are induced rapidly in vivo at a time when T-cell help is rare. PMID- 11043777 TI - Follicular stromal cells and lymphocyte homing to follicles. AB - Follicular dendritic cells (FDCs), the best defined stromal cell subset within lymphoid follicles, play a critical role in presenting intact antigen to B lymphocytes. The discovery that many follicular stromal cells make B-lymphocyte chemoattractant (BLC), a CXC chemokine that attracts CXCR5+ cells, provides a basis for understanding how motile B cells come into contact with stationary FDCs. Here we review our work on BLC and discuss properties of BLC-expressing follicular stromal cells. We also review the properties of primary follicle and germinal center FDCs and suggest a model of FDC development that incorporates information about BLC expression. Finally, we consider how antigen recognition causes T and B lymphocytes to undergo changes in chemokine responsiveness that may help direct their movements into, or out of, lymphoid follicles. PMID- 11043778 TI - Regulation of humoral immune responses by CD21/CD35. AB - Before antigen-specific immunity arises, the complement system responds by activation through the classical and/or alternative pathways leading to the covalent deposition of complement fragments. Three models, not mutually exclusive, have been proposed to explain how these complement fragments interact with their receptors, CD21/CD35, to enhance humoral immune responses: i) CD21/CD35 retain and focus antigens for optimal presentation; ii) CD21/CD35 on B cells serve as enhancing co-receptors for B-cell antigen receptor (BCR) signaling; iii) CD21/CD35 regulate B-cell responses, by CD19 aggregation. The coreceptor model led us to predict that CD21/CD3 5 may lower the threshold of BCR affinity to diversify the repertoire of humoral immune responses, but surprisingly, CD21/CD3 5-deficient mice expressing a transgenic BCR with very low affinity (Kalpha approximately =1.2 x 10(5) M(-1)) for the (4-hydroxy-3 nitrophenyl)acetyl hapten generated significant antibody and germinal center responses to even low doses of antigens in alum. The magnitudes of these responses were much below those of normal controls but higher doses of antigens substantially rectified these deficits. Thus, while CD21/CD35 play important roles in humoral immunity, our observations provide little support to the hypothesis that CD21/CD35 promote clonal diversity in immune responses by helping recruit low-affinity B cells. PMID- 11043779 TI - Generation, expansion, migration and activation of mouse B1 cells. AB - We have studied the expansion, activation, homing and antibody production of B1 cells in two different mouse models. One is the HL transgenic mouse, which carries Ig genes encoding the anti-red blood cell autoantibody (4C8) and develops autoimmune hemolytic anemia by the activation of autoreactive B1 cells that escape from clonal deletion and expand in the peritoneal cavity (PEC). The other model is represented by alymphoplasia (aly) mice, which carry a point mutation in the gene encoding NF-kappaB-inducing kinase (NIK) and have drastically reduced immunoglobulin serum levels, in spite of their peritoneal cavity containing a large number of B1 cells. We have found that a) expression levels of the B-cell antigen receptor (BCR) influence the size of the B1 -cell compartment and efficiency of allelic exclusion and B2-cell deletion; b) antibody production of B1 cells is closely related with their migration from PEC to other lymphoid organs and is dependent on NIK; and c) infection, lipopolysaccharide stimulation, cytokine administration or T-cell activation by noncanonical antigens induces migration and differentiation of peritoneal B1 cells into antibody-producing cells. We describe a scenario where most of B1 and B2 differences are due to a distinct activation threshold of BCR and antigen repertoire. PMID- 11043780 TI - B-lymphocyte quiescence, tolerance and activation as viewed by global gene expression profiling on microarrays. AB - Self-tolerance is achieved by deleting or regulating self-reactive lymphocytes at a series of cellular checkpoints placed at many points along the developmental pathways to plasma cells and effector T cells. At each checkpoint, what are the molecular pathways that determine whether a lymphocyte remains quiescent, begins dividing, differentiates or dies? In splenic B cells, the decision between quiescence, tolerance by anergy, and activation provides a tractable setting to explore these issues by global gene expression profiling on DNA microarrays. Here we discuss the application of microarrays to illuminate a set of cell fate decisions that appear to be determined by summation of numerous small changes in expression of stimulatory and inhibitory genes. Many genes with known or predicted inhibitory functions are highly expressed in naive, quiescent B cells, notably the signal inhibitor SLAP and DNA-binding proteins of the Kruppel family (LKLF, BKLF, GKLF), Tsc-22, GILZ, Id-3, and GADD45. Activation of naive B cells, triggered by acute binding of antigen to the B-cell receptor, involves a rapid decrease in expression of these inhibitory genes. Promitotic genes are induced in parallel, including c myc, LSIRF/IRF4, cyclin D2, Egr-1 and Egr-2, as are the anti-apoptotic gene A1 and genes for the T-cell-attracting chemokines MIP-1alpha and beta. B-cell tolerance through the process of anergy, induced by chronic binding of self antigen, maintains expression of the inhibitory genes found in quiescent B cells and induces an additional set of inhibitory genes. The latter include inhibitors of signaling - CD72, neurogranin, pcp4 - and additional inhibitors of gene expression such as SATB1, MEF2C, TGIF and Nab-2. The effects of tolerance, the immunosuppressive drug FK506 and other modulators of calcium or MAPK signaling allow individual gene responses to be linked to different signal transduction pathways. The global molecular profiles obtained illustrate how quiescence and anergy are actively maintained in circulating B cells, how these states are switched to clonal expansion and how they could be better emulated by pro-tolerogenic drugs. PMID- 11043781 TI - Pressure-induced retention of the lysozyme on reversed-phase liquid chromatography. AB - This study investigated the effect of pressure on the retention behavior of a model protein, lysozyme, on octadecylsilica (C18) stationary phase under various equilibrium conditions. It is demonstrated that the retention time of the lysozyme was increased by as much as two to three times as the absolute pressure on the viewing window was increased from 23 to 318 bar. This pressure-induced retention was likely to be reversible and the corresponding volume change (deltaV = Vsta - Vmob) was found to be on the order of minus tens to hundreds of mL/mol. Moreover, the pressure-induced retention was also observed for a homologous series of hydrophobic poly-L-phenylalanine, which do not have the secondary structure, and the volume change was determined to be around minus 10 mL/mol per phenylalanine. Perturbations in solute ionization and conformational change are predicted to have a minor impact under the investigated conditions. It is believed that the pressure-induced shift of the equilibria regarding hydrophobic ad-desorption is the major cause of the observed increase of protein retention. About ten phenylalanine-equivalent residues on the lysozyme surface were involved in the hydrophobic association with the chromatographic ligands. PMID- 11043782 TI - Influence of the column radial heterogeneity on the determination of single component isotherms by the elution by characteristic point method. AB - The influence of the column radial heterogeneity on the determination of equilibrium isotherm data by the elution by characteristic point (ECP) method is studied using nondimensional numerical calculations and taking into account typical radial distributions of the mobile phase flow velocity and the column efficiency across a column. Overloaded elution peaks were calculated with the equilibrium-dispersive model as a function of four dimensionless parameters, the number of theoretical plates at the center of the column, the Langmuir equilibrium constant, the retention and the loading factors. The influence of the mass transfer resistances and the radial heterogeneity of the column on the ECP data was analyzed by comparing the true isotherm and the one estimated from the diffuse profile of overloaded peaks. The results provide information on the accuracy of the ECP method. The error made increases with increasing degree of radial heterogeneity. This error can be corrected by using the results of the nondimensional numerical calculations, allowing a further extension of the applicability of the ECP method. PMID- 11043783 TI - Approaches towards the quantitative analysis of peptides and proteins by reversed phase high-performance liquid chromatography in the absence of a pure reference sample. AB - A reversed-phase HPLC protocol for the quantitative analysis of peptides and proteins is presented. It is applicable to purified samples and potentially to crude biological extracts. The key feature is that an analytically pure reference sample of the analyte is not required because the extinction coefficient for the UV absorbance at 280 nm can be accurately estimated from the amino acid sequence. The concentration of a protein can therefore be calculated from the peak area relative to an internal standard. Sources of error and limitations of the method are systematically considered. Tryptophan containing peptides gave closer agreement to expected values than those with only tyrosine. It was found that analogous, previously used methods could not be directly applied to lower wavelengths. PMID- 11043784 TI - Determination of vitamin B12 in multivitamin tablets and fermentation medium by high-performance liquid chromatography with fluorescence detection. AB - A novel method for the determination of vitamin B12 by high-performance liquid chromatography with fluorescence detection is reported. The method was simple and highly sensitive with good precision. Vitamin B12 was analyzed by HPLC on a muBondapak C18 column (300x3.9 mm, 10 microm) with methanol-water (30:70) as mobile phase and fluorescence detection at 305 nm (with excitation at 275 nm). The calibration graph was linear from 1.000 to 100.0 ng ml(-1) for vitamin B12 with a correlation coefficient of 0.998 (n=6). The detection limit was 0.1 ng ml( 1). The method was successfully applied to the determination of vitamin B12 in vitamin B12 tablets, multivitamin tablets and fermentation medium. The recovery was from 94 to 102% and the relative standard deviation was in the range of 1.8 to 4.1%. PMID- 11043785 TI - Determination of chlorophacinone and diphacinone in commercial rodenticides by liquid chromatography-UV detection and liquid chromatography-electrospray ionization mass spectrometry. AB - This work describes a simple method for the determination and mass-spectrometric confirmation of the indanediones in commercial rodenticides. The sample is sonicated in methanol containing 2% formic acid and analyzed by liquid chromatography-UV detection. Once retention time and UV-Vis spectrum provide tentative identification, mass-spectrometric confirmation is obtained by analyzing a second aliquot by LC and electrospray ionization mass spectrometry (LC-ESI-MS). The extensive fragmentation of the indanedione molecule under MS/MS conditions provides sufficient structural information for positive identification on analyte levels as low as 20 ng on column. PMID- 11043786 TI - Configurational analysis of chiral acids as O-trifluoroacetylated (-)-menthyl esters by achiral dual-capillary column gas chromatography. AB - The simultaneous enantiomeric separation of 30 racemic acids including 24 hydroxy acids in a single analysis is described for the determination of their absolute configurations. It involves the conversion of each enantiomer into diastereomeric O-trifluoroacetylated (-)-menthyl ester for the direct separation by gas chromatography on achiral dual-capillary columns of different polarities, with subsequent identification and chiral discrimination by retention index (I) library matching. Among the acids studied, the enantiomers of 27 acids were discriminatively resolved on both non-polar DB-5 and the intermediate-polar DB-17 columns with resolution factors in the range of 0.7-7.7 and separation factors in the range of 1.002-1.021. Enantiomers of 3-hydroxybutyric and alpha methoxyphenylacetic acids were partially resolved on DB-5 (resolution factor of 0.9), but not resolved on DB-17, while the baseline resolution for 3 hydroxydecanoic acid and the minimal separation on the peak top (resolution factor less than 0.7) for 2-hydroxyglutaric acid were achieved on DB-17 but not on DB-5. The temperature-programmed I values measured on both columns were characteristic of each enantiomer and thus simple I matching with the reference values was useful in cross-checking for their chemical identification and the chiral discrimination as well. When applied to a clinical urine sample, the present method allowed positive identification of endogenous (S)-lactic acid and (S)-2-hydroxybutyric acid along with (R)-3-hydroxybutyric acid. PMID- 11043787 TI - Microwave-assisted derivatization of volatile carbonyl compounds with O (2,3,4,5,6-pentafluorobenzyl)hydroxylamine. AB - A method for the determination of carbonyl compounds, either directly from gaseous phase or following a volatilization from liquid or solid samples after trapping on Tenax TA is presented. Following solvent desorption, the carbonyls are derivatized using O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine. The reaction is accomplished in a microwave oven using closed vessels to minimize reaction time compared to conventional methodology. The solvent for the chemical reaction was selected according to the requirements of microwave energy interaction and solubility. After gas chromatographic separation of the corresponding oximes, they are detected using electron impact mass spectrometry in single ion monitoring mode. Quantification is carried out using internal standardization with 3-fluorobenzaldehyde, resulting in limits of detection in the ppm range following the calibration graph method. The optimized conditions provide for good recoveries and fast reaction rates for the volatile carbonyls studied so far. PMID- 11043788 TI - Miniaturised pressurised liquid extraction of polycyclic aromatic hydrocarbons from soil and sediment with subsequent large-volume injection-gas chromatography. AB - Analyte extraction is the main limitation when developing at-line, or on-line, procedures for the preparation of (semi)solid environmental samples. Pressurised liquid extraction (PLE) is an analyte- and matrix-independent technique which provides cleaner extracts than the time-consuming classical procedures. In the study, the practicality of miniaturised PLE performed in a stainless-steel cell, and combined with subsequent large-volume injection (LVI)-GC-MS was studied. As an example, the new system was applied to the determination of polycyclic aromatic hydrocarbons (PAHs) in soils and a sediment. Variables affecting the PLE efficiency, such as pressure and temperature of the extraction solvent and total solvent volume, were studied. Toluene was selected as extraction solvent and a total solvent volume of 100 microl was used for the 10 min static-dynamic PLE of 50-mg samples. Additional clean-up or filtration of the sample extracts was not required. Detection limits using LVI-GC-MS were below 9 ng/g soil for the 13 PAHs more volatile than indeno[1,2,3-cd]pyrene in real soil samples and the repeatability of the complete PLE plus LVI-GC-MS method for the analysis of the endogenous PAH was better than 15%. Comparison of PLE and Soxhlet or liquid partitioning extraction results for the analysis of non-spiked samples showed that the efficiency of PLE is the same or better than for the other two extraction methods assayed. PMID- 11043789 TI - Application of gas chromatography coupled to chemical ionisation mass spectrometry following headspace solid-phase microextraction for the determination of free volatile fatty acids in aqueous samples. AB - Gas chromatography coupled to positive and negative ion chemical ionisation mass spectrometry was evaluated for the determination of free volatile fatty acids (VFAs) from aqueous samples by headspace solid-phase microextraction. Negative ion chemical ionisation in the selected ion monitoring mode using ammonia as reagent gas provided acceptable sensitivity and the highest selectivity for the determination of C2-C7 fatty acids using a polydimethylsiloxane-Carboxen fibre. Detection limits in the range of 150 microg l(-1) for acetic acid and from 2 to 6 microg l(-1) for the remaining carboxylic acids were achieved. The reproducibility of the method was between 9 and 16%. The developed analytical procedure was applied to the analysis of VFAs in raw sewage. The absence of interfering peaks provided a more accurate determination of acetic, propionic, butyric and isovaleric acids than a similar analytical scheme but using a flame ionisation detector. PMID- 11043790 TI - Efficiency of pretreatment of aqueous samples using a macroporous strong anion exchange resin on the determination of nerve gas hydrolysis products by gas chromatography-mass spectrometry after tert.-butyldimethylsilylation. AB - A pretreatment procedure, using a macroporous strong anion-exchange resin (MSA) has been established for the determination of nerve gas hydrolysis products by gas chromatography-mass spectrometry (GC-MS) after tert.-butyldimethylsilyl (TBDMS) derivatization. Aqueous solutions of methylphosphonic acid (MPA) and three alkyl methylphosphonic acids (AMPAs) (ethyl, isopropyl and pinacolyl methylphosphonic acid), were retained on the MSA column, and then quantitatively eluted with 0.1 M hydrochloric acid. The neutralized column eluate was dried, and MPA and AMPAs were derivatized with N-methyl-N-(tert.-butyldimethylsilyl) trifluoroacetamide and analyzed by GC-MS. The column eluate was also analyzed in order to determine the exact hydrolysis product levels by capillary electrophoresis using borate and benzoate buffer (pH 6). The MSA pretreatment was examined for the clean-up of aqueous extracts of three types of soils and an aqueous solution containing 10% sucrose, which is regarded as model for a typical soft drink, after spiking with MPA and AMPAs. MPA and AMPAs were quantitatively recovered in the MSA eluate fraction from those samples, except for MPA from volcanic acid and alluvial soils. The yields of TBDMS derivatives were remarkably improved, compared with for which no pretreatment was used and also for those in which a strong cation-exchange resin was used. The achieved detection limits of MPA and AMPAs ranged from 0.12 to 0.18 microg/g of soil (S/N=3). The established MSA method was applied to the pretreatment of spiked sea water, two types of beverages, Pepsi Cola and canned coffee. Although the yields of TBDMS derivatives of MPA and AMPAs in sea water (in a range between 44 and 96%) and AMPAs in Pepsi Cola (in a range between 58 and 92%) were rather high, those for MPA in the Pepsi Cola (27%) and those for MPA and AMPAs in the canned coffee (in a range between 5 and 17%) were low. PMID- 11043791 TI - Monitoring organochlorine pesticides from landfill leachates by gas chromatography-electron-capture detection after solid-phase microextraction. AB - Landfill leachates contain significant amounts of organic carbon, nitrogen and heavy metals as well as other specific trace organic compounds like organochlorine pesticides. In this study a simple and reliable methodology was improved to detect organochlorine pesticides in leachate samples by using a previous solid-phase microextraction procedure [with a 100 microm poly(dimethylsiloxane) fiber] and chromatographic analysis by GC-electron-capture detection. The extraction time, temperature, ionic strength of the solution and sampling of the headspace were the parameters studied. Reproducibility achieved values below 20% RSD, and standard addition was used for pesticides confirmation. PMID- 11043792 TI - Polymer additive analysis by pyrolysis-gas chromatography. III. Lubricants. AB - Lubricants are widely used in thermoplastic polymers to increase the overall rate of processing or to improve surface release properties. Because of the low level of lubricants normally used in a polymer, it may not be possible to analyze the additive directly by common spectroscopic or thermal chemical techniques. However, lubricants as well as other additives in the polymer can be qualitatively analyzed by pyrolysis-gas chromatography (Py-GC) after extraction. In this work, several lubricants have been studied to demonstrate that Py-GC is a viable tool to investigate lubricants. The advantages of using Py-GC in the analysis of lubricant have also been discussed. PMID- 11043793 TI - Polymer additive analysis by pyrolysis-gas chromatography. IV. Antioxidants. AB - Antioxidants are important additives in polymers. Because of the low level of antioxidants normally used, they cannot be analyzed directly by common spectroscopic or thermal chemical techniques. However, antioxidants as well as other additives in polymers can be qualitatively analyzed by pyrolysis-gas chromatography (Py-GC) after separating the polymers and additives. In this study, several antioxidants have been investigated to demonstrate that Py-GC is a viable tool to analyze them. The advantages of using Py-GC in the analysis of antioxidants have also been discussed. PMID- 11043794 TI - Experimental study on moving neutralization reaction boundary created with the strong reactive electrolytes of HCl and NaOH in agarose gel. AB - In this paper, a moving neutralization reaction boundary (MNRB) is created with the strong reactive electrolytes of HCl and NaOH in agarose gel. The motions of the MNRB are investigated and compared with the predictions with the theory of the moving chemical reaction boundary (MCRB). The results show that, under appreciate experimental conditions, the experiments on the MNRB are exactly in coincidence with the predictions with the MCRB theory. Thus, the results excellently demonstrate that the MCRB theory is valid for the MNRB formed with the strong reactive electrolytes of HCl and NaOH. Additionally, it is, as discussed in this paper, imperative to develop a method to obtain ionic mobility at different temperatures and ionic strengths, in order to investigate the movements of the MCRB more efficiently. PMID- 11043795 TI - Separation of the molecular species of intact phosphatidylethanolamines and their N-monomethyl and N,N-dimethyl derivatives by high-performance liquid chromatography on a C8 column. AB - We have developed a gradient reversed-phase C8 high-performance liquid chromatography method for the separation of molecular species of phosphatidylethanolamines (PEs) and their N-monomethyl and N,N-dimethyl derivatives. This method uses a 40-min linear gradient of 88-100% methanol, containing ammonium hydroxide as silanol suppressing agent, and is suitable for metabolic studies using both UV detection at 205 nm and radioactivity flow detection. The elution order of a given PE is inversely related to the polarity of its fatty acid constituents. Lipid classes studied here containing the same fatty acyl chains elute in the order: PE-N,N-dimethyl or = 65 years. This paper reviews the available evidence on whether elderly influenza vaccination is worthwhile from a pharmacoeconomic point of view. A search on Medline and EMBASE resulted in a primary selection of approximately 100 studies on the pharmacoeconomics of influenza vaccination in the elderly. Further selection of studies to be included in the review was based on several criteria such as original research paper, cost-benefit or cost-effectiveness analysis. influenza vaccination in the elderly, and publication between 1980 and 1999. The 10 studies included in the final selection were evaluated regarding 3 main aspects: benefit-cost ratio and cost-effectiveness ratio; vaccine effectiveness; and relative costing of the vaccine. In general, differences in benefit-cost ratios could be explained by differences in effectiveness and relative costing of the vaccine. Considering the available pharmacoeconomic evidence, influenza vaccination of the elderly in western countries is an intervention with favourable cost-effectiveness in terms of net costs per life-year gained and even has cost-saving potential. In particular, influenza vaccination among elderly people at higher risk, such as the chronically ill elderly, is generally found to be cost saving. Relatively favourable cost-effectiveness among non-high-risk elderly justifies universal influenza vaccination of the elderly from a pharmacoeconomic point of view. PMID- 11043822 TI - The role of chemotherapy in prostate cancer. Minireview. AB - Hormonal therapy in disseminated prostate cancer is effective in 70-80% of patients and prolongs their lives of a mean 1-2 years. Sooner or later, androgen independence develops due to a multifactorial mechanism. A smaller part of patients may respond to second-line hormonal manipulations (antiandrogen withdrawal, adrenal enzymes synthesis inhibitors, corticosteroids). In hormone refractory disease only about 30% of patients would respond to chemotherapy. In the standard chemotherapy the mostly used cytotoxic agents are anthracyclines, platinum derivatives, vinca alkaloids and cyclophosphamide. However, combined chemotherapy is not more effective than monotherapy. Conventional chemotherapy may improve especially the quality of life. The median survival in chemotherapy patients (6-12 months) is not significantly longer when compared with the best supportive care. In recent years the main concern has been focused on new cytotoxic drugs and different combinations with hormonal agents. In Phase II studies the combinations of estramustine with oral etoposide, estramustine with taxanes and alternating weekly regimens (doxorubicin, ketoconazole/estramustine, vinblastine) show higher response rates (53-69% of patients with prostate specific antigen decline of more than 50%) and longer survival (13-19 months) than conventional chemotherapy. PMID- 11043823 TI - Retrovirus vector containing wild type p53 gene and its effect on human glioma cells. AB - A retroviral vector containing wild-type p53 tumor suppressor gene (wt-p53) under the control of viral LTR sequences was constructed and transfected into packaging cell line GP+envAm12. Virus producing single cell clone GP+envAm12/ p53clC8 (8 x 10(5) cfu/ml, determined on NIH 3T3 cells) was isolated and used to transfer wt p53 gene into human glioma cell lines in vitro. Decreased viability in p53 infected cells as compared to uninfected or empty virus infected cells was observed. PMID- 11043821 TI - Treatment and outcomes for elderly patients with small cell lung cancer. AB - It is estimated that approximately half of the 500 000 people diagnosed with lung cancer worldwide every year are aged >70 years. Thus, this disease represents a major problem in the elderly and one that will indeed increase as the median age of the population increases. For small cell lung cancer (SCLC), which accounts for approximately 20% of cases of lung cancer, the primary treatment is chemotherapy and in the majority of cases the primary aim is to control the disease which generally would have spread beyond the lungs at the time of presentation. A small number of 'standard' chemotherapy regimens (combined with radiotherapy for patients with limited disease) have been shown to improve survival and quality of life and are widely used. Much of the work investigating the relationship between age and treatment outcomes has been based on clinical trial data and may itself be inherently biased due to trial eligibility criteria excluding elderly patients. However, there is no good evidence that elderly patients fare worse with treatment than their younger counterparts in terms of response rates and survival. Nevertheless with increasing age comes increasing concomitant illnesses which may account for the widely observed increases in drug toxicity, and this may be the primary consideration in selecting the treatment option. Thus for many elderly patients, carboplatin/ etoposide may be the treatment of choice because it is perhaps the least toxic of the standard regimens. Whatever regimen is chosen, the key to treatment effectiveness seems to be to deliver the first 3 or 4 cycles without delay or dosage reduction. Although palliation of symptoms remains a major goal in the treatment of all patients with SCLC there is a dearth of data on whether elderly patients are equally well palliated as their younger counterparts. There is no good evidence that age per se should be a factor in deciding whether patients should receive standard treatment rather than a more gentle approach, and more elderly patients should be included in clinical trials. The key areas where more information is required regarding the treatment and outcomes of elderly patients with SCLC are the assessment of palliation, and comprehensive reviews of all patients diagnosed with the disease, not just those included in trials. PMID- 11043824 TI - Constitutive ubiquitination and degradation of c-myb by the 26S proteasome during proliferation and differentiation of myeloid cells. AB - Steady state levels of transcription factors play an important role in proliferation and differentiation of hematopoietic cells. The transcription factor c-Myb is frequently activated by retrovirus integration in murine and avian leukemias. Its deregulation has been also implicated in human acute and chronic leukemias and some other nonhematopoietic tumors. It is a short-lived protein, which is rapidly degraded by the 26S proteasome. Truncation at the carboxyl (COOH) terminus, which has occurred in some oncogenic forms ofc-Myb, results in the increased resistance to proteolysis. This stabilization correlates in vitro with less efficient ubiquitination. Here, we report the first evidence of post-translational modification of c-Myb by ubiquitin in vivo using HA-labeled ubiquitin. We also show that, in contrast to the unstable wild type or amino (NH2)-terminally truncated c-Myb form, stable carboxyl (COOH)-terminally truncated c-Myb is not targeted to degradation by covalent attachment of ubiquitin in vivo. In addition, following an analysis of subcellular fractionation of proteins from cells treated with a 26S proteasome inhibitor we were able to localize c-Myb exclusively in the nuclear compartment, suggesting the absence of a requirement for export to cytoplasm prior proteolytic processing. Furthermore, pulse-chase experiments of c-Myb protein isolated from interphase cells or cells synchronized in the G2/M or G1 phases of cell cycle did not reveal substantial cell cycle dependent differences in proteolytic processing by the 26S proteasome. Also, the demonstration that the half-life of c-Myb in myeloid progenitor M1 cells induced to differentiate along the monocytic pathway is the same as in undifferentiated cells suggested that proteolytic breakdown of c-Myb is a constitutive process during proliferation and differentiation. PMID- 11043825 TI - Approaches to identification of HNPCC suspected patients in Slovak population. AB - Patients with hereditary non-polyposis colorectal cancer (HNPCC) have a DNA mismatch repair defect (MMR) in their tumor tissue that results in instability of microsatellite DNA sequences (MSI). Thus, MSI analysis may effectively indicate this form of cancer that should be then proved by analysis of germline mutations in MMR genes. The aim of this study was to identify HNPCC suspected patients in the Slovak population by investigating microsatellite instability in colorectal tumor tissues. MSI was studied at 5-11 loci in matched tumor and normal DNA using radioactively labeled PCR products separated on sequencing gels. High microsatellite instability (MSI-H) was present only in patients younger than 50 years, in 100% of patients having two affected relatives by colorectal cancer and in 67% of patients with only one affected relative. In both groups of patients colorectal cancer was present in two successive generations. No MSI-H was found in the group of patients older than 50 years, even if they had positive family history for colorectal cancer. Among all markers used, the BAT26 mononucleotide repeat (100%), DI0S197 and D13S175 (62.5%) dinucleotide repeats were the most frequently altered in the tumor tissues. Retrospective analysis revealed that some of the patients having MSI-H tumors have had clinicopathological characteristics frequently reported to HNPCC. The family members of those patients with MSI-H are enrolled in preventive health care program until mutational analyses will enable to select carriers from non-carriers of mutated MMR genes. PMID- 11043826 TI - Mammary carcinogenesis induced in Wistar:han rats by the combination of ionizing radiation and dimethylbenz(a)anthracene: prevention with melatonin. AB - The primary cancer chemoprevention is an important topic of experimental oncology. We have analyzed the possible oncostatic properties ofmelatonin in a combined model of radiation plus chemocarcinogen-induced mammary carcinogenesis. Virgin female rats of Wistar:Han strain were continuously irradiated with daily dose 96 mGy of gamma rays up to 15 days. At the end of irradiation, between 52-60 postnatal days, 7,12-dimethylbenz(a)anthracene was administered by gavage, in three 10 mg/rat consecutive doses. A part of animals drank melatonin in a concentration 100 microg/ml of tap water, continuously from the beginning of irradiation and 26 weeks after its end. The aim of the experiment was to investigate the preventive effect of melatonin on mammary tumor patterns. Relatively low incidence of mammary tumors in the noninfluenced group was probably connected with generally very low sensitivity of Wistar:Han female rats to single dose of chemocarcinogen in mammary carcinogenesis induction. In our trial melatonin decreased markedly the volume of mammary tumors, but did not influence any other tumor characteristics. The chemopreventive effect of melatonin, derived from in vivo realized mammary carcinogenesis study in female Wistar:Han rats was limited. The cancer preventive properties of melatonin should be investigated in the future especially from the standpoint of susceptible strain, effective doses, and mode plus sufficient length of application. PMID- 11043827 TI - Repeated administration of carcinogen in critical developmental periods increases susceptibility of female Wistar: han rats to mammary carcinogenesis induction. AB - Analysis and knowledge of individual strain susceptibility of experimental animals to induction of carcinogenesis is important especially in regard to possibility of transfer of these facts to human pathology, first of all to chemopreventive projects. Our group (AHLERS et al. [1]) reported very low sensitivity of female Wistar:Han rats to induction of mammary carcinogenesis by 7,12-dimethylbenz(a)anthracene (DMBA) and by N-methyl-N-nitrosourea (NMU). The aim of this paper was to increase the sensitivity of females of this strain to mammary carcinogenesis induction by repeated administration of NMU in a dose 50 mg/kg of b.w. in critical periods: on 3-4 postnatal days, on 21 day (critical period for development of ductal parts of mammary gland) and between 50-55 days (maximal proliferation of whole gland). In comparison with 38% incidence of mammary tumors after the single dose and 65% incidence after 3 subsequent doses between 50-60 days, the combination of administration (only) on 21 day and between 50-55 postanatal days resulted in 88% incidence the sensitivity of animals reached the level of highly susceptible rat strains. The latency period was significantly increased in groups with NMU given on 3-4, 21 days and between 45-55 days respectively, on 21 day and between 45-55 days in comparison with control group (one dose of NMU). The tumor frequency per group and per animal in all groups with repeated NMU administration was significantly higher than that of control group. The volume of tumors was not influenced either by repeated carcinogen application or by time of its administration. These results expand the possibilities of analysis of carcinogen effects in individual periods of rat postnatal development. PMID- 11043828 TI - Chromosomal aberrations, sister chromatid exchanges and high frequency cells in young patients with neurofibromatosis 1 (NF1). AB - Chromosomal aberrations (CAs), sister chromatid exchanges (SCEs) and high frequency cells (HFCs) have been assessed in peripheral blood lymphocytes of 10 neurofibromatosis (NF1) patients and 10 healthy controls. In both groups, the spontaneous rates and the induced (bleomycin for CA and MMC for SCE) frequencies were analyzed. No differences between cells from NF1 patients and controls were observed with respect to spontaneous or bleomycin induced CA. Spontaneous or MMC induced SCE frequencies were also similar in NF1 patients and controls. HFCs, on the contrary, were statistically lower in NF1 patients. PMID- 11043829 TI - Sequential intermediate high-dose therapy with etoposide, ifosfamide and cisplatin for patients with germ cell tumors. AB - Intermediate high dose VIP (etoposide, ifosfamide, cisplatin) achieved comparable efficacy and improved tolerance in comparison with high-dose chemotherapy plus PBSC in poor risk germ cell tumors. The aim of this study was to confirm the effectivity and tolerance of this regimen in clinical practice. Twenty-five consecutive patients, 9 previously untreated with poor prognosis and 16 relapsed, were treated with 1.6 VIP or 1.9 VIP+PBSC. A relative dose intensity of 1.6 VIP was used in 14 patients and 11 patients received the intensity of 1.9 VIP. Clinical response was achieved in 56% of patients. Fifty-eight percent of patients have survived more than 1 year and 44% more than 2 years. No significant difference was noted between previously treated and untreated patients, as well as between the patients on 1.6 VIP and 1.9 VIP, with the exception of improved 1 year survival of patients on 1.9 VIP. One of four cisplatin-refractory patients achieved durable partial remission with a normal level of tumor markers. Serious non-hematological toxicity was rare. Myelotoxicity of 1.9 VIP was less serious in comparison with 1.6 VIP regimen, but the difference was not significant. Sequential intermediate high-dose therapy is an effective and tolerable regimen for patients with poor risk germ cell tumor as well as for relapsed patients. PMID- 11043830 TI - Carboplatin and cyclophosphamide in the treatment of metastatic seminoma. AB - Cisplatin-based chemotherapy is highly effective in advanced seminoma, but at the cost of a considerable toxicity. The response rate of carboplatin is comparable with cisplatin combinations but the relapse rate is higher. Our study assesses the efficacy and the toxicity of the combination of carboplatin and cyclophosphamide in patients with advanced seminoma. Nineteen consecutive patients received 6 cycles of intravenous cyclophosphamide 750 mg/m2 and carboplatin 350 mg/m2, repeated every 21 days. The overall objective response rate was 100%, 11 patients (58%) achieved a complete response and 8 patients (42%) showed a partial response. At median follow up of 4.2 years 3 patients (15%) relapsed. The 2-year disease-free survival and the overall survival are 72 and 94%, respectively. This outpatient treatment was well tolerated and the toxicity was mild. One patient had granulocytopenic fever and one patient had grade 3 cystitis. The combination therapy with carboplatin and cyclophosphamide is an effective and tolerable regimen in advanced seminoma. PMID- 11043831 TI - Double modulation of 5-fluorouracil by leucovorin and low-dose methotrexate in advanced colorectal cancer. AB - A phase II study was carried out to evaluate the efficacy and toxicity of a double biochemical modulation of 5-fluorouracil (5-FU) by methotrexate (MTX) and leucovorin (LV) in patients with advanced unresectable colorectal cancer. Forty two patients with measurable metastases of colorectal cancer were treated with 5 FU in daily doses of 600 mg/m2 given in a 6-hour intravenous (i.v.) infusion on days 1-5, LV 50 mg/m2 i.v. on days 2,3 and 5, and MTX 40 mg/m2 i.v. on days 1 and 4, every 4 weeks. Twenty-eight patients had a single metastatic site, eleven double, whereas three had more than two metastatic sites. Objective response (one complete response) was observed in 12 of 40 patients (30%) (95% confidence interval 16-48), stable disease in 19 patients (47%) and progression in 9 patients (23%). Overall median survival was 12 months. Median time to progression was 6 months. Treatment was generally well tolerated. The most frequent adverse reactions were stomatitis (38%), nausea and vomiting (35%), diarrhea (31%), leukopenia (18%), and plantar-palmar erythroderma (15%). The combination of 5-FU, LV and MTX seems to be an active regimen in advanced colorectal cancer. PMID- 11043832 TI - Long-term (15 years) results of extended field radiotherapy in Hodgkin's disease. AB - From 1975 to 1990, 214 patients with the pathological Stage IA, IB, IIA, IIB and IIIA of Hodgkin's disease were treated by supradiaphragmatic and/or infradiaphragmatic mantle technique. Complete remission was achieved in 70 patients (8%) by means of radiotherapy only. Partial remission was achieved in 9 patients (2%). The survival at 10 years was 86% and 15 years it was 66%. The most frequent late complications were hypothyreosis, Lhermitte's syndrome and radiation pneumonitis. PMID- 11043833 TI - Clinical stages of cutaneous malignant melanoma in Bulgaria. AB - The aim of the study was to investigate the distribution of the newly diagnosed cases with cutaneous malignant melanoma by clinical stages in Bulgaria over the period 1993-1995 as a reason for improving both mclanoma prevention and control. Over the period 827 new cases with cutaneous malignant melanoma are registered in the country. A representative sample of 671 cases has been taken. The cases with a localized melanoma (Stage I and II) were prevalent - 509 (75.8% of all studied cases). The thick melanomas (Stage IIB) were most frequently encountered among the primary lesions. They were 207 cases (30.8%). The thin melanomas (Stage IA) were only 41 (6.1%). The proportion of the cases with nodal and in-transit metastases (Stage III) -122 (18.2%) and the proportion of the cases with distant metastases (Stage IV) -40 (6.0%) were quite high. The analysis of the results shows that the cases with cutaneous malignant melanoma in Bulgaria are detected quite late. The cases with early-diagnosed melanoma are prevalent among women, young persons and urban population, and the cases with advanced melanoma are more frequent among men, persons older than 50 and rural population. PMID- 11043834 TI - Trends in the incidence of non-melanoma skin cancer in Slovakia, 1978-1995. AB - Non-melanoma skin cancer (NMSC) incidence in Slovakia in the period 1978-1995 was analyzed. A total of 38,629 microscopically confirmed NMSC cases (19,600 in males and 19,029 in females) were registered by the National Cancer Registry: 31,714 (82.1%) were basal cell carcinomas (BCC), 6,396 (16.6%) squamous cell carcinomas (SCC) and only 519 (1.3%) other NMSC. Age standardized rates of NMSC increased in the given period by 59.1% in males and 58.5% in females. The greatest increase was observed for BCC, 70.4% and 65.0% in males and females respectively, the smaller for SCC, 13.5 and 18.8%. In the period 1993-1995 age-standardized incidence rates per 100,000 were 38.0 for BCC, 6.7 for SCC and 45.5 for all NMSC in males and 29.2, 3.8 and 33.6, respectively in females. The observed marked increase of incidence with age was particularly pronounced for SCC. In both sexes, head and neck was the most common localization of BCC and SCC (84.2 to 74.7%), followed by trunk for BCC (17.0% in males and 10.8% in females) and by upper limbs for SCC (with 11.6% in males and 12.5% in females). Very fast increase of BCC incidence over time, its slower increase with age as compared to SCC incidence and body-site distribution suggest that BCC etiology is much more similar to melanoma etiology than SCC one. Registration of NMSC in relation to changes in possible risk factors (i.e. sun exposure/protection, ozone layer decrease) is important to study the mechanism of disease occurrence and to support public health interventions. PMID- 11043835 TI - Expression of Bcl-2 in dysplastic and neoplastic cervical lesions in relation to cell proliferation and HPV infection. AB - Expression of the bcl-2 gene has been shown to effectively confer resistance to programmed cell death in a variety of tumors. The bcl-2 proto-oncogene is involved in the development of human follicular lymphomas and also in a number of solid tumors such as carcinomas of prostate, breast, lung and GIT. The present study was designed to analyze the role of Bcl-2 expression in cervical intraepithelial squamous neoplasias (CIN) and cervical invasive carcinomas. Special attention was given to the association of Bcl-2 expression with the grade of the lesion, proliferative activity (expression of nuclear antigen of proliferative cells - PCNA) and human papillomavirus (HPV) DNA positivity. We examined tissue samples obtained from 86 women with varying degrees of cervical disease. Bcl-2 and PCNA were investigated using immunohistochemical staining and detection of HPV DNA was performed by hybridization in situ. Increased Bcl-2 expression was observed in advanced degrees of dysplasia and in carcinomas. We found a strong association between the presence of Bcl-2 in pathological epithelium with both the degree of dysplasia and the proliferative activity. We also observed a significant correlation between the amount of Bcl-2 positive lymphocytes infiltrating the lesions and the degree of disease. We, therefore, suggest that Bcl-2 expression in these lymphocytes may influence the antiviral or antitumor immune response. On the other hand we did not detect any significant correlation between the Bcl-2 oncoprotein and the presence of HPV. These results indicate that Bcl-2 may play an important role in the development of cervical cancer. PMID- 11043836 TI - Loss of heterozygosity on chromosome 5 in vicinity of the telomere in gamma radiation-induced thymic lymphomas in mice. AB - Analysis of the loss of heterozygosity at the D5Mit143 locus was done for thymic lymphlomas induced by gamma-irradiation of mice from two reciprocal backcrosses (BALB/c x CcS-13)F1 x BALB/c and (BALB/c x CcS-13)F1 x CcS-13. BALB/c mice are susceptible to gamma-ray induction of lymphomas. The CcS-13 strain is one of 20 CcS/Dcm (CcS) series of recombinant congenic strains, and the CcS-13 mice are resistant to gamma-radiation-induced lymphomas [1, 8]. Our preliminary tests show 50% (6/12) frequency of allelic loss at the D5Mit143 locus in thymic lymphomas induced by gamma-irradiation of the mice from (BALB/c x CcS-13)F1 x BALB/c backcross. Yet, in gamma-radiation-induced lymphomas from the backcross made in opposite direction, namely, (BALB/c x CcS-13)F1 x CcS-13, the analysis with the DSMit143 marker revealed low incidence of the loss of heterozygosity, 6.7% (15). The D5Mit143 locus resides in the distal part of chromosome 5, close to the telomere. Allelic loss of heterozygosity at the D5Mit143 locus showed strain specificity. In each case, the lost allele derived from the CcS-13 resistant strain. Our current results and previously done) linkage analysis [8] let us to suspect existence of a putative tumor suppressor gene for gamma-radiation-induced lymphoma at the region of murine chromosome 5. PMID- 11043837 TI - The expression of apoptosis-related proteins and the apoptotic rate in glial tumors of the brain. AB - Modern molecular biology methods allow a more precise analysis of the biological characteristics of tumors and, consequently, a more precise treatment plan. The determination of apoptotic rate and expression of apoptosis-related proteins belong among the important prognostic/diagnostic markers in many tumors. The validity of these factors had not yet been sufficiently analyzed in astroglial tumors. The aim of this work was therefore to study mutual relationships between apoptotic rate, expression of apoptosis-regulating proteins and some clinical and histopathological data. The TUNEL method was used for the determination of apoptosis in 44 astroglial tumor specimens. The percentage of TUNEL positive cells was expressed by the TUNEL index (TI). The TI data was compared with the immunohistochemically detected expression of proteins involved in apoptosis (BCL 2, FAS, FAS-L, and caspase 1), with grading, age, proliferative activity (assessed by PCNA expression analysis) and overall survival of patients. The statistical evaluation of results was done by two-way sample analysis of variance. We have demonstrated significantly higher values of both TI and expression of FAS-L and caspase 1 in low grade tumors, which were characterized by a longer survival, lower average age and a lower expression of PCNA. FAS-L expression correlated significantly with the expression of the caspase 1. No significant difference was found between the expression of BCL-2 and FAS. These results suggest that the determination of TI in astroglial tumors may be an important prognostic marker. The expression of FAS-L and caspase 1 in low grade astroglial tumors could indicate the increased readiness to apoptosis via the FAS/FAS-L cascade. PMID- 11043838 TI - Detection of Epstein-Barr virus in Hodgkin's lymphoma (patients in the Czech Republic). AB - The frequency of EBV demonstrated in patients with Hodgkin's lymphoma (HL) shows geographical variability. In the present study, we investigated the frequency of EBV in HL patients in the Czech Republic. The presence of EBV was determined by immunohistochemistry (IHC) with anti LMP-1 antibody and by in situ hybridization (ISH) method for EBERs. We studied 142 cases with HL. The age of patients ranged from 4 to 82 years. The male to female ratio was 1.2 (males 55.6%). In the series of 142 patients 47 (33%) positive cases were found. The incidence of EBV-positive results was significantly, higher in males than in females (70.2 vs. 29.7%, p = 0.023). Five patients were found in the age group below 10 years. They were positive with LMP-1 antibody and for EBERs in ISH method. The same results were discovered in two patients above the age of eighty. The most frequent histologic types of HL were nodular sclerosis (64 cases) and mixed cellularity (62 cases), respectively. The former type contained 16 EBV-positive cases (25%) and the latter 24 (38%) positive cases. The lymphocyte depletion type 2 (67%); lymphocyte rich type 5 (38%). EBV-positivity examined by ISH and IHC methods determined not only diagnostic Hodgkin cells and Reed-Sternberg cells but also small lymphocytes. In IHC method were small lymphocytes positive in 11 cases, more sensitive ISH revealed 32 positive cases. PMID- 11043839 TI - Application of cytogenetic endpoints and Comet assay on human lymphocytes treated with vincristine in vitro. AB - The genotoxic potential of vincristine is assessed on human peripheral blood lymphocytes following administration of the drug at a dose 0.0875 microg/ml by use of single cell gel electrophoresis - Comet assay (SCGE), analysis of structural chromosome aberrations (CA), micronucleus assay (MN) and sister chromatid exchange (SCE) analysis. In vitro treatment of human lymphocytes with vincristine was performed on cells in G0 phase, as well on lymphocyte cultures 24 hours after stimulation with mitogen phytohemagglutinine. For the Comet assay at 24, 48 and 72 h the treated cells were embedded in agarose on slides, lysed with alkaline lysis solution and exposed to an electric field. DNA migrated within the agarose and formed comets whose length depends on the amount of DNA damage. For the analysis of structural CA cells were grown on F-10 medium for 48 hours, and for MN and SCE analysis for 72 hours. The results on SCGE showed an increase in tail length compared to control both in cells treated in G0 and in cells treated 24 h after mitogen stimulation. The amount of DNA damage was higher in cells treated with vincristine 24 h after mitogen stimulation. Administered concentration of drug caused total inhibition of lymphocytes growth in 72-h cultures for MN and SCE analysis indicating strong microtubule distruptive effects of vincristine. Analysis of structural CA reveals chromatid breaks and acentric fragments as the main aberration types both in cells treated in G0 and in cells treated 24 h after mitogen stimulation. Number of these aberrations was higher in cells treated in G0 phase. Results obtained in this study by use of different cytogenetic endpoints confirmed that vincristine exhibits both aneugenic and clastogenic effects on human lymphocytes. PMID- 11043840 TI - Dexamethasone does not enhance antileukemic activity of cladribine in mice with leukemias L1210 and P388. AB - Combination of corticosteroids with new purine analogs such as cladribine 2 chlorodeoxyadenosine, (2-CdA) and fludarabine (FAMP) is controversial. The possibility of potentiation of antineoplastic activity of 2-CdA or FAMP by corticosteroids has not been documented so far. On the other hand, such combination may increase immunosuppression and the risk of infections. The aim of our study was to evaluate the influence of 2-CdA and dexamethasone (DEX) on the survival time of mice bearing lymphoid leukemias L1210 and P388. CD2F1 strain mice (132 male) were used in the experiment. The animals were injected i.p. on day 0 with 10(6) leukemic cells. The drugs were given on days 1-5 i.p. in the following concentrations: 2-CdA - 20 mg/kg, DEX 1.25 mg/kg, DEX 2.5 mg/kg, DEX 5 mg/kg, DEX 10 mg/kg, alone and in combination. The animals were observed daily for survival for a minimum of 30 days. The efficacy of the therapy against leukemia (defined as increase in lifespan - ILS) was assessed as the percentage of the median survival time (MST) of the treated group(t) to that of the control group(c): ILS(%) = (MSTt/MSTc) 100. The survival time of mice bearing L1210 or P388 leukemia treated with both drugs simultaneously was not longer than that of mice treated with either of drugs alone. Combination of 2-CdA and DEX in doses 5 and 10 mg/kg resulted in decrease of survival time of animals bearing P388 leukemia as compared with the control group without any treatment. Our study revealed that combination of 2-CdA with DEX in both leukemias is not more effective than 2-CdA alone. These results may indicate that routine addition of corticosteroids to purine analogs in the treatment of lymphoid malignancies is not warranted. PMID- 11043841 TI - Comparison of three in vitro assays at evaluation of IC50 of acetylsalicylic acid, ferrous sulfate, amitriptyline, methanol, isopropanol and ethylene glycol in human cancer cells HeLa. AB - Evaluation of the 50% inhibitory concentration (IC50) of acetylsalicylic acid, ferrous sulfate, amitriptyline, methanol, isopropanol and ethylene glycol was done on human cancer cells cultured in in vitro conditions. Three different in vitro assays were used in this study: the plating efficiency test, the microprotein test and the neutral red uptake test. Obtained results were evaluated by statistical methods. All used methods seem to be useful for screening a cytotoxic potential of the tested chemicals. The knowledge of cytotoxic effects of frequently used chemicals on mammalian cells is important not only for necessary in vitro genotoxicity and carcinogenicity studies but also for assessing the toxicity of chemicals to find out possible hazards to the human health. Results presented in this paper underline the usefulness of the wider methodological approach for the comparison of the different endpoints as well as a necessity for selection of a battery of in vitro cytotoxicity tests allowing to estimate the possible harmful effects of xenobiotics. PMID- 11043842 TI - Adjuvant clodronate therapy in patients with locally advanced breast cancer--long term results of a double blind randomized trial. Slovak Clodronate Collaborative Group. AB - Between March 1990 and May 1993 seventy three patients with previously untreated breast cancer, Stage III or IV without osseal metastases were randomized to sodium clodronate 1600 mg daily p.o.(arm A = 37 patients) or placebo (arm B = 36 patients) over 2 years, additionally to standard therapy. Ten patients were not evaluable for response because of short duration of therapy (less than 2 months). Bone metastases developed in 30% of patients in arm A and 23% patients in arm B. Median time to appearance of bone metastases was 13 months in arm A and 28 months in arm B. Non-bone metastases appeared in 48% patients in arm A and in 48% patients in arm B. Time to development of non-bone metastases was 20 months in arm A and 16 months in arm B. Five-year survival was 41% in arm A and 39% in arm B. There were no significant differences between the treated and control arms. PMID- 11043843 TI - The impact of radiotherapy on the incidence and time of occurrence of local recurrence in early-stage breast cancer after breast conserving therapy. AB - There is still little information on the delay of local recurrence after conservatively treated and irradiation breast cancer. To evaluate the impact of radiation therapy (RT) on the incidence and on the time of occurrence of ipsilateral breast tumor recurrence (IBTR), we reviewed the treatment results in 415 women with UICC Stage I or II unilateral breast cancer. All underwent breast conserving surgery (BCS) and full axillary dissection between 1983 and 1987. Out of them 309 patients were irradiated and 106 were not. The median dose of RT was 50 Gy in five weeks to the whole breast. Systemic therapy, when it was given, consisted of 6-cycles of CMF for node positive premenopausal women and 20 mg tamoxifen for three years for postmenopausal women. The median follow up time was 120 months in survivors. The 10-year actuarial IBTR rate was 36.6% for the nonirradiated and 9.1% for the irradiated women (p = 0.0000); 48.6% for patients treated with CMF and 4.2% for those treated with CMF plus RT (p = 0.0051); 29.0% for patients treated with tamoxifen and 7.9% for those treated with tamoxifen plus RT (p = 0.0318). The patient's age and the presence of an extensive intraductal component (EIC) were both highly associated with the likelihood of tumor recurrence in the treated breast. Patients under 41 years of age had an actuarial 10-year IBTR rate of 75% without RT and 17. 1% with RT (p = 0.0006). Women with an EIC positive tumor had an IBTR rate of 88.9% when RT was not given and 27.2% when RT was given (p = 0.0003). In invasive lobular cancer, irradiated patients had a IBTR rate of 2.3%, compared to 53.2% for nonirradiated patients (p = 0.0008). RT resulted in a significant delay in the appearance of IBTR (p = 0.0250) and the median time was increased by 20.0 months. We conclude that RT has the property of not only preventing but also delaying IBTR. In invasive lobular tumors the risk of IBTR is very high when RT is omitted, but BCS plus radiation therapy is effective treatment. Patients wih EIC positive tumor are at high risk of IBTR even when a median dose of 50 Gy is given to the whole breast. PMID- 11043844 TI - Influence of previous treatment on palliative effect of HDR brachytherapy in advanced esophageal cancer. AB - General effectiveness and influence of previous treatment on value of palliative HDR brachytherapy were assessed in 35 patients with advanced esophageal cancer treated from 1992 till 1997 with brachytherapy HDR (BT). Twelve of them were treated only with BT, II received previously chemoradiotherapy (CHTT), 12 teleradiotherapy (TT). BT appeared to be effective method of palliation. No significant differences in effectiveness of BT in analyzed groups were observed. Sever complications were observed in 9 cases (26%), and that in patients treated previously. Brachytherapy seems to be efficient after previous treatment, however, in this case, the risk of complications increases. PMID- 11043845 TI - Male breast cancer. Does the prognosis differ compared to female? AB - Due to the low incidence of breast cancer in males there are not many reports in the literature. In this study we analyzed results of treatment in 65 breast cancer males, who had been treated in one institution. Radical surgery was performed in 45 patients. Lymph node metastases were found in 25 patients (55.5%), the tumor was usually moderately differentiated (21 pts - 46.7%). Median survival after radical surgery was 73 months compared to 38 months for nonsurgical patients (p < 0.0001). In the group of males after radical surgery the results of 5-, 10- and 15-year survival rates were 69.8, 59.7 and 31.3% respectively. Comparable analysis of two subgroups of patients with favorable (T1 or T2, N0, grade I or II) and unfavorable (T3 or N+ or grade III) prognostic factors was also performed. In the first subgroup the 5-, 10- and 15-year survival rates were 90, 77.4 and 62%, compared to 61.8, 23.1 and 23. 1% for the second subgroup. The multivariate analysis showed grading and node status as the strongest parameters influencing survival. Relative risk of death was over 3 times higher for nodal metastases and near 3 times higher for high grade carcinomas (p < 0.01), compared to patients without metastases and low grade of tumor. Similar analysis was performed when 45 males were compared to 500 selected women, with similar clinical parameters (age, node status, grading). Again, data indicated grading and lymph node status as the strongest prognostic factors. It was not unlikely, that gender had some influence on prognosis, when relative risk of death for males was over 1.5 times higher than for females, but this result was not clearly significant (p < 0.1 ). The question, whether male breast cancer prognosis is worse then in female remains open. Multiinstitutional prospective studies are needed in this area. PMID- 11043846 TI - Enhancement of the photodynamic antitumor effect by streptococcal preparation OK 432 in the mouse carcinoma. AB - Biological response modifier antitumor effects are enhanced by the activation of the host defense mechanisms. We have investigated the antitumor effect of photodynamic therapy (PDT) and/or local administration of a biological response modifier, the streptococcal preparation OK-432, on transplanted NR-S1 mouse squamous cell carcinoma. Hematoporphyrin oligomers (20 mg/kg body weight) were used to photosensitize PDT. A pulsed Nd:YAG dye laser, tuned at 630 nm, was used as the light source. The laser power was 15 mJ cm(-2) pulse(-1), and the irradiation time was 40 min. The photosensitizer was injected intraperitoneally 48 h before laser irradiation. Where used, OK-432 was injected into the tumor either 3 h prior to PDT or immediately afterwards. The antitumor effects were evaluated 48 h after each protocol by (a) estimating the area of tumor necrosis (%) in hematoxylin/eosin-stained specimens, and (b) bromodeoxyuridine immunohistochemistry. Furthermore, the tumor sizes were evaluated 3, 7 and 10 days after each protocol, and the survival time after each protocol was evaluated as well. The antitumor effect of PDT was enhanced by administration of OK-432 3 h before PDT, whereas the administration of OK-432 immediately after PDT did not potentiate a PDT antitumor effect. Treatment with OK-432 alone had little effect on tumors. Photodynamic therapy in combination with local administration of OK 432 3 h before PDT is considered to be a useful treatment modality. PMID- 11043847 TI - CD40 activation enhances the magnitude of cellular immune responses against p53 but not the avidity of the effectors. AB - Engagement of CD40 on the surface of antigen-presenting cells (APC) has been shown to substitute for T cell help in activating APC to stimulate cytotoxic T lymphocytes (CTL). We explored whether this powerful non-specific signal could enhance the CTL response to a self epitope from a tumor-associated antigen. We immunized mice with a lipopeptide covering the H-2Kd-restricted epitope, amino acids 232-240 of murine wild-type p53, followed by treatment with an activating anti-CD40 monoclonal antibody. Anti-CD40 antibody, given subcutaneously or intravenously, significantly enhanced effector activity against targets pulsed with non-lipidated 232-240 nonamer epitope peptide, as assessed both by a CTL lysis assay and an enzyme-linked immunospot (ELISPOT) assay for interferon-gamma secreting cells. However, despite this enhancement, we could not detect activity against targets expressing p53 endogenously by either assay. This most likely reflects the low avidity of the effectors as determined by a titration of peptide on the target cells. The implications of this work for cancer immunotherapy based on specific responses directed against tumor-associated antigens are discussed. PMID- 11043848 TI - Induction of human cytotoxic T lymphocytes that preferentially recognise tumour cells bearing a conformational p53 mutant. AB - The tumour-suppressor gene p53 is pivotal in the regulation of apoptosis, and point mutations within p53 are the commonest genetic alterations in human cancers. Cytotoxic T lymphocytes (CTL) recognise peptide-MHC complexes on the surface of tumour cells and bring about lysis. Therefore, p53-derived peptides are potential candidates for immunisation strategies designed to induce antitumour CTL in patients. Conformational changes in the p53 protein, generated as a result of point mutations, frequently expose the 240 epitope, RHSVV (amino acids 212-217), which may be processed differently from the wild-type protein resulting in an altered MHC-associated peptide repertoire recognised by tumour specific CTL. In this study 42 peptides (37 overlapping nonameric peptides, from amino acids 193-237 and peptides 186-194, 187-197, 188-197, 263-272, 264-272, possessing binding motifs for HLA-A2) derived from the wild-type p53 protein sequence were assayed for their ability to stabilise HLA-A2 molecules in MHC class I stabilisation assays. Of the peptides tested, 24 stabilised HLA-A2 molecules with high affinity (fluorescence ratio >1.5) at 26 degrees C, and five (187-197, 193-200, 217-224, 263-272 and 264-272) also stabilised the complexes at 37 degrees C. Peptides 188-197, 196-203 and 217-225 have not previously been identified as binders of HLA-A2 molecules and, of these, peptide 217-225 stabilised HLA-A2 molecules with the highest fluorescence ratio. Peptide 217-225 was chosen to generate HLA-A2-restricted CTL in vitro; peptide 264-272 was used as a positive control. The two primary CTL thus generated (CTL-217 using peptide 217 225; and CTL-264 using peptide 264-272) were capable of specifically killing peptide-pulsed T2 or JY cells. In order to determine whether these peptides were endogenously processed and to test the hypothesis that mutants expressing different protein conformations would generate an alternative peptide repertoire at the cell surface, a panel of target cells was generated. HLA-A2+ SaOs-2 cells were transfected with p53 cDNA containing point mutations at either position 175 (R-->H) or 273 (R-->H) (SaOs-2/175 and SaOs-2/273). Two HLA-A2-negative cell lines, A431 and SKBr3, naturally expressing p53 mutations at positions 273 and 175 respectively, were transfected with a cDNA encoding HLA-A2. The results showed that primary CTL generated in response to both peptides were capable of killing SaOs-2/175 and SKBr3-A2 cells, which possess the same mutation, but not SaOs-2/273, A431-A2 or SKBr3 cells transfected with control vector. This suggests that these peptides are presented on the surface of SaOs-2/175 and SKBr3-A2 cells in a conformation-dependent manner and represent potentially useful target peptides for immunotherapy. PMID- 11043849 TI - Comparative delineation of T cell clonotypes in coexisting syngeneic B16 melanoma. AB - B16 is a murine melanoma of C57B1/6 origin, which rapidly develops as a tumor when inoculated into syngeneic immunocompetent hosts. Nevertheless, B16 tumors are considered to be immunogenic since tumor regression can be induced by means of immunotherapeutic intervention. Furthermore, B16 melanoma cells express several melanoma-associated antigens that may serve as targets for autologous T cells. To study the in vivo T cell response against B16, with particular emphasis on diversity and systemic involvement, we examined the spectra of T cell clonotypes in coexisting B16 melanoma lesions in C57B1/6 mice. Three tumors from each animal (n = 8) were examined for the presence of clonotypic T cells using the highly sensitive T cell receptor (TCR) clonotype mapping technology. Systematic analysis of the TCRB variable regions 1-16 revealed from 19 to more than 30 clonotypic TCR transcripts in each tumor. To study intra- and inter individual variations in the T cell response further, more than 600 clono-typic TCR transcripts were compared for sequence identity. Overall, approximately 2% of the T cell clonotypes were detected in more than one tumor from the same animal. Furthermore, none of the detected clonotypes was present in more than one animal, arguing against recurrent or "public" T cell responses against B16 melanoma. Our data strongly suggest that anti-melanoma T cell responses in this murine model encompass mainly localized T cells, and that systemic involvement is limited. PMID- 11043850 TI - Inhibition of interleukin-10 (IL-10) production from MOPC 315 tumor cells by IL 10 antisense oligodeoxynucleotides enhances cell-mediated immune responses. AB - Interleukin-10 (IL-10) has both inhibitory and stimulatory effects on diverse cell types of the immune system. It inhibits the antigen-presenting capacity of monocytes/macrophages and stimulates T cell proliferation. Although many tumors spontaneously release IL-10, the physiological relevance of this phenomenon to the in vivo antitumor immune response is not known. To elucidate the physiological role of tumor-released IL-10, we used IL-10-specific antisense oligodeoxynucleotides (AS-ODN) for the inhibition of IL-10 production from the tumor cells. Incubation of MOPC 315 plasmacytoma with IL-10 AS-ODN in vitro resulted in inhibition of IL-10 production and also in enhancement of the expression of major histocompatibility complex (MHC) class I, MHC class II, and B7-1 molecules. MOPC 315 cells incubated with IL-10 AS-ODN (MOPC-IL10AS) for 16 h in vitro showed reduced tumorigenicity in Balb/c mice. The mice implanted with MOPC-IL10AS effectively rejected the tumor graft, and showed strong cytotoxic T lymphocyte (CTL) activity against the parental MOPC 315 cells. In addition, MOPC IL10AS were more effective as stimulator cells in mixed lymphocyte/ tumor cell culture, and as target cells in a CTL assay. These results imply that IL-10 spontaneously released from MOPC 315 cells inhibits their immunogenicity and that the inhibition of IL-10 production by IL-10 AS-ODN may be a way to enhance the host cellular antitumor immune response. PMID- 11043851 TI - A bispecific single-chain antibody directed against EpCAM/CD3 in combination with the cytokines interferon alpha and interleukin-2 efficiently retargets T and CD3+CD56+ natural-killer-like T lymphocytes to EpCAM-expressing tumor cells. AB - Cytokine-induced killer cells (CIK), generated in vitro from peripheral blood mononuclear cells (PBMC) by addition of interferon gamma (IFNgamma), interleukin 2 (IL-2), IL-1 and a monoclonal antibody (mAb) against CD3, are highly efficient cytotoxic effector cells with the CD3+CD56+ phenotype. In this study, we evaluated whether the cytotoxicity of these natural-killer-like T lymphocytes against the colorectal tumor cell line HT29 can be enhanced by the addition of a bispecific single-chain antibody (bsAb) directed against EpCAM/CD3. For determination of bsAb-redirected cellular cytotoxicity we used a new flow cytometric assay, which directly counts viable tumor cells and can assess long term cytotoxicity. We found that this bsAb induced distinct cytotoxicity at a concentration above 100 ng/ml with both PBMC and CIK at an effector-to-target cell ratio as low as 1:1. CIK cells revealed higher bsAb-redirected cytotoxicity than PBMC. Cellular cytotoxicity appeared after 24 h whereas PBMC showed the highest bsAb-redirected cytotoxicity after 72 h. The addition of the cytokines IL 2 and IFNalpha but not granulocyte/macrophage-colony-stimulating factor enhanced bsAb-redirected cytotoxicity of both PBMC and CIK. When the bsAb was combined with the murine mAb BR55-2, which recognizes the Lewis(Y) antigen, bsAb redirected cytotoxicity was partly augmented, whereas murine mAb 17-1A, which binds to EpCAM as well, slightly suppressed bsAb-redirected cytotoxicity induced by the bsAb. We conclude that CIK generated in vitro or in vivo combined with this new EpCAM/CD3 bsAb and the cytokine IL-2 should be evaluated for the treatment of EpCAM-expressing tumors. PMID- 11043852 TI - Synergy between interleukin-2 and prothymosin alpha for the increased generation of cytotoxic T lymphocytes against autologous human carcinomas. AB - Peripheral blood mononuclear cells (PBMC) from cancer patients were cultured in vitro with irradiated autologous tumor cells isolated from malignant effusions (mixed lymphocyte tumor cultures, MLTC) and low-dose (50 IU/ml) recombinant interleukin-2 (IL-2). The combination of IL-2 and prothymosin alpha (ProTalpha) resulted in a greater PBMC-induced response to the autologous tumor than that brought about by IL-2 alone. In particular, ProTalpha specifically enhanced the CD4+ T-cell-mediated proliferation against the autologous tumor. CD4+ T cells seemed to recognize tumor antigens presented by HLA-DR molecules expressed on the autologous monocytes, since preincubation of the latter with an anti-HLA-DR monoclonal antibody (mAb) abrogated the response. In addition, MLTC set up with IL-2 and ProTalpha also generated more MHC-class-I-restricted cytotoxic T lymphocytes (CTL) against the autologous tumor than did MLTC set up with IL-2 alone. The MLTC-induced CTL contained high levels of cytoplasmic perforin and their development was strictly dependent on the presence of both autologous CD4+ T cells and monocytes. In the absence of either population there was a strong impairment of both proliferative and cytotoxic responses which was not restored by the presence of ProTalpha. In contrast, when both cell populations were present, ProTalpha exerted optimal enhancement of CD4+ T cell proliferation, which was associated with potentiated CTL responses. Our data emphasize the role of ProTalpha for the enhancement of IL-2-induced CTL responses against autologous tumor cells. Such responses require collaborative interactions between CD4+, CD8+ T cells and monocytes as antigen-presenting cells. Our data are relevant for adoptive immunotherapeutic settings utilizing IL-2 and ProTalpha-induced autologous-tumor-specific CTL. PMID- 11043853 TI - Control of early development of the pancreas in rodents and humans: implications of signals from the mesenchyme. AB - During the last few years, progress has been made in the control of pancreatic development. Many transcription factors have been described in the pancreas and a genetic approach has been used to define their role in pancreatic development. Pancreatic development depends on mesodermic signals, with the initial steps controlled by signals from the notochord that is in close contact with the dorsal endoderm of the gut fated to become pancreas. Later signals from the mesenchyme that surrounds the embryonic pancreatic epithelium regulate the proliferation of immature pancreatic epithelial cells and their differentiation into endocrine or exocrine tissue. This review discusses recent data on the role the signals from the mesenchyme have in the development of the pancreas in rodents and humans. PMID- 11043854 TI - Use of cod liver oil during pregnancy associated with lower risk of Type I diabetes in the offspring. AB - AIMS/HYPOTHESIS: To test whether cod liver oil or vitamin D supplements either taken by the mother during pregnancy or by the child in the first year of life is associated with lower risk of Type I (insulin-dependent) diabetes mellitus in children. METHODS: We carried out a population-based case control study in Vest Agder county of Norway, evaluating the use of supplements by a mailed questionnaire. We received responses from 85 diabetic subjects and 1,071 control subjects. Odds ratios (OR) with 95% confidence intervals (CI) were estimated using logistic regression analyses. RESULTS: When mothers took cod liver oil during pregnancy their offspring had a lower risk of diabetes. The unadjusted OR was 0.30, 95% CI: (0.12 to 0.75), p = 0.01. This association changed very little and was still significant after adjusting for age, sex, breastfeeding and maternal education. Mothers taking multivitamin supplements during pregnancy [adjusted OR= 1.11, 95% CI: (0.69 to 1.77)], infants taking cod liver oil in the first year of life [adjusted OR = 0.82, 95 % CI: (0.47 to 1.42) and the use of other vitamin D supplements in the first year of life [adjusted OR = 1.27, 95 % CI: (0.70 to 2.31)] was not [corrected] significantly associated with the risk of diabetes. CONCLUSION/INTERPRETATION: We found that cod liver oil taken during pregnancy was associated with reduced risk of Type I diabetes in the offspring. This suggests that vitamin D or the n-3 fatty acids eicosapentaenoic acid and docosahexaenoic acid in the cod liver oil, or both, have a protective effect against Type I diabetes. PMID- 11043855 TI - The influence of improved glycaemic control with insulin and sulphonylureas on acute phase and endothelial markers in Type II diabetic subjects. AB - AIMS/HYPOTHESIS: Improved glycaemic control might reduce both microvascular and macrovascular complications of Type II diabetes (non-insulin-dependent) mellitus. To explore such possible mechanisms, we investigated the effects of intensive treatment on markers of endothelial dysfunction and of acute phase activation, using both sulphonylureas and insulin. METHODS: In a randomised cross-over study we gave sulphonylureas or insulin each for a period of 16 weeks to 22 poorly controlled Type II diabetic subjects who were being treated by diet. There was a 4 week washout period between each treatment. Subjects were studied at baseline and at the end of each treatment. RESULTS: Treatment with sulphonylureas and insulin resulted in similar improvements in glycaemic control (glycated haemoglobin, baseline: 11.8 [(SD 2.2)%; after sulphonylureas: 8.6 (1.2)%,p < 0.001; after insulin: 8.6 (1.2)%, p < 0.001] and in insulin sensitivity ?metabolic clearance rate of glucose, baseline: median 1.75 [interquartile (IQ) range 1.41, 2.27] ml x kg(-1) x min(-1); after sulphonylureas: 2.41 (1.82, 3.01) ml x kg(-1) x min(-1), p = 0.001; after insulin: 2.23 (1.92, 2.75) ml x kg(-1) min(-1), p = 0.027?. There were no significant changes in concentrations of endothelial markers von Willebrand factor, cellular fibronectin, thrombomodulin, tissue plasminogen activator, soluble E-selectin or soluble intercellular adhesion molecule-1 or in urinary albumin excretion rate after either treatment period. Concentrations of C-reactive protein were not significantly influenced by sulphonylureas but fell after insulin [baseline: median 4.50 (IQ range 1.37, 6.44) microg x ml(-1); sulphonylureas: 2.69 (0.88, 9.65) microg x ml(-1) (p = 0.53); insulin: 2.07 (1.16, 5.24) microg x ml(-1) (p = 0.017)]. There were, however, no significant effects of either treatment on circulating concentrations of fibrinogen (p = 0.28-0.34) or of the proinflammatory cytokines interleukin-6 or tumour necrosis factor-alpha (p = 0.65-0.79). CONCLUSION/INTERPRETATION: Markers of endothelial dysfunction and concentrations of proinflammatory cytokines in Type II diabetes are not influenced by improved glycaemic control over 16 weeks. Improved metabolic control with insulin could, however, be associated with reduced concentrations of the acute phase marker C-reactive protein. PMID- 11043856 TI - Impaired phosphorylation and insulin-stimulated translocation to the plasma membrane of protein kinase B/Akt in adipocytes from Type II diabetic subjects. AB - AIMS/HYPOTHESIS: To examine protein kinase B/Akt distribution and phosphorylation in response to insulin in different subcellular fractions of human fat cells from healthy subjects and subjects with Type II (non-insulin-dependent) diabetes mellitus. METHODS: We prepared subcellular fractions of plasma membranes (PM), low density microsomes and cytosol and examined gene and protein expression as well as serine and threonine phosphorylation in response to insulin. RESULTS: Protein kinase B/Akt mRNA as well as total protein kinase B/Akt protein in whole cell lysate and cytosol were similar in both groups. Insulin increased protein kinase B/Akt translocation to the the plasma membrane about twofold [(p < 0.03) in non-diabetic cells but this effect was impaired in diabetic cells (approximately 30%; p > 0.1)]. In both groups, protein kinase B/Akt threonine phosphorylation considerably increased in low density microsomes and cytosol whereas serine phosphorylation was predominant in the plasma membrane. Phosphatidylinositol-dependent kinase 1, which partially activates and phosphorylates protein kinase B/Akt on the specific threonine site, was predominant in cytosol but it was also recovered in low density microsomes. Serine phosphorylation in response to insulin was considerably reduced (50-70 %; p < 0.05) in diabetic cells but threonine phosphorylation was less reduced (approximately 20%). Wortmannin inhibited these effects of insulin supporting a role for PI3-kinase activation. CONCLUSION/INTERPRETATION: Insulin stimulates a differential subcellular pattern of phosphorylation of protein kinase B/Akt. Furthermore, insulin-stimulated translocation of protein kinase B/Akt to the plasma membrane, where serine phosphorylation and full activation occurs, is impaired in Type II diabetes. Threonine phosphorylation was much less reduced. This discrepancy may be related to differential activation of phosphatidylinositol 3-kinase in the different subcellular compartments and phosphatidylinositol-dependent kinase 1 having high affinity for phosphatidylinositol phosphate 3. PMID- 11043857 TI - The impaired renal vasodilator response attributed to endothelium-derived hyperpolarizing factor in streptozotocin--induced diabetic rats is restored by 5 methyltetrahydrofolate. AB - AIMS/HYPOTHESIS: Endothelial dysfunction contributes to the development of diabetic vascular complications. A better understanding of the pathophysiology of endothelial dysfunction in diabetes could lead to new approaches to prevent microvascular disease. METHODS: Endothelium-dependent and endothelium-independent vasodilator responses were investigated in the renal microcirculation of streptozotocin-induced diabetic rats. We measured renal blood flow changes with an electromagnetic flow probe. In addition, the responses of the different segments of the renal microcirculation were evaluated with videomicroscopy using the hydronephrotic kidney technique. Because endothelial cells release different relaxing factors (nitric oxide, prostacyclin and an unidentified endothelium derived hyperpolarizing factor), responses to acetylcholine were measured before and after treatment with the nitric oxide synthase inhibitor L-NG-nitroarginine methylester HCI (L-NAME) and the cyclooxygenase inhibitor indomethacin. We evaluated with the effect of 5-methyltetrahydrofolate, the active form of folate, on the responses. RESULTS: The L-NAME- and indomethacin-resistant vasodilation to intra-renal acetylcholine was significantly reduced in the diabetic compared with control rats, suggesting impaired endothelium-derived hyperpolarizing factor mediated vasodilation. The responses to the nitric oxide donor (Z)-1-[-2 (aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-i um-1,2-diolate (DETA-NONOate) and to the K+-channel opener pinacidil were similar in diabetics and controls, indicating intact endothelium-independent vasodilator mechanisms. The contribution of endothelium-derived hyperpolarizing factor to vasodilation induced by acetylcholine was greatest in the smallest arterioles. In diabetic rats, the response to acetylcholine was increasingly impared as vessel size decreased. Defective vasodilation in diabetic kidneys was rapidly normalized by 5 methyltetrahydrofolate. CONCLUSION-INTERPRETATION: Endothelium-derived hyperpolarizing factor-mediated vasodilation is impaired in the renal microcirculation of diabetic rats, in particular in the smallest arteries. Treatment with folate restores the impaired endothelial function in diabetes. PMID- 11043858 TI - Contribution of visceral obesity to the deterioration of the metabolic risk profile in men with impaired glucose tolerance. AB - AIMS/HYPOTHESIS: Impaired glucose tolerance is associated with metabolic alterations which increase cardiovascular disease risk. The contribution of hyperglycaemia to this increased risk is, however, not clear. Abdominal obesity is often observed in subjects with impaired glucose tolerance; our objective was therefore to find the contribution of visceral adipose tissue to the deterioration of the metabolic risk profile noted in subjects with impaired glucose tolerance. METHODS: We studied 284 men with a normal glucose tolerance and 66 men with impaired glucose tolerance which was defined as a glycaemia between 7.8 and 11.1 mmol/l 2 h after a 75-g glucose load. RESULTS: Men with impaired glucose tolerance had more visceral adipose tissue and higher concentrations of plasma glucose and insulin in the fasting state and following a 75-g oral glucose load than men with a normal glucose tolerance. They also had higher concentrations of plasma cholesterol, triglycerides, apolipoprotein B and lower concentrations of HDL-cholesterol as well as higher cholesterol:HDL cholesterol ratios than men with a normal glucose tolerance. The two groups of men were then compared after a statistical adjustment for the amount of visceral adipose tissue. Although men with impaired glucose tolerance still had higher fasting plasma glucose and insulin concentrations after the adjustment for visceral adipose tissue, differences in all the variables of the lipid lipoprotein profile were eliminated. CONCLUSION/INTERPRETATION: Visceral adipose tissue accumulation is an important factor in the deterioration of the plasma lipid-lipoprotein noted in men with impaired glucose tolerance. PMID- 11043859 TI - Fatty acids modulate protein kinase C activation in porcine vascular smooth muscle cells independently of their effect on de novo diacylglycerol synthesis. AB - AIMS/HYPOTHESIS: Diabetes-induced activation of protein kinase C has been associated with the development of vascular complications. Elevated de novo diacylglycerol synthesis has been postulated to underlie this protein kinase C activation. Diabetes also increases the circulating concentrations of non esterified fatty acids, which are immediate precursors of diacylglycerol through the de novo pathway. We hypothesized that increased fatty acids contribute to de novo diacylglycerol synthesis and activation of protein kinase C in vascular cells. METHODS: Primary cultures of porcine carotid smooth muscle cells were exposed to fatty acids, bound to albumin in physiologic ratios. Diacylglycerol and triacylglycerol were measured in extracts of these cells. Protein kinase C activation was measured as membrane translocation with isoform-specific antibodies. RESULTS: Saturated fatty acids caused considerable accumulation of diacylglycerol through de novo synthesis. Unsaturated fatty acids increased triacylglycerol, but not diacylglycerol. Platelet-derived growth factor activated the alpha, epsilon and zeta protein kinase C isoforms. Activation of the alpha and zeta isoforms was amplified by oleate pretreatment but inhibited by palmitate. In the absence of growth factor stimulation, neither palmitate nor oleate had any effect on the membrane/cytosol distribution of any protein kinase C isoform. CONCLUSION/INTERPRETATION: Saturated fatty acids elicited de novo diacylglycerol synthesis in vascular smooth muscle cells without activating protein kinase C. Effects of fatty acids on protein kinase C activation by platelet-derived growth factor did not correlate with the effects on de novo diacylglycerol synthesis. These results indicate that de novo diacylglycerol synthesis is, by itself, insufficient to activate protein kinase C. PMID- 11043860 TI - The HIV-1 protease inhibitor indinavir impairs insulin signalling in HepG2 hepatoma cells. AB - AIMS/HYPOTHESIS: Patients treated with human immunodeficiency virus-1 protease inhibitors often develop impaired glucose tolerance or diabetes, most likely due to an induction of insulin resistance. We therefore investigated whether the protease inhibitor indinavir alters insulin signalling. METHODS: We incubated HepG2 cells for 48 h without or with indinavir (100 micromol/l). Subsequently 125I-insulin binding to the cells and the effects of insulin stimulation on insulin-receptor substrate-1-phosphorylation, association of phosphatidylinositol 3-kinase with insulin-receptor substrate-1 and Akt-Thr308-phosphorylation were measured. RESULTS: In cells not exposed to indinavir, insulin (100 nmol/l) led to rapid increases of insulin-receptor substrate-1-phosphorylation, association of phosphatidylinositol 3-kinase with insulin-receptor substrate-1 and Akt phosphorylation during the first 75 s, followed by subsequent decreases. In indinavir-treated cells, these insulin-stimulated increases during the first 75 s were reduced by 30-60% and this was not associated with alterations in cell number or viability, insulin binding to the cells or cellular insulin-receptor substrate-1-content. CONCLUSION/INTERPRETATION: Effects of indinavir on initial insulin signalling could cause, or contribute to, the metabolic effects of human immunodeficiency virus-1 protease inhibitors. PMID- 11043861 TI - Fas ligand-mediated mechanisms are involved in autoimmune destruction of islet beta cells in non-obese diabetic mice. AB - AIMS/HYPOTHESIS: A mechanism implicated in pancreatic islet beta-cell destruction in autoimmune diabetes is the binding of the Fas ligand (FasL) on T cells to Fas receptors on beta cells, causing their destruction. Evidence for this mechanism is, however, controversial. The aim of this study was to find whether the Fas ligand contributes to beta-cell death in autoimmune diabetes. METHODS: We transplanted syngeneic islets under the renal capsule in non-obese diabetic (NOD) mice and treated the mice with a neutralizing monoclonal antibody to the Fas ligand. Survival of beta cells in islet grafts and phenotypes of graft infiltrating cells were investigated. RESULTS: We found 58% (7 of 12) of mice treated with anti-Fas ligand antibody were normoglycaemic at 30 days after islet transplantation compared with none (0 of 9) of the mice treated with control antibody. Immunohistochemical analysis of islet grafts showed that infiltration of leucocytes (CD4+ T cells, CD8+ T cells, macrophages and neutrophils) and apoptosis of beta cells in the grafts was significantly decreased in mice treated with anti-Fas ligand antibody. Expression of proinflammatory cytokines (interleukin 1 alpha, tumour necrosis factor alpha and interferon gamma) was not different in islet grafts of mice treated with anti-Fas ligand and control antibodies. CONCLUSION/INTERPRETATION: These findings indicate that Fas ligand mediated mechanisms play a major part in promoting leucocytic infiltration of islets and beta-cell destruction in autoimmune diabetes. PMID- 11043862 TI - Trafficking of non-regulated secretory proteins in insulin secreting (INS-1) cells. AB - AIMS/HYPOTHESIS: Sorting of proinsulin to the regulated secretory pathway of pancreatic beta cells and retention of insulin in dense-core granules of this pathway is remarkably efficient. To monitor the specificity of these events, the secretion of two exogenous secretory proteins not known to carry information for sorting or retention in the regulated pathway was investigated in INS-1 cells. METHODS: SEGFP, a fusion protein consisting of a signal peptide N-terminal to EGFP (mutant green fluorescent protein with enhanced fluorescence) and secreted alkaline phosphatase (SEAP) were expressed in INS-1 cells by transfection and by infection with recombinant adenovirus, respectively. Secretion of SEGFP was monitored by quantitative western blotting and that of SEAP by its activity. RESULTS: Secreted alkaline phosphatase showed high basal secretion (6.6% total) but only modest (3.6-fold) stimulation of secretion by secretagogues, in keeping with secretion largely through the constitutive pathway. By contrast SEGFP had a secretory pattern similar to insulin, with low basal secretion (0.8% total) and 16-fold stimulation by secretagogues. Granular localization of SEGFP was confirmed by high resolution electron microscopy immunocytochemistry. Pulse-chase experiments indicated retention of SEGFP in granules at least 24 h after synthesis. The secretory SEGFP, but not cytosolic EGFP, formed disulphide-linked oligomers. This could be implicated in its regulated secretion. CONCLUSION/INTERPRETATION: These data indicate that in INS-1 cells SEGFP, but not SEAP, is unexpectedly handled as a regulated secretory protein and stored along with insulin in granules. This raises questions about the specificity and mechanism of the sorting of proteins to granules in INS-1 cells or their retention therein or both. PMID- 11043863 TI - Expression of peroxisome proliferator-activated receptor gamma (PPARgamma) in normal human pancreatic islet cells. AB - AIMS/HYPOTHESIS: Thiazolidinediones are reported to improve pancreatic islet morphology and beta-cell function in rodents, supporting the hypothesis of a direct action of thiazolidinediones on endocrine islet cells. In this study we examined the expression of the peroxisome proliferator-activated receptor gamma, a nuclear receptor that is activated by naturally occurring fatty acids and synthetic thiazolidinediones, in normal human endocrine pancreatic cells. METHODS: Human islets were isolated from pancreata harvested in ten brain-dead lean non-diabetic adult donors. We analysed the gene and protein expression of the human peroxisome proliferator-activated receptor gamma and evaluated the effects of peroxisome proliferator-activated receptor gamma agonist on insulin secretion in human islet preparations. RESULTS: The RT-PCR carried out on total RNA from four distinct human islet preparations demonstrated the presence of peroxisome proliferator-activated receptor gamma mRNA. Western blot analysis showed the consistent expression of peroxisome proliferator-activated receptor gamma protein. Peroxisome proliferator-activated receptor gamma was shown to be present in all three endocrine cell types studied (alpha, beta and delta cells) by immunohistochemistry. CONCLUSION/INTERPRETATION: We found that peroxisome proliferator-activated receptor gamma is highly expressed in human islet endocrine cells, both at the mRNA and protein levels. These results support the hypothesis of a direct influence of peroxisome proliferator-activated receptor gamma agonist on human pancreatic endocrine cells. PMID- 11043864 TI - Immunomagnetic purification of beta cells from rat islets of Langerhans. AB - AIMS/HYPOTHESIS: The aim of this study was to develop immunomagnetic purification by the Dynabead system to separate insulin-containing beta cells from a mixed rat islet cell population. Functional studies on insulin secretion and a test of the susceptibility of Dynabead-separated beta cells to DNA damage following cytokine exposure were carried out. METHODS: Dynabeads are uniform, paramagnetic particles coated with specific antibodies. Single rat islet cells were initially incubated with the beta-cell surface specific antibody (K14D10 mouse IgG) for 20-60 min. A suspension of Dynabeads coated with a secondary antibody (anti-mouse IgG) was added for a further 15 min, after which the Dynabead-coated cells were instantaneously pelleted by contact between the tube and a magnet (Dynal MPC). Immunocytochemistry was used to confirm that the Dynabead-coated cells contained insulin and to quantify the efficiency of the method. Dynabead-coated and non coated cells were stained for insulin and glucagon. RESULTS: Dynabead immunopurification yielded 95% pure insulin-containing beta cells, which released insulin in response to isobutylmethylxanthine and glucagon-like polypeptide 1. The insulin content of Dynabead-coated beta cells was significantly higher than that of non-coated cells. Successful separation was achieved using as few as 30 islets as starting material. Using the comet assay, we found that Dynabead-coated beta cells showed equal susceptibility to cytokine-induced DNA damage as non coated cells. CONCLUSION/INTERPRETATION: We conclude that Dynabead separation of beta cells is simple, rapid, applicable to large or small numbers of islets and can be used to study beta-cell specific function and responsiveness. PMID- 11043865 TI - Reduced capillary hydraulic conductivity in skeletal muscle and skin in Type I diabetes: a possible cause for reduced transcapillary fluid absorption during hypovolaemia. AB - AIMS/HYPOTHESIS: Patients with Type I (insulin-dependent) diabetes mellitus have a reduced transcapillary fluid absorption from skeletal muscle and skin and thus defective plasma volume regulation during hypovolaemia. Our aim was to find whether a defective capillary filtration coefficient or impaired transcapillary driving force are aetiologic factors for this reduction. METHODS: We investigated 11 diabetic patients (diabetes duration 6.9 +/- 1.1 years, age 26 +/- 1 years), without complications and 12 control subjects (26 +/- 1 years). Their capillary filtration coefficient was measured in the upper arm using a volumetric technique at rest and during lower body negative pressure (LBNP). We calculated the driving force for transcapillary fluid transfer. RESULTS: The increase in heart rate and the decrease in systolic blood pressure during lower body negative pressure were similar in diabetic and control subjects. The resting capillary filtration coefficient was decreased in the diabetic subjects, 0.033 +/- 0.003 vs 0.051 +/- 0.007 ml x 100 ml(-1) x min(-1) x mmHg(-1) (p < 0.05). During lower body negative pressure, the capillary filtration coefficient increased 35 % in both groups compared with resting capillary filtration coefficient and was still decreased in diabetes; 0.046 +/- 0.004 compared with 0.069 +/- 0.006 ml x 100ml(-1) x min(-1) x mmHg(-1) (p < 0.01). The established driving force during lower body negative pressure was 1.37 +/- 0.11 vs 1.30 +/- 0.15 mmHg (NS) in diabetic and control subjects, respectively. CONCLUSIONS/INTERPRETATION: Our study indicates that a reduced capillary filtration coefficient rather than defective regulation of transcapillary driving force, is the reason for the reduced transcapillary fluid absorption during hypovolaemic circulatory stress found in Type I diabetic patients. PMID- 11043866 TI - Effects of gestational diabetes on junctional adhesion molecules in human term placental vasculature. AB - AIMS/HYPOTHESIS: The aim of this study was to investigate whether gestational diabetes mellitus, which occurs in the microvascular remodelling phase of placental development, causes alterations in surface expression of tight and adherens junctional molecules involved in endothelial barrier function and angiogenesis. METHODS: Term placenta, delivered by elective Caesarian section, from normal pregnancy (n = 5) and those complicated by gestational diabetes (n = 5) were perfusion-fixed and analysed by indirect immunofluorescence and confocal scanning microscopy. Using systematic random sampling, the surface expression of endothelial junctional proteins and the relative incidences of immunostained vessels were compared between the two study groups. Total vessel lengths were measured by stereological techniques. RESULTS: The adherens junctional molecules, vascular-endothelial cadherin and beta-catenin, and the tight junctional molecules, occludin and zonula occludens-1 were localised to paracellular clefts in both study groups. The diabetic placentae showed pronounced reductions in the intensity of immunofluorescence and in the number of immuno-positive vessels. A corresponding statistically significant increase (from 19% to 56%) in the percentage of vessels showing junctional anti-phosphotyrosine immunoreactivity was found. The differences observed represented real changes in the absolute lengths of immunostained regions along the vessels. The stereological measurements failed to detect any statistically significant change in the combined length of fetal vessels in gestational diabetic placenta. CONCLUSION/INTERPRETATION: Our results suggest that even short duration diabetic insult, alters the surface expression of placental junctional proteins. This alteration could be mediated by the tyrosine-phosphorylation pathway. The changes suggest impaired barrier function rather than accelerated vascular growth. PMID- 11043867 TI - Identification of missense mutations in the hepatocyte nuclear factor-3beta gene in Japanese subjects with late-onset Type II diabetes mellitus. AB - AIMS/HYPOTHESIS: Hepatocyte nuclear factor (HNF)-3beta, a transcription factor expressed in pancreatic beta cells, is an upstream regulator of HNF-1alpha/MODY3, HNF-4alpha/MODY1 and IPF1/MODY5 genes. Our previous screening of MODY subjects showed that mutations in the HNF-3beta gene are not a common cause of this form of diabetes in the Japanese. We tested the hypothesis that mutations in the HNF 3beta gene cause late-onset Type II (non-insulin-dependent) diabetes mellitus in this population. METHODS: Genotyping of the polymorphic TCC repeat in the HNF 3beta gene was done in 112 Japanese subjects with Type II diabetes (age at diagnosis > 35 and family history of Type II diabetes among their second-degree relatives) and 96 Japanese control subjects. Furthermore, we screened 57 Type II diabetic patients for mutations of the HNF-3beta gene. Transactivation activity of variant HNF-3beta was investigated by transfection assay. RESULTS: The distribution of alleles of the TCC repeat was similar between diabetic and control groups. Mutation screening identified two missense mutations, A86T and G114E. Neither mutation was observed in 225 control subjects. The transactivation activity of G114E-HNF-3beta was similar to that of wild typeHNF-3beta. In contrast, the activity of A86T-HNF-3beta was statistically significantly reduced to 83-86 % of that of wild type. CONCLUSIONS/INTERPRETATION: The A86T mutation in the HNF-3beta gene might be involved in the development of late-onset Type II diabetes in a small group of Japanese people. PMID- 11043868 TI - The Gly972Arg variant in insulin receptor substrate-1 is not associated with birth weight in contemporary English children. The ALSPAC Study Team. Avon Longitudinal Study of Pregnancy and Childhood. PMID- 11043869 TI - R127W in HNF4alpha is a loss-of-function mutation causing maturity-onset diabetes of the young (MODY) in a UK Caucasian family. PMID- 11043870 TI - Report on a 1999 EASD Travel Fellowship for Young Scientists. European Association for the Study of Diabetes. PMID- 11043871 TI - Acute whiplash-associated disorders (WAD): the effects of early mobilization and prognostic factors in long-term symptomatology. AB - OBJECTIVE: To compare two different home exercise programmes for patients with acute whiplash-associated disorders (WAD). A further aim was to describe the initial prognostic variables related to self-reported pain at six months follow up. DESIGN: A randomized treatment study with a follow-up period of six months. SETTINGS: The study was undertaken in an orthopaedic clinic at a university hospital. SUBJECTS: A total of 59 symptomatic (neck pain, stiffness, etc.) patients with acute whiplash injury. INTERVENTIONS: Patients were randomized to a regular treatment group (RT group) and an additional-exercise treatment group (AT group). MAIN OUTCOME MEASURES: Pain Disability Index (PDI), Self-Efficacy Scale (SES), Coping Strategies Questionnaire (CSQ), neck range of motion (ROM), head posture, kinaesthetic sensibility, visual analogue scale (VAS). RESULTS: Patients given an additional exercise did not improve more than patients with regular treatment. Only one CSQ item, 'Ability to decrease pain', showed a significant difference between the groups in its pattern of change over time: the AT group had a significant increase between three and six months whilst values in the RT group decreased. Nonsymptomatic patients at six months follow-up were characterized by initially better self-efficacy, lower disability and significantly different patterns in the use of 'behavioural coping strategies' when compared with symptomatic patients. The nonsymptomatic patients also reported more frequent training than symptomatic patients, i.e. they complied better with the treatment regime. CONCLUSION: This home exercise programme, including training of neck and shoulder ROM, relaxation and general advice seems to be sufficient treatment for acute WAD patients when used on a daily basis. Additionally, patients reporting low self-efficacy and high disability levels may profit from more attention initially, as these psychological factors are significant predictors of pain at long-term follow-up. PMID- 11043872 TI - Clinical standards for inpatient specialist rehabilitation services in the UK. AB - OBJECTIVE: To develop a set of clinical standards for specialist inpatient rehabilitation services in the UK and to undertake a preliminary survey of consultants who provide those services. DESIGN: The proposed set of standards was developed by group consensus followed by an iterative consultation process. A postal survey was conducted on behalf of the British Society for Rehabilitation Medicine (BSRM) amongst its consultant members in the UK (n = 163), who were asked to assess their services in relation to these standards, and to comment on the standards themselves, their usefulness and applicability. RESULTS: The response rate was 61%, of which 81 respondents ran an inpatient rehabilitation service. Overall, the standards appeared to be acceptable to most, and mainly struck the right level, being attained by the majority of services. Specific suggestions were incorporated into the revised standards. Further work is required to establish agreed outcomes that are systematically measured and recorded: only half the respondents (50%) routinely recorded a standardized outcome measure, and only a quarter (26%) routinely reviewed patients to record long-term outcome. CONCLUSIONS: Clinical standards have been developed for specialist inpatient rehabilitation services in the UK. The BSRM proposes to adopt these standards for a test period of 2-3 years in the first instance. It is likely that they will require further refinement with time, and modification is required to adapt them to different subspecialities and settings. PMID- 11043873 TI - An epidemiological survey of the health needs of disabled people in a rural community. AB - OBJECTIVE: To describe service use and unmet needs of disabled people in a rural environment, given well-recognized difficulties in providing equitable services to a widely-spread population, availability of transport and the presence of discrepancies in wealth. PARTICIPANTS: Disabled people registered with a single rural general practice, identified by postal questionnaire. METHOD: All 3462 households were screened using the Office of Population, Censuses and Surveys (OPCS) disability screening questionnaire. An 86% response rate was achieved. Seventy-four people aged 16-65 and 69 people aged 66-75 were interviewed by a rehabilitation physician. Fifty-five people aged 76+ were interviewed. Disability was assessed using the OPCS scales of disability, Barthel Index, and Hospital Anxiety and Depression Scale. RESULTS: Thirty per cent (43) of those aged 16-75 received assistance for their personal activities of daily living (ADLs), and 98% (140) required assistance for extended ADLs (taken from the OPCS surveys criteria). There was no difference in either disability or dependency by age group. Fifty-three per cent of this group had domestic adaptations, 75% had disability aids. Those aged 16-75 had significantly fewer aids and adaptations, less home care, care management, respite, district nursing and chiropody services than people over 75. Equipment was provided by statutory services less frequently and fewer carers were salaried. The rehabilitation physician assessed them as needing more occupational therapy, physiotherapy and chiropody. Thirty per cent saw their GP monthly and 45% attended hospital. CONCLUSION: Unmet need was assessed as greater in the younger group. Elderly people are possibly more visible to service providers with better recognition of need. There is no evidence of a relationship between medical surveillance and identification of rehabilitation needs. Those with greater degrees of disability require more interlinked and organized services. PMID- 11043874 TI - Goal-setting in rehabilitation: report of a workshop to explore professionals' perceptions of goal-setting. AB - OBJECTIVE: To explore the views of therapists, nurses and doctors working in a variety of rehabilitation settings on the goal-setting process. METHODS: Sixteen rehabilitation staff from three different settings attended a goal-setting workshop in which the difficulties associated with goal-setting were described with their potential solutions. RESULTS: Five difficulties with goal-setting were identified. (1) Formal goal-setting was felt to be an activity unusual for many of the patients although intrinsic to the activity of many professional groups. (2) Goal-setting is often insensitive to people's roles in the community. (3) Goal-setting in the hospital environment does not transfer easily to the community. (4) External factors (e.g. staff turnover) over which staff felt they had little control were important in the success of a goal-setting programme. (5) Goals tend to be formulated and owned by the team, rather than the patient. CONCLUSIONS: Goal-setting is a very satisfactory activity for the team but to be as successful for the patient their needs must be acknowledged. People with mild disability and a short inpatient stay have different needs to those with acute onset severe permanent disability and those with chronic or progressive disability admitted from the community. Comparison of both process and outcome effects of different types of goal-setting is an area for future study. PMID- 11043875 TI - Balance function and fall-related efficacy in patients with newly operated hip fracture. AB - OBJECTIVE: To investigate the relation between fall-related efficacy in daily life activities and functional as well as instrumental tests of balance in patients with hip fracture. DESIGN: Analysis of different aspects of balance using the Falls Efficacy Scale, Swedish version FES(S), questions on fear of falling, Functional Reach (FR) and tests on a balance platform (Chattanooga). SUBJECTS: Fifty-five elderly inpatients (mean age 82.3) with newly operated hip fracture who were assessed during the last week in hospital before discharge. RESULTS: The results showed a significant relationship between the subjective ability measured with the FES(S) and the objectively measured balance in the Functional Reach test and also between fall-related efficacy measured with FES(S) and fear of falling. Very few significant correlations were found between the results from balance tests on the force platform and those obtained with FES(S) and FR. CONCLUSIONS: Both the Falls Efficacy Scale, Swedish version, and the Functional Reach have been shown to be useful in analysing balance function in elderly patients newly operated on for hip fracture. The Falls Efficacy Scale also indicates which of the daily activities the patient perceives as troublesome and thus require further training. PMID- 11043876 TI - Service needs of parents with motor or multiply disabled children in Dutch therapeutic toddler classes. AB - OBJECTIVE: To determine parents' service needs at the start of therapeutic toddler class treatment, to analyse determinants and to investigate received help after a period of 10 months. SUBJECTS: Parents with motor or multiply disabled children in therapeutic toddler classes. SETTING: Sixteen out of 17 Dutch rehabilitation centres. DESIGN: A sample of 84 parents, stratified according to toddler class size. MAIN OUTCOME MEASURES: A questionnaire of 33 items, divided into five categories, was designed to assess parents' needs and to evaluate the amount of received help. Several measures were used to assess child, parent and family characteristics. RESULTS: Based on factor analysis, five need subscales were distinguished which only partially corresponded to the initial categories. The initial category concerning needs for help in family functioning did not come out as a factor. Needs expressed most frequently belonged to the three subscales of needs for information concerning the child. The average level of needs generally did not differ for mothers and fathers across the subscales. However, we found large variation in needs for both mothers and fathers on each subscale. The most important, positively correlated determinant of parents' needs was social isolation of the family. After 10 months a considerable level of unmet needs was found. CONCLUSIONS: A systematic and periodic assessment of individual needs of parents as well as the services offered is recommended. Professionals of the therapeutic toddler classes should especially focus on providing parents with information on their child's disability and therapy, community services and parenting. PMID- 11043877 TI - A pilot study comparing the cognitive demand of walking for transfemoral amputees using the Intelligent Prosthesis with that using conventionally damped knees. AB - OBJECTIVE: To compare the cognitive demand of walking when using a conventional prosthesis with that using a microprocessor-controlled prosthesis. DESIGN: Ten unilateral transfemoral amputees wearing conventional pneumatic swing phase control (conventional prosthesis) prostheses walked on a treadmill which enforced a pattern of constantly varying speeds. The subjects simultaneously performed a simple or a complex distracting task. Following a period of accustomization, the subjects performed the same test wearing a prosthesis with microprocessor control of swing phase damping (the Intelligent Prosthesis). OUTCOME MEASURES: The three dimensional trajectory (sway) of a retroreflective marker attached to the forehead was measured by a video-based motion analysis system, and used as a measure of gait quality. The ratio of the sway for the complex task over the simple task (the 'automation index') was used as a measure of the degree of automation of gait. RESULTS: No significant differences were found in the automation index between the two devices. However, the total sway for the conventional prosthesis was significantly higher. Sway during the complex distracting task was significantly higher than during the simple task. CONCLUSIONS: The microprocessor-controlled prosthesis was not found to be less cognitively demanding than a conventional prosthesis. PMID- 11043878 TI - Obstetrical brachial plexus injuries: incidence, natural course and shoulder contracture. AB - The incidence of obstetric brachial plexus injury (OBPI) was investigated and the natural course of this disorder and the frequency of shoulder contracture described. Between 1988 and 1997 13,366 children with a gestational age of 30 weeks or more, were born at the Academic Medical Center, Amsterdam. Of these, 62 had an OBPI (4.6 per 1000). Complete neurological recovery occurred in 72.6% of cases. Half of them had a delayed recovery of more than three weeks (mean recovery time 6.2 +/- 3.1 months). Shoulder contracture occurred in at least one third of the children with delayed recovery and in at least two-thirds of the children with incomplete recovery. The incidence of OBPI in our hospital was found to be higher and to have a less favourable natural course than is usually reported in the literature. Contracture of the shoulder joint is frequently found even in infants with complete neurological recovery. PMID- 11043879 TI - Measuring the outcomes of day hospital attendance: a comparison of the Barthel Index and London Handicap Scale. AB - OBJECTIVE: To assess the ability of two scales to measure the effects of attendance at a geriatric day hospital. DESIGN: 'Before-and-after' measurements. SETTING: Day hospital serving a defined, urban, catchment area. SUBJECTS: One hundred and three consecutive new patients over a three-month period. INTERVENTION: Day hospital attendance for as long as the multidisciplinary team thought warranted. OUTCOME MEASURES: Barthel Index and London Handicap Scale. RESULTS: Seventy-six patients (74%) attended for broadly defined 'rehabilitation'. Measurements on the Barthel Index and London Handicap Scale were completed on 54 of these. Repeat measurements after discharge from the day hospital were achieved on 37 patients. Patients attended between two and 57 times, with a median of eight. Mean Barthel Index did not change over the period of attendance. There was a small improvement in mean handicap score. Eight patients were identified post hoc who attended for 10 or more sessions, and they experienced a large mean reduction in handicap. CONCLUSION: Overall, neither the Barthel Index nor the London Handicap Scale changed much during attendance at the day hospital. Generally very short lengths of attendance may have explained this. For patients with more prolonged attendance, who might be expected to change more, the London Handicap Scale proved more responsive than the Barthel Index. PMID- 11043880 TI - The Australian National Sub-acute and Non-acute Patient Casemix Classification (AN-SNAP): its application and value in a stroke rehabilitation programme. AB - OBJECTIVE: To investigate whether the Australian National Sub-acute and Non-acute Patient Casemix Classification (SNAP) and Functional Independence Measure and Functional Related Group (Version 2) (FIM-FRG2) casemix systems can be used to predict functional outcome, and reduce the variance of length of stay (LOS) of patients undergoing rehabilitation after strokes. DESIGN AND SETTING: The study comprised a retrospective analysis of the records of patients admitted to the Cedar Court Healthsouth Rehabilitation Hospital for rehabilitation after stroke. SUBJECTS: The sample included 547 patients (83.3% of those admitted with stroke during this period). Patient data were stratified for analysis into the five SNAP or nine FIM-FRG2 groups, on the basis of the admission FIM scores and age. MAIN OUTCOMES: The AN-SNAP classification accounted for a 30.7% reduction of the variance of LOS, and 44.2% of motor FIM, and the FIM-FRG2 accounts for 33.5% and 56.4% reduction respectively. Comparison of the Cedar Court with the national AN SNAP data showed differences in the LOS and functional outcomes of older, severely disabled patients. Intensive rehabilitation in selected patients of this type appears to have positive effects, albeit with a slightly longer period of inpatient rehabilitation. CONCLUSIONS: Casemix classifications can be powerful management tools. Although FIM-FRG2 accounts for more reduction in variance than SNAP, division into nine groups meant that some contained few subjects. This paper supports the introduction of AN-SNAP as the standard casemix tool for rehabilitation in Australia, which will hopefully lead to rational, adequate funding of the rehabilitation phase of care. PMID- 11043881 TI - Can young severely disabled stroke patients regain the ability to walk independently more than three months post stroke? AB - OBJECTIVE: To investigate the acquisition of ability to walk independently in young severely disabled stroke patients unable to walk three months after onset. SETTING: A regional rehabilitation unit providing post-acute neurorehabilitation mainly for patients aged 16-65 years. SUBJECTS AND METHODS: A retrospective analysis of records was undertaken for 152 stroke patients admitted consecutively over a three-year period. All had a combination of physical, cognitive and language impairments; male/female 98/54; median age 54 (IQR 47-60) years. MAIN OUTCOME MEASURES: Time taken from stroke onset to regain the ability to walk safely and independently at least 5 metres around the hospital bay. RESULTS: Seventy-five (49%) regained the ability to walk independently between 3 and 11 months post onset. Patients with cognitive impairments and neglect took longer to walk. Time from stroke onset to admission was longer in patients who did not walk (median 16.3 weeks), than in those who regained walking (median 12.7 weeks; p = 0.009). Independent positive associations were found between the time taken to walk and time from onset to referral for rehabilitation (p = 0.55; p < 0.001), and time from acceptance to admission (p = 0.30; p = 0.008). CONCLUSION: With intensive specialist input, significant numbers of severely disabled young stroke patients can regain independent walking many months after onset, though late walkers may follow a slower recovery course. These results argue for investment in appropriate rehabilitation for these patients. Waiting list delays may prolong rehabilitation and potentially compromise cost effectiveness. PMID- 11043882 TI - Are we missing urethral stricture after acquired brain injury? PMID- 11043883 TI - Sensitivity of Shah, Vanclay and Cooper's modified Barthel Index. PMID- 11043884 TI - Comorbid generalized anxiety disorder in primary social phobia: symptom severity, functional impairment, and treatment response. AB - As many as 50% of patients with a primary anxiety disorder may meet criteria for an additional anxiety disorder. However, there is insufficient research on the cooccurrence of the anxiety disorders, although investigations of this nature may facilitate our understanding of their cause, phenomenology, and treatment. The present study examined the occurrence of generalized anxiety disorder (GAD) among patients with social phobia (SP) compared with SP patients without GAD. Of 122 treatment-seeking patients meeting DSM-III-R criteria for SP, 29 (23.8%) also met criteria for an additional diagnosis of GAD. SP patients with comorbid GAD demonstrated greater severity on measures of social anxiety and avoidance, general anxiety, cognitive (but not somatic) symptoms of anxiety, depressed mood, functional impairment, and overall psychopathology. Group differences remained significant when comorbidity with other anxiety and mood disorders was controlled. The content of worry among the SP patients with GAD was not specific to social concerns and appeared similar to the reported content of worry in samples of patients with primary GAD. Nevertheless, SP patients with and without GAD responded similarly to cognitive-behavioral group therapy for social phobia. Implications for the understanding and treatment of comorbid SP and GAD are discussed. PMID- 11043886 TI - Recall and recognition memory for spider information. AB - Two experiments were devised to examine the effect of anxiety on memory for threatening information in those with spider phobia. Those with spider phobia, control participants, and those with other phobias were shown video clips of spiders. In the recognition task, participants were asked to state whether they remembered having seen specific video clips of spiders that had been presented to them previously. In the recall task, they were asked to remember and note as much detail as possible about each of a second series of video clips of spiders. Those with spider phobia were no different from the other two groups in their ability to recognize spider stimuli or in the amount of detail they were able to recall. PMID- 11043885 TI - A comparison of the efficacy of clonazepam and cognitive-behavioral group therapy for the treatment of social phobia. AB - There is a growing body of evidence that social phobia may be treated effectively by either pharmacologic or cognitive-behavioral interventions. but few studies have examined the relative benefits of these treatments. In this study, we examined the relative efficacy of pharmacotherapy with clonazepam and cognitive behavioral group therapy (CBGT) for treating social phobia. In addition, we examined potential predictors of differential treatment response. Outpatients meeting Diagnostic and Statistical Manual of Mental Disorders (3rd ed., revised) criteria for social phobia were randomly assigned to treatment. Clinician-rated and patient-rated symptom severity was examined at baseline and after 4, 8, and 12 weeks of treatment. All clinician-rated assessments were completed by individuals blind to treatment condition. Patients in both conditions improved significantly, and differences between treatment conditions were absent, except for greater improvement on clonazepam on several measures at the 12-week assessment. Symptom severity was negatively associated with treatment success for both methods of treatment, and additional predictors-sex, comorbidity with other anxiety or mood disorders, fear of anxiety symptoms, and dysfunctional attitudes failed to predict treatment outcome above and beyond severity measures. In summary, we found that patients randomized to clinical care with clonazepam or CBGT were equally likely to respond to acute treatment, and pretreatment measures of symptom severity provided no guidance for the selection of one treatment over another. PMID- 11043887 TI - Does a visual perceptual disturbance characterize trauma-related anxiety syndromes? AB - The i-test was developed to assess the visual-perceptual disturbances (VPDs) frequently reported by anxious patients. Persons with the disturbance report a specific abnormal illusion of movement when they maintain a fixed gaze at the i test stimulus. Base rates for positive responses to the i-test and for reports of a "recurrent specific memory" (RSM) of a fear experience were obtained in psychiatric outpatient (n = 301) and community (n = 128) samples. In each case, approximately one fifth of participants had a positive response to the i-test and one fifth of participants reported an RSM of fear. A positive response to the i test is observed in women more frequently than in men. Among psychiatric patients, approximately 90% of patients who report one symptom also report the other symptom; among community members, the concordance rate is approximately 33%. When psychiatric patients with both an abnormal illusion of movement response and an RSM of trauma are treated with eye movement desensitization, both symptoms are removed in 70% of cases; when these patients undergo some other form of treatment, both symptoms are removed in 30% of cases. These results indicate that the i-test is an effective way of identifying VPDs associated with psychopathologic conditions; the association between the abnormal illusion of movement and reports of recurrent specific memories of fear experiences suggests that the VPD may be a marker of traumatic stress syndromes. PMID- 11043888 TI - Generalized social phobia versus avoidant personality disorder: differences in psychopathology, personality traits, and social and occupational functioning. AB - Four groups of patients with social phobia (SP) were compared with regard to psychopathologic characteristics, personality traits, and social and occupational functioning. Fifteen persons with discrete social phobia without any personality disorder (DSP), 28 persons with generalized social phobia (GSP) without any personality disorder, 24 persons with GSP with a single diagnosis of avoidant personality disorder (APD), and 23 persons with GSP with more than one PD were included in the present study. APD had higher levels of social phobic avoidance, depressive symptoms, neuroticism, introversion, and social and occupational impairment as compared with GSP. DSP was found to be the least severe condition. OPD was the most impaired on nearly all variables. Logistic regression analyses revealed that introversion and depressive symptoms were able to predict correctly the presence or absence of an APD in 85% of those with social phobia. These findings are discussed in the light of the severity continuum hypothesis of social phobia and APD and recommendations for future research are given. PMID- 11043889 TI - Anxiety sensitivity, state and trait anxiety, and perception of change in sympathetic nervous system arousal. AB - The psychological models of panic disorder predict that persons with this condition may demonstrate enhanced acuity for somatic stimuli, but research to date has produced conflicting results. Most studies have investigated acuity for cardiovascular responses such as heart rate. which may represent an inadequate test of this hypothesis because they would be unlikely to detect persons who respond maximally to panic through other physiological systems. The present study investigated the detection of changes in pulse transit time (PTT), as a reliable and omnibus measure of sympathetic nervous system activity, with 36 healthy volunteers. We found that accurate perception of changes in PTT was consistently related to higher levels of trait anxiety and anxiety sensitivity, both of which are risk factors for the development of anxiety disorders in general. and panic disorder in particular. served. PMID- 11043890 TI - Structured and free-recall measures of worry themes: effect of order of presentation on worry report. AB - Structured instruments (SI) or free-recall instruments (FRI) can be used to examine worry themes. In the case where both types are use jointly, order of presentation may influence number and content of free-recall worries. Half of a large undergraduate sample (N = 609) received the SI before the FRI and the other half received them in the opposite order. Highly prevalent worries (i.e., common worries reported by more than 25% of the sample) were more numerous when the FRI was used first. However, moderately prevalent and rare worries (reported by 5% to 24% and 0% to 4% of the sample, respectively) were more numerous when the FRI was administered second. Results are discussed in terms of a possible priming effect produced by SI on moderately prevalent and rare free-recall worries. PMID- 11043891 TI - Searching for the stars. PMID- 11043892 TI - Talent identification and development in soccer. AB - In this review, we attempt to integrate the main research findings concerned with talent identification and development in soccer. Research approaches in anthropometry, physiology, psychology and sociology are considered and, where possible, integrated. Although some progress has been made in identifying correlates of playing success, it appears that no unique characteristics can be isolated with confidence. Both biological and behavioural scientists have indicated a strong genetic component in performance of sports such as soccer; nevertheless, the influence of systematic training and development programmes should not be underestimated. We conclude that the sport and exercise sciences have an important support role in the processes of identifying, monitoring and nurturing talented soccer players towards realizing their potential. PMID- 11043893 TI - Anthropometric and physiological predispositions for elite soccer. AB - This review is focused on anthropometric and physiological characteristics of soccer players with a view to establishing their roles within talent detection, identification and development programmes. Top-class soccer players have to adapt to the physical demands of the game, which are multifactorial. Players may not need to have an extraordinary capacity within any of the areas of physical performance but must possess a reasonably high level within all areas. This explains why there are marked individual differences in anthropometric and physiological characteristics among top players. Various measurements have been used to evaluate specific aspects of the physical performance of both youth and adult soccer players. The positional role of a player is related to his or her physiological capacity. Thus, midfield players and full-backs have the highest maximal oxygen intakes ( > 60 ml x kg(-1) x min(-1)) and perform best in intermittent exercise tests. On the other hand, midfield players tend to have the lowest muscle strength. Although these distinctions are evident in adult and elite youth players, their existence must be interpreted circumspectly in talent identification and development programmes. A range of relevant anthropometric and physiological factors can be considered which are subject to strong genetic influences (e.g. stature and maximal oxygen intake) or are largely environmentally determined and susceptible to training effects. Consequently, fitness profiling can generate a useful database against which talented groups may be compared. No single method allows for a representative assessment of a player's physical capabilities for soccer. We conclude that anthropometric and physiological criteria do have a role as part of a holistic monitoring of talented young players. PMID- 11043894 TI - Height, mass and skeletal maturity of elite Portuguese soccer players aged 11-16 years. AB - Height, mass and skeletal maturity (Fels method) were assessed in 135 elite youth soccer players aged 10.7-16.5 years (only two boys were < 11.0 years). Sample sizes, years of training and current weekly training volume by two-year age groups were: 11-12 years (n = 63), 2.6 +/- 1.0 years and 4.1 +/- 1.7 h; 13-14 years (n = 29), 3.1 +/- 1.6 years and 4.5 +/- 1.7 h; 15-16 years (n = 43), 4.7 +/ 2.4 years and 6.1 +/- 2.0 h. The oldest age group included members of the national youth team. Heights and masses were compared to US reference values, and skeletal age and chronological age were contrasted. The players were also classified as late, average ('on time') and early maturers on the basis of differences between skeletal and chronological age, with the average category including boys with skeletal ages within +/- 1 year of chronological age. The mean heights and masses of 11- to 12-year-old soccer players equalled the US reference values, while those of players aged 13-14 and 15-16 years were slightly above the reference values. The mean skeletal age approximated mean chronological age in players aged 11-12 years (12.4 +/- 1.3 and 12.3 +/- 0.5 years, respectively), while mean skeletal age was in advance of mean chronological age in the two older groups (14.3 +/- 1.2 and 13.6 +/- 0.7 years, respectively, in 13 to 14-year-olds; 16.7 +/- 1.0 and 15.8 +/- 0.4 years, respectively, in 15- to 16 year-olds). Seven boys in the oldest age group were already skeletally mature and were not included when calculating differences between skeletal and chronological age. The proportion of late maturing boys in this sample of elite soccer players decreased with increasing chronological age. Among 11- to 12-year-old players, the percentages of late and early maturing boys were equal at 21% (n = 13). Among 13- to 14-year-old players, the percentages of late and early maturing boys were 7% (n = 2) and 38% (n = 11) respectively, while among players aged 15-16 years the percentages of late and early maturing boys were 2% (n = 1) and 65% (n = 28) respectively. The results of this comparative analysis suggest that the sport of soccer systematically excludes late maturing boys and favours average and early maturing boys as chronological age and sport specialization increase. It is also possible that late maturing boys selectively drop-out of soccer as age and sport specialization increase. PMID- 11043895 TI - A multidisciplinary approach to talent identification in soccer. AB - The requirements for soccer play are multifactorial and distinguishing characteristics of elite players can be investigated using multivariate analysis. The aim of the present study was to apply a comprehensive test battery to young players with a view to distinguishing between elite and sub-elite groups on the basis of performance on test items. Thirty-one (16 elite, 15 sub-elite) young players matched for chronological age (15-16 years) and body size were studied. Test items included anthropometric (n = 15), physiological (n = 8), psychological (n = 3) and soccer-specific skills (n = 2) tests. Variables were split into separate groups according to somatotype, body composition, body size, speed, endurance, performance measures, technical skill, anticipation, anxiety and task and ego orientation for purposes of univariate and multivariate analysis of variance and stepwise discriminant function analysis. The most discriminating of the measures were agility, sprint time, ego orientation and anticipation skill. The elite players were also significantly leaner, possessed more aerobic power (9.0 +/- 1.7 vs 55.5 +/- 3.8 ml x kg(-1) x min(-1)) and were more tolerant of fatigue (P < 0.05). They were also better at dribbling the ball, but not shooting. We conclude that the test battery used may be useful in establishing baseline reference data for young players being selected onto specialized development programmes. PMID- 11043896 TI - Understanding and measuring coordination and control in kicking skills in soccer: implications for talent identification and skill acquisition. AB - In this review, we explore the role of motor control and biomechanics in developing an understanding of soccer skills using kicking as the main vehicle. The links between these sub-disciplines of sport science have not been well established in the past because of an emphasis on cognitive processes in traditional accounts of motor behaviour. We argue that a dynamical systems interpretation of the processes of coordination and control in movements with multiple degrees of freedom signals a new era in the relationship between the sub disciplines of motor control and biomechanics. Although research on coordination and control of soccer skills is currently sparse, there are indications that the relationship between motor control and biomechanics could form a significant component of scientific programmes in talent identification and skill development. Further interdisciplinary work is needed to enhance understanding of coordination and control of soccer skills. PMID- 11043897 TI - Psychological characteristics and talent identification in soccer. AB - I review research on psychological characteristics and sports performance and examine the literature on talent identification with particular reference to soccer to derive implications for the use of psychological variables in the talent identification and development process. Although the many cross-sectional studies of psychological characteristics and performance in all football codes conducted over the last 30 years have revealed no clear patterns, studies of both general inventories and specific variables are still being conducted. Reports on talent identification in all codes have increased in recent years, but most are descriptive in nature. In this review, I suggest that research on systematic expert observation has potential as a practical approach, but more studies of this type are needed. Considering the examination of specific psychological variables, only a solitary investigation of creativity in adolescents has shown promise. Further research on creativity and talent identification is required to replicate the positive results found in that study. In summarizing the research on psychological characteristics and talent identification, I conclude that cross sectional research on adults cannot be extrapolated for use in talent identification with adolescents. I propose that resources would be more effectively used in the provision of psychological skills training for adolescent soccer players, pending more sophisticated research on a wider range of psychological variables. It is recommended that longitudinal or quasi longitudinal research is essential to determine whether the same psychological variables are important for outstanding performance throughout the process of development and whether psychological variables measured during adolescence can predict outstanding performance in adulthood. PMID- 11043898 TI - The roles of talent, physical precocity and practice in the development of soccer expertise. AB - Here we consider the potential contributions of talent, physical precocity and deliberate practice in the development of soccer expertise. After presenting a working definition of 'talent', we examine how coaches perceive and select potential talent. Our findings suggest that much of what coaches see as early talent may be explained by physical precocity associated with a relative age advantage. Finally, as a test of the model of Deliberate Practice, we review the results of studies that assessed the progress of international, national and provincial players based on accumulated practice, amount of practice per week and relative importance and demands of various practice and everyday activities. A positive linear relationship was found between accumulated individual plus team practice and skill. Various practical suggestions can be made to improve talent detection and selection and to optimize career practice patterns in soccer. PMID- 11043899 TI - Perceptual skill in soccer: implications for talent identification and development. AB - In this review, key components of perceptual skill in soccer are identified and implications for talent identification and development highlighted. Skilled soccer players can recall and recognize patterns of play more effectively than their less skilled counterparts. This ability to encode, retrieve and recognize sport-specific information is due to complex and discriminating long-term memory structures and is crucial to anticipation in soccer. Similarly, experts use their knowledge of situational probabilities (i.e. expectations) to anticipate future events. They have a better than average idea of what is likely to happen given a particular set of circumstances. Also, proficiency-related differences in visual search strategy are observed. Skilled players use their superior knowledge to control the eye movement patterns necessary for seeking and picking up important sources of information. The nature of the task plays an important role in constraining the type of search used. Skilled soccer players use different search strategies when viewing the whole field (i.e. 11 vs 11 situations) compared with micro-states of the game (i.e. 1 vs 1, 3 vs 3 situations). Visual search behaviour also differs between defensive and offensive plays. These observations have implications for the development of perceptual training programmes and the identification of potential elite soccer players. PMID- 11043900 TI - Talent identification and women's soccer: an Australian experience. AB - This study used a quasi-applied research model to identify and develop potentially talented female soccer players. Athletes aged 15-19 years with a background in team ball sports or athletics were targeted for recruitment using advertisements and promotions through various media. Interested athletes attended a 2-day programme of testing, which included assessment of anthropometric, physiological and skill attributes. A combination of factors was used in the final selection of 17 athletes to take part in a 12-month talent development programme. A pre-season programme of five training sessions per week was conducted for 2 months. This programme focused on enabling the players to acquire the necessary ball and game skills to perform competitively in a short time. The squad competed as a team in the reserve grade competition of an Australian state league. At the conclusion of the 25-game season, 10 players were selected for zone teams with two players progressing to state team selection within 6 months. The project demonstrates that it is possible to select potential female soccer players based on anthropometric, physiological and skill attributes. Selection procedures could be enhanced through the development of objective assessment tools that measure tactical and technical competence. Programmes such as this can offer an additional avenue of player recruitment in support of existing procedures. PMID- 11043901 TI - The impact of elite labour migration on the identification, selection and development of European soccer players. AB - In this study, we examined sociocultural aspects of the identification, selection and development of elite soccer players as part of wider processes of globalization, particularly worker migration. Patterns of migration were identified among the 704 players who comprised the national squads of the 32 nations contesting the finals of the 1998 World Cup in France. An analysis of the migration patterns within and between the six Confederations into which member nations of FIFA are grouped established the European Federation (UEFA) as soccer's core economy. The study is subsequently focused on Europe and, in particular, upon the import strategies of clubs in the four most popular destination countries - England, Germany, Italy and Spain. It is argued that, in light of European Union deregulation of worker migration between member states and, in particular, the Bosman judgement, European soccer is being reshaped. The identification and selection of elite players are producing migrant patterns that are seen increasingly to impact upon indigenous player development and, potentially, the viability and success of national teams. We argue that, although these developments are contoured in part by global economic factors, economic accounts alone do not provide an adequate understanding of them. A series of interrelated economic, political, cultural and social factors is at work. We conclude with a brief outline of the policy implications of the analysis. PMID- 11043902 TI - Therapeutic potential of neurotrophic factors and neural stem cells against ischemic brain injury. AB - Development of neuronal and glial cells and their maintenance are under control of neurotrophic factors (NTFs). An exogenous administration of NTFs protects extremely sensitive brain tissue from ischemic damage. On the other hand, it is now known that neural stem cells are present in normal adult brain, and have a potential to compensate and recover neural functions that were lost due to ischemic stroke. These stem cells are also under control of NTFs to differentiate into a certain species of neural cells. Thus, the purpose of this review is to summarize the present understanding of the role of NTFs in normal and ischemic brain and the therapeutic potential of NTF protein itself or gene therapy, and then to summarize the role of NTFs in stem cell differentiation and a possible therapeutic potential with the neural stem cells against ischemic brain injury. PMID- 11043903 TI - Cortical death caused by striatal administration of AMPA and interleukin-1 is mediated by activation of cortical NMDA receptors. AB - Striatal coadministration of interleukin-1beta (IL-1beta) with alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionate (S-AMPA) in rats results in widespread cortical cell death not caused by either treatment alone. This cortical damage was unaffected by cortical infusion of the AMPA-receptor antagonist NBQX. Cortical infusion of an NMDA-receptor antagonist D-AP5 significantly inhibited (57%; P < 0.05) cortical death, but had no effect on the local striatal death. Thus, cortical neuronal death induced by striatal S-AMPA and human recombinant interleukin-1beta (hrIL-1beta) is mediated by activation of NMDA receptors in the cortex. The authors propose that IL-1beta actions on AMPA-receptor mediated cell death may involve the activation of polysynaptic pathways from the striatum to the cortex. PMID- 11043904 TI - Activation of specific neuronal circuits by corticotropin releasing hormone as indicated by c-fos expression and glucose metabolism. AB - The neuropeptide corticotropin releasing hormone (CRH) is the central nervous system (CNS) transducer of stressful stimuli. Endogenous CRH is released from neuronal terminals in several central nervous system regions-for example, amygdala and hypothalamus-during stress, and exogenous CRH administration mimics stress-related behaviors and hormonal patterns. However, whereas the role of endogenous CRH as a stress neuromodulator has been established, recent findings suggest that the peptide also functions to influence cognitive, emotional, and neuroimmune functions by modulating neuronal communication in a number of circuits. Although anatomic and pharmacologic approaches have provided evidence for this wider spectrum of CRH actions, the discrete regions and specific circuits activated by CRH have not been fully elucidated. In this article, the authors report on the use of two complementary methods to discern specific regions and cell groups activated by the administration of CRH. Glucose metabolism analysis provided quantitative measures of CRH-induced activation, but at a regional resolution; expression of the immediate early gene c-fos permitted a single cell resolution, but underestimated the neuroanatomic extent of CRH induced activation. Overlapping regions activated using both methods delineated discrete cortical, limbic. and motor pathways. Importantly, cell groups activated by CRH included those possessing either or both members of the CRH receptor family, suggesting that both receptors may mediate the effects of the endogenous ligand. In summary, CRH activates a broad but selective array of neuronal structures belonging to cortical, limbic, and motor circuits. These findings indicate that stress-related release of this peptide may contribute to a spectrum of important modulations of CNS function. PMID- 11043905 TI - Tolerance-Inducing dose of 3-nitropropionic acid modulates bcl-2 and bax balance in the rat brain: a potential mechanism of chemical preconditioning. AB - Many studies have reported ischemia protection using various preconditioning techniques, including single dose 3-nitropropionic acid (3-NPA), a mitochondrial toxin. However, the cellular signal transduction cascades resulting in ischemic tolerance and the mechanisms involved in neuronal survival in the tolerant state still remain unclear. The current study investigated the mRNA and protein expression of the antiapoptotic bcl-2 and the proapoptotic bax. two antagonistic members of the bcl-2 gene family, in response to a single dose of 3-NPA, to global cerebral ischemia-reperfusion. and to the combination of both 3-NPA pretreatment and subsequent global cerebral ischemia-reperfusion. Brain homogenates of adult Wistar rats (n = 25) were analyzed for bcl-2 and bax mRNA expression using a new highly sensitive and quantitative polymerase chain reaction (PCR) technique that allows real-time fluorescence measurements of the PCR product (LightCycler; Roche Diagnostics, Mannheim, Germany). Animals for mRNA analysis received 3-NPA (20 mg/kg, intraperitoneal; "chemical preconditioning") or vehicle (normal saline), and were either observed for 24 plus 3 hours or were subjected to 15 minutes of global cerebral ischemia 24 hours after the pretreatment and observed for 3 hours of reperfusion. Immunohistochemistry was applied to serial brain sections of additional rats (n = 68) to determine amount and localization of the respective Bcl-2 and Bax protein expression in various brain areas. One set of animals was injected with 3-NPA and observed for 3, 12, 24, and 96 hours; a second set was exposed to 15 minutes global cerebral ischemia, 3, 12, and 24 hours reperfusion; and a third set was pretreated with 3 NPA or saline 24 hours before the ischemic brain insult and observed for 96 hours of reperfusion. The authors found single dose 3-NPA treatment to be associated with an elevated bcl-2:bax ratio (increased bcl-2 expression, decreased bax expression), both on the transcriptional (mRNA) and the translational (protein) level. The differential influence of 3-NPA was maintained during early recovery from global cerebral ischemia (3 hours), when 3-NPA pretreated animals showed higher bcl-2 and lower bax mRNA levels compared with rats with saline treatment. Respective changes in protein expression were localized predominately in neurons vulnerable to ischemic damage. Compared with baseline, Bcl-2 protein was significantly higher in surviving neurons at 96 hours after the insult, whereas Bax protein remained unchanged. However, at this late time of postischemic recovery (96 hours), the protein expression pattern of surviving neurons was not different between animals with and without 3-NPA pretreatment. To the authors' knowledge, the current study is the first report on the differential expression of pro- and antiapoptotic genes after a single, nonlethal dose of 3-NPA. The current results suggest alterations in the balance between pro- and antiapoptotic proteins as a potential explanation for the reported protection provided by chemical preconditioning using 3-NPA in rats. PMID- 11043906 TI - Changes in mint1, a novel synaptic protein, after transient global ischemia in mouse hippocampus. AB - Mints (munc18-interacting proteins) are novel multimodular adapter proteins in membrane transport and organization. Mint1, a neuronal isoform, is involved in synaptic vesicle exocytosis. Its potential effects on development of ischemic damage to neurons have not yet been evaluated. The authors examined changes in mint1 and other synaptic proteins by immunohistochemistry after transient global ischemia in mouse hippocampus. In sham-ischemic mice, immunoreactivity for mint1 was rich in fibers projecting from the entorhinal cortex to the hippocampus and in the mossy fibers linking the granule cells of the dentate gyrus to CA3 pyramidal neurons. Munc18-1, a binding partner of mint1, was distributed uniformly throughout the hippocampus, and synaptophysin 2, a synaptic vesicle protein, was localized mainly in mossy fibers. After transient global ischemia, mint1 immunoreactivity in mossy fibers was dramatically decreased at 1 day of reperfusion but actually showed enhancement at 3 days. However, munc18-1 and synaptophysin 2 were substantially expressed in the same region throughout the reperfusion period. These findings suggest that mint1 participates in neuronal transmission along the excitatory pathway linking the entorhinal cortex to CA3 in the hippocampus. Because mint1 was transiently decreased in the mossy fiber projection after ischemia, functional impairment of neuronal transmission in the projection from the dentate gyrus to CA3 pyramidal neurons might be involved in delayed neuronal death. PMID- 11043907 TI - Characterization of cerebral white matter damage in preterm infants using 1H and 31P magnetic resonance spectroscopy. AB - The biochemical characteristics of white matter damage (WMD) in preterm infants were assessed using magnetic resonance spectroscopy (MRS). The authors hypothesized that preterm infants with WMD at term had a persisting cerebral lactic alkalosis and reduced N-acetyl aspartate (NAA)/ creatine plus phosphocreatine (Cr), similar to that previously documented in term infants weeks after perinatal hypoxiaischemia (HI). Thirty infants (gestational age 27.9 +/- 3.1 weeks, birth weight 1,122 +/- 445 g) were studied at postnatal age of 9.8 +/- 4.1 weeks (corrected age 40.3 +/- 3.9 weeks). Infants were grouped according to the presence or absence of WMD on magnetic resonance (MR) images. The peak area ratios of lactate/Cr, NAA/Cr, myo-inositol/Cr, and choline (Cho)/Cr were measured from an 8-cm3 voxel in the posterior periventricular white matter (WM) using proton MRS. Intracellular pH (pHi) was calculated using phosphorus MRS. Eighteen infants had normal WM on MR imaging; 12 had WMD. For infants with WMD, lactate/Cr and myo-inositol/Cr were related (P < 0.01); lactate/Cr and pHi were not (P = 0.8). In the WMD group, mean lactate/Cr and myo-inositol/Cr were higher (P < 0.001, P < 0.05, respectively) than the normal WM group. There was no difference in the NAA/Cr, Cho/Cr, or pHi between the two groups, although pHi was not measured in all infants. These findings suggest that WMD in the preterm infant at term has a different biochemical profile compared with the term infant after perinatal HI. PMID- 11043908 TI - Early detection of irreversible cerebral ischemia in the rat using dispersion of the magnetic resonance imaging relaxation time, T1rho. AB - The impact of brain imaging on the assessment of tissue status is likely to increase with the advent of treatment methods for acute cerebral ischemia. Multimodal magnetic resonance imaging (MRI) demonstrates potential for selecting stroke therapy patients by identifying the presence of acute ischemia, delineating the perfusion defect, and excluding hemorrhage. Yet, the identification of tissue subject to reversible or irreversible ischemia has proven to be difficult. Here, the authors show that T1 relaxation time in the rotating frame, so-called T1rho, serves as a sensitive MRI indicator of cerebral ischemia in the rat. The T1rho prolongs within minutes after a drop in the CBF of less than 22 mL 100 g(-1) min(-1). Dependence of T1rho on spin-lock amplitude, termed as T1rho dispersion, increases by approximately 20% on middle cerebral artery (MCA) occlusion, comparable with the magnitude of diffusion reduction. The T1rho dispersion change dynamically increases to be 38% +/- 10% by the first 60 minutes of ischemia in the brain region destined to develop infarction. Following reperfusion after 45 minutes of MCA occlusion, the tissue with elevated T1rho dispersion (yet normal diffusion) develops severe histologically verified neuronal damage; thus, the former parameter unveils an irreversible condition earlier than currently available MRI methods. The T1rho dispersion as a novel MRI index of cerebral ischemia may be useful in determination of the therapeutic window for acute ischemic stroke. PMID- 11043909 TI - Oxidative stress-associated impairment of proteasome activity during ischemia reperfusion injury. AB - Numerous studies indicate a role for oxidative stress in the neuronal degeneration and cell death that occur during ischemia-reperfusion injury. Recent data suggest that inhibition of the proteasome may be a means by which oxidative stress mediates neuronal cell death. In the current study, the authors demonstrate that there is a time-dependent decrease in proteasome activity, which is not associated with decreased expression of proteasome subunits, after cerebral ischemia-reperfusion injury. To determine the role of oxidative stress in mediating proteasome inhibition, ischemia-reperfusion studies were conducted in mice that either overexpressed the antioxidant enzyme glutathione peroxidase [GPX 1(+)], or were devoid of glutathione peroxidase activity (GPX -/-). After ischemia-reperfusion, GPX 1(+) mice displayed decreased infarct size, attenuated neurologic impairment, and reduced levels of proteasome inhibition compared with either GPX -/- or wild type mice. In addition, GPX 1(+) mice displayed lower levels of 4-hydroxynonenal-modified proteasome subunits after ischemia reperfusion injury. Together, these data indicate that proteasome inhibition occurs during cerebral ischemia-reperfusion injury and is mediated, at least in part, by oxidative stress. PMID- 11043910 TI - Secondary deterioration of apparent diffusion coefficient after 1-hour transient focal cerebral ischemia in rats. AB - Recent investigations on transient focal cerebral ischemia suggested recovery of energy metabolism during early reperfusion, but followed by secondary energy failure. As disturbances of energy metabolism are reflected by changes of the apparent diffusion coefficient (ADC) of water, the aim of the current study was to follow the dynamics of the ADC during 1 hour of middle cerebral artery occlusion (MCAO) and 10 hours of reperfusion. The right MCA was occluded in male Wistar rats inside the magnet using a remotely controlled thread occlusion model. Diffusion-, perfusion-, and T2-weighted images were performed repetitively, and ADC, perfusion, and T2 maps were calculated and normalized to the respective preischemic value. The lesion volume at each time point was defined by ADC < 80% of control. At the end of 1-hour MCAO the hemispheric lesion volume was 22.3 +/- 9.0%; it decreased to 6.4 +/- 5.7% in the first 2 hours of reperfusion (P < 0.01), but then increased again, and by the end of 10 hours of reperfusion reached 17.3 +/- 9.3%. The mean relative ADC in the end ischemic lesion volume significantly improved within 2 hours of reperfusion (from 65.7 +/- 1.2% to 90.1 +/- 6.7% of control), but later declined and decreased to 75.4 +/- 7.3% of control by the end of the experiment. Pixels with secondary deterioration of ADC showed a continuous increase of T2 value during the first 2 hours of reperfusion in spite of ADC improvement, indicating improving cytotoxic, but generation of vasogenic edema during early reperfusion. A significant decrease of the perfusion level was not observed during 10 hours of recirculation. The authors conclude that the improvement of ADC in the early phase of reperfusion may be followed by secondary deterioration that was not caused by delayed hypoperfusion. PMID- 11043911 TI - Effect of thrombolysis on the dynamics of infarct evolution after clot embolism of middle cerebral artery in mice. AB - Reversible focal ischemia may lead to delayed tissue injury despite primary restoration of blood flow and metabolism. The authors investigated whether such delayed changes also occur after thrombolytic treatment of thromboembolic stroke. Clot embolism of the middle cerebral artery (MCA) was produced in C57/B16J mice by intracarotid injection of heterologous clots. One hour after embolism, one group was treated with intracarotid infusion of rt-PA (10 mg/kg). The untreated control group received an equal amount of vehicle. Just before onset of treatment and after 1, 3. 6, and 24 hours, animals were frozen in situ and cerebral blood flow (CBF), cerebral protein synthesis (CPS), ATP content, and DNA fragmentations (TUNEL) were imaged on cryostat sections using double tracer autoradiography. bioluminescence, and immunohistochemical techniques, respectively. In untreated animals (n = 20), CPS was suppressed in approximately 68% of hemispheric transsection at 1 hour after embolization. The ATP depleted area was smaller (approximately 58%), but between 6 and 24 hours it merged with that of CPS suppression. TUNEL-positive neurons became visible between 6 and 24 hours exclusively in regions with ATP depletion. rt-PA-induced thrombolysis (n = 20) led to the gradual improvement of blood flow. At 24 hours. ATP depletion was fully reversed and the CPS suppression area declined to approximately 16% of hemispheric transsection. Despite progressive metabolic recovery, large numbers of neurons became TUNEL-positive and animals died between 24 and 48 hours. Thrombolysis after clot embolism restores metabolic activity including protein synthesis, but the therapeutic benefit is limited by secondary injury that requires additional treatment to improve final outcome. PMID- 11043912 TI - Quantitative assessment of longitudinal metabolic changes in vivo after traumatic brain injury in the adult rat using FDG-microPET. AB - With the advent and emerging importance of neurobiology and its relation to behavior, scientists desire the capability to apply noninvasive, quantitative imaging of neuronal activity to small rodents. To this end, the authors' laboratory has developed microPET, a high-resolution positron emission tomography (PET) scanner that is capable of performing in vivo molecular imaging at a resolution sufficient to resolve major structures in the rat brain. The authors report in this article that, in conjunction with 2-[18F]fluoro-2-deoxyglucose (FDG), microPET provides accurate rates of cerebral glucose metabolism (59.7 to 108.5 micromol/100 g x min) in conscious adult rats as validated by within subject autoradiographic measurements (59.5 to 136.2 micromol/100 g x min; r = 0.88; F[1,46] = 168.0; P < 0.001). By conducting repeated quantitative scanning, the authors demonstrate the sensitivity and accuracy of FDG-microPET to detect within-subject metabolic changes induced by traumatic brain injury. In addition, the authors report that longitudinal recovery from traumatic brain injury-induced metabolic depression, as measured by quantitative FDG-microPET, is significantly correlated (r = 0.65; P < 0.05) to recovery of behavioral dysfunction, as assessed by the Morris Water Maze performance of the same rats, after injury. This is the first study to demonstrate that FDG-microPET is quantitative, reproducible, and sensitive to metabolic changes, introducing a new approach to the longitudinal study of small animal models in neuroscience research. PMID- 11043913 TI - Human brain beta-hydroxybutyrate and lactate increase in fasting-induced ketosis. AB - Ketones are known to constitute an important fraction of fuel for consumption by the brain, with brain ketone content generally thought to be low. However, the recent observation of 1-mmol/L levels of brain beta-hydroxybutyrate (BHB) in children on the ketogenic diet suggests otherwise. The authors report the measurement of brain BHB and lactate in the occipital lobe of healthy adults using high field (4-T) magnetic resonance spectroscopy, measured in the nonfasted state and after 2- and 3-day fasting-induced ketosis. A 9-mL voxel located in the calcarine fissure was studied, detecting the BHB and lactate upfield resonances using a 1H homonuclear editing sequence. Plasma BHB levels also were measured. The mean brain BHB concentration increased from a nonfasted level of 0.05 +/- 0.05 to 0.60 +/- 0.26 mmol/L (after second day of fasting), increasing further to 0.98 +/- 0.16 mmol/L (after the third day of fasting). The mean nonfasted brain lactate was 0.69 +/- 0.17 mmol/L, increasing to 1.47 +/- 0.22 mmol/L after the third day. The plasma and brain BHB levels correlated well (r = 0.86) with a brain-plasma slope of 0.26. These data show that brain BHB rises significantly with 2- and 3-day fasting-induced ketosis. The lactate increase likely results from ketones displacing lactate oxidation without altering glucose phosphorylation and glycolysis. PMID- 11043914 TI - Does labeled alpha-methyl-L-tryptophan image ONLY blood-brain barrier transport of tryptophan? PMID- 11043915 TI - Gastric tonometry and intramucosal pH--theoretical principles and clinical application. AB - Gastric or intestinal luminal tonometry is a method for monitoring critically ill patients. It offers an index of the adequacy of aerobic metabolism in a tissue that is particularly sensitive to alterations in its perfusion and oxygenation: the gut mucosa. It is based on the measuring the increase in tissue CO2 production that accompanies anaerobic metabolism. The method simply consists of a balloon in the stomach, which measures intramucosal pCO2. From this measurement and from the arterial bicarbonate concentration gastric intramucosal pH (pHi) can be calculated, assuming that bicarbonate concentration in the gastric mucosal tissue is in equilibrium with systemic arterial bicarbonate. Despite possible clinical benefit from the measurement and the therapy of low pHi values in critically ill patients, the theoretical, experimental and pathophysiological implications for the monitoring of intramucosal acidosis in the gut are not yet fully understood. There are still some open methodological questions crucial for further clinical interpretation. PMID- 11043916 TI - Prevalence of various antiphospholipid antibodies in pregnant women. AB - Antiphospholipid antibodies (APAs) are characterized as a heterogeneous population of autoantibodies directed against different target antigens, predominantly anionic phospholipids or phospholipid-containing structures. The presence of APAs has been strongly associated with a variety of clinical disorders including adverse pregnancy complications such as spontaneous abortions, pregnancy-induced hypertension, preeclampsia and intrauterine growth retardation. The purpose of this study was to compare the prevalence of anticardiolipin antibodies (ACAs), which are routinely examined, with APAs directed against phosphatidylserine (APS), phosphatidylinositol (API), phosphatidylethanolamine (APE) and phosphatidylcholine (APC) in the sera of pregnant women. We examined 410 serum samples of pregnant women hospitalized in the department for pathological pregnancies. They underwent prenatal biochemical screening of fetal congenital abnormalities in the first and the second trimester of gravidity. Anticardiolipin IgG and IgM were measured using commercial ELISA kits (ImmuLisa Anti-Cardiolipin Antibody), whereas APS, APE, API and APC were determined by our modified ELISA kit. Among 410 pregnant women we found 21 patients (5.1%) positive for ACA IgG (>20 GPL) and 30 patients (7.3%) positive for ACA IgM (>10 MPL). It was found that 7.8% of pregnant women had at least one high-titer APA IgG and 9.8% high-titer APA IgM. One third of ACA IgG or IgM positive sera contained polyspecific autoantibodies reactive to at least two various phospholipids. In the group of IgG ACA positive women, 28.6% patients were positive for APS, 28.6% were positive or moderately positive for API, 23.8% for APC and 19% for APE. In the group of IgM ACA positive women, 33.3% were also positive for APS, 26.7% for APE, 26.7% for API and 23.3% for APC were present. IgG and IgM ACA negative patients exhibited a significantly lower incidence of other APA than the group of ACA positive pregnant women. It still remains to clarify if the routine examination of APA reacting with other anionic and zwitterionic antigens other than cardiolipin would improve the probability of identifying women liable to adverse pregnancy complications. PMID- 11043917 TI - Variable expression of hypercholesterolemia in Apolipoprotein E2* (Arg136 --> Cys) heterozygotes. AB - In the process of population screening for apo E gene polymorphism with the PCR and subsequent restriction analysis, we identified a female who demonstrated heterozygosity for an unusual restriction fragment caused by the loss of a CfoI restriction site. Sequence analysis of the apo E gene was performed and a carrier of the mutant allele with C --> T substitution at cDNA position 3817 was identified, which caused an Arg136 --> Cys change. The first-line relatives have been screened for this rare mutation with PCR and restriction analysis of PCR products. The complete lipoprotein parameters have been determined in the probands family. In the family, only one child had the same mutant allele as his mother had. The proband (7.49 mmol/l) with her siblings had hypercholesterolemia and a high body mass index (BMI 31.6 kg/m2). By contrast, her son had a normal lipid spectrum with normal BMI. We described the mutation apo E2* (Arg136 --> Cys) in a family with elevated lipid levels, but there was no confirmation of the connection between this mutation and type III hyperlipoproteinemia or hyperlipoproteinemia at all. In the case of this mutation, other factors (mainly genetic) are important for the development of lipid metabolism disorders. PMID- 11043918 TI - Selective antioxidant enzymes during ischemia/reperfusion in myocardial infarction. AB - The study of ischemia/reperfusion injury included 25 patients in the acute phase of myocardial infarction (19 perfused, 6 remained non-reperfused as evaluated according to the time course of creatine kinase and CK-MB isoenzyme activity) and a control group (21 blood donors). Plasma level of malondialdehyde was followed as a marker of oxidative stress. Shortly after reperfusion (within 90 min), a transient increase of malondialdehyde concentration was detected. The return to the baseline level was achieved 6 h after the onset of therapy. The activity of a free radical scavenger enzyme, plasma glutathione peroxidase (GPx), reached its maximum 90 min after the onset of treatment and returned to the initial value after 18 h. The specificity of the GPx response was confirmed by comparing with both non-reperfused patients and the control group, where no significant increase was detected. The erythrocyte Cu,Zn-superoxide dismutase (SOD) did not exhibit significant changes during the interval studied in perfused patients, probably due to the stability of erythrocyte metabolism. In non-reperfused patients, a decrease of SOD was found during prolonged hypoxia. These results help to elucidate the mechanisms of fast activation of plasma antioxidant system during the reperfusion after myocardial infarction. PMID- 11043919 TI - Baroreflex control of heart rate in young and adult salt hypertensive inbred Dahl rats. AB - Baroreflex control of heart rate was studied in inbred salt-sensitive (SS/Jr) and salt-resistant (SR/Jr) Dahl rats that were subjected to chronic dietary sodium chloride loading (for 4 weeks) either in youth or only in adulthood, i.e. from the age of 4 or 12 weeks. Using phenylephrine administration to pentobarbital anesthetized male rats we have demonstrated the decreased baroreflex sensitivity (lower slope for reflex bradycardia) in young prehypertensive SS/Jr rats fed a low-salt diet as compared to age-matched SR/Jr animals. High salt intake further suppressed baroreflex sensitivity in young SS/Jr but not in SR/Jr rats. Baroreflex sensitivity decreased with age in SR/Jr rats, whereas it increased in SS/Jr rats fed a low-salt diet. Thus at the age of 16 weeks baroreflex sensitivity was much higher in SS/Jr than in SR/Jr animals. High salt intake lowered baroreflex sensitivity even in adult SS/Jr rats without affecting it in adult SR/Jr rats. Nevertheless, baroreflex sensitivity was significantly lower in young SS/Jr rats with a severe salt hypertension than in adult ones with a moderate blood pressure elevation. It is concluded that the alterations of baroreflex sensitivity in young inbred SS/Jr rats (including the response to high salt intake) are similar to those described earlier for outbred salt-sensitive Dahl rats. We have, however, disclosed contrasting age-dependent changes of baroreflex sensitivity in both inbred substrains of Dahl rats. PMID- 11043920 TI - Ca-ATPase of human myometrium plasma membranes. AB - We determined and characterized the Mg2+-dependent, Ca2+-stimulated ATPase (Ca ATPase) activity in cell plasma membranes from the myometrium of pregnant women, and compared these characteristics to those of the active Ca2+-transport already demonstrated in this tissue. Similarly to the Ca2+-transport system, the Ca2+ ATPase is Mg2+-dependent, stimulated by calmodulin, and inhibited by vanadate. The Km for Ca2+ activation is 0.40 microM, very similar to that found for active calcium transport, i.e. 0.25 microM. Consequently, this Ca2+-ATPase can be responsible for the active calcium transport across the plasma membranes of smooth muscle cells. PMID- 11043921 TI - Effect of neonatal MSG treatment on day-night alkaline phosphatase activity in the rat duodenum. AB - The day-night variation of food intake and alkaline phosphatase (AP) activity was studied in the duodenum of rats neonatally treated with monosodium glutamate (MSG) and saline-treated (control) rats. The animals were kept under light-dark conditions (light phase from 09:00 h to 21:00 h) with free access to food. AP activity was cytophotometrically analyzed in the brush-border of enterocytes separated from the tip, middle and cryptal part of the villi every 6 h over a 24 hour period. In comparison with the controls, MSG-treated rats consumed about 40% less food during the dark period and their 24-hour food intake was thus significantly lowered (P<0.001). On the other hand, the nocturnal feeding habit showed a similar pattern: food consumption was high during the night (65% vs. 75%) and the lowest consumption was found during the light phase (35% vs. 25%) in MSG-treated and control rats, respectively. In agreement with the rhythm of food intake, the highest AP activity was observed during the dark phase and was lowest during the light phase in both groups of animals. These significant day-night variations showed nearly the same pattern in the enterocytes of all observed parts along the villus axis. In comparison with the controls, a permanent increase of AP activity was observed in neonatal MSG-treated rats. This increase was more expressive during the dark phase of the day in the cryptal (P<0.001) and middle part of the villus (P<0.01). From the viewpoint of feeding, this enzyme in MSG-treated rats was enhanced in an inverse relation to the amount of food eaten i.e. despite sustained hypophagia the mean AP activity in the enterocytes along the villus axis was higher than in the control animals during all investigated periods. The present results suggest that the increased AP activity in MSG treated rats is probably not a consequence of actual day-night eating perturbations but could be a component of a more general effect of MSG. This information contributes to better understanding of the function of intestinal AP and its relation to day-night feeding changes especially in connection with the MSG syndrome. PMID- 11043922 TI - The influence of insulin on the in vitro development of mouse and bovine embryos. AB - To further investigate the role of insulin during preimplantation embryo development, we compared the effects of insulin on the development of mouse and bovine preimplantation embryos and on cell proliferation during culture in vitro in simplex media. The influence of insulin on the development of mouse zygotes was determined during cultivation in mSOF medium, alone or supplemented with glucose. Similarly, the effects of insulin on the bovine preimplantation embryo development were studied in mSOF medium. The addition of insulin into mSOF medium enhanced significantly the number of cells per mouse blastocyst. Moreover, when mSOF medium was supplemented with insulin and 0.2 mmol x l(-1) glucose, the percentage of hatched blastocysts and the mean cell number of mouse blastocysts were significantly higher. Insulin had no significant effect on the development of bovine embryos, produced by in vitro fertilization of in vitro matured oocytes. Neither the rates of developing embryos nor the mean number of cells in blastocysts were different in comparison with control embryos. Our results suggest that the in vitro development of mouse embryos could be enhanced by the addition of insulin to the culture medium and is further improved by the addition of glucose. In contrast to this our results indicate that insulin has no detectable beneficial effect on the preimplantation development of bovine embryos in mSOF medium. PMID- 11043923 TI - Effects of treatment with interleukin-1 receptor antagonist on endogenous interleukin-1 levels in normal and irradiated mice. AB - The in vivo effects of recombinant human interleukin-1 receptor antagonist (rhIL 1Ra) administration on endogenous IL-1 levels in the circulation and conditioned media (CM) from different immunohematopoietic organ/tissues were studied in CBA mice under steady state and postirradiation conditions. In normal mice, constitutive IL-1 levels were demonstrated in the plasma, CM of peritoneal exudate cells and full-thickness skin explants with low or undetectable levels in CM of splenic and bone marrow cell suspensions. In irradiated mice (2 Gy, X rays) on day 3 post exposure a significant increase of IL-1 levels was seen in the circulation and CM of peritoneal exudate cells, with no significantly different levels in postirradiation bone marrow, spleen and skin. After rhIL-1Ra treatment of the animals (2 x 50 microg/mouse, i.p.), significantly elevated IL-1 levels were observed in the skin and CM of peritoneal exudate cells in normal mice, whereas slightly increased levels were detected in CM of splenic cells. The rhIL 1Ra administration in irradiated mice led to decreased IL-1 concentrations in the circulation, and CM of peritoneal exudate cells and skin. The results pointed out the importance of IL-1 secretion and receptor expression in the maintenance of homeostasis in steady state, as well as during recovery after irradiation. Modulatory effects of IL-1Ra on IL-1 production were dependent on basic endogenous IL-1 concentration. PMID- 11043924 TI - Effects of vitamin E and prostaglandin E2 on expression of CREB1 and CREB2 proteins by human T lymphocytes. AB - Both prostaglandins (PGs) and vitamin E are known to deeply affect immune responses. It is shown here that they both influence T cell-mediated immunity through reciprocal interference on the expression of cyclic-AMP responsive element binding (CREB) family proteins. CREB1 protein of human T lymphocytes was significantly modulated by a brief treatment of 5 to 10 min with PGE2. On the contrary, vitamin E appeared to be ineffective on the CREB1 behavior, while it abolished the PGE2-induced modulation of this protein. The CREB2 protein expression was also affected by PGE2 treatment, but a longer period of incubation (>20 min) was needed to observe these changes. Vitamin E showed a strong enhancing effect on CREB2 that was partially reversed by the subsequent treatment with PGE2. Our results support the idea that there is reciprocal interference between PGE2 and vitamin E on PGE2-induced signals in T lymphocytes. These data are in agreement with the reports concerning different cell systems and experimental conditions. PMID- 11043925 TI - Discharge properties of neurons in subdivisions of the medial geniculate body of the guinea pig. AB - The activity of 194 neurons was recorded in three subdivisions of the medial geniculate body (74 neurons in the ventral, 62 in the medial and 44 neurons in the dorsal subdivision, i.e. vMGB, mMGB and dMGB) of guinea pigs anesthetized with ketamine-xylazine. The discharge properties of neurons were evaluated by means of peristimulus time histograms (PSTHs), interval histograms (INTHs) and auto-correlograms (ACGs). In the whole MGB, the most frequent PSTH responses to pure tone stimuli were onset (43%) or chopper (32%). The onset responses were mostly present in the vMGB, whereas chopper responses dominated in the dMGB. In the whole MGB Poisson-like and bimodal INTHs were found in 46% and 40% of neurons, respectively. The mMGB revealed fewer bimodal and more symmetrical types of INTH. In the whole MGB, 60% of units were found to have ACGs typical for short bursts (<100 ms), 23% for long bursts (>100 ms) and 15% of units fired without bursts. Neurons in the vMGB were characterized by short bursting, whereas those in the mMGB and dMGB expressed more activity in the long bursts. The results demonstrate that the type of information processing in the vMGB, which belongs to the "primary" auditory system, is different from that in two other subdivisions of the MGB. PMID- 11043926 TI - Effects of L-arginine on prevention and treatment of lithium-pilocarpine-induced status epilepticus. AB - The effects of various doses of L-arginine, a nitric oxide substrate, on lithium pilocarpine-induced seizures were studied in rats. Rats were implanted with chronic, stainless steel screw electrodes epidurally for electrocortical recordings. A control group received 3 mEq/kg LiCl (i.p.) and 24 h later 45 mg/kg pilocarpine HCl (i.p.). Two different experimental procedures were followed: (1) L-arginine was applied in doses of 100 mg/kg, 300 mg/kg or 500 mg/kg (i.p.), 30 min before pilocarpine injection; (2) 300 mg/kg, 500 mg/kg or 1000 mg/kg (i.p.) L arginine was injected either 5 min or 30 min after the onset of status epilepticus (SE). L-arginine (300 mg/kg) injected 30 min before pilocarpine significantly reduced the percentage of SE, but did not change the latency to SE or 24-hour survival. These parameters were not significantly affected by the 100 mg/kg or 500 mg/kg dose of L-arginine. On the other hand, no dose of L-arginine that was applied after SE had begun, had any significant influence on the seizures. We concluded that L-arginine may prevent seizure activity in some but not all doses, and does not have any effect on the ongoing seizure activity. PMID- 11043927 TI - Kainic acid lesions to the lateral tegmental field of medulla: effects on cough, expiration and aspiration reflexes in anesthetized cats. AB - We have tested the hypothesis that neurons of both the ventral reticular nucleus and the adjacent parts of the lateral tegmental field (LTF) may be important for the production of motor programs associated with cough, expiration and aspiration reflexes. Our studies were conducted on non-decerebrate, spontaneously breathing cats under pentobarbitone anesthesia. Dysfunction of the medullary LTF region above the obex, produced by uni- or bilateral injections of kainic acid (a neurotoxin), regularly abolished the cough reflex evoked by mechanical stimulation of both the tracheobronchial and laryngeal regions and in most cases also the expiration reflex induced from the glottal area. However, some electrical activity still occurred in the neurogram of the recurrent laryngeal nerve during probing the laryngeal and glottal regions. Interestingly, the aspiration reflex elicited from the nasopharynx regularly persisted, although with lower intensity after the LTF lesion. Nevertheless, successive midcollicular decerebration performed in four cats also abolished the aspiration reflex. These experiments demonstrate the importance of medullary LTF neurons for the normal occurrence of cough and expiration reflexes. One possible explanation for the elimination of these expulsive processes is that the blockade of the LTF neurons may remove an important source of a facilitatory input to the brainstem circuitries that mediate cough and expiration reflexes. In addition, the potential importance of the mesencephalic reticular formation for the occurrence of the aspiration reflex and the role of the LTF in modulating both the eupnoeic breathing and the blood pressure are also discussed. PMID- 11043928 TI - Correlation of carnitine levels to methionine and lysine intake. AB - Plasma carnitine levels were measured in two alternative nutrition groups--strict vegetarians (vegans) and lactoovovegetarians (vegetarians consuming limited amounts of animal products such as milk products and eggs). The results were compared to an average sample of probands on mixed nutrition (omnivores). Carnitine levels were correlated with the intake of essential amino acids, methionine and lysine (as substrates of its endogenous synthesis), since the intake of carnitine in food is negligible in the alternative nutrition groups (the highest carnitine content is in meat, lower is in milk products, while fruit, cereals and vegetables contain low or no carnitine at all). An average carnitine level in vegans was significantly reduced with hypocarnitinemia present in 52.9% of probands. Similarly, the intake of methionine and lysine was significantly lower in this group due to the exclusive consumption of plant proteins with reduced content of these amino acids. Carnitine level in lactoovovegetarians was also significantly reduced, but the incidence of values below 30 micromol/l was lower than in vegans representing 17.8% vs. 3.3% in omnivores. Intake of methionine and lysine was also significantly reduced in this group, but still higher compared to vegans (73% of protein intake covered by plant proteins). Significant positive correlation of carnitine levels with methionine and lysine intake in alternative nutrition groups indicates that a significant portion of carnitine requirement is covered by endogenous synthesis. Approximately two thirds of carnitine requirement in omnivores comes from exogenous sources. The results demonstrate the risks of alternative nutrition with respect to the intake of essential amino acids, methionine and lysine, and with respect to the intake and biosynthesis of carnitine. PMID- 11043929 TI - Minireview: analysis of rape seed napin structure and potential roles of the storage protein. AB - Structural and functional data on 2S albumins and particularly rape seed napins are reviewed and, based on the coordinates of the three-dimensional structure of napin-like albumin BnIb, are used to model different rape napins. Surprisingly, the modeled napins, despite great sequence homology, differ in tertiary arrangements of the polypeptide chains. It is proposed that these differences in 3D structures of the analyzed rape napins may reflect their functions, which may cover many other potential beneficial purposes besides simple storage. PMID- 11043930 TI - Perturbation of the intermolecular contact regions (molecular surface) of hemoglobin S by intramolecular low-O2-affinity-inducing central cavity cross bridges. AB - The general assumption among researchers on hemoglobin is that the intramolecular central cavity cross-bridging of Hb does not result in any generalized perturbations at the protein surface. A corollary of this is that central cavity cross-bridges are unlikely to influence the polymerization of deoxy HbS, since polymerization is a protein surface phenomenon involving the participation of multiple protein surface amino acid residues. In an attempt to evaluate this experimentally, we have introduced two low-O2-affinity-inducing central cavity cross-bridges into HbS, beta(beta)-sebacyl [between the two Lys-82(beta) residues] and alpha(alpha)-fumaryl [between the two Lys-99(alpha) residues], and investigated their influence on the polymerization of the deoxy protein. The O2 affinities of the cross-bridged HbS exhibited sensitivity toward the buffer ions and pH in a cross-link-specific fashion. The modulation of the O2 affinity of these cross-bridged HbS in the presence of allosteric effectors, DPG and L-35, is also very distinct, reflecting the differences in the conformational features these two cross-bridges induce within the central cavity at the respective effector-binding domains. In addition, the alpha(alpha)-fumaryl cross bridge inhibited the polymerization, reflecting the perturbation of the microenvironment of one or more intermolecular contact residues, protein surface residues, as a consequence of the central cavity cross-bridge. On the other hand, the beta(beta) sebacyl cross-bridge exerted a slight potentiating effect on the polymerization of HbS. This reflects the fact that the perturbations at the protein surface are limited and favor polymerization. The results presented demonstrate that the structural changes induced by the central cavity cross-bridges are very specific and not simply restricted to the sites of modification, but are propagated to distant sites/domains, both within and outside the central cavity. It is conceivable that other surface regions that are not involved in the polymerization could also experience similar structural/conformational consequences. These results should be taken into consideration in designing intramolecularly cross-bridged asymmetric hybrid HbS for mapping the contribution of the intermolecular contact residues in the cis and trans dimers of deoxy HbS during polymerization. PMID- 11043931 TI - Prediction of membrane protein types based on the hydrophobic index of amino acids. AB - A new algorithm to predict the types of membrane proteins is proposed. Besides the amino acid composition of the query protein, the information within the amino acid sequence is taken into account. A formulation of the autocorrelation functions based on the hydrophobicity index of the 20 amino acids is adopted. The overall predictive accuracy is remarkably increased for the database of 2054 membrane proteins studied here. An improvement of about 13% in the resubstitution test and 8% in the jackknife test is achieved compared with those of algorithms based merely on the amino acid composition. Consequently, overall predictive accuracy is as high as 94% and 82% for the resubstitution and jackknife tests, respectively, for the prediction of the five types. Since the proposed algorithm is based on more parameters than those in the amino acid composition approach, the predictive accuracy would be further increased for a larger and more class balanced database. The present algorithm should be useful in the determination of the types and functions of new membrane proteins. The computer program is available on request. PMID- 11043932 TI - Analysis of the disulfide bonding pattern between domain sequences of recombinant prochymosin solubilized from inclusion bodies. AB - Prochymosin contains three disulfide bonds linking Cys45 to Cys50, Cys206 to Cys210, and Cys250 to Cys283. To analyze the disulfide bonding pattern between domain sequences in the recombinant prochymosin molecule solubilized from inclusion bodies by 8 M urea (designated as solubilized prochymosin), a simple peptide mapping method was established. This process consists of thiol alkylation, cleavage with cyanogen bromide, diagonal electrophoresis on polyacrylamide gel, and N-terminal sequencing. By using this procedure it was found that Cys45 and Cys50 located in the N-terminal domain are not mispaired with the cysteine residues, located in the C-terminal domain, in the solubilized wild-type prochymosin and its mutants. This result implies that Cys45 and Cys50, the partners of a native disulfide, are restricted in some ordered structures existing in inclusion bodies and remaining after solubilization. These native structural elements act as folding nuclei to initiate and facilitate correct refolding. The strategy of preserving the native-like structures including native disulfide in the solubilized inclusion bodies to enhance renaturation efficiency may be applicable to other recombinant proteins. PMID- 11043933 TI - The two slow refolding processes of creatine kinase are catalyzed by cyclophilin. AB - A burst phase occurs in the refolding kinetics of guanidine-denatured creatine kinase due to formation of an intermediate within the mixing dead time, with further refolding to the native state after the burst phase along a path following biphasic kinetics. In the presence of cyclophilin, the refolding rates of the two slow processes are accelerated and the values are proportional to the cyclophilin concentration. The activity of cyclophilin in accelerating the slow refolding processes of creatine kinase is totally inhibited by cyclosporin A, indicating that the cis-trans isomerization of the peptidyl-prolyl bonds is involved in the two slow refolding processes. PMID- 11043934 TI - Molecular modeling of the interactions of trichosanthin with four substrate analogs. AB - Trichosanthin (TCS) is a ribosome-inactivating protein (RIP) that possesses N glycosidase activity. It inactivates ribosomes and arrests protein synthesis by removing a specific adenine from 28S rRNA. A molecular dynamics simulated annealing method was applied to study the binding modes of TCS with substrate analogs, three oligonucleotides GAG, GAGA, and CGAGAG, based on the crystal structures of the stable complexes of TCS with NADPH and with the reaction product adenine. A water molecule proposed to be responsible for hydrolyzing the N-glycosidic bond was included in the model. All the oligoribonucleotides can dock into the active cleft of TCS without unfavorable contacts. The interaction energies between TCS and the three oligonucleotides were calculated. The interactions of TCS with NADH were also studied by a molecular dynamics simulated annealing method. The interaction energy between NADH and TCS was compared with that between NADPH and TCS, showing that the lack of 2'-phosphate group leads to an energy rise of 20 kcal/mol. PMID- 11043935 TI - Structure and heterogeneity of the one- and two-disulfide folding intermediates of tick anticoagulant peptide. AB - Tick anticoagulant peptide (TAP) is a factor Xa-specific inhibitor and is structurally homologous to bovine pancreatic trypsin inhibitor (BPTI). The fully reduced TAP refolds spontaneously to form the native structure under a wide variation of redox buffers. The folding intermediates of TAP consist of at least 22 fractions of one-disulfide, two-disulfide, and three-disulfide scrambled isomers. Three species of well-populated one- and two-disulfide intermediates were isolated and structurally characterized. The predominant one-disulfide species contains TAP-(Cys33-Cys55). Two major two-disulfide isomers were TAP (Cys33-Cys55, Cys15-Cys39) and TAP-(Cys33-Cys55, Cys5-Cys39). Both Cys33-Cys55 and Cys15-Cys39 are native disulfides of TAP. These three species are structural counterparts of BPTI-(Cys30-Cys51), BPTI-(Cys30-Cys51, Cys14-Cys38), and BPTI (Cys30-Cys51,Cys5-Cys38), which have been shown to be the major intermediates of BPTI folding. In addition, time-course-trapped folding intermediates of TAP, consisting of about 47% one-disulfide species and 30% two-disulfide species, were collectively digested with thermolysin, and fragmented peptides were analyzed by Edman sequencing and mass spectrometry in order to characterize the disulfide containing peptides. Among the 15 possible single-disulfide pairings of TAP, 10 (2 native and 8 nonnative) were found as structural components of its one- and two-disulfide folding intermediates. The results demonstrate that the major folding intermediates of TAP bear structural homology to those of BPTI. However, the folding pathway of TAP differs from that of BPTI by (a) a higher degree of heterogeneity of one- and two-disulfide intermediates and (b) the presence of three-disulfide scrambled isomers as folding intermediates. Mechanism(s) that may account for these diversities are proposed and discussed. PMID- 11043936 TI - Dissociation of human alphaB-crystallin aggregates by thiocyanate is structurally and functionally reversible. AB - Conformational modifications and changes in the aggregation state of human alphaB crystallin were investigated at different concentrations of SDS, KBr, urea, and NH4SCN and at different temperatures. Intrinsic fluorescence measurements indicated complete and reversible unfolding of the protein at 2 M NH4SCN, whereas the concentration of urea required for complete and irreversible unfolding was 6 M. Gel permeation chromatography indicated almost complete dissociation of the micelle-like aggregate of alphaB-crystallin in 2 M NH4SCN, but only partial dissociation into large-sized aggregates in 6 M urea. Thiocyanate-treated alphaB crystallin recovered its chaperone-like activity upon dilution of the dissociating agent, whereas the urea-treated protein did not. PMID- 11043937 TI - Probing the roles of the only universally conserved leucine residue (Leu122) in the oligomerization and chaperone-like activity of Mycobacterium tuberculosis small heat shock protein Hsp16.3. AB - To understand the role of the only universally conserved hydrophobic residue among all the members of the sHsp family, this extremely well conserved Leu122 residue in Hsp16.3 was replaced by valine, alanine, asparigine, or aspartate residues. Only very small amounts of the L122D and L122N mutant Hsp16.3 proteins were expressed in the transformed E. coli; however, both the L122V and L122A were readily expressed. The L122V and L122A mutant proteins had similar oligomeric structures to the wild-type protein at room temperature. Examination of the L122A mutant protein by native pore gradient PAGE and CD spectroscopy, however, revealed a smaller oligomeric size and different secondary structure at 37 degrees C. Both L122V and L122A mutant proteins exhibited significantly lowered chaperone activities. Observations reported here suggest a very important role of this only universally conserved Leu residue in both the formation of specific oligomeric structures and the molecular chaperone activities of Hsp16.3. PMID- 11043938 TI - Thrombin inhibitor design: X-ray and solution studies provide a novel P1 determinant. AB - The crystal structures of proflavin and 6-fluorotryptamine thrombin have been completed showing binding of both ligands at the active site S1 pocket. The structure of proflavin:thrombin was confirmatory, while the structure of 6 fluorotryptamine indicated a novel binding mode at the thrombin active site. Furthermore, speculation that the sodium atom identified in an extended solvent channel beneath the S pocket may play a role in binding of these ligands was investigated by direct proflavin titrations as well as chromogenic activity measurements as a function of sodium concentration at constant ionic strength. These results suggested a linkage between the sodium site and the S1 pocket. This observation could be due to a simple ionic interaction between Asp189 and the sodium ion or a more complicated structural rearrangement of the thrombin S1 pocket. Finally, the unique binding mode of 6-fluorotryptamine provides ideas toward the design of a neutrally charged thrombin inhibitor. PMID- 11043939 TI - Baculovirus infection blocks the progression of fat body degradation during metamorphosis in Bombyx mori. AB - The effect of baculovirus infection into silkworm pupa particularly on programmed fat body degradation during metamorphosis was investigated. Pupal fat body degradation did not occur following infection with Bombyx mori nucleopolyhedrovirus (BmNPV). There were no histolytic differences between the fat body tissues of mock and BmNPV infected papae until 48 h postinfection (p.i.). Between 48 and 72 h p.i., significant differences were observed. In order to determine whether the histolysis of fat body was due to apoptosis, genomic DNAs were purified at various p.i. times and analyzed. Rapid genomic DNA fragmentation was observed at 24 and 48 h after pupation in both mock and BmNPV infected pupae. However, BmNPV infection clearly inhibited DNA fragmentation and ladder formation at 72 h and later times p.i. Furthermore, pulse-labeling analysis showed that BmNPV infection restored protein synthesis in fat body cells. These results suggested that fat body degradation during pupal-adult development was due to apoptosis, and that BmNPV was able to evoke a cellular response in these cells. PMID- 11043940 TI - Prevalence of equine herpesvirus type 1 latency detected by polymerase chain reaction. AB - In this study, an improved polymerase chain reaction (PCR) was used for detection of DNA of latent EHV-1 strains from several sources. Three pairs of oligonucleotide primers spanning fragments of 333 bp, 226 bp and 268 bp of the thymidine kinase (tk) gene, and one primer pair spanning 225 bp of the glycoprotein C (gC) gene were used in specific amplifications. Primers for EHV-4 PCR were also designed. Restriction digests with TaqI confirmed the identity of tk PCR fragments from EHV-1. The sensitivity to detect PCR products was further improved by visualisation in silver-stained acrylamide gels. PCR assays were applied to 267 samples including pools of tissue, peripheral blood leukocytes (PBL) and nasal swabs of archived, farms and abattoir specimens from a total of 116 animals. The EHV-1 DNA was found in 88% of the analysed samples. The prevalence of the EHV-1 latent or persistent form in adult horses was similar to others reports but found higher than previously described in foetuses and young foals. EHV-4 latency was not detected in the Brazilian studied specimens. PMID- 11043941 TI - Comparative sequence analysis identified mutations outside the NSP4 cytotoxic domain of tissue culture-adapted ATCC-Wa strain of human rotavirus and a novel inter-species variable domain in its C-terminus. AB - Complete nucleotide sequence of the tissue culture-adapted ATCC-Wa strain of human rotavirus NSP4 was determined. Sequence analysis detected two alternate forms of the gene with a nucleotide difference at position 331 (A or G) in the coding sequence (NSP4-A or NSP4-G) leading to a change from neutral glutamine97 in NSP4-A to a positively charged arginine97 in NSP4-G originating from the same ATCC-Wa preparation. In addition to this, both forms of ATCC-Wa NSP4 revealed three mutations at nucleotide positions 88 (T to C), 142 (C to T) and 572 (G to A), when compared to the previously reported NSP4 sequence from virulent Wa strain. The former two mutations were in the coding sequence and resulted in a leucine16 to serine16 and a proline34 to leucine34 change, while the third mutation was in the 3' non-coding region of the gene. The two amino acid changes may contribute to the 'tissue culture-adaptation' of ATCC-Wa strain. The ATCC-Wa NSP4 sequence was found to differ from the previously reported NSP4 sequence of attenuated Wa strain by lacking a mutation at 133 (T to C), though the mutations at 88 and 142 were present in both strains. Furthermore, comparison of deduced amino acid sequence of NSP4 from human, bovine, porcine and simian rotavirus strains identified a seven-residue (positions 135-141) inter-species variable domain in its C-terminal region. PMID- 11043942 TI - Apricot latent virus: a new species in the genus Foveavirus. AB - Extraction of viral double-stranded RNA from peach leaves infected with Apricot latent virus (ALV) followed by molecular cloning of synthesized cDNA and its sequencing, suggested that ALV is a new virus, whose coat protein (CP) coding region contains Apple stem pitting virus (ASPV)-related sequences. The sequenced portion of the ALV genome (1,444 nt) includes the putative CP gene and the 3' non translated region. The 5' portion of this fragment (1-651 nt) is highly distinct whereas the 3' portion is 77% identical to the corresponding region of ASPV. Molecular hybridization experiments using a cRNA probe to ASPV with ALV-infected leaf tissue extracts also revealed that the genome of ALV contains nucleotide sequences related to that of ASPV. Western blots of tissue extracts indicated that ALV coat protein reacted with polyclonal antiserum against ASPV; however, the ALV CP differs in size from that of ASPV. ALV was graft-transmitted to several Prunus rootstocks. Based on the available sequence data, serological observations and bioassays we propose that ALV is a new species in the genus Foveavirus, typified by ASPV. ALV-specific PCR-primers and viral-specific cRNA probes developed in this investigation may be useful for detecting the virus and for studying its epidemiology and geographical distribution. PMID- 11043943 TI - Presentation of antigenic sites from foot-and-mouth disease virus on the surface of baculovirus and in the membrane of infected cells. AB - We describe the construction of recombinant baculoviruses displaying on their surface and in the membrane of infected cells the small, immunodominant antigenic site (site A) or the large polyprotein (P1) coding for the four structural proteins of foot-and-mouth disease virus (FMDV). The coding sequences were inserted in the amino-terminus of gp64, the major glycoprotein of the baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV). Following infection of insect cells with the recombinant baculoviruses, the cellular localization of the chimaeric proteins as well as their presence in the surface of extracellular viruses was assessed by immunofluorescence microscopy and Western blot. The antigenicity of the recombinant viruses was studied by competitive ELISAs, which showed that although both recombinant viruses were able to compete with FMDV specific monoclonal antibodies (MAbs), their patterns of reactivity were different. The results suggest that this eukaryotic display system could be an alternative method of presentation of foreign antigens in a multimeric form as a new approach to biosynthetic vaccines. PMID- 11043944 TI - Genome replication of Newcastle disease virus: involvement of the rule-of-six. AB - We examined replication of Newcastle disease virus (NDV) by using minigenomes consisting of the 3' leader and 5' trailer regions of NDV flanking a reporter gene encoding secreted placental alkaline phosphatase (SEAP). Negative-sense minigenome RNA was generated from transfected plasmid DNA by means of in vivo transcription. Subsequent replication of minigenome RNA was determined either after infection with NDV helpervirus or after contransfection with helperplasmids that expressed the essential viral replication proteins NP, P, and L. In both systems, efficient replication of minigenome RNA was observed only if the genome size was a multiple of six nucleotides. Hence, in these systems, replication of NDV minigenome RNA's is strictly dependent on the rule-of-six. When the supernatant from helpervirus-infected, transfected cells was used to infect fresh monolayers, efficient transfer of SEAP activity by virus-like particles was observed only if the size of the minigenome RNA obeyed the rule-of-six. However, after several serial passages, we also observed efficient transfer of SEAP activity by virus-like particles derived from minigenome RNA's that did not obey the rule-of-six. Evidence was obtained which indicated that successful replication of these minigenomes was not due to a change in genome size. PMID- 11043945 TI - Human astrovirus isolation and propagation in multiple cell lines. AB - Laboratory adapted human astrovirus serotypes 1 through 7 were tested for growth in 15 human, 7 simian, and 10 other non-primate mammalian cell lines. Propagation of all seven serotypes was successful in the human cell lines Caco-2, T84, HT-29, and in the African green monkey kidney cell line MA-104. Both primary and secondary African green monkey kidney cells were more effective than Rhesus monkey kidney cells for cultivation of astrovirus. Except for human foreskin cells, all of the other human and simian cell lines supported growth of at least one astrovirus serotype. The only non-primate cell line that permitted sustained passage of astroviruses was the BHK-21 (C13) cell line for astrovirus serotype 2. Seventeen human stool specimens that had previously been shown to be astrovirus positive by ELISA were cultured in Caco-2, T84, HT-29, SK-CO-1, PLC/PRF/5, MA 104, and VERO cells. Caco-2 cells (13 isolates), T84 cells (12 isolates) and PLC/PRF/5 cells (12 isolates) were the cell lines most effective for isolation of human astroviruses from clinical stool specimens. By immunofluorescent staining of infected cells, culturing of the same 17 specimens in shell vials for 18 h was positive for astroviruses in all 17 specimens in Caco-2 cells, 12 in T84 cells, and 7 in PLC/PRF/5 cells. Shell vial assay is suitable as a rapid and sensitive culture technique for detection of astroviruses in clinical specimens. PMID- 11043946 TI - The nucleotide sequence of Indian peanut clump virus RNA 2: sequence comparisons among pecluviruses. AB - The RNA-2 molecule of an isolate of the L serotype of Indian peanut clump virus (IPCV) was shown to consist of 4,290 nucleotides with five open reading frames (ORF). The arrangement of the ORFs resembled that in RNA-2 of Peanut clump virus (PCV) from West Africa. The proteins encoded by the ORFs in IPCV-L RNA are between 32% and 93% identical to those encoded by PCV RNA. Partial sequence data for the RNA-2 of isolates of the H and T serotypes of IPCV show that the coat and P40 proteins encoded by the 5'-most ORFs of RNA-2 of IPCV-L, IPCV-H and IPCV-T are as similar to each other as any is to the corresponding proteins of PCV. A conserved motif 'F-E-x6-W' is present near the C-termini of the coat proteins of all three IPCV serotypes and of PCV, as it is in the coat proteins of other viruses that have rod-shaped particles, such as Tobacco mosaic virus and Tobacco rattle virus. The results support the distinction of IPCV and PCV as separate virus species, but also raise the question of how the serotypes of IPCV should be classified. PMID- 11043947 TI - Recombination between a 3-kilobase tobacco mosaic virus transgene and a homologous viral construct in the restoration of viral and nonviral genes. AB - Transgenic plants harboring various plant virus sequences have shown resistance to viral infections. An environmental risk associated with the use of these plants is the possibility of forming a novel virus by recombination between challenging viruses and transgenic viral mRNA. Two experiments were designed using tobacco mosaic virus (TMV) vectors and transgenic Nicotiana benthamiana to determine if recombinant viral RNA would be detectable. N. benthamiana was transformed with a nontranslatable portion of a TMV viral vector including part of the replicase gene, the movement protein gene, a gene for green fluorescent protein (GFP), and the coat protein gene. When transformed plants were inoculated with a TMV vector coat protein mutant which could not move efficiently through the host, recombinant RNA was detected in 32% of the infected plants, although virions were not detected. When transformed plants were infected with a TMV vector with a normal coat sequence but three base changes in the GFP sequence, no recombinant RNA or virions were detected. Thus, recombinant RNA between TMV RNA and host mRNA did not accumulate to detectable levels under nonselective conditions, and though recombinant RNA did accumulate in the presence of selective pressure, an encapsidated recombinant viral population did not develop. PMID- 11043948 TI - Detection of herpesviral sequences in tissues of green turtles with fibropapilloma by polymerase chain reaction. AB - An alpha-herpesvirus has been associated recently with green turtle fibropapilloma (FP). To further clarify the role of this newfound green turtle herpesvirus (GTHV) in the pathogenesis of FP, various normal-appearing tissues and organs (including skin, eye, brain, heart, liver, spleen, intestine, lung, kidney, nerve, gonad, tongue, gall bladder, urinary bladder, thyroid and peripheral blood mononuclear cells (PBMC) from blood) and tumor tissues from 19 green turtles (Chelonia mydas) with FP, and tissues from three green turtles without FP, collected during 1997 to 1999 in the Hawaiian Islands, were tested for GTHV sequences by nested polymerase chain reaction (PCR), using GTHV-specific oligonuclotide primers. GTHV sequences were detected in all tumors (51/51) and most tissues (133/167) of tumored turtles. By contrast, such sequences were undetectable in tissues (0/28) of three non-tumored turtles. Analysis of GTHV sequences detected in different tissues and tumors revealed a low degree of genetic diversity (<1%). The wide distribution of this newfound herpesvirus in tumors and tissues of tumored green turtles and its absence in tissues of non tumored turtles, argues for an etiologic role in FP. PMID- 11043949 TI - Indian citrus ringspot virus: a proposed new species with some affinities to potex-, carla-, fovea- and allexiviruses. AB - An isolate of Indian citrus ringspot virus from Kinnow mandarin in northern India had flexuous particles with evident cross-banding and a modal length of 650 nm. It was mechanically transmitted to five herbaceous hosts including Phaseolus vulgaris cv Saxa, in which it became systemic. In thin sections, virus particles were observed in the cytoplasm of parenchyma cells but no specific inclusions were seen. The virus was purified from infected Saxa bean leaves and an antiserum prepared. There was no serological cross-reaction with representative allexi-, capillo-, potex- and trichoviruses, except a faint one-way reaction with Potato virus X. Purified virus yielded a major band, the presumed coat protein (CP), of about 34 kDa, and a single ssRNA of about 7.5 kb, which was infectious. Two ORFs encoding putative proteins of 34 kDa and 23 kDa were located in the 3' part of the RNA. The product of the 34 kDa ORF was confirmed as the CP by expression in E. coli. The derived amino acid sequence of the CP contained some short motifs similar to those of potex-, fovea-, carla- and allexiviruses but otherwise there was no strong similarity to any of these. The 23 kDa ORF contained a zinc finger like sequence, as in similar ORFs in carla- and allexiviruses but overall amino acid homology with these was low. The virus does not appear to fall into any known genus. A new species is proposed. Serological and molecular diagnostic reagents were prepared. PMID- 11043950 TI - Hepatitis E virus infection in chimpanzees: a retrospective analysis. AB - Different patterns of disease were observed among 11 chimpanzees who were inoculated intravenously with hepatitis E virus (HEV) positive fecal specimens from four different outbreaks (Nepal 1981, Uzbekistan 1981, Pakistan 1985, and Mexico 1986). Five chimpanzees had marginal or no liver enzyme elevations within 70 days of inoculation. Two of the chimpanzees had limited viremia, but did not produce detectable antibody. The four remaining chimpanzees had liver enzyme elevations, viral shedding, viremia, seroconversion to anti-HEV, and detectable HEV antigen in liver biopsy specimens. These results may reflect the range of infection patterns that develop in humans after natural exposure to the HEV. PMID- 11043951 TI - Characterisation of a recently isolated lyssavirus in frugivorous zoo bats. AB - In July 1997 a lyssavirus was isolated in Denmark from a colony of Egyptian flying foxes (Rousettus aegyptiacus) originating from a Dutch zoo. Sequencing of a 400 nucleotides coding region of the nucleoprotein and of a major part of the G protein ectodomain encoding region of the newly isolated virus, revealed a very high similarity with European Bat Lyssavirus subtype 1a (EBL-1a). For characterisation of the recently isolated lyssavirus in frugivorous zoo bats, 16 frugivorous bats (Rousettus aegyptiacus) of the same colony and 80 mice were experimentally infected with the Rousettus isolate or with a well defined EBL-1a strain isolated from a Dutch insectivorous bat (Eptesicus serotinus). Inoculation viruses were titrated in mice to determine LD50's of both isolates. Clinical signs of inoculated bats were recorded during 6 weeks. After showing neurological signs or at the end of the experimental infection all animals were euthanized. During the experimental infection sera and various tissues of inoculated bats were collected. Immunoassays, mouse inoculation tests (MIT) and polymerase chain reaction (PCR) were employed for detection of lyssavirus specific antibodies, antigen or RNA. Five bats inoculated with the Rousettus isolate and 2 bats inoculated with the Eptesicus isolate showed neurological signs. The remaining 9 bats survived and cleared the virus; at least under the detection limit of the used assays. Despite a much higher pathogenicity of the Rousettus isolate observed in mice, LD25's in bats were quite the same for the 2 isolates. The pathogenicity of both isolates suggested that like many other mammals, Rousettus aegyptiacus bats could be victims of lyssavirus infection besides reservoir hosts of infectious EBL1a. There was no significant difference in detecting the different lyssavirus isolates in Rousettus aegyptiacus bats. An employed immunoperoxidase staining (IP) method was very useful for sensitive detection and localization of lyssavirus antigen in histologic preparates. PMID- 11043952 TI - Molecular characterization and phylogenetic analysis of the Harrisina brillians granulovirus granulin gene. AB - Well over 100 isolates of granulosis viruses (GVs), genus Granulovirus (family Baculoviridae), have been reported, all from lepidopterous insects. Three types of GVs are recognized, those of Type 1, which attack the fat body, Type 2, which attack most tissues, and Type 3, which attack only the midgut epithelium. To determine whether a correlation exists between tissue tropism and lepidopteran family phylogeny, the granulin gene of the Harrisina brillians (HbGV), a virus that attacks the midgut epithelium of H. brillians (family Zygaenidae) was cloned, sequenced, characterized, and compared with granulin genes of GVs that attack species of Tortricidae, Pieridae, and Noctuidae. The HbGV granulin gene encoded a peptide of 248 amino acids with a predicted Mr of 29.6 kDa, and shared a significant level of homology with other granulin (81-95% identical and 90-98% similar) and polyhedrin (49-58% identical and 62-72% similar) proteins. Phylogenetic analyses based on granulin and polyhedrin genes as well as on their 5'-untranslated sequences (5'-UTSs) indicated that HbGV was more closely related to GVs isolated from the tortricids, Cryptophlebia leucotreta (ClGV), Cydia pomonella (CpGV) and Choristoneura fumiferana (CfGV) than to other GVs and NPVs. This analysis provides preliminary evidence for a correlation between GV tissue tropism and the phylogeny of lepidopteran families, suggesting that GVs attacking species of Tortricidae and Zygaenidae are ancestral to those attacking species of the family Noctudiae. PMID- 11043953 TI - Mapping of functional domains on the influenza A virus RNA polymerase PB2 molecule using monoclonal antibodies. AB - Monoclonal antibodies against the PB2 of A/Puerto Rico/8/34 (A/PR/ 8/34) (H1N1) were prepared in order to define the functional domains of the RNA polymerase of influenza virus. The fifteen monoclonal antibodies that were generated were divided into 4 groups on the basis of ELISA binding to PB2 or its peptide fragments. Six Group I antibodies that bound to the PB2 N-terminal region (amino acids 1-104) did not inhibit transcription by the viral ribonucleoprotein complex. A single Group II antibody recognizing the region of amino acids 206-259 inhibited ApG-primed transcription. Groups III and IV antibodies bound to the C terminal region of amino acids 660-759. Of these, Group III antibodies inhibited transcription. The present results identify multiple monoclonal antibody binding domains in PB2, two of which, when bound by antibodies, negatively affect viral RNA transcription. PMID- 11043954 TI - Relative localization of viroplasmic and endoplasmic reticulum-resident rotavirus proteins in infected cells. AB - Replication of rotaviruses, whose capsid is constituted by three concentric layers of proteins, occurs in large cytoplasmic inclusions, termed viroplasms. Subviral, double-layered particles bud from viroplasms to the adjacent endoplasmic reticulum (ER), where the outermost protein layer, formed by VP4 and VP7, is assembled. To better understand the morphogenetic process of the virus, we analyzed the relative distribution of viroplasmic and ER-resident viral proteins. Using double immunostaining and confocal microscopy we observed an extensive co-localization between the ER proteins NSP4 and VP7, and the cytoplasmic protein VP4. These three proteins were found to be organized mostly as ring-like or semicircular structures in close association with viroplasms, except for VP4 which displayed in addition, a filamentous distribution. The observations reported in this study underscore the highly organized nature of rotavirus morphogenesis. PMID- 11043955 TI - Characterisation of two citrus apscaviroids isolated in Spain. AB - Sequence variability in the PCR amplified cDNAs from two citrus apscaviroid isolates CVd-Ia and CVd-IIId from Spain, was analysed. CVd-IIId sequence was shown to be identical to previously described CVd-III sequences and no important variability was encountered within the viroid population. Conversely, CVd-Ia displayed population heterogeneity as shown by SSCP analysis, Sal I restriction site polymorphism and sequences of 27 CVd-Ia cloned DNAs. The CVd-Ia genomic heterogeneity is characterised by two major subpopulations with the most divergent sequences, and by the presence of individual variants, making a sequence continuum between the two major groups. Most sequence variations are clustered in the left part of the viroid molecule. PMID- 11043956 TI - ICTV and the Virology Division News. PMID- 11043957 TI - AV: show or substance? PMID- 11043958 TI - Complications of common selective spinal injections: prevention and management. AB - Selective spinal injections are being performed with increasing frequency in the management of acute and chronic pain syndromes. Because these procedures require a needle to be placed in or around the spine, there is always a risk of complications. For this reason, prevention, early recognition, and management of complications are paramount to patient care. Physician training and patient preparation and monitoring are required to maximize the safety and efficacy of the specific spinal procedures. This focused review article discusses the primary general and specific complications of spinal injection procedures as well as treatments. PMID- 11043959 TI - Early versus delayed treatment of enchondroma. AB - Enchondromata are among the most common primary neoplasms of the hand, which often present as pathologic fractures. The purpose of this study is to determine whether there are any differences between cases in which both fracture and tumor were treated primarily and those in which tumor treatment was delayed. We reviewed a total of 16 cases; six were treated immediately, 10 were delayed. The immediate treatment group had four complications, the delayed group had one. The theoretical advantages of immediate treatment include a decrease in both the period of disability and delay of definitive diagnosis. This study supports that supposition. However, we did note a significantly higher complication rate for the immediate treatment group (67% versus 10%). Our results indicate that while there is an apparent decreased disability period, there may be reason for caution in immediate treatment of both the fracture and the tumor in pathologic fractures through enchondromata. PMID- 11043960 TI - Radiographic analysis to determine the treatment outcome in developmental dysplasia of the hip. AB - The purpose of this retrospective study was to determine whether pretreatment radiographic findings of patients less than 6 months old with developmental dysplasia of the hip can predict treatment outcome. Medical records and radiographs were reviewed in a cohort of 35 patients (44 hips) less than 6 months old with a diagnosis of hip dislocation at one institution. Using pretreatment anteroposterior radiographs, we measured the superior gap (distance between the proximal metaphysis to Hilgenreiner's line) and the medial gap (distance between the femoral calcar and the lateral pelvic wall at that level) and analyzed these data along with the type of splint used (Pavlik harness verses other abduction orthosis). Medical records and radiographs from an aged-matched cohort of 20 patients (23 hips) treated at a second institution were analyzed by using the same clinical and radiographic criteria of hip dislocation. Of the 44 dislocated hips, 29 failed with a trial of splintage (66%). Of the 23 dislocated hips from the second institution, 16 failed a trial of splintage (70%). Statistical analysis evaluating the age of the patient, medial and superior gaps, and harness type revealed that an abnormal superior and medial gap consistently predicted success or failure of splintage. We concluded that infants with pretreatment radiographs revealing a superior gap equal to or less than 3 mm, or a medial gap equal to or greater than 10 mm should not be treated with an initial trial of splintage because failure is likely. PMID- 11043961 TI - Anatomic study of the cervicothoracic spinal nerves and their relation to the pedicles. AB - Twelve cadavers were dissected for the study of the cervicothoracic junction. The results showed that the mean heights and widths of the ganglia tend to decrease from the C-6 to T-4 nerve. The mean distances between the dura and the ganglion and the mean spinal nerve angles increased consistently from C-5 to T-4. The mean distances from the spinal nerves to the superior and inferior pedicles ranged 0.8 2.3 mm. It was noted that the mean value was significantly greater for the distance from the spinal nerve to the superior pedicle than that to the inferior pedicle for the spinal nerves C5-7 (P< or =.05). This information, in conjunction with imaging studies, may minimize spinal nerve injury during posterior pedicle screw fixation in the cervicothoracic spine. PMID- 11043962 TI - Synovial chondromatosis of the shoulder associated with osteoarthritis: conservative treatment in two cases and review of the literature. AB - Periarticular synovial chondromatosis associated with osteoarthrosis is a rare condition that is more frequently seen in the lower extremity than in the upper extremity. In patients who have synovial chondromatosis of the joints of the lower extremity and are symptomatic, the traditional method of treatment has included open or arthroscopic synovectomy and removal of loose bodies. In cases involving the upper extremity, especially in the shoulder, patients have variable disability and may be treated successfully without surgery. Two patients who presented to our practice with shoulder symptoms due to synovial chondromatosis were treated successfully without surgery. In both patients nonoperative treatment consisting of activity modification, nonsteroidal anti-inflammatory medication, and cryotherapy as needed led to a good result without surgical intervention. PMID- 11043963 TI - Extensor tendon rupture secondary to the AO/ASIF titanium distal radius plate without associated plate failure: a case report. AB - Complications with open reduction and internal fixation of distal radius fractures have led to the development of the AO/titanium Pi-plate. This was designed to be a low-profile plate to decrease its intrusion into surrounding soft tissues. Recent case reports are revealing an association of tendon ruptures with failures of this plate. This case report demonstrates extensor tendon rupture without plate failure or prominent screw heads. PMID- 11043964 TI - Synovial chondromatosis of the shoulder. AB - We describe a case of synovial chondromatosis involving the shoulder. Presenting symptoms, radiographic features, intraoperative findings, and treatment options are discussed. PMID- 11043965 TI - Fibrolipoma of the median nerve: a case report and review of the literature. AB - A 38 year-old patient presented with right median nerve distribution paresthesias. Electrodiagnostic studies confirmed severe carpal tunnel syndrome. A palmar mass prompted a magnetic resonance imaging scan, which suggested a fibrolipoma of the median nerve. Carpal tunnel release resulted in resolution of preoperative pain and paresthesias. We review the literature dealing with this primary nerve tumor. PMID- 11043966 TI - Osteochondroma of the distal clavicle: an unusual cause of rotator cuff impingement. AB - Rotator cuff impingement symptoms may arise from intrinsic pathologic shoulder lesions related to instability or from extrinsic factors, primarily involving the coracoacromial arch. This report describes 2 unusual cases in which extrinsic impingement resulted from a distal clavicular osteochondroma. In both cases symptoms resolved after resection of the osteochondroma. PMID- 11043967 TI - Reduction of skiing-related anterior shoulder dislocation using Kocher's method without traction. AB - We evaluated the use of Kocher's original method (without humeral traction) for reduction of acute anterior glenohumeral dislocation in 28 alpine skiers and snowboarders at a single ski area during the 1995-1996 ski season. In all cases, reduction was begun within 1 hour of the acute injury. The Kocher method alone was successful in 23 (82%) patients. Of the patients having a successful reduction by means of the original Kocher technique, the mean reduction time was less than 5 minutes, and 9 (39%) of the reductions were achieved in less than 1 minute. Only 1 patient experienced discomfort significant enough to require analgesia, and no patients required sedation. The complication rate was minimal, with 1 patient developing hyperesthesia in the axillary nerve distribution; no fractures of the humerus or glenoid resulted from the reduction technique. PMID- 11043968 TI - Diffuse idiopathic skeletal hyperostosis with ossification of the posterior longitudinal ligament. PMID- 11043969 TI - Pathomorphological variations of the AIDS-associated progressive multifocal leukoencephalopathy. AB - Pathological analysis of 20 cases of the progressive multifocal leukoencephalopathy (PML) appearing in the course of acquired immune deficiency syndrome (AIDS) is presented. PML occurred in 10% of all AIDS cases, collected in the period from 1987 to 1999. PML appeared either as the only brain pathology or accompanied HIV-related brain alterations isolated or concomitant with one or several opportunistic infections and/or neoplastic growth (malignant lymphoma). Basing on the pathomorphological picture and clinical symptomatology early, atypical and severe forms of the disease were distinguished. All of them were characterized by typical PML demyelination with oligodendroglial and astrocytic pathology. The group with early changes revealed widespread, multifocal myelin alterations of a moderate intensity with predominant oligodendroglial abnormalities and less advanced astrocytic changes. Atypical form of the disease was represented by cases with unifocal changes, although containing all key elements of PML pathology. The leading pathological feature of the severe form of the disease consisted in a particular intensity of the demyelination, resulting in tissue destruction often with its cavitation, with typical glial reaction and intense macrophage and lymphocytic infiltration. The other distinguishing feature consisted in strong topographic prevalence of the pathological process either to brain hemispheres or cerebellum. Differences of PML pathology in the course of AIDS as compared with non-AIDS cases are discussed. Due to the relatively high frequency of cases of isolated or strongly predominant involvement of cerebellum, separation of the cerebellar form of the disease has been suggested. PMID- 11043970 TI - Myositis specific antibodies: frequency in different populations. AB - Myositis specific autoantibodies (MSA) are the most specific markers of idiopathic inflammatory myopathies (IIM). There is no evidence of presence MSA in patients with other neuromuscular or connective tissue diseases. We compared the frequency of MSA in two groups of IIM patients, one from Poland and one from North America and found no significant statistical differences (21% and 25% respectively). There was a significant difference between the occurrence of immunological marker PM-Sci in scleromyositis patients (22.85% in group I and 7.1% in group II). This figure was also greater than those previously reported in North Americans (2-10%) and Japanese (extremely seldom). These findings confirm the association between MSA and several homogenous clinical syndromes: antisynthetases with the antisynthetase syndrome, anti-SRP with severe, resistant to treatment myositis, anti-Mi-2 with classic, benign dermatomyositis. They underscore the importance of including MSA in the routine diagnostic workup of IIM. PMID- 11043971 TI - Morphometric analysis of axons in the minute multiple sclerosis lesions and shadow plaques in patients with multiple sclerosis. AB - The objective of the present study was to quantitatively detect axons in the minute multiple sclerosis (MS) lesions and in shadow plaques, taking into consideration the relapsing-remitting(R-R) and secondary progressive(SP) stages of MS. The brain tissue of 12 patients deceased due to MS was investigated. An image-computerized analysis was made for measurements of axons. Based on the findings we concluded that damage to axons appears in both the minute MS lesions and in shadow plaques. Demyelination and ineffective (too late or too slow) remyelination seemed to be very important factors in axonal damage. Irreversible damage to axons may appear in both the secondary progressive and relapsing remitting stages of MS, causing permanent neurological deficits, irrespective of the duration of the disease. PMID- 11043972 TI - Major histocompatibility complex class II (MHC II) expression during the development of human fetal cerebral occipital lobe, cerebellum, and hematopoietic organs. AB - In adults, under physiological conditions proteins of the major histocompatibility complex, class II (MHC II) molecules are synthesized and then presented on the surface of the cells known under a common name as antigen presenting cells (APCs). Dendritic cells (DCs), microglia, macrophages, ameboid microglia and lymphocytes B are qualified as APCs. The aim of present study was to evaluate the expression of MHC II molecules in the central nervous system (CNS) and hematopoietic organs during the fetal development. Observations were made on the cerebral occipital lobe, cerebellum, thymus, spleen and liver of 30 normal human fetuses, between 11 and 22 week of gestation (GW). Histological, histochemical and immunohistochemical techniques were used to identify cells with expression of MHC II molecules. In the brain, MHC II molecules were detected on macrophages/ameboid microglia in meninges, choroid plexus and single cells of ramified microglia in deeper layers of the cortex and white matter. In the other organs besides macrophages and dendritic cells, MHC II molecules were also immunopositive in thymic epithelial cells, and in the spleen and liver also in other cells of stroma and lobule. The expression of MHC II molecules on so extensive population of cells, at an early stage of the fetal development, may evidence their significant involvement in histogenesis and morphogenesis. It seems that in adults the complex of MHC II with protein is originated from the foreign antigen. On the contrary, during normal fetal development the complex of MHC II with protein origins most probably from the fetus own structures. PMID- 11043973 TI - Intrasellar neuronal hamartoma associated with pituitary adenoma. Case report. AB - 46-year-old acromegalic women presenting high level of growth hormone (32 ng/ml) in the serum underwent surgery. The intrasellar tumor, 16 mm in diameter, has been removed. The biopsy material consisted of two types of closely adjacent and intermingled tissues, one of which was growth hormone positive acidophilic adenoma, the second component were haphazardly oriented ganglion cells of various size and shape, also multinuclear, with bundles of unmyelinated fibers. The cytoplasm and processes of ganglion cells were immunopositive for neurofilaments and for synaptophysin on cellular membranes and processes. There were none glial fibrillary acidic protein positive cellular elements. The authors discuss commonly used name of choristoma for this type of tumor and the problem of possible neurosecretory stimulation of pituitary adenoma by neuronal hamartoma. PMID- 11043974 TI - Xanthomatous changes in atypical and anaplastic meningiomas. Light and electron microscopic investigations. AB - Xanthomatous changes may occur in meningiomas of different histological type, however their incidence in combination with histological features of atypical or anaplastic meningioma has not been previously documented. In this report we present clinicopathologic, immunohistochemical and ultrastructural studies in the surgical cases of two atypical and three anaplastic meningiomas exhibiting prominent xanthomatous changes. In all tumors the xanthomatous cells were seen in association with typical meningioma structures such as meningothelial whorls or psammoma bodies as well as within the tumor parts displaying pleomorphism, patternless growth, increased cellularity, presence of necroses and mitoses or brain invasion. Ultrastructural study revealed a wide-range of lipid-containing cells, reflecting a continuum of gradual transition between polymorphic meningioma cells and xanthomatous cells. Commonly, the lipidized cells exhibited different degrees of plasmalemmal interdigitations and desmosomal junctions. Our study allowed us to confirm the meningothelial origin of xanthomatous cells in atypical and anaplastic meningiomas. Moreover, the ultrastructural observations of lysosomes in the majority of xanthomatous cells and the immunoreactivity for the CD68 antigen indicated their macrophage characteristics. It seems that a mixed meningeal/macrophage nature of xanthomatous cells can be related to the functional and structural multipotentiality of the primary leptomeningeal cells. PMID- 11043975 TI - Disseminated spinal and cerebral ependymoma with unusual histological pattern: clinicopathological study of a case with retrograde tumor spread. AB - The subject of this study is a case of anaplastic ependymoma originally arising from the central canal of the lower spinal cord followed by the 13 years history of events of upper spinal dissemination and retrograde intracranial spread. The specimens from four subsequent surgeries generally displayed the same microscopic features of neoplastic tissue and were consistent with the diagnosis of anaplastic ependymoma. The histological diagnosis was based upon the high cellularity, considerable nuclear atypia and pleomorphism, brisk mitotic activity, focally exhibited vascular endothelial proliferation and extensive necrosis. Apart from the typical pattern of ependymoma, the tumors contained areas composed almost entirely of large, uniform clear cells or pseudogemistocytes indicating the morphological heterogeneity of neoplastic cells population. The surgical specimens from four surgical resections shared light microscopic similarities suggesting spinal and intracranial dissemination from the primary spinal tumor. Since the retrograde spread via the cerebrospinal fluid (CSF) pathway is extremely rare, the authors of this study discuss the mechanism of such way of tumor metastases. PMID- 11043976 TI - Zentralblatt fur Bakteriologie--100 years ago: the isolation of Shigella flexneri. PMID- 11043977 TI - Apoptosis as a common bacterial virulence strategy. AB - The comparison of common strategies used by bacterial pathogens to overcome host defenses provides us with the opportunity to analyze the biology of pathogenicity, as well as point out the unique interactions between a particular pathogen and its host. Here we compare and contrast apoptosis induced by three enteric pathogens, Salmonella, Shigella, and Yersinia. We point out that all three enteric pathogens induce apoptosis in macrophages in vitro, but the proposed mechanisms are quite different. Yersinia induces apoptosis by inhibiting the translocation of the transcriptional activator, NF-kappaB, into the nucleus, which results in the suppression of TNFalpha production; whereas Salmonella- and Shigella-induced apoptosis depend on the activation of caspase-1 (casp-1). The result of casp-1 activation is to induce apoptosis as well as to process the proinflammatory cytokines, pro-IL-1beta and pro-IL18 into their mature bioactive forms. Thus, in contrast to Yersinia, Salmonella and Shigella-induced apoptosis results in a proinflammatory cascade. PMID- 11043978 TI - Host-pathogen interactions in mycoplasma pathogenesis: virulence and survival strategies of minimalist prokaryotes. AB - Despite their very small genomes mycoplasmas are successful pathogens of man and a wide range of animal hosts. Because of the lack of effective therapeutics and vaccines, mycoplasma diseases continue to be a significant problem for public health as well as livestock production with major socio-economic consequences worldwide. Recent outbreaks and epidemiological studies predict that the incidence of human and animal mycoplasma diseases might increase which indicates the urgent need to develop new approaches for prevention and therapy. Development of such reagents, however, requires a solid understanding of the molecular biology of mycoplasma infections. Knowledge in this field has considerably increased during the past decade since new techniques have been developed and adapted to mycoplasmas that allow these organisms to be studied at the molecular level. Research on the two human pathogens Mycoplasma pneumoniae and Mycoplasma genitalium of which the genome sequences have recently been completed as well as the substantial number of studies carried out on the AIDS-associated mycoplasmas, Mycoplasma penetrans and Mycoplasma fermentans, has led the way, but a number of animal mycoplasmas are becoming increasingly appreciated as models for the study of the molecular basis of mycoplasma diseases. This review summarizes and highlights some of the recent findings concerning the molecular interactions that occur between pathogenic mycoplasmas and their hosts, both the common strategies as well as some unique approaches evolved by particular mycoplasma pathogens, including adherence to and uptake into non-phagocytic host cells, as well as mechanisms of escaping the host immune system. PMID- 11043979 TI - Bacterial adhesins: function and structure. AB - Specific adhesion to host tissue cells is an essential virulence factor of most bacterial pathogens. The fundamental processes that determine bacterial attachment to host tissue surfaces are mediated by microbial adhesins. Host specificity and tissue tropism are characteristics exhibited by different bacteria and are determined (at least in part) by the interaction between adhesins and their complementary receptors on host cell surfaces. A detailed picture of how bacteria are able to target to various receptors is emerging. A large number of bacterial adhesins with individual receptor specificities have been identified. Furthermore, recent research has shown that individual adhesins are prone to rapid microevolution that results in changes in the receptor specificity of individual adhesins. Microbial adhesins are often assembled into complex polymeric organelle structures, however non-organelle adhesins linked to the cell surface as monomers or simple oligomers also exist. This review gives an overview of bacterial adhesins and focuses on some general aspects of their biogenesis and role in bacterial colonization of host cell surfaces and as virulence factors. PMID- 11043980 TI - Cloning and functional characterization of a 30 kb gene locus required for lipopolysaccharide biosynthesis in Legionella pneumophila. AB - The spontaneous Legionella pneumophila lipopolysaccharide (LPS) mutant 137, which did not bind the LPS-specific mAb 2625, was complemented with a genomic library from the parental wild-type strain. Transformants were screened for reconstitution of the wild-type LPS phenotype, able to bind mAb 2625. By this strategy, a 32,661 bp region comprising 30 open reading frames (Orfs) was identified. Orfs with significant homologies to genes encoding enzymes required for LPS or capsule biosynthesis of Gram-negative bacteria were located on the gene locus. The mutation of strain 137 could be assigned to a deletion of a cytosine residue in Orf 8. The protein encoded by Orf 8 exhibited homology to bacterial methyl-transferases. The L. pneumophila LPS gene locus included genes with deduced products likely to be involved in LPS core oligosaccharide biosynthesis (rmlA-D, rhamnosyl-transferases, acetyl-transferase) as well as LPS O-chain biosynthesis and translocation (mnaA, neuB, neuA, wecA, wzt, wzm). The neuA (Orf 25) and neuB (Orf 24) gene products were functionally characterized by complementation of the capsule negative E. coli K1 mutants EV5 and EV24, respectively. By introduction of the L. pneumophila neuA gene into E. coli EV5 and the neuB gene into EV24, expression of the K1 polysialic acid capsule could be restored. We, therefore, conclude that the biosynthesis pathway of legionaminic acid, the structural unit of the L. pneumophila Sg1 O-antigen, might be similar to the biosynthesis of sialic acid. Southern blot analysis indicated the entire gene locus to be present in L. pneumophila serogroup (Sg)1 strains, whereas only parts of the DNA stretch hybridized to DNA from Sg2 to Sg14 strains. PMID- 11043981 TI - Molecular characterization of a novel siderophore-independent iron transport system in Yersinia. AB - Enteropathogenic Yersinia enterocolitica can be divided into mouse lethal (biogroup 1 B serotypes O:8, O:13, O:20 and O:21) and mouse non-lethal (biogroups 2-4 serotypes O:3, O:5,27, O:9) strains. Mouse-lethality is associated with the presence of the high-pathogenicity island encoding the TonB-dependent ferric yersiniabactin uptake system. The present study reports on a TonB-independent and non-siderophore yersiniae ferric uptake system, yfu. Genetic and functional characterization of the yfu determinant revealed high relationship to the periplasmic-binding-protein-dependent Serratia marcescens ferric uptake system sfu. The yfu locus is common to all Yersinia species pathogenic for humans. Gene targeting of the yfu locus has demonstrated its importance for ferric iron acquisition in vitro. However, yfu is not required for mouse virulence of Y. enterocolitica serotype O:8. PMID- 11043982 TI - Yersinia enterocolitica 16S rRNA gene types belong to the same genospecies but form three homology groups. AB - The species Yersinia (Y.) enterocolitica consists of biochemically and serologically heterogeneous strains. A vernacular nomenclature divides these strains in 'European' and 'American' bioserotypes. We investigated six strains of each group by DNA-DNA hybridization, determination of G + C mol% content and sequence alignment studies. Based on different DNA-DNA hybridization values and the 16S rRNA gene sequences a division into two Yersinia enterocolitica subspecies is justified. We propose the names Yersinia enterocolitica subsp. enterocolitica for strains belonging to the 16S rRNA gene type represented by the Type strain ATCC 9610 and Yersinia enterocolitica subsp. palearctica for strains belonging to the 16S rRNA gene type of strain Y11 (DSMZ13030). PMID- 11043983 TI - Internalization of extraintestinal Escherichia coli O18 strains by epithelial cells is modulated by EGF, insulin, and effectors of transmembrane signal transduction. AB - Adhesion to and internalization into host cells is an essential step in the pathogenesis of various bacterial infections. Here we investigated the effects of growth factors on the internalization of Escherichia coli O18 strains isolated from patients with urinary tract infection (UTI) by human epithelial cells. A dramatic increase in the uptake of Escherichia coli was observed after treatment of epithelial cells with epidermal growth factor (EGF) and to a lower extent with insulin. EGF-dependent internalization can be suppressed by tyrosine kinase inhibitors suggesting an involvement of the receptor tyrosine kinases in the regulation of the endocytotic process. Inhibitors of phospholipase A2, lipoxygenase, and cyclooxygenase significantly decreased internalization of bacteria induced by EGF. Finally, the specific inhibitor of PI 3-kinases Wortmannin was shown to suppress completely the EGF-independent internalization. The data of this analysis indicate the involvement of several signaling paths in bacterial internalization of uropathogenic Escherichia coli O18 strains and contribute to the comprehension of the pathogenesis of recurrent UTI. PMID- 11043984 TI - Influence of pathogenicity islands and the minor leuX-encoded tRNA5Leu on the proteome pattern of the uropathogenic Escherichia coli strain 536. AB - The uropathogenic Escherichia coli strain 536 (O6:K15:H31) carries four distinct DNA regions in its chromosome, termed pathogenicity islands (PAIs I536 to IV536). Each of these PAIs encodes at least one virulence factor. All four PAIs are associated with tRNA genes. PAI I536 and PAI II536 can be spontaneously deleted from the chromosome by homologous recombination between flanking direct repeats. The deletion of PAI II536 results in the truncation of the associated gene leuX encoding the tRNALeu. This tRNA influences the expression of various virulence traits. In order to get a deeper insight into the role of PAI I536/II536 and of the tRNA5LeU for the protein expression, the protein expression patterns of Escherichia coli 536 and different derivatives were studied. Differences in the protein expression patterns of the wild-type strain Escherichia coli 536, its mutants 536-21 (PAI I536-, PAI II536-, leuX-), 536delta102 (PAI I536+, PAI II536+, leuX-) as well as of the strain 536R3 (PAI I536-, PAI II536-, leuX+) were analyzed by two-dimensional polyacrylamide gel electrophoresis and MALDI-TOF mass spectrometry. We identified about 39 different intracellular proteins whose expression is markedly altered in the different strain backgrounds. These differences can be linked either to the presence or absence of the PAI I536 and PAI II536 or to that of the tRNA gene leuX. The identities of 34 proteins have been determined by MALDI-TOF-MS. The identification of five proteins was not possible. The results suggest that proteome analysis is an efficient approach to study differences in global gene expression. The comparison of protein expression patterns of the uropathogenic E. coli strain 536 and different derivatives revealed that in this strain the expression of various proteins including those encoded by many housekeeping genes is affected by the presence of PAI I536 and Pai II536 or by that of the tRNA5Leu. PMID- 11043985 TI - Molecular cloning and targeted deletion of PEP2 which encodes a novel aspartic proteinase from Aspergillus fumigatus. AB - An aspartic proteinase PEP2 [EC 3.4.23.25] was purified from a cell wall fraction of Aspergillus fumigatus. The enzyme, which showed a broad range of activity from pH 2.0 to 7.0 and migrated as a single band of 39 kDa in SDS-PAGE, was not detected in the culture supernatant. A specific gene probe was designed on the basis of the N-terminal sequence of the native protein, and the PEP2 genomic and cDNA were isolated from corresponding libraries. The deduced amino acid sequence of PEP2 consists of 398 amino acids. A signal sequence of 18 amino acids and a proregion of another 52 amino acids were identified. The mature protein consists of 328 amino acids which include the two DTG-motifs of the active site common to almost all pepsin-like enzymes. PEP2 showed a 64% identity with the vacuolar proteinase A (PrA), of Saccharomyces cerevisiae, and an 88% identity with PEPE, an aspartic proteinase of Aspergillus niger. Recombinant PEP2 was overexpressed in Pichia pastoris and the active enzyme was secreted into the culture supernatant. Targeted deletion of PEP2 did not affect vegetative growth or cell and colony morphology. Identification of proteinases, such as PEP2, which are apparently associated with the Aspergillus cell wall raises new interest in these molecules with respect to their possible function in the pathogenesis of invasive aspergillosis. PMID- 11043986 TI - Assessment of genetic relatedness of vaginal isolates of Candida albicans from different geographical origins. AB - PCR fingerprinting with single non-specific primers was used to type vaginal isolates of C. albicans from Portugal, Angola, Madagascar, and two regions of Germany (Berlin and Munich). In addition to analysing isolates that exhibited the normal biotype of C. albicans, the study included atypical strains that failed to assimilate glucosamine and N-acetylglucosamine, which were isolated from women in Angola and Madagascar. A total of 212 strains of C. albicans were studied, representing 87 different multi-locus genotypes. The genotypes of strains from each geographical population were highly similar but not identical. There was one exception: a strain from Portugal grouped with the typical strains from Angola. The typical and especially the atypical populations from Africa displayed less genotype variation than the populations from Europe. The Portuguese samples exhibited the greatest genotypic heterogeneity. Distance analysis (UPGMA) revealed a statistically weak correlation between genotype and geographical origin of the C. albicans isolates. PMID- 11043987 TI - Infection of organotypic slice cultures from rat central nervous tissue with Trypanosoma brucei brucei. AB - We recently described a new procedure to grow nervous tissue as organotypic culture. The main feature of these slice cultures is to maintain a well preserved, three-dimensional organisation of the central nervous tissue. As these cultures can be kept for several weeks (up to three months), we have used this in vitro approach to study the complex interactions between host tissue and parasites during late stages of cerebral African trypanosomiasis. Light and electron microscopical studies, as well as electrophysiological recordings demonstrate that the structure and function of the nervous tissue is not severely affected even after several weeks of trypanosome infection. The presence of a large number of parasites does not seem to be deleterious to neuronal survival. Secondly, most of the trypanosomes are located around the periphery of the nervous tissue, but many of them also penetrate into the nervous parenchyma. Thirdly, trypanosomes with well-conserved morphology are found within the cytoplasm of glial cells, which in some cases were identified as astrocytes. These "intracellular parasites" seem to actively invade the target cells. Our study demonstrates that the presence of proliferating trypanosomes does not per se interfere with the neural activity of CNS tissues. Secondly, it provides, to the best of our knowledge, the first in vitro demonstration of intracellular forms of African trypanosomes. PMID- 11043988 TI - Chromosomal rearrangements affecting biofilm production and antibiotic resistance in a Staphylococcus epidermidis strain causing shunt-associated ventriculitis. AB - During two clinical courses of shunt-associated meningitis in a 3-month-old child, five multiresistant S. epidermidis isolates were obtained and analyzed with regard to biofilm production and antibiotic susceptibility. Three S. epidermidis strains, which were initially isolated from the cerebrospinal fluid, produced biofilms on polystyrene tissue culture plates. Following antibiotic treatment and subsequent exchange of the shunt system, sterilization of the CSF was achieved. However, after three weeks a relapse of the infection occurred. The two S. epidermidis isolates obtained now were biofilm negative, but showed an identical resistance pattern as those from the previous infection, except that resistance to rifampicin and increased mininal inhibitory concentrations of aminoglycoside antibiotics had emerged. DNA fingerprinting by PFGE indicated the clonal origin of all isolates. However, some DNA rearrangements and differences in the IS256-specific hybridization patterns could be identified in the isolates from the second infection period that led to altered biofilm formation and increased expression of aminoglycoside resistance traits. The data evidence that variation of biofilm expression occurs in vivo during an infection and highlight the extraordinary genome flexibility of pathogenic S. epidermidis. PMID- 11043990 TI - The effect of hyperglycemia on cocaine neurotoxicity and death in mice. AB - OBJECTIVE: Cocaine toxicity frequently manifests as seizures and status epilepticus. Frequently, dextrose is administered to patients with cocaine induced seizures. The objective of this study was to examine the effect of pre existing hyperglycemia on cocaine neurotoxicity and death in mice. METHODS: Swiss albino mice received intraperitoneal dextrose at a dose of 1 g/kg (12.5%) (hyperglycemic group, n = 98). The euglycemic group (n = 98) received an equal volume of 0.9% saline. After 60 minutes, all the animals received intraperitoneal cocaine at a dose of 90 mg/kg. The times to onset of ataxia, seizure, and death were recorded in seconds. Times to events were compared using a Kaplan-Meier method and results were compared using the logrank test. The overall percentage outcomes were compared using chi-square. RESULTS: The ataxia rates (hyperglycemic 97%, euglycemic 97%, chi(2) = 0, p = 1), seizure rates (hyperglycemic 85%, euglycemic 82%, chi(2) = 0.292, p = 0.589), and survival rates (hyperglycemic 62%, euglycemic 51%, chi(2) = 0.2514, p = 0.113) were similar between the groups. The survival following a seizure was significantly higher in the hyperglycemic group (hyperglycemic 57%, euglycemic 41%, chi(2) = 4.439, p = 0.035). The median ataxia time was earlier in the hyperglycemic group (190 sec) than in the euglycemic group (166 sec) (p = 0.031). Seizures occurred no earlier in the hyperglycemic group (331 sec) than in the euglycemic group (342 sec) (p = 0.207). Survival times were not different for the hyperglycemic group (9,133 sec) and the euglycemic group (7,593 sec) (p = 0.394). Survival times following seizures were not different for the hyperglycemic group (8,095 sec) and the euglycemic group (5,816 sec) (p = 0.0752). CONCLUSIONS: In mice with pre-existing hyperglycemia, ataxia occurred earlier and survival following cocaine-induced seizures was longer than for euglycemic mice. No significant difference in the overall percentage of seizures and death was detected. Pre-existing hyperglycemia had minimal effect on worsening cocaine toxicity in mice. PMID- 11043989 TI - Hypertonic saline treatment of severe hyperkalemia in nonnephrectomized dogs. AB - OBJECTIVES: To determine whether a hypertonic saline bolus improves cardiac conduction or plasma potassium levels more than normal saline infusion within 15 minutes of treatment for severe hyperkalemia. Previously with this model, 8.4% sodium chloride (NaCl) and 8.4% sodium bicarbonate (NaHCO(3)) lowered plasma potassium equally effectively. METHODS: This was a crossover study using ten conditioned dogs (14-20 kg) that received, in random order, each of three intravenous (IV) treatments in separate experiments at least one week apart: 1) 2 mmol/kg of 8.4% NaCl over 5 minutes (bolus); 2) 2 mmol/kg of 0.9% NaCl over one hour (infusion); or 3) no treatment (control). Using isoflurane anesthesia and ventilation (pCO(2) = 35-40 torr), 2 mmol/kg/hr of IV potassium chloride (KCl) was infused until conduction delays (both absent p-waves and >/=20% decrease in ventricular rate in 0.70) in the various domains. The internal consistency of the ER scale was also high (r = 0.85). CONCLUSIONS: The ER scale appears to be valid and reliable. It performs well when compared with previously psychometrically tested tools. It is a sensible, well-adapted tool for the teaching environment offered by EM. PMID- 11043998 TI - Does interview date affect match list position in the emergency medicine national residency matching program match? AB - OBJECTIVE: Some residency applicants believe that the date on which they interview with a residency program influences how the program ranks them in the National Residency Matching Program (NRMP). Therefore, the authors studied whether interview date affects match list position in the emergency medicine (EM) residency match. METHODS: Forty-four Accreditation Council for Graduate Medical Education (ACGME)-accredited EM residency programs participated in this multicenter study. The interview date and match list position were collected for each interviewee for the 1997-98 season. Programs were also asked about factors that might potentially bias interview date assignment. Statistical analyses were performed both with and without these programs included. Interview dates and match list positions were standardized into percentile date and percentile rank for each program and were compared using linear regression analysis. Scatterplots graphed interview date vs match list position. Two-sample t-tests compared interview dates for ranked and nonranked interviewees. RESULTS: Data were collected for 3,800 individual interviews; 14% of these resulted in unranked applicants. Twenty-three programs, representing 1,997 interviews, reported potential bias in their interview date assignment. Regression analysis revealed an R(2) of 0.018268 (correlation coefficient = 0. 1352, 95% CI = 0.0992 to 0.1617) for all programs, R(2) of 0.010626 (correlation coefficient = 0.1031, 95% CI = 0.0571 to 0.1485) for programs without reported potential bias, and R(2) of 0.02444 (correlation coefficient = 0.1563, 95% CI = 0.10887 to 0.20309) for programs with reported bias. Scatterplots revealed no linear correlation. Two sample t-tests for all programs, and programs with and without reported bias showed no significant difference in average interview date for ranked and unranked interviewees (both with p > 0.2). CONCLUSION: In this study, interview date for EM residency positions in the 1997-98 season did not affect match list position among ranked applicants. Moreover, interview date had nno effect on the decision to leave candidates unranked. PMID- 11043999 TI - The value of diversity in academic emergency medicine. PMID- 11044000 TI - Women in Academic Emergency Medicine/Diversity Interest Group position statement. PMID- 11044001 TI - Preventive care in the emergency department, Part I: Clinical preventive services -are they relevant to emergency medicine? Society for Academic Emergency Medicine Public Health and Education Task Force Preventive Services Work Group. AB - In 1998 the Society for Academic Emergency Medicine's (SAEM's) Board of Directors asked the SAEM Public Health and Education Task Force to develop recommendations for prevention, screening, and counseling activities to be conducted in emergency departments (EDs). The Task Force's work was divided into two phases: 1) a discussion of the rationale for preventive services in the ED, along with generation of a preliminary list of prevention activities that could be studied for ED implementation; and 2) a formal evidence-based review of topics chosen from the preliminary list, along with recommendations for ED implementation and further study. This paper represents Phase I of the project. Phase II, the formal evidence-based review and recommendations, is published separately in this issue. PMID- 11044002 TI - Preventive care in the emergency department, Part II: Clinical preventive services--an emergency medicine evidence-based review. Society for Academic Emergency Medicine Public Health and Education Task Force Preventive Services Work Group. AB - INTRODUCTION: Emergency departments (EDs) provide an opportunity to initiate preventive services for millions of Americans who have no other source for these services. OBJECTIVES: To identify primary and secondary preventive interventions appropriate for inclusion in routine emergency care and, secondarily, to recommend areas in which research into the efficacy and cost-effectiveness of interventions is needed. METHODS: Systematic reviews were performed on 17 candidate preventive interventions with potential applicability in the ED. All but one was selected from those reviewed by the U.S. Preventive Services Task Force (USPSTF). Each two-person review team followed a template that provided a uniform approach to search strategy, selection criteria, methodology appraisal, and analysis of the results of primary studies bearing on ED cost-effectiveness. Assigned proctors provided methodological guidance to the review teams throughout the review process. A grading scheme was developed that took into account the evidence and recommendations of the USPSTF supporting primary efficacy of the intervention and the level of evidence supporting ED application identified by the Society for Academic Emergency Medicine Public Health and Education Task Force (PHTF) review teams. RESULTS: Seventeen reviews were completed. The following interventions received an alpha rating, indicating that evidence is sufficient to support offering these services in the ED setting, assuming sufficient resources are available: alcohol screening and intervention, HIV screening and referral (in high-risk, high-prevalence populations), hypertension screening and referral, adult pneumococcal immunizations (age >/=65 years), referral of children without primary care physicians to a continuing source of care, and smoking cessation counseling. Interventions receiving a beta or gamma rating, indicating that existing research is not sufficient to recommend for or against instituting them routinely in the ED, include: identification and counseling of geriatric patients at risk of falls, Pap tests in women having a pelvic exam in the ED, counseling for smoke detector use, routine social service screening, depression screening, domestic violence screening, safe firearm storage counseling, motorcycle helmet use counseling, and youth violence counseling programs in the ED. Interventions not recommended for ED implementation (omega rating) include Pap test screening for women not having a routine pelvic exam, diabetes screening, and pediatric immunizations. CONCLUSIONS: A set of recommendations for prevention, screening, and counseling activities in the ED based on systematic reviews of selected interventions is presented. The applicability of these primary and secondary preventive services will vary with the different clinical environments and resources available in EDs. The PHTF recommendations should not be used as the basis of curtailing currently available services. This review makes clear the need for further research in this important area. PMID- 11044003 TI - SAEM diversity position statement. The SAEM Diversity Interest Group. PMID- 11044004 TI - The emergency department presentation of patients with active pulmonary tuberculosis. AB - OBJECTIVE: To determine the clinical presentation of emergency department (ED) patients with active pulmonary tuberculosis (TB). METHODS: This was a retrospective medical record review of adult patients, identified through infection control records, diagnosed as having active pulmonary TB by sputum culture over a 30-month period at an urban teaching hospital. The ED visits by these patients from one year before to one year after the initial positive sputum culture were categorized as contagious or noncontagious, using defined clinical and radiographic criteria. The medical records of patients with contagious visits to the ED were reviewed to determine chief complaint, presence of TB risk factors and symptoms, and physical examination and chest radiograph findings. RESULTS: During the study period, 44 patients with active pulmonary TB made 66 contagious ED visits. Multiple contagious ED visits were made by 12 patients (27%; 95% CI = 15% to 43%). Chief complaints were pulmonary 33% (95% CI = 22% to 46%), medical but nonpulmonary 41% (95% CI = 29% to 54%), infectious but nonpulmonary 14% (95% CI = 6% to 24%), and traumatic/orthopedic 12% (95% CI = 5% to 22%). At least one TB risk factor was identified in 57 (86%; 95% CI% = 76 to 94%) patient visits and at least one TB symptom in 51 (77%; 95% CI = 65% to 87%) patient visits. Cough was present during only 64% (95% CI = 51% to 75%) of the patient visits and hemoptysis during 8% (95% CI = 3% to 17%). Risk factors and symptoms that, if present, were likely to be detected at triage were foreign birth, homelessness, HIV positivity, hemoptysis, and chest pain. CONCLUSIONS: Patients with active pulmonary TB may have multiple ED visits, and often have nonpulmonary complaints. Tuberculosis risk factors and symptoms are usually present in these patients but often missed at ED triage. The diversity of clinical presentations among ED patients with pulmonary TB will likely make it difficult to develop and implement high-yield triage screening criteria. PMID- 11044005 TI - The accuracy of oral predictive and infrared emission detection tympanic thermometers in an emergency department setting. AB - OBJECTIVE: To assess the accuracy of an oral predictive thermometer and an infrared emission detection (IRED) tympanic thermometer in detecting fever in an adult emergency department (ED) population, using an oral glass mercury thermometer as the criterion standard. METHODS: This was a single-center, nonrandomized trial performed in the ED of a metropolitan tertiary referral hospital with a convenience sample of 500 subjects. The temperature of each subject was taken by an oral predictive thermometer, an IRED tympanic thermometer set to "oral" equivalent, and an oral glass mercury thermometer (used as the criterion standard). A fever was defined as a temperature of 37.8 degrees C or higher. The subject's age, sex, triage category, and diagnostic group were also recorded. Sensitivity, specificity, positive and negative likelihood ratios, positive and negative predictive values, and corresponding 95% confidence intervals were calculated. Logistic regression was used to identify predictors of fever. RESULTS: The sensitivities and specificities for detection of fever of the predictive and the IRED tympanic thermometers were similar (sensitivity 85.7%/88.1% and specificity 98.7%/95.8%, respectively). The predictive thermometer had a better positive predictive value (85.7%) compared with the IRED tympanic thermometer (66.1%). The positive and negative likelihood ratios for the predictive oral thermometer were 65 and 0.14, respectively, and for the IRED tympanic thermometer 21 and 0.12, respectively, indicating that the predictive thermometer will "miss" 1 in about 7 fevers and the IRED tympanic thermometer will "miss" 1 in about 8 fevers. CONCLUSIONS: Although quick and convenient, oral predictive and IRED tympanic thermometers give readings that cannot always be relied on in the detection of fever. If we are to continue using electronic thermometers in the ED setting, we need to recognize their limitations and maintain the importance of our clinical judgment. PMID- 11044006 TI - Use of bilevel positive airway pressure in out-of-hospital patients. AB - OBJECTIVE: To evaluate the utility of bilevel positive airway pressure (BiPAP) in the out-of-hospital treatment of patients with presumed congestive heart failure (CHF). METHODS: This was a prospective, sequential, parallel trial in an urban setting served by a single emergency medical services (EMS) system between January 4 and April 15, 1999. A convenience sampling of adults who were transported by rescue units judged to be in CHF by treating emergency medical technicians trained in advanced life support (ALS EMTs) was included. Rescue squads were divided into demographically matched pairs, and one of each was equipped with a BiPAP ventilatory support unit. Bilevel positive airway pressure therapy was added to the existing treatment protocols for eligible study patients. Main outcome measures were out-of-hospital treatment time, oxygen saturation changes, hospitalization length, need for endotracheal intubation, mortality rate, and ease of use of the device by EMS personnel. RESULTS: Sixty two of 71 enrolled patients completed the study. Out-of-hospital treatment times did not differ between groups (31.2 minutes vs 31.4 minutes; p = 0.931). The difference between pre- and post-treatment oxygen saturation levels was greater for the BiPAP group (13.71%) than the control group (6.69%) (p < 0.05). There was no statistical difference between groups in the length of hospital stay [control: 7.63 days, vs BiPAP: 6.33 days, p = 0.48], the intubation rate [control: 7 of 25 (28%) vs BiPAP: 4 of 37 (11%), p = 0.10], or death rate [control: 2 of 24, vs BiPAP: 6 of 37, p = 0.46]. All of the ALS EMTs who used BiPAP thought that it was safe to use, and 97% thought it was easy and appeared to improve patients' dyspnea and respiratory distress. CONCLUSIONS: ALS EMTs can be trained to deliver noninvasive ventilation with BiPAP, find it easy to apply, and believe that it helps relieve dyspnea in patients with suspected CHF. PMID- 11044007 TI - The clinical utility of serologic markers in the evaluation of the acute scrotum. AB - OBJECTIVE: To examine the presence of interleukin-1 (IL-1), interleukin-6 (IL-6), and creatine phosphokinase-MM (CPK-MM) in patients with acute scrotal pain and assess their clinical utility in the diagnosis of testicular torsion (TT) and epididymitis. METHODS: Twenty-five patients with acute scrotal pain were prospectively enrolled over a two-year period. History, physical examination, complete blood count, urinalysis, and scrotal ultrasound were performed. Testicular torsion was confirmed by surgical exploration. Epididymitis was diagnosed using physical examination, scrotal ultrasound, and positive urinalysis. Venous blood was assayed for IL-1, IL-6, and CPK-MM in triplicate during the routine drawing of blood in the emergency department. The data are reported as medians +/- interquartile ranges (IQRs). RESULTS: Twenty-five patients with acute scrotal pain were evaluated; 11 with TT, three with torsion of the appendix testis (TAT), ten with epididymitis, and one with varicocele. One patient had both TT and epididymitis. Interleukin-1 was not detectable in either group. The CPK-MM values between TT and epididymitis were virtually identical at 99.8 and 100 IU/L, respectively. The median value for IL-6 was 1. 03 (IQR = 0.19 to 2.86) vs 20.86 (IQR = 2.14 to 65.50) pg/mL in the torsion and epididymitis groups, respectively. The 97.5% CI for the difference of medians of 19.9 was 0.4 to 65.1, p = 0.02. Using receiver operating characteristic (ROC) curve analysis for IL-6, the area under the curve was 0.82 for torsion and 0.67 for epididymitis. Using a cutoff value of IL-6 >/= 1.41 pg/mL, the positive predictive value of IL-6 in diagnosing epididymitis was 78.6%, with a negative predictive value of 100% for TT. There were no cases of missed TT on follow-up. CONCLUSIONS: This preliminary investigation of serologic markers demonstrates that IL-6 is significantly elevated in epididymitis as compared with TT. Creatine phosphokinase-MM and IL-1 were not found to be of diagnostic utility. The small sample size of this study precludes a definitive conclusion as to the utility of these markers in the emergency department. However, IL-6 may be clinically useful as an additional element in differentiating the causes of acute scrotal pain, and further study is warranted. PMID- 11044008 TI - Massive pneumocephalus following Merocel nasal tamponade for epistaxis. AB - Anterior epistaxis is commonly treated with Merocel nasal tampon insertion in preference to gauze packing. An 89-year-old patient was found to have cerebrospinal fluid rhinorrhea and massive pneumocephalus immediately after removal of a Merocel tampon used for spontaneous anterior epistaxis. She later developed fever and mild confusion, but was well at three-month follow-up. The advantages and complications of Merocel nasal tamponade are briefly reviewed and compared with those of other methods of control of anterior epistaxis. PMID- 11044009 TI - Heeding the call: radiologists in the ED (Emergency Department). PMID- 11044010 TI - Lung cancer detection in the 21st century: potential contributions and challenges of emerging technologies. PMID- 11044011 TI - Value, quality, excellence: our national pastime. PMID- 11044012 TI - 1997 Graduates Speak: initial employment experiences of residency and fellowship graduates. AB - OBJECTIVE: The American College of Radiology sought to detail the initial employment experience of 1997 diagnostic radiology graduates and recent trends. MATERIALS AND METHODS: In early 1998, questionnaires were mailed to all graduates; 65% responded. Results were compared with similar surveys of 1995 and 1996 graduates. RESULTS: Much as in past years, immediately after graduation 4% of residency graduates and 9% of fellowship graduates spent some time working as a locum tenens, working outside radiology, or unemployed, but by 6-12 months after graduation, 2% or fewer were not employed. The median fellowship graduate's salary was 5% greater than in 1996. Twelve percent of fellowship graduates were in non-ownership-track jobs, much the same as in earlier years. Thirty-two percent had one or more of 12 possibly undesirable job characteristics, down from 39% in 1996; 14% not only had, but actually disliked, one or more of these characteristics, much the same as in 1996, but down from 23% in 1995. Multivariate analysis showed that women graduates were more likely than men to have had serious employment difficulties in the immediate postgraduation months; and that graduates having a spouse who also had to find a job in the same area were more likely than others to be in a putatively "seriously undesirable" location or in "holding pattern" employment while looking for something better or more permanent. CONCLUSION: Unemployment remained very low. Some other indicators of the employment market showed improvement. Factors expected to affect employment outcomes-for example, training program quality or having major non spouse-related restrictions on job location-had surprisingly little effect. PMID- 11044013 TI - Interpretation of Emergency Department radiographs: a comparison of emergency medicine physicians with radiologists, residents with faculty, and film with digital display. AB - OBJECTIVE: We determined the relative value of teleradiology and radiology resident coverage of the emergency department by measuring and comparing the effects of physician specialty, training level, and image display method on accuracy of radiograph interpretation. MATERIALS AND METHODS: A sample of four faculty emergency medicine physicians, four emergency medicine residents, four faculty radiologists, and four radiology residents participated in our study. Each physician interpreted 120 radiographs, approximately half containing a clinically important index finding. Radiographs were interpreted using the original films and high-resolution digital monitors. Accuracy of radiograph interpretation was measured as the area under the physicians' receiver operating characteristic (ROC) curves. RESULTS: The area under the ROC curve was 0.15 (95% confidence interval [CI], 0.10-0.20) greater for radiologists than for emergency medicine physicians, 0.07 (95% CI, 0.02-0.12) greater for faculty than for residents, and 0.07 (95% CI, 0.02-0.12) greater for films than for video monitors. Using these results, we estimated that teleradiology coverage by faculty radiologists would add 0.09 (95% CI, 0.03-0.15) to the area under the ROC curve for radiograph interpretation by emergency medicine faculty alone, and radiology resident coverage would add 0.08 (95% CI, 0.02-0.14) to this area. CONCLUSION: We observed significant differences between the interpretation of radiographs on film and on digital monitors. However, we observed differences of equal or greater magnitude associated with the training level and physician specialty of each observer. In evaluating teleradiology services, observer characteristics must be considered in addition to the quality of image display. PMID- 11044014 TI - Language of the radiology report: primer for residents and wayward radiologists. PMID- 11044015 TI - The deep pocket. PMID- 11044016 TI - Automated teaching file and slide database for digital images. AB - OBJECTIVE: We developed an easy-to-use method for creating a searchable digital teaching file of CT and MR images. CONCLUSION: We describe a method of creating a digital archive of interesting cases that is easy to implement and works on a commercially available workstation. A remote daemon polls for images transferred to the product film tool. It creates a tagged image file format (TIFF) digital archive of these images on any platform supporting file transfer protocol (FTP), operates in the background, and automatically generates a searchable index of case information in the database. PMID- 11044017 TI - Creation of radiofrequency lesions in a porcine model: correlation with sonography, CT, and histopathology. AB - OBJECTIVE: We studied the correlation between sonographic and CT appearances of radiofrequency thermal lesions created in porcine liver and histopathologic findings to evaluate the accuracy of these techniques in revealing the extent of tissue necrosis. MATERIALS AND METHODS: We used sonographic guidance and a 2.0-cm diameter, eight-prong retractable radiofrequency electrode to view 12 hepatic lesions that were created in five pigs. Biphasic helical CT was performed 12-48 hr after ablation. The animals were sacrificed immediately after CT, and their livers were histopathologically examined. The maximum lesion size in the long and short axes as measured on CT and sonography was then correlated with the histopathologically determined lesion size. RESULTS: On sonography, lesions changed rapidly within 5 min after the termination of ablation. An early echogenic cloud became peripherally hypoechoic with a variable thin echogenic rim. Early (0-2 min after ablation) sonograms led to an underestimation of true lesion sizes on histopathology (r = 0.3-0.49; p < 0.05). Delayed (2-5 min after ablation) sonograms also led to an underestimation of true lesion size (r = 0.5 0.62; p < 0.05); however, lesions were larger and better demarcated. Biphasic contrast-enhanced helical CT revealed avascular lesions surrounded by hyperemic rims that closely correlated with true pathologic lesions size (r = 0.93-0.95; p < 0. 05). Lesions with hyperemic rims that were measured on CT led to overestimations of true lesion size. CONCLUSION: Sonography led to underestimations of the true size of ablated lesions within the first 5 min after creation; however, delayed images provided better results. The avascular lesion measured on contrast-enhanced helical CT closely correlated with the size of ablated tissue; therefore, contrast-enhanced CT is preferred for serially monitoring the effect of radiofrequency ablation. PMID- 11044018 TI - Localization of hepatocellular carcinoma in the hepatic dome before tumor ablation: using a system that includes a hookwire and suture. PMID- 11044019 TI - Radiofrequency ablation of spinal tumors: temperature distribution in the spinal canal. PMID- 11044021 TI - Role of the interventional radiologist in treating obstetric-gynecologic pathology. PMID- 11044020 TI - Uterine artery embolization in the primary treatment of uterine leiomyomas: technical features and prospective follow-up with clinical and sonographic examinations in 58 patients. AB - OBJECTIVE: The purpose of this study was to analyze the technical features of uterine artery embolization and to evaluate the effectiveness of this method as the primary treatment of uterine leiomyomas in a series of 58 patients monitored by clinical and sonographic examinations. SUBJECTS AND METHODS: Fifty-eight women (age range, 33-65 years; mean age, 44.5 years) with symptoms caused by uterine leiomyomas (abnormal bleeding, bulk-related symptoms, pelvic pain) were included in this prospective study. We performed embolization with a single catheter using the single-femoral artery approach, injection of particles (150-250 mm), and an absorbable gelatin sponge. Postprocedural pain was assessed using a visual analog scale. Systematic follow-up included clinical and sonographic examinations at 3 months for 58 patients, at 6 months for 46 patients, at 1 year for 27 patients, and at 2 years for seven patients. RESULTS: Embolization was performed without problems in 84% of the patients. Post-procedural pain control was excellent in 90% of the patients. In most patients, symptoms were improved or had resolved at 3 months (90%), 6 months (92%), and 1 year (93%), and all patients were symptom free at 2 years. Clinical failure of treatment occurred in only two patients (3%). Progressive reduction in leiomyoma size was revealed during sonographic follow-up, and new leiomyomas were seen in one patient at 2 years. CONCLUSION: Uterine artery embolization is an endovascular method for the treatment of uterine leiomyomas that is clinically effective in most patients and that induces a progressive reduction in the size of the largest leiomyomas. PMID- 11044022 TI - Inter- and intraobserver variability of CT measurements obtained after endovascular repair of abdominal aortic aneurysms. AB - OBJECTIVE: Important decisions are made on the basis of CT angiographic measurements of aneurysm size obtained after endovascular abdominal aortic aneurysm repair; however, little is known about the variability of these measurements. We evaluated the variability of CT angiographic measurements of aneurysm size obtained after endovascular abdominal aortic aneurysm repair. MATERIALS AND METHODS: Thirty CT angiographic data sets were randomly chosen from 91 sets, including preoperative, postoperative, and follow-up CT images. All images were obtained according to a standardized acquisition protocol. On a workstation, three parameters were measured: maximum aneurysm diameter, maximum aneurysm cross-sectional area, and aneurysm volume. All data sets were measured twice by two investigators in a random order. The difference of each pair of measurements was plotted against the mean value. The mean difference and its standard deviation were calculated with a repeatability coefficient. RESULTS: The intraobserver repeatability coefficient for observer 1 was 3.8 mm for diameter, 201.7 mm(2) for cross-sectional area, and 5.6 mL for volume. For observer 2, these figures were 3.0 mm, 219.0 mm(2), and 8.1 mL, respectively. The interobserver repeatability coefficients were 3.9 mm, 236.2 mm(2), and 10.3 mL. CONCLUSION: Determination of the repeatability coefficient of aneurysm size measurements obtained after endovascular abdominal aortic aneurysm repair provides a good description of precision. PMID- 11044023 TI - Percutaneous stent treatment for arterial occlusion caused by retroperitoneal fibrosis. PMID- 11044024 TI - Transient aortic thrombus in a sickle-cell patient with chest pain: CT and MR findings. PMID- 11044025 TI - Intimal sarcoma of the aorta. PMID- 11044026 TI - Dynamic contrast-enhanced MR angiography from the distal aorta to the ankle joint with a step-by-step technique. AB - OBJECTIVE: The aim of this study was to visualize the arteries from the distal aorta to the ankle joint and to determine the accuracy of MR angiography for detecting stenoses and occlusions. SUBJECTS AND METHODS: Twenty-four patients with peripheral arterial occlusive disease underwent digital subtraction angiography and were examined on a 1.5-T MR scanner. The transit time for contrast material was determined with a test bolus injection. A T1-weighted three dimensional gradient-echo sequence with short TR and TE was used for a dynamic measurement at the level of the iliac arteries, the upper leg, and the lower leg arteries. For each level a single dose of gadolinium was injected into an antecubital vein with an MR power injector. Maximal-intensity-projection reconstructions were calculated after subtraction of the first measurement at each level. Two experienced MR radiologists who were unaware of the digital subtraction angiography results interactively evaluated both the MIP reconstructions and the single slices on a workstation, first independently and then in a consensus interpretation. RESULTS: With digital subtraction angiography, 80 hemodynamically significant stenoses and 39 occlusions were detected. For the stenoses and occlusions, a sensitivity of 100% was found for MR angiography. The specificity for the assessment of stenoses and occlusions was 98% and 94%, respectively, for the iliac arteries; 98% and 94%, respectively, for the upper leg arteries; and 94% and 95%, respectively, for the lower leg arteries. Most false-positive findings of occlusion were due to metal stents present in the iliac (n = 3) and upper leg (n = 4) arteries. CONCLUSION: The MR imaging technique that we used revealed the arteries from the distal aorta to the ankle and proved to be reliable at showing arterial stenoses and occlusions. PMID- 11044027 TI - Distribution of thrombi in acute lower extremity deep venous thrombosis: implications for sonography and CT and MR venography. AB - OBJECTIVE: Our objective was to determine the typical distribution of thrombi in acute lower extremity deep venous thrombosis as a means of evaluating the validity of imaging techniques that only include the common femoral and popliteal veins, but not the superficial femoral vein. MATERIALS AND METHODS: The results of 2704 lower extremity venous sonograms, obtained in 2026 consecutive patients over a 4-year interval, were reviewed retrospectively. The distribution of acute deep venous thromboses across various lower extremity venous segments was analyzed for this population, which consisted of both symptomatic and asymptomatic patients. RESULTS: Of 2704 lower extremities studied with duplex sonography, acute deep venous thrombosis was identified in 269 (9.9%). Of these 269 cases, acute deep venous thrombosis was isolated to the superficial femoral vein in 60 (22.3%). The remaining 209 cases (77.7%) showed thrombus that extended into the common femoral or popliteal veins (or both). CONCLUSION: An abbreviated imaging study that evaluates only the common femoral and popliteal veins would fail to identify more than 20% of lower extremity acute deep venous thromboses in a population like ours. Although evaluation of the superficial femoral vein requires additional time and resources, evaluation of this segment may prevent a substantial number of thrombi from being missed. PMID- 11044028 TI - Percutaneous translumbar placement of a Hickman catheter into the azygous vein. PMID- 11044029 TI - Osteochondritis dissecans of the tibial plafond: imaging characteristics and a review of the literature. AB - OBJECTIVE; Osteochondritis dissecans of the talar articular surface of the ankle joint has been well described. We report the imaging characteristics of osteochondritis dissecans of the tibial articular surface (tibial plafond). CONCLUSION: Osteochondritis dissecans of the tibial plafond is a rare condition that may not be detectable on radiography. Its radiologic findings are similar to those of osteochondritis dissecans located elsewhere in the body. PMID- 11044030 TI - Cervical radiography for trauma patients: a time-effective technique? AB - OBJECTIVE: The purpose of this study was to determine the time necessary to perform a six-view radiographic examination of the cervical vertebral column of trauma victims. In addition we compared the added examination times for 30 patients who underwent an additional helical CT examination of the cervical region immediately after cranial CT. CONCLUSION: Cervical radiography is a time consuming procedure, which is a concern for trauma surgeons. A more efficient way for cervical evaluation of trauma patients needs to be adopted. Evidence now exists in the literature to suggest that helical CT can serve that purpose. PMID- 11044031 TI - An illustrated tutorial of musculoskeletal sonography: part 3, lower extremity. PMID- 11044032 TI - Transoral approach to cervical vertebroplasty for multiple myeloma. PMID- 11044034 TI - Advanced imaging of the diabetic foot and its complications PMID- 11044033 TI - Schnitzler's syndrome: an unusual cause of bone pain with suggestive imaging features. PMID- 11044035 TI - Automatic detection and quantification of ground-glass opacities on high resolution CT using multiple neural networks: comparison with a density mask. AB - OBJECTIVE: We compared multiple neural networks with a density mask for the automatic detection and quantification of ground-glass opacities on high resolution CT under clinical conditions. SUBJECTS AND METHODS: Eighty-four patients (54 men and 30 women; age range, 18-82 years; mean age, 49 years) with a total of 99 consecutive high-resolution CT scans were enrolled in the study. The neural network was designed to detect ground-glass opacities with high sensitivity and to omit air-tissue interfaces to increase specificity. The results of the neural network were compared with those of a density mask (thresholds, -750/-300 H), with a radiologist serving as the gold standard. RESULTS: The neural network classified 6% of the total lung area as ground-glass opacities. The density mask failed to detect 1.3%, and this percentage represented the increase in sensitivity that was achieved by the neural network. The density mask identified another 17.3% of the total lung area to be ground glass opacities that were not detected by the neural network. This area represented the increase in specificity achieved by the neural network. Related to the extent of the ground-glass opacities as classified by the radiologist, the neural network (density mask) reached a sensitivity of 99% (89%), specificity of 83% (55%), positive predictive value of 78% (18%), negative predictive value of 99% (98%), and accuracy of 89% (58%). CONCLUSION: Automatic segmentation and quantification of ground-glass opacities on high-resolution CT by a neural network are sufficiently accurate to be implemented for the preinterpretation of images in a clinical environment; it is superior to a double-threshold density mask. PMID- 11044036 TI - Lymphomatoid granulomatosis: radiologic features and pathologic correlations. AB - OBJECTIVE: The purpose of this paper is to define the radiologic features of lymphomatoid granulomatosis and correlate them with histopathologic features. CONCLUSION: Lymphomatoid granulomatosis shows characteristic CT features such as peribronchovascular distribution of nodules, coarse irregular opacities, small thin-walled cysts, and conglomerating small nodules. Large masses and occlusion of large vessels also occur. Histopathologic examination shows the nodules are caused by intravascular and perivascular infiltration by atypical lymphoid cells. PMID- 11044037 TI - Primary liposarcoma of the mediastinum. PMID- 11044039 TI - Case-based reasoning computer algorithm that uses mammographic findings for breast biopsy decisions. AB - OBJECTIVE: We present case-based reasoning computer software developed from mammographic findings to provide support for the clinical decision to perform biopsy of the breast. SUBJECTS AND METHODS: The case-based reasoning system is designed to support the decision to perform biopsy in those patients who have suspicious findings on diagnostic mammography. Currently, between 66% and 90% of biopsies are performed on benign lesions. Our system is designed to help decrease the number of benign biopsies without missing malignancies. Clinicians interpret the mammograms using a standard reporting lexicon. The case-based reasoning system compares these findings with a database of cases with known outcomes (from biopsy) and returns the fraction of similar cases that were malignant. This malignancy fraction is an intuitive response that the clinician can then consider when making the decision regarding biopsy. RESULTS: The system was evaluated using a round-robin sampling scheme and performed with an area under the receiver operating characteristic curve of 0.83, comparable with the performance of a neural network model. If only the cases returning a malignancy fraction of greater than a threshold of 0.10 are sent to biopsy, no malignancies would be missed, and the number of benign biopsies would be decreased by 25%. At a threshold of 0.21, 98%, of the malignancies would be biopsied, and the number of benign biopsies would be decreased by 41%. CONCLUSION: This preliminary investigation indicates that the case-based reasoning approach to computer-aided diagnosis has the potential to improve the accuracy of breast cancer diagnosis on mammography. PMID- 11044038 TI - Atypical ductal hyperplasia and ductal carcinoma in situ as revealed by large core needle breast biopsy: results of surgical excision. AB - OBJECTIVE: This investigation compares the frequency of histologic underestimation of breast carcinoma that occurs when a large-core needle biopsy reveals atypical ductal hyperplasia or ductal carcinoma in situ with the automated 14-gauge needle, the 14-gauge directional vacuum-assisted biopsy device, and the 11-gauge directional vacuum-assisted biopsy device. SUBJECTS AND METHODS: Evaluation of 428 large-core needle biopsies yielding atypical ductal hyperplasia (139 lesions) or ductal carcinoma in situ (289 lesions) was performed. The results of subsequent surgical excision were retrospectively compared with the needle biopsy results. RESULTS: For lesions initially diagnosed as ductal carcinoma in situ, underestimation of invasive ductal carcinoma was significantly less frequent using the 11-gauge directional vacuum-assisted biopsy device when compared with the automated 14-gauge needle (10% versus 21%, p < 0.05) but was not significantly less frequent when compared with the 14-gauge directional vacuum-assisted device (10% versus 17%, p > 0.1). For lesions diagnosed initially as atypical ductal hyperplasia, underestimation of ductal carcinoma in situ and invasive ductal carcinoma was significantly less frequent using the 11-gauge directional vacuum-assisted biopsy device compared with the 14 gauge directional vacuum-assisted device (19% versus 39%, p = 0. 025) and with the automated 14-gauge needle (19% versus 44%, p = 0. 01). CONCLUSION: The frequency of histologic underestimation of breast carcinoma in lesions initially diagnosed as atypical ductal hyperplasia or ductal carcinoma in situ using large core needle biopsy is substantially lower with the 11-gauge directional vacuum assisted device than with the automated 14-gauge needle and with the 14-gauge directional vacuum-assisted device. PMID- 11044040 TI - Sonographically guided metallic clip placement after core needle biopsy of the breast. PMID- 11044041 TI - Identification of feeding arteries to establish the intra- or extraparotid location of jugulodigastric nodules: value of color Doppler sonography. AB - OBJECTIVE: The objective of our study was to determine the value of color Doppler imaging as an adjunct to gray-scale sonography to reveal the intra- or extraparotid origin of jugulodigastric nodules of uncertain location. SUBJECTS AND METHODS: Forty nodules in the jugulodigastric area from 38 patients were imaged with gray-scale, color, and power Doppler sonography. Nodules receiving vessels from salivary glands were assumed to be intraglandular; those accepting vessels from paraparotid spaces were considered to arise from outside the gland. Imaging results were correlated with biopsy and surgical findings. RESULTS: In 38 of 40 cases, color and power Doppler sonography displayed discrete feeding arteries leading to the nodules: 25 nodules had one supplying artery, nine received two arteries, and four had three or more arteries. Intraparotid nodules received vessels from the gland in 20 cases. In two cases, the source of vessels was indeterminate. In two malignant tumors, multiple arteries derived from both the parotid and the neck spaces. All 14 extraparotid nodules received the arterial supply from paraparotid spaces. Color Doppler sonography enabled prediction of the intraglandular location of the nodules in 91% of cases and the extraglandular location in 87.5% of cases. The correct diagnosis was achieved in 34 of 40 nodules, with a global accuracy of 85%. CONCLUSION: Color Doppler sonography can help to assess the intra- or extraparotid location of jugulodigastric nodules. In practice, this technique can support the diagnosis when gray-scale sonography raises doubts about the origin of a jugulodigastric nodule. PMID- 11044042 TI - Thick-section reformatting of thinly collimated helical CT for reduction of skull base-related artifacts. AB - OBJECTIVE: Our purpose was to evaluate thick-section reformatted helical CT of the brain base as a technique for reducing skull base-related artifacts and to compare it with conventional CT. MATERIALS AND METHODS: Twenty-three patients with suspected intracranial abnormalities related to the brain base, as determined either by clinical examination or at the time of imaging, were evaluated with contrast-enhanced conventional CT of the brain (5-mm collimation, 140 kVp, 170 mA, 2-sec rotation time) and reformatted helical CT (1-mm collimation, 1.5 pitch, 120 kVp, 220 mA). Helical sections were reformatted to a thickness of 5 mm by a volume-averaging algorithm using a computer workstation. Three observers retrospectively and blindly reviewed the images and qualitatively scored artifacts at the foramen magnum, middle cranial fossa, anterior cranial fossa, interpetrous region, and internal occipital protuberance. Image graininess and observer confidence were also scored. Paired statistical analyses using score differences in each patient were possible. RESULTS: Reformatted helical CT reduced skull base-related artifacts across all five anatomic regions (p < 0.05). The foramen magnum showed the greatest reduction in artifacts and the anterior cranial fossa the least. Image graininess was increased on reformatted CT compared with conventional CT (p < 0.05), but observer confidence remained higher for reformatted CT (p < 0.05). Total additional scan time was 3.15 +/- 0.38 min with 5.3 +/- 1.2 min required for reformatting. CONCLUSION: Reformatted CT significantly decreases skull base-related artifacts in the brain, improving confidence in evaluation of the brain base and adding an average of only 8.45 +/- 1.6 min of scanning and processing time to each examination. PMID- 11044043 TI - Primary basilar artery stenting: immediate and long-term results in one patient. PMID- 11044044 TI - Computerized tomography (CT) in acute head trauma PMID- 11044045 TI - Internet-based interactive teaching file for neuroradiology. AB - OBJECTIVE: Our goal was to implement an interactive neuroradiologic teaching file that can be accessed on the Internet and easily expanded to include radiologic, clinical, and pathologic correlation. CONCLUSION: Our growing interactive neuroradiologic teaching file is available on the Internet. It provides an easily accessed database of interesting cases to aid in the study or analysis of difficult cases. PMID- 11044046 TI - The egyptian spice market, istanbul, 1999 PMID- 11044047 TI - T2-Weighted fast MR imaging with true FISP versus HASTE: comparative efficacy in the evaluation of normal fetal brain maturation. AB - OBJECTIVE: This study compares the relative efficacy of two fast T2-weighted MR imaging techniques-fast imaging with steady-state free precession (true FISP) and half-Fourier acquisition single-shot turbo spin-echo (HASTE)-in the evaluation of the normal fetal brain maturation during the second and third trimesters of gestation. SUBJECTS AND METHODS: The brain maturation of 10 normal nonsedated fetuses (5 during the second trimester and 6 during the third trimester of gestation [1 fetus underwent 2 examinations]) was examined by both techniques using a Vision+ 1.5-T MR system. We specifically looked for developing events, including white matter myelination, neuronal migration, and cortical sulcation. Image quality was graded according to the presence or absence of undesirable blurring. RESULTS: The specific absorption rate was lower for true FISP than for HASTE by a factor of 3 at equivalent imaging conditions. HASTE and true FISP provide comparable image quality in the second trimester when myelination of the cerebrum has not begun. Neuronal migration could be recognized as hypodense bands on both sequences during the second trimester. Myelination beginning at the third trimester was better delineated with true FISP than with HASTE because of point spread function-related blurring effects inherent in HASTE that hampered visualization of short-T2 structures. Cortical sulcation was well delineated by both sequences. CONCLUSION: With relatively superior image quality and significantly lower radiofrequency absorption than HASTE, true FISP is a safer and more effective alternative in the prenatal evaluation of normal fetal brain. PMID- 11044048 TI - Helical CT angiography and three-dimensional reconstruction of total anomalous pulmonary venous connections in neonates and infants. AB - OBJECTIVE: The objective of this study was to investigate the usefulness of helical CT angiography in the evaluation of total anomalous pulmonary venous connections. MATERIALS AND METHODS: Fourteen patients with total anomalous pulmonary venous connections underwent helical CT angiography and subsequent three-dimensional (3D) reconstruction. They ranged in age from 3 days to 8 months (median age, 2.3 months) and in weight from 2.3 to 7.1 kg (median weight, 4.3 kg). The types of total anomalous pulmonary venous connections and the number of pulmonary veins were evaluated on axial and 3D images. Qualitative evaluations were performed for extent of pulmonary vascular enhancement and contrast- or motion-induced artifacts. RESULTS: In all patients, helical CT angiography correctly depicted total anomalous pulmonary venous connections. Seven cases were the supracardiac type, four cases were the cardiac type, one case was the infracardiac type, and two cases were the mixed type. The detection rate of the pulmonary vein in 3D reconstruction images (95-98%) was slightly lower than that of the pulmonary vein in the axial images (100%), but the difference between axial and 3D reconstruction images was not statistically significant (p > 0.1). No statistically significant differences were noted among 3D reconstruction images in the detection rates of the pulmonary vein (p > 0.1). The extent of contrast enhancement of the pulmonary vein was good or excellent in all patients. In five patients, there were contrast-induced artifacts that made some surrounding vascular distortion but did not interfere with the pulmonary vein analysis, except in one patient. Motion-induced artifacts were observed in nine patients. One of them had an obstacle in pulmonary vein analysis. CONCLUSION: The combination of axial and 3D images in helical CT angiography is helpful in the assessment of a total anomalous pulmonary venous connection containing the individual pulmonary vein, and this combination can be a good diagnostic tool in preoperative evaluation of neonates and infants with a total anomalous pulmonary venous connection. PMID- 11044049 TI - Accuracy of noncompressive sonography of children with appendicitis according to the potential positions of the appendix. AB - OBJECTIVE: This study evaluates noncompressive sonography of appendicitis in children according to the potential positions of the appendix. SUBJECTS AND METHODS: We evaluated 425 consecutive boys and girls clinically suspected of having appendicitis. Noncompressive sonography used a 5.0-MHz curved-array transducer to assess deep layers. We systematically investigated the possible positions of the appendix by evaluating the right retrocecal region through the flank, the pelvis through the suprapubic region with the urinary bladder full, and the right lower quadrant. Whenever the appendix was not visualized, graded compression examination was performed with 5.0-MHz curved-array and 7.5-MHz linear transducers, and the remainder of the abdomen was thoroughly examined with 5.0- and 3.75-MHz curved-array transducers. Sonographic findings were correlated with pathology findings after surgery in 212 cases and with clinical follow-up in 213. RESULTS: Of the 425 patients with abdominal pain, 199 had surgically confirmed appendicitis; noncompressive examination accurately identified 135 (31.7%) of these 425 cases, and the combined noncompressive and compressive examinations diagnosed 196 cases. The combined noncompressive and compressive techniques had a sensitivity of 98.5% (95% confidence interval, 96.8-100), specificity of 98.2% (96.5-99.9), positive and negative predictive values of 98.0% (99.9) and 98.7% (97.2-100), respectively. CONCLUSION: The noncompressive technique is a valuable tool in sonographic investigation of appendicitis. PMID- 11044050 TI - CT-Guided transgluteal drainage of deep pelvic abscesses in children: selective use as an alternative to transrectal drainage. AB - OBJECTIVE: The transgluteal approach to abscess drainage through the greater sciatic foramen has been described in adults, but this route has not been as extensively studied in children. We performed CT-guided transgluteal percutaneous abscess drainage in seven children and assessed the results of drainage and catheter tolerance. CONCLUSION: Transgluteal catheters are well tolerated by children, and the transgluteal route is an effective approach to selected pelvic abscesses in children. PMID- 11044051 TI - Angiographic findings of persistent primitive hepatic venous plexus with underdevelopment of the infrahepatic inferior vena cava in pediatric patients. AB - OBJECTIVE: We describe the angiographic diagnosis and significance of persistence of the primitive hepatic venous plexus with underdevelopment of the infrahepatic inferior vena cava. CONCLUSION: We recommend that inferior venacavography be performed in routine assessment before surgery of patients with azygos or hemiazygos continuation of the inferior vena cava, in whom redirection of systemic venous blood to the pulmonary artery is contemplated. PMID- 11044052 TI - Three-dimensional CT of congenital esophageal atresia and distal tracheoesophageal fistula in neonates: preliminary results. AB - OBJECTIVE: Radiography was traditionally used in the preoperative treatment of neonates with tracheoesophageal atresia and tracheoesophageal fistula. The aim of this study was to assess the potential use of three-dimensional CT in the evaluation of this complex congenital malformation. CONCLUSION: Three-dimensional CT coupled with reformations in the three orthogonal planes may have a complementary diagnostic role in congenital esophageal atresia. PMID- 11044053 TI - Prenatal diagnosis of giant congenital pancreatic cyst of a neonate. PMID- 11044054 TI - Delayed enhanced CT of lipid-poor adrenal adenomas. AB - OBJECTIVE: Although representing a minority of adrenal adenomas, the lipid-poor variety cannot be accurately identified on unenhanced CT or chemical shift MR imaging. We compared the delayed contrast-enhanced CT features of lipid-poor adenomas with those of lipid-rich adenomas and of adrenal nonadenomas to determine whether there were differences in the washout features between these groups of lesions. SUBJECTS AND METHODS: Eighteen proven lipid-poor adenomas, 56 lipid-rich adenomas, and 40 adrenal nonadenomas underwent CT before, immediately after, and 15 min delay after IV contrast injection. Region-of-interest measurements were made of all adrenal lesions at the three time points. The degree of enhancement, enhancement washout, percentage enhancement washout, and relative percentage enhancement washout were calculated for each adrenal mass. Pooled data were analyzed statistically. Optimal threshold values for diagnosing adrenal adenomas were also determined. RESULTS: The mean CT attenuation of lipid poor adenomas was significantly higher than that of lipid-rich adenomas at all three phases but not significantly different from that of nonadenomas. The mean percentage enhancement washout on images obtained 15 min after administration of contrast material was similar for lipid-rich and lipid-poor adenomas but was significantly higher than that of nonadenomas. The mean relative percentage enhancement washout was significantly different among all three groups. CONCLUSION: Lipid-poor adenomas cannot be differentiated from adrenal nonadenomas on the basis of a single mean attenuation value. However, lipid-poor adrenal adenomas show enhancement and enhancement washout features nearly identical to lipid-rich adenomas and can be distinguished from nonadenomas on the basis of a percentage washout threshold value of 60% and a relative percentage washout of 40%. PMID- 11044055 TI - Focal posttransplantation lymphoproliferative disorder at the renal allograft hilum. AB - OBJECTIVE: This report describes the imaging characteristics of focal posttransplantation lymphoproliferative disorder. CONCLUSION: Posttransplantation lymphoproliferative disorder may be limited to the allograft. A focal complex mass in the renal allograft hilum surrounding the main renal blood vessels is a common finding and can be visualized with sonography. MR imaging can help increase diagnostic confidence. PMID- 11044056 TI - Ovarian carcinoma in patients with endometriosis: MR imaging findings. AB - OBJECTIVE: Various types of malignancy can develop in patients with endometriosis. Enhancing mural nodules have been reported as an imaging characteristic of malignant transformations. We evaluated contrast-enhanced MR imaging to determine the optimum sequence to reveal mural nodules and other characteristics of malignant transformations. MATERIALS AND METHODS: We examined 10 patients with pathologically proven ovarian adenocarcinoma in endometriosis and 10 patients (the control group) with ovarian endometrioma suggestive of malignant transformation on the basis of sonographic findings. We analyzed the size and nature of the endometriomas in each patient. We compared four types of contrast-enhanced MR imaging to determine which sequence best revealed mural nodules. RESULTS: In the malignant and control groups, 80% of the cysts with findings suggestive of malignant transformation showed unilateral disease or larger endometrial cysts on the suggestive side than on the contralateral side. High signal intensity on T1-weighted images and low signal intensity on T2 weighted images relative to the myometrium were observed only in two of 10 malignant endometrial cysts and in all control cysts. All malignant endometriomas had small mural nodules with low signal intensity on T1-weighted contrast enhanced images. Only three benign endometriomas had mural nodules and none of them enhanced. The enhancement of mural nodules was easily seen on dynamic subtraction imaging. CONCLUSION: On the basis of our findings, endometrial cysts with malignant transformation rarely show low signal intensity on T2-weighted images and usually have enhancing mural nodules. Because the enhancement of mural nodules is often difficult to evaluate on conventional T1-weighted images, dynamic subtraction imaging can be valuable. PMID- 11044057 TI - CT findings in patients with esophagitis. AB - OBJECTIVE: The purpose of our study was to determine the CT findings in patients with esophagitis. MATERIALS AND METHODS: A search of medical and radiology files revealed 29 patients with esophagitis in whom thoracic CT was performed within 1 month of the diagnosis. The CT scans were reviewed retrospectively for esophageal wall thickening, a target sign, or other abnormalities. The thickness of the esophageal wall was also measured on CT in these 29 patients and compared with a control group of 85 patients. RESULTS: We found that 16 patients (55%) with esophagitis had abnormal findings on CT, including a thickened esophageal wall (using 5 mm as the threshold for wall thickening) in all 16 (55%) and a target sign in five (17%). The overall mean esophageal wall thickness was 4.7 mm (standard deviation [SD], 2 mm; 95% confidence interval [CI], 0.7-8.7 mm) in patients with esophagitis versus a mean wall thickness of 2.9 mm (SD, 0.8 mm; 95% CI, 1.3-4.5 mm) in controls (p <.001). Using the same 5-mm threshold for wall thickening, we found that only three (4%) of 85 controls had a thickened esophageal wall on CT. CONCLUSION: The majority of patients with esophagitis had abnormalities on CT, including a thickened esophageal wall (> or =5 mm) in 55% and a target sign in 17%. Although barium studies and endoscopy are more sensitive modalities for detecting this condition, the CT finding of a relatively long segment of circumferential esophageal wall thickening, with or without a target sign, should suggest the diagnosis of esophagitis in the proper clinical setting. PMID- 11044058 TI - Detection of pharyngeal perforation: comparison of aqueous and barium-containing contrast agents. AB - OBJECTIVE: We sought to assess the value of aqueous and barium-containing contrast agents in the detection of pharyngeal perforation. SUBJECTS AND METHODS: Visual and objective in vitro comparisons of an iodinated aqueous contrast agent, a 50% weight/volume barium suspension, and a 100% weight/volume barium suspension were performed. Moreover, to exclude pharyngeal perforation after surgery, we prospectively examined 109 patients by pharyngography, using the aqueous contrast agent and the 100% weight/volume barium suspension. All patients with a pharyngeal perforation were followed up clinically to exclude complications due to barium application. RESULTS: As opposed to the 100% weight/volume barium suspension, in vitro comparison between the aqueous contrast agent and the 50% weight/volume barium suspension yielded no substantial differences. Seventeen perforations could be detected with the aqueous contrast agent. Although 10 of 17 perforations could be slightly better visualized with the 100% weight/volume barium suspension, two perforations were missed with this agent. Five perforations were equally well detected with both. CONCLUSION: Because of a higher radiopacity, 100% weight/volume barium suspensions may more sharply delineate perforations. However, in contrast to aqueous contrast media, narrow pharyngeal perforations can be missed. Thus, the use of a 100% weight/volume barium suspension does not improve the detection of pharyngeal perforation. PMID- 11044059 TI - CT of patients with right-sided colon cancer and distal ileal thickening. AB - OBJECTIVE: The purpose of this study was to analyze the incidence and primary causes of distal ileal wall thickening in 131 patients with right-sided colon cancer. SUBJECTS AND METHODS: During a 2-year period, 131 patients underwent surgical resection for right-sided colon cancer. Of these patients, we analyzed 13 who had distal ileal wall thickening on CT before surgery and also had the cause determined at pathology. CT findings were analyzed with regard to the morphologic features of colonic tumors, bowel wall involvement patterns of the distal ileum, and changes in the pericolic space. RESULTS: Distal ileal wall thickening occurred in 13 (10%) of the 131 patients who had right-sided colon cancer. Three patients had polypoid colon cancer, whereas the other 10 had infiltrative colon cancer. The mean thickness of the involved colonic segments was 1.6 cm (range, 1.0-2.2 cm) with a mean length of 5.2 cm (range, 2.5-10.0 cm). Pericolic infiltration was mild in six patients and moderate in four patients. The mean length and thickness of the affected ileal segments were 3.2 cm (range, 1.5-6.0 cm) and 1.1 cm (range, 0.7-2.0 cm), respectively. On histopathologic examination, neoplastic processes involved the distal ileum in nine (69%) of the 13 patients. This involvement was caused by either direct tumor invasion in seven patients or lymphatic spread in two. In four patients (31%), nonneoplastic processes with edema and congestion involved the distal ileum. CONCLUSION: The distal ileum may be abnormally thickened in about 10% of patients with right sided colon cancer; this thickening results from tumor extension (69%) or a nontumorous process (31%). PMID- 11044060 TI - Giant hyperplastic polyps in the stomach: radiographic findings in seven patients. AB - OBJECTIVE: We reassessed the radiographic findings of giant hyperplastic polyps in the stomach on double-contrast upper gastrointestinal examinations in seven patients. CONCLUSION: Giant hyperplastic polyps in the stomach may be manifested by distinctive findings on double-contrast barium studies, appearing as polypoid lesions with multiple lobulated components that form a conglomerate mass. Nevertheless, endoscopy and biopsy are required to rule out a polypoid carcinoma as the cause of these findings. PMID- 11044061 TI - Feasibility of instructing radiology technologists in the performance of gastrointestinal fluoroscopy. AB - OBJECTIVE: We sought to determine if dedicated gastrointestinal technologists could be trained to properly perform esophagography and double-contrast barium enema examinations. SUBJECTS AND METHODS: Ninety-four patients undergoing double contrast barium enema examinations and 123 patients undergoing esophagographic examinations were included in the study. The study was conducted over a 4-month period, with examinations performed by eight gastrointestinal technologists, 10 radiology residents, and four staff radiologists. Four random lists were generated for each set of examinations. Each staff gastrointestinal radiologist, who was unaware of who had performed the examination, independently scored the representative radiographs. RESULTS: For the double-contrast barium enema examinations, no statistically significant differences were found between the technologists and residents for amount of barium used, degree of distention, cecal opacification, and quality of spot radiographs. The technologist-performed examinations had a statistically significant lower mean fluoroscopy time (3.2 min, compared with 4.0 min for staff radiologists and 5.7 min for residents). For the esophagrams, no statistically significant differences between technologists and residents were found for single-contrast esophagrams; radiographs of the gastric cardia; assessment of motility, reflux, and transit of a solid bolus; and fluoroscopy time. Double-contrast esophagrams obtained by technologists received a better mean score than did those of the residents. CONCLUSION: Radiology technologists can be trained to perform high-quality esophagography and double contrast barium enema examinations without an unacceptably high radiation dose. PMID- 11044063 TI - Respiratory variation of the diameter of the pancreatic duct on sonography. AB - OBJECTIVE: The objective of this study was to assess the prevalence of a 1-mm or greater increase in the diameter of the pancreatic duct during deep inspiration in patients without pancreatic disease. MATERIALS AND METHODS: A retrospective review was performed of normal findings on pancreatic sonograms of 25 consecutive lean patients without pancreatic disease who were capable of taking deep breaths. The anteroposterior diameter of the pancreatic duct in the body of the gland was measured at end-expiration and end-inspiration. A significant change was defined as a 1-mm or greater difference between the end-inspiratory and end-expiratory diameters for at least two of three consecutive breaths. RESULTS: Seven patients (28%) had a significant increase in the diameter of the pancreatic duct at end inspiration. These included four patients (16%) in whom the diameter of the duct was less than or equal to 2.5 mm (i.e., normal) at end-inspiration and three patients (12%) in whom the diameter of the duct was greater than 2.5 mm at end inspiration. CONCLUSION: The diameter of the pancreatic duct can increase during deep inspiration in some adults without pancreatic disease. This finding should be borne in mind as a potential pitfall during pancreatic sonography. PMID- 11044062 TI - Dose reduction in gastrointestinal and genitourinary fluoroscopy: use of grid controlled pulsed fluoroscopy. AB - OBJECTIVE: We evaluated the diagnostic accuracy of a grid-controlled fluoroscopy unit compared with a conventional continuous fluoroscopy unit for a variety of abdominal and pelvic fluoroscopic examinations. SUBJECTS AND METHODS: Seventy patients (29 men and 41 women; age range, 24-78 years) were enrolled in one of seven abdominal and pelvic fluoroscopic examinations, including upper gastrointestinal series (n = 20), barium enema (n = 10), voiding cystourethrogram (n = 10), percutaneous abdominal catheter tube injection (n = 10), hysterosalpingogram (n = 10), and percutaneous needle insertion and catheter placement (nephrostomy, percutaneous biliary drainage) (n = 10). Each patient underwent at least 10 sec of continuous fluoroscopy that was randomly and blindly compared with 10-sec periods of pulsed fluoroscopy at 15, 7.5, and 3.75 frames per second. A radiologist outside the examination room, unaware of the frame rate per second, evaluated the procedure in real time on a television monitor. The radiologist assessed image quality and diagnostic acceptability using a scoring system. Statistical analysis was performed using the paired Student's t test. RESULTS: For all procedures at all frame rates, we found no statistically significant superiority of one frame rate over another. For most procedures, the slower frame rates were considered equivalent to continuous fluoroscopy when the images were assessed for image quality and diagnostic confidence. CONCLUSION: Our findings suggest that most abdominal and pelvic fluoroscopic procedures can be performed at substantially lower frame rates than those used for continuous fluoroscopy; adopting this procedure may lead to substantial dose savings for the patient and the fluoroscopy operator. PMID- 11044064 TI - MR imaging and MR angiography of pulmonary artery stenosis with tubular hypoplasia in pregnancy. PMID- 11044065 TI - Left-sided cisterna chyli. PMID- 11044066 TI - Anomalous unilateral single pulmonary vein: multidetector CT findings. PMID- 11044067 TI - Large villous adenoma in rectum mimicking cerebral hemispheres. PMID- 11044068 TI - MR imaging and endorectal sonographic appearance of a cyst of Skene's ducts. PMID- 11044069 TI - Selective angiography of cerebral aneurysm using gadodiamide in polycystic kidney disease with renal insufficiency. PMID- 11044070 TI - Reduced susceptibility of human immunodeficiency virus type 1 (HIV-1) from patients with primary HIV infection to nonnucleoside reverse transcriptase inhibitors is associated with variation at novel amino acid sites. AB - Recently, significant numbers of individuals with primary human immunodeficiency virus (HIV) infection have been found to harbor viral strains with reduced susceptibility to antiretroviral drugs. In one study, HIV from 16% of such antiretroviral-naive individuals was shown to have a susceptibility to nonnucleoside reverse transcriptase (RT) inhibitors (NNRTIs) between 2.5- and 10 fold lower than that of a wild-type control. Mutations in the RT domain that had previously been associated with antiretroviral resistance were not shared by these strains. We have analyzed by logistic regression 46 variable amino acid sites in RT for their effect on susceptibility and have identified two novel sites influencing susceptibility to NNRTIs: amino acids 135 and 283 in RT. Eight different combinations of amino acids at these sites were observed among these patients. These combinations showed a 14-fold range in mean susceptibility to both nevirapine and delavirdine. In vitro mutagenesis of the control strain combined with a phenotypic assay confirmed the significance of amino acid variation at these sites for susceptibility to NNRTIs. PMID- 11044072 TI - Efficacy and safety studies of a recombinant chimeric respiratory syncytial virus FG glycoprotein vaccine in cotton rats. AB - Several formulations of a recombinant chimeric respiratory syncytial virus (RSV) vaccine consisting of the extramembrane domains of the F and G glycoproteins (FG) were tested in cotton rats to evaluate efficacy and safety. The FG vaccine was highly immunogenic, providing nearly complete resistance to pulmonary infection at doses as low as 25 ng in spite of inducing relatively low levels of serum neutralizing antibody at low vaccine doses. Upon RSV challenge animals primed with FG vaccine showed quite mild alveolitis and interstitial pneumonitis, which were eliminated by the addition of monophosphoryl lipid A to the formulation. PMID- 11044071 TI - Dependence of adenovirus infectivity on length of the fiber shaft domain. AB - One of the objectives in adenovirus (Ad) vector development is to target gene delivery to specific cell types. Major attention has been given to modification of the Ad fiber knob, which is thought to determine virus tropism. However, among the human Ad serotypes with different tissue tropisms, not only the knob but also the length of the fiber shaft domain varies significantly. In this study we attempted to delineate the role of fiber length in coxsackievirus-adenovirus receptor (CAR)- and non-CAR-mediated infection. A series of Ad serotype 5 (Ad5) capsid-based vectors containing long or short fibers with knob domains derived from Ad5, Ad9, or Ad35 was constructed and tested in adsorption, internalization, and transduction studies. For Ad5 or Ad9 knob-possessing vectors, a long-shafted fiber was critical for efficient adsorption/internalization and transduction of CAR/alphav integrin-expressing cells. Ad5 capids containing short CAR-recognizing fibers were affected in cell adsorption and infection. In contrast, for the chimeric vectors possessing Ad35 knobs, which enter cells by a CAR/alphav integrin-independent pathway, fiber shaft length had no significant influence on binding or infectibility on tested cells. The weak attachment of short-shafted Ad5 or Ad9 knob-possessing vectors seems to be causally associated with a charge dependent repulsion between Ad5 capsid and acidic cell surface proteins. The differences between short- and long-shafted vectors in attachment or infection were abrogated by preincubation of cells with polycations. This study demonstrates that the fiber-CAR interaction is not the sole determinant for tropism of Ad vectors containing chimeric fibers. CAR- and alphav integrin mediated infections are influenced by other factors, including the length of the fiber shaft. PMID- 11044073 TI - Selection of optimal polypurine tract region sequences during Moloney murine leukemia virus replication. AB - Retrovirus plus-strand synthesis is primed by a cleavage remnant of the polypurine tract (PPT) region of viral RNA. In this study, we tested replication properties for Moloney murine leukemia viruses with targeted mutations in the PPT and in conserved sequences upstream, as well as for pools of mutants with randomized sequences in these regions. The importance of maintaining some purine residues within the PPT was indicated both by examining the evolution of random PPT pools and from the replication properties of targeted mutants. Although many different PPT sequences could support efficient replication and one mutant that contained two differences in the core PPT was found to replicate as well as the wild type, some sequences in the core PPT clearly conferred advantages over others. Contributions of sequences upstream of the core PPT were examined with deletion mutants. A conserved T-stretch within the upstream sequence was examined in detail and found to be unimportant to helper functions. Evolution of virus pools containing randomized T-stretch sequences demonstrated marked preference for the wild-type sequence in six of its eight positions. These findings demonstrate that maintenance of the T-rich element is more important to viral replication than is maintenance of the core PPT. PMID- 11044074 TI - Persistent virus infection despite chronic cytotoxic T-lymphocyte activation in gamma interferon-deficient mice infected with lymphocytic choriomeningitis virus. AB - The role of gamma interferon (IFN-gamma) in the permanent control of infection with a noncytopathic virus was studied by comparing immune responses in wild-type and IFN-gamma-deficient (IFN-gamma -/-) mice infected with a slowly invasive strain of lymphocytic choriomeningitis virus (LCMV Armstrong). While wild-type mice rapidly cleared the infection, IFN-gamma -/- mice became chronically infected. Virus persistence in the latter mice did not reflect failure to generate cytotoxic T-lymphocyte (CTL) effectors, as an unimpaired primary CTL response was observed. Furthermore, while ex vivo CTL activity gradually declined in wild-type mice, long-standing cytolytic activity was demonstrated in IFN-gamma -/- mice. The prolonged effector phase in infected IFN-gamma -/- mice was associated with elevated numbers of CD8(+) T cells. Moreover, a higher proportion of these cells retained an activated phenotype and was actively cycling. However, despite the increased CD8(+) T-cell turnover, which might have resulted in depletion of the memory CTL precursor pool, no evidence for exhaustion was observed. In fact, at 3 months postinfection we detected higher numbers of LCMV specific CTL precursors in IFN-gamma -/- mice than in wild-type mice. These findings indicate that in the absence of IFN-gamma, CTLs cannot clear the infection and are kept permanently activated by the continuous presence of live virus, resulting in a delicate new balance between viral load and immunity. This interpretation of our findings is supported by mathematical modeling describing the effect of eliminating IFN-gamma-mediated antiviral activity on the dynamics between virus replication and CTL activity. PMID- 11044075 TI - Template nucleotide moieties required for de novo initiation of RNA synthesis by a recombinant viral RNA-dependent RNA polymerase. AB - The recombinant RNA-dependent RNA polymerase of the bovine viral diarrhea virus specifically requires a cytidylate at the 3' end for the de novo initiation of RNA synthesis (C. C. Kao, A. M. Del Vecchio, and W. Zhong, Virology 253:1-7, 1999). Using RNAs containing nucleotide analogs, we found that the N3 and C4 amino group at the initiation cytidine were required for RNA synthesis. However, the ribose C2'-hydroxyl of the initiating cytidylate can accept several modifications and retain the ability to direct synthesis. The only unacceptable modification is a protonated C2'-amino group. Quite strikingly, the recognition of the functional groups for the initiation cytidylate and other template nucleotides are different. For example, a C5-methyl group in cytidine can direct RNA synthesis at all template positions except at the initiation cytidylate and C2'-amino modifications are tolerated better after the +11 position. When a 4 thiouracil (4sU) base analog that allows only imperfect base pairing with the nascent RNA is placed at different positions in the template, the efficiency of synthesis is correlated with the calculated stability of the template-nascent RNA duplex adjacent to the position of the 4sU. These results define the requirements for the specific interactions required for the initiation of RNA synthesis and will be compared to the mechanisms of initiation by other RNA-dependent and DNA dependent RNA polymerases. PMID- 11044076 TI - Recognition of the core RNA promoter for minus-strand RNA synthesis by the replicases of Brome mosaic virus and Cucumber mosaic virus. AB - Replication of viral RNA genomes requires the specific interaction between the replicase and the RNA template. Members of the Bromovirus and Cucumovirus genera have a tRNA-like structure at the 3' end of their genomic RNAs that interacts with the replicase and is required for minus-strand synthesis. In Brome mosaic virus (BMV), a stem-loop structure named C (SLC) is present within the tRNA-like region and is required for replicase binding and initiation of RNA synthesis in vitro. We have prepared an enriched replicase fraction from tobacco plants infected with the Fny isolate of Cucumber mosaic virus (Fny-CMV) that will direct synthesis from exogenously added templates. Using this replicase, we demonstrate that the SLC-like structure in Fny-CMV plays a role similar to that of BMV SLC in interacting with the CMV replicase. While the majority of CMV isolates have SLC like elements similar to that of Fny-CMV, a second group displays sequence or structural features that are distinct but nonetheless recognized by Fny-CMV replicase for RNA synthesis. Both motifs have a 5'CA3' dinucleotide that is invariant in the CMV isolates examined, and mutational analysis indicates that these are critical for interaction with the replicase. In the context of the entire tRNA-like element, both CMV SLC-like motifs are recognized by the BMV replicase. However, neither motif can direct synthesis by the BMV replicase in the absence of other tRNA-like elements, indicating that other features of the CMV tRNA can induce promoter recognition by a heterologous replicase. PMID- 11044077 TI - Gene transfer using recombinant rabbit hemorrhagic disease virus capsids with genetically modified DNA encapsidation capacity by addition of packaging sequences from the L1 or L2 protein of human papillomavirus type 16. AB - The aim of this study was to produce gene transfer vectors consisting of plasmid DNA packaged into virus-like particles (VLPs) with different cell tropisms. For this purpose, we have fused the N-terminally truncated VP60 capsid protein of the rabbit hemorrhagic disease virus (RHDV) with sequences which are expected to be sufficient to confer DNA packaging and gene transfer properties to the chimeric VLPs. Each of the two putative DNA-binding sequences of major L1 and minor L2 capsid proteins of human papillomavirus type 16 (HPV-16) were fused at the N terminus of the truncated VP60 protein. The two recombinant chimeric proteins expressed in insect cells self-assembled into VLPs similar in size and appearance to authentic RHDV virions. The chimeric proteins had acquired the ability to bind DNA. The two chimeric VLPs were therefore able to package plasmid DNA. However, only the chimeric VLPs containing the DNA packaging signal of the L1 protein were able efficiently to transfer genes into Cos-7 cells at a rate similar to that observed with papillomavirus L1 VLPs. It was possible to transfect only a very limited number of RK13 rabbit cells with the chimeric RHDV capsids containing the L2-binding sequence. The chimeric RHDV capsids containing the L1-binding sequence transfer genes into rabbit and hare cells at a higher rate than do HPV-16 L1 VLPs. However, no gene transfer was observed in human cell lines. The findings of this study demonstrate that the insertion of a DNA packaging sequence into a VLP which is not able to encapsidate DNA transforms this capsid into an artificial virus that could be used as a gene transfer vector. This possibility opens the way to designing new vectors with different cell tropisms by inserting such DNA packaging sequences into the major capsid proteins of other viruses. PMID- 11044078 TI - Generation of subgenomic RNA directed by a satellite RNA associated with bamboo mosaic potexvirus: analyses of potexvirus subgenomic RNA promoter. AB - Satellite RNA of bamboo mosaic potexvirus (satBaMV), a single-stranded positive sense RNA encoding a nonstructural protein of 20 kDa (P20), depends on bamboo mosaic potexvirus (BaMV) for replication and encapsidation. A full-length cDNA clone of satBaMV was used to examine the sequences required for the synthesis of potexvirus subgenomic RNAs (sgRNAs). Subgenomic promoter-like sequences (SGPs), 107 nucleotides (nt) upstream from the capsid protein (CP) gene of BaMV-V, were inserted upstream of the start codon of the P20 gene of satBaMV. Insertion of SGPs gave rise to the synthesis of sgRNA of satBaMV in protoplasts of Nicotiana benthamiana and leaves of Chenopodium quinoa when coinoculated with BaMV-V genomic RNA. Moreover, both the satBaMV cassette and its sgRNA were encapsidated. From analysis of the SGPs by deletion mutation, we concluded that an SGP contains one core promoterlike sequence (nt -30 through +16), two upstream enhancers (nt 59 through -31 and -91 through -60), and one downstream enhancer (nt +17 through +52), when the transcription initiation site is taken as +1. Site-directed mutagenesis and compensatory mutation to disrupt and restore potential base pairing in the core promoter-like sequence suggest that the stem-loop structure is important for the function of SGP in vivo. Likewise, the insertion of a putative SGP of the BaMV open reading frame 2 gene or a heterologous SGP of potato virus X resulted in generation of an sgRNA. The satBaMV cassette should be a useful tool to gain insight into sequences required for the synthesis of potexvirus sgRNAs. PMID- 11044079 TI - Role of murine leukemia virus reverse transcriptase deoxyribonucleoside triphosphate-binding site in retroviral replication and in vivo fidelity. AB - Retroviral populations exhibit a high evolutionary potential, giving rise to extensive genetic variation. Error-prone DNA synthesis catalyzed by reverse transcriptase (RT) generates variation in retroviral populations. Structural features within RTs are likely to contribute to the high rate of errors that occur during reverse transcription. We sought to determine whether amino acids within murine leukemia virus (MLV) RT that contact the deoxyribonucleoside triphosphate (dNTP) substrate are important for in vivo fidelity of reverse transcription. We utilized the previously described ANGIE P encapsidating cell line, which expresses the amphotropic MLV envelope and a retroviral vector (pGA 1). pGA-1 expresses the bacterial beta-galactosidase gene (lacZ), which serves as a reporter of mutations. Extensive mutagenesis was performed on residues likely to interact with the dNTP substrate, and the effects of these mutations on the fidelity of reverse transcription were determined. As expected, most substitution mutations of amino acids that directly interact with the dNTP substrate significantly reduced viral titers (>10,000-fold), indicating that these residues played a critical role in catalysis and viral replication. However, the D153A and A154S substitutions, which are predicted to affect the interactions with the triphosphate, resulted in statistically significant increases in the mutation rate. In addition, the conservative substitution F155W, which may affect interactions with the base and the ribose, increased the mutation rate 2.8-fold. Substitutions of residues in the vicinity of the dNTP-binding site also resulted in statistically significant decreases in fidelity (1. 3- to 2.4-fold). These results suggest that mutations of residues that contact the substrate dNTP can affect viral replication as well as alter the fidelity of reverse transcription. PMID- 11044081 TI - Genetic and biochemical studies of poliovirus cis-acting replication element cre in relation to VPg uridylylation. AB - In addition to highly conserved stem-loop structures located in the 5'- and 3' nontranslated regions, genome replication of picornaviruses requires cis-acting RNA elements located in the coding region (termed cre) (K. L. McKnight and S. M. Lemon, J. Virol. 70:1941-1952, 1996; P. E. Lobert, N. Escriou, J. Ruelle, and T. Michiels, Proc. Natl. Acad. Sci. USA 96:11560-11565, 1999; I. Goodfellow, Y. Chaudhry, A. Richardson, J. Meredith, J. W. Almond, W. Barclay, and D. J. Evans, J. Virol. 74:4590-4600, 2000). cre elements appear to be essential for minus strand RNA synthesis by an as-yet-unknown mechanism. We have discovered that the cre element of poliovirus (mapping to the 2C coding region of poliovirus type 1; nucleotides 4444 to 4505 in 2C), which is homologous to the cre element of poliovirus type 3, is preferentially used as a template for the in vitro uridylylation of VPg catalyzed by 3D(pol) in a reaction that is greatly stimulated by 3CD(pro) (A. V. Paul, E. Rieder, D. W. Kim, J. H. van Boom, and E. Wimmer, J. Virol. 74:10359-10370, 2000). Here we report a direct correlation between mutations that eliminate, or severely reduce, the in vitro VPg uridylylation reaction and produce replication phenotypes in vivo. None of the genetic changes significantly influenced translation or polyprotein processing. A substitution mapping to the first A (A4472C) of a conserved AAACA sequence in the loop of PV-cre(2C) eliminated the ability of the cre RNA to serve as template for VPg uridylylation and abolished RNA infectivity. Mutagenesis of the second A (A4473C; AAACA) severely reduced the yield of VPgpUpU and RNA infectivity was restored only after reversion to the wild-type sequence. The effect of substitution of the third A (A4474G; AAACA) was less severe but reduced both VPg uridylylation and virus yield. Disruption of base pairing within the upper stem region of PV-cre(2C) also affected uridylylation of VPg. Virus derived from transcripts containing mutations in the stem was either viable or quasi infectious. PMID- 11044080 TI - Identification of an RNA hairpin in poliovirus RNA that serves as the primary template in the in vitro uridylylation of VPg. AB - The first step in the replication of the plus-stranded poliovirus RNA is the synthesis of a complementary minus strand. This process is initiated by the covalent attachment of UMP to the terminal protein VPg, yielding VPgpU and VPgpUpU. We have previously shown that these products can be made in vitro in a reaction that requires only synthetic VPg, UTP, poly(A), purified poliovirus RNA polymerase 3D(pol), and Mg(2+) (A. V. Paul, J. H. van Boom, D. Filippov, and E. Wimmer, Nature 393:280-284, 1998). Since such a poly(A)-dependent process cannot confer sufficient specificity to poliovirus RNA replication, we have developed a new assay to search for a viral RNA template in conjunction with viral or cellular factors that could provide this function. We have now discovered a small RNA hairpin in the coding region of protein 2C as the site in PV1(M) RNA that is used as the primary template for the in vitro uridylylation of VPg. This hairpin has recently been described in poliovirus RNA as being an essential structure for the initiation of minus strand RNA synthesis (I. Goodfellow, Y. Chaudhry, A. Richardson, J. Meredith, J. W. Almond, W. Barclay, and D. J. Evans, J. Virol. 74:4590-4600, 2000). The uridylylation reaction either with transcripts of cre(2C) RNA or with full-length PV1(M) RNA as the template is strongly stimulated by the addition of purified viral protein 3CD(pro). Deletion of the cre(2C) RNA sequences from minigenomes eliminates their ability to serve as template in the reaction. A similar signal in the coding region of VP1 in HRV14 RNA (K. L. McKnight and S. M. Lemon, RNA 4:1569-1584, 1998) and the poliovirus cre(2C) can be functionally exchanged in the assay. The mechanism by which the VPgpUpU precursor, made specifically on the cre(2C) template, might be transferred to the site where it serves as primer for poliovirus RNA synthesis, remains to be determined. PMID- 11044082 TI - Adeno-associated virus (AAV) Rep protein enhances the generation of a recombinant mini-adenovirus (Ad) utilizing an Ad/AAV hybrid virus. AB - Mini-adenoviruses (mAd) deleted of all viral coding regions represent an emerging approach for transgene expression. We have exploited the unique features of the adeno-associated virus (AAV) terminal repeats within the context of an adenovirus adeno-associated hybrid virus (Ad/AAV) as a strategy for rapid and efficient generation of mAd. Excision and generation of mAd from the parental Ad/AAV hybrid vector was achieved in 293 cells through recombination but without selection for mAd production. Analysis of mAd isolated from 293 cells indicated that mAd DNA exists as monomer and dimer forms within the recombinant viral capsid. Formation of recombinant mAd was significantly increased using an AAV Rep78- or Rep68 expressing cell line through Rep-mediated excision utilizing the AAV terminal repeat sequences present in the Ad/AAV hybrid virus genome. The mAd viruses were infectious and able to transfer functional gene to A549 and HeLa cells. This approach is rapid and efficient, thereby providing a simplified methodology for generating mAd with functional transducing capabilities. PMID- 11044083 TI - The DNA of a plant retroviroid-like element is fused to different sites in the genome of a plant pararetrovirus and shows multiple forms with sequence deletions. AB - Carnation small viroid-like RNA (CarSV RNA) and its homologous DNA are the two forms of a unique plant retroviroid-like system. CarSV RNA is a 275-nucleotide noninfectious viroid-like RNA, present in certain carnation plants, which can adopt hammerhead structures in both polarity strands and self-cleave accordingly. CarSV DNA is organized as a series of head-to-tail multimers forming part of extrachromosomal elements in which CarSV DNA sequences are fused to sequences of carnation etched ring virus (CERV), a plant pararetrovirus. Analysis of more than 30 CarSV-CERV DNA junctions showed that distinct regions of the viral genome seem able to interact with CarSV DNA. All these junctions were short nucleotide stretches common to both CarSV and CERV DNAs. This suggests a polymerase-driven mechanism for their origin involving an enzyme with low processivity, most likely the viral reverse transcriptase. This view was further supported by the observation that most of CarSV sequences forming part of the junctions correspond either to strong secondary structure motifs in the conformation proposed for CarSV RNA or to its self-cleavage sites, which may have facilitated polymerase jumping. Accompanying the most-abundant CarSV RNA, a series of CarSV RNAs with sequence deletions were previously characterized. Here we have identified some of their corresponding DNA forms, together with other CarSV DNA forms with deletions not found in any CarSV RNA species identified so far. Some of these CarSV DNA forms have also been detected fused to CERV sequences. The existence of these shortened CarSV DNA versions may provide a continuous input of their corresponding transcripts and explain the persistence of CarSV RNAs with defective hammerhead structures for which an RNA-RNA model of amplification seems unlikely. PMID- 11044084 TI - Toward a comprehensive phylogeny for mammalian and avian herpesviruses. AB - With the aim of deriving a definitive phylogenetic tree for as many mammalian and avian herpesvirus species as possible, alignments were made of amino acid sequences from eight conserved and ubiquitously present genes of herpesviruses, with 48 virus species each represented by at least one gene. Phylogenetic trees for both single-gene and concatenated alignments were evaluated thoroughly by maximum-likelihood methods, with each of the three herpesvirus subfamilies (the Alpha-, Beta-, and Gammaherpesvirinae) examined independently. Composite trees were constructed starting with the top-scoring tree based on the broadest set of genes and supplemented by addition of virus species from trees based on narrower gene sets, to give finally a 46-species tree; branching order for three regions within the tree remained unresolved. Sublineages of the Alpha- and Betaherpesvirinae showed extensive cospeciation with host lineages by criteria of congruence in branching patterns and consistency in extent of divergence. The Gammaherpesvirinae presented a more complex picture, with both higher and lower substitution rates in different sublineages. The final tree obtained represents the most detailed view to date of phylogenetic relationships in any family of large-genome viruses. PMID- 11044085 TI - Human monoclonal antibodies that inhibit binding of hepatitis C virus E2 protein to CD81 and recognize conserved conformational epitopes. AB - The intrinsic variability of hepatitis C virus (HCV) envelope proteins E1 and E2 complicates the identification of protective antibodies. In an attempt to identify antibodies to E2 proteins from divergent HCV isolates, we produced HCV E2 recombinant proteins from individuals infected with HCV genotypes 1a, 1b, 2a, and 2b. These proteins were then used to characterize 10 human monoclonal antibodies (HMAbs) produced from peripheral B cells isolated from an individual infected with HCV genotype 1b. Nine of the antibodies recognize conformational epitopes within HCV E2. Six HMAbs identify epitopes shared among HCV genotypes 1a, 1b, 2a, and 2b. Six, including five broadly reactive HMAbs, could inhibit binding of HCV E2 of genotypes 1a, 1b, 2a, and 2b to human CD81 when E2 and the antibody were simultaneously exposed to CD81. Surprisingly, all of the antibodies that inhibited the binding of E2 to CD81 retained the ability to recognize preformed CD81-E2 complexes generated with some of the same recombinant E2 proteins. Two antibodies that did not recognize preformed complexes of HCV 1a E2 and CD81 also inhibited binding of HCV 1a virions to CD81. Thus, HCV-infected individuals can produce antibodies that recognize conserved conformational epitopes and inhibit the binding of HCV to CD81. The inhibition is mediated via antibody binding to epitopes outside of the CD81 binding site in E2, possibly by preventing conformational changes in E2 that are required for CD81 binding. PMID- 11044087 TI - Hepatitis C virus internal ribosome entry site (IRES) stem loop IIId contains a phylogenetically conserved GGG triplet essential for translation and IRES folding. AB - The hepatitis C virus (HCV) internal ribosome entry site (IRES) is a highly structured RNA element that directs cap-independent translation of the viral polyprotein. Morpholino antisense oligonucleotides directed towards stem loop IIId drastically reduced HCV IRES activity. Mutagenesis studies of this region showed that the GGG triplet (nucleotides 266 through 268) of the hexanucleotide apical loop of stem loop IIId is essential for IRES activity both in vitro and in vivo. Sequence comparison showed that apical loop nucleotides (UUGGGU) were absolutely conserved across HCV genotypes and the GGG triplet was strongly conserved among related Flavivirus and Pestivirus nontranslated regions. Chimeric IRES elements with IIId derived from GB virus B (GBV-B) in the context of the HCV IRES possess translational activity. Mutations within the IIId stem loop that abolish IRES activity also affect the RNA structure in RNase T(1)-probing studies, demonstrating the importance of correct RNA folding to IRES function. PMID- 11044086 TI - Ras-GAP binding and phosphorylation by herpes simplex virus type 2 RR1 PK (ICP10) and activation of the Ras/MEK/MAPK mitogenic pathway are required for timely onset of virus growth. AB - We used a herpes simplex virus type 2 (HSV-2) mutant with a deletion in the RR1 (ICP10) PK domain (ICP10DeltaPK) and an MEK inhibitor (PD98059) to examine the role of ICP10 PK in virus growth. In HSV-2-infected cells, ICP10 PK binds and phosphorylates the GTPase activating protein Ras-GAP. In vitro binding and peptide competition assays indicated that Ras-GAP N-SH2 and PH domains, respectively, bind ICP10 at phosphothreonines 117 and 141 and a WD40-like motif at positions 160 to 173. Binding and phosphorylation did not occur in cells infected with ICP10DeltaPK. GTPase activity was significantly lower in HSV-2- than in ICP10DeltaPK-infected cells. Conversely, the levels of activated Ras and mitogen-activated protein kinase (MAPK), and the expression and stabilization of the transcription factor c-Fos were significantly increased in cells infected with HSV-2 or a revertant virus [HSV-2(R)] but not with ICP10DeltaPK. PD98059 inhibited MAPK activation and induction-stabilization of c-Fos. Expression from the ICP10 promoter was increased in cells infected with HSV-2 but not with ICP10DeltaPK, and increased expression was ablated by PD98059. ICP10 DNA formed a complex with nuclear extracts from HSV-2-infected cells which was supershifted by c-Fos antibody and was not seen with extracts from ICP10DeltaPK-infected cells. Complex formation was abrogated by PD98059. Onset of HSV-2 replication was significantly delayed by PD98059 (14 h versus 2 h in untreated cells), a delay similar to that seen for ICP10DeltaPK. The data indicate that Ras-GAP phosphorylation by ICP10 PK is involved in the activation of the Ras/MEK/MAPK mitogenic pathway and c-Fos induction and stabilization. This results in increased ICP10 expression and the timely onset of HSV-2 growth. PMID- 11044088 TI - Borna disease virus persistence causes inhibition of glutamate uptake by feline primary cortical astrocytes. AB - Borna disease virus (BDV), a nonsegmented, negative-stranded (NNS) RNA virus, causes central nervous system (CNS) disease in a broad range of vertebrate species, including felines. Both viral and host factors contribute to very diverse clinical and pathological manifestations associated with BDV infection. BDV persistence in the CNS can cause neurobehavioral and neurodevelopmental abnormalities in the absence of encephalitis. These BDV-induced CNS disturbances are associated with altered cytokine and neurotrophin expression, as well as cell damage that is very restricted to specific brain regions and neuronal subpopulations. BDV also targets astrocytes, resulting in the development of prominent astrocytosis. Astrocytes play essential roles in maintaining CNS homeostasis, and disruption of their normal activities can contribute to altered brain function. Therefore, we have examined the effect of BDV infection on the astrocyte's physiology. We present here evidence that BDV can establish a nonlytic chronic infection in primary cortical feline astrocytes that is associated with a severe impairment in the astrocytes' ability to uptake glutamate. In contrast, the astrocytes' ability to uptake glucose, as well as their protein synthesis, viability, and rate of proliferation, was not affected by BDV infection. These findings suggest that, in vivo, BDV could also affect an important astrocyte function required to prevent neuronal excitotoxicity. This, in turn, might contribute to the neuropathogenesis of BDV. PMID- 11044090 TI - The Epstein-Barr virus promoter initiating B-cell transformation is activated by RFX proteins and the B-cell-specific activator protein BSAP/Pax5. AB - Epstein-Barr virus (EBV)-induced B-cell growth transformation, a central feature of the virus' strategy for colonizing the human B-cell system, requires full virus latent gene expression and is initiated by transcription from the viral promoter Wp. Interestingly, when EBV accesses other cell types, this growth transforming program is not activated. The present work focuses on a region of Wp which in reporter assays confers B-cell-specific activity. Bandshift studies indicate that this region contains three factor binding sites, termed sites B, C, and D, in addition to a previously characterized CREB site. Here we show that site C binds members of the ubiquitously expressed RFX family of proteins, notably RFX1, RFX3, and the associated factor MIBP1, whereas sites B and D both bind the B-cell-specific activator protein BSAP/Pax5. In reporter assays with mutant Wp constructs, the loss of factor binding to any one of these sites severely impaired promoter activity in B cells, while the wild-type promoter could be activated in non-B cells by ectopic BSAP expression. We suggest that Wp regulation by BSAP helps to ensure the B-cell specificity of EBV's growth transforming function. PMID- 11044089 TI - Immunization of cats against feline immunodeficiency virus (FIV) infection by using minimalistic immunogenic defined gene expression vector vaccines expressing FIV gp140 alone or with feline interleukin-12 (IL-12), IL-16, or a CpG motif. AB - Four groups of cats, each containing four animals, were immunized at 0, 3, and 6 weeks with minimalistic immunogenic defined gene expression vector (MIDGE) vaccines containing the gene(s) for feline immunodeficiency virus (FIV) gp140, FIV gp140 and feline interleukin-12 (IL-12), FIV gp140 and feline IL-16, or FIV gp140 and a CpG motif. MIDGEs were coated onto gold beads and injected intradermally with a gene gun. A fifth group of four cats were immunized in an identical manner but with blank gold beads. All cats were challenge exposed to virulent FIV 4 weeks following the final immunization, and the course of infection was monitored. The two groups of cats immunized with the FIV gp140 gene alone or with blank gold particles became highly viremic and seroconverted as early as 4 weeks after infection. In contrast, three of four cats immunized with FIV gp140 in combination with feline IL-12 failed to become viremic or seropositive, as has been shown elsewhere (F. S. Boretti, C. M. Leutenegger, C. Mislin, et al., AIDS 14:1749-1757, 2000). Here we show the effect of IL-12 when used as an adjuvant on the viral RNA and DNA load and on the cytokine profile. In addition, the two groups of cats immunized either with gp140 and IL-16 or with gp140 and the CpG had greatly reduced viremia. Protection correlated weakly with cytotoxic T-lymphocyte activity and increased cytokine transcription of IL-12, gamma interferon, and IL-10 by peripheral blood mononuclear cells in the postchallenge period. This study extends the data on IL-12 and provides new results on CpG motifs and IL-16 used as adjuvants in the FIV cat model. PMID- 11044091 TI - Methylation of transcription factor binding sites in the Epstein-Barr virus latent cycle promoter Wp coincides with promoter down-regulation during virus induced B-cell transformation. AB - Two Epstein-Barr virus latent cycle promoters for nuclear antigen expression, Wp and Cp, are activated sequentially during virus-induced transformation of B cells to B lymphoblastoid cell lines (LCLs) in vitro. Previously published restriction enzyme studies have indicated hypomethylation of CpG dinucleotides in the Wp and Cp regions of the viral genome in established LCLs, whereas these same regions appeared to be hypermethylated in Burkitt's lymphoma cells, where Wp and Cp are inactive. Here, using the more sensitive technique of bisulfite genomic sequencing, we reexamined the situation in established LCLs with the typical pattern of dominant Cp usage; surprisingly, this showed substantial methylation in the 400-bp regulatory region upstream of the Wp start site. This was not an artifact of long-term in vitro passage, since, in cultures of recently infected B cells, we found progressive methylation of Wp (but not Cp) regulatory sequences occurring between 7 and 21 days postinfection, coincident with the period in which dominant nuclear antigen promoter usage switches from Wp to Cp. Furthermore, in the equivalent in vivo situation, i.e., in the circulating B cells of acute infectious mononucleosis patients undergoing primary EBV infection, we again frequently observed selective methylation of Wp but not Cp sequences. An effector role for methylation in Wp silencing was supported by methylation cassette assays of Wp reporter constructs and by bandshift assays, where the binding of two sets of transcription factors important for Wp activation in B cells, BSAP/Pax5 and CREB/ATF proteins, was shown to be blocked by methylation of their binding sites. PMID- 11044092 TI - Identification of a membrane targeting and degradation signal in the p42 protein of influenza C virus. AB - Two mRNA species are derived from the influenza C virus RNA segment six, (i) a colinear transcript containing a 374-amino-acid residue open reading frame (referred to herein as the seg 6 ORF) which is translated to yield the p42 protein, and (ii) a spliced mRNA which encodes the influenza C virus matrix (CM1) protein consisting of the first 242 amino acids of p42. The p42 protein undergoes proteolytic cleavage at a consensus signal peptidase cleavage site after residue 259, yielding the p31 and CM2 proteins. Translocation of p42 into the endoplasmic reticulum membrane occurs cotranslationally and requires the hydrophobic internal signal peptide (residues 239 to 259), as well as the predicted transmembrane domain of CM2 (residues 285 to 308). The p31 protein was found to undergo rapid degradation after cleavage from p42. Addition of the 26S proteasome inhibitor lactacystin to influenza C virus-infected or seg 6 ORF cDNA-transfected cells drastically reduced p31 degradation. Transfer of the 17-residue C-terminal region of p31 to heterologous proteins resulted in their rapid turnover. The hydrophobic nature, but not the specific amino acid sequence of the 17-amino-acid C terminus of p31 appears to act as the signal for targeting the protein to membranes and for degradation. PMID- 11044093 TI - Pathogenic simian/human immunodeficiency virus SHIV(KU) inoculated into immunized macaques caused infection, but virus burdens progressively declined with time. AB - Using the simian immunodeficiency virus/human immunodeficiency virus (SHIV) macaque model of AIDS, we had shown in a previous report that a live, nonpathogenic strain of SHIV, further attenuated by deletion of the vpu gene and inoculated orally into adult macaques, had effectively prevented AIDS following vaginal inoculation with pathogenic SHIV(KU). Examination of lymph nodes from the animals at 18 weeks postchallenge had shown that all six animals were persistently infected with challenge virus. We report here on a 2-year follow-up study on the nature of the persistent infections in these animals. DNA of the vaccine virus was present in the lymph nodes at all time points tested, as far as 135 weeks postchallenge. In contrast, the DNA of SHIV(KU) became undetectable in one animal by week 55 and in three others by week 63. These four macaques have remained negative for SHIV(KU) DNA as far as the last time point examined at week 135. Quantification of the total viral DNA concentration in lymph nodes during the observation period showed a steady decline. All animals developed neutralizing antibody and cytotoxic-T-lymphocyte responses to SHIV(KU) that persisted throughout the observation period. Vaccine-like viruses were isolated from two animals, and a SHIV(KU)-like virus was isolated from one of the two macaques that remained positive for SHIV(KU) DNA. There was no evidence of recombination between the vaccine and the challenge viruses. Thus, immunization with the live vaccine not only prevented disease but also contributed to the steady decline in the virus burdens in the animals. PMID- 11044094 TI - Unprecedented degree of human immunodeficiency virus type 1 (HIV-1) group M genetic diversity in the Democratic Republic of Congo suggests that the HIV-1 pandemic originated in Central Africa. AB - The purpose of this study was to document the genetic diversity of human immunodeficiency virus type 1 (HIV-1) in the Democratic Republic of Congo (DRC; formerly Zaire). A total of 247 HIV-1-positive samples, collected during an epidemiologic survey conducted in 1997 in three regions (Kinshasa [the capital], Bwamanda [in the north], and Mbuyi-Maya [in the south]), were genetically characterized in the env V3-V5 region. All known subtypes were found to cocirculate, and for 6% of the samples the subtype could not be identified. Subtype A is predominant, with prevalences decreasing from north to south (69% in the north, 53% in the capital city, and 46% in the south). Subtype C, D, G, and H prevalences range from 7 to 9%, whereas subtype F, J, K, and CRF01-AE strains represent 2 to 4% of the samples; only one subtype B strain was identified. The highest prevalence (25%) of subtype C was in the south, and CRF01-AE was seen mainly in the north. The high intersubtype variability among the V3-V5 sequences is the most probable reason for the low (45%) efficiency of subtype A-specific PCR and HMA (heteroduplex mobility assay). Eighteen (29%) of 62 samples had discordant subtype designations between env and gag. Sequence analysis of the entire envelope from 13 samples confirmed the high degree of diversity and complexity of HIV-1 strains in the DRC; 9 had a complex recombinant structure in gp160, involving fragments of known and unknown subtypes. Interestingly, the unknown fragments from the different strains did not cluster together. Overall, the high number of HIV-1 subtypes cocirculating, the high intrasubtype diversity, and the high numbers of possible recombinant viruses as well as different unclassified strains are all in agreement with an old and mature epidemic in the DRC, suggesting that this region is the epicenter of HIV-1 group M. PMID- 11044095 TI - Glycosaminoglycan sulfation requirements for respiratory syncytial virus infection. AB - Glycosaminoglycans (GAGs) on the surface of cultured cells are important in the first step of efficient respiratory syncytial virus (RSV) infection. We evaluated the importance of sulfation, the major biosynthetic modification of GAGs, using an improved recombinant green fluorescent protein-expressing RSV (rgRSV) to assay infection. Pretreatment of HEp-2 cells with 50 mM sodium chlorate, a selective inhibitor of sulfation, for 48 h prior to inoculation reduced the efficiency of rgRSV infection to 40%. Infection of a CHO mutant cell line deficient in N sulfation was three times less efficient than infection of the parental CHO cell line, indicating that N-sulfation is important. In contrast, infection of a cell line deficient in 2-O-sulfation was as efficient as infection of the parental cell line, indicating that 2-O-sulfation is not required for RSV infection. Incubating RSV with the purified soluble heparin, the prototype GAG, before inoculation had previously been shown to neutralize its infectivity. Here we tested chemically modified heparin chains that lack their N-, C6-O-, or C2-O sulfate groups. Only heparin chains lacking the N-sulfate group lost the ability to neutralize infection, confirming that N-sulfation, but not C6-O- or C2-O sulfation, is important for RSV infection. Analysis of heparin fragments identified the 10-saccharide chain as the minimum size that can neutralize RSV infectivity. Taken together, these results show that, while sulfate modification is important for the ability of GAGs to mediate RSV infection, only certain sulfate groups are required. This specificity indicates that the role of cell surface GAGs in RSV infection is not based on a simple charge interaction between the virus and sulfate groups but instead involves a specific GAG structural configuration that includes N-sulfate and a minimum of 10 saccharide subunits. These elements, in addition to iduronic acid demonstrated previously (L. K. Hallak, P. L. Collins, W. Knudson, and M. E. Peeples, Virology 271:264-275, 2000), partially define cell surface molecules important for RSV infection of cultured cells. PMID- 11044096 TI - Effective induction of simian immunodeficiency virus-specific systemic and mucosal immune responses in primates by vaccination with proviral DNA producing intact but noninfectious virions. AB - We report a pilot evaluation of a DNA vaccine producing genetically inactivated simian immunodeficiency virus (SIV) particles in primates, with a focus on eliciting mucosal immunity. Our results demonstrate that DNA vaccines can be used to stimulate strong virus-specific mucosal immune responses in primates. The levels of immunoglobulin A (IgA) detected in rectal secretions of macaques that received the DNA vaccine intradermally and at the rectal mucosa were the most striking of all measured immune responses and were higher than usually achieved through natural infection. However, cytotoxic T lymphocyte responses were generally low and sporadically present in different animals. Upon rectal challenge with cloned SIVmac239, resistance to infection was observed, but some animals with high SIV-specific IgA levels in rectal secretions became infected. Our results suggest that high levels of IgA alone are not sufficient to prevent the establishment of chronic infection, although mucosal IgA responses may have a role in reducing the infectivity of the initial viral inoculum. PMID- 11044097 TI - High-mobility-group protein I can modulate binding of transcription factors to the U5 region of the human immunodeficiency virus type 1 proviral promoter. AB - HMG I/Y appears to be a multifunctional protein that relies on in its ability to interact with DNA in a structure-specific manner and with DNA, binding transcriptional activators via distinct protein-protein interaction surfaces. To investigate the hypothesis that HMG I/Y may have a role in human immunodeficiency virus type 1 (HIV-1) expression, we have analyzed whether HMG I/Y interacts with the 5' long terminal repeat and whether this interaction can modulate transcription factor binding. Using purified recombinant HMG I, we have identified several high-affinity binding sites which overlap important transcription factor binding sites. One of these HMG I binding sites coincides with an important binding site for AP-1 located downstream of the transcriptional start site, in the 5' untranslated region at the boundary of a positioned nucleosome. HMG I binding to this composite site inhibits the binding of recombinant AP-1. Consistent with this observation, using nuclear extracts prepared from Jurkat T cells, we show that HMG I (but not HMG Y) is strongly induced upon phorbol myristate acetate stimulation and this induced HMG I appears to both selectively inhibit the binding of basal DNA-binding proteins and enhance the binding of an inducible AP-1 transcription factor to this AP-1 binding site. We also report the novel finding that a component present in this inducible AP-1 complex is ATF-3. Taken together, these results argue that HMG I may play a fundamental role in HIV-1 expression by determining the nature of transcription factor-promoter interactions. PMID- 11044098 TI - Intracellular localization of vaccinia virus extracellular enveloped virus envelope proteins individually expressed using a Semliki Forest virus replicon. AB - The extracellular enveloped virus (EEV) form of vaccinia virus is bound by an envelope which is acquired by wrapping of intracellular virus particles with cytoplasmic vesicles containing trans-Golgi network markers. Six virus-encoded proteins have been reported as components of the EEV envelope. Of these, four proteins (A33R, A34R, A56R, and B5R) are glycoproteins, one (A36R) is a nonglycosylated transmembrane protein, and one (F13L) is a palmitylated peripheral membrane protein. During infection, these proteins localize to the Golgi complex, where they are incorporated into infectious virus that is then transported and released into the extracellular medium. We have investigated the fates of these proteins after expressing them individually in the absence of vaccinia infection, using a Semliki Forest virus expression system. Significant amounts of proteins A33R and A56R efficiently reached the cell surface, suggesting that they do not contain retention signals for intracellular compartments. In contrast, proteins A34R and F13L were retained intracellularly but showed distributions different from that of the normal infection. Protein A36R was partially retained intracellularly, decorating both the Golgi complex and structures associated with actin fibers. A36R was also transported to the plasma membrane, where it accumulated at the tips of cell projections. Protein B5R was efficiently targeted to the Golgi region. A green fluorescent protein fusion with the last 42 C-terminal amino acids of B5R was sufficient to target the chimeric protein to the Golgi region. However, B5R-deficient vaccinia virus showed a normal localization pattern for other EEV envelope proteins. These results point to the transmembrane or cytosolic domain of B5R protein as one, but not the only, determinant of the retention of EEV proteins in the wrapping compartment. PMID- 11044099 TI - Tat protein of human immunodeficiency virus type 1 induces interleukin-10 in human peripheral blood monocytes: implication of protein kinase C-dependent pathway. AB - The clinical manifestations observed in human immunodeficiency virus type 1 (HIV 1)-infected patients are primarily due to the capacity of the virus and its components to inactivate the immune system. HIV-1 Tat protein could participate in this immune system disorder. This protein is secreted by infected cells of HIV infected patients and is free in the plasma, where it can interact and be taken up by both infected and noninfected cells. In asymptomatic patients infected by HIV-1, production of interleukin-10 (IL-10), a highly immunosuppressive cytokine, is associated with disease progression to AIDS. In the present work, we tested the capacity of Tat to induce IL-10 production by peripheral blood monocytes of healthy donors. The results show that Tat causes the production of IL-10 in a dose- and stimulation time-dependent manner. Investigations of the mechanisms involved in signal transduction show that (i) the calcium pathway is not or only slightly involved in Tat-induced IL-10 production, (ii) the protein kinase C pathway plays an essential role, and (iii) monocyte stimulation by Tat results in the intranuclear translocation of transcription factor NF-kappaB and in the induction of phosphorylation of the mitogen-activated protein kinases ERK1 and ERK2; activation of these two potential substrates of protein kinase C is required for the production of IL-10. Finally, our results suggest that the effect of Tat is exerted at the membrane level and that the active domain is located within N-terminal residues 1 to 45. This production of IL-10 induced by Tat could participate in the progression of HIV infection to AIDS. PMID- 11044100 TI - In vivo selection of protease cleavage sites by using chimeric Sindbis virus libraries. AB - Identifying protease cleavage sites contributes to our understanding of their specificity and biochemical properties and can help in designing specific inhibitors. One route to this end is the generation and screening of random libraries of cleavage sites. Both synthetic and phage-displayed libraries have been extensively used in vitro. We describe a novel system based on recombinant Sindbis virus which can be used to identify cleavage sites in vivo, thus eliminating the need for a purified enzyme and overcoming the problem of choosing the correct in vitro conditions. As a model we used the serine protease of the hepatitis C virus (HCV). We engineered the gene coding for this enzyme and two specific cleavage sites in the Sindbis virus structural gene and constructed libraries of viral genomes with a random sequence at either of the cleavage sites. The system was designed so that only viral genomes coding for sequences cleaved by the protease would produce viable viruses. With this system we selected viruses containing sequences mirroring those of the natural HCV protease substrates which were cleaved with comparable efficiencies. PMID- 11044101 TI - The leader RNA of coronavirus mouse hepatitis virus contains an enhancer-like element for subgenomic mRNA transcription. AB - While the 5' cis-acting sequence of mouse hepatitis virus (MHV) for genomic RNA replication has been determined in several defective interfering (DI) RNA systems, it remains elusive for subgenomic RNA transcription. Previous studies have shown that the leader RNA in the DI genome significantly enhances the efficiency of DI subgenomic mRNA transcription, indicating that the leader RNA is a cis-acting sequence for mRNA transcription. To further characterize the cis acting sequence, we made a series of deletion mutants, all but one of which have an additional deletion of the cis-acting signal for replication in the 5' untranslated region. This deletion effectively eliminated the replication of the DI-chloramphenicol acetyltransferase (CAT)-reporter, as demonstrated by the sensitive reverse transcription (RT)-PCR. The ability of these replication-minus mutants to transcribe subgenomic mRNAs was then assessed using the DI RNA-CAT reporter system. Results from both CAT activity and mRNA transcripts detected by RT-PCR showed that a 5'-proximal sequence of 35 nucleotides (nt) at nt 25 to 59 is a cis-acting sequence required for subgenomic RNA transcription, while the consensus repeat sequence of the leader RNA does not have such effect. Analyses of the secondary structure indicate that this 35-nt sequence forms two stem-loops conserved among MHVs. Deletion of this sequence abrogated transcriptional activity and disrupted the predicted stem-loops and overall RNA secondary structure at the 5' untranslated region, suggesting that the secondary structure formed by this 35-nt sequence may facilitate the downstream consensus sequence accessible for the discontinuous RNA transcription. This may provide a mechanism by which the 5' cis-acting sequence regulates subgenomic RNA transcription. The 5'-most 24 nt are not essential for transcription, while the 9 nt immediately downstream of the leader enhances RNA transcription. The sequence between nt 86 and 135 had little effect on transcription. This study thus defines the cis acting transcription signal at the 5' end of the DI genome. PMID- 11044102 TI - Characterization of three nef-defective human immunodeficiency virus type 1 strains associated with long-term nonprogression. Australian Long-Term Nonprogressor Study Group. AB - Long-term survivors (LTS) of human immunodeficiency virus type 1 (HIV-1) infection provide an opportunity to investigate both viral and host factors that influence the rate of disease progression. We have identified three HIV-1 infected individuals in Australia who have been infected for over 11 years with viruses that contain deletions in the nef and nef-long terminal repeat (nef/LTR) overlap regions. These viruses differ from each other and from other nef defective strains of HIV-1 previously identified in Australia. One individual, LTS 3, is infected with a virus containing a nef gene with a deletion of 29 bp from the nef/LTR overlap region, resulting in a truncated Nef open reading frame. In addition to the Nef defect, only viruses containing truncated Vif open reading frames of 37 or 69 amino acids could be detected in peripheral blood mononuclear cells isolated from this patient. LTS 3 had a viral load of less than 20 copies of RNA/ml of plasma. The other two long-term survivors, LTS 9 and LTS 11, had loads of less than 200 copies of RNA/ml of plasma and are infected with viruses with larger deletions in both the nef alone and nef/LTR overlap regions. These viruses contain wild-type vif, vpu, and vpr accessory genes. All three strains of virus had envelope sequences characteristic of macrophagetropic viruses. These findings further indicate the reduced pathogenic potential of nef-defective viruses. PMID- 11044103 TI - Development of multigene and regulated lentivirus vectors. AB - Previously we described safe and efficient three-component human immunodeficiency virus type 1 (HIV-1)-based gene transfer systems for delivery of genes into nondividing cells (H. Mochizuki, J. P. Schwartz, K. Tanaka, R. O. Brady, and J. Reiser, J. Virol. 72:8873-8883, 1998). To apply such vectors in anti-HIV gene therapy strategies and to express multiple proteins in single target cells, we have engineered HIV-1 vectors for the concurrent expression of multiple transgenes. Single-gene vectors, bicistronic vectors, and multigene vectors expressing up to three exogenous genes under the control of two or three different transcriptional units, placed within the viral gag-pol coding region and/or the viral nef and env genes, were designed. The genes encoding the enhanced version of green fluorescent protein (EGFP), mouse heat-stable antigen (HSA), and bacterial neomycin phosphotransferase were used as models whose expression was detected by fluorescence-activated cell sorting, fluorescence microscopy, and G418 selection. Coexpression of these reporter genes in contact inhibited primary human skin fibroblasts (HSFs) persisted for at least 6 weeks in culture. Coexpression of the HSA and EGFP reporter genes was also achieved following cotransduction of target cells using two separate lentivirus vectors encoding HSA and EGFP, respectively. For the regulated expression of transgenes, tetracycline (Tet)-regulatable lentivirus vectors encoding the reverse Tet transactivator (rtTA) and EGFP controlled by a Tet-responsive element (TRE) were constructed. A binary HIV-1-based vector system consisting of a lentivirus encoding rtTA and a second lentivirus harboring a TRE driving the EGFP reporter gene was also designed. Doxycycline-modulated expression of the EGFP transgene was confirmed in transduced primary HSFs. These versatile vectors can potentially be used in a wide range of gene therapy applications. PMID- 11044104 TI - Strategy for systematic assembly of large RNA and DNA genomes: transmissible gastroenteritis virus model. AB - A systematic method was developed to assemble functional full-length genomes of large RNA and DNA viruses. Coronaviruses contain the largest single-stranded positive-polarity RNA genome in nature. The approximately 30-kb genome, coupled with regions of genomic instability, has hindered the development of a full length infectious cDNA construct. We have assembled a full-length infectious construct of transmissible gastroenteritis virus (TGEV), an important pathogen in swine. Using a novel approach, six adjoining cDNA subclones that span the entire TGEV genome were isolated. Each clone was engineered with unique flanking interconnecting junctions which determine a precise systematic assembly with only the adjacent cDNA subclones, resulting in an intact TGEV cDNA construct of approximately 28.5 kb in length. Transcripts derived from the full-length TGEV construct were infectious, and progeny virions were serially passaged in permissive host cells. Viral antigen production and subgenomic mRNA synthesis were evident during infection and throughout passage. Plaque-purified virus derived from the infectious construct replicated efficiently and displayed similar plaque morphology in permissive host cells. Host range phenotypes of the molecularly cloned and wild-type viruses were similar in cells of swine and feline origin. The recombinant viruses were sequenced across the unique interconnecting junctions, conclusively demonstrating the marker mutations and restriction sites that were engineered into the component clones. Full-length infectious constructs of TGEV will permit the precise genetic modification of the coronavirus genome. The method that we have designed to generate an infectious cDNA construct of TGEV could theoretically be used to precisely reconstruct microbial or eukaryotic genomes approaching several million base pairs in length. PMID- 11044105 TI - Utilization of the bovine papillomavirus type 1 late-stage-specific nucleotide 3605 3' splice site is modulated by a novel exonic bipartite regulator but not by an intronic purine-rich element. AB - Bovine papillomavirus type 1 (BPV-1) late gene expression is regulated at both transcriptional and posttranscriptional levels. Maturation of the capsid protein (L1) pre-mRNA requires a switch in 3' splice site utilization. This switch involves activation of the nucleotide (nt) 3605 3' splice site, which is utilized only in fully differentiated keratinocytes during late stages of the virus life cycle. Our previous studies of the mechanisms that regulate BPV-1 alternative splicing identified three cis-acting elements between these two splice sites. Two purine-rich exonic splicing enhancers, SE1 and SE2, are essential for preferential utilization of the nt 3225 3' splice site at early stages of the virus life cycle. Another cis-acting element, exonic splicing suppressor 1 (ESS1), represses use of the nt 3225 3' splice site. In the present study, we investigated the late-stage-specific nt 3605 3' splice site and showed that it has suboptimal features characterized by a nonconsensus branch point sequence and a weak polypyrimidine track with interspersed purines. In vitro and in vivo experiments showed that utilization of the nt 3605 3' splice site was not affected by SE2, which is intronically located with respect to the nt 3605 3' splice site. The intronic location and sequence composition of SE2 are similar to those of the adenovirus IIIa repressor element, which has been shown to inhibit use of a downstream 3' splice site. Further studies demonstrated that the nt 3605 3' splice site is controlled by a novel exonic bipartite element consisting of an AC-rich exonic splicing enhancer (SE4) and an exonic splicing suppressor (ESS2) with a UGGU motif. Functionally, this newly identified bipartite element resembles the bipartite element composed of SE1 and ESS1. SE4 also functions on a heterologous 3' splice site. In contrast, ESS2 functions as an exonic splicing suppressor only in a 3'-splice-site-specific and enhancer-specific manner. Our data indicate that BPV-1 splicing regulation is very complex and is likely to be controlled by multiple splicing factors during keratinocyte differentiation. PMID- 11044106 TI - Expression of the two major envelope proteins of equine arteritis virus as a heterodimer is necessary for induction of neutralizing antibodies in mice immunized with recombinant Venezuelan equine encephalitis virus replicon particles. AB - RNA replicon particles derived from a vaccine strain of Venezuelan equine encephalitis virus (VEE) were used as a vector for expression of the major envelope proteins (G(L) and M) of equine arteritis virus (EAV), both individually and in heterodimer form (G(L)/M). Open reading frame 5 (ORF5) encodes the G(L) protein, which expresses the known neutralizing determinants of EAV (U. B. R. Balasuriya, J. F. Patton, P. V. Rossitto, P. J. Timoney, W. H. McCollum, and N. J. MacLachlan, Virology 232:114-128, 1997). ORF5 and ORF6 (which encodes the M protein) of EAV were cloned into two different VEE replicon vectors that contained either one or two 26S subgenomic mRNA promoters. These replicon RNAs were packaged into VEE replicon particles by VEE capsid protein and glycoproteins supplied in trans in cells that were coelectroporated with replicon and helper RNAs. The immunogenicity of individual replicon particle preparations (pVR21 G(L), pVR21-M, and pVR100-G(L)/M) in BALB/c mice was determined. All mice developed antibodies against the recombinant proteins with which they were immunized, but only the mice inoculated with replicon particles expressing the G(L)/M heterodimer developed antibodies that neutralize EAV. The data further confirmed that authentic posttranslational modification and conformational maturation of the recombinant G(L) protein occur only in the presence of the M protein and that this interaction is necessary for induction of neutralizing antibodies. PMID- 11044107 TI - Site-specific integration of an adeno-associated virus vector plasmid mediated by regulated expression of rep based on Cre-loxP recombination. AB - Recombinant adeno-associated virus (AAV) type 2 has attracted attention because it appears to have the potential to serve as a vector for human gene therapy. An interesting feature of wild-type AAV is its site-specific integration into AAVS1, a defined locus on chromosome 19. This reaction requires the presence of two viral elements: inverted terminal repeats and Rep78/68. Accordingly, current AAV vectors lacking the rep gene lack the capacity for site-specific integration. In this report, we describe the use of Cre-loxP recombination in a novel system for the regulated, transient expression of Rep78, which is potentially cytotoxic when synthesized constitutively. We constructed a plasmid in which the p5 promoter was situated downstream of the rep coding sequence; in this configuration, rep expression is silent. However, Cre circularizes the rep expression unit, directly joining the p5 promoter to the 5' end of the rep78 coding sequence, resulting in expression of Rep78. Such structural and functional changes were confirmed by detailed molecular analysis. A key feature of this system is that Rep expression was terminated when the circular molecule was linearized and integrated into the chromosome. Using this regulated expression system, we attempted site-specific integration of AAV vector plasmids. A PCR-based assay and analysis of fluorescence in situ hybridization showed that the AAV vector sequence was integrated into chromosome 19. Sequence analysis also confirmed that transient expression of Rep78 was sufficient for site-specific integration at the AAVS1 locus, as is observed with integration of wild-type AAV. PMID- 11044108 TI - Canine adenovirus type 2 attachment and internalization: coxsackievirus adenovirus receptor, alternative receptors, and an RGD-independent pathway. AB - The best-characterized receptors for adenoviruses (Ads) are the coxsackievirus-Ad receptor (CAR) and integrins alpha(v)beta(5) and alpha(v)beta(3), which facilitate entry. The alpha(v) integrins recognize an Arg-Gly-Asp (RGD) motif found in some extracellular matrix proteins and in the penton base in most human Ads. Using a canine adenovirus type 2 (CAV-2) vector, we found that CHO cells that express CAR but not wild-type CHO cells are susceptible to CAV-2 transduction. Cells expressing alpha(M)beta(2) integrins or major histocompatibility complex class I (MHC-I) molecules but which do not express CAR were not transduced. Binding assays showed that CAV-2 attaches to a recombinant soluble form of CAR and that Ad type 5 (Ad5) fiber, penton base, and an anti-CAR antibody partially blocked attachment. Using fluorescently labeled CAV-2 particles, we found that in some cells nonpermissive for transduction, inhibition was at the point of internalization and not attachment. The transduction efficiency of CAV-2, which lacks an RGD motif, surprisingly mimicked that of Ad5 when tested in cells selectively expressing alpha(v)beta(5) and alpha(v)beta(3) integrins. Our results demonstrate that CAV-2 transduction is augmented by CAR and possibly by alpha(v)beta(5), though transduction can be CAR and alpha(v)beta(3/5) independent but is alpha(M)beta(2), MHC-I, and RGD independent, demonstrating a transduction mechanism which is distinct from that of Ad2/5. PMID- 11044110 TI - Life cycle of an endogenous retrovirus, ZAM, in Drosophila melanogaster. AB - ZAM is an env-containing member of the gypsy family of retrotransposons that represents a possible retrovirus of invertebrates. In this paper, we traced ZAM mobilization to get information about a potential path a retroelement may take to reach the germ line of its host. In situ hybridization on whole-mount tissues and immunocytochemistry analyses with antibodies raised against ZAM Gag and Env proteins have shown that all components necessary to assemble ZAM viral particles, i.e., ZAM full-length RNAs and Gag and Env polypeptides, are coexpressed in a small set of follicle cells surrounding the oocyte. By electron microscopy, we have shown that ZAM viral particles are indeed detected in this somatic lineage of cells, which they leave and enter the closely apposed oocyte. Our data provide evidence that the vesicular traffic and yolk granules in the process of vitellogenesis play an important role in ZAM transfer to the oocyte. Our data support the possibility that vitellogenin transfer to the oocyte may help a retroelement pass to the germ line with no need of its envelope product. PMID- 11044109 TI - Functional role of residues corresponding to helical domain II (amino acids 35 to 46) of human immunodeficiency virus type 1 Vpr. AB - Vpr, encoded by the human immunodeficiency virus type 1 genome, contains 96 amino acids and is a multifunctional protein with features which include cell cycle arrest at G(2), nuclear localization, participation in transport of the preintegration complex, cation channel activity, oligomerization, and interaction with cellular proteins, in addition to its incorporation into the virus particles. Recently, structural studies based on nuclear magnetic resonance and circular dichroism spectroscopy showed that Vpr contains a helix (HI)-turn-helix (HII) core at the amino terminus and an amphipathic helix (HIII) in the middle region. Though the importance of helical domains HI and HIII has been defined with respect to Vpr functions, the role of helical domain HII is not known. To address this issue, we constructed a series of mutants in which the HII domain was altered by deletion, insertion, and/or substitution mutagenesis. To enable the detection of Vpr, the sequence corresponding to the Flag epitope (DYKDDDDK) was added, in frame, to the Vpr coding sequences. Mutants, expressed through the in vitro transcription/translation system and in cells, showed an altered migration corresponding to deletions in Vpr. Substitution mutational analysis of residues in HII showed reduced stability for VprW38S-FL, VprL42G-FL, and VprH45W FL. An assay involving cotransfection of NLDeltaVpr proviral DNA and a Vpr expression plasmid was employed to analyze the virion incorporation property of Vpr. Mutant Vpr containing deletions and specific substitutions (VprW38S-FL, VprL39G-FL, VprL42G-FL, VprG43P-FL, and VprI46G-FL) exhibited a negative virion incorporation phenotype. Further, mutant Vpr-FL containing deletions also failed to associate with wild-type Vpr, indicating a possible defect in the oligomerization feature of Vpr. Subcellular localization studies indicated that mutants VprDelta35-50-H-FL, VprR36W-FL, VprL39G-FL, and VprI46G-FL exhibited both cytoplasmic and nuclear localization, unlike other mutants and control Vpr-FL. While wild-type Vpr registered cell cycle arrest at G(2), mutant Vpr showed an intermediary effect with the exception of VprDelta35-50 and VprDelta35-50-H. These results suggest that residues in the HII domain are essential for Vpr functions. PMID- 11044111 TI - Immunoreactivity of intact virions of human immunodeficiency virus type 1 (HIV-1) reveals the existence of fewer HIV-1 immunotypes than genotypes. AB - In order to protect against organisms that exhibit significant genetic variation, polyvalent vaccines are needed. Given the extreme variability of human immunodeficiency virus type 1 (HIV-1), it is probable that a polyvalent vaccine will also be needed for protection from this virus. However, to understand how to construct a polyvalent vaccine, serotypes or immunotypes of HIV must be identified. In the present study, we have examined the immunologic relatedness of intact, native HIV-1 primary isolates of group M, clades A to H, with human monoclonal antibodies (MAbs) directed at epitopes in the V3, C5, and gp41 cluster I regions of the envelope glycoproteins, since these regions are well exposed on the virion surface. Multivariate analysis of the binding data revealed three immunotypes of HIV-1 and five MAb groups useful for immunotyping of the viruses. The analysis revealed that there are fewer immunotypes than genotypes of HIV and that clustering of the isolates did not correlate with either genotypes, coreceptor usage (CCR5 and CXCR4), or geographic origin of the isolates. Further analysis revealed distinct MAb groups that bound preferentially to HIV-1 isolates belonging to particular immunotypes or that bound to all three immunotypes; this demonstrates that viral immunotypes identified by mathematical analysis are indeed defined by their immunologic characteristics. In summary, these results indicate (i) that HIV-1 immunotypes can be defined, (ii) that constellations of epitopes that are conserved among isolates belonging to each individual HIV-1 immunotype exist and that these distinguish each of the immunotypes, and (iii) that there are also epitopes that are routinely shared by all immunotypes. PMID- 11044112 TI - Epstein-Barr virus LMP2A transforms epithelial cells, inhibits cell differentiation, and activates Akt. AB - The Epstein-Barr virus LMP2A protein was expressed in a human keratinocyte cell line, HaCaT, and effects on epithelial cell growth were detected in organotypic raft cultures and in vivo in nude mice. Raft cultures derived from LMP2A expressing cells were hyperproliferative, and epithelial differentiation was inhibited. The LMP2A-expressing HaCaT cells were able to grow anchorage independently and formed colonies in soft agar. HaCaT cells expressing LMP2A were highly tumorigenic and formed aggressive tumors in nude mice. The LMP2A tumors were poorly differentiated and highly proliferative, in contrast to occasional tumors that arose from parental HaCaT cells and vector control cells, which grew slowly and remained highly differentiated. Animals injected with LMP2A-expressing cells developed frequent metastases, which predominantly involved lymphoid organs. Involucrin, a marker of epithelial differentiation, and E-cadherin, involved in the maintenance of intercellular contact, were downregulated in LMP2A tumors. Whereas activation of the mitogen-activated protein kinase pathway was not observed, phosphatidylinositol-3-kinase (PI3-kinase)-dependent activation of the serine-threonine kinase Akt was detected in LMP2A-expressing cells and LMP2A tumors. Inhibition of this pathway blocked growth in soft agar. These data indicate that LMP2A greatly affects cell growth and differentiation pathways in epithelial cells, in part through activation of the PI3-kinase-Akt pathway. PMID- 11044113 TI - Importance of membrane fusion mediated by human immunodeficiency virus envelope glycoproteins for lysis of primary CD4-positive T cells. AB - In established T-cell lines, the membrane-fusing capacity of the human immunodeficiency virus type 1 (HIV-1) envelope glycoproteins mediates cytopathic effects, both syncytium formation and single-cell lysis. Furthermore, changes in the HIV-1 envelope glycoproteins are responsible for the increased CD4(+) T-cell depleting ability observed in infected monkeys upon in vivo passage of simian human immunodeficiency virus (SHIV) chimeras. In this study, a panel of SHIV envelope glycoproteins and their mutant counterparts defective in membrane-fusing capacity were expressed in primary human CD4(+) T cells. Compared with controls, all of the functional HIV-1 envelope glycoproteins induced cell death in primary CD4(+) T-cell cultures, whereas the membrane fusion-defective mutants did not. Death occurred almost exclusively in envelope glycoprotein-expressing cells and not in bystander cells. Under standard culture conditions, most dying cells underwent lysis as single cells. When the cells were cultured at high density to promote syncytium formation, the envelope glycoproteins of the passaged, pathogenic SHIVs induced more syncytia than those of the respective parental SHIV. These results demonstrate that the HIV-1 envelope glycoproteins induce the death of primary CD4(+) T lymphocytes by membrane fusion-dependent processes. PMID- 11044114 TI - Viral vascular endothelial growth factor plays a critical role in orf virus infection. AB - Infection by the parapoxvirus orf virus causes proliferative skin lesions in which extensive capillary proliferation and dilation are prominent histological features. This infective phenotype may be linked to a unique virus-encoded factor, a distinctive new member of the vascular endothelial growth factor (VEGF) family of molecules. We constructed a recombinant orf virus in which the VEGF like gene was disrupted and show that inactivation of this gene resulted in the loss of three VEGF activities expressed by the parent virus: mitogenesis of vascular endothelial cells, induction of vascular permeability, and activation of VEGF receptor 2. We used the recombinant orf virus to assess the contribution of the viral VEGF to the vascular response seen during orf virus infection of skin. Our results demonstrate that the viral VEGF, while recognizing a unique profile of the known VEGF receptors (receptor 2 and neuropilin 1), is able to stimulate a striking proliferation of blood vessels in the dermis underlying the site of infection. Furthermore, the data demonstrate that the viral VEGF participates in promoting a distinctive pattern of epidermal proliferation. Loss of a functional viral VEGF resulted in lesions with markedly reduced clinical indications of infection. However, viral replication in the early stages of infection was not impaired, and only at later times did it appear that replication of the recombinant virus might be reduced. PMID- 11044115 TI - Genotypic, phenotypic, and modeling studies of a deletion in the beta3-beta4 region of the human immunodeficiency virus type 1 reverse transcriptase gene that is associated with resistance to nucleoside reverse transcriptase inhibitors. AB - Point mutations and inserts in the beta3-beta4 region of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) are associated with resistance to nucleoside analog inhibitors. This report describes HIV-1 strains from seven patients that were found to have a 3-bp deletion in the beta3-beta4 region of the RT gene. These patient strains also had a mean of 6.2 drug resistance-associated mutations in their RT genes (range, 3 to 10 mutations). The deletion was most frequently found in strains with the Q151M mutation. Nonnucleoside RT inhibitor mutations were found in six of seven strains. Culture-based drug sensitivity assays showed that deletion-containing isolates had reduced susceptibility to four to eight RT inhibitors. Site-directed mutagenesis experiments showed that the deletion alone conferred reduced susceptibility to nucleoside analogs. Changes in the three-dimensional models of the RT deletion mutants were consistently observed at the beta3-beta4 loop and at helices C and E in both the presence and the absence of dTTP. Loss of hydrogen bonds between the RT and dTTP were also observed in the RT deletion mutant. These results suggest that the deletion in the RT gene contributes to resistance to several nucleoside analogs through a complex interaction with other mutations in the RT gene. PMID- 11044116 TI - Evolution of bovine respiratory syncytial virus. AB - Until now, the analysis of the genetic diversity of bovine respiratory syncytial virus (BRSV) has been based on small numbers of field isolates. In this report, we determined the nucleotide and deduced amino acid sequences of regions of the nucleoprotein (N protein), fusion protein (F protein), and glycoprotein (G protein) of 54 European and North American isolates and compared them with the sequences of 33 isolates of BRSV obtained from the databases, together with those of 2 human respiratory syncytial viruses and 1 ovine respiratory syncytial virus. A clustering of BRSV sequences according to geographical origin was observed. We also set out to show that a continuous evolution of the sequences of the N, G, and F proteins of BRSV has been occurring in isolates since 1967 in countries where vaccination was widely used. The exertion of a strong positive selective pressure on the mucin-like region of the G protein and on particular sites of the N and F proteins is also demonstrated. Furthermore, mutations which are located in the conserved central hydrophobic part of the ectodomain of the G protein and which result in the loss of four Cys residues and in the suppression of two disulfide bridges and an alpha helix critical to the three-dimensional structure of the G protein have been detected in some recent French BRSV isolates. This conserved central region, which is immunodominant in BRSV G protein, thus has been modified in recent isolates. This work demonstrates that the evolution of BRSV should be taken into account in the rational development of future vaccines. PMID- 11044117 TI - Comparison between human cytomegalovirus pUL97 and murine cytomegalovirus (MCMV) pM97 expressed by MCMV and vaccinia virus: pM97 does not confer ganciclovir sensitivity. AB - The UL97 protein (pUL97) of human cytomegalovirus (HCMV) is a protein kinase that also phosphorylates ganciclovir (GCV), but its biological function is not yet clear. The M97 protein (pM97) of mouse cytomegalovirus (MCMV) is the homolog of pUL97. First, we studied the consequences of genetic replacement of M97 by UL97. Using the infectious bacterial plasmid clone of the full-length MCMV genome (M. Wagner, S. Jonjic, U. H. Koszinowski, and M. Messerle, J. Virol. 73:7056-7060, 1999), we replaced the M97 gene with the UL97 gene and constructed an MCMV M97 deletion mutant and a revertant virus. In addition, pUL97 and pM97 were expressed by recombinant vaccinia virus to compare both for known functions. Remarkably, pM97 proved not to be the reason for the GCV sensitivity of MCMV. When expressed by the recombinant MCMV, however, pUL97 was phosphorylated and endowed MCMV with the capacity to phosphorylate GCV, thereby rendering MCMV more susceptible to GCV. We found that deletion of pM97, although it is not essential for MCMV replication, severely affected virus growth. This growth deficit was only partially amended by pUL97 expression. When expressed by recombinant vaccinia viruses, both proteins were phosphorylated and supported phosphorylation of GCV, but pUL97 was about 10 times more effective than pM97. One hint of the functional differences between the proteins was provided by the finding that pUL97 accumulates in the nucleus, whereas pM97 is predominantly located in the cytoplasm of infected cells. In vivo testing revealed that the UL97-MCMV recombinant should allow evaluation of novel antiviral drugs targeted to the UL97 protein of HCMV in mice. PMID- 11044118 TI - Establishment of a rescue system for canine distemper virus. AB - Canine distemper virus (CDV) has been rescued from a full-length cDNA clone. Besides Measles virus (MV) and Rinderpest virus, a third morbillivirus is now available for genetic analysis using reverse genetics. A plasmid p(+)CDV was constructed by sequential cloning using the Onderstepoort vaccine strain large plaque-forming variant. The presence of a T7 promoter allowed transcription of full-length antigenomic RNA by a T7 RNA polymerase, which was provided by a host range mutant of vaccinia virus (MVA-T7). Plasmids expressing the nucleocapsid protein, the phosphoprotein, and the viral RNA-dependent RNA polymerase, also under control of a T7 promoter, have been generated. Infection of HeLa cells with MVA-T7 and subsequent transfection of p(+)CDV plus the helper plasmids led to syncytium formation and release of infectious recombinant (r) CDV. Comparison of the rescued virus with the parental virus revealed no major differences in the progression of infection or in the shape and size of syncytia. A genetic tag, consisting of two nucleotide changes within the coding region of the L protein, has been identified in the rCDV genome. Expression by rCDV of all the major viral structural proteins has been demonstrated by immunofluorescence. PMID- 11044119 TI - CD21-mediated entry and stable infection by Epstein-Barr virus in canine and rat cells. AB - We developed an adenovirus vector for transduction of the human CD21 gene (Adv CD21), the Epstein-Barr virus (EBV)-specific receptor on human B lymphocytes, to overcome the initial barrier of EBV infection in nonprimate mammalian cells. Inoculation of Adv-CD21 followed by exposure to recombinant EBV carrying a selectable marker resulted in the successful entry of EBV into three of seven nonprimate mammalian cell lines as evidenced by expression of EBV-determined nuclear antigen (EBNA). The EBV-susceptible cell lines included rat glioma derived 9L, rat mammary carcinoma-derived c-SST-2, and canine kidney-derived MDCK. Subsequent selection culture with G418 yielded drug-resistant cell clones. In these cell clones, EBV existed as an episomal form, as evidenced through the Gardella gel technique. Among the known EBV latency-associated gene products, EBV encoded small RNAs, EBNA1 and transcripts from the BamHI-A rightward reading frame (BARF0), and latent membrane protein 2A were expressed in all EBV-infected cell clones. The viral lytic events could be induced in these cell clones by simultaneous treatment with 12-O-tetradecanoylphorbol-13-acetate and n-butyric acid, but they were abortive, and infectious virus was not produced. These results indicate that once the initial barrier for attachment is overcome artificially, EBV can establish a stable infection in some nonprimate mammalian cells, and they raise the possibility that transgenic animals with the human CD21 gene could provide an animal model for EBV infection. PMID- 11044120 TI - Testing the hypothesis of a recombinant origin of human immunodeficiency virus type 1 subtype E. AB - The human immunodeficiency virus type 1 (HIV-1) epidemic in Southeast Asia has been largely due to the emergence of clade E (HIV-1E). It has been suggested that HIV-1E is derived from a recombinant lineage of subtype A (HIV-1A) and subtype E, with multiple breakpoints along the E genome. We obtained complete genome sequences of clade E viruses from Thailand (93TH057 and 93TH065) and from the Central African Republic (90CF11697 and 90CF4071), increasing the total number of HIV-1E complete genome sequences available to seven. Phylogenetic analysis of complete genomes showed that subtypes A and E are themselves monophyletic, although together they also form a larger monophyletic group. The apparent phylogenetic incongruence at different regions of the genome that was previously taken as evidence of recombination is shown to be not statistically significant. Furthermore, simulations indicate that bootscanning and pairwise distance results, previously used as evidence for recombination, can be misleading, particularly when there are differences in substitution or evolutionary rates across the genomes of different subtypes. Taken jointly, our analyses suggest that there is inadequate support for the hypothesis that subtype E variants are derived from a recombinant lineage. In contrast, many other HIV strains claimed to have a recombinant origin, including viruses for which only a single parental strain was employed for analysis, do indeed satisfy the statistical criteria we propose. Thus, while intersubtype recombinant HIV strains are indeed circulating, the criteria for assigning a recombinant origin to viral structures should include statistical testing of alternative hypotheses to avoid inappropriate assignments that would obscure the true evolutionary properties of these viruses. PMID- 11044121 TI - Variation in the nucleotide sequence of cottontail rabbit papillomavirus a and b subtypes affects wart regression and malignant transformation and level of viral replication in domestic rabbits. AB - We previously reported the partial characterization of two cottontail rabbit papillomavirus (CRPV) subtypes with strikingly divergent E6 and E7 oncoproteins. We report now the complete nucleotide sequences of these subtypes, referred to as CRPVa4 (7,868 nucleotides) and CRPVb (7,867 nucleotides). The CRPVa4 and CRPVb genomes differed at 238 (3%) nucleotide positions, whereas CRPVa4 and the prototype CRPV differed by only 5 nucleotides. The most variable region (7% nucleotide divergence) included the long regulatory region (LRR) and the E6 and E7 genes. A mutation in the stop codon resulted in an 8-amino-acid-longer CRPVb E4 protein, and a nucleotide deletion reduced the coding capacity of the E5 gene from 101 to 25 amino acids. In domestic rabbits homozygous for a specific haplotype of the DRA and DQA genes of the major histocompatibility complex, warts induced by CRPVb DNA or a chimeric genome containing the CRPVb LRR/E6/E7 region showed an early regression, whereas warts induced by CRPVa4 or a chimeric genome containing the CRPVa4 LRR/E6/E7 region persisted and evolved into carcinomas. In contrast, most CRPVa, CRPVb, and chimeric CRPV DNA-induced warts showed no early regression in rabbits homozygous for another DRA-DQA haplotype. Little, if any, viral replication is usually observed in domestic rabbit warts. When warts induced by CRPVa and CRPVb virions and DNA were compared, the number of cells positive for viral DNA or capsid antigens was found to be greater by 1 order of magnitude for specimens induced by CRPVb. Thus, both sequence variation in the LRR/E6/E7 region and the genetic constitution of the host influence the expression of the oncogenic potential of CRPV. Furthermore, intratype variation may overcome to some extent the host restriction of CRPV replication in domestic rabbits. PMID- 11044122 TI - Lentivirus vector gene expression during ES cell-derived hematopoietic development in vitro. AB - The murine embryonal stem (ES) cell virus (MESV) can express transgenes from the long terminal repeat (LTR) promoter/enhancer in undifferentiated ES cells, but expression is turned off upon differentiation to embryoid bodies (EBs) and hematopoietic cells in vitro. We examined whether a human immunodeficiency virus type 1-based lentivirus vector pseudotyped with the vesicular stomatitis virus G protein (VSV-G) could transduce ES cells efficiently and express the green fluorescent protein (GFP) transgene from an internal phosphoglycerate kinase (PGK) promoter throughout development to hematopoietic cells in vitro. An oncoretrovirus vector containing the MESV LTR and the GFP gene was used for comparison. Fluorescence-activated cell sorting analysis of transduced CCE ES cells showed 99.8 and 86.7% GPF-expressing ES cells in the VSV-G-pseudotyped lentivirus (multiplicity of infection [MOI] = 59)- and oncoretrovirus (MOI = 590) transduced cells, respectively. Therefore, VSV-G pseudotyping of lentiviral and oncoretrovirus vectors leads to efficient transduction of ES cells. Lentivirus vector integration was verified in the ES cell colonies by Southern blot analysis. When the transduced ES cells were differentiated in vitro, expression from the oncoretrovirus LTR was severely reduced or extinct in day 6 EBs and ES cell-derived hematopoietic colonies. In contrast, many lentivirus-transduced colonies, expressing the GFP gene in the undifferentiated state, continued to express the transgene throughout in vitro development to EBs at day 6, and many continued to express in cells derived from hematopoietic colonies. This experimental system can be used to analyze lentivirus vector design for optimal expression in hematopoietic cells and for gain-of-function experiments during ES cell development in vitro. PMID- 11044123 TI - Thogoto virus matrix protein is encoded by a spliced mRNA. AB - Thogoto virus (THOV) is a tick-transmitted orthomyxovirus with a segmented, negative-stranded RNA genome. In this study, we investigated the coding strategy of RNA segment 6 and found that it contains 956 nucleotides and codes for the matrix (M) protein. The full-length cDNA contains a single, long reading frame that lacks a stop codon but has coding capacity for a putative 35-kDa protein. In contrast, the M protein of THOV has an apparent molecular mass of 29 kDa as assessed by polyacrylamide gel electrophoresis. Therefore, we investigated the possibility of posttranscriptional processing of segment 6 transcripts by reverse transcription-PCR and identified a spliced mRNA that contains a stop codon and is translated into the 29-kDa M protein. Interestingly, the nontemplated UGA stop codon is generated by the splicing event itself. Thus, the unusual M coding strategy of THOV resembles that of Influenza C virus. PMID- 11044124 TI - Separable mechanisms of attachment and cell uptake during retrovirus infection. AB - In the absence of viral envelope gene expression, cells expressing human immunodeficiency virus type 1 (HIV-1) gag and pol, accessory HIV functions, and a vector genome RNA produce and secrete large amount of noninfectious virus-like particles (VLPs) into the conditioned medium. After partial purification, such HIV-1 VLPs can be made infectious in cell-free conditions in vitro by complex formation with lipofection reagents or with the G protein of vesicular stomatitis virus (VSV-G). The resulting in vitro-modified HIV-1 particles are able to infect nondividing cells. Infectivity of envelope-free HIV VLPs can also be induced by prior modification of target cells through exposure to partially purified VSV-G vesicles. Similarly, infection can be carried out by attachment of envelope-free noninfectious VLPs to unmodified cells followed by subsequent treatment of cells with VSV-G. We interpret these findings to indicate that interaction between a viral envelope and a cell surface receptor is not necessary for the initial virus binding to the cells but is required for subsequent cell entry and infection. PMID- 11044125 TI - Roles of Pr55(gag) and NCp7 in tRNA(3)(Lys) genomic placement and the initiation step of reverse transcription in human immunodeficiency virus type 1. AB - To study in vivo tRNA(3)(Lys) genomic placement and the initiation step of reverse transcription in human immunodeficiency virus type 1, total viral RNA isolated from either wild-type or protease-negative (PR(-)) virus was used as the source of primer tRNA(3)(Lys)/genomic RNA templates in an in vitro reverse transcription assay. At low dCTP concentrations, both the rate and extent of the first nucleotide incorporated into tRNA(3)(Lys), dCTP, were lower with PR(-) than with wild-type total viral RNA. Transient in vitro exposure of either type of primer/template RNA to NCp7 increased PR(-) dCTP incorporation to wild-type levels but did not change the level of wild-type dCTP incorporation. Exposure of either primer/template to Pr55(gag) had no effect on initiation. These results indicate that while Pr55(gag) is sufficient for tRNA(3)(Lys) placement onto the genome, exposure of this complex to mature NCp7 is required for optimum tRNA(3)(Lys) placement and initiation of reverse transcription. PMID- 11044126 TI - Rotavirus infection induces cytoskeleton disorganization in human intestinal epithelial cells: implication of an increase in intracellular calcium concentration. AB - Rotavirus infection is the most common cause of severe infantile gastroenteritis worldwide. In vivo, rotavirus exhibits a marked tropism for the differentiated enterocytes of the intestinal epithelium. In vitro, differentiated and undifferentiated intestinal cells can be infected. We observed that rotavirus infection of the human intestinal epithelial Caco-2 cells induces cytoskeleton alterations as a function of cell differentiation. The vimentin network disorganization detected in undifferentiated Caco-2 cells was not found in fully differentiated cells. In contrast, differentiated Caco-2 cells presented Ca(2+) dependent microtubule disassembly and Ca(2+)-independent cytokeratin 18 rearrangement, which both require viral replication. We propose that these structural alterations could represent the first manifestations of rotavirus infected enterocyte injury leading to functional perturbations and then to diarrhea. PMID- 11044127 TI - Molecular correlates of influenza A H5N1 virus pathogenesis in mice. AB - Highly pathogenic avian influenza A H5N1 viruses caused an outbreak of human respiratory illness in Hong Kong. Of 15 human H5N1 isolates characterized, nine displayed a high-, five a low-, and one an intermediate-pathogenicity phenotype in the BALB/c mouse model. Sequence analysis determined that five specific amino acids in four proteins correlated with pathogenicity in mice. Alone or in combination, these specific residues are the likely determinants of virulence of human H5N1 influenza viruses in this model. PMID- 11044128 TI - Development of a rubella virus vaccine expression vector: use of a picornavirus internal ribosome entry site increases stability of expression. AB - Rubella virus (RUB) is a small plus-strand RNA virus classified in the Rubivirus genus of the family Togaviridae. Live, attenuated RUB vaccines have been successfully used in vaccination programs for over 25 years, making RUB an attractive vaccine vector. In this study, such a vector was constructed using a recently developed RUB infectious cDNA clone (Robo). Using a standard strategy employed to produce expression and vaccine vectors with other togaviruses, the subgenomic promoter was duplicated to produce a recombinant construct (termed dsRobo) that expressed reporter genes such as chloramphenicol acetyltransferase and green fluorescent protein (GFP) under control of the second subgenomic promoter. However, expression of the reporter genes, as exemplified by GFP expression by dsRobo/GFP virus, was unstable during passaging, apparently due to homologous recombination between the subgenomic promoters leading to deletion of the GFP gene. To improve the stability of the vector, the internal ribosome entry site (IRES) of a picornavirus, encephalomyocarditis virus, was used instead of the second subgenomic promoter to eliminate homology. Construction was initiated by first replacing the subgenomic promoter in the parent Robo infectious clone with the IRES. Surprisingly, viable virus resulted; this virus did not synthesize a subgenomic RNA. The subgenomic promoter was then reintroduced in an orientation such that a single subgenomic RNA was produced, GFP was the initial gene on this RNA, while the RUB structural protein open reading frame was downstream and under control of the IRES element. GFP expression by this vector was significantly improved in comparison to dsRobo/GFP. This strategy should be applicable to increase the stability of other togavirus vectors. PMID- 11044129 TI - Replication of wild-type and mutant human cytomegalovirus in life-extended human diploid fibroblasts. AB - A cDNA encoding the catalytic subunit of human telomerase was used to generate life-extended derivatives of primary human diploid fibroblasts. The life-extended cells supported efficient human cytomegalovirus (HCMV) replication. A subclone of the life-extended cells was generated containing the HCMV UL82 gene and used to isolate and propagate a virus that exhibited a profound growth defect after infection at a low input multiplicity. PMID- 11044130 TI - The genomic RNA in Ty1 virus-like particles is dimeric. AB - The yeast retrotransposon Ty1 resembles retroviruses in a number of important respects but also shows several fundamental differences from them. We now report that, as in retroviruses, the genomic RNA in Ty1 virus-like particles is dimeric. The Ty1 dimers also resemble retroviral dimers in that they are stabilized during the proteolytic maturation of the particle. The stabilization of the dimer suggests that one of the cleavage products of TyA1 possesses nucleic acid chaperone activity. PMID- 11044131 TI - Concerted action of multiple cis-acting sequences is required for Rev dependence of late human immunodeficiency virus type 1 gene expression. AB - Based on the human immunodeficiency virus type 1 (HIV-1) gag gene, subgenomic reporter constructs have been established allowing the contributions of different cis-acting elements to the Rev dependency of late HIV-1 gene products to be determined. Modification of intragenic regulatory elements achieved by adapting the codon usage of the complete gene to highly expressed mammalian genes resulted in constitutive nuclear export allowing high levels of Gag expression independent from the Rev/Rev-responsive element system and irrespective of the absence or presence of the isolated major splice donor. Leptomycin B inhibitor studies revealed that the RNAs derived from the codon-optimized gag gene lacking AU-rich inhibitory elements are directed to a distinct, CRM1-independent, nuclear export pathway. PMID- 11044132 TI - Comparative analysis of translation efficiencies of hepatitis C virus 5' untranslated regions among intraindividual quasispecies present in chronic infection: opposite behaviors depending on cell type. AB - Hepatitis C virus (HCV) RNA translation initiation is dependent on the presence of an internal ribosome entry site (IRES) that is found mostly in its 5' untranslated region (5' UTR). While exhibiting the most highly conserved sequence within the genome, the 5' UTR accumulates small differences, which may be of biological and clinical importance. In this study, using a bicistronic dual luciferase expression system, we have examined the sequence of 5' UTRs from quasispecies characterized in the serum of a patient chronically infected with HCV genotype 1a and its corresponding translational activity. Sequence heterogeneity between IRES elements led to important changes in their translation efficiency both in vitro and in different cell cultures lines, implying that interactions of RNA with related transacting factors may vary according to cell type. These data suggest that variants occasionally carried by the serum prior to reinfection could be selected toward different compartments of the same infected organism, thus favoring the hypothesis of HCV multiple tropism. PMID- 11044133 TI - Porcine reproductive and respiratory syndrome virus: description of persistence in individual pigs upon experimental infection. AB - We studied the persistence of porcine reproductive and respiratory syndrome virus (PRRSV) in individual experimentally infected pigs, during a period of up to 150 days postinfection (dpi). The results of this study suggest that the persistence of PRRSV involves continuous viral replication but that it is not a true steady state persistent infection. The virus eventually clears the body and seems to do it in most of the animals by 150 dpi or shortly thereafter. High genetic stability was seen for several regions of the persistent PRRSV's genome, although some consistent mutations in the genes of envelope glycoproteins and M protein were also observed. PMID- 11044134 TI - Latent membrane protein 2A-mediated effects on the phosphatidylinositol 3 Kinase/Akt pathway. AB - Epstein-Barr virus (EBV) latent membrane protein 2A (LMP2A) is expressed on the membranes of B lymphocytes and blocks B-cell receptor (BCR) signaling in EBV transformed B lymphocytes in vitro. The phosphotyrosine motifs at positions 74 or 85 and 112 within the LMP2A amino-terminal domain are essential for the LMP2A mediated block of B-cell signal transduction. In vivo studies indicate that LMP2A allows B-cell survival in the absence of normal BCR signals. A possible role for Akt in the LMP2A-mediated B-cell survival was investigated. The protein kinase Akt is a crucial regulator of cell survival and is activated within B lymphocytes upon BCR cross-linking. LMP2A expression resulted in the constitutive phosphorylation of Akt, and this LMP2A effect is dependent on phosphatidylinositol 3-kinase activity. In addition, recruitment of Syk and Lyn protein tyrosine kinases (PTKs) to tyrosines 74 or 85 and 112, respectively, are critical for LMP2A-mediated Akt phosphorylation. However, the ability of LMP2A to mediate a survival phenotype downstream of Akt could not be detected in EBV negative Akata cells. This would indicate that LMP2A is not responsible for EBV dependent Burkitt's lymphoma cell survival. PMID- 11044135 TI - ATP binding and ATPase activities associated with recombinant rabbit hemorrhagic disease virus 2C-like polypeptide. AB - The carboxy-terminal region of the rabbit hemorrhagic disease virus p37 polyprotein cleavage product has been expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein. The recombinant GST-Delta2C protein showed in vitro ATP-binding and ATPase activities. Site-directed mutagenesis studies of the conserved residues G(522) and T(529) in motif A, D(566) and E(567) in motif B, and K(600) in motif C were also performed. These results provide the first experimental characterization of a 2C-like ATPase activity in a member of the Caliciviridae. PMID- 11044137 TI - Environmental threats to children's health: a challenge for pediatrics: 2000 Ambulatory Pediatric Association (APA) Presidential Address. PMID- 11044138 TI - George Armstrong Lecture 2000. PMID- 11044136 TI - Mechanisms for adaptation of simian immunodeficiency virus to replication in alveolar macrophages. AB - In contrast to the simian immunodeficiency virus SIVmac239, which replicates poorly in rhesus monkey alveolar macrophages, a variant with nine amino acid changes in envelope (SIVmac239/316E) replicates efficiently and to high titer in these same cells. We examined levels of viral DNA, RNA, antigen, and infectious virus to identify the nature of the block to SIVmac239 replication in these cells. Low levels of viral antigen (0.1 to 1.0 ng of p27 per ml) and infectious virus (100 to 1,000 infectious units per ml) were produced in the supernatant 1 to 4 days after SIVmac239 infection, but these levels did not increase subsequently. SIVmac239 DNA was synthesized in these macrophage cultures during the initial 24 h after infection, but the levels did not increase subsequently. Quantitation of the numbers of infectious cells in cultures over time and the results of experiments in which cells were reexposed to SIVmac239 after the initial exposure indicated that only a small proportion of cells were susceptible to SIVmac239 infection in these alveolar macrophage cultures and that the vast majority (>95%) of cells were refractory to SIVmac239 infection. In contrast to the results with SIVmac239, the levels of viral antigen, infectious virus, and viral DNA increased exponentially 2 to 7 days after infection by SIVmac239/316E, reaching levels greater than 100 ng of p27 per ml and 100,000 infectious units per ml. Since SIVmac239/316E has previously been described as a virus capable of infecting cells in a relatively CD4-independent fashion, we examined the levels of CD4 expression on the surface of fresh and cultured alveolar macrophages from rhesus monkeys. The levels of CD4 expression were extremely low, below the limit of detection by flow cytometry, on greater than 99% of the macrophages. CCR5(+) cells were profoundly depleted only from alveolar macrophage cultures infected with SIVmac239/316E. High concentrations of an antibody to CD4 delayed but did not block replication of SIVmac239/316E. The results suggest that the adaptation of SIVmac316 to efficient replication in alveolar macrophages results from its ability to infect these cells in a CD4-independent fashion or in a CD4-dependent fashion even at extremely low levels of surface CD4 expression. Since resident macrophages in brains and lungs of humans also express little or no CD4, our findings predict the presence of human immunodeficiency virus type 1 that is relatively CD4 independent in the lung and brain compartments of infected people. PMID- 11044140 TI - Self-reported physician practices for children with asthma: are national guidelines followed? AB - OBJECTIVE: To determine self-reported adherence to national asthma guidelines for children by primary care physicians in managed care; and, to analyze sources of variation in these practices by physician specialty and managed care practice type. DESIGN: A survey of 671 primary care physicians (pediatricians and family physicians) practicing in 3 geographically diverse managed care organizations (MCO). Domains of interest included asthma diagnosis, pharmacotherapy, patient education and follow-up, and indications for specialty referral. Item formats included self-reports of usual practice and responses to case vignettes. RESULTS: A total of 429 (64%) physicians returned surveys, 22 of whom did not meet criteria for inclusion in the analysis. Most respondents had both heard of (91%) and read (72%) the National Asthma Education and Prevention Program (NAEPP) guidelines. For diagnosis, 75% reported routine use of office peak flow measurement, but only 21% used spirometry routinely. Family physicians were more likely than pediatricians to use spirometry in diagnosis (odds ratio [OR] = 5.9), and less likely to recommend daily peak flow measurement (OR =.3). The median reported frequency of providing written care plans was only 50%. Though inhaled corticosteroids were deemed very safe or safe by 93%, almost half had specific concerns regarding at least 1 side effect, most commonly growth delay. Primary care physicians' criteria for referral to an asthma specialist differed from those of the NAEPP panel in choosing to manage more severe patients without asthma specialist input. Family physicians were more likely than pediatricians to refer a child after a single hospitalization, 2 to 3 emergency department visits, after 2 exacerbations, or if the child was <3 years old and required daily medications. Responses to vignettes showed generally appropriate initial use of antiinflammatory agents, but reluctance to increase the dose in response to continued symptoms, and less frequent follow-up than recommended by the NAEPP. CONCLUSION: Most physicians for children report having read and adopted NAEPP guideline recommendations for asthma treatment, including generally appropriate use of medications. Opportunities for improvement exist in specific areas such as the use of written care plans, optimizing antiinflammatory dosing, and providing routine follow-up. Although physicians show evidence of awareness of national guidelines and knowledge consistent with much of their content, additional work is required to promote the use of self-management tools in practice. PMID- 11044139 TI - Measuring the process of preventive service delivery in primary care practices for children. AB - BACKGROUND: Children may fall behind on preventive services because they do not receive needed services at the time of an office visit (a missed opportunity). However, methods are needed to measure problems in the care delivery process that lead to missed opportunities. We developed a method to examine the key steps in the preventive service delivery process and identify problems; we assessed the feasibility and validity of the method in primary care practices for children. METHODS: Using 3 data collection methods, we measured key steps in the process of preventive service delivery in primary care offices: a chart audit was used to measure each child's preventive service status before and after an office visit, a brief parent exit interview was used to assess preventive service delivery not documented in the chart, and a staff checklist was used to assess the role of nursing and other office staff. The feasibility of using this combination of measures to identify problems in the care delivery process was evaluated in 3 representative primary care practices (2 pediatric, 1 family practice) among children 5 years and younger. RESULTS: The measurement method was implemented in all 3 practices. The validity of the method was supported by its ability to detect differences among practices in the proportion of children eligible for immunizations and screening tests and in the proportion of children undergoing key steps in the process of preventive service delivery. The practice with the lowest proportion of children whose charts were screened for preventive services needs had the lowest performance of preventive services. CONCLUSIONS: It is possible to assess specific elements in the process of preventive service delivery in primary care practices. Use of this approach may help practices design and monitor interventions to improve the quality of preventive care delivery. PMID- 11044141 TI - Research and asthma: where do we go from here? PMID- 11044142 TI - Spirituality, religion, and pediatrics: intersecting worlds of healing. AB - Religious practices such as prayer represent the most prevalent complementary and alternative therapies in the United States. However, biomedicine has sometimes viewed faith and related religious worldviews as relevant only when they obstruct implementation of scientifically sound biomedical care. Recent efforts to arrive at a new synthesis raise challenges for pediatricians. This article reviews theories of child faith development, and models of child spirituality from different disciplinary perspectives. It provides sources illustrating how spirituality and religion may inform children's lives; play a part in children's moral formation, socialization, and induction into a sacred worldview; and provide the child with inner resources. It also suggests some of the positive and negative effects of spiritual and religious engagement. Second, this article examines aspects of spirituality and religion that parents may bring to bear in relation to their children's health. Third, this article addresses the spiritual and/or religious identity of the provider. These topics are discussed in the context of cultural competence and the related importance of religious diversity. The authors suggest 1) some approaches for appropriate inclusion of spirituality in clinical practice, 2) challenges for medical education, and 3) areas requiring further research. PMID- 11044143 TI - Health care needs of children in the foster care system. AB - Nearly 750 000 children are currently in foster care in the United States. Recent trends in foster care include reliance on extended family members to care for children in kinship care placements, increased efforts to reduce the length of placement, acceleration of termination of parental rights proceedings, and emphasis on adoption. It is not clear what impact welfare reform may have on the number of children who may require foster care placement. Although most children enter foster care with medical, mental health, or developmental problems, many do not receive adequate or appropriate care while in placement. Psychological and emotional problems, in particular, may worsen rather than improve. Multiple barriers to adequate health care for this population exist. Health care practitioners can help to improve the health and well-being of children in foster care by performing timely and thorough admission evaluations, providing continuity of care, and playing an active advocacy role. Potential areas for health services research include study of the impact of different models of health care delivery, the role of a medical home in providing continuity of care, the perception of the foster care experience by the child, children's adjustment to foster care, and foster parent education on health outcomes. PMID- 11044144 TI - Impact of appointment reminders on vaccination coverage at an urban clinic. AB - OBJECTIVE: To test if appointment reminders blinded to immunization status improve kept-appointment and vaccination coverage rates. Design. Controlled trial. SETTING: Pediatric clinic serving a low-income community in New York City. INTERVENTION: Children ages 4 through 18 months (n = 1273) scheduled sequentially for clinic appointments were systematically assigned to 1 of 4 study groups: control (n = 346); postcard (n = 314); telephone call (n = 307); and postcard and telephone call (n = 306). OUTCOME MEASURES: Kept-appointment and vaccination coverage rates. RESULTS: Children assigned to the postcard and telephone group were 1.75 times more likely to keep their appointments than controls (95% confidence interval [CI] = 1.2, 2.5). Children who actually received the postcard and telephone reminders were 2.3 times more likely to keep an appointment than controls (95% CI = 1.4, 3.7). Children who kept appointments were 2.3 times more likely to be up-to-date with their immunizations (95% CI = 1.7, 3.2). The reminders selectively increased vaccination coverage for the subgroup of children who were not up-to-date before the appointment (chi(2) = 11.2). The cost of the reminders was $.67 for the postcard and $1.58 for the postcard and telephone. Assuming 5000 visits per year and $100 reimbursement per visit, the return on each dollar invested was $10 for the postcard and $7.28 for the postcard and telephone reminder. CONCLUSIONS: Appointment reminders blind to immunization status are a practical and cost-effective strategy to increase kept-appointment rates for all children, and, through this mechanism, reach and vaccinate children who are not up-to-date.appointment reminder, vaccination coverage. PMID- 11044145 TI - The association between hands-on instruction and proper child safety seat installation. AB - OBJECTIVE: To determine if hands-on instruction in child safety seat (CSS) installation decreases the number of errors in installation. DESIGN: Cross sectional study. SETTING: Primary care offices, emergency department, CSS checkpoint. PARTICIPANTS: Parents of children <2 years old receiving medical care or attending a CSS check. MAIN OUTCOME MEASURE: Errors in CSS use. Results. Only 6.4% of parents had a correctly installed CSS. Hands-on instruction was associated with fewer errors in seat installation. Increased parent age, completion of college, and having private insurance were also associated with fewer errors in CSS placement. The majority of parents learned to install seats from reading the manual, from friends and relatives, and from figuring it out on their own. CONCLUSIONS: Errors in CSS installation are a significant problem. Hands-on instruction decreases the numbers of errors in CSS installation. However, few parents receive hands-on instruction from experts in CSS installation. Increases in correct CSS use could result from hands-on education by trained professionals. PMID- 11044146 TI - Uninsured children with psychosocial problems: primary care management. AB - OBJECTIVE: Nearly 14% of children in the United States are uninsured. We compared the prevalence of psychosocial problems and mental health services received by insured and uninsured children in primary care practices. METHODS: The Child Behavior Study was a cohort study conducted by Pediatric Research in Office Settings and the Ambulatory Sentinel Practice Network. Four hundred one primary care clinicians enrolled an average sample of 55 consecutive children (4-15 years old) per clinician. RESULTS: Of the 13 401 visits to clinicians with 3 or more uninsured patients, 12 518 were by insured children (93.4%) and 883 were by uninsured children (6. 6%). A higher percentage of adolescents, Hispanic children, those with unmarried parents, and those with less educated parents were uninsured. According to clinicians, uninsured children and insured children had similar rates of psychosocial problems (19%) and severe psychosocial problems (2%). For children with a clinician-identified psychosocial problem, we found no differences in clinician-reported counseling, medication use, or referral to mental health professionals. CONCLUSIONS: Among children served in primary care practices, uninsured children have similar prevalence of clinician-identified psychosocial and mental health problems compared with insured children. Within their practices, clinicians managed uninsured children much the same way as insured children. psychosocial problems, uninsured children, pediatrics, family medicine, primary care. PMID- 11044147 TI - Effect of a reduced postpartum length of stay program on primary care services use by mothers and infants. AB - OBJECTIVE: To determine the impact of reduced postpartum length of stay (LOS) on primary care services use. METHODS. DESIGN: Retrospective quasiexperimental study, comparing 3 periods before and 1 period after introducing an intervention and adjusting for time trends. SETTING: A managed care plan. INTERVENTION: A reduced obstetrical LOS program (ROLOS), offering enhanced education and services. PARTICIPANTS: mother-infant dyads, delivered during 4 time periods: February through May 1992, 1993, and 1994, before ROLOS, and 1995, while ROLOS was in effect. INDEPENDENT MEASURES: Pre-ROLOS or the post-ROLOS year. OUTCOME MEASURES: Telephone calls, visits, and urgent care events during the first 3 weeks postpartum summed as total utilization events. RESULTS: Before ROLOS, LOS decreased gradually (from 51.6 to 44.3 hours) and after, sharply to 36.5 hours. Although primary care use did not increase before ROLOS, utilization for dyads increased during ROLOS. Before ROLOS, there were between 2.37 and 2.72 utilization events per day; after, there were 4.60. Well-child visits increased slightly to.98 visits per dyad, but urgent visits did not. CONCLUSION: This program resulted in shortened stays and more primary care use. There was no increase in infant urgent primary care utilization. Early discharge programs that incorporate and reimburse for enhanced ambulatory services may be safe for infants; these findings should not be extrapolated to mandatory reduced LOS initiatives without enhancement of care. PMID- 11044148 TI - Variation in hospital discharges for ambulatory care-sensitive conditions among children. AB - OBJECTIVE: Ambulatory Care-Sensitive Conditions (ACSCs), conditions for which ambulatory care may reduce, though not eliminate, the need for hospital admission, have been used as an index of adequate primary care. However, few studies of ACSC have focused on children. We estimated national hospitalization rates for ACSC among children and examined the behavior of the index between subgroups of children. METHODS: We used data from the 1990-1995 National Hospital Discharge Surveys (NHDS), the US census, and the National Health Interview Survey (NHIS) to calculate hospital discharge rates. Rates were estimated as the number of condition-specific hospital discharges from the NHDS divided by the population at risk, as estimated from the US census and NHIS. RESULTS: Predictably, ACSC hospitalization rates were significantly higher among children who were younger, black, had Medicaid insurance, and lived in poorer areas compared with their counterparts. However, the relationship between ACSCs and income and the distributions of conditions within the index varied significantly between children. CONCLUSIONS: ACSCs may indicate disparities in access and utilization of health care, however, the differing behavior of the index between subgroups suggests that inferences from examining rates of ACSCs may not be comparable for all children.ambulatory care-sensitive conditions, hospitalization rates. PMID- 11044150 TI - Medical management of bowel obstruction. PMID- 11044149 TI - Exposure to violence among urban school-aged children: is it only on television? AB - OBJECTIVE: To measure exposure to different types of violence among school-aged children in a primary care setting. DESIGN: Child interviews using an instrument measuring 4 types of exposure (direct victimization, witnessing, hearing reports, media). Violent acts measured include being beaten up, chased/threatened, robbed/mugged, stabbed/shot, killed. SETTING: Pediatric primary care clinic of large urban hospital. PATIENTS: Convenience sample of 175 children 9-12 years old and their mothers. A total of 53% of the children were boys, 55% were Hispanic, and 40% received public assistance. RESULTS: All children had been exposed to media violence. A total of 97% (170/175) had been exposed to more direct forms of violence; 77% had witnessed violence involving strangers; 49% had witnessed violence involving familiar persons; 49% had been direct victims; and 31% had witnessed someone being shot, stabbed, or killed. Exposure to violence was significantly associated with being male. CONCLUSION: Most school-aged children who visited a pediatric primary care clinic of a large urban hospital had directly experienced violence as witnesses and/or victims. PMID- 11044151 TI - Politics in surgical publishing. PMID- 11044152 TI - COPE: committee on publication ethics PMID- 11044153 TI - Anatomy of Denonvilliers' fascia and pelvic nerves, impotence, and implications for the colorectal surgeon. AB - BACKGROUND: The development and anatomy of Denonvilliers' fascia have been controversial for many years and confusion exists about its operative appearance. Better appreciation of this poorly understood anatomy, and its significance for impotence after rectal dissection, may lead to further functional improvements in pelvic surgery. METHOD: A literature review of the embryology and anatomy of Denonvilliers' fascia and impotence after pelvic rectal surgery was undertaken. RESULTS: Denonvilliers' fascia has no macroscopically discernible layers. The so called posterior layer refers to the fascia propria of the rectum. The incidence of erectile and ejaculatory dysfunction after rectal excision is high in older patients, and when performed for rectal cancer. There is no consensus about the relationship of Denonvilliers' fascia to the plane of anterior dissection for rectal cancer. CONCLUSION: Colorectal surgeons should focus on the important anatomy between the rectum and the prostate to improve functional outcomes after rectal excision. A classification of the available anterior dissection planes is proposed. Surgeons should be encouraged to document the plane used as well as outcome in terms of sexual function. PMID- 11044154 TI - Vascular access for haemodialysis. AB - BACKGROUND: The recent expansion of renal replacement therapy programmes has been associated with an increase in the number and complexity of patients requiring permanent vascular access. The introduction of strategies designed to maximize secondary access patency is, therefore, increasingly important as a means of prolonging patient survival on dialysis, reducing morbidity and reducing the escalating cost of such programmes. METHODS: A review of the current literature on the planning of vascular access, access surveillance methods and treatment of the most common complications was performed. RESULTS: Multidisciplinary vascular access planning, increased use of preoperative imaging and the preferential use of autogeneous vein are essential to obtain the best long-term results. While vascular access surveillance, in particular protocols involving direct measurement of access flow, enables the prospective detection and treatment of venous stenosis, the precise indications for treating venous stenosis remain unclear. Surgical revision remains the gold standard for the treatment of failing arteriovenous fistulas, but recent advances in interventional radiological techniques along with the suitability of arteriovenous fistulas for percutaneous intervention may offer an effective alternative. The effect of both these interventions on access patency requires comparison in a randomized trial. CONCLUSION: The introduction of strategies to improve access patency rates will change vascular access surgical practice away from the construction of new fistulas towards an increase in outpatient percutaneous intervention and surgical revisional procedures. The role of surgical interventions requires clearer definition. PMID- 11044156 TI - DIGEST: surgery today PMID- 11044155 TI - Factors predictive of outcome after surgery for faecal incontinence. AB - BACKGROUND: Surgical treatment of faecal incontinence may be categorized into procedures that either repair or augment the native sphincter mechanism or, alternatively, require construction of a neosphincter using either autologous tissue or an artificial device. METHODS: This article reviews the currently available surgical options for the treatment of faecal incontinence, discusses factors predictive of outcome, and includes an algorithm for treatment. RESULTS AND CONCLUSION: Procedures such as postanal repair, direct sphincter repair and reefing are seldom used. Overlapping repair has become the operation of choice in incontinent patients with isolated anterior defects in the external anal sphincter muscle, particularly in postobstetric trauma. Pudendal neuropathy seems to be a predictive factor of success, although this is not universally accepted. Total pelvic floor repair has been offered as a recent alternative. Neosphincter procedures include a gluteoplasty, non-stimulated and stimulated unilateral or bilateral graciloplasty and artificial bowel sphincter. The success and morbidity rates with the stimulated graciloplasty and artificial bowel sphincter appear similar. The newest alternative, sacral nerve stimulation, seems promising. In the final analysis, case selection and surgical judgement are probably the most important factors influencing the success of surgery for faecal incontinence. Presented as the Edinburgh Royal College of Surgeons invited lecture to the Association of Coloproctology of Great Britain and Ireland, Southport, UK, June 1999 PMID- 11044157 TI - Hypertonic saline attenuates end-organ damage in an experimental model of acute pancreatitis. AB - BACKGROUND: Hypertonic saline (HTS) has been noted previously to reduce neutrophil activation. The aim of this study was to elucidate the effect of hypertonic resuscitation on the development of end-organ damage in an animal model of pancreatitis. METHODS: Pancreatitis was induced in Sprague-Dawley rats by intraperitoneal injection of 20 per cent L-arginine. Animals were randomized into four groups (each n = 8): controls; pancreatitis without intervention; pancreatitis plus intravenous resuscitation with normal saline (0.9 per cent sodium chloride 2 ml/kg) at 24 and 48 h; or HTS (7.5 per cent sodium chloride 2 ml/kg) at these time points. Pulmonary endothelial leakage was assessed by measurement of lung wet : dry ratios, bronchoalveolar lavage protein and myeloperoxidase activity. RESULTS: Animals that received HTS showed less pancreatic damage than those resuscitated with normal saline (1.0 versus 3.0; P = 0.04). Lung injury scores were also significantly diminished in the HTS group (1.0 versus 3.5; P = 0.03). Pulmonary neutrophil sequestration (myeloperoxidase activity 1.80 units/g) and increased endothelial permeability (bronchoalveolar lavage protein content 1287 microgram/ml) were evident in animals resuscitated with normal saline compared with HTS (1.22 units/g and 277 microgram/ml respectively; P < 0.02). CONCLUSION: HTS resuscitation results in a significant attenuation of end-organ injury following a systemic inflammatory response to severe pancreatitis. PMID- 11044158 TI - Chronic dysphagia following laparoscopic fundoplication. AB - BACKGROUND: Many surgeons practise tailored laparoscopic antireflux surgery in an attempt to prevent postoperative dysphagia. The aim of this study was to determine the effect of 360 degrees fundoplication (Nissen) or 270 degrees fundoplication (Toupet), and the influence of abnormal oesophageal peristalsis, upon postoperative dysphagia. METHODS: This was a cohort study from three tertiary referral centres, using dysphagia before laparoscopic fundoplication and 1 year after operation as the main outcome variable. Preoperative oesophageal manometry was performed on all patients. RESULTS: Some 761 patients underwent Nissen and 85 underwent Toupet fundoplication. Only 2 per cent reported severe postoperative dysphagia. There was a significant selection bias towards the Toupet operation for patients with abnormal oesophageal motility (P < 0.001). For patients whose oesophageal manometric findings were normal there was a significant improvement in dysphagia after Nissen fundoplication (P = 0.02), and no significant change following Toupet fundoplication. There was no significant change in the rate of dysphagia following either method of fundoplication amongst other subgroups in which oesophageal manometry was stratified as non-specific motor disorder, low-amplitude peristalsis, or aperistalsis. CONCLUSION: A tailored approach to the degree of fundoplication is unnecessary as patients with dysmotility suffer no more dysphagia after full laparoscopic Nissen fundoplication than those who have a partial Toupet wrap. PMID- 11044159 TI - Interleukin 10-deficient colitis: new similarities to human inflammatory bowel disease. AB - BACKGROUND: Interleukin (IL) 10 is a potent anti-inflammatory cytokine. Disruption of the IL-10 gene in C57/Black6 mice results in enterocolitis in the presence of intestinal bacteria. This study investigated gut mucosal barrier function sequentially during the development of colitis in this model. METHODS: Animals were bred in specific pathogen-free conditions and transferred to conventional housing at 4 weeks. Mice were evaluated at 6, 8, 10, 12, 14 and 15 weeks of age. Barrier function was assessed by measuring intestinal permeability and antibody response to systemic endotoxaemia (antibody to the core glycolipid region of lipopolysaccharide; EndoCAb). Colons were harvested and a histological injury score (HIS) was calculated. RESULTS: The HIS increased progressively until 12 weeks, with an associated increase in intestinal permeability, and immunoglobulin (Ig) M and IgG EndoCAb. The HIS correlated positively with both intestinal permeability and IgM and IgG EndoCAb. Intestinal permeability showed a positive correlation with EndoCAb. CONCLUSION: IL-10 knockout mice develop colitis with an associated disturbance in gut mucosal barrier function, as measured by increased permeability and endotoxaemia. The colitis found in the IL 10 knockout mouse shares these histological, physiological and biochemical features with human inflammatory bowel disease and is therefore suitable for therapeutic trials. A measure of endotoxaemia correlated directly with intestinal permeability in this model. PMID- 11044160 TI - Randomized clinical trial of sutured versus stapled closed haemorrhoidectomy. AB - BACKGROUND: Stapled haemorrhoidectomy does not involve dissection, with its attendant potential morbidity, required to perform closed haemorrhoidectomy. This study compared haemorrhoidectomy with (sutured) and without (stapled) preliminary dissection. METHODS: Forty patients with prolapsed symptomatic haemorrhoids were randomly assigned to sutured (n = 20) or stapled (n = 20) haemorrhoidectomy. Preoperative assessment was by proctoscopy, sigmoidoscopy and anal manometry. Stapled and diathermy haemorrhoidectomies with wound suture were performed, and excised tissue was examined histologically. Pain scores, complications, wound healing and patient satisfaction were recorded. Follow-up was weekly for 4 weeks, and at 3 and 6 months; anal manometry was repeated at the last two visits. RESULTS: Postoperative resting and squeeze pressures were reduced by the stapled method at 3 months (P = 0.02 and P = 0.03 respectively), returning to baseline by 6 months. Stapled haemorrhoidectomy was quicker but initial access into the anus was hampered by the bulky stapler. Isolated muscle fibres were identified equally in both groups, but incontinence did not occur. The stapled technique resulted in less postoperative pain (P = 0.04), a greater degree of satisfaction (P = 0.01) and faster wound healing (P < 0.001), but was more expensive. There was no significant difference in complications. CONCLUSION: Despite the higher cost and difficult access, stapled haemorhoidectomy results in less postoperative pain, faster wound healing and greater patient satisfaction than the sutured technique. PMID- 11044161 TI - Causes of re-recurrence after polytetrafluoroethylene patch saphenoplasty for recurrent varicose veins. AB - BACKGROUND: The aim of this study was to determine whether a polytetrafluoroethylene (PTFE) patch sutured over the religated saphenofemoral junction could reduce the rate of recurrence after operation for recurrent varicose veins. METHODS: Fifty patients who had surgery for recurrent long saphenous incompetence (81 legs had a small PTFE patch sutured over the religated saphenofemoral junction. There were no major complications following surgery. Three patients had a wound infection or delayed healing. All patients were invited for clinical examination and duplex imaging at a median of 19 (range 6 39) months after operation. RESULTS: Some 38 of 43 patients (70 legs) remained satisfied with the results of surgery; 16 (23 per cent) of 70 legs had visible veins on inspection and eight of these (11 per cent) involved symptomatic recurrence. Duplex imaging showed that recurrence was due to saphenofemoral junction incompetence in ten legs; two appeared to have a major groin connection but the other eight appeared to have neovascularization. Other causes were thigh perforator reflux (three legs) and cross-groin collaterals (three). Eleven of the 16 legs with recurrence had varicography but in two the procedure was a technical failure. Two legs had evidence of a significant connection (more than 3 mm) and two a minor connection (less than 3 mm) to the femoral vein at the level of the PTFE patch, but in the remainder recurrence was due to upper thigh perforating veins. There was good concordance between duplex imaging and varicography. CONCLUSION: PTFE patch saphenoplasty appears to be safe. Although these are early results, the technique seems potentially as effective as other barrier methods that have been investigated; in ten legs (12 per cent) recurrence was attributed to failure at the level of the PTFE patch. PMID- 11044162 TI - Short-term results of femoropopliteal subintimal angioplasty. AB - BACKGROUND: Subintimal angioplasty may be more successful than conventional (intraluminal) angioplasty for treatment of long femoropopliteal occlusions. This study assessed the clinical and haemodynamic outcome of subintimal angioplasty. METHODS: All patients with femoropopliteal occlusions treated by subintimal angioplasty over a 3-year period at two centres were reviewed. Clinical assessment and colour duplex imaging were carried out. RESULTS: Sixty-nine procedures were performed in 33 men and 33 women of median age 74 (range 47-92) years. Indications for treatment were intermittent claudication in 26 (38 per cent) and critical limb ischaemia in 43 (62 per cent). Median occlusion length was 10 (range 2-50) cm. Primary technical success was achieved in 51 occlusions (74 per cent). There were 11 complications (16 per cent); the majority were minor but surgical intervention was required in two patients (3 per cent). At 6 months the cumulative symptomatic and haemodynamic primary patency rates were 60 and 51 per cent respectively, analysed on an intention-to-treat basis. The symptomatic and haemodynamic patency rates for technically successful procedures were 80 and 77 per cent respectively. CONCLUSION: In this series the short-term clinical success of subintimal angioplasty was poor because of a high incidence of reocclusion and restenosis, despite a relatively high initial technical success rate. PMID- 11044163 TI - Multicentre, randomized clinical trial of primary versus secondary sigmoid resection in generalized peritonitis complicating sigmoid diverticulitis. AB - BACKGROUND: The best way to manage generalized peritonitis complicating sigmoid diverticulitis is controversial. This randomized clinical trial involved a comparison of primary resection and suture, drainage with proximal colostomy followed by secondary resection. METHODS: From January 1989 to December 1996, 105 patients of mean(s.d.) age 66(14) (range 32-91) years were randomized to undergo primary or secondary resection. The main endpoint was occurrence of generalized or localized postoperative peritonitis. The Mannheim Peritonitis Index score was calculated for each patient to check for comparability of groups. RESULTS: Postoperative peritonitis occurred less often after primary than secondary resection whether considering the first procedure only (one of 55 patients versus ten of 48; P < 0.01) or all procedures (one of 55 versus 12 of 48; P < 0.001). Likewise, early reoperation was performed less often following primary resection than secondary resection (two of 55 versus nine of 48 (P < 0.02) and two versus 11 (P < 0.01)), leading to a shorter median first hospital stay for patients having primary resection (15 days) than for those undergoing secondary resection (24 days) (P < 0.05). The mortality rate did not differ significantly with regard to operative policy (primary resection 24 per cent versus secondary resection 19 per cent) or type of peritonitis (faeculent 27 per cent versus purulent 19 per cent). No patient died following a second or third procedure. CONCLUSION: Primary resection is superior to secondary resection in the treatment of generalized peritonitis complicating sigmoid diverticulitis because of significantly less postoperative peritonitis, fewer reoperations and shorter hospital stay. PMID- 11044164 TI - Randomized controlled trial of the effect of early enteral nutrition on markers of the inflammatory response in predicted severe acute pancreatitis. AB - BACKGROUND: Recent evidence suggests that intestinal dysfunction has a role in sustaining the systemic inflammatory response in acute pancreatitis and may be ameliorated by the introduction of enteral nutrition. This study therefore assessed the effect of early enteral nutrition on the systemic inflammatory response in patients with prognostically severe acute pancreatitis. METHODS: Patients with prognostically severe acute pancreatitis within 72 h of disease onset were randomized to receive either enteral nutrition or conventional therapy consisting of a nil-by-mouth regimen. Serum interleukin (IL) 6, soluble tumour necrosis factor receptor I (sTNFRI) and C-reactive protein (CRP) were used as markers of the inflammatory response. Intestinal function was assessed using a differential sugar permeability technique. RESULTS: Of 27 patients, 13 received enteral nutrition. A median of 21 (range 0-100) per cent of calorific requirements was delivered over the first 4 days by enteral nutrition. There were no significant complications of enteral nutrition. The introduction of enteral nutrition did not affect the serum concentrations of IL-6 (P = 0.28), sTNFRI (P = 0.53) or CRP (P = 0.62) over the first 4 days of the study. Although there were no significant differences in intestinal permeability between the two patient groups at admission (chi2 = 2.33, d.f. = 1, P = 0.13), by day 4 abnormal intestinal permeability occurred more frequently in patients receiving enteral nutrition (chi2 = 4.94, d.f. = 1, P = 0.03) CONCLUSION: Early enteral nutrition did not ameliorate the inflammatory response in patients with prognostically severe acute pancreatitis. Furthermore, it did not have a beneficial effect on intestinal permeability. Presented in part to the Pancreatic Society of Great Britain and Ireland in Leeds, UK, November 1998 and at Digestive Disease Week in Orlando, Florida, USA, May 1999 PMID- 11044165 TI - Preventive effect of preoperative portal vein ligation on endotoxin-induced hepatic failure in hepatectomized rats is associated with reduced tumour necrosis factor alpha production. AB - BACKGROUND: Preoperative portal vein embolization successfully reduces the incidence of postoperative hepatic failure in which endotoxin is postulated to be involved. To identify the mechanism of this preventive effect, the relationship of endotoxin-induced liver injury with tumour necrosis factor (TNF) alpha and nitric oxide production in the peripheral blood, liver and spleen of rats subjected to preoperative portal vein branch ligation (PVL) was compared with that in rats undergoing sham operation. METHODS: Rats with PVL and those that underwent sham operation were subjected to resection of ligated liver lobes (PVL Hx rats) and two-thirds hepatectomy (noPVL-Hx rats) respectively at day 5, followed by intravenous administration of endotoxin 200 microgram/kg body-weight at day 7. At various time intervals after endotoxin injection, the peripheral blood, liver and spleen tissues were harvested and analysed for TNF-alpha and nitric oxide production. RESULTS: The survival rates of noPVL-Hx and PVL-Hx rats at 48 h after endotoxin administration were 40 and 100 per cent respectively. The former rats showed more extensive liver injury as represented by higher serum aminotransferase and hyaluronate levels than the latter. Plasma concentrations of TNF-alpha at 1.5 h after endotoxin treatment were significantly higher in noPVL Hx rats (mean(s.e.m.) 22 125(2175) pg/ml; n = 6) than PVL-Hx rats (8344(4076) pg/ml; n = 6) (P < 0.01). Consistent with this, expression of TNF-alpha messenger RNA in the liver and spleen was suppressed in PVL-Hx rats. In two-thirds hepatectomized rats, plasma TNF-alpha concentrations after endotoxin administration at 1, 2 and 3 days (14 350(2186), 26 375(2478) and 23 000(3745) pg/ml respectively; n = 6 each) were significantly higher than that before operation (9067(1559) pg/ml; n = 6) (P < 0.05), whereas those at 5 and 7 days (10 102(3616) and 8580(1427) pg/ml respectively; n = 6 each) showed no significant increase. Furthermore, nitric oxide production in peripheral blood and liver was suppressed by preoperative PVL. CONCLUSION: Prevention of endotoxin-induced liver failure by preoperative PVL is associated with reduced production of TNF-alpha in the later phase of liver regeneration. PMID- 11044166 TI - Localization of matrix metalloproteinase 2 within the aneurysmal and normal aortic wall. AB - BACKGROUND: Current research has shed new light on the role of matrix metalloproteinase (MMP) 2 in the development of abdominal aortic aneurysms (AAAs). MMP-2 is a major protease in the wall of small aneurysms and is produced at increased levels by smooth muscle cells derived from AAAs compared with normal controls. In vivo, MMP-2 is produced as an inactive proenzyme that is activated predominantly by the cell membrane-bound enzyme, membrane type 1 matrix metalloproteinase (MT1-MMP). This study investigated the production of the MMP-2 MT1-MMP-tissue inhibitor of metalloproteinases (TIMP) 2 system within the wall of aortic aneurysms and in age-matched control arterial tissue. METHODS: Arterial tissue from four patients with aortic aneurysms and four age-matched aortic samples was examined for the production and expression of MMP-2, TIMP-2 and MT1 MMP protein using immunohistochemistry, in situ hybridization and in situ zymography. RESULTS: All components of the MMP-2-TIMP-2-MT1-MMP enzyme system were detected in the arterial wall of both aneurysm and control samples, specifically in the medial tissue. The enzymes co-localized with medial smooth muscle cells. Gelatinolytic activity was localized to elastin fibres in normal and aneurysmal aorta. CONCLUSION: The presence of MT1-MMP within the media of arterial tissue suggests a powerful pathway for the activation of MMP-2. The localization of the MMP-2-TIMP-2-MT1-MMP enzyme system to the medial layer of the arterial wall gives support to the concept that this system may play an aetiological role in the pathogenesis of AAAs. PMID- 11044167 TI - Factors influencing the functional outcome of restorative proctocolectomy in ulcerative colitis. AB - BACKGROUND: Restorative proctocolectomy is considered to be the procedure of choice in the operative treatment of ulcerative colitis. The aim of this study was to evaluate the functional outcome following operation and to identify possible predictive factors. METHODS: Some 168 patients (median age 32 years, 102 men) with ulcerative colitis underwent restorative proctocolectomy. The functional outcome was evaluated by a symptom index created from a questionnaire at a median of 29 (13-123) months of follow-up. The records of these patients were reviewed, and preoperative, peroperative and postoperative variables were registered and related to outcome. RESULTS: The response rate to the questionnaire was 155 (92 per cent) of 168. The symptom index was related to patients' overall assessment of outcome. In spite of a perceived good result many patients experienced a number of symptoms. Age over 50 years (P < 0.01), presence of extraintestinal manifestations (P < 0.05) and late complications, such as anastomotic stricture (P < 0.05), pouchitis (P < 0.01) and anal pain (P < 0.05), were related to a less favourable outcome. CONCLUSION: While preoperative data may help in selecting patients suitable for restorative proctocolectomy, prevention of late complications seems most important in improving the functional outcome. PMID- 11044168 TI - Raised levels of plasma big endothelin 1 in patients with colorectal cancer. AB - BACKGROUND: The aim was to assess the role of plasma Big Endothelin (ET) 1 levels as a marker of disease presence and stage in colorectal adenocarcinoma. METHODS: Big ET-1 was measured in the plasma of 37 patients with colorectal cancer. Preoperative systemic plasma levels of Big ET-1 in patients with cancer were compared with levels in 20 age- and sex-matched controls. Portal plasma samples were collected at operation in addition to peripheral venous samples. Immunohistochemical staining for Big ET-1 was performed on a selection of primary tumour specimens and liver metastases. RESULTS: Median (range) preoperative systemic plasma levels of Big ET-1 were significantly higher in patients with cancer than in controls (1.0 (0.3-9.7) versus 0.2 (0.0-6.0) fmol/ml; P = 0.0001). Intraoperative portal plasma levels of Big ET-1 were significantly higher in patients with Dukes' 'D' disease than in patients with Dukes' A, B and C disease (2.1 (1.4-10.0) versus 1.2 (0.3-6.6) fmol/ml; P = 0. 01). Similarly, systemic plasma levels were significantly higher in patients with Dukes' 'D' disease than in those with localized disease (1.9 (1.2-9.7) versus 1.2 (0.2-8.3) fmol/ml; P = 0.01). The presence of microvascular invasion in the tumour specimens was associated with a significantly raised portal plasma level of Big ET-1 (1.6 (1.5 2.1) versus 1.1 (0.8-1.3) fmol/ml; P = 0.04). Immunohistochemistry localized Big ET-1 to the cancer epithelial cells. CONCLUSION: The plasma level of Big ET-1 is significantly raised in patients with colorectal cancer. Patients with liver metastases have significantly higher levels than those with localized disease. PMID- 11044169 TI - Clinical presentation of the Pseudomyxoma peritonei syndrome. AB - BACKGROUND: Pseudomyxoma peritonei syndrome is characterized by a gradual expansion of mucoid tumour and fluid at specific sites within abdominopelvic regions as a result of a perforated appendiceal adenoma. The aim was to analyse the presenting symptoms and signs of patients with this condition. METHODS: Of 410 patients with appendiceal tumours 217 had the diagnosis of pseudomyxoma peritonei syndrome with histological confirmation. A retrospective review of the clinical characteristics that determine presentation was performed. RESULTS: Overall, suspected acute appendicitis was the most common presentation (27 per cent). For women the diagnosis was most commonly made while being evaluated for an ovarian mass (39 per cent). Increasing abdominal girth was the second most common presentation overall (23 per cent). Thirty patients (14 per cent) presented with new-onset hernia, of which the majority (26) were inguinal hernias. CONCLUSION: Consideration of appendicitis, increased abdominal girth, ovarian mass and new-onset hernia as caused by this syndrome may facilitate diagnosis and definitive treatment. PMID- 11044170 TI - Recurrent sigmoid volvulus treated by percutaneous endoscopic colostomy. PMID- 11044171 TI - Effect of surgical subspecialization on breast cancer outcome. AB - BACKGROUND: There is increasing pressure for specialization of medical services. The effect of surgical specialization on the outcome of breast cancer in Bedford has been assessed. METHODS: The Bedford Breast Cancer Registry, which contains prospective diagnostic, treatment and follow-up data on all breast cancers treated in North Bedfordshire, was analysed to compare breast cancer outcome between 1990-1992 and 1993-1996, that is before and after the advent of surgical subspecialization. All 784 patients were analysed, including patients with metastases (4 per cent) and those treated by tamoxifen alone (8 per cent). Outcome was compared in terms of disease-free survival (DFS), locoregional and all (locoregional and metastases) recurrence rates assessed by Cox proportional hazard and Kaplan-Meier analyses. RESULTS: Overall DFS was 75 per cent and the locoregional recurrence rate was 8 per cent at 3 years. The tumour stage and grade at presentation and the proportion of screen-detected cancers were similar for both intervals. The outcome for patients before specialization (1990-1992; n = 329) was worse: hazard ratio (HR) for DFS 1.5 (95 per cent confidence interval 1.2-2.0) and HR for locoregional recurrence 2.0 (1.2-3.5). After subspecialization (1993-1996, n = 455) DFS improved from 70 to 79 per cent (P = 0.009) and the all recurrence rate fell from 22 to 12 per cent (P = 0.0004) at 3 years. The improvement in outcome was mainly in younger patients (aged less than 70 years), in whom DFS improved from 72 to 81 per cent (P = 0.02) and the all recurrence rate fell from 24 to 12 per cent (P = 0.001) at 3 years. The improvement was associated with increased axillary surgery (47 to 74 per cent; P < 0.0001), and more frequent use of tamoxifen (74 to 84 per cent; P = 0.004) and chemotherapy (10 to 27 per cent; P < 0.0001) in this age group. CONCLUSION: There was a significant improvement in outcome for patients with breast cancer after surgical subspecialization in Bedford. This may relate to the more frequent use of appropriate systemic therapy. PMID- 11044172 TI - Pattern of recurrence following radical oesophagectomy with two-field lymphadenectomy. AB - BACKGROUND: Despite increasingly radical surgery for oesophageal cancer, many patients continue to develop recurrent disease. The aim of this study was to evaluate the pattern of failure following attempted curative oesophagectomy with two-field lymphadenectomy for adenocarcinoma and squamous cell carcinoma of the middle and lower third of the oesophagus. METHODS: A total of 176 consecutive patients discharged from hospital following R0 resection between 1 April 1990 and 31 December 1999 were followed for evidence of recurrence over a mean interval of 26 months. RESULTS: Adenocarcinoma was the predominant histological subtype (n = 113) compared with squamous cell carcinoma (n = 63). Sex and age distribution were similar for both histological subtypes (M:F ratio 2.5:1, median age 64 (range 40-77) years). Overall 2- and 5-year survival rates were 54 and 31 per cent respectively. Some 85 patients (48 per cent) developed proven recurrence, of whom five are alive and 80 dead. The median time to recurrence was 11.7 (range 1. 5-67) months, with a median survival thereafter of only 2.7 (0-25.9) months. The pattern of recurrence was locoregional in 27 per cent (mediastinal 21 per cent and cervical 6 per cent) and distant in 18 per cent (liver 6 per cent, bone 6 per cent, cerebral 2 per cent, peritoneal 2 per cent, lung 1 per cent, skin 1 per cent). There was no difference in the overall pattern of dissemination or timing of recurrence for either histological subtype. Over 50 per cent of all recurrences occurred within 12 months of surgery, with local, regional and distant recurrence occurring at a median of 11.9 (range 1.8-52), 11.0 (range 5 67) and 11.0 (1.5-58) months respectively. CONCLUSION: The low incidence of cervical recurrence suggests that a more extensive 'three-field' lymphadenectomy is unlikely to improve survival rates. Better staging modalities are needed to identify patients who will have recurrence within 12 months of operation, so that they may be either entered into trials of multimodality treatment or offered non surgical palliation. British Journal of Surgery prize-winning paper, presented to the Association of Upper Gastrointestinal Surgeons of Great Britain and Ireland, London, UK, September 1999 PMID- 11044173 TI - Major intra-abdominal pathology missed at laparoscopic cholecystectomy. PMID- 11044174 TI - Changing patterns in the management of gastric volvulus over 14 years. PMID- 11044176 TI - Authors' reply PMID- 11044175 TI - Intraperitoneal polypropylene mesh repair of incisional hernia is not associated with enterocutaneous fistula. PMID- 11044177 TI - Foreword PMID- 11044178 TI - New developments in melanoma research with the discussion of practical clinical problems. PMID- 11044179 TI - Epidemiology of melanoma. AB - Melanoma incidence and morality rates are increasing in most countries throughout the world where they are being recorded. The annual incidence rates have increased in the order of 3-7% in fair-skinned populations in recent decades. The mortality rates have increased at a rate lower than for incidence. This has been attributed to educational programs designed to improve the early detection of melanoma, as the treatment of melanoma has not changed substantially in recent decades. There has been a decrease in the thickness of melanoma with an increasing proportion of thin melanomas at diagnosis. Causation of melanoma is a combination of constitutional risk factors of which skin colour is the major factor. The presence and number of common acquired and dysplastic melanocytic naevi is also a major constitutional risk factor in fair-skinned people. The only environmental risk factor that has been shown consistently is exposure to sunlight, particularly large doses of sunlight sufficient to cause sunburn in childhood that will be remembered many years later. However, recreational activity leading to sunburn in adulthood is also associated with risk. To date, no other environmental factors have been shown epidemiologically to be clearly associated with risk of melanoma. Recent epidemiological data from some studies suggesting that there is an increased risk of melanoma in sunscreen users requires further explanation. PMID- 11044180 TI - Genetics of familial and sporadic melanoma. AB - Like many other cancers, melanoma has a significant genetic basis. However, its genetic pathways may involve multiple genes with probable interactions with sun exposure. Germline mutations in p16 or CDKN2A are found in a significant percentage of relatively rare melanoma families but p16 mutations are uncommon in sporadic tumours. p16 may still be involved by other mechanisms of inactivation; however, it is clear that other melanoma genes remain to be discovered. Family, case-control, twin and sib-pair analyses as well as DNA chip technology may shed some light on genes involved in melanocytic differentiation and skin pigmentation. Recent public health campaigns have not been very successful in changing behaviour regarding tanning, and the relationship between sun exposure and melanoma is very complex. With the understanding of genetic alterations leading to this tumour, follow-up strategies and behavioural interventions may be more specifically designed to target high risk groups. PMID- 11044181 TI - Malignant melanoma: clinical variants and prognostic indicators. AB - The four main clinicopathological subsets of melanoma are described and the arguments for their retention in a research setting presented. Further additional subsets are described, including partly regressed primary melanoma, melanoma arising in a congenital naevus and multiple primary tumours. Pathological prognostic indicators are discussed and the pre-eminent significance of tumour thickness and its interaction with ulceration described. The need for accurate personal predictive profiles as distinct from those relevant to populations is discussed, as is the need for a molecular marker of metastatic potential. PMID- 11044182 TI - Surgical management of primary melanoma. AB - Melanoma precursor lesions and stage I malignant melanomas are preferentially removed by excisional surgery. Several studies have supported the concept of a more conservative excision strategy. Reduced safety margins with a maximum of 2-3 cm enable us to cover most defects by simple skin flap techniques. In critical anatomical sites and in lentigo maligna melanoma micrographic surgery has recently gained importance. The value of adjuvant surgical procedures remains controversial. Possibly, the technique of sentinel-node-biopsy provides a better approach towards a more selective use of lymphadenectomy in patients with clinically occult micrometastases. PMID- 11044183 TI - Mechanisms of metastasis. AB - The complex process of tumour cell metastasis requires a number of prerequisites. Following malignant transformation with concomitant cell cycle dysregulation, a metastatic phenotype is dependent on an altered behaviour at various cellular levels. On the one hand, this includes differences in the expression of certain adhesion molecules. Typically, a reduced expression of adhesion molecules (such as cadherins), which are responsible for cell-to-cell interactions, is observed. Simultaneously, a distinct pattern of integrin-mediated, cell-to-matrix interactions are found. These changes result in an increased migratory ability of the cells, which further depends on the controlled degradation of extracellular matrix components by proteases. In particular, the matrix metalloproteinases and the serine proteases are noteworthy in this context. Regulation of protease activity involves interactions of tumour cells with components of the extracellular matrix. Finally, interactions between tumour cells and cells of the surrounding connective tissue mediated by cytokines and growth factors are important, especially in the process of angiogenesis. Only those tumour cells which fulfil all the requirements mentioned above are able to leave the site of the primary tumour, to migrate towards the blood and lymphatic vessels, to enter the circulation and, finally, to cause distant organ metastasis. PMID- 11044184 TI - Management of regional metastases. AB - Treatment of regional melanoma metastases aims not only at local remission of the primary tumour but also at prevention or delay of distant metastases. Regional metastases can occur as satellite, in-transit and regional lymph node metastases. This is an overview of the current therapeutic modalities for regional metastatic melanoma: surgical management of relapse, satellite and in-transit metastases; therapeutic lymphadenectomy; development from elective lymph node dissection to sentinel lymph node dissection (SLND); radiotherapy and isolated limb perfusion. Instead of elective lymph node dissection the new microstaging technique by means of SLND is gaining particular importance. For this method prognostic relevance is expected and thus it is supposed to be an important staging parameter for adjuvant treatment planning. PMID- 11044185 TI - Adjuvant therapy in melanoma. AB - Melanoma is an immunologic tumour as indicated by clinical regression, long dormancy and the presence of class 1 dependent cytotoxic responses against well defined tumour peptides. The poor prognosis and relative chemoresistance of patients with regional nodal or metastatic disease highlights the urgent need for an effective adjuvant therapy. A wide variety of different agents have been assessed including high dose interferon which has been shown to improve overall survival, although results of a subsequent study have not confirmed these findings. Currently, a variety of different biotherapies and biochemotherapy regimes are being assessed in phase II and III studies and sentinel lymph node biopsy now provides an accurate method for staging so that all patients can be stratified into well-designed randomised controlled trials. PMID- 11044186 TI - Classical chemotherapy for metastatic melanoma. AB - Despite a major effort in clinical research scrutinizing various treatment regimens for patients suffering from metastatic melanoma the prognosis for these patients still remains poor. The treatment options tested have ranged from monochemotherapy, polychemotherapeutic approaches including highly toxic regimens requiring autologous bone marrow rescue, immuno-modulatory therapies, e.g. defined cytokines such as interferon-alpha and interleukin-2 as well as vaccination therapy with dendritic cells or genetically modified tumour cells, and bio-chemotherapy. Although immuno-modulatory approaches are currently regarded as the most promising, to date no improved overall survival has been achieved by any of these measures especially if tested in large multicentre trials. The focus of this review will be on classical chemotherapy with emphasis on both cutaneous and uveal melanoma. PMID- 11044187 TI - Management of malignant melanoma: new developments in immune and gene therapy. AB - Thus far, the use of classical anti-cancer treatment modalities had only rarely a beneficial impact on the prognosis of patients with metastatic melanoma. We as physicians have therefore the obligation as well as the chance to develop and test new therapeutic strategies. Our growing knowledge about the genetic basis of melanoma provides one platform to fulfil this task. Another one comes from our increasing understanding of the molecular and cellular mechanisms involved in the induction/modulation of immune responses, as well as the progress made in the field of identification of melanoma antigens, and allows for the development of a new generation of vaccines. The aim of this article is to discuss several of these new concepts towards the use of immune and gene therapy of melanoma. PMID- 11044188 TI - Visceral leprosy. PMID- 11044189 TI - Skin disease and nontuberculous atypical mycobacteria. PMID- 11044190 TI - Leprosy elimination campaign and its impending fallout. PMID- 11044191 TI - Suction blistering as a research and therapeutic tool in dermatology. PMID- 11044192 TI - Necrotizing eosinophilic folliculitis as a manifestation of the atopic diathesis. AB - BACKGROUND: Eosinophilic folliculitis (EF) is an idiopathic eruption of sterile pustules and papules involving the trunk, face, and extremities, associated in many cases with human immunodeficiency virus (HIV) infection. The classic histopathology is one of follicular-based, eosinophilic spongiosis with variable microabscess formation. We describe nine HIV-negative patients who manifested a novel form of pustular EF in the setting of atopy. MATERIALS AND METHODS: Paraffin sections of skin biopsies from ten patients, stained with hematoxylin and eosin and special preparations to evaluate for microbial pathogens, were examined. Detailed clinical histories and serologic studies were obtained. RESULTS: Among the clinical presentations in seven men, two women, and one girl (age range, 11-62 years) were ulcerative and/or nodular plaques mainly on the face and/or extremities, sometimes in an annular configuration. The clinical considerations included deep mycotic infection, ulcerative herpes, systemic vasculitis, Mucha Haberman disease, and pyoderma gangrenosum. All patients had a personal and/or family history of atopy. Co-existent medical illnesses included psoriasis, lupus erythematosus, and lymphoproliferative disease. One patient was on a calcium channel blocker, one on multiple antidepressants, and two on antihistamines, all of which are associated with immune dysregulation. All skin biopsies showed variable intra- follicular eosinophilic microabscesses, follicular necrosis, folliculocentric necrotizing eosinophilic vasculitis, marked degeneration of connective tissue fiber elements, and striking tissue eosinophilia, including flame figure formation and dermal eosinophilic abscesses. Apart from commensals, such as Pityrosporum and Demodex, microbial pathogens were not identified. CONCLUSIONS: The presentations differed from conventional EF by virtue of a strong association with atopy and by the presence of ulceration, nodule formation, follicular and dermal necrosis, and eosinophilic vasculitis. We propose the term "necrotizing eosinophilic folliculitis," and suggest that the basis of this novel form of EF is an unrepressed T-helper lymphocyte type 2 (Th2) dominant response to various epicutaneous stimuli in patients with atopy, the prototypic immune dysregulatory state associated with a Th2-dominant cytokine milieu. PMID- 11044193 TI - The significance of stress hormones (glucocorticoids, catecholamines) for eruptions and spontaneous remission phases in psoriasis. AB - BACKGROUND: In an earlier paper, it was described how acute eruptions of psoriasis may be produced in phases of immune deficiency and in the presence of bacterial antigen-releasing inflammatory foci, whereas clinical spontaneous remissions are produced in phases of immunologic activity. Therefore, it was of interest to investigate whether the stress hormones cortisol/epinephrine are involved in triggering such deficiency and activity phases. METHODS: During a series of investigations lasting up to 3 years in 95 patients, the following were determined: cortisol/epinephrine levels, polyclonal serum immunoglobulins IgM, IgG, and IgA, total serum IgE, complement C3 and C4 proteins, T cells and subpopulations, as well as streptococcal titers ASO/ADNase B, severity index (PASI) RESULTS: Phases of clinical inactivity are associated with the mechanism, "immunologic regulation," where antibacterial titers are elevated, but all other parameters are unremarkable. Eruption phases (in 32 of 95 patients) showed absolute increases in serum cortisol levels and antibacterial titers, and decreases in serum epinephrine (adrenaline) levels. Phases of spontaneous remission (in 25 of 32 patients) showed, in contrast to the eruption phases, absolute increases in serum epinephrine levels, and significant falls in serum cortisol levels and bacterial titers. CONCLUSIONS: On the basis of these results, the participation of the immune system is confirmed in the pathogenesis of psoriasis, which is subject to control by higher neurohormonal systems. Cortisol may be involved in the clinical eruption phase, and epinephrine in the remission phase. Both hormones are true antagonists and have important effects on the human immune system if produced in excess via the pituitary-adrenal axis. Infection with Streptococcus pyogenes is an additional trigger for the dermatosis. PMID- 11044195 TI - Erythema gyratum repens-like psoriasis. AB - A 28-year-old man was admitted to our department for investigation in 1992. He presented with a red, scaly, centrifugally spreading eruption, which had appeared in 1990, beginning on the neck and thorax, and later extending to the trunk and limbs. The cutaneous lesions, located mainly on the trunk and proximal upper limbs, were arranged in rings, with a slightly raised prominent scaling edge (Fig. 1a). The characteristic feature was the presence of rings or waves within already existing rings, whereas the central part was flattened, with the texture of normal skin. The concentric figurate lesions resembled a wood grain pattern (Fig. 1b). The clinical picture was strikingly similar to tinea imbricata; there was, however, no itching, and repeated mycologic studies did not disclose Trichophyton concentricum. The histology was not characteristic. The epidermis, which was slightly edematous, was covered with a heavy crust. In the dermis, a sparse inflammatory infiltrate, somewhat more pronounced in the subpapillary areas, was composed of lymphocytes with some eosinophils. Periodic acid-Schiff (PAS) and other stains for mycotic infection were negative. The general condition was not affected and laboratory studies did not show any abnormalities, except for low serum protein (5.1 g/L) and decreased gamma globulins (10.5%). Cell mediated immunity was preserved. Immunofluorescence studies (direct and indirect) were negative. In spite of repeatedly negative mycotic examinations and due to the striking similarity to tinea imbricata, we applied various antimycotic therapies (terbinafine, itraconazole), with no effect. The figurate pattern, with normal skin in between, altered from day to day, while new concentric rings appeared within the cleared skin. The migrating rate was about 2-3 cm per 2 weeks. The patient had undergone a thorough search for internal malignancy. During the follow-up period of 1992-98, cutaneous involvement slowly became almost generalized (1996), and the confluent lesions formed large plaques, but still with pronounced concentric rings. Transitional blood eosinophilia (27% in 1993 and 11% in 1996) regressed with no therapy. Since 1995, antibodies to HBs and HBc have been present with no clinical symptoms of liver disease. The blood proteins increased to 7.0 g/L, and gamma globulins to 17.2% (normal). The histology, studied repeatedly, started to display some signs of psoriasis from 1996 and, in 1998, was already consistent with the disease (Fig. 2). RE-PUVA (0.8 mg/kg acitretin and UVA 0.8 J/cm2 ) was applied for 2 weeks before the patient interrupted the therapy. In spite of this, there was further improvement and, in 1999, the patient was almost free of lesions with some abortive rings left. From time to time, single vesicles appeared within the elevated borders of the rings. The histology of such vesicles was consistent with abortive pustular psoriasis (Fig. 3). PMID- 11044194 TI - A study of cutaneous myiasis in Sri Lanka. AB - BACKGROUND AND OBJECTIVES: Cutaneous myiasis (CM) due to Diptera fly larvae shows different patterns in different regions. Many modalities of treatment have been described. The objectives of our study were to identify the species causing CM in Sri Lanka, the common sites of infestation, and the contributory factors, and to assess some treatment modalities, in particular mineral turpentine and certain herbal preparations. METHODS: All patients with CM admitted or referred to the Dermatology Unit at the General Hospital, Kalutara, over 18 months starting from July 1997, and all patients with CM from the orthopedic and surgical wards of the National Hospital of Sri Lanka in Colombo over 6 months from July 1997, were studied. Details of the history and examination were recorded on specially designed forms. Maggots extracted were identified at the Department of Parasitology, Faculty of Medicine, University of Colombo. The modalities of treatment employed in the patients were recorded. In the Department of Parasitology, a colony of Chrysomya megacephala was maintained. Homogenized leaf extracts of Azadirachta indica (neem) and Pongamia pinnata (Indian beech) and mineral turpentine (active ingredient--low aromatic white spirits) were tested for efficacy in killing C. megacephala larvae in vitro. Leaf extracts were not used directly on patients. RESULTS: There were 16 patients (10 males and five females; the sex of one patient was not recorded). The mean age was 58.5 years (range, 11-94 years). Identification of larvae revealed C. bezziana in 14 (87.5%) and C. megacephala in two (12.5%) patients. The foot was affected in 15 (93. 7%) and the scalp in one patient. The immediate predisposing factor for CM in dermatology patients was infected dermatitis. The other relevant associated factors were: diabetes mellitus, psychiatric illness, leprosy, and mental subnormality. Turpentine was a useful adjunct in the removal of maggots manually. There were no side-effects to turpentine. In the in vitro testing, turpentine was 100% effective in killing maggots. Some patients required surgical removal under anesthesia. Indian beech and neem leaf extracts were not effective against Chrysomya larvae in vitro. CONCLUSIONS: All cases of CM were due to larvae of Chrysomya species. The commonest was C. bezziana. C. megacephala larvae causing CM have been reported for the first time in Sri Lanka. The foot was the site of predilection. Dermatitis, psychiatric illness, leprosy, diabetes, and mental subnormality were some contributory factors. Topically instilled mineral turpentine, followed by manual removal of maggots, was effective in most cases. The plant extracts tested in vitro were ineffective. As C. bezziana is an obligatory parasite capable of penetrating deeply, the importance of preventive measures is emphasized. PMID- 11044196 TI - Ultrastructure of pitted keratolysis. PMID- 11044197 TI - Mucous membrane pemphigoid, thymoma, and myasthenia gravis. PMID- 11044198 TI - Genital warts at the site of healed herpes progenitalis: the isotopic response. PMID- 11044199 TI - Proteus syndrome. PMID- 11044200 TI - Self portrait of a dermatologist. PMID- 11044201 TI - Invasive hyalohyphomycosis due to Fusarium solani in a patient with acute lymphocytic leukemia. PMID- 11044202 TI - Localized idiopathic elastosis perforans serpiginosa effectively treated by the Coherent Ultrapulse 5000C aesthetic laser. PMID- 11044203 TI - Plasminogen activator inhibitor type 1 is a potential target in renal fibrogenesis. AB - Plasminogen activator inhibitor type 1 is a potential target in renal fibrogenesis. The progression of renal lesions to fibrosis involves several mechanisms, among which the inhibition of extracellular matrix (ECM) degradation appears to play an important role. Two interrelated proteolytic systems are involved in matrix degradation: the plasminogen activation system and the matrix metalloproteinase system. The plasminogen activator inhibitor type 1 (PAI-1), as the main inhibitor of plasminogen activation, regulates fibrinolysis and the plasmin-mediated matrix metalloproteinase activation. PAI-1 is also a component of the ECM, where it binds to vitronectin. PAI-1 is not expressed in the normal human kidney but is strongly induced in various forms of kidney diseases, leading to renal fibrosis and terminal renal failure. Thrombin, angiotensin II, and transforming growth factor-beta are potent in vitro and in vivo agonists in increasing PAI-1 synthesis. Several experimental and clinical studies support a role for PAI-1 in the renal fibrogenic process occurring in chronic glomerulonephritis, diabetic nephropathy, focal segmental glomerulosclerosis, and other fibrotic renal diseases. Experimental models of renal diseases in PAI-1 deficient animals are in progress, and preliminary results indicate a role for PAI-1 in renal fibrogenesis. Inhibition of PAI-1 activity or of PAI-1 synthesis by specific antibodies, peptidic antagonists, antisense oligonucleotides, or decoy oligonucleotides has been obtained in vitro, but needs to be evaluated in vivo for the prevention or the treatment of renal fibrosis. PMID- 11044204 TI - Focal segmental glomerulosclerosis associated with mitochondrial cytopathy. AB - BACKGROUND: The nonspecific lesion of focal segmental glomerulosclerosis (FSGS) can occur as a primary disease or in a variety of secondary settings. In mitochondrial cytopathies (MCs), the phenotypic expression of the disease depends on the degree of cellular dysfunction, and this correlates with the proportion of abnormal mitochondrial DNA in the cells and the dependence of tissues on oxidative metabolism. The most common renal manifestation in MCs is tubular dysfunction; little has been reported about glomerular diseases. METHODS: Cases of four adult patients with FSGS and MC are reported. Routine histology and mitochondrial DNA analysis were carried out on renal biopsies. RESULTS: Family history and clinical manifestations in the four patients with FSGS suggested a diagnosis of MC. An A3243G transition in the mitochondrial DNA tRNA(leu(UUR)) was found in lymphocytes and kidney. Glomerular lesions of FSGS were associated with unusual hyaline lesions, which appeared to represent individual myocyte necrosis in afferent arterioles and small arteries. CONCLUSION: FSGS is a renal manifestation of MCs. The renal lesion can precede other manifestations of the genetic disease by many years. The striking arteriolar lesions in these cases may have resulted in glomerular hypertension and hyperperfusion, leading to secondary epithelial cell abnormalities and, ultimately, FSGS. However, primary epithelial cell dysfunction caused by mitochondrial defects could not be ruled out on morphological grounds. MCs should be considered in cases of so-called primary FSGS, particularly if there is a familial history of diabetes, neuromuscular disorders, or deafness. PMID- 11044205 TI - Citrate therapy for polycystic kidney disease in rats. AB - BACKGROUND: Few treatments are available to slow the progression to renal failure in autosomal dominant polycystic kidney disease (PKD). In an animal model of PKD, the male heterozygous Han:SPRD rat, intake of a solution of potassium citrate plus citric acid (KCitr) from age one to three months prevented a decline in glomerular filtration rate (GFR). The present study tested whether this beneficial effect is sustained and explored handling of citrate and ammonia in normal and cystic kidneys. METHODS: Rats were provided with tap water or citrate solutions to drink, and clearance and survival studies were performed. RESULTS: The GFRs of rats with PKD that consumed KCitr from one month of age were normal at six months of age, while those of their counterparts on water were about one third of normal. Long-term KCitr treatment extended the average life span of rats with PKD from 10 to 17 months. Compared with normal rats, water-drinking rats with PKD had higher plasma [citrate], renal cortical [citrate], and fractional excretion of citrate, and lower rates of renal citrate consumption, ammonia synthesis, and ammonia excretion. Cortical PNH3 was not elevated in cystic kidneys. Intake of Na3 citrate/citric acid solution or K3 citrate solution, but not ammonium citrate/citric acid solution, prevented a decline in GFR in three month-old rats with PKD. CONCLUSIONS: Rats with PKD show abnormal renal handling of citrate and ammonia. Citrate salts that have an alkalinizing effect preserve GFR and extend survival. PMID- 11044206 TI - Autosomal dominant Alport syndrome caused by a COL4A3 splice site mutation. AB - BACKGROUND: Alport syndrome (AS) is a clinically and genetically heterogeneous renal disorder, predominantly affecting the type IV collagen alpha 3/alpha 4/alpha 5 network of the glomerular basement membrane (GBM). AS can be caused by mutations in any of the three genes encoding these type IV collagen chains. The majority of AS families (85%) are X-linked (XL-AS) involving mutations in the COL4A5 gene. Mutations in the COL4A3 and COL4A4 genes cause autosomal recessive AS (AR-AS), accounting for approximately 14% of the cases. Recently, autosomal dominant AS (AD-AS) was linked to the COL4A3/COL4A4 locus in a large family. METHODS: COL4A3 and COL4A4 cDNAs were generated by nested reverse transcription polymerase chain reaction and were analyzed by DNA sequence analysis. Denaturating high-performance liquid chromatography (DHPLC) was used for mutation and segregation analysis at the genomic DNA level. RESULTS: In the AD-AS family, a splice site mutation resulting in skipping of exon 21 of the COL4A3 gene was detected. The mutation does not alter the reading frame and is predicted to result in a COL4A3 chain with an internal deletion. CONCLUSION: As the NC domain is intact, this chain may be incorporated and distort the collagen triple helix, thereby causing the dominant effect of the mutation. The finding of a specific COL4A3 mutation in AD-AS completes the spectrum of type IV collagen mutations in all genetic forms of AS. PMID- 11044207 TI - Regulation of mesangial cell apoptosis and proliferation by intracellular Ca(2+) signals. AB - BACKGROUND: In inflammatory glomerular diseases, proliferation, as well as apoptosis of mesangial cells (MCs), has been shown histomorphologically. Both processes may regulate the cellular content of the mesangium by closely influencing each other. In the present study, we examined whether the cytoplasmic free Ca(2+) concentration [Ca(2+)](i) is involved as a key second messenger in the regulation of proliferative and apoptotic events. METHODS: Thapsigargin, an inhibitor of the endoplasmic Ca(2+)-Mg(2+)-ATPase, was used as a test substance to investigate the role of [Ca(2+)](i) in signaling MC apoptosis and growth in vitro. Apoptosis was determined by nuclear chromatin staining with Hoechst 33258, by a [3H]-thymidine-based DNA fragmentation assay or by flow cytometry detecting binding of FITC-conjugated annexin V. Proliferation was measured by [3H] thymidine incorporation into acid-precipitable material and corroborated by cell counting. RESULTS: Thapsigargin significantly induced apoptosis and inhibited proliferation dose dependently in nanomolar concentrations without evoking necrotic damage when administered not longer than 12 hours. Significant apoptosis was measurable after a six-hour treatment of MCs with thapsigargin. Determination of [Ca(2+)](i) by fura-2-dependent spectrofluorometry showed that thapsigargin was able to induce prolonged [Ca(2+)](i) rises that could be prevented by preincubation with the intracellular Ca(2+) chelator 1, 2-bis(2-aminophenoxy) ethane-N,N,N', N'-tetra-acetic acid (BAPTA) acetomethyl ester (AM). BAPTA had no influence on MC viability but reversed thapsigargin-induced apoptosis to control levels. After thapsigargin treatment (100 nmol/L, 12 hours), apoptotic MCs had a significantly higher [Ca(2+)](i) of 251 +/- 25 nmol/L (N = 41) as compared with MCs that were not or not yet apoptotic ([Ca(2+)](i) of 116 +/- 20 nmol/L, N = 26, P < 0,05). Platelet-derived growth factor (PDGF), a well-characterized growth factor for MCs, reversed the effects of thapsigargin on proliferation and apoptosis in a similar fashion as BAPTA. PDGF acutely stimulated increases of [Ca2+]i but abolished thapsigargin-dependent, but not angiotensin II- or ATP induced Ca(2+) rises when administered during a 12-hour preincubation. CONCLUSIONS: Our data suggest that a sustained increase of [Ca(2+)](i) may serve as a signal to trigger MC apoptosis. Growth factors such as PDGF can abolish apoptosis induced by elevations of [Ca(2+)](i) by altering intracellular Ca(2+) signaling. PMID- 11044208 TI - Transforming growth factor-beta 1 antisense oligodeoxynucleotides block interstitial fibrosis in unilateral ureteral obstruction. AB - BACKGROUND: Interstitial expression of transforming growth factor-beta1 (TGF beta1) is important in tubulointerstitial fibrosis, a common process in most progressive renal diseases. However, no effective therapy for progressive interstitial fibrosis is known. Recently, we developed an artificial viral envelope (AVE)-type hemagglutinating virus of Japan (HVJ) liposome-mediated retrograde ureteral gene transfer method, which allowed us to introduce the genetic material selectively into renal interstitial fibroblasts. METHOD: We introduced antisense or scrambled oligodeoxynucleotides (ODNs) for TGF-beta 1 into interstitial fibroblasts in rats with unilateral ureteral obstruction, a model of interstitial fibrosis, to block interstitial fibrosis by retrograde ureteral injection of AVE-type HVJ liposomes. RESULTS: TGF-beta 1 and type I collagen mRNA increased markedly in the interstitium of untreated obstructed kidneys, and those were not affected by scrambled ODN transfection. Northern analysis and in situ hybridization revealed that the levels of TGF-beta 1 and type I collagen mRNA were dramatically decreased in antisense ODN-transfected obstructed kidneys. Consequently, the interstitial fibrotic area of the obstructed kidneys treated with antisense ODN was significantly less than that of the obstructed kidneys untreated or treated with scrambled ODN. CONCLUSION: The introduction of TGF-beta 1 antisense ODN into interstitial fibroblasts may be a potential therapeutic maneuver for interstitial fibrosis. PMID- 11044209 TI - Axial distribution and characterization of basolateral P2Y receptors along the rat renal tubule. AB - BACKGROUND: Several groups have identified P2Y receptors in the basolateral membrane of the rat nephron. These studies have not covered all segments of the nephron and have relied solely on the relative potency of receptor agonists for classification. METHODS: We measured purine and pyrimidine-induced changes in intracellular free calcium concentration ([Ca(2+)](i)) in anatomically defined segments of the rat nephron. To complement these functional studies, we have used reverse transcription-polymerase chain reaction methodology to identify specific P2Y receptor transcripts in these segments. RESULTS: Adenosine 5'-triphosphate (ATP) mobilized [Ca(2+)](i) in all nephron segments, except for the thick ascending limb of Henle, which was poorly responsive. Adenosine (100 micromol/L) was without effect, confirming that the effect of ATP was mediated by P2 receptors. In the proximal convoluted tubule (PCT) and outer medullary collecting duct (OMCD), there was evidence for two receptor subtypes with characteristics of P2Y(1)- and either P2Y(2)- or P2Y(4)-like receptors. A novel finding in the thin limbs was the presence of a receptor with properties of both P2Y(2) and P2Y(4) receptor subtypes. To aid classification, we identified P2Y receptor mRNA in rat nephron segments. In the PCT and OMCD and thin ascending limb of Henle, we found expression of P2Y(1), P2Y(2), and P2Y(4) receptors. In the descending limb of Henle, P2Y(1) and P2Y(2) mRNA was found, but P2Y(4) was not expressed. CONCLUSION: These data suggest that extracellular ATP can influence tubular cell function in all segments of the rat nephron, through P2Y receptors via multiple (and coexpressed) P2Y receptor subtypes. PMID- 11044210 TI - Characterization of receptors for osteogenic protein-1/bone morphogenetic protein 7 (OP-1/BMP-7) in rat kidneys. AB - BACKGROUND: Osteogenic protein-1/bone morphogenetic protein-7 (OP-1/BMP-7), a member of the transforming growth factor-beta superfamily, has been shown to prevent kidney damage from ischemia/reperfusion injury in rats. The molecular events involved in OP-1 action on kidney are not yet understood. METHODS: In this study, we evaluated the biodistribution of (125)I-labeled OP-1 in rat kidneys. Adult rats received a single intravenous injection of 250 microg (125)I-labeled OP-1 per kg body wt, a dose that was effective in protecting kidneys from ischemic injury. Tissue localization, in situ hybridization, and immunostaining with a specific receptor antibody were performed to identify OP-1 cellular targets. Also, isolated plasma membranes from kidney cortex and medulla regions were analyzed to identify and characterize receptor structural components that recognize OP-1. RESULTS: At 10 and 180 minutes following injection, the relative uptake of (125)I-labeled OP-1 was consistently higher in kidney cortex than in medulla region. Upon autoradiography, kidney tissue sections revealed that OP-1 bound to the convoluted tubule epithelium, glomeruli, and collecting ducts. Moreover, in situ hybridization and immunostaining methods have shown localization of mRNA transcripts and the protein for BMP receptor type II in the cortex and medulla in similar areas as (125)I-labeled OP-1. Bulk membranes (enriched with plasma membranes) isolated from the cortex and medulla regions of kidney each bound specifically to (125)I-OP-1, and the binding of (125)I-labeled OP-1 was inhibited by unlabeled OP-1 in a dose-dependent manner. However, platelet-derived growth factor, transforming growth factor-beta, insulin-like growth factor, fibroblast growth factors, and other members of BMP family such as BMP-2 and cartilage-derived morphogenetic protein-1/growth and differentiation factor-5 (CDMP-1/GDF-5) failed to inhibit the binding of (125)I-labeled OP-1 to receptors, suggesting a high degree of specificity with which OP-1 bound to kidney receptors. Scatchard analysis of quantitative binding data indicated that the OP-1 receptors of kidney contained a single class of high-affinity binding sites for OP-1 with an association constant (Ka) of 2.26 x 109 mol/L-1 and a binding capacity of 1.01 pmol of OP-1 per mg membrane protein. When analyzed by a ligand blot technique, plasma membranes isolated from kidney cortex and medulla each showed the presence of a prominent specific band with a relative molecular mass (Mr) of 100 kD. Further analysis by Western blotting indicated that an antibody raised against BMP type II receptor effectively recognized the 100 kD OP 1 binding component of kidney plasma membranes. CONCLUSIONS: We demonstrated, to our knowledge for the first time, the presence of membrane-bound, specific, high affinity OP-1 receptors in rat kidney tissues, which are likely to mediate OP-1 actions in the kidney. The major OP-1-binding component of the kidney appears to be a long form of BMP type II receptor with a Mr of 100 kD. In vivo and in vitro evidence suggests that the cellular targets for OP-1 are convoluted tubule epithelium, glomeruli, and collecting ducts. OP-1 does not share receptor binding properties with other growth factors, including BMP-2 and CDMP-1, suggesting that its mode of action in kidney appears to be specific. PMID- 11044211 TI - Hypoxia up-regulates angiopoietin-2, a Tie-2 ligand, in mouse mesangial cells. AB - BACKGROUND: Angiopoietins are secreted factors modulating endothelial survival and morphogenesis. Our previous studies demonstrated angiopoietin-2 (Ang-2) promoter activity in vivo in maturing kidney vascular smooth muscle and mesangial cells, with Tie-2 expressed by adjacent endothelia, including glomerular capillaries. METHODS: In this study we investigated Ang-2 expression in immortalized mouse mesangial cell lines and studied the response to hypoxia. RESULTS: Using reverse transcription-polymerase chain reaction, Ang-2 and Ang-3 mRNA were detected but Ang-1 and Tie-2 transcripts were absent. As assessed by Northern and slot blotting, 8 to 24 hours hypoxia (3% O(2)) significantly increased Ang-2 mRNA levels versus normoxic (21% O(2)) cells and the rate of Ang 2 mRNA degradation was similar in both conditions, consistent with increased transcription. Hypoxia also increased immunoreactive Ang-2 in cell lysates. Hypoxic stimulation of Ang-2 mRNA was significantly reduced by inhibitors of tyrosine kinase (genistein) and protein kinase C (GF109203X), but not by a mitogen-activated protein kinase 1 inhibitor (PD98059). Furthermore, hypoxia coincidentally up-regulated levels of vascular endothelial growth factor (VEGF) mRNA in these cells. Finally, in vivo, immunoreactive Ang-2 was observed in the cores of immature glomeruli of neonatal mice, but immunostaining in this location was absent in four-week postnatal mice. CONCLUSION: This is the first demonstration that isolated mesangial cells express Ang-2 mRNA and protein and up regulate Ang-2 in response to hypoxia. We speculate that hypoxia-induced, mesangial-derived Ang-2 and VEGF may have synergistic paracrine roles in the growth of glomerular endothelia during normal development and diseases. PMID- 11044212 TI - CD44-mediated neutrophil apoptosis in the rat. AB - BACKGROUND: Apoptosis is an important mechanism by which neutrophils are removed from sites of inflammation, including the kidney. This study investigated whether ligation of the cell-surface adhesion molecule, CD44, can trigger neutrophil apoptosis. METHODS: The anti-rat CD44 antibody OX-50 was used to induce apoptosis of cultured blood neutrophils, as determined by flow cytometry using annexin V staining and by transmission electron microscopy. The functional consequences of OX-50-mediated neutrophil depletion were examined in a rat model of accelerated antiglomerular basement membrane glomerulonephritis. RESULTS: Flow cytometric analysis using the OX-50 antibody, which recognizes the common amino terminal domain of CD44, showed that rat blood neutrophils express very high levels of CD44. The addition of OX-50, but not control antibodies, rapidly induced neutrophil apoptosis in cultured rat blood leukocytes, as demonstrated by annexin V staining and by electron microscopy. Cross-linking of CD44 was essential since F(ab) fragments of the OX-50 antibody failed to induce neutrophil apoptosis. The CD44 ligand hyaluronan and an antibody to the CD44v6 isoform failed to induce neutrophil apoptosis, indicating that OX-50 antibody-mediated neutrophil apoptosis is epitope specific. This effect was specific to neutrophils since the OX-50 antibody did not induce apoptosis in other CD44-expressing cell types (lymphocytes, mesangial cells, or tubular epithelial cells). An injection of OX 50 antibody into normal rats caused a rapid and profound neutropenia, and apoptotic neutrophils could be seen in the blood by electron microscopy. Furthermore, the administration of OX-50 antibody abrogated neutrophil-dependent glomerular injury (proteinuria) on day 1 of rat antiglomerular basement membrane glomerulonephritis, whereas injury on day 10 of the disease (neutrophil independent) was largely unaffected. CONCLUSIONS: The cross-linking of specific epitopes of the CD44 molecule can rapidly induce neutrophil apoptosis in vitro and inhibit neutrophil-dependent renal injury in vivo. This finding suggests that physiological ligands of the CD44 molecule may play an important role in eliminating neutrophils from sites of inflammation, including inflammatory kidney disease. PMID- 11044213 TI - Differential expression of renal AGE-receptor genes in NOD mice: possible role in nonobese diabetic renal disease. AB - BACKGROUND: Nonobese diabetic mice (NOD) are prone to glomerular pathology, which is accelerated with the onset of diabetes. Advanced glycation end product (AGE) interactions with AGE-receptors (AGE-Rs) in kidneys can contribute to glomerular injury and diabetic nephropathy (DN). The significant elevation in kidney AGE deposits noted in prediabetic NOD mice suggested that delayed AGE turnover in this model may contribute to its propensity toward DN. METHODS: To explore whether excess tissue AGE was linked to altered AGE-R status in the kidney, mRNA/protein expression, and of several AGE-Rs [AGE-R1, AGE-R2, AGE-R3, scavenger receptor II (ScR-II), and receptor for AGE (RAGE)], was determined in renal cortex and in mesangial cells (MCs) isolated from ND-, D-NOD, and ILE mice (N = 20 per group). Ligand binding, receptor site number, and affinity were determined in MCs from the same mouse groups. RESULTS: Prediabetic NOD kidney AGE-R1 mRNA and protein level were threefold lower than that of ILE mice (P < 0.01), while AGE-R3 mRNA was enhanced by twofold (P < 0.05) and AGE-R2, RAGE, and ScR-II mRNA remained close to normal (ILE). The onset of diabetes in NOD mice, while enhancing AGE-R1 mRNA expression by approximately twofold, failed to raise it above the normal (ILE) level, despite increases in tissue, and serum AGE. The latter was associated with higher elevation in AGE-R3 (sixfold, P < 0.05), RAGE (twofold, P = NS), and ScR-II mRNA (2. 8-fold, P = NS) above control. MCs from prediabetic NOD mice showed a threefold lower level of AGE-R1 mRNA (P < 0.02 vs. ILE) and AGE-R1-protein, and AGE-binding activity (<40% of control ILE). In contrast, AGE-R3 mRNA was enhanced (twofold), while AGE-R2 showed no change. Cultured ND-NOD MCs displayed only one fourth of the AGE-binding sites/cell present on ILE MCs (1.6 x 10(6) vs. 6.6 x 10(6), P < 0.05), which after the onset of diabetes rose to the normal range (7.0 x 10(6)/cell), but failed to exceed it. CONCLUSIONS: Reduced AGE-R1 gene expression in this strain may contribute to delayed AGE removal from and early AGE deposition in kidney tissues. This may act as a trigger for those AGE-R genes involved in growth-promoting changes, leading to DN in this strain. PMID- 11044214 TI - Efficacy of galectins in the amelioration of nephrotoxic serum nephritis in Wistar Kyoto rats. AB - BACKGROUND: Galectins are characterized by specific affinity for beta-galactoside sugars, and they play a role in diverse biological processes, including cell adhesion, cell proliferation, and apoptosis. Galectin-1, -3, and -9 have been implicated in modulating the immune response. METHODS: Nephrotoxic serum nephritis, which is characterized by crescent formation and glomerular influx of CD8+ cells into glomerular capillaries, was induced in Wistar Kyoto (WKY) rats by injecting rabbit antiglomerular basement membrane serum. Following induction, the rats were treated either with phosphate-buffered saline or dexamethasone, galectin-1, galectin-3, or galectin-9 on alternate days and were sacrificed at day 14. At day 8, splenic lymphocytes were isolated and employed for terminal deoxytransferase-mediated uridine triphosphate nick end-labeling (TUNEL) assay to assess the degree of apoptosis, and the kidneys were utilized to determine the extent of influx of CD4(+) and CD8(+) cells and glomerular damage. RESULTS: Dexamethasone induced a marked apoptosis of splenic CD4(+) and CD8(+) cells, and it inhibited the production of anti-rabbit IgG and the influx of CD8+ cells and macrophages into the renal glomeruli. Crescent formation and excretion of urinary proteins were also reduced. Galectin-9 failed to induce apoptosis in the CD4(+) cells; however, it induced apoptosis in the CD8(+) cells and inhibited the infiltration of CD8(+) cells. Although galectin-1 and -3 did not induce the apoptosis in the T cells, they inhibited the accumulation of macrophages in the renal glomeruli. Like dexamethasone, the galectins also reduced the crescentic formation, proliferation of glomerular cells, and excretion of urinary proteins. CONCLUSIONS: Galectin-9 selectively induces apoptosis of the activated CD8(+) cells, while the macrophage influx into the kidney is modulated by all three galectins. This finding raises an interesting possibility for the utility of galectins in the modulation of macrophages that are involved in immune-mediated glomerular diseases. PMID- 11044215 TI - Induction of hyaluronan metabolism after mechanical injury of human peritoneal mesothelial cells in vitro. AB - BACKGROUND: Hyaluronan (HA) is an important extracellular matrix component that is involved in cell movement and tissue repair. In vertebrates, HA synthase genes (HAS 1, HAS 2, and HAS 3) that control the synthesis of HA have been identified. In this article, we investigated HA synthesis in the response of human peritoneal mesothelial cells (HPMCs) to injury. METHODS: The expression of HAS 1, HAS 2, and HAS 3 mRNA and the synthesis of [(3)H]-labeled HA were examined in an in vitro model of peritoneal mesothelial cell damage. The staining for uridine diphosphoglucose dehydrogenase, a key enzyme in the synthesis of HA, and biotinylated HA-binding protein was used to determine the cellular location of HA synthesis and its site of deposition. RESULTS: Growth-arrested human HPMCs expressed low levels of mRNA for HAS 2 and HAS 3 but not HAS 1. Following injury to the monolayer, HAS 2 was up-regulated by 6 hours, reaching maximal expression between 12 and 24 hours. In contrast, the expression of HAS 3 was down-regulated. During the same time period, synthesis of HA was increased in the injured monolayer. This synthetic activity appeared to be restricted to cells at the edge of the wound and to cells entering the wound. In a separate series of experiments, the addition of HA to the injured monolayer at a concentration range found in peritoneal fluid (50 to 3300 ng/mL) increased the migration of cells into the wound in a dose-dependent manner. CONCLUSIONS: These studies provide evidence that HA is an important component of peritoneal mesothelial cell migration. The results also suggest that in this process, there is differential regulation of HAS gene expression and that the synthesis of HA is limited to cells located at the leading edge of the wound. PMID- 11044217 TI - Components of normal serum block the focal segmental glomerulosclerosis factor activity in vitro. AB - BACKGROUND: Sera from some patients with focal segmental glomerulosclerosis (FSGS) increase glomerular albumin permeability (P(alb)) in vitro. The hypothesis that a component of normal serum can protect the glomerular permeability barrier was tested using sera from FSGS patients, normal individuals, and several mammalian and avian species. METHODS: In most experiments, isolated rat glomeruli were incubated in medium containing FSGS serum known to increase P(alb) in vitro, normal serum, or both active FSGS and normal serum. In other experiments, fractions of normal serum and serum from other vertebrate species were incubated with active FSGS serum. P(alb) was calculated from glomerular capillary expansion in response to an oncotic gradient. To enrich the blocking activity, normal pooled human plasma was subjected to various biochemical manipulations. RESULTS: Normal human serum prevented the increase in P(alb) (active FSGS sera, 0.77 +/- 0.12; active FSGS sera:normal serum, 1:1 mix, 0.06 +/- 0.30, P < 0.001). Protection diminished as the concentration of normal serum was decreased. Specific fractions of human serum, including human albumin and immunoglobulin fractions, were not protective. Blocking activity was present in 80% ammonium sulfate precipitate and certain fractions from size-exclusion chromatography of normal pooled human plasma. Normal serum from each of the vertebrate species tested also prevented the increase in P(alb). Preincubation with normal serum was protective during subsequent incubation with FSGS serum, but normal serum was not protective after preincubation with FSGS serum. CONCLUSIONS: We conclude that a factor or factors in normal serum block the permeability effect of active FSGS sera. This phenomenon may account for variability in proteinuria among patients with FSGS and may explain inconsistent proteinuria following injection of FSGS sera into experimental animals. Characterization of the protective substance(s) and the mechanism by which the increase in permeability is blocked may provide insight into the pathogenesis of FSGS. PMID- 11044216 TI - F(2)-isoprostanes mediate high glucose-induced TGF-beta synthesis and glomerular proteinuria in experimental type I diabetes. AB - BACKGROUND: The recently discovered arachidonic acid derivatives, isoprostanes, are increased in pathological conditions associated with oxidative stress, such as diabetes. No role has yet been described for isoprostanes during the development of diabetic nephropathy. Cell culture in high ambient glucose has been used as a model in elucidating cellular mechanisms underlying diabetic nephropathy. Among the growth factors involved in the effect of high glucose, transforming growth factor-beta (TGF-beta) has been described as playing a key role in the development of nephropathy. METHODS: Streptozotocin-induced diabetic rats were supplemented in their diet with the antioxidant vitamin E (1000 U/kg diet). Blood and urine samples were taken to determine renal function and isoprostane concentration, as determined by gas chromatography/mass spectrometry. Glomerular mesangial and endothelial cells were cultured in high ambient glucose to determine the synthesis of isoprostanes and the role of isoprostanes in high glucose-induced synthesis of TGF-beta. RESULTS: Streptozotocin-induced diabetic rats had marked increases in plasma levels and urinary excretion rates of F(2) isoprostanes. Dietary supplementation with vitamin E normalized (plasma) and reduced (urine) isoprostane levels and, surprisingly, improved proteinuria and blood urea nitrogen (BUN) levels. High ambient glucose increased F(2)-isoprostane synthesis in glomerular endothelial and mesangial cells in culture. Incubation of glomerular cells with F(2)-isoprostanes stimulated the production of TGF-beta. CONCLUSIONS: Increased F(2)-isoprostane synthesis during diabetes appears to be responsible in part for the increase in renal TGF-beta, a well-known mediator of diabetic nephropathy. PMID- 11044218 TI - Association of decreased calcium-sensing receptor expression with proliferation of parathyroid cells in secondary hyperparathyroidism. AB - BACKGROUND: The down-regulation of both calcium-sensing receptor (CaSR) and vitamin D receptor (VDR) in parathyroid (PT) glands of secondary hyperparathyroidism (HPT) caused by chronic renal failure has been associated with PT hormone hypersecretion as well as PT hypergrowth. To clarify the predominance of decreased expression of CaSR and VDR in the high proliferative activity of PT glands, we examined the relationship between the expression of both receptors and proliferative activity in human PT glands. METHODS: Serial sections of 56 PT glands, including 52 glands from secondary HPT and 4 normal PT glands resected together with thyroid carcinoma, were examined immunohistochemically with specific antibodies against CaSR, VDR, and Ki67. The Ki67-positive cell number was counted and expressed as the Ki67 score. The CaSR and VDR expressions were semiquantitatively analyzed. RESULTS: The expressions of both CaSR and VDR were markedly decreased in PT glands of secondary HPT, while the Ki67 score was significantly higher than it was in normal controls. When hyperplastic glands were classified into two subgroups, with [N(+)] or without [N(-)] nodular formation, CaSR expression was significantly decreased in N(+), while VDR expression was not different. Multiple regression analyses revealed that the decreased expression of CaSR could contribute significantly to the high proliferative activity, even if VDR expression was taken into account. CONCLUSION: The decrease in CaSR expression is associated with the high proliferative activity of PT glands in secondary HPT, independently of the decreased VDR expression. These findings provide a new insight into the pathogenesis of PT hyperplasia, which is refractory to vitamin D therapy in patients with severe secondary HPT. PMID- 11044219 TI - Unilateral ureteral obstruction in neonatal rats leads to renal insufficiency in adulthood. AB - BACKGROUND: Although unilateral ureteropelvic junction obstruction is the most common cause of congenital obstructive nephropathy in infants and children, management remains controversial, and follow-up after pyeloplasty is generally limited to the pediatric ages. We have developed a model of temporary unilateral ureteral obstruction (UUO) in the neonatal rat: One month following the relief of five-day UUO, the glomerular filtration rate (GFR) of the postobstructed kidney was normal despite a 40% reduction in the number of glomeruli and residual vascular, glomerular, tubular, and interstitial injury. METHODS: To determine whether hyperfiltration and residual injury of remaining nephrons leads to progression of renal insufficiency in later life, 31 rats were sham operated or subjected to left UUO at one day of age, with relief of UUO five days later, and were studied at one year of age. GFR was measured by inulin clearance, and the number of glomeruli, tubular atrophy, glomerular sclerosis, and interstitial fibrosis were measured by histomorphometry in sham, obstructed (UUO), and intact opposite kidneys. Intrarenal macrophages and alpha-smooth muscle actin were identified by immunohistochemistry. RESULTS: Despite relief of UUO, ultimate growth of the postobstructed kidney was impaired. The number of glomeruli was reduced by 40%, and GFR was decreased by 80%. However, despite significant compensatory growth of the opposite kidney, there was no compensatory increase in GFR, and proteinuria was increased. Moreover, glomerular sclerosis, tubular atrophy, macrophage infiltration, and interstitial fibrosis were significantly increased not only in the postobstructed kidney, but also in the opposite kidney. CONCLUSIONS: Although GFR is initially maintained following relief of five-day UUO in the neonatal rat, there is eventual profound loss of function of the postobstructed and opposite kidneys because of progressive tubulointerstitial and glomerular damage. These findings suggest that despite normal postoperative GFR in infancy, children undergoing pyeloplasty for ureteropelvic junction obstruction should be followed into adulthood. Elucidation of the cellular response to temporary UUO may lead to improved methods to assess renal growth, injury, and functional reserve in patients with congenital obstructive nephropathy. PMID- 11044220 TI - Evidence for Rho protein regulation of renal tubular epithelial cell function. AB - BACKGROUND: Rho proteins are small guanine 5'-triphosphate (GTP)-binding proteins felt to be important regulators of several aspects of cell function, including the organization of the actin cytoskeleton. The effects of Rho proteins on the regulation of renal tubular epithelial cell function are not known. METHODS: Selected bacterial toxins that inhibit Rho protein function were used to examine the effect of Rho in cultured renal tubular epithelial cells. RESULTS: Clostridium difficile toxin A significantly and dose dependently inhibited LLC PK(1) cell (3)H-thymidine uptake and healing of small wounds made in confluent monolayers, and it induced apoptosis. A second Clostridium difficile toxin (toxin B) that acted via a different receptor also impaired LLC-PK(1) thymidine uptake and wound healing, and it induced apoptosis. A third bacterial toxin, C3 toxin from Clostridium botulinum, also impaired LLC-PK(1) thymidine uptake and stimulated apoptosis in LLC-PK(1) cells. Since Rho inhibition disrupted organization of the actin cytoskeleton, we examined the effects of another agent that disrupted the actin cytoskeleton (cytochalasin D) and found significant dose dependent effects that impaired LLC-PK1 thymidine uptake and wound healing and that induced apoptosis. The effects of toxin A and cytochalasin D to induce apoptosis were not associated with significant changes in expression of Bcl-2, BAD, or BAK proteins and were significantly attenuated by a pancaspase inhibitor. CONCLUSIONS: Our results suggest that Rho proteins are important endogenous regulators of several aspects of renal tubular epithelial cell function, including proliferation, migration, and apoptosis. Further studies are needed to clarify the cellular mechanisms of Rho regulation of renal epithelial cell function. PMID- 11044221 TI - Induction of apoptosis during development of hypertensive nephrosclerosis. AB - BACKGROUND: As the biology of programmed cell death, or apoptosis, is clarified, a role for this process in the pathophysiology of organ dysfunction and fibrosis has been hypothesized. Hypertensive nephrosclerosis represents an important cause of end-stage renal disease. One model of the progressive, noninflammatory, sclerotic renal lesion of hypertension is the Dahl/Rapp salt-sensitive rat, which was examined in this study. METHODS: Male, Dahl/Rapp salt-sensitive (SS) and Sprague-Dawley rats were placed on either 0.3 or 8.0% NaCl diets for three weeks. Blood pressure was determined, and the kidneys were harvested for histochemical analysis and to obtain total RNA for RNase protection assays and total protein for Western blotting. RESULTS: An increase in apoptosis in the glomerular and tubular compartments was observed only in kidneys of SS rats on the high-salt diet. These findings occurred at a time when renal function was markedly impaired and irreversible changes in renal morphology developed. Temporally associated with this increase in apoptosis was augmented expression of pro-apoptotic molecules that included Fas, Bax, and Bcl-XS. CONCLUSIONS: The inappropriate shift in expression of proteins that facilitate apoptosis in the nephron, along with ongoing cell death that manifested at a time when renal function was deteriorating, supported an important role for this process in development of hypertensive nephrosclerosis. PMID- 11044222 TI - Salutary role for angiotensin in partial urinary tract obstruction. AB - BACKGROUND: In the neonatal period, angiotensin II (Ang II) is up-regulated and induces a timely development of the renal pelvis and ureteral peristalsis, thereby protecting the kidney from hydronephrosis. We tested the possibility that in adulthood, Ang II may act salutarily on the kidney structure during partial urinary tract obstruction by inducing adaptive changes in the peristaltic machinery. METHODS: Adult male Sprague-Dawley rats were subjected to partial unilateral ureteral obstruction (UUO) and divided into two groups, that is, those treated with (group L, N = 21) and those without (group C, N = 21) an angiotensin type 1 (AT1) receptor antagonist (losartan). Control animals were sham operated (N = 10). Rats were sacrificed either at day 7 or day 14. RESULTS: The degree of hydronephrosis determined morphometrically was significantly more severe in group L than group C at both day 7 and day 14, indicating that Ang II inhibition accentuated hydronephrosis. The measurement of upstream pressure within the partially ligated ureter in vivo revealed that losartan significantly attenuates the frequency of ureteral peristaltic activities. In in vitro studies using ureteral strips harvested from normal adult Sprague-Dawley rats (N = 10), Ang II (10(-8) mol/L) was shown to augment contraction, which was completely inhibited by losartan (10(-6) mol/L). CONCLUSIONS: Ang II has a salutary effect of protecting kidneys from hydronephrosis during partial ureteral obstruction through its ability to augment ureteral peristalsis. PMID- 11044224 TI - High glucose accelerates the life cycle of the in vivo exposed mesothelium. AB - BACKGROUND: Mouse mesothelium exposed in vivo for 30 days to high glucose solutions develop morphological changes that characterize a population of cells near the end of their life span. METHODS: The present study was designed to explore, in mesothelial cell imprints, whether these changes could derive from an early acceleration of the cell population life cycle in mice exposed for periods of up to 30 days to a 4.25% glucose fluid (236 mmol/L/L) prepared in Hank's balanced salt solution (HBSS). Three critical points of the cell's life cycle were evaluated: the G1 checkpoint [proliferating cell nuclear antigen (PCNA) expression], DNA synthesis ((3)H-thymidine incorporation), and the prevalence of mitosis. RESULTS: Cell populations exposed to a high glucose concentration showed an initial acceleration of their life cycle, as sustained by a peak of mitosis at two hours, an early increase of DNA incorporation sustained during the first 24 hours, as well as a top level of PCNA expression after three to four hours. These significantly higher values, compared with the control animals treated with HBSS, collapsed after 24 hours and were nil after 30 days of exposure. CONCLUSIONS: Exposure to a high glucose concentration induced an early and short-lived acceleration of the mesothelial cell cycle, and with a longer exposure this was followed by a depletion of the growth capabilities of the exposed monolayer. PMID- 11044223 TI - Endogenous hepatocyte growth factor ameliorates chronic renal injury by activating matrix degradation pathways. AB - BACKGROUND: Hepatocyte growth factor (HGF) has been shown to promote tubule repair and renal regeneration following acute injury; however, whether HGF also modulates the development and progression of chronic renal diseases that are characterized by progressive tissue fibrosis is uncertain. To examine this question, this study investigated the functional consequence of blocking endogenous HGF signaling in vivo in a model of chronic renal disease. The effects of HGF on the processes of matrix synthesis and degradation in cultured renal epithelial cells were also examined. METHODS: The level of activity of the HGF/c met axis was examined in rats following 5/6 nephrectomy at multiple time points. To determine the effects of HGF in modulating chronic renal injury, HGF action was blocked in remnant kidney rats using an anti-HGF antibody. The effects of HGF on extracellular matrix (ECM) synthesis and degradation were examined in renal epithelial cells by (35)S-methionine labeling, Western blotting, and zymographic analysis. RESULTS: An increase in renal and systemic production of HGF coupled with an increase in renal c-met was observed in rats with remnant kidneys. When HGF action was blocked by the administration of an anti-HGF antibody, rats experienced a rapid decrease in glomerular filtration rate and increased renal fibrosis. Kidney sections from the antibody-treated rats displayed a marked increase in ECM accumulation and in alpha-smooth muscle actin-positive cells in both the interstitium and tubular epithelium. In vitro studies revealed that HGF reduced net ECM accumulation by human proximal tubule cells (HKC), and this effect was abolished by incubating cells with an anti-HGF antibody. HGF did not alter the ECM synthetic rate in HKC cells. Rather, it markedly increased collagenase such as matrix metalloproteinase-9 (MMP-9) protein expression, as evidenced by Western blotting and zymographic analysis. HGF also decreased the expression of tissue inhibitors of matrix metalloproteinase-1 (TIMP-1) and TIMP 2, the endogenous inhibitors of MMPs. CONCLUSION: These results suggest that HGF is a potent antifibrogenic factor both in vitro and in vivo. Endogenous activation of HGF tends to preserve kidney structure and function in rats with chronic renal disease by activating matrix degradation pathways. PMID- 11044225 TI - Role of macula densa nitric oxide and cGMP in the regulation of tubuloglomerular feedback. AB - BACKGROUND: Previous studies have suggested that nitric oxide (NO) produced within cells of the macula densa (MD) modulates tubuloglomerular feedback (TGF). We tested the hypothesis that NO produced in the MD acts locally as an autacoid to activate soluble guanylate cyclase and cGMP-dependent protein kinase in the MD itself. METHODS: Rabbit afferent arterioles (Af-Arts) and attached MD were simultaneously microperfused in vitro. The TGF response was determined by measuring the Af-Art diameter before and after increasing NaCl in the MD perfusate (from 17 mmol/L of Na and 2 of Cl to 65 mmol/L of Na and 50 of Cl). TGF was studied before (control TGF) and after inhibiting components of the NO-cGMP dependent cascade in the tubular or vascular compartment. RESULTS: Increasing NaCl concentration in the MD perfusate decreased the Af-Art diameter by 3.2 +/- 0.5 microm (from 18.5 +/- 1.3 to 15.4 +/- 1.3 microm, P < 0.001). Adding a soluble guanylate cyclase inhibitor (LY83583) to the MD increased TGF response to 6.3 +/- 1.1 microm (P < 0.031 vs. control TGF). Similarly, when cGMP-dependent protein kinase was inhibited with KT5823, TGF was augmented from 2.6 +/- 0.3 to 4.0 +/- 0.7 microm (P < 0.023). An analogue of cGMP in the MD reversed the TGF potentiating effect of both 7-nitroindazole (7NI; an nNOS inhibitor) and LY83583. Inhibition of MD guanylate cyclase did not alter the effect of acetylcholine (a NO-cGMP-dependent vasodilator) on the Af-Art. Perfusing the Af-Art with the guanylate cyclase inhibitor did not potentiate TGF, suggesting that the effect of NO occurred at the MD via a cGMP-dependent mechanism. To determine whether the effect of NO in the MD was entirely mediated by cGMP, TGF was studied after giving (1) LY83583 or (2) LY83583 plus 7NI. Adding the nNOS inhibitor to the MD did not potentiate the TGF response further. CONCLUSIONS: We concluded the following: (1) NO produced by the MD inhibits TGF via stimulation of soluble guanylate cyclase, generating cGMP and activating cGMP-dependent protein kinase; (2) NO acts on the MD itself rather than by diffusing to the Af-Art; and (3) most, if not all, of the effect of NO in the MD is due to a cGMP-dependent mechanism rather than to other NO mediators. PMID- 11044226 TI - Hyaluronan content in the kidney in different states of body hydration. AB - BACKGROUND: Growing evidence suggests that the interstitial hyaluronan (HA) content is a determinant of the fluid exchange barrier in tissues through its high resistance to water flow. This study addressed the possible involvement of renal papillary HA in water balance regulation. METHODS: In anesthetized rats during different states of renal water handling (euvolemia, water diuresis, antidiuresis), in desert rodents, and in Brattleboro rats (diabetes insipidus) with a hereditary difference in water handling, regional renal HA and water contents were measured. RESULTS: The intrarenal HA distribution is heterogeneous, with 100 times larger amounts in the papilla than in the cortex. Compared with control rats, two hours of water diuresis increased the papillary HA content by 48% and that in the outer medulla by 52%, leaving the cortex unaffected. After 24 hours of water deprivation, papillary HA was decreased by 17%, while outer medullary HA remained unchanged. In gerbils, papillary and outer medullary HA contents were only 25 and 13%, respectively, of those in normal rats, while the cortical content was similar. In Brattleboro rats, the outer medullary HA content was significantly higher (285%) than in the normal rat, while the papillary content was similar. Generally, papillary HA was positively correlated to water content but was inversely related to urine osmolality. CONCLUSIONS: The amount of renal papillary HA changes in response to water balance of the organism. When excess water needs to be excreted, increased papillary interstitial HA could antagonize water reabsorption. The opposite occurs during water conservation. HA may play a role in renal water handling by affecting physicochemical characteristics of the papillary interstitial matrix and influencing the interstitial hydrostatic pressure, thereby determining interstitial water diffusion. PMID- 11044227 TI - Autocrine effects of nitric oxide on HCO(3)(-) transport by rat thick ascending limb. AB - BACKGROUND: In vivo and in vitro studies have shown that nitric oxide (NO) is an important modulator of transport processes along the nephron. The thick ascending limb (TAL) plays a significant role in the urine-concentrating mechanism and in the maintenance of acid/base balance. METHODS: TALs from male Sprague-Dawley rats were isolated and perfused, and net bicarbonate flux (J(HCO3)(-) was determined. RESULTS: In perfused TALs, 0.5 mmol/L L-arginine (L-Arg), the substrate for NO synthase, significantly lowered J(HCO3)(-) from 35.4 +/- 4.6 to 23.2 +/- 2.9 pmol. mm(-1). min(-1), a decrease of 36.9 +/- 11.6% (P < 0.025). D-Arg (0.5 mmol/L) had no effect on J(HCO3)(-) (N = 7). In the presence of 5 mmol/L L-NAME, an NO synthase (NOS) inhibitor, the addition of L-Arg did not affect TAL J(HCO3)( ) (43.4 +/- 4.4 vs. 44.6 +/- 5.0 pmol. mm(-1). min(-1)). L-NAME alone (5 mmol/L) did not affect TAL J(HCO3)(-). After removing L-Arg from the bath, J(HCO3)(-) increased from 26.2 +/- 3.9 to 34.8 +/- 3.2 pmol. mm(-1). min(-1) (P < 0.01), indicating no cytotoxicity of NO. We next investigated the effect of cGMP analogues on TAL J(HCO3)(-). 8-Br-cGMP (50 micromol/L) and db-cGMP (50 micromol/L) significantly decreased J(HCO3)(-) by 26.3 +/- 9.1% and 35.1 +/- 11.6%, respectively. In the presence of cGMP (50 micromol/L), the addition of L Arg had no effect on J(HCO3)(-). In the presence of KT-5823 (2 mircromol/L), a protein kinase G inhibitor, the addition of L-Arg did not change TAL J(HCO3)(-) (N = 5). CONCLUSIONS: We conclude that (1) endogenously produced NO inhibits TAL J(HCO3)(-) in an autocrine manner, (2) cGMP mediates all the effects of NO, and (3) this effect is mediated by protein kinase G activation. PMID- 11044228 TI - Renal expression of constitutive NOS and DDAH: separate effects of salt intake and angiotensin. AB - BACKGROUND: Nitric oxide (NO) is generated from NO synthase (NOS) isoforms. These enzymes can be inhibited by asymmetric dimethylarginine, which is inactivated by N(G)-N(G)-dimethylarginine dimethylaminohydrolase (DDAH). The neuroneal (nNOS) type I and endothelial (eNOS) type III constitutive NOS isoforms are expressed predominantly in the macula densa and microvascular endothelium of the renal cortex, respectively. DDAH is expressed at sites of NOS expression. Since NO may coordinate the renal responses to angiotensin II (Ang II) and changes in salt intake, we tested the hypothesis that salt intake regulates the expression of nNOS, eNOS and DDAH by Ang II acting on type 1 (AT(1)) receptors. METHODS: Groups (N = 6) of rats were adapted to low-salt (LS) or high-salt (HS) intakes for 10 days. Other groups of LS and HS rats received the AT(1) receptor antagonist losartan for six days (to test the effects of salt independent of AT(1) receptors). A further group of HS rats received an infusion of Ang II for six days (to test the effect of Ang II independent of salt intake). RESULTS: Compared with HS rats, there was a significant (P < 0.05) increase in LS rats of nNOS protein in kidney and immunohistochemical expression in the macula densa, and of eNOS protein expression and immunohistochemical expression in the microvascular endothelium, and of DDAH protein expression. Losartan prevented these effects of salt on the expression of eNOS or DDAH, both of which were also increased by Ang II infusions in HS rats. In contrast, losartan did not prevent the effects of salt on nNOS expression, which was unresponsive to Ang II infusion. The generation of NO(2)(-) released by slices of renal cortex, in the presence of saturating concentrations of L-arginine, was increased by LS, compared to HS, independent of losartan and by Ang II during HS. CONCLUSION: The expressions of eNOS in cortical microvascular endothelium and DDAH in kidney are enhanced by Ang II acting on AT(1) receptors. The expression of nNOS in the macula densa is enhanced by salt restriction independent of Ang II or AT(1) receptors. PMID- 11044229 TI - ACE inhibition or angiotensin receptor blockade: impact on potassium in renal failure. VAL-K Study Group. AB - BACKGROUND: Inhibition of the renin-angiotensin system is known to raise serum potassium [K(+)] levels in patients with renal insufficiency or diabetes. No study has evaluated the comparative effects of an angiotensin-converting enzyme (ACE) inhibitor versus an angiotensin receptor blocker (ARB) on the changes in serum [K(+)] in people with renal insufficiency. METHODS: The study was a multicenter, randomized, double crossover design, with each period lasting one month. A total of 35 people (21 males and 14 females, 19 African Americans and 16 Caucasian) participated, with the mean age being 56 +/- 2 years. Mean baseline serum [K(+)] was 4.4 +/- 0.1 mEq/L. The glomerular filtration rate (GFR) was 65 +/- 5 mL/min/1.73 m(2), and blood pressure was 150 +/- 2/88 +/- 1 mm Hg. The main outcome measure was the difference from baseline in the level of serum [K+], plasma aldosterone, and GFR following the initial and crossover periods. RESULTS: For the total group, serum [K(+)] changes were not significantly different between the lisinopril or valsartan treatments. The subgroup with GFR values of < or = 60 mL/min/1.73 m(2) who received lisinopril demonstrated significant increases in serum [K(+)] of 0.28 mEq/L above the mean baseline of 4.6 mEq/L (P = 0.04). This increase in serum [K(+)] was also accompanied by a decrease in plasma aldosterone (P = 0.003). Relative to the total group, the change in serum [K(+)] from baseline to post-treatment in the lisinopril group was higher among those with GFR values of < or = 60 mL/min/1.73 m(2). The lower GFR group taking valsartan, however, demonstrated a smaller rise in serum [K(+)], 0.12 mEq/L above baseline (P = 0.1), a 43% lower value when compared with the change in those who received lisinopril. This blunted rise in [K(+)] in people taking valsartan was not associated with a significant decrease in plasma aldosterone (P = 0.14). CONCLUSIONS: In the presence of renal insufficiency, the ARB valsartan did not raise serum [K(+)] to the same degree as the ACE inhibitor lisinopril. This differential effect on serum [K(+)] is related to a relatively smaller reduction in plasma aldosterone by the ARB and is not related to changes in GFR. This study provides evidence that increases in serum [K(+)] are less likely with ARB therapy compared with ACE inhibitor therapy in people with renal insufficiency. PMID- 11044230 TI - Pretreatment blood pressure reliably predicts progression of chronic nephropathies. GISEN Group. AB - BACKGROUND: Random, nontimed blood pressure (BP) measurements in the outpatient clinic may fail to provide reliable information on actual daily BP control in renal patients on chronic antihypertensive therapy. METHODS: In a cohort of 163 patients with proteinuric chronic nephropathies followed prospectively with repeated BP and glomerular filtration rate (GFR) measurements, we compared baseline and follow-up pretreatment, morning ("trough," measured by standard procedures, and "0 minutes," measured by an automatic device) and post-treatment (120 minutes) measurements, with BP monitored up to 600 minutes after treatment administration. We then evaluated which BP value most reliably predicted GFR decline (delta GFR) and progression to end-stage renal failure (ESRF) over a median (interquartile range) follow-up of 20 (9 to 25) months. RESULTS: GFR decline was more reliably predicted by systolic as compared with diastolic BP and by pretreatment as compared to post-treatment BP, regardless of the timing and method of measurement, respectively. In particular, at the 120-minute baseline and follow-up measurements, systolic BP had no predictive value in patients with less severe renal insufficiency and baseline diastolic BP, regardless of the level of renal dysfunction. The BP predictive value was remarkably higher in ramipril than in conventionally treated patients. All follow-up-but no baseline measurements reliably predicted the risk of ESRF in the entire study group. CONCLUSIONS: In patients with progressive chronic nephropathies, systolic BP and pretreatment morning BP measurements are the most reliable predictors of disease outcome and may serve to guide antihypertensive therapy in routine clinical activities and in prospective controlled trials, particularly in patients on angiotensin-converting enzyme inhibitor therapy. Reliability and relevance of single measurements taken at different times after treatment administration are questionable. PMID- 11044232 TI - Influence of obesity on the appearance of proteinuria and renal insufficiency after unilateral nephrectomy. AB - BACKGROUND: Some patients develop proteinuria and progressive renal failure after unilateral nephrectomy, although the majority of patients maintain normal renal function. Reasons to explain this different evolution are not known. METHODS: A cross-sectional study was performed in 73 patients who had undergone unilateral nephrectomy 13.6 +/- 8.6 years before. Patients with morphologic abnormalities in the remaining kidney, systemic disorders, or abnormal renal function at the time of nephrectomy were excluded. All of the 73 included patients showed normal renal function and negative proteinuria at nephrectomy. The patient's medical records were reviewed, and clinical and analytical data throughout follow-up were obtained. RESULTS: Fifty-three out of the 73 patients (group I) showed a normal renal function and negative proteinuria at the cross-sectional study. The remaining 20 patients (group II) showed proteinuria (3.4 +/- 3.1 g/day). The time elapsed between nephrectomy and proteinuria appearance was 10.1 +/- 6.1 years. Thirteen patients of group II had developed renal insufficiency (serum creatinine at the cross-sectional study of 3.9 + 3.2 mg/dL) in addition to proteinuria. The time elapsed between proteinuria appearance and the onset of renal insufficiency was 4.1 +/- 4.3 years. Renal insufficiency showed a slowly progressive course in most of these patients. There were no significant differences between group I and group II patients in age, gender, renal function, or blood pressure at the time of nephrectomy. In contrast, group II patients showed a body mass index (BMI) that was significantly higher than group I at nephrectomy (31.6 +/- 5.6 vs. 24.3 +/- 3.7 kg/m(2), P < 0.001), at cross-sectional study (33.3 +/- 6.6 vs. 25.1 +/- 3.5 kg/m(2), P < 0.001), and throughout follow-up. Among the 14 obese (BMI > 30 kg/m(2)) patients at the time of nephrectomy, 13 (92%) developed proteinuria/renal insufficiency. In contrast, among the 59 patients with BMI < 30 kg/m(2), only 7 (12%) developed these complications (P < 0.001). Kaplan-Meier estimated probability of negative proteinuria and normal renal function 10 years after nephrectomy was 40 and 70%, respectively, in obese patients at nephrectomy. At 20 years after nephrectomy, these percentages were 8 and 35%, respectively. In contrast, in nonobese patients, the probability of negative proteinuria and normal renal function was 93 and 98%, respectively, at 10 years (P < 0.001) and 77 and 91%, respectively, at 20 years (P < 0.001). Multiple logistic regression analysis showed that the risk of developing renal disease was only statistically correlated with BMI at the time of unilateral nephrectomy (odds ratio 1.34, 1.03 to 1.76 CI). CONCLUSIONS: Obese patients are at risk for developing proteinuria and chronic renal failure after unilateral nephrectomy. Regular and long-term follow-up are recommended in these patients. PMID- 11044231 TI - Etiology of nephrocalcinosis in preterm neonates: association of nutritional intake and urinary parameters. AB - BACKGROUND: Nephrocalcinosis (NC) in preterm neonates has been described frequently, and small-scale studies suggest an unfavorable effect on renal function. The etiologic factors have not yet been fully clarified. We performed a prospective observational study to identify factors that influence the development of NC. METHODS: The study population consisted of 215 preterm neonates with a gestational age <32 weeks. Clinical characteristics and intake in the first four weeks of calcium, phosphorus, vitamin D, protein, and ascorbic acid were noted. Serum calcium, phosphate, vitamin D, magnesium, uric acid, creatinine, urea and urinary calcium, phosphate, oxalate, citrate, magnesium, uric acid, and creatinine were assessed at four weeks of age and at term. Renal ultrasonography (US) was performed at four weeks and at term. At term was defined as a postconceptional age of 38 to 42 weeks. RESULTS: NC was diagnosed by means of US in 33% at four weeks and in 41% at term. Patients with NC at four weeks had a significantly higher mean intake of calcium (P < 0.05), phosphorus (P < 0.05), and ascorbic acid (P < 0.01) than patients without NC. They had a higher mean serum calcium (2.55 vs. 2.46 mmol/L, P < 0.01) and a higher mean urinary calcium/creatinine ratio (2.6 vs. 2.1 mmol/mmol, P < 0.05). Patients with NC at term had a lower birth weight (1142 vs. 1260 g, P < 0.05) and a lower gestational age (28.8 vs. 29.4 weeks, P < 0.05), were treated significantly longer with furosemide, dexamethasone, theophylline, and thiazides, developed chronic lung disease more frequently (40 vs. 16%, P < 0.001), and had a higher mean urinary calcium/creatinine ratio (2.7 vs. 2.3 mmol/mmol, P < 0.05) and a lower mean urinary citrate/calcium ratio (1.1 vs. 1.7 mmol/mmol, P = 0.005). CONCLUSIONS: NC develops as a result of an imbalance between stone-inhibiting and stone-promoting factors. A high intake of calcium, phosphorus, and ascorbic acid, a low urinary citrate/calcium ratio, a high urinary calcium/creatinine ratio, immaturity, and medication to prevent or treat chronic lung disease with hypercalciuric side effects appear to contribute to the high incidence of NC in preterm neonates. PMID- 11044233 TI - Cross-sectional and longitudinal predictors of serum albumin in hemodialysis patients. AB - BACKGROUND: Lower serum albumin concentrations predict increased mortality in hemodialysis (HD) patients. Many demographic, comorbidity, and modifiable treatment-related factors that predict HD patient outcomes may be associated with serum albumin. METHODS: Cross-sectional predictors of baseline albumin on December 31, 1993 were sought (N = 3981). Additional effects of the same baseline predictors on subsequent trends in albumin over one year were examined in a nested subsample of patients (N = 2245). Wave-1 of the United States Renal Data System Dialysis Morbidity and Mortality special study provided the data. RESULTS: Significant associations (P < 0.05) are summarized as older age, female gender, peripheral vascular disease, chronic obstructive pulmonary disease, and cancer predicted a lower baseline albumin and negatively influenced subsequent albumin trends. Baseline albumin was higher for blacks (vs. whites), lower for smoking and diabetes, and lower during the first year of HD treatment (<3 months and 3 to 12 months, vs.> 1 year). Trend analysis showed more positive albumin slopes for patients in their first year on HD and more negative slopes for Native Americans (vs. whites). Baseline albumin was correlated with the type of vascular access being used [arteriovenous (AV) fistulas > AV grafts > permanent catheters > temporary catheters]. Trend analysis predicted more negative albumin slopes for AV grafts and permanent catheters (vs. AV fistula access). Baseline albumin correlated inversely with bicarbonate and directly with hematocrit. Dialysis with unmodified cellulose membranes, without reuse, predicted lower baseline albumin than the other membrane-reuse categories. CONCLUSIONS: Several exposures, which may be modifiable, were associated with serum albumin. PMID- 11044234 TI - Glomerular permselectivity in early stages of overt diabetic nephropathy. AB - BACKGROUND: Impairment of glomerular size selectivity has been demonstrated by the dextran-sieving technique in nephropathic diabetics with heavy, but not mild proteinuria. The purpose of the present study was to determine whether such a barrier defect could be demonstrated with mild proteinuria by substituting Ficoll 70, a polysucrose, for dextran as a probe of the filtration barrier. METHODS: Differential solute clearances were performed in 12 individuals with early diabetic nephropathy on two occasions: after 60 days of treatment with losartan 50 mg daily or a placebo. An uncharged preparation of nonreabsorbable Ficoll 70 was infused along with inulin. Fractional clearance (theta) of Ficoll of discrete size was determined after separating molecules in urine and plasma in narrow 2 A fractions over a 20 to 60 A radius interval by size exclusion high-performance liquid chromatography (HPLC). A hydrodynamic theory of hindered ficoll transport through water-filled pores was used to characterize the pore size distribution of the glomerular barrier. RESULTS: The theta for Ficoll molecules with radii> 50 A was selectively enhanced in placebo-treated diabetic nephropathy versus corresponding theta in healthy control subjects (N = 12). Computations revealed a lower distribution of glomerular pores that was unaltered in nephropathic diabetics. However, an upper distribution of large, shunt-like pores was more prominent, exceeding healthy controls by one order of magnitude in diabetic nephropathy (P = 0.01). A trend to lower theta for Ficoll in the 56 to 60 A radius range during losartan therapy is computed to have lowered the fraction of shunted filtrate by 26 to 44%, depending on whether glomerular pressure declined. The corresponding reduction in theta for endogenous albumin, IgG, and IgG4 was by 19 to 23% (P < 0.05). CONCLUSION: Our findings suggest that shunt-like defects, partially reversible by angiotensin II blockade, are present early in the course of diabetic nephropathy. We estimate that such defects can account for immunoglobulinuria in this disorder. Additional impairment of either charge- or shape-selectivity must be invoked to explain the observed level of albuminuria, however. PMID- 11044235 TI - Multicompartment urea kinetics in well-dialyzed children. AB - BACKGROUND: We have reported catch-up growth with hemodialysis (HD) of approximately 15 hours/week. Without an equilibrated post-treatment blood urea nitrogen, the variable-volume single-pool (VVSP) model will not account for urea rebound, inflating the estimated HD dose (K(d)t/V). A two-pool model (FVDP) predicts rebound, but requires fixed compartment volumes for the equations to be solvable in closed form, also inflating K(d)t/V. METHODS: We developed an approximate perturbation solution (WKB method) to a variable volume, two-pool (VVDP) model. Estimated model parameters were compared with the results of equilibrated kinetic studies using measured clearance K(d) (N = 17). Once the model was validated, we re-analyzed 292 kinetic studies from our earlier cohort, which was considered well-dialyzed on the basis of growth rates (N = 12, mean annual change in height standard deviation score +0.31, mean follow-up of 26 months). RESULTS: For the VVSP, FVDP, and VVDP models, respectively, the mean errors were (1) K(d)t/V, 0.22 +/- 0.07, 0.29 +/- 0.17, 0.06 +/- 0.07 (ANOVA, P < 0.001); (2) urea distribution volume vol/wt (%), -8.2 +/- 4.2, -9.1 +/- 3.0, -2.2 +/- 3.6 (P < 0.001). Sequential studies confirmed reproducibility, with a coefficient of variation < or = 5%. In the earlier cohort, a comparison of the VVSP and VVDP models yielded the following: (1) K(d)t/V, 1.91 +/- 0.35 vs. 1.76 +/- 0.33 (P < 0.001); (2) normalized protein catabolic rate (nPCR, g/kg/day), 1.56 +/- 0.39 vs. 1.52 +/- 0.38 (P < 0.001); and (3) K(d) (whole blood, mL/kg/min), 4.8 +/- 0.9 vs. 4.4 +/- 0.8 (P < 0.001). CONCLUSION: This VVDP model yields reliable estimates of K(d)t/V and other kinetic parameters using standard blood urea nitrogen sampling. Analysis of patients previously characterized as well-dialyzed on the basis of growth rates clarifies the HD dose needed to sustain normal growth. PMID- 11044236 TI - Importance of blood pressure control in hemodialysis patient survival. AB - BACKGROUND: In the general population, hypertension is the leading cause of cardiovascular mortality. In dialysis patients, however, the relationship between blood pressure (BP) and mortality is controversial. We analyzed this relationship in hemodialysis (HD) patients. METHODS: The study population included 405 patients who had survived at least two years on HD. The observation period was initiated at the beginning of the third year. Predialysis BP measurements of all the dialysis treatments performed during the second year of HD was collected as the baseline data. Mean systolic BP (SBP) and mean diastolic BP (DBP) were calculated. Demographic and comorbidity data were collected at the start of the observation period (beginning of third year of HD). Mortality was analyzed at the end of the follow-up (death or December 31, 1998; total mortality), during the first two years of follow-up (years 3 and 4 of HD; early mortality) and after the second year of follow-up (> or = 5 years of HD; late mortality). RESULTS: In the multivariate analysis, SBP and DBP were significantly associated with death. The adjusted total mortalities were U shaped. When early mortality was analyzed, only low BP (DBP <74.5 mm Hg) was significantly associated with mortality. When late mortality was analyzed, only high BP (SBP> 160 mm Hg) was significantly associated with mortality. In the early deaths, a cardiac cause was significantly less frequent, while withdrawal and malignancy were more frequent than in late deaths. CONCLUSIONS: This study confirms that hypertension is a risk factor for mortality in HD patients, and shows the importance of the length of the follow-up time to demonstrate this relationship. The low frequency of a cardiac cause in the early death group suggests that the association between hypotension and mortality in HD patients is not related to cardiovascular causes, and only reflects the association between hypotension and other severe medical conditions. PMID- 11044237 TI - Mixed predilution and postdilution online hemodiafiltration compared with the traditional infusion modes. AB - BACKGROUND: On postdilution hemodiafiltration (post-HDF), convective removal of medium-high molecular weight solutes is, at the highest ultrafiltration rates, limited by high blood viscosity and protein concentration. Prefilter reinfusion (pre-HDF) may overcome this problem, but plasma dilution may affect the overall efficiency of the technique. In this study, an experimental system of online HDF with combined prefilter and postfilter infusion (mixed HDF) was evaluated and compared with the traditional predilution and postdilution modes. METHODS: Removal of urea (U), creatinine (Cr), phosphate (Phos), and beta(2)-microglobulin (beta(2)m), ultrafiltration coefficients of the dialyzer (K(UF)), and rheologic conditions of the blood circuit were evaluated during the three infusion modes (a total of 36 runs lasting 180 min), performed with a polysulfone hemofilter 1.8 m(2), blood flow (Q(b)) 400 mL/min, dialysate flow (Q(d)) 700 mL/min, and infusion rate 120 mL/min (pre-HDF and post-HDF), or 60 + 60 mL/min (mixed HDF). RESULTS: The mean effective U and Cr clearances and urea index of dialysis dose (eKt/V) were significantly higher on post-HDF than on pre-HDF (K(WB) (U) 210 vs. 193 mL/min, K(DQ) (Cr) 152 vs. 142 mL/min, eKt/V 1.41 vs. 1.30), while mixed HDF did not show significant differences versus post-HDF (K(WB) (U) 201 mL/min, K(DQ) (Cr) 149 mL/min). K(DQ) for Phos and beta(2)m were higher on post-HDF in only absolute values. Similar differences were found for instantaneous dialyzer clearances (K(I)) at 60, 120, and 180 minutes of the sessions, with a common trend to decrease with time. K(UF) and the apparent beta(2)m sieving coefficient showed their lowest values toward the end of post-HDF sessions. Increasing filtration fractions (FFs) were associated with increasing transmembrane pressure (TMP) and solute clearances up to FF values of 0.45. These were values achieved in only post-HDF, at which point the curve of the relationship between TMP and FF assumed its steepest exponential trend. CONCLUSIONS: Mixed HDF, by better preserving the characteristics of water and solute transport of the membrane, ensured safer operating conditions than post-HDF, while achieving similar removal of small- and large-size solutes. Optimizing the ratio of prefilter/postfilter infusion and the total infusion according to the relationships found in our study between solute clearances, FF, and TMP, convective flux and transport may avoid excessive hemoconcentration and dangerous pressure gradients. PMID- 11044238 TI - Ischemia-reperfusion injury in renal transplantation is independent of the immunologic background. AB - BACKGROUND: Adhesion molecule expression is important to early transplant failure. However, whether or not adhesion molecule-facilitated inflammation is antigen-dependent is unknown. We tested this hypothesis. METHODS: Rat renal grafts were four-hours cold-preserved in University of Wisconsin (UW) solution, transplanted to syngeneic or allogeneic recipients, and harvested after 2, 6, 12, 24, and 48 hours and after 1 week. The first allogeneic group receive no immunosuppression; two additional groups received either low (1.5 mg/kg) or standard (5 mg/kg) cyclosporine A (CsA). Renal function and morphology were determined; frozen sections were immunostained for P-selectin, L-selectin, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), platelet endothelial cell adhesion molecule-1 (PECAM-1), leukocyte function associated molecule-1 (LFA-1), very late antigen-4 (VLA-4), as well as for neutrophils and monocytes. RESULTS: Selectins increased rapidly at 2 hours and quickly decreased by 12 hours. While P-selectin was expressed on vasculature, L-selectin was found on inflammatory cells. Neutrophil influx and that of LFA-1 positive cells occurred early, peaked between 12 and 24 hours, and paralleled the maximal impairment in renal function. ICAM-1 and PECAM-1 showed similar kinetics and a diffuse distribution. VCAM-1 increased more slowly after 12 hours, peaked at 24 hours, and was localized predominantly on the endothelium of elastic vessels. Between 24 hours and 1 week, all grafts progressively developed dense VLA-4-positive monocytic infiltrates adjacent to vessels expressing VCAM-1. Functional, morphological, and immunohistochemical parameters did not differ between isografts and allografts at one week. However, by day 10, allografts showed severe vascular and cellular rejection, while injury in isografts resolved. Immunosuppression with CsA did not reverse the inflammation induced by ischemia-reperfusion injury. CONCLUSIONS: The early inflammation after ischemia reperfusion injury is largely independent of the immunologic background. We suggest that initial injury prevention should receive the highest priority. PMID- 11044239 TI - Factors associated with the prevalence of arteriovenous fistulas in hemodialysis patients in the HEMO study. Hemodialysis (HEMO) Study Group. AB - BACKGROUND: Arteriovenous (AV) fistulas are the vascular access of choice for hemodialysis patients, but only about 20% of hemodialysis patients in the United States dialyze with fistulas. There is little information known about the factors associated with this low prevalence of fistulas. METHODS: Multiple logistic regression analysis was used to evaluate the independent contribution of factors associated with AV fistula use among patients enrolled in the HEMO Study. The analysis was conducted in 1824 patients with fistulas or grafts at 45 dialysis units (15 clinical centers). RESULTS: Thirty-four percent of the patients had fistulas. The prevalence of fistulas varied markedly from 4 to 77% among the individual dialysis units (P < 0.001). Multiple regression analysis revealed five demographic and clinical factors that were each independently associated with a lower likelihood of having a fistula, even after adjustment for dialysis unit. Specifically, the prevalence of fistulas was lower in females than males [adjusted odds ratio (AOR) 0.37, 95% CI, 0.28 to 0.48], lower in patients with peripheral vascular disease than in those without (AOR 0.55, 95% CI, 0.38 to 0.79), lower in blacks than in non-blacks (AOR 0.64, 95% CI, 0.46 to 0.89), lower in obese patients (AOR per 5 kg/m(2) body mass index, 0.76, 95% CI, 0.65 to 0.87), and lower in older patients (AOR per 10 years, 0.85, 95% CI, 0.78 to 0.94). The differences in the prevalence of fistulas among the dialysis units remained statistically significant (P < 0.001) after adjustment for these demographic and clinical factors. Finally, there were substantial variations in the prevalence of fistulas even among dialysis units in a single metropolitan area. CONCLUSIONS: Future efforts to increase the prevalence of fistulas in hemodialysis patients should be directed at both hemodialysis units and patient subpopulations with a low fistula prevalence. PMID- 11044240 TI - Glutathione S-transferase polymorphisms and skin cancer after renal transplantation. AB - BACKGROUND: Susceptibility to skin cancer after transplantation is multifactorial, and risk factors include skin type, sun exposure, and level of immunosuppression. A major mechanism of carcinogenesis is ultraviolet radiation induced free radical damage, and genetically determined ability to metabolize free radicals may also predispose to skin cancer. The glutathione S-transferase enzymes play a major role in limiting the toxic effects of reactive oxygen species, and this study was designed to determine whether polymorphisms in these enzymes are associated with skin cancers in renal transplant recipients. METHODS: Two hundred twenty-two long-term survivors of renal transplantation were examined for polymorphisms in the GSTM1, GSTT1, and GSTP1 genes, using a unified polymerase chain reaction with sequence specific primers (PCR-SSP) genotyping method. RESULTS: The GSTP1*C allele was associated with the development of squamous cell carcinomas (SCCs; P = 0.01). No associations of the GSTM1 null genotype or the GSTT1 null genotype were identified, and the development of basal cell carcinomas was not associated with any GST polymorphism studied. CONCLUSIONS: These results indicate that genetic variation in enzymes involved in free radical metabolism in the skin are associated with the development of skin cancer. While all renal transplant recipients should be advised to protect themselves from the sun, the identification of transplant patients with a genetic predisposition to skin tumors may permit the targeting of preventative and early intervention strategies to high-risk individuals. PMID- 11044241 TI - Spurious estimations of sodium removal during CAPD when [Na](+) is measured by Na electrode methodology. AB - BACKGROUND: The aim of this study was to investigate the effect of pH and glucose concentration on sodium removal and the dialysate and plasma sodium ratio (D/PNa) as measured by means of a flame photometer (NaF) or direct ion-selective electrode (NaE) in continuous ambulatory peritoneal dialysis (CAPD). METHODS: In vitro, glucose concentration, pH, NaF, and NaE were measured in fresh peritoneal dialysis solutions (PDSs) before and after the addition of glucose or KOH. In vivo, 66 four-hour peritoneal equilibration tests were performed in 35 patients on CAPD using a low pH PDS with a glucose concentration of 3.86%. RESULTS: In vitro, NaF and NaE were significantly influenced by the glucose concentration and pH of the PDS. In vivo, in fresh PDS, there was a significant difference between the NaF and NaE results; the respective median values were 132.1 (interquartile range 129.3 to 137.5) versus 138.0 (134.4 to 141.5) mmol/L (P < 0.0001). The D/PNa ratio calculated by NaE was significantly lower than that calculated by NaF (0.88 +/- 0.03 vs. 0.91 +/- 0.04 and 0. 90 +/- 0.03 vs. 0.94 +/- 0.04 at 60 and 240 min, respectively, P < 0.0001), whereas there was no significant difference between the NaE and NaF values after correction for plasma water and a Donnan factor of 0.96 (0.88 +/- 0.03 vs. 0.88 +/- 0.04 and 0.90 +/- 0.03 vs. 0.91 +/- 0.04, P < 0.3473). Sodium removal was significantly lower when calculated as NaE than when calculated as NaF (43.9 +/- 32.7 vs. 61.0 +/- 32.2 mmol, P < 0.0001). CONCLUSIONS: The fresh PDS sodium concentration can be corrected using a glucose concentration-related factor. The D/PNa ratio calculated as NaE or NaF is not different after correction for plasma water and a Donnan factor of 0.96. Sodium removal must be measured by means of NaF rather than NaE. This could have an important clinical impact. PMID- 11044242 TI - Risk factors for hip fracture among patients with end-stage renal disease. AB - BACKGROUND: Although bone disease is well described among end-stage renal disease (ESRD) patients, little attention has been paid to the occurrence of fracture. We sought to identify factors that are associated with hip fracture among ESRD patients. METHODS: Data from patients who participated in the United States Renal Data System Dialysis Morbidity and Mortality Study Wave 1 were used for this study. Hip fractures occurring among these patients between 1993 and 1996 were identified from Medicare claims data available from the United States Renal Data System. Cox proportional hazards models were used to estimate the risk of hip fracture associated with demographic and medical variables. RESULTS: Of the 4952 patients included in this analysis, 103 sustained a hip fracture. In the multivariate analysis, age (per increasing decade, RR = 1.40, 95% CI 1.20, 1.64), female gender (RR = 2.26, 95% CI 1.48, 3.44), race (blacks compared with whites, RR = 0.58, 95% CI 0.37, 0.91), body mass index (per 1 unit increase, RR 0.89, 95% CI 0.86, 0.93), and the presence of peripheral vascular disease (RR 1.94, 95% CI 1.29, 2.92) were independently associated with hip fracture. Serum intact parathyroid hormone (iPTH), aluminum, diabetes, and bicarbonate levels did not appreciably influence the risk of hip fracture. CONCLUSIONS: Demographic and other characteristics that predict risk of hip fracture in the population at large also do so in ESRD patients. However, we could identify no characteristics of ESRD or its treatment that were independently related to hip fracture incidence. PMID- 11044243 TI - Acute rejection-associated tubular basement membrane defects and chronic allograft nephropathy. AB - BACKGROUND: Acute rejection is a major risk factor for chronic allograft nephropathy, although the link(s) between these events is not understood. The hypothesis of this study is that alterations in tubular basement membranes (TBMs) that occur during acute rejection may be irreversible and thereby play a role in the development of chronic allograft nephropathy. METHODS: Fourteen renal transplant patients were selected, each having had two or more biopsies performed (42 total). All biopsies were scored for acute and chronic rejection using Banff 1997 criteria. The initial biopsy showed only acute interstitial rejection (type I rejection). No biopsies contained significant chronic arterial lesions of chronic vascular rejection. The entire cortex was examined on Jones methenamine silver-stained sections at x400 for interruption in TBM staining. The number of tubules with TBM abnormalities was counted, and the renal cortical area was measured by image analysis. Periodic acid-Schiff/immunoperoxidase stain was performed on 12 acute rejection biopsies stained for laminin, cytokeratin 7, CD3, CD20, and CD68. Controls consisted of 11 biopsies (8 negative for rejection and 3 acute tubular necrosis). RESULTS: Numerous TBM alterations in silver staining were identified as being associated with acute rejection and tubulitis, consisting of abrupt TBM discontinuities and/or extreme attenuation with segmental or complete absence of TBM. A loss of TBM matrix proteins was confirmed by absent laminin staining in areas of acute rejection and tubulitis. There was herniation of tubular cells into the interstitium through TBM defects confirmed by cytokeratin staining. The TBM defects were spatially associated with inflammatory cells, particularly macrophages. When the biopsies were divided into two groups, <10 and> 10 TBM breaks/mm2, there were statistically significant morphologic and clinical correlations. The number of TBM disruptions correlated with the serum creatinine at the time of biopsy, a combined Banff t + i score, the difference in tubular atrophy between the initial and most recent biopsy and the difference between the nadir creatinine and most recent creatinine. CONCLUSION: Damage to TBM develops in acute rejection as a consequence of interstitial inflammation and tubulitis. These lytic events correlate with the later development of clinical and morphologic evidence of chronic injury in the absence of arterial injury of chronic rejection. We suggest that chronic allograft nephropathy may have an inflammatory interstitial origin. PMID- 11044244 TI - CD28 knockout mice as a useful clue to examine the pathogenesis of chronic graft versus-host reaction. AB - BACKGROUND: Injection of BALB/c or DBA/2 spleen cells into F1 C57BL/6 (B6) hybrids induces a graft-versus-host reaction (GVHR) of a chronic stimulatory type that results in clinical and pathologic manifestations that resemble the human systemic lupus erythematosus (SLE). The aim of the present study was to examine the role of a major T-cell costimulatory signal receptor, CD28, in the production of autoantibody and the development of an immune complex glomerulonephritis, which are common in SLE pathology. METHODS: For this purpose, CD28-deficient (CD28KO) mice were used for the source of donor lymphocytes. Chronic GVHR was induced by an injection of BALB/c or BALB. CD28KO donor cells into normal BCF1 mice. Serum titers of anti-dsDNA antibodies were assessed by enzyme-linked immunosorbent assay (ELISA) and major histocompatibility complex (MHC) class II antigen expression on B cells were tested by flow cytometry. In addition, depositions of immunoglobulin (Ig) were examined by direct immunofluorescence staining on frozen kidney sections. RESULTS: When (BALB/c x B6)F1 mice were injected with parental BALB/c lymphocytes, serum anti-dsDNA titer was significantly increased in association with nonspecific B-cell activation and IgG deposition in the glomerular basement membrane. In sharp contrast, none of these signs were observed in F1 mice, which were injected with CD28KO spleen cells. CONCLUSION: The CD28-mediated T-cell costimulatory pathway plays a pivotal role in the development of polyclonal B-cell activation, autoantibody production, and an immune complex glomerulonephritis. We propose that CD28KO mice are useful clues in examining the pathogenesis of experimental lupus nephritis. PMID- 11044245 TI - Initial clinical results with the LifeSite Hemodialysis Access System. AB - BACKGROUND: The LifeSite Hemodialysis Access System is a subcutaneous valve with an internal pinch clamp that is actuated with a standard 14-gauge dialysis needle, connected to a single lumen cannula placed in the central venous circulation for hemodialysis (HD). METHODS: The LifeSite System (2 valves) was implanted in 23 patients with immediate dual-needle HD use. The cannulas were placed in either the jugular or the subclavian veins and were connected to the subcutaneous valves located in the upper chest area. RESULTS: The mean duration of device survival for the LifeSite System was 6.8 +/- 0.97 months. During this period, the device achieved prescription HD blood flow rates averaging 384.7 +/- 78.5 mL/min with a venous pressure of 223.2 +/- 60.3 mm Hg. After 125 patient months, device removal because of infection was at a rate of 2.5 per 1000 days, and there were no devices removed because of poor flow. The average Kt/V for these patients was 1.51. CONCLUSION: This preliminary clinical study has validated the applicability of the LifeSite Hemodialysis Access System as an access for HD. It is easily implanted and easily used, provides safe and effective dialysis, and is well accepted by patients. It should be especially useful as a bridge device to allow for maturation of a native fistula and will provide an alternative for long-term use in patients in whom a peripheral dialysis access is not feasible. PMID- 11044246 TI - Mass spectrometric monitoring of albumin in uremia. AB - BACKGROUND: Advanced glycation end products (AGEs) are a novel class of uremic toxins. In plasma, they are present in proteins and also in low molecular mass peptides. AGE-modified peptides are thought to bind and modify plasma proteins. Monitoring of the consequent increase in molecular mass of serum albumin may be used in surveillance of the clinical management of uremia. METHODS: We investigated molecular mass changes of human serum albumin (HSA) glycated by glucose and methylglyoxal in vitro and of subjects with moderate renal impairment, end-stage renal disease (ESRD), ESRD on hemodialysis, and normal healthy controls by matrix-assisted laser desorption ionization mass spectrometry. RESULTS: Fatty acid-free HSA had a molecular mass of 66,446 +/- 114 D. Mean (+/-SD) molecular mass increases were HSA minimally glycated by glucose 399 +/- 88 D (N = 5, P < 0.001), HSA highly glycated by glucose 6780 +/- 122 D (N = 5, P < 0.001), HSA minimally glycated by methylglyoxal 73 +/- 121 D (N = 5, P > 0.05), and HSA without fatty acid removal 535 +/- 90 D (N = 5, P < 0.001). For HSA of human subjects, mean (+/- SD) molecular mass increases were normal healthy controls 243 +/- 97 D (N = 5), moderate renal impairment 350 +/- 83 D (P > 0.05 with respect to controls, N = 5), ESRD 498 +/- 128 (P < 0.02 with respect to controls, N = 3), and ESRD on hemodialysis 438 +/- 85 D (P < 0.02 with respect to controls, N = 5). The mean molecular mass of albumin of all groups was increased significantly with respect to that of fatty acid free albumin (P < 0.001). CONCLUSIONS: Only ESRD was associated with a significant increase in the molecular mass of HSA in vivo. Since this mass increase was very low and much lower than reported for AGE-modified peptides, it may reflect AGE formation on HSA by alpha-oxoaldehydes that accumulate in uremia, rather than modification of albumin by AGE-modified peptides. The molecular mass of HSA in vivo was indicative of a minimal and not a high extent of glycation. PMID- 11044247 TI - Renal cell loss through cell suicide. PMID- 11044248 TI - BMP receptors in kidney. PMID- 11044249 TI - Endothelin antagonism and contrast nephropathy. PMID- 11044250 TI - Reply from the authors PMID- 11044251 TI - Lupus nephritis and the anti-phospholipid antibody syndrome in pregnancy. PMID- 11044252 TI - Role of Sertoli cells in injury-associated testicular germ cell apoptosis. AB - This review examines experimental models of Sertoli cell injury resulting in germ cell apoptosis. Since germ cells exist in an environment created by Sertoli cells, paracrine signaling between these intimately associated cells must regulate the process of germ cell death. Germ cell apoptosis may be signaled by a decrease in Sertoli cell pro-survival factors, an increase in Sertoli cell pro apoptotic factors, or both. The different models of Sertoli cell injury indicate that spermatogenesis is susceptible to disruption, and that targeting critical Sertoli cell functions can lead to rapid and massive germ cell death. PMID- 11044253 TI - Sugar transport regulation: comparative characterization of the effect of NADH CoQ reductase deficiency in two cell culture systems. AB - In this report, we have characterized the upregulation of glucose transport in two different respiration-deficient fibroblast cell cultures. We have demonstrated that glucose transport increases in respiration-deficient cells as measured by 2 deoxy D-glucose transport and is readily observed in both the WG750 human and G14 Chinese hamster fibroblast respiration-deficient cell lines when compared with the MCH55 normal human and V79 parental Chinese hamster cell lines, respectively. Using subcellular fractionation techniques, the GLUT 1 glucose transporter was found located predominantly in the plasma membrane-enriched fraction of the human and hamster cell lines. In human cells, the expression of the GLUT 1 glucose transporter was elevated three-fold in the plasma membrane enriched fraction of the WG750 respiration-deficient mutant cells. In the Chinese hamster cell lines, the respiration-deficient G14 cells exhibited no such GLUT 1 glucose transporter elevation in the plasma membrane-enriched fraction, yet expressed a >2-fold increase in glucose transport. Furthermore, the G14 cells had a similar content of GLUT 1 glucose transporter in the plasma membrane fraction when compared with the V79 parental cell line. Using Western blot analysis, the GLUT 1 glucose transporter in G14 cells exhibited a different mobility on a polyacrylamide gel when compared with the mobility of the GLUT 1 glucose transporter of the V79 cell line. This differential mobility of the glucose transporters in the hamster cells appeared to be related to glycosylation differences of the glucose transporters. Although normal human and hamster cell lines exhibited significant increases in insulin-stimulated sugar transport (P < 0.05), the two respective respiration-deficient cell lines exhibited no significant increases in insulin-stimulated sugar transport (P > 0.05). Additionally, the expression of the GLUT 1 mRNA in the human WG750 mutant cells was elevated when compared with GLUT 1 mRNA in normal cells. Insulin exposure significantly increased GLUT 1 mRNA in human cells (P < 0.05). No differences in the GLUT 1 mRNA were observed between both hamster cell lines. Thus, both respiration-deficient cell lines are insulin resistant (i.e., regarding their insulin-stimulated sugar transport). The respiration-deficient mutation results in an increased sugar transport in the human and hamster cells; however, the human cells adapt to the mutation by increasing their levels of GLUT 1 mRNA and eventually membrane-located glucose transporters. On the other hand, the hamster cells adapt by apparently modifying their glucose transporters' intrinsic activity via glycosylation. We feel that these cell systems can be effective models to study the multiple factors involved in sugar transport regulation in vertebrate cells. PMID- 11044254 TI - Age-related changes in rat hepatic acetyl-coenzyme A carboxylase. AB - Acetyl-CoA carboxylase (ACC) catalyzes the rate-limiting step in the synthesis of long-chain fatty acids. Since aging influences adiposity, we studied the activity of ACC and its mRNA content in livers of 4-, 12-, and 24-month-old male Fischer 344 rats. The mean (+/- SEM) activity of ACC (mU/mg protein) in liver homogenates from 4-month-old rats was 1.01 +/- 0.14. There was an 80% increase in activity (1.83 +/- 0.27) in 12-month-old rats (P < 0.01). However, there was significantly less activity (0.46 +/- 0.06) in livers of 24-month-old rats (P < 0.001). The total activity of ACC (per g liver) followed the same trend. The enzyme from all age groups was purified by avidin-affinity chromatography. The purified preparation migrated as a major protein band (M(r) 262,000) on sodium dodecyl sulfate (SDS)-polyacrylamide gels. The specific activity of the purified preparation was 1.5, 1.8, and 1.8 U/mg for 4-, 12-, and 24-month-old rats, respectively. The alkali-labile phosphate content was 5.66 +/- 0.17, 5.64 +/- 0.21, and 6.21 +/- 0.35 mols P(i)/mole subunit for 4-, 12-, and 24-month-old rats, respectively. These age-related differences were not significant. The hepatic ACC mRNA measured by ribonuclease protection assay when corrected for G3PDH mRNA was significantly reduced in 24-month-old rats (0.24 +/- 0.03) compared with 12-month-old (0.58 +/- 0.04) or 4-month-old rats (0.43 +/- 0.007) P < 0.01. In summary: (i) Aging in rats is associated with significant changes in ACC activity; (ii) the purified ACC preparations from the three age groups had similar specific activity and similar phosphate content; and (iii) the changes in ACC mRNA content of the liver paralleled the changes in total enzyme activity when 12-month-old rats were compared with 24-month-old rats whereas the increase in ACC activity in 12-month-old rats compared with 4-month-old rats could not be ascribed to changes in hepatic mRNA levels. These results indicate that the age related changes in hepatic ACC occur at a post-translational level during early years of aging and at a pretranslational level at late states of senescence. These changes may contribute to the age-related alterations in body adiposity. PMID- 11044255 TI - Induction of hepatic insulin-like growth factor binding protein-1 (IGFBP-1) in rats by dietary n-6 polyunsaturated fatty acids. AB - The insulin-like growth factors (IGFs) are mitogenic polypeptides that have been linked to a variety of normal physiological processes as well as neoplasia. Overexpression of several components of the IGF system is associated with hepatocarcinogenesis in humans and rodents. In rat liver, diets rich in n-6 polyunsaturated fatty acid (PUFA) enhance the development of preneoplastic lesions and tumors. Therefore, the objective of this study was to determine the effect of these dietary fatty acids on the hepatic expression of the various components of the IGF system. The mRNA levels of IGF-1 and the type 1 receptor were not different in livers of rats fed a diet containing 20% corn oil (CO) compared with those fed 5% CO. Analysis of the IGF binding proteins revealed that insulin-like growth factor binding protein-1 (IGFBP-1) levels were altered by the amount and type of dietary fat. A 2.5-fold induction of IGFBP-1 mRNA occurred within 1 week after the animals were fed the 20% corn oil diet compared with those fed 5% CO and was further enhanced to over 6-fold after 1 month. Furthermore, IGFBP-1 protein was only detectable in the livers of animals fed the 20% CO diet. Induction of IGFBP-1 mRNA (4.5-fold) also occurred in rats fed a high-fat diet containing safflower (rich in n-6 PUFAs) compared with those fed a high-fat diet containing menhaden oil (rich in n-3 PUFAs). The induction of IGFBP 1 mRNA was independent of serum insulin levels and the development of insulin resistance. Since IGFBP-1 mRNA is upregulated in regenerating liver, we reasoned that the induction of IGFBP-1 mRNA may be associated with an increase in cell proliferation; however, no difference was observed in the hepatic labeling index of rats fed the 20% CO compared with the 5% CO diet. In summary, these studies show a striking induction by dietary n-6 PUFAs of hepatic IGFBP-1, a protein that has been implicated in liver cancer development. PMID- 11044256 TI - Anti-inflammatory effects of ergotamine in steers. AB - The objective of this experiment was to investigate whether the ergot alkaloid, ergotamine (ET), an alkaloid used to model fescue toxicosis in cattle, modifies the response of cattle to endotoxin (LPS) challenge. Steers (n = 16) were divided into the following treatment groups: control (C), ergotamine (ET), endotoxin (LPS), and ET + LPS. ET and ET + LPS groups received a single bolus intravenous injection of ET (40 microg. kg. body wt(-1)), whereas C and LPS steers received a single bolus injection of sterile vehicle. Thirty minutes after ET/vehicle administration, a single bolus intravenous injection of LPS (0.2 microg. kg. body wt(-1)) was given. Blood was collected at various time points for 48 hr post. Endotoxin increased rectal temperature (RT) and the circulating levels of tumor necrosis factor-alpha (TNF-alpha), cortisol, haptoglobin (Hp), thromboxane B(2) (TXB(2)). The circulating Hp, TNF-alpha, and TXB(2) increases were blunted by pretreatment with ET compared with ET + LPS. Ergotamine by itself increased circulating cortisol and RT, whereas it decreased serum prolactin (PRL). Therefore, whereas administration of LPS at 0.2 microg/kg to steers resulted in an expected response, the combination of ET + LPS attenuated major effects of LPS alone. Thus, acute administration of ET appeared to be anti-inflammatory as it decreased the inflammatory response to LPS, an effect likely driven at least in part by the ET-caused cortisol increase. PMID- 11044257 TI - Cell death induction by CTL: perforin/granzyme B system dominantly acts for cell death induction in human hepatocellular carcinoma cells. AB - Cell death induction by cytotoxic T lymphocytes (CTLs) is an important thesis for the understanding of tumor immunotherapy. In the current study we investigated the molecular machinery of CTL-induced cell death in human hepatocellular carcinoma cell lines (HCC lines). CTLs prepared from human peripheral blood induced cell death in all tested HCC lines. As the CTL-induced death system, the effectiveness of Fas ligand/Fas and/or Perforin/Granzyme B systems has been suggested, whereas cell death induction by CTLs was shown independently on Fas expression in the current study. Using various tetrapeptide inhibitors for caspase and its associated factor, we additionally demonstrated that inhibitors for caspase 3 (Ac-DEVD-CHO) and caspase 8/granzyme B (Ac-IETD-CHO) suppressed CTL induced cell death, but an inhibitor for Fas-activated serine proteinase, which acts for the caspase 3 activator, did not, suggesting that CTL-induced cell death was initiated by the Perforin/Granzyme B system, rather than the Fas ligand/Fas system. On the basis of our current results, we report here that the Perforin/Granzyme B system acts dominantly for the cell death induction of HCC lines. PMID- 11044259 TI - 3rd international symposium on Cell/Tissue injury and Cytoprotection/Organoprotection PMID- 11044258 TI - Role of nitric oxide and superoxide in acute cardiac allograft rejection in rats. AB - The role of NO and superoxide (O(2)(-)) in tissue injury during cardiac allograft rejection was investigated by using a rat ex vivo organ perfusion system. Excessive NO production and inducible NO synthase (iNOS) expression were observed in cardiac allografts at 5 days after cardiac transplantation, but not in cardiac isografts, as identified by electron spin resonance spectroscopy and Northern blotting. Cardiac isografts or allografts obtained on Day 5 after transplantation were perfused with Krebs bicarbonate buffer with or without various antidotes for NO or O(2)-, including N(omega)-monomethyl-L-arginine (L-NMMA; 1 mM), 2-phenyl 4,4,5, 5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO; 100 microM), 4-amino-6 hydroxypyrazolo[3,4-d]pyrimidine (AHPP; a xanthine oxidase inhibitor; 100 microM), and superoxide dismutase (SOD; 100 units/ml). Treatment of the cardiac allografts with PTIO showed most remarkable improvement of the cardiac injury as revealed by significant reduction in aspartate transaminase, lactate dehydrogenase, and creatine phosphokinase concentrations in the perfusate. Similar but less potent protective effect on the allograft injury was observed by treatment with L-NMMA, AHPP, and SOD. Immunohistochemical analyses for iNOS and nitrotyrosine indicated that iNOS is mainly expressed by macrophages infiltrating the allograft tissues, and nitrotyrosine formation was demonstrated not only in macrophages but also in cardiac myocytes of the allografts, providing indirect evidence for the generation of peroxynitrite during allograft rejection. Our results suggest that tissue injury in rat cardiac allografts during acute rejection is mediated by both NO and O(2)(-), possibly through peroxynitrite formation. PMID- 11044260 TI - Improving the performance of health systems: the World Health Report as go between for scientific evidence and ideological discourse. PMID- 11044261 TI - Sociodemographic characteristics, care, feeding practices, and growth of cohorts of children born to HIV-1 seropositive and seronegative mothers in Nairobi, Kenya. AB - OBJECTIVES To compare sociodemographic profiles, child care, child feeding practices and growth indices of children born to HIV-1 seropositive and seronegative mothers. METHODS: A cohort study of 234 children (seropositive and seronegative) born to HIV-1 seropositive mothers and 139 children born to seronegative mothers in Pumwani Maternity Hospital which serves a low-income population in Nairobi, Kenya from December 1991 and January 1994. RESULTS: With few exceptions, at the time of their birth children in all three cohorts had parents with similar characteristics, lived in similar housing in similar geographical areas, had their mothers as their primary care givers, had similar feeding practices and similar growth status and patterns. However, the HIV-1 seropositive mothers were slightly younger (23.8 years vs. 25.0 years, P < 0.01), if married they were less likely to be their husband's first wife (79% vs. 91%, P = 0.02) and more likely to have a one-room house (75% vs. 63%, P = 0.04). All three cohorts had mean Z-scores in length-for-age and in weight-for-height within the normal range (>/= 2.0 Z-scores) from birth to 21 months with the exception of the length-for-age of the seropositive children at the 18-month visit. In all cohorts length-for-age became more compromised than weight-for-length, dropping to about -1.45 Z-score by 21 months; in contrast, weight-for-length dropped to about -0.5 Z-score by this age. The only statistically significant differences in growth indices among the three cohorts were between the two cohorts of seronegative children: those with seronegative mothers were less compromised in length-for-age at 1.5 months (mean Z-score = -0.19 vs. -0.48, P < 0.05) and more compromised in weight-for-length at 6 months (mean Z-score = 0.10 vs. 0.45, P < 0.05) and at 18 months (mean Z-score = -0.73 vs. -0.16, P < 0.05). 27-34% were exclusively breastfed at 1.5 months; 52-61% consumed solid foods in addition to breast milk by 2.5 months. CONCLUSIONS: Low-income HIV-1 seropositive- and seronegative-born children were from families with similar characteristics and similar housing environments. Similar growth patterns in the cohorts suggest that the challenging environment and the choice of weaning foods had an impact on all three cohorts. The aggressive care given the children with HIV-1 seropositive mothers and their children may have reduced the progression and impact of HIV-1 disease on the growth of the seropositive children. Further research is needed to corroborate our findings to be certain that our results are not affected by loss to follow-up bias: we lost the same proportion in all three cohorts but cannot verify that the children we lost had the same growth patterns as those who remained in the study. PMID- 11044262 TI - AIDS caused by HIV1 and HIV2 infection: are there clinical differences? Results of AIDS surveillance 1986-97 at Fann Hospital in Dakar, Senegal. AB - OBJECTIVE: To compare the clinical manifestations observed in AIDS patients infected with HIV2 and HIV1 infection. METHODS: The medical records of AIDS patients hospitalized between January 1986 and July 1997 at the Department of Infectious Diseases of Fann Hospital, Dakar, were reviewed. RESULTS: 599 hospitalizations (76%) were HIV1 seropositive patients, 137 (17%) were HIV2 seropositive patients and 54 (7%) were patients serologically dually reactive to HIV1 and HIV2. There was no significant difference in medium CD4 lymphocyte count between patients with HIV1 and HIV2 infection. Chronic diarrhoea and diarrhoea caused by bacterial infections were more frequently observed in HIV2-infected individuals. Oral candidiasis and chronic fever were more often noted in patients with HIV1 infection. Bacterial and cryptococcal meningitis was only observed among patients with HIV1 infection. CONCLUSIONS: Certain clinical differences were observed comparing AIDS patients with HIV1 and those with HIV2 infection. As there is no clear physiopathological explanation for these differences, additional studies with larger numbers of AIDS patients are needed to determine whether these differences are real. PMID- 11044263 TI - Chloroquine- and sulfadoxine-pyrimethamine-resistant Falciparum malaria in vivo - a pilot study in rural Zambia. AB - BACKGROUND: Chloroquine (CQ) and Sulfadoxine-Pyrimethamine (SP) are the predominantly used antimalarials in Zambia and other parts of East Africa, but increasing resistance of P. falciparum is a major concern. METHODS: Seventy consecutive patients with uncomplicated falciparum malaria were enrolled. In 43 patients, no prior CQ use could be demonstrated by history and urianalysis (qualitative test, Dill & Glazko) and these patients were given CQ; the other 27 had taken CQ before and received SP. RESULTS: Combined R-II and R-III CQ resistance was 58% (60% in under-fives), which is the range previously reported from Zambia. By contrast, SP-resistance (R-II and R-III) was much higher (26%) than previously reported (3% - 17%). The history of prior CQ intake correlated well with the results of the Dill-Glazko test; there was no evidence for prior SP intake to explain these results. CONCLUSION: If our findings of SP resistance are confirmed, other drugs such as quinine, atovaquone/proguanil and artemisinin are required to treat malaria in Zambia. PMID- 11044264 TI - Micronutrient and iron supplementation and effective antimalarial treatment synergistically improve childhood anaemia. AB - The control of childhood anaemia in malaria holoendemic areas is a major public health challenge for which an optimal strategy remains to be determined. Malaria prevention may compromise the development of partial immunity. Regular micronutrient supplementation has been suggested as an alternative but its effectiveness remains unsettled. We therefore conducted a randomised placebo controlled intervention trial with 207 Tanzanian children aged 5 months to 3 years on the efficacy of supervised supplementation of low-dose micronutrients including iron (Poly Vi-Sol with iron) three times per week, with an average attendance of >/= 90%. The mean haemoglobin (Hb) level increased by 8 g/l more in children on supplement (95% CI 3-12) during the 5-month study. All age groups benefited from the intervention including severely anaemic subjects. The mean erythrocyte cell volume (MCV) increased but Hb in children >/= 24 months improved independently of MCV and no relation was found with hookworm infection. The data therefore suggest that micronutrients other than iron also contributed to Hb improvement. In the supplement group of children who had received sulfadoxine pyrimethamine (SP) treatment, the mean Hb level increased synergistically by 22 g/l (95% CI 13-30) compared to 7 g/l (95% CI 3-10) in those without such treatment. Supplementation did not affect malaria incidence. In conclusion, micronutrient supplementation improves childhood anaemia in malaria holoendemic areas and this effect is synergistically enhanced by temporary clearance of parasitaemia. PMID- 11044265 TI - 'Killer' canines: the morbidity and mortality of ebino in northern Uganda. AB - In northern Uganda, unerupted primary canine teeth are commonly extracted because they are believed to cause diarrhoea, vomiting, and fever. This practice, known as ebino, is performed under very crude conditions often using unclean tools. To evaluate the morbidity and mortality of complications related to ebino, we retrospectively analysed discharge records from the paediatric ward of Lacor Hospital, Gulu. In the period 1992-98, ebino-related complications, mainly sepsis and anaemia, were among the leading causes of admission (n = 740) and hospital death (n = 156, case fatality rate = 21.1%, proportional mortality rate = 3.3%). Discouraging the adoption of deeply rooted traditional practices that are potentially hazardous to health should be a public health priority in northern Uganda. This could be done by educating not only the general public, but also traditional healers and community and religious leaders, who could convey the knowledge to their people. PMID- 11044266 TI - Antibiotic medication and bacterial resistance to antibiotics: a survey of children in a Vietnamese community. AB - OBJECTIVE: To investigate antibiotic use and antibiotic susceptibility of respiratory tract pathogens in children aged 1-5 years in Bavi, Vietnam. METHOD: Nasopharynx and throat specimens were collected from 200 children from randomly selected households in a demographically defined population. Respiratory isolates were tested for antibiotic susceptibility according to the standard disk diffusion method. A questionnaire survey of carers elicited information on type of antibiotic used, duration of treatment, where the antibiotics had been purchased, type of treatment information retained by carers and episodes of illness preceding the study. RESULTS: 82% of the children had at least one symptom of acute respiratory tract infection (ARI) in the 4 weeks prior to the study, and of these 91% were treated with antibiotics. The most commonly used antibiotics were ampicillin (74%), penicillin (12%), amoxicillin (11%), erythromycin (5%), tetracycline (4%) and streptomycin (2%). Ampicillin was used for 3.3 days on average (SD:1.8) and penicillin for 2.6 days (SD:0.7). When deciding which antibiotic to use, 67% of the carers consulted the pharmacy seller, 11% decided themselves and 22% followed the doctor's prescription. The carrier rate of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis was 50%, 39% and 17%, respectively. Isolates from 145 children were susceptibility tested, and 74% were found to carry resistant pathogens. Of the tested isolates, 90% of S. pneumoniae, 68% of H. influenzae and 74% of M. catarrhalis were resistant to at least one antibiotic. The mean number of antibiotics (susceptible strains excluded) to which resistance was found was 2.0 (SD:1.2), 2.5 (SD:1.8) and 2.1 (SD:0.9), respectively. S. pneumoniae and H. influenzae showed high resistance to tetracycline (88% and 32%, respectively), trimethoprim/sulphonamide (32% and 44%), and chloramphenicol (25% and 24%). 23% of S. pneumoniae were erythromycin-resistant and 18% of H. influenzae isolates were resistant to ampicillin. There was a significant difference in ampicillin and penicillin resistance between the group of children previously treated with beta lactam antibiotics and the group of children who did not receive antibiotics. CONCLUSION: As reported by the carers, children in Bavi are treated with antibiotics frequently. Most antibiotics were obtained without consulting a doctor. High levels of antibiotic resistance and high prevalence of multidrug resistant strains were found among respiratory pathogens. The existence of a large reservoir of resistance genes among children in low-income countries represents a threat to the success of antibiotic therapy throughout the world. Multi-faceted programmes to improve rational use of antibiotics in Vietnam are urgently needed. PMID- 11044268 TI - High seroprevalence of antibodies to varicella zoster virus in adult women in a tropical climate. AB - OBJECTIVES: To determine whether the seroprevalence of antibodies to varicella zoster virus (VZV) in adults is similar to that reported in tropical populations elsewhere. METHODS: We measured the seroprevalence of VZV IgG antibodies, using an enzyme immunoassay (EIA) in women attending an antenatal clinic in an urban centre in tropical Australia. RESULTS: The overall seroprevalence of VZV antibodies in 298 women was 92% (95% CI 88-95), with no difference between women who spent their childhoods in the tropics and colleagues. None of the overseas born women was seronegative. CONCLUSION: The seroprevalence of VZV antibodies in this tropical population in Australia is as high as that reported from temperate regions, suggesting that social and cultural factors and population mobility are more important determinants of age distribution of VZV immunity than tropical climate. PMID- 11044267 TI - Cancer incidence in French Polynesia 1985-95. AB - OBJECTIVE: To describe the variations in cancer incidence in the population born in French Polynesia (FP) according to the archipelago of birth and to compare this incidence with that of Hawaiians and Maoris. METHODS: Study of data from the Cancer Registry of FP, evacuation files, insurance records, hospital and pathology laboratory files. RESULTS: The overall world standardized cancer incidence in FP during the 1985-95 period in the populations born and living in FP was 246 per 105 person-years (PY) among women and 244 per 105 PY among men. The overall cancer incidence was similar to that in Hawaiians, but 25% lower than in Maoris. Digestive tract cancer incidence was a third that of these two reference populations, whereas that of pharynx, larynx and thyroid cancers was approximately twice as high. The overall cancer incidence rate increased between the period 1985-89 and the period 1990-95 in women, but was stable in men. Colorectal cancer incidence was highest in inhabitants born on the Windward Islands. Women born on the Austral Islands had a higher thyroid and liver cancer incidence and a lower breast cancer incidence. CONCLUSIONS: Further studies are needed to elucidate the variations observed between FP archipelagos, Maoris and Hawaiians. PMID- 11044269 TI - Prevalence of point mutations in the dihydrofolate reductase and dihydropteroate synthetase genes of Plasmodium falciparum isolates from India and Thailand: a molecular epidemiologic study. AB - Pyrimethamine-sulfadoxine (PS) is used as a second-line treatment for P. falciparum malaria patients who fail to respond to chloroquine. Resistance to these drugs has been shown to encode with point mutations in dihydrofolate reductase (DHFR) and dihydropteroate synthetase (DHPS) genes. Our aim was to assess the comparative rate of point mutation occurring in DHFR and DHPS genes among P. falciparum isolates from India and Thailand where the use of PS is at a different rate. We used the mutation-specific polymerase chain reaction (PCR) technique and mutation-specific restriction digestion to determine the prevalence of DHFR and DHPS gene mutations at codons 16, 51, 59, 108, 164 and at 436, 437, 581 and 613, respectively. In the 89 clinical isolates from India, in the case of the DHFR gene, we found 71 of S108N, 10 of N51I, 28 of C59R and four of I164L types. Among the 50 isolates from Thailand the rate of point mutations in the DHFR gene was higher at four codon positions. We found 47 of S108N, 18 of N51I, 23 of C59R and 12 of I164L types. None of the isolates from either country possessed the paired mutations S108T and A16V. Mutations of the DHPS gene were less frequent among the Indian isolates: 4.5% showed DHPS gene mutation, two of S436F, A437G, A613T and two of S436F, A613T; whereas 66% (33/50) of the Thai isolates had mutated at codons 436, 437, 581 and 613 which include 13 of S436F, 15 of A437G, 19 of A581G and 25 of A613S/T, ranging from single to quadruple mutant types. Among the Indian isolates, DHFR point mutations were very frequent and 85/89 had a wild type DHPS genetic profile. The pattern of mutations in the samples from Thailand was different, as most were associated with point mutations in DHFR and DHPS genes. PMID- 11044270 TI - What is the effect of combining artesunate and praziquantel in the treatment of Schistosoma mansoni infections? AB - A group of 110 individuals with Schistosoma mansoni infection was investigated. Patients were allocated to one of three treatment groups and given artesunate or praziquantel alone or both in combination. Combined artesunate-praziquantel significantly increased the number of individuals cured at 5 weeks post treatment, but at 12 weeks was only better than artesunate alone and at 24 weeks there was no statistically significant difference between the three groups. Egg count reduction rate was similar to the rate obtained with praziquantel used alone. PMID- 11044271 TI - A century of research in tropical medicine in Hamburg: the early history and present state of the Bernhard Nocht Institute. PMID- 11044272 TI - The global programme to eliminate lymphatic filariasis. AB - Ten years ago, no one foresaw that in the year 2000 there would be a Global Programme to Eliminate Lymphatic Filariasis (GPELF) that is already 2 years old, active in 18 of the 80 endemic countries, and operating under a wholly new paradigm in public health - a paradigm affirming that public/private sector partnerships are essential in sharing both responsibilities and responses to global health problems. What has driven the LF Elimination Programme to this point? Where it is now headed? What will be required to sustain its momentum? What will its impact be? These are the issues addressed below. PMID- 11044273 TI - Household costs of 'malaria' morbidity: a study in Matale district, Sri Lanka. AB - Short-run economic consequences of 'malaria' on households were examined in a household survey in Matale, a malaria-endemic district of Sri Lanka. On average a household incurred a total cost of Rs 318 (US$ 7) per patient who fully recovered from 'malaria'. 24% of this was direct cost, 44% indirect cost for the patient and 32% indirect cost for the household. Direct costs were greater for those seeking treatment in the private sector. Notably a large proportion of direct costs was spent on complementary goods such as vitamins and foods considered to be nutritional. Indirect cost was measured and valued on the basis of output/ income losses incurred at the household level rather than using a general indicator such as average wage rate. Loss of output and wages accounted for the highest proportion of the indirect cost of the patients as well as the households. Relative to children, more young adults and middle-aged people had 'malaria' which also caused greater economic loss in these age groups. Women tended to care for patients rather than substitute their labour to cover productive work lost due to illness. We compare the methods used by other researchers for valuing indirect cost, demonstrating the significant impact that methods of measurement and valuation can have on the estimation of indirect cost, and justify the recommendation for methodological research in this area. PMID- 11044274 TI - Hyperreactive malaria in expatriates returning from sub-Saharan Africa. AB - The extreme presentation of hyperreactive malaria is hyperreactive malarial splenomegaly syndrome (HMS). Some patients present with a less pronounced syndrome. To investigate whether the degree of splenomegaly correlates with the degree of immune stimulation, whether prophylaxis or recent treatment play a role, and whether short therapy alone is effective, we examined retrospectively the medical records of expatriates with exposure to P. falciparum who attended our outpatient department from 1986 to 1997, particularly subacute symptoms or signs, strongly elevated malarial antibodies and elevated total serum IgM. We analysed duration of stay, prophlyaxis intake, spleen size, serum IgM levels and response to antimalarial treatment. Serum IgM levels were significantly higher in patients with larger splenomegaly. The use of chloroquine alone as treatment for presumptive or proved malaria attacks was correlated with larger spleen size. Short adequate antimalarial therapy resulted in marked improvement or complete recovery. In nine patients the hyperreactive response reappeared after re exposure, in four of them twice. We conclude that patients with subacute symptoms but without gross splenomegaly may have very high levels of IgM and malarial antibodies, and relapse on re-exposure, suggesting the existence of a variant of the hyperreactive malarial splenomegaly syndrome without gross splenomegaly. PMID- 11044275 TI - Chemoresistance of P. falciparum in urban areas of Yaounde, Cameroon. Part 1: Surveillance of in vitro and in vivo resistance of Plasmodium falciparum to chloroquine from 1994 to 1999 in Yaounde, Cameroon. AB - Chloroquine is indicated for the first-line treatment of uncomplicated malaria in most African countries. However, the spread of chloroquine-resistant Plasmodium falciparum requires periodic monitoring. Between 1994 and 1999, we studied the evolution of chloroquine resistance in adults (aged > 15 years) and children aged 5-15 years by using tests of therapeutic efficacy and in vitro assays. Responses to the 14-day in vivo test were classified according to the new criteria established by the World Health Organization. The results of the semi-microtest and the microtest were expressed as the 50% inhibitory concentration (IC50), and the threshold level of resistance was set at IC50 > 100 nM. The overall percentages of clinical and parasitological failures were 39.7% (31. 3% - 48.1%) and 48.8% (40.2% - 57.4%), respectively. Similarly, the percentage of isolates that were resistant in vitro was 52.5%. During the study, IC50 geometric mean varied between 84,6 nM and 149, 8 nM. The results of the in vitro assays agreed with those of tests of therapeutic efficacy (kappa coefficient = 0.69). The patients' chloroquine plasma levels were measured on day 0, day 3, day 7, and day 14. Drug measurement showed wide inter-individual variations and higher plasma levels in adults than in children. Some cases of therapeutic failure were associated with inadequate plasma levels of chloroquine. Our results confirm the high level of chloroquine resistance in Yaounde and suggest that the use of an alternative antimalarial drug for the first-line treatment of uncomplicated malaria is warranted. PMID- 11044276 TI - Chemoresistance of Plasmodium falciparum in the urban region of Yaounde, Cameroon. Part 2: Evaluation of the efficacy of amodiaquine and sulfadoxine pyrimethamine combination in the treatment of uncomplicated Plasmodium falciparum malaria in Yaounde, Cameroon. AB - The spread of chloroquine resistance or its stabilization at a high level calls for a change in the therapeutic strategy, including a possible replacement of chloroquine. We assessed and compared the efficacy of amodiaquine and sulfadoxine pyrimethamine in Yaounde. Of 140 adults and children > 5 years enrolled in the study, 59 in the amodiaquine and 58 in the sulfadoxine-pyrimethamine treatment group were followed until day 14. The efficacy of amodiaquine was 100%, whereas 12.1% of the patients treated with sulfadoxine-pyrimethamine responded with an early treatment failure. Side effects in both treatment groups were mild and did not require any specific treatment. We did in vitro drug assays for monodesethylamodiaquine (active metabolite of amodiaquine) and pyrimethamine and measured plasma levels of monodesethylamodiaquine, sulfadoxine, and pyrimethamine. Unlike amodiaquine, the results of the in vitro drug sensitivity test for pyrimethamine were not concordant with the clinical response. A wide inter-individual variation in the plasma drug levels was observed. Unlike chloroquine, the mean plasma concentrations did not vary with age. There was no significant difference in the plasma concentrations of sulfadoxine and pyrimethamine between patients responding with an adequate clinical response and those responding with treatment failure. Amodiaquine has several advantages over sulfadoxine-pyrimethamine combination and may be considered to be an effective drug in an endemic zone with a moderate level of chloroquine resistance. PMID- 11044277 TI - Season of birth is not associated with delayed childhood mortality in Upper River Division, The Gambia. AB - There is evidence that season of birth may predict adult mortality from infectious diseases in rural Gambia. Using data collected over a five-year period from the rural, eastern region of the Gambia, we examined whether the season of birth influences mortality in childhood. 26 894 births and 3776 deaths among children under the age of five years were recorded in this region during the period 1989-1993. The estimated 1-4 year population was 95 355. In children aged 1-4 years, the mortality rate per 1000 per year was 16.1 (95% CI 14.9, 17.2) for those born in the 'harvest' season (January to June), which was not significantly different from the rate of 17.9 (95% CI 16.7, 19.0) recorded for those born in the 'hungry' season (July to December) (age-stratified Mantel-Haenszel mortality ratio 0. 91, 95% CI 0.83, 1.01; p = 0.08). Nearly all deaths of 1-4 year olds were attributed to infectious diseases, with malaria accounting for over 40%. None of the cause-specific child mortality rates differed significantly according to the season of birth. These data suggest that beyond infancy, when it is easier to separate the effect of season on cause of death from that of the season of birth, there is no marked difference in the rate of death between Gambian children born in the harvest season and those born in the hungry season. PMID- 11044278 TI - Hepatitis B virus infection in Thai children. AB - We studied hepatitis B virus (HBV) transmission among 7416 Thai children from 148 schools in Kamphaeng Phet province, a rural part of northern Thailand. Their age ranged from 2 to 16 years (median 9 years). Between May 1991 and June 1992, 61 of 2593 (2.4%) in the cohort of susceptible children acquired anti-HBc immunoglobulin. Forty-seven of the 148 schools had children who acquired anti HBc. School seroconversion rates to anti-HBc varied from 0% to 23%. There was no correlation between percent of carriers in schools and percent of anti-HBc acquisition. Of the 61 children who acquired anti-HBc, eight (13%) became HBsAg carriers but only two were symptomatic, for a clinical to subclinical infection ration of 1 : 30. One of the two symptomatic children became an HBsAg carrier. Three (38%) of the eight who were persistently antigenemic developed antibody to hepatitis B virus e antigen. Males were 2.5 times (95% CI 1.4-4.3) more likely to acquire anti-HBc than females. Risk factors for acquisition of HBc in Thailand over a 9-month period were examined in a subset of 2412 susceptible children and later in a case-control study of 22 children who acquired anti-HBc and 59 age and sex-matched controls. Risks for acquiring anti-HBc were male gender and a history of bleeding gums. In comparing this study to an earlier pilot study among 9848 children from the same area in Thailand, the yearly antibody acquisition rate to anti-HBc among Thai children dropped from 5.7% in 1989 to 2.4% in 1992. A random sample of children in the pilot study showed that 16% were HBsAg positive and 27% had anti-HBc at the beginning of the study. 34% had markers for either anti-HBc or HBsAg. 12% were repeatedly positive for HBsAg a year later. PMID- 11044279 TI - Cost-effectiveness of management strategies for acute urethritis in the developing world. AB - OBJECTIVE: To recommend a cost-effective approach for the management of acute male urethritis in the developing world, based on the findings of a theoretical study. METHODS: A model was developed to assess the cost-effectiveness of three urethritis management strategies in a theoretical cohort of 1000 men with urethral syndrome. (1) All patients were treated with cefixime and doxycycline for gonococcal urethritis (GU) and nongonococcal urethritis (NGU), respectively, as recommended by WHO. (2) All patients were treated with doxycycline for NGU; treatment with cefixime was based on the result of direct microscopy of a urethral smear. (3) All patients were treated with cotrimoxazole or kanamycin for GU and doxycycline for NGU. Cefixime was kept for patients not responding to the first GU treatment. Strategy costs included consultations, laboratory diagnosis (where applicable) and drugs. The outcome was the rate of patients cured of urethritis. Cost-effectiveness was measured in terms of cost per cured urethritis. RESULTS: Strategy costs in our model depended largely on drug costs. The first strategy was confirmed as the most effective but also the most expensive approach. Cefixime should cost no more than US$ 1.5 for the strategy to be the most cost-effective. The second strategy saved money and drugs but proved a valuable alternative only when laboratory performance was optimal. The third strategy with cotrimoxazole was the least expensive but a low follow-up visit rate, poor treatment compliance or lower drug efficacy limited effectiveness. Maximizing compliance by replacing cotrimoxazole with single-dose kanamycin had the single greatest impact on the effectiveness of the third strategy. CONCLUSION: Our model suggested that a cost-effective approach would be to treat gonorrhoea with a single-dose antibiotic selected from locally available products that cost no more than US$ 1.5. PMID- 11044280 TI - Health seeking and perceived causes of tuberculosis among patients in Manila, Philippines. AB - Inefficient case finding is an important stumbling block to successful control of tuberculosis (TB). Multiple health seeking may account for delayed case finding. Health-seeking behaviour, health seeking delay, perceived causes, and perceived quality of care related to TB were studied in interviews with 319 sputum smear positive TB patients. The patients were treated in 22 governmental health centres of Malabon, a municipality of Metro Manila, Philippines. Only 29% of the respondents had gone first to a health centre after onset of TB-related symptoms, and more than half (53%) had initially consulted a private doctor. A chest X-ray was obtained for nearly everyone (97%). Two thirds of the patients (66%) had received a prescription for drugs, and 29% had purchased and taken anti-TB drugs for at least three weeks before they came to a governmental health centre. Concerning community interactions, 36% said they knew at least one person who had been treated for TB without success. The health seeking delay after symptom onset was relatively short - 64% of the respondents said they went to a health facility within 1 month. Case studies illustrate the rationale for health seeking and explain delayed initiation of appropriate treatment for many patients. Findings underscore the need for and indicate approaches to health communication for improved control of TB. Our findings from interview narratives also suggest that improved interpersonal skills of health centre staff and co-ordination between the private doctors and the health centres may substantially improve services for TB patients. PMID- 11044281 TI - Maternity care in rural Nepal: a health service analysis. AB - This study assesses the performance of maternity care and its specific service components (preventive interventions in antenatal care, antenatal screening, referral, obstetric care) in Banke District, Nepal, using a set of structure, process, and output/outcome indicators. Data sources included health service documents in 14 first level health units and two hospitals, covering 1378 pregnancies and 1323 deliveries, structured observations, antenatal exit interviews (n = 136) and interviews with maternity users (n = 146). Coverage of antenatal care (28%) and skilled delivery care (16%) was low. In antenatal care, preventive interventions were only partially implemented (effective iron supplementation in 17% of users). On average one minute was spent on individual counselling per consultation. 41% of pregnancies were identified as high risk and 15% received referral advice, which was followed in only 32%. Hospital deliveries accounted for 9.8% of all deliveries. Hospital-based maternal mortality was 6.8/1000 births and the stillbirth rate 70/1000. High rates of stillbirth were observed in breech delivery (258/1000 births), caesarean section (143/1000) and twin delivery (133/1000). The risk of stillbirth was higher for rural women (RR 2.3; 95% CI 1.51-3.50) and appeared to be related to low socio-economic status. Emergency admissions were rare and accounted for 3.4% of hospital deliveries or only 0.4% of all expected deliveries. There was hardly any accumulation of high risk pregnancies at hospital. The population-based rate of caesarean section was 1.1% (urban 2.3%, rural 0.2%). The estimated unmet obstetric need was high (82 cases or 61% of expected live-threatening maternal conditions did not receive appropriate intervention). The limited effectiveness of maternity care is the result of deficiencies of all service components. We propose a two-pronged approach by starting quality improvement of maternity care from both ends of maternity services: preventive interventions for all women and hospital-based obstetric care. Antenatal screening needs to be rationalized by reducing inflated risk catalogues that result in stereotypical and often rejected referral advice. PMID- 11044282 TI - Viewpoint: stability of insulin in tropical countries. PMID- 11044283 TI - Are there simple measures to reduce the risk of HIV infection through blood transfusion in a Zambian district hospital? AB - OBJECTIVE: To quantify the potential impact of simple measures to reduce the risk of iatrogenic HIV infection through blood transfusion in a Zambian district hospital. METHODS: Three studies were conducted at St. Francis' Hospital, Katete, Zambia: (1) From 1991 to 1995 HIV seroprevalence among all listed blood donors and the impact of proper subgroup selection were studied retrospectively; (2) the sensitivity of locally used rapid antibody assays (HIV-spot/Wellcozyme HIV 1 & 2) for the detection of HIV in donor blood and the influence of the expiration date of the tests on this sensitivity were determined prospectively from June 1993 until March 1994 by screening all consecutive surgical patients and blood donors; (3) the number of unnecessary blood transfusions was determined retrospectively from January 1995 through January 1996 and prospectively from February 1996 through March 1996, and possibilities to reduce the total number of blood transfusions were considered. RESULTS: (1) Excluding prisoners, who have an HIV seroprevalence of 19-25%, from the donor population significantly reduces the overall HIV seroprevalence from 13-16% to 8-9% (P < 0. 01). (2) Under local circumstances the sensitivity of the used rapid antibody assays was 6.8-17.9% lower than claimed by the manufacturer. Usage of non-expired tests increased the sensitivity significantly from 88.2% to 91.7% (P < 0.05). (3) None of the 294 studied blood transfusions can be classified as inappropriate according to international standards. CONCLUSIONS: Simple measures such as proper subgroup selection among blood donors and correct use of non-expired tests may decrease the risk of iatrogenic HIV transmission. Stricter indications for blood transfusions will not substantially reduce the number of transfusions. PMID- 11044284 TI - Dermatophytosis consensus meeting: Introduction. PMID- 11044285 TI - Overview: The treatment of dermatophytosis. PMID- 11044286 TI - The treatment of dermatophytosis: safety considerations. PMID- 11044287 TI - Dermatophytosis: epidemiological and microbiological update. PMID- 11044288 TI - Evaluation of in vitro susceptibility of dermatophytes to oral antifungal agents. PMID- 11044289 TI - Onychomycosis and tinea pedis in patients with diabetes. PMID- 11044290 TI - Dermatophyte infections in human immune deficiency virus (HIV) disease. PMID- 11044291 TI - Overstating the ordinary. PMID- 11044293 TI - 2010 nursing PMID- 11044292 TI - Moral distress in everyday ethics. PMID- 11044294 TI - A conversation with Robert Blendon about public opinion and health care, nursing and the 2000 presidential election. Interview by Peter I. Buerhaus. PMID- 11044295 TI - Retirement, the nursing workforce, and the year 2005. AB - This analysis of workforce projections confirms that early employment withdrawal by registered nurse baby boomers could have a profound effect on US health care. The available policy mechanisms to encourage or discourage any early withdrawal require several years to implement, which makes timely decisions imperative. PMID- 11044297 TI - Secondary data analysis: new perspective for adolescent research. AB - Adolescents are at high risk for preventable health problems, but realities of research funding and obtaining data from large samples make it difficult for many researchers to answer important questions. Secondary analysis of existing data sets is described as a reasonable alternative in spite of its limitations. Methods and available resources are identified along with suggestions for research priorities with adolescents. PMID- 11044296 TI - Accelerated second degree advanced practice nurses: how do they fare in the job market? AB - Accelerated, nontraditional, advanced practice nursing programs are an alternative way to increase the supply of nurse practitioners. This study profiles demographic and job characteristics of second degree, non-nurse college graduates who pursued graduate degrees in nursing. Graduates' sex, age, income, previous education, nursing experience, factors describing the scope of the advanced practice role, and quality of the educational experience were studied. Data were collected from 29 graduates (57%) from Virginia Commonwealth University's accelerated second-degree nursing program from 1995 through 1999. The findings have implications for nursing educators, health care administrators, employers, and other persons who plan and recruit for this type of nursing education program. PMID- 11044298 TI - Recruiting families of color from the inner city: insights from the recruiters. AB - Family interviewers hired to recruit families of color from low-income communities into a health promotion/prevention research project participated in a focus group to describe their experiences. The family interviewers described (1) recruitment techniques, (2) personal attributes they thought made a difference in their recruitment and retention efforts, (3) barriers they faced during recruitment, and (4) the experience of working for a white researcher. PMID- 11044300 TI - A national voice for nursing research: council for the advancement of nursing science PMID- 11044301 TI - Bringing oncology nursing front and center. PMID- 11044299 TI - Challenges and opportunities in psychiatric-mental health nursing. PMID- 11044303 TI - Expert panel on primary care research networks PMID- 11044302 TI - Policy leadership training institute-another successful year PMID- 11044304 TI - Expert panels: what are they? PMID- 11044305 TI - Re: undereducated, aging and ... a cycle of decline? PMID- 11044306 TI - On the future of nursing. PMID- 11044307 TI - To the editor PMID- 11044308 TI - The innovation imperative. PMID- 11044309 TI - Midterm follow-up of patients who underwent removal of a left ventricular assist device after cardiac recovery from end-stage dilated cardiomyopathy. AB - OBJECTIVE: Cardiac recovery in end-stage idiopathic dilated cardiomyopathy recently occurred after temporary support with a left ventricular assist device. We report the case histories of patients who underwent removal of the device more than 4 years ago. METHODS: Since June 1994, 23 patients with end-stage idiopathic dilated cardiomyopathy who were supported by a left ventricular assist device or biventricular assist device for 1 to 26 months (mean, 6 months) underwent removal of the device after complete or extensive cardiac recovery, as revealed by echocardiography. RESULTS: Seven patients (group A) had recurrent cardiac failure after 4 to 24 months. Transplantation was performed in 6 patients, and one died while on the waiting list. Three patients died of noncardiac causes within a period of 4 months and 3 days after removal of the assist device. Stable cardiac recovery occurred in 13 patients (group B) for 3 to 49 months (mean, 23 months). At the time of implantation, there were no significant differences between the groups with regard to age, hemodynamics, left ventricular ejection fraction, left ventricular internal diameter in diastole, and autoantibody levels. The increase of ejection fraction and the decrease of left ventricular internal diameter in diastole after 2 months were highly significant. The patients in group A had longer histories of heart failure and first cardiac symptoms and duration of assist when compared with group B. Group B demonstrated a quicker cardiac recovery on the assist device, and thus support was shorter. Also, the degree of recovery at assist device explantation was more complete in group B. The age at the time of device placement was the only influencing factor for duration on the assist device. The probability of recurrence of heart failure was influenced by the duration of heart failure. CONCLUSIONS: In selected patients with idiopathic dilated cardiomyopathy, lasting recovery can be achieved after unloading with a left ventricular assist device. Lasting cardiac recovery seems to be related to functional normalization and a more rapid recovery during the unloading period. PMID- 11044310 TI - Assisted venous drainage presents the risk of undetected air microembolism. AB - OBJECTIVES: The proliferation of minimally invasive cardiac surgery has increased dependence on augmented venous return techniques for cardiopulmonary bypass. Such augmented techniques have the potential to introduce venous air emboli, which can pass to the patient. We examined the potential for the transmission of air emboli with different augmented venous return techniques. METHODS: In vitro bypass systems with augmented venous drainage were created with either kinetically augmented or vacuum-augmented venous return. Roller or centrifugal pumps were used for arterial perfusion in combination with a hollow fiber oxygenator and a 40-micrometer arterial filter. Air was introduced into the venous line via an open 25-gauge needle. Test conditions involved varying the amount of negative venous pressure, the augmented venous return technique, and the arterial pump type. Measurements were recorded at the following sites: pre-arterial pump, post arterial pump, post-oxygenator, and patient side. RESULTS: Kinetically augmented venous return quickly filled the centrifugal venous pump with macrobubbles requiring continuous manual clearing; a steady state to test for air embolism could not be achieved. Vacuum-augmented venous return handled the air leakage satisfactorily and microbubbles per minute were measured. Higher vacuum pressures resulted in delivery of significantly more microbubbles to the "patient" (P <.001). The use of an arterial centrifugal pump was associated with fewer microbubbles (P =.02). CONCLUSIONS: Some augmented venous return configurations permit a significant quantity of microbubbles to reach the patient despite filtration. A centrifugal pump has air-handling disadvantages when used for kinetic venous drainage, but when used as an arterial pump in combination with vacuum-assisted venous drainage it aids in clearing air emboli. PMID- 11044311 TI - Temporal endothelin dynamics of the myocardial interstitium and systemic circulation in cardiopulmonary bypass. AB - OBJECTIVE: Increased systemic levels of the bioactive peptide endothelin 1 during and after cardioplegic arrest and cardiopulmonary bypass have been well documented. However, endothelin 1 is synthesized locally, and therefore myocardial endothelin 1 production during and after cardiopulmonary bypass remains unknown. METHODS: Pigs (n = 11) were instrumented for cardiopulmonary bypass, and cardioplegic arrest was initiated. Myocardial interstitial and systemic arterial levels of endothelin 1 were measured before cardiopulmonary bypass, throughout bypass and cardioplegic arrest (90 minutes), and up to 90 minutes after separation from bypass. Myocardial interstitial endothelin 1 was determined by microdialysis and radioimmunoassay. RESULTS: Baseline myocardial endothelin 1 levels were higher than systemic endothelin 1 levels (25.6 +/- 6.7 vs 8.3 +/- 1.1 fmol/mL, P <.05). With the onset of bypass, myocardial endothelin 1 increased by 327% +/- 92% from baseline (P <.05), which preceded the increase in systemic endothelin 1 levels. CONCLUSION: Myocardial compartmentalization of endothelin 1 exists in vivo. Cardiopulmonary bypass and cardioplegic arrest induce temporal differences in endothelin 1 levels within the myocardial interstitium and systemic circulation, which, in turn, may influence left ventricular function in the postbypass period. PMID- 11044312 TI - Aortic reconstruction in hypoplastic left heart syndrome-A reappraisal. PMID- 11044313 TI - Modified Norwood procedure with a high-flow cardiopulmonary bypass strategy results in low mortality without late arch obstruction. AB - OBJECTIVE: The results of our modification of the stage I Norwood procedure, in which we use only autologous tissue to reconstruct the aortic arch, were reviewed. A high-flow, low-pressure cardiopulmonary bypass protocol (with phenoxybenzamine), before and after a period of deep hypothermic circulatory arrest, was used. METHODS: Between 1993 and 1999, 59 patients, aged 1 to 353 days (median 4 days) and weighing 1.7 to 6.8 kg (median 3.2 kg), underwent a modified Norwood procedure. The ascending aortic diameter ranged from 1.5 to 8 mm (median 3 mm). The modified Blalock-Taussig shunt was 3 mm in 21 patients (36%) and 3.5 mm or larger in 38 patients (64%). RESULTS: Deep hypothermic circulatory arrest and cardiopulmonary bypass times ranged from 15 to 64 minutes (median 37 minutes) and 44 to 144 minutes (median 88 minutes), respectively. Early postoperative survival was 83%. By univariate analysis, early mortality was associated with an ascending aortic diameter of 2.5 mm or less (P =.01). Weight, circulatory arrest and bypass times, diagnosis (hypoplastic left heart syndrome vs variant), shunt size, and date of the procedure did not affect survival. For a median follow-up period of 37 months (range 4-63 months), 42 (61%) patients underwent bidirectional cavopulmonary shunts, 10 (17%) had Fontan operations, and 1 patient underwent transplantation after a bidirectional cavopulmonary shunt. Eight patients subsequently died, for a 1-year actuarial survival of 72% (95% confidence interval: 60%-84%). Neoaortic arch obstruction was corrected in 3 patients (5%). CONCLUSIONS: At intermediate-term follow-up, our modification of the Norwood procedure together with our perioperative strategies has resulted in acceptable outcomes with a low incidence of neoaortic arch obstruction. Patients with a small ascending aortic diameter have emerged as a high-risk group, but a recent technical modification may improve the outlook for these patients. PMID- 11044314 TI - Extra-anatomic aortic bypass via sternotomy for complex aortic arch stenosis in children. AB - OBJECTIVE: Recurrent aortic narrowing after repair of aortic coarctation or interrupted aortic arch, as well as diffuse, long-segment aortic hypoplasia, can be difficult to manage. Extra-anatomic ascending aorta-descending aorta bypass grafting through a sternotomy is an alternative approach for this problem. METHODS: Since 1985, 19 patients aged 2 months to 18 years (mean 10.7 years) underwent extra-anatomic bypass with 10- to 30-mm Dacron grafts. The initial diagnosis was coarctation with hypoplastic arch in 15, interrupted aortic arch in 3, and diffuse long-segment aortic hypoplasia in 1. Seventeen of the children had a total of 22 previous operations: transthoracic interposition or bypass graft (n = 7), end-to-end anastomosis (n = 7), subclavian arterioplasty (n = 6), and synthetic patch (n = 2). The mean time from initial repair was 8.0 years (range 0.6-18 years). Three children had previous sternotomies. Cardiopulmonary bypass was avoided in all but 6 patients (5 with simultaneous intracardiac repairs). RESULTS: No hospital or late deaths occurred. On follow-up from 4 months to 14.7 years (mean 7.9 years), no reoperations for recurrent stenosis were performed. Two patients have arm-to-leg pressure gradients: 20 mm Hg at rest in 1 patient and a 60-mm Hg systolic exercise gradient with no resting gradient in the other. One patient required exclusion of an aortic aneurysm at the old coarctation repair site 13 years after extra-anatomic bypass. Three children had subsequent successful cardiac operations. CONCLUSIONS: Extra-anatomic bypass is an effective and relatively easy approach for selected cases of complex or reoperative aortic arch obstruction. It should be considered as an alternative operative technique for complex aortic arch reconstruction. PMID- 11044316 TI - Pulmonary microvessel density is a marker of angiogenesis in children after cavopulmonary anastomosis. AB - OBJECTIVE: Pulmonary arteriovenous malformations cause progressive cyanosis in children after cavopulmonary anastomosis and may be due to abnormal angiogenesis. We determined the microvessel density, a marker of angiogenesis, in the lungs of children after cavopulmonary anastomosis. METHODS: Lung biopsy specimens were obtained from 8 children after cavopulmonary anastomosis and from 4 control patients. Three of the 8 children undergoing cavopulmonary anastomosis had clinical and angiographic evidence of pulmonary arteriovenous malformations, whereas the other 5 were free of symptoms. Routine histologic and immunohistologic stains were performed with a primary antibody to von Willebrand factor. Microvessel staining for von Willebrand factor was determined for 10 fields (200x) per patient. RESULTS: Patients with and without pulmonary arteriovenous malformations after cavopulmonary anastomosis demonstrated significantly increased microvessel density compared with control subjects (32.7 +/- 2.8 vs 9.3 +/- 4.6, P =.02, and 31.5 +/- 15.7 vs 9.3 +/- 4.6, P =.01, respectively). There was no difference in microvessel density in children with and without clinically apparent pulmonary arteriovenous malformations after cavopulmonary anastomosis (P =.9). The children with pulmonary arteriovenous malformations had numerous greatly dilated vessels that were absent in the asymptomatic children after cavopulmonary anastomosis. CONCLUSIONS: After cavopulmonary anastomosis, pulmonary microvessel density is increased even in the absence of clinically apparent pulmonary arteriovenous malformations, supporting the presence of a constant angiogenic stimulus. Children with clinically apparent pulmonary arteriovenous malformations possess large numbers of greatly dilated pulmonary microvessels, which are absent in asymptomatic children after cavopulmonary anastomosis. These results suggest that the transition to clinically apparent pulmonary arteriovenous malformations may be due to mechanisms that lead to vessel dilation and remodeling. PMID- 11044315 TI - Modifications to the cavopulmonary anastomosis do not eliminate early sinus node dysfunction. AB - OBJECTIVE: To determine whether operations that theoretically jeopardize the sinus node (hemi-Fontan and/or lateral tunnel Fontan procedures) are associated with a greater risk of sinus node dysfunction than those that theoretically spare the sinus node (bidirectional Glenn and/or extracardiac conduit). METHODS: Between January 1, 1996, and December 31, 1999, a prospective cohort study was conducted evaluating the incidence of sinus node dysfunction in patients undergoing a bidirectional Glenn or hemi-Fontan procedure and those in whom the Fontan repair was completed with either an extracardiac conduit or a lateral tunnel. Sinus node dysfunction was defined (1) as a heart rate more than 2 SD below age-adjusted norms or (2) as a predominant junctional rhythm and/or a sinus pause of more than 3 seconds as determined by the resting electrocardiogram and/or ambulatory monitoring at hospital discharge. RESULTS: Fifty-one patients had a bidirectional Glenn shunt (mean age 7.8 +/- 5.1 months) and 79 a hemi Fontan procedure (mean age 6.9 +/- 2.8 months). The incidence of sinus node dysfunction on postoperative day 1 was significantly higher after the hemi-Fontan (36%) than after the bidirectional Glenn shunt (9.8%); however, by hospital discharge this difference was no longer apparent (hemi-Fontan [8%]; bidirectional Glenn [6%]; P = not significant). No difference in early sinus node dysfunction was discernible after the extracardiac conduit (4/30 [13%]) compared with the lateral tunnel Fontan procedure (6/46 [13%]) (P = not significant). No diagnostic or perioperative variables were predictive of sinus node dysfunction. CONCLUSIONS: Avoidance of surgery near the sinus node has no discernible effect on the development of early sinus node dysfunction. Thus, concerns about early sinus node dysfunction should not override patient anatomy or surgeon preference as determinants of which cavopulmonary anastomosis to perform. PMID- 11044317 TI - Perioperative complications after living donor lobectomy. AB - OBJECTIVE: Clinical lung transplantation has been limited by availability of suitable cadaveric donor lungs. Living donor lobectomy provides right and left lower lobes from a pair of living donors for each recipient. We reviewed our experience with living donor lobectomy from July 1994 to February 2000. METHODS: Sixty-two donor lobectomies were performed. The hospital and outpatient records of these 62 donors were retrospectively analyzed to examine the incidence of perioperative complications. RESULTS: Twenty-four (38.7%) of 62 donors had no perioperative complications and had a median length of hospital stay of 5.0 days. Thirty-eight (61.3%) of 62 donors had postoperative complications. Twelve major complications occurred in 10 patients and included pleural effusions necessitating drainage (n = 4), bronchial stump fistulas (n = 3), bilobectomy (n = 1), hemorrhage necessitating red cell transfusion (n = 1), phrenic nerve injury (n = 1), atrial flutter ultimately necessitating electrophysiologic ablation (n = 1), and bronchial stricture necessitating dilatation (n = 1). These 38 donors had 55 minor complications including persistent air leaks (n = 9), pericarditis (n = 9), pneumonia (n = 8), arrhythmia (n = 7), transient hypotension necessitating fluid resuscitation (n = 4), atelectasis (n = 3), ileus (n = 3), subcutaneous emphysema (n = 3), urinary tract infections (n = 2), loculated pleural effusions (n = 2), transfusion (n = 2), Clostridium difficile colitis (n = 1), puncture of a saline breast implant (n = 1), and severe contact dermatitis secondary to adhesive tape (n = 1). There were no postoperative deaths and only 1 donor required surgical re-exploration. CONCLUSIONS: Living donor lobectomy can be performed with low mortality and remains an important alternative for potential recipients unable to wait for cadaveric lung allografts. However, morbidity is high and must be considered when potential living donors are being counseled. PMID- 11044318 TI - Epidermal growth factor augments postpneumonectomy lung growth. AB - OBJECTIVE: Epidermal growth factor has been shown to play an important role in prenatal and postnatal lung development, but little is known about its effects on adult lung growth. We hypothesized that postpneumonectomy compensatory lung growth can be augmented by the administration of epidermal growth factor. METHODS: Adult Sprague-Dawley rats were divided into 3 groups. Sham left thoracotomy was performed in the first group (group C), left pneumonectomy in the second group (group P), and left pneumonectomy with administration of epidermal growth factor (0.2 microgram/g body weight intraperitoneally, at 72-hour intervals) in the third group (group E). The right lung growth was studied in each group 1, 3, 5, 10, and 21 days after the operation. Lung weights (in grams) and volumes (in milliliters) were expressed as a ratio to the total body weight (in kilograms) (lung weight and volume indices). Epidermal growth factor receptor was quantitated by using Western blotting. RESULTS: Using analysis of variance and contrast analysis, we noted a significant increase in lung weight index in group E versus group P rats at 3 days (3.08 vs 2.75; P =.034) and 21 days (4.62 vs 3.61; P =.006). Lung volume index was significantly increased in group E versus group P rats at 5 (16.98 vs 15.09), 10 (24.48 vs 18.81), and 21 (28.54 vs 21.01) days (P <.001). Epidermal growth factor receptor was noted to be up regulated in the lungs of animals that received exogenous epidermal growth factor. CONCLUSIONS: This study demonstrates that administration of exogenous epidermal growth factor has a significant effect on postpneumonectomy lung growth. This process may be mediated by an up-regulation of growth factor receptor expression in the contralateral lung. PMID- 11044319 TI - A novel approach with magnetic resonance imaging used for the detection of lung allograft rejection. AB - OBJECTIVE: Although various techniques have been explored for the detection and quantification of allograft transplant rejection, a practical and reliable method that is noninvasive is still elusive. METHODS: For our magnetic resonance imaging experiments, we have developed a new rat model of heterotopic lung transplantation to the inguinal region. Allogeneic transplants (DA to Brown Norway) were performed with and without cyclosporine A (INN: ciclosporin) treatment, with syngeneic transplants (Brown Norway to Brown Norway) serving as controls (n = 6 per group). Magnetic resonance images were obtained with a gradient echo method before and after injection of ultra-small superparamagnetic iron oxide particles. RESULTS: At day 5, allogeneic transplants without cyclosporine A treatment showed a grade 4 rejection histologically. A significantly lower magnetic resonance signal was seen 24 hours after injection of ultra-small superparamagnetic iron oxide particles compared with the preinjection image (346 +/- 7.6 vs 839 +/- 43.4 arbitrary units; P <. 05). Syngeneic transplants showed no evidence of rejection histologically and no differences in magnetic resonance imaging signals between the images before and after injection of ultra-small superparamagnetic iron oxide particles (863 +/- 18.8 vs 880 +/- 22.5). Allotransplants treated with cyclosporine A showed a grade 2 rejection histologically. The change in magnetic resonance signals in that group was small but showed a significant decrease in signal intensity after injection (646 +/- 10.5 vs 889 +/- 23.5, P <.05). Immunohistochemistry and iron staining of the allografts indicated that ultra-small superparamagnetic iron oxide particles were taken up by the infiltrating macrophages that accumulated at the rejection site. CONCLUSIONS: We have demonstrated a novel approach for the detection of acute lung allograft rejection using magnetic resonance imaging coupled with injection of ultra-small superparamagnetic iron oxide particles. Despite its limitations, our method might be a first step toward a potential clinical application. PMID- 11044320 TI - Timed barium esophagogram: A simple physiologic assessment for achalasia. AB - OBJECTIVE: Success of achalasia therapy is difficult to determine because repeated physiologic study is impractical and symptoms are subjective. Timed barium esophagography directly measures esophageal emptying and is simple to perform. This study (1) evaluates the assessment of myotomy by timed barium esophagography and (2) compares it with premyotomy and postmyotomy symptoms. METHODS: Fifty patients ingested 250 mL low-density barium and had upright films at 1, 2, and 5 minutes premyotomy. Forty-five underwent repeat timed barium esophagography 8 weeks (median) postmyotomy. Premyotomy and postmyotomy height and width of the barium column were compared and related to symptoms. RESULTS: At 1, 2, and 5 minutes premyotomy, median barium column height was 19, 17, and 15 cm, and width was 5.2, 4.8, and 4.5 cm, respectively. Surgery reduced these to 7.0, 5.0, and 1.0 cm and to 3.5, 3.0, and 1.0 cm, respectively (P <.001). Postmyotomy complete esophageal emptying was seen in 29%, 36%, and 49% at 1, 2, and 5 minutes. Postmyotomy height was unrelated (r approximately 0.2) to premyotomy height but was directly related to premyotomy width (r = 0.3-0.5; P <.05); postmyotomy width was directly related to premyotomy width (r approximately 0.6; P <.001). Premyotomy dysphagia was more severe when little change in width occurred from 1 to 5 minutes (r = 0.26, P =.07). Premyotomy regurgitation was more severe the higher the barium column (r approximately 0.4, P <.007). Surgery relieved symptoms in the majority of patients (grade 2-5 dysphagia from 72% to 4%, grade 2-5 regurgitation from 79% to 4%). Postmyotomy symptoms were unrelated to the timed barium esophagogram. CONCLUSIONS: (1) The timed barium esophagogram gives objective confirmation of successful myotomy. (2) Symptoms are unreliable in assessing esophageal emptying. PMID- 11044321 TI - Adenovirus-mediated gene transfer of human interleukin 10 ameliorates reperfusion injury of rat lung isografts. AB - OBJECTIVE: The objective of this study was to examine the feasibility of human interleukin 10 gene transfer into rat lung isografts and to investigate the effect of gene transfer on subsequent ischemia-reperfusion injury. METHODS: Male F344 rats were divided into 4 groups and underwent left lung isotransplantation. Twenty-four hours before harvest, 5 x 10E9 pfu (group I, n = 6) or 1 x 10E10 pfu (group II, n = 7) of AdRSVhIL-10 was intravenously administered to donor rats. In group I-C (n = 6) and group II-C (n = 6), serving as controls, 5 x 10E9 pfu and 1 x 10E10 pfu of AdCMVLacZ were administered, respectively. Grafts were preserved for 18 hours at 4 degrees C before implantation and assessed 24 hours after reperfusion. Transgene expression of human interleukin 10 was assessed by both reverse transcriptase-polymerase chain reaction and immunohistochemistry. Graft inducible nitric oxide synthase, tumor necrosis factor alpha, intercellular adhesion molecule-1, growth-regulated gene product/cytokine-induced neutrophil chemoattractant-1, and monocyte chemotactic protein-1 mRNA expression were assessed by reverse transcriptase-polymerase chain reaction. Isograft gas exchange, exhaled nitric oxide, and myeloperoxidase activity were also analyzed. RESULTS: Dose-dependent transgene expression was detected by reverse transcriptase-polymerase chain reaction and immunohistochemistry. Arterial PO (2) in groups I (164.72 +/- 85.3 mm Hg) and II (153.19 +/- 113 mm Hg) was significantly higher than in groups I-C (82.37 +/- 19.1 mm Hg) and II-C (77.95 +/ 33.4 mm Hg) (P =.022 and P =.031, respectively). Arterial PCO (2) in group I (33.40 +/- 6.80 mm Hg) was significantly lower than in group I-C (51.23 +/- 11.9 mm Hg) (P =.0096). Myeloperoxidase activity in group II (0.083 +/- 0.031 DeltaOD. min(-1). mg(-1)) was significantly lower than in group II-C (0.117 +/- 0.028 DeltaOD. min(-1). mg(-1)) (P =.044). The inducible nitric oxide synthase mRNA expression in group II (0.627 +/- 0.28) was significantly lower than in group II C (1.125 +/- 0.63) (P =. 039). CONCLUSION: Adenovirus-mediated human interleukin 10 gene transfer in vivo into lung isografts ameliorates subsequent ischemia reperfusion injury. This results in improved graft gas exchange, reduced neutrophil sequestration, and down-regulation of graft inducible nitric oxide synthase mRNA expression. PMID- 11044322 TI - The atherosclerotic aorta at aortic valve replacement: surgical strategies and results. AB - BACKGROUND: Aortic valve replacement in patients with severe atherosclerosis of the ascending aorta poses technical challenges. The purpose of this study was to examine operative strategies and results of aortic valve replacement in patients with a severely atherosclerotic ascending aorta that could not be safely crossclamped. PATIENTS AND METHODS: From January 1990 to December 1998, 4983 patients had aortic valve surgery; of these, 62 (1.2%) patients had a severely atherosclerotic ascending aorta and required hypothermic circulatory arrest to facilitate aortic valve replacement. They form the study group. RESULTS: All patients had hypothermic circulatory arrest, but several different strategies were used to manage the ascending aorta. These techniques included aortic valve replacement with the use of hypothermic circulatory arrest (39%), ascending aortic endarterectomy (26%), ascending aortic replacement (19%), aortic inspection and crossclamping during hypothermic circulatory arrest (10%), and balloon occlusion of the ascending aorta (6%). Duration of hypothermic circulatory arrest was substantially longer for patients having aortic valve replacement with hypothermic circulatory arrest than for all other strategies. Hospital mortality was 14%, and 10% of patients had strokes. Increasing New York Heart Association functional class and impaired left ventricular function were risk factors for hospital mortality. Choice of operative technique did not influence patient outcome; however, no patient who underwent replacement of the ascending aorta had a stroke. CONCLUSIONS: Aortic valve replacement in patients with severe atherosclerosis of the ascending aorta is associated with increased operative morbidity and mortality. Complete aortic valve replacement during hypothermic circulatory arrest, the "no-touch" technique, requires a prolonged period of circulatory arrest. Ascending aortic replacement is a preferred technique, as it requires a short period of hypothermic circulatory arrest and results in comparable mortality with a low risk of stroke. PMID- 11044323 TI - The effects of ring annuloplasty on mitral leaflet geometry during acute left ventricular ischemia. AB - BACKGROUND: The perturbed mitral leaflet geometry that leads to acute ischemic mitral regurgitation during acute left ventricular ischemia has not been quantified, nor is it known whether annuloplasty rings affect these detrimental changes in leaflet geometry. METHODS: Radiopaque markers were implanted on both mitral leaflets and around the anulus in 3 groups of sheep: one group without rings served as the control group (n = 7); the others underwent Duran (n = 6; Medtronic Heart Valve Division, Minneapolis, Minn) or Carpentier-Edwards Physio (n = 5; Baxter Cardiovascular Division, Santa Ana, Calif) ring annuloplasty. After recovery, 3-dimensional marker coordinates were obtained by means of biplane videofluoroscopy before and during acute posterolateral left ventricular ischemia. Leaflet geometry was defined by measuring distances between annular and leaflet markers and perpendicular distances to the leaflet markers from a best fit annular plane. RESULTS: In all control animals, left ventricular ischemia was associated with acute ischemic mitral regurgitation and apical displacement (away from the annular plane) of the posterior leaflet edge and base markers by 0.6 +/- 0.4 mm (P =.01) and 0.7 +/- 0.2 mm (P <.001), respectively. The distance between the posterior leaflet markers and the mid-posterior anulus did not change significantly during ischemia. The anterior leaflet edge marker extended 1.0 +/- 0. 5 mm (P =.01) away from the mid-anterior anulus during ischemia, but compared with its nonischemic position, the anterior leaflet was not displaced apically away from the annular plane. In all animals in the Duran and Physio groups, leaflet geometry was unchanged during ischemia, and acute ischemic mitral regurgitation was not detected. CONCLUSION: Acute ischemic mitral regurgitation was associated with restricted motion of the posterior leaflet and extension of the anterior leaflet. Annuloplasty rings prevented these geometric perturbations of the mitral leaflets during acute left ventricular ischemia and preserved valvular competence. PMID- 11044324 TI - In health care, geography is destiny. PMID- 11044325 TI - Selection of a cardiac surgery provider in the managed care era. AB - OBJECTIVE: Many health planners promote the use of competition to contain cost and improve quality of care. Using a standard econometric model, we examined the evidence for "value-based" cardiac surgery provider selection in eastern Massachusetts, where there is significant competition and managed care penetration. METHODS: McFadden's conditional logit model was used to study cardiac surgery provider selection among 6952 patients and eight metropolitan Boston hospitals in 1997. Hospital predictor variables included beds, cardiac surgery case volume, objective clinical and financial performance, reputation (percent out-of-state referrals, cardiac residency program), distance from patient's home to hospital, and historical referral patterns. Subgroup analyses were performed for each major payer category. RESULTS: Distance from patient's home to hospital (odds ratio 0.90; P =.000) and the historical referral pattern from each patient's hometown (z = 45.305; P =.000) were important predictors in all models. A cardiac surgery residency enhanced the probability of selection (odds ratio 5.25; P =.000), as did percent out-of-state referrals (odds ratio 1.10; P =.001). Higher mortality rates were associated with decreased probability of selection (odds ratio 0.51; P =.027), but higher length of stay was paradoxically associated with greater probability (odds ratio 1.72; P =.000). Total hospital costs were irrelevant (odds ratio 1.00; P =.179). When analyzed by payer subgroup, Medicare patients appeared to select hospitals with both low mortality (odds ratio 0.43; P =.176) and short length of stay (odds ratio 0.76; P =.213), although the results did not achieve statistical significance. The commercial managed care subgroup exhibited the least "value-based" behavior. The odds ratio for length of stay was the highest of any group (odds ratio = 2.589; P =.000) and there was a subset of hospitals for which higher mortality was actually associated with greater likelihood of selection. CONCLUSIONS: The observable determinants of cardiac surgery provider selection are related to hospital reputation, historical referral patterns, and patient proximity, not objective clinical or cost performance. The paradoxic behavior of commercial managed care probably results from unobserved choice factors that are not primarily based on objective provider performance. PMID- 11044326 TI - Bilateral internal thoracic artery grafting: long-term clinical and angiographic results of in situ versus Y grafts. AB - BACKGROUND: We evaluated whether bilateral internal thoracic arteries provide the same long-term results when used as in situ grafts and as Y grafts. METHODS AND RESULTS: From October 1991 to February 2000, 1818 patients had bilateral internal thoracic arteries used as in situ (n = 1378, group A) or as Y grafts (n = 440, group B). The number of anastomoses per patient and the number of bilateral internal thoracic artery anastomoses per patient were higher in group B (3.1 +/- 0.9 and 2.7 +/- 0.9) than in group A (2.9 +/- 0.8 and 2.2 +/- 0.6) (both P <.001). The number of right internal thoracic artery anastomoses per patient rose from 1.0 +/- 0. 3 in group A to 1.4 +/- 0.6 in group B (P <.001), and the number of sequential anastomoses per right internal thoracic artery graft rose from 4.1% to 34.3% (P <.001). Thirty-day mortality was 2.0% in group A versus 2.5% in group B (P = not significant). No difference in postoperative course was detected. Eight-year survivals were 95.8% +/- 2.7% in group A versus 94.8% +/- 4.0% in group B (P = not significant), and event-free survivals were 95.2% +/- 2.9% in group A versus 93.6% +/- 4.4% in group B (P = not significant). Early angiograms were obtained in 295 patients (945 anastomoses, 863 distal and 82 proximal Y grafts), 213 patients (611) in group A and 82 patients (334) in group B. Patency rate was 98.8% in group A and 96.0% in group B (P = not significant), whereas grade A patency rate was 97.2% in group A and 96.4% in group B (P = not significant). Late angiograms were obtained in 88 patients (25 in group A and 63 in group B) at a mean of 17.5 +/- 18.4 months: patency rate was 100% in group A and 99.2 in group B (P = not significant), and grade A patency rate was 98.6% in group A and 98.8% in group B (P = not significant). No Y anastomosis was occluded or stenosed. COMMENT: Survival, incidence of cardiac events, and angiographic patency in the early and late phases are similar for bilateral internal thoracic arteries used either in situ or as Y grafts. However, Y grafting with bilateral internal thoracic arteries increases the number of anastomoses per bilateral thoracic artery, as well as the flexibility of the right internal thoracic artery. PMID- 11044327 TI - Marrow stromal cells for cellular cardiomyoplasty: feasibility and potential clinical advantages. AB - OBJECTIVES: Marrow stromal cells are mesenchymal stem cells able to differentiate into cardiomyocytes in vitro. We tested the hypothesis that marrow stromal cells, when implanted into myocardium, can undergo milieu-dependent differentiation and express cardiomyogenic phenotypes in vivo. METHODS: Isogenic adult rats were used as donors and recipients to simulate autologous transplantation. Marrow stromal cells isolated from donor leg bones were culture-expanded, labeled with 4;,6 diamidino-2-phenylindole, and then injected into the myocardium of the recipients. The hearts were harvested from 4 days to 12 weeks after implantation, and the implant sites were examined to identify the phenotypes of the labeled marrow stromal cells. RESULTS: Viable cells labeled with 4;, 6-diamidino-2 phenylindole can be identified in host myocardium at all time points after implantation. Implanted marrow stromal cells show the growth potential in a myocardial environment. After 4 weeks, donor cells derived from marrow stromal cells demonstrate myogenic differentiation with the expression of sarcomeric myosin heavy chain and organized contractile proteins. Positive staining for connexin 43 indicates the formation of gap junctions, which suggests that cells derived from marrow stromal cells, as well as native cardiomyocytes, are connected by intercalated disks. CONCLUSIONS: Different cell sources have been used as donor cells for cellular cardiomyoplasty. Our findings indicate that marrow stromal cells can also be used as donor cells. In an appropriate microenvironment they will exhibit cardiomyogenic phenotypes and may replace native cardiomyocytes lost by necrosis or apoptosis. Because marrow stromal cells can be obtained repeatedly by bone marrow aspiration and expanded vastly in vitro before being implanted or used as autologous implants, and because their use does not call for immunosuppression, the clinical use of marrow stromal cells for cellular cardiomyoplasty appears to be most advantageous. PMID- 11044328 TI - A novel technique for establishing total cavopulmonary connection: from surgical preconditioning to interventional completion. PMID- 11044329 TI - Actinomycosis presenting as superior vena cava syndrome in a young puerperal woman. PMID- 11044330 TI - Upper sleeve lobectomy for lung cancer with tracheal bronchus. PMID- 11044331 TI - Cardiopulmonary bypass temperature and extension of intraoperative brain damage: controversies persist. PMID- 11044332 TI - Cardiopulmonary bypass temperature and extension of intraoperative brain damage: controversies persist PMID- 11044348 TI - New treatments for advanced cancer: an approach to prioritization. AB - The allocation of funding for new anticancer treatments within the UK has not kept pace with demand. Clinicians find themselves restricted in the use of licensed drugs which they feel are in the best interests of individual patients. Against this, health authorities have a duty to ensure that scarce resources are used equitably to meet the needs of the local population as a whole. Differential levels of funding for new treatments across the country have led to concerns about rationing by postcode. This paper outlines an approach to the prioritization of new treatment for advanced cancer developed jointly by clinicians and health authorities in South London. The approach involves evidence reviews and consensus meetings. Existing and new treatments are rated on a four point 'relative effectiveness scale', which takes account of the impact of the treatment on quality of life and on survival. The strength of evidence supporting each effectiveness rating is also classified. Health Authorities have used these ratings to determine overall funding levels, while leaving decisions on individual patients to the relevant Trusts. PMID- 11044347 TI - Anxiety in cancer patients. AB - Anxiety is common in cancer patient populations, and must often initially be recognized and managed by cancer care professionals. This article reviews the recent oncology and mental health literature on anxiety. The aim is to help those involved in cancer patient care who are not specialists in mental health to understand the nature of anxiety, and discriminate morbid from normal anxiety. We review recent research into the association of anxiety with events during diagnosis and management of cancer, highlighting the importance of the meaning of events to an individual as an important factor in making people anxious. Lastly we review management strategies which might be used by cancer care professionals, in particular the importance of an awareness of specific patterns of communication which may alleviate or maintain anxiety for some cancer patients. PMID- 11044349 TI - Long-term follow-up of residual masses after chemotherapy in patients with non seminomatous germ cell tumours. AB - This retrospective study was undertaken to determine the outcome of patients with non-seminomatous germ cell tumour who achieved a serological complete response but who had residual radiologic abnormalities upon completion of primary platinum based chemotherapy. This was an analysis of 76 consecutive patients treated at Mount Vernon Hospital between 1983 and 1997. The patients were placed into two groups based upon whether they had surgical resection (surgery group, 48 patients) or observation (observation group, 28 patients) of residual radiologic masses on completion of initial chemotherapy (to enter the surgery group, complete surgical resection must have been achieved). The primary end-points were progression-free and overall survival. The percentage of patients alive with median follow-up 66 months was 90% for the surgery group and 80% for the observation group (P = 0.53, not significant). The percentage of patients continuously disease-free was 70% in the surgery group and 80% in the observation group (P = 0.31, not significant). In the small sub-group of patients with differentiated teratoma (TD) in the primary lesion who were observed, there was no excess risk of relapse or death. Patients who achieve a serological complete response after primary chemotherapy, but are left with /= + 2SD) were modest at 55%, 46%, and 52%, but in combination with CEA, increased substantially to 90%, 77% and 86%, respectively. We conclude that the serum IGF-II and IGFBP-2 profiles may provide insights into underlying biological mechanisms, and that serum IGFBP-2 may have an adjunct role in cancer surveillance in patients with colorectal cancer. PMID- 11044361 TI - High tPA-expression in primary melanoma of the limb correlates with good prognosis. AB - To investigate whether the course of primary melanoma disease correlates with expression of the various components of the proteolytic plasminogen activation (PA) system, immunohistochemical stainings for activators of plasminogen (tissue type (tPA) and urokinase type (uPA)), inhibitors of plasminogen activation (type 1 (PAI-1) and type 2 (PAI-2)) and the receptor for uPA (uPAR) were performed on 214 routinely processed melanoma lesions. All lesions were primary cutaneous melanomas, minimally 1.5 mm thick, and derived from patients with only local disease at the moment of diagnosis (clinically stage II (T(3-4)N(0)M(0)), American Joint Committee on Cancer). Median patient follow-up was 6.1 years. Single variables as immunohistochemical staining results (extent of tumour cell staining, pattern of tumour cell staining and for some components also staining of stromal cells), histopathological and clinical parameters as well as treatment variables were analysed in order to assess their prognostic importance, in terms of time to recurrence, time to distant metastasis and duration of survival. The extent of tPA tumour cell positivity, categorized as 0-5%, 6-50% and 51-100%, appeared to be of importance for these end-points. Lesions with 51-100% tPA positive tumour cells were found to have the best prognosis, whereas lesions with 6-50% tPA-positive tumour cells had the worst. Moreover, the prognostic significance of Breslow thickness, microscopic ulceration and sex was confirmed in this study. Multivariate analyses, incorporating these relevant factors, showed that the extent of tPA tumour cell positivity was an independent prognostic factor for distant metastasis-free interval (P = 0.012) and for the duration of survival (P = 0.043). PMID- 11044362 TI - Effect of c-Abl tyrosine kinase on the cellular response to paclitaxel-induced microtubule damage. AB - DNA damage has been shown to activate c-Abl tyrosine kinase. We now report that, in addition to DNA damage, microtubule damage induced by paclitaxel results in activation of c-Abl kinase. In 3T3 cells, the presence of c-Abl kinase increased paclitaxel-induced cell death. In Abl-proficient cells, paclitaxel produced a marked and prolonged G2/M arrest which peaked at 24 h and a rapid and marked induction of p21(WAF1)which also peaked at 24 h. In Abl-deficient cells, the G2/M arrest induced by paclitaxel was less prominent and shorter in duration and the effect of paclitaxel on p21(WAF1)expression was reduced and delayed. Paclitaxel had no effect on p53 expression and MAPK phosphorylation. These findings indicate that, in 3T3 cells, c-Abl kinase facilitates cell death and regulates G2/M arrest in response to paclitaxel-induced microtubule damage in a pathway that is dependent on p21(WAF1)and independent of MAPK activity. PMID- 11044363 TI - Inhibition of transcription factor nuclear factor-kappaB by a mutant inhibitor kappaBalpha attenuates resistance of human head and neck squamous cell carcinoma to TNF-alpha caspase-mediated cell death. AB - Tumour necrosis factor-alpha (TNF-alpha) is a cytokine that can induce cell death of different cancers via a cellular cascade of proteases, the caspases. However, TNF-alpha has been detected in tumour and serum of patients with head and neck squamous cell carcinoma (HNSCC), and tumour cell lines derived from this environment often exhibit resistance to TNF-alpha-induced cell death. Cell death mediated by TNF-alpha and caspases may be inhibited by cytoprotective genes regulated by transcription factor nuclear factor-kappaB (NF-kappaB). We recently showed that NF-kappaB is constitutively activated in HNSCC, and that inhibition of NF-kappaB by expression of a nondegradable mutant inhibitor of NF-kappaB, IkappaBalphaM, markedly decreased survival and growth of HNSCC cells in vivo. In the present study, we examined the TNF-alpha sensitivity and response of HNSCC with constitutively active NF-kappaB, and of HNSCC cells in which NF-kappaB is inhibited by stable expression of a dominant negative mutant inhibitor, IkappaBalphaM. Human lines UM-SCC-9, 11B and 38, previously shown to exhibit constitutive activation of NF-kappaB, were found to be highly resistant to growth inhibition by up to 10(4)U/ml of TNF-alpha in 5 day MTT assay. These TNF-alpha resistant HNSCC lines expressed TNF receptor I, and exhibited constitutive and TNF-alpha-inducible activation of NF-kappaB as demonstrated by nuclear localization of NF-kappaB p65 by immunohistochemistry. UM-SCC-9 I11 cells which stably expressed an inhibitor of NF-kappaB, IkappaBalpham, were susceptible to TNF-alpha-induced growth inhibition. DNA cell cycle analysis revealed that TNF alpha induced growth inhibition was associated with accumulation of cells with sub-G0/G1 DNA content. Cell death was demonstrated by trypan blue staining, and was blocked by caspase inhibitor. We conclude that HNSCC that exhibit constitutive and TNF-alpha-inducible activation of transcription factor NF-kappaB are resistant to TNF-alpha, and that inhibition of NF-kappaB sensitizes HNSCC to TNF-alpha caspase-mediated cytotoxicity. The demonstration of the role of activation of NF-kappaB in resistance of HNSCC to TNF-alpha may be helpful in the identification of potential targets for pharmacological, molecular and immune therapy of HNSCC. PMID- 11044364 TI - Demonstration of highly specific toxicity of the alpha-emitting radioimmunoconjugate(211)At-rituximab against non-Hodgkin's lymphoma cells. AB - The ability of an alpha-emitter conjugated to a chimaeric anti-CD20 monoclonal antibody to kill selectively human B-lymphoma cells in vitro is reported. Two B lymphoma cell lines RAEL and K422, and normal haematopoietic progenitor cells from human bone marrow aspirates were incubated with(211)At-rituximab (Rituxan(R) or MabTheratrade mark) and plated in clonogenic assays for survival analyses. Following 1 h incubation with(211)At-rituximab, in concentrations which gave an initial activity of 50 kBq ml(-1), a high tumour cell to normal bone marrow cell toxicity ratio was obtained; 4.1 to 1.0 log cell kill. Biodistribution studies of(211)At-rituximab in Balb/c mice showed similar stability as that of the iodinated analogue. The data indicate that testing of(211)At-rituximab in human patients is warranted. PMID- 11044365 TI - Bcl-2/Bax protein ratio predicts 5-fluorouracil sensitivity independently of p53 status. AB - p53 tumour-suppressor gene is involved in cell growth control, arrest and apoptosis. Nevertheless cell cycle arrest and apoptosis induction can be observed in p53-defective cells after exposure to DNA-damaging agents such as 5 fluorouracil (5-FU) suggesting the importance of alternative pathways via p53 independent mechanisms. In order to establish relationship between p53 status, cell cycle arrest, Bcl-2/Bax regulation and 5-FU sensitivity, we examined p53 mRNA and protein expression and p53 protein functionality in wild-type (wt) and mutant (mt) p53 cell lines. p53 mRNA and p53 protein expression were determined before and after exposure to equitoxic 5-FU concentration in six human carcinoma cell lines differing in p53 status and displaying marked differences in 5-FU sensitivity, with IC(50)values ranging from 0.2-22.6 mM. 5-FU induced a rise in p53 mRNA expression in mt p53 cell lines and in human papilloma virus positive wt p53 cell line, whereas significant decrease in p53 mRNA expression was found in wt p53 cell line. Whatever p53 status, 5-FU altered p53 transcriptional and translational regulation leading to up-regulation of p53 protein. In relation with p53 functionality, but independently of p53 mutational status, after exposure to 5-FU equitoxic concentration, all cell lines were able to arrest in G1. No relationship was evidenced between G1 accumulation ability and 5-FU sensitivity. Moreover, after 5-FU exposure, Bax and Bcl-2 proteins regulation was under p53 protein control and a statistically significant relationship (r = 0.880, P = 0.0097) was observed between Bcl-2/Bax ratio and 5-FU sensitivity. In conclusion, whatever p53 status, Bcl-2 or Bax induction and Bcl-2/Bax protein ratio were correlated to 5-FU sensitivity. PMID- 11044366 TI - Membrane-type 1 matrix metalloproteinase-mediated progelatinase A activation in non-tumorigenic and tumorigenic human keratinocytes. AB - Elevated expression of type IV collagenases (MMP-2 and MMP-9) has been strongly correlated with tumour progression and metastasis in various tumours. Here, we analysed expression and activation of these MMPs in non-tumourigenic HaCaT cells and the malignant HaCaT variant II-4(rt). In monolayer cultures, both cell types secreted latent MMP-2 (proMMP-2) in comparable amounts, while MMP-9 production was clearly higher in II-4(rt)cells. Upon contact with fibrillar collagen type I the malignant II-4(rt)cells, but not the HaCaT cells, gained the capability to activate proMMP-2. This process is shown to be membrane-associated and mediated by MT1-MMP. Surprisingly, all membrane preparations from either HaCaT cells or II 4(rt)cells grown as monolayers, as well as within collagen gels, contained considerable amounts of active MT1-MMP. However, within collagen gels HaCaT cells showed significantly higher TIMP-2 levels compared to II-4(rt)cells. This indicates that TIMP-2 might play a central role for MT1-MMP-mediated gelatinolytic activity. Indeed, collagen type I-induced MT1-MMP-mediated proMMP-2 activation by II-4(rt)membranes could be completely abolished by an excess of TIMP-2. In conclusion, our data suggest that MT1-MMP-mediated proMMP-2 activation might be associated with malignant progression of epidermal tumour cells. PMID- 11044367 TI - Expression of peroxisome proliferator-activated receptor (PPAR)gamma in gastric cancer and inhibitory effects of PPARgamma agonists. AB - Peroxisome proliferator-activated receptor (PPAR) gamma is expressed in human colon cancer, prostate cancer and breast cancer cells, and PPARgamma activation induces growth inhibition in these cells. PPARgamma expression in human gastric cancer cells, however, has not been fully investigated. We report the PPARgamma expression in human gastric cancer, and the effect of PPARgamma ligands on proliferation of gastric carcinoma cell lines. Immunohistochemistry was used to demonstrate the presence of PPARgamma protein in surgically resected specimens from well differentiated, moderately differentiated and poorly differentiated adenocarcinoma. We used reverse transcription-polymerase chain reaction and Northern and Western blot analyses to demonstrate PPARgamma expression in four human gastric cancer cell lines. PPARgamma agonists (troglitazone and 15-deoxy Delta(12,14)-prostaglandin J2) showed dose-dependent inhibitory effects on the proliferation of the gastric cancer cells, and their effect was augmented by the simultaneous addition of 9- cis retinoic acid, a ligand of RXRalpha. Flow cytometry demonstrated G1 cell cycle arrest and a significant increase of annexin V-positive cells after treatment with troglitazone. These results suggest that induction of apoptosis together with G1 cell cycle arrest may be one of the mechanisms of the antiproliferative effect of PPARgamma activation in human gastric cancer cells. PMID- 11044368 TI - Genomic organization and gene function in Leishmania. AB - Sequencing of the Leishmania major Friedlin genome is well underway with chromosome 1 (Chr1) and Chr3 having been completely sequenced, and Chr4 virtually complete. Sequencing of several other chromosomes is in progress and the complete genome sequence may be available as soon as 2003. A large proportion ( approximately 70%) of the newly identified genes remains unclassified, with many of these being potentially Leishmania- (or kinetoplastid-) specific. Most interestingly, the genes are organized into large (>100-300 kb) polycistronic clusters of adjacent genes on the same DNA strand. Chr1 contains two such clusters organized in a 'divergent' manner, i. e. the mRNAs for the two sets of genes are both transcribed towards the telomeres. Chr3 contains two 'convergent' clusters, with a single 'divergent' gene at one telomere, with the two large clusters separated by a tRNA gene. We have characterized several genes from the LD1 (Leishmania DNA 1) region of Chr35. BT1 (formerly ORFG) encodes a biopterin transporter and ORFF encodes a nuclear protein of unknown function. Immunization of mice with recombinant antigens from these genes results in significant reduction in parasite burden following Leishmania challenge. Recombinant ORFF antigen shows promise as a serodiagnostic. We have also developed a tetracycline regulated promoter system, which allows us to modulate gene expression in Leishmania. PMID- 11044369 TI - Life-cycle differentiation in Trypanosoma brucei: molecules and mutants. AB - Differentiation between bloodstream and tsetse midgut procyclic forms during the life cycle of the African trypanosome is an attractive model for the analysis of stage-regulated events. In particular, this transformation occurs synchronously, there are well-defined markers for stage-regulated processes and cell lines with specific defects in differentiation have been identified. This combination of tools, combined with the developing Trypanosoma brucei genome database is allowing its underlying controls to be investigated at the molecular and cytological levels. This paper examines some recent discoveries that illuminate some of the key events during trypanosome life-cycle progression. PMID- 11044371 TI - Genomics and post-genomics in parasitology: genome babble or a real opportunity? AB - The genome projects represent one of the most important developments in our knowledge of parasites. However, translation of this knowledge into an understanding of parasite biology and then on to drugs, vaccines and other healthcare developments for the diseases will need some elan and clarity of thought by scientists and funding organizations. Only then will the activity associated with post-genomics be turned from what I have termed 'genome babble' to real opportunities in understanding these parasites. PMID- 11044370 TI - The polymorphic telomeres of the African Trypanosome trypanosoma brucei. AB - African trypanosomes have plastic genomes with extensive variability at the chromosome ends. The genes encoding the expressed major surface protein of the infective bloodstream form stages of Trypanosoma brucei and are located at telomeres. These telomeric expression-site transcription units are turning out to be surprisingly polymorphic in structure and sequence. PMID- 11044372 TI - Serine proteases of the complement system. AB - The complement system in blood plasma is a major mediator of innate immune defence. The function of complement is to recognize, then opsonize or lyse, particulate materials, including bacteria, yeasts and other microrganisms, host cell debris and altered host cells. Recognition occurs by binding of complement proteins to charge or saccharide arrays. After recognition, a series of serine proteases is activated, culminating in the assembly of complex unstable proteases called C3/C5 convertases. These activate the complement protein C3, which acts as an opsonin. The complement serine proteases include the closely related C1r, C1s, MASPs 1-3 (80-90 kDa), C2 and Factor B (100 kDa), Factor D (25 kDa) and Factor I (85 kDa). Each of these has unusually restricted specificity and low enzymic activity. The C1r, C1s and MASP group occur as proenzymes. When activated, they are regulated, like many plasma serine proteases, by a serpin, C1-inhibitor. C2 and Factor B, however, have complex multiple regulation by a group of complement proteins called the Regulation of Complement Activation (or RCA) proteins, whereas Factors I and D appear to have no natural inhibitors. Advances in structure determination and protein-protein interaction properties are leading to a more detailed understanding of the complement-system proteases, and are indicating possible new routes for potential therapeutic control of complement. PMID- 11044373 TI - Toll-like receptors: lessons from knockout mice. AB - The Toll signalling pathway, which is required for establishment of dorsoventral polarity in Drosophila embryos, plays an important role in the response to microbial infections. Recently, Toll-like receptors (TLRs) have also been identified in mammals. TLR4 has been shown to function as the transmembrane component of the lipopolysaccharide receptor, while TLR2 recognizes peptidoglycans from Gram-positive bacteria, lipoproteins and yeast. Although various microbial cell-wall components are recognized by different receptors, all of these responses are abrogated in MyD88-deficient cells. These results show that different TLRs recognize different microbial cell-wall components, and that MyD88 is an essential signalling molecule shared among interleukin-1 receptor/Toll family members. PMID- 11044374 TI - The Toll/interleukin-1 receptor domain: a molecular switch for inflammation and host defence. AB - The pro-inflammatory cytokine interleukin-1 (IL-1) signals via the Type-I IL-1 receptor (IL-1RI), inducing an increase in the expression of many genes with roles in immunity and inflammation. The signalling pathways involve two adapter proteins, MyD88 and Tollip, which via two IL-1 receptor-associated kinases (IRAK and IRAK-2) activate transcription factors such as nuclear factor-kappa B and protein kinases such as p38 mitogen-activated protein kinase. A role for the low molecular-mass G-proteins Rac, Ras and Rap in these processes has also been indicated. IL-1RI is the founder of a diverse superfamily of receptors, which all share a cytosolic domain, termed the Toll/IL-1 receptor (TIR) domain. The superfamily can be divided broadly into three subgroups. The first of these is most similar to IL-1RI and includes the receptor for IL-18 and the Th2 cell regulator T1/ST2. The second subgroup is most similar to the Drosophila melanagaster protein Toll and includes Toll-like receptor 2 (TLR2), which is required for host defence against Gram-positive bacteria and fungi, and TLR4, which is required for lipopolysaccharide responsiveness, and thus is involved in host defence against Gram-negative bacteria. There are also a number of TLRs in plants and insects, all involved in host defence. The third subgroup contains non receptor proteins which possess a TIR domain and are cytosolic. MyD88 is a member, and it presumably complexes with IL-1RI via a TIR-TIR interaction. The other two members are proteins encoded by the vaccinia virus, A46R and A52R, which block TIR-dependent signalling. This receptor superfamily therefore appears to play a central role in inflammation and host defence against infection, pointing to the TIR domain as a critical molecular player in the innate immune response. PMID- 11044375 TI - Toll-like receptor family and signalling pathway. AB - Toll is a Drosophila gene essential for ontogenesis and anti-microbial resistance. Several orthologues of Toll have been identified and cloned in vertebrates, namely Toll-like receptors (TLRs). Human TLRs are a growing family of molecules involved in innate immunity. TLRs are characterized structurally by a cytoplasmic Toll/interleukin-1 receptor (TIR) domain and by extracellular leucine-rich repeats. TLRs characterized so far activate the MyD88/interleukin-1 receptor-associated kinase (IRAK) signalling pathway. Genetic, gene-transfer and dominant-negative approaches have involved TLR family members (TLR2 and TLR4) in Gram-positive and Gram-negative bacteria recognition and signalling. Accumulating evidence suggests that TLR2 is also involved in signalling-receptor complexes that recognize components of yeast and mycobacteria. However, the definitive roles of other TLRs are still lacking. A systematic approach has been used to determine whether different human leucocyte populations selectively or specifically express TLR mRNA. Based on expression pattern, TLR can be classified as ubiquitous (TLR1), restricted (TLR2, TLR4 and TLR5) and specific (TLR3). Expression and regulation of distinct but overlapping ligand-recognition patterns may underlie the existence of a large, seemingly redundant TLR family. Alternatively, the expression of a TLR in a single cell type may indicate a specific role for this molecule in a restricted setting. PMID- 11044376 TI - High environmental salinity induces memory enhancement and increases levels of brain angiotensin-like peptides in the crab Chasmagnathus granulatus. AB - Previous work on the brackish-water crab Chasmagnathus granulatus demonstrated that an endogenous peptide similar to angiotensin II plays a significant role in enhancing long-term memory that involves an association between context and an iterative danger stimulus (context-signal memory). The present results show that this memory enhancement could be produced by moving crabs from brackish water to sea water (33.0%) and keeping them there for at least 4 days. The possibility that such a facilitatory effect is due to osmotic stress is ruled out. Coincidentally, the level of angiotensin-II-like peptides in crab brain, measured by radioimmunoassay, increases with the length of exposure to sea water, reaching a significantly different level at the fourth day. The presence of angiotensin-II like immunoreactive material in neural structures of the supraoesophageal and eyestalk ganglia was confirmed by immunohistochemical analysis. The results are interpreted as supporting the hypothesis that exposure to water of high salinity is an external cue triggering a process mediated by angiotensins that leads to enhanced memory in these crabs. PMID- 11044377 TI - Morphometric partitioning of respiratory surfaces in amphioxus (Branchiostoma lanceolatum Pallas). AB - The anatomical diffusing factors (ADFs), defined as the ratio of surface area to the thickness of the diffusion barrier, of possible respiratory surfaces of adult amphioxus (Branchiostoma lanceolatum) were evaluated using stereological methods. The ADF is greatest for the lining of the atrium and for the skin covering the segmental muscles. Calculation of the diffusing capacities for O(2) revealed that the lining of the atrium makes up nearly 83 % of the entire diffusing capacity (8.86 x 10(-3) microl min(-1)mg(-1)kPa(-1) while the skin over the segmental muscles (9%), the skin over the metapleural fold (4%) and the gill bars (4%) are of minor importance. The diffusing capacity of surfaces lying over coelomic cavities makes up 76% of the whole diffusing capacity, which is consistent with the hypothesis that the coelom may function as a circulatory system for respiratory gases. Muscles have approximately 23% of the entire diffusing capacity, indicating that they may be self-sufficient for O(2) uptake. The diffusing capacity of the blood vessels in the gill bars is only 1% of the total. Thus, the 'gills' lack significant function as respiratory organs in amphioxus (lancelets). PMID- 11044378 TI - Does a rigid body limit maneuverability? AB - Whether a rigid body limits maneuverability depends on how maneuverability is defined. By the current definition, the minimum radius of the turn, a rigid bodied, spotted boxfish Ostracion meleagris approaches maximum maneuverability, i.e. it can spin around with minimum turning radii near zero. The radius of the minimum space required to turn is an alternative measure of maneuverability. By this definition, O. meleagris is not very maneuverable. The observed space required by O. meleagris to turn is slightly greater than its theoretical minimum but much greater than that of highly flexible fish. Agility, the rate of turning, is related to maneuverability. The median- and pectoral-fin-powered turns of O. meleagris are slow relative to the body- and caudal-fin-powered turns of more flexible fish. PMID- 11044379 TI - The optomotor response and spatial resolution of the visual system in male Xenos vesparum (Strepsiptera). AB - The Strepsiptera are an enigmatic group of parasitic insects whose phylogenetic relationships are hotly debated. Male Strepsiptera have very unusual compound eyes, in which each of a small number of ommatidia possesses a retina of at least 60 retinula cells. We analysed the optomotor response of Xenos vesparum males to determine whether spatial resolution in these eyes is limited by the interommatidial angle or by the higher resolution potentially provided by the extended array of retinula cells within each ommatidium. We find that the optomotor response in Strepsiptera has a typical bandpass characteristic in the temporal domain, with a temporal frequency optimum at 1-3 Hz. As a function of spatial wavelength, the optomotor response is zero at grating periods below 12 degrees and reaches its maximum strength at grating periods between 60 degrees and 70 degrees. To identify the combination of interommatidial angles and angular sensitivity functions that would generate such a spatial characteristic, we used motion detection theory to model the spatial tuning function of the strepsipteran optomotor response. We found the best correspondence between the measured response profile and theoretical prediction for an irregular array of sampling distances spaced around 9 degrees (half the estimated interommatidial angle) and an angular sensitivity function of approximately 50 degrees, which corresponds to the angular extent of the retina we estimated at the centre of curvature of the lens. Our behavioural data strongly suggest that, at least for the optomotor response, the resolution of the strepsipteran compound eye is limited by the ommatidial sampling array and not by the array of retinula cells within each ommatidium. We discuss the significance of these results in relation to the functional organisation of strepsipteran compound eyes, their evolution and the role of vision in these insects. PMID- 11044380 TI - Cloning and characterization of sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA) from crayfish axial muscle. Sarco/Endoplasmic Reticulum Ca(2+)-ATPase. AB - The discontinuous pattern of muscle growth during the moulting cycle of a freshwater crustacean (the crayfish Procambarus clarkii) was used as a model system to examine the regulation of the expression of Sarco/Endoplasmic Reticulum Ca(2+)-ATPase (SERCA). We describe the cloning, sequencing and characterization of a novel SERCA cDNA (3856 bp) obtained from crayfish axial abdominal muscle by reverse transcription/polymerase chain reaction (RT-PCR) followed by rapid amplification of cDNA ends (RACE). This complete sequence contains a 145 base pair (bp) noncoding region at the 5' end, a 3006 bp open reading frame coding for 1002 amino acid residues with a molecular mass of 110 kDa and 705 bp of untranslated region at the 3' end. This enzyme contains all the conserved domains found in 'P'-type ATPases, and the hydropathy profile suggests a transmembrane organization typical of other SERCAs. It exhibits 80% amino acid identity with Drosophila melanogaster SERCA, 79% identity with Artemia franciscana SERCA, 72% identity with rabbit fast-twitch muscle neonatal isoform SERCA1b, 71% identity with slow-twitch muscle isoform SERCA2 and 67% identity with SERCA3. Sequence alignment revealed that regions anchoring the cytoplasmic domain in the membrane were highly conserved and that most differences were in the NH(2) terminus, the central loop region and the COOH terminus. Northern analysis of total RNA from crayfish tissues probed with the 460 bp fragment initially isolated showed four bands (7.6, 7.0, 5.8 and 4.5 kilobases) displaying tissue-specific expression. SERCA was most abundant in muscle (axial abdominal, cardiac and stomach), where it is involved in Ca(2+) resequestration during relaxation, and in eggs, where it may be implicated in early embryogenesis. The level of SERCA mRNA expression in axial abdominal muscle varied during the moulting cycle as determined by slot blot analysis. SERCA expression was greatest during intermoult and decreased to approximately 50% of this level during pre- and postmoult. Patterns of gene expression for SERCA and other sarcomeric proteins during the crustacean moulting cycle may be regulated by ecdysteroids and/or mechanical stimulation. PMID- 11044381 TI - The membrane permeability transition in liver mitochondria of the great green goby Zosterisessor ophiocephalus (Pallas). AB - Liver mitochondria from the great green goby Zosterisessor ophiocephalus (Pallas) normally exhibit bioenergetic variables (membrane potential 165+/-7 mV; respiratory control ratio 6.6+/-0.4; ADP/O ratio 1.85+/-0.8; means +/- s.e.m., N=6) and activities of physiological transport systems (phosphate/proton symporter, adenine nucleotide antiporter, Ca(2+) electrophoretic uniporter) comparable with those of rat liver mitochondria. When incubated in the presence of Ca(2+) and an inducer agent such as phosphate, these mitochondria undergo a complete collapse of membrane potential accompanied by a large-amplitude swelling of the matrix, influx of sucrose from the incubation medium, release of endogenous Mg(2+) and K(+) (approximately 90% of the total) and of preaccumulated Ca(2+) and oxidation of endogenous pyridine nucleotides. All these phenomena, which are completely eliminated by cyclosporin A and inhibited with different efficacies by Mg(2+) and spermine, demonstrate that the induction of the permeability transition in this type of mitochondria has characteristics similar to those described in rat liver mitochondria. In contrast, the requirement for very high Ca(2+) concentrations (greater than 100 micromol l(-1) for the induction of the permeability transition represents a very important difference that distinguishes this phenomenon in fish and mammalian mitochondria. PMID- 11044382 TI - Open-sea migration of magnetically disturbed sea turtles. AB - Green turtles (Chelonia mydas) that shuttle between their Brazilian feeding grounds and nesting beaches at Ascension Island in the middle of the Atlantic Ocean are a paradigmatic case of long-distance oceanic migrants. It has been suggested that they calculate their position and the direction of their target areas by using the inclination and intensity of the earth's magnetic field. To test this hypothesis, we tracked, by satellite, green turtles during their postnesting migration from Ascension Island to the Brazilian coast more than 2000 km away. Seven turtles were each fitted with six powerful static magnets attached in such a way as to produce variable artificial fields around the turtle that made reliance on a geomagnetic map impossible. The reconstructed courses were very similar to those of eight turtles without magnets that were tracked over the same period and in the previous year, and no differences between magnetically disrupted and untreated turtles were found as regards navigational performance and course straightness. These findings show that magnetic cues are not essential to turtles making the return trip to the Brazilian coast. The navigational mechanisms used by these turtles remain enigmatic. PMID- 11044383 TI - Sources and mechanisms of inorganic carbon transport for coral calcification and photosynthesis. AB - The sources and mechanisms of inorganic carbon transport for scleractinian coral calcification and photosynthesis were studied using a double labelling technique with H(14)CO(3) and (45)Ca. Clones of Stylophora pistillata that had developed into microcolonies were examined. Compartmental and pharmacological analyses of the distribution of(45)Ca and H(14)CO(3) in the coelenteron, tissues and skeleton were performed in dark or light conditions or in the presence of various seawater HCO(3)(-) concentrations. For calcification, irrespective of the lighting conditions, the major source of dissolved inorganic carbon (DIC) is metabolic CO(2) (70-75% of total CaCO(3) deposition), while only 25-30% originates from the external medium (seawater carbon pool). These results are in agreement with the observation that metabolic CO(2) production in the light is at least six times greater than is required for calcification. This source is dependent on carbonic anhydrase activity because it is sensitive to ethoxyzolamide. Seawater DIC is transferred from the external medium to the coral skeleton by two different pathways: from sea water to the coelenteron, the passive paracellular pathway is largely sufficient, while a DIDS-sensitive transcellular pathway appears to mediate the flux across calicoblastic cells. Irrespective of the source, an anion exchanger performs the secretion of DIC at the site of calcification. Furthermore, a fourfold light-enhanced calcification of Stylophora pistillata microcolonies was measured. This stimulation was only effective after a lag of 10 min. These results are discussed in the context of light-enhanced calcification. Characterisation of the DIC supply for symbiotic dinoflagellate photosynthesis demonstrated the presence of a DIC pool within the tissues. The size of this pool was dependent on the lighting conditions, since it increased 39-fold after 3 h of illumination. Passive DIC equilibration through oral tissues between sea water and the coelenteric cavity is insufficient to supply this DIC pool, suggesting that there is an active transepithelial absorption of inorganic carbon sensitive to DIDS, ethoxyzolamide and iodide. These results confirm the presence of CO(2) concentrating mechanisms in coral cells. The tissue pool is not, however, used as a source for calcification since no significant lag phase in the incorporation of external seawater DIC was measured. PMID- 11044384 TI - Seasonal changes in physiology and development of cold hardiness in the hatchling painted turtle Chrysemys picta. AB - Hatchling painted turtles (Chrysemys picta) commonly hibernate in shallow, natal nests where winter temperatures may fall below -10 degrees C. Although hatchlings are moderately freeze-tolerant, they apparently rely on supercooling to survive exposure to severe cold. We investigated seasonal changes in physiology and in the development of supercooling capacity and resistance to inoculative freezing in hatchling Chrysemys picta exposed in the laboratory to temperatures that decreased from 22 to 4 degrees C over a 5.5 month period. For comparison, we also studied hatchling snapping turtles (Chelydra serpentina), a less cold-hardy species that usually overwinters under water. Although Chrysemys picta and Chelydra serpentina differed in some physiological responses, both species lost dry mass, catabolized lipid and tended to gain body water during the acclimation regimen. Recently hatched, 22 degrees C-acclimated Chrysemys picta supercooled only modestly (mean temperature of crystallization -6.3+/-0.2 degrees C; N=6) and were susceptible to inoculation by ice nuclei in a frozen substratum (mean temperature of crystallization -1.1+/-0.1 degrees C; N=6) (means +/- s.e.m.). In contrast, cold-acclimated turtles exhibited pronounced capacities for supercooling and resistance to inoculative freezing. The development of cold hardiness reflected the elimination or deactivation of potent endogenous ice nuclei and an elevation of blood osmolality that was due primarily to the retention of urea, but was not associated with accumulation of the polyols, sugars or amino acids commonly found in the cryoprotection systems of other animals. Also, Chrysemys picta (and Chelydra serpentina) lacked both antifreeze proteins and ice-nucleating proteins, which are used by some animals to promote supercooling and to initiate freezing at the high temperatures conducive to freezing survival, respectively. PMID- 11044385 TI - The pacemaker activity generating the intrinsic myogenic contraction of the dorsal vessel of Tenebrio molitor (Coleoptera). AB - Combined intracellular and extracellular recordings from various parts of the isolated dorsal vessel of Tenebrio molitor revealed some of the following electrophysiological properties of the heart and the aorta. (i) The wave of depolarization causing forward pulsation of the dorsal vessel was always transmitted from posterior to anterior, with a conduction velocity of 0.014 m s( 1) in the heart and 0.001 m s(-1) in the aorta when the heart rate was 60 beats min(-1). (ii) There was no pacemaker activity in the aorta. (iii) The duration of the compound action potential in the aortic muscle depended on the duration of the pacemaker action potential generated in the heart. (iv) Isolated parts of the heart continued to contract rhythmically for hours, indicating powerful pacemaker activity in individual cardiac segments. (v) There was a direct relationship between action potential duration and the length of the preceding diastolic interval. (vi) The rhythmic wave of depolarization was dependent on the influx of Ca(2+). (vii) The recovery of the electrical properties of myocardial cells that had been disrupted by sectioning was rapid. (viii) In hearts sectioned into two halves, the rhythmic pacemaker action potentials recorded simultaneously from the two isolated halves eventually drifted out of phase, but they had the same intrinsic frequency. In the light of these data, we discuss two alternative models for the generation of spontaneous rhythmic pumping movements of the heart and aorta. PMID- 11044386 TI - Cues by which Portia fimbriata, an araneophagic jumping spider, distinguishes jumping-spider prey from other prey. AB - Portia fimbriata from Queensland, Australia, is an araneophagic jumping spider (Salticidae) that includes in its predatory strategy a tactic (cryptic stalking) enabling it to prey effectively on a wide range of salticids from other genera. Optical cues used by P. fimbriata to identify the salticid species on which it most commonly preys, Jacksonoides queenslandicus, were investigated experimentally in the laboratory using odorless lures made from dead prey on which various combinations of features were altered. P. fimbriata adopted cryptic stalking only against intact salticid lures and modified lures on which the large anterior-median eyes were visible. Ordinary stalking was usually adopted when the lure did not have the anterior-median eyes visible. There was no evidence that cues from the legs of prey salticids influence the choice of stalking style of P. fimbriata, but cues from the legs do appear to influence strongly whether a prey is stalked at all. Cues from the cephalothorax and abdomen also influenced the stalking tendency, but to a lesser degree than cues from the legs. An algorithm to describe the perceptual processes of P. fimbriata when visually discriminating between salticid and non-salticid prey is discussed. PMID- 11044387 TI - Combined effects of cold exposure and sub-lethal intestinal parasites on host morphology and physiology. AB - Multiple, simultaneous demands elicit physiological and morphological responses that may jeopardize an animal's ability to respond to future challenges, especially when resources are limited. Laboratory mice (Mus musculus) experimentally infected with an intestinal nematode (Heligmosomoides polygyrus) and then exposed to cold showed phenotypic plasticity of morphological and physiological responses. The parasitized mice maintained a similar body mass to the unparasitized mice but had less body fat and showed changes in some organ masses, a greater resting metabolic rate (RMR) and a diminished glucose uptake capacity both at the site of infection and in regions of the small intestine not occupied by parasites. Cold-exposed mice had a greater RMR, less body fat, a greater glucose transport capacity and showed changes in organ masses compared with mice maintained at room temperature. The responses to cold exposure were not affected by parasitism for any dependent variable. The costs of having parasites during simultaneous cold exposure included decreased energy reserves and greater maintenance requirements, which may then decrease the energy available for future expenditures, such as reproduction. PMID- 11044388 TI - Evolution of air-breathing and central CO(2)/H(+) respiratory chemosensitivity: new insights from an old fish? AB - While little is known of the origin of air-breathing in vertebrates, primitive air breathers can be found among extant lobe-finned (Sarcopterygii) and ray finned (Actinopterygii) fish. The descendents of Sarcopterygii, the tetrapods, generate lung ventilation using a central pattern generator, the activity of which is modulated by central and peripheral CO(2)/H(+) chemoreception. Air breathing in Actinopterygii, in contrast, has been considered a 'reflexive' behaviour with little evidence for central CO(2)/H(+) respiratory chemoreceptors. Here, we describe experiments using an in vitro brainstem preparation of a primitive air-breathing actinopterygian, the longnose gar Lepisosteus osseus. Our data suggest (i) that gill and air-breathing motor patterns can be produced autonomously by the isolated brainstem, and (ii) that the frequency of the air breathing motor pattern is increased by hypercarbia. These results are the first evidence consistent with the presence of an air-breathing central pattern generator with central CO(2)/H(+) respiratory chemosensitivity in any primitive actinopterygian fish. We speculate that the origin of the central neuronal controller for air-breathing preceded the divergence of the sarcopterygian and actinopterygian lineages and dates back to a common air-breathing ancestor. PMID- 11044389 TI - Effect of brood size manipulation on offspring physiology: an experiment with passerine birds. AB - The environment experienced during ontogeny has a significant impact on the physiological condition of offspring. This, in turn, forecasts survival probabilities and future reproductive potential. Despite the prominent role that the concept of condition plays in evolutionary studies, the physiological and biochemical characters that define it remain relatively unexplored. In this study, we quantified the impact of brood size manipulations on the physiology and biochemistry of nestling tree swallows (Tachycineta bicolor) shortly before they fledged. Over two breeding seasons, we either increased or decreased the number of individuals in a brood by a single nestling. Every 2-4 days, we determined the resting rate of oxygen consumption [V(O(2))] of individuals in each brood. Growth was followed until 16 days of age, at which time, to look for potential trade offs in energy allocation, we measured total lipid mass, skeletal muscle and organ mass, indices of blood oxygen-carrying capacity and the activities of key metabolic enzymes in various tissues. Surprisingly, there was a minimal response of most characters to brood manipulation, suggesting that physiological and biochemical development is relatively invariant except perhaps under extreme conditions. Individuals reared in artificially enlarged broods, however, had a significantly lower body mass, body-size-adjusted [V(O(2))], gizzard mass and total lipid mass. These individuals also had decreased activity of cardiac 3 hydroxyacyl CoA dehydrogenase, suggesting a decreased capacity for oxidation of fatty acids. How these characters affect survival or the future adult phenotype remains unknown. PMID- 11044390 TI - HyAlx, an aristaless-related gene, is involved in tentacle formation in hydra. AB - Developmental gradients are known to play important roles in axial patterning in hydra. Current efforts are directed toward elucidating the molecular basis of these gradients. We report the isolation and characterization of HyAlx, an aristaless-related gene in hydra. The expression patterns of the gene in adult hydra, as well as during bud formation, head regeneration and the formation of ectopic head structures along the body column, indicate the gene plays a role in the specification of tissue for tentacle formation. The use of RNAi provides more direct evidence for this conclusion. The different patterns of HyAlx expression during head regeneration and bud formation also provide support for a recent version of a reaction-diffusion model for axial patterning in hydra. PMID- 11044391 TI - Encore is a member of a novel family of proteins and affects multiple processes in Drosophila oogenesis. AB - Mutations in the encore (enc) gene of Drosophila melanogaster cause one extra round of mitosis in the germline, resulting in the formation of egg chambers with extra nurse cells. In addition, enc mutations affect the accumulation of Gurken protein within the oocyte, leading to the production of ventralized eggs. Here we show that enc mutants also exhibit abnormalities in karyosome morphology, similar to other ventralizing mutants such as okra and spindle B. Unlike these mutants, however, the defects in Gurken accumulation and karyosome formation do not result from activation of a meiotic checkpoint. Furthermore, we demonstrate that the requirement for enc in these processes is temporally distinct from its role in germline mitosis. Cloning of the enc locus and generation of anti-Enc antibodies reveal that enc encodes a large novel protein that accumulates within the oocyte cytoplasm and colocalizes with grk mRNA. We argue that the enc mutant phenotypes reflect a role for Enc in the regulation of several RNA targets. PMID- 11044392 TI - PAN GU: a protein kinase that inhibits S phase and promotes mitosis in early Drosophila development. AB - Following completion of meiosis, DNA replication must be repressed until fertilization. In Drosophila, this replication block requires the products of the pan gu (png), plutonium (plu) and giant nuclei (gnu) genes. These genes also ensure that S phase oscillates with mitosis in the early division cycles of the embryo. We have identified the png gene and shown that it encodes a Ser/Thr protein kinase expressed only in ovaries and early embryos, and that the predicted extent of kinase activity in png mutants inversely correlates with the severity of the mutant phenotypes. The PLU and PNG proteins form a complex that has PNG-dependent kinase activity, and this activity is necessary for normal levels of mitotic cyclins. Our results reveal a novel protein kinase complex that controls S phase at the onset of development apparently by stabilizing mitotic cyclins. PMID- 11044393 TI - Sonic hedgehog regulates growth and morphogenesis of the tooth. AB - During mammalian tooth development, the oral ectoderm and mesenchyme coordinate their growth and differentiation to give rise to organs with precise shapes, sizes and functions. The initial ingrowth of the dental epithelium and its associated dental mesenchyme gives rise to the tooth bud. Next, the epithelial component folds to give the tooth its shape. Coincident with this process, adjacent epithelial and mesenchymal cells differentiate into enamel-secreting ameloblasts and dentin-secreting odontoblasts, respectively. Growth, morphogenesis and differentiation of the epithelium and mesenchyme are coordinated by secreted signaling proteins. Sonic hedgehog (Shh) encodes a signaling peptide which is present in the oral epithelium prior to invagination and in the tooth epithelium throughout its development. We have addressed the role of Shh in the developing tooth in mouse by using a conditional allele to remove Shh activity shortly after ingrowth of the dental epithelium. Reduction and then loss of Shh function results in a cap stage tooth rudiment in which the morphology is severely disrupted. The overall size of the tooth is reduced and both the lingual epithelial invagination and the dental cord are absent. However, the enamel knot, a putative organizer of crown formation, is present and expresses Fgf4, Wnt10b, Bmp2 and Lef1, as in the wild type. At birth, the size and the shape of the teeth are severely affected and the polarity and organization of the ameloblast and odontoblast layers is disrupted. However, both dentin- and enamel-specific markers are expressed and a large amount of tooth specific extracellular matrix is produced. This observation was confirmed by grafting studies in which tooth rudiments were cultured for several days under kidney capsules. Under these conditions, both enamel and dentin were deposited even though the enamel and dentin layers remained disorganized. These studies demonstrate that Shh regulates growth and determines the shape of the tooth. However, Shh signaling is not essential for differentiation of ameloblasts or odontoblasts. PMID- 11044394 TI - Zic3 is involved in the left-right specification of the Xenopus embryo. AB - Establishment of left-right (L-R) asymmetry is fundamental to vertebrate development. Several genes involved in L-R asymmetry have been described. In the Xenopus embryo, Vg1/activin signals are implicated upstream of asymmetric nodal related 1 (Xnr1) and Pitx2 expression in L-R patterning. We report here that Zic3 carries the left-sided signal from the initial activin-like signal to determinative factors such as Pitx2. Overexpression of Zic3 on the right side of the embryo altered the orientation of heart and gut looping, concomitant with disturbed laterality of expression of Xnr1 and Pitx2, both of which are normally expressed in the left lateral plate mesoderm. The results indicate that Zic3 participates in the left-sided signaling upstream of Xnr1 and Pitx2. At early gastrula, Zic3 was expressed not only in presumptive neuroectoderm but also in mesoderm. Correspondingly, overexpression of Zic3 was effective in the L-R specification at the early gastrula stage, as revealed by a hormone-inducible Zic3 construct. The Zic3 expression in the mesoderm is induced by activin (beta) or Vg1, which are also involved in the left-sided signal in L-R specification. These findings suggest that an activin-like signal is a potent upstream activator of Zic3 that establishes the L-R axis. Furthermore, overexpression of the zinc finger domain of Zic3 on the right side is sufficient to disturb the L-R axis, while overexpression of the N-terminal domain on the left side affects the laterality. These results suggest that Zic3 has at least two functionally important domains that play different roles and provide a molecular basis for human heterotaxy, which is an L-R pattern anomaly caused by a mutation in human ZIC3. PMID- 11044395 TI - Aberrant development of hippocampal circuits and altered neural activity in netrin 1-deficient mice. AB - Diffusible factors, including netrins and semaphorins, are believed to be important cues for the formation of neural circuits in the forebrain. Here we have examined the role of netrin 1 in the development of hippocampal connections. We show that netrin 1 and its receptor, Dcc, are expressed in the developing fimbria and in projection neurons, respectively, and that netrin 1 promotes the outgrowth of hippocampal axons in vitro via DCC receptors. We also show that the hippocampus of netrin 1-deficient mice shows a misorientation of fiber tracts and pathfinding errors, as detected with antibodies against the surface proteins TAG 1, L1 and DCC. DiI injections show that hippocampal commissural axons do not cross the midline in these mutants. Instead, when axons approach the midline, they turn ventrally and form a massive aberrant projection to the ipsilateral septum. In addition, both the ipsilateral entorhino-hippocampal and the CA3-to CA1 associational projections show an altered pattern of layer-specific termination in netrin 1-deficient mice. Finally, optical recordings with the Ca(2+) indicator Fura 2-AM show that spontaneous neuronal activity is reduced in the septum of netrin 1-mutant mice. We conclude that netrin 1 is required not only for the formation of crossed connections in the forebrain, but also for the appropriate layer-specific targeting of ipsilateral projections and for the control of normal levels of spontaneous neural activity. PMID- 11044396 TI - Autoregulation of Shh expression and Shh induction of cell death suggest a mechanism for modulating polarising activity during chick limb development. AB - The polarising region expresses the signalling molecule sonic hedgehog (Shh), and is an embryonic signalling centre essential for outgrowth and patterning of the vertebrate limb. Previous work has suggested that there is a buffering mechanism that regulates polarising activity. Little is known about how the number of Shh expressing cells is controlled but, paradoxically, the polarising region appears to overlap with the posterior necrotic zone, a region of programmed cell death. We have investigated how Shh expression and cell death respond when levels of polarising activity are altered, and show an autoregulatory effect of Shh on Shh expression and that Shh affects cell death in the posterior necrotic zone. When we increased Shh signalling, by grafting polarising region cells or applying Shh protein beads, this led to a reduction in the endogenous Shh domain and an increase in posterior cell death. In contrast, cells in other necrotic regions of the limb bud, including the interdigital areas, were rescued from death by Shh protein. Application of Shh protein to late limb buds also caused alterations in digit morphogenesis. When we reduced the number of Shh-expressing cells in the polarising region by surgery or drug-induced killing, this led to an expansion of the Shh domain and a decrease in the number of dead cells. Furthermore, direct prevention of cell death using a retroviral vector expressing Bcl2 led to an increase in Shh expression. Finally, we provide evidence that the fate of some of the Shh-expressing cells in the polarising region is to undergo apoptosis and contribute to the posterior necrotic zone during normal limb development. Taken together, these results show that there is a buffering system that regulates the number of Shh-expressing cells and thus polarising activity during limb development. They also suggest that cell death induced by Shh could be the cellular mechanism involved. Such an autoregulatory process based on cell death could represent a general way for regulating patterning signals in embryos. PMID- 11044397 TI - Evolutionary conservation of redundancy between a diverged pair of forkhead transcription factor homologues. AB - The Caenorhabditis elegans gene pes-1 encodes a transcription factor of the forkhead family and is expressed in specific cells of the early embryo. Despite these observations suggesting pes-1 to have an important regulatory role in embryogenesis, inactivation of pes-1 caused no apparent phenotype. This lack of phenotype is a consequence of genetic redundancy. Whereas a weak, transitory effect was observed upon disruption of just T14G12.4 (renamed fkh-2) gene function, simultaneous disruption of the activity of both fkh-2 and pes-1 resulted in a penetrant lethal phenotype. Sequence comparison suggests these two forkhead genes are not closely related and the functional association of fkh-2 and pes-1 was only explored because of the similarity of their expression patterns. Conservation of the fkh-2/pes-1 genetic redundancy between C. elegans and the related species C. briggsae was demonstrated. Interestingly the redundancy in C. briggsae is not as complete as in C. elegans and this could be explained by alterations of pes-1 specific to the C. briggsae ancestry. With overlapping function retained on an evolutionary time-scale, genetic redundancy may be extensive and expression pattern data could, as here, have a crucial role in characterization of developmental processes. PMID- 11044398 TI - FGF signalling controls the timing of Pax6 activation in the neural tube. AB - We have recently demonstrated that Pax6 activation occurs in phase with somitogenesis in the spinal cord. Here we show that the presomitic mesoderm exerts an inhibitory activity on Pax6 expression. This repressive effect is mediated by the FGF signalling pathway. The presomitic mesoderm displays a decreasing caudorostral gradient of FGF8, and grafting FGF8-soaked beads at the level of the neural tube abolishes Pax6 activation. Conversely, when FGF signalling is disrupted, Pax6 is prematurely activated in the neural plate. We propose that the progression of Pax6 activation in the neural tube is controlled by the caudal regression of the anterior limit of FGF activity. Hence, as part of its posteriorising activity, FGF8 downregulation acts as a switch from early (posterior) to a later (anterior) state of neural epithelial development. PMID- 11044399 TI - Inhibition of noggin expression in the dorsal neural tube by somitogenesis: a mechanism for coordinating the timing of neural crest emigration. AB - We have previously shown that axial-dependent delamination of specified neural crest cells is triggered by BMP4 and negatively regulated by noggin. Increasing activity of BMP4 towards the rostral part of the axis is achieved by graded expression of noggin in the dorsal neural tube, the latter being high opposite unsegmented mesoderm, and progressively downregulated facing epithelial and dissociating somites, coinciding in time and axial level with initial delamination of neural crest cells (Sela-Donenfeld, D. and Kalcheim, C. (1999) Development 126, 4749-4762). Here we report that this gradient-like expression of noggin in the neuroepithelium is controlled by the paraxial mesoderm. Deletion of epithelial somites prevented normal downregulation of noggin in the neural tube. Furthermore, partial ablation of either the dorsal half or only the dorsomedial portion of epithelial somites was sufficient to maintain high noggin expression. In contrast, deletion of the segmental plate had no effect. These data suggest that the dorsomedial region of developing somites produces an inhibitor of noggin transcription in the dorsal neural tube. Consistent with this notion, grafting dissociating somites in the place of the unsegmented mesoderm precociously downregulated the expression of noggin and triggered premature emigration of neural crest progenitors from the caudal neural tube. Thus, opposite the unsegmented mesoderm, where noggin expression is high in the neural tube, BMP4 is inactive and neural crest cells fail to delaminate. Upon somitogenesis and further dissociation, the dorsomedial portion of the somite inhibits noggin transcription. Progressive loss of noggin activity releases BMP4 from inhibition, resulting in crest cell emigration. We propose that this inhibitory crosstalk between paraxial mesoderm and neural primordium controls the timing of neural crest delamination to match the development of a suitable mesodermal substrate for subsequent crest migration. PMID- 11044400 TI - Regulation of the neural patterning activity of sonic hedgehog by secreted BMP inhibitors expressed by notochord and somites. AB - The secretion of Sonic hedgehog (Shh) from the notochord and floor plate appears to generate a ventral-to-dorsal gradient of Shh activity that directs progenitor cell identity and neuronal fate in the ventral neural tube. In principle, the establishment of this Shh activity gradient could be achieved through the graded distribution of the Shh protein itself, or could depend on additional cell surface or secreted proteins that modify the response of neural cells to Shh. Cells of the neural plate differentiate from a region of the ectoderm that has recently expressed high levels of BMPs, raising the possibility that prospective ventral neural cells are exposed to residual levels of BMP activity. We have examined whether modulation of the level of BMP signaling regulates neural cell responses to Shh, and thus might contribute to the patterning of cell types in the ventral neural tube. Using an in vitro assay of neural cell differentiation we show that BMP signaling markedly alters neural cell responses to Shh signals, eliciting a ventral-to-dorsal switch in progenitor cell identity and neuronal fate. BMP signaling is regulated by secreted inhibitory factors, including noggin and follistatin, both of which are expressed in or adjacent to the neural plate. Conversely, follistatin but not noggin produces a dorsal-to-ventral switch in progenitor cell identity and neuronal fate in response to Shh both in vitro and in vivo. These results suggest that the specification of ventral neural cell types depends on the integration of Shh and BMP signaling activities. The net level of BMP signaling within neural tissue may be regulated by follistatin and perhaps other BMP inhibitors secreted by mesodermal cell types that flank the ventral neural tube. PMID- 11044401 TI - Drosophila grain encodes a GATA transcription factor required for cell rearrangement during morphogenesis. AB - The genetic mechanisms controlling organ shape are largely unknown. We show that the Drosophila grain gene is required during development for shaping the adult legs and the larval posterior spiracles. Mutant legs are short and wide rather than long and thin, while the spiracles are flat instead of dome-shaped. We demonstrate that grain encodes the GATAc transcription factor. Analysis of loss of-function mutations at the cellular level indicates that grain affects organ shape by locally controlling cell rearrangement. Ectopic grain expression causes major morphogenetic movements, resulting in the invagination of the posterior segments into the embryo. This is the first gene that has been shown to affect epithelial morphogenesis by controlling cell rearrangements, and suggests a novel function for GATA transcription factors. PMID- 11044402 TI - Development of cranial parasympathetic ganglia requires sequential actions of GDNF and neurturin. AB - The neurotrophic factors that influence the development and function of the parasympathetic branch of the autonomic nervous system are obscure. Recently, neurturin has been found to provide trophic support to neurons of the cranial parasympathetic ganglion. Here we show that GDNF signaling via the RET/GFR(alpha)1 complex is crucial for the development of cranial parasympathetic ganglia including the submandibular, sphenopalatine and otic ganglia. GDNF is required early for proliferation and/or migration of the neuronal precursors for the sphenopalatine and otic ganglia. Neurturin exerts its effect later and is required for further development and maintenance of these neurons. This switch in ligand dependency during development is at least partly governed by the altered expression of GFR(&agr;) receptors, as evidenced by the predominant expression of GFR(&agr;)2 in these neurons after ganglion formation. PMID- 11044403 TI - The adhesion signaling molecule p190 RhoGAP is required for morphogenetic processes in neural development. AB - Rho GTPases direct actin rearrangements in response to a variety of extracellular signals. P190 RhoGAP (GTPase activating protein) is a potent Rho regulator that mediates integrin-dependent adhesion signaling in cultured cells. We have determined that p190 RhoGAP is specifically expressed at high levels throughout the developing nervous system. Mice lacking functional p190 RhoGAP exhibit several defects in neural development that are reminiscent of those described in mice lacking certain mediators of neural cell adhesion. The defects reflect aberrant tissue morphogenesis and include abnormalities in forebrain hemisphere fusion, ventricle shape, optic cup formation, neural tube closure, and layering of the cerebral cortex. In cells of the neural tube floor plate of p190 RhoGAP mutant mice, polymerized actin accumulates excessively, suggesting a role for p190 RhoGAP in the regulation of +Rho-mediated actin assembly within the neuroepithelium. Significantly, several of the observed tissue fusion defects seen in the mutant mice are also found in mice lacking MARCKS, the major substrate of protein kinase C (PKC), and we have found that p190 RhoGAP is also a PKC substrate in vivo. Upon either direct activation of PKC or in response to integrin engagement, p190 RhoGAP is rapidly translocated to regions of membrane ruffling, where it colocalizes with polymerized actin. Together, these results suggest that upon activation of neural adhesion molecules, the action of PKC and p190 RhoGAP leads to a modulation of Rho GTPase activity to direct several actin dependent morphogenetic processes required for normal neural development. PMID- 11044404 TI - Regulation of pancreas development by hedgehog signaling. AB - Pancreas organogenesis is regulated by the interaction of distinct signaling pathways that promote or restrict morphogenesis and cell differentiation. Previous work has shown that activin, a TGF(beta+) signaling molecule, permits pancreas development by repressing expression of Sonic hedgehog (Shh), a member of the hedgehog family of signaling molecules that antagonize pancreas development. Here we show that Indian hedgehog (Ihh), another hedgehog family member, and Patched 1 (Ptc1), a receptor and negative regulator of hedgehog activity, are expressed in pancreatic tissue. Targeted inactivation of Ihh in mice allows ectopic branching of ventral pancreatic tissue resulting in an annulus that encircles the duodenum, a phenotype frequently observed in humans suffering from a rare disorder known as annular pancreas. Shh(-)(/)(-) and Shh( )(/)(-) Ihh(+/)(-) mutants have a threefold increase in pancreas mass, and a fourfold increase in pancreatic endocrine cell numbers. In contrast, mutations in Ptc1 reduce pancreas gene expression and impair glucose homeostasis. Thus, islet cell, pancreatic mass and pancreatic morphogenesis are regulated by hedgehog signaling molecules expressed within and adjacent to the embryonic pancreas. Defects in hedgehog signaling may lead to congenital pancreatic malformations and glucose intolerance. PMID- 11044405 TI - Repressive and restrictive mesodermal interactions with gut endoderm: possible relation to Meckel's Diverticulum. AB - The midgut and hindgut endoderm of the mouse embryo give rise to the intestinal epithelium, yet it is not known how the intestinal program is chosen in contrast to other endoderm-derived cell types. Previous tissue explant studies with embryos at 8.5 to 11.5 days gestation (d) showed that when the gut mesoderm is removed from the prospective intestinal endoderm, the endoderm activates the expression of liver-specific genes such as serum albumin, demonstrating the endoderm's pluripotence. This reversible repression of liver genes does not affect the expression of the endodermal transcription factors HNF3 and GATA4, nor these factors' ability to engage target sites in chromatin. We have now found that at 13.5 d, the mesoderm gains a second inhibitory activity, resulting in the irreversible loss of expression of HNF3 (Foxa2) and GATA factors in the endoderm and the absence of factors binding to their target sites in chromatin. The second inhibitory activity causes the endoderm to lose the potential to activate a liver gene, and this restriction precedes the normal cytodifferentiation of the intestinal epithelium. In summary, two inhibitory interactions with mesoderm successively restrict the developmental potential of the gut endoderm, leading to intestinal differentiation. We also observed rare gut bud structures in midgestation embryos that appear to represent murine examples of Meckel's Diverticulum, a congenital abnormality in human development. The absence of restrictive mesodermal interactions could explain how Meckel's diverticula express diverse non-intestinal, endoderm-derived cell types. PMID- 11044406 TI - Hindbrain patterning: FGFs regulate Krox20 and mafB/kr expression in the otic/preotic region. AB - Krox20 and mafB/kr are regulatory genes involved in hindbrain segmentation and anteroposterior (AP) patterning. They are expressed in rhombomeres (r) r3/r5 and r5/r6 respectively, as well as in the r5/r6 neural crest. Since several members of the fibroblast growth factor (FGF) family are expressed in the otic/preotic region (r2-r6), we investigated their possible involvement in the regulation of Krox20 and mafB/kr. Application of exogenous FGFs to the neural tube of 4- to 7 somite chick embryos led to ectopic expression in the neural crest of the somitic hindbrain (r7 and r8) and to the extension of the Krox20- or mafB/kr-positive areas in the neuroepithelium. Application of an inhibitor of FGF signalling led to severe and specific downregulation of Krox20 and mafB/kr in the hindbrain neuroepithelium and neural crest. These data indicate that FGFs are involved in the control of regional induction and/or maintenance of Krox20 and mafB/kr expression, thus identifying a novel function for these factors in hindbrain development, besides their proposed more general role in early neural caudalisation. PMID- 11044407 TI - Placental vascularisation requires the AP-1 component fra1. AB - Fra1 is an immediate-early gene encoding a member of the AP-1 transcription factor family, which has diverse roles in development and oncogenesis. To determine the function of Fra1 in mouse development, the gene was inactivated by gene targeting. Foetuses lacking Fra1 were severely growth retarded and died between E10.0 and E10.5, owing to defects in extra-embryonic tissues. The placental labyrinth layer, where X-gal staining revealed expression of Fra1, was reduced in size and largely avascular, owing to a marked decrease in the number of vascular endothelial cells, as shown by the lack of Flk1 expression. In contrast, the spongiotrophoblast layer was unaffected and expressed the marker genes 4311 (Tpbp) and Flt1. Furthermore, mutant foetuses exhibited yolk-sac defects that may contribute to their growth retardation and lethality. Importantly, when the placental defect was rescued by injection of Fra1(-)(/)(-) ES cells into tetraploid wild-type blastocysts, Fra1(-)(/)(-) pups were obtained that were no longer growth retarded and survived up to 2 days after birth without apparent phenotypic defects. These data indicate that a defect in the extra embryonic compartment is causal to the observed lethality, and suggest that Fra1 plays a crucial role in establishing normal vascularisation of the placenta. PMID- 11044408 TI - Conditional deletion of the Bcl-x gene from erythroid cells results in hemolytic anemia and profound splenomegaly. AB - Bcl-x is a member of the Bcl2 family and has been suggested to be important for the survival and maturation of various cell types including the erythroid lineage. To define the consequences of Bcl-x loss in erythroid cells and other adult tissues, we have generated mice conditionally deficient in the Bcl-x gene using the Cre-loxP recombination system. The temporal and spatial excision of the floxed Bcl-x locus was achieved by expressing the Cre recombinase gene under control of the MMTV-LTR. By the age of five weeks, Bcl-x conditional mutant mice exhibited hyperproliferation of megakaryocytes and a decline in the number of circulating platelets. Three-month-old animals suffered from severe hemolytic anemia, hyperplasia of immature erythroid cells and profound enlargement of the spleen. We demonstrate that Bcl-x is only required for the survival of erythroid cells at the end of maturation, which includes enucleated reticulocytes in circulation. The extensive proliferation of immature erythroid cells in the spleen and bone marrow might be the result of a fast turnover of late red blood cell precursors and accelerated erythropoiesis in response to tissue hypoxia. The increase in cell death of late erythroid cells is independent from the proapoptotic factor Bax, as demonstrated in conditional double mutant mice for Bcl-x and Bax. Mice conditionally deficient in Bcl-x permitted us for the first time to study the effects of Bcl-x deficiency on cell proliferation, maturation and survival under physiological conditions in an adult animal. PMID- 11044410 TI - Activation tagging of the LEAFY PETIOLE gene affects leaf petiole development in Arabidopsis thaliana. AB - In a screen for leaf developmental mutants we have isolated an activator T-DNA tagged mutant that produces leaves without a petiole. In addition to that leafy petiole phenotype this lettuce (let) mutant shows aberrant inflorescence branching and silique shape. The LEAFY PETIOLE (LEP) gene is located close to the right border of the T-DNA insert linked with these dominant phenotypes and encodes a protein with a domain with similarity to the DNA binding domain of members of the AP2/EREBP family of transcription factors. Introduction of the activation-tagged LEP gene in wild-type plants conferred all the phenotypic aberrations mentioned above. The leafy petiole phenotype consists of a conversion of the proximal part of the leaf from petiole into leaf blade, which means that leaf development in let is disturbed along the proximodistal axis. Therefore, LEP is involved in either cell division activity in the marginal meristem or patterning along the proximodistal axis. PMID- 11044409 TI - Characterization of a novel subset of cardiac cells and their progenitors in the Drosophila embryo. AB - The Drosophila heart is a simple organ composed of two major cell types: cardioblasts, which form the simple contractile tube of the heart, and pericardial cells, which flank the cardioblasts. A complete understanding of Drosophila heart development requires the identification of all cell types that comprise the heart and the elucidation of the cellular and genetic mechanisms that regulate the development of these cells. Here, we report the identification of a new population of heart cells: the Odd skipped-positive pericardial cells (Odd-pericardial cells). We have used descriptive, lineage tracing and genetic assays to clarify the cellular and genetic mechanisms that control the development of Odd-pericardial cells. Odd skipped marks a population of four pericardial cells per hemisegment that are distinct from previously identified heart cells. We demonstrate that within a hemisegment, Odd-pericardial cells develop from three heart progenitors and that these heart progenitors arise in multiple anteroposterior locations within the dorsal mesoderm. Two of these progenitors divide asymmetrically such that each produces a two-cell mixed lineage clone of one Odd-pericardial cell and one cardioblast. The third progenitor divides symmetrically to produce two Odd-pericardial cells. All remaining cardioblasts in a hemisegment arise from two cardioblast progenitors each of which produces two cardioblasts. Furthermore, we demonstrate that numb and sanpodo mediate the asymmetric divisions of the two mixed-lineage heart progenitors noted above. PMID- 11044411 TI - The role of Xenopus dickkopf1 in prechordal plate specification and neural patterning. AB - Dickkopf1 (dkk1) encodes a secreted WNT inhibitor expressed in Spemann's organizer, which has been implicated in head induction in Xenopus. Here we have analyzed the role of dkk1 in endomesoderm specification and neural patterning by gain- and loss-of-function approaches. We find that dkk1, unlike other WNT inhibitors, is able to induce functional prechordal plate, which explains its ability to induce secondary heads with bilateral eyes. This may be due to differential WNT inhibition since dkk1, unlike frzb, inhibits Wnt3a signalling. Injection of inhibitory antiDkk1 antibodies reveals that dkk1 is not only sufficient but also required for prechordal plate formation but not for notochord formation. In the neural plate dkk1 is required for anteroposterior and dorsoventral patterning between mes- and telencephalon, where dkk1 promotes anterior and ventral fates. Both the requirement of anterior explants for dkk1 function and their ability to respond to dkk1 terminate at late gastrula stage. Xenopus embryos posteriorized with bFGF, BMP4 and Smads are rescued by dkk1. dkk1 does not interfere with the ability of bFGF to induce its immediate early target gene Xbra, indicating that its effect is indirect. In contrast, there is cross talk between BMP and WNT signalling, since induction of BMP target genes is sensitive to WNT inhibitors until the early gastrula stage. Embryos treated with retinoic acid (RA) are not rescued by dkk1 and RA affects the central nervous system (CNS) more posterior than dkk1, suggesting that WNTs and retinoids may act to pattern anterior and posterior CNS, respectively, during gastrulation. PMID- 11044412 TI - BMP and FGF regulate the development of EGF-responsive neural progenitor cells. AB - Temporal changes in progenitor cell responses to extrinsic signals play an important role in development, but little is known about the mechanisms that determine how these changes occur. In the rodent CNS, expression of epidermal growth factor receptors (EGFRs) increases during embryonic development, conferring mitotic responsiveness to EGF among multipotent stem cells. Here we show that cell-cell signaling controls this change. Whereas EGF-responsive stem cells develop on schedule in explant and aggregate cultures of embryonic cortex, co-culture with younger cortical cells delays their development. Exogenous BMP4 mimics the effect of younger cells, reversibly inhibiting changes in EGFR expression and responsiveness. Moreover, blocking endogenous BMP receptors in progenitors with a virus transducing dnBMPR1B accelerates changes in EGFR signaling. This involves a non-cell-autonomous mechanism, suggesting that BMP negatively regulates signal(s) that promote the development of EGF-responsive stem cells. FGF2 is a good candidate for such a signal, as we find that it antagonizes the inhibitory effects of younger cortical cells and exogenous BMP4. These findings suggest that a balance between antagonistic extrinsic signals regulates temporal changes in an intrinsic property of neural progenitor cells. PMID- 11044413 TI - Aspirin and salicylate: An old remedy with a new twist. PMID- 11044414 TI - Treatment of aortocoronary vein graft lesions with membrane-covered stents: A multicenter surveillance trial. AB - BACKGROUND: Stent implantation in lesions of degenerated aortocoronary vein grafts is associated with a high risk of periprocedural thrombus embolization and in-stent restenosis. METHODS AND RESULTS: In a multicenter study, we followed up 109 consecutive patients (mean age 66+/-8 years, 12% female) who received polytetrafluoroethylene (PTFE) membrane-covered stents for 125 de novo stenoses in vein grafts 11+/-5 years after bypass surgery. Stent deployment was successful in all but 1 patient; 1 patient suffered from subacute stent thrombosis. Six month cardiac mortality was 7% (8 patients), 3 patients (3%) underwent repeat bypass surgery, and 9 patients (8%) required target-lesion PTCA. Repeat angiography revealed vessel occlusions in 9% and in-stent restenosis in 8% of patients by the end of follow-up. CONCLUSIONS: Membrane-covered stents appear to be a safe and efficient treatment strategy associated with a low incidence of restenosis and target-vessel revascularization. Compared with previous studies, the investigated device is not associated with an increase in mortality or late vessel occlusions. PMID- 11044415 TI - Altered myocardial microvascular 3D architecture in experimental hypercholesterolemia. AB - BACKGROUND: Experimental hypercholesterolemia (HC) impairs intramyocardial microvascular function. However, whether this is associated with alterations in microvascular architecture remained unknown. Using a novel 3D micro-CT scanner, we tested the hypothesis that HC is associated with an alteration in the microvascular architecture. METHODS AND RESULTS: Pigs were euthanized after 12 weeks of either normal (n=6) or 2% HC (n=6) diet. The hearts were excised and the coronary arteries injected with a radiopaque contrast material. Myocardial samples were scanned with micro-CT, and 3D images were reconstructed with 21 microm cubic voxels. The myocardium was tomographically subdivided into subepicardium and subendocardium, and microvessels (<500 microm in diameter) were counted in situ within each region. In the subendocardium of HC pigs, the intramyocardial density of microvessels was significantly higher than in normal animals (1221.4+/-199.7 versus 758.3+/-90.8 vessels/cm(3), P:<0.05) because of an increase in the number of microvessels <200 microm in diameter (1214.4+/-199.7 versus 746. 6+/-101.5 vessels/cm(3), P:<0.05). The subepicardial vascular density was similar in both groups. CONCLUSIONS: -HC has differential effects on the spatial density of the subendocardial microvasculature that may play a role in regulation and/or spatial distribution of myocardial blood flow. This study also demonstrates the feasibility of studying myocardial microvascular architecture with micro-CT in pathophysiological states. PMID- 11044416 TI - TIMI risk score for ST-elevation myocardial infarction: A convenient, bedside, clinical score for risk assessment at presentation: An intravenous nPA for treatment of infarcting myocardium early II trial substudy. AB - BACKGROUND: Considerable variability in mortality risk exists among patients with ST-elevation myocardial infarction (STEMI). Complex multivariable models identify independent predictors and quantify their relative contribution to mortality risk but are too cumbersome to be readily applied in clinical practice. METHODS AND RESULTS: We developed and evaluated a convenient bedside clinical risk score for predicting 30-day mortality at presentation of fibrinolytic-eligible patients with STEMI. The Thrombolysis in Myocardial Infarction (TIMI) risk score for STEMI was created as the simple arithmetic sum of independent predictors of mortality weighted according to the adjusted odds ratios from logistic regression analysis in the Intravenous nPA for Treatment of Infarcting Myocardium Early II trial (n=14 114). Mean 30-day mortality was 6.7%. Ten baseline variables, accounting for 97% of the predictive capacity of the multivariate model, constituted the TIMI risk score. The risk score showed a >40-fold graded increase in mortality, with scores ranging from 0 to >8 (P:<0.0001); mortality was <1% among patients with a score of 0. The prognostic discriminatory capacity of the TIMI risk score was comparable to the full multivariable model (c statistic 0. 779 versus 0.784). The prognostic performance of the risk score was stable over multiple time points (1 to 365 days). External validation in the TIMI 9 trial showed similar prognostic capacity (c statistic 0.746). CONCLUSIONS: The TIMI risk score for STEMI captures the majority of prognostic information offered by a full logistic regression model but is more readily used at the bedside. This risk assessment tool is likely to be clinically useful in the triage and management of fibrinolytic-eligible patients with STEMI. PMID- 11044417 TI - Risk stratification in patients with inferior acute myocardial infarction treated by percutaneous coronary interventions: the role of admission troponin T. AB - BACKGROUND: Cardiac troponin T (cTnT) elevations on admission indicate a high risk subgroup of patients with ST-segment elevation acute myocardial infarction (AMI). This finding has been attributed to less effective reperfusion after thrombolytic therapy. The aim of this study was to determine the role of admission cTnT on the efficacy of percutaneous coronary interventions (PCIs) in inferior AMI. METHODS AND RESULTS: One hundred fifty-nine consecutive patients with inferior ST-segment AMI were enrolled and followed up for a mean of 448 days. Patients were stratified by cTnT on admission. A cTnT >/=0.1 microg/L was found in 58% of patients. These patients had longer time intervals from onset of symptoms to therapy (P:<0. 001) and higher 30-day (10.8% versus 1.5%, P:=0.027) and long-term (17.2% versus 4.5%, P:=0.023) cardiac mortalities. Rates of the combined end point of death, nonfatal reinfarction, and need for repeated target vessel revascularization procedures were not different in cTnT groups (log rank, 0.69; P:=0.41). PCI was attempted in 93.3% of cTnT-positive and 98.5% cTnT negative patients (P:=0.24) but was less frequently successful in patients with cTnT >/=0.1 microg/L (77.9% versus 96.9%, P:<0.001). Coronary stenting reduced 30 day and long-term cardiac mortality, particularly among cTnT-positive patients. In a multivariate analysis, cTnT indicated an approximately 5-fold-higher risk (adjusted OR, 4.6; 95% CI, 0.79 to 27.11; P:=0.089) and was a strong albeit not independent risk predictor. CONCLUSIONS: In inferior AMI, a positive admission cTnT is associated with lower success rates of direct PCI and higher rates of cardiac events over the short and long term. These patients benefit from coronary stenting. PMID- 11044418 TI - A new role for P-selectin in shear-induced platelet aggregation. AB - BACKGROUND: P-selectin, expressed on platelets on activation, mediates rolling of platelets on endothelial cells, but its role in shear-induced platelet aggregation is not known. METHODS AND RESULTS: Platelets were exposed to either a single pulse (30 seconds) or 3 pulses (10 seconds) of high shear stress (150 to 200 dynes/cm(2)) each followed by low shear stress (10 dynes/cm(2)) for 4.5 minutes or 90 seconds, respectively, at 37 degrees C to resemble more closely in vivo conditions such as those in stenotic arteries. Under these conditions, platelet aggregation was significantly increased compared with low or high shear stress alone. Monoclonal anti-P-selectin antibodies inhibited shear-induced platelet aggregation, especially when induced by the combination of high and low shear stress, by approximately 70% and had an additive effect on the inhibition by abciximab (anti-glycoprotein (GP) IIb/IIIa antibody). However, anti-P-selectin antibody inhibited shear-induced platelet aggregation only at 37 degrees C, not at 22 degrees C, whereas abciximab inhibited shear-induced platelet aggregation at both 22 degrees C and 37 degrees C. This differential effect of anti-P selectin antibody is explained by the finding that shear-induced P-selectin expression on platelets was observed mainly at 37 degrees C. CONCLUSIONS: These results indicate that pulsatile shear stress, which resembles flow conditions in stenotic arteries, induces significantly more platelet aggregation at 37 degrees C than monophasic shear stress. Under these conditions, we show a novel role for P-selectin in platelet aggregation distinct from that of GP IIb/IIIa, which may be of importance in the initiation of thrombosis associated with atherosclerotic lesions. PMID- 11044419 TI - Novel, bedside, tissue factor-dependent clotting assay permits improved assessment of combination antithrombotic and antiplatelet therapy. AB - BACKGROUND: Because optimal use of combinations of antiplatelet and antithrombotic drugs requires improved methods for assessment of therapeutic efficacy, we developed an assay designed to increase sensitivity that is based on initiation of clotting by tissue factor in minimally altered whole blood. METHODS AND RESULTS: Blood samples were obtained from healthy subjects, and the contact pathway of coagulation was inhibited with corn trypsin inhibitor (a specific factor XIIa inhibitor without effect on other coagulation factors). Clotting was initiated with relipidated tissue factor and detected with a Hemochron ACT instrument. Results were reproducible with samples from 25 healthy volunteers (mean time to clot, 125+/-17 seconds). Blood was also exposed to pharmacological concentrations of antithrombotic and antiplatelet agents in vitro. Heparin (0.25 anti-IIa/Xa U/mL) prolonged the time to clot by 2.4-fold (172 seconds, P:<0.05); hirudin (1.0 anti-IIa U/mL), by 3-fold (250 seconds P:<0.05); and enoxaparin (0.6 anti-Xa U/mL), by 2 -fold (123 seconds, P:<0.05). Additive effects of antiplatelet agents were readily detectable with both heparin and hirudin. Thus, addition of 3 microg/mL abciximab to 1.0 anti-IIa/Xa U/mL heparin and to 1.0 anti IIa U/mL hirudin further prolonged the times to clot by 140 and 67 seconds, respectively (P:<0.05 for each). Addition of abciximab to enoxaparin did not further prolong the time to clot (increment, 13 seconds; P:=NS). CONCLUSIONS: The assay developed should facilitate improved dose selection, titration, and monitoring of combination antithrombotic and antiplatelet treatment regimens. PMID- 11044420 TI - Epidemiological and genetic associations of activated factor XII concentration with factor VII activity, fibrinopeptide A concentration, and risk of coronary heart disease in men. AB - BACKGROUND: The relations of plasma activated factor XII (FXIIa) concentration and a common polymorphism (C46T) of the factor XII gene with hemostatic status and risk of coronary heart disease (CHD) were examined by prospective surveillance. METHODS AND RESULTS: Genotyping for the C46T variant was performed in 2624 men 50 to 61 years of age who were free of CHD at baseline. The genotype distribution was as follows: CC, 56.7%; CT; 36.9%; and TT, 6.6%. Plasma FXIIa was measured by ELISA on 1745 samples collected 1 year after baseline; median levels were (ng/mL) CC, 2.0; CT, 1.4; and TT, 0.8 (P:<0.0001). Respective values for plasma fibrinopeptide A (FPA, nmol/L) were 1.52, 1.35, and 1.15 (P:<0.0001); for factor VII coagulant activity (FVIIc, % standard), 114.5, 116.2, and 109.3 (P:=0.02). Group differences in FVIIc were unchanged by adjustment for body mass index and serum triglycerides. Whereas CHD incidence did not differ significantly by genotype, rates (per 1000 person-years) by thirds of FXIIa distribution were for <1.5 ng/mL, 7. 2; for 1.5 to 2.0 ng/mL, 7.2; and for >2.0 ng/mL, 13.6. Respective hazard ratios with the low third as reference group were 1.01 and 1. 96 (P:=0.007), which were essentially unchanged after allowance for genotype, blood lipids, blood pressure, body mass index, FVIIc, and FPA. CONCLUSIONS: The C46T polymorphism is a determinant of FXIIa, FPA, and possibly FVIIc, suggesting that FXII influences the activity state of the coagulation pathway and FPA cleavage from fibrinogen in vivo. Plasma FXIIa is increased in middle-aged men at high risk of CHD. PMID- 11044421 TI - Considerable time from the onset of plaque rupture and/or thrombi until the onset of acute myocardial infarction in humans: coronary angiographic findings within 1 week before the onset of infarction. AB - BACKGROUND: It has been thought that the thrombi and bleeding in plaques that occur after plaque rupture or endothelial damage from vessels with mild stenosis suddenly occlude the lumen and cause acute myocardial infarction (AMI). However, our hypothesis is that thrombi and bleeding may not suddenly occlude the lumen. METHODS AND RESULTS: The study group consisted of 20 patients who had coronary angiograms performed within 1 week (3+/-3 days) before AMI and 20 control patients who had coronary angiograms performed 6 to 18 months (282+/-49 days) before AMI. The features of infarct-related coronary segments (IRCS) at 3 days before AMI were the presence of a significant stenosis of >50% (95% in incidence and 71+/-12% diameter stenosis) and Ambrose's type II eccentric lesions (plus multiple irregularities), an indicator of plaque rupture and/or thrombi (60% [70%]), and the features at 1 year before AMI were mild stenosis of <50% (95% incidence and 30+/-18% diameter stenosis) with rare Ambrose's type II eccentric lesions (plus multiple irregularities) (10% [10%]). The same relation was observed in each of the 4 subgroups with Q-wave infarction, non-Q-wave infarction, preceding effort angina within 1 month before AMI, and no preceding effort angina. CONCLUSIONS: The appearance of marked progression and Ambrose's type II eccentric lesion on coronary angiograms 3 days before AMI suggests the presence of a considerable time from the onset of plaque rupture and/or thrombi until the onset of AMI. These features may be predictors of AMI. The concept provides new insight into the mechanism and prevention of human AMIs. PMID- 11044422 TI - The effects of beta(1)-blockade on oxidative metabolism and the metabolic cost of ventricular work in patients with left ventricular dysfunction: A double-blind, placebo-controlled, positron-emission tomography study. AB - BACKGROUND: The mechanism for the beneficial effect of beta-blocker therapy in patients with left ventricular (LV) dysfunction is unclear, but it may relate to an energy-sparing effect that results in improved cardiac efficiency. C-11 acetate kinetics, measured using positron-emission tomography (PET), are a proven noninvasive marker of oxidative metabolism and myocardial oxygen consumption (MVO(2)). This approach can be used to measure the work-metabolic index, which is a noninvasive estimate of cardiac efficiency. METHODS AND RESULTS: The aim of this study was to determine the effect of metoprolol on oxidative metabolism and the work-metabolic index in patients with LV dysfunction. Forty patients (29 with ischemic and 11 with nonischemic heart disease; LV ejection fraction <40%) were randomized to receive metoprolol or placebo in a treatment protocol of titration plus 3 months of stable therapy. Seven patients were not included in analysis because of withdrawal from the study, incomplete follow-up, or nonanalyzable PET data. The rate of oxidative metabolism (k) was measured using C-11-acetate PET, and stoke volume index (SVI) was measured using echocardiography. The work metabolic index was calculated as follows: (systolic blood pressure x SVI x heart rate)/k. No significant change in oxidative metabolism occurred with placebo (k=0.061+/-0.022 to 0.054+/-0.012 per minute). Metoprolol reduced oxidative metabolism (k=0.062+/-0. 024 to 0.045+/-0.015 per minute; P:=0.002). The work metabolic index did not change with placebo (from 5.29+/-2.46 x 10(6) to 5.14+/ 2. 06 x 10(6) mm Hg. mL/m(2)), but it increased with metoprolol (from 5. 31+/ 2.15 x 10(6) to 7.08+/-2.36 x 10(6) mm Hg. mL/m(2); P:<0.001). CONCLUSIONS: Selective beta-blocker therapy with metoprolol leads to a reduction in oxidative metabolism and an improvement in cardiac efficiency in patients with LV dysfunction. It is likely that this energy-sparing effect contributes to the clinical benefits observed with beta-blocker therapy in this patient population. PMID- 11044423 TI - Sympathetic neural burst amplitude distribution: A more specific indicator of sympathoexcitation in human heart failure. AB - BACKGROUND: Human muscle sympathetic nerve activity (MSNA) is usually measured as the number of pulse-synchronous bursts in multiunit mean voltage recordings. We recently suggested burst amplitude distribution as a more sensitive indicator of altered MSNA in congestive heart failure (CHF). Here, we test whether this distribution can discriminate between different conditions with increased MSNA burst frequency and whether it reflects single vasoconstrictor fiber firing intensity. METHODS AND RESULTS: We analyzed resting multiunit MSNA in 36 CHF patients (24 with mild to moderate CHF, 12 with severe CHF investigated before and after heart transplantation), 14 patients with pituitary deficiency, 25 matched healthy control subjects, and an additional 56 healthy men with a wider age range (21 to 71 years). Pituitary deficiency was associated with increased MSNA burst frequency (60 versus 37 bursts/min in control subjects), equivalent to that in mild to moderate CHF (61 bursts/min). However, burst amplitude distribution in hypopituitary patients (median burst amplitude, 37%) did not deviate from matched control subjects (36%), whereas amplitudes increased with disease severity in CHF (43% in mild to moderate, 52% in severe) and normalized after transplantation (36%). In the larger healthy group, MSNA burst frequency increased with age, and burst amplitude distribution remained unaffected. In 8 CHF patients, single-unit firing frequency showed a close positive relationship to multiunit burst amplitude distribution (r=0.82, P:<0.01) but none to burst frequency (r=0.39, P:=0.3). CONCLUSIONS: Muscle vasoconstrictor fiber activity is better reflected by multiunit MSNA burst amplitude distribution than by burst frequency, at least in CHF. This distribution can discriminate between conditions with increased burst frequency. PMID- 11044424 TI - Electromagnetic versus fluoroscopic mapping of the inferior isthmus for ablation of typical atrial flutter: A prospective randomized study. AB - BACKGROUND: Radiofrequency catheter ablation within the tricuspid annulus inferior caval vein isthmus can cure typical atrial flutter. The target for ablation, nonetheless, is relatively wide, and standard ablation procedures may require significant exposure to radiation. METHODS AND RESULTS: A total of 50 patients (mean age, 58+/-11 years) with typical atrial flutter were prospectively randomized to receive isthmus ablation using conventional fluoroscopy for catheter navigation (group I, n=24) or electromagnetic mapping (group II, n=26). Complete bidirectional isthmus block was verified with double potential mapping. If complete isthmus block could not be achieved after 20 radiofrequency pulses or 25 minutes of fluoroscopy, the patients were switched to the other group. Eight patients from group I (33%) but only 1 patient from group II (4%) were switched. Overall, complete isthmus block was achieved in 47 of 50 patients (94%). The overall fluoroscopy time, including the placement of the diagnostic catheters, was 22.0+/-6.3 minutes in group I and 3.9+/-1.5 minutes in group II (P:<0.0001). The fluoroscopy time needed for isthmus mapping was 17.7+/-6.5 minutes in group I and 0.2+/-0.3 minutes in group II (P:<0.0001). CONCLUSIONS: Electromagnetic mapping during the induction of linear lesions for the ablation of atrial flutter permitted a highly significant reduction in exposure to fluoroscopy while maintaining high efficacy, and it allowed the time required for fluoroscopy to be reduced to levels anticipated for diagnostic electrophysiological studies. PMID- 11044425 TI - Independent association of high blood pressure and aortic atherosclerosis: A population-based study. AB - BACKGROUND: Atherosclerosis of the thoracic aorta is associated with stroke. The association between hypertension, a major risk factor for stroke, and aortic atherosclerosis has not been determined in the general population. METHODS AND RESULTS: Transesophageal echocardiography was performed in 581 subjects, a random sample of the Olmsted County (Minnesota) population aged >/=45 years participating in the Stroke Prevention: Assessment of Risk in a Community (SPARC) study. Blood pressure was assessed by multiple office measurements and 24-hour ambulatory blood pressure monitoring. The association between blood pressure variables and aortic atherosclerosis was evaluated by multiple logistic regression, adjusting for other associated variables. Among subjects with atherosclerosis, blood pressure variables associated with complex aortic atherosclerosis (protruding plaques >/=4 mm thick, mobile debris, or ulceration) were determined. Age and smoking history were independently associated with aortic atherosclerosis of any degree (P: 95% of the LLC-PK1 derived 27-kDa selenoprotein was specifically immune precipitated by the anti-rat enzyme antibody. The hybrid enzyme had a molecular mass of 54 kDa and an s(20,w) of approximately 3.5 S indicating that every native 27-kDa selenoprotein partnered with an inert rat 27-kDa subunit in a homodimer. Enzyme assembly did not depend on the presence of the N-terminal membrane anchor of the 27-kDa subunit. Direct visualization of the deiodinase dimer showed that the holoenzyme was sorted to the basolateral plasma membrane of the renal epithelial cell. PMID- 11044449 TI - Low temperature molecular adaptation of the skeletal muscle sarco(endo)plasmic reticulum Ca2+-ATPase 1 (SERCA 1) in the wood frog (Rana sylvatica). AB - We have compared the primary sequence and enzymatic properties of the sarcoplasmic reticulum Ca(2+)-ATPases from a cold-tolerant frog Rana sylvatica with those of a closely related cold-intolerant frog, Rana clamitans. Sarcoplasmic reticulum isolated from leg muscles of both species contains a major protein ( approximately 100 kDa) that reacts with a monoclonal antibody against sarco(endo)plasmic reticulum Ca(2+)-ATPase type 1 (SERCA1). The apparent molecular mass of R. sylvatica SERCA1 is 115 kDa, whereas that of R. clamitans is 105 kDa. However, the deduced amino acid sequences obtained from cDNAs do not indicate a difference in molecular weight, thus suggesting post-translational protein modification of R. sylvatica SERCA1. Comparison of the temperature dependence of both ATP hydrolysis and Ca(2+) transport indicates that R. sylvatica SERCA1 exhibits significantly lower activation energy below 20 degrees C and an approximately 2-fold greater Ca(2+)-ATPase activity near 0 degrees C. Furthermore, R. sylvatica SERCA1 exhibits simple Michaelis-Menten kinetics with ATP and Ca(2+) as opposed to the two-site ATP kinetics and positive cooperativity with Ca(2+) observed for R. clamitans and mammalian SERCA1s. Cooperativity has been linked to protein-protein interaction in SERCA1, and this property may be altered in R. sylvatica SERCA1. Primary sequence comparison shows that R. sylvatica SERCA1 exhibits seven unique amino acid substitutions, three of which are in the ATP binding domain. We also report for the first time the presence of alternative splicing in the frog, resulting in isoforms SERCA1a and SERCA1b. Thus, it appears that the low temperature muscle contractility of R. sylvatica can be explained partially by significant functional and structural differences in SERCA1. PMID- 11044450 TI - N-terminal amino acid residues mediate protein-protein interactions between DNA bound alpha /beta -type small, acid-soluble spore proteins from Bacillus species. AB - The binding of alpha/beta-type small, acid-soluble spore proteins (SASP) to DNA of spores of Bacillus species is the major determinant of DNA resistance to a variety of damaging treatments. The primary sequence of alpha/beta-type SASP is highly conserved; however, the N-terminal third of these proteins is less well conserved than the C-terminal two-thirds. To determine the functional importance of residues in the N-terminal region of alpha/beta-type SASP, variants of SspC (a minor alpha/beta-type SASP from Bacillus subtilis) with modified N termini were generated and their structural and DNA binding properties studied in vitro and in vivo. SspC variants with deletions of up to 14 residues ( approximately 20% of SspC residues) were able to bind DNA in vitro and adopted similar conformations when bound to DNA, as determined by circular dichroism spectroscopy and protein protein cross-linking. Progressive deletion of up to 11 N-terminal residues resulted in proteins with progressively lower DNA binding affinity. However, SspC(Delta)(14) (in which 14 N-terminal residues have been deleted) showed significantly higher affinity for DNA than the larger proteins, SspC(Delta)(10) and SspC(Delta)(11). The affinity of these proteins for DNA was shown to be largely dependent upon the charge of the first few N-terminal residues. These results are interpreted in the context of a model for DNA-dependent alpha/beta type SASP protein-protein interaction involving the N-terminal regions of these proteins. PMID- 11044451 TI - Identification of residues in the Staphylococcus aureus fibrinogen-binding MSCRAMM clumping factor A (ClfA) that are important for ligand binding. AB - Clumping factor A (ClfA) is a cell surface-associated protein of Staphylococcus aureus that promotes binding of this pathogen to both soluble and immobilized fibrinogen (Fg). Previous studies have localized the Fg-binding activity of ClfA to residues 221-559 within the A region of this protein. In addition, the C terminal part of the A region (residues 484-550) has been implicated as being important for Fg binding. In this study, we further investigate the involvement of this part of ClfA in the interaction of this protein with Fg. Polyclonal antibodies generated against a recombinant protein encompassing residues 500-559 of the A region inhibited the interaction of both S. aureus and recombinant ClfA with immobilized Fg in a dose-dependent manner. Using site-directed mutagenesis, two adjacent residues, Glu(526) and Val(527), were identified as being important for the activity of ClfA. S. aureus expressing ClfA containing either the E526A or V527S substitution exhibited a reduced ability to bind to soluble Fg and to adhere to immobilized Fg. Furthermore, bacteria expressing ClfA containing both substitutions were almost completely defective in Fg binding. The E526A and V527S substitutions were also introduced into recombinant ClfA (rClfA-(221-559)) expressed in Escherichia coli. The single mutant rClfA-(221-559) proteins showed a significant reduction in affinity for both immobilized Fg and a synthetic fluorescein-labeled C-terminal gamma-chain peptide compared with the wild-type protein, whereas the double mutant rClfA-(221-559) protein was almost completely defective in binding to either species. Substitution of Glu(526) and/or Val(527) did not appear to alter the secondary structure of rClfA-(221-559) as determined by far-UV circular dichroism spectroscopy. These data suggest that the C terminus of the A region may contain at least part of the Fg-binding site of ClfA and that Glu(526) and Val(527) may be involved in ligand recognition. PMID- 11044452 TI - Cytochrome c depletion upon expression of Bcl-XS. AB - We have shown previously that Bcl-XS causes acute cell death in 3T3 cells without activating caspases (Fridman, J. S., Benedict, M. A., and Maybaum, J. (1999) Cancer Res. 59, 5999-6004). In this study, we determined that the explanation for lack of caspase activation is the cellular depletion of cytochrome c. Electron microscopy revealed gross structural changes in the mitochondria of Bcl-XS expressing cells; however, cytochrome c was not detected in cytosolic fractions from these cells. Surprisingly, it was determined that cellular cytochrome c levels decreased as Bcl-XS expression levels increased. Experiments performed to eliminate other possible explanations for the lack of caspase activation showed that these 3T3 cells have a functional cytoplasmic apoptosome, a complex of proteins that form a functional trigger capable of activating the proximal caspase in an apoptotic pathway Chinnaiyan, A. M. (1999) Neoplasia 1, 5-15, as cytosolic extracts from these cells were capable of cleaving pro-caspase-9. These cells were also able to release cytochrome c from their mitochondria after appropriate stimulation, other than Bcl-XS expression (i.e. withdrawal from serum for 24 h), and initiate a cell death that is inhibited by a dominant negative caspase-9. We conclude that lack of caspase activation is due to a Bcl-XS-induced depletion of active cytochrome c, a phenomenon that represents an alternative cell death effector pathway and/or a novel mechanism for regulating caspase activation. PMID- 11044453 TI - Tyrosine phosphorylation of the beta 4 integrin cytoplasmic domain mediates Shc signaling to extracellular signal-regulated kinase and antagonizes formation of hemidesmosomes. AB - Ligation of the alpha(6)beta(4) integrin induces tyrosine phosphorylation of the beta(4) cytoplasmic domain, followed by recruitment of the adaptor protein Shc and activation of mitogen-activated protein kinase cascades. We have used Far Western analysis and phosphopeptide competition assays to map the sites in the cytoplasmic domain of beta(4) that are required for interaction with Shc. Our results indicate that, upon phosphorylation, Tyr(1440), or secondarily Tyr(1422), interacts with the SH2 domain of Shc, whereas Tyr(1526), or secondarily Tyr(1642), interacts with its phosphotyrosine binding (PTB) domain. An inactivating mutation in the PTB domain of Shc, but not one in its SH2 domain, suppresses the activation of Shc by alpha(6)beta(4). In addition, mutation of beta(4) Tyr(1526), which binds to the PTB domain of Shc, but not of Tyr(1422) and Tyr(1440), which interact with its SH2 domain, abolishes the activation of ERK by alpha(6)beta(4). Phenylalanine substitution of the beta(4) tyrosines able to interact with the SH2 or PTB domain of Shc does not affect incorporation of alpha(6)beta(4) in the hemidesmosomes of 804G cells. Exposure to the tyrosine phosphatase inhibitor orthovanadate increases tyrosine phosphorylation of beta4 and disrupts the hemidesmosomes of 804G cells expressing recombinant wild type beta(4). This treatment, however, exerts a decreasing degree of inhibition on the hemidesmosomes of cells expressing versions of beta(4) containing phenylalanine substitutions at Tyr(1422) and Tyr(1440), at Tyr(1526) and Tyr(1642), or at all four tyrosine phosphorylation sites. These results suggest that beta(4) Tyr(1526) interacts in a phosphorylation-dependent manner with the PTB domain of Shc. This event is required for subsequent tyrosine phosphorylation of Shc and signaling to ERK but not formation of hemidesmosomes. PMID- 11044454 TI - Overexpression of a mammalian ethanolamine-specific kinase accelerates the CDP ethanolamine pathway. AB - Ethanolamine kinase (EKI) is the first committed step in phosphatidylethanolamine (PtdEtn) biosynthesis via the CDP-ethanolamine pathway. We identify a human cDNA encoding an ethanolamine-specific kinase EKI1 and the structure of the EKI1 gene located on chromosome 12. EKI1 overexpression in COS-7 cells results in a 170 fold increase in ethanolamine kinase-specific activity and accelerates the rate of [3H]ethanolamine incorporation into PtdEtn as a function of the ethanolamine concentration in the culture medium. Acceleration of the CDP-ethanolamine pathway does not result in elevated cellular PtdEtn levels, but rather the excess PtdEtn is degraded to glycerophosphoethanolamine. EKI1 has negligible choline kinase activity in vitro and does not influence phosphatidylcholine biosynthesis. Acceleration of the CDP-ethanolamine pathway also does not change the rate of PtdEtn formation via the decarboxylation of phosphatidylserine. The data demonstrate the existence of separate ethanolamine and choline kinases in mammals and show that ethanolamine kinase can be a rate-controlling step in PtdEtn biosynthesis. PMID- 11044455 TI - Regulation of nuclear transport and degradation of the Xenopus cyclin-dependent kinase inhibitor, p27Xic1. AB - The regulation of the vertebrate cell cycle is controlled by the function of cyclin-dependent kinases (CDKs), cyclins, and CDK inhibitors. The Xenopus laevis kinase inhibitor, p27(Xic1) (Xic1) is a member of the p21(Cip1)/p27(Kip1)/p57(Kip2) CDK inhibitor family and inhibits CDK2-cyclin E in vitro as well as DNA replication in Xenopus egg extracts. Xic1 is targeted for degradation in interphase extracts in a manner dependent on both the ubiquitin conjugating enzyme, Cdc34, and nuclei. Here we show that ubiquitination of Xic1 occurs exclusively in the nucleus and that nuclear localization of Xic1 is necessary for its degradation. We find that Xic1 nuclear localization is independently mediated by binding to CDK2-cyclin E and by nuclear localization sequences within the C terminus of Xic1. Our results also indicate that binding of Xic1 to CDK2-cyclin E is dispensable for Xic1 ubiquitination and degradation. Moreover, we show that amino acids 180-183 of Xic1 are critical determinants of Xic1 degradation. This region of Xic1 may define a motif of Xic1 essential for recognition by the ubiquitin conjugation machinery or for binding an alternate protein required for degradation. PMID- 11044456 TI - Binding of AP2 to sorting signals is modulated by AP2 phosphorylation. AB - The two clathrin-associated adaptor complexes AP1 and AP2 are known to participate in the formation of clathrin-coated vesicles at the trans-Golgi network and at the plasma membrane. During this process adaptors are involved in the sequestration of vesicle cargo by binding to the sorting signals within the cytoplasmic domains of the cargo proteins and in the recruitment of the clathrin coat. After budding of the clathrin-coated vesicles, the clathrin and adaptors dissociate from the vesicles. Here we show that in vitro binding of AP2 to sorting signals, which is one of the initial steps in receptor-mediated endocytosis, is modulated by adaptor phosphorylation. AP2 was phosphorylated by incubating purified AP2 in the presence of ATP and dephosphorylated by incubation with alkaline phosphatase. Affinity for tyrosine-, leucine-based and noncanonical sorting motifs was 15-33 times higher for phosphorylated than for dephosphorylated AP2. Also the binding of AP2 to membranes was regulated by adaptor phosphorylation/dephosphorylation and was about 8-fold higher for phosphorylated than for dephosphorylated AP2. Moreover, AP2 isolated from cytosol is higher phosphorylated than membrane-extracted and exhibits a 5-fold higher binding affinity than AP2 extracted from membranes. Taken together these data point to a cycle of phosphorylation/dephosphorylation as a mechanism for regulating the reversible association of AP2 with membranes and sorting signals during the process of receptor-mediated endocytosis. PMID- 11044457 TI - CREM activator and repressor isoforms in human testis: sequence variations and inaccurate splicing during impaired spermatogenesis. AB - cAMP responsive element modulator (CREM) activators are specifically expressed in haploid germ cells prior to cell elongation and are essential for spermatid development in mice. Recent studies indicate that CREM activators are also involved in the process of spermatid maturation in men. Unlike the activators, CREM repressors were suggested to be absent from adult mouse and dog testes. The present work investigates CREM transcripts in human testis with normal (n = 4) and impaired spermatogenesis (n = 2). Two activator transcripts could be identified corresponding to the tau2 isoform with and without exon gamma. Interestingly, four CREM repressors could be isolated from specimens with complete spermatogenesis. These were gamma-repressor (exons B, E, F, H, I(b)), CREM DeltaC-F, beta (exons B, G, H, I(b)), CREM DeltaC-G, beta (exons B, H, I(b)), and CREM DeltaC-G, alpha (exons B, H, I(a), I(b)). These isoforms were also present in cynomolgus monkey testes. A novel CREM splice variant (CREM DeltaC-H) was detected in a specimen with spermatid maturation defect. Beyond that, inaccurate CREM splicing, giving rise to inactive transcripts, was encountered in a specimen with impaired spermatogenesis. In conclusion, several CREM repressor transcripts are present in adult human testes, and altered transcript splicing is associated with impaired spermatogenesis. PMID- 11044458 TI - CAG repeat length of the androgen receptor gene in Japanese males with cryptorchidism. AB - We have analysed the CAG repeat length in exon 1 of the androgen receptor gene in 48 Japanese males with cryptorchidism and 100 fertile Japanese males. The CAG repeat length was 23.4 +/- 0.48 (mean +/- SE) (range 16-32, median 23) in cryptorchid patients and 23.5 +/- 0.29 (range 15-32, median 23) in normal males. There was no significant difference between the two groups. The expansion of the CAG repeats in exon 1 of the androgen receptor gene is unlikely to constitute a major cause of cryptorchidism. PMID- 11044459 TI - Vitronectin is sequestered within human spermatozoa and liberated following the acrosome reaction. AB - Vitronectin plays a role in the regulation of complement and thrombin activities and in cell surface proteolysis. Vitronectin is also an intrinsic protein of human spermatozoa. Vitronectin message has been detected in whole testis poly-A mRNA and localized by in-situ reverse transcription-polymerase chain reaction to spermatocytes. The proportion of spermatozoa that express vitronectin increases following their capacitation. In this study, spermatozoa from a man of proven fertility were probed with an anti-vitronectin monoclonal antibody (VN7) before and after their permeabilization with 0.1% Triton X-100. Of fresh spermatozoa observed by confocal microscopy, 0-8% showed vitronectin staining. However, 75% of those observed displayed vitronectin following permeabilization. Serial confocal sections through the sperm head confirmed the internal localization of vitronectin. The acrosomal status of capacitated spermatozoa that expressed vitronectin was then determined. Dual colour microscopy with rhodamine-conjugated anti-vitronectin antibody and a fluorescein-conjugated antibody directed against CD46 (a complement regulatory protein expressed on the inner acrosomal membrane) revealed that only acrosome-reacted (CD46-positive) spermatozoa displayed vitronectin. Two populations of these spermatozoa were observed. Fifty-seven of 260 (22%) were CD46-positive/vitronectin-positive and 72 of 260 (28%) were CD46 positive/vitronectin-negative. No spermatozoa were CD46-negative/vitronectin positive. These results confirm that vitronectin is released from a sequestered location within the spermatozoon following the acrosome reaction. PMID- 11044460 TI - Mouse staufen genes are expressed in germ cells during oogenesis and spermatogenesis. AB - The Drosophila melanogaster staufen gene encodes an RNA binding protein (Dm Stau) required for the localization and translational repression of mRNAs within the Drosophila oocyte. In mammals translational repression is important for normal spermatogenesis in males and storage of mRNAs in the oocytes of females. In the present study we identified two mouse cDNA expressed sequence tags (ESTs), encoding proteins with significant homology to Dm Stau and used these firstly to screen a mouse kidney cDNA library and secondly to determine whether staufen mRNAs are expressed in the ovaries and testes of mice and rats. Sequence analysis of the cDNAs revealed that they originated from two different genes. Using Northern blots of RNAs from kidneys, ovaries and testes, both cDNAs hybridized to mRNA species of approximately 3 kb in all three tissues. On sections of mouse ovaries, staufen mRNA was localized specifically to oocytes. On sections of mouse testes, staufen mRNA was expressed in spermatocytes found in seminiferous tubules at stages VI-XII of the spermatogenic cycle. In conclusion, we have shown that the mammalian homologues of Dm stau are expressed in germ cells in both male and female mice, consistent with a role for these RNA binding proteins in mammalian gametogenesis. PMID- 11044461 TI - Changes in expression of vascular endothelial growth factor and angiopoietin-1 and -2 in the macaque corpus luteum during the menstrual cycle. AB - To determine the temporal expression of vascular growth factors during the lifespan of the primate corpus luteum, experiments were designed to detect mRNA for vascular endothelial growth factor (VEGF), angiopoietin (Ang)-1 and Ang-2 and to localize protein expression for VEGF in macaque luteal tissue during the menstrual cycle. Corpora lutea (n = 3-5/stage) were collected during the early (3 5 days post-luteinizing hormone surge), mid- (6-8 days), mid-late (10-12 days), and late (14-16 days) luteal phase and at menstruation (17-18 days). Reverse transcription-polymerase chain reaction products equated to cDNA for VEGF, Ang-1 and Ang-2 in all corpora lutea. VEGF mRNA levels increased (P: < 0.05) from early to mid-luteal phase and declined in the late luteal phase and at menstruation. Immunostaining for VEGF was detected in the cytoplasm of steroidogenic luteal cells, with the most intense staining in the early luteal phase. Ang-1 and Ang-2 mRNA expression was low in the early to mid-luteal phase but increased (P: < 0.05) at late luteal phase before declining at menstruation. These data suggest transcriptional control of VEGF, Ang-1 and Ang-2, as well as post-transcriptional control of VEGF, in macaque corpus luteum. Dynamic expression of angiogenic/angiostatic factors appears critical for development, maintenance and regression of the luteal microvasculature during the menstrual cycle. PMID- 11044462 TI - Oct-4 expression in inner cell mass and trophectoderm of human blastocysts. AB - The expression of the transcription factor Oct-4 is thought to be one of the decisive factors that maintain totipotency in embryonic and germ cells. In mice, oct-4 is exclusively expressed in germ cells and totipotent cells of the embryo. In humans, Oct-4 is expressed in germ cells, embryonic stem cells and whole embryos at various stages of development. However, there is limited information about the distribution of Oct-4 expression in human embryos. In an attempt to address this issue, the inner cell mass (ICM) and trophectoderm (TE) of 17 human blastocysts were separated and Oct-4 mRNA expression individually assessed by reverse transcription-polymerase chain reaction (RT-PCR). In discarded blastocysts that developed from two pronuclear zygotes, the mean Oct-4 expression was 31 times higher in totipotent ICM cells than in differentiated TE cells. This finding suggests that, in accordance with data from the mouse, Oct-4 is highly expressed in human ICM cells as opposed to TE cells; this in turn supports the hypothesis that Oct-4 plays a similar role to maintain totipotency in these two species. PMID- 11044463 TI - Evidence for the involvement of a species-specific embryonic protease in zona escape of hamster blastocysts. AB - The source and nature of zona lytic factors during zona escape of hamster blastocyts were investigated. When cultured in hamster embryo culture medium (HECM)-2h, all 8-cell embryos (n = 135) developed to zona escaped-blastocysts with complete zona lysis. In addition, 2-cell embryos, when co-cultured with zona escaping-blastocysts (at a ratio of 1:10), exhibited zona lysis. Various other embryos at the 1-8-cell stages also showed zona lysis when cultured with zona escaping blastocysts. However, zonae from mice, rats, sheep and humans were resistant to lysis under these conditions. Pronase treatment resulted in rapid zona lysis in hamsters (7 +/- 1 s), whereas in other species zona lysis was much slower: mouse (662 +/- 27 s), rat (532 +/- 16 s), sheep (120 +/- 12 s) and human (104 +/- 8 s). When cysteine protease inhibitors (antipain, leupeptin, E-64 and p hydromercuricbenzoate) were tested, they completely inhibited zona escape, while trypsin inhibitors (TLCK and SBTI) did not. Uterine zona lysin contribution in zona escape was discounted since: (i) uterine luminal flushing and endometrial extract from day 4 (the time of zona escape in vivo) pregnant females failed to lyse zonae and (ii) endogenous oocytes and transferred 2-cell embryos (to day 3 pseudopregnant recipients) were all zona-intact, while 71% of transferred blastocysts exhibited zona escape, following their recovery after 24 h. These observations suggest that a species-specific, embryonic proteolytic factor, with a cysteine protease-like activity, is involved in the zona escape of blastocysts in hamsters. PMID- 11044464 TI - Expression of sex steroid receptors and Ki-67 in the endometria of menorrhagic women: effects of intrauterine levonorgestrel. AB - The levonorgestrel-releasing intrauterine system (LNG-IUS) has proven to be the most effective medical treatment in reducing the amount of menstrual blood loss. However, the molecular mechanisms underlying menorrhagia and/or accounting for the therapeutic effect of the LNG-IUS are still obscure. In this study, we used immunohistochemistry to compare the distribution of sex steroid receptors and the proliferation marker Ki-67 in the endometria of women with and without menorrhagia before and after 6 and 12 months of treatment with an LNG-IUS. The study sample included 67 women (aged 35-49 years) who had spontaneous ovulatory cycles. In women with menorrhagia, secretory phase endometrium exhibited more proliferative activity than in women without menorrhagia. No significant differences were found in the immunoreactivity of the oestrogen or progesterone receptors in women either with or without menorrhagia suggesting that, in addition to endocrine hormones, other factors are involved in the regulation of endometrial proliferation and menstrual blood loss. A total of 35 women were treated with LNG-IUS. After 6 months use of an LNG-IUS, the immunoreactivity of both epithelial and stromal progesterone receptors, as well as those of epithelial Ki-67 declined, and no differences were detectable between the women in the menorrhagia and control groups. Breakthrough bleeding remained a problem for nine (26%) LNG-IUS users, with no association with the pre-treatment amount of bleeding. No significant differences were found in the parameters studied between the women with and without breakthrough bleeding 6 months after insertion of an LNG-IUS. PMID- 11044466 TI - Altered subcellular distribution of cadherin-5 in endothelial cells caused by the serum of pre-eclamptic patients. AB - The main clinical features of pre-eclampsia are oedema and vascular leakage. Cadherin-5 mediates endothelial cell-cell contact in the vascular endothelium and may regulate permeability as a vascular function. Therefore, we addressed the question of whether pre-eclampsia alters cadherin-5 expression and intracellular distribution. Confluent human umbilical vein endothelial cells (HUVEC) were incubated with 20% serum from patients with pre-eclampsia (n = 18), haemolysis elevated liver enzymes-low platelet syndrome (HELLP) (n = 12), pregnancy-induced hypertension (PIH) (n = 18) or normal pregnancy (n = 10). After incubation with sera from patients with pre-eclampsia, immunostaining analyses showed cadherin-5 accumulation in vesicular and tubular structures of the Golgi apparatus. Immunoblot analyses of HUVEC after pre-eclampsia serum incubation showed an increase of the stable form of cadherin-5 while degradation products decreased. Degradation of cadherin-5 takes place at the cell membrane, so this decrease may be due to a decrease of cadherin-5 in the cell membrane. The accumulation of cadherin-5 in the vesicular and tubular structures of the Golgi apparatus indicates that targeting of cadherin-5 to the plasma membrane could be disrupted. We suggest that intracellular retention of cadherin-5 caused by serum factors in patients with pre-eclampsia may decrease the number of adhesion complexes in the cell membrane, thereby contributing to endothelial dysfunction. PMID- 11044465 TI - p57(Kip2) regulates the proper development of labyrinthine and spongiotrophoblasts. AB - The cyclin-dependent kinase (cdk) inhibitor, p57 (Kip2) is a tumour suppressor candidate and a paternally-imprinted gene. In humans, the p57(Kip2) gene is located on chromosome 11p15.5, a region implicated in both sporadic cancers and Beckwith-Wiedemann syndrome. From analysis of p57(Kip2)-deficient mice, we demonstrate the relationship between trophoblastic abnormalities and p57(Kip2). Both p57(Kip2) null ((-/-)) embryos and heterozygous embryos with a maternally derived mutated allele ((+*/-)) displayed placentomegaly, as well as dysplasia of labyrinthine and spongiotrophoblasts. The number of labyrinthine trophoblasts of homozygous embryos was twice that in wild-type embryos. When we measured kinase activities of cdk in total placenta lysates by the immuno complex kinase assay, there were no differences among the genotypes. These results show that p57(Kip2) may function in the proper development of labyrinthine and spongiotrophoblasts by pathways that are not involved with regulation of cdk activities. It is, therefore, suggested that p57(Kip2) protein might have an unknown role. PMID- 11044467 TI - Prostaglandin E(2)-dependent production of latent matrix metalloproteinase-9 in cultures of human fetal membranes. AB - Studies in our laboratory have shown that structural changes in cervical biopsied fetal membranes, prior to labour, coincide with differences in the expression of the gelatinase enzyme, latent matrix metalloproteinase-9 (MMP-9). Concurrently, in vivo, there is an increase in the expression of prostaglandins, notably prostaglandin E(2) (PGE(2)), which has been shown to regulate the expression of MMPs in other systems. The aim of this study was to test the hypothesis (using an in-vitro culture model) that endogenously produced PGE(2) has a role in the elevation of MMP-9 described in vivo. Non-infected fetal membranes sampled from women undergoing elective Caesarean section were stimulated with 10% (v/v) fetal bovine serum (FBS), a known inducer of prostaglandins. This activation resulted in a time-dependent increase in the secretion of PGE(2) into the media, as determined by enzyme-linked immunosorbent assay (day 1: 19 +/- 9 pg/ml/24 h to 358 +/- 54 pg/ml/24 h by day 4). A similar pattern of secretion of latent MMP-9 was observed in parallel with the increase in PGE(2) in the same culture media (day 1: 1.63 +/- 0.17 ng/ml/24 h to 4.2 +/- 1.4 ng/ml/24 h by day 4). When both molecules were compared, a significant (P: < 0.01) positive correlation (r = 0.623) was observed. Secretion of the tissue inhibitor of MMPs-9 (TIMP-1) was not significantly different between untreated (3.07 +/- 0.266 microg/ml/24 h) and FBS treated (3. 85 +/- 0.24 microg/ml/24 h) cultures during the first 4 days in culture. Prostaglandin synthesis inhibition studies using indomethacin (100 micromol/l) resulted in a 70-80% reduction in the activated secretion of latent MMP-9. Direct PGE(2) stimulation of cultures resulted in the bell shaped dose response curve with concentrations of 1-100 nmol/l (which are within the range secreted in culture in response to FBS), stimulating significant latent MMP-9 secretion. These results suggest a link between endogenous PGE(2) and latent MMP 9 production in human fetal membranes, raising the possibility that PGE(2) has a role in the mechanism of fetal membrane structural changes and, hence, in parturition-associated membrane rupture. PMID- 11044468 TI - Changes in the expression of nitric oxide synthase in the human uterine cervix during pregnancy and parturition. AB - Nitric oxide (NO) has been proposed as a mediator of cervical ripening. We investigated the expression, using Western blotting, and localization, using immunohistochemistry, of the nitric oxide synthase (NOS) enzymes, inducible NOS (iNOS), endothelial NOS (eNOS) and neuronal NOS (bNOS) in the human cervix during pregnancy and parturition. Cervical biopsies were obtained from non-pregnant women, women in the first trimester of pregnancy, and pregnant women at term before and after the onset of labour. Each of the NOS isoforms was localized in the cervices of both non-pregnant and pregnant subjects using immunohistochemistry. iNOS expression was significantly greater in early pregnancy compared with the non-pregnant state (P: < 0.005). iNOS expression was up-regulated further in samples obtained in the third trimester compared with the first trimester. bNOS expression was greater in samples from the first trimester of pregnancy than in non-pregnant samples (P: < 0. 005), but showed no additional increase in late pregnancy or with the onset of labour. eNOS expression was increased in samples obtained in the third trimester both before (P: = 0.002) and after the onset of labour (P: < 0.002) when compared with non-pregnant samples. The increased expression of NOS isoforms in late pregnancy supports the hypothesis that NO is involved in the process of cervical ripening. PMID- 11044469 TI - Study of DNA-methylation patterns at chromosome 15q11-q13 in children born after ICSI reveals no imprinting defects. AB - The introduction of intracytoplasmic sperm injection (ICSI) has raised concern about safety in terms of a possible increase in the incidence of major congenital malformations, chromosomal aberrations or developmental problems. The possible influence of genetic imprinting on an ICSI procedure has not yet been investigated. We therefore studied the DNA-methylation status at a defined region in chromosome 15q11-q13 in 92 children born after an ICSI procedure. Imprinting defects in this region are associated with neurogenetic disorders, e.g. Angelman syndrome (AS) and Prader-Willi syndrome (PWS). Blood samples were taken directly after birth and stored at -80 degrees C. Genomic DNA purification was performed from 3-7 ml EDTA-blood. Sodium bisulphite treatment was carried out in order to distinguish methylated from unmethylated DNA by transferring the unmethylated nucleic acid cytosine into uracil and leaving the methylated cytosine unchanged. Subsequently, a methylation-specific polymerase chain reaction (M-PCR) was performed. In all 92 children (83 from ICSI with ejaculated spermatozoa and nine from ICSI with non-ejaculated spermatozoa), a regular DNA-methylation pattern was found in the PWS/AS region. In none of the children were clinical symptoms of PWS or AS present. In conclusion, the results of this study do not indicate a higher risk of DNA-methylation defects in children born after ICSI. PMID- 11044470 TI - Comprehensive chromosomal analysis of human preimplantation embryos using whole genome amplification and single cell comparative genomic hybridization. AB - Analysis of small numbers of chromosomes using interphase fluorescent in-situ hybridization (FISH) probes has revealed that 50% of human preimplantation embryos contain abnormal cells. Detection of high levels of mosaicism with so few probes has led some researchers to extrapolate that a full analysis of all 23 pairs of chromosomes would reveal that all human embryos contain a proportion of abnormal cells. However, existing cytogenetic protocols cannot achieve such an analysis due to technical limitations. We have developed a novel technique based on whole genome amplification and comparative genomic hybridization (CGH), which for the first time allows the copy number of every chromosome to be assessed in almost every cell of a cleavage-stage embryo. We have successfully analysed 64 cells (blastomeres) derived from 12 embryos and have detected unusual forms of aneuploidy, high levels of chromosomal mosaicism, non-mosaic aneuploidy and chromosome breakage. This is the first report of a comprehensive assessment of chromosome copy number in human embryos and indicates that, despite high levels of mosaicism, some embryos do have normal chromosome numbers in every cell. Such embryos may have a superior developmental potential, and their low frequency may explain correspondingly low success rates of natural and assisted conception in humans. PMID- 11044471 TI - Hepatitis C virus: the advantages of diversity. PMID- 11044472 TI - Gene therapy for XSCID: the first success of gene therapy. PMID- 11044473 TI - Growth hormone testing and the short child. PMID- 11044474 TI - Human cytochrome oxidase deficiency. AB - The human cytochrome oxidase complex is a multisubunit assembly in the inner mitochondrial membrane responsible for the terminal event in electron transport in which molecular oxygen is reduced. Various phenotypic forms of cytochrome oxidase deficiency have been recognized, the major varieties involving degeneration of the brain stem and basal ganglia (Leigh syndrome) and lactic acidemia. Others include a fatal infantile form, a benign reversible form, and forms with cardiomyopathy. Early recognition of complementation groups within, for instance, the Leigh syndrome group has recently been followed up with a description of the gene defect for three of the nuclear-encoded forms of cytochrome c oxidase (COX) deficiency. The three genes indicted, SURF1 for Leigh syndrome, COX 10 for leukodystrophy and tubulopathy, and SCO2 for the cardiomyopathic form, all have a role in the assembly of the mature cytochrome oxidase complex. The description of these gene defects and the role these genes play are discussed in terms of what can be learned about COX assembly and about the etiology of the different phenotypic forms of the disease. PMID- 11044475 TI - Mutation analysis of the GLUT2 gene in patients with Fanconi-Bickel syndrome. AB - Fanconi-Bickel syndrome (FBS) is an autosomal recessive disorder manifesting hepatorenal glycogen accumulation, Fanconi nephropathy, and impaired utilization of glucose and galactose. Several mutations in a gene encoding a glucose transporter, GLUT2, have recently been reported in patients with FBS. We performed molecular analysis on three Japanese patients and found four novel mutations: a splice-site mutation (IVS2-2A>G), a nonsense mutation (Q287X), and two missense mutations (L389P and V423E). Heterozygotes of L389P or V423E mutation from the patients' families showed renal glucosuria. These data suggested that GLUT2 gene defects may be a cause of renal glucosuria. PMID- 11044476 TI - Celiac disease diagnosis in misdiagnosed children. AB - Antiendomysial antibodies (EMA) are today considered the most sensitive and specific serological marker of celiac disease (CD). The aim of the present study was to assess the occurrence of EMA of IgG isotype in EMA IgA negative children with clinical suspicion of malabsorption and their relationship with CD. Serum EMA IgG1 determination was performed on 30 EMA IgA negative children with clinical suspicion of CD. Total serum IgA levels were further investigated. Sixty children with gastroenterological diseases other than CD were used as control disease patients and 63 healthy children were evaluated as the control group. Eighteen out of 30 children in the study showed EMA IgG1 positivity in sera and a villous height/crypt depth ratio <3:1 as index of intestinal atrophy. It is noticeable that a selective IgA deficiency was present in only 9 of 18 EMA IgG1 positive children. In addition, clinical symptoms, EMA IgG1, and mucosal atrophy disappeared after 8-10 mo on a gluten-free diet. Neither EMA IgA nor EMA IgG1 were detected in the children in the control groups. The other 12 children in study group showed no histologic abnormalities and were EMA IgG1 negative. In this study, we reveal a group of EMA IgG1 CD children without IgA deficiency. The diagnosis was based on the presence of gluten-dependent typical serological and histologic features of CD. Our data suggest that EMA IgG1 determination could be a new tool in the diagnostic workup of CD, useful in avoiding possible misdiagnosis. PMID- 11044477 TI - Safety and efficacy of succimer in toddlers with blood lead levels of 20-44 microg/dL. Treatment of Lead-Exposed Children (TLC) Trial Group. AB - Although lead encephalopathy has virtually disappeared from the United States, thousands of children still have sufficient lead exposure to produce cognitive impairment. It is not known whether treating children with blood lead levels < 45 microg/dL (2.2 microM) is beneficial and can be done with acceptable safety. We conducted a 780-child, placebo-controlled, randomized trial of up to three courses of succimer in children with blood lead levels of 20-44 microg/dL (1.0 2.1 microM). Children were aged 12-33 mo, 77% were African-American, 7% were Hispanic, and they lived in deteriorating inner city housing. Placebo-treated children had a gradual decrease in blood lead level. Succimer-treated children had an abrupt drop in blood lead level, followed by rebound. The mean blood lead level of the succimer-treated children during the 6 mo after initiation of treatment was 4.5 microg/dL (95% confidence intervals, 3.7 to 5.3 microg/dL; 0.22 microM, 0.18 to 0.26 microM) lower than that of placebo-treated children. There were more scalp rashes in succimer-treated children (3.5% versus 1.3%) and an unanticipated excess of trauma. Succimer lowers blood lead level with few side effects. The unanticipated excess of trauma requires confirmation. PMID- 11044478 TI - The renal hemodynamic effects of ibuprofen in the newborn rabbit. AB - In early childhood, nonsteroidal anti-inflammatory drugs are mainly used to either prevent or treat premature labor of the mother and patent ductus arteriosus of the newborn infant. The most frequently used prostaglandin synthesis inhibitor is indomethacin. Fetuses exposed to indomethacin in utero have been born with renal developmental defects, and in both the unborn child and the term and premature newborn this drug may compromise renal glomerular function. The latter has in the past also been observed when i.v. indomethacin or i.v. acetylsalicylic acid (aspirin) were administered to newborn rabbits. The present experiments were designed to evaluate whether ibuprofen has less renal side effects than indomethacin, as claimed. Three groups of anesthetized, ventilated, normoxemic neonatal rabbits were infused with increasing doses of ibuprofen (0.02, 0.2, 2.0 mg/kg body weight) and the following renal parameters were measured: urine volume, urinary sodium excretion, GFR, and renal plasma flow. Renal blood flow, filtration fraction, and the renal vascular resistance were calculated according to standard formulae. Intravenous ibuprofen caused a dose-dependent, significant reduction in urine volume, GFR, and renal blood flow with a fall in filtration fraction in the animals receiving the highest dose of ibuprofen (2 mg/kg body weight). There was a very steep rise in renal vascular resistance. Urinary sodium excretion decreased. These experiments in neonatal rabbits clearly show that acute i.v. doses of ibuprofen also have significant renal hemodynamic and functional side effects, not less than seen previously with indomethacin. PMID- 11044479 TI - Genetic dissection of region associated with behavioral abnormalities in mouse models for Down syndrome. AB - Two animal models of Down syndrome (human trisomy 21) with segmental trisomy for all (Ts65Dn) or part (Ts1Cje) of human chromosome 21-homologous region of mouse chromosome 16 have cognitive and behavioral abnormalities. To compare these trisomies directly and to assess the phenotypic contribution of the region of difference between them, Ts65Dn, Ts1Cje, and a new segmental trisomic (Ms1Ts65) for the region of difference (APP: to Sod1) have been generated as littermates and tested in parallel. Although the performance of Ts1Cje mice in the Morris water maze is similar to that of Ts65Dn mice, the reverse probe tests indicate that Ts65Dn is more severely affected. By contrast, the deficits of Ms1Ts65 mice are significantly less severe than those of Ts65Dn. Therefore, whereas triplication of Sod1 to Mx1 plays the major role in causing the abnormalities of Ts65Dn in the Morris water maze, imbalance of APP: to Sod1 also contributes to the poor performance. Ts65Dn mice are hyperactive and Ts1Cje mice are hypoactive; the activity of Ms1Ts65 mice is not significantly above normal. These findings indicate that genes in the Ms1Ts65 trisomic region must interact with others in the Ts1Cje region to produce hyperactivity in Ts65Dn mice. PMID- 11044480 TI - Growth hormone stimulation testing in both short and normal statured children: use of an immunofunctional assay. AB - Accurate interpretation of the results of GH stimulation tests is of pivotal importance not only in the evaluation of the etiology of growth retardation in children but also in the selection of the best candidates for GH therapy. We performed this study to test a novel immunofunctional GH ( IFGH) assay that makes use of the concept that one GH molecule dimerizes two GH receptors and compared the results with those obtained using two GH assays, the Diagnostic Systems Laboratories ELISA and a Hybritech immunoradiometric assay in 19 children with short stature undergoing routine GH stimulation testing. We also tested 13 normally statured control children to revisit the issue of what constitutes normal GH responses to stimuli, using all three assays and arginine and either L dopa or insulin-induced hypoglycemia as secretagogues. Concentrations of IGF-I, IGF binding protein-3, and acid labile subunit were measured as well. There was a significant correlation between peak IFGH and Diagnostic Systems Laboratories ELISA GH responses to stimuli (r(2) = 0.93) as well as between the Diagnostic Systems Laboratories ELISA and Hybritech immunoradiometric assay (r(2) = 0.91). There were no significant differences between the short stature and normal group in peak or mean GH concentrations regardless of the assay used; however, the IGF I, IGF binding protein-3, and acid labile subunit concentrations were substantially lower in the short stature group. There was a wide spectrum of GH concentrations in the normal group; approximately 50% of the children had peak GH concentrations <7 ng/mL, approximately 30% <5 ng/mL, and two pubertal normal subjects peaked to only 2 ng/mL with use of both the ELISA and IFGH assays. We conclude that 1) sensitive GH assays, ELISA and immunoradiometric assay, accurately detect a GH capable of generating a biologic signal comparable to an IFGH and 2) that normal GH stimulation test results can be substantially lower than previously accepted. GH-dependent growth factors may be more sensitive indicators of GH sufficiency than GH concentrations in response to pharmacologic stimuli. PMID- 11044481 TI - Growth hormone immunoreactivity does not reflect bioactivity. AB - In childhood, the largest secretory burst of GH occurs during nighttime, and consists of a complex mixture of molecular forms of GH that are thought to have different biologic activity. Standard GH assays cannot distinguish between bioactive and biologically inactive GH isoforms. To examine this relationship, overnight GH secretion was assessed by blood sampling every 30 min in 10 short prepubertal children (7 boys and 3 girls) to evaluate both the serum concentration and the biologic activity of GH. Serum GH concentrations were measured by an immunofluorometric assay and its biologic activity by the Nb2 cell bioassay. The 12-h (2000 h to 0800 h) and 6-h (2000 h to 0200 h and 0200 h to 0800 h) GH profiles were analyzed using the Pulsar program. When GH secretory pattern was measured by immunofluorometric assay, the area under the curve above the 0 line, the mean GH concentration, and the mean height of the secretory peaks were significantly higher during the first than during the second part of the night (29.17+/-5.93 versus 16.29+/-1.87 mIU/L, p<0.05; 7.77+/-1.28 versus 4.83+/ 0.33 mIU/L, p<0.05; 4.61+/-0.94 versus 2.68+/-0.27 mIU/L, p<0.05, respectively). In contrast, GH biologic activity was not significantly different during the two parts of the night. In conclusion, a dissociation between GH bioactivity and immunoreactivity is present in physiologic conditions, indicating that standard GH measurements do not provide any information on the biologic activity of the hormone. Although GH secretion is regulated by complex neuroendocrine mechanisms, the biologic activity of the hormone seems to be independent of them and is most probably regulated by peripheral mechanisms acting on its clearance or bioavailability to the target tissues. PMID- 11044482 TI - Serum lipid concentrations and growth characteristics in 12-year-old children born small for gestational age. AB - According to Barker's hypothesis, children born small for gestational age (SGA) are at increased risk for cardiovascular diseases in adulthood. The aim of our study was to determine whether retarded fetal growth is associated with dyslipidemia in childhood and, if so, to find predictive factors in the growth characteristics of SGA children. We studied the serum lipid concentrations of 55 SGA children and their 55 appropriate for gestational age control subjects at the age of 12 y. Growth variables were recorded at birth, 5 y, and 12 y of age. The study group consisted of all full-term SGA children born at our university hospital during a 22-mo period in 1984-1986. Nearly half of the SGA children (47.3%) were in the highest quartile for serum total cholesterol of the appropriate for gestational age children (p = 0.038). In multiple logistic regression analysis, poor catch-up growth in height (odds ratio, 13. 8; 95% confidence interval, 2.0-97.5), female sex (odds ratio, 8.1; 95% confidence interval, 1.3-48.9), and early stage of puberty (odds ratio, 7.5; 95% confidence interval, 1.2-46.5) predicted high cholesterol level in the SGA children. By the age of 5 y, 20 (36.4%) SGA children showed catch-up growth of > or =2 SD scores in height, and 21 (38.2%) SGA children showed catch-up growth of > or =2 SD scores in weight from birth. At the age of 12 y, the SGA children were still significantly shorter (p<0.001) and lighter (p< 0.05) than the appropriate for gestational age children, even though their pubertal development was similarly advanced. In conclusion, to be born SGA has long-term consequences for later growth and may already influence the level of serum total cholesterol before the teens. SGA children with poor catch-up growth in height may be at the highest risk for hypercholesterolemia. PMID- 11044483 TI - Adrenal function in sick very preterm infants. AB - Some very preterm neonates admitted to the neonatal intensive care unit show circulatory and respiratory problems that improve after administration of steroids. It is unclear whether these symptoms could be caused by adrenal insufficiency. The objective of our study was to investigate the cortisol levels and the cortisol release from the adrenals after ACTH in very preterm infants with and without severe illness and to find whether a relation exists between adrenal function and outcome. An ACTH test (0.5 microg) was performed on d 4 in 21 very preterm infants (gestational age, 25.6-29.6 wk; birth weight, 485-1265 g). Baseline cortisol and 17-hydroxyprogesterone (17OHP) levels and the cortisol levels 30, 60, and 120 min after ACTH administration were measured. The Score for Neonatal Acute Physiology was used to measure illness severity. All infants showed an increase in cortisol levels after ACTH, but the cortisol levels were significantly lower in the ventilated more severely ill infants. After adjusting for birth weight and gestational age, the mean baseline cortisol levels and cortisol/17OHP ratios were significantly lower and the 17OHP levels significantly higher in the ventilated infants compared with the nonventilated infants. Patients with an adverse outcome had significantly lower baseline cortisol/17OHP ratios and 60-min cortisol levels during ACTH testing (p = 0.002 and p = 0.03, respectively). These data suggest an insufficient adrenal response to stress in sick ventilated very preterm infants with gestational ages younger than 30 wk compared with nonventilated less sick preterm infants. Further studies are required to investigate whether supplementation with physiologic doses of hydrocortisone may benefit the outcome. PMID- 11044484 TI - Immunocytochemical characterization of murine Hex, a homeobox-containing protein. AB - A polyclonal antibody against a glutathione S:-transferase fusion protein containing the 76 COOH-terminal amino acids of Hex, a divergent homeobox gene, was raised in rabbits. Western blot and immunofluorescence reveal that Hex is a 35-37-kD soluble protein present both in the nucleus and cytoplasm of transfected and nontransfected cultured cells as well as in whole mouse embryo. Confocal microscopy of whole mount immunostained mouse embryos at E7. 5 and E8.5 demonstrates that Hex is differentially localized in the cytoplasm and nucleus of definitive endoderm, developing blood islands, and hepatic diverticulum. In particular, in the region of the foregut that gives rise to the liver, Hex expression is nuclear in the endodermal cells of the hepatic diverticulum, whereas expression is primarily cytoplasmic in cells lateral to the liver-forming region. This suggests that nuclear localization of Hex is involved in early hepatic specification and that compartmentalization of Hex protein plays an important role in its function during mouse development. PMID- 11044485 TI - Maximum length sequence brainstem auditory evoked responses in term neonates who have perinatal hypoxia-ischemia. AB - Maximum length sequence brainstem auditory evoked response (BAER) was studied within the first week after birth in 28 term neonates who had perinatal hypoxia ischemia, or asphyxia. In the BAER recorded using conventional averaging techniques (click rate 21/s), the only abnormality was a slight increase in III-V interval, in addition to an increase in wave latencies when including those who had an elevated threshold (t test, all p<0.05). In the maximum length sequence BAER, however, both the III-V and I-V intervals in the asphyxiated infants were significantly increased at all the 91/s, 227/s, 455/s, and particularly 910/s click rates (p<0.05-0.001). The I-III interval was also increased significantly at 455/s and 910/s click rates (both p< 0.05). Wave V amplitude was significantly reduced at all the click rates used (ANOVA, p<0.05-0.001), particularly at 910/s, which sometimes was the only abnormality indicative of brain damage. Both the amplitude ratios V/I and V/III were significantly decreased at 455/s and 910/s click rates (p<0.01 or 0.001). A general trend was that BAER abnormalities after hypoxia-ischemia became more prominent as click rate was increased. Significant abnormalities occurred mainly at very high click rates (455/s and 910/s), which can be achieved using the maximum length sequence technique but not by using conventional averaging techniques. Thus, this technique, which can be used at the cribside, appears to be a better method for the early detection of brain damage after hypoxia-ischemia than using conventional averaging techniques, enhancing the diagnostic value of the BAER. PMID- 11044486 TI - Fetal plasma leptin concentrations: relationship with different intrauterine growth patterns from 19 weeks to term. AB - The relationship between in utero fetal growth and fetal leptin concentrations was investigated between 19 and 41 wk in 40 normal (appropriate for gestational age, AGA) fetuses, in 25 intrauterine growth-restricted (IUGR) fetuses, and in 18 fetuses from gestational diabetic mothers (GDM), representing different intrauterine growth patterns. Umbilical venous plasma leptin concentrations were determined at the time of either in utero fetal blood sampling or delivery. Plasma leptin was measurable as early as 19 wk of gestation. A significant difference was observed between umbilical venous and arterial plasma leptin concentrations (0.6+/-0.6 ng/mL; p<0.01). In AGA and in IUGR fetuses, significant positive relationships were found between fetal leptin concentrations and both gestational age (p<0.001) and fetal weight (p<0.001). Leptin concentrations were significantly higher in AGA than IUGR only after 34 wk (p<0.05), but leptin per kilogram fetal weight (leptin/kg) was not significantly different. In IUGR with abnormal umbilical arterial Doppler velocimetry and fetal heart rate, leptin/kg significantly higher than in IUGR with normal biophysical and biochemical parameters was found (p<0.05). Both circulating plasma leptin and leptin/kg were significantly higher in GDM than in normal fetuses (p<0.001) and correlated with abdominal fat mass measured by ultrasound. No gender differences were observed in any group of fetuses. These findings indicate a clear relationship between fetal leptin concentrations and fetal fat mass. Data in severe IUGR suggest the presence of increased leptin concentrations associated with in utero signs of fetal distress. PMID- 11044487 TI - Activity and expression of the Na(+)/H(+) exchanger in the microvillous plasma membrane of the syncytiotrophoblast in relation to gestation and small for gestational age birth. AB - The effect of gestational age, low birth weight, and umbilical plasma pH on the activity and expression of the Na(+)/H(+) exchanger in the microvillous plasma membrane (MVM) of the placental syncytiotrophoblast was investigated. MVM were isolated from placentas of fetuses delivered in the first and second trimesters and from appropriately grown for gestational age (AGA) and small for gestational age (SGA) babies born at term. Na(+)/H(+) exchange activity (amiloride-sensitive Na(+) uptake) was higher (p<0.05) in second trimester and term AGA MVM versus first trimester MVM (median [range]: 1.80 [1.01-3.03], 1.72 [1.16-3.15] versus 1.48 [0.92-1.66] nmol/mg protein/30s, respectively, n = 6, 12, and 9). As regards exchanger isoforms, Western blotting showed that NHE1 expression did not change across gestation, but NHE2 and NHE3 expression were lower (p<0.01) in the first and second trimesters than in term AGA MVM. There were no differences in Na(+)/H(+) exchanger activity or in NHE1-3 expression in term AGA MVM versus SGA (n = 11) MVM. There was no correlation between exchanger activity and umbilical artery or vein plasma pH, although with a relatively small number of samples (n = 12 and 15, respectively). We conclude that there is differential regulation of the activity and expression of Na(+)/H(+) exchanger isoforms in the MVM over the course of gestation in normal pregnancy; this is not affected in pregnancies resulting in SGA babies at term. PMID- 11044488 TI - Origin and fate of erythropoietin in human milk. AB - Erythropoietin (Epo) is a normal constituent of human milk, but the origin and fate of this cytokine in milk are not known. Regarding its origin, we hypothesized that cells of the mammary gland secrete Epo into milk actively and, therefore, that concentrations in milk do not correlate with concentrations in serum. Regarding its fate, we hypothesized that Epo concentrations in milk change with time postpartum and that Epo in milk is protected from digestion in the neonatal gastrointestinal tract. To address these issues, we measured Epo concentrations in 103 milk samples (ELISA), 55 of which were paired with serum. Mammary duct epithelial cells were evaluated as a source of Epo by breast tissue immunohistochemistry and by cell culture. Circulating and milk Epo were compared by Western analysis to detect size differences, possibly reflecting differences in processing. Epo stability in simulated conditions of digestion was evaluated. We observed that milk Epo concentrations increase as a function of duration of breast-feeding and have a negative correlation with serum Epo or milk protein concentration. Mammary duct epithelial cells from breast biopsies of lactating women had marked immunoreactivity to Epo, but such activity was minimal to absent in nonlactating breast tissue. Further evidence that mammary duct epithelia produce Epo was obtained by observing Epo mRNA and protein expression in cultured human mammary epithelial cells. The molecular size of Epo in milk and serum is identical. Recombinant Epo added to human milk or commercial infant formulas was relatively stable in conditions that simulate gastric and small intestinal conditions of newborn infants; however, recombinant Epo added to D(5)W was not protected from digestion. We conclude that Epo concentrations in milk increase as a function of the duration of breast feeding, that Epo is actively secreted into human milk by mammary duct epithelia, and that the Epo within milk is largely protected from digestion. PMID- 11044489 TI - Perinatal feedings adversely affect lipogenic activities but not glucose handling in adult rats. AB - The long-term effects of early under- and overfeeding on glucose metabolism and fat cell lipogenesis were studied. Newborn rats were reared in litter sizes of four, 10, and 16 pups. The amount of milk intake per pup varied inversely with litter sizes. A subgroup of pups from each group was studied at age 20 d, whereas another subgroup was weaned to an ad libitum feeding of standard rat chow and studied at 12 wk of age. There were no differences among groups in food intake on the basis of per gram body weight. Overfeeding during suckling resulted in fatter rats at weaning and in the adults. The higher fat contents in the adipose tissues and carcasses were associated with higher fatty acid synthase and lipogenic activities in the adipose tissues at weaning and 12 wk of age. Differences in plasma insulin and glucose levels among groups were observed only in the 20-d-old rats: basal insulin and glucose levels and 30-min postglucose insulin levels were highest in the overnourished and lowest in the undernourished rats. However, by 12 wk of age, there were no significant differences among groups in their basal insulin and glucose levels and after an oral dose of glucose. Our results suggest that overfeeding or underfeeding during the suckling period affects the glucose insulin axis only temporarily and not permanently, but early overfeeding permanently enhances fatty acid synthase and lipogenic activities in adipose tissues, resulting in fatter adult rats. PMID- 11044490 TI - Expression of intrapulmonary surfactant apoprotein-A in autopsied lungs: comparative study of cases with or without pulmonary hypoplasia. AB - To investigate the functional maturity of the lungs of infants with pulmonary hypoplasia, we measured the expression of surfactant apoprotein-A (SP-A) in the autopsied lungs. Autopsied lungs were taken from 16 infants who died at birth or soon after. A lung-to-body weight ratio of less than 1.2% was defined as pulmonary hypoplasia. Eight infants were classified as belonging to the normal group, and eight as belonging to the pulmonary hypoplasia group. Many of the pulmonary hypoplasia group were complicated not only by pulmonary hypoplasia, but also by amniotic fluid volume abnormalities or an anatomical malformation. We measured the expression of SP-A immunologically using murine anti-human SP-A MAb in the autopsied lung tissue, and subjected the tissue to SP-A staining by the direct staining method. The expression of SP-A was assessed as one of four grades: -, +/-, 1+, 2+. The staining intensity of SP-A was high at 1+ or stronger in five infants of the normal group. SP-A expression was significantly reduced, however, in all infants of the pulmonary hypoplasia group except for one infant with normal amniotic fluid volume and relatively mild pulmonary hypoplasia. There was a significant negative correlation between the staining intensity of SP-A and two factors: pulmonary hypoplasia and abnormal amniotic fluid volume (p = 0.039 and p = 0.0063, respectively). In the present study, we demonstrated that SP-A expression was significantly reduced in infants with pulmonary hypoplasia. We speculate that the functional maturity of the lungs of infants with pulmonary hypoplasia is also suppressed. PMID- 11044491 TI - Leukocyte and endothelial activation in a laboratory model of extracorporeal membrane oxygenation (ECMO). AB - An inflammatory response and a capillary leak syndrome frequently develop during the treatment of neonatal respiratory failure by extracorporeal membrane oxygenation (ECMO). The present study was performed to investigate leukocyte activation and endothelial cell dysfunction that are associated with prolonged contact of blood components with synthetic surfaces. Laboratory ECMO was performed with fresh human blood at 37 degrees C for 8 h (n = 6). Leukocyte activation was measured by L-selectin (CD62L) and CD18 integrin surface expression and by neutrophil-derived elastase release. To monitor endothelial activation, endothelial cell ICAM-1 (CD54) expression was measured in cultured endothelial cells from human umbilical veins (HUVEC) after incubation with plasma from the ECMO experiments. CD18 integrin expression was found significantly up regulated on polymorphonuclear neutrophils and monocytes after 2-4 h of laboratory ECMO. L-selectin was reduced on both cell types during the total duration of the experiments. Soluble L-selectin (sCD62L) and total and differential leukocyte counts remained unchanged during the experiment. Neutrophil-derived elastase content was maximal after 8 h of ECMO. Plasma from the ECMO experiments did not induce ICAM-1 expression of cultured HUVEC. We conclude that prolonged contact with synthetic surfaces during ECMO activates phagocytes, which may contribute to the inflammatory response seen in ECMO treated patients. Activated phagocytes do not accumulate in the extracorporeal system nor release humoral factors inducing ICAM-1 expression on endothelial cells. PMID- 11044492 TI - Structural and functional features of modified heat-stable toxins produced by enteropathogenic Klebsiella cells. AB - Heat-stable enterotoxins (STs) are 18- or 19-amino acid peptides (STa or ST1) produced by enteropathogenic bacteria with small differences in their amino acid sequence and a highly conserved carboxy terminus. All STs contain a core of three disulfide bridges whose integrity is believed to be necessary for full biologic activity. We previously reported that strains of Klebsiella pneumoniae transformed by the plasmid pSLM004 produce a modified toxin not recognized by MAb raised against genuine Escherichia coli ST. Investigation of the chemical structure of the modified toxins revealed that three new toxins were present. These were purified to homogeneity by a series of sequential chromatography on reverse-phase columns using guanylate cyclase to monitor the enterotoxic activity during purification procedures. The sequence of the modified toxins was obtained by a combination of Edman degradation and mass spectrometry, showing that they are proteolytically processed forms of E. coli ST1b. In particular, toxin A-2 lacks the cysteine at position 18 and then is not able to form the disulfide bridge cysteine-10-cysteine-18. All three toxins showed the ability to stimulate guanylate cyclase and to elicit chloride secretion in Caco-2 cell monolayers mounted in Ussing chambers. Toxin A-1 and toxin B demonstrated greatly reduced immunoreactivity whereas toxin A-2 was not recognized at all in the ELISA. It is likely that the three modified toxins were generated by Klebsiella specific proteolytic processing of the original pretoxin. These results have important implications for the diagnosis and prevention of heat-stable toxin-induced diarrhea. PMID- 11044493 TI - Regulation of fatty acid transport protein and mitochondrial and peroxisomal beta oxidation gene expression by fatty acids in developing rats. AB - Regulation of genes involved in fatty acid (FA) utilization in heart and liver of weanling rats was investigated in response to variations in dietary lipid content and to changes in intracellular FA homeostasis induced by etomoxir, a blocker of FA import into mitochondria. Northern-blot analyses were performed using cDNA probes specific for FA transport protein, a cell membrane FA transporter; long chain- and medium-chain acyl-CoA dehydrogenases, which catalyze the first step of mitochondrial FA beta-oxidation; and acyl-CoA oxidase, a peroxisomal FA beta oxidation marker. High-fat feeding from postnatal d 21 to 28 resulted in a coordinate increase (58 to 136%) in mRNA abundance of all genes in heart. In liver, diet-induced changes in mitochondrial and peroxisomal beta-oxidation enzyme mRNAs (from 52 to 79%) occurred with no change in FA transport protein gene expression. In both tissues, the increases in mRNA levels went together with parallel increases in enzyme activity. Changes in FA homeostasis resulting from etomoxir administration led to a marked stimulation (76 to 180%) in cardiac expression of all genes together with parallel increases in enzyme activities. In the liver, in contrast, etomoxir stimulated the expression of acyl-CoA oxidase gene only. Feeding rats a low-fat diet containing 0.5% clofibrate, a ligand of peroxisome proliferator-activated receptor alpha, resulted in similar inductions of beta-oxidation enzyme genes in both tissues, whereas up-regulation of FA transport protein gene was restricted to heart. Altogether, these data suggest that changes in FA homeostasis in immature organs resulting either from high-fat diet or beta-oxidation blockade can efficiently be transduced to the level of gene expression, resulting in tissue-specific adaptations in various FA-using enzymes and proteins. PMID- 11044494 TI - Evidence for change of 11beta-hydroxysteroid dehydrogenase activity during infancy and childhood. AB - The conversion of cortisol (F) to cortisone (E) is catalyzed by 11beta hydroxysteroid dehydrogenase type 2. The present study was designed to investigate the changes of F and E plasma concentration as an indirect measurement of 11beta-hydroxysteroid dehydrogenase activity in infancy and childhood. Plasma samples were obtained from 262 healthy children and adolescents aged 1 d to 18 y. Plasma F and E were measured, using specific radioimmunoassays after extraction and automated Sephadex LH 20 chromatography. The F/E ratio was calculated to assess 11beta-hydroxysteroid dehydrogenase activity. During the first year of life, plasma F levels rose significantly (r(2) = 0,24; p = 0.01), and thereafter no further increase was seen until adulthood (r(2) = 0.01; p = 0.86). In contrast, plasma E significantly decreased during the first year of life (r(2) = -0.35; p<0.001) and stayed unchanged thereafter (r(2) = 0.02; p = 0.81). As a consequence, the F/E ratio rose significantly during the first year (r(2) = 0.67; p<0.001) but did not change afterward (r(2) = 0.001; p = 0.99). During the first year of life, there is a change from the predominance of E, with low mineralocorticoid receptor affinity, to F, with high mineralocorticoid receptor affinity. This shift corresponds to the declining plasma concentrations of aldosterone during infancy. The changes may indicate a not yet recognized, significant change of 11beta-hydroxysteroid dehydrogenase isoenzyme activity or alterations in the secretion of F and E, which may be of relevance for the development of arterial blood pressure in infancy. PMID- 11044495 TI - Steady state is not achieved for most plasma amino acids during 12 hours of fasting in the neonatal piglet. AB - Kinetics studies in neonates are important to establish the requirement for amino acids and to understand the mechanisms of normal and altered metabolism. During kinetics experiments, plasma amino acid concentrations should be in steady state. Our objective was to determine whether 12 h of fasting, after parenteral or enteral feeding, resulted in a steady state in concentrations of amino acids. Two day-old piglets were implanted with catheters (d 0), and randomly assigned to either intragastric (i.g., n = 6) or i.v. (n = 6) feeding. On d 5, piglets were fasted for 12 h. During the first 2 h, plasma concentrations of almost all amino acids declined except asparagine (i.g. and i.v.), tyrosine (i.v.), and glycine (i.v.), which increased. Only i.g. glycine did not change. Between 2 and 12 h, the only indispensable amino acids that did not change were phenylalanine (i.v.) and histidine (i.g. and i.v.). The branched-chain amino acids increased during this period (i.v. and i.g.). The greatest change was tyrosine, increasing 13% (i.v.) and 32% (i.g.) per hour. After 12 h of refeeding, glycine, serine, threonine, and asparagine concentrations were lower than baseline (p<0.05) in the i.v. group. In i.g. fed piglets, only threonine remained below baseline (p<0.05), and arginine was greater than baseline (p<0.05). Differences between i.v. and i.g. may be the result of impaired small intestinal metabolism secondary to parenteral feeding. In neonatal pigs, most plasma amino acids were unstable during 12 h of fasting. Thus, kinetics studies that require a steady state must be conducted in the fed state. PMID- 11044496 TI - Approaching the adnexal mass in the new millennium. AB - Adnexal masses are common dilemmas faced by practicing gynecologists. They affect women from before birth throughout life, yet considerable disagreement exists regarding their optimal management. Traditional management focused on avoiding undertreatment of a potentially malignant process. Advances in detection, diagnosis, and minimally invasive management make it necessary to review this practice to avoid unnecessary morbidity and mortality. The literature emphasizes a minimally invasive approach to the treatment of benign lesions without sacrificing the principles of oncologic surgery. PMID- 11044497 TI - Sources of error when tracking irrigation fluids during hysteroscopic procedures. AB - STUDY OBJECTIVE: To evaluate the ability of circulating nurses to estimate input and outgo of irrigating fluids used during hysteroscopic procedures in a hospital operating room. DESIGN: Simulation of intraoperative measurements (Canadian Task Force classification II-1). SETTING: Operating room. Intervention. Circulating nurses estimated fluid volumes under circumstances simulating actual conditions of hysteroscopic ablative or resection procedures. MEASUREMENTS AND MAIN RESULTS: Three-liter glycine irrigation bags were overfilled by an average of 2.8% (62-125 ml). Estimates of fluid remaining in partially emptied bags were in error by an average ranging from 4% to 50%/bag (largest error 10-55%, 157-401 ml). Estimates of fluid in kick buckets were in error by an average of 10% to 39% (largest error 22-66%, 232-903 ml). Visual estimates of fluid on the operating room floor were in error by an average of 56% to 67% (largest error 65-81%, 182-840 ml). Estimates of fluid in suction canisters were consistent among nurses. The accuracy of measurements for partially filled suction canisters primarily depended on the accuracy of canister calibration. Volume contained in cascaded suction canisters from an actual surgical procedure was grossly different from rated capacity. CONCLUSION: Accurate tracking of irrigation fluid during hysteroscopic procedures is difficult. Even with a mechanical measuring system, fluid lost on the floor can introduce sizable errors. Estimation errors can easily and quickly accumulate to clinically significant volumes. Use of an automated mechanical fluid-tracking system with devices to capture fluid lost from the surgical field is recommended. PMID- 11044498 TI - An evidence-based medicine approach to the treatment of endometriosis-associated chronic pelvic pain: placebo-controlled studies. AB - OBJECTIVE: Use an evidence-based medicine (EBM) approach to evaluate the evidence regarding efficacy of treatment of endometriosis-associated chronic pelvic pain (CPP) in placebo-controlled randomized clinical trials (RCT). DESIGN: Review of six randomized, controlled trials (Canadian Task Force classification I). SETTING: University of Rochester School of Medicine and Dentistry. Patients. Three hundred eighty-one women with endometriosis enrolled in placebo-controlled randomized clinical trials. Intervention. A MEDLINE search of published medical articles from January 1976, to January 1998. MEASUREMENTS AND MAIN RESULTS: Six placebo-controlled randomized clinical trials were found that addressed the treatment of pelvic pain associated with endometriosis and met validity criteria; one was a study of surgical treatment, two of medical therapies, and three of combined surgical and medical treatments. They clearly show that laparoscopic surgery and medical treatment with medroxyprogesterone acetate, danazol, or nafarelin are more effective than placebo. Evidence for efficacy of leuprolide acetate is weaker. At 6 months, absolute decreases in pain scores are quite similar with surgical or medical treatment. Medical therapy after surgical treatment significantly reduced pain, but six months after it was stopped there was no difference between women treated and not treated postoperatively. CONCLUSIONS: Although either surgical or medical treatment of endometriosis in women with CPP is clearly indicated, pain relief of 6 or more months' duration can be expected in only 40 to 70% of women with endometriosis-associated CPP. PMID- 11044499 TI - One-year results of the vesta system for endometrial ablation. AB - STUDY OBJECTIVE: To compare a distensible multielectrode balloon for endometrial ablation with electrosurgical ablation performed by a combined resection coagulation technique. DESIGN: Randomized, prospective trial (Canadian Task Force classification I). Setting. Eight centers. PATIENTS: Women with menorrhagia validated with a standardized pictorial blood loss assessment chart (PBAC), without intracavitary organic uterine disease, who failed or poorly tolerated medical therapy. Intervention. Results in 122 patients treated by Vesta and 112 treated surgically, evaluable at 1 year, were compared, with success defined as monthly blood loss of less than 80 ml and avoidance of additional therapy. MEASUREMENTS AND MAIN RESULTS: Pretreatment PBAC scores for patients treated by Vesta and resection or rollerball were 535+/-612 and 445 +/- 313, respectively; at 1 year they were 18+/-37 and 28+/-60, respectively. With PBAC below 75 as the definition of success, 86.9% of Vesta-treated patients were successful compared with 83.0% treated by rollerball or resection. Total amenorrhea, defined as no visible bleeding and no use of protective products, was 31.1% and 34. 8%, respectively. None of the outcome comparisons between treatments showed statistical difference. Complications in both groups were few and minor. Most (86.6%) Vesta procedures were carried out with paracervical block with or without intravenous sedation in an office or outpatient setting, compared with 79.7% epidural or general anesthesia for rollerball or resection. CONCLUSION: The Vesta system of endometrial ablation is equally effective and safe as classic resectoscopic methods. Potential advantages include avoidance of fluid and electrolyte disturbance associated with intravasation of distending media, and ability to perform the procedure under local anesthesia in an office setting with less total operating time. PMID- 11044500 TI - Preliminary experience with the VersaPoint bipolar resectoscope using a vaporizing electrode in a saline distending medium. AB - OBJECTIVE: To evaluate a new bipolar resectoscope that uses physiologic saline as a distending medium. DESIGN: Clinic-based, prospective, non-randomized study (Canadian Task Force classification II-2). SETTING: A free-standing ambulatory surgical facility. Patients. Eleven women with menorrhagia or menometrorrhagia and four with menorrhagia and infertility. INTERVENTION: Hysteroscopic removal of submucosal myomas with concomitant endometrial ablation in seven patients. MEASUREMENTS AND MAIN RESULTS: Complete removal of the submucosal myoma was achieved in all 15 patients. Eight patients had a European Society of Gynecologic Endoscopy type II myoma, five had a type I, and two had a type zero. Five patients had multiple myomas. There were no complications. CONCLUSIONS: The bipolar resectoscope is effective in the removal of submucosal myomas and in this series allowed completion of several cases that probably could not have been done with a traditional monopolar resectoscope. PMID- 11044501 TI - Litigation of laparoscopic major vessel injuries in Canada. AB - This study is a review of conditions and circumstances associated with 15 injuries to great vessels during laparoscopic surgery. Thirteen cases were litigated in Canada and two occurred in the author's operating room. Body habitus may have been an underlying factor in the injuries. Most injuries were entry related and independent on complexity of surgery. One was caused by the Veress needle and one by a secondary trocar, and three occurred during dissection of adhesions. Ten were primary trocar injuries, 9 after pneumoperitoneum and 1 at direct trocar insertion. Of these, five were caused by reusable and five by disposable trocars with so-called safety shields. In 6 of 15 cases recognition of the injury was delayed; 5 in the recovery room. Eleven women had uncomplicated recovery. Of 13 litigated cases, 8 (62%) resulted in settlement. PMID- 11044502 TI - Laparoscopic-assisted vaginal myomectomy. AB - STUDY OBJECTIVE: To evaluate the safety and efficacy of combined laparoscopic and vaginal approach in dealing with uterine myomas. DESIGN: Retrospective case study (Canadian Task Force classification II-2). SETTING: Tertiary care major teaching hospital. PATIENTS: Thirty-one women with symptomatic fundal and/or posterior wall uterine myomas. INTERVENTION: Laparoscopic-assisted vaginal myomectomy performed by one of the authors from July 1996 to December 1998. MEASUREMENTS AND MAIN RESULTS: Mean +/- SD operating time, blood loss, and length of hospital stay were 79.19+/-18.31 minutes, 150.00+/- 103.28 ml, and 3.10+/-0.75 days, respectively. No patients developed serious complications, and only two minor complications occurred. CONCLUSION: After laparoscopic inspection and location of uterine myomas, dealing with posterior and fundal uterine myomas by the vaginal route makes hemostasis and uterine repair easier than by purely laparoscopic approach. PMID- 11044503 TI - Frequency of lower urinary tract injury at laparoscopic burch and paravaginal repair. AB - STUDY OBJECTIVE: To estimate the rate of injury to the lower urinary tract during laparoscopic Burch urethropexy and/or paravaginal repair. DESIGN: Retrospective analysis over 30 consecutive months (Canadian Task Force classification II-2). Setting. Community hospital. PATIENTS: One hundred seventy-one consecutive patients. INTERVENTION: Laparoscopic Burch urethropexy and/or paravaginal repair. MEASUREMENTS AND MAIN RESULTS: All patients had intraoperative transurethral videocystoscopy performed with intravenous injection of indigo carmine dye to assess potential injury to bladder or ureter. Four women (2.3%, CI -0.71-0.03) had injury to the lower urinary tract. All four injuries were cystotomies, two in women with previous open retropubic urethropexy. No ureteral ligation or intravesical placement of suture was diagnosed. CONCLUSION: Despite most patients having both Burch urethropexy and paravaginal repair, the lower urinary tract injury rate of 2.3% is much lower than the reported 10% for patients having Burch urethropexy alone performed by laparotomy. Reported benefits of laparoscopy including less blood loss and better visualization may explain this result. PMID- 11044504 TI - Reliability of frozen section of uterine curettings in evaluation of possible ectopic pregnancy. AB - STUDY OBJECTIVE: To assess the accuracy of frozen section evaluation of uterine curettings to look for chorionic villi and rule out ectopic pregnancy before laparoscopy. DESIGN: Retrospective review (Canadian Task Force classification III). SETTING: Northeastern medical school-affiliated teaching hospital. PATIENTS: Thirty-six women. Intervention. Dilatation and curettage, laparoscopy, and laparotomy. MEASUREMENTS AND MAIN RESULTS: Of 36 cases, 13 showed evidence of intrauterine pregnancy, either chorionic villi or implantation site, on final permanent sections. Five false negatives were identified in which no villi were identified on frozen section, although villi or evidence of an implantation site was noted on permanent sections (sensitivity 62%, 95% CI 0.32, 0.86). CONCLUSION: Frozen section evaluation of uterine curettings can produce false negative diagnoses, and this should be considered in the operative planning of women with suspected ectopic pregnancy. PMID- 11044505 TI - Decrease in the number of abdominal hysterectomies after introduction of laparoscopic hysterectomy. AB - STUDY OBJECTIVE: To compare two 3-year periods before and after laparoscopic hysterectomy was introduced into our resident training program. DESIGN: Retrospective analysis (Canadian Task Force classification II-2). SETTING: Teaching hospital in the Netherlands. PATIENTS: Women undergoing hysterectomy from 1992 to 1994 and 1995 to 1997. INTERVENTION: Abdominal, vaginal, and laparoscopic hysterectomies. MEASUREMENTS AND MAIN RESULTS: Laparoscopic hysterectomy significantly (p<0.002) reduced the number of abdominal hysterectomies. CONCLUSION: To reduce the number of abdominal hysterectomies, it is essential that the laparoscopic procedure be taught to residents. PMID- 11044506 TI - Continuous-flow vaginoscopy in children and adolescents. AB - STUDY OBJECTIVE: To assess the role of continuous-flow vaginoscopy in the management of gynecologic problems in pediatric and adolescent patients. DESIGN: Prospective, observational study (Canadian Task Force classification II-3). Setting. University-affiliated teaching hospital. PATIENTS: Twenty-two consecutive children and adolescents evaluated for vulvovaginitis (15), vaginal trauma (4), bleeding (2), and genital malformation (1). INTERVENTION: Continuous flow vaginoscopy with a 4-mm hysteroscope under general anesthesia. MEASUREMENTS AND MAIN RESULTS: Vaginal walls, fornices, and cervices were well visualized in all patients. No pathologic findings were found in 16, a foreign body was present in 3, and vaginal lacerations in 3. Foreign material was removed with long straight forceps, bleeding spots were coagulated, and lacerations sutured. No complications occurred. The patients were discharged 4 to 24 hours after the procedure. CONCLUSION: Diagnosis of gynecologic problems in children and adolescents should include vaginoscopy. Continuous-flow vaginoscopy is quick and easy to perform in these patients. PMID- 11044507 TI - Complications of laparoscopic surgery for benign ovarian cysts. AB - STUDY OBJECTIVE: To assess complications of laparoscopic surgery in the management of ovarian cysts. DESIGN: Prospective observational study (Canadian Task Force classification II-2). SETTING: University-affiliated hospital. PATIENTS: Consecutive patients (513) undergoing laparoscopic surgery for ovarian cysts not suspected to be malignant. INTERVENTION: Laparoscopic surgery. MEASUREMENTS AND MAIN RESULTS: A total of 587 ovarian cysts were removed from 513 women. Conversion to laparotomy was necessary in five cases (<1%). Mean +/- SD cyst diameter was 5.5+/-2.9 cm, with endometriomas (44. 5%) and dermoids (24.3%) being the two most common pathologies; 6.6% were functional. Mean +/- SD operating time was 69+/-31 minutes, and hospital stay and postoperative convalescence was 2.6+/-1.5 and 14.3 +/-9.6 days, respectively. The overall complication rate was 13.3%. Major complications occurred in three patients (0.6%): one small bowel injury and two ureter injuries. Cannula site complications were five inferior epigastric vessel injuries and four incisional hernias at the 10-mm lateral port site. CONCLUSION: Laparoscopic ovarian surgery was associated with 13.3% complications, with 0.6% being major. Careful patient selection and proper surgical training are critical to ensure safe performance of laparoscopy. PMID- 11044508 TI - Transvaginal videopelviscopy, a new technique for assessing pelvic cysts. AB - We evaluated a new technique, transvaginal videopelviscopy, of inspecting the internal architecture of pelvic cysts and verifying the possibility of performing intracystic biopsy through the posterior fornix. The procedure was performed in 33 women with pelvic cysts just before operative videolaparoscopy. A 1.2-mm scope was introduced through the pouch of Douglas under transvaginal ultrasound guidance. The inner surface of adnexal cysts was visualized and attempts were made to secure biopsy tissue. Good images of the inner wall of the lesion were obtained in 26 patients (77.8%). Specimens for histopathology were obtained in 27 women (81. 8%) and were sufficient for classifying the tumor as neoplastic or nonneoplastic. On histopathologic analysis, intracystic biopsy tissue and specimens obtained by laparoscopy were concordant in 25 (92.6%) of 27 patients. No complications occurred. Transvaginal videopelviscopy is an effective procedure for assessing pelvic cysts. PMID- 11044509 TI - Ovarioscopy, a technique to determine the nature of cystic ovarian tumors. AB - STUDY OBJECTIVE: To test the value of ovarioscopy as an additional step in the diagnostic work-up of probably benign cystic ovarian tumors before laparoscopic intervention, and to compare its diagnostic accuracy with that of transvaginal ultrasonography (TVUS) and tumor markers. DESIGN: Cross-sectional study (Canadian Task Force classification II-2). SETTING: Gynecologic endoscopy unit at a university hospital. PATIENTS: Sixty-eight women with unilateral or bilateral ovarian cystic swellings without clinical, sonographic, or laparoscopic suspicion of malignancy. INTERVENTION: Preoperatively, TVUS and tumor markers were estimated. Intraoperative endocystic ovarioscopic visualization ovarioscopy and ovarioscopy-guided biopsy were done before laparoscopy. MEASUREMENTS AND MAIN RESULTS: Ovarioscopy had the highest specificity for detecting benign ovarian cysts (98%) compared with (72.6%) and (72%) for tumor markers and TVUS, respectively. Its positive predictive value was 50% compared with 5% and 6% for tumor markers and TVUS, respectively. Its findings agreed with the histopathologic diagnosis in 39 patients (57%, p = 0.000, k = 0.85). CONCLUSION: Ovarioscopy is a simple, rapid maneuver that should precede laparoscopic ovarian cystectomy. It is superior to tumor markers and TVUS for predicting the benign nature of ovarian cysts. PMID- 11044510 TI - Current state of office laparoscopic surgery. AB - STUDY OBJECTIVE: To summarize office laparoscopic surgery practice profiles with emphasis on microlaparoscopy. DESIGN: Survey (Canadian Task Force classification III). SETTING: General gynecology and reproductive endocrinology laparoscopic surgery practices in the United States and abroad. INTERVENTION: Surveys were distributed to members of the American Association of Gynecologic Laparoscopists during 1999. PARTICIPANTS: Of 6200 members to whom surveys mailed, 1504 (24.3%) responded. MEASUREMENTS AND MAIN RESULTS: Of these respondents, 114 (7.6%) perform office laparoscopy and 187 (12.4%) microlaparoscopy. CONCLUSION: Clinicians who perform office laparoscopy are more likely to perform microlaparoscopy than macrolaparoscopy. The 2-mm microlaparoscope is preferred by most physicians. PMID- 11044512 TI - On-Q system for managing trocar site pain after operative laparoscopy. AB - The On-Q system provides long-term pain relief with continuous infusion of a local anesthetic directly into the trocar site after operative laparoscopy. An elastometric pump filled with bupivacaine HCl and a catheter, inserted by the surgeon, delivers 48 hours of anesthesia (100 ml) at a rate of 2 ml/hour. The pump is secured to the outer surgical dressing or to the patient's clothing with tape or an E clip provided in the package. The system can be removed by the patient when the infusion is complete. PMID- 11044511 TI - Laparoscopic plication and suspension of the round ligament for chronic pelvic pain and dyspareunia. AB - STUDY OBJECTIVE: To describe a technique of uterine suspension using round ligaments to relieve pain in selected patients with various degrees of uterine retroversion. DESIGN: Prospective clinical study (Canadian Task Force classification II-2). SETTING: University-based center for reproductive medicine. PATIENTS: Thirty women who underwent laparoscopy for investigation of chronic pelvic pain (CPP) and dyspareunia. INTERVENTION: Round ligaments were plicated using a modification of intracorporeal knot tying during laparoscopy. MEASUREMENTS AND MAIN RESULTS: The extent of plication was planned to elevate the uterus and bring it minimally forward. Mean +/- SD operating time was 14+/-4 minutes. Pain scores before and after surgery were 4.5+/-1.0 and 1.6+/-0.6, respectively (p<0.001). There were no complications during or after surgery. Only three women had minimal dyspareunia postoperatively, although they had anteverted uteri. One patient had severe dyspareunia that developed 7 months after surgery and continued through the second year of follow-up. Nineteen women with anteverted uteri were free of dyspareunia after 2 years. CONCLUSION: Round ligament plication is safe and effective in patients with retroverted uteri and dyspareunia or CPP. PMID- 11044513 TI - Interrupted laparoscopic suture secured with the needle-through-the-sliding-loop knot. AB - A special needle holder with a step and knot-forming stand was designed to permit wrapping sutures into distinctive sliding loops around the needle holder, outside the abdomen. The loaded instrument was introduced into the abdomen and the appropriate stitch was made. The suture next to the needle was grasped with the needle holder and pulled through the sliding loops(s) to establish a knot. When the needle was passed through a simple noose the needle-through-the-noose knot was created. This knot maintained tension on captured tissues, but it required additional half-hitches to secure. When the needle was passed through rolling hitch loops it established a knot that was secure in simple applications, but required one additional half-hitch to secure in vulnerable applications involving slippery sutures or tissue purchases subjected to disruptive tension. The procedures were first practiced on inanimate models using a laparoscopy training simulator and then performed successfully over 20 times in 11 patients who had laparoscopic myomectomy or hysterectomy. PMID- 11044514 TI - Transvaginal hydrolaparoscopy, a new technique for pelvic assessment. AB - Transvaginal hydrolaparoscopy is based on classic culdoscopy. With alterations in equipment and method, the procedure holds promise for evaluation of pelvic pathology. We performed transvaginal hydrolaparoscopy in the operating room just before operative laparoscopy in 15 patients, to evaluate the feasibility of this procedure. Excellent images of the cul-de-sac, fimbriae, and caudal surface of the uterus, ovaries, and pelvic sidewall were obtained. We believe this is a practical and convenient office diagnostic procedure. PMID- 11044515 TI - Urinary tract injuries during advanced gynecologic laparoscopy. AB - Urinary tract injuries are important complications of laparoscopic surgery. The intraoperative diagnosis may be delayed, resulting in severe clinical complications, such as fistulas, in the immediate and late postoperative periods. A review of 776 endoscopic procedures revealed 6 urinary tract injuries and postoperative complications during laparoscopy. We believe that surgical experience, intraoperative diagnosis, immediate repair of the lesion, and close follow-up are the main factors contributing to decreased morbidity associated with these injuries. PMID- 11044516 TI - Disseminated intravascular coagulation as a complication of intrauterine balloon tamponade for posthysteroscopic acute uterine bleeding. AB - Although there is anecdotal evidence of balloon tamponade for posthysteroscopic acute uterine bleeding, a literature review disclosed no articles describing complications of the technique. In our patient, balloon tamponade for acute posthysteroscopic bleeding resulted in disseminated intravascular coagulation. In a second woman the procedure was successful. PMID- 11044517 TI - The reality of the healthcare marketplace and the medical school curriculum. PMID- 11044518 TI - Video-assisted endoscopic thyroidectomy. AB - BACKGROUND: Several experimental and clinical reports concerning endoscopic parathyroid surgery have appeared. However, reports concerning minimally invasive surgery for thyroid remains rare. Herein we present a new method, called video assisted endoscopic thyroidectomy (VAET), for the management of various benign thyroid diseases. METHODS: In all, 16 consecutive patients who underwent VAET for benign thyroid diseases were retrospectively studied. The study group included nodular hyperplasia in 8 patients, follicular adenoma in 6, and Hurthle's tumor and simple cyst in 1 each. A 2 to 3 cm transverse incision was made on the suprasternal notch. The wound was deepened to expose the underlying trachea from which the plane of the thyroid fascia was accessed directly, and the working space was established with lifting method using conventional instrument. All surgical procedures could be manipulated and monitored under laparoscopy without gas insufflation. The ultrasonically activated scalpel was the principal instrument used for VAET. RESULTS: All 16 patients underwent VAET successfully without conversion to open thyroidectomy. The surgical procedures included lobectomy in 13 and extirpation in 3. The operation time ranged from 28 minutes to 5 hours (mean 1 hour, 42 minutes). For the 5 most recent cases, lobectomy took an average of 2 hours, whereas extirpation less than 40 minutes. The tumor size ranged from 3.5 cm to 8.0 cm (mean 5.8 cm). There were no surgical complications. All patients but 1 were discharged on postoperative day 2. During follow-up, all patients demonstrated euthyroid function and satisfactory cosmetic results. CONCLUSIONS: VAET emerges as a promising minimally invasive surgical technique replacing conventional thyroidectomy for benign thyroid diseases in selected cases, with the advantage of satisfactory cosmetic results. PMID- 11044519 TI - A history of the dissolution of retained choledocholithiasis. AB - BACKGROUND: Common duct calculi retained after gallbladder surgery continue to present a clinical challenge especially in the era of minimally invasive surgery. This review examines the strategy of dissolution therapy used throughout the history of biliary tract surgery and its use to the modern surgeon. DATA SOURCES: Original journal articles and reviews were identified using standard surgical textbooks and MEDLINE. Keywords for searching included choledocholithiasis, dissolution, mono-octanoin, common duct stones, MTBE, cholic acid, and gallstones. CONCLUSIONS: Dissolution therapy used initially as an alternative to open surgery is now used more effectively as an adjunct to laparoscopic or endoscopic biliary tract surgery. The current review demonstrates a majority of patients with retained choledocholithiasis respond to dissolution and can be safely managed without choledochotomy. PMID- 11044520 TI - Gallstone ileus. PMID- 11044521 TI - Axillofemoral bypass for aortoiliac occlusive disease. AB - BACKGROUND: Although aortoiliac disease remains a common cause of lower extremity ischemia, the efficacy of axillofemoral bypass in this setting remains controversial. This report summarizes our institutional experience with axillofemoral bypass. METHODS: A retrospective review of consecutive axillofemoral bypass grafts was performed at a single institution between 1984 and 1997. Only patients presenting with chronic aortoiliac occlusive disease were included. Patient demographics, risk factors, indications for surgery and outcomes were recorded. Survival curves for primary patency were plotted using the Kaplan-Meier method according to the standards set by the Society of Vascular Surgery-International Society for Cardiovascular Surgery. Comparisons between groups were made using the log rank method. Statistical significance was assumed at P values <0.05. RESULTS: Sixty patients underwent axillofemoral bypass grafting of which 53 were bifemoral and 8 unifemoral. Forty-seven procedures were performed for limb salvage. Primary patency rates at 1, 3, and 5 years were 86%, 72%, and 63%, respectively. Thirty-day mortality rate was 4.9%. Sixty percent of graft occlusions occurred in the femorofemoral limb with continued patency of the axillofemoral limb. Risk factors, type of procedure and superficial femoral artery patency had no statistically significant effect on long-term patency. CONCLUSIONS: In the setting of diffuse, chronic aortoiliac occlusive disease, long-term patency rates of axillofemoral grafts approach those of aortobifemoral bypass and exceed those quoted for percutaneous transluminal angioplasty, with results that are highly reproducible. Axillofemoral bypass is an excellent option in those patients at prohibitive risk for direct aortic reconstruction or those with limited life expectancy. PMID- 11044522 TI - A prospective study of seeding of the skin after core biopsy of the breast. AB - BACKGROUND: The number of core biopsies done for breast abnormalities is increasing. The risk of skin seeding resulting from core biopsy is unknown. METHODS: Consecutive patients diagnosed with breast cancer were studied. The skin and subcutaneous fat surrounding the site of core needle penetration were excised and studied by routine histologic staining. Findings were correlated with other clinical variables. RESULTS: Eighty-nine consecutive patients were studied. Thirty-one had stereotactic core biopsies, 23 had vacuum-assisted biopsy, 8 had multiple-puncture biopsy, and 58 had ultrasound-guided core biopsy. Two patients who were biopsied using multiple-puncture biopsy were found to have nests of cancer cells in the dermis. One of these patients had recurrence in the skin biopsy site at 34 months. CONCLUSION: Skin seeding may be important in light of increasing use of image-directed biopsy, and particularly for cases in which the biopsy puncture site is outside the index quadrant and in which no radiation is anticipated. PMID- 11044523 TI - Effect of hematocrit on regional oxygen delivery and extraction in an adult respiratory distress syndrome animal model. AB - BACKGROUND: The purpose of this prospective, randomized, controlled study was to investigate the effects of hematocrit (Hct) on regional oxygen delivery and extraction following induction of adult respiratory distress syndrome (ARDS) in an animal model. METHODS: Animals were instrumented to monitor central venous pressure (CVP), systemic mean arterial pressure (MAP), pulmonary artery occlusion pressure (PAOP), and cardiac output (CO) and to measure blood flow in the renal, hepatic, and superior mesenteric arteries and portal vein. ARDS was induced, positive end expiratory pressure (PEEP) applied and CO was maximized with volume loading and epinephrine infusion. Data were acquired at baseline (BL) and at Hct levels ranging from 25% to 50%. RESULTS: Systemic DO(2) increased steadily and significantly with increased Hct. Systemic O(2) extraction ratio (O(2)ER) decreased significantly with increasing Hct until a threshold value of 40%, after which further increases in Hct did not cause a statistically significant decrease in O(2)ER. Similarly, renal and hepatic DO(2) increased and O(2)ER decreased in a statistical significant manner with transfusions up to a Hct of 35%. In the splanchnic circulation blood transfusions did not cause any statistically significant increase in DO(2), and O(2)ER showed no decrease after an Hct of 35%. Systemic, renal, hepatic, and splanchnic VO(2) were not affected by changes in Hct. Blood viscosity decreased from a baseline value of 2.9+/-0.2 centipoise at a Hct of 38% to 2.3+/-0.1 centipoise at a Hct of 25% (P<0.05). Viscosity increased progressively with increasing hematocrits and reached the value of 4.2+/-0.2 centipoise at an Hct of 50% (P<0.05 versus Hct 30%, 35%, 40%, 45%). CONCLUSIONS: Based on the results of this non-supply-dependent animal model we conclude that a progressive increase in Hct up to 40% causes a corresponding increase in systemic DO(2) associated with a decrease in O(2)ER. However, there is no improvement in renal, hepatic, and splanchnic DO(2) and O(2)ER after a threshold Hct of 35%. All other factors being the same, an Hct greater than 35% may in fact cause a decrease in blood flow rate and change in blood flow characteristics as a consequence of increased blood kinematic viscosity, which may alter and compromise cellular oxygen transfer. PMID- 11044524 TI - Pancreaticojejunal stent migration resulting in "bezoar ileus". PMID- 11044525 TI - Pancreatic anastomotic failure after pancreaticoduodenectomy. AB - BACKGROUND: Pancreatic anastomotic failure has historically been regarded as one of the most feared complications after pancreaticoduodenectomy. METHODS: We reviewed our recent experience (59 cases), March 1994 to December 1998, with pancreaticoduodenectomy and compared preoperative and intraoperative characteristics as well as outcomes in those patients who experienced (n = 10) versus those who did not experience a postoperative pancreatic leak (n = 49). Information was retrospectively collected from hospital records, office records, and interviews with patients. RESULTS: The clinical leak rate in this series was 8.5%. There were no significant differences in preoperative or intraoperative characteristics comparing those with versus those without a postoperative pancreatic leak. Only 1 of 10 patients with a postoperative pancreatic leak required reoperation to manage the leak. Those with a pancreatic leak had more other postoperative complications (median 2 versus 0 complications per patient, P = 0.01) and longer hospital duration compared with those without a leak (median 13 versus 23 days, P<0.01). Overall mortality in the series was 3.4%; no mortalities occurred as a result of a pancreatic leak. CONCLUSIONS: In the 1990s pancreatic anastomotic leak remains a potentially lethal problem after pancreaticoduodenectomy. Pancreatic leakage after pancreaticoduodenectomy is associated with other postoperative complications and a longer hospital stay. PMID- 11044526 TI - Whole gut washout ameliorates the progression of acute experimental pancreatitis. AB - BACKGROUND: Septic complications are mainly responsible for deterioration of a patient with acute pancreatitis. Intestinal tract is accepted as the main source of pancreatic or peripancreatic infection. MATERIAL AND METHODS: Acute pancreatitis was induced in 40 Sprague-Dawley rats by ligation of the main biliopancreatic duct. Animals were divided into two groups. The first group of animals (n = 20) received high volume polyethylene glycol-3500 (GoLYTELY) for 6 hours through a silastic catheter introduced into the proximal part of the jejunum from a puncture gastrostomy during the initial laparotomy. The second group animals (n = 20) did not receive any treatment. Half of the animals from each group were sacrificed 72 hours later and tissue samples were taken from mesenteric lymph nodes, pancreas, spleen, and liver for bacteriologic cultures. Cecum cultures were also prepared. Blood samples at 72 hours were obtained for the measurement of amylase, lactic dehydrogenase (LDH), lactic acid, alanine aminotransferase (ALT), glucose, calcium, arterial pH, base excess, partial oxygen pressure, bicarbonate, leucocyte count, and hematocrit levels. The pancreas was examined histopathologically. The remaining half of the animals from each group were allowed to survive until death. RESULTS: The levels of amylase, LDH, ALT, lactic acid, pH, pO(2), bicarbonate and base excess for the rats in group I were significantly lower when compared with the rats in group II (P<0.05). Positive mesenteric lymph node cultures were detected in 30% of group I animals whereas they were positive in 90% of group II animals (P = 0.0198). Distant organ cultures were positive in 8 animals (liver 5, spleen 2, pancreas 1) in group II, whereas only one positive distant organ culture (liver) was established in group I (P>0.05). Histopathological scoring observed in the pancreas were less severe for the rats in group I when compared with the rats in group II (P = 0.012). The rats in group I survived longer than the rats in group II (median survival 6.8 days versus 17.3 days, P<0.001). CONCLUSIONS: Whole gut washout with high-volume polyethylene glycol in pancreatitis reduced the blood levels of enzymes and increased the survival. Whole gut washout for acute pancreatitis appears effective to ameliorate the prognostic factors in blood and this modality may be a promising treatment method in acute pancreatitis. PMID- 11044527 TI - Incarcerated Meckel's diverticulum in a Spigelian hernia. PMID- 11044528 TI - Long-term benefit of extended lymphadenectomy with gastrectomy in distally located early gastric carcinoma. AB - BACKGROUND: Extended lymphadenectomy performed with gastrectomy has been reported to prolong survival of patients with early gastric cancer. However, some authors question the value of extensive lymphadenectomy in these patients, especially since much recent discussion of patient quality of life after gastrectomy has favored less invasive operations. METHODS: We retrospectively analyzed 485 patients who had undergone gastrectomy for early cancer in order to evaluate the effect of extended versus limited lymphadenectomy on postoperative survival. Various prognostic factors were examined for patients whose tumors were located in the distal third of the stomach. RESULTS: Although extended radical lymphadenectomy did not prolong postoperative survival when early gastric cancer was located in the middle or proximal third of the stomach, it did when the tumor occupied the distal third. CONCLUSIONS: Performance of extended radical lymphadenectomy was a significant prognostic factor for early gastric cancer patients when tumors were located in the distal third of the stomach. PMID- 11044529 TI - The clinical role of noncontrast helical computed tomography in the diagnosis of acute appendicitis. AB - BACKGROUND: The accuracy of noncontrast helical computed tomography (CT) for appendicitis has recently been demonstrated. What is its clinical utility? METHODS: This was a retrospective review of 443 consecutive community hospital patients evaluated for acute appendicitis over an 18-month period using limited pelvic CT scan or clinical acumen alone. RESULTS: Appendicitis was pathologically proven in 158 patients. The negative appendectomy rate was 5.4%. The best radiological indicators for a positive CT for appendicitis were pericecal inflammation (88%) and appendicolith(57%). Appendiceal CT was found to have a 92% sensitivity, 99.6% specificity, and a 97.5% accuracy. There were 260 patients who had a negative CT; 243 of these were sent home. Alternative diagnoses were identified in 22% of patients. CONCLUSIONS: The liberal use of noncontrast helical CT results in a low negative appendectomy rate and a high degree of confidence that a negative CT will allow patients to be sent home safely. PMID- 11044530 TI - Aneurysmosis presenting as acute limb ischemia. PMID- 11044531 TI - A comparison of the prognostic significance of tumor diameter, length, width, thickness, area, volume, and clinicopathological features of oral tongue carcinoma. AB - BACKGROUND: The present study aims at evaluation of the prognostic value of tumor size including diameter, length, thickness, width, area, and volume in the prediction of nodal metastasis, local recurrence, and survival of oral tongue carcinoma. The results will have important implications for the management of patients. METHODS: Eighty-five glossectomy specimens of oral tongue carcinoma were serially sectioned in 3 mm thickness for the tumor size evaluation with computer image analyzer. RESULTS: Among all the tumor size parameters being evaluated, tumor thickness was the only significant factor for the prediction of local recurrence, nodal metastasis, and survival. With the use of 3 mm and 9 mm division, tumor of up to 3 mm thickness has 10% nodal metastasis, 0% local recurrence, and 100% 5-year actuarial disease-free survival; tumor thickness of more than 3 mm and up to 9 mm has 50% nodal metastasis, 11% local recurrence, and 77% 5-year actuarial disease free survival; tumor of more than 9 mm has 65% nodal metastasis, 26% local recurrence, and 60% 5-year actuarial disease-free survival. CONCLUSIONS: Tumor thickness should be considered in the management of patients with oral tongue carcinoma. PMID- 11044532 TI - Extracorporeal membrane oxygenation: a ten-year experience. AB - BACKGROUND: Extracorporeal membrane oxygenation (ECMO) is a supportive therapy used for severe acute respiratory distress syndrome (ARDS). We present outcome, clinical parameters, and complications in a cohort of 245 ARDS patients of whom 62 were treated with ECMO. METHODS: Data of all ARDS patients were prospectively collected between 1991 and 1999. Outcome and clinical parameters of patients treated with and without ECMO were evaluated. RESULTS: One hundred thirty-eight patients were referred from other hospitals, 107 were primarily located in our hospital. About one fourth of these patients were treated with ECMO. The survival rate was 55% in ECMO patients and 61% in non-ECMO patients. CONCLUSIONS: ECMO is a therapeutic option for patients with severe ARDS, likely to increase survival. However, a randomized controlled study proving its benefit is still awaited. Until the development of a causal or otherwise superior therapy ECMO should be used in selected patients. PMID- 11044533 TI - Paraganglioma of the carotid bifurcation. PMID- 11044534 TI - Estimation of cord blood erythropoietin in pre-eclampsia and eclampsia. AB - OBJECTIVE: The estimation of cord blood erythropoietin in subjects with pre eclampsia and eclampsia. METHOD: Erythropoietin was measured, using ELISA, in the cord blood of infants born to 83 mothers with pre-eclampsia, and 7 with eclampsia. Another 90 subjects with no evidence of pre-eclampsia or eclampsia were taken as control subjects. Maternal parity, gestational age, blood pressures, 24-h urine protein and Apgar scores of the infants delivered were also noted. RESULT: Cord blood erythropoietin levels were statistically significantly higher (P<0.001) in infants born to mothers with pre-eclampsia and eclampsia. There was a significant positive correlation (P<0.01) between cord blood erythropoietin levels and maternal blood pressure (systolic and diastolic) and albuminuria. A negative correlation (P<0.01) was observed with the birth weights of infants. CONCLUSION: Pre-eclampsia and eclampsia are associated with higher levels of cord blood erythropoietin. PMID- 11044535 TI - High concentrations of serum inhibin in pre-eclampsia. AB - OBJECTIVE: To evaluate maternal serum immunoreactive inhibin (ir-inhibin) concentrations in women with pre-eclampsia, and assess the correlation between serum ir-inhibin and HCG. METHODS: The subjects comprised 28 pregnant women with suspected intrauterine growth retardation (IUGR) during the third trimester. Serum concentrations of ir-inhibin and HCG were measured in 13 women with pre eclampsia and 15 pregnant women as control subjects. Serum ir-inhibin was determined by a double antibody radioimmunoassay, and HCG by a solid-phase immunoradiometric assay. RESULTS: There were no significant differences in maternal characteristics between the pre-eclamptic group and control group. The pre-eclamptic group had significantly higher concentrations of serum ir-inhibin and HCG compared with the control group. The serum concentrations of ir-inhibin correlated positively with those of HCG. CONCLUSION: The pre-eclamptic patients displayed high serum levels of ir-inhibin and HCG, and this might reflect hyperplasia of trophoblastic cells. PMID- 11044536 TI - Induction of labor by prostaglandin E(2) in relation to the Bishop score. AB - OBJECTIVE: To compare the effectiveness of prostaglandin E(2) intravaginal gel with the intracervical gel in patients with an unfavorable cervix. METHOD: In a prospective multicenter trial 470 patients with unfavorable Bishop scores (3-4) were randomized to receive prostaglandin vaginal gel (2 mg) or intracervical gel (0.5 mg). RESULTS: In patients with unfavorable Bishop scores the intravaginal application route resulted in a better cervical ripening, a shorter induction to delivery interval and a higher cumulative rate of deliveries during 24 h (P=0.01). CONCLUSION: Intravaginal instillation of prostaglandin E(2) gel for induction of labor is effective in patients with an unfavorable Bishop score of 3 4. PMID- 11044537 TI - Prophylaxis and treatment of thromboembolic diseases during pregnancy with dalteparin. AB - OBJECTIVES: To determine if a prophylactic dose of dalteparin, 5000 IU daily, and if the adjusted-weight dalteparin therapeutic dose of 100 IU/kg twice daily are appropriate in pregnancy. METHOD: Anti-Xa activity levels were used to assess prophylactic (33 women) and therapeutic (15 women) dalteparin dosage throughout pregnancy. Analysis of variance was used and P-values less than 0.05 were considered statistically significant. RESULTS: In the prophylactic group, anti-Xa activity levels did not vary significantly throughout pregnancy (P=0.15). The initial dalteparin dose was modified on the first anti-Xa activity measurement in eight women, whose weight was statistically different from those remaining on the initial dose (P<0.001). The adjusted-weight therapeutic dalteparin dose induced adequate anti-Xa activity levels. CONCLUSIONS: Dalteparin, 5000 IU daily, is suitable for most pregnant women and does not need to be modified in the third trimester. A therapeutic dalteparin dose adjusted according to pregnancy weight is appropriate. PMID- 11044538 TI - Oral contraceptives do not affect magnesium in breast milk. AB - OBJECTIVES: To study the effects of oral contraceptives on magnesium metabolism of lactating mothers. METHODS: Serum and breast-milk magnesium were measured in three groups of mothers that were using combination pill (12), mini-pill (21), and a control group (21). RESULTS: There was no significant effect of oral contraceptives on breast-milk magnesium. CONCLUSIONS: The use of oral contraceptives such as the combination pill and mini-pill does not seem to affect the secretion of magnesium in breast milk. PMID- 11044539 TI - Six thousand office diagnostic-operative hysteroscopies. AB - OBJECTIVE: To compare CO(2) and normal saline as distention media in office diagnostic hysteroscopy. METHODS: The outcome of more than 6000 office hysteroscopies was analyzed. We used carbon dioxide or saline as distension medium. Minor hysteroscopic techniques were performed when indicated. RESULTS: The major indication was abnormal uterine bleeding (45%). Satisfactory hysteroscopy was achieved in 92. 4% with CO(2) and in 98.3% with saline (P<0.05). Local anesthesia was used in 54 patients (1.5%) with CO(2) and in three patients (0. 1%) with saline (P<0.001). Four hundred and two women (16.3%) underwent hysteroscopic procedures under saline hysteroscopy. Endometrial polyps were removed in 281 patients, 75 IUDs were removed, 14 fibroids were extracted, uterine septa were excised in 11 cases and mild and moderate adhesions were transected in 21 patients. CONCLUSION: Saline office diagnostic hysteroscopy offers at least all the advantages of the CO(2) hysteroscopy, and gives the possibility to easily 'find and treat in situ' many of the lesions observed. PMID- 11044540 TI - beta(2)-adrenergic agonists and pelvic floor exercises for female stress incontinence. AB - OBJECTIVE: We compared beta(2)-adrenergic agonist therapy with clenbuterol (DT) and physiological therapy (PT) in a randomized study to establish the first line therapy for stress incontinence (SI). METHOD: The clinical efficacy of DT (group A), PT (group B), and a combination of DT and PT (group C) was investigated in 61 patients with SI by means of a 12-week randomized controlled study. The frequency and volume of SI and the patients' own impressions were used as the basis for the assessment of efficacy. RESULTS: The SI improvement rates in groups A, B, and C were 76.9, 52.6, and 89. 5%, respectively (P=0.0361). A significant therapeutic effect on the frequency of SI was observed in group B and group C at 2 weeks after the start of treatment (both P<0.05), and in all groups at 6 weeks (all P<0.01). The efficacy rates based on the patients' own impressions in groups A, B, and C were 84.6, 31.6, and 68.4%, respectively (P=0.0064). CONCLUSION: The beta(2)-adrenergic agonist appeared to be clinically useful as a drug of choice for SI. PMID- 11044541 TI - High risk HPV and p53 protein expression in cervical intraepithelial neoplasia. AB - INTRODUCTION: Cervical intraepithelial lesions due to HPV infection are common in Brazil. An understanding of the mechanisms of the interaction between HPV and host factors is still incomplete. In spite of the high incidence of cervical cancer in Brazil, such studies with Brazilian patients are scarce. The purpose of this study was to correlate the presence of high-risk types of HPV and expression of p53 protein, grade of cervical lesion, age, high-risk sexual behaviors and smoking. It was also intended to establish whether p53 expression might be useful as a marker for CIN progression. METHODS: HPV detection was performed on paraffin sections using biotin-labeled probes by in situ hybridization. p53 protein expression was evaluated by immunohistochemistry. RESULTS: Seventy-eight patients with cervical dysplasia were included in the study. CIN 1 was diagnosed in 38 cases, and CIN 2+3 in 40 cases. High-risk HPV was detected in 42 patients. No correlation was found between the grade of cervical lesion or the presence of HPV and smoking, and high-risk sexual behavior. Expression of p53 was significantly higher in CIN 1, as compared with CIN 2+3, but did not correlate with HPV status. CONCLUSION: Higher expression of p53 protein in early lesions supports the hypothesis of a partially protective role of the wild-type p53 in early stages of cervical lesions. PMID- 11044542 TI - Effects of estrogen and alendronate on sister chromatid exchange (SCE) frequencies in postmenopausal osteoporosis patients. AB - OBJECTIVE: To compare two types of osteoporosis treatment for genotoxicity by using sister chromatid exchange (SCE) frequencies. METHOD: Fifty-seven women, aged between 40 and 64 years, composed the population in the study. SCE values of patients under estrogen replacement therapy (ERT) or alendronate therapy were compared to controls who never used any drugs for osteoporosis. RESULT: The difference between the SCE values of women taking ERT and control women was found to be statistically significant (P<0.05). The difference between women taking alendronate and untreated controls was not statistically significant. CONCLUSION: Our results indicate that alendronate does not have genotoxic effects based on SCE frequency, while ERT increases SCE frequencies. PMID- 11044543 TI - Vaginal misoprostol in the management of first-trimester missed abortions. AB - OBJECTIVE: To evaluate the efficacy of a regimen of vaginal misoprostol in causing the complete expulsion of first-trimester missed abortions, or alternatively dilating the cervix for surgical evacuation. METHOD: Seventy-four women with a transvaginal ultrasound diagnosis of a first-trimester missed abortion and no more than slight vaginal bleeding were consecutively enrolled. Misoprostol (600 microg) was administered vaginally and repeated 4 h later if necessary. Surgical evacuation was performed when complete expulsion was not documented on the ultrasound 10-12 h after treatment. RESULTS: Complete medical evacuation occurred in 42 women (56.8%), 11 (14.9%) of which required only one dose. Seventy women (94.6%) experienced abdominal pain, 73 (98.6%) vaginal bleeding, 10 (13.5%) nausea, 4 (5.4%) vomiting, 5 (6.8%) diarrhea, and 4 (5.4%) transient hyperthermia. There was one case of heavy vaginal bleeding requiring emergency surgical evacuation, and one re-admission for incomplete abortion at 30 days. All but 4 (5.4%) women had permeable cervices at the time of surgery. CONCLUSION: The described regimen of vaginal misoprostol is safe and reasonably effective in inducing complete evacuation in missed abortions. When this does not occur, it almost always provides adequate cervical dilatation for surgery. PMID- 11044544 TI - Twin pregnancy following gonadotrophin therapy in a patient with Sheehan's syndrome. AB - A case of Sheehan's syndrome presented with secondary amenorrhea and was put on L thyroxine, prednisolone and cyclical estrogen and progestin. Ovulation induction with gonadotrophins and intrauterine insemination with husband's semen resulted in a twin pregnancy. Antepartum course was complicated by bronchial asthma, gestational diabetes and pregnancy-induced hypertension. Cesarian section was done at 34 weeks gestation for preterm rupture of membranes and breech presentation. Both babies and their mother were doing well at 6 months of follow up. PMID- 11044545 TI - Lumbar lordosis and the spread of subarachnoid hyperbaric 0.5% bupivacaine at cesarean section. PMID- 11044546 TI - Antenatal detection of intrauterine growth restriction in patients with systemic lupus erythematosus. PMID- 11044547 TI - Accuracy of the blood loss estimation in the third stage of labor. PMID- 11044548 TI - The scope and limits of conscientious objection. AB - Principles of religious freedom protect physicians, nurses and others who refuse participation in medical procedures to which they hold conscientious objections. However, they cannot decline participation in procedures to save life or continuing health. Physicians who refuse to perform procedures on religious grounds must refer their patients to non-objecting practitioners. When physicians refuse to accept applicants as patients for procedures to which they object, governmental healthcare administrators must ensure that non-objecting providers are reasonably accessible. Nurses' conscientious objections to participate directly in procedures they find religiously offensive should be accommodated, but nurses cannot object to giving patients indirect aid. Medical and nursing students cannot object to be educated about procedures in which they would not participate, but may object to having to perform them under supervision. Hospitals cannot usually claim an institutional conscientious objection, nor discriminate against potential staff applicants who would not object to participation in particular procedures. PMID- 11044549 TI - This is a selection of abstracts taken from the literature in the field of gynecology and obstetrics which the Journal' s editors feel may be of interest to our readers(1) PMID- 11044551 TI - A comparison of DNA vaccines expressing the 45W, 18k and 16k host-protective antigens of Taenia ovis in mice and sheep. AB - The immunogenicity of DNA vaccines encoding three different Taenia ovis host protective antigens was compared in mice and sheep. DNA vaccines encoding the 45W, 18k and 16k antigens of T. ovis were constructed. The ability of DNA vaccines encoding the 45W and 18k genes to express antigen was confirmed by Western blotting of transfected Cos-7 cells. BALB/c mice were vaccinated intramuscularly with 45W, 18k or 16k DNA vaccines and the humoral immune response analysed by ELISA. DNA vaccines expressing 45W, 18k or 16k antigen were immunogenic in mice and generated significant titres of antigen-specific antibody. Intramuscular vaccination of outbred sheep with the T. ovis DNA vaccines generated significantly lower titres of 45W-specific antibody and failed to generate 18k or 16k-specific antibody. The findings of this study show that each of the three T. ovis host-protective antigens are amenable to delivery via DNA vaccines, and that the parameters governing the efficacy of DNA vaccines in sheep require further investigation. PMID- 11044550 TI - Atherosclerosis: from lesion formation to plaque activation and endothelial dysfunction. AB - Atherosclerosis is an important source of morbidity and mortality in the developed world. Despite the fact that the association between LDL cholesterol and atherosclerosis has been evident for at least three decades, our understanding of exactly how LDL precipitates atherosclerosis is still in its infancy. At least three working hypotheses of atherosclerosis are now nearing the stage where their critical evaluation is possible through a combination of basic science investigation and murine models of atherosclerosis. As we move forward in our understanding of this disease, efforts will be increasingly focused on the molecular mechanisms of disease activation that precipitate the clinical manifestations of atherosclerosis such as heart attack and stroke. Two candidates for such investigation involve the events surrounding plaque activation and endothelial dysfunction. Further investigation in these fields should provide the necessary insight to develop the next generation of interventions that will reduce the clinical manifestations of this devastating disease. The purpose of this work is to review the major theories of atherogenesis, examine the aspects of atherosclerosis that lead to disease activation and discuss aspects of disease activation that are amenable to treatment. PMID- 11044552 TI - Detection of cytokines in bovine colostrum. AB - Colostrum contains factors that are protective for the neonate and may be a source of immunomodulary molecules that positively influence the immune status of the neonate. To confirm that colostrum contains a variety of cytokines with immunomodulatory properties, we established a bovine cytokine specific ELISA and five cytokines (IL-1 beta, IL-6, TNF-alpha, INF-gamma or IL-1 receptor antagonist, IL-1ra) in the whey samples from cows at different stages of lactation were monitored. The expression of cytokine mRNAs (IL-1 beta, IL-6, TNF alpha and INF-gamma) in the colostral cells was detected by RT-PCR. The concentrations of cytokines in colostrum were significantly higher concentrations than those in the mature milk. A positive correlation was observed between the concentrations of IL-1ra and IL-1 beta in the colostrum samples. In conclusion, colostrum contains high levels of cytokines that could be produced and secreted in the mammary gland and that may have an immunomodulatory activity and influence neonatal immunity. PMID- 11044553 TI - In vitro effects of beta-hydroxy-beta-methylbutyrate (HMB) on cell-mediated immunity in fish. AB - beta-Hydroxy-beta-methyl butyrate(HMB) has been shown to counteract many of the negative effects of intensive animal production methods and results in increased growth and protection against diseases. In the present study, the effect of HMB on the immunocompetence cell activity in rainbow trout (Oncorhynchus mykiss) and carp (Cyprinus carpio) was examined. Pronephric phagocytes and lymphocytes were isolated from the fish and grown in culture medium (RPMI-1640) containing either 0, 0.1, 1, 5, 10, 25, 50 or 100 microg HMB/ml of medium. The effects of HMB on the respiratory burst activity (RBA) stimulated by phorbol myristate acetate (PMA), the potential killing activity (PKA) and lymphocyte proliferation stimulated by either concanavalin A (Con-A) or lipopolysaccharide (LPS) were examined. The addition of HMB to the culture medium increased the RBA by up to 84% (p<0.01) over that of cells grown without HMB. Similarly, the PKA of the phagocytes was also increased with HMB addition to the medium by up to 140% (p<0.01) over that of cells grown without HMB. Lymphocyte proliferation stimulated by both ConA and LPS was also increased approximately two-fold (p<0.01) when HMB was added to the culture medium at concentrations between 10 and 100 microg HMB/ml in both rainbow trout and carp. The greatest effects of HMB on RBA and PKA activities were observed at a concentration >50 microg HMB/ml while lymphocyte proliferation was maximally stimulated at 25 microg HMB/ml. In conclusion, the current study shows that HMB could potentially improve immunocompetence cell activity in fish through increased cell proliferation and functionality. PMID- 11044554 TI - Increased pulmonary secretion of tumor necrosis factor-alpha in calves experimentally infected with bovine respiratory syncytial virus. AB - Bovine respiratory syncytial virus (BRSV) is an important cause of respiratory disease among calves in the Danish cattle industry. An experimental BRSV infection model was used to study the pathogenesis of the disease in calves. Broncho alveolar lung lavage (BAL) was performed on 28 Jersey calves, of which 23 were experimentally infected with BRSV and five were given a mock inoculum. The presence of the cytokine tumor necrosis factor alpha (TNF-alpha) in the BAL fluids was detected and quantified by a capture ELISA. TNF-alpha was detected in 21 of the infected animals. The amount of TNF-alpha in the BAL fluid of calves killed post inoculation day (PID) 2 and 4 was at the same very low level as in the uninfected control animals. Large amounts of TNF-alpha were detected on PID 6, maximum levels of TNF-alpha were reached on PID 7, and smaller amounts of TNF alpha were seen on PID 8. The high levels of TNF-alpha appeared on the days where severe lung lesions and clinical signs were obvious and the amounts of BRSV antigen were at their greatest. Although Pasteurellaceae were isolated from some of the BRSV-infected calves, calves treated with antibiotics before and through the whole period of the infection, as well as BRSV-infected calves free of bacteria reached the same level of TNF-alpha as animals from which bacteria were isolated from the lungs. It is concluded that significant quantities of TNF-alpha are produced in the lungs of the calves on PID 6-7 of BRSV infection. The involvement of TNF-alpha in the pathogenesis of, as well as the anti-viral immune response against, BRSV infection is discussed. PMID- 11044555 TI - MHC class II-restricted, CD4(+) T-cell proliferative responses of peripheral blood mononuclear cells from Mycobacterium bovis-infected white-tailed deer. AB - White-tailed deer are significant wildlife reservoirs of Mycobacterium bovis for cattle, predators, and, potentially, humans. Infection of cattle with M. bovis stimulates an antigen-specific T-cell response, with both CD4(+) and CD8(+) cells implicated in protective immunity. Few studies, however, have examined lymphocyte subset responses to experimental M. bovis infection of white-tailed deer. In this study, a flow cytometric proliferation assay was used to determine the relative contribution of individual peripheral blood mononuclear cell subsets of M. bovis infected white-tailed deer in the recall response to M. bovis antigen. Naive deer were challenged with M. bovis by cohabitation with infected deer. These M. bovis challenged deer developed significant in vivo (delayed-type hypersensitivity) and in vitro (proliferative) responses to M. bovis purified protein derivative (PPD). At necropsy, typical tuberculous lesions containing M. bovis were detected within lungs and lung-associated lymph nodes of infected deer. The predominant subset of lymphocytes that proliferated in response to in vitro stimulation with PPD was the CD4(+) subset. Minimal proliferative responses were detected from CD8(+), gamma delta TCR(+), and B-cells. Addition of monoclonal antibodies specific for MHC II antigens, but not MHC I or CD1 antigens, abrogated the proliferative response. Together, these findings indicate that while CD4(+) cells from infected deer proliferate in the recall response to M. bovis antigens, this response is not sufficient to clear M. bovis and immunologic intervention may require stimulation of alternate subsets of lymphocytes. PMID- 11044556 TI - Presence of glutamine at position 74 of pocket 4 in the BoLA-DR antigen binding groove is associated with occurrence of clinical mastitis caused by Staphylococcus species. AB - Potential relationships between amino acid motifs in the antigen binding groove of various alleles of the bovine major histocompatibility complex DR (BoLA-DR) molecule and occurrence of clinical mastitis caused by Staphylococcus species (non-Staphylococcus aureus) were investigated in a case-control study. A significant association (P< or =0.05) was detected between the presence of glutamic acid at position beta 74 and occurrence of mastitis caused by Staphylococcus spp. with a relative risk of 11. This motif is present in BoLA DRB3.2*22, *23 and *24 alleles. Presence of a positively charged residue (arginine or lysine) at position 13 also showed a tendency (P< o r=0.1) towards an association with a higher risk of clinical mastitis caused by the same bacteria. This motif is present in BoLA-DRB3.2*23 and *8 alleles. Similarly, presence of arginine at position beta 71 (present in alleles *23 and *22) was associated with occurrence of this disease. These positions (beta 13, beta 71 and beta 74) form pocket 4 of the antigen binding groove, which plays an instrumental role in antigen binding and recognition by T-lymphocytes. Thus, it can be concluded that pocket 4 of the BoLA-DR molecule is involved in conferring susceptibility to clinical mastitis caused by Staphylococcus spp. PMID- 11044557 TI - Effect of an intravitreal cyclosporine implant on experimental uveitis in horses. AB - The purpose of this study was to determine the effects of an intravitreal device releasing cyclosporine A (CsA) on recurrent inflammatory episodes in experimental uveitis. Nine normal horses were immunized peripherally with H37RA-mTB antigen twice, and then received 25 microg of H37RA-mTB antigen intravitreally in the right eye and an equal volume of balanced salt solution intravitreally in the left eye. Two weeks later, the animals randomly received either a CsA or a polymer implant (without CsA) in both eyes 1 week following implantation of the devices, 25 microg of H37RA-mTB antigen was reinjected into the right eye of each animal. Clinical signs of ophthalmic inflammation were graded following injections and implantation. The animals from each group were euthanized at 3, 14, and 28 days following the second injection. Aqueous and vitreous humor protein concentrations were measured. The presence, number, and type (CD4, 5 and 8) of infiltrating inflammatory cells and amount of tissue destruction were determined. Total RNA was isolated and quantitative reverse transcriptase polymerase chain reaction was performed for equine specific interleukin (IL) 2 and 4, interferon-gamma (IFN gamma) and beta-actin. In addition, aqueous and vitreous humor and peripheral blood were collected at the termination of the experiments and analyzed for CsA concentration by HPLC. Within 4h of the first intravitreal H37RA-mTB antigen injection, each animal developed epiphora, blepharospasm, mild corneal edema, aqueous flare, myosis, and vitreous opacity. The severity of signs peaked 48 to 72 h after injection and subsequently decreased back to normal within 14 days. Following the second injection, clinical signs in the eyes with the CsA device were less severe and significantly shorter in duration than signs with the polymer only implant eyes. Aqueous and vitreous humor protein levels, infiltrating cell numbers, total number of T-lymphocytes, and levels of IL-2 and IFN gamma-mRNA were significantly less in eyes with the CsA implant compared to eyes with the polymer only. CsA implants did not completely eliminate the development of a second ('recurrent') experimental inflammatory episode in these horses. However, the duration and severity of inflammation, cellular infiltration, tissue destruction, and pro-inflammatory cytokines RNA transcript levels were significantly less in those eyes implanted with the CsA device. PMID- 11044558 TI - The immunogenicity and efficacy of replication-defective and replication competent bovine adenovirus-3 expressing bovine herpesvirus-1 glycoprotein gD in cattle. AB - Replication-competent and replication-defective bovine adenovirus type 3 recombinants expressing the bovine herpesvirus type 1 (BHV-1) glycoprotein D (gD) were tested for induction of gD specific immune responses in calves using intratracheal (1st and 2nd immunization) and sub-cutaneous (3rd immunization) route of immunization. The replication-defective recombinant BAV501 induced systemic immune responses against gD as low titers of anti gD-IgG were detected in the serum. However, the efficacy of the replication-competent BAV3.E3gD to induce gD-specific antibodies in the serum and the nasal secretions was superior to that of replication-defective BAV501 when both viruses were given at the same dosage. Partial protection from challenge was induced in calves immunized with replication-competent BAV3.E3gD. A dramatic increase in the titers of anti-gD IgG and IgA levels, both in serum and nasal secretions, following BHV-1 challenge (anamnestic response) suggested that the animals immunized with replication defective BAV501 had been primed for gD-specific antibody responses. PMID- 11044559 TI - Immune and clinical parameters associated with Leishmania infantum infection in the golden hamster model. AB - For experimental infections with viscerotropic strains of Leishmania, a suitable animal model is not yet defined. In the present work, we have reappraised the use of golden hamster (Mesocricetus auratus) as an experimental model for infection with Leishmania infantum. Groups of hamsters were challenged by the intracardial route with doses ranging from 10(3) to 10(5) infectious promastigotes and the animals were monitored for 1-year follow-up period. The outcome of the infection was assessed by clinical symptoms of leishmaniasis, parasite loads in both liver and spleen, humoral response to Leishmania antigens and antibody levels in kidneys. The humoral response was analysed using either crude antigens (by ELISA and Western blotting) or several recombinant Leishmania antigens (Hsp70, Hsp83, LiP2a, LiP2b, H2A, H3 and KMP-11). From the analysis of all these parameters, we established the existence of three groups of animals: symptomatic or susceptible, oligosymptomatic, and resistant. Given the parallelism existing between the outcomes of Leishmania-infection in hamsters, dogs and humans, we believe that our data illustrate that the hamster is an excellent experimental model to study visceral leishmaniasis and for the design of vaccine development. PMID- 11044560 TI - Cloning of equine chemokines eotaxin, monocyte chemoattractant protein (MCP)-1, MCP-2 and MCP-4, mRNA expression in tissues and induction by IL-4 in dermal fibroblasts. AB - We report the cloning of four equine CC chemokines, eotaxin, monocyte chemoattractant protein (MCP)-1, MCP-2 and MCP-4, which show high levels of identity with their respective homologous sequences in other species. Using a multiplex RT-PCR, we have studied the constitutive mRNA expression of these four CC chemokines in skin, lung, liver, spleen, jejunum, colon and kidney of normal adult horses and compared this data with the eosinophil counts in the same samples. We demonstrate that eotaxin mRNA is only expressed in jejunum and colon, where there are large numbers of eosinophils suggesting that eotaxin might be recruiting eosinophils in the normal digestive tract of the horse. MCP-1 and MCP 4 are expressed in all tissues whereas MCP-2 is only found in some samples of lung, spleen, liver and kidney. We also report the early induction (2h) of equine eotaxin and MCP-4, and the up-regulation of MCP-1 by interleukin-4 in dermal fibroblasts, suggesting these chemokines might be involved in equine skin allergic diseases. PMID- 11044561 TI - Molecular cloning and phylogenetic analysis of a cDNA encoding the cat (Felis domesticus) Ig epsilon constant region. AB - A feline splenic cDNA library was screened with a (32)P-labelled cDNA probe encoding the canine IgE epsilon heavy chain subunit. A cDNA sequence of 1614 nucleotides encoding the complete feline IgE heavy chain, as well as a portion of a variable region, was identified. A search of the GenBank database revealed an identity of 82% at the nucleotide level and 76% at the amino acid level between the feline epsilon heavy chain sequence and the canine homologue. In a separate study, feline genomic DNA, isolated from whole feline embryo cells, was subjected to PCR amplification using primers based on known partial genomic DNA sequences for the feline C epsilon gene. Following removal of an intron from the 683 bp PCR product, the coding sequence yielded an ORF of 506 bp. The DNA sequence of this PCR clone differed by a single nucleotide from the cDNA clone. This difference is silent, and therefore the proteins encoded by the two sequences are identical over the regions cloned and sequenced. Phylogenetic analysis of the constant regions of nine immunoglobulin epsilon genes revealed that the feline cDNA is most similar to the canine homologue. PMID- 11044562 TI - Excretion patterns of mucosally delivered antibodies to p23 in Cryptosporidium parvum infected calves. AB - Fecal samples obtained at intervals from six calves with acute cryptosporidiosis contained antibodies of multiple isotypes to p23. IgM-, IgA-, and IgG(1)-isotype anti-p23 appeared before IgG(2)-isotype antibodies. All anti-p23 antibodies had declined by 2 months after infection. One calf that failed to shed oocysts following initial exposure developed IgG(1)-isotype anti-p23 antibodies. One calf that died following exposure to Cryptosporidium parvum oocysts lacked detectable anti-p23 antibodies. Re-inoculation with C. parvum resulted in a brief, marked recall response to p23. PMID- 11044563 TI - Phosphodiesterase activity in neutrophils from horses with chronic obstructive pulmonary disease. AB - Neutrophils are recruited to the lungs of horses with chronic obstructive pulmonary disease (COPD) and exhibit increased activity after antigen challenge. Phosphodiesterase type4 (PDE4) inhibitors have been shown to attenuate human neutrophil activation. The aim of this study was to establish the PDE isoenzyme profile of equine neutrophils using isoenzyme selective inhibitors to determine if these compounds should be evaluated in horses with COPD. Total cAMP and cGMP dependent PDE activity was no different in neutrophils from normal (156.2+/-7.1 and 6.8+/-0.6 pmol/min/mg for cAMP and cGMP, respectively) and COPD susceptible horses (146.0+/-10.2 and 5.5+/-0.6 pmol/min/mg for cAMP and cGMP, respectively). The PDE4 inhibitors, CDP840 and rolipram, caused significant, concentration related and almost complete inhibition of PDE activity (IC(50) values=8.8+/-0.1 x 10(-9) and 7.3+/-0.2 x 10(-9)M for CDP840; 1.2+/-0.1 x 10(-6) and 1.1+/-0.1 x 10( 6)M for rolipram in normal and COPD susceptible horses, respectively). The inhibitory effects of the mixed PDE3/ PDE4 inhibitor, zardaverine were of similar magnitude and potency to rolipram. However, the limited inhibitory effects of the PDE3 inhibitor, siguazodan, suggest that zardaverine is acting primarily via PDE4 inhibition. These results indicate that PDE4 is the predominant isoenzyme present in the equine neutrophil and inhibition of PDE activity using selective PDE4 inhibitors may, therefore, modulate equine neutrophil activation in horses with COPD. PMID- 11044564 TI - A new tannase substrate for spectrophotometric assay. AB - A new tannase substrate, protocatechuic acid p-nitrophenyl ester, 5, was synthesized using modern synthetic methods. The synthesis was designed to be performed by non-specialized chemists. It only involves four steps, three of which are protection-deprotection, and uses standard methods of separation and purification, such as recrystallization and column chromatography over silica. Under tannase action, protocatechuic acid p-nitrophenyl ester, 5, releases p nitrophenol, which is easily measured spectrophotometrically either at 350 nm for pH values<6 or at 400 nm for pH values of 6-7 (yellow). The pH-response and the catalytic parameters of a crude Penicillium sp. tannase preparation were determined using 5 as substrate, thus showing the usefulness of this substrate in determining tannase activity. PMID- 11044565 TI - A simple rotating annular reactor for replicated biofilm studies. AB - The performance of two types of rotating annular reactors for the cultivation of river biofilms was compared qualitatively and quantitatively. One reactor was a commercially available system with a rotating inner solid cylinder and polycarbonate slides in the outer fixed cylinder. The other, a non-commercial system manufactured in the laboratory, had the polycarbonate slides positioned on a machined, rotating inner cylinder. Microscale comparison of the biofilms was carried out using confocal laser scanning microscopy techniques including, fluorescent nucleic acid staining, fluor conjugated lectins and autofluorescence imaging. The results obtained indicated that the reactors were similar in terms of biofilm development pattern, thickness, bacterial biomass, and exopolymer production. Significant differences were found in terms of photosynthetic biomass with the glass bodied non-commercial reactor providing more favourable conditions for algal growth than the opaque polycarbonate outer cylinder of the commercial reactor. The study indicated that a simple inexpensive reactor constructed from available components and materials, produced river biofilms similar to those obtained using a commercial system but at substantially lower cost. The availability of such inexpensive annular reactors should facilitate much needed replicated studies of biofilm development. PMID- 11044566 TI - A comparative study on the disintegration of filamentous fungi. AB - Different methods for cell disintegration were tested for their efficacy on filamentous fungi, including percussion grinding, homogenization using an Ultra Turrax, chemical treatment and lyophylization. The release of protein from Ganoderma applanatum and Pycnoporus cinnabarinus and the activity of cytoplasmatic glucose-6-phosphate dehydrogenase in the crude extracts were monitored to determine the efficiency of each disintegration technique used. Fungal cells proved to be particularly resistant towards some disintegration methods commonly used for yeasts and bacteria. Best results were obtained using a percussion grinder, if necessary, in combination with an Ultra-Turrax pretreatment. PMID- 11044568 TI - Rapid detection, identification, and enumeration of Pseudomonas aeruginosa in bottled water using peptide nucleic acid probes. AB - A new chemiluminescent in situ hybridization (CISH) method that provides simultaneous detection, identification, and enumeration of Pseudomonas aeruginosa in bottled water within 1 working day has been developed. Individual micro colonies of P. aeruginosa were detected directly on membrane filters following 5 h of growth by use of soybean peroxidase-labeled peptide nucleic acid (PNA) probes targeted to a species-specific sequence in P. aeruginosa rRNA. Within each micro-colony, reaction of the peroxidase with a chemiluminescent substrate generated light that was subsequently captured by film or with a digital camera system. Each spot of light represented one micro-colony of P. aeruginosa. Sensitivity and specificity for the identification of P. aeruginosa were 100% as determined by testing 28 P. aeruginosa strains and 17 other bacterial species that included closely related Pseudomonas species. Furthermore, the number of micro-colonies of P. aeruginosa represented by light spots correlated with counts of visible colonies following sustained growth. We conclude that PNA CISH speeds up traditional membrane filtration techniques and adds the specificity of PNA probe technology to generate fast and definitive results. PMID- 11044567 TI - Rapid detection of fluorescent and chemiluminescent total coliforms and Escherichia coli on membrane filters. AB - The detection of fluorescent colonies of Escherichia coli/total coliforms (TC) on a membrane filter is currently carried out using 4-methylumbelliferyl-beta-D glycosides as enzyme substrates and a UV-lamp for visualization. The most rapid procedures based on this approach for the demonstration of these indicator bacteria in water take 6-7.5 h to complete. As part of efforts to further reduce the detection time, an improved two-step procedure for the fluorescence or chemiluminescence labelling of microcolonies of E. coli/TC on a membrane filter has been developed. Essential features of this approach include a separation of the bacterial propagation and target enzyme induction from the actual enzymatic labelling, the use of improved fluorogenic, i.e., 4-trifluoromethylumbelliferyl beta-D-glycosides and fluorescein-di-beta-D-glycosides, or chemiluminogenic (i.e., phenylglucuronic- or galactose-substituted adamantyl 1,2-dioxetanes) substrates for beta-glucuronidase/beta-galactosidase, of enzyme inducers, of special membrane filters and of polymyxin B to promote the cellular uptake of the substrate. This labelling procedure has been applied in conjunction with different detection devices including a UV-lamp, CCD-cameras, X-ray film and the ChemScan((R)) RDI. Using the former three, microcolonies of pure cultures could be detected within 5.5-6.5 h, but waterborne E. coli/TC may fail to form microcolonies in this short time period, thus yielding poor sensitivity and a high false-negative rate. In contrast, a quantitative enumeration was feasible in less than 4 h with the ChemScan((R)) RDI, owing to its ability to detect both microcolonies and non-dividing single cells. PMID- 11044569 TI - Flow cytometry characterisation of Salmonella typhimurium mutants defective in proton translocating proteins and stationary-phase growth phenotype. AB - We have shown that the growth, starvation and population heterogeneity of Salmonella typhimurium and its isogenic nuoG and cydA mutants can be monitored by flow cytometry. Bacterial cells were analysed unstained, and after staining with rhodamine 123, propidium iodide and acridine orange. In unstained cultures it was possible to distinguish flagellated and non-flagellated cells. nuoG and cydA mutants were less stained with rhodamine confirming their defects in generating membrane potential. Increase in propidium iodide staining associated with reduced membrane integrity was seen between day 4 and 14 in all the strains. Acridine orange staining showed that there was retarded development in stationary phase in nuoG and cydA mutants. Furthermore, up to day 28, a small portion of cells showed high RNA and DNA levels. To determine whether these cells represent a sub population better adapted for long term survival, we measured the growth of the population by both OD values and viable counts. Because the OD values increased throughout the whole study in both wild-type and mutant strains, while the viable counts gradually decreased, we propose that even in very old cultures there must be a population of cells undergoing replication. PMID- 11044570 TI - The use of serologic tests for the diagnosis of chlamydial infections. AB - Serology is commonly used for the diagnosis of acute Chlamydia pneumoniae infections and also for the diagnosis of complicated Chlamydia trachomatis infections. Furthermore, recent sero-epidemiological studies have linked C. pneumoniae infection with several diseases traditionally considered non infectious. The objectives of this mini-review are to critically review and discuss some selected analytical and methodological aspects, controversies and current problems in chlamydial serodiagnosis. To illustrate our views we present some original data of the comparison of current technologies. The review of the literature revealed high variability in methodologies applied to different studies. This observation was supported by our own data, which explains occasional conflicting clinical interpretation. Although the microimmunofluorescence (MIF) technique is generally considered as the gold standard for serodiagnosis of chlamydial infections, assay conditions are highly variable and hence pose a major problem in the interpretation of the results. For instance, many recent studies linking C. pneumoniae and atherosclerosis have utilized MIF techniques with variable threshold criteria for the positivity, in combination with selection bias of cases and controls possibly leading to conflicting results. Variability of assay conditions is also a common problem with Western blots, and interpretation is problematic when both anti-C. pneumoniae and anti-C. trachomatis antibodies are present. Furthermore, there is a lot of disagreement in serological criteria applied to recently emerged enzyme immunoassay (EIA) techniques when these assays are used for acute and non-acute clinical conditions and their association with Chlamydiae. In conclusion, standardization of serological techniques and the development of uniform criteria for interpretation of serologic findings is necessary to increase our knowledge of the biology of Chlamydiae, pathogenesis of any chlamydial infection and chronic infections in particular. PMID- 11044571 TI - Method to sensitize bacterial spores to subsequent killing by dry heat or ultraviolet irradiation. AB - Hydrogen peroxide and ultraviolet irradiation are known to interact synergistically for killing of bacterial spores. Synergy could be demonstrated with spores of Bacillus megaterium ATCC19213 adsorbed to filter paper strips or glass coverslips treated first with the peroxide and then dried for as long as 48 h prior to UV irradiation. This delayed action was considered to be due to absorption of the peroxide by the spores in an active but not readily vaporized form, which could become sporicidal also if the spores were heated to 50 degrees C. B. megaterium spores mixed with 0.1% (32.6 mM) H(2)O(2) solution appeared to absorb as much as 15 micromol/mg dry weight or about 0.5 mg/mg, but only a third to half of the peroxide could be recovered by water washing. A part of the unrecovered peroxide was degraded in reactions resulting in measurable production of oxygen. Degradation was not reduced by heating the spores to 65 degrees C or by azide and so appeared to be non-enzymatic. Spores of the anaerobe Clostridium sporogenes were also sensitized to ultraviolet killing by H(2)O(2) treatment followed by drying. They appear to absorb less peroxide, only about 2 micromol/mg, but had lower capacities to degrade H(2)O(2) so that nearly all of the peroxide could be recovered by washing with water. The findings presented should be helpful in the design of new methods for synergistic killing of spores by H(2)O(2) and UV irradiation or dry heat, especially involving, for example, packaging materials. PMID- 11044573 TI - Effect of magnesium on calcium-dependent brain function that prolongs ethanol induced sleeping time in mice. AB - The effect of intracerebroventricular (i.c.v.) administration of magnesium on calcium- and dopamine-dependent brain function was investigated behaviorally and biochemically. The duration of ethanol-induced sleeping time in mice was prolonged following i.c.v. administration of calcium chloride (10 micromol/kg) or dopamine (30nmol/mouse); however, it was not affected by magnesium chloride (10 or 40 micromol/kg). The ability of calcium to prolong ethanol-induced sleeping time was inhibited by the administration of magnesium chloride. The brain dopamine level in mice was significantly increased following i.c.v. administration of calcium chloride. Taking into consideration these results and those from previous studies, it is suggested that calcium enhances dopamine synthesis in the brain through a calmodulin-dependent system, and the increase in dopamine level prolongs ethanol-induced sleeping time. However, magnesium inhibits dopamine release. Therefore, magnesium may inhibit calcium-dependent brain function through dopaminergic neurons, and consequently reduce the effect of calcium on ethanol activity. PMID- 11044572 TI - Protective effect of beta-carbolines and other antioxidants on lipid peroxidation due to hydrogen peroxide in rat brain homogenates. AB - Tryptoline and pinoline are two beta-carbolines isolated from the nervous system of mammals. We investigated the ability of these compounds to prevent lipid peroxidation induced by hydrogen peroxide in rat brain homogenates. We also compared their effects with other known antioxidants including melatonin, trolox and ascorbic acid. Lipid peroxidation was assessed by measuring malonaldehyde (MDA) and 4-hydroxy-alkenals (4-HDA) concentrations in the brain homogenates. Incubation with hydrogen peroxide (5 mM) increased MDA+4-HDA levels, which were totally prevented by tryptoline, pinoline, melatonin and trolox in a concentration-dependent manner. By contrast, higher MDA-4-HDA concentrations compared with control experiments were found after incubation with ascorbic acid, thus reflecting an increase of lipid peroxidation induced by this compound. Although in vivo studies are needed, the data suggest that these beta-carbolines may be potential neuroprotective agents because of their antioxidant activities. PMID- 11044574 TI - Fluorescent imaging of nitric oxide production in neuronal varicosities associated with intraparenchymal arterioles in rat hippocampal slices. AB - The fluorescent indicator 4,5-diaminofluorescein (DAF-2) has been used to investigate the production of nitric oxide in the vicinity of intraparenchymal cerebral blood vessels. Slices of rat hippocampus 300-350 microm thick, were loaded with 5 microM DAF-2 diacetate. On exposure to light of 450-490 nm wavelength, point sources of fluorescence, 1.8+/-0.2 microm in diameter (mean+/ SEM), were observed in close apposition to the outer surface of the vascular smooth muscle wall of 10/15 arterioles. In fixed slices, resectioned and processed for nicotinamide adenine dinucleotide phosphate-dependent diaphorase, stained varicose fibres were also seen in close association with the smooth muscle wall of small arterioles. These findings suggest that tonic activity in perivascular nitrergic nerve fibres lying in close proximity to intraparenchymal microvessels may be a source of dilator tone within the parenchyma. PMID- 11044575 TI - Antisense oligonucleotide suppression of Na(+)/Ca(2+) exchanger activity in primary neurons from rat brain. AB - An antisense (AS) oligodeoxynucleotide based on a conserved sequence in the three isoforms of the Na(+)/Ca(2+) exchanger (NCX) was used to decrease expression of this Ca(2+) transporter in primary neuronal cultures. Two AS oligo applications decreased NCX activity by approximately 40% within 12-24 h, and neither sense (S) or missense (MS) oligos altered NCX activity. The reduced NCX expression was confirmed by immunoblots and enzyme-linked immunosorbent assays (ELISAs). Resting [Ca(2+)](i) levels were 20% higher in AS-treated neurons and showed a slower return to baseline levels following activation of Ca(2+) influx by N-methyl-D aspartate (NMDA). These results suggest that NCX plays a significant role in maintaining neuronal Ca(2+) homeostasis and in restoring baseline Ca(2+) levels following depolarization. PMID- 11044576 TI - Connections between the trigeminal mesencephalic nucleus and the superior colliculus in the rat. AB - Retrograde tracing methods are employed here to demonstrate that neurons in the trigeminal mesencephalic nucleus (5me) project to the superior colliculus (SC) in the rat. These neurons, mainly of small size, are situated bilaterally in the caudal part of the nucleus. Anterograde tracing studies demonstrated the existence of SC projections to neurons in 5me. The SC fibers contact 'en passant' small as well as large cell bodies of 5me neurons. These pathways suggested a role of the 5me neurons in oculomotor control and associated oro-facial functions. PMID- 11044577 TI - Efficacy of intracochlear administration of betamethasone on peripheral vestibular disorder in the guinea pig. AB - We evaluated the effect of steroid hormone on vestibular function in a guinea pig of peripheral vestibular disorder. The right lateral semicircular canal was surgically damaged, and after surgery, animals were treated with 0.1 mg/ml of betamethasone in saline, 1 mg/ml of betamethasone in saline, or saline only, which was administrated directly into the scala tympani by osmotic pump. Rotation tests were performed, and the post-rotatory nystagmus (PRN) ratio (PRN number after counterclockwise rotation/PRN number after clockwise rotation) was calculated. The PRN ratio was recovered to normal at 5 days after treatment in the betamethasone administrated groups, but it did not recover to normal until 14 days after treatment in the saline administrated group. Results indicate that in the vestibular periphery steroid hormones may play an important role in recover of vestibular function. PMID- 11044579 TI - Neuroprotective effect of low dose riluzole in gerbil model of transient global ischemia. AB - Riluzole is a neuroprotective agent the efficacy of which was proven in amyotrophic lateral sclerosis in human and in animal models of cerebral ischemia. However, the dosage used in animal experiments was much higher than that in human. We investigated the efficacy of low dose riluzole, which was similar to the dose used in human trials, in animal model of global ischemia. Global ischemia was induced in male Mongolian gerbils for 5min under monitoring of rectal temperature. Riluzole (0.8 mg/kg) were injected intraperitoneally 30min before ischemia. Seven days after ischemia, animals were decapitated and surviving nerve cells in hippocampal CA1 area were quantified. The number of surviving cells was compared between in riluzole-treated and control groups and the former showed statistically significant better survivals than the latter (P<0.001). PMID- 11044578 TI - Androgen receptor mRNA expression in the human hippocampus. AB - The androgen receptor (AR) plays a central role in mediating androgen action. Since the hippocampus is a target of steroid modulation, we studied the expression of AR mRNAs in hippocampal tissue specimens from patients undergoing epilepsy surgery (n=42). AR mRNA expression was in the same order of magnitude than in prostate tissue, known for its high expression of AR. AR mRNA concentrations showed no significant difference in AR mRNA expression between men (49.3+/-8.0 arbitrary units (aU); mean+/-SEM) and women (54.3+/-11.2 aU) and no sex-specific hippocampal lateralization pattern was observed. No relationship could be detected between duration of epilepsy, individual seizure frequency, age of the patients and the expression levels of AR. The high expression of AR in the hippocampus suggests that this human brain area is an important target for androgen action. PMID- 11044580 TI - Elevation of the gamma-aminobutyric acid transaminase expression in the gerbil CA1 area after ischemia-reperfusion damage. AB - gamma-Aminobutyric acid-transaminase (GABA-T) plays an important role in the metabolism of GABA, particularly in the neurons or glial cells. The present study was undertaken to determine the alteration of GABA-T expression in the gerbil hippocampus after ischemia-reperfusion. In the sham, GABA-T(+) neurons were scattered in the hippocampus proper and dentate gyrus. The intensity of the GABA T immunoreactivity had nearly disappeared in the interneurons at 12 h after ischemia. In contrast, 24 h post-ischemia the dramatic augmentation of GABA-T immunoreactivity in the pyramidal cells was observed in the CA1 area but not in the CA2 or CA3 areas. Forty-eight hours after ischemia-reperfusion, its immunoreactivity was preserved in the CA1 neurons. These results suggest that the over-expression of GABA-T in the CA1 area may be related to delayed neuronal death after ischemia-reperfusion insult. PMID- 11044581 TI - Human leucocyte antigen-A2 increases risk of Alzheimer's disease but does not affect age of onset in a Scottish population. AB - The use of non-steroidal anti-inflammatory drugs has been associated with a reduced incidence of Alzheimer's disease (AD), suggesting that attenuation of the inflammatory response may be beneficial. Several, but not all, genetic association studies have shown human leucocyte antigen (HLA)-A2, a major histocompatibility complex class I antigen-binding transmembrane protein has an increased frequency in AD compared to controls, and in some reports is associated with a lowered age of onset. We further investigated the role of HLA-A2 in an independent sample of AD cases, including a large early onset cohort. The results of this current study and meta analysis of all studies available to date support previous evidence of an excess of HLA-A2 in AD, but found no evidence of a relationship with age of onset. PMID- 11044582 TI - Phase-dependent responses of Per1 and Per2 genes to a light-stimulus in the suprachiasmatic nucleus of the rat. AB - Single brief and discrete light treatments are sufficient to reset the overt mammalian rhythms of nocturnal rodents. In the present study, we examined the phase-dependent response of the mammalian clock genes, Per1 and Per2, to a brief strong light-stimulus (1000 lux) in the circadian oscillator center, the suprachiasmatic nucleus (SCN) of rats. Light-induced elevation of Per1 mRNA was observed through the subjective night (CT16, CT20 and CT0 (=CT24)) with a marked peak at the subjective dawn (CT0). However, the light influence was very limited for the induction of Per2; only weak elevation of Per2 mRNA was detected at CT16. The effect of light-stimulus on the Per1 gene was transient, and the effect was restricted to ventrolateral SCN neurons in both CT0 and CT16 after light exposure. Since it is known that these rats show a light-induced behavioral phase shift throughout the subjective night with being strongest at subjective dawn, the present results suggest that the transient induction of Per1 in ventrolateral SCN neurons is a critical step in the resetting of the biological clock to environmental light-dark schedule. PMID- 11044583 TI - Interleukin-6 expression in human multiple sclerosis lesions. AB - The present study investigated interleukin-6 (IL-6) expression in 36 multiple sclerosis (MS) cases by immunocytochemistry. The numbers of IL-6 expressing cells were correlated to the stage of demyelinating activity and the pattern of oligodendrocyte pathology. IL-6 positive cells were identified as macrophages and astrocytes by morphological criteria. Approximately 10-17% of the astrocytes and up to 2% of the macrophages within the lesion expressed IL-6. Highest numbers of IL-6 expressing cells were found in inactive demyelinating lesions. There was a significant increase of IL-6 positive cells in lesions with oligodendrocyte preservation, whereas absence of IL-6 expression correlated with oligodendrocyte loss. These observations indicate a possible important role for IL-6 in oligodendrocyte protection and survival in MS lesions. PMID- 11044584 TI - Properties and development of calcium currents in embryonic cockroach neurons. AB - In freshly dissociated neurons from embryonic cockroach (Periplaneta americana L.) brains, voltage-dependent calcium currents appear early in development (E14). Their intensity increases progressively during embryonic life until eclosion (E35). Their time course and voltage dependency are characteristic of high voltage activated (HVA) currents although a 10 mV shift of the I/V curve towards more negative potentials was observed between E18 and E23. Their sensitivity to omega-AgaTx-IVA and omega-CgTx-GVIA and insensitivity to both amiloride and isradipine indicate that the corresponding channels are of the P/Q and N types. These channels, as well as a small proportion of toxin-resistant (R) channels (about 20%), are blocked by mibefradil and verapamil. The physiological significance of these currents and their modifications during embryonic life is discussed. PMID- 11044585 TI - Dysfunction of M-channel enhances propagation of neuronal excitability in rat hippocampus monitored by multielectrode dish and microdialysis systems. AB - To explore the pathogenesis of benign familial neonatal convulsions (BFNC), we determined effects of KCNQ-related M-channels (KCNQ-channels) on hippocampal glutamate (Glu) and gamma-aminobutyric acid (GABA) releases using microdialysis, and propagation of evoked field-potentials (FP) using multielectrode (64-ch)-dish system as two-dimensional monitoring. KCNQ-channel inhibitor, Dup996, enhanced hippocampal K(+)-evoked Glu and GABA releases without affecting basal releases of them. Dup996 unaffected FP-amplitude, but enhanced FP-propagation. The GABA(A) receptor antagonist, bicuculline, enhanced the stimulatory effects of Dup996 on FP-propagation, however, this stimulatory effects of Dup996 were abolished by the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)/glutamate receptor antagonist, DNQX. These results suggest that the occurrence of BFNC cannot be produced by KCNQ-channel dysfunction alone, but by reciprocal action between impaired KCNQ-channel and other unknown elements (possibly dysfunction of inhibitory neurotransmission system). PMID- 11044586 TI - Nerve growth factor (NGF) and NGF mRNA change in rat uterus during pregnancy. AB - During pregnancy, the uterus undergoes a profound sympathetic denervation. To explore whether this is associated with changes in neurotrophic factors, we assayed nerve growth factor (NGF) and NGF mRNA in the uterus of non-pregnant and pregnant rats. In the uterine horn, the concentration of NGF and its mRNA decreased during middle and late pregnancy. However, when values were corrected for the increase of uterine weight and total RNA yield during pregnancy, NGF content and mRNA per horn increased during middle and late pregnancy. Similar, but less pronounced, changes were observed in the cervix. By seven days postpartum, both parameters returned to near normal. PMID- 11044587 TI - Promoter polymorphism of the 5-HT transporter and Alzheimer's disease. AB - The role of the deletion/insertion polymorphism within the promoter region of the serotonin transporter gene (5-HTT) is under discussion as a potential genetic risk factor for Alzheimers's disease (AD). Here we report significant differences in the allelic distribution of this polymorphism with a higher frequency of the short variant allele in AD patients when compared to controls. This difference was independent of the apolipoproteinE genotype. Thus, our study supports the notion that genetic alterations in the serontonergic neurotransmitter system may be involved in the etiopathogenesis of AD. However, given the reported negative findings, we are presently trying to identify diagnostic subgroups for which the 5-HTT promoter polymorphism represents a susceptibility locus. PMID- 11044588 TI - Dr. Otto Soltmann (1876) on development of the motor cortex and recovery after its removal in infancy. AB - In 1870, Fritsch and Hitzig demonstrated that dogs have a motor cortex. In a chapter published 6 years later, Otto Soltmann studied the functional development of the motor cortex, which he believed functioned in willed movement. He was the first to show that the dog's motor cortex becomes electrically excitable at about 10 days of age, with the contralateral forepaw area appearing first. He also studied the effects of ablating the cortical motor regions unilaterally and bilaterally, and encountered a remarkable degree of sparing of function in his animals operated on as newborns, but not in older-operated dogs. Soltmann turned to the theory of functional take-over (vicariation) to account for the absence of deficits in his young animals. He was especially intrigued by the fact that electrical stimulation of a healthy motor cortex could produce bilateral matched movements, but only in dogs that sustained opposite motor cortex lesions very early in life. PMID- 11044589 TI - Roles of molecular chaperones in the nervous system. AB - Heat shock proteins (HSPs) are induced not only by heat shock but also by various other environmental stresses. HSPs such as Hsp90, Hsp70, Hsp60, Hsp40 and Hsp28 are also expressed constitutively at normal growth temperatures and have basic and indispensable functions in the life cycle of proteins as molecular chaperones, as well as playing a role in protecting cells from deleterious stresses. Recently, Hsc70 and Hsp40 were found to be localized to the synapse in the mammalian central nervous system, indicating a synaptic role for these HSPs. Molecular chaperones are able to inhibit the aggregation of partially denatured proteins and refold them. In addition, molecular chaperones, especially Hsp70, protect the brain and heart from severe ischemia. In these respects, there are expectations for the use of molecular chaperones for protection against and therapeutic treatment of inherited diseases caused by protein misfolding. In this study, we review Hsp70 and Hsp40, and refer to the roles of these molecules in the synapse and cytoprotective functions of HSPs in stress tolerance and neurodegenerative diseases. PMID- 11044590 TI - Effects of neonatal cocaine treatment and gender on opioid agonist-stimulated [(35)S]GTP gamma S binding in the striatum and nucleus accumbens. AB - Prenatal cocaine exposure increases mu-opioid receptor binding in dopaminergic terminal areas and enhances behavioral responsiveness to mu-opioid agonists. We investigated the influence of early postnatal cocaine treatment on in vitro mu- and delta-opioid receptor activation in male and female weanling rats. Pups received subcutaneous injections of either 20 mg/kg cocaine HCl or saline once daily on postnatal days 1 through 5. On postnatal day 25, animals were decapitated and their brains were removed and frozen for later sectioning. Opioid receptor activation was assessed in the striatum and the shell of the nucleus accumbens by autoradiographic analysis of agonist-stimulated [(35)S]GTP gamma S binding. Brain sections were incubated in the presence of [(35)S]GTP gamma S, GDP, and either the mu-opioid agonist [D-Ala(2)-N-MePhe(4)-Gly(5)-ol]enkephalin (DAMGO) or the delta-opioid agonist D-Pen(2)-D-Pen(5)-enkephalin (DPDPE). Baseline binding was assessed in the absence of agonist, and nonspecific binding was determined by the addition of unlabeled GTP gamma S. Film images were quantified using brain mash-calibrated [(14)C] standards. Neonatal cocaine treatment had no effect on either baseline or agonist-stimulated [(35)S]GTP gamma S binding. However, males exhibited significantly greater activation than females of delta-opioid receptors in both striatum and accumbens shell, regardless of neonatal treatment. These findings indicate a gender difference in delta-opioid receptor function that could mediate behavioral differences in response to opioid agonists. PMID- 11044591 TI - Trypanosoma cruzi infection and the rat central nervous system: proliferation of parasites in astrocytes and the brain reaction to parasitism. AB - Chagas' disease, caused by the protozoan Trypanosoma cruzi, is characterized by an acute phase in which parasites circulate in the blood and proliferate in several cell types, especially muscle cells. A life-long chronic phase follows the acute phase. In young patients, the acute phase is more severe, and meningoencephalitis frequently occurs in children before 2 years of age. Parasites have been rarely observed in neurons but their presence inside glial cells has been reported without characterization of the glial cell type. The cells involved in the brain reaction to the parasites and the time course of this reaction remain to be studied. Therefore, using suckling and juvenile rats and different T. cruzi populations, we aimed at determining the brain target for parasite proliferation and the cells involved in the brain reaction. Around the middle of the acute phase, histological and ultrastructural findings indicated that T. cruzi proliferates in astrocytes, forming nests devoid of enclosing membrane as described for non-glial cells. The brain nodular reaction comprised astrocytes, microglia, macrophages and neutrophils. Resting microglia was devoid of parasites in contrast to macrophages and neutrophils that probably participate in parasite removal. Suckling animals were significantly more affected, the numbers of nests and nodules varying with inoculum size. Histoquantitative analysis showed higher number of nests at the parasitemic peak (day 13) and drastic fall at day 20 post-inoculation. The highest number of nodules occurred at day 20 with drastic reduction at day 30. Recovery from histopathological alterations occurred even in surviving younger animals. PMID- 11044592 TI - Collateral projection of substance P receptor expressing neurons in the medullary dorsal horn to bilateral parabrachial nuclei of the rat. AB - Employing a combination of fluorescent retrograde double labeling and immunofluorescence histochemistry for substance P receptor (SPR), we examined the collateral projection from single SPR-like immunoreactive neurons in the medullary dorsal horn (caudal subnucleus of the spinal trigeminal nucleus) to bilateral parabrachial nuclei in the rat. After injection of fast blue (FB) or diamidino yellow (DY) into the right or left parabrachial nucleus, respectively, single-labeled FB or DY neurons and double-labeled FB/DY neurons were observed mainly bilaterally in laminae I and II of the medullary dorsal horn. Some of the single-labeled FB or DY and double-labeled FB/DY neurons showed SPR-like immunoreactivity, especially in lamina I. In lamina I of left medullary dorsal horn which is ipsilateral to the DY injection into the PBN, the percentages of double-labeled FB/DY neurons to the total number of FB- or DY-labeled neurons were 34.0% or 20.2%, triple-labeled FB/DY/SPR neurons to the total number of FB/DY double-labeled or SPR-like immunoreactive neurons were 22.0% or 2.4%, respectively. In lamina I of the right medullary dorsal horn which is ipsilateral to the FB injection into the PBN, the percentages of double-labeled FB/DY neurons to the total number of FB- or DY-labeled neurons were 12.9% or 59.3%, triple labeled FB/DY/SPR neurons to the total number of double-labeled FB/DY or SPR-like immunoreactive neurons were 24. 6% or 3.9%, respectively. The results suggest that some of the single SPR expressing neurons in the medullary dorsal horn might be innervated by substance P containing primary afferent fibers and transmit sensory information diffusely to bilateral parabrachial nuclei by way of their axonal collaterals. PMID- 11044593 TI - Effect of norepinephrine receptors on trigeminal rhythm generation in newborn rats. AB - N-methyl-D,L-aspartate acid and bicuculline are required to enhance the trigeminal rhythmic activities in an in vitro isolated brainstem block preparation. In this study, we analyzed the effect of norepinephrine on the trigeminal neural circuit underlying rhythmic jaw movements. Rhythmic trigeminal activity is observed in brainstem preparations (inferior colliculus to obex) only following blockade of alpha(2)-adrenoceptors with idazoxan. This observation, combined with the inhibition of rhythm by alpha(2)-adrenoceptor agonists suggests endogenous alpha(2)-adrenoceptor mediated inhibition of trigeminal networks. A complex noradrenergic modulation of trigeminal systems is further supported by the prazosin-sensitive potentiation of rhythm by bath application of the alpha(1) adrenoceptor agonist phenylephrine. PMID- 11044594 TI - Large cortical lesions produce enduring forelimb placing deficits in un-treated rats and treatment with NMDA antagonists or anti-oxidant drugs induces behavioral recovery. AB - Previous studies have utilized a lesion model of cortical injury that produces transient behavioral impairments to investigate the recovery of function process. To better understand the recovery process, it would be beneficial to use a lesion model that produces more severe, enduring, behavioral impairments. The purpose of experiment 1 was to validate whether large lesions of the sensorimotor cortex (SMC), which included the rostral forelimb and caudal forelimb regions, produced enduring behavioral deficits. Rats were given large unilateral electrolytic lesions of the SMC, administered either the N-methyl-D-aspartate (NMDA) antagonist, MK-801 or saline 16 h after injury, and tested on a battery of behavioral tests. Enduring behavioral deficits were observed, for at least 6 months, on two tests of forelimb placing while transient deficits were observed on the foot-fault and somatosensory neutralization tests. Administration of MK 801 facilitated recovery on the somatosensory neutralization test; however, it did not induce recovery on either forelimb placing test. A second experiment was performed to determine if earlier administration of MK-801, the NMDA antagonist magnesium chloride (MgCl(2)), or the anti-oxidant N-tert-butyl-alpha phenylnitrone (PBN) could induce behavioral recovery in this chronic model. Treatment with these drugs induced behavioral recovery on the forelimb placing tests, whereas, the saline-treated rats did not show any signs of behavioral recovery for at least 3 months. Anatomical analysis of the striatum showed that MK-801 and MgCl(2) but not PBN reduced the extent of lesion-induced striatal atrophy. These results suggest that administration of MK-801, MgCl(2), or PBN shortly after cortical injury can induce recovery of function when recovery is otherwise not expected in un-treated rats. PMID- 11044595 TI - Methylphenidate affects striatal dopamine differently in an animal model for attention-deficit/hyperactivity disorder--the spontaneously hypertensive rat. AB - The spontaneously hypertensive rat (SHR) is used as a model for attention deficit/hyperactivity disorder (ADHD) because it has behavioural characteristics (hyperactivity, impulsiveness, poorly sustained attention) similar to those of ADHD. ADHD children have been shown to have reduced striatal activation in certain tasks. SHR have reduced striatal dopamine release in response to electrical stimulation. The present study set out to investigate possible long term effects of methylphenidate treatment on dopaminergic function in striatal slices of SHR compared to their normotensive Wistar-Kyoto (WKY) control rats. Methylphenidate treatment (3 mg/kg daily for 14 days) did not normalize the decreased electrically-stimulated release of [(3)H]dopamine from SHR caudate putamen slices nor did it affect postsynaptic D(2) receptor function. However, the second electrical stimulus caused a relatively greater release of [(3)H]dopamine from caudate-putamen slices of methylphenidate-treated SHR than from vehicle-treated SHR, suggesting that presynaptic mechanisms controlling dopamine release had been altered. Interestingly, [(3)H]dopamine release from WKY caudate-putamen slices in response to D(2) autoreceptor blockade by the antagonist, sulpiride, was selectively increased by methylphenidate treatment. This effect was not seen in SHR possibly because D(2) autoreceptor function had already been up-regulated. The results show that methylphenidate is unable to enhance D(2) autoreceptor function in SHR. PMID- 11044596 TI - Permanent and transitory morphometric changes of NADPH-diaphorase-containing neurons in the rat visual cortex after early malnutrition. AB - We investigated the histochemical positivity to NADPH-diaphorase, which reveals nitric oxide synthase activity, in area 17 of rats malnourished early in life, both in the post-weaning period (group M1), and in adulthood after nutritional recovering (group M2). Control pups (C1 and C2 groups) received ad libitum after weaning the same diets as their mothers. Rats of group M2 were nutritionally recovered by receiving the control diet from post-natal day 42 until adulthood. Aldehyde-fixed sections (200-microm thick) through area 17 were processed for NADPH-diaphorase histochemistry following the malic enzyme indirect method. The features of NADPH-diaphorase-containing neurons of area 17 of malnourished young (M1) and adult (M2) rats were analyzed quantitatively in comparison to the matched groups C1 and C2. Permanent changes, represented by increase in the density and dendritic field areas of NADPH-diaphorase-positive cells, and transitory ones, represented by decreased values of soma areas, were observed in area 17 of the M1 and M2 cases. However, some other features, such as dendritic branch angle and number of dendrites per cell in the gray matter, remained unchanged after malnutrition. Thus, the findings indicate a possible relationship between early malnutrition and alterations in nitric oxide synthase-containing cells in the visual cortex. Physiological implications of these data may be related to synaptic plasticity and refinement of developmental brain circuits. PMID- 11044597 TI - Differential effects of K(+) channel blockers on frequency-dependent action potential broadening in supraoptic neurons. AB - Recordings were made from magnocellular neuroendocrine cells dissociated from the supraoptic nucleus of the adult guinea pig to determine the role of voltage gated K(+) channels in controlling the duration of action potentials and in mediating frequency-dependent action potential broadening exhibited by these neurons. The K(+) channel blockers charybdotoxin (ChTx), tetraethylammonium (TEA), and 4 aminopyridine (4-AP) increased the duration of individual action potentials indicating that multiple types of K(+) channel are important in controlling action potential duration. The effect of these K(+) channel blockers was almost completely reversed by simultaneous blockade of voltage gated Ca(2+) channels with Cd(2+). Frequency-dependent action potential broadening was exhibited by these neurons during trains of action potentials elicited by membrane depolarizing current pulses presented at 10 Hz but not at 1 Hz. 4-AP but not ChTx or TEA inhibited frequency-dependent action potential broadening indicating that frequency-dependent action potential broadening is dependent on increasing steady state inactivation of A-type K(+) channels (which are blocked by 4-AP). A model of differential contributions of voltage gated K(+) channels and voltage gated Ca(2+) channels to frequency-dependent action potential broadening, in which an increase of Ca(2+) current during each successive action potential is permitted as a result of the increasing steady-state inactivation of A-type K(+) channels, is presented. PMID- 11044598 TI - Effects of combined administration of zonisamide and valproic acid or phenytoin to nitric oxide production, monoamines and zonisamide concentrations in the brain of seizure-susceptible EL mice. AB - This study was undertaken to elucidate the anticonvulsive effects of zonisamide (ZNS: 25, 50, and 75 mg/kg, intraperitoneal [i.p.]), which was coadministered with valproic acid (VPA: 25, 50, and 100 mg/kg, i.p.), or phenytoin (PHT: 10, 25, and 50 mg/kg, i.p.) to ZNS concentration, nitric oxide metabolites (NOx levels), and monoamines in the brain of the EL mouse, a strain highly susceptible to seizures. NOx levels were obtained from measuring of combined level of nitrite plus nitrate. Coadministration of ZNS with VPA or PHT suppressed convulsive seizures more effectively than with treatment of ZNS alone. Both serum and brain concentrations of ZNS tended to increase as the dose of VPA or PHT was increased. While coadministrations of ZNS (75 mg/kg) and VPA or PHT at any dose did not change brain and serum NOx levels, those altered brain monoamine contents. These results suggested that anticonvulsive effect of coadministrations of ZNS and VPA or PHT were caused by changes of monoamines rather than changes of NO metabolites. PMID- 11044599 TI - Basal levels and alcohol-induced changes in nociceptin/orphanin FQ, dynorphin, and enkephalin levels in C57BL/6J mice. AB - In order to investigate the involvement of the opioid and nociceptin/orphanin FQ (N/OFQ) system in alcohol drinking behaviour, N/OFQ and the opioid peptides dynorphin B (DYNB) and Met-enkephalin-Arg(6) Phe(7) (MEAP) were examined in the alcohol-preferring C57BL/6J mice. Basal peptide levels were compared in the brain and the pituitary gland with basal levels in the alcohol-avoiding DBA/2J mice. Furthermore, the effects of chronic alcohol self-administration on peptides were studied in the C57BL/6J mice. Compared to the DBA/2J mice, C57BL/6J mice had low immunoreactive (ir) levels of DYNB and MEAP in the nucleus accumbens, the hippocampus, and the substantia nigra, low ir-DYNB levels in the striatum and low ir-MEAP levels in the frontal cortex. Higher ir-DYNB levels in the pituitary gland and in the periaqueductal gray (PAG) and higher ir-N/OFQ levels in the frontal cortex and in the hippocampus were detected in C57BL/6J mice compared to the DBA/2J mice. After 4 weeks of voluntary alcohol consumption, only minor changes in steady-state peptide levels were identified. However, 5 days after the alcohol-drinking period, lower levels of all peptides were detected in the ventral tegmental area and ir-DYNB levels were also lower in the amygdala and in the substantia nigra. Twenty-one days after cessation of alcohol self administration, the opioid peptides in alcohol-consuming C57BL/6J mice were lower in the PAG, the N/OFQ was lower in the frontal cortex and DYNB was higher in the amygdala and substantia nigra as compared to control C57BL/6J mice. This study demonstrates strain differences between C57BL/6J mice and DBA/2J mice that could contribute to divergent drug-taking behaviour, and it also demonstrates time- and structure-specific changes in neuropeptide levels after alcohol self administration in the C57BL/6J mice. PMID- 11044600 TI - The effect of callosotomy on testicular steroidogenesis in hemiorchidectomized rats: a pituitary-independent regulatory mechanism. AB - In recent years, increasing number of data indicate that cerebral structures exert a direct, pituitary-independent, neural regulatory action on the endocrine glands. In addition, both experimental and clinical observations indicate functional asymmetry of the control system. Therefore, the objective of the present study was to study the effect of callosotomy on testicular steroidogenesis and serum gonadotrop concentrations in rats subjected to left- or right-sided orchidectomy. In animals underwent callosotomy plus left-sided orchidectomy the basal testosterone secretion in vitro of the remaining (right) testis was significantly higher than that of intact controls, and of rats subjected to sham surgery plus left orchidectomy. In contrast, either sham operation or callosotomy plus right-sided orchidectomy did not interfere with testicular steroidogenesis. Sham surgery or callosotomy plus left orchidectomy induced a significant rise in serum follicle-stimulating hormone concentration while right orchidectomy combined either with sham surgery or callosotomy did not alter this parameter. There was no statistically significant difference between experimental groups in serum testosterone and luteinizing hormone concentrations. The results indicate the involvement of the corpus callosum in a pituitary independent neural control of testicular steroidogenesis. The data further suggest a different response in steroidogenesis of the left and the right testis following hemicastration and callosotomy. PMID- 11044601 TI - Static contraction causes a reflex-induced release of arginine vasopressin in anesthetized cats. AB - We tested the hypothesis that brief static contraction of the triceps surae muscle causes reflex-induced increases in plasma arginine vasopressin (AVP) in anesthetized cats. Arterial blood samples, for measurement of plasma AVP, were taken before and after 30 s of electrically stimulated static contraction performed at a low intensity (<20% of maximal; n = 5), a high intensity (>70% of maximal; n = 7), and a high intensity after denervation of the triceps surae (n = 5). The low intensity contraction protocol was repeated during alpha-adrenergic blockade (n = 7) to minimize potential baroreflex-induced inhibition of AVP release. Passive stretch of the triceps surae was conducted (n = 5) to determine effects of muscle mechanoreceptor stimulation on the release of AVP. Low intensity contraction had no effect on plasma AVP. During alpha-adrenergic blockade, this same contraction intensity caused this peptide to increase from 12.8 +/- 2.1 to 17.7 +/- 2.6 pg/ml. High intensity contraction caused an increase in AVP (13.2 +/- 3.5 to 26.1 +/- 6.6 pg/ml) that was abolished by denervation (14.4 +/- 3. 7 vs. 17.1 +/- 6.6 pg/ml). Passive stretch had no effect on plasma AVP. These findings suggest that brief static contraction causes increases in plasma AVP that are reflex in nature, intensity dependent, opposed by the arterial baroreflex, and probably unrelated to muscle mechanoreceptor activation. PMID- 11044602 TI - Retinal projections to the pregeniculate nucleus in the hemispherectomized monkey. AB - Intraocular injections of tritiated proline were used to test the hypothesis that unilateral removal of all visual cortical areas results in increased distribution of retinal terminals in the pregeniculate nucleus (PGN) of the thalamus in monkeys. Following hemispherectomy, retinal input to the ipsilateral PGN was reduced by an average of 18.5% when compared to its contralateral homologue, which corresponded to the reduction in nuclear volume (19.3%). Our results show that removal of cortical afferents to the external layer of the PGN does not induce invasion of retinal projections into this region of the nucleus. PMID- 11044603 TI - Zebrafish Dkk1, induced by the pre-MBT Wnt signaling, is secreted from the prechordal plate and patterns the anterior neural plate. AB - mRNA injection into the ventral blastomeres of Xenopus embryos of mRNA encoding Wnt pathway genes induces a secondary axis with complete head structures. To identify target genes of the pre-MBT dorsalization pathway that might be responsible for head formation in zebrafish, we have cloned zebrafish dickkopf1 (dkk1), which is expressed in tissues implicated in head patterning. We found that dkk1 blocks the post-MBT Wnt signaling and dkk1 is a target of the pre-MBT Wnt signaling. Dkk1 overexpression in the prechordal plate suggests that Dkk1, secreted from the prechordal plate, expands the forebrain at the expense of the midbrain in the anterior neural plate. Furthermore, dkk1 acts in parallel to the homeobox gene bozozok and bozozok is required for the maintenance of dkk1 expression. The nodal gene squint is also required for the maintenance of dkk1 expression. Among the mutually dependent target genes of the pre-MBT Wnt signaling, dkk1 plays an important role in patterning the anterior head of zebrafish. PMID- 11044604 TI - Neural hyperplasia induced by RNA interference with m4/malpha gene activity. AB - The E(spl) complex (E(spl)-C) contains three different classes of genes that are downstream of Notch signaling. The bHLH genes mediate the Notch signal by repressing proneural gene activity, for example during the singularization of mechanosensory organ precursor cells (SOPs). Genes of the second class, the E(spl) m4/malpha family, antagonize this process if overexpressed. Here we show that this is based on dominant-negative effects since RNA interference gives neurogenic phenotypes indistinguishable from E(spl)-C mutations. Furthermore, a third member of the m4/malpha gene family, named bbu/tom, behaves differently with respect to RNA expression patterns, its regulation by Notch signaling and loss of function phenotypes. PMID- 11044605 TI - c-Jun (AP-1) activates BMP-4 transcription in Xenopus embryos. AB - Zygotic expression of the BMP-4 gene in Xenopus embryos is regulated by an auto regulatory loop. Since AP-1 is known as a mediator of auto-regulatory loops both in the case of the Drosophila dpp and the mammalian TGF-beta genes, we have analysed the potential of Xenopus c-Jun (AP-1) as a mediator of BMP-4 expression during Xenopus development. RNA injection experiments revealed that both heteromeric c-Fos/c-Jun and homodimeric c-Jun/c-Jun strongly activate BMP-4 transcription, whereas BMP signaling was found to activate the Xenopus c-Jun gene only at a rather low extent. In addition, the lack of zygotic c-Jun transcripts until the end of gastrulation should exclude a role of AP-1 in the activation and the early expression of BMP-4 during gastrulation in vivo. However, at later stages of Xenopus development, we find a spatial overlap of c-Jun and BMP-4 transcripts which suggests that AP-1 might serve as an additional activatory component for the auto-regulation of BMP-4. Promoter/reporter and gel mobility shift assays demonstrate multiple responsive sites for AP-1 in the 5' flanking region and two in the second intron of the BMP-4 gene. We further demonstrate that AP-1 acts independently of Xvent-2 which has recently been shown to mediate the early expression of BMP-4 in gastrula stage embryos. PMID- 11044606 TI - Identification of RALDH-3, a novel retinaldehyde dehydrogenase, expressed in the ventral region of the retina. AB - In the developing retina, a retinoic acid (RA) gradient along the dorso-ventral axis is believed to be a prerequisite for the establishment of dorso-ventral asymmetry. This RA gradient is thought to result from the asymmetrical distribution of RA-generating aldehyde dehydrogenases along the dorso-ventral axis. Here, we identified a novel aldehyde dehydrogenase specifically expressed in the chick ventral retina, using restriction landmark cDNA scanning (RLCS). Since this molecule showed enzymatic activity to produce RA from retinaldehyde, we designated it retinaldehyde dehydrogenase 3 (RALDH-3). Structural similarity suggested that RALDH-3 is the orthologue of human aldehyde dehydrogenase 6. We also isolated RALDH-1 which is expressed in the chick dorsal retina and implicated in RA formation. Raldh-3 was preferentially expressed first in the surface ectoderm overlying the ventral portion of the prospective eye region and then in the ventral retina, earlier than Raldh-1 in chick and mouse embryos. High level expression of Raldh-3 was also observed in the nasal region. In addition, we found that Pax6 mutants are devoid of Raldh-3 expression. These results suggested that Raldh-3 is the key enzyme in the formation of an RA gradient along the dorso-ventral axis during the early eye development, and also in the development of the olfactory system. PMID- 11044607 TI - Large-scale screen for genes involved in gonad development. AB - The vertebrate gonad develops from the intermediate mesoderm as an initially bipotential organ anlage, the genital ridge. In mammals, Sry acts as a genetic switch towards testis development. Sox9 has been shown to act downstream of Sry in testis development, while Dax1 appears to counteract Sry. Few more genes have been implicated in early gonad development. However, the genetic networks controlling early differentiation events in testis and ovary are still far from being understood. In order to provide a broader basis for the molecular analysis of gonad development, high-throughput gene expression analysis was utilized to identify genes specifically expressed in the gonad. In total, among 138 genes isolated which showed tissue specific expression in the embryo, 79 were detected in the developing gonad or sex ducts. Twenty-seven have not been functionally described before, while 40 represent known genes and 12 are putative mouse orthologues. Forty-five of the latter two groups (86%) have not been described previously in the fetal gonad. In addition, 21 of the gonad specific genes showed sex-dimorphic expression suggesting a role in sex determination and/or gonad differentiation. Eighteen of the latter (86%) have not been described previously in the fetal gonad. In total we provide new data on 72 genes which may play a role in gonad or sex duct development and/or sex determination. Thus we have generated a large gene resource for the investigation of these processes, and demonstrate the suitability of high-throughput gene expression screening for the genetic analysis of organogenesis. PMID- 11044608 TI - An autoregulatory function of Dfos during Drosophila endoderm induction. AB - The endoderm of Drosophila is patterned during embryogenesis by an inductive cascade emanating from the adhering mesoderm. An immediate-early endodermal target gene of this induction is Dfos whose expression is upregulated in the middle midgut by Dpp signalling. Previous evidence based on a dominant-negative Dfos construct indicated that Dfos may cooperate with Dpp signalling to induce the HOX gene labial, the ultimate target gene of the inductive cascade. Here, we examine kayak mutants that lack Dfos to establish that Dfos is indeed required for labial induction. We provide evidence that Dfos acts through a CRE-like sequence, previously identified to be a target for signalling by Dpp and by the Epidermal growth factor receptor (Egfr) in the embryonic midgut. We show that Dfos expression is stimulated by Egfr signalling. Finally, we find that Dfos function is required for its own upregulation. Thus, endoderm induction is based on at least four tiers of positive autoregulatory feedback loops. PMID- 11044609 TI - Large-scale screen for genes controlling mammalian embryogenesis, using high throughput gene expression analysis in mouse embryos. AB - We have adapted the whole-mount in situ hybridization technique to perform high throughput gene expression analysis in mouse embryos. A large-scale screen for genes showing specific expression patterns in the mid-gestation embryo was carried out, and a large number of genes controlling development were isolated. From 35760 clones of a 9.5 d.p.c. cDNA library, a total of 5348 cDNAs, enriched for rare transcripts, were selected and analyzed by whole-mount in situ hybridization. Four hundred and twenty-eight clones revealed specific expression patterns in the 9.5 d.p.c. embryo. Of 361 tag-sequenced clones, 198 (55%) represent 154 known mouse genes. Thirty-nine (25%) of the known genes are involved in transcriptional regulation and 33 (21%) in inter- or intracellular signaling. A large number of these genes have been shown to play an important role in embryogenesis. Furthermore, 24 (16%) of the known genes are implicated in human disorders and three others altered in classical mouse mutations. Similar proportions of regulators of embryonic development and candidates for human disorders or mouse mutations are expected among the 163 new mouse genes isolated. Thus, high-throughput gene expression analysis is suitable for isolating regulators of embryonic development on a large-scale, and in the long term, for determining the molecular anatomy of the mouse embryo. This knowledge will provide a basis for the systematic investigation of pattern formation, tissue differentiation and organogenesis in mammals. PMID- 11044610 TI - Notch/Delta expression in the developing mouse lung. AB - Factors controlling the differentiation of the multipotent embryonic lung endoderm and mesoderm are poorly understood. Recent evidence that Delta-like 1 (Dll1) and other genes in the Notch/Delta signaling pathway are expressed in the embryonic mouse lung suggests that this pathway is important for cell fate decisions and/or the differentiation of lung cell types. Here, we report the localization of transcripts of several genes encoding members of the Notch/Delta pathway in the early mouse lung. Most genes are expressed in specific populations and so may contribute to cell diversification. PMID- 11044611 TI - Distinct expression of two types of Xenopus Patched genes during early embryogenesis and hindlimb development. AB - Patched (Ptc) is a putative twelve transmembrane domain protein that is both a Hedgehog (Hh) receptor and transcriptional target of Hh. In this study, we isolated Xenopus Ptc cDNAs, Ptc-1 and Ptc-2, and carried out comparative analyses on their expression patterns. The putative Ptc-2 protein has a long C-terminal extension that has similarities in both length and sequence to those of Ptc-1 proteins in mouse, chick and human. In both early embryogenesis and hindlimb development, Ptc-2 expression is restricted to cells that receive a Hh signal, a pattern similar to that of Gli-1. Ptc-1, however, shows a broader distribution, mainly non-overlapping with that of Ptc-2. Despite the difference in their expression patterns, both are induced in animal cap explants synergistically by Shh and Noggin, showing a conserved regulation in their activation mechanisms. PMID- 11044612 TI - Differential spatial distribution and temporal regulation of tissue inhibitor of metalloproteinase mRNA expression during rat central nervous system development. AB - The elucidation of the cellular and molecular mechanisms governing the maturation of the central nervous system (CNS) is rapidly emerging. Cell-cell and cell matrix interactions play critical roles in all phases of developmental tissue remodeling. Throughout development, an intricate balance between extracellular matrix synthesis and degradation is preserved by the opposing actions of matrix metalloproteinases (MMPs) and their specific inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). Although recent evidence suggests that TIMPs exert diverse cell biological functions distinct from their MMP-inhibitory activities, few studies have investigated MMP or TIMP expression during CNS development. The present report analyzes the mRNA expression of the four known TIMPs throughout the course of embryonic and postnatal rat CNS development. The results clearly demonstrate the unique spatial distribution and temporal regulation of TIMP expression and suggest a distinct role for each TIMP during CNS development. PMID- 11044613 TI - The murine A33 antigen is expressed at two distinct sites during development, the ICM of the blastocyst and the intestinal epithelium. AB - The expression pattern of the murine A33 antigen has been defined during development using wholemount immunohistochemistry. Two temporally and spatially distinct sites of expression were identified: the inner cell mass of the blastocyst and the endoderm cell layer of the intestinal tract where expression is initiated at E14.5 in the hindgut and subsequently extends throughout the length of the intestine. The onset of mA33 antigen expression in the gut occurs at the beginning of an extensive phase of cell movement involved in the conversion of the endoderm cell layer to a single cell layer of polarized epithelium. Expression of mA33 antigen is then maintained into adulthood, where it is a definitive marker of intestinal epithelium. PMID- 11044614 TI - Molecular cloning and expression analyses of mouse betaklotho, which encodes a novel Klotho family protein. AB - We report here the identification of mouse betaklotho (betakl), which encodes a type I membrane protein with high resemblance to Klotho (KL). Both betaKL and KL consist of two internal repeats with homology to family 1 glycosidases, while these essential glutamates for the enzymatic activities were not conserved. The identical pattern of substitution and variation in the substituted amino acids between these two proteins indicate that they likely to form a unique family within the glycosidase family 1 superfamily. During mouse embryonic development, strong betakl expression was detected in the yolk sac, gut, brown and white adipose tissues, liver and pancreas, and in the adult, predominantly in the liver and pancreas. Despite the high structural similarity between betaKL and KL, their expression profiles were considerably different and betakl expression was not induced in kl-deficient mouse mutants. PMID- 11044615 TI - Frizzled-4 expression during chick kidney development. AB - Wnts have been implicated in metanephric kidney development. To determine whether Frizzleds, the genes that encode Wnt receptors, are present at early stages of nephrogenesis, we examined the expression of several recently identified Frizzled genes in the chick by in situ hybridization. Here we report the cloning and characterization of chick Frizzled-4 (cFz-4), which we found to be expressed in the developing chick kidney. cFz-4 was first expressed in the pronephros caudal to the third somite at Hamburger and Hamilton stage 10. Its expression increased with maturation, becoming restricted to the newly induced glomeruli and tubules in the mesonephros and metanephros. Within the metanephros, cFz-4 and Wnt-4 expression patterns were similar, whereas Wnt-11 was expressed solely in the tips of the branching ureteric bud. cFz-4 expression was compared with that of known kidney markers. It preceded that of Lmx-1, but was similarly restricted to developing glomeruli and tubules. In contrast, Pax-2 expression and Lim 1/2 antibody labeling occurred in intermediate mesoderm caudal to the fifth somite in the early pronephros, and each persisted in both the tubules and nephric ducts throughout further development. PMID- 11044616 TI - Expression of the G protein-coupled receptor kinase 2 during early mouse embryogenesis. AB - Whilst several G protein-coupled receptors (GPCRs) have been shown to play important roles during development, little study has been carried out on the G protein-coupled receptor kinases (GRKs) that modulate their activity in embryos. Here, we have analyzed the expression of GRK2, the predominant GRK expressed during embryogenesis. We show that at early stages, the expression of GRK2 is restricted to populations of cells that are undifferentiated, multipotent and in many cases, migratory. As such, GRK2 transcripts were found in the early mesoderm and neural crest as they migrate from the primitive streak and the neural tube, respectively. In the limb bud, GRK2 transcripts were observed in cells of the progress zone and in the interdigital areas. At later stages, the expression in the heart is compatible with the phenotype observed in the GRK2 deficient mice. PMID- 11044617 TI - Expression of mouse HES-6, a new member of the Hairy/Enhancer of split family of bHLH transcription factors. AB - We studied the expression of mouse HES-6, a new member of the Hairy/Enhancer of split family of basic helix-loop-helix transcription factors. HES-6 is expressed in all neurogenic placodes and their derivatives and in the brain, where it is patterned along both the anteroposterior and dorsoventral axes. HES-6 is also expressed in the trunk, in the dorsal root ganglia and in the myotomes. In the limb buds HES-6 is expressed in skeletal muscle and presumptive tendons. PMID- 11044618 TI - Cloning and embryonic expression patterns of the zebrafish Runt domain genes, runxa and runxb. AB - We isolated zebrafish homologues of the Runt-related transcription factor gene family (Runx family), runxa and runxb, and analyzed their developmental expression patterns. The deduced amino acid sequence of Runxa was highly homologous to that of AML1 (also called CBFA2, PEBP2alphaB or Runx1), a critical regulator of mammalian hematopoiesis expressed in cells of the hematopoietic lineage as well as other tissues. During zebrafish development, the runxa gene was not expressed in hematopoietic tissues but in the olfactory placodes and cells attached to the otic vesicles. We identified three kinds of runxb transcripts, which encoded two types of proteins with different N-terminal regions. The Runxb proteins were highly similar to AML2 (CBFA3, PEBP2alphaC or Runx3). The expression sites of the shared region of runxb mRNAs during development were the trigeminal ganglions, dorsal neurons of the neural tube and the lateral line primordia. These findings show that expression patterns of the zebrafish Runx genes are distinct from that of the mammalian genes. PMID- 11044619 TI - Differential expression of two glucuronyltransferases synthesizing HNK-1 carbohydrate epitope in the sublineages of the rat myogenic progenitors. AB - HNK-1 epitope is a cell-surface carbohydrate mediating various cell-cell or cell substrate interactions. We found HNK-1 epitope in longitudinally arrayed fibers in the subpopulation of the epaxial myotome, and hypaxial myoblasts migrating into the limb bud in the rat embryo. We next investigated the expression patterns of genes encoding two glucuronyltransferases (GlcAT-P, GlcAT-D) and sulfotransferase (Sul-T), which are required for biosynthesis of HNK-1 epitope. GlcAT-P gene was expressed in the non-migrating longitudinal fibers, whereas GlcAT-D gene was expressed in the migrating myoblasts in the limb bud. Sul-T gene expression was ubiquitously observed in all these myogenic populations. Thus, differential expression of GlcAT genes may relate to the epaxial/hypaxial or migrating/non-migrating myoblast lineages. PMID- 11044620 TI - Expression of three zebrafish Six4 genes in the cranial sensory placodes and the developing somites. AB - The homeobox gene Six4/AREC3 is a member of the vertebrate Six family of transcription factor genes. In this study we describe the cloning and expression of three zebrafish homologues of Six4/AREC3, six4.1, six4.2 and six4.3. These zebrafish Six4 proteins show high homology to the mammalian Six4/AREC3 proteins in the most C-terminal region in addition to the Six domain and homeodomain. Whole-mount in situ hybridization showed that six4.1 is expressed in the presumptive cranial placodal precursor cells, and later in the olfactory, otic and lateral line placodes. six4.2 is expressed in the presomitic mesoderm, somites and pectoral fin bud. six4.3 appears to be a unique member of the Six4 proteins and is expressed ubiquitously at gastrulation and later in the tectum. PMID- 11044621 TI - Expression analysis of the chicken homologue of CITED2 during early stages of embryonic development. AB - Members of the Cited family are nuclear transactivators which bind to the coactivators p300 and CBP. While Cited1 also binds to the TGFbeta signal transducer Smad4, this has not been shown for Cited2. We isolated a chicken homologue of Cited2 from a HH stage 3-6 cDNA library and examined its expression pattern during early stages of embryonic development by whole-mount in situ hybridization. CITED2 expression is detectable in the epiblast as early as stage XI. From HH stage 2 onwards CITED2 is expressed in an anterior domain in the elongating primitive streak in cells which are fated to become heart. During gastrulation the expression pattern is highly dynamic and transiently displays left-right asymmetry with stronger expression on the right side. CITED2 expression appears at multiple sites of forming mesodermal structures. Most prominently, CITED2 is expressed in presomitic and lateral plate mesoderm, in the headfold (future forebrain), the head mesoderm, the pharyngeal floor, the ventral blood islands, somitomeres and the intermediate mesoderm which gives rise to the kidney anlagen. PMID- 11044622 TI - A new murine zinc finger gene, Opr. AB - Here we report a novel murine zinc-finger gene, Opr, belonging to the opa/Zic family. Opr is expressed in the entire embryonic ectoderm before gastrulation, but gradually restricted to the anterior part from the mid to late streak stage. At the beginning of neural induction, Opr is expressed throughout the anterior neural plate, but is soon restricted to the neural ridge. After neural tube closure, its expression is maintained in the dorsal part of the neural tube, except for the roof of the telencephalon. Opr is also expressed in somites and limbs. PMID- 11044623 TI - Regulation of Hoxb3 expression in the hindbrain and pharyngeal arches by rae28, a member of the mammalian Polycomb group of genes. AB - During animal development, Hox genes are expressed in characteristic, spatially restricted patterns and specify regional identities along the anterior-posterior (A-P) axis. Polycomb group (PcG) proteins in Drosophila repress Hox expression and maintain the expression patterns during development. Mice deficient for homologues of the Drosophila PcG genes, such as M33, bmi1, mel18, rae28 and eed, show altered Hox expression patterns. In this study, we examined the time course of Hoxb3 expression during late gastrulation and early segmentation of rae28 deficient mice. Hoxb3 was expressed ectopically in pharyngeal arch and hindbrain from embryonic day (E) 9.5 and 10.5, respectively. The anterior boundary of ectopic expression in the hindbrain extended gradually in the rostral direction as development proceeded from E10.5 to E12.5. Expression of kreisler and Krox20, which function as positive regulators of Hoxb3 expression, was not affected in rae28-deficient embryos. Analysis of a neural crest marker, p75, in rae28 deficient mice revealed that the neural crest cells begin to ectopically express Hoxb3 after leaving the hindbrain. Our results suggest that rae28 is not required for the establishment but maintenance of Hoxb3 expression. PMID- 11044624 TI - Similar expression and regulation of Gli2 and Gli3 in the chick limb bud. AB - Gli genes encode a family of zinc finger transcription factors that mediate signaling by Hedgehog proteins. We have cloned the chick Gli3 gene and studied its expression in developing chick limbs. Gli3 expression is highly similar to that of chick Gli2. Gli3 mRNA is evenly distributed in the early limb mesenchyme and subsequently downregulated in the posterior mesenchyme by the polarizing activity of Sonic hedgehog. At later stages, Gli3 is expressed in the distal limb mesenchyme. PMID- 11044625 TI - Analysis of HeyL expression in wild-type and Notch pathway mutant mouse embryos. AB - In vertebrates Notch signaling regulates cell fate decisions and boundary formation and it underlies several murine and human diseases. Gene targeting experiments point to key roles of Notch receptors, ligands, modulators and downstream targets in somitogenesis, neurogenesis and vascular development. Here we report the embryonic expression of the hairy-related basic helix-loop-helix gene HeyL in wild-type and Notch pathway mutant mice. We show that HeyL is strongly expressed in the presomitic mesoderm, the somites, the peripheral nervous system and smooth muscle of all arteries. Loss of HeyL expression at the level of nascent somites in Notch1 and Delta-like1 knockout mutants implicates HeyL as a Notch effector during somite formation. Furthermore, HeyL expression in vascular smooth muscle cells and in the thymus strikingly overlaps with that of Notch3, mutations of which underlie the CADASIL vascular disorder. PMID- 11044626 TI - MAEG, an EGF-repeat containing gene, is a new marker associated with dermatome specification and morphogenesis of its derivatives. AB - We report on the expression pattern of a novel EGF- containing gene named Maeg. RNA in situ studies indicate that Maeg is first activated during specification of the early lateral dermatome, and continues to be expressed in all the dermatome derivatives as the dermis of the trunk, the hair follicles, and the mesenchyme of the cranio-facial region. PMID- 11044627 TI - Expression of receptor tyrosine phosphatase gamma during early development of the chick embryo. AB - Studies in Drosophila suggest that receptor-tyrosine phosphatases are key regulators of neural development, however little is known about their expression or function in the nervous system of vertebrate embryos. In this report, we describe the expression pattern of RPTPgamma during early chick embryogenesis. Transcripts are largely restricted to the developing nervous system including oculomotor, trochlear and branchiomotor populations but are absent from spinal motor neurones. RPTPgamma is also detected in cells in the positions of hindbrain reticulospinal neurones, spinal commisural neurones and in cells with neuronal morphology in the ventral diencephalon. Within the peripheral nervous system transcripts are found in neuroblasts delaminating from epibranchial placodes and subsequently in placode-derived cranial ganglia. Outside the nervous system expression is detected in somites and transiently in the second branchial arch and the cranial mesenchyme. PMID- 11044628 TI - The expression of teneurin-4 in the avian embryo. AB - The teneurins are a family of four transmembrane proteins expressed in the developing vertebrate nervous system, though the Drosophila teneurin ten-m is also a pair-rule gene. Whole-mount in situ hybridization was used to localize teneurin-4 transcripts in the chicken embryo. The earliest signal is detected at stage 19 in the somites and limb buds. By stage 20 teneurin-4 transcripts are detected in temporal periocular mesenchyme, branchial arches, diencephalon and somites. Teneurin-4 expression in the limbs changes dramatically during development. Between stages 19 and 21 teneurin-4 expression is concentrated proximally in the zone of polarizing activity. Between stages 24 and 26 teneurin 4 is expressed in the mesenchyme of the anterodistal part of the limb. As in Drosophila, vertebrate teneurins are expressed not only in the nervous system, but also in non-neuronal tissues during pattern formation. PMID- 11044629 TI - Erratum to "Parathyroid hormone-related peptide is involved in protection against invasion of tooth germs by bone via promoting the differentiation of osteoclasts during tooth development" PMID- 11044630 TI - Corn husk oil lowers plasma LDL cholesterol concentrations by decreasing cholesterol absorption and altering hepatic cholesterol metabolism in guinea pigs. AB - To test the hypocholesterolemic mechanisms of corn husk oil (CoHO), male Hartley guinea pigs were fed diets containing increasing doses of CoHO, either 0 (control), 5, 10, or 15 g/100 g, and 0.25 g/100 g cholesterol. A positive control group (LC) with low dietary cholesterol (0.04 g/100 g) was also included. Fat was adjusted to 15 g/100 g in all diets by the addition of regular corn oil. Plasma low density lipoprotein (LDL) cholesterol concentrations were 32, 55, and 57% (P < 0.0005) lower with increasing doses of CoHO. In addition, intake of CoHO resulted in 32 to 43% lower hepatic total and esterified cholesterol and 55 to 60% lower triacylglycerol concentrations compared with the control group (P < 0.01). CoHO intake resulted in plasma and hepatic cholesterol concentrations similar to those in guinea pigs from the LC group. The number of cholesteryl ester and free cholesterol molecules was higher in LDL from the control group than in LDL from the CoHO or the LC groups. Hepatic beta-hydroxy-beta methylglutaryl-coenzyme A reductase activity was not modified by CoHO intake whereas cholesterol 7alpha-hydroxylase was up-regulated by 45 to 49% (P < 0.01) in the 10 and 15 g/100 g CoHO groups. Hepatic acyl coenzyme A cholesterol acyltransferase activity was down-regulated in a dose-dependent manner by 54, 58, and 63% with increasing doses of CoHO. CoHO intake resulted in increased fecal cholesterol excretion by 40 to 55% compared with the control and LC groups. Total fecal neutral sterol excretion was enhanced 42 to 55% by CoHO compared with the control group and by 59 to 68% compared with the LC group. The data from these studies suggest that CoHO has its hypocholesterolemic effect by decreasing cholesterol absorption and increasing bile acid output. These alterations in the intestinal lumen alter hepatic cholesterol metabolism and may affect the synthesis and catabolism of lipoproteins. PMID- 11044631 TI - Changes of transferrin-free iron uptake by bone marrow erythroblasts in strenuously exercised rats. AB - The effects of strenuous exercise on transferrin-free iron (Fe II) uptake by bone marrow erythroblasts in rats were investigated. Female Sprague-Dawley rats were randomly assigned to one of six groups, three of which underwent 3, 6, or 12 months of strenuous exercise (swimming 2 hr/day, 5 days/week) or their corresponding three control groups. At the end of experiments, bone marrow erythroblasts were isolated for Fe II uptake assay in vitro. It was found that the amounts of iron uptake into cytosole and stroma of the cells of rats in the groups undergoing 3 and 12 months of exercise did not differ from those in their corresponding sedentary groups. In addition, analysis of nonspecific and specific Fe II uptake by cytosole and stroma did not display any significant difference between the exercise and corresponding sedentary groups. However, the amount of Fe II uptake into cytosole and stroma was significantly increased in rats that exercised for 6 months compared with the corresponding controls. Nonspecific iron uptake into stroma was significantly higher in the exercise group than in the sedentary group (0.120 +/- 0.018 vs. 0.049 +/- 0.006 pM/10(6) cells, P < 0.01). The V(max) of the specific iron uptake into stroma was significantly higher (0.326 +/- 0.024 vs. 0.238 +/- 0.037 pM/10(6) cells, P < 0.05) and the K(m) of iron uptake into cytosol lower (0.08 +/- 0.01 vs. 0.21 +/- 0.03 microM, P < 0.001) in the exercise groups than in the control groups. These results indicate that 6 months of strenuous exercise could significantly increase Fe II uptake by the cells, probably by affecting the number and/or affinity of the putative iron carrier and the permeability to iron of cell membrane. The increased ability of cell-free iron accumulation in exercise might be a self-protective mechanism for body cells from the free iron-induced free radical reaction in addition to providing more iron for cell heme synthesis. PMID- 11044632 TI - Hepatic fructose-metabolizing enzymes and related metabolites: role of dietary copper and gender. AB - The purpose of this study was to further examine the hypothesis that variations in hepatic fructose-metabolizing enzymes between males and females might account for the differences in the severity of copper (Cu) deficiency observed in fructose-fed male rats. Weanling rats of both sexes were fed high-fructose diets either adequate or deficient in copper for 45 days. Cu deficiency decreased sorbitol dehydrogenase activity and dihydroxyacetone phosphate levels and increased glyceraldehyde levels in both sexes. Gender effects were expressed by higher activities of glycerol 3-phosphate dehydrogenase and aldehyde dehydrogenase in male than in female rats and higher levels of dihydroxyacetone phosphate and fructose 1,6-diphosphate (F1,6DP) in female than in male rats. The interactions between dietary Cu and gender were as follows: alcohol dehydrogenase activities were higher in female rats and were further increased by Cu deficiency in both sexes; aldehyde dehydrogenase activities were decreased by Cu deficiency only in male rats; sorbitol levels were higher in male rats and were further increased by Cu deficiency in male rats; fructose 1-phosphate (F1P) levels were increased by Cu deficiency in both sexes, but to a greater extent in male rats; glyceraldehyde 3-phosphate levels were higher in female rats, but were decreased by Cu deficiency in female and increased in male rats. Though most of the examined hepatic fructose-metabolizing enzymes and metabolites showed great differences between rats fed diets either adequate or deficient in Cu, it is the activity of fructokinase and aldolase-B, and the concentrations of their common metabolites, F1P and notably F1,6DP, that could be in part responsible for differences in the severity of pathologies associated with Cu deficiency observed between female and male rats. PMID- 11044633 TI - Effects of dietary n-3 fatty acids on contractility, Na+ and K+ currents in a rat cardiomyocyte model of arrhythmia. AB - The n-3 polyunsaturated fatty acids (PUFAs) have been reported to prevent ventricular fibrillation in human clinical studies and in studies involving experimental animals and isolated cardiomyocytes. This study aimed to determine whether dietary n-3 PUFAs could prevent isoproterenol and free radical-induced arrhythmic (asynchronous) contractile activity in adult rat cardiomyocytes and whether whole-cell Na(+) and K(+) currents measured by patch-clamp techniques were affected. Dietary supplementation with fish oil for 3 weeks significantly increased the proportion of total n-3 PUFAs in ventricular membrane phospholipids compared with saturated fat supplementation (18.8 +/- 0.6% vs. 8.1 +/- 1.0%, respectively). Cardiomyocytes from the fish oil group were less susceptible to isoproterenol-induced asynchronous contractile activity than were those from the saturated fat group [EC(50) values: 892 +/- 130 nM, n = 6 and 347 +/- 91 nM, n = 6 (P < 0.05), respectively]. Fish oil supplementation also prolonged the time taken to develop asynchronous contractile activity induced by superoxide and hydrogen peroxide. The voltage dependence of inactivation of Na(+) currents were significantly altered (-73.5 +/- 1.2 mV, n = 5 vs. -76.7 +/- 0.7 mV, n = 5, P < 0.05, for saturated fat and fish oil treated groups, respectively). The voltage dependence of activation of Na(+) and K(+) currents was not significantly affected by the dietary fish oil treatment. These results demonstrate the antiarrhythmic effects of dietary fish oil in a cardiomyocyte model of arrhythmia. PMID- 11044634 TI - Starvation alters the activity and mRNA level of glutaminase and glutamine synthetase in the rat intestine. AB - The metabolism of glutamine, the main respiratory fuel of enterocytes, is governed by the activity of glutaminase and glutamine synthetase. Because starvation induces intestinal atrophy, it might alter the rate of intestinal glutamine utilization. This study examined the effect of starvation on the activity, level of mRNA, and distribution of mRNA of glutaminase and glutamine synthetase in the rat intestine. Rats were randomized into groups and were either: (1) fed for 2 days with rat food ad libitum or (2) starved for 2 days. Standardized segments of jejunum and ileum were removed for the estimation of enzyme activity, level of mRNA, and in situ hybridization analysis. The jejunum of the fed rats had a greater activity of both enzymes per centimeter of intestine (P < 0.01), a greater glutaminase specific activity (1.97 +/- 0.45 vs. 1.09 +/- 0.34 micromol/hr/mg protein, P < 0.01), and a lower level of glutaminase and glutamine synthetase mRNA. The ileum of the fed rats had a greater activity of glutamine synthetase per centimeter of intestine (162.9 +/- 50.6 vs. 91.0 +/- 23.1 nmol/hr/cm bowel, P < 0.01), a lower level of glutaminase mRNA, and a greater level of glutamine synthetase mRNA. In situ hybridization analysis showed that starvation does not alter the distribution of glutaminase and glutamine synthetase mRNA in the intestinal mucosa. This study confirms that starvation decreases the total intestinal activity per centimeter of both glutaminase and glutamine synthetase. More importantly, the results indicate that the intestine adapts to starvation by accumulating glutaminase mRNA. This process prepares the intestine for a restoration of intake. PMID- 11044635 TI - Influence of one bout of vigorous exercise on ascorbic acid in plasma and oxidative damage to DNA in blood cells and muscle in untrained rats. AB - We investigated the influence of a single exhaustive bout of downhill running on oxidative damage to DNA and changes of antioxidant vitamin concentrations in rats. Plasma vitamin E levels were unchanged up to 48 hr postexercise. However, plasma ascorbic acid (AA) levels increased after the exercise, then decreased thereafter. This increase corresponded to a marked decrease in AA concentration in the adrenal glands. The activity of hepatic l-gulono-gamma-lactone oxidase, which catalyzes AA synthesis, was unaltered after the exercise. The weight of the adrenal glands was significantly increased 24 hr postexercise. These results indicate that the change in the plasma AA concentration after vigorous exercise was due mainly to the release of AA from the adrenal glands. The plasma creatine phosphokinase (CPK) activity and white blood cell (WBC) count increased 3 to 6 hr postexercise. Over this same period, a marker of oxidative DNA damage, 8 hydroxydeoxyguanosine in DNA, increased in the WBC, but not in the foreleg muscle. Lipid peroxide and vitamin E levels were also unchanged in the foreleg muscle. There was a positive correlation between CPK activity in the plasma and DNA damage in the WBC, suggesting that the DNA damage in the WBC was closely related with muscle damage due to exercise. PMID- 11044636 TI - A residential study comparing the effects of diets rich in stearic acid, oleic acid, and linoleic acid on fasting blood lipids, hemostatic variables and platelets in young healthy men. AB - Dietary fat is known to influence the variables of blood coagulation and fibrinolysis associated with vascular disease. However, the role of fat content and/or fat composition of the diet in this regard is still not well understood. In the present study, we investigated the effects of three isoenergic diets of differing fat composition in nine healthy young men in a strictly controlled residential study. Subjects consumed the three experimental diets for periods of 2 weeks each, separated by a washout period of at least 5 weeks in a randomized crossover design. The diets provided 38% of total energy intake as fat, 45% as carbohydrate, and 17% as protein, and differed only with respect to the fatty acid composition (stearic acid-rich diet: 34.1% stearic acid, 36.6% oleic acid; oleic acid-rich diet: 65.8% oleic acid; linoleic acid-rich diet: 36.5% linoleic acid, 38% oleic acid). Blood samples were collected at the beginning and at the end of each dietary period from fasted subjects for determination of factor VII coagulant activity (FVIIc), activated factor VII (FVIIa), factor VII antigen (FVIIag), tissue plasminogen activator (tPA) activity, plasminogen activator inhibitor type 1 (PAI-1) activity, fibrinogen, prothrombin fragment 1+2 (F(1+2)), and plasma lipids. There were no significant differences between diets in fasting plasma concentrations of FVIIc, FVIIa, FVIIag, fibrinogen, F(1+2), PAI-1 activity, and tPA activity. Plasma concentrations of lipids (high density lipoproteins, low density lipoproteins, triacylglycerols, and total cholesterol) were also unaffected. Although there were no changes in platelet aggregation response and membrane fluidity observed in any of the diets, increased anti aggregatory prostaglandin E(1) binding to platelet membranes was observed only in the case of linoleic acid-rich diet. In conclusion, diets with very different fatty acid compositions, at 38% of energy as fat intake, did not significantly influence blood coagulation, fibrinolysis, or blood lipids in the fasting state in young healthy men. PMID- 11044637 TI - The art and science of prognosis in patients with advanced cancer. PMID- 11044638 TI - What does positron emission tomography offer oncology? AB - The origins of positron emission tomography (PET) date back 70 years. Since the 1970s, however, its use has increased exponentially in the fields of neurology, cardiology and oncology. [18F]-Fluorodeoxyglucose (FDG) whole-body scanning is by far the most widely utilised and recognised application of PET in oncology. However, PET is a very versatile and powerful imaging modality capable of helping bridge the gap between the laboratory and the clinic. This article reviews the history and current applications of PET in oncology and then explores some of the newer applications and potential future uses of this versatile technology particularly in the area of cancer research. PMID- 11044639 TI - Terminal cancer. duration and prediction of survival time. AB - The duration of the terminal period of cancer allows us to determine its prevalence, which is necessary to plan palliative care services. Clinical prediction of survival influences access to palliative care and the healthcare approach to be adopted. The objective of this study was to determine the duration of the terminal period, the prognostic ability of healthcare professionals to predict this terminal period and the factors that can improve the prognostic accuracy. In the island of Mallorca, Spain, we followed 200 cancer patients at the inception of the terminal period. Twenty-one symptoms, quality of life, prognosis and duration of survival were measured. Using a Cox regression model, a predictive survival model was built. Median duration was 59 days; 95% confidence interval (CI)=49-69 days, mean=99 days. The oncologists were accurate in their predictions (+/-1/3 duration) in 25.7% of cases, the nurses in 21.5% of cases and the family physicians in 21.7% of cases. Errors of overestimation occurred 2.86 4.14 times more frequently than underestimation. In the final model, in addition to clinical prognosis (P=0.0094), asthenia (P=0.0257) and the Hebrew Rehabilitation Centre for Aged Quality of Life (HRCA-QL) Index (P=0.0002) were shown to be independent predictors of survival. In this study, the estimated duration of the terminal period was greater than that reported in a series of palliative care programmes, and survival was overestimated. Oncologists could estimate prognosis more accurately if they also take into account asthenia and HRCA-QL Index. PMID- 11044640 TI - A clinical nomogram for predicting long-term survival in advanced colorectal cancer. AB - From our prospectively accrued database of patients with gastrointestinal cancer, 1057 patients with advanced colorectal cancer were identified with the aim of determining predictive factors for survival of greater than 2 years and to use this information to develop a predictive nomogram. Patient's baseline characteristics, type and number of chemotherapy regimens received, and response to chemotherapy were assessed by univariate and multivariate logistic regression comparing those who survived greater than or less than 2 years. A total of 161 (15.2%) patients survived more than 2 years, so-called long survivors (LS). In multivariate analysis, positive predictive factors for LS were: good performance status (PS), normal serum carcinoembryonic antigen (CEA), rectal primary, Dukes' stage A-B, well or moderate differentiation, two or less disease sites, response to chemotherapy and treatment used protracted venous infusion (PVI) 5 fluorouracil (5-FU) in first-line chemotherapy, and the increasing number of chemotherapy treatments received. From these PS, CEA, number of sites and response to first-line chemotherapy were used to develop a nomogram capable of predicting the probability of survival beyond 2 years for an individual patient. This large study confirmed the relevance of known prognostic factors in metastatic colorectal cancer and demonstrated the importance of response to chemotherapy as an independent factor to predict LS. By combining these, we developed a nomogram which provides information which is likely to prove useful in the management of patients with advanced colorectal cancer. PMID- 11044641 TI - Prognostic significance of TP53 gene mutation in 995 cases of colorectal carcinoma. Influence of tumour site, stage, adjuvant chemotherapy and type of mutation. AB - Previous studies on the prognostic significance of TP53 gene alterations in colorectal cancer (CRC) have led to conflicting results. The present study investigated the prognostic significance of TP53 gene mutation in a very large series of 995 Dukes' B and C CRC patients, the majority of whom did not receive chemotherapy. Mutations were found in 385 (39%) cases and were not associated with tumour stage, histological grade, patient age or sex. Significantly more mutations were found in tumours from the left-sided colon compared with those from the right side (43% versus 34%, P=0.006). TP53 gene mutation had no prognostic value in the overall series or in different site or stage subgroups. None of the different types of TP53 gene mutation showed prognostic value. A trend for association with worse survival was observed in the patient subgroup that received adjuvant chemotherapy (Hazard Ratio (HR) 1.4, 95% confidence interval (CI) 0.89-2.21, P=0.15). These results indicate that mutation of the TP53 gene is not a useful prognostic marker for CRC patients who do not receive adjuvant chemotherapy. Further study is required to determine whether different types of TP53 mutation might be of value in predicting the response of CRC patients to chemotherapy. PMID- 11044642 TI - Ovarian cancer. an institutional review of patterns of care, health insurance and prognosis. AB - The purpose of this study was to investigate the prognostic importance of the health insurance status in 145 consecutive patients with ovarian cancer diagnosed between 1984 and 1996. All patients had basic (Type III) insurance to cover outpatient treatment and hospital expenses for a per diem flat fee; some patients had one of two types of supplemental private insurance (Type I and Type II) to cover the treatment by physicians of their choice and fee-for-service hospital treatment. The prognostic impact of health insurance was evaluated by multivariate statistical methods. The median follow-up was 81.9 months (range: 21 181); the 5-year probability of survival was 72% (standard error of the mean (SEM) 9.8%) for stage I, 53% (SEM 16.2%) for stage II, 17% (SEM 5. 9%) for stage III and 11% (SEM 5.5%) for stage IV cancer. Age, stage, histological grade and debulking surgery were independent predictors of survival in multivariate proportional hazards regression analysis. Patients with private insurance were younger and received more chemotherapy than patients with basic insurance. In multivariate analysis, insurance was an independent predictor of survival: patients with Type II insurance had a hazard ratio of 2.31 (95% confidence interval (CI): 1.05-5.04), and patients with Type III insurance had a hazard ratio of 3.30 (95% CI 1.52-7.17) compared with the reference group of Type I insured patients. Health insurance status was an independent predictor of survival in ovarian cancer. Research is needed to devise strategies to improve the medical care of patients with basic insurance. PMID- 11044643 TI - Prospective evaluation of the contribution of K-ras mutational analysis and CA 19.9 measurement to cytological diagnosis in patients with clinical suspicion of pancreatic cancer. AB - The aim of this study was to prospectively evaluate the diagnostic contribution of the detection of K-ras mutation and measurement of serum CA 19.9 concentrations to cytological diagnosis in patients with clinical suspicion of pancreatic cancer. These patients had either the presence or absence of a pancreatic mass as determined by imaging procedures. A total of 156 consecutive patients with clinical suspicion of pancreatic cancer or for confirmation and follow-up of their chronic pancreatitis disease were included: 84 patients presenting a pancreatic mass (group 1) and 72 patients without a pancreatic mass (group 2). K-ras mutations were detected by a restriction fragment length polymorphism/polymerase chain reaction (RFLP/PCR) method and CA 19.9 by an immunoluminometric assay. When a pancreatic mass was present, cytology offered a high sensitivity, but with a significant number of inconclusive results and K-ras mutational analysis offered a highly specific test. In the absence of a pancreatic mass, CA 19.9 (cut-off 100 U/ml) increased the sensitivity of the diagnosis by cytology and K-ras mutational analysis did not add significant information. Thus both tests contribute to the clinical decision process when pancreatic cancer is clinically suspected and the cytological report is not conclusive. PMID- 11044645 TI - BRCA1 mutations and clinicopathological features in a sample of Italian women with early-onset breast cancer. AB - Breast cancer in young women is uncommon and often presents with unfavourable biopathological features. Although early age at onset could suggest a genetic susceptibility to cancer, the appropriateness of BRCA1 testing for women with early-onset breast cancer and modest family history (FH) is controversial. 40 Women diagnosed with breast cancer at the age of 35 years or less, unselected for FH, were screened for germ line BRCA1 mutations by automated sequencing of exons 2, 5, 6, 11, 13 and 20. Overall, deleterious mutations were evidenced in 6 (15%) patients. With regard to FH, mutations were detected in 14%, 11% and 29% of women with none, weak and strong FH, respectively. Large tumour size, grade 3, lack of oestrogen receptors and high proliferation rate were significantly more common in mutation carriers (MC). Our data support both the appropriateness of testing young breast cancer patients and the frequency of unfavourable features in BRCA1 related breast cancer. It is hypothesised that BRCA1 mutations partially justify the high rate of aggressive breast cancer in young patients and that combining age and breast cancer phenotype could help to identify probable MC. PMID- 11044644 TI - Germ line BRCA1 and BRCA2 gene mutations in Turkish breast cancer patients. AB - Germ line BRCA1 and/or BRCA2 mutations were screened in 50 Turkish breast and/or ovarian cancer patients composed of hereditary, familial, early onset and male cancer groups. Genomic DNA samples were tested by heteroduplex analysis and DNA sequencing. Two truncating BRCA2 mutations, one novel (6880 insG) and one previously reported (3034 delAAAC), were found in two out of six (33%) hereditary breast and/or ovarian cancer patients. A novel truncating (1200 insA) and a missense (2080A-->G) BRCA1 mutation was found in two of 27 (7%) individuals in the early onset group. A total of four (8%) disease-causing mutations in 50 breast cancer patients were identified in BRCA1 and BRCA2 genes. In addition, five BRCA1 sequence variants have been identified in 23 patients. These results indicate that BRCA1 and BRCA2 genes are involved in some, but not all, forms of hereditary predisposition to breast cancer in the Turkish population. PMID- 11044646 TI - The attitudes of physicians and oncologists towards unconventional cancer therapies (UCT). AB - Physicians represent the main providers of unconventional cancer therapies (UCT) in Germany. However, little is known about providers' characteristics, as well as their attitudes towards UCT. 833 questionnaires on this topic answered by general practitioners and hospital physicians were analysed. Providers differed significantly from non-providers with respect to gender (male>female, i.e. more male providers), age (older>younger), amount of subjective knowledge about UCT, place of work (office>hospital>university clinic), greater wish for coverage of UCT costs, the belief in future positive trends concerning UCT, the recognition of patients' demand for UCT, the number of patients seen per month and medical specialty (GPs>oncologists and radiation oncologists). UCT were not considered to be highly effective, but estimations varied considerably. Further investigations in this area, better education about UCT, training in coping strategies with the fate of cancer patients, and reasonable complementary treatments appear to be of the utmost importance. PMID- 11044647 TI - Information and communication in the context of a clinical trial. AB - The aim of this study was to determine the communicative needs of the patients in the context of being invited to participate in a clinical trial. A questionnaire was sent to 299 patients with breast cancer randomised in a trial of adjuvant therapy. It was returned by 261 (87%) of them. Ninety-one per cent (231/255) of the patients regarded the information provided as easy or quite easy to understand. However, the method of treatment allocation was unclear to most patients: 51% (128/251) thought that the doctor had chosen the treatment while only 23% (57/251) knew that they had been randomised. Younger and better educated patients had a better understanding. For 55% (125/226) of the patients written information had been helpful in decision making. This correlated highly with the education of the patient. Sixty-eight per cent (174/255) of the patients thought that they had enough time for decision-making. Less educated patients and older patients had needed more time. Eighty-seven per cent (218/251) were happy with their decision to participate. While most patients are satisfied with the information received, there is a poor understanding of how treatment is allocated. Information should be modified for older and less-educated patients. The needs of the patients when offered participation in a clinical trial are clear information, enough time to consider the options and psychological support. PMID- 11044648 TI - Abundant expression of CD40 and CD40-ligand (CD154) in paediatric Langerhans cell histiocytosis lesions. AB - The pathogenesis of Langerhans cell histiocytosis (LCH) is obscure, partly because the events leading to activation of Langerhans-like lesional cells (LCH cells) and associated T cells, and the excessive cytokine production by these cells are unknown. The interaction between CD40 on antigen-presenting cells (APC) like Langerhans cells and CD40 ligand (CD40L) (CD154) expressed by activated CD4+ T cells, is essential for the activation of both the APC and the T cells and results in upregulation of APC functions and initiation of immunoreactivity. The effects of CD40-CD40L interaction include increased expression of co-stimulatory and adhesion molecules, proliferation, and production of pro-inflammatory cytokines and proteolytic enzymes, all features of LCH. Using immunohistochemistry, we analysed the in situ presence of the co-stimulatory molecules CD40 and CD40L in 15 fresh frozen biopsies of LCH lesions in children. The cells producing these molecules were identified by double staining for CD1a on LCH cells and CD3 on T cells. Prominent expression of CD40 by LCH cells and CD40L by T cells was found in all 15 specimens regardless of the source of specimen or characteristics of the patient. The findings of high expression of CD40 and CD40L in all specimens imply a key role for the CD40-CD40L adhesion pathway in the pathogenesis of LCH. Since this interaction is an accessible and realistic target for immunotherapy, these findings prompt speculation on the use of blocking antibodies to CD40 or to CD40L in the treatment of LCH. PMID- 11044649 TI - Family history of colorectal adenomatous polyps as a risk factor for colorectal cancer. AB - The aim of this study was to evaluate the risk of common colorectal cancer among first-degree relatives of patients with colorectal adenomatous polyps. In a population screening programme, 59406 subjects underwent an immunochemical faecal occult blood test. In a medical check-up-based cross-sectional study, 6139 subjects had a colonoscopic examination. They were divided into two groups, according to the results of a questionnaire on family history of colorectal adenomatous polyps, and the detection rates for colorectal cancer were compared in the groups positive or negative for a family history of colorectal adenomatous polyps. In the screening programme-based cross-sectional study, the detection rate for colorectal cancer was 0.57% (95% confidence interval (CI): 0.38-0.76) and 0.15% (95% CI: 0.12-0.18) in subjects with and without a family history of colorectal adenomatous polyps, respectively, showing a significant difference in the detection rate for colorectal cancer between the two groups (P<0.05). In the medical check-up-based cross-sectional study, the detection rate for colorectal cancer was 2.31% (95% CI: 1.15-3.47) and 0.53% (95% CI: 0. 34-0.72) in subjects with and without a family history of colorectal adenomatous polyps, respectively, indicating a significant difference between the two groups (P<0.05). These findings indicate that first-degree relatives of patients with colorectal adenomatous polyps have an elevated risk for common colorectal cancer, and that people with a family history of colorectal adenomatous polyps should be considered as a priority group for colorectal cancer screening. PMID- 11044650 TI - Selected micronutrients and colorectal cancer. a case-control study from the canton of Vaud, Switzerland. AB - The association between dietary intake of various micronutrients and colorectal cancer risk was analysed using data from a case-control study conducted between 1992 and 1997 in the Swiss Canton of Vaud. Cases were 223 subjects (142 (64%) males, 81 (36%) females; median age 63 years) with incident, histologically confirmed colon (n=119; 53%) or rectal (n=104; 47%) cancer, and controls were 491 subjects (211 (43%) males, 280 (57%) females; median age 58 years; range 27-74) admitted to the same university hospital for a wide spectrum of acute non neoplastic conditions, unrelated to long-term modifications of diet. Dietary habits were investigated using a validated food frequency questionnaire (FFQ). Odds ratios (OR) were obtained after allowance for age, sex, education, smoking, alcohol, body mass index, physical activity, and total energy and fibre intake. No significant association was observed for calcium, retinol, folate, vitamin D or E. The risk of colorectal cancer was directly associated with measures of iron intake (OR=2.43 for the highest tertile, 95% confidence interval (CI): 1.2-5.1) and inversely associated with vitamin C (OR=0.45; 95% CI: 0.3-0.8), and non significantly with total carotenoids (OR=0.66, 95% CI: 0.4-1.1). Among various individual carotenoids considered, inverse associations were observed for alpha carotene, beta-carotene and lutein/zeaxanthin. These findings were consistent across the strata of gender and age, and support the hypothesis that selected micronutrients have a favourable effect on colorectal carcinogenesis. PMID- 11044651 TI - Differential activity of topotecan, irinotecan and SN-38 in fresh human tumour cells but not in cell lines. AB - The topoisomerase I inhibitors topotecan irinotecan (CPT-11) and its metabolite SN-38 were studied in a panel of cell lines and in primary tumour cells from patients, using a non-clonogenic cytotoxicity assay. All three substances showed similar activity patterns in the panel of cell lines established to classify the drugs mechanistically. In the patient tumour cells the drugs had different effects. In haematological and ovarian cancer samples, SN-38 was much more potent than topotecan, followed by irinotecan, while in colorectal cancer samples only irinotecan showed substantial activity. This in vitro activity pattern seems to agree with clinical experiences to date. The inactivity of SN-38 in colorectal cancer suggests irinotecan may also have some other role in addition to being a prodrug to SN-38. This study raises questions as to the role and relevance of early preclinical model systems in anticancer drug development, and suggests that important information can be obtained from studies using primary cultures of human tumour cells. PMID- 11044652 TI - Increased upstream methylation has no influence on the overexpression of the parathyroid hormone-related protein gene in squamous cell carcinoma of the lung. AB - Humoral hypercalcaemia of malignancy (HHM) commonly results from the excessive production of a parathyroid hormone-related protein (PTHrP) by tumours. We have previously shown malignancy is associated with increased DNA methylation in the 5' region of the PTHrP gene. In a series of patients with lung carcinoma and relatively high serum calcium levels, 3 patients showed substantially increased PTHrP gene methylation while 5 patients showed no change in methylation status in this region. Patients showed marked tumour-specific expression of PTHrP through the P1 and P3 promoters with more general tumour and non-tumour expression through the P2 promoter. The lack of potential key regulatory CpG sites in the P1 promoter and the complete demethylation in the P2 and P3 promoters suggests methylation does not influence tumour-specific expression of PTHrP. Although demethylation may be a prerequisite for P2 and P3 expression, the overexpression of the PTHrP gene in cancer cells must be mediated through mechanisms other than DNA methylation. PMID- 11044653 TI - Attenuation of telomerase activity does not increase sensitivity of human melanoma cells to anticancer agents. AB - In tumour cells, replicative immortality is attained through stabilisation of telomeres by telomerase. Recent evidence suggests that telomerase plays an anti apoptotic role. Since apoptosis is the primary mode of cell death induced by several drugs, telomerase could be involved in determining the chemosensitivity profile of tumour cells. We investigated whether inhibition of telomerase activity through a hammerhead ribozyme targeting the RNA template of telomerase influences the susceptibility of human melanoma cells to a variety of anticancer agents (platinum compounds, taxanes, topoisomerase I inhibitors). The ribozyme sequence was inserted into an expression vector and the JR8 human melanoma cell line was transfected with it. The cell clones obtained showed a reduced telomerase activity. Growth inhibition curves generated after exposure of ribozyme-transfectant clones to individual drugs were superimposable to those obtained from parental cells. Moreover, telomerase inhibition did not promote apoptosis as a cellular response to drug treatment. Overall, our results indicate that downregulation of telomerase activity does not increase the sensitivity of melanoma cells to anticancer drugs. PMID- 11044654 TI - Selection of cancer chemopreventive agents based on inhibition of topoisomerase II activity. AB - The present study was undertaken to determine if in vitro inhibition of one or both of the two most dominant mammalian DNA topoisomerases (topos) is common among chemopreventive agents. To determine if an agent was a topo I inhibitor, we employed the DNA relaxation and nicking assays. For potential topo II inhibitors, we used the DNA unknotting and linearisation assays. 14 of 30 agents (47%) were ineffective in all four assays (IC(50) >100 microgram/ml), and 11 (37%) inhibited topo II catalytic activity. The sensitivity of the topo II assay was 63%, selectivity 93%, positive predictive value 91%, and total accuracy 77%. For chemopreventive efficacy, the positive predictive value of the unknotting assay was 92%, and the total accuracy was 60%. These data suggest that reduced topo II activity is a desirable property of many known chemopreventive agents. We conclude that the unknotting assay could be a valuable addition to the in vitro tests presently used to select chemopreventive agents. PMID- 11044655 TI - Cell cycle-related expression and ligand binding of peripheral benzodiazepine receptor in human breast cancer cell lines. AB - The aim of this study was to measure the expression of peripheral benzodiazepine receptors (PBR) as well as mitochondria content in different phases of the cell cycle of BT-20 and MCF-7 breast cancer cell lines, using two-parameter flow cytometric analyses. The PBR expression as well as mitochondria mass, were found to increase as cells pass through different stages of the cell cycle, whereas the amount of PBR in quiescent cells was very low. Binding capacity for the PBR ligand [3H]-Ro5-4864 was strongly related to the phase of the cell cycle with a positive correlation (r=0.98) with a high percentage of cells in S phase. Incubation of BT-20 cells in serum-deprived medium with nanomolar concentrations of Ro5-4864 caused an increase in S phase cells. This effect was not observed in MCF-7 cells. Using micromolar concentrations of Ro5-4864, both BT-20 and MCF-7 cells were reversibly arrested in the G(0/1) phase. PMID- 11044656 TI - Gelatinolytic activity of matrix metalloproteinase-2 and -9 in oesophageal carcinoma; a study using in situ zymography. AB - In this study, we investigated the activity of matrix metalloproteinase (MMP)-2 and -9 by gelatine zymography, immunostaining and in situ gelatine zymography in 30 oesophageal squamous-cell carcinomas. The gelatinolytic activity in situ was detected in all cases with different patterns of localisation. Significant gelatinolysis by stromal cells adjacent to tumour nests was found in 12 cases. Strong gelatinolytic activity appeared within the tumour nest itself in 13 cases. In the other 5 cases, both stromal cells and tumour cells showed the gelatinolytic activity. Gelatine zymography demonstrated a correlation between vascular invasion and activation of MMP-9. It also demonstrated a correlation between lymph node metastasis, lymphatic or vascular invasion and activation of MMP-2. These results suggest that MMPs play an important role in the invasion of oesophageal carcinoma. PMID- 11044657 TI - Immunohistochemical analysis of sporadic and familial (185delAG carriers) ovarian cancer in israel PMID- 11044658 TI - Erratum to 'Imaging of cancer invasion and metastasis using green fluorescent protein' eur J cancer 2000, 36(13), 1671-1680 PMID- 11044659 TI - Nitric oxide signalling in insects. PMID- 11044660 TI - Purification and properties of a beta-glycosidase purified from midgut cells of Spodoptera frugiperda (Lepidoptera) larvae. AB - Two beta-glycosidases (BG) (Mr 47,000 and Mr 50,000) were purified from Spodoptera frugiperda (Lepidoptera: Noctuidae) midguts. These two polypeptides associate or dissociate depending on the medium ionic strength. The Mr 47,000 BG probably has two active sites. One of the putative active sites (cellobiase site) hydrolyses p-nitrophenyl beta-D-glucoside (NPbetaGlu) (79% of the total activity in saturated enzyme), cellobiose, amygdalin and probably also cellotriose, cellotetraose and cellopentaose. The cellobiase site has four subsites for glucose residue binding, as can be deduced from cellodextrin cleavage data. The enzymatic activity in this site is abolished after carbodiimide modification at pH 6.0. Since the inactivation is reduced in the presence of cellobiose, the results suggest the presence of a carboxylate as a catalytic group. The other active site of Mr 47,000 BG (galactosidase site) hydrolyses p-nitrophenyl beta-D galactoside (NPbetaGal) better than NPbetaGlu, cleaves glucosylceramide and lactose and is unable to act on cellobiose, cellodextrins and amygdalin. This active site is not modified by carbodiimide at pH 6.0. The Mr 47,000 BG N terminal sequence has high identity to plant beta-glycosidases and to mammalian lactase-phlorizin hydrolase, and contains the QIEGA motif, characteristic of the family of glycosyl hydrolases. The putative physiological role of this enzyme is the digestion of glycolipids (galactosidase site) and di- and oligosaccharides (cellobiase site) derived from hemicelluloses, thus resembling mammalian lactase phlorizin hydrolase. PMID- 11044661 TI - Characterization of a leucokinin binding protein in Aedes aegypti (Diptera: Culicidae) Malpighian tubule. AB - The insect myokinin (leucokinin-like) neuropeptide family includes peptides that have different physiological effects such as the induction of hindgut myotropic activity and stimulation of urine production. The C-terminal pentamer of myokinins Phe-X-(Ser/Pro/Ala)-Trp-Gly-amide [X=Phe, His, Asn, Ser or Tyr], had been previously determined as the minimum fragment able to elicit a functional response. The receptor(s) for these insect neuropeptides has not yet been identified. In order to characterize the Malpighian tubule leucokinin-like peptide receptor(s) from the yellow fever mosquito (Aedes aegypti), a leucokinin photoaffinity analogue (LPA) of sequence dAla-dTyr-Bpa-dLys-Phe-Phe-Ser-Trp-Gly amide was designed based on structure/activity relationships for leucokinins. LPA caused depolarization of the transepithelial voltage (TEV) in female Malpighian tubule, confirming the activity of the peptide. The effective concentration to give half the maximum depolarization (EC(50)) was 17 nM. The (125)I-LPA was then used to characterize leucokinin binding proteins in female Malpighian tubule membranes. It specifically labeled and saturated a protein(s) of about 54 kDa as shown by SDS-PAGE/autoradiography and by competition experiments with excess unlabeled leucokinin analogues. (125)I-LPA bound to the 54 kDa protein(s) with a K(d) value of 13+/-3 nM in agreement with the EC(50) for the TEV bioassay. Altogether these data suggest that the 54 kDa protein is an Aedes-leucokinin receptor. This is the first characterization of an insect leucokinin receptor and reveals that LPA is a powerful tool to label insect myokinin receptors. PMID- 11044662 TI - Lipophorin as a yolk protein precursor in the mosquito, Aedes aegypti. AB - We examined expression of the lipophorin (Lp) gene, lipophorin (Lp) synthesis and secretion in the mosquito fat body, as well as dynamic changes in levels of this lipoprotein in the hemolymph and ovaries, during the first vitellogenic cycle of females of the yellow fever mosquito, Aedes aegypti. Lipophorin was purified by potassium bromide (KBr) density gradient ultracentrifugation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Polyclonal antibodies were produced against individual Lp apoproteins, apolipoprotein-I (apoLp-I) and apolipoprotein-II (apoLp-II), with molecular weights of 240 and 75 kDa, respectively. We report here that in the mosquito A. aegypti, Lp was synthesized by the fat body, with a low level of the Lp gene expression and protein synthesis being maintained in pre- and postvitellogenic females. Following a blood meal, the Lp gene expression and protein synthesis were significantly upregulated. Our findings showed that the fat body levels of Lp mRNA and the rate of Lp secretion by this tissue reached their maximum at 18 h post-blood meal (PMB). 20 Hydroxyecdysone was responsible for an increase in the Lp gene expression and Lp protein synthesis in the mosquito fat body. Finally, the immunocytochemical localization of Lp showed that in vitellogenic female mosquitoes, this protein was accumulated by developing oocytes where it was deposited in yolk granules. PMID- 11044663 TI - Metabolic pathways for diacylglycerol biosynthesis and release in the midgut of larval Manduca sexta. AB - The pathway for the synthesis of diacylglycerol in larval Manduca sexta midgut was studied. Fifth instar larvae were fed with [9, 10-(3)H]-oleic acid-labeled triolein and the incorporation of the label into lipid intermediates was analyzed as a function of time. The results showed that the triacylglycerol was hydrolyzed to fatty acids and glycerol in the midgut lumen. In midgut tissue, the labeled fatty acids were rapidly incorporated into phosphatidic acid, diacylglycerol and triacylglycerol, but no significant labeling of monoacylglycerol was observed. Dual-labeling experiments were performed in order to characterize the kinetics of diacylglycerol biosynthesis in the midgut, its incorporation into hemolymph lipophorin and its clearance from hemolymph. The results were best described by a model in which the rate-limiting step in diacylglycerol biosynthesis was the uptake of fatty acid from the lumen of the midgut. Once in the cell the fatty acid was rapidly incorporated in phosphatidic acid and diacylglycerol. Diacylglycerol was converted to triacylglycerol or exported into hemolymph. The interconversion of diacylglycerol and triacylglycerol was fairly rapid, suggesting that triacylglycerol serves as a reservoir from which diacylglycerol can be produced. This mechanism permits the cell to maintain a low steady-state concentration of diacylglycerol and yet efficiently absorb fatty acids from the lumen of the midgut. PMID- 11044664 TI - Inhibition of cysteine and aspartyl proteinases in the alfalfa weevil midgut with biochemical and plant-derived proteinase inhibitors. AB - Proteolytic activities in alfalfa weevil (Hypera postica) larval midguts have been characterized. Effects of pH, thiol activators, low-molecular weight inhibitors, and proteinase inhibitors (PIs) on general substrate hydrolysis by midgut extracts were determined. Hemoglobinolytic activity was highest in the acidic to mildly acidic pH range, but was maximal at pH 3.5. Addition of thiol activators dithiothreitol (DTT), 2-mercaptoethanol (2-ME), or L-cysteine had little effect on hemoglobin hydrolysis at pH 3.5, but enhanced azocaseinolytic activity two to three-fold at pH 5.0. The broad cysteine PI E-64 reduced azocaseinolytic activity by 64% or 42% at pH 5 in the presence or absence of 5 mM L-cysteine, respectively. Inhibition by diazomethyl ketones, Z-Phe-Phe-CHN(2) and Z-Phe-Ala-CHN(2), suggest that cathepsins L and B are present and comprise approximately 70% and 30% of the cysteine proteolytic activity, respectively. An aspartyl proteinase component was identified using pepstatin A, which inhibited 32% (pH 3.5, hemoglobin) and 50% (pH 5, azocasein) of total proteolytic activity. This activity was completely inhibited by an aspartyl proteinase inhibitor from potato (API), and is consistent with the action of a cathepsin D-like enzyme. Hence, genes encoding PIs with specificity toward cathepsins L, B and D could potentially be effective for control of alfalfa weevil using transgenic plants. PMID- 11044665 TI - Chitin is only a minor component of the peritrophic matrix from larvae of Lucilia cuprina. AB - The gut of most insects is lined with a peritrophic matrix that facilitates the digestive process and protects insects from invasion by micro-organisms and parasites. It is widely accepted that the matrix is composed of chitin, proteins and proteoglycans. Here we critically re-examine the chitin content of the typical type 2 peritrophic matrix from the larvae of the fly Lucilia cuprina using a range of techniques. Many of the histochemical and biochemical techniques indicate the presence of chitin, although they are often adversely influenced by the presence of highly glycosylated proteins, a principal component of the matrix. The alkali-stable fraction, which is used as an indicator of the maximum chitin content in a biological sample, is only 7.2% of the weight of the matrix. Larvae fed on the potent chitin synthase inhibitor polyoxin D or the chitin binding agent Calcofluor White, showed strong concentration-dependent inhibition of larval weight and survival but no discernible effects on the matrix structure. A bacterial endochitinase fed to larvae had no effect on larval growth and no observable effect in vitro on the structure of isolated peritrophic matrix. RT PCR did not detect a chitin synthase mRNA in cardia, the tissue from which PM originates. It is concluded that chitin is a minor structural component of the type 2 peritrophic matrix of this insect. PMID- 11044666 TI - Isolation and endocrine regulation of an HMG-CoA synthase cDNA from the male Jeffrey pine beetle, Dendroctonus jeffreyi (Coleoptera: Scolytidae). AB - We have isolated a full length 3-hydroxy-3-methylglutaryl coenzyme A synthase (HMG-S) cDNA from the male Jeffrey pine beetle, Dendroctonus jeffreyi Hopkins, and studied the effects of topical applications of juvenile hormone III (JH III) on its expression. The predicted translation product of this apparently single copy gene has 63% and 58% identity with HMG-S1 and HMG-S2 from Blattella germanica (L.), and 61% identity with Drosophila melanogaster Hmgs. HMG-S transcript levels remain uniformly low in JH III-treated and control D. jeffreyi females, but are induced approximately 2.5- to 5-fold in JH III-treated males. JH III causes a dose- and time-dependent increase in HMG-S transcripts in the male metathoracic-abdominal region. Since monoterpenoid pheromone precursor synthesis and HMG-CoA reductase expression are under the control of JH III in the metathorax of Ips bark beetles, the observed HMG-S expression pattern suggests that the isoprenoid pathway is similarly important for semiochemical production in D. jeffreyi. PMID- 11044667 TI - Cloning and characterization of a chitin synthase cDNA from the mosquito Aedes aegypti. AB - Characterization of the enzymes involved in the chitin biosynthetic pathway in mosquitoes is critical due to the importance of chitin in the formation of the peritrophic matrix [PM] and its potential impact on vector competence. Chitin is the homopolymer of the amino sugar N-acetyl-D glucosamine [GlcNAc]. The final step of incorporation of GlcNAc into the chitin polymer is catalyzed by the enzyme chitin synthase [CS]. CS is a membrane bound enzyme, but the mechanism of its action in the biosynthesis of the PM is not understood. We have isolated and sequenced a CS-encoding cDNA clone from the mosquito Aedes aegypti, compared its sequence with CS from other organisms and studied its RNA expression. The cDNA is 3.5 kb in length with an open reading frame of 2.6 kb that encodes a protein of 865 amino acids with a predicted molecular mass of 99.5 kDa. The putative translation product shares 90% similarity to two CS proteins from Caenorhabditis elegans and 50% similarity to Saccharomyces cerevisiae in the catalytic domain of CS enzymes. Data suggest that CS is a single copy gene. RT-PCR analysis shows CS message in whole non-blood-fed females, whole blood-fed females, non-blood-fed midguts and in midguts dissected at different time points post-blood-feeding. In situ hybridization studies of midgut samples revealed that CS mRNA increases following a bloodmeal and is localized to the periphery of the epithelial cells facing the midgut lumen. PMID- 11044668 TI - "Identification of a point mutation in an esterase gene in different populations of the southern cattle tick, boophilus microplus" by R. Hernandez, H. He, A. C. Chen, S. D. Waghela, G. W. Ivie, J. E. George and G. G. Wagner. Insect biochemistry and molecular biology 30(10) pp.969-977 PMID- 11044669 TI - The generation of diversity by Haemophilus influenzae. PMID- 11044670 TI - Virus receptor arrays, CD46 and human herpesvirus 6. PMID- 11044671 TI - The mycobacterial antigens 85 complex - from structure to function and beyond. PMID- 11044672 TI - Crowd control: quorum sensing in pathogenic E. coli. PMID- 11044673 TI - Gram-negative pathogens and molecular mimicry: is there a case for mistaken identity? PMID- 11044674 TI - NKT cells protect against pre-erythocytic malaria PMID- 11044675 TI - Bacterial cave dwellers PMID- 11044676 TI - Parallel evolution of pathogenic strains PMID- 11044677 TI - Sticking to mycobacteria PMID- 11044678 TI - Microbial genomics PMID- 11044679 TI - The Toll receptor family and microbial recognition. AB - The survival of multicellular organisms is dependent on their ability to recognize invading microbial pathogens and to induce a variety of defense reactions. Recent evidence suggests that an evolutionarily ancient family of Toll like receptors plays a crucial role in the detection of microbial infection and the induction of immune and inflammatory responses. PMID- 11044680 TI - Bacterial interplay at intestinal mucosal surfaces: implications for vaccine development. AB - The discovery of 'molecular syringes' in several important gastrointestinal pathogens including Escherichia coli, Salmonella, Shigella and Yersinia, together with a better understanding of M cells and the mucosal immune system, has advanced our appreciation of multistage microorganism-host cell interactions. Recent studies suggest that these molecular strategies could be adapted for the development of modular mucosal vaccines. PMID- 11044682 TI - Author correction PMID- 11044681 TI - Mechanisms of viral transport in the cytoplasm. AB - Analogous to the spread of viruses within the host animal during pathogenesis, from their site of entry to distant sites via the bloodstream, lymphatic system and nervous system, there is also movement within infected cells. As cytoplasmic diffusion only operates within very small volumes, active membrane traffic and cytosolic transport of viral genome-protein complexes are required, which involve both the actin and microtubule cytoskeleton. PMID- 11044683 TI - Immune-defense molecules of molluscum contagiosum virus, a human poxvirus. AB - Molluscum contagiosum virus encodes more than 150 proteins including some involved in host interactions that might contribute to prolonged viral replication in the skin. These include homologs of a selenocysteine-containing glutathione peroxidase, a death effector domain protein, a chemokine, a major histocompatibility complex class I molecule and an interleukin-18-binding protein. PMID- 11044684 TI - Growth of Legionella pneumophila in Dictyostelium discoideum: a novel system for genetic analysis of host-pathogen interactions. AB - Legionella pneumophila, the Gram-negative bacterium that causes Legionnaires' disease, can be cultured in the laboratory in a variety of fresh-water amoebae and macrophage-like cell lines. None of these hosts, however, is amenable to genetic analysis, which has limited the ability of researchers to analyse the host factors essential for L. pneumophila growth. In this article, we describe a novel method in which L. pneumophila is grown within the soil amoeba Dictyostelium discoideum and how D. discoideum genetics is being used to analyse the host cell factors involved in L. pneumophila pathogenesis. PMID- 11044685 TI - The Registry of the International Society for Heart and Lung Transplantation: seventeenth official report-2000. PMID- 11044686 TI - Treatment-seeking delays in heart failure patients. AB - BACKGROUND: Patients having cardiac symptoms often delay for hours before seeking treatment. Delay time is usually defined as the amount of time between the patient's first awareness of symptoms and arrival at the hospital. Excessive delays in seeking medical care for heart failure (HF) symptoms may influence patient outcomes. However, the treatment-seeking patterns of HF patients are not well understood. METHODS: We obtained data through a retrospective chart audit to describe the treatment-seeking behaviors of 753 HF patients, at a Veterans Administration facility, and to identify predictors of delay in seeking medical care for HF symptoms. Using univariate and multivariate analyses, we assessed relationships among delay time, presenting symptoms, and patient characteristics. RESULTS: The mean delay time was 2.93 +/- 0.68 days. The most common symptoms on admission were dyspnea (76%), edema (66%), fatigue (37%), and angina (25%). Variables negatively affecting delay time included presence of dyspnea and edema (odds ratio [OR], 2.10 and 1.82; confidence interval [CI], 1.38 to 3.19 and 1.17 to 2.82, respectively), care by a primary care physician (OR, 2.04; CI, 1.45 to 2.88), and higher New York Heart Association (NYHA) Class (OR, 1.96; CI, 1.47 to 2.61). Variables positively affecting delay time were the presence of chest pain (OR, 0.42; CI, 0.29 to 0.62) and a history of previous admission for HF (OR, 0.42; CI, 0.28 to 0.62). CONCLUSIONS: Delays in seeking treatment for HF symptoms are significantly high. This study supports the need for interventions that will increase early symptom recognition and management on the part of patients and their families. PMID- 11044687 TI - Response of right ventricular function to prostaglandin E1 infusion predicts outcome for severe chronic heart failure patients awaiting urgent transplantation. AB - BACKGROUND: Selection of patients for urgent heart transplantation who have end stage heart failure requires objective criteria to distinguish between subjects who may deteriorate clinically and those who can be stabilized. This population appears to differ in terms of right ventricular function, and right ventricular changes in loading may provide prognostic information. To investigate predictive parameters of patients admitted for urgent heart transplantation, we prospectively studied the mechanical performance of the right ventricle during acute afterload reduction. PATIENTS AND METHODS: We studied 68 heart failure patients hospitalized for bridge-to-transplant. The patients underwent right heart catherization at baseline and during prostaglandin E1 infusion. We stratified patients according to clinical outcome: Group A comprised patients who could be weaned from bridging therapy or who were electively transplanted after at least 90 days, and Group B comprised patients who died or who remained unstable and required transplant under urgent conditions. RESULTS: Right ventricular hemodynamics at baseline were comparable in both groups. However, during maximal vasodilatation, with prostaglandin E1 infusion, the right ventricular end-diastolic volume index (EDVI) was significantly reduced in Group A, (-23 ml/m(2) (p < 0.01) vs +12 ml/m(2) (p = n.s. DeltaEDVI in Group B. Reduction of EDVI by less than 10% during prostaglandin E1 infusion has a sensitivity of 89% and a specificity of 70% for clinical deterioration. CONCLUSIONS: The response of right ventricular volumes to prostaglandin E1 may predict the outcome of patients with severe congestive heart failure listed for urgent heart transplantation. PMID- 11044688 TI - Vagal reinnervation in the long term after orthotopic heart transplantation. AB - BACKGROUND: Sympathetic reinnervation after orthotopic heart transplantation (HTx) has become an accepted phenomenon, particularly in long-term transplanted patients. Parasympathetic reinnervation, however, still remains questionable. METHODS: In 38 HTx recipients, mean age of 51.6 +/- 9.7 years (range, 29 to 70 years), with a time span after HTx of 4.6 +/- 2.8 years (0.5 to 10.5 years), we stimulated carotid baroreceptors using periodic neck suction at low (0.1 Hz) and high (0.2 Hz) frequencies to test sympathetic and vagal responses, respectively. Respiratory rate was fixed at 0.25 Hz. We simultaneously recorded surface electrocardiogram, finger blood pressure, respiration and neck pressure signals while patients rested in the supine position. Time series of RR intervals, respiration, and neck and blood pressures were generated and subjected to spectral analysis. RESULTS: All patients demonstrated a 0.25-Hz peak in RR interval spectrum, caused by non-autonomic respiratory sinus arrhythmia. Thirteen patients (5. 2 +/- 3.5 years after HTx; range, 0.9 to 10.2 years) showed a baroreflex-induced sharp peak at 0.1 Hz in RR-interval power spectrum during 0.1 Hz neck suction, indicating sympathetic reinnervation. However at 0.2-Hz neck suction, 4 of the 13 sympathetically reinnervated patients displayed a baroreflex induced 0.2-Hz peak, which could be suppressed with atropine administration strong evidence for vagal reinnervation. CONCLUSIONS: Non-invasive carotid baroreflex stimulation is an appropriate tool to prove restoration of autonomic control after orthotopic HTx. Sympathetic reinnervation parallels parasympathetic reinnervation in long-term transplanted patients. PMID- 11044689 TI - Post-cardiac transplantation gout: incidence of therapeutic complications. AB - OBJECTIVE: To study the clinical impact of gout treatment following cardiac transplantation. METHODS: We performed an audit of all cardiac transplant recipients of the Alfred Hospital before August 1998 who lived in Victoria. RESULTS: We studied 225 patients (81% men), with a mean post-transplant follow-up of 50.8 months (SD 36). Forty-three (19%) had pre-transplant gout, 19 recurring post-transplantation. Twenty-three patients developed gout de novo. Of the 24 patients who received allopurinol, 6 developed pancytopenia and required hospitalization. Fourteen received a change in immunosuppression: in 5 patients following pancytopenia, and in 9 to enable safe use of allopurinol. Thirty-two patients received colchicine; 5 developed neuromyopathy. Impaired renal function, diuretic use, and hypertension were more common in this sub-group. Non-steroidal anti-inflammatory agents, used in 16 patients, caused serious complications in 1 patient (life-threatening peptic ulceration and hemorrhage, precipitating dialysis-dependent chronic renal failure). CONCLUSIONS: Cardiac transplant recipients, when treated for gout, are at high risk of therapeutic complications. Thus, gout treatment significantly affects care, health, and immunosuppression of these patients. PMID- 11044690 TI - Medium-term results of heart transplantation using older donor organs. AB - BACKGROUND: Donor heart shortage has necessitated the expansion of the donor pool by the use of older hearts. PATIENTS AND METHODS: In a 13-year period, 1,070 heart transplants were performed in 1,035 adults at the German Heart Institute Berlin. We divided the patients into 3 groups: Group I, donor age <35 years (n = 524); Group II, donor age 35 to 50 years (n = 379); Group III, donor age >50 years (n = 167). We analyzed post-operative mortality (up to 30 days), cumulative survival rates, cardiac dependent morbidity, and changes in the left/right ventricular ejection fraction as well as freedom from cytomegalovirus infection and freedom from acute rejection episodes grade >/= 2 (International Society for Heart and Lung Transplantation). We also calculated the rate of cardiac interventions per patient in the groups. RESULTS: Recipients in Group III were significantly older, compared with Groups I and II. The post-operative mortality was 16.8% in Group I, 29.8% in Group II, and 23.4% in Group III. The differences were significant (p = 0. 00001) between Group I and Group II. The long-term cumulative survival rates were significantly better in Group I when compared with Groups II and III (p < 0.00001, p = 0.014), but it did not differ between Groups II and III (p = 0.18). However, cardiac morbidity in Groups I and II was significantly lower when compared with Group III (p = 0.0009, p = 0.037). Mean left and right ventricular ejection fraction was >55% and did not significantly change in groups for up to 10 years. Freedom from cytomegalovirus infection was not significantly different between Groups II and III (p = 0.09). Significantly fewer percutaneous transluminal coronary angioplasties were performed in Group I, but comparable numbers were carried out in Groups II and III (p = 0.53). For retransplantation a similar situation occurred. CONCLUSION: We did not find significant differences in the mid-term follow-up between patients who received hearts from 35- to 50-year-old donors and from those who had received hearts from donors >50 years, despite increased cardiac morbidity in Group III. Close monitoring of the coronary situation after heart transplantation and expanded indications for revascularization in Group III makes heart transplantation with older hearts a suitable option to save the lives of patients in end-stage heart failure. PMID- 11044691 TI - Predictors of survival in patients bridged to transplantation with the thoratec VAD device: a single-center retrospective study on more than 100 patients. AB - BACKGROUND: Careful patient selection markedly influences the outcome of patients who undergo mechanical circulatory support. Therefore, we tried to evaluate predictors of survival after implantation of the Thoratec ventricular assist device (VAD). METHODS: Between October 1992 and January 2000, 104 patients (86 men, 18 women, aged 11 to 69 years) received the Thoratec VAD as a bridge to transplant. A total of 51 patients required left ventricular support (LVAD), 50 patients required biventricular support (BVAD), and 3 patients required total artificial heart implantation. We performed univariate analysis of 25 parameters with regard to their effect on survival and then applied a multivariate analysis to evaluate those factors that turned out to be marginally significant. We performed all analysis for the total collective as well as for the LVAD and BVAD sub-group. RESULTS: The BVAD patients tended to have worse outcomes than did LVAD patients. We found no significant predictors of survival in either sub-group. In the total collective, however, we found the following pre-implant conditions were independent risk factors for survival after VAD implantation: patient age > 60 years (odds ratio [OR] 3.87, confidence interval [CI] 1.39 to 10.76), pre-implant ventilation (OR, 6.76; CI 2.42 to 18.84), and increased pre-implant total bilirubin (OR, 1.42; CL, 1.19 to 1.69). CONCLUSIONS: Transplant candidates on inotropic support should be considered for bridging to transplant as soon as bilirubin values start to increase or before respiratory function deteriorates and ventilation becomes necessary. In elderly patients, careful patient selection, particularly considering potential risk factors, might favorably affect their outcomes. PMID- 11044692 TI - In the lung aerosol cyclosporine provides a regional concentration advantage over intramuscular cyclosporine. AB - BACKGROUND: Acute rejection remains an almost universal complication among lung transplant recipients. Refractory rejection as well as chronic systemic immunosuppression is associated with significant morbidity and mortality. Recent studies suggest that aerosol cyclosporine may address these issues by effectively preventing acute cellular rejection while maintaining low systemic drug concentrations. This study was designed to evaluate the concentrations of cyclosporine in blood and lung tissue after aerosol and intramuscular administration. METHODS: Lewis rats were divided into 4 experimental groups: Groups A (n = 33) and B (n = 30) received aerosol cyclosporine 3 and 5 mg/kg, respectively; Groups C (n = 33) and D (n = 30) received systemic cyclosporine 5 and 15 mg/kg, respectively. We used high-performance liquid chromatography to quantitate blood and lung tissue cyclosporine levels at timed intervals. We used the trapezoidal rule to approximate area under the concentration vs time curve (AUC). RESULTS: Aerosol delivery of cyclosporine resulted in higher and more rapid peak drug levels in lung tissue samples than did systemic delivery. At an equivalent 5 mg/kg dose, the cyclosporine AUC was 3 times higher with aerosol delivery than with intramuscular delivery in lung tissue (477,965 vs 157,706 ng x hour/g, respectively). The lung tissue: blood AUC ratio was highest in the aerosol groups (27.3:1 and 17.4:1) compared with the intramuscular groups (8.1:1 and 9.4:1). CONCLUSION: Local aerosol inhalation delivery of cyclosporine provides a regional advantage over systemic intramuscular therapy by providing higher peak concentrations and greater lung tissue exposure. PMID- 11044693 TI - Retrograde flush perfusion with low-potassium solutions for improvement of experimental pulmonary preservation. AB - BACKGROUND: Optimal preservation of post-ischemic organ function is a continuing challenge in clinical lung transplantation. Retrograde instillation of preservation solutions has the theoretic advantage of achieving homogeneous distribution in the lung because of perfusing both the pulmonary and the bronchial circulation. So far, we have seen no experimental studies that include stereologic analysis of intrapulmonary edema concerning the influence of retrograde preservation on post-ischemic lung function after preservation with Perfadex and Celsior. METHODS: In an extracorporeal rat model, we perfused 8 lungs, each, using either antegrade or retrograde perfusion technique with Celsior (CE(ant)/CE(ret)) and Perfadex (PER(ant)/PER(ret)). Results were compared with low-potassium Euro-Collins. Post-ischemic lungs were reventilated and reperfused mechanically. We continuously monitored relative oxygenation capacity (ROC), pulmonary artery pressure, flush time, and wet/dry ratio. Furthermore, we used stereologic means to evaluate edema formation. Statistics comprised different analysis of variance models. RESULTS: Relative oxygen capacity of CE(ant)-protected lungs was superior to that of PER(ant) preservation (p = 0.05). Use of PER(ret) resulted in significantly higher ROC as compared with PER(ant) (p < 0.001) and was comparable to results obtained with CE-preservation, which was not further improved with retrograde application. CONCLUSIONS: Celsior provides better lung preservation than does Perfadex when administered antegradely. Retrograde application of Perfadex results in significant functional improvement as compared with antegrade perfusion, which reaches the standard of Celsior protected organs. Additional in vivo experiments in combination with ultrastructural analysis are warranted to further evaluate retrograde delivery of preservation solutions, which could be used in clinical lung transplantation to further optimize current results. PMID- 11044694 TI - Transplant immunosuppression increases and prolongs transgene expression following adenoviral-mediated transfection of rat lungs. AB - BACKGROUND: Gene therapy provides the potential to modify donor organs to better withstand transplantation, but this has yet to be realized. In vivo gene transfer using adenoviral vectors has had limited success because of host immune response that induces inflammation and limits the amount and duration of transgene expression. We hypothesize that transplantation immunosuppression can attenuate the post-transfection host-immune response to allow for improved gene transfer following adenoviral-mediated transfection. METHODS: We intratracheally transfected with adenovirus containing the beta-galactosidase gene and randomized the rats to either the immunosuppression group, receiving daily cyclosporine, azathioprine, and methylprednisolone, or the control group, receiving no immunosuppression. We evaluated transgene expression and post-transfection inflammation at time points ranging from 1 day to 5 weeks. RESULTS: Following transfection, control rats showed relatively low levels of transgene expression, which rapidly decreased to non-detectable levels. In contrast, immunosuppressed rats demonstrated significantly higher levels of transgene expression overall (p < 0.00005), peaking at almost 3 times that of the control group (p < 0.02), and showing prolonged and elevated transgene expression at 5 weeks (p < 0.02). On histologic sections of the lungs, immunosuppressed rats exhibited overall lesser grades of post-transfection inflammation. CONCLUSIONS: Transplant immunosuppression provides the means to attenuate the severe immune response to adenoviral-mediated gene transfection and thereby increase and prolong transgene expression. PMID- 11044695 TI - Activities of daily living among heart transplant candidates: neuropsychological and cardiac function predictors. AB - BACKGROUND: The ability of patients to perform day-to-day tasks (e.g., medication management, dietary regulation) is an important concern of transplant teams. METHODS: We studied a clinical series of 75 heart transplant candidates and 38 controls to examine the predictive validity of demographic, neuropsychologic, and cardiac function variables to a performance-based measure of instrumental activities of daily living (IADL) capacity (i.e., Everyday Problems Test, EPT). RESULTS: Multiple regression analyses, controlling for education and race, indicated that neuropsychologic tests accounted for between 34% and 67% of the variance across IADL domains (e.g., cooking, household chores, medication management). The IADL capacity was most consistently predicted by long-standing verbal ability (Shipley Institute of Living Scale-Vocabulary, SILS-VOC) and psychomotor speed and mental flexibility (Trail Making Test-Part B, TMT-B). Similarly, SILS-VOC and TMT-B also tended to show the best operating characteristics (i.e., sensitivity, specificity, positive predictive power, negative predictive power) for detection of dependence across IADL domains. In contrast, cardiac function measures (e.g., cardiac output, mean atrial pressure) were largely unrelated to the patient's performance on the paper-and-pencil EPT task. CONCLUSIONS: Long-standing intellectual ability, and a measure of speeded information processing and mental flexibility are the best predictors of IADL capacity. PMID- 11044696 TI - Interatrial conduction of atrial tachycardia in heart transplant recipients: potential pathophysiology. AB - Surgical suture lines formed at the site of anastamosis have been considered to be electrically inert and thus present a line of block to conduction. However, a number of reports have suggested that conduction is occasionally possible across suture lines. Most of these cases have reported conduction between donor and recipient atria following cardiac transplantation. We report an illustrative case successfully treated with radiofrequency ablation, and present pathology findings that may give insight into the pathophysiology. PMID- 11044697 TI - Recurrent medulloblastoma following pediatric double-lung transplant. AB - We report a case of recurrent medulloblastoma following successful pediatric double-lung transplant for chemotherapy-induced pulmonary fibrosis. The patient had an apparent 10-year malignancy-free period prior to the transplant. This case demonstrates a potential complication of lung transplantation in individuals with prior malignancies, and questions whether patients with a history of medulloblastoma are suitable candidates for lung transplantation. PMID- 11044698 TI - Adolescents, health services, and access to care. PMID- 11044699 TI - "Reducing the Odds" at what cost: will routine testing of pregnant adolescents decrease perinatal transmission of HIV? PMID- 11044700 TI - Health care service use and sexual communication: past experience and future intention of high-risk male adolescents. AB - High-risk male adolescents were surveyed to collect data to be used to develop strategies to enhance communication with their partners about sexually transmitted diseases (STDs). Participants were concerned about acquiring an STD from their partner, and reported an increased likelihood of communicating about STD-related issues when confidential health care was available. PMID- 11044701 TI - Dating violence: a comparison of rural, suburban, and urban teens. AB - This study analyzed rural, suburban, and urban differences in teen dating violence using the 1996 Teen Assessment Project data. Teens in rural school districts were more likely to be victims of dating violence than their suburban and urban counterparts; males reported being slapped, hit, or kicked more frequently than females. The findings of this study indicate that students in rural school districts are at greater risk for participating in dating violence than suburban and urban students, with rural female students at greatest risk. PMID- 11044702 TI - Research articles published in the Journal of Adolescent Health: a two-decade comparison. AB - PURPOSE: To examine changes in subject contents and study designs of research articles published in the Journal of Adolescent Health since its inception. METHODS: A retrospective review of all research articles was conducted from selected years, ranging from 1980 through 1998. The study sample was composed of the following: original articles, case reports, brief scientific reports, international articles, fellowship forum, and health briefs. RESULTS: A total of 582 articles were evaluated. The total percentage of medical topics in research articles published in the Journal decreased from 61% in 1980-1981 to 38% in 1997 1998 (p <.01). This finding was in contrast to topics related to psychosocial issues, which increased from 23% to 50% (p <.01) over the same period. This change was largely accounted for by studies focusing on high-risk behavior. Retrospective designs, including case reports/series and chart reviews, decreased from 25% of all research articles in 1980-1981 to 9% in 1997-1998 (p <.01). The percentage of observational studies, i.e., those using cross-sectional and longitudinal designs, increased from 62% to 79% over the same period (p <.01). No changes were observed in the percentage of experimental designs, never exceeding >5% of total study designs. Finally, over the 20 years, professional background and academic departments of first authors of research broadened, with increasing contributions from nonphysicians and from non-pediatric disciplines such as psychology, public health, and nutrition. CONCLUSION: A shift in subject content of research articles from medical to psychosocial topics was observed over the past 2 decades. A shift in research designs from retrospective to cross-sectional and longitudinal was observed over the same period. The pool of authors has diversified. PMID- 11044703 TI - Breakfast consumption with and without vitamin-mineral supplement use favorably impacts daily nutrient intake of ninth-grade students. AB - PURPOSE: To assess the contribution of breakfast consumption (with and without dietary supplement intake) on total daily nutrient intake of ninth-grade students. METHODS: Twenty-four-hour recall of dietary intake was collected from a random sample of 711 ninth-grade students attending 12 Archdiocesan high schools in New Orleans, Louisiana. Analysis of variance techniques, Pearson's Chi-square statistic, and Breznahn-Shapiro method with Scheffe probabilities were used to analyze nutrient intake data, dietary adequacy, and nonorthogonal comparisons, respectively. RESULTS: Nineteen percent of 15-year-olds skipped breakfast, with more females skipping breakfast than males (23% vs. 14%, respectively). Thirty six percent of nonwhite females versus 20% white females skipped breakfast. Eleven percent of subjects took some type of dietary supplement, most commonly a multivitamin and mineral supplement. Among those who ate breakfast, average energy intake from breakfast was 437 kcal. Percentage of total daily energy intake was higher from fats and lower from carbohydrates for adolescents who skipped breakfast, compared with adolescents who consumed breakfast. The percentage of subjects consuming at least two-thirds of the Recommended Dietary Allowance was significantly lower among adolescents skipping breakfast than those consuming breakfast. CONCLUSION: Regardless of supplement use, breakfast consumption makes an important nutritional contribution to total daily intake of ninth-grade students. Encouraging breakfast consumption and healthful breakfast choices is an important step toward improving the nutritional quality of diets of this age group. PMID- 11044704 TI - Prevalence of health risk behaviors among Asian American/Pacific Islander high school students. AB - PURPOSE: [corrected] To compare the prevalence of selected risk behaviors among Asian American/Pacific Islander (AAPI) students and white, black, and Hispanic high school students in the United States. METHODS: The national Youth Risk Behavior Survey conducted in 1991, 1993, 1995, and 1997 by the Centers for Disease Control and Prevention produced nationally representative samples of students in grades 9 through 12 in all 50 states and the District of Columbia. To generate a sufficient sample of AAPI students, data from these four surveys were combined into one dataset yielding a total sample size of 55, 734 students. RESULTS: In the month preceding the survey, AAPI students were significantly less likely than black, Hispanic, or white students to have drunk alcohol or used marijuana. AAPI students also were significantly less likely than white, black, or Hispanic students to have had sexual intercourse; however, once sexually active, AAPI students were as likely as other racial or ethnic groups to have used alcohol or drugs at last intercourse or to have used a condom at last intercourse. AAPI students were significantly less likely than white, black, or Hispanic students to have carried a weapon or fought but were as likely as any of the other groups to have attempted suicide. CONCLUSIONS: A substantial percentage of AAPI students engage in risk behaviors that can affect their current and future health. Prevention programs should address the risks faced by AAPI students using culturally sensitive strategies and materials. More studies are needed to understand the comparative prevalence of various risk behaviors among AAPI subgroups. PMID- 11044705 TI - Homeless and runaway youths' access to health care. AB - PURPOSE: To describe use of health services and self-reported access to regular and emergency care by homeless adolescents and street youth. METHODS: Interviewer administered surveys addressed use of health services, availability of sources of care for emergencies, and types of care sources used. An abbreviated version of the questionnaire used for youth in shelters was used for street youth. A nationally representative sample of 640 sheltered youth and a purposive sample of 600 street youth aged 12-21 years were interviewed. All data were collected in 1992. RESULTS: Half of street youth and 36% of sheltered youth did not have a regular source of health care (p < or =.05). One-fourth of street youth and 18% of sheltered youth also reported serious health problems within the past year (p < or =.05). Street youth were more likely than sheltered youth to have used emergency treatment (36% vs. 29%; p < or =.05) and alcohol- or drug-related emergency treatment (25% vs. 13%; p < or =.05). Sheltered youth with a regular source of care were more likely to use nonemergency sites than those without a source of primary care (46% vs. 20%; p < or =.001). Few sheltered or street youth perceived shelter clinics, clinics for runaway youth, or free youth clinics to be available to meet their emergency care needs. CONCLUSIONS: Significant numbers of homeless youth did not have a regular source of health care. Those who had a regular source of care were more likely to have continuity between routine and emergency care. Integration of health services with other agencies serving youth in shelters or on the street may improve access to care for those without a routine source of care and provide better continuity for these high-risk youth. PMID- 11044706 TI - Adolescents with learning disabilities: risk and protective factors associated with emotional well-being: findings from the National Longitudinal Study of Adolescent Health. AB - PURPOSE: To identify differences in emotional well-being among adolescents with and without learning disabilities and to identify risk and protective factors associated with emotional distress. METHODS: Cross-sectional analysis of adolescent in-home interview data of the National Longitudinal Study of Adolescent Health. A total of 20,780 adolescents were included in this study of whom 1,301 were identified as having a learning disability. Initially, emotional distress, suicidal behaviors, and violence involvement were compared among those adolescents with and without learning disabilities using Student's t-test for the continuous or semicontinuous variables and Chi-square for the dichotomous variables. Subsequently, logistic regression analyses were conducted to identify which variables were most strongly associated with risk and protective factors for emotional distress. RESULTS: Adolescents with learning disabilities had twice the risk of emotional distress, and females were at twice the risk of attempting suicide and for violence involvement than their peers. While educational achievement is below that of peers, connectedness to school is comparable. So, too, is connectedness to parents. Connectedness to parents and school was identified as most strongly associated with diminished emotional distress, suicide attempts, and violence involvement among adolescents with learning disabilities. CONCLUSIONS: Given the increased association with emotional distress, suicidal attempts, and violence involvement, clinicians need to assess social and emotional as well as educational and physical functioning of these young people. We also need to be aware of the role protective factors play in the lives of young people with learning disabilities. PMID- 11044707 TI - A teenager with an annoying cough. AB - Chronic cough is a stressful condition and can lead to extensive investigations. Bronchial asthma and postnasal drip syndrome are common causes, but sometimes the origin of cough is outside the respiratory tract (1,2). Such a relatively simple test as esophageal pH probing may suggest appropriate (antireflux) therapy. PMID- 11044708 TI - Health status and risk behavior of adolescents in the north of Madrid, Spain. AB - PURPOSE: To study the health status and risk behavior in an adolescent population of Madrid, Spain. METHODS: An anonymous self-administered questionnaire was completed by 2831 pupils aged 14-20 years from urban, suburban, and rural populations in the north of Madrid. A large number of questions were from the Minnesota Adolescent Health Survey questionnaire, which included sociodemographics, health status, drug use, sexual practices, suicidal thoughts, emotional problems, and injury risk. Student's t-test, analysis of variance, and Chi-square statistics are used for analyzing associations between the different variables. RESULTS: A majority (80%) of students thought they are in good or very good health (better for males than females); 18.3% had had intercourse, the average age of the first intercourse was 15.4 years +/- 1.68 SD for males and 16.1 years +/- 1.46 SD for females. Most (73%) had used some kind of contraception. Eighty-five percent had tried alcohol, and regular use (frequently or every day) was as high as 24%. Males drank more than females (28% vs. 20%); 58.2% of the population drink only during weekends, the percentage increasing with age. Seventy percent of the sample had smoked at least once; 34. 5% reported regular tobacco use and this increased with age. Males smoked less than females (27% vs. 42%); 3.2% smoked more than 20 cigarettes/day. The consumption of illegal drugs was very low (20% had tasted cannabis and use by males was higher). Cocaine, heroin, etc. were used by less than 5%. CONCLUSION: In general, our adolescents are in good health with most problems being preventable. Sexual activity was lower than in other European and American countries. The consumption of tobacco and alcohol started early and in general was high. Our study confirms the relationship among different health-compromising behaviors. There is a need to implement health promotion and preventive programs for adolescents. PMID- 11044710 TI - Cardiology PMID- 11044709 TI - Sexual coercion among youth and young adults in Lima, Peru. AB - PURPOSE: To determine the prevalence and correlates of sexual coercion in young adults in Lima, Peru. METHODS: Lifetime sexual coercion and that at first sexual experience were studied in 629 sexually active young people, drawn from representative samples of 611 adolescents and 607 young adults. RESULTS: Almost half of the young women and a quarter of the young men in the study reported sexual coercion. In multiple logistic regression analyses, men and women who reported having been coerced at heterosexual initiation also reported more lifetime sexually transmitted diseases and a lower age at first sex than those not reporting coercion. Men who reported coercion at heterosexual initiation also reported a lower number of lifetime heterosexual partners and less sexual knowledge than men not coerced. CONCLUSION: Experiencing heterosexual initiation as coercive appears to be a marker for a riskier sexual career for both genders and for future homosexual behavior in men. PMID- 11044711 TI - Silencing genes silencing genes. PMID- 11044712 TI - Metabolic engineering of plant carotenoids. PMID- 11044713 TI - ZEITLUPE and FKF1: novel connections between flowering time and circadian clock control. PMID- 11044714 TI - Phosphorus acquisition; plant in the driver's seat! PMID- 11044715 TI - Sterols as regulators of plant embryogenesis. PMID- 11044717 TI - Fusarium control by fusarium PMID- 11044716 TI - CLV1 and CLV3: negative regulators of SAM stem cell accumulation. PMID- 11044718 TI - Proteomics of the chloroplast: experimentation and prediction. AB - New technologies, in combination with increasing amounts of plant genome sequence data, have opened up incredible experimental possibilities to identify the total set of chloroplast proteins (the chloroplast proteome) as well as their expression levels and post-translational modifications in a global manner. This is summarized under the term 'proteomics' and typically involves two-dimensional electrophoresis or chromatography, mass spectrometry and bioinformatics. Complemented with nucleotide-based global techniques, proteomics is expected to provide many new insights into chloroplast biogenesis, adaptation and function. PMID- 11044719 TI - Degradation pathway(s) of chlorophyll: what has gene cloning revealed? AB - The mechanism responsible for the degreening of plants and the degradation of chlorophyll was unclear for many years. However, recent studies have identified the colorless intermediates and helped to construct a basic pathway for degradation. After the successive removal of phytol and Mg21 from the chlorophyll molecule by chlorophyllase and 'Mg dechelatase', pheophorbide a is cleaved and reduced to yield a colorless, open tetrapyrrole intermediate. After further modifications, this is finally transported to the vacuole. Cloning the genes for chlorophyllase isozymes and the reductase should help to elucidate the physiological roles of each enzyme at a molecular level. PMID- 11044720 TI - Molecular mechanisms of self-recognition in Brassica self-incompatibility. AB - Plants have mechanisms to promote outbreeding and thereby to increase their genetic diversity. In species that are self-incompatible, self-pollen is rejected by the stigma. This mechanism has been the subject of intense study for many years and, in the past two years, significant progress has been made in identifying the genes involved in Brassica. Self-recognition involves two genes, one of which determines the male and the other the female specificity. Considerable progress has also been made on the mechanism by which self recognition leads to pollen rejection, although the delineation of all the genes involved is still not complete. PMID- 11044721 TI - Genetics and biochemistry of secondary metabolites in plants: an evolutionary perspective. AB - The evolution of new genes to make novel secondary compounds in plants is an ongoing process and might account for most of the differences in gene function among plant genomes. Although there are many substrates and products in plant secondary metabolism, there are only a few types of reactions. Repeated evolution is a special form of convergent evolution in which new enzymes with the same function evolve independently in separate plant lineages from a shared pool of related enzymes with similar but not identical functions. This appears to be common in secondary metabolism and might confound the assignment of gene function based on sequence information alone. PMID- 11044722 TI - Technical Focus:a guide to Agrobacterium binary Ti vectors. PMID- 11044723 TI - Mixed nicotinic-muscarinic properties of the alpha9 nicotinic cholinergic receptor. AB - The rat alpha9 nicotinic acetylcholine receptor (nAChR) was expressed in Xenopus laevis oocytes and tested for its sensitivity to a wide variety of cholinergic compounds. Acetylcholine (ACh), carbachol, choline and methylcarbachol elicited agonist-evoked currents, giving maximal or near maximal responses. Both the nicotinic agonist suberyldicholine as well as the muscarinic agonists McN-A-343 and methylfurtrethonium behaved as weak partial agonists of the receptor. Most classical cholinergic compounds tested, being either nicotinic (nicotine, epibatidine, cytisine, methyllycaconitine, mecamylamine, dihydro-beta erythroidine), or muscarinic (muscarine, atropine, gallamine, pilocarpine, bethanechol) agonists and antagonists, blocked the recombinant alpha9 receptor. Block by nicotine, epibatidine, cytisine, methyllycaconitine and atropine was overcome at high ACh concentrations, suggesting a competitive type of block. The present results indicate that alpha9 displays mixed nicotinic-muscarinic features that resemble the ones described for the cholinergic receptor of cochlear outer hair cells (OHCs). We suggest that alpha9 contains the structural determinants responsible for the pharmacological properties of the native receptor. PMID- 11044724 TI - Block of the alpha9 nicotinic receptor by ototoxic aminoglycosides. AB - In the present study, we report that the alpha9 nicotinic acetylcholine receptor (nAChR) expressed in Xenopus laevis oocytes is reversibly blocked by aminoglycoside antibiotics. The aminoglycosides tested blocked the alpha9 nAChR in a concentration-dependent manner with the following rank order of potency: neomycin>gentamicin>streptomycin>amikacin>kanamycin. The antagonistic effect of gentamicin was not overcome by increasing the concentration of acetylcholine (ACh), indicative of a non-competitive type of block. Blockage of ACh-evoked currents by gentamicin was found to be voltage-dependent, being more potent at hyperpolarized than at depolarized holding potentials. Furthermore, gentamicin blockage was dependent upon the extracellular Ca(2+) concentration, shown by the fact that increments in extracellular Ca(2+) significantly reduced the potency of this aminoglycoside to block the alpha9 nAChR. Possible mechanisms of blockage by the aminoglycosides are discussed. The present results suggest that the initial reversible actions of aminoglycosides at the organ of Corti, such as the elimination of the olivocochlear efferent function, are due in part to the interaction with the native alpha9-containing cholinergic receptor of the outer hair cells. PMID- 11044725 TI - Effects of diltiazem on human nicotinic acetylcholine and GABA(A) receptors. AB - Effects of the L-type calcium channel antagonist diltiazem on recombinant human GABA(A) receptor (alpha1beta2gamma2s) or on muscle (alpha1beta1deltagamma and alpha1beta1delta(epsilon)) or neuronal (alpha7 and alpha4beta2) nicotinic acetylcholine receptors expressed in Xenopus oocytes were examined using two electrode voltage-clamp. Diltiazem inhibited the function of both muscle and neuronal nicotinic receptors, but it had no effect on GABA(A) receptors. The extent of functional inhibition of nicotinic receptors depended on the receptor subtype, and the order of inhibition potency by diltiazem was alpha7>alpha4beta2 approximately alpha1beta1deltagamma approximately alpha1beta1delta(epsilon). Inhibition of alpha7 receptor function was non-competitive and voltage independent, and it occurred at concentrations far lower than those needed to inhibit (never completely) binding of (125)I-alpha-bungarotoxin to heterologously expressed alpha7 receptors in mammalian cells. Pre-incubation in diltiazem before concomitant application with acetylcholine increased inhibition of function and slowed recovery from inhibition. Verapamil, a phenylalkylamine antagonist of L type Ca(2+) channels also fully inhibited alpha7 receptor function and partially inhibited (125)I-alpha-bungarotoxin binding to alpha7 receptors, but was less potent than diltiazem. Effects on both alpha7 receptor function and (125)I-alpha bungarotoxin binding by verapamil plus diltiazem suggest separate sites for verapamil and diltiazem on alpha7 receptors. These results provide further evidence that L-type Ca(2+) channel drugs inhibit ligand-gated cationic channels and suggest that caution should be applied when using these compounds to study systems in which L-type Ca(2+) channels and ligand-gated cationic channels co exist. PMID- 11044726 TI - Characterization of the recombinant human neuronal nicotinic acetylcholine receptors alpha3beta2 and alpha4beta2 stably expressed in HEK293 cells. AB - HEK293 cells were stably transfected with the cDNAs encoding full-length human neuronal nicotinic acetylcholine receptor (nAChR) subunit combinations alpha3beta2 or alpha4beta2. [(3)H]-(+/-)Epibatidine ([(3)H]-(+/-)EPI) bound to membranes from A3B2 (alpha3beta2) and A4B2.2 (alpha4beta2) cells with K(d) values of 7.5 and 33.4 pM and B(max) values of 497 and 1564 fmol/mg protein, respectively. Concentration-dependent increases in intracellular free Ca(2+) concentration were elicited by nAChR agonists with a rank order of potency of EPI>1,1-dimethyl-4-phenylpiperazinium (DMPP)>nicotine (NIC)=suberyldicholine (SUB)>cytisine (CYT)=acetylcholine (ACh) for A3B2 cells and EPI>CYT=SUB=NIC=DMPP>ACh for A4B2.2 cells. Antagonists of nAChRs blocked NIC induced responses with a rank order of potency of d-tubocurarine (d Tubo)=mecamylamine (MEC)>dihydro-beta-erythroidine (DHbetaE) in A3B2 cells and MEC=DHbetaE>d-Tubo in A4B2.2 cells. Whole-cell patch clamp recordings indicate that the decay rate of macroscopic ACh-induced currents is faster in A3B2 than in A4B2.2 cells and that A3B2 cells are less sensitive to ACh than A4B2.2 cells. ACh currents elicited in alpha3beta2 and alpha4beta2 human nAChRs are maximally potentiated at 20 and 2 mM external Ca(2+), respectively. Our results indicate that stably expressed alpha3beta2 and alpha4beta2 human nAChRs are pharmacologically and functionally distinct. PMID- 11044727 TI - The unusual nature of epibatidine responses at the alpha4beta2 nicotinic acetylcholine receptor. AB - The identification of an equatorial frog toxin, epibatidine, as a potent non morphinic analgesic, selective for neuronal nicotinic acetylcholine receptors, provoked a marked renewal in our understanding of pain and its mechanisms. In this work we have examined the effects of epibatidine at the major brain rat alpha4beta2 nicotinic acetylcholine receptor expressed in a cell line. Fast drug applications obtained with a modified liquid filament system were used for the analyses of the currents evoked by acetylcholine, nicotine and epibatidine. Characterized by a slow onset and offset, epibatidine responses were of smaller amplitude to those evoked by acetylcholine or nicotine. About a thousand times more sensitive to epibatidine than acetylcholine, the alpha4beta2 receptor also displayed a more pronounced apparent desensitization to this compound. Finally, overnight exposure to 1 nM epibatidine failed to produce the functional upregulation observed with nicotine. These data indicate that, at the rat alpha4beta2 receptor, epibatidine acts as a partial agonist causing a pronounced inhibition of agonist evoked currents at concentrations that do not activate the receptors. PMID- 11044728 TI - Human alpha6 AChR subtypes: subunit composition, assembly, and pharmacological responses. AB - Many nicotinic acetylcholine receptor (AChR) subunits are known to be co expressed with the alpha6 subunit in neurons. Because alpha6beta4 AChRs assemble inefficiently and alpha6beta2 AChRs not at all, more complex mixtures of human subunit cDNAs were tested for their abilities to form functional AChRs when expressed in Xenopus oocytes. alpha6beta4beta3 AChRs produced the largest and most consistent responses. alpha6alpha3beta2 AChRs exhibited reduced potency for ACh and increased potency and efficacy for nicotine compared to alpha3beta2 AChRs, but similar resistance to functional inactivation after prolonged exposure to nicotine. alpha6alpha4beta2 AChRs differed little in potency or efficacy for ACh or nicotine compared to alpha4beta2 AChRs, and had similarly high sensitivity to inactivation by prolonged exposure to nicotine. Co-expression of alpha6 and beta2 cRNAs resulted in large numbers of (3)H-epibatidine binding sites in the form of large aggregates but not in functional pentameric AChRs. Co-expression of alpha6, beta2, and alpha5 resulted in assembly of some functional pentameric AChRs. Chimeras with the large extracellular domain of alpha6 and the rest from either alpha3 or alpha4 efficiently formed functional AChRs. Thus, the extracellular domain of alpha6 efficiently assembles with beta2 to form ACh binding sites, but more C-terminal domains cause difficulties in forming pentameric AChRs. Chimeric alpha6/alpha3 and alpha6/alpha4 AChRs containing either beta2 or beta4 subunits were blocked by alpha-conotoxin MII which had previously been reported to be specific for alpha3beta2 AChRs. PMID- 11044729 TI - Nicotinic acetylcholine receptor subunit mRNA expression and channel function in medial habenula neurons. AB - Relationships between nicotinic acetylcholine receptor (nAChR) channel function and nAChR subunit mRNA expression were explored in acutely isolated rat medial habenula (MHb) neurons using a combination of whole-cell recording and single cell RT-PCR techniques. Following amplification using subunit-specific primers, subunits could be categorized in one of three ways: (i) present in 95-100% cells: alpha3, alpha4, alpha5, beta2 and beta4; (ii) never present: alpha2; and (iii) sometimes present ( approximately 40% cells): alpha6, alpha7 and beta3. These data imply that alpha2 subunits do not participate in nAChRs on MHb cells, that alpha6, alpha7 and beta3 subunits are not necessary for functional channels but may contribute in some cells, and that nAChRs may require combinations of all or subsets of alpha3, alpha4, alpha5, beta2 and beta4 subunits. Little difference in the patterns of subunit expression between nicotine-sensitive and insensitive cells were revealed based on this qualitative analysis, implying that gene transcription per se may be an insufficient determinant of nAChR channel function. Normalization of nAChR subunit levels to the amount of actin mRNA, however, revealed that cells with functional channels were associated with high levels (>0.78 relative to actin; 11/12 cells) of all of the category (i) subunits: alpha3, alpha4, alpha5, beta2 and beta4. Conversely, one or more of these subunits was always low (<0.40 relative to actin) in all cells with no detectable response to nicotine. Thus the formation of functional nAChR channels on MHb cells may require critical levels of several subunit mRNAs. PMID- 11044730 TI - Cloning and heterologous expression of Dalpha4, a Drosophila neuronal nicotinic acetylcholine receptor subunit: identification of an alternative exon influencing the efficiency of subunit assembly. AB - A neuronal nicotinic acetylcholine receptor (nAChR) subunit, Dalpha4, has been identified and cloned from the fruit fly Drosophila melanogaster, together with several alternatively spliced transcripts. Intron-exon boundaries within the gene encoding Dalpha4 (nAcRalpha-80B) have been identified by comparison of cDNA and genomic sequence data. The influence of amino acids encoded by alternatively spliced exons upon nicotinic radioligand binding and subunit-subunit co-assembly has been examined by heterologous expression in Drosophila S2 cells. The efficiency of subunit assembly has been shown to be influenced by amino acids surrounding the highly conserved 15 amino acid cysteine-loop motif within the N terminal extracellular domain of the nAChR Dalpha4 subunit. Extensive use has been made of publicly available data determined by the Berkeley Drosophila Genome Project (BDGP). This includes expressed sequence tag (EST) data as well as whole embryo in situ hybridisation and polytene chromosome in situ hybridisation data. BDGP in situ hybridisation data suggests that the Dalpha4 mRNA is expressed within Drosophila brain and ventral nerve cord and demonstrates that the gene encoding this nAChR subunit is located at position 80B on chromosome 3. The relationship between Dalpha4 and other previously cloned nAChR subunits has been examined and the implications for the nomenclature of insect nAChRs is discussed. PMID- 11044731 TI - Synergistic transcriptional activation by Sox10 and Sp1 family members. AB - Neuronal nicotinic acetylcholine receptors (nAChR) are expressed at specific times during development and in discrete neuronal populations. Transcriptional regulation of the receptor genes clearly plays a key role in the molecular pathway underlying the expression of these critical synaptic components. In an effort to understand this regulation, we focus upon the genes encoding three receptor subunits: alpha3, alpha5 and beta4. These subunits are genomically clustered and constitute the predominant nAChR subtype expressed in the peripheral nervous system. We and others demonstrated that the general transcription factors, Sp1 and Sp3, can transactivate the promoter of each subunit gene. Further, we showed that the regulatory factor Sox10 transactivates the alpha3 and beta4 promoters and does so in a cell-type-specific manner. Interestingly, the Sp- and Sox10-binding sites on the beta4 promoter are located immediately adjacent to each other, raising the possibility that the two sets of factors functionally interact to regulate receptor gene expression. Consistent with this hypothesis, we demonstrated that the proteins can directly interact. Here, we extend these observations and show that Sox10 and the Sp factors functionally interact, leading to synergistic transcriptional activation in a cholinergic cell line. Finally, evidence for the existence of cell-type-specific co-regulators for Sp1 and Sox10 is presented. PMID- 11044732 TI - Expression and function of striatal nAChRs differ in the flinders sensitive (FSL) and resistant (FRL) rat lines. AB - Rats of Flinders Sensitive (FSL) and Flinders Resistant lines (FRL) differ in their susceptibility to physiological and associated behavioral responses elicited by nicotine. In the present study, we measured dopamine (DA) content in striatal dialysates to investigate the sensitivity of FSL and FRL rats to nicotine delivered locally through a microdialysis probe placed in the striatum. We also measured the expression density of striatal high-affinity nicotinic acetylcholine receptors (nAChRs), and that of mRNAs encoding for alpha3, alpha4, alpha7 and beta2 nAChR subunits in both lines. The DA content of dialysates was measured before and after a 1-min perfusion of nicotine (6, 10 or 20 nmoles/min) and the resulting DA increase was taken as a measure of the alkaloid's intrinsic activity for nAChRs involved in the release of DA. The nicotine-induced increase of striatal DA release was greater in FSL than in FRL rats for all concentrations of nicotine, suggesting that the intrinsic activity of nicotine was greater in the FSL than in the FRL rats. This was further supported by our finding that the density of high-affinity nAChRs in the striatum of FSL rats was 44% greater than in the FRL rats, whereas affinity (K(D)) was virtually the same in the two lines of rats. Also the expression of mRNAs encoding for alpha(4), alpha(7), and beta(2) subunits in the striatum was greater in FSL than in FRL rats (attomol/microg total RNA, alpha(4):98+/-10 vs. 77+/-7; alpha(7):279+/-16 vs. 184+/-16; beta(2):310+/-19 vs. 201+/-12). We hypothesize that the difference in nicotine-induced DA release in the striatum of FSL and FRL rats depends on the difference in nAChR subunit expression in the striatum between the two lines. The Flinders rats could be used as a model for nicotine self-administration studies to evaluate the susceptibilities of FSL and FRL rats to nicotine dependence. PMID- 11044733 TI - Nicotinic-agonist stimulated (86)Rb(+) efflux and [(3)H]epibatidine binding of mice differing in beta2 genotype. AB - Nicotinic acetylcholine receptor function and binding was measured in 12 brain regions from mice differing in beta2 subunit expression. Function was measured by on-line detection of (86)Rb(+) efflux stimulated under conditions that measure two pharmacologically distinct nicotinic responses: (1) stimulation with 10 microM nicotine, a response that is relatively sensitive to inhibition by the antagonist, dihydro-beta-erythroidine (DHbetaE); and (2) stimulation with 10 microM epibatidine in the presence of 2 microM DHbetaE, a response that is relatively resistant to inhibition by DHbetaE. Deletion of the beta2 subunit profoundly reduced both DHbetaE-sensitive and -resistant (86)Rb(+) efflux in each brain region and essentially eliminated activity in regions such as cerebral cortex and thalamus. However, residual activity was observed in regions such as olfactory bulbs and inferior colliculus. [(3)H]Epibatidine binding was measured under conditions that allow estimation of both high- and low-affinity sites. High affinity sites sensitive to inhibition by the nicotinic agonist, cytisine, were virtually eliminated in every region by the beta2 null mutation. In contrast, only a subset of the high-affinity sites insensitive to inhibition by cytisine were eliminated in beta2 null mutants, suggesting receptor heterogeniety. Similarly, low affinity [(3)H]epibatidine binding was heterogeneous in that a fraction of the sites required the beta2 subunit. Many remaining sites were sensitive to inhibition by alpha-bungarotoxin indicating that a subset of the low affinity [(3)H]epibatidine binding are of the alpha7* subtype. Distinct regional variation was observed among the 12 brain regions. These studies confirm important roles for beta2-containing receptors in mediating pharmacologically distinct functions and as components of several identifiable binding sites. PMID- 11044734 TI - The nicotinic modulation of [(3)H]D-aspartate outflow in primary cultures of rat neocortical neurons: effect of acute and long term nicotine treatment. AB - The effect of nicotine 1 nM-10 microM on the efflux of [(3)H]D-aspartate was tested in primary cultures of rat cortical neurons kept at rest and subjected to electrical field stimulation. Two trains of pulses at 20 Hz for 20 s were applied at the 60th (St(1)) and 90th (St(2)) min of perfusion. The drug slightly and transiently increased the efflux of resting cells while, when given during St(2), it greatly enhanced the electrically evoked efflux estimated as St(2)/St(1) ratio, EC(50) being 107 nM. The nicotinic receptors (nAChR) giving rise to this positive modulation were partly mecamylamine- and partly alpha-bungarotoxin sensitive. They appeared to be located at the nerve endings since nicotine facilitation was only slightly prevented by tetrodotoxin during depolarisation with 15 mM KCl. Pretreatment with glutamate antagonists did not reveal any interaction between nAChR and ionotropic glutamate receptors. Membrane glutamate carrier involvement in the nicotine effect was ruled out. Long-term treatment with nicotine 1 microM (from the 3rd-4th to the 8th-9th day in vitro) reduced the maximal response to the drug but shifted its threshold concentration to the left (from 10 nM to 1 nM), leaving the contribution of the two receptor subtypes unchanged. Reduced responsiveness to nicotine was also evident in long-term treated cerebellar granule cells. In conclusion, presynaptic nAChR's, both containing and lacking alpha(7) subunits, can contribute to enhance the glutamatergic secretion in primary cultures of rat cortical neurons, chiefly during electrical stimulation. PMID- 11044735 TI - Lobeline inhibits nicotine-evoked [(3)H]dopamine overflow from rat striatal slices and nicotine-evoked (86)Rb(+) efflux from thalamic synaptosomes. AB - The present study evaluated the interaction of lobeline with neuronal nicotinic acetylcholine receptors using two in vitro assays, [(3)H] overflow from [(3)H]dopamine ([(3)H]DA)-preloaded rat striatal slices and (86)Rb(+) efflux from rat thalamic synaptosomes. To assess agonist interactions, the effect of lobeline was determined and compared to S(-)-nicotine. To assess antagonist interactions, the ability of lobeline to inhibit the effect of S(-)-nicotine was determined. Both S(-)-nicotine (0.1-1 microM) and lobeline (>1.0 microM) evoked [(3)H] overflow from superfused [(3)H]DA-preloaded striatal slices. However, lobeline evoked [(3)H] overflow is mecamylamine-insensitive, indicating that this response is not mediated by nicotinic receptors. Moreover, at concentrations (<1.0 microM) which did not evoke [(3)H] overflow, lobeline inhibited S(-)-nicotine (0.1-10 microM)-evoked [(3)H] overflow, shifting the S(-)-nicotine concentration-response curve to the right. S(-)-Nicotine (30 nM-300 microM) increased (EC(50) value=0.2 microM) (86)Rb(+) efflux from thalamic synaptosomes. In contrast, lobeline (1 nM 10 microM) did not evoke (86)Rb(+) efflux, and the lack of intrinsic activity indicates that lobeline is not an agonist at this nicotinic receptor subtype. Lobeline completely inhibited (IC(50) value=0.7 microM) (86)Rb(+) efflux evoked by 1 microM S(-)-nicotine, a concentration which maximally stimulated (86)Rb(+) efflux. Thus, the results of these in vitro experiments demonstrate that lobeline inhibits the effects of S(-)-nicotine, and suggest that lobeline acts as a nicotinic receptor antagonist. PMID- 11044736 TI - Assessment of nicotinic acetylcholine receptor-mediated release of [(3)H] norepinephrine from rat brain slices using a new 96-well format assay. AB - The study of the modulatory effects of nicotinic acetylcholine receptor (nAChR) agonists on neurotransmitter release from tissue slices has been hampered by laborious and limiting superfusion techniques. A new methodology was developed utilizing 96-well filter plates. This new method produced comparable results to previously published data, yet expanded throughput to permit more complete pharmacological characterization. Rat brain slices, preloaded with [(3)H] norepinephrine ([(3)H]-NE), were distributed onto 96-well filter plates. Following a 5 min preincubation, the slices were incubated for 5 min with nicotinic agonists or antagonists. (-)-Nicotine (NIC) and 1,1-dimethyl-4 phenylpiperazine (DMPP) evoked release of [(3)H]-NE from a number of brain regions and spinal cord, with the highest response seen in the hippocampus. Concentration-response curves revealed a rank order of potency of (+/-) epibatidine>>anatoxin-a>A-85380>DMPP=NIC=(-)-cytisine in the hippocampus, thalamus, and frontal cortex. EC(50) values were approximately 0.005, 0.2, 1, 5, 5 and 5 microM, respectively. Concentration-inhibition curves of nicotine evoked [(3)H]-NE release from hippocampal and thalamic slices resulted in a rank order of potency of mecamylamine>hexamethonium>d-tubocurare>DHbetaE. Schild analysis revealed apparent noncompetitive antagonism of [(3)H]-NE release from hippocampus by mecamylamine, hexamethonium, and DHbetaE. In contrast, DHbetaE antagonism of [(3)H]-dopamine release from striatal slices using a similar methodology was competitive. PMID- 11044737 TI - Characterization of nicotinic acetylcholine receptor-mediated [(3)H]-dopamine release from rat cortex and striatum. AB - The objective of this study was to use a new high throughput method to compare nicotinic acetylcholine receptor (nAChR)-mediated [(3)H]-dopamine (DA) release from slices of rat striatum and cortex. (-)Nicotine, (-)-cytisine, 1,1-dimethyl-4 phenyl-piperazinium (DMPP), and (+/-)-epibatidine evoked release of striatal [(3)H]-DA with pEC(50) values of 6.7, 8.25, 5.11, and 9.08, respectively. The same agonists evoked release of cortical [(3)H]-DA with pEC(50) values of 6.98, 8.06, 5.58, and 9.59. Relative to (-)-nicotine, (-)-cytisine was a partial agonist in both tissues. In contrast, the maximal response evoked by DMPP differed between the two tissues. The rank order of potency for antagonists to block DA release was the same (mecamylamine (Mec)>dihydro-beta-erythroidine (DHbetaE)>hexamethonium (Hex)>D-tubocurarine (D-TC)); however, the pIC(50) values varied between the two regions. Whereas Mec potently antagonized (-)-nicotine evoked DA release similarly from striatum and cortex, with pIC(50) values of 6.07 and 6.53 respectively, the values obtained for DHbetaE, D-TC and Hex differed. Additionally, the present study was able to distinguish exocytotic vesicular mediated from transporter-mediated DA release, by altering temperature of the incubation and exclusion of calcium. Assays carried out under these conditions indicate that approximately 60% of nicotine-evoked cortical DA release was likely mediated through the DA transporter. In contrast, under the same conditions only 15%-20% of striatal release appeared to be transporter-mediated. We conclude that the relative contributions of the mechanisms by which (-)-nicotine evokes DA release differ between striatum and cortex. In addition, the data suggest that the subtypes of nAChRs involved in regulating [(3)H]-DA release may be somewhat different in the two tissues. PMID- 11044738 TI - Nicotinic receptors co-localize with 5-HT(3) serotonin receptors on striatal nerve terminals. AB - Nicotinic acetylcholine receptors and 5-HT(3) serotonin receptors are present on presynaptic nerve terminals in the striatum, where they have been shown to be involved in the regulation of dopamine release. Here, we explored the possibility that both receptor systems function on the same individual nerve terminals in the striatum, as assessed by confocal imaging of synaptosomes. On performing sequential stimulation, nicotine (500 nM) induced changes in [Ca(2+)](i) in most of the synaptosomes ( approximately 80%) that had previously responded to stimulation with the 5-HT(3) receptor agonist m-chlorophenylbiguanide (mCPBG; 100 nM), whereas mCPBG induced [Ca(2+)](i) responses in approximately half of the synaptosomes that showed responses on nicotinic stimulation. The 5-HT(3) receptor specific antagonist tropisetron blocked only the mCPBG-induced responses, but not the nicotinic responses on the same synaptosomes. Immunocytochemical staining revealed extensive co-localization of the 5-HT(3) receptor with the alpha4 nicotinic receptor subunit on the same synaptosomes, but not with the alpha3 and/or alpha5 subunits. Immunoprecipitation studies indicate that the 5-HT(3) receptor and the alpha4 nicotinic receptor subunit do not interact on the nerve terminals. The presence of nicotinic and 5-HT(3) receptors on the same presynaptic striatal nerve terminal indicates a convergence of cholinergic and serotonergic systems in the striatum. PMID- 11044739 TI - Synaptic interactions regulate gephyrin expression in avian cholinergic neurons in vivo. AB - Our recent studies of chick parasympathetic ciliary ganglion (CG) neurons demonstrate a unique postsynaptic receptor microheterogeneity - under one presynaptic terminal, excitatory nicotinic acetylcholine receptor (nAChR) clusters and separate inhibitory glycine receptor (GlyR) clusters coexist in distinct membrane microregions. Gephyrin, a peripheral membrane protein that is required for GlyR clustering at synapses in the rodent central nervous system, is also expressed in chick CG neurons where it codistributes with GlyRs, but not nAChRs. We now extend these findings by characterizing the regulation of gephyrin expression in chick CG neurons in vivo. We show that developmental increases in gephyrin transcript levels occur during pre- and postganglionic synapse formation. The increases are induced by both innervation and target tissue interactions, with the target tissues having the greater regulatory influence. The time course of the developmental rise in gephyrin mRNA levels most closely resembles that reported for functional GlyR expression, but not that of functional nAChRs nor GABA(A) receptors. We also demonstrate that gephyrin is concentrated in the postsynaptic density of a subset of synapses on both the ciliary and choroid neurons in the CG and is stably expressed from embryonic to adult stages. Altogether, our results suggest that gephyrin is a synapse organizing molecule that functions to localize GlyRs, but not nAChRs, to discrete postsynaptic membrane microregions in chick CG neurons in vivo. PMID- 11044740 TI - Cluster formation of alpha7-containing nicotinic receptors at interneuronal interfaces in cell culture. AB - Nicotinic receptors containing the alpha7 gene product are among the most abundant in the nervous system. Because of their widespread distribution and high relative permeability to calcium, the receptors regulate a diverse array of cellular events. On chick ciliary neurons the receptors are concentrated on somatic spines folded into discrete mats on the cell body and are overlaid by a large presynaptic calyx. The receptors co-localize with filamentous actin and the actin-associated protein drebrin which are concentrated in the spines. We show here that embryonic ciliary ganglion neurons grown in dissociated cell culture express and concentrate the receptors in large clusters or plaques that form at interneuronal interfaces between small clumps of neurons. The receptors resist detergent extraction even after disruption of the actin cytoskeleton, suggesting the importance of additional molecular mechanisms determining receptor location. The cell adhesion molecules N-CAM and N-cadherin are concentrated at the receptor plaques and may influence plaque stability. Although ciliary neurons do not normally contact each other in vivo, they do so in culture and may mimic interactions normally occurring between calyx and soma in vivo. As a result the cultures may prove useful for identifying components shaping development of postsynaptic specializations on neurons. PMID- 11044741 TI - Optical measurements of presynaptic nicotinic effects. AB - We have used the styryl dye FM 2-10 to monitor changes in synaptic activity mediated by nicotinic acetylcholine receptors (AChRs) on cultured hippocampal neurons. We show that both 100 microM ACh and nicotine at 20 microM causes a significant increase in staining intensities of presynaptic boutons in the presence of 0.5 microM tetrodotoxin (TTX). This effect of nicotine is blocked by d-tubocurarine. Interestingly, nicotine also had a long-lasting effect on high potassium-induced staining of boutons. These results suggest that nicotine can have significant and sustained effects on synaptic efficacy in cultured hippocampal neurons. PMID- 11044742 TI - Facilitation of glutamatergic neurotransmission by presynaptic nicotinic acetylcholine receptors. AB - The profiles of presynaptic facilitation of glutamate release as elicited by nicotine and acetylcholine were compared in two limbic pathways recapitulated in vitro. At synapses of medial habenula (MHN) and interpeduncular nucleus (IPN) neurons, application of nicotine increased the frequency of TTX-resistant, spontaneous postsynaptic currents (SSCs) by an average of 5-fold. In contrast, the average increase in SSC frequency elicited by nicotine was more than 120 fold at synapses of olfactory bulb (OB) and amygdala neurons. At both preparations, pulses of ACh caused presynaptic facilitation that lasted longer than that elicited by nicotine. The subunit composition of presynaptic nAChRs may contribute to the different profiles of facilitation observed. The large magnitude, fast kinetics, and alpha-bungarotoxin sensitivity of facilitation observed at OB-amygdala synapses is consistent with participation of alpha7-type nAChRs. As subunit-selective deletion of alpha5 or alpha7 altered the profile of nicotine-elicited facilitation at MHN-IPN synapses, presynaptic nAChRs at MHN-IPN synapses appear to be more complex. Such heteromeric combinations of nAChRs may contribute to the lower magnitude and slower kinetics of presynaptic facilitation at MHN-IPN synapses. Calcium influx through either voltage-gated calcium channels or directly through presynaptic alpha7-containing nAChRs is sufficient to support nicotine-elicited facilitation of glutamate release. Resultant increases in intracellular calcium may further modulate presynaptic nAChR activity in a subunit-composition dependent manner. PMID- 11044743 TI - Nicotine at concentrations found in cigarette smokers activates and desensitizes nicotinic acetylcholine receptors in CA1 interneurons of rat hippocampus. AB - Behavioral effects of cigarette smoking are attributed to the interactions of nicotine with brain nicotinic acetylcholine receptors (nAChRs). However, the mechanisms by which nAChR function in developing and mature brain is affected by a smoker's level of nicotine (50-500 nM) remain unclear. Thus, the objective of this study was to determine the concentration- and time-dependent effects of nicotine on alpha7 and alpha4beta2 nAChRs, the two major brain subtypes, natively expressed in CA1 interneurons of rat hippocampal slices. Only at concentrations > or =5 microM did nicotine (applied for 6-60 s) elicit action potentials or measurable whole-cell currents (EC(50)=158 microM) in stratum radiatum interneurons that express alpha7 nAChRs. Continuous exposure for 10-15 min of the neurons to nicotine (0.5-2.5 microM) inhibited alpha7 nAChR-mediated currents (IC(50)=640 nM) evoked by choline (10 mM). Nicotine (> or =0.125 microM) applied to the neurons for 1-5 min induced slowly desensitizing whole-cell currents (EC(50)=3.2 microM) in stratum lacunosum moleculare interneurons; this effect was mediated by alpha4beta2 nAChRs. Also via activation of alpha4beta2 nAChRs, nicotine (0.125-0.5 microM) increased the frequency and amplitude of GABAergic postsynaptic currents (PSCs) in stratum radiatum interneurons. However, exposure of the neurons for 10-15 min to nicotine (0.25-0.5 microM) resulted in desensitization of alpha4beta2 nAChRs. It is suggested that nanomolar concentrations of nicotine after acute intake suppress inhibitory inputs to pyramidal cells through a disinhibitory mechanism involving activation of alpha4beta2 nAChRs and desensitization of alpha7 nAChRs, and after chronic intake leads to up-regulation of both receptor subtypes via desensitization. These findings have direct implications to the actions of nicotine in cigarette smokers. PMID- 11044744 TI - The opioid antagonist naltrexone inhibits activity and alters expression of alpha7 and alpha4beta2 nicotinic receptors in hippocampal neurons: implications for smoking cessation programs. AB - This study was designed to investigate whether naltrexone, an opioid antagonist that has been evaluated clinically as a co-adjuvant in smoking cessation programs, affects function and expression of neuronal nicotinic receptors (nAChRs). Whole-cell current recordings from rat hippocampal neurons in culture and in slices demonstrated that alpha7 nAChRs can be inhibited non-competitively by naltrexone (IC(50) approximately 25 microM). The voltage dependence of the effect suggested that naltrexone acts as an open-channel blocker of alpha7 nAChRs. Naltrexone also inhibited activation of alpha4beta2 nAChRs in hippocampal neurons; however its IC(50) was higher ( approximately 141 microM). At a concentration as high as 300 microM (which is sufficient to block by 100% and 70% the activity of alpha7 and alpha4beta2 nAChRs, respectively), naltrexone had no effect on kainate and AMPA receptors, blocked by no more than 20% the activity of NMDA and glycine receptors, and reduced by 35% the activity of GABA(A) receptors. A 3-day exposure of cultured hippocampal neurons to naltrexone (30 microM) or nicotine (10 microM, a concentration that fully desensitized alpha7 nAChRs) resulted in a 2-fold increase in the average amplitude of alpha7 nAChR-subserved currents. Naltrexone did not augment the maximal up-regulation of alpha7 nAChRs induced by nicotine, indicating that both drugs act via a common mechanism. In addition to increasing alpha7 nAChRs-mediated responses per neuron, nicotine increased the number of neurons expressing functional non-alpha7 nAChRs (probably alpha4beta2 nAChRs); this effect was blocked by naltrexone (0.3 and 30 microM). Therefore, naltrexone may affect dependence on cigarette smoking by differentially altering function and expression of alpha7 and alpha4beta2 nAChRs in the central nervous system. PMID- 11044745 TI - Nicotinic receptors on hippocampal cultures can increase synaptic glutamate currents while decreasing the NMDA-receptor component. AB - Activation of presynaptic nicotinic acetylcholine receptors (nAChRs) can enhance the release of glutamate from synapses in hippocampal slices and cultures. In hippocampal cultures making autaptic connections, rapid application of a high concentration of nicotine activated presynaptic, postsynaptic, and somatic nAChRs, which consequently enhanced the amplitude of evoked excitatory postsynaptic currents (eEPSCs) mediated by glutamate receptors. The increased eEPSC amplitudes arose from enhanced glutamate release caused by presynaptic nAChRs (Radcliffe and Dani, 1998, Journal of Neuroscience 18, 7075). The same whole-cell nicotine applications that enhanced non-NMDA eEPSCs often decreased the NMDA-receptor component of the eEPSCs. Furthermore, whole-cell activation of nAChRs by nicotine selectively reduced the amplitude of the whole-cell NMDA receptor currents without affecting the non-NMDA receptor currents. The inhibition by nicotine was prevented by the alpha7-specific antagonist, methyllycaconitine, and required the presence of extracellular Ca(2+). The calmodulin antagonist fluphenazine prevented inhibition of the NMDA-receptor current by nAChR activity, suggesting that a Ca(2+)-calmodulin-dependent process mediated the effect of nicotine. Our results indicate that activation of nAChRs can modulate glutamatergic synapses in several ways. Presynaptic nAChR activity enhances synaptic transmission by increasing transmitter release. Additionally, somatic or postsynaptic nAChRs can initiate a Ca(2+) signal that can act via calmodulin to reduce the responsiveness of NMDA receptors. PMID- 11044746 TI - Ventral hippocampal alpha4beta2 nicotinic receptors and chronic nicotine effects on memory. AB - Chronic nicotine administration has been repeatedly shown to facilitate working memory function in rats on the radial-arm maze. The critical neural mechanisms for this effect are still being discovered. The nicotinic nature of the chronic nicotine induced memory improvement is supported by the finding that it is blocked by chronic mecamylamine co-infusion. The hippocampus also appears to be critically important. Hippocampal ibotenic acid lesions block the effect. Within the hippocampus, we have found that the alpha4beta2 nicotinic receptor subtype is involved in memory functioning. Acute ventral hippocampal infusions of the alpha4beta2 nicotinic antagonist dihydro-beta-erythroidine (DHbetaE) significantly decreased working memory performance in the radial-arm maze. The aim of the current study was to determine the importance of alpha4beta2 receptors within the ventral hippocampus for the memory enhancing effects of chronic nicotine treatment. Adult female Sprague-Dawley rats were trained on the 8-arm radial maze and were cannulated bilaterally in the ventral hippocampus. Osmotic minipumps administering chronic nicotine at a rate of 5 mg per kg per day were also implanted in the nicotine treatment rats. Control rats received saline-only minipumps. For a period of 4 weeks after surgery, each rat received bilateral hippocampal infusions of 0, 2, 6 and 18 microg per side of DHbetaE and tested for memory performance on the radial-arm maze. Radial-arm maze choice accuracy was impaired by acute hippocampal DHbetaE infusion in a dose-related fashion. This acute hippocampal DHbetaE-induced choice accuracy impairment was eliminated by chronic systemic nicotine infusion. Chronic nicotine in combination with acute vehicle hippocampal infusion was not seen to alter choice accuracy. Response latency was not found to be altered by acute hippocampal DHbetaE in the absence of chronic nicotine administration, but it did attenuate the response latency reduction induced by chronic nicotine infusion. Wet dog shakes were not found to be affected by hippocampal DHbetaE when given without chronic nicotine. Wet dog shakes were significantly increased by chronic nicotine infusion. Intra hippocampal DHbetaE significantly potentiated this effect. The results from the current study reinforce the hypothesis that ventral hippocampal alpha4beta2 nicotinic receptors are important for memory function. These receptors may also have a role to play in the development of other aspects of behavior associated with chronic nicotine treatment. PMID- 11044747 TI - Fear conditioning and latent inhibition in mice lacking the high affinity subclass of nicotinic acetylcholine receptors in the brain. AB - Nicotine can enhance performance in several tests of cognition but the specific nicotinic receptor subtypes mediating these effects are largely unknown. Knock out mice lacking the beta2 subunit of the nicotinic receptor were evaluated in fear conditioning and latent inhibition tasks to begin to determine which receptor subtypes mediate the cognitive effects of nicotine. Young (2-4 months) knock out and wild type mice did not differ in either contextual or tone conditioned fear, but aged (9-20 months) knock out males were impaired in freezing to both context and tone compared to aged wild type males. No differences in fear conditioning were observed between aged knock out and wild type females. Latent inhibition of fear to a pre-exposed tone, as measured by behavioral freezing, was also assessed. Both knock out and wild type mice displayed similar levels of latent inhibition, although overall levels of freezing were lower in knock out mice. These results support a previous study showing spatial learning deficits in aged beta2 subunit knock out mice [EMBO J. 18 (1999) 1235] and suggest that performance of other cognitive tasks may not be influenced by absence of beta2 subunit-containing receptors. PMID- 11044748 TI - The antinociceptive effects of alpha7 nicotinic agonists in an acute pain model. AB - Nicotinic receptors have been found to play a role in modulating pain transmission in the CNS. Activation of cholinergic pathways by nicotine and nicotinic agonists has been shown to elicit antinociceptive effects in a variety of species and pain tests. The involvement of alpha(7) nicotinic receptors in nicotinic analgesia was assessed after spinal (i.t.) and intraventricular (i.c.v.) administration in mice. Dose-dependent antinociceptive effects were seen with the alpha(7) agonist choline after spinal and supraspinal injection using the tail-flick test. Furthermore, alpha(7) antagonists MLA and alpha-BGTX significantly blocked the effects of choline. Dihydro-beta-erythroidine and mecamylamine failed to block choline-induced antinociception. These results strongly support the involvement of alpha(7) subunits in choline's antinociceptive effects. DMXB and 4-OH-DMXB, partial alpha(7) agonists, failed to elicit a significant antinociceptive effect. However, they blocked choline induced antinociception in a dose-dependent manner following i.t. injection. This antagonism is probably related to their partial agonistic properties of the alpha(7) receptors. These studies suggest that activation of alpha(7) receptors in the CNS elicits antinociceptive effects in an acute thermal pain model. PMID- 11044749 TI - Selective neurotoxic effects of nicotine on axons in fasciculus retroflexus further support evidence that this a weak link in brain across multiple drugs of abuse. AB - When administered continuously for several days at relatively low plasma levels, a variety of drugs of abuse with strong dopaminergic actions induce degeneration in axons traveling from the lateral habenula through the sheath of fasciculus retroflexus to midbrain monoaminergic nuclei. With some of these drugs, such as cocaine, this is virtually the only degeneration induced in brain. Nicotine given continuously also selectively induces degeneration in fasciculus retroflexus, but in the other half of the tract: the cholinergic axons running from medial habenula in the core of the tract to the interpeduncular nucleus. Fasciculus retroflexus appears to be a weak link in brain for diverse drugs of abuse when administered incessantly for several days. Alterations in this tract would be predicted to be especially important for the genesis of the symptomatology which develops during drug binges, residual effects of such binges, and the processes underlying relapse. PMID- 11044750 TI - The alpha7 nicotinic acetylcholine receptor subtype mediates nicotine protection against NMDA excitotoxicity in primary hippocampal cultures through a Ca(2+) dependent mechanism. AB - Neuronal nicotinic acetylcholine receptors (nAChR) have been suggested to play a role in a variety of modulatory and regulatory processes, including neuroprotection. Here we have characterized the neuroprotective effects of nicotine against an excitotoxic insult in primary hippocampal cultures. Exposure of hippocampal neurons to 200 microM NMDA for 1 h decreased cell viability by 25+/-5%, an effect blocked by NMDA receptor antagonists. Nicotine (10 microM) counteracted the NMDA-induced cell death when co-incubated with NMDA or when present subsequent to the NMDA treatment. Nicotine protection was prevented by 1 microM MLA, confirming that it was mediated by nAChR, and by 1 microM alpha bungarotoxin, demonstrating that the alpha7 nAChR subtype was responsible. Both the NMDA evoked neurotoxicity and nicotine neuroprotection were Ca(2+)-dependent. In Fura-2-loaded hippocampal neurons, nicotine (10 microM) and NMDA (200 microM) acutely increased intracellular resting Ca(2+) from 70 nM to 200 and 500 nM, respectively. Responses to NMDA were unaffected by the presence of nicotine. (45)Ca(2+) uptake after a 1 h exposure to nicotine or NMDA also demonstrated quantitative differences between the two drugs. This study demonstrates that the alpha7 subtype of nAChR can support neuronal survival after an excitotoxic stimulus, through a Ca(2+) dependent mechanism that operates downstream of NMDA receptor activation. PMID- 11044751 TI - Nicotine-induced fos expression in the pedunculopontine mesencephalic tegmentum in the rat. AB - The aim of this study was to assess the effects of a single dose of nicotine (NIC, 0.3 or 1.0 mg/kg, s.c.), after survival times of 30, 60 or 120 min, on immediate early gene expression in the pedunculopontine mesencephalic tegmentum (PMT), using Fos-immunocytochemistry. Either doses of NIC strongly increased Fos immunoreactivity in both the pedunculopontine tegmental nucleus (PPTg) and the laterodorsal tegmental nucleus (LDTg), as compared to the saline controls, at 30 min and 60 min. In comparison, the effects of NIC-induced Fos expression in the caudate-putamen (CP) were not as strong as the ones observed in the PPTg and LDTg. In fact, at 30 min the 0.3 mg/kg dose of NIC did not induce Fos-expression, unlike the PPTg and LDTg. The CP response was more noticeable in the mediodorsal than in the laterodorsal region. Double-labelling studies using Fos immunoreactivity and NADPH-diaphorase histochemistry for cholinergic cells in the PPTg and LDTg revealed that, in general, cholinergic neurons had Fos negative nuclei, although double-labelled neurons were occasionally seen in the PPTg. In conclusion, systemically administered NIC activates the neuronal population of the PPTg and the LDTg possibly by directly targeting nicotinic receptors that may be located in non-cholinergic neurons. We postulate that activation of these non cholinergic neurons modulates the activity of cholinergic cells in the PMT, which in turn may alter dopamine release in the mesolimbic system. PMID- 11044752 TI - [(3)H]Nicotine binding in peripheral blood cells of smokers is correlated with the number of cigarettes smoked per day. AB - The principal sites for biological action of tobacco products are thought to be the nicotinic acetylcholine receptors (nAChR). Nicotinic receptor subunit genes, therefore, represent an important gene family for study in nicotine addiction. They are localized in both brain and in the periphery. In brain these receptors appear to function as modulators of synaptic transmission; the function of peripheral receptors is not known. Nicotinic receptor levels in human brain are regulated by smoking in a dose-dependent manner. In peripheral blood, nicotinic receptors are present on both lymphocytes and polymorphonuclear cells (PMN). We have compared [(3)H]nicotine binding in PMN isolated from smokers and non smokers. [(3)H]nicotine binding was increased in smokers and was correlated, as in brain, with tobacco use. Expression of both mRNA and protein in lymphocytes and PMN, for a subset of nicotinic receptor subunits, suggests that these cell types contain both alpha4beta2 and alpha3beta4 receptors. PMID- 11044753 TI - Alpha and beta nicotinic acetylcholine receptors subunits and synaptophysin in putamen from Parkinson's disease. AB - It is well established that nicotinic receptors in the mammalian striatum are involved in modulation of the release of several neurotransmitters, including dopamine. In addition, nicotinic receptors with high affinity for agonists have generally been found to be reduced in the striatum in Parkinson's disease. In the present study antibodies have been used to examine which subunits contribute to the striatal nicotinic receptor loss in Parkinson's disease, and whether the reduction in [(3)H]nicotine binding correlates with synaptic loss. Autopsy tissue from the putamen of 12 Parkinson's disease cases and 12 age-matched control subjects was analysed by immunoblotting using antibodies against recombinant peptides specific for alpha3, alpha4, alpha7, beta2 and beta4 nicotinic acetylcholine receptor (nAChR) subunits and the synaptic marker synaptophysin, in conjunction with assessment of [(3)H]nicotine binding by autoradiography. The data indicate that there is no loss of alpha3, alpha4, alpha7 and beta2 immunoreactivity in the putamen in Parkinson's disease, despite a highly significant reduction in [(3)H]nicotine binding. An intense signal of beta4 immunoreactivity was found in human dorsal root ganglia, but not in temporal cortex or putamen samples. Synaptophysin immunoreactivities were also similar in Parkinson's disease and control cases. These results suggest that the loss of nicotine binding in the putamen in Parkinson's disease may involve an nAChR subunit (e.g., alpha5 and/or alpha6) other than those investigated. Alternatively, the results could reflect impaired subunit assembly at the plasma membrane. PMID- 11044754 TI - Antagonism of the discriminative and aversive stimulus properties of nicotine in C57BL/6J mice. AB - Mice of the C56BL/6J strain were trained to discriminate between nicotine (1.2 mg/kg) and saline in a two-lever drug discrimination procedure under a tandem variable-interval 60 s fixed-ratio 10 schedule of food reinforcement. Mice of the same strain were trained in conditioned taste aversion (CTA) experiments where drinking a saccharin or saline solution was paired with injection of nicotine or vehicle. During testing with both flavours presented simultaneously, a reduction in the intake of the nicotine-paired solution indicated CTA. The nicotine discrimination was acquired successfully and nicotine yielded a steep dose response curve. The competitive nicotinic antagonist dihydro-beta-erythroidine (DHbetaE, 0.6-3.0 mg/kg) shifted the dose-response for the discriminative stimulus effect of nicotine to the right; the alpha7 nicotinic receptor antagonist methyllycaconitine (MLA, 1.0-10 mg/kg) had no effect. The mice showed strong CTA to 2.0 mg/kg of nicotine and marginally to 0.6 and 1.2 mg/kg of nicotine. DHbetaE (3.0-5.6 mg/kg) attenuated the CTA while MLA (1.0-10 mg/kg) had no effect. These studies show that nicotine has discriminative and aversive stimulus properties in C57BL/6J mice and that the effects are mediated primarily by receptors sensitive to DHbetaE; there was no evidence for the involvement of alpha7 nicotinic receptors. PMID- 11044755 TI - Kappa-opioid receptor modulation of nicotine-induced behaviour. AB - The ability of kappa-opioid receptor ligands to modulate dependence-related behavioural effects of drugs like morphine and cocaine is well documented. The present study examined the effects of kappa-opioid agonists on nicotine-induced locomotor stimulation in rats chronically pre-exposed to nicotine (0.4 mg/kg/day). U50,488 [0.5-3 mg/kg subcutaneously (s.c.)], U69,593 [0.08-0.32 mg/kg intraperitoneally (i.p.)] and CI-977 (0.005-0.02 mg/kg s.c.) administered 30 min prior to nicotine (0.06, 0.2 and 0.4 mg/kg s.c.) dose-dependently antagonised its acute locomotor-activating effect, which was completely prevented by the highest tested dose of each agonist. Baseline activity was unaffected by the largest doses of U50,488 and U69,593, but it was reduced by 0.01 and 0.02 mg/kg of CI 977. The selective kappa-opioid receptor antagonist nor-BNI [30 microg intracerebroventricularly (i.c.v.)] blocked the effects of U69,593 on nicotine induced behaviour, thus supporting the involvement of kappa-opioid receptors in this effect. In conclusion, the activation of kappa-opioid receptors clearly prevented nicotine-induced locomotor stimulation. The effects of at least two of the kappa-opioid agonists were not due to a general motor suppression. It is suggested that the mechanism entails a depression of nicotine-induced increases in accumbal dopamine by these compounds. The results should encourage further research on the role of the kappa-opioid system in the behavioural and neurochemical effects of nicotine, including those related to nicotine dependence. PMID- 11044756 TI - Q&A about this issue PMID- 11044757 TI - Does drug therapy of obesity have a future? PMID- 11044758 TI - Appraisal of drug therapy for nausea and vomiting of pregnancy: I. The placebo effect - methodological and practical considerations. AB - BACKGROUND: Despite that nausea and vomiting of pregnancy (NVP) is the most common medical problem during pregnancy, the placebo effect in the treatment of this condition has not been properly evaluated. METHODS: Two published randomized, controlled trials comparing the antiemetic effects of vitamin B6 with placebo were analyzed. RESULTS: When the severity of nausea or vomiting in the placebo groups at baseline was compared with that at the end of the trials, there was a clear time-dependent placebo effect that peaked between days 4 and 5. The placebo effect appeared to be more pronounced in nausea than in vomiting. CONCLUSIONS: There is a clear placebo effect on NVP. The effect appears to be stronger with nausea than with vomiting. PMID- 11044759 TI - Critical appraisal of drug therapy for nausea and vomiting of pregnancy: II. Efficacy and safety of diclectin (doxylamine-B6). AB - Nausea and vomiting of pregnancy is the most common condition in pregnancy and affects up to 80% of all pregnant women. There are a large number of pharmacological agents that are effective for the treatment of nausea and vomiting associated with conditions such as motion sickness and gastrointestinal conditions; however, their use in pregnancy is limited by the lack of sufficient data on their potential teratogenic effects. The efficacy of the delayed-release combination of doxylamine and pyridoxine (Bendectin, Diclectin) has been shown in several randomized, controlled trials. The present review aims to refute the unsubstantiated beliefs that Diclectin is unsafe when used in the treatment of nausea and vomiting of pregnancy. PMID- 11044760 TI - Efficacy and cost of HMG-CoA reductase inhibitors in the treatment of patients with primary hyperlipidemia. AB - BACKGROUND: Screening for hyperlipidemia is a substantial cost burden, as is its treatment. The choice of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) and the dose level may have significant implications for both efficient and cost effective therapy. OBJECTIVE: To compare the efficiency and cost of statins. MATERIALS AND METHODS: A meta-analysis was conducted of randomized, controlled trials of monotherapy with fixed doses of statins published in the literature until June 1998. Two authors independently extracted data from 49 trials comprising 14,130 patients. The percentage reduction (95% confidence intervals) of low density lipoprotein (LDL) cholesterol levels was calculated using a random-effects model. Cost efficiency was defined as the percentage decline of LDL cholesterol per dollar of drug cost. RESULTS: The population evaluated had a mean baseline LDL cholesterol concentration of 5.31 mmol/L, a mean age of 53.5 years and a mean 59% proportion of males. In reducing LDL cholesterol concentrations to less than 25% of the baseline concentration, a significantly higher cost efficiency was achieved with simvastatin 2.5 mg ( 53.3%/dollar). By targeting a reduction between 25% and 29%, significantly higher cost efficiencies were found with simvastatin 5 mg (-28.9%/dollar), cerivastatin 0.2 mg (-23.8%/dollar) and fluvastatin 40 mg (-23.3%/dollar). For reductions in LDL cholesterol concentrations of 30% to 34%, statistically higher cost efficiencies were achieved with simvastatin 20 mg (-15.0%/dollar) and pravastatin 40 mg (-14. 4%/dollar). Finally, atorvastatin 10 mg yielded a value of -22. 9%/dollar for a 36% reduction in LDL cholesterol concentration. CONCLUSIONS: At current prices of the various doses of statins, the cost efficiency of standard and more aggressive therapies varies substantially. In the context of health care budgets, targeting at-risk patients and using statins judiciously should facilitate the efforts of clinicians and patients to reduce lipid profiles optimally and decrease the cost burden. PMID- 11044761 TI - Rational antipsychotic polypharmacy. AB - The use of two antipsychotics for the treatment of individuals with psychiatric disorders is not uncommon in clinical practice but is rarely documented in the literature. The present article suggests an alternative method of treating individuals who show a partial but inadequate response to antipsychotic monotherapy. A rational strategy for augmenting one antipsychotic with a second, based on pharmacodynamic and pharmacokinetic principles, is provided. The present approach considers the 5-hydroxytryptamine2 to dopamine2 ratio, other complementary receptor affinities and drug-drug interactions involving the cytochrome P450 isoenzymes. The present article presents a rationale for augmenting haloperidol in patients who are partially responsive to clozapine. PMID- 11044762 TI - Reducing substance dependence in elderly people: the side effects program. AB - OBJECTIVES: To reduce benzodiazepine, narcotic and total prescription medication use in community-dwelling elderly people with suspected substance dependence. METHODS: A community-based substance dependence program for seniors was established, and referrals were accepted from a wide variety of sources (self referral, families, emergency departments, family physicians, community agencies, etc). The service typically included several home visits by a nurse and/or social worker trained in recognizing and treating substance dependence in the elderly, medical assessment by a geriatrician and generation of a client-directed treatment plan. Treatment plans included individual and family counselling, recommendations regarding medication changes and involvement in a peer support/education group. Medication use, health care utilization, functional status, cognition and depression scales were collected before and six months after intervention. RESULTS: Of the 95 elderly patients with substance dependence that were seen over a period of 12 months, 55 agreed to participate in the program. Substance dependence included alcohol dependence in 23 participants, benzodiazepine dependence in 20 participants, narcotic dependence in six participants and mixed dependence in six participants. Involvement in the program was associated with significant reductions in depression scale scores (P=0.02), number of daily prescription medications (P=0.002), benzodiazepine use (P=0.01), narcotic use (P=0.04) and number of acute hospitalizations (P=0.001). No significant changes were noted in cognition, functional status, total prescription costs, office visits or emergency room visits. CONCLUSIONS: A community-based sub- stance dependence program for elderly people may significantly reduce narcotic, benzodiazepine and prescription medication use in this population. Further studies are needed to determine the cost effectiveness of such programs. PMID- 11044763 TI - Catch-up growth in 166 small-for- gestational age premature infants weighing less than 1,000 g at birth. AB - Catch-up growth was studied in 166 children born with an extremely low birth weight (<1,000 g) and small-for-gestational age (SGA, <10th percentile birth weight for gestational age). Of these children 159 were followed up for between 4 and 18 years (median 9 years). Group A, SGA <10th percentile of Lubchenco curves only; group B, <10th percentile of Mamelle's curves but >5th percentile, and group C, <5th percentile of Mammelle's curves. Catch-up growth was considered to be achieved when height, weight, and head circumference (HC) reached -2 SD of French reference data and remained above this limit afterwards. Catch-up growth in height was achieved in 126/156 children or 81% (group A 88%; group B 84%; group C 74%), before 3 years of age in 100/127 (78%). Seven children below -2 SD received growth hormone (1 child who caught up was excluded from the results). Catch-up growth in weight was achieved in 126/159 children or 79% (group A 86%; group B 82%; group C 73%), before 3 years of age in 87/126 (69%). Catch-up growth in HC was achieved in 126/156 or 81% (group A 78%; group B 92%; group C 77%), before 3 years of age in 102/127 (80%). Overall catch-up growth was achieved for all three parameters in 65% of children, two of three parameters in 19%, and one of three parameters in 8%. Eleven children never caught up on any parameter. While weight is a lesser concern and HC is not liable to intervention, a greater number of short children might benefit from growth hormone therapy. PMID- 11044764 TI - Duration of home monitoring for infants discharged with apnea of prematurity. AB - Sixty-four preterm infants with apnea of prematurity (AOP) discharged with cardiorespiratory home monitoring (HM) were prospectively followed. For each monitor alarm the parents recorded the occurrence of apnea, bradycardia or color change, and the type of assistance provided. The mean gestational age at birth was 28.8 (26-34) weeks, and the mean birth weight was 1,180 (730-2,390) g. The parents of 61/64 infants (95%) reported a total of 185 true AOPs with a mean of 3 (1-12) events/infant. The mean postconceptional age (PCA) at the last apnea was 41.0 (37-44) weeks. In 80%, AOP terminated between 40 and 44 weeks PCA. There was no correlation between the degree of prematurity and the PCA of the last apnea. We conclude that in preterm infants with AOP discharged with HM, HM may be discontinued at 45 weeks PCA. PMID- 11044765 TI - Spontaneous motility in preterm infants treated with indomethacin. AB - OBJECTIVE: To determine in preterm infants with a patent ductus arteriosus (PDA) the effect of indomethacin treatment on spontaneous motor activity. STUDY DESIGN: Motor activity was assessed from repeated videotape recordings in 32 preterm infants ( 0.96) or O(2) + NO. Matrix metalloproteinase (MMP) activity and MMP-2 mRNA were also measured. In recovery experiments, we measured lung collagen content in piglets exposed to RA + NO or O(2) + NO and then allowed to recover for 3 days. The results show that lung collagen increased 4-fold in the RA + NO piglets, the O(2) and O(2) + NO groups had only a 2-fold elevation relative to RA controls. Unlike the RA + NO piglets, the O(2) and O(2) + NO groups had more than 20-fold elevation in lung lavage fluid hydroxyproline compared to the RA group. O(2) and O(2) + NO also had increased lung MMP activity, extravascular water, and lavage fluid proteins. MMP 2 mRNA levels were unchanged. After 3 days' recovery in room air, the RA + NO groups' lung collagen had declined from 4-fold to 2-fold above the RA group values. The O(2) + NO group did not decline. Alveolar septal width increased significantly only in the O(2) and O(2) + NO groups. We conclude that 5 days' exposure to NO does not result in pulmonary matrix degradation but instead significantly increases lung collagen content. This effect appears potentially reversible. In contrast, hyperoxia exposure with or without NO results in pulmonary matrix degradation and increased lung collagen content. The observation that NO increased lung collagen content represents a new finding and suggests NO could potentially induce pulmonary fibrosis. PMID- 11044770 TI - Effect of polyclonal anti-TNFalpha antibody on endotoxic shock in suckling rats. AB - The aim of this study was to evaluate the effect of anti-tumor necrosis factor alpha (TNFalpha) antibodies on the TNFalpha gene expression in a neonatal septic shock model. Ten-day-old Sprague-Dawley rats were divided into four groups and given intraperitoneal (ip) injection as follows: group 1: 0.1 ml saline; group 2: 0.1 mg/kg Salmonella enteritidis lipopolysaccharide (LPS); group 3: 1 mg/kg of anti-TNFalpha antibodies (Ab); group 4: 0.1 mg/kg of LPS and 1 mg/kg of Ab. We found that in group 2, LPS induced shock, demonstrating hypoglycemia and lactacidemia (p < 0. 05) and death (81.8%). Ab decreased the mortality significantly (35%) and attenuated the hypoglycemia (35 +/- 8 mg/dl in group 2 vs. 53 +/- 3 mg/dl in group 4) and lactacidemia (5.40 +/- 0.63 vs. 2.35 +/- 0.45 mmol) at 8 h in group 4 when compared to group 2. Northern blot demonstrated a significant decrease in TNFalpha mRNA expression in group 4 as compared to group 2, at 2 h after LPS injection. We conclude that the beneficial effects of anti TNFalpha antibodies on LPS-induced shock may be due to decreased TNFalpha gene expression. PMID- 11044771 TI - Plasminogen did not affect lung function in surfactant-treated preterm lambs. AB - Preterm infants with respiratory distress syndrome develop fibrin-rich hyaline membranes within the alveoli and have depressed fibrinolytic activity, which is thought to be due to a relative deficiency of plasminogen. Local fibrin deposition inhibits surfactant function and amplifies inflammation. We hypothesized that plasminogen administration to surfactant-treated preterm lambs would prevent fibrin-rich hyaline membrane formation, resulting in the amelioration of lung pathology and improved lung function. We randomly treated preterm lambs (gestational age 127-129 days) with either 16 mg of lysine plasminogen (n = 10) or saline (n = 10), and ventilated them for 5 h. There were no significant differences in physiologic measurements of lung function (ventilation efficiency index, oxygenation index, dynamic compliance, quasi static pressure volume curve), measures of lung injury (alveolar wash protein content and (125)I-albumin recovery) or surfactant pool size. The degree and extent of bronchiolar erosion and hyaline membrane formation were similar in the two groups. Plasminogen administration did not improve lung function or prevent hyaline membrane formation in surfactant-treated lambs. PMID- 11044772 TI - Adenosine and ATP cause nitric oxide-dependent pulmonary vasodilation in fetal lambs. AB - We investigated the hypothesis that the purine nucleotide ATP and its nucleoside adenosine cause pulmonary vasodilation in fetal lambs by the release of nitric oxide (NO). We also investigated the potential role of K(+)(ATP) channels in mediating the effects of ATP and adenosine on NO. We surgically prepared 28 fetal lambs to measure pulmonary and systemic pressures and pulmonary flow. We investigated the effects of glibenclamide and pinacidil (inhibitor and agonist, respectively, for K(+)(ATP) channels), N-nitro-L-arginine (N-LA) and its methyl ester, N-nitro-L-arginine methyl ester (L-NAME) (inhibitors of endothelium derived NO synthesis), and U46619 (a thromboxane mimetic) on pulmonary vasodilation caused by adenosine and ATP. Adenosine decreased the pulmonary artery pressure and pulmonary vascular resistance (PVR) at doses of 0.08-2.5 microM/kg/min and increased the left pulmonary flow at doses of 0.3-2.5 microM/kg/min in control experiments. N-LA, L-NAME and glibenclamide attenuated the effects of adenosine at doses of < 2.5 microM/ kg/min and pinacidil potentiated its effects. ATP decreased the pulmonary artery pressure and PVR and increased the pulmonary flow at doses of 0.15-2.5 microM/kg/min in control experiments. N-LA and L-NAME attenuated the effects of ATP at doses of < 2.5 microM/kg/min, whereas glibenclamide and pinacidil had no effect on the response to ATP. U46619 increased the basal pulmonary vascular tone, but did not significantly alter the vasodilative responses to ATP and adenosine. In conclusion, adenosine and ATP cause NO-dependent pulmonary vasodilation in fetal lambs. The activation of K(+)(ATP) channels plays a role in adenosine-induced pulmonary vasodilation. The mechanism by which ATP causes NO release and pulmonary vasodilation requires further investigation. PMID- 11044774 TI - Chronic lung disease-cracking the condition in the 21st century PMID- 11044773 TI - Plasma carbon monoxide levels in term newborn infants with sepsis. AB - Carbon monoxide (CO) has been implicated as a new endogenously produced mediator similar to nitric oxide (NO). CO was measured in plasma samples from 7 term newborn infants with sepsis and from 30 healthy neonates. Plasma CO levels were significantly higher in the group with sepsis at the time of admission to the neonatal intensive care unit than in the healthy controls (p < 0.05). Moreover, the elevated plasma CO levels were significantly related to increased NO production, as indicated by plasma nitrite/nitrate levels (p < 0.05). The present study suggests that, in addition to NO, CO might be another important mediator taking part in the pathogenesis of neonatal sepsis. PMID- 11044775 TI - Clinical efficacy and safety of donepezil on cognitive and global function in patients with Alzheimer's disease. A 24-week, multicenter, double-blind, placebo controlled study in Japan. E2020 Study Group. AB - This study evaluated efficacy and safety of donepezil hydrochloride (donepezil) at 5 mg/day in patients with mild to moderately severe Alzheimer's disease for 24 weeks in a double-blind, placebo-controlled comparative trial. In this study, 268 patients were enrolled and 39 of these (15%) were withdrawn. In the evaluable population of efficacy, Protocol-Compatible (PC) analyzed patients (n = 228), better effects than that of placebo were confirmed using two primary efficacy measures: a cognitive performance test, the Japanese version of the Alzheimer's Disease Assessment Scale - cognitive subscale (ADAS-J cog, p = 0.003) and a clinical global assessment, the Japanese version of the Clinical Global Impression of Change (J-CGIC, p = 0.000). The superiority of donepezil was also shown by secondary measures: the Sum of the Boxes of the Clinical Dementia Rating (CDR-SB), the Mental Function Impairment Scale (MENFIS) and the caregiver-rated modified Crichton scale (CMCS). The same results were obtained in the intention to-treat (ITT) analysis (n = 263). The incidence of drug-related adverse events was 10% (14/136) in the donepezil and 8% (10/131) in the placebo group; no significant difference was seen between the two groups. The main adverse events were gastrointestinal symptoms, and these were almost all mild, and they all disappeared with continued administration or temporary discontinuation of donepezil. These results indicate that the donepezil appears to be effective and well tolerated in patients with mild to moderately severe Alzheimer's disease. PMID- 11044776 TI - Acetylcholinesterase inhibition in dementia with Lewy bodies: results of a prospective pilot trial. AB - OBJECTIVE: Dementia with Lewy bodies (DLB) is the second commonest form of dementia. The response to acetylcholinesterase inhibition (AChEI) could be greater in DLB than in Alzheimer's disease (AD) because cholineacetyl-transferase levels are more reduced in the former. This preliminary trial seeks to compare performances in cognitive tasks before and after tacrine administration in DLB and AD subjects. METHODS: Six DLB and 6 AD patients were enrolled in an open, nonrandomized, intervention trial using 80 mg/day tacrine. Patients met ADRDA or DLB consortium criteria for probable diseases. Subjects were matched for Mini Mental State Examination (MMSE) score, age and sex. Mattis Dementia Rating Scale (DRS), Controlled Oral Word Association Test (FAS) and Boston Naming tests were administered at baseline and at 6 months into treatment. RESULTS: AD and DLB groups did not differ in initial mean total DRS scores. In the primary analysis, both groups declined during the course of treatment (-7.3 +/- 4.2 and -16.8 +/- 39.2 DRS points, respectively). Due to the large variability in DLB posttreatment scores, this group was divided post hoc into responders (DLBr) and nonresponders (DLBnr). The DLBr group outperformed the DLBnr group at baseline (p < 0.05) and, notably, in follow-up DRS test scores (p < 0.001). Two-way MANOVA comparing both DLB subgroups with either the entire AD cohort or similarly stratified AD subgroups showed a significant interaction (F = 7.6; p < 0.015), attributed mostly to declines in DLBnr group scores (p < 0.01). Surprisingly, on DRS memory subscale and FAS tests, there were significant improvements in DLBr scores (p < 0.02). A baseline MMSE (or DRS memory) score >/=15 predicted a positive response to tacrine in DLB. Acceleration of parkinsonism occurred in all DLB subjects. CONCLUSION: Results from a primary analysis of the therapeutic effect of 80 mg/day tacrine in DLB and AD were negative. However, post hoc analysis showed that mild to moderate DLB responds favorably to AChEI relative to AD through stabilization of global cognitive decline and improvements in specific cognitive areas. These results could be useful in the planning of a more definitive study. PMID- 11044777 TI - Intrathecal release of nitric oxide in Alzheimer's disease and vascular dementia. AB - A growing body of evidence points out the potential role of inflammatory mechanisms in the pathophysiology of brain damage in dementia. We have recently demonstrated that patients with Alzheimer's disease (AD) and vascular dementia (VaD) display an intrathecal production of proinflammatory cytokines. TNF-alpha, one of these cytokines, leads to the production of nitric oxide (NO), a potent inflammatory mediator, by induction of inducible NO synthase. The aim of the present study was to investigate the intrathecal levels of nitrate, one of the main metabolites of NO, and to relate its levels to the degree of intellectual impairment, in patients with AD and VaD. Twenty patients with early AD and 26 patients with VaD were analyzed with respect to cerebrospinal fluid levels of nitrate by gas chromatography/mass spectrometry. Interestingly, in patients with AD but not VaD, the intrathecal levels of nitrate were significantly and inversely correlated (r = -0.68, p = 0.002) to the degree of intellectual impairment. Our study demonstrates an inverse correlation between the intrathecal levels of nitrate and the degree of cognitive impairment in patients with AD, suggesting a neuroprotective effect of NO in AD. PMID- 11044778 TI - Quantification and characterization of fluctuating cognition in dementia with Lewy bodies and Alzheimer's disease. AB - Fluctuating cognition (FC) is a common and important symptom in dementia, particularly dementia with Lewy bodies (DLB), although it has not been empirically quantified or characterised. Forty subjects (15 DLB, 15 AD, 10 elderly controls) were evaluated using a clinical FC severity scale, as well as receiving measures of variability in attentional performance and slow EEG rhythms across 90 s, 1 h and 1 week. DLB patients had significantly more severe FC and more severe variability in attentional and slow electrocortical measures than either AD patients or normal controls in all time frames. Attentional and EEG variability also correlated significantly with independent clinical ratings of FC. Clinical quantification and measures of attention and EEG variability can therefore make an important and standardised contribution to the assessment of FC in dementia, facilitating future treatment studies with important implications for the potential causative mechanisms and differential diagnosis. PMID- 11044779 TI - Impaired processing of famous faces in Alzheimer's disease is related to neurofibrillary tangle densities in the prefrontal and anterior cingulate cortex. AB - To examine the neuroanatomical correlates of impaired processing of famous faces in Alzheimer's disease (AD), we performed an anterograde clinicopathological study of 25 patients with clinically and neuropathologically confirmed AD. Famous face recognition, identification and naming was assessed using the Famous Face Test. The assessment of neurofibrillary tangle (NFT) and senile plaque (SP) densities was performed in ten cortical areas in both hemispheres, and statistical analysis was made using forward stepwise logistic regression models. A statistically significant relationship was found between NFT densities in Brodmann's areas 9 and 24 in both hemispheres and impaired famous face naming and identification. SP counts did not correlate with any of the neuropsychological parameters. These data suggest that NFT formation in prefrontal and anterior cingulate cortex, two areas involved in semantic memory processes, is a key event in famous face naming and identification deficits. In agreement with previous studies, they also indicate that SP densities are not a good pathological correlate of neuropsychological deficits in AD. PMID- 11044780 TI - Fate of patients with questionable (very mild) Alzheimer's disease: longitudinal profiles of individual subjects' decline. AB - Currently used criteria for the diagnosis of probable Alzheimer's disease requires the presence of cognitive deficit in addition to loss of episodic memory. The recent developments in drug treatments for Alzheimer's disease have highlighted the importance of early diagnosis and the need to characterise the cognitive profile of the earliest stages of the disease. We set out to examine the pattern of decline in terms of individual cognitive domains in non-demented subjects with clinically isolated progressive amnesia which we have termed questionable Alzheimer's disease. Twelve subjects who fulfill criteria for a clinical diagnosis of possible Alzheimer's disease by NINCDS-ADRDA criteria were compared to 20 age-matched controls in a longitudinal study. All subjects had MMSE scores of 24 or greater at entry to study. Individual profiles were measured by impairment in the cognitive domains of episodic memory, attention, semantic memory, visuospatial function and auditory verbal short-term (working) memory. Even after subjects were given this intensive neuropsychological battery, 8 of 12 subjects had episodic memory deficits only. Within 12 months just 3 patients were only amnesic and the other patients with pure amnesia at year 1 developed either semantic or attentional deficits. Impairment in both these cognitive domains preceded impairment in visuospatial function and auditory-verbal short-term memory. Our findings are consistent with the pattern of cognitive impairment in Alzheimer's disease increasing in accordance with neuropathological correlates of cognitive function with amnesia linked to the initial medial temporal pathology before the pattern of spread affects critical neural substrates for attention and semantic memory. Tests of selective attention and semantic memory appear to be the most sensitive markers of decline beyond the amnestic phase of early Alzheimer's disease. PMID- 11044781 TI - People with dementia use schemata to support episodic memory. AB - This paper describes two experiments which examine whether people with dementia use schemata, established knowledge stores associated with specific scenarios, to aid episodic memory. On all tests of memory, volunteers with dementia remembered less than their age-matched controls. There was evidence for an age-related decline in cognitive resources available for effortful memorizing. For people with dementia, errors of commission in free recall, false recognition scores and reconstruction data were consistent with the use of schema to guide retrieval of items from memory. We conclude that retrieval support improves memory performance for both the healthy and dementing elderly. More importantly, pre-established schemata remain intact well into the course of dementia and can support recall of newly acquired episodic information. PMID- 11044782 TI - Initial lesions of vascular aging disease (arteriosclerosis). AB - BACKGROUND: In contrast to the well-known morphologic lesions of arteriosclerosis, the initial changes of the disease are less obvious. Commonly, functional disturbances of the endothelium, endothelial dysfunction, are suggested. On the other hand the significance of age-dependent changes in the extracellular matrix with their important role in vessel wall permeability and other features associated with arteriosclerosis should not be overlooked. New topics deal with possible infectious factors, the genetic basis of the disease and the particularities of the unstable atheroma. OBJECTIVE: Alterations in nitric monoxide and endothelin-1 balance of the endothelium are the key events in the initiation of arteriosclerosis induced by oxidized lipoproteins, cigarette smoking and endotoxin. This frequently supposed mechanism contrasts with earlier opinions on the primary alterations in glycosaminoglycan metabolism and other components of the extracellular matrix against atherogenic factors like hypertension, stress and physical inactivity. Based on a survey of the literature and our own experimental experiences, these changes in connection with the morphometrically determined age-conditioned increase in vascular wall thickness and the above-mentioned new topics on arteriosclerosis were analyzed. CONCLUSION: The initial lesions of arteriosclerosis starting in youth seem to be fundamentally different from those beginning in old age. The first step in the development of fatty streaks in the arteries of young people is endothelial dysfunction with a decreased formation of nitric monoxide and an increased expression of adhesion molecules. In comparison the genesis of arteriosclerosis in advanced age is characterized by metabolic changes in the endothelium combined with age-conditioned alterations in the extracellular matrix resulting in faster progression of the disease in old age. The multicausal genesis of arteriosclerosis cannot be doubted even if cooperation with infectious factors cannot be excluded. The histopathologic peculiarities of unstable atheroma are described. PMID- 11044783 TI - Aging and spatial localization during feature search. AB - BACKGROUND: Previous studies indicate that older adults, like young adults, can efficiently search for a briefly presented visual target defined by a single salient feature presented amidst background distractors. However, little is known about older adults' ability to identify the spatial location of targets during this aspect of preattentive processing. OBJECTIVE: This study examined the extent to which older adults exhibit localization problems during feature search for a target with high conspicuity. Their performance was compared to that of younger adults. METHODS: Twenty older adults (mean age 70 years, 8 men and 12 women) and 20 younger adults (mean age 25 years, 6 men and 14 women) with good central and peripheral vision were tested. Subjects were asked to indicate via a computerized touch-screen the location of a briefly presented (80 ms) target presented amidst distracting stimuli (set size 8, 16, or 32). Targets were presented at either 10 degrees, 20 degrees, or 30 degrees eccentricity. The dependent measures were percent correct localization and, for trials in which there were errors, the spatial magnitude of the error. RESULT: Compared to young adults, older adults committed more localization errors during feature search, a problem which was accentuated with increasing target eccentricity. In addition, older adults' mislocalizations deviated from the correct location by greater distances. CONCLUSIONS: Older adults have spatial localization problems in preattentive processing during feature search, which could be detrimental to the guidance and deployment of visual attention. PMID- 11044784 TI - Visual contributions to postural stability in older adults. AB - BACKGROUND: With advancing age, there is a generalized reduction in visual functioning which has been associated with impaired postural stability and increased risk of falls. However, little is known about which visual abilities are the most important in the control of postural sway when standing. OBJECTIVE: To determine whether specific visual abilities predict stability when standing on firm and compliant surfaces. METHODS: Tests of visual function, peripheral sensation, strength, reaction time and sway were administered to 156 community dwelling men and women aged 63-90 years. The visual tests included high- and low contrast visual acuity, contrast sensitivity, depth perception, stereopsis and lower visual field size. Postural sway was measured with eyes open on a firm and a compliant foam rubber surface. RESULTS: On the firm surface, sway was significantly associated with only one sensorimotor measure: proprioception in the lower limbs. In contrast, on the compliant surface, sway was associated with all of the visual measures, quadriceps strength and reaction time. Multiple regression analysis revealed that contrast sensitivity, stereopsis and quadriceps strength were significant independent predictors of total sway when subjects stood on the compliant surface. CONCLUSION: The study findings confirm the importance of vision, in particular contrast sensitivity and stereopsis, in the control of posture under challenging conditions, and suggest some mechanisms for the association between impaired vision and falls in older people. PMID- 11044785 TI - The effect of anti-platelet aggregation to prevent pressure ulcer development: a retrospective study of 132 elderly patients. AB - BACKGROUND: As the number of bedridden elderly patients increases, prevention of pressure ulcers is becoming a more important issue. However, an approach to this problem using medication has not been considered sufficiently in the clinical context. OBJECTIVE: To test the hypothesis that anti-platelet aggregation therapy administered to the elderly patients may be helpful in preventing pressure ulcer formation, the medical records of 132 bedridden elderly patients were analyzed. In addition, the propensity of platelets to aggregate was also measured in some of the bedridden patients. METHODS: Patients were divided into two groups, with pressure ulcers (group P, 52 patients) and without (group N, 80 patients). Subsequently, six factors defining the clinical characteristics age and gender, underlying disease, cause of being bedridden, level of consciousness, mobility and activity as defined on the Braden scale, and frequency of anti-platelet aggregation medication were investigated in groups P and N. In addition, physical findings (three factors): body mass index, blood pressure (BP), and heart rate were investigated in both groups. Furthermore, laboratory data (seven factors): total protein (TP), albumin, total cholesterol, hemoglobin (Hb), hematocrit (Hct), platelets, and platelet aggregation were compared between two groups. RESULTS: There were no significant differences in clinical characteristics between the two groups with the exception of the frequency of anti-platelet aggregation medication (23.1% of group P vs. 40.0% of group N, chi(2) = 4.06, p < 0.05). There was also no significant difference in physical findings except a difference between systolic and diastolic BP (48.4 mm Hg in group P vs. 57.1 mm Hg in group N, p < 0.01). Values of TP, albumin, Hb, and Hct in both groups were lower than the normal range, but there was no significant difference between the two groups. The platelet count was significantly greater in group P than in group N after lying supine (p < 0.03), and platelet aggregation in group P was significantly higher compared with group N (p < 0.03). CONCLUSIONS: Although our hypothesis must be tested by a randomized prospective trial, these results indicate that increased platelet aggregation is possibly associated with the development of pressure ulcers; therefore anti-platelet aggregation therapy may prevent their occurrence in bedridden elderly patients. PMID- 11044786 TI - Sleep patterns and mortality among elderly patients in a geriatric hospital. AB - BACKGROUND: Sleep disturbance is one of the major and unsolved problems in older people. Most of the previous sleep studies rely on self-reported documents, and memory disturbance in older people might bias sleep complaints and health status. OBJECTIVE: Sleep disturbances were studied as a mortality risk. METHODS: In 272 patients who were aged, infirmed and chronically institutionalized in a skilled care geriatric hospital, the presence or absence of sleep disturbances were examined by hourly observations of patients over 2 weeks at baseline, and they were prospectively followed up for 2 years to assess mortality. RESULTS: Mortality after 2 years was significantly higher in the nighttime insomnia, daytime sleepiness, and sleep-onset delay groups. Further, adjusted for age, gender and activities of daily living status, the presence of nighttime insomnia and sleep-onset delay remained associated with a higher risk of mortality. CONCLUSION: Sleep disturbance may be one of the symptoms indicating poor health or functional deficits, and be an independent risk factor for survival. PMID- 11044787 TI - Prevalence and survival of myelodysplastic syndrome of the refractory anemia type in hospitalized cognitively different geriatric patients. AB - BACKGROUND: Myelodysplastic syndrome (MDS) is predominantly a disease of old age. The number of MDS cases diagnosed over the last 20 years has risen substantially due to increased awareness and improved geriatric care. Although MDS is increasingly diagnosed, the prevalence and prognosis of early-stage affected elderly are not completely known. OBJECTIVE: To evaluate the prevalence, characteristics and prognosis of newly diagnosed MDS patients hospitalized in an acute and subacute geriatric department. METHODS: Between 1993 and 1996, 3,275 patients hospitalized in the geriatric department of a teaching hospital for acute care or short-term rehabilitation were investigated for unexplained hematological abnormalities. Demographic, chronic comorbidities, cause of hospitalization, functional and cognitive status, hematological and other laboratory parameters were collected. RESULTS: Two hundred and forty-five (7.5%) patients had unexplained cytopenia, macrocytosis or monocytosis, of whom 37 (15%) were diagnosed as having MDS. Only 9 patients were hospitalized for evaluation of anemia, 28 for infections, cardiac, cerebrovascular events and other causes. Thirty-four patients had refractory anemia (RA), two had RA with ringed sideroblasts and 1 had RA with an excess of blasts (RAEB). The follow-up period was up to 70 months. No differences were found between demented and cognitively normal patients in age, sex, comorbidities or laboratory parameters. Comparison of survival curves (excluding the RAEB case) according to demographic, clinical and hematological parameters has shown that only dementia adversely affects survival, compared to cognitively normal patients (p = 0.024). CONCLUSIONS: MDS of the RA type is a common and incidental finding in older hospitalized patients. It is a frequent cause of anemia and other hematological abnormalities but has less significance on survival rates than dementia, although its full impact remains to be determined. PMID- 11044788 TI - Aging and carcinogenesis--insufficient metabolic cell repair as the common link. AB - BACKGROUND: The mechanisms of the development of cancer in old age and also the mechanisms of aging are not well understood. This paper tries to interpret consequences of malignant tissue transformation from the viewpoint of aging, or in other words, from an insufficient cell adaptation to the needs of repair and proliferation. SUBJECT: A hypothesis is presented that a unified but quite opposite at different stages of ontogenesis mechanism is the basis of atypical growth and embryonic development. In the beginning of a malignant dedifferentiation is an insufficiency of an effective self-renovation and disturbed preservation of its adaptation capability. The suppression of regenerating cell proliferation is the primary event of the development of a dedifferentiated tissue growth. The transformation of normal cells into tumor cells is an adaptive reaction in reply to a shortage of self-regeneration capability and repair. Allowing for the process of rebirth, i.e. the complete restoration of tissues leading to the restrain of senescence proceeds by the type of embryonic growth of tissues, the possibility to use the potential of transformed cells for restraining senescence is proposed. The latter will permit to direct the process of transformation to an integrated growth channel, to prevent the clinical phenomenon of malignancy, and use the potential of transformed cells for realization of the self-renovation program and program of unlimited life duration of the whole organism. CONCLUSION: By a stimulation or compensation of the age-induced shortage of cell metabolism, two effects can be expected: prevention of cancer and retardation of aging. PMID- 11044789 TI - Physical fitness related to disability in older persons. AB - BACKGROUND: In today's aging society, preventing or reducing disability is important. Physical activity may serve this goal. Generally, physical activity aims to enhance physical fitness, which in turn may prevent disability. The relationship between physical fitness and disability has been much less explored than the relationship between physical activity and physical fitness. OBJECTIVE: To investigate the relationship between separate components of physical fitness and disability. METHODS: The subjects were a community-based sample of 176 men and 233 women aged 65 years or older. Physical fitness was assessed with performance-based tests. Disability and potential confounders were assessed during face-to-face interviews. RESULTS: Independently of other fitness components, walking endurance, grip strength, manual dexterity and balance contributed significantly to the prediction of disability for both men and women. Flexibility of the hip and spine, flexibility of the shoulder and reaction time were not independent predictors of disability for men or women. Physical fitness explained a greater percentage of variance in disability for women (31-48%) than for men (14-34%). Although depressive symptoms, cognitive functioning (men), number of chronic conditions (women) and age (women) explained additional variance in disability, these variables did not confound the relationship between physical fitness and disability. CONCLUSION: Walking endurance, grip strength and manual dexterity are important unique predictors of disability. Physical activity programs should be directed at these fitness components. PMID- 11044790 TI - Human sex differentiation: from transcription factors to gender. AB - Over the past decade, knowledge of the genetic control of human sex differentiation has greatly expanded our understanding of the developmental processes needed to form a male or female. The purpose of this review is to discuss how transcription factors are relevant to such processes. Additionally, an attempt is made to relate current knowledge of these factors with gender development of subjects with intersex conditions. Finally, we discuss how information about the genetic control of sex differentiation may contribute to decisions about medical treatment of individuals with conditions of abnormal sex differentiation. PMID- 11044791 TI - Usefulness of HMG-CoA reductase inhibitor in Japanese hyperlipidemic women within seven years of menopause. AB - OBJECTIVE: To assess the therapeutic value of treatment with an HMG-CoA reductase inhibitor in women with hypoestrogenic hyperlipidemia caused by menopause. DESIGN: Fifty-six women with total cholesterol (TC) levels of 220 mg/dl or more who were within 7 years of menopause were randomly assigned to receive an HMG-CoA reductase inhibitor (pravastatin 10 mg/day; treated group, 26 patients) or no medical treatment (nontreated group, 30 patients) in this 6-month nonblinded prospective trial. RESULTS: In the treated group, the mean (SD) TC levels decreased significantly from 254.5+/-22.3 mg/dl at baseline to 204.7+/-22.2 mg/dl (19.6%), and the mean low-density lipoprotein cholesterol (LDL-C) level decreased significantly from 146.7+/-30.5 to 104.3+/-22.5 mg/dl (28.9%); the mean arteriosclerotic index decreased significantly from 2.98 to 2.08 (30.2%). There were no significant changes in either triglyceride levels or high-density lipoprotein cholesterol (HDL-C) levels. In the nontreated group, there were no significant changes in the TC, HDL-C, LDL-C, or triglyceride levels; there was also no change in the arteriosclerotic index. After 6 months, the TC level, LDL-C level, and arteriosclerotic index were significantly lower in the treated group compared with the nontreated group (p<0.01). CONCLUSIONS: The results indicate that the HMG-CoA reductase inhibitor lowered TC and LDL-C levels and was useful in the treatment of hypoestrogenic hyperlipidemia for periods of at least 6 months. PMID- 11044792 TI - Evidence for de novo cholesterol synthesis by term human fetal amnion and chorion: a comparative study using the reverse-isotope dilution technique. AB - The cholesterol biosynthetic activity was assessed using [2-(14)C]-acetate as substrate in the homogenates of amnion and chorion obtained from women (n = 6, age 26-39 years) after spontaneous labour at term (37-40 weeks of gestation) having uncomplicated pregnancies. Reverse-isotope dilution analysis gave positive identification of [(14)C]-cholesterol acetate in all incubations of viable tissues. This metabolite was not evident in heat-denatured homogenates which served as controls. The extent of enzymic conversion for amnion at 2.6 x 10(-3) to 0.19% was persistently higher than that of the chorion at 1.7 x 10(-3) to 9.0 x 10(-3)%. The results indicate that human term fetal membranes possess the full complement of enzymes to catalyze the transformation of acetate to cholesterol. This study provides evidence that fetal membranes possess the capacity for de novo cholesterol biosynthesis, the sterol being essential for steroidogenesis as well as in embryo viability during pregnancy. PMID- 11044793 TI - Perceived versus measured height. Which is the stronger predictor of psychosocial functioning? AB - The relationship between perceptions versus measured height and children's psychosocial adaptation in a sample of medically referred youth with short stature was investigated. All children referred for a growth evaluation to one regional pediatric endocrinology clinic received a psychosocial screening assessment as a routine component of their initial visit. Data were collected for patients ages 4-18 years (n = 620) with heights ranging from -4.0 to -1.1 SD for age- and gender-adjusted population norms. Patients (8 years and older) and in all cases a parent/guardian served as informant through paper-and-pencil questionnaires. Both children and parents overestimated the child's height. Overestimations of height were associated with greater patient and parent satisfaction with stature. Perceived height was more strongly associated with psychosocial adaptation than was measured height. Clinical management decisions designed to enhance patient quality of life by increasing projected adult height through hormonal interventions should take into account both measured and perceived patient height. PMID- 11044794 TI - Catch-up growth in short-at-birth NICU graduates. AB - The statural catch-up growth, defined as reaching at least tenth length/height percentile (P10) for normal population standards (-1.28 SD score, SDS), was studied in 73 infants short at birth (length < P10 for gestational age) admitted to NICU. Mean gestational age at birth was 35.2 weeks (range 29-41) and mean birth length standard deviation score -2.31 (-4.52/-1.46). Infants were measured at birth, at 3, 6, 12, 18, and 24 months corrected age and then once a year until 6 years chronological age. Statural catch-up growth was studied, with reference both to normal population standards and to individual genetic target. With reference to normal population standards, 44% of infants had caught-up at 3 months of age, 51% at 3 years, 66% at 4 years and 73% at 6 years. In the case of individual genetic targets, a similar trend was present, but the absolute values were slightly higher from 4 to 6 years (73 vs. 66% and 78 vs. 73%, respectively). Statistically significant changes in mean standard deviations score for chronological age were present from birth to 3 months, 3 to 12 months, 3 to 4 years and 5 to 6 years (p<0.05). No differences were found in this trend of recovery when considering ponderal index (PI) at birth (symmetrical vs. asymmetrical), sex (male vs. female) or gestational age (p>0.05). In the majority of cases infants with short stature at birth admitted to a NICU had a statural catch-up growth within the first years of life. This is more evident when considered in relation to individual genetic target rather than to normal population standards. PMID- 11044795 TI - Estrogen receptors and the activity of nitric oxide synthase in the artery of female rats receiving hormone replacement therapy. AB - OBJECTIVE: To observe the expression of estrogen receptor and the activity of NOS in the arteries of female rats receiving estrogen replacement therapy. METHODS: Seventy-two female rats were randomly divided into four groups: group A: sham ovariectomy; group B: ovariectomy; group C: ovariectomy with estrogen replacement therapy (benzoate estradiol, 5 microg i.m. once in 2 days); group D: ovariectomy with estrogen and progesterone replacement therapy (benzoate estradiol, 5 microg i.m. once in 2 days and progesterone, 1 mg i.m. once in 2 days). The rats were killed after 2 months. The receptor-binding assay was adopted to measure the estrogen receptors in the arteries of the rats, and the activity of NOS in the arteries was assessed by the hemoglobin reductase method. RESULTS: The ER number and NOS activity in the arteries of the ovariectomized group are less than those in sham-ovariectomy group (p<0.05). The ER number and NOS activity in the arteries of groups C and D are larger and higher than those in the ovariectomized group (p<0.05). No significant differences in the ER number and NOS activity were observed between groups C and D. CONCLUSION: The ER number and NOS activity in the rat artery significantly decrease after ovariectomy, while hormone replacement therapy can significantly increase the artery NOS activity and retain the ER number in the artery of the ovariectomized rats to normal level. The result may contribute to explaining the beneficial effect of estrogen in the prevention of coronary artery diseases in postmenopausal women. PMID- 11044796 TI - Normative data for total and free acid-labile subunit of the human insulin-like growth factor-binding protein complex in pre- and full-term newborns and healthy boys and girls throughout postnatal development. AB - Insulin-like growth factors (IGFs) circulate in plasma as part of a 150-kD complex that also contains IGF-binding protein-3 (IGFBP-3), a protein that binds IGF-I and IGF-II with high affinity, and an acid-labile subunit (ALS) that does not directly bind IGFs. Because the ALS assay methods currently being used are relatively new, there is a need for updated normative reference data. We report the normative data in 17 preterm infants (10 males and 7 females), 30 normal full term newborns (15 males and 15 females) and 150 normal children who where divided into 5 groups according to their Tanner stage (15 males and 15 females per group). Serum levels of total and free ALS were significantly lower in premature infants than in full-term newborns, but all newborns had significantly lower levels than Tanner stage-I children (p<0.001, ANOVA). A significant increase was seen between Tanner stages I-III in both sexes (p<0.001, ANOVA). No differences were found between sexes at any developmental age studied. Significant correlations (p<0.001) were seen between total and free ALS concentrations and IGF-I (r = 0.50 and 0.60, respectively), free IGF-I (r = 0.37 and 0.36), IGF-II (r = 0.37 and 0.27), IGFBP-1 (r = -0.48 and -0.49), IGFBP-2 (r = -0.44 and -0.51) and IGFBP-3 (r = 0.67 and 0.59) at all Tanner stages. However, no correlation was found with IGFBP-1, -2 or -3 levels at birth. This study shows normal values in a population of preterm infants and healthy Spanish newborns and subjects of both sexes at all stages of pubertal development and indicate the different relationships between the components of the IGF system during intra- and extrauterine life. PMID- 11044797 TI - Case of XXXXY syndrome. Development throughout adolescence and endocrine aspects. AB - 49,XXXXY syndrome is a very rare condition that is associated with a considerable more severe phenotype than classic 47,XXY Klinefelter syndrome. We present a patient with 49,XXXXY syndrome, who was first presented to an endocrinological unit at the age of 12.5 years with prepubertal genitalia. He was then lost from follow-up and showed clear clinical and biochemical signs of hypergonadotropic hypogonadism when presenting again at the age of 16 years. The patient was started on testosterone replacement therapy. This case is reported to underline the need for thorough endocrinological follow-up examinations in males with X polysomies. PMID- 11044798 TI - Effects of octreotide infusion, surgery and estrogen on suppression of height increase and 20K growth hormone ratio in a girl with gigantism due to a growth hormone-secreting macroadenoma. AB - We treated an extremely tall 13-year-old girl with a growth hormone (GH) secreting macroadenoma and GH levels of 120-495 ng/ml with a combination of preoperative octreotide infusion, surgery and postoperative octreotide infusion plus estrogen, which resulted in reduced tumor size prior to surgery, reduced GH levels and completely suppressed growth after surgery. 20K GH is produced by alternative splicing of 22K GH mRNA and the ratio of 20K GH to 22K GH is within a small range in the normal population and high in a GH-secreting tumor. The 20K/22K GH ratio in this patient was persistently elevated during each phase of the treatment and may serve as a sensitive index of tumor-derived GH secretion. PMID- 11044799 TI - Hormonal factors influencing weight and growth pattern in craniopharyngioma. AB - Patients operated on for craniopharyngioma frequently suffer from hyperphagia and are obese, but their statural growth is normal despite growth hormone (GH) deficiency. We have evaluated the hormonal factors influencing changes in weight and growth in 17 children before and 1, 3-6, 12, and/or 24 months after surgical resection of a craniopharyngioma performed at 7.7 +/- (SE) 1 years of age. Of these, 15 patients had a GH deficiency before surgery, and all had complete pituitary deficiency after it. The plasma fasting insulin concentrations before surgery were positively correlated with body mass index (BMI, kg/m(2); p < 0.05), plasma insulin-like growth factors (IGFI, p = 0.03, and IGFII, p = 0.04), and leptin (p = 0.03). They increased significantly 1 month after surgery and continued to increase thereafter, whereas leptin increased significantly only 3-6 months after surgery, paralleling changes in BMI. The plasma fasting insulin concentrations before surgery were also positively correlated with the weight changes (12.3 +/- 2.3 kg, p < 0.01) during the 12 months after surgery, but not with changes in BMI SDS (3.1 +/- 0.5, p = 0.07). Both expressions of weight change were correlated with the concomitant growth rates (4.8 +/- 0.7 cm, p < 0.01). IGFI was above the 10th percentile for children with idiopathic short stature in 10 of 15 patients with craniopharyngioma-induced GH deficiency and IGF binding protein 3 in 14 of 15 patients. Craniopharyngioma itself modified the control of insulin secretion, and surgery increased the insulin secretion which continued in the same way in a given patient after surgery. The increased insulin secretion in turn increases weight and keeps IGFI nearly normal. This may explain the normal growth rate despite the complete lack of GH. PMID- 11044800 TI - Antenatal and early postnatal dexamethasone treatment decreases cortisol secretion in preterm infants. AB - Glucocorticoids are used antenatally to accelerate the maturation of fetal respiratory and cardiovascular systems when a threat of preterm delivery exists. Postnatally, they are used to prevent and treat respiratory distress syndrome. This study investigates the effects of antenatal (ACT) and early postnatal corticosteroid treatment (PCT) on serum cortisol and plasma catecholamine and adenosine 3',5'-cyclic monophosphate (cAMP) concentrations in preterm neonates. The infants in the ACT group had a significantly lower cortisol concentration than the infants in the non-ACT group on the first day of life. After birth, the infants were further divided into non-PCT and PCT groups. PCT suppressed cortisol levels significantly after 2 days, and the cortisol levels were still lower 2 days after discontinuation of PCT. No effect of PCT on plasma cAMP or catecholamine concentrations was observed. The results indicate that both ACT and a short PCT can significantly suppress basal cortisol levels in preterm infants. PMID- 11044801 TI - Biological effects of newly synthesized cholecystokinin analogs. AB - Cholecystokinin (CCK) is a gut hormone that regulates pancreatic endocrine functions via CCK(A) receptors. CCK(4) (Trp-Met-Asp-Phe-NH(2)) has an insulinotropic effect, but is 1,000-fold less potent than CCK(8) in rodents. The in vitro potencies with respect to binding, the biological effects and the selectivity of newly synthesized CCK(4) analogs constructed by computer modelling experiments were investigated in vitro in rat pancreas and brain, INS-1 cells, and guinea pig ileum. Exchanging various amino acids, e.g. Met by either Pro or Nle, and modifying Phe by adding various substituents in different positions led to compounds which were more effective as insulin secretagogues than CCK(4) itself and even show insulinotropic effects comparable with those of CCK(8) (e. g. compounds M1 and M2 being substituted at Phe). Some compounds which possess electron withdrawing groups on the C-terminal Phe and possess a Pro instead of a Met were especially effective. The CCK(A) receptor antagonist L-364,718, but not by the CCK(B) receptor antagonist L-365,260, inhibited the insulinotropic effects. The synthetic CCK(4) compounds were not selective for the endocrine pancreas: e.g. M1 and M2 had binding activity with respect to rat brain homogenates but no activity with respect to contraction of the guinea pig ileum. The data indicate that some of the newly synthesized CCK tetrapeptides exhibit a high affinity for the CCK receptor of beta-cells and have an insulinotropic effect much higher than CCK(4). PMID- 11044802 TI - Growth hormone normalises height, prediction of final height and hand length in children with Prader-Willi syndrome after 4 years of therapy. AB - BACKGROUND: Based on the reported favourable effects of growth hormone (GH) treatment on growth and body composition in Prader-Labhart-Willi syndrome, we studied age dependency and the long-term effects on growth dynamics to elucidate the assumed hypothalamic GH deficiency. METHODS: We examined 23 children treated with hGH (24 U/m(2)/week) during a median of 4 (range 1.5-5.5) years; group 1: 10 young underweight (age 0.3-4.1 years), group 2: 8 prepubertal overweight (age 3.7 9.5 years) and group 3: 5 pubertal overweight children (age 9.0-14.6 years). RESULTS: After 4 years of therapy, height gain amounted to 1.8 SD; height (0.0 SD) and hand length (-0.2 SD) were normalised in the 2 prepubertal groups; in children above 6 years, height prediction approached parental target height. Weight for height rose in group 1 (to 0.64 SD) and decreased in group 2 (to 0.71 SD) to normal levels. Bone maturation of the pubertal children was too advanced to show a clear growth response to GH (height gain 0.42 SD). Even in this group, weight for height was reduced, but remained supernormal. CONCLUSION: Under exogenous GH, growth and body proportions are normalised in prepubertal children. With early institution of treatment, final height prediction reaches the parental target height range after 3 years. Such a growth-promoting effect of exogenous GH has so far only been described in children with GH deficiency. PMID- 11044803 TI - Energy expenditure, energy intake and prevalence of obesity after therapy for acute lymphoblastic leukemia during childhood. AB - OBJECTIVES: To investigate the prevalence and potential risk factors of obesity after therapy for childhood acute lymphoblastic leukemia (ALL). STUDY DESIGN: 39 ALL patients (age 10.7-20.5 years) who were in first remission for 3.4-14.6 years after standardized treatment with chemotherapy plus cranial irradiation (n = 25) or with chemotherapy alone (n = 14) were examined. After fasting overnight, the following parameters were investigated: body mass index (BMI) of patients and their parents; patients' BMI before ALL therapy; serum free thyroxin, growth hormone-dependent factors, estradiol, testosterone, cortisol, leptin and c peptide; fat-free mass (bioelectrical impedance); resting metabolic rate (RMR, indirect calorimetry); caloric intake (24-hour recall); and physical activity (questionnaire). RMR data were applied to the fat-free mass and compared with 83 controls. RESULTS: The prevalence of obesity (criterion: BMI > 2 SDS) was significantly (p < 0.05) higher after ALL therapy (38%; irradiated patients 48%, non-irradiated patients 21%) than before therapy (3%). Compared to non-irradiated patients, irradiated patients had significantly lower RMRs (-1.07 +/- 0.24 vs. 0.32 +/- 0.21 SDS; p < 0.05), reduced physical activity levels (1.41 +/- 0.03 vs. 1.52 +/- 0.03; p < 0.05), and lower concentrations of insulin-like growth factor binding protein-3 (-0.65 +/- 0.17 vs. 0.25 +/- 0.33 SDS; p < 0.05) and of free thyroxin (1.17 +/- 0.06 vs. 1.38 +/- 0.08 ng/dl; p < 0.05). Caloric intake was adequate. CONCLUSIONS: After ALL during childhood, patients face a higher risk of obesity. In the cranially irradiated patients, the likely causes are low physical activity, RMRs and hormonal insufficiency. PMID- 11044804 TI - Body composition abnormalities in children with Prader-Willi syndrome and long term effects of growth hormone therapy. AB - Obesity and hypothalamic GH deficiency contribute in different ways to the disturbances of body composition in Prader-Willi syndrome (PWS); while both increase the fat compartment, the reduction of lean tissue mass has been attributed mainly to GH deficiency. Therefore, body composition measured by dual energy X-ray absorptiometry was prospectively studied in 12 overweight children with PWS and weight for height (WfH) SDS >0 before and during 3.5 years of treatment with hGH (0.037 mg/kg/day) on average. In the long term, there is a net reduction of body fat from 3.1 to 1.2 SD, with a minimum at the end of the second year of treatment. WfH SDS correctly reflects body fat mass and its changes. The initial deficit of lean mass (-1.6 SD) is counteracted by GH only during the first year of therapy (increase to -1.25 SD). But in the long term, GH therapy does not further compensate for this deficit, when lean mass is corrected for its growth-related increase. In conclusion, exogenous GH changes the phenotype of children with PWS: fat mass becomes normal, but, at least in the setting studied, GH is not sufficient to normalize lean tissue mass. PMID- 11044805 TI - A novel nonsense mutation of the KAL gene in two brothers with Kallmann syndrome. AB - Kallmann syndrome (KS), defined by the association of hypogonadotropic hypogonadism and anosmia or hyposmia, can be caused by mutations in the KAL gene on Xp 22.3. This gene encodes an extracellular matrix glycoprotein called anosmin 1, which belongs to the class of cell adhesion molecules. In the absence of a functional KAL protein, migration of both olfactory and gonadotropin-releasing hormone neurons is arrested. A defective anosmin-1 molecule may also play a role in the development of synkinesia and renal agenesis, which are exclusively seen in the X-linked form of KS. We describe the clinical presentation and molecular diagnosis of the defect in two brothers with KS. An X-linked mode of transmission was assumed on the basis of synkinesia and the presence of oligomenorrhoea in the mother. A novel nonsense mutation was found in exon 13 of the KAL gene, encoding the region of the fourth fibronectin type III repeat of anosmin-1, which results in an apparently nonfunctional truncated protein. PMID- 11044806 TI - Enhancement of herpes simplex virus-induced polykaryocyte formation by 12-O tetradecanoyl phorbol 13-acetate: association with the reorganization of actin filaments and cell motility. AB - A morphological change induced by syn- herpes simplex virus type 1 (HSV-1), polykaryocyte formation, was enhanced by treatment with 12-O-tetradecanoyl phorbol 13-acetate (TPA) in A431 cells. TPA treatment decreased the number of stress fibers, but led to the development of spike-like filopodia and actin containing long projections. Similar reorganization of actin filaments was observed in HSV-1-induced polykaryocytes. The actin filament-disrupting drug cytochalasin D, but not the microtubule-disrupting drug nocodazole, inhibited the effect of TPA on polykaryocyte formation, indicating that the actin microfilament system plays a key role in this event. HSV-1 glycoprotein D (gD) was present in the cytoplasm of HSV-1-infected cells and gD gene-transfected cells; its expression became prominent at long cell projections in the presence of TPA. These findings suggest that the reorganization of actin filaments and cell motility are associated with the enhancing effect of TPA on HSV-1-induced polykaryocyte formation. PMID- 11044807 TI - Persistent infection mechanism of GB virus C/hepatitis G virus differs from that of hepatitis C virus. AB - OBJECTIVE: Changes in the deduced amino acid sequence of the envelope 2 (E2) region of the GB virus C/hepatitis G virus (GBV-C/HGV) were analyzed to investigate whether or not the region contributes to persistent infection with the virus. METHODS: Eight patients with acute hepatitis C and 1 patient with acute hepatitis of unknown etiology were included in the study. GBV-C/HGV RNA was detected in 6 patients, including the patient with hepatitis of unknown origin. The nucleotide sequence of the E2 region of hepatitis C virus (HCV) and GBV-C/HGV was determined by direct sequencing of polymerase chain reaction products in 5 patients with HCV infection and in 6 patients with GBV-C/HGV infection twice during the period of early infection and several months or years later in each patient. RESULTS: The mean substitution rate of the deduced amino acid sequence in the E2 region was over 100 times lower (p < 0.001) in GBV-C/HGV (0.01 +/- 0.04/month/100 sites) than in HCV (2.4 +/- 1.7/month/100 sites). The amino acid sequence of the loop domain of GBV-C/HGV-E2 did not change in any of the 6 patients. On the other hand, the sequence of the hypervariable region of HCV-E2 changed remarkably (5.9 +/- 4.3/month/100 sites). No amino acid substitution in the loop domain was observed in 7 additional patients who showed persistent GBV C/HGV viremia for more than 2 years. CONCLUSION: These results indicate that changes in the amino acid sequence of the E2 region are not involved in the mechanism of persistent GBV-C/HGV infection. PMID- 11044808 TI - Immunoglobulin G and M hepatitis C virus core antibody (JCC.2) response in chimpanzees infected experimentally with hepatitis C virus. AB - OBJECTIVE: To evaluate changes in serum immunoglobulin M (IgM) and G (IgG) hepatitis C virus (HCV) core antibodies after HCV infection in acute hepatitis C. METHODS: Serum HCV RNA and IgM and IgG HCV core antibodies were investigated using sera sequentially sampled from three chimpanzees experimentally infected with HCV. Serum IgG HCV core antibody titer was measured using a JCC.2 enzyme linked immunosorbent assay (ELISA) kit (Chemo-Sera-Therapeutic Research Center, Kumamoto, Japan). IgM core antibody titer was measured using horseradish peroxidase-labeled monoclonal anti-human IgM as the secondary antibody for the JCC.2 ELISA kit. Serum HCV RNA was detected using the 5' noncoding region as the primer according to the reverse transcriptase (RT) nested polymerase chain reaction (PCR) and competitive RT-PCR method. RESULTS: IgM JCC.2 antibody was detected when alanine aminotransferase (ALT) peaked, showing the closest correlation with the changes in ALT. A period during which IgM JCC.2 antibody was positive but HCV RNA as determined by RT-nested PCR was negative was observed after the elevation of ALT level. CONCLUSION: These results indicate the usefulness of detection of serum IgM JCC.2 antibody in making a definitive diagnosis of acute hepatitis C and the follow-up observation of hepatitis C. PMID- 11044809 TI - The hepatitis B virus pregenome: prediction of RNA structure and implications for the emergence of deletions. AB - The terminally redundant pregenomic RNA of human hepatitis B virus (HBV) comprises some 3,330 nucleotides and is a replicative intermediate in the production of the circular DNA genome. Deletions are known to arise in the HBV genome during the course of chronic infection and are sometimes associated with interferon therapy. These deletions are limited to small parts of the genome such as the 357-nucleotide pre-S1 region. Long RNA molecules such as the HBV pregenome have considerable structural flexibility and will undergo secondary structure shifts between energetically favourable states in a continuous and semi-random fashion. Since prediction of structure elements that are highly conserved in different forms of one RNA molecule is now feasible by computer modelling, we have analysed the whole HBV pregenome by two different RNA structure prediction algorithms and by new methods that exploit these algorithms. Significantly, the ends of pregenomic RNA were predicted to undergo both short-range and long-range interactions, which has relevance to our knowledge of the virus replicative cycle. By incorporating phylogenetic information relating to the 6 recognised genotypes of HBV, it was possible to highlight short secondary structures that may be common to all HBV strains. For example, although the pre-S1 region was predicted to undergo local folding of a loosely defined nature, most observed pre S1 deletions mapped to all or part of an arm carrying a better-defined structure. The loss of such sequences may be mechanistically attributable to polymerase skipping during reverse transcription, and the possible advantages of such deletions are considered. PMID- 11044810 TI - Multiple conformational epitopes are recognized by natural and induced immunity to the E7 protein of human papilloma virus type 16 in man. AB - The reactivity of sera from patients with cervical cancer with the E7 protein of human papilloma virus type 16 (HPV16) was estimated using a novel non-radioactive immunoprecipitation assay and four established protein- and peptide-based immunoassays. Six of 14 sera from patients with cervical cancer and 1 of 10 sera from healthy laboratory staff showed repeated reactivity with E7 in at least one assay. Four of the 7 reactive sera were consistently reactive in more than one assay, but only one was reactive in all four assays. Following immunization with E7, 2 of 5 patients with cervical cancer had increased E7-specific reactivity, measurable in one or more assays. No single assay was particularly sensitive for E7 reactivity, or predictive of cervical cancer. Mapping of E7 reactivity to specific E7 peptides was unsuccessful, suggesting that natural or induced E7 reactivity in human serum is commonly directed to conformational epitopes of E7. These results suggest that each assay employed in this study measures a different aspect of E7 reactivity, and that various reactivities to E7 may manifest following HPV infection or immunization. This finding is of significance for monitoring of E7 immunotherapy and for serological screening for cervical cancer. PMID- 11044811 TI - Relationship between five-year histological outcome and serial changes in serum alanine aminotransferase in patients with biochemical and virological relapse after interferon treatment for chronic hepatitis C. AB - OBJECTIVE: The aim of this study was to assess the correlation between serial changes in serum alanine aminotransferase (ALT) levels and histological outcome 5 years after treatment of patients with chronic hepatitis C with interferon (IFN). METHODS: We retrospectively evaluated 61 consecutive patients who underwent two liver biopsies, just before and 5 years after a 6-month course of IFN therapy, and who showed a relapse after therapy. The extent of liver fibrosis was estimated using a scale with seven grades. RESULTS: At the end of 6-month IFN therapy, 40 (65.6%) patients had normal serum ALT concentrations. However, 13 of the 40 patients relapsed within 6 months after IFN therapy. The average ALT during 5 years in 40 patients was less than or equal to 75 IU/l, while in the other 21 patients it was more than 75 IU/l. Among the 40 patients with a mean ALT less than 75 IU/l, 13 (33%) patients showed histological improvement, 26 (65%) showed no changes and only 1 patient (2%) showed worsening of liver fibrosis. On the other hand, among the 21 patients with a mean ALT of more than 75 IU/l, only 1 (5%) patient showed improvement, 8 (38%) showed no changes and 12 patients (57%) showed worsening of liver fibrosis. CONCLUSION: There is a significant relationship between 5-year histological outcome and serial changes in serum ALT in patients with biochemical and virological relapse after IFN treatment for chronic hepatitis C. PMID- 11044812 TI - Transcriptional pattern of varicella-zoster virus glycoproteins E and I. AB - Glycoproteins I (gI) and E (gE) of the Varicella-zoster virus are encoded by the neighbouring open reading frames 67 and 68. From earlier work it is known that both genes are transcribed into several transcript species which differ in size. From gI, three transcripts of 1.65, 2.7 and 3.6 kb and from gE, two transcripts of 2.15 and 3.6 kb in size are known. We present further northern analysis and show that these various transcript species appear in different amounts at different times after infection. 12 h after infection, transcripts of 1.65 kb (gI) and 2.15 kb (gE) were clearly detectable, whereas the other transcripts appeared later on. RT-PCR experiments using a set of seven different primers provided clear evidence that gI and gE are transcribed both, mono- and bicistronically with predominance on the respective monocistronic transcript. PMID- 11044813 TI - A vestigial X open reading frame in duck hepatitis B virus. AB - Duck hepatitis B virus (DHBV) appears to lack a homologue of the X protein found in mammalian hepadnaviruses. By replacing stop codons in the corresponding region of the DHBV genome, a hypothetical protein which closely matches the hydrophilicity profile of X proteins can be predicted, despite limited sequence homology. We conclude that a full-length X protein was once a common feature of the hepadnaviruses, conserved in structure but not sequence. PMID- 11044814 TI - Segmental neurofibromatosis in association with nevus sebaceus of Jadassohn. AB - We describe an unusual case involving the simultaneous occurrence of segmental neurofibromatosis (Type V NF) in a patient with a large nevus sebaceus of Jadassohn in the same physical distribution. Causative mechanisms of development of these 2 genetic disorders have not been definitively linked. Factors producing these diseases probably involve similar tissues at the same point in development because both have been reported in association with central nervous system anomalies and have been classified among the neurocutaneous syndromes. This is a case of a nevus sebaceus occurring in association with and in the same physical distribution as segmental NF. These disorders most likely represent a spectrum of disease within the phakomatoses. PMID- 11044815 TI - Stevens-Johnson syndrome associated with concomitant use of lamotrigine and valproic acid. AB - Lamotrigine (LTG) is a novel antiepileptic drug effective in partial and generalized seizures. Recently, this drug has started being used for mood stabilization in psychiatric patients. Cutaneous side effects of this drug are mostly maculopapular eruptions that have been seen in 3% to 10% of patients. We describe a 33-year-old female patient in whom Stevens-Johnson syndrome (SJS) developed because of LTG. The drug was discontinued, and patient's signs and symptoms progressively resolved in 10 days after oral prednisolone therapy. The case is relevant because this is the first case of SJS as a result of LTG, probably associated with concomitant use of valproic acid and neuroleptic drugs. PMID- 11044816 TI - Mast cells in an angiosarcoma complicating xeroderma pigmentosum in a 13-year-old girl. AB - A cutaneous angiosarcoma, a rare tumor that occurs almost exclusively in sun exposed skin of individuals older than 50 years, developed in an adolescent with Xeroderma pigmentosum (XP). As this is the second report of a child with angiosarcoma and XP, ultraviolet-induced DNA damage may be involved in the pathogenesis of this tumor. Strongly increased numbers of mast cells were found, particularly in the peripheral tumor area, which may reflect with the requirement of mast cells for the growth of vascular structures or a role for mast cells in the antitumor immune response. PMID- 11044817 TI - A case of extragenital lichen sclerosus following Blaschko's lines. AB - Extragenital lichen sclerosus is most common on the neck, shoulders, and upper portion of the trunk. A 20-year-old Korean man had asymptomatic, well-demarcated, linear violaceous plaques on the left lateral chest, in a pattern corresponding to the lines of Blaschko. We describe a case with a pattern of extragenital lichen sclerosus following Blaschko's lines. PMID- 11044818 TI - Kimura's disease with oral ulcers: response to pentoxifylline. AB - We describe a man with Kimura's disease whose presentation included lymphadenopathy and cutaneous nodules, but was most distinctive for painful oral ulcerations. His lesions showed an initially moderate, but ultimately minimal response to monthly triamcinolone injections. With oral pentoxyifylline, he showed resolution of all of his lesions for 14 months. On cessation of his treatment, his disease flared for 3 months. When pentoxyifylline was restarted, his lesions regressed again within 4 weeks. We review the literature on Kimura's disease. PMID- 11044819 TI - An unusual presentation of dermatomyositis: the type Wong variant revisited. AB - We describe a 53-year-old white woman with dermatomyositis (DM) who had additional clinical findings of pityriasis rubra pilaris (type Wong dermatomyositis) with histopathologic features of both pityriasis rubra pilaris (PRP) and porokeratosis. Type Wong dermatomyositis was originally described in 11 patients by Wong in 1969 and has been reported in 5 additional patients. This is a rarely described phenomenon in which patients with DM develop cutaneous hyperkeratotic lesions that resemble PRP and histologically show follicular hyperkeratosis and hair follicle destruction. Arrector pilorum muscles also show degenerative findings and myositis. We believe that this is the first reported case of a patient with type Wong DM who also has clinical and histologic features suggestive of porokeratosis. This is important because of the association of adult-onset dermatomyositis with internal malignancy and the well-documented association of porokeratosis with immunosuppression. These clinical and histologic findings serve as markers for malignancy in patients with DM. These patients warrant a complete review of systems and investigation for age appropriate neoplasms as well as close long-term follow-up by dermatologists to ensure that these cutaneous eruptions are not overlooked. PMID- 11044820 TI - An annular plaque due to Mycobacterium haemophilum infection in a patient with AIDS. AB - Infections with mycobacteria are of importance in the differential diagnosis of skin lesions in immunocompromised patients. We report the case of a human immunodeficency virus (HIV)-infected patient who presented with an annular plaque as cutaneous manifestation of Mycobacterium haemophilum. Improvement of the immunologic status is important for successful treatment. Complete resolution was only observed when the antimycobacterial therapy was combined with antiretroviral therapy. PMID- 11044821 TI - Papular-purpuric gloves and socks syndrome in HIV-positive patients. AB - Three HIV-positive women showed clinical signs of papular-purpuric gloves and socks syndrome and serologic evidence of acute Parvovirus B19 infection. The course of the disease was complicated by anemia and persistent skin lesions, probably related to inadequate immune response. Because anemia in AIDS patients may be due to many causes, the history of recent Parvovirus B19 infection is helpful in suggesting the etiologic diagnosis. PMID- 11044822 TI - Chronic hepatitis C virus infection associated with a generalized granuloma annulare. AB - We report the first case of generalized granuloma annulare occurring in a 65-year old woman who was chronically infected with hepatitis C virus. Granuloma annulare totally regressed during alpha-interferon treatment. As chronic hepatitis C virus infection is frequent, a serodiagnosis would be of interest in patients who have generalized granuloma annulare. A link between these 2 diseases may be strongly suspected. PMID- 11044823 TI - Delayed-type skin reaction to the heparin-alternative danaparoid. AB - Eczematous, infiltrated plaques at the site of subcutaneously administered heparin appear to be common. Heparinoids cannot be recommended in general as a substitute for heparin or low molecular weight heparin because delayed-type skin reactions to these molecules can also occur, as demonstrated in this case report. PMID- 11044824 TI - Rapid response of IgA pemphigus of subcorneal pustular dermatosis type to treatment with isotretinoin. AB - Diagnosing IgA pemphigus and distinguishing between its 2 subtypes, intraepidermal neutrophilic IgA dermatosis type and subcorneal pustular dermatosis type, is important because treatment of IgA pemphigus has to be different from treatment of other blistering autoimmune dermatoses. We present a patient with subcorneal pustular dermatosis type of IgA pemphigus who rapidly responded to systemic treatment with isotretinoin. Specific diagnosis was established by detecting IgA serum activity to desmocollin 1 by indirect immunofluorescence microscopy on unfixed COS7 cells transfected with desmocollin 1. No IgA or IgG serum reactivity was found to recombinant forms of desmogleins 1 and 3 by an antigen-specific enzyme-linked immunosorbent assay. The disease was not effectively controlled by conventional therapeutic regimens. Systemic treatment with isotretinoin 20 mg daily led to complete clearance of skin lesions within 3 weeks. Assaying IgA serum reactivity to desmocollin 1, desmoglein 1, and desmoglein 3 as a valuable method for establishing the diagnosis and differentiating the 2 subtypes of IgA pemphigus. Isotretinoin was an effective drug in the treatment of subcorneal pustular dermatosis type of IgA pemphigus in this patient. PMID- 11044825 TI - Two cases of nevus comedonicus: successful treatment of keratin plugs with a pore strip. AB - Nevus comedonicus is an uncommon condition of unknown cause. We used a pore strip cosmetic pack to eliminate the keratin plugs in 2 patients with nevus comedonicus, producing excellent results. PMID- 11044826 TI - Aortic angiosarcoma presenting with cutaneous metastasis: case report and review of the literature. AB - Aortic angiosarcoma is a rare malignancy. Clinical diagnosis is difficult, as the presenting symptoms mimic more common aortic lesions, particularly atherosclerosis. Dermatologists and dermatopathologists play a critical role in recognizing cutaneous metastases as a manifestation of this life-threatening tumor. We describe the fourth case of aortic angiosarcoma in the literature with initial presentation in the skin. PMID- 11044827 TI - Acquired dermal melanocytosis in an African-American: A case report. AB - We report an unusual case of acquired dermal melanocytosis occurring in a 57-year old African-American woman. Macular pigmentation over the patient's cheeks, back, and sclerae developed over the course of 18 months without identifiable antecedent. The patient was taking only hormone replacement therapy and was generally healthy. Histologic examination of the skin revealed dendritic melanocytes and melanophages in the mid- to upper dermis. We briefly review the literature surrounding acquired dermal melanocytosis and continue discussion regarding its pathogenesis. PMID- 11044828 TI - Capillaritis associated with interferon-alfa treatment of chronic hepatitis C infection. AB - We report the case of a 36-year-old man who was referred with an asymptomatic eruption that started on both lower legs. This started shortly after being commenced on interferon-alfa for chronic hepatitis secondary to hepatitis C. Clinically, the eruption was consistent with a capillaritis (pigmented purpuric dermatosis). Histology confirmed this to be lymphocytic vasculitis. Lymphocytic vasculitis is frequently identified in the salivary glands of patients who are hepatitis C positive. Although leukocytoclastic vasculitis confined to the skin is frequently reported with hepatitis C, lymphocytic vasculitis is rarely reported. We consider that the lymphocytic vasculitis in our patient occurred as a result of interferon-alfa treatment because of the strong temporal relationship between the onset of the skin eruption and drug therapy. PMID- 11044829 TI - Ectopic respiratory mucosa in the skin associated with skeletal malformation and polydactyly. AB - We report a 19-year-old man with congenital erythematous plaque on his left arm that was found histopathologically to be composed of respiratory mucosa. The patient had a triphalangeal thumb and polydactyly in the left hand. This is, to our knowledge, only the third case of ectopic respiratory epithelium presenting as a superficial lesion in the skin to be reported in the English literature, and the first case associated with skeletal malformation and polydactyly. PMID- 11044830 TI - Localized granuloma annulare and autoimmune thyroiditis: a new case report. AB - We present further evidence that granuloma annulare and autoimmune thyroiditis may be associated. Only 11 cases have been previously reported with this association as far as we know. It may be advantageous to consider checking the thyroid-stimulating hormone and/or antithyroid antibodies before administering medications that could affect thyroid function in patients with granuloma annulare. PMID- 11044831 TI - Nonerythrodermic, leukemic variant of cutaneous T-cell lymphoma with indolent clinical course: Th2-type tumor cells lacking T-cell receptor/CD3 expression and coinfiltrating tumoricidal CD8 T cells. AB - As typically represented by Sezary syndrome, the leukemic form of cutaneous T cell lymphoma (CTCL) mostly exhibits erythroderma. A patient with CTCL had slowly developing skin tumors as well as chronic leukemia. The tumor cell was CD4+ CD7- Th2 cells lacking T-cell receptor/CD3 complex and persistently occupied 27% to 48% of peripheral blood lymphocytes. In skin tumors, only 13% of tumor infiltrating lymphocytes were malignant cells and substantial numbers of nonmalignant CD4+ or CD8+ T cells and B cells coinfiltrated. CD8+-infiltrating T cells had cytotoxic activity against the malignant T cell. Our case demonstrates the existence of the leukemic form of CTCL presenting with skin manifestation other than erythroderma and parapsoriatic patches. The nonerythrodermic feature and indolent course may be associated with the lack of T-cell receptor/CD3 expression and coinfiltration of a high percentage of nontumor lymphocytes, including tumoricidal CD8+ T cells. PMID- 11044832 TI - Annular leukocytoclastic vasculitis associated with monoclonal gammopathy of unknown significance. AB - Leukocytoclastic vasculitis is a condition characterized by necrotizing neutrophilic inflammation of small dermal blood vessels usually resulting in palpable purpuric lesions. Leukocytoclastic vasculitis may be secondary to a variety of medications and underlying disease processes, including infections, connective tissue disorders, and malignancies. We describe a patient with a monoclonal gammopathy of unknown significance in whom leukocytoclastic vasculitis developed, manifested by a few prominent annular plaques on the lower extremity. The rare association between monoclonal gammopathy of unknown significance and leukocytoclastic vasculitis as well as the unusual annular presentation of the disease in this patient is discussed, and the relevant literature is reviewed. PMID- 11044833 TI - Localized pruritus: a presenting symptom of a spinal cord tumor in a child with features of neurofibromatosis. AB - Central nervous system (CNS) and spinal cord tumors are not uncommon in patients with neurofibromatosis (NF); however, it is impossible to select patients with NF who are at a particularly high risk. Localized pruritus may be a clue to the presence of a spinal cord or CNS tumor. This is the first report of an infant with features of NF, whose presenting symptom of a spinal cord tumor was localized symmetrical dermatomal itch. Moreover, we review the literature of localized pruritus in CNS and spinal cord tumors and peripheral nervous system conditions. PMID- 11044834 TI - Primary cutaneous plasmacytoma: report of a case with review of the literature. AB - Primary cutaneous plasmacytoma (PCP) is a rare cutaneous B-cell lymphoma. We report a new case of PCP and review data published in the literature. A 55-year old man presented with 2 erythematous plaques on the upper trunk, showing histologic and immunohistochemical features of PCP. Staging investigations excluded extracutaneous manifestations of the disease. The patient was treated with melphalan and prednisone associated with local radiotherapy. Twenty-four months after the first presentation he is alive and well. Only 29 cases of PCP have been reported in the last 50 years. The main prognostic factor seems to be the clinical presentation (solitary vs multiple lesions). Solitary lesions of PCP may be treated conservatively by surgical excision or local radiotherapy. PMID- 11044835 TI - Synchronous development of disseminated superficial porokeratosis and hepatitis C virus-related hepatocellular carcinoma. AB - Immunosuppression and transplantation have been reported to induce porokeratosis (PK), especially its variant, disseminated superficial PK (DSP). On the other hand, there is ample evidence of a relationship between hepatitis C virus (HCV) infection, liver cirrhosis (LC), and hepatocellular carcinoma (HCC). We report 3 cases of DSP in which the outbreak of DSP was suspected to have occurred during the development of HCC in patients with HCV-positive LC. The patients had undergone ultrasonographic study regularly, and no signs of malignancy had been found before the development of DSP. Their outbreaks of DSP were very acute, and the period between the development of DSP and diagnosis of HCC ranged from 2 to 6 months. The association of HCV-related HCC and DSP has never been previously reported. HCV-induced immunomodulation or its effect on the p53 system may be the basis for this type of association. It is necessary to consider development of HCC whenever DSP is found in HCV-positive patients. DSP may be a new paraneoplastic dermadrome. PMID- 11044836 TI - Early onset sarcoidosis masquerading as juvenile rheumatoid arthritis. AB - Symptoms of early onset sarcoidosis characterized by skin eruptions, arthritis, and uveitis mimic those of systemic type juvenile rheumatoid arthritis (JRA). We report 2 Japanese patients with early onset sarcoidosis, both of whom were initially diagnosed and treated as having JRA. Intermittent fever and synovial swelling may mask sarcoidosis in children less than 4 years old. PMID- 11044837 TI - Telangiectasia macularis eruptiva perstans and multiple myeloma. AB - The association of mast cell diseases and some hematologic malignancies, usually myeloproliferative disorders, myelodysplastic syndromes, and acute leukemia is well recognized. We report the case of a patient with telangiectasia macularis eruptiva perstans, a rare form of cutaneous mastocytosis, and multiple myeloma, an association that has been described only twice in the literature. Parallel improvement of both conditions was observed under chemotherapy regimens for multiple myeloma. Pathogenesis remains unclear, although the abnormalities in the c-kit pathway may play a role in the proliferation of cells from both lineages. PMID- 11044839 TI - Brazilian society of oral rehabilitation - founded in 1977 PMID- 11044840 TI - The japan prosthodontic society - founded in 1933 PMID- 11044838 TI - US newborn screening system guidelines II: follow-up of children, diagnosis, management, and evaluation. Statement of the Council of Regional Networks for Genetic Services (CORN). PMID- 11044841 TI - The australian prosthodontic society inc-founded in 1961 PMID- 11044842 TI - Fabrication of a facial shield to prevent facial injuries during sporting events: a clinical report. PMID- 11044843 TI - Neutral zone approach for denture fabrication for a partial glossectomy patient: a clinical report. AB - According to the neutral zone concept, an appropriate denture form can be molded into a physiologic tooth arrangement by using a narrow rim bar occlusion, tissue conditioner, and a direct relining technique for both intaglio and cameo surfaces by using VLC reline resin. Denture stability can be obtained with this "border molding" technique, not only for edentulous patients but also for those with oral deformities. PMID- 11044844 TI - Microleakage study of various soft denture liners by autoradiography: effect of accelerated aging. AB - STATEMENT OF PROBLEM: The longevity of soft denture liners is a major clinical problem. Debonding of the soft liner from the denture base material is one of the factors that influence their longevity. Debonding of the soft liner can be attributed to microleakage at the interface. PURPOSE: This study investigated microleakage at the interface of various soft liners and base materials. MATERIALS AND METHOD: Six soft liners were investigated. Forty specimens of each material in disk form (10 mm in diameter, thickness of approximately 4 mm) were prepared. Twenty specimens of each material were stored in an accelerated weathering tester for 900 hours. For 2 days, all disks were immersed in (45)Ca radioisotope solution, then they were embedded in acrylic resin blocks and sectioned longitudinally. Autoradiographic imaging was used to determine microleakage at the interface of the soft liners and their bases. RESULTS: Significant differences between nonaged materials were found (P<.05).The difference between Molloplast B and Mucopren (silanized) was not significant (P<.05). Differences among aged materials were significant (P>.05). Differences between Mucopren (nonsilanized), Mucopren (silanized), and Ufigel P-Tokuyama were not significant (P<.05). Significantly decreased microleakage characteristics were determined for Molloplast B, Mucopren (nonsilanized) and Ufigel P liners after aging. CONCLUSION: Microleakage of Mucopren and Molloplast B lining materials was the lowest. However, the microleakage of Flexor and Simpa was the highest. The aging process did not significantly affect the microleakage characteristics of the Simpa, Flexor, Mucopren (silanized), or Tokuyama materials. Molloplast B, Mucopren (nonsilanized), and Ufigel P materials should significantly decrease microleakage properties after aging. PMID- 11044845 TI - All-ceramic surveyed crowns for removable partial denture abutments. AB - All-ceramic restorations may have limitations when used in combinations with other treatment modalities. Removable partial denture (RPD) abutment crowns are typically shaped to provide guide planes, rest seats, and retentive areas. Porcelain-to-metal crowns are routinely fabricated with retentive contours in the veneering porcelain, but until recently, the contours of the other portions of the restoration were developed in metal. This article describes the fabrication of all-ceramic crowns for RPDs with rest seats and guide planes in densely sintered aluminum oxide and retentive areas in veneering porcelain. Within the limitations outlined, this procedure allows the practitioner to use all-ceramic crowns in situations previously reserved for metal or metal-ceramic restorations. PMID- 11044846 TI - Accuracy of casts generated from dual-arch impressions. AB - STATEMENT OF PROBLEM: Dual-arch trays are often used to make simultaneous impressions of a prepared tooth and the opposing teeth. Many dentists are concerned with the accuracy of the casts generated from this type of impression. PURPOSE: The purpose of this investigation was to compare the accuracy of stone casts of a prepared tooth generated using 2 types of dual-arch impression tray/impression material combinations. MATERIAL AND METHODS: The maxillary left first molar on a dentoform mounted on an articulator was prepared for a full coverage gold crown. Ten impressions were made with either a plastic (P) or metal (M) tray and a polyether (PE) or vinyl polysiloxane (VPS) impression material. Each impression was cast in improved dental stone, and the buccolingual dimension of the die was measured at the midpoint of the buccal and lingual gingival margins. The prepared tooth (T) served as the control. The data were analyzed using a 2-factor analysis of variance (ANOVA) alpha=.05. RESULTS: The P/VPS combination (10.673 mm) produced the largest die, followed by P/PE (10.602), T (10.508), M/PE (10.484), and M/VPS (10.472). The 2-factor ANOVA showed a significant difference between the tray types but not between the impression materials. CONCLUSION: The metal trays produced dies smaller than the tooth, and the plastic trays produced dies that were larger. PMID- 11044847 TI - Shear bond strength of "one bottle" dentin adhesives. AB - STATEMENT OF PROBLEM: Several 1-bottle dentin adhesive resins have been introduced to the market; however, their shear bond strength still requires further investigation. PURPOSE: This study was designed to evaluate the shear bond strength of five 1-bottle fifth generation dentin adhesive resin materials. MATERIAL AND METHODS: Bond 1, Single Bond, One-Step, Prime & Bond 2.1, and Tenure Quick with fluoride were evaluated. Tenure All-Surface Bonding System, a fourth generation dentin adhesive resin, was used as the control group. Twelve specimens were prepared from each material, and the shear bond strength was measured by using a universal testing machine at a crosshead speed of 0.5 mm/min. Fracture patterns were studied with the use of light and SEM. The results were analyzed with the use of ANOVA. RESULTS: The mean shear bond strength for each resin was as follows: Bond 1: 18.36 +/- 3.19 MPa; Single Bond: 16.22 +/- 2.11 MPa; One Step: 22.51 +/- 3.69 MPa; Prime & Bond: 16. 64 +/- 3.66 MPa; Tenure Quick: 16.43 +/- 3.2 MPa; and Tenure All-Surface Bonding System: 15.06 +/- 3.5 MPa. The shear bond strength values of One-Step dentin adhesive resin showed significant differences compared with the other 5 groups (P<.001). No significant differences were seen between the control group and the other four 1-bottle dentin adhesive resins (0.25 > P>.1). Eighty percent or more of the specimens for each adhesive failed at the dentin/adhesive interface. An exception was Tenure All-Surface Bonding System in which all specimens failed at the dentin/adhesive interface. CONCLUSION: The 1-bottle systems tested bond to dentin with a strength similar to that of the control group. In addition, the shear bond strengths of 4 of the 1 bottle systems tested were not significantly different. PMID- 11044848 TI - Resin-bonded, glass fiber-reinforced composite fixed partial dentures: a clinical study. AB - STATEMENT OF PROBLEM: Resin-bonded, glass fiber-reinforced composite fixed partial dentures (FPDs) have been under development for some time. There is a lack of data regarding the clinical usefulness of such prostheses. PURPOSE: The clinical performance of 31 resin-bonded, glass fiber-reinforced composite fixed partial dentures was evaluated in a preliminary study. MATERIAL AND METHODS: The prostheses were made to replace 1 to 3 missing maxillary or mandibular teeth in each of 31 patients. The prostheses had a framework made of continuous unidirectional E-glass fibers with multiphase polymer matrix and light polymerized particulate composite resin veneering. The prostheses were examined after 6-month periods for up to 24 months (mean follow-up time was 14 months). Partial or total debonding of the prostheses or the framework fracture was considered a treatment failure. RESULTS: Two prostheses debonded during the follow-up period; 1 debonding was related to improper occlusal adjustment and the other to unknown reasons. Kaplan-Meier survival probability at 24 months was 93%. No framework fractures were observed. CONCLUSION: The results of this preliminary study suggest that the resin-bonded, glass fiber-reinforced FPDs may be an alternative for resin-bonded FPDs with a cast metal framework. PMID- 11044849 TI - Clinical fit of Procera AllCeram crowns. AB - STATEMENT OF PROBLEM: Color stability, strength, and accuracy of fit are the main requirements for complete-ceramic crowns. The Procera AllCeram crown system is a CAD/CAM system used to fabricate individual complete-ceramic crowns that have a dry sintered, aluminum oxide core and appear to match clinical requirements. However, there are few articles about the clinical fit of all-ceramic crowns. PURPOSE: This in vivo study measured the accuracy of fit of Procera AllCeram crowns in anterior and posterior teeth. MATERIAL AND METHODS: The clinical fit of 80 anterior and posterior Procera AllCeram crowns was evaluated by a replica technique with a light body silicone to fill space between crown and tooth and a heavy body silicone to stabilize the light body film. After removal from the artificial crowns, the replicas were segmented, and measurements of the film thickness were performed with a light microscope. RESULTS: Medians of mean marginal gap widths were between 80 and 95 microm in anterior teeth and between 90 and 145 microm in posterior teeth. Medians of maximal marginal gap widths ranged from 80 to 180 microm in anterior teeth and from 115 to 245 microm in posterior teeth. CONCLUSION: The accuracy of fit achieved by Procera AllCeram was comparable to other conventional and innovative systems. PMID- 11044850 TI - Double-layer porcelain veneers: effect of layering on resulting veneer color. AB - STATEMENT OF PROBLEM: Densely sintered aluminum oxide veneered with feldspathic porcelain seems to be a promising technology for the fabrication of porcelain laminate veneers, which provide both strength and esthetics. To effectively use this approach for porcelain veneers, practitioners should know how the resulting color is affected by adding a layer of veneering porcelain onto the aluminum oxide. PURPOSE: This study compared changes in CIE L*a*b* color coordinates of simulated stained teeth when covered with the aluminum oxide core disks alone and after the disks had been veneered with 3 different shades of porcelain. MATERIAL AND METHODS: Fifteen aluminum oxide disks were divided into 3 groups. Each of 5 disks was veneered with porcelain of the Vita shades A1, A2, and B4, respectively. The colors of the substrate covered with the nonveneered disks and the veneered disks were measured separately, and the color differences were calculated. Measurements of the disks on a white background were also performed and used as controls. The extracted data were compared with Vita shade values available from the literature. RESULTS: Statistically significant differences in color coordinates of dark substrates were recorded between the substrate covered by an aluminum oxide disk and the addition of veneering porcelain to the aluminum oxide disks. Veneering the aluminum oxide disks with A1, A2, and B4 porcelain produced significant differences in the resulting color when placed on a dark substrate. The direction of the color modifications correlated with values obtained from the literature. The resulting color was also affected by the color of the underlying structures. CONCLUSION: Although aluminum oxide alone does have a degree of masking capability, the resulting color of porcelain veneers with the use of this material can successfully be modified with the veneering porcelain. PMID- 11044851 TI - Implant-retained cantilever fixed prosthesis: where and when. AB - STATEMENT OF PROBLEM: Many clinical situations are suitably treated with cantilevered implant-supported cemented fixed restorations. PURPOSE: This article details the indications for the restorative dentist to use a cantilever fixed prosthesis after insertion of ITI dental implants. CONCLUSION: The use of implants to support cantilevered fixed partial dentures has been successful in selected clinical situations. PMID- 11044852 TI - Correlation of noncarious cervical lesion size and occlusal wear in a single adult over a 14-year time span. AB - STATEMENT OF PROBLEM: Noncarious cervical lesions are described as having a multifactorial cause, with occlusal trauma and toothbrush abrasion frequently mentioned as major factors. Finite element modeling studies have demonstrated a relocalization of occlusal stresses to the cervical area due to flexure of the crown. This may cause microcracking, especially under tensile stresses, that will lead to a loss of enamel and dentin in the cervical region. Clinical confirmation of an occlusal cause for noncarious cervical lesions has been difficult to obtain. PURPOSE: This study investigated whether occlusal wear was correlated with an increase in the size of noncarious cervical lesions. MATERIAL AND METHODS: Loss of contour at occlusal and cervical sites on 3 teeth of a single individual was measured using digital and visualization techniques at 3 time intervals over a 14-year time span. The 1983 baseline casts and 1991, 1994, and 1997 clinical impressions of a single adult patient with existing noncarious cervical lesions were replicated in epoxy. Surfaces of all replicas were digitized with a contact digitizing system. Sequential digitized surfaces were fit together and analyzed using AnSur-NT surface analysis software. Clinical losses of surface contour by volume and depth of the left mandibular first molar and first and second premolars were recorded. RESULTS: Nine measurements of cervical volume loss (range 0.9 to 11.5 mm(3)) and 9 corresponding measurements of occlusal volume loss (range 0.39 to 7.79 mm(3)) were made. The correlation between occlusal and cervical volume loss was strong (r(2)=0.98) and significant (P<.0001). CONCLUSION: For the single adult patient in this study, there was a direct correlation between occlusal wear and the growth of noncarious cervical lesions. PMID- 11044853 TI - Changes in the mechanical properties and surface texture of compomer immersed in various media. AB - STATEMENT OF PROBLEM: Limited information is available about the mechanical behavior of compomer under intraoral conditions. PURPOSE: This in vitro study evaluated changes in the mechanical properties and surface texture of compomer and other materials, used in similar clinical circumstances, when immersed in various media. MATERIAL AND METHODS: Hardness measurement for 5 tested materials and 4 immersion media was obtained with a Vickers hardness testing machine. Compressive strength was measured using an Autograph at a crosshead speed of 0.75 mm/min. All readings were taken for up to a 60-day period. An electron probe microanalyzer was used to give an SEM image. RESULTS: The average compressive strength and Vickers surface hardness showed a significant difference between materials. Results showed an overall increase in the solubility of specimens immersed in low pH soft drinks. CONCLUSION: There was a difference in the mechanical properties and surface texture of the materials tested in this study when they were immersed in various media. PMID- 11044854 TI - Irritation test of tissue adhesives for facial prostheses. AB - STATEMENT OF PROBLEM: Adhesives are commonly used to improve the retention of a facial prosthesis to the skin. Although no requirement exists for facial prosthetic adhesives, an adhesive should be nonirritating and nontoxic. PURPOSE: This study assessed the irritative potential of facial prosthetic adhesives by using an in vitro technique for detection of eye-irritating chemicals. MATERIAL AND METHODS: Ten adhesives were evaluated by the hens egg test chorioallantoic membrane method. Adhesives were applied to the chorioallantoic membrane in fertilized hen eggs, and the membrane examined by a photomacroscope for injury to the blood vessels. The average irritation score was calculated from the recorded times for the debut of hemorrhage, lysis, and coagulation, and the products were classified as being non, slight, moderate, or strong irritants, based on the irritation score. RESULTS: The predominant injury to the membrane was coagulation of blood vessels, and the exposure time needed to initiate the reaction was dependent on the composition of the product. Four products were classified as strong irritants, 1 as moderate, and the remaining 5 as slight or nonirritant. CONCLUSION: On the basis of a test for eye irritation, the irritant potential of tissue adhesives varied from non to severe. The most severe reactions were mainly seen in products containing the solvent ethyl acetate. PMID- 11044855 TI - Functional impression technique in the management of an unusual facial defect: a clinical report. PMID- 11044856 TI - Analysis of lateral tooth movement during forced orthodontic eruption. AB - STATEMENT OF PROBLEM: Forced eruption is a prosthodontic procedure that enables the treatment of otherwise problematic restorative conditions. During the vertical orthodontic movement, the root may be moved laterally, affecting the position of a tooth in the arch. PURPOSE: This study quantified the degree of lateral movement possible during the eruptive procedure and addressed the significance of this movement from a theoretical and clinical standpoint. MATERIAL AND METHODS: A patient treatment, which demonstrated the movement in a single direction on the buccolingual axis, was isolated. On the basis of this theoretical model, a clinically relevant model was developed: A "worst-case" situation for each posterior tooth was calculated, from which clinical conclusions may be derived. RESULTS: . In a given extrusion angle of 30 degrees, a lateral movement/shift of 0.58 mm resulted per 1 mm of eruption distance. The lateral movement for every millimeter of eruption and the maximum extrusion angle for each of the maxillary and mandibular posterior teeth were calculated. CONCLUSION: The lateral movement that accompanies the forced eruption procedure may compromise or be used to esthetically enhance prosthodontic treatment. PMID- 11044857 TI - Diagnostic management of interdental spacing. AB - It is a challenge to restore function and esthetics for patients with excessive interdental spacing. This article describes an alternative diagnostic procedure to permit clear communication of the intended outcome among the restorative dentist, orthodontist, and patient. This procedure uses fabrication of multiple duplicated casts, sectioned stone teeth, repositioned stone teeth, full-contour diagnostic wax-ups, and occlusal putty impressions. This procedure can be used as proposed or for various situations in which complex restorative treatment is needed. PMID- 11044858 TI - Laboratory considerations in rotational path removable partial dentures. AB - Clinical indications and contraindications have been well covered in the literature for rotational path removable partial dentures (RPDs). However, only minimal coverage has been devoted to problems encountered that may prevent the proper fabrication of these restorations by dental technicians. The purpose of this article is to illustrate 2 problems that dental technicians occasionally encounter that make the fabrication of rotational path RPDs difficult or impossible. Design modifications by the clinician can eliminate 2 problems faced by technicians fabricating rotational path RPDs. PMID- 11044859 TI - Smoothing of soft lining materials with chloroform-based varnish. PMID- 11044860 TI - Conditioned changes in ultrasonic vocalizations to an aversive olfactory stimulus are lateralized in 6-day-old rats. AB - Using a soft rubber plug to block airflow in one naris, Kucharski, Johanson, and Hall (1986) found that some forms of olfactory memory (e.g., odor preferences) were lateralized in young rats while other forms (e.g., conditioned activation and mouthing) were not. The present experiments extended that research by showing that conditioned increases in ultrasonic vocalizations were also lateralized. That is, when exposed to an odor that was previously paired with footshock, 6-day old rats significantly increased their rate of vocalizing. This response only occurred, however, when the naris open at training was also open at test. The use of the developing rat as a natural split-brain preparation appears to be an effective procedure with which to broaden current approaches to the analysis of learning, memory, and emotion. PMID- 11044862 TI - Olfactory conditioning facilitates diet transition in human infants. AB - We evaluated whether Pavlovian conditioning methods could be used to increase the ingestion of non-preferred solutions by formula-fed human infants. In baseline measures, 5-7 month old infants sucked less frequently and consumed less water than regular formula. During a 3-day olfactory conditioning period, parents placed a small scented disk, the conditioned stimulus, on the rim of their infants' formula bottle at every feeding. Following this training, infants' responses to water were tested when their water bottles had a disk scented with the training odor, a novel odor, or no odor. Infants tested with the training odor sucked more frequently and consumed significantly more water than they had at baseline. Infants tested with no odor or a novel odor consumed water at or below baseline levels. These data demonstrate that olfactory conditioning can be used to enhance ingestion in infants and suggest that such methods may be useful for infants experiencing difficulty when making transitions from one diet to another. PMID- 11044861 TI - The first suckling episode in the rat: the role of endogenous activity at mu and kappa opioid receptors. AB - The present study examined the role of endogenous activity at mu and kappa opioid receptors in attachment to and ingestion of milk from a surrogate nipple in cesarean-delivered newborn rats prior to regular suckling experience. Selective opioid antagonist drugs were injected into the cisterna magna (IC administration) or lateral ventricles (ICV administration). Blockade of endogenous activity at mu opioid receptors by IC administration of the selective antagonist CTOP reduced attachment time and markedly increased disengagements from the nipple. CTOP also increased the intensity of suckling measured as milk intake per min attached to the nipple, when milk was available from the nipple in a free-access regime, and enhanced intake when milk was infused through an intraoral cannula aside from the suckling context. The ICV administration of the selective kappa antagonist nor BNI considerably increased latency to grasp the surrogate nipple, while time on the nipple and milk intake were decreased. The presented data suggest that populations of mu and kappa receptor-containing neurons, differentiable by the route of antagonist administration, play an important role in initiation and maintenance of suckling behavior in the newborn rat during its first encounter with the nipple and milk. The kappa opioid system is predominantly involved in the initiation of the newborn's behavior directed toward the nipple providing milk. The role of the mu opioid system seems more complicated: it transforms initial oral grasp responses into sustained attachment to the nipple and maintains the intake of milk at a certain physiological level. PMID- 11044863 TI - Dissociations between psychobiologic reactivity and emotional expression in children. AB - Although there are general assumptions that physiological and behavioral indices of emotion are interrelated, empirical research has revealed inconsistent findings with regard to their degree of association, particularly in children. Two studies were conducted to examine the relations between cardiovascular reactivity and emotional behavior. In the first study, 3- to 6-year-olds completed challenging tasks during which measures of their physiological responses and facial expressions were obtained. With age, children's heart rate decreased, vagal tone increased, and facial expressions became slightly more exaggerated. However, children's physiologic reactions were unrelated to their concurrent facial expression when all children were considered, when only boys were considered, and when children extreme in their physiologic reactions were considered. Only among girls was physiologic reactivity moderately associated with concurrent negative expressiveness. In the second study, 4- and 5-year-olds' physiologic reactivity was examined as a predictor of later overt emotional reaction to venipuncture episodes. Children's overt emotional reactions were consistent across repeated venipunctures, and girls were more visibly distressed than boys. As in the first study, physiologic reactivity was generally unrelated to children's behavioral responses. Findings have implications for assumptions about the degree of coupling between biological and behavioral emotional systems in childhood. PMID- 11044864 TI - The impact of stimulus 'value' in infant novelty preference. AB - Various factors have been identified that influence the robust phenomenon of novelty preference in infants. The present study consisted of two experiments that investigated whether conditioned 'value' is another variable that influences infants' preferences. In Experiment 1, novelty preference was established to simple tones despite the tones differing only in frequency. In Experiment 2, novelty preference was manipulated by pairing a primary reinforcer with the familiarization tone such that the conditioned value overrode novelty preference. The findings raise questions about the universality of predicting infants' preferences solely on the foundation of amount of stimulus experience. PMID- 11044865 TI - Temperature asymmetry and behavior. AB - Asymmetries in the surface temperature of the fingertips of the right and left hands were gathered on four independent samples of children ranging in age from 4 to 8 years (N = 398) while watching film clips in order to determine if the direction or magnitude of asymmetry was related to behavioral signs of fear or inhibition. The left index finger was cooler than the right index finger for all four groups. The right ring finger was cooler than the left ring finger for two of the four groups, and of similar temperature for the other two groups. There was no relation between direction of asymmetry and behavior and only a modest relation between a large temperature asymmetry between the index fingers and behavioral signs of very high fear or inhibition. There was no relation between asymmetry and fear or inhibition across all children, suggesting the utility of examining extreme scores. PMID- 11044866 TI - Organizational and activational effects of gonadal steroid hormones on the EEG of male and female rats. AB - To analyze organizational and activational effects of sex steroids on adult rat electroencephalographic activity (recorded at postnatal day 100), seven groups were included: males (48)-intact, neonatally or adult castrated; females (64) intact, ovariectomized and exposed pre- or neonatally to testosterone propionate. In males, neonatal orchidectomy increased beta relative power, whereas both neonatal and adult castration reduced interparietal correlation. In females, prenatal testosterone administration produced higher theta absolute power; theta relative power was higher in all experimental groups, whereas beta1 and beta2 were decreased by prenatal and increased by neonatal virilization; prenatal virilization enhanced, while neonatal virilization and adult ovariectomy decreased interparietal correlation. These data indicate that females are more sensitive to early prenatal than to neonatal organizational effects of sex steroids, and some electroencephalographic features are feminized in castrated males and virilized in perinatally androgenized females. PMID- 11044867 TI - The current state of the journal. PMID- 11044868 TI - What is the prevalence of environmental hazards in the homes of dementia sufferers and are they associated with falls. AB - OBJECTIVES: To examine the frequency of environmental hazards in the homes and care environments of patients with dementia and their associations with falls. METHOD: Falls were prospectively assessed in 65 dementia patients using carer diaries, and the safety of the environment assessed by an occupational therapist using a home hazard checklist. RESULTS: Hazards were found in 20 (95%) of patients' own homes and 31 (74%) of residential or nursing home environments (care environments). Patients' homes had a mean of 5.4 hazards compared to a mean of 1.8 hazards in care environments, with two or more hazards in 90% of patients' homes and 52% of care environments. Common hazards included low chairs, an absence of grab rails (toilet area), toilets too low and a missing second banister on the stairs. There was no significant association between the number of hazards and the number of falls, although 13 (10%) falls could be attributed to a specific hazard. CONCLUSION: Rigorous assessment of the patient's environment revealed multiple rectifiable risks that were contributory to a significant minority of falls. PMID- 11044869 TI - Use of donepezil for the treatment of mild-moderate Alzheimer's disease: an audit of the assessment and treatment of patients in routine clinical practice. AB - There have been a number of randomised, placebo-controlled trials of donepezil in the treatment of mild-moderate Alzheimer's disease and these report significant benefits for a proportion of patients. Little is known about the use of donepezil in routine clinical practice. The aims of this study were to examine the use of donepezil in routine clinical practice and to identify some of the practical and resource implications associated with treatment. A number of areas were examined against published guidelines including assessment, diagnosis, initiation of treatment, monitoring and discontinuation of treatment. This was a retrospective case note study involving patients with mild - moderate Alzheimer's disease over a one-year period. One hundred and seventeen patients were commenced on donepezil and 93 successfully completed three months of treatment. Of these, 47% demonstrated an improvement in cognition, activities of daily living or carer observation (or a combination). Compliance with accepted guidelines with respect to assessment, diagnosis and monitoring requires a standardised approach that has both clinical and resource implications. PMID- 11044870 TI - Development of an indicator to identify inappropriate use of benzodiazepines in elderly medical in-patients. AB - Studies reporting the quantity of benzodiazepines used are purely descriptive and cannot comment on the quality or appropriateness of prescribing benzodiazepines. An indicator to assess the appropriateness of prescribing benzodiazepines was developed from published literature. The applicability of the indicator was discussed in a multidisciplinary forum. The indicator uses clinical data currently available to the prescriber. The indicator, in the form of an algorithm, was applied to assess the appropriateness of prescribing of benzodiazepines to medical in-patients aged < or =65 years at 17 hospitals in England and Wales. Prescribing data were collected on 1391 patients. Appropriateness of prescribing of 311 benzodiazepines were assessed. Benzodiazepines were prescribed appropriately for 110/311 (35%) prescriptions and inappropriately for 201/311 (65%) prescriptions. Initiation of benzodiazepine for no acceptable indication was the commonest reason for inappropriate prescribing. The instrument identifies the appropriateness of prescribing of benzodiazepines and can be utilised by non-physicians. PMID- 11044871 TI - Motor function and disability in the dementias. AB - Epidemiological and neuropathological series have identified three predominant dementing processes; Alzheimer's disease (AD), vascular dementia (VaD) and dementia associated with Lewy bodies (termed Parkinson's disease dementia (PDD) in this paper). While each has its own distinguishing features and by definition all impact upon day to day functioning, no random community derived sample has examined clinical features as defined by gait and balance abnormalities and compared disability ratings of the three dementias simultaneously. Six hundred and forty-seven community dwelling subjects participated in the Sydney Older Persons Study and of these 537 participated in a medical assessment. Of these 537,482 informants rated disability. Gait and balance abnormalities of the three major dementias were identified and the association of the dementias with disability examined. The three major dementias showed evidence of both slowing and ataxia in gait and balance tests. This was maximal in those with PDD. Similarly, all showed evidence of disability that was maximal in those with PDD. In conclusion, this study has identified that gait abnormalities are present in all three dementias to a varying degree. It is hypothesised that the varying levels of disability observed are a consequence of the varying levels of motor impairment, resulting in greater levels of disability in those with PDD. PMID- 11044872 TI - Effects of age and gender on elderly suicide rates in Catholic and Orthodox countries: an inadvertent neglect? AB - When compared to suicide rates in the general population, it may be expected that elderly suicide rates would be lower in Catholic and Orthodox societies than in non-Catholic or non-Orthodox countries because of religious affiliations and extended family traditions. National suicide rates in the general population were compared with rates in the sub-population of those aged over 75 years. Proportionately, there are significantly higher suicide rates in elderly men in Catholic and Orthodox countries, compared to rates in other countries, with a trend for similar findings among women. There may be important implications on health and social policy and clinical practice in the efforts to reduce suicide rates among elderly people. PMID- 11044873 TI - MRI volumetric correlates of white matter lesions in dementia with Lewy bodies and Alzheimer's disease. AB - The aim of the study was to examine the relationship between white matter changes on magnetic resonance imaging (MRI), brain atrophy and ventricular dilation in late-life dementias. T(1)-weighted, T(2)-weighted, and proton density MRI scans were acquired in subjects with Alzheimer's disease (AD, N=25) and dementia with Lewy bodies (DLB, N=27). Total brain and ventricular volumes were measured and white matter lesions rated using a semi-quantitative scale. Periventricular hyperintensities (PVH) were found to independently correlate with advancing age and increasing ventricular dilatation in all subjects. In contrast, deep white matter hyperintensities (DWMH) did not correlate with measures of brain atrophy, ventricular dilatation or age, but were associated with a history of hypertension. These findings support the hypothesis that PVH and DWMH are pathologically diverse and that white matter change in AD and DLB may be determined by similar processes. In particular, PVH appear to be linked to atrophic processes involving ventricular enlargement and DWMH to ischaemic risk factors. PMID- 11044874 TI - A preliminary study of dream-telling among mentally healthy elderly: no adverse effects on life or sleep quality. AB - While there have been several studies about dreams and dreaming among the elderly, there does not seem to have been any study of the effects of regular dream-telling (without interpretation). Listening to dreams could become a regular part of caring for the elderly and infirm. The effects of regular dream telling in mentally healthy elderly clinical research volunteers were measured on several variables using standardized testing and self-report: life satisfaction, intrapsychic boundaries, sleep quality, sleep duration, dream recall, dream tone, and dream epoch, and were compared with two control groups. The six variables showed no significant differences among the three groups, indicating that dream telling produced no adverse effects. The present findings seem to imply that dream-telling is not dangerous for mentally healthy individuals and may thus serve as a baseline for future studies involving geriatric patients with mental disorders or elderly undergoing significant life-events, e.g., bereavement or retirement, using the method of regular dream-telling. PMID- 11044875 TI - The interplay of institution and family caregiving: relations between patient hassles, nursing home hassles and caregivers' burnout. AB - This population-based study investigated the relationship between stresses (hassles) and burnout for 30 family caregivers and their institutionalized demented elderly. The Burnout Measure, the Patient Hassles Scales and the Nursing Home Hassles Scale were used. Hassles included: patient hassles (cognitive, behavior, basic ADL) and nursing home hassles (caregiver - staff, patient - staff, practical/logistical). The caregiver's characteristics are described in relation to burnout and the caregiver's most frequent hassles are discussed. All subscales except basic ADL were correlated to burnout. However, regression analysis showed the nursing home hassles to be the most important stresses explaining variance in burnout among family caregivers. PMID- 11044876 TI - Depressive symptoms predicting six-year mortality in depressed elderly finns. AB - The specific symptoms of depression associated with increased mortality in the depressed elderly are poorly known. The aim of this paper is to analyse the individual depressive symptoms measured by the Zung Self Rated Depression Scale (ZSDS) and the Hamilton Rating Scale for Depression (HRSD) in association with mortality among depressed elderly subjects. The population consisted of 169 depressed (DSM-III criteria) aged (65+ years) persons from a Finnish epidemiological research project. The follow-up for deaths continued for about 6 years. When age, sex, smoking, physical health and functional abilities were taken into account, dissatisfaction, weight loss and gastrointestinal symptoms (anorexia and constipation) predicted mortality together with high age and poor physical health. Weight loss was related to an increased risk of death, specifically in the depressed. Dissatisfaction and gastrointestinal symptoms were more general markers of increased mortality. PMID- 11044877 TI - Dementia research in China. AB - There is little information on dementia research in China in the international literature. This paper is an overview of studies on dementia conducted in China. Studies on dementia research in China were identified through a literature search, as well as through consulting Chinese psychiatrists and neurologists with expertise in the field. These studies were then reviewed. Epidemiological studies on dementia in China find the rates of dementia at the lower end of rates in Caucasians. There seems to be regional variation in the ratio of vascular dementia to Alzheimer's disease in China. In addition to epidemiological studies, there are studies on the genetics, psychopathology, neurophysiology and drug treatment in dementia. There is much research activity in China in the field of dementia and further cross-cultural studies are useful to examine the cultural differences in various areas of dementia research. PMID- 11044878 TI - MRI findings differentiate between late-onset schizophrenia and late-life mood disorder. AB - OBJECTIVES: Compare MRI scans of patients with late-onset schizophrenia, late life depression and late-life bipolar disorder to age- and gender-matched controls. MRI head scans of 14 patients in each diagnostic group and 21 patients in the normal control group were compared. Subjects were recruited from inpatient and outpatient services. MEASURES: The CERAD MRI rating algorithm was used to rate degree of atrophy. RESULTS: Patients with bipolar and unipolar disorder had greater left sylvian fissure and left and right temporal sulcal enlargement, and more bilateral cortical atrophy than normals. Patients with late-onset schizophrenia had larger right temporal horns and larger third ventricles. These findings validate the distinctions between late-life affective disorder and late onset schizophrenia and mirror changes reported in younger individuals. They may reflect underlying structural and functional abnormalities found in neuropathologic and functional imaging studies. PMID- 11044879 TI - Undue concern for others' opinions deters caregivers of impaired elderly from using public services in rural Japan. AB - OBJECTIVE: To determine whether opinions of others may discourage the use of public services for the elderly. DESIGN: Users and non-users were compared regarding several variables including caregivers' concerns about opinions of others. SETTING: Matsuyama Town, located in a rural area of northern Japan. SUBJECTS: Seventy pairs of (1) elderly persons in need of care assessed by the procedure employed by the Ministry of Health and Welfare, and (2) their respective family caregivers. MEASURES: Cognitive function (Hasegawa Dementia Rating Scale), diagnosis of dementia, activities of daily living (ADL: Barthel Index), caregivers' burden (Zarit Caregiver Burden Interview), behavioral disturbances (Dementia Behavioural Disturbance Scale), caregiver interviews. RESULTS: A logistic regression analysis revealed that those who looked after the elderly with severe dependency in ADL were three times more likely to use public services (OR = 3.33, 95% CI = 1.02 - 10.88, p = 0.04). Those concerned about what others think or say were less likely to use public services (OR = 0.22, 95% CI = 0.06 - 0.78, p = 0.01) than those who did not. CONCLUSIONS: Caregivers' undue concern for the opinions of others apparently deterred them from using public services. The overriding notion of care for the elderly as a family duty is still prevalent in rural Japan. It would be useful for the government to launch a public awareness programme to help caregivers understand the benefits of services available for the elderly. PMID- 11044880 TI - Assessment of informal services to demented people with the RUD instrument. PMID- 11044881 TI - The effect of a behaviour rating scale on aggressive behaviour in Dutch nursing homes. PMID- 11044882 TI - Current awareness in geriatric psychiatry [bibliography]. PMID- 11044883 TI - Synaptic plasticity is impaired in rats with a low glutathione content. AB - Long-term potentiation (LTP) is a sustained increase in the efficacy of synaptic transmission, based on functional changes involving pre- and postsynaptic mechanisms, and has been considered a cellular model for learning and memory. The sulphurated tripeptide glutathione acts as a powerful antioxidant agent within the nervous system. Recent in vitro studies suggest that the cellular redox status might influence the mechanisms involved in synaptic plasticity. It is not known, however, how glutathione depletion might affect LTP. In the present study, we evaluated the input-output relationships, LTP, and paired-pulse interactions in rats with low glutathione levels induced by systemic injection of diethylmaleate. Our results in anesthetized rats show that the basic synaptic transmission between the perforant pathway and the dentate gyrus granule cells was not affected by glutathione depletion. However, in the same synapses it was not possible to induce prolonged changes in synaptic efficacy (LTP). Paired-pulse facilitation was also absent in the treated animals, suggesting an impairment of short-term synaptic interactions. These findings indicate that low content of glutathione can impair short-term and long-term mechanisms of synaptic plasticity and stress the importance of the redox balance in the normal function of brain circuitry. PMID- 11044884 TI - Putative role of presynaptic alpha7* nicotinic receptors in nicotine stimulated increases of extracellular levels of glutamate and aspartate in the ventral tegmental area. AB - We have previously provided evidence that the stimulatory action of systemic nicotine on dopamine release in the rat nucleus accumbens is initiated in the ventral tegmental area (VTA), and that it appears to be mediated partly through an indirect, presynaptic mechanism. Thus, it was found that blockade of N-methyl D-aspartate (NMDA) receptors in the VTA attenuates the enhancing effect of nicotine on extracellular levels of dopamine in the nucleus accumbens. Moreover, the nicotine-induced dopamine output in the nucleus accumbens was found to be blocked by pretreatment with methyllycaconitine (MLA) in the VTA, indicating a role for alpha7* nicotinic acetylcholine receptors (nAChRs) in this mechanism. Thus, nicotine may exert its effects in the VTA through stimulation of alpha7* nAChRs localized on excitatory amino acid (EAA)ergic afferents. To test this hypothesis, we here measured extracellular concentrations of glutamate and aspartate in the VTA in response to systemic nicotine, with or without concurrent infusion of MLA in the VTA, using microdialysis in anaesthetized rats. Since the medial prefrontal cortex is an important source of EAA input to the VTA, we also assessed the density of alpha-bungarotoxin binding sites in the VTA in rats lesioned bilaterally in the prefrontal cortex with ibotenic acid and in sham lesioned rats by means of quantitative autoradiography. Nicotine (0.5 mg/kg, s.c.) significantly increased extracellular levels of both aspartate and glutamate in the VTA. MLA (0.3 mM) infused locally in the VTA prevented the nicotine-induced increase in glutamate and aspartate levels. Ibotenic acid lesions of the prefrontal cortex decreased the density of alpha-bungarotoxin binding sites in the VTA by about 30%. These data indicate that nicotine increases the extracellular levels of excitatory amino acids in the VTA through stimulation of nAChRs in the VTA and that part of the alpha7* nAChR population in the VTA is localized on neurons originating in the prefrontal cortex. PMID- 11044885 TI - Do synaptic rearrangements underlie compensatory reflex enhancement in spinal motoneurons after partial cell loss? AB - In adult cats, avulsion of a spinal ventral root induces retrograde cell death among the corresponding motoneurons and, also, enhanced monosynaptic reflexes ipsilaterally in the adjacent uninjured spinal cord segments. The present study investigates possible mechanisms behind this reflex potentiation. At 1-12 weeks after unilateral L7 ventral root avulsion, the L7 dorsal root ganglia were bilaterally injected with choleragenoid-HRP to light microscopically quantify the amount of HRP-labeled terminals in the motor nuclei of the lesioned L7 segment and adjacent intact L6+S1 segments. In addition, motoneuron synaptology and individual HRP-labeled boutons were analyzed electron microscopically. In the L7 segment, the loss of motoneurons at 12 weeks after ventral root avulsion was accompanied by a marked loss of HRP-labeled boutons in the corresponding ventral horn. In the L6/S1 segments, the monosynaptic reflex enhancement found ipsilaterally at 12 weeks postoperatively (mean 212%) was not accompanied by an increased HRP-labeling in the ventral horn (mean 109%), indicating that no sprouting or enlargement of the monosynaptic boutons had occurred. Ultrastructurally, the values for apposition length, total active site length, cross-sectional area, and mitochondrial density of the labeled boutons were also similar between the two sides. However, ipsilaterally the L6/S1 motoneurons exhibited an increased membrane covering by presumably excitatory boutons. The present results indicate that after partial cell death in a motoneuron pool the remaining motoneurons may undergo compensatory synaptic rearrangements leading to increased excitability and enhanced reflexes. PMID- 11044886 TI - Expression and distribution of CYP2C enzymes in rat basal ganglia. AB - The function and integrity of the basal ganglia is modulated by sex steroids whose activity may be controlled by P450 enzymes, such as members of the CYP2C subfamily. The expression of CYP2C enzymes in rat basal ganglia was examined by immunohistochemistry along with some of the factors that might control their expression. Whereas no CYP2C11 or CYP2C12 immunoreactivity was detected in the basal ganglia of either male or female rats, marked CYP2C13 immunoreactivity was evident in neurones of the subthalamic nucleus, substantia nigra, and interpeduncular nucleus. Strong CYP2C13 immunoreactivity was also expressed in the cortex, olfactory tubercle, hippocampus, dentate gyrus, hypothalamic nuclei, medial habenular nucleus, red nucleus, and medial forebrain bundle. Similar results were found in male and female rats. Following 6-hydroxydopamine lesioning of the nigro-striatal tract, tyrosine hydroxylase immunoreactivity was absent and CYP2C13 immunoreactivity was decreased markedly in the substantia nigra pars compacta, implying its presence in dopaminergic neurones. Modulation of sex steroids, using castrated rats, had no effect on the number of CYP2C13 positive neurones in the substantia nigra pars compacta. These results indicate that CYP2C13 protein is constitutively and widely expressed in rat brain. However, its expression is not sex-specific and is unaffected by castration. The role of CYP2C13 in brain is unknown but it may be involved in the generation of neurosteroids and catecholoestrogens. PMID- 11044887 TI - Selective in vitro and in vivo binding of [(125)I]ADAM to serotonin transporters in rat brain. AB - An improved iodinated tracer, ADAM (2-((2-((dimethylamino)methyl)- phenyl)thio)-5 iodophenylamine) for imaging serotonin transporters (SERT) with single photon emission computerized tomography (SPECT), was prepared and characterized. Scatchard analysis of saturation binding of [(125)I]ADAM to rat frontal cortical membrane homogenates gave a K(d) value of 0.15 +/- 0.03 nM and a B(max) value of 194 +/- 65 fmol/mg protein. Biodistribution of [(125)I]ADAM in rat brain after an iv injection showed a high specific binding in the regions of hypothalamus, cortex, striatum, and hippocampus, where SERT are concentrated and the specific binding peaked at 120-240 min postinjection [(hypothalamus-cerebellum)/cerebellum = 4.3 at 120 min post-iv injection]. Moreover, the specific hypothalamic uptake was blocked by pretreatment with SERT selective competing drugs, such as paroxetine and (+)McN5652, while other noncompeting drugs, such as ketanserin, raclopride, and methylphenidate, showed no effect. The radioactive material recovered from rat brain homogenates at 120 min after [(125)I]ADAM injection showed primarily the original compound (>90%), a good indication of in vivo stability in the brain tissues. Both male and female rats showed similar and comparable organ distribution pattern and regional brain uptakes. Ex vivo autoradiograms of rat brain sections (120 min after iv injection of [(125)I]ADAM) showed intense labeling in several regions (olfactory tubercle, lateral septal nucleus, hypothalamic and thalamic nuclei, globus pallidus, central gray, superior colliculus, substantia nigra, interpeduncular nucleus, dorsal and median raphes, and locus coerulus), which parallel known SERT density. These results strongly suggest that the novel tracer ADAM is superior to the congers (i.e., IDAM) reported previously. When labeled with I-123, ADAM will be an improved and useful SPECT imaging agent for SERT in the brain. PMID- 11044888 TI - Localization of dopamine D1 and D2 receptors in the rat neostriatum: synaptic interaction with glutamate- and GABA-containing axonal terminals. AB - In order to determine the synaptic interactions between the glutamate- and GABA containing axonal terminals and the two subpopulations of medium spiny neurons in the rat neostriatum, double immunocytochemistry was performed. Sections of perfuse-fixed rats were used. Immunoreactivity for dopamine D1 and D2 receptors was used as a marker for the two subpopulations of spiny neurons that give rise to the direct and indirect pathways, respectively. Receptor immunoreactivity was first revealed by preembedding immunostaining. Postembedding colloidal gold labeling was then performed to reveal immunoreactivity for glutamate or GABA. The results were analyzed at the electron microscopic level. Both the D1 immunoreactive, presumed striatonigral/entopeduncular neurons, and the D2 immunoreactive, presumed striatopallidal neurons, were found to receive qualitatively similar synaptic inputs from glutamate-immunoreactive terminals and GABA-immunoreactive terminals. The present results indicate that the different classes of spiny neurons are thus likely to be under a similar regime of excitatory and inhibitory control. PMID- 11044889 TI - Autoradiographic localization of 5-HT(2A) receptors in the human brain using [(3)H]M100907 and [(11)C]M100907. AB - M100907 (MDL 100907, R-(+)-alpha-(2, 3-dimethoxyphenyl)-1-[2-(4 fluorophenyl)ethyl]-4-piperidinemethanol++ +) is a new selective antagonist of 5 HT(2A) receptors. The compound has been labeled with (11)C and proved useful for in vivo studies of 5-HT(2A) receptors using positron emission tomography (PET). In the present study the distribution of 5-HT(2A) receptors was examined in the postmortem human brain using whole hemisphere autoradiography and [(3)H]M100907 and [(11)C]M100907. The autoradiograms showed very dense binding to all neocortical regions, whereas the hippocampus was only weakly labeled with [(3)H]M100907. Other central brain regions, such as the basal ganglia and thalamus, showed low [(3)H]M100907 binding, reflecting low densities of 5-HT(2A) receptors. The cerebellum or structures of the brain stem were virtually devoid of 5-HT(2A) receptors. [(11)C]M100907 gave images qualitatively similar to those of [(3)H]M100907, although with lower spatial resolution. The labeling of human 5 HT(2A) receptors with [(3)H]M100907 was inhibited by the addition of the 5-HT(2A) receptor blockers ketanserin or SCH 23390 (10 microM), leaving a very low background of nonspecific binding. The 5-HT(1A) receptor antagonist WAY-100635 and the D(2)-dopamine receptor antagonist raclopride had no effect on the binding of [(3)H]M100907. The selective labeling of 5-HT(2A) receptors with [(3)H]M100907 clearly shows that this compound is suitable for further studies of the human 5 HT(2A) receptor subtype in vitro. The in vitro autoradiography of the distribution of 5-HT(2A) receptors obtained with radiolabeled M100907 provides detailed qualitative and quantitative information on the distribution of 5-HT(2A) receptors in the human brain as well as reference information for the interpretation of previous initial results at much lower resolution in humans in vivo with PET and [(11)C]M100907. PMID- 11044890 TI - Synergistic interactions between nicotine and cocaine or methylphenidate depend on the dose of dopamine transporter inhibitor. AB - There is a greater prevalence of cigarette smoking among cocaine-dependent individuals and hyperactive children treated with stimulants (e.g., methylphenidate, MP). However, little is known about the neurochemical basis of the interaction between nicotine and cocaine or MP. It is thought that the reinforcing effects of cocaine and MP are due partly to increases in synaptic DA in the nucleus accumbens (NAc). These measurable increases are secondary to the blockade of the DA transporter. In contrast, nicotine stimulates acetylcholine receptors located presynaptically on dopaminergic projections from the ventral tegmental area (VTA) to the NAc and increases DA transmission. Here we investigate the effects of nicotine on NAc DA in animals simultaneously injected with cocaine or MP. Coadministration of nicotine (0.4 mg/kg s.c.) and cocaine (10 mg/kg i.p.) or MP (5 mg/kg i.p.) increased the extracellular NAc DA levels in an additive manner, while coadministration of nicotine (0. 4 mg/kg s.c.) and a higher dose of cocaine (20 mg/kg) or MP (10 mg/kg) clearly produced a synergistic elevation in NAc DA. These findings suggest that the degree of DA transporter (DAT) occupancy contributes to the synergistic interaction between nicotine and cocaine or MP. PMID- 11044891 TI - Characterization of (125)I-IABN, a novel azabicyclononane benzamide selective for D2-like dopamine receptors. AB - The properties of an (125)I-labeled structural analog of 2, 3-dimethoxy-N-[9-(4 fluorobenzyl)-9-azabicyclo[3.3. 1]nonan-3beta-yl]benzamide (MABN), (125)I-IABN, are described. (125)I-IABN was developed as a high-affinity radioligand selective for the D2-like (D2, D3, and D4) dopamine receptor subtypes. (125)I-IABN binds with picomolar affinity and nonselectively to rat D2 and D3 dopamine receptors expressed in Sf9 and HEK 293 cells. (125)I-IABN binds with 7- to 25-fold lower affinity to human D4.4 dopamine receptors expressed in HEK 293 cells. Dissociation constants (Kd) calculated from kinetic experiments were in agreement with equilibrium Kd values obtained from saturation binding studies. Saturation plots of the binding of (125)I-IABN with rat caudate membrane preparations were monophasic and exhibited low nonspecific binding. The pharmacologic profile of the binding of (125)I-IABN to rat caudate was consistent with a D2-like receptor, suggesting that the ligand binds primarily to D2 dopamine receptors. In addition, IABN was found to bind with low affinity to D1 dopamine receptors, as well as to the sigma1 and sigma2 receptor subtypes. Quantitative autoradiographic studies using rat brain slices indicate that (125)I-IABN selectively labels the striatum and the olfactory tubercle area, which is consistent with the labeling of D2-like receptors. IABN blocks dopamine-dependent inhibition of adenylyl cyclase activity at D2 or D4.4 receptors expressed in HEK cells. Therefore, (125)I-IABN appears to be a high-affinity, selective antagonist at D2-like dopamine receptors. Finally, a unique property of the azabicyclononane benzamide (125)I-IABN compared to previously studied substituted benzamides is that the binding of this radioligand is not effected by variations in Na(+) concentration. PMID- 11044892 TI - Insulin-like growth factor-I and its receptor in the frontal cortex, hippocampus, and cerebellum of normal human and alzheimer disease brains. AB - Assimilated evidence indicates that the neurotoxic potential of amyloid beta (Abeta) peptide and an alteration in the level of growth factor(s) may possibly be involved in the loss of neurons observed in the brain of patients suffering from Alzheimer disease (AD), the prevalent cause of dementia in the elderly. In the present study, using receptor binding assays and immunocytochemistry, we evaluated the pharmacological profile of insulin-like growth factor-I (IGF-I) receptors and the distribution of IGF-I immunoreactivity in the frontal cortex, hippocampus, and cerebellum of AD and age-matched control brains. In control brains, [(125)I]IGF-I binding was inhibited more potently by IGF-I than by Des(1 3)IGF-I, IGF-II or insulin. The IC(50) values for IGF-I in the frontal cortex, hippocampus, and cerebellum of the normal brain did not differ significantly from the corresponding regions of the AD brain. Additionally, neither K(D) nor B(max) values were found to differ in the hippocampus of AD brains from the controls. At the regional levels, [(125)I]IGF-I binding sites in the AD brain also remained unaltered compared to the controls. As for the peptide itself, IGF-I immunoreactivity, in normal control brains, was evident primarily in a subpopulation of astrocytes in the frontal cortex and hippocampus, and in certain Purkinje cells of the cerebellum. In AD brains, a subset of Abeta-containing neuritic plaques, apart from astrocytes, exhibit IGF-I immunoreactivity. These results, taken together, suggest a role for IGF-I in compensatory plasticity and/or survival of the susceptible neurons in AD brains. PMID- 11044893 TI - In vivo microdialysis assessment of extracellular serotonin and dopamine levels in awake monkeys during sustained fluoxetine administration. AB - Fluoxetine (FLU) rapidly enhances extracellular (EC) serotonin (5-HT) in rodent brain, whereas the antidepressant effects of this drug in humans are typically not observed for 2-3 weeks. Thus, the effects of chronic oral FLU administration on neocortical and hippocampal EC 5-HT, and on caudate EC 5-HT and dopamine (DA), were examined in awake monkeys (Macaca fascicularis) using in vivo microdialysis (10.0 mg/kg; 3, 7, 14, and 21 days). On day 3, 5-HT was significantly increased above baseline levels in hippocampus (HC) and caudate. There was a trend for an increase in neocortex EC 5-HT levels. However, by day 7 5-HT remained significantly elevated only in HC, although 5-HT levels elsewhere had not completely returned to baseline. In contrast, levels of the 5-HT metabolite, 5 HIAA, were significantly reduced in all brain regions at all time points. Caudate DA levels tended to be decreased throughout FLU treatment. Local FLU and K(+) infusion were also used at various times during chronic systemic FLU administration to evaluate changes in functional synaptic regulation. In general, these results, along with the significant decrease in 5-HIAA levels and the tendency for basal EC 5-HT levels to remain modestly elevated only in HC during sustained FLU administration, suggest a reduction in releasable pools of 5-HT. Taken together with the trend for a decrease in caudate EC DA levels, these results do not appear to support the current hypothesis regarding the mechanism of action of SSRI antidepressants-that monoaminergic neurotransmission is progressively augmented during chronic treatment. PMID- 11044894 TI - Fluoxetine increases the anorectic and long-term dopamine-depleting effects of phentermine. AB - The anorectic drug phentermine produces dose-related toxic effects on brain dopamine (DA) neurons in animals. Until recently, phentermine was widely used in combination with fenfluramine for purposes of appetite suppression and weight loss. With the recent withdrawal of fenfluramine from the market, many people have begun combining phentermine with fluoxetine, a serotonin reuptake inhibitor which also produces mild anorectic effects. Fluoxetine, in addition to inhibiting serotonin reuptake, inhibits hepatic mixed function oxidase, which plays an important role in the metabolic degradation of amphetamines. The purpose of the present study was to assess the effects of fluoxetine on the anorectic and DA neurotoxic effects of phentermine in mice. Phentermine, in combination with fluoxetine, produced greater reductions in food intake and body weight than phentermine alone. The phentermine/fluoxetine combination also produced greater long-term reductions in brain DA levels than phentermine alone, likely reflecting greater DA neurotoxicity of the drug combination. Brain concentrations of phentermine were also found to be higher in animals pretreated with fluoxetine. These findings indicate that fluoxetine potentiates both the anorectic and DA neurotoxic effects of phentermine, probably by increasing phentermine brain levels. The clinical significance of these findings remains to be ascertained. PMID- 11044895 TI - Locomotor activity and occupancy of brain cannabinoid CB1 receptors by the antagonist/inverse agonist AM281. AB - The goals of this study were to examine the relationship between intravenous doses of the cannabinoid CB1 receptor antagonist AM281 (N-(morpholin-4-yl)-5-(4 iodophenyl)-1-(2, 4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide) and the degree of occupancy of this receptor, and to relate occupancy to the ability of this compound to antagonize the sedative effects of the cannabinoid receptor agonist WIN 55,212-2. Occupancy was determined by measuring the ability of intravenous doses of AM281 to inhibit in vivo binding of [(131)I]AM281 in brain areas, and locomotor activity was assessed by measuring the rate of beam crossings in a photocell apparatus. As previously documented, WIN 55,212-2 (1 mg/kg, i.v.) significantly reduced locomotor activity at early times after administration. Co-injection of AM281 (0.3 mg/kg i/v) and WIN 55, 212-2 restored the rate of beam crossings to that seen on injection of vehicle. In addition, AM281 (0.3 mg/kg i/v) approximately doubled locomotor activity between 60-120 min when injected alone. The IC(50) value for displacement of [(131)I]AM281 by AM281 was 0.45 mg/kg. These observations confirm earlier indications that AM281 is a CB1 receptor antagonist or inverse agonist and suggest the existence of an endogenous cannabinoid tone that moderates exploratory locomotor activity. PMID- 11044896 TI - Sensitivity of binding of high-affinity dopamine receptor radioligands to increased synaptic dopamine. AB - PET and SPECT studies have documented that D2 radioligands of moderate affinity, but not radioligands of high affinity, are sensitive to pharmacological challenges that alter synaptic dopamine levels. The objective of this work was to determine whether the brain kinetics of high-affinity radioligands for dopamine D1 ([(3)H]SCH 23390) and D2 ([(123)I]epidepride) receptors were altered by a prolonged elevation of synaptic dopamine induced by the potent cocaine analog RTI 55. Mice were injected intravenously with radioligands either 30 min after or 4 h before intraperitoneal administration of RTI-55 (2 mg/kg). In separate experiments, the pharmacological effects of RTI-55 were assessed biochemically by measuring uptake of dopamine in synaptosomes prepared from RTI-treated mice and behaviorally by monitoring locomotor activity. Consistent with the expected elevation of synaptic dopamine, RTI-55 induced a long-lasting decrement in dopamine uptake measured ex vivo, and a prolonged increase in locomotor activity. RTI-55 injected prior to the radioligands induced a significant (P < 0.05) increase in striatal concentration of [(123)I]epidepride at 15 min, relative to saline-treated controls, but there were no differences between the two groups at later time-points. For [(3)H]SCH 23390, both initial striatal uptake and subsequent clearance were slightly increased by preadministration of RTI-55. Administration of RTI-55 4 h after the radioligands (i.e., when it was presumed that a state of near equilibrium binding of the radioligands had been reached), was associated with a significant reduction of striatal radioactivity for both radiotracers. Our results are consistent with increased competition between dopamine and radioligand for binding to both D1 and D2 receptors after treatment with RTI-55. We suggest that the magnitude of the competition is reduced by failure of the receptor binding of high-affinity radioligands to rapidly attain equilibrium. PMID- 11044897 TI - Functional downregulation of GluR2 in piriform cortex of kindled animals. AB - We have previously shown kindling-induced downregulation of the AMPA receptor GluR2 subunit in piriform cortex, as measured by Western blotting. In the present studies, we performed whole-cell patch clamp analysis of AMPA receptor-mediated currents from kindled and control animals to determine if the downregulation observed previously had any functional significance. These experiments were done in the absence and presence of N-hydroxyphenylpropanoyl spermine (HPPS), a polyamine that blocks currents through AMPA receptors lacking GluR2. We report that AMPA receptor-mediated currents recorded from piriform cortex layer II pyramidal cells in slices from animals kindled to 10 fully generalized seizures were blocked by HPPS. In contrast, application of HPPS had no effect on current amplitude in control animals, or in animals that had not been fully kindled. Western blotting revealed that decreases in GluR2 were seen in animals that had experienced at least one fully generalized seizure, but were not observed at earlier stages of kindling development. The increased polyamine sensitivity of AMPA receptor-mediated currents in kindled animals is consistent with the hypothesis that kindling induces formation of AMPA receptors that lack GluR2 in piriform cortex pyramidal cells. It has been demonstrated that polyamine sensitivity is directly correlated with the calcium permeability of the AMPA receptor, suggesting that kindling results in the formation of AMPA receptors that are calcium-permeable. Increases in intracellular calcium through these receptors could act as a second messenger and play a role in the initiation of long-term changes that contribute to the pathogenesis of kindling-induced epilepsy. PMID- 11044898 TI - Postsynaptic actin filaments at the giant mossy fiber-unipolar brush cell synapse. AB - The unipolar brush cell (UBC), a small interneuron occurring at high density in the granular layer of the mammalian vestibulocerebellum, receives a giant glutamatergic synapse from a single mossy fiber (MF) rosette, usually on a brush of dendritic branchlets. MF stimulation produces a current in the UBC several orders of magnitude greater in duration than at other glutamatergic synapses. We assumed that the cytoskeleton would have a special role in plasticity of the MF UBC synapse. Neurofilaments and microtubules are enriched in the UBC somatodendritic compartment but are conspicuously absent in close proximity to the giant synapse, where standard electron microscopy reveals a granulo flocculent material. Because osmium tetroxide fixation during sample preparation for standard electron microscopy destabilizes actin filaments, we hypothesized that this subsynaptic granulo-flocculent material is actin-based. After actin stabilization, we observed prominent, but loosely organized, bundles of microfilaments at the subsynaptic region of the MF-UBC synapse that linked the postsynaptic density with the cytoskeletal core of the dendritic branchlets. Confocal fluorescence microscopy and pre- and postembedding immunogold labeling with phalloidin and actin antibodies showed that these microfilaments consist of f-actin and contain little beta-actin. This extraordinary postsynaptic actin apparatus is ideally situated to form a dynamic framework for glutamate receptors and other postsynaptic molecules, and to mediate activity-dependent plastic rearrangements of the giant synapse. PMID- 11044899 TI - Micronucleated erythrocyte frequency in peripheral blood of B6C3F(1) mice from short-term, prechronic, and chronic studies of the NTP carcinogenesis bioassay program. AB - The mouse peripheral blood micronucleus (MN) test was performed on samples collected from 20 short-term, 67 subchronic, and 5 chronic toxicity and carcinogenicity studies conducted by the National Toxicology Program (NTP). Data are presented for studies not previously published. Aspects of protocol that distinguish this test from conventional short-term bone marrow MN tests are duration of exposure, and absence of repeat tests and concurrent positive controls. Furthermore, in contrast to short-term bone marrow MN tests where scoring is limited to polychromatic erythrocytes (PCE), longer term studies using peripheral blood may evaluate MN in both, or either, the normochromatic (NCE) or PCE populations. The incidence of MN-PCE provides an index of damage induced within 72 hr of sampling, whereas the incidence of MN in the NCE population at steady state provides an index of average damage during the 30-day period preceding sampling. The mouse peripheral blood MN test has been proposed as a useful adjunct to rodent toxicity tests and has been effectively incorporated as a routine part of overall toxicity testing by the NTP. Data derived from peripheral blood MN analyses of dosed animals provide a useful indication of the in vivo potential for induced genetic damage and supply an important piece of evidence to be considered in the overall assessment of toxicity and health risk of a particular chemical. Although results indicate that the test has low sensitivity for prediction of carcinogenicity, a convincingly positive result in this assay appears to be highly predictive of rodent carcinogenicity. PMID- 11044900 TI - Interference of tannic acid on the genotoxicity of mitomycin C, methylmethanesulfonate, and nitrogen mustard in somatic cells of Drosophila melanogaster. AB - The modulating effects of tannic acid (TA) on somatic mutation and mitotic recombination induced by methylmethanesulfonate (MMS), nitrogen mustard (HN2), and mitomycin C (MMC) were evaluated in the standard (ST) cross of the wing spot test in Drosophila melanogaster using co- and posttreatment protocols. It was shown that TA alone did not modify the spontaneous frequencies of single and twin spots, which means that this polyphenol neither acts as a genotoxin nor exerts any antigenotoxic effect over spontaneous DNA lesions. However, the simultaneous administration of genotoxins with TA can lead to considerable alterations of the frequencies of induced wing spots in comparison to those with administration of the genotoxins alone. In fact, TA produced a significant increase in HN2-induced wing spots with enhancements between 90 and 160%. For MMS, the enhancement was 38% in the highest TA concentration tested. In contrast, a significant protective action of this polyphenol was observed in combined treatments with MMC (64 to 99% inhibition). Moreover, the data from TA posttreatments demonstrated that this agent is not effective in exerting protective or enhancing effects on the genotoxicity of MMS, HN2, or MMC. One feasible mechanism of TA action is its interaction with the enzyme systems catalyzing the metabolic detoxification of MMS and HN2, which may also be involved in the bioactivation of MMC. PMID- 11044901 TI - Identification of genes responsive to BPDE treatment in HeLa cells using cDNA expression assays. AB - Genotoxic stresses induce cellular responses that can be observed at the level of gene expression. We have studied changes in gene expression following BPDE exposure in HeLa cells by using a cDNA expression array of 597 human genes. After a 53-hr exposure to 0.4 microM BPDE, nine genes were upregulated. The protein products of these genes are: fos-related antigen 2, apoptotic cysteine protease MCH4, DB1 (zinc finger protein 91), transcription factor ETR103, integrin alpha, interleukin-4, interleukin-6, 23-kDa highly basic protein, and ribosomal protein S9. We observed the downregulation of gene expression of three genes: heat-shock protein 27, DNA-binding protein TAX, and NADH-ubiquinone oxidoreductase B18 subunit. These results suggest unknown functions or regulatory circuits for several of the responsive genes and demonstrate the complexity of cellular responses to genotoxic insults. PMID- 11044902 TI - Effect of five dietary antimutagens on the genotoxicity of six mutagens in the microscreen prophage-induction assay. AB - Dietary antimutagens have been studied extensively in the last two decades, using mainly bacterial and mammalian cells. These studies have shown that certain dietary antimutagens, acting individually or as mixtures, are useful in counteracting the effects of certain mutagens and/or carcinogens to which humans are commonly exposed. However, there are some inconsistencies among publications using different bioassays. The general purpose of the research presented here was to conduct a comparative study of the antigenotoxic activity of five dietary antimutagens against six mutagens, using three rather different short-term tests: the Microscreen prophage-induction assay, the Tradescantia micronucleus test, and the Salmonella/mammalian microsome test. In this study I report the results with the Microscreen prophage-induction assay. The antimutagens selected were chlorophyllin, beta-carotene, and vitamins A, C, and E. The mutagens selected were 2-aminoanthracene, benzo[a]pyrene, 2-nitrofluorene, toxaphene, dichlorvos, and nitrofen. The results show that chlorophyllin and beta-carotene inhibited the genotoxicity of all six mutagens; vitamin E inhibited all except dichlorvos; and vitamins C and A inhibited 2-aminoanthracene, benzo[a]pyrene, 2-nitrofluorene, and nitrofen. PMID- 11044903 TI - N-acetyl transferase-2 and bladder cancer risk: a meta-analysis. AB - Interindividual differences in bladder cancer susceptibility may be partly mediated through polymorphic variability in the metabolism of carcinogens. N acetyl transferase-2 (NAT2) has been extensively studied as a risk factor in this context, but the results are inconsistent. In some studies the failure to demonstrate a relationship may be a consequence of a lack of statistical power. To overcome lack of power, data from 21 published case-control studies were pooled in a meta-analysis using a random-effects model. The pooled odds ratio of bladder cancer associated with slow acetylator status was 1.31 (95% CI: 1.11 1.55). The results suggest that NAT2 slow acetylator status is associated with a modest increase in risk of bladder cancer. There was, however, heterogeneity between studies. It is clear from this overview that greater attention should be paid to the design of these types of study. PMID- 11044904 TI - Screening a human population sample for DNA repair gene deficiencies utilizing the protein truncation test. AB - A significant fraction of human cancers are thought to have a genetic component and several lines of evidence suggest that deficiencies in DNA repair may be a contributing factor. Little is known, however, about the frequency and distribution of variants of DNA repair genes in the general human population. The protein truncation test (PTT) was used to screen 136 healthy volunteers for protein-truncating variants of 10 DNA repair genes: APE, CDK7, ERCC1, WAF1, HOGG1, MGMT, POLB, UNG, HAAG, and CCNH. This sample consisted of 41males (30%) and 95 females (70%) with an average age of 25.3 years, ranging from 17 to 60 years of age. No truncating mutations were found in the 10 genes examined in any of the subjects. The 95% confidence interval for a proportion of 0 over the 272 alleles examined per locus is 0-0.01. The calculated frequency of truncating mutations in each of these genes, among the general population, is thus less than 1%. Among the 10 genes tested in 136 people, a single sample had no PCR product for HAAG, even though PCR products were obtained on all other genes. Total RNA dot hybridization confirmed the presence of HAAG mRNA transcripts in this sample. Despite identification of this single DNA repair variant, these results indicate a low frequency of truncating mutations in DNA repair genes in the general population. PMID- 11044905 TI - Mutant frequency and molecular analysis of in vivo lacI mutations in the bone marrow of Big Blue rats treated with 7, 12-dimethylbenz[a]anthracene. AB - Recently, we evaluated lacI mutations in lymphocytes and mammary tissue of Big Blue (BB) rats exposed to 7, 12-dimethylbenz[a]anthracene (DMBA). The results on the time course of mutant induction suggested that the lacI gene may manifest a tissue-specific increase in mutant frequency (MF). To test whether a tissue specific increase in lacI MF is dependent on the cell proliferation rate of a tissue, we examined rapidly proliferating bone marrow cells for DMBA-induced lacI mutations. Seven-week-old female BB rats were given single doses of 0, 20, and 130 mg/kg DMBA by gavage and the lacI MFs in the bone marrow were measured over a period of 14 weeks following treatment. Bone marrow cells had a remarkably low average background MF (3.1 +/- 1.6 x 10(-6) plaque-forming units) and the DMBA induced lacI MFs were significantly higher than control MFs for both doses and at all time points (P < 0.01). The lacI MF in the bone marrow increased for 2 weeks and then remained relatively constant; 20 and 130 mg/kg DMBA produced 34- and 106 fold increases in MF over control MF. DNA sequencing revealed that the majority of DMBA-induced lacI mutations were base-pair substitutions and that A:T --> T:A (48%) and G:C --> T:A (24%) transversions were the predominant types. Thus, the different lacI mutation fixation times observed for bone marrow (2 weeks), mammary (10 weeks), and lymphocytes (6 weeks) suggest that the lacI gene manifests a tissue-specific mutation fixation time, which may depend on the cell proliferation rate of a tissue. In addition, the relatively low spontaneous MF in bone marrow compared with that in other tissues may be useful for increasing the sensitivity of the assay for detecting induced MFs in BB rats. PMID- 11044906 TI - Effects of air pollution and smoking on DNA damage of human lymphocytes. AB - The comet assay is a useful technique for the study of genetic damage in humans exposed to environmental mutagens and carcinogens. In this study the effects of hydrogen peroxide (H(2)O(2)) and ultraviolet (UV) irradiation on 80 healthy individuals living in urban and rural areas with different smoking habits were investigated. Endonuclease III (endo III) treatment was also used to reveal the level of oxidized pyrimidine formation in these groups. The extent of damage and subsequent repair appear to be influenced by the living conditions (urban or rural areas). Smoking, however, was shown to have the most significant effect on DNA damage on all groups studied. PMID- 11044907 TI - Erythrocyte and spermatid micronucleus analyses in mice chronically exposed to potassium bromate in drinking water. PMID- 11044908 TI - Progress in the use of gene transfer methods to treat genetic blood diseases. AB - A report by French physician-scientists suggests a successful application of gene transfer methods in the treatment of two children with severe combined immunodeficiency (SCID) due to defective interleukin 2 receptor common gamma chain. The protocol used in this clinical trial was derived from a number of preclinical and basic studies leading to improved transduction of hematopoietic stem and primitive progenitor cells using retrovirus vectors. These improvements have also been shown to impact transduction of a long-lived progenitor cell in a chemotherapy protocol in cancer patients. The improved results of these human trials come during a period of increased scrutiny and criticism of human gene therapy trials, due, in part, to significant toxicities in some trials using adenovirus-based vectors. The potential efficacy versus toxicity of phase I trials of human gene therapy is also under question. After many years of research, however, there appears to be real evidence that genetic diseases may be successfully treated by gene transfer techniques. Future clinical studies should be based on continued progress in the understanding of the toxicology of gene delivery systems, vector technology, and target cell manipulation. PMID- 11044909 TI - Recombinant adeno-associated virus vector-based gene transfer for defects in oxidative metabolism. AB - Defects in oxidative metabolism may be caused by mutations either in nuclear genes or in mitochondrial DNA (mtDNA). We tested the hypothesis that recombinant adeno-associated virus (rAAV) could be used to complement mtDNA mutations. AAV vector constructs were designed to express the reporter gene encoding green fluorescent protein (GFP), fused to a targeting presequence that directed GFP to be translocated into mitochondria. These vectors mediated expression of mitochondrial-localized GFP, as indicated by fluorescence microscopy and electron microscopy, in respiring human embryonic kidney 293 cells and nonrespiring mtDNA deficient (rho 0) cells. However, when sequences encoding hydrophobic segments of proteins normally encoded by mtDNA were inserted between the presequence and GFP, mitochondrial import failed to occur. In similar experiments, a fusion was created between pyruvate dehydrogenase (PDH) E1 alpha subunit, a nuclear-encoded mitochondrial gene with its own targeting presequence, and GFP. With this construct, expression of GFP was observed in mitochondria in vitro and in vivo. We conclude that the hydrophobicity of mtDNA-encoded proteins limits their ability to be transported from the cytoplasm. However, rAAV-based gene therapy may hold promise for gene therapy of PDH deficiency, the most common biochemically proven cause of congenital lactic acidosis. PMID- 11044910 TI - Purification of recombinant adeno-associated virus vectors by column chromatography and its performance in vivo. AB - Recombinant adeno-associated virus (AAV) holds much promise for human gene therapy. While evidence indicates that AAV mediates long-term gene transfer in several different tissues, difficulty in preparing and purifying this viral vector in large quantities remains a major obstacle for evaluating AAV vectors in clinical trials. The current method of purification, based on sedimentation through cesium chloride, is not scaleable and yields product of insufficient quality. In this article we report a new technique for purifying AAV, using a fully closed two-column chromatography system. Yields of AAV vectors purified by this method are high, potency is increased, and the purity of column-purified preparations is substantially improved. We previously reported a novel method to generate AAV based on an AAV Rep/Cap-containing cell line (B50) and an Ad-AAV hybrid virus, which is amenable to scale-up in bioreactors. By combining the new, fully scaleable purification process we report here with the B50/hybrid production method, it would be feasible to prepare AAV vectors to the scale and purity required for clinical and potential commercial applications. PMID- 11044911 TI - Packaging cell line characteristics and optimizing retroviral vector titer: the National Gene Vector Laboratory experience. AB - During the production of clinical-grade retroviral vector supernatant, we noted significant differences in the lactate production and glucose consumption of various producer cell lines submitted to the National Gene Vector Laboratory (NGVL). Since differences in growth characteristics could be important in determining the optimal culture conditions for maximizing titer, we studied the growth characteristics of three commonly used packaging cell lines: PA317, PG13 and GP+envAM12. A transformed phenotype, assessed by the ability to form colonies in semisolid media, was evident in all three packaging cell lines tested. In confluent cultures, the rates of glucose consumption and lactate production (per cell per hour) were similar for the three lines tested, but the growth rate and culture density varied. PA317 and PG13 continued to expand after reaching confluence, resulting in higher cell densities and subsequent rapid depletion of glucose within the 24-hr observation period. When the cell lines were evaluated for titer optimization, the slower growing packaging cell line GP+envAM12 generally provided the highest titer after 8 hr of culture in confluent roller bottles, while most vectors introduced into PA317 and PG13 cells yielded optimal titers after 24 hr of culture. We also found that the improved titers obtained by culturing cells at 32 degrees C previously reported for PA317 cells do not apply to other packaging cell lines. In particular, PG13 rapidly lost titer when grown at the lower temperature. Our findings suggest that optimization of titer requires careful consideration of the culture conditions, which should be individualized for the vector producer cell line. PMID- 11044912 TI - Efficient transformation of primary human amniocytes by E1 functions of Ad5: generation of new cell lines for adenoviral vector production. AB - Primary human cells are relatively refractory to transformation by adenoviral E1 functions. For almost two decades, human embryonic kidney (HEK)-derived 293 cells have been the only E1-complementing cell line suitable for production of E1 deleted adenoviral vectors. More recently, new vector production cell lines have been derived from human embryonic retina (HER) cells, a cell type that is difficult to obtain. We were surprised to find that readily available primary human amniocytes are efficiently transformed by adenoviral E1 functions. We selected cell lines that allow high-titer production of recombinant adenoviral vectors. The generation of replication-competent adenovirus (RCA) during production, caused by homologous recombination between vector and cellular DNA, was excluded by designing the transforming plasmid to lack sequence overlap with current adenoviral vectors. In addition, we generated an infectious plasmid that can be used for convenient generation of first-generation adenoviral vectors in Escherichia coli and that matches the E1 complementation in the new production cell lines. PMID- 11044913 TI - Treatment of a lysosomal storage disease, mucopolysaccharidosis VII, with microencapsulated recombinant cells. AB - Most lysosomal enzyme deficiencies are catastrophic illnesses with no generally available treatments. We have used the beta-glucuronidase-deficient mouse model of mucopolysaccharidosis type VII (MPS VII) to develop an alternative approach to therapy. A "universal" cell line engineered to secrete the missing enzyme is implanted in all recipients requiring the same enzyme replacement. The cells, although nonautologous, are rendered immunologically tolerant by encapsulation in microcapsules that provide protection from immune mediators. Using this strategy, we injected beta-glucuronidase-secreting fibroblasts enclosed in alginate microcapsules into mutant MPS VII mice. After 24 hr, beta-glucuronidase activity was detected in the plasma, reaching 66% of physiological levels by 2 weeks postimplantation. Significant beta-glucuronidase activity was detected in liver and spleen for the duration of the 8-week experiment. Concomitantly, the intralysosomal accumulation of undegraded glycosaminoglycans was dramatically reduced in liver and spleen tissue sections and urinary glycosaminoglycan content was reduced to normal levels. Elevated secondary lysosomal enzymes beta hexosaminidase and alpha-galactosidase were also reduced. However, implanted mutant MPS VII mice developed antibodies against the murine beta-glucuronidase, demonstrating a potential obstacle in patients with a null mutation who react against the replaced enzyme as a foreign antigen. The antibody response was transiently circumvented with a single treatment of purified anti-CD4 antibody coadministered with the microcapsules. This resulted in increased levels and duration of beta-glucuronidase delivery. Similarly, treated heterozygous mice maintained elevated levels of beta-glucuronidase and did not develop antibodies. This novel cell-based therapy demonstrates a potentially cost-effective and nonviral treatment applicable to all lysosomal storage diseases. PMID- 11044914 TI - Stimulation of mouse bone marrow cells with kit ligand, FLT3 ligand, and thrombopoietin leads to efficient retrovirus-mediated gene transfer to stem cells, whereas interleukin 3 and interleukin 11 reduce transduction of short- and long-term repopulating cells. AB - The effects of cytokine stimulation during retroviral transduction on in vivo reconstitution of mouse hematopoietic stem cells was tested in a murine competitive repopulation assay with alpha-thalassemia as a marker to distinguish donor and recipient red blood cells (RBCs) and the enhanced green fluorescent protein (EGFP) as a marker for gene transfer. After transplantation, EGFP was detected in up to 90% of circulating RBCs, platelets, and leukocytes, and in primitive progenitors in bone marrow (BM), spleen, and thymus of individual transplanted mice for observation periods of more than 6 months. Large quantitative differences in reconstitution were observed after transplantation with graded numbers (1000-30, 000) of EGFP(+) cells preconditioned with various combinations of Kit ligand (KL), FLT-3 ligand (FL), thrombopoietin (TPO), interleukin 3 (IL-3), and IL-11. Relative to nonmanipulated BM cells, repopulation of EGFP(+) cells was maintained by KL/FL/TPO stimulation, but approximately 30-fold reduced after KL/FL/TPO/IL-3, or KL/FL/IL-3/IL-11. These differences were not caused by changes in the ability of immature hematopoietic cells to home to the BM, which was only moderately reduced. In conclusion, these quantitative transplantation studies of mice demonstrate the importance of optimal ex vivo cytokine stimulation for gene transfer to stem cells with retention of their in vivo hematopoietic potential, and also emphasize that overall in vitro transduction frequency does not predict gene transfer to repopulating stem cells. PMID- 11044916 TI - Department of Health and Human Services National Institutes of Health Recombinant DNA Advisory Committee. Minutes of meeting March 8-10, 2000. PMID- 11044915 TI - Tissue repair with a therapeutic transcription factor. AB - The healing of tissue involves a wide range of molecular, cellular, and physiological events that are coordinated in a temporally specific manner. The cellular transcription factor early growth response factor 1 (Egr-1) is expressed minutes after acute injury and serves to stimulate the production of a class of growth factors whose role is to promote tissue repair. We have studied the effects of Egr-1 expression at the site of dermal wounding in rodents. We find that Egr-1 promotes angiogenesis in vitro and in vivo, increases collagen production, and accelerates wound closure. These results show that Egr-1 gene therapy accelerates the normal healing process and raises the potential use of this therapeutic transcription factor for any aspect of tissue repair. PMID- 11044917 TI - Regulatory issues: future meeting dates and protocol submission deadlines for public recombinant DNA advisory committee (RAC) review PMID- 11044918 TI - Corrigendum PMID- 11044919 TI - Ultrasound study of the pleura. AB - This review describes the sonographic morphology of the most important pleural diseases and defines the role of sonography compared with chest radiography and CT in diagnostic work-up of pleural pathology. PMID- 11044920 TI - Imaging of blunt chest trauma. AB - In western European countries most blunt chest traumas are associated with motor vehicle and sport-related accidents. In Switzerland, 39 of 10,000 inhabitants were involved and severely injured in road accidents in 1998. Fifty two percent of them suffered from blunt chest trauma. According to the Swiss Federal Office of Statistics, traumas represented in men the fourth major cause of death (4%) after cardiovascular disease (38%), cancer (28%), and respiratory disease (7%) in 1998. The outcome of chest trauma patients is determined mainly by the severity of the lesions, the prompt appropriate treatment delivered on the scene of the accident, the time needed to transport the patient to a trauma center, and the immediate recognition of the lesions by a trained emergency team. Other determining factors include age as well as coexisting cardiac, pulmonary, and renal diseases. Our purpose was to review the wide spectrum of pathologies related to blunt chest trauma involving the chest wall, pleura, lungs, trachea and bronchi, aorta, aortic arch vessels, and diaphragm. A particular focus on the diagnostic impact of CT is demonstrated. PMID- 11044921 TI - Focal airtrapping at expiratory high-resolution CT: comparison with pulmonary function tests. AB - This study was undertaken to determine prevalence, extent, and severity of focal airtrapping at expiratory high-resolution CT, and to compare focal airtrapping with age, gender, pulmonary function tests, and blood gas analysis. Two-hundred seventeen patients with and without pulmonary disease underwent paired inspiratory/expiratory high-resolution CT. Six scan pairs with corresponding scan levels were visually assessed for focal--not diffuse--airtrapping using a four point scale. Pulmonary function tests and blood gas analysis were available for correlation in all patients (mean interval 5 days). Focal airtrapping with lower lung predominance was observed in 80% of patients. Twenty-six of 26 patients with restrictive lung function impairment exhibited focal airtrapping (mean score 2.4), whereas only 72 of 98 (74%) patients with obstruction did (mean score 1.5; p < 0.05). Fifty-eight of 70 (83%) patients with normal lung function (mean score 1.8) and 19 of 23 (83%) patients with mixed impairment (mean score 1.8) had focal airtrapping. Focal airtrapping showed negative correlations with static lung volumes (-0.27 to -0.37; p < 0.001) in all patients and moderate positive correlations with dynamic parameters (0.3-0.4; p < 0.001) in patients with obstruction. No significant correlations were found with age, gender, and blood gas analysis. Visual assessment of focal--not diffuse--airtrapping at expiratory high-resolution CT does not correlate with physiological evidence of obstruction as derived from pulmonary function tests since the perception of focal airtrapping requires an adequate expiratory increase in lung density. PMID- 11044922 TI - Pseudomembranous aspergillus bronchitis in a double-lung transplanted patient: unusual radiographic and CT features. AB - Pseudomembranous aspergillus bronchitis is considered as an early form of invasive pulmonary aspergillosis, a well-known airway infection in immunocompromised patients. Radiologic features concerning invasive aspergillosis of the airways have been reported. However, we describe here an unusual feature of invasive aspergillus bronchitis, never reported to date, observed in a double lung transplanted patient. Chest radiograph and CT revealed significant peribronchial thickening without any parenchymal involvement. PMID- 11044923 TI - Electrical impedance measurement of the breast: effect of hormonal changes associated with the menstrual cycle. AB - The purpose of this study was to evaluate the influence of hormonal factors on electrical impedance measurement with a new device TS2000, which is a new method in diagnosis of breast disease. Twenty-one healthy pre-menopausal women volunteers (aged 24-39 years) were examined with the TS2000 once/week for two menstrual cycles. On average, at least one spot was present in 47% of images of women not taking oral contraceptives and in 44% of women taking oral contraceptives (OC). The number of spots varied over the menstrual cycle and had a maximum in week 3 and week 5. We found that after 1 week only 15% of spots were present and no spots persisted for three consecutive weeks. These data, if further supported by observations on other populations of women, show that false positive results are common in pre-menopausal women; however, these false positive spots do not persist for long periods of time. This information may provide a basis for discrimination between true-positive and false-positive spots on the TS2000 image, since the latter would be expected to disappear on short term follow up. PMID- 11044925 TI - Ultrasound variables and their prognostic value in a population of 1103 patients with 272 breast cancers. AB - In a prospective two-centre study targeted US was performed as an adjunct to mammography in a population of 1103 patients with 272 breast cancers, 517 benign lesions and no abnormalities in 314 patients. The purpose of the study was to analyse the distribution of the different US variables among the breast lesions and to determine the prognostic value of these variables with respect to the diagnosis of malignancy. The following variables were analysed: border; contour; orientation; structure; echogenicity; sound transmission; and size. These variables were correlated with the definitive diagnosis and univariate analysis was performed. A statistically significant association with breast cancer (p < 0.001) was present for irregular border; ill-defined contour; indeterminate or vertical orientation; homogeneous, complex or heterogeneous structure; hypoechogenicity; and unchanged or decreased sound transmission. Multivariate analysis showed a high independent prognostic value for malignancy for irregular border, followed by ill-defined contour and unchanged or decreased sound transmission. However, their discriminative power was not absolute: in 38 cancers none of the malignant US variables were present, and in 11 benign lesions all variables were present. There were hardly any benign-looking lesions that proved to be malignant. PMID- 11044924 TI - Electrical impedance scanning for classifying suspicious breast lesions: first results. AB - It has long been established that cancer cells exhibit altered local dielectric properties compared with normal cells. Consequently, different electrical conductivity and capacitance are measurable in malignant vs normal tissues. In this study we evaluated the reliability of electrical impedance scanning (EIS), a new technology, for the classification of suspicious lesions: differentiating benign from malignant, and as a primary means of detection of breast cancer. Fifty-two women with 58 sonographically and/or mammographically suspicious findings were examined using electrical impedance scanning. Two different examination modes of TransScan TS2000 (Siemens, Erlangen, Germany), the standard resolution mode for a routine overview examination, and the targeted high resolution mode for a local examination of the suspicious lesion were used. All patients were additionally imaged by MR mammography (MRM) and underwent core biopsy and/or surgical treatment after the EIS examination. With respect to the histopathological findings (29 malignant and 29 benign lesions) 27 of 29 (93.1%) malignant lesions were correctly identified using the high-resolution mode of EIS, whereas 19 of 29 (65.5%) benign lesions were correctly identified as benign (10 of 29 benign lesions showed as false-positive findings). Negative and positive predictive values of 90.5 and 73.0% were observed, respectively. Using the standard-resolution mode 22 of 29 malignancies were correctly detected (sensitivity 75.9%), whereas 22 of 29 were correctly identified as benign (specificity 72.4%). Electrical impedance scanning appears to be a promising new technology providing a relatively high sensitivity for the verification of suspicious mammographic and/or sonographic lesions especially using the high resolution mode for local examinations. Artifacts, such as signals from superficial skin lesions, poor contact, and air bubbles, are currently a limitation. PMID- 11044926 TI - Therapeutic effect of transcatheter arterial chemoembolization on hepatocellular carcinoma: evaluation with contrast-enhanced harmonic power Doppler ultrasound. AB - The aim of this study was to investigate the usefulness of contrast-enhanced harmonic power Doppler ultrasound (US) for the detection of residual viable hepatocellular carcinoma (HCC) after treatment with transcatheter arterial chemoembolization (TACE). Forty-seven patients with 68 HCC lesions 1.8-9.5 cm in diameter (mean +/- SD 4.3 +/- 1.7 cm) underwent contrast-enhanced power Doppler US, in the harmonic mode, before and after treatment with TACE. Unenhanced spiral CT and contrast-enhanced dynamic MR imaging were also performed to help establish the outcome of therapy. Before treatment, intratumoral blood flow signals were detected at contrast-enhanced harmonic power Doppler US in 65 (95%) of 68 lesions. After TACE, flow signals were no longer detectable in 22 of these 65 lesions, which showed complete response at spiral CT and dynamic MR imaging. In 38 (88%) of the 43 lesions with partial response, intratumoral flow signals were still identified at contrast-enhanced harmonic power Doppler US. Twenty-eight of these 38 lesions underwent additional treatment with percutaneous ethanol injection (PEI) using contrast-enhanced harmonic power Doppler US guidance. Complete response was seen after PEI in 23 of 28 lesions. Contrast-enhanced harmonic power Doppler US proved useful for assessing the therapeutic effect of TACE on HCC and for guiding additional treatment with PEI in cases of partial response. PMID- 11044927 TI - Value of MR cholangiography in the evaluation of postoperative biliary complications following orthotopic liver transplantation. AB - The aim of this study was to describe the spectrum of abnormal biliary findings as seen with magnetic resonance cholangiography (MRC) in symptomatic patients after orthotopic liver transplantation (OLT). In our study we included 12 consecutive patients post-OLT who presented with clinical and/or biochemical suspicion of biliary complications. In all patients MRC was performed on a 1.0-T whole-body magnet and breathhold half-Fourier acquired single-shot turbo spin echo and rapid acquisition with relaxation enhancement sequences were used. Diagnostic confirmation was obtained with percutaneous transhepatic cholangiography (PTC; n = 3 patients), endoscopic retrograde cholangiography (ERC; n = 3 patients), or clinical follow-up. A vast array of biliary abnormalities were detected in 11 of 12 patients: high-grade, obstructive, anastomotic stricture was the most common unique abnormality. Findings consistent with bile duct necrosis, the second most common abnormality, were accompanied by arterial abnormalities in 2 of 5 patients on subsequent MR- and digital subtraction angiography. Compared with the findings obtained with direct cholangiography (n = 5 patients), MRC was highly accurate for the detection and characterization of postoperative biliary complications. Compared with the final diagnosis, which was based on PTC-ERC findings and/or all available clinical data, MRC imaging alone was able to provide a specific diagnosis in 9 of 12 patients. Magnetic resonance cholangiography is an accurate, time-saving, and non invasive imaging modality in the evaluation of post-OLT patients in whom suspicion of biliary complications exists. Although the precise value of MRA in this patient group requires larger dedicated studies, single session "all-in-one" MR evaluation of both biliary and arterial system in our series proved to be a substantial benefit in obtaining an accurate and complete diagnosis. PMID- 11044928 TI - Role of the intra-arterial calcium stimulation test in the preoperative localization of insulinomas. AB - The aim of this study was determination of the significance of the arterial stimulation test with venous sampling (ASVS) in the preoperative localization of insulinoma. Eleven patients with endogenous hyperinsulinism underwent preoperative transabdominal US, spiral computer tomography (spiral CT), MRI, endoscopic ultrasound (EUS) as well as angiography (DSA) combined with ASVS. The results were compared with intraoperative findings, intraoperative ultrasound (IOUS) and histopathology. There were no complications related to the ASVS test. In 11 patients the tumor could be localized with the various modalities as follows: US 1 of 11 (9%), MRI 3 of 10 (30%), spiral CT 4 of 11 (36%), EUS 5 of 10 (50%), DSA 8 of 11 (73%), and ASVS 10 of 11 (91%). In 2 patients the tumors were intraoperatively neither palpable nor detectable by IOUS, and consequently the intraoperative management was governed by information provided by DSA combined with the ASVS test. Ten patients had solitary benign insulinomas and 1 patient with multiple endocrine neoplasia I had two tumors adjacent to each other in the pancreatic tail. Arterial stimulation test with venous sampling was the most sensitive preoperative test for regionalizing the insulinoma in our set of patients. It can be performed safely in the course of a regular DSA examination and may affect intra-operative management in patients in whom the tumors are not detectable by palpation or IOUS. PMID- 11044929 TI - Power Doppler ultrasound of gallbladder wall vascularization in inflammation: clinical implications. AB - We investigated the role of Power Doppler US in the diagnosis and follow-up of cholecystitis. We reviewed the examinations of 21 surgical patients aged 27-48 years with US findings of cholecystitis. We performed B-mode and then Power Doppler US. Wall thickness and US structure, the presence/absence of stones, and US Murphy's sign were assessed at B-mode US, whereas only the presence/absence of wall vascularization was studied with Power Doppler. B-mode and Power Doppler changes post treatment were also investigated. Ultrasound showed wall thickening in all patients. In addition, positive Murphy's sign and/or gallbladder stones were seen in 6 patients each at B-mode US and wall vascularization in 7 patients with Power Doppler. Acute cholecystitis was diagnosed in these patients. The other 14 patients presenting wall thickening but no vascularization and negative US Murphy's sign were diagnosed as having chronic cholecystitis; 10 of them had gallbladder stones. Two of seven acute cholecystitis patients were operated on in the acute stage for the onset of complications and histologic findings confirmed the US diagnosis. As for the remaining patients, histology diagnosed chronic cholecystitis in 17, whereas wall thickening was not inflammatory in 2 cases. All the cases with early wall vascularization were eventually diagnosed as cholecystitis. Power Doppler US permits confirmation of the diagnosis of acute cholecystitis and distinguishing of chronic disease, which helps in planning of surgery. PMID- 11044930 TI - Computed tomography after lymphangiography in the diagnosis of intestinal lymphangiectasia with protein-losing enteropathy in Noonan's syndrome. AB - Noonan's syndrome is a rare congenital disorder that may be associated with abnormalities in the lymphatic drainage. In this case of a 21-year-old man CT after bipedal lymphangiography confirmed the diagnosis of intestinal lymphangiectasy causing protein-losing enteropathy in Noonan's syndrome by showing contrast-enhanced abnormal lymphatic vessels in the mesentery and the intestinal wall. Because of the benefit of diet in case of intestinal involvement, we recommend a thorough documentation of the lymphatic drainage with lymphangiography followed by CT, if clinical signs of lymphatic dysplasia, such as pleural effusions, lymphedema, or hypoproteinemia are present. PMID- 11044931 TI - Value of fluid-attenuated inversion recovery sequences in early MRI of the brain in neonates with a perinatal hypoxic-ischemic encephalopathy. AB - The aim of our study was to assess the usefulness of fluid-attenuated inversion recovery (FLAIR) sequences in comparison with conventional spin-echo and inversion MR imaging in neonates for evaluation of myelination and for detection of hypoxic-ischemic brain injury. We reviewed early MR scans of 18 neonates with suspected hypoxic-ischemic brain damage. Myelination could be evaluated with confidence using conventional MR imaging in all but 2 infants; however, the presence of myelin was very difficult to assess on FLAIR images. Overall, 53 lesions or groups of lesions were identified. The FLAIR technique was more sensitive in 11 of the lesions; especially (pre)cystic lesions could be identified much better and more cysts were found. Conventional MR imaging failed to identify 2 of the lesions and was more sensitive in 14 of the lesions; especially punctate hemorrhages and lesions in basal ganglia or thalami could be better determined. The FLAIR technique missed 3 of these lesions. In the remaining 28 lesions conventional MR and FLAIR images were equally diagnostic. The FLAIR technique and conventional MR imaging are complementary in detecting early sequelae of hypoxic-ischemic brain injury in neonates. The FLAIR technique is not suitable for assessing myelination of the neonatal brain; therefore, FLAIR cannot replace conventional MR imaging. PMID- 11044932 TI - Chronic nontraumatic spinal epidural hematoma of the lumbar spine: MRI diagnosis. AB - An uncommon case of chronic nontraumatic spinal epidural hematoma of the lumbar spine in a 75-year-old woman is reported. The patient presented with a 7-month history of low back pain and bilateral sciatica. Magnetic resonance imaging enabled a correct preoperative diagnosis revealing a nodular, well-circumscribed epidural mass with peripheral enhancement and signal intensities consistent with chronic hematoma, which extended from L2 to L3. Laminectomy of L2-L3 was performed and the hematoma was totally resected. Histological examination of the surgical specimen demonstrated a chronic encapsulated hematoma. No evidence of vascular malformation was found. The patient recovered fully after surgical treatment. PMID- 11044933 TI - Intracranial dural arteriovenous fistula with spinal medullary venous drainage. AB - We report on a 46-year-old patient in whom an intracranial dural arteriovenous (AV) fistula, supplied by a branch of the ascending pharyngeal artery, drained into spinal veins and produced rapidly progressive symptoms of myelopathy and brainstem dysfunction including respiratory insufficiency. Magnetic resonance imaging studies demonstrated brainstem oedema and dilated veins of the brainstem and spinal cord. Endovascular embolization of the fistula led to good neurological recovery, although the patient had been paraplegic for 24 h prior to embolization. This case demonstrates the MRI characteristics of an intracranial dural AV fistula with spinal drainage and illustrates the importance of early diagnosis and treatment. Even paraplegia may be reversible, if angiography is performed and the fistula treated before ischaemic and gliotic changes become irreversible. PMID- 11044934 TI - Congenital infiltrating lipomatosis of the face with cerebral abnormalities. AB - The aim of this study was to describe a possible variant of encephalo craniocutaneous lipomatosis syndrome. Three cases of congenital infiltrating lipomatosis of the face, associated with cutaneous, subcutaneous, and cerebral abnormalities, are presented. This neurocutaneous syndrome appears very similar to encephalo-craniocutaneous lipomatosis syndrome but lacks the typical eye lesions. PMID- 11044935 TI - Magnetic resonance imaging in the assessment of urologic disease: an all-in-one approach. AB - The aim of this study was to evaluate an "all-in-one" MR procedure to examine the kidneys, the renal vascular supply and renal perfusion, and the urinary tract. In 64 patients (58 with urologic disease and 6 healthy volunteers), MR was performed including: (a) T1- and T2-weighted imaging; (b) 3D contrast-enhanced MR angiography (MRA), including the renal arteries, renal veins, as well as renal perfusion; and (c) 3D contrast-enhanced MR urography (MRU) in the coronal and sagittal plane. For the latter, low- and high-resolution images were compared. Prior to gadolinium injection, 0.1 mg/kg body weight of furosemide was administered intravenously. The results were compared with correlative imaging modalities (ultrasonography, intravenous urography, CT), ureterorenoscopy and/or surgical-pathologic findings. Visualization of the renal parenchyma, the vascular supply, and the collecting system was adequate in all cases, both in nondilated and in dilated systems and irrespective of the renal function. One infiltrating urothelial cancer was missed; there was one false-positive urothelial malignancy. Different MR techniques can be combined to establish an all-in-one imaging modality in the assessment of diseases which affect the kidneys and urinary tracts. Continuous refinement of the applied MR techniques and further improvements in spatial resolution is needed to expand the actual imaging possibilities and to create new tracts and challenges in the MR evaluation of urologic disease. PMID- 11044936 TI - Cost analysis of different protocols for imaging a patient with acute flank pain. AB - The aim of this study was to analyse the costs of different diagnostic approaches to patients with acute flank pain. Four different diagnostic approaches were considered: (a) spiral CT without contrast medium (CM); (b) plain film, ultrasonography (US) and intravenous urography (IVU)--the latter procedure is used in our department in cases still unsolved following the former investigations (28% in our experience); (c) plain film, US and spiral CT without CM (as an alternative to IVU in 28% of cases); and (d) IVU. The cost of each procedure in a university hospital was calculated, following analysis of the differential costs of each investigation (equipment, depreciation and maintenance costs, related materials and services, radiologists, radiographers, nurses) and their common costs (auxiliary personnel and indirect internal costs). Finally, we calculated the full cost of each procedure and applied it to the different diagnostic approaches. The full cost of each approach was: (a) spiral CT without CM = 74 Euro; (b) plain film, US and IVU (28%) = 66.89 Euro; (c) plain film, US and spiral CT without CM (28%) = 64.93 Euro; (d) IVU = 80.90 Euro. Intravenous urography alone or in unsolved cases is not to be considered because it provides higher costs and worse diagnostic results, whereas X-ray dose to patient is almost equal between IVU and spiral CT. Spiral CT integrated to plain film and US in unsolved cases could be preferred because of lower cost and dose to patient, though reaching a diagnostic conclusion may take longer than an immediate spiral CT. PMID- 11044937 TI - Non-neoplastic intratesticular lesions mimicking tumour on ultrasound. AB - It has recently been emphasized that the incidence of benign testicular lesions is much higher than previously suspected. Sonography is reported to have a high sensitivity but poor specificity in the evaluation of intratesticular abnormalities. This report reviews the common and unusual lesions that can mimic testicular tumour on ultrasound, and discusses the clinical and sonographic features that may help to narrow the differential diagnosis and guide management. PMID- 11044938 TI - Fibrous pseudotumor of the epididymis: imaging and pathologic correlation. AB - We report the case of a 25-year-old man who presented a fibrous pseudotumor of the epididymis, a rare focal location of nodular and diffuse fibrous proliferation. We provide the ultrasonographic and MRI findings with pathologic correlation. PMID- 11044939 TI - Unusual retrovesical masses in men. AB - Retrovesical masses in men not related to prostatic carcinoma or hyperplasia is an uncommon pathology. Rare masses or unusual manifestations of those common diseases are a diagnostic dilemma. We review our experience in three unusual retrovesical masses in men: carcinosarcoma filling a giant bladder diverticulum; cystic prostatic carcinoma; and acquired cystic dilatation of the seminal vesicle associated with a prostatic carcinoma that obstructed and invaded the vesicle. We report the imaging findings and review the literature. In our experience, the imaging findings are usually not specific for doing a precise diagnosis and biopsy procedures are necessary. PMID- 11044940 TI - Doppler sonographic renal resistance index in healthy children. AB - The objective of the study was to determine the values of Doppler resistance indices (RIs) in intrarenal arteries in healthy children. Color duplex Doppler sonography of intrarenal arteries was performed in 163 children and 58 adult patients, with the absence of clinical or laboratory pathologic changes of the urinary tract. All patients were classified following results of exploratory data analysis into three age groups: (a) 51 children between 2 and < 6 years of age; (b) 112 children 6-16 years of age; and (c) adults. The RIs were compared between different age groups. The mean RI +/- 1 SD values in group 1 were 0.705 +/- 0.018 (range 0.67-0.75), in group 2 0.605 +/- 0.029 (range 0.55-0.65), and in the adult group 0.604 +/- 0.035 (range 0.54-0.68). In the group of the youngest children (group 1), the RIs were significantly higher (p < 0.01) in comparison with the other group of children and the adult group. No statistically significant difference was observed between the group of children 6-16 years of age and the adult group. RI < 0.70 was noted in all children above 6 years of age; however, in younger children, RI > or = 0.70 was observed in 72.5% (37 of 51) of children. Resistance index in early childhood was considerably higher as compared with older children and the adult population: Apparently the RI value drops to adult level at approximately 6 years of age. The utilization of a threshold RI value of 0.70 for the increased renal vascular resistance in adults can also be applied to children over 6 years of age. PMID- 11044941 TI - Thoracoabdominal peripheral primitive neuroectodermal tumors in childhood: radiological features. AB - Peripheral primitive neuroectodermal tumors (PNET) are extremely uncommon, malignant neoplasms affecting mostly children and young adults. We retrospectively reviewed the clinical data and radiological studies of four such cases. All cases were pathologically proven. Plain films, US, and CT scans were used. The youngest child had a huge pelvic tumor and two adolescents each had a chest wall (Askin) tumor. The fourth patient had a most unusual location of the PNET in the anterior mediastinum. The CT findings are emphasized. We emphasize that the markedly abnormal CT findings are not specific for PNET. PMID- 11044942 TI - Cystic dysplasia of the testis: a rare cause of painless hemiscrotal enlargement in childhood. AB - Cystic dysplasia of the testis is a rare, benign cause of painless hemiscrotal enlargement in children. We present two cases of cystic dysplasia of the testis presenting with scrotal enlargement. The diagnosis was based on its specific sonographic findings and on the coexistence of a multicystic dysplastic kidney in one case. PMID- 11044943 TI - MRI of the plantar structures of the foot after falanga torture. AB - Falanga is an ancient form of punishment or torture but is still commonly reported by our refugees. The late result of caning the heel and ball of the foot is a chronic painful condition with few clinical signs. The aim of the present study was to assess, by MRI, possible morphologic characteristics of the heel and ball of the foot, related to falanga and pain in correlation to clinical findings. Magnetic resonance imaging of the foot was obtained in 12 victims exposed to falanga torture and 9 healthy volunteers. Sagittal T1-weighted spin echo images (TR 616-840 ms, TE 20 ms), T2-weighted spin-echo images (TR 1900 ms, TE 90 ms), and short tau inversion recovery (STIR) images (TR 1200 ms, TE 15 ms, TI 100 ms) were performed. The central portion of the plantar aponeurosis was generally significantly thicker in victims exposed to falanga torture as compared with that of controls (P < 0.05). In all except one of the victims, MRI demonstrated two layers of the thickened plantar aponeurosis: a deeper portion with normal homogeneous low signal intensity (SI) appearance, and a superficial layer with characteristic areas of mixed SI on both T1- and T2-weighted images. There were no signs of chronic muscular compartment syndromes, and the thickness of the plantar pad did not differ between the two groups. Magnetic resonance imaging may demonstrate morphologic characteristics of the plantar aponeurosis which may confirm falanga torture. Further imaging with more specific sequences is warranted to demonstrate the supposed injuries in the compartmental fat tissue chambers and the vascularity of the ball pad of the foot. PMID- 11044944 TI - Extraosseous manifestation of Gaucher's disease type I: MR and histological appearance. AB - Gaucher's disease type I is the most prevalent lysosomal storage disorder caused by an autosomal-recessive inherited deficiency of glucocerebrosidase activity with secondary accumulation of glucocerebrosides within the lysosomes of macrophages. The storage disorder produces a multisystem disease characterized by progressive visceral enlargement and gradual replacement of bone marrow with lipid-laden macrophages. Skeletal disease is a major source of disability in Gaucher's disease. Extraosseous extension of Gaucher cells is an extremely rare manifestation of skeletal Gaucher's disease. This is a report on the MRI and histopathological findings of an extraosseous Gaucher-cell extension into the midface in a patient with Gaucher's disease. PMID- 11044945 TI - Pseudosarcoma in Paget's disease. AB - The authors describe the case of a male patient who had had Paget's disease since 1973 which led to the diffuse swelling of the proximal femur in 1993. The elevated alkaline phosphatase level and the destructive lytic lesions on the X ray of the femur raised the possibility of malignancy. The bone scan showed increased accumulation in the proximal part of the left femur. On the MR the low T1 and the elevated T2 signal intensity suggested malignity despite that in some regions signals typical of adipose tissue could also be seen, which is usually interpreted as a sign of benignity. In the histological specimens no signs of malignant tumour cells were identified. The patient received bisphosphonate treatment which decreased his pain and swelling and also the alkaline phosphatase level. The patient died because of a rapidly progressing retroperitoneal fibrosarcoma and lung metastasis in 1996. The autopsy did not prove any malignant transformation of Paget's disease of bone in the proximal femur. PMID- 11044946 TI - Masses of the tongue and floor of the mouth: findings on magnetic resonance imaging. AB - The purpose of this study was to determine the value of MR imaging for the demonstration of masses in the tongue and floor of the mouth. Nine patients were prospectively examined with MR imaging after physical examination. Imaging protocol included T2 and contrast-enhanced and non-contrast-enhanced T1-weighted turbo spin-echo sequences, and the findings were compared with surgical and histopathological results. Histopathological examination revealed four squamous cell carcinomas, one adenoid cystic carcinoma, two tongue abscesses, and one chronic inflammatory change. The other case was diagnosed as hemangioma depending on clinical and imaging findings alone. In cases with squamous cell carcinoma, staging was done on the basis of MR imaging findings, and was found to be T4 in two cases, T3 in one case, and T2 in another. The primary role of MR imaging of the tongue and oropharynx is not to make a tissue diagnosis. Multiple deep biopsies are mandatory for the differentiation of other inflammatory and neoplastic lesions. Magnetic resonance imaging produces coronal and sagittal image planes to assess the volume and spread of the lesion and helps the surgeon determine the direction in which the biopsy should be performed. PMID- 11044947 TI - MRI-visible pericochlear lesions in osteogenesis imperfecta type I. AB - Osteogenesis imperfecta (OI) is an inherited generalized disorder of type-I collagen synthesis often associated with hearing loss. We present a case of OI type I in which hearing loss led to examination of the temporal bone with MRI. In the osseous otic capsule MRI demonstrated pericochlear lesions with soft tissue signal intensity and contrast enhancement. Changes similar to otosclerosis have been described in the temporal bone of OI patients when applying CT, but reports on MRI findings do not yet exist. PMID- 11044949 TI - An unusual case of obstruction of the terminal ileum. PMID- 11044948 TI - Chondromyxoid fibroma of the temporal bone: CT and MRI findings. AB - We present the case of a 44-year-old woman with chondromyxoid fibroma of temporal bone origin. Since this is the least common bone tumor of cartilaginous origin, it is highly unusual to find this tumor in the skull. In fact, the literature describes 18 cases of this form of neoplasia arising in the skull, only 4 of these having originated in the temporal bone. To date, the radiological features of these tumors, and especially features detected using the latest imaging modalities, have not been described in detail. This report is unique in that it is the first to present a case of chondromyxoid fibroma of the temporal bone accompanied by detailed CT and MRI findings. PMID- 11044950 TI - Mann's syndrome: inferior vena cava thrombosis due to aortic aneurysm. PMID- 11044951 TI - Intracranial and extracranial MR angiography in occipital horn syndrome. PMID- 11044952 TI - Pulmonary metastases from pancreatic adenocarcinoma mimicking bronchoalveolar carcinoma. PMID- 11044953 TI - Midgut volvulus in an adolescent patient. PMID- 11044954 TI - Pediatric liver neoplasms. PMID- 11044955 TI - Quiz case of the month. PMID- 11044956 TI - The effectiveness of domiciliary health visiting: a systematic review of international studies and a selective review of the British literature. PMID- 11044957 TI - Routine referral for radiography of patients presenting with low back pain: is patients' outcome influenced by GPs' referral for plain radiography? PMID- 11044958 TI - An interview with Professor Jon Nicholl. PMID- 11044959 TI - National Electronic Library for Health--emergency care. PMID- 11044960 TI - Got Health? The Hawaii Partnership for Standards-Based School Health Education. AB - The American Cancer Society (ACS), Hawaii Pacific, Inc., initiated the Hawaii Partnership for Standards-Based School Health Education in July 1999. The goal of the partnership is to implement standards-based school health education to promote child and adolescent health in Hawaii. The partnership includes representatives from the Hawaii Department of Education, the Hawaii Board of Education, the Hawaii Department of Health, the University of Hawaii College of Education, Meadow Gold Dairies, the Hawaiian Electric Companies, the Hawaii Medical Services Association, and local divisions of the American Cancer Society. During its first year, the partnership collaborated to sponsor 15 teacher workshops on the new Hawaii Health Content Standards. In addition, Meadow Gold Dairies initiated a Got Health? milk carton side-panel promotional campaign to publicize the standards. This article describes development of the Hawaii Health Content Standards, history of the partnership, activities to date, concurrent supporting efforts, future plans, and lessons learned during the first year. PMID- 11044962 TI - Code of Ethics for the Health Education profession. PMID- 11044961 TI - Development of a Unified Code of Ethics for the Health Education Profession: a report of the National Task Force on Ethics in Health Education. PMID- 11044963 TI - An approach to teaching for critical thinking in health education. AB - Teaching for critical thinking represents a vital and emerging priority in school health education. A variety of conceptions of critical thinking and approaches to teaching for critical thinking exist in the literature. This paper explores the relevance of Richard Paul's concept of critical thinking to health education. Paul's work on critical thinking has been widely disseminated and features three inter-related components called the elements of reasoning, intellectual standards, and intellectual traits. Each component appears highly relevant to health education. Paul's approach is also based on natural rather than technical language which improves its utility in learning. Unanswered questions about the approach concern the degree to which the thinking skills and traits can be transferred to health instruction and the lack of evaluation research demonstrating its efficacy. Further inquiry into the applicability and efficacy of the approach is needed. PMID- 11044965 TI - Politics and the success of school-based health centers. AB - School-based health centers (SBHCs) provide access to health services by bringing providers to children (and sometimes parents) and furnishing low cost services in an atmosphere of trust. While the number of SBHCs has continued to grow and some clinics have continued to expand their services, others have barely survived and some have even closed. This study investigated factors, particularly political forces, that affected the success of SBHCs. Using a national survey of clinic directors, this study assessed clinic success in terms both of longevity and service delivery. Findings indicate the factors most consistently and significantly associated with success include not only measures of "need" (school size and percent African-American enrollment or population) but of "politics" (citizen political ideology and Southern conservatism). Thus, politics matters more than previous studies suggested. PMID- 11044964 TI - The association between severity of sanction imposed for violation of tobacco policy and high school dropout rates. AB - This investigation explored the association between severity of sanctions imposed on students resulting from tobacco policy violation and the event dropout rate in South Carolina public high schools. The study employed a cross-sectional design (n = 132). Surveys were mailed to school principals to assess tobacco policy and sanctions for violation. Severe sanctions were categorized as those resulting in the student being denied onsite instruction, such as out-of-school suspension or expulsion. General linear regression models adjusting for SES, ethnicity, and rural/urban status, tested for an association between event dropout rate and severity of sanction imposed. The mean dropout rate in 1998 for high schools in South Carolina was 2.58% (+1.74). Suspension at first violation and expulsion were associated with lower dropout rates. Suspension at second violation was not associated with dropout behavior while suspension at third violation was associated with higher dropout rates. Results from the study provide preliminary evidence that severe sanctions imposed for violation of tobacco policy may help reduce high school dropout rates. PMID- 11044966 TI - Rural parents' communication with their teen-agers about sexual issues. AB - This survey assessed rural parents' (n = 374) perceptions of the characteristics, content, and comfort level of discussions about sexual issues with their teens. Almost all parents (94%) reported they had talked with their teens about sex. Two thirds (65%) reported being comfortable talking with their teens about sexual issues. From a list of 17 potential topical areas in sexual communication, parents were most likely to discuss with their teens the responsibilities of being a parent (46%), sexually transmitted diseases (40%), dating behavior (37%), and not having sex until marriage (36%). Most parents (80%) believed that the majority of sexuality education should be provided by the family and supplemented by outside organizations, preferably schools. Almost all parents (92%) believed sexuality education should include information on birth control methods including condoms. Almost two of three parents (64%) believed schools should begin teaching sexuality education before students reach seventh grade. Parents (52%) claimed they could best be helped in communicating with their teens by receiving a regular newsletter regarding teen sexual issues. PMID- 11044967 TI - Analyzing health-related election issues. PMID- 11044968 TI - Evaluating student services provided by school-based health centers: applying the Shuler Nurse Practitioner Practice model. PMID- 11044969 TI - Recommendations for the outpatient surveillance of renal transplant recipients. American Society of Transplantation. AB - Many complications after renal transplantation can be prevented if they are detected early. Guidelines have been developed for the prevention of diseases in the general population, but there are no comprehensive guidelines for the prevention of diseases and complications after renal transplantation. Therefore, the Clinical Practice Guidelines Committee of the American Society of Transplantation developed these guidelines to help physicians and other health care workers provide optimal care for renal transplant recipients. The guidelines are also intended to indirectly help patients receive the access to care that they need to ensure long-term allograft survival, by attempting to systematically define what that care encompasses. The guidelines are applicable to all adult and pediatric renal transplant recipients, and they cover the outpatient screening for and prevention of diseases and complications that commonly occur after renal transplantation. They do not cover the diagnosis and treatment of diseases and complications after they become manifest, and they do not cover the pretransplant evaluation of renal transplant candidates. The guidelines are comprehensive, but they do not pretend to cover every aspect of care. As much as possible, the guidelines are evidence-based, and each recommendation has been given a subjective grade to indicate the strength of evidence that supports the recommendation. It is hoped that these guidelines will provide a framework for additional discussion and research that will improve the care of renal transplant recipients. PMID- 11044970 TI - Epitope tagging: general method for tracking recombinant proteins. AB - Epitope tagging has provided a useful experimental strategy with widespread applicability. The ample variety of epitope tag systems that have been put to use to date provide a collection of attributes relevant to virtually any experimental system. As a consequence, epitope tagging will continue to be a valuable tool for molecular biologists long into the future. PMID- 11044971 TI - Alkaline phosphatase fusions of ligands or receptors as in situ probes for staining of cells, tissues, and embryos. PMID- 11044972 TI - Chimeric molecules employing horseradish peroxidase as reporter enzyme for protein localization in the electron microscope. PMID- 11044973 TI - Biochemical analyses of trafficking with horseradish peroxidase-tagged chimeras. PMID- 11044974 TI - Visualizing protein dynamics in yeast with green fluorescent protein. PMID- 11044975 TI - Kinetic analysis of intracellular trafficking in single living cells with vesicular stomatitis virus protein G-green fluorescent protein hybrids. PMID- 11044976 TI - Dual color detection of cyan and yellow derivatives of green fluorescent protein using conventional fluorescence microscopy and 35-mm photography. PMID- 11044977 TI - Invertase fusion proteins for analysis of protein trafficking in yeast. PMID- 11044978 TI - Introduction of Kex2 cleavage sites in fusion proteins for monitoring localization and transport in yeast secretory pathway. PMID- 11044979 TI - Lineage analysis with retroviral vectors. PMID- 11044980 TI - Use of pseudotyped retroviruses in zebrafish as genetic tags. PMID- 11044981 TI - A gene fusion method for assaying interactions of protein transmembrane segments in vivo. PMID- 11044982 TI - Using SUC2-HIS4C reporter domain to study topology of membrane proteins in Saccharomyces cerevisiae. PMID- 11044983 TI - Detecting interactions between membrane proteins in vivo using chimeras. PMID- 11044984 TI - Alkaline phosphatase fusion proteins for molecular characterization and cloning of receptors and their ligands. PMID- 11044985 TI - Surface chimeric receptors as tools in study of lymphocyte activation. AB - In this chapter we have described a powerful technology that has allowed the functional dissection of individual subunits from oligomeric receptors. We have focused primarily on chimeras derived from antigen receptors or their downstream signaling components to illustrate the wide utility of the approach; however, the technology has been applied to numerous multimeric receptors of the immune system including cytokine receptors, Fc receptors, and natural killer (NK) cell inhibitory receptors. Although the significance of the structural complexity of oligomeric receptors is by no means understood, it is certain that valuable benefits must be derived from the integrated function of their subunits. In the case of antigen receptors, the multiplicity of ITAMs likely allows the cell to distinguish subtle variations in ligand affinities with exquisite sensitivity. Clearly, an isolated subunit that is ligated with antibodies cannot confer such complex function. For instance, it cannot reveal the subtle changes in signal transduction that likely occur on stimulation with altered antigenic peptide ligands or during a complex cell-cell interaction. However, before the intricacies of integrated receptor function can be appreciated, the potential or unique functional properties contributed by each individual receptor component must first be understood. Providing a tool to acquire this kind of understanding has been the greatest asset of this technology. Acknowledging its limitations, the use of surface chimeric receptors remains an invaluable approach toward our understanding the complex function of oligomeric receptors. PMID- 11044986 TI - Use of chimeric receptor molecules to dissect signal transduction mechanisms. PMID- 11044987 TI - Fusion protein toxins based on diphtheria toxin: selective targeting of growth factor receptors of eukaryotic cells. PMID- 11044988 TI - Green fluorescent protein-based sensors for detecting signal transduction and monitoring ion channel function. PMID- 11044989 TI - Metabolic labeling of glycoproteins with chemical tags through unnatural sialic acid biosynthesis. PMID- 11044990 TI - Using sorting signals to retain proteins in endoplasmic reticulum. PMID- 11044991 TI - Directing proteins to nucleus by fusion to nuclear localization signal tags. PMID- 11044992 TI - Identification, analysis, and use of nuclear export signals in Saccharomyces cerevisiae. PMID- 11044993 TI - Directing proteins to mitochondria by fusion to mitochondrial targeting signals. PMID- 11044994 TI - Targeting proteins to plasma membrane and membrane microdomains by N-terminal myristoylation and palmitoylation. PMID- 11044995 TI - Analysis of function and regulation of proteins that mediate signal transduction by use of lipid-modified plasma membrane-targeting sequences. AB - It is now established that the function of many signaling molecules is controlled, in part, by regulation of subcellular localization. For example, the dynamic recruitment of normally cytosolic proteins to the plasma membrane, by activated Ras or activated receptor tyrosine kinases, facilitates their interaction with other membrane-associated components that participate in their full activation (e.g., Raf-1). Therefore, the creation of chimeric proteins that contain lipid-modified signaling sequences that direct membrane localization allows the generation of constitutively activated variants of such proteins. The amino-terminal myristoylation signal sequence of Src family proteins and the carboxy-terminal prenylation signal sequence of Ras proteins have been widely used to achieve this goal. Such membrane-targeted variants have proved to be valuable reagents in the study of the biochemical and biological properties of many signaling molecules. PMID- 11044996 TI - Glycerolphosphoinositide anchors for membrane-tethering proteins. PMID- 11044997 TI - Fusions to members of fibroblast growth factor gene family to study nuclear translocation and nonclassic exocytosis. PMID- 11044998 TI - Posttranslational regulation of proteins by fusions to steroid-binding domains. PMID- 11044999 TI - Tet repressor-based system for regulated gene expression in eukaryotic cells: principles and advances. PMID- 11045000 TI - Coumermycin-induced dimerization of GyrB-containing fusion proteins. PMID- 11045001 TI - Use of glutathione S-transferase and break point cluster region protein as artificial dimerization domains to activate tyrosine kinases. PMID- 11045002 TI - Recombinant aequorin as tool for monitoring calcium concentration in subcellular compartments. PMID- 11045003 TI - Recombinant aequorin as reporter of changes in intracellular calcium in mammalian cells. PMID- 11045004 TI - Monitoring protein conformations and interactions by fluorescence resonance energy transfer between mutants of green fluorescent protein. PMID- 11045005 TI - Studies of signal transduction events using chimeras to green fluorescent protein. PMID- 11045006 TI - Use of fusions to green fluorescent protein in the detection of apoptosis. PMID- 11045007 TI - Synapto-pHluorins: chimeras between pH-sensitive mutants of green fluorescent protein and synaptic vesicle membrane proteins as reporters of neurotransmitter release. PMID- 11045008 TI - Studying organelle physiology with fusion protein-targeted avidin and fluorescent biotin conjugates. PMID- 11045009 TI - Fluorescent labeling of recombinant proteins in living cells with FlAsH. PMID- 11045010 TI - Ubiquitin fusion technique and its descendants. PMID- 11045011 TI - Use of phosphorylation site tags in proteins. PMID- 11045012 TI - NIH 3T3 cells or engineered NIH 3T3 cells stably expressing GDNF can protect primary dopaminergic neurons. AB - Glial cell line-derived neurotrophic factor (GDNF) shows potent and relatively specific protective effects on dopaminergic neurons. However, the size of the GDNF protein (MW 32-42 kDa) precludes the clinical use of GDNF via parenteral administration. It would thus be useful to have a cell line that stably secretes GDNF with full biological activities. The present study shows that NIH 3T3 cells express a considerable amount of GDNF. After co-culturing with primary E14-E16 midbrain neurons, such cells protected primary rat midbrain TH-immunopositive neurons from degeneration and MPP+ toxicity. In order to enhance endogenous GDNF expression, NIH 3T3 cells were stably transfected with GDNF cDNA with the Kozak sequence. The clones with the highest GDNF expression level were selected. The protective effects of engineered cells increased as the GDNF expression level increased. These cell lines may merit clinical investigation. PMID- 11045013 TI - Evaluation of outcome and QOL in patients with craniofacial resection for malignant tumors involving the anterior skull base. AB - The purpose of this study was to retrospectively evaluate the effectiveness of anterior craniofacial resection in the treatment of nasal and paranasal malignant tumors involving anterior skull base. Between 1992 and 1998, 13 patients with nasal or paranasal malignant tumors underwent this surgical procedure. The site and time of metastasis or recurrence, and survival outcome were retrospectively surveyed. Current status of long-surviving patients and their subjective assessment of the surgical treatment were also evaluated through questionnaires. Median follow-up period was 52 months. Nine patients (69%) were alive with no evidence of disease. Of these patients, eight had survived for more than three years. Recurrence or metastasis occurred in four patients (31%). The mean time interval between surgery and recurrence or metastasis was 11 months. According to the results of questionnaires to long-surviving patients, 89% patients had some complaints. In particular, complaints of unsightly appearance were manifested by all these patients. When the patients themselves evaluated their current conditions resulting from this surgical treatment, 63% were dissatisfied. These results suggest that this surgical treatment is valid for selected patients in regard to survival outcome. When the effectiveness of this treatment is evaluated, however, psychological and functional issues should not be taken lightly. PMID- 11045014 TI - Endoscopic carpal tunnel release surgery: report of patient satisfaction. AB - Recently, a trend has developed to use an endoscope to achieve carpal tunnel release. Proponents of the endoscopic technique believe it has benefits to patients that include minimal incision, minimal pain and scarring, a shortened recovery period and a high level of patient satisfaction. To test these beliefs, a retrospective analysis of the first 42 cases that were done between May 1997 and June 1998 was completed. Endoscopic carpal tunnel release surgery was performed on patients with the classical clinical and neurophysiological findings of carpal tunnel syndrome. The procedure was performed in an outpatient surgery center under primarily local anesthesia and by the same neurosurgeon (RG), who was blind to data analysis. The biportal technique (Instratek Inc., Houston, TX, USA) was used as described by Brown. The first 42 patients (n = 35, seven patients had bilateral surgeries) were sent a survey (modified Health Outcomes Carpal Tunnel Questionnaire, Health Outcomes, Bloomington, MN, USA) that measured a wide spectrum of variables, with a year follow-up. Patient demography indicated wide patient selection. All subjects (100%) had claimed work-related injury. Patient satisfaction was reported in 86%. No or mild incisional pain, night pain, absent tingling, and improved grip strengthening were reported in 100%, 95%, 81%, and 85% respectively. The mean for return to daily activity and work was 14 and 25 days respectively. No recurrent hematoma, infection, or structure injury was reported. Endoscopic carpal tunnel release can be done safely and effectively with excellent self-reports of patient satisfaction. Reduced recovery period and hospitalization with minimal tissue violation and incisional pain can be expected. PMID- 11045016 TI - Numbers of neurons and non-neuronal cells in human trigeminal ganglia. AB - We analyzed both trigeminal ganglia of eight post-mortem human subjects for their content of neurons and non-neuronal cells using dissociated cell suspension techniques. Neuronal counts in each of the 16 ganglia ranged from 20,000 to 35,400, with an average of 27,400 +/- 4800, and the estimated ratio of non neuronal to neuronal cells was 100 to 1. Our numbers are comparable to the lower ranges obtained using different techniques in previous studies. PMID- 11045015 TI - Activation of caspase-9 and -3 during H2O2-induced apoptosis of PC12 cells independent of ceramide formation. AB - The treatment of PC12 cells with H2O2 (100-500 microM) resulted in typical apoptotic changes including fragmentation and condensation of nuclei, and DNA fragmentation observed as DNA ladder. H2O2-induced apoptosis was associated with activation of caspase-3 as assessed by cleavage of specific fluorogenic substrate peptide and processing of procaspase-3 and poly(ADP-ribose) polymerase. However, formation of ceramide, which often locates upstream of caspase-3, was not observed. The inhibitory peptide relatively specific for caspase-3, z-DEVD-FMK and non-selective caspase inhibitor z-VAD-FMK inhibited activation of caspase-3 and apoptotic cell death. However, the relatively specific inhibitors, Ac-YVKD for caspase-1 and Ac-IETD for caspase-8/6, did not affect the occurrence of apoptotic cell death. As an upstream activation of caspase-3, induction of cytochrome c release followed by processing of procaspase-9 was observed by Western blotting, although the formation of intracellular ceramide was not observed. On the other hand, in PC12 cells overexpressing Bcl-2, the number of apoptotic cells was markedly decreased and activation of both caspases-9 and -3 was prevented. These results suggest that cytochrome c and caspase-9 initiate the activation of executor caspase-3 in H2O2-treated PC12 cells, and that Bcl-2 inhibits H2O2-induced release of cytochrome c from mitochondria and then proteolytic processing of procaspase-9. PMID- 11045017 TI - A case of spinocerebellar ataxia accompanied by severe involvement of the motor neuron system. AB - We report a sporadic case of spinocerebellar ataxia accompanied by later but severe involvement of the motor neuron system. A 72-year-old man began to show ataxia and dysarthria at age 66 years. Neurological examinations revealed saccadic eye movement, slurred speech, truncal ataxia, pyramidal sign, and urinary disturbance. Neither history of alcoholism nor hereditary factors were found. He developed muscular atrophy of the lower and upper extremities and limb ataxia within three years. Superficial and deep sensations were diminished in both feet four years after onset. Thus, he presented with cerebellar ataxia, bulbar sign, upper and lower motor neuron symptoms, sensory disturbance, and autonomic sign after six years at age 72. The level of serum, creatine phosphokinase (CPK) was increased, and muscle biopsy showed marked neurogenic change. Magnetic resonance imaging (MRI) revealed mild cerebellar and pontine atrophy. Although the combination of spinocerebellar ataxia and motor neuron disease is very rare, the present case suggests the inter-relation of the spinocerebellar and motor neuron systems, and presents peripheral neuropathy as a subtype of multisystem atrophy. PMID- 11045018 TI - Microsatellite instability in primary brain tumors. AB - Microsatellite instability has been reported in hereditary colorectal cancer syndrome and in various kinds of human sporadic tumors. It has also been shown in brain tumors, although with conflicting results. In the present study, DNA samples obtained from 20 primary brain tumors (10 glioblastomas, three astrocytomas, five meningiomas, one ependymoma, one hemangiopericytoma) were analyzed to detect microsatellite instability. Nine microsatellites, mono, di-, tri- and tetranucleotide repeat markers, located on nine different chromosomes, were used. Four of the 20 neoplasias (20%) showed microsatellite alterations in tumor DNA with respect to normal DNA. Two glioblastomas and one atypical meningioma (15%) showed additional bands or bands with shift of electrophoretic mobility, whereas allelic loss was observed in two glioblastomas (10%). In one glioblastoma, one allelic loss and one extra allele were observed at two different loci. These data indicate that in primary brain tumors there is not a high genetic instability. Although we used markers with inherently high levels of instability, only sporadic microsatellite alterations were found. Consequently, alterations in the mechanisms of DNA mismatch-repair, the most important cause of replication errors in hereditary and sporadic colorectal cancers, do not seem to play a major role in the oncogenesis of primary brain tumors. PMID- 11045019 TI - F-waves and facilitated late responses of the mentalis muscle in patients with a cerebrovascular accident. AB - F-waves in the extremities result from the backfiring of antidromically activated anterior horn cells and F-waves of the mentalis muscle can be also elicited after stimulation of the marginal mandibular branch of the facial nerve. In order to investigate the influence of the descending pathway of the excitability of the facial motonucleus, the F-wave of the mentalis muscle and the facilitated late response, which follows F-waves and which seems to be the snout reflex due to their similar latency and habituation, were studied in 11 conscious patients with a hemispheric cerebrovascular accident (CVA) presenting with hemiparesis, and in 10 unconscious patients with CVA or head injury. The duration and the persistence of the F-waves increased significantly statistically on the normal side in the CVA patients compared with those of the palsy side and the normal subjects. In comatose patients the F-waves and the facilitated late response were not elicited. The latency (46.1 +/- 13.3 msec) of the facilitated late responses in the unconscious patients tended to increase compared with the latency (36.6 +/- 4.3 msec) in the conscious patients. These findings suggest that the hyperexcitability of the facial motoneuron is ipsilateral to any hemispheric lesion; the hemispheric lesion exerts a bilateral excitatory influence on the interneuron of the facilitated late response: and that the reticular formation may influence the facial motoneuron and any interneurons concerned in the facilitated late response. F-waves and facilitated late responses should be further examined as neurophysiologically useful diagnostic methods. PMID- 11045020 TI - Spontaneous disappearance and reappearance of a ruptured cerebral aneurysm: one case found in a group of 33 consecutive patients with subarachnoid hemorrhage who underwent repeat angiography. AB - The spontaneous disappearance and reappearance of a ruptured cerebral aneurysm is generally assumed to be a rare phenomenon although the actual incidence is unknown. Among 39 consecutive cases of acute subarachnoid hemorrhage (SAH), 33 were studied by three-dimensional computed tomographic angiography (CTA) within 6 h after the onset of SAH, followed by digital subtraction angiography (DSA) within 24 h after the ictus. Of those patients, one, a 58-year-old woman, had a saccular aneurysm at the distal anterior cerebral artery; the aneurysm was clearly demonstrated by CTA 2.5 h after the SAH onset, but was not shown by a subsequent DSA performed 8.5 h after the ictus. A follow-up DSA detected the neck of aneurysm on day 11, and the whole aneurysm was visualized on day 19. The observations in this particular case suggest that the spontaneous disappearance of a ruptured cerebral aneurysm may occur during the ultra-early stage of SAH and that reappearance may follow during the next few weeks. The patient did not suffer complications such as vasospasm or systemic hypotension nor was she treated with antifibrinolytic agents. The aneurysmal shape and the surrounding clot are considered as putative factors possibly related to the intermittent appearance of the aneurysm. PMID- 11045021 TI - Bloody cerebrospinal fluid from patients with subarachnoid hemorrhage alters intracellular calcium regulation in cultured human vascular endothelial cells. AB - Endothelial cell dysfunction may contribute to cerebral vasospasm and aggravation of ischemic brain damage following subarachnoid hemorrhage (SAH). It has been suggested that oxyhemoglobin derived from subarachnoid blood clots might be a prime candidate for cerebral vasospasm. In this study, cisternal bloody cerebrospinal fluid (bCSF) was collected from SAH patients four and seven days after aneurysmal rupture, and the effects of bCSF on the cell growth and intracellular calcium ion ([Ca2+]i) dynamics were investigated in cultured human umbilical vein endothelial cells. CSF collected from patients undergoing other intracranial surgeries was used as a control. Pre-treatment with bCSF4 significantly facilitated cell proliferation and DNA synthesis in the cultured endothelial cells, and significantly enhanced histamine-induced [Ca2+]i increase, while acute treatment of the bCSF elicited no [Ca2+]i change. Pre-treatment with interleukin-1 beta showed a similar significant enhancement of the histamine induced [Ca2+]i response, while pre-treatment with high concentrations of serum or interleukin-6 did not change the [Ca2+]i response. It is concluded that bCSF collected from SAH patients contains some substances which enhance endothelial cell proliferation and sensitivity to inflammatory mediator. PMID- 11045022 TI - An unusual subdural empyema: case report. AB - Subdural empyema in a 38-year-old patient with congenital hemangioma, suppurative parotitisis, soft tissue phlegmonia and osteomyelitis is reported. The clinical, radiological and surgical features are outlined. A review of the literature reveals the uniqueness of this case. PMID- 11045023 TI - Extensive subdural empyema treated with drainage and barbiturate therapy under intracranial pressure monitoring: case report. AB - In subdural empyema (SDE), if the mass effect and vasogenic edema are not controlled, the brain can be fatally damaged. Massive SDE over the skull base often requires repeated surgical drainage for removal of accumulated pus. Intracranial pressure (ICP) management until obliteration of the empyema is important to the improvement of clinical outcome. An 18-year-old man was admitted to our center in a nearly comatose state and with a mild fever. CT scan showed massive SDE extending to the skull base and parafalx. ICP was measured with a pressure transducer through an intraventricle tube. Repeated surgical drainage was performed while ICP was controlled with barbiturate therapy. He was discharged with no neurological deficits. In patients with an extensive SDE over the cerebral hemisphere, ICP control with barbiturate therapy may enhance the therapeutic effect of surgical drainage. PMID- 11045024 TI - Transradial approach for selective cerebral angiography: technical note. AB - Selective cerebral angiography is currently being performed using transfemoral and transbrachial approaches. However, these techniques require patients to tolerate a prolonged focal compression and sometimes cause serious complications such as pulmonary embolism. The authors describe a technique of transradial approach as a safer selective cerebral angiography. Between July 1997 and November 1998, 70 patients underwent selective cerebral angiography with a transradial approach using a 4-F catheter. The collateral blood supply to the hand from the ulnar artery was confirmed using Allen's test prior to the procedure. To prevent the mechanical spasm of the radial artery, an arterial introducer 20 cm long was used. The radial artery was successfully punctured and cannulated in all patients. Selective catheterization of the intended vessels was obtained in over 98% of the carotid angiography and over 95% of the vertebral angiography. No major vascular complications such as cerebral infarction, upper limb ischemia, significant local hematoma or pseudoaneurysm were experienced. The transradial approach is a less invasive and safer technique for selective cerebral angiography, and could be an alternative to transfemoral and transbrachial approaches. PMID- 11045025 TI - An anatomical and pathological evaluation of middle cerebral artery occlusion in rats. AB - Adult male Sprague-Dawley rats (n = 87) weighing 350-400 g were used for studying the anatomy of the horizontal segment of middle cerebral artery and infarct area after occlusion of the artery. In the experimental group (n = 27) middle cerebral artery was coagulated 3-4 mm length from the origin of the lateral striate arteries to the inferior cerebral vein and divided. Control rats (n = 20) had all the surgical procedures except occlusion. Another group of rats (n = 40) were used to determine the anatomical variations of middle cerebral artery after intracarotid carbon black injection. Five major patterns of middle cerebral artery were observed and two of them were major and constituted 92.5% of rats. Twenty-four hours after middle cerebral artery occlusion, all animals were neurologically evaluated. On the third day after occlusion the brains were stained with 2%, 2,3,5-triphenyltetrozolium chloride. The area of infarction was assessed by computerized analysis method. In our study after determining the variations of the middle cerebral artery and its branches in our strain of rats, we were able to achieve 92.5% grade III and IV infarcted area. PMID- 11045026 TI - Investigation of blood flow in meningothelial and fibrous meningiomas by xenon enhanced CT scanning. AB - We investigated whether xenon-enhanced computed tomography was able to separate meningothelial meningioma from fibrous meningioma. Cerebral blood flow was studied by xenon-enhanced computed tomography in six patients with incidentally detected intracranial meningiomas. All of the tumors were small (< 32 mm) and there was little or no peritumoral edema. Three patients had meningothelial meningioma and three patients had fibrous meningioma. The tumor blood flow and the contralateral tissue blood flow were determined. The ratio of these parameters was 1.753 +/- 0.467 for meningothelial meningiomas and 0.809 +/- 0.105 for fibrous meningiomas, with a significant difference between the two tumor subtypes (p = 0.0185). There was no correlation between the signal intensity on magnetic resonance imaging and tumor subtype, and the findings on cerebral angiography also did not indicate the subtype. In conclusion, xenon-enhanced computed tomography showed a difference between smaller meningothelial and fibrous meningiomas in patients with normal surrounding brain tissue. We could not confirm that xenon-enhanced computed tomography was able to distinguish the subtype of meningioma because of the small number of subjects in this study, but our findings might expand interest in the clinical use of this method. PMID- 11045027 TI - Cerebral oxygen reactivity in the dog. AB - Brain tissue oxygen reactivity is a measure of the increase in tissue oxygen pressure (PtO2) relative to an increase in arterial oxygen pressure (PaO2). Clinical studies show that PtO2 reactivity is increased after cerebral injury. However, the impact of patient ventilation on these measures is not known. We determined whether changes in end tidal carbon dioxide pressure (ETCO2) would affect PtO2 reactivity in dogs. After a craniotomy, a Neurotrend probe that measures PtO2 was inserted into the cerebral cortex of eight dogs. PtO2 reactivity was measured at five concentrations of inspired oxygen (room air, 40%, 60%, 80%, 95%) at three levels of ETCO2 (20 mmHg, 40 mmHg, 60 mmHg) in random order. PtO2 reactivity at ETCO2 of 20 mmHg was 0.2 and increased to 0.3 when ETCO2 was 40 mmHg was 0.4 when ETCO2 was 60 mmHg (p < 0.05). These results show that PtO2 reactivity increases from hypocapnia to normocapnia. It is important to consider the ventilation state of each patient when evaluating PtO2. PMID- 11045028 TI - Metabolic and ionic responses to global brain ischemia in the newborn dog in vivo: II. Post-natal age aspects. AB - The main difference between newborn and adult brains is expressed in the relative resistance of the newborn brain to oxygen deprivation. The aim of the present study was to examine the effect of global ischemia in canine puppies of three different ages on the metabolic, ionic and electrical activity of the brain and to study the basic mechanisms underlying the relative resistance of the newborn brain in ischemic episode. The puppies were divided into three age groups. The young group included 0-6-day-old puppies (n = 16), the intermediate group included 7-19-day-old puppies (n = 21), and the 'adult' group included puppies aged 20 days or more (n = 17). Statistical analysis of the results led to the following conclusions: The younger the puppy, the longer is the time until the occurrence of the secondary reflectance increase SRI (13.0 +/- 1.9 min vs. 5.3 +/ 0.5 min). The younger the puppy, the longer the time until onset of potassium leakage from the cells (0.9 +/- 0.1 min vs. 0.35 +/- 0.05 min) and the lower the amount of potassium leakage (9.6 +/- 2.8 mM vs. 21.7 +/- 4.8 mM). The rate of pumping of the potassium ions into the cells during the recovery stage was higher in the oldest group (1.2 +/- 0.2 mM min-1 vs. 0.38 +/- 0.1 mM min-1). It was possible to speculate that in the young puppies there is uncoupling of the oxidative phosphorylation from respiration and as a result, there is a lower, if any, rate of ATP synthesis. It seems that the newborn brain is able to cope with a decrease in available energy for a longer period of time. This is apparently due to differences in membrane characteristics and an improved ability to retain ionic equilibrium across both sides of the membrane. PMID- 11045029 TI - Protective effect of FK506 in the reperfusion model after short-term occlusion of middle cerebral artery in the rat: assessment by autoradiography using [125I]PK 11195. AB - The protective effect of the immunosuppressant agent FK506 in the reperfusion after short-term occlusion of the middle cerebral artery in the rat model was evaluated using [125I]PK-11195 autoradiography. FK506 0.5 mg kg-1 day-1 was administered intramurally to Wistar rats weighing 260-300 g from one day prior to ischemia to seven days after ischemia. Reperfusion was performed after 30 or 60 min occlusion. Infarct area was evaluated by [125I]PK-11195 autoradiography on the seventh day following occlusion. FK506 significantly reduced the infarct area in the caudate nucleus following 30 and 60 min occlusion, but significantly reduced the infarct area in the cortex only following 60 min occlusion. These results suggest that FK506 has a protective effect against reperfusion after short-term occlusion of the middle cerebral artery. PMID- 11045030 TI - Prevention of cerebral vasospasm by nicardipine prolonged-release implants in dogs. AB - The purpose of this study was to determine the efficacy of nicardipine prolonged release implants for preventing vasospasm in a canine SAH model in a dose escalating placebo-controlled blind fashion. Drug-release kinetics of copoly(lactic/glycolic acid) pellet containing nicardipine were evaluated in vitro. In vivo, 18 dogs were randomly assigned to one of three groups, i.e. placebo, low-dose (0.8 mg), or high-dose (8 mg) nicardipine. Angiography was performed, followed by right craniectomy, the induction of subarachnoid hemorrhage, and the placement of the pellets in the Sylvian fissure. On Day 7 and Day 14, the angiography was repeated. In the first four days, 61.9% of the actual nicardipine loaded was released and within 10 days, 96%. The average percent reductions of vessel diameters in the middle cerebral artery on Day 7 were 43%, 14% and 7% in the placebo, low-dose, and high-dose groups, respectively (p = 0.0319). The mean concentration of nicardipine in the clots on Day 14 was 9.7 x 10(-7) mol-1 l-1 and 5.1 x 10(-6) mol-1 l-1 in the low-dose and high-dose group, respectively. This drug delivery system prevented vasospasm in dogs significantly even at low dose, while maintaining an appropriate concentration of nicardipine in the clot adjacent to the arteries. PMID- 11045031 TI - Comparison of brain tissue and local cerebral venous gas tensions and pH. AB - Neurosurgical monitoring devices have recently become available which are capable of measuring cerebral tissue gas tensions and pH. Brain tissue sensors have not been conclusively demonstrated to correlate with other measurements of regional cerebral gas tensions or pH. The present study was undertaken to correlate sensor values for pO2, pCO2 and pH with blood samples taken concurrently from local cerebral veins. Adult mongrel dogs were anesthetized and a craniotomy was performed. A small gyral vein was isolated and cannulated. Adjacent to the venous catheter tip, a Neurotrend brain tissue probe was inserted in an intracortical location. Each subject received a sequence of manipulations in inspired oxygen and end tidal carbon dioxide conditions. Under each experimental condition, samples of arterial and gyral venous blood were obtained and blood gas analysis performed. Concurrent brain probe measurements of tissue pO2, pCO2 and pH were recorded. Statistical analysis determined that local tissue and cerebral venous blood values for pO2, pCO2 and pH were highly correlated (R(s) = 0.62-0.82; p < 0.001). This indicates that there exists a confirmable monotonic relationship between tissue values and conditions in the post-capillary venous bed. Tissue sensors such as the Neurotrend probe can offer reliable trend indications in brain tissue gas tensions and pH. PMID- 11045032 TI - [Accidental morbidity in adolescence: a retrospective study in 12 to 15 year-old school children in Switzerland] . AB - Mortality and morbidity from injuries in the adolescent population is substantial in Switzerland as all over the world. The estimation of morbidity from injuries is difficult and depends on the design of the study. Little information is available on morbidity between ages 12 and 15. The Health Behaviour in School Aged Children (HBSC) study is an international survey based on a self administered questionnaire for which the Swiss Institute for Prevention of Alcohol and Drug Problems (SIPA) is responsible in Switzerland. 7196 schoolchildren aged 12 to 15 participated in the 1998 survey. Questions covered subjects such as health and health behaviour. A few questions covered injuries, with reference to frequency, context and consequences. 45.3% of adolescents have been treated at least once by a nurse or a doctor for an injury during the past 12 months. It can be deduced that nearly 149,000 adolescents aged between 12 and 15 are injured in more than 260,000 accidents in Switzerland. The rate of injury is 793 per 1000 adolescents per year. Furthermore, one adolescent out of ten injures himself three times or more per year. In conclusion, morbidity due to injuries in adolescents aged 12 to 15 in Switzerland is therefore considerable. This study demonstrates the high prevalence of repeated injuries in adolescents. PMID- 11045033 TI - [Nasal CPAP therapy in obstructive sleep apnea syndrome: patient compliance]. AB - We aimed in 186 patients with obstructive sleep apnoea, consecutively treated with nasal CPAP between January 1990 and December 1997, to evaluate compliance with nCPAP therapy and to explore factors influencing compliance. At 3 to 6 months (K1), 1 to 2 years (K2) and more than 3 years (K3) patients were reexamined regarding clinical conditions, the mean time of nCPAP usage per night, nCPAP pressure, and body mass index (BMI). The regularly conducted controls consisted of checking mask fitting, polygraphy with automatic pressure titration and once, mostly at K1, polysomnography. Compliance with nCPAP was considered to be sufficient more than 4 hours of usage per night. At K1, 9 patients had stopped therapy, 5 had moved away, 6 had changed therapy, 6 had died, and 41 had not been treated long enough to have a follow-up at K1. Thus we were able to reexamine 119 patients with ongoing nCPAP therapy. We found no correlation between indices of severeness of sleep apnoea (apnoea/hypopnea index, mean low of night time oxygen saturation, nCPAP pressure, and BMI) and compliance. At K1 87 patients (73%) had sufficient nCPAP compliance. All of them showed sufficient compliance at K2 and K3 too. We conclude that a sufficient compliance at an initial control implies sufficient compliance later on. 32 patients (27%) showed insufficient compliance at K1. Of this group 41% (13 patients) improved compliance at K2. This result underlines the value of a second instruction in nCPAP therapy when patient compliance was lacking initially. PMID- 11045034 TI - [Pheochromocytoma: diagnosis and treatment]. AB - Phaeochromocytoma is a rare tumour which produces a variety of symptoms. The most important factor is to think of the diagnosis, and there are many biochemical and pharmacological tests as well as radiological procedures to confirm it. Once the phaeochromocytoma is localised, it should, if possible, be removed. Surgery is the treatment of choice. In 1972 Ross described the diagnosis and therapy as "think of it, confirm it, find it and remove it". Today, 28 years later, this paper reviews the diagnosis and therapy of phaeochromocytoma under these key headings. PMID- 11045035 TI - [Helicobacter pylori and skin diseases--a (still) intact myth?]. AB - Helicobacter pylori plays a key role in the aetiology of peptic ulcer, gastric cancer and gastric MALT-lymphoma. Based on a number of reports, a possible relationship of Helicobacter pylori infection to a variety of different dermatoses has been suggested, including urticaria, rosacea, acne-rosacea, atopic dermatitis, alopecia areata, Sjogren's syndrome, Schonlein-Henoch purpura, and Sweet syndrome. Larger case-control studies, however, do not confirm this relationship. Therefore, Helicobacter pylori eradication therapy cannot be generally recommended in these dermatoses. PMID- 11045036 TI - [Hydropneumothorax in a patient with anorexia nervosa]. PMID- 11045037 TI - [The aging man]. PMID- 11045038 TI - [PADAM from the urologic viewpoint]. AB - PADAM stands for partial androgen deficiency in the aging male, and it is currently diagnosed with a testosterone level below 3 ng/ml (300 ng/dl or 12 nmol/l), and with symptoms varying according to the individual. The symptoms are a reduction or even loss of libido, a decline in muscle mass and strength, enhancement of visceral fatty tissue-padding, dryness of the skin, apathy, tiredness and distortion of mood right up to depression, and ostalgia due to osteoporosis. Before starting any form of hormonal substitution, which is only indicated if clinical symptoms and testosterone deficiency correlate, it is absolutely essential to exclude prostate cancer by using clinical evaluation and PSA values. Close PSA monitoring is necessary during testosterone substitution. In more than 95% of all patients with erectile dysfunction, the cause is not testosterone deficiency. Even a decreased level of dehydroepiandrosterone (DHEA) in an elderly male needs no replacement. There is also no indication for estradiol therapy in men--except in the rare case of aromatase deficiency. PMID- 11045039 TI - [The endocrine basis of sexual dysfunction in the elderly]. AB - There are no direct associations between testosterone levels and sexual function in the aging male. A study of 169 patients focusing on the relations of hormone levels and sexual dysfunction did not reveal associations between testosterone levels and the risk of altered sexual functions. In volunteers, in whom a hypogonadism was generated by the application of GnRH, no alterations in sexual reactions occurred. It is possible that other testosterone fractions are more meaningful than the total testosterone. However, all the known fractions correlate closely. The significance of a normal testosterone level for a normal erection has been rarely considered up to now. In an animal experiment, the intracavernous pressure, the density of alpha-receptors, and the PDE5 activity depended on normal testosterone levels. It is not known whether different testosterone levels influence the effectiveness of treatment procedures for erectile dysfunction in humans. The following questions with practical consequences have to be answered: Is the extensive determination of the bioactive testosterone necessary? Does the efficacy of treatment for erectile dysfunction depend on testosterone levels? PMID- 11045040 TI - [Age-related complaints and testosterone deficit from the psychosomatic viewpoint]. AB - Age-related changes in men resemble symptoms of hypogonadism. Although the average levels of testosterone decrease with age, correlations between complaints and testosterone levels are inconsistent and low in aging men. This is related to methodological limitations of studies, the high interindividual variability of testosterone in the aging male, ambiguities of normal values, and numerous determinants (e.g., health status, health behavior) on the level of testosterone. When studying the relationship between decreased testosterone, psychological and physical complaints have to take into consideration a whole array of psychosocial influences (e.g., perception, interpretation, and coping with age-related changes). PMID- 11045041 TI - [Testosterone and the prostate]. AB - The normal development and function of the prostate, as well as its pathological growth, are governed by a lifelong dependency on endogenous androgens, the majority of which are of testicular origin. In contrast to other androgen sensitive tissues, androgenic effects in the prostate are only exerted by the intracellular metabolite dihydrotestosterone. Conflicting evidence exists regarding changes of the intracellular prostatic androgen receptors in benign prostatic hyperplasia and prostatic carcinoma. Equally conflicting is the clinical evidence concerning androgen metabolism in patients with manifest or metastatic prostate cancer. So far, there is no clear evidence for an increased risk of prostatic neoplasia with testosterone substitution in absolute or partial androgen deficiency. However, in view of the high prevalence of latent prostate cancer, caution is advisable. PMID- 11045042 TI - [Testosterone substitution in the aging man]. AB - Hypogonadism is the main indication for testosterone substitution in the ageing male. The choice of the testosterone preparation should be based on the principle that serum levels of testosterone remain in the normal range and fast termination of therapy is possible. To date, transdermal testosterone systems appear to be best suited for substitution therapy in the ageing male. Future promising developments are testosterone gels and intramuscular testosterone undecanoate. Any testosterone therapy in the ageing male has to be monitored closely, especially regarding erythropoiesis and the prostate. PMID- 11045044 TI - [Significance of bacterial prostatic colonization for nosocomial urinary tract infections after transurethral prostate resection]. AB - Nosocomial urinary tract infections (UTI) are frequent complications after transurethral prostatectomy. The resection itself, postoperative catheterization and the prostate are possible causes of these infections. In this prospective study we investigated the influence of bacterial prostatic colonization on the incidence of postoperative urinary tract infections and inflammatory complications. In 78 patients we observed in 42 cases (53.8%) a bacterial prostatitis. In 14 patients (17.9%) we found nosocomial UTI's and in 12 patients (15.4%) inflammatory complications. The incidence of postoperative UTI's increase significantly in patients with positive prostate-cultures. On the other hand we only found corresponding prostate- and postoperative urine-cultures in less than 50%. The presented data are not sufficient to conclude the kind of relevance of bacterial prostatic colonization for postoperative UTI's in transurethral prostatectomy. Under consideration of the significant increase of nosocomial UTI's in patients with positive prostate cultures a perioperative antibiotic prophylaxis seems to be required in general. PMID- 11045043 TI - [Diagnostic and therapeutic consequences of kidney injuries in pediatric blunt abdominal trauma]. AB - Renal lesions are frequently encountered in blunt pediatric abdominal trauma. In this retrospective study, we analyzed the incidence of renal trauma in these trauma patients to determine which diagnostic and therapeutic approaches were most predictive. From 1976 to 1996, 308 children sustaining blunt abdominal trauma were admitted to our department. Patients were evaluated using abdominal paracentesis, ultrasonography, and urinary analysis. In specific cases, i.v. urography, CT scan, and/or angiography were applied. We used the Organ Injury Scale (OIS) for classification of renal trauma into five grades. We encountered 69 serious abdominal traumas. Thirty-six patients sustained renal lesions grade 2 (G2) or higher; 20 children were polytraumatized. There were 67 renal lesions with 28 G1, 22 G2, 8 G3, 5 G4, 1 G5, and 3 lesions of the urinary tract. Ultrasonography and urinary analysis were found to be optimal diagnostic means for screening and observing the course of renal lesions. For lesions G2 or higher, CT scan was the most reliable in classifying and diagnosing renal lesions. This superseded i.v. urography. If no contrast medium was excreted in the CT scan, angiography was indicated. Only ten patients proceeded to operative therapy. During the period reviewed, a shift from operative to conservative treatment was notable with a tendency toward minimally invasive therapy. If lesions were G4 or G5, operative treatment was always indicated. PMID- 11045045 TI - [Continent cutaneous urinary diversion of ileal neobladders by transverse tubularized ileum segments (Yang- or Monti-principle)]. AB - The method using transvers retubularized segment of ileum as an efferent conduit in Hautmann's neobladder is easy to perform and it can be repeated in revision procedures. We performed the procedure in nine patients who underwent radical cystectomy. At 8-16 months after operation, the resulting stomas were easy to catheterize. Of the nine patients, eight were continent. We find the procedure to be a good alternative when the appendix is absent and a cutaneous stoma is needed. It could be the preferred method in ileal neobladders with cutaneous outlet. PMID- 11045046 TI - [Postoperative function and aspects of the quality of life after Schroder-Essed technique of penile straightening]. AB - The Schroder-Essed technique is a standard technique for correction of penile curvature. Although it has been described as a simple and rapid technique offering low morbidity, no data are available on long-term follow-up and life quality of the operated patients. In a retrospective analysis we investigated functional results and life quality (LQ) using a detailed questionnaire. From January 1994 to January 1999, 40 patients (median age 26) underwent correction of penile curvature using a slightly modified Schroder-Essed technique. We used a self-established questionnaire investigating functional and cosmetic aspects and LQ. Complete follow-up data are available in 31/40 (77.5%) patients (congenital curvature, 19 patients; Peyronie's disease, 12 patients). Median follow-up is 22 months. Degree of angulation before surgery was estimated at < 45 degrees in 5 patients, 45-90 degrees in 22 patients, and > 90 degrees in 4 patients. In 21/31 patients (67.7%) cohabitation was not possible or discomfortable. Impaired LQ due to penile curvature was reported by 26/31 patients (83.8%). After surgery, the cosmetic result was considered as good or sufficient in 25 patients. Cohabitation was possible in 25/31 patients (80.6%). Penile shortening (> 2 cm) was reported by six patients. Significant improvement of life quality was reported by 15 patients. Among the 12 patients with Peyronie's disease, 6 mentioned postoperative impaired rigidity. In two patients with Peyronie's disease penile curvature recurred. This simple operation technique offers good functional and cosmetic results. Particularly patients with congenital curvature reported good or excellent results. Patients with Peyronie's disease have to be informed about the risk of disease recurrence. Other operation techniques should be considered for these patients. PMID- 11045047 TI - [Microhematuria--what next?]. AB - Asymptomatic microhematuria is a common reason for a urological consultation. Uncertainty prevails as to how meticulous the work-up must be, to not miss relevant or even life-threatening underlying diseases. To date, the Urological Associations have not released any guidelines to which extent patients need to be examined for asymptomatic microhematuria, which therefore is managed individually by each urologist. There are various potential examinations that can be applied, ranging from a clinical examination to a kidney biopsy. After reviewing the literature, an algorithm has been developed, which should assure diagnosis of serious disease and at the same time avoid costly, unpleasant and unnecessary examinations. PMID- 11045048 TI - [Expanded, radical perineal prostatectomy]. AB - One hundred and twenty-five consecutive patients with prostate cancer underwent an extended, radical perineal prostatectomy according to the technique described by VE Weldon. This technique was modified by an initial complete mobilization of the posterior aspect of the prostate and seminal vesicles from the rectum and pelvic wall, incision of the endopelvic fascia, and partial resection of the dorsal vein complex after suture ligature. The perioperative morbidity was low. An operative revision was necessary in four (3.2%) patients because of arterial bleeding from a drainage channel (n = 1), wound infection (n = 2), and rectocutaneous fistula (n = 1). The in-dwelling catheter was removed on day 4-8 in 104 (83%) patients. Positive surgical margins were diagnosed in 22 (17.6%) patients only. These patients had pT3 (n = 17) and pT4 (n = 5) tumors with a Gleason score > or = 7 (n = 17) mostly; extensive, multifocal capsular penetration (n = 18); seminal vesicle invasion (n = 11); and lymph node metastases (n = 4). The unifocal positive margins were localized at the apex (n = 3), dorsolateral (n = 6) aspect, and bladder neck (n = 4). In nine patients, multifocal positive surgical margins were noted. The risk for a positive surgical margin depends on the serum PSA level, Gleason score, and tumor volume. In case potency preservation is not considered, the extended, radical perineal prostatectomy with the above mentioned modifications should be considered to guarantee a low rate of surgical margins. PMID- 11045050 TI - [Aging male. Symposium 30 June-1 July 2000, Munich]. PMID- 11045049 TI - [Biomaterials in urology]. AB - Biomaterials are defined as non-living materials which are used in interaction with biological systems. Especially in the field of urology, biomaterials are applied in urinary diversion, urinary incontinence, erectile dysfunction, and as cosmetic prostheses. Biomaterial-tissue interaction is caused by the physical and chemical characteristics of the biomaterial, its degradation, and the resulting protein denaturation. General requirements include biocompatibility and functionality and the avoidance of carcinogenic, mutagenic, toxic, and allergic reactions. This is most important when there is permanent contact between urine and epithelial tissue, which may lead to biofilminfection and incrustation. Continuous modification of known materials, inauguration of new materials, as well as the possibilities of tissue engineering will determine their development in the years to come. PMID- 11045051 TI - [Management of patients with increasing PSA values after curative therapy]. PMID- 11045052 TI - [Guideline for after-care of patients with urinary diversion using intestinal segments. Guidelines of the German Society of Urology]. PMID- 11045053 TI - COX-2 inhibitors: a potential target for drug therapy in the management of colorectal cancer. AB - Colorectal cancer is currently the third most common cancer in Malaysia. Elevated expression of COX-2, an induced cyclooxygenase isoenzyme, has been seen in colonic adenomas and colorectal carcinoma. There is evidence that inhibition of this COX-2 can decrease the risk of colorectal cancer. Selective COX-2 inhibitors may have a role in reducing the risk of colorectal cancer in high-risk individuals. PMID- 11045054 TI - Amoebiasis: a 10 year retrospective study at the University Hospital, Kuala Lumpur. AB - A hospital based retrospective study of amoebiasis was carried out for a ten-year period at the University Hospital, Kuala Lumpur. Of the 51 cases traced, 30 (59%) had amoebic dysentery, 20 (39%) were amoebic liver abscess (ALA) and one patient had both conditions. Entameoba histolytica trophozoites were identified in 13 (43%) of the amoebic dysenteric stools and 9 (30%) from biopsy. Of the 20 (39%) ALA cases, only one showed parasites in the stool and biopsy. Majority of the patients with dysentery were Malays while Chinese comprised 40% with ALA. Males predominated overall with a male female ratio of 3:1, while for ALA it was 9:1. Most of ALA were single (71.4%) and were localised in the right lobe. The majority of the patients were unemployed. Eighty three percent (83%) of the patients presented with diarrhoea or dysentery followed by abdominal pain while those with ALA had fever, chills, rigors and pain in the right hypochondrium. Eighty percent of the ALA cases showed hepatomegaly. All patients responded to treatment with metronidazole. PMID- 11045055 TI - Deaths following acute diarrhoeal diseases among hospitalised infants in Kuala Lumpur. AB - The risk factors and modes of death following acute diarrhoeal illness in children admitted to University Hospital, Kuala Lumpur between 1982 and 1997 were studied retrospectively. Among 4,689 cases of acute gastroenteritis admitted, ten deaths were noted. The case mortality rate was 2.1/1000 admissions. All deaths were infants below one year, with eight females and two males. Acute renal failure and acute pulmonary oedema were common preceding events. Female sex, infants less than twelve months, the presence of hyper or hyponatraemia and moderate to severe dehydration on admission were risk factors for deaths. PMID- 11045056 TI - Demotivating factors among government doctors in Negeri Sembilan. AB - Motivation, especially on the relationship of remuneration of government doctors to it, has long been an issue of concern. This study sought to elucidate the demotivating factors in service and the perceived discrepancy in income. It was conducted amongst doctors serving in the Ministry of Health, Negeri Sembilan, using self-administered questionnaires. Factors considered demotivating were remuneration, workload and recognition given. Career development, promotion prospects, issues with superiors, resources and patient attitudes were other factors identified. On average, respondents expect an income of 1.63 times more than their current drawn salary and 87.2% cited rewards as a recommendation to improve their satisfaction in service. In-service training was desired by almost all. Though the medical profession has traditionally been viewed as altruistic in nature, doctors in service are voicing out their views and perception, and they should be heard. PMID- 11045058 TI - Revascularization for foot salvage in diabetic critical foot ischaemia. AB - A consecutive series of 32 diabetic patients, 16 male and 16 female, who presented to the authors with critical limb ischaemia was reviewed over a two year period. Atherosclerotic risk factors and co-morbidities were present in 56% of these patients. Diagnostic angiography was performed in all patients. Revascularization was achieved in 91% of the cases with three perioperative deaths. Ten bypasses were anastomosed distally to one of the crural or ankle arteries at the foot. Major amputations were required in five patients who had had revascularization and in 4 of these gross sepsis was the main factor responsible for limb loss despite patent grafts. The primary graft patency rates at one month and one year were 96% and 90% respectively. Surgical reconstruction was possible in the majority of diabetic patients with critical ischaemia and should be offered to patients preferably before the establishment of gross sepsis to improve limb salvage. PMID- 11045057 TI - The effect of pesticide on the activity of serum cholinesterase and current perception threshold on the paddy farmers in the Muda agricultural development area, MADA, Kedah, Malaysia. AB - BACKGROUND: The aim of this study was to find out the indicator as a marker of person who are exposed to pesticides (organophospate). Up to now the serum cholinesterase is used as a marker. This new method will not taking blood vein since this be the obstacle in conducting research in a rural area. METHODS: A cross sectional study was conducted to investigate the effect of pesticide exposure on the activity of serum cholinesterase (ChE) and current perception threshold (CPT). The samples are 60 paddy farmers (exposed group) and control group taken from the office of Muda agricultural area of Kedah in 19 do not expose to pesticide. The CPT values measured using a portable constant current electric nerve stimulator, Neurometer CPT/Eagle (Neurotron Incorporated, Baltimore, USA). Assessments carried out by one examiner on the index finger (median digital nerve) and the great toe (peroneal digital nerve). The current used are 3 neuroselective frequency range 2000 Hz, 250 Hz, and 5 Hz. The serum cholinesterase activity measured spectrophotometrically using cholinesterase inhibition test kit CHE MHE 1,144729. RESULT: It was significantly evident that the serum cholinesterase activity noted reduced among the paddy farmers (p = 0.014). The CPT values were significantly elevated for the 2000 Hz frequency range for both the measurement sites (index finger p < 0.0001 and great toe p < 0.0001). For the 250 Hz frequency range CPT values were significant only for the index finger (p = 0.012). However there was no significant difference for the 5 Hz frequency range. There was also a significant correlation (negative) between CPT values and serum cholinesterase activity more for the 2000 Hz frequency range (index finger r = 0.672, p < 0.0001 & for great toe r = 0.736, p < 0.0001). The results were suggestive of subclinical impairment of distal axonopathy considering the fact only 25% of the farmer showed clinical manifestation of numbness. CONCLUSION: The selective involvement of the large diameter sensory fibres (250 Hz and 2000 Hz) were reflective of toxic peripheral neuropathy. Neurometer CPT/Eagle used as indicator for detecting the effect of pesticide instead of using cholinesterase enzyme activity. PMID- 11045059 TI - Appropriateness of medical admissions from a Malaysian public primary care clinic. AB - Appropriateness of medical admissions from a Malaysian public primary care clinic (Outpatient Department, Hospital Ipoh) was assessed by two physicians using a modified appropriateness evaluation protocol. Of 122 admissions between 16/6/96 and 15/7/96, 107 records (88%) could be traced from the records office. Eighty percent (86/107) were found to be appropriate and 20% (21/107) inappropriate admissions. Inappropriate admissions included admissions to the wrong discipline and patients who could be investigated and stabilised as outpatients or could be referred to specialist clinics. Protocols, provisions for urgent referrals and medical updates for doctors are recommended. PMID- 11045060 TI - Markers of ventricular tachyarrythmias in patients with acromegaly. AB - INTRODUCTION: Sudden cardiac death is a known complication of acromegaly. Little is known of the exact mechanism leading to sudden cardiac death in these patients. Ventricular tachyarrhythmias may be an important cause. If this is so, clinical markers of ventricular tachyarrhythmias may be more common in this group of patients. The presence of these markers allow better risk stratification among acromegalic patients. METHODS: We performed signal averaged electrocardiography and analysed 12 lead electrocardiography for QT dispersion on 17 acromegaly patients who attended the UKM endocrine clinic within a period of 5 months and compared them with similar age matched controls. Signal averaged electrocardiogram was performed using Marquette Mac 12/15 ECG analyser and QT intervals were measured manually from 12 lead ECG tracings. Late potential positivity was defined by the standard Breithardt criteria. QT dispersion was defined as the longest minus the shortest QT interval from all 12 lead tracings. Echocariography was done to assess left ventricular hypertrophy in patients and controls. RESULTS: Late potential positivity was found to be more common in acromegaly patients compared with controls (chi-square, p < 0.05, n = 34) and QT dispersion was also found to be significantly higher in the acromegaly group compared with control s (mean +/- SE QT dispersion respectively 121.0 +/- 8.6 ms vs 86.2 +/- 7.0 ms, t-test, p < 0.05, n = 34). Left ventricular hypertrophy was present in five acromegaly patients and two in the control group. CONCLUSION: Acromegaly patients have a higher incidence of late potential positivity and higher QT dispersion compared with age matched controls. These findings might explain the increase susceptibility of these patients to sudden cardiac deaths from ventricular tachyarrhythmias. PMID- 11045061 TI - A comparison of the induction and emergence characteristics of sevoflurane and halothane in children. AB - This open labelled, randomised, controlled study was designed to compare the induction and recovery characteristics of sevoflurane and halothane anaesthesia in children. Forty American Society of Anaesthesiologist (ASA) physical status class 1 or 2 children (aged 1-10 year, weighed less than 25 kg) scheduled for elective urological procedure lasting less than one hour were allocated randomly to receive either sevoflurane (group S, n = 20) or halothane (group H, n = 20). The induction time in children receiving sevoflurane was significantly shorter than in those receiving halothane (mean (SD) 46 (13.6) second vs 69 (19.4) seconds, p < 0.005). The emergence from anaesthesia was also faster in children receiving sevoflurane than in those receiving halothane (mean (SD) 9 min (4.3 min) vs 21 min (8.9 min), p < 0.001). No major adverse effects were encountered in each group. We concluded that sevoflurane is comparable to halothane in Malaysian children. PMID- 11045062 TI - Hepatitis B surface antigen subtypes in hepatitis B seropositive subjects in University Hospital, Kuala Lumpur. AB - Hepatitis B surface antigen can be serologically defined as ayw1, ayw2, ayw3, ayw4, ayr, adw2, adw4 and adrq+ or adrq-. A study of common HBsAg subtypes in 44 HBsAg reactive sera in University Hospital was conducted using a solid-phase sandwich EIA. Eleven samples were found not typable and among the 33 typable HBsAg reactive sera, 3 HBsAg subtypes: adw, adr and ayw were identified. Subtype adw was found in 66.7% (22/33) of the typable HBsAg reactive sera; 24.2% (8/33) was of subtype adr and 6.0% (2/33) of subtype ayw. One sample was found to be reactive to both adw and adr. HBsAg subtype adw was found more commonly in Chinese but among the Malays, HBsAg subtype adr appeared to predominate. However, the small sample size precludes firm conclusions on the predominant subtype among the Malays. PMID- 11045063 TI - MRI appearances of peritoneal mesothelioma--a case report. AB - A rare case of diffuse malignant peritoneal mesothelioma in a 71 year-old Malay man with no previous history of asbestos or radiation exposure is described. The clinical manifestation was a large abdominal mass. At laparotomy he was found to be in the advanced stage of the disease. The tumour was not resectable and patient was sent home. He gradually deteriorated and died three months after diagnosis was made. The magnetic resonance imaging (MRI) features of peritoneal mesothelioma which has not been previously reported are described. PMID- 11045064 TI - Ileal strongyloidiasis in a Malaysian patient. PMID- 11045065 TI - Asbestosis in Malaysia: report on first two cases. AB - The first two cases of asbestosis in Malaysia are reported. Both had considerable occupational exposure to asbestos dust in the past, with a long latency period exceeding 30 years. One case presented with distinctive clinical and radiological features, while the other case was only confirmed by histological diagnosis. The usefulness of modern investigation techniques such as CT scan in the diagnosis of asbestosis is also illustrated. PMID- 11045066 TI - Scoliosis in a patient with lipodystrophy--treatment difficulties and literature review. AB - A 12 year-old Chinese schoolgirl presented with left-sided scoliosis at the age of 9 years. She has a rare defect in lipid metabolism, which is not known to be associated with spinal deformity. Her scoliotic curve deteriorated despite bracing. We report a rare occurrence of scoliosis in patient with lipodystrophy and the difficulty of using instrumented fusion in treating this condition. PMID- 11045067 TI - A successful pregnancy outcome in treated vulval rhabdomyosarcoma. AB - A 14 year old girl presented in 1986 with a huge perineal swelling which was progressively increasing in size and associated with loss of weight and loss of appetite. Biopsy from the mass revealed rhabdomyosarcoma of the vulva. She was treated with chemotherapy and radium implant. She responded well to the regime. Fibrosis of the vulva and vagina caused difficulty in consummation. Once it was corrected, she conceived easily and proceeded to a normal pregnancy and delivery. PMID- 11045068 TI - Massive bleeding from colonic diverticular disease with NSAID use. AB - Non-steriodal anti-inflammatory drugs (NSAID) are not only associated with bleeding in the stomach and duodenum, but can also complicate pre-existing diverticular disease of the colon. Here, a 58 year-old male with severe per rectal bleeding is presented and the role of NSAID as a causative factor of his problem is discussed. PMID- 11045069 TI - Respiratory diphtheria in three paediatric patients. AB - From August till November 1998, the Paediatric and Anaesthetic Units of Hospital Kuala Terengganu managed three patients from Kuala Terengganu District who were ventilated for respiratory diphtheria. Their ages were 5, 4 and 7 years old and their immunisation for diphtheria were not complete. All three patients presented with respiratory distress and were ventilated for upper airway obstruction. Their treatment included intravenous penicillin and diphtheria antitoxin. One patient died of cardiogenic shock with secondary pneumonia. Pharyngeal and tonsillar swabs of all three patients grew toxigenic Corynebacterium diphtheriae biotype mitis. There were 765 throat cultures taken from contacts. The confirmed positive cultures grew 2 toxigenic and 3 non-toxigenic Corynebacterium diphtheriae biotype mitis and surprisingly, 10 non-toxigenic biotype gravis. A prevalence study is needed to document the endemicity of diphtheria in Kuala Terengganu and to determine the carrier rate of both biotypes. Steps have been taken to increase the immunisation coverage in children. The giving of regular booster doses of diphtheria toxoid to the adult population should be considered. PMID- 11045070 TI - Cold agglutinins in low-grade B-cell lymphoma. AB - The presence of serum cold agglutinin can be the initial presentation of lymphoproliferative diseases. Conditions with persistent cold agglutinins are a spectrum of diseases that vary from benign lymphoproliferation of the "autoimmune like chronic cold agglutinin disease" to malignant lymphoma. We report a case of a 72-year-old woman who presented with severe anaemia, hepatosplenomegaly and episodes of peripheral haemagglutination precipitated by cold exposure. The haemoglobin was 5.6 g/dL with a cold agglutinin titer of 1:256 at 4 degrees C and 1:8 at room temperature (30 degrees C). The cold agglutinin showed anti-I specificity and kappa light chain restriction. Peripheral blood showed atypical lymphoid cells with a B-cell immunophenotype. Immunoglobulin gene rearrangement study by polymerase chain reaction (PCR) showed an amplified band at 100 bp, consistent with a clonal proliferation of B-lymphocytes. We believe that our patient had cold antibody haemolytic anaemia as the initial presentation of a low grade non-Hodgkin's lymphoma. The association of cold antibody haemolytic anaemia with low-grade B-cell lymphoma is unusual. PMID- 11045071 TI - Guidelines in the management of chronic obstructive pulmonary disease. A consensus statement of the Ministry of Health of Malaysia, Academy of Medicine of Malaysia and Malaysian Thoracic Society. PMID- 11045072 TI - Induction of apoptosis in neurogenic pulmonary edema. PMID- 11045073 TI - Credentialling in gastrointestinal endoscopy: recommendations of the Malaysian Society of Gastroenterology and Hepatology. PMID- 11045074 TI - Training programme in medical gastroenterology and hepatology: recommendations of the Malaysian Society of Gastroenterology and Hepatology. PMID- 11045075 TI - Changing concepts in lipid nutrition in health and disease. AB - Fat remains a hot topic because of concerns over associations between consumption of fats and the incidence of some chronic conditions including coronary artery disease, diabetes, cancer and obesity. Dietary fats serve multiple purposes. The effects of dietary fats generally reflect the collective influences of multiple fatty acids in the diet or food. This presentation highlights some recent developments on the role of dietary fats and oils in health and disease. Debate continues over the role of dietary modification in coronary prevention by lipid lowering. The degree to which a recommended diet will result in health benefits for an individual is difficult to predict, because the outcome will depend on the influence of other factors such as a person's genetic constitution, level of physical activity and total diet composition. There can now be little doubt about the importance of genetic factors in the etiology of cardiovascular disease, diabetes, obesity and cancer. The importance of antioxidant status in the prevention of cardiovascular disease as well as many cancers is being increasingly recognised. It is now evident that not all saturated fatty acids are equally cholesterolemic. Recent accounts evaluating palm oil's effects on blood lipids and lipoproteins suggest that diets incorporating palm oil as the major dietary fat do not raise plasma total and LDL cholesterol levels to the extent expected from its fatty acid composition. Palm oil is endowed with a good mixture of natural antioxidants and together with its balanced composition of the different classes of fatty acids, makes it a safe, stable and versatile edible oil with many positive health and nutritional attributes. In recent times, adverse health concerns from the consumption of trans fatty acids arising from hydrogenation of oils and fats have been the subject of much discussion and controversy. Trans fatty acids when compared with cis fatty acids or unhydrogenated fats have been shown to lower serum HDL cholesterol, raise serum LDL cholesterol and when substituted for saturated fatty acids, increase lipoprotein Lp (a) level, an independent risk factor for the development of coronary heart disease. The idea of which foods, nutrients and supplements are "healthy" is often being amended as new scientific data is presented and then simplified for the consumers. What was once perceived as a healthy diet is often no longer considered as such and vice versa. Dietary recommendations have to change with time and the evidence available. Nutritional recommendations should encourage eating a great variety of nutrient sources within our food supply in moderation. Various lifestyle options to improve health should also be promoted. PMID- 11045076 TI - Detecting meningococcal meningitis epidemics in highly-endemic African countries. PMID- 11045077 TI - Malaria interventions. PMID- 11045078 TI - The college links with St Paul's Cathedral. PMID- 11045079 TI - Medicine in the millennium: confronting the issues. PMID- 11045080 TI - Personality assessment techniques and ability testing as aids to the selection of surgical trainees. PMID- 11045081 TI - Home and away. PMID- 11045082 TI - Improving discrepancies: SpRs and consultant posts. PMID- 11045083 TI - The national plan for health. Patient Liaison Group. PMID- 11045084 TI - Education--who pays? PMID- 11045085 TI - Education versus service--the resident's dilemma? PMID- 11045086 TI - Lung volume reduction surgery: evidence for the resurrection of an old operation? PMID- 11045087 TI - Soft-tissue images. Hydatid disease of the liver. PMID- 11045088 TI - Musculoskeletal images. Soft-tissue mass at the site of a previous total knee arthroplasty. PMID- 11045089 TI - Musculoskeletal case 12. Presentation. Neurofibromatosis type 1. PMID- 11045090 TI - Soft-tissue case 35. Presentation. Cecal diverticulitis. PMID- 11045091 TI - Helicobacter and disease: still more questions than answers. AB - Since the causative role of Helicobacter pylori in peptic ulcer and gastritis was established, a number of advances have been made. Helicobacter virulence factors have been identified, the changes it causes in gastric acid secretion has been elucidated, and the entire genome of H. pylori has been mapped. Multiple lines of evidence indicate a strong link between the bacterium and noncardia gastric cancer. The infection can be confidently diagnosed by noninvasive serologic tests and the urea breath test. Triple therapy is almost always curative, and the infection almost never recurs in Canadian adults, but eradicating the bacteria in the absence of peptic ulcer only rarely leads to resolution of dyspepsia. New studies suggest that treating Helicobacter may increase the risk of peptic esophagitis and adenocarcinoma of the esophagus and cardia. PMID- 11045092 TI - The future of surgery--Santayana or Ford. AB - The Canadian health care system is currently in an era of reform and restructuring. Economic and political forces, changes in licensing and educational system as well as public expectations all influence change in this evolving health care delivery system. In contemplating change it is useful to remember the lessons of our rich surgical history so that the mistakes of the past will not be repeated. The Canadian Association of General Surgeons is well positioned to exert a leadership role in the evolution of surgical care in Canada. The role of the Association in the promotion of evidence-based surgery, continuing professional development and the provision of surgical services in rural areas is discussed in this paper. PMID- 11045093 TI - Small-bowel obstruction secondary to malignant disease: an 11-year audit. AB - OBJECTIVE: To determine the efficacy and long-term prognosis for operative versus nonoperative treatment of small-bowel obstruction (SBO) secondary to malignant disease. DESIGN: A chart review. SETTING: A university-affiliated teaching hospital. PATIENTS: The medical records of all patients with malignant disease as the established etiology of their obstruction who presented to the Sir Mortimer B. Davis-Jewish General Hospital, Montreal, between 1986 and 1996 were reviewed. There were 32 patients accounting for 74 admissions. INTERVENTIONS: Selective nonoperative management and exploratory laparotomy, immediate or delayed. MAIN OUTCOME MEASURES: The value of nonoperative management and need for operation. RESULTS: Colorectal and ovarian neoplasms were the principal primary malignant diseases that led to SBO. The median time between diagnosis of the malignant disease and SBO was 1.1 years. At their initial presentation, 80% of patients were treated by operation, but 47% of these patients had an initial trial of nonoperative treatment. Reobstruction occurred in 57% of patients who were operated on compared with 72% of patients who were not. The median time to reobstruction was 17 months for patients who underwent operation compared with 2.5 months for patients who did not. Also, 71% of patients were alive and symptom free 30 days after discharge from operative treatment compared with 52% after nonoperative treatment. Postoperative morbidity was 67%. Mortality was 13%, and 94% of patients eventually died from complications of their primary disease. CONCLUSIONS: SBO secondary to malignant disease usually indicates a grim prognosis. Operative treatment has better outcome than nonoperative management in terms of symptom free interval and reobstruction rates. However, it is marked by high postoperative morbidity. We recommend that, after short trial of nasogastric decompression, patients with obstruction secondary to malignant disease be operated on if clinical factors indicate they they will survive the operation. PMID- 11045094 TI - Blood flow changes to the proximal femur during total hip arthroplasty. AB - OBJECTIVE: To determine the changes in perfusion to the proximal femur that occur during cemented and uncemented total hip arthroplasty (THA). DESIGN: Case series. SETTING: A single tertiary-care centre. PATIENTS: Twenty-two consecutive patients. Those who had undergone previous hip surgery or received systemic corticosteroid therapy were excluded. INTERVENTION: Cemented (11 procedures) or uncemented (12 procedures) THA. MAIN OUTCOME MEASURE: Changes in blood flow at the level of the proximal femur, measured with laser Doppler flowmetry at 4 different times during THA. RESULTS: In both the cemented and the uncemented procedure overall proximal femoral blood flow was reduced (p = 0.002, p = 0.008, respectively). A greater reduction in overall proximal femoral perfusion was seen in the cemented group compared with the uncemented group (p = 0.004). This greater reduction in perfusion was seen primarily in the proximal femoral diaphysis (p = 0.004). CONCLUSION: The extensive canal preparation involved with the cemented procedure or the introduction of bone cement under pressure into the femoral canal may contribute to the greater reduction in perfusion to the proximal femur. PMID- 11045095 TI - Properties of a hybrid plaster-fibreglass cast. AB - OBJECTIVE: To examine the suitability of a plaster-fibreglass hybrid cast for orthopedic applications, comparing them to plaster of Paris (POP) and fibreglass constructs. METHOD: Groups of 10 standardized hybrid, POP and fibreglass casts were studied. An Instron servohydraulic system was used to test the casts in 3 point bending and shear. OUTCOME MEASURES: Strength, stiffness, weight, thickness and cost of the 3 types of cast, and shear strength at the interface between the POP and fibreglass in the hybrid casts. RESULTS: The hybrid casts were twice as strong as the POP constructs, were stiffer and weighed 14% less but were thicker and cost 2.5 times more. They were almost as strong as and less than half the cost of the fibreglass constructs but were thicker, not as stiff, and weighed 42% more. The shear strength of the POP-fibreglass interface in the hybrid casts was higher than the 3-point bending strength of this construct by a factor of 3. CONCLUSIONS: Plaster-fibreglass hybrid casts should be considered for orthopedic use on the basis of their strength, stiffness, weight and cost, combined with their acknowledged advantages of good moulding ability and water resistance. PMID- 11045096 TI - Lung volume reduction surgery for the treatment of severe emphysema: a study in a single Canadian institution. AB - OBJECTIVE: To evaluate lung volume reduction surgery (LVRS) and its effectiveness in improving pulmonary function, exercise capacity and quality of life in a population of emphysema patients referred to and screened in a single centre. DESIGN: A prospective case series. SETTING: A Canadian tertiary care hospital. PATIENTS: Patients with severe emphysema, significant dyspnea and impaired exercise capacity interfering with quality of life. INTERVENTIONS: Bilateral LVRS was performed through a median sternotomy. MAIN OUTCOME MEASURES: Pulmonary function tests (preoperative forced expiratory volume in the first second [FEV1], residual volume [RV]), 6-minute walk (6 MW) distance, quality of life (Medical Outcomes Study 36-item short-form health survey) and degree of dyspnea (Medical Research Council of Great Britain dyspnea scale and the baseline and transitional dyspnea indices) were assessed before LVRS and at 6 and 12 months after. RESULTS: Fifty-seven patients were assessed for LVRS, of whom 10 were selected for surgery. Homogeneous distribution of disease was the most common reason for exclusion. Of the 10 patients operated upon, 1 died of acute cor pulmonale on the fourth postoperative day and 1 died of recurrent exacerbations of chronic obstructive pulmonary disease and chronic respiratory failure at 315 days postoperatively. In the surviving patients, the mean preoperative FEV1 increased from 0.70 L before surgery to 0.90 L at 1 year, with a mean relative increase of 33.4%. The mean RV decreased from 5.57 L to 4.10 L, with a mean relative decrease of 27.6%. The 6 MW distance increased from 302.7 m to 356.9 m at 1 year, with a mean relative increase of 21.6%. Quality of life and degree of dyspnea were improved significantly at 1 year after LVRS. Of the 5 patients on oxygen at home before surgery, 4 were able to reduce their requirements but not to discontinue oxygen. CONCLUSIONS: LVRS is an effective palliative treatment for dyspnea and poor exercise tolerance in highly selected patients. Although the duration of palliation is unknown, our results show that improvements in pulmonary function, exercise, quality of life and degree of dyspnea are preserved over the first year. Only a minority of the patients screened were eligible for surgery. The 2 deaths in our series emphasize the need for even further delineation of selection criteria. PMID- 11045097 TI - Lung volume reduction surgery: results of a Canadian pilot study. Canadian Lung Volume Reduction Surgery Study Group. AB - OBJECTIVE: To present preliminary experience with lung volume reduction surgery (LVRS) before the institution of the Canadian LVRS trial. DESIGN: A prospective case series between December 1995 and January 1997. SETTING: University hospitals in London and Hamilton, Ont. PATIENTS: Forty-nine patients who had disabling dyspnea or emphysema with hyperinflation, able to participate in respiratory rehabilitation. Twenty-three patients were excluded because of comorbid conditions precluding surgery, pulmonary hypertension, excessive steroid dependence, malnutrition, obesity, previous thoracotomy, large solitary bullae, concurrent malignant disease, chronic bronchitis, hypercapnia or psychiatric illness. INTERVENTIONS: Preoperative respiratory rehabilitation followed by LVRS via median sternotomy. MAIN OUTCOME MEASURES: Impairment, disability and handicap were assessed before and 12 months after LVRS. Impairment was assessed by changes in pulmonary function test results and blood gas measurements, disability by the 6-minute walk test and cardiopulmonary exercise test, and handicap by the disease specific chronic respiratory disease questionnaire (CRQ), the generic medical outcomes survey short form 36 (SF-36) and the generic health utilities index mark III (HUI-III). RESULTS: Two patients died of respiratory failure while in rehabilitation. Twenty-four patients (17 men, 7 women) successfully completed rehabilitation and underwent LVRS. The mean age was 63 years (range from 49 to 78 years) and the median length of hospital stay was 12.5 days (range from 7 to 90 days). Two patients (8%) died in the early postoperative period (within 30 days) of pneumonia. One patient died of respiratory failure 8 months after LVRS after a difficult 90-day postoperative hospital stay. There were 27 major complications. There was a 36% relative increase in the mean forced expiratory volume in the first second (p = 0.01) and a 10% relative increase in the 6-minute walk test (p = 0.06). The mean CRQ dyspnea score increased 2.3 points (p = 0.01), and the SF 36 general health domain increased 20 points (p = 0.01). There was no significant change in the HUI-III (p = 0.73). CONCLUSION: LVRS appears to lessen the respiratory impairment and handicap for at least 1 year in selected patients with advanced emphysema. PMID- 11045098 TI - Canadian Network for International Surgery: development activities and strategies. AB - The Canadian Network for International Surgery (CNIS) is a surgical development and research organization, whose objective is to reduce death and disability from surgical disorders in low income countries. The organization has 4 main activities: (1) the Essential Surgical Skills (ESS) program teaches surgery to general practitioners and is predicated on the assumption that there will not be enough surgeons in Africa in the foreseeable future and therefore nonsurgeons must do surgery; (2) the injury control program, which is predicated on the conclusion that the incidence of injury in Africa is unacceptably high, therefore injury prevention is an imperative surgical strategy; (3) the library project, which sends new and recent books and journals to the surgical libraries of our African partners; and (4) the members' projects, which encourage individual or organization members to use their own creativity in meeting CNIS objectives. The CNIS has direct activity in 4 African countries and presents its project check list as a means to help others succeed. Canadian surgical and allied specialists can help in the reduction of needless suffering by supporting the CNIS. PMID- 11045099 TI - Perioperative transfusion or albumin as risk factors in colorectal surgery. PMID- 11045100 TI - Health care funding in surgical practice and the Canadian health care system. PMID- 11045101 TI - Health care funding in surgical practice and the Canadian health care system. PMID- 11045102 TI - Job satisfaction and dissatisfaction in Hungary: nurses' opinion of their profession in a changing society. PMID- 11045103 TI - The patient as CEO. Passion in practice. PMID- 11045104 TI - Hospital restructuring: does it adversely affect care and outcomes? AB - The past decade has witnessed pronounced changes in the organization of United States hospitals, many the direct result of restructuring and re-engineering initiatives intended to decrease costs and increase productivity. Little is known about how these initiatives have affected clinical care and patient outcomes. Using data from a variety of sources, the authors describe initiatives that hospitals undertook during this period, discuss how nurse staffing changed relative to the case mix of patients receiving care, and examine changes in nursing practice environments from 1986 to 1998. PMID- 11045105 TI - Building community in the healthcare workplace, Part 3: Belonging and satisfaction at work. AB - Squeezed between mounting budget pressures and staffing demands, healthcare managers have little room to maneuver as they are buffeted by incessant new strategies. So what is the constant that they can use to keep a steady course through all the chaos? We suggest the overarching constant is "community in the workplace." Although community building may have both short- and long-term benefits, it is not a strategy but a career-long philosophy to help managers tie together all the facets of their daily work into a more meaningful and satisfying purpose. This article, part 3 in a 4-part series, describes methods for building community in the entire organization or in a single unit. Options for implementing change in complex adaptive systems such as hospitals are discussed, along with remedies for the most common obstacles managers will encounter. Part 1 (July/August) described the negative dynamics that obstruct sustainable change and reasons for moving to community at work. Part 2 (September) discussed the basic characteristics and principles of community building. The final article (November) will present a case study and will discuss how community can be built in a unionized environment. PMID- 11045106 TI - Factors influencing outcomes after delegation to unlicensed assistive personnel. AB - The authors discuss a national survey of licensed nurses that describes factors associated with patient outcomes when nursing activities are delegated to unlicensed assistive personnel (UAP). Licensed nurses' overall experience and UAP's experience in the current work setting were associated with more positive events. When the outcome of the delegated activity was determined by routine observation, more positive events occurred. However, when there was no direct supervision of the UAP, more negative events occurred. Recognition of the importance of the supervisory process has implications for educational opportunities that focus on strengthening licensed nurses' delegation competencies. PMID- 11045107 TI - Analysis of the nursing work force compared with national trends. AB - Nursing work force analysis gives crucial data for administrators, allowing them to anticipate employment opportunities and areas of limited growth. Using an economic model of supply and demand, this study examined the employment and educational needs for nurses. In many areas of data collection, the District of Columbia data paralleled national trends, indicating the nationwide relevance of the conclusions. Projected mismatches between the type of nurses (by educational preparation) entering the work force and the skills required for the job vacancies could lead to increased demand for nurses with certain credentials as well as shortages of nurses in certain types of facilities. PMID- 11045108 TI - The professional nurse education program: a work force development model. AB - Invariably, healthcare institutions respond to repeated voids in professional nurse positions by recruiting persons external to the organization, rather than examining their nonprofessional ranks for high achievers worthy of formal career advancement opportunities. The authors describe why and how their professional work force development operates, the demographics of participants, and outcomes, such as retention rate, gains in college preparatory competencies, changes in self-reported psychosocial aspects of participants, influence of work hours on academic achievement, and financial investment. PMID- 11045109 TI - Sexual harassment in the workplace: nurses' perceptions. AB - This study examined the perceptions of nurses and nursing administrators of various types of sexual harassment. It also investigated the differences in the perceptions based on gender, previous harassment, and position. Significant differences in perceptions of sexual harassment were found according to types of harassment, gender, position, and sensitivity from prior harassment. These results can inform the educators and supervisors about where the message about harassment is not getting through strongly enough, and help them to focus their training efforts. PMID- 11045110 TI - [Augmentin and respiratory tract infections in the adult: current aspects]. PMID- 11045111 TI - [Why is the augmentin dose changing?]. PMID- 11045112 TI - [Predictive pharmacodynamic criteria and efficacy of augmentin 1 g/125 mg in 2 doses per day]. PMID- 11045113 TI - [Augmentin 1 g/125 mg 2 times a day in acute exacerbations of chronic bronchitis]. PMID- 11045114 TI - [Augmentin 1 g/125 mg 2 times a day in acute maxillary sinusitis]. PMID- 11045115 TI - [Sensitivity to antibiotics of bacteria from nosocomial infections. Evolution in resuscitation services of military hospitals]. AB - OBJECTIVE: The aim of this study, conducted in the French Military hospitals, was to monitor the course of the antimicrobial sensibility of bacteria isolated from nosocomial infection in intensive care units. PATIENTS AND METHODS: A prospective study has been conducted from January to December 1998 in all the intensive care units of the French Army. All the non-repetitive strains isolated from nosocomial infection were collected and sent to a reference centre. Antimicrobial susceptibility was determined by the agar dilution method. Beta-lactamase were identified by iso-electro-focalisation. Antibiotics choice and interpretative criteria were those of the "Comite Francais de l'Antibiogramme de la Societe Francaise de Microbiologie". RESULTS: A total of 849 strains are included in this study. Pseudomonas aeruginosa was the most frequently isolated bacterium (20%) followed by Escherichia coli (19%) Staphylococcus aureus (15%), coagulase negative Staphylococci (CoNS) (11%) and Enterococci (7%). Imipenem was the most effective antibiotic against enterobacteriaceae (336 isolates; 100% susceptibility). Gentamicin (92%), amikacin (92%) third generation cephalosporins (83%), aztreonam (83%) and ciprofloxacin (78%) were also very effective. Resistance to III generation cephalosporins was correlated with an extended spectrum beta-lactamase (BLSE) in 36% of cases. This BLSE could be associated with an over production of the constitutive cephalosporinase. The most frequent species producing BLSE were Enterobacter aerogenes (75% of BLSE) and Klebsiella pneumoniae (17%). Among the 172 P. aeruginosa isolated, antimicrobial susceptibility were respectively: 71% for imipenem, 62%: tobramycin, 60%: amikacin 59%: ciprofloxacin 59% piperacillin + tazobactam, 55% piperacillin, 53%: ceftazidime and 44% for ticarcillin. Seventy per cent of the 96 CoNS and 50.2% of the 126 S. aureus isolated were resistant to methicillin. A strain of S. aureus and 2 CoNS strains had intermediate resistance to teicoplanin. Twenty per cent of the 59 Enterococci strains isolated were resistant to aminopenicillins (10/11 strains of E. faecium), and 9% presented a high level of resistance to gentamicine. One strain of E. faecium was resistant to vancomycin. CONCLUSION: The evolution of the susceptibility to antibiotics in intensive care units reflects the antibiotic pressure and level of cross-transmission. High rates of meticillin-resistance among staphylococci, of resistance to beta-lactams antibiotics among P. aeruginosa and of ciprofloxacin among Enterobacteriaceae are shown in this study. The implementation of appropriate strategies for surveillance and prevention is necessary. PMID- 11045116 TI - [Treatment of perforated duodenal ulcer by laparoscopy. 35 cases]. AB - OBJECTIVE: Laparoscopic treatment of perforated duodenal ulcer is an alternative to laparotomy. We reviewed our experience to determine feasibility and reliability of the surgical procedure and the subsequent morbidity and mortality. PATIENTS AND METHODS: This was a retrospective series of 35 patients who underwent laparoscopic repair of perforated duodenal ulcers between January 1994 and November 1999. The perforation was closed by interrupted sutures in 86% of cases associated with irrigation-suction of the abdominal cavity. RESULTS: Two complications were observed: pyloric stenosis in one patient with a large perforation and Douglas pouch abscess in another. Conversion to laparotomy was necessary in 8 cases: in 2 of them the edges of the perforation were fragile, in 3 the perforation could not be identified (posterior position), and in 3 intraabdominal adhesions were important. There were no deaths. The 2-year results are quite satisfactory. CONCLUSION: This study shows that the laparoscopic procedure is reliable and adapted to treat ulcer perforation if the size is less than 1 cm. PMID- 11045117 TI - [Hypocomplementemic urticarial vasculitis]. AB - BACKGROUND: Low-complement urticarial vasculitis is an uncommon condition associating urticaria, glomerulonephritis, obstructive ventilatory disorders, and anti-Ciq antibodies. CASE REPORT: We report a case in a 34-year-old woman who developed urticaria with purpura, membranoproliferative glomerulonephritis (creatinine 238 mumol/l) and bronchial obstruction with bronchectasia. Total complement and the C3 fraction were low. Anti-C1q antibodies were found in the serum and anti-DNA antibodies were negative. Aggravation of the respiratory and renal failure progressed despite corticosteroid therapy, leading to death at 4 months. DISCUSSION: Bronchial obstruction in low-complement urticarial vasculitis results from emphysema and is often life-threatening. Our case exhibited an unusual feature due to the lack of radiodetectable emphysema, the presence of bronchectasia and the rapid degradation of the respiratory function. PMID- 11045118 TI - [Prevalence of hypertransaminasemia in untreated celiac disease]. PMID- 11045119 TI - [Thrombopenia in treatment with mesalazine]. PMID- 11045120 TI - [Schoenlein-Henoch purpura in Lyme disease]. PMID- 11045121 TI - [Increased dosage of inhaled corticosteroids or addition of salmeterol in symptomatic asthma]. PMID- 11045122 TI - [Apropos of hemocultures in cancerology]. PMID- 11045123 TI - [Hematology in the elderly patient: is there progress? 42nd Congress of the French National Society of Internal Medicine in Provence--8-10 June 2000]. PMID- 11045124 TI - [Dermatologic manifestations of atrophic polychondritis]. AB - A FREQUENT OCCURRENCE: Dermatological manifestations are frequently observed in patients with relapsing polychondritis. However, they have not been extensively studied as polychondritis is a rare disease and the reported dermatological manifestations appear to be nonspecific. A WIDE SPECTRUM: The most frequently observed manifestations are aphthosis with sometimes complex aphthosis, nodular papular or purpuric lesions. Neutrophilic dermatosis has been reported in isolated cases. Histologically, these lesions correspond to nonspecific inflammatory infiltrates, vasculitis or thrombosis. Various pathological findings are observed in nodular lesions. RELATED AND OVERLAP CONDITIONS: Similar dermatological manifestations occur in Behcet's syndrome and in inflammatory bowel diseases. Patients with relapsing polychondritis and myelodysplasia very frequently have dermatological manifestations. Repeated blood cell determinations are therefore necessary in patients with relapsing polychondritis and dermatological manifestations. PMID- 11045125 TI - [Chronic electric stimulation of the internal globus pallidus and subthalamic nucleus in Parkinson disease]. AB - PATHOPHYSIOLOGY: In Parkinson's disease, the neurodegenerative process of the nigrostriatal dopaminergic pathways induces an increase in activity of the subthalamic nucleus and the medial globus pallidus, which cause inhibition of thalamo-cortical outputs explaining parkinsonism. HIGH-FREQUENCY DEEP BRAIN STIMULATION: The adverse effects induced by lesions of subcortical structures (thalamotomy, pallidotomy) have increased interest in chronic electrical stimulation proposed as a new therapy in Parkinson's disease. This technique is reversible and can be modulated with less adverse effects. TWO TARGETS: Two targets may be proposed in case of severe motor fluctuations: the medial globus pallidus and the subthalamic nucleus. Pallidial stimulation improves dramatically levodopa-induced dyskinesia and, with a variable degree, the parkinsonian triad. Subthalamic stimulation rapidly reverses akinesia, rigidity and tremor and also dyskinesias which progressively tend to diminish after decreasing L-dopa dosage. LONG-TERM EFFICACY: A follow-up period of a few years has confirmed that the beneficial effect is maintained. However, stimulation dose not prevent the development of certain symptoms (postural impairment, cognitive decline). LIMITED INDICATIONS: Chronic electrical stimulation of medial globus pallidus and subthalamic nucleus may be proposed for parkinsonian patients with severe motor fluctuations associated with abnormal involuntary movements which are not controlled by different medical therapies. Parkinsonian symptoms must still be levodopa responsive and cause severe clinical disability severely limiting daily living activities. Cognitive impairment and other severe pathologies are contraindications. PMID- 11045126 TI - [Standards, options and recommendations for good practice in hemoculture in cancerology]. AB - Excepting emergency and aplasia: two to three blood samples should be draw for culture an hour apart within a 24 period (standard). For emergency or aplasia: two to three blood samples should be drawn for culture before initiating early antibiotic therapy. The delay between samples drawn from different sites should be less than one hour (standard). For patients on antibiotics: four to six blood samples should be drawn for culture within 48 hours, outside ongoing antibiotic administration. If the patient is given corticosteroids, it is recommended to draw two or three blood samples in case of deterioration (agreement of the experts). Rigorous aseptic techniques must be used (standard). Culture media are chosen according to the institution's microbial ecology (standard). The volume of blood drawn should be adapted to the system used (standard). Culture positivity is determined at 24 to 48 hours. PMID- 11045127 TI - [Lichen planus revealing a line of Blaschko]. PMID- 11045128 TI - [Level 2 analgesics for pain management: from the viewpoint of the neurologist]. PMID- 11045129 TI - [Dynamism of rheumatology: necessity and evidence]. PMID- 11045130 TI - [Progress in rheumatology]. PMID- 11045131 TI - [Augmentation of synthesized quantity of proteoglycans in vitro by articular chondrocytes treated with chondroitin sulfate]. PMID- 11045132 TI - [European multicenter study on effectiveness of chondroitin sulfate in gonarthrosis: a new look at biochemical and radiologic results]. PMID- 11045133 TI - [Beneficial effects of Chondrosulf 400 on pain and articular function in arthrosis: a meta-analysis]. PMID- 11045134 TI - [Metformin efficacious in poorly controlled diabetes mellitus type 2]. AB - Three patients, 1 man and 2 women, aged 60, 55 and 72 years, had an insufficient glucose regulation with insulin therapy or with sulfonylurea derivatives. They started metformin therapy, after which HbA1c decreased by 0.8-1.8%. One patient had to discontinue metformin therapy due to excessive diarrhoea. Many studies have shown the beneficial effect of metformin on glucose control. The recent UK Prospective Diabetes Study has proven the effectiveness of metformin for any diabetes related endpoint. Frequent side effects of metformin are nausea, abdominal discomfort and diarrhoea. Most side effects disappear after decreasing the dosage, although in 5% of patients diarrhoea only disappears after discontinuation of metformin. Lactic acidosis is a rare, serious adverse effect of metformin, which can be prevented by carefully observing the contra indications. PMID- 11045135 TI - [Current role of metformin in treatment of diabetes mellitus type 2]. AB - Metformin-associated lactic acidosis is not necessarily due to metformin accumulation. It appears that mortality in patients receiving metformin who develop lactic acidosis is mostly linked to underlying disease. It has been suggested that metformin should be the first-line agent for the treatment of obese type 2 diabetic patients since metformin was associated with a significant decrease in macrovascular events and a reduction of all-cause mortality in the United Kingdom Prospective Diabetes Study (UKPDS) in a substudy. However, in this substudy no significant decrease in microvascular complications was observed in obese subjects with intensive metformin therapy. In addition, the use of metformin in combination with sulfonylurea seemed to be associated with excess risk of diabetes-related and all-cause mortality in obese subjects. Due to the discrepant and contradictory nature of the results in the obese patients and a lack of power the UKPDS offered no decision for any drug for initial therapy of type 2 diabetes. The main message of the UKPDS is that lowering of the blood glucose to the normal range is beneficial irrespective of the hypoglycaemic agent used. A rational approach to therapy in a type 2 diabetes patient who fails to sufficiently lower blood sugar with diet and weight loss is to begin therapy with a sulfonylurea or metformin and to add another oral agent if the desired glycaemic control is not achieved. PMID- 11045136 TI - [Prevention of lactic acidosis due to metformin intoxication in contrast media nephropathy]. AB - Use of the oral antidiabetic drug metformin may cause lactic acidosis, a rare but life-threatening complication, especially in patients with renal function loss. Since intravenously administered iodide-containing contrast media may cause renal function disturbances precautions should be taken in metformin-treated patients for whom a radiological study with intravenous contrast media is considered. In diabetic patients who use metformin a serum creatinine concentration should be measured prior to the radiological study. If the serum creatinine is within normal limits (< 130 mumol/l), metformin can be continued and the examination performed. If the serum creatinine concentration is increased (> or = 130 mumol/l), metformin should be discontinued and replaced by another antidiabetic drug if necessary. The radiological procedure with intravenous contrast media should be postponed for 48 hours. If such a procedure cannot be postponed, additional measures to prevent lactic acidosis (hydration, monitoring of the renal function) should be taken. PMID- 11045137 TI - [Addition of beta blockers in chronic heart failure]. AB - Chronic heart failure is an increasing cause of hospital admission in the Netherlands and Belgium. Despite numerous medical treatment modalities, the mortality remains high. Recent placebo-controlled randomized studies suggest that the addition of beta-blockers in stabilized, optimally pretreated patients with chronic heart failure using angiotensin converting enzyme (ACE) inhibitors, diuretics and digitalis, is accompanied by an additional absolute decrease in mortality by about 5% and a relative decrease in mortality by about 35%. Also the number hospitalization frequency decreases. Initially, the beneficial effects of beta-blockers on symptoms are only minor or absent. During the initiation period some clinical deterioration may occur which has to be treated accordingly; these patients are, however, difficult to identify. Initiation has to be done using low doses and should be restricted to stabilized, optimally treated patients. Doses should only be increased every 2 to 4 weeks until target doses are reached. These findings must not be extrapolated automatically to all cases of heart failure, since patients in the trials may differ considerably from those encountered in general practice. PMID- 11045138 TI - [From gene to disease; from hemoglobin genes to thalassemia and sickle cell anemia]. AB - Haemoglobinopathies are the commonest autosomal recessive diseases in men. Mutations on the alpha and beta genes clusters, located on chromosome 16 and 11 respectively, have been strongly selected in many populations by the increased chance of survival of carriers in areas infested with malaria tropica. Unfortunately many of these mutations in homozygous or compound heterozygous forms generate severe phenotypes such as sickle cell disease and beta thalassaemia major. The population at risk for haemoglobinopathies is increasing in the industrialized areas of northern Europe. Without preventive measures a cumulative number of 1,000 severely affected patients can be expected in the Netherlands if information and carrier diagnostics are not efficiently offered at the GP level. A specialized laboratory for postnatal and prenatal diagnosis has been available in Leiden for more than 10 years now; however, couples at risk are only sporadically referred for counselling and/or prenatal diagnosis. PMID- 11045139 TI - [Diagnostic image (5) (Ileum pathology)]. AB - In a 28-year-old woman Crohn's disease was diagnosed. She had drug treatment but the situation deteriorated and ileocoecal resection was carried out. In the surgical specimen the diagnosis was confirmed. PMID- 11045140 TI - [Incidence of traumatic head or brain injuries in catchment area of Academic Hospital Maastricht in 1997]. AB - OBJECTIVE: To measure the incidence of traumatic head or brain injury in the catchment area of the Academic Hospital Maastricht (AZM), the Netherlands. DESIGN: Retrospective. METHOD: Data were collected about head injury patients who visited the emergency room of the AZM in 1997 by separate forms that were filled out for each patient who came to the emergency room. Data were added from admission records and radiology records. The AZM had a catchment area of approximately 231,000 people. RESULTS: The emergency room was attended by 1933 patients with traumatic head or brain injury. Head trauma without signs of brain injury was diagnosed in 1440 patients (74%) mild brain injury in 467 (24%) and moderate or severe brain injury in 26 (1%). The mean age was 30 years (range: 0 97) and 29% of all patients were below the age of 15. Two-thirds (67%) of patients were male. An X-ray of the skull was performed in 15% of the cases. In 7% of these X-rays a relevant abnormality was found. Eleven per cent of patients were admitted for observation. The incidence rate of traumatic head or brain injury in 1997 was 836/100,000 and the incidence of admission 88/100,000. The causes were a fall (43%), traffic accident (22%), violence (15%), sports injuries (7%), accidents during work (4%), or other/unknown (9%). CONCLUSION: Most patients with head or brain injury had mild injuries (99%, sole head injury or mild brain injury). Compared with other studies, the annual frequency of hospital admissions was low. PMID- 11045141 TI - [Assessment of day surgery in a district training hospital: safety, efficacy and patient's satisfaction]. AB - OBJECTIVE: To assess the quality of day surgery in the St. Antonius Hospital in Nieuwegein, the Netherlands. DESIGN: Prospective and descriptive. METHODS: During one year all patients treated by general surgeons in ambulatory surgery of the St. Antonius Hospital, Nieuwegein, the Netherlands (breast surgery (n = 232), hernia repair (n = 143), varicose vein surgery (n = 137), lymph node or lump excision (n = 85), (peri-)anal surgery (n = 70), ganglion surgery (n = 41), removal of bone implants (n = 41), laparoscopic cholecystectomy (n = 23), miscellaneous (n = 82); total 854) were evaluated by telephone questionnaires six weeks after surgery, to measure the following three aspects of quality of care: safety, efficacy and patient's satisfaction. Questions were asked about complications, visits to the emergency room, the outpatient clinic and the general practitioner and extra care at home. Unplanned clinical admissions following day surgery and re-admissions were registered. All outpatient clinic charts were also checked for complications. Whenever the registration of complications was incomplete the patient's general practitioner was contacted. All patients gave informed consent. RESULTS: After 854 planned day cases 823 patients (96.4%) returned home the same day. Reasons for clinical admission following day surgery were pain and/or nausea (n = 8), an operation late in the afternoon (n = 7), haemorrhage (n = 6), more extensive surgery than expected (n = 3), others (n = 7). Of all patients who returned home the same day and about whom the interview yielded adequate information (n = 656; 80%) 54 (7%) suffered from a complication (wound infection (n = 28), haemorrhage (n = 7), haematoma (n = 5), seroma (n = 3), phlebitis (n = 2), infection skin (n = 2), wound dehiscence (n = 2), others (n = 5)). Six patients were re-admitted. In the hospital and outpatient clinic 40 patients were seen without an appointment (6%) and 91 patients visited their general practitioner (14%). After surgery 84 (13%) patients were helped at home by friends or family. Of the group of patients who were successfully treated in day care 14% would have preferred an overnight stay. PMID- 11045142 TI - [Severe lactic acidosis due to metformin therapy in a patient with contra indications for metformin]. AB - A 52-year-old woman with a medical history of diabetes mellitus type 2, chronic alcoholism and liver function disorders was hospitalized because of complaints of haematemesis, abdominal complaints and dyspnoea. This was due to a severe lactic acidosis caused by acute alcohol intoxication and the use of metformin. With bicarbonate infusion and haemofiltration, the lactic acidosis disappeared, but she developed a distributive shock with multiple organ failure and died 23 days after admission. Lactic acidosis is a rare but serious adverse effect of metformin. Almost all patients described had contraindications to the drug, like renal failure, liver disease, alcohol abuse, and intercurrent conditions causing hypoxia or ischaemia. It is important to be aware of the circumstances in which metformin should not be prescribed. PMID- 11045143 TI - [Role of determination of QT dispersal intervals in anti-arrhythmia therapy]. AB - In recent years attempts intensified to identify and treat patients with an increased risk of sudden cardiac death caused by malignant ventricular arrhythmias. An elevated dispersion value of the QT interval of surface ECG suggests non-homogeneous repolarization and thus the presence of an arrhythmogenic substrate. In patients with ischaemic heart disease and confirmed malignant arrhythmias after onset of antiarrhythmic therapy changes in the dispersion of the QT interval occur, depending on the antiarrhythmic agent used and the effectiveness of treatment. In the authors group of 40 patients the value of dispersion of the QT interval declined significantly after introduction of effective antiarrhythmic treatment in patients treated with amiodarone and beta blocker. It may thus be stated that the value of the QT interval dispersion of surface ECG can be used not only as a non-invasive "marker" of sudden cardiac death but also as an auxiliary indicator of effectiveness of antiarrhythmic therapy. PMID- 11045144 TI - [Sonographic measurement of intimal thickness of the common carotid artery in diabetics]. AB - Incidence of atherosclerotic cardiovascular diseases in diabetics is known to be 2-4 times higher than in nondiabetic subjects. Intima-media-thickness of common carotid artery (IMT ACC) measured by B-mode sonography positively correlates with the severity of atherosclerotic changes in coronary arteries. This study compares sonographically assessed IMT ACC in patients with type 2 diabetes mellitus (27 patients, mean age 66.57 +/- 6.06 years) vs. 66 nondiabetic subjects (mean age 63.58 +/- 9.09 years). Intima-media thickness in diabetics was significantly higher when compared with the control group (0.826 +/- 0.29 mm vs. 0.647 +/- 0.26 mm). In the whole cohort of investigated subjects IMT ACC positively correlates with age. When adjusted for age, the mean IMT ACC level was significantly higher in men than in women. PMID- 11045145 TI - [Monitoring markers of bone turnover in multiple myeloma]. AB - PURPOSE: To assess the efficacy of bone turnover markers monitoring for prediction of overall survival and event-free survival in patients with multiple myeloma. To determine the value of initial measurement for differential diagnosis and evaluation of myeloma stage and activity. METHODS AND RESULTS: We evaluated 79 consecutive, previously untreated patients with myeloma and 24 with monoclonal gammopathy of undetermined significance. Urine excretion rates of pyridinoline (PYR) and deoxypyridinoline (DPYR) as markers of bone resorption and serum osteocalcin (OC) as marker of bone formation were measured at diagnosis and further repeatedly during the course of disease. High-performance liquid chromatography (HPLC) was used to measure urinary excretion rates of PYR and DPYR, serum OC levels was analysed using RIA method. Significant correlation was found between PYR and DPYR levels (p < 0.001) and between DPYR and OC (p < 0.05). Significantly higher urine PYR and DPYR (p < 0.001 and p < 0.01, respectively), but not serum OC, were found in myeloma comparing MGUS. In patients with MM PYR and DPYR levels were significantly higher in stage III vs. I and II (p < 0.01), the correlation with disease activity was not found. The grade of bone involvement according to RTG (osteolysis, osteoporosis, absence of bone lesions) was not reliable as a prognostic factor in our study. Univariate overall survival analysis showed prognostic significance for initial PYR (p < 0.05) but not for DPYR and/or OC. Rapid decrease of PYR (after 1 month of chemotherapy) was associated with the shortest median survival (608 days), however, significant difference was observed only comparing the patients in which PYR decreased 12 months after start of chemotherapy. In the univariate analysis increase of DPYR was the only variable significant for event-free survival (p < 0.05), increase of PYR tend to be significant (p < 0.1). CONCLUSIONS: We confirmed possible contribution of pyridinium cross-links for differential diagnosis of MM and MGUS and the correlation with advanced stage of MM. Initial measurement of PYR and monitoring of both PYR and DPYR during the course of myeloma could be helpful for prediction of overall survival and event-free survival. According to our experience OC is not useful for diagnosis, assessment of disease stage and activity and prediction of survival of patients with MM. PMID- 11045146 TI - [Clinical importance of determining levels of circulating transferrin receptors in blood]. AB - Circulating serum transferrin receptor level was measured using mouse monoclonal antibody against transferrin receptor (Orion Diagnostica, Finland) in 126 patients with various disorders of erythropoiesis and the results were compared to those obtained form control group consisted of 30 healthy volunteers with normal iron stores. Serum transferrin receptor level was significantly elevated in patients with iron deficiency and in all patients with hyperplastic erythropoiesis (hereditary spherocytosis, immune hemolytic anemia, beta thalassemia, myelodysplasia). Measurement of circulating serum transferrin receptor level was a sensitive indicator of iron depletion as well as a helpful parameter in differential diagnosis between iron deficiency and anemia of chronic disease where circulating transferrin receptor level was not elevated. Index transferrin receptor/ferritin calculated as a ratio of circulating serum transferrin receptor level to log serum ferritin level was a more sensitive parameter than measurement of serum transferrin receptor not only for determination of patients with anemia of chronic disease, but also for discrimination of patients with elevated serum transferrin receptor level due to true iron deficiency from those with high serum transferrin receptor level caused by relative iron deficiency in hyperplastic erythropoiesis. PMID- 11045147 TI - [Vitamin D deficiency in hospitalized patients]. AB - Vitamin D deficiency is one of the important risk factors for the development of osteoporosis and fractures. The high prevalence of hypovitaminosis in elderly people in old age pnesioners homes was proved in several investigations, similarly as the favourable effect of vitamin D (800 IU/day) and calcium supplementation on a decline of fracture risk. Risk factors of hypovitaminosis such as an inadequately varied diet; low exposure to sunlight, chronic liver and kidney disease and treatment affecting the metabolism and clearance of vitamin D are very frequent in elderly patients hospitalized in medical departments. In the submitted trial the authors assessed in a group of 38 patients, mean age 70 years, hospitalized at the medical department at the end of the winter period the vitamin D3 serum level. They found a significant reduction of the concentration of 1.25 hydroxyvitamin D3 (p < 0.01) in the investigated group. The results of the trial, along with data in the literature on the high prevalence of hypovitaminosis D in the European population, indicate the need to introduce this simple cheap and safe therapeutic modality into routine practice. PMID- 11045148 TI - [Salmon calcitonin in the treatment of osteoporosis]. AB - Salmon calcitonin is frequently used in the treatment of osteoporosis. Via specific receptors on osteoclasts it reduces bone absorption and has also an osteogenic effect. Treatment with calcitonin leads to an increase in bone density, drop of markers of bone turnover and reduction of risk fractures. In the submitted trial the authors investigated the influence of nasal calcitonin administered in amounts of 100 IU in three-month intervals, on bone density. The group comprised 59 patients, mean age 64 years. The mean period of treatment was 2.5 years. Treatment led to a significant increase of bone density in the region of the lumbar spine and neck of the femur. Treatment was well tolerated and the authors did not record new manifest fractures or side-effects of treatment. PMID- 11045149 TI - [Why evaluate the quality of life in patients with bronchial obstruction?]. AB - Quality of life, more precisely health related quality of life, is gaining basic importance in evaluating of patient's health status and results of medical interventions. On the example of patients with bronchial obstruction is demonstrated that quality of life evaluation, although not usually used in clinical practice yet, is in significant correlation with some of usual clinical measures; moreover, it brings otherwise inaccessible information about patient's subjective assessment of his disease and his health situation. PMID- 11045150 TI - [The little known Fay syndrome (benign carotidynia)]. AB - The authors describe the little known Fay syndrome (benign carotidynia, extracranial vascular cephalea). They emphasize the importance of a careful case history (in particular sore throat--usually unilateral), with careful physical examination (palpation sensitivity of the carotid artery beneath the bifurcation). The authors assume that the syndrome is not as rare but that it is rarely correctly diagnosed. The drug of choice are non-steroid antirheumatic drugs. The authors describe in an epicritical manner eight of their own observations made during the last three years. PMID- 11045151 TI - [Leukemic transformation of polycythemia vera after treatment with hydroxyurea and chromosome 17 abnormalities]. AB - The authors present two patients with polycythemia vera where they recorded after several years' treatment with hydroxyurea development of acute myeloblastic leukaemia. In both instances they found, associated with leukaemia, abnormalities of chromosome no.17, in one case meeting criteria of the so-called 17p-syndrome. Progression of polycythemia vera into acute leukaemia is explained by the possible association with the long-term use of the drug and loss of chromosomal material (short arm of chromosome 17), the part where genes important in the process of leukaemogenesis are located. The authors draw attention to contemplated long-term administration of hydroxyurea to young patients with polycythemia vera. As cytogenetic analysis is a suitable method for evidence of progressing polycythemia vera into acute leukaemia, dynamic follow up of chromosomal changes is necessary, in particular in patients where long-term treatment with hydroxyurea is assumed. PMID- 11045152 TI - [Drug interactions in coumarin anticoagulation]. AB - The clinically most important negative properties of coumarin anticoagulants are their numerous interactions with other drugs. Data in the literature on these interactions are frequently controversial. The causes of this phenomenon are a) the complicated path from ingestion of anticoagulants to their clinical effect and laboratory results, b) interindividual differences in enzyme activity participating in vitamin K metabolism, in the synthesis of hemocoagulation factors and in the biotransformation of coumarin derivatives, c) the different clinical state of the patient on the one hand and the healthy volunteer on the other, d) differences in the biotransformation of different coumarin derivatives and their optic isomers. In the submitted paper the authors describe pharmacokinetic and pharmacodynamic mechanisms of drug interactions of coumarin anticoagulants and their importance for clinical practice. In the conclusion recommendations are submitted to reduce the risk of these interactions. PMID- 11045153 TI - [Population screening for colorectal carcinoma by examining feces for occult hemorrhage]. AB - Colorectal carcinoma is in the western world one of the most frequent deaths from malignant disease. A reliable way how to reduce the mortality from this disease is early diagnosis by screening in the population, the most suitable method being examination of the faeces for occult bleeding. During the last five years in the world literature results of several long-term and well elaborated studies were published focused on screening of colorectal carcinoma by detection of occult haemorrhage in the faeces. There were three American and four European papers from which ensued: 1. In the examined population in the mentioned investigations a reduction of the mortality from colorectal carcinoma was recorded as compared with a control group. 2. In the investigated population also a reduced incidence of advanced colorectal carcinoma was observed (Dukes D). 3. The optimal ratio of sensitivity and specificity was obtained by the HemeSelect test (immunodiagnosis of human haemoglobin) and the combined test (Hemoccult II Sensa + HemeSelect). Large scale examination by these tests was not verified in major population surveys so far. 4. Increasing of the sensitivity of tests using guaiacum resin by rehydration leads to a drop of the specificity of the examination and an enormous increase of coloscopic examinations and the cost of screening. 5. Part of the population with a high risk of colorectal carcinoma should be subjected to large scale examination. The problem remains its correct selection. 6. The adherence of patients to the examination differed in different trials. The increasing number and pretentiousness (dietary restrictions) of the examination reduces proportionately the percentage of cooperating subjects. Examination of the faeces for occult haemorrhage thus can be recommended for national programmes for screening of colorectal cancer. PMID- 11045154 TI - [Assessment of risk for the development of manifest atherosclerosis and indications for hypolipidemic therapy (global risk assessment)]. AB - The estimated risk of development of complications of atherosclerosis (coronary or cerebral ischaemia) is one of the activities every physician should include in the diagnostic and therapeutic algorithm. Treatment with hypolipidaemic agents is indicated according to the European consensus if the danger of a coronary or cerebral event is 20%/10 years. The objective of preventive provisions should be reduction of this risk to < 5%/10 years in younger subjects (< 45 years in men and < 50 years in women) and < 10%/10 years in elderly subjects. These recommendations obviously do not apply to secondary prevention (in case of complications of atherosclerosis) where treatment with hypolipidaemic agents is indicated already when the LDL-cholesterol level is 2.3 mmol/l or when triacylglycerols are 2.3 mmol/l, combined with a drop of HDL-cholesterol to < 0.9 mmol/l. PMID- 11045156 TI - [Is the existence of integrated internists necessary and why?]. PMID- 11045155 TI - [The first 100 liver transplantations in Brno]. PMID- 11045157 TI - [Changes in left ventricular function during chemotherapy with doxorubicin]. AB - The authors investigated by echocardiography changes of left ventricular function in 79 patients with non-Hodgkin lymphoma or Hodgkin's disease who were treated by chemotherapy containing doxorubicin. In 22% patients they diagnosed during treatment a significant gradual decline of the left ventricular ejection fraction (change > 10% or a drop of EF below 50%) after a cumulative dose of 185 +/- 52 mg/m2 doxorubicin (median 200 mg/m2). Changes of the ejection fraction were at the expense of an increasing endsystolic left ventricular volume. These changes correlated closely with the increase of endsystolic stress of the left ventricular wall (r = -0.87). After administration of a cumulative doxorubicin dose of 100 mg/m2 a significant deterioration of indicators of diastolic filling of the left ventricle occurred--of the isovolumic relaxation period (IRP) and deceleration time (DT). The contribution of these early changes of diastolic function of the left ventricle for assessment of the risk of development of systolic dysfunction is not unequivocal. Prolongation of IRP > 12% as compared with the baseline value and at the same time above 95 ms had a 50% sensitivity, 50% specificity, a 26% positive prediction value a 74% negative prediction value and 50% diagnostic accuracy, change of DT > 13%, and at the same time prolongation above 200 ms had a 47% sensitivity 55% specificity, a 22% positive prediction value, a 78% negative prediction value and a 53% diagnostic accuracy. The authors found a significant relationship between the assessed changes of left ventricular function and clinical and haemodynamic indicators. Only the patients' age correlated significantly with IRP and DT values. Changes of the left ventricular ejection fraction were not associated with clinical signs of cardiac failure, they did not call for cardiological intervention and did not affect the course of chemotherapy. Contrary to some statements in the literature, the authors did not consider the described functional changes as sufficient to justify interference with chemotherapy. On the other hand, their importance for the development of late sequelae of chemotherapy with anthracyclines in adult patients must be based on long-term prospective follow-up. PMID- 11045158 TI - [The Ewing test for autonomic neuropathy and spectral analysis of heartrate variability aid in the diagnosis of Charcot's osteoarthropathy]. AB - The objective of the work was to evaluate the contribution of examining autonomic neuropathy in diabetic patients to early diagnosis of Charcot's osteoarthropathy by classical Ewing's tests, as well as the more recent method--spectral analysis of heart rate variability. The authors examined 18 diabetic patients in the early stage of Charcot's osteoarthropathy and the results were compared with a group of 30 subjects matched for age and sex. The results of examination by Ewing's test (heart rate variability during deep breathing, in orthostasis and during Valsalva's manoeuvre and blood pressure changes during orthostasis) revealed autonomic neuropathy in all examined patients, in one subject incipient neuropathy and in 17 of 18 patients manifest or severe neuropathy. The patients differed from controls highly significantly in all parameters of Ewing's tests with the exception of parameter 30:15 in orthostasis. The greatest sensitivity was recorded in the examination of the I-E difference during deep respiration. RRmax/RRmin and the brake index in orthostasis and Valsalva's ratio. The lowest sensitivity was recorded in the examination of the orthostatic fall of blood pressure. On spectral analysis the patients differed highly significantly from controls in all investigated parameters, the highest discriminating value was found in parameters of the total spectral power in the standing position (2) and the power in the low-frequency area in position 2, the first parameter alone was correctly discriminated in 94%. SUMMARY: Examination of autonomic neuropathy significantly improves the diagnosis of Charcot's osteoarthropathy. In addition to the classical Ewing tests spectral analysis of heart rate variability proved also a suitable method for its evaluation. PMID- 11045159 TI - [Treatment with interferon and autoimmunity]. AB - Interferons are commonly used, frequently already as a standard drug, in a relatively wide spectrum of indications in tumourous and non-tumourous diseases. The numerous undesirable therapeutic effects include also relatively frequent induction of autoimmune processes which, however, only rarely is associated with marked clinical manifestations. The author presents an account on two patients from their own clinical practice treated with interferon alpha who developed complications which in the author's opinion may be associated with autoimmunity (a patient with thrombocytopenia responding well to immunosuppressive treatment, originally treated on a chi ount of renal carcinoma, and a female patient with pulmonary sarcoidosis treated for essential thrombocythemia). The authors present also a brief review of the literature on autoimmunity induced by interferon. PMID- 11045160 TI - [A patient with Madelung's disease at the internal medicine department]. AB - Madelung disease, benign symmetrical lipomatosis, is a rare disorder of unknown etiology. It is characterized by the accumulation of fat deposits around the neck and upper body. It is often associated with glucose intolerance, liver disease, and malignant tumors of the lungs. This disease normally affects middle-aged males with a history of alcohol abuse. Surgical removal of the lipomatose mass is the treatment of choice. A case of Madelung disease in a 53-year old man with hepatopathy and respiratory failure is presented. PMID- 11045161 TI - [Fulminant liver failure as a sequelae of acute dystrophy of the right hepatic lobe]. AB - A 38-year-old female patient developed signs of fulminant hepatic failure, caused by extensive, spontaneous, devastating ischaemia of the right liver lobe. The patient survived the acute attack. During the subsequent eight years hypertrophy of the intact left liver lobe occurred which eventually completely replaced the loss of the dominant right lobe. PMID- 11045162 TI - [Hypertrophy of the left ventricle--etiopathogenesis, clinical consequences and prognosis]. AB - The left ventricular hypertrophy is a risk marker of the cardiovascular morbidity and mortality in hypertensive patients--it contributes to sudden death, myocardial infarction, myocardial ischemia, heart failure, arrhythmias, left ventricular diastolic dysfunction, stroke and renal failure. The mechanisms by which the heart hypertrophy increases the risk of cardiovascular morbidity and mortality, however, is not completely clear yet. Pressure overload (resulting in the concentric hypertrophy) and volume overload (resulting in the eccentric hypertrophy) of the left ventricle play a significant role in the development of the hypertrophy of the left ventricle. Other risk factors, stimulating left ventricular hypertrophy, include growth factors, genetic predisposition, age, obesity, hyperinsulinemia and anemia. The hypertrophy of left ventricle most often occurs with hypertension, cardiomyopathy and aortic stenosis. Several clinical studies evaluated functional consequences of the reduction of the ventricular hypertrophy and found out that the function of the left ventricle to be improved in hypertensive patients who had undergone an effective and long-term antihypertensive treatment. However, these studies did not differentiate whether for the improvement in the function of left ventricle was the matter of the reduction of the left ventricular mass or whether it was because of the decrease of the arterial pressure during the period of anti-hypertensive treatment. On the basis of the literature studied we can emphasize that the reduction of myocardial hypertrophy resulting from a specific antihypertensive treatment appears to be more favourable than harmful for the heart's pump performance. PMID- 11045163 TI - [Non-invasive detection of failure of thrombolysis in acute myocardial infarct]. AB - Despite advances in thrombolytic treatment of acute infarction in a major proportion of patients occasionally despite its administration reperfusion of the heart muscle does not occur which implies a negative impact on the long-term prognosis of these patients. With regard to the fact that invasive diagnostic methods are not available in the field, the basis of non-invasive diagnosis and stratification of patients for further reperfusion treatment is assessment of the symptomatology (chest pain) and frequent controls of 12-lead ECG, although controlled trials evaluating their validity for decisions in acute situations are not quite unequivocal. With the development of quantitative bedside enzyme diagnosis it may be anticipated that it will play a more important part in the differentiation of reperfusion failures. The basis of diagnosis in thrombolysis failure is in contemporary clinical practice simultaneous evaluation of all available non-invasive indicators. PMID- 11045164 TI - [Vasoneuroses]. AB - The term vasoneuroses comprises functional disorders of peripheral vessels, in particular arterioles, manifested by vasospasms on the periphery of the upper and lower extremities, more rarely the nose and ears. The main representant of these diseases is a disorder manifested by episodic attacks of ischaemia of the fingers, in particular of the upper extremities--Raynaud's phenomenon (RP) which has two forms: primary RP (Raynaud's disease) where no other basic disease is diagnoses during a two-year period. secondary RP (Raynaud's phenomenon) as an associated symptom of other, in particular systemic diseases. The etiology and pathology of Raynaud's phenomenon has not been elucidated satisfactorily so far. With regard to the variety and scope of detected functional and morphological abnormalities RP is rather multifactorial, caused by an unbalanced action of local and systemic factors affecting the sensitivity of the vascular wall to spastic stimuli. Treatment therefore remains symptomatic. It is restricted to administration of vasodilatating agents, in more severe cases sympathectomy is considered; it is important to rule out another basic disease. Vasoneuroses include also acrocyanosis, livedo reticularis and erythromelalgia. In the case of erythromelalgia it is important to rule out secondary causes (hypertension and polycythemia vera), otherwise it is not necessary to use pharmacological means to influence these diseases because of their relatively harmless course. PMID- 11045165 TI - [Present status of fibrates in the treatment of hyperlipoproteinemias]. AB - Fibrates are hypolipaemic agents with a broad spectrum of action and long-term safety. They reduce the cardiovascular mortality and morbidity and can induce also regression of coronary atherosclerosis. They lower the plasma concentration of total and LDL-cholesterol, triacylglycerols, and produce a rise of HDL cholesterol. Moreover they can reduce the level of fibrinogen, lipoprotein/a/, PAI-1, uric acid and the concentration of highly atherogenic small dense LDL3. Their effect is mediated by activation of "peroxisome proliferator-activated receptors" (PPARs) in the cell nucleus. At present we find on the market second generation fibrates as well as modern micronized medicaments of the third generation. They are indicated for treatment of patients with hypertriglyceridaemia, combined hyperlipidaemia and diabetic dyslipidemia. PMID- 11045166 TI - [Micronized fenofibrate and LDL-cholesterol subfractions]. AB - LDL-cholesterol subfractions have a different atherogenity; the most atherogenic are small LDL3 called small dense LDL. A clear relationship was proved between their concentration and early manifestation of ischaemic heart disease. In some instances they were found to act as an independent risk factor of cardiovascular disease. Their concentration depends to a great extent on the triacyglycerol concentration. They are found most frequently in patients with combined hyperlipidaemia, in associated metabolic syndrome and in patients with type 2 diabetes mellitus. Their plasma concentration can be reduced by non pharmacological methods (restriction of animal fats, reduction of body weight, physical activity) as well as by pharmacological means (in particular fibrates). Micronized phenofibrate reduces significantly the concentration of small LDL3 and thus contributes to normalization of dyslipoproteinaemia and reduction of the risk of cardiovascular complications. PMID- 11045168 TI - [Importance of determining the propidium-iodide index of plasmacytes in multiple myeloma. II. Relation to disease extent and activity]. AB - The authors evaluated in a group of 50 patients with multiple myeloma (MM), examined when the diagnosis was established before onset of treatment, and in a group of 85 patients examined in different stages of MM the relationship of proliferative characteristics of myeloma plasmocytes assessed by means of the propidium-iodide index PI/CD38, PI/"UHKT" and PI/B-B4(CD138) with the progression, "performance status" and the activity of the disease. With the exception of a different value of the PI/CD38 index between stages 1 and 3 evaluated according to Durie-Salmon, in the whole group of patients no relationship was found with the progress of the disease. When evaluating the whole group the authors observed a difference between sub-stages A and B (i.e. between patients without and with a serious impairment of renal function) when using indexes PI/CD38 and PI/B-B4(CD138). Only in the whole group of 85 patients the authors found a significant relationship of all proliferation indexes (PI/CD38PI/"UHKT" and PI/B-B4) with the "performance status" according to ECOG score to or > or = 3 vs. < 3. In both groups there was a very close statistically significant relationship without the activity of the disease whichever of the three proliferation indexes was used. It was revealed that patients with the active but stable form of the disease have different PI/CD38 and PI/B-B4(CD138) indexes within the framework of the same stage of MM (stages 1 + 2 vs. 3). From the investigation ensues that examination of the PI proliferation index, in particular PI/B-B4(CD138) or possibly PI/CD38 using multiparametric flow cytometry is a valuable method extending hitherto used examination procedures in MM, and that it replaces adequately former autoradiographic and microscopic immunofluorescent techniques. PMID- 11045167 TI - [Importance of determining the propidium-iodide index of plasmacytes in multiple myeloma. I. Relation to selected laboratory indicators of the disease]. AB - In a group of 85 patients with multiple myeloma (MM) incl. 50 patients examined at the time when the diagnosis was established the relationship of the values of the propidium-iodide index (PI index) of myeloma plasmocytes of the patients were examined by flow cytometry using the double staining method, and selected laboratory parameters of the disease were analyzed. It was revealed that the values of the PI index examined with the aid of different "identification" monoclonal antibodies did not differ significantly. The median and average value of the PI index was in the whole group for PI/CD38 2.3 and 2.3%, for PI "UHKT" 1.8 and 1.8% and for PI/B-B4(CD138) 2.2 and 2.4%. In the group of patients examined at the time when the diagnosis of MM was established before chemotherapy the median and average value of the PI/CD38 index was 2.0 and 2.2%, PI/"UHKT" was 1.5 and 1.6%, PI/B-B4 (CD138) 2.6 and 2.5%. In the two analyzed groups no statistically significant correlations of PI/CD38,/"UHKT" and B-B4(CD138) index were found with the number of granulocytes, thrombocytes, immunochemical type and the value of the serum M-component, and levels of S-beta 2 microglobulin, S-urea, S-creatinine, S-calcium, and S-lactic dehydrogenase. A significant positive correlation was found in both groups with the level of S-thymidine kinase, in the whole group of indexes PI/CD38 and PI/B-B4(CD138) with the severity of anaemia, index PI/CD38 correlated with S-albumin and index PI/B-B4(CD138) with the percentage ratio of plasmocytes in bone marrow. It was revealed that examination of the propidium-iodide index is a suitable and prompt method for evaluation of proliferative properties of the myeloma population. PMID- 11045169 TI - [Poor response in patients with de novo acute myeloid leukemia and erythroblastic or megakaryocytic dysplasia to treatment with standard doses of cytosine arabinoside and daunorubicin]. AB - Erythroid and/or megakaryocytic dysplasia (EMD) was evaluated in diagnostic bone marrow smears of 37 consecutively treated cases with de novo acute myeloid leukemia (AML) M0-M2 and M4-M5 types and three newly diagnosed cases of refractory anaemia with excess of blasts in transformation. This evaluation was possible in 38 of 40 (95%) patients and 17 cases were categorized without EMD and 21 cases with EMD. One or two cycles with 3-4 doses of daunorubicin 45 mg/m2/d and cytosine arabinoside 200 mg/m2/12 h x 14 lead to complete remission in 13 of 16 cases without EMD but only in 4 of 16 cases with EMD (p = 0.004). However, 10 of 13 patients with EMD reached complete remission with 2000 mg/m2/12 h x 10 of cytosine arabinoside plus daunorubicin. After intensive consolidations 20 patients under 65 years with EMD showed significantly worse overall survival (p = 0.017) with a median 11.5 months, while the median survival was estimated 66.8 months in 15 cases under 65 years without EMD. The proposed categorization of de novo AML patients according to the EMD seems to be useful for selection of optimal induction therapy, clinically relevant for survival and might reflect two biologic groups of AML arising in two different stages of differentiation. PMID- 11045170 TI - [Personal experience with determination of carbohydrate-deficient transferrin]. AB - Assessment of carbohydrate-deficient serum transferrin CDT) is considered a very useful indicator of alcohol abuse. There is a number of methods for assessment of CDT. In clinical practice most frequently assessment of the percentage ratio of CDT in transferrin is used. In our hospital we assessed CDT by the method of homogeneous immunoanalysis (Boehringer Mannheim (CDT-BM). Because we obtained a relatively high percentage of false results and because we found in the literature reference to a new method of homogeneous immunoanalysis (The Sanqui BioTech-CD-SB) with a different cut-off, we decided to compare the results of the estimations by the two methods and assess the percentage of false results. We examined a group of 49 patients incl. 16 who admitted alcohol abuse (> 60 g alcohol more than four times per week). As anticipated, we found that the %CDT assessed by the CDT-BM method was significantly higher than the percentage CDT SB. After classification of the group into sub-groups with regard to alcohol intake the two sub-groups differed significantly only in values of CDT-SB and CDT BM. In the group of patients with alcohol abuse we found relations between CDT-MB and indicators of hepatic lesions. In CDT-SB we found only an association with AST. This finding could suggest a greater specificity of CDT-SB. We confirmed data in the literature that GMT is independent on CDT and the mean corpuscular volume is independent on CDT in subjects with alcohol abuse. In abstainers who negated alcohol intake we found also when assessing CDT-SB a significant gender differentiation which is described in the literature (the reason is probably the fact that the CDT-SB method analyzes, contrary to CDT-BM, only asialo,-monosialo and disialylic isoforms of transferrin and women have higher levels of monosialylic forms). In our group the examination of %CDT by the new method, the Sanqui Biotech for alcohol abuse, had an almost absolute specificity and sensitivity. Contrary to the older Boehringer-Mannheim method we did not record any false increase in any patient with signs of hepatopathy nor any false negative results). We assume that the described methodical innovation of the analysis could facilitate the differential diagnosis of various diseases in different medical disciplines (internal medicine, neurology, psychiatry, assessment of work capacity). PMID- 11045171 TI - [Antioxidants and cardiovascular diseases in the Czech population]. AB - The authors studied the possible association between plasma antioxidants and the high rates of cardiovascular diseases in the Czech Republic. The report has three parts. First, plasma antioxidants levels were compared in a random sample of population of two Czech districts (70 men and 66 women) and in British civil servants (246 men and I 12 women). Second, plasma antioxidants were compared in random samples of men aged 45-64 in Pardubice (Czech Republic, n = 185) and Augsburg (Bavaria, n = 142). Both Breat Britain and Bavaria have substantially lower mortality from cardiovascular disease than the Czech Republic. Third, a case control study examined the relation between low levels of plasma antioxidants and the risk of non-fatal myocardial infarction (52 cases and 144 controls, all males). Blood samples were taken, stored and analysed under strictly standardised conditions. Geometric means of beta-carotene, alpha tocopherol and the alpha-tocopherol/total cholesterol ratio were substantially lower in Czechs than in Brits (all p-values less than 0.001). For example, the mean concentration of beta carotene in plasma of Czech men was only a half of that in British men (0.39 vs. 0.77 mumol/l). Similarly, mean alpha- and beta carotene and lycopene among Czech men were substantially lower than in Bavarian men. Mean homocysteine was higher in Czechs (10.5 mumol/l) and in Bavarians (8.9 mumol/l, p < 0.001). Means of vitamin E were similar. In the case-control study, the risk of myocardial infarction was elevated among men with below median plasma levels of beta carotene (age adjusted odds ratio 3.33, 95% confidence interval 1.43-8.33) and alfa-tocopherol (odds ratio 1.89, 95% confidence interval 0.94 3.85). The low levels of carotenoids and high levels of homocysteine in plasma of Czechs may reflect low dietary intakes of fresh fruits and vegetables. Antioxidants or factors related to their dietary intakes may be one of the causes of the high cardiovascular mortality in the Czech Republic. PMID- 11045172 TI - [Pain and rheumatoid arthritis]. AB - OBJECTIVE: Chronic pain is an important part of the clinical picture in patients with RA and therefore the authors attempted to assess the relationship of pain of patients with RA to the inflammatory process, articular damage and to predict the variance of pain in relation to psychosocial, demographic factors and functional disability. METHOD: Pain was examined using the Nottingham Health Profile (NHP), psychological health by means of the General Health Questionnaire (GHQ) and disability was assessed by the Health Assessment Questionnaire (HAQ). RESULTS: The authors examined 160 patients with RA with a mean age of 48.34 +/- 12.13 years and persistence of the disease for 21.32 +/- 15.61 months. Pain was significantly related to clinical manifestations of inflammation and articular damage. The relationship between pain and laboratory manifestations of inflammation (CRP, FW) was not very close. The relationship between age and pain and between persistence of the disease and pain was not statistically significant. Pain correlated significantly with functional disability (HAQ) (r = 0.54) and the articular index (RAI) (r = 0.58). Pain did not correlate with manifestations of depression (r = -0.15), but correlated significantly with anxiety (r = -0.35) and social dysfunction (r = -0.36). Among the investigated parameters (demographic factors, GHQ and HAQ) the most marked variation of pain was caused by HAQ (32%), GHQ (8%). Almost unrelated were demographic parameters (0.6%) (age, persistence of disease, education lifestyle, and gender). PMID- 11045173 TI - [Anticoagulation therapy and bone metabolism]. AB - The effect of anticoagulation treatment with coumarin and heparin on bone metabolism is in recent years frequently discussed in professional journals. Its ever expanding use in the treatment of patients deserves therefore attention in particular in patients having prolonged anticoagulation treatment. In the submitted paper the authors investigated the effect of Warfarin on some indicators of bone metabolism in a group of 17 patients. In the investigated parameters such as estimation of parathormone, serum calcium, phosphatemia, isoenzyme of alkaline phosphatase they did not reveal any statistically significant changes during coumarin treatment. PMID- 11045174 TI - [Indicators of oxidative stress in cardiovascular diseases. I. Lipoperoxidation]. AB - The level od malondialdehyde (MDA), thiobarbituric acid reactive substances (TBARS) respectively, in blood is changing very quickly and therefore isoprostanes scam to be better markers of atherogenesis. Oxidized LDL belong to the most important reasons of the origin of foam cells and therefore of atherogenesis. That is why the inhibition of the oxidation of LDL particles should belong to the standard therapy of cardiovascular diseases. The value of antibodies against oxidized LDL is difficult to interpret, because they are multifactor dependent, especially under genetic control. Antibodies against LDL modified by malondialdehyde (MDA-LDL) are increased in smokers and individuals with a rapid atherosclerotic progression. HDL-cholesterol protects LDL through the oxidized lipid exchange. Oxidized HDL formed during this process has then an increased clearance from circulation. PMID- 11045175 TI - [Indicators of oxidative stress in cardiovascular diseases. II. Nitrogenous substances, AGE-substances and antioxidants]. AB - By oxidation of nitrogen oxide nitrites and nitrates are formed which can be considered markers of inflammation. AGE-substances (products of advanced glycation) are formed not only from proteins (e.g. Hb-AGE) but also from lipids (AGE-LDL) and DNA (AGE-DNA) and lead to an increased number of mutations. They are toxic and are deposited in the vascular wall. They cause coagulopathy, stimulate cytokines and inactivate nitrogen oxide and thus prevent relaxation of the vascular wall. Extracellular superoxide dismutase (EC-SOD) has a variant (mutation Arg213Gly with a 4% incidence) which causes deterioration of the cardiac prognosis. Selenium deficiency leads also to an increased frequency of cardiovascular diseases. PMID- 11045176 TI - [Pathology of the spleen in ultrasonographic imaging]. AB - Authors provide review on diffuse and focal splenic lesions, traumatic injuries, contribution of ultrasonography in staging of malignant lymphomas and ultrasound guided percutaneous biopsy. Special attention is given to the question of normal spleen size and diagnosis of splenomegaly. In 230 patients (98 males) aged 18-72 years (median 34.2) witch underwent complex preventive examination, maximal longitudinal diameter (LDmax) of the spleen were measured. Using linear regression analysis relation of spleen size and body high, weigh and body mass index (BMI) was evaluated. Median LDmax was 10.1 cm, range 7.8-13.5 cm. As a reference limit authors accept 95% confidence interval of median: 7.9-12.3 cm. Statistically significant correlation (p < 0.01, r = 0.31) between LDmax and body high was found out. Ultrasonography because of diagnostic efficiency is suggested the method of choice in diagnosing pathologic involvement of the spleen. PMID- 11045178 TI - [Role of physiopathology in medical aspects of large clinical studies]. PMID- 11045177 TI - [Hemophilic arthropathy as a multidisciplinary problem]. AB - The authors have at present on their records in Slovakia 422 patients with haemophilia type A, i.e. 7.9 per 100,000 population, 76 cases of haemophilia type B, i.e. an incidence of 1.4, 189 patients with von Willebrand's disease with an incidence of 3.6 and 215 patients with hypoconvertinaemia with an incidence of 3.98 per 100,000 population. The authors present an account on the most frequent complications of haemophilia with special attention to haemorrhage into the joints and the development of serious haemophilic arthropathies. They emphasize the great contribution of team work of different medical specialists who participate in the diagnosis and treatment of these diseases. PMID- 11045179 TI - [Serum leptin levels in female patients with protein-calorie malnutrition and its relation to biochemical indicators of nutritional status]. AB - Leptin is a protein hormone produced by adipocytes. Its serum concentrations in the most of cases positively correlate with total body fat content. Serum leptin levels are increased in obese in comparison with lean subjects. Leptin levels in females are two or three times higher than in body mass index and age-matched males. The aim of our study was to investigate the serum leptin concentrations in females with protein-caloric malnutrition of various aetiology, the relationship between leptin levels and biochemical nutritional parameters and to compare the values with those of age-matched control group. Totally 12 patients with malnutrition of various aetiology and 14 control age-matched healthy subjects were included into the study. It was found that serum leptin levels were significantly decreased in malnutrition group in comparison with control group (3.09 +/- 1.33 ng.ml-1 vs 9.42 +/- 2.76 ng.,l-1, p < 0.05). A significant decrease in body mass index, serum total protein, albumin, prealbumin, cholesterol and triglycerides concentration was also found in malnutrition in comparison with control group. While a strong positive correlation between leptin concentrations and body mass index was found in control group (r = 0.72, p < 0.05), no statistically significant relation between leptin and body mass index was observed in malnutrition group. Furthermore, no significant relationship was found between serum leptin levels and serum total protein, albumin, prealbumin, cholesterol and triglycerides neither in control or in malnutrition group. We conclude that serum leptin levels in patients with malnutrition are significantly decreased in comparison with age-matched healthy control subjects. The loss of positive correlation between leptin and body mass index in malnutrition group is explainable by the changes of body composition in patient with malnutrition. The body mass index value in these patients reflects the total body fat content less precisely than in healthy controls. PMID- 11045180 TI - [Leptinemia in individuals with hypertension (pilot study)]. AB - During the last decade several papers were published where obesity in included among the building stones of the so-called metabolic cardiovascular syndrome (along with hypertension, dyslipidaemia, impaired glucose tolerance and hyperinsulinaemia). Several months ago it was also revealed that some patients with the metabolic syndrome suffer from hyperleptinaemia. Leptinaemia is considered by some authors as independent indicator of the risk of accelerated atherosclerosis. The cause of these hypothesis were (in addition to the known conclusions on the occurrence frequent incidence of leptin resistance and insulin resistance) in particular the results of experimental studies where evidence was provided that infusion induced hyperleptinaemia leads in animals to hypertension due to its direct effect on sympathicotonia and peripheral vascular resistance. The authors of these hypothesis assume that hyperleptinaemia (in particular in subjects with metabolic syndrome) in one of the basic causes of hypertension which is frequently encountered in these patients. OBJECTIVE: To assess the relations between leptinaemia and the blood pressure reading (actual and mean values) and leptinaemia and the classification of hypertension according to WHO. METHOD: The authors examined 35 hypertensive subject and 10 subject with tetanies (without hypertension). The blood pressure was assessed under standard conditions (rest, semi-recumbent position, three readings). The mean blood pressure readings during the last three months were obtained from case records. Leptin was assessed by the ELISA method of Bio Vendor Co. RESULTS: The group of hypertonic can be classified as subjects with mild obesity (BMI 30.1). The values of leptin were elevated but did not differ significantly from those of the normal population. No correlations were found between leptinaemia (incl. values calculated for BMI) and the actual and mean blood pressure readings. No correlations were found between leptinaemia (incl. BP calculated with regard to BMI) and the stage of hypertension according to WHO. CONCLUSION: Hypertonic subject do not differ significantly as to the serum leptin concentration from the general population. Leptinaemia does not correlate with the actual or mean blood pressure reading nor with stage of hypertension according to the WHO classification. Thus the authors did not confirm the hypothesis on the fundamental effect of leptinaemia on the genesis and development of hypertension. It is probable that leptin is only one of the many factors which have an impact on blood pressure. PMID- 11045181 TI - [Long-term treatment of combined hyperlipidemia with a combination of fluvastatin and fenofibrate]. AB - The long-term efficacy and safety of fluvastatin monotherapy was compared with that of the combination of fluvastatin and fenofibrate in 104 patients with coronary heart disease and combined hyperlipidemia in an open, randomised, parallel group, clinical study of 78 weeks duration. Combined hyperlipidemia was defined as LDL-cholesterol 4.1 mmol/l and higher and triglycerides between 2.5 and 4.5 mmol/l after 8 weeks of dietary intervention. The patients were treated with either fluvastatin 40 mg daily or with the combination of fluvastatin (20 mg daily) and micronized fenofibrate 200 mg daily. Mean values of total and LDL cholesterol decreased by 19.3% and 29.7% respectively after fluvastatin treatment and by 21.5% and 29.1% respectively after the combination of fluvastatin and fenofibrate treatment. The differences between the treated groups were not significant. Mean values of HDL-cholesterol increased significantly more after the combination of fluvastatin and fenofibrate than after fluvastatin monotherapy (26% vs. 9.9%). The mean values of triglycerides decreased significantly more after the combination treatment than after fluvastatin monotherapy (-40.2% vs. 19.7%). The treatment in both groups was well tolerated and no signs of myopathy were observed in any patient. The study was discontinued in 1 patient due to the increase of liver enzymes. The most frequently observed side effects were minor gastrointestinal symptoms, which were more frequent in patients treated by the combination of fluvastatin and fenofibrate. Thus our results demonstrate that the combination of fluvastatin and fenofibrate is an effective and safe treatment option for patients with coronary heart disease and mild to moderate combined hyperlipidemia if a more radical lowering of triglycerides and increase of HDL cholesterol is desired. PMID- 11045183 TI - [Pregnancy in patients with hereditary spherocytosis]. AB - The influence of pregnancy on the course of hereditary spherocytosis was investigated in 21 women during their 44 pregnancies. Fourteen pregnancies were followed up directly, 30 were evaluated from anamnestic data. In the majority of investigated women with hereditary spherocytosis pregnancy caused no problems. When complications developed, they were not serious as a rule. Only about one third of pregnancies in non-splenectomized women developed anaemia or anaemia deteriorated. In the latter enhanced haemolysis participated. In splenectomized patients the incidence of complaints was minimal. PMID- 11045182 TI - [Serum levels of zinc, copper and selenium in patients with Wilson's disease treated with zinc]. AB - Zinc administered on a long-term basis in excess to patients with Wilson a disease blocks in a significant way copper absorption from the gut, prevents its accumulation and toxic action in the organism. The authors investigated the effect of its long-term administration on the plasma concentration of copper, zinc, and selenium, on the superoxide dismutase activity in red blood cells and glutathione peroxidase activity in whole blood. In seven patients with Wilson a disease treated with zinc sulphate, 136 mg of elemental zinc for 1.5 years (18 months), the authors assessed the plasma concentration of zinc, copper, selenium and ceruloplasmin, the activity of superoxide dismutase in red blood cells, the activity of glutathione peroxidase in whole blood and the urinary excretion of zinc and copper in 24 hours. Envisaged findings with regard to the diagnosis of the investigated patients and their treatment: elevated plasma zinc concentration and increased urinary excretion, reduced copper and ceruloplasmin plasma concentration and increased urinary copper excretion. The authors recorded also a significantly elevated selenium plasma concentration and a significantly higher concentration of superoxide dismutase in red blood cells (p < 0.05). The increase of the glutathione peroxidase activity in whole blood in the investigated patients was not significant (p < 0.05). Changes in the values of the investigated parameters in patients with Wilson s disease treated on a long-term basis with zinc indicate the possible mutual interaction of zinc with other trace elements with an impact on the activity of the corresponding metalloenzymes, i.e. in the sphere in antioxidant systems. PMID- 11045184 TI - [Pregnancy after kidney transplantation]. AB - In the period 1966-1997 renal allografting was performed in 1746 recipients, 244 of whom were women in fertile age. In 32 of them 45 pregnancies were registered. 29 of them (64%) resulted in abortion, which was spontaneous in 4 and medically advised in 25. There were 16 labours, 4 of them free of any complications; of the latter, hypertension was the most frequent one (8). Of the former, 13 were solved by caesarean section and 3 were vaginal deliveries. Of 15 live babies 7 were immature (one died 1.5 year later of renal failure due to microcystic kidneys). No unfavourable effect of pregnancy on prognosis and on long-term graft function was found. PMID- 11045185 TI - [Conn's syndrome and severe arrhythmias]. AB - In the submitted case-history the authors describe Conn's syndrome. A solitary cortical adenoma was involved manifested at first by so-called "lone" atrial fibrillation (idiopathic without organic causes). The trigger factor of the life threatening ventricular tachycardia were variable values of serum potassium and treatment of atrial fibrillation by anti-arrhythmic drugs. PMID- 11045186 TI - [Endothelial dysfunction and arterial hypertension]. AB - The endothelium lines all blood vessels in the human body, it is the basic structure which ensures the action of substances circulating in the blood stream on the vascular wall. It is an organ the sound state of which is essential for the physiological function of the vascular system. Its impaired function is a basic factor in the genesis and development of vascular disease. Under physiological conditions the endothelium has antiadhesive and antithrombotic properties, it produces vasoactive substances, prevents the penetration of circulating substances and formed elements across the vascular wall, and via adhesion molecules it participates in the interaction with cells in the circulation. Risk factors of cardiovascular diseases such as hypertension, hyperlipidaemia, hyperglycaemia, smoking damage the function of endothelial cells and cause the development of endothelial dysfunction. In patients with arterial hypertension endothelial dysfunction is characterized by an impaired endothelium dependent relaxation, increased adhesion and permeability of endothelial cells, structural changes of the vascular wall. When the endothelium is damaged by released cytokines an increased expression of adhesion molecules occurs, adhesion and migration of inflammatory cells across the vascular wall. Cytoadhesion molecules are released from the surface of the endothelium into the circulation where the rise of their plasma levels can serve as a marker of endothelial damage. Endothelial dysfunction in hypertonic subjects contributes in a significant way to the development and progression of chronic vascular disease- atherosclerosis. Improvement of the damaged endothelial function is therefore at present a desirable therapeutic objective in the treatment of hypertension. PMID- 11045187 TI - [New cytokines and their role in supportive care]. AB - Several cytokines stimulating hematopoiesis, mainly lineage restricted, are already widely used in supportive care to correct myelosuppression or anaemia (GM CSF, G-CSF, EPO). The new growth factor are tested in preclinical or clinical studies to abrogate other anti-cancer therapy side-effects (thrombocytopenia, mucositis etc.). IL-3 has been shown to have only limited effect on neutrophils and platelets production respectively. IL-6 and IL-11 have been tested to improve thrombocytopenia and mucositis (IL-11). Thrombopoetin (TPO, c-mpl) is tested in clinical trials and shows very strong effect on platelet counts. Stem cell factor (SCF) has shown to improve progenitor cell mobilisation, particularly in combination with other cytokines. The new promising factor, FLT-3 ligand, combines effect on hematopoiesis with effect on dendritic cells generation. The new group of synthetic cytokines (daniplestim, myelopoetin, promegapoetin and progenipoetin) is now tested in preclinical and clinical studies. Mucositis could be influenced by new keratinocyte growth factor (KGF), which is now in phase I trials. PMID- 11045188 TI - [Drug therapy of esophageal varices hemorrhage in portal hypertension]. AB - The author discusses the etiopathogenesis of portal hypertension and possibilities how to influence it during treatment of acute haemorrhage from varicosities and how to implement primary and secondary prevention. In treatment of acute haemorrhage the author recommends terlipresin, 1 mg every 4 hours. In primary and in particular in secondary prevention he emphasizes the necessity of early administration of beta-blockers. PMID- 11045189 TI - [Renaissance of fluorides and calcium in the treatment of osteoporosis]. AB - After the initial enthusiasm for fluorides in the treatment of osteoporosis there was a regression after some time and a "campaign" against fluorides. Recently in particular after the introduction of monofluorophosphate instead of NaF there is a revival of the use of fluorides in the treatment of osteoporosis, along with Ca, vitamin D and exercise as the classical combination of four. This can be supplemented by HRT in women, thyrocalcitonin, alendronate and other drugs with individual consideration of the optimal combination of these drugs. Also in future fluorides remain the foremost representative of the group of agents stimulating new formation of bone. PMID- 11045190 TI - [The concept of internal medicine and its subspecialties at the end of the 2nd millennium. An open letter to members of the Czech Internal Medicine Society]. PMID- 11045191 TI - The making of an internist: an European view. PMID- 11045192 TI - [What will internal medicine look like in developed countries in the 21st century?]. PMID- 11045193 TI - The role of surgery in asymptomatic lumbosacral spinal lipomas. PMID- 11045194 TI - An introduction to the changes in gene expression that occur after cerebral ischaemia. AB - Ischaemia is the final common pathway of brain cell death following a variety of insults. We consider the effect of cerebral ischaemia on gene expression, concentrating on immediate early genes, those encoding heat shock proteins, growth factors, cytokines, adhesion molecules, nitric oxide synthase and proteins involved in programmed cell death. We conclude that the changes in gene expression resulting from ischaemia are important determinants of neuronal and glial survival. In the future it may become possible to manipulate gene expression to limit the extent of damage arising from cerebral ischaemia. PMID- 11045195 TI - Primary cerebral non-Hodgkin's lymphoma: problems with diagnosis and development of a protocol for management. AB - The prognosis for primary cerebral non-Hodgkin's lymphoma has improved markedly in the under-70 age group with the introduction of combined chemotherapy and radiotherapy. A review of our patients has revealed difficulty in obtaining definitive histology initially in 21% (9/43). We investigated the role of corticosteroids in these patients and found an idiosyncratic response in that there was no significance either of the cumulative dose (p > 0.424) or the length of treatment (p > 0.453) with the observed regression of lymphoma with corticosteroids. In order to avoid indiscriminate corticosteroid administration we have formulated a management protocol the entry point for which is a patient with enhanced computered tomography appearances of primary cerebral lymphoma. PMID- 11045196 TI - Effect of photodynamic therapy on recurrent pituitary adenomas: clinical phase I/II trial--an early report. AB - Pituitary adenomas, although histologically benign, are not always curable by surgery alone, principally because of dural infiltration, as well as their peculiar anatomical location. Radiotherapy has been employed as an adjuvant therapy to address residual disease with favourable results. This approach is, however, not without side effects, and it cannot be repeated. We are therefore investigating the effectiveness of photodynamic therapy (PDT) on recurrent pituitary adenomas in humans. This study details the protocol applied to 12 patients with recurrent pituitary adenomas, which involved systemic administration of photosensitizer (Photofrin) followed, after a period of 24-48 h, by intraoperative illumination of the tumour bed using 630 nm laser light. The primary end points were visual, endocrine and radiological improvement. The incidence of side effects was also monitored. The longest follow-up is 2 years. Most patients suffering from visual acuity or field defects have shown improvement when followed for 12 months or more. Three patients showed complete recovery of their visual fields. All those who presented with functional adenomas have shown reduction in their hormone levels. Tumour volume, relative to the preoperative size, was 122, 87, 66, 60 and 46% at 4 days, and 3, 6, 18 and 24 months, respectively. One patient developed severe skin photosensitization due to early exposure to direct sunlight and three others displayed minor skin reactions. There was no treatment-related mortality or morbidity. One patient (operated transcranially) developed hemiparesis postoperatively, which recovered completely. We think this is unrelated to the treatment. This prospective study demonstrates that PDT may be safely applied to the pituitary fossa by the trans sphenoidal route and indicates the need for a randomized, controlled trial in order to establish its therapeutic potential. PMID- 11045197 TI - Traumatic cranial empyemas: a review of 55 patients. AB - A 15-year (1983-1997) review of our unit's computed tomographic experience with traumatic cranial empyema (TCE) is reported. Fifty-five patients with documented history and clinical evidence of neurotrauma with secondary cranial empyema at surgery were identified. The clinical records and CT scans were analysed. TCE [four extradural and 51 subdural collections (SDE)] accounted for 7.86% of the total cranial empyemas seen during the study period. Most of the patients were young males (44 patients) and neurological deficits on admission were found only in the SDE group. Forty-one of 53 patients presented with septic compound skull fractures. Fifty-four patients had urgent surgical drainage. Eighty per cent of patients experienced a good outcome (GOS 4 or 5). A morbidity of 16.4% (including postoperative seizures) was noted and eight patients died (mortality rate 14.5%). Urgent surgical drainage, removal of osteitic bone, wound debridement and high dose intravenous antibiotic therapy form the mainstay of treatment. PMID- 11045198 TI - Spinal intramedullary ependymomas: surgical results and immunohistochemical analysis of tumour proliferation activity. AB - This study analysed the outcome of 35 consecutive patients with intramedullary ependymoma who underwent a radical surgical resection between 1983 and 1996. The median age of patients was 47 years. Tumour location was cervical in 22 patients, cerviothoracic in eight and thoraco lumber in five. The proliferative activity of 24 spinal and 14 intracranial ependymomas were determined by MIB-1 immunolabelling. Total removal of tumour was achieved in 26 patients and subtotal removal was performed in nine patients of whom six received postsurgical radiation therapy. Twenty-nine patients out of 35 could be followed over 3 years (follow-up periods: 168-36 months: mean 70). Moreover many patients could be followed over 4 years. In 27 of those 29 patients, the neurological symptom was stabilized or improved. The proliferation indices of spinal ependymomas were significantly lower than those of intracranial ependymomas. Tumour regrowth occurred only in a young patient 29 months after a subtotal removal of the tumour which was not treated with irradiation and showed a high proliferation index on the second operation. PMID- 11045199 TI - Dysphagia secondary to iatrogenic hyoid bone fracture. AB - Dysphagia as a result of an iatrogenic fracture of the hyoid bone is presented. Review of the world literature reveals this phenomena to be previously unreported following surgery. PMID- 11045200 TI - Minimalist placement of external ventricular drains. A quick, safe, cheap and cosmetic procedure. AB - We report the use of a twist drill method for placement of external ventricular drains and ventriculoperitoneal shunts. We have found this method to be efficient and safe, and provides an excellent cosmetic effect. PMID- 11045201 TI - Neurosurgery without shaving: indications and results. AB - The objective of the investigation was to prevent the stigmatizing effect of a totally or partially shaved head with openly visible signs of a head operation, easing the reintegration of the patient into his daily life. After extended surface cleaning with a colourless, regular skin disinfection liquid (undyed isopropanol/dibrom/propylenglycol solution) the hair was combed apart from the incision line before draping. Wound closure was performed as usual, taking care to remove meticulously hair from the wound. To aid closure we made use of a 20% chlorhexidine jelly that holds the hair away from the incision. A neomycin ointment served for sealing the wound surface, no further dressing being used. After 215 cranial neurosurgical operations, among them 63 for tumours, 33 stereotactic procedures 18 shunt, 55 for torticollis and 46 other operations performed without shaving the hair, we saw one wound infection (0.5%). This percentage corresponds very well to our general infection rate of 0.6%. All patients very much appreciated the offered opportunity and the result. If the objective is to give patients a psychological advantage by preventing a partially bald head we think one can safely refrain from the shaving procedure without risking a higher infection rate. PMID- 11045202 TI - Cervical intramedullary schwannoma: complete excision using the KTP laser. AB - Intramedullary cervical spinal cord schwannomas are rare tumours and complete excision is often an elusive goal. The use of the KTP laser to accomplish complete excision has not been reported previously. Postoperatively, our patient had no additional deficit and after 1 year has made a good recovery. This case further highlights the difficulty in interpretation of intraoperative biopsy specimens. PMID- 11045203 TI - Self-inflicted head trauma using a captive bolt pistol: report of three cases. AB - Three cases of self-inflicted head trauma using a captive-bolt pistol are described. This is a rarely reported phenomenon and presents with an unique pattern of brain injury. PMID- 11045204 TI - Thoracic radiculopathy caused by a myodil cyst. AB - We report the case of a myodil cyst causing a thoracic radiculopathy in a patient who had undergone a myelogram 30 years previously. Although myodil is no longer used, sequelae can continue to be seen for many years. PMID- 11045206 TI - Brain stem tuberculosis in children. AB - Tubercular meningitis and intracranial tuberculomas are the two frequent manifestations of neurotuberculosis with their variable incidence in different countries. Brain stem tuberculomas are even more unusual, accounting for 2.5-8% of all intracranial tuberculomas. We present here six paediatric cases of brain stem tuberculosis, where well-formed tuberculomas were demonstrated in five symptomatic cases; however, only a hypodense ill-defined lesion was found in the brain stem of one child who presented with features of tubercular meningitis. Three of these children had other associated tuberculomas in their brain parenchyma as well. All the six cases, however, showed the involvement of 3rd, 6th & 7th cranial nerves, in various combinations, at the time of initial presentation. Only two children developed hydrocephalus and required CSF diversion. All responded well to anti-tubercular chemotherapy, though two children developed toxicity to the therapy, which recovered with drug manipulation for about 2 months. Recovery was full without residual deficit in all the cases. Surgical excision of brain stem granulomas was not indicated in any case. PMID- 11045205 TI - Spontaneous regression of a solitary cerebral metastases in renal carcinoma followed by meningioma development under medroxyprogesterone acetate therapy. AB - A case of regression of a probable cerebral metastasis of a hypernephroma after nephrectomy and hormone therapy is presented. The patient subsequently developed a meningioma after therapy with medroxyprogesterone acetate. A relationship between meningioma growth and sex hormones has been documented, but little is known about the effect of hormone therapies on tumour growth. PMID- 11045207 TI - Craniovertebral junction malformation and rotational occlusion of the vertebral artery. AB - An unusual case of rotational occlusion of a non-dominant vertebral artery (VA) is presented. This clinical syndrome is associated for the first time with a complex craniovertebral junction malformation. Rotational occlusion of the VA is usually asymptomatic, vertebro-basilar insufficiency has been rarely observed in cases of involvement of a dominant VA, and concomitant occlusion or hypoplasia of the contralateral artery. Diagnosis of rotational VA occlusion is based on haemodynamic demonstration of the effects of head rotation. The importance of the awareness of this syndrome is stressed. PMID- 11045208 TI - Cerebellar astrocytoma in an adult with extensive leptomeningeal spread. AB - A rare case of a benign cerebellar astrocytoma in an adult is reported. This tumour had an extensive leptomeningeal spread in the cerebellar convexity, basal cisterns and spinal cord. The most unusual and radiologically demonstrated extension was also into the sella and Meckel's cave. The case is discussed and the literature is briefly reviewed. PMID- 11045209 TI - Congenital spinal astrocytoma: how favourable is the long-term outcome? AB - An infant born with a low grade cervical astrocytoma and the adverse neuro developmental outcome despite successful resection of the tumour are described. This case adds to the literature on this rare condition and the need for improvement in the management of these infants. PMID- 11045210 TI - Tentorium cerebelli subdural haematoma complicating subarachnoid haemorrhage. AB - Acute subdural haematoma (SDH) as a result of aneurysmal rupture is a rare event. We report a tentorium cerebelli SDH, secondary to a ruptured aneurysm which we believe is unique in the literature. This report also gives further support to the theory that a sentinel bleed causes inflammation and adhesion between the aneurysm and arachnoid as the underlying pathology in these unusual subdural haematomas. Since the first report of acute SDH from a ruptured aneurysm by Hasse in 1855, only about 150 further cases have been reported in the literature. We describe a further case, but at an as yet unreported site with radiological evidence of a sentinel bleed. PMID- 11045211 TI - Spontaneous resetting of a Codman-Medos programmable shunt. PMID- 11045212 TI - Ruptured transverse ligament: an injury that is often forgotten. PMID- 11045213 TI - Dysembryoplastic neuroepithelial tumour. PMID- 11045214 TI - A rare Behr. PMID- 11045215 TI - Bone window CT evaluation on nasal cavity for transsphenoidal reoperations: a sequel to a previous publication. PMID- 11045216 TI - Spinal angiolipoma during pregnancy: review of the literature. PMID- 11045217 TI - Psychiatric treatments: an exciting new century. PMID- 11045218 TI - The use of sildenafil in heart disease. PMID- 11045219 TI - Familial ovarian malignancy. AB - Familial ovarian cancer is a subset of ovarian cancer where there appears to be a definable genetic link in families. It accounts for less than 5% of cases of ovarian cancer seen at present. The main genes responsible, BRCA1 and BRCA2, are discussed. Screening and the prophylactic measures for women identified having a genetic trait are discussed. PMID- 11045220 TI - Amniotic fluid embolism: the UK register. AB - Amniotic fluid embolism is rare but is a significant cause of maternal death. No clear risk factors seem to be identifiable from previous cases. A register has been established in the UK to look at possible therapies. PMID- 11045221 TI - Vault haematoma and febrile morbidity after vaginal hysterectomy. AB - Vaginal hysterectomy is associated with significant risk of vaginal vault haematoma (40%) and febrile morbidity (30%). Numerous interventions have been used to try and avoid these complications but very few have been shown to be effective. PMID- 11045222 TI - The modern geriatric day hospital. AB - Geriatric day hospitals have played a major role in the rehabilitation of older people, although the evidence base has proved thin. As the provision of geriatric medicine changes, they need to develop new roles such as responding to subacute crises, providing specialist services and ensuring comprehensive geriatric assessment before long-term care. PMID- 11045223 TI - A simple guide to nystagmus. AB - Nystagmus is a potentially mind-boggling subject, as over forty different types are recognized. However, by classifying the various types into a small number of categories, the clinician can make sense of them, and so make use of a very important clinical sign. PMID- 11045224 TI - Subfertility following treatment for childhood cancer. AB - The majority of children with cancer can now expect to be cured. Recent advances in assisted reproduction have focused attention on the long-term fertility outcome for these survivors. This article summarizes the current state of our knowledge about who is at risk of subfertility after childhood cancer and discusses possibilities of fertility preservation that are currently being researched. PMID- 11045225 TI - Refining the use of inhaled steroids in asthma. AB - Inhaled corticosteroids remain the most significant aspect of asthma therapy. An increasing number of drugs, devices and formulations are available to health-care professionals. This article aims to address the goals of inhaled steroid therapy and asks how well the available options meet these goals. PMID- 11045226 TI - Acute respiratory distress syndrome in adults. AB - The acute respiratory distress syndrome (ARDS) can arise from a range of predisposing insults. Mortality rates are high for patients with ARDS and survivors require extended and expensive intensive care treatment. This article presents evidence that implicates the production of toxic and damaging reactive oxygen species in the pathogenesis of ARDS. PMID- 11045227 TI - PRHOs in primary care: what is happening out there? AB - Preregistration house officer (PRHO) placements in general practice were introduced throughout Britain in August 1998. This paper describes an evaluation of PRHOs in primary care rotations in South Thames during 1998-99. There are important messages for both educational supervisors and undergraduates considering a PRHO in primary care rotation. PMID- 11045228 TI - A surgeon in New York: a view of the American internship. AB - A young surgeon describes life as an intern in New York. Well-supervised hands-on operative experience and an assured place on a training programme for most residents compensate for low pay and gruelling hours of work. PMID- 11045229 TI - The madness of Nietzsche: a misdiagnosis of the millennium? AB - This article represents a personal discussion about Nietzsche's mental illness, which formed part of a larger paper 'The masks of Nietzsche and eternal return of the repressed'. This was presented at the 6th Annual Conference of The Friedrich Nietzsche Society, September 1996, Manchester UK, as reported by Nussbaumer-Benz (1998). PMID- 11045230 TI - False diagnosis of a catecholamine-secreting paraganglioma in a patient with hypertension and depression. PMID- 11045231 TI - Delayed diagnosis of HIV in patients with reactive lymphadenopathy. PMID- 11045232 TI - Penile cellulitis. PMID- 11045233 TI - Giant urethral calculi. PMID- 11045234 TI - Use of Viagra by patients with heart disease. PMID- 11045235 TI - Does smoking help spread MRSA? PMID- 11045236 TI - Confidence--accuracy relations for real and suggested events. AB - Participants completed a multiple item, multiple event version of a traditional misinformation procedure, as well as a battery of individual difference measures. The relation between memory accuracy and self-reported confidence was assessed through the comparison of items involving misinformation and items not involving misinformation. Selected individual differences in the confidence-accuracy relation were also examined for items that did and did not involve false post event information. Results indicated significant differences between the measure of the confidence-accuracy relationship for misinformation and non-misinformation items. Several significant, although weak, individual difference correlates of the confidence-accuracy relations were also found. PMID- 11045237 TI - When the smile is a cue to familiarity. AB - The question discussed in the two following experiments concerns the effect of facial expressions on face recognition. Famous and unknown faces with neutral or smiling expression were presented for different inspection durations (15 ms vs 1000 ms). Subjects had to categorize these faces as famous or unknown (Experiment 1), or estimate their degree of familiarity on a rating scale (Experiment 2). Results showed that the smile increased ratings of familiarity for unfamiliar faces (Experiments 1 and 2) and for famous faces (Experiment 2). These data are discussed in the framework of current face-recognition models and are interpreted in terms of social value of the smile. It is proposed that the smiling bias found here acts at the level of the decision process. PMID- 11045238 TI - Deaf and hearing adults' recollections of childhood and beyond. AB - Whereas the average age of earliest reportable personal memory among adults is 3 to 3 1/2, there is considerable individual and group variability in the age of earliest autobiographical memory. Some of the variability is thought to be attributable to differential narrative socialisation. In the present research we tested the hypothesis that by virtue of later exposure to language, individuals born deaf to hearing parents will have earliest memories from later in life, relative to hearing individuals. The average age of single earliest identifiable memory for adults who are deaf and adults who are hearing did not differ. Nevertheless, adults who are deaf were found to have less dense representations of early autobiographical memories and to include in their narrative reports fewer categories of information, including visual-spatial information, relative to hearing adults. Participants' ratings of their memories on a number of dimensions were found to have low utility in predicting the content of autobiographical reports from both early and later in life (i.e., after age 10 years). PMID- 11045240 TI - Ordering autobiographical experiences. AB - This study examined individuals' memory for the temporal order of autobiographical events and for the components that constitute autobiographical events. Study 1 measured performance on an across-event ordering task that involved the chronological arrangement of cards that displayed event labels. Results indicated poor ordering ability across events, but a reasonable ability to order clusters of events. Study 2 compared within-event and across-event ordering using computer-presented digital photographs. Participants were better at ordering the photographs in their own across-event trials than in their within event trials. The results are discussed in terms of the retrieval of temporal information under within- and across-event conditions. PMID- 11045239 TI - Prospective and retrospective memory in normal ageing and dementia: a questionnaire study. AB - Frequency of prospective memory and retrospective memory failures was rated on a 16-item questionnaire by 862 volunteers, from five groups: patients with Alzheimer Disease (rated by carers), carers of Alzheimer Disease patients, elderly, young, and a group of married couples. Reported memory failures were highest for Alzheimer Disease patients, and lowest for carers, with elderly and young controls in between. More prospective memory than retrospective memory failures were reported in all groups, although the difference was small for Alzheimer Disease patients who were rated near ceiling for both. Prospective memory failures of Alzheimer Disease patients were reported as more frustrating for carers than retrospective memory failures; prospective memory and retrospective memory failures frustrated Alzheimer Disease patients equally. Data from the couples indicated that there were no biases resulting from rating on behalf of someone else. These results suggest that: (1) normal ageing has no greater effect on self-reported retrospective memory than prospective memory failures, (2) the relatively small number of memory failures reported by carers may result from comparing themselves with the Alzheimer Disease patients in their care, and (3) prospective memory failures have a greater impact on the lives of the carers and are therefore more likely to be reported as early indicants of the disease. PMID- 11045241 TI - Conscious and unconscious influences of memory: temporal dynamics. AB - Changes in the conscious and unconscious influences of memory over time were assessed in two experiments by using a variant of the process-dissociation procedure. In both experiments, performance on a stem-completion task was measured under both inclusion and exclusion instructions. Across the two experiments, there were four different retention intervals: 2 minutes, 2 days, 2 weeks, and 2 months. The results indicated that conscious influences decreased systematically across retention interval. In contrast, unconscious influences of memory in the absence of conscious influences increased between 2 minutes and 2 days, and then remained relatively stable from 2 days to 2 weeks to 2 months. These results stand in apparent contrast to those of McBride and Dosher (1999), which showed equal rates of forgetting for conscious and unconscious influences of memory on performance. The implications for models of the relation between conscious and unconscious influences of memory on performance are discussed. PMID- 11045242 TI - Disruption of comprehension by the meaning of irrelevant sound. AB - This study investigates the claim that the disruption of comprehension by irrelevant sound is qualitatively different from that of short-term memory for order. Both meaningful and meaningless speech disrupted the comprehensive aspect of the task, but the effect of meaningful speech was significantly greater. Both rehearsal and semantic processing, which are involved in reading comprehension, seem to be susceptible to disruption by irrelevant meaningful speech. The study provides some evidence to suggest that the presence of meaning in the irrelevant sound in creases disruption of performance in cognitive tasks that also call upon processing of meaning. PMID- 11045243 TI - An optometrist's role of co-management in a hospital glaucoma clinic. AB - In recent years, optometrists have started to work in some clinical roles in hospital medical clinics. This paper outlines the current working practice in a typical glaucoma clinic at Moorfields Eye Hospital NHS Trust and goes on to establish the ability of an optometrist working with a team of ophthalmologists. In this study, 54 patients (n = 108 eyes) were recruited and clinically assessed by an optometrist. Subsequently, a research fellow (i.e. ophthalmologist) assessed the same patients independently. The results were compared in order to determine the accuracy of the optometrist's evaluation and proposed management. This paper considers the costs of employing personnel by reviewing the salaries of optometrists and medical assistants. Although nurses have started to work alongside ophthalmologists, and orthoptists may be considering such work, it is not within the scope of this paper to consider either of these groups. If optometrists are to become an accepted part of a glaucoma clinical team, consideration needs to be given to the training and experience they receive. PMID- 11045244 TI - Oral and silent reading performance with macular degeneration. AB - Previous studies have shown that reading rate for very large print (6 degrees, 1.86 logMAR character size) is a strong predictor of oral reading rate with low vision devices (LVDs). We investigated whether this would apply using large print sizes more readily available in clinical situations (e.g. 2 degrees, 1.4 logMAR), for subjects with macular degeneration. We assessed rauding rates--reading for understanding. A combination of near word visual acuity and large print reading rate (without LVDs) provided the best prediction of oral rauding rates (with LVDs). However, near word visual acuity alone was almost as good. Similarly, silent rauding rate was predicted best by near word visual acuity alone. We give near visual acuity limits as a clinical guide to expected oral and silent reading performance with LVDs for patients with macular degeneration. PMID- 11045245 TI - Detection, aetiology and management of conjunctival intraepithelial neoplasia. AB - Conjunctival intraepithelial neoplasia (CIN) are considered to be the third most common ocular tumour and the most common tumour of the ocular surface. Due to their malignant potential, they must be carefully differentially and promptly treated. A recurrence rate of approximately 30% leads to the need for monitoring of patients even after successful treatment. This article presents several cases of CIN and reviews their histopathology, aetiology, appearance, differential diagnosis and management. Typical patient presentations and prognoses are also discussed. PMID- 11045246 TI - Iris color and age-related changes in lens optical density. AB - PURPOSE: Epidemiologic evidence indicates that dark iris color increases risk of age-related cataract. No information is currently available, however, on the effects of iris color on the lens prior to cataract development. In this study, we relate iris color to lens optical density (OD) in individuals without frank cataract. METHODS: 90 subjects with blue or green irises (light color) were compared with 87 subjects having hazel, brown, or black irises (dark color). Lens OD was measured psychophysically by comparing scotopic thresholds obtained at 410 (measuring) and 550 nm (reference). Stimuli were presented in Maxwellian view. RESULTS: The groups with light and with dark iris color did not differ significantly in smoking habits, dietary patterns, or age. Despite other similarities between the groups, lens OD was significantly (p < 0.024) higher in the group with dark irises. The higher OD of the dark iris group was due to differences in the older subjects (> 45 years, p < 0.005), rather than the younger subjects (20-45 years) who showed no differences in lens OD. CONCLUSION: Our data indicate that iris pigmentation may be directly related to age associated increases in lens OD. PMID- 11045247 TI - Pulse-duration threshold functions obtained at the MWS/LWS cone antagonistic locus. AB - Pulse-duration threshold functions were determined for spectral increments in the region of the Sloan notch (580 nm) as well as for 440 and 640 nm and white increments flashed on a spatially coincident white background. While increments of 440 and 640 nm elicited traditional pulse-duration functions with decreasing thresholds as the duration increased, functions obtained with 580 nm and white increments manifested a minima at about 50 ms. These data support the notion that stimuli in the region of Sloan's notch can be detected by an achromatic mechanism that may integrate sub-threshold ON and OFF responses, thereby maximizing detection sensitivity. PMID- 11045248 TI - Further analysis of the size and shape of cells obtained by impression cytology from the exposed portion of the human bulbar conjunctiva. AB - PURPOSE: To assess whether the size of the cells obtained by conjunctival impression cytology can be quantitatively assessed by measurement of the longest dimension of the cells. METHODS: Under topical benoxinate anaesthesia, cells were removed from the normally exposed nasal bulbar conjunctival surface using a 0.4 micron pore diameter filter (Biopore filter; type Millcell-CM). The filters were stained with haematoxylin after ethanol denaturation, photographed at 40 x magnification, and 35 mm slides prepared. An optical overlay method was used to outline the borders of sets of 30-35 contiguous cells on each image. The cell area, longest and shortest dimensions were measured by planimetry to an accuracy of +/- 3%. RESULTS: Analyses of 20 sets of samples, from individuals aged 21 to 48 years and without clinically significant ocular surface disease, revealed a median cell area of 133 micron 2 (n = 621, range 46-1602 micron 2; average 212 micron 2), a median longest dimension of 13.9 microns (range 6.6-68.8 microns; average 16.9 microns) and a shorter dimension of 10.0 microns (range 4.7-43.0 microns; average 11.9 microns); the distributions of values indicated bimodality. Most cells had a long:short ratio (L:S ratio) value between 1.00 and 1.80, but 11.9 +/- 6.1% of the cells had L:S ratios between 1.80 and 4.60. The overall relationship between the longest dimension and the area of the cells was nonlinear, with cells having larger L:S ratios having disproportionately smaller areas. CONCLUSIONS: The superficial conjunctival cells are small, and their longest dimensions are systematically related to area. Analyses of cell shape indicate further possible ways of identifying different cells on the bulbar conjunctiva. Compared to literature values, there is a substantial overlap in longest dimensions of the conjunctival cells with those of cells that can be collected off the corneal surface. This means that superficial conjunctival and corneal cells cannot be distinguished simply on the basis of measurements of the long dimension. PMID- 11045249 TI - Intraocular pressure and pulse rate measurements by the OBF tonograph--comparison to reference instruments. AB - The purpose of this study was to evaluate the accuracy of intraocular pressure (IOP) and ocular pulse rate (OPR) measurements obtained by the Ocular Blood Flow (OBF) tonograph (OBF Labs, Wiltshire, UK). Measurements of IOP and OPR by the OBF tonograph were compared to those of reference instruments. For IOP evaluation, measurements were obtained on patients with normal and abnormal pressures using the OBF tonograph and the Goldmann applannation tonometer in random alternate order. For the OPR evaluation, measurements were obtained using the OBF tonograph with simultaneous heart rate monitoring by ECG on patients with normal IOP. The validity of the OBF tonograph measurements was quantified in terms of 95% limits of agreement and their relationships to measurements by reference instruments was determined by linear regression analyses. 102 patients were recruited for IOP measurements. Mean IOP obtained by the Goldmann tonometer was 20.7 mmHg (7-42 mmHg, SD 6.98) whilst mean IOP obtained by the OBF tonograph was 20.1 mmHg (8.1 40.2, SD 6.1). Goldmann IOP and OBF tonograph IOP readings were well correlated (r = 0.945). Analysis of the difference in IOP measurements between two instruments (tonograph minus Goldmann tonometer) showed the mean bias to be 0.26 mmHg (-7.8 to +6.1 mmHg) and the 95% limits of agreement to be -4.35 to +4.87 mmHg. Agreement between two instruments appeared to be dependent on the IOP; at IOP lower than 20.6 mmHg there was an overall tendency for the tonograph IOP to be higher than Goldmann IOP and vice versa when IOP was above 20.6 mmHg. 47 patients were recruited for OPR and ECG measurements. Mean pulse rates were 74.8 beats per min (mean 43-110) by ECG and 73.9 beats per min (43-110) by tonography. Analysis of the difference in pulse rate between instruments (tonograph OPR minus ECG pulse rate) against the average pulse rate showed the mean bias to be -0.8 beats and the 95% limits of agreement to be between -7 to +5 beats. Ocular pulse rate values obtained by the OBF tonograph were very accurate when compared to ECG pulse rate. This indicates that there is unlikely to be a systematic lag in continuous ocular pulse waveform recording. Intraocular pressure measurements by the OBF tonograph correlated very well with Goldmann readings over a wide range of pressures and should be valid in the clinical setting. PMID- 11045250 TI - Does ocular refraction change with ambient temperature? PMID- 11045251 TI - Outcome for continuous ambulatory peritoneal dialysis patients is not predicted by peritoneal permeability characteristics. AB - For the present study, we investigated the peritoneal transport of fluid and solutes and the clinical outcomes of 44 continuous ambulatory peritoneal dialysis (CAPD) patients with various peritoneal transport characteristics. Based on 24 hour urine and dialysate collections and 4-hour dwell studies [peritoneal equilibration test (PET)], the patients were divided into two transport groups by dialysate-to-plasma ratio of creatinine at 240 minutes (D/PCr240). The groups consisted of 21 high transporters (D/P = 0.81; mean age: 63.9 +/- 8.2 years) and 23 patients of other transport types (D/P < 0.81; mean age: 67.1 +/- 7.3). Mean CAPD duration was 57.14 +/- 30.4 months and 39.14 +/- 30.4 months respectively (p = 0.07). Estimations were made of weight, body surface area (BSA), percent body water, plasma albumin (PA), Kt/Vurea, weekly creatinine clearance (TCCr), fluid removal, residual renal function, and normalized protein catabolic rate (nPCR). The results showed that high transporters had statistically significant, lower values for: (1) peritoneal fluid (p = 0.02); (2) estimated glomerular filtration rate (GFR: 0.5 +/- 0.77 mL/min vs 2.15 +/- 2.2 mL/min, p = 0.002); and (3) nPCR (0.66 +/- 0.16 g/kg/day vs 0.84 +/- 0.23 g/kg/day, p = 0.003). No statistically significant differences were observed with regard to the other parameters (BSA, PA, Kt/Vurea, TCCr). Cumulative survival rates at two and five years were 90% and 70% for all patients. No statistically significant difference was seen when comparing the survival curves of high transporters and patients of other transport types (p = 0.33, Cox's F-test). In conclusion, we saw no clear evidence that higher peritoneal permeability negatively influences clinical outcome. Other comorbid factors may be related in a more important way to the survival rate for CAPD patients. PMID- 11045252 TI - Amino-acid-based dialysis solution changes leptinemia and leptin peritoneal clearance. AB - In continuous ambulatory peritoneal dialysis (CAPD) patients, nutritional parameters, appetite, and transperitoneal solute movement can be modified by treatment with amino-acid-based dialysis solution (AADS). Because leptin is involved in energy expenditure and appetite regulation, we decided to examine the influence of AADS on serum and dialysate leptin concentrations. We prospectively evaluated AADS influence on leptinemia and peritoneal transport indices in CAPD patients. Nine clinically stable patients (7 males, 2 females), mean age 55.4 +/- 10.5 years, who had been treated with CAPD for 6.1 +/- 5.8 months, were studied. Examinations were conducted before treatment with 1.1% AADS (period I), after 3 months of AADS administration (period II), after 6 months of AADS administration (period III), and at 3 months after AADS discontinuation (period IV). The primary outcome measure was concentration of leptin in serum and dialysate. Secondary measures included anorexia incidence, nutrient intake, and nutritional parameters. Dialysate-to-plasma ratio (D/P), peritoneal excretion, and clearance (PCl) of leptin were calculated. After 3 months of AADS administration (period II), leptinemia was transiently lower (9.8 +/- 6.2 ng/mL vs 17.1 +/- 14.2 ng/mL, p = 0.017), while D/P (0.51 +/- 0.44 vs 0.23 +/- 0.19, p = 0.012), peritoneal excretion (72.9 +/- 85.4 micrograms/day vs 37.2 +/- 32.3 micrograms/day, p = 0.015), and PCl (4.02 +/- 3.40 mL/min vs 1.75 +/- 1.32 mL/min, p = 0.008) of leptin were higher than measurements obtained at entry. Anorexia incidence and daily protein and energy intakes showed no significant changes during the study. Total body mass, body mass index, and plasma concentrations of total protein and of albumin increased significantly during AADS treatment. A significant positive relation of leptinemia to total fat mass was observed when AADS was not used (periods I and IV). We conclude that administration of AADS in CAPD patients causes a transient decrease in leptinemia and increases in peritoneal excretion and in PCl of leptin, as well as dissociation of the physiological relationship between serum leptin level and total fat mass. PMID- 11045253 TI - Long-term survival and its relationship to membrane transport status in peritoneal dialysis. AB - We evaluated peritoneal membrane transport status (high, high-average, low average, and low) of 35 patients on persistent peritoneal dialysis (PD) for 5-16 years. We noted that most of these patients (n = 21) had low-average transport by dialysate-to-plasma creatinine (D/PCr) ratio. PMID- 11045254 TI - Can the creatinine dialysate-to-plasma ratio from the peritoneal equilibration test be replaced by the sodium dialysate-to-plasma ratio and the sodium level in the dialysate? AB - The peritoneal equilibration test (PET) is a useful tool that categorizes peritoneal transport. However, the method has some inconveniences. Some authors suggest that measuring the sodium level in the dialysate (NaD) or the dialysate to-plasma ratio for sodium (D/PNa) can substitute for the PET. We applied a mathematical analysis [Fisher intraclass correlation coefficient (FICC)] to 43 PETs performed in 43 peritoneal dialysis patients (29 males, 14 females) with a mean age of 55.3 years (range: 28-85 years). Determinations of NaD, of sodium level in plasma (NaP), and of D/PNa at times 0, 30, 60, 120, and 240 minutes were added to the usual PET determinations. After using the NaD240 and the D/PNa240 values to calculate the cut-off values for the various peritoneal transport categories, we obtained a transport distribution very similar to that of the PET dialysate-to-plasma ratio for creatinine after 240 minutes (D/PCr240). At the same time, the FICC showed good (0.69) and excellent (0.77) correlation of NaD240 and D/PNa240 respectively with the D/PCr240. Therefore either of these two methods, which are cheaper and quicker than a PET, can be used to categorize peritoneal transport with a high degree of reliability. PMID- 11045255 TI - Necessity of correcting cancer antigen 125 appearance rates by body surface area. AB - Attention has been focused on cancer antigen 125 (CA125) levels in dialysate as an index for evaluating the severity of peritoneal mesothelial cell impairment. However, the method of evaluating CA125 levels in the dialysate has not been standardized. We determined that CA125 levels in the dialysate should be corrected by body surface area (BSA). Using 51 patients who had been undergoing peritoneal dialysis (PD), CA125 levels in the dialysate were measured to evaluate the appearance rate of CA125 (CA125AR). Because only 18 of the 51 patients (35%) had a BSA greater than 1.73 m2, CA125 levels in the dialysate might be underestimated in other patients when absolute values of CA125 are not corrected by BSA. When correlations between corrected CA125AR and various parameters were evaluated, corrected CA125AR positively correlated with age, but negatively correlated with serum creatinine and albumin levels. Moreover, no positive correlation was seen between corrected CA125AR and the duration of PD or the peritoneal permeability test. However, in 5 patients with ultrafiltration loss after prolonged PD, the values of CA125AR persistently or rapidly decreased before transfer to hemodialysis. Although CA125 can be used as an index of peritoneal deterioration, the absolute value of CA125 should be corrected by BSA. However, it is most important to evaluate changes in CA125 in individual patients. PMID- 11045256 TI - Ultrastructural changes of peritoneal lining cells in uremia. AB - Some thirty years ago, peritoneal dialysis (PD) became a respectable modality of renal replacement therapy. That is why peritoneal membrane attracted the interest of investigators. Uremia is followed by changes in the morphology of serous membranes (uremic serositis). Uremic effects on pleura and pericardia have been studied for a long time, but the peritoneum is affected as well. The aim of our study was to examine the morphology of the peritoneum in uremic patients before the start of PD and to compare the findings with those from examinations of peritoneum in healthy controls. We examined 12 uremic patients and 10 healthy controls (kidney donors). Biopsies were taken from parietal peritoneum. The samples were prepared in the standard way for study by transmission electron microscopy (TEM). Certain pathological changes--deformation of mesothelial cells, their detachment from the basement membrane, and unusual bulging of apical surface--were identified at the light microscopy level on semi-fine sections. Paracrystalline intracytoplasmic inclusions were seen in mesothelial cells only by TEM. We hypothesize that the inclusions were causing deformation of the mesothelial cells and detachment of those cells from the basement membrane. Sacculate dilatations of rough endoplasmic reticulum (RER) cisternae with partly destroyed membranes and few ribosomes were also seen, with and without densely osmiophilic filaments within cisternae. Although these changes are mentioned in the literature, the exact reason for their appearance remains unknown. PMID- 11045257 TI - Effect of dialysate composition on the apoptosis and proliferation of human peritoneal mesothelial cells and protein expression of Fas and c-Myc. AB - To investigate the effect of dialysate composition on apoptosis and proliferation of human peritoneal mesothelial cells (HPMCs) in vitro, HPMCs were cultured from human omental tissue. The cells were exposed for three days to culture medium containing: (1) 3.86% glucose, (2) 1.36% glucose, (3) 1.75 mmol/L Ca++, (4) 1.25 mmol/L Ca++, or (5) no additions (control). Apoptosis and proliferative cell nuclear antigen (PCNA) were measured by terminal deoxynucleotidyl transferase (TdT)-mediated in situ end-labelling and immunohistochemistry. Protein expression of Fas and of c-Myc were measured by flow cytometry. Apoptotic cells were significantly increased in the high glucose group compared to control (137 +/- 47 cells per field unit vs 63 +/- 30 cells per field unit, p < 0.01), but did not vary in the low glucose and low calcium groups versus control (76 +/- 25 cells per field unit and 77 +/- 31 cells per field unit, respectively, vs 63 +/- 30 cells per field unit, p > 0.05). Cells with positive expression of PCNA were increased in the high calcium group versus control (366 +/- 101 cells per field unit vs 186 +/- 76 cells per field unit, p < 0.01). However, the expression of PCNA was significantly more inhibited in the high glucose group than in the control group (158 +/- 52 cells per field unit vs 186 +/- 76 cells per field unit, p < 0.05). Expression of Fas was stimulated by high glucose (41% +/- 16% vs 25% +/- 8% in the control group, p < 0.05). High glucose and high calcium also up regulated the expression of c-Myc, but only the high glucose group showed significant difference from control (39% +/- 10% vs 24% +/- 8%, p < 0.05). Our data suggest that: (1) Peritoneal dialysate could induce apoptosis of HPMCs in vitro, the degree of apoptosis depending on concentration of glucose. (2) High calcium stimulates PCNA expression by HPMCs and high glucose inhibits expression of PCNA. (3) Genes associated with apoptosis (Fas, c-Myc) might play an important role in triggering apoptosis of HPMCs. PMID- 11045258 TI - Phospholipids in dialysate and the peritoneal surface layer. AB - Dialysate concentration of phospholipids has been used to monitor peritoneal membrane status. However, we recently found that the peritoneum has a surface layer in which phospholipids may be the main constituent. Therefore, in this study, we compared the phospholipids composition of peritoneal dialysate and of the peritoneal surface in rats. Eight male Sprague-Dawley rats were used in the study. Five rats received an intraperitoneal injection of 25 mL 4.25% glucose dialysis solution. After four hours, the rats were killed, and the dialysate was drained completely. Then 20 mL of Folch solution was infused into the peritoneal cavity for 30 seconds and drained completely. The other three rats received the Folch solution without dialysis. The effluent and Folch solution were then processed for phospholipids analysis using high performance thin-layer chromatography (HPTLC). The total phospholipids content was ten times higher in the surface layer than in the dialysate effluent. In the effluent, four clearly different components were seen: lysophosphatidylcholine (LPC), sphingomyelin (SM, 29%), phosphatidylcholine (PC, 66%), and phosphatidylinositol (PI, 4.5%). However, in the surface layer, as well as LPC, SM (20.6%), PHC (47%), and PI (6.3%), two additional components were seen, phosphatidylserine (PS, 17.1%) and phosphatidylethanolamine (PE, 8.9%). The quantity of phospholipids in the peritoneal surface of non dialyzed rats was similar to the total quantity of phospholipids (in effluent and in the peritoneal surface) of dialyzed rats. Our results suggest that: (1) a surface layer is present on the peritoneum; this layer could well be extracted by Folch solution; and, with appropriate incubation time, one can separate the surface layer without damaging the mesothelial cells; (2) the composition of phospholipids in the effluent is different from that in the peritoneal surface layer, which contains membrane phospholipids (PS and PE); (3) shielding from the peritoneal surface may be the main reason for the presence of phospholipids in the dialysate. PMID- 11045259 TI - Tranexamic acid: a "diuretic" for the peritoneal membrane? AB - The aim of the study was to use an animal model to investigate the mechanisms by which tranexamic acid (TNA) increases ultrafiltration (UF) volume. Thirty Sprague Dawley rats were studied in three groups (n = 10, each group). Group 1 was dialyzed with conventional dialysis solution, group 2 with conventional fluid plus low-dose TNA (300 mg/L), and group 3 with conventional fluid plus high-dose TNA (3000 mg/L). A PD catheter was inserted into each animal, and 25 mL of the designated dialysis fluid was injected twice daily for two weeks, starting seven days after surgery. Peritoneal transport properties (PTP) were measured by modified peritoneal equilibration test (PET), and transcapillary ultrafiltration (TCUF) and lymphatic absorption (LA) were determined by using dextran 70 weekly. Histological examinations of the peritoneum were performed at the end of the experiment. Drained dialysate volume (DDV) decreased in all groups; however, a lower reduction in DDV was seen in the TNA groups. PTP increased and TCUF decreased in all groups. LA increased in Group 1, but was unchanged in the other groups. No significant histological changes of the peritoneum were observed in any group. This study confirmed that, in rats, TNA maintained UF volume by reducing LA without serious adverse effects. We may safely use TNA as a "diuretic" for the peritoneal membrane. PMID- 11045260 TI - The role of peritoneal dialysis catheter configuration in preventing catheter tip migration. AB - Migration of the peritoneal dialysis (PD) catheter from the pelvis to the upper abdomen frequently results in peritoneal dialysis failure and removal of the catheter. Previous studies compared PD catheter survival in various catheter configurations. These studies included single-cuff and double-cuff, straight-end and curled-end catheters, and showed an incidence of catheter migration ranging from 5%-35% depending on the catheter type. Recent studies demonstrated that the double-cuff, Swan-neck, curled-end configuration is associated with a considerably lower incidence of migration. Most of these studies, however, had a small patient sample or no control group, or they compared nonequivalent catheters (for example, Swan-neck, curled-end versus straight, non-curled-end). Over a six-year period, we examined two similar double-cuff, curled-end catheters: Group I catheters had a straight segment between the two cuffs, and Group II catheters had a 60 degrees Swan-neck bend between the two cuffs (Quinton Instrument Co., Bothell, WA, U.S.A.). The two catheters were identical, except for the presence or absence of the Swan-neck bend. All catheters were placed by the closed laparoscopic technique. In the two groups of patients in whom the catheters were implanted, no statistically significant difference was observed in primary disease, age, sex, race, weight, prior abdominal surgery, or duration of dialysis before catheter migration. In group I, 33 of the 219 patients developed catheter migration (15%); in group II, 2 patients of 243 patients developed catheter migration (less than 1%, p = 0.002). In conclusion, the Swan-neck configuration presents an independent factor in preventing PD catheter migration. Review of previous studies and the data from our study, show that double-cuff, curled-end, Swan-neck PD catheters are superior to other catheters in regard to prevention of catheter migration and should be the catheter of choice in PD patients. PMID- 11045261 TI - Comparison of three chronic dialysis models. AB - The chronic peritoneal dialysis model is important for understanding the pathophysiology of peritoneal transport and for studying biocompatibility of peritoneal dialysis solutions. In this study, we compared three different chronic peritoneal dialysis models. A peritoneal catheter was placed in 23 male Sprague Dawley rats, 12 of which had an intact omentum (model 1) and 11 of which received an omentectomy (model 2). Seven other rats, without a catheter, received a daily intraperitoneal injection (model 3). Each rat received a daily infusion of 25 mL of 3.86% glucose dialysis solution either through the catheter (models 1 and 2) or through injection (model 3) for 4 weeks. Then, a 4-hour dwell study using 3.86% glucose solution with an intraperitoneal volume marker and frequent dialysate and blood sampling was performed in each rat. The intraperitoneal volume was significantly lower in all the dialysis groups as compared to a control group (n = 6) in which the rats had no chronic dialysate exposure. The peritoneal fluid absorption rate, as well as the direct lymphatic absorption rate, was significantly higher in the three dialysis groups as compared to the control group. In general, no significant differences were seen in any of the parameters among the three dialysis models. Owing to catheter obstruction, three rats in model 1 and four rats in model 2 were lost during dialysis. Histological examination showed no significant differences among the three dialysis groups. Our results suggest that omentectomy may not be necessary in the chronic peritoneal dialysis model when using dialysate infusion and no drainage. Based on the present study, we think that perhaps model 1 may be the method of choice to test new peritoneal dialysis solutions. However, owing to its simplicity, model 3 could also be used if great care is taken to avoid puncturing the intestine or injecting into the abdominal wall. PMID- 11045262 TI - Multivariable statistical methods for clinical studies: a gentle introduction. AB - Multiple regression models serve the same function as careful and thoughtful evaluation of tabular data. Two examples are given that show the value of holding one factor constant while examining the effect of another factor on an outcome of interest. PMID- 11045263 TI - Long-term survival on peritoneal dialysis in end-stage renal disease owing to diabetes. AB - Only a small number of peritoneal dialysis (PD) patients with diabetes have been followed for more than 5 years. Lack of extended follow-up for these patients is largely due to coexisting, far-advanced damage to target organs at initiation of dialysis, with progression of that damage during the course of dialysis; the presence of various comorbid conditions at the start of dialysis; and limits to long-term PD technique. Among renal replacement modalities, continuous ambulatory peritoneal dialysis (CAPD) has been favored by many clinicians for the treatment of diabetic patients owing to reasons inherent in the therapy. Reported survival rates of diabetic patients on CAPD vary from 17%-72% for the fourth year (mean value: 39%) and from 19%-63% for the fifth year (mean value: 35%). Diabetic patients have actuarial rates of patient survival and technique survival that are lower than those for non diabetic patients of comparable age on CAPD. Patient survival for diabetic patients undergoing PD is similar to that for diabetic patients on hemodialysis. Because the peritoneal membrane maintains its ability to adequately purify blood for a long time, CAPD remains a viable form of long term renal replacement therapy for diabetic patients with end-stage renal disease. PMID- 11045264 TI - Peritoneal dialysis in adult patients without end-stage renal disease. AB - End-stage renal disease remains the primary indication for the use of peritoneal dialysis. The therapy, however, has been used for the treatment of various other clinical conditions. Evidence has accumulated to support the use of peritoneal dialysis to maintain euvolemia, to improve functional status, and to reduce hospitalizations in patients with intractable chronic congestive heart failure. The use of peritoneal dialysis as a modality for core rewarming in patients with severe hypothermia has been established; in selected circumstances, it is probably the therapy of choice. The field of oncology has borrowed heavily from the technique of peritoneal dialysis for administering intraperitoneal chemotherapy; even though the therapy remains largely experimental today, it has great future potential. While efficacious in the treatment of acute, diuretic resistant volume overload in patients with congestive heart failure and in patients with severe, disabling psoriasis, the introduction of alternative methods of management have rendered the use of peritoneal dialysis obsolete. Finally, the role of peritoneal lavage in the management of patients with pancreatitis remains controversial and is no longer routinely used. PMID- 11045265 TI - Polyglucose dialysis solution influences serum iron parameters. AB - Owing to the lack of data dealing with the influence of polyglucose dialysis solution (PG-DS) on serum indicators of iron status, our study aimed at examining this problem in patients receiving PG-DS for the overnight exchange during treatment with continuous ambulatory peritoneal dialysis. We evaluated serum concentrations of iron, ferritin, and transferrin, total iron binding capacity (TIBC), and transferrin saturation (TSAT) at 1.6 +/- 0.8 months before introducing 7.5% PG-DS for an overnight 2 L exchange lasting about 10 hours (period I, n = 14), after 1.2 +/- 0.6 months of PG-DS administration (period II, n = 14), after 4.4 +/- 0.8 months of PG-DS administration (period III, n = 11), after 8.8 +/- 2.2 months of PG-DS administration (period IV, n = 9), and at 2.0 +/- 0.6 months after PG-DS discontinuation (period V, n = 11). Interference owing to PG-DS in laboratory determinations of serum iron parameters was excluded. Indices of nutritional status were also evaluated in all study periods. Significant differences in iron parameters were seen between periods I and III, or I and IV for transferrin (212 +/- 41 mg/dL vs 253 +/- 36 mg/dL), TIBC (304 +/- 40 micrograms/dL vs 338 +/- 31 micrograms/dL) and TSAT (34% +/- 15% vs 24% +/- 4%). After PG-DS withdrawal, these parameters all returned to pre-treatment values. Improvement in nutritional status was indicated by increases in total body mass (73.9 +/- 15.6 kg vs 77.4 +/- 13.8 kg), lean body mass (54.5 +/- 9.7 kg vs 56.9 +/- 8.5 kg), and serum total protein concentration (61.7 +/- 10.8 g/L vs 70.5 +/- 8.0 g/L). We conclude that serum transferrin concentration increases during PG-DS administration without enhanced iron binding to transferrin. An increase in transferrin level can be related to improved nutritional status. PMID- 11045266 TI - Infusion of total dose iron versus oral iron supplementation in ambulatory peritoneal dialysis patients: a prospective, cross-over trial. AB - The efficacy of intermittent, small doses of intravenous iron in hemodialysis patients is well established. But poor peripheral vascular access and frequency of therapy preclude acceptability of this method in peritoneal dialysis (PD) patients. Therefore, despite its marginal efficacy, oral iron remains the common method of iron supplementation in these patients. This prospective, cross-over trial compares infusion of total dose iron (ITDI) and oral iron supplementation in outpatient PD patients. Eleven stable CAPD patients with an hematocrit (Hct) of less than 33%, or a transferrin saturation (TSAT) of less than 30%, or both, were entered into the study. The study design included an oral phase [4 months, ferrous sulfate 325 mg (195 mg elemental iron), three times daily], a "wash-out" phase (1 month, no iron supplementation), and an ITDI phase [4 months, single infusion over 4 hours of 1 g iron dextran mixed in 1/2 normal saline]. Laboratory parameters were monitored monthly, and subcutaneous recombinant human erythropoietin (rHuEPO) doses were adjusted monthly to maintain a hematocrit above 33%. Patients with hyperparathyroidism, aluminum toxicity, and weekly Kt/V below 1.8 were excluded. Nine patients [8 Caucasians, 1 African American; causes of end-stage renal disease (ESRD): hypertension (4 cases), diabetes (3 cases), glomerulonephritis (1 case), and polycystic kidney disease (1 case); mean age: 43.3 +/- 2 years; mean weight: 73.0 +/- 4 kg; duration of ESRD: 28 +/- 13 months] completed the 9-month study. During the oral phase, TSAT rapidly decreased and a higher dose of rHuEPO failed to maintain Hct, a pattern sustained during the "wash-out" phase. During the ITDI phase, TSAT significantly increased and improvement in Hct resulted in a significant lowering of rHuEPO doses. The ITDI therapy was well tolerated. We conclude that ITDI is the preferred method of iron supplementation in outpatient PD patients. PMID- 11045267 TI - Serum level of vascular endothelial growth factor is influenced by erythropoietin treatment in peritoneal dialysis patients. (Grupo de Estudios Peritoneales de Madrid). AB - Some patients on long-term peritoneal dialysis (PD) develop a hyperpermeability state, owing to peritoneal neoangiogenesis. Vascular endothelial growth factor (VEGF), a potent mitogen for endothelial cells, has been implicated in most diseases characterized by microvascular neoformation. Erythropoietin (EPO) is able to induce endothelial proliferation in vitro. Our aim was to elucidate whether VEGF serum levels are influenced by EPO treatment, and whether VEGF serum level maintains a relationship with peritoneal transport data. We analyzed serum levels of VEGF in 35 PD patients (18 males, 17 females). Mean age was 58 years, with a mean time on PD of 98 +/- 75 months. Of the 35 patients, 19 were on automated peritoneal dialysis, and 16 were on continuous ambulatory peritoneal dialysis. Seven patients had diabetes. Peritoneal transport parameters were: urea mass transfer coefficient (MTC), 19.5 +/- 6.6 mL/min; creatinine MTC, 9.9 +/- 4.7 mL/min; net ultrafiltration, 491 +/- 166 mL per 4-hour dwell. Twenty seven patients were under therapy with recombinant human erythropoietin (rHuEPO). Mean serum VEGF levels were 347 +/- 203 pg/mL (range 66-857 pg/mL), with most patients in the normal range (60-700 pg/mL). VEGF levels did not correlate with age, sex, primary renal disease, diabetes, type of PD, time on PD, peritonitis, and cumulative glucose load. We found no correlation with urea MTC, creatinine MTC, ultrafiltration rate, or protein effluent levels. However, a significant negative correlation with residual renal function was seen (r = -0.39, p < 0.05). Patients treated with rHuEPO showed significantly higher serum levels of VEGF than non treated patients (375 +/- 220 pg/mL vs 251 +/- 75 pg/mL, p < 0.05), although they had similar residual renal function. We conclude that increased serum VEGF levels are associated with EPO treatment. Consequently, VEGF might have a role in the EPO effects found in PD patients. Whether both agents are related to peritoneal neoangiogenesis requires further research. PMID- 11045268 TI - Compliance with subcutaneous erythropoietin in peritoneal dialysis patients. AB - The use of erythropoietin in dialysis patients is associated with improved quality of life and may lead to improved outcomes. Peritoneal dialysis patients are generally taught to give subcutaneous erythropoietin at home. The purpose of this study was to determine compliance with erythropoietin use in peritoneal dialysis patients. Records of patient prescriptions, refills of erythropoietin, the person administering the injection, and average hematocrit were retrospectively examined. Demographic and comorbidity data were collected prospectively on all peritoneal dialysis patients. The use of more than 90% of the prescribed dose was considered compliant. Data were available for 55 patients. The mean follow-up time was 12 months. Thirty patients (55%) were noncompliant with erythropoietin. Compliant patients were older (65.4 years vs 50 years, p = 0.01), had higher comorbidity (p = 0.01), higher average hematocrit (34% vs 31.5%, p < 0.003) and were less likely to self-administer the injection. Sex, diabetes, and compliance with dialysis exchanges were not associated with erythropoietin compliance. Using logistic regression, the only significant variable was the person administering the erythropoietin (r = 0.46, p = 0.005). Noncompliance with subcutaneous erythropoietin is a significant problem. Compliance may improve if someone other than the patient is trained to administer it. PMID- 11045269 TI - Neoplasia in dialysis patients: pathophysiology, epidemiology, and screening. AB - Chronic renal failure can be considered an immunosuppressed state. One immunologic function that may be compromised is tumor surveillance. Studies have suggested an incidence of cancer that is greater in dialysis patients that in the age- and sex-matched non dialysis population. A drawback to many previous studies is "ascertainment bias," wherein closely monitored populations have a higher rate of cancer detection. However, allowing for systematic bias, a true increase in neoplasia appears to exist in dialysis patients. While many agencies have established protocols for cancer screening in the general population, little has been written about recommendations for cancer screening in dialysis patients. Moreover, the high mortality from cardiovascular disease may render cancer screening protocols redundant and cost-ineffective. This article offers recommendations that emphasize that risk profile of individual patients rather than a population-based approach to cancer screening. PMID- 11045270 TI - Impact of introduction of continuous ambulatory peritoneal dialysis on blood pressure: analysis of 24-hour ambulatory blood pressure. AB - Introduction of continuous ambulatory peritoneal dialysis (CAPD) induces a variety of hemodynamic changes in patients with end-stage renal failure. Among them, blood pressure (BP) control is likely to be the most important in determining long-term survival. The aim of our study was to use 24-hour ambulatory blood pressure monitoring (24 ABPM) to evaluate the previous and present BP status of patients on CAPD. We studied 18 non diabetic hypertensive patients who were introduced onto CAPD. At least 3-6 months before the introduction of CAPD, recordings of 24 ABPM were obtained. These recordings were then compared to others obtained 2-3 months after the introduction of CAPD. The average BP was higher before CAPD than after (160/91 +/- 10/5 mmHg vs 154/88 +/- 6/3 mmHg, p < 0.01). In addition, the doses and number of antihypertensive drugs used by the patients were reduced. Analysis of 24 ABPM revealed BP reduction predominantly in the daytime and less in the nighttime after introduction of CAPD. These results indicate that introduction of CAPD cannot reduce susceptibility of cardiovascular events, because nocturnal elevation of BP remains unchanged. We are therefore reminded that, after introduction of CAPD, casual BP measurements confined to the daytime may underestimate cardiovascular risk. PMID- 11045271 TI - Comparing an additional hour of cycler therapy to an additional midday exchange to achieve adequacy targets. AB - As CAPD patients lose residual renal function, adequate dialysis is frequently impossible to obtain unless the dialysis prescription is changed. For patients already on cycler therapy with a "wet" day, we compared the advantage of adding one hour on the cycler to adding an extra, midday exchange. To compare the two approaches, we used a commercial computer program to optimize solute clearance in 90 patients. Adding one hour of cycler therapy increases Kt/V and creatinine clearance (CrCl) by approximately 6.4%-8%. When a midday exchange is added, the increase in Kt/V and CrCl varies between 17.5% and 21.6%. We conclude that adding an extra, midday exchange always gives a better increase in weekly Kt/V and CrCl than that obtained by adding an extra hour of cycler therapy. PMID- 11045272 TI - Detection of subclinical abdominal hernia by peritoneal scintigraphy. AB - This study investigated the incidence of subclinical abdominal hernia in patients starting peritoneal dialysis (PD). From April 1995 to August 1999, every new patient without clinical evidence of abdominal leakage underwent peritoneal scintigraphy. A total of 59 patients were enrolled in the study. Imaging of the peritoneal cavity was performed by mixing 74 MBq (2 mCi) of 99 m technetium sulfur colloid with 2 L of 1.36% dextrose peritoneal dialysis solution. Sequential gamma camera static images were obtained at 0 minutes, 60 minutes, and after drainage. Ten abdominal hernias (2 diaphragmatic leaks, 8 inguinal hernias) were observed in ten patients (6 males, 4 females; mean age: 65.1 years). One patient with diaphragmatic leak recovered partial renal function and stopped continuous ambulatory peritoneal dialysis (CAPD); the other was switched to automated peritoneal dialysis (APD). Among the eight patients with inguinal hernia, six had no clinical manifestations within eight months of follow-up. Two patients became symptomatic at 15 months and 25 months respectively. They underwent surgical repair. In CAPD patients without obvious abdominal hernias, peritoneal scintigraphy at onset of dialysis discovered 17% positive cases. The technique of scintigraphy is safe, with a low radiation exposure. Surgical repair for maintenance on CAPD is not always necessary, and a change in the PD strategy may be useful. PMID- 11045273 TI - Combined peritoneal dialysis and hemodialysis therapy improves quality of life in end-stage renal disease patients. AB - The Dialysis Outcomes Quality Initiative (DOQI) guidelines recommend that, for patients on continuous ambulatory peritoneal dialysis (CAPD), a weekly creatinine clearance (WCC) of at least 60 L/1.73 m2 is needed for adequate dialysis. As residual renal function (RRF) declines, maintaining these target levels may become difficult. Over time, declines in ultrafiltration (UF) caused by increases in peritoneal permeability, in conjunction with decreases in RRF, may limit continuation of CAPD therapy. In an effort to achieve adequate solute clearance and ultrafiltration in several CAPD patients at our center who have declining RRF or poor UF, we use combined peritoneal dialysis and hemodialysis (PD + HD) as a therapeutic strategy when individualization of peritoneal dialysis is unsuccessful. At our center, PD + HD consists of five days of PD therapy followed by one HD session per week on Saturday. After the weekly HD session, patients are liberated from bag exchanges until Sunday evening. This PD + HD therapy was used in six cases where poor solute clearance and water retention were refractory to PD therapy alone. The combined therapy was well tolerated, and symptoms related to uremia improved in all six cases. Additionally, improvements in quality of life (QOL) were documented in all patients who were managed with the combined therapy. The improvements in QOL may have resulted from decreases in uremic symptomatology or freedom from bag exchanges. The PD + HD therapy can best be applied in the uremic PD patient without residual renal function whose peritoneal membrane is not deteriorated. The therapy allows for the continuation of PD without shifting to total HD in PD patients who continue to have uremic symptoms even after individualization of the PD prescription. Our patients readily accepted combined therapy, and we have noted excellent compliance with this therapy at our center. PMID- 11045274 TI - Polyglucose dialysis solution influences serum activity of amylase and of lipase differently. AB - Polyglucose dialysis solution (PG-DS) decreases serum amylase activity owing to interference in the analytical method. The interference can make it difficult to diagnose pancreatitis. Our aim was to check whether, during PG-DS administration, serum lipase activity changes simultaneously with serum amylase activity, and, if so, what the reason is for the detected change. Studies were started in 14 continuous ambulatory peritoneal dialysis (CAPD) patients in whom 7.5% PG-DS was applied for the overnight exchange. In addition to standard clinical and laboratory data, serum activity of lipase and of total amylase were evaluated at 1.6 +/- 0.8 months before PG-DS introduction (period I, n = 14), after 1.2 +/- 0.6 months of PG-DS administration (period II, n = 14), after 4.4 +/- 0.8 months of PG-DS administration (period III, n = 11), after 8.8 +/- 2.2 months of PG-DS administration (period IV, n = 9), and at 2.0 +/- 0.6 months after PG-DS discontinuation (period V, n = 11). The PG-DS was also added to serum from CAPD patients with known activity of amylase and of lipase. Immediately and 3 hours after PG-DS addition, a significant decrease in total amylase activity was seen; lipase activity was unchanged. In consecutive study periods, the results (median and range) were: for lipase activity--50 U/L (12-131 U/L), 59 U/L (25-160 U/L; p < 0.05 vs period I), 73 U/L (26-158 U/L; p < 0.05 vs period I), 66 U/L (30-203 U/L; p < 0.05 vs period I), and 44 U/L (15-112 U/L); for amylase activity--81 U/L (43-249 U/L), 14 U/L (5-82 U/L; p < 0.05 vs period I and period V), 15 U/L (5-192 U/L; p < 0.05 vs period I), 15 U/L (10-93 U/L; p < 0.05 vs period I and period V), and 118 U/L (4-221 U/L). An increase in serum lipase activity over the normal range (27-65 U/L) was not accompanied by clinical symptoms of pancreatic dysfunction, but rises were simultaneously shown in blood urea nitrogen, in serum level of creatinine and of total calcium, and in calcium phosphorus product. Our results confirm PG-DS influence on amylase determinations, exclude PG-DS interference in lipase measurements, and indicate that long-term PG-DS administration influences pancreatic exocrine function at a subclinical level. PMID- 11045275 TI - Does serum albumin at start of continuous ambulatory peritoneal dialysis (CAPD) or its drop during CAPD determine patient outcome? AB - The impact of serum albumin at start of continuous ambulatory peritoneal dialysis (SA1), serum albumin before death (SA2), and change in serum albumin during continuous ambulatory peritoneal dialysis (delta SA) were prospectively studied in 41 continuous ambulatory peritoneal dialysis (CAPD) patients with a follow-up of 19 +/- 11.6 months. For analysis, patients were divided into two groups at each measurement point: SA1 or SA2 > 3.0 g/dL or delta SA < 1.0 g/dL (group I), and SA1 or SA2 < 3.0 g/dL or delta SA > 1.0 g/dL (group II). On log rank test, the mortality rate was significantly higher in group II patients in relation to SA1 (p < or = 0.0001), SA2 (p = 0.0002), and delta SA (p = 0.001). On univariate Cox proportional hazard regression analysis, SA1 (p = 0.0001), SA2 (p = 0.0001), delta SA (p = 0.002), and episodes of peritonitis (p = 0.0001) were significant. On multivariate analysis, SA2 (p = 0.003) was significantly related to patient mortality. SA2 (r = 0.8; p = 0.0001), but not delta SA, was related to SA1. We conclude that SA2 is the best predictor of patient mortality on CAPD. SA2 is strongly related to SA1. Thus protein restriction in the pre-dialysis stage should be advised cautiously to avoid consequent hypoalbuminemia. PMID- 11045276 TI - Subcutaneously tunnelled peritoneal dialysis catheters with delayed externalization: long-term follow-up. AB - We performed a retrospective analysis of our institution's experience with the technique of delayed externalization of subcutaneously tunnelled peritoneal dialysis catheters. From 1993 to 1999, 49 catheters were implanted in 37 patients. Median age of the patients was 43.6 years; 70% were female; 32% had diabetes. Most of the catheters were midline, single-cuff, curled Quinton catheters without a swan neck. One patient underwent transplantation prior to catheter externalization. One catheter leaked prior to externalization and was removed. The remaining catheters were externalized a median of 40 days (range: 18 319 days) post implantation. At externalization, two leaks and one omental obstruction occurred, causing primary catheter failure. Total days of catheter follow-up were 17,895. One-year and two-year catheter survival rates were 70% and 40% respectively. Catheter failure occurred owing to infection in 7 cases and to mechanical complications in 10 cases. The rate of exit-site infection was 1 per 9.9 patient-months, and of peritonitis, 1 per 16.2 patient-months. Including primary failures, mechanical complications were 12 hernias, 6 leaks, and 4 instances of malposition. We conclude that delayed externalization of single-cuff catheters without a swan neck is associated with increased mechanical and infectious complications. These findings may warrant a change to a double-cuff catheter with a swan neck. PMID- 11045277 TI - Prevalence and causes of cough in chronic dialysis patients: a comparison between hemodialysis and peritoneal dialysis patients. AB - Peritoneal dialysis (PD) and hemodialysis (HD) are both common forms of dialysis for patients with end-stage renal disease. A few case reports have suggested that cough is associated with PD. From 1991 to 1998, 17 patients being treated with PD at the Toronto Western Hospital demonstrated persistent cough severe enough for referral to a respirologist. Causes of cough, often more than one cause per patient, included asthma, post-nasal drip, gastroesophageal reflux disease (GERD), chronic obstructive pulmonary disease, congestive heart failure, allergic rhinitis, pleural effusion, and respiratory infection. The aim of this cross sectional study was to establish the prevalence of cough among PD patients, to determine if PD patients more commonly have a dry persistent cough than do HD patients, and, if the latter case is true, the possible reasons for it. A detailed survey of 92 PD patients and 91 HD patients was conducted in 1998 and 1999 at the University Health Network. Survey questions inquired about patient respiratory symptoms since onset of dialysis. Charts were reviewed to obtain information on use of medications possibly relevant to cough. In the PD and HD groups, 52% and 23% were females (p = 0.001), and the mean ages were 59.1 and 60.1 years, respectively. Angiotensin converting enzyme (ACE) inhibitors had been taken by 65% (PD) and 55% (HD) of patients, and beta-blocking medications by 43% (PD) and 51% (HD). Since initiation of dialysis--mean 2.7 years (PD) and 3.7 years (HD)--22% of PD patients reported persistent cough versus 7% of HD patients (p = 0.003). Although no significant association was seen between cough and self reported heartburn in HD patients (p = 0.439), a significant association between cough and self-reported heartburn was seen in PD patients: 67% of PD patients with persistent cough reported heartburn versus 29% of those without cough (p = 0.008). The findings suggest that GERD and associated cough are more common in PD patients than in HD patients, perhaps owing to increases in intra-abdominal pressure from the peritoneal dialysate. PMID- 11045278 TI - The law of unintended consequences in action: increase in incidence of hypokalemia with improved adequacy of dialysis. AB - In 1996, we raised our peritoneal dialysis (PD) dose to meet new DOQI adequacy targets. Concurrently, we noted an increase in the frequency of K+ levels below 3.5 mEq/L. A continuous quality improvement (CQI) project was initiated to quantify the impact of increasing dialysis dose on the prevalence of hypokalemia in our unit. Measurements of serum K+, blood urea nitrogen (BUN), creatinine, residual renal function, and the number and type of clinical interventions required to maintain eukalemia were abstracted from the charts of 62 patients enrolled in our program for more than 6 months and having more than two adequacy data points. In the seven consecutive 6-month periods from January 1996 to June 1999, dialysis dose progressively increased while median serum K+ decreased, and the percentage of patients requiring either diet counselling or K+ supplementation rose from 9% to 42%. We conclude that the increased clearance of K+ that occurs with increasing dialysis dose may lead to significant hypokalemia in a large proportion of PD patients dialyzed to DOQI adequacy targets. Maintenance of eukalemia in this population often requires increased K+ intake and or oral supplementation. Further studies are needed to ascertain whether the prevalence of hypokalemia is sufficient to warrant routine addition of K+ to PD dialysis solutions. PMID- 11045279 TI - Body mass index in patients with amputations on peritoneal dialysis: error of uncorrected estimates and proposed correction. AB - "Weight-height" indices including percent of ideal weight (%IW) and body mass index (BMI) are used to estimate degree of obesity in populations and are predictors of survival in dialysis patients. Amputation affects the relationship between weight and height independently of the degree of obesity. Corrections of both %IW and BMI for amputation have been published, but a National (U.S.) computer nutrition program used in the authors' institution uses only the correction for %IW. This study had two parts: (1) To test whether the weight height cut-off values for weight deficit (%IW 90%, BMI 20 kg/m2) and obesity (%IW 120%, BMI 30 kg/m2) are compatible, we performed linear regression of BMI on %IW in peritoneal dialysis (PD) patients without amputations. In 349 men, BMI = 0.834 + 0.226 (%IW), r = 0.979. From this regression, the 95% confidence interval (CI) of BMI is 19.2-23.1 kg/m2 if %IW is 90%, and 26.1-29.9 kg/m2 if %IW is 120%. In 260 women, BMI = 2.194 + 0.184 (%IW), r = 0.974. From this regression, the 95% CI of BMI is 15.7-21.8 kg/m2 if %IW is 90%, and 21.3-27.3 kg/m2 if %IW is 120%. (2) To identify the direction and magnitude of the error of uncorrected BMI (BMIu) in dialysis patients with amputations, we analyzed weight-height indices in two groups of men by the computer nutrition program, which corrects %IW, but not BMI for amputation, and by the corrected BMI (BMIc) formula. In group A (amputation without height loss, n = 11), %IW = 110.2% +/- 16.9%, BMIu = 23.6 +/- 2.7 kg/m2, BMIc = 26.4 +/- 3.8 kg/m2 (p < 0.001, BMIc vs BMIu), and 5 of the 11 BMIu values fell below the 95% confidence band of the regression of BMI on %IW in patients without amputations. In group B (amputation with loss of height, n = 6), %IW = 92.7% +/- 19.9%, BMIu = 33.9 +/- 10.7 kg/m2, BMIc = 22.1 +/- 4.4 kg/m2 (p < 0.005, BMIc vs BMIu), and 5 of the 6 BMIu values fell above the 95% confidence band of the regression of BMI on %IW in patients without amputations. CONCLUSIONS: (1) The weight deficit cut-offs for %IW and BMI are compatible in non amputated men and women. (2) The obesity cut-offs for %IW and BMI are compatible in non amputated men, but not in non amputated women. (3) Amputation without height loss decreases BMIu, while amputation with height loss increases, in general, BMIu. (4) BMI should be corrected in PD patients with amputations. PMID- 11045280 TI - The relationship between peritoneal clearance and patient outcome in peritoneal dialysis patients. AB - Peritoneal clearance has not yet been proven to predict clinical outcome in peritoneal dialysis patients independent of residual renal function. Studies looking at the relationship between clearance and outcome have not been adequately powered to detect an effect. Pending better studies, clearance targets have been established, and higher clearances are being delivered. This article reviews existing literature on the topic and discusses the problems associated with carrying out a randomized controlled trial to address the issue. PMID- 11045281 TI - Peritoneal equilibration test in Indian patients on continuous ambulatory peritoneal dialysis: does it affect patient outcome? AB - The variations in peritoneal equilibration test (PET) characteristics in various ethnic groups have been reported. Data are scarce regarding the pattern of membrane characteristics in Indian patients. The factors affecting PET and the PET, in turn, affecting patient outcome are controversial issues. We prospectively analyzed 41 patients to evaluate: (1) the pattern of PET characteristics in Indian patients; (2) the factors affecting the PET; (3) the effect of membrane characteristics on patient outcome. The mean period of follow up was 17.1 +/- 9.3 months. The PET results revealed 21 high transporters (51%), 13 high-average transporters (32%), 6 low-average transporters (15%), and 1 low transporter (2%). The distribution of the various PET categories in patients below the age of 60 years and those 60 years or above was not significant (p = 0.70). The sex distribution (p = 0.94) and prevalence of diabetes (p = 0.62) were not significantly different in various PET categories. On regression analysis, PET values were not affected by the age of patients (beta = 0.80, p = 0.61). Patient survival among high and high-average transporters was significantly less compared with low and low-average transporters (p = 0.01). We conclude that Indian patients on continuous ambulatory peritoneal dialysis (CAPD) have a higher proportion of high and high-average transporters. The pattern of membrane kinetics cannot be explained by differences in patient characteristics and diabetic status. Patients with high PET values have poorer patient survival on CAPD. PMID- 11045282 TI - Coagulation and fibrinolysis factors in dialysis patients with and without ischemic heart disease. AB - Cardiovascular disease is the leading cause of death in patients with end-stage renal disease. In addition, abnormalities of coagulation and fibrinolysis have been reported in patients with uremia. However, whether these hemostatic abnormalities lead to cardiovascular disease in dialysis patients is currently unknown. Therefore, we investigated the association of hemostatic factors with ischemic heart disease (IHD) in patients on peritoneal dialysis and hemodialysis. The study patients comprised 30 continuous ambulatory peritoneal dialysis patients and 18 hemodialysis patients. Twenty healthy subjects served as controls. We evaluated each subject's hemostatic factors, including factor VII, factor XII, thrombin-antithrombin III complex (TAT), fibrinogen, plasmin antiplasmin complex (PIC), plasminogen activator inhibitor (PAI-1), and D-dimer. In dialysis patients, IHD was diagnosed by documented myocardial infarction or positive result on coronary angiogram or by positive thallium myocardial scintigraphy. Factor VII, fibrinogen, PIC, and D-dimer levels were significantly higher in the two dialysis groups than in controls. All hemostatic variables were similar between the two dialysis groups. Subject age (p = 0.005), PIC (p = 0.005), and D-dimer level (p = 0.003) were significantly higher in patients with IHD than in patients without IHD in the dialysis groups. Multiple logistic regression analysis showed that only patient age and D-dimer levels were independent predictors of IHD. Adjusted odds ratio for IHD was 1.06 for each 10 ng/mL increase of D-dimer (p = 0.06). In CAPD patients, only D-dimer was independently associated with IHD (odds ratio: 1.06, p = 0.03). We conclude that multiple hemostatic abnormalities are present in dialysis patients and that elevated D-dimer levels are independently associated with prevalent IHD. PMID- 11045283 TI - Incidence of sleep pattern disturbance in a peritoneal dialysis sample. AB - Our experience and the research literature suggest that sleep pattern disturbance (SPD) is a problem among dialysis patients. The purpose of this study was to describe the scope of sleep problems among willing patients on peritoneal dialysis at a mid-size university teaching hospital. To examine SPD, this descriptive study used a sleep diary that patients completed each morning for a week. Patients were asked to describe sleep latency, perceived difficulty falling asleep, number of arousals, use of sedatives, whether they awoke feeling rested, sleep efficiency, and factors preventing or inducing sleep. The sample consisted of 22 respondents whose average age was 60.5 years. With respect to the variables included in the sleep diary, respondents reported: 82% sleep latency of 40 minutes or less; of 154 nights studied, difficulty falling asleep on 41 (27%) of the nights; 82% experience of 15 or fewer sleep arousals per week (that is, approximately 2 arousals per night); 64% no use of sedatives during the week; 55% experience of feeling rested on awakening 5 or more mornings per week; and 55% experience of sleep efficiency above 80%. Of the factors reported to interfere with sleep, treatment-related factors such as alarms and other machine-related problems were, by far, the most predominant, reported by 82% of respondents. These findings compare very favorably with reports in the literature noting the incidence of SPD among dialysis patients to be as high as 73%. Information regarding factors that are barriers or facilitators to sleep have formed the basis of some practice changes within our program to address this distressing problem. PMID- 11045284 TI - Adaptation of the Fresenius PD+ Cycler for a hearing-impaired patient. AB - Continuous cycling peritoneal dialysis (CCPD) uses a cycler to perform dialysis exchanges and requires the patient to respond to an audible alarm signifying an interruption in the therapy. Consequently, an unassisted hearing-impaired patient could not use the system. By converting the standard alarm to a vibrating signal, the cycler was successfully adapted to accommodate the special needs of our hearing-impaired patient. The items required for the modification were the Sonic Alert Wake Up Alarm (Model SA-WA300: Sonic Alert, Troy, MI, U.S.A.) and the Sonic Alert Super Shaker Bed Vibrator (Model SA-SS120V: Sonic Alert). The patient can place the vibrator under either the pillow or the mattress. When the cycler alarm is activated, vibration wakens the patient. The equipment was purchased from Harris Communications (Eden Prairie, MN, U.S.A.) through a referral by the Easter Seal Society. Three days were needed to complete training compared to an average of one or two days for patients previously trained for continuous ambulatory peritoneal dialysis (CAPD). The patient remained on cycler therapy for approximately four months when the unrelated development of an abdominal hernia required termination of peritoneal dialysis and subsequent transfer to hemodialysis. In conclusion, a modified cycler can provide a safe and efficient renal replacement therapy option for a hearing-impaired patient. PMID- 11045285 TI - Placement of peritoneal dialysis catheter by percutaneous method with the Veress needle. AB - In our center, a total of 451 peritoneal dialysis catheters were implanted in 424 uremic patients from 1986 to 1999. All catheters were implanted by the percutaneous method of Di Paolo. From 1986 to May 1997 (period A), we inserted 389 catheters using the Weston-Roberts catheter armed with a metallic sharp mandrel for the first access to the peritoneal cavity. After May 1997 (period B), we implanted 62 peritoneal dialysis catheters, replacing the metallic sharp mandrel with the Veress needle to arm the Weston-Roberts catheter. The aim of this retrospective study was to determine the incidence of early complications (within 2 weeks) and to compare the incidence between the two groups (period A versus period B). The results suggest that the incidence of perforation of abdominal organs is lower, but not significantly so, when the Veress needle is used. PMID- 11045286 TI - Peritoneal dialysis is the therapy of choice for end-stage renal disease patients with hereditary clotting disorders. AB - Chronic renal failure is an unusual complication of hereditary clotting disorders (HCDs), but this situation could change in the near future. The modality of dialysis for end-stage renal disease (ESRD) in patients with an HCD is a difficult choice. Hemodialysis (HD) may be considered, but intensive treatment with coagulation factors is required for vascular access execution and for each HD procedure. Peritoneal dialysis (PD) has been infrequently proposed. However, PD requires coagulation replacement therapy only during peritoneal catheter placement. The aim of this paper is to describe our experience of three patients with ESRD and HCD, successfully treated with chronic PD in the medium term. Case 1 was a 58-year-old man with moderate hemophilia A, type 2 diabetes mellitus, and hepatitis C virus (HCV) infection. His ESRD was secondary to glomerulonephritis. A double-cuff peritoneal catheter was surgically placed with pre-emptive factor VIII administration. He began treatment with continuous ambulatory peritoneal dialysis (CAPD). An inguinal hernia was repaired without complications. After eleven months of follow-up, no hemorrhage episodes have been observed and clinical outcome is optimal. Case 2 was a 46-year-old man with severe hemophilia A, type 2 diabetes mellitus, and HCV and human immunodeficiency virus (HIV) infections. He developed a diabetic nephropathy that required renal replacement therapy. A permanent silicone catheter was inserted in the left internal jugular vein, and the patient started HD treatment. Later on, PD therapy was proposed. A peritoneal catheter was implanted with simultaneous factor VIII infusion. Minimal bleeding was observed at the subcutaneous tunnel over the following 48 hours. The patient started PD treatment without complications, and two months later, remaining asymptomatic, transferred to another center. Case 3 was a 41-year-old woman diagnosed with von Willebrand disease type 2A, HCV infection, and polycystic kidney disease, who presented with ESRD. An internal arteriovenous fistula was performed under coagulation factor cover. During a fistulography, and despite coagulation factor substitutive treatment, the patient showed an important hematoma. Afterwards, PD was considered. A peritoneal catheter was implanted under coagulation factor cover. The postoperative course was uncomplicated, and the patient started CAPD treatment. During follow up, she suffered two hemoperitoneum episodes that were resolved with cold dialysate. After nine months, she uneventfully continued on PD. In conclusion, PD is the therapy of choice for patients with hereditary clotting disorders and ESRD requiring dialysis. Peritoneal dialysis therapy avoids many of the complications related to HD therapy. PMID- 11045287 TI - Ultrafiltration with icodextrins in continuous ambulatory peritoneal dialysis and automated peritoneal dialysis. AB - Icodextrins (Icos) produce constant linear ultrafiltration (UF). This effect allows Icos to replace glucose during long dwells in continuous ambulatory peritoneal dialysis [CAPD (nighttime)] and automated peritoneal dialysis [APD (daytime)]. However, the effectiveness of Icos in producing UF (IcoUF) is limited by lymphatic reabsorption, whose extent depends partly on posture and physical activity. This paper aims to assess whether the difference in posture and physical activity between daytime dwells in APD and nighttime dwells in CAPD affects IcoUF. Patients undergoing first treatment were retrospectively examined. Ten patients were on CAPD [4 males, 6 females; average age, 73.0 +/- 13.4 years; body surface area (BSA), 1.63 +/- 0.21 m2; total volume per day, 5.6 +/- 1.9 L], and ten were on APD (7 males, 3 females; average age, 67.7 +/- 9.8; BSA, 1.75 +/- 0.22 m2; total volume per night, 10.5 +/- 0.9 L). Ultrafiltration was assessed for seven consecutive days preceding a peritoneal equilibration test (PET) and collection of diuresis. In both groups, 3 patients had no diuresis, and the difference between CAPD and APD was not significant (625 +/- 762 mL vs 780 +/- 878 mL). Moreover, no significant difference was seen in 4-hour dialysate-to plasma creatinine (D/P) between CAPD (0.65 +/- 0.12) and APD (0.64 +/- 0.05). Dwell times with Icos were shorter in CAPD than in APD (11.5 +/- 1.8 hours vs 14.8 +/- 0.5 hours, p < 0.0005), but the fill volume was not significantly different (1760 +/- 286 mL vs 1790 +/- 249 mL). Water excretion owing to diuresis and dialysis [total water excretion (TWE): 1619 +/- 497 mL CAPD vs 1762 +/- 736 mL APD] and dialytic UF (363 +/- 443 mL CAPD vs 748 +/- 479 mL APD), which is not linked to Icos, were not significantly different between the two groups. The IcoUF and the percentage of IcoUF to TWE were significantly higher in CAPD compared to APD [631 +/- 253 mL (44% +/- 27%) vs 234 +/- 215 mL (19% +/- 19%), p < 0.001 (p < 0.05)]. In conclusion, an upright posture and physical activity seem to produce less IcoUF in APD despite the longer dwell. These factors could, indeed, produce greater intraperitoneal pressure, resulting in increased lymphatic reabsorption during a daytime dwell. PMID- 11045288 TI - What happens after conversion of treatment to continuous ambulatory peritoneal dialysis from hemodialysis? AB - A prospective study was planned to follow the clinical and laboratory data of hemodialysis (HD) patients after change of treatment to continuous ambulatory peritoneal dialysis (CAPD). Patients who had been on the HD program for more than 6 months were selected and followed for at least 6 months under CAPD treatment. Measured parameters included hemoglobin, ferritin, C-reactive protein (CRP), calcium, phosphorus, and intact parathyroid hormone (iPTH) levels; lipid profile; total protein and albumin; body mass index and triceps skin fold thickness; echocardiographic findings; and medications administered. We followed 34 patients (12 males, 22 females; mean age: 43.5 +/- 14.5 years; mean HD duration: 36.6 +/- 24.76 months) for a mean period of 19.8 +/- 11.9 months after change of treatment to CAPD. We saw a significant increase in mean hemoglobin, cholesterol, triglyceride, high-density lipoprotein (HDL), lipoprotein (a) [Lp(a)], phosphorus, and iPTH levels. We observed a decrease in erythropoietin dose, mean ferritin levels, systolic blood pressure (139.4 +/- 22.8 mmHg vs 114.4 +/- 21.0 mmHg, p = 0.001), diastolic blood pressure (85.7 +/- 12.6 mmHg vs 73.5 +/- 17.6 mmHg, p = 0.002), percentage of left ventricular hypertrophy, systolic and diastolic dysfunction, and the number of hypertensive drugs received. A significant improvement in the nutritional status of the patients (total protein, body mass index and triceps skin fold thickness) was also seen. In conclusion, CAPD treatment has a short-term outcome superior to that of HD in terms of better nutritional status and better control of hypertension and anemia. PMID- 11045289 TI - The impact of the earthquake in northwestern Turkey on the continuous ambulatory peritoneal dialysis patients who were living in the earthquake zone. AB - In August 1999, an earthquake of magnitude 7.8 on the Richter scale hit northwestern Turkey. The epicenter was in Izmit, an industrial town about 60 km from Istanbul. This paper presents data about the fate of CAPD patients who were living in that region at the time of the earthquake. A total of 42 continuous ambulatory peritoneal dialysis (CAPD) patients (14 females, 28 males; 37 adult patients, 5 pediatric patients) were permanent residents of the earthquake region. They were followed in the CAPD units of Marmara University Hospital (n = 6), Gata Military Hospital (n = 2), and Goztepe SSK Hospital (n = 10, including the 5 pediatric patients) in Istanbul, and in Uludag University Hospital in Bursa (n = 6) and Kocaeli University Hospital in Izmit (n = 18). Two CAPD patients, together with their families, died under the rubble in the city of Golcuk. One CAPD nurse from Kocaeli University Hospital in Izmit also died a victim of the earthquake. One patient who lived in Golcuk was under the rubble for 3 hours; she was rescued with no crush injuries and was able to continue with CAPD 24 hours after her rescue. Eight patients reported that their homes were completely destroyed during the earthquake, while nine patients reported serious damage to their houses. Ten patients had to move to other towns to live with relatives because their homes were no longer suitable for habitation, and twelve patients had to stay permanently in tents provided by the Red Cross. All of the patients were able to continue their CAPD therapy and had no interruption in the supply of their CAPD solutions. Four patients on continuous cycling peritoneal dialysis (CCPD) therapy continued to use their HomeChoice machines (Baxter Healthcare Corporation, Deerfield, IL, U.S.A.) even while living in a tent. CAPD patients from the Kocaeli University Hospital had to be temporarily referred to other CAPD centers in Istanbul and Bursa because the Kocaeli University Hospital was seriously damaged in the earthquake. We expect that these major changes in quality of life circumstances will have an important impact on the morbidity of these patients, especially in regard to the rate of peritonitis and the adequacy of dialysis. PMID- 11045290 TI - Education as a clinical tool for self-dialysis. AB - Therapeutic compliance and patient education are presently considered crucial parts of end-stage renal disease (ESRD) therapy. In the center where Italian home and self-care dialysis treatment started, an education program was designed as multi-step pathway--following patients from chronic renal failure to dialysis and eventual graft--employing lessons, booklets, and books. Each step was validated in various subsets of patients. Lessons involved two hours of informal discussion on the main aspects of ESRD and renal replacement therapy (RRT); booklets were created from tape recordings of the lessons. Patient participation was good, with 28 of 33 patients on charge in the center for 6 months or more taking part in more than one lesson in 1999. In 16 of 16 patients who answered a questionnaire after two lessons, expressed opinion was "good" to "fair." All asked for further material. With regard to books, 500 copies of the book What does dialysis mean? were given out in the region; this book was validated in 22 patients on peritoneal dialysis (PD) and 18 on hemodialysis (HD). It helped patients to accept dialysis in 65% of cases and to comprehend it in 90%. Four thousand copies of the book Stories, containing 18 interviews on transplantation, were printed, and this book was validated in 21 patients on self-care and 35 on hospital dialysis (potential candidates for graft). Of 56 patients, 53 asked for further material; 19 changed their initial opinion (10 choose transplantation, despite initial skepticism; 9 put off transplantation, despite initial acceptance). On a local scale, the program led 12 of 18 new patients, who followed at least part of the program, to choose self-dialysis (PD, home, and self-care dialysis). PMID- 11045291 TI - Telemedicine system for home automated peritoneal dialysis. AB - We have developed a new telemedicine system to monitor elderly and handicapped patients that use an automated peritoneal dialysis (APD) system to perform continuous ambulatory peritoneal dialysis (CAPD) at home. The system has two parts: (1) a data collection and transport system, and (2) an interview system ("View Send" system). The former is assembled from the APD cycler, an automated blood-pressure recorder, and a scale. The latter is a combination of a digital camera, television, and set-top box (a kind of computer). The APD system contains recording and data transport junctions, a monitoring device, and a data tabulation function. All data collected by the APD system are sent directly in real time to the physician's office. Patients can easily use this telemedicine system to contact the medical staff at the Saitama Medical School CAPD center and to consult concerning their condition. Furthermore, the staff can directly change the CAPD schedule (dose and duration). Seven patients, including some who are older than 90 years or who are handicapped, have been using this telemedicine system for between 1 and 6 months (average: 3 months). From our recent experience, our tentative conclusions are that (1) elderly and handicapped patients benefit from this system by being able to maintain CAPD without major problems and accidents; (2) other problems are mainly due to simple mistakes such as accidentally pushing the on and off switches; (3) quality of life is improved for the patients. PMID- 11045292 TI - Simultaneous removal and reinsertion of Tenckhoff catheters for the treatment of refractory exit-site infection. AB - Exit-site infection (ESI) refractory to medical therapy is an important complication of continuous ambulatory peritoneal dialysis (CAPD). Between July 1994 and June 1998, 28 patients in our hospital underwent simultaneous removal and reinsertion of Tenckhoff catheters for the treatment of refractory ESI. The ESI was caused by Pseudomonas aeruginosa in 22 patients (78%), methicillin resistant Staphylococcus aureus in 5 patients (18%), and diphtheroid bacilli in 1 patient (4%). The patients had received antibiotic treatment for a mean duration of 11.6 +/- 5.8 weeks (range: 3-28 weeks) before their operations. During each operation, the old Tenckhoff catheter was removed and a new catheter was inserted in the opposite side of the abdomen. CAPD was resumed after two weeks of intermittent peritoneal dialysis. All patients received intravenous antibiotic cover for seven days after the operation. Early post-operative complications were uncommon. At one year after the operation, 22 patients (78%) were free of ESI. Six patients (22%) had a recurrence of ESI 21.3 +/- 6 weeks (range: 16-32 weeks) after their operation. All new infections were treated successfully with antibiotics. Seven patients (25%) had one episode of peritonitis each, all of which resolved with intraperitoneal antibiotic treatment. We conclude that simultaneous removal and reinsertion of Tenckhoff catheters is a safe and effective method for the treatment of refractory ESI. The procedure alleviates the need for temporary hemodialysis and allows an early return to CAPD. PMID- 11045293 TI - Preoperative vancomycin prophylaxis for newly placed peritoneal dialysis catheters prevents postoperative peritonitis. AB - The role of vancomycin and other antibiotics in treatment of acute peritonitis in peritoneal dialysis patients is well established. However, the role of preoperative vancomycin or cephalosporins in preventing early infection in newly placed peritoneal dialysis catheters remains controversial. We performed a prospective randomized study to examine the role of vancomycin or cefazolin prophylaxis in decreasing the incidence of postoperative peritonitis. Over 8-year period, 265 patients undergoing 305 permanent peritoneal catheter placement procedures were randomized into three groups. Group I (103 procedures) received a single intravenous (i.v.) dose of 1000 mg vancomycin 12 hours before the peritoneal catheter placement procedure. Group II (102 procedures) received a single i.v. dose of 1000 mg of Ancef (cefazolin) 3 hours before the procedure. Group III (100 procedures) received no antibiotics preoperatively for a least one week before the procedure. Patients were monitored for peritonitis during the following 14 days. Peritonitis developed in 1 patient (1%) in Group I (vancomycin group) compared to 12 patients (12%) in Group III (control group), p = 0.002, and in 9 patients (9%) in Group II (cefazolin group) compared to Group III, p = 0.68. We conclude that the use of preoperative single-dose i.v. vancomycin prophylaxis for permanent peritoneal dialysis catheter placement reduces the risk of postoperative peritonitis. Cefazolin did not achieve a statistically significant difference from the control group and may not provide adequate prophylaxis. PMID- 11045294 TI - Pathophysiology and morphological clinical correlation in experimental and peritoneal dialysis-induced peritoneal sclerosis. AB - The definition of peritoneal sclerosis encompasses a vast range of peritoneal alterations induced by peritoneal dialysis. The morphology, clinical correlations and pathophysiology of, and experimental animal models for, this condition show such striking differences from case to case as to suggest the existence of two nosological entities: simple sclerosis and sclerosing peritonitis. PMID- 11045295 TI - In vitro study of the efficacy of a two-way connection with disinfectant in the prevention of peritonitis. AB - A new connection system for continuous ambulatory peritoneal dialysis (CAPD) has been established, and its efficacy in preventing microbial contamination of the peritoneal cavity has been tested in vitro. The system consists of a Y-shaped channel formed in the bottom of a Plexiglas cup. The Luer-lock shaped ends of the Y-shaped channel are designed to host the connectors from the drainage bag, the catheter transfer set, and the bag of fresh dialysate. Because the connectors from the catheter transfer set and the fresh bag are located at the inner surface of the cup bed, and because the cup is filled with disinfectant during the entire exchange procedure, all at-risk steps are continuously protected by disinfectant (that is, removal of the caps from the connectors, connection and disconnection, replacement of the caps). Still, because the patient could inadvertently extract and contaminate one of the two connectors (although such a possibility is unlikely), the disinfecting efficacy of the system was tested in vitro. Despite contamination with various micro-organisms at the highest possible concentrations, all tests showed negative bacterial growth, thus confirming the absolute efficacy of the system in preventing exogenous transluminal peritonitis. PMID- 11045296 TI - CD40 ligand expression on macrophages during peritonitis in continuous ambulatory peritoneal dialysis patients. AB - CD40-CD40 ligand (CD40L) interaction plays an important role in macrophage/monocyte-mediated inflammatory processes by up-regulating cytokine production by macrophages/monocytes and by preventing macrophage apoptosis at the inflammation sites. The present study investigated the possible regulation of CD40L expression in peritonitis during continuous ambulatory peritoneal dialysis (CAPD). We used fluorescence-activated cell sorter (FACS) analysis to detect CD40L expression on macrophages obtained from peritoneal dialysate. Our results showed that CD40L expression on macrophages was significantly increased in a peritonitis group (4.62 +/- 6.54) as compared to a control group (0.76 +/- 0.30, p < 0.01). The CD40L-positive cells were also significantly increased during peritonitis (97.86% +/- 1.67% in the peritonitis group as compared to 73.10% +/- 26.94% in the control group, p < 0.05). After successful treatment, the expression of CD40L was significantly reduced (3.66 +/- 1.12 vs 1.05 +/- 0.02, p < 0.05). We conclude that functionally expressed CD40L on macrophages may take part in acute inflammatory response during peritonitis in CAPD and may play an important role in the local defense against infection in the peritoneal cavity. PMID- 11045297 TI - Natural anti-alpha-galactosyl antibodies in patients undergoing peritoneal dialysis and hemodialysis. AB - Anti-alpha-galactosyl antibodies (anti-Gal) seemingly mediate rejection of pig organs transplanted into humans and Old World monkeys. These natural antibodies appear to be produced by a subpopulation of B cells residing in the peritoneum. Therefore, peritoneal dialysis (PD) may have an impact on anti-Gal levels. In this cross-sectional study, blood anti-Gal levels were quantified by ELISA in 17 patients undergoing PD and 18 patients undergoing hemodialysis (HD), and the results were compared with those from a control group of 30 healthy blood donors. The effects of the mode of dialysis therapy and other epidemiologic variables on anti-Gal levels were also evaluated. The PD and HD patients were comparable with regards to age, sex, percentage having diabetes, time on dialysis, distribution of blood groups, and total serum levels of immunoglobulins A (IgA), G (IgG), and M (IgM). In patients and controls, IgG anti-Gal levels were not significantly different, but IgM anti-Gal levels were significantly lower in both PD and HD patients compared with controls. A nonsignificant trend toward lower IgM anti-Gal levels in PD patients compared with HD patients was also observed. IgG anti-Gal levels correlated positively with time on dialysis in the HD patients, but not in the PD patients. IgG anti-Gal levels also were found to be markedly elevated in three patients with chronic liver disease, but no other scrutinized variable appeared to influence IgG or IgM anti-Gal levels. PMID- 11045298 TI - Total creatinine appearance as indicator of risk of infectious complication in peritoneal dialysis. AB - Peritonitis and exit-site infections are the main causes of complications in peritoneal dialysis. Death due to infectious complication is also one of the major causes of drop-out. The underlying cause of infection may include malnutrition. Total creatinine appearance (TCA) may reflect overall nutritional status. We determined TCA from the daily dialysate, urine, and estimated gut creatinine of patients and normalized it to actual body weight (nTCA). We examined the relationship between nTCA and the incidence of infection, and between nTCA and infection-related survival. The study included 323 adult patients in a single dialysis center. The mean nTCA of all patients was 19.73 +/- 4.75 mg/kg/day. The patients with an nTCA below 1 standard deviation from the mean (nTCA < 14.98 mg/kg/day) had a significantly higher peritonitis and exit site infection rate (p < 0.01) and a higher chance of drop-out owing to infection related complications (p < 0.0001). Our study concluded that the adult patient with malnutrition (nTCA < 14.98 mg/kg/day) has higher risk of infection. PMID- 11045299 TI - The spectrum of peritoneal fibrosing syndromes in peritoneal dialysis. AB - A variable degree of diffuse peritoneal fibrosis has been documented in all patients who have been on long-term peritoneal dialysis. Peritoneal dialysis induced diffuse peritoneal fibrosis varies from opacification and "tanning" of the peritoneum, which may have only a moderate detrimental effect on peritoneal transport kinetics, to a progressive, sclerosing encapsulating peritonitis (SEP), which may lead to cessation of peritoneal dialysis and to death. Fewer than 1% of peritoneal dialysis patients develop overt SEP as manifested by combinations of intestinal obstruction, weight loss, and ultrafiltration failure. The diagnosis of SEP depends on a combination of laparotomy and radiological features in suspected cases and consequently the true incidence of SEP is most likely underestimated. Several predisposing, interrelated risk factors for both peritoneal fibrosis and sclerosing encapsulating peritonitis have been identified: prolonged duration of peritoneal dialysis, history of severe or recurrent episodes of peritonitis, and higher exposure to hypertonic glucose based dialysis solutions. Nevertheless, the etiology of SEP is unknown and several causal factors may simultaneously or sequentially initiate and maintain a low-grade serositis that leads to uncontrolled fibroneogenesis. The high mortality rate of SEP has emphasized the need to develop preventive strategies. These strategies include early peritoneal catheter removal to avoid refractory peritonitis, the development of more biocompatible dialysis solutions, restriction of the use of hypertonic glucose-based dialysis solutions during and after episodes of peritonitis, and, perhaps, limiting the duration of peritoneal dialysis in at-risk patients. This approach was followed in a Japanese unit where a subgroup of all patients who had been on peritoneal dialysis for more than 5 years and who had poor ultrafiltration and peritoneal calcification on computed tomography (CT) scan were shown to have peritoneal sclerosis on peritoneal biopsy and were therefore electively transferred to hemodialysis. This acquired spectrum of peritoneal fibrosing syndromes leads to long-term complications in peritoneal dialysis, whereas localized fibrous adhesions secondary to prior abdominal surgery may prevent the successful initiation of peritoneal dialysis. PMID- 11045300 TI - Peritonitis and antibiotic therapy in patients on cycler peritoneal dialysis--an update. AB - The increased use of automated peritoneal dialysis (APD) and the inherent differences between continuous ambulatory peritoneal dialysis (CAPD) and APD have generated interest in the treatment of peritonitis in cycler patients. This review considers variations in the incidence of peritonitis and in its microbiological spectrum among CAPD and APD patients, and discusses the potential causes for these variations, with emphasis on recent literature. Flow-pattern variances between CAPD and APD demand special considerations in the diagnosis of peritonitis. Multiple alternatives for the management of peritonitis in APD are discussed in light of recent clinical experiences and pharmacokinetic considerations. PMID- 11045301 TI - Role of intraperitoneal urokinase in acute peritonitis and prevention of catheter loss in peritoneal dialysis patients. AB - Some studies have demonstrated the efficacy and safety of intraperitoneal (i.p.) urokinase in the resolution of recurrent or relapsing peritonitis, while others have not. Most studies were small, and they varied in methodology. Furthermore, the role of i.p. urokinase in shortening the duration of peritonitis or in preventing recurrence after initial peritonitis has not been examined. In addition, no previous studies have examined the role of i.p. urokinase in preventing, after first infection, catheter loss due to unresolving (resistant) peritonitis. Over a period of 3 years, we prospectively randomized into two groups all peritoneal dialysis (PD) patients who developed a first episode of peritonitis. Group I (n = 40) received i.p. urokinase on the first day of diagnosis (5000 IU intraluminally in the peritoneal catheter and left for 4 hours before next exchange). Group II (n = 40) received no urokinase. The duration of peritonitis was assessed by daily PD fluid white blood cell (WBC) count. Indications for catheter removal were: persistent peritonitis after four days from initiation of antibiotic therapy, or peritonitis with multiple organisms, suggesting bowel perforation. No statistically significant difference was seen between the two groups in regard to primary cause of end-stage renal disease (ESRD), age, sex, race, weight, type of dialysis [continuous ambulatory peritoneal dialysis (CAPD), automated peritoneal dialysis (APD), continuous cycling peritoneal dialysis (CCPD)], or duration of dialysis prior to first peritonitis. No statistically significant difference was seen between the two groups in the duration of peritonitis or in the severity of symptoms and signs of peritonitis. Neither was any difference seen in the peritonitis recurrence or relapse rate (10% in the urokinase group vs 7.5% in the control group). Nine patients lost their catheters (3 in the urokinase group: 1 Pseudomonas aeruginosa and 2 Candida tropicalis; 6 in the control group: 1 Klebsiella pneumonia, 1 enterococcus, 2 Pseudomonas aeruginosa, and 2 Candida tropicalis). The difference in the rate of catheter loss between the two groups was not statistically significant; it appeared to relate to the type of organism rather than to the response to urokinase. In conclusion, i.p. urokinase plays no significant role in shortening the course of peritonitis or in preventing recurrence or loss of the PD catheter. Loss of PD catheters in patients having their first peritonitis appears to be related primarily to the type of organism causing the infection. PMID- 11045302 TI - High prevalence of selective immunoglobulin A deficiency in peritoneal dialysis patients. AB - We encountered two hemodialysis (HD) patients with recurrent infections and complete immunoglobulin A deficiency (IgAD). To survey the possibility of a similar occurrence in other populations, we conducted the present study. We used nephelometry to examine the levels of immunoglobulins G (IgG), A (IgA), and M (IgM) in 42 continuous ambulatory peritoneal dialysis (CAPD) patients, 246 HD patients, 56 chronic renal failure (CRF) patients, and 250 normal adults. Four CAPD patients (9.5%) and five HD patients (2.0%) were found to be completely IgA deficient (IgA < 6.67 mg/dL). Peritoneal dialysis patients therefore had a significantly higher prevalence of IgAD compared with HD and CRF patients. The underlying diseases leading to dialysis therapy in the IgAD patients varied. Their dialysis durations also varied. The occurrence rate of peritonitis in CAPD patients with IgAD was no higher than in patients without IgAD. The clinical significance of IgAD was focused on mucosal immunity, but the exact prevalence of infection was difficult to define. However, these patients' medical records did suggest more frequent respiratory tract infections and cellulitis events than did the records of patients without IgAD. Two PD patients with IgAD died of pneumonia. Immunodiffusion and indirect ELISA methods were used to detect the presence of auto-antibodies, successfully identifying them in three patients. Further research is needed to study other mechanisms of IgAD. PMID- 11045303 TI - The influence of seasonal factors on the incidence of peritonitis in continuous ambulatory peritoneal dialysis in the temperate zone. AB - Many factors contribute to the development of continuous ambulatory peritoneal dialysis (CAPD) peritonitis. The role of climate in CAPD peritonitis has received relatively little attention. We retrospectively analyzed the incidence of CAPD peritonitis according to temperature and relative humidity. Data from 80 patients were examined. The monthly mean temperature and relative humidity during study period varied between -3.4 degrees C and 25.4 degrees C and between 61% and 81%, respectively. In 1123 patient-months, 53 cases of peritonitis occurred. The occurrence of peritonitis paralleled temperature and relative humidity, being the highest (0.180 episodes/patient-month) in July (mean temperature, 24.6 degrees C; relative humidity, 81%) and lowest (0.013 episodes/patient-month) in November (mean temperature, 6.6 degrees C; relative humidity, 66%). Significant correlations were seen between the monthly frequency of CAPD peritonitis and temperature (r = 0.53, p < 0.05) and relative humidity (r = 0.59, p < 0.05). The incidence was higher in the warm season (months with a mean temperature > or = 15 degrees C, that is, May-September) than in the cold season (months with a mean temperature < 15 degrees C, that is, October-April), at 0.074 episodes/patient month versus 0.024 episodes/patient-month, p < 0.05. We also found a tendency for gram-negative peritonitis to occur uniformly throughout the year, but for gram positive peritonitis to increase during hot and humid months, especially the rainy month of July. Gram-positive organisms caused 50% of peritonitis from March to August, but just 17.7% from September to February. Gram-negative organisms caused 7.3% and 29.4% of peritonitis during the same periods (p < 0.05). The results indicate a clear seasonal change in the rate of CAPD peritonitis and in the causative micro-organisms. The observation that CAPD peritonitis increases in the season of high temperature and high humidity suggests the influence of climate on CAPD peritonitis in the temperate zone. PMID- 11045304 TI - Clonotypes of Staphylococcus aureus isolated from continuous ambulatory peritoneal dialysis patients: what is the vector between nares and infection site? AB - Staphylococcus aureus is frequently isolated from patients with infections related to continuous ambulatory peritoneal dialysis (CAPD). In many cases, the organism is also isolated simultaneously from the anterior nares. To clarify the transmission trail of S. aureus, we used DNA analysis to identify clonotypes of clinical strains. The nares and exit sites of 32 CAPD patients were swabbed, and PD fluid samples were taken for pathogen culture. Genome DNA of S. aureus was digested with restriction enzyme Sma I for pulsed-field gel electrophoresis. We also asked the patients how they usually performed the PD procedure. S. aureus was isolated from 4 patients, including 3 who hosted two strains isolated separately from different sites. The DNA patterns of the strains isolated from these latter 3 patients were identical. However, the clonotypes from all 4 patients were different. Most of the patients did not wash their hands and wear masks while exchanging PD bags and caring for their exit sites. After the patients were disinfected and re-educated in proper procedures, S. aureus was not detected in any of them. These data suggest that no outbreak occurred in our hospital and that the vectors of endogenous infection were the patients themselves, probably their hands. A bacteriological study presents an efficient opportunity to re-educate patients in PD procedure. PMID- 11045305 TI - Surgical treatment for sclerosing encapsulating peritonitis. AB - Sclerosing encapsulating peritonitis (SEP) is recognized as a serious complication of continuous ambulatory peritoneal dialysis (CAPD). To date, in our hospital, 12 cases of SEP have been successfully treated by active intervention. The development of SEP was observed in these patients after removal of a peritoneal catheter. SEP was relieved by steroid administration in 3 of these patients, and by total parenteral nutrition (TPN) performed after exploratory laparotomy in 1 patient. In the remaining 8 patients, SEP was relieved by total intestinal enterolysis. In patients who underwent total intestinal enterolysis, the severity of encapsulation and adhesion varied. White, rigid encapsulation was observed in 4 patients who had been treated by peritoneal dialysis (PD) for less than 10 years. Seemingly normal serosae were observed under the capsules, and total intestinal enterolysis was easily performed in these patients. In the patient who underwent renal transplantation, more severe intestinal adhesion was observed, although the duration of PD was limited to 70 months and the intestinal serosae were seemingly normal. These findings were considered specific to SEP developing after immunosuppressant administration. In 3 patients who had undergone PD for more than 10 years, degeneration of the visceral peritoneum was observed, together with an ill-defined boundary between the capsules and the serosae. Therefore, total enterolysis was performed in these patients, including a wide area of the muscular layer. Furthermore, calcification was observed in several regions, where the capsules were severely adherent to the parietal peritoneum. The post-operative course for all 8 patients was satisfactory, and these patients finally returned to their previous social activities. We conclude that when SEP symptoms are not improved by steroid administration or TPN, active total intestinal enterolysis should be performed. However, it is absolutely important to avoid inducing anastomosis or impairing the intestine. PMID- 11045306 TI - The effectiveness of mupirocin preventing Staphylococcus aureus in catheter related infections in peritoneal dialysis. AB - The objective of this study was to evaluate the effectiveness of mupirocin on Staphylococcus aureus with regard to peritoneal dialysis (PD)-catheter exit-site infections (ESI), tunnel infections (TI), and peritonitis episodes (PE). The study was performed on 42 continuous ambulatory peritoneal dialysis (CAPD) patients (group I) treated from April 1998 to July 1999. These patients were instructed to apply mupirocin daily at the catheter exit site as part of their exit-site care. The control was the same group's historical infection data. Results were also recorded for a second group of 16 patients (group II) with newly implanted PD catheters were also instructed to apply mupirocin at the exit site daily. During the control period (before daily mupirocin application), group I recorded 16 episodes of ESI (0.30 episodes per patient-year), 6 episodes of TI (0.11 episodes per patient-year), 15 episodes of PE (0.28 episodes per patient year), and one case of catheter removal (0.019 episodes per patient-year) owing to S. aureus exit-site infection coexisting with peritonitis. The rate of S. aureus exit-site infection during this period was 0.11 episodes per patient-year; of S. aureus tunnel infection, 0.057 episodes per patient-year; and of S. aureus peritonitis, 0.076 episodes per patient-year. During the mupirocin period, infections and peritonitis owing to S. aureus dramatically decreased (p < 0.01 and p < 0.001 respectively). The rate of S. aureus exit-site infection was 0.02 episodes per patient-year, with no S. aureus tunnel infections, and no catheter removals owing to S. aureus peritonitis. Similarly, in group II, no episodes were recorded of any ESI, TI, or PE owing to S. aureus, although 4 episodes of ESI (0.37 episodes per patient-year, 2 with other gram-positive bacteria, and 2 with gram-negative bacteria) and 8 PEs (0.75 episodes per patient-year) were seen. We conclude that mupirocin application provides excellent prophylaxis for catheter related infections owing to S. aureus, and that reduction of these infections may improve the long-term survival of patients on CAPD. PMID- 11045307 TI - Hypertonicity of dialysis fluid suppresses intraperitoneal inflammation. AB - We studied acute and chronic intraperitoneal inflammation during dialysis performed in rats injected with phosphate-buffered saline alone (PBS) or PBS supplemented with glucose (Glu) or with mannitol (Man). In acute experiments, the result of a first dialysis with PBS in every rat (dialysis I) was compared with a second dialysis performed 24 hours later (dialysis II) using a different dialysis fluid: either PBS with 3.86 g/dL Man or PBS with 3.86 g/dL Glu. In rats exposed to hypertonic dialysis solutions (both Glu and Man), inflammatory reaction was decreased (cell count: p < 0.05; nitric oxide secretion: p < 0.05; protein in dialysate: p < 0.05). In control animals treated only with PBS (dialysis I and dialysis II), the inflammatory reactions during dialysis I and dialysis II were comparable. In chronic experiments, rats were dialyzed with the tested fluids for four weeks. Weekly, dialysate samples were taken and analyzed. At the end of the study, cell counts and protein losses were higher in the PBS-treated rats than in the other groups (cell count: p < 0.05, Glu vs PBS, and p < 0.05, Man vs PBS; protein in dialysate: p < 0.001, Glu vs PBS, and p < 0.01, Man vs PBS). We conclude that hypertonicity of the dialysis fluid inhibits the inflammatory reaction in the peritoneal cavity during peritoneal dialysis. PMID- 11045308 TI - A prospective multicenter comparison of peritonitis in peritoneal dialysis patients aged above and below 65 years. Levante PD Multicenter Group. AB - Various authors have found peritonitis rates in older peritoneal dialysis (PD) patients to be higher, similar, or even lower when compared to rates in younger populations. We prospectively analyzed all episodes of peritonitis registered in the 381 patients (219 males, 162 females; mean age, 55.5 +/- 17.0 years) who were treated with PD during four years (1993-1996) in our multicenter group. Patients were distributed into two groups. Group A included 138 patients aged 65 years or over (mean age: 72.3 +/- 5.1 years); group B included 243 patients aged below 65 years (mean age: 46.0 +/- 13.6 years). No differences were seen in general PD characteristics. Normalized protein catabolic rate (nPCR) was higher in younger patients (p = 0.001). Patients in group A experienced more peritonitis (135 episodes; 1 episode per 16.7 patient-months) than patients in group B (198 episodes; 1 episode per 21.7 patient-months; p = 0.01). Although no differences were seen in the general characteristics of the peritonitis episodes, gram negative peritonitis and peritonitis not achieving a cure were more commonly secondary to enteric bacteria in group A (p = 0.03). We conclude that PD patients aged 65 years or over are at higher risk of peritonitis. Also, in this age group, gram-negative peritonitis and peritonitis with any evolution except cure are more likely to be due to enteric bacteria. PMID- 11045309 TI - Outcome and clinical implications of a surveillance and treatment program for Staphylococcus aureus nasal carriage in peritoneal dialysis patients. AB - Staphylococcus aureus nasal carriage (SANC) is a risk factor for development of S. aureus dialysis-related infections. Reported here are results of a SANC surveillance and treatment program employed by our dialysis unit over a two-year period. Surveillance nasal cultures were performed at 3-month intervals in 129 peritoneal dialysis patients. Those with SANC applied mupirocin ointment intranasally 3 times daily for 5 consecutive days for 3 consecutive months. Treatment was repeated only when subsequent cultures showed SANC. Infection and catheter loss rates were compared to 63 historical controls, and between SANC and non SANC patients of the study group. Patients who were initially non carriers showed increasing probability for acquiring SANC throughout the study period. Following treatment, the probability for recurrence of SANC was 26%, 41%, 58%, and 62% at 1, 3, 6, and 12 months. The rates of S. aureus exit-site or tunnel infection (p = 0.36), peritonitis (p = 0.0002), and catheter loss (p = 0.01) were lower in the study group as compared to controls. Despite treatment, SANC patients demonstrated a twofold increase in exit-site/tunnel infection rate (p = 0.03) and a threefold increase in catheter loss rate (p = 0.1) as compared to non SANC patients. The high rate of SANC recurrence and the long interval between surveillance cultures may explain the failure of the current protocol to completely eliminate the risk for S. aureus infections. The results support a change in the treatment plan to that of continuing the monthly mupirocin regimen indefinitely once SANC has been identified. PMID- 11045310 TI - Prevalence of microbial colonization in removed peritoneal catheters: a prospective study. AB - We conducted a prospective bacteriologic study of 89 peritoneal catheters removed from 77 peritoneal dialysis patients. Reasons for catheter removal included severe peritonitis (n = 36, Group A), persistent exit-site infection (n = 29, Group B), and dormant, seemingly uninfected catheters (n = 22, Group C). We studied the external cuff (EC) and internal cuff (IC) as well as the catheter tip. In Group A, microbial growth was seen in 86.1% of ECs, 66.7% of ICs, and 67.6% of tips. In cases of positive isolation, concordance was 91.7% IC versus EC, 84.2% IC versus tip, and 80.0% EC versus tip. The peritonitis agent was recovered from 61.1% of ECs, 50.0% of ICs, and 55.6% of tips. In Group B, colonization was seen in 72.4% of ECs, 44.8% of ICs, and 31.0% of tips. When an isolation was obtained from both EC and IC, concordance was 81.8%. The exit-site infection agent was recovered from 69% of ECs and 24% of ICs. In Group C, microbial growth was observed in 77.3% of ECs, 45.5% of ICs, and 31.8% of tips. Gram-positive bacteria predominated, with the same bacteria colonizing EC and IC in 66.7% of cases. In conclusion, removed peritoneal catheters present a high prevalence of extensive microbial colonization, even in the absence of overt infection. PMID- 11045312 TI - Spectrum of complications related to secondary hyperparathyroidism in a peritoneal dialysis patient. AB - The index patient is a 23-year-old female with end-stage renal disease (ESRD) secondary to chemotherapeutic agents. Continuous cycling peritoneal dialysis (CCPD) has been the renal replacement therapy for the past 5 years since a failed cadaveric renal transplant. Past medical history was significant for diabetes mellitus, hypertension, anemia, bilateral subclavian vein thrombosis with superior vena cava syndrome, secondary hyperparathyroidism, leukemia (at age 8), and hyperlipidemia. On presentation, soft tissue nodules were noted in the anterolateral surfaces of the legs. After 3 months of continued low-calcium dialysate CCPD, calcitriol, and oral phosphate binders, a 2 x 3 cm nodule was noted on the posterior aspect of the thorax at the scapula. The only complaint at this time was shoulder pain at the acromioclavicular joint. Radiological examination revealed a 3 x 4 cm soft tissue opacity in the superior segment of the left lower lobe laterally. Despite a prior subtotal parathyroidectomy, phosphate binders, and calcitriol, the parathyroid hormone levels continued to increase, with development of tumoral calcinosis, worsening renal osteodystrophy, and calciphylaxis. Computed tomography examination revealed extensive soft tissue calcification consistent with tumoral calcinosis. An ulcerative lesion (1 cm) developed on the lateral aspect of the upper thigh owing to warfarin necrosis versus calciphylaxis. At this time, the phosphate binder was changed from calcium acetate to sevelamer hydrochloride. Aggressive wound treatment and aggressive calcium and phosphate control added to the treatment regimen has resulted in healing of the single ulcer and a decrease in the size of the tumoral lesions. In conclusion, early recognition and aggressive treatment of calciphylaxis can result in reduced morbidity and mortality from calciphylaxis in ESRD patients. PMID- 11045311 TI - Use of bolus intraperitoneal aminoglycosides for treating peritonitis in end stage renal disease patients receiving continuous ambulatory peritoneal dialysis and continuous cycling peritoneal dialysis. AB - Peritonitis is the most common complication of peritoneal dialysis (PD) and accounts for the most drop-out from PD among patients with end-stage renal disease (ESRD). Antibiotic protocols to treat peritonitis recommend initial treatment for both gram-positive and gram-negative infections, pending culture results. Current literature also suggests comparable therapeutic success using bolus (single-dose) parenteral aminoglycosides to treat systemic gram-negative infections compared to conventional divided-dose aminoglycosides. The single bolus dose antibiotic regimen has the potential for reduced nephrotoxicity and ototoxicity. We therefore hypothesized that intermittent bolus dose intraperitoneal (i.p.) aminoglycosides may be superior to conventional continuous dose i.p. aminoglycosides in treating dialysis-associated peritonitis, and have fewer side effects. Six patients with clinical peritonitis were treated with single, bolus-dose, i.p. gentamicin or tobramycin (5 mg/kg ideal body weight), range 250-440 mg (mean: 355 +/- 68.25 mg) at the start of therapy. No patient grew gram-negative organisms; aminoglycosides were therefore not repeated. Three patients used four continuous ambulatory peritoneal dialysis (CAPD) exchanges per day; three patients used six nightly continuous cycling peritoneal dialysis (CCPD) exchanges with a daytime dwell. Mean PD aminoglycoside clearance was 6.75 +/- 2.27 mm3/min; mean urinary aminoglycoside clearance was 5.75 +/- 1.14 mm3/min. Mean blood aminoglycoside elimination t1/2 was 29.27 +/- 3.55 hours, with a mean blood level of 3.18 +/- 1.45 micrograms/mL 72 hours after initial therapy, and 1.52 +/- 0.81 micrograms/mL 96 hours after initial therapy. Peritoneal equilibration test (PET) scores before and after aminoglycoside administration (performed a mean of 4.6 months apart) were 0.672 +/- 0.097 and 0.705 +/- 0.092 respectively (p = 0.321). Comparative audiograms using pure-air tone conduction with frequencies from 250-12,000 Hz were done within 24 hours of aminoglycosides and again when therapy was complete (mean: 17 days). No significant changes were seen. While efficacy of bolus versus conventional-use aminoglycosides could not be definitely established, the kinetics of bolus aminoglycosides suggests that therapeutic blood levels persist for 72-96 hours and that the risk for oto/vestibular toxicity is negligible. We conclude that use of bolus i.p. aminoglycosides is safe, achieves therapeutic blood levels for extended intervals, demonstrates no clinical oto/vestibular toxicity, is cost effective, and is a convenient strategy for patients and nursing staff. PMID- 11045313 TI - Protein nutrition status of adult patients starting chronic ambulatory peritoneal dialysis. AB - Malnutrition is an important determinant of outcome in patients on dialysis. Its cause is multifactorial and its detection is important when patients embark on dialysis. In this study, we used various indices to assess the prevalence of malnutrition as reflected in protein intake, anthropometric effect, and serum protein level. To avoid the effect that dialysis has on malnutrition, we studied 37 patients who had just started continuous ambulatory peritoneal dialysis (CAPD). Of these new patients, 42% had a residual urea clearance below 1.0 mL/min. In 51% of the patients, serum albumin level was below 33 g/L; in 14%, body mass index was below 19; and in 39%, dietary protein intake was below 0.8 g per kilogram of body weight per day. A total of 76% of our patients had at least one of these three indices. In 8% of the patients, lean body mass was below 60%. We conclude that protein malnutrition is a significant problem in our patients starting CAPD. Dialysis-dependent factors were not implicated, as the patients were studied at the start of dialysis. Low intake of protein was a major problem and may have contributed significantly to malnutrition. The low residual urea clearance implied that dialysis was started relatively late and may have contributed to the low protein intake. PMID- 11045314 TI - Influence of adynamic bone disease on responsiveness to recombinant human erythropoietin in peritoneal dialysis patients. AB - Adynamic bone disease (ABD) has an increasing prevalence in the dialysis population, more so in peritoneal dialysis patients. Anemia in patients with high turnover bone disease and high intact parathyroid hormone (iPTH) tends to be resistant to recombinant human erythropoietin (rHuEPO). The same problem may occur in patients with ABD; however, data are scarce. This study evaluates the effectiveness of rHuEPO in 32 chronic peritoneal dialysis patients, 9 with iPTH levels below 100 pg/mL for more than 6 months (group A, with ABD) and 23 with iPTH levels above 100 pg/mL (group B, without ABD). In group A and group B respectively, the dosage of rHuEPO was 141.8 +/- 59 U/kg/week and 144.8 +/- 77 U/kg/week, and hematocrit was 33.2% +/- 4.3% and 31.7% +/- 4.5% (p > 0.05). Iron indices, nutritional parameters, and bone indices were similar, except that group A had lower alkaline phosphatase and serum ferritin levels. The data suggest that patients with ABD may not be resistant to rHuEPO, but may even have a slightly better hematocrit at a similar rHuEPO dosage. Further studies in a larger number of patients are needed to confirm these findings. PMID- 11045315 TI - Glucocorticoid restores the deterioration of water transport in the peritoneum through increment in aquaporin. AB - This study was carried out to investigate the role of aquaporin (AQP) in the peritoneum undergoing continuous ambulatory peritoneal dialysis (CAPD). Furthermore, we examined the effects of treatment with prednisolone (PSL) in a rat model of peritoneal sclerosis. We modelled peritoneal sclerosis by using dialysis solution with the addition of 0.1% chlorhexidine gluconate (CHG) for 10 days. Twenty male Wistar Kyoto (WKY) rats were divided into four groups and dialyzed with various solutions: (1) saline (NS group, n = 5); (2) 10% glucose (TZ group, n = 5); (3) 0.1% CHG (CHG group, n = 5); and (4) 0.1% CHG plus PSL (CHG + PSL group, n = 5). Expression of mRNA of AQPs (AQP-1-AQP-4) was studied by semi-quantitative reverse-transcription polymerase chain reaction (RT-PCR). Expression of AQP-4 was also measured by Western blot analysis. Ultrafiltration volume and peritoneal function were measured by the peritoneal equilibration test. In the TZ group, expression of AQP-1 and AQP-4 were significantly enhanced, in parallel with an increment in ultrafiltration volume. On the other hand, in the CHG group, expression of AQP-1 and AQP-4 were significantly suppressed, and ultrafiltration volume was lost. The use of PSL with CHG completely restored the expression of AQP-1 and AQP-4, and peritoneal function improved. No expression of AQP-2 and AQP-3 was seen in the peritoneum. Our results suggest that AQP-1 and AQP-4 may be important factors in water transport in patients undergoing CAPD. PSL might be an effective treatment to prevent the progress of peritoneal sclerosis in patients undergoing CAPD. PMID- 11045316 TI - Pulse oral versus pulse intraperitoneal calcitriol: a comparison of efficacy in the treatment of hyperparathyroidism and renal osteodystrophy in peritoneal dialysis patients. AB - Controversy exists among various studies in regard to the efficacy of oral (p.o.) versus parenteral calcitriol. Some studies suggest that intravenous (i.v.) calcitriol is superior to p.o. calcitriol for treating renal osteodystrophy in hemodialysis patients; others suggest that these routes of administration are equivalent. To our knowledge, no large, prospective, randomized study compares intraperitoneal (i.p.) to p.o. calcitriol in adult peritoneal dialysis patients. We conducted a prospective randomized study in 76 patients (38 on i.p. calcitriol and 38 on p.o. calcitriol), whom we followed for 48 months. Of the 76 patients, 34 (18 in the i.p. group and 16 in the p.o. group) completed the 48-month study period. Calcitriol dosing was similar in both groups (3-6 micrograms per week in three divided doses). Dose adjustments were made depending on levels of parathyroid hormone (PTH), serum calcium, phosphorus, and calcitriol. No significant difference was seen between the groups in regard to age, sex, race, body mass index, dialysis duration, or cause of ESRD. Neither was any difference in the incidence of peritonitis seen between the groups. In the first 3-6 months, PTH decreased equivalently in both groups. The PTH level remained suppressed in the i.p. group throughout the remainder of the study, but, in the p.o. group, PTH returned to its pretreatment level after 3-6 months. Mean serum calcium was not different in the two groups. In the p.o. group, a considerably higher mean follow up phosphorus level (6.8 +/- 2.3 mg/dL versus 4.7 +/- 1.4 mg/dL, p = 0.008), PTH level (384 +/- 146 pg/mL versus 162 +/- 64 pg/mL; p = 0.005), and alkaline phosphatase level (178 +/- 37 IU/L versus 72 +/- 21 IU/L, p = 0.02) were seen as compared to the i.p. group. In the i.p. group, resolution of osteodystrophy occurred in all patients at the end of the study; in the p.o. group, 5 patients maintained or developed osteodystrophy by the end of the study (p = 0.016). We conclude that i.p. calcitriol is more effective than pulse p.o. calcitriol in lowering PTH and alkaline phosphatase levels and in resolving renal osteodystrophy, and that i.p. calcitriol is associated with a lower incidence of hyperphosphatemia and elevated Ca x PO4 byproduct. PMID- 11045317 TI - Vitamin B6 deficiency in elderly patients on chronic peritoneal dialysis. AB - Polyneuropathy is one of the most frequent manifestations in chronic uremia. Among the factors related to polyneuropathy, vitamin B6 deficiency is well known. The exact prevalence of vitamin B6 deficiency related to neurological manifestations has not been previously reported. We studied vitamin B6 status, collected self-reported symptoms, and carried out full neurological examinations in 66 patients on chronic peritoneal dialysis. Vitamin B6 status was estimated by direct measurement of pyridoxal phosphate. In general, symptoms related to vitamin B6 deficiency are peripheral neuropathies, such as paresthesia, burning and painful dysesthesias, and thermal sensations. These symptoms were reported and assigned one of five grade scores. Of our 66 patients, 12 patients complained at least one sensory abnormality. The levels of vitamin B6 in the patients varied between 1.0 ng/mL and 30 ng/mL. Patients who complained of neurological symptoms owing to vitamin B6 deficiency were significantly older than the other patients. In analyzing the symptomatic cases before and after vitamin B6 supplementation, a significant correlation was seen between the level of vitamin B6 and symptoms. Within one month after initiation of oral vitamin B6 supplementations (30 mg daily), levels of pyridoxal phosphate rose, and sensory abnormalities improved in 8 of 12 patients. When peripheral neuropathy is suspected in elderly patients on chronic peritoneal dialysis, vitamin B6 deficiency should be taken into consideration as the cause. If vitamin B6 deficiency is appropriately treated by oral supplementation, sensory abnormalities can be eliminated. PMID- 11045318 TI - Changes in lipid profiles in non diabetic, non nephrotic patients commencing continuous ambulatory peritoneal dialysis. AB - This study examined the effect on patient lipid profile of commencing continuous ambulatory peritoneal dialysis (CAPD). We followed eighteen non diabetic, non nephrotic patients for 9 months before and after dialysis commencement and compared lipid profiles. Mean cholesterol levels rose from 4.98 mmol/L to 5.42 mmol/L (p < 0.05). This change was chiefly due to a rise in low density lipoprotein (LDL) cholesterol. The LDL cholesterol rose after dialysis commencement and continued to rise up to 9 months later. High-density lipoprotein (HDL) cholesterol remained stable. Serum albumin and body weight fell during follow-up, suggesting that the rise in cholesterol was not a reflection of enhanced nutritional status. This study highlights the pro-atherogenic change in lipids that results from commencing CAPD. This phenomenon is not seen in hemodialysis, and it should be considered when selecting a dialysis modality, given the increased risk of cardiovascular disease in the dialysis population. PMID- 11045319 TI - Chest wall peritoneal dialysis catheter placement in infants with a colostomy. AB - The presence of a colostomy in infants with end-stage renal disease (ESRD) receiving peritoneal dialysis (PD) is associated with an inherent risk for contamination and the development of a PD catheter-associated infection. A two piece presternal catheter designed to reduce the incidence of such infections has been used in a small number of children, but the implantation of the catheter is technically difficult, and there is a risk of disconnection of the two parts secondary to rapid patient growth in the first year of life. Alternatively, a conventional Swan neck catheter, larger than typically required, can be placed with its exit site located on the chest wall. Over the past three years, we adopted this novel approach in two patients with ESRD and a colostomy in whom PD catheters were placed at ages 4 days and 12 days, respectively. During a combined follow-up of 50 months, only one episode of peritonitis and no episodes of exit site or tunnel infection have been observed. This experience supports the use of this unique approach to PD catheter placement in infants with ESRD and a colostomy. PMID- 11045320 TI - Impact of fill volume changes on peritoneal dialysis tolerance and effectiveness in children. AB - In the last decade, it has became apparent that the prescribed fill volume (IPV) in children on peritoneal dialysis (PD) should be expressed per body surface area (BSA in square meters) to avoid a false perception of peritoneal hyperpermeability as determined during a peritoneal equilibration test [PET, dialysate-to-plasma (D/P) ratio]. Nevertheless, the optimal IPV in terms of both tolerance and effectiveness remains under discussion. An individual approach to IPV prescription might balance the measurement of the intraperitoneal pressure, the use of the mass transfer coefficient despite the D/P ratio, and a determination of the effective peritoneal area available for exchanges. Considering these parameters, we usually found the individual optimal fill volume to be less than 1400 mL/m2. PMID- 11045321 TI - Nasal carriage of Staphylococcus aureus in families of children on peritoneal dialysis. European Pediatric Peritoneal Dialysis Study Group (EPPS). AB - Nasal carriage of Staphylococcus aureus is a risk factor for catheter-related infections with S. aureus in patients on chronic peritoneal dialysis (CPD). In children, S. aureus may transmitted to the catheter either from the patients' nares, or from the nares of caregiving carriers. As part of a prospective trial on the efficacy of mupirocin prophylaxis in children on CPD and their caregivers, we evaluated the prevalence of S. aureus carriage in 92 families of pediatric CPD patients. Patients and their caregivers (usually both parents) were screened by three nasal cultures obtained within four weeks. In 62% of the families, neither the patients nor any caregiver carried S. aureus. In 23%, the patient or at least one caregiver (sometimes both) was identified as a carrier. In 15%, at least one caregiver, but not the patient, was colonized with S. aureus. During further follow-up by once-monthly nasal cultures, 5 of the 57 initially negative patients developed S. aureus colonization, and in two families, at least one caregiver turned positive. Including these "occasional" carriers, the cumulative likelihood of one or several family members carrying S. aureus gradually increased to a plateau of about 55% after 6 observation months. Susceptibility rates of cultured S. aureus were 100% for vancomycin, 99% for aminoglycosides, 95% for piperacillin/tazobactam, 94% for cephalosporins, and 15% for ampicillin. In two patients and two caregivers (four different families), methicillin-resistant S. aureus was found. Three isolates from three different families were resistant to mupirocin. We conclude that S. aureus colonization is common in families of children on CPD. While 85% of carrier families are detected by 3 sequential nose cultures in patient and caregivers, up to 9 cultures may be required in "occasional" carriers. PMID- 11045322 TI - Dialysate cancer antigen 125 levels in children treated with peritoneal dialysis. AB - Peritoneal mesothelial cells are important for local host defense and membrane integrity. Dialysate cancer antigen 125 (dCA125) has been shown to be a good marker for the mesothelial cell mass in adult peritoneal dialysis (PD) patients. In children on PD, no information is available yet. We measured dCA125 in 65 dialysate samples from 24 PD children with a median age of 9.2 years (range: 2-18 years) and a median treatment time of 2.6 years (range: 0.1-9.3 years) on PD. The median dCA125 concentration was 8 U/mL (range: 2.3-30.7 U/mL), and the CA125 appearance rate (CA125AR) was 66.5 U/min/1.73 m2 (range: 18-282 U/min/1.73 m2). On cross-sectional analysis, a negative correlation was found between dCA125 and duration of PD treatment (r = -0.3, p = 0.04). No relation was found between age and dCA125 or CA125AR when the first measurement from each child was considered. No correlation was found between dCA125 and the mass transfer area coefficient of creatinine (MTACcreat). Longitudinal analysis showed a negative trend in CA125AR with duration of PD treatment (p = 0.03). No relation was found between peritonitis incidence and dCA125 or CA125AR. In conclusion, no influence of age on dCA125 and CA125AR was found. Levels of dCA125 declined with the duration of PD treatment, reflecting mesothelial cell mass, but they did not correlate with the MTACcreat or the peritonitis incidence in stable PD children. PMID- 11045323 TI - Severity of hyperemesis gravidarum correlates with serum levels of reverse T3. AB - To investigate the possible physiological relevance of extra-thyroidal production of reverse T3 (rT3) in hyperemesis gravidarum, measurements of serum rT3, free T3 (FT3), free T4, (FT4), and nonesterified fatty acids (NEFA) were correlated with weight loss of hyperemetic women. All the thyroid hormones, NEFAs and weight loss were significantly higher in hyperemesis gravidarum than in control subjects, and also higher than in those with milder symptoms of morning sickness (p < 0.05). Elevations of FT3, FT4 and NEFAs correlated with the extent of weight loss, the latter taken as the index of the severity of hyperemesis gravidarum (p < 0.05). Only rT3 correlated with both weight loss and the rate of lipolysis, as reflected by elevations of NEFAs (p < 0.05). The data are consistent with a shift from T3 to rT3 as products of 5'-monodeiodination of thyroxine in hyperemesis gravidarum. Because reverse T3 is physiologically inactive a control mechanism may be postulated wherein T3 production is minimized, thereby reducing weight loss and lipolysis in patients with hyperemesis gravidarum. PMID- 11045324 TI - The impact of early amniotomy on mode of delivery and pregnancy outcome. AB - OBJECTIVE: To evaluate the effect of early amniotomy in term gestation on the mode of delivery and pregnancy outcome in comparison with premature rupture of membranes (PROM) and oxytocin induction. STUDY DESIGN: The study population consisted of 60 consecutive parturients induced by early amniotomy. The two comparison groups were 147 women admitted with term PROM and 65 patients induced by oxytocin. All study participants were evaluated prospectively and had unfavorable cervical scores. RESULTS: The duration of the first stage of labor was significantly longer in the PROM group (987.8 +/- 572.3 min) as compared with the early amniotomy group (615.0 +/- 389.6 min) and the oxytocin induction group (650.9 +/- 349.5 min, P<0.001). Higher rates of CS were found in the study group (26.7%) as compared to the controls (11.6% in the PROM and 16.9% in the oxytocin groups, p=0.012). Neonatal outcome was similar in all groups. A stratified analysis comparing the risk of CS while controlling for a previous one did not show a significant difference between the early amniotomy and the oxytocin administration groups. CONCLUSIONS: Early amniotomy is associated with a higher rate of CS. While controlling for a previous CS, both ways of induction were comparable. In order to decrease the CS rates, induction should probably start with cervical ripening techniques in order to improve the Bishop scores. PMID- 11045325 TI - Cervical cancer in young Japanese women. AB - This study was performed to determine whether the incidence of cervical cancer in women aged 35 or younger has changed over the last 10 years and to examine the clinical characteristics of the cases. The incidence of cervical cancer in women aged 35 or younger were significantly greater in 1987-1991 than 1992-1996 (p = 0.001). Most new cases were detected by routine cytological screening. PMID- 11045326 TI - Serum leptin levels and the severity of preeclampsia. AB - The aim of this study was to evaluate the serum leptin levels in preeclampsia patients and in normotensive pregnant women, as well as, to assess an association with the severity of the disease. A cross-sectional study was carried out in 14 patients with mild preeclampsia, 12 with severe preeclampsia, and in 32 normotensive pregnant women during the third trimester of pregnancy. Rigorous criteria of selection were considered. The leptin levels were tested by an enzyme linked immunosorbent method. There were no significant differences in serum leptin concentrations between the patients with mild preeclampsia [13.6 +/- 11.2 (95% CI, 7.7-19.4) ng/mL], severe preeclampsia [14.8 +/- 11.5 (95% CI, 8.2-21.3) ng/mL] and normotensive pregnant women [12.5 +/- 7.9 (95% CI, 9.7-15.2) ng/mL]. In conclusion, serum leptin levels were similar in the patients with different grades of preeclampsia and normotensive pregnant women. PMID- 11045327 TI - Trends in epidemiology of preinvasive and invasive vulvar neoplasias. 13 year retrospective analysis in Thrace, Greece. AB - OBJECTIVE: To describe the trends in incidence of preinvasive and invasive vulvar neoplasias in the rural area of Trace, as well as to check the hypothesis that patients found with these two distinct entities represent populations with different epidemiologic characteristics. PATIENTS AND METHODS: We performed a retrospective study on 34 patients found with invasive vulvar cancer and vulvar intraepithelial neoplasia (VIN) (12 and 22 respectively), who referred to our department from 1986-1998. Epidemiologic characteristics of the patients were abstracted from medical charts. To evaluate our results we used the direct standardization method (1995 Eur. Population) and chi2 test. RESULTS: The age adjusted incidence rates for VIN1-2, in situ cancer (CIS)-VIN3 and invasive vulvar cancer were 0.9?100,000, 1.6?100,000 and 1.8?100,000 respectively, while for the whole VIN lesions 2.5?100,000. There were statistically significant differences in most epidemiologic characteristics between the two study groups, one with invasive cancer patients and one with patients found with preinvasive vulvar neoplasia. CONCLUSIONS: The overall age-adjusted incidence rate for vulvar neoplasia in Thrace is comparable to the one reported in literature. Our study results confirm that patients with invasive and preinvasive vulvar neoplasia represent populations with different epidemiologic characteristics. PMID- 11045328 TI - Acceptability of early transvaginal or abdominal sonography in the first half of pregnancy. AB - OBJECTIVE: To evaluate the acceptability of early transvaginal sonography by pregnant patients. METHODS: A questionnaire was completed by 246 patients and the resulting data were analysed. RESULTS: The incidence of discomfort was higher for the transabdominal than for the transvaginal route. Moreover, the transvaginal approach in early pregnancy, when compared with other transvaginal sonographic examinations was described as more satisfactory in 95% of cases. CONCLUSIONS: Transvaginal sonography in early pregnancy is better tolerated than transabdominal sonography. PMID- 11045329 TI - Obstetric and perinatal outcome of pregnancies with term labour and meconium stained amniotic fluid. AB - The purpose of this study was to evaluate the meconium staining of amniotic fluid (AF) in term of fetal distress, meconium aspiration syndrome, and perinatal morbidity and mortality. In a prospective study at Princess Badeea Teaching Hospital from April to November 1999, women with a singleton cephalic pregnancy of completed 37-42 weeks and with no pre-defined risk factor were recruited into the study. Study patients comprised 390 (10%) patients with meconium and 400 patients as controls but with clear amniotic fluid. Virtually meconium staining of the amniotic fluid was significantly associated with poor neonatal outcome in all outcomes measures assessed. Perinatal mortality increased from 2 per 1000 births with clear AF to 10 per 1000 with meconium (P<0.001). Other adverse outcomes also increased; e. g., severe fetal acidemia, Apgar score < or = 3 at 1 min and 5 min, and meconium aspiration syndrome. Delivery by cesarean section also increased with meconium from 7-14% (P<0.001). We concluded that meconium in the amniotic fluids associated with an obstetric hazard and significantly increase risks of adverse neonatal outcomes. Women with thin meconium in the presence of normal fetal heart rate can be safely managed at the clinical level. Mod-thick meconium alone should alert the obstetrician to a high risk fetal condition. Continuous fetal heart rate monitoring during labour and reassurance of fetal well-being by acid-base assessment were most significant factors in the reduction of meconium aspiration syndrome. PMID- 11045330 TI - Unicornuate uterus with a noncommunicating cavitary, laterally dislocated rudimentary horn presenting with adenomyosis, associated with ipsilateral renal agenesis. AB - Unicornuate uterus with a rudimentary horn is a rare mullerian anomaly. We report a case of a unicornuate uterus with adenomyosis in a laterally dislocated rudimentary horn, which showed a functioning endometrial islands, with associated ipsilateral agenesis of the kidney. PMID- 11045332 TI - Primary pelvic hydatid cyst. AB - We report a case of hydatid cyst of the pelvis in a 36-year-old woman presented with right adnexal cystic mass with similar cystic lesions in the liver. Laparatomy revealed a right paraovarian cystic mass densely adhered to the uterus, to the pelvic side wall, and to the right fallopian tube. Histopathological examination of the cyst wall showed the cuticular layer of the cyst. Cystic liver lesion was later proved to be hepatic hemangioma by magnetic resonance imaging. PMID- 11045331 TI - Delayed puberty associated with hyperprolactinemia caused by pituitary microadenoma. AB - Primary amenorrhea caused by the hyperprolactinemia is a rare condition characterized by the onset of thelarche and pubarche at appropriate ages but arrest of pubertal development before menarche. Hyperprolactinemia might be found in a few women with primary amenorrhea, yet relevant experience has apparently not been reported. We report a 16-year-old patient with hyperprolactinemia caused by a pituitary microadenoma. Her only symptom was delayed puberty without galactorrhea. Bromocriptine therapy was useful in order to induce the ovulation and cause the menarche. PMID- 11045333 TI - Leiomyomatosis peritonealis disseminata in a postmenopausal woman. AB - Leiomyomatosis peritonealis disseminata (LPD), a multifocal proliferation of smooth muscle like cells, has so far been described in 48 cases. Most of them were pregnant women or women taking the oral contraceptives, which supported the hypothesis that LPD is associated with hyperestrogenism. We report a 65-year-old women, without history of taking exogenous estrogens, which suggests that factors other than hormonal influences may contribute to pathogenesis of LPD. PMID- 11045334 TI - Case of hemorrhagic shock due to hypermenorrhea during anticoagulant therapy. AB - We report the case of a patient with uterine myoma who developed uncontrollable massive hemorrhage from the uterus during anticoagulant therapy after cardiac valve replacement and required hysterectomy. There was a discrepancy between the laboratory findings regarding the blood coagulation system and the clinical manifestations, suggesting a combination of multiple factors, such as a hormonal imbalance. This was a case that demanded strict attention to the management of the uterine lesions during the conduct of anticoagulant treatment. PMID- 11045335 TI - Hydrops of placental stem villi complicated with fetal congenital adrenal hyperplasia. AB - The authors present a case report of hydrops of placental stem villi. Numerous small aechoic spaces were demonstrated by prenatal ultrasonography. The patient spontaneously delivered a female newborn at 26 weeks' gestation. The infant showed hypertrophied clitoris and urogenital sius, and had a normal 46, XX karyotype. Endocrinological examination revealed that 3beta-hydroxysteroid dehydrogenase deficiency caused the anomaly. To our knowledge, this is the first report that congenital genital malformation complicated the placental mesenchymal dysplasia. PMID- 11045336 TI - Mullerian adenosarcoma of the uterus associated with tamoxifen therapy. AB - Mullerian adenosarcoma of the uterus is a biphasic tumor exhibiting benign epithelial and malignant stromal component. This tumor may occasionally be associated with tamoxifen therapy which is used as an adjuvant drug for breast carcinoma. Reviewing the literature, we found only 12 adenosarcoma cases associated with tamoxifen therapy Thus, the clinical and pathological findings in a 58 years old postmenopausal woman who developed uterine adenosarcoma, following low dose tamoxifen therapy after 8 years, was discussed in this report. PMID- 11045337 TI - Orthotopic liver transplantation for complicated HELLP syndrome. Case report and review of the literature. AB - A case of hepatic complication in a pregnant woman suffering from HELLP syndrome resulting in the need for transplantation is reported, and an algorithm for those instances is suggested. According to the literature, this is the 5th report of hepatic complications in HELLP syndrome necessitating liver transplantation. Since all but one of the transplanted patients survived (as opposed to a high mortality in non-transplanted patients), it is concluded, that a timely decision for transplantation is a safe option in this high risk group. PMID- 11045338 TI - Aachen keratoprosthesis as temporary implant for combined vitreoretinal surgery and keratoplasty: report on 10 clinical applications. AB - A new keratoprosthesis was used during pars plana vitrectomy in order to test the optical quality, watertightness, short-term biocompatibility and handling of the new device. The implantability was also tested, given that this keratoprosthesis might in future be left in place for several months. This Aachen keratoprosthesis (Aachen-KPro) is developed to be used as permanent implant to restore vision in corneal blind patients. PATIENT AND METHODS: The Aachen-KPro was used during pars plana vitrectomy in 10 patients with opaque corneas. In four cases, trauma precipitated the ocular disease. Eye burn was the cause of corneal and retinal disorders in another four cases. One patient had a history of congenital glaucoma with myopia, and one of uveitis with corneal dystrophy. After trephination of 6.5 mm in diameter, the Aachen-KPro, composed of soft silicone rubber, was temporarily placed in the trephination hole. After completion of the vitrectomy, the Aachen-KPro was replaced by a 7 mm corneal graft. RESULTS: Intraoperative use of the Aachen-KPro allowed uncomplicated intraoperative handling, smooth adaptation to the corneal rim in the trephination hole, and an undistorted view of the central and peripheral retina. Leakage, even during scleral depression, could be avoided by individual suturing of the scleral rim. After a follow-up period of 1-10 months, the retina was still attached in all cases. The corneal graft was clear after surgery in four eyes, and edema was found in three cases. Amnion or conjunctiva was placed over three patients' transplants. CONCLUSION: We report the first temporary implantations of a new keratoprosthesis in 10 patients. Its flexibility and good optical qualities allowed control of intraoperative procedures. The outcome and prognosis of the vitreoretinal surgery and keratoplasty were related to the primary diagnosis. The Aachen-KPro has shown advantages, especially in eyes where the anterior eye segment is severely damaged by eye burn or previous surgical interventions. In the future, prolonged use of the Aa-chen-KPro is planned for selected eyes. PMID- 11045339 TI - Multidrug resistance-associated proteins in glaucoma surgery. AB - BACKGROUND: Multidrug resistance (MDR) describes the phenomenon of cross resistance between different cytostatic agents which are structurally and functionally dissimilar. Two recently discovered proteins, lung resistance protein (LRP) and the multidrug resistance-related protein (MRP) have been implicated in the development of MDR. Since resistance to chemotherapeutic agents is a common problem in filtration surgery, especially in cases of complicated glaucoma, we decided to investigate the presence of MRP and LRP in surgically removed Tenon specimens from glaucoma patients. METHODS: The presence of MRP and LRP in surgically removed Tenon tissue (n=15) was analyzed by immunohistochemistry. The expression by cultured Tenon fibroblasts was assessed by reverse-transcriptase polymerase chain reaction (RT-PCR) and fluorocytometry. RESULTS: LRP expression was detected in 8 of 10 Tenon specimens. Positive staining for MRP was obtained in 5 of 10 specimens. Negative controls with non immune mouse IgG did not display any specific staining. RT-PCR and fluorocytometry revealed constitutive expression of MRP and LRP, at the RNA and protein level respectively, that was unaltered by pretreatment of the cells with mitomycin C or 5-fluorouracil. CONCLUSION: Our results demonstrate, that besides P-glycoprotein, other components of the MDR-system are present in conjunctival fibroblasts. Future developments in the use of chemotherapeutic agents in association with of filtration surgery need to take account of the presence of these counteracting mechanisms. PMID- 11045340 TI - The polyurethane nasolacrimal duct stent for lower tear duct obstruction: long term success rate and complications. AB - BACKGROUND: The polyurethane nasolacrimal duct stent is used as an alternative to conventional techniques for treatment of lower tear duct obstruction. The aim of this study was to evaluate the clinical success rate after a follow-up of 2 years. METHODS: Nasolacrimal duct stent implantation was attempted in 19 patients with nasolacrimal duct obstruction proven by digital substraction dacryocystography. The median age of the patients was 50 years, and the minimum duration of symptoms was 3 months. Patients were followed up 1 week, 6 months, 1 year and 2 years after the procedure. RESULTS: Eighteen stents were implanted in 17 patients without surgical complications. All stents were proven to be patent at the end of the procedure. Success rate, defined as proportion of patients free of symptoms, was 66.6%, 55.5% and 50% after 1 week, 6 months and I year, respectively, and remained unchanged thereafter. Three stents had to be removed between 6 months and 2 years after implantation. Histological examination showed granulation tissue growing into the opening and obstructing the stent in one case. CONCLUSION: Implantation of a polyurethane nasolacrimal duct stent is an alternative to conventional techniques in lower tear duct obstruction. Its overall success rate is lower than that reported after conventional dacryocystorhinostomy, but the procedure is fast, safe and reversible. Refinement of the surface and stent design may improve results in the future. PMID- 11045341 TI - Tear proteins of normal young Hong Kong Chinese. AB - BACKGROUND: Analysis of tear proteins is of diagnostic value for abnormal ocular conditions such as dry eye syndrome. Many studies of tear proteins have been performed on Caucasian subjects. However, little is known about these proteins in Chinese eyes. METHODS: The total tear protein concentrations of 30 normal young Hong Kong Chinese were determined by the Bradford method and the modified Lowry method. Bovine serum albumin (BSA) and bovine immunoglobulin G (IgG) were both used as standards for each method. The tear protein patterns were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the concentrations of major tear proteins were quantified by scanning densitometry after SDS-PAGE. RESULTS: The mean +/- SD total tear protein concentrations determined by the Bradford method, using BSA and IgG as standards, were 6.05 +/- 1.58 mg/ml and 11.48 +/- 2.32 mg/ml respectively. The values determined by the modified Lowry method, using the same two standards, were 9.66 +/- 2.03 mg/ml and 7.53 +/- 1.80 mg/ml respectively. The mean +/- SD concentrations of major tear proteins were 2.73 +/- 0.82 mg/ml for lactoferrin, 0.021 +/- 0.028 mg/ml for human serum albumin, 2.89 +/- 0.88 mg/ml for tear-specific prealbumin and 2.46 +/ 0.44 mg/ml for lysozyme. CONCLUSION: The results of total tear protein concentrations indicated that values obtained from different methods and different standards were not comparable. The tear protein patterns of our subjects were qualitatively similar to those reported for Caucasian subjects. However, the concentrations of the major proteins of our subjects were not in accordance with those reported previously. The main reason may be the large variability of method used. PMID- 11045342 TI - Three-dimensional analysis of measurements of the Heidelberg Retina Tomograph. AB - BACKGROUND: With the aid of scanning laser tomography, feasible with the Heidelberg Retina Tomograph (HRT), refined structures can be measured three dimensionally. Pictures are built up from scanned layers which are represented as two-dimensional topographical or reflectivity pictures by the HRT software. The kind of information that is provided by the third dimension can be exploited much better by real spatial three-dimensional presentation. The autostereoscopic Dresden 3D display makes such a spatial presentation feasible as add-on to the HRT. METHODS: Seventeen patients (9 women, 8 men) were chosen on the basis of a long duration of observation (mean 42.41 months) and a large number of follow-up examinations (mean 5.29). These patients were examined with the HRT as well as with the Dresden 3D display. The results were compared regarding their correlation pairs. RESULTS: Comparing the correlation pairs (r>0.7) the Dresden 3D display provided a higher Pearson correlation coefficient with 8 out of 10 pairs. It was evident that the evaluations of the two devices referred to the same parallel structures. DISCUSSION: The Dresden 3D display facilitates, with its form of presentation, a better evaluation of the measurements of the HRT. More precise evaluation of retinal structures by virtue of the autostereoscopic display presents an improvement. PMID- 11045343 TI - Polypoidal choroidal vasculopathy in Caucasians. AB - PURPOSE: To study the prevalence of polypoidal choroidal vasculopathy (PCV) in Caucasian patients with occult choroidal neovascularization (CNV); to study the clinical spectrum of PCV in Caucasians and the outcome after laser photocoagulation of such lesions. METHODS: (1) A consecutive series of 374 eyes of Caucasian patients at least 58 years old, presenting occult CNV, presumed to have age-related macular degeneration (AMD) on fluorescein angiography (FA) were further characterized by indocyanine green angiography (ICGA) to determine the frequency of PCV. (2) The funduscopic, FA and ICGA findings in a cohort of 36 Caucasian patients with PCV were analyzed. (3) The outcome after laser photocoagulation was studied in 14 PCV eyes with a minimum follow-up of 6 months. RESULTS: (1) Fourteen of 374 eyes (4%) presenting occult CNV in patients at least 58 years old were diagnosed as PCV by means of ICG-A. (2) A polypoidal lesion was found in the macula in 22 of 45 PCV eyes, in the peripapillary area in 16 of 45, under the temporal vascular arcade in 6 of 45 and in the midperiphery in 6 of 45. Large or soft drusen were observed in 15 of 45 eyes with PCV. (3) Regression of fundus signs without persisting polyps 6 months after laser photocoagulation was obtained in 5 of 5 treated peripapillary lesions but in only 5 of 9 treated macular or arcade lesions. CONCLUSION: Polypoidal choroidal vasculopathy is not rare in Caucasian patients presenting with occult choroidal neovascularization. The fundus abnormalities seen in such eyes overlap with the typical manifestations of AMD. Whereas the prognosis after photocoagulation of peripapillary polypoidal lesions appears to be relatively good, it is more guarded for macular or arcade lesions. PMID- 11045344 TI - Hereditary retinal dystrophies and choroidal neovascularization. AB - BACKGROUND: Choroidal neovascularization infrequently occurs in patients affected by hereditary retinal dystrophies. METHODS: We studied eight patients suffering from different hereditary retinal dystrophies (Best's disease, reticular dystrophy, butterfly-shaped dystrophy, gyrate atrophy, and retinitis pigmentosa) who developed choroidal neovascularization. All patients underwent complete ophthalmic evaluation, electrophysiology, colour vision testing, and fluorescein angiography. In some patients, ICG video-angiography was also performed. Laser treatment was carried out in only one patient. RESULTS: The mean duration of follow-up was 41.7 months (range 6-148 months). At CNV diagnosis, the mean VA was 0.23 (range 0.02-0.6). At the last follow-up, mean VA was 0.34 (range HM to 0.9). At the last follow-up, fluorescein angiography showed a focal, atrophic scar in seven eyes, a fibrotic membrane in two eyes and a still active membrane in two cases. CONCLUSION: We emphasize the relatively favourable visual prognosis in patients suffering from inherited retinal dystrophies complicated with choroidal neovascularization. Therapeutic approaches other than laser treatment could be attempted in these patients. PMID- 11045345 TI - Comparison of immersion ultrasound biometry and partial coherence interferometry for intraocular lens calculation according to Haigis. AB - BACKGROUND: The precision of intraocular lens (IOL) calculation is essentially determined by the accuracy of the measurement of axial length. In addition to classical ultrasound biometry, partial coherence interferometry serves as a new optical method for axial length determination. A functional prototype from Carl Zeiss Jena implementing this principle was compared with immersion ultrasound biometry in our laboratory. PATIENTS AND METHODS: In 108 patients attending the biometry laboratory for planning of cataract surgery, axial lengths were additionally measured optically. Whereas surgical decisions were based on ultrasound data, we used postoperative refraction measurements to calculate retrospectively what results would have been obtained if optical axial length data had been used for IOL calculation. For the translation of optical to geometrical lengths, five different conversion formulas were used, among them the relation which is built into the Zeiss IOL-Master. IOL calculation was carried out according to Haigis with and without optimization of constants. RESULTS: On the basis of ultrasound immersion data from our Grieshaber Biometric System (GBS), postoperative refraction after implantation of a Rayner IOL type 755 U was predicted correctly within +/- 1 D in 85.7% and within +/- 2 D in 99% of all cases. An analogous result was achieved with optical axial length data after suitable transformation of optical path lengths into geometrical distances. CONCLUSIONS: Partial coherence interferometry is a noncontact, user- and patient friendly method for axial length determination and IOL planning with an accuracy comparable to that of high-precision immersion ultrasound. PMID- 11045346 TI - Effects of scleral buckling of refraction and ocular growth in young rabbits. AB - BACKGROUND: There have been a series of reports indicating that scleral buckling (SB) surgery may induce high myopia in advanced retinopathy of prematurity. The mechanism of SB on refractive change in children, however, is not clearly known. We designed this study to investigate the effects of SB on refractive error and ocular growth in young rabbits and demonstrate their mechanisms. METHODS: For the study, SB surgery was performed on the right eyes of nine 5-week-old rabbits and 11 8-week old rabbits, with encircling buckle. The left eyes were monitored for control. Spherical equivalent, corneal power, and axial length were measured before SB and postoperatively at 2, 4, and 8 weeks. We compared the experimental group with the control group and analyzed the influence of age at the time of operation. RESULTS: In the control group of eyes, corneal power decreased, axial length increased, and spherical equivalent developed emmetropization with aging. In all eyes that underwent SB surgery, high myopia developed, with the increase in axial length as a major factor in inducing myopia. These changes were greater in the 5-week-old group than in the 8-week-old group. In the eyes that underwent SB surgery, axial length increased more than the eyes in the control group up to 2 weeks after surgery, but ocular growth was arrested after that time. CONCLUSIONS: SB in young rabbits effects ocular growth and results in high myopia, and an increase of axial length plays a major role in this mechanism. Also, the difference in the effects of SB according to age at the time of surgery may suggest that SB in premature infants could induce a significant axial myopia and secondary amblyopia. PMID- 11045347 TI - Retinal pigment epithelial autotransplantation: morphological changes in retina and choroid. AB - BACKGROUND: Replacement of degenerating retinal pigment epithelial (RPE) cells has been proposed to promote photoreceptor survival in retinal pigment epithelial disease. However, allografts of RPE have a higher rate of immunological rejection. The investigators studied the effects of autotransplantation of RPE on the morphological changes in photoreceptors and choriocapillaris. METHODS: RPE autotransplantation (wound type 1) was performed in 25 rabbits and compared with wounds following debridement of the RPE (wound type 2). Light and electron microscopy and scanning electron microscopy of choroidal vascular casts were performed at 3, 7, 14 and 30 days after surgery. Quantitative analysis of the choriocapillaris bed area was performed by automated image analysis and the results were analyzed by paired Student's t-test. RESULTS: Retinal pigment epithelium was found to be monolayered and ill-differentiated at day 7. Differentiation occurred at day 30, with preservation of photoreceptors and other layers of the neurosensory retina in type 1 wounds. Type 2 wounds showed atrophy of the regenerating retinal pigment epithelium at day 14 and day 30 with loss of overlying photoreceptor cell layer. There was significant choriocapillaris regeneration in type 1 wounds compared to the type 2 wounds at day 7 (P=0.045), day 14 (P=0.028) & day 30 (P=0.06). CONCLUSIONS: Retinal pigment epithelial autotransplantation significantly reduced photoreceptor degeneration and choriocapillaris atrophy associated with loss of retinal pigment epithelium. This procedure supports the hypothesis that RPE autotransplantation may be helpful in geographic atrophy and conditions requiring submacular surgery. PMID- 11045348 TI - Erbium-YAG laser-assisted preparation of deep sclerectomy. AB - BACKGROUND: Deep sclerectomy and viscocanalostomy are becoming more and more popular as non-penetrating filtering procedures. The purpose of the present study was to simplify the technique of this procedure and to reduce the rate of unintended perforations during the preparation of the deep lamella. METHODS: 20 enucleated porcine eyes were used. A superficial lamellar scleral flap with an area of 5x5 mm as for trabeculectomy was surgically prepared. Using a pulsed erbium:YAG laser the deep lamella (220 +/- 40 microm) with an area of 4x3 mm was removed. Ablation was performed with an energy of 40-100 mJ, a frequency of 1-10 Hz and a spot size of 500 microm and 1 mm (divergent beam). During the procedure the intraocular pressure was kept constant by continuous infusion. Finally the eyes were analyzed histologically. RESULTS: After initial trials it was possible to ablate the remaining deep corneoscleral lamella with the erbium:YAG laser without perforating into the anterior chamber. Starting with an energy of 70-85 mJ and a reduction to 40-60 mJ when reaching deeper layers, a spot size of 500 microm and a 10 Hz repetition rate gave the highest safety and efficiency in preparation. After a learning curve it was possible to preserve Descemet's membrane and intact trabecular meshwork in 10 consecutive operations as demonstrated by histology. CONCLUSION: Erbium:YAG laser-assisted deep sclerectomy offers an alternative to microsurgical preparation of the deep scleral lamella. The thermal damage is minimal (10-40 microm) and scarring may therefore not be stimulated. PMID- 11045349 TI - Ligneous conjunctivitis in a girl with severe type I plasminogen deficiency. AB - BACKGROUND: Ligneous conjunctivitis is a rare form of chronic recurrent pseudomembranous disease and may be associated with systemic membranous pathological changes. Recently ligneous conjunctivitis has been linked to severe type I plasminogen deficiency. We report on a patient with plasminogen deficiency and severe bilateral ligneous conjunctivitis. A new treatment approach and its outcome in this patient are described. CASE REPORT: We present the case of a 9 month-old Turkish girl with massive swelling of the eyelids and hard white pseudomembranes on both lids. The conjunctival smear was positive for Streptococcus pneumoniae. The clinical diagnosis was: ligneous conjunctivitis with superinfection. Histological investigation showed fibrin as major component of the pseudomembranes. The coagulation analyses revealed decreased plasminogen activity (<5%; normal 80-120%) and decreased plasminogen antigen (<0.4 mg/dl; normal 6-25 mg/dl). The failure of surgical therapy led to the attempt at treatment with intravenous lys-plasminogen. A significant improvement of the ocular symptoms occurred; stabilization with no recurrent pseudomembranes could be achieved for 6 months after treatment. DISCUSSION: The initial amelioration of symptoms in our patient after systemic replacement therapy confirms the etiological importance of plasminogen deficiency in the development of ligneous conjunctivitis. Curative treatment of ligneous conjunctivitis is still not available. However, intravenous application of plasminogen offers new possibilities in therapy, although long-term treatment seems necessary. PMID- 11045350 TI - Primitive neuroectodermal tumor of the orbit in a 5-year-old girl with microphthalmia. AB - PURPOSE: To report an orbital, intraconal, primitive neuroectodermal tumor (PNET) in a 5-year-old child with microphthalmia since birth. METHODS: Orbitotomy was performed and a large, polycystic, retroscleral, intraconal tumor was removed and subsequent histological, immunohistochemical and electron-microscopic analyses of the excised mass were performed. RESULTS: The tumor showed characteristic features of peripheral primitive neuroectodermal tumor including pseudorosettes, positive immunohistochemical reactions for the MIC2 gene and synaptophysin and ultrastructural finding of neurosecretory granules. CONCLUSION: This case is the first reported intraconal PNET of the orbit, and the first orbital case that expresses the MIC2 gene. In spite of the aggressive malignant features of peripheral PNET, the orbital variety seems to be the least aggressive since most of the reported patients are still alive. PMID- 11045351 TI - The ASCRS honouring of Albrecht von Graefe. American Society of Cataract and Refractive Surgery. PMID- 11045352 TI - Immunohistochemical localisation of two phosphatidylinositol 4-kinase isoforms, PI4K230 and PI4K92, in the central nervous system of rats. AB - The distribution and cellular localisation of the phosphatidylinositol 4-kinase isoforms, PI4K230 and PI4K92, that are believed to play important roles in the intracellular signalling mechanisms were studied in the rat brain (cortex, cerebellum, hippocampus and spinal cord) using immunocytochemistry with light and electron microscopy. PI4K230 was detected with a specific antibody purified by affinity chromatography from the egg yolk of chicken immunised with a 33-kDa fragment of bovine PI4K230, comprising amino acids 873-1175 of the native protein. PI4K92 was immunostained with a commercially available antibody raised in rabbit against amino acid residues 410-537 of human PI4K92. At the light microscopic level, the immunostaining of PI4K230 and PI4K92 showed a very similar distribution throughout the neurons and appeared as dense punctate labelling in the cytoplasm of perikarya and stem dendrites of various neurons. In addition to neurons, a strongly stained cell population was observed in the molecular layer of the cerebellar cortex that resembled Bergmann glia cells. Electron microscopy of neurons in the ventral horn of the spinal cord showed dense granular immunoprecipitates for both PI4K230 and PI4K92, mostly associated with the outer membrane of mitochondria and membranes of the rough endoplasmic reticulum. In addition, immunostaining of PI4K92 was also frequently found on the outer surface of cisterns and vesicles of Golgi complexes, whereas PI4K230 immunoreactivity was colocalised with some multivesicular bodies. Neither nuclear localisation nor a regular attachment to the cell membrane of these enzymes were observed. Our findings indicate that PI4K230 and PI4K92 are not involved directly in the ligand stimulated turnover of phosphoinositides at the plasma membrane of neurons. However, they may provide regulatory phosphoinositides for intracellular vesicular traffic being associated with various organelles. PMID- 11045353 TI - Three-dimensional spatial characteristics of caloric nystagmus. AB - We investigated the three-dimensional spatial characteristics of caloric nystagmus during excitation and inhibition of the lateral semicircular canal in five normal human subjects. Each subject was repositioned in 45 degrees steps at 1-min intervals such that the right lateral semicircular canal plane was reoriented in pitch, from 135 degrees backwards from the upright position to 135 degrees forwards, while the right ear was continuously stimulated with air at 44 degrees C. In orientations in which caloric stimulus resulted in excitation of the right lateral semicircular canal, the eye velocity axis was orthogonal to the average orientation of the right lateral semicircular canal plane. However, in orientations in which caloric stimulus resulted in inhibition of the right lateral semicircular canal, the eye velocity axis was orthogonal to the average orientation of the left and not the right lateral semicircular canal plane. These findings suggest that velocity and direction of caloric nystagmus depend not only on the absolute magnitude of vestibular activity on the stimulated side but also on the differences in activity between the left and right vestibular nuclei, most probably mediated centrally via brainstem commissural pathways. PMID- 11045354 TI - Dynamic vision based on motion-contrast: changes with age in adults. AB - Data are presented for a computerized test of dynamic vision in a sample of 1006 healthy subjects aged between 20 and 85 years. The test employed a form-from motion stimulus: i.e., within a random-dot display, Landolt rings of the same average luminance as their surroundings become visible only when the dots within the ring are moved briefly, while those of the surround remain stationary. Thus, detection of gap location is based upon motion contrast (form-from-motion) rather than luminance contrast. With the size and exposure duration of the centrally presented ring held constant, motion contrast was manipulated by varying the percentage (between 20 and 100%) of moving dots within the ring. Subjects reported gap location (left, right, top, bottom). A gradual decline of dynamic vision with age was found for all motion-contrast levels. Beyond 70 years of age, chance-level performance occurred in almost half of the subjects. The data provide the basis for applications including diagnostic screening for glaucoma, visual disturbances in brain-damaged patients, as well as assessment of the dynamic vision of drivers of motor vehicles and athletes. PMID- 11045355 TI - Proprioceptive population coding of two-dimensional limb movements in humans: I. Muscle spindle feedback during spatially oriented movements. AB - The proprioceptive coding of multidirectional ankle joint movements was investigated, focusing in particular on the question as to how accurately the direction of a movement is encoded when all the proprioceptive information from all the muscles involved in the actual movement is taken into account. During ankle movements imposed on human subjects, the activity of 30 muscle spindle afferents originating in the extensor digitorum longus, tibialis anterior, extensor hallucis longus and peroneus lateralis muscles was recorded from the lateral peroneal nerve using the microneurographic technique. In the first part of the study, it was proposed to investigate whether muscle spindle afferents have a preferred direction, as previously found to occur in the case of cortical cells, and to analyze the neural coding of the movement trajectories using a "population vector model." This model is based on the idea that neuronal coding can be analyzed in terms of a series of vectors, each based on specific movement parameters. In the present case, each vector gives the mean contribution of a population of muscle spindle afferents within one directionally tuned muscle. A given population vector points in the "preferred sensory direction" of the muscle to which it corresponds, and its length is the mean frequency of all the afferents within that muscle. Our working hypothesis was that the sum of these weighted vectors points in the same direction as the ongoing movement. The results show that each muscle spindle afferent, and likewise each muscle, has a specific preferred sensory direction, as well as a preferred sensory sector within which it is capable of sending sensory information to the central nervous system. Interestingly, the results also demonstrate that the preferred directions are the same as the directions of vibration-induced illusions. In addition, the results show that the neuronal population vector model describes the multipopulation proprioceptive coding of spatially oriented 2D limb movements, even at the peripheral sensory level, based on the sum vectors calculated from all the muscles involved in the movement. In an accompanying paper, the coding of more complex 2D movements such as those involved in drawing rectilinear and curvilinear geometrical shapes was investigated. PMID- 11045356 TI - Proprioceptive population coding of two-dimensional limb movements in humans: II. Muscle-spindle feedback during "drawing-like" movements. AB - It was proposed to study the proprioceptive sensory coding of movement trajectories during the performance of two-dimensional "drawing-like" movements imposed on the tip of the foot. For this purpose, the activity of the muscle spindle afferents from the Extensor digitorum longus, Tibialis anterior, Extensor hallucis longus, and Peroneus lateralis muscles was recorded from the lateral peroneal nerve using the microneurographic technique. The drawing movements, describing geometrical shapes such as squares, triangles, ellipses, and circles, were imposed at a constant velocity in both the clockwise and counterclockwise directions. A total number of 44 muscle-spindle afferents were tested, 36 of which were identified as primary and eight as secondary afferents. Whatever the shape of the imposed foot movement, the primary endings from one muscle never discharged throughout the whole trajectory (on average, they discharged for only 49.2% of the length of the trajectory), whereas all the secondary endings discharged for most part of the drawing trajectories (average: 84.8%). The relationship between afferent discharge rate and direction could be described with a cosine-shaped tuning function. The peak of this function corresponded to the preferred sensory direction of the receptor-bearing muscles. The whole path of a given geometrical drawing movement was found to be coded in turn by each of the primary afferents originating from each of the muscles successively stretched. The contribution of each population of muscle afferents from each ankle muscle was represented by a "population vector", whose orientation was the preferred direction of the muscle under consideration and whose length was the mean instantaneous frequency of the afferent population. The "sum vector" corresponding to the sum of all these weighted "population vectors" was found to point in the instantaneous direction of the drawing trajectory, i.e., the tangent to the trajectory. These findings suggest that trajectory information is already encoded at the peripheral level on the basis of the integrated inputs provided by sets of receptors belonging to all the muscles acting on a given joint. PMID- 11045357 TI - The three-dimensional vestibulo-ocular reflex during prolonged microgravity. AB - Single-case, longitudinal studies of the three-dimensional vestibulo-ocular response (VOR) were conducted with two spaceflight subjects over a 180-day mission. For reference, a control study was performed in the laboratory with 13 healthy volunteers. Horizontal, vertical and torsional VOR was measured during active yaw, pitch and roll oscillations of the head, performed during visual fixation of real and imaginary targets. The control group was tested in the head upright position, and in the gravity-neutral, onside and supine positions. Binocular eye movements were recorded throughout using videooculography, yielding eye position in Fick co-ordinates. Eye velocity was calculated using quaternion algebra. Head angular velocities were measured by a head-mounted rate sensor. Eye/head velocity gain and phase were evaluated for the horizontal, vertical and torsional VOR. The inclination of Listing's plane was also calculated for each test session. Control group gain for horizontal and vertical VOR was distributed closely around unity during real-target fixation, and reduced by 30-50% during imaginary-target trials. Phase was near zero throughout. During head pitch in the onside position, vertical VOR gain did not change significantly. Analysis of up/down asymmetry indicated that vertical VOR gain for downward head movement was significantly higher than for upward head movement. Average torsional VOR gain with real-target fixation was significantly higher than with imaginary-target fixation. No difference in phase was found. In contrast to vertical VOR gain, torsional VOR gain was significantly lower in the gravity-neutral supine position. Spaceflight subjects showed no notable modification of horizontal or vertical VOR gain or phase during real-target fixation over the course of the mission. However, the up/down asymmetry of vertical VOR gain was inverted in microgravity. Torsional VOR gain was clearly reduced in microgravity, with some recovery in the later phase. After landing, there was a dip in gain during the first 24 h, with subsequent recovery to near baseline over the 13-day period tested. Listing's plane appeared to remain stable throughout the mission. The findings reflect various functions of the otolith responses. The reduced torsional VOR gain in microgravity is attributed to the absence of the gravity dependent, dynamic stimulation to the otoliths (primarily utricles). On the other hand, the reversal of vertical VOR up/down gain asymmetry in microgravity is attributed to the off-loading of the constant 1-g bias (primarily to the saccules) on Earth. The observed increase in torsional VOR gain from the 1st to the 6th month in microgravity demonstrates the existence of longer-term adaptive processes than have previously been considered. Likely factors are the adaptive reweighting of neck-proprioceptive afferents and/or enhancement of efference copy. PMID- 11045358 TI - Age-dependence of malonate-induced striatal toxicity. AB - Malonate is an inhibitor of cellular metabolism, which, following intrastriatal injection, induces a striatal pathology similar to that seen in Huntington's disease. In two parallel studies, we have investigated the suggested relationship between the neuronal vulnerability to metabolic toxicity and the decline in metabolic function with increasing age. The first experiment investigated malonate-induced neuronal loss in animals aged from 6 weeks up to 27 months, and the second assessed the activities of two mitochondrial enzymes, succinate dehydrogenase and cytochrome oxidase (CYTOX) in animals aged 6 weeks, 3, 8 and 18 months. In the first study, male Lister-Hooded rats received intrastriatal stereotaxic injections of malonate (0.5 or 1.0 M). Animals were killed 10 days after surgery, and the brains were stained with cresyl violet and processed for NADPH-diaphorase activity and glial fibrillary-acidic-protein (GFAP) immunohistochemistry. Animals aged 6 months and older exhibited over 60% striatal neuronal loss. However, the degree of neuronal loss did not show any age-related increase in rats between 6 and 27 months of age, indicating that the extent of malonate-induced toxicity does not increase with age in animals older than 6 months. Infusion of 0.5 M malonate produced smaller lesions, which also demonstrated a consistent extent of neuronal loss from 6 months onwards. Metabolic enzyme activities were decreased in the striatum with increasing age, although this effect was only significant for CYTOX activity. Thus, the pattern of malonate-induced neuronal loss in aged animals partially reflects the changes in metabolic activity during ageing. PMID- 11045359 TI - Sound localization in hemispherectomized subjects: the contribution of crossed and uncrossed cortical afferents. AB - The aim of the present study was to evaluate how hemispherectomized subjects localize sounds in free field using residual auditory structures under monaural testing conditions. The main objective of using a monaural condition with these subjects, who lack the terminal fields of auditory projections on one side, was to evaluate how the crossed and uncrossed pathways compare, with the aim of resolving this biologically critical function. In this model, crossed and uncrossed inputs refer to auditory stimulation presented to the unobstructed ear on the contralateral and the ipsilateral side of the intact hemisphere, respectively. Three hemispherectomized subjects (Hs) and ten control subjects (Cs) were tested for their accuracy to localize broad band noise bursts (BBNBs) of fixed intensity presented on the horizontal plane. BBNBs were delivered randomly through 16 loudspeakers mounted at 10 degrees intervals on a calibrated perimeter frame located inside an anechoic chamber. Subjects had to report the apparent stimulus location by pointing to its perceived position on the perimeter. Hs were less accurate than Cs in the baseline binaural condition, confirming the finding that with a single hemisphere and/or residual (subcortical) structures they cannot analyze binaural cues to sound localization as efficiently as with two fully functional hemispheres. In the monaural condition, Hs localized poorly when they had to depend on the uncrossed input, but performed as well or even better than the Cs with the crossed input. These findings suggest that monaural spectral cues, which constitute the only residual cue to localization under the monaural testing condition, are treated more efficiently, that is, they lead to better localization performance when relayed to the cortex via crossed pathways than through uncrossed pathways. PMID- 11045360 TI - Disruptions in the reach-to-grasp actions of Parkinson's patients. AB - Previous research has shown that Parkinson's-disease (PD) patients produce irregular movement paths during a rapid arm pointing task. The aim of this study was to investigate the movement paths of PD patients during a prehensile action to objects requiring different levels of precision. Thus, we sought to determine if movement-accuracy requirements affect the control of movement path. Thirteen PD patients and 13 age-matched controls served as participants. In addition to having prolonged movement times, PD patients showed differences in the kinematic patterns of the transport and grasp components. For the transport component, relative time to maximum deceleration and relative time to maximum elbow velocity occurred earlier for the PD patients than the controls. Analyses of wrist paths indicated that, when accuracy requirements were increased, patients produced paths that appeared more segmented than controls. For PD patients, reaches to a small object resulted in wrist paths that were significantly less smooth, as reflected by higher jerk values, and were less continuous, as indicated by larger standard deviations in curvature. A temporal analysis of movement-initiation patterns in the vertical and horizontal planes indicated that control participants had a minimal offset between initial movement in the vertical plane and initial movement in the horizontal plane regardless of accuracy constraints. However, PD patients had a significantly longer interval between initial movement in the vertical plane and subsequent movement in the horizontal plane when reaching to the small object. Higher accuracy constraints also resulted in PD patients achieving relative time to maximum elbow velocity significantly earlier than controls. For the grasp component, PD patients produced movement patterns in which the amplitude of and relative time to maximum aperture were less sensitive to object size. In addition, patients exhibited greater variability in the time to maximum aperture. Additional analyses of the grasp component indicated that control participants exhibited a stable position, relative to object location, in which aperture began to close. Conversely, PD patients showed little consistency in where aperture began to close with respect to object location. Irregularities in the transport component suggest that PD patients have a reduced capability to precisely coordinate joint segments, particularly under high accuracy requirements. Variability in where aperture began to close and disruptions in transport-grasp coordination suggests that the basal-ganglia dysfunction, as exhibited in PD, affects the specification of these movement parameters used to produce a consistent pattern of coordination between prehensile components. PMID- 11045361 TI - Proprioception does not quickly drift during visual occlusion. AB - Several perceptual studies have shown that the ability to estimate the location of the arm degrades quickly during visual occlusion. To account for this effect, it has been suggested that proprioception drifts when not continuously calibrated by vision. In the present study, we re-evaluated this hypothesis by isolating the proprioceptive component of position sense (i.e., the subjects were forced to rely exclusively on proprioception to locate their hand, which was not the case in earlier studies). Three experiments were conducted. In experiment 1, subjects were required to estimate the location of their unseen right hand, at rest, using a visual spot controlled by the left hand through a joystick. Results showed that the mean accuracy was identical whether the localization task was performed immediately after the positioning of the hand or after a 10-s delay. In experiments 2 and 3, subjects were required to point, without vision of their limb, to visual targets. These two experiments relied on the demonstration that biases in the perception of the initial hand location induced systematic variations of the movement characteristics (initial direction, final accuracy, end-point variability). For these motor tasks, the subjects did not pay attention to the initial hand location, which removed the possible occurrence of confounding cognitive strategies. Results indicated that movement characteristics were, on average, not affected when a 15-s or 20-s delay was introduced between the positioning of the arm at the starting point and the presentation of the target. When considered together, our results suggest that proprioception does not quickly drift in the absence of visual information. The potential origin of the discrepancy between our results and earlier studies is discussed. PMID- 11045362 TI - Motor asymmetries in the human newborn are state dependent, but independent of position in space. AB - Human newborns have a preference for turning and maintaining the head to one side of the body. Most studies confirm a right-sided preference in supine. Few have addressed the state dependency of this lateral bias, and even fewer have examined whether it is also expressed in the semi-upright position. We investigated whether it varies as a function of behavioural state and position in space. Kinematic recordings of head movements were made with the newborn secured on a platform in the supine or semi-upright position, which alleviated biomechanical and postural constraints imposed by gravity. Newborns differed as to whether they had a vertex, Caesarean or breech delivery. The majority of infants maintained a right-sided preference in both positions, but it was strongly mediated by state. Delivery type did not account for any lateral bias. These findings provide convincing evidence that a lateral bias in movement and positioning of the head are reflections of active neural processes rooted in the regulation of state. PMID- 11045363 TI - Constraints on grip selection in cerebral palsy. Minimising discomfort. AB - The present study examined whether movement characteristics of individuals with spastic hemiparesis could be accounted for by disorders in movement planning. Two experiments were performed that tested minimisation of postural discomfort and minimisation of movement costs as constraints on grip selection. In the 1st experiment, spastic subjects and controls had to pick up a bar and place it in one of 5 boxes with either the left or right side down. In addition, awkwardness ratings of the different postures were given. Minimisation of posture discomfort as constraint on grip selection was examined. In line with previous studies, grip selection for the control subjects was based on minimisation of end posture discomfort. For the unimpaired hand of the spastic subjects, no discrimination was made in discomfort ratings among the different postures. Accordingly, with this hand, subjects showed no preference for a particular grip type. The posture ratings for the impaired hand were more varied both within and between spastic subjects, and minimisation of discomfort at either the start or end could not completely account for the grips chosen. Rather, the results suggest that minimisation of total posture discomfort acted as a constraint on grip selection for this hand. In the 2nd experiment, minimisation of total movement costs as a constraint on grip selection was tested. Spastic subjects and a control group had to grasp a bar on a clockface and rotate one end to the top position, starting from 15 different starting positions. Again, the end-state comfort effect was demonstrated in the control group. For the impaired hand of the spastic subjects, minimisation of total movement costs as expressed by a reduction of the total amount of joint rotation was shown to account for the grips chosen. The lack of consistency in grip selection for the unimpaired hand at some starting positions seemed to stem from an unresolved conflict between minimisation of end posture discomfort and minimisation of total movement costs. The combined results of the two experiments suggest that grip planning in spastic hemiparetic subjects is not disturbed per se. Rather, different constraints are imposed during the grip selection process. If movements are difficult to execute (i.e. because the hand is impaired), grip planning proceeded by a minimisation of total movement costs by choosing a task solution that reduced the total amount of joint rotation. PMID- 11045364 TI - Firing properties and dendrotoxin-sensitive sustained potassium current in vestibular nuclei neurons of the hatchling chick. AB - To understand the emergence of excitability in vestibular nuclei neurons, we performed patch-clamp recordings on brain slices to characterize the firing pattern on depolarization and the underlying currents in principal cells of the chick tangential nucleus. This study, on 0- to 3-day-old hatchlings, distinguishes electrophysiologically one main group of principal cells based on their response to depolarizing current pulses (300-400 ms) in current-clamp recordings. This group (90%; n=29) displayed nonaccommodating, repetitive firing on depolarization. The remaining cells fired one action potential at the beginning of the current pulse and then accommodated. In voltage-clamp recordings, a low-threshold, sustained, dendrotoxin-sensitive (DTX; 200 nM) potassium current, I(DS), was identified in both cell groups. In the repetitively firing principal cells, the mean proportion of the DTX-sensitive sustained current contributing to the total outward current was less than 20%. This percentage is significantly less than that reported (45%) in a previous study performed in late chick embryos (E16), in which most of the cells (83%; n=89) were accommodating neurons. Tonic firing is an important electrophysiological feature characterizing most mature, second-order vestibular neurons, since it allows the neurons to process signals from behaviorally relevant inputs. Accordingly, this study contributes toward defining the emergence of the mature pattern of neuronal excitability and the ionic currents involved. PMID- 11045365 TI - Saccular and utricular influences on sympathetic nerve activities in cats. AB - We studied the effects of stimulation of the utricular and saccular nerves on sympathetic nerve activity in decerebrated cats. Bipolar electrodes were fixed in place on the utricular and/or saccular nerve under visual observation; the other branches of the vestibular nerve were transected. Baroreceptors and vagus nerves were inactivated bilaterally so that inputs from baroreceptors and other visceral receptors did not influence the sympathetic nerve outflow. Postganglionic sympathetic nerve activity was recorded from the renal branch of the sympathetic nerve, which is known to be more sensitive to vestibular stimuli than other types of sympathetic fibers. With stimulation of either the saccular or utricular nerve at low stimulus intensity, a prominent inhibition followed by a rebound excitation was evoked on spontaneous renal nerve discharges. The latency of the inhibition ranged from 65 to 130 ms, and the duration of inhibitory responses was about 90-150 ms. An increase in stimulus intensity in both the saccular and utricular nerves induced inhibitory effects preceded by short-term excitation. The latency of this excitation, which was superimposed on the initial phase of the inhibitory responses, ranged from 55 to 90 ms. PMID- 11045366 TI - Dental caries in adult Lithuanians. AB - There are few data on the incidence of dental caries in Lithuanian adults. The aim of the present study was to describe caries and treatment experience among 35 44 and 65-74-year-olds, and to relate this to certain selected independent variables (gender, urban/rural residence, drinking water fluoride levels, and years of education). A total of 680 subjects selected based on a stratified random sampling procedure (response rate 52%) were examined by one examiner. Dental caries was recorded as DMFT following the WHO recommendations. The results showed that the median DMFT scores were 18 for the 35-44-year-olds (n = 380) with median DT = 2, MT = 5, FT = 7. For the 65-74-year-olds (n = 300) the median DMFT was 24, with DT = 1, MT = 18, FT = 2, respectively. One percent of all 35 year olds and 11% of 65-74-year-olds were edentulous. In the younger age group, statistically significant differences in the DMFT scores were related to gender, urbanization and drinking water fluoride levels. Participants from areas with high fluoride content in the drinking water (> 1.5 ppm F/1) had lower DT, MT, and FT values. Females and participants from urban areas had higher numbers of FT. Participants with more years of education had lower DT, MT, and higher FT values. In the elderly, DMFT scores were related to water fluoride levels and years of education. Individuals with more years of education had higher numbers of FT and lower MT values in this age group. Poor oral hygiene was associated with high numbers of DT in both age groups. The data indicate that dental caries is widespread among adult Lithuanians. PMID- 11045367 TI - Assessment of the validity and consequences of different methods of expressing the severity of dental fluorosis in a subject. AB - The aim was to assess the validity and consequences of different methods of expressing severity of dental fluorosis in a subject. The analyses were based on Ugandan children (n = 481), aged 10-14 years, with life-long consumption of drinking water with either 0.5 or 2.5 mg fluoride per liter. Fluorosis was assessed using the Thylstrup and Fejerskov (TF) index. All children (n = 219) with 28 teeth and fluorosis on at least 1 tooth pair were selected to test methods of expressing fluorosis, e.g. the TF score on the most severely affected tooth, a maxillary central incisor, the median and the mean scores. A test group (n = 40), the 10 most and the 10 least severely affected children in each district, was used to evaluate the methods and a reference group (n = 179) to confirm or refute the findings in the test group. To evaluate consequences of the different methods of expressing severity of fluorosis in a subject, children from the low (n = 130) and the high fluoride (n = 132) districts not included in the test or reference group formed the community comparison groups. Comparison between the median (gold standard) and mean scores showed a significant deviation in the reference group only. Most of the partial recording methods, such as the score for the most severely affected tooth, were significantly and systematically higher than the median. While for all recording methods the median score was zero in both communities, the distribution of the subjects according to severity differed significantly between the communities. PMID- 11045368 TI - Effects of a high sucrose diet and intragastric sucrose feeding on the dentinogenesis, dental caries, and mineral excretion of the young rat. AB - Previous studies show that a high sucrose diet reduces the rate of primary dentinogenesis and increases dental caries, although their cause-effect relationship is still obscure. The purpose of this study was to explore whether the effect of sucrose load on the dentinogenesis and dental caries of young rat molars is mediated by systemic (intragastric) or by systemic and local (dietary) factors. At weaning (19 days), animals were randomized into the control, intragastric sucrose, and dietary sucrose groups for 4 weeks. The areas of dentin appositions and dentinal caries lesions were measured planimetrically. Caries was also determined with Shiffs staining and the width of predentin by histology. Urinary Ca, K, and Na levels were measured by flame photometry, urinary P levels using an UV method, and serum insulin levels using radioimmunoassay. Systemic and local sucrose load reduced dentin appositions and intragastric sucrose increased urinary Ca excretion. No differences in the width of predentin were noticed. Only dietary sucrose enhanced the occurrence and progression of caries. The present findings show that sucrose load reduces dentinogenesis by impairing the synthesis of dentin matrix, but also point out the crucial importance of the local sucrose challenge in the initiation of dental caries. PMID- 11045369 TI - Correlations between total protein, lysozyme, immunoglobulins, amylase, and albumin in stimulated whole saliva during daytime. AB - The correlations between salivary proteins and the daytime variations are not known. The present study investigated the within-subject variation of correlations and concentrations between lysozyme, IgA, IgG, IgM, albumin, amylase, and total protein in stimulated whole saliva of healthy adults in the course of a 12-h period. After several practise sessions, unstimulated and stimulated whole saliva samples were collected five times daily (at 8 a.m., 11 a.m., 2 p.m., 5 p.m., and 8 p.m.) from 30 healthy university students. Flow rate and total protein concentration were used as covariates, and gender as a between subject factor in the MANOVA analysis. After this adjustment, there was significant within-subject variation in salivary IgA (P < 0.001), albumin (P < 0.01), amylase (P < 0.05), and total protein (P < 0.001) concentrations. Total protein correlated significantly with amylase albumin and IgA through different samplings. In addition, IgG correlated with albumin and lysozyme in the course of 12 h. On the whole, the correlations between variables remained stable during repeated samplings. In addition, rankings of subjects for the variables tended to be maintained across different samplings (P < 0.001). However, the observed within-subject variations in salivary IgA, albumin, amylase, and total protein concentrations suggest that these proteins are subject to short-term variation. PMID- 11045370 TI - Comparison of oral hygiene efficacy of one manual and two electric toothbrushes. AB - The purpose of the study was to compare the efficacy of two electric toothbrushes (Philips HP555 and the Philips Jordan 2-action Plaque Remover HP510). A manual toothbrush Jordan V-shaped, medium) served as control. Fifty subjects, aged 18-60 years, participated in a randomized, single-blind, 3 x 3 weeks crossover study. Plaque was assessed according to the Turesky modification of the Quigley-Hein index (P.I.), while the Loe-Silness index was used for assessing gingivitis. Adverse effects were assessed according to the ADA specifications. Compliance and preference were assessed through questionnaires and interviews, respectively. All periods mean P.I. (all surfaces) were 2.79, 3.01, and 2.86 for the manual, the HP555, and the HP510 electric brushes and the corresponding gingivitis values were 1.19, 1.22, and 1.21. For both indices, only the difference between the manual and the HP555 yielded significance (P = 0.04 and P = 0.02). Most subjects (28/50) preferred the HP5 10 brush, as it felt more practical to use and was perceived to have better cleaning ability. In conclusion. no clinically relevant differences in plaque reducing and gingivitis controlling ability were observed. PMID- 11045371 TI - Type X collagen in human enamel development: a possible role in mineralization. AB - Although type X collagen is one of the key molecules in endochondral ossification, no data are available on whether it is present in dental structures when mineralization is proceeding. We therefore monitored the appearance of type X collagen in tooth germs of human samples ranging in gestational age from 17 week-old fetuses to 9-week-old newborn. Using immunohistochemistry, ELISA techniques, and Western blotting, we show that type X collagen is present in human tooth germ during enamel maturation. Intense immunohistochemical staining for collagen type X was observed in the enamel and in the apical parts of secretory ameloblast at the bell stage when the dentine and enamel matrix were already under formation. The odontoblasts, the dentine, and the pulp were not stained. In the early (9-week) postnatal stage, the staining for collagen type X in the enamel matrix was diminished, and only a very weak signal could be detected in the secretory ameloblasts. A positive reaction for collagen type X was also observed in ELISA assay of extracts obtained from human embryonic enamel and hypertrophic cartilage samples. The Western blot analysis of the enamel demonstrated that size of the molecule detected by MoAb X53 is characteristic of the type X collagen. This correlates well with our immunohistochemical findings. Based on these data, we propose that type X collagen is one of the candidate molecules present in the enamel matrix that might be involved in mineralization of the enamel. PMID- 11045372 TI - Trends and prognoses of dental status in the Swedish population: analysis based on interviews in 1975 to 1997 by Statistics Sweden. AB - The aims of this study were to describe changes in dental status over the 22-year period from 1975 to 1997, and to make a prognosis of dental status based on these data for the years 2005 and 2015. The study is based on regular investigations of the living conditions performed by Statistics Sweden of samples varying between 11,582 and 14,964 participants and a response rate from 78% to 86%. The questions of the interview used in this study were focused on dental status and utilization of dental services. The prevalence of edentulism in the age group 25-74 years decreased from 19% in 1975 to 3% in 1996/97. The proportion of dentate persons increased from 75% in 1975 to 97% in 1996/97 in age group 45-64 years with similar trends in the other age groups. In 1996/97, 2.1% of the whole sample (16 84 years) reported that they had received implant-supported restorations. The rate was higher among the elderly and the edentulous subjects. The great regional differences in dental status found in the first part of the observation period remained only in the oldest age group in 1996/97. The prognosis predicts that 95% of the subjects in age group 65-74 years and 90% in age group 75-84 years will be dentate in the year 2015. The substantial increase of dentate subjects among the elderly that has occurred during the past few decades and its expected continuation in the coming years implies a great change in need and demand for dental care services. PMID- 11045373 TI - Genetic diversity of Porphyromonas gingivalis and its possible importance to pathogenicity. AB - During recent years much effort has been put into understanding the genetic composition of the oral populations of black-pigmented anaerobic bacteria. One of them, Porphyromonas gingivalis, is a putative periodontopathogenic organism considered to be particularly relevant in the etiology of adult periodontitis. It has been shown in studies using molecular typing methods that most bacterial populations consist of numerous genetic clones, and that only a small proportion of these clones cause disease. Elucidation of a possible association of genotypic profiles with either disease or clinical healthy condition is important for understanding the pathogenic characteristics of bacteria. Studies addressing this issue as it relates to P. gingivalis are reviewed in the present article. Genotypic characterization of P. gingivalis strains has revealed extensive heterogeneity in natural populations of this bacterium. Some of the potential virulence factors of P. gingivalis have been purified and cloned and methods have been established to identify their genes. Although no studies have clearly defined the relationship between a specific genotype of P. gingivalis and periodontal status of the host, it seems that molecular typing tools, which are undergoing rapid improvements, will allow us to distinguish between virulent and avirulent strains of the same species in the near future. PMID- 11045374 TI - Molecular biology of human immunodeficiency virus type 1. AB - Since the discovery of human immunodeficiency virus (HIV) as the etiologic agent of acquired immune deficiency syndrome (AIDS) more than a decade ago, tremendous progress has been made in various aspects of this virus and its interplay with the host immune system. The advent of potent combination therapy has made it possible to achieve effective and durable control of HIV-1 replication in vivo, yet the persistence of the latent reservoirs pose a new challenge. The recent identifications of several cellular proteins interacting with different viral gene products have not only shed new insights into our understanding of the HIV-1 and the host cell biology, but also provided the bases for developing novel strategies to block HIV-1 replication. It is from this perspective that we review the current understanding of the molecular mechanisms governing the HIV-1 life cycle. PMID- 11045375 TI - Different forms of HSV-1 VP22a within purified virion and infected cells. AB - Utilizing the monoclonal antibody MCA406, our experimental data suggest that the herpes simplex virus type 1 (HSV-1) protein, VP22a, is present in the purified virus in a different form from that present within infected cells, namely the virion and infected-cell form, respectively. It seems reasonable to suggest that two different forms of VP22a are synthesized during the HSV-1 productive cycle. Using varying quantities of reducing agents, both inter- and intramolecular disulfide linkages were demonstrated in this protein family. Moreover, the VP22a virion form could not be detected under nonreducing conditions by monoclonal antibody, even in the presence of proteolysis inhibitors, i.e. aprotinin, phenyl methane-sulfonyl fluoride (PMSF) and soybean trypsin inhibitor. Varying temperature had little effect on the breakdown of VP22a disulfide bonds. A higher molecular-weight band, present in the nonreduced gel tracks, clearly indicates the presence of intermolecular disulfide bonds. Similarly, the appearance of bands of lower apparent molecular weight in the nonreduced tracks suggests the presence of intramolecular disulfide bonding. The VP22a infected-cell form may be modified to the virion form during the capsid-assembly process, prior to full capsid formation. PMID- 11045376 TI - Genotypic analysis of HCV infection in kinmen. AB - In this report, stratified random sampling of an epidemiological study population from the town of Kin-Hu in the Kinmen Islands was used to create a subpopulation of 832 individuals. Two enzyme immunoassays (EIA) were used for antibody testing including Abbott's hepatitis C virus (HCV) EIA 2nd Generation and a synthetic peptide-EIA, NANBASE C-96-EIA, based on the locally predominant strain of HCV. In addition to RIBA and DBL immunoblot assays, reverse transcription-polymerase chain reaction (RT-PCR) was employed to confirm HCV infection. Results showed that 20 of 832 (2.4%) adults in Kinmen had HCV infection. In terms of genotype distribution, 31.3% (5/16) were infected with both 1a and 1b genotypes, 25.0% (4/16) only with the 1b genotype, and 43.8% (7/16) with the 2a genotype. Through comparative analysis of RT-PCR, RIBA, and DBL results, we found that the sensitivities of RIBA and DBL could safely be increased by modifying the definition of a positive case. If the presence of a reactive band with > or = 2+ antibody reactivity to core protein is accepted as positive for overall RIBA or DBL testing, sensitivity is increased without adversely effecting specificity. PMID- 11045377 TI - Effects of quorum sensing signal molecules on the hydrogen peroxide resistance against planktonic Pseudomonas aeruginosa. AB - The effects of quorum sensing signal molecules in Pseudomonas aeruginosa, N butanoyl-L-homoserinelactone (C4-HSL) and N-(3-oxododecanoyl)-L-homoserinelactone (3-oxo-C12-HSL) on planktonic cell resistance against hydrogen peroxide were studied. In P. aeruginosa JP2 cells with the deletion of lasI and rhlI, the viable cell concentration decreased with time and was reduced by about 4 log after 2 h of 7.5 mM H2O2 treatment, while only a 2-log reduction was found for the wild type P. aeruginosa PAO1 cells. When cultured with 20% PAO1 spent medium, P. aeruginosa JP2 showed similar hydrogen peroxide resistance to that seen in P aeruginosa PAO1. Culturing with 20% JP2 spent medium or with 10 microM C4-HSL and 20 microM 3-oxo-C12-HSL did not affect P aeruginosa JP2 cell susceptibility to hydrogen peroxide. Although both 20% PAO1 and JP2 spent media reacted with H2O2 and reduced H2O2 to 50% of the strength of the original concentration, the remaining H2O2 was still sufficient to kill P. aeruginosa JP2. These results indicate that the difference in cell resistance against H2O2 between P. aeruginosa PAO1 and JP2 was related to the existence of gene products of the lasI and rhlI systems. However, adding synthetic homoserine lactones alone did not increase P. aeruginosa JP2 cell resistance to H2O2 as seen in the experiments adding PAO1 spent medium. Determination of the detailed relation between cascade regulation in P. aeruginosa and its cell resistance to H2O2 will require further investigation. PMID- 11045378 TI - Costimulatory molecules expression and cytokine profiles of cord blood mononuclear cells in newborns with low and high risk of developing atopic diseases. AB - This study sought to determine predictors of atopic diseases in newborns. We evaluated the levels of expression of costimulatory molecules (CD80 and CD86) and the production of cytokines [interleukin (IL)-12, interferon (IFN)-gamma, IL-4, IL-10] in the cord blood of mononuclear cells in high risk newborns (n = 17), and compared them with those in low risk newborns (n = 25). Fluorescence-activated cell sorter (FACScan) analysis was performed to determine the expressions of CD80 and CD86 on activated B cells and monocytes of both groups. The levels of IL-10, IL-12p40 and IL-12p70 in the supernatant were assayed by enzyme-linked immunosorbent assay (ELISA), and also the mRNA levels by reverse transcription polymerase chain reaction (RT-PCR). Intracellular staining of IL-4 and IFN-gamma in stimulated mononuclear cells was also performed as well. The expressions of CD80 and CD86 on B cells showed no significant difference between the high and low risk group. There was greater expression of CD86 on the monocytes of low risk newborns as compared to high risk newborns (p < 0.05). When B cells and monocytes isolated from the cord blood of both groups were stimulated with mitogens, the production of IL-10, IL-12p40, and IL-12p70 in the supernatants was not significantly different. The expressions of mRNA of IL-10, IL-12p35, and IL 12p40, and the intracellular staining of IL-4 and IFN-gamma in stimulated mononuclear cells were not significantly different between the two groups. These findings suggested that cytokine profiles in the cord blood cannot predict the development of atopic diseases. Determination of whether preferential expression of costimulatory molecules is of predictive value or not will require further study. PMID- 11045379 TI - Evaluation of different blood culture media in neonatal sepsis. AB - Blood drawn from neonatal patients is available in only limited amounts for blood cultures. The BACTEC Pediatrics (PEDS) Plus aerobic nonradiometric blood culture bottle was designed to aid the diagnosis of pediatric bacteremia. To assess the value of PEDS Plus blood culture medium for neonatal patients, we prospectively compared the PEDS Plus blood culture medium with the standard BACTEC aerobic medium (NR6A). From January to December 1999, 192 pairs of PEDS Plus and NR6A blood culture bottles were collected by the pediatric microbiology laboratory of Taipei Veterans General Hospital. Seventeen (8.85%) isolates were considered to be clinically significant microorganisms. Isolation rate of the PEDS Plus culture bottles was significantly higher than that of the NR6A culture bottles (p < 0.001). Six isolates were detected at least 1 day earlier by using the PEDS Plus culture bottles (p < 0.001). Among those patients who were under antibiotic therapy, the PEDS Plus culture bottle showed more significant growth than the NR6A culture bottle (p < 0.001). After weighing their benefits, we suggest substituting PEDS Plus bottles for NR6A bottles when culturing blood from neonatal patients or from those who are receiving antibiotic therapy. PMID- 11045380 TI - Invasive Streptococcus pneumoniae infections of children in central Taiwan. AB - We carried out a retrospective study on childhood invasive pneumococcal infections (IPI) diagnosed from the January 1990 through the April 2000 at a medical center in central Taiwan. Their clinical features, outcome of the patients and the resistance patterns of the isolates were analyzed. A total of 95 clinical isolates from 72 patients younger than 14 years of age were included in this study. Of these 72 patients, 51 had bacteremia, 28 meningitis, 14 bacteremic pneumonia, 12 pleural empyema, eight otitis media, four arthritis, three sinusitis, two periorbital abscesses, one deep neck infection, one psoas muscle abscess, one peritonitis, one urinary tract infection, and one cutaneous infection. Ancillary diagnostic tests, including Gram stain smears and latex agglutination tests, were applied and the sensitivities were 86.2% and 54.3%, respectively. The prevalence rate of penicillin nonsusceptible Streptococcus pneumoniae has increased dramatically since 1995 in central Taiwan, with rates of 5.6% and 74.1% before and after 1995, and the overall mortality rate was 20.8% and 53.3% respectively. Ten of 19 children (52.6%) with pneumococcal meningitis who survived had long-term sequelae. PMID- 11045381 TI - Enteric adenovirus infection in children in Taipei. AB - Enteric adenoviruses (EAds), including type 40 (Ad40) and 41 (Ad41), can cause acute and severe diarrhea in young children. To delineate the epidemiological features of pediatric EAds infection in Taiwan, we conducted a retrospective study of all cases of EAds gastroenteritis in children treated at National Taiwan University Hospital for the period from July 1993 to December 1997. Stool samples were tested for the presence of Ad40 or Ad41 by enzyme immunoassay (EIA). A total of 64 cases of EAds infection in 63 children aged from 8 days to 81 months old with a median age of 9.5 months treated during the study period were identified. The male-to-female ratio was 1.63 (39/24). No obvious seasonal clustering of EAds cases was noted. Most patients (76.6%) were infected before the age of 2 years. Clinical features included diarrhea (96.9%), fever (54.7%), vomiting (45.3%), mild dehydration (43.8%), symptoms of upper respiratory tract infection (21.9%), and abdominal pain (12.5%). Analysis of fecal samples in patients with diarrhea showed watery diarrhea in 72.2%, diarrhea with mucus in 20%, diarrhea with blood in 3.1% and diarrhea with mucus and blood in 1.6 % of all patients. Nearly one half (43.5%) of the patients had diarrhea for more than 7 days. Thirty-seven patients (57.8%) were hospitalized due to gastroenteritis or other unrelated diseases, and 11 patients (17.2%) acquired enteric adenovirus infection during hospitalization for other underlying disease. Twelve patients (18.8%) had mixed infections, which included rotavirus, respiratory syncytial virus (RSV) and Salmonella species. There were no deaths in this series. The findings of this study suggest that EAds are important etiologic microbes of pediatric gastroenteritis. PMID- 11045382 TI - Rotavirus gastroenteritis in children: 5-year experience in a medical center. AB - Rotavirus infection is the leading cause of childhood gastroenteritis. We retrospectively reviewed cases of rotavirus gastroenteritis at National Taiwan University Hospital from January 1993 to December 1997. During the study period there were 429 patients with rotavirus infection with ages ranging from 1 day to 16 years with a median of 13 months. The male-to-female ratio was 1.2:1. Infection occurred before the age of 2 years old in 76% of patients. The seasonal peak occurred in the late winter and early spring during 1993 to 1996, but the case number increased in late spring and summer in 1997. The G serotype of the rotavirus was identified in 302 patients (70%). Vomiting and dehydration developed more frequently following infection with G1 rotaviruses, while an increased frequency of seizures was noted following G2 infection; the differences were not statistically significant. One patient had two episodes of infection; the first one was caused by G1 rotavirus, and the strain causing the second infection could not be typed. In conclusion, the results suggest that there is a strong seasonal variation in the incidence and characteristics of rotavirus infection in Taipei area. The infections caused by G1 and G2 rotaviruses were clinically indistinguishable. PMID- 11045383 TI - Clinical presentation of acute mastoiditis in children. AB - Nineteen children with 21 episodes of acute mastoiditis were treated in our hospital from 1989 to 1998. The diagnosis was based on physical, radiologic, and surgical findings. The affected children were aged from 1 year old to 17 years old, with the peak incidence at 4 years old (23.8%). Postauricular pain (90.5%) and fever (81%) were the most common harbingers of incipient acute mastoiditis. Streptococcus pneumoniae (38.1%) was the most common organism isolated followed by Pseudomonas aeruginosa (23.8%). Underlying diseases such as leukemia and myeloid metaplasia were found in 38.6% of patients. All of the patients were initially treated with intravenous antibiotics during hospitalization. Six patients were managed with an adjunctive drainage procedure such as myringotomy or mastoidectomy. The most common complication of acute mastoiditis was hearing loss (31.6%); the second was meningitis (21.1%). Subperiosteal abscess was found in two patients and brain abscess in one. Although acute mastoiditis is an uncommon condition, early diagnosis and management are necessary to prevent more serious complications. PMID- 11045384 TI - Tetanus of the elderly. AB - The medical records of 20 tetanus patients who were treated at a university teaching hospital in Taiwan during the period from October 1991 to July 1999 were retrospectively analyzed. There were 18 adults (six males and 12 females) with unknown previous immunization status and ages ranging from 34 to 87 years old (mean 63 years). Two patients were children, aged 3 and 5 years old, respectively; both of them had incomplete tetanus immunization. Of the 17 patients reporting previous acute injury, 10 had tetanus-prone wounds. Four of six patients who sought medical help for wound management received tetanus toxoid, but none received tetanus immunoglobulins. The most common symptoms were trismus, dysphagia, and muscular rigidity. Specific treatment consisted of active and passive immunization, wound management, parenteral antibiotics, and benzodiazepines, muscle relaxants or neuromuscular blockades for control of spasms and sedation. All adults were admitted to the intensive care unit and an artificial airway was established. Fourteen of them required ventilator support during the illness. Prophylactic tracheostomy was performed within 24 h after arrival in 12 (92%) of 13 patients. Two patients died with an overall mortality rate of 10%. Sequelae were rare in the patients who survived. Because of inclusion of the diphtheria-pertussis-tetanus (DPT) vaccine in the national Children's Vaccine Program and improvement in obstetrical practices and neonatal care in Taiwan, tetanus mainly occurs in people older than 65 years instead of neonates or children. Waning immunity to tetanus in the elderly and poor wound management practices by primary care physicians were contributory factors. PMID- 11045385 TI - Allergic eosinophilic gastroenteritis in a boy with congenital duodenal obstruction. AB - Eosinophilic gastroenteritis (EG) is a rare allergy-related disease, especially in early childhood. We present the case of a 1 year 4 month old boy with congenital duodenal obstruction who developed EG. That diagnosis of EG was made by a series of imaging studies and was confirmed by upper gastrointestinal (UGI) endoscopic biopsy studies which showed significant tissue eosinophilia in both mucosal and submucosal layers. No evidence of parasite segment or ova was found in the stool and biopsy specimen. Specific IgE antibodies to milk were estimated to be 2 + (CAP system). Cow's milk allergy was highly suspected but not confirmed by consecutive elimination and challenge tests since the child was too much suffered to be tested. There was partial response to the 2-week treatment with Alfare (semi-elemental formula) and oral prednisolone 1 mg/kg/day. One month after initial examination, perforation of the stomach occurred and exploratory laparotomy disclosed stenosis of the duodenum. Congenital duodenal obstruction was diagnosed based on operative findings and previous sonographic findings. There has been only one report of EG in an infant with congenital duodenal obstruction. The nature of the relationships among cow's milk allergy as a possible etiologic factor, congenital duodenal obstruction as an predisposing factor and EG involvement at both mucosal and submucosal layers remains unclear. PMID- 11045386 TI - Penicillamine induced lupus-like syndrome: a case report. AB - Several drugs have been suggested to cause lupus-like syndrome. However, penicillamine-induced lupus-like syndrome has only rarely been reported in patients with Wilson's disease. We describe a 6- year-old Taiwanese girl, with a diagnosis of Wilson's disease in November, 1997, who developed lupus-like syndrome 17 months after penicillamine treatment. After treatment with prednisolone and decrease in the dose of penicillamine, her symptoms subsided gradually. This is the first such case reported in a Taiwanese patient. Because the symptoms of drug-induced lupus (DIL) are nonspecific, subjective and variable, the diagnosis of DIL requires awareness of DIL-inducing potential of chronic medication. PMID- 11045387 TI - Plasmapheresis treatment for recurrent focal sclerosis in pediatric renal allografts. AB - Recurrence of focal segmental glomerulosclerosis (FSGS) in pediatric renal allografts is associated with a poor graft survival. This study reports on plasmapheresis for the treatment of recurrent FSGS in pediatric renal transplant recipients. The records of 100 consecutive pediatric (age <21 years) renal transplants were reviewed. Twenty patients had FSGS as the cause of renal failure. Eight of these (40%) had a recurrence (proteinuria >1 g/m2 per day) within 1 month of transplantation. Five of six patients treated with plasmapheresis went into remission (<0.2 g/m2 per day), receiving a total of 42+/ 26 (12-73) sessions, with the mean number of sessions required to achieve a remission being 24+/-17 (8-51). One patient had a second recurrence 1 year following cessation of plasmapheresis and responded to another course of plasmapheresis. The 1 patient who did not respond to plasmapheresis had a delay in initiation of therapy of 42 days. Plasmapheresis initiated within 48 h of recurrence resulted in earlier remissions and improved graft survival among our patients. Plasmapheresis appears to be effective in treating recurrent FSGS following kidney transplantation and should be started as soon as possible. The number of plasmapheresis sessions used to achieve remission should be adjusted according to response rather than adhering to a fixed protocol. PMID- 11045388 TI - Role of nitric oxide in a toxin-induced model of haemolytic uraemic syndrome. AB - The role of nitric oxide in the pathogenesis of glomerular thrombotic microangiopathy was explored using an established rat model in which ricin with or without lipopolysaccharide induced glomerular thrombosis. Ricin alone caused a small rise in the plasma concentration of nitric oxide (control 9.2+/-0.7 microM, ricin 23.3+/-6.3 microM at 7 h). This increase occurred after the development of glomerular thrombosis. Nitric oxide synthase (NOS) activity in the kidney showed no significant change from control values (control 5.66+/-2.7 pmol/min per ml homogenate, ricin 7.52+/-1.8 pmol/min per ml homogenate, total activity). When ricin and lipopolysaccharide were administered together, calcium-independent NOS activity increased whereas calcium-dependent activity decreased (1.22+/-2.6 pmol/min per ml homogenate). The increase in calcium-independent NOS activity correlated with a high plasma concentration of interleukin-1beta in the ricin plus lipopolysaccharide group (4,036.83+/-1,001.5 pg/ml). These data indicate that thrombus formation in a rat model of haemolytic uraemic syndrome is independent of the effects of nitric oxide. PMID- 11045389 TI - Acute renal responses to angiotensin-converting enzyme inhibition in the neonatal pig. AB - Pharmacological interruption or genetic disruption of the renin-angiotensin system before completion of nephrogenesis produces papillary atrophy and an impaired urinary concentrating ability. The mechanisms involved are yet to be elucidated, but renal hypoperfusion and subsequent ischemia, particularly to the immature renal medulla, may be hypothesized. The acute intrarenal responses to angiotensin-converting enzyme (ACE) inhibition in the newborn piglet were thus investigated by means of regional blood flow distribution, renal interstitial hydrostatic pressure (RIHP), and medullary oxygen tension (PO2) in the anesthetised 4- to 5-day-old piglet. Moreover, the calcium antagonist nifedipine and the nitric oxide synthesis inhibitor L-NAME were also given in order to reduce renal blood flow by other means. The drugs were given intravenously in equipotent pressor doses, mimicking intraperitoneal injections in neonatal rats. Enalaprilat (200 microg/kg) reduced mean arterial pressure (MAP) by 14+/-10% (mean+/-SD, P=0.006) and RIHP by 18+/-18% (P=0.001), whereas total renal blood flow and medullary PO2 remained unchanged. In contrast, nifedipine (0.5 mg/kg) reduced MAP and RIHP by 39+/-8% and 38+/-14%, respectively, total and regional blood flows by 30%-60%, and medullary PO, by 46+/-29% (P=0.001). Acute administration of L-NAME (15 mg/kg) increased MAP by 27+/-10% (P=0.0005), whereas RIHP and renal blood flow decreased by 20%-50%, resulting in a reduction of the medullary PO2 by 10+/-12% (P=0.05). We conclude that the renal abnormalities observed after neonatal ACE inhibition are not likely to be caused by renal ischemia. PMID- 11045390 TI - A stepwise approach to the treatment of early onset nephrotic syndrome. AB - We have retrospectively reviewed our single-center experience of the treatment of early onset nephrotic syndrome (NS). From 1991 to 1998, ten children with NS were treated. Kidney biopsy showed focal sclerosis (n=1), diffuse mesangial sclerosis (n=7), and congenital NS of the Finnish type (n=2). Associated conditions included incomplete Drash syndrome (n=1), Galloway-Mowat syndrome (n=1), and severe mental and motor retardation of unknown origin (n=3). From 1991 to 1997, five children with NS were treated. Bilateral nephrectomy (NX) was performed in three, one patient with severe retardation died at 4 years and NX was not performed in one patient who showed satisfactory growth and development. Three of these children were dialyzed and two were successfully transplanted. One patient was transplanted without previous dialysis. From 1997 to 1998, five children were treated with a regimen that included captopril and indomethacin (CAPTO/INDO). CAPTO/ INDO was successful in increasing serum protein in all patients and producing growth and development in four patients. In two patients CAPTO/INDO was successful only after unilateral NX. Our experience indicates that CAPTO/INDO may be a valuable treatment in patients with early onset NS. An individualized stepwise approach including unilateral NX should be considered to achieve optimal results. PMID- 11045391 TI - A randomized prospective crossover trial of amlodipine in pediatric hypertension. AB - Amlodipine has potential advantages in children since it can be dissolved into a liquid preparation and has a long elimination half-life, allowing for once-daily administration. The objective of this study was to compare the efficacy and compliance of amlodipine with that of standard long-acting calcium channel blockers (felodipine or nifedipine) in hypertensive children. A randomized, prospective, crossover study of 11 hypertensive children (9-17 years of age, 10 renal transplant patients) was performed with electronic monitoring of compliance. Each treatment arm was 30 days. No significant differences were observed in mean systolic (SBP) and diastolic blood pressures (DBP) between amlodipine and the other calcium channel blockers. Using 24-h blood pressure monitoring there were no significant differences over each drug treatment period in both mean day-time and night-time SBP and DBP. Patient compliance was similar in both the amlodipine and the nifedipine/felodipine treatment periods. These data suggest that amlodipine is as effective in pediatric nephrology patients as nifedipine and felodipine. Amlodipine may be optimally suited for treatment of young children because at present it is the only calcium channel blocker which can be administered once daily as a liquid preparation. PMID- 11045392 TI - Enalapril and prednisone in children with nephrotic-range proteinuria. AB - The effect of enalapril and low prednisone doses on the urinary protein electrophoretic pattern was studied in 13 pediatric patients with glomerular diseases and steroid-resistant nephrotic syndrome. Enalapril was administered at doses of 0.2-0.6 mg/kg per day for 24-84 months, and prednisone was introduced 2 months later in 11 patients at doses of 30 mg/m2 on alternate days. The urine protein electrophoretic pattern showed a reduction of 80% and 70% in the total protein and albumin, respectively, after enalapril. Total urinary protein decreased from 5.46 to 1.1 g/m2 per day (P<0.001). A marked change from a pattern of non-selective urinary protein loss to an albumin-selective proteinuria was observed. Mean total plasma proteins increased from 4.7 to 5.43 g/dl (P<0.001). Four patients became free of proteinuria 24 months after enalapril was started, but only 2 remained free of proteinuria at 48 months of follow-up. The other 11 patients had persistent albuminuria of between 0.5 and 2.6 g/m2 per day with a selective urinary electrophoretic pattern. No additional decrease was observed after steroids were introduced. A clinical improvement in edema was observed in all children. Three patients developed transient acute renal failure, during the course of an infectious disease; 2 developed peritonitis and 1 pneumopathy. In these patients withdrawal of enalapril was necessary until a complete recovery of renal function was observed. Four patients were hypertensive on admission, achieving normal blood pressure 1 month after enalapril was started. No episodes of systemic arterial hypotension were seen. Creatinine clearance and serum potassium showed no statistically significant change. PMID- 11045393 TI - Hemolytic uremic syndrome associated with Denys-Drash syndrome. AB - The Denys-Drash syndrome is defined by the occurrence of combinations of pseudohermaphroditism, nephrotic syndrome with diffuse mesangial sclerosis, Wilms' tumor, and constitutional mutations in the WT1 suppressor gene. Most patients develop end-stage renal failure. Atypical hemolytic uremic syndrome (HUS) is defined by onset of acute hemolytic anemia with fragmented erythrocytes, thrombocytopenia, and renal failure in the absence of a gastrointestinal prodromal illness of bloody diarrhea. The purpose of this report is to describe the occurrence of features of atypical HUS and Denys-Drash syndrome in two African-American boys aged 13 and 16 months. Each had nephrotic syndrome, diffuse mesangial sclerosis, and WT1 point mutations. Both had grade III hypospadias and undescended testes. They had normal serum creatinine concentrations and hematology a month before presenting with HUS. Stool cultures for Escherichia coli O157:H7 were negative. Each patient has been transplanted with cadaver kidneys without recurrence of HUS. PMID- 11045394 TI - Clinical course and outcome for children with multicystic dysplastic kidneys. AB - The purpose of this study was to evaluate the clinical course and outcome for children with multicystic dysplastic kidney (MCDK) disease and to non-invasively predict which of these patients are at significant risk for developing urinary tract infection (UTI) and renal insufficiency. Patients were divided, on the basis of postnatal physical examination and renal ultrasonography, into simple or complex MCDK. Simple MCDK was defined as unilateral renal dysplasia without additional genitourinary (GU) abnormalities. Complex MCDK included patients with bilateral renal dysplasia or unilateral renal dysplasia with other GU abnormalities. The designation as simple or complex MCDK was independent of reflux, since routine voiding cystourethrography (VCUG) was not performed. The charts of all patients with the diagnosis of MCDK disease seen from August 1995 to March 1999 at Yale University School of Medicine were examined to determine: (1) if UTI had occurred and (2) the level of renal function at last follow-up. Thirty-five patients were evaluated: 28 (80%) patients had unilateral MCDK, 7 (20%) were bilateral, and 14 (40%) had associated GU anomalies. Overall, 21 patients had unilateral MCDK without GU abnormalities (simple MCDK), while 14 had complex MCDK. The final outcome for patients with simple MCDK was quite good, with normal renal function and compensatory hypertrophy of the contralateral kidney in all patients. Although the patients with simple MCDK did not have routine VCUG or prophylactic antibiotics, the development of UTI was infrequent, damage to the contralateral kidney did not occur, and renal function was well preserved. In contrast, patients with bilateral disease or associated GU anomalies had a higher incidence of UTI and progression to renal failure. Complex MCDK was associated with a worse outcome (50% chronic renal insufficiency or failure). PMID- 11045395 TI - Idiopathic hypercalciuria and hyperuricosuria: family prevalence of nephrolithiasis. AB - We studied the prevalence of a history of nephrolithiasis in first- and second degree relatives of 74 children with hypercalciuria (HC), 61 with hyperuricosuria (HU), and 41 with HC plus HU, and in a control population of 261 children with different diseases. Family history of nephrolithiasis was found in 69% of HC, 75% of HU, 78% of HC plus HU, and 22% of control patients. The prevalence was not different among HC, HU, and HC plus HU groups, but was significantly higher in each study group than the control group (P=0.0001). Body mass index >95th percentile was found in only 4.7% of the patients with HC or HC plus HU. Calculi (>3 mm in diameter) were present in 8.9% of the patients with a family history of nephrolithiasis and in 9.4% of those with no family history (P=0.85). Microcalculi (<3 mm in diameter) were found by sonography in 56.6% of the patients with and in 53.3% of those without a family history of nephrolithiasis (P=0.83). Children with HC and/or HU have a strong familial prevalence of nephrolithiasis. Obesity does not seem to affect the association of familial nephrolithiasis and hypercalciuria in children. The presence of nephrolithiasis in families of children with HC and/or HU is not associated with a higher rate of formation of calculi or microcalculi. PMID- 11045396 TI - Pediatric urolithiasis in southern Israel: the role of uricosuria. AB - We describe three cases of severe obstructive uropathy in children under 2 years of age, due to radiolucent renal stones. Metabolic work-up revealed only normouricemic hyperuricosuria (HU) as the single identifiable risk factor for urolithiasis (UL) in these infants. We reviewed records of 66 cases of pediatric UL seen in our service over an 8-year period. UL prevalence was greater for Bedouin than for Jewish children (1.02 vs. 0.13 cases/1,000 inhabitants at risk respectively, P<0.01). HU (>0.6 mg uric acid/dl GFR) was the only biochemical risk factor that differed between Bedouin and Jewish children (mean uric acid excretion index 0.8+/-0.39 vs. 0.55+/-0.26 mg/dl GFR respectively, P<0.05). Bedouin children comprised 85% of patients in the HU group versus 59% in the non hyperuricosuric group (P<0.05). The mean age of onset of UL was 38+/-44 months and 93+/-52 months in the HU and the non-HU group, respectively (P<0.05). The UA excretion index in the HU group was inversely correlated with age (r=0.41, P<0.01) and its slope and constant were different from an age-matched non-UL control population. In conclusion, pediatric UL in southern Israel is predominant in Bedouin toddlers. HU was the only identifiable biochemical risk factor that could explain this difference. PMID- 11045397 TI - Frequency of renal malformations in Turner syndrome: analysis of 82 Turkish children. AB - We evaluated the frequency of renal malformations in relation to nonmosaic 45,X (group A, 45 patients, 54.9%) and mosaic/structural abnormalities of X (group B, 37 patients, 45.1%) in 82 Turkish patients with Turner syndrome (TS). Ultrasonography of the kidneys and collecting system was performed in all patients. Of the 82 patients, 31 had different renal malformations (37.8%). Horse shoe kidney was observed in 9 (29.0%) of the 31 patients, and 17 patients (54.8%) had various collecting system malformations, while 5 (16.2%) had malrotation and other positional abnormalities. The prevalence of renal malformations was significantly higher in group A (51.1%) than group B (21.6%) (2:7.94, P<0.05). Although 8 of the 9 patients with horse-shoe kidney had the 45,X karyotype, collecting system malformations were observed more frequently in group B. Recurrent urinary tract infections (UTIs) were detected during follow-up in 7 patients, and hypertension developed in 3 patients. In patients who had a normal baseline nephrological evaluation, no problem suggesting renal disease developed during follow-up. We conclude that all forms of TS should have routine nephrological screening on diagnosis, since structural malformations of the kidney occur more frequently in nonmosaic 45,X TS, while collecting system malformations are mostly seen in mosaic/structural X forms. Those included in the group for nephrological follow-up had an increased risk for hypertension and/or UTI. PMID- 11045398 TI - Cardiac malformations associated with the congenital nephrotic syndrome. AB - The association of cardiac malformation with the congenital nephrotic syndrome (CNS) has been previously reported in only one family. We report four patients with CNS: three with pulmonary valve stenosis (one requiring valvuloplasty) and one with discrete subaortic stenosis requiring surgical resection. We conclude that the cardiac status of all patients with CNS should be reviewed regularly by a paediatrician, with a low threshold for referral to a cardiologist, as flow murmurs due to chronic anaemia may obscure cardiac pathology. It is important to diagnose any associated cardiac lesions as these may require intervention, and may also predispose to the development of bacterial endocarditis if surgical or dental procedures are undertaken without appropriate antibiotic prophylaxis. PMID- 11045399 TI - Hemolytic uremic syndrome in a child with leukemia and cytomegalovirus infection. AB - A 2-year-old boy had a severe cytomegalovirus (CMV) infection with multi-organ involvement, while on maintenance therapy for acute lymphoblastic leukemia. The patient was treated with intravenous gancyclovir, with a marked improvement in his clinical status, with the exception of a progressive deterioration of the renal function. He also developed hemolytic anemia and thrombocytopenia, suggesting a diagnosis of atypical hemolytic uremic syndrome (HUS). A percutaneous renal biopsy showed lesions consistent with HUS, but no evidence of CMV infection. The patient had a good clinical outcome with no evidence of renal sequelae. We report a rare association of CMV infection and HUS in the pediatric age-group, which suggests a possible cause-effect relationship that deserves further evaluation. PMID- 11045400 TI - New insights into the pathogenesis of renal tubular acidosis--from functional to molecular studies. AB - The diagnosis and classification of renal tubular acidosis (RTA) have traditionally been made on the basis of functional studies. On these grounds, RTA has been separated into three main categories: (1) proximal RTA, or type 2; (2) distal RTA, or type 1; and (3) hyperkalemic RTA, or type 4. In recent years significant advances have been made in our understanding of the subcellular mechanisms involved in renal bicarbonate (HCO3-) and H+ transport. Application of molecular biology techniques has also opened a completely new perspective to the understanding of the pathophysiology of inherited cases of RTA. Mutations in the gene SLC4A4, encoding Na+-HCO3- cotransporter (NBC-1), have been found in proximal RTA with ocular abnormalities; in the gene SLC4A1, encoding Cl(-)-HCO3- exchanger (AE1), in autosomal dominant distal RTA; in the gene ATP6B1, encoding B1 subunit of H+-ATPase, in autosomal recessive distal RTA with sensorineural deafness; and in the gene CA2, encoding carbonic anhydrase II, in autosomal recessive osteopetrosis. Syndromes of aldosterone resistance have been also characterized molecularly and mutations in the gene MLR, encoding mineralocorticoid receptor, and in the genes SNCC1A, SNCC1B, and SCNN1G, encoding subunits of the epithelial Na+ channel, have been found in dominant and recessive forms of pseudohypoaldosteronism type 1, respectively. It can be concluded that, although functional studies are still necessary, a new molecular era in the understanding of disorders of renal acidification has arrived. PMID- 11045401 TI - Retinoids and nephron mass control. AB - Advances in the molecular biology of retinoids have provided evidence that vitamin A profoundly influences the differentiation of the whole embryo. In addition to its well-characterized role in primary body axis and central nervous system formation, vitamin A is also required for the ad hoc development of numerous tissues and organs, including the kidney. This review will focus on the emerging evidence that the development of the urogenital tract depends on retinoids. In order to understand the role of vitamin A during kidney development, the mechanisms and sites of retinoic acid production are presented. In addition, an overview of the molecular targets that may be regulated by retinoic acid is included. Together, these elements support the concept that control of vitamin A homeostasis during renal organogenesis might control nephrogenesis via specific gene expression. The clinical impact of variations in vitamin A status during pregnancy is discussed. PMID- 11045402 TI - Clinical quiz. Molybdenum cofactor deficiency. PMID- 11045403 TI - Necessary practical treatment of cystinuria at night. PMID- 11045404 TI - Historical background to accidental death and disability: the neglected disease of modern society. PMID- 11045405 TI - Effects of prehospital nitroglycerin on hemodynamics and chest pain intensity. AB - OBJECTIVE: To assess the effects of prehospital nitroglycerin (NTG) on vital signs and chest pain intensity. METHODS: A retrospective review of advanced life support (ALS) run sheets was performed in a suburban volunteer emergency medical services (EMS) system receiving 8,000 annual ALS calls. All consecutive patients who were administered NTG by EMS were included. Standardized forms were used to collect data on patient demographics, history, and physical exam. Patients assessed their chest pain (CP) before and after NTG on a verbal numeric scale of 0-10 from least to most severe. The presence of syncope, dysrhythmias, or profound hypotension [loss of peripheral pulses, a systolic blood pressure (SBP) of <90 mm Hg after NTG, or a drop of >100 mm Hg in BP] was noted. Results. One thousand six hundred sixty-two patients received NTG over 18 months, their mean age was 66 years, and 48% were female. Indications for NTG included CP (83%), dyspnea (45%), and congestive heart failure (20%). After NTG administration, the CP score decreased from 6.9 to 4.4 (mean difference = 2.6; 95% CI = 2.4 to 2.8). The CP completely resolved in 10% of the patients. Mean decreases in SBPs and diastolic BPs were 11.8 mm Hg (95% CI = 10.7 to 13.0) and 4.0 mm Hg (95% CI = 2.9 to 5.1). The mean pulse rate increased by 2.7 beats/min (95% CI = 0.6 to 4.9). There were 12 patients with adverse events [0.7% (95% CI = 0.4% to 1.3%)], including profound bradycardia and hypotension (1), transient drop in SBP of 100 mm Hg responding to fluids (6), post-NTG SBP <90 mm Hg (4), and syncope (1). There were no deaths in the prehospital setting. CONCLUSIONS: Use of prehospital NTG appears safe. While NTG reduces CP, most patients have residual pain. PMID- 11045406 TI - The Washington, DC, Youth Curfew: effect on transports of injured youth and homicides. AB - OBJECTIVE: Curfews are implemented to curtail youth violence. Trauma systems and emergency medical services (EMS) may need to prepare for changes in patient volume resulting from local ordinances. This study evaluated the impact of the 1995 Washington, DC, Juvenile Curfew Act on EMS transports of injured youth and on youth homicides. METHODS: A retrospective, comparative cohort study was performed. Transports of injured youth and youth homicides were counted in corresponding months of 1994 and 1995. Cohorts were formed by year, time of day, age group, and mechanism of injury. Year-to-year statistical comparisons of injury proportions were performed using the chi square and Fisher's exact tests. RESULTS: One thousand forty-eight transports were included. No significant difference was observed in transports with curfew implementation. Most assaults on youth occurred outside the curfew time. No effect of the curfew on homicides was detected. Of 67 homicides, only two victims were under the curfew. CONCLUSION: No effect of the curfew on transports for injuries or on homicides was demonstrated. The curfew was not in effect during the period of highest risk. PMID- 11045407 TI - A case series analysis of mass casualty incidents. AB - INTRODUCTION: Mass casualty incidents (MCIs) are infrequent but potentially overwhelming events that can stress the capabilities of even the most organized emergency medical services (EMS) system. The Maryland EMS system has been identified as a pioneer and leader in the field of prehospital emergency care and, as with many states, Maryland's regional preparation for MCIs has been integrated into its overall EMS systems planning. OBJECTIVE: To determine how successful this integration has been by examining a three-year history of response to MCIs in Maryland. METHODS: A three-year case series of MCIs in Maryland was obtained from a Nexis national news publications search. These MCIs were cross-referenced with U.S. postal ZIP codes and the U.S. Census Bureau's ZIP code files. They were then mapped and summary statistics were prepared for analysis. Data obtained through the Maryland Health Services Cost Review Commission for all severely injured patients discharged from Maryland hospitals were obtained over the same three-year period for comparison. RESULTS: Eight MCIs occurred over a three-year period, resulting in a total of 203 injuries. An average of 25.4 +/- 10.7 injuries occurred per MCI. A total of 158 (77.8%) of injuries necessitated ambulance transportation. An average of 3.1 +/- 1.1 hospitals were involved per MCI. CONCLUSIONS: The Maryland EMS system was effective in responding to MCIs ranging in size from 10 to nearly 40 injuries. Analyzing MCIs that reoccur on a year-to-year basis should figure into the planning process for EMS systems. PMID- 11045408 TI - A comparison of biphasic and monophasic shocks for external defibrillation. Physio-Control Biphasic Investigators. AB - BACKGROUND AND OBJECTIVE: The ability of a shock to defibrillate the heart depends on its waveform and energy. Past studies of biphasic truncated exponential (BTE) shocks for external defibrillation focused on low energy levels. This prospective, randomized, double-blind clinical trial compared the first-shock efficacies of 200-joule (J) BTE, 130-J BTE, and 200-J monophasic damped sine wave shocks. METHODS: Ventricular fibrillation (VF) was induced in 115 patients during evaluation of implantable cardioverter-defibrillator function and 39 patients during electrophysiologic evaluation of ventricular arrhythmias. After 19 +/- 10 seconds of VF, a randomized transthoracic shock was administered. Mean first-shock success rates of the three groups were compared using a "Tukey like" statistical test, adjusting for multiple comparisons. Blood pressures and arterial oxygen saturations were measured before VF induction and 30, 90, and 150 seconds after successful defibrillation. RESULTS: First-shock success rates were 61/68 (90%) for 200-J monophasic, 39/39 (100%) for 200-J biphasic, and 39/47 (83%) for 130-J biphasic shocks. The 200-J biphasic shocks were simultaneously superior in first-shock efficacy to both 200-J monophasic and 130-J biphasic shocks (experimentwise error rate, alpha < 0.01). There was no significant difference between the efficacies of 200-J monophasic and 130-J biphasic shocks, nor was there any significant difference between the three groups in hemodynamic parameters after successful shocks. CONCLUSIONS: Biphasic shocks of 200 J provide better first-shock defibrillation efficacy for short-duration VF than 200-J monophasic and 130-J biphasic shocks and thus may allow earlier termination of VF in cardiac arrest patients. PMID- 11045409 TI - Automated external defibrillators in long-term care facilities are cost effective. AB - OBJECTIVE: To assess the cost per life saved of equipping long-term care facilities (LTCFs) with automated external defibrillators (AEDs). METHODS: Outcomes for cardiac arrests within LTCFs were retrieved for 1994 to 1997 from a comprehensive out-of-hospital cardiac arrest registry in a mid-sized U.S. city. The total expense for all LTCFs to obtain and maintain AEDs and to educate and maintain staff skill was estimated for a theoretical four-year period. The cost per life saved to the time of hospital discharge was calculated based on an estimated survival rate of 25% of patients found in ventricular fibrillation (VF) with placement of AEDs in LTCFs. A sensitivity analysis that varied survival rates and costs was conducted. RESULTS: Over four years, there were 160 actual arrests in 43 LTCFs, with a hospital discharge survival rate of 2/160. Twenty of 160 presented to emergency medical services in VF. Training costs for four years were $1,225 per AED. Purchase and maintenance expenses for one AED over four years were $3,941. Placing AEDs in LTCFs would cost $87,837 per life saved if 25% of patients found in VF survived to hospital discharge. Sensitivity analysis using survival rates of 5%, 15%, and 35% established the cost per life saved at $439,184, $146,395, and $62,741, respectively. When costs were calculated at one half and twice the estimated expense, the cost per life saved was $43,918 and $175,674, respectively. CONCLUSION: Placing AEDs in LTCFs is cost-effective at $87,837 per life saved, if a hospital discharge survival rate of 25% of patients in VF can be achieved. PMID- 11045410 TI - Reduction of the call-response interval with ambulance base paging. AB - OBJECTIVE: To determine whether the call-response interval for an emergency medical services (EMS) system would be decreased through the introduction of ambulance base paging. METHODS: The study community included a mixture of urban and rural areas with a total population of approximately 400,000. The EMS system is composed of two ambulance services and one central ambulance communication center with computer-aided dispatching capabilities. Approximately 30,000 calls are responded to yearly by the combined ambulance services. A before-and-after study design was used. In a retrospective review of one ambulance service, there were 224 calls collected in the period before base paging and 200 calls collected in the period after base paging was introduced. In the other ambulance service, there were 571 calls captured in the period before base paging and 515 calls captured in the period after base paging. RESULTS: The call-receipt-to-crew notified interval was reduced from the before period to the after period in both ambulance services: Cambridge--61.8 to 49.8 seconds (p < 0.0001); Kitchener--66.6 to 46.2 seconds (p < 0.0001). The crew-notified-to-vehicle-mobile interval was reduced from the before period to the after period in both ambulance services: Cambridge--91.8 to 73.2 seconds (p < 0.0001); Kitchener-80.4 to 66.0 seconds (p < 0.0001). CONCLUSIONS: The introduction of ambulance base paging reduced components of the call-response interval in this EMS system. Overall, the reduction in time was approximately 30 seconds, which was found to be statistically significant. PMID- 11045411 TI - The strength of specific EMS dispatcher questions for identifying patients with important clinical field findings. AB - INTRODUCTION: There is growing interest in more efficiently matching emergency medical services (EMS) resources to patient need. Emergency medical services dispatchers may be asked to distinguish between callers with an immediate need for EMS and those who may safely use alternative services. New dispatcher protocols are required or existing protocols must be shown to be reliable for this new task. OBJECTIVE: To examine whether answers to currently asked dispatcher questions in one urban center can identify callers with important clinical field findings (ICFFs). METHODS: Audio recordings of EMS dispatcher caller conversations within three nature codes (falls, sick, trauma) were retrospectively reviewed. Specifically scripted "cardinal" questions, asked of all callers, identify what happened, whether the patient is breathing okay, and whether the patient is conscious. "Key" questions are specific to each nature code and further specify patient circumstances. Compliance with protocol and caller answers were documented. Researchers developed a list of ICFFs that, if present on the corresponding EMS record, were judged to justify an immediate EMS response. Logistic regression was used to analyze the relationship between caller answers and the presence of ICFFs. A p-value of 0.10 was used. RESULTS: Of 430 recordings, 383 (89%) were usable. Falls: 103 (26%); trauma: 136 (37%); sick: 144 (37%). The caller was the patient 41 (11%) times. There were 198 (52%) females in the sample. There was no matching EMS record for 96 (25%) cases. An ICFF was determined to be present in 191 (67%) of the 287 recordings with matching EMS data. Compliance across the cardinal and key questions ranged from 62% to 88%. Age alone was suggestive of a patient who may be identified at dispatch as having an ICFF [adjusted OR 1.01 (90% CI: 0.999-1.025), p < 0.10]. No other key or cardinal questions were related to ICFFs. CONCLUSION: Cardinal questions are most often asked. Implied or volunteered information is often relied upon to answer key questions. Key questions for certain nature codes are not answered about one third of the time. Increasing age may suggest a stronger likelihood for an ICFF to be identified at dispatch. Otherwise, in this sample, caller answers to currently asked questions do not appear useful if the goal is to identify at dispatch those without an ICFF. PMID- 11045412 TI - Delays in the EMS response to and the evacuation of patients in high-rise buildings in Singapore. AB - BACKGROUND: Singapore is a highly urbanized and cosmopolitan city situated at the crossroads of Southeast Asia. High-rise buildings and "vertical living" are common, and the city serves as a major business, financial, and industrial hub in the region. More than 80% of the population live in high-rise apartments. This poses unique problems and challenges for emergency ambulance services personnel in the access to and evacuation of patients. OBJECTIVE: To estimate the arrival to-patient contact delay when accessing patients in high-rise buildings and evacuating them to the hospital, compared with accessing patients in ground-level premises. METHODS: This was a prospective study carried out from February 2 to March 1, 1998, for emergency calls from two of the busiest fire stations. The first 150 consecutive cases were enrolled into each of the two groups. Cases of road traffic accidents were excluded because these did not require the crew to get into a building. The times were clocked by one of the paramedics, using a stopwatch. A high-rise building was defined as one where the crew had to ascend at least one flight of stairs. A ground-level building did not involve any stair climbing. We set forth to determine whether the interval between the following was statistically significant when comparing high-rise vs ground-level premises: 1) time when the ambulance arrives at the scene (taken as the time when the driver turns the engine off) and time of arrival at the patient's side; 2) time of leaving the dwelling with the patient and time when the ambulance starts its journey to the hospital (taken as the time when the driver starts the engine). Data analysis was done with the use of SPSS, and the one-tailed unpaired Student's t-test was used for significance testing, with the alpha error rate set at 0.05. Results. One hundred fifty runs were analyzed for each group. The mean delay from arrival to patient contact was 2.49 +/- 0.98 minutes for the high-rise group compared with 1.02 +/- 1.41 minutes for the ground-level group (difference was statistically significant with 95% CI: 1.20, 1.74 minutes; p = 0.0106). The mean delays from the time of leaving the building with the patient to the time when the ambulance turned its engine on to start its journey to the hospital were 3.24 +/- 1.58 minutes and 1.27 +/- 0.71 minutes for the two groups, respectively (difference was statistically significant with 95% CI: 1.68, 2.04 minutes; p = 0.0098). CONCLUSION: There were significant delays present when accessing patients in high-rise buildings and evacuating them to the hospital. Modification to buildings and increasing public awareness and education have been suggested to help minimize these delays. PMID- 11045413 TI - Successful prehospital airway management by EMT-Ds using the combitube. AB - OBJECTIVE: To evaluate the ability to train emergency medical technicians defibrillation (EMT-Ds) to effectively use the Combitube for intubations in the prehospital environment. METHODS: This was an 18-month prospective field study in which EMT-Ds were trained how and in what situations to use the Combitube. Data were then obtained for all patients in whom Combitube insertion was attempted. Indications for use of the Combitube included: unconsciousness without a purposeful response, absence of the gag reflex, apnea or respiratory rate less than 6 breaths/min, age more than 16 years, and height at least 5 feet tall. Contraindications were: obvious signs of death, intact gag reflex, inability to advance the device due to resistance, or known esophageal pathology. Data were entered prospectively from the San Diego County EMS QANet database for prehospital providers. RESULTS: Twenty-two EMT-D provider agencies, involving approximately 500 EMT-Ds, were included as study participants. Combitube insertions were attempted in 195 prehospital patients in cardiorespiratory arrest, with appropriate indication for Combitube use. An overall successful intubation rate (defined as the ability to successfully ventilate) of 79% was observed. Identical success rates for medical and trauma patients were noted. The device was placed in the esophagus 91% of the time. Resistance during insertion was the major reason for unsuccessful Combitube intubations. An overall hospital admission rate of 19% was observed. No complications were reported. CONCLUSION: EMT-Ds can be trained to use the Combitube as a means of establishing an airway in the prehospital setting. Future studies will need to further evaluate its effect on patient outcome. PMID- 11045414 TI - Helicopter emergency medical services roles in disaster operations. AB - Rotor-wing aircraft have previously proven utility in disaster operations, but recent expert reviewers have identified areas of potential improvement in integration of helicopter emergency medical services (HEMS) resources into disaster planning and management. This paper discusses salient points regarding helicopter operations in disaster management, using prior reports regarding rotor wing aircraft utilization as a basis upon which to provide a concise review of HEMS operations in disasters. PMID- 11045415 TI - A statewide study of EMS oversight: medical director characteristics and involvement compared with national guidelines. AB - INTRODUCTION: From their inception, advanced life support (ALS) programs have had oversight by emergency medical services (EMS) medical directors. Position statements about medical direction have been published in the medical literature, initially by the American College of Emergency Physicians (ACEP). Most recently, the National Association of EMS Physicians (NAEMSP) published a position paper to serve as a guideline for the medical directors' tasks, describing qualifications and areas of involvement. OBJECTIVE: To study the baseline status of EMS oversight in Maryland as the position paper was disseminated, to view how local directors meet the published guidelines. METHODS: Twenty-two of the 23 (96%) jurisdictional EMS medical directors (JMDs) in Maryland were interviewed in face to-face meetings. Information was collected about their qualifications and their regular involvement in activities within various EMS subsystems. RESULTS: Sixteen (73%) JMDs are members of ACEP and four (18%) are members of NAEMSP. Six (27%) received EMS medical director training. Three (14%) went through a formal application process for their positions. Activities of relatively high involvement were investigations of variance from protocols (100% of JMDs involved), runsheet review (15 [68%]), liaison duties (20 [91%]), and disaster drills (15 [68%]). Most other subsystems, including dispatch, public health, administration, system evaluation, and quality programs, showed relatively low regular involvement (0-59%). CONCLUSION: In Maryland, significant increases in active physician involvement in EMS are necessary to meet the national job description. These data provide supportive evidence of the need for further commitment by state and local agencies, as well as trained EMS physicians in national societies, to enable all JMDs to reach the goals set forth for their important roles in EMS systems. PMID- 11045416 TI - Rescue from the rubble: urban search & rescue. PMID- 11045417 TI - Early defibrillation. National Association of EMS Physicians Standards and Clinical Practice Committee. PMID- 11045418 TI - Mass gathering medical care. National Association of EMS Physicians Standards and Clinical Practice Committee. PMID- 11045419 TI - Medical direction of interfacility transports: National Association of EMS Physicians Standards and Clinical Practice Committee. PMID- 11045420 TI - EMS Education Agenda for the Future: a systems approach. PMID- 11045421 TI - Sildenafil-nitrate interactions. PMID- 11045422 TI - Modification of the laryngeal tube. PMID- 11045423 TI - Outcomes: giving credit where credit is due. PMID- 11045424 TI - Lentiviral vectors: excellent tools for experimental gene transfer and promising candidates for gene therapy. AB - Lentiviral vectors are tools for gene transfer derived from lentiviruses. From their first application to now they have been strongly developed in design, in biosafety and in their ability of transgene expression into target cells. Primate and non-primate derived lentiviral vectors are now available and with both types of systems a lot of studies tuned to improve their performances in a large number of tissues are ongoing. Here we review the state of the art of lentiviral vector systems discussing their potential for gene therapy. PMID- 11045425 TI - Lentivirus-mediated gene transfer of gp91phox corrects chronic granulomatous disease (CGD) phenotype in human X-CGD cells. AB - BACKGROUND: Chronic granulomatous diseases (CGD) are caused by impaired antimicrobial activity in phagocytes, due to the absence or malfunction of the respiratory burst NADPH oxidase. Two-thirds of the patients have mutations in their X-linked CGD gene encoding gp91phox, the largest subunit of the NADPH oxidase. METHODS: Aimed at gene therapy of X-CGD already at the level of resting pluripotent hematopoietic stem cells, we generated an advanced HIV-1-based vector with self-inactivating (SIN2) features containing the therapeutic gp91phox gene. In this vector an internal cytomegalovirus (CMV) promoter exclusively drives transgene expression. The green fluorescent protein (GFP) served as reporter for evaluation of gene transfer and expression in the human myeloid PLB985 X-CGD cell line. RESULTS: The X-CGD cells were efficiently transduced by the VSV-G pseudotyped lentivirus constructs (up to 74% GFP+ cells at 3 days post transduction). CMV-driven GFP-expression was stable for at least 3 weeks after transduction and persisted after granulocytic differentiation of the target cells. Using the lentivector with the gp91phox transgene, 26% and 48% of the X CGD cells expressed gp91phox at Days 2 and 20 after co-culture with 293T producer cells, respectively. Upon granulocytic differentiation of the transduced X-CGD cells with dimethylformamide (DMF), up to 63% (mean 49%, n = 7) of the cells were found to be functionally reconstituted with mean levels of superoxide production of 31% (n = 7) compared to wild-type PLB985 cells. CONCLUSION: Lentivirus vectors expressing gp91phox are able to at least partially correct human myeloid X-CGD cells. PMID- 11045426 TI - Cardiac functional improvement by a human Bcl-2 transgene in a mouse model of ischemia/reperfusion injury. AB - BACKGROUND: Apoptosis has been shown to contribute to myocardial reperfusion injury. It has been suggested that, in reducing the apoptotic component within the ischemic area at risk, Bcl-2 overexpression could lead to a ventricular function improvement. METHODS: Transgenic mice overexpressing the anti-apoptotic human Bcl-2 cDNA in heart were subjected to a 1-h left coronary artery occlusion followed by a 24-h reperfusion. At the end of the experiment, left ventricular function was assessed by two-dimensional echocardiography. After sacrifice, the area at risk (AR) and the infarct area (IA) were determined by Evans blue and triphenyltetrazolium chloride staining, respectively. The extent of apoptosis was assessed by the TUNEL method. Non-transgenic littermates served as controls. RESULTS: Baseline AR was not different between Bcl-2 transgenic mice and their wild-type littermates. In contrast, left ventricular ejection fraction was significantly improved in the transgenic mice line (61.25 +/- 4.0%) compared to non-transgenic littermates (43.2 +/- 5.0%, p < 0.01). This functional amelioration was correlated with a significant reduction of infarct size in transgenic animals (IA/AR 18.51 +/- 3.4% vs 50.83 +/- 8.4% in non-transgenic littermates). Finally, apoptotic nuclei were less numerous in transgenic mice than in controls as quantified by TUNEL analysis (8.1 +/- 2.2% vs 20.6 +/- 4.4%). CONCLUSIONS: Bcl-2 overexpression is effective in reducing myocardial reperfusion injury and improving heart function. This benefit correlates with a reduction of cardiomyocyte apoptosis. The apoptotic component of ischemia/reperfusion injury could therefore constitute a new therapeutic target in the acute phase of myocardial infarction. PMID- 11045427 TI - NF-kappaB kinetics predetermine TNF-alpha sensitivity of colorectal cancer cells. AB - BACKGROUND: Tumour necrosis factor (TNF)-alpha has considerable anti-tumour activity and may have potential as a treatment for metastatic colorectal cancer. However, TNF-alpha responses in patients and cell lines are variable and TNF alpha treatment is associated with dose limiting clinical toxicity. Activation of NF-kappaB is protective against TNF-alpha induced cell death, and this may explain tumour resistance. METHODS: In order to provide further understanding of determinants of TNF-alpha responses, we studied TNF-alpha induced NF-kappaB activation and variable tumour responses. We analysed the kinetics of TNF-alpha induced NF-kappaB activation in colorectal cancer cells and determined whether it is possible to sensitize colorectal tumour cells to TNF-alpha by modulation of NF kappaB signalling. RESULTS: We demonstrated that sustained NF-kappaB activation exceeding 16 h was observed in HRT18 and SW480 cells and was associated with TNF alpha resistance. In contrast, transient NF-kappaB activation in HCT116 cells was associated with sensitivity to cytotoxic TNF-alpha effects, suggesting that NF kappaB kinetics may have utility as clinical marker of TNF-alpha tumour resistance. Despite variable TNF-alpha responses and NF-kappaB kinetics, all three colorectal cancer cell lines were highly sensitive to treatment with the TNF-related apoptosis-inducing ligand (TRAIL) which induced only transient NF kappaB activation. This further supports the notion of a pre-determined NF-kappaB response influencing receptor-mediated cell death. We also show that stable transfection and adenoviral-mediated expression of IkappaB(A32/36) can be used to confer TNF-alpha sensitivity to colorectal tumour cells previously resistant. CONCLUSIONS: These findings indicate that a combined approach using gene therapy and recombinant TNF-alpha merits further appraisal. Furthermore, the kinetics of the TNF-alpha response could be determined using a 'test-dose' to indicate whether individual patients might benefit from this gene therapy approach. PMID- 11045428 TI - Gene therapy of rat medullary thyroid cancer by naked nitric oxide synthase II DNA injection. AB - BACKGROUND: Nitric oxide (NO), produced by NO synthase II (NOS II), is the main mediator of the tumoricidal action of activated macrophages. In the present study we examined the potential of the NOS II gene as a suicide gene for medullary thyroid cancer (MTC) therapy. METHODS: We orthotopically transplanted rMTC 6-23 cells into the inbred strain of Wag/Rij rats and constructed a plasmid carrying the NOS II gene under the control of the cytomegalovirus (CMV) promoter. RESULTS: Successive injections of tumor cells (Day 0) and naked DNA (Day 2) caused strong inhibition of tumor growth (50%, p < 0.05). Plasmid injection into established tumors (14-day tumors) resulted in the development of large cavities due to tumor cell destruction, with a significant reduction in tumor tissue volume (35%, p < 0.05). Adjacent quiescent tissues were unaffected. Cell death occurred by apoptosis as demonstrated by specific labeling. Macrophages and CD4+ lymphocytes were recruited in the treated tumors. However, tumor-specific T lymphocytes were undetectable in the spleen of treated rats. In control experiments using Lac Z as a reporter gene, expression of beta-galactosidase was detected in only 1% of the tumor cells. CONCLUSIONS: Despite a low gene transfer efficiency, NOS II plasmid produced a strong anti-tumor action resulting from its marked 'bystander' effect mainly due to NO production and diffusion. Therefore the NOS II gene appears to be a promising suicide gene therapy of human cancer. PMID- 11045429 TI - Therapy of peritoneal carcinomatosis from colon cancer with oncolytic adenoviruses. AB - BACKGROUND: The major emphasis on the safety of viral vectors for gene delivery has led to the generally accepted approach of disabling their ability to replicate and thus their potential capacity to spread throughout a tumor by consecutive rounds of infection and lysing neighboring cells. METHODS: In this study we evaluated three herpes simplex virus-1 thymidine kinase (HSV-tk) carrying replication-competent adenoviral vectors with and without the Ad5 E1B 55 kDa gene, wild-type adenovirus type 5 (Ad5wt), and a prototypical replication deficient adenovirus expressing HSV-tk (Ad.TK) for their cytoreductive effects in a peritoneal carcinomatosis model from human HT-29 colon cancer cells in nude mice. RESULTS: The survival of nude mice treated with the replication-defective adenoviral vector Ad.TK was enhanced when followed by GCV. In contrast, administration of GCV diminished the anti-tumor efficacy of the replication competent HSV-tk expressing vectors. However, the intrinsic oncolytic effect of all replication-competent viruses was superior to that of Ad.TK+GCV. Furthermore, the oncolysis of the E1B 55-kDa-positive viruses was significantly greater than that of the E1B 55-kDa-deleted vector. CONCLUSIONS: The more efficient diffusion of viral particles in the peritoneal cavity, when compared to the microenvironment of solid tumors and the virostatic effects of GCV, most likely antagonized the anticipated enhanced cytotoxicity of the replication-competent vectors from the use of its gene directed enzyme prodrug system. Nevertheless, in a clinical setting, the HSV-tk/GCV system allows efficient termination of viral replication in the case of a runaway infection. The results of this study warrant further evaluation of controllable viral replication as a treatment modality for cancer, especially in combination with conventional therapies (e.g. chemotherapy). PMID- 11045430 TI - Evaluation of herpes simplex thymidine kinase mediated gene therapy in experimental pancreatic cancer. AB - BACKGROUND: Despite of recent improvements in the treatment of many malignant diseases, pancreatic ductal adenocarcinoma is still a disease with an extremely poor prognosis with current modes of treatment. Gene therapy has been suggested as a novel approach also against pancreatic cancer. Previous studies have been carried out predominantly with immunodeficient animal models and with tumors growing in environments other than the pancreas. We have attempted to mimic a more clinically relevant situation and investigated suicide gene therapy strategy for experimental pancreatic cancer in animals with an intact immune system. METHODS: We used herpes simplex virus thymidine kinase (HSV-tk) and ganciclovir (GCV) strategy in both in vitro and in vivo settings. RESULTS: In vitro studies demonstrated that retro- as well as adenovirally transduced HSV-tk-positive DSL 6A/C1 rat pancreatic carcinoma cells were sensitive to low concentrations of GCV when compared with parental, nontransduced cells. In addition, a strong bystander effect was observed. In in vivo studies, subcutaneously transplanted HSV-tk positive DSL-6A/C1 cells were killed after GCV treatment, whereas parental, HSV tk-negative cells continued to grow and developed into ductal adenocarcinomas. In in vivo HSV-tk-transduced pancreatic tumors, GCV treatment caused tumor necrosis and the necrosis volume was significantly more extensive when compared with control groups. CONCLUSIONS: HSV-tk gene transfer followed by GCV treatment is efficient in killing pancreatic cancer cells in vitro, in a transduced subcutaneous tumor model, as well as in in vivo transduced pancreatic tumors using an immunocompetent animal model. These results highlight the potential of gene therapy to treat experimental pancreatic cancer. PMID- 11045431 TI - Histidylated polylysine as a synthetic vector for gene transfer into immortalized cystic fibrosis airway surface and airway gland serous cells. AB - BACKGROUND: We recently designed a cationic polymer called histidylated polylysine made of polylysine partially substituted with histidyl residues which become protonated at slightly acidic pH. This polymer is thought to induce the leakage of acidic vesicles containing plasmid/histidylated polylysine complexes. METHODS: and results Here, we have analyzed the ability of histidylated polylysine to transfer reporter or CFTR genes into immortalized cystic fibrosis airway surface epithelial cells (sigmaCFTE29o- cells) and airway gland serous cells (CF-KM4 cells) which are both important targets for cystic fibrosis gene therapy. The luciferase reporter gene expression measured after gene transfer with histidylated polylysine into both cell lines was quite high and similar to that obtained with commercially available vectors. In addition, the level of expression was not dependent on the presence of a membrane disrupting agent such as chloroquine. Histidylated complexes were present in slightly acidic non lysosomal cellular compartments as shown by a cytological approach using biotinylated plasmid, lysosome-specific antibodies and confocal microscopy. Histidylated complexes appeared to be of small size when prepared at low ionic strength and formed aggregates upon increasing the ionic strength. However, aggregate formation was prevented by the addition of 10% fetal bovine serum. Gene transfer efficiency varied with the size of the complexes and decreased when small particles were used. CONCLUSIONS: These results suggest that histidylated polylysine may be an efficient non-viral vector for gene transfer into cystic fibrosis airway surface epithelial cells and airway gland serous cells. PMID- 11045432 TI - HSV-1 infected cell proteins influence tetracycline-regulated transgene expression. AB - BACKGROUND: This study investigates elements of herpes simplex virus type 1 (HSV 1) which influence transgene expression in tetracycline-regulated expression systems. METHODS: Different HSV-1 mutants were used to infect Vero cells that had been transfected with plasmids containing the luciferase gene under the control of tet-off or tet-on tetracycline-regulation systems. RESULTS: The baseline level of luciferase expression was elevated after infection with HSV-1 mutants lacking one or more immediate early genes encoding transactivating factors: ICP27, ICP4 and ICP0. With the tet-off system, not only was baseline expression elevated, but there was a complete loss of induction upon removal of tet when this regulatory system was brought into the cell by infection with helper virus-free amplicon vectors. Elevation of luciferase expression was also observed upon infection with the same HSV-1 mutants following transfection with a plasmid containing only a CMV minimal promoter driving luciferase (pUHC13-3). Only one HSV mutant (14Hdelta3), which bears a disruption in the transactivation domain of VP16 and is deleted for both ICP4 genes, did not increase baseline luciferase expression after transfection of pUHC13-3. The disregulating effects were dependent on virus dose and were not influenced by treatment with interferon (IFN)-alpha, which suppresses viral gene expression. Additional assays involving cotransfection of pUHC13-3 with a plasmid encoding of the HSV-1 transactivating factor ICP4 revealed that ICP4 was the most potent inducer of gene expression from the tetO/CMV minimal promoter. CONCLUSION: These results indicate that proteins encoded in the HSV-1 genome, especially the transactivating immediate early gene products (ICP4, ICP27 and ICP0) and the VP16 tegument protein can activate the tetO/ minimal CMV promoter and thereby interfere with the integrity of tetracycline-regulated transgene expression. PMID- 11045433 TI - Design of an intensive epilepsy monitoring unit. AB - Video-electroencephalography (EEG) telemetry is a crucial component in the comprehensive evaluation of patients with epilepsy. The reasons for patients needing to be monitored fall broadly into three groups: presurgical assessment (36% of our patients), diagnostic assessment (52%), and sleep disorders (12%). Video EEG can be used to differentiate unusual epilepsies from pseudo seizures or other causes of paroxysmal neurological events. The design of a unit depends on the case mix of patients expected to be referred. The key elements to a successful unit are a reliable, flexible, easy-to-use recording system and a team of dedicated, experienced staff, both nursing and technical. The unit at the National Hospital is a six-bed ward with 7 nurses to provide 24-hour coverage, 5 technicians working in shifts, and physics support. A minimum of two staff are on duty at all times. It operates on a five-day week with a throughput of approximately 500 patients per year. It is vital that investigations are performed as efficiently and effectively as possible, and the patient's safety and wellbeing is paramount at all times. Drug reduction is likely to be used to precipitate seizures, especially in those being considered for epilepsy surgery, and this poses a risk of provoked secondary generalized seizures. Continuous supervision of patients, and the ability to respond rapidly to a seizure, are therefore essential. We adopt a standardized easy-to-follow drug-reduction protocol, similar to that used by other centers. PMID- 11045434 TI - Advances in telecommunications concerning epilepsy. AB - Telemedicine is a rapidly expanding discipline. Looking back on a long tradition of telemetric measurement and transmission of brain electrical data, one might ask how relevant telecommunication techniques have become for clinical work in epileptology. In principle, communication can be either between patient and doctor or between doctor and doctor. The former includes telephone reports on frequency and severity of seizures and on mental and physical state. Because of the special problems of patients with epilepsy, the need for traveling should be minimized. To maintain close contact, telemetric transmission of electroencephalograms (EEG), seizure video-registration, and monitoring of anti epileptic drug (AED) blood levels from home are desirable, but the technical tools now available are not of sufficiently high performance for these applications. However, physicians and medical units can communicate using high rate data transfer. There are major problems with this technology. Transfer rates using the internet are not fixed. Moreover, using six digital telephone lines simultaneously, good-quality transmission of a 3-min video may take more than 1 h, restricting interactivity. In contrast to imaging (e.g., magnetic resonance imaging), there is no generally accepted protocol for handling EEG/electrocorticography data. There are also legal problems relating to the responsibility for failures and safety of personal or medical data. Nevertheless, the expenses of installation and use of such equipment appear justified by the expected benefits, for the patients (avoiding travel and multiple investigations, checking diagnosis and therapy, amending quality of life), for the physician (sparing equipment and manpower, accelerating and improving diagnosis), for the epilepsy center (increased database for expert systems, improved logistic and data storage) and, finally, for insurance providers (reduced overall costs). When the neurosurgical procedure is remote from the place of presurgical evaluation, telecommunication should be obligatory. PMID- 11045435 TI - New insights into the clinical management of partial epilepsies. AB - The diagnosis, treatment, and prognosis of seizure disorders depend on the correct identification of epileptic syndromes. Partial epilepsies are heterogeneous and can be divided into idiopathic, cryptogenic, and symptomatic epilepsies. The most common of the idiopathic localization-related epilepsies is benign epilepsy with rolandic or centrotemporal spikes (BECTS). Seizures remain rare and the use of antiepileptic drug (AED) treatment in all patients does not appear justified. Children who present with some of the electroclinical characteristics of BECTS may also display severe unusual neurologic, neuropsychological, or atypical symptoms. In some cases, carbamazepine has been implicated as a triggering factor. Primary reading epilepsy and idiopathic occipital lobe epilepsies with photosensitivity are examples of an overlap between idiopathic localization-related and generalized epilepsies and respond well to sodium valproate. Autosomal dominant nocturnal frontal lobe epilepsy and benign familial infantile convulsions are recently described syndromes, differing in several ways from classical idiopathic localization-related epileptic syndromes. In cryptogenic or symptomatic epilepsy, the topography of the epileptogenic zone might influence drug efficacy. An individualized approach to AED selection, tailored to each patient's needs, should be used. Resistance of seizures to antiepileptic therapy may be due to diagnostic and/or treatment error or may be the result of noncompliance. Increasing the dosage, discontinuation or replacement of a drug, or addition of a second drug is indicated in truly resistant cases. The use of more than two AEDs rarely optimizes seizure control, and in some cases reduction of treatment may improve seizure control while lessening side effects. EEG-video assessment of patients with refractory epilepsy is important. Indications for and timing of epilepsy surgery should be reconsidered. Surgical therapy should probably be used more often and earlier than it is at present. PMID- 11045436 TI - Clinical ictal symptomatology and anatomical lesions: their relationships in severe partial epilepsy. AB - High-resolution imaging techniques can demonstrate anatomic alterations in most patients identified as candidates for surgical treatment of their partial epilepsy. The demonstration of an anatomic lesion is only one step in the presurgical diagnostic procedure, which includes video-EEG and, when necessary, video-stereo-EEG recordings of seizures. A review of the literature shows that the simple removal of the magnetic resonance imaging (MRI)-evident lesion ("lesion-ectomy") reduces but does not completely suppress seizures in a large percentage of patients, especially those with neuronal migration disorders. This phenomenon could, at least in part, be explained by preliminary data (in 33 patients) showing that less than 20% of seizures correspond to a well-localized, intralesional discharge in about 40% of stereo-EEG-investigated patients with at least one intralesional electrode. The authors illustrate some anatomo electroclinical examples of intraindividual variability of the ictal symptomatology, raising the problem of the decision about the extent of the surgical removal. Recent histologic and immunohistochemical studies have demonstrated several kinds of structural alterations in the stereo-EEG-defined epileptogenic zone, not always overlapping with the MRI-visible lesion. This aspect can further explain some failures of MRI-guided lesionectomies. That relationships between "lesions" and epileptogenic zones may be variable is also suggested by reports of patients who present with multiple lesions (i.e., cavernous angiomas, Bourneville syndrome) and are cured by removal of only one of them. PMID- 11045437 TI - Cost minimization analysis of antiepileptic drugs in newly diagnosed epilepsy in 12 European countries. AB - A recent United Kingdom cost minimization analysis (CMA) of four antiepileptic drugs (AEDs) used to treat newly diagnosed adult epilepsy demonstrated that a new drug, lamotrigine (LTG), incurred higher costs than carbamazepine (CBZ), phenytoin (PHT), and valproate (VPA), whose costs were similar. This analysis took account of each drug's side-effect and tolerability profile. The present analysis investigated the costs of treatment with LTG, CBZ, PHT, and VPA in 12 European countries. Data were derived from published sources and from a panel of locally based experts. When no published data were available, estimates were obtained using expert opinion by a consensus method. These data were incorporated into a treatment pathway model, which considered the treatment of patients during the first 12 months after diagnosis. The primary outcome considered was seizure freedom. Randomized controlled trials demonstrate that the drugs considered are equally effective in terms of their ability to achieve seizure freedom, and thus the most appropriate form of economic evaluation is a CMA. These trials provided data on the incidence of side effects, dosages, and retention rates. The economic perspective taken was that of society as a whole and the analysis was calculated on an "intent-to-treat" basis. Only direct medical costs were considered. In each country considered, LTG was twofold to threefold more expensive than the other drugs considered. A sensitivity analysis demonstrated that varying each of the assumptions (range defined by expert panels) did not significantly alter the results obtained. PMID- 11045438 TI - Scientific contribution by Prof. Tsuda. PMID- 11045439 TI - Recent bioorganic studies on rhodopsin and visual transduction. AB - Rhodopsin, the pigment responsible for vision in animals, insect and fish is a typical G protein (guanyl-nucleotide binding protein) consisting of seven transmembrane alpha helices and their interconnecting extramembrane loops. In the case of bovine rhodopsin, the best studied of the visual pigments, the chromophore is 11-cis retinal attached to the terminal amino group of Lys296 through a protonated Schiff base linkage. Photoaffinity labeling with a 3-diazo-4 oxo-retinoid shows that C-3 of the ionone ring moiety is close to Trp265 in helix F (VI) in dark inactivated rhodopsin. Irradiation causes a cis to trans isomerization of the 11-cis double bond giving rise to the highly strained intermediate bathorhodopsin. This undergoes a series of thermal relaxation through lumi-, meta-I and meta-II intermediates after which the retinal chromophore is expelled from the opsin binding pocket. Photoaffinity labeling performed with 3-diazo-4-oxoretinal at -196 degrees C for batho-, -80 degrees C for lumi-, -40 degrees C for meta-I, and 0 degrees C for meta-II rhodopsin showed that in bathorhodopsin the ring is still close to Trp265. However, in lumi-, meta I and meta-II intermediates crosslinking occurs unexpectedly at A169 in helix D (IV). This shows that large movements in the helical arrangements and a flip over of the ring moiety accompanies the transduction (or bleaching) process. These changes in retinal/opsin interactions are necessarily accompanied by movements of the extramembrane loops, which in turn lead to activation of the G protein residing in the cytoplasmic side. Of the numerous G protein coupled receptors, this is the first time that the outline of transduction pathway has been clarified. PMID- 11045440 TI - A new gluco indole alkaloid, 3, 4-dehydro-5-carboxystrictosidine, from Peruvian Una de Gato (Uncaria tomentosa). AB - A new gluco indole alkaloid, 3,4-dehydro-5-carboxystrictosidine, was obtained from Peruvian Una de Gato (Cat's Claw, original plant: Uncaria tomentosa) together with two known gluco indole alkaloids. This compound was the first example of isolation of a gluco monoterpenoid indole alkaloid having a 3,4 dihydro-beta-carboline ring system from nature. A characteristic feature of the compound was the quick replacement of the methylene hydrogens on C-14 with deuterium that was observed when it was dissolved in CD3OD. We demonstrated a similar proton-deuterium exchange on a model compound, 1-methyl-3,4-dihydro-gamma carboline, in CD3OD solution. PMID- 11045441 TI - Ardisimamillosides C-F, four new triterpenoid saponins from Ardisia mamillata. AB - Four new triterpenoid saponins, ardisimamilloside C (1), 3-O-[alpha-L rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->4)-[beta -D-glucopyranosyl-(1- >2)]-alpha-L-arabinopyranosyl]-3beta,16al pha,28,30-tetrahydroxy-olean-12-en, ardisimamilloside D (2), 3-O-?alpha-L-rhamnopyranosyl-(1-->2)-beta-D glucopyranosyl-(1-->4)-[beta -D-glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosyl] 3beta,15al pha,28,30-tetrahydroxy-olean-12-en, ardisimamilloside E (3), 3-O [alpha-L-rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->4)-[beta -D glucopyranosyl-(1-->2)]-alpha-L-arabinopyranosl]-13beta,2 8-epoxy 3beta,16alpha,29-oleananetriol, and ardisimamilloside F (4), 3-O-[alpha-L rhamnopyranosyl-(1-->2)-beta-D-glucopyranosyl-(1-->4)-[beta -D-glucopyranosyl-(1- >2)]-alpha-L-arabinopyranosyl]-3beta,16al pha-dihydroxy-13beta,28-epoxy-oleanan 30-oic acid were isolated from the roots of Ardisia mamillata Hance. Structure assignments were established on the basis of highresolution (HR)-FAB-MS, 1H-, 13C , and two-dimensional (2D)-NMR spectra, and on the chemical evidence. PMID- 11045442 TI - Five lanostane triterpenoids and three saponins from the fruit body of Laetiporus versisporus. AB - Five lanostane triterpenes, named versisponic acids A-E and three lanostanoid glycosides, laetiposides E-G, were isolated from the fruit bodies of Laetiporus versisporus. Their structures were established by extensive NMR experiments and chemical methods. PMID- 11045443 TI - A new pentanorlanostane derivative, cladosporide A, as a characteristic antifungal agent against Aspergillus fumigatus, isolated from Cladosporium sp. AB - In the course of searching for new antifungal agents, a new pentanorlanostane derivative, cladosporide A (1), was isolated along with ergosterol, ergosterol peroxide and 23,24,25,26,27-pentanorlanost-8-ene-3beta,22-diol (2) from Cladosporium sp. as a characteristic antifungal agent against the human pathogenic filamentous fungus Aspergillus fumigatus. The structure of 1 was established as 3beta,22-dihydroxy-23,24,25,26,27-pentanorlanostane-29-al by spectroscopic and chemical investigation and X-ray crystallographic analysis. Inhibitory activity against A. fumigatus (IC80 0.5-4.0 microg/ml) was observed for cladosporide A (1), but no activity was observed against pathogenic yeasts, Candida albicans and Cryptococcus neoformans, and other pathogenic filamentous fungi, Aspergillus niger and A. flavus. The 4beta-aldehyde residue in 1 might be essential for the antifungal activity, since 23,24,25,26,27-pentanorlanost-8-ene 3beta,22-diol (2) showed no inhibition against the above four fungi. PMID- 11045444 TI - Oleanane acid from Myrica cerifera. AB - From the twigs of Myrica cerifera L. (Myricaceae), a new oleanane triterpenic acid named myrica acid was isolated along with myricalactone and several other known constituents. The structure of the acid was determined as 3beta-hydroxy-1 oxoolean-11,13(18)-dien-28-oic acid on the basis of chemical and spectral evidence. PMID- 11045445 TI - Medicinal foodstuffs. XX. Vasorelaxant active constituents from the roots of Angelica furcijuga Kitagawa: structures of hyuganins A, B, C, and D. AB - From the methanolic extract with vasorelaxant activity obtained from Angelica furcijuga Kitagawa, four new khellactone-type coumarins, hyuganins A, B, C, and D, were isolated together with twelve known coumarins, two known acetylenic compounds, and a known lignan. The structures of hyuganins A, B, C, and D were determined on the basis of chemical and physicochemical evidence. Nine principal coumarins (hyuganin A, anomalin, pteryxin, isopteryxin, isoepoxypteryxin, praerosides II and IV, apiosylskimmin, (R)-peucedanol 7-O-beta-D glucopyranoside), two acetylenic compounds [(-)-falcarinol and falcarindioll, and related compounds were examined for inhibitory activities on high concentration of K+ (High K+)- and dl-norepinephrine (NE)-induced contractions. The results indicate that the 3'- and 4'-acyl groups of khellactone-type coumarins are essential for the inhibitory activity on the contractions by High K+. Hyuganin A and anomalin showed inhibitory effects on High K+-induced contraction, but not on NE-induced contraction. Other active coumarins (pteryxin, isopteryxin, isoepoxypteryxin) and an acetylenic compound (falcarindiol) non-selectively inhibited both contractions by High K+ and NE. PMID- 11045446 TI - Immunomodulatory constituents from an ascomycete, Emericella aurantio-brunnea. AB - Fractionation monitored by the immunomodulatory activity of the AcOEt extract of an Ascomycete, Emericella aurantio-brunnea, afforded two known fungal sesterterpenes, variecolin (1) and variecolactone (2), two new variecolin congeners named variecoacetals A (3) and B (4), and a new sesquiterpenetriol diester named emeremophiline (5), as the immunosuppressive constituents of this fungus. The absolute configuration of 1, which was previously not determined, was determined to be (2S,3S,6R,10S,11R,14S,15R,16S) from the NMR spectral data of the (6R,7R)-dimethyl-1,3,5-trioxacycloheptyl derivative of 1 (7). The absolute configurations of the other variecolin congeners, 2-4, and variecolol (6) are also proposed from biosynthetic considerations. PMID- 11045447 TI - Argadin, a new chitinase inhibitor, produced by Clonostachys sp. FO-7314. AB - A new chitinase inhibitor, designated as argadin (1), was isolated from the cultured broth of a fungal strain FO-7314. The strain was identified as Clonostachys sp. from the morphological characteristics. Argadin was purified from the cultured mycelium by a combination of cation exchange, adsorption and gel filtration chromatographic methods. The structure of argadin was elucidated as cyclo(Nomega-acetyl-L-arginyl-D-prolyl-homoseryl-histidyl-L- 2-aminoadipyl) in which homoseryl gamma-methylene bonded to histidyl alpha-amino residue. The IC50 value of argadin against Lucilia cuprina (blowfly) chitinase was 150 nM at 37 degrees C and 3.4 nM at 20 degrees C. Argadin arrested the moult of cockroach larvae upon injection into the ventral abdominal part. PMID- 11045448 TI - Colopsinols D and E, new polyhydroxyl linear carbon chain compounds from marine dinoflagellate Amphidinium sp. AB - Colopsinols D (1) and E (2), two new polyhydroxyl linear carbon chain compounds, have been isolated from the cultured marine dinoflagellate Amphidinium sp. The structures of 1 and 2 were elucidated on the basis of two-dimensional NMR and FAB MS/MS data. PMID- 11045449 TI - Isolation and characterization of bioactive metabolites from marine-derived filamentous fungi collected from tropical and sub-tropical coral reefs. AB - Two new compounds, paecilospirone (1) and phomopsidin (2), and seven known compounds, chaetoglobosin A (3), griseofulvin (4), fusarielin A (5), fusapyrone (6), deoxyfusapyrone (7), and verrucarins J (8) and L acetate (9), have been isolated and characterized from marine-derived fungi collected in tropical and sub-tropical coral reef environments. The utility of marine-derived fungi as a source of bioactive secondary metabolites is discussed. PMID- 11045450 TI - Biochemical and partial molecular characterization of bitter and sweet forms of Lupinus angustifolius, an experimental model for study of molecular regulation of quinolizidine alkaloid biosynthesis. AB - The bitter and sweet forms of a plant species differing with alkaloid contents may provide a model system for investigation of alkaloid biosynthesis at a molecular level. The pattern and concentration of quinolizidine alkaloids were determined by capillary GC-MS in bitter and sweet plants of Lupinus angustifolius. Bitter plant contained lupanine, 13alpha-hydroxylupanine, angustifoline, alpha-isolupanine, tetrahydrorhombifoline, and ester-derivatives of 13alpha-hydroxylupanine. In contrast, no alkaloid was detected in sweet plant. The enzymatic activity of acyltransferase for formation of 13alpha tigloyloxylupanine was similar or even higher in the cell-free extracts of sweet plant than that in bitter plant. These results suggest that the biosynthetic step(s) of ring closure forming the initial cyclic alkaloid, lupanine, from cadaverine is presumably blocked in sweet plant, and that the later steps for modification of the cyclized alkaloids are not altered. We hypothesized that the gene(s) encoding enzyme(s) for ring-closure step might be repressed in sweet plant, and that the expression might take place only in bitter plant. To isolate the genes specifically expressed in bitter plant, cDNA-amplified fragment length polymorphism (cDNA-AFLP) analysis was carried out. However, no bitter-specific gene was isolated, suggesting that alkaloid biosynthesis in sweet plant may be down-regulated at a post-transcriptional level. PMID- 11045451 TI - Determination of alpha-tocopherol and alpha-tocopherylquinone in rat tissues and plasma by high-performance liquid chromatography with electrochemical detection. AB - Alpha-tocopherol and alpha-tocopherylquinone in rat tissues and plasma were determined simultaneously by using high-performance liquid chromatography electrochemical detection (HPLC-ED) with dual electrodes in the series mode. Biological samples were saponified in the presence of a mixture of butylated hydroxytoluene, ascorbic acid, and pyrogallol and then extracted with hexane. The compounds were separated on a C18 column using a mobile phase containing 95% methanol and 0.05 M sodium perchlorate as the supporting electrolyte. After HPLC separation, alpha-tocopherylquinone was first reduced at an upstream electrode at -500 mV Both alpha-tocopherol and the reduction product of alpha tocopherylquinone were then oxidized downstream at +600 mV. Only the downstream electrode current was monitored for the determination. Linearity of the standard curves was obtained over the range 5-30 pmol for alpha-tocopherol and alpha tocopherylquinone. Minimum detectable quantities (S/N of 3) were 0.25 pmol for alpha-tocopherol and 0.31 pmol for alpha-tocopherylquinone. The method was applied to analysis of the contents of alpha-tocopherol and alpha tocopherylquinone in rat tissues and plasma. By hyperoxia, the content of alpha tocopherol was decreased remarkably in lung, and in contrast, the contents of alpha-tocopherylquinone were increased in all tissues studied with the exception of plasma, though the content of alpha-tocopherylquinone in normal rats is quite small. The technique is particularly useful in the quantitation of the oxidation of alpha-tocopherol in biological samples. PMID- 11045452 TI - Antioxidant activity of eugenol and related monomeric and dimeric compounds. AB - Since the inhibitory effect of eugenol (a), which was isolated as an antioxidative component from plant, Caryopylli flos, on lipid peroxidation was less than that of alpha-tocopherol, we synthesized the eugenol-related compounds dieugenol (b), tetrahydrodieugenol (c), and dihydroeugenol (d), to find new strong antioxidants and assessed them for their inhibitory effect on lipid peroxidation and scavenging ability for superoxide and hydroxyl radicals. The antioxidative activities were in the order: (b)>(c)>(d)>(a) for the thiobarbituric acid reactive substance (TBARS) formation. These results suggest that the dimerized compounds have higher antioxidant activities than that of the monomers. Electron spin resonance (ESR) spin trapping experiments revealed that eugenol and its dimer, having allyl groups in the structure, scavenged superoxide, and that only eugenol trapped hydroxyl radicals under the conditions used. These finding suggest that eugenol and dieugenol have a different mechanism of antioxidation, i.e. eugenol may inhibit lipid peroxidation at the level of initiation, however, the related dimeric compounds may inhibit lipid peroxidation at the level of propagation of free radical chain reaction like alpha-tocopherol. PMID- 11045453 TI - Synthesis of NG-061 and its analogs, and their biological evaluation as an enhancer of nerve growth factor. AB - A novel potentiator of nerve growth factor (NGF), NG-061, which had been isolated from the fermentation broth of Penicillium minioluteum F-4627, was synthesized from methoxybenzoquinone and phenylacetylhydrazine in a single step. A series of acyl hydrazone derivatives were also synthesized and their potentiator activity of neurotrophic effect of NGF on neurite outgrowth was evaluated by assay with a rat pheochromocytoma cell line PC12. PMID- 11045454 TI - Synthesis of 15alpha-fluoro-24,25-dihydrolanosterol as a potential inhibitor and/or mechanistic probe for lanosterol 14alpha-demethylase. AB - As a potential inhibitor and/or mechanistic probe for lanosterol 14alpha demethylase, 15alpha-fluoro-24,25-dihydrolanosterol was prepared by fluorination of 15alpha-hydroxy-24,25-dihydrolanost-7-en-3beta-yl benzoate with diethylaminosulfur trifluoride, followed by hydrogen chloride-catalyzed isomerization of the delta7 to delta8 and reductive cleavage of the benzoate. PMID- 11045455 TI - Synthesis of 3-epi-6,7-dideoxyxestobergsterol A. AB - 3-Epi-6,7-dideoxyxestobergsterol A (2), an analogue of xestobergsterol A, has been synthesized from dehydroepiandrosterone (3) in 15 steps. The key synthetic intermediate, 15beta,16alpha-dioxypregn-17(20)E-ene derivative 8, was prepared from the corresponding 15beta,16beta-epoxide 6 by treating with acetic acid and titanium tetraisopropoxide. The 23-oxo side chain was constructed stereoselectively by orthoester Claisen rearrangement of 8 followed by introduction of an isobutyl group. Basic treatment of the 15,23-diketone 12 followed by deprotection gave the title compound 2. PMID- 11045456 TI - Synthesis of (10Z)- and (10E)-19-fluoro-1alpha,25-dihydroxyvitamin D3: compounds to probe vitamin D conformation in receptor complex by 19F-NMR. AB - To study the interaction of vitamin D with its receptor by 19F-NMR, (5Z,10Z)- and (5Z,10E)-19-fluoro-1alpha,25-dihydroxyvitamin D3 were synthesized starting from vitamin D2 via electrophilic fluorination of vitamin D-SO2 adducts as the key step. Regio- and stereoselective electrophilic fluorination at C(19) of vitamin D SO2 adducts was achieved under the conditions using (PhSO2)2NF and bulky bases. The stereochemistry of the addition and elimination of SO2 of various vitamin D derivatives was studied in detail. SO2 causes Z-E isomerization of the 5,6-double bond of vitamin D and adds to the resulting (5E)-isomer from the sterically less hindered side opposite to the substituent at C(1). Elimination of SO2 from 19 substituted vitamin D-SO2 adducts proceeded exclusively in a suprafacial manner with respect to the diene part under either thermal or reductive conditions. Dye sensitized photochemical isomerization of 19-fluorovitamin D derivatives was studied in detail. The rapid isomerization at the 5,6-double bond was followed by the slow isomerization at the 10,19-double bond to yield the (5E,10Z)-isomer (by nomenclature of the 1-OH derivatives) as the major product. (10Z)- and (10E)-19 Fluorovitamin Ds were also interconverted thermally probably via the corresponding previtamin D by 1,7-sigmatropic isomerization. PMID- 11045457 TI - Alpha-glucosidase inhibitors with a phthalimide skeleton: structure-activity relationship study. AB - Alpha-glucosidase inhibitors with a phthalimide skeleton were prepared. Structure activity relationship studies indicated a critical role for the hydrophobicity of the substituent at the nitrogen atom of the phthalimide skeleton. Introduction of electron-withdrawing groups, including a nitro group and chlorine, influenced the activity. Optimization studies led us to design 4,5,6,7-tetrachloro-N phenylphthalimide (CPOP) and its N-phenylalkyl derivatives. CP0P and 4,5,6,7 tetrachloro-N-(4-phenylbutyl)phthalimide (CP4P) proved to be more potent alpha glucosidase inhibitors than the known inhibitor 1-deoxynojirimycin. PMID- 11045458 TI - Synthesis and properties of novel bifunctional nitrosamines with omega chloroalkyl groups. AB - Novel N-nitroso-N-(acetoxymethyl)-omega-chloroalkylamines were synthesized and their chemical and biological properties were evaluated. The nitrosamines were expected to decompose through omega-chloroalkyldiazohydroxides in aqueous solution, and then to alkylate various cellular macromolecules. N-Nitroso-N (acetoxymethyl)-2-chloroethylamine rapidly decomposed in aqueous solution, and the reaction rate was apparently independent of the pH of the solution. On the other hand, the rate of decomposition of chloropropyl and chlorobutyl homologs was pH-dependent, and increased in alkaline solution. When mutagenicity was assayed in Salmonella typhimurium TA1535 and TA92 for preliminary evaluation, all three compounds were directly mutagenic. The mutagenicity in Salmonella typhimurium TA1535, which can detect base-pair change mutation, clearly showed that these compounds induced DNA alkylation in vivo. The increase of alkyl chain length in chloroalkyl compounds increased the mutagenic activity, and the activities were stronger than those of the corresponding simple alpha-acetoxy nitrosamines lacking a chloro group, N-nitroso-N-(acetoxymethyl)alkylamines. Furthermore, the positive result in TA92 suggested that chlorinated nitrosamines cross-linked DNA like antitumor chloroethylnitrosoureas and that they are expected to be new lead compounds for antitumor agents. PMID- 11045459 TI - Retinoidal pyrimidinecarboxylic acids. Unexpected diaza-substituent effects in retinobenzoic acids. AB - Several pyridine- and pyrimidine-carboxylic acids were synthesized as ligand candidates for retinoid nuclear receptors, retinoic acid receptors (RARs) and retinoic X receptors (RXRs). Although the pyridine derivatives, 6-[(5,6,7,8 tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)carbamoyl]pyri dine-3-carboxylic acid (2b) and 6-[(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2 naphthalenyl)carboxamido]py ridine-3-carboxylic acid (5b) are more potent than the corresponding benzoic acid-type retinoids, Am80 (2a) and Am580 (5a), the replacement of the benzene ring of Am580 (5a), Am555 (6a), or Am55 (7a) with a pyrimidine ring caused loss of the retinoidal activity both in HL-60 cell differentiation assay and in RAR transactivation assay using COS-1 cells. On the other hand, pyrimidine analogs (PA series, 10 and 11) of potent RXR agonists (retinoid synergists) with a diphenylamine skeleton (DA series, 8 and 9) exhibited potent retinoid synergistic activity in HL-60 cell differentiation assay and activated RXRs. Among the synthesized compounds, 2-[N-n-propyl-N (5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)a mino]pyrimidine-5 carboxylic acid (PA013, 10e) is most active retinoid synergist in HL-60 assay. PMID- 11045460 TI - Superoxide dismutase activity of iron(II)TPEN complex and its derivatives. AB - Superoxide is involved in the pathogenesis of various diseases, such as inflammation, ischemia-reperfusion injury and carcinogenesis. Superoxide dismutases (SODs) catalyze the disproportionation reaction of superoxide to produce oxygen and hydrogen peroxide, and can protect living cells against the toxicity of free radicals derived from oxygen. Thus, SODs and their functional mimics have potential value as pharmaceuticals. We have previously reported that Fe(II)tetrakis-N,N,N',N'-(2-pyridylmethyl)ethylenediamine (Fe(II)TPEN) has an excellent SOD activity (IC50 = 0.5 microM) among many iron complexes examined (J. Biol. Chem., 264, 9243-9249 (1989)). Fe(II)TPEN can act like native SOD in living cells, and protect Escherichia coli cells from free radical toxicity caused by paraquat. In order to develop more effective SOD functional mimics, we synthesized Fe(II)TPEN derivatives with electron-donating or electron-withdrawing groups at the 4-position of all pyridines of TPEN, and measured the SOD activities and the redox potentials of these complexes. Fe(II) tetrakis-N,N,N',N' (4-methoxy-2-pyridylmethyl)ethylenediamine (Fe(II)(4MeO)4TPEN) had the highest SOD activity (IC50 = 0.1 microM) among these iron-based SOD mimics. In addition, a good correlation was found between the redox potential and the SOD activity of 15 Fe(II) complexes, including iron-based SOD mimics reported in the previous paper (J. Organometal. Chem., in press). Iron-based SOD mimics may be clinically applicable, because these complexes are generally tissue-permeable and show low toxicity. Therefore our findings should be significant for the development of clinically useful SOD mimics. PMID- 11045461 TI - Lipase-catalyzed asymmetric desymmetrization of prochiral 2,2-disubstituted 1,3 propanediols using 1-ethoxyvinyl benzoate. AB - The lipase-catalyzed asymmetric desymmetrization of the prochiral 2,2 disubstituted 1,3-propanediols was studied using various types of 1-ethoxyvinyl esters (1a-i). Although 1a-e with aliphatic acyl groups were not sufficient, use of the benzoate (1f) in combination with Candida rugosa lipases converted acyclic diols (2, 6) and cyclic diols (11-14) to the optically active compounds (3f, 7f, 15f-18f), bearing a quaternary carbon center, with moderate-to-high optical yields. These products were fairly stable against racemization under acidic conditions. PMID- 11045462 TI - Cleavage of S-S bond by nitric oxide (NO) in the presence of oxygen: a disproportionation reaction of two disulfides. AB - Disulfide bond was cleaved by a catalytic amount of nitric oxide in the presence of oxygen, which was confirmed by experiments employing two symmetrical disulfides. The reaction resulted in the formation of unsymmetrical disulfides in nearly 50% yields. The steric hindrance of alkyl disulfide slowed the reaction rate, and an electron-donating group on the aryl disulfide promoted the reaction. The substituent and S-nitrosothiol effects suggested that the reaction was initialized with an oxidative process by NO+. PMID- 11045464 TI - Hydroxylation of nitrated naphthalenes with KO2/crown ether. AB - Superoxide radical anion (O2*-), generated by KO2/crown ether, is effective for hydroxylation of nitronaphthalenes. When mono- and di-nitronaphthalenes are treated with KO2/crown ehter, hydroxylation results at the electron-deficient site caused by the electron withdrawing effect of the substituted nitro group. Kinetic experiments suggest that the hydroxylation proceeds by two different mechanisms dependent on the first one-electron reduction potential of nitronaphthalenes. PMID- 11045463 TI - Rate enhancement with high ratio of the monoalkylated product to the dialkylated product in the alkylation of the lithium enolate of 1-tetralone with reactive alkyl halides. AB - The reactions of the lithium enolate of 1-tetralone with reactive alkyl halides were examined in the absence and in the presence of 3 eq of various ligands for the lithium. It is shown that the rates of the reactions are enhanced greatly in the presence a tetradentate amine (1,1,4,7,10,10-hexamethyltriethylenetetramine), and the ratio of the monoalkylated product to the dialkylated product is increased under shorter reaction times. PMID- 11045465 TI - Insight into acid-mediated asymmetric spirocyclization in the presence of a chiral diol. AB - Asymmetric spirocyclization based on intramolecular conjugate addition using a combination of a Lewis acid and an optically active cyclohexane-1,2-diol has been studied in connection with 1) the effect of substituents on the cyclohexane-1,2 diol and 2) the effect of substituents on the substrate. This reaction was found to be both thermodynamically and kinetically controlled under restricted conditions. PMID- 11045466 TI - Sterically constrained 'roofed' 2-thiazolidinones as excellent chiral auxiliaries. AB - The "roofed" chiral 2-thiazolidinones, which are sterically congested and conformational rigid, and which are prepared by the [4+2] cycloaddition of 2 thiazolone to the cyclic dienes, dimethylanthracene and hexamethylcyclopentadiene, followed by optical resolution with (1S,2R)-2-methoxy 1-apocamphanecarbonic acid (MAC acid) are of considerable promise for use as chiral auxiliaries for the alkylation of enolates. PMID- 11045467 TI - Construction of an enantiomerically pure 6-substituted 3,5-syn-dihydroxyhexanoic acid system by an enantioselective deprotonation strategy: formal synthesis of an antiobesity agent, (-)-tetrahydrolipstatin. AB - Preparation of 6-substituted 4-hydroxytetrahydro-2H-pyran-2-one (12), a key intermediate for the synthesis of (-)-tetrahydrolipstatin, was achieved in an optically pure form by employing an enantioselective deprotonation reaction of the prochiral bicyclic derivative (5) as a key step. PMID- 11045468 TI - A practical synthesis of the ABC ring model of ecteinascidins. AB - A practical synthesis of 1,2,3,4,5,6-hexahydro-1,5-imino-10-hydroxy-9-methoxy 3,8,11-trimethyl-3- benzazocin-4-one (3) as an ABC ring model compound of ecteinascidin 743 and safracins from 3-hydroxy-4-methoxy-5-methylbenzaldehyde (7) is described. The overall yield in 15 steps is 27%. PMID- 11045469 TI - Preparation of benzene, furan, and thiophene analogs of duocarmycin SA employing a newly-devised phenol-forming reaction. AB - Five A-ring analogs of duocarmycin SA 9a-e were synthesized in racemic form modifying our second synthetic route toward duocarmycin SA. The problem encountered at the crucial phenol forming step to secure 17a, b from 16a, b under the conventionally used Kuwajima conditions was overcome by devising a more convenient method: simple heating of 16a-c in benzene in the presence of bis(triphenylphosphine)palladium(II) chloride (10 mol%), cesium carbonate (3 eq), and triphenylphosphine (0.3 eq) gave 17a-c in high yields of 86-91%. The intermediates 17a-e were readily led to the A-ring analogs (+/-)-9a-e almost according to the reported route. PMID- 11045470 TI - Enantioselective synthesis of the key intermediate of the acyl-CoA: cholesterol acyltransferase (ACAT) inhibitor (R-106578) using 2,2'-bis(diphenylphosphino) 1,1'-binaphthyl (BINAP)-Ru(OAc)2 as a catalyst. AB - Acidic segment of an acyl-CoA: cholesterol acyltransferase (ACAT) inhibitor, R 106578 was synthesized by enantioselective hydrogenation of the Z-olefine (9-(Z)) using (R)-2,2'-bis(diphenylphosphino)-1,1'-binaphthyl (BINAP)-Ru(OAc)2 as a catalyst in methanol at 100 degrees C, 5 kgf/cm2 of H2 pressure. The requisite Z olefine was prepared regioselectively via coumarin derivative (5). PMID- 11045471 TI - Total synthesis of polyamine toxin HO-416b and Agel-489 using a 2 nitrobenzenesulfonamide strategy. AB - Total synthesis of spider toxins HO-416b (1) and Agel-489 (2) was accomplished using the 2-nitrobenzenesulfonamide (Ns) group as both a protecting and activating group. In this strategy, the C-N bonds were constructed by alkylation of sulfonamides with alkyl halides or Mitsunobu reaction with the corresponding alcohol. Beginning with monoprotection of the symmetrical diamine, the construction of the backbone from diamine 3 was efficiently accomplished in 7 steps for 14 and 9 steps for 29. Removal of the Ns group while the substrate was attached to a novel solid support enabled the efficient isolation of this highly polar compound. PMID- 11045472 TI - Controlling factors in chiral bisoxazoline-catalyzed asymmetric lithium ester enolate-imine condensation producing a beta-lactam. AB - A catalytic amount of external chiral bisoxazoline ligand 3a bearing an isopropyl group as a stereocontrolling group catalyzed a reaction of a lithium ester enolate 4b, generated from 3-pentyl 2-methylpropionate, with benzaldehyde anisidine-imine 5 to afford corresponding beta-lactam 6 in higher 70% ee than that obtained by the reaction using a stoichiometric amount of the ligand. A bulkier ligand 3d bearing a phenyl group gave 81% and 6% ees in stoichiometric and catalytic reactions, respectively. Examination of the varying factors suggested the involvement of mixed aggregates as a reactive species. A working model is presented for prediction of the sense of asymmetric induction. PMID- 11045473 TI - Preparation of novel synthons, uniquely functionalized tetrahydrofuran and tetrahydropyran derivatives. AB - The dianion of the acetoacetic ester reacts with epibromohydrin derivatives to afford a mixture of (Z)-2-alkoxycarbonylmethylidenetetrahydrofuran derivative and (E)-2-alkoxycarbonylmethylidenetetrahydropyran derivative. The selective formation of the tetrahydrofuran derivative is achieved by the use of LiClO4 as the additive. The preparation of the optically active tetrahydrofuran derivatives and tetrahydropyran derivatives is also examined, and the optical purity and absolute configuration of the products is elucidated. PMID- 11045474 TI - A catalytic asymmetric strecker-type reaction promoted by Lewis acid-Lewis base bifunctional catalyst. AB - A general asymmetric Strecker-type reaction is reported, catalyzed by the Lewis acid-Lewis base bifunctional catalyst 1. The reaction of trimethylsilyl cyanide (TMSCN) with various fluorenyl imines, including n-aldimines and alpha,beta unsaturated imines, proceeds with good to excellent enantioselectivities in the presence of a catalytic amount of phenol as additive (20 mol%) (catalytic system 1). The products were successfully converted to the corresponding amino acid derivatives in high yields without loss of enantiomeric purity. Furthermore, hydrogenation or dihydroxylation of the products from alpha,beta-unsaturated imines afforded saturated or functionalized aminonitriles also without loss of enantiomeric purity. The absolute configuration of the products and a control experiment using catalyst 2 supported the proposed dual activation of the imine and TMSCN by the Lewis acid (Al) and the Lewis base moiety (phosphine oxide) of 1. From the mechanistic studies including kinetic and NMR experiments of the catalytic species, the role of PhOH seems to be a proton source to protonate the anionic nitrogen of the intermediate. Specifically, we have found that TMSCN is more reactive than HCN in this catalytic system, probably due to the activation ability of the phosphine oxide moiety of 1 toward TMSCN. This fact prompted us to develop the novel catalytic system 2, consisting of 1 (9 mol%), TMSCN (20 mol%) and HCN (1.2 mol eq). This new system afforded comparable results with obtained by system 1 (1 (9 mol%)-TMSCN (2 mol eq)-PhOH (20 mol%)). PMID- 11045475 TI - Construction of chiral 1,2-cycloalkanopyrrolidines from L-proline using ring closing metathesis (RCM). AB - An efficient synthetic method for the preparation of optically active pyrroloazocine, pyrroloazepine, quinolizidine, indolizidine using ring closing olefin metathesis (RCM) is described. PMID- 11045476 TI - Fast gas chromatography: packed column solvating gas chromatography versus open tubular column gas chromatography. AB - Packed capillary column solvating gas chromatography (SGC) and open tubular column gas chromatography (GC) were compared with respect to their potentials for fast separations. A recently introduced "universal" peak capacity equation was used to compare the performance of these two methods. The effects of various factors on peak capacity were investigated. Results demonstrate that retention factor and column efficiency are the main factors affecting peak capacity for fast separations. Packed columns produce both high retention factors and high selectivities. While high efficiencies and high peak capacities can be demonstrated by both techniques, open tubular column GC can surpass packed capillary column SGC in both measurements, except for the case of the analysis of simple mixtures in short analysis times, where retention factor and selectivity become important. Practical aspects such as pressure drop and sample capacity are compared for SGC and open tubular column GC. It was found that packed column SGC demonstrates higher sample capacities, but requires much higher column inlet pressures than open tubular column GC. A variety of mobile phases can be used for packed column SGC, which can provide high solvating power for large and polar compounds. PMID- 11045477 TI - Time-resolved cryogenic modulation for targeted multidimensional capillary gas chromatography analysis. AB - Multidimensional gas chromatography (MDGC) is performed in a new manner, described in this paper. The method incorporates two directly coupled columns and employs a longitudinally modulated cryogenic trap located between the columns. No heartcutting process is used, but rather a method better termed selected zone compression pulsing is used. Compared with normal MDGC, where primary column effluent has to be temporarily diverted either to a monitor detector or to the second dimension column, the new procedure in its simplest mode passes all of the first column effluent to the second column. It is simply the times at which the modulation of the trap is performed that determines which target solutes will be selected for enhanced separation. This approach allows almost instantaneous separation of selected zones on the second column, and has the potential to significantly simplify the MDGC method. Since data are presented in a time response format, and do not require transformation as previously described for comprehensive GC when using the longitudinal modulator, quantitation and report generation are essentially the same as in any GC method and data system. Advantages also include significant sensitivity improvement. By using cryofocussing, and benefiting from the zone compression effects along with fast GC conditions on the second dimension, new possibilities for MDGC can be realised. The method is demonstrated by using a mixture of semi-volatile aromatic hydrocarbons. PMID- 11045478 TI - Comparison of comprehensive two-dimensional gas chromatography and gas chromatography--mass spectrometry for the characterization of complex hydrocarbon mixtures. AB - In this paper, we compare the current separation power of comprehensive two dimensional gas chromatography (GCxGC) with the potential separation power of GC mass spectrometry (GC-MS) systems. Using simulated data, we may envisage a GC-MS contour plot, that can be compared with a GCxGC chromatogram. Real examples are used to demonstrate the current potential of the two techniques in the field of hydrocarbon analysis. As a separation technique for complex hydrocarbon mixtures, GCxGC is currently about as powerful as GC-MS is potentially powerful. GC-MS has not reached its potential separation power in this area, because a universal, soft ionization method does not exist. The greatest advantage of GCxGC is, however, its potential for quantitative analysis. Because flame-ionisation detection can be used, quantitative analysis by GCxGC is much more robust, reliable and reproducible. PMID- 11045479 TI - Synthesis of spherical porous silicas in the micron and submicron size range: challenges and opportunities for miniaturized high-resolution chromatographic and electrokinetic separations. AB - Classical silica technology has reached its limit with respect to an ultimate minimum particle size of about 2 microm in diameter. Here, a novel process is presented which allows one to synthesize porous silica beads and control their particle diameter in situ, within the range of 0.2-2.0 microm. As a result, no sizing is required and losses of silica are avoided. Furthermore, the process enables one to control in situ the pore structural parameters and the surface chemistry of the silica beads. Even though surface funtionalized silicas made according to this process can principally be applied in fast HPLC the column pressure drop will be high even for short columns. In addition, the column efficiency, expressed in terms of the theoretical plate height is about H-2d(p) in the best case and limited by the A and C term of the Van Deemter equation. In other words the gain in total plate number when using 1-2 microm silica beads in short columns is minimal as compared to longer columns packed with 5 microm particles. Capillary electrochromatography (CEC) as a hybrid method enables the application of micron size as well as submicron size particles. This consequently enhances column efficiency by a factor of 5-10 when compared to HPLC. The use of short CEC columns packed with submicron size silicas provides the basis for fast and efficient miniaturized systems. The most significant feature of CEC as compared to HPLC is that the former allows one to resolve polar and ionic analytes in a single run. An alternative method for miniaturization is capillary electrophoresis (CE) which generates extremely high efficiencies combined with fast analysis. Its application, however, is limited to ionic substances. PMID- 11045480 TI - Chromatographic silanol activity test procedures: the quest for a universal test. AB - Reversed-phase chromatography is the most used and the most studied method of modern liquid chromatography. There is yet no ideal support available for preparing reversed-phase stationary phases, but the vast majority have historically been and are still prepared on microparticulate silica. The silica surface has a number of properties which make it attractive for derivatization, including easily controlled particle size and porosity and mechanical stability. There are several types of surface silanols which have their own unique properties that affect both chemical derivatization reactions and adsorptive interactions with solutes. The relative distribution of these different types of silanols may affect the characteristics of silica-based stationary phases more than the absolute number of surface silanol groups. The relative importance of each of these different types of silanols has not yet been unambiguously established. Free or isolated silanols, internally hydrogen-bonded vicinal silanols, and geminal silanols all have been implicated as the primary reaction and adsorption sites. There are many different synthetic schemes that have been used to block the remaining silanols, and "deactivated" phases are very popular. Unfortunately, there is still no universally agreed upon method to measure the accessibility or interaction of these silanols with solute molecules. Many tests have been proposed, focusing mainly on chromatographic probe molecules, but different tests run on the same column will often show different interactions. We will briefly review the surface chemistry of silica and focus on the multitude of tests that have been proposed. Our focal point will be silanol activity test; other aspects of column performance will not be included. Where possible, comparisons among the methods will be made. PMID- 11045482 TI - Optimization methods in chromatography and capillary electrophoresis. AB - Many methods have been developed in order to optimize the parameters of interest in either chromatography or capillary electrophoresis. In chemometric approaches experimental measurements are performed in such a way that all factors vary together. An objective function is utilized in which the analyst introduces the desired criteria (selectivity, resolution, time of analysis). Simplex methods and overlapping resolution maps are declining. Factorial designs and central composite designs are more and more popular in electrodriven capillary separations since the number of parameters to master is much larger than in either GC or LC. The use of artificial neural networks is increasing. The advantage of chemometrics tools is that no explicit models are required, conversely the number of experiments to perform may be high and boundaries of the domain are not straightforward to draw and the approach does more than is required. When models are available optimization is easier to perform by regression methods. Computer assisted methods in RPLC are readily available and work well but are still in infancy in CE. Linear solvation energy relationships seem a very valuable tool but estimates of coefficients still require many experiments. PMID- 11045481 TI - Determination of 1-(2-methoxyphenyl)piperazine derivatives of isocyanates at low concentrations by temperature-programmed miniaturized liquid chromatography. AB - A temperature-programmed packed capillary LC method with large-volume injection on-column focusing has been developed for screening and determination of 1-(2 methoxyphenyl)piperazine derivatives of airborne toluene-2,4-diisocyanate, toluene-2,6-diisocyanate, hexamethylenediisocyanate and methylenebisphenyl-4,4 diisocyanate, based on sampling methods described in MDHS 25/3. Injection volumes up to 100 microl were successfully loaded onto the 250x0.32 mm I.D. capillary column packed with 3 microm Hypersil ODS particles. The isocyanate derivatives were loaded at 10 degrees C and eluted by a three-step temperature program starting at 10 degrees C for 10 min, followed by a temperature ramp of 2.5 degrees C min(-1) to 45 degrees C and then 9.9 degrees C min(-1) to 90 degrees C. The mobile phase consisted of acetonitrile-acetate buffer (3% triethylamine, pH 4.5) (45:55, v/v). The isocyanate derivatives were dissolved in acetonitrile acetate buffer (3% triethylamine, pH 4.5) (30:70, v/v) to achieve sufficient focusing. The concentration limit of detection of the individual derivatives utilizing an "U" shaped flow cell with a 8.0 mm light path and an injection volume of 100 microl was 44, 87, 43 and 210 pg ml(-1) for toluene-2,6 diisocyanate, hexamethylenediisocyanate, toluene-2,4-diisocyanate and methylenebisphenyl-4,4-diisocyanate, respectively. Within the investigated concentration range, 10-500 ng ml(-1), the linear calibration curves gave correlation coefficients ranging from 0.994 to 0.998. The repeatability of the method with regard to retention time and peak height ranged from 0.3 to 1.1% and 1.1 to 2.3% (n=9) relative standard deviation, respectively. The average recovery of the method, with regard to toluene-2,4-diisocyanate, was 97.7+/-1.6% (n=9). PMID- 11045483 TI - Reversed-phase separation of achiral isomers by varying temperature and either gradient time or solvent strength. AB - The separation of several isomer pairs of widely varied structure was studied as a function of changes in temperature, gradient time, mobile phase pH, column type ("monomeric" vs. "polymeric" C18-silica) and organic solvent (methanol vs. acetonitrile). General conclusions are drawn which may prove useful in future attempts at the separation of these and other achiral isomers. PMID- 11045484 TI - Practitioner's guide to method development in thin-layer chromatography. AB - The authors provide a personal perspective of method development in thin-layer chromatography for the novice and more experienced chromatographer alike. No attempt has been made at a comprehensive survey of the literature. Instead we provide an overview with insights into a smaller number of approaches that the authors have found useful in their own work and indicate the factors responsible for the variation in retention and their control. The main topics covered are the relationship between sorbent chemistry and retention, the selection of primary solvents for mobile phase optimization and mobile phase optimization using the PRISMA and solvation parameter models. PMID- 11045485 TI - Capillary electrophoresis coupled to biosensor detection. AB - The present review highlights some modern aspects of biosensor revelation, a detection method which has already found a large number of applications in healthcare, food industry and environmental analysis. First, the concept of bio recognition, which is at the heart of biosensor technology, is discussed, with emphasis on host-guest-like recognition mechanisms. This detection device has been successfully coupled, in its first applications, to chromatographic columns, which allow a high resolution of complex mixtures of analytes prior to interaction with the biosensing unit. The properties of the transducing elements, which should generate a signal (e.g., electrochemical, thermal, acoustic, optical) of proper intensity and of relative fast rise, are additionally evaluated and discussed. The review then focuses on potential applications of biosensing units in capillary electrophoresis (CE) devices. CE appears to be an excellent separation methodology to be coupled to biosensor detection, since it is based on miniaturized electrophoretic chambers, fast analysis times, complete automation in sample handling and data treatment and requires extremely small sample volumes. Although only a few applications of CE-based biosensors have been described up to the present, it is anticipated that this hyphenated technique could have a considerable expansion in the coming years. PMID- 11045486 TI - Non-aqueous capillary electrophoresis. AB - The benefits of non-aqueous capillary electrophoresis have been described in a number of recent publications. The wide selection of organic solvents, with their very different physicochemical properties, broadens our scope to manipulate separation selectivity. The lower currents present in non-aqueous solvents allow the use of high electric field strengths and wide bore capillaries, the latter in turn allowing larger sample load. In many cases detection sensitivity can also be enhanced. The potential of non-aqueous capillary electrophoresis is discussed throughout the paper, and the feasibility of capillary electrophoresis under non aqueous media is demonstrated with reference to several applications. PMID- 11045487 TI - Background theory and applications of microemulsion electrokinetic chromatography. AB - Microemulsion electrokinetic chromatography (MEEKC) is an electrodriven separation technique. Separations are achieved using microemulsions which are nanometre-sized oil droplets suspended in aqueous buffer. The surface tension between the oil and water components is reduced by covered the oil droplet with an anionic surfactant such as sodium dodecyl sulphate and a co-surfactant such as a short-chain alcohol. This review summarises the various microemulsion types and compositions that have been used in MEEKC. The effects of key operating variables such as pH and temperature are also described. The application areas of MEEKC are also described in some detail. MEEKC has been applied to a wide range of water soluble and insoluble both charged and neutral compounds. Examples are described which include analysis of derivatised sugars, proteins, pesticides and a wide range of pharmaceuticals. At present there are only a limited number of publications describing the use of MEEKC but it is anticipated that this number will increase rapidly in the near future as more awareness of the separation possibilities that MEEKC presents increases. PMID- 11045488 TI - Reversed migration micellar electrokinetic chromatography with off-line and on line concentration analysis of phenylurea herbicides. AB - Three environmentally important phenylurea herbicides (monuron, isoproturon, diuron) were separated in reversed migration micellar electrokinetic chromatography (RM-MEKC) using 50 mM sodium dodecyl sulfate, 50 mM phosphoric acid, and 15 mM gamma-cyclodextrin. Three on-line concentration techniques are then evaluated to increase the detection sensitivity of the RM-MEKC system. Stacking with reverse migrating micelles (SRMM, water as the sample solvent) provided the best results among the focusing techniques studied. Using a z-shaped detection cell, more than 500-fold increase in peak height is obtained. As a sample preparation and off-line concentration method, solid-phase extraction (SPE) that further improved detection sensitivity was used in the analysis of spiked tap and pond water. For example, 1 parts per billion of each herbicide spiked in tap or pond water was detected by MEKC after SPE and SRMM. PMID- 11045489 TI - Novel microfabricated device for electrokinetically induced pressure flow and electrospray ionization mass spectrometry. AB - A novel microchip device for electrospray ionization has been fabricated and interfaced to a time-of-flight mass spectrometer. Fluid is electrokinetically transported through the chip to a fine fused-silica capillary inserted directly into a channel at the edge of the device. Electrospray is established at the tip of the capillary, which assures a stable, efficient spray. The electric potential necessary for electrospray generation and the voltage drop for electroosmotic pumping are supplied through an electrically permeable glass membrane contacting the fluidic channel holding the capillary. The membrane is fabricated on the microchip using standard photolithographic and wet chemical etching techniques. Performance relative to other microchip electrospray sources has been evaluated and the device tested for potential use as a platform for on-line electrophoretic detection. Sensitivity was found to be approximately three orders of magnitude better than spraying from the flat edge of the chip. The effect of the capillary on electroosmotic flow was examined both experimentally and theoretically. PMID- 11045490 TI - Alternative methods providing enhanced sensitivity and selectivity in capillary electroseparation experiments. AB - The study of alternative and novel techniques for altering selectivity and enhancing sensitivity as well as injection and detection protocols are important in the ongoing development of capillary electroseparation protocols. Some recent research from our laboratory in these fields is presented and discussed in this review. To improve sensitivity an off-line sample enrichment technique utilising solvent evaporation in a levitated drop or an on-line solid-phase extraction protocol was used. The selectivity was tuned by the use of protein gels or molecularly imprinted polymer mediated capillary electrochromatography. Furthermore, a picolitre droplet injection method is described as well as a detection protocol based on laser-induced fluorescence imaging. PMID- 11045491 TI - Electrochromatography. AB - Although Mould and Synge were credited with the first demonstration of the use of electroosmotic flow to drive a liquid in a thin-layer chromatography system, it was the work of Knox and Grant that extensively detailed much of both the theoretical and practical implications of capillary electrochromatography. This review is aimed at giving a very broad outline of the progression of this technique since the late 1980s up to the present time, including some of the current limitations. Examples will be given of the analysis of a wide variety of materials including neutral, acidic, basic and chiral pharmaceutical compounds, along with some of the instrumentation used for this technique. PMID- 11045492 TI - Advances in column technology and instrumentation in capillary electrochromatography. AB - Capillary electrochromatography (CEC) is an emerging technique gaining increased interest. Improvement of instrumentation and column technology will be of prime importance for the further development of this technique and its use in validated methods. In this paper, developments in column technology and instrumentation for CEC are reviewed with emphasis on developments within the last 3 years. Attention is directed to the employment of stationary phases specifically designed for CEC, the use of soft and rigid gels in place of packings, fritless packed capillaries, column dimensions, the optimization of injection and detection parameters, and gradient elution CEC. PMID- 11045493 TI - Capillary electrochromatography on silica columns: factors influencing performance. AB - Much capillary electrochromatography (CEC) work is carried out on bonded silica packings which offer many advantages: the number of such packings which are available; the fact that the chemistry of bonding and the separation process are fairly well understood; and the possibility of the transfer to CEC of existing HPLC methods. Packing methods for the preparation of CEC columns have been investigated. The problems inherent in the use of burned-in frits remains an obstacle, but can be at least partially overcome by minimising the length and by silanisation. The influence of a variety of mobile phase variables on aspects of CEC is in agreement with theory for: ionic strength, organic content (including isoeluotropy), and pH. Temperature can be used as a variable to change column selectivity in CEC. The influence of pH on electroosmotic flow (EOF) by changing the degree of ionisation of residual silanol groups is similar for a wide range of neutral bonded groups, but is much less marked for bonded sulphonic acid groups. The EOF may be reversed for bonded groups containing nitrogen. PMID- 11045494 TI - Migration of ions in capillary electrochromatography. AB - For capillary electrochromatography (CEC) to be a generally used analytical technique the origin of the unusual, and often unwanted, peak shapes, which regularly occur for ionic compounds, must be understood. A mass balance analysis is the most fundamental approach to investigate the origin of non-linear effects during the migration of an eluite. Such an analysis shows that a CEC system composed of ionic compounds has a complex behaviour and that a variety of peak shapes for an eluite ion is expected. In this paper it is shown that the mass balance analysis is rationalised by the introduction of the non-dimensional electrochromatographic migration number omega. This number is defined as the ratio Eu/v0k, where E is the effective electric field strength in the eluite zone, u the mobility of the eluite, v0 the linear velocity of the mobile phase and k the chromatographic capacity factor of the eluite. This work is focussed on the theoretical behaviour of a CEC system for analytical applications, i.e., in the limit of low eluite concentrations. Even under analytical conditions the three-component system studied in this paper shows strong peak broadening when omega has values close to unity. PMID- 11045495 TI - Peak shapes in open tubular ion-exchange capillary electrochromatography of inorganic anions. AB - An experimental study of parameters influencing peak shapes in ion-exchange open tubular (OT) capillary electrochromatography (CEC) was conducted using adsorbed quaternary aminated latex particles as the stationary phase. The combination of separation mechanisms from both capillary electrophoresis and ion-exchange chromatography results in peak broadening in OT-CEC arising from both these techniques. The sources of peak broadening that were considered included the relative electrophoretic mobilities of the eluent co-ion and analyte, and resistance to mass transfer in both the mobile and stationary phases. The parameters investigated were the mobility of the eluent co-ion, column diameter, separation temperature and secondary interactions between the analyte and the stationary phase. The electromigration dispersion was found to influence peak shapes to a minor extent, indicating that chromatographic retention was the dominant source of dispersion. Improving the resistance to mass transfer in the mobile phase by decreasing the capillary diameter improved peak shapes, with symmetrical peaks being obtained in a 25 microm I.D. column. However, an increase in temperature from 25 degrees C to 55 degrees C failed to show any significant improvement. The addition of p-cyanophenol to the mobile phase to suppress secondary interactions with the stationary phase did not result in the expected improvement in efficiency. PMID- 11045496 TI - Multiple hyphenation of liquid chromatography with nuclear magnetic resonance spectroscopy, mass spectrometry and beyond. AB - The advent of sensitive and reliable HPLC-NMR and HPLC-MS systems has revolutionised the identification of compounds eluting from chromatographic systems. More recently systems have been described wherein both NMR and MS are used together to provide an immensely powerful means of characterising compounds in chromatographic eluents. Here the construction and application of combined HPLC-NMR-MS systems to the analysis of mixtures of pharmaceuticals, drug metabolites in biological fluids and natural products in plant extracts is reviewed. In addition preliminary work with alternative systems such as HPLC-UV NMR-FTIR-MS is highlighted and the prospects for such complex systems considered. PMID- 11045497 TI - Electron-capture mass spectrometry: recent advances. AB - Electron-capture (EC) is a sensitive and selective ionization technique for mass spectrometry (MS). In the most familiar form of EC, a susceptible analyte (electrophore) is detected after eluting from a gas chromatography (GC) column, where a low attomole detection limit for standards is routine. High-performance liquid chromatography can facilitate sample cleanup prior to detection by GC-EC MS, but carryover and shifts in retention time for the "invisible" analyte can be difficulties. Solid-phase extraction avoids these difficulties, but the degree of cleanup and recovery can be problems. Alternative electrophoric derivatizing reagents are available to help deal with interferences, and new reagents such as "AMACE1" are emerging. Releasable forms of electrophores can be used as tags for labeling macromolecules, motivated by the desire to multiplex ligand-type assays. The conventional, gas-phase ion source for EC is not well-understood, especially the role of wall reactions. Using an electron monochromator to tune the electron energy adds to the selectivity and information provided by EC-MS. High-resolution and tandem EC-MS measurements are emerging. Electron-capture dissociation is a new technique to sequence small- to medium-sized peptides, having the advantage of providing more extensive sequence information relative to other MS techniques. Particle-beam EC-MS tends to be less sensitive than GC-EC-MS, but not always. Recently it was demonstrated that EC-MS can be accomplished on an ordinary laser desorption time-of-flight mass spectrometer, and also by using atmospheric pressure chemical ionization. Two applications are discussed here in detail: bile acids and oxidized phenylalanine. EC-MS is well-established as a useful technique for trace analysis in special cases, and the scope of its usefulness is broadening (qualitative analysis and detection of more polar and larger molecules), based on advances in both the chemical and instrumental aspects of this technique. PMID- 11045498 TI - Pesticide residue analysis in foodstuffs applying capillary gas chromatography with mass spectrometric detection. State-of-the-art use of modified DFG multimethod S19 and automated data evaluation. AB - This paper focuses on recent developments in the author's laboratory and reports on the "ultimate" analysis scheme which has evolved over the last 20 years in our laboratory. This demonstrates the feasibility of screening analyses for pesticide residue identification, mainly by full scan GC-MS, down to the 0.01 ppm concentration level in plant foodstuffs. It is based on a miniaturized DFG S19 extraction applying acetone for extraction followed by liquid-liquid extraction with ethyl acetate-cyclohexane followed by gel permeation chromatography. The final chromatographic determination is carried out with a battery of three parallel operating gas chromatographic systems using effluent splitting to electron-capture and nitrogen-phosphorus detection, one with a SE-54 the other with a OV-17 capillary column and the third one with a SE-54 capillary column and mass selective detection for identification and quantitation. The method is established for monitoring more than 400 pesticides amenable to gas chromatography. These pesticide residues are identified in screening analyses by means of the dedicated mass spectral library PEST.L containing reference mass spectra and retention times of more than 400 active ingredients and also metabolites applying the macro program AuPest (Automated residue analysis on Pesticides) for automated evaluation which runs with Windows based HP ChemStation software. The two gas chromatographic systems with effluent splitting to electron capture and nitrogen-phosphorus detection are used to check the results obtained with the automated GC-MS screening and also to detect those few pesticides which exhibit better response to electron-capture and nitrogen-phosphorus detection than to mass spectrometry in full scan. PMID- 11045499 TI - Determination of pesticides in vegetables using large-volume injection column liquid chromatography-electrospray tandem mass spectrometry. AB - Direct injection of a large volume (900 microl) of a sample extract onto a liquid chromatographic (LC) column, LC separation and electrospray tandem mass spectrometric detection were used for the quantitative analysis of a wide polarity range of pesticides in carrots and potatoes. Rapid sample preparation involved extraction of a small amount of sample (2 g) with a small volume of organic solvent (3 ml), clean-up over a filter and dilution of the organic extract with the aqueous LC eluent. The extraction efficiency for the selected pesticides was studied using methanol, acetone and acetonitrile as solvents. Evaluation of the performance of the overall method, using extraction with acetonitrile and detection in the selected-reaction-monitoring mode, showed excellent linearity in the range of 2-100 microg/kg with limits of detection of 0.5-2 microg/kg for both types of vegetable. With relative standard deviations of the MS peak area measurements of less than 6.5% (n=8) the repeatability of the method was fully satisfactory. PMID- 11045500 TI - Determination of steroid sex hormones and related synthetic compounds considered as endocrine disrupters in water by liquid chromatography-diode array detection mass spectrometry. AB - In this study, a procedure for the determination of various naturally occurring hormones and of some related synthetic chemicals, commonly used for birth control and treatment of certain hormonal disorders and cancers, in water is described. The procedure includes solid-phase extraction of 0.5 l of water and subsequent analysis of the extract by liquid chromatography with diode array detection and mass spectrometric detection in series (LC-DAD-MS). DAD has been performed at 197, 225, and 242 nm for quantification and confirmatory identification purposes. For MS detection two interfaces--electrospray and atmospheric pressure chemical ionization--in both the positive and the negative ion mode have been tested and the MS parameters influencing the MS signal optimized. DAD and MS have been intercompared for selectivity, sensitivity, precision, and linearity of response. Selected conditions have been applied to the determination of six estrogens (17beta-estradiol, estriol, estrone, ethynylestradiol, mestranol, and diethylstilbestrol) and four progestogens (progesterone, levonorgestrel, norethindrone and ethynodiol diacetate) in several types of water bodies, including sewage influents and effluents, surface water and drinking water. Recoveries greater than 83% and detection limits in the ng/l range have been achieved for most compounds. PMID- 11045501 TI - Efficiency through combining high-performance liquid chromatography and high resolution gas chromatography: progress 1995-1999. AB - Progress during the last 5 years in on-line LC-GC and related techniques is reviewed. In normal-phase LC-GC, the wire interface proved to have advantages over the loop type interface. Further investigations on the solvent evaporation process in an uncoated precolumn under conditions of an early vapour exit revealed that the rules for the transfer by the retention gap techniques must be modified. For reversed-phase LC-GC, approaches with a phase transfer compete with direct evaporation. Eluents were extracted into a bed of Tenax located in a programmed-temperature vaporiser and thermally desorbed. Direct evaporation is possible when a hot vaporising chamber is used and solvent/solute separation occurs in a separate compartment, a coated precolumn possibly in combination with packed beds. As a future strategy, LC-GC transfer techniques should be adjusted to those of large volume injection and involve a single device. It is believed that on-column injection/transfer is the choice. This requires that concurrent evaporation in LC-GC is performed by the on-column interface. PMID- 11045502 TI - Comparisons of soxhlet extraction, pressurized liquid extraction, supercritical fluid extraction and subcritical water extraction for environmental solids: recovery, selectivity and effects on sample matrix. AB - Extractions of a polycyclic aromatic hydrocarbon (PAH)-contaminated soil from a former manufactured gas plant site were performed with a Soxhlet apparatus (18 h), by pressurized liquid extraction (PLE) (50 min at 100 degrees C), supercritical fluid extraction (SFE) (1 h at 150 degrees C with pure CO2), and subcritical water (1 h at 250 degrees C, or 30 min at 300 degrees C). Although minor differences in recoveries for some PAHs resulted from the different methods, quantitative agreement between all of the methods was generally good. However, the extract quality differed greatly. The organic solvent extracts (Soxhlet and PLE) were much darker, while the extracts from subcritical water (collected in toluene) were orange, and the extracts from SFE (collected in CH2Cl2) were light yellow. The organic solvent extracts also yielded more artifact peaks in the gas chromatography (GC)-mass spectrometry and GC-flame ionization detection chromatograms, especially compared to supercritical CO2. Based on elemental analysis (carbon and nitrogen) of the soil residues after each extraction, subcritical water, PLE, and Soxhlet extraction had poor selectivity for PAHs versus bulk soil organic matter (approximately 1/4 to 1/3 of the bulk soil organic matter was extracted along with the PAHs), while SFE with pure CO2 removed only 8% of the bulk organic matrix. Selectivities for different compound classes also vary with extraction method. Extraction of urban air particulate matter with organic solvents yields very high concentrations of n- and branched alkanes (approximately C18 to C30) from diesel exhaust as well as lower levels of PAHs, and no selectivity between the bulk alkanes and PAHs is obtained during organic solvent extraction. Some moderate selectivity with supercritical CO2 can be achieved by first extracting the bulk alkanes at mild conditions, followed by stronger conditions to extract the remaining PAHs, i.e., the least polar organics are the easiest organics to extract with pure CO2. In direct contrast, subcritical water prefers the more polar analytes, i.e., PAHs were efficiently extracted from urban air particulates at 250 degrees C, with little or no extraction of the alkanes. Finally, recent work has demonstrated that many pollutant molecules become "sequestered" as they age for decades in the environment (i.e., more tightly bound to soil particles and less available to organisms or transport). Therefore, it may be more important for an extraction method to only recover pollutant molecules that are environmentally-relevant, rather than the conventional attempts to extract all pollutant molecules regardless of how tightly bound they are to the soil or sediment matrix. Initial work comparing SFE extraction behavior using mild to strong conditions with bioremediation behavior of PAHs shows great promise to develop extraction methodology to measure environmentally-relevant concentrations of pollutants in addition to their total concentrations. PMID- 11045503 TI - Recent and future developments of liquid chromatography in pesticide trace analysis. AB - Until recently, the application of liquid chromatography (LC) in pesticide analysis was usually focused on groups of compounds or single compounds for which no suitable conditions were available for analysis with gas chromatography (GC). However, recent developments in both detection and column material technology show that LC significantly enlarged its scope in this field of analysis. Obviously, the most striking example is the rather abrupt transition of LC coupled to mass spectrometric detection (MS) from an experimental and scientifically fashionable technique to a robust, sensitive and selective detection mode rendering LC-MS being increasingly used in pesticide trace analysis. Other recent major developments originate from the innovation of new LC column packing materials, viz. immuno-affinity sorbents, restricted access medium materials and molecular imprinted polymers improving considerably the screening of polar pesticides by means of reversed-phase LC with UV detection. In this review the merits and perspectives of these important LC developments and their impact to current and future applications in pesticide trace analysis are presented and discussed. PMID- 11045504 TI - Time-weighted average sampling of volatile and semi-volatile airborne organic compounds by the solid-phase microextraction device. AB - The ultimate goal of the chemist is to perform sample preparation, and analysis, if possible at the place where a sample is located rather than moving the sample to laboratory, as is common practice in many cases at the present time. This approach eliminates errors and time associated with sample transport and storage and therefore it would result in more accurate, precise and faster analytical data. In addition to portability, two other important features of ideal field sample preparation technique are elimination of solvent use and integration with a sampling step. A method is developed which addresses these requirements for the determination of time-weighted average concentration of gas phase compounds using a solid-phase microextraction device. Quantification of target analytes in air using this method can be carried out without external calibration. The volatile and semi-volatile organic compounds in air diffuse into the fiber coating which is retracted a known distance into its needle housing during the sampling period. The coatings used are poly(dimethylsiloxane) and poly(dimethylsiloxane) divinylbenzene. The sampling rate at which gas phase analytes load onto the fiber is determined for a wide range of hydrocarbons. There is a good agreement between the theoretical and experimental sampling rates. Sampling time ranges from 1 min to 24 h depending on the coating used and its retraction distance. Effect of the flow-rate on the uptake rate by the fiber is studied. The method is tested in the field and compared with National Institute of Occupational Health and Safety Method 1550. Good agreement between the results is obtained. PMID- 11045505 TI - Influence of fibre coating in headspace solid-phase microextraction-gas chromatographic analysis of aromatic and medicinal plants. AB - Solid-phase microextraction (SPME) is a solvent-free technique, which is well established in headspace analysis since it is sensitive, because of the concentration factor achieved by the fibres, and selective, because of different coating materials which can be used. The performance of eight commercially available SPME fibres was compared to evaluate the recoveries of some characteristic components with different polarities and structures present in the headspace of four aromatic and medicinal plants: rosemary (Rosmarinus officinalis L.), sage (Salvia officinalis L.), thyme (Thymus vulgaris L.) and valerian (Valeriana officinalis L.). The relative concentration capacity of each fibre on the same components of each plant was also determined by comparing their abundance with that obtained by classical static-headspace GC. The partition coefficient, K1, between the headspace gaseous phase and SPME polymeric coating, and the relative concentration factors, of some of the characteristic components of the plant investigated dissolved in dibutyl phtalate, were also determined, under rigorously standardised analysis conditions. The results showed that the most effective fibres were those consisting of two components, i.e., a liquid phase (polydimethylsiloxane) and a porous solid (carboxen or divinylbenzene, or both). PMID- 11045506 TI - Enantioselective stopped-flow multidimensional gas chromatography. Determination of the inversion barrier of 1-chloro-2,2-dimethylaziridine. AB - Enantioselective stopped-flow multidimensional gas chromatography (stopped-flow MDGC) is a fast and simple technique to determine enantiomerization (inversion) barriers in the gas phase in a range of delta G#gas(T)=70-200 kJ mol(-1). After complete gas-chromatographic separation of the enantiomers in the first column, gas phase enantiomerization of the heart-cut fraction of one single enantiomer is performed in the second (reactor) column at increased temperature and afterwards this fraction is separated into the enantiomers in the third column. From the observed de novo enantiomeric peak areas a(j), the enantiomerization time t and the enantiomerization temperature T, the enantiomerization (inversion) barrier delta G#gas(T) is determined and from temperature-dependent experiments, the activation enthalpy delta H#gas and the activation entropy delta S#gas are obtained. Enantiomerization studies on chiral 1-chloro-2,2-dimethylaziridine by stopped-flow MDGC yielded activation parameters of nitrogen inversion in the gas phase, i.e., delta G#gas(353 K)=110.5+/-0.5 kJ mol(-1), delta H#gas=71.0+/-3.8 kJ mol(-1) and delta S#gas=-109+/-11 J mol(-1) K(-1). By the complementary method of dynamic gas chromatography (GC), the apparent enantiomerization (inversion) barrier of 1-chloro-2,2-dimethylaziridine in the gas-liquid biphase system was found delta G#app(353 K)=108 kJ mol(-1). The values obtained by stopped-flow MDGC in the gas phase were used to calculate the activation parameters of nitrogen inversion of 1-chloro-2,2-dimethylaziridine in the liquid phase in the presence of the chiral selector Chirasil-nickel(II), i.e.. deltaG#liq(353 K)=106.0+/-0.4 kJ mol(-1), delta H#liq=68.3+/-1.4 kJ mol(-1) and deltaS#liq=-106+/-3.0 J mol(-1) K(-1). PMID- 11045507 TI - Enantiomer separations by nonaqueous capillary electrophoresis using octakis(2,3 diacetyl-6-sulfato)-gamma-cyclodextrin. AB - The newest member of the single-isomer isomer sulfated cyclodextrin family, octakis(2,3-diacetyl-6-sulfato)-gamma-cyclodextrin (ODAS-gamma-CD) was used for the first time as a resolving agent for the nonaqueous capillary electrophoretic separation of the enantiomers of 26 weak base pharmaceuticals in an acidic methanol background electrolyte. The solubility limit of ODAS-gamma-CD at room temperature proved to be 55 mM in this background electrolyte, which afforded good, fast enantiomer separations for most of the basic drugs tested. For all the bases studied, the effective mobilities and separation selectivities were found to follow the predictions of the charged resolving agent migration model of electrophoretic enantiomer separations. The effective mobilities of the weakly binding weak bases remained cationic throughout the entire 0 to 45 mM ODAS-gamma CD concentration range; separation selectivities increased as the ODAS-gamma-CD concentration was increased. The effective mobilities of the moderately binding weak bases became anionic in the 2.5 to 45 mM ODAS-gamma-CD concentration range; separation selectivities first increased as the effective mobilities approached zero, then decreased again as the ODAS-gamma-CD concentration was increased further. The effective mobilities of the strongly binding weak bases became anionic in the 0 to 2.5 mM ODAS-gamma-CD concentration range; separation selectivities decreased as the ODAS-gamma-CD concentration was increased above 2.5 mM. PMID- 11045508 TI - Tert.-butylcarbamoylquinine as chiral ion-pair agent in non-aqueous enantioselective capillary electrophoresis applying the partial filling technique. AB - The potential of tert.-butylcarbamoylquinine as chiral selector (SO) added to a non-aqueous background electrolyte for the capillary electrophoretic separation of the enantiomers of N-derivatized amino acids (selectands, SAs) is evaluated. Separation is based on different ion-pair formation equilibrium constants of (R) and (S) enantiomers of the negatively charged chiral analytes with the positively charged quinine-derived chiral SO and on mobility differences of free and complexed SAs, so that differences in the overall migration behavior of the SA enantiomers result. To suppress problems associated with the high UV absorption of the chiral SO and thus the high detector background in the 'total filling technique', the 'partial filling technique' has been adopted. Several parameters including filling time and length of SO zone, respectively, SO concentration, type of background electrolyte, have been evaluated. Using such an optimized method, for example, (R) and (S) enantiomers of 2,4-dinitrophenyl (DNP)-protected proline could be separated with alpha=1.08, R(S)=6.60, and N=130,000 theoretical plates within 15 min. Similar alpha values, resolution, and efficiencies were observed for other DNP-protected, as well as for diverse, N-derivatized amino acids like N-benzoyl, N-9-fluorenylmethoxycarbonyl, N-3,5 dinitrobenzyloxycarbonyl amino acids. A repeatability study clearly validated the robustness of the method and revealed its practical applicability. PMID- 11045509 TI - Sharing experiences. PMID- 11045510 TI - Health physics consequences of out-patient treatment of non-Hodgkin's lymphoma with 131I-radiolabeled anti-B1 antibody. AB - The Medical University of South Carolina is currently participating in clinical trials of 131I radiolabeled Anti-B1 antibody for treatment of Non-Hodgkin's lymphoma. Under current South Carolina Department of Health and Environmental Control regulatory guidelines,; these patients are required to be admitted to the hospital and to remain as inpatients until the whole body burden is <30 mCi or the exposure rate measured 1 m from the patient is <5 mR h(-1). We demonstrate that these patients can be released in accordance with the new recommended guidelines of the Nuclear Regulatory Commission for the release of patients containing radioactive materials in compliance with all radioactive material and public dose standards. This benefits these patients by reducing their risk of infection and other hospital insults and by reducing the length of hospitalizations. Further, unnecessary hospital admissions are decreased, and the overall cost of healthcare delivery for these patients is significantly reduced. PMID- 11045511 TI - Managing a sound industrial radiography radiation safety program. AB - This article was developed to provide new radiation safety officers with the basic information needed for ensuring safety, security, and control of industrial radiography sources and to discuss licensing requirements and other information pertaining to the management of radiation safety programs associated with these sources. PMID- 11045512 TI - Supervision of site radiography contractors. AB - Gamma radiography is used extensively under conditions of site radiography for non destructive testing of components where it is not reasonably practicable to check these components in a shielded enclosure. This is one of the few occasions when a high activity radioactive source is used outside a shielded enclosure and administrative controls are the principal means of restricting exposure. A company where site radiography is required should carry out certain checks themselves to ensure that other personnel not involved in the radiography are not exposed to ionizing radiation. Prior discussion with the radiography company on their safety systems and equipment and checks by the client before and during radiography will reduce the likelihood of unintentional exposures and ensure that no radioactive materials are left on the site. PMID- 11045513 TI - Design and development of a radioactive gas monitoring system in the positron emission tomography center. AB - This paper presents experiences in developing a simple, inexpensive radioactive gas monitoring system that proved to be practical and useful at a clinical and research Positron Emission Tomography (PET) Center. The system is described from four key aspects: (1) design of radiation detectors and calibrations; (2) use of an existing PC-based hospital-wide facility management network; (3) placement and oversight of real-time PC monitor stations; and (4) modification of a ventilation system in the PET center. The radiation safety oversight efficacy of a real-time PC-based radioactive gas monitoring system in the academic PET center also is discussed. PMID- 11045514 TI - Cerenkov counting. AB - When auditing research laboratories, health physics personnel are often alarmed to find liquid scintillation vials that only contain filters and no cocktail. They are then surprised when lab personnel state that they don't need cocktail to count their samples. While not frequently used by the health physicist, counting of samples by using Cerenkov radiation is valid and advantageous when correctly used. PMID- 11045515 TI - Anticipating and addressing workplace static magnetic field effects at levels <0.5 mT. AB - Magnetic resonance, once a research tool limited to the basic sciences, has experienced an increase in popularity due to its unique ability to analyze certain living systems in vivo. Expanding applications in the biomedical sciences have resulted in magnetic sources being located in research institutions nationally. Space and resource limitations sometimes necessitate siting magnetic resonance units in proximity to other institutional operations. For magnetic field shielding and personnel protection considerations, the generally accepted 0.5 mT (milliTesla) limit for implanted cardiac devices is commonly used as the conservative basis for decisions. But the effects of magnetic fields on equipment can be easily observed at levels far below 0.5 mT, often resulting in concern and apprehension on the part of personnel in the surrounding areas. Responding to recurrent worker concerns spawned by noticeable effects on equipment at exposure levels <0.5 mT can strain finite radiation safety program resources. To enhance the ability to anticipate possible facility incompatibility issues associated with the installation of magnetic sources, a literature review was conducted to summarize documented equipment effects. Various types of equipment were found to be impacted at levels ranging down to perhaps 0.001 mT. Armed with this information, practicing radiation safety professionals can better anticipate facility incompatibility issues and improve their responses to worker concerns initiated by observed effects on equipment. PMID- 11045516 TI - Safe use of mobile phones in hospitals. AB - This research evaluated possibilities to use different types of mobile telephones in the hospital environment by testing the disturbances in medical equipment caused by radiofrequency fields emitted by the phones. The research was carried out by the Finnish Institute of Occupational Health (FIOH) in cooperation with the Technical Research Centre of Finland (VTT), and focused mainly on equipment situated within the Maria Hospital of the City of Helsinki. The Helsinki Hospital District and tile Medical Engineering Centre of the Helsinki University Central Hospital (HUCH) also participated in this project. PMID- 11045517 TI - Determining and reporting fetal radiation exposure from diagnostic radiation. AB - Pregnant women are sometimes exposed to diagnostic ionizing radiation for any number of legitimate medical reasons. When this occurs, it is important to perform an accurate fetal dose estimate using standard methodologies as quickly as possible and to convey this information to the woman's physician, along with a summary of appropriate published medical guidelines. This will provide the physician and the patient with the information necessary to make an informed decision regarding the potential impact of this radiation exposure on the pregnancy. At the same time, it is important to recognize that not all pregnancies end well, and there is a finite chance that legal action could ensue because of this, even if radiation exposure was negligible. For this reason, it is also important to make sure that standard, accepted protocols are followed in calculating radiation exposure and the health physicist provide advice only in his or her areas of competence. PMID- 11045518 TI - Pantex: safety in nuclear weapons processing. AB - The Pantex Plant, located in the Texas panhandle near Amarillo, is a major Department of Energy (DOE) participant in maintaining the safety of the nation's nuclear weapons resources and protecting the employees, public, and environment. With more than 168,000 person-years of operations involving nuclear materials, explosives, and hazardous chemicals, Pantex has maintained a notable safety record. This article overviews the nuclear weapon activities at Pantex and describes their safety culture. PMID- 11045519 TI - The quality evaluation program for plutonium pits at the U.S. DOE Pantex plant. AB - The United States Department of Energy Pantex Plant quality evaluation program for plutonium pits is an extensive program that includes 1) weigh and leak check system; 2) radiography; and 3) dye penetrant testing. Successful completion of these diagnostics qualifies a pit to remain in the active status stockpile program. The use of lead aprons and a robot when handling the plutonium pits minimizes personnel exposures to ionizing radiation. All personnel exposures to ionizing radiation at Pantex Plant are As Low As Reasonably Achievable. PMID- 11045521 TI - Solar activity. PMID- 11045520 TI - UV-A and -B transmission of laboratory gloves. PMID- 11045522 TI - Overview of radiation environments and human exposures. AB - Human exposures to ionizing radiation have been vastly altered by developing technology in the last century. This has been most obvious in the development of radiation generating devices and the utilization of nuclear energy. But even air travel has had its impact on human exposure. Human exposure increases with advancing aircraft technology as a result of the higher operating altitudes reducing the protective cover provided by Earth's atmosphere from extraterrestrial radiations. This increase in operating altitudes is taken to a limit by human operations in space. Less obvious is the changing character of the radiations at higher altitudes. The associated health risks are less understood with increasing altitude due to the increasing complexity and new field components found in high-altitude and space operations. PMID- 11045523 TI - Biological effects of cosmic radiation: deterministic and stochastic. AB - Our basic understanding of the biological responses to cosmic radiations comes in large part from an international series of ground-based laboratory studies, where accelerators have provided the source of representative charged particle radiations. Most of the experimental studies have been performed using acute exposures to a single radiation type at relatively high doses and dose rates. However, most exposures in flight occur from low doses of mixed radiation fields at low-dose rates. This paper provides a brief overview of existing pertinent clinical and biological radiation data and the limitations associated with data available from specific components of the radiation fields in airflight and space travel. PMID- 11045524 TI - Radiation measurements in low Earth orbit: U.S. and Russian results. AB - The radiation environment in low-Earth orbital flights is complex. It is strongly influenced by altitude, orbital inclination, time within a given solar cycle, flight duration, and shielding configuration. At any specified shielded location, both primary and secondary particles generated by nuclear interactions of primary particles with spacecraft structure are present. In addition, there are atmospheric secondary albedo protons and neutrons. No single detector can adequately measure this complex radiation field, and measurements of very high linear energy transfer target fragmentation products are particularly difficult. Crew radiation exposure have exclusively been measured using passive thermoluminescent detectors (TLDs). The cosmonaut exposures on the Mir station, uncorrected for the TLD inefficiency and neutron contribution, have varied from a low of 2.43 cGy to a high of 8.70 cGy. These correspond to dose rates of 144 microGy d(-1) to 468 microGy d(-1). These are consistent with rates observed by the D2 ion-chamber. Using the rates measured by the D1 chamber, dose rates under 4 cm of water vary from about 60 microGy d-1 to about 350 microGy d(-1). There is variation of about a factor of two between the dose rates at various locations in the same module. There is also a variation of dose rates of about a factor two between various modules. The highest astronaut dose for a Shuttle flight (STS-82) was 3.205 cGy with a dose rate of 3,221 microGy d(-1). Neutron contribution could be 36 +/- 15% of the astronaut charged particle dose equivalent. East-West asymmetry of dose rate is significant for spacecrafts that fly in an fixed altitude, such as the International Space Station. PMID- 11045525 TI - Radiation exposure for human Mars exploration. AB - One major obstacle to human space exploration is the possible limitations imposed by the adverse effects of long-term exposure to the space environment. Even before human space flight began, the potentially brief exposure of astronauts to the very intense random solar energetic particle events was of great concern. A new challenge appears in deep-space exploration from exposure to the low intensity heavy-ion flux of the galactic cosmic rays since the missions are of long duration, and accumulated exposures can be high. Because cancer induction rates increase behind low to moderate thicknesses of aluminum shielding, according to available biological data on mammalian exposures to galactic cosmic ray-like ions, aluminum shield requirements for a Mars mission may be prohibitively expensive in terms of mission launch costs. Alternative materials for vehicle construction are under investigation to provide lightweight habitat structures with enhanced shielding properties. In the present paper, updated estimates for astronaut exposures on a Mars mission are presented and shielding properties of alternative materials are compared with aluminum. PMID- 11045526 TI - Overview of aircraft radiation exposure and recent ER-2 measurements. AB - The intensity of the different particles making up atmospheric cosmic radiation, their energy distribution, and their potential biological effect on aircraft occupants vary with altitude, geomagnetic latitude, and time in the sun's magnetic activity cycle. Dose rates from cosmic radiation at commercial aviation altitudes are such that crews working on present-day jet aircraft are an occupationally exposed group with a relatively high average effective dose. Crews of future high speed commercial aircraft flying at higher altitudes would be even more exposed. Present calculations of such exposures are uncertain because knowledge of important components of the radiation field comes primarily from theoretical predictions. To help reduce these uncertainties for high-altitude flight, the National Aeronautics and Space Administration (NASA) and the Department of Energy (DOE) started the Atmospheric Ionizing Radiation (AIR) project. The measurement part of the AIR project is an international collaboration of 12 laboratories placing 14 instruments on multiple flights of a NASA ER-2 aircraft. This paper describes the basic features of cosmic radiation in the atmosphere as they relate to exposure of aircraft occupants and then describes the AIR ER-2 measurements and presents some preliminary results from a series of flights in June 1997. PMID- 11045527 TI - British Airways measurement of cosmic radiation exposure on Concorde supersonic transport. AB - The galactic cosmic radiation field at aircraft operating altitudes is complex, with a large energy range and the presence of all particle types. The calculation of the complex radiation fields is difficult, as is the measurement. British Airways continues to cooperate with the U.K. National Radiological Protection Board in measuring cosmic radiation doses on supersonic and subsonic aircraft using a range of devices. PMID- 11045528 TI - Potential doses to passengers and crew of supersonic transports. AB - Data from a tissue equivalent proportional counter that was flown at altitudes ranging from 60,000 feet to 70,000 feet were used to estimate radiation quality factors at different latitudes. For high LET radiation, Q values of 11 to 14 were calculated for latitude 18 degrees north to 59 degrees north. Dose equivalent rates ranging from 5.2 microSv hr(-1) to 27 microSv hr(-1) were measured. These dose equivalent rates are about twice that computed using a computer code called CARI-4Q. The dose equivalent received during a flight from Los Angeles to Tokyo was computed using CARI-4Q and the result doubled, based on the TEPC to CARI-4Q ratio. Members of the general public, including frequent flyers, would not exceed dose limits recommended by the ICRP. Air crew would not exceed the limits for occupationally exposed persons. However, pregnant air crew, based on a 2 mSv limit to concepti, would exceed the limit after 150 hours flying time. PMID- 11045529 TI - The NIOSH/FAA Working Women's Health Study: evaluation of the cosmic-radiation exposures of flight attendants. Federal Aviation Administration. AB - Air crew are exposed to elevated levels of cosmic ionizing radiation of galactic and solar origin and are among the more highly exposed occupational groups to ionizing radiation in the United States. Depending on flight route patterns, the annual dose may range from 0.2 to 5 mSv. By comparison, the average annual radiation dose equivalent of occupationally exposed adults in the United States is estimated to be 1.1 mSv. Cosmic-radiation dose depends primarily on altitude and geomagnetic latitude and to a lesser degree on solar activity. Although the International Commission on Radiological Protection has recommended that air crew exposures to natural radiation in-flight be treated as occupational exposures, United States flight crew exposures to natural cosmic radiation are not regulated or typically monitored. There are approximately 148,000 air crew (flight deck crew and flight attendants) in the United States. PMID- 11045530 TI - Factors affecting cosmic-ray doses at aircraft altitudes. AB - Cosmic rays make a significant, but not normally a dominant, contribution to the radiation dose of people all over the world. However, doses rise with altitude and the earth's magnetic field means that latitude also becomes important. Solar activity imposes a further, time dependent, variation. This article gives more details of these factors in the context of radiation protection as applied to air travel. PMID- 11045531 TI - European measurements of aircraft crew exposure to cosmic radiation. AB - For more than 5 y, the European Commission has supported research into scientific and technical aspects of cosmic-ray dosimetry at flight altitudes in civil radiation. This has been in response to legislation to regard exposure of aircraft crew as occupational, following the recommendations of the International Commission on Radiological Protection in Publication 60. The response to increased public interest and concern, and in anticipation of European and national current work, within a total of three multi-national, multi-partner research contracts, is based on a comprehensive approach including measurements with dosimetric and spectrometric instruments during flights, at high-mountain altitudes, and in a high-energy radiation reference field at CERN, as well as cosmic-ray transport calculations. The work involves scientists in the fields of neutron physics, cosmic-ray physics, and general dosimetry. A detailed set of measurements has been obtained by employing a wide range of detectors on several routes, both on subsonic and supersonic aircraft. Many of the measurements were made simultaneously by several instruments allowing the intercomparison of results. This paper presents a brief overview of results obtained. It demonstrates that the knowledge about radiation fields and on exposure data has been substantially consolidated and that the available data provide an adequate basis for dose assessments of aircraft crew, which will be legally required in the European Union after 13 May 2000. PMID- 11045532 TI - Assessment of the cosmic radiation exposure on Canadian-based routes. AB - As a result of the recent recommendations of the ICRP-60 and in anticipation of possible regulation on occupational exposure of commercial aircrew, a two-phase investigation was carried out over a 1-y period to determine the total dose equivalent on representative Canadian-based flight routes. In the first phase of the study, dedicated scientific flights on a Northern round-trip route between Ottawa and Resolute Bay provided the opportunity to characterize the complex mixed-radiation field and to intercompare various instrumentation using both a conventional suite of powered detectors and passive dosimetry. In the second phase, volunteer aircrew carried (passive) neutron bubble detectors during their routine flight duties. From these measurements, the total dose equivalent was derived for a given route with a knowledge of the neutron fraction as determined from the scientific flights and computer code (CARI-3C) calculations. This study has yielded an extensive database of over 3,100 measurements providing the total dose equivalent for 385 different routes. By folding in flight frequency information and the accumulated flight hours, the annual occupational exposures of 20 flight crew have been determined. This study has indicated that most Canadian-based domestic and international aircrew will exceed the proposed annual ICRP-60 public limit of 1 mSv y(-1) but will be well below the occupational limit of 20 mSv y(-1). PMID- 11045533 TI - Epidemiologic studies of pilots and aircrew. AB - During flight, pilots and cabin crew are exposed to increased levels of cosmic radiation which consists primarily of neutrons and gamma rays. Neutron dosimetry is not straightforward, but typical annual effective doses are estimated to range between two and five mSv. Higher dose rates are experienced at the highest altitudes and in the polar regions. Mean doses have been increasing over time as longer flights at higher altitudes have become more frequent. Because there are so few populations exposed to neutrons, studies of airline personnel are of particular interest. However, because the cumulative radiation exposure is so low, statistical power is a major concern. Further, finding an appropriate comparison group is problematic due to selection into these occupations and a number of biases are possible. For example, increased rates of breast cancer among flight attendants have been attributed to reproductive factors such as nulliparity and increased rates of melanoma among pilots have been attributed to excessive sun exposure during leisure time activities. Epidemiologic studies conducted over the last 20 y provide little consistent evidence linking cancer with radiation exposures from air travel. PMID- 11045534 TI - Dose limits for astronauts. AB - Radiation exposures to individuals in space can greatly exceed natural radiation exposure on Earth and possibly normal occupational radiation exposures as well. Consequently, procedures limiting exposures would be necessary. Limitations were proposed by the Radiobiological Advisory Panel of the National Academy of Sciences/National Research Council in 1970. This panel recommended short-term limits to avoid deterministic effects and a single career limit (of 4 Sv) based on a doubling of the cancer risk in men aged 35 to 55. Later, when risk estimates for cancer had increased and were recognized to be age and sex dependent, the NCRP, in Report No. 98 in 1989, recommended a range of career limits based on age and sex from 1 to 4 Sv. NCRP is again in the process of revising recommendations for astronaut exposure, partly because risk estimates have increased further and partly to recognize trends in limiting radiation exposure occupationally on the ground. The result of these considerations is likely to be similar short-term limits for deterministic effects but modified career limits. PMID- 11045535 TI - Radiation exposure during air travel: guidance provided by the Federal Aviation Administration for air carrier crews. AB - Air carrier crews are occupationally exposed to ionizing radiation, principally from galactic cosmic radiation. To promote radiation safety in aviation the Federal Aviation Administration has: issued educational material on the nature of the radiation received during air travel; recommended radiation exposure limits for pregnant and nonpregnant aircrew members; developed computer programs that estimate for a given flight profile the amount of galactic radiation received on a current flight or on one flown at any time back to January 1958; published tables that enable aircrew members to estimate possible health risks associated with their occupational exposure to radiation; and conducted research on effects of radiation during pregnancy. References for this material are given in the article. In addition, graphic and tabular data in the article show how galactic radiation levels and the composition of the galactic radiation has changed between 1958 and 1999. Also given are estimates of effective doses received by air travelers on a wide variety of air carrier flights. PMID- 11045536 TI - Regulatory control of air crew exposure to cosmic radiation: the European approach. AB - The current European Directive on radiation protection requires that exposure of air crew to cosmic radiation should be assessed if it is likely to exceed 1 milliSievert per year. The approach to this problem in the European Union is described and the relative merits of experimental measurement and computer based assessment are discussed. The particular importance of protection against cosmic radiation exposure in the case of female air crew during pregnancy is described. PMID- 11045537 TI - Perspectives of those impacted: flight attendant's perspective. AB - New regulations regarding radiation exposure to flight attendants are compared to regulations regarding airlines' "no smoking" policies. The new regulations will not be accepted as wholeheartedly by the industry because the ill effects of radiation are not as tangible as those of cigarettes, and there is a risk of a loss of wages due to restrictions during pregnancy. Nevertheless, the carrier's unions would like to act responsibly to ensure the safety of all flight attendants. PMID- 11045538 TI - Perspectives of those impacted: airline pilot's perspective. AB - The airline pilot operates within an environment that consists of circadian dysrhythmia, reduced atmospheric pressure, mild hypoxia, low humidity, and exposure to sound, vibration, cosmic-radiation, and magnetic-field exposure. These occupational exposures present physiological challenges to the long term health of the airline pilot. In particular, exposure to cosmic radiation and its carcinogenic potential have recently received considerable attention. Given the complexity of the environment and possible synergistic exposures, there is an immediate requirement for comprehensive research into both cosmic-radiation and magnetic-field exposures in airline pilots. In response, the Airline Pilots Association International in conjunction with the Medical University of South Carolina (Department of Biometry and Epidemiology) has initiated an extensive research program into these occupational exposures. These investigations include ground based calculations, flight-dose estimates, epidemiological survey and exposure assessment, and biological marker analysis. PMID- 11045539 TI - Perspectives of those impacted: an airline's perspective. AB - The implementation into national law of Council Directive 96/29 EURATOM requires cosmic radiation exposure of flight and cabin crew to be considered as occupational and appropriate measures be taken. The implications for British Airways are discussed and compliance measures outlined. PMID- 11045540 TI - Do the risks justify action? AB - The key issues that have been raised at the meeting on Cosmic Radiation Doses to Air Crew and Astronauts are identified. The paper summarizes the radiation environment and radiation risks associated with exposures to both groups. Finally, the summary concludes that the risks do justify action. PMID- 11045541 TI - Comment on article by Clouvas et al. PMID- 11045542 TI - Comment on dose rate conversion factors for photon emitters in the soil. PMID- 11045544 TI - Ireland and IAEA to monitor the Irish Sea. International Atomic Energy Agency. PMID- 11045543 TI - Body potassium decline--is the male population doomed? PMID- 11045545 TI - Too many or too few orthopedists? PMID- 11045546 TI - Radiologic case study. Metastatic renal cell carcinoma. PMID- 11045547 TI - DVT prophylaxis after THA is safe and effective. PMID- 11045548 TI - DVT prophylaxis after THA: more hype than help. PMID- 11045549 TI - A practical approach to dealing with bone loss in revision total knee arthroplasty. PMID- 11045550 TI - High tibial osteotomy without internal fixation for medial unicompartmental osteoarthrosis. AB - Between 1982 and 1993, a total of 35 patients underwent high tibial osteotomy for medial unicompartmental osteoarthrosis. The osteotomy was performed high in the tibia without the use of jigs, and internal fixation devices were avoided. Patients were assessed using the British Association for Surgery of the Knee score, and satisfactory results with minimal complications were obtained. Our results indicate that in a select group of patients, high tibial osteotomy preserves bone stock and is performed near the deformity so that excellent correction is achieved and recurrence of varus deformity is avoided. PMID- 11045551 TI - In vitro analysis of periprosthetic strains following total knee arthroplasty. AB - Clinical case studies have disclosed certain risk factors associated with periprosthetic fracture in elderly patients. How the mechanical strength of the distal femur is changed by total knee arthroplasty (TKA) has not been elucidated. Using elderly cadaveric femora, this study evaluated both periprosthetic strains and associated fracture patterns arising from an in vitro simulation of a fall onto the distal femur. The data showed a significant increase in anterior and posterior mechanical strain following TKA. Neither stemless nor stemmed versions of two cemented Howmedica prostheses (Rutherford, NJ) reduced distal femur strains to baseline values. However, neither produced a disproportionate frequency of periprosthetic fractures. Although not formally evaluated herein, bone geometry/density may contribute more profoundly to the occurrence of periprosthetic fracture than the implants tested. PMID- 11045552 TI - Augmenting local bone with Grafton demineralized bone matrix for posterolateral lumbar spine fusion: avoiding second site autologous bone harvest. AB - Mineralization and integrity of the bone graft mass were evaluated among patients having posterolateral fusion. Grafting consisted of a composite of Grafton and "local" autologous bone (n=56) or iliac crest autograft alone (n=52). Mineralization was rated radiographically at baseline and at 3, 6, 12, and 24 months. Integrity was judged as fused or not fused. Mineralization ratings did not differ significantly between groups at any postoperative interval (P values of .25-1.00). The percentage of patients fused was similar in both groups (60% and 56% for Grafton and controls, respectively; P=.83). Fifteen control patients reported donor site pain. These findings warrant further evaluation of this composite. PMID- 11045553 TI - AO tension-band osteosynthesis of displaced olecranon fractures. AB - This study reviewed 31 patients who underwent AO tension-band osteosynthesis of displaced olecranon fractures. Thirteen fractures were comminuted. Postoperative immobilization was short, and the median hospital stay was 3 days. In 2 patients, the stainless steel wire broke and required replacement. In 13 patients, the osteosynthesis material was removed after healing because of pain at the tip of the elbow; this did not influence the final result. Median time out of work was 12 weeks. There was no significant loss of elbow power. There was satisfactory mobility, function, and absence of pain. There were 29 good and 2 fair clinical results. Anatomic reduction was achieved in 24 elbows. Possible arthrosis was detected at follow-up in 5 elbows but these patients had a good clinical result. AO tension-band osteosynthesis of displaced olecranon fractures yields good clinical medium-term results with few serious complications. PMID- 11045554 TI - Functional outcome following surgical treatment of metastatic tumors involving the femur. AB - Thirty-eight patients treated surgically for metastatic tumors of the femur were reviewed to evaluate the effects of surgical treatment on quality of life. Average age at surgery was 59 years. The most common origin site of metastatic tumors was the lung. The intertrochanteric area was involved in 14 patients, followed by the subtrochanteric area in 11, femoral neck in 7, and shaft in 6 patients. Surgery was based on the principles that tumor excision should be maximal, and to permit early postoperative ambulation, internal fixation should be rigid with cement augmentation. Types of internal fixation were applied according to lesion site to allow early ambulation and included intramedullary nailing (11), compression hip screw (9), and Rowe plates (7). Prosthetic replacement was performed in 9 patients. Average performance scores improved from 3.8 preoperatively to 2 postoperatively, based on the Functional Classification of the New York Heart Association. In 31 patients (82%), the degree of pain relief was more than a lot according to the criteria of Kaiko. Thirty-five patients survived >5 months postoperatively. In patients with a single metastatic lesion, survival was 21 months, and in patients with multiple lesions, survival was 10 months. Surgical stabilization of a pathologic lesion involving the femur resulted in improvement in the quality of life, including pain relief and early ambulation. In selected patients with a single metastatic lesion and a low-grade primary tumor, prolonged postoperative survival may be expected. PMID- 11045555 TI - Monitoring of somatosensory and motor evoked potentials during open reduction and internal fixation of pelvis and acetabular fractures. AB - Monitoring of motor and somatosensory evoked potentials provides instantaneous intraoperative assessment of a patient's neurologic status. Monitoring of the sciatic nerve through motor and somatosensory evoked potentials can be used during open reduction and internal fixation of pelvic and acetabular fractures. A review of 12 pelvic and acetabular fractures treated with open reduction and internal fixation was conducted and assessed with a combination of intraoperative motor and somatosensory evoked potential monitoring. Results revealed intraoperative motor evoked potential monitoring was 100% sensitive and 100% specific in predicting postoperative sciatic nerve deficits, whereas somatosensory evoked potentials were not accurate in predicting postoperative sciatic nerve deficits. Combined monitoring of the sciatic nerve with motor and somatosensory evoked potentials is beneficial at predicting postoperative sciatic nerve deficits during open reduction and internal fixation of pelvic and acetabular fractures. PMID- 11045556 TI - Musculoskeletal injuries in earthquake victims: an update on orthopedic management. PMID- 11045557 TI - Effect of intermittent administration of parathyroid hormone on fracture healing in ovariectomized rats. AB - The effects of intermittent administration of parathyroid hormone on fracture healing in ovariectomized rats were examined to evaluate its potential use as a therapeutic agent for osteoporotic fractures. Three months postovariectomy, bilateral tibial shaft fractures were induced and stabilized by intramedullary nailing with Kirschner wires. Saline, 17beta-estradiol (Sigma Chemical Corp, St Louis, Mo), or recombinant human parathyroid hormone (1-84) (Korean Green-Cross Pharm Corp, Seoul, Korea) was given once a day for 30 consecutive days during fracture healing. Fracture healing was assessed by morphometric and mechanical analysis of fracture callus. Intermittent parathyroid hormone administration increased the morphometric and mechanical parameters in a dose-dependent manner. A bone-resorption inhibiting agent, 17beta-estradiol did not offer advantage in terms of fracture healing in ovariectomized rats. Findings suggest intermittent parathyroid hormone administration may benefit osteoporosis and fracture. PMID- 11045558 TI - Irreducible dislocation of a total hip prosthesis: the risk of gentamicin beads. PMID- 11045559 TI - Giant cell tumor of the middle phalanx. PMID- 11045560 TI - Rapidly progressive proximal migration of bipolar endoprosthesis due to osteolysis induced by polyethylene debris. PMID- 11045561 TI - Peroneal nerve palsy following intermittent sequential pneumatic compression. PMID- 11045562 TI - Simultaneous bilateral tibial tubercle avulsion fracture. PMID- 11045563 TI - Contemporary management of adult cervical odontoid fractures. PMID- 11045564 TI - Expression of interleukin-7 and its receptor in thyroid lymphoma. AB - Patho-epidemiological studies have shown that thyroid lymphomas (TL) develop in thyroids affected by chronic lymphocytic thyroiditis (CLTH). Cytokines produced in CLTH might play a pivotal role for lymphomagenesis, because previous reports indicate an important role of cytokines in lymphomagenesis. We examined the expression of interleukin-7 (IL-7), a pleiotropic cytokine that acts mainly on cells of the hematolymphoid system, and IL-7 receptor (IL-7R) in both TL and CLTH by RT-PCR. IL-7-specific transcripts were detected more frequently in TL than in CLTH lesions (p < 0.01). IL-7 expression was higher in TL than in CLTH. IL-7R and the common gamma chain were expressed in all but one TL and in all CLTH lesions, in similar levels. We established the sensitive in situ hybridization (ISH) method for detection of IL-7. ISH for IL-7 revealed cytoplasmic signals among cells in the germinal center and mantle zone of lymphoid follicles and interfollicular areas in the TL and CLTH lesions. In the lymphomatous areas of TL, similar numbers of scattered large and small lymphoid cells expressing IL-7 were found. The number of IL-7-expressing cells counted in 10 fields in TL (119.4 +/- 10.6 cells) was significantly higher than found in CLTH lesions (43.1 +/- 4.6) (p < 0.001). These findings suggest a pathogenetic role for IL-7 in the development of TL. ISH showed that the germinal center cells, interfollicular cells, and lymphoma cells expressed IL-7R, but mantle zone cells did not. Because lymphoid cells in lymphoid follicles and the interfollicular area formed in CLTH expressed IL-7, TL cells might proliferate via their own IL-7 and IL-7R, and via IL-7 from reactive lymphoid cells. PMID- 11045565 TI - Structural and functional denervation of human detrusor after spinal cord injury. AB - The bladder receives an extensive nerve supply that is predominantly cholinergic, but several putative transmitters are present, some of which are colocalized. Previous studies have shown increased levels of sensory nerves, reduced inhibitory transmitters, and structural and functional changes in the excitatory input in unstable bladder conditions. The present study compared the end-organ nerve supply to the bladder in spinal cord injury (SCI) with uninjured controls. Acetylcholinesterase histochemistry and double-label immunofluorescence were used to investigate neurotransmitter content, with confocal laser scanning microscopy to assess colocalization. Organ bath studies provided functional correlates for the structural changes in the excitatory innervation. Control samples had dense innervation of the detrusor containing a diverse range of transmitters. Hyperreflexic SCI samples showed patchy denervation, and areflexic SCI samples were diffusely denervated. Vasoactive intestinal polypeptide-, neuropeptide Y-, neuronal nitric oxide synthase-, and galanin-immunoreactive nerve fibers were reduced from frequent or moderately frequent to infrequent or very infrequent in SCI. Calcitonin gene-related peptide-immunoreactive fibers were infrequent in controls and SCI samples. Patterns of colocalization were unchanged, but significantly fewer fibers expressed more than one transmitter. The subepithelial plexus was markedly reduced and several of the smaller coarse nerve trunks showed no immunoreactivity to the transmitters assessed. There was no reduction in sensitivity to electrical field stimulation of intrinsic nerves in SCI, but the maximum force generated by each milligram of bladder tissue and the peak force as a proportion of the maximum carbachol contraction were significantly reduced and the responses were protracted. There was no significant functional atropine resistant neuromuscular transmission in controls or SCI. The reported findings have clinical implications in the management of chronic SCI and development of new treatments. PMID- 11045566 TI - Numerical aberrations of chromosomes 1 and 17 correlate with tumor site in human gastric carcinoma of the diffuse and intestinal types. Fluorescence in situ hybridization analysis on gastric biopsies. AB - Recent studies predict that tumor aneuploidy plays a direct role in tumor instability. The relationship between interphase cytogenetics, histology, grade, and tumor site was analyzed in 20 primary gastric carcinomas. Using fluorescence in-situ hybridization, the numerical changes of centromeric sequences of chromosomes 1, 3, 10, and 17 were directly analyzed in gastric biopsies. Polysomic copy numbers of chromosomes 1 and 17 were discovered in 63% (10 of 16) and 59% (10 of 17), respectively, of informative cancer cases. Chromosome 3 and 10 signal number changes were found in only 6% (1 of 16) and 13% (1 of 8), respectively, of informative cancer cases. There was a positive correlation between the appearance of polysomic nuclear target sites of chromosomes 1 and 17 (correlation coefficient r = 0.72; p < 0.005). Copy number changes were not significantly related to histologic subtypes of either the Lauren or WHO classifications. However, incidence of cancers having dual polysomic signal number abnormalities for both chromosomes 1 and 17 was significantly correlated to tumor location at the cardia. The data suggests that (i) human gastric cancer appears in two genomic groups that can be reliably diagnosed by fluorescence in situ hybridization on routine biopsy sections, (ii) numerical aberrations of chromosomes 1, 3, 10, and 17 are largely independent of histologic subtypes, and (iii) polysomic copy number abnormalities of chromosomes 1 and 17 correlate to intragastric tumor site and are highest in cardia cancers, suggesting high tumor instability at this particular location. PMID- 11045567 TI - The imprinted gene Peg3 is not essential for tumor necrosis factor alpha signaling. AB - The imprinted gene Peg3 encodes a zinc-finger protein which has been proposed to be involved in tumor necrosis factor alpha (TNF) signaling via an interaction with TNF receptor-associated factor 2 (TRAF2). Primary embryonic fibroblasts derived from mice with a null mutation in Peg3 showed no abnormalities in TNF induced nuclear translocation of nuclear factor kappaB (NF-kappaB) or phosphorylation of the mitogen-activated protein kinases, extracellular signal regulated kinases 1 and 2, c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK), and p38. In addition, the loss of Peg3 function did not increase the sensitivity of the cells to the cytotoxic action of TNF. These results suggest that Peg3 does not play an essential role in TNF signal transduction. PMID- 11045568 TI - DNA methylation and the mechanisms of CDKN2A inactivation in transitional cell carcinoma of the urinary bladder. AB - Alterations of the CDKN2A locus on chromosome 9p21 encoding the p16INK4A cell cycle regulator and the p14ARF1 p53 activator proteins are frequently found in bladder cancer. Here, we present an analysis of 86 transitional cell carcinomas (TCC) to elucidate the mechanisms responsible for inactivation of this locus. Multiplex quantitative PCR analysis for five microsatellites around the locus showed that 34 tumors (39%) had loss of heterozygosity (LOH) generally encompassing the entire region. Of these, 17 tumors (20%) carried homozygous deletions of at least one CDKN2A exon and of flanking microsatellites, as detected by quantitative PCR. Analysis by restriction enzyme PCR and methylation specific PCR showed that only three specimens, each with LOH across 9p21, had bona fide hypermethylation of the CDKN2A exon 1alpha CpG-island in the remaining allele. Like most other specimens, these three specimens displayed substantial genome-wide hypomethylation of DNA as reflected in the methylation status of LINE L1 sequences. The extent of DNA hypomethylation was significantly more pronounced in TCC with LOH and/or homozygous deletions at 9p21 than in those without (26% and 28%, respectively, on average, versus 11%, p < 0.0015). No association of LOH or homozygous deletions at 9p21 with tumor stage or grade was found. The data indicate that DNA hypermethylation may be rare in TCC and that deletions are the most important mechanism for inactivation of the CDKN2A locus. The predominance of allelic loss may be explained by its correlation with genome-wide DNA hypomethylation, which is thought to favor chromosomal instability and illegitimate recombination. PMID- 11045569 TI - An animal model for anaplastic large cell lymphoma in the immunocompetent syngeneic C57Bl/6 mouse. AB - We report on the analysis of a murine anaplastic lymphoid cell line TS1G6, established recently by interleukin (IL)-9 transfection. TS1G6 revealed a highly characteristic pattern of large anaplastic cells with mononuclear, binuclear, or multinuclear cells resembling Hodgkin (H) or Sternberg-Reed (SR) cells. This cell line is tumorigenous after injection of as few as 10(4) lymphoma cells into nude or immunocompetent C57Bl/6 mice and leads to death from progressive disease of all treated animals within a few weeks. The histological analysis of these tumors revealed a diffuse large cell malignant lymphoma that is morphologically almost identical to human anaplastic large cell lymphoma (ALCL). The lymphoma cells did not show overexpression of the anaplastic lymphoma kinase (ALK) gene, which is found in about 50% of the cases of human ALCL. Thus, this model may be an animal model for an important subset of human ALCL. The cytokine profile, which is of the T helper 2 type, showed strong parallels to the human lymphoma counterpart. Mice suffering from such lymphomas could not be cured with a regimen using high dose cyclophosphamide similar to many ALCL patients. Such an animal model for ALCL has not yet been recognized, but may provide the basis for investigating new antitumor immunotherapies in a fully immunocompetent host. PMID- 11045570 TI - Attenuation of pulmonary neuroendocrine differentiation in mice lacking Clara cell secretory protein. AB - During development and injury, pulmonary neuroendocrine (NE) cells may transiently express Clara cell 10 kD protein (CC10), a major product of the nonciliated progenitor cells for normal and neoplastic airway epithelia suggesting a close relationship between the cells. To assess the role of CC10 during NE differentiation, we studied CC10-deficient mouse lungs by immunohistochemistry and digital imaging. The knockout model revealed a lack of the disrupted gene product in the lung. Because NE cells, which occur as solitary cells or in neuroepithelial bodies (NEBS), comprise less than 1% of airway epithelia, we counted foci positive for each of the three NE markers- synaptophysin, calcitonin gene-related peptide (CGRP), and protein gene product (PGP) 9.5--and developed a method to analyze numerous airways in serial sections. Digitized images of slides were segmented with Photoshop imaging software. The length of airway epithelium and total section areas were then measured using MetaMorph image analysis software. A comparable range of NE foci was observed regardless of CC10 expression patterns with all three markers, suggesting that CC10 is not critical for NE ontogenesis. However, discrimination according to size revealed that wild-type lungs harbored 30% to 40% greater synaptophysin- and CGRP-containing NEBs relative to CC10 deficient lungs. We posit that an attenuation of pulmonary NE differentiation afflicts the CC10-deficient state. Our imaging application greatly facilitates the acquisition and analysis of complex structures such as the lung and promises to be a widely applicable technique for assessments of tissue sections. PMID- 11045571 TI - VCAM-1, but not ICAM-1 or MAdCAM-1, immunoblockade ameliorates DSS-induced colitis in mice. AB - Adhesion molecule immunoneutralization is envisioned as a promising therapy for inflammatory bowel disease, but the relative value of selective blockade of different adhesion molecules has not been established. The aims of this study were to measure expression and functional relevance of endothelial intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and mucosal addressin cell adhesion molecule 1 (MAdCAM-1) in leukocyte recruitment in experimental colitis and to compare the therapeutic effectiveness of their selective blockade. For this purpose, cell adhesion molecule expression was measured by the dual radiolabeled antibody technique in mice with dextran sulfate sodium-induced colitis and controls. Leukocyte-endothelial cell interactions were determined in colonic venules by fluorescence intravital microscopy. Therapeutic effects of chronic treatment with anti-ICAM-1, anti-VCAM-1, or anti-MAdCAM-1 antibodies were also assessed. Whereas colonic endothelial ICAM-1 was constitutively expressed and had a mild up-regulation in colitic animals, constitutive expression of VCAM-1 and MAdCAM-1 was low, but markedly increased after induction of colitis. Leukocyte adhesion was abrogated by immunoneutralization of VCAM-1 or MAdCAM-1 but not by treatment with an anti-ICAM 1 antibody. Chronic administration of anti-VCAM-1 antibody, but not anti-ICAM-1 or anti-MAdCAM-1, resulted in significant attenuation of colitis in terms of disease activity index, colon length, ratio of colon weight to length, and myeloperoxidase activity. In conclusion, VCAM-1 plays a central role in leukocyte recruitment in colitis and blockade of this adhesion molecule has higher therapeutic effect than immunoneutralization of ICAM-1 or MAdCAM-1 in this experimental model. PMID- 11045572 TI - Clonal analysis of micronodules in virus C-induced liver cirrhosis using laser capture microdissection (LCM) and HUMARA assay. AB - Most hepatocellular carcinomas (HCC) arise from malignant transformation of regenerative cirrhotic nodules. Because HCC has a very poor prognosis, detection of these premalignant lesions may improve the management of patients with cirrhosis. In this regard, clonal analysis of liver micronodules should be of particular interest in order to differentiate polyclonal regenerative micronodules from monoclonal neoplastic potentially malignant micronodules. To address this issue, 112 micronodules from 15 cases of explanted liver cirrhosis were carefully microdissected from paraffin-embedded tissue using a laser capture microscopy system. Clonal analysis was performed by analyzing X-chromosome inactivation, as indicated by the methylation status of the human androgen receptor gene (HUMARA). For each microdissected micronodule, a large set of pathological features was evaluated and correlated with their clonal status. Clonal analysis showed that 57 micronodules (51%) were monoclonal and 55 (49%) were polyclonal. Prevalence of monoclonal nodules ranged from 25% to 71% according to cases. In all cases, mono- and polyclonal nodules were randomly distributed in the cirrhotic liver. Although the clonal status was not significantly affected by the presence or absence of macronodules in the adjacent liver, size of monoclonal micronodules was significantly larger than size of polyclonal micronodules (mean size of the monoclonal nodules: 3 + 0.1 mm vs mean size of the polyclonal nodules: 2.5 +/- 0.1 mm, p = 0.007). Among the elementary pathological features evaluated, only the presence of iron overload was correlated with a monoclonal status (p = 0.04). In conclusion, clonal analysis of liver cirrhosis shows that 51% of micronodules are monoclonal lesions, supporting the notion that liver cirrhosis is a multineoplastic lesion. Because monoclonality is a marker of neoplasia, cirrhosis with accumulation of monoclonal nodules may be carefully followed, and monoclonal nodules should be screened for additional markers to assess their biological behavior. PMID- 11045573 TI - Expression of transforming growth factor-beta and tumor necrosis factor-alpha in the plasma and tissues of mice with lupus nephritis. AB - Although elevated levels of transforming growth factor-beta (TGF-beta) and tumor necrosis factor-alpha (TNF-alpha) have been implicated in renal disease, the tissue distribution and cellular localization of the induced cytokines is not well established. In this study, we investigated the expression of these cytokines during the progression of lupus nephritis in MRL lpr/lpr mice. The concentration of both cytokines increased in the plasma of these animals in an age-dependent manner, and there was an age-dependent induction of TGF-beta and TNF-alpha mRNAs in their kidneys. Although the increase in TGF-beta mRNA was specific for the kidney, the increase in TNF-alpha mRNA was widespread and also could be demonstrated in the liver, lung, and heart. In situ hybridization analysis of renal tissues from the lupus-prone mice localized TGF-beta mRNA to the glomerulus, and more specifically, to resident glomerular cells and inflammatory cells infiltrating periglomerular spaces in the nephritic lesions. The signals for TNF-alpha mRNA were detected only in inflammatory cells and were distributed throughout the nephritic kidney. Plasminogen activator inhibitor-1 (PAI-1) is known to be elevated in the glomeruli of MRL lpr/lpr mice, and intraperitoneal administration of either TGF-beta or TNF-alpha into normal mice markedly induced the expression of this potent inhibitor of fibrinolysis in renal glomerular or tubular cells in vivo. These results suggest that the increased renal expression of both cytokines may contribute to the development of lupus nephritis in this model and raise the possibility that PAI-1 may be involved. The fact that TGF-beta is specifically induced in the kidney whereas TNF-alpha increases in a variety of tissues, supports the hypothesis that the renal specificity of this disorder reflects the abnormal expression of TGF-beta. PMID- 11045574 TI - E-cadherin and cadherin-associated cytoplasmic proteins are expressed in murine mast cells. AB - Cadherins, calcium-dependent cell adhesion molecules, play crucial roles, not only in the maintenance of tissue integrity, but also in the regulation of many aspects of cell behavior. We investigated the expression of "classic" E-, N- and P-cadherins in bone marrow-derived cultured mast cells (BMMC) and peritoneal mast cells (PMC) from mice. Flow cytometric analysis and immunocytochemical staining indicated that E-cadherin was expressed on the cell surface of BMMC and also at lower levels on PMC. N-cadherin was also expressed on the surface of BMMC, but not of PMC, whereas P-cadherin expression was seen in neither cell type. Significant expression of E- and N-cadherin mRNA was observed in BMMC by reverse transcriptase-polymerase chain reaction (RT-PCR), but PMC expressed only E cadherin mRNA. Western blotting analysis indicated expression of alpha- and beta catenins and p120-catenin (or p120 cas) in BMMC, whereas PMC showed less intense expression of alpha- and beta-catenins with high levels of p120 expression. Analyses of beta-catenin or E-cadherin immunoprecipitates from BMMC lysate revealed that alpha-catenin, beta-catenin, and E-cadherin were co-precipitated, suggesting that E-cadherin and catenins form a complex in mast cells. Addition of a blocking antibody of homophilic E-cadherin interactions, or a synthetic E cadherin-binding decapeptide containing the histidine-alanine-valine (HAV) sequence in methylcellulose cultures of gut intraepithelial mononuclear cells or BMMC, significantly suppressed the clonal growth of mast cells. Furthermore, the blocking antibody or synthetic decapeptide significantly suppressed BMMC adhesion to E-cadherin-expressing F9 cell monolayers. These results indicated that E cadherin and associated cytoplasmic proteins in mast cells might be involved in the regulation of certain stages of mast cell differentiation and cell-cell interactions. PMID- 11045575 TI - Assembly pathway of desmoglein 3 to desmosomes and its perturbation by pemphigus vulgaris-IgG in cultured keratinocytes, as revealed by time-lapsed labeling immunoelectron microscopy. AB - To determine the assembly pathway of desmoglein 3 (Dsg3) into desmosomes and the subsequent effects of pemphigus vulgaris immunoglobulin G (PV-IgG) on such, we employed a time-lapsed labeling for FITC/Rhodamine (Rod) double-stained immunofluorescence and 5-nm/10-nm gold double-stained immunoelectron microscopy by using PV-IgG, which was confirmed to react specifically Dsg3. Cells from a human squamous cell carcinoma cell line (DJM-1) were first treated briefly with PV-IgG (3 min), then incubated in either anti-human IgG-FITC or 5-nm gold antibody-containing medium (5 min), followed by a 60-minute chase in normal medium without antibodies. The same cells were reincubated with PV-IgG medium for 3 minutes, followed by either anti-human IgG-Rod or 10-nm gold antibodies for 5 minutes. Using this method, FITC and 5-nm gold particles show the fate of Dsg3-PV IgG complexes during the following 60-minute chase. IgG-Rod or 10-nm gold particles, which are bound during the last 5 minutes of the chase, show Dsg3 molecules newly expressed on the cell surface during the 60-minute-chase period. Initially, Dsg3 formed two types of small clusters on the nondesmosomal plasma membrane, ie, either half-desmosome-like clusters with keratin intermediate filament (KIF) attachment or simple clusters without KIF attachment. The PV-IgG binding to Dsg3 caused the internalization of the simple clusters into endosomes, but not the half-desmosome-like clusters. After the 60-minute-chase period, both types of cell surface Dsg3 clusters were labeled with only 10-nm gold, suggesting that new Dsg3 molecules were being delivered to the cell surface. Desmosomes were labeled with both 5-nm gold and 10-nm gold, whereas the half-desmosome-like clusters were labeled with only 10-nm gold, suggesting that the desmosomes themselves were not split. These results suggest that Dsg3 first forms simple clusters, followed by KIF-attachment, and then becomes integrated into desmosomes, and that PV-IgG-induced internalization of the nondesmosomal simple clusters of Dsg3 may represent the primary effects of PV-IgG on keratinocytes. PMID- 11045576 TI - Follicular lymphoma with monocytoid B-cell proliferation: molecular assessment of the clonal relationship between the follicular and monocytoid B-cell components. AB - Although a number of studies have recognized that follicular lymphomas may be accompanied by a prominent proliferation of monocytoid B-cells, the clonal relationship between these components has not been adequately assessed. Using laser capture microdissection, we isolated the follicular and monocytoid B-cell components from four well-characterized cases of follicular lymphoma with prominent monocytoid B-cells. DNA from each component was analyzed using polymerase chain reaction (PCR)-based methods to assess for clonal rearrangements of the immunoglobulin heavy chain gene (IgH) and for the presence of the bcl-2 gene major breakpoint region/joining region (MBR/JH) DNA fusion products by conventional PCR and fluorescence melting curve analysis. Evidence of clonal identity was established in the follicular and monocytoid B-cell components of three cases by demonstration of IgH gene rearrangements of identical size using IgH PCR, by comparison of complementarity determining region III (CDRIII) DNA sequences, or by detection of bcl-2 MBR/JH fusion products of identical size and/or melting temperature. Molecular analysis of the fourth case revealed a monoclonal and MBR/JH-positive follicular component accompanied by a polyclonal and MBR/JH-negative monocytoid B-cell proliferation. We conclude that the follicular and monocytoid B-cell components of this variant of follicular lymphoma are clonally identical in the majority of cases. However, in a minority of these cases, the monocytoid B-cell component is reactive. Larger studies that assess the prognostic significance of follicular lymphoma with monocytoid B-cells will benefit from molecular studies that assess the clonal relationship of both components. PMID- 11045577 TI - Ectrodactyly and glaucoma associated with a 7q21.2-q31.2 interstitial deletion. AB - An infant with ectrodactyly, glaucoma, cleft palate, congenital heart defect and genital anomalies associated with a 7(q21.2q31.2) deletion is presented. Glaucoma and ectrodactyly in association with a 7q deletion has not been previously reported. We recommend that early ophthalmological assessment is required in infants with such deletions. PMID- 11045578 TI - Sibs with developmental delay, hirsutism and nail hypoplasia: a new syndrome. AB - A sister and brother with developmental delay, hirsutism and variable nail hypoplasia are described. The facial features of these sibs are striking. We postulate that this represents a new syndrome, the inheritance of which is unknown. PMID- 11045579 TI - Further delineation of the DOOR syndrome. AB - Two related sibships from an extended family have been observed with the features of the DOOR syndrome. These features included deafness, onychodystrophy, osteodystrophy, microcephaly, and global developmental retardation with progressive blindness. Seizures, which were associated with hypsarrhythmia, were frequent and difficult to control and ultimately were the cause of death in two patients. An MRI brain scan of case 1 showed a number of abnormalities including markedly reduced myelination. The urine organic acid analysis showed a ten-fold increase of 2-oxoglutarate. In one patient the placenta was noted to have multiple fluid filled cysts, which is a feature reported in other metabolic diseases. It is suggested that there may be genetic heterogeneity in the syndrome, and the presence of increased 2-oxoglutarate is associated with a more severe phenotype which is frequently lethal. PMID- 11045580 TI - Clinical variability in cerebro-oculo-nasal syndrome: report on two additional cases. AB - Cerebro-oculo-nasal syndrome is a rare multiple congenital anomaly syndrome with structural anomalies of the central nervous system, anophthalmia, and abnormal nares. In this report two additional cases are presented, one of them without central nervous system and/or ocular anomalies. PMID- 11045581 TI - A syndrome with midface asymmetry, defective modelling of the skeleton, catch-up growth and truncal obesity. AB - We report follow-up from birth up to 16 years of age of a patient with a previously undescribed combination of dysmorphic features. These include: intrauterine growth retardation developing to normal adult stature with truncal obesity, asymmetry of the midface skeleton with severe orthodontic problems, brachydactyly of the hands and feet, wide medial phalanges of the fingers, partial soft tissue syndactyly, simian creases and normal mental development. We consider other differential diagnoses and suggest that the patient represents a hitherto undescribed syndrome. PMID- 11045582 TI - Costello syndrome: a cancer predisposing syndrome? AB - Costello Syndrome is a specific MCA/MR syndrome mainly characterized by dysmorphic facial features, peculiar biphasic growth pattern, motor and mental retardation, ectodermal anomalies involving skin and nails, and age dependent development of nasal and perianal papillomata. Heart malformations and/or hypertrophic cardiomyopathy are frequently observed. We report a 4-year-old girl with Costello syndrome who developed an intrathoracic ganglioneuroblastoma. In previous reports two patients with ectodermal tumours have been described, a ganglioneuroblastoma of the adrenal gland and an epithelioma. This third report suggests that neural crest neoplasia may be a significant risk factor for children with Costello syndrome. PMID- 11045583 TI - Cleft lip/palate, abnormal ears, ectrodactyly, congenital heart defect, and growth retardation: definition of the acro-cardio-facial syndrome. AB - We report on a male patient with a constellation of malformations, including cleft lip/palate, abnormal ears, ectrodactyly, congenital heart defect (CHD), and growth retardation. A similar association has been previously reported twice [Richieri-Costa and Orquizas (1987) Rev Brasil Genet X:787-792; Giannotti et al. (1995) J Med Genet 32:72-74], and autosomal recessive inheritance was proposed. PMID- 11045584 TI - A young female with asymmetric manifestations of larsen syndrome: another example of unilateral somatic cell-line mosaicism. AB - Larsen syndrome is characterized by multiple congenital joint dislocations, typical skeletal defects and facial dysmorphism. In this article, we present a female patient with asymmetric Larsen syndrome. We hypothesise that the asymmetric distribution of clinical features in our patient is likely caused by post-zygotic somatic cell-line mosaicism of a dominant gene mutation. PMID- 11045585 TI - Question mark ears, temporo-mandibular joint malformation and hypotonia: auriculo condylar syndrome or a distinct entity? AB - We report a boy with prominent, peculiarly malformed ears, abnormality of the ramus of the mandible and hypotonia. An isolated peculiar bilateral ear deformity named 'question mark ear' has been delineated in plastic reconstruction surgery reviews [Cosman et al., 1970 Plast Reconstr Surg 46:454-457; Cosman (1984) Plast Reconstr Surg 73:572-576; Takato et al. (1989) Ann Plast Surg 22:69-73; Brodovsky (1997) Plast Reconstr Surg 100:1254-1257; Park (1998) Plast Reconstr Surg 101:1620-1623; Al-Quattan (1998) Plast Reconstr Surg 102:439-441] and a similar deformity of the ear and changes in the temporo-mandibular joint and condyle has been described by Jampol et al. [(1998) Am J Med Genet 75:449-452] and by Guion Almeida et al. [(1999) Am J Med Genet 86:130-133]. The present case may be the third description of this malformation complex with additional clinical features characterized by hypotonia and mild developmental delay, or possibly a new distinct entity. PMID- 11045586 TI - MOMO syndrome: a possible third case. AB - This report describes a 5-year-old girl, mildly mentally retarded, with the following characteristics: macrocephaly; severe obesity; ocular abnormalities (right optic disk coloboma and left choroidal coloboma); short stature; and recurvation of the femur. The case is sporadic with no consanguinity between the parents. The condition was diagnosed tentatively as MOMO syndrome (Macrosomia, Obesity, Macrocephaly, and Ocular Abnormalities) (MIM, 157980), because of the presence of short stature, in contrast with the large stature of the only two previously described cases. It is the third possible example of this rare syndrome to be described in the literature, with some new clinical findings presented. PMID- 11045587 TI - Thoracic-pelvic dysostosis. AB - We report a third patient, a female, with thoraco-pelvic dysostosis. This rare disorder is similar in phenotypic and radiographic appearances to thoraco-laryngo pelvic dysplasia (Barnes syndrome). The only major difference between these two diseases is absence of laryngeal involvement in thoraco-pelvic dysplasia. They may represent two different entities or a contiguous gene syndrome. PMID- 11045588 TI - Varadi-Papp syndrome: report of a case. AB - A case of Varadi-Papp syndrome in a 3-year-old boy whose case has been followed from 8 months of age is described. PMID- 11045589 TI - Klippel-Feil syndrome, thenar hypoplasia, carpal anomalies and situs inversus viscerum. AB - A 27-year-old female is described with Klippel-Feil syndrome, thenar hypoplasia, carpal anomalies and situs inversus viscerum. Other anomalies occurring with Klippel-Feil syndrome are discussed. PMID- 11045590 TI - Caudal regression syndrome and annular pancreas: a rare association. AB - A female infant with caudal regression syndrome and annular pancreas is described. This is the first time this association appears to have been described. PMID- 11045591 TI - Oculocutaneous albinism and reduced bone density in two sibs: a new autosomal recessive syndrome? AB - A sister and brother, with oculocutaneous albinism and reduced bone density are described. Autosomal recessive inheritance is possible. This association has not been previously described. PMID- 11045592 TI - Amniotic band sequence versus the autosomal recessive microcephaly, facial clefting, and preaxial polydactyly syndrome. AB - We report a Brazilian boy, born to consanguineous parents. On the left arm there was a proximal 'ring shaped' constriction, regional aplasia cutis, and a short hand with markedly hypoplastic fingers and nails 2-3. He also had a bilateral cleft lip/palate, preaxial polydactyly involving the distal phalanx of the left index finger, and a supernumerary nipple on the right. The differential diagnosis is discussed. PMID- 11045593 TI - Multiple subcutaneous granular-cell tumours in a patient with Noonan syndrome. AB - A boy with Noonan syndrome and multiple subcutaneous granular-cell tumours is described and the association discussed. PMID- 11045594 TI - Overlap between Baller-Gerold and Rothmund-Thomson syndrome. AB - We report a male patient with craniosysnostosis, bilateral radial and ulnar hypoplasia, absent thumbs, poikiloderma, and short stature. His parents are first cousins. Although this patient was originally diagnosed as having Baller-Gerold syndrome it is more likely that he has Rothmund-Thomson syndrome or a similar disorder. This report confirms the overlap between these two syndromes, and that Baller-Gerold syndrome is essentially a diagnosis of exclusion. PMID- 11045595 TI - Congenital microgastria with Pierre Robin sequence and partial trismus. AB - A female with congenital microgastria, Pierre Robin sequence and partial trismus is described. This is a previously undescribed association and the etiology of the association is discussed. PMID- 11045596 TI - Induction of cytokine genes and IL-1alpha by chemical hypoxia in PC12 cells. AB - The role of cytokine in neuronal injury was examined in rat pheochromocytoma (PC12) cells under chemical hypoxia (i.e. KCN) and glucose deprivation. The mRNA levels of interleukin-1alpha (IL-1alpha), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were measured by reverse transcription-polymerase chain reaction (RT PCR) in PC12 cells exposed to 0.5 mM KCN for various time intervals. Cytokine mRNA levels expressed to peak levels 30 minutes after KCN treatment and declined gradually until 240 min. The IL-1alpha activity reached the highest levels 2 hr after the same KCN treatment. Under parallel conditions, KCN increased cytosolic free calcium concentration ([Ca2+]i) in the absence of glucose. However, IL 1alpha mRNA induction by KCN was not altered under calcium-free conditions in PC12 cells, indicating its induction was Ca2+-independent. However, the phosphatidylcholine (PC)-specific phospholipase C (PLC) inhibitor D609 decreased the KCN-induced IL-1alpha mRNA and protein in PC12 cells suggests that PC-PLC might play a role in cytokine induction during hypoxia. PMID- 11045597 TI - Different inhibition of enalaprilat and imidaprilat on bradykinin metabolizing enzymes. AB - Effects of angiotensin-converting enzyme (ACE) inhibitors, enalaprilat and imidaprilat, on bradykinin (BK) metabolizing enzymes, aminopeptidase P (APP), neutral endopeptidase (NEP) and carboxypeptidase N (CPN), were examined. APP activity in the mouse lung was inhibited by enalaprilat in a concentration dependent manner while imidaprilat did not influence the enzyme activity. The inhibitory effects of these ACE inhibitors on the NEP activity in the mouse lung and the CPN activity in the mouse serum were negligible. These data suggested that the influence of enalaprilat on the APP activity and subsequent BK metabolism are different from those of imidaprilat. PMID- 11045598 TI - Effects of sodium and potassium channel blockers on hyperinflation-induced slowly adapting pulmonary stretch receptor stimulation in the rat. AB - The excitatory responses of slowly adapting pulmonary stretch receptor (SAR) activity to hyperinflation (inflation volume = 3 tidal volumes) for approximately 10 respiratory cycles were examined before and after administration of flecainide, a Na+ channel blocker, and 4-aminoprydine (4-AP), a K+ channel blocker. The experiments were performed in anesthetized, artificially ventilated rats after unilateral vagotomy. During hyperinflation the SARs increased their activity during inflation and decreased their discharge during deflation. The magnitude of increased SAR activity during inflation became more prominent as compared to that of decreased receptor activity during deflation. Flecainide treatment (6 mg/kg) that was sufficient to block veratridine (50 microg/kg) induced SAR stimulation did not significantly alter the excitatory response of SAR activity to hyperinflation. Subsequent administration of 3 mg/kg flecainide (a total dose, 9 mg/kg) resulted in a greater inhibition of hyperinflation induced SAR stimulation. Although administration of 4-AP (2 mg/kg) usually stimulated SAR activity, particularly in the deflation phase, in the control ventilation, 4-AP treatment had no significant effect on hyperinflation-induced SAR stimulation. These results suggest that the excitatory effect of hyperinflation on SAR activity may not be involved in the activation of either flecainide-sensitive Na+ channels or 4-AP-sensitive K+ channels. PMID- 11045599 TI - Adrenocorticotropic hormone controls Concanavalin A activation of rat lymphocytes by modulating IL-2 production. AB - The initiation of DNA synthesis and secretion of Interleukin 2 (IL-2) was measured in isolated rat splenic lymphocytes following activation with Concanavalin A (ConA). The extent of 3H-thymidine incorporation into activated cells was tested when cultured with various concentrations of Adrenocorticotropic hormone (ACTH). A paradoxical dose-response curve resulted when ACTH caused a biphasic response of augmenting and inhibiting 3H-thymidine uptake in lymphocytes depending on the hormone concentration. Low levels of ACTH (0.001-1-nM) augmented 3H-thymidine uptake and high levels (10-1000 nM) reversed the effect. The optimal ACTH concentration was 10 pM ACTH in the presence of 5 ug/ml ConA and there was no ACTH effect on quiescent cells (no ConA). Conditioned media from splenic lymphocytes treated with various concentrations of ConA or ACTH was tested for increased uptake of 3H-thymidine by the IL-2 growth dependent Cytotoxic T Lymphocyte Leukemia (CTLL-2) cells. ConA conditioned medium could sustain the CTLL-2 cells indicating the presence of IL-2. Conditioned medium from splenic lymphocytes treated with both ConA and 100 pM ACTH further increased CTLL-2 cell proliferation indicating an additional increase of IL-2 secretion. The identity of IL-2 was confirmed by using an anti-rat IL-2 antibody to neutralize the growth potential of the conditioned medium. ACTH alone had no effect on the CTLL-2 cell proliferation indicating the effect is due solely to induced IL-2 found in the conditioned medium. IL-2 levels in the conditioned media were quantitated by ELISA assay; splenic lymphocytes produced 4.2 ng/ml to ConA only, 19.2 ng/ml in ConA plus 10 nM ACTH, and no detectable IL-2 at ConA plus 10 uM ACTH. These results demonstrated that ACTH modulates IL-2 secretion from activated lymphocytes, which is both biphasic and concentration dependent. PMID- 11045600 TI - Effects of baicalein and wogonin on drug-metabolizing enzymes in C57BL/6J mice. AB - Effects of baicalein and wogonin, the major flavonoids of Scutellariae radix, on cytochrome P450 (CYP), UDP-glucuronosyl transferase (UGT), and glutathione S transferase (GST) were studied in C57BL/6J mice. One-week treatment of mice with a liquid diet containing 5 mM baicalein resulted in 29%, 14%, 36%, 28%, and 46% decreases of hepatic benzo(a)pyrene hydroxylation (AHH), benzphetamine N demethylation (BDM), N-nitrosodimethylamine N-demethylation (NDM), nifedipine oxidation (NFO), and erythromycin N-demethylation (EMDM) activities, respectively. Treatment with a liquid diet containing 5 mM wogonin resulted in 43%, 22%, 21%, 24%, and 35% decreases of hepatic AHH, BDM, NDM, NFO, and EMDM activities, respectively. However, hepatic 7-methoxyresorufin O-demethylation (MROD) activity was increased and decreased by baicalein- and wogonin-treatments, respectively. Similar modulation was observed with caffeine 3-demethylation (CDM) activity. Immunoblot analysis showed that the levels of hepatic CYP2E1 and CYP3A proteins were decreased by both baicalein- and wogonin-treatments. Hepatic CYP1A2 protein level was increased by baicalein but decreased by wogonin. In extrahepatic tissues, renal AHH activity was decreased by wogonin whereas pulmonary AHH, 7-ethoxyresorufin O-deethylation (EROD), and MROD activities were increased by both flavonoids. Both baicalein and wogonin strongly increased CYP1A protein level in mouse lung. Hepatic and renal UGT activities toward p nitrophenol were suppressed by baicalein- and wogonin-treatments. However, cytosolic GST activity was not affected by flavonoids. These results suggest that ingestion of baicalein or wogonin can modulate drug-metabolizing enzymes and the modulation shows tissue specificity. PMID- 11045601 TI - Stress down regulates milk yield in cows by plasmin induced beta-casein product that blocks K+ channels on the apical membranes. AB - Stress and stress related hormones such as glucocorticoids inhibit lactation in cows. In the present study we propose a novel mechanism connecting stress with plasminogen-plasmin system (PPS) (an enzymatic mechanism in milk, which leads to the breakdown of the major milk protein casein). We show that stress activates the PPS leading to an increase in plasmin activity, and that a distinct plasmin induced beta-casein breakdown product (fraction 1-28) is a potent blocker of potassium channels in mammary epithelia apical membranes. The reduction in milk production due to dehydration stress or glucocorticoid (dexamethsone) was correlated with the activities of plasmin and channel blocking activity in the milk of the tested cows. The notion that the axis Stress-PPS-beta-casein fraction 1-28 is responsible for the reduction in milk yield is supported by the results of experiments showing that injecting solution composed of casein digest enriched with beta-casein fraction 1-28 to the udder lumen leads to a transient reduction in milk production. Furthermore, injecting a pure beta-casein fraction 1-28 to the udder lumen of goat's lead also to a transient reduction in milk production with kinetics that was similar to the kinetics observed in cows. PMID- 11045602 TI - Increased activity of the temporal insula in subjects with bradycardia. AB - Though it has been postulated that cortical brain regions participate in the regulation of heart rate, their involvement is poorly understood. Using PET and [18] FDG (to measure regional brain glucose metabolism, which serves as an index of brain function) we compared the regional brain metabolic activity between healthy subjects with bradycardia (<60 beats per minute) with those with normal heart rates in the 75-100 beats per minute range. Statistical Parametric Mapping (SPM) analyses revealed significant differences between the groups predominantly localized to the temporal insula. This finding was corroborated by a separate analysis that measured the metabolic activity for each subject in preselected regions located in the temporal insula. Subjects with bradycardia had significantly higher metabolic activity in the right (p < 0.0001) and in the left temporal insula (p < 0.015) than those with normal heart rates. Moreover, resting heart rates were negatively correlated with metabolism in the right (r = -0.77, p < 0.0001) and in the left temporal insula (r = -0.44, p < 0.05). These results corroborate the importance of the temporal insula in the regulation of resting heart rate in humans. The temporal insula is interconnected with limbic brain region and autonomic centers and suggests that this may be a mechanism by which emotional responses regulate heart rate. PMID- 11045603 TI - Stress, anxiety and peripheral benzodiazepine receptor mRNA levels in human lymphocytes. AB - Peripheral benzodiazepine receptor (PBR) mRNA levels were measured in lymphocytes obtained from a cohort of university students and clinically diagnosed anxious patients. The average level of PBR mRNA was decreased in anxious patients compared to a control group. This data confirms previously published results, but it also indicates that PBR mRNA levels cannot be used as a sole diagnostic measure of anxiety because the range of the individual PBR mRNA levels of the anxious group overlapped the range of the PBR mRNA levels of the control group. PBR mRNA levels in students following academic examinations were increased in some individuals and decreased in others. In the same cohort of students individual levels of cortisol and prolactin were predominantly increased and decreased respectively. There was no correlation between the individual changes in the hormone levels or PBR mRNA, which suggests that each of these parameters is affected by different environmental and physiological factors. Lymphocyte PBR mRNA measurement is a useful additional methodology for studying human stress responses however, its use in clinical studies would require the elucidation of PBR's physiological role. PMID- 11045604 TI - Near-ultraviolet radiation suppresses melatonin synthesis in the chicken retina: a role of dopamine. AB - Effects of near-ultraviolet radiation (UV-A; 325-390 nm, peak at 365 nm) on melatonin content and activity of serotonin N-acetyltransferase (AA-NAT; a key regulatory enzyme in melatonin biosynthesis) were examined in the retina of chickens. Acute exposure of dark-adapted animals to UV-A light produced a marked decline in melatonin content and AA-NAT activity of the retina. The magnitude of the observed changes was dependent upon duration of the light pulse and age of chickens, with 1-2-week old birds being more sensitive to UV-A action than 6-7 week old ones. The decrease in the nocturnal AA-NAT activity evoked by a 5-min UV A pulse gradually deepened during the first 30 min after the return of chickens to constant darkness, then the enzyme activity began to rise, reaching nearly complete restoration within 2.5 hr. Systemic administration to chickens of alpha methyl-p-tyrosine (an inhibitor of catecholamine synthesis; 0.3 g/kg) blocked the suppressive effect of UV-A light on retinal AA-NAT activity. Haloperidol, sulpiride (blockers of D2-family of dopamine (DA) receptors) and 2-chloro-11-(4 methylpiperazino)dibenz[b,f]oxepin (an antagonist of D4-DA receptors), given intraocularly (1-100 nmol/eye) prevented the UV-A light-evoked decrease in AA-NAT activity in the chicken retina in a dose-dependent manner, while raclopride (300 nmol/eye), an antagonist of D2/D3-DA receptors, was ineffective. In dark-adapted chickens exposure to UV-A light increased the DA content of the retina. It is concluded that UV-A radiation, similar to visible light, potently suppresses melatonin biosynthesis in the retina of chicken, with a D4-dopaminergic signal playing the role of an intermediate in this action. PMID- 11045605 TI - Candida albicans and Saccharomyces cerevisiae cell wall mannans produce fever in rats: role of nitric oxide and cytokines. AB - Mannan components of C. albicans (5 mg/kg, i.p.) and S. cerevisiae (2.5 mg/kg, i.p.) cell walls produced pyrogenic responses which were completely inhibited by indomethacin (5 mg/kg, s.c.) pretreatment in rats. A non-selective NOS inhibitor, L-NAME (10 mg/kg, s.c.), also inhibited the pyrogenic effectiveness of C. albicans mannan, whereas it was ineffective on the fever induced by S. cerevisiae mannan. A selective elevation in the serum TNF-alpha levels was observed at the initial phase of the fever due to S. cerevisiae mannan, whereas there was no significant change on the serum levels of TNF-alpha, IL-1beta and IFN-gamma during the latent period or at the initial phase of the fever induced by C. albicans mannan. Injections of N-linked and/or O-linked oligomannosides of the either mannan did not cause any significant change in the body temperature and serum cytokine levels. These data suggest that the mannan components of C. albicans and S. cerevisiae cell walls produce a prostaglandin-dependent fever in rats. The initial signal for fever seems to be different for each mannan. Data also indicate that integrity of the mannans is necessary for the pyrogenic response. PMID- 11045606 TI - The tripeptide-copper complex glycyl-L-histidyl-L-lysine-Cu2+ stimulates matrix metalloproteinase-2 expression by fibroblast cultures. AB - Glycyl-histidyl-lysine-Cu2+ (GHK-Cu) is a tripeptide-copper complex known to be a potent wound healing agent. We previously showed its ability to stimulate in vitro and in vivo the synthesis of extracellular matrix components. The aim of this study was to determine the effects of GHK-Cu on MMP-2 synthesis by dermal fibroblasts in culture. We showed that GHK-Cu increased MMP-2 levels in conditioned media of cultured fibroblasts. This effect was reproduced by copper ions but not by the tripeptide GHK alone. This stimulation was accompanied by an increase of MMP-2 mRNA level. We also showed that GHK-Cu increased the secretion of the tissue inhibitors of metalloproteinases, TIMP-1 and TIMP-2. Taken together, our results underline that GHK-Cu is not only an activator of connective tissue production but also of the remodeling of the extracellular matrix. It is able to modulate MMP expression by acting directly on wound fibroblasts. PMID- 11045607 TI - Functional characterization of the 5'-flanking and the promoter region of the human UCP3 (hUCP3) gene. AB - Uncoupling protein-3 (UCP3) is considered as an important regulator of energy expenditure and thermogenesis in humans. To get insight into the mechanisms regulating its expression we have cloned and characterized about 5 kb of the 5' flanking region of the human UCP3 (hUCP3) gene. 5'-RACE analysis suggested a single transcription initiation site 187 bp upstream from the translational start site. The promoter region contains both TATA and CAAT boxes as well as consensus motifs for PPRE, TRE, CRE and muscle-specific factors like MyoD and MEF2 sites. Functional characterization of a 3 kb hUCP3 promoter fragment in multiple cell lines using a CAT-ELISA identified a cis-acting negative regulatory element between -2983 and -982 while the region between -982 and -284 showed greatly increased basal promoter activity suggesting the presence of a strong enhancer element. Promoter activity was particularly enhanced in the murine skeletal muscle cell line C2C12 reflecting the tissue-selective expression pattern of UCP3. PMID- 11045608 TI - Collectin structure: a review. AB - Collectins are animal calcium dependent lectins that target the carbohydrate structures on invading pathogens, resulting in the agglutination and enhanced clearance of the microorganism. These proteins form trimers that may assemble into larger oligomers. Each polypeptide chain consists of four regions: a relatively short N-terminal region, a collagen like region, an alpha-helical coiled-coil, and the lectin domain. Only primary structure data are available for the N-terminal region, while the most important features of the collagen-like region can be derived from its homology with collagen. The structures of the alpha-helical coiled-coil and the lectin domain are known from crystallographic studies of mannan binding protein (MBP) and lung surfactant protein D (SP-D). Carbohydrate binding has been structurally characterized in several complexes between MBP and carbohydrate; all indicate that the major interaction between carbohydrate and collectin is the binding of two adjacent carbohydrate hydroxyl group to a collectin calcium ion. In addition, these hydroxyl groups hydrogen bond to some of the calcium amino acid ligands. While each collectin trimer contains three such carbohydrate binding sites, deviation from the overall threefold symmetry has been demonstrated for SP-D, which may influence its binding properties. The protein surface between the three binding sites is positively charged in both MBP and SP-D. PMID- 11045609 TI - Methionine oxidation within the cerebroside-sulfate activator protein (CSAct or Saposin B). AB - The cerebroside-sulfate activator protein (CSAct or Saposin B) is a small water soluble glycoprotein that plays an essential role in the metabolism of certain glycosphingolipids, especially sulfatide. Deficiency of CSAct in humans leads to sulfatide accumulation and neurodegenerative disease. CSAct activity can be measured in vitro by assay of its ability to activate sulfatide-sulfate hydrolysis by arylsulfatase A. CSAct has seven methionine residues and a mass of 8,845 Da when deglycosylated. Mildly oxidized, deglycosylated CSAct (+16 Da), separated from nonoxidized CSAct by reversed-phase high-performance liquid chromatography (RP-HPLC), showed significant modulation of the in vitro activity. Because oxidation partially protected against CNBr cleavage and could largely be reversed by treatment with dithiothreitol, it was concluded that the major modification was conversion of a single methionine to its sulfoxide. High resolution RP-HPLC separated mildly oxidized CSAct into seven or more different components with shorter retention times than nonoxidized CSAct. Mass spectrometry showed these components to have identical mass (+16 Da). The shorter retention times are consistent with increased polarity accompanying oxidation of surface exposed methionyl side chains, in general accordance with the existing molecular model. A mass-spectrometric CNBr mapping protocol allowed identification of five of the seven possible methionine-sulfoxide CSAct oxoforms. The most dramatic suppression of activity occurred upon oxidation of Met61 (26% of control) with other residues in the Q60MMMHMQ66 motif falling in the 30-50% activity range. Under conditions of oxidative stress, accumulation of minimally oxidized CSAct protein in vivo could perturb metabolism of sulfatide and other glycosphingolipids. This, in turn, could contribute to the onset and progression of neurodegenerative disease, especially in situations where the catabolism of these materials is marginal. PMID- 11045610 TI - Systematic mutational analysis of the active-site threonine of HIV-1 proteinase: rethinking the "fireman's grip" hypothesis. AB - Aspartic proteinases share a conserved network of hydrogen bonds (termed "fireman's grip"), which involves the hydroxyl groups of two threonine residues in the active site Asp-Thr-Gly triplets (Thr26 in the case of human immunodeficiency virus type 1 (HIV-1) PR). In the case of retroviral proteinases (PRs), which are active as symmetrical homodimers, these interactions occur at the dimer interface. For a systematic analysis of the "fireman's grip," Thr26 of HIV-1 PR was changed to either Ser, Cys, or Ala. The variant enzymes were tested for cleavage of HIV-1 derived peptide and polyprotein substrates. PR(T26S) and PR(T26C) showed similar or slightly reduced activity compared to wild-type HIV-1 PR, indicating that the sulfhydryl group of cysteine can substitute for the hydroxyl of the conserved threonine in this position. PR(T26A), which lacks the "fireman's grip" interaction, was virtually inactive and was monomeric in solution at conditions where wild-type PR exhibited a monomer-dimer equilibrium. All three mutations had little effect when introduced into only one chain of a linked dimer of HIV-1 PR. In this case, even changing both Thr residues to Ala yielded residual activity suggesting that the "fireman's grip" is not essential for activity but contributes significantly to dimer formation. Taken together, these results indicate that the "fireman's grip" is crucial for stabilization of the retroviral PR dimer and for overall stability of the enzyme. PMID- 11045611 TI - A single disulfide bond restores thermodynamic and proteolytic stability to an extensively mutated protein. AB - The potential for engineering stable proteins with multiple amino acid substitutions was explored. Eleven lysine, five methionine, two tryptophan, one glycine, and three threonine substitutions were simultaneously made in barley chymotrypsin inhibitor-2 (CI-2) to substantially improve the essential amino acid content of the protein. These substitutions were chosen based on the three dimensional structure of CI-2 and an alignment of homologous sequences. The initial engineered protein folded into a wild-type-like structure, but had a free energy of unfolding of only 2.2 kcal/mol, considerably less than the wild-type value of 7.5 kcal/mol. Restoration of the lysine mutation at position 67 to the wild-type arginine increased the free energy of unfolding to 3.1 kcal/mol. Subsequent cysteine substitutions at positions 22 and 82 resulted in disulfide bond formation and a protein with nearly wild-type thermodynamic stability (7.0 kcal/mol). None of the engineered proteins retained inhibitory activity against chymotrypsin or elastase, and all had substantially reduced inhibitory activity against subtilisin. The proteolytic stabilities of the proteins correlated with their thermodynamic stabilities. Reduction of the disulfide bond resulted in substantial loss of both thermodynamic and proteolytic stabilities, confirming that the disulfide bond, and not merely the cysteine substitutions, was responsible for the increased stability. We conclude that it is possible to replace over a third of the residues in CI-2 with minimal disruption of stability and structural integrity. PMID- 11045612 TI - Understanding the sequence determinants of conformational switching using protein design. AB - An important goal of protein design is to understand the forces that stabilize a particular fold in preference to alternative folds. Here, we describe an extension of earlier studies in which we successfully designed a stable, native like helical protein that is 50% identical in sequence to a predominantly beta sheet protein, the B1 domain of Streptococcal IgG-binding protein G. We report the characteristics of a series of variants of our original design that have even higher sequence identity to the B1 domain. Their properties illustrate the extent to which protein stability and conformation can be modulated through careful manipulation of key amino acid residues. Our results have implications for understanding conformational change phenomena of central biological importance and in probing the malleability of the sequence/structure relationship. PMID- 11045613 TI - Immucillin-H binding to purine nucleoside phosphorylase reduces dynamic solvent exchange. AB - The rate and extent of hydrogen/deuterium (H/D) exchange into purine nucleoside phosphorylase (PNP) was monitored by electrospray ionization mass spectrometry (ESI-MS) to probe protein conformational and dynamic changes induced by a substrate analogue, products, and a transition state analogue. The genetic deficiency of PNP in humans is associated with severe T-cell immunodeficiency, while B-cell immunity remains functional. Inhibitors of PNP have been proposed for treatment of T-cell leukemia, to suppress the graft-vs.-host response, or to counter type IV autoimmune diseases without destroying humoral immunity. Calf spleen PNP is a homotrimer of polypeptide chains with 284 amino residues, molecular weight 31,541. Immucillin-H inhibits PNP with a Kd of 23 pM when only one of the three catalytic sites is occupied. Deuterium exchange occurs at 167 slow-exchange sites in 2 h when no catalytic site ligands are present. The substrate analogue and product prevented H/D exchange at 10 of the sites. Immucillin-H protected 32 protons from exchange at full saturation. When one of the three subunits of the homotrimer is filled with immucillin-H, and 27 protons are protected from exchange in all three subunits. Deuterium incorporation in peptides from residues 132-152 decreased in all complexes of PNP. The rate and/or extent of deuterium incorporation in peptides from residues 29-49, 50-70, 81-98, and 112-124 decreased only in the complex with the transition state analogue. The peptide-specific H/D exchange demonstrates that (1) the enzyme is most compact in the complex with immucillin-H, and (2) filling a single catalytic site of the trimer reduces H/D exchange in the same peptides in adjacent subunits. The peptides most highly influenced by the inhibitor surround the catalytic site, providing evidence for reduced protein dynamic motion caused by the transition state analogue. PMID- 11045614 TI - Analysis of the internal motion of free and ligand-bound human lysozyme by use of 15N NMR relaxation measurement: a comparison with those of hen lysozyme. AB - Human lysozyme has a structure similar to that of hen lysozyme and differs in amino acid sequence by 51 out of 129 residues with one insertion at the position between 47 and 48 in hen lysozyme. The backbone dynamics of free or (NAG)3-bound human lysozyme has been determined by measurements of 15N nuclear relaxation. The relaxation data were analyzed using the Lipari-Szabo formalism and were compared with those of hen lysozyme, which was already reported (Mine S et al.. 1999, J Mol Biol 286:1547-1565). In this paper, it was found that the backbone dynamics of free human and hen lysozymes showed very similar behavior except for some residues, indicating that the difference in amino acid sequence did not affect the behavior of entire backbone dynamics, but the folded pattern was the major determinant of the internal motion of lysozymes. On the other hand, it was also found that the number of residues in (NAG)3-bound human and hen lysozymes showed an increase or decrease in the order parameters at or near active sites on the binding of (NAG)3, indicating the increase in picosecond to nanosecond. These results suggested that the immobilization of residues upon binding (NAG)3 resulted in an entropy penalty and that this penalty was compensated by mobilizing other residues. However, compared with the internal motions between both ligand-bound human and hen lysozymes, differences in dynamic behavior between them were found at substrate binding sites, reflecting a subtle difference in the substrate-binding mode or efficiency of activity between them. PMID- 11045615 TI - High resolution refinement of beta-galactosidase in a new crystal form reveals multiple metal-binding sites and provides a structural basis for alpha complementation. AB - The unrefined fold of Escherichia coli beta-galactosidase based on a monoclinic crystal form with four independent tetramers has been reported previously. Here, we describe a new, orthorhombic form with one tetramer per asymmetric unit that has permitted refinement of the structure at 1.7 A resolution. This high resolution analysis has confirmed the original description of the structure and revealed new details. An essential magnesium ion, identified at the active site in the monoclinic crystals, is also seen in the orthorhombic form. Additional putative magnesium binding sites are also seen. Sodium ions are also known to affect catalysis, and five putative binding sites have been identified, one close to the active site. In a crevice on the protein surface, five linked five membered solvent rings form a partial clathrate-like structure. Some other unusual aspects of the structure include seven apparent cis-peptide bonds, four of which are proline, and several internal salt-bridge networks. Deep solvent filled channels and tunnels extend across the surface of the molecule and pass through the center of the tetramer. Because of these departures from a compact globular shape, the molecule is not well characterized by prior empirical relationships between the mass and surface area of proteins. The 50 or so residues at the amino terminus have a largely extended conformation and mostly lie across the surface of the protein. At the same time, however, segment 13-21 contributes to a subunit interface, and residues 29-33 pass through a "tunnel" formed by a domain interface. Taken together, the overall arrangement provides a structural basis for the phenomenon of alpha-complementation. PMID- 11045616 TI - Protein engineering as a strategy to avoid formation of amyloid fibrils. AB - The activation domain of human procarboxypeptidase A2 (ADA2h) aggregates following thermal or chemical denaturation at acidic pH. The aggregated material contains well-defined ordered structures with all the characteristics of the fibrils associated with amyloidotic diseases. Variants of ADA2h containing a series of mutations designed to increase the local stability of each of the two helical regions of the protein have been found to have a substantially reduced propensity to form fibrils. This arises from a reduced tendency of the denatured species to aggregate rather than from a change in the overall stability of the native state. The reduction in aggregation propensity may result from an increase in the stability of local relative to longer range interactions within the polypeptide chain. These findings show that the intrinsic ability of a protein to form amyloid can be altered substantially by protein engineering methods without perturbing significantly its overall stability or activity. This suggests new strategies for combating diseases associated with the formation of aggregated proteins and for the design of novel protein or peptide therapeutics. PMID- 11045617 TI - NMR investigation of the interaction of the inhibitor protein Im9 with its partner DNase. AB - The bacterial toxin colicin E9 is secreted by producing Escherichia coli cells with its 9.5 kDa inhibitor protein Im9 bound tightly to its 14.5 kDa C-terminal DNase domain. Double- and triple-resonance NMR spectra of the 24 kDa complex of uniformly 13C and 15N labeled Im9 bound to the unlabeled DNase domain have provided sufficient constraints for the solution structure of the bound Im9 to be determined. For the final ensemble of 20 structures, pairwise RMSDs for residues 3-84 were 0.76 +/- 0.14 A for the backbone atoms and 1.36 +/- 0.15 A for the heavy atoms. Representative solution structures of the free and bound Im9 are highly similar, with backbone and heavy atom RMSDs of 1.63 and 2.44 A, respectively, for residues 4-83, suggesting that binding does not cause a major conformational change in Im9. The NMR studies have also allowed the DNase contact surface on Im9 to be investigated through changes in backbone chemical shifts and NOEs between the two proteins determined from comparisons of 1H-1H-13C NOESY-HSQC spectra with and without 13C decoupling. The NMR-defined interface agrees well with that determined in a recent X-ray structure analysis with the major difference being that a surface loop of Im9, which is at the interface, has a different conformation in the solution and crystal structures. Tyr54, a key residue on the interface, is shown to exhibit NMR characteristics indicative of slow rotational flipping. A mechanistic description of the influence binding of Im9 has on the dynamic behavior of E9 DNase, which is known to exist in two slowly interchanging conformers in solution, is proposed. PMID- 11045618 TI - Conformation and stability of thiol-modified bovine beta-lactoglobulin. AB - Bovine beta-lactoglobulin A assumes a dimeric native conformation at neutral pH, while the conformation at pH 2 is monomeric but still native. Beta-lactoglobulin A has a free thiol at Cys121, which is buried between the beta-barrel and the C terminal major alpha-helix. This thiol group was specifically reacted with 5,5' dithiobis(2-nitrobenzoic acid) (DTNB) in the presence of 1.0 M Gdn-HCI at pH 7.5, producing a modified beta-lactoglobulin (TNB-bIg) containing a mixed disulfide bond with 5-thio-2-nitrobenzoic acid (TNB). The conformation and stability of TNB bIg were studied by circular dichroism (CD), tryptophan fluorescence, analytical ultracentrifugation, and one-dimensional 1H-NMR. The CD spectra of TNB-bIg indicated disordering of the native secondary structure at pH 7.5, whereas a slight increase in the alpha-helical content was observed at pH 2.0. The tryptophan fluorescence of TNB-bIg was significantly quenched compared with that of the intact protein, probably by the energy transfer to TNB. Sedimentation equilibrium analysis indicated that, at neutral pH, TNB-bIg is monomeric while the intact protein is dimeric. In contrast, at pH 2.0, both the intact beta lactoglobulin and TNB-bIg were monomeric. The unfolding transition of TNB-bIg induced by Gdn-HCl was cooperative in both pH regions, although the degree of cooperativity was less than that of the intact protein. The 1H-NMR spectrum for TNB-bIg at pH 3.0 was native-like, whereas the spectrum at pH 7.5 was similar to that of the unfolded proteins. These results suggest that modification of the buried thiol group destabilizes the rigid hydrophobic core and the dimer interface, producing a monomeric state that is native-like at pH 2.0 but is molten globule-like at pH 7.5. Upon reducing the mixed disulfide of TNB-bIg with dithiothreitol, the intact beta-lactoglobulin was regenerated. TNB-bIg will become a useful model to analyze the conformation and stability of the intermediate of protein folding. PMID- 11045619 TI - Tryptophanyl fluorescence lifetime distribution of hyperthermophilic beta glycosidase from molecular dynamics simulation: a comparison with the experimental data. AB - A molecular dynamics simulation approach has been utilized to understand the unusual fluorescence emission decay observed for beta-glycosidase from the hyperthermophilic bacterium Solfolobus sulfotaricus (Sbeta gly), a tetrameric enzyme containing 17 tryptophanyl residues for each subunit. The tryptophanyl emission decay of Sbeta gly results from a bimodal distribution of fluorescence lifetimes with a short-lived component centered at 2.5 ns and a long-lived one at 7.4 ns (Bismuto E, Nucci R, Rossi M, Irace G, 1999, Proteins 27:71-79). From the examination of the trajectories of the side chains capable of causing intramolecular quenching for each tryptophan microenvironment and using a modified Stern-Volmer model for the emission quenching processes, we calculated the fluorescence lifetime for each tryptophanyl residue of Sbeta gly at two different temperatures, i.e., 300 and 365 K. The highest temperature was chosen because in this condition Sbeta gly evidences a maximum in its catalytic activity and is stable for a very long time. The calculated lifetime distributions overlap those experimentally determined. Moreover, the majority of trytptophanyl residues having longer lifetimes correspond to those originally identified by inspection of the crystallographic structure. The tryptophanyl lifetimes appear to be a complex function of several variables, such as microenvironment viscosity, solvent accessibility, the chemical structure of quencher side chains, and side chain dynamics. The lifetime calculation by MD simulation can be used to validate a predicted structure by comparing the theoretical data with the experimental fluorescence decay results. PMID- 11045620 TI - Structure of a (Cys3His) zinc ribbon, a ubiquitous motif in archaeal and eucaryal transcription. AB - Transcription factor IIB (TFIIB) is an essential component in the formation of the transcription initiation complex in eucaryal and archaeal transcription. TFIIB interacts with a promoter complex containing the TATA-binding protein (TBP) to facilitate interaction with RNA polymerase II (RNA pol II) and the associated transcription factor IIF (TFIIF). TFIIB contains a zinc-binding motif near the N terminus that is directly involved in the interaction with RNA pol II/TFIIF and plays a crucial role in selecting the transcription initiation site. The solution structure of the N-terminal residues 2-59 of human TFIIB was determined by multidimensional NMR spectroscopy. The structure consists of a nearly tetrahedral Zn(Cys)3(His)1 site confined by type I and "rubredoxin" turns, three antiparallel beta-strands, and disordered loops. The structure is similar to the reported zinc ribbon motifs in several transcription-related proteins from archaea and eucarya, including Pyrococcus furiosus transcription factor B (PfTFB), human and yeast transcription factor IIS (TFIIS), and Thermococcus celer RNA polymerase II subunit M (TcRPOM). The zinc-ribbon structure of TFIIB, in conjunction with the biochemical analyses, suggests that residues on the beta-sheet are involved in the interaction with RNA pol II/TFIIF, while the zinc-binding site may increase the stability of the beta-sheet. PMID- 11045621 TI - Modeling of loops in protein structures. AB - Comparative protein structure prediction is limited mostly by the errors in alignment and loop modeling. We describe here a new automated modeling technique that significantly improves the accuracy of loop predictions in protein structures. The positions of all nonhydrogen atoms of the loop are optimized in a fixed environment with respect to a pseudo energy function. The energy is a sum of many spatial restraints that include the bond length, bond angle, and improper dihedral angle terms from the CHARMM-22 force field, statistical preferences for the main-chain and side-chain dihedral angles, and statistical preferences for nonbonded atomic contacts that depend on the two atom types, their distance through space, and separation in sequence. The energy function is optimized with the method of conjugate gradients combined with molecular dynamics and simulated annealing. Typically, the predicted loop conformation corresponds to the lowest energy conformation among 500 independent optimizations. Predictions were made for 40 loops of known structure at each length from 1 to 14 residues. The accuracy of loop predictions is evaluated as a function of thoroughness of conformational sampling, loop length, and structural properties of native loops. When accuracy is measured by local superposition of the model on the native loop, 100, 90, and 30% of 4-, 8-, and 12-residue loop predictions, respectively, had <2 A RMSD error for the mainchain N, C(alpha), C, and O atoms; the average accuracies were 0.59 +/- 0.05, 1.16 +/- 0.10, and 2.61 +/- 0.16 A, respectively. To simulate real comparative modeling problems, the method was also evaluated by predicting loops of known structure in only approximately correct environments with errors typical of comparative modeling without misalignment. When the RMSD distortion of the main-chain stem atoms is 2.5 A, the average loop prediction error increased by 180, 25, and 3% for 4-, 8-, and 12-residue loops, respectively. The accuracy of the lowest energy prediction for a given loop can be estimated from the structural variability among a number of low energy predictions. The relative value of the present method is gauged by (1) comparing it with one of the most successful previously described methods, and (2) describing its accuracy in recent blind predictions of protein structure. Finally, it is shown that the average accuracy of prediction is limited primarily by the accuracy of the energy function rather than by the extent of conformational sampling. PMID- 11045622 TI - The oxidation produced by hydrogen peroxide on Ca-ATP-G-actin. AB - We report here that in vitro exposure of monomeric actin to hydrogen peroxide leads to a conversion of 6 of the 16 methionine residues to methionine sulfoxide residues. Although the initial effect of H2O2 on actin is the oxidation of Cys374, we have found that Met44, Met47, Met176, Met190, Met269, and Met355 are the other sites of the oxidative modification. Met44 and Met47 are the methionyl sites first oxidized. The methionine residues that are oxidized are not simply related to their accessibility to the external medium and are found in all four subdomains of actin. The conformations of subdomain 1, a region critical for the functional binding of different actin-binding proteins, and subdomain 2, which plays important roles in the polymerization process and stabilization of the actin filament, are changed upon oxidation. The conformational changes are deduced from the increased exposure of hydrophobic residues, which correlates with methionine sulfoxide formation, from the perturbations in tryptophan fluorescence, and from the decreased susceptibility to limited proteolysis of oxidized actin. PMID- 11045623 TI - Crystal structure of the catalytic domain of human bile salt activated lipase. AB - Bile-salt activated lipase (BAL) is a pancreatic enzyme that digests a variety of lipids in the small intestine. A distinct property of BAL is its dependency on bile salts in hydrolyzing substrates of long acyl chains or bulky alcoholic motifs. A crystal structure of the catalytic domain of human BAL (residues 1-538) with two surface mutations (N186D and A298D), which were introduced in attempting to facilitate crystallization, has been determined at 2.3 A resolution. The crystal form belongs to space group P2(1)2(1)2(1) with one monomer per asymmetric unit, and the protein shows an alpha/beta hydrolase fold. In the absence of bound bile salt molecules, the protein possesses a preformed catalytic triad and a functional oxyanion hole. Several surface loops around the active site are mobile, including two loops potentially involved in substrate binding (residues 115-125 and 270-285). PMID- 11045624 TI - Interaction of mammalian mitochondrial elongation factor EF-Tu with guanine nucleotides. AB - Elongation factor Tu (EF-Tu) promotes the binding of aminoacyl-tRNA (aa-tRNA) to the acceptor site of the ribosome. During the elongation cycle, EF-Tu interacts with guanine nucleotides, aa-tRNA and its nucleotide exchange factor (EF-Ts). Quantitative determination of the equilibrium dissociation constants that govern the interactions of mammalian mitochondrial EF-Tu (EF-Tu(mt)) with guanine nucleotides was the focus of the work reported here. Equilibrium dialysis with [3H]GDP was used to measure the equilibrium dissociation constant of the EF Tu(mt) x GDP complex (K(GDP) = 1.0 +/- 0.1 microM). Competition of GTP with a fluorescent derivative of GDP (mantGDP) for binding to EF-Tu(mt) was used to measure the dissociation constant of the EF-Tu(mt) x GTP complex (K(GTP) = 18 +/- 9 microM). The analysis of these data required information on the dissociation constant of the EF-Tu(mt) x mantGDP complex (K(mGDP) = 2.0 +/- 0.5 microM), which was measured by equilibrium dialysis. Both K(GDP) and K(GTP) for EF-Tu(mt) are quite different (about two orders of magnitude higher) than the dissociation constants of the corresponding complexes formed by Escherichia coli EF-Tu. The forward and reverse rate constants for the association and dissociation of the EF Tu(mt) x GDP complex were determined using the change in the fluorescence of mantGDP upon interaction with EF-Tu(mt). These values are in agreement with a simple equilibrium binding interaction between EF-Tu(mt) and GDP. The results obtained are discussed in terms of the recently described crystal structure of the EF-Tu(mt) x GDP complex. PMID- 11045625 TI - Thermodynamic dissection of the binding energetics of KNI-272, a potent HIV-1 protease inhibitor. AB - KNI-272 is a powerful HIV-1 protease inhibitor with a reported inhibition constant in the picomolar range. In this paper, a complete experimental dissection of the thermodynamic forces that define the binding affinity of this inhibitor to the wild-type and drug-resistant mutant V82F/184V is presented. Unlike other protease inhibitors, KNI-272 binds to the protease with a favorable binding enthalpy. The origin of the favorable binding enthalpy has been traced to the coupling of the binding reaction to the burial of six water molecules. These bound water molecules, previously identified by NMR studies, optimize the atomic packing at the inhibitor/protein interface enhancing van der Waals and other favorable interactions. These interactions offset the unfavorable enthalpy usually associated with the binding of hydrophobic molecules. The association constant to the drug resistant mutant is 100-500 times weaker. The decrease in binding affinity corresponds to an increase in the Gibbs energy of binding of 3 3.5 kcal/mol, which originates from less favorable enthalpy (1.7 kcal/mol more positive) and entropy changes. Calorimetric binding experiments performed as a function of pH and utilizing buffers with different ionization enthalpies have permitted the dissection of proton linkage effects. According to these experiments, the binding of the inhibitor is linked to the protonation/deprotonation of two groups. In the uncomplexed form these groups have pKs of 6.0 and 4.8, and become 6.6 and 2.9 in the complex. These groups have been identified as one of the aspartates in the catalytic aspartyl dyad in the protease and the isoquinoline nitrogen in the inhibitor molecule. The binding affinity is maximal between pH 5 and pH 6. At those pH values the affinity is close to 6 x 10(10) M(-1) (Kd = 16 pM). Global analysis of the data yield a buffer- and pH-independent binding enthalpy of -6.3 kcal/mol. Under conditions in which the exchange of protons is zero, the Gibbs energy of binding is -14.7 kcal/mol from which a binding entropy of 28 cal/K mol is obtained. Thus, the binding of KNI-272 is both enthalpically and entropically favorable. The structure-based thermodynamic analysis indicates that the allophenylnorstatine nucleus of KNI-272 provides an important scaffold for the design of inhibitors that are less susceptible to resistant mutations. PMID- 11045626 TI - Polypeptide stimulators of the Ms-Lon protease. AB - Both the peptidase activity against small fluorescent peptide substrates and the ATPase activity of Lon (La) proteases are stimulated by unstructured proteins such as alpha-casein. This stimulation reveals the simultaneous interaction of Lon with two proteolytic substrates--alpha-casein and the peptide substrate. To understand the cellular function of this stimulation, it is important to determine the physical properties of Lon stimulators. The abilities of compositionally simple random copolymers of amino acids (rcAAs) to stimulate the peptidase and ATPase activities of the Lon protease from Mycobacterium smegmatis (Ms-Lon) and its N-terminal truncation mutant (N-E226) were determined. We report that cationic but not anionic rcAAs stimulated Ms-Lon's peptidase activity but were themselves poor substrates for the enzyme. Peptidase stimulation by rcAAs correlated approximately with the degree of hydrophobicity of these polypeptides and reached levels >10-fold higher than observed previously for Ms-Lon stimulators such as alpha-casein. In contrast to alpha-casein, which stimulates Ms-Lon's peptidase activity by 40% and ATPase activity by 150%, rcAAs stimulated peptidase activity without concomitant stimulation of ATPase activity. Active site labeling experiments suggested that both rcAAs and ATP increased peptidase activity by increasing accessibility to the peptidase active site. Peptidase activity assays in the presence of both alpha-casein and rcAAs revealed that interactions of rcAAs and alpha-casein with Ms-Lon are extremely complex and not mutually exclusive. Specifically, (1) additions of low concentrations of alpha casein (<50 microg/mL) caused a further stimulation of Ms-Lon's rcAA-stimulated peptidase activity; (2) additions of higher concentrations of alpha-casein inhibited Ms-Lon's rcAA-stimulated peptidase activity; (3) additions of all concentrations of alpha-casein inhibited N-E226's rcAA-stimulated peptidase activity. We conclude the Ms-Lon can interact with an rcAA, alpha-casein, and a substrate peptide simultaneously, and that formation of this quaternary complex requires the N-terminal domain of Ms-Lon. These data support models of Ms-Lon that include two allosteric polypeptide binding sites distinct from the catalytic peptidase site. PMID- 11045627 TI - Insights into nucleotide binding in protein kinase A using fluorescent adenosine derivatives. AB - The binding of the methylanthraniloyl derivatives of ATP (mant-ATP), ADP (mant ADP), 2'deoxyATP (mant-2'deoxyATP), and 3'deoxyATP (mant-3'deoxyATP) to the catalytic subunit of protein kinase A was studied to gain insights into the mechanism of nucleotide binding. The binding of the mant nucleotides leads to a large increase in fluorescence energy transfer at 440 nm, allowing direct measurements of nucleotide affinity. The dissociation constant of mant-ADP is identical to that for ADP, while that for mant-ATP is approximately threefold higher than that for ATP. The dissociation constant for mant-3'deoxyATP is approximately fivefold higher than that for 3'deoxyATP while derivatization of 2'deoxyATP does not affect affinity. The time-dependent binding of mant-ATP, mant 2'deoxyATP, and mant-ADP, measured using stopped-flow fluorescence spectroscopy, is best fit to three exponentials. The fast phase is ligand dependent, while the two slower phases are ligand independent. The slower phases are similar but not identical in rate, and have opposite fluorescence amplitudes. Both isomers of mant-ATP are equivalent substrates, as judged by reversed-phase chromatography, although the rate of phosphorylation is approximately 20-fold lower than the natural nucleotide. The kinetic data are consistent with a three-step binding mechanism in which initial association of the nucleotide derivatives produces a highly fluorescent complex. Either one or two conformational changes can occur after the formation of this binary species, but one of the isomerized forms must have low fluorescence compared to the initial binary complex. These data soundly attest to the structural plasticity within the kinase core that may be essential for catalysis. Overall, the mant nucleotides present a useful reporter system for gauging these conformational changes in light of the prevailing three-dimensional models for the enzyme. PMID- 11045628 TI - Redox-related conformational changes in Rhodobacter capsulatus cytochrome c2. AB - WEFT-NOESY and transfer WEFT-NOESY NMR spectra were used to determine the heme proton assignments for Rhodobacter capsulatus ferricytochrome c2. The Fermi contact and pseudo-contact contributions to the paramagnetic effect of the unpaired electron in the oxidized state were evaluated for the heme and ligand protons. The chemical shift assignments for the 1H and 15N NMR spectra were obtained by a combination of 1H-1H and 1H-15N two-dimensional NMR spectroscopy. The short-range nuclear Overhauser effect (NOE) data are consistent with the view that the secondary structure for the oxidized state of this protein closely approximates that of the reduced form, but with redox-related conformational changes between the two redox states. To understand the decrease in stability of the oxidized state of this cytochrome c2 compared to the reduced form, the structural difference between the two redox states were analyzed by the differences in the NOE intensities, pseudo-contact shifts and the hydrogen deuterium exchange rates of the amide protons. We find that the major difference between redox states, although subtle, involve heme protein interactions, orientation of the heme ligands, differences in hydrogen bond networks and, possible alterations in the position of some internal water molecules. Thus, it appears that the general destabilization of cytochrome c2, which occurs on oxidation, is consistent with the alteration of hydrogen bonds that result in changes in the internal dynamics of the protein. PMID- 11045630 TI - Structure of streptococcal pyrogenic exotoxin A reveals a novel metal cluster. AB - The streptococcal pyrogenic toxins A, B, and C (SPEA, SPEB, and SPEC) are responsible for the fever, rash, and other toxicities associated with scarlet fever and streptococcal toxic shock syndrome. This role, together with the ubiquity of diseases caused by Streptococcus pyogenes, have prompted structural analyses of SPEA by several groups. Papageorgiou et al. (1999) have recently reported the structure of SPEA crystallized in the absence of zinc. Zinc has been shown to be important in the ability of some staphylococcal and streptococcal toxins to stimulate proliferation of CD4+ T-cells. Since cadmium is more electron dense than zinc and typically binds interchangeably, we grew crystals in the presence of 10 mM CdCl2. Crystals have been obtained in three space groups, and the structure in the P2(1)2(1)2(1) crystal form has been refined to 1.9 A resolution. The structural analysis revealed an identical tetramer as well as a novel tetrahedral cluster of cadmium in all three crystal forms on a disulfide loop encompassing residues 87-98. No cadmium was bound at the site homologous to the zinc site in staphylococcal enterotoxins C (SECs) despite the high structural homology between SPEA and SECs. Subsequent soaking of crystals grown in the presence of cadmium in 10 mM ZnCl2 showed that zinc binds in this site (indicating it can discriminate between zinc and cadmium ions) using the three ligands (Asp77, His106, and His110) homologous to the SECs plus a fourth ligand (Glu33). PMID- 11045629 TI - Structure-based prediction of binding peptides to MHC class I molecules: application to a broad range of MHC alleles. AB - Specific binding of antigenic peptides to major histocompatibility complex (MHC) class I molecules is a prerequisite for their recognition by cytotoxic T-cells. Prediction of MHC-binding peptides must therefore be incorporated in any predictive algorithm attempting to identify immunodominant T-cell epitopes, based on the amino acid sequence of the protein antigen. Development of predictive algorithms based on experimental binding data requires experimental testing of a very large number of peptides. A complementary approach relies on the structural conservation observed in crystallographically solved peptide-MHC complexes. By this approach, the peptide structure in the MHC groove is used as a template upon which peptide candidates are threaded, and their compatibility to bind is evaluated by statistical pairwise potentials. Our original algorithm based on this approach used the pairwise potential table of Miyazawa and Jernigan (Miyazawa S, Jernigan RL, 1996, J Mol Biol 256:623-644) and succeeded to correctly identify good binders only for MHC molecules with hydrophobic binding pockets, probably because of the high emphasis of hydrophobic interactions in this table. A recently developed pairwise potential table by Betancourt and Thirumalai (Betancourt MR, Thirumalai D, 1999, Protein Sci 8:361-369) that is based on the Miyazawa and Jernigan table describes the hydrophilic interactions more appropriately. In this paper, we demonstrate how the use of this table, together with a new definition of MHC contact residues by which only residues that contribute exclusively to sequence specific binding are included, allows the development of an improved algorithm that can be applied to a wide range of MHC class I alleles. PMID- 11045631 TI - Change in dimerization mode by removal of a single unsatisfied polar residue located at the interface. AB - The importance of unsatisfied hydrogen bonding potential on protein-protein interaction was studied. Two alternate modes of dimerization (conventional and flipped form) of an immunoglobulin light chain variable domain (V(L)) were previously identified. In the flipped form, interface residue Gln89 would have an unsatisfied hydrogen bonding potential. Removal of this Gln should render the flipped dimer as the more favorable quaternary form. High resolution crystallographic studies of the Q89A and Q89L mutants show, as we predicted, that these proteins indeed form flipped dimers with very similar interfaces. A small cavity is present in the Q89A mutant that is reflected in the approximately 100 times lower association constant than found for the Q89L mutant. The association constant of Q89A and Q89L proteins (4 x 10(6) M(-1) and >10(8) M(-1)) are 10- and 1,000-fold higher than that of the wild-type protein that forms conventional dimers clearly showing the energetic reasons for the flipped dimer formation. PMID- 11045632 TI - Life-supporting human complement regulator decay accelerating factor transgenic pig liver xenograft maintains the metabolic function and coagulation in the nonhuman primate for up to 8 days. AB - BACKGROUND: It is not known whether the pig liver is capable of functioning efficiently when transplanted into a primate, neither is there experience in transplanting a liver from a transgenic pigs expressing the human complement regulator human complement regulator decay accelerating factor (h-DAF) into a baboon. The objective of this study was to determine whether the porcine liver would support the metabolic functions of non-human primates and to establish the effect of hDAF expression in the prevention of hyperacute rejection of porcine livers transplanted into primates. METHODS: Five orthotopic liver xenotransplants from pig to baboon were carried out: three from unmodified pigs and two using livers from h-DAF transgenic pigs. FINDINGS: The three control animals transplanted with livers from unmodified pigs survived for less than 12 hr. Baboons transplanted with livers from h-DAF transgenic pigs survived for 4 and 8 days. Hyperacute rejection was not detected in the baboons transplanted with hDAF transgenic pig livers; however, it was demonstrated in the three transplants from unmodified pigs. Baboons transplanted with livers from h-DAF transgenic pigs were extubated at postoperative day 1 and were awake and able to eat and drink. In the recipients of hDAF transgenic pig livers the clotting parameters reached nearly normal levels at day 2 after transplantation and remained normal up to the end of the experiments. In these hDAF liver recipients, porcine fibrinogen was first detected in the baboon plasma 2 hr postreperfusion, and was present up to the end of the experiments. One animal was euthanized at day 8 after development of sepsis and coagulopathy, the other animal arrested at day 4, after an episode of vomiting and aspiration. The postmortem examination of the hDAF transgenic liver xenografts did not demonstrate rejection. INTERPRETATION: The livers from h-DAF transgenic pigs did not undergo hyperacute rejection after orthotopic xenotransplantation in baboons. When HAR is abrogated, the porcine liver maintains sufficient coagulation and protein levels in the baboon up to 8 days after OLT. PMID- 11045633 TI - Matrix metalloproteinases correlate with alveolar-capillary permeability alteration in lung ischemia-reperfusion injury. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are able to degrade the endothelial basal lamina and increase vascular permeability. METHODS: In a porcine model of isolated-reperfused lung, we studied the alveolar-capillary permeability and the zymographic expression of MMP-9 and MMP-2 in the bronchoalveolar lavage fluid of lungs submitted ex vivo to ischemia in three preservation solutions [modified Euro-Collins (EC), low-potassium-dextran, modified-blood]. Twenty-two pigs were randomly divided into three groups according to the preservation solution used. One lung of each pig was rapidly reperfused and analyzed (control lung) although the other lung was reperfused and analyzed after 8 hr of ischemia (ischemic lung). RESULTS: Alveolar-capillary permeability, evaluated by the transferrin leak index, was increased after 8 hr of ischemia compared with controls in the three groups, but was significantly higher in the modified EC group. In the EC group, after 8 hr of ischemia, both proMMP-9 and MMP-9 increased significantly (8.8- and 22-fold, respectively) compared with controls and this increase correlated with the transferrin leak index. Neither proMMP-9 nor MMP-9 increased with the other two preservation solutions. The MMP-2 increase after ischemia was smaller and was also restricted to the EC group. CONCLUSION: MMP expression is enhanced during lung ischemia-reperfusion, especially in the presence of EC and this phenomenon correlates with the alteration of alveolar-capillary permeability. PMID- 11045634 TI - Donor T cells are not required for induction of allograft tolerance in mice treated with antilymphocyte serum, rapamycin, and donor bone marrow cells. AB - BACKGROUND: Postgraft infusion of donor bone marrow cells (BMC) effectively induces tolerance to skin allografts in antilymphocyte serum- and rapamycin treated recipients in fully major histocompatibility complex-mismatched mouse strain combinations. We used various gene knockout mice to examine the role of donor T cells and B cells in BMC-induced allograft tolerance. METHODS: All recipient mice received ALS on days -1 and 2 and rapamycin (6 mg/kg) on day 7 relative to fully major histocompatibility complex-mismatched skin grafting on day 0. Donor BMC prepared either from mice lacking CD4- and/or CD8a-, or CD3epsilon-expressing cells or B cells, or from corresponding wildtype mice, were given on day 7. The level and phenotypes of chimerism was determined by flow cytometry. RESULTS: All T cell- and B cell-deficient BMC were as effective as wild-type BMC in inducing prolongation of skin graft survival. A low degree of chimerism without donor type T cells was detected in tolerant mice given T cell deficient BMC or wild-type BMC 60 days after transplantation. Chimeric cells were composed of B cells and macrophages/monocytes. Low level chimerism without donor T or B cells was also present in tolerant mice given B cell-deficient BMC. CONCLUSION: Donor type T cells and T cell chimerism are not required for induction of allograft tolerance by the antilymphocyte serum/rapamycin/donor BMC infusion protocol. Donor B cells also do not participate in tolerance induction. Thus, infusion of T cell-depleted BMC in conjunction with conventional immunosuppressive regimens will be a simple, safe, and effective way to induce allograft tolerance in clinical organ transplantation. PMID- 11045635 TI - Transmission of porcine endogenous retroviruses in severe combined immunodeficient mice xenotransplanted with fetal porcine pancreatic cells. AB - BACKGROUND: Xenotransplantation using pig organs or tissues may alleviate the human donor organ shortage. However, one concern is the potential transmission of pig pathogens to humans, especially pig endogenous retroviruses (PERV), which infect human cell lines in vitro. In this report, the cross-species in vivo transmission of PERV by xenotransplantation was studied using a severe combined immunodeficient (SCID) mouse model. METHODS: Twenty-one SCID mice were transplanted with fetal pig pancreatic cells and left for periods from three to 41 weeks before being killed. DNA and RNA were extracted from liver, spleen, and brain of these mice, and examined for PERV using nested polymerase chain reaction (PCR) and reverse transcriptase-PCR. The pig mitochondrial cytochrome oxidase II subunit gene (COII) was also amplified to monitor the presence of pig cell microchimerism in xenotransplanted tissues, and a housekeeping gene was included to monitor the DNA quality and quantity. RESULTS: Examination of 39 DNA samples from tissues of the 21 xenografted mice identified two mouse tissues (M4-liver and M19-spleen) that were positive for PERV but negative for COII. A total of 23 (59%) of the mouse tissues were positive for both PERV and COII, 6 (16%) were negative for both, and 8 (20%) were positive for COII only. PCR and direct sequencing of the PCR products identified three PERV variants, which were different from the PERV sequence detected by PCR direct sequencing from the pig donor cells. CONCLUSIONS: The PERV+/COII- results from M4-liver and M19-spleen indicated the presence of PERV transmission from pig to mouse tissue. The PERV variants detected in the mouse tissues indicated that different PERVs were transmissible from the pig to mouse tissue during xenotransplantation. The negative reverse transcriptase-PCR results for PERV from three mouse samples including M4-liver and M19-spleen suggest there was no active PERV transcription in the mouse tissues, although this would need to be studied further. PMID- 11045636 TI - Modulation of pulmonary NA+ pump gene expression during cold storage and reperfusion. AB - BACKGROUND: Reperfusion injury with pulmonary edema continues to be a major complication after lung transplantation. Alveolar fluid homeostasis is regulated by Na+/K+-ATPase activity on the basolateral surface of alveolar epithelial cells. Intact Na+/K+-ATPase is essential to the resolution of pulmonary edema. We characterized the effects of cold ischemia and reperfusion on expression of Na+/K+-ATPase mRNA and protein. METHODS: Baseline values for Na+/K+-ATPase mRNA and protein were determined from freshly harvested lungs with no cold storage time or reperfusion (group I). Group II lungs were analyzed after cold storage times of 12 or 24 hr without subsequent reperfusion. Group III lungs were analyzed after cold storage times of 12 or 24 hr with subsequent reperfusion. Lungs were flushed with either Euro-Collins (EC) or University of Wisconsin (UW) solution in each group. All samples were quantified for Na+/K+-ATPase mRNA and Na+/K+-ATPase protein. Physiological parameters including oxygenation and compliance were also measured. RESULTS: There were no significant differences in the level of mRNA and protein for samples that were cold stored without reperfusion (group II). With reperfusion (group III) there was a significant increase in the level of the Na+/K+-ATPase mRNA after 12 hr of storage for both EC and UW. After 24 hr of storage and subsequent reperfusion, lungs flushed with EC had significantly decreased Na+/K+-ATPase protein and mRNA, although lungs preserved with UW maintained their increased levels of Na+/K+-ATPase protein and mRNA. CONCLUSIONS: Our data suggest that ischemia-reperfusion injury results in an initial up-regulation of Na+/K+-ATPase mRNA. With prolonged injury in lungs preserved with EC, the level of the mRNA decreased with a corresponding decrease in the Na+/K+-ATPase protein. The different response seen in EC versus UW may be explained by better preservation of pump function with UW than EC and correlates with improved physiological function in lungs preserved with UW solution. PMID- 11045637 TI - Pharmacologic graft protection without donor pretreatment in liver transplantation from non-heart-beating donors. AB - BACKGROUND: Non-heart-beating donors (NHBDs) are considered potential sources of transplant organs in an effort to alleviate the problem of donor shortage in clinical liver transplantation. We investigated the possibility of pharmacologic protection of hepatic allograft function from NHBDs without donor pretreatment. METHODS: Orthotopic liver transplantation was performed using pigs. In donors, cardiac arrest was induced by stopping the respirator. Forty-five minutes after cessation of the respirator, the liver was flushed with cold lactated Ringer's solution including heparin and with the University of Wisconsin (UW) solution, and then preserved for 8 hr at 4 degrees C in the UW solution. The pigs were divided into two groups: a control group and a treated group. In the treated group, an endothelin antagonist TAK-044 was added to the UW solutions (10 mg/L), and TAK-044 (10 mg/kg body weight) and a platelet activating factor antagonist E5880 (0.3 mg/kg body weight) were also administered to the recipients. RESULTS: TAK-044 and E5880 treatment significantly increased the 7-day survival rate of the recipients (100% vs. 17%, P<0.05). In the treated group, portal venous pressure immediately after reperfusion of the graft was significantly lower than in the control group, and postoperative increase in serum concentrations of glutamic oxaloacetic transaminase and total bilirubin was attenuated. Moreover, the energy charge and adenosine triphosphate concentration of the liver were rapidly restored after reperfusion. CONCLUSIONS: Pharmacologic modulation with TAK-044 and E5880 avoiding donor pretreatment can improve the viability of hepatic allografts procured from NHBDs. PMID- 11045638 TI - Leukemia inhibitory factor gene improves skin allograft survival in the mouse model. AB - BACKGROUND: Leukemia inhibitory factor (LIF) is a widely expressed cytokine involved in both local and systemic immune response. Furthermore, it has been implicated in various immunological processes including thymic T cell maturation and embryo implantation. We investigated implication of various modalities in the application of prolonged and viable allograft to the wound, using cytokines and growth factors. MATERIALS: BALB/c and B6D2F1 strains of mice were used either as a skin graft donor or host. LIF cDNA inserted in plasmid vector or the vector alone was injected intradermally in graft skin and observed up to 21 days. LIF, LIF-receptor, gp130, as well as type 1 and 2 T helper cytokine expressions were investigated by reverse transcription polymerasse chain reaction, in situ hybridization, and histological studies. RESULTS: LIF cDNA-treated groups showed significantly improved graft survival compared to the vector-treated control in 21 days postoperatively for grafting from B6D2F1 to BALB/c and BALB/c to B6D2F1. LIF and LIF receptor mRNA expressions were observed 24 hr and 21 days posttransplantation. The gp130 expression was only observed in LIF-treated B6D2F1 to BALB/c allografting on day 21 posttransplantation. LIF transcripts were strongly present in the epidermal, dermal, and subdermal tissues as determined by an in situ hybridization of LIF-treated grafting. CONCLUSIONS: These results suggest that LIF cDNA treatment is an effective and beneficial adjuvant for the skin allograft survival. Improved skin allograft modulation by cytokine gene transfer is a potentially promising therapy for temporary large skin coverage. PMID- 11045639 TI - Influence of donor age on bovine pancreatic islet isolation. AB - BACKGROUND: Pancreatic islets from pigs are largely used for experimental studies. However, pancreas harvesting requires modification of conventional slaughtering to reduce ischemia time. It has been shown that bovine pancreatic islets can be more easily obtained and they show satisfactory in vitro and in vivo function. To improve the isolation procedure we compared the effect of bovine donor age on islet isolation. METHODS: Islets were isolated by collagenase digestion and sequential sieving from calves (6 months of age) and from adult bovine (> 16 months of age). After isolation the number of islet equivalents was calculated and histological and immunohistochemical studies performed. The purity and viability of islet for each preparation was also estimated. In vitro function of islets was evaluated by static insulin secretion assay, and alginate encapsulated islets were transplanted in streptozotocin-induced diabetic rats for in vivo functional evaluation. RESULTS: A significantly higher number of islets were obtained from calf pancreas, compared with adult bovine pancreas. Hystological examination showed intact morphologic features of islets. The purity of islet preparations was higher from calf pancreas than from adult pancreas. Cell viability, and insulin production in presence of high glucose concentration, were not affected by donor age. All animals receiving microencapsulated islets from calves showed normoglycemia for prolonged periods (17-40 days). CONCLUSIONS: These results indicate that pancreatic islet isolation is more efficient from juvenile bovine than from adult. Calf pancreas is a good and convenient source of tissue for massive islet isolation for experimental studies. PMID- 11045640 TI - Pharmacodynamics of mycophenolic acid in heart allograft recipients: correlation of lymphocyte proliferation and activation with pharmacokinetics and graft histology. AB - BACKGROUND: Assays of drug blood levels are used for therapeutic immunosuppressive drug monitoring (pharmacokinetics, PK). We monitored lymphocyte functions (pharmacodynamics, PD) in allograft recipients treated with mycophenolic acid (MPA) to determine its mechanisms and the relationships among dose levels, PK, PD, and histological severity of graft rejection. METHODS: Lewis rats transplanted with Brown Norway (BN) rat hearts were treated with different dose levels of MPA for 8, 15, or 29 days at which times grafts were removed and scored for rejection grade. Blood was analyzed (high-performance liquid chromatography) for MPA plasma concentrations (area under the concentration-time curve0-24 hr, C6 hr, trough) and for lymphocyte functions using concanavalin A stimulated whole blood assays to measure lymphocyte proliferation (tritium labeled thymidine incorporation and flow cytometric bivariate proliferating nuclear cell antigen/DNA analysis) and activation (percent lymphocytes expressing CD25 or CD134). PD values were AUE0-24 hr (area under the PD effect-time curve), maximum inhibition and trough. RESULTS: MPA equipotently suppressed (by flow cytometry) both proliferation and activation and these effects correlated with MPA plasma levels (r2=0.80-0.91). Relationships among MPA dose levels, PK and PD were clear, direct, and reproducible. Correlation coefficients after 8 days of MPA treatment were: 0.90, 0.87, and 0.49 for MPA PK (AUC0-24 hr, C6 hr and trough) versus rejection scores; 0.80-0.89, 0.86-0.92, and 0.25-0.52 for PD flow cytometric assays (AUE0-24 hr, maximum inhibition, and trough) versus rejection scores. CONCLUSIONS: MPA inhibits both lymphocyte proliferation and activation. PD by flow cytometry (FCM) correlates highly with severity of graft rejection, showing that PD of MPA measured in peripheral blood predicts immune cell activity in graft tissue. PMID- 11045641 TI - Peripheral vascular occlusive disease in renal transplant recipients: risk factors and impact on kidney allograft survival. AB - BACKGROUND: This study evaluated the relationship between renal transplantation and the evolution of lower extremity peripheral vascular occlusive disease (PVOD). METHODS: A total of 664 adult renal allograft recipients from 1985-1995 were retrospectively reviewed for atherosclerotic risk factors and peripheral vascular occlusive disease (PVOD). PVOD events were defined as bypass, major amputation, claudication, or percutaneous angioplasty. Follow-up ranged from 2-12 years. RESULTS: The cumulative 5- and 10-year incidences of lower extremity PVOD after renal transplantation were 4.2 and 5.9%. Eight of 14 patients (57%) with pretransplant PVOD had additional PVOD events versus de novo appearance of PVOD in 21/650 patients (3.2%; P<0.0001). In a proportional hazards model, age, preoperative PVOD, diabetes, and postoperative smoking were independent risk factors for the development of PVOD after transplantation. Recipients with lower extremity PVOD had significantly lower 10-year patient and graft survival. Increased graft failure was due to an excess of deaths with a functioning graft. A total of 34 major interventions were performed. One- and two-year limb salvage rates were 64.2 and 53.8%. CONCLUSIONS: Lower extremity PVOD after renal transplantation is associated with diminished patient survival, and affects kidney graft survival via disproportionate patient attrition. Age, preoperative PVOD, diabetes, and postoperative smoking are important risk factors. Transplantation does not appear to either accelerate or retard the progression of disease. An aggressive approach towards limb salvage in properly selected patients is justifiable. PMID- 11045642 TI - Flow cytometric detection of HLA-specific antibodies as a predictor of heart allograft rejection. AB - BACKGROUND: Historically, panel reactive antibody (PRA) analysis to detect HLA antibodies has been performed using cell-based complement-dependent cytotoxicity (CDC) techniques. Recently, a flow cytometric procedure (FlowPRA) was introduced as an alternative approach to detect HLA antibodies. The flow methodology, using a solid phase matrix to which soluble HLA class I or class II antigens are attached is significantly more sensitive than CDC assays. However, the clinical relevance of antibodies detected exclusively by FlowPRAhas not been established. In this study of cardiac allograft recipients, FlowPRA was performed on pretransplant sera with no detectable PRA activity as assessed by CDC assays. FlowPRA antibody activity was then correlated with clinical outcome. METHODS: PRA analysis by anti-human globulin enhanced (AHG) CDC and FlowPRA was performed on sera corresponding to final cross-match specimens from 219 cardiac allograft recipients. In addition, sera collected 3-6 months posttransplant from 91 patients were evaluated. The presence or absence of antibodies was correlated with episodes of rejection and patient survival. A rejection episode was considered to have occurred based on treatment with antirejection medication and/or histology. RESULTS: By CDC, 12 patients (5.5%) had pretransplant PRA >10%. In contrast, 72 patients (32.9%) had pretransplant anti-HLA antibodies detectable by FlowPRA (34 patients with only class I antibodies; 7 patients with only class II antibodies; 31 patients with both class I and class II antibodies). A highly significant association (P<0.001) was observed between pretransplant HLA antibodies detected by FlowPRA and episodes of rejection that occurred during the first posttransplant year. Fifteen patients died within the first year posttransplant. Of nine retrospective flow cytometric cross-matches that were performed, two were in recipients who had no pretransplant antibodies detectable by FlowPRA. Both of these cross-matches were negative. In contrast, five of seven cross-matches were positive among recipients who had FlowPRA detectable pretransplant antibodies. Posttransplant serum specimens from 91 patients were also assessed for antibodies by FlowPRA. Among this group, 58 patients had FlowPRA antibodies and there was a trend (although not statistically significant) for a biopsy documented episode of rejection to have occurred among patients with these antibodies. CONCLUSIONS: Collectively, our data suggest that pre- and posttransplant HLA antibodies detectable by FlowPRA and not AHG-CDC identify cardiac allograft recipients at risk for rejection. Furthermore, a positive donor reactive flow cytometric cross-match is significantly associated with graft loss. Thus, we believe that detection and identification of HLA-specific antibodies can be used to stratify patients into high and low risk categories. An important observation of this study is that in the majority of donor:recipient pairs, pretransplant HLA antibodies were not directed against donor antigens. We speculate that these non-donor-directed antibodies are surrogate markers that correspond to previous T cell activation. Thus, the rejection episodes that occur in these patients are in response to donor-derived MHC peptides that share cryptic determinants with the HLA antigens that initially sensitized the patient. PMID- 11045643 TI - Use of allochimeric proteins to mitigate graft-versus-host and host-versus-graft immune responses to rat small bowel allografts. AB - BACKGROUND: We aimed to identify the polymorphic epitopes that mitigate graft versus-host disease (GvHD) and host-versus-graft response (HvGR) toward rat small bowel allografts in rats. METHODS: We tailored class I major histocompatibility complex (MHC) allochimeric antigens encoding 10 al-helical (alpha(1h)l58-80 RT1.Aa) or 4 (alpha(1h)l/u62-69-RT1.Aa) polymorphic amino acids. In the GvHD model, ACI (RT1a) donors were pretreated (day -14) with an intrathymic injection of alpha(1h)l58-80-RT1.Aa, alpha(1h)l/u62-69-RT1.Aa, or RT1.Al protein, with or without simultaneous intravenous injection of anti-T-cell receptor R73 monoclonal antibodies. Wistar-Furth (WF; RT1u) donors were tested with a similar protocol. In the HvGR model, ACI recipients were treated with a protocol designed to induce transplantation tolerance toward WF heart allografts: a portal vein injection of alpha(1h)l/u62-69-RT1.Aa protein and cyclosporine (4 mg/kg, intramuscular; days 0 6). RESULTS: GvHD was prevented in all (ACI x LEW) F1 recipients (RT1a/l) by pretreating ACI donors with R73 monoclonal antibody and recipient RT1.Al or alpha(1h)l58-80-RT1.Aa protein. Similarly, pretreatment of WF donors with RT1.Aa protein also prevented GvHD in (ACI x WF) F1 recipients. However, in a combined GvHD/HvGR model, ACI recipient perioperative treatment designed to prevent HvGR only modestly prolonged WF small bowel allograft survival (27.7+/-5.3 days compared to 17.4+/-4.6 days in the cyclosporine-alone group). In contrast, application of the two protocols significantly prolonged WF allograft survival (55.6+/-34.6 days), with two of seven recipients surviving more than 100 days. CONCLUSION: Simultaneous inhibition of GvHD and HvGR significantly prolongs small bowel allograft survival. PMID- 11045644 TI - A differential requirement for CD8+ donor cells in the augmentation of allograft survival by posttransplantation administration of donor spleen cells and donor bone marrow cells. AB - BACKGROUND: Peritransplant treatment with antithymocyte serum (ATS) and posttransplantation administration of donor bone marrow or donor splenocytes results in extended skin allograft survival. In this study, we examined the molecular basis of the tolerance promoting effect of donor bone marrow (BMC) cells and splenocytes with emphasis on the role of CD8 expression on the donor cells. METHODS: (C57BL/6J x A/J)F1 mice were treated on days -1 and +2 with ATS relative to transplantation with C3H/HeJ skin. On day +7, they were infused with CD8+ BMC, CD8- BMC, CD8+ splenocytes, or CD8- splenocyte donor subpopulations isolated by magnetic or fluorescence-based sorting. In additional experiments, B10.D2(R107) mice were treated in the same manner with C57BL/6 skin and BMC or splenocytes from C57BL/6 mice in which the CD8alpha gene had been inactivated. RESULTS: CD8+ donor bone marrow cells induced operational tolerance (defined as graft acceptance in the absence of chronic immunosuppression) in skin graft recipients at a dose that was reduced by 250-fold relative to unfractionated bone marrow cells (1.0x10(5) cells per recipient, median survival time (MST)=41 days vs. 2.5x10(7) cells per recipient, MST=49 days, P=0.40). Similarly, donor bone marrow cells from CD8 knockout mice did not promote graft acceptance (MST=98 days vs. animals not treated with bone marrow cells, MST=70 days, P=0.16). In contrast, the extension of graft survival by donor splenocytes did not require the presence of CD8+ donor cells because splenocytes depleted of CD8+ cells extended graft survival (MST=55 days) as well as unsorted splenocytes (44 days, P=0.2), and splenocytes from CD8 knockout animals (MST=145 days) extended graft survival at least as well as unsorted splenocytes (MST=74 days, P=0.4) CONCLUSIONS: These results suggest that the prolongation of graft survival by donor bone marrow is dependent on the presence of the CD8 molecule, whereas prolongation by donor splenocytes is not. Therefore, we suggest that the prolongation of graft survival by these cell types occurs via distinct molecular mechanisms probably mediated by different cell types. PMID- 11045645 TI - Cytokine and cytotoxic molecule gene expression determined in peripheral blood mononuclear cells in the diagnosis of acute renal rejection. AB - BACKGROUND: Prevention of acute rejection is the most prevalent measure used to reduce the long-term risk of chronic allograft rejection. Until now, biopsy was the only useful diagnostic tool for monitoring allograft acute rejection, but invasiveness of this procedure limits its use. The aim of this study was to investigate the implication of peripheral blood immune markers as a predictive diagnostic tool preceding biopsy in acute renal allograft rejection determination. METHODS: Of the 61 patients studied, 13 had no rejection episodes, 8 had a proven acute rejection, and 40 were excluded for graft dysfunction causes. Mitogen-induced peripheral blood mononuclear cells were tested for interleukin- (IL) 2, IL-4, IL-5, IL-6, IL-10, IL-15, Interferon-gamma, Perforin, Granzyme B, and Fas L using semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). An up-regulated mRNA expression value was calculated in which a patient's sample was deemed positive if its differential expression value was equal or higher than the mean differential expression value calculated from the nonrejecting patients. RESULTS: IL-4, IL-5, IL-6, Interferon-gamma, Perforin, and Granzyme B mRNA levels were associated with acute rejection. When at least two of these cytokine markers were up-regulated in a given patient, 75% of the rejecting recipients were identified against 15% of the nonrejecting patients. CONCLUSIONS: We have shown that acute rejection episodes in renal transplant recipients were associated with an increase in mRNA expression of cytokines in mitogen-induced peripheral blood mononuclear cells. The evaluation of pro inflammatory cytokines and cytotoxic molecules prove useful in the clinical identification of acutely rejecting transplant recipients and in the justification of concomitant antirejection therapy before histological diagnosis confirmation. PMID- 11045646 TI - Cadaveric renal allograft at the time of implantation has the similar immunological features with the rejecting allograft. AB - BACKGROUND: Compared to living donations, cadaveric transplants have a poorer outcome, and the immunologic status of renal tissues at the time of transplantation might influence the final outcome of the renal allograft. PATIENTS AND METHODS: We used quantitative RT-PCR to evaluate the differential expression of cytokine genes from 37 implantation tissues [18 cadaveric tissues (CI), 19 specimens from living donors (LI)]. We compared them with 17 acutely rejecting allograft (AR). RESULTS: Acute rejection within 6 months after transplantation occurred 8 times in patients with cadaveric allograft, but the living-donor recipients experienced 4 episodes (P<0.05). Proinflammatory cytokines were co-expressed more frequently in CI than in LI. The levels of IFN gamma, TNF-alpha and IL-10 mRNA were also higher in CI. We compared the profiles of several cytokine expressions of CI with those of AR. The messages for IL-6 were more abundant in the CI, IFN-gamma was more expressed in AR, and the other cytokine expression levels were similar in both types. However, when comparing LI and AR, all the cytokine messages except IL-6 were up-regulated in AR than in LI. In CI, the levels of cytokine gene expressions were similar despite various cold ischemic time except IL-10 that were elevated for those cases where the operation was done within 4 hr of nephrectomy. CONCLUSIONS: The numbers and levels of gene transcription of inflammatory cytokines were higher in the tissues from a cadaver, and were not different from those of AR. This immunologic hostility at the time of implantation would contribute to the poorer outcome of cadaveric allograft. PMID- 11045647 TI - IgG2 anti-Galalpha1-3Gal does not induce porcine aortic endothelial cell accommodation in vitro. AB - BACKGROUND: Xenografts that have been protected from hyperacute rejection (HAR) are termed accommodated if they are not then rejected despite the presence of xenoantibody. It has been proposed that IgG may confer resistance to complement dependent cytotoxicity (CDC), a conventional in vitro marker of accommodation. We hypothesized that noncytotoxic IgG2 anti-Galalpha1-3Gal was responsible for this effect. METHODS AND RESULTS: We purified IgG anti-Galalpha1-3Gal from pooled human normal immunoglobulin and three sera, by elution from protein G and Galalpha1-3Gal-R immunoadsorbents. The eluates were IgM free and > or =95% IgG2. They bound to Galalpha1-3Gal, porcine aortic endothelial cells (PAEC) and lymphocytes. It was not possible to block IgM binding to PAEC or lymphocytes using IgG anti-Galalpha1-3Gal (200 microg/ml). The eluates were noncytotoxic in micro-CDC assays. To investigate accommodation, PAEC were cultured with subsaturating doses of the four IgG eluates for up to 144 hr. Resistance of nontrypsinized PAEC to CDC by human serum was measured in a cell viability assay. PAEC were not rendered resistant to CDC in any of the experiments. To investigate the possibility that accommodation might be induced by non-Galalpha1-3Gal IgG, the experiments were repeated using HNIg, again with no protection demonstrated. CONCLUSIONS: Using primary PAEC monolayers, we were unable to induce resistance to CDC with human normal immunoglobulin and its IgG2 anti-Gabeta1-3Gal subset. This contradicts previous experiments using trypsinized, immortalized cells. Although resistance to CDC is not an ideal marker of accommodation, the detrimental effects of IgG make it unlikely that it will become a useful clinical means of inducing accommodation. PMID- 11045648 TI - Hydrophobic extracts of a Chinese herb (CMX-13) exhibit potent immunosuppressive properties and prevent acute rejection in a highly histoincompatible model of rat lung transplantation. AB - BACKGROUND: The potential of higher plants as sources for new immunosuppressive medications is well recognized. In our experiments we investigated the immunosuppressive effect of a highly refined and potent extract of a Chinese herbal preparation, CMX-13, on inhibiting acute allograft rejection (AR) in a highly histoincompatible rat lung transplant model, BN-->LEW, and on lymphocyte activation and cytokine gene expression in vitro. METHODS: Left lung transplants: the control group (group 1) received only dimethylsulfoxide (DMSO) which is the solvent for CMX-13. Group 2 received intramuscular cyclosporin A (CsA, 25 mg/kg) on day 2 posttransplant. Group 3 and 4 received i.p. CMX-13 (0.5 mg/day, low dose and 5 mg/day, high dose, respectively) on day 1, 2, and 3 posttransplant. All animals were killed on day 6 posttransplant. Several pathological categories of inflammation were examined. In vitro experiments: rat spleen cells were incubated with Con A or irradiated stimulator cells with/without serial dilutions of CMX-13 or CsA. Cell proliferation was measured by 3H-thymidine incorporation. mRNA expression of interleukin-2 and interferon-gamma was examined by reverse transcriptase-polymerase chain reaction. RESULTS: The severity of AR in animals receiving high dose CMX-13 was significantly reduced (stage II, P<0.05) compared with controls (stage IV). Significant differences were also seen when more specific parameters of inflammation were examined (necrosis, 0 vs. 1.7+/-1.0, P<0.05; interalveolar hemorrhage, 0 vs. 3.0+/-0.9, P<0.05). The responses seen in the animals treated with high dose CMX-13 were similar to those in the CsA group. CMX-13 inhibited T cell proliferative responses induced by Con A and alloantigen stimulation in a dose-dependent manner that were similar to CsA. Interleukin-2, and interferon-gamma mRNA expression in Con A-stimulated spleen cells was not inhibited by CMX-13 although CsA showed significant inhibition. CONCLUSIONS: 1) CMX-13 significantly reduces the stage of AR and parameters of inflammation in a highly histoincompatible rat lung transplant model. 2) CMX-13 has equal potency to CsA in the inhibition of Con A and alloantigen stimulated rat spleen cell proliferation. 3) CMX-13 showed no inhibitory effects on IL-2 and gamma-IFN mRNA expression, suggesting that its mechanism of action is different from CsA. 4) CMX 13 or derivatives may have potential utility as an immunosuppressive agent(s) in modulation of AR and management of other inflammatory and immunological disorders. PMID- 11045649 TI - Increased impact of acute rejection on chronic allograft failure in recent era. AB - BACKGROUND: Acute rejection (AR) remains a major risk factor for the development of chronic renal allograft failure (CAF), which is a major cause of late graft loss. With the introduction of several newer immunosuppressive agents (e.g., mycophenolate mofetil, tacrolimus and neoral) acute rejection rates have been steadily decreasing. However, the incidence of CAF has not decreased as dramatically as the incidence of acute rejection. One possible explanation is that the impact of AR on CAF is changing. The goal of this study was to analyze the relative impact of AR era on the development of CAF. METHODS: We evaluated 63,045 primary renal transplant recipients reported to the USRDS from 1988 to 1997. CAF was defined as graft loss after 6 months posttransplantation, censored for death, acute rejection, thrombosis, infection, surgical complications, or recurrent disease. A Cox proportional hazard model correcting for 15 possible confounding factors evaluated the relative impact of AR on CAF. The era effect (years 1988-1989, 1990-1991, 1992-1993, 1994-1995 and 1996-1997) was evaluated by an era versus AR interaction term. RESULTS: An AR episode within the first 6 months after transplantation was the most important risk factor for subsequent CAF (RR=2.4, CI 2.3-2.5). Compared with the reference group (1988-89 with no rejection), having an AR episode in 1988-89, 1990-1991, 1992-1993, 1994-1995, and 1996-1997, conferred a 1.67, 2.35, 3.4, 4.98 and 5.2-fold relative risk for the subsequent development of CAF (P<0.001). CONCLUSIONS: Independently of known confounding variables, the impact of AR on CAF has significantly increased from 1988 to 1997. This effect may in part explain the relative lack of improvements in long term renal allograft survival, despite a decline in AR rates. PMID- 11045650 TI - Amplification of immunological functions by subcutaneous injection of intermediate-high dose interleukin-2 for 2 years after autologous stem cell transplantation in children with stage IV neuroblastoma. AB - BACKGROUND: Immunotherapy given post-autologous stem cell transplantation may eliminate residual tumor cells escaping the conditioning protocol. METHODS: Five children suffering from stage IV neuroblastoma were treated by recombinant interleukin-2 (IL-2) post-autologous peripheral blood stem cell transplantation. The patients' peripheral mononuclear cells were monitored for CD3+ and CD56+ levels, their proliferative response and killing of various cell lines targets. RESULTS: An increase in the level of total lymphocytes, mainly due to expansion of T cells, and enhanced proliferative response to phytohemaglutinin were observed. Elevated cytotoxicity against K562 and neuroblastoma target cells was detected in four patients and against K562 targets in one patient. Toxicity included mild thrombocytopenia, and fever in four patients and mild to moderate encephalopathy which necessitated withdrawing one patient from the protocol. Three of five patients studied are alive today, one of them whose IL-2 was stopped, is in relapse. Two patients have died. CONCLUSIONS: Immunotherapy with s.c. intermediate-high dose IL-2 is feasible and results in expansion of T cells and in stimulation of killing activity against several targets including in some cases, neuroblastoma tumor cells. PMID- 11045651 TI - Rapamycin induces transforming growth factor-beta production by lymphocytes. AB - BACKGROUND: Under certain conditions rapamycin and transforming growth factor- (TGF) beta have similar immunoregulatory effects, suggesting a potential functional link between rapamycin and TGF-beta. METHODS: Splenic leukocytes were stimulated in vitro with anti-CD3 or with allogeneic cells in vivo in the presence or absence of rapamycin. TGF-beta production by activated lymphocytes was quantitated using ELISA. RESULTS: Splenic leukocytes from BALB/c mice that were primed with allogeneic cells and conditioned with rapamycin in vivo as well as splenic leukocytes that were treated with rapamycin in vitro produced significantly higher levels of TGF-beta upon anti-CD3 stimulation as compared with untreated controls. CONCLUSION: Our data suggest that rapamycin can program activated lymphocytes to produce TGF-beta. Thus, the immunosuppressive effects of rapamycin may be partially mediated by TGF-beta. PMID- 11045652 TI - Increased nuclear factor-kappaB and angiotensinogen gene expression in posttransplant recurrent focal segmental glomerulosclerosis. AB - In an attempt to identify potential markers of steroid-resistance in focal segmental glomerulosclerosis (FSGS) we evaluated intra-graft gene expression of IkappaBalpha, nuclear factor-kappaB (NF-kappaB), and angiotensinogen in 60 biopsies from 27 pediatric renal transplant recipients. Intra-graft NF-kappaB expression was significantly elevated in recurrent FSGS (R-FSGS) (218.3 + 55.6 ag/fg versus NON-FSGS 121.1 + 19.9, P=0.04) but not in acute rejection. NF kappaB:IkappaBalpha ratios were higher in cadaveric donor versus living related donor recipients (15.7 + 2.8 vs. 8.8 + 1.3, respectively, P=0.015), and in African-American versus Caucasian recipients (15.6 + 2.9 vs. 9.1 + 1.3, respectively, P=0.03). Intra-graft angiotensinogen gene expression was significantly elevated in R-FSGS (30.5 + 8.8 ag/fg R-FSGS vs. 16.0 + 4.7 NON FSGS, P=0.009). We conclude that increased NF-kappaB and angiotensinogen gene expression are associated with R-FSGS. Increased NF-kappaB:IkappaBalpha ratios are associated with cadaveric donor recipients and African-American race. PMID- 11045653 TI - Continuous lamivudine treatment in an OLT patient despite the appearance of HBV resistant strain: longitudinal analysis. PMID- 11045654 TI - Xenotransplantation and the Council of Europe. PMID- 11045655 TI - Response to "A Novel United Network for Organ Sharing Region Kidney Allocation Plan Improves Transplant Access for Minority Candidates". PMID- 11045656 TI - Standards for economic and quality of life studies in transplantation. AB - There are unique requirements in conducting and reporting economic and quality of life investigations in medicine as compared with more traditional studies involving clinical outcomes. In addition, there are several unique characteristics of the discipline of transplantation that also bear attention in economic and quality of life studies. To provide guidelines for future research and reporting of future research, a consensus conference of transplant professionals was convened to discuss these issues. Five different areas were addressed: "Conducting an Economic Analysis," "Reporting an Economic Analysis," "Quality of Life Studies in Transplantation," "Ethical and Conflict of Interest Issues Between Sponsors and Investigators," and "Future Directions for Research." A series of recommendations for each of these areas with reference to relevant literature is presented. PMID- 11045657 TI - Estimating the benefits of solitary pancreas transplantation in nonuremic patients with type 1 diabetes mellitus: a theoretical analysis. AB - BACKGROUND: The goal of early pancreas transplantation in type 1 diabetes mellitus is to achieve euglycemia and thereby prevent the renal, retinal, and vascular complications of this disease. The purpose of this analysis was to examine the conditions and assumptions that would make early solitary pancreas a viable therapeutic option. METHODS: A Markov model was constructed to compare outcomes for patients with type 1 diabetes mellitus and early overt nephropathy assigned to either standard insulin therapy or solitary pancreas transplantation. Probabilities for development of end stage renal disease (ESRD), blindness, mortality, and direct health care costs were taken from the literature. Utility scores for the relevant health states were determined by the standard gamble method on 16 type 1 diabetic patients suitable for pancreas transplantation. RESULTS: Assuming a baseline graft life expectancy for the pancreas of 10 years, early pancreas transplantation could provide 0.42 more life years and 2.2 more quality adjusted life years (discounted at 3%) to patients above standard insulin therapy. The model was sensitive to estimates of pancreas graft life expectancy (<8 years being inadequate to extend patient life), timing of pancreas transplantation with respect to rate of progression to ESRD, and drug nephrotoxicity rates. The incremental costs (charges) for early pancreas transplantation over standard therapy were estimated to be modestly high (about $56,600/quality adjusted life year for the baseline case). Pancreas transplant costs were also a very sensitive parameter in the cost utility analysis. CONCLUSIONS: The success of early solitary pancreas transplantation may well be at the stage to consider a trial in selected type 1 diabetic patients at risk for renal and retinal disease. PMID- 11045658 TI - Pathology of ALY mice: congenital immunodeficiency with lymph node and Peyer's patch defects. AB - The ALY-alyl/aly mouse is a new and unique animal model of primary immunodeficiency with autosomal recessive inheritance. The ALY mouse is devoid of superficial and profound lymph nodes and Peyer's patches. Furthermore, the lymphoid follicles and marginal zones are not clearly identified in the spleen. In addition to these structural defects, in the present study, we show that some B subpopulations are defective. Firstly, the thymic B lymphocytes are very rare. Secondly, the B220hi sIghi B subpopulation in the bone marrow is not detected as a clear cluster on FACS analyses. Thirdly, the B220 slg+ cells in the bone marrow are very rare in both ALY-aly/aly and ALY-alyl+ mice. By contrast, the NK activity is normal. Taken together, the ALY mouse is an invaluable model to elucidate the immunological networks between the lymphoid structures (lymph nodes, Peyer's patches, lymphoid follicles, etc.) and functions. PMID- 11045659 TI - Neutrophil adhesion and transmigration through bovine endothelial cells in vitro by protein H and LPS of Pasteurella multocida. AB - This study describes an in vitro investigation on the role of Pasteurella multocida cells and its isolated protein H and LPS on neutrophil adhesion and migration through bovine endothelial cell monolayers. P. multicoda cells, protein H and LPS increased the adhesion and transmigration of neutrophils through BAEC. The bacteria/cell ratio of 100 for P. multocida, protein H concentration 0.05-0.2 microM and LPS concentration 0.5-1.0 microM respectively, induced the maximum adhesion and transmigration of neutrophils through BAEC. The optimal time of incubation with bacteria or bacterial products was 4-6 h. Our results confirm the role of Gram-negative bacteria and of components of the outer membrane such as protein H or LPS in activating the neutrophils and in promoting the adhesion and cells transmigration from the vessels to the site of inflammation. PMID- 11045660 TI - Parenteral administration of an activating monoclonal antibody to the alpha1beta1 integrin in dogs. AB - In mice, monoclonal antibody (mAb) to the alpha1 integrin abrogate gastro intestinal damage during graft-versus-host-disease (GVHD), suggesting anti alpha1 mAb as candidates for treatment in humans as well. Our current data show that one such reagent, mAb 1B3.1, when immobilized to plastic wells via rabbit- anti murine (ram) immunoglobulin (Ig) induces a protein kinase-dependent spreading of activated human T cells. Furthermore, it significantly increases the proliferative response, and expression of interleukin-2 (IL-2) receptors (R) and CD69, of resting T cells, expressing minimal integrin on the cell surface, to sub optimal stimulation by anti-CD3 mAb. We found, in addition, that mAb 1B3.1 a) immuno-precipitates alpha1beta1 integrins from cell-surface iodinated canine epithelial cells b) is highly reactive with canine T cells after their activation and c) inhibits adhesion of canine T cells to collagen IV. Despite the potential ability of the mAb to co-activate T cells in vitro, two dogs that received 4 injections of 0.5-0.3 mg/Kg of mAb 1B3.1 remained healthy, developing only marginal transient lymphopenia. Injection of 0.75mg/Kg in a third dog induced a more marked lymphopenia, and an additional dose of 1.0 mg/Kg 2 weeks later was followed by gastrointestinal hemorrhage. importantly, the lymphopenia was associated with a greater and more persistent decrease of CD8+ than of CD4+ T cells, leading to an increase in the CD4/CD8 ratio 24 hours after the injection. Thus, despite it's co-activating effects in vitro, administration of this mAb in vivo is feasible when appropriately dosed, and may have immuno-modulatory effects. PMID- 11045661 TI - Reduced levels of Hsp70 result in a therapeutic effect of 15-deoxyspergualin on acute graft-versus-host disease in (DA x LEW)F1 rats. AB - We have shown previously that increased levels of hsp70, and antibodies reactive with hsp70 parallel the onset and severity of graft-versus-host disease (GVHD) in a parent --> (DA x LEW)F1 rat model. In this study we have assessed the effect of reducing the levels of the 70 kDa heat shock protein (hsp70), on the morbidity and mortality of acute GVHD in (DA x LEW)F1 rats. The reduction was accomplished by the administration of 15-deoxyspergualin (DSG), an immunosuppressive agent which binds to a constitutively expressed member of the 70 kDa heat shock protein family. DSG administered via three different protocols reduced GVHD-associated morbidity. One of the regimens, which consisted of intermittent DSG administration, also significantly reduced GVHD associated mortality. This DSG treatment reduced hsp70 levels in spleen and lymph nodes, inhibited anti-hsp70 antibody production, and diminished the serum levels of IL-2, IFN-gamma, TNF alpha, and IL-10. IL-4 levels in the serum did not change during GVHD and were not effected by DSG. These results show that the mechanism of DSG immunosuppressive effect in rat GVHD may involve DSG's capacity to bind to hsp70, which in turn may lead to a decrease in levels of circulating anti-hsp70 antibodies, and reduced production of cytokines. PMID- 11045662 TI - Abrogation of negative selection by GVHR induced by minor histocompatibility antigens or H-2D antigen alone. AB - Allogeneic bone marrow chimeras were prepared by donor and recipient combinations that differed in minor histocompatibility loci or H-2D locus alone. When 1 x 10(5) splenic T cells were inoculated in addition to T cell-depleted bone marrow cells (1 x 10(7)), clinically detectable GVHR was induced. In these GVHR chimeras, substantial numbers of T cells reactive to either donor or recipient antigens were both phenotypically and functionally detected. The mechanisms underlying the abrogation of intrathymic negative selection are discussed. PMID- 11045663 TI - Distinct patterns of cytokine gene suppression by the equivalent effective doses of cyclosporine and tacrolimus in rat heart allografts. AB - In vitro studies of the mode of action of cyclosporine (CsA) and tacrolimus have indicated that both drugs produce immunosuppression by a quite similar cellular and molecular mechanism to block T cell receptor emanated transcriptional activation of interleukin(IL)-2 and other cytokine genes. Herein, we show that there are distinct patterns of cytokine gene expression in rat heart allografts under equivalent effective doses ("optimal dose") of CsA and tacrolimus. The optimal doses of CsA (10 mg/kg/day) and tacrolimus (3.2 mg/kg/day), which induce similar mean graft survival time (MST), were administered in LEW recipients with ACI heart grafts from day 0 after grafting until sacrifice. Heart grafts were harvested at days 3, 5, and 7. The expression of various cell surface markers, cytokines, and cytotoxic factors was determined by immunohistology and reverse transcriptase-polymerase chain reaction (RFT-PCR). Cell populations that stained positively in the heart tissues of allograft control increased through day 7 for CD4+ and CD8+ T lymphocytes, NKR-Pla+ natural killer (NK) cells, and ED2+ macrophages. CsA and tacrolimus have comparable activity to block these cell local infiltrations. The mRNA levels of the majority of the factors were dramatically up-regulated in the allografts over time, peaking at day 5. The optimal doses of CsA and tacrolimus had similar inhibitory effects on Th1 type cytokine IL-2 and interferon [INF]-gamma), inflammatory cytokine (IL-1beta and tumor necrosis factor [TNF]-alpha), and cytotoxic factor (granzyme B and perforin) mRNA expression. However, the drugs had different effect on Th2 type cytokines (IL-4 and IL-10). Whereas IL-4 expression was not affected by tacrolimus and was enhanced by CsA, IL-10 expression was more significantly suppressed by tacrolimus than CsA. Differences in the suppression of Th2 type cytokine gene expression indicate that the in vivo molecular networks by which CsA and tacrolimus exert their full immunosuppressive activity are not necessarily the same. PMID- 11045664 TI - B cell-mediated infection of stimulated and unstimulated autologous T lymphocytes with HIV-1: role of complement. AB - In vivo, human immunodeficiency virus type 1 (HIV-1) is opsonized with complement fragments and virus-specific antibodies (Ab). Thus, HIV is able to interact with complement receptor (CR) - and Fc receptor (FcR) - positive cells such as B cells, follicular dendritic cells or macrophages. In this study we demonstrate that the interaction between B cells and HIV has an impact on autologous primary T cell infection in vitro. We confirmed the presence of complement-fragments and virus-specific Ab on serum-treated HIV using a virus-capture assay. In experiments with CR2-specific Ab we showed that the virus/B cell interaction was mainly dependent on CR2. In infection experiments immobilisation of HIV on stimulated tonsil B cells greatly enhanced the infection of interleukin (IL)-2 activated autologous tonsil T cells. Surprisingly, enhancement of T cell infection by B cell-HIV complexes was observed even in the absence of mitogenic stimuli such as PMA and was independent of the addition of exogenous IL-2. Taken together, these results indicate that primary B cells are able to efficiently transmit opsonised HIV to autologous primary T cells and induce a massive enhancement of infection. These in vitro experiments mimic the in vivo situation in the lymphoid tissue and suggest an alternative mechanism for the infection of primary T cells. PMID- 11045665 TI - Apolipoprotein E affects the amount, form, and anatomical distribution of amyloid beta-peptide deposition in homozygous APP(V717F) transgenic mice. AB - Apolipoprotein E (apoE) has been implicated as a risk factor for Alzheimer's disease and in the deposition, fibrillogenesis, and clearance of the amyloid beta peptide (Abeta). To examine the in vivo interactions between apoE and Abeta deposition, we examined 12-month-old transgenic (tg) mice expressing human amyloid precursor protein (APP) with the V717F mutation (APP(V717F) homozygous) on an APOE null background. Elimination of APOE resulted in a redistribution and alteration in the character of Abeta deposition in homozygous APP(V717F) tg mice, with a dramatic reduction in cortical and dentate gyrus deposition, prominent increase in diffuse CA1 and CA3 deposition, and prevention of the formation of thioflavin-S-positive deposits. These alterations in Abeta deposition were not mediated by significant changes in regional APP expression, low-density lipoprotein receptor-related protein expression, or soluble Abeta levels. Thus, apoE in APP(V717F) tg mice not only affects the amount and form of Abeta deposition, but also the anatomical distribution of diffuse Abeta deposits. The APP(V717F) tg mouse can serve as a model to investigate genetic influences on the vulnerability of specific neuroanatomical regions to Abeta deposition. PMID- 11045666 TI - Screening for Charcot-Marie-Tooth type 1A and hereditary neuropathy with liability to pressure palsy in archival nerve biopsy samples by direct-double differential PCR. AB - Chromosomal imbalance of the peripheral myelin protein-22 gene (PMP22) is known to be the most frequent genetic abnormality in Charcot-Marie-Tooth disease type 1 (CMT1) and hereditary neuropathy with liability to pressure palsy (HNPP). We applied a new quantitative PCR method, the direct-double-differential PCR (dddPCR), to the gene dosage determination of PMP22. The method allows the quantification of the PMP22 gene copy number independently from DNA fragmentation, even in highly degraded DNA from up to 12-year-old sural nerve biopsy samples. Chromosomal imbalance of the PMP22 gene, which had been detected by examination of four microsatellites located directly adjacent to the PMP22 gene, between the CMT1A-repetition (CMT1A-REP) elements was reliably confirmed by the dddPCR. Using this method we unexpectedly identified two cases with PMP22 imbalance, although morphologically the neuropathies were of a neuronal or axonal type and not of a demyelinating type as usual. One sural nerve biopsy was from a 58-year-old male diabetes mellitus patient with a disproportionately severe polyneuropathy showing a heterozygous duplication of PMP22. The second biopsy exhibiting a heterozygous deletion of PMP22 was from a 58-year-old female patient with a more axonal than demyelinating type of neuropathy without typical tomaculous changes seemingly altered by exogenous, possibly traumatic factors other than diabetes mellitus. Thus, the dddPCR provides a fast and reliable diagnostic tool for the screening and identification of CMTIA and HNPP cases, which is fast and may be essential even when nerve biopsies show morphologically atypical changes. PMID- 11045667 TI - Tau-negative astrocytic star-like inclusions and coiled bodies in dementia with Lewy bodies. AB - To evaluate glial lesions in cases of dementia with Lewy bodies (DLB), we studied the brains of four patients with DLB. Astrocytic star-like inclusions, which resembled tufted astrocytic fibrillary tangles in shape, were found in the cortex of two of these cases. In addition, coiled bodies were found in the white matter of the cerebrum in two cases. The astrocytic star-like inclusions were immunohistochemically negative for tau protein, ubiquitin and alpha-synuclein. The coiled bodies were immunohistochemically negative for tau protein but immunopositive for ubiquitin and alpha-synuclein. These results suggest that in DLB a primary degenerative process takes place in both glial cells and neurons. PMID- 11045668 TI - Transganglionic gracile response following limb amputation in man. AB - Gracile neuroaxonal dystrophy (NAD) is an distinctive morphological alteration of central projecting axon terminals of dorsal root ganglion neurons. Experimentally, lower limb amputation has been shown to accelerate the formation of gracile NAD, suggesting that the transganglionic response to peripheral axotomy may play a role in its development. To determine if a similar response occurs in the human sensory nervous system following peripheral nerve injury, we have performed postmortem histopathological examinations of the dorsal column nuclei of three patients (aged 15, 55, and 77 years old); all of whom had undergone accidental or therapeutic unilateral limb amputation (1 year, 38 years, and 1 year 8 months prior to death, respectively). In a 15-year-old man who underwent therapeutic leg amputation, the gracile nuclei on the transected side revealed reactive gliosis and many small axonal spheroids. The spheroids and fine neurites were immunolabelled with antibodies for growth-associated protein-43, ubiquitin and neuropeptide Y (NPY). Neither routine histological nor immunohistochemical methods demonstrated comparable changes in the contralateral gracile nucleus. In a 77-year-old man who underwent leg amputation, the gracile nucleus on the amputated side was gliotic and showed several NPY and ubiquitin immunoreactive spheroids, which were not seen in the contralateral non-transected side. A 55-year-old man with a history of accidental arm amputation showed well developed NAD in the cuneate nucleus only on the transected side. This study clearly demonstrates the occurrence of transganglionic response to limb amputation in human dorsal column nuclei. The extent of the regenerative and/or degenerative responses may vary depending on the age of the patient and the time interval following the peripheral axotomy. PMID- 11045669 TI - Phenotypic modulation of smooth muscle cells in human cerebral aneurysmal walls. AB - We used immunohistochemical methods to analyze the phenotypes of smooth muscle cells (SMCs) in human cerebral arteries and aneurysmal walls. Thirty-two aneurysmal walls were studied; 31 aneurysmal walls were resected at operation and 1 aneurysm was obtained at autopsy. Seven control arteries were obtained at autopsy. Semiserial sections were subjected to immunohistochemical staining with antibodies to alpha-smooth muscle actin (alpha-SMA), desmin and smooth muscle myosin heavy chain isoforms: SM1, SM2 and SMemb. In control cerebral arteries, SMCs in the media were strongly immunostained for alpha-SMA, desmin, SM1 and SM2; immunoreactivity for SMemb was faint or weakly positive. SMCs in both non ruptured and ruptured aneurysmal walls showed no staining for desmin; the expression of alpha-SMA was well preserved. Compared with control cerebral arteries, in 4 of 11 non-ruptured aneurysmal walls, the staining intensity of SMCs for SMemb was clearly increased. In ruptured aneurysmal walls, the expression of SM2 was lower than in control cerebral arteries and non-ruptured aneurysmal walls. Our study suggests that the phenotype of SMCs in aneurysmal walls is different from the contractile type in the media of normal cerebral arteries, at least partially changing to the synthetic type in some non-ruptured aneurysms. SMCs in ruptured aneurysmal walls may have lost both phenotypes before rupture. Phenotypic modulation of SMCs in the aneurysmal walls appears to be related to a remodeling of the aneurysmal wall and to a rupture mechanism. PMID- 11045670 TI - Midkine, a new neurotrophic factor, is present in glial cytoplasmic inclusions of multiple system atrophy brains. AB - The glial cytoplasmic inclusion (GCI) is a histological hallmark for multiple system atrophy (MSA): these inclusions are found in oligodendrocytes and consist of abnormal granule-coated fibrils of approximately 24- to 40-nm diameter. To clarify the significance of the presence of midkine (MK) in these GCIs, we carried out immunohistochemical, electron and immunoelectron microscopical, and Western blot analyses of MSA brains using a monoclonal antibody against the C terminal region of human MK. Immunohistochemically, most of the GCIs were intensely stained by the antibody to MK. Electron and immunoelectron microscopy showed that the GCIs were composed of MK-positive granule-coated fibrils that were essential constituents of these inclusions. No significant MK immunoreactivity was observed in oligodendrocytes, astrocytes and neurons of the normal control subjects. The presence of MK in MSA brain but not in normal brain was confirmed by Western blotting. Together with the fact that MK is associated with fetal morphogenesis during the midgestation period, the presence of MK immunoreactivity in oligodendroglial GCIs may suggest the existence of a repair mechanism on the basis of morphogenesis in the degenerated oligodendrocytes themselves as well as the affected neurons and their axons through the oligodendrocyte-axon-neuron relationship. PMID- 11045672 TI - Ectopic expression of telencephalin in brains with holoprosencephaly. AB - Telencephalin (TLN), a telencephalon-specific glycoprotein, is exclusively expressed in neurons of the mammalian telencephalon. In the normally developing human brain, TLN immunoreactivity appeared and increased from 35 gestational weeks (GW) in the temporal cortex, and reached adult level at 5 months of postnatal age, being strong in the molecular layer, and weak in the external and internal granular layers. TLN expression corresponded with the development of neuronal dendrites and synapses. In brains with holoprosencephaly TLN immunoreactivity was already strong from as early as 28 GW. Staining was weak in the molecular layer, but strong in the external sparse and middle cellular layers in most cases. Notably, TLN was abundant in the glomerular structures in the internal pyramidal and multiform layers of fetal brains with alobar holoprosencephaly, which disappeared with increasing age. These results indicate premature and ectopic development of the dendrites and synaptic network in holoprosencephaly. PMID- 11045671 TI - Advanced glycation endproduct-modified superoxide dismutase-1 (SOD1)-positive inclusions are common to familial amyotrophic lateral sclerosis patients with SOD1 gene mutations and transgenic mice expressing human SOD1 with a G85R mutation. AB - To clarify the biological significance of the neuronal Lewy body-like hyaline inclusions and astrocytic hyaline inclusions characteristically found in patients with familial amyotrophic lateral sclerosis with superoxide dismutase-1 (SOD1) gene mutations and in transgenic mice expressing human SOD1 with G85R mutation, the detailed protein composition in both types of inclusions was immunohistochemically analyzed using 45 different antibodies. Both types of inclusions had very strong immunoreactivity for SOD1. The SOD1-positive inclusions in both cell types were also immunoreactive for the insoluble advanced glycation endproducts (AGEs) such as Nepsilon-(carboxymethyl)lysine (CML), pyrraline and pentosidine: both inclusions in both conditions were ultrastructurally composed of the granule-coated fibrils that had immunoreactivities to CML and pyrraline. Both types of inclusions were negative for stress-response proteins (SRPs), 4-hydroxy-2-nonenal (HNE), acrolein, nitric oxide synthases (NOSs) and nitrotyrosine as representative markers of oxidative stress. The neurons and astrocytes of the normal individuals and non-transgenic mice showed no significant immunoreactivity for SOD1, AGEs, SRPs, HNE, acrolein, NOSs or nitrotyrosine. Our results suggest that a portion of the SOD1 composing both type of inclusions, probably toxic mutant SOD1, is modified by the AGEs, and that the formation of the AGE-modified SOD1 is one of the mechanisms responsible for the aggregation involving no significant oxidative mechanisms. PMID- 11045673 TI - Developmental expression of myotonic dystrophy protein kinase in brain and its relevance to clinical phenotype. AB - To investigate the pathophysiologic role of myotonic dystrophy protein kinase (DMPK) in the brain in myotonic dystrophy (MD), the developmental characteristics of DMPK immunoreactivity in the central nervous system and its alteration with disease were studied. Eleven patients' brain with MD (5 congenital form, 6 adult form) were examined by immunohistochemistry using a specific antibody against synthetic DMPK peptides, antipeptide DM1, and compared with 30 control brains, including 16 age-matched controls. In controls, DM1-immunoreactive neurons appeared in the early fetal frontal cortex and cerebellar granule cell layer, persisting through 29 weeks of gestation and then disappearing. In contrast, immunoreactive neurons continued to persist in the cerebral cortex and cerebellar granule cell layer of MD patients. When we counted DM1-immunoreactive neurons, the increase over controls was greater in the congenital form of MD than in the adult form, and was greater in the cerebrum than in the cerebellum in both forms of MD. DM1 immunostaining was predominantly nuclear, mirroring Western blotting of subcellular fractions. Differences in DM1 expression related to development and to the two forms of MD may be closely related to the pathogenesis of mental retardation in this disease. PMID- 11045674 TI - The role of lymphotoxin in pathogenesis of polymyositis. AB - Polymyositis (PM) is a cell-mediated autoimmune disease. Perforin (PF), Fas ligand (FasL) and TNF-alpha are considered to be important factors in cytotoxic T lymphocyte-mediated cell injury, and several studies have established a role of lymphotoxin (LT) in T helper type 1 (Th1)-induced cell-mediated autoimmune diseases. In the present study, to determine how LT, PF and FasL are involved in the pathogenesis of PM, we used immunohistochemical staining (IHC), reverse transcription polymerase chain reaction (RT-PCR), and in situ hybridization (ISH) on muscle specimens from patients with PM, amyotrophic lateral sclerosis (ALS), myotonic dystrophy (MyD) and controls (NC). There were many mononuclear cells (MNCs) immunoreactive for LT and some for PF and FasL within the fasciculus in PM muscles. On the other hand, only few or no LT-, PF- and FasL-positive cells were detected in MyD, ALS and NC muscles. The results of mRNA expression of these three molecules with RT-PCR were consistent with those using IHC methods. The number of MNCs positive for LT with ISH was far higher in PM compared to MyD, ALS and NC (P < 0.05 or 0.01). The MNCs located in the connective tissue or in the vicinity of necrotizing or non-necrotizing muscles were mainly LT mRNA and CD4 positive, while MNCs invading the non-necrotic fibers were mainly LT mRNA and CD8 positive. Our results indicated that the expression of LT was up-regulated in PM, and LT plays an important role in muscle injury and orchestrating the inflammatory reaction in PM. PMID- 11045676 TI - TUNEL-positive staining of surface contusions after fatal head injury in man. AB - In frontal lobe contusions obtained post mortem from 18 patients who survived between 6 h and 10 days after head injury, DNA fragmentation associated with either apoptotic and/or necrotic cell death was identified by the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nick end labelling (TUNEL) histochemical technique. Additional histological techniques were also used to identify regional and temporal patterns of tissue damage. TUNEL positive cells were present in both the grey and white matter of the contusion, where they peaked in number between 25 and 48 h, and were still identifiable at 10 days post injury. Fewer TUNEL-positive cells were observed in grey than in white matter; and most TUNEL-positive neurons in the grey matter demonstrated the morphological features of necrosis. However, the morphology of some TUNEL-stained neurons, and of TUNEL-stained oligodendroglia and macrophages in white matter was suggestive of apoptosis. Apoptosis was not seen in age- and sex-matched controls, none of whom had died from intracranial pathology or had pre-existing neurological disease. These findings suggest that multiple cell types in frontal lobe contusions exhibit DNA fragmentation and that both necrosis and apoptosis are likely to contribute to post-traumatic pathology. These findings provide further evidence that the observations made in animal models of traumatic brain injury have fidelity with clinical head injury. PMID- 11045675 TI - Major histocompatibility complex class II expression by activated microglia caudal to lesions of descending tracts in the human spinal cord is not associated with a T cell response. AB - Lesion-induced microglial/macrophage responses were investigated in post-mortem human spinal cord tissue of 20 patients who had died at a range of survival times after spinal trauma or brain infarction. Caudal to the spinal cord injury or brain infarction, a strong increase in the number of activated microglial cells was observed within the denervated intermediate grey matter and ventral horn of patients who died shortly after the insult (4-14 days). These cells were positive for the leucocyte common antigen (LCA) and for the major histocompatibility complex class II antigen (MHC II), with only a small proportion staining for the CD68 antigen. After longer survival times (1-4 months), MHC II-immunoreactivity (MHC II-IR) was clearly reduced in the grey matter but abundant in the white matter, specifically within the degenerating corticospinal tract, co-localising with CD68. In this fibre tract, elevated MHC II-IR and CD68-IR were still detectable 1 year after trauma or stroke. It is likely that the subsequent expression of CD68 on MHC II-positive microglia reflects the conversion to a macrophage phenotype, when cells are phagocytosing degenerating presynaptic terminals in grey matter target regions at early survival times and removing axonal and myelin debris in descending tracts at later survival times. No T or B cell invasion or involvement of co-stimulatory B7 molecules (CD80 and CD86) was observed. It is possible that the up-regulation of MHC II on microglia that lack the expression of B7 molecules may be responsible for the prevention of a T cell response, thus protecting the spinal cord from secondary tissue damage. PMID- 11045677 TI - Systemic hypothermia following spinal cord compression injury in the rat: an immunohistochemical study on MAP 2 with special reference to dendrite changes. AB - Systemic hypothermia has been shown to exert neuroprotective effects in experimental ischemic CNS models caused by vascular occlusions. The present study addresses the question as to whether systemic hypothermia has similar neuroprotective qualities following severe spinal cord compression trauma using microtubule-associated protein 2 (MAP2) immunohistochemistry combined with the avidin-biotin-peroxidase complex method as marker to identify neuronal and dendritic lesions. Fifteen rats were randomized into three equally sized groups. One group sustained thoracic laminectomy, the others severe spinal cord compression trauma of the T8-9 segment. The control group contained laminectomized animals submitted to a hypothermic procedure in which the esophageal temperature was reduced from 38 degrees C to 30 degrees C. The two trauma groups were either submitted to the same hypothermic procedure or kept normothermic during the corresponding time. All animals were sacrificed 24 h following the surgical procedure. The MAP2 immunostaining in the normothermic trauma group indicated marked reductions in MAP2 antigen in the cranial and caudal peri-injury zones (T7 and T10, respectively). This reduction was much less pronounced in the hypothermic trauma group. In fact, the MAP2 antigen was present in almost equally sized areas in both the hypothermic groups independent of previous laminectomy alone or the addition of trauma. Our study thus indicates that hypothermia has a neuroprotective effect on dendrites of rat spinal cords subjected to compression trauma. PMID- 11045678 TI - Global hypoxia per se is an unusual cause of axonal injury. AB - Irreversible hypoxic brain damage and axonal injury are present in over 90% of fatal blunt head injuries. Given the frequency of each, difficulties arise as to whether or not they are due to different mechanisms and, as such, can be separately recognised and quantified. Recent literature has raised the possible role of hypoxia in the formation of axonal bulbs. The present study of 17 cases of cardio-respiratory arrest, 12 of status epilepticus, 3 of carbon monoxide poisoning and 12 controls was designed to test the relationship between hypoxia and axonal injury and to test the hypothesis whether or not the two entities can be separated into primary and secondary forms of traumatic brain injury. Axonal damage was seen in 9/17 and 7/12 of the cases with cardiac arrest and status epilepticus, respectively, in most of whom there was also evidence of raised intracranial pressure (ICP). All 3 cases of carbon monoxide poisoning had evidence of white matter damage in keeping with the classical pattern of selective vulnerability. It is concluded that the great majority of axonal damage identified in cases dying after cardiac arrest and status epilepticus can be attributed to raised ICP and the vascular complications of internal herniation. However, in some cases, axonal damage was seen in the absence of an elevated ICP, although its amount and distribution were different from diffuse axonal injury. In many cases there was an increase in expression of neuronal beta amyloid precursor protein. PMID- 11045679 TI - Morphological changes following experimental intraventricular haemorrhage and intraventricular fibrinolytic treatment with recombinant tissue plasminogen activator. AB - Intraventricular haemorrhage (IVH) occurs in up to 50% of patients with primary intracerebral haemorrhage and aneurysmal subarachnoid haemorrhage. It is a significant and independent contributor to mortality and morbidity in these intracranial haemorrhages. Using a model of isolated IVH, we assessed the morphological changes induced by intraventricular bleeding and investigated the effects of intraventricular fibrinolytic treatment following IVH. IVH was induced in 32 pigs by intraventricular infusion of 10 ml autologous blood along with thrombin. The treatment group received an intraventricular injection of 1.5 mg (1 mg/ml) tissue plasminogen activator (tPA) following the injection of blood. The placebo group received the same volume of normal saline. Morphological examinations of the brains were carried out 7 days and 6 weeks following IVH. The ventricles were incompletely filled with blood and significantly enlarged in the placebo group 7 days after the IVH. In contrast, no residual intraventricular clots were visible in the animals treated with tPA, and the diameters of the lateral ventricles had returned to normal within 7 days. Marked losses of the ependymal covering of the ventricular walls were found in the placebo-treated animals, while the ependymal layer was largely intact in the animals treated with tPA. No haemorrhages induced by tPA were observed. The results indicate that intraventricularly administered tPA significantly enhances the lysis of intraventricular blood clots, accelerates the resolution of acute posthaemorrhagic hydrocephalus, and preserves the integrity of the ependymal layer. PMID- 11045680 TI - Alpha-synuclein accumulation in a case of neurodegeneration with brain iron accumulation type 1 (NBIA-1, formerly Hallervorden-Spatz syndrome) with widespread cortical and brainstem-type Lewy bodies. AB - We studied a 27-year-old woman who died after a 6-year history of progressive dementia, dystonia, ataxia, apraxia, spasticity, choreoathetosis, visual and auditory hallucinations, and optic atrophy. Magnetic resonance imaging showed decreased intensity in the globus pallidus, substantia nigra, and dentate nuclei in T2-weighted images, supporting the clinical diagnosis of neurodegeneration with brain iron accumulation type 1 (NBIA-1; formerly known as Hallervorden-Spatz syndrome). At autopsy the brain showed mild frontotemporal atrophy and discoloration of the globus pallidus and the substantia nigra pars reticularis. Histologically, features typical of NBIA-1 were found including widespread axonal spheroids and large deposits of iron pigment in the discolored regions. Additionally, excessive numbers of Lewy bodies (LBs) were found throughout all examined brain stem and cortical regions. LBs of both types, as well as Lewy neurites in this case of NBIA-1, were strongly labeled by antibodies against alpha-synuclein. These findings give further evidence that accumulation of alpha synuclein is generally associated with LB formation, i.e., in Parkinson's disease, dementia with Lewy bodies and NBIA-1. The case presented here is particularly notable for its high number of LBs in all areas of the cerebral cortex. PMID- 11045681 TI - Spinal cord glioneuronal tumor with "rosetted" neuropil islands and meningeal dissemination: a case report. AB - Distinctive glioneuronal tumors arising within the cerebrum and displaying neuropil-like islands and tumor cells immunoreactive for neuronal and glial antigens have recently been described. We report a similar tumor in the cervico thoracic region of the spinal cord in a 44-year-old woman that recurred 1 year later with dissemination to the lumbar dura and cauda equina. The tumor was composed of "rosetted" neuropil islands displaying immunoreactivity for synaptophysin, whereas the intervening tumor cells were more fibrillar and immunoreactive for GFAP. The tumor cell nuclei immediately surrounding these neuropil islands were immunoreactive to the newly characterized neuronal marker, anti-Hu. While several cases of neurocytomas have been described in the spinal cord, to the best of our knowledge, this is the first example of a glioneuronal tumor with "rosetted" neuropil islands to be reported in the spinal cord. PMID- 11045682 TI - Abnormal neuronal and glial argyrophilic fibrillary structures in the brain of an aged albino cynomolgus monkey (Macaca fascicularis). AB - An aged albino male cynomolgus monkey (Macaca fascicularis) more than 35 years old died after showing neurological signs including gait disturbance, trembling, drowsing tendency and a decrease in activity. Neuropathological examination revealed glial fibrillary tangles (GFTs) mainly distributed in the putamen, caudate nucleus, thalamic nuclei, substantia nigra, red nucleus, globus pallidus, trapezoid body, pyramid, pons and medulla oblongata of the brain, and neurofibrillary tangles (NFTs) in the thalamic nuclei. These structures were positively stained by the modified Gallyas-Braak (GB) method and immunostained for tau. The tau-positive argyrophilic GFTs were morphologically classified into four types, as in human cases, i.e., tufts of abnormal fibers (TAFs), thornshaped astrocytes (TSAs), glial coiled bodies (GCBs) and argyrophilic threads (ATs) depending on their GB profiles, and GCBs were the major structures in this case. Some of these structures were also immunoreactive for alpha-synuclein. The glial cells possessing the structures were negative for glial fibrillary acidic protein, a marker for astrocytes, indicating that the argyrophilic GFTs were present in oligodendroglia. In addition, marked neuronal loss and ubiquitin positive spheroid bodies were observed in the substantia nigra and globus pallidus. According to the characteristic distribution of the argyrophilic structures in neurons and glial cells as well as clinical signs, the monkey might have suffered from a neurodegenerative disease such as progressive supranuclear palsy (PSP). This is the first report of the occurrence of a neurodegenerative disease in a nonhuman animal. PMID- 11045683 TI - Evaluation of coronary artery bypass graft patency using three-dimensional reconstruction and flow study on electron beam tomography. AB - OBJECTIVES: To establish and evaluate two protocols for the noninvasive visualization and assessment of coronary artery bypass graft (CABG) patency on electron beam tomography (EBT). METHODS: Two hundred fourteen consecutive patients who underwent CABG surgery were scanned using both EBT angiography with three-dimensional reconstruction and EBT flow study with time-density curve analysis. RESULTS: There was a total of 589 CABGs evaluated in this study (10 grafts were excluded because of artifacts); 133 (98.5%) of 135 arterial grafts were patent, 345 (77.7%) of 444 saphenous vein grafts were patent. Within 5 years or 5-10 years after surgery, arterial graft patency exceeded venous graft patency (p<0.001). Three-dimensional EBT angiography achieved higher sensitivity, specificity, and accuracy (97.7%, 94.1%, and 96.7%, respectively) than EBT flow study (88.4%, 82.4%, and 85.2%, respectively) for evaluating occlusion or patency of CABGs. The intragraft flow of patent arterial and venous grafts were 4.9+/-2.2 ml/min/g and 6.9+/-2.8 ml/min/g, respectively (p<0.001). CONCLUSION: The combination of EBT three-dimensional reconstruction and flow study can be more effectively performed in the assessment of CABG anatomy and quantification of patent CABG flow. PMID- 11045685 TI - MRI of the pulmonary veins: comparison between 3D MR angiography and T1-weighted spin echo. AB - OBJECTIVE: The purpose was to determine the ability of three-dimensional (3D) magnetic resonance (MR) angiography to depict normal pulmonary veins in comparison with spin-echo MR imaging. MATERIALS AND METHODS: MR imaging of 40 patients with cardiovascular disease were reviewed. Patients with known pulmonary venous abnormalities were excluded. Using a standard GE 1.5-T magnet, axial T1 weighted spin-echo 5-mm-thick contiguous slices and 3D MR angiography (contiguous slice thickness of 2.5-3.5 mm, 20-30 c.c. of gadolinium bolus at 1-1.5 c.c./sec, 32-43-second breath-hold, coronal and sagittal plane acquisition) were evaluated retrospectively on separate occasions by two experienced radiologists. Multiplanar imaging projection was used for the identification of pulmonary veins. Each lung was considered to have two drainage veins: a superior vein and an inferior vein. Identification of a pulmonary vein was made by visualizing a connection with the left atrium. RESULTS: 143 pulmonary veins (87.5% +/-5.2) were identified at the level of the left atrium on T1-weighted spin-echo images, and 157 (98.1% +/-1.9) were identified on 3D MR angiography (p<0.01). Overall we identified by T -weighted spin-echo imaging 36 right upper, 38 right lower, 27 left upper, and 38 left lower pulmonary veins. By 3D MR angiography, we identified 38 right upper, 40 right lower, 39 left upper, and 40 left lower pulmonary veins. All four pulmonary veins were detected in 22 patients on spin echo imaging (55%) and in 37 patients (92.5%) on 3D MR angiography (chi = 3.81, p<0.05). CONCLUSION: A significant difference is demonstrated between 3D MR angiography and spin-echo MR imaging in identifying normal pulmonary veins. MR angiography provides a complete view of normal pulmonary venous anatomy and could be a valuable tool for the assessment of abnormal pulmonary venous drainage. PMID- 11045684 TI - Efficacy of MRI in complicated congenital heart disease with visceral heterotaxy syndrome. AB - PURPOSE: The authors' goal was to assess the diagnostic accuracy and clinical effect of MRI compared with echocardiography and catheterization in the evaluation of cardiac defects with situs ambiguous. METHOD: Twenty-two patients with visceral heterotaxy syndrome were included. RESULTS: Because situs determined by the relation between the pulmonary artery and bronchi showed most predominantly a tendency toward lateralization, this was regarded as the standard reference of situs determination. For the purpose of this study, patients were classified as having right isomerism (n = 13) or left isomerism groups (n = 9). MRI has several advantages compared with echocardiography or cardiac angiography for examining patients with situs ambiguous. (1) The bronchial, pulmonary arterial, and splenic situs can be readily determined, and discrepancies (n = 2) can be assessed easily. (2) Venoatrial connections are adequately imaged. In particular, all types of total and partial anomalous pulmonary venous return are delineated, regardless of whether restrictions of pulmonary blood flow or pulmonary venous obstructions are involved (n = 4). The courses of vertical veins were easily identified, and the prearterial position was revealed in only one of seven right isomerisms with total anomalous pulmonary venous return. The drain pattern of the hepatic vein can be visualized using three-dimensional spatial information and is useful for total cavopulmonary connection design. (3) Associated complicated cardiac anomalies, particularly the size or peripheral stenosis of the pulmonary arteries, may be evaluated, and this information is useful for palliative shunt operations. CONCLUSION: Because of its wide field of view and imaging, which is not restricted by associated anomalies, a thorough understanding of the cardiovascular anatomy of the situs ambiguous can be achieved using MRI, which is of considerable value in the surgical correction of this complicated anomaly. MRI can obviate or facilitate catheterization in these critically ill patients. PMID- 11045686 TI - Multislice cardiac spiral CT evaluation of atypical hypertrophic cardiomyopathy with a calcified left ventricular thrombus. AB - We report a case of a 43-year-old male patient with an atypical nonobstructive hypertrophic cardiomyopathy and a calcified left ventricular thrombus, and present results of multislice computed tomography (MSCT) using retrospective electrocardiograph gating, which is a new modality in cardiac imaging. Obtaining virtually motion-free images with a temporal resolution of 250 ms in an optimized heart scan MSCT allows functional imaging with evaluation of impaired systolic and diastolic left ventricular wall motion. PMID- 11045687 TI - Pulmonary tuberculosis: CT and pathologic correlation. AB - Typical CT findings of active postprimary pulmonary tuberculosis include centrilobular nodules and branching linear structures (tree-in-bud appearance), lobular consolidation, cavitation, and bronchial wall thickening. The CT findings of inactive pulmonary tuberculosis include calcified nodules or consolidation, irregular linear opacity, parenchymal bands, and pericicatricial emphysema. The typical appearance of primary tuberculosis on CT scans is homogeneous, dense, well-defined segmental or lobar consolidation with enlargement of lymph nodes in the hilum or the mediastinum. Miliary nodules may be seen in primary and postprimary tuberculosis. On CT, tuberculomas appear as a nodule with surrounding satellite nodules and internal cavitation on CT. Atypical radiologic manifestations of tuberculosis, encountered in as many as one third of the cases of adult-onset tuberculosis, are single or multiple nodules or masses, basilar infiltrates, miliary tuberculosis with diffuse bilateral areas of ground-glass opacity, and reversible multiple cysts. Underlying histopathologic findings of typical and atypical CT findings of tuberculosis are caseating granulomas or pneumonia in the active phase and fibrosis and dystrophic calcification in the inactive phase. PMID- 11045688 TI - Thoracic manifestations associated with advanced liver disease. AB - Advanced liver disease and portal hypertension may produce various intrathoracic complications that involve the pleural space, lung parenchyma, and pulmonary circulation. Dyspnea and arterial hypoxemia are the common clinical symptoms and signs in patients with such complications. In these patients, intrathoracic complications most often develop during the course of hepatic disease, but a few patients may be seen first with respiratory symptoms or radiographic abnormalities. Therefore, radiologists should be made aware of these disorders that occur in patients with chronic liver disease. In this article, the authors describe and illustrate the clinical and imaging spectrum of thoracic abnormalities associated with advanced liver disease and portal hypertension. PMID- 11045689 TI - Mediastinal lymphadenopathy in pulmonary fibrosis: correlation with disease severity. AB - OBJECTIVE: To evaluate the relationship between mediastinal lymph node enlargement and disease severity score in patients with pulmonary fibrosis. MATERIALS AND METHODS: A retrospective study included 30 patients with pulmonary fibrosis: idiopathic pulmonary fibrosis (n = 25), usual interstitial pneumonia (UIP) associated with collagen vascular disease (n = 4), and UIP associated with hepatitis C (n = 1). Disease severity was determined by a computed tomography (CT) scoring system. Each patient's lobe was scored by two radiologists on a scale of 0-5 for both ground glass opacity (GGO) and fibrosis. The presence, number, and sites of enlarged nodes (short axis > or = 10 mm) were assessed. CT severity scores were compared with total number of enlarged lymph nodes (L/Ns) and short axis diameter of the largest L/N (LLN). According to each severity score, patients were divided into two groups: the GGO-predominant group (n = 10) and the fibrosis-predominant group (n = 20). Total numbers of enlarged L/Ns and short axis diameter of LLN were compared in each group. RESULTS: Enlarged mediastinal L/Ns were present in 86%. Total severity score, GGO score, and fibrosis score strongly correlated with total number of enlarged L/Ns (p<0.05). Total severity score and GGO score correlated well with short axis diameter of LLN; however, the fibrosis score did not correlate with the short axis diameter of LLN. In respect to total number of enlarged L/Ns, the difference between the GGO group and fibrosis group was not apparent. In respect to the short axis diameter of LLN, the GGO group LLN was larger in diameter than the fibrosis group LLN (p<0.05). CONCLUSION: The greater the severity score of pulmonary fibrosis, the larger the total number of enlarged L/Ns. Those patients with more GGO had larger lymph nodes. PMID- 11045690 TI - T1 lung cancer on CT: frequency of extrathoracic metastases. AB - PURPOSE: To determine the frequency of extrathoracic metastases in T1 non-small cell lung cancer. METHOD: Ninety patients with T1 lung cancer identified on CT were included. Extrathoracic metastases were evaluated at the time of initial diagnosis and during a 1-year follow-up study. The frequency of metastases was compared in terms of cell type (squamous or nonsquamous), size (<2 cm or >2 cm), and the initial CT findings of the tumor. RESULTS: Extrathoracic metastases were identified in 12 (13%) of 90 patients at the time of diagnosis and in 10 patients at the 1-year follow-up study (total, 22 of 90 [24%] patients). Tumors with ground-glass opacity on CT were associated with a significantly lower prevalence of metastases (p = 0.042). The area of ground-glass opacity was seen in 1 of 13 (85%) patients with bronchioloalveolar carcinoma and in 12 of 53 (23%) patients with adenocarcinoma other than bronchioloalveolar carcinoma (p < 0.001). There was no significant difference in the prevalence of metastases between squamous and nonsquamous cell carcinoma, between tumors smaller than 2 cm (n = 17) and larger than 2 cm in diameter (n = 73) and between tumors with or without mediastinal nodal metastases (p>0.05). CONCLUSION: Extrathoracic metastases were apparent at the initial examination in 13% of patients and at the 1-year follow up examination in 11% of patients. The prevalence is significantly lower in tumors with ground-glass opacity. PMID- 11045691 TI - Pulmonary epithelioid hemangioendothelioma: atypical radiologic findings of a rare tumor with pathologic correlation. PMID- 11045692 TI - Mediastinal hemangioendothelioma: radiologic--pathologic correlation. PMID- 11045693 TI - MR dynamic enhancement of breast lesions: high temporal resolution during the first-minute versus eight-minute study. AB - PURPOSE: To investigate the value of the early phase of MR enhancement of breast lesions. METHOD: To study 63 breast lesions (size 5-45 mm in diameter) in 56 patients, whole-breast and lesion-targeted precontrast T1 -weighted gradient-echo 2D sequences were acquired. After intravenous injection of Gd-DTPA (0.1 mmol/Kg), four targeted scans, each every 15 seconds during the first minute (1-m), and seven whole-breast scans, each every minute up to 8 minutes (8-m), were performed. The subtraction technique was used, and percent enhancement curves were obtained. The final diagnosis was obtained by histology for 36 lesions, including 28 malignancies, and by fine-needle aspiration cytology and at least 1 year negative follow-up for the remaining 27 benign lesions. RESULTS: Significant differences in enhancement between malignant and benign lesions were found using both techniques (p<0.0001). However the ratio between the median enhancement of malignant lesions and that of benign lesions was 6.7 (15 s), 4.8 (30 s), 4.6 (45 s), and 3.8 (60 s), descending from 4.3 to 2.5 from the second to the eighth minute. The overlap between the malignant and benign curves was 9% of the malignant range with the 1-m technique, and 50% with the 8-m technique. Three blinded observers obtained a 100% sensitivity with both techniques and a specificity of 94-97% with the 1-m technique and 83-89% with the 8-m technique. CONCLUSION: The first minute of Gd-enhancement allows a more prominent differentiation between malignant and benign breast lesions than the following times. PMID- 11045694 TI - Functional MRI of language-related activation in left frontal schizencephaly. AB - Brain activation associated with covert verb generation was studied in a right handed patient with an asymptomatic left frontal schizencephaly by functional magnetic resonance imaging. Activation of the dysplastic neuronal tissue lining the cleft and the adjacent cortex was found, indicating participation of the malformed region in physiologic cerebral functions. This finding may have clinical impact in patients with medically intractable seizures due to malformations of the cortical development prior to epilepsy surgery. PMID- 11045695 TI - Heroin-induced spongiform leukoencephalopathy: value of diffusion MR imaging. AB - The diffusion-weighted (DW) magnetic resonance (MR) imaging findings of a patient with subacute stage of heroin-induced vacuolating myelinopathy are reported. The diffuse decrease of apparent diffusion coefficient (ADC) of the white matter on DW imaging is attributed to restricted water diffusion, which is known to be caused by fluid entrapment within the myelin lamellae without demyelination. PMID- 11045696 TI - Rotator cuff interval: evaluation with MR imaging and MR arthrography of the shoulder in 32 cadavers. AB - PURPOSE: The purpose of this work was to establish the optimal means of evaluation of the rotator cuff interval (RCI) and rotator interval capsule and demonstrate normal anatomy of the RCI using MR imaging and MR arthrography. METHOD: MR arthrography was performed in 32 cadaveric shoulders. In 20 cases, MR imaging was completed prior to arthrography. Pre- and postarthrography studies included standard imaging planes. Images were evaluated by the consensus of two musculoskeletal radiologists with attention to the RCI, rotator interval capsule (measurements on postarthrographic studies), and crossing structures. In five cases, specialized imaging planes were performed after arthrography. RESULTS: The RCI, rotator interval capsule, and crossing structures were best evaluated by MR arthrography. The anteroposterior dimension of the rotator interval capsule could be best depicted on postarthrogram images. CONCLUSION: MR arthrography, with both standard and specialized imaging planes, is a useful way to evaluate the RCI, the rotator interval capsule, and its crossing structures. PMID- 11045697 TI - Osteochondritis dissecans of the tarsal navicular bone: imaging findings in four patients. AB - We report the imaging characteristics of osteochondritis dissecans of the tarsal navicular bone in four cases and review the current literature. Its radiological findings are similar to osteochondritis dissecans found in other sites: focal lucency that disrupts the sharp cortical line, the presence of sclerosis, and cortical depression. PMID- 11045698 TI - Helical CT findings in patients with hepatocellular carcinoma treated with percutaneous ablation procedures. AB - Nonsurgical treatment of hepatocellular carcinoma is used worldwide as a result of the early detection and slow growth of this tumor in patients with chronic liver disease. Multiple-phase helical computed tomography is a commonly used method for evaluating the main features related to percutaneous ablation procedures: nodular changes, tumor necrosis, parenchymal changes, complications, and tumor recurrence. Knowledge of all features recognizable after local ablation therapy is mandatory to avoid diagnostic pitfalls and to optimally assess treatment response. PMID- 11045699 TI - Septic thrombophlebitis of the mesenteric and portal veins: CT imaging. AB - Pylephlebitis or septic thrombophlebitis of the portal vein and its tributaries is an acute ascending infection arising often from a primary gastrointestinal inflammatory lesion. Common primary sources of infection are diverticulitis, appendicitis, and infected pancreatic necrosis. CT imaging can diagnose this complication at an early stage and can significantly improve the previously reported high mortality and morbidity rates associated with this condition. PMID- 11045700 TI - Low attenuation intratumoral matrix: CT and pathologic correlation. AB - Diffuse low attenuation lesions on CT scans are caused by various pathologic conditions, especially differences of the intratumoral matrix. Low attenuation on CT scans was more defined than fat density and less than muscle density, the so called near water density. We present representative cases of low attenuation lesions in correlation with their pathologic features. Intratumoral matrix showing diffuse low attenuation on CT was classified into eight groups: fatty degeneration/fatty metamorphosis, intracellular high lipid content, large amounts of lipid-laden macrophages, mucin-producing tumor, myxoid degeneration/myxoid matriX, massive necrosis, true cystic growth of the neoplasm, and massive edema. Lesions with fatty degeneration or intracellular high lipid content could be shown as negative CT attenuation. Knowledge of the neoplasms or disease processes that tend to have such pathology will lead to correct interpretation of CT scans of various organs. PMID- 11045701 TI - Siderotic nodules at MR imaging: regenerative or dysplastic? AB - OBJECTIVE: To determine if iron containing "siderotic" nodules detected at magnetic resonance (MR) imaging are regenerative (RN) or dysplastic (DN) and to attempt to identify features that may distinguish them. MATERIAL AND METHODS: MR imaging (1.5 T) was performed on 77 cirrhotic patients who underwent orthotopic liver transplantation within 0-117 days (mean 30 days) of MR imaging. Two readers retrospectively evaluated breath-hold gradient-echo pulse sequences (echo time > or =9.0 ms, flip angle < or =45 degrees) for the presence of hypointense nodules, which were classified as micronodular (< or =3 mm), macronodular (>3 mm), or mixed. Nodule distribution was classified as focal (<5), scattered (5-20), or diffuse (>20) per slice. Thin section pathologic correlation was available in all cases, and Prussian blue iron stains were performed. RESULTS: Of 35 patients with pathologically proven siderotic nodules, 10 (29%) had at least 2 siderotic DN. MR detected siderotic nodules in 10 of 10 (100%) patients with siderotic DN and RN, and in 18 of 25 patients (72%) with siderotic RN only. CONCLUSION: Siderotic RN cannot be reliably distinguished from siderotic DN with MR imaging, and therefore the widely used term "siderotic regenerative nodule" should be avoided and replaced by "siderotic nodule." PMID- 11045702 TI - Two-phase helical CT for detection of early gastric carcinoma: importance of the mucosal phase for analysis of the abnormal mucosal layer. AB - PURPOSE: To evaluate the efficacy of two-phase dynamic helical computed tomography (CT), including the gastric mucosal phase, for detection of early gastric carcinoma with typical hyperattenuating and atypical nonhyperattenuating enhancement patterns. METHOD: Two-phase helical CT scanning was performed using the water-filling method as negative oral contrast material for 212 patients with proven adenocarcinoma on endoscopic biopsy. Two gastrointestinal radiologists prospectively analyzed the focal alterations of the inner hyperattenuating mucosal layer and the outer hypoattenuating layer before the information obtained at barium study and pathologic examination was available. The first, so-called mucosal phase was obtained at 38-45 seconds after the start of intravenous injection of contrast material for a total of 150 ml/sec at a rate of 4 ml/sec to obtain maximum enhancement of the inner mucosal layer. The second delayed phase was obtained at 3 minutes. RESULTS: Fifty-four cases of early gastric cancer were suspected on two-phase helical CT preoperatively. Postoperatively, 49 cases of early gastric cancer were pathologically confirmed. The detection rate for the typical hyperattenuating early gastric cancer, that is the type I enhancement pattern defined as the localized thickening of the inner hyperattenuating layer, using two-phase helical CT was 18% (9/49 patients). The type 2 enhancement pattern, defined as the focal interruption of the inner hyperattenuating mucosal layer without abnormal enhancement of the outer hypoattenuating layer on the mucosal phase, was seen in 15 cases. These were pathologically confirmed as early gastric cancer IIb + IIc (three patients), IIc (four patients), IIc + IIa (one patient), IIc + III (three patients), IIb + IIc (one patient), and advanced cancer (T2) lesions (three patients). The type 3 enhancement pattern was defined as the focal polypoid protrusion of the inner hyperattenuating layer without abnormal enhancement of the outer thickened hypoattenuating layer on the mucosal phase, and was seen in seven patients who were pathologically confirmed with early gastric cancer IIb + IIc (three patients), IIc + IIa (one patient), and IIc + lIb (three patients). The lesions became less distinct on the delayed phase. Five T2 advanced gastric cancers were falsely interpreted as early gastric cancer. The detection rate for early gastric cancer after considering type 2 and 3 atypical enhancement patterns was increased to 57% (28/49 patients). CONCLUSION: Helical CT with two-phase scan including the mucosal phase was efficient for identifying the enhancement patterns of early gastric cancer. PMID- 11045703 TI - Abdominal lymphoma staging: is MR imaging with T2-weighted turbo-spin-echo sequence a diagnostic alternative to contrast-enhanced spiral CT? AB - PURPOSE: To evaluate the diagnostic value of MR imaging in abdominal lymphoma staging, in comparison with that of the established spiral CT, within the context of a prospective study. METHODS: 50 patients with non-Hodgkin (n = 27) and Hodgkin lymphoma (n = 23) were examined with a plain T2-weighted TSE sequence (parameters: TE 90 ms, TR >2.500 ms, slice thickness 8 mm, slice interval 0.8 mm, ETL 20, NEX 4), and with spiral CT following oral and intravenous administration of contrast agent. RESULTS: Both CT and MR imaging agreed in showing abdominal lymphomas in 34/50 cases. The size of the detected lymphomas was between 1.5-9 cm (mean: 4.3 +/-2.2 cm). In the analysis of the individual lymph node sites, CT showed involvement of the paraaortic lymph nodes in 29/50 patients, compared with 28/50 in MRI, and involvement of the portal lymph nodes in 15/50, compared with 12/50. Both techniques showed the iliac lymph nodes in 21/50 patients, the inguinal lymph nodes in 10/50, and the mesenteric lymph nodes in 11/50. Both techniques also showed focal organ lesions in 12/50 cases. CONCLUSIONS: In the staging of abdominal lymphomas, MR imaging with a T2-weighted TSE sequence can be regarded as equal to spiral CT in the detection of lymph adenopathy and the demonstration of focal organ lesions. In addition to the absence of ionizing radiation, the advantage of MR imaging is that there is no necessity for oral or intravenous administration of contrast agent. PMID- 11045704 TI - Assessment of the presence and severity of esophagogastric varices by splenic index in patients with liver cirrhosis. AB - PURPOSE: To determine whether spleen size is related to the severity of esophageal varices or associated gastric varices and liver functions in patients with cirrhosis. METHOD: The authors retrospectively studied spleen size on CT (splenic index [SI] = length x width x height of the spleen), liver functions, and the results of esophagogastric endoscopy in 110 patients with cirrhosis. They also analyzed SI in 112 controls. RESULTS: In controls, body weight, height, and age affected the SI. The SI in patients with uncompensated cirrhosis was greater compared with the SI in those with well-compensated disease (p = 0.0363). The SI in patients with esophageal varices was greater than in patients without esophageal varices (p<0.0001), but patients with and without gastric varices had similar SI values. The SI in patients with the red color signs (red wale marking, cherry red spot, and hematocystic spot) on esophageal varices or with risky varices (enlarged tortuous varices with beady, nodular, or tumor shape associated with red color signs) was greater than in patients without these signs (p = 0.0029 and p = 0.0030, respectively). CONCLUSION: The SI is a good indicator of the severity of esophageal varices and hepatic functional reserve in patients with cirrhosis. PMID- 11045705 TI - Unenhanced helical CT using increased pitch for suspected renal colic: an effective technique for radiation dose reduction? AB - PURPOSE: To determine the accuracy and utility of unenhanced helical CT for suspected renal colic, using a pitch of either 2.5 or 3.0. METHODS: 59 consecutive patients underwent unenhanced helical CT. 5 mm contiguous images were obtained at a kVP of 120 and an mA of 260. Thirty-four patients were imaged at a pitch of 2.5, and 25 patients were imaged at a pitch of 3.0. Two radiologists, an attending (reader 1), and a second-year resident (reader 2), independently and retrospectively reviewed the CT images, blinded to the clinical outcome. The presence or absence of a ureteral stone was recorded and image quality was graded. A third radiologist determined accuracy for each reader. Average entrance exposure was estimated using a CT phantom at a variety of pitches. RESULTS: Overall sensitivity, specificity, and accuracy for reader 1 were 91, 96, and 93%. For reader 2, they were 86, 93, and 90%. There was no significant difference in accuracy using a pitch of 3.0 compared with 2.5 for either reader. Readers 1 and 2 rated image quality at 2.5 pitch as excellent for 88 and 76% of scans, respectively; at 3.0 pitch the scans were rated by both readers as excellent for 40% and acceptable for 60%. Average entrance exposures were estimated at 461, 553. and 913 mR at pitches of 3.0, 2.5, and 1.5. CONCLUSION: Increasing the pitch on unenhanced helical CT for suspected renal colic to 2.5 or 3.0 appears to be an effective method of reducing radiation dose. Although accuracy of the technique did not significantly change using a pitch of 3.0 in one group of patients, compared with a pitch of 2.5 in another group of patients, image quality did decrease. PMID- 11045706 TI - Intrahepatic portal venous variations: demonstration by helical CT during arterial portography. AB - PURPOSE: We assessed the prevalence and types of intrahepatic portal venous variations by helical computed tomography performed with arterial portography (CTAP). METHODS: In 192 patients without evidence of vascular invasion or distortion, CTAP images were reviewed retrospectively to identify portal venous variations. RESULTS: Of the 192 patients examined, 10 (5.2%) had trifurcation, 5 (2.6%) had a right posterior segmental branch arising from the main portal vein, 5 (2.6%) had an absence of the horizontal segment of the left portal vein, and 1 (0.5%) had an absence of the left lateral segmental portal branch. Of the patients without a horizontal segment, two had a right-sided ligamentum teres associated with malposition of the gallbladder, while another had complete ramification of intrahepatic portal branches from an umbilical vein-like segment. In the patient missing the left lateral segmental branches, the right portal vein segments were subcapsularly located. CONCLUSION: Variations of the intrahepatic portal veins can be recognized on CTAP imaging. tomography-Portal vein, computed tomography. PMID- 11045707 TI - Cholangiolocellular carcinoma of the liver: CT and MR findings. AB - The authors report two cases of surgically proved cholangiolocellular carcinoma of the liver. Marked contrast enhancement was observed at the periphery of the tumor on CTs and MRIs obtained during the hepatic arterial and portal venous phases, with concentric filling on the delayed images. On T1-weighted and T2 weighted MRIs, the tumor was, respectively, hypointense and hyperintense, with a central hypointense area. Therefore, helical CT and MRI features of these cholangiolocellular carcinomas were thought to be similar to those of cholangiocarcinoma. PMID- 11045708 TI - The costs of CT procedures in an academic radiology department determined by an activity-based costing (ABC) method. AB - PURPOSE: The purpose of this work was to determine the costs of computed tomography (CT) procedures in a large academic radiology department, including both professional (PC) and technical (TC) components, by analyzing actual resource consumption using an activity-based costing (ABC) method and comparing them with Medicare payments. METHOD: Over a 12 month period from July 1, 1996, to June 30, 1997, 1,011 CT procedures, representing 16 Physicians' Current Procedural Terminology (CPT) codes and 98.3% of CT studies performed, were carefully observed by a research assistant trained in ABC methodology. Information collected during these time and motion studies included personnel/machine time and direct materials used. Actual resource units used during the different activities in each CT procedure were valued using appropriate cost drivers. Unit values for both direct and overhead costs were calculated: the cost of an individual procedure equaled the sum of component costs. Costs were compared with PC and TC payments according to the 1997 Medicare Fee Schedule. RESULTS: Total costs of CPT codes 70450 (CT Head unenhanced), 71260 (CT Chest enhanced), and 74160 (CT Abdomen enhanced), which represented 71.2% of CT studies performed, were $189.19, $273.53, and $343.20, respectively. For all 16 nonmodified CPT codes analyzed, Medicare's professional reimbursement was less than the professional cost, whereas its technical reimbursement exceeded respective cost in 14 of the 16 codes. CONCLUSION: In the setting and time period studied, Medicare underreimbursed professional costs while overreimbursing technical costs. PMID- 11045709 TI - The nomenclature and sectional imaging anatomy III: capsular membranes and minor spinal ligaments. AB - This paper is the third in a series of three that organizes the complex anatomy of the cervical, thoracic, and lumbar spinal ligaments. It describes and color codes the anatomy and nomenclature of the capsular membranes and minor spinal ligaments. The first two articles describe the dorsal and ventral ligaments, respectively. PMID- 11045710 TI - On spectral relaxation in proteins. AB - During the past several years there has been debate about the origins of nonexponential intensity decays of intrinsic tryptophan (trp) fluorescence of proteins, especially for single tryptophan proteins (STP). In this review we summarize the data from diverse sources suggesting that time-dependent spectral relaxation is a ubiquitous feature of protein fluorescence. For most proteins, the observations from numerous laboratories have shown that for trp residues in proteins (1) the mean decay times increase with increasing observation wavelength; (2) decay associated spectra generally show longer decay times for the longer wavelength components; and (3) collisional quenching of proteins usually results in emission spectral shifts to shorter wavelengths. Additional evidence for spectral relaxation comes from the time-resolved emission spectra that usually shows time-dependent shifts to longer wavelengths. These overall observations are consistent with spectral relaxation in proteins occurring on a subnanosecond timescale. These results suggest that spectral relaxation is a significant if not dominant source of nonexponential decay in STP, and should be considered in any interpretation of nonexponential decay of intrinsic protein fluorescence. PMID- 11045711 TI - Rotational relaxation of neutral red in alkanes: effect of solvent size on probe rotation. AB - Rotational reorientation times of a polar molecule neutral red (NR) have been measured in n-alkanes using steady-state fluorescence depolarization technique. The rotational dynamics of NR in alkanes is described by the Stokes-Einstein Debye hydrodynamic theory with slip boundary condition. However, we have observed that as the size of the solvent molecule becomes bigger than the size of the solute molecule, the probe molecule experiences reduced friction and the experimentally measured reorientation times are shorter than those predicted by the hydrodynamic theory. These size effects have been analyzed using quasihydrodynamic theories. PMID- 11045712 TI - The location of tryptophan, N-acetyltryptophan and alpha-chymotrypsin in reverse micelles of AOT: a fluorescence study. AB - The spectroscopic properties of alpha-chymotrypsin (alpha-Chym), L-tryptophan (Trp) and N-acetyl-L-tryptophan (NAT) solubilized in hydrated reverse micelles of sodium bis(2-ethylhexyl) sulfosuccinate in iso-octane were followed by fluorescence as a function of the amount of intramicellar water and initial pH. The lack of pH dependence observed for Trp in these systems, as opposed to what occurs in bulk water, and the similarities found for the protein in both media foresee different locations of these probes. In reverse micelles, fluorescence quenching studies using acrylamide emphasize the existence of structural alterations within the protein when its global charge changes from positive (pH = 7) to negative (pH = 10). The ensemble of the data points to an interfacial location of the zwitterionic Trp, an intermediate region of less tightly bound water for the location of the anionic Trp and NAT and an almost bulk water environment for alpha-Chym. PMID- 11045713 TI - A laser flash photolysis and pulse radiolysis study of primary photochemical processes of flumequine. AB - The 355 nm laser flash photolysis of argon-saturated pH 8 phosphate buffer solutions of the fluoroquinolone antibiotic flumequine produces a transient triplet state with a maximum absorbance at 575 nm where the molar absorptivity is 14,000 M(-1) cm(-1). The quantum yield of triplet formation is 0.9. The transient triplet state is quenched by various Type-1 photodynamic substrates such as tryptophan (TrpH), tyrosine, N-acetylcysteine and 2-deoxyguanosine leading to the formation of the semireduced flumequine species. This semireduced form has been readily identified by pulse radiolysis of argon-saturated pH 8 buffered aqueous solutions by reaction of the hydrated electrons and the CO2*- radicals with flumequine. The absorption maximum of the transient semireduced species is found at 570 nm with a molar absorptivity of 2,500 M(-1) cm(-1). In argon-saturated buffered solutions, the semireduced flumequine species formed by the reaction of the flumequine triplet with TrpH stoichiometrically reduces ferricytochrome C (Cyt Fe3+) under steady state irradiation with ultraviolet-A light. In the presence of oxygen, O2*- is formed but the photoreduction of Cyt Fe3+ by O2*- competes with an oxidizing pathway which involves photo-oxidation products of TrpH. PMID- 11045714 TI - Alteration of the endocytotic pathway by photosensitization with fluoroquinolones. AB - The endocytotic pathway is profoundly altered by the UVA-induced photosensitization of HS 68 fibroblasts by the fluoroquinolone (FQ) antibiotics lomefloxacin, BAYy 3118, norfloxacin and ciprofloxacin, which preferentially localize in lysosomes. The endocytosis of low-density lipoproteins (LDL) loaded with two carbocyanine dyes compatible for effective Forster-type resonance energy transfer (FRET), namely 3,3'-dioctadecyloxacarbocyanine perchlorate (DiO) as the donor and 1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) as the acceptor, has been used as a model system. Binding of LDL to their cell surface receptors is impaired by irradiation with 10 J cm(-2) of UVA and/or treatment with 250 microM BAYy 3118 during 2 h. Perturbation of the plasma membrane by the FQ is revealed by the change in the rate of exchange of DiO from the LDL to the cell membrane as compared to untreated cells. The lysosomal degradation of LDL, demonstrated by the disappearance of FRET between DiO and DiI, is partly inhibited by the FQ. The actin filament network, involved in the fusion of mature endosomes with lysosomes, is readily destroyed upon photosensitization with the four FQ. However, actin depolymerization can be avoided by incubation of the cells with trans-epoxysuccinyl-1-leucylamido-(4 guanidino)butane, an inhibitor of lysosomal cathepsins prior to FQ photosensitization. All these data suggest that several components of the endocytotic pathway are impaired by photosensitization with these FQ. PMID- 11045715 TI - Photooxidation of cedrelone, a tetranortriterpenoid from Toona ciliata. AB - Cedrelone, a tetranortriterpenoid on photolysis by UV light yields a true photooxidation product 3 [14 beta,15beta,22beta,23beta-diepoxy-6-hydroxy-1,5, 20(22)-meliatriene-2,7,21-trione] whose structure is well established by NMR studies and confirmed by X-ray crystallography, along with product 4 [14 beta,15beta-epoxy-6,23-dihydroxy-1,5,20(22)-meliatriene-2,7, 21-trione]. Addition of rose bengal increases the rate of photooxidation whereas DABCO decreases rate of photolysis proving the involvement of singlet oxygen in the photooxygenation. Both the photoproducts exhibited antifeedant activity. PMID- 11045716 TI - Photochemical studies on xanthurenic acid . AB - The tryptophan metabolite xanthurenic acid (Xan) has been isolated from aged human cataractous lenses. The photophysical properties of Xan were examined to determine if it is a potential chromophore for age-related cataractogenesis. We found that Xan produces singlet oxygen (psi delta = 0.17 in CD3OD) with the same efficiency as the lenticular chromophore N-formyl kynurenine and quenches singlet oxygen at a rate similar (2.1 x 10(7); CD3OD) to other tryptophan metabolites found in the eye. As the mechanisms of induction of cataracts may also involve redox reactions, the interactions of hydrated electrons (e(aq)-), the azide radical (N3*) and hydroxyl radical (OH*) with Xan were studied using the technique of pulse radiolysis. The reaction rate constants of e(aq)-, N3* and OH* with Xan were found to be of the same order of magnitude as other tryptophan metabolites. The rate constant for reaction of Xan with e(aq)- solvated electrons was found to be diffusion controlled (k = 1.43 x 10(10) M(-1) s(-1); the reaction with N3* was very fast (k = 4.0 x 10(9) M(-1) s(-1)); and with OH* was also near diffusion controlled (k = 1.0 x 10(10) M(-1) s(-1)). Superoxide O2*- production by irradiated Xan in methanol was detected by electron paramagnetic resonance and substantiated by determining that the enhanced rate of oxygen consumption of Xan irradiated in the presence of furfuryl alcohol was lowered by superoxide dismutase. PMID- 11045717 TI - Photochemical and photobiological properties of new bispsoralen derivatives (Bis[PsCn]PIP, n = 4, 6, 8). AB - Bispsoralen derivatives possessing two psoralens and one piperazine molecule, 1,4 bis[n'-(8-psoralenoxy) alkyl] piperazine (Bis[PsCn]PIP, n = 4, 6, 8), show high water solubility, efficient intercalation into DNA and good photocrosslinking efficiency of DNA. Bis(PsC4)PIP shows high lethality on bacteriophage T7 and can effectively inhibit the amplification of DNA by stopping the polymerase chain reactions in a short period of irradiation time. PMID- 11045718 TI - Fluorometric analysis of DNA unwinding (FADU) as a method for detecting repair induced DNA strand breaks in UV-irradiated mammalian cells. AB - Fluorometric analysis of DNA unwinding (FADU assay) was originally designed to detect X-ray-induced DNA damage in repair-proficient and repair-deficient mammalian cell lines. The method was modified and applied to detect DNA strand breaks in Chinese hamster ovary (CHO) cells exposed to ionizing radiation as well as to UV light. Exposed cells were allowed to repair damaged DNA by incubation for up to 1 h after exposure under standard growth conditions in the presence and in the absence of the DNA synthesis inhibitor aphidicolin. Thereafter, cell lysates were mixed with 0.15 M sodium hydroxide, and DNA unwinding took place at pH 12.1 for 30 min at 20 degrees C. The amount of DNA remaining double-stranded after alkaline reaction was detected by binding to the Hoechst 33258 dye (bisbenzimide) and measuring the fluorescence. After exposure to X-rays DNA strand breaks were observed in all cell lines immediately after exposure with subsequent restitution of high molecular weight DNA during postexposure incubation. In contrast, after UV exposure delayed production of DNA strand break was observed only in cell lines proficient for nucleotide excision repair of DNA photoproducts. Here strand break production was enhanced when the polymerization step was inhibited by adding the repair inhibitor aphidicolin during repair incubation. These results demonstrate that the FADU approach is suitable to distinguish between different DNA lesions (strand breaks versus base alterations) preferentially induced by different environmental radiations (X-rays versus UV) and to distinguish between the different biochemical processes during damage repair (incision versus polymerization and ligation). PMID- 11045719 TI - UVAI-induced edema and pyrimidine dimers in murine skin. AB - The induction of edema and pyrimidine dimers in epidermal DNA was determined in the skin of SKH:HR1 mice exposed to graded doses of ultraviolet radiation AI (UVAI; 340-400 nm). Exposure to UVAI induced 1.6 +/- 0.08 x 10(-6) (mean +/- standard error of mean) pyrimidine dimers per 10(8) Da of DNA per J/m2. Edema in irradiated animals was determined as an increase in skinfold thickness. A dose of 1.8 x 10(6) J/m2 of UVAI that resulted in a 50% increase in skinfold thickness (SFT50%) would have induced 1.0 x 10(5) dimers per basal cell genome. A similar increase in SFT induced by full spectrum solar ultraviolet radiation (290-400 nm) would accompany the induction of 11.0 x 10(5) pyrimidine dimers per basal cell genome. These results support a hypothesis that UVAI-induced pathological changes of the skin are mediated through the formation of nondimer photoproducts. PMID- 11045720 TI - Annual and interannual behavior of solar ultraviolet irradiance revealed by broadband measurements. AB - This research examines the behavior of ground-level solar UV radiation as measured by eight broadband meters in the continental United States during the period from late 1994 to late 1998. The goal is to define the variability that occurs in UV irradiance over time scales ranging from one to several years. The monthly integrated irradiances, from latitude 32 degrees N to 47 degrees N, contain large annual cycles and latitudinal gradients which depend on season. Seven of the eight sites show a maximum in July, a behavior related to proximity to the summer solstice, with modifications associated with the annual cycle in column ozone. A large interannual variability in monthly integrated irradiance appears over the 4 year period studied. A comparison of corresponding months during different years shows differences in irradiance of 20% or more in one third of the cases analyzed. When the solar zenith angle (SZA) is held fixed in the range 60-65 degrees, a substantial annual cycle in UV irradiance remains where the maximum monthly mean irradiance is 1.4-1.9 times the minimum, depending on location. Furthermore, the annual cycle at fixed SZA is not in phase with the normal seasonal cycle. Maximum irradiances at fixed SZA tend to occur in the October to December period, while minima cluster in April through July. The annual cycle in ozone, with maximum column values in spring and minima in autumn, explains the general character of the fixed-SZA data, although changes in cloudiness are significant contributors to interannual variability. PMID- 11045721 TI - Exposure to ultraviolet radiation enhances mortality and pathology associated with influenza virus infection in mice. AB - Ultraviolet radiation (UVR) causes systemic immune suppression, decreasing the delayed type and contact hypersensitivity responses in animals and humans and enhancing certain mycobacterial, parasitic and viral infections in mice. This study tests the hypothesis that prior exposure to UVR enhances influenza infections in mice. BALB/c female mice were exposed to 0-8.2 kJ/m2 of UVR. Exposed and unexposed mice were infected intranasally three days later with 150 300 plaque-forming units/mouse (lethal dose (LD)20-LD40) of mouse-adapted Hong Kong Influenza A/68 (H3N2) virus or sham infected with 50 microL Hanks' balanced salt solution/mouse. Mortality from viral infection ranged from 25-50%. UVR exposure increased virus-associated mortality in a dose-dependent manner (up to a two-fold increase at 8.2 kJ/m2). The increased mortality was not associated with bacterial pneumonia. The highest dose of UVR also accelerated the body weight loss and increased the severity and incidence of thymic atrophy associated with influenza infection. However, UVR treatment had little effect on the increase in lung wet weight seen with viral infection, and, to our surprise, did not cause an increase in virus titers in the lung or dissemination of virus. The mice died 5-6 days after infection, too early for adaptive immune responses to have much impact. Also, UVR did not interfere with the development of protective immunity to influenza, as measured by reinfection with a lethal challenge of virus. Also, cells adoptively transferred from UVR or untreated mice were equally protective of recipient mice challenged with a lethal dose of virus. The mice resemble mice succumbing to endotoxin, and influenza infection increased the levels of tumor necrosis factor alpha (TNF-alpha) in bronchoalveolar lavage fluid and serum cortisol levels; however, UVR preexposure did not increase either of these responses to the virus. The results show that UVR increased the morbidity, mortality and pathogenesis of influenza virus in mice without affecting protective immunity to the virus, as measured by resistance to reinfection. The mechanism of enhanced mortality is uncertain, but the data raises concerns that UVR may exacerbate early responses that contribute to the pathogenesis of a primary viral infection. PMID- 11045722 TI - Degradation of the photosystem I complex during photoinhibition. AB - Exposure of isolated photosystem I (PSI) complexes to illumination (2300 microE m(-2) s(-1)) for various periods of time resulted in striking changes in their absorption spectra. A 6 nm blueshift of the absorption maximum in the red was detected after 100 min illumination. The fourth derivative of the absorption spectra verifies that the main change of the red peak was attributed to the 682 nm absorption band. Further, it was also shown that a shoulder in the absorption spectra located around 470 nm decreased after the first 5 min of illumination and almost disappeared after 40 min illumination, suggesting that chlorophyll b bound to light-harvesting complex I (LHCI) is also sensitive to excess light. A maximum inhibitory effect on the oxygen uptake rates and a strong stimulation were observed when the PSI complexes were exposed to illumination for about 20 and 40 min, respectively. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis shows that LHCI-680 started to degrade during the first 5 min of illumination and almost completely disappeared after 40 min of illumination. These observations demonstrated that LHCI was more sensitive to illumination than the PsaA/B subunits which also presented some degradation signs after 40 min illumination. In addition, insoluble-cohesive-denatured proteins also appeared between the stacking and resolving gel after prolonged illumination (100 min). A photoprotective function of LHCI for the PSI reaction center is proposed. PMID- 11045723 TI - Inhibition of photosynthetic electron transport by UV-A radiation targets the photosystem II complex. AB - We have studied the inhibition of photosynthetic electron transport by UV-A (320 400 nm) radiation in isolated spinach thylakoids. Measurements of Photosystem II (PSII) and Photosystem I activity by Clark-type oxygen electrode demonstrated that electron flow is impaired primarily in PSII. The site and mechanism of UV-A induced damage within PSII was assessed by flash-induced oxygen and thermoluminescence (TL) measurements. The flash pattern of oxygen evolution showed an increased amount of the S0 state in the dark, which indicate a direct effect of UV-A in the water-oxidizing complex. TL measurements revealed the UV-A induced loss of PSII centers in which charge recombination between the S2 state of the water oxidizing complex and the semireduced Q(A)- and Q(B)- quinone electron acceptors occur. Flash-induced oscillation of the B TL band, originating from the S2Q(B)- recombination, showed a decreased amplitude after the second flash relative to that after the first one, which is consistent with a decrease in the amount of Q(B)- relative to Q(B) in dark adapted samples. The efficiency of UV-A light in inhibiting PSII electron transport exceeds that of visible light 45-fold on the basis of equal energy and 60-fold on the basis of equal photon number, respectively. In conclusion, our data show that UV-A radiation is highly damaging for PSII, whose electron transport is affected both at the water oxidizing complex, and the binding site of the Q(B) quinone electron acceptor in a similar way to that caused by UV-B radiation. PMID- 11045724 TI - Photodetection with 5-Aminolevulinic acid-induced protoporphyrin IX in the rat abdominal cavity: drug-dose-dependent fluorescence kinetics. AB - In 75% of cases, ovarian carcinoma has already metastasized in the abdominal cavity at the time of diagnosis. For determination of the necessity for a supplementary therapy, in addition to surgical resection, it is important to localize and stage microscopical intraperitoneal metastases of the tumor. Intraperitoneal photodetection of tumor metastases is based on preferential tumor distribution of a fluorescent tumor marker. The time-dependent differences in drug concentration between tumor and normal (T/N) tissues can be used to visualize small tumors. We performed fluorescence measurements on abdominal organs and tumor in the peritoneal cavity of rats. 5-Aminolevulinic acid (ALA) induced protoporphyrin IX (PpIX) was used as the fluorescent marker. Three different drug doses (100, 25 and 5 mg/kg) were used and PpIX fluorescence profiles were followed up to 24 h after intravenous administration. Maximum T/N ratios were found 2-3 h after administration of ALA with all drug doses. A significant T/N tissue contrast was obtained for all abdominal organs tested after administration of 5 mg/kg. PMID- 11045725 TI - Increased eumelanin expression and tanning is induced by a superpotent melanotropin [Nle4-D-Phe7]-alpha-MSH in humans. AB - Seven normal volunteers (six males and one female) with tanning skin types III or IV (Fitzpatrick scale) were given 10 daily subcutaneous injections of a superpotent synthetic analog of alpha-melanocyte stimulating hormone (alpha-MSH) over two weeks. This agent, [Nle4-D-Phe7]alpha-MSH, also called Melanotan-I (MT I), was administered at a dose of 0.16 mg/kg/day (Monday-Friday), over a two week period. Tanning was measured serially using computerized light reflectance. This regimen induced tanning at 3 of 8 anatomic sites including the face, neck and forearm by comparison of baseline to (1) the end of the daily dosing period, (day 14), and (2) one week later, (day 21). Shave biopsies of the forearm taken at baseline and day 21 were analyzed by high performance liquid chromatography for eumelanin content which was measured as the permanganate oxidation product, pyrrole-2,3,5-tricarboxylic acid or PTCA. Pheomelanin content was measured as the hydroiodic acid digestion product, aminohydroxyphenylalanine (AHP). Eumelanin was also measured in the forehead skin samples of three subjects. The HPLC results show that mean (+/- SD) baseline eumelanin (PTCA) levels in forehead skin (n = 3) averaged 1.38 (+/- 0.87) ng/mg of wet skin tissue weight. Higher mean baseline levels of PTCA were detected in forearm skin (2.06 +/- 0.28 ng/mg wet weight, n = 7). One week after MT-I treatments ended, there was a mean (SD) 49% (+/- 17.6%) increase in forehead skin PTCA levels compared to baseline (P = 0.019, n = 3, by paired sample T-test). The mean (SD) increase in forearm skin PTCA levels was 98% (+/- 25.4%) over the same period (P = 0.003). In contrast, forearm pheomelanin expression following MT-I treatment did not significantly change from baseline. Overall, the MT-I regimen increased the eumelanin: pheomelanin ratio in forearm skin from 51:1 at baseline to 86:1 following MT-I (P = 0.054 by paired sample T test). These results show that the tanning induced by MT-I in the face and forearm is associated with a significant increase in the eumelanin content of the human skin. PMID- 11045726 TI - Scavenger-receptor targeted photodynamic therapy. AB - Covalent conjugation of a photosensitizer to a ligand that specifically recognized and internalized by a cell-surface receptor may be a way of improving the selectivity of photodynamic therapy (PDT). The class A Type-I scavenger receptor of macrophages, which among other ligands recognizes maleylated serum albumin and has a high capacity is a good candidate for testing this approach. Chlorin(e6) was covalently attached to bovine serum albumin to give conjugates with molar substitution ratios of 1:1 and 3:1 (dye to protein), and these conjugates could then be further modified by maleylation. A novel way of purifying the conjugates by acetone precipitation was developed in order to remove traces of unbound dye that could not be accomplished by size-exclusion chromatography. Conjugates were characterized by polyacrylamide gel electrophoresis and thin-layer chromatography. Photosensitizer uptake was measured by target J774 murine macrophage-like cells and nontarget OVCAR-5 human ovarian cancer cells, and phototoxicity was examined after illumination by a 660 nm diode laser by a tetrazolium assay. All of the purified conjugates were taken up by and after illumination killed J774 cells while there was only small uptake and no phototoxicity toward OVCAR-5 cells. The higher dye:protein ratio and maleylation of the conjugates both produced higher uptakes and lower survival ratios in J774 cells. The uptake and phototoxicity by J774 cells were decreased after incubation at 4 degrees C demonstrating internalization, and confocal microscopy with organelle-specific green fluorescent probes showed largely lysosomal localization. Uptake and phototoxicity by J774 cells could both be competed by addition of the scavenger receptor ligand maleylated albumin. These data show that scavenger receptor-targeted PDT gives a high degree of specificity toward macrophages and may have applications in the treatment of tumors and atherosclerosis. PMID- 11045727 TI - The binding characteristics and intracellular localization of temoporfin (mTHPC) in myeloid leukemia cells: phototoxicity and mitochondrial damage . AB - The state of aggregation of the photosensitizer meso-tetrahydroxyphenylchlorin (mTHPC) in both cell free and intracellular environment was elucidated by comparing its absorption and excitation spectra. In methanol, mTHPC existed as monomers and strongly fluoresced. In aqueous solutions such as phosphate-buffered saline (PBS), mTHPC formed nonfluorescent aggregates. Some portion of mTHPC monomerized in the presence of 10% fetal calf serum PBS. In murine myeloid leukemia M1 and WEHI-3B (JCS) cells, cytoplasmic mTHPC were monomeric. By using organelle-specific fluorescent probes, it was found that mTHPC localized preferentially at the mitochondria and the perinuclear region. Photodynamic treatment of mTHPC-sensitized leukemia cells caused rapid appearance of the apoptogenic protein cytochrome c in the cytosol. Results from flow cytometric analysis showed that the release of cytochrome c was especially pronounced in JCS cells, and well correlated with the extent of apoptotic cell death as reported earlier. Electron microscopy revealed the loss of integrity of the mitochondrial membrane and the appearance of chromatin condensation as early as 1 h after light irradiation. We conclude that rapid release of cytochrome c from photodamaged mitochondria is responsible for the mTHPC-induced apoptosis in the myeloid leukemia JCS and M1 cells. PMID- 11045728 TI - Singlet oxygen, but not oxidizing radicals, induces apoptosis in HL-60 cells. AB - Oxidizing species (OS), produced by photosensitization or derived from cytotoxic agents, activate apoptotic pathways. We investigated whether two different OS, formed at the same subcellular sites, have equivalent ability to initiate apoptosis in HL-60 cells. Our previous work showed that absorption of visible light by rose bengal (RB) produces singlet oxygen exclusively, whereas absorption of ultraviolet A produces RB-derived radicals in addition to singlet oxygen. Singlet oxygen, but not the RB-derived radicals, induced nuclear condensation and DNA fragmentation into nucleosome-size fragments in a dose dependent manner. In contrast, the RB-derived radicals caused greater lipid oxidation than singlet oxygen. These results indicate that different OS, produced at the same subcellular sites, do not have the same ability to induce apoptosis and that the ability of an OS to initiate lipid oxidation does not necessarily correlate with its ability to induce apoptosis. PMID- 11045729 TI - UV-enhanced expression of a reporter gene is induced at lower UV fluences in transcription-coupled repair deficient compared to normal human fibroblasts, and is absent in SV40-transformed counterparts. AB - UV irradiation enhances transcription of a number of cellular and viral genes. We have compared dose responses for alterations in expression from reporter constructs driven by the human and murine cytomegalovirus (CMV) immediate early (IE) promoters in cells from patients with deficiencies in nucleotide excision repair (complementation groups of xeroderma pigmentosum and Cockayne syndrome) following UV exposure, or infection with UV-damaged recombinant vectors. Results suggest that unrepaired damage in active genes triggers increased reporter activity from constructs driven by the CMV promoters in human fibroblasts. Similar to human fibroblasts, HeLa cells and cells from Li-Fraumeni syndrome patients (characterized by an inherited mutation in the p53 gene) also displayed an increase in reporter activity following UV exposure; however, this response was absent in all simian virus 40 (SV40)-transformed cell lines examined. This suggests that a pathway affected by SV40-transformation (other than p53) plays an essential role in UV-enhanced expression from the CMV IE promoter. PMID- 11045730 TI - Lack of correlation between DNA strand breakage and p53 protein levels in human fibroblast strains exposed to ultraviolet lights. AB - The contribution of DNA strand breaks accumulating in the course of nucleotide excision repair to upregulation of the p53 tumor suppressor protein was investigated in human dermal fibroblast strains after treatment with 254 nm ultraviolet (UV) light. For this purpose, fibroblast cultures were exposed to UV and incubated for 3 h in the presence or absence of l-beta-D arabinofuranosylcytosine (araC) and/or hydroxyurea (HU), and then assayed for DNA strand breakage and p53 protein levels. As expected from previous studies, incubation of normal and ataxia telangiectasia (AT) fibroblasts with araC and HU after UV irradiation resulted in an accumulation of DNA strand breaks. Such araC/HU-accumulated strand breaks (reflecting nonligated repair-incision events) following UV irradiation were not detected in xeroderma pigmentosum (XP) fibroblast strains belonging to complementation groups A and G. Western blot analysis revealed that normal fibroblasts exhibited little upregulation of p53 (approximately 1.2-fold) when incubated without araC after 5 J/m2 irradiation, but showed significant (three-fold) upregulation of p53 when incubated with araC after irradiation. AraC is known to inhibit nucleotide excision repair at both the damage removal and repair resynthesis steps. Therefore, the potentiation of UV-induced upregulation of p53 evoked by araC in normal cells may be a consequence of either persistent bulky DNA lesions or persistent incision associated DNA strand breaks. To distinguish between these two possibilities, we determined p53 induction in AT fibroblasts (which do not upregulate p53 in response to DNA strand breakage) and in XP fibroblasts (which do not exhibit incision-associated breaks after UV irradiation). The p53 response after treatment with 5 J/m2 UV and incubation with araC was similar in AT, XPA, XPG and normal fibroblasts. In addition, exposure of XPA and XPG fibroblasts to UV (5, 10 or 20 J/m2) followed by incubation without araC resulted in a strong upregulation of p53. We further demonstrated that HU, an inhibitor of replicative DNA synthesis (but not of nucleotide excision repair), had no significant impact on p53 protein levels in UV irradiated and unirradiated human fibroblasts. We conclude that upregulation of p53 at early times after exposure of diploid human fibroblasts to UV light is triggered by persistent bulky DNA lesions, and that incision-associated DNA strand breaks accumulating in the course of nucleotide excision repair and breaks arising as a result of inhibition of DNA replication contribute little (if anything) to upregulation of p53. PMID- 11045731 TI - Comparison of the pharmacokinetics and phototoxicity of protoporphyrin IX metabolized from 5-aminolevulinic acid and two derivatives in human skin in vivo. AB - Our novel approach was to compare the pharmacokinetics of 5-aminolevulinic acid (ALA), ALA-n-butyl and ALA-n-hexylester induced protoporphyrin IX (PpIX), together with the phototoxicity after photodynamic therapy (PDT) in human skin in vivo, using iontophoresis as a dose-control system. A series of four increasing doses of each compound was iontophoresed into healthy skin of 10 volunteers. The kinetics of PpIX metabolism (n = 4) and the response to PDT (n = 6) performed 5 h after iontophoresis, were assessed by surface PpIX fluorescence and post irradiation erythema. Whilst ALA-induced PpIX peaked at 7.5 h, highest PpIX fluorescence induced by ALA-n-hexylester was observed at 3-6 h and no clear peak was seen with ALA-n-butylester. With ALA-n-hexylester, more PpIX was formed after 3 (P < 0.05) and 4.5 h, than with ALA or ALA-n-butylester. All compounds showed a linear correlation between logarithm of dose and PpIX fluorescence/phototoxicity at 5 h, with R-values ranging from 0.87 to 1. In addition, the ALA-n-hexylester showed the tendency to cause greater erythema than ALA and ALA-n-butylester. Fluorescence microscopy (n = 2) showed similar PpIX distributions and penetration depths for the three drugs, although both ALA esters led to a more homogeneous PpIX localization. Hence, ALA-n-hexylester appears to have slightly more favorable characteristics for PDT than ALA or ALA-n-butylester. PMID- 11045732 TI - Induction of photolyase activity in wood frog (Rana sylvatica) embryos. AB - Rising ultraviolet-B (UVB, 280-320 nm) radiation has been proposed as a factor which may explain nonnormal amphibian population declines. Accordingly research has been directed toward estimating the photolyase activity of several amphibian species in order to predict a species' resilience to UV damage. Unfortunately, in spite of published research which demonstrated that the activity of one of the principal photorepair enzymes, photolyase, can be induced, these estimates did not address the potential for in vivo induction by environmental factors present in situ. We show here that wood frog (Rana sylvatica) embryos exposed to periods of ambient solar radiation (1) displayed significantly different photolyase activities from embryos exposed to equivalent periods of dark; and (2) were positively correlated with the UVB fluence received in vivo. Such results suggest that previous conclusions regarding the relationship between photorepair and population decline must be reevaluated. Estimating amphibian photorepair is a complicated process, and caution must be exercised when interpreting such data. PMID- 11045733 TI - Photoactivation of vascular iNOS and elevation of cGMP in vivo: possible mechanism for photovasorelaxation and inhibition of restenosis in an atherosclerotic rabbit models. AB - Recently, intravascular low-power red laser light (LPRLL) therapy has been proposed for the prevention of postangioplasty restenosis due to the observed inhibition of experimental neointimal formation. The objective of this study was to determine the impact of endoluminal LPRLL on vascular levels of inducible nitric oxide synthase (iNOS) and cyclic guanosine monophosphate (cGMP) to help define the mechanism of this effect. Eight atherosclerotic male adult New Zealand White rabbits weighing 4-6 kg were used in these studies. The iliac arteries were treated in separate zones with: (1) balloon inflation only; (2) laser illumination only; and (3) balloon inflation + laser illumination. An uninjured zone of the iliac artery served as a control. Laser irradiation (630 nm) was delivered to the vessel wall via a Cold laser Illuminator (Cook, Inc., Bloomington, IN), with a 3 mm-diameter balloon. Experiments demonstrated that vascular cGMP levels obtained immediately following treatment in the balloon only group was the lowest (0.29 +/- 0.05 pmol/mg protein) and significantly lower compared with the uninjured controls (1.01 +/- 0.07 pmol/ mg protein) (P < 0.001). In the laser only treated group cGMP levels were significantly increased (2.87 +/- 0.12 pmol/mg protein) compared with the uninjured control (P < 0.001) and the balloon only group (P < 0.001). Vascular cGMP levels in the balloon + laser group (2.09 +/0.07 pmol/mg protein) was also increased compared to the balloon only (P < 0.001) and control (P < 0.001) groups. Qualitative analysis of Western blot demonstrated that laser illumination induces iNOS. In contrast balloon dilatation did not induce iNOS. Balloon + laser treatment, however, tended to restore the expression of iNOS. Our study demonstrated that intravascular low dose laser irradiation induces iNOS and elevates vascular cGMP in an in vivo atherosclerotic rabbit model. PMID- 11045734 TI - The effects of approach and avoidance motor actions on the elements of creative insight. AB - The authors propose that the nonaffective bodily feedback produced by arm flexion and extension informs individuals about the processing requirements of the situation, leading to the adoption of differential processing styles and thereby influencing creativity. Specifically, the authors predicted that arm flexion would elicit a heuristic processing strategy and bolster insight processes, whereas arm extension would elicit a systematic processing strategy and impair insight processes. To test these predictions, the authors assessed the effects of these motor actions on 3 central elements of creative insight: contextual set breaking, restructuring, and mental search. As predicted, in 6 experiments, arm flexion, relative to arm extension, facilitated insight-related processes. In a 7th experiment, arm extension, relative to arm flexion, facilitated analytical reasoning, supporting a cognitive tuning interpretation of the findings. PMID- 11045735 TI - Counteractive self-control in overcoming temptation. AB - How do anticipated short-term costs affect the likelihood of engaging in an activity that has long-term benefits. Five studies investigated the factors that determine (a) how anticipated short-term costs elicit self-control efforts and (b) how self-control efforts eventually diminish the influence of short-term costs on behavior. The studies manipulated short-term costs (e.g., painful medical procedures) and assessed a variety of self-control strategies (e.g., self imposed penalties for failure to undergo a test). The results show that short term costs elicit self-control strategies for self rather than others, before rather than after behavior. when long-term benefits are important rather than unimportant and when the costs are moderate rather than extremely small or large. The results also show that the self-control efforts help people act according to their long-term interests. PMID- 11045736 TI - The sex-->aggression link: a perception-behavior dissociation. AB - Four studies suggest that priming may yield directionally different effects on social perception and behavior if perceptual and behavioral experiences with the stimulus diverge. This seems true for sex and aggression: Men are more likely to behave aggressively than women, whereas women are more likely to perceive aggressive behavior than men. Using a sequential priming paradigm, Study 1 demonstrates that a basic semantic link between sex and aggression exists for both genders. This link, however, has opposing behavioral and perceptual consequences for men and women. Studies 2 and 3 demonstrate that sex priming facilitates aggressive behavior only for men. Study 4 shows that only women perceive the ambiguously aggressive behavior of a male target person as more aggressive after sex priming. Thus, the perceptual and behavioral responses to sex priming are consistent with the experiences men and women typically have with sex and aggression. PMID- 11045737 TI - Perceived superiority in close relationships: why it exists and persists. AB - Two studies used a thought-listing technique to examine perceived superiority, or the inclination to regard one's own relationship as better than (and not as bad as) others' relationships. Consistent with the claim that this is a motivated phenomenon-and motivated in part by strong commitment-Study 1 revealed that (a) tendencies toward perceived superiority and (b) the commitment-superiority link are both strongest given psychologically threatening instructions and weakest given accuracy instructions (control instructions are intermediate). Consistent with the claim that this phenomenon serves a functional purpose, Study 2 revealed that earlier perceived superiority predicts later relationship status (persisted vs. ended) and increases over time in dyadic adjustment. Also, commitment accounts for unique variance in perceived superiority beyond self-esteem. PMID- 11045738 TI - The impact of past relationships on interpersonal behavior: behavioral confirmation in the social-cognitive process of transference. AB - This study extended research on transference in social perception (e.g., S. M. Andersen, I. Reznik, & L. M. Manzella, 1996) into the realm of social behavior by examining behavioral confirmation (e.g., M. Snyder, 1992) in transference. Each perceiver participated in a brief conversation with a naive target participant, who either did or did not appear to resemble the perceiver's own positively or negatively regarded significant other. Trained judges rated positive affect expressed in targets' behavior. As predicted, targets expressed more positive affect in their behavior when they allegedly resembled the perceiver's own positively versus negatively toned significant other, an effect not found in the control condition. This evidence demonstrates behavioral confirmation in transference, suggesting a means by which present relationships may resemble past ones. PMID- 11045739 TI - Downward comparison in everyday life: reconciling self-enhancement models with the mood-cognition priming model. AB - Two models concerning downward comparison are motivational; they predict that when people are unhappy, they make downward comparisons to self-enhance (e.g., Wills, 1981). In contrast, the affect-cognition priming model (Wheeler & Miyake, 1992) predicts that unhappy people make upward comparisons because negative affect makes mood-congruent comparisons more accessible. The authors propose that both motivational and accessibility factors influence social comparisons. A study of undergraduates' self-recorded everyday comparisons supported this view. In addition, results (a) pointed to motivational influences other than self enhancement and an accessibility influence other than mood-congruent priming, (b) suggested that motivated and unintended comparisons may differ somewhat in their susceptibility to motivational and accessibility influences, and (c) identified challenges to both self-enhancement and priming models. PMID- 11045740 TI - Motivation gains in performance groups: paradigmatic and theoretical developments on the Kohler effect. AB - In contrast to many demonstrations of social loafing, relatively few studies have documented group motivation gains. One such exception was O. Kohler's (1926, 1927) finding that team members working together did better at a taxing persistence task than would be expected from their individual performances, particularly when there was a moderate discrepancy in coworkers' capabilities. In Experiment 1, we developed a paradigm within which Kohler's overall motivation gain effect could be replicated, although the discrepancy in coworkers' capabilities did not moderate these motivation gains (after statistical artifacts were taken into account). Experiment 2 indicated that this motivation gain occurred under conjunctive but not under additive task demands, suggesting that the instrumentality of one's contribution to valued outcomes is a more likely explanation of the Kohler effect than social comparison processes. PMID- 11045741 TI - Intergroup emotions: explaining offensive action tendencies in an intergroup context. AB - Three studies tested the idea that when social identity is salient, group-based appraisals elicit specific emotions and action tendencies toward out-groups. Participants' group memberships were made salient and the collective support apparently enjoyed by the in-group was measured or manipulated. The authors then measured anger and fear (Studies 1 and 2) and anger and contempt (Study 3), as well as the desire to move against or away from the out-group. Intergroup anger was distinct from intergroup fear, and the inclination to act against the out group was distinct from the tendency to move away from it. Participants who perceived the in-group as strong were more likely to experience anger toward the out-group and to desire to take action against it. The effects of perceived in group strength on offensive action tendencies were mediated by anger. PMID- 11045742 TI - The psychological trade-offs of goal investment. AB - The primary goal of this research was to investigate the possibility that being very invested in goals has psychological trade-offs. Self-report methods were used in a concurrent study with college students (Study 1) and a longitudinal study with elementary school children (Study 2). The results of both studies provided support for the hypothesis that high goal investment has psychological trade-offs. Such investment was associated with positive emotions as well as with worrying, both concurrently and longitudinally. In addition, evidence for mediational mechanisms was provided: Perceptions of accomplishment accounted for the relation between goal investment and positive emotions; the link between goal investment and worrying was mediated by predictions that failure would be upsetting. The implications of these findings for distinguishing between depressive and anxiety symptoms are discussed. PMID- 11045743 TI - Stalking the perfect measure of implicit self-esteem: the blind men and the elephant revisited? AB - Recent interest in the implicit self-esteem construct has led to the creation and use of several new assessment tools whose psychometric properties have not been fully explored. In this article, the authors investigated the reliability and validity of seven implicit self-esteem measures. The different implicit measures did not correlate with each other, and they correlated only weakly with measures of explicit self-esteem. Only some of the implicit measures demonstrated good test-retest reliabilities, and overall, the implicit measures were limited in their ability to predict our criterion variables. Finally, there was some evidence that implicit self-esteem measures are sensitive to context. The implications of these findings for the future of implicit self-esteem research are discussed. PMID- 11045744 TI - Emotional experience in everyday life across the adult life span. AB - Age differences in emotional experience over the adult life span were explored, focusing on the frequency, intensity, complexity, and consistency of emotional experience in everyday life. One hundred eighty-four people, age 18 to 94 years, participated in an experience-sampling procedure in which emotions were recorded across a 1-week period. Age was unrelated to frequency of positive emotional experience. A curvilinear relationship best characterized negative emotional experience. Negative emotions declined in frequency until approximately age 60, at which point the decline ceased. Individual factor analyses computed for each participant revealed that age was associated with more differentiated emotional experience. In addition, periods of highly positive emotional experience were more likely to endure among older people and periods of highly negative emotional experience were less stable. Findings are interpreted within the theoretical framework of socioemotional selectivity theory. PMID- 11045745 TI - Personality, emotional experience, and efforts to control emotions. AB - Three converging, multimethod studies examined personality and emotional processes. Study 1 (N = 321) examined links among sex, personality, and expectations for emotional events. In Study 2, participants (N = 468) described contents of emotionally evocative slides to a partner (either a friend or a stranger). Participants reported their emotional experience, efforts to control emotion, and the anticipated reactions of their partners. Structural modeling of self-report data and analyses of observational data indicated that Agreeableness and sex were significant predictors of emotional experience and of efforts to control emotion. Study 3 (N = 68) replicated and extended the two previous studies using psychophysiological methods to examine responses to positively and negatively charged emotional materials. Outcomes are discussed in terms of processes underlying the five-factor structural dimension of Agreeableness and links to emotional self-regulation. PMID- 11045746 TI - Scandinavia's lessons to the world of public health. PMID- 11045747 TI - Neck or shoulder pain and low back pain in Finnish adolescents. AB - The aim of this study was to investigate the prevalence and determinants of self reported neck or shoulder pain (NSP) and low back pain (LBP) among 12-18-year olds. A questionnaire was mailed to a nationally representative sample of 11,276 12-, 14-, 16- and 18-year-olds in 1991. The response rate was 77%. NSP was perceived at least once a week by 15% of 12-18-year-olds and LBP by 8%. Both symptoms were more prevalent among girls than among boys, and the prevalence increased with age. Among the determinants investigated, the number of perceived psychosomatic symptoms had the strongest association with NSP and LBP. Our study confirmed the co-morbidity of NSP and LBP, and indicated that NSP is more frequent than believed among 16-18-year-old girls. The strong association of psychosomatic symptoms with NSP and LBP suggests that the latter two pain states could be more psychosomatic than nociceptive in character. PMID- 11045748 TI - Visiting the cinema, concerts, museums or art exhibitions as determinant of survival: a Swedish fourteen-year cohort follow-up. AB - The aim of this study was to ascertain the possible influence of attending various kinds of cultural events or visiting cultural institutions as a determinant of survival. A cohort of individuals aged 25-74 years from a random sample were interviewed by trained non-medical interviewers in 1982 and 1983. The interviews covered standard-of-living variables. Our independent variables covered visiting cultural institutions and attendance at cultural events, reading books or periodicals, and music making. The non-response rate was about 25%. The cohort was followed with respect to survival for 14 years up to 31st December 1996. The background covariates that were used for control purposes were age, sex, cash buffer, educational standard, long-term disease, smoking, and physical exercise. Our setting was the Swedish survey of living conditions among the adult Swedish population aged 25-74 years. About 10,609 individuals were interviewed in 1982 and 1983. The outcome measure was survival until 31st December 1996. In all, 916 men and 600 women died during this period. We found a higher mortality risk for those people who rarely visited the cinema, concerts, museums, or art exhibitions compared with those visiting them most often. The significant relative risks ranging between RR 1.14 (95% CI. 1.01-1.31) of attending art exhibitions, and RR 1.42 (CI. 1.25-1.60) of attending museums, when adjusting for the nine other variables. Visits to the cinema and concerts gave significant RR in between. We could not discern any beneficial effect of attending the theatre, church service or sports event as a spectator or any effect of reading or music making. Our conclusion is that attendance at certain kinds of cultural events may have a beneficial effect on longevity. PMID- 11045749 TI - Minority status and perceived health: a comparative study of Finnish- and Swedish speaking schoolchildren in Finland. AB - Results of earlier studies suggest that the health of the Swedish-speaking minority in Finland is better than that of the sociodemographically similar Finnish-speaking population. The causes of differences are unknown. The main aim of the study referred was to investigate whether differences in perceived health according to linguistic group were present in a nation-wide representative sample (n=5,230) of schoolchildren aged 11, 13 and 15 years. A further aim was to determine whether differences could be attributed to socioeconomic background, social relationships or health behaviour. The study is part of the international Health Behaviour in School-aged children (HBSC) survey. The perceived health of Swedish-speaking children (n=1,699) proved to be better than that of Finnish speaking children (n=3,531). In multivariate logistic regression models the differences could not be attributed to underlying associations with any variable studied. The health advantage of Swedish-speaking children essentially could not be related to known risk factors. PMID- 11045750 TI - Risk of childhood injury: predictors of mothers' perceptions. AB - BACKGROUND AND OBJECTIVES: Safety education often targets parental risk perception. Predictors of risk perception, however, are not well known, thus limiting the feasibility of effective safety education. Accordingly, in this study, a range of predictors of maternal risk perception were examined. METHODS: A random sample of 870 mothers in northern Sweden was included in the study. Three different questionnaires, with scenarios of a burn injury, a bicycle injury in the home environment, and a bicycle injury in traffic, were completed by the subjects. Multiple linear regression models tested the possible influence of causal attributions, normative beliefs, and sociodemographic and behaviour related variables on mothers' risk perception. RESULTS: Only 14-23% of the variance in mothers' risk perception could be explained by the multivariate models. Causal attribution to the child was found to be the most important predictor of maternal risk perception. CONCLUSION: Present theoretical models give few clues about how to design educational models that might influence risk perception. To make safety education more effective, other modifiable factors that influence parental safety behaviour, such as subjective norms and self efficacy, might be better targets. PMID- 11045751 TI - Socioeconomic differences in health expectancy in Denmark. AB - BACKGROUND: Social differences in mortality rates reported in Denmark gave rise to the present study of health expectancy in different socioeconomic groups. METHODS: Data on health status and occupation were derived from Health Interview Surveys. Information on occupation and deaths is register data. Health expectancy in each socioeconomic group was calculated using Sullivan's method. RESULTS: Among 30-year-old men, high-level salaried employees had the longest expected lifetime in perceived good health, 41 years, which amounts to 89% of life expectancy, compared to 34 years (73%) for farmers, 32 years (73%) for unskilled workers, and 19 years (56%) for economically inactive men. Expected lifetime in perceived good health for high-level salaried female employees from age 30 was 46 years (91% of life expectancy). The lowest was found for assisting spouses, 36 years (71%) and economically inactive women, 25 years (56%). Large differences were also found when data on long-standing illness were used. CONCLUSION: The differences between socioeconomic groups are greater for health expectancy than for life expectancy. PMID- 11045752 TI - Socioeconomic differences in smoking cessation: the role of social participation. AB - OBJECTIVES: The aim of this study was to investigate whether psychosocial resources explain socioeconomic differences in smoking cessation and its maintenance. METHODS: A subpopulation of 11,837 individuals from the Malmo Diet and Cancer Study interviewed in 1992-94, age range 45-64 years, was investigated in this cross-sectional study. A multivariate logistic regression model was used to assess relative risks of having stopped smoking, adjusting for age, country of origin, previous/current diseases, and marital status. RESULTS: An odds ratio of 1.9 (1.4-2.5; 95% CI) for men and 2.0 (1.4-2.7; 95% CI) for women of having stopped smoking was found for higher non-manual employees when compared with unskilled manual workers. A decrease in these odds ratios was found when social participation was introduced into the model. The other three social network and social support variables were non-significant. CONCLUSION: High social participation is a predictor of maintenance of smoking cessation. It seems possible to interpret parts of the socioeconomic differences in smoking cessation and its maintenance as a consequence of differing social network resources and social capital between socioeconomic groups. PMID- 11045753 TI - Violence in an urban community: a population-based interview study. AB - BACKGROUND: This study aimed to estimate the proportion of an unselected population in an urban community (Bergen, Norway) that was exposed to physical violence during the preceding year, and to determine the proportion seeking medical treatment and pressing legal charges. METHOD: A structured telephone interview was performed as part of monthly opinion polls. RESULTS: During a one year period in 1997/1998, 3,005 residents of Bergen were interviewed. Of these, 41 (1.4%) had been exposed to physical violence during the preceding year; 10 (24%) of them had sought medical treatment, and 16 (39%) had pressed legal charges. The majority of the victims had been treated at Bergen Accident and Emergency Department. The interviewees knew about a total of 347 other people who had been assaulted during the preceding year. Of these assault victims, 224 (65%) sought medical treatment and 181 (52%) pressed legal charges, according to the respondents. CONCLUSION: A high proportion of the assault victims did not seek medical treatment and did not press legal charges. PMID- 11045754 TI - Sex differences in mortality in Denmark during half a century, 1943-92. AB - OBJECTIVE: The emphasis of this study is on the relative mortality of 45-74-year old men and women in Denmark in 1943-92, following economic and political changes that have affected the social meaning of gender over the last 50 years, and which have diminished former sex differences in health behaviour. METHODS: Sex ratios of total mortality and mortality from major non-sex-specific causes of death were calculated on computerized mortality data from the Danish National Cause of Death Register that covers all deaths in Denmark since 1943. RESULTS: In the early 1940s the sex ratio of all-cause mortality was low, 1.0-1.1, it increased to a peak level in the late 1970s and early 1980s, but has since decreased due to an increase in female mortality and a more favourable trend in male mortality. CONCLUSION: Gender equality, employment, and economic autonomy may have beneficial health effects on both men and women, but the effects are inconsistent. The trend in smoking is the major explanatory factor for the more recent trends in gender differentials in mortality in Denmark. PMID- 11045755 TI - Barriers within the health care system to dealing with sexualized violence: a literature review. AB - The aim of this study was to review the literature about possible barriers to recognition and intervention regarding women exposed to sexualized violence, in their interactions with the health care system. The barriers, as reported by the health care staff, were: lack of education; the stereotype of a "typical battered woman"; too close identification with the victim/abuser; time constraints; fear of offending the victim/abuser; and feelings of hopelessness and non responsibility. The barriers, as reported by the victims, were: negative experiences of and structural limitations within the health care system; fear of retaliation from the abusive partner; and psychological effects of the normalization process. We conclude that the barriers within the health care sector have to be dealt with on three different levels: the structural level in order to diminish male power in society; the organizational level in order to initiate screening and to allow the staff time for dealing with the victims; and on the individual level, health care staff need to acquire the knowledge and skills to enable them to address sexualized violence. PMID- 11045756 TI - Is sitting-while-at-work associated with low back pain? A systematic, critical literature review. AB - OBJECTIVES: To present a critical review and evaluate recent reports investigating sitting-while-at-work as a risk factor for low back pain (LBP). METHODS: The Medline, Embase and OSH-ROM databases were searched for articles dealing with sitting at work in relation to low back pain for the years 1985-97. The studies were divided into those dealing with sitting-while-working and those dealing with sedentary occupations. Each article was systematically abstracted for core items. The quality of each article was determined based on the representativeness of the study sample, the definition of LBP, and the statistical analysis. RESULTS: Thirty-five reports were identified, 14 dealing with sitting-while-working and 21 with sedentary occupations. Eight studies were found to have a representative sample, a clear definition of LBP and a clear statistical analysis. Regardless of quality, all but one of the studies failed to find a positive association between sitting-while-working and LBP. High quality studies found a marginally negative association for sitting compared to diverse workplace exposures, e.g. standing, driving, lifting bending, and compared to diverse occupations. One low quality study associated sitting in a poor posture with LBP. CONCLUSIONS: The extensive recent epidemiological literature does not support the popular opinion that sitting-while-at-work is associated with LBP. PMID- 11045757 TI - Current trends and new approaches in the management of diabetes mellitus. AB - Current trends in the management of type 2 diabetes mellitus, based on the 20 year United Kingdom Prospective Diabetic Study, include intensive treatment to control the blood glucose level and blood pressure in order to prevent or delay microvascular and cardiovascular complications. In the new millennium, type 2 diabetes will become epidemic in developing countries. If diabetes were to develop in 10% of the 1.2 billion population of China, the expense of intensive treatment would be immense. Laboratory tests are useful for detecting risk factors before the onset of the disease and convincing the general public to take preventive measures. Glucose tolerance testing is one of these tests. When glucose tolerance is impaired, 25% of beta-cell function is lost. Determining the plasma proinsulin level is another useful evaluation; impaired glucose tolerance accompanied by increased plasma proinsulin level is indicative of an enhanced risk that type 2 diabetes will develop within 5 years. Educating the public about eating a healthy diet and exercising may prevent the development of diabetes and thereby reduce the global prevalence of type 2 diabetes. PMID- 11045758 TI - Effect of aerobic training on diabetic nephropathy in a rat model of type 2 diabetes mellitus. AB - The effect of aerobic exercise intervention on the renal functional and ultrastructural changes associated with diabetes mellitus were studied in the obese Zucker rat, a rat model of type 2 diabetes. The obese Zucker rats began training at 18 wk of age (n=8) and were compared to obese sedentary controls (n=12) and lean sedentary nondiseased littermates (n=10). Body weight, kidney weight, serum creatinine, urine creatinine, creatinine clearance, urine IgG, urine IgG/creatinine ratio, urine total protein, urine albumin, urine albumin/creatinine ratio, glycated hemoglobin, serum fructosamine, fasting serum glucose, serum insulin, serum total cholesterol, serum triglycerides, blood pressure, and morphometric analyses of cortical glomeruli by light microscopy and electron microscopy were performed to evaluate renal function, structure, and metabolic control. The exercise training consisted of treadmill running, 5 da/wk for 1 hr/da. Exercise intervention lowered the body weight (p <0.05), reduced the percentage of glycated hemoglobin (p <0.05), and diminished the urine albumin concentration (p <0.05), compared to the obese sedentary controls. Exercise intervention did not significantly affect morphometric indices of renal ultrastructure. This study shows that aerobic exercise intervention significantly improved metabolic control and reduced albuminuria in a rat model of type 2 diabetes. PMID- 11045759 TI - Effects of iron overload on the immune system. AB - Iron and its binding proteins have immunoregulatory properties, and shifting of immunoregulatory balances by iron excess or deficiency may produce severe, deleterious physiological effects. Effects of iron overload include decreased antibody-mediated and mitogen-stimulated phagocytosis by monocytes and macrophages, alterations in T-lymphocyte subsets, and modification of lymphocyte distribution in different compartments of the immune system. The importance of iron in regulating the expression of T-lymphocyte cell surface markers, influencing the expansion of different T-cell subsets, and affecting immune cell functions can be demonstrated in vitro and in vivo. The poor ability of lymphocytes to sequester excess iron in ferritin may help to explain the immune system abnormalities in iron-overloaded patients. Iron overload as seen in hereditary hemochromatosis patients enhances suppressor T-cell (CD8) numbers and activity, decreases the proliferative capacity, numbers, and activity of helper T cells (CD4) with increases in CD8/CD4 ratios, impairs the generation of cytotoxic T cells, and alters immunoglobulin secretion when compared to treated hereditary hemochromatosis patients or controls. A correlation has recently been found between low CD8+ lymphocyte numbers, liver damage associated with HCV positivity, and severity of iron overload in beta-thalassemia major patients. Iron overload, with its associated increases of serum iron levels and transferrin saturation, may cause a poor response to interferon therapy. Iron overload with hyperferremia is associated with suppressed functions of the complement system (classic or alternative types). High plasma ferritin content in patients with chronic, diffuse diseases of the liver (cirrhosis, chronic hepatitis), beta-thalassemia major, dyserythropoiesis, and hereditary hemochromatosis may induce the development of anti-ferritin antibodies with the production of circulating immune complexes. Increased body stores of iron in various clinical situations may tip the immunoregulatory balance unfavorably to allow increased growth rates of cancer cells and infectious organisms, and complicate the clinical management of preexisting acute and chronic diseases. PMID- 11045760 TI - Immunologic effects of gliotoxin in rats: mechanisms for prevention of autoimmune diabetes mellitus. AB - Various fungal products, such as gliotoxin (GT), have immunomodulating activity, a fact exploited previously by our group for prevention of autoimmune diabetes mellitus in BB/Wor rats. To understand better the immunologic effects in GT treated rats, splenocytes from 65-day-old prediabetic diabetes-prone rats were phenotypically characterized after chronic treatment with GT. A parallel study examined the direct effects of GT on splenocyte preparations incubated with the mycotoxin. In vitro treatment of splenocytes with GT revealed relative decreases in CD4+ and increases in CD8+ T-cell subsets, whereas in vivo treatment with GT did not result in detectable alterations in relative CD4+ and CD8+ cell subsets. We were unable to show significant effects on NK cells or MHC class II cells. However, in vitro and in vivo GT treatments significantly enhanced the detectable RT6 surface marker, a key regulatory element in autoimmune diabetes pathogenesis. This study showed that GT selectively affects certain lymphocyte subsets, possibly through the mechanism of apoptosis, which was increased in vivo as well as in vitro. PMID- 11045761 TI - Cell typing the sensitized transfusion-dependent patient. AB - Extended red cell typing is required for the management of transfusion-dependent patients to confirm the identity of suspected alloantibodies or determine the specificity of potential additional antibodies that may be formed in the future. Typing may be complicated by the presence of circulating allogeneic cells or a positive direct antiglobulin test. Phenotyping such individuals by hemagglutination is dependent on the separation of a reticulocyte-enriched fraction by differential centrifugation. Flow cytometric typing of reticulocytes is also possible. The effectiveness of these techniques is limited in those who are heavily transfused or have low reticulocyte counts. Heavily transfused patients with sickle cell anemia may be typed, however, following hypotonic lysis of allogeneic cells. In patients with a positive direct antiglobulin test, sensitized cells are usually typed with either direct agglutinating antisera and/or IgG antisera following elution of the autoantibody. Inactivation of some antigens during the elution process or the lack of some antisera specificities limit such typing. By designing appropriate oligonucleotide primers, polymerase chain reaction (PCR) amplification of target gene sequences for most blood group systems and the identification of a large number of their allelic specificities is now possible. Peripheral blood leukocytes can be used as the DNA source. Restriction fragment length polymorphism determination is widely adopted for the identification of allelic specificity of the amplified target sequence. Alternate strategies, including allele-specific PCR, are often employed if the genetic basis of the polymorphism is more complex than a single nucleotide substitution, or if it does not create or ablate a restriction endonuclease cleavage site. These techniques may permit genotyping of sensitized transfusion-dependent patients, and can improve transfusion safety and efficacy. PMID- 11045762 TI - Novel mutations, including a novel G659A missense mutation, of the FUT1 gene are responsible for the para-Bombay phenotype. AB - Para-Bombay phenotype, with an estimated incidence of 1 in 8000 in Taiwanese residents based on serological analysis, is caused by aberrant alpha(1,2) fucosyltransferase function and hence diminished H-antigen synthesis. In an individual with para-Bombay phenotype, DNA sequencing revealed two missense mutations previously reported C658T mutation and a novel G659A mutation. Haplotype analysis with restriction enzyme digestion showed that the two mutations are located on opposing alleles of the H (FUT1) gene and lead to compound heterozygosity. Since no other known genetic changes were evident, it appears that the new missense mutation, G659A, is deleterious to the alpha(1,2) fucosyltransferase function encoded by the H (FUT1) gene. PMID- 11045763 TI - The proportion of hybrid heterodimers in homozygous or doubly heterozygous beta chain variant hemoglobinopathies associated with alpha chain hemoglobin variants. AB - Four alpha genes exist on chromosome 16, but one or more of these genes can be deleted in association with Hemoglobin (Hb)G-Philadelphia in cis to alpha thalassemia-2 in African-Americans. Therefore, the proportion of HbG-Philadelphia in HbG heterozygotes is trimodal at about 25% for alphaGalpha/alpha alpha, 33% for alphaG-/alpha alpha, and 50% for alphaG-/alpha alpha in patients with HbA. Those who are homozygous or doubly heterozygous for beta chain variants (betaX2 or betaXbetaY) have neither HbA nor the alpha chain variant (alphaX2 betaA2), but have hybrid heterodimers (alphaX2 betaX2). The proportion of hybrid heterodimers here should also be trimodal mirroring alpha gene status. Eleven patients were identified: 4 with Hb SSG, 3 with Hb SCG, and 1 each with Hb OCG, HbSSMontgomery, HbSSChicago, and HbSSBourmedes. Heterodimer proportions were: 43.3 +/- 1.5, 33.5 +/- 2.3, and 15.8 +/- 1.1% for 2, 3, and 4 respective alpha genes which had been studied in 8/11 of the patients (r = 0.98), implying that the prime determinant of the proportion of hybrid heterodimers in this patient group is the number of functional alpha genes. PMID- 11045764 TI - Adenoviral-mediated gene therapy with Ad5CMVp53 and Ad5CMVp21 in combination with standard therapies in human breast cancer cell lines. AB - Our objective was to determine the efficacy of adenoviral-mediated gene therapy with wild-type p53 or p21 in human breast cancer cells and investigate interactions with radiation and chemotherapy. Two human breast cancer cell lines, MDA-MB-231 and MDA-MB-435, both with p53 mutations, were transduced with adenoviral vectors containing wild-type p53 (Ad5CMV-p53) or p21/WAF1/Cip1 (Ad5CMV p21), and the effects on growth were determined. Infection was combined with low dose (1.4 - 3.7 Gy) irradiation to see if this would improve transduction efficiency and enhance numbers of cells killed. Transduction with either vector resulted in expression of p21WAF1/cip1 and growth inhibition, although Ad5CMV-p53 transduction produced greater growth inhibition than did Ad5CMV-p21. The cell lines differed in sensitivity to the vectors. The Ad5CMV-p53 vector in a multiplicity of infection (MOI) of 125 resulted in 50% to 80% inhibition of MDA MB-231, while MOI 250 of the same vector resulted in 27% inhibition of MDA-MB 435. Infection with Ad5CMV-p21 produced modest growth inhibition in both cell lines (< or = 40% at MOI 200), although protein expression was detected at lower viral doses. Low dose gamma-irradiation (1.4 to 3.7 Gy) was used to try and improve the rate of gene transfer. Modest increases in transduction efficiency and duration of expression of a vector containing beta-galactosidase occurred in irradiated breast cancer cells. Radiation 24 hr before transduction with Ad5CMV p53 increased the proportions of apoptotic MDA-MB-231 cells. The cells transduced with Ad5CMV-p21 were arrested in G1, yet when they were irradiated before adenoviral transduction, the overexpression of p21 protected the cells from the cytotoxic effects of the radiation. Clonogenic assays showed that Ad5CMV-p21 reduced the sensitivity of MDA-MB-231 to VP-16 and paclitaxel. Combining these drugs with Ad5CMV-p53 did not consistently or significantly decrease clonogenic survival. PMID- 11045765 TI - How effective are screening tests for microalbuminuria in random urine specimens? AB - The effectiveness of four urine screening tests-microalbumin (MAlb), total protein (TProt), total protein/creatinine ratio (TProt/Cr R), and dipstick (DPalb) test for albumin-were evaluated for the detection of MAlb in random urine specimens. The following criteria were used to assess the effectiveness of each urine screening test: 100% specificity (no false positive results); cost effectiveness; rapidity and ease of performing the screening test; and increased laboratory efficiency. A "gold standard" for presence of MAlb in random urine samples was defined as a microalbumin/creatinine ratio (MAlb/Cr R) of > or = 30 mg/g. The least costly urine screening test was the DPalb, which, if assigned a value of 1.0, allowed a cost ranking order for the screening tests-DPalb (1.0) < urine TProt (1.03) < urine TProt/Cr R (2.1) < urine MAlb (7.0). Two hundred urine samples from diabetic inpatients and outpatients were tested. Only two screening tests--MAlb and DPalb--achieved 100% specificity without increasing laboratory costs (small net savings), whereas the other two screening tests--TProt and TProt/Cr R-only achieved 100% specificity with increased laboratory costs. Theoretical prevalence rate analysis showed that urine MAlb screening would be effective at all prevalence rates for overt nephropathy. TProt and DPalb urine screening testing would be most effective in populations with prevalence rates of > or = 15% for overt nephropathy. The TProt/Cr R ratio would only be effective in populations with prevalence rates of > or = 30%. Of the four urine screening tests, only DPalb would significantly streamline the process of measuring urine MAlb. The dipstick test is inexpensive, easy and rapid to perform, does not delay measuring the ratio, since there is no wait for the screening test result, and can be used by referring laboratories to screen urine specimens before they are submitted to a central laboratory, thereby reducing laboratory workload. PMID- 11045766 TI - Troponin I: an update on clinical utility and method standardization. AB - Cardiac troponin I (cTnI) is now widely recognized as one of the preeminent biochemical markers for the diagnosis of myocardial injury. The biochemical specificity of this biomolecule for cardiac tissue has forced a reevaluation of the diagnostic criteria for non-Q-wave acute myocardial infarction, unstable angina, acute coronary artery disease, and minor myocardial injury. Further, its use by clinicians has revolutionized the way that chronic and acute heart diseases are both diagnosed and managed. Unfortunately, the standardization of cardiac troponin I assays is problematic. Up to 20-fold variation of serum cTnI mass determinations may be observed for a given patient sample when measured by different assay systems. As a result, significant ambiguity often exists in the clinical interpretation of serum cTnI concentrations. Recent efforts have been directed toward the biochemical standardization of cTnI assays. However, the heterogeneous nature and biochemical complexity of the serum forms of cTnI and differences of the epitope recognition by the various methods have hindered the harmonization of serum cTnI assays. PMID- 11045767 TI - Serum lipid concentrations change with serum alkaline phosphatase activity during pregnancy. AB - To investigate the relationship between serum lipids and alkaline phosphatase during normal pregnancy, we measured triglyceride, total cholesterol, HDL cholesterol, and LDL-cholesterol concentrations and alkaline phosphatase activity in serum samples from 546 apparently healthy pregnant, postpartum, and nonpregnant women. Serum HDL-cholesterol levels did not change significantly during pregnancy, but serum triglyceride, total cholesterol, LDL-cholesterol, and alkaline phosphatase levels increased gradually as pregnancy proceeded, reached maximum values in the third trimester, and returned to nonpregnant levels by 20 24 wk postpartum. The serum alkaline phosphatase activity averaged 2.1-fold higher in the late third trimester than in the first trimester; the serum triglyceride concentration averaged 2.3-fold higher in the late third trimester than in the first trimester. Compared to the peak values during pregnancy, serum alkaline phosphatase activity averaged 45% lower and serum triglyceride level averaged 47% lower at 12-16 wk postpartum. The serum alkaline phosphatase activity was correlated with the serum concentrations of total cholesterol (r = 0.68, p < 0.01) and triglyceride (r = 0.71, p < 0.01). In short, this study shows that serum triglyceride and total cholesterol levels change in parallel with serum alkaline phosphatase activity during and after normal pregnancy. PMID- 11045768 TI - Chromatographic measurements of memoglobin A2 in blood samples containing sickle hemoglobin. PMID- 11045769 TI - Introduction: narrative in psychotherapy: the emerging metaphor. PMID- 11045770 TI - Cognitive narrative psychotherapy: research foundations. AB - There has been an increased interest in the study of language processes in psychotherapy. More recently, research and theoretical formulations of the therapeutic process suggested that we must move from the microscopic study of verbal modes to a macroscopic approach in which these modes are organized into narratives. Narratives are conceived, in this perspective, as the basic instruments for meaning making. In this article the research on narrative processes in psychotherapy is reviewed and discussed in terms of its implications for the theory and practice of cognitive narrative psychotherapy. Additionally some of the main data coming from research projects on cognitive narrative psychotherapy are presented. PMID- 11045771 TI - Self-narrative as meaning construction: the dynamics of self-investigation. AB - Starting from the metaphor of the person as a motivated storyteller, a theory of meaning construction and reconstruction is presented. Two motives are assumed to be particularly influential in the process of meaning construction: The striving for self-enhancement and the longing for contact and union with somebody or something else. A self-confrontation method is discussed and illustrated, enabling clients to perform, in close cooperation with the psychotherapist, a self-investigation on the content and organization of their personal meaning units. The method represents a gradual transition between assessment and change. Three functions of the method are discussed: assessment, process promotion, and evaluation. These functions are illustrated with a diversity of clinical phenomena: the finding of a central theme in the client's self-narrative, the experience of hopelessness and helplessness, the organized nature of depression, and the construction of a scenario for emerging from a depressive state. Finally, the multivoiced and dialogical nature of the self is illustrated by the dream of a murderer who was perceived by the client as both inside and outside the self. Special attention is given to the shifting boundaries between self and nonself. PMID- 11045772 TI - Some functions of narrative in the assimilation of problematic experiences. AB - We present three current, complementary formulations of the assimilation model: (a) the schema formulation, based on cognitive developmental concepts: (b) the voices formulation, in which assimilation is understood as the construction of a meaning bridge between active internal voices; and (c) the cognitive science formulation, which uses cognitive concepts of memory types to understand the failure of memory in cases of warded-off and avoided experiences. These views of assimilation are used to understand the varied functions that narratives (stories about real or imagined events outside of therapy) may play in psychotherapy, including narratives that avoid encounters with threatening material, narratives that approach such material indirectly or symbolically, narratives by which clients reexperience trauma, and narratives that help construct a mature understanding. PMID- 11045773 TI - Reliability and base rates of interpersonal themes in narratives from psychotherapy sessions. AB - We present an analysis of the reliability and base rates of the interpersonal contents of narratives told by patients in psychotherapy. Trained judges rated two samples, including 60 opiate-dependent patients in cognitive or psychodynamic therapy and 72 depressed patients in cognitive or interpersonal therapy. Using a comprehensive system based upon a circumplex model and involving 104 separate categories, we found that most categories of interpersonal behavior could be rated reliably. Potential problem categories were identified and strategies for increasing reliability are discussed. In particular, categories related to the concept of the introject (what the self does to the self) had low reliability. An analysis of the base rates of interpersonal themes revealed that issues related to autonomy/ assertion were most prevalent, although some differences between the two samples were evident. The implications of the results for research on narratives and models of psychotherapy are discussed. PMID- 11045774 TI - Forming a story: the health benefits of narrative. AB - Writing about important personal experiences in an emotional way for as little as 15 minutes over the course of three days brings about improvements in mental and physical health. This finding has been replicated across age, gender, culture, social class, and personality type. Using a text-analysis computer program, it was discovered that those who benefit maximally from writing tend to use a high number of positive-emotion words, a moderate amount of negative-emotion words, and increase their use of cognitive words over the days of writing. These findings suggest that the formation of a narrative is critical and is an indicator of good mental and physical health. Ongoing studies suggest that writing serves the function of organizing complex emotional experiences. Implications for these findings for psychotherapy are briefly discussed. PMID- 11045775 TI - The narrative processes coding system: research applications and implications for psychotherapy practice. AB - The Narrative Processes model is focused on the strategies and processes by which a client and therapist transform the events of everyday life into a meaningful story that both organizes and represents the client's sense of self and others in the world. Some investigators have elected to use clients' within session descriptions of relationship events or micronarratives as their unit of narrative analysis. In contrast, we are centrally interested in the development of the macronarrative framework in which the singular events described in a therapy relationship-micronarratives-come to be articulated, experienced, and linked together in such a way that the client's sense of his or her life story-in essence, the sense of self-may be transformed at the conclusion of the therapeutic relationship. The following paper details the Narrative Processes theory of therapy and the coding system that has been developed to identify and evaluate empirically key components of the model. Findings emerging from the analyses of successful psychotherapy dyads are described and the implications for future research and practice are discussed. PMID- 11045776 TI - Psychometric properties of the frost multidimensional perfectionism scale in a clinical anxiety disorders sample. AB - The purpose of this study was to examine the factor structure of the Frost Multidimensional Perfectionism Scale (MPS-F; Frost, Marten, Laharte, & Rosenblate, 1990). Although perfectionism is thought to contribute to the development of psychopathology and the MPS-F is gaining popularity for use in assessing perfectionism in clinical samples, to date the factor structure has not been examined in a clinical sample. Three hundred and twenty-two individuals diagnosed with an anxiety disorder using the SCID for DSM-IV and 49 nonclinical controls completed the MPS-F as well as a measure of perfectionism (MPS-H) developed by Hewitt and Flett ( 1991 ). Analyses suggested that the MPS-F has similar psychometric properties in clinical samples to those in nonclinical samples, and factors very similar to those observed by Frost et al. (1990) could be extracted. A 3-factor solution appeared more appropriate for statistical reasons, and the 3 scales based on these factors distinguished among diagnostic groups in a manner similar to scales based on the 6-factor solution in past research. Results were discussed in terms of the potential utility of a 3-factor solution and in terms of the general construct of perfectionism and the distinction between nonpathological high performance standards and neurotic perfectionism. PMID- 11045777 TI - Psychological characteristics of candidates for liver transplantation: differences according to history of substance abuse and UNOS listing. United Network for Organ Sharing. AB - Liver transplantation for patients with a history of substance abuse remains controversial. Resumption of heavy alcohol use postoperatively is a threat to long-term survival, but recidivism among transplanted alcoholics is reportedly low. An argument against psychological evaluation prior to transplantation revolves around the speculation that candidates will attempt to portray themselves as more desirable prior to listing with UNOS (United Network for Organ Sharing). This study measured psychological distress, coping styles, optimism, selected personality features, and perception of family environment among 73 U.S. military veterans who were candidates for liver transplantation. Candidates with positive histories of substance abuse revealed significantly more distress, less adaptive coping styles, and more character pathology than their counterparts. The only significant difference according to UNOS listing was on one measure of family environment. Results support preoperative psychological assessment and intervention on a more extensive level for substance abusers and raise questions for future research. PMID- 11045778 TI - Person reliability of psychiatric patients' responses to a psychopathology inventory. AB - Person-reliability indices can assist clinicians in determining the interpretability of a patient's responses to the Basic Personality Inventory (BPI). Using an initial sample of 65 psychiatric patients, we found that: (1) different person-reliability indices showed modest evidence of psychometric adequacy and tended not to be confounded with general psychopathology; (2) a content consistency index of person reliability was predictably related to other item change variables, whereas within-session profile stability was related to across-session measures of profile stability: and (3) evidence for the ability of person-reliability indices to moderate the validity of clinical criteria was weak. Results provide cautious support for a multidimensional conceptualization of the person reliability construct on the BPI but demand further evaluation of the clinical utility of person reliability indices. PMID- 11045779 TI - Beck depression inventory: exploring its dimensionality in a nonclinical population. AB - The Beck Depression Inventory (BDI) was factor analyzed to examine its potential multidimensionality. Results indicate that the items on the BDI assess two primary dimensions of depression: cognitive-affective and physiological symptoms. Reliabilities for the items comprising each of the factors were acceptable. Between-groups analyses showed that women scored significantly higher than men on both of the factors and on total depression. Within-group analyses showed that levels of cognitive-affective and physiological depression did not differ significantly within the total sample, or for men or women. In future revisions of the BDI, it may be useful to develop scoring systems based on these two dimensions, both to differentiate between types and levels of depression, and to consider implications for prognosis and treatment response. PMID- 11045780 TI - Therapist competence: its temporal course, temporal stability, and determinants in short-term anxiety-provoking psychotherapy. AB - Using hierarchical linear models procedures (Bryk & Raudenbush, 1992), growth curve analyses were performed to examine the temporal course, rate of change over time, and determinants of therapist competence in short-term anxiety-provoking psychotherapy (STAPP) (Sifneos, 1992). Treatments were 20 sessions long, were manualized, and therapists were experienced clinicians receiving manual-guided supervision. Patients (N= 13) had mostly anxiety diagnoses. Results indicated that, on average, therapist competence ratings followed a flat-line course over time. Estimated mean rate of change was close to zero and varied very little across therapists. Also, therapists who, in an initial session, were more competent in performing STAPP also intervened more frequently in a helping and protecting manner. One possible implication of the findings is that one initial evaluation may be an unbiased estimator of a therapist's general level of competence across sessions. PMID- 11045781 TI - The comet assay as a rapid test in biomonitoring occupational exposure to DNA damaging agents and effect of confounding factors. AB - Within the last decade, the comet assay has been used with increasing popularity to investigate the level of DNA damage in terms of strand breaks and alkaline labile sites in biomonitoring studies. The assay is easily performed on WBCs and has been included in a wide range of biomonitoring studies of occupational exposures encompassing styrene, vinyl chloride, 1,3-butadiene, pesticides, hair dyes, antineoplastic agents, organic solvents, sewage and waste materials, wood dust, and ionizing radiation. Eleven of the occupational studies were positive, whereas seven were negative. Notably, the negative studies appeared to have less power than the positive studies. Also, there were poor dose-response relationships in many of the biomonitoring studies. Many factors have been reported to produce effects by the comet assay, e.g., age, air pollution exposure, diet, exercise, gender, infection, residential radon exposure, smoking, and season. Until now, the use of the comet assay has been hampered by the uncertainty of the influence of confounding factors. We argue that none of the confounding factors are unequivocally positive in the majority of the studies. We recommend that age, gender, and smoking status be used as criteria for the selection of populations and that data on exercise, diet, and recent infections be registered before blood sampling. Samples from exposed and unexposed populations should be collected at the same time to avoid seasonal variation. In general, the comet assay is considered a suitable and fast test for DNA-damaging potential in biomonitoring studies. PMID- 11045782 TI - Dietary intake of isothiocyanates: evidence of a joint effect with glutathione S transferase polymorphisms in lung cancer risk. AB - Isothiocyanates (ITCs) are nonnutrient compounds in cruciferous vegetables with anticarcinogenic properties. One proposed mechanism for their protective action is through down-regulation of cytochrome P-450 biotransformation enzyme levels and induction of phase II detoxifying enzymes. Because ITCs also serve as a substrate for GSTs, we evaluated dietary intake of ITCs and GSTM1 and GSTT1 genotype information in a lung cancer case-control study. There were 503 newly diagnosed lung cancer cases (264 men and 239 women) identified from The University of Texas M. D. Anderson Cancer Center and 465 controls (252 men and 213 women) recruited from enrollees in a local managed care organization. Subjects had an in-person interview including a semiquantitative food frequency questionnaire, and blood samples were obtained for genotyping. Cases reported significantly lower ITC intake per day compared with controls (P = 0.009). There was no main effect associated with the GSTM1 null genotype [odds ratio (OR) = 1.09]. However, there was a statistically significant OR of 1.41 associated with the GSTT1 null genotype. On stratified analysis, low ITC intake and the GSTM1 null genotype were associated with increased lung cancer risk in current smokers, with an OR of 2.22 [confidence interval (CI) = 1.20-4.10). For current smokers with the GSTT1 null genotype, the OR with low ITC intake was 3.19 (CI = 1.54 6.62). The comparable OR in the presence of both null genotypes was 5.45 (CI = 1.72-17.22). These effects were not demonstrable for former smokers by GSTM1 genotype, although the risk for low ITC intake and GSTT1 null genotype was 1.79 (CI = 0.95-3.37). Thus, current smokers who are homozygous null for the GST null genotype and who consume less carcinogenic blocking compounds are at higher lung cancer risk. Some of the inconsistencies reported in the role of GST genotypes in lung cancer risk could be due to unexpected confounding from dietary factors. PMID- 11045783 TI - D2 dopamine receptor gene polymorphisms among African-Americans and Mexican Americans: a lung cancer case-control study. AB - Recent research suggests that variant alleles (A1 and B1) of the DRD2 gene play a role in determining smoking status. However, no studies have evaluated these variant alleles in African-Americans and Mexican-Americans. The primary objective of this study, therefore, was to test the hypothesis that ever smokers in these ethnic groups are more likely than never smokers to have the DRD2 alleles associated with tobacco use (A1 and B1). Furthermore, because of a predicted higher prevalence of smokers in a family because of the patterns of inheritance of the genotypes associated with tobacco use, we also anticipated that individuals with these at-risk DRD2 alleles would be more likely to have a family history of smoking-related cancers. Because other inherited genetic variants may interact with smoking on cancer risk, we also hypothesized that this association might differ between cancer patients and control subjects. PCR was used to perform genotyping on peripheral WBC DNA from 140 lung cancer patients (43 Mexican-Americans and 97 African-Americans) and 222 age-, sex-, and ethnicity matched controls (111 Mexican-Americans and 111 African-Americans). A personal family history was obtained from each participant. There were no statistically significant differences in the distribution of the DRD2 genotypes between cases and controls, although the frequency of the B1 genotype significantly differed by ethnicity (P = 0.002 for controls and P = 0.001 for cases). The DRD2 genotypes and smoking status showed a correlation among Mexican-American controls, although not among African-American controls. The cigarette pack-years in control subjects for the two ethnic groups combined were 30.8, 21.9, and 18.6 for the A1A1, A1A2, and A2A2 genotypes and 36.5, 20.8, and 18.5 for the B1B1, B1B2, and B2B2 genotypes, respectively. Similar trends were found for the number of cigarettes smoked per day among control subjects. From the standpoint of polymorphisms, however, there was a borderline significantly increased (3.6 times greater) frequency of smoking-related cancers among the first-degree relatives of case subjects with an A1 allele than among those without an A1 allele. There was also an elevated (1.8 times greater) frequency of smoking-related cancer among first degree relatives of case subjects with a B1 allele compared with patients without a B1 allele, but this finding was not statistically significant. This phenomenon was not observed among control subjects. We noted a trend toward interaction of DRD2 A1 genotypes and case status for increased risk of smoking-related cancer among first-degree relatives. These findings suggest that the variant DRD2 genotypes are associated with a greater likelihood to smoke and a greater smoking intensity, as well as with a familial aggregation of smoking-related cancers. However, a large study is needed to confirm this finding. PMID- 11045784 TI - Molecular changes in second primary lung and breast cancers after therapy for Hodgkin's disease. AB - The risk of lung and breast cancer is significantly increased after therapy for Hodgkin's disease (HD), but there are few data that describe the molecular profiles of these tumors. We investigated the genetic abnormalities in second primary lung (n = 19) and breast cancers (n = 19) that follow therapy for HD ("post-HD cancers") and compared these with changes observed in corresponding tumor types (57 lung and 20 breast cancers) arising in the general population ("sporadic cancers"). DNA obtained from archival tissues was examined using PCR based analyses for loss of heterozygosity and microsatellite alterations (MAs) at several chromosomal regions, TP53 and K-ras gene mutations, and frameshift mutations at minisatellite sequences at the coding regions of several genes (TGF betaRII, IGFIIR, BAX, hMSH6, and hMSH3). The occurrence of loss of heterozygosity at all chromosomal regions taken together and frequencies at most individual areas were similar for the post-HD and sporadic cancers for both lung and breast sites. The overall frequency of MAs in the post-HD tumors was substantially greater (lung, 2.4-fold, P = 0.004; breast, 4.2-fold, P = 0.16) than that in the respective sporadic cancers. No differences in the pattern of TP53 and K-ras mutations were detected between post-HD and sporadic cancers. No mutations were detected at the minisatellite sequences examined. MAs, which reflect widespread genomic instability, occur at greatly increased frequency in post-HD lung and breast cancers. Although the mechanisms underlying the development of increased MAs are unknown, they have been associated with immunosuppression and radiation exposure. Future research should address the role that MAs, as well as other influences, may play in the development of neoplasias that occur after therapy for HD. PMID- 11045785 TI - The p53 codon 72 polymorphism and lung cancer risk. AB - The p53 tumor suppressor gene frequently is mutated in many forms of human carcinomas. A common polymorphism occurs at codon 72 of exon 4, with two alleles encoding either arginine (CGC) or proline (CCC). This p53 polymorphism reportedly is associated with lung cancer susceptibility. However, not all investigations have been consistent, and this hypothesized association remains controversial. We tested the hypothesis that the Pro/Pro genotype is associated with increased lung cancer risk in a large case-control study of lung cancer that included 482 cases and 510 controls from the Massachusetts General Hospital in Boston, Massachusetts. DNA from peripheral blood samples was examined by PCR-RFLP. Pro/Pro homozygotes were found more frequently in adenocarcinomas (cases, 16.4%; controls, 12.0%; P = 0.03). The prevalence of the Pro/Pro homozygous genotype increased in frequency with increasing pack-years of smoking. The combined susceptible genotype homozygous Pro/Pro and heterozygous Arg/Pro was associated with a 1.45-fold higher risk of adenocarcinoma compared with Arg/Arg genotype (95% confidence interval = 1.01-2.06; P = 0.04) after adjustment for relevant variables. Lung adenocarcinoma risk increased with the presence of one or both variant alleles across smoking strata. In addition, at each level of smoking (except nonsmoker and light smoker), the risk associated with smoking was higher for the population with the combined variant (Arg/Pro + Pro/Pro) genotype. The risk for the combined genotype was associated with tobacco exposure status. In conclusion, the codon 72 germ-line polymorphism (Arg/Pro) of the common tumor suppressor gene p53 contributes to heritable susceptibility for smoke-induced lung adenocarcinoma. The modifications by p53 polymorphism and pack-years resulted in an increased risk of the susceptible genotype to lung adenocarcinoma. The p53 gene may modulate the response to environment carcinogens and thereby affect the risk of developing lung adenocarcinoma. PMID- 11045786 TI - Environmental tobacco smoking, mutagen sensitivity, and head and neck squamous cell carcinoma. AB - Although active tobacco smoking has been considered a major risk factor for head and neck cancer, few studies have evaluated environmental tobacco smoke (ETS) and its interaction with mutagen sensitivity on the risk of head and neck cancer. We investigated the relationship between ETS and head and neck cancer in a case control study of 173 previously untreated cases with pathologically confirmed diagnoses of squamous cell carcinoma of the head and neck and 176 cancer-free controls at Memorial Sloan-Kettering Cancer Center between 1992 and 1994. A structured questionnaire was used to collect ETS exposure and other covariates including a history of active tobacco smoking and alcohol use. ETS measures include a history of ETS exposure at home and at workplace. The associations between passive smoking and head and neck cancer were analyzed by Mantel-Haenszel methods and logistic regression models. Additive and multiplicative models were used to evaluate effect modifications between ETS and mutagen sensitivity. The crude odds ratio (OR) for ETS exposure was 2.8 [95% confidence intervals (CI), 1.3-6.0]. Controlling for age, sex, race, education, alcohol consumption, pack years of cigarette smoking, and marijuana use, the risk of squamous cell carcinoma of the head and neck was increased with ETS (adjusted OR, 2.4; 95% CI, 0.9-6.8). Dose-response relationships were observed for the degree of ETS exposure; the adjusted ORs were 2.1 (95% CI, 0.7-6.1) for those with moderate exposure and 3.6 (95% CI, 1.1-11.5) for individuals with heavy exposure (P for trend = 0.025), in comparison with those who never had ETS exposures. These associations and the dose-response relationships were still present when the analysis was restricted to nonactive smoking cases and controls (crude OR, 2.2; 95% CI, 0.6-8.4). Crude odds ratios were 1.8 for those with moderate ETS exposure and 4.3 for individuals with heavy ETS exposure among nonsmoking cases and controls (P for trend = 0.008). More than multiplicative interaction was suggested between passive smoking and mutagen sensitivity. This study suggests that ETS exposure may increase the risk of head and neck cancer with a dose response pattern. Our analysis indicated that passive smoking may interact with mutagen sensitivity and other risk factors to increase the risk of head and neck cancer. Our results need to be interpreted with caution because of potential residual confounding effects of active tobacco smoking and other methodological limitations. Future large-scale studies are warranted to confirm our findings. PMID- 11045787 TI - A meta-analysis of soyfoods and risk of stomach cancer: the problem of potential confounders. AB - It has been suggested that consumption of soyfoods may be associated with a reduction in risk of various cancers, including nonhormonally dependent cancers. The purpose of this meta-analysis was to examine the relationship between fermented and nonfermented soyfoods and risk of stomach cancer. We searched the reference lists of English language publications of diet and stomach cancer studies that were conducted in Asia or among Asians living in the United States or elsewhere between 1966 and 1999. All of the analytic epidemiological studies that obtained individual data on intake of soyfoods and presented risk estimates of the association between intake of soyfoods and risk of stomach cancer were identified and included in this review. Our pooled analysis of 14 studies with data on fermented soyfoods yielded an odds ratio/relative risk of 1.26 (95% confidence interval, 1.11-1.43) in association with high intake of such foods. In contrast, our pooled analysis of 10 studies with data on nonfermented soyfoods found an odds ratio/relative risk of 0.72 (95% confidence interval, 0.63-0.82) in association with high intake of these foods. However, further analyses suggest that fermented and nonfermented soyfoods may be associated with salt and fruit/vegetable intake, respectively; salt and fruit/vegetable intake are directly associated with stomach cancer risk. In almost all of the studies we reviewed, the possible confounding role of salt, fruit/vegetable, and other dietary factors had not been considered in the soyfood analyses. In conclusion, the role of soyfoods in the etiology of stomach cancer cannot be determined with confidence until the roles of potential confounders, including salt, fruit/vegetables, and other dietary factors, are more adequately adjusted for. PMID- 11045788 TI - Plasma 1,25-dihydroxy- and 25-hydroxyvitamin D and adenomatous polyps of the distal colorectum. AB - 1,25-dihydroxyvitamin D [1,25(OH)2D] inhibits proliferation and promotes differentiation of human colon cancer cell lines. Epidemiological findings, although not entirely consistent, suggest an inverse relationship between vitamin D intake and colorectal cancer and adenoma, colorectal cancer precursor lesions. We evaluated the relationship of plasma 1,25(OH)2D and 25-hydroxyvitamin D [25(OH)D] with distal colorectal adenoma among 326 matched case and control pairs (nested in the prospective Nurses' Health Study), who provided blood in 1989-1990 and who underwent endoscopy in 1989-1996. Plasma vitamin D metabolite concentrations were determined blindly by RIA. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated from multiple conditional logistic regression models. Mean plasma 1,25(OH)2D and 25(OH)D levels did not significantly differ (P = 0.3 and 0.7, respectively) between cases (31.6 +/- 8.4 pg/ml and 26.4 +/- 10.6 ng/ml, respectively) and controls (32.2 +/- 8.6 pg/ml and 26.8 +/-10.2 ng/ml, respectively). However, women whose plasma 1,25(OH)2D concentration was below 26.0 pg/ml (a level typically considered to be below normal) were at increased risk of distal colorectal adenoma (OR, 1.58; 95% CI, 1.03-2.40). Compared with the lowest 1,25(OH)2D quartile, women in the second (OR, 0.64; 95% CI, 0.41-1.02), third (OR, 0.80; 95% CI, 0.50-1.30), or upper (OR, 0.71; 95% CI, 0.43-1.15) quartiles were at a statistically nonsignificant lower risk of adenoma. The relationship was stronger for large/villous adenoma and among those with consistent vitamin D intake over the 10 years prior to blood draw. Compared with women in the lowest quartile, for plasma 25(OH)D, women in the second (OR, 0.64; 95% CI, 0.41-1.00) and third (OR, 0.58; 95% CI, 0.36-0.95) quartiles were at a statistically significantly lower risk of distal colorectal adenoma, but there was no difference in risk in the top quartile (OR, 1.04; 95% CI, 0.66-1.66). We conclude that women who have low levels of circulating 1,25(OH)2D may be at higher risk of distal colorectal adenomas, but additional study is warranted. PMID- 11045789 TI - Tumor necrosis factor a-11 and DR15-DQ6 (B*0602) haplotype increase the risk for cervical intraepithelial neoplasia in human papillomavirus 16 seropositive women in Northern Sweden. AB - HLA genes have been shown to be associated with cervical intraepithelial neoplasia (CIN), a precursor of cervical cancer. The human papillomaviruses (HPV) types 16 and 18 are the major environmental cause of this disease. Because the immune system plays an important role in the control of HPV infection, the association of polymorphic HLA could lead to a different immune response to control the development of cervical cancer. The aim of this study was to analyze the association between CIN and a microsatellite polymorphism of tumor necrosis factor (TNFa) taking HPV exposure and CIN-associated HLA haplotypes into account. In a nested case-control study in northern Sweden, 64 patients and 147 controls matched for age and sex and derived from the same population-based cohort were typed for TNFA, HLA-DR, and DQ and assayed for antibodies to HPV types 16 and 18. TNFa polymorphism was not associated with CIN per se. However, there was a significant increase in the frequency of TNFa-11 among HPV16-positive and HLA DR15-DQ6 (B*0602) patients compared with HPV16- and HLA-DQ6-negative patients (odds ratios, 5.4 and 9.3, respectively). The relative risk for CIN conferred by the combination of TNFa-11, HLA-DQ6, and HPV 16 positivity was 15. Our study suggests that the TNFa-11 allele is associated with HPV16 infection and associated with CIN in combination with HLA-DQ6 but not by itself. PMID- 11045790 TI - Randomized trial of fenretinide in superficial bladder cancer using DNA flow cytometry as an intermediate end point. AB - Retinoids have shown a potential activity in preventing tumor recurrence in superficial bladder cancer. We assessed the activity of the synthetic retinoid fenretinide in superficial bladder cancer using DNA flow cytometry and conventional cytology as surrogate biomarkers. A total of 99 subjects with resected superficial bladder cancer (pTa, pT1) were randomized to either fenretinide (200 mg day p.o. for 24 months) or no intervention. Cystoscopy and bladder washing for DNA flow cytometry end points (proportion of DNA aneuploid histograms, hyperdiploid fraction, and percentage of apoptotic cells) and proportion of abnormal cytological examinations were repeated every 4 months for up to 36 months. The primary study end point was the proportion of DNA aneuploid histograms after 12 months. This figure was 48.9% in the fenretinide arm and 41.9% in the control arm (odds ratio, 1.16; 95% confidence interval, 0.44-3.07). There was no difference in any other response biomarker between the two groups up to 36 months, nor was any biomarker able to predict recurrence risk. Recurrence free survival was comparable between the arms (27 events in the fenretinide arm versus 21 in the control arm; P = 0.36). Twelve subjects in the fenretinide arm complained of diminished dark adaptability, and nine subjects in the fenretinide arm versus one control subject had mild dermatological alterations. We conclude that fenretinide showed a lack of effect on the DNA content distribution and the morphology of urothelial cells obtained in serial bladder washings. Recurrence free survival was comparable between groups. Because our data are hampered by the lack of predictivity of the selected biomarkers, additional studies are necessary to assess the activity of fenretinide in preventing bladder cancer. PMID- 11045791 TI - The association between glutathione S-transferase M1 genotype and polycyclic aromatic hydrocarbon-DNA adducts in breast tissue. AB - A major goal in molecular epidemiology is to identify preventable environmental risk factors and susceptible subpopulations. In a hospital-based molecular epidemiological case-control study of breast cancer, we investigated the relationship between DNA damage from exposure to polycyclic aromatic hydrocarbons (PAHs) and susceptibility attributable to inherited deletion of the xenobiotic detoxifying gene, glutathione S-transferase M1 (GSTM1). Prior to breast surgery, women (n = 227) were enrolled and interviewed and donated a blood sample. PAH-DNA adduct levels were measured by immunohistochemistry in breast tissue samples retrieved from pathology blocks, and GSTM1 genotype was determined by PCR using WBC DNA. The GSTM1 analysis included 95 cases and 87 benign breast disease controls. GSTM1 genotype was not associated with breast cancer case-control status (odds ratio = 0.73; 95% confidence interval, 0.37-1.44). However, the GSTM1 null genotype predicted PAH-DNA adduct levels in malignant (beta = 0.407; P = 0.003) and nonmalignant (beta = 0.243; P = 0.05) breast tissue from cases. This relationship was not seen in tissue from controls (beta = 0.095; P = 0.341). When tissue from controls was compared with tumor tissue from cases, there was a significant case-control difference in PAH-DNA adduct levels among women who were GSTM1 null. There was no such case-control difference among women who were homozygous or heterozygous for GSTM1. There was an interaction between GSTM1 and case-control status on adduct levels in breast tissue (P = 0.002). The results suggest that genetic susceptibility to the formation of PAH-DNA adducts in breast tissue may play a role in breast cancer development. PMID- 11045792 TI - Novel translational model for breast cancer chemoprevention study: accrual to a presurgical intervention with tamoxifen and N-[4-hydroxyphenyl] retinamide. AB - Surrogate end point biomarkers for risk assessment and efficacy of potential chemopreventive agents are needed to improve the efficiency and reduce the cost of chemoprevention trials. It is imperative to develop the best clinical breast model for translational surrogate end point biomarker studies, especially with respect to accrual feasibility. We have initiated a prospective study to develop biomarkers for tamoxifen and N-[4-hydroxyphenyl] retinamide by administering either a placebo or both drugs for 2-4 weeks to women with ductal carcinoma in situ or early invasive cancers in the interval between the initial diagnostic core biopsy and definitive surgery. The principle end point is pretreatment versus posttreatment tumor levels of Ki-67; a number of other exploratory markers will also be examined. The planned target sample size is 100 patients. Between February 1997 and February 2000, 4514 women who had either an abnormal mammogram or a diagnosed breast cancer were screened for the study. Of these 4514 screened patients, 52 (1%) were registered on the study. Major factors of nonparticipation in the remaining 4462 women were as follows: (a) no evidence of malignancy (2081 patients; 46%); (b) ineligible per protocol criteria (575 patients; 13%); (c) preoperative chemotherapy/tamoxifen (520 patients; 11%); (d) surgery scheduling conflict (360 patients; 8%); (e) outside needle biopsy (221 patients; 5%); (f) no residual disease after excisional biopsy (345 patients; 8%); and (g) second opinion only (123 patients; 3%). Other nonparticipation factors included fine needle aspiration only, refusal, tumor size > 2 cm, and estrogen replacement therapy (35 patients each; 2% each). The protocol was amended in midstudy to allow outside needle biopsy, tumor > 2 cm, and estrogen replacement therapy. Accrual to biomarker (nontherapeutic) protocols with delay in definitive cancer surgery is challenging but feasible. Although some accrual problems remain, we have nonetheless succeeded in recruiting 50% of our target sample size in a 3 year period. PMID- 11045793 TI - Low-energy reporting in women at risk for breast cancer recurrence. Women's Healthy Eating and Living Group. AB - This study examined the extent of low-energy reporting and its relationship with demographic and lifestyle factors in women previously treated for breast cancer. This study used data from a large multisite clinical trial testing the efficacy of a dietary intervention to reduce risk for breast cancer recurrence (Women's Healthy Eating and Living Study). Using the Schofield equation to estimate energy needs and four 24-h dietary recalls to estimate energy intakes, we identified women who reported lower than expected energy intakes using criteria developed by G. R. Goldberg et al. (Eur. J. Clin. Nutr., 45: 569-581, 1991). We examined data from 1137 women diagnosed with stage I, stage II, or stage IIIA primary, operable breast cancer. Women were 18-70 years of age at diagnosis and were enrolled in the Women's Healthy Eating and Living Study between August 19, 1995, and April 1, 1998, within 4 years after diagnosis. The Goldberg criteria classified about one quarter (25.6%) as low-energy reporters (LERs) and 10.8% as very LERs. Women who had a body mass index >30 were almost twice (odds ratio, 1.95) as likely to be LERs. Women with a history of weight gain or weight fluctuations were one and a half times as likely (odds ratio, 1.55) to be LERs as those who were weight stable or weight losers. Age, ethnicity, alcohol intake, supplement use, and exercise level were also related to LER. Characteristics (such as body mass index, age, ethnicity, and weight history) that are associated with low-energy reporting in this group of cancer survivors are similar to those observed in other populations and might affect observed diet and breast cancer associations in epidemiological studies. PMID- 11045794 TI - A prospective study of the effect of alcohol consumption and ADH3 genotype on plasma steroid hormone levels and breast cancer risk. AB - One suggested mechanism underlying the positive association between alcohol consumption and breast cancer risk is an influence of alcohol on steroid hormone levels. A polymorphism in alcohol dehydrogenase type 3 (ADH3) affects the kinetics of alcohol oxidation and thereby could influence the effect of alcohol consumption on hormone levels. We investigated the ADH3 polymorphism, alcohol intake, and risk of breast cancer in a nested case-control study. Among women in the Nurses' Health Study who gave a blood sample in 1989-1990, 465 incident breast cancer cases were diagnosed before June 1994 and were matched to 621 controls. Using conditional logistic regression, we calculated relative risks and confidence intervals to assess breast cancer risk for ADH3 genotype. Among postmenopausal controls not using hormones at time of blood collection, partial Pearson correlation coefficients were calculated to assess the association between alcohol intake and plasma hormone levels according to ADH3 genotype. No association was observed between ADH3 genotype and overall breast cancer risk (relative risk = 0.9 for slow oxidizers compared with fast; 95% confidence interval = 0.6-1.3). Among postmenopausal women, ADH3 genotype did not modify the weak association observed between alcohol intake and breast cancer risk (P for interaction = 0.45). Statistically significant trends in the relationship between alcohol consumption and hormone level dependent on oxidative capacity (ADH3 genotype) were observed for dehydroepiandrosterone sulfate and sex hormone binding globulin (P < 0.05). These data suggest that the ADH3 polymorphism modestly influences the response of some plasma hormones to alcohol consumption but is not independently associated with breast cancer risk and does not modify the association between alcohol and breast cancer risk. PMID- 11045795 TI - Ovarian cancer tumor marker behavior in asymptomatic healthy women: implications for screening. AB - Ovarian cancer screening protocols generally have been limited by inadequate recognition of the normal behavior of tumor markers in women at risk of ovarian cancer. We have characterized the behavior of five serum tumor markers in a large cohort of healthy women and examined the implications for screening. Serial measurements of CA125, HER-2/neu, urinary gonadotropin peptide, lipid-associated sialic acid, and Dianon marker 70/K were obtained during 6 years of follow-up of 1257 healthy women at high risk of ovarian cancer. We analyzed individual specific tumor marker behavior and explored methods that can exploit this information to develop individual-specific screening rules. These five tumor markers behaved approximately independently. Substantial heterogeneity was observed among women in the behavior of each tumor marker, particularly CA125. Intraclass correlation (ICC), or the proportion of total variability that occurs between individuals, was approximately 0.6 for log-transformed CA125 and urinary gonadotropin peptide, and less than 0.4 for the other markers. This degree of tumor marker heterogeneity among healthy women, and the relative independence of these markers, has important implications for screening and diagnostic tests. Independence of markers results in the clinically useful fact that the combined false positive rate from screening with multiple markers may be estimated by the sum of individual false positive rates. Heterogeneity of tumor marker patterns in healthy women implies that a fixed screening cutoff level is suboptimal to a degree that depends strongly on ICC. Using information on longitudinal measurements and ICC, individual-specific screening rules may be developed with the potential to improve early detection of ovarian cancer. PMID- 11045796 TI - Flaxseed influences urinary lignan excretion in a dose-dependent manner in postmenopausal women. AB - Dietary estrogens, such as lignans, are similar in structure to endogenous sex steroid hormones and may act in vivo to alter hormone metabolism and subsequent cancer risk. The objective of this study was to examine the effect of dietary intake of a lignan-rich plant food (flaxseed) on urinary lignan excretion in postmenopausal women. This randomized, cross-over trial consisted of three 7-week feeding periods during which 31 healthy postmenopausal women, ages 52-82 years, consumed their habitual diets plus 0, 5, or 10 grams of ground flaxseed per day. Urine samples collected for 2 consecutive days during the last week of each feeding period were analyzed for lignan content (enterodiol, enterolactone, and matairesinol) by isotope dilution gas chromatography/mass spectrometry. Compared with the 0-gram flaxseed diet, consumption of 5 or 10 grams of flaxseed significantly increased excretion of enterodiol by 1,009 and 2,867 nmol/day, respectively; significantly increased excretion of enterolactone by 21,242 and 52,826 nmol/day, respectively; and significantly increased excretion of total lignans (enterodiol + enterolactone + matairesinol) by 24,333 and 60,640 nmol/day, respectively. Excretion of matairesinol was not significantly altered by flaxseed consumption. Consumption of flax, a significant source of dietary estrogens, in addition to their habitual diets increased excretion of enterodiol and enterolactone, but not matairesinol, in a dose-dependent manner in this group of postmenopausal women. Urinary excretion of lignan metabolites is a dose dependent biomarker of flaxseed intake within the context of a habitual diet. PMID- 11045797 TI - CYP1A1, GSTM1, and GSTP1 genetic polymorphisms and urinary 1-hydroxypyrene excretion in non-occupationally exposed individuals. AB - The CYP1A1 and glutathione S-transferase enzymes (e.g., GSTM1 and GSTP1) are involved in the activation and conjugation of polycyclic aromatic hydrocarbons (PAHs), respectively, and are controlled by genes that are polymorphic. The CYP1A1*2 allelic variant has been associated with elevated urinary 1 hydroxypyrene (1-OHP), a proposed marker for internal dose of activated PAHs, in coke-oven workers. We investigated whether this association could be observed at low exposure levels, such as those experienced by the general population. We conducted a cross-sectional study among 188 individuals (106 Japanese, 60 Caucasians, and 22 Hawaiians) who were selected as controls in a population-based case-control study and provided lifestyle information, a 12-h urine specimen, and a blood sample. 1-OHP was analyzed by high-performance liquid chromatography after enzymatic hydrolysis. Lymphocyte DNA was used for PCR-based genotyping. Smokers excreted twice as much 1-OHP (geometric mean, 0.51 nmol/12 h) as nonsmokers (geometric mean, 0.27 nmol/12 h; P = 0.006). Overall and among nonsmokers, 1-OHP urinary levels did not differ by CYP1A1, GSTM1, or GSTP1 genotypes. However, after adjusting for age, ethnicity, and number of cigarettes per day, smokers with at least one CYP1A1*2 variant allele excreted 2.0-fold more 1-OHP than smokers with the wild-type genotype (P = 0.02). Similar results were obtained for the CYP1A1*3 variant allele. The present data add to the growing evidence suggesting that individuals with the (linked) CYP1A1*2 or *3 variant alleles have a greater capacity to activate PAHs from tobacco smoke and occupational exposure and, as a result, are at greater risk for PAH-related cancers, especially certain respiratory cancers. PMID- 11045798 TI - Disposable versus reusable biopsy forceps for colorectal epithelial cell proliferation in humans. AB - The performance of various measures of rectal mucosal proliferation has been evaluated in the literature, but the performance of the forceps used to obtain the tissue has received little attention. We used data from two large studies of proliferation at a single institution to compare reusable and disposable endoscopic forceps. Endoscopic pinch biopsies were taken 10 cm from the anal verge using either reusable or disposable, oval-cupped, sheathed forceps. The specimens were fixed, embedded, and sectioned, taking care to orient the specimens longitudinally. Five sections were placed on each slide. We determined how many slides did not contain eight scorable crypts (inadequate) and how many sections were necessary to identify eight complete crypts. There were 395 subjects who had biopsies taken with reusable forceps and 185 subjects who had biopsies taken with disposable forceps. The specimens were inadequate in 27.6% of the reusable forceps specimens versus 2.7% of the disposable forceps (P < 0.0001). The mean number of tissue sections necessary to identify eight scorable crypts for the reusable forceps was 3.82 (SD, 0.87) compared with 3.17 (SD, 0.83) for disposable forceps (P = 0.0001). The specimens taken with the disposable forceps were better, probably because the forceps were sharper. We believe that the better quality of the specimens and the sterility justify the higher cost of disposable forceps. We would urge investigators in proliferation studies to evaluate the biopsy equipment as carefully as they evaluate other aspects of their methods. PMID- 11045799 TI - Participation in a sigmoidoscopic colorectal cancer screening program: a pilot study. AB - At present, very little is known about the determinants of endoscopic screening participation. This study presents an analysis of the psychosocial associations of participation and nonparticipation in a sigmoidoscopic colorectal cancer screening program. The present pilot study was executed among members of a Dutch target group, ages 50-60 years, who visited an internal medicine outpatient clinic. Individuals who were asked to participate in the program (n = 200) received general information with regard to the screening procedure. The participation rate was 45%. Persons who participated in the screening program as well as those who wanted to participate in the study but did not want to participate in the screening program were asked to fill out a questionnaire. Self efficacy, i.e., the individual's perception of the difficulty of participating in the screening program, appeared to be the most important association of participation. Furthermore, response efficacy, i.e., the individual's beliefs about the outcome of participation, and social support proved to be concepts that were associated with participation. PMID- 11045800 TI - Strategies for chemical reaction searching in SciFinder AB - The bibliographic, chemical structure, and chemical reaction databases produced by Chemical Abstracts Service allow a number of possibilities for chemical reaction searching. While these same databases may be searched through the STN network, many end-users find the intuitive software interface SciFinder simpler, but there still are issues to address. Searching may be performed through keywords, chemical structures, or chemical reactions, and the answers may vary with respect to precision and comprehension. Often combinations of search options may be needed to best solve the problem. Retrosynthetic analyses are easily performed in the chemical reaction database and can give unique insights into synthetic alternatives. PMID- 11045801 TI - A revisited auditing of the analytical abstracts database AB - This paper is a follow-up of a previous one dealing with the "Image of Analytical Chemistry as Reflected in the Analytical Abstracts Database: Journal Coverage, Concentration and Dispersion of the Analytical Literature" (J. Chem. Inf. Comput. Sci. 1993, 33, 164-173). It deals with revisiting these topics. The results have shown that the database has substantially improved its coverage by editorial reorganizations in 1994. The only open problem which has been revealed is a somewhat excessive emphasis given to the coverage of the journals on the lower tail of the journal distribution. The suggestion is made to reduce this emphasis in favor of an even more complete coverage of some "titled" analytical journals. PMID- 11045802 TI - Multivariate analysis of near-infrared spectra using the G-programming language AB - "Real-time" chemometrics as envisioned by the union of instrument control, data acquisition, and chemometric analysis with a single software platform can provide substantial benefits to manufacturing concerns that require process control. Some of these benefits include faster generation of information and improved quality control. This paper describes a series of chemometric routines written in LabVIEW and demonstrates their use in predicting six properties of diesel fuel. In particular, near-infrared spectral data were used to predict the boiling point at 50% recovery, cetane number, density, freezing temperature, total aromatics, and viscosity for a series of diesel fuels. PMID- 11045803 TI - Characteristic monomials with chirality fittingness for combinatorial enumeration of isomers with chiral and achiral ligands AB - A new method of combinatorial enumeration based on characteristic monomials with chirality fittingness (CM-CFs) has been proposed in order to enumerate isomers with chiral ligands as well as with achiral ones. The CM-CFs have been defined as monomials that consist of three kinds of dummy variables in light of the subduction of the Q-conjugacy representations for chiral and achiral cyclic groups. A procedure of calculating CM-CFs for cyclic groups and finite groups has been discribed so as to tabulate them as CM-CF tables. Then the CM-CF method has been applied to the enumeration of isomers with achiral ligands as well as chiral ones. PMID- 11045804 TI - QSAR treatment of electronic substituent effects using frontier orbital theory and topological parameters AB - A methodology for the estimation of Hammett substituent constants from computational-based descriptors utilizing quantitative structure activity/property relationships (QSAR/QSPR) formalism is presented. Electronic descriptors derived from quantum chemical calculations and molecular topology were used to generate computational-based analogues of empirical Hammett substituent constants from statistical analysis. Global quantum chemical reaction indices were drawn from frontier orbital theory and density functional theory and formulated from AM1-based calculations. A localized index based on the electrotopological state index was used to encode information on individual group properties. From a training set consisting of 150 meta and para-substituted benzoic acids, statistical analysis of computational-based descriptors as a function of empirical substituent constants yielded a five-parameter QSAR/QSPR model which generates computational-based constants exhibiting a strong correlation with empirical values (r2 = 0.958). Both internal (PRESS) and external (independent testing set of benzoic acids) validation procedures suggest that the electronic effects QSAR/QSPR model derived in this work from computational-based parameters is a statistically viable paradigm. Both predicted and empirical constants were used in Hammett-type validation analyses as functions of chemical, biological, and spectroscopic data for thirty structurally diverse meta and parasubstituted aromatic testing sets. Statistical measures of ensuing correlations were examined and compared, and the empirical and predicted results were of similar quality. Validation results reveal that a large number of computational-based substituent constants can be accurately estimated from semiempirical AM1 frontier orbital energies and electronic structure information obtained directly from substituted benzoic acids without the aid of empirical parametrization. PMID- 11045805 TI - Identification of groupings of graph theoretical molecular descriptors using a hybrid cluster analysis approach. AB - There is an abundance of structural molecular descriptors of various forms that have been proposed and tested over the years. Very often different descriptors represent, more or less, the same aspects of molecular structures and, thus, they have diminished discriminating power for the identification of different structural features that might contribute to the molecular property, or activity of interest. Therefore, it is essential that noncorrelated descriptors be employed to ensure the wider and the less inflated possible coverage of the chemical space. The most usual approach for reducing the number of descriptors and employing noncorrelated (or orthogonal) descriptors involves principal component analysis (PCA) or other factor analytical techniques. In this work we present an approach for determining relationships (groupings) among 240 graph theoretical descriptors, as a means for selecting nonredundant ones, based on the application of cluster analysis (CA). To remove inherent biases and particularities of different CA algorithms, several clustering solutions, using these algorithms, were "hybridized" to obtain a reliable and confident overall solution concerning how the interrelationships within the data are structured. The calculated correlation coefficients between descriptors were used as a reference for a discussion on the different CA methods employed, and the resulted clusters of descriptors were statistically analyzed for deriving the intercorrelations between the different operators, weighting schemes and matrices used for the computation of these descriptors. PMID- 11045806 TI - A compact form of the adjacency matrix AB - It has been shown that the adjacency matrix can be transformed into a row vector and then into a single number. This number can again be decoded to recover the row vector, and this in turn can be decoded to restore the original adjacency matrix. A special, rather efficient coding scheme was devised for acyclic structures. PMID- 11045807 TI - Stability of complexes of aromatic amides with bromide anion: quantitative structure--property relationships AB - Most of the theoretical studies published to-date on the structural and electronic properties of supramolecules have been devoted to the neutral or cationic complexes, while little is known about anionic systems. A detailed theoretical study of the interaction between simple aromatic amides and the bromide anion has recently been published (Cajan, M.; Stibor, I.; Koca, J. J. Phys. Chem. A 1999, 103, 3778). The present work focuses on the structural and physicochemical parameters of simple aromatic amides related to their ability to form the 1:1 complex with a bromide anion. A quantitative structure-property relationships (QSPR) model for the prediction of association constants is proposed. The model based on 22 complexes and nine molecular descriptors explained 96% (84% cross-validated) of the variance in association constants. The descriptors employed in this model included parameters for the characterization of conformational behavior and the 3D structure of amide molecules, distribution of electron density on the amidic functional group, and parameters for substitution on aromatic units. The quantitative structure-property relationship approach predicted the association constants with comparable quality, but significantly lower computational demand, than molecular modeling or standard quantum chemistry calculations. PMID- 11045808 TI - Topological organic chemistry. 13. Transformation of graph adjacency matrixes to distance matrixes AB - A sequence of simple arithmetic operations is described that generates distance matrixes from the adjacency matrixes of graphs. PMID- 11045809 TI - Unsupervised forward selection: a method for eliminating redundant variables. AB - An unsupervised learning method is proposed for variable selection and its performance assessed using three typical QSAR data sets. The aims of this procedure are to generate a subset of descriptors from any given data set in which the resultant variables are relevant, redundancy is eliminated, and multicollinearity is reduced. Continuum regression, an algorithm encompassing ordinary least squares regression, regression on principal components, and partial least squares regression, was used to construct models from the selected variables. The variable selection routine is shown to produce simple, robust, and easily interpreted models for the chosen data sets. PMID- 11045810 TI - Fast determination of 13C NMR chemical shifts using artificial neural networks. AB - Nine different artificial neural networks were trained with the spherically encoded chemical environments of more than 500000 carbon atoms to predict their 13C NMR chemical shifts. Based on these results the PC-program "C_shift" was developed which allows the calculation of the 13C NMR spectra of any proposed molecular structure consisting of the covalently bonded elements C, H, N, O, P, S and the halogens. Results were obtained with a mean deviation as low as 1.8 ppm; this accuracy is equivalent to a determination on the basis of a large database but, in a time as short as known from increment calculations, was demonstrated exemplary using the natural agent epothilone A. The artificial neural networks allow simultaneously a precise and fast prediction of a large number of 13C NMR spectra, as needed for high throughput NMR and screening of a substance or spectra libraries. PMID- 11045812 TI - Molecular electronic density fitting using elementary Jacobi rotations under atomic shell approximation AB - Fitted electron density functions constitute an important step in quantum similarity studies. This fact not only is presented in the published papers concerning quantum similarity measures (QSM), but also can be associated with the success of the developed fitting algorithms. As has been demonstrated in previous work, electronic density can be accurately fitted using the atomic shell approximation (ASA). This methodology expresses electron density functions as a linear combination of spherical functions, with the constraint that expansion coefficients must be positive definite, to preserve the statistical meaning of the density function as a probability distribution. Recently, an algorithm based on the elementary Jacobi rotations (EJR) technique was proven as an efficient electron density fitting procedure. In the preceding studies, the EJR algorithm was employed to fit atomic density functions, and subsequently molecular electron density was built in a promolecular way as a simple sum of atomic densities. Following previously established computational developments, in this paper the fitting methodology is applied to molecular systems. Although the promolecular approach is sufficiently accurate for quantum QSPR studies, some molecular properties, such as electrostatic potentials, cannot be described using such a level of approximation. The purpose of the present contribution is to demonstrate that using the promolecular ASA density function as the starting point, it is possible to fit ASA-type functions easily to the ab initio molecular electron density. A comparative study of promolecular and molecular ASA density functions for a large set of molecules using a fitted 6-311G atomic basis set is presented, and some application examples are also discussed. PMID- 11045811 TI - Drug-like index: a new approach to measure drug-like compounds and their diversity. AB - Combinatorial organic synthesis (combinatorial chemistry or CC) and ultrahigh throughput screening (UHTS) are speeding up drug discovery by increasing capacity for making and screening large numbers of compounds. However, a key problem is to select the smaller set of "representative" compounds from a virtual library to make or screen. Our approach is to select drug-like as well as structurally diverse compounds. The compounds, which are not very drug-like, are less taken into account or excluded even if they contribute to the diversity of the collection. Hence, the first step in the compound selection is to rank compounds in drug-like "degree". To quantify the drug-like "degree", drug-like index (DLI) is introduced in this paper. A compound's DLI is calculated based upon the knowledge derived from known drugs selected from Comprehensive Medicinal Chemistry (CMC) database. The paper describes the way of this knowledge base is formed and the procedure for selecting drug-like compounds. PMID- 11045813 TI - Database of C-glycosylporphyrins in Web fashion. AB - The aim of this work was to organize chemical data in a client-server environment using Database Management System and Web fashion for the client interface. To solve this ancient problem (for us) merging text data, reaction schemes, tridimensional structures, and NMR, CD, and UV spectra images, we have based our implementation on a few fundamental points: no cost for the user, availability of data via the Internet, standard and freeware software, and a Web browser for the database inquiry. These functions are delivered in a platform-independent manner via the Internet and are used by computational experts and nonexperts alike. C Glycosylporphyrins is the class of compounds chosen to test our applications. These results can be exportable for many other classes of chemical compounds. PMID- 11045814 TI - The permanental polynomial. AB - This study identifies properties and uses of the permanental polynomial of adjacency matrixes of unweighted chemical graphs. Coefficients and zeroes of the polynomial for several representative structures are provided, and their properties are discussed. A computer program for calculating the permanental polynomial from the adjacency matrix is also described. PMID- 11045815 TI - Permanental polynomials of the smaller fullerenes. AB - Using a general computer code developed previously, permanental polynomials were computed for all fullerenes C< or =36. Mathematical properties of the coefficients and zeroes were investigated. For a given isomer series of constant n, the n/2 independent zeroes appear to consist of a set of 10 that are nearly constant within the series and a set of n/2-10 that differ greatly with structure. PMID- 11045816 TI - Classification of some active compounds and their inactive analogues using two three-dimensional molecular descriptors derived from computation of three dimensional convex hulls for structures theoretically generated for them. AB - Two three-dimensional (3D) molecular descriptors are used to classify 73 protease inhibitors against the human immunodeficiency virus type 1 (HIV-1). X-ray structures of these HIV-1 protease bound inhibitors are used as templates to generate the most probable bioactive conformations of the inhibitors. A convex hull computation algorithm is applied to each structure generated. The frequency of atoms lying on the vertexes of each hull is counted. Vertexes of the same atomic charge state are then gathered together as a set of commonly exposed groups for all the structures generated. The first 3D descriptor is computed as the maximum molecular path length among any three distinct commonly exposed groups, while the second 3D one is computed as the maximum molecular path length among any three atoms of nonconvex hull vertexes. We find that the 73 HIV-1 protease inhibitors can be classified by the first 3D descriptor into two groups, which agrees with the result of visual classification using the activity data as a criterion for these compounds. The classification scheme is then used to classify a database of 427 active trypsin inhibitors and their inactive analogues. The structures of these compounds are generated theoretically from steps of energy minimization and molecular dynamics. Classification for all these compounds is performed using the SYBYL hierarchical clustering method on the first 3D descriptor and then the second 3D one computed. It is found that some inactive analogues are completely separated from the active inhibitors at the first stage of classification using the first 3D descriptor. Most of the highly active inhibitors are classified into a cluster at the second stage of classification using the second 3D descriptor. Finally, most of these highly active inhibitors are separated from all the accompanying inactive analogues in the cluster through a structural alignment process using a set of commonly exposed groups determined for them. PMID- 11045817 TI - A new method for estimation of homolytic C-H bond dissociation enthalpies AB - In this work we have quantitatively analyzed substituent effects on the homolytic bond dissociation enthalpy of 79 different compounds using a method based on discrete distance dependent atomic contributions to a molecular property. The resulting empirical relationship can be used to predict C-H bond dissociation enthalpies (within +/-10 kJ mol(-1)) for molecules where resonance contributions and captodative stabilization are insignificant. For species where captodative stabilization of the corresponding C-centered radical is possible, the method clearly overestimates the C-H bond dissociation enthalpy. PMID- 11045818 TI - Evaluation of descriptors and mini-fingerprints for the identification of molecules with similar activity. AB - Combinations of 65 preferred 1D/2D molecular descriptors and 143 single structural keys were evaluated for their performance in compound classification focused on biological activity. The analysis was based on principal component analysis of descriptor combinations and facilitated by use of a genetic algorithm and different scoring functions. In these calculations, several descriptor combinations with greater than 95% prediction accuracy were identified. A set of 40 preferred structural keys was incorporated into a small binary fingerprint designed to search databases for compounds with biological activity similar to query molecules. The performance of mini-fingerprints was tested by systematic similarity search calculations in a database consisting of compounds belonging to seven biological activity classes, which had not been used to select effective descriptors. In these blind test calculations, mini-fingerprints correctly identified approximately 54% of compounds sharing similar biological activity and with 1% false positives. Thus, although the design of mini-fingerprints is conceptually simple, they perform well in activity-oriented similarity searching. PMID- 11045819 TI - On 3-D graphical representation of DNA primary sequences and their numerical characterization. AB - In this article we (1) outline the construction of a 3-D "graphical" representation of DNA primary sequences, illustrated on a portion of the human beta globin gene; (2) describe a particular scheme that transforms the above 3-D spatial representation of DNA into a numerical matrix representation; (3) illustrate construction of matrix invariants for DNA sequences; and (4) suggest a data reduction based on statistical analysis of matrix invariants generated for DNA. Each of the four contributions represents a novel development that we hope will facilitate comparative studies of DNA and open new directions for representation and characterization of DNA primary sequences. PMID- 11045820 TI - Distinguishing between natural products and synthetic molecules by descriptor Shannon entropy analysis and binary QSAR calculations. AB - Molecular descriptors were identified by Shannon entropy analysis that correctly distinguished, in binary QSAR calculations, between naturally occurring molecules and synthetic compounds. The Shannon entropy concept was first used in digital communication theory and has only very recently been applied to descriptor analysis. Binary QSAR methodology was originally developed to correlate structural features and properties of compounds with a binary formulation of biological activity (i.e., active or inactive) and has here been adapted to correlate molecular features with chemical source (i.e., natural or synthetic). We have identified a number of molecular descriptors with significantly different Shannon entropy and/or "entropic separation" in natural and synthetic compound databases. Different combinations of such descriptors and variably distributed structural keys were applied to learning sets consisting of natural and synthetic molecules and used to derive predictive binary QSAR models. These models were then applied to predict the source of compounds in different test sets consisting of randomly collected natural and synthetic molecules, or, alternatively, sets of natural and synthetic molecules with specific biological activities. On average, greater than 80% prediction accuracy was achieved with our best models. For the test case consisting of molecules with specific activities, greater than 90% accuracy was achieved. From our analysis, some chemical features were identified that systematically differ in many naturally occurring versus synthetic molecules. PMID- 11045821 TI - Improved quantitative structure property relationships for the prediction of dielectric constants for a set of diverse compounds by subsetting of the data set AB - In a recent publication we explored the development of quantitative structure property relationships for the calculation of dielectric constants, which resulted in a general model for a wide range of compounds. Our current work explores the division of the set of compounds into eight more homogeneous subsets for which local models are developed. The full data set consists of 454 compounds with dielectric constants ranging from 1 to 40. A pool of up to 16 molecular descriptors is calculated for each of the eight data sets. The descriptors include dipole moment, polarizability, counts of elemental types or functional groups, charged partial surface area, and molecular connectivity. All possible 4 16 descriptor models are calculated for each of the eight data sets, and the best models are selected and compared to the results obtained from the best general model for all 454 compounds. Neural networks using the Broyden-Fletcher-Goldfarb Shanno training algorithm are employed to build the models. The resulting combined mean test set error for the eight local models of 1.31 is significantly better than the mean test set error of 1.85 for the general model. PMID- 11045822 TI - Where are the GaPs? A rational approach to monomer acquisition and selection. AB - Gridding and partitioning (GaP) is a computational method for the classification and selection of monomers for combinatorial libraries. The molecules are described in terms of the pharmacophoric groups they contain and where those pharmacophoric groups can be located in three-dimensional space. The approach involves a detailed conformational analysis of each molecule. This conformational analysis is done within a common coordinate frame, thus enabling the monomers to be compared. The use of a partitioned space is central to this particular application as it facilitates the identification of regions of space which are not well represented by existing compounds. Several ways to extend the use of partitioned pharmacophore spaces are described. Applications of the approach in monomer acquisition and in library design are outlined. PMID- 11045823 TI - E-state modeling of dopamine transporter binding. Validation of the model for a small data set. AB - Data for 25 tropane analogues binding to the dopamine transporter were modeled using E-state molecular structure descriptors. Both E-state and hydrogen E-state descriptors appear in the model in both atom-level and atom-type descriptors. A statistically satisfactory four-variable model is obtained, and structure interpretation is given for each variable, emphasizing substituent influence on nonpolar parts of the molecule as well as the role of hydrogen bonding. A leave group-out approach to model validation is presented in which each observation is removed from the data set three times in random groups of 20% of the whole data set. The average of the resulting predicted values constitutes consensus predictions for these data and supports the claim that the E-state model may be useful for prediction of pIC50 values for new compounds. PMID- 11045825 TI - A cellular automata model of bond interactions among molecules AB - The ten types of alkane bonds have been modeled as isolated fragments using cellular automata dynamics. The rules governing the states and the trajectories of the cells simulating the bonds are derived from the bimolecular interaction accessibilites recently described. The sum of cell encounters at unit iteration (time) becomes a parameter associated with the relationship of a molecule with its neighbors. This value is found to correlate very closely with the boiling points of alkanes, pentanes through octanes. The results reinforce the concept that the molecular connectivity indices are describing the interaction possibilities among alkanes. The study introduces a new way of simulating intermolecular bond encounter dynamics among many molecules. PMID- 11045824 TI - Source of the ice-binding specificity of antifreeze protein type I. AB - Antifreeze proteins (AFPs) are a group of structurally very diverse proteins with the unique capability of binding to the surface of seed ice crystals and inhibiting ice crystal growth. The AFPs bind with high affinity to specific planes of the ice crystal. Previously, this affinity of AFPs has been ascribed to the formation of multiple hydrogen bonds across the protein-ice interface, but more recently van der Waals interactions have been suggested to be the dominant energetic factors for the adsorption. To determine whether van der Waals interactions are also responsible for the binding specificities of AFPs, the protein-ice interaction of the helical AFP Type I from winter flounder (HPLC6) was studied using a Monte Carlo rigid body docking approach. HPLC6 binds in the [1102] direction of the [2021] plane, with the Thr-Ala-Asn surface comprising the protein's binding face. The binding of HPLC6 to this ice plane is highly preferred, but the protein is also found to bind favorably to the [1010] prism plane using a different protein surface comprised of Thr and Ala residues. The results show that van der Waals interactions, despite accounting for most of the intermolecular energy (>80%), are not sufficient to completely explain the AFP binding specificity. PMID- 11045826 TI - The new millennium in health research funding: introducing Canadian Institutes of Health Research. PMID- 11045827 TI - Immunization against bacterial diseases of the intestine. PMID- 11045828 TI - Apoptosis and gastrointestinal disease. PMID- 11045829 TI - Celiac disease in the new world. PMID- 11045830 TI - Vaccines for bacterial enteritis: what is new and why it matters. PMID- 11045831 TI - Improving the outcome for fatty acid oxidation disorders. PMID- 11045832 TI - Evoked potentials for the evaluation of latent hepatic encephalopathy in pediatric liver transplant candidates. AB - BACKGROUND: Visual evoked potentials (VEPs) and brain stem auditory evoked potentials (BAEPs) have been proposed as tools in the diagnosis of subclinical hepatic encephalopathy (HE). However, little information exists to determine their usefulness in pediatric patients. This study was undertaken to evaluate both methods in the detection of subclinical HE in pediatric liver transplant candidates. METHODS: VEPs and BAEPs were recorded in 15 pediatric liver transplant candidates with no clinical signs of HE. The wave latencies found in these examinations were then compared with those in 16 healthy controls of similar age. Laboratory data on liver function and electroencephalographic data from the patients were also recorded to examine their correlation with the evoked potentials results. RESULTS: No differences were found in the BAEP results between patients and controls. However, in the VEPs, the liver transplant candidates had significantly prolonged N1 (N75) latencies when compared with controls; no significant delay was found in the other waves. In contrast, among the children with liver disease, higher BAEP peak latencies correlated positively with electroencephalographic abnormalities, but this correlation was not observed in VEPs. CONCLUSIONS: Evoked potentials might be of use in detecting alterations related to HE in children. However, further studies are necessary to determine their sensitivity and specificity in this situation. PMID- 11045833 TI - Risk factors for severe esophageal and gastric lesions in term neonates: a case control study. Groupe Francophone d'Hepato-Gastroenterologie et Nutrition Pediatrique. AB - BACKGROUND: The purpose of this multicenter case-control study was to search for causes and risk factors related to the severe upper digestive tract lesions often seen in neonates. METHODS: Case patients were full-term neonates with endoscopically confirmed severe bleeding or ulcerative lesions of the esophagus and/or stomach. Matched control subjects were the next infant born in the same maternity unit who met the same criteria and had no clinical abnormality (and, for ethical reasons, no endoscopy). The analysis was based on 137 case-control pairs and considered data showing the mothers' medical and obstetric background, the infants' clinical status and laboratory results, feeding details, and the State and Trait Anxiety Inventory (STAI) questionnaire, which was used to assess the anxiety of the mothers. RESULTS: Cases and controls did not differ in any demographic or social factors. Antacid and anti-ulcer drugs were used significantly more frequently during the last month of pregnancy by case mothers than by control mothers (28% and 10%, respectively; P < 0.001). Mode of delivery was similar. Case infants more frequently experienced cardiac deceleration during labor and delivery (28% and 12.9%; P = 0.003). Breastfeeding at birth was less frequent for case infants (36% and 49%; P = 0.05). The mean trait anxiety scores did not differ between the two groups, but the mean state anxiety score was higher in case mothers. Multivariate logistic regression found that three factors were independently and significantly associated with esophageal and gastric lesions: use of antacid and antiulcer treatments (odds ratio [OR], 3.9; P < 0.001), cardiac deceleration (OR, 2.2; P = 0.03), and breast-feeding (OR, 0.5; P = 0.02). CONCLUSIONS: Antacid drug use by mothers during the last month of pregnancy was associated with esophageal and gastric lesions. Breast-feeding may play a protective role against severe lesions in neonates. PMID- 11045834 TI - Celiac disease in a Chilean population carrying Amerindian traits. AB - BACKGROUND: Although clinical manifestations of celiac disease may change throughout life, clinical, histologic, immunologic, and genetic studies show that there are incomplete forms of this condition, making it difficult to define the disease at a given moment. Because there is no information published in the Latin American-Amerindian population, this study was conducted to assess relations between these parameters in Chileans with celiac disease and their first-degree relatives. METHODS: Sixty-two persons with confirmed celiac disease (mean age, 17.9 +/- 5.1 years; 78.3% females) and 126 relatives (mean age, 27.9 +/- 17.2 years; 65.1% females) were evaluated. Clinical manifestations, antiendomysial antibodies (EMAs), and human leukocyte antigen (HLA) haplotypes were studied in patients. Additionally, jejunal biopsy specimens were assessed (light microscopy) in EMA-positive (EMA+) relatives. RESULTS: Of the patients, 24.1% adhered to a strict gluten-free diet; 26% were oligosymptomatic, and none were malnourished; 45% were EMA+; 13.8% who ingested gluten were EMA-negative (EMA-); one patient consuming a strict gluten-free diet was EMA+. The DQA1*0501 allele was present in the highest frequency (48%, P < 0.0005), whereas combinations of DQ8 were predominant. Of the relatives, 4.8% were EMA+; they had a significantly higher frequency of diarrhea, weight loss, and anorexia (P < 0.03); and all had abnormal histology in biopsy specimens. CONCLUSIONS: After childhood, celiac disease is oligosymptomatic and is often unrecognized by patients. Disease in 13.8% of patients and in 4.8% relatives appeared as incomplete forms of celiac disease. Predominance of DQ8 HLA haplotypes reflects the genetic Spanish-Mapuche heritage of this population. PMID- 11045835 TI - Cutaneous application of vegetable oil as a coadjutant in the nutritional management of preterm infants. AB - BACKGROUND: The cutaneous application of vegetable oil as a therapeutic practice and dietary coadjuvant has been described mainly in adult patients at risk for essential fatty acid deficiency. In the current study, the effects of cutaneous soybean oil application on somatic growth and plasma linoleic and arachidonic acid levels were examined in enterally fed preterm newborns. METHODS: Sixty consecutive preterm infants were chosen from patients admitted to the nursery. Infants were randomly assigned to one of two groups: the oil group, which was treated cutaneously with soybean oil, or the control group, which received no cutaneous treatment. RESULTS: After 30 days, a significant increase in anthropometric parameters was observed in infants who received cutaneous oil, mainly in infants small for gestational age. An increase in linoleic acid level and a decrease in arachidonic acid level were seen in both groups but do not justify the difference found in growth rates in the control and oil groups. CONCLUSIONS: Preterm infants treated cutaneously with soy oil showed better somatic growth than the control group. The factors leading to the present results, especially the response of the infants who were small for gestational age merit further evaluation. PMID- 11045836 TI - Human leukocyte antigen class II alleles in white Brazilian patients with celiac disease. AB - BACKGROUND: Celiac disease (CD) is a permanent gluten intolerance disorder characterized by malabsorption, intestinal mucosa villus atrophy, and crypt hyperplasia. Clinical and histologic features improve in persons consuming a gluten free diet. The pathogenesis of CD involves environmental, genetic, and immunologic factors. METHODS: The frequencies of human leukocyte antigen (HLA) class II alleles were evaluated in white Brazilian patients who had CD and compared with those observed in healthy individuals from the same geographical area (Ribeirao Preto, Sao Paulo) and of similar ethnic background. Twenty-five patients with CD, 11 females and 14 males, and 91 control individuals were studied. The HLA class II alleles were typed using amplified DNA hybridized with sequence-specific primers. Statistical analysis was performed using the two tailed Fisher exact test. The relative risk (RR), etiologic fraction (EF), and preventive fraction (PF) were also estimated. The EF represents the attributable risk for the development of CD at the population level, whereas PF represents the protective risk. RESULTS: The frequency of the HLA-DRB1*03, HLA-DRB1*07, and HLA DQB1*02 alleles was significantly increased in patients. The RR conferred by these alleles was 5.35, 7.15, and 10.6, respectively, and the EF was 48.7%, 44.7%, and 76%, respectively. The frequency of HLA-DQB1*06 alleles was significantly decreased in CD patients, conferring an RR of 0.08 and a PF of 48%. CONCLUSIONS: The results show that HLA-DRB1*03, HLA-DRB1*07, and HLA-DQB1*02 alleles conferred susceptibility to CD in Brazilian patients. In contrast, HLADQB1*06 alleles conferred protection against development of the disease. PMID- 11045837 TI - North American Indian cirrhosis in children: a review of 30 cases. AB - BACKGROUND: North American Indian childhood cirrhosis (NAIC) is a distinct, rapidly evolving form of familial cholestasis found in aboriginal children from northwestern Quebec. This is a retrospective review of the 30 patients treated in Quebec since the discovery of NAIC in 1970. METHODS: The clinical records and histologic samples from 30 patients were reviewed. Extensive metabolic, biochemical, viral, genetic, and radiologic studies were performed in most patients. RESULTS: Genetic analysis suggests autosomal recessive inheritance and a carrier frequency of 10% in this population. Gene mapping studies showed that the NAIC gene is located on chromosome 16q22. Typically, patients have neonatal cholestatic jaundice (70%) or hepatosplenomegaly (20%) with resolution of clinical jaundice by age 1 year but persistent direct hyperbilirubinemia. Portal hypertension was documented in 29 patients (91%). Variceal bleeding (15 patients, 50%) occurred as early as age 10 months. Surgical portosystemic shunting was performed in 13 of these 15 patients (87%); 4 (31%) rebled after 1 to 5 years. Fourteen patients died (47%). In 10 (71%), liver disease was the cause. Four children died of liver failure before liver transplantation became available. In transplanted livers, no recurrence of NAIC was observed after 1 to 10 years. Recognized infectious, metabolic, toxic, autoimmune, and obstructive causes of cirrhosis have been eliminated. The histologic features of NAIC show early bile duct proliferation and rapid development of portal fibrosis and biliary cirrhosis, suggesting a cholangiopathic phenomenon. CONCLUSION: Together with gene mapping studies showing that the NAIC gene is different from those of other familial cholestases, these observations suggest that NAIC is a distinct entity that could be classified as "progressive familial cholangiopathy." PMID- 11045838 TI - Helicobacter pylori infection in children of Texas. AB - BACKGROUND: Acquisition of the Helicobacter pylori infection usually occurs in childhood. The prevalence of infection differs among ethnic groups and in adults is inversely related to the socioeconomic status of the individual's family during childhood. This study investigates the seroprevalence of H. pylori infection in children of different ethnic groups in relation to socioeconomic class and investigates the prevalence of acute H. pylori infection among children who have had recent onset of abdominal pain. METHODS: Serum samples were collected from 797 children, aged 6 months to 18 years, of various socioeconomic and ethnic backgrounds, at a large urban children's hospital. H. pylori status was determined by an anti-H. pylori immunoglobulin (Ig)G enzyme-linked immunosorbent assay (ELISA) validated for pediatric use. To determine the prevalence of acute H. pylori infection, children brought to the emergency center with abdominal symptoms without diarrhea and overt signs of acute abdomen were evaluated with both serology and the 13C-urea breath test. Acute H. pylori was defined as a positive 13C-urea breath test result and negative IgG serology for H. pylori. RESULTS: The overall seroprevalence of H. pylori was 12.2% and increased with age (e.g., 8.3% at 6-11.9 months and 17.9% at 13 years). The prevalence was inversely related to socioeconomic status (6.6%, moderate to high vs. 15%, low socioeconomic status). The difference in seroprevalence among blacks (16.8%), Hispanics (13.3%), and whites (8.3%; P < 0.01) could be accounted for by differences in socioeconomic status. Eighteen percent of children who were evaluated at the emergency center for recent-onset abdominal pain had acute H. pylori infections. CONCLUSIONS: Socioeconomic status, not ethnic group, is the more important risk factor for acquisition of H. pylori infection during childhood. Acute H. pylori infection was a relatively common cause of recent onset, nonsurgical abdominal pain. PMID- 11045839 TI - Clinical presentations and predisposing factors of cholelithiasis and sludge in children. AB - BACKGROUND: In contrast to adults, little is known about the epidemiology and the best therapeutic regimen for cholelithiasis and sludge in children. METHODS: Eighty-two children with cholelithiasis detected by ultrasonography were studied from 0 to 18 years of age with regard to cause, symptomatology, and treatment outcome. Seventy-five children with sludge within the same age group were studied as well. RESULTS: Idiopathic gallstones were found in 19 (23%) patients, and 32 (39%) had gallstones in association with a hemolytic disease. Predominant factors associated with the development of gallstones and clinical presentation differed with age. In patients with sludge, total parenteral nutrition and systemic infection or administration of antibiotics were most frequently found to be possible predisposing factors. Sludge can develop and disappear within a few days. Complications of cholelithiasis were observed in 13 patients. Cholecystectomy was performed in 41 patients and therapeutic endoscopic retrograde cholangiopancreatography (ERCP) with stone extraction in 9 patients; 32 children were not treated. After a follow-up (mean, 4.6 years) in 50 patients, 46% of the children who had cholecystectomy or therapeutic ERCP experienced clinical recurrence of abdominal symptoms. In the patients who did not receive surgical or endoscopic therapy during the follow-up, no complications occurred, and only one patient experienced abdominal symptoms during follow-up. CONCLUSIONS: The difference in associated conditions may indicate that the pathogenesis of cholelithiasis and sludge differ as well. Furthermore, sludge should be viewed as a dynamic condition not predisposing for the development of gallstones, per se. Cholecystectomy should not be performed routinely but only after careful selection in patients at risk for complications. PMID- 11045840 TI - Energy supplements rich in linoleic acid improve body weight and essential fatty acid status of cystic fibrosis patients. AB - BACKGROUND: Patients with cystic fibrosis who have steatorrhea frequently are underweight and have essential fatty acid (EFA) depletion, which is associated with a poor clinical course. It has been stated that poor EFA status is difficult to correct in patients with cystic fibrosis, and an impaired EFA metabolism with reduced synthesis of long-chain polyunsaturated fatty acids has been proposed. In this study, the effects of an oral energy supplement rich in linoleic acid were investigated in patients with cystic fibrosis who had a body weight below 95% of normal for height. METHODS: Thirty-six patients (16 girls) more than 4 years of age were randomized either to a control group (n = 20, age 13.3 +/- 3.8 years, mean +/- SD) receiving intensive dietary counseling only, or an intervention group (n = 16, age, 10.4 +/- 4.3 years) treated for 3 months with dietary counseling plus 628 +/- 254 mL (= kcal) per day of an energy supplement rich in fat (31% of energy) and linoleic acid (16% of energy). RESULTS: In contrast to the control group, the patients with supplemented diets achieved significant increases of energy intake (2189 +/- 731 kcal/day vs. 2733 +/- 762 kcal/day), weight for height (82.8% +/- 8.6% vs. 84.8% +/- 9.6% of normal), and body fat (5.1 +/- 1.7 kg vs. 5.8 +/- 2.2 kg) as well as the initially low values of plasma phospholipid linoleic acid (11.8% +/- 1.1% vs. 17.6% +/- 1.6% of total phospholipid fatty acids) and its main metabolite arachidonic acid (4.4% +/- 0.4% vs. 5.9% +/- 0.3%). CONCLUSIONS: Patients with cystic fibrosis with low body weight and poor EFA status benefit from EFA-rich energy supplements and can synthesize arachidonic acid from the precursor linoleic acid. PMID- 11045841 TI - Helicobacter pylori infection in recurrent abdominal pain. AB - BACKGROUND: Helicobacter pylori is associated with chronic gastritis and peptic ulcer in adults and in children. The purpose of the present study was to analyze the association of recurrent abdominal pain and H. pylori infection in children and to evaluate the efficacy of antimicrobial treatment in patients with evidence of infection. METHODS: The clinical and histopathologic findings in children who underwent diagnostic upper endoscopy for recurrent abdominal pain were analyzed retrospectively. Patients with evidence of infection with H. pylori were treated with a combination of omeprazole, amoxicillin, and clarithromycin. Efficacy of treatment was assessed using the 13C-urea-breath test. RESULTS: H. pylori was found in histopathologic sections of 29 (40%) of 73 patients undergoing diagnostic endoscopy for recurrent abdominal pain. Five children (17%) were of Swiss ethnic origin, and 24 (83%) were non-Swiss. All the infected patients had chronic gastritis and 4 (14%) had ulcerations in the duodenum. Treatment with omeprazole, amoxicillin, and clarithromycin resulted in eradication of the infection in all and in resolution of the clinical symptoms in 15 (80%) of 19 patients who had a follow-up examination. CONCLUSIONS: The presented data suggest that gastritis induced by H. pylori may be associated with recurrent abdominal pain and that in Switzerland infections with H. pylori primarily involve persons who are non-Swiss. A combined therapy results in eradication of the bacterium and in improvement of the clinical symptoms in a significant majority of the patients. PMID- 11045842 TI - Measurement of total body water in children using bioelectrical impedance: a comparison of several prediction equations. AB - BACKGROUND: Body composition evaluation by bioelectrical impedance analysis in children makes use of different group-specific population-derived equations. The present study was conducted to attempt to validate the use of population independent physical model-derived equations in children. METHODS: The validity of bioelectrical impedance analysis for the measurement of total body water in children was evaluated by comparing results of two physical model-derived and two population-derived equations with those of deuterium dilution as reference method in a group of 38 heterogeneous children. RESULTS: Means +/- standard deviation (in liters) for total body water measured with deuterium dilution and the physical model 1-derived equation were 18.4 +/- 4.7 L and 18.1 +/- 4.4 L, respectively. This difference is not significant, whereas significant differences were found for all other tested equations. Significant smaller absolute differences between the model 1 equation and deuterium reference results were found when compared with the results of the other three tested equations. CONCLUSION: When compared with results of the reference deuterium method the physical model 1-derived equation was the only one that provided reliable total body water results by bioelectrical impedance analysis in children. PMID- 11045844 TI - Nonobstructive antral web: an unusual cause of excessive crying in an infant. PMID- 11045843 TI - Lactose-[13C]ureide breath test: a new, noninvasive technique to determine orocecal transit time in children. AB - BACKGROUND: The lactose-[13C]ureide breath test (LUBT) is a novel, noninvasive test to determine orocecal transit time. Lactose ureide resists the action of brush border enzymes and is metabolized by colonic bacteria. The purpose of the present study was to adapt this breath test for children of various age groups and to determine whether it can be applied in infants, newborns, and preterms to study the development of small intestinal motility. METHODS: In a group of 20 children (3-17 years) in vitro stool analysis and in vivo LUBT results were compared. From each subject a blank stool sample and a sample produced after induction with unlabeled lactose ureide were incubated with 10 mg lactose [13C]ureide in small, closed bottles. Ten-milliliter CO2 samples were aspirated from the bottles using a needle and a syringe every 30 minutes for 24 hours. All children performed the breath test after induction of 500 mg unlabeled lactose ureide three times the prior day. A liquid test meal (chocolate milk) with 250 mg lactose-[13C]ureide was ingested. Breath samples were collected every 15 minutes for 10 hours. In a second group of 32 children (age range, 0-3 years) consisting of 6 children between 1 and 3 years of age, 6 infants between 6 and 12 months, 13 infants between 0 and 6 months, and 7 preterm infants, only the in vitro stool analysis was performed. Stools were collected for stool incubation, as described. RESULTS: The mean orocecal transit time in the group of 20 children aged 3 to 17 years was 255 minutes (range, 165-390 minutes). Stool incubations demonstrated a clear 13CO2 peak in all infants aged more than 8 months, indicating that their colonic bacterial enzymic activity hydrolyses lactose ureide. However, in all infants aged less than 6 months and in preterm infants, the 13CO2 signal was absent, indicating that those subjects were unable to hydrolyze lactose ureide. CONCLUSION: Infants aged less than 6 months do not host the appropriate bacterial enzymic activity for splitting lactose ureide. The authors conclude that the LUBT can be applied in infants aged more than 8 months, after weaning to solid foods, to determine orocecal transit time. PMID- 11045845 TI - Use of a gastrostomy tube to perpetrate Munchausen syndrome by proxy. PMID- 11045846 TI - Somatostatin in the treatment of pancreatic pseudocyst complicating acute pancreatitis in a child with liver transplantation. PMID- 11045847 TI - Complications in early diagnosis and treatment of two infants with long-chain fatty acid beta-oxidation defects. PMID- 11045848 TI - Is lactobacillus GG helpful in children with Crohn's disease? Results of a preliminary, open-label study. AB - BACKGROUND: Lactobacillus GG is a safe probiotic bacterium known to transiently colonize the human intestine. It has been found to be useful in treatment of several gastrointestinal conditions characterized by increased gut permeability. In the current study, the efficacy of Lactobacillus GG was investigated in children with Crohn's disease. METHODS: In this open-label pilot evaluation viewed as a necessary preliminary step for a possible subsequent randomized placebo-controlled trial, four children with mildly to moderately active Crohn's disease were given Lactobacillus GG (10(10) colony-forming units [CFU]) in enterocoated tablets twice a day for 6 months. Changes in intestinal permeability were measured by a double sugar permeability test. Clinical activity was determined by measuring the pediatric Crohn's disease activity index. RESULTS: There was a significant improvement in clinical activity 1 week after starting Lactobacillus GG, which was sustained throughout the study period. Median pediatric Crohn's disease activity index scores at 4 weeks were 73% lower than baseline. Intestinal permeability improved in an almost parallel fashion. CONCLUSIONS: Findings in this pilot study show that Lactobacillus GG may improve gut barrier function and clinical status in children with mildly to moderately active, stable Crohn's disease. Randomized, double-blind, placebo-controlled trials are warranted for a final assessment of the efficacy of Lactobacillus GG in Crohn's disease. PMID- 11045849 TI - Clinical quiz. Hepatitis A. PMID- 11045850 TI - Posttransplantation lymphoproliferative disorder: endoscopic findings. AB - BACKGROUND: Posttransplantation lymphoproliferative disorder (PTLD) may manifest a variety of nonspecific symptoms and must be suspected in the patient who undergoes solid organ transplantation. Common sites of occurrence include the gastrointestinal tract, the central nervous system, and lymphoid tissue of the oral pharynx, mediastinum, and mesentery. The large incidence of gastrointestinal involvement provides an opportunity for endoscopic diagnosis. This is the description of a characteristic endoscopic finding in patients who have undergone liver transplantation who are under evaluation for suspected PTLD. METHODS: During a 2-year period, 27 liver transplantations were performed in 24 pediatric patients. Fourteen patients underwent endoscopic evaluation. Indications for endoscopy included abdominal pain, vomiting, hematemesis, irritability, growth failure, anemia, occult blood loss, and suspected PTLD. Biopsy specimens were obtained from any endoscopically detected abnormality and from the duodenum, gastric antrum, esophagus, terminal ileum, cecum, and rectum. Specimens with suspected PTLD were evaluated with Epstein-Barr virus latent membrane stain. RESULTS: Six patients were found to have a characteristic lesion, which was raised, rubbery, and erythematous, with a central ulceration. Lesions were singular or multiple and ranged from 5 to 15 mm in diameter. Microscopic evaluation revealed a monotonous proliferation of lymphocytes. All specimens were positive for Epstein-Barr virus latent membrane protein stain. Stains for cytomegalovirus were negative. Biopsy specimens from the eight patients without identified characteristic lesions were negative for PTLD. CONCLUSIONS: Panendoscopy is a useful tool for the diagnosis and treatment of gastrointestinal PTLD. Endoscopy is easily accomplished, may provide an instantaneous result if the characteristic lesion is identified, and provides tissue for disease classification. Patients with unexplained gastrointestinal signs or symptoms should undergo panendoscopy for suspected PTLD. PMID- 11045851 TI - Feeding preterm infants after hospital discharge: effect of diet on body composition. PMID- 11045852 TI - The Research Society on Alcoholism (RSA): 1976-2000. PMID- 11045853 TI - Polymorphism in the interleukin-1 receptor antagonist gene is associated with alcoholism in Spanish men. AB - BACKGROUND: A polymorphism located in intron 2 of the interleukin-1 receptor antagonist (IL1RN) gene recently has been associated with the development of hepatic fibrosis in Japanese alcoholics. In the present study, we analyzed whether there is an association between this polymorphism, alcoholism, and alcoholic liver disease in a Spanish male population of alcoholics. METHODS: The IL1RN genotype was assessed by polymerase chain reaction by using oligonucleotides that flank a variable nucleotide tandem repeat polymorphism located in intron 2 of this gene in 90 male alcoholic patients from Spain: 30 alcohol-dependent men, 30 alcohol abusers, and 30 alcoholics with liver cirrhosis. We also studied 40 healthy subjects. RESULTS: The distribution of the IL1RN allelic frequencies in Spanish healthy subjects is similar to that previously reported in White subjects. However, the A1 allele is overrepresented in Spanish alcoholics when compared with healthy subjects. No significant differences in allelic frequencies were observed between alcoholics with liver cirrhosis and alcoholics without liver disease or between alcohol-dependent subjects and alcohol abusers. CONCLUSION: The presence of the A1 allele of the IL1RN gene is associated with a higher risk of alcoholism in Spanish men. PMID- 11045854 TI - Heritability of the blood pressure response to acute ethanol exposure in five inbred strains of mice. AB - BACKGROUND: Chronic alcohol consumption is a major risk factor for hypertension. There is evidence in humans that the susceptibility to alcohol-related hypertension may vary based on genotype. As a first step in investigating the genetic basis for alcohol-related hypertension, the current study was designed to assess the heritability of the blood pressure response to acute ethanol exposure by using AKR/J (AK), C57BL/6J (B6), DBA/2J (D2), Balb/cJ (Balb), and A/J (A) mice. METHODS: Mean arterial pressure (MAP) was recorded continuously for 24 hr in freely moving mice from an indwelling femoral catheter before we tested the effects of saline or ethanol (2 g/kg ip) on blood pressure. RESULTS: Relative to saline, ethanol caused a pressor response that peaked within 10 min, followed by a decline in MAP. Strain A mice had a significantly greater pressor response to ethanol than other strains and did not show a decline in MAP below baseline. All other strains showed a progressive fall in blood pressure below baseline across the 60 min measurement interval. Heritability was estimated to be 0.62 for the pressor response and 0.64 for the maximal depressor response. Repeated doses of ethanol at 1 hr intervals in A and B6 mice (0, 2, 1.5, 1.5, 1.5 g/kg ip) resulted in a dose-dependent increase in MAP in A mice for the first three doses and a dose-dependent decrease in MAP in B6 mice that was independent of blood ethanol concentrations. CONCLUSION: The results indicate that there is a significant genetic component to the acute blood pressure response to ethanol. PMID- 11045855 TI - Differences in the fatty acid composition of fatty acid ethyl esters in organs and their secretions. AB - BACKGROUND: Fatty acid ethyl esters (FAEE) are nonoxidative ethanol metabolites that have been shown to be long term markers of ethanol intake and have been implicated as mediators of ethanol-induced cell injury. Previous studies have indicated that the fatty acid composition of the FAEE found in the plasma of human subjects after ethanol ingestion is predominantly ethyl palmitate and ethyl oleate. This raised the possibility that there is some selectivity toward the fatty acid used for FAEE to be exported from the liver into the blood. METHODS: To address the hypothesis that the fatty acid composition of FAEE secreted from organs, such as the liver and pancreas, differs from the fatty acid composition of FAEE in the organs, this study was performed using rats that received ethanol by intra-arterial infusion. RESULTS: It was found that the fatty acids in FAEE differed significantly in plasma versus liver, bile versus liver, and pancreatic secretions versus pancreas. CONCLUSIONS: These results indicate that organs selectively export certain FAEE species. PMID- 11045856 TI - Neurophysiological findings and drinking levels in high-alcohol-drinking (HAD) and low-alcohol-drinking (LAD) rats. AB - BACKGROUND: Specific neurophysiological profiles, such as reduced P300 amplitude or altered spectral power in the EEG, have been associated with a risk for alcoholism in several clinical populations. In certain rodent models, high versus low alcohol consumption is associated with similar neurophysiological differences. For example, alcohol-preferring (P) rats have increased spectral power and decreased P300 amplitudes compared with alcohol-nonpreferring (NP) rats. In the present study, the neurophysiological profiles of high-alcohol drinking (HAD) and low-alcohol-drinking (LAD) rats were assessed (1) to determine if their electrophysiological profiles are similar to P and NP rats and (2) to examine the relationship of these neurophysiological indices to ethanol drinking. METHODS: Ethanol-naive HAD and LAD rats were implanted with cortical and amygdalar recording electrodes. Baseline EEG and event-related potentials (ERPs) then were assessed. Subsequently, all rats were trained to self-administer ethanol by using a sucrose-substitution procedure. RESULTS: Baseline EEG and ERP (i.e., pre-ethanol exposure) were assessed based on line (HAD versus LAD) and actual ethanol consumption (high drinkers versus low drinkers). At baseline, ethanol-naive HAD rats displayed significantly greater power in the cortical EEG and decreased amygdala N1 ERP amplitude compared with ethanol-naive LAD rats. Similar EEG and ERP profiles have been observed when P and NP rats are compared. No differences in P300 between lines were observed, but high-drinking rats, independent of line, had significantly decreased P300 amplitude in the amygdala compared with low-drinking rats. CONCLUSIONS: These data suggest there are some similarities in EEG and ERP profiles of P and HAD rats compared with NP and LAD rats. Furthermore, the data suggest that decreased P300 amplitude in the amygdala is associated with increased limited access ethanol drinking. PMID- 11045857 TI - Effects of MDL 72222, a serotonin3 antagonist, on operant responding for ethanol by alcohol-preferring P rats. AB - BACKGROUND: Previous studies indicated that, under continuous access conditions, the 5-HT3 antagonist MDL 72222 (MDL) effectively reduced ethanol drinking of alcohol-preferring P rats. However, MDL was without effect when similar doses were tested under scheduled access conditions, unless the ethanol access period was randomly presented. This study examined the effects of MDL on operant responding for ethanol and water by adult male alcohol-preferring P rats. METHODS: During the dark cycle, subjects in the first experiment were trained to respond concurrently for 15% ethanol and water on a fixed-ratio 5 (FR-5) and FR-1 schedule of reinforcement, respectively. Approximately 30 min before the 4-hr operant session, rats were injected subcutaneously (sc) with saline or MDL (1, 3, or 5 mg/kg). A second experiment tested the effects of 1 mg/kg MDL on operant responding for 15% ethanol in 1-hr sessions when operant access was given at a fixed time each day (fixed scheduled access, FSA group) or at variable time periods throughout the dark cycle (variable scheduled access, VSA group). RESULTS: In the first experiment, only the 5 mg/kg dose of MDL decreased responding for ethanol (approximately 20%) during the first 30 min of the 4-hr session. This dose also reduced total 4-hr responding for ethanol and water. In the second experiment, the 1 mg/kg dose of MDL had no effect on operant responding by the FSA group, but significantly reduced ethanol responding by the VSA group. CONCLUSIONS: These results suggest that 5-HT3 receptors may be involved in mediating the reinforcing effects of ethanol, and that temporal environmental cues associated with the presentation of ethanol may be one factor involved in reducing the effectiveness of 5-HT3 antagonists to attenuate ethanol intake. PMID- 11045858 TI - Effect of hormone balance on carbohydrate-deficient transferrin and gamma glutamyltransferase in female social drinkers. AB - BACKGROUND: Carbohydrate-deficient transferrin (CDT) and gamma glutamyltransferase (GGT) are the most commonly used markers for alcohol abuse, but their sensitivity and specificity are lower and have different reference values among females compared with males. In the present study, we evaluated the effect of women's hormone balance on these two alcohol markers, as well as on their mathematical combination, named gamma-CDT. METHODS: An age-stratified random sample of 3962 women, between 25-74 years old, was drawn from the normal population. Pregnancy, the use of oral contraceptives, intrauterine device for contraception, hormone replacement therapy, and hormone treatment for infertility were considered. A comparison between fertile, peri- and postmenopausal women was also done. RESULTS: Existing pregnancy increased CDT levels but decreased GGT values. Lower CDT and higher GGT levels were observed among those women using oral contraceptives and in postmenopausal women compared with women at the fertile stage. gamma-CDT was not influenced by hormone balance. CONCLUSIONS: The different hormonal status had an opposite effect on CDT and GGT. Women who were close to late menopause had levels of both markers closer to the values of men. It must be pointed out that the findings presented here are based on measurements of absolute CDT values and that no measurements of total transferrin were done. gamma-CDT, not influenced by hormone balance, indicates promising clinical utility among women. PMID- 11045859 TI - The neuropsychological consequences of abstinence among older alcoholics: a cross sectional study. AB - BACKGROUND: The older alcoholic has been distinguished from the younger alcoholic with regard to both the acute effects of alcohol and also the recovery of functioning with abstinence. Few studies, however, have included samples of exclusively older subjects. In this investigation we examined the recovery of functioning in an older cohort of recovering alcoholics (age range 55-83) to determine which neuropsychological functions improve and which remain impaired with abstinence. METHODS: We used a cross-sectional design, comparing three demographically matched groups on a battery of neuropsychological tests: (a) older alcoholics who had been abstinent for greater than 6 months, (b) older alcoholics who had been abstinent for less than 6 months, and (c) a control group of older subjects without alcohol abuse histories. RESULTS: In almost all tasks, the alcoholics who were abstinent for less than 6 months performed worse than the control group. In contrast, the alcoholics who had been abstinent for more than 6 months differed from the control group on learning and recall of a word list, immediate and delayed recall of a complex figure, initial letter fluency, and clock drawing. CONCLUSIONS: Memory and executive skills appear to be resistant to recovery or at least slower to recover with abstinence in the older alcoholic. The impairment with visuospatial skills reported in prior investigations of alcoholics may be related to compromised executive functions, which interfere with the encoding of more complex visuospatial information and thus affect recall of such information. Studies that involve larger samples of older alcoholics are needed to understand their ability to recover cognitive functioning with abstinence. PMID- 11045860 TI - Treatment of problem alcohol use in women of childbearing age: results of a brief intervention trial. AB - BACKGROUND: Studies suggest that 14% of women age 18 to 40 drink alcohol above recommended limits. Of special concern is the increasing use of alcohol by women during pregnancy. This article reports 48 month follow-up data from a subanalysis of a trial for early alcohol treatment (Project TrEAT) focused on women of childbearing age. METHODS: Project TrEAT was conducted in the offices of 64 primary care, community-based physicians from 10 Wisconsin counties. Of 5979 female patients ages 18 to 40 who were screened for problem drinking, 205 were randomized into an experimental group (n = 103) or control group (n = 102). The intervention consisted of two 15 min, physician-delivered counseling visits that included advice, education, and contracting by using a scripted workbook. A total of 174 subjects (85%) completed the 48 month follow-up procedures. RESULTS: No significant differences were found between the experimental and control groups at baseline for alcohol use, age, socioeconomic status, smoking, depression or anxiety, conduct disorder, lifetime drug use, or health care utilization. The trial found a significant treatment effect in reducing both 7 day alcohol use (p = 0.0039) and binge drinking episodes (p = 0.0021) over the 48 month follow-up period. Women in the experimental group who became pregnant during the follow-up period had the most dramatic decreases in alcohol use. A logistic regression model based on a 20% or greater reduction in drinking found an odds ratio of 1.93 (confidence interval 1.07-3.46) in the sample exposed to physician intervention. Age, smoking, depression, conduct disorder, antisocial personality disorder, and illicit drug use did not reduce drinking significantly. No significant differences were found in health care utilization and health status between groups. CONCLUSIONS: This trial provides the first direct evidence that brief intervention is associated with sustained reductions in alcohol consumption by women of childbearing age. The results have enormous implications for the U.S. health care system. PMID- 11045861 TI - Predictors of substance abuse treatment retention among women and men in an HMO. AB - BACKGROUND: Although prior research has examined predictors of treatment retention in public alcohol and drug treatment programs, little is known about factors that influence treatment retention in an insured outpatient population. Because there is growing evidence that the factors which influence treatment retention may differ by gender, we identify sex-specific predictors. METHODS: We recruited all eligible intakes to a health maintenance organization's outpatient alcohol and drug treatment program during a 2-year period and obtained a sample of 317 women and 599 men. The programs, day hospital and traditional outpatient modalities, were abstinence based. We separated our sample by sex and used least squares and logistic regression to identify independent predictors of length of stay and program completion, respectively. RESULTS: One general pattern of predictors of increased retention was shared by women and men in this alcohol and drug treatment program--fewer and less severe drug problems. However, most predictors were sex-specific. Among women, retention was predicted by having higher incomes, belonging to ethnic categories other than African American, being unemployed, being married, and having lower levels of psychiatric severity. Among men, predictors of higher retention included being older, receiving employer suggestions to enter treatment, and having abstinence goals. CONCLUSIONS: These findings highlight the importance of examining aspects of the course of treatment separately by sex. They also suggest treatment factors that may enhance retention among insured populations, including employer referrals, psychiatric services, and drug-related services. PMID- 11045862 TI - Efficacy of dexfenfluramine in the treatment of alcohol dependence. AB - BACKGROUND: A substantial body of evidence supports a role for serotonin in modulating alcohol intake, which suggests that this neurotransmitter represents a promising target for pharmacotherapy development for alcohol use disorders. Dexfenfluramine. a serotonin releaser and reuptake inhibitor, decreases alcohol self-administration by rats. Its greater potency and several mechanisms of action suggest it should be more effective in treating alcohol dependence than drugs that only inhibit serotonin reuptake. METHODS: We conducted an 11 week, randomized, double-blind trial that compared oral placebo and dexfenfluramine 7.5, 15, 22.5, and 30 mg bid in 136 alcohol-dependent patients. A brief behavioral intervention was offered concurrently. RESULTS: The majority of subjects were male (72%), and the age of the group was 44 +/- 1 years (mean +/- SD). Both placebo- and drug-treated groups significantly reduced alcohol consumption compared with baseline (a 55% decrease in mean drinks per day; p < 0.01), but there were no significant differences between drug and placebo groups or dose effects for most outcome measures. CONCLUSIONS: Our results with dexfenfluramine are further evidence that serotonergic medications on their own do not significantly reduce alcohol consumption in alcohol-dependent individuals. Combination pharmacotherapy with agents that act on different receptors or neurotransmitter systems (e.g., naltrexone plus dexfenfluramine) may be one way to enhance serotonergic effects on drinking behavior and should be considered in future medication development clinical trials. PMID- 11045863 TI - Predictors of compliance with naltrexone among alcoholics. AB - OBJECTIVE: Naltrexone has been found to be an effective adjunct to treatment to reduce the rate of drinking among alcoholics. However, adherence to the medication has been of considerable concern; the high rates of noncompliance with the medication limits the benefits that could potentially be realized from this pharmacotherapy. Knowledge of predictors of noncompliance could result in interventions targeted at these variables. METHOD: Participants were 128 alcohol dependent patients who participated in a clinical placebo-controlled trial of naltrexone. Upon discharge from a 1- to 2-week partial hospital program, patients were randomly placed into 12 weeks of naltrexone (50 mg/day) or placebo (n = 64 per condition). Patients met with a physician and a research assistant weekly for 4 weeks then biweekly for 8 weeks. RESULTS: Compliance (number of days taking medication) was not predicted by demographic or pretreatment alcohol use variables. Number and severity of side effects in the first week, particularly nausea and fatigue, predicted early termination. Compliance was not predicted by commitment to abstinence or self-efficacy about abstinence, but was greater among patients who believed more strongly that the medication would help them stay sober. Compliance was not predicted by general level of urge to drink during the first week on medication but compliance was greater among those with a higher urge to drink in response to alcohol stimuli in the laboratory. CONCLUSIONS: Implications for approaches to increase compliance include reducing side effects and increasing patients' beliefs in the efficacy of naltrexone. PMID- 11045864 TI - Ethanol exposure enhances apoptosis within the testes. AB - BACKGROUND: Chronic ethanol abuse causes testicular atrophy and male infertility in alcoholic men. It is well known that ethanol exposure disrupts the hypothalamic-pituitary-gonadal axis, adversely affects the secretory function of Sertoli cells, and produces oxidative stress within the testes. It is still not clear what cellular mechanisms are responsible for the morphologic alteration of the testes that results in a reduction of testicular mass as a consequence of ethanol exposure. The hypothesis tested was that ethanol enhances apoptosis of testicular germ cells. METHODS: In the experiments of chronic ethanol exposure, male Sprague Dawley rats (Harlan Sprague Dawley, Inc., Indianapolis, IN) were fed Liber-Decarlie liquid diet for 9 weeks. In the experiments of acute ethanol exposure, a small volume of 20% ethanol solution was administered by intratesticular injection. Both 3'-end labeling of isolated testicular deoxyribonucleic acid (DNA) and labeling of apoptotic cells in situ by the terminal deoxynucleotidyl transferase-mediated deoxyuridine 5'-triphosphate nick end-labeling method were used to determine apoptosis rates within the testes. The expression of proteins involved in apoptosis was assessed by reverse transcription-polymerase chain reaction and by Western blotting. RESULTS: The testes of rats that were fed an ethanol-containing liquid diet had more testicular DNA fragmentation than did those of animals that were fed an isocaloric control diet. Ethanol increased the number of apoptotic spermatogonia as well as spermatocytes. Direct intratesticular injections of ethanol solution enhanced testicular DNA fragmentation, suggesting an increase in apoptosis. Moreover, Fas ligand levels were increased within the testes of rats that were chronically fed ethanol. In vitro, ethanol treatment of cultured Sertoli cells enhanced the production of Fas ligand. In addition, testicular levels of p53 messenger ribonucleic acid were increased in rats that were chronically fed ethanol. CONCLUSIONS: All of these observations suggest that ethanol enhances testicular germ cell apoptosis. PMID- 11045865 TI - Apoptosis and dysregulated ceramide metabolism in a murine model of alcohol enhanced lipopolysaccharide hepatotoxicity. AB - BACKGROUND: The role of apoptosis in EtOH-induced liver injury has not been investigated much. Therefore, the question whether apoptosis is a contributory factor to alcoholic liver disease remains to be answered. The purpose of this study was to characterize the liver apoptotic response in a murine model of alcohol-enhanced lipopolysaccharide (LPS) hepatotoxicity. METHODS: Mice were fed an alcohol-containing liquid diet for 49 days followed by an acute LPS challenge. The liver state was judged on the basis of histological appearance, plasma liver enzyme activity (alanine:2-oxoglutarate and aspartate:2-oxoglutarate aminotransferases, as markers of hepatocytolysis), and plasma hyaluronan levels (as a marker of the sinusoidal endothelial cell scavenging function). The liver apoptotic response was assessed by DNA fragmentation (TUNEL procedure), and caspases-3 and -8 activity. To determine if ceramide played a role in the liver apoptotic response, the activity of acidic sphingomyelinase and tissue content of ceramide were also quantified. RESULTS: Alcohol exposure induced fat accumulation and sensitized the liver to LPS injurious effects. Plasma liver enzyme activity was elevated by alcohol and this effect was potentiated by LPS. Liver apoptosis was augmented by both alcohol and LPS treatment as reflected by high frequency of positive TUNEL staining nuclei and by an increased activity of caspase-3 and -8. Acidic sphingomyelinase activity was also increased and it was associated with an elevated tissue content of ceramide. In addition, LPS also increased plasma TNF alpha levels. These changes were accompanied by elevated plasma hyaluronan, reflecting an impaired sinusoidal endothelial cell scavenging function. CONCLUSIONS: These results provide a more complete description of the liver apoptotic response to both alcohol and LPS and may constitute the basis for further mechanistic studies on a possible role apoptosis may play in alcoholic liver injury. PMID- 11045866 TI - Postnatal handling does not attenuate hypothalamic-pituitary-adrenal hyperresponsiveness after prenatal ethanol exposure. AB - BACKGROUND: Prenatal ethanol exposure results in hypothalamic-pituitary-adrenal (HPA) hyperresponsiveness to stress in the adult animal. In contrast, an early environmental manipulation, termed "postnatal handling," has been shown to result in decreased and/or less prolonged HPA activity in response to moderate stressors throughout the lifespan of the animal. The effects of both prenatal ethanol exposure and postnatal handling on HPA activity may be mediated by altered feedback regulation of the HPA axis. The present study tested the hypothesis that postnatal handling could attenuate the impact of prenatal ethanol exposure on hormonal responses to stressors. METHODS: Male and female Sprague Dawley rats from prenatal ethanol (E), pair-fed (PF), and ad libitum-fed control (C) groups were either handled (H) or nonhandled (NH) during the preweaning period and were tested at 4 to 5 months of age. Animals were subjected to a 60 min restraint stress, 3 hr after intraperitoneal injection with either saline (SAL) or a synthetic glucocorticoid, dexamethasone-21-phosphate (DEX), in order to examine HPA responsiveness after DEX blockade of endogenous HPA activity. Blood samples were collected via jugular cannulae immediately before restraint (0 min), during restraint (10, 30, and 60 min), and 30 min after the termination of restraint (90 min). RESULTS: For both males and females, DEX administration significantly reduced plasma adrenocorticotropic hormone (ACTH) and corticosterone (CORT) concentrations compared with SAL administration. H animals showed greater suppression of HPA activity (i.e., lower ACTH and/or CORT levels) than NH animals regardless of prenatal group. In addition, E females from both the H and NH treatments showed elevated ACTH and CORT after both SAL and DEX administration, whereas H and NH E males showed elevations in ACTH and CORT only after SAL, compared with their PF and C counterparts. CONCLUSIONS: These data extend results from previous studies that demonstrated HPA hyperresponsiveness in E animals. The finding that E females but not males exhibit elevated ACTH and CORT after DEX administration suggests that prenatal ethanol exposure results in sex-specific alterations of HPA feedback. Consistent with previous data, handling in itself reduced the HPA response to restraint stress. However, handling did not attenuate either HPA hyperresponsiveness or feedback deficits in E animals. PMID- 11045867 TI - Reduced hypothalamic POMC and anterior pituitary CRF1 receptor mRNA levels after acute, but not chronic, daily "binge" intragastric alcohol administration. AB - BACKGROUND: Endogenous corticotropin-releasing factor (CRF), its pituitary CRF1 receptor, and proopiomelanocortin (POMC) may be involved in the hypothalamic pituitary-adrenal (HPA) responses to alcohol. METHODS: Alcohol (1.5 g/kg) or water was administered intragastrically to male Fischer rats after the "binge" pattern regimen, that is, three times daily at 1 hr intervals at the beginning of the light cycle. The levels of CRF, CRF1 receptor, and POMC mRNAs in the hypothalamic-pituitary axis were measured after acute (1 day) or chronic (14 days) binge pattern alcohol administration. Plasma levels of ACTH and corticosterone were measured to examine time-dependent alterations of HPA responses. RESULTS: Plasma ACTH and corticosterone levels were elevated dramatically after 1 day of acute binge pattern alcohol administration. After 14 days of chronic alcohol, however, no elevation in plasma ACTH levels and an attenuated elevation in plasma corticosterone levels were found. CRF mRNA levels in the hypothalamus were not altered after either acute or chronic alcohol administration. CRF1 receptor mRNA levels in the anterior pituitary were decreased significantly after acute administration, with no change after chronic alcohol administration. POMC mRNA levels in the anterior pituitary were not altered by either acute or chronic alcohol administration. In the hypothalamus, POMC mRNA levels were decreased significantly after acute but not chronic binge alcohol administration. CONCLUSIONS: These results suggest that (1) rats exposed to chronic binge alcohol develop tolerance in HPA activity, as shown by no elevation of ACTH and an attenuated corticosterone response to chronic alcohol after initial dramatic elevations by acute alcohol administration; (2) a concurrent acute decrease in CRF1 receptor mRNA levels in the anterior pituitary is associated with increased HPA activity, and (3) alterations of POMC gene expression in the hypothalamic region may have implications for a molecular understanding of the neuroendocrine response to alcohol. PMID- 11045868 TI - Chronic, but not acute, nicotine exposure attenuates ethanol withdrawal-induced hippocampal damage in vitro. AB - BACKGROUND: Long-term ethanol use and long-term tobacco use frequently occur together, which suggests concurrent dependence on ethanol and nicotine. Consequences of this form of polydrug dependence are not well understood, however. Previous evidence suggests detrimental effects of long-term ethanol and beneficial effects of nicotine exposure on neuronal viability. Thus, the present study was designed to use an organotypic hippocampal slice culture model to examine the ability of chronic and acute nicotine exposure to reduce neurotoxicity associated with withdrawal from long-term ethanol exposure. METHODS AND RESULTS: Twenty-four hours of withdrawal after continuous 10 day ethanol exposure (50 or 100 mM in culture medium) resulted in cytotoxicity in hippocampal slice explants obtained from neonatal rat, most notably in pyramidal cell layers of the CA1 region. Exposure of slices to the N-methyl-D-aspartate receptor blocker MK-801 during ethanol withdrawal significantly reduced this toxicity. Exposure of slices to nicotine (0.1-10.0 microM) during the 24 hr withdrawal period did not reduce hippocampal damage. However, treatment of slices with nicotine (0.1-10.0 microM) during 10 days of ethanol exposure was associated with significant reductions in subsequent withdrawal-induced cytotoxicity, an effect reduced by mecamylamine coexposure with nicotine and ethanol. CONCLUSIONS: These findings indicate the development of marked hippocampal neurotoxicity during withdrawal from long-term ethanol exposure that is mediated, in part, by overactivation of N-methyl-D-aspartate receptors. Furthermore, these data suggest that one consequence of concurrent dependence on ethanol and nicotine may be reduced neurological damage associated with ethanol withdrawal. PMID- 11045869 TI - 6-beta-naltrexol reduces alcohol consumption in rats. AB - BACKGROUND: In humans, 6-beta-naltrexol is the major metabolite of naltrexone, and its effectiveness at suppressing alcohol consumption in any species has not been previously investigated. Naltrexone is an opiate antagonist that reduces excessive drinking in many species, including humans with alcohol dependence. Whether 6-beta-naltrexol is an active metabolite that contributes to the efficacy of naltrexone remains unknown. METHODS: Placebo and four doses of 6-beta naltrexol were given by intraperitoneal injection to outbred Wistar rats and alcohol consumption was measured using a limited access model. RESULTS: 6-beta Naltrexol reduced alcohol consumption in a dose-dependent manner. At doses 7.5, 12.5, and 25 mg/kg, 6-beta-naltrexol significantly decreased consumption of a 6% ethanol solution compared with saline control groups. CONCLUSIONS: These data suggest that there may be a potential clinical use for 6-beta-naltrexol in recovering alcoholics. PMID- 11045870 TI - Serotonergic agents and alcoholism treatment: rebirth of the subtype concept--an hypothesis. PMID- 11045871 TI - Molecular basis for treating endometriosis with aromatase inhibitors. AB - Although treatment of one unusually aggressive case of postmenopausal endometriosis with an aromatase inhibitor has been strikingly successful, large clinical trials are required to establish whether aromatase inhibitors will have a significant role in the medical management of endometriosis. Introduction of aromatase inhibitors into the treatment of endometriosis underscores the importance of basic research leading to the development of novel strategies in reproductive disorders. It was shown earlier that aromatase activity was not detectable in normal endometrium. Aromatase, however, is expressed inappropriately in endometriosis and stimulated by prostaglandin E2. Aromatase activity gives rise to local biosynthesis of oestrogen, which, in turn, stimulates prostaglandin E2 production, thus establishing a positive feedback cycle. This favours accumulation of oestrogen and prostaglandins in endometriosis, which is an inflammatory disorder dependent on oestrogen for growth. PMID- 11045872 TI - Prevention of postmenopausal osteoporosis with oestrogen replacement therapy and associated compounds: update on clinical trials since 1995. AB - Hormonal replacement therapy (HRT) is generally regarded as first choice for pharmacological prevention of osteoporosis in women. We reviewed recent studies of HRT regimens and selective oestrogen receptor modulators (SERMs), including controlled trials of at least one-year duration published since 1995 until February 2000 providing data on bone mineral density (BMD) or fractures. Natural and synthetic oestrogens exert a continuum of positive effects on BMD in a dose dependent, though non-proportional, fashion independent of age and mode of administration. Bone loss may be largely prevented by 25 microg transdermal patch oestradiol, 0.3 mg conjugated equine or 0.3 mg esterified oestrogens. Progestogens neither attenuate nor augment the effect of oestrogens; sole use of tibolone prevents bone loss. Both the SERMs, tamoxifen and raloxifene, slightly increase BMD. There are no adequately powered fracture trials for any HRT regimen. Raloxifene 60 mg daily decreases the relative risk of vertebral fractures by at least 30%, as demonstrated by one 3-year fracture study of osteoporotic women. In conclusion, the recommendation to use oestrogen for postmenopausal osteoporosis, given both the lack of fracture trials and the rare trials on long-term use of HRT in (late) postmenopausal women, is not well supported. Fracture trials could overcome shortcomings of the current level of evidence. PMID- 11045873 TI - Weight gain on the combined pill--is it real? AB - Britain has one of the highest teenage pregnancy rates in Western Europe at 8.8 per 1000 live births. Adolescents are very preoccupied with body image and fear weight gain with use of the combined oral contraceptive (COC) pill. Compliance with contraception continues to be a major issue. Is there a real evidence of weight gain? Or are there discrepancies between adolescent perceptions of weight gain with COC use and available scientific evidence? We carried out a comprehensive literature search and did not find evidence for the purported weight gain with use of low dose COCs. Adolescents need reassurance by gynaecologists, general practitioners, family planning doctors and mass media to remove such misperceptions. This will contribute in some way to reduce the high unintended pregnancy rates. PMID- 11045874 TI - Infections in IVF: review and guidelines. AB - Since the inception of in-vitro fertilization (IVF), questions about contamination and the transmission of infection have been raised. In this review, screening for Chlamydia trachomatis, as well as medical and ethical considerations on IVF in couples infected with hepatitis B virus (HBV), hepatitis C virus (HCV) and human immunodeficiency virus (HIV), are discussed. IVF is not contraindicated in case of HBV or HCV infection, but the decision is far more delicate in case of HIV infection. If donor gametes or embryos are used, prevention of infectious disease transmission resides in accurate donor selection, including screening for C. trachomatis, HIV, HBV, cytomegalovirus and Treponema pallidum. In the embryology laboratory, microbial contamination of the IVF system deserves attention, and can be prevented by using sterile technique and supplementing culture media with screened sera or serum substitutes and antibiotics. Persons whose biological material is to be cryopreserved should be screened for HBV, HCV and HIV, and separate containers should be used for infected and non-infected material. Finally, transmission of infectious diseases to laboratory personnel can be prevented by adherence to strict safety guidelines, wearing of protective clothing, HBV vaccination, prohibition of mouth pipetting, and developing a plan for the disposal of bio-hazardous material. PMID- 11045875 TI - Preparation of the cervix for surgical termination of pregnancy in the first trimester. AB - Worldwide, surgical vacuum aspiration is the method of choice of terminating first trimester unwanted pregnancy. Cervical priming prior to surgical evacuation reduces the risks of cervical injury by making the cervix softer and easier to dilate. Over the years, a number of effective methods of cervical priming have became available: osmotic dilators; antiprogesterone and prostaglandins. Of these, prostaglandins remain the most widely used method of cervical preparation. However many of the natural and synthetic analogues of prostaglandins are either expensive or associated with troublesome side-effects. More recently, misoprostol, a synthetic 15-deoxy-16 hydroxy 16-methyl analogue of naturally occurring prostaglandin E, used in the management of peptic ulcers, has established a lead for cervical priming in terms of availability, ease of administration, cost and effectiveness. In fact it appears that both oral and vaginal misoprostol given at dosages of 400 microg are effective for cervical priming when administered 3 h prior to surgical vacuum aspiration. Now that the use of misoprostol for cervical priming has been validated, its widespread use in gynaecological practice is expected. PMID- 11045876 TI - Current status of the cryopreservation of human unfertilized oocytes. AB - Cryopreservation facilitates the long-term storage of oocytes from patients in danger of losing ovarian function and allows greater flexibility in fertility services for other patients. If some of the oocytes collected following ovulation stimulation are stored prior to fertilization, this alleviates many of the ethical concerns associated with embryo preservation. Concerns that cryopreservation could lead to disruption of the spindle and chromosomes, thus leading to genetic abnormalities of the offspring produced, mean that this procedure is not permitted in some countries. The recent spate of human live births from thawed oocytes has prompted the granting of the first licence allowing the use of thawed oocytes in the UK. However, the success rate of this procedure is still low and further research is required to refine these techniques and to develop new ones. PMID- 11045877 TI - Primate and bovine immature oocytes and follicles as sources of fertilizable oocytes. AB - This review discusses the mechanisms underlying ovarian follicle development and the potential of immature follicles and oocytes from non-rodent mammalian species particularly human and bovine to serve as sources of oocytes for the in-vitro production of embryos. Factors that regulate growth and differentiation of unilaminar (primordial and primary) and multilaminar (secondary) follicles and the maturation of oocytes are highlighted. We conclude that many obstacles must still be overcome before fertilizable oocytes can be obtained from human and bovine ovaries, and more research on the quality of and culture conditions for immature oocytes and follicles is required before these can be considered as a source for in-vitro production. PMID- 11045878 TI - Three-dimensional vaginal sonography in obstetrics and gynaecology. AB - Great strides have recently been made in obstetrics and gynaecology secondary to the development of high-performance transvaginal ultrasound (TVS) instruments. However, even this advanced technology can provide only two-dimensional (2D) views of three-dimensional (3D) structures. Although an experienced examiner can easily piece together sequential 2D planes for creating a mental 3D image, individual sectional planes cannot be achieved in a 2D image because of various difficulties. Today, 3D TVS can portray not only individual image planes, it can also store complex tissue volumes which can be digitally manipulated to display a multiplanar view, allowing a systematic tomographic survey of any particular field of interest. The same technology can also display surface rendering and transparency views to provide a more realistic 3D portrayal of various structures and anomalies. The current review provides examples and discussions of the various applications in obstetrics, gynaecology and assisted reproduction in which 3D TVS is a useful supplement to image modality. PMID- 11045879 TI - Villous sprouting: fundamental mechanisms of human placental development. AB - There is increasing evidence that maldevelopment of the placental villous tree can play an important role in the pathogenesis of various pregnancy diseases. In this review we present the most recent advances of cellular and molecular mechanisms involved in the early formation of chorionic villi. In particular we focus our attention on the structural events during early villous sprouting leading to the formation of the mesenchymal villi which are the forerunners of all other villous types, i.e. immature intermediate villi, stem villi, mature intermediate villi and terminal villi. Early villous sprouting starts as 'hot spots' which are circumscribed areas consisting of highly proliferating cytotrophoblastic and stromal cells. The post-proliferative cytotrophoblastic cells fuse with the overlying syncytium leading to the formation of the trophoblastic sprouts. When villous mesenchyme invades the trophoblastic sprouts, the latter are transformed into villous sprouts. The vascularization of the villous sprouts leads to the formation of the mesenchymal villi, the most basic villous type. This process is repeated throughout pregnancy. We analyse the influence of various extracellular matrix molecules, e.g. tenascin and hyaluronic acid, on the formation of hot spots and mesenchymal villi as well as the transformation of the latter in other villous types. We present a critical survey on the data on vessel formation related to villous sprouting and morphogenesis of mesenchymal villi as well as the expression of various angiogenic factors and their receptors. PMID- 11045880 TI - Insulin-like growth factor binding protein-1 (IGFBP-1): a multifunctional role in the human female reproductive tract. AB - Insulin-like growth factor-1 (IGFBP-1) is particularly important in human female reproductive physiology, where it is involved with other factors in a complex system which regulates menstrual cycles, puberty, ovulation, decidualization, implantation and fetal growth. This has implications for clinical obstetrics and gynaecology, where there is evidence for a pathophysiological role for IGFBP-1 in pre-eclampsia, intrauterine growth restriction, polycystic ovarian syndrome and trophoblast and endometrial neoplasms. PMID- 11045881 TI - Direct ovarian effects and safety aspects of GnRH agonists and antagonists. AB - In in-vitro fertilization programmes, gonadotrophin releasing hormone (GnRH) agonists are now routinely used in order to prevent the undesired pre-ovulatory spontaneous luteinizing hormone surge. The first publications are now appearing in which GnRH antagonists are used with the same purpose. More attention should be addressed to the safety aspects of these drugs. This review aims to summarize studies on direct ovarian effects of GnRH agonists and GnRH antagonists in non primates and primates with respect to the functional and morphological aspects in vitro as well as in-vivo. We conclude that there is a wide variety of functional and morphological effects of GnRH analogues on the ovary. The sometimes paradoxical effects indicate that a variety of factors may be involved in the various processes. Those factors are: (i) the type and dose of the analogue, (ii) the different regimens of administration, (iii) ovarian status at the time of exposure, (iv) ovarian cell types in in-vitro systems, (v) hormonal pre-treatment of these cultures, (vi) the type of hormonal stimulation added to the in-vitro culture, (vii) further methodological differences in the experiments and finally (viii) physiological variations in GnRH receptor abundance which depends on species and/or timing in the cycle. With the increasing number of patients using GnRH analogues in assisted reproduction treatments, there will be an increasing number of pregnancies exposed to these drugs. So far, there does not appear to be an increased risk of birth defects or pregnancy wastage in human pregnancies exposed to daily low-dose GnRH agonist therapy in the first weeks of gestation. PMID- 11045882 TI - The regulation of the human corpus luteum steroidogenesis: a hypothesis? AB - The corpus luteum (CL) is an important endocrine organ in the menstrual cycle and in pregnancy. The regulation of its hormonal production has been extensively studied. The steroidogenic abilities of the CL can be rescued by human chorionic gonadotrophin (HCG) but its role in the maintenance of CL function is not clear. We will discuss the hypothesis that there are fetoplacental factors, other than HCG, that modulate CL steroidogenesis. PMID- 11045883 TI - Influence of nanoparticles on the brain-to-serum distribution and the metabolism of valproic acid in mice. AB - The suitability of nanoparticles as a drug-carrier system for the antiepileptic valproic acid has been studied in mice. The aim of the study was to increase the brain-to-serum ratio of the drug to reduce dose-related side effects in the periphery. The influence of nanoparticles on the metabolism of valproic acid was also investigated. The serum kinetics and the brain tissue levels of valproic acid were not altered by administration with nanoparticles. However, the nanoparticles did inhibit the metabolic degradation of valproic acid via mitochondrial beta-oxidation but did not influence any other metabolic pathway. It can be concluded that nanoparticles loaded with valproic acid may help to reduce the toxic side effects of valproate therapy, not by reducing the therapeutically necessary dosage but by inhibition of formation of toxic metabolites. Using their ability to selectively block a pathway nanoparticles may serve as a tool to investigate the metabolic origin of metabolites and their contribution to therapeutic efficacy and side effects. PMID- 11045884 TI - Ileal uptake of polyalkylcyanoacrylate nanocapsules in the rat. AB - The ileal uptake of polyalkylcyanoacrylate nanocapsules (less than 300 nm in diameter) has been investigated in the rat. Iodised oil (Lipiodol) was used as the tracer for X-ray microprobe analysis in scanning electron microscopy. Lipiodol nanocapsules, or an emulsion of Lipiodol, were administered in the lumen of an isolated ileal loop of rat. Lipiodol nanocapsules improved the absorption of the tracer as indicated by increased concentrations of iodine in the mesenteric blood (+27%, P < 0-01, compared with Lipiodol emulsion). Intestinal biopsies were taken at different time points and the samples underwent cryofixation and freeze-drying. The nanocapsules were characterized by their strong iodine emission, and electron microscopy of the biopsy samples revealed nanocapsules in the intraluminal mucus of the non-follicular epithelium, then in the intercellular spaces between enterocytes, and finally the nanocapsules were found within intravillus capillaries. However, nanocapsules were most abundant in the Peyer's patches, where the intestinal epithelium had been crossed by way of the specialized epithelial cells, designated membranous cells, or M cells, and their adjacent absorptive cells. These observations were confirmed quantitatively by measuring iodine concentrations in the various tissue compartments. Ten minutes after the intraluminal administration of Lipiodol nanocapsules, the emission of iodine peaked in the mucus (+77%, P < 0.01), in M cells (+366%, P <0.001), in enterocytes adjacent to M cells (+70%, P < 0.05) and in lymph vessels (+59%, P < 0.05). Polyalkylcyanoacrylate nanocapsules were able to pass through the ileal mucosa of the rat via a paracellular pathway in the non-follicular epithelium, and most predominantly, via M cells and adjacent enterocytes in Peyer's patches. PMID- 11045885 TI - Biodistribution of stealth and non-stealth solid lipid nanospheres after intravenous administration to rats. AB - Drug-free stealth and non-stealth solid lipid nanospheres (SLNs) were administered intravenously to rats to evaluate their tissue distribution and their transport across the blood-brain barrier. Two types of experiments were performed using unlabelled and labelled SLNs. Rats were administered labelled non stealth or stealth nanospheres (NSSLNs and SSLNs) and their tissue distribution was monitored for 60 min. In another experiment, rats were injected with unlabelled NSSLNs or SSLNs and the cerebrospinal fluid (CSF) was analysed using transmission electron microscopy (TEM) to confirm the presence of the SLNs. Some differences were found in the biodistribution between labelled NSSLNs and SSLNs. In particular, the radioactivity in the liver and the lung was much lower for SSLNs than for NSSLNs, confirming a difference in their uptake. Both types of SLNs were detected in the brain. TEM analysis showed both types of SLNs in rat CSF. PMID- 11045886 TI - Clearance of fentanyl, alfentanil, methohexitone, thiopentone and ketamine in relation to estimated hepatic blood flow in several animal species: application to prediction of clearance in man. AB - We have used estimated hepatic blood flow (Qhep) as an aid to evaluate clearance (CL) values in animals and to predict clearance in man of five anaesthetic agents: fentanyl, alfentanil, methohexitone, thiopentone and ketamine. The disposition of methohexitone was determined in rats and that of ketamine in rats, rabbits and pigs. Further data were compiled from the literature and supplemented experimentally as needed. Allometric interspecies scaling, according to three different methods, was used to estimate blood clearance and unbound clearance (CLu) in man. The results of scaling according to the three different methods were evaluated in relation to estimated hepatic extraction ratio (CL/Qhep) of the drugs. In most animals the clearance of the drugs were comparable with or lower than estimated Qhep. However, ketamine showed extensive extrahepatic clearance in rabbits. Prediction of clearance in man was successful by at least one method for all five drugs, while prediction of CLu generally failed. Estimates of CL/Qhep gave no indication as to the choice of the best method. Volume of distribution at steady state could be predicted for alfentanil, thiopentone and ketamine. Comparison of clearance with Qhep should be used to evaluate clearance data in animals, however estimation of hepatic extraction ratios appears to be of little use for allometric scaling. The use of ketamine as an anaesthetic agent in rabbits is questionable, while the use of fentanyl in pigs, methohexitone in rats and ketamine in rats and pigs is well supported by the pharmacokinetic data. PMID- 11045887 TI - Comparative pharmacodynamics and pharmacokinetics of candesartan and losartan in man. AB - The angiotensin II antagonistic effects of candesartan and losartan were compared in-vivo after single and repeated doses. Effects were related to antagonistic activity in plasma. In this double-blind, crossover study, 12 healthy male volunteers received, in random order, daily oral doses of 8 mg candesartan cilexetil or 50 mg losartan for seven days. On day 1 and day 8, dynamics and kinetics were assessed up to 48 h after dosing. Antagonistic effect was determined from the antagonist-induced rightward shifts of the diastolic blood pressure response curves to exogenously administered angiotensin II measured as the dose ratio (DR). The antagonistic activity in plasma was measured using an ex vivo/in-vitro radioreceptor assay. Specific high-performance liquid chromatography assays determined plasma concentrations of candesartan, losartan and its active metabolite EXP-3174. The pharmacokinetic properties of candesartan and losartan were comparable and antagonistic activity in plasma almost identical (ratio candesartan: losartan = 0.97 and 1-2 after single and multiple doses, respectively). However, the antagonistic effects of candesartan and losartan in vivo were quite different. Twenty-four hours after single dosing with candesartan a clinically relevant rightward shift in the angiotensin II dose-response curve (DR= 3.2) occurred that was more pronounced than that following losartan administration (DR=2.1, ratio candesartan: losartan= 1.65). Twenty-four hours after multiple doses of candesartan or losartan, the values of the DR were 4.8 and 2.3, respectively (ratio candesartan: losartan = 1.94). The values of DR for candesartan were significantly higher compared with losartan between 6 and 36h after a single dose and between 3 and 24 h post-dose following multiple dose administration. A counter-clockwise hysteresis was apparent between antagonistic activity in plasma and antagonistic effect. Despite equivalent angiotensin II antagonistic activity in plasma, the pharmacodynamic effect of candesartan cilexetil was greater than that of losartan. Candesartan appeared to have a slower off-rate from the angiotensin AT1-receptor compared with losartan, nevertheless differences in distributional phenomena or the extent of insurmountable antagonistic activity cannot be ruled out. PMID- 11045888 TI - Chronopharmacokinetics of sumatriptan in healthy human subjects. AB - Rhythms in the onset and symptoms of several diseases are well established. Migraine is a disorder that exhibits periodicity in its symptoms and so chronotherapy may be beneficial in treating the problem. Designing a chronotherapeutic schedule requires chronopharmacokinetic and chronopharmacodynamic data of the drugs prescribed. We have studied the chronopharmacokinetics of sumatriptan, a drug of choice in migraine treatment. Twelve healthy male volunteers were treated with 100 mg sumatriptan orally at 0700, 1300, 1900 and 0100 h in a randomized 4 x 4 Latin square crossover design, with a wash-out period of one week. Serum samples were analysed by high performance liquid chromatography with an electrochemical detector. Pharmacokinetic parameters were calculated using noncompartmental methods. The pharmacokinetic parameters were analysed using analysis of variance and a two tailed paired t-test at the probability of 95%. The mean peak serum concentration following the 0700 h administration (Cmax; 59.09 +/- 10.53 ng mL(-1)) was significantly (P < 0.05; n = 12) higher than after the 1900 h administration (Cmax 41.88 +/- 12.21 ngmL(-1)). The mean area under the serum concentration-time curve from time zero to the last time-point (AUCo-t), the area under the serum concentration-time curve from zero to infinity (AUC 0-infinity), and the area under the first moment curve (AUMC) were significantly (P < 0.05; n = 12) higher following the 0700 and 0100 h administrations than after the 1900 h administration. Following administration at 0700 h, the mean oral clearance (CLs/f; 781 +/- 186 mL h(- 1) kg(-1)) and the apparent volume of distribution (Vd/f; 2,379 +/- 684) were significantly lower (P < 0.05; n = 12) than after the 1900 h administration (CLs/f 1,208 +/- 458 mL h(-1) kg(-1), Vd/f 4,655 +/- 2,096 mL kg(-1)). The mean Vd/f value was again lower after the 1300h administration than after the 1900 h administration (2,763 +/- 1,417 vs 4,655 +/- 2,096 mL kg( 1); P < 0.05; n = 12). The variations may be due to the time dependent changes in the extent of absorption and/or circadian variations in hepatic blood flow. PMID- 11045889 TI - The kinetic profile of gentamicin in premature neonates. AB - The kinetic profile of gentamicin in premature infants has been studied to enable the development of optimized dosage schedules for neonatal intensive-care units and to stress the relationship between the pharmacokinetic parameters and several demographic, developmental and clinical factors which might be associated with changes in gentamicin disposition. Sixty-eight newborn patients of 24- to 34 weeks gestational age and 600-3,100 g current weight in their first week of life, undergoing routine therapeutic drug monitoring of their gentamicin serum levels, were included in this retrospective analysis. Gentamicin pharmacokinetic parameters were determined through non-linear regression by using a single compartment open model. By regression analysis the current weight (g) was shown to be the strongest co-variate, and both gentamicin clearance (L h(-1)) and volume of distribution (L) had to be normalized. Additionally, gentamicin clearance depended on gestational age with a cut-off at 30 weeks, which allowed the division of the overall population into two subsets (< 30 weeks and between 30-34 weeks of gestational age). The younger neonates (<30 weeks of gestational age) showed a lower gentamicin clearance (0.0288 vs 0.0340 L h(-1) kg(-1)), a slightly higher volume of distribution (0.464 vs 0.435 L kg(-1)), and a longer half-life (11.17 vs 8.88 h) compared with the older subgroup (30-34 weeks of gestational age). On the basis of the pharmacokinetic parameters obtained, we suggest loading doses of 3.7 and 3.5 mg kg(-1) for the two subgroups of neonates (<30 weeks and 30-34 weeks of gestational age), respectively. The appropriate maintenance doses in accordance with the characteristics of the patients should be 2.8 mgkg(-1)/24h and 2.6 mg kg(-1)/18 h for neonates < 30 weeks and between 30 34 weeks of gestational age, respectively. Finally, when compared with previous studies, the information obtained on the pharmacokinetics and determinants of the pharmacokinetic variability of gentamicin in neonates was shown to be consistent. PMID- 11045890 TI - Pharmacokinetics and hepatoprotective effects of 2-methylaminoethyl-4,4' dimethoxy-5 ,6,5',6'dimethylenedioxybiphenyl-2-carboxylic acid-2'-carboxylate monohydrochloride in rats with CCl4-induced acute hepatic failure. AB - The pharmacokinetics and hepatoprotective effects of 2-methylaminoethyl-4,4' dimethoxy-5,6,5',6'-dimethylenedioxybip henyl-2-carboxylic acid-2'-carboxylate monohydrochloride (DDB-S) have been investigated in rats with CCl4-induced acute hepatic failure. To study the pharmacokinetics of DDB-S, rats were divided into a control group and a CCl4-intoxicated group. DDB-S 50 mg kg(-1) was administered by intravenous bolus injection to both groups of rats. In the CCl4-intoxicated rats the plasma concentrations of DDB-S were significantly higher, the area under the plasma concentration-time curve from time zero to time infinity was significantly greater (6-46 vs 3.34 mg min mL(-1)), and the total body (7.74 vs 15.0 mL min(-1) kg(-1)), renal (2.55 vs 5.10 mL min(-1) kg(-1)), nonrenal (5.07 vs 9.65 mL min(-1) kg(-1)), and biliary (1.48 vs 2.69 mL min(-1) kg(-1)) clearances were significantly slower compared with the control rats. This could be due to decreased hepatic cytochrome P450 activity and impaired kidney function induced by CCl4. To study the hepatoprotective effects of DDB-S, rats were divided into three groups, control rats and CCl4-intoxicated rats with or without DDB-S pretreatment (50 mg kg(-1) i.p.). The effects of DDB-S pretreatment on CCl4 induced liver injury were considerable; the serum levels of alanine transaminase, aspartate transaminase, and alkaline phosphatase were significantly lower by 54.3, 44.6 and 67.2%, respectively, compared with the CCl4-intoxicated-only group. In an in-vitro study, rat hepatocytes were exposed to fresh medium containing 10 mM CCl4 and various concentrations of DDB-S (10 or 100 microg mL( 1)). The levels of alanine transaminase and aspartate transaminase in the medium were measured as an indicator of hepatocyte injury. DDB-S dose-dependently decreased the levels of alanine transaminase and aspartate transaminase compared with CCl4-intoxication only. These results indicate that DDB-S has hepatoprotective activity. PMID- 11045891 TI - Analogues of arginine vasopressin modified in position 2 or 3 with naphthylalanine: selective antagonists of oxytocin in-vitro. AB - In this study we describe the synthesis and some pharmacological properties of six new analogues of arginine vasopressin (AVP). Five of the peptides were substituted in position 2 with L-1-naphthylalanine (L-1-Nal) or D-1 naphthylalanine (D-1-Nal); one had D-1-Nal in position 3. All analogues were tested in bioassays for pressor and antidiuretic activity. We also tested the uterotonic activity of the peptides in-vitro. Two of the new peptides were moderately potent V1a and oxytocin antagonists. The modifications proposed resulted in a drop or the removal of antidiuretic activity and in the removal of pressor activity, or conversion into moderate antagonists. Two peptides ([Mpa1, (L-1-Nal)2]AVP and [Mpal, (D-1-Nal)2]AVP) which appear not to interact with V1a and V2 receptors were exceptionally selective oxytocin antagonists in vitro. These compounds with selective oxytocin antagonistic activity may be promising candidates for the development of potential tocolytic agents for the prevention of pre-term labour. PMID- 11045892 TI - In-vitro hydrolysis, permeability, and ocular uptake of prodrugs of N-[4 (benzoylamino)phenylsulfonyl]glycine, a novel aldose reductase inhibitor. AB - To enhance the ocular uptake of N-[4-(benzoylamino)phenylsulfonyl]glycine (BAPSG), two ester (methyl and isopropyl) prodrugs were synthesized and evaluated for their stability in various buffers (pH 1-9), hydrolysis in rabbit ocular tissues (cornea, conjunctiva, iris-ciliary body, lens, aqueous humor, and vitreous humor), transport across cornea and conjunctiva, and in-vivo uptake following topical administration. Over the pH range of 1-9, the rate constants for degradation ranged from 5.67 to 218.9 x 10(-3) h(-1) for the methyl ester and from 3.14 to 4.45 x 10(-3) h(-1) for the isopropyl ester. At all pH conditions, the isopropyl ester was more stable when compared with the methyl ester. A change in buffer concentration at pH 7.4 did not influence the stability of the prodrugs. The prodrugs were rapidly hydrolysed in the tissue homogenates, with the rate constants for hydrolysis ranging from 1.98 to 7.2x 10(-3) min(-1) for the methyl ester and 3.32 to 6.53 x 10(-3) min(-1) for the isopropyl ester. The in-vitro permeability of the methyl ester was less than the parent drug across cornea and conjunctiva. Isopropyl ester levels were not detectable in the receiver chamber even at the end of the 4-h transport study. Following topical administration of BAPSG and the two prodrugs at a dose of 60 microg/eye, the lowest levels were seen in vitreous humor for parent compound and its methyl ester. In general, the tissue uptake of methyl ester was less than BAPSG. Isopropyl ester levels were below detection limits in all the ocular tissues. Lipophilic ester prodrugs of BAPSG showed good aqueous solution stability in tissue homogenates. However, these prodrugs lacking the free carboxylate anion exhibited reduced in-vitro permeability and in-vivo uptake, suggesting the importance of free carboxylate anion in the delivery of BAPSG. PMID- 11045893 TI - Structure-activity relationships for the inhibition of lipid peroxidation and the scavenging of free radicals by synthetic symmetrical curcumin analogues. AB - A number of ring substituted analogues of curcumin were synthesized. Their antioxidant properties were studied using three models, inhibition of lipid peroxidation, scavenging of 1,1'-diphenyl-2-picrylhydrazyl (DPPH) and 2,2' azinobis(3-ethyl-benzthiazoline-6-sulphonate radical (ABTS+.). In all the models, the phenolic analogues were more active than the non-phenolic analogues, some of which were inactive. The highest antioxidant activity was obtained when the phenolic group was sterically hindered by the introduction of two methyl groups at the ortho position. This and several other compounds were more active than the standard antioxidants alpha-tocopherol and trolox. This study has demonstrated that the phenolic group is important for the antioxidant activity of curcumin and that the structural features that enhance the antioxidant properties of phenols are optimized in curcumin to a significant extent. PMID- 11045894 TI - Comparative pharmacological study of ropinirole (SKF-101468) and its metabolites in rats. AB - The dopamine receptor agonist ropinirole (SKF-101468) is used to treat Parkinson's disease. Ropinirole is metabolized by two routes to a series of different metabolites although the predominant pathway is species-dependent. It is unknown whether any of the metabolites contribute to its antiparkinsonian activity and whether D3 or D2 receptor agonist activity plays a preferential role. Therefore ropinirole and its primary metabolites, SKF-104557, SKF-97930 and SKF-96990, and the rat metabolite, SKF-89124 were tested in the 6-hydroxydopamine lesion model of Parkinson's disease. SKF-89124 and SKF-96990 were also assayed in radioligand binding and microphysiometer functional assays at cloned human dopamine D2 and D3. Ropinirole and SKF-89124 were equipotent in-vivo, and produced dose-related increases in circling at 0.05-0.8 mg kg(-1), s.c. (ropinirole) and 0.05-0.75 mg kg(-1), s.c. (SKF-89124). Neither SKF-96990 or SKF 97930, at doses up to 15 mg kg(-1), increased the circling rate. Some circling was observed with 15 mg kg(-1) SKF-104557 but the response was less than half that produced by ropinirole (0.8 mgkg(-1)). SKF-104557 was 150-fold less potent than ropinirole. SKF-89124 possessed-30-fold higher affinity for D3 over D2 receptors in radioligand binding studies, but was not selective in the functional microphysiometer assay. SKF-96990 was 10-fold selective for D3 over D2 receptors in the radioligand binding assay. Ropinirole and SKF-104557 are 20-fold selective for D3 over D2 receptors in radioligand binding assays whereas in microphysiometry, selectivity is 10-fold. SKF-97930 is inactive in radioligand binding and microphysiometer assays. Primary metabolites of ropinirole did not contribute significantly to its activity in this model of Parkinson's disease. The lack of dopamine D3/D2 receptor selectivity for ropinirole rules out the possibility of attributing the degree of either D2 or D3 receptor activity to the behavioural efficacy of ropinirole. PMID- 11045895 TI - Antitussive efficacy of dextromethorphan in cough associated with acute upper respiratory tract infection. AB - Dextromethorphan is one of the most widely used antitussives for the treatment of cough associated with acute upper respiratory tract infection. However, there is very little data to support the efficacy of dextromethorphan in this disease state. This aim of this study was to obtain more information about the efficacy of a single dose of 30 mg dextromethorphan in the treatment of cough associated with acute upper respiratory tract infection. The study was a double-blind, stratified, randomized and parallel group design. Both objective and subjective measurements of cough were recorded over 10-min recording periods in a quiet room before (baseline) and at 90, 135 and 180 min after treatment. Forty-three patients (30 females and 13 males), mean age 22.9 years (range 18-46 years), with acute dry or slightly productive cough and otherwise healthy were included in the study. Patients were randomized to placebo treatment (n = 22) and dextromethorphan treatment (n=21). The results showed similar trends in both treatment groups with statistically significant reductions (P < 0.05) in cough sound pressure level (CSPL), cough frequency (CF) and subjective scores for cough severity within treatment groups but little difference between the treatment groups during the study period. The only statistically significant difference between treatment groups was for the mean CSPL changes from baseline to 90 min (P=0.019). There was a significant positive correlation between CSPL and CF (r = 0.752, P= 0.000) for changes in cough measurements from baseline to 90 min after treatment and this indicates that CSPL may be a useful measure of cough severity. This study provides very little if any support for clinically significant antitussive activity of a single 30 mg dose of dextromethorphan in patients with cough associated with acute upper respiratory tract infection. PMID- 11045896 TI - Comparative cerebrospinal fluid diffusion of imipenem and meropenem in rats. AB - The main objective of this study was to compare the cerebrospinal fluid (CSF) diffusion of imipenem and meropenem at steady state, following intravenous infusions at various rates in rats. A preliminary experiment was conducted to estimate the elimination half-lives of these two carbapenem antibiotics, and then to evaluate the infusion duration necessary to reach steady state. CSF diffusion of imipenem was essentially linear over the wide range of infusion rates (66 1,320microg min(-1)) and corresponding steady-state plasma concentrations (11.7 443.0 microg mL(-1)). Conversely the CSF diffusion of meropenem was saturable, with a predicted maximum CSF concentration equal to 1.3 microg mL(-1). Extrapolation of these data to the clinical situation may not be possible since the rats had normal blood-brain and blood-CSF barriers whereas patients with diseases such as meningitis may not. However, it is suggested that the observed differences in the diffusion characteristics of imipenem and meropenem may be partly responsible for their differences in toxicity and efficacy at the central level. PMID- 11045897 TI - A post-ischaemic single administration of galanthamine, a cholinesterase inhibitor, improves learning ability in rats. AB - Transient forebrain ischaemia is widely observed in clinical practice. We have examined the effect of a single administration of the cholinesterase inhibitor galanthamine (2mg kg(-1) i.p.) 25 min after reperfusion in male Sprague-Dawley rats (180 +/- 20 g) after a 20-min common carotid artery occlusion. Twenty-four hours post-ischaemia there was no difference in motor co-ordination or muscle tonus of the rats treated with or without galanthamine as assessed by the rota rod test. Learning ability was examined using the shuttle-box test, evaluating the latency time and the number of errors for six days in succession. The performance of the ischaemic saline-injected rats was significantly impaired on days 4, 5, 6 (latency time) compared with the non-ischaemic rats and with the ischaemic animals administered galanthamine (P < 0.05). Similar results were obtained when counting the number of errors (failure to cross the cage during conditioned or unconditioned stimulus). The monitoring of body temperature during the first 12-h post-ischaemia did not show any significant difference between the groups. The data showed a beneficial effect of galanthamine on the recovery of learning ability when administered once only post-ischaemia. This suggests a direct effect on the early pathologic mechanisms of CNS damage. Cholinesterase inhibitors may prove useful in the early clinical treatment of ischaemic conditions. PMID- 11045898 TI - The anti-ulcerogenic effect of an alkaloidal fraction from Mikania cordata on diclofenac sodium-induced gastrointestinal lesions in rats. AB - A decoction of Mikania cordata (Compositae) is commonly used for the treatment of gastric ulcer in the Rajbari district of Bangladesh. We have evaluated the anti ulcerogenic effect of the alkaloidal fraction from the whole plant of M. cordata on diclofenac sodium-induced gastrointestinal lesion in rats. Long Evan's rats were divided into five groups. The control group was kept undisturbed. The vehicle group received vehicle after a 48-h fast. The diclofenac group received diclofenac sodium suspension (80 mg kg(-1)) after a 48-h fast. The diclofenac ranitidine group (anti-ulcer drug used as a standard) received 35 mg kg(-1) ranitidine hydrochloride suspension 1 h after diclofenac sodium administration, after a 48-h fast. The diclofenac-extract group received alkaloidal fraction (50 mg kg(-1)) 1 h after diclofenac administration, after a 48-h fast. The biochemical, morphological and histological changes were studied. The data showed that the pH values of the stomach and duodenum were increased significantly (P < 0.001) in the alkaloidal-administered group compared with the control group (3.09 +/- 0.0429 vs 2.07 +/- 0.0339 and 6.79 +/- 0.1162 vs 6.19 +/- 0.1273, respectively). There were significant changes (P < 0.001) detected in the morphological study. The ulcer index of the stomach (0.268 +/- 0.0346) and of the duodenum (0.050 +/- 0.0129) in the alkaloidal-administered group were significantly lower than the diclofenac-only administered group (0.691 +/- 0.0184 and 0.0933 +/- 0.0138, respectively). According to the grading of tissue damage in the histological study, there were less or no lesions on the gastrointestinal mucosa of the alkaloidal-administered group compared with the diclofenac-only group (0 vs 5, respectively). When the results of the alkaloid extract group where compared with the ranitidine hydrochloride group a similar or more potent effect was observed with the alkaloidal extract group. The results of this study revealed that the bioactive principles of M. cordata have anti-ulcerogenic effects. The results validate the traditional use of this plant for the treatment of gastric ulcer in Bangladesh. PMID- 11045899 TI - Antioxidant lignans from Machilus thunbergii protect CCl4-injured primary cultures of rat hepatocytes. AB - Eleven lignans (1-11) were isolated from the CH2Cl2 fraction of the bark of Machilus thunbergii Sieb. et Zucc. (Lauraceae). These were identified as (-) acuminatin (1), (-)-isoguaiacin (2), meso-dihydroguaiaretic acid (3), (+) galbacin (4), (-)-sesamin (5), (+)-galbelgin (6), machilin A (7), machilin G (8), licarin A (9), and nectandrin A (10) and B (11). Primary cultures of rat hepatocytes were co-incubated for 90 min with the hepatotoxin CCl4 and each of the 11 lignans (50 microM). Hepatoprotective activity was determined by measuring the level of glutamic pyruvic transaminase released into the medium from the primary cultures of rat hepatocytes. (-)-Acuminatin, (-)-isoguaiacin and meso dihydroguaiaretic acid all significantly reduced the level of glutamic pyruvic transaminase released. Further investigation revealed that these three compounds significantly preserved the levels and the activities of glutathione, superoxide dismutase, glutathione peroxidase and catalase. (-)-Acuminatin, (-)-isoguaiacin and meso-dihydroguaiaretic acid also ameliorated lipid peroxidation as demonstrated by a reduction of malondialdehyde production. These results suggest that (-)-acuminatin, (-)-isoguaiacin and meso-dihydroguaiaretic acid exert diverse hepatoprotective activities, perhaps by serving as potent antioxidants. PMID- 11045900 TI - Orphan G-protein coupled receptors: novel drug targets for the pharmaceutical industry. PMID- 11045901 TI - Conformational analysis of 2-[2-(3-methoxyphenyl) ethyl]phenoxyalkylamines with high 5-HT2 receptor binding affinity. AB - A conformational analysis of three groups of 2-[2-(3 methoxyphenyl)ethyl]phenoxyalkylamines with high 5-HT2 receptor binding affinity has been performed using the systematic search. Two groups of compounds with different lengths of alkyl chains connecting the amine nitrogen and the central oxygen showed a one order difference in their 5-HT2 receptor binding affinity. The computational analysis of these compounds confirmed the differences in the N- O distances between the two groups, quantitatively. A probable active conformation was proposed based on a superimposition of the stable conformations over a rigid molecule, mianserin. Two hydroxy derivatives in the third group showed a significant difference in their binding affinity depending on the stereochemistry of the hydroxy group. The difference in the energetically favorable order of the stable conformations reasonably explained the relationship between the stereochemistry and the binding activity. A molecular dynamics-based conformational search was also carried out to compare it with the systematic search. PMID- 11045902 TI - Pharmacophore/receptor models for GABA(A)/BzR alpha2beta3gamma2, alpha3beta3gamma2 and alpha4beta3gamma2 recombinant subtypes. Included volume analysis and comparison to alpha1beta3gamma2, alpha5beta3gamma2, and alpha6beta3gamma2 subtypes. AB - Pharmacophore/receptor models for 6 recombinant GABA(A)/BzR subtypes (alphax beta3gamma2, x = 1-6) have been established via an SAR ligand mapping approach. This study was based on the affinities of 166 BzR ligands at 6 distinct (alpha1 6beta3gamma2) recombinant GABA(A)/BzR receptor subtypes from at least twelve different structural families. Examination of the included volumes indicated that the shapes of binding pockets for alpha1, alpha2 and alpha3 subtypes are very similar to each other. Region L2 for the alpha5 containing subtype appeared to be larger in size than the analogous region of the other receptor subtypes. Region L(Di), in contrast, appeared to be larger in the alpha1 subtype than in the other subtypes. Moreover, region L3 in the alpha6 subtype is either very small or nonexistent in this diazepam insensitive "DI" subtype as compared to the other subtypes. Preliminary results for the alpha4-containing receptor subtype (DI) indicate that L3 in the alpha4 subtype suffers a similar fate. Use of the pharmacophore/receptor models for these subtypes have resulted in the design of novel BzR ligands selective for the alpha5beta3gamma2, receptor subtype. PMID- 11045903 TI - Synthesis and binding studies on a new series of arylpiperazino benzazol-2-one and benzoxazin-3-one derivatives as selective D4 ligands. AB - A series of new arylpiperazinomethyl derivatives was designed and studied as potential D4 ligands. The synthesis of these compounds required an original synthetic route. Some of the tested compounds were found to be as potent as clozapine at D4 receptors. Moreover, compounds which displayed a high D2/D4 selectivity ratio (>122) were selected for further pharmacological evaluation. PMID- 11045904 TI - Hansch analysis of antimalarial cyclic peroxy ketals with physicochemical and electrotopological parameters. AB - Hansch analysis of some antimalarial cyclic peroxy ketals (IV) having structural variations at the para substituted phenyl ring and an alicyclic ring of different size reveals that electronic and steric parameters of the phenyl ring substituents are important for explaining the variation in the activity while hydrophobicity parameter is of little significance. Electron withdrawing substituents with higher MR (molar refractivity) or V(W) (van der Waals volume) are preferred for the activity. Use of structural descriptors suggests that presence of a seven membered alicyclic ring attached to the peroxy bridge containing ring is conducive to the activity. Application of electrotopological state atom index (ETSAI) suggests a pharmacophore containing the peroxy bridge. This is corroborated by earlier observation on importance of oxygen atoms of the peroxy linkage of artemisinin for antimalarial activity. Although incorporation of ETSAI into Hansch model does not improve the relations, the electronic parameter sigma is found to be significantly correlated with it. PMID- 11045905 TI - Magnetic stimulation of the cavernous nerve for the treatment of erectile dysfunction in humans. AB - A recent study in dogs has demonstrated that magnetic stimulation (MS) of the cavernous nerve produced an increase of the intracorporeal pressure and full penile erection. In view of these results, we tested the possible application of this procedure in humans with erectile dysfunction (ED). The study comprised 32 patients with ED (age 38.3 +/- 9.6 y) and 20 healthy volunteers (age 36.8 +/- 8.8 y). Routine erectile function tests suggested that impotence was neurogenic. A magnetic coil was placed over the dorsal aspect of the penis in the vicinity of the symphysis pubis. MS was performed using a stimulation of 40% intensity, 20 Hz frequency, 50 s on and 50 s off for 10 minutes duration. In the healthy volunteers, the coil was placed as aforementioned but was not activated. The intracorporeal pressure was recorded and penile tumescence and rigidity observed during MS in the patients and without stimulation in the controls. MS led to gradual increase in length and diameter of the penis until full erection was achieved; the penis became firm, rigid and pulsatile. The intracorporeal pressure increased significantly (P < 0.0001) at full erection. Mean latency to full erection was 19.3 +/- 3.4 s. Upon off-stimulation, penile erection and intracorporeal pressure returned to baseline after a mean of 22.7 +/- 3.2 s. Penile and pressure response to MS was resumed after an off-time of 50 s. The response was reproducible infinitely if the off-time was observed. The controls showed no penile tumescence or rigidity or increase of the intracorporeal pressure. In conclusion, MS of the cavernous nerve is effective in inducing penile rigidity. It is a simple, easy and non-invasive method which has no adverse effects. It might prove to be suitable for application in patients with ED. PMID- 11045906 TI - Changes in sexual behavior after orthopedic replacement of hip or knee in elderly males--a prospective study. AB - The aim of this study was to investigate two questions: Does arthroplastic surgery affect the patient's status as being sexually active; and if patients are sexually active, does surgery affect their erectile function? The study was designed prospectively and the patients filled in a questionnaire concerning sexuality and erectile function before and 6 months after alloplastic hip-or knee surgery. Ninety-nine males were included, mean age 70.6 y. The results demonstrate that 17% of patients lost a sexual activity that they had preoperatively, and no one regained sexual activity after surgery. A correlation between increasing age and risk of losing sexual activity was demonstrated. 26.1% lost a normal erectile function they had preoperatively, while 6.7% regained normal erections. A similar correlation between increasing age and increased risk was demonstrated. From this study of elderly males undergoing orthopaedic alloplastic surgery it is concluded that the risk of losing sexual activity and erectile capability is increased after surgery, and especially in the group where sexual functions are already impaired. PMID- 11045907 TI - A surgical algorithm for penile prosthesis placement in men with erectile failure and Peyronie's disease. AB - We developed an algorithm for surgical management and placement of penile prostheses in patients with erectile failure (ED) and Peyronie's disease (PD). We identified 46 men ages 40 to 77 y with PD who could not attain an adequate erection with sexual stimulation and pharmacotherapy. All men were candidates for penile straightening and inflatable prosthesis placement using the following algorithm. Manual molding was attempted initially, followed by tunica incision for insufficient straightening. For tunical defects greater than 2 cm, polytetrafluoroethylene (PTFE) patch grafting was performed to prevent prosthesis cylinder herniation and recurrent deformity from cicatrix contraction. Mean preoperative penile curvature was 53 degrees (0-90). Prosthesis implantation with manual molding, implant with plaque incision, and implant with plaque incision and PTFE grafting were successfully accomplished in 25 (54%), 12 (26%), and nine (20)% respectively. Mean follow-up was 39 months (range 1-74). Full erectile capacity with a straight phallus was achieved in all patients. Complications included temporary (< 8 months) decreased penile sensation in four (9%), mild (< 2 cm) penile shortening in three (7%), delayed ejaculation in one (2%), and infection requiring explanation in one diabetic male (2%). All of the implanted prostheses provided satisfactory rigidity with no mechanical failures or recurrent curvature. We conclude that inflatable penile prosthesis implantation is a safe and effective therapy with a high satisfaction rate in men with ED and PD. The developed algorithm helps define prosthesis placement and straightening techniques to obtain optimal results with minimal complications. PMID- 11045908 TI - Corporeal counter incisions: a simplified approach to penile prosthesis implantation in fibrotic cases. AB - OBJECTIVE: To evaluate the effectiveness of the technique of corporeal counter incisions for penile prosthesis implantation in cases with marked corporeal fibrosis. METHODS: Creating subtunical tunnels that are stretched transversely by the dilamezinsert instrument & Heggars dilators with its tip entering from one side and protruding from the counter incision to avoid urethral or corporeal perforation. PATIENTS: Seventeen patients underwent this technique, mean age 56 y (range 42-71 y); 12 post removal of an infected prosthesis and five post priapism. RESULTS: In all cases, two rods were successfully implanted. In one case, a crural perforation occurred that was repaired intra-operatively. CONCLUSION: Corporeal counter incisions with transverse tissue stretching is a relatively simple technique to implant semirigid implants in fibrosed corpora. PMID- 11045909 TI - Soluble guanylate cyclase and cGMP-dependent protein kinase I expression in the human corpus cavernosum. AB - Nitric oxide (NO) as a mediator in smooth muscle cells causes rapid and robust increases in cGMP levels. The cGMP-dependent protein kinase I has emerged as an important signal transduction mediator for smooth muscle relaxation. The purpose of this study was to examine the existence and distribution of two key enzymes of the NO/cGMP pathway, the cGMP-dependent kinase I (cGK I) and the soluble guanylate cyclase (sGC) in human cavernosal tissue. The expression of the enzymes were examined in corpus cavernosum specimens of 23 patients. Eleven potent patients suffered from penile deviations and were treated via Nesbit's surgical method. Nine long-term impotent patients underwent implantation of flexible hydraulic prothesis. Three potent patients underwent trans-sexual operations. Expression of the sGC and cGK I were examined immunohistochemically using specific antibodies. In all specimens of cavernosal tissue a distinct immunoreactivity was observed in different parts and structures. We found a high expression of sGC and cGK I in smooth muscle cells of vessels and in the fibromuscular stroma. The endothelium of the cavernosal sinus, of the cavernosal arteries, and the cavernosal nerve fibers showed an immunoreactivity against sGC. The distribution analysis of cGK I revealed a predominately vesicular localization in smooth muscle cells. The examination of the endothelium showed no clear immunoreactivity against cGK I. There was no distinct difference in immunoreactivity and cellular distribution between potent and impotent patients. PMID- 11045910 TI - Efficacy of sildenafil in erectile dysfunction after radical prostatectomy. AB - Radical retropubic prostatectomy (RRP) is an important cause of iatrogenic erectile dysfunction (ED). While sildenafil has been widely used since its introduction as a new treatment option for ED, its efficacy in post-RRP patients has not been extensively studied. We retrospectively compared the efficacy of sildenafil in post-RRP and non-surgical patients with ED (NSED) using a subset of questions from the International Index of Erectile Function (IIEF) and correlated results with their specific etiology of ED based on penile blood flow study (PBFS). A brief questionnaire regarding satisfaction with sildenafil was administered to 72 consecutive post-RRP patients (nerve sparing status unknown) and 32 consecutive NSED patients who had previously undergone PBFS with pharmacotesting as part of their evaluation for ED. PBFS diagnoses were arterial insufficiency (AI) for peak systolic velocity (PSV) < 25 cm/sec; venogenic (CVOD) for PSV > or = 35 cm/sec, mixed vascular for PV > 25 but < 35 cm/sec and resistive index (RI) < 0.9; a vascular normal diagnosis (neurogenic impotence) required excellent rigidity sustained for 20 min. Differences in the IIEF subscores for the different groups of patients were assessed. Success with sildenafil was defined as moderate or excellent improvement (3/4 or 4/4) with ability for penetration. No differences were found among the different subgroups of RRP patients with respect to IIEF scores or success rates with sildenafil. NSED patients had both significantly higher post-treatment IIEF scores (3.6/3.4 vs 2.5/2.2; t=4.50, P<0.0001) and success rates (63% vs 31%; t=3.11, P < 0.01) with sildenafil treatment than RRP patients. We found that sildenafil is significantly less effective in impotent RRP patients than in age-matched patients with ED (31% vs 63%). We had postulated that sildenafil would be least effective among RRP patients with excellent sustained rigidity to PGE1, as this subgroup is likely to have neurogenic impotence. We found that sildenafil response rates among subgroups of RRP patients were statistically similar regardless of PBFS diagnosis. IIEF scores for the RRP subgroups were similar but statistically lower than in men with ED and no history of RRP. While individuals with normal vascular responses to PGE1 have an increased likelihood of having neurogenic impotence, in RRP patients, we were unable to demonstrate any difference in efficacy of sildenafil, regardless of the PBFS diagnosis. PMID- 11045911 TI - Treatment of Peyronie's disease with oral colchicine: long-term results and predictive parameters of successful outcome. AB - As recent clinical and animal studies have indicated, colchicine, with its anti fibrotic, anti-mitotic and anti-inflammatory activities, has suppressive effects in the pathogenesis of Peyronie's disease. Oral colchicine treatment was initiated in 60 Peyronie's patients during their acute phase (mean duration of disease: 5.7 +/- 4.3 months). Long-term results, based on changes of subjective and objective criteria, were assessed and predictive factors of successful outcome were investigated. After a mean follow-up of 10.7 +/- 4.7 months, the penile deformity improved in 30%, remained unchanged in 48.3% and deteriorated in 21.7%. Pain resolved in 95%. Best results were obtained in those with no risk factor for vascular disease, presenting during the initial 6 months of disease, degree of curvature <30 degrees, no erectile dysfunction by history and positive response to combined injection and stimulation test. In conclusion since tunica albuginea is affected as a whole in Peyronie's disease, systemic oral agents, such as colchicine, may be preferred in the early phase of the disease. PMID- 11045912 TI - Long-term efficacy and safety of oral Viagra (sildenafil citrate) in men with erectile dysfunction and the effect of randomised treatment withdrawal. AB - The long-term efficacy and safety of oral Viagra (sildenafil citrate), a selective phosphodiesterase 5 inhibitor, and the effect of withdrawing treatment were evaluated in men with erectile dysfunction (ED). In 233 men with ED of psychogenic or mixed organic/psychogenic aetiology, 16 weeks of open-label, flexible-dose sildenafil treatment (10-100 mg) was followed by eight weeks of double-blind, fixed-dose, randomised withdrawal to placebo or continued treatment with sildenafil. Sildenafil was taken as needed (not more than once daily) approximately 1 h prior to sexual activity. The main outcome measures were a global efficacy question, a sexual function questionnaire, an event log of erections, and adverse event recording. In the open-label phase, 200 of 216 patients (93%) reported improved erections with sildenafil; 28 patients (12%) discontinued treatment. In the double-blind phase, the significant improvements in the frequency and duration of erections were maintained in the sildenafil group but returned to pre-treatment values in patients on placebo (P values < 0.0001 versus placebo). The most frequent adverse events in the sildenafil group during the double-blind phase were flushing (7%), headache (6%), and dyspepsia (5%). Of the 192 patients enrolled in the 1-y extension, 90% completed the study; only two patients (1%) were withdrawn due to lack of efficacy. In men with ED of psychogenic or mixed aetiology, oral sildenafil is effective and well-tolerated both at the initiation of therapy and during long-term treatment. For most patients, sildenafil treatment must be continued for improvements in erectile function to be maintained. PMID- 11045913 TI - Pre-penile arteries are dominant in the regulation of penile vascular resistance in the rat. AB - The amount of blood flow into the penis that will produce an erection is dependent on the sum of inflow resistance from the feeder arteries, arterioles and the intra-penile vasculature. In the present study, our objective was to determine quantitatively the contribution to inflow resistance of these different components of the rat penile vasculature. Using methods developed previously, we determined the resistance properties of the isolated perfused whole penis in situ, both in an intact system and after serial transactions of the vessels. These cuts eliminated progressively larger distal segments of the vascular bed. Perfusion pressures were recorded at different flow rates (0.5-3 ml/min/kg body weight) under conditions of maximal dilatation and maximal vasoconstriction induced by methoxamine (MXA, 40 microg/ml). Regardless of the level of vascular tone, the pudendal artery contributes approximately 70% of the total resistance of the penile vasculature. In contrast, the vasculature within the penis (tip, shaft, crus) contributes only about one quarter of the resistance. Penile arterial inflow resistance properties both at maximal vasodilation and maximal alpha1-adrenergic constriction are dominated by the extra-penile vasculature in the rat. The implications of these findings are that alterations in the pudendal artery (eg vasodilation, vasoconstriction, stenosis) would have primary control of arterial inflow and suggest an important role for pharmacological agents which can promote a more generalized vasodilation (eg phosphodiesterase inhibitors) in contrast to selective corpus cavernosal agents. PMID- 11045914 TI - Prostaglandin E1 versus sex therapy in the management of psychogenic erectile dysfunction. AB - The treatment for psychogenic erectile dysfunction has been previously managed by non-medical methods consisting of counseling with a psychiatrist, psychologist or sex therapist. The success rate for treatment with counseling has not been uniformly successful. This paper compares the treatment of psychogenic erectile dysfunction using standard sex therapy and self-injection therapy using low-dose PGE1. Fifty men with psychogenic impotence were divided into two groups: standard sex therapy for twelve weeks or treatment using low-dose (2.5 - 5.0 microg) of PGE1. The results showed that men treated with low-dose PGE1 had a 47% improvement of obtaining an unaided erection compared to 58% improvement rate with sex therapy. 69% of patients in the PGE1 group were satisfied with their treatment compared to 75% receiving sex therapy. The frequency of intercourse reported in patient diaries for the two groups was similar (20.5 per month for PGE1 vs 20.0 per month for sex therapy. The reported duration of erection by patients receiving PGE1 therapy was longer than that reported by those receiving sex therapy (35 min vs 10 min). The comparison of the cost of treatment of the two treatment groups reveals that the sex therapy is approximately 25% more expensive than the PGE1 treatment. This pilot study demonstrates that the efficacy of PGE1 was numerically, though not statistically, less than sex therapy in the treatment of psychogenic impotence. The cost per positive outcome with PGE1 treatment is lower than that of sex therapy treatment making PGE1 more cost effective. PMID- 11045915 TI - Use of transurethral alprostadil (MUSE) (prostaglandin E1) for glans tumescence in a patient with penile prosthesis. AB - The presence of communication between the emissary veins from the corpora cavernosa and the circumflex veins draining the corpus spongiosum makes it possible for the transfer of alprostadil (prostaglandin E1) in MUSE from the spongiosal compartment to the cavernosal compartment of the penis after its absorption through the urethral mucosa. This leads to engorgement and tumescence of the corpus spongiosum as well as the corpora cavernosa. Lack of tumescence of the glans penis and poor penile girth can be a cause for disappointment and frustration in patients following penile prosthetic surgery. MUSE was used successfully in a patient with a Dynaflex penile prosthesis to enhance the tumescence of the glans penis. It will be a useful adjunct for patients in similar circumstances. PMID- 11045916 TI - Genome of a microbial organism. PMID- 11045917 TI - Proteomics, DNA arrays and the analysis of still unknown regulons and unknown proteins of Bacillus subtilis and pathogenic gram-positive bacteria. AB - The complete sequence of the bacterial genomes provides new perspectives for the study of gene expression and gene function. By the combination of the highly sensitive 2-dimensional (2D) protein gel electrophoresis with the identification of the protein spots by microsequencing or mass spectrometry we established a 2D protein index of Bacillus subtilis that currently comprises almost 400 protein entries. A computer-aided evaluation of the 2D gels loaded with radioactively labelled proteins from growing or stressed/starved cells proved to be a powerful tool in the analysis of global regulation of the expression of the entire genome. For the general stress regulon it is demonstrated how the proteomics approach can be used to analyse the regulation, structure and function of still unknown regulons. The application of this approach is illustrated for the sigmaB dependent general stress regulon. For the comprehensive description of proteins/genes belonging to stimulons or regulons it is generally recommended to complement the proteome approach with DNA array techniques in order to identify and allocate still undiscovered members of individual regulons. This approach is also very attractive to uncover the function of still unknown global regulators and regulons and to dissect the entire genome into its basic modules of global regulation. The same strategy can be used to analyse the regulation, structure and function of regulons encoding virulence factors of pathogenic bacteria for a comprehensive understanding of the pathogenicity and for the identification of new antibacterial targets. PMID- 11045918 TI - Rickettsia prowazekii and Bartonella henselae: differences in the intracellular life styles revisited. AB - Within the alpha subdivision of proteobacteria, the arthropod-borne human pathogens Rickettsia prowazekii and Bartonella henselae provide examples of bacteria with obligate and facultative intracellular life styles, respectively. The complete genome sequence of R. prowazekii has been published, whereas the sequencing of the B. henselae genome is in its final stage. Here, we provide a brief overview of a comparative analysis of both genomes based on the delineated metabolic properties. The relative proportion of genes devoted to basic information processes is similar in the two genomes. In contrast, a full set of genes encoding proteins involved in the biosynthesis of amino acids and nucleotides is present in B. henselae, while the majority of these genes is absent from R. prowazekii. This suggests that B. henselae has a better potential for growth in the free-living mode, whereas R. prowazekii is more specialised to growth in an intracellular environment. Functional genomics will provide the potential to further resolve the genetic basis for successful human infections by these important parasites. PMID- 11045919 TI - Comparative genomics of the mycobacteria. AB - The genus mycobacteria includes two important human pathogens Mycobacterium tuberculosis and Mycobacterium lepra. The former is reputed to have the highest annual global mortality of all pathogens. Their slow growth, virulence for humans and particular physiology makes these organisms extremely difficult to work with. However the rapid development of mycobacterial genomics following the completion of the Mycobacterium tuberculosis genome sequence provides the basis for a powerful new approach for the understanding of these organisms. Five further genome sequencing projects of closely related mycobacterial species with differing host range, virulence for humans and physiology are underway. A comparative genomic analysis of these species has the potential to define the genetic basis of these phenotypes which will be invaluable for the development of urgently needed new vaccines and drugs. This minireview summarises the different techniques that have been employed to compare these genomes and gives an overview of the wealth of data that has already been generated by mycobacterial comparative genomics. PMID- 11045920 TI - Genome plasticity in Enterobacteriaceae. AB - The comparative analysis of multiple representatives of the genomes of particular species are leading us away from a view of bacterial genomes as static, monolithic structures towards the view that they are relatively variable, fluid structures. This plasticity is mainly the result of the rearrangement of genes within the genome and the acquisition of novel genes by horizontal transfer systems, e. g. plasmids, bacteriophages, transposons or gene cassettes. These mechanisms often act in concert thus generating a complex genetic structure. Genomic variations are not a phenomenon at the DNA level alone, they influence the phenotype of a bacterium as well and can render a formerly harmless organism into a hazardous pathogen. This review deals not only with the mechanisms of genome rearrangements and the horizontal transfer of genes in Enterobacteriaceae but also points out that mobile genetic elements themselves are subjected to variation. PMID- 11045921 TI - Genome organization and the evolution of the virulence gene locus in Listeria species. AB - The chromosomal region of Listeria monocytogenes harboring the gene cluster prfA plcA-hly-mpl-actA-plcB (virulence gene cluster; vgc) harbors virulence genes critical for the survival of the bacteria following infection. Previous studies have implicated it as an ancestral pathogenicity island, derivatives of which are present in the species L. ivanovii and L. seeligeri, but absent in non-pathogenic species such as L. innocua. We cloned the corresponding region from L. innocua and L. welshimeri and compared its sequences to those from L. monocytogenes, L. ivanovii and L. seeligeri. The analysis allowed exact determination of delineation and size of the vgc and suggests that these genes may have been acquired by bacteriophage transduction. Thus, here we present an alternative view of the evolution of Listeria spp. and suggest that L. monocytogenes may be the primordial species of this genus. PMID- 11045922 TI - Genetic variability within Helicobacter pylori. AB - One of the striking characteristics of the human gastric pathogen Helicobacter pylori is extensive genetic diversity between different strains. During the last years, intensive research using genomic, molecular genetic and population genetic approaches has permitted to quantitate the diversity in H. pylori, to define its characteristics, and to understand the mechanisms that generate diversity in this global pathogen. This review summarizes the recent developments in this rapidly moving field and discusses the significance of genomic heterogeneity and nucleotide sequence diversity in the contexts of H. pylori epidemiology and pathogenesis. PMID- 11045923 TI - High-throughput sequencing in the population analysis of bacterial pathogens of humans. AB - High-throughput nucleotide sequence determination technologies present new opportunities for studies of bacterial pathogens by enabling the accumulation of large volumes of biodiversity information from isolate collections. Population studies, which combine these data with epidemiological, phylogenetic, and evolutionary concepts, provide insights into the behaviour of pathogens that are unavailable from other approaches as they address questions of relevance to pathogenesis from the perspective of the infectious organism rather from that of the host. Hypothesis-driven analyses applied to these data permit the determination of microbial population diversity and structure, the identification of the mechanisms of genetic change in bacterial populations, and the generation of models of pathogen evolution. The nucleotide sequence-based population studies performed to date demonstrate a spectrum of nucleotide sequence diversity, population structure, and evolutionary mechanisms among pathogenic bacteria. The rapid development of nucleotide sequence determination and analysis techniques provides the tools necessary for the prosecution of population studies on an increasing number of bacterial pathogens. PMID- 11045925 TI - The Anthropology of Religion and the Quarrel between Poetry and Philosophy. PMID- 11045924 TI - Target-based drug discovery for the development of novel antiinfectives. AB - In the 20th century and especially during the last 50 years, antiinfectives have been increasingly used to control and prevent infectious diseases. Unfortunately the resistance of microorganisms to these pharmaceuticals has increased as well. At the same time the discovery process for novel antiinfectives, the so-called "conventional" screening approach, involves testing natural products or derivatives of known compounds in in vitro cultures. By now it is obvious that this screening approach did not meet the expectations to generate a sufficient number of novel drug candidates. Consequently, studies for selective antiinfectives with new modes of action, which are able to break resistance, are highly desirable for human and animal health. The enormous advance in sequencing technologies--leading to a constantly growing number of known microbial genomes- together with the rapid development of computer power and bioinformatic software tools, now makes it possible to identify genes and gene products that are essential to the pathogenic organisms and are therefore considered to be novel targets for the development of new antiinfectives. When these potential targets have been validated by sophisticated laboratory methods, large diverse compound libraries can be tested in in vitro assays using high-throughput screening. This approach will most likely generate an increasing number of novel lead structures that will be specifically optimized by modern combinatorial chemistry and subsequently lead to new antiinfective candidates strengthening the armoury of weapons available to fight infectious diseases in humans and animals. PMID- 11045926 TI - The Language of Classic Maya Inscriptions1. AB - Recent decipherments of Classic Maya hieroglyphs (ca. a.d. 250 to 850) reveal phonological and morphological patterns that, through epigraphic and historical analyses, isolate a single, coherent prestige language with unique and widespread features in script. We term this language "Classic Ch'olti'an" and present the evidence for its explicable historical configuration and ancestral affiliation with Eastern Ch'olan languages (Ch'olti' and its still-viable descendant, Ch'orti'). We conclude by exploring the possibility that Ch'olti'an was a prestige language that was shared by elites, literati, and priests and had a profound effect on personal and group status in ancient Maya kingdoms. PMID- 11045927 TI - Genes, Tribes, and African History. AB - Over the past 40 years, traditional perspectives on the constitution of human groups have been subjected to stringent critique within anthropology. This began with the dismantling of accepted "race" divisions after World War II and continued with analyses of the meaning and reality of African "tribal" distinctions from the 1960s until the present. Archaeologists, ethnographers, linguists, and historians of Africa now work within a research milieu where social interactions, cultural exchange, and the dynamic nature of group identifications are accepted as a normal part of the human experience. At the same time, new techniques have been developed for the examination of human history, techniques based upon an expanding repertoire of tools for the analysis of genetic variability in human populations. Perhaps the most striking result of this research has been Cavalli-Sforza, Menozzi, and Piazza's The History and Geography of Human Genes. Rather less attention has been paid, however, to the conceptual relationships between the human groups defined through such analyses, in Africa and elsewhere, and those defined through other kinds of research. This paper is a preliminary examination of the fit between genetic, archaeological, and ethnographic data on the African past. PMID- 11045929 TI - Essential Differences: National Homogeneity and Cultural Representation in Four Recent Works on Greek Macedonia. PMID- 11045928 TI - The Evolution of Human Homosexual Behavior. AB - Homosexuality presents a paradox for evolutionists who explore the adaptedness of human behavior. If adaptedness is measured by reproductive success and if homosexual behavior is nonreproductive, how has it come about? Three adaptationist hypotheses are reviewed here and compared with the anthropological literature. There is little evidence that lineages gain reproductive advantage through offspring care provided by homosexual members. Therefore, there is little support for the hypothesis that homosexuality evolved by kin selection. Parents at times control children's reproductive decisions and at times encourage children in homosexual behavior. There is therefore more support for the hypothesis of parental manipulation. Support is strongest, however, for the hypothesis that homosexual behavior comes from individual selection for reciprocal altruism. Same-sex alliances have reproductive advantages, and sexual behavior at times maintains these alliances. Nonhuman primates, including the apes, use homosexual behavior in same-sex alliances, and such alliances appear to have been key in the expanded distribution of human ancestors during the Pleistocene. Homosexual emotion and behavior are, in part, emergent qualities of the human propensity for same-sex affiliation. Adaptationist explanations do not fully explain sexual behavior in humans, however; social and historical factors also play strong roles. PMID- 11045930 TI - The Anthropology of Religion and the Quarrel between Poetry and Philosophy. PMID- 11045931 TI - The Language of Classic Maya Inscriptions1. AB - Recent decipherments of Classic Maya hieroglyphs (ca. a.d. 250 to 850) reveal phonological and morphological patterns that, through epigraphic and historical analyses, isolate a single, coherent prestige language with unique and widespread features in script. We term this language "Classic Ch'olti'an" and present the evidence for its explicable historical configuration and ancestral affiliation with Eastern Ch'olan languages (Ch'olti' and its still-viable descendant, Ch'orti'). We conclude by exploring the possibility that Ch'olti'an was a prestige language that was shared by elites, literati, and priests and had a profound effect on personal and group status in ancient Maya kingdoms. PMID- 11045932 TI - Genes, Tribes, and African History. AB - Over the past 40 years, traditional perspectives on the constitution of human groups have been subjected to stringent critique within anthropology. This began with the dismantling of accepted "race" divisions after World War II and continued with analyses of the meaning and reality of African "tribal" distinctions from the 1960s until the present. Archaeologists, ethnographers, linguists, and historians of Africa now work within a research milieu where social interactions, cultural exchange, and the dynamic nature of group identifications are accepted as a normal part of the human experience. At the same time, new techniques have been developed for the examination of human history, techniques based upon an expanding repertoire of tools for the analysis of genetic variability in human populations. Perhaps the most striking result of this research has been Cavalli-Sforza, Menozzi, and Piazza's The History and Geography of Human Genes. Rather less attention has been paid, however, to the conceptual relationships between the human groups defined through such analyses, in Africa and elsewhere, and those defined through other kinds of research. This paper is a preliminary examination of the fit between genetic, archaeological, and ethnographic data on the African past. PMID- 11045933 TI - The Evolution of Human Homosexual Behavior. AB - Homosexuality presents a paradox for evolutionists who explore the adaptedness of human behavior. If adaptedness is measured by reproductive success and if homosexual behavior is nonreproductive, how has it come about? Three adaptationist hypotheses are reviewed here and compared with the anthropological literature. There is little evidence that lineages gain reproductive advantage through offspring care provided by homosexual members. Therefore, there is little support for the hypothesis that homosexuality evolved by kin selection. Parents at times control children's reproductive decisions and at times encourage children in homosexual behavior. There is therefore more support for the hypothesis of parental manipulation. Support is strongest, however, for the hypothesis that homosexual behavior comes from individual selection for reciprocal altruism. Same-sex alliances have reproductive advantages, and sexual behavior at times maintains these alliances. Nonhuman primates, including the apes, use homosexual behavior in same-sex alliances, and such alliances appear to have been key in the expanded distribution of human ancestors during the Pleistocene. Homosexual emotion and behavior are, in part, emergent qualities of the human propensity for same-sex affiliation. Adaptationist explanations do not fully explain sexual behavior in humans, however; social and historical factors also play strong roles. PMID- 11045934 TI - Essential Differences: National Homogeneity and Cultural Representation in Four Recent Works on Greek Macedonia. PMID- 11045935 TI - Vascular endothelial growth factor is modulated in vascular muscle cells by estradiol, tamoxifen, and hypoxia. AB - Vascular endothelial growth factor (VEGF) promotes neovascularization, microvascular permeability, and endothelial proliferation. We described previously VEGF mRNA and protein induction by estradiol (E2) in human endometrial fibroblasts. We report here E2 induction of VEGF expression in human venous muscle cells [smooth muscle cells (SMC) from human saphenous veins; HSVSMC] expressing both ER-alpha and ER-beta estrogen receptors. E2 at 10(-9) to 10(-8) M increases VEGF mRNA in HSVSMC in a time-dependent manner (3-fold at 24 h), as analyzed by semiquantitative RT-PCR. This level of induction is comparable with E2 endometrial induction of VEGF mRNA. Tamoxifen and hypoxia also increase HSVSMC VEGF mRNA expression over control values. Immunocytochemistry of saphenous veins and isolated SMC confirms translation of VEGF mRNA into protein. Immunoblot analysis of HSVSMC-conditioned medium detects three bands of 18, 23, and 28 kDa, corresponding to VEGF isoforms of 121, 165, and 189 amino acids. Radioreceptor assay of the conditioned medium produced by E2-stimulated HSVSMC reveals an increased VEGF secretion. Our data indicate that VEGF is E2, tamoxifen, and hypoxia inducible in cultured HSVSMC and E2 inducible in aortic SMC, suggesting E2 modulation of VEGF effects in angiogenesis, vascular permeability, and integrity. PMID- 11045936 TI - High spatial resolution measurements of organ blood flow in small laboratory animals. AB - With the use of a newly developed Imaging Cryomicrotome to determine the spatial location of fluorescent microspheres in organs, we validate and report our processing algorithms for measuring regional blood flow in small laboratory animals. Microspheres (15-microm diameter) of four different fluorescent colors and one radioactive label were simultaneously injected into the left ventricle of a pig. The heart and kidneys were dissected, and the numbers of fluorescent and radioactive microspheres were determined in 10 randomly selected pieces. All microsphere counts fell well within the 95% expected confidence limits as determined from the radioactive counts. Fluorescent microspheres (15-microm diameter) of four different colors were also injected into the tail vein of a rat and the left ventricle of a rabbit. After correction for Poisson noise, correlation coefficients between the colors were 0.99 +/- 0.02 (means +/- SD) for the rabbit heart and 0.99 +/- 0.02 for the rat lung. Mathematical dissection algorithms, statistics to analyze the spatial data, and methods to visualize blood flow distributions in small animal organs are presented. PMID- 11045937 TI - Endotoxin infusion in rats induces apoptotic and survival pathways in hearts. AB - Inflammatory mediators of sepsis induce apoptosis in many cell lines. We tested the hypothesis that lipopolysaccharide (LPS) injection in vivo results in induction of early apoptotic and survival pathways as well as evidence of late stage apoptosis in the heart. Hearts were collected from control rats and at 6, 12, and 24 h after LPS injection (4 mg/kg). Activation of an apoptotic pathway was identified by a 1,000-fold increase in caspase-3 activity at 24 h (P < 0.05). Confirmation of these results occurred when terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) staining identified myocardial cells undergoing DNA fragmentation with significant levels at 24 h post-LPS injection. LPS also caused early proapoptotic mRNA (Bax) to increase (16% at 24 h, P < 0.05), whereas the Bax protein initially decreased (35% at 6 h, P < 0.05) and then returned to baseline values by 24 h. Six hours after LPS injection, Bcl-2 (early prosurvival) mRNA levels increased, whereas its protein levels decreased (70%, P < 0.05) and then returned to baseline levels by 24 h. Mitochondrial cytochrome c levels decreased, suggestive of mitochondrial involvement. Thus involvement of proapoptotic and prosurvival pathways in the heart occurs during a septic inflammatory response. PMID- 11045938 TI - Genetic mapping of quantitative trait loci influencing left ventricular mass in rats. AB - High blood pressure is the leading cause of left ventricular hypertrophy (LVH); however, not all hypertensive patients develop LVH. Genetic factors are important in the development of LVH. With the use of F2 male rats from spontaneously hypertensive rats and Lewis rats, we performed a study to identify the quantitative trait loci (QTL) that influence left ventricular mass (LVM). Mean arterial pressure (MAP) was measured by the direct intra-arterial method in conscious animals, and LVM was determined at 24 wk of age. QTL analysis was done using 160 microsatellite markers for a genome-wide scan. Two loci that influenced body weight-adjusted LVM with logarithm of the odds scores >3.4 were found. One locus on chromosome 17 influenced LVM independently of MAP. Another locus on chromosome 7 influenced LVM and MAP. These findings indicate not only the existence of a gene on chromosome 7 that influences LVM in a manner dependent on blood pressure but also the existence of a gene on chromosome 17 that influences LVM independently of blood pressure. PMID- 11045939 TI - SOD-1 expression in pig coronary arterioles is increased by exercise training. AB - Coronary arterioles of exercise-trained (EX) pigs have enhanced nitric oxide (NO.)-dependent dilation. Evidence suggests that the biological half-life of NO. depends in part on the management of the superoxide anion. The purpose of this study was to test the hypothesis that expression of cytosolic copper/zinc dependent superoxide dismutase (SOD)-1 is increased in coronary arterioles as a result of exercise training. Male Yucatan pigs either remained sedentary (SED, n = 4) or were EX (n = 4) on a motorized treadmill for 16-20 wk. Individual coronary arterioles ( approximately 100-microm unpressurized internal diameter) were dissected and frozen. Coronary arteriole SOD-1 protein (via immunoblots) increased as a result of exercise training (2.16 +/- 0.35 times SED levels) as did SOD-1 enzyme activity (measured via inhibition of pyrogallol autooxidation; approximately 75% increase vs. SED). In addition, SOD-1 mRNA levels (measured via RT-PCR) were higher in EX arterioles (1.68 +/- 0.16 times the SED levels). There were no effects of exercise training on the levels of SOD-2 (mitochondrial), catalase, or p67(phox) proteins. Thus chronic aerobic exercise training selectively increases the levels of SOD-1 mRNA, protein, and enzymatic activity in porcine coronary arterioles. Increased SOD-1 could contribute to the enhanced NO.-dependent dilation previously observed in EX porcine coronary arterioles by improving management of superoxide in the vascular cell environment, thus prolonging the biological half-life of NO. PMID- 11045940 TI - Effect of time and vascular pressure on permeability and cyclic nucleotides in ischemic lungs. AB - We previously found that increased intravascular pressure decreased ischemic lung injury by a nitric oxide (NO)-dependent mechanism (Becker PM, Buchanan W, and Sylvester JT. J Appl Physiol 84: 803-808, 1998). To determine the role of cyclic nucleotides in this response, we measured the reflection coefficient for albumin (sigma(alb)), fluid flux (), cGMP, and cAMP in ferret lungs subjected to either 45 min ("short"; n = 7) or 180 min ("long") of ventilated ischemia. Long ischemic lungs had "low" (1-2 mmHg, n = 8) or "high" (7-8 mmHg, n = 6) vascular pressure. Other long low lungs were treated with the NO donor (Z)-1-[N-(3-ammoniopropyl)-N (n-propyl)amino]diazen-1-ium -1, 2-diolate (PAPA-NONOate; 5 x 10(-4) M, n = 6) or 8-bromo-cGMP (5 x 10(-4) M, n = 6). Compared with short ischemia, long low ischemia decreased sigma(alb) (0.23 +/- 0.04 vs. 0.73 +/- 0.08; P < 0.05) and increased (1.93 +/- 0.26 vs. 0.58 +/- 0.22 ml. min(-1). 100 g(-1); P < 0.05). High pressure prevented these changes. Lung cGMP decreased by 66% in long compared with short ischemia. Lung cAMP did not change. PAPA-NONOate and 8-bromo cGMP increased lung cGMP, but only 8-bromo-cGMP decreased permeability. These results suggest that ischemic vascular injury was, in part, mediated by a decrease in cGMP. Increased vascular pressure prevented injury by a cGMP independent mechanism that could not be mimicked by administration of exogenous NO. PMID- 11045941 TI - Increased medial smooth muscle cell length is responsible for vascular hypertrophy in young hypertensive rats. AB - Large mesenteric arteries from 3- to 4-wk-old spontaneously hypertensive rats (SHR) showed medial hypertrophy and an increased contractile response to various agonists before significant blood pressure increase. Here we determined the cellular nature of this vascular hypertrophy. Large mesenteric arteries from SHR and Wistar-Kyoto (WKY) rats were fixed at maximal relaxation either with an in situ perfusion fixation or an in vitro fixation method. With the use of morphometric protocols and confocal microscopy, the volume of the medial wall and lumen, numerical density of smooth muscle cell nuclei in the medial layer, and smooth muscle cell and nuclear length were measured. Both methods of fixation yielded similar results, showing significant medial volume expansion in SHR than WKY without lumen change. Numerical density of medial smooth muscle cells was significantly less in SHR than WKY, and their total number per 100 microm length were similar between the strains. Average smooth muscle nuclear and cell length from SHR was significantly longer than that of WKY. Regression analysis showed that the increase in smooth muscle cell length explained 80% of the medial volume increase. We concluded that increased smooth muscle cell length in prehypertensive SHR is responsible for increased medial volume in the mesenteric arteries. PMID- 11045942 TI - Fast pacing facilitates discontinuous action potential propagation between rabbit atrial cells. AB - We examined the critical coupling conductance (G(C)) for propagation at different pacing cycle lengths (CLs) (1,000 and 400 ms). As G(C) was progressively reduced, propagation failed at a CL of 1,000 ms, whereas propagation succeeded at a CL of 400 ms over a range of G(C) values before failing at a CL of 400 ms at a lower G(C), showing facilitation of propagation at the shorter CL. Critical G(C) was (means +/- SE) 0.8 +/- 0.1 nS for a CL of 400 ms and 1.3 +/- 0.1 nS for a CL of 1,000 ms (a 63% increase, P < 0.002, n = 9 cell pairs). In 14 uncoupled cells, action potential duration at 30% repolarization (APD(30)) increased from 19.9 +/- 2.5 to 41.8 +/- 2.6 ms (P < 0.001) as CL decreased from 1,000 to 400 ms. In five cell pairs, critical G(C) with 4-aminopyridine (4-AP) was reduced to 0.4 +/- 0.1 nS at a CL of 1,000 ms (P < 0.05 compared with control solution), and critical G(C) in 4-AP was unchanged by decreasing CL to 400 ms. It is possible that the "remodeling" of atrial cells due to atrial fibrillation or tachycardia, which has been shown to produce a decrease in the transient outward current, may result in an enhanced ability to propagate, possibly facilitating further development of fibrillation under conditions of decreased cellular coupling. PMID- 11045943 TI - Regulation of a voltage-sensitive release mechanism by Ca(2+)-calmodulin dependent kinase in cardiac myocytes. AB - A role for Ca(2+)-calmodulin-dependent kinase (CamK) in regulation of the voltage sensitive release mechanism (VSRM) was investigated in guinea pig ventricular myocytes. Voltage clamp was used to separate the VSRM from Ca(2+)-induced Ca(2+) release (CICR). VSRM contractions and Ca(2+) transients were absent in cells dialyzed with standard pipette solution but present when 2-5 microM calmodulin was included. Effects of calmodulin were blocked by KN-62 (CamK inhibitor), but not H-89, a protein kinase A (PKA) inhibitor. Ca(2+) current and caffeine contractures were not affected by calmodulin. Transient-voltage relations were bell-shaped without calmodulin, but they were sigmoidal and typical of the VSRM with calmodulin. Contractions with calmodulin exhibited inactivation typical of the VSRM. These contractions were inhibited by rapid application of 200 microM of tetracaine, but not 100 microM of Cd(2+), whereas CICR was inhibited by Cd(2+) but not tetracaine. In undialyzed myocytes (high-resistance microelectrodes), KN 62 or H-89 each reduced amplitudes of VSRM contractions by approximately 50%, but together they decreased VSRM contractions by 93%. Thus VSRM is facilitated by CamK or PKA, and both pathways regulate the VSRM in undialyzed cells. PMID- 11045944 TI - Inhibition of adenosine kinase induces expression of VEGF mRNA and protein in myocardial myoblasts. AB - We tested whether increased endogenous adenosine produced by the adenosine kinase inhibitor GP-515 (Metabasis Therapeutics) can induce vascular endothelial growth factor (VEGF) expression in cultured rat myocardial myoblasts (RMMs). RMMs were cultured for 18 h in the absence (control) and presence of GP-515, adenosine (Ado), adenosine deaminase (ADA), or GP-515 + ADA. GP-515 (0.2-200 microM) caused a dose-related increase in VEGF protein expression (1.99-2.84 ng/mg total cell protein); control VEGF was 1.84 +/- 0.05 ng/mg. GP-515 at 2 and 20 microM also increased VEGF mRNA by 1.67- and 1. 82-fold, respectively. ADA (10 U/ml) decreased baseline VEGF protein levels by 60% and completely blocked GP-515 induction of VEGF. Ado (20 microM) and GP-515 (20 microM) caused a 59 and 39% increase in VEGF protein expression and a 98 and 33% increase in human umbilical vein endothelial cell proliferation, respectively, after 24 h of exposure. GP-515 (20 microM) had no effect on VEGF protein expression during severe hypoxia (1% O(2)) but increased VEGF by an additional 27% during mild hypoxia (10% O(2)). These results indicate that raising endogenous levels of Ado through inhibition of adenosine kinase can increase the expression of VEGF and stimulate endothelial cell proliferation during normoxic and hypoxic conditions. PMID- 11045945 TI - A metabolic role for mitochondria in palmitate-induced cardiac myocyte apoptosis. AB - After cardiac ischemia, long-chain fatty acids, such as palmitate, increase in plasma and heart. Palmitate has previously been shown to cause apoptosis in cardiac myocytes. Cultured neonatal rat cardiac myocytes were studied to assess mitochondrial alterations during apoptosis. Phosphatidylserine translocation and caspase 3-like activity confirmed the apoptotic action of palmitate. Cytosolic cytochrome c was detected at 8 h and plateaued at 12 h. The mitochondrial membrane potential (DeltaPsi) in tetramethylrhodamine ethyl ester-loaded cardiac myocytes decreased significantly in individual mitochondria by 8 h. This loss was heterogeneous, with a few energized mitochondria per myocyte remaining at 24 h. Total ATP levels remained high at 16 h. The DeltaPsi loss was delayed by cyclosporin A, a mitochondrial permeability transition inhibitor. Mitochondrial swelling accompanied changes in DeltaPsi. Carnitine palmitoyltransferase I activity fell at 16 h; this decline was accompanied by ceramide increases that paralleled decreased complex III activity. We conclude that carnitine palmitoyltransferase I inhibition, ceramide accumulation, and complex III inhibition are downstream events in cardiac apoptosis mediated by palmitate and occur independent of events leading to caspase 3-like activation. PMID- 11045946 TI - Phenotypical features of long Q-T syndrome in transgenic mice expressing human Na K-ATPase alpha(3)-isoform in hearts. AB - To understand why the adult human heart expresses three isoforms of the sodium pump, we generated transgenic mice (TGM) with 2.3- to 5. 5-fold overexpression of the human alpha(3)-isoform of Na-K-ATPase in the heart. Hearts from the TGM had increased maximal Na-K-ATPase activity and ouabain affinity compared with control hearts, even though the density of Na-K-ATPase pump sites (of all isoforms) was similar to that of control mice. In perfused hearts, contractility both at baseline and in the presence of ouabain tended to be greater in TGM than in controls. Surface electrocardiograms in anesthetized TGM had a steeper dependence of Q-T on sinus cycle length, and Q-T intervals measured during atrial pacing were significantly longer in TGM. Q-T dispersion during sinus rhythm also tended to be longer in TGM. Thus TGM overexpressing human alpha(3)-isoform have several of the phenotypical features of human long Q-T syndrome, despite the absence of previously described mutations in Na(+) or K(+) channels. PMID- 11045947 TI - Inhibition of NHE protects reoxygenated cardiomyocytes independently of anoxic Ca(2+) overload and acidosis. AB - We investigated the question of whether inhibition of the Na(+)/H(+) exchanger (NHE) during ischemia is protective due to reduction of cytosolic Ca(2+) accumulation or enhanced acidosis in cardiomyocytes. Additionally, the role of the Na(+)-HCO(3)(-) symporter (NBS) was investigated. Adult rat cardiomyocytes were exposed to simulated ischemia and reoxygenation. Cytosolic pH [2', 7'-bis(2 carboxyethyl)-5(6)-carboxyfluorescein (BCECF)], Ca(2+) (fura 2), Na(+) [sodium binding benzolfuran isophthatlate (SBFI)], and cell length were measured. NHE was inhibited with 3 micromol/l HOE 642 or 1 micromol/l 5-(N-ethyl-N-isopropyl) amiloride (EIPA), and NBS was inhibited with HEPES buffer. During anoxia in bicarbonate buffer, cells developed acidosis and intracellular Na and Ca (Na(i) and Ca(i), respectively) overload. During reoxygenation cells underwent hypercontracture (44.0 +/- 4.1% of the preanoxic length). During anoxia in bicarbonate buffer, inhibition of NHE had no effect on changes in intracellular pH (pH(i)), Na(i), and Ca(i), but it significantly reduced the reoxygenation induced hypercontracture (HOE: 61.0 +/- 1.4%, EIPA: 68.2 +/- 1.8%). The sole inhibition of NBS during anoxia was not protective. We conclude that inhibition of NHE during anoxia protects cardiomyocytes against reoxygenation injury independently of cytosolic acidification and Ca(i) overload. PMID- 11045948 TI - Conservation of phosphorylation state of cardiac phosphofructokinase during in vitro hypothermic hypoxia. AB - We investigated the metabolic effects of buffering agents alpha-amino-4-imidazole propionic acid (Histidine), N, N-bis(2-hydroxyethyl)glycine (bicine), N, N-bis(2 hydroxyethyl)-2-aminoethanesulfonic acid (BES) on anaerobic energy production (via glycolysis) and conservation of key regulatory enzyme activity, and phosphofructokinase (PFK) throughout prolonged hypothermic hypoxia in porcine hearts. Hearts from 35 to 40 kg pigs were flushed with one of the following five solutions: St. Thomas' Hospital solution (STHS); modified University of Wisconsin (UW) solution; and three solutions containing modified UW plus 90 mM of histidine, bicine, or BES. The hearts were then stored at 4 degrees C for 10 h. After 10 h of hypothermic hypoxia, lactate values were 6.7-12.9 micromol/g higher than control; this reflected an increase in anaerobic end product of 35-67%. The consequences of enhanced anaerobic metabolism were higher ATP, total adenylate, Energy Charge, and ATP/ADP ratios in most of the buffered groups after 4-10 h cold storage; effectiveness of the buffers employed correlated with buffering capacity (BES proved to be the most effective). PFK remained activated throughout most of the 10-h period in hearts stored with buffers and did not undergo the rapid inactivation experienced by hearts stored in STHS. Conservation of PFK integrity with buffering agents was not related to a pH-mediated event; changes in kinetic parameters suggested that this protection was due to an irreversible posttranslational modification, specifically a dephosphorylation event. PMID- 11045949 TI - Autocrine thrombospondin partially mediates TGF-beta1- induced proliferation of vascular smooth muscle cells. AB - Transforming growth factor (TGF)-beta1 has been implicated in vascular healing responses after mechanical injury. Using cultured rat aortic smooth muscle cells (RASMC), we examined the hypothesis that production and secretion of thrombospondin (TSP) contributes to TGF-beta1-induced proliferation. We found that TGF-beta1 enhanced production and secretion of TSP, with peak levels of secreted TSP observed 24 h after treatment. RASMC treated with TGF-beta1 secreted a mitogenic activity that was transferable in conditioned media and partially inhibited by C6.7, a monoclonal anti-TSP antibody. Exogenous TSP stimulated a proliferative response, with maximal [(3)H]thymidine incorporation occurring 24 h earlier than maximal [(3)H]thymidine incorporation in response to TGF-beta1 treatment. Pretreatment with C6.7 or polyclonal anti-TSP neutralizing antibodies inhibited TGF-beta1-induced proliferation of RASMC. Proliferative responses to TGF-beta1 were also inhibited by pretreatment with an anti-beta(3) integrin monoclonal blocking antibody (F11), RGD peptides, and the anti-alpha(v)beta(3) disintegrin echistatin. Treatment with TSP and TGF-beta1 increased c-Jun NH(2) terminal kinase (JNK)1 activity, with peak effects observed at 15 min and 4 h, respectively. Treatment with C6.7 or F11 inhibited TGF-beta-induced activation of JNK1. In summary, these studies support the hypothesis that TGF-beta-induced JNK1 activation and proliferation of RASMC require secretion of TSP and ligation of alpha(v)beta(3)-integrins. PMID- 11045950 TI - Intrinsic A(1) adenosine receptor activation during ischemia or reperfusion improves recovery in mouse hearts. AB - We assessed the role of A(1) adenosine receptor (A(1)AR) activation by endogenous adenosine in the modulation of ischemic contracture and postischemic recovery in Langendorff-perfused mouse hearts subjected to 20 min of total ischemia and 30 min of reperfusion. In control hearts, the rate-pressure product (RPP) and first derivative of pressure development over time (+dP/dt) recovered to 57 +/- 3 and 58 +/- 3% of preischemia, respectively. Diastolic pressure remained elevated at 20 +/- 2 mmHg (compared with 3 +/- 1 mmHg preischemia). Interstitial adenosine, assessed by microdialysis, rose from approximately 0.3 to 1.9 microM during ischemia compared with approximately 15 microM in rat heart. Nonetheless, these levels will near maximally activate A(1)ARs on the basis of effects of exogenous adenosine and 2-chloroadenosine. Neither A(1)AR blockade with 200 nM 8 cyclopentyl-1,3-dipropylxanthine (DPCPX) during the ischemic period alone nor A(1)AR activation with 50 nM N(6)-cyclopentyladenosine altered rapidity or extent of ischemic contracture. However, ischemic DPCPX treatment significantly depressed postischemic recovery of RPP and +dP/dt (44 +/- 3 and 40 +/- 4% of preischemia, respectively). DPCPX treatment during the reperfusion period alone also reduced recovery of RPP and +dP/dt (to 44 +/- 2 and 47 +/- 2% of preischemia, respectively). These data indicate that 1) interstitial adenosine is lower in mouse versus rat myocardium during ischemia, 2) A(1)AR activation by endogenous adenosine or exogenous agonists does not modify ischemic contracture in murine myocardium, 3) A(1)AR activation by endogenous adenosine during ischemia attenuates postischemic stunning, and 4) A(1)AR activation by endogenous adenosine during the reperfusion period also improves postischemic contractile recovery. PMID- 11045951 TI - Oxidative stress impairs cardiac chemoreflexes in diabetic rats. AB - We investigated the effects of diabetes mellitus and antioxidant treatment on the sensory and reflex function of cardiac chemosensory nerves in rats. Diabetes was induced by streptozotocin (STZ; 85 mg/kg ip). Subgroups of sham- and STZ-treated rats were chronically treated with an antioxidant, vitamin E (60 mg/kg per os daily, started 2 days before STZ). Animals were studied 6-8 wk after STZ injection. We measured renal sympathetic nerve activity (RSNA), mean arterial blood pressure (MABP), and cardiac vagal and sympathetic afferent activities in response to stimulation of chemosensitive sensory nerves in the heart by epicardial application of capsaicin (Caps) and bradykinin (BK). In cardiac sympathetic-denervated rats, Caps and BK (1-10.0 microg) evoked a vagal afferent mediated reflex depression of RSNA and MABP, which was significantly blunted in STZ-treated rats (P < 0.05). In vagal-denervated rats, Caps and BK (1-10.0 microg) evoked a sympathetic afferent-mediated reflex elevation of RSNA and MABP, which also was significantly blunted in STZ-treated rats (P < 0.05). Chronic vitamin E treatment effectively prevented these cardiac chemoreflex defects in STZ-treated rats without altering resting blood glucose or hemodynamics. STZ treated rats with insulin replacement did not exhibit impaired cardiac chemoreflexes. In afferent studies, Caps and BK (0.1 g-10.0 microg) increased cardiac vagal and sympathetic afferent nerve activity in a dose-dependent manner in sham-treated rats. These responses were significantly blunted in STZ-treated rats. Vitamin E prevented the impairment of afferent discharge to chemical stimulation in STZ rats. The following were concluded: STZ-induced, insulin dependent diabetes in rats extensively impairs the sensory and reflex properties of cardiac chemosensitive nerve endings, and these disturbances can be prevented by chronic treatment with vitamin E. These results suggest that oxidative stress plays an important role in the neuropathy of this autonomic reflex in diabetes. PMID- 11045952 TI - NOC/oFQ contributes to age-dependent impairment of NMDA-induced cerebrovasodilation after brain injury. AB - This study characterized the effects of fluid percussion brain injury (FPI) on N methyl-D-aspartate (NMDA)-induced vasodilation and determined the role of nociceptin/orphanin FQ (NOC/oFQ) in such changes as a function of age and time postinsult. FPI elevated cerebrospinal fluid (CSF) NOC/oFQ from 70 +/- 3 to 444 +/- 56 pg/ml ( approximately 10(-10) M) within 1 h and to 1,931 +/- 112 pg/ml within 8 h, whereas values returned to control levels within 168 h in the newborn pig. In contrast, FPI elevated CSF NOC/oFQ from 77 +/- 4 to 202 +/- 16 pg/ml within 1 h and values returned to control levels within 8 h in the juvenile pig. Topical NOC/oFQ (10(-10) M) had no effect on pial artery diameter but attenuated NMDA (10(-8), 10(-6) M)-induced dilation (9 +/- 1 and 16 +/- 1 vs. 5 +/- 1 and 10 +/- 1%) in both age groups. In the newborn, NMDA-induced pial artery dilation was reversed to vasoconstriction within 1 h post-FPI and responses remained impaired for 72 h, but such vasoconstriction was attenuated by pretreatment with [F/G]NOC/oFQ(1-13)-NH(2) (10(-6) M, 1 mg/kg iv), an NOC/oFQ antagonist (9 +/- 1 and 16 +/- 1 vs. -7 +/- 1 and -12 +/- 1 vs -2 +/- 1 and -3 +/- 1% for control, FPI, and FPI pretreated with the NOC/oFQ antagonist). In contrast, in the juvenile, NMDA-induced vasodilation was only attenuated within 1 h post-FPI and returned to control within 8 h. Such dilation was also partially restored by the NOC/oFQ antagonist. These data indicate that NOC/oFQ contributes to impaired NMDA pial artery dilation after FPI. These data suggest that the greater NOC/oFQ release in the newborn versus the juvenile may contribute to age-related differences in FPI effects on excitatory amino acid-induced pial dilation. PMID- 11045953 TI - Leukocyte and endothelial cell adhesion molecules in a chronic murine model of myocardial reperfusion injury. AB - Expression of endothelial and leukocyte cell adhesion molecules is a principal determinant of polymorphonuclear neutrophil (PMN) recruitment during inflammation. It has been demonstrated that pharmacological inhibition of these molecules can attenuate PMN influx and subsequent tissue injury. We determined the temporal expression of alpha-granule membrane protein-40 (P-selectin), endothelial leukocyte adhesion molecule 1 (E-selectin), and intercellular cell adhesion molecule 1 (ICAM-1) after coronary artery occlusion and up to 3 days of reperfusion. The expression of all of these cell adhesion molecules peaked around 24 h of reperfusion. We determined the extent to which these molecules contribute to PMN infiltration by utilizing mice deficient (-/-) in P-selectin, E-selectin, ICAM-1, and CD18. Each group underwent 30 min of in vivo, regional, left anterior descending (LAD) coronary artery ischemia and 24 h of reperfusion. PMN accumulation in the ischemic-reperfused (I/R) zone was assessed using histological techniques. Deficiencies of P-selectin, E-selectin, ICAM-1, or CD18 resulted in significant (P < 0.05) attenuation of PMN infiltration into the I/R myocardium (MI/R). In addition, P-selectin, E-selectin, ICAM-1, and CD18 -/- mice exhibited significantly (P < 0.05) smaller areas of necrosis after MI/R compared with wild-type mice. These data demonstrate that MI/R induces coronary vascular expression of P-selectin, E-selectin, and ICAM-1 in mice. Furthermore, genetic deficiency of P-selectin, E-selectin, ICAM-1, or CD18 attenuates PMN sequestration and myocardial injury after in vivo MI/R. We conclude that P selectin, E-selectin, ICAM-1, and CD18 are involved in the pathogenesis of MI/R injury in mice. PMID- 11045954 TI - Oxygen dependency and precision of cytochrome oxidase signal from full spectral NIRS of the piglet brain. AB - Oxidation changes of the copper A (Cu(A)) center of cytochrome oxidase in the brain were measured during brief anoxic swings at both normocapnia and hypercapnia (arterial PCO(2) approximately 55 mmHg). Hypercapnia increased total hemoglobin from 37.5 +/- 9.1 to 50.8 +/- 12.9 micromol/l (means +/- SD; n = 7), increased mean cerebral saturation (Smc(O(2))) from 65 +/- 4 to 77 +/- 3%, and oxidized Cu(A) by 0.43 +/- 0.23 micromol/l. During the onset of anoxia, there were no significant changes in the Cu(A) oxidation state until Smc(O(2)) had fallen to 43 +/- 5 and 21 +/- 6% at normocapnia and hypercapnia, respectively, and the maximum reduction during anoxia was not significantly different at hypercapnia (1.49 +/- 0.40 micromol/l) compared with normocapnia (1.53 +/- 0.44 micromol/l). Residuals of the least squares fitting algorithm used to convert near-infrared spectra to concentrations are presented and shown to be small compared with the component of attenuation attributed to the Cu(A) signal. From these observations, we conclude that there is minimal interference between the hemoglobin and Cu(A) signals in this model, the Cu(A) oxidation state is independent of cerebral oxygenation at normoxia, and the oxidation after hypercapnia is not the result of increased cerebral oxygenation. PMID- 11045955 TI - Mechanism of adenosine-induced vasodilation in rat diaphragm microcirculation. AB - The mechanism of adenosine-induced vasodilation in rat diaphragm microcirculation was investigated using laser Doppler flowmetry. Adenosine (10(-5), 3.2 x 10(-5), and 10(-4) M), the nonselective adenosine agonist 5'-N-ethylcarboxamido-adenosine (NECA) (10(-8)-10(-7) M), the specific A(2A) agonist 2-p-(2-carboxyethyl)phenyl amino-5'-N-ethyl carboxamidoadenosine (CGS-21680) (10(-8)-10(-7) M), and the adenosine agonist with higher A(1)-receptor affinity, R-N(6) phenylisopropyladenosine (R-PIA) (10(-7), 3.2 x 10(-7), and 10(-6) M) elicited a similar degree of incremental increase of microcirculatory flow in a dose dependent manner. The ATP-dependent potassium (K(ATP)) channel blocker glibenclamide (3.2 x 10(-6) M) significantly attenuated the vasodilation effects of these agonists. Adenosine-induced vasodilation could be significantly attenuated by the nonselective adenosine antagonist 8-(p-sulfophenyl) theophylline (3 x 10(-5) M) or the selective A(2A) antagonist 4-(2-[7-amino-2-(2 furyl)[1,2, 4]triazolo[2,3-a][1,3,5]triazin-5-ylamino]ethyl) phenol (ZM-241385, 10(-6) M), but not by the selective A(1) antagonist 8-cyclopentyl-1, 3 dipropylxanthine (5 x 10(-8) M). Adenylate cyclase inhibitor N-(cis-2-phenyl cyclopentyl) azacyclotridecan-2-imine-hydrochloride (MDL-12330A, 10(-5)M) effectively suppressed the vasodilator response of adenosine and forskolin. These results suggest that adenosine-induced vasodilation in rat diaphragm microcirculation is mediated through the stimulation of A(2A) receptors, which are coupled to adenylate cyclase activation and opening of the K(ATP) channel. PMID- 11045956 TI - MRI/MRS assessment of in vivo murine cardiac metabolism, morphology, and function at physiological heart rates. AB - Transgenic mice are increasingly used to probe genetic aspects of cardiovascular pathophysiology. However, the small size and rapid rates of murine hearts make noninvasive, physiological in vivo studies of cardiac bioenergetics and contractility difficult. The aim of this report was to develop an integrated, noninvasive means of studying in vivo murine cardiac metabolism, morphology, and function under physiological conditions by adapting and modifying noninvasive cardiac magnetic resonance imaging (MRI) with image-guided (31)P magnetic resonance spectroscopy techniques used in humans to mice. Using spatially localized, noninvasive (31)P nuclear magnetic resonance spectroscopy and MRI at 4.7 T, we observe mean murine in vivo myocardial phosphocreatine-to-ATP ratios of 2.0 +/- 0.2 and left ventricular ejection fractions of 65 +/- 7% at physiological heart rates ( approximately 600 beats/min). These values in the smallest species studied to date are similar to those reported in normal humans. Although these observations do not confirm a degree of metabolic scaling with body size proposed by prior predictions, they do suggest that mice can serve, at least at this level, as a model for human cardiovascular physiology. Thus it is now possible to noninvasively study in vivo myocardial bioenergetics, morphology, and contractile function in mice under physiological conditions. PMID- 11045957 TI - Molecular distribution of volume-regulated chloride channels (ClC-2 and ClC-3) in cardiac tissues. AB - The molecular identification of cardiac chloride channels has provided probes to investigate their distribution and abundance in heart. In this study, the molecular expression and distribution of volume-regulated chloride channels ClC-2 and ClC-3 in cardiac tissues were analyzed and quantified. Total RNA was isolated from atria and ventricles of several species (dog, guinea pig, and rat) and subjected to a quantitative RT-PCR strategy. ClC-2 and ClC-3 mRNA expression were calculated relative to beta-actin expression within these same tissues. The transcriptional levels of ClC-3 mRNA were between 1.8 and 10.2% of beta-actin expression in atria and between 3.4 and 8.6% of beta-actin in ventricles (n = 3 for each tissue). The levels of ClC-2 in both atria and ventricles were significantly less than those measured for ClC-3 (n = 3; P < 0.05). ClC-2 mRNA levels were between 0.04-0.08% and 0.03-0.18% of beta-actin expression in atria and ventricles, respectively (n = 3 for each tissue). Immunoblots of atrial and ventricular wall protein extracts demonstrated ClC-2- and ClC-3-specific immunoreactivity at 97 and 85 kDa, respectively. Immunohistochemical localization in guinea pig cardiac muscle demonstrates a ubiquitous distribution of ClC-2 and ClC-3 channels in the atrial and ventricular wall. Confocal analysis detected colocalization of ClC-2 and ClC-3 in sarcolemmal membranes and distinct ClC-3 immunoreactivity in cytoplasmic regions. The molecular expression of ClC-2 and ClC-3 in cardiac tissue is consistent with the proposed role of these chloride channels in the regulation of cardiac cell volume and the modulation of cardiac electrical activity. PMID- 11045958 TI - Upregulation of p67(phox) and gp91(phox) in aortas from angiotensin II-infused mice. AB - Although NAD(P)H oxidase-derived superoxide (O(2)(-)) is increased during the development of angiotensin II (ANG II)-dependent hypertension, vascular regulation at the protein level has not been reported. We have shown that four major components of NAD(P)H oxidase are located primarily in the vascular adventitia as a primary source of vascular O(2)(-). Here we compare vascular levels of O(2)(-) and NAD(P)H oxidase in normotensive and ANG II-infused hypertensive mice and show that, after 7 days of ANG II infusion (750 microg. kg( 1). day(-1) ip) in C57B1/6 mice, systolic blood pressure was increased compared with that after sham infusion, concomitant with increased O(2)(-) in the thoracic aorta as measured using lucigenin (25 microM)-enhanced chemiluminescence. Both p67(phox) and gp91(phox) were detectable by Western blotting in aortic homogenates, and we observed increased protein levels of NAD(P)H oxidase subunits. These ANG II-induced increases were normalized by simultaneous treatment with the AT(1) receptor antagonist losartan. Moreover, the primary location of these subunits was the adventitia as detected immunohistochemically. Our results suggest that ANG II-induced increases in O(2)(-) are due to increased adventitial NAD(P)H oxidase activity, brought about by the heightened expression and interaction of its components. PMID- 11045959 TI - Expression and self-regulatory function of cardiac interleukin-6 during endotoxemia. AB - Interleukin (IL)-6 reportedly has negative inotropic and hypertrophic effects on the heart. Here, we describe endotoxin-induced IL-6 in the heart that has not previously been well characterized. An intraperitoneal injection of a bacterial lipopolysaccharide into C57BL/6 mice induced IL-6 mRNA in the heart more strongly than in any other tissue examined. Induction of mRNA for two proinflammatory cytokines, IL-1beta and tumor necrosis factor (TNF)-alpha, occurred rapidly before the induction of IL-6 mRNA and protein. Although stimulation of isolated rat neonatal myocardial cells with IL-1beta or TNF-alpha induced IL-6 mRNA in vitro, nonmyocardial heart cells produced higher levels of IL-6 mRNA upon stimulation with IL-1beta. In situ hybridization and immunohistochemical analyses localized the IL-6 expression primarily in nonmyocardial cells in vivo. Endotoxin induced expression of cardiac IL-1beta, TNF-alpha, and intercellular adhesion molecule 1 was augmented in IL-6-deficient mice compared with control mice. Thus cardiac IL-6, expressed mainly by nonmyocardial cells via IL-1beta action during endotoxemia, is likely to suppress expression of proinflammatory mediators and to regulate itself via a negative feedback mechanism. PMID- 11045960 TI - Dependence of intestinal arteriolar regulation on flow-mediated nitric oxide formation. AB - Our hypothesis was that a large fraction of resting nitric oxide (NO) formation is driven by flow-mediated mechanisms in the intestinal microvasculature of the rat. NO-sensitive microelectrodes measured the in vivo perivascular NO concentration ([NO]). Flow was increased by forcing the arterioles to perfuse additional nearby arterioles; flow was decreased by lowering the mucosal metabolic rate by reducing sodium absorption. Resting periarteriolar [NO] of large arterioles (first order; 1A) and intermediate-sized arterioles (second order; 2A) was 337 +/- 20 and 318 +/- 21 nM. The resting [NO] was higher than the dissociation constant for the NO-guanylate cyclase reaction of vascular smooth muscle; therefore, resting [NO] should be a potent dilatory signal at rest. Over flow velocity and shear rate ranges of approximately 40-180% of control, periarteriolar [NO] changed 5-8% for each 10% change in flow velocity and shear rate. The relationship of [NO] to flow velocity and shear rate demonstrated that 60-80% of resting [NO] depended on flow-mediated mechanisms. Therefore, moment-to moment regulation of [NO] at rest is an ongoing process that is highly dependent on flow-dependent mechanisms. PMID- 11045961 TI - Characterization of nifedipine-resistant calcium current in neonatal rat ventricular cardiomyocytes. AB - Calcium current was recorded from ventricular cardiomyocytes of rats at various stages of postnatal development using the whole cell patch-clamp technique. In cultured 3-day-old neonatal cells, the current carried by Ca(2+) or Ba(2+) (5 mM) was not completely inhibited by 2 microM nifedipine. A residual current was activated in the same voltage range as the L-type, nifedipine-sensitive Ca(2+) current, but its steady-state inactivation was negatively shifted by 16 mV. This nifedipine-resistant calcium current was not further inhibited by other organic calcium current antagonists such as PN200-110, verapamil, and diltiazem nor by nickel, omega-conotoxin, or tetrodotoxin. It was completely blocked by cadmium and increased by isoproterenol and forskolin. This current was >20% of total calcium current in ventricular myocytes freshly isolated from neonatal rats, and it decreased during postnatal maturation, disappearing at the adult stage. This suggests that this current could be caused by an isoform of the L-type calcium channel expressed in a way that reflects the developmental stage of the rat heart. PMID- 11045962 TI - Tyrosine kinase signaling in action potential shortening and expression of HSP72 in late preconditioning. AB - We investigated the role of tyrosine kinase (TK) signaling in the opening of the ATP-sensitive K(+) (K(ATP)) channel and 72-kDa heat shock protein (HSP72) expression during late preconditioning. Rabbits were subjected to surgical operation (sham) or were preconditioned (PC) with four cycles of 5 min of ischemia and 10 min of reperfusion. Twenty-four hours later, animals were subjected to 30 min of ischemia and 180 min of reperfusion. Genistein (1 mg/kg ip) was used to block the receptor TK. Six groups were studied: control, sham, genistein-sham, PC, genistein-PC, and vehicle-PC group (1% dimethyl sulfoxide). Genistein or vehicle was given 30 min before the surgical procedure. Genistein pretreatment decreased the expression of HSP72 in PC hearts and suppressed action potential duration shortening during ischemia in sham and PC groups. Infarct size (%risk area) was reduced in the PC (11.6 +/- 1.0%) and vehicle-PC (19.3 +/- 2.0%) compared with the control (40.0 +/- 3.8%) or sham (46.0 +/- 2.0%) groups (P < 0.05). Genistein pretreatment increased infarct size to 46.4 +/- 4.1% in the PC hearts. We conclude that TK signaling is involved in K(ATP) channel opening and HSP72 expression during late PC. PMID- 11045963 TI - Acute effects of 17beta-estradiol on ventricular and vascular hemodynamics in postmenopausal women. AB - Because premenopausal women have lower cardiovascular morbidity than postmenopausal women, it has been proposed that estrogen may have a protective role. Estrogen is involved in smooth muscle relaxation both through its specific receptor as well as through calcium channel blockade. This study examined the acute effect of estradiol on invasive cardiovascular hemodynamics in 18 postmenopausal women (age 62.6 +/- 7.6 years, means +/- SD). The effect of estradiol on left ventricular chamber performance was studied in 9 women using simultaneous left ventricular pressure-volume recordings. In a further group of 9 women, the acute effect of estradiol on arterial function was assessed using input impedance (derived from simultaneous aortic pressure and flow recordings), pressure waveform analysis, and pulse wave velocity. After 2 mg micronized 17beta estradiol was administered, serum estradiol levels increased from 50.9 +/- 21.9 to 3,190 +/- 2,216 pmol/l, P < 0.0001. There was no effect of estradiol on either left ventricular inotropic or lusitropic function. There was no acute effect of estradiol on arterial impedance, reflection coefficient, augmentation index, or pulse wave velocity. There was a trend to decreased heart rate and cardiac output in both groups of 9 women. Because heart rate and cardiac output were common to both hemodynamic data sets, results for these parameters were pooled. Across all 18 women, there was a small but significant decrease in heart rate (69.2 +/- 10.4 vs. 67.2 +/- 9.9 beats/min, P = 0.02), as well as a significant decrease in cardiac output (4.82 +/- 1.77 vs. 4.17 +/- 1.56 l/min, P = 0.002). Despite achieving supraphysiological serum levels, this study found no significant effect of acute 17beta-estradiol on ventricular or large artery function. PMID- 11045964 TI - Local and remote arteriolar dilations initiated by skeletal muscle contraction. AB - To investigate the relationship between skeletal muscle metabolism and arteriolar dilations in the region local to contracting muscle fibers as well as dilations at remote arteriolar regions upstream, we used a microelectrode on cremaster muscle of anesthetized hamsters to stimulate four to five muscle fibers lying approximately perpendicular to and overlapping a transverse arteriole. Before, during, and after muscle contraction, we measured the diameter of the arteriole at the site of muscle fiber overlap (local) and at a remote site approximately 1,000 microm upstream. Two minutes of 2-, 4-, or 8-Hz stimulation (5-10 V, 0.4-ms duration) produced a significant dilation locally (8.2 +/- 2.0-, 22.5 +/- 2.4-, and 30.9 +/- 2.1-microm increase, respectively) and at the remote site (4.2 +/- 0.8, 11.0 +/- 1.1, and 18.9 +/- 2.7 microm, respectively). Muscle contraction at 4 Hz initiated a remote dilation that was unaffected by 15-min micropipette application of either 2 microM tetrodotoxin, 0.07% halothane, or 40 microM 18 beta-glycyrrhetinic acid between the local and upstream site. Therefore, at the arteriolar level, muscle contraction initiates a robust remote dilation that does not appear to be transmitted via perivascular nerves or gap junctions. PMID- 11045965 TI - Microvascular flow and tissue PO(2) in skeletal muscle of chronic reduced renal mass hypertensive rats. AB - This study determined whether arteriolar blood flow, capillary red blood cell (RBC) velocity, capillary hematocrit (Hct(cap)), and tissue PO(2) are altered in cremaster muscles of rats with chronic reduced renal mass hypertension (RRM-HT) relative to normotensive rats on high- or low-salt (NT-HS vs. NT-LS) diet. The blood flow in first- through third-order arterioles was not different between NT and HT rats, either at rest or during maximal relaxation of the vessels with 10( 4) M adenosine. Capillary RBC velocity was similar between the groups at rest but was elevated in RRM-HT and NT-HS rats during adenosine superfusion. Hct(cap) was reduced at rest in RRM-HT and NT-HS rats compared with NT-LS and was reduced in RRM-HT rats during adenosine-induced dilation. Tissue PO(2) was reduced in RRM-HT and NT-HS rats compared with NT-LS rats during control conditions and was lower in RRM-HT than in NT-LS rats during adenosine-induced dilation. These results indicate that both RRM-HT and chronic exposure of normotensive rats to a high salt diet lead to reduced tissue oxygenation, despite the maintenance of normal arteriolar blood flow. PMID- 11045966 TI - Na(+) current contribution to the diastolic depolarization in newborn rabbit SA node cells. AB - Isolated newborn, but not adult, rabbit sinoatrial node (SAN) cells exhibit spontaneous activity that (unlike adult) are highly sensitive to the Na(+) current (I(Na)) blocker TTX. To investigate this TTX action on automaticity, cells were voltage clamped with ramp depolarizations mimicking the pacemaker phase of spontaneous cells (-60 to -20 mV, 35 mV/s). Ramps elicited a TTX sensitive current in newborn (peak density 0.89 +/- 0.14 pA/pF, n = 24) but not adult (n = 5) cells. When depolarizing ramps were preceded by steplike depolarizations to mimic action potentials, ramp current decreased 54.6 +/- 8.0% (n = 3) but was not abolished. Additional experiments demonstrated that ramp current amplitude depended on the slope of the ramp and that TTX did not alter steady-state holding current at pacemaker potentials. This excluded a steady state Na(+) window component and suggested a kinetic basis, which was investigated by measuring TTX-sensitive I(Na) during long step depolarizations. I(Na) exhibited a slow but complete inactivation time course at pacemaker voltages (tau = 33.9 +/- 3.9 ms at -50 mV), consistent with the rate-dependent ramp data. The data indicate that owing to slow inactivation of I(Na) at diastolic potentials, a small TTX-sensitive current flows during the diastolic depolarization in neonatal pacemaker myocytes. PMID- 11045967 TI - Hypoxia-induced alterations in Ca(2+) mobilization in brain microvascular endothelial cells. AB - To investigate the possible cellular mechanisms of the ischemia-induced impairments of cerebral microcirculation, we investigated the effects of hypoxia/reoxygenation on the intracellular Ca(2+) concentration ([Ca(2+)](i)) in bovine brain microvascular endothelial cells (BBEC). In the cells kept in normal air, ATP elicited Ca(2+) oscillations in a concentration-dependent manner. When the cells were exposed to hypoxia for 6 h and subsequent reoxygenation for 45 min, the basal level of [Ca(2+)](i) was increased from 32.4 to 63.3 nM, and ATP did not induce Ca(2+) oscillations. Hypoxia/reoxygenation also inhibited capacitative Ca(2+) entry (CCE), which was evoked by thapsigargin (Delta[Ca(2+)](i-CCE): control, 62.3 +/- 3.1 nM; hypoxia/reoxygenation, 17.0 +/- 1.8 nM). The impairments of Ca(2+) oscillations and CCE, but not basal [Ca(2+)](i), were restored by superoxide dismutase and the inhibitors of mitochondrial electron transport, rotenone and thenoyltrifluoroacetone (TTFA). By using a superoxide anion (O(2)(-))-sensitive luciferin derivative MCLA, we confirmed that the production of O(2)(-) was induced by hypoxia/reoxygenation and was prevented by rotenone and TTFA. These results indicate that hypoxia/reoxygenation generates O(2)(-) at mitochondria and impairs some Ca(2+) mobilizing properties in BBEC. PMID- 11045968 TI - Effect of sildenafil on coronary active and reactive hyperemia. AB - Sildenafil, a selective inhibitor of phosphodiesterase type 5, produces relaxation of isolated epicardial coronary artery segments by causing accumulation of cGMP. Because shear-induced nitric oxide-dependent vasodilation is mediated by cGMP, this study was performed to determine whether sildenafil would augment the coronary resistance vessel dilation that occurs during the high flow states of exercise or reactive hyperemia. In chronically instrumented dogs, sildenafil (2 mg/kg per os) augmented the vasodilator response to acetylcholine, with a leftward shift of the dose-response curve relating coronary flow to acetylcholine dose. Sildenafil caused a 6. 7 +/- 2.1 mmHg decrease of mean aortic pressure, which was similar at rest and during treadmill exercise (P < 0.05), with no change of heart rate, left ventricular (LV) systolic pressure, or LV maximal first time derivative of LV pressure. Sildenafil tended to increase myocardial blood flow at rest and during exercise (mean increase = 14 +/- 3%; P < 0.05 by ANOVA), but this was associated with a significant decrease in hemoglobin, so that the relationship between myocardial oxygen consumption and oxygen delivery to the myocardium (myocardial blood flow x arterial O(2) content) was unchanged. Furthermore, sildenafil did not alter coronary venous PO(2), indicating that the coupling between myocardial blood flow and myocardial oxygen demands was not altered. In addition, sildenafil did not alter the peak coronary flow rate, debt repayment, or duration of reactive hyperemia that followed a 10-s coronary occlusion. The findings suggest that cGMP-mediated resistance vessel dilation contributes little to the increase in myocardial flow that occurs during exercise or reactive hyperemia. PMID- 11045969 TI - Hemodynamic changes in apolipoprotein E-knockout mice. AB - Apolipoprotein E-knockout (ApoE-KO) mice develop advanced atherosclerotic lesions by 1 yr of age and have been well characterized pathologically and morphologically, but little is known regarding their cardiovascular physiology and hemodynamics. We used noninvasive Doppler ultrasound to measure aortic and mitral blood velocity and aortic pulse-wave velocity in 13-mo-old ApoE-KO and wild-type (WT) mice anesthetized with isoflurane. In other mice from the same colony, we measured systolic blood pressure, body weight, heart weight, cholesterol, and hematocrit. Heart rate and blood pressure were comparable (P = not significant) between ApoE-KO and WT mice, but significant decreases (P < 0.001) were found in body weight (-22%) and hematocrit (-11%), and significant increases were found in heart weight (+23%), aortic velocity (+60%), mitral velocity (+81%) (all P < 0.001), and pulse-wave velocity (+13%, P < 0.05). We also found inflections in the aortic arch velocity signal consistent with enhanced peripheral wave reflection. Thus ApoE-KO mice have phenotypic alterations in indexes of peripheral vascular resistance and compliance and significantly elevated cardiac outflow velocities and heart weight-to-body weight ratios. PMID- 11045970 TI - Changes in venous return parameters associated with univentricular Fontan circulations. AB - To clarify the physiology of venous return (Q(vr)) in Fontan circulations, venous return conductance (G(vr)) and mean circulatory filling pressure (P(mcf)) were determined in pentobarbital sodium-anesthetized pigs. Relationships between Q(vr) and right (biventricular, n = 8) or left (Fontan, n = 8) filling pressures are described by straight lines with significant correlation coefficients. Estimated P(mcf) values were correlated with observed P(mcf) values in either circulations (P 36%) IS developed lung congestion, cardiac hypertrophy, left ventricular (LV) dilatation, elevated LV end-diastolic pressure (LVEDP), and suppressed maximal rate of increase of LV pressure (LV dP/dt(max)) and fractional shortening (FS). Whereas changes in organ weights and echo parameters were similar to those in infarcted WT groups, TG mice had significantly higher levels of LV contractility in both moderate (dP/dt(max) 4,862 +/- 133 vs. 3,694 +/- 191 mmHg/s) and large IS groups (dP/dt(max) 4,556 +/- 252 vs. 3,145 +/- 312 mmHg/s, both P < 0.01). Incidence of pleural effusion (36% vs. 85%, P < 0.05) and LVEDP levels (6 +/- 0.3 vs. 9 +/- 0.8 mmHg, P < 0.05) were also lower in TG than in WT mice with large IS. Thus beta(2)-AR overexpression preserved LV contractility following MI without adverse consequence. PMID- 11045984 TI - Normal values of regional left ventricular endocardial motion: multicenter color kinesis study. AB - Our goal was to establish normal values for quantitative color kinesis indexes of left ventricular (LV) wall motion over a wide range of ages, which are required for objective diagnosis of regional systolic and diastolic dysfunction. Color encoded images were obtained in 194 normal subjects (95 males, 99 females, age 2 mo to 79 yr) in four standard views. Quantitative indexes of magnitude and timing of systolic and diastolic function were studied for age- and gender-related differences. Normal limits of all ejection and filling indexes were in a narrow range (< or =25% of the mean), with no major gender-related differences. Despite invariable ejection fractions, both peak filling and ejection rates decreased with age (30 and 20%, correspondingly) with a concomitant increase in mean filling and ejection times, resulting in five- and twofold increases in the late to early filling and ejection ratios, correspondingly. Diastolic asynchrony increased with age (from 4.7 +/- 2.0 to 6.4 +/- 3.2 from the 2nd to 7th decade). The normal values of color kinesis indexes should allow objective detection of regional LV systolic and diastolic dysfunction. PMID- 11045985 TI - Cyclic strain modulates resistance to oxidant stress by increasing G6PDH expression in smooth muscle cells. AB - Vascular smooth muscle cells (VSMC) may be subjected to mechanical forces, such as cyclic strain, that promote the formation of reactive oxygen species (ROS). We hypothesized that VSMC modulate this adverse milieu by increasing the expression of glucose-6-phosphate dehydrogenase (G6PDH) to maintain or restore intracellular glutathione (GSH) levels. Cyclic strain increased superoxide formation, which resulted in diminished GSH because of an increase in oxidized glutathione formation; there was also an increase in glutathione peroxidase and glutathione reductase activities. G6PDH activity and protein expression were enhanced concomitant with decreases in GSH levels and remained elevated until intracellular GSH levels were restored. To confirm the role of G6PDH in repleting GSH stores, we inhibited G6PDH activity with DHEA or inhibited enzyme expression with an antisense oligodeoxynucleotide. Diminished G6PDH activity or expression was associated with persistently depleted GSH levels and inhibition of the cyclic strain-mediated increase in glutathione reductase activity. These observations demonstrate that cyclic strain promotes oxidant stress in VSMC, which, in turn, induces G6PDH expression. When G6PDH is inhibited, GSH levels are not restored because of impaired glutathione reductase activity. These data suggest that G6PDH is a critical determinant of the response to oxidant stress in VSMC. PMID- 11045986 TI - Effects of whole body heating on dynamic baroreflex regulation of heart rate in humans. AB - The purpose of this project was to identify whether dynamic baroreflex regulation of heart rate (HR) is altered during whole body heating. In 14 subjects, dynamic baroreflex regulation of HR was assessed using transfer function analysis. In normothermic and heat-stressed conditions, each subject breathed at a fixed rate (0. 25 Hz) while beat-by-beat HR and systolic blood pressure (SBP) were obtained. Whole body heating significantly increased sublingual temperature, HR, and forearm skin blood flow. Spectral analysis of HR and SBP revealed that the heat stress significantly reduced HR and SBP variability within the high-frequency range (0.2-0.3 Hz), reduced SBP variability within the low-frequency range (0.03 0.15 Hz), and increased the ratio of low- to high-frequency HR variability (all P < 0.01). Transfer function gain analysis showed that the heat stress reduced dynamic baroreflex regulation of HR within the high-frequency range (from 1.04 +/ 0.06 to 0.54 +/- 0.6 beats. min(-1). mmHg(-1); P < 0.001) without significantly affecting the gain in the low-frequency range (P = 0.63). These data suggest that whole body heating reduced high-frequency dynamic baroreflex regulation of HR associated with spontaneous changes in blood pressure. Reduced vagal baroreflex regulation of HR may contribute to reduced orthostatic tolerance known to occur in humans during heat stress. PMID- 11045987 TI - Effects of microtubule disruption on force, velocity, stiffness and [Ca(2+)](i) in porcine coronary arteries. AB - Force generated by smooth muscle cells is believed to result from the interaction of actin and myosin filaments and is regulated through phosphorylation of the myosin regulatory light chain (LC(20)). The role of other cytoskeleton filaments, such as microtubules and intermediate filaments, in determining the mechanical output of smooth muscle is unclear. In cultured fibroblasts, microtubule disruption results in large increases in force similar to contractions associated with LC(20) phosphorylation (15). One hypothesis, the "tensegrity" or "push-pull" model, attributes this increase in force to the disruption of microtubules functioning as rigid struts to resist force generated by actin-myosin interaction (9). In porcine coronary arteries, the disruption of microtubules by nocodazole (11 microM) also elicited moderate but significant increases in isometric force (10-40% of a KCl contracture), which could be blocked or reversed by taxol (a microtubule stabilizer). We tested whether this nocodazole-induced force was accompanied by changes in coronary artery stiffness or unloaded shortening velocity, parameters likely to be highly sensitive to microtubule resistance elements. Few changes were seen, ruling out push-pull mechanisms for the increase in force by nocodazole. In contrast, the intracellular calcium concentration, measured by fura 2 in the intact artery, was increased by nocodazole in parallel with force, and this was inhibited and/or reversed by taxol. Our results indicate that microtubules do not significantly contribute to vascular smooth muscle mechanical characteristics but, importantly, may play a role in modulation of Ca(2+) signal transduction. PMID- 11045988 TI - Altered beta-adrenergic signal transduction in nonfailing hypertrophied myocytes from Dahl salt-sensitive rats. AB - Desensitization of the beta-adrenergic receptor (beta-AR) response is well documented in hypertrophied hearts. We investigated whether beta-AR desensitization is also present at the cellular level in hypertrophied myocardium, as well as the physiological role of inhibitory G (G(i)) proteins and the L-type Ca(2+) channel in mediating beta-AR desensitization. Left ventricular (LV) myocytes were isolated from hypertrophied hearts of hypertensive Dahl salt sensitive (DS) rats and nonhypertrophied hearts of normotensive salt-resistant (DR) rats. Cells were paced at a rate of 300 beats/min at 37 degrees C, and myocyte contractility and intracellular Ca(2+) concentration ([Ca(2+)](i)) were simultaneously measured. In response to increasing concentrations of isoproterenol, DR myocytes displayed a dose-dependent augmentation of cell shortening and the [Ca(2+)](i) transient amplitude, whereas hypertrophied DS myocytes had a blunted response of both cell shortening and the [Ca(2+)](i) transient amplitude. Interestingly, inhibition of G(i) proteins did not restore beta-AR desensitization in DS myocytes. The responses to increases in extracellular Ca(2+) and an L-type Ca(2+) channel agonist were also similar in both DS and DR myocytes. Isoproterenol-stimulated adenylyl cyclase activity, however, was blunted in hypertrophied myocytes. We concluded that compensated ventricular hypertrophy results in a blunted contractile response to beta-AR stimulation, which is present at the cellular level and independent of alterations in inhibitory G proteins and the L-type Ca(2+) channel. PMID- 11045989 TI - NO is involved in MCh-induced accentuated antagonism via type II PDE in the canine blood-perfused SA node. AB - The possible role of type II (cGMP-stimulated cAMP hydrolysis) phosphodiesterase (PDE) in the accentuated antagonism of muscarinic effects on heart rate during beta-stimulation via endogenous nitric oxide (NO) was evaluated. The canine isolated sinoatrial node preparation was cross circulated with arterial blood of a support dog. The sinoatrial rate of the preparation was 96 +/- 5 beats/min (n = 16) at control. Methacholine (MCh; 0.01-1 microg) injected into the right coronary artery in a bolus fashion caused dose-dependent decreases in sinoatrial rate. Under an intra-arterial infusion of isoproterenol (1 microM), resulting in approximately 50% increase in sinoatrial rate, MCh-induced decreases were markedly augmented from -18 +/- 3% to -44 +/- 4% at 0.3 mg of MCh. When N(G) nitro-L-arginine methyl ester (100 microM) or N(G)-monomethyl-L-arginine (100 microM) were continuously infused, the augmented MCh-induced decreases in sinoatrial rate were significantly suppressed (-29 +/- 3% or -25 +/- 3%, respectively, P < 0.01). Pretreatment with either 3-isobutyl-1-methylxanthine (IBMX; 20 microM), a non-selective PDE inhibitor, or amrinone (20 microM), a selective type III (cGMP inhibited cAMP hydrolysis) PDE inhibitor, doubled the isoproterenol-induced increase in the sinoatrial rate. However, the augmented MCh induced decreases in sinoatrial rate were significantly depressed by IBMX (from 23 +/- 5% to -14 +/- 1%, P < 0.01) but not by amrinone (to -20 +/- 3%). These results suggest that MCh-induced accentuated antagonism in the sinoatrial node pacemaker activity can be modulated by endogenous NO via an activation of the type II cyclic GMP-stimulated cAMP PDE. PMID- 11045990 TI - Active and passive stresses in the myocardium. AB - A mathematical approach that can be used to calculate the passive stress in the ventricular wall is presented. The active fiber stress (force/unit area) generated by the muscular fibers in the ventricular wall is expressed by means of body force (force/unit volume of the myocardium). It is shown that the total intramyocardial passive stress induced in the passive medium of the myocardium can be expressed as the sum of a passive stress induced by the left ventricular pressure and a passive stress induced by the active fiber stress. Applications to experimental data published in the literature are given. New results are presented that show the relation among those two components of the intramyocardial passive stress. New relations between the intramyocardial passive stress, the slope (elastance) of the pressure-volume relation, and the residual volume are also derived. The results obtained give a better understanding of some aspects of the mechanics of cardiac contraction and can provide a more detailed interpretation of clinical conditions. PMID- 11045991 TI - Mediators of the mitogenic action of human V(1) vascular vasopressin receptors. AB - Arginine vasopressin (AVP) activation of V(1) vascular receptors (V(1)Rs) stimulates cell growth and proliferation in different tissues via cellular signaling pathways that remain to be identified. To explore the intracellular mediators of the mitogenic action of V(1)R, Chinese hamster ovary (CHO) cells were stably transfected with the human V(1)R cDNA clone we isolated previously. We assessed AVP effects on kinase activation (immunoblotting with phosphospecific antibodies), DNA synthesis (tritiated thymidine uptake), cell cycle progression (flow cytometry analysis after nuclear labeling with propidium iodide), and cell proliferation (conversion of the colorimetric reagent MTS) in the presence or absence of various pathway inhibitors. AVP stimulation of V(1)Rs leads to the phosphorylation of several kinases, an increase in DNA synthesis, a progression through the S and G(2)-M phases of the cell cycle, and an increase in cell proliferation. The mediators of the mitogenic action of V(1)R activation included calcium mobilization, coupling to a G(q) protein, and the simultaneous and parallel activation of several kinases, mainly calcium/calmodulin-dependent kinase II, phosphatidylinositol 3 kinase, protein kinase C, and p42/p44 mitogen activated protein kinase. PMID- 11045992 TI - T-type calcium currents in rat cardiomyocytes during postnatal development: contribution to hormone secretion. AB - T-type Ca(2+) channels have been suggested to play a role in cardiac automaticity, cell growth, and cardiovascular remodeling. Although three genes encoding for a T-type Ca(2+) channel have been identified, the nature of the isoform(s) supporting the cardiac T-type Ca(2+) current (I(Ca,T)) has not yet been determined. We describe the postnatal evolution of I(Ca,T) density in freshly dissociated rat atrial and ventricular myocytes and its functional properties at peak current density in young atrial myocytes. I(Ca,T) displays a classical low activation threshold, rapid inactivation kinetics, negative steady state inactivation, slow deactivation, and the presence of a window current. Interestingly, I(Ca,T) is poorly sensitive to Ni(2+) and insensitive to R-type current toxin SNX-482. RT-PCR experiments and comparison of functional properties with recombinant Ca(2+) channel subtypes suggest that neonatal I(Ca,T) is related to the alpha(1G)-subunit. Atrial natriuretic factor (ANF) secretion was measured using peptide radioimmunoassays in atrial tissue. Pharmacological dissection of ANF secretion indicates an important contribution of I(Ca,T) to Ca(2+) signaling during the neonatal period. PMID- 11045993 TI - Responses of neurons in rostral ventrolateral medulla to activation of cardiac receptors in rats. AB - Ischemic stimulation of cardiac receptors reflexly excites the cardiovascular system. However, the supraspinal mechanisms involved in this reflex are not well defined. This study examined the responses of barosensitive neurons in the rostral ventrolateral medulla (RVLM) to stimulation of cardiac receptors and the afferent pathways involved in these responses. Single-unit activity of RVLM neurons was recorded in alpha-chloralose-anesthetized rats. Cardiac receptors were stimulated by epicardial application of 10 microg/ml of bradykinin (BK). Barosensitive neurons were silenced by stimulation of baroreceptors. Application of BK increased the mean arterial pressure from 65.2 +/- 1.9 to 89.3 +/- 2.9 mmHg and excited RVLM barosensitive neurons from 6.2 +/- 0.7 to 10.7 +/- 0.9 impulses/s (P < 0.05, n = 40). BK had no effect on 21 nonbarosensitive neurons. Blockade of stellate ganglia abolished the response of barosensitive neurons to BK. Cervical vagotomy significantly increased the baseline discharges of RVLM barosensitive neurons but had no effect on their responses to BK. Thus this study indicates that stimulation of cardiac receptors selectively activates RVLM barosensitive neurons through sympathetic afferent pathways. This information suggests that the RVLM barosensitive neurons are likely involved in the sympathetic control of circulation during myocardial ischemia. PMID- 11045994 TI - Assessing baroreflex gain from spontaneous variability in conscious dogs: role of causality and respiration. AB - A double exogenous autoregressive (XXAR) causal parametric model was used to estimate the baroreflex gain (alpha(XXAR)) from spontaneous R-R interval and systolic arterial pressure (SAP) variabilities in conscious dogs. This model takes into account 1) effects of current and past SAP variations on the R-R interval (i.e., baroreflex-mediated influences), 2) specific perturbations affecting R-R interval independently of baroreflex circuit (e.g., rhythmic neural inputs modulating R-R interval independently of SAP at frequencies slower than respiration), and 3) influences of respiration-related sources acting independently of baroreflex pathway (e.g., rhythmic neural inputs modulating R-R interval independently of SAP at respiratory rate, including the effect of stimulation of low-pressure receptors). Under control conditions, alpha(XXAR) = 14.7 +/- 7.2 ms/mmHg. It decreases after nitroglycerine infusion and coronary artery occlusion, even though the decrease is significant only after nitroglycerine, and it is completely abolished by total arterial baroreceptor denervation. Moreover, alpha(XXAR) is comparable to or significantly smaller than (depending on the experimental condition) the baroreflex gains derived from sequence, power spectrum [at low frequency (LF) and high frequency (HF)], and cross-spectrum (at LF and HF) analyses and from less complex causal parametric models, thus demonstrating that simpler estimates may be biased by the contemporaneous presence of regulatory mechanisms other than baroreflex mechanisms. PMID- 11045995 TI - Myofilament lattice spacing as a function of sarcomere length in isolated rat myocardium. AB - The Frank-Starling relationship of the heart has, as its molecular basis, an increase in the activation of myofibrils by calcium as the sarcomere length increases. It has been suggested that this phenomenon may be due to myofilaments moving closer together at longer lengths, thereby enhancing the probability of favorable acto-myosin interaction, resulting in increased calcium sensitivity. Accordingly, we have developed an apparatus so as to obtain accurate measurements of myocardial interfilament spacing (by synchrotron X-ray diffraction) as a function of sarcomere length (by video microscopy) over the working range of the heart, using skinned as well as intact rat trabeculas as model systems. In both these systems, lattice spacing decreased significantly as sarcomere length was increased. Furthermore, lattice spacing in the intact muscle was significantly smaller than that in the skinned muscle at all sarcomere lengths studied. These observations are consistent with the hypothesis that lattice spacing underlies length-dependent activation in the myocardium. PMID- 11045996 TI - K(ATP) channels mediate the beneficial effects of chronic ethanol ingestion. AB - Chronic ingestion of low doses of ethanol protects the myocardium from ischemic injury by activating adenosine receptors and protein kinase C. We tested the hypothesis that ATP-dependent potassium (K(ATP)) channels mediate these beneficial effects. Dogs were fed with ethanol (1.5 g/kg) or water mixed with dry food twice per day for 12 wk. After they were acutely instrumented for measurement of hemodynamics, dogs received saline (vehicle) or glyburide (0.1 mg/kg iv) and were subjected to 60 min of coronary artery occlusion followed by 3 h of reperfusion. Infarct size (through triphenyltetrazolium chloride staining) was significantly (P < 0.05) reduced to 14 +/- 1% of the left ventricular area at risk in ethanol-pretreated dogs compared with controls (25 +/- 2%). Glyburide alone did not affect infarct size (25 +/- 3%) but abolished the protective effects of ethanol pretreatment (28 +/- 3%). No differences in hemodynamics or coronary collateral blood flow (through radioactive microspheres) were observed among groups. The results indicate that K(ATP) channels mediate the protective effects of chronic consumption of ethanol. PMID- 11045997 TI - Cutting edge: TGF-beta inhibits Th type 2 development through inhibition of GATA 3 expression. AB - TGF-beta is an important immunomodulatory cytokine that can inhibit differentiation of effector T cells. In this report, we address the molecular mechanisms through which TGF-beta inhibits differentiation of CD4(+) cells into Th type 2 cells. We demonstrate that TGF-beta inhibits GATA-3 expression in developing Th cells. We also show that inhibition of GATA-3 expression by TGF beta is a major mechanism of inhibition of Th2 differentiation by TGF-beta as ectopic expression of GATA-3 in developing T cells overcomes the ability of TGF beta to inhibit Th2 differentiation. TGF-beta likely inhibits GATA-3 expression at the transcriptional level and does so without interfering with IL-4 signaling. PMID- 11045998 TI - Cutting edge: the mucosal adjuvant cholera toxin redirects vaccine proteins into olfactory tissues. AB - We tested the notion that the mucosal adjuvant cholera toxin (CT) could target, in addition to nasal-associated lymphoreticular tissues, the olfactory nerves/epithelium (ON/E) and olfactory bulbs (OBs) when given intranasally. Radiolabeled CT ((125)I-CT) or CT-B subunit ((125)I-CT-B), when given intranasally to mice, entered the ON/E and OB and persisted for 6 days; however, neither molecule was present in nasal-associated lymphoreticular tissues beyond 24 h. This uptake into olfactory regions was monosialoganglioside (GM1) dependent. Intranasal vaccination with (125)I-tetanus toxoid together with unlabeled CT as adjuvant resulted in uptake into the ON/E but not the OB, whereas (125)I-tetanus toxoid alone did not penetrate into the CNS. We conclude that GM1 binding molecules like CT target the ON/E and are retrograde transported to the OB and may promote uptake of vaccine proteins into olfactory neurons. This raises concerns about the role of GM1-binding molecules that target neuronal tissues in mucosal immunity. PMID- 11045999 TI - Cutting edge: anti-CD154 therapeutic antibodies induce infectious transplantation tolerance. AB - Nondepleting anti-CD154 (CD40 ligand) mAbs have proven effective in inducing transplantation tolerance in rodents and primates. In the induction phase, anti CD154 Ab therapy is known to enhance apoptosis of Ag reactive T cells. However, this may not be the sole explanation for tolerance, as we show in this study that tolerance is maintained through a dominant regulatory mechanism which, like tolerance induced with CD4 Abs, manifests as infectious tolerance. Therefore, tolerance induced with anti-CD154 Abs involves not only the deletion of potentially aggressive T cells, but also a contagious spread of tolerance to new cohorts of graft-reactive T cells as they arise. PMID- 11046000 TI - Cutting edge: selective IL-18 requirements for induction of compartmental IFN gamma responses during viral infection. AB - Optimal protective effects for defense against infection require orchestration of immune responses spanning multiple host compartments and divergent local regulation at particular sites. During murine cytomegalovirus infections known to target spleen and liver, IL-12-induced IFN-gamma from NK cells is crucial for resistance. However, the roles for IL-18 and/or IL-12 in regulating hepatic IFN gamma responses, as compared with systemic or splenic responses, have not been defined. In this report, mice genetically deficient in either IL-18 or IL-12p35 exhibited up to 95% reductions in systemic and splenic IFN-gamma responses. Surprisingly, IFN-gamma responses were preserved in the livers of IL-18 deficient, but not IL-12p35-deficient, mice. Cytokine requirements for host survival also differed. Under conditions where mice lacking IL-12p35 exhibited 100% mortality, those lacking IL-18 survived. Taken together, our results delineate contrasting compartmental requirements for IL-18 and suggest that preservation of local, hepatic IFN-gamma production is critical for host defense during murine cytomegalovirus challenge. PMID- 11046001 TI - Cutting edge: resistance to apoptosis and continuous proliferation of dendritic cells deficient for TNF receptor-1. AB - The individual roles of the two TNFRs on dendritic cells (DC) are poorly understood. Investigating bone marrow-derived DC from TNFR-deficient mice, we found that cultures from TNFR1(-/-) mice continue to form proliferating clusters for 6-9 mo. In contrast, DC derived from wild-type, TNFR2(-/-), or TNFR1/2(-/-) mice survived for only 3-4 wk. DC obtained from these TNFR1(-/-) long term cultures (LTC) mice show an unusual mixed immature/mature phenotype. The continuous proliferation of the LTC is GM-CSF dependent and correlates with decreased protein levels of the cyclin-dependent kinase inhibitors p27(KIP1) and p21(CIP1). Prolonged survival of TNFR1(-/-) DC appears to be independent from NF kappaB and Bcl-2 pathways and is rather enabled by the down-regulation of CD95, resulting in the resistance to CD95 ligand-induced apoptosis. These data point to proapoptotic signals mediated via TNFR1 and antiapoptotic signals mediated via TNFR2 in DC. PMID- 11046002 TI - Cutting edge: CD1d deficiency impairs murine host defense against the spirochete, Borrelia burgdorferi. AB - CD1 molecules can present microbial lipid Ag to T cells, suggesting that they participate in host defense against pathogens. In this study, we examined the role of CD1d in resistance to infection with the Lyme disease spirochete, Borrelia burgdorferi (Bb), an organism with proinflammatory lipid Ag. Bb infection of CD1d-deficient (CD1d(-/-)) mouse strains normally resistant to this pathogen resulted in arthritis. Pathology correlated with an increased prevalence of spirochete DNA in tissues and enhanced production of Bb-specific IgG, including IgG to Ag rapidly down-modulated on spirochetes in vivo. CD1d(-/-) mice exhibited high-titer Bb-specific IgG2a, an isotype commonly induced in disease susceptible mice but not in the disease-resistant control mice in this study. These results show that CD1d deficiency impairs host resistance to a spirochete pathogen, and are the first example of a mutation that imparts Bb-resistant mice with the Ab and disease profile of a susceptible mouse strain. PMID- 11046003 TI - The ability of B cells and dendritic cells to present antigen increases during ontogeny. AB - The immune response to polysaccharide (PS) Ags in mice is delayed during ontogeny even when administered in a thymus-dependent (TD) form. In this study, Neisseria meningitidis group C PS-tetanus toxoid conjugate (MCPS-TT) vaccine was used to examine whether the delay in the development of Ab responses to TD PS conjugate vaccines in neonatal mice is due to defective Ag presentation. The results show that B cells and dendritic cells (DC) from 3- and 7-day-old mice were severely defective in presenting TT and MCPS-TT to Ag-specific T cell clones. The ability of these cells to present Ag reaches adult levels by 4 wk. The development of anti-MCPS and anti-TT Abs in neonatal mice parallels the functional ability of their APC to present Ag. DC from neonatal mice expressed very low levels of MHC class II, costimulatory molecules B7.1, B7.2, and CD11c but high levels of monocyte-specific markers F4/80 and CD11b and granulocyte marker, Ly6G. Significant changes in the expression of these markers were observed as the age of the mice increased. MHC class II, B7.1 and B7.2, and CD11c all increased with age, reaching adult levels between 3 and 4 wk, concurrent with the function of APC. These results demonstrate that one reason neonates fail to produce high titers of anti-PS Abs even when immunized in a TD form is that their B cells and DC are not fully functional. PMID- 11046004 TI - TNF receptor 2-deficient CD8 T cells are resistant to Fas/Fas ligand-induced cell death. AB - Apoptotic cell death plays a fundamental role in the maintenance of tissue homeostasis in complex biological systems. It is also a major mechanism for keeping immune reactions in check. Members of the TNF family of receptors and cytokines are implicated in the regulation of apoptotic signals that shape the immune system. In this study, we have examined the role of three members of the TNFR family, Fas (CD95), TNFR1 (p55), and TNFR2 (p75), in inducing cell death in Con A-activated CD4 and CD8 T cells. It was found that Con A-activated p55(-/-) CD4 or CD8 T cells were highly resistant to TNF-induced cell death. By contrast, although activated p75(-/-) CD4 or CD8 T cells were killed by TNF, they were more resistant to TNF-induced killing when compared with p75(+/+) cells, particularly at higher concentrations of TNF. We also determined whether activated p55(-/-) and p75(-/-) T cells differ in their sensitivity to cell death induced by TCR cross-linking. We found that activated p55(-/-) CD4 or CD8 T cells were equally susceptible to TCR-induced cell death. More interestingly, the loss of the p75 receptor conferred resistance to TCR-induced death in activated CD8, but not CD4 T cells. This resistance to TCR-induced death in activated p75(-/-) CD8 T cells correlated with the resistance of these cells to Fas/Fas ligand-induced cell death. PMID- 11046006 TI - Changing patterns of dominant TCR usage with maturation of an EBV-specific cytotoxic T cell response. AB - Infection with EBV provides a unique opportunity to follow the human CD8(+) T cell response to a persistent, genetically stable agent from the primary phase, as seen in infectious mononucleosis (IM) patients, into long-term memory. This study focuses on the response to an immunodominant HLA-A2.01-restricted epitope, GLCTLVAML, from the EBV-lytic cycle Ag BMLF1. TCR analysis of the highly amplified primary response to this epitope revealed markedly oligoclonal receptor usage among in vitro-derived clones, with similar clonotypes dominant in all three IM patients studied. Direct staining of IM T cell preparations with the A2.01/GLCTLVAML tetramer linked this oligoclonal epitope-specific response with appropriate Vbeta subset expansions in the patients' blood. These patients were studied again >2 years later, at which time TCR analysis of in vitro-reactivated clones suggested that rare clonotypes within the primary response had now come to dominate memory. Five additional A2. 01-positive IM patients were studied prospectively for Vbeta subset representation within primary and memory epitope specific populations as identified by tetramer staining. In each case, the primary response contained large Vbeta2, Vbeta16, or Vbeta22 components, and in three of five cases the originally dominant Vbeta was represented very poorly, if at all, in memory. We conclude 1) that an EBV epitope-specific primary response large enough to account for up to 10% CD8(+) T cells in IM blood may nevertheless be dominated by just a few highly expanded clonotypes, and 2) that with persistent viral challenge such dominant T cell clonotypes may be lost and replaced by others in memory. PMID- 11046005 TI - An early oxygen-dependent step is required for dexamethasone-induced apoptosis of immature mouse thymocytes. AB - The roles of oxygen and reactive oxygen intermediates in apoptosis are unclear at present. Although oxygen and reactive oxygen intermediates are not required for the execution of apoptosis, oxygen may be involved in at least some forms of apoptosis. In this study we show that dexamethasone (Dex)-induced apoptosis of immature mouse thymocytes is completely inhibited by hypoxic culture. In contrast, anti-CD95 thymocyte apoptosis is unaffected by hypoxia, indicating the existence of two forms of thymocyte apoptosis: an oxygen-dependent pathway (Dex induced) and an oxygen-independent pathway (anti-CD95 induced). Furthermore, hypoxia inhibited mitochondrial permeability transition (PT) in Dex-treated, but not in anti-CD95-treated, thymocytes, suggesting that the oxygen-sensitive step is upstream of mitochondria. Both Dex- and anti-CD95-induced PT and apoptosis were dependent on activation of IL-converting enzyme-like protease, as PT and apoptosis were inhibited by preincubation with Cbz-Val-Ala-Asp-fluoromethyl ketone, an irreversible inhibitor of IL-converting enzyme-like proteases. In addition, hypoxia inhibited the activation by Dex of caspase-3 (CPP32)-like proteases. Our data show that the private signaling pathways of Dex (oxygen dependent) and anti-CD95 (oxygen independent) both converge upstream of mitochondrial changes. The oxygen-dependent step in Dex-induced apoptosis lies upstream of caspase-3-like protease activation. Our observations support a model of apoptosis signaling in which independent pathways (oxygen dependent and oxygen independent) particular to each stimuli converge at a central point in the apoptotic cascade. PMID- 11046007 TI - Defining the requirements for peptide recognition in gene therapy-induced T cell tolerance. AB - Expression of a retrovirally transduced MHC class I Ag, H-2K(b) (K(b)), in bone marrow-derived cells leads to specific prolongation of K(b) disparate skin grafts. To examine the extent to which peptides derived from K(b) contribute to the induction of tolerance, retroviruses carrying mutant K(b) genes designed to enter separate pathways of Ag presentation were constructed. Thymectomized and CD8 T cell-depleted mice that had been irradiated and reconstituted with bone marrow cells expressing a secreted form of K(b) showed prolongation of K(b) disparate skin graft survival. Skin graft prolongation was not observed when similar experiments were performed using mice that were not CD8 T cell depleted. This suggests that hyporesponsiveness can be induced in CD4 T cells, but not CD8 T cells by Ags presented via the exogenous pathway of Ag processing. Modest prolongation of skin allografts was observed in mice reconstituted with bone marrow cells transduced with retroviruses carrying a gene encoding a mutant K(b) molecule expressed only in the cytoplasm. Prolongation was also observed in similar experiments in mice that were thymectomized and CD4 T cell depleted following complete reconstitution, but not in mice that were reconstituted and then thymectomized and CD8 T cell depleted. Thus, hyporesponsiveness can be induced in a subset of CD8 T cells by recognition of peptides derived from K(b) through both the direct and indirect pathways of Ag recognition, while CD4 T cell hyporesponsiveness to MHC class I disparate grafts occurs only through the indirect pathway of Ag recognition. PMID- 11046008 TI - T regulatory cells 1 inhibit a Th2-specific response in vivo. AB - We recently described a new population of CD4(+) regulatory T cells (Tr1) that inhibits proliferative responses of bystander T cells and prevents colitis induction in vivo through the secretion of IL-10. IL-10, which had been primarily described as a Th2-specific cytokine inhibiting Th1 responses, has displayed in several models a more general immune suppression on both types of effector T cell responses. Using an immediate hypersensitivity model in which BALB/c mice immunized with OVA (alum) normally generate Th2-dominated responses, we examined the ability of OVA-specific Tr1 T cell clones to inhibit OVA-specific cytokines and Ab responses. In contrast to Th2 or Th1 T cell clones, transfer of Tr1 T cell clones coincident with OVA immunization inhibited Ag-specific serum IgE responses, whereas IgG1 and IgG2a synthesis were not affected. This specific inhibition was mediated in part through IL-10 secretion as anti-IL-10 receptor Abs treatment reverted the inhibitory effect of Tr1 T cell clones. Although specifically targeted to IgE responses, Tr1 clones' inhibitory effects were more profound as they affected Ag-specific Th2 cell priming both in term of proliferative responses and cytokine secretion. These results suggest that regulatory T cells may play a fundamental role in maintaining the balance of the immune system to prevent allergic disorders. PMID- 11046009 TI - Receptor engagement on cells expressing a ligand for the tolerance-inducing molecule OX2 induces an immunoregulatory population that inhibits alloreactivity in vitro and in vivo. AB - Increased survival of C57BL/6 renal allografts following portal vein donor specific pretransplant immunization of C3H mice is associated with increased expression of the molecule OX2 seen on host dendritic cells, along with a marked polarization in cytokine production from lymphocytes harvested from the transplanted animals, with preferential production of IL-4, IL-10, and TGF-beta on donor-specific restimulation in vitro, and decreased production of IL-2, IFN gamma, and TNF-alpha compared with non-portal vein-immunized control transplanted mice. The increased renal allograft survival and the altered cytokine production are abolished by infusion of anti-mouse OX2 mAb (3B6). Infusion of a soluble OX2:Fc immunoadhesin can itself produce significant prolongation of xeno- and allografts in mice. We have used FITC-conjugated OX2:Fc to characterize cells expressing a ligand (OX2L) for OX2, and provide evidence that subpopulations of LPS-stimulated splenic macrophages, Con A-activated splenic T cells, and the majority (>80%) of gammadeltaTCR(+) T cells express this ligand. We show below that F4/80(+), OX2L(+) splenic macrophages, admixed with OX2:Fc, represent a potent immunosuppressive population capable of causing more profound inhibition of alloreactivity in vitro or in vivo than that seen using either OX2:Fc or OX2(+) (or OX2L(+)) cells alone. Immunoregulation by this OX2L(+) population occurs in an MHC-restricted fashion. PMID- 11046010 TI - The down-regulation of HLA-DM gene expression in rheumatoid arthritis is not related to their promoter polymorphism. AB - HLA-DM molecule, a class II-like heterodimer, is a critical factor of HLA class II-dependent Ag presentation. It acts as a molecular chaperone and also functions as a peptide editor favoring the presentation of high-stability peptides. Thus, it appears to skew the peptide repertoire presented to T cells. Variation in HLA DM expression has considerable effect on Ag presentation and regulation of these genes is likely to be a prerequisite to prevent autoimmunity. In this study, rheumatoid arthritis (RA) was chosen as a model of human autoimmune disease since its genetic susceptibility is known to be associated with the HLA-DR and -DM components. We described a limited nucleotide polymorphism in the HLA-DM promoters with functional impact on basal transcriptional activity and IFN-gamma induction as assessed in vitro. However, no difference of allele frequencies was found between controls and RA patients. Despite of this lack of association, expression of HLA-DM molecules was also investigated. Interestingly, an underexpression of HLA-DM transcripts and protein was shown in peripheral blood B cells from RA patients compared with controls or inflammatory arthritis patients. This underexpression does not affect HLA-DR genes and is responsible for a decrease of the DM:DR ratio in RA patients. This specific HLA-DM down-regulation is likely to have important consequences on Ag presentation and could participate in the autoimmune process in RA. PMID- 11046011 TI - Graded deletion and virus-induced activation of autoreactive CD4+ T cells. AB - We have examined factors governing the negative selection of autoreactive CD4(+) T cells in transgenic mice expressing low (HA12 mice) vs. high (HA104 mice) amounts of the influenza virus hemagglutinin (HA). When mated with TS1 mice that express a transgenic TCR specific for the I-Ed-restricted determinant site 1 (S1) of HA, thymocytes expressing high levels of the clonotypic TCR were deleted in both HA-transgenic lineages. However, through allelic inclusion, thymocytes with lower levels of the clonotypic TCR evaded deletion in TS1 x HA12 and TS1 x HA104 mice to graded degrees. Moreover, in both lineages, peripheral CD4(+) T cells could be activated by the S1 peptide in vitro, and by influenza virus in vivo. These findings indicate that allelic inclusion can allow autoreactive CD4(+) thymocytes to evade thymic deletion to varying extents reflecting variation in the expression of the self peptide, and can provide a basis for the activation of autoreactive peripheral T cells by viruses bearing homologues of self peptides ("molecular mimicry"). PMID- 11046012 TI - Functional differences between monocyte chemotactic protein-1 receptor A and monocyte chemotactic protein-1 receptor B expressed in a Jurkat T cell. AB - The monocyte chemotactic protein-1 (MCP-1) receptor (MCP-1R) is expressed on monocytes, a subpopulation of memory T lymphocytes, and basophils. Two alternatively spliced forms of MCP-1R, CCR2A and CCR2B, exist and differ only in their carboxyl-terminal tails. To determine whether CCR2A and CCR2B receptors function similarly, Jurkat T cells were stably transfected with plasmids encoding the human CCR2A or CCR2B gene. Nanomolar concentrations of MCP-1 induced chemotaxis in the CCR2B transfectants that express high, intermediate, and low levels of MCP-1R. Peak chemotactic activity was shifted to the right as receptor number decreased. Five-fold more MCP-1 was required to initiate chemotaxis of the CCR2A low transfectant, but the peak of chemotaxis was similar for the CCR2A and CCR2B transfectants expressing similar numbers of receptors. MCP-1-induced chemotaxis was sensitive to pertussis toxin, implying that both CCR2A and CCR2B are G(i)alpha protein coupled. MCP-1 induced a transient Ca(2+) flux in the CCR2B transfectant that was partially sensitive to pertussis toxin. In contrast, MCP-1 did not induce Ca(2+) flux in the CCR2A transfectant. Since MCP-1 can stimulate chemotaxis of the CCR2A transfectant without inducing Ca(2+) mobilization, Ca(2+) flux may not be required for MCP-1-induced chemotaxis in the Jurkat transfectants. These results indicate that functional differences exist between the CCR2A and CCR2B transfectants that can be attributed solely to differences in the carboxyl-terminal tail. PMID- 11046013 TI - The NFAT-related protein NFATL1 (TonEBP/NFAT5) is induced upon T cell activation in a calcineurin-dependent manner. AB - NFAT DNA binding complexes regulate programs of cellular activation and differentiation by translating receptor-dependent signaling events into specific transcriptional responses. NFAT proteins, originally defined as calcium/calcineurin-dependent regulators of cytokine gene transcription in T lymphocytes, are expressed in many different cell types and represent critical signaling intermediates that mediate an increasingly wide spectrum of biologic responses. Recent studies have identified a novel protein containing a region of similarity to the NFAT DNA binding domain. Here we demonstrate that this protein, designated NFATL1 (also known as tonicity enhancer binding protein and NFAT5) is expressed at high levels in the thymus but is undetectable in mature lymphocytes. However, NFATL1 can be induced in both primary quiescent T lymphocytes and differentiated Th1 and Th2 cell populations upon mitogen- or Ag receptor dependent activation. The induction of NFATL1 protein, as well as NFATL1 dependent transcription, is inhibited by cyclosporin A and FK506, and expression of constitutively active calcineurin induces NFATL1-dependent transcription. Overexpression of NFATc1 and inhibition of NFATc activity through the use of a dominant negative NFATc1 protein have no affect on NFATL1-dependent transcription, indicating that NFATc proteins do not play a role in the calcineurin-dependent induction of NFATL1. Interestingly, induction of NFATL1 by a hyperosmotic stimulus is not blocked by the inhibition of calcineurin. Moreover, osmotic stress response genes such as aldose reductase are not induced upon T cell activation. Thus inducible expression of NFATL1 represents a mechanism by which receptor-dependent signals as well as osmotic stress signals are translated into transcriptional responses that regulate cell function. PMID- 11046014 TI - Direct NK cell-mediated lysis of syngenic dorsal root ganglia neurons in vitro. AB - In contrast to extensive studies on the role of T and B lymphocytes in the pathogenesis of autoimmune diseases of the nervous system, little is known about NK cells and their potential role in the destruction of neural tissue. NK cells have been implicated in the selective death of sympathetic neurons resident in the superior cervical ganglia of rats after exposure to the drug guanethidine. This observation suggests that NK cells may function as principle effectors in immunological diseases of the nervous system. However, the direct mechanism of action of NK cells in this model is not known. In particular, it is not known whether NK cells can kill autologous neurons directly. The aim of the present study was to examine whether NK cells can kill directly dorsal root ganglia neurons cultured in vitro. We demonstrate that C57BL/6 (B6)-derived dorsal root ganglia neurons can be killed directly by syngenic IL-2-activated NK cells, and that this nerve cell lysis is dependent on the expression of perforin in the NK cells. NK cells were less effective in destroying neurons grown in the presence of glial cells. These observations indicate a potential role for NK cells in nerve cell degeneration in inflammatory diseases of the nervous system. PMID- 11046016 TI - Anatomical origin of dendritic cells determines their life span in peripheral lymph nodes. AB - Dendritic cells (DCs) exhibit considerable heterogeneity in their anatomical location, surface phenotype, and functional properties. In this study, we demonstrate that peripheral lymph nodes contain at least four major, functionally separable, and independently derived, DC subsets, which can be clearly demarcated by their CD11c, CD40, and CD8 expression pattern. Surprisingly, all DCs derived directly from the bone marrow, the myeloid- and the lymphoid-related subsets, turned over fast with t(1/2) of a couple of days. In contrast, DCs exported from the skin, both dermal and epidermal, accumulated 3- to 4-fold slower, turnover that is dramatically increased by cutaneous inflammation. PMID- 11046015 TI - Host T cells resist graft-versus-host disease mediated by donor leukocyte infusions. AB - Delayed lymphocyte infusions (DLIs) are used to treat relapse occurring post bone marrow transplantation (BMT) and to increase the donor chimerism in recipients receiving nonmyeloablative conditioning. As compared with donor lymphocytes given early post-BMT, DLIs are associated with a reduced risk of graft-vs-host disease (GVHD). The mechanism(s) responsible for such resistance have remained incompletely defined. We now have observed that host T cells present 3 wk after lethal total body irradiation, at the time of DLI, contribute to DLI-GVHD resistance. The infusion of donor splenocytes on day 0, a time when host bone marrow (BM)-derived T cells are absent, results in greater expansion than later post-BMT when host and donor BM-derived T cells coexist. Selective depletion of host T cells with anti-Thy1 allelic mAb increased the GVHD risk of DLI, indicating that a Thy1(+) host T cell regulated DLI-GVHD lethality. The conditions by which host T cells are required for optimal DLI resistance were determined. Recipients unable to express CD28 or 4-1BB were as susceptible to DLI GVHD as anti-Thy1 allelic mAb-treated recipients, indicating that CD28 and 4-1BB are critical to DLI-GVHD resistance. Recipients deficient in both perforin and Fas ligand but not individually were highly susceptible to DLI-GVHD. Recipients that cannot produce IFN-gamma were more susceptible to DLI-GVHD, whereas those deficient in IL-12 or p55 TNFRI were not. Collectively, these data indicate that host T cells, which are capable of generating antidonor CTL effector cells, are responsible for the impaired ability of DLI to induce GVHD. These same mechanisms may limit the efficacy of DLI in cancer therapy under some conditions. PMID- 11046017 TI - A subset of NKT cells that lacks the NK1.1 marker, expresses CD1d molecules, and autopresents the alpha-galactosylceramide antigen. AB - In the present report, we characterize a novel T cell subset that shares with the NKT cell lineage both CD1d-restriction and high reactivity in vivo and in vitro to the alpha-galactosylceramide (alpha-GalCer) glycolipid. These cells preferentially use the canonical Valpha14-Jalpha281 TCR-alpha-chain and Vbeta8 TCR-beta segments, and are stimulated by alpha-GalCer in a CD1d-dependent fashion. However, in contrast to classical NKT cells, they lack the NK1.1 marker and express high surface levels of CD1d molecules. In addition, this NK1.1(-) CD1d(high) T subset, further referred to as CD1d(high) NKT cells, can be distinguished by its unique functional features. Although NK1.1(+) NKT cells require exogenous CD1d-presenting cells to make them responsive to alpha-GalCer, CD1d(high) NKT cells can engage their own surface CD1d in an autocrine and/or paracrine manner. Furthermore, in response to alpha-GalCer, CD1d(high) NKT cells produce high amounts of IL-4 and moderate amounts of IFN-gamma, a cytokine profile more consistent with a Th2-like phenotype rather than the Th0-like phenotype typical of NK1.1(+) NKT cells. Our work reveals a far greater level of complexity within the NKT cell population than previously recognized and provides the first evidence for T cells that can be activated upon TCR ligation by CD1d restricted recognition of their ligand in the absence of conventional APCs. PMID- 11046018 TI - IFN-alpha suppresses activation of nuclear transcription factors NF-kappa B and activator protein 1 and potentiates TNF-induced apoptosis. AB - We and others have reported that IFN-alpha potentiates the apoptotic effects of TNF through a mechanism that is not understood. Because the nuclear transcription factors NF-kappaB and AP-1 have recently been reported to mediate anti-apoptosis and cell survival, we hypothesized that IFN-alpha potentiates the cytotoxic effects of TNF by suppressing TNF-induced activation of NF-kappaB and AP-1. We tested this hypothesis by pretreating human Jurkat T cells with IFN-alpha, which blocked TNF-induced activation of NF-kappaB and AP-1 in a time- and dose dependent manner as determined by EMSA. IFN-alpha blocked TNF-induced phosphorylation and degradation of the inhibitor subunit of NF-kappaB, and suppressed NF-kappaB and AP-1 activation induced by various other inflammatory stimuli. NF-kappaB-dependent reporter gene expression activated by TNF, TNFR1, TNF receptor-associated factor 2, and NF-kappaB-inducing kinase was also abrogated by IFN-alpha pretreatment. The suppression of NF-kappaB and AP-1 correlated with the potentiation of TNF-induced cytotoxicity and caspase activation. Overall our results suggest that IFN-alpha potentiates the apoptotic effects of TNF possibly by suppressing NF-kappaB and AP-1 activation. PMID- 11046019 TI - Potent costimulation of effector T lymphocytes by human collagen type I. AB - Purified, resting peripheral blood T lymphocytes were previously reported to undergo beta(1) integrin-dependent activation when cultured with anti-CD3 mAb coimmobilized with fibronectin, but not type I collagen. However, the extravascular T cells that encounter immobilized extracellular matrix proteins and are involved in disease pathogenesis have different properties from resting peripheral blood cells. In this study, we confirm that resting CD4(+) and CD8(+) T cells from peripheral blood are costimulated by immobilized fibronectin, but not type I collagen. In contrast, Ag- or mitogen-stimulated CD4(+) and CD8(+) T cell lines, used as models of the effector cells involved in disease, are more potently costimulated by type I collagen than fibronectin. The collagen-induced effects are similar in assays with serum-free medium and in more physiological assays in which anti-CD3 mAb is replaced by a threshold concentration of Ag and irradiated autologous PBMC as APC. The responses are beta(1) integrin dependent and mediated largely by very late Ag (VLA) 1 and 2, as shown by their up regulation on the T cell lines as compared with freshly purified resting PBL, and by the effects of blocking mAb. Reversed phase HPLC located the major costimulatory sequence(s) in the alpha1 chain of type I collagen, the structure of which was confirmed by amino acid sequencing. The results demonstrate the potential importance of type I collagen, an abundant extracellular matrix protein, in enhancing the activation of extravascular effector T cells in inflammatory disease, and point to a new immunotherapeutic target. PMID- 11046020 TI - Regulation of beta 1 integrin-mediated adhesion by T cell receptor signaling involves ZAP-70 but differs from signaling events that regulate transcriptional activity. AB - Stimulation of the CD3/TCR results within minutes in an increase in T cell adhesion mediated by beta(1) integrins. The biochemical pathways that control CD3 mediated increases in beta(1) integrin-mediated adhesion remain poorly characterized. In this study, the role of the tyrosine kinase ZAP-70 in the regulation of beta(1) integrin activity by the CD3/TCR was investigated. CD3 stimulation did not increase beta(1) integrin-mediated adhesion of the ZAP-70 deficient Jurkat T cell line, P116, to the beta(1) integrin ligand fibronectin. Reintroduction of wild-type ZAP-70, but not a kinase-inactive variant, K369R, corrected the adhesive defect observed in P116 T cells. In addition, the kinase inactive ZAP-70 mutant inhibited CD3-induced adhesion of primary human T cell blasts. Interestingly, a ZAP-70 mutant with a tyrosine to phenylalanine substitution at position 319 (Y319F) restored the adhesive defect in P116 T cells, even though Y319F ZAP-70 failed to fully reconstitute CD3-initiated NF-AT dependent transcription and tyrosine phosphorylation of the LAT adapter protein. Finally, expression of mutants of LAT and the SLP-76 adapter protein that modulate CD3-mediated activation of an NF-AT reporter gene failed to block CD3 induced increases in beta(1) integrin-mediated adhesion. These observations support a model in which the tyrosine kinase activity of ZAP-70 kinase is critical for regulation of beta(1) integrin activity by CD3/TCR. However, the signaling events downstream of ZAP-70 that regulate CD3/TCR-mediated activation of beta(1) integrin function exhibit key differences when compared with the signaling pathways that regulate transcriptional events initiated by CD3/TCR stimulation. PMID- 11046021 TI - IL-18 receptors, their role in ligand binding and function: anti-IL-1RAcPL antibody, a potent antagonist of IL-18. AB - IL-18 is critical in eliciting IFN-gamma production from Th1 cells both in vitro and in vivo. Th1 cells have been implicated in the pathogenesis of autoimmune disorders, making antagonists of IL-18 promising therapeutics. However, specificity and binding characteristics of IL-18R components have only been superficially explored. In this study, we show that IL-1R related protein 1 (IL 1Rrp1) and IL-1R accessory protein-like (IL-1RAcPL) confer responsiveness to IL 18 in a highly specific (no response to other IL-1 ligands) and unique manner (no functional pairing with other IL-1Rs and IL-1R-like molecules). Cotransfection with both receptor components resulted in expression of both low and high affinity binding sites for IL-18 (K:(d) of 11 and 0.4 nM, respectively). We prepared anti-IL-1RAcPL mAb TC30-28E3, which, in contrast to soluble R proteins, effectively inhibited the IL-18-induced activation of NF-kappaB. Quantitative PCR showed that Th1 but not Th2 cells are unique in that they coexpress IL-1Rrp1 and IL-1RAcPL. mAb TC30-28E3 inhibited IL-18-induced production of IFN-gamma by Th1 cells, being at least 10-fold more potent than anti-IL-18 ligand mAb. This study shows that IL-1RAcPL is highly specific to IL-18, is required for high affinity binding of IL-18, and that the anti-IL-1RAcPL mAb TC30-28E3 potently antagonizes IL-18 responses in vitro, providing a rationale for the use of anti-IL-1RAcPL Abs to inhibit Th1-mediated inflammatory pathologies. PMID- 11046022 TI - TGF-beta 1 and IFN-gamma direct macrophage activation by TNF-alpha to osteoclastic or cytocidal phenotype. AB - TNF-related activation-induced cytokine (TRANCE; also called receptor activator of NF-kappaB ligand (RANKL), osteoclast differentiation factor (ODF), osteoprotegerin ligand (OPGL), and TNFSF11) induces the differentiation of progenitors of the mononuclear phagocyte lineage into osteoclasts in the presence of M-CSF. Surprisingly, in view of its potent ability to induce inflammation and activate macrophage cytocidal function, TNF-alpha has also been found to induce osteoclast-like cells in vitro under similar conditions. This raises questions concerning both the nature of osteoclasts and the mechanism of lineage choice in mononuclear phagocytes. We found that, as with TRANCE, the macrophage deactivator TGF-beta(1) strongly promoted TNF-alpha-induced osteoclast-like cell formation from immature bone marrow macrophages. This was abolished by IFN-gamma. However, TRANCE did not share the ability of TNF-alpha to activate NO production or heighten respiratory burst potential by macrophages, or induce inflammation on s.c. injection into mice. This suggests that TGF-beta(1) promotes osteoclast formation not only by inhibiting cytocidal behavior, but also by actively directing TNF-alpha activation of precursors toward osteoclasts. The osteoclast appears to be an equivalent, alternative destiny for precursors to that of cytocidal macrophage, and may represent an activated variant of scavenger macrophage. PMID- 11046023 TI - Emergence of CD8+ T cells expressing NK cell receptors in influenza A virus infected mice. AB - Both innate and adaptive immune responses play an important role in the recovery of the host from viral infections. In the present report, a subset of cells coexpressing CD8 and NKR-P1C (NK1.1) was found in the lungs of mice infected with influenza A virus. These cells were detected at low numbers in the lungs of uninfected mice, but represented up to 10% of the total CD8(+) T cell population at day 10 postinfection. Almost all of the CD8(+)NK1.1(+) cells were CD8alphabeta(+)CD3(+)TCRalphabeta(+) and a proportion of these cells also expressed the NK cell-associated Ly49 receptors. Interestingly, up to 30% of these cells were virus-specific T cells as determined by MHC class I tetramer staining and by intracellular staining of IFN-gamma after viral peptide stimulation. Moreover, these cells were distinct from conventional NKT cells as they were also found at increased numbers in influenza-infected CD1(-/-) mice. These results demonstrate that a significant proportion of CD8(+) T cells acquire NK1.1 and other NK cell-associated molecules, and suggests that these receptors may possibly regulate CD8(+) T cell effector functions during viral infection. PMID- 11046024 TI - Stimulatory function of gp49A, a murine Ig-like receptor, in rat basophilic leukemia cells. AB - Murine gp49, a 49-kDa type I transmembrane glycoprotein, is a member of the Ig like receptors expressed on the surface of cells involved in natural immunity such as mast cells, NK cells, and macrophages. The two major subtypes, gp49A and gp49B, are encoded by two different genes adjacent to each other. gp49B contains an immunoreceptor tyrosine-based inhibitory motif in its cytoplasmic region and is known to function as an inhibitory molecule. In contrast, gp49A does not harbor any specific motif for signal transduction, nor has its physiological role been determined. Here we report on the stimulatory nature of gp49A by analyzing biochemical characteristics of chimeric molecules consisting of an ectodomain of Fc receptor and a C-terminal half of gp49A, namely the pretransmembrane, transmembrane, and cytoplasmic portions, expressed on the rat basophilic leukemia mast cell line. Cross-linking of the chimeric receptors evoked cytoplasmic calcium mobilization, PGD(2) release, and transcription of IL-3 and IL-4 genes, but did not elicit degranulation of the cells. The chimeric molecule could be expressed as a singlet and a homodimeric form on the cell surface. A pretransmembrane cysteine residue of gp49A was necessary for dimer formation. Dimerization was be necessary for their incorporation into glycolipid-enriched membrane fraction (GEM) upon cross-linking stimuli. The calcium mobilization response was inhibited by treatment of cells with methyl-beta-cyclodextrin, an inhibitor of GEM formation. Together with these results, it was strongly suggested that gp49A could be expressed as a homodimer and elicit activation signals that lead to calcium mobilization, eicosanoid production, and cytokine gene transcription through its incorporation into GEM. PMID- 11046025 TI - Ectopic human telomerase catalytic subunit expression maintains telomere length but is not sufficient for CD8+ T lymphocyte immortalization. AB - Like most somatic human cells, T lymphocytes have a limited replicative life span. This phenomenon, called senescence, presents a serious barrier to clinical applications that require large numbers of Ag-specific T cells such as adoptive transfer therapy. Ectopic expression of hTERT, the human catalytic subunit of the enzyme telomerase, permits fibroblasts and endothelial cells to avoid senescence and to become immortal. In an attempt to immortalize normal human CD8(+) T lymphocytes, we infected bulk cultures or clones of these cells with a retrovirus transducing an hTERT cDNA clone. More than 90% of transduced cells expressed the transgene, and the cell populations contained high levels of telomerase activity. Measuring the content of total telomere repeats in individual cells (by flowFISH) we found that ectopic hTERT expression reversed the gradual loss of telomeric DNA observed in control populations during long term culture. Telomere length in transduced cells reached the levels observed in freshly isolated normal CD8(+) lymphocytes. Nevertheless, all hTERT-transduced populations stopped to divide at the same time as nontransduced or vector-transduced control cells. When kept in IL-2 the arrested cells remained alive. Our results indicate that hTERT may be required but is not sufficient to immortalize human T lymphocytes. PMID- 11046026 TI - Differentiation of murine NK cells into distinct subsets based on variable expression of the IL-12R beta 2 subunit. AB - The cytokine IL-12 manifests its biological activity via interaction with a heterodimeric receptor (IL-12R) present on activated T and NK cells. The cDNAs for two IL-12R subunits have been cloned from human and mouse and designated IL 12Rbeta1 and IL-12Rbeta2. The expression of IL-12Rbeta2 on T cells is influenced by cytokines, particularly IL-4, IL-12, and IFN-gamma; however, little is known regarding regulation of IL-12R expression on NK cells. In this study we show that murine NK cells differentiate into IL-12Rbeta2(low) and IL-12Rbeta2(high) subsets after in vitro stimulation with IL-2 in the absence of exogenous polarizing cytokines. Subset development occurs gradually as NK cells expand in vitro and is generally complete by 8-12 days of culture. Once established, IL-12Rbeta2(low) and IL-12Rbeta2(high) subsets are highly stable in vitro and can be maintained for at least 20 days after FACS sorting. Formation of these NK subsets appears to be strain independent. Flow cytometric analyses demonstrate that both subsets express a number of NK-associated markers, including NK1.1, DX-5, Ly-49A, and Ly 49C, but that the Ly-49G2 class I inhibitory receptor is expressed predominantly on the IL-12Rbeta2(high) population. Both IL-12Rbeta2(low) and IL-12Rbeta2(high) NK cells respond to exogenous IL-12 by rapid production of high levels of IFN gamma and increased lytic activity against NK-sensitive YAC-1 target cells. Analyses of cytokine gene expression by RNase protection assay indicated that similar to the recently described human NK1 subset, both IL-12Rbeta2(high) and IL 12Rbeta2(low) murine NK subsets expressed high levels of IFN-gamma, whereas neither subset expressed mRNA for the NK2-associated cytokines IL-5 and IL-13. PMID- 11046027 TI - Dendritic cells prime in vivo alloreactive CD4 T lymphocytes toward type 2 cytokine- and TGF-beta-producing cells in the absence of CD8 T cell activation. AB - The mechanisms that influence the polarization of CD4 T cells specific for allogeneic MHC class II molecules in vivo are still poorly understood. We have examined the pathway of alloreactive CD4 T cell differentiation in a situation in which only CD4 T cells could be activated in vivo. In this report we show that priming of adult mice with allogeneic APC, in the absence of MHC class I-T cell interactions, induces a strong expansion of type 2 cytokine-producing allohelper T cells. These alloantigen-specific CD4 T cells directly recognize native allogeneic MHC class II molecules on APC and secrete, in addition to the prototypic Th2 cytokines IL-4, IL-5, and IL-10, large amounts of TGF-beta. The default Th2-phenotype acquisition is not genetically controlled and occurred both in BALB/c and C57BL/6 mice. CD8 T cells are the principal cell type that controls CD4 T cell differentiation in vivo. Furthermore, we demonstrate that strong Th2 priming can be induced not only with allogeneic splenocytes but also with a low number of bone marrow-derived dendritic cells. Finally, using a passive transfer system, we provide direct evidence that CD8 T cell expansion in situ promotes alloreactive Th1 cell development principally by preventing their default development to the Th2 pathway in a mechanism that is largely IFN-gamma independent. Therefore, this work demonstrates that type 2 cytokine production represents a dominant pathway of alloreactive CD4 T cell differentiation in adult mice, a phenomenon that was initially thought to occur only during the neonatal period. PMID- 11046028 TI - NF-kappa B is required for the positive selection of CD8+ thymocytes. AB - To examine the role of NF-kappaB in T cell development, we analyzed thymocyte ontogeny in transgenic (mutant I-kappaBalpha (mI-kappaBalpha)) mice that express a superinhibitory form of the NF-kappaB inhibitory protein, I-kappaBalpha (I kappaBalpha(A32/36)), under the control of the T cell-specific CD2 promoter and enhancer. Thymi from mI-kappaBalpha mice contained increased numbers of double positive (DP) and decreased numbers of both CD4(+) and CD8(+) single-positive cells, consistent with a block in DP thymocyte maturation. In addition, expression of CD69, a marker of positive selection, was decreased on DP thymocytes from the mI-kappaBalpha mice. To test directly whether NF-kappaB was required for positive or negative selection, we generated mI-kappaBalpha mice expressing the H-Y or 2C alphabeta TCR transgenes. Expression of the I kappaBalpha(A32/36) transgene caused a block in the positive selection of CD8(+) single-positive cells in both strains of TCR transgenic animals. In contrast, negative selection was unaffected by expression of the I-kappaBalpha(A32/36) transgene. Taken together, these results identified a NF-kappaB-dependent transcriptional pathway that is selectively required for the positive selection of CD8(+) thymocytes. PMID- 11046029 TI - Potent induction of alpha(1,3)-fucosyltransferase VII in activated CD4+ T cells by TGF-beta 1 through a p38 mitogen-activated protein kinase-dependent pathway. AB - Homing of effector T cells to sites of inflammation, particularly in the skin, is dependent on T cell expression of ligands for the endothelial selectins. Underlying expression of these ligands is the expression of alpha(1,3) fucosyltransferase VII (FucT-VII), a FucT essential for biosynthesis of selectin ligands. FucT-VII is sharply induced in activated T cells by IL-12, but cytokines other than IL-12 that induce FucT-VII and functional selectin ligands have not been identified, and are likely to be important in homing of T cells to other selectin-dependent sites. Screening of a number of cytokines known to be active on T cells identified only TGF-beta1 as able to up-regulate FucT-VII mRNA levels and selectin ligands on activated CD4 T cells. The sharp increase in FucT-VII induced by TGF-beta1 in activated T cells was completely blocked by pharmacologic inhibition of p38 mitogen-activated protein kinase, but was unaffected by mitogen activated protein/extracellular signal-related kinase kinase inhibitors. The selective ability of TGF-beta1 to induce selectin ligands on activated T cells is likely important for T cell homing to the gut, which is a strongly selectin dependent site, and correlates with the ability of TGF-beta1 to coordinately induce other gut-associated homing pathways. PMID- 11046030 TI - Two distinct stages in the transition from naive CD4 T cells to effectors, early antigen-dependent and late cytokine-driven expansion and differentiation. AB - Efficient peptide presentation by professional APC to naive and effector CD4 T cells in vitro is limited to the first 1-2 days of culture, but is nonetheless optimum for effector expansion and cytokine production. In fact, prolonging Ag presentation leads to high levels of T cell death, decreased effector expansion, and decreased cytokine production by recovered effectors. Despite the absence of Ag presentation beyond day 2, T cell division continues at a constant rate throughout the 4-day culture. The Ag-independent later stage depends on the presence of IL-2, and we conclude optimum effector generation depends on an initial 2 days of TCR stimulation followed by an additional 2 days of Ag independent, cytokine driven T cell expansion and differentiation. PMID- 11046031 TI - Microbial and T cell-derived stimuli regulate antigen presentation by dendritic cells in vivo. AB - B cells and dendritic cells (DC) internalize and degrade exogenous Ags and present them as peptides bound to MHC class II molecules for scrutiny by CD4(+) T cells. Here we use an Ab specific for a processed form of the model Ag, hen egg lysozyme (HEL), to demonstrate that this protein is not efficiently presented by lymph node DC following s.c. immunization. HEL presentation by the DC can be dramatically enhanced upon coinjection of a microbial adjuvant, which appears to act by enhancing peptide loading onto MHC class II. CD40 cross-linking or the presence of a high frequency of T cells specific for HEL can similarly improve presentation by DC in vivo. For any of these activating stimuli, CD8alpha(+) DC consistently display the highest proportion of HEL-loaded MHC class II molecules. These data indicate that exogenous Ags can be displayed to T cells in lymphoid tissues by a large cohort of resident DC whose presentation is regulated by innate and adaptive stimuli. Our data further reveal the existence of a feedback mechanism that augments Ag presentation during cognate APC-T cell interactions. PMID- 11046032 TI - Mouse inducible costimulatory molecule (ICOS) expression is enhanced by CD28 costimulation and regulates differentiation of CD4+ T cells. AB - The inducible costimulatory (ICOS) molecule is expressed by activated T cells and has homology to CD28 and CD152. ICOS binds B7h, a molecule expressed by APC with homology to CD80 and CD86. To investigate regulation of ICOS expression and its role in Th responses we developed anti-mouse ICOS mAbs and ICOS-Ig fusion protein. Little ICOS is expressed by freshly isolated mouse T cells, but ICOS is rapidly up-regulated on most CD4(+) and CD8(+) T cells following stimulation of the TCR. Strikingly, ICOS up-regulation is significantly reduced in the absence of CD80 and CD86 and can be restored by CD28 stimulation, suggesting that CD28 CD80/CD86 interactions may optimize ICOS expression. Interestingly, TCR transgenic T cells differentiated into Th2 expressed significantly more ICOS than cells differentiated into Th1. We used two methods to investigate the role of ICOS in activation of CD4(+) T cells. First, CD4(+) cells were stimulated with beads coated with anti-CD3 and either B7h-Ig fusion protein or control Ig fusion protein. ICOS stimulation enhanced proliferation of CD4(+) cells and production of IFN-gamma, IL-4, and IL-10, but not IL-2. Second, TCR-transgenic CD4(+) T cells were stimulated with peptide and APC in the presence of ICOS-Ig or control Ig. When the ICOS:B7h interaction was blocked by ICOS-Ig, CD4(+) T cells produced more IFN-gamma and less IL-4 and IL-10 than CD4(+) cells differentiated with control Ig. These results demonstrate that ICOS stimulation is important in T cell activation and that ICOS may have a particularly important role in development of Th2 cells. PMID- 11046033 TI - Blockade of costimulation through B7/CD28 inhibits experimental autoimmune uveoretinitis, but does not induce long-term tolerance. AB - It has been reported that costimulation blockade can result in T cell anergy. We investigated the effects of blocking costimulatory molecules in vivo on the development of experimental autoimmune uveoretinitis (EAU), a model for autoimmune uveitis in humans that is induced in mice by immunization with the retinal Ag interphotoreceptor retinoid binding protein. B10.A mice immunized with a uveitogenic regimen of interphotoreceptor retinoid-binding protein were treated with Abs to B7.1 and B7.2 for 2 wk. Evaluation of EAU and immunological responses 1 wk later showed that disease had been abrogated, and cellular responses were suppressed. To determine whether the costimulation blockade resulted in tolerance, adult-thymectomized mice immunized for uveitis and treated with anti B7 or anti-CD28 were rechallenged for uveitis induction 5 wk after the initial immunization. Although confirmed to be disease free after the initial immunization, both anti-B7- and anti-CD28-treated mice developed severe EAU and elevated cellular responses after the rechallenge, equivalent to those of control mice. We conclude that in this model costimulatory blockade in vivo prevents the development of autoimmune disease, but does not result in long-term tolerance. The data are compatible with the interpretation that B7/CD28 blockade prevents generation of effector, but not of memory, T cells. PMID- 11046034 TI - Functional defects of NK cells treated with chloroquine mimic the lytic defects observed in perforin-deficient mice. AB - NK cells are the primary effectors mediating acute rejection of incompatible bone marrow cell grafts. To reduce rejection, we evaluated the ability of chloroquine (CHQ) to prevent perforin-dependent NK cell activity. Perforin is a key cytotoxic component released from the lytic granules of activated NK cells. Generation of functional perforin requires an acidic protease activity that occurs in the secretory, lytic lysosomes. Our hypothesis was that CHQ, a lysosomotropic reagent, would raise the pH of the acidic compartment in which perforin is processed and thereby block perforin maturation and cytotoxicity. We have measured NK cytotoxicity in vivo by clearance of YAC-1 tumor cells from the lungs and by rejection of incompatible bone marrow transplants and in vitro by cytolysis of YAC-1 and Jurkat cells. The engraftment of bone marrow cells was monitored by recolonization of the spleen with hemopoietic cells from transplants of MHC class I-deficient bone marrow cells into lethally irradiated recipient mice. Transplant rejection was compared in two inbred strains of mice: 129, which apparently use perforin-dependent cytotoxicity, and C57BL/6, in which rejection can be perforin-independent. CHQ treatment reduced NK cell activity in 129 mice in which perforin is important for mediating rejection. CHQ affected the fraction of NK cell cytolysis that was Fas independent. In addition, we found that CHQ prevents perforin processing by LAK cells in vitro. These data indicate that CHQ may impair rejection of incompatible bone marrow transplants and other functions mediated by NK and cytotoxic T cells. PMID- 11046035 TI - Allergen-induced airway hyperreactivity is diminished in CD81-deficient mice. AB - We demonstrated previously that CD81(-/-) mice have an impaired Th2 response. To determine whether this impairment affected allergen-induced airway hyperreactivity (AHR), CD81(-/-) BALB/c mice and CD81(+/+) littermates were sensitized i.p. and challenged intranasally with OVA. Although wild type developed severe AHR, CD81(-/-) mice showed normal airway reactivity and reduced airway inflammation. Nevertheless, OVA-specific T cell proliferation was similar in both groups of mice. Analysis of cytokines secreted by the responding CD81(-/ ) T cells, particularly those derived from peribronchial draining lymph nodes, revealed a dramatic reduction in IL-4, IL-5, and IL-13 synthesis. The decrease in cytokine production was not due to an intrinsic T cell deficiency because naive CD81(-/-) T cells responded to polyclonal Th1 and Th2 stimulation with normal proliferation and cytokine production. Moreover, there was an increase in T cells and a decrease in B cells in peribronchial lymph nodes and in spleens of immunized CD81(-/-) mice compared with wild-type animals. Interestingly, OVA specific Ig levels, including IgE, were similar in CD81(-/-) and CD81(+/+) mice. Thus, CD81 plays a role in the development of AHR not by influencing Ag-specific IgE production but by regulating local cytokine production. PMID- 11046036 TI - Regulation of cell surface expression of CTLA-4 by secretion of CTLA-4-containing lysosomes upon activation of CD4+ T cells. AB - CTLA-4 is expressed on the surface of activated T cells and negatively regulates T cell activation. Because a low-level expression of CTLA-4 on the cell surface is sufficient to induce negative signals in T cells, the surface expression of CTLA-4 is strictly regulated. We previously demonstrated that the association of CTLA-4 with the clathrin-associated adaptor complex AP-2 induces internalization of CTLA-4 and keeps the surface expression low. However, the mechanism to induce high expression on the cell surface upon stimulation has not yet been clarified. To address this, we investigated the intracellular dynamics of CTLA-4 by analyzing its localization and trafficking in wild-type and mutant CTLA-4 transfected Th1 clones. CTLA-4 is accumulated in intracellular granules, which we identified as lysosomes. CTLA-4 is degraded in lysosomes in a short period, and the degradation process may serve as one of the mechanisms to regulate CTLA-4 expression. Upon TCR stimulation, CTLA-4-containing lysosomes are secreted as proven by the secretion of cathepsin D and beta-hexosaminidase in parallel with the increase of surface expression of CTLA-4 and lysosomal glycoprotein 85, a lysosomal marker. These results suggest that the cell surface expression of CTLA 4 is up-regulated upon stimulation by utilizing a mechanism of secretory lysosomes in CD4(+)T cells. PMID- 11046037 TI - The role of thymus-expressed chemokine and its receptor CCR9 on lymphocytes in the regional specialization of the mucosal immune system. AB - Chemokines play an important role in the migration of leukocytes at sites of inflammation, and some constitutively expressed chemokines may direct lymphocyte trafficking within lymphoid organs and peripheral tissues. Thymus-expressed chemokine (TECK or Ckbeta-15/CCL25), which signals through the chemokine receptor CCR9, is constitutively expressed in the thymus and small intestine but not colon, and chemoattracts a small fraction of PBLs that coexpress the integrin alpha(4)beta(7). Here we show that TECK is expressed in the human small bowel but not colon by endothelial cells and a subset of cells in intestinal crypts and lamina propria. CCR9 is expressed in the majority of freshly isolated small bowel lamina propria mononuclear cells (LPMC) and at significantly higher levels compared with colonic LPMC or PBL. TECK was selectively chemotactic for small bowel but not colonic LPMC in vitro. The TECK-induced chemotaxis was sensitive to pertussis toxin and partially inhibited by Abs to CCR9. TECK attracts predominantly the T cell fraction of small bowel LPMC, whereas sorted CD3(+)CCR9(+) and CD3(+)CCR9(-) lymphocytes produce similar Th1 or Th2 cytokines at the single cell level. Collectively, our data suggest that the selective expression of TECK in the small bowel underlie the homing of CCR9(+) intestinal memory T cells to the small bowel rather than to the colon. This regional specialization implies a segregation of small intestinal from colonic immune responses. PMID- 11046038 TI - Vaginal mucosa serves as an inductive site for tolerance. AB - These data demonstrate that tolerance can be induced by vaginal Ag exposure. In these experiments, mice were given vaginal agarose gel suppositories containing either 5 mg OVA or saline for 6 h. Mice were given suppositories either during the estrous (estrogen dominant) or diestrous (progesterone dominant) stage of the estrous cycle. Mice were restrained during the inoculation period to prevent orovaginal transmission of the Ag. After 1 wk, mice were immunized s. c. with OVA in CFA. After 3 wk, mice were tested for delayed-type hypersensitivity responses by measuring footpad swelling and measuring in vitro proliferation of lymphocytes to Ag. Using ELISA, the magnitude of the serum Ab response was also measured. In some mice, FITC conjugated to OVA was used to track the dissemination of the protein into the systemic tissues. The magnitude of footpad swelling was significantly reduced in mice receiving OVA-containing suppositories during estrus compared with mice receiving saline suppositories. Concomitant decreases in the Ag-specific proliferative response were also observed in lymph node lymphocytes and splenocytes. Conversely, mice inoculated during diestrus did not show a decreased response to Ag by either footpad response or in vitro proliferation. Serum Ab titers in the estrus-inoculated mice did not decrease significantly. These data demonstrate that the reproductive tract can be an inductive site for mucosally induced tolerance. However, unlike other mucosal sites such as the lung and gastrointestinal tract, reproductive tract tolerance induction is hormonally regulated. PMID- 11046039 TI - TNFR-associated factor family protein expression in normal tissues and lymphoid malignancies. AB - TNFR-associated factors (TRAFs) constitute a family of adapter proteins that associate with particular TNF family receptors. Humans and mice contain six TRAF genes, but little is known about their in vivo expression at the single cell level. The in vivo locations of TRAF1, TRAF2, TRAF5, and TRAF6 were determined in human and mouse tissues by immunohistochemistry. Striking diversity was observed in the patterns of immunostaining obtained for each TRAF family protein, suggesting their expression is independently regulated in a cell type-specific manner. Dynamic regulation of TRAFs was observed in cultured PBLs, where anti-CD3 Abs, mitogenic lectins, and ILs induced marked increases in the steady-state levels of TRAF1, TRAF2, TRAF5, and TRAF6. TRAF1 was also highly inducible by CD40 ligand in cultured germinal center B cells, whereas TRAF2, TRAF3, TRAF5, and TRAF6 were relatively unchanged. Analysis of 83 established human tumor cell lines by semiquantitative immunoblotting methods revealed tendencies of certain cancer types to express particular TRAFs. For example, expression of TRAF1 was highly restricted, with B cell lymphomas consistently expressing this TRAF family member. Consistent with results from tumor cell lines, immunohistochemical analysis of 232 non-Hodgkin lymphomas revealed TRAF1 overexpression in 112 (48%) cases. TRAF1 protein levels were also elevated in circulating B cell chronic lymphocytic leukemia specimens (n = 49) compared with normal peripheral blood B cells (p = 0.01), as determined by immunoblotting. These findings contribute to an improved understanding of the cell-specific roles of TRAFs in normal tissues and provide evidence of altered TRAF1 expression in lymphoid malignancies. PMID- 11046040 TI - Functional uncoupling of the Janus kinase 3-Stat5 pathway in malignant growth of human T cell leukemia virus type 1-transformed human T cells. AB - Human T cell leukemia virus type 1 (HTLV-1) transforms cytokine-dependent T lymphocytes and causes adult T cell leukemia. Janus tyrosine kinase (Jak)3 and transcription factors Stat5a and Stat5b are essential for the proliferation of normal T cells and are constitutively hyperactivated in both HTLV-1-transformed human T cell lines and lymphocytes isolated from HTLV-1-infected patients; therefore, a critical role for the Jak3-Stat5 pathway in the progression of this disease has been postulated. We recently reported that tyrphostin AG-490 selectively blocked IL-2 activation of Jak3/Stat5 and growth of murine T cell lines. Here we demonstrate that disruption of Jak3/Stat5a/b signaling with AG-490 (50 microM) blocked the proliferation of primary human T lymphocytes, but paradoxically failed to inhibit the proliferation of HTLV-1-transformed human T cell lines, HuT-102 and MT-2. Structural homologues of AG-490 also inhibited the proliferation of primary human T cells, but not HTLV-1-infected cells. Disruption of constitutive Jak3/Stat5 activation by AG-490 was demonstrated by inhibition of 1) tyrosine phosphorylation of Jak3, Stat5a (Tyr(694)), and Stat5b (Tyr(699)); 2) serine phosphorylation of Stat5a (Ser(726)) as determined by a novel phosphospecific Ab; and 3) Stat5a/b DNA binding to the Stat5-responsive beta casein promoter. In contrast, AG-490 had no effect on DNA binding by p50/p65 components of NF-kappaB, a transcription factor activated by the HTLV-1-encoded phosphoprotein, Tax. Collectively, these data suggest that the Jak3-Stat5 pathway in HTLV-1-transformed T cells has become functionally redundant for proliferation. Reversal of this functional uncoupling may be required before Jak3/Stat5 inhibitors will be useful in the treatment of this malignancy. PMID- 11046041 TI - CD30 signals integrate expression of cytotoxic effector molecules, lymphocyte trafficking signals, and signals for proliferation and apoptosis. AB - Although CD30 has long been recognized as an important marker on many lymphomas of diverse origin and as activation molecule on B cells and T cells, its primary function has remained obscure. We now report that CD30 signals may serve to inhibit effector cell activity by integrating gene expression changes of several pathways important for cytotoxic NK and T cell effector function. In the large granular lymphoma line YT, CD30 signals down-regulate the expression of cytotoxic effector molecules, Fas ligand, perforin, granzyme B, and abrogate cytotoxicity. c-myc, a regulator of proliferation and an upstream regulator of Fas ligand expression, is completely suppressed by CD30. Furthermore, CD30 signals strongly induce CCR7, suggesting a role for CD30 signals in the homing of lymphocytes to lymph nodes. The up-regulation of Fas, death receptor 3, and TNF-related apoptosis-inducing ligand by CD30 indicates an increase in susceptibility to apoptotic signals whereas up-regulation of TNFR-associated factor 1 and cellular inhibitor of apoptosis 2 protect cells from certain types of apoptosis. Using gene microarrays, 750 gene products were induced and 90 gene products were suppressed >2-fold by CD30 signals. Signals emanating from CD30 use both TNFR associated factor 2-dependent and -independent pathways. The integration of CD30 signals in a lymphoma line suggests that CD30 can down-modulate lymphocyte effector function and proliferation while directing the cells to lymph nodes and increasing their susceptibility to certain apoptotic signals. These studies may provide a molecular mechanism for the recently observed CD30-mediated suppression of CTL activity in vivo in a diabetes model. PMID- 11046042 TI - Recombinant anti-human HER2/neu IgG3-(GM-CSF) fusion protein retains antigen specificity and cytokine function and demonstrates antitumor activity. AB - Anti-HER2/neu therapy of human HER2/neu-expressing malignancies such as breast cancer has shown only partial success in clinical trials. To expand the clinical potential of this approach, we have genetically engineered an anti-HER2/neu IgG3 fusion protein containing GM-CSF. Anti-HER2/neu IgG3-(GM-CSF) expressed in myeloma cells was correctly assembled and secreted. It was able to target HER2/neu-expressing cells and to support growth of a GM-CSF-dependent murine myeloid cell line, FDC-P1. The Ab fusion protein activated J774.2 macrophage cells so that they exhibit an enhanced cytotoxic activity and was comparable to the parental Ab in its ability to effect Ab-dependent cellular cytotoxicity mediated tumor cell lysis. Pharmacokinetic studies showed that anti-HER2/neu IgG3 (GM-CSF) is stable in the blood. Interestingly, the half-life of anti-HER2/neu IgG3-(GM-CSF) depended on the injected dose with longer in vivo persistence observed at higher doses. Biodistribution studies showed that anti-HER2/neu IgG3 (GM-CSF) is mainly localized in the spleen. In addition, anti-HER2/neu IgG3-(GM CSF) was able to target the HER2/neu-expressing murine tumor CT26-HER2/neu and enhance the immune response against the targeted Ag HER2/neu. Anti-HER2/neu IgG3 (GM-CSF) is able to enhance both Th1- and Th2-mediated immune responses and treatment with this Ab fusion protein resulted in significant retardation in the growth of s.c. CT26-HER2/neu tumors. Our results suggest that anti-HER2/neu IgG3 (GM-CSF) fusion protein is useful in the treatment of HER2/neu-expressing tumors. PMID- 11046043 TI - The inference of antigen selection on Ig genes. AB - Analysis of somatic mutations in V regions of Ig genes is important for understanding various biological processes. It is customary to estimate Ag selection on Ig genes by assessment of replacement (R) as opposed to silent (S) mutations in the complementary-determining regions and S as opposed to R mutations in the framework regions. In the past such an evaluation was performed using a binomial distribution model equation, which is inappropriate for Ig genes in which mutations have four different distribution possibilities (R and S mutations in the complementary-determining region and/or framework regions of the gene). In the present work, we propose a multinomial distribution model for assessment of Ag selection. Side-by-side application of multinomial and binomial models on 86 previously established Ig sequences disclosed 8 discrepancies, leading to opposite statistical conclusions about Ag selection. We suggest the use of the multinomial model for all future analysis of Ag selection. PMID- 11046044 TI - Receptor for activated C-kinase (RACK-1), a WD motif-containing protein, specifically associates with the human type I IFN receptor. AB - The cytoplasmic domain of the human type I IFN receptor chain 2 (IFNAR2c or IFN alphaRbetaL) was used as bait in a yeast two-hybrid system to identify novel proteins interacting with this region of the receptor. We report here a specific interaction between the cytoplasmic domain of IFN-alphaRbetaL and a previously identified protein, RACK-1 (receptor for activated C kinase). Using GST fusion proteins encoding different regions of the cytoplasmic domain of IFN-alphaRbetaL, the minimum site for RACK-1 binding was mapped to aa 300-346. RACK-1 binding to IFN-alphaRbetaL did not require the first 91 aa of RACK-1, which includes two WD domains, WD1 and WD2. The interaction between RACK-1 and IFN-alphaRbetaL, but not the human IFN receptor chain 1 (IFNAR1 or IFN-alphaRalpha), was also detected in human Daudi cells by coimmunoprecipitation. RACK-1 was shown to be constitutively associated with IFN-alphaRbetaL, and this association was not effected by stimulation of Daudi cells with type I IFNs (IFN-beta1b). RACK-1 itself did not become tyrosine phosphorylated upon stimulation of Daudi cells with IFN-beta1b. However, stimulation of cells with either IFN-beta1b or PMA did result in an increase in detectable immunofluorescence and intracellular redistribution of RACK-1. PMID- 11046045 TI - DNA vaccination against rat her-2/Neu p185 more effectively inhibits carcinogenesis than transplantable carcinomas in transgenic BALB/c mice. AB - The ability of vaccination with plasmids coding for the extracellular and the transmembrane domain of the product of transforming rat Her-2/neu oncogene (r p185) to protect against r-p185(+) transplantable carcinoma (TUBO) cells and mammary carcinogenesis was evaluated. In normal BALB/c mice, DNA vaccination elicits anti-r-p185 Ab, but only a marginal CTL reactivity, and protects against a TUBO cell challenge. Massive reactive infiltration is associated with TUBO cell rejection. In BALB/c mice transgenic for the rat Her-2/neu gene (BALB-neuT), DNA vaccination elicits a lower anti-r-p185 Ab response, no CTL activity and only incompletely protects against TUBO cells, but markedly hampers the progression of carcinogenesis. At 33 wk of age, when control BALB-neuT mice display palpable tumors in all mammary glands, about 60% of immunized mice are tumor free, and tumor multiplicity is markedly reduced. Tumor-free mammary glands still display the atypical hyperplasia of the early stages of carcinogenesis, and a marked down modulation of r-p185, along with a massive reactive infiltrate. However, BALB neuT mice protected against mammary carcinogenesis fail to efficiently reject a TUBO cell challenge. This suggests that the mechanisms required for the rejection of transplantable tumors may not coincide with those that inhibit the slow progression of carcinogenesis. PMID- 11046046 TI - Functional modulation of human macrophages through CD46 (measles virus receptor): production of IL-12 p40 and nitric oxide in association with recruitment of protein-tyrosine phosphatase SHP-1 to CD46. AB - Human CD46, formerly membrane cofactor protein, binds and inactivates complement C3b and serves as a receptor for measles virus (MV), thereby protecting cells from homologous complement and sustaining systemic measles infection. Suppression of cell-mediated immunity, including down-regulation of IL-12 production, has been reported on macrophages (Mphi) by cross-linking their CD46. The intracellular events responsible for these immune responses, however, remain unknown. In this study, we found that 6- to 8-day GM-CSF-treated peripheral blood monocytes acquired the capacity to recruit protein-tyrosine phosphatase SHP-1 to their CD46 and concomitantly were able to produce IL-12 p40 and NO. These responses were induced by stimulation with mAbs F(ab')(2) against CD46 that block MV binding or by a wild-type MV strain Kohno MV strain (KO; UV treated or untreated) that was reported to induce early phase CD46 down-regulation. Direct ligation of CD46 by these reagents, but not intracellular MV replication, was required for these cellular responses. Interestingly, the KO strain failed to replicate in the 6- to 8-day GM-CSF-cultured Mphi, while other MV strains replicated to form syncytia under the same conditions. When stimulated with the KO strain, rapid and transient dissociation of SHP-1 from CD46 was observed. These and previous results provide strong evidence that CD46 serves as a signal modulatory molecule and that the properties of ligands determine suppression or activation of an innate immune system at a specific maturation stage of human Mphi. PMID- 11046047 TI - Enhanced murine macrophage TNF receptor shedding by cytosine-guanine sequences in oligodeoxynucleotides. AB - The immunomodulatory role of unmethylated cytosine-guanine sequences (CpG) in bacterial DNA has been well documented. We have previously demonstrated that murine macrophage-like RAW 264.7 cells respond to CpG DNA with an increase in the proinflammatory cytokine, TNF-alpha, in both a dose-dependent and time-dependent manner. In addition, CpG DNA stimulates a significant, though delayed, secretion of the anti-inflammatory cytokine IL-10. Because TNF-alpha and TNFR (TNFRI and II) expression are tightly regulated responses, we hypothesized that CpG containing oligodeoxynucleotide (CpG ODN) would also affect TNFRI and II shedding. Using both murine peritoneal macrophages and RAW 264.7 cells, we demonstrated a significant, time-dependent increase in soluble TNFRI and TNFRII production with CpG ODN stimulation. RAW 264.7 cells treated with CpG ODN had a transient increase in membrane TNFRII expression, but not TNFRI. Both types of TNFR mRNA were also up-regulated by CpG ODN, and addition of the transcriptional inhibitor actinomycin D abrogated the effect of CpG ODN on TNFR mRNA and protein expression. Addition of anti-IL-10 and anti-TNF-alpha Abs did not change these results. The addition of plate-bound anti-TNF receptor Abs to this system increased the amount of bioactive TNF, implying that these receptors are acting as inhibitors of TNF activity. These results suggest that the de novo, non-IL-10- and non-TNF-alpha-dependent transcription, translation, and shedding of TNFRs are additional potential counterinflammatory effects of CpG DNA. PMID- 11046048 TI - The serpin secreted by Brugia malayi microfilariae, Bm-SPN-2, elicits strong, but short-lived, immune responses in mice and humans. AB - Understanding the basic immunology of an infectious disease requires insight into the pattern of T cell reactivity and specificity. Although lymphatic filariasis is a major tropical disease, the predominant T cell Ags of filarial species such as Brugia malayi are still undefined. We have now identified a prominent T cell Ag from B. malayi microfilariae (Mf) as Bm-SPN-2, a serpin secreted exclusively by this stage. Mf-infected mice mounted strong, but short-lived, Bm-SPN-2 specific Th1 responses, measured by in vitro production of IFN-gamma, but not IL 4 or IL-5, 14 days postinfection. By day 35, responsiveness to Bm-SPN-2 was lost despite enhanced reactivity to whole Mf extract. Single immunization with Mf extract also stimulated typical Th1 reactions to Bm-SPN-2, but IgG1 Ab responses dominated after repeated immunizations. Human patients displayed potent humoral responses to Bm-SPN-2 in both IgG1 and IgG4 subclasses. Thus, 100% (20 of 20) of the microfilaremic (MF(+)) patients bore IgG4 responses to Bm-SPN-2, while only 30% of endemic normal subjects were similarly positive. Following chemotherapy, Bm-SPN-2-specific Abs disappeared in 12 of 13 MF(+) patients, although the majority remained seropositive for whole parasite extract. PBMC from most, but not all, endemic subjects were induced to secrete IFN-gamma when stimulated with Bm-SPN-2. These findings demonstrate that Bm-SPN-2 is recognized by both murine and human T and B cells and indicate that their responses are under relatively stringent temporal control. This study also provides the first example of a stage specific secreted molecule that acts as a major T cell Ag from filarial parasites and is a prime candidate for a serodiagnostic probe. PMID- 11046049 TI - Mucosal and plasma IgA from HIV-1-exposed uninfected individuals inhibit HIV-1 transcytosis across human epithelial cells. AB - HIV-1-specific IgA has been described in the genital tract and plasma of HIV-1 highly exposed, persistently seronegative (HEPS) individuals, and IgA from these sites has been shown to neutralize HIV-1. This study examines the ability of IgA isolated from HEPS individuals to inhibit transcytosis across a tight epithelial cell layer. A Transwell system was established to model HIV-1 infection across the human mucosal epithelium. The apical-basolateral transcytosis of primary HIV 1 isolates across this mucosal model was examined in the presence and the absence of IgA isolated from the genital tract, saliva, and plasma of HEPS individuals enrolled in both a sex worker cohort in Nairobi, Kenya, and a discordant couple cohort in Italy. In the absence of IgA, HIV-1 primary isolates were actively transported across the epithelial membrane and were released on the opposite side of the barrier. These transcytosed HIV-1 particles retained their ability to infect human mononuclear cells. However, IgA purified from the mucosa and plasma of HEPS individuals was able to inhibit HIV-1 transcytosis. Inhibition was seen in three of six cervicovaginal fluid samples, five of 10 saliva samples, and three of six plasma samples against at least one of the two primary HIV-1 isolates tested. IgA from low risk, healthy control subjects had no inhibitory effect on HIV-1 transcytosis. The ability of mucosal and plasma IgA to inhibit HIV-1 transcytosis across the mucosal epithelium may represent an important mechanism for protection against the sexual acquisition of HIV-1 infection in HEPS individuals. PMID- 11046050 TI - Archaeosomes induce long-term CD8+ cytotoxic T cell response to entrapped soluble protein by the exogenous cytosolic pathway, in the absence of CD4+ T cell help. AB - The unique ether glycerolipids of Archaea can be formulated into vesicles (archaeosomes) with strong adjuvant activity for MHC class II presentation. Herein, we assess the ability of archaeosomes to facilitate MHC class I presentation of entrapped protein Ag. Immunization of mice with OVA entrapped in archaeosomes resulted in a potent Ag-specific CD8(+) T cell response, as measured by IFN-gamma production and cytolytic activity toward the immunodominant CTL epitope OVA(257-264). In contrast, administration of OVA with aluminum hydroxide or entrapped in conventional ester-phospholipid liposomes failed to evoke significant CTL response. The archaeosome-mediated CD8(+) T cell response was primarily perforin dependent because CTL activity was undetectable in perforin deficient mice. Interestingly, a long-term CTL response was generated with a low Ag dose even in CD4(+) T cell deficient mice, indicating that the archaeosomes could mediate a potent T helper cell-independent CD8(+) T cell response. Macrophages incubated in vitro with OVA archaeosomes strongly stimulated cytokine production by OVA-specific CD8(+) T cells, indicating that archaeosomes efficiently delivered entrapped protein for MHC class I presentation. This processing of Ag was Brefeldin A sensitive, suggesting that the peptides were transported through the endoplasmic reticulum and presented by the cytosolic MHC class I pathway. Finally, archaeosomes induced a potent memory CTL response to OVA even 154 days after immunization. This correlated to strong Ag-specific up regulation of CD44 on splenic CD8(+) T cells. Thus, delivery of proteins in self adjuvanting archaeosomes represents a novel strategy for targeting exogenous Ags to the MHC class I pathway for induction of CTL response. PMID- 11046051 TI - Nonopsonic phagocytosis of Mycobacterium kansasii by human neutrophils depends on cholesterol and is mediated by CR3 associated with glycosylphosphatidylinositol anchored proteins. AB - Receptors involved in the phagocytosis of microorganisms under nonopsonic conditions have been little studied in neutrophils. Complement receptor type 3 (CR3) is a pattern recognition receptor able to internalize zymosan and C3bi coated particles. We report that Abs directed against CR3 strongly inhibited nonopsonic phagocytosis of Mycobacterium kansasii in human neutrophils. In these cells CR3 has been found associated with several GPI-anchored proteins localized in cholesterol-rich microdomains (rafts) of the plasma membrane. Cholesterol sequestration by nystatin, filipin, or beta-cyclodextrin as well as treatment of neutrophils with phosphatidylinositol phospholipase C to remove GPI-anchored proteins from the cell surface markedly inhibited phagocytosis of M. kansasii, without affecting phagocytosis of zymosan or serum-opsonized M. kansasii. Abs directed against several GPI-anchored proteins inhibited phagocytosis of M. kansasii, but not of zymosan. N:-acetyl-D-glucosamine, which is known to disrupt interactions between CR3 and GPI proteins, also strongly diminished phagocytosis of these mycobacteria. In conclusion, phagocytosis of M. kansasii involved CR3, GPI-anchored receptors, and cholesterol. In contrast, phagocytosis of zymosan or opsonized particles involved CR3, but not cholesterol or GPI proteins. We propose that CR3, when associated with a GPI protein, relocates in cholesterol-rich domains where M. kansasii are internalized. When CR3 is not associated with a GPI protein, it remains outside of these domains and mediates phagocytosis of zymosan and opsonized particles, but not of M. kansasii. PMID- 11046052 TI - MHC class Ib-restricted CTL provide protection against primary and secondary Listeria monocytogenes infection. AB - Infection of B6 mice with the intracellular pathogen Listeria monocytogenes (LM) results in the activation of CD8(+) T cells that respond to Ag presented by both MHC class Ia and class Ib molecules. Enzyme-linked immunospot analysis reveals that these CTL populations expand and contract at different times following a primary sublethal LM infection. Between days 4 and 6 postinfection, class Ib restricted CTL exhibit a rapid proliferative response that is primarily H2-M3 restricted. The peak response of class Ia-restricted CD8(+) T cells occurs a few days later, after the majority of bacteria have been cleared. Although class Ia restricted CTL exhibit a vigorous recall response to secondary LM infection, we observe limited expansion of class Ib-restricted memory CTL, even in MHC class Ia deficient mice (B6.K(b-/-)D(b-/-)). Despite this lack of enhanced expansion in vivo, class Ib-restricted memory CTL retain the ability to proliferate and expand when provided with Ag in vitro. Furthermore, we demonstrate that in vivo depletion of CD8(+) T cells in LM-immune B6.K(b-/-)D(b-/-) mice severely impairs memory protection. Together, these data demonstrate that class Ib-restricted CTL play an important role in clearing a primary LM infection and generate a memory population capable of providing significant protection against subsequent infection. PMID- 11046053 TI - Brucella abortus lipopolysaccharide in murine peritoneal macrophages acts as a down-regulator of T cell activation. AB - Macrophages play a central role in host immune responses against pathogens by acting as both professional phagocytic cells and as fully competent APCs. We report here that the LPS from the facultative intracellular Gram-negative bacteria Brucella abortus interferes with the MHC class II Ag presentation pathway. LPS inhibits the capacity of macrophages to present hen egg lysozyme (HEL) antigenic peptides to specific CD4(+) T cells but not those of OVA to specific CD8(+) T cells. This defect was neither related to a decrease of MHC class II surface expression nor to a deficient uptake or processing of HEL. In addition, B. abortus LPS did not prevent the formation of SDS-resistant MHC class II complexes induced by HEL peptides. At the cell surface of macrophages, we observed the presence of LPS macrodomains highly enriched in MHC class II molecules, which may be responsible for the significant down-regulation of CD4(+) T cell activation. This phenomenon may account for the avoidance of the immune system by certain bacterial pathogens and may explain the immunosuppression observed in individuals with chronic brucellosis. PMID- 11046054 TI - Role of p38 mitogen-activated protein kinase in rhinovirus-induced cytokine production by bronchial epithelial cells. AB - The stress-activated protein kinase p38 plays a central role in the regulation of cytokine biosynthesis by various cell types in response to a wide range of stimuli. Because the local inflammatory response and the infiltration of neutrophils is thought to contribute to the symptoms and sequelae of rhinovirus infection, we investigated the role of p38 kinase in cytokine and chemokine elaboration in airway epithelial cells infected with human rhinovirus. Rhinovirus 39 infection of BEAS-2B cells resulted in synthesis of cytokines (IL-1, IL-6, G CSF, and GM-CSF) and CXC chemokines (IL-8, epithelial neutrophil-activating protein-78, and growth-related oncogene-alpha), evident 24-72 h postinfection. Rhinovirus infection induced a time- and dose-dependent increase in tyrosine phosphorylation of p38 kinase, which peaked 30 min postinfection and remained elevated for 1 h. Treatment of infected cells with SB 239063, a potent pyridinyl imidazole inhibitor of p38 kinase, resulted in up to 100% inhibition of mediator production and partially reduced levels of IL-8 mRNA as determined by quantitative RT-PCR. Treatment with SB 239063 had no effect on virus replication and was not cytotoxic at concentrations 100-fold as compared with controls. Thus, through its activation of synovial fibroblasts, fibrin(ogen) deposition may promote the recruitment (via chemokines) and retention (via adhesion molecules) of lymphocytes within the arthritic joint. PMID- 11046060 TI - Regulation of endothelial CD73 by adenosine: paracrine pathway for enhanced endothelial barrier function. AB - During episodes of inflammation, multiple cell types release adenine nucleotides in the form of ATP, ADP, 5'-AMP, and adenosine. In particular, following activation, polymorphonuclear leukocytes release larger quantities of 5'-AMP. Extracellular 5'-AMP is metabolized to adenosine by surface-expressed 5' ectonucleotidase (CD73). Adenosine liberated by this process activates surface adenosine A(2B) receptors, results in endothelial junctional reorganization, and promotes barrier function. We hypothesized that adenosine signaling to endothelia provides a paracrine loop for regulated expression of CD73 and enhanced endothelial barrier function. Using an in vitro microvascular endothelial model, we investigated the influence of 5'-AMP; adenosine; and adenosine analogues on CD73 transcription, surface expression, and function. Initial experiments revealed that adenosine and adenosine analogues induce CD73 mRNA (RT-PCR), surface expression (immunoprecipitation of surface biotinylated CD73), and function (HPLC analysis of etheno-AMP conversion to ethenoadenosine) in a time- and concentration-dependent fashion. Subsequent studies revealed that similar exposure conditions increase surface protein through transcriptional induction of CD73. Analysis of DNA-binding activity by EMSA identified a functional role for CD73 cAMP response element and, moreover, indicated that multiple cAMP agonists induce transcriptional activation of functional CD73. Induced CD73 functioned to enhance 5'-AMP-mediated promotion of endothelial barrier (measured as a paracellular flux of 70-kDa FITC-labeled tracer). These results provide an example of transcriptional induction of enzyme (CD73) by enzymatic product (adenosine) and define a paracrine pathway for the regulated expression of vascular endothelial CD73 and barrier function. PMID- 11046061 TI - The CXC chemokine receptor 2, CXCR2, is the putative receptor for ELR+ CXC chemokine-induced angiogenic activity. AB - We have previously shown that members of the ELR(+) CXC chemokine family, including IL-8; growth-related oncogenes alpha, beta, and gamma; granulocyte chemotactic protein 2; and epithelial neutrophil-activating protein-78, can mediate angiogenesis in the absence of preceding inflammation. To date, the receptor on endothelial cells responsible for chemotaxis and neovascularization mediated by these ELR(+) CXC chemokines has not been determined. Because all ELR(+) CXC chemokines bind to CXC chemokine receptor 2 (CXCR2), we hypothesized that CXCR2 is the putative receptor for ELR(+) CXC chemokine-mediated angiogenesis. To test this postulate, we first determined whether cultured human microvascular endothelial cells expressed CXCR2. CXCR2 was detected in human microvascular endothelial cells at the protein level by both Western blot analysis and immunohistochemistry using polyclonal Abs specific for human CXCR2. To determine whether CXCR2 played a functional role in angiogenesis, we determined whether this receptor was involved in endothelial cell chemotaxis. We found that microvascular endothelial cell chemotaxis in response to ELR(+) CXC chemokines was inhibited by anti-CXCR2 Abs. In addition, endothelial cell chemotaxis in response to ELR(+) CXC chemokines was sensitive to pertussis toxin, suggesting a role for G protein-linked receptor mechanisms in this biological response. The importance of CXCR2 in mediating ELR(+) CXC chemokine-induced angiogenesis in vivo was also demonstrated by the lack of angiogenic activity induced by ELR(+) CXC chemokines in the presence of neutralizing Abs to CXCR2 in the rat corneal micropocket assay, or in the corneas of CXCR2(-/-) mice. We thus conclude that CXCR2 is the receptor responsible for ELR(+) CXC chemokine-mediated angiogenesis. PMID- 11046062 TI - A genome scan using a novel genetic cross identifies new susceptibility loci and traits in a mouse model of rheumatoid arthritis. AB - Proteoglycan-induced arthritis (PGIA) is a murine model for rheumatoid arthritis (RA) both in terms of its pathology and its genetics. PGIA can only be induced in susceptible mouse strains and their F(2) progeny. Using the F(2) hybrids resulting from an F(1) intercross of a newly identified susceptible (C3H/HeJCr) and an established resistant (C57BL/6) strain of mouse, our goals were to: 1) identify the strain-specific loci that confer PGIA susceptibility, 2) determine whether any pathophysiological parameters could be used as markers that distinguish between nonarthritic and arthritic mice, and 3) analyze the effect of the MHC haplotype on quantitative trait loci (QTL) detection. To identify QTLs, we performed a genome scan on the F(2) hybrids. For pathophysiological analyses, we measured pro- and antiinflammatory cytokines such as IL-1, IL-6, IFN-gamma, IL 4, IL-10, IL-12, Ag-specific T cell proliferation and IL-2 production, serum IgG1 and IgG2 levels of both auto- and heteroantibodies, and soluble CD44. We have identified four new PGIA-linked QTLs (Pgia13 through Pgia16) and confirmed two (Pgia5, Pgia10) from our previous study. All new MHC-independent QTLs were associated with either disease onset or severity. Comprehensive statistical analysis demonstrated that while soluble CD44, IL-6, and IgG1 vs. IgG2 heteroantibody levels differed significantly between the arthritic and nonarthritic groups, only Ab-related parameters colocalized with the QTLs. Importantly, the mixed haplotype (H-2(b) and H-2(k)) of the C3H x C57BL/6 F(2) intercross reduced the detection of several previously identified QTLs to suggestive levels, indicating a masking effect of unmatched MHCs. PMID- 11046063 TI - Transepithelial neutrophil migration is CXCR1 dependent in vitro and is defective in IL-8 receptor knockout mice. AB - Neutrophil migration across infected mucosal surfaces is chemokine dependent, but the role of chemokine receptors has not been investigated. In this study, chemokine receptors were shown to be expressed by epithelial cells lining the urinary tract, and to play an essential role for neutrophil migration across the mucosal barrier. Uroepithelial CXCR1 and CXCR2 expression was detected in human urinary tract biopsies, and in vitro infection of human uroepithelial cell lines caused a dramatic increase in both receptors. As a consequence, there was higher binding of IL-8 to the cells and the IL-8-dependent neutrophil migration across the infected epithelial cell layers was enhanced. Abs to IL-8 or to the CXCR1 receptor inhibited this increase by 60% (p<0.004), but anti-CXCR2 Abs had no effect, suggesting that CXCR1 was the more essential receptor in this process. Similar observations were made in the mouse urinary tract, where experimental infection stimulated epithelial expression of the murine IL-8 receptor, followed by a rapid flux of neutrophils into the lumen. IL-8 receptor knockout mice, in contrast, failed to express the receptor, their neutrophils were unable to cross the epithelial barrier, and accumulated in massive numbers in the tissues. These results demonstrate that epithelial cells express CXC receptors and that infection increases receptor expression. Furthermore, we show that CXCR1 is required for neutrophil migration across infected epithelial cell layers in vitro, and that the murine IL-8 receptor is needed for neutrophils to cross the infected mucosa of the urinary tract in vivo. PMID- 11046064 TI - Monocyte chemotactic protein-1 receptor CCR2B is a glycoprotein that has tyrosine sulfation in a conserved extracellular N-terminal region. AB - Monocyte chemotactic protein-1 (MCP-1) binding to its receptor, CCR2B, plays an important role in a variety of diseases involving infection, inflammation, and/or injury. In our effort to understand the molecular basis of this interaction and its biological consequences, we recognized a conserved hexad of amino acids at the N-terminal extracellular domain of several chemokine receptors, including CCR2B. Human embryonic kidney 293 cells expressing Flag-tagged CCR2B containing site-directed mutations in this region, 21-26, including a consensus tyrosine sulfation site were used to determine MCP-1 binding and its biological consequences. The results showed that several of these amino acids are important for MCP-1 binding and consequent lamellipodium formation, chemotaxis, and signal transduction involving adenylate cyclase inhibition and Ca(2+) influx into cytoplasm. Mutations that prevented adenylate cyclase inhibition and Ca(2+) influx did not significantly inhibit lamellipodium formation and chemotaxis, suggesting that these signaling events are not involved in chemotaxis. CCR2B was found to be sulfated at Tyr(26); this sulfation was abolished by the substitution of Tyr with Ala and severely reduced by substitution of Asp(25), a part of the consensus sulfation site. The expressed CCR2B was found to be N:-glycosylated, as N:-glycosidase F treatment of the receptor or growth of the cells in tunicamycin reduced the receptor size to the same level, from 50 to 45 kDa. Thus, CCR2B is the first member of the CC chemokine receptor family shown to be a glycoprotein that is sulfated at the N-terminal Tyr. These post-translational modifications probably have significant biological functions. PMID- 11046065 TI - Temporal development of autoreactive Th1 responses and endogenous presentation of self myelin epitopes by central nervous system-resident APCs in Theiler's virus infected mice. AB - Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease is a chronic-progressive, immune-mediated CNS demyelinating disease and a relevant model of multiple sclerosis. Myelin destruction is initiated by TMEV-specific CD4(+) T cells targeting persistently infected CNS-resident APCs leading to activation of myelin epitope-specific CD4(+) T cells via epitope spreading. We examined the temporal development of virus- and myelin-specific T cell responses and acquisition of virus and myelin epitopes by CNS-resident APCs during the chronic disease course. CD4(+) T cell responses to virus epitopes arise within 1 wk after infection and persist over a >300-day period. In contrast, myelin specific T cell responses are first apparent approximately 50-60 days postinfection, appear in an ordered progression associated with their relative encephalitogenic dominance, and also persist. Consistent with disease initiation by virus-specific CD4(+) T cells, CNS mononuclear cells from TMEV-infected SJL mice endogenously process and present virus epitopes throughout the disease course, while myelin epitopes are presented only after initiation of myelin damage (>50-60 days postinfection). Activated F4/80(+) APCs expressing high levels of MHC class II and B7 costimulatory molecules and ingested myelin debris chronically accumulate in the CNS. These results suggest a process of autoimmune induction in which virus-specific T cell-mediated bystander myelin destruction leads to the recruitment and activation of infiltrating and CNS-resident APCs that process and present endogenous myelin epitopes to autoreactive T cells in a hierarchical order. PMID- 11046066 TI - Human Peyer's patch T cells are sensitized to dietary antigen and display a Th cell type 1 cytokine profile. AB - Animal studies have demonstrated that feeding Ags induces regulatory (Th2, Th3) cells in Peyer's patches (PP), which migrate to the periphery and produce immunomodulatory cytokines such as IL-4, IL-10, or TGF-beta. In this work we have attempted to extend this paradigm to man by analyzing the response of human PP T cells to in vitro challenge with the common dietary Ag beta-lactoglobulin (betalg) of cow's milk. PP T cells stimulated with betalg showed enhanced proliferation compared with blood T cells from the same patient. Increased expression of CD25 and the Th1-associated chemokine receptor CCR5 was also seen on CD4(+) and CD8(+) PP T cells, but not blood T cells, stimulated with betalg. By enzyme-linked immunospot assay and RT-PCR, the PP T cell recall response to betalg and casein was dominated by IFN-gamma, with negligible IL-4, IL-5, IL-10, or TGF-beta. To help explain the PP T cell response to betalg, we examined IL-12 expression. Both IL-12p40 and -p35 transcripts were abundantly expressed in PP, but not in adjacent normal ileal mucosa. Immunoreactive IL-12p40-containing cells were present below the PP dome epithelium. Furthermore, in culture, PP, but not paired PBMC, spontaneously released IL-12p70. These results suggest that the human response to oral Ags in the gut may be different from that in rodents. PMID- 11046067 TI - Surges of increased T cell reactivity to an encephalitogenic region of myelin proteolipid protein occur more often in patients with multiple sclerosis than in healthy subjects. AB - We have previously shown that patients with multiple sclerosis (MS) have increased T cell responses to the immunodominant region (residues 184-209) of myelin proteolipid protein (PLP). The present study investigated whether this reactivity fluctuates over time and correlates with disease activity. We performed monthly limiting dilution assays for 12-16 mo in four healthy subjects and five patients with relapsing-remitting MS to quantify the frequencies of circulating T cells proliferating in response to PLP(41-58), PLP(184-199), PLP(190-209), myelin basic protein (MBP), MBP(82-100), and tetanus toxoid. Disease activity was monitored by clinical assessment and gadolinium-enhanced magnetic resonance imaging of the brain. There were fluctuations in the frequencies of autoreactive T cells in all subjects. Compared with healthy controls, MS patients had significantly more frequent surges of T cells reactive to the 184-209 region of PLP, but infrequent surges of T cell reactivity to MBP(82-100). There was temporal clustering of the surges of T cell reactivity to MBP(82-100) and MBP, suggesting T cell activation by environmental stimuli. Some clinical relapses were preceded by surges of T cell reactivity to PLP(184-209), and in one patient there was significant correlation between the frequency of T cells reactive to PLP(184-199) and the total number of gadolinium-enhancing magnetic resonance imaging lesions. However, other relapses were not associated with surges of T cell reactivity to the Ags tested. T cells reactive to PLP(184 209) may contribute to the development of some of the CNS lesions in MS. PMID- 11046068 TI - Up-regulation of IL-17 is associated with bioactive IL-8 expression in Helicobacter pylori-infected human gastric mucosa. AB - Helicobacter pylori (HP)-associated gastritis is characterized by an increased number of acute and chronic inflammatory cells secreting cytokines that contribute to maintain and expand the local inflammation. Locally induced IL-8 is believed to play a major role in the HP-associated acute inflammatory response. Factors/mechanisms that regulate IL-8 induction are, however, not fully understood. In the present study we investigated whether HP infection is associated with an increased production of IL-17, a T cell-derived cytokine capable of modulating IL-8 gene expression. We showed that both IL-17 RNA transcripts and protein were expressed at a higher level in the whole gastric mucosal and lamina propria mononuclear cell samples from HP-infected patients than in those from uninfected subjects. HP: eradication was associated with a marked down-regulation of IL-17 expression. The addition of a neutralizing anti IL-17 Ab to the gastric lamina propria mononuclear cell cultures resulted in a significant inhibition of IL-8 secretion, indicating that IL-17 contributes to enhance IL-8 in the HP-colonized gastric mucosa. Consistently, stimulation of MKN 28 cells, a gastric epithelial cell line, with IL-17 increased IL-8 secretion. Finally, conditioned medium from the IL-17-stimulated MKN 28 cell cultures promoted the in vitro polymorphonuclear leukocyte migration. This effect was inhibitable by a neutralizing IL-8 but not IL-17 Ab. Together, these data indicate that biologically active IL-17 production is increased during HP: infection, suggesting the possibility that this cytokine may play an important role in the inflammatory response to the HP colonization. PMID- 11046069 TI - Human double-negative T cells in systemic lupus erythematosus provide help for IgG and are restricted by CD1c. AB - To understand the mechanism of T cell help for IgG production in systemic lupus erythematosus (SLE) we investigated the response of CD4- and CD8-negative (double negative (DN)) T cells because 1) DN T cells are present at unusually high frequency in patients with SLE and can induce pathogenic autoantibodies; 2) the DN T cell repertoire includes cells restricted by CD1 Ag-presenting molecules; and 3) CD1c is expressed on a population of circulating B cells. We derived DN T cell lines from SLE patients and healthy individuals. In the presence of CD1(+) APCs, DN T cell lines from SLE patients produced both IL-4 and IFN-gamma, whereas DN T cells from healthy donors produced IFN-gamma, but no IL-4. In general, cells from patients with highly active disease produced high levels of IFN-gamma; cells from those with little activity produced high IL-4. Coculture of CD1c-directly reactive T cells from healthy donors with CD1c(+) B cells elicited IgM Abs, but little or no IgG. In contrast, CD1c-directly reactive T cells from SLE patients induced isotype switching, with a striking increase in IgG production. Neutralizing Abs to CD1c inhibited the ability of DN T cells to induce IgG production from CD1c(+) B cells, further indicating that CD1c mediated the T and B cell interaction. IgG production was also inhibited by neutralizing Abs to IL 4, correlating with the cytokine pattern of DN T cells derived from these patients. The data suggest that CD1c-restricted T cells from SLE patients can provide help to CD1c(+) B cells for IgG production and could therefore promote pathogenic autoantibody responses in SLE. PMID- 11046070 TI - Anti-SSA/Ro and anti-SSB/La autoantibodies bind the surface of apoptotic fetal cardiocytes and promote secretion of TNF-alpha by macrophages. AB - Despite the near universal association of congenital heart block and maternal Abs to SSA/Ro and SSB/La, the intracellular location of these Ags has made it difficult to substantiate their involvement in pathogenicity. To define whether components of the SSA/Ro-SSB/La complex, which translocate during apoptosis, are indeed accessible to extracellular Abs, two approaches were taken: immunoprecipitation of surface biotinylated proteins and scanning electron microscopy. Human fetal cardiocytes from 16-24-wk abortuses were cultured and incubated with staurosporine to induce apoptosis. Surface biotinylated 48-kDa SSB/La was reproducibly immunoprecipitated from apoptotic, but not nonapoptotic cardiocytes. Surface expression of SSA/Ro and SSB/La was further substantiated by scanning electron microscopy. Gold particles (following incubation with gold labeled sera containing various specificities of anti-SSA/Ro-SSB/La Abs and murine mAb to SSB/La and 60-kDa SSA/Ro) were consistently observed on early and late apoptotic cardiocytes. No particles were seen after incubation with control antisera. To evaluate whether opsonized apoptotic cardiocytes promote inflammation, cells were cocultured with macrophages. Compared with nonapoptotic cardiocytes or apoptotic cardiocytes incubated with normal sera, apoptotic cardiocytes preincubated with affinity-purified Abs to SSB/La, 52-kDa SSA/Ro, or 60-kDa SSA/Ro increased the secretion of TNF-alpha from cocultured macrophages. In summary, apoptosis results in surface accessibility of all SSA/Ro-SSB/La Ags for recognition by circulating maternal Abs. It is speculated that in vivo such opsonized apoptotic cardiocytes promote an inflammatory response by resident macrophages with damage to surrounding conducting tissue. PMID- 11046071 TI - Evidence for early aging in the mucosal immune system. AB - Despite recent advances in the cellular and molecular analysis of induction and regulation of mucosal immune responses, little is yet known about differences which occur in aging. To address this important issue, we have compared the mucosal and systemic immune responses of aged (12- to 14-mo- or 2-year-old) and young adult (6- to 8-wk-old) C57BL/6 mice. Both aged and young mice were immunized weekly with three oral doses of 1 mg of OVA and 10 microg of cholera toxin (CT) as mucosal adjuvant. Both groups of mice over 1 or 2 years of age showed reduced levels of Ag-specific mucosal or systemic immune responses at day 21. An Ag-specific B cell enzyme-linked immunospot assay confirmed these results at the cellular level. When the Ag-induced cytokine responses were examined at both protein and mRNA levels, CD4(+) T cells from spleen and Peyer's patches of young adult mice revealed elevated levels of IL-4 production; however, these cytokine responses were significantly diminished in aged mice. In contrast to mucosal immunization, mice s. c. immunized with OVA plus CT resulted in impaired OVA-specific but intact CT B subunit-specific immune responses in 12- to 14-mo old mice although the responses to both Ags were depressed in 2-year-old mice. These results provide the first evidence that the development of age-associated alterations possibly occurs earlier in the mucosal immune system than in the systemic immune compartment. PMID- 11046072 TI - Development of myelin oligodendrocyte glycoprotein autoreactive transgenic B lymphocytes: receptor editing in vivo after encounter of a self-antigen distinct from myelin oligodendrocyte glycoprotein. AB - We explored mechanisms involved in B cell self-tolerance against brain autoantigens in a double-transgenic mouse model carrying the Ig H-chain (introduced by gene replacement) and/or the L-chain kappa (conventional transgenic) of the mAb 8.18C5, specific for the myelin oligodendrocyte glycoprotein (MOG). Previously, we demonstrated that B cells expressing solely the MOG-specific Ig H-chain differentiate without tolerogenic censure. We show now that double-transgenic (THkappa(mog)) B cells expressing transgenic Ig H- and L-chains are subjected to receptor editing. We show that in adult mice carrying both MOG-specific Ig H- and L-chains, the frequency of MOG-binding B cells is not higher than in mice expressing solely the transgenic Ig H-chain. In fact, in THkappa(mog) double-transgenic mice, the transgenic kappa(mog) L-chain was commonly replaced by endogenous L-chains, i.e., by receptor editing. In rearrangement-deficient RAG-2(-) mice, differentiation of THkappa(mog) B cells is blocked at an immature stage (defined by the B220(low)IgM(low)IgD(-) phenotype), reflecting interaction of the autoreactive B cells with a local self-determinant. The tolerogenic structure in the bone marrow is not classical MOG, because back crossing THkappa(mog) mice into a MOG-deficient genetic background does not lead to an increase in the proportion of MOG-binding B cells. We propose that an as yet undefined self-Ag distinct from MOG cross-reacts with the THkappa(mog) B cell receptor and induces editing of the transgenic kappa(mog) L-chain in early immature B cells without affecting the pathogenic potential of the remaining MOG specific B cells. This phenomenon represents a particular form of chain-specific split tolerance. PMID- 11046073 TI - Molecular mechanisms of selective estrogen receptor modulator (SERM) action. AB - In females, estrogens play a key role in reproduction and have beneficial effects on the skeletal, cardiovascular, and central nervous systems. Most estrogenic responses are mediated by estrogen receptors (ERs), either ER alpha or ER beta, which are members of the nuclear receptor superfamily of ligand-dependent transcription factors. Selective estrogen receptor modulators (SERMs) are ER ligands that in some tissues act like estrogens, but block estrogen action in others. Thus, SERMs may exhibit an agonistic or antagonistic biocharacter depending on the context in which their activity is examined. For example, the SERMs tamoxifen and raloxifene both exhibit ER antagonist activity in breast and agonist activity in bone, but only tamoxifen manifests agonist activity in the uterus. Numerous studies have examined the molecular basis for SERM selectivity. Collectively they indicate that different ER ligands induce distinct structural changes in the receptor that influence its ability to interact with other proteins (e.g., coactivators or corepressors) critical for the regulation of target gene transcription. The relative expression of coactivators and corepressors, and the nature of the ER and of its target gene promoter affect SERM biocharacter. Taken together, SERM selectivity reflects the diversity of ER forms and coregulators, cell type differences in their expression, and the diversity of ER target genes. This model provides a basis for understanding the molecular mechanisms of SERM action, and should help identify new SERMs with enhanced tissue or target gene selectivity. PMID- 11046074 TI - Multiple central nervous system targets for eliciting beneficial effects on memory and cognition. AB - The development of drugs for the treatment of disorders of cognition has benefited from a more precise knowledge of the loss of specific neural pathways associated with certain neurodegenerative diseases such as Alzheimer's disease (AD). The loss of basal forebrain cholinergic neurons in AD has engendered the development of new compounds that target various aspects of the cholinergic system. However, limitations in the effectiveness of the most common of these, the anticholinesterases, have fueled the race to provide more efficacious compounds. In an attempt to avoid side effects and improve efficacy, other neuronal targets have been considered, including receptors for norepinephrine, dopamine, serotonin, excitatory amino acids, neural peptides, and others. Our laboratory has had the opportunity to study the memory-enhancing potential of many of the compounds developed expressly for these neuronal targets in macaques. Upon reviewing 21 such studies it was evident that: 1) To varying degrees, pharmacological manipulation of each target yielded improved task performance. 2) Combining pharmacological targets could lead to additive or synergistic effects on task performance. 3) Mature adult and aged monkeys provided equivalent estimates of drug effectiveness. 4) There appeared to be no limiting level of task improvement for compounds tested in aged and younger monkeys. 5) Certain of these agents also exhibited potential disease-modifying actions. Thus, certain memory-enhancing agents may prove more useful when implemented early in the course of a disease such as AD, and they also may enjoy a wide application for the treatment of the memory decline associated with normal aging. PMID- 11046075 TI - Induction of cyclooxygenase-2 in rat gastric mucosa by rebamipide, a mucoprotective agent. AB - Recent studies indicate an expression of mitogen-inducible cyclooxygenase (COX-2) in gastric mucosa. Rebamipide, a mucoprotective agent enhances prostaglandin (PG) synthesis. The present study was designed to clarify the mechanism for rebamipide induced mucosal protection. Male Sprague-Dawley rats were administered 5, 15, or 50 mg/kg/day rebamipide for 14 days. The expression of constitutive cyclooxygenase (COX-1) and COX-2 in gastric mucosa was determined using Western blot analysis. Another series of rats was used to examine 1) the levels of PGE(2) in stomach with and without pretreatment with a COX-2 inhibitor; 2) the protective action of rebamipide against gastric damage caused by 0.6 N HCl; and 3) the effects of a COX-2 inhibitor on rebamipide-induced gastric mucosal protection. COX-2 expression was enhanced, whereas COX-1 expression did not change significantly in the gastric mucosa of rats after treatment with rebamipide. The gastric mucosal PGE(2) was higher in the rebamipide groups than in the vehicle-treated group. Rebamipide also suppressed gastric damage induced by HCl in a dose-dependent manner. A COX-2 inhibitor blocked the rebamipide induced increase in mucosal PGE(2), and mucosal protection induced by rebamipide. The results indicate that rebamipide induces COX-2 expression, increases PGE(2) levels, and enhances gastric mucosal defense in a COX-2-dependent manner. Thus, COX-2 has an important role in the effects of rebamipide on gastric mucosal protection. PMID- 11046077 TI - Three-dimensional quantitative structure activity relationship computational approaches for prediction of human in vitro intrinsic clearance. AB - Future alternatives to the presently accepted in vitro paradigm of prediction of intrinsic clearance, which could be used earlier in the drug discovery process, would potentially accelerate efforts to identify better drug candidates with more favorable metabolic profiles and less likelihood of failure with regard to human pharmacokinetic attributes. In this study we describe two computational methods for modeling human microsomal and hepatocyte intrinsic clearance data derived from our laboratory and the literature, which utilize pharmacophore features or descriptors derived from molecular structure. Human microsomal intrinsic clearance data generated for 26 known therapeutic drugs were used to build computational models using commercially available software (Catalyst and Cerius(2)), after first converting the data to hepatocyte intrinsic clearance. The best Catalyst pharmacophore model gave an r of 0.77 for the observed versus predicted clearance. This pharmacophore was described by one hydrogen bond acceptor, two hydrophobic features, and one ring aromatic feature essential to discriminate between high and low intrinsic clearance. The Cerius(2) quantitative structure activity relationship (QSAR) model gave an r(2) = 0.68 for the observed versus predicted clearance and a cross-validated r(2) (q(2)) of 0.42. Similarly, literature data for human hepatocyte intrinsic clearance for 18 therapeutic drugs were also used to generate two separate models using the same computational approaches. The best Catalyst pharmacophore model gave an improved r of 0.87 and was described by two hydrogen bond acceptors, one hydrophobe, and 1 positive ionizable feature. The Cerius(2) QSAR gave an r(2) of 0.88 and a q(2) of 0.79. Each of these models was then used as a test set for prediction of the intrinsic clearance data in the other data set, with variable successes. These present models represent a preliminary application of QSAR software to modeling and prediction of human in vitro intrinsic clearance. PMID- 11046076 TI - Long-lasting facilitation of 4-amino-n-[2,3-(3)H]butyric acid ([(3)H]GABA) release from rat hippocampal slices by nicotinic receptor activation. AB - In this study we explored the effect of the stimulation of nicotinic acetylcholine receptors located on interneurons by measuring 4-amino-n-[2,3 (3)H]butyric acid ([(3)H]GABA) release and monitoring [Ca (2+)](i) in superfused hippocampal slices. In the presence of 6-cyano-7-nitroquinoxaline-2,3-dione, (+/ )-2-amino-5-phosphonopentanoic acid, and atropine, i.e., under the blockade of N methyl-D-aspartate and non-N-methyl-D-aspartate glutamate and muscarinic receptors, nicotine did not alter the spontaneous outflow of [(3)H]GABA, but significantly increased the stimulation-evoked [(3)H]GABA efflux. This effect of nicotine depended on the time interval between nicotine treatment and electrical stimulus, the concentration of nicotine (1-100 microM), and the parameters of electrical depolarization. Acetylcholine (0.03-3 mM), and the alpha 7 subtype selective agonist choline (0.1-10 mM), also potentiated stimulus-evoked release of [(3)H]GABA, whereas 1,1-dimethyl-4-phenilpiperazinium iodide failed to increase the tritium outflow significantly. The effect of nicotine treatment was prevented by tetrodotoxin (1 microM) and by the nicotinic acetylcholine receptor antagonist mecamylamine (10 microM), and the alpha 7 subtype-selective antagonists alpha-bungarotoxin (100 nM) and methyllycaconitine (10 nM), whereas dihidro-beta-erythroidine (20 nM) was without effect. Perfusion of 100 microM nicotine caused a [Ca(2+)](i) transient in about one-third of the tested interneurons; however, the response to subsequent electrical stimulation remained unchanged. Inhibition of the GABA transporter system by nipecotic acid (1 mM) or by decreasing the bath temperature to 12 degrees C abolished completely the effect of nicotine to potentiate the stimulation-evoked release of GABA. These findings indicate that the activation of alpha 7-type nicotinic receptors of hippocampal interneurons results in a long-lasting ability of these cells to respond to depolarization with an increased release of GABA mediated by the transporter system. PMID- 11046078 TI - The catalytic DNA topoisomerase II inhibitor dexrazoxane (ICRF-187) induces endopolyploidy in Chinese hamster ovary cells. AB - The bisdioxopiperazines, including dexrazoxane (ICRF-187), are catalytic or noncleavable complex-forming inhibitors of DNA topoisomerase II that do not produce DNA strand breaks. In this study we show that dexrazoxane inhibits the division of Chinese hamster ovary (CHO) cells resulting in marked increases in cell size (up to 80 microm in diameter), volume (up to 150-fold greater), and ploidy (as high as 32N). This last result indicates that the dexrazoxane-induced DNA reduplication was restricted to once per cell cycle. Kinetic analysis of the flow cytometry data indicated that the conversion between successively higher ploidy levels was progressively slowed at longer times of exposure to dexrazoxane. Both the protein and DNA content of dexrazoxane-treated CHO cells increased linearly over time in the same proportion. Light and electron microscopic studies of dexrazoxane-treated cells showed ring-like multilobulated nuclei. Immunohistochemical staining of dexrazoxane-treated cells showed that F actin and acetylated alpha-tubulin were present in large, highly organized networks. Immunohistochemical staining of the dexrazoxane-treated CHO cells also showed that the topoisomerase II alpha colocalized with the DNA of the multilobulated nuclei. Staining of gamma-tubulin revealed that the dexrazoxane treated cells contained multiple centrosomes, indicating that dexrazoxane prevents cytokinesis but not centrosome reduplication. It is concluded that dexrazoxane inhibits CHO cytokinesis in cells by virtue of its ability to inhibit topoisomerase II. PMID- 11046079 TI - Enhanced endothelin(A) receptor-mediated calcium mobilization and contraction in organ cultured porcine coronary arteries. AB - Arterial injury models for coronary artery disease have demonstrated an enhanced expression and function of either the endothelin(A) or endothelin(B) (ET(A) or ET(B)) receptor subtype. We hypothesized that organ culture would enhance the physiological function of ET receptors in the porcine right coronary artery. Arteries were either cold stored (4 degrees C) or organ cultured (37 degrees C) for 4 days. After 4 days, the artery was either 1) sectioned into rings to measure the ET-1-induced isometric tension response (3 x 10(-10)-3 x 10(-7) M), or 2) enzymatically dispersed and the isolated smooth muscle cells imaged using fura-2 to measure the myoplasmic calcium (Ca(m)) response to 3 x 10(-8) M ET-1 ( approximately EC(50)). Isometric tension and Ca(m) to ET-1 were measured in the absence and presence of bosentan (nonselective ET(A) or ET(B) receptor antagonist), BQ788 (ET(B)-selective antagonist), and BQ123 (ET(A)-selective antagonist). Compared with cold storage, organ culture induced a 2-fold increase in tension development (3 x 10(-7) M ET-1) and Ca(m) (3 x 10(-8) M ET-1), which was inhibited with bosentan, thus confirming the enhanced responses to ET-1 were due to ET receptor activation. BQ123 also inhibited the enhanced contraction and Ca(m) responses to ET-1. In contrast, BQ788 failed to inhibit tension development and Ca(m) responses to ET-1 in organ culture and cold storage. Sarafotoxin 6C (ET(B) agonist) failed to elicit an increased Ca(m) response in organ culture compared with cold storage. Our results indicate the increased tension development and Ca(m) responses to ET-1 in organ culture are attributable to ET(A) receptors, and not ET(B) receptors. PMID- 11046080 TI - Prevention of arterial thrombosis by intravenously administered platelet P2T receptor antagonist AR-C69931MX in a canine model. AB - P2Y1, P2X1, and P2T receptors mediate ADP-induced platelet aggregation. The antithrombotic effects of AR-C69931MX (N6-[2-methylthio)ethyl]-2-[3,3,3 trifluoropropylthio]-5'-adenylic acid, monoanhydride with dichloromethylenebiphosphonic acid), a selective P2T platelet receptor antagonist, was assessed in a canine model of arterial thrombosis. Placebo or AR C69931MX (4.0 microg/kg/min for 6 h) pretreatment was administered as an intravenous infusion beginning 15 min before inducing vessel wall injury. A 300 microA anodal current was applied to the intima of the carotid artery for 180 min or discontinued 30 min after cessation of blood flow due to thrombus formation. Each of five control animals developed occlusive thrombi within 3 h after induction of vessel wall injury. In contrast, carotid artery blood flow in five of six AR-C69931MX-treated animals was maintained for the duration of the protocol. Ex vivo platelet aggregation in response to adenosine diphosphate was inhibited at the first measurement time point of 75 min after the start of drug infusion and remained inhibited during drug administration. Bleeding time values were increased in the drug-treated group. Values for both the ex vivo platelet aggregation and the bleeding times returned to control values shortly after discontinuation of AR-C69931MX. The results indicate that AR-C69931MX antagonizes the ex vivo and in vivo aggregatory actions of ADP, and displays a rapid onset and offset of action with the ability to prevent occlusive arterial thrombus formation. AR-C69931MX may be suitable for the management of patients who require short-term modulation of platelet function. PMID- 11046081 TI - Role of inducible nitric-oxide synthase in regulation of whole-cell current in lung epithelial cells. AB - Lung inflammation is associated with enhanced expression of proinflammatory cytokines and increased production of nitric oxide (NO) by inducible NO synthase (iNOS). To investigate the possible relationship between cytokine-induced expression of iNOS and epithelial ion channel function, we measured whole-cell current in A549 cells treated with a mixture of cytokines: tumor necrosis factor, interleukin-1 beta, and interferon-gamma for 12 h. Cytokines significantly increased the expression and activity of iNOS, and reduced generation of cGMP in response to stimulation with NO donor S-nitroso-glutathione (GSNO). Patch-clamp studies showed that 100 microM GSNO increased the whole-cell current from 11.2 +/ 1.8 to 19.6 +/- 2.7 pA/pF (n = 16) in control cells, but had no effect in cytokine-treated cells (n = 9). N-(3-(Aminomethyl)benzyl)acetamidine (1400W), a selective inhibitor of iNOS, restored activation of the current by GSNO in cytokine-treated cells, indicating a crucial role for iNOS in this process. Cells treated with cytokines showed increased levels of peroxynitrite (ONOO(-)), compared with the control, or cells that were treated with the cytokines and 1400W or superoxide dismutase/catalase. Treatment of cells with 100 microM ONOO( ) had no effect on the whole-cell current, but in contrast to untreated cells, subsequent application of GSNO did not activate the current. In conclusion, cytokine-induced expression of iNOS affects activation of the whole-cell current via NO/cGMP pathway, likely by increasing the generation of ONOO(-). PMID- 11046082 TI - Excitatory and inhibitory actions of isoprostanes in human and canine airway smooth muscle. AB - Isoprostanes are generated nonenzymatically during free radical-mediated lipid peroxidation, and are used clinically and experimentally as markers of oxidative stress. However, their biological effects are poorly understood. We examined the effects of seven different 8-isoprostanes in human and canine airway smooth muscles. In large order airways (carina) of the human, several isoprostanes evoked powerful contractions, with 8-iso-prostaglandin (PG) E(2), 8-iso-PGF(1 alpha), and 8-iso-PGF(2 alpha) being the most efficacious (and with logEC(50) values of 7.0, 5.9, and 6.2 microM, respectively). These contractions were sensitive to the prostanoid TP receptor antagonist ICI 192,605 (0.1-1 microM), but not the EP prostanoid receptor antagonist AH-6809 (50 microM), or the leukotriene receptor antagonists monteleukast or ICI 198,615 (both 1 microM). Qualitatively similar results were obtained in small order human airways (<2 mm o.d.), except that the isoprostanes were generally slightly less potent. None of the isoprostanes had any marked excitatory effect in canine airways. In carbachol preconstricted tissues (pretreated with ICI 192,605 to block any potential contraction), several isoprostanes completely relaxed canine airways: 8-iso PGE(1), 8-iso-PGE(2), and 8-iso-PGF(3 alpha) were the most potent, with logIC(50) values of 6.9, 6.9, and 5.7, respectively. Only 8-iso-PGF(3 alpha) relaxed human airways (logIC(50) = 4.9). Our results show that several 8-isoprostanes are highly biologically active in human and canine airways, evoking both excitatory and/or inhibitory effects, and that these effects are compound, species, and tissue dependent. PMID- 11046083 TI - Altered hepatobiliary disposition of acetaminophen glucuronide in isolated perfused livers from multidrug resistance-associated protein 2-deficient TR(-) rats. AB - Previous studies have demonstrated that phenobarbital treatment impairs the biliary excretion of acetaminophen glucuronide (AG), although the transport system(s) responsible for AG excretion into bile has not been identified. Initial studies in rat canalicular liver plasma membrane vesicles indicated that AG uptake was stimulated modestly by ATP, but not by membrane potential, HCO(3)(-), or pH gradients. To examine the role of the ATP-dependent canalicular transporter multidrug resistance-associated protein 2 (Mrp2)/canalicular multispecific organic anion transporter (cMOAT) in the biliary excretion of AG, the hepatobiliary disposition of acetaminophen, AG, and acetaminophen sulfate (AS) was examined in isolated perfused livers from control and TR(-) (Mrp2-deficient) Wistar rats. Mean bile flow in TR(-) livers was approximately 0.3 microl/min/g of liver ( approximately 4-fold lower than control). AG biliary excretion was decreased (>300-fold) to negligible levels in TR(-) rat livers, indicating that AG is an Mrp2 substrate. Similarly, AS biliary excretion in TR(-) livers was decreased ( approximately 5-fold); however, concentrations were still measurable, suggesting that multiple mechanisms, including Mrp2-mediated active transport, may be involved in AS biliary excretion. AG and AS perfusate concentrations were significantly higher in livers from TR(-) compared with control rats. Pharmacokinetic modeling of the data revealed that the rate constant for basolateral egress of AG increased significantly from 0.028 to 0.206 min(-1), consistent with up-regulation of a basolateral organic anion transporter in Mrp2 deficient rat livers. In conclusion, these data indicate that AG biliary excretion is mediated by Mrp2, and clearly demonstrate that substrate disposition may be influenced by alterations in complementary transport systems in transport deficient animals. PMID- 11046084 TI - Tamoxifen acutely relaxes coronary arteries by an endothelium-, nitric oxide-, and estrogen receptor-dependent mechanism. AB - In epidemiological studies tamoxifen has been associated with a reduction in the incidence of fatal myocardial infarction in women. However, the effects of tamoxifen on coronary artery reactivity are unknown. We hypothesized that tamoxifen would relax precontracted isolated rabbit coronary arteries. Rings of coronary artery from adult male and nonpregnant female New Zealand White rabbits were suspended in organ baths containing Krebs' solution; isometric tension was measured. Tamoxifen (0.1-100 microM) induced significant endothelium-dependent relaxation in precontracted rabbit coronary arteries. This relaxation was inhibited by N(omega)-nitro-L-arginine methyl ester and the estrogen receptor antagonist ICI 182,780. There was no significant effect on calcium concentration dependent contraction curves. These data suggest that tamoxifen has beneficial effects on coronary reactivity that could, at least in part, account for the reduction in risk of fatal myocardial infarction in women taking tamoxifen. PMID- 11046085 TI - Effects of tetraethylammonium analogs on apoptosis and membrane currents in cultured cortical neurons. AB - Tetraethylammonium (TEA), the quaternary ammonium ion and nonselective K(+) channel blocker, is protective against neuronal apoptosis. We now tested two TEA analogs, tetrapentylammonium (TPeA) and tetrahexylammonium (THA), for their effects on apoptotic neuronal death and for their pharmacological profiles on membrane currents in cultured mouse cortical neurons. TPeA and THA (0.1-1.0 microM) attenuated staurosporine-induced caspase-3 activation and neuronal apoptosis. TPeA and THA blocked the outward delayed rectifier K(+) (I(K)) current in concentration-dependent manners with IC(50) values of 2.7 and 1.9 microM, respectively. I(K) was blocked by TPeA in a use-dependent manner, whereas THA blocked I(K) regardless of activation state of the channel. TPeA at 1 microM inhibited the high voltage-activated (HVA) Ca(2+) current and the A-type K(+) current (I(A)). TPeA (1-10 microM) also blocked the fast inactivating Na(+) current. The ligand-gated N-methyl-D-aspartate (NMDA) receptor current was not affected by up to 20 microM TPeA. THA at 1 microM showed inhibitory effects on I(A), HVA Ca(2+), and Na(+) currents. THA (10 microM) suppressed NMDA currents. The data suggest that, as K(+) channel blockers and apoptosis antagonists, TPeA and THA are much more potent than TEA; however, they have nonspecific actions on several voltage-gated or ligand-gated channels. PMID- 11046086 TI - Site of action of lubeluzole on voltage-sensitive Ca(2+) channels in isolated dorsal root ganglion cells of the rat: influence of pH. AB - Besides other pharmacological effects, the neuroprotective compound lubeluzole blocks low-voltage-activated (iLVA) and high-voltage-activated (iHVA) calcium channel currents. We investigated the site of action of lubeluzole on Ca(2+) channels in isolated dorsal root ganglion cells of the rat, using whole-cell voltage clamp. Experiments with extracellular application of 3 microM lubeluzole (pK(a) = 7.6) at different values of extracellular pH suggest that the protonated form of lubeluzole contributes to the block of iLVA and iHVA from the extracellular side. The partial block of iLVA and iHVA by 3 microM lubeluzole at extracellular pH 9 and intracellular pH (pH(i)) 9 indicates that the uncharged form of lubeluzole (L) may contribute to the block as well. The voltage-dependent acceleration of the apparent inactivation of iHVA by lubeluzole was much more pronounced at lower pH(i), which is consistent with membrane penetration of L and an open channel block of iHVA by the prononated form of lubeluzole acting from the intracellular side. Decreasing pH(i) induced a negative shift of the half inactivation potential of iLVA and increased the lubeluzole-induced block of iLVA. Experiments with extracellular or intracellular application of a quaternary ammonium derivative of lubeluzole (R133121), which was less potent than lubeluzole, support the above conclusions on the side of action of lubeluzole. Application of lubeluzole via the patch pipette affected iLVA and iHVA only minimally compared with extracellular application, probably partly due to efflux of L through the cell membrane. These experiments suggest that lubeluzole blocks Ca(2+) channels from both the extracellular and the intracellular side. PMID- 11046087 TI - Analgesic synergy between topical lidocaine and topical opioids. AB - Topical drugs avoid many of the problematic side effects of systemic agents. Immersion of the tail of a mouse into a solution of dimethyl sulfoxide (DMSO) containing morphine produces a dose-dependent, naloxone-sensitive, analgesia (ED(50) 6.1 mM; CL 4.3, 8.4) limited to the portion of the tail exposed to the drug. DMSO alone in this paradigm had no analgesic activity. Like morphine, the opioids levorphanol (ED(50) 5.0 mM; CL 3.8, 7.8) and buprenorphine (ED(50) 1. 1 mM; CL 0.7, 1.5) were effective topical analgesics. Lidocaine also was active in the tail-flick assay (ED(50) 2.5 mM; CL 2.0, 3.4), with a potency greater than morphine. As expected, the free base of lidocaine was more potent than its salt. Combinations of a low dose of lidocaine with a low dose of an opioid yielded significantly greater than additive effects for all opioids tested. Isobolographic analysis confirmed the presence of synergy between lidocaine and morphine, levorphanol and buprenorphine. These studies demonstrate a potent interaction peripherally between opioids and a local anesthetic and offer potential advantages in the clinical management of pain. PMID- 11046088 TI - delta-Opioid receptors are more efficiently coupled to adenylyl cyclase than to L type Ca(2+) channels in transfected rat pituitary cells. AB - Opioid receptors often couple to multiple effectors within the same cell. To examine potential mechanisms that contribute to the specificity by which delta receptors couple to distinct intracellular effectors, we stably transfected rat pituitary GH(3) cells with cDNAs encoding for delta-opioid receptors. In cells transfected with a relatively low delta-receptor density of 0.55 pmol/mg of protein (GH(3)DOR), activation of delta-receptors produced inhibition of adenylyl cyclase activity but was unable to alter L-type Ca(2+) current. In contrast, activation of delta-receptors in a clone that contained a higher density of delta receptors (2.45 pmol/mg of protein) and was also coexpressed with mu-opioid receptors (GH(3)MORDOR), resulted in not only the expected inhibition of adenylyl cyclase activity but also produced inhibition of L-type Ca(2+) current. The purpose of the present study was to determine whether these observations resulted from differences in delta-opioid receptor density between clones or interaction between delta- and mu-opioid receptors to allow the activation of different G proteins and signaling to Ca(2+) channels. Using the delta-opioid receptor alkylating agent SUPERFIT, reduction of available delta-opioid receptors in GH(3)MORDOR cells to a density similar to that of delta-opioid receptors in the GH(3)DOR clone resulted in abolishment of coupling to Ca(2+) channels, but not to adenylyl cyclase. Furthermore, although significantly greater amounts of all G proteins were activated by delta-opioid receptors in GH(3)MORDOR cells, delta opioid receptor activation in GH(3)DOR cells resulted in coupling to the identical pattern of G proteins seen in GH(3)MORDOR cells. These findings suggest that different threshold densities of delta-opioid receptors are required to activate critical amounts of G proteins needed to produce coupling to specific effectors and that delta-opioid receptors couple more efficiently to adenylyl cyclase than to L-type Ca(2+) channels. PMID- 11046089 TI - A pyrroline derivative of mexiletine offers marked protection against ischemia/reperfusion-induced myocardial contractile dysfunction. AB - The efficacy and mechanism of protection of a new 2,2,5, 5-tetramethylpyrroline derivative of mexiletine, MEX-NH, against ischemia/reperfusion-induced cardiac dysfunction are reported. The MEX-NH and its nitroxide metabolite are membrane permeable antioxidants. Studies were performed in an isolated rat heart model to measure the efficacy of MEX-NH in preventing postischemic injury. Serial measurements of contractile function and coronary flow were performed on hearts subjected to 30 min of global 37 degrees C ischemia followed by 45 min of reperfusion. Hearts were either untreated or treated with 25 microM MEX-NH or MEX for 1 min before ischemia. The hearts treated with MEX-NH showed marked recovery of left ventricular developed pressure (96.3 +/- 2.7% of preischemic value) compared with untreated (13.7 +/- 1.0%) or MEX-treated (19.9 +/- 2.7%) hearts. The cardiac sarcolemmal Na(+),K(+)-ATPase activity showed that the enzyme activity was fully restored in hearts treated with MEX-NH compared with 65 +/- 5.3% inhibition in the untreated hearts. Competitive inhibition of [(3)H]ouabain binding revealed that the MEX-NH binds at the K(+)-binding site of the enzyme. The present study establishes that the compound MEX-NH provides marked protection against ischemia/reperfusion-induced contractile dysfunction in isolated hearts. A combination of reversible inhibition of Na(+)/K(+)-ATPase activity during ischemia and site-targeted antioxidative effect upon reperfusion seems to contribute to this cardioprotection. PMID- 11046090 TI - Alterations of N-methyl-D-aspartate receptor properties after chemical ischemia. AB - Sublethal ischemic challenges can protect neurons against a second, more severe hypoxic insult. We report here that nonlethal chemical ischemia induces a transient alteration of NMDA receptors in rat cortical neurons in culture. Cells were incubated with 3 mM KCN in a glucose-free solution for 90 min. Analysis of NMDA receptor unitary events in patches excised from KCN-treated neurons showed an increased incidence of a small conductance channel 24 h after chemical ischemia. Whole-cell recordings of NMDA-induced currents 1 day after cyanide exposure revealed a significant increase in voltage-dependent extracellular Mg(2+) block compared with untreated neurons. The block reverted to control levels within 48 h. Both of these changes in the NMDA receptor could decrease the overall current flowing through the channel. Message levels for the NMDA receptor subunits NR1, NR2A, and NR2B were not different between the chemically challenged neurons and control cells, whereas NR2C message was barely detectable in either group. These results suggest that the alterations in NMDA receptor properties after KCN exposure may contribute to the molecular mechanisms that are activated in neurons to withstand lethal ischemic events in the brain after preconditioning. PMID- 11046091 TI - Endothelin receptor antagonism does not prevent the development of in vivo glyceryl trinitrate tolerance in the rat. AB - There is evidence that increased endothelial production of endothelin-1 (ET-1) may contribute to glyceryl trinitrate (GTN) tolerance. We used the competitive ET(A) receptor antagonist ZD2574 to determine whether chronic ET(A) receptor blockade affected the biochemical and functional responses to GTN during the development of GTN tolerance in vivo. Tolerance induced using transdermal GTN patches resulted in a 5.3 +/- 1.2-fold increase in the EC(50) value for GTN relaxation in isolated aorta from GTN-tolerant rats. Coadministration of ZD2574 (100 mg kg(-1) t.i.d. for 3 days) during tolerance induction had no effect on GTN induced relaxation. This dose of ZD2574 markedly blunted the pressor response to ET-1, indicating effective blockade of ET(A) receptors, and also abolished the initial transient depressor response to ET-1, indicating that blockade of endothelial ET(B) receptors also occurred using this dosage regimen for ZD2574. Consistent with the relaxation data, coadministration of ZD2574 had no effect on the decrease in GTN-induced cGMP accumulation or on the decrease in GTN biotransformation that occurred in aortae from GTN-tolerant animals. Radioimmunoassay data indicated that the GTN tolerance induction protocol caused a 2.3 +/- 0.4-fold and a 2.2 +/- 0.5-fold increase in total tissue ET-1 levels in tolerant aorta and vena cava, respectively. These data suggest that chronic inhibition of ET receptors by ZD2574 was not sufficient to prevent or diminish the tolerance-inducing effects of GTN, and that the increase in ET-1 levels observed in tolerant tissues may occur as a consequence of the vascular changes that occur during chronic GTN exposure. PMID- 11046092 TI - Rationale for the combination of PGE(1) and S-nitroso-glutathione to induce relaxation of human penile smooth muscle. AB - Many men with erectile dysfunction have been successfully treated with intracavernosal injection of prostaglandin E(1) (PGE(1)) but this treatment is ineffective in 30 to 40% of patients. The goals of this study were to characterize PGE(1)-induced relaxation of isolated human penile smooth muscle (penile arteries and trabecular strips), correlating this in vitro response with the clinical response to this drug, and to evaluate the effects of the combination of PGE(1) with S-nitrosoglutathione (SNO-Glu) on relaxation of isolated human penile smooth muscle. Large variability in the EC(50) and maximal relaxation induced by PGE(1) was observed between tissues of different patients. Patients with poor clinical response to intracavernosal alprostadil (PGE(1)) had significantly larger EC(50) values and smaller maximal relaxation compared with patients with partial or complete clinical response to this drug. SNO-Glu consistently produced complete or near complete relaxation of human corpus cavernosum strips and penile arteries, even when the tissue responded poorly to PGE(1). In trabecular strips, the combination of PGE(1) and SNO-Glu in a 1:100 ratio demonstrated a synergistic relaxation effect. The combination of PGE(1) and SNO-Glu simultaneously increased the levels of both cAMP and cGMP in human corpus cavernosum tissue. Our results suggest that the clinical effectiveness of intracavernosal administration of PGE(1) is related to the variability of the relaxation responses of human trabecular tissue and penile arteries to this drug. The synergistic interaction of PGE(1) and SNO-Glu makes this combination an effective method to cause penile smooth muscle relaxation, a necessary step to initiate and maintain penile erection. PMID- 11046093 TI - Hypoglycemic effect of insulin-transferrin conjugate in streptozotocin-induced diabetic rats. AB - Transferrin (Tf) receptor-mediated transcytosis of insulin-transferrin conjugate (In-Tf) has been demonstrated in cultured human enterocyte-like Caco-2 cells. In the present report, oral delivery of insulin as a Tf conjugate in streptozotocin (STZ)-induced diabetic rats was investigated. Human insulin was conjugated at a 1:1 molar ratio to iron-loaded human Tf by a disulfide linkage. The stability of In-Tf and the free insulin released from In-Tf was studied in the presence of rat liver slices by using radioimmunoassay. The release of free insulin involved a disulfide reduction reaction that was inhibited by the pretreatment of the liver slice with a sulfhydryl-reactive reagent N-ethylmaleimide. A protease inhibitor cocktail also showed a partial inhibition of insulin degradation. The biological activity of the conjugate was tested in STZ-induced diabetic rats with s.c. administration, and the conjugate exhibited a slow but prolonged hypoglycemic effect compared with that of the native human insulin. In-Tf also displayed a slow but prolonged hypoglycemic effect after oral administration in fasted STZ induced diabetic rats in a dose-dependent manner. Furthermore, In-Tf was detected in the serum of rats at 4 h after oral administration of the conjugate, indicating that In-Tf can overcome the barriers in the gastrointestinal tract and be absorbed as an intact conjugate. These results demonstrate that transepithelial transport via TfR-mediated transcytosis is a feasible approach for developing the oral delivery of insulin, as well as other peptide drugs. PMID- 11046094 TI - Modulation of nitric-oxide synthase by nicotine. AB - Effects of nicotine on arterial endothelium-dependent relaxations mediated by nitric oxide are controversial. Experiments were designed to test the hypothesis that nicotine can directly alter activity of endothelial nitric-oxide synthase (eNOS). NOS from aortic endothelial cells of untreated dogs and recombinant eNOS, neuronal NOS, and inducible NOS were used for these experiments. NOS activity was determined as conversion of L-[(3)H]arginine to L-[(3)H]citrulline in the absence or presence of nicotine (10(-7)-10(-3) M) in vitro. In separate assays, concentrations of cofactors NADPH, FAD, and tetrahydrobioprotein were reduced by half to assess for possible interaction with nicotine. With enzyme from aortic endothelial cells, total and calcium-dependent accumulation of citrulline increased by 30% in the presence of 10(-5) M nicotine. Nicotine dose dependently also increased citrulline accumulation by recombinant eNOS and neuronal NOS but not inducible NOS. Effects of nicotine on accumulation of citrulline by isolated eNOS and recombinant eNOS were further modulated by changes in the concentration of NADPH in the incubation solution. Our data demonstrate a significant effect of nicotine on eNOS-mediated citrulline accumulation. These results suggest that effects of nicotine on production of nitric oxide may depend on NADPH or oxygen radical interactions with NOS and thus may explain, in part, inconsistent findings of changes in production of endothelium-derived nitric oxide with nicotine administration. PMID- 11046095 TI - Signaling mechanisms coupled to presynaptic A(1)- and H(3)-receptors in the inhibition of cholinergic contractile responses of the guinea pig ileum. AB - The mechanisms coupled to adenosine A(1)- and histamine H(3)-receptors have been examined in the presynaptic inhibition of acetylcholine (ACh) release from the guinea pig ileum. Electrically evoked twitch contractions were used as a measure of neuronal ACh release. A(1)- and H(3)-receptors were activated by adenosine and R-(alpha)-methylhistamine (RAMH), respectively. The neuroinhibitory effect of adenosine and RAMH was augmented in the presence of the N-type Ca(2+) channel blocker, omega-conotoxin GVIA but unaffected by the L-type Ca(2+) channel blocker, nifedipine. The irreversible adenylyl cyclase inhibitor, MDL-12330A, potentiated the action of both adenosine and RAMH. Conversely, neither agonist was affected by the cAMP phosphodiesterase III and IV inhibitors, SKF-95654 and Ro-20-1724, respectively, or the cAMP antagonist, (R(p))-adenosine 3',5'-cyclic monophosphorothioate triethylamine. The neuromodulatory effect of adenosine, only, was potentiated by the cGMP phosphodiesterase V inhibitors, SKF-96231 and 1,3-dimethyl-6-(2-propoxy-5-methanesulfonylamidophenyl)- pyrazolo[3, 4 d]pyrimidin-4-(5H)-one but was unmodified by the cGMP analog, 8-bromo-cGMP or the guanylyl cyclase inhibitors, N-methylhydroxylamine and 1H-[1,2,4]oxadiazolo[4, 3 a]quinoxaline-1-one (ODQ). N-Methylhydroxylamine reduced, and ODQ potentiated, the inhibitory action of H(3)-receptor activation, but 8-bromo-cGMP was without effect. The study suggests that presynaptic A(1)- and H(3)-receptors inhibit cholinergic neurotransmission in the guinea pig ileum by limiting the availability of intraneuronal Ca(2+) via inhibition of N-type Ca(2+) channels. The balance of evidence does not support the involvement of the adenylyl cyclase/cAMP or guanylyl cyclase/cGMP systems. PMID- 11046096 TI - Interactions of the 5-hydroxytryptamine 3 antagonist class of antiemetic drugs with human cardiac ion channels. AB - Administration of the 5-hydroxytryptamine 3 receptor class of antiemetic agents has been associated with prolongation in the QRS, JT, and QT intervals of the ECG. To explore the mechanisms underlying these findings, we examined the effects of granisetron, ondansetron, dolasetron, and the active metabolite of dolasetron MDL 74,156 on the cloned human cardiac Na(+) channel hH1 and the human cardiac K(+) channel HERG and the slow delayed rectifier K(+) channel KvLQT1/minK. Using patch-clamp electrophysiology we found that all of the drugs blocked Na(+) channels in a frequency-dependent manner. At a frequency of 3 Hz, the IC(50) values for block of Na(+) current measured 2.6, 88.5, 38.0, and 8.5 microM for granisetron, ondansetron, dolasetron, and MDL 74,156, respectively. Block was relieved by strong hyperpolarizing potentials, suggesting a possible interaction with an inactivated channel state. Recovery from inactivation was impaired at -80 mV compared with -100 mV, and the fractional recovery was impaired by drug in a concentration-dependent manner. IC(50) values for block of the HERG cardiac K(+) channel measured 3.73, 0.81, 5.95, and 12.1 microM for granisetron, ondansetron, dolasetron, and MDL 74,156, respectively. Ondansetron (3 microM) also slowed decay of HERG tail currents. In contrast, none of these drugs (10 microM) produced greater than 30% block of the slow delayed rectifier K(+) channel KvLQT1/minK. We concluded that the antiemetic agents tested in this study block human cardiac Na(+) channels probably by interacting with the inactivated state. This may lead to clinically relevant Na(+) channel blockade, especially when high heart rates or depolarized/ischemic tissue is present. The submicromolar affinity of ondansetron for the HERG K(+) channel likely underlies the prolongation of cardiac repolarization reported for this drug. PMID- 11046097 TI - Protective effect of omapatrilat, a vasopeptidase inhibitor, on the metabolism of bradykinin in normal and failing human hearts. AB - Because part of the cardioprotective effects of angiotensin-converting enzyme (ACE) inhibitors results from their protective effects on cardiac bradykinin (BK) metabolism, the purpose of this study was to define the metabolism of BK in normal and failing human hearts and to compare the effect of omapatrilat, a vasopeptidase inhibitor (VPI), which simultaneously inhibits both neutral endopeptidase (NEP) and ACE, with that of an ACE inhibitor. Exogenous BK at a nanomolar concentration was incubated alone, in the presence of an ACE inhibitor (ramiprilat, 36 nM), or in the presence of a VPI (omapatrilat, 61 nM) with left ventricular membranes prepared from normal donor hearts (n = 7), and hearts from patients with an ischemic (n = 11) or dilated (n = 12) cardiomyopathy (DCM). The half-lives calculated for BK alone (199 +/- 60, 224 +/- 108, and 283 +/- 122 s; P = NS) exhibited similar values for normal, ischemic, and DCM heart tissues, respectively. Ramiprilat significantly increased the half-life of BK (P <.01), but the effect was similar for the three kinds of tissues (297 +/- 104, 267 +/- 157, and 407 +/- 146 s, respectively; P = NS). The potentiating effect of the VPI omapatrilat on the kinetic parameter of BK (478 +/- 210, 544 +/- 249, and 811 +/- 349 s, respectively) was greater than that of the ACE inhibitor (P <.01). Moreover, omapatrilat had a more important potentiating effect with DCM than normal heart membranes (P <.05). These results show that not only ACE but also and mainly NEP play an important role in the degradation of BK in human heart membranes. Omapatrilat, a VPI, has a greater protective effect on BK metabolism than that of a pure ACE inhibitor. Thus, inhibition of both ACE and NEP with omapatrilat could be more cardioprotective than ACE inhibition alone. PMID- 11046098 TI - An alpha(1A)/alpha(1L)-adrenoceptor mediates contraction of canine subcutaneous resistance arteries. AB - To determine the characteristics of the alpha(1)-adrenoceptor subtypes involved in adrenergic regulation of peripheral vascular resistance, contraction of canine subcutaneous resistance arteries was studied using wire myographs. The potencies of agonists and antagonists, chosen for their ability to discriminate between alpha(1)-adrenoceptor subtypes, were assessed in the presence of cocaine (3 microM), corticosterone (30 microM), and propranolol (1 microM). The rank order of agonist potency (pEC(50) +/- S.E.) was (R)-A-61603 (7.88 +/- 0.1) > norepinephrine (6.41 +/- 0.1) > phenylephrine (5.83 +/- 0.1). The high sensitivity to (R)-A-61603 relative to phenylephrine is inconsistent with the presence of the alpha(1D)-adrenoceptor and most consistent with an alpha(1A) adrenoceptor response. This is supported by the low affinity for the alpha(1D) selective antagonist BMY 7378 (pK(B) 6.51 +/- 0.47). The low pA(2) values for prazosin (8.36) and HV723 (8.81), by definition, indicate the involvement of the putative alpha(1L)-adrenoceptor, a hypothesis supported by the pA(2) values for WB4101 (8.42) and 5-methyl-urapidil (8.08). Pre-exposure to 1 microM CEC had little effect, whereas 100 microM CEC reduced the maximum contraction but not the sensitivity to norepinephrine. This low sensitivity to CEC argues against the presence of the alpha(1B)-adrenoceptor. We conclude that, by current definitions, an alpha(1A)-/alpha(1L)-adrenoceptor causes contraction of these vessels. This does not support the concept that selectivity for the alpha(1A)-adrenoceptor is the basis for the effectiveness of some alpha-blockers in some tissues, such as prostate, but not in other tissues such as blood vessels. Rather, the generally low potency of alpha-blockers in some tissues may be due to a tissue-specific property of the receptors. PMID- 11046099 TI - Oral cocaine pharmacokinetics and pharmacodynamics in a cumulative-dose regimen: pharmacokinetic-pharmacodynamic modeling of concurrent operant and spontaneous behavior within an operant context. AB - Despite wide use of cumulative-dosing procedures to evaluate dose-response relations, limited attention has been paid to investigating drug concentration effect relations. We first characterized the pharmacokinetic (PK) parameters for i.v. (2 mg/kg) and oral cocaine (20 and 40 mg/kg) in rats. Cocaine's concentration-time profile for the escalating cumulative-dose regimen was simulated from PK parameters, dose size (1, 2, 7, 20, and 45 mg/kg by the oral route), and dosing interval (tau, 35 min) as well as validated from blood sampling at various time points. This concentration-time profile was integrated with pharmacodynamic (PD) profiles of differential reinforcement of low rate performance and spontaneous activity (large and small movements) under a differential reinforcement of low rate 45-s schedule. Effects on three behavioral measures were characterized by integrated PK-PD models using the sigmoid E(max) (for increases in shorter response rate or large movements) and inhibitory E(max) (for decreases in density of reinforcement) models. But for the intrinsic differences in baseline and efficacy values among the behavioral endpoints, one set of PD parameters (i.e., potency and Hill factors) predicted concentration effect relations for the three behavioral indices across all five doses. Concurrent monitoring of operant and spontaneous activity behavior within an operant context provides a novel behavioral paradigm to investigate drug effects on spontaneous activity under conditions where a behavioral contingency exists. Additionally, a cumulative-dosing procedure is efficient for determining the entire dose-response relation and provides an ideal mode to study phenomena such as sensitization or tolerance by varying dose size and/or tau. PMID- 11046100 TI - Endogenous bradykinin and the renin and pressor responses to furosemide in humans. AB - In humans, bradykinin contributes to the acute renin response after ACE inhibition. To further explore the role of endogenous bradykinin in human renin regulation, we determined the effect of HOE 140, a specific bradykinin B(2) receptor antagonist, on the renin response to 0.5 mg/kg i.v. furosemide in a randomized, single blind, crossover design study of 10 healthy, salt-replete volunteers. HOE 140 did not affect basal plasma renin activity, aldosterone, mean arterial pressure, or heart rate. Furosemide administration increased plasma renin activity from 1.0 +/- 0.2 to 4.5 +/- 1.2 ng of angiotensin I/ml/h and there was no effect of HOE 140 (from 1.1 +/- 0.2 to 3.9 +/- 0.8 ng of angiotensin I/ml/h). Similarly, there was no effect of HOE 140 on the diuretic response to furosemide. Mean arterial pressure increased in response to furosemide after HOE 140 (82 +/- 2 to 94 +/- 2 mm Hg), but not after vehicle (81 +/- 3 to 85 +/- 2 mm Hg), whereas heart rate was unchanged. In conclusion, activation of the B(2) receptor by endogenous bradykinin does not contribute to the renin response to acute furosemide treatment in humans. However, bradykinin may contribute to blood pressure regulation under conditions in which the renin-angiotensin system is stimulated. PMID- 11046101 TI - Tasosartan, enoltasosartan, and angiotensin II receptor blockade: the confounding role of protein binding. AB - Tasosartan is a long-acting angiotensin II (AngII) receptor blocker. Its long duration of action has been attributed to its active metabolite enoltasosartan. In this study we evaluated the relative contribution of tasosartan and enoltasosartan to the overall pharmacological effect of tasosartan. AngII receptor blockade effect of single doses of tasosartan (100 mg p.o. and 50 mg i.v) and enoltasosartan (2.5 mg i.v.) were compared in 12 healthy subjects in a randomized, double blind, three-period crossover study using two approaches: the in vivo blood pressure response to exogenous AngII and an ex vivo AngII radioreceptor assay. Tasosartan induced a rapid and sustained blockade of AngII subtype-1 (AT1) receptors. In vivo, tasosartan (p.o. or i.v.) blocked by 80% AT1 receptors 1 to 2 h after drug administration and still had a 40% effect at 32 h. In vitro, the blockade was estimated to be 90% at 2 h and 20% at 32 h. In contrast, the blockade induced by enoltasosartan was markedly delayed and hardly reached 60 to 70% despite the i.v. administration and high plasma levels. In vitro, the AT1 antagonistic effect of enoltasosartan was markedly influenced by the presence of plasma proteins, leading to a decrease in its affinity for the receptor and a slower receptor association rate. The early effect of tasosartan is due mainly to tasosartan itself with little if any contribution of enoltasosartan. The antagonistic effect of enoltasosartan appears later. The delayed in vivo blockade effect observed for enoltasosartan appears to be due to a high and tight protein binding and a slow dissociation process from the carrier. PMID- 11046102 TI - Identification of R146225 as a novel, orally active inhibitor of interleukin-5 biosynthesis. AB - Interleukin (IL)-5 regulates the growth, differentiation, and activation of eosinophils. When activated, eosinophils release an array of proinflammatory and cytotoxic products and act as prominent effector cells in the process of allergic inflammation. Depriving eosinophils of IL-5 may therefore represent a viable approach to treat allergic disorders. This study describes the identification of R146225, a novel six-substituted azauracil derivative, as a potent, orally active inhibitor of IL-5 biosynthesis, capable of reducing pulmonary eosinophilia in mice. In vitro, R146225 inhibited IL-5 protein formation by activated human whole blood (IC(50) = 34 nM), human peripheral blood mononuclear cells (IC(50) = 24 nM), and murine spleen cells (IC(50) = 6 nM). In contrast, the compound enhanced generation of interferon-gamma and had little or no inhibitory effect on the production of IL-2 and IL-4. Reverse transcription-polymerase chain reaction analysis of stimulated whole blood cells indicated R146225's ability to down regulate IL-5 mRNA expression. In vivo p.o. administration of R146225 (2.5 mg/kg) to mice before an i.v. anti-CD3 antibody challenge reduced IL-5 but enhanced interferon-gamma serum levels, without affecting IL-2 and IL-4 production. Analogous to the in vitro results, R146225 suppressed splenic IL-5 mRNA expression, while message levels of the other cytokines remained unchanged. Moreover, p.o. dosing of R146225 (0.6-2.5 mg/kg) dose dependently reduced the pulmonary accumulation of eosinophils induced in mice by an intranasal instillation of Cryptococcus neoformans. Based on these data, R146225 may be useful in the therapy of eosinophil-driven allergic conditions. PMID- 11046103 TI - A turnover model of irreversible inhibition of gastric acid secretion by omeprazole in the dog. AB - A turnover model for irreversible inhibition of gastric acid secretion by omeprazole in gastric fistula dogs was developed using data from studies with both short- and long-term measurement periods. In the short-term experiments, after stimulation of acid secretion with histamine, the dogs were infused i.v. with omeprazole and acid secretion was measured for 5 h. Dose and infusion times were varied to produce different concentration-time profiles and schedule dependence in the inhibitory effect of omeprazole was observed. In the long-term experiments, dogs were given single intraduodenal doses, which inhibited the acid secretion for several days. Combining the short-term and long-term data allowed the observation of a biphasic recovery of acid secretion that was described by the turnover model. Second order association rate constants (k(ome)) for the covalent binding of omeprazole to H(+),K(+)-ATPase were estimated to 11 and 3.0 l/micromol/h for the i.v. and intraduodenal experiments, respectively. The apparent turnover rate constant of the enzyme (k(out)) was 0.013 h(-1) and the corresponding half-life of inhibition of acid secretory capacity was 54 h. The potency, calculated as k(out) over k(ome), was 4.3 and 1.2 nM for the intraduodenal and i.v. doses, respectively. Allometric scaling of the model resulted in trustworthy predictions for observations previously done in humans. The model predicted a good correlation between maximal inhibitory effect and exposure (area under the plasma concentration curve). The time dependence in this relation was also predicted by the model. PMID- 11046104 TI - Glycoprotein IIb/IIIa receptor number and occupancy during chronic administration of an oral antagonist. AB - Long-term treatment with oral glycoprotein (GP)IIb/IIIa antagonists has failed to produce significant clinical benefit. We have examined the pharmacology of xemilofiban in the evaluation of oral xemilofiban in controlling thrombotic events (EXCITE) trial. The EXCITE trial was a multicenter study of xemilofiban in 7232 patients undergoing percutaneous coronary intervention. Thirty-two patients randomized to xemilofiban (10 or 20 mg three times daily) or placebo were followed for up to 6 months. GPIIb/IIIa receptor number and occupancy were quantified using two monoclonal antibodies mAb1 and mAb2. mAb1 was used to quantify receptor number. mAb2 recognizes an epitope that is lost due to a ligand induced conformational change in GPIIb/IIIa and is a marker of receptor occupancy. Platelet aggregation was performed by light transmission. In vitro, the active metabolite of xemilofiban (SC-54701) inhibited mAb2 binding (IC(50) of 0.5 +/- 0.1 x 10(-8) M) but not mAb1. In vivo, long-term therapy with xemilofiban did not alter GPIIb/IIIa receptor number. mAb2 binding was inhibited throughout the treatment period and recovered slowly after drug withdrawal. Maximum inhibition of ADP-induced aggregation occurred at 4 to 7 h after the first dose of study medication. However, inhibition of platelet aggregation was low (between 24 and 45%) before dosing on days 60 and 180. There was no significant rebound increase in platelet aggregation after drug withdrawal. Long-term xemilofiban therapy does not alter platelet GPIIb/IIIa receptor number. Inhibition of platelet aggregation was poor at the end of each dosing interval and this may explain the failure of xemilofiban to alter clinical events. PMID- 11046105 TI - Efficacy of LGD1069 (Targretin), a retinoid X receptor-selective ligand, for treatment of uterine leiomyoma. AB - The conventional treatment of uterine leiomyomas, or fibroids, with gonadotropin releasing hormone (GnRH) agonists is often associated with serious side effects, necessitating short-term, palliative use of this therapy. Therefore, we examined a retinoid X receptor (RXR)-selective ligand, LGD1069, as a possible treatment for leiomyoma. LGD1069 has demonstrated efficacy as a chemopreventive agent in the N-nitroso-N-methylurea (NMU)-induced rat mammary carcinoma model and is a therapeutic agent in several epithelial tumor models. Previous studies have shown that it has both antitumor effects and antiestrogenic activity in the rat uterus, suggesting the potential utility of this agent for treatment of hormonally dependent uterine fibroids. The expression of retinoid receptors in tumors and cell lines derived from leiomyomas arising in the Eker rat was confirmed by Northern analysis. After treatment for 4 months with LGD1069, the number of grossly observable tumors was substantially reduced although the total incidence of tumors, including microscopic lesions, remained unaffected, suggesting an effect of the compound on tumor growth kinetics rather than on tumor initiation. Analysis of terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) staining and determination of 5-bromo-2-deoxyuridine (BrdU) incorporation indicated that the reduction in grossly observable tumors that occurred in treated animals was mediated by a significant increase in the level of apoptosis rather than a decrease in cell proliferation. These results suggest that LGD1069 may be an effective therapeutic agent for uterine leiomyoma that may inhibit tumor growth and, consequently, alleviate the symptoms associated with this disease. PMID- 11046106 TI - Cross-chimeric analysis of selectivity of secretin and VPAC(1) receptor activation. AB - Agonist action at receptors is highly specific, affected by the structure of both ligand and receptor. Chimeric constructs of structurally related receptors and/or ligands that have biological differences provide an opportunity to correlate a specific structural domain with function. In this work, we have used a cross chimeric approach to explore the structural basis for rat secretin and vasoactive intestinal polypeptide action at their closely related secretin and VPAC(1) receptors, belonging to class II of the G protein-coupled receptor superfamily. Multiple domains of both ligands and receptors contributed toward their selectivity, with differing combinations of such domains able to support high potency interactions. The amino-terminal 15 residues of secretin were most critical for potent stimulation of secretin receptors, whereas either the amino- or carboxyl-terminal halves of vasoactive intestinal polypeptide, when complemented by Lys(15), provided potent stimulation of the VPAC(1) receptor. The amino terminus of the VPAC(1) receptor was most critical for potent response to vasoactive intestinal polypeptide, whereas the amino terminus of the secretin receptor was important, but not adequate, requiring the complementation of an extracellular loop domain for potent response to secretin. Differences in the distribution of these determinants within these receptors provided an opportunity to produce a more "universal" receptor that contained the first extracellular loop of the secretin receptor and the remainder of the VPAC(1) receptor. This cross-chimeric approach should be applied to other members of this receptor family to test the emerging themes and to expand these insights as broadly as possible. PMID- 11046107 TI - Down-regulation of benzodiazepine binding to alpha 5 subunit-containing gamma aminobutyric Acid(A) receptors in tolerant rat brain indicates particular involvement of the hippocampal CA1 region. AB - Chronic benzodiazepine treatment can produce tolerance and changes in gamma aminobutyric acid (GABA)(A) receptors. To study the effect of treatment on a selected population of receptors, assays were performed using [(3)H]RY-80, which is selective for GABA(A) receptors with an alpha 5 subunit. Rats were given a flurazepam treatment known to produce tolerance and down-regulation of benzodiazepine binding, or a diazepam treatment shown to produce tolerance but not receptor down-regulation. Quantitative receptor autoradiography using sagittal brain sections bound with [(3)H]RY-80 showed binding in areas known to express alpha 5 mRNA. Brains from flurazepam-treated rats showed significantly decreased 1 nM [(3)H]RY-80 binding in hippocampal formation (e.g., 32% decrease in CA1) and superior colliculus, but not other areas. Using 5 nM [(3)H]RY-80 showed similar decreases in hippocampus. A corresponding 29% decrease in B(max) but no change in K(d) was found with a filtration binding assay using hippocampal homogenates. Down-regulation of [(3)H]RY-80 binding had returned to control by 2 days after withdrawing flurazepam treatment. The magnitude of down-regulation of [(3)H]RY-80 binding suggested that GABA(A) receptors with an alpha 5 subunit may play a prominent role in the adaptive responses associated with benzodiazepine tolerance. Chronic diazepam treatment also resulted in decreased [(3)H]RY-80 binding. However, the regional selectivity was even more pronounced than in flurazepam-treated rats, and only the hippocampal CA1 region showed decreased binding (27%). This localized down-regulation persisted for several days after the end of diazepam treatment. These data indicate that synapses in the hippocampal CA1 region are particularly involved in the adaptive response to chronic benzodiazepine treatments. PMID- 11046108 TI - Losartan improves recovery of contraction and inhibits transient inward current in a cellular model of cardiac ischemia and reperfusion. AB - Losartan, a selective angiotensin II (AII) type I receptor antagonist, may protect against myocardial stunning and arrhythmia in ischemia and reperfusion. To examine the cellular basis for these protective actions, we studied effects of losartan and AII on contractile and electrical activity of ventricular myocytes exposed to simulated ischemia and reperfusion. Ionic currents were measured with voltage-clamp techniques and contractions were measured with a video edge detector. After 10 min of superfusion with Tyrode's solution at 37 degrees C, cells were exposed to simulated ischemia (hypoxia, acidosis, hyperkalemia, hypercapnia, lactate accumulation, and substrate deprivation) for 30 min followed by 25 min of reperfusion with normal Tyrode's solution. During ischemia, drug treated cells were exposed to either 0.1 microM AII, 10 microM losartan, or both simultaneously. In reperfusion, contractions were depressed to 42% of preischemic levels in untreated cells. Losartan treatment significantly improved contractile recovery to 84% (P <. 05) of preischemic levels. AII-treated cells showed contractile recovery similar to untreated cells (40%), whereas cells treated with losartan plus AII recovered to 101% of preischemic levels. Cells exposed to losartan or losartan plus AII also exhibited reduced incidence of transient inward current (I(TI)) (20%, P <.05; 36%) relative to untreated cells (60%). However, I(TI) incidence was not altered by treatment with AII alone (57%). Treatment with exogenous agonist did not potentiate contractile depression or I(TI) incidence, and losartan exerted protective effects in the presence and absence of AII. Thus, losartan may have effects that are independent of AII receptor blockade. PMID- 11046109 TI - Putative link between transcriptional regulation of IgM expression by 2,3,7,8 tetrachlorodibenzo-p-dioxin and the aryl hydrocarbon receptor/dioxin-responsive enhancer signaling pathway. AB - The B-cell, a major cellular component of humoral immunity, has been identified as a sensitive target of 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD). The actual molecular mechanism responsible for the immunotoxic effects produced by TCDD is unclear; however, many of the biological effects produced by TCDD are thought to be mediated by the aryl hydrocarbon receptor (AhR). Using the CH12.LX B-cell line, the present studies show that inhibition of mu gene expression and IgM protein secretion by polychlorinated dibenzo-p-dioxin congeners follow a structure-activity relationship for AhR binding. Furthermore, these effects may be mediated by the two dioxin-responsive enhancer (DRE)-like sites that were identified within the Ig heavy chain 3'alpha-enhancer. Electrophoretic mobility shift assay-Western analysis demonstrated TCDD-induced binding of the AhR nuclear complex to both DRE-like sites as well as TCDD-induced binding of several nuclear factor-kappa B/Rel proteins to a kappa B site, which overlaps one of the DRE-like sites. Interestingly, kappa B binding in the AhR-deficient BCL-1 B-cells was also induced by TCDD, demonstrating an AhR-independent effect of TCDD on kappa B binding. Taken together, these results support an AhR/DRE-mediated mechanism for TCDD-induced inhibition of IgM expression. PMID- 11046110 TI - Polarized efflux of mono- and diacid metabolites of ME3229, an ester-type prodrug of a glycoprotein IIb/IIIa receptor antagonist, in rat small intestine. AB - ME3229 is an ester-type prodrug of a glycoprotein IIb/IIIa receptor antagonist ME3277. In our previous study, it was shown that only a small part of the drug taken up into the enterocytes reached the mesenteric vein, mainly due to transporter-mediated efflux of its hydrolyzed metabolites formed in the cells. To characterize the efflux transport system for the metabolites, the transport of the diacid metabolite ME3277 and the monoacid metabolites PM-10 and PM-11 were studied. ME3277 and PM-10 were preferentially transported in the serosal-to mucosal direction across the rat small intestine in the presence of glucose. Permeability of ME3277 across monolayer of Caco-2 cells with P-glycoprotein (P gp) and indomethacin-sensitive efflux pump expression did not show any directionality and verapamil, an inhibitor of P-gp, and indomethacin did not affect the permeability of ME3277 across rat intestinal tissue. Directional transport was not site specific and was observed in the Eisai hyperbilirubinemic rat whose canalicular multispecific organic anion transporter/multidrug resistance-associated protein (cMOAT/MRP2) is hereditarily defective as well as in normal rats. The efflux transport of ME3277 was inhibited by 1-naphthol, 1 choloro-2,4-dinitrobenzene, and sulfobromophthalein, and efflux of ME3277 and monoacid metabolites from intestinal tissue preloaded with ME3229 fell in the presence of 1-naphthol and sulfobromophthalein. These results demonstrate that mono- and diacid metabolites of ME3229 were pumped out into the gut lumen by an energy-dependent transport system located on the mucosal membrane of intestinal tissue and distinct from either P-gp, indomethacin-sensitive efflux pump or canalicular multispecific organic anion transporter/MRP2. An inhibition study suggested that this unknown transporter has a substrate specificity similar to that of MRP transporter families. PMID- 11046111 TI - Sensitivity of myelomonocytic leukemia cells to arsenite-induced cell cycle disruption, apoptosis, and enhanced differentiation is dependent on the inter relationship between arsenic concentration, duration of treatment, and cell cycle phase. AB - Arsenite treatment has been found to induce clinical remission in patients with acute promyelocytic leukemia. Although the potential therapeutic value of arsenite may lie in triggering apoptosis, it has not been established that cytotoxicity is the sole mechanism of action. We have used a myelomonocytic leukemia cell line (U937) to characterize the concentration-dependent effects of arsenite on cell growth, viability, apoptosis, and differentiation. Arsenite has multiple effects on U937 cells. Low concentrations of arsenite (i.e., < or = 1 microM) potentiate vitamin-D(3)-induced differentiation. Two markers of monocyte differentiation, Mac-1 expression and nitroblue tetrazolium reduction, are increased in arsenite-exposed, D(3)-costimulated cells. Concentrations of arsenite >10 microM rapidly induce the death of cells irrespective of cell cycle phase. Intermediate concentrations of arsenite (i.e., 5 to 10 microM) are cytostatic initially. Cell cycle analysis using elutriated, synchronous cell populations revealed that intermediate concentrations of arsenite delay both G(1) and G(2) transit. G(2) cells appear to be most sensitive to arsenite, in that transit through G(2)/M is more delayed than transit through G(1), and apoptosis is induced in these cells as they emerge from an aberrant G(2)/M. Arsenite induced apoptosis was caspase-3 dependent. Arsenite-mediated cytotoxicity was reduced in the presence of the broad caspase inhibitor Z-Val-Ala-DL-Asp fluoromethylketone; however, caspase inhibition did not reverse arsenite-induced cytostasis. Thus, arsenite has multiple effects on U937 cells that are dependent on concentration and cell cycle phase. Specifically, cell cycle transit and differentiation are more sensitive to arsenite than is the induction of apoptosis. PMID- 11046112 TI - Semiphysiological model for the time course of leukocytes after varying schedules of 5-fluorouracil in rats. AB - Models of leukopenia after chemotherapy are mainly empirical. To increase the derived models' potential of mechanistic understanding and extrapolation, more physiologically based models are being developed. To date, presented models cannot characterize the often-observed rebound of leukocytes. Therefore, a model able to describe the transient decrease and rebound in leukocytes was developed. Three different dosing regimens of 5-fluorouracil were given to rats. One group received a single dose of 127 mg/kg. The other two groups received two and three injections of 63 mg/kg and 49 mg/kg, respectively, with a 2-day interval. Leukocyte counts were followed for 23 to 25 days after the first dose. Plasma concentrations were determined by high-performance liquid chromatography. Population pharmacokinetic and pharmacodynamic models were developed using NONMEM. 5-Fluorouracil showed one-compartment disposition with capacity-limited elimination. The 49-mg/kg dose injected on three occasions produced the lowest leukocyte count (28% of baseline) and the most prominent rebound of the schedules, despite the fact that the fractionated regimens produced only 52 to 56% of the area under the concentration-time curve from time 0 to infinity in the single-dose group. The final semiphysiological model included two 5-fluorouracil sensitive and two -insensitive transit compartments as well as a compartment of circulating leukocytes. Second order rate constants from the transit compartments and a negative feedback from the circulating leukocytes to the input of the first sensitive compartment characterized the pronounced changes in leukocyte counts. A posterior predictive check as well as predictions into a new data set showed that our model could well predict the schedule-dependent leukopenic effects of 5 fluorouracil. PMID- 11046113 TI - The in vitro ethanol sensitivity of hippocampal synaptic gamma-aminobutyric acid(A) responses differs in lines of mice and rats genetically selected for behavioral sensitivity or insensitivity to ethanol. AB - Previous work has demonstrated that in the hippocampal CA1 region of Sprague Dawley rats, there are ethanol-sensitive and ethanol-insensitive populations of GABAergic synapses on pyramidal neurons. The present experiments characterized the ethanol sensitivity of these pathways in lines of rats and mice genetically selected for sensitivity or insensitivity to the behavioral effects of ethanol. In ethanol-sensitive inbred long sleep mice, GABA(A) IPSCs induced by stimulation of proximal (probably somatic) synapses were enhanced by 80 mM ethanol, whereas the distal (i.e., dendritic) pathway was unaffected. Thus, the relative sensitivity of these pathways (proximal > distal) is the same in both Sprague Dawley rats and in inbred long sleep mice. However, in the ethanol-insensitive inbred short sleep mice, neither proximal nor distal IPSCs were affected by 80 mM ethanol. The ethanol sensitivity of the proximal pathway was also examined in replicate lines of rats selected for either high ethanol sensitivity or low ethanol sensitivity. GABA(A) IPSCs in the high ethanol sensitivity lines were significantly enhanced by 80 mM ethanol, whereas IPSCs in the low ethanol sensitivity lines were unaffected. Thus, IPSCs evoked via the proximal pathway were enhanced by ethanol in all the sensitive mouse and rat lines, and unaffected in all the insensitive lines. These experiments demonstrate that GABA(A) synapses in brain differ in their sensitivity to enhancement by ethanol, and the sensitivity to such enhancement is under the control of genes that can be selected for using classical genetic selective breeding based on a behavioral phenotype. PMID- 11046114 TI - Chronic intragastric infusion of ethanol-containing diets induces CYP3A9 while decreasing CYP3A2 in male rats. AB - The CYP3A subfamily is the most abundant of the human hepatic cytochrome P450 enzymes. They mediate the biotransformation of many drugs, including a number of psychotropic, cardiac, analgesic, hormonal, immunosuppressant, antineoplastic, and antihistaminic agents. We studied diet/ethanol interactions using total enteral nutrition in adult male Sprague-Dawley rats with diets containing 16% protein, ethanol (13 g/kg), corn oil (fat; 25-45%), and carbohydrate (CHO; 1 21%). Using this model, chronic ethanol feeding decreased CYP3A activity (testosterone 6 beta-hydroxylation) and apoprotein levels (Western blot) (P <.05) and these effects were independent of the dietary CHO/fat ratio. The CYP3A2 mRNA levels decreased (P <.05) in the rats fed ethanol-containing diets by 73 to 83% compared with rats fed control diets, regardless of the CHO/fat ratio. In contrast, ethanol induced CYP3A9 mRNA levels (P <.05) and this effect was greater (P <.05) in the high-CHO/low-fat group (11.3-fold) than in the low-CHO/high-fat group (2.6-fold). Purified recombinant rat P450 3A9 had a chlorzoxazone 6 hydroxylase activity with a turnover number 1.3 nmol/min/nmol of P450. These results indicate that 1) ethanol differentially affects the expression of CYP3A gene family and this regulation appears to be modulated by dietary CHO/fat ratio; 2) the decrease in testosterone 6 beta-hydroxylase activity and CYP3A apoprotein levels are most likely due to the ethanol-induced decrease in CYP3A2 mRNA levels; and 3) CYP3A9 is induced by ethanol and is a low-affinity, high-K(m) chlorzoxazone hydroxylase. PMID- 11046115 TI - Beneficial insulin-sensitizing and vascular effects of S15261 in the insulin resistant JCR:LA-cp rat. AB - S15261, a compound developed for the oral treatment of type II diabetes, is cleaved by esterases to the fragments Y415 and S15511. The aim was to define the insulin-sensitizing effects of S15261, the cleavage products, and troglitazone and metformin in the JCR:LA-cp rat, an animal model of the obesity/insulin resistance syndrome that exhibits an associated vasculopathy and cardiovascular disease. Treatment of the animals from 8 to 12 weeks of age with S15261 or S15511 resulted in reductions in food intake and body weights, whereas Y415 had no effect. Troglitazone caused a small increase in food intake (P <.05). Treatment with S15261 or S15511 decreased plasma insulin levels in fed rats and prevented the postprandial peak in insulin levels in a meal tolerance test. Y415 had no effect on insulin levels. Troglitazone halved the insulin response to the test meal, but metformin gave no improvement. S15261 decreased the expression of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase and stimulated the expression of acetyl-CoA carboxylase and acyl-CoA synthase. S15261 also reduced the expression of carnitine palmitoyltransferase I and hydroxymethyl-glutaryl-CoA synthase. S15261, but not troglitazone, reduced the exaggerated contractile response of mesenteric resistance vessels to norepinephrine, and increased the maximal nitric oxide-mediated relaxation. S15261, through S15511, increased insulin sensitivity, decreased insulin levels, and reduced the vasculopathy of the JCR:LA-cp rat. S15261 may thus offer effective treatment for the insulin resistance syndrome and its associated vascular complications. PMID- 11046116 TI - Tuberoinfundibular peptide (7-39) [TIP(7-39)], a novel, selective, high-affinity antagonist for the parathyroid hormone-1 receptor with no detectable agonist activity. AB - The parathyroid hormone (PTH)-1 receptor mediates the pathophysiological effects of PTH in hyperparathyroidism and PTH-related protein (PTHrP) in humoral hypercalcemia of malignancy. A PTH1 receptor antagonist may be of therapeutic utility in these disorders. We recently identified a novel antagonist, tuberoinfundibular peptide (7-39) [TIP(7-39)], derived from the likely endogenous ligand for the PTH2 receptor TIP39. In this study its in vitro profile is evaluated and compared with that of [D-Trp(12),Tyr(34)]bPTH(7-34) and PTHrP(7 34), representing the two previously known structural classes of PTH1 receptor antagonists. TIP(7-39) binds with higher affinity (6.2 nM) to the PTH1 receptor than [D-Trp(12),Tyr(34)]bPTH(7-34) (45 nM) and PTHrP(7-34) (65 nM) and displays a 5.5-fold greater PTH1/PTH2 receptor selectivity. TIP(7-39) does not stimulate cAMP accumulation via the PTH1 receptor [in a sensitive assay that detects the activity of the weak partial agonist [Nle(8,18),Tyr(34)]bPTH(3-34)] and does not increase intracellular calcium. Schild analysis for TIP(7-39) was consistent with purely competitive antagonism of PTH(1-34)'s stimulation of cAMP accumulation (slope = 0.99 +/- 0.24). The pK(B) for TIP(7-39) (7.1 +/- 0.3) was higher than that for [D-Trp(12),Tyr(34)]bPTH(7-34) (6.5 +/- 0.0) and PTHrP(7-34) (6.0 +/- 0.1). Binding of (125)I-TIP(7-39) to the PTH1 receptor could be measured (K(D) = 1.3 +/- 0.1 nM, B(max) = 1.3 +/- 0.1 pmol/mg), whereas binding of (125)I [Nle(8,18),D-Trp(12),Tyr(34)]bPTH(7-34) could not be detected. Kinetic analysis indicated that (125)I-TIP(7-39) dissociates much more slowly (t(1/2) = 14 min) than [D-Trp(12),Tyr(34)]bPTH(7-34) (13 s) and PTHrP(7-34) (9 s). The novel antagonist TIP(7-39) therefore displays a more favorable in vitro pharmacological profile than antagonists derived from PTH and PTHrP and may be useful for demonstrating the utility of PTH1 receptor antagonism in the treatment of hypercalcemia. PMID- 11046117 TI - Characterization of mibefradil block of the human heart delayed rectifier hKv1.5. AB - The goal of this study was to analyze the effects of mibefradil on a human cardiac K(+) channel (hKv1.5) stably expressed in Chinese hamster ovary cells using the whole-cell configuration of the patch-clamp technique. Mibefradil inhibited in a concentration-dependent manner the hKv1.5 current with a K(D) value of 0.78 +/- 0.05 microM and a Hill coefficient of 0.97 +/- 0.06. Block induced by mibefradil was voltage dependent, consistent with a value of electrical distance of 0.13. The apparent association (k) and dissociation (l) rate constants measured at +50 mV were found to be 7.3 +/- 0.5 x 10(6) M(-1).s( 1) and 4.3 +/- 0.1 s(-1), respectively. Block increased rapidly between -20 and +10 mV, coincident with channel opening and suggested an open channel block mechanism, which was confirmed by a slower deactivation time course resulting in a "crossover" phenomenon when tail currents recorded under control conditions and in the presence of mibefradil were superimposed. Shifts toward negative potentials of the maximum conductance and the activation curve were observed, confirming the voltage dependence of block. Mibefradil induced a significant use dependent block when trains of depolarization at frequencies between 0.02 and 2 Hz were applied. In the presence of mibefradil, recovery of inactivation was faster than under control conditions, suggesting that mibefradil might compete with the inactivation gate of hKv1.5. These results indicate that mibefradil blocks hKv1.5 channels in a concentration-, voltage-, time- and use-dependent manner and the concentrations needed to observe these effects are in the therapeutic range. PMID- 11046118 TI - Potent and reversible effects of ATI-2001 on atrial and atrioventricular nodal electrophysiological properties in guinea pig isolated perfused heart. AB - We recently demonstrated that the short-acting analog of amiodarone, ATI-2001, caused favorable effects in guinea pig ventricular myocardium on electrophysiological substrates underlying tachyarrhythmia initiation, perpetuation, and termination. Here, the acute effects of 1.0 microM ATI-2001 and 1.0 microM amiodarone (90-min infusion followed by 90-min washout period) on atrial and atrioventricular (AV) nodal electrophysiological properties were studied in guinea pig isolated hearts. Neither ATI-2001 nor amiodarone significantly prolonged atrial conduction time. Compared with amiodarone, ATI 2001 caused significantly more rapid and greater prolongation of atrial monophasic action potential duration at 90% repolarization (maximal change 21.4 +/- 3.7 versus 19.0 +/- 4.0 ms) and atrial effective refractory period (ERP, 27.8 +/- 6.1 versus 9.2 +/- 2.3 ms). Shortening of the atrial cycle length from 250 to 200 ms did not significantly alter drug-induced changes in atrial repolarization and refractoriness. ATI-2001 prolonged the atrium-to-His bundle interval (22.1 +/ 2.6 versus 8.8 +/- 2.3 ms), His bundle-to-ventricle interval (2.8 +/- 0.4 versus 0.9 +/- 0.3 ms), AV nodal ERP (72.5 +/- 7.3 versus 31.4 +/- 4.1 ms), and Wenckebach cycle length (69.6 +/- 5.2 versus 35.8 +/- 4.1 ms) significantly more than did amiodarone. Unlike amiodarone, the effects of ATI-2001 were markedly reversed upon discontinuation of drug infusion. Given these data, ATI-2001 should not only be useful for terminating ongoing and preventing reoccurrence of atrial tachyarrhythmias but also to treat supraventricular tachycardias involving the AV node and to control ventricular rate during atrial tachyarrhythmias. Whether the observed differences in the pharmacokinetic properties render ATI-2001 superior to amiodarone in acute tachyarrhythmia management and less likely to accumulate into tissues during chronic therapy remains to be established. PMID- 11046119 TI - Nitric oxide stimulatory and endothelial protective effects of idoxifene, a selective estrogen receptor modulator, in the splanchnic artery of the ovariectomized rat. AB - Estrogen is known to stimulate endothelial nitric oxide production and attenuate endothelial dysfunction after ischemia and reperfusion. However, estrogen therapy increases the risk of breast and endometrial cancer. The present study was designed to determine whether idoxifene, a selective estrogen receptor modulator without adverse effects on reproductive organs, may stimulate nitric oxide release and protect endothelial function. In U-46619 precontracted superior mesenteric arterial (SMA) segments isolated from ovariectomized rats, idoxifene and 17 beta-estradiol resulted in a comparable dose-dependent vasorelaxation (maximal relaxation: 75.3 +/- 4.9 and 71 +/- 4.7%, respectively). Treatment of the rings with N(omega)-nitro-L-arginine methyl ester completely blocked idoxifene- and 17 beta-estradiol-induced vasorelaxation. In vitro incubation of SMA rings with TNF alpha significantly reduced vasorelaxation to an endothelium dependent vasodilator, acetylcholine (maximal relaxation: 73 +/- 3.7 versus 95 +/ 2.9% pre-TNF alpha, P <.01). Idoxifene, but surprisingly not 17 beta-estradiol, prevented TNF alpha-induced endothelial dysfunction (maximal relaxation: 86 +/- 2.6% in idoxifene-treated rings and 77 +/- 5.1% in 17beta-estrogen-treated rings). In vivo ischemia and reperfusion resulted in significant endothelial dysfunction as evidenced by decreased vasorelaxation to acetylcholine (maximal relaxation: 48 +/- 5.5 versus 92 +/- 3.9% in normal SMA rings), but a normal relaxation response to an endothelium-independent vasodilator, acidified NaNO(2) (95 +/- 3.2%). Treatment with idoxifene at either 1 or 2 mg/kg/day, or 17beta estrogen at 1 mg/kg/day for 4 days significantly preserved endothelial function (P <.01 versus vehicle). Taken together, these results demonstrate that idoxifene is an endothelium-dependent vasodilator and exerts significant endothelial protective effects against TNF alpha- and ischemia-reperfusion-induced endothelial injury. These results suggest that selective estrogen receptor modulators have therapeutic potential in diseases where endothelial dysfunction plays an important role. PMID- 11046120 TI - Scaling drug biotransformation data from cDNA-expressed cytochrome P-450 to human liver: a comparison of relative activity factors and human liver abundance in studies of mirtazapine metabolism. AB - The present study represents a comparison of three approaches to transform recombinant cytochrome P-450 (rCYP) enzyme kinetic data to human liver activity using mirtazapine (MIR) biotransformation as a model. MIR metabolite rCYP formation rates were corrected using I) relative activity factors (RAFs) determined on site, II) RAFs based on activity data provided by the rCYP manufacturer, and III) immunologically determined human liver abundance of CYP isoforms reported in the literature. For 2.5, 25, and 250 microM MIR, predictions of 1) the relative contribution of CYP isoforms to a particular reaction, 2) absolute metabolite formation rates, 3) the relative contribution of each pathway to net MIR biotransformation, and 4) the relative contribution of CYP isoforms to net MIR biotransformation were generated, and the results were compared with data obtained with human liver microsomes (HLM). We found that RAFs determined on site most accurately predict the results observed in HLM. Estimations based on liver abundance systematically underestimated CYP1A2 and overestimated CYP3A and CYP2C9 contributions to MIR metabolism and, therefore, seem less suitable to predict CYP isoform involvement in a particular reaction. Normalized RAFs calculated from the manufacturer activity data fell within the range of RAFs determined on site and lead to similar results for CYP isoform contribution to metabolic reactions and to net MIR biotransformation. Considering the time and resource-intensive step of RAF determination, manufacturer RAFs are an alternative to RAFs determined on site for the transformation of rCYP enzyme kinetic data; both of them provide more accurate estimations than immunologically determined human liver CYP isoform content. PMID- 11046121 TI - Cyclooxygenase-2-mediated angiogenesis in carrageenin-induced granulation tissue in rats. AB - The possible participation of cyclooxygenase (COX)-2 in angiogenesis in granulation tissue was analyzed using an air pouch-type carrageenin-induced inflammation model in rats. Injection of carrageenin solution into an air pouch induced gradual increases in the pouch fluid volume and granulation tissue weight as well as angiogenesis in granulation tissue. NS-398 (10-100 microg) inhibited all of these parameters in a dose-dependent manner. NS-398 (100 microg), indomethacin (100 microg), and dexamethasone (10 microg) markedly reduced prostaglandin (PG) E(2) levels in the pouch fluid at day 6. NS-398 and indomethacin did not affect protein levels of COX-1 and COX-2 but dexamethasone significantly reduced the level of COX-2 in granulation tissue at day 6. Protein levels of vascular endothelial growth factor (VEGF) in granulation tissue and in the pouch fluid were higher at day 6 than at day 3, and the levels were decreased by treatment with NS-398 (10-100 microg) in a dose-dependent manner. The inhibitory effects of NS-398 (100 microg) were almost the same as those of indomethacin (100 microg). Dexamethasone (10 microg) also reduced VEGF protein levels in granulation tissue at day 6. To clarify the role of PGE(2) in VEGF production, minced granulation tissue obtained 3 days after carrageenin injection from the indomethacin-treated rats was incubated in the presence of various concentrations of PGE(2). It was shown that VEGF mRNA and protein levels in the minced granulation tissue were increased by PGE(2) in a concentration-dependent manner. These findings suggest that COX-2-derived PGE(2) plays a significant role in angiogenesis in the carrageenin-induced granulation tissue through VEGF formation. PMID- 11046122 TI - Functional evidence of a constitutively active population of alpha(1D) adrenoceptors in rat aorta. AB - After depletion of intracellular calcium stores sensitive to noradrenaline, a spontaneous increase in the resting tone (IRT) when incubated in Ca(2+) containing solution was observed in isolated rat aorta, but not in tail artery. This IRT does not depend on agonist activation of alpha(1)-adrenoceptors but it is inhibited by prazosin. A close relationship was found between the inhibitory potencies of prazosin (pIC(50) = 9.833), BMY 7378 (pIC(50) = 8.924), and 5 methylurapidil (pIC(50) = 7.883) against IRT and their affinities for cloned alpha(1D)-adrenoceptors. Chloroethylclonidine (100 micromol. l(-1)) did not inhibit the IRT. After depletion of internal calcium stores by noradrenaline in absence of the agonist, loading in Ca(2+)-containing solution also brings about an increase in the inositol phosphate (IP) levels in rat aorta (not seen in tail artery) that is inhibited by prazosin (1 micromol. l(-1)), BMY 7378 (10 micromol. l(-1)), and 5-methylurapidil (10 micromol. l(-1)), thus confirming the results obtained in contractile studies. Chloroethylclonidine (100 micromol. l(-1)) did not inhibit this IP accumulation. The fact that the IRT and the IP accumulation related to it can be selectively inhibited by different alpha(1)-adrenoceptor antagonists suggests the existence of a population of alpha(1D)-adrenoceptors that show constitutive activity in rat aorta, not in tail artery. PMID- 11046123 TI - Inhibition of endothelial cell activation by nitric oxide donors. AB - Because nitric oxide (NO) inhibits the expression of endothelial leukocyte adhesion molecules, NO-generating compounds have major therapeutic potential for use outside their classical indications. We report on the in vitro potential antiatherogenicity of two novel cysteine-containing NO donors, SP/W 3672, a fast spontaneous NO releaser, and its prodrug SP/W 5186, which liberates NO after bioactivation. The ability of these two compounds to inhibit monocyte adhesion and surface expression of endothelial adhesion molecules was evaluated and compared with that of other NO donors. SP/W 5186 and SP/W 3672 inhibited the adhesion of U937 monocytes to cultured human endothelial cells more potently than S-nitrosoglutathione (GSNO) or spermine NONOate, whereas nitroglycerin and isosorbide dinitrate were ineffective at comparable concentrations. A similar rank order of potency was found for the inhibition of expression of the adhesion molecules vascular cell adhesion molecule-1, intercellular adhesion molecule-1, and E-selectin as well as for major histocompatibility complex class II antigen expression. Estimated IC(50) values for vascular cell adhesion molecule-1 were >400 microM for SP/W 4744 (control for SP/W 3672 lacking the cysteine moiety), 200 microM for GSNO and spermine NONOate, 80 microM for SP/W 3672, and 50 microM for SP/W 5186. Moreover, SP/W 5186 inhibited VCAM-1 mRNA levels more potently than GSNO. This effect was likely to be transcriptional because mRNA degradation was not affected. In conclusion, SP/W 3672 and SP/W 5186 are novel potent inhibitors of endothelial activation, and this effect appears to relate to their ability to liberate NO for prolonged periods of time, either spontaneously or after conversion to active hydrolytic products. PMID- 11046124 TI - Nitric oxide and NK(1)-tachykinin receptors in cyclophosphamide-induced cystitis, in rats. AB - The present study was conducted to investigate the role of NK(1) receptors and of nitric oxide (NO) on the pathogenesis of cyclophosphamide-induced cystitis, in rats. This bladder toxicity was characterized by marked increases in protein plasma extravasation, urothelial damage, edema, white blood cell infiltrates, and vascular congestion. These changes were associated with appearance of Ca(2+) independent NO-synthase (NOS) activity [characteristic of inducible NOS (iNOS)] in the bladder and with increases in urinary NO metabolites. GR205171, a selective NK(1) antagonist (10-20 mg/kg, i.p.) reduced cyclophosphamide-induced increases in protein plasma extravasation and in the urinary excretion of NO metabolites. N(G)-Nitro-L-arginine (L-NNA) (10 mg/kg, i.p.), a NOS inhibitor, reduced basal and cyclophosphamide-induced increases in NO metabolites and protected against cyclophosphamide-induced protein plasma extravasation. GR205171 had no effect, whereas L-NNA reduced basal NO metabolite excretion. Combined treatment with the NK(1) antagonist and the NO-synthesis inhibitor produced comparable reduction in protein plasma extravasation than that achieved with each drug given separately. Combined drug treatment ameliorated cyclophosphamideinduced urothelial damage, and the extent of edema, vascular congestion, and white blood cell infiltrates in the bladder. In summary, NK(1) receptors and iNOS play a role in NO formation and on cyclophosphamide-induced cystitis. Activation of NK(1) receptors mainly acts through the formation of NO. It is proposed that cyclophosphamide and/or its metabolites would stimulate primary afferent capsaicin-sensitive fibers in the bladder, releasing neuropeptides, which would activate NK(1) receptors. However, additional mechanisms are involved, because neither the NK(1) receptor antagonist nor the NO synthesis inhibitor, either alone or in combination, were able to completely prevent the toxicity. PMID- 11046125 TI - Right heart failure impairs hepatic elimination of p-nitrophenol without inducing changes in content or latency of hepatic UDP-glucuronosyltransferases. AB - Congestive heart failure has been shown to affect oxidative drug metabolism, however, there has been little study of its effects on drug conjugation. Using the isolated perfused livers from rats with right ventricular failure (RVF) due to pulmonary artery constriction, we studied the effects of RVF on hepatic elimination of p-nitrophenol (PNP) under controlled flow and oxygen delivery conditions. Hepatic clearance of the drug was found to be significantly impaired in RVF as compared with the sham group (0.80 +/- 0.23 versus 1.28 +/- 0.26 ml/min/g of liver). The impairment of PNP clearance in RVF occurred in parallel with significant reduction in metabolic formation clearance of p-nitrophenyl-beta D-glucuronide; the major metabolite of PNP (0.51 +/- 0.12 versus 1.03 +/- 0.26 ml/min/g of liver). The intrinsic drug-glucuronidation capacity of livers was evaluated by measuring the microsomal content and activity of the UDP glucuronosyltransferase(s) (UDP-GT) toward p-nitrophenol. There was no significant difference between sham and the RVF groups in either the content or the activity of the UDP-GT. The latency of the UDP-GT enzymes in microsomes was measured and was found to be similar between the two groups. The results of this study show that RVF impairs hepatic elimination of PNP and that this appears to be independent of changes in hepatic perfusion and oxygenation or alterations in hepatic content, activity, and latency of the UDP-GT. PMID- 11046126 TI - Evaluation of route of input on the hepatic disposition of diazepam. AB - Diazepam, a drug of high intrinsic clearance, was studied in the in situ rat liver dually perfused with Krebs-bicarbonate buffer containing human serum albumin (HSA; 0-1%) and unlabeled diazepam (1 mg/l) under constant hepatic arterial (3 ml/min) and portal venous (PV; 12 ml/min) flow rates. Events after a unit impulse (using [(14)C]diazepam) and at steady state (using unlabeled diazepam) were evaluated. In the absence of HSA the fractional effluent recovery (F) after hepatic arterial infusion (0.046 +/- 0.013) was about twice that after PV infusion (0.019 +/- 0.006). With HSA present, regardless of input route, F increased as unbound diazepam fraction in perfusate decreased (e.g., for PV, F = 0.58 +/- 0.05 and 0.69 +/- 0.02 for unbound diazepam fraction values of 0.18 +/- 0.01 and 0.037 +/- 0.01 at 0.25% and 1% HSA). The effluent [(14)C]diazepam profile was also dependent upon HSA. On decreasing HSA from 1 to 0.25% the early sharp peak (at 12-20 s) was replaced by a flatter unimodal profile with a later peak (at 60-80 s). Comparison of estimated effective permeability-surface area product to perfusate flow ratios (4.4 for 1% HSA and 21 for 0.25% HSA) indicated a shift from a perfusion rate-limited uptake with 0.25% HSA to one intermediate between permeability and perfusion at 1% HSA. Recognizing that orally absorbed drug enters the liver only via PV and i.v. drug via both vascular routes, this study emphasizes the difference in hepatic extraction of compounds depending on route of input, and the role that alteration in perfusate binding has on hepatic drug disposition. PMID- 11046127 TI - 5-hydroxylation of omeprazole by human liver microsomal fractions from Chinese populations related to CYP2C19 gene dose and individual ethnicity. AB - It has been previously reported that omeprazole (OP) oxidation is mediated by CYP2C19 and CYP3A4 in human livers. In this study, we assessed their relative contributions with human liver microsomal fractions from Chinese populations that were genotyped by CYP2C19 and recruited from two ethnic groups, Han and Zhuang. The kinetics of 5-hydroxyomeprazole (5-OH-OP) formation was best described by the two-enzyme and single-enzyme Michaelis-Menten equations for liver microsomes from CYP2C19 extensive (EMs) and poor metabolizers, respectively. At a low substrate concentration that may be encountered in vivo, the monoclonal antibody to CYP2C8/9/19 strongly inhibited 5-OH-OP formation in EM microsomes, whereas troleandomycin (TAO) eliminated most of the formation at a high substrate concentration. In poor metabolizer microsomes, either TAO or anti-CYP3A4 could alone abolish 5-OH-OP formation. Furthermore, there were differences between homozygous and heterozygous EMs in the percentage of inhibition by TAO and the antibodies. At the low substrate concentration, OP 5-hydroxyaltion was correlated well with S-mephenytoin 4'-hydroxylation and CYP2C19 contents in liver microsomes of 34 Chinese individuals. Moreover, in these individuals, obviously genetic and somewhat ethnic differences in OP 5-hydroxylation were observed between different CYP2C19 genotypes (wt/wt > wt/m1 > m1/m1) and between Han and Zhuang (Han > Zhuang), respectively. The results indicate that CYP2C19 is a high-affinity enzyme for OP 5-hydroxylation by liver microsomes from Chinese individuals and that its contribution is CYP2C19 gene dependent and ethnically related. Similar studies indicate that OP sulfoxidation is mediated mainly by CYP3A4 and independent of CYP2C19 genotype status. PMID- 11046128 TI - Selection of alternative 5' splice sites: role of U1 snRNP and models for the antagonistic effects of SF2/ASF and hnRNP A1. AB - The first component known to recognize and discriminate among potential 5' splice sites (5'SSs) in pre-mRNA is the U1 snRNP. However, the relative levels of U1 snRNP binding to alternative 5'SSs do not necessarily determine the splicing outcome. Strikingly, SF2/ASF, one of the essential SR protein-splicing factors, causes a dose-dependent shift in splicing to a downstream (intron-proximal) site, and yet it increases U1 snRNP binding at upstream and downstream sites simultaneously. We show here that hnRNP A1, which shifts splicing towards an upstream 5'SS, causes reduced U1 snRNP binding at both sites. Nonetheless, the importance of U1 snRNP binding is shown by proportionality between the level of U1 snRNP binding to the downstream site and its use in splicing. With purified components, hnRNP A1 reduces U1 snRNP binding to 5'SSs by binding cooperatively and indiscriminately to the pre-mRNA. Mutations in hnRNP A1 and SF2/ASF show that the opposite effects of the proteins on 5'SS choice are correlated with their effects on U1 snRNP binding. Cross-linking experiments show that SF2/ASF and hnRNP A1 compete to bind pre-mRNA, and we conclude that this competition is the basis of their functional antagonism; SF2/ASF enhances U1 snRNP binding at all 5'SSs, the rise in simultaneous occupancy causing a shift in splicing towards the downstream site, whereas hnRNP A1 interferes with U1 snRNP binding such that 5'SS occupancy is lower and the affinities of U1 snRNP for the individual sites determine the site of splicing. PMID- 11046129 TI - Developmentally regulated rpd3p homolog specific to the transcriptionally active macronucleus of vegetative Tetrahymena thermophila. AB - A clear relationship exists between histone acetylation and transcriptional output, the balance of which is conferred by opposing histone acetyltransferases (HATs) and histone deacetylases (HDACs). To explore the role of HDAC activity in determining the transcriptional competency of chromatin, we have exploited the biological features of Tetrahymena as a model. Each vegetative cell contains two nuclei: a somatic, transcriptionally active macronucleus containing hyperacetylated chromatin and a transcriptionally silent, germ line micronucleus containing hypoacetylated histones. Using a PCR-based strategy, a deacetylase gene (named THD1) encoding a homolog of the yeast HDAC Rpd3p was cloned. Thd1p deacetylates all four core histones in vitro. It resides exclusively in the macronucleus during vegetative growth and is asymmetrically distributed to developing new macronuclei early in their differentiation during the sexual pathway. Together, these data are most consistent with a potential role for Thd1p in transcriptional regulation and suggest that histone deacetylation may be important for the differentiation of micronuclei into macronuclei during development. PMID- 11046130 TI - Cloning and characterization of two novel thyroid hormone receptor beta isoforms. AB - Thyroid hormone (T(3)) activates nuclear receptor transcription factors, encoded by the TRalpha (NR1A1) and TRbeta (NR1A2) genes, to regulate target gene expression. Several TR isoforms exist, and studies of null mice have identified some unique functions for individual TR variants, although considerable redundancy occurs, raising questions about the specificity of T(3) action. Thus, it is not known how diverse T(3) actions are regulated in target tissues that express multiple receptor variants. I have identified two novel TRbeta isoforms that are expressed widely and result from alternative mRNA splicing. TRbeta3 is a 44.6-kDa protein that contains an unique 23-amino-acid N terminus and acts as a functional receptor. TRDeltabeta3 is a 32.8-kDa protein that lacks a DNA binding domain but retains ligand binding activity and is a potent dominant-negative antagonist. The relative concentrations of beta3 and Deltabeta3 mRNAs vary between tissues and with changes in thyroid status, indicating that alternative splicing is tissue specific and T(3) regulated. These data provide novel insights into the mechanisms of T(3) action and define a new level of specificity that may regulate thyroid status in tissue. PMID- 11046131 TI - Functional interaction between Ssu72 and the Rpb2 subunit of RNA polymerase II in Saccharomyces cerevisiae. AB - SSU72 is an essential gene encoding a phylogenetically conserved protein of unknown function that interacts with the general transcription factor TFIIB. A recessive ssu72-1 allele was identified as a synthetic enhancer of a TFIIB (sua7 1) defect, resulting in a heat-sensitive (Ts(-)) phenotype and a dramatic downstream shift in transcription start site selection. Here we describe a new allele, ssu72-2, that confers a Ts(-) phenotype in a SUA7 wild-type background. In an effort to further define Ssu72, we isolated suppressors of the ssu72-2 mutation. One suppressor is allelic to RPB2, the gene encoding the second-largest subunit of RNA polymerase II (RNAP II). Sequence analysis of the rpb2-100 suppressor defined a cysteine replacement of the phylogenetically invariant arginine residue at position 512 (R512C), located within homology block D of Rpb2. The ssu72-2 and rpb2-100 mutations adversely affected noninduced gene expression, with no apparent effects on activated transcription in vivo. Although isolated as a suppressor of the ssu72-2 Ts(-) defect, rpb2-100 enhanced the transcriptional defects associated with ssu72-2. The Ssu72 protein interacts directly with purified RNAP II in a coimmunoprecipitation assay, suggesting that the genetic interactions between ssu72-2 and rpb2-100 are a consequence of physical interactions. These results define Ssu72 as a highly conserved factor that physically and functionally interacts with the RNAP II core machinery during transcription initiation. PMID- 11046132 TI - Platelet-derived growth factor receptor association with Na(+)/H(+) exchanger regulatory factor potentiates receptor activity. AB - Platelet-derived growth factor (PDGF) is a potent mitogen for many cell types. The PDGF receptor (PDGFR) is a receptor tyrosine kinase that mediates the mitogenic effects of PDGF by binding to and/or phosphorylating a variety of intracellular signaling proteins upon PDGF-induced receptor dimerization. We show here that the Na(+)/H(+) exchanger regulatory factor (NHERF; also known as EBP50), a protein not previously known to interact with the PDGFR, binds to the PDGFR carboxyl terminus (PDGFR-CT) with high affinity via a PDZ (PSD-95/Dlg/Z0-1 homology) domain-mediated interaction and potentiates PDGFR autophosphorylation and extracellular signal-regulated kinase (ERK) activation in cells. A point mutated version of the PDGFR, with the terminal leucine changed to alanine (L1106A), cannot bind NHERF in vitro and is markedly impaired relative to the wild-type receptor with regard to PDGF-induced autophosphorylation and activation of ERK in cells. NHERF potentiation of PDGFR signaling depends on the capacity of NHERF to oligomerize. NHERF oligomerizes in vitro when bound with PDGFR-CT, and a truncated version of the first NHERF PDZ domain that can bind PDGFR-CT but which does not oligomerize reduces PDGFR tyrosine kinase activity when transiently overexpressed in cells. PDGFR activity in cells can also be regulated in a NHERF dependent fashion by stimulation of the beta(2)-adrenergic receptor, a known cellular binding partner for NHERF. These findings reveal that NHERF can directly bind to the PDGFR and potentiate PDGFR activity, thus elucidating both a novel mechanism by which PDGFR activity can be regulated and a new cellular role for the PDZ domain-containing adapter protein NHERF. PMID- 11046133 TI - Sok2 regulates yeast pseudohyphal differentiation via a transcription factor cascade that regulates cell-cell adhesion. AB - In response to nitrogen limitation, Saccharomyces cerevisiae undergoes a dimorphic transition to filamentous pseudohyphal growth. In previous studies, the transcription factor Sok2 was found to negatively regulate pseudohyphal differentiation. By genome array and Northern analysis, we found that genes encoding the transcription factors Phd1, Ash1, and Swi5 were all induced in sok2/sok2 hyperfilamentous mutants. In accord with previous studies of others, Swi5 was required for ASH1 expression. Phd1 and Ash1 regulated expression of the cell surface protein Flo11, which is required for filamentous growth, and were largely required for filamentation of sok2/sok2 mutant strains. These findings reveal that a complex transcription factor cascade regulates filamentation. These findings also reveal a novel dual role for the transcription factor Swi5 in regulating filamentous growth. Finally, these studies illustrate how mother daughter cell adhesion can be accomplished by two distinct mechanisms: one involving Flo11 and the other involving regulation of the endochitinase Cts1 and the endoglucanase Egt2 by Swi5. PMID- 11046134 TI - The absence of Msh2 alters abelson virus pre-B-cell transformation by influencing p53 mutation. AB - Defects in DNA mismatch repair predispose cells to the development of several types of malignant disease. The absence of Msh2 or Mlh1, two key molecules that mediate mismatch repair in eukaryotic cells, increases the frequency of mutation and also alters the response of some cells to apoptosis and cell cycle arrest. To understand the way these changes contribute to cancer predisposition, we examined the effects of defective mismatch repair on the multistep process of pre-B-cell transformation by Abelson murine leukemia virus. In this model, primary transformants undergo a prolonged apoptotic crisis followed by the emergence of fully transformed cell lines. The latter event is correlated to a loss of function of the p53 tumor suppressor protein and down-modulation of the p53 regulatory protein p19Arf. Analyses of primary transformants from Msh2 null mice and their wild-type littermates revealed that both types of cells undergo crisis. However, primary transformants from Msh2 null animals recover with accelerated kinetics, a phenomenon that is strongly correlated to the appearance of cells that have lost p53 function. Analysis of the kinetics with which p53 function is lost revealed that this change provides the dominant stimulus for emergence from crisis. Therefore, the absence of mismatch repair alters the molecular mechanisms involved in transformation by affecting a gene that controls apoptosis and cell cycle progression, rather than by affecting these processes directly. PMID- 11046136 TI - Developmentally regulated excision of a 28-base-pair sequence from the Paramecium genome requires flanking DNA. AB - The micronuclear DNA of Paramecium tetraurelia is estimated to contain over 50,000 short DNA elements that are precisely removed during the formation of the transcriptionally active macronucleus. Each internal eliminated sequence (IES) is bounded by 5'-TA-3' dinucleotide repeats, a feature common to some classes of DNA transposons. We have developed an in vivo assay to analyze these highly efficient and precise DNA excision events. The microinjection of a cloned IES into mating cells results in accurately spliced products, and the transformed cells maintain the injected DNA as extrachromosomal molecules. A series of deletions flanking one side of a 28-bp IES were constructed and analyzed with the in vivo assay. Whereas 72 bp of DNA flanking the eliminated region is sufficient for excision, lengths of 31 and 18 bp result in reduced excision and removal of all wild-type sequences adjacent to the TA results in complete failure of excision. In contrast, nucleotide mutations within the middle of the 28-bp IES do not prevent excision. The results are consistent with a functional role for perfect inverted repeats flanking the IES. PMID- 11046135 TI - ERK5 is a novel type of mitogen-activated protein kinase containing a transcriptional activation domain. AB - Previous studies have shown that upregulation of the orphan steroid receptor Nur77 is required for the apoptosis of immature T cells in response to antigen receptor signals. Transcriptional upregulation of Nur77 in response to antigen receptor signaling involves two binding sites for the MEF2 family of transcription factors located in the Nur77 promoter. Calcium signals greatly increase the activity of MEF2D in T cells via a posttranslational mechanism. The mitogen-activated protein (MAP) kinase ERK5 was isolated in a yeast two-hybrid screen using the MADS-MEF2 domain of MEF2D as bait. ERK5 resembles the other MAP kinase family members in its N-terminal half, but it also contains a 400-amino acid C-terminal domain of previously uncharacterized function. We report here that the C-terminal region of ERK5 contains a MEF2-interacting domain and, surprisingly, also a potent transcriptional activation domain. These domains are both required for coactivation of MEF2D by ERK5. The MEF2-ERK5 interaction was found to be activation dependent in vivo and inhibitable in vitro by the calcium sensitive MEF2 repressor Cabin 1. The transcriptional activation domain of ERK5 is required for maximal MEF2 activity in response to calcium flux in T cells, and it can activate the endogenous Nur77 gene when constitutively recruited to the Nur77 promoter via MEF2 sites. These studies provide insights into a mechanism whereby MEF2 activity can respond to calcium signaling and suggest a novel, unexpected mechanism of MAP kinase function. PMID- 11046137 TI - Cdc13 cooperates with the yeast Ku proteins and Stn1 to regulate telomerase recruitment. AB - The Saccharomyces cerevisiae CDC13 protein binds single-strand telomeric DNA. Here we report the isolation of new mutant alleles of CDC13 that confer either abnormal telomere lengthening or telomere shortening. This deregulation not only depended on telomerase (Est2/TLC1) and Est1, a direct regulator of telomerase, but also on the yeast Ku proteins, yKu70/Hdf1 and yKu80/Hdf2, that have been previously implicated in DNA repair and telomere maintenance. Expression of a Cdc13-yKu70 fusion protein resulted in telomere elongation, similar to that produced by a Cdc13-Est1 fusion, thus suggesting that yKu70 might promote Cdc13 mediated telomerase recruitment. We also demonstrate that Stn1 is an inhibitor of telomerase recruitment by Cdc13, based both on STN1 overexpression and Cdc13-Stn1 fusion experiments. We propose that accurate regulation of telomerase recruitment by Cdc13 results from a coordinated balance between positive control by yKu70 and negative control by Stn1. Our results represent the first evidence of a direct control of the telomerase-loading function of Cdc13 by a double-strand telomeric DNA-binding complex. PMID- 11046138 TI - CD28 and the tyrosine kinase lck stimulate mitogen-activated protein kinase activity in T cells via inhibition of the small G protein Rap1. AB - Proliferation of T cells via activation of the T-cell receptor (TCR) requires concurrent engagement of accessory costimulatory molecules to achieve full activation. The best-studied costimulatory molecule, CD28, achieves these effects, in part, by augmenting signals from the TCR to the mitogen-activated protein (MAP) kinase cascade. We show here that TCR-mediated stimulation of MAP kinase extracellular-signal-regulated kinases (ERKs) is limited by activation of the Ras antagonist Rap1. CD28 increases ERK signaling by blocking Rap1 action. CD28 inhibits Rap1 activation because it selectively stimulates an extrinsic Rap1 GTPase activity. The ability of CD28 to stimulate Rap1 GTPase activity was dependent on the tyrosine kinase Lck. Our results suggest that CD28-mediated Rap1 GTPase-activating protein activation can help explain the augmentation of ERKs during CD28 costimulation. PMID- 11046139 TI - p53 binds selectively to the 5' untranslated region of cdk4, an RNA element necessary and sufficient for transforming growth factor beta- and p53-mediated translational inhibition of cdk4. AB - One consequence of transforming growth factor beta (TGF-beta) treatment is inhibition of Cdk4 synthesis, and this is dependent on p53. Here, we show that the 5' untranslated region (UTR) of the cdk4 mRNA is both necessary and sufficient for wild-type p53-dependent TGF-beta-regulated translational inhibition of cdk4. Wild-type p53 bound selectively to the 5' UTR of the cdk4 mRNA and inhibited translation of RNAs that contain this region. RNA binding and translational control are two genetically separable functions of p53, as are specific and nonspecific RNA binding. Moreover, transactivation-defective mutants of p53 retain the ability to regulate cdk4 translation. Our findings suggest that p53 functions as a regulator of translation in response to TGF-beta in vivo. PMID- 11046140 TI - Multiple homing pathways used by yeast mitochondrial group II introns. AB - The yeast mitochondrial DNA group II introns aI1 and aI2 are retroelements that insert site specifically into intronless alleles by a process called homing. Here, we used patterns of flanking marker coconversion in crosses with wild-type and mutant aI2 introns to distinguish three coexisting homing pathways: two that were reverse transcriptase (RT) dependent (retrohoming) and one that was RT independent. All three pathways are initiated by cleavage of the recipient DNA target site by the intron-encoded endonuclease, with the sense strand cleaved by partial or complete reverse splicing, and the antisense strand cleaved by the intron-encoded protein. The major retrohoming pathway in standard crosses leads to insertion of the intron with unidirectional coconversion of upstream exon sequences. This pattern of coconversion suggests that the major retrohoming pathway is initiated by target DNA-primed reverse transcription of the reverse spliced intron RNA and completed by double-strand break repair (DSBR) recombination with the donor allele. The RT-independent pathway leads to insertion of the intron with bidirectional coconversion and presumably occurs by a conventional DSBR recombination mechanism initiated by cleavage of the recipient DNA target site by the intron-encoded endonuclease, as for group I intron homing. Finally, some mutant DNA target sites shift up to 43% of retrohoming to another pathway not previously detected for aI2 in which there is no coconversion of flanking exon sequences. This new pathway presumably involves synthesis of a full-length cDNA copy of the inserted intron RNA, with completion by a repair process independent of homologous recombination, as found for the Lactococcus lactis Ll.LtrB intron. Our results show that group II intron mobility can occur by multiple pathways, the ratios of which depend on the characteristics of both the intron and the DNA target site. This remarkable flexibility enables group II introns to use different recombination and repair enzymes in different host cells. PMID- 11046141 TI - Uridylate addition and RNA ligation contribute to the specificity of kinetoplastid insertion RNA editing. AB - RNA editing in Trypanosoma brucei inserts and deletes uridylates (U's) in mitochondrial pre-mRNAs under the direction of guide RNAs (gRNAs). We report here the development of a novel in vitro precleaved editing assay and its use to study the gRNA specificity of the U addition and RNA ligation steps in insertion RNA editing. The 5' fragment of substrate RNA accumulated with the number of added U's specified by gRNA, and U addition products with more than the specified number of U's were rare. U addition up to the number specified occurred in the absence of ligation, but accumulation of U addition products was slowed. The 5' fragments with the correct number of added U's were preferentially ligated, apparently by adenylylated RNA ligase since exogenously added ATP was not required and since ligation was eliminated by treatment with pyrophosphate. gRNA specified U addition was apparent in the absence of ligation when the pre-mRNA immediately upstream of the editing site was single stranded and more so when it was base paired with gRNA. These results suggest that both the U addition and RNA ligation steps contributed to the precision of RNA editing. PMID- 11046142 TI - Multiple C-terminal lysine residues target p53 for ubiquitin-proteasome-mediated degradation. AB - In normal cells, p53 is maintained at a low level by ubiquitin-mediated proteolysis, but after genotoxic insult this process is inhibited and p53 levels rise dramatically. Ubiquitination of p53 requires the ubiquitin-activating enzyme Ubc5 as a ubiquitin conjugation enzyme and Mdm2, which acts as a ubiquitin protein ligase. In addition to the N-terminal region, which is required for interaction with Mdm2, the C-terminal domain of p53 modulates the susceptibility of p53 to Mdm2-mediated degradation. To analyze the role of the C-terminal domain in p53 ubiquitination, we have generated p53 molecules containing single and multiple lysine-to-arginine changes between residues 370 and 386. Although wild type (WT) and mutant molecules show similar subcellular distributions, the mutants display a higher transcriptional activity than WT p53. Simultaneous mutation of lysine residues 370, 372, 373, 381, 382, and 386 to arginine residues (6KR p53 mutant) generates a p53 molecule with potent transcriptional activity that is resistant to Mdm2-induced degradation and is refractory to Mdm2-mediated ubiquitination. In contrast to WT p53, transcriptional activity directed by the 6KR p53 mutant fails to be negatively regulated by Mdm2. Those differences are also manifest in HeLa cells which express the human papillomavirus E6 protein, suggesting that p53 C-terminal lysine residues are also implicated in E6-AP mediated ubiquitination. These data suggest that p53 C-terminal lysine residues are the main sites of ubiquitin ligation, which target p53 for proteasome mediated degradation. PMID- 11046143 TI - Nup2p, a yeast nucleoporin, functions in bidirectional transport of importin alpha. AB - Import of proteins containing a classical nuclear localization signal (NLS) into the nucleus is mediated by importin alpha and importin beta. Srp1p, the Saccharomyces cerevisiae homologue of importin alpha, returns from the nucleus in a complex with its export factor Cse1p and with Gsp1p (yeast Ran) in its GTP bound state. We studied the role of the nucleoporin Nup2p in the transport cycle of Srp1p. Cells lacking NUP2 show a specific defect in both NLS import and Srp1p export, indicating that Nup2p is required for efficient bidirectional transport of Srp1p across the nuclear pore complex (NPC). Nup2p is located at the nuclear side of the central gated channel of the NPC and provides a binding site for Srp1p via its amino-terminal domain. We show that Nup2p effectively releases the NLS protein from importin alpha-importin and beta and strongly binds to the importin heterodimer via Srp1p. Kap95p (importin beta) is released from this complex by a direct interaction with Gsp1p-GTP. These data suggest that besides Gsp1p, which disassembles the NLS-importin alpha-importin beta complex upon binding to Kap95p in the nucleus, Nup2p can also dissociate the import complex by binding to Srp1p. We also show data indicating that Nup1p, a relative of Nup2p, plays a similar role in termination of NLS import. Cse1p and Gsp1p-GTP release Srp1p from Nup2p, which suggests that the Srp1p export complex can be formed directly at the NPC. The changed distribution of Cse1p at the NPC in nup2 mutants also supports a role for Nup2p in Srp1p export from the nucleus. PMID- 11046144 TI - Ras-dependent regulation of c-Jun phosphorylation is mediated by the Ral guanine nucleotide exchange factor-Ral pathway. AB - The transcription factor c-Jun is critically involved in the regulation of proliferation and differentiation as well as cellular transformation induced by oncogenic Ras. The signal transduction pathways that couple Ras activation to c Jun phosphorylation are still partially elusive. Here we show that an activated version of the Ras effector Rlf, a guanine nucleotide exchange factor (GEF) of the small GTPase Ral, can induce the phosphorylation of serines 63 and 73 of c Jun. In addition, we show that growth factor-induced, Ras-mediated phosphorylation of c-Jun is abolished by inhibitory mutants of the RalGEF-Ral pathway. These results suggest that the RalGEF-Ral pathway plays a major role in Ras-dependent c-Jun phosphorylation. Ral-dependent regulation of c-Jun phosphorylation includes JNK, a still elusive JNKK, and possibly Src. PMID- 11046145 TI - Acetylation by PCAF enhances CIITA nuclear accumulation and transactivation of major histocompatibility complex class II genes. AB - The class II transactivator (CIITA), the master regulator of the tissue-specific and interferon gamma-inducible expression of major histocompatibility complex class II genes, synergizes with the histone acetylase coactivator CBP to activate gene transcription. Here we demonstrate that in addition to CBP, PCAF binds to CIITA both in vivo and in vitro and enhances CIITA-dependent transcriptional activation of class II promoters. Accordingly, E1A mutants defective for PCAF or CBP interaction show reduced ability in suppressing CIITA activity. Interestingly, CBP and PCAF acetylate CIITA at lysine residues within a nuclear localization signal. We show that CIITA is shuttling between the nucleus and cytoplasm. The shuttling behavior and activity of the protein are regulated by acetylation: overexpression of PCAF or inhibition of cellular deacetylases by trichostatin A increases the nuclear accumulation of CIITA in a manner determined by the presence of the acetylation target lysines. Furthermore, mutagenesis of the acetylated residues reduces the transactivation ability of CIITA. These results support a novel function for acetylation, i.e., to regulate gene expression by stimulating the nuclear accumulation of an activator. PMID- 11046146 TI - Thyroglobulin repression of thyroid transcription factor 1 (TTF-1) gene expression is mediated by decreased DNA binding of nuclear factor I proteins which control constitutive TTF-1 expression. AB - Follicular thyroglobulin (TG) selectively suppresses the expression of thyroid restricted transcription factors, thereby altering the expression of thyroid specific proteins. In this study, we investigated the molecular mechanism by which TG suppresses the prototypic thyroid-restricted transcription factor, thyroid transcription factor 1 (TTF-1), in rat FRTL-5 thyrocytes. We show that the region between bp -264 and -153 on the TTF-1 promoter contains two nuclear factor I (NFI) elements whose function is involved in TG-mediated suppression. Thus, NFI binding to these elements is critical for constitutive expression of TTF-1; TG decreases NFI binding to the NFI elements in association with TG repression. NFI is a family of transcription factors that is ubiquitously expressed and contributes to constitutive and cell-specific gene expression. In contrast to the contribution of NFI proteins to constitutive gene expression in other systems, we demonstrate that follicular TG transcriptionally represses all NFI RNAs (NFI-A, -B, -C, and -X) in association with decreased NFI binding and that the RNA levels decrease as early as 4 h after TG treatment. Although TG treatment for 48 h results in a decrease in NFI protein-DNA complexes measured in DNA mobility shift assays, NFI proteins are still detectable by Western analysis. We show, however, that the binding of all NFI proteins is redox regulated. Thus, diamide treatment of nuclear extracts strongly reduces the binding of NFI proteins, and the addition of higher concentrations of dithiothreitol to nuclear extracts from TG-treated cells restores NFI-DNA binding to levels in extracts from untreated cells. We conclude that NFI binding to two NFI elements, at bp 264 to -153, positively regulates TTF-1 expression and controls constitutive TTF 1 levels. TG mediates the repression of TTF-1 gene expression by decreasing NFI RNA and protein levels, as well as by altering the binding activity of NFI, which is redox controlled. PMID- 11046147 TI - The tyrosine phosphatase SHP-2 is required for sustained activation of extracellular signal-regulated kinase and epithelial morphogenesis downstream from the met receptor tyrosine kinase. AB - Epithelial morphogenesis is critical during development and wound healing, and alterations in this program contribute to neoplasia. Met, the hepatocyte growth factor (HGF) receptor, promotes a morphogenic program in epithelial cell lines in matrix cultures. Previous studies have identified Gab1, the major phosphorylated protein following Met activation, as important for the morphogenic response. Gab1 is a docking protein that couples the Met receptor with multiple signaling proteins, including phosphatidylinositol-3 kinase, phospholipase Cgamma, the adapter protein Crk, and the tyrosine specific phosphatase SHP-2. HGF induces sustained phosphorylation of Gab1 and sustained activation of extracellular signal-regulated kinase (Erk) in epithelial Madin-Darby canine kidney cells. In contrast, epidermal growth factor fails to promote a morphogenic program and induces transient Gab1 phosphorylation and Erk activation. To elucidate the Gab1 dependent signals required for epithelial morphogenesis, we undertook a structure function approach and demonstrate that association of Gab1 with the tyrosine phosphatase SHP-2 is required for sustained Erk activation and for epithelial morphogenesis downstream from the Met receptor. Epithelial cells expressing a Gab1 mutant protein unable to recruit SHP-2 elicit a transient activation of Erk in response to HGF. Moreover, SHP-2 catalytic activity is required, since the expression of a catalytically inactive SHP-2 mutant, C/S, abrogates sustained activation of Erk and epithelial morphogenesis by the Met receptor. These data identify SHP-2 as a positive modulator of Erk activity and epithelial morphogenesis downstream from the Met receptor. PMID- 11046148 TI - Latent membrane protein 2A of Epstein-Barr virus binds WW domain E3 protein ubiquitin ligases that ubiquitinate B-cell tyrosine kinases. AB - The latent membrane protein (LMP) 2A of Epstein-Barr virus (EBV) is implicated in the maintenance of viral latency and appears to function in part by inhibiting B cell receptor (BCR) signaling. The N-terminal cytoplasmic region of LMP2A has multiple tyrosine residues that upon phosphorylation bind the SH2 domains of the Syk tyrosine kinase and the Src family kinase Lyn. The LMP2A N-terminal region also has two conserved PPPPY motifs. Here we show that the PPPPY motifs of LMP2A bind multiple WW domains of E3 protein-ubiquitin ligases of the Nedd4 family, including AIP4 and KIAA0439, and demonstrate that AIP4 and KIAA0439 form physiological complexes with LMP2A in EBV-positive B cells. In addition to a C2 domain and four WW domains, these proteins have a C-terminal Hect catalytic domain implicated in the ubiquitination of target proteins. LMP2A enhances Lyn and Syk ubiquitination in vivo in a fashion that depends on the activity of Nedd4 family members and correlates with destabilization of the Lyn tyrosine kinase. These results suggest that LMP2A serves as a molecular scaffold to recruit both B cell tyrosine kinases and C2/WW/Hect domain E3 protein-ubiquitin ligases. This may promote Lyn and Syk ubiquitination in a fashion that contributes to a block in B-cell signaling. LMP2A may potentiate a normal mechanism by which Nedd4 family E3 enzymes regulate B-cell signaling. PMID- 11046149 TI - Characterization of dFMR1, a Drosophila melanogaster homolog of the fragile X mental retardation protein. AB - Fragile X syndrome is the most common inherited form of mental retardation. It is caused by loss of FMR1 gene activity due to either lack of expression or expression of a mutant form of the protein. In mammals, FMR1 is a member of a small protein family that consists of FMR1, FXR1, and FXR2. All three members bind RNA and contain sequence motifs that are commonly found in RNA-binding proteins, including two KH domains and an RGG box. The FMR1/FXR proteins also contain a 60S ribosomal subunit interaction domain and a protein-protein interaction domain which mediates homomer and heteromer formation with each family member. Nevertheless, the specific molecular functions of FMR1/FXR proteins are unknown. Here we report the cloning and characterization of a Drosophila melanogaster homolog of the mammalian FMR1/FXR gene family. This first invertebrate homolog, termed dfmr1, has a high degree of amino acid sequence identity/similarity with the defined functional domains of the FMR1/FXR proteins. The dfmr1 product binds RNA and is similar in subcellular localization and embryonic expression pattern to the mammalian FMR1/FXR proteins. Overexpression of dfmr1 driven by the UAS-GAL4 system leads to apoptotic cell loss in all adult Drosophila tissues examined. This phenotype is dependent on the activity of the KH domains. The ability to induce a dominant phenotype by overexpressing dfmr1 opens the possibility of using genetic approaches in Drosophila to identify the pathways in which the FMR1/FXR proteins function. PMID- 11046151 TI - DNA replication progresses on the periphery of nuclear aggregates formed by the BCL6 transcription factor. AB - The BCL6 proto-oncogene, frequently alterated in non-Hodgkin lymphoma, encodes a POZ/zinc finger protein that localizes into discrete nuclear subdomains. Upon prolonged BCL6 overexpression in cells bearing an inducible BCL6 allele (UTA-L cells), these subdomains apparently coincide with sites of DNA synthesis. Here, we explore the relationship between BCL6 and replication by both electron and confocal laser scanning microscopy. First, by electron microscope analyses, we found that endogenous BCL6 is associated with replication foci. Moreover, we show that a relatively low expression level of BCL6 reached after a brief induction in UTA-L cells is sufficient to observe its targeting to mid, late, and at least certain early replication foci visualized by a pulse-labeling with bromodeoxyuridine (BrdU). In addition, when UTA-L cells are simultaneously induced for BCL6 expression and exposed to BrdU for a few hours just after the release from a block in mitosis, a nuclear diffuse BCL6 staining indicates cells in G(1), while cells in S show a more punctate nuclear BCL6 distribution associated with replication foci. Finally, ultrastructural analyses in UTA-L cells exposed to BrdU for various times reveal that replication progresses just around, but not within, BCL6 subdomains. Thus, nascent DNA is localized near, but not colocalized with, BCL6 subdomains, suggesting that they play an architectural role influencing positioning and/or assembly of replication foci. Together with its previously function as transcription repressor recruiting a histone deacetylase complex, BCL6 may therefore contribute to link nuclear organization, replication, and chromatin-mediated regulation. PMID- 11046150 TI - Saccharomyces cerevisiae cdc42p GTPase is involved in preventing the recurrence of bud emergence during the cell cycle. AB - The Saccharomyces cerevisiae Cdc42p GTPase interacts with multiple regulators and downstream effectors through an approximately 25-amino-acid effector domain. Four effector domain mutations, Y32K, F37A, D38E, and Y40C, were introduced into Cdc42p and characterized for their effects on these interactions. Each mutant protein showed differential interactions with a number of downstream effectors and regulators and various levels of functionality. Specifically, Cdc42(D38E)p showed reduced interactions with the Cla4p p21-activated protein kinase and the Bem3p GTPase-activating protein and cdc42(D38E) was the only mutant allele able to complement the Deltacdc42 null mutant. However, the mutant protein was only partially functional, as indicated by a temperature-dependent multibudded phenotype seen in conjunction with defects in both septin ring localization and activation of the Swe1p-dependent morphogenetic checkpoint. Further analysis of this mutant suggested that the multiple buds emerged consecutively with a premature termination of bud enlargement preceding the appearance of the next bud. Cortical actin, the septin ring, Cla4p-green fluorescent protein (GFP), and GFP-Cdc24p all predominantly localized to one bud at a time per multibudded cell. These data suggest that Cdc42(D38E)p triggers a morphogenetic defect post-bud emergence, leading to cessation of bud growth and reorganization of the budding machinery to another random budding site, indicating that Cdc42p is involved in prevention of the initiation of supernumerary buds during the cell cycle. PMID- 11046152 TI - Cyclooxygenase 2 promotes cell survival by stimulation of dynein light chain expression and inhibition of neuronal nitric oxide synthase activity. AB - Cyclooxygenase 2 (COX-2) inhibits nerve growth factor (NGF) withdrawal apoptosis in differentiated PC12 cells. The inhibition of apoptosis by COX-2 was concomitant with prevention of caspase 3 activation. To understand how COX-2 prevents apoptosis, we used cDNA expression arrays to determine whether COX-2 regulates differential expression of apoptosis-related genes. The expression of dynein light chain (DLC) (also known as protein inhibitor of neuronal nitric oxide synthase [PIN]) was significantly stimulated in PC12 cells overexpressing COX-2. The COX-2-dependent stimulation of DLC expression was, at least in part, mediated by prostaglandin E(2). Overexpression of DLC also inhibited NGF withdrawal apoptosis in differentiated PC12 cells. Stimulation of DLC expression resulted in an increased association of DLC/PIN with neuronal nitric oxide synthase (nNOS), thereby reducing nNOS activity. Furthermore, nNOS expression and activity were significantly increased in differentiated PC12 cells after NGF withdrawal. This increased nNOS activity as well as increased nNOS dimer after NGF withdrawal were inhibited by COX-2 or DLC/PIN overexpression. An nNOS inhibitor or a membrane-permeable superoxide dismutase (SOD) mimetic protected differentiated PC12 cells from NGF withdrawal apoptosis. In contrast, NO donors induced apoptosis in differentiated PC12 cells and potentiated apoptosis induced by NGF withdrawal. The protective effects of COX-2 on apoptosis induced by NGF withdrawal were also overcome by NO donors. These findings suggest that COX-2 promotes cell survival by a mechanism linking increased expression of prosurvival genes coupled to inhibition of NO- and superoxide-mediated apoptosis. PMID- 11046153 TI - Distinct p300-responsive mechanisms promote caspase-dependent apoptosis by human T-cell lymphotropic virus type 1 Tax protein. AB - The dysregulation of cellular apoptosis pathways has emerged as a critical early event associated with the development of many types of human cancers. Numerous viral and cellular oncogenes, aside from their inherent transforming properties, are known to induce programmed cell death, consistent with the hypothesis that genetic defects are required to support tumor survival. Here, we report that nuclear expression of the CREB-binding protein (CBP)/p300-binding domain of the human T-cell lymphotropic virus type 1 (HTLV-1) transactivator, Tax, triggers an apoptotic death-inducing signal during short-term clonal analyses, as well as in transient cell death assays. Coexpression of the antiapoptotic factor Bcl-2 increased serum stimulation; incubation with the chemical caspase inhibitor z-Val Ala-DL-Asp fluoromethylketone antagonized Tax-induced cell death. The CBP/p300 binding defective Tax mutants K88A and V89A exhibited markedly reduced cytotoxic effects compared to the wild-type Tax protein. Importantly, nuclear expression of the minimal CBP/p300-binding peptide of Tax induced apoptosis in the absence of Tax-dependent transcriptional activities, while its K88A counterpart did not cause cell death. Further, Tax-mediated apoptosis was effectively prevented by ectopic expression of the p300 coactivator. We also report that activation of the NF-kappaB transcription pathway by Tax, under growth arrest conditions, results in apoptosis that occurs independent of direct Tax coactivator effects. Our results allude to a novel pivotal role for the transcriptional coactivator p300 in determining cell fate and raise the possibility that dysregulated coactivator usage may pose an early barrier to transformation that must be selectively overcome as a prerequisite for the initiation of neoplasia. PMID- 11046154 TI - The matrix protein of vesicular stomatitis virus inhibits nucleocytoplasmic transport when it is in the nucleus and associated with nuclear pore complexes. AB - The matrix (M) protein of vesicular stomatitis virus (VSV) is a potent inhibitor of bidirectional nuclear transport. Here we demonstrate that inhibition occurs when M protein is in the nucleus of Xenopus laevis oocytes and that M activity is readily reversed by a monoclonal antibody (alphaM). We identify a region of M protein, amino acids 51 to 59, that is required both for inhibition of transport and for efficient recognition by alphaM. When expressed in transfected HeLa cells, M protein colocalizes with nuclear pore complexes (NPCs) at the nuclear rim. Moreover, mutation of a single amino acid, methionine 51, eliminates both transport inhibition and targeting to NPCs. We propose that M protein inhibits bidirectional transport by interacting with a component of the NPC or an NPC associated factor that participates in nucleocytoplasmic transport. PMID- 11046155 TI - Chromatin association of human origin recognition complex, cdc6, and minichromosome maintenance proteins during the cell cycle: assembly of prereplication complexes in late mitosis. AB - Evidence obtained from studies with yeast and Xenopus indicate that the initiation of DNA replication is a multistep process. The origin recognition complex (ORC), Cdc6p, and minichromosome maintenance (MCM) proteins are required for establishing prereplication complexes, upon which initiation is triggered by the activation of cyclin-dependent kinases and the Dbf4p-dependent kinase Cdc7p. The identification of human homologues of these replication proteins allows investigation of S-phase regulation in mammalian cells. Using centrifugal elutriation of several human cell lines, we demonstrate that whereas human Orc2 (hOrc2p) and hMcm proteins are present throughout the cell cycle, hCdc6p levels vary, being very low in early G(1) and accumulating until cells enter mitosis. hCdc6p can be polyubiquitinated in vivo, and it is stabilized by proteasome inhibitors. Similar to the case for hOrc2p, a significant fraction of hCdc6p is present on chromatin throughout the cell cycle, whereas hMcm proteins alternate between soluble and chromatin-bound forms. Loading of hMcm proteins onto chromatin occurs in late mitosis concomitant with the destruction of cyclin B, indicating that the mitotic kinase activity inhibits prereplication complex formation in human cells. PMID- 11046156 TI - A family of LIM-only transcriptional coactivators: tissue-specific expression and selective activation of CREB and CREM. AB - Transcription factors of the CREB family control the expression of a large number of genes in response to various signaling pathways. Regulation mediated by members of the CREB family has been linked to various physiological functions. Classically, activation by CREB is known to occur upon phosphorylation at an essential regulatory site (Ser133 in CREB) and the subsequent interaction with the ubiquitous coactivator CREB-binding protein (CBP). However, the mechanism by which selectivity is achieved in the identification of target genes, as well as the routes adopted to ensure tissue-specific activation, remains unrecognized. We have recently described the first tissue-specific coactivator of CREB family transcription factors, ACT (activator of CREM in testis). ACT is a LIM-only protein which associates with CREM in male germ cells and provides an activation function which is independent of phosphorylation and CBP. Here we characterize a family of LIM-only proteins which share common structural organization with ACT. These are referred to as four-and-a-half-LIM-domain (FHL) proteins and display tissue-specific and developmentally regulated expression. FHL proteins display different degrees of intrinsic activation potential. They provide powerful activation function to both CREB and CREM when coexpressed either in yeast or in mammalian cells, specific combinations eliciting selective activation. Deletion analysis of the ACT protein shows that the activation function depends on specific arrangements of the LIM domains, which are essential for both transactivation and interaction properties. This study uncovers the existence of a family of tissue-specific coactivators that operate through novel, CBP independent routes to elicit transcriptional activation by CREB and CREM. The future identification of additional partners of FHL proteins is likely to reveal unappreciated aspects of tissue-specific transcriptional regulation. PMID- 11046158 TI - Editor's page : the 2000 RadioGraphics monograph issue: musculoskeletal imaging PMID- 11046159 TI - From the RSNA Refresher Courses. Radiological Society of North America. Adult chronic hip pain: radiographic evaluation. AB - Adult chronic hip pain can be difficult to attribute to a specific cause, both clinically and radiographically. Yet, there are often subtle radiographic signs that point to traumatic, infectious, arthritic, neoplastic, congenital, or other causes. Stress fractures appear as a lucent line surrounded by sclerosis or as subtle lucency or sclerosis. Subtle femoral neck angulation, trabecular angulation, or a subcapital impaction line indicates an insufficiency fracture. Apophyseal avulsion fractures appear as a thin, crescentic, ossific opacity when viewed in tangent and as a subtle, disk-shaped opacity when viewed en face. Effusion, cartilage loss, and cortical bone destruction are diagnostic of a septic hip. Transient osteoporosis manifests as osteoporosis and effusion. The earliest finding of avascular necrosis is relative sclerosis in the femoral head. Subtle osteophytes or erosive change is indicative of arthropathy. Osteoarthritis can manifest as early cyst formation, small osteophytes, or buttressing of the femoral neck or calcar. Rheumatoid arthritis may manifest as classic osteopenia, uniform cartilage loss, and erosive change. A disturbance of the trabecular pattern might suggest an early permeative pattern due to a tumor. Knowledge of common causes of chronic hip pain will allow the radiologist to seek out these radiographic findings. PMID- 11046160 TI - From the RSNA Refresher Courses. Radiological Society of North America. Evaluation of the marrow space in the adult hip. AB - The adult pelvis and hip contain extensive marrow space in which a variety of processes may occur. Evaluation of this space requires an understanding of normal maturation and recognition that the marrow of the pelvis (axial skeleton) and that of the proximal femurs (appendicular skeleton) contain variable amounts of red and yellow marrow. At magnetic resonance (MR) imaging, this variability yields patterns in normal marrow ranging from very uniform and homogeneous signal intensity to patchy and heterogeneous signal intensity. The marrow space serves as a reflection of patient health and may herald developing anemia with reconversion of inactive to active marrow. Pathologic processes to be considered include marrow edema related to trauma, tumors, or infection; marrow ischemia and infarction; marrow infiltration from primary or secondary neoplasms or from infection; or complete loss of normal myeloid tissue in the marrow space. Each process can be effectively studied with MR imaging. PMID- 11046157 TI - Cell signaling switches HOX-PBX complexes from repressors to activators of transcription mediated by histone deacetylases and histone acetyltransferases. AB - The Hoxb1 autoregulatory element comprises three HOX-PBX binding sites. Despite the presence of HOXB1 and PBX1, this enhancer fails to activate reporter gene expression in retinoic acid-treated P19 cell monolayers. Activation requires cell aggregation in addition to RA. This suggests that HOX-PBX complexes may repress transcription under some conditions. Consistent with this, multimerized HOX-PBX binding sites repress reporter gene expression in HEK293 cells. We provide a mechanistic basis for repressor function by demonstrating that a corepressor complex, including histone deacetylases (HDACs) 1 and 3, mSIN3B, and N-CoR/SMRT, interacts with PBX1A. We map a site of interaction with HDAC1 to the PBX1 N terminus and show that the PBX partner is required for repression by the HOX-PBX complex. Treatment with the deacetylase inhibitor trichostatin A not only relieves repression but also converts the HOX-PBX complex to a net activator of transcription. We show that this activation function is mediated by the recruitment of the coactivator CREB-binding protein by the HOX partner. Interestingly, HOX-PBX complexes are switched from transcriptional repressors to activators in response to protein kinase A signaling or cell aggregation. Together, our results suggest a model whereby the HOX-PBX complex can act as a repressor or activator of transcription via association with corepressors and coactivators. The model implies that cell signaling is a direct determinant of HOX-PBX function in the patterning of the animal embryo. PMID- 11046161 TI - From the RSNA Refresher Courses. Radiological Society of North America. Chronic adult hip pain: MR arthrography of the hip. AB - This article describes the technique for performance and interpretation of magnetic resonance arthrography of the hip. A description of normal anatomy of the hip is presented, and the appearance of the abnormal labrum is discussed. Labral detachments and tears are the most common clinically significant abnormalities to be identified. These abnormalities are recognized on the basis of the presence of contrast material at the acetabular-labral interface or within the substance of the labrum. Many varied appearances of the labrum have been identified within the asymptomatic population, and the correlation of these appearances is contrasted with those of the abnormal labrum in symptomatic patients. To date, it is difficult to draw conclusions regarding the significance of an absent labrum or of a sulcus at the acetabular-labral junction. Experience suggests that an absent labrum in a symptomatic individual is pathologic and that a sulcus at the anterosuperior acetabular-labral junction may be a normal variant. PMID- 11046162 TI - CT of unusual iliopsoas compartment lesions. AB - The authors reviewed the anatomy of the iliopsoas compartment and a spectrum of unusual lesions affecting structures in this compartment, with emphasis on the role of computed tomography (CT). Lesions in the iliopsoas compartment are caused by acute infection, tumor, or hemorrhage. The knowledge of detailed clinical data can help improve the diagnostic accuracy, particularly with regard to primary iliopsoas lesions. CT is useful for delineating the source of secondary iliopsoas lesions, guiding biopsy, and performing follow-up of treated lesions. Nonenhanced CT can help detect fresh hemorrhage, fat-containing tumor, and calcification, whereas contrast material enhanced CT optimizes imaging of infection, tumor, and aneurysm. PMID- 11046163 TI - CT and MR arthrography of the normal and pathologic anterosuperior labrum and labral-bicipital complex. AB - Interpretation of computed tomographic and magnetic resonance arthrograms of the shoulder is complicated by normal variants of the labrum and glenohumeral ligaments. A superior sublabral recess is located at the 12 o'clock position and represents a normal recess between the superior labrum and the cartilage of the glenoid cavity. A sublabral foramen is located at the 2 o'clock position and represents localized detachment of the labrum from the glenoid rim. Buford complex is characterized by absence of the anterosuperior labrum and cordlike thickening of the middle glenohumeral ligament. Imaging features of damage to the anterior labrum include absence or detachment of the labrum and an irregular frayed appearance. Superior labrum anterior-to-posterior (SLAP) lesions are classified as type I (tear confined to the superior labrum), type II (labrum and biceps tendon detached from the superior glenoid), type III (bucket handle tear of the superior labrum), or type IV (bucket handle tear of the superior labrum with lateral extension into the biceps tendon). Increased distance between the labrum and the glenoid, an irregular appearance of the labral margin, or lateral extension of the separation may suggest a SLAP lesion rather than a normal anatomic variant. However, differentiation between normal variants and pathologic conditions and between various types of SLAP lesions remains difficult. PMID- 11046164 TI - Three layers of the medial capsular and supporting structures of the knee: MR imaging-anatomic correlation. AB - The authors used a three-layer approach to correlate the appearance of the capsule and ligaments of the medial side of the knee on magnetic resonance (MR) images with corresponding anatomic slices. MR images of six fresh cadaveric specimens were obtained by using a proton-density-weighted fast spin-echo sequence with a 256 x 512 matrix. Specimens were frozen and sliced with a band saw into 3. 0-mm-thick sections that corresponded to the MR images. Three layers were depicted on both anatomic slices and MR images. Layer 1 consisted of the deep crural fascia; layer 2, the superficial portion of the medial collateral ligament (MCL); and layer 3, the capsule, the deep portion of the MCL, the meniscofemoral and meniscotibial extensions of the deep portion of the MCL, and the patellomeniscal ligament. Along the anterior aspect of the medial side of the knee, layer 1 was fused with layer 2; along the posterior aspect of the knee, layer 2 was fused with layer 3. PMID- 11046165 TI - Lateral stabilizing structures of the knee: functional anatomy and injuries assessed with MR imaging. AB - The lateral aspect of the knee is stabilized by a complex arrangement of ligaments, tendons, and muscles. These structures can be demonstrated with routine spin-echo magnetic resonance (MR) imaging sequences performed in the sagittal, coronal, and axial planes. Anterolateral stabilization is provided by the capsule and iliotibial tract. Posterolateral stabilization is provided by the arcuate ligament complex, which comprises the lateral collateral ligament; biceps femoris tendon; popliteus muscle and tendon; popliteal meniscal and popliteal fibular ligaments; oblique popliteal, arcuate, and fabellofibular ligaments; and lateral gastrocnemius muscle. Injuries to lateral knee structures are less common than injuries to medial knee structures but may be more disabling. Most lateral compartment injuries are associated with damage to the cruciate ligaments and medial knee structures. Moreover, such injuries are frequently overlooked at clinical examination. Structures of the anterolateral quadrant are the most frequently injured; posterolateral instability is considerably less common. Practically all tears of the lateral collateral ligament are associated with damage to posterolateral knee structures. Most injuries of the popliteus muscle and tendon are associated with damage to other knee structures. MR imaging can demonstrate these injuries. Familiarity with the musculotendinous anatomy of the knee will facilitate accurate diagnosis with MR imaging. PMID- 11046166 TI - Traumatic musculotendinous injuries of the knee: diagnosis with MR imaging. AB - Magnetic resonance (MR) imaging is the imaging modality of choice for evaluation of acute traumatic musculotendinous injuries of the knee. Three discrete categories of acute injuries to the musculotendinous unit can be defined: muscle contusion, myotendinous strain, and tendon avulsion. Among the quadriceps muscles, the rectus femoris is the most susceptible to injury at the myotendinous junction due to its superficial location, predominance of type II fibers, eccentric muscle action, and extension across two joints. Among the muscles of the pes anserinus, the sartorius is the most susceptible to strain injury due to its superficial location and biarticular course. The classic fusiform configuration of the semimembranosus along with a propensity for eccentric actions also make it prone to strain injury. MR imaging findings associated with rupture of the iliotibial tract include discontinuity and edema, which are best noted on coronal images. The same mechanism of injury that tears the arcuate ligament from its fibular insertion can also result in avulsion injury of the biceps femoris. The gastrocnemius muscle is prone to strain injury due to its action across two joints and its superficial location. Injuries of the muscle belly and myotendinous junction of the popliteus are far more common than tendinous injuries. PMID- 11046167 TI - Mechanism-based pattern approach to classification of complex injuries of the knee depicted at MR imaging. AB - Complex knee injuries are common, often resulting from multiple forces: varus, valgus, hyperextension, hyperflexion, internal rotation, external rotation, anterior or posterior translation, and axial load. Certain combinations of forces are known to cause specific injury patterns. After a review of the literature, the authors developed a mechanism-based classification system based on patterns of bone marrow edema and ligament injury for complex knee injuries depicted at magnetic resonance imaging. The classification system takes into account knee position and forces and recognition of patterns of bone injury and complementary soft-tissue injury. Ten mechanism-based injury patterns were recognized: (a) pure hyperextension, (b) hyperextension with varus, (c) hyperextension with valgus, (d) pure valgus, (e) pure varus, (f) flexion with valgus and external rotation, (g) flexion with varus and internal rotation, (h) flexion with posterior tibial translation, (i) patellar dislocation (flexion, valgus, and internal rotation of femur on fixed tibia), and (j) direct trauma. Recognition of these patterns may help assess the full extent of knee injury, particularly at the posterolateral and posteromedial corners of the knee. PMID- 11046168 TI - Bone contusion patterns of the knee at MR imaging: footprint of the mechanism of injury. AB - Bone marrow contusions are frequently identified at magnetic resonance imaging after an injury to the musculoskeletal system. These osseous injuries may result from a direct blow to the bone, from compressive forces of adjacent bones impacting one another, or from traction forces that occur during an avulsion injury. The distribution of bone marrow edema is like a footprint left behind at injury, providing valuable clues to the associated soft-tissue injuries. Five contusion patterns with associated soft-tissue injuries occur in the knee: pivot shift injury, dashboard injury, hyperextension injury, clip injury, and lateral patellar dislocation. The classic bone marrow edema pattern seen following the pivot shift injury involves the posterolateral tibial plateau and the midportion of the lateral femoral condyle. Edema occurs in the anterior aspect of the proximal tibia following the dashboard injury. Hyperextension results in the "kissing" contusion pattern involving the anterior aspect of the proximal tibia and distal femur. The clip injury results in a prominent area of edema involving the lateral femoral condyle and a smaller area of edema involving the medial femoral condyle. Finally, lateral patellar dislocation results in edema involving the inferomedial patella and anterior aspect of the lateral femoral condyle. In many instances, the mechanism of injury can be determined by studying the distribution of bone marrow edema, which then enables one to predict with accuracy the specific soft-tissue abnormalities that are likely to be present. PMID- 11046169 TI - From the RSNA Refresher Courses. Radiological Society of North America. MR imaging of the ankle and foot. AB - Magnetic resonance (MR) imaging has opened new horizons in the diagnosis and treatment of many musculoskeletal diseases of the ankle and foot. It demonstrates abnormalities in the bones and soft tissues before they become evident at other imaging modalities. The exquisite soft-tissue contrast resolution, noninvasive nature, and multiplanar capabilities of MR imaging make it especially valuable for the detection and assessment of a variety of soft-tissue disorders of the ligaments (eg, sprain), tendons (tendinosis, peritendinosis, tenosynovitis, entrapment, rupture, dislocation), and other soft-tissue structures (eg, anterolateral impingement syndrome, sinus tarsi syndrome, compressive neuropathies [eg, tarsal tunnel syndrome, Morton neuroma], synovial disorders). MR imaging has also been shown to be highly sensitive in the detection and staging of a number of musculoskeletal infections including cellulitis, soft tissue abscesses, and osteomyelitis. In addition, MR imaging is excellent for the early detection and assessment of a number of osseous abnormalities such as bone contusions, stress and insufficiency fractures, osteochondral fractures, osteonecrosis, and transient bone marrow edema. MR imaging is increasingly being recognized as the modality of choice for assessment of pathologic conditions of the ankle and foot. PMID- 11046170 TI - Plantar fasciitis and fascial rupture: MR imaging findings in 26 patients supplemented with anatomic data in cadavers. AB - Understanding of the normal anatomy of the plantar aponeurosis (PA) and familiarity with pathologic conditions are required for an accurate evaluation of the patient with subcalcaneal heel pain. In this study, we evaluated the diagnostic capabilities of magnetic resonance (MR) imaging in the assessment of the PA with close anatomic correlation. Herein, we describe the MR imaging features of plantar fasciitis and fascial rupture in 26 patients. High-spatial resolution MR imaging was performed in four cadaveric feet, and a prescribed imaging plane was used for depiction of the peroneal component of the PA. MR imaging delineated the anatomy of the PA and perifascial soft tissues. The peroneal component was best visualized in prescribed sagittal oblique images. Perifascial edema was the most common finding of plantar fasciitis, and it was remarkable in those cases with acute fascial rupture. MR imaging reliably delineated the anatomy of the PA and may allow precise localization and definition of the extent of involvement in disease processes. PMID- 11046172 TI - Invited commentary PMID- 11046171 TI - US of nerve entrapments in osteofibrous tunnels of the upper and lower limbs. AB - The diagnosis of nerve entrapment at osteofibrous tunnels relies primarily on clinical and electrodiagnostic findings. Recently, the refinement of high frequency broadband transducers with a range of 5-15 MHz, sophisticated focusing in the near field, and sensitive color and power Doppler technology have improved the ability to evaluate peripheral nerve entrapment in osteofibrous tunnels with ultrasonography (US). In the upper limb, osteofibrous tunnels amenable to US examination include the carpal tunnel for the median nerve and the cubital and Guyon tunnels for the ulnar nerve. In the lower limb, these tunnels include the fibular neck for the common peroneal nerve, the tarsal tunnel for the posterior tibial nerve, and the intermetatarsal spaces for the interdigital nerves. High resolution US allows direct imaging of the involved nerves, as well as documentation of changes in nerve shape and echotexture that occur in compressive syndromes. A spectrum of extrinsic causes of entrapment, such as tenosynovitis, ganglia, soft-tissue tumors, bone and joint abnormalities, and anomalous muscles, can also be diagnosed with US. With continued experience, it is likely that this technique will be increasingly used to evaluate nerve entrapment syndromes. PMID- 11046173 TI - Imaging of the painful lower limb stump. AB - Several postoperative complications associated with pain may develop in the stump of an amputated lower limb. Clinical findings are often nonspecific; however, radiologic evaluation, especially with magnetic resonance (MR) imaging, is useful in the early diagnosis of these complications, thereby helping minimize physical disability with its psychologic and socioeconomic implications. Conventional radiography can demonstrate evidence of osseous origins of pain (eg, aggressive bone edge, heterotopic ossification, osteomyelitis) and should be the first imaging study performed after clinical examination. Videofluoroscopy can help evaluate improper prosthetic fit by demonstrating abnormal residual limb motion, piston action, rolling of soft tissues, and abnormal angle between the limb axis and the prosthesis during gait. Ultrasonography can demonstrate inflammatory changes in the stump as well as soft-tissue fluid collections. However, MR imaging is the modality of choice when clinical and other imaging findings are indeterminate. Because of its high spatial and contrast resolution, MR imaging can demonstrate subtle inflammatory changes, fluid collections, cancers, neuromas, and subtle traumatic bone lesions. Knowledge of various surgical and rehabilitation techniques is required for accurate diagnosis of complications associated with stump pain. Correct diagnosis allows choice of the most appropriate therapy, which may involve treating the stump, remodeling the prosthesis, or both. PMID- 11046174 TI - Radiologic spectrum of craniocervical distraction injuries. AB - Injuries to the atlanto-occipital region, which range from complete atlanto occipital or atlantoaxial dislocation to nondisplaced occipital condyle avulsion fractures, are usually of critical clinical importance. At initial cross-table lateral radiography, measurement of the basion-dens and basion-posterior axial line intervals and comparison with normal measurements may help detect injury. Computed tomography (CT) with sagittal and coronal reformatted images permits optimal detection and evaluation of fracture and luxation. CT findings that may suggest atlanto-occipital injury include joint incongruity, focal hematomas, vertebral artery injury, capsular swelling, and, rarely, fractures through cranial nerve canals. Magnetic resonance (MR) imaging of the cervical spine with fat-suppressed gradient-echo T2-weighted or short-inversion-time inversion recovery sequences can demonstrate increased signal intensity in the atlantoaxial and atlanto-occipital joints, craniocervical ligaments, prevertebral soft tissues, and spinal cord. Axial gradient-echo MR images may be particularly useful in assessing the integrity of the transverse atlantal ligament. All imaging studies should be conducted with special attention to bone integrity and the possibility of soft-tissue injury. Atlanto-occipital injuries are now recognized as potentially survivable, although commonly with substantial morbidity. Swift diagnosis by the trauma radiologist is crucial for ensuring prompt, effective treatment and preventing delayed neurologic deficits in patients who survive such injuries. PMID- 11046175 TI - Radiography versus spiral CT in the evaluation of cervicothoracic junction injuries in polytrauma patients who have undergone intubation. AB - A prospective study was performed over a 1-year period in patients who had sustained blunt trauma, mostly in motor vehicle accidents. All 73 patients (56 male and 17 female; age range, 2-94 years; mean age, 35.2 years) in the study had undergone intubation and ventilation at the trauma site (mean Glasgow Coma Score, 9.9 [range, 3-15]; mean Injury Severity Score, 30.4 [range, 8-75]) and subsequently underwent three-view radiography of the cervical spine and thin section spiral computed tomography (CT) of the cervicothoracic junction. Spinal fractures were detected in 20 patients and involved the cervicothoracic junction region in 12 cases. In all 12 patients, the fractures were visualized at CT, whereas in seven of 12 patients, conventional radiography failed to demonstrate injuries (transverse process fracture of T1 [n = 1], pedicle and vertebral body fracture of C7 [n = 1], fractures of the first and second ribs [n = 5]). Thus, routine CT of the cervicothoracic junction in a highly select group of severely injured patients helped detect occult fracture in seven of 73 patients (10%); however, most of these fractures were not clinically significant. Larger studies involving a high-risk patient population are needed to confirm these findings. PMID- 11046176 TI - Invited commentary PMID- 11046177 TI - Invited commentary PMID- 11046178 TI - Nonseptic monoarthritis: imaging features with clinical and histopathologic correlation. AB - Diagnosis of septic arthritis requires aspiration and analysis of joint fluid. However, nonseptic articular disorders are fairly common and represent a significant diagnostic and therapeutic challenge. Such disorders include gout, Milwaukee shoulder, rapidly destructive articular disease, amyloid arthropathy, hemophilic arthropathy, primary synovial osteochondromatosis, pigmented villonodular synovitis, neuropathic arthropathy, and foreign-body synovitis. The clinical signs of articular disease, which include pain, swelling, and limitation of motion, are often nonspecific and can overlap with those of osseous or extraarticular disorders. Many articular processes have characteristic radiologic appearances that allow definitive diagnosis. Radiography is an important part of the evaluation of patients with articular disease. However, magnetic resonance (MR) imaging is the method of choice for characterizing the various disorders and assessing the full extent of osseous, chondral, and soft-tissue involvement. MR imaging can exquisitely demonstrate joint effusions, synovial proliferation, articular cartilage abnormalities, subchondral bone, ligaments, muscles, and juxtaarticular soft tissues. Although a wide spectrum of noninfectious processes may involve the joints, careful analysis of the imaging findings and correlation of these findings with the patient's clinical history can suggest a more specific diagnosis in most cases. Awareness and understanding of the underlying histopathologic findings aids in interpretation of MR images. PMID- 11046179 TI - Neuropathic osteoarthropathy: diagnostic dilemmas and differential diagnosis. AB - The purpose of this pictorial essay is to illustrate the radiologic spectrum of imaging findings of neuropathic osteoarthropathy. Typical findings include joint destruction, disorganization, and effusion with osseous debris. A variety of other imaging findings related to neuropathic osteoarthropathy such as resorption of the ends of tubular bones and neuropathic fracture are shown. The two prevailing theories for the pathophysiology of neuropathic bone and joint disease, the neurovascular and neurotraumatic theories, are briefly described. Examples of osteoarthropathy from diverse causes are presented including syringomyelia, spinal cord injury, meningomyelocele, diabetes mellitus, congenital insensitivity to pain, steroid injections, syphilis, leprosy, and others. The discussion focuses on key imaging features with emphasis on disease patterns and differential diagnosis, which vary by skeletal location. PMID- 11046181 TI - New directions for the journal of interventional cardiac electrophysiology PMID- 11046180 TI - Abnormal signal intensity in skeletal muscle at MR imaging: patterns, pearls, and pitfalls. AB - Abnormal signal intensity within skeletal muscle is frequently encountered at magnetic resonance (MR) imaging. Potential causes are diverse, including traumatic, infectious, autoimmune, inflammatory, neoplastic, neurologic, and iatrogenic conditions. Alterations in muscle signal intensity seen in pathologic conditions usually fall into one of three recognizable patterns: muscle edema, fatty infiltration, and mass lesion. Muscle edema may be seen in polymyositis and dermatomyositis, mild injuries, infectious myositis, radiation therapy, subacute denervation, compartment syndrome, early myositis ossificans, rhabdomyolysis, and sickle cell crisis. Fatty infiltration may be seen in chronic denervation, in chronic disuse, as a late finding after a severe muscle injury or chronic tendon tear, and in corticosteroid use. The mass lesion pattern may be seen in neoplasms, intramuscular abscess, myonecrosis, traumatic injury, myositis ossificans, muscular sarcoidosis, and parasitic infection. Some of these conditions require prompt medical or surgical management, whereas others do not benefit from medical intervention. The ability to accurately diagnose these conditions is therefore necessary, and biopsy may be required to establish the correct diagnosis. Clues to the correct diagnosis and whether biopsy is necessary or appropriate are often present on the MR images, especially when they are correlated with clinical features and the findings from other imaging modalities. PMID- 11046182 TI - Why a section on autonomics? PMID- 11046183 TI - A targeted disruption in connexin40 leads to distinct atrioventricular conduction defects. AB - INTRODUCTION: Gap junctions consist of connexin (Cx) proteins that enable electrical coupling of adjacent cells and propagation of action potentials. Cx40 is solely expressed in the atrium and His-Purkinje system. The purpose of this study was to evaluate atrioventricular (AV) conduction in mice with a homozygous deletion of Connexin40 (Cx40(-/-)). METHODS: Surface ECGs, intracardiac electrophysiology (EP) studies, and ambulatory telemetry were performed in Cx40( /-) mutant mice and wild-type (WT) controls. Atrioventricular (AV) conduction parameters and arrhythmia inducibility were evaluated using programmed stimulation. Analysis of heart rate variability was based on results of ambulatory monitoring. RESULTS: Significant findings included prolonged measures of AV refractoriness and conduction in connexin40-deficient mice, including longer PR, AH, and HV intervals, increased AV refractory periods, and increased AV Wenckebach and 2:1 block cycle lengths. Connexin40-deficient mice also had an increased incidence of inducible ventricular tachycardia, decreased basal heart rates, and increased heart rate variability. CONCLUSION: A homozygous disruption of Cx40 results in prolonged AV conduction parameters due to abnormal electrical coupling in the specialized conduction system, which may also predispose to arrhythmia vulnerability. PMID- 11046184 TI - Early ECG abnormalities associated with transcatheter closure of atrial septal defects using the Amplatzer septal occluder. AB - Conduction abnormalities and arrhythmias may occur in patients following secundum atrial septal defect (ASD) closure using the Amplatzer septal occluder (ASO). Therefore, the aim of this study was to prospectively perform ambulatory ECG monitoring to assess the electrocardiographic effects of transcatheter closure (TCC) of ASD using the ASO device. From 5/97 to 3/99, 41 patients with secundum ASD, underwent TCC using the ASO device at a median age of 9.2[emsp4 ]y. (0.5 87[emsp4 ]y.) and median weight of 34[emsp4 ]kg (5. 6-88[emsp4 ]kg.). Ambulatory Holter monitoring was performed pre- and immediately post TCC. Holter analysis included heart rate (HR), ECG intervals, supraventricular ectopy (SVE), ventricular ectopy (VE), and AV block. No change in baseline rhythm was noted in 37 patients (90%). Changes in AV conduction occurred in 3 patients (7%), including intermittent second degree AV block type II, and complete AV dissociation post closure. SVE was noted in 26 patients (63%) post closure, ranging from 5-2207 supraventricular premature beats (SVPB), including 9 patients (23%) with non-sustained supraventricular tachycardia (SVT), 3 of whom had short runs of SVT prior to closure. A significant increase in post-closure number of SVPB per hour (p=0.047) was noted. No significant difference was noted in PR interval, ventricular premature beats per hour, or QRS duration. CONCLUSIONS: Based on ambulatory ECG analysis, TCC of ASD with the ASO device is associated with an acute increase in SVE and a small risk of AV conduction abnormalities, including complete heart block. Long term follow-up studies will be necessary to determine late arrhythmia prevalence and relative frequency compared with standard surgical ASD repair. PMID- 11046185 TI - Recurrent seizure diagnosed by the insertable loop recorder. AB - INTRODUCTION: The Insertable Loop Recorder (ILR) has emerged as an important new tool in the diagnostic armamentarium for patients with syncope. METHODS AND RESULTS: A case report illustrates how the ILR unexpectedly led to the diagnosis of seizure as the explanation for a man's recurrent, but infrequent episodes of sudden loss of consciousness. CONCLUSIONS: This case raises the possibility that the development of implantable recording devices which monitor physiologic parameters other than cardiac rhythm (eg. brain, nerve or muscle activity) may provide the long-term monitoring capability needed to improve the diagnostic yield for conditions, such as seizures, which occur infrequently. PMID- 11046186 TI - Effects of sodium channel blockade on ibutilide induced prolongation of repolarization in the isolated rabbit ventricle. AB - INTRODUCTION: Ibutilide fumarate is indicated for the termination of atrial fibrillation and atrial flutter. It's mechanism of action is unclear but may involve activation of a late inward Na(+) current. METHODS AND RESULTS: Twenty seven experiments were performed using an isolated perfused rabbit right ventricle preparation. In each experiment effective refractory periods (ERP) and transmembrane 90% action potential durations (APD) were measured. In 8 experiments ERP and APD were measured at baseline, in the presence of ibutilide (0. 1[emsp4 ]uM), and in the presence of both ibutilide and tetrodotoxin (TTX, 2[emsp4 ]uM). In 8 experiments lidocaine (10[emsp4 ]uM) was used in place of TTX. Measures were made at 200, 400, and 800[emsp4 ]msec paced cycle lengths under each condition. The baseline values for APD at 200, 400 and 800[emsp4 ]msec cycle lengths for the experiments treated with ibutilide and TTX were 111+/-8, 140+/-14 and 159+/-22[emsp4 ]msec, respectively. In the presence of ibutilide, APD increased to 130+/-19, 192+/-26 and 217+/-35[emsp4 ]msec at 200, 400 and 800[emsp4 ]msec cycle lengths, respectively (all p< or =0.03). After the addition of TTX there was no shortening of APD or ERP compared to treatment with ibutilide alone at any cycle length (all p> or =0.062). Similarly, in the presence of ibutilide and lidocaine there were no changes in APD or ERP compared to treatment with ibutilide alone (all p > or =0.41). In 11 control experiments, there were no changes in APD or ERP on serial measures after placebo and TTX or lidocaine. CONCLUSION: Ibutilide induced prolongation of ventricular repolarization is not affected by Na(+) channel blockade with lidocaine or TTX in the isolated rabbit heart. These findings suggest that the effects of ibutilide are not mediated by a Na(+) channel dependent late current or that this mechanism contributes minimally to its action in this model. PMID- 11046187 TI - Demonstration of a His-Purkinje fatigue phenomenon with programmed stimulation of the right ventricular outflow tract. AB - This report describes the development of atrioventricular block by programmed stimulation of the right ventricular (RV) outflow tract in a patient with undiagnosed syncope. Burst pacing from the RV apex and outflow tract and programmed stimulation from the RV apex were unsuccessful. The observations were consistent with the fatigue phenomenon of the His-Purkinje system and illustrate the importance of an appropriate stimulation protocol in the electrophysiological evaluation of syncope. PMID- 11046188 TI - Is there an adverse outcome from abandoned pacing leads? AB - BACKGROUND: Indications for extraction of an abandoned pacemaker lead (APL) are controversial. The purpose of this study was to determine whether or not APLs should be extracted in the absence of pacemaker-related problems. METHODS AND RESULTS: We retrospectively reviewed, from 1977 through 1998, all patients with retained, non-functional leads and identified 433-266 males and 167 females. Mean age at initial pacemaker implantation was 68[emsp4 ]years. These patients received a total of 259 atrial and 948 ventricular leads. Of the total of 1,207 leads, 611 became non-functional. A total of 531 non-functional leads were abandoned, of which 18 were later extracted: one APL in 345 patients, two in 78, and three in 10. Indications for new lead placement when non-functional leads were abandoned included capture and/or sensing failure (243), lead recall (177), lead fracture (86), pacing system replacement to the contralateral side (11), accommodating patient growth (5), pacemaker function upgrade (5), replacement with implantable cardioverter defibrillator (ICD, 2), interference with ICD (1), and unknown (1). Complications that were associated with pacemakers were found in 24 patients (5.5%)-pacemaker system infection (8 patients) and venous occlusion at the time of a subsequent procedure of new lead placement when APLs had already been in place (16) which resulted in APL extraction (7) or transfer of the pacemaker system to the contralateral side (9). Neither venous thrombosis nor other complications were found in the remaining 409 patients (94.5%). The incidence of complications was higher in patients with three APLs than in patients with two or fewer APLs (40% vs. 4.7%, P=1x10(-6)), in patients with four or more total lead implantations than in patients with three or fewer total lead implantations (26.2% vs. 0. 6%, P<1x10(-10)), and in patients with three or more procedures of new lead placements than in patients with two or fewer procedures of new lead placements (36.4% vs. 3.9%, P=1x10(-10)). Patients with complications were younger than those without complications both at the time of initial pacemaker implantation (59+/-16 vs. 68+/-17 y, P=0.01) and when non-functional leads were abandoned (63+/-15 vs. 71+/-16 y, P=0.04). Mean numbers of APLs, total leads implanted, and procedures of new lead placement were significantly larger in patients with complications than in those without complications (1.58+/-0.78 vs. 1.2+/-0.44, 4.96+/-1.23 vs. 2.66+/-0.8, and 2.13+/-0.85 vs. 1.25+/-0.53, P=0.03, 4x10(-9) and 4x10(-5), respectively). CONCLUSIONS: 1. With only 5.5% of patients having had pacemaker-related complications, the adverse outcome of APL is small. 2. Clinical clues to the possible occasion for pacemaker-related complications include three or more APLs, four or more total leads, three or more procedures of new lead placement, and a younger age at initial pacemaker implantation. 3. Patients with a large number of APLs, total lead implantations, and procedures of new lead placement should be carefully observed to detect possible pacemaker-associated complications. PMID- 11046189 TI - Activation effects of single-site and dual-site right atrial pacing in canine. AB - Medical therapy for managing atrial fibrillation remains less than satisfactory. Electrical therapy such as right atrial (RA) pacing was shown to reduce rate of recurrence of atrial fibrillation, while evidently dual-site pacing was more effective than single-site pacing. However, similarities and/or differences in the electrophysiological consequences of single-site and dual-site RA pacing are unclear. Our objective was to simultaneously map RA and left atrial (LA) activation patterns and compare intra-atrial and interatrial activation properties during single-site and dual-site RA pacing in the normal canine heart. Basket-shaped catheters carrying 64 electrodes were deployed under the guidance of fluoroscopy and echocardiography into both the RA and LA of 7 dogs. Basket unipolar electrograms were simultaneously recorded while pacing at high lateral RA (HRA) alone, at inferior RA septum (RAS) alone, and at both sites simultaneously. We found that pacing at HRA alone resulted in the longest interatrial conduction time (47+/-6 ms). Pacing at RAS alone significantly shortened interatrial conduction time (29+/-5 ms) and completely activated both the RA and LA simultaneously (70+/-6 ms and 69+/-8 ms, respectively). Dual-site pacing at HRA and RAS significantly abbreviated RA complete activation time (52+/ 7 ms), but did not alter interatrial conduction time or LA activation pattern compared to pacing at RAS alone. In conclusion, single-site pacing at RAS shortened interatrial conduction time compared to HRA and completely activated both atria simultaneously in canines with normal atria. In addition to shortening interatrial conduction time, dual-site pacing at HRA and RAS abbreviated RA complete activation time. PMID- 11046190 TI - Threshold tracking pacing based on beat by beat evoked response detection: clinical benefits and potential problems. AB - Continuous monitoring of pacemaker stimulation thresholds and automatic adjustment of pacemaker outputs were among the longstanding goals of the pacing community. The first clinically successful implementation of threshold tracking pacing was the Autocapture feature which has accomplished automatic ventricular capture verification for every single stimulus by monitoring the Evoked Response (ER) signal resulting from myocardial depolarization. The Autocapture feature not only decreases energy consumption by keeping the stimulation output slightly above the actual threshold, but also increases patient safety by access to high output back-up pulses if there is loss of capture. Furthermore, it provides valuable documentation of stimulation thresholds over time and serves as a valuable research tool. Current limitations for its widespread use include the requirements for implantation of bipolar low polarization leads and unipolar pacing in the ventricle. Fusion/pseudofusion beats with resultant insufficient or even non-existent ER signal amplitudes followed by unnecessary delivery of back up pulses and a possible increase in pacemaker output is not an uncommon observation unique to the Autocapture feature. The recent incorporation of the Autocapture algorithm in dual chamber pacemakers has been challenging because of more frequent occurrence of fusion/pseudofusion beats in the presence of normal AV conduction. Along with a review of the previously published studies and our clinical experience, this article discusses the clinical advantages and potential problems of Autocapture. PMID- 11046191 TI - A novel indication for transvenous lead extraction: upgrading implantable cardioverter defibrillator systems. AB - Technological advances have resulted in the development of dual chamber pacemaker/defibrillator systems with smaller pectoral 'active cans'. Patients now have the option of upgrading from abdominal to pectoral or from single to dual chamber devices. In addition, due to the potential complications which may arise with abandoned ICD leads, extraction of preexisting leads may be preferable. METHODS AND RESULTS: Twenty consecutive patients (11 males), underwent lead extraction and upgrade, either from an abdominal to a pectoral, or from a single to a dual chamber device. The mean age was 62+/-18 years and mean implant duration was 50+/-14 months. Indications for extraction included lead fracture/malfunction (13), ERI/EOL (2), new SVT/VT (2), long charge times (2), and impending erosion (1). An initial attempt was made to remove the lead with gentle traction. If excessive scar tissue prohibited extraction, then a laser sheath was employed. Reimplantation proceeded following standard protocol. Clinical success was achieved in all patients. Eleven of thirty leads were removed with traction. The remaining 19 leads required removal with the laser sheath. All ICD reimplants were placed in the left pectoral position, of which 10 were dual chamber. The mean defibrillation threshold was 9.5+/-5.8 Joules. There were no procedure related perforations or deaths. At follow up (13+/-10 mos. ) there were no infections, lead malfunctions or venous thromboses. There were two deaths, both from intractable heart failure. CONCLUSIONS: This study demonstrates that, when indicated, ICD leads can be safely extracted and systems successfully upgraded to take advantage of new technology. PMID- 11046192 TI - Acute electrophysiologic effects and antifibrillatory actions of the long linear lesions in the right atrium in a sheep model. AB - Linear lesions (LL) represent an option for curing of atrial fibrillation (AF) with ablation techniques. METHODS AND RESULTS: In 11 sheep (w. 72+/-16 kg), LL were created with radiofrequency ablation in the lateral, posterior and septal walls of the right atrium (RA). AF was induced before and after LL with burst pacing. Mapping of the AF was performed with a 64-electrode basket catheter deployed in the RA. Quantitative analysis was performed with a custom-made software program. LL were confirmed histologically 7 to 10 days after the procedure. LL were transmural in 78% of their length. Stimulation thresholds and right atrial activation times were increased after LL compared to preablation values. Effective refractory periods of the RA were prolonged significantly in 7 out of 12 regions after generation of LL. Conduction velocities in the RA segments between LL were reduced in lateral, posterior and septal walls. During paced rhythms double potentials were recorded in all animals. AF could be induced in all animals of this model despite the presence of LL in the RA. AF episodes were significantly more regular after LL throughout the RA due to a significant reduction of the number of the wave fronts in the RA. During AF episodes, in the presence of LL, the RA was driven by wave fronts of left atrial origin entering the right side of the septum through interatrial connections. CONCLUSIONS: 1) LL profoundly affect electrophysiologic parameters of RA. 2) In the presence of LL, AF manifest a higher degree of regularity as compared to preablation episodes. 3) Dissociation between wave fronts of left atrial origin entering the RA through the interatrial connections is an important mechanism of the antifibrillatory action of the septal LL. PMID- 11046193 TI - Transcatheter cryoablation of ventricular myocardium in dogs. AB - INTRODUCTION: Surgical cryoablation, a highly effective technique used during antiarrhythmic surgery, produces voluminous, histologically uniform and discreet myocardial lesions. In contrast, radiofrequency (RF) catheter ablation, which as a result of its less invasive nature has largely supplanted antiarrhythmic surgery, produces smaller, histologically heterogeneous myocardial lesions. Since small lesion size and heterogeneity may reduce antiarrhythmic efficacy, we sought to reproduce the large, histologically homogeneous lesions created by surgical cryoablation, using a catheter cryoablation system (Cryogen, Inc., San Diego, CA) in the canine ventricle. METHODS AND RESULTS: In seven dogs, nineteen ventricular lesions (two right and seventeen left) were created with a 10F cryoablation catheter with either a 2 or 6 mm tip. In one dog AV node ablation was also performed. For each 'freeze', catheter tip nadir temperature, lesion width, depth, and transmurality were recorded, and lesion volume calculated. Average tip nadir temperature was -79.6+/-4.9 degrees C. Cooler nadir tip temperature was associated with deeper (p=.007) and more voluminous lesions (p=.042), and a greater likelihood of lesion transmurality (p=.034). Average lesion volume was 500+/-356 mm(3). No other variables predicted lesion volume or transmurality. Histologically, the catheter cryoablation lesions were sharply demarcated and homogeneous. The single freeze performed at the AV junction produced complete AV block. One complication, catheter rupture following its repetitive use, resulted in a coronary air embolus and death. CONCLUSION: Catheter cryoablation of canine ventricular myocardium produced voluminous, discrete, transmural lesions, which might be effective for ablation of ventricular tachycardia. Lesion volume and transmurality were dependent on catheter tip nadir temperature. PMID- 11046194 TI - As professionals, as people--we must be activists. PMID- 11046195 TI - The effects of intense physical exercise on secondary antibody response in young and old mice. AB - BACKGROUND AND PURPOSE: Based largely on data from young subjects, intense physical exercise is believed to suppress immune function. In addition, immune function, including secondary antibody response, declines with advancing age. Therefore, intense exercise in old subjects may further suppress the secondary antibody response. The purpose of this in vivo study was to investigate the effects of intense physical exercise on secondary antibody response in young (6-8 weeks) and old (22-24 months) C57BL/6 mice. SUBJECTS AND METHODS: Data were obtained from 22 young and 18 old C57BL/6 mice that were immunized to human serum albumin (HSA) and randomly divided into 3 groups. Two groups were exposed to a single bout of intense exercise to exhaustion and immediately boosted with an injection of HSA. The first group did not exercise further, but the second group continued with daily bouts of intense exercise to exhaustion for 9 days. The third group (control group) did not undergo intense exercise, but received the booster injection of HSA at the same time as the other groups. Ten days after the HSA booster injection, when high level of antibodies are produced in secondary antibody response, serum anti-HSA antibodies were measured by enzyme-linked immunosorbent assay. RESULTS: Young mice did not show suppression of secondary antibody response following intense exercise. However, old mice, exposed to a single bout of intense exercise, had an enhanced response similar to the response seen in young control mice. CONCLUSION AND DISCUSSION: The widely accepted hypothesis of immunosuppression resulting from intense exercise may not be true for old mice. PMID- 11046196 TI - Development of an activity scale for individuals with advanced Parkinson disease: reliability and "on-off" variability. AB - BACKGROUND AND PURPOSE: Functional mobility in people with advanced Parkinson disease, some of whom have a variable response to drug treatment, is often difficult to evaluate. The objectives of this study were to investigate the interrater reliability of measurements obtained with a scale designed to measure mobility and to determine the impact of self-rated dyskinesias and fluctuations on the measure. SSUBJECTS: Twenty-nine people with Parkinson disease and with disability and considerable disease duration (mean=11.7 years, SD=4.9, range=6 22) took part in the study. METHODS: The subjects' performance on a 10-item scale was videotaped. The videotapes were then scored by 2 independent raters, and the scores were used to determine interrater reliability. The stability of 6 repeated measurements was examined in the home situation, taking into account self-rated fluctuations of motor performance. RESULTS: Weighted Kappa values of agreement (.86-.98) confirmed the reliability between testers. Measurement during the "on" phase (when medication was working optimally) and the "off" phase (when the action of medication was strongly decreased or absent) led to different measurements. Measuring frequently within "on" and "off" phases gave relatively stable measurements for total function, bed transfers, and gait akinesia, the latter during the "off" phase only (intraclass correlation coefficients [ICCs]=.70-.93). However, more modest repeatability applied to transfers from a chair (ICC=.65-.67). CONCLUSION AND DISCUSSION: To ensure valid results in future effect studies, clinical differentiation between "on" and "off" phase measurements is proposed on the basis of patients' own perception of their medication status. PMID- 11046197 TI - Use of a classification system to guide nonsurgical management of a patient with chronic low back pain. AB - BACKGROUND AND PURPOSE: This case report describes the use of a classification system in the evaluation of a patient with chronic low back pain (LBP) and illustrates how this system was used to develop a management program in which the patient was instructed in symptom-reducing strategies for positioning and functional movement. CASE DESCRIPTION: The patient was a 55-year-old woman with a medical diagnosis of lumbar degenerative disk and degenerative joint disease from L2 to S1. Rotation with extension of the lumbar spine was found to be consistently associated with an increase in symptoms during the examination. Instruction was provided to restrict lumbar rotation and extension during performance of daily activities. OUTCOMES: The patient completed 8 physical therapy sessions over a 3-month period. Pretreatment, posttreatment, and 3-month follow-up modified Oswestry Disability Questionnaire scores were 43%, 16%, and 12%, respectively. DISCUSSION: Daily repetition of similar movements and postures may result in preferential movement of the lumbar spine in a specific direction, which then may contribute to the development, persistence, or recurrence of LBP. Research is needed to determine whether patients with LBP would benefit from training in activity modifications that are specific to the symptom-provoking movements and postures of each individual as identified through examination. PMID- 11046198 TI - Thirty-first Mary McMillan lecture. A time to harvest, a time to sow: ethics for a shifting landscape. PMID- 11046199 TI - Tipping the scales of time. PMID- 11046200 TI - Acidosis associated with perioperative saline administration: dilution or delusion? PMID- 11046201 TI - Role of pump prime in the etiology and pathogenesis of cardiopulmonary bypass associated acidosis. AB - BACKGROUND: The development of metabolic acidosis during cardiopulmonary bypass (CPB) is well recognized but poorly understood. The authors hypothesized that the delivery of pump prime fluids is primarily responsible for its development. Accordingly, acid-base changes induced by the establishment of CPB were studied using two types of priming fluid (Haemaccel, a polygeline solution, and Ringer's Injection vs. Plasmalyte 148) using quantitative biophysical methods. METHODS: A prospective, double-blind, randomized trial was conducted at a tertiary institution with 22 patients undergoing CPB for coronary artery bypass surgery. Sampling of arterial blood was performed at three time intervals: before CPB (t1), 2 min after initiation of CPB at full flows (t2), and at the end of the case (t3). Measurements of Na+, K+, Mg2+, Cl-, HCO3-, phosphate, Ca2+, albumin, lactate, and arterial blood gases at each collection point were performed. Results were analyzed in a quantitative manner. RESULTS: Immediately on delivery of pump prime fluids, all patients developed a metabolic acidosis (base excess: 0. 95 mEq/l (t1) to -3.65 mEq/l (t2) (P < 0.001) for Haemaccel-Ringer's and 1.17 mEq/l (t1) to -3.20 mEq/l (t2). The decrease in base excess was the same for both primes (-4.60 vs. -4.37; not significant). However, the mechanism of metabolic acidosis was different. With the Haemaccel-Ringer's prime, the metabolic acidosis was hyperchloremic (Delta Cl-, +9.50 mEq/l; confidence interval, 7.00-11.50). With Plasmalyte 148, the acidosis was induced by an increase in unmeasured anions, most probably acetate and gluconate. The resolution of these two processes was different because the excretion of chloride was slower than that of the unmeasured anions (Delta base excess from t1 to t3 = -1.60 for Haemaccel Ringer's vs. +1.15 for Plasmalyte 148; P = 0.0062). CONCLUSIONS: Cardiopulmonary bypass-induced metabolic acidosis appears to be iatrogenic in nature and derived from the effect of pump prime fluid on acid-base balance. The extent of such acidosis and its duration varies according to the type of pump prime. PMID- 11046202 TI - Acid-base changes caused by 5% albumin versus 6% hydroxyethyl starch solution in patients undergoing acute normovolemic hemodilution: a randomized prospective study. AB - BACKGROUND: Preoperative acute normovolemic hemodilution (ANH) is an excellent model for evaluating the effects of different colloid solutions that are free of bicarbonate but have large chloride concentrations on acid-base equilibrium. METHODS: In 20 patients undergoing gynecologic surgery, ANH to a hematocrit of 22% was performed. Two groups of 10 patients each were randomly assigned to receive either 5% albumin or 6% hydroxyethyl starch solutions containing chloride concentrations of 150 and 154 mm, respectively, during ANH. Blood volume (double label measurement of plasma and red cell volumes), pH, Paco2, and serum concentrations of sodium, potassium, chloride, lactate, ionized calcium, phosphate, albumin, and total protein were measured before and 20 min after completion of ANH. Strong ion difference was calculated as serum sodium plus serum potassium minus serum chloride minus serum lactate. The amount of weak plasma acid was calculated using a computer program. RESULTS: After ANH, blood volume was well maintained in both groups. ANH caused slight metabolic acidosis with hyperchloremia and a concomitant decrease in strong ion difference. Plasma albumin concentration decreased after hemodilution with 6% hydroxyethyl starch solution and increased after hemodilution with 5% albumin solution. Despite a three-times larger decrease in strong ion difference after ANH with 6% hydroxyethyl starch solution, the decrease in pH was nearly the same in both groups. CONCLUSIONS: ANH with 5% albumin or 6% hydroxyethyl starch solutions led to metabolic acidosis. A dilution of extracellular bicarbonate or changes in strong ion difference and albumin concentration offer explanations for this type of acidosis. PMID- 11046203 TI - Dilutional acidosis following hetastarch or albumin in healthy volunteers. AB - BACKGROUND: The intent of this study was to evaluate the impact of the commonly used colloids-hetastarch and albumin-on in vivo acid-base balance. From this evaluation, a better understanding of the mechanism of dilutional acidosis was expected. METHODS: In a prospective, randomized fashion, 11 healthy volunteers were administered 15 ml/kg hetastarch solution, 6%, or 15 ml/kg albumin, 5%, intravenously over 30 min. Four weeks later, the study subjects were administered the other colloid. Arterial blood gas and electrolyte parameters were measured at baseline and at 30, 60, 90, 120, 210, and 300 min after colloid administration. Pre- and postlaboratory values were compared within groups using a paired t test and a Wilcoxon signed rank test and between groups using repeated-measures analysis of variance and a Wilcoxon rank sum test. RESULTS: Thirty min after infusion, subjects who were administered hetastarch showed statistically significant changes (P < 0.05) in base excess (from 2.5 +/- 0.9 mEq/l to 0.7 +/- 1.1 mEq/l), HCO3- concentration (from 27 +/- 1.0 mEq/l to 25 +/- 1.3 mEq/l), Cl- concentration (from 108 +/- 2 mEq/l to 112 +/- 2 mEq/l), albumin concentration (from 4.4 +/- 0.2 g/dl to 3.5 +/- 0.5 g/dl), and arterial carbon dioxide tension (Paco2; from 40.8 +/- 2.3 mmHg to 39. 2 +/- 3.2 mmHg), whereas only the albumin concentration (from 4.4 +/- 0.2 g/dl to 4.8 +/- 0.6 g/dl) changed significantly in the albumin-treated group. CONCLUSIONS: Decreases in base excess were observed for 210 min after hetastarch administration but not after albumin. The mechanism for this difference is discussed. PMID- 11046204 TI - Minimum alveolar concentration-awake of Xenon alone and in combination with isoflurane or sevoflurane. AB - BACKGROUND: The minimum alveolar concentration (MAC)-awake is a traditional index of hypnotic potency of an inhalational anesthetic. The MAC-awake of xenon, an inert gas with anesthetic properties (MAC = 71%), has not been determined. It is also unknown how xenon interacts with isoflurane or sevoflurane on the MAC-awake. METHODS: In the first part of the study, 90 female patients received xenon, nitrous oxide (N2O), isoflurane, or sevoflurane supplemented with epidural anesthesia (n = 36 for xenon and n = 18 per group for other anesthetics). In the second part, 72 additional patients received either xenon or N2O combined with the 0.5 times MAC-awake concentration of isoflurane or sevoflurane (0.2% and 0.3%, respectively, based on the results of the first part; n = 18 per group). During emergence, the concentration of an assigned anesthetic (xenon or N2O only in the second part) was decreased in 0. 1 MAC decrements every 15 min from 0.8 MAC or from 70% in the case of N2O until the patient followed the command to either open her eyes or to squeeze and release the investigator's hand. The concentration midway between the value permitting the first response to command and that just preventing it was defined as the MAC-awake. RESULTS: The MAC-awake were as follows: xenon, 32.6 +/- 6.1% (mean +/- SD) or 0.46 +/- 0.09 MAC; N2O, 63.3 +/- 7.1% (0.61 +/- 0.07 MAC); isoflurane, 0.40 +/- 0.07% (0.35 +/- 0.06 MAC); and sevoflurane, 0.59 +/- 0.10% (0.35 +/- 0.06 MAC). Addition of the 0.5 MAC-awake concentrations of isoflurane and sevoflurane reduced the MAC-awake of xenon to 0.50 +/- 0.15 and 0.51 +/- 0.16 times its MAC-awake as a sole agent, but that of N2O to the values significantly greater than 0.5 times its MAC-awake as a sole agent (0.68 +/- 0.12 and 0.66 +/- 0.14 times MAC-awake; P < 0.01, analysis of variance and Dunnett's test). CONCLUSIONS: The MAC-awake of xenon is 33% or 0.46 times its MAC. In terms of the MAC-fraction, this is smaller than that for N2O but greater than those for isoflurane and sevoflurane. Unlike N2O, xenon interacts additively with isoflurane and sevoflurane on MAC-awake. PMID- 11046205 TI - Neuromuscular monitoring at the orbicularis oculi may overestimate the blockade in myasthenic patients. AB - BACKGROUND: In most publications about myasthenia, monitoring neuromuscular blockade during anesthesia is recommended. In healthy patients, the relation of blockade between muscles has been established, but there is little information about the relation in myasthenic patients. Our objective was to investigate whether the relation between the orbicularis oculi and adductor pollicis muscles is the same in healthy patients and myasthenic patients. METHODS: After anesthesia was induced with 4-6 mg/kg thiopental and 2 microg/kg fentanyl, followed by 2% sevoflurane and 60% nitrous oxide in oxygen, 10 healthy patients and 10 myasthenic patients received 0. 025 and 0.01 mg/kg vecuronium, respectively. Neuromuscular monitoring was performed with use of accelerometry at the orbicularis oculi and the adductor pollicis muscles by stimulating the temporal branch of the facial nerve and the ulnar nerve. RESULTS: The relation of blockade between these two muscles was not the same in healthy patients and myasthenic patients: in healthy patients, the maximum neuromuscular blockade with 0.025 mg/kg vecuronium was less in the orbicularis oculi than in the adductor pollicis (median 72% vs. 91%; P < 0.05); in contrast, in myasthenic patients, the blockade with 0.01 mg/kg vecuronium was greater in the orbicularis oculi than in the adductor pollicis (median 96% vs. 62%; P < 0.05). CONCLUSION: Neuromuscular monitoring at the orbicularis oculi may overestimate blockade in myasthenic patients. Extubation must be performed when the muscle most sensitive to neuromuscular blocking agents is recovered. Therefore, neuromuscular monitoring at the orbicularis oculi is recommended to avoid persistent neuromuscular blockade in patients with myasthenia gravis. PMID- 11046206 TI - Inhaled albuterol, but not intravenous lidocaine, protects against intubation induced bronchoconstriction in asthma. AB - BACKGROUND: The ability of intravenous lidocaine to prevent intubation-induced bronchospasm is unclear. The authors performed a prospective, randomized, double blind, placebo-controlled trial to test the ability of intravenous lidocaine and inhaled albuterol to attenuate airway reactivity after tracheal intubation in asthmatic patients undergoing general anesthesia. METHODS: Sixty patients were randomized to receive either 1.5 mg/kg intravenous lidocaine or saline, 3 min before tracheal intubation. An additional 50 patients were randomized to receive 4 puffs of inhaled albuterol or placebo 15-20 min before tracheal intubation. Anesthesia was induced with propofol. Immediately after intubation and at 5-min intervals, transpulmonary pressure and airflow were recorded, and lower pulmonary resistance (RL) was calculated. Isoflurane was administered after the initial two measurements to assess reversibility of bronchoconstriction. A bronchoconstrictor response to intubation was defined as RL greater than or equal to 5 cm H2O. l-1. s-1 in the first two measurements after intubation and RL subsequently decreasing by 50% or more after isoflurane. RESULTS: The lidocaine and placebo groups were not different in the peak RL before administration of isoflurane (8.2 cm H2O. l 1. s-1 vs. 7.6 cm H2O. l-1. s-1) or frequency of airway response to intubation (lidocaine 6 of 30 vs. placebo 5 of 27). In contrast, the albuterol group had lower peak RL (5.3 cm H2O. l-1. s-1 vs. 8.9 cm H2O. l-1. s-1; P < 0.05) and a lower frequency of airway response (1 of 25 vs. 8 of 23; P < 0.05) than the placebo group. CONCLUSIONS: Inhaled albuterol blunted airway response to tracheal intubation in asthmatic patients, whereas intravenous lidocaine did not. PMID- 11046207 TI - Graded hypercapnia and cerebral autoregulation during sevoflurane or propofol anesthesia. AB - BACKGROUND: Hypercapnia abolishes cerebral autoregulation, but little is known about the interaction between hypercapnia and autoregulation during general anesthesia. With normocapnia, sevoflurane (up to 1.5 minimum alveolar concentration) and propofol do not impair cerebral autoregulation. This study aimed to document the level of hypercapnia required to impair cerebral autoregulation during propofol or sevoflurane anesthesia. METHODS: Eight healthy subjects received a remifentanil infusion and were anesthetized with propofol (140 microg. kg-1. min-1) and sevoflurane (1.0-1.1% end tidal) in a randomized crossover study. Ventilation was adjusted to achieve incremental increases in arterial carbon dioxide partial pressure (Paco2) until autoregulation was impaired. Cerebral autoregulation was tested by increasing the mean arterial pressure (MAP) from 80 to 100 mmHg with phenylephrine while measuring middle cerebral artery flow velocity by transcranial Doppler. The autoregulation index, which has a value ranging from 0 to 1, representing absent to perfect autoregulation, was calculated, and an autoregulation index of 0.4 or less represented significantly impaired autoregulation. RESULTS: The threshold Paco2 to significantly impair cerebral autoregulation ranged from 50 to 66 mmHg. The threshold averaged 56 +/- 4 mmHg (mean +/- SD) during sevoflurane anesthesia and 61 +/- 4 mmHg during propofol anesthesia (P = 0.03). Carbon dioxide reactivity measured at a MAP of 100 mmHg was 30% greater than that at a MAP of 80 mmHg. CONCLUSIONS: Even mild hypercapnia can significantly impair cerebral autoregulation during general anesthesia. There is a significant difference between propofol anesthesia and sevoflurane anesthesia with respect to the effect of hypercapnia on cerebral autoregulation. This difference occurs at clinically relevant levels of Paco2. When inducing hypercapnia, carbon dioxide reactivity is significantly affected by the MAP. PMID- 11046208 TI - Plasma and urinary cytokine homeostasis and renal dysfunction during cardiac surgery. AB - BACKGROUND: Cardiac surgery induces changes in plasma cytokines. Proinflammatory cytokines have been associated with a number of renal diseases. The proinflammatory cytokines interleukin 8 (IL-8), tumor necrosis factor alpha (TNFalpha), and interleukin 1beta (IL-1beta) are smaller than the antiinflammatory cytokines interleukin 10 (IL-10), interleukin 1 receptor antagonist (IL-1ra), and TNF soluble receptor 2 (TNFsr2), and thus undergo glomerular filtration more readily. Accordingly, this study investigated the relation between plasma and urinary cytokines and proximal renal dysfunction during cardiac surgery. METHODS: Twenty patients undergoing coronary artery bypass grafting with cardiopulmonary bypass (CPB) were studied. Blood and urine samples were analyzed for proinflammatory and antiinflammatory cytokines. Proximal tubular dysfunction was measured using urinary N-acetyl-beta-d glucosaminidase (NAG)/creatinine and alpha1-microglobulin/creatinine ratios. RESULTS: Plasma IL-8, IL-10, IL-1ra, and TNFsr2 values were significantly elevated compared with baseline. Urinary IL-1ra and TNFsr2 were significantly elevated. Urinary NAG/creatinine and alpha1-microglobulin/creatinine ratios were also elevated. Plasma TNFalpha at 2 h correlated with urinary NAG/creatinine ratio at 2 and 6 h (P < 0.05) and with urinary IL-1ra at 2 h (P < 0.05). Plasma IL-8 at 2 h correlated with NAG/creatinine at 6 h (P < 0.05). Urinary IL-1ra correlated with urinary NAG/creatinine ratio after cross-clamp release and 2 and 6 h after CPB (P < 0.05). CONCLUSIONS: Cardiac surgery using CPB leads to changes in plasma and urinary cytokine homeostasis that correlate with renal proximal tubular dysfunction. This dysfunction may be related to the renal filtration of proinflammatory mediators. Renal autoprotective mechanisms may involve the intrarenal generation of antiinflammatory cytokines. PMID- 11046209 TI - Reevaluation of rectal ketamine premedication in children: comparison with rectal midazolam. AB - BACKGROUND: Results of previous studies of rectal ketamine as a pediatric premedication are clouded because of lack of dose-response relation, inappropriate time of assessing sedative effects, and previous administration or coadministration of benzodiazepines. Therefore, the authors reevaluated the efficacy of rectally administered ketamine in comparison with 1 mg/kg rectal midazolam. METHODS: Sixty-six infants and children (age, 7-61 months) who were American Society of Anesthesiologists physical status I and who were undergoing minor surgeries as in-patients were randomized to receive 5 mg/kg ketamine (n = 16), 7 mg/kg ketamine (n = 16), 10 mg/kg ketamine (n = 17), or 1 mg/kg midazolam (n = 17) via rectum. A blinded observer scored sedation 45 min and 15 min after administration of ketamine and midazolam, respectively, when children were separated from parent(s) for inhalational induction. All children underwent standardized general anesthesia with sevoflurane, nitrous oxide, and oxygen with endotracheal intubation. Blood pressure, heart rate, and oxyhemoglobin saturation were determined before, during, and after anesthesia. Postoperative recovery characteristics and incidence of adverse reactions were also assessed. RESULTS: Most children (88%) who received rectally 10 mg/kg ketamine or 1 mg/kg midazolam separated easily from their parents compared with those (31%) who received 7 or 5 mg/kg rectal ketamine (P < 0.05). Similarly, more children who received 10 mg/kg ketamine or 1 mg/kg midazolam underwent mask induction without struggling or crying compared with those who received 7 or 5 mg/kg ketamine (P < 0.05). There were no clinically significant changes in blood pressure, heart rate, and oxyhemoglobin saturation after administration of either drug. Immediately after surgery, more children receiving midazolam or 5 mg/kg ketamine were agitated compared with 7 or 10 mg/kg ketamine. Ketamine, 7 and 10 mg/kg, provided postoperative analgesia, but the largest dose of ketamine was associated with delayed emergence from general anesthesia. CONCLUSIONS: The results indicate that rectally administered ketamine alone produces dose-dependent sedative effects in children, when evaluated at its predicted peak plasma concentration. Ketamine, 10 mg/kg, has a delayed onset but is as effective as 1 mg/kg midazolam for sedating healthy children before general anesthesia. However, 10 mg/kg rectal ketamine is not recommended for brief surgeries because of prolonged postoperative sedation. PMID- 11046210 TI - Comparison of the costs and recovery profiles of three anesthetic techniques for ambulatory anorectal surgery. AB - BACKGROUND: Given the current practice environment, it is important to determine the anesthetic technique with the highest patient acceptance and lowest associated costs. The authors compared three commonly used anesthetic techniques for anorectal procedures in the ambulatory setting. METHODS: Ninety-three consenting adult outpatients undergoing anorectal surgery were randomly assigned to one of three anesthetic treatment groups: group 1 received local infiltration with a 30-ml mixture containing 15 ml lidocaine, 2%, and 15 ml bupivacaine, 0.5%, with epinephrine (1:200,000) in combination with intravenous sedation using a propofol infusion, 25-100 microg. kg-1. min-1; group 2 received a spinal subarachnoid block with a combination of 30 mg lidocaine and 20 microg fentanyl with midazolam, 1-2-mg intravenous bolus doses; and group 3 received general anesthesia with 2.5 mg/kg propofol administered intravenously and 0.5-2% sevoflurane in combination with 65% nitrous oxide. In groups 2 and 3, the surgeon also administered 10 ml of the previously described local anesthetic mixture at the surgical site before the skin incision. RESULTS: The mean costs were significantly decreased in group 1 ($69 +/- 20 compared with $104 +/- 18 and $145 +/- 25 in groups 2 and 3, respectively) because both intraoperative and recovery costs were lowest (P < 0.05). Although the surgical time did not differ among the three groups, the anesthesia time and times to oral intake and home-readiness were significantly shorter in group 1 (vs. groups 2 and 3). There was no significant difference among the three groups with respect to the postoperative side effects or unanticipated hospitalizations. However, the need for pain medication was less in groups 1 and 2 (19% and 19% vs. 45% for group 3; P < 0.05). Patients in group 1 had no complaints of nausea (vs. 3% and 26% in groups 2 and 3, respectively). More patients in group 1 (68%) were highly satisfied with the care they received than in groups 2 (58%) and 3 (39%). CONCLUSIONS: The use of local anesthesia with sedation is the most cost-effective technique for anorectal surgery in the ambulatory setting. PMID- 11046211 TI - Low- and medium-molecular-weight hydroxyethyl starches: comparison of their effect on blood coagulation. AB - BACKGROUND: High-molecular-weight hydroxyethyl starch (HES) compromises blood coagulation more than medium-molecular-weight HES. The authors compared medium molecular weight HES (200 kd [HES200]) and low-molecular-weight HES (70 kd [HES70]). METHODS: In a prospective, double-blind, randomized-sequence crossover study, 22 male volunteers received 15 ml/kg HES200 and HES70. Blood samples were taken before and 5 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 24 h after infusion. The following parameters were analyzed at all time points: prothrombin time, activated partial thromboplastin time, fibrinogen, factor VIII, antigenetic and functional von Willebrand factor, platelets, Thrombelastograph analysis parameters (reaction time, coagulation time, maximum amplitude, angle alpha, and clot lysis 30 and 60 min after maximum amplitude), ionized calcium, hematocrit, HES plasma concentration, molecular weight (weight average and number average), molar substitution, and polydispersity (weight average/number average). Repeated measures analysis of variance (P < 0.05) was used to compare the response of the aforementioned parameters to the infusion of HES70 and HES200. RESULTS: Both HES solutions had a significant impact on all parameters. A slightly greater compromise with HES200 was found in activated partial thromboplastin time (P = 0.010), factor VIII (P = 0.009), antigenetic von Willebrand factor (P = 0.041), functional von Willebrand factor (P = 0.026), maximum amplitude (P = 0.008), and angle alpha (P = 0.003). No difference was established with the other parameters. HES concentration (P < 0.001), weight average (P < 0.001), number average (P < 0.001), and polydispersity (P < 0.001) were higher with HES200. There was no difference with molar substitution (P = 0.091). CONCLUSIONS: Low-molecular-weight hydroxyethyl starch (70 kd) compromises blood coagulation slightly less than HES200, but it is unclear whether this is clinically relevant. PMID- 11046212 TI - Effect of patient-triggered ventilation on respiratory workload in infants after cardiac surgery. AB - BACKGROUND: Patient-triggered ventilation (PTV) is commonly used in adults to avoid dyssynchrony between patient and ventilator. However, few investigations have examined the effects of PTV in infants. Our objective was to determine if pressure-control PTV reduces infants' respiratory workloads in proportion to the level of pressure control. We also explored which level of pressure control provided respiratory workloads similar to those after the extubation of the trachea. METHODS: When seven post-cardiac surgery infants, aged 1 to 11 months, were to be weaned with the pressure-control PTV, we randomly applied five levels of pressure control: 0, 4, 8, 12, and 16 cm H2O. All patients were ventilated with assist-control mode, triggering sensitivity of 1 l/min, and positive end expiratory pressure of 3 cm H2O. After establishing steady state conditions at each level of pressure control, arterial blood gases were analyzed and esophageal pressure (Pes), airway pressure, and airflow were measured. Inspiratory work of breathing (WOB) was calculated using a Campbell diagram. A modified pressure-time product (PTPmod) and the negative deflection of Pes were calculated from the Pes tracing below the baseline. The measurement was repeated after extubation. RESULTS: Pressure-control PTV supported every spontaneous breath. By decreasing the level of pressure control, respiratory rate increased, tidal volume decreased, and as a result, minute ventilation and arterial carbon dioxide partial pressure were maintained stable. The WOB, PTPmod, and negative deflection of Pes increased as pressure control level was decreased. The WOB and PTPmod at 4 cm H2O pressure control and 0 cm H2O pressure control and after extubation were significantly greater than those at the pressure control of 16, 12, and 8 cm H2O (P < 0.05). The WOB and PTPmod were almost equivalent after extubation and at 4 cm H2O pressure control. CONCLUSIONS: Work of breathing and PTPmod were changed according to the pressure control level in post-cardiac surgery infants. PTV may be feasible in infants as well as in adults. PMID- 11046213 TI - Sex differences in morphine analgesia: an experimental study in healthy volunteers. AB - BACKGROUND: Animal and human studies indicate the existence of important sex related differences in opioid-mediated behavior. In this study the authors examined the influence of morphine on experimentally induced pain in healthy male and female volunteers. METHODS: Young healthy men and women (10 of each sex) received intravenous morphine (bolus 0.1-mg/kg dose followed by an infusion of 0.030 mg. kg-1. h-1 for 1 h). Pain threshold and pain tolerance in response to a gradual increase in transcutaneous electrical stimulation, as well as plasma concentrations of morphine and its major metabolites (morphine-6-glucuronide and morphine-3-glucuronide) were determined at regular intervals up to 7 h after the start of morphine infusion. A population pharmacodynamic model was used to analyze the morphine-induced changes in stimulus intensity. The improvement of the model fits by inclusion of covariates (sex, age, weight, lean body mass) was tested for significance. The model is characterized by baseline current, a rate constant for equilibrium between plasma and effect-site morphine concentrations (ke0), and analgesic potency (AC50, or the morphine concentration causing a 100% increase in stimulus intensity for response). RESULTS: The inclusion of the covariates age, weight, and lean body mass did not improve the model fits for any of the model parameters. For both pain threshold and tolerance, a significant dependency on sex was observed for the parameters ke0 (pain threshold: 0.0070 +/- 0.0013 (+/- SE) min-1 in men vs. 0.0030 +/- 0. 0005 min-1 in women; pain tolerance: 0.0073 +/- 0.0012 min-1 in men vs. 0.0024 +/- 0.0005 min-1 in women) and AC50 (pain threshold: 71.2 +/- 10.5 nm in men vs. 41.7 +/- 8.4 nm in women; pain tolerance: 76. 5 +/- 7.4 nm in men vs. 32.9 +/- 7.9 nm in women). Baseline currents were similar for both sexes: 21.4 +/- 1.6 mA for pain threshold and 39.1 +/- 2.3 mA for pain tolerance. Concentrations of morphine, morphine-3 glucuronide, and morphine-6-glucuronide did not differ between men and women. CONCLUSIONS: These data show sex differences in morphine analgesia, with greater morphine potency but slower speed of onset and offset in women. The data are in agreement with observations of sex differences in morphine-induced respiratory depression and may explain higher postoperative opioid consumption in men relative to women. PMID- 11046214 TI - Ventilator-associated sinusitis: microbiological results of sinus aspirates in patients on antibiotics. AB - BACKGROUND: The efficacy of systemic antibiotics on the treatment of ventilator associated infectious maxillary sinusitis (VAIMS) is debated. The objective of this study was to determine the etiologic diagnosis of VAIMS in patients receiving antibiotics. METHODS: Patients mechanically ventilated for more than or equal to 72 h, who had persistent fever while on antibiotics for more than or equal to 48 h, underwent computed tomography scan followed by transnasal puncture of involved maxillary sinuses. VAIMS was defined as follows: fever greater than or equal to 38 degrees C, radiographic signs (air fluid level or opacification of maxillary sinuses on computed tomography scan), and a quantitative culture of sinus aspirate yielding more than or equal to 103 colony-forming units/ml. RESULTS: Twenty-four patients had radiographic signs of sinusitis. The mean +/- SD prior durations of mechanical ventilation and antibiotic exposure were 9.5 +/- 4.7 days and 6 +/- 4 days, respectively. Six unilateral and nine bilateral VAIMS were diagnosed in 15 patients. The median number of etiologic organisms per patient was two (range, one to four). The bacteriologic cultures yielded gram positive bacteria (n = 21), gram-negative bacteria (n = 22), and yeasts (n = 5). Forty percent of causative agents were susceptible to the antibiotics prescribed. Seven patients with VAIMS developed 10 concomitant infections: ventilator associated pneumonia (n = 5), urinary tract infection (n = 3), catheter infections (n = 2). In all cases of ventilator-associated pneumonia, the implicated agents were the causative agents of VAIMS. CONCLUSION: In VAIMS patients on antibiotics, quantitative cultures of sinus aspirates may contribute to establish the diagnosis. The frequent recovery of microorganisms susceptible to the antimicrobial treatment administered suggests that therapy of VAIMS with systemic antibiotics may not be sufficient. PMID- 11046215 TI - Compatibility of different colloid plasma expanders with perflubron emulsion: an intravital microscopic study in the hamster. AB - BACKGROUND: Perfluorocarbon-based oxygen carriers have been proposed as an adjunct to autologous blood conservation techniques during elective surgery. To date, the effects of perfluorocarbon emulsions at the microcirculatory level have not been studied extensively. In this study the effects of perflubron emulsion on the microcirculation after acute normovolemic hemodilution (ANH) were investigated using different colloid plasma expanders. METHODS: The dorsal skin fold chamber model and intravital fluorescence microscopy were used for analysis of the microcirculation in the thin striated skin muscle of conscious hamsters (body weight, 40-60 g). Measurements of microvascular perfusion and leukocyte adhesion (n = 6 animals per experimental group) were made before and at 10, 30, and 60 min after ANH (to hematocrit 0.3) with either 6% hydroxyethyl starch 200/0.6 (HES), 3.5% gelatin, 5% human serum albumin (HSA), or 6% dextran 60 (DX 60) followed by intravenous injection of 3 ml/kg body weight of a 60% weight/volume perfluorocarbon emulsion based on perflubron (perfluorooctyl bromide) emulsified with egg yolk lecithin. RESULTS: Acute normovolemic hemodilution with HES, gelatin, or HSA followed by injection of perflubron emulsion elicited no alterations of local microvascular perfusion or leukocyte endothelium interaction as assessed in arterioles and postcapillary venules. However, ANH with DX-60 followed by injection of perflubron emulsion led to a significant reduction of erythrocyte velocity in postcapillary venules and an increase in venular leukocyte sticking that was never observed with DX-60 alone. CONCLUSIONS: Hydroxyethyl starch, gelatin, and HSA are compatible with perflubron emulsion in the setting of ANH. Only DX-60 appeared to be incompatible with perflubron emulsion, as evidenced by impairment of capillary perfusion. PMID- 11046216 TI - Protection of sensory function and antihyperalgesic properties of a prosaposin derived peptide in diabetic rats. AB - BACKGROUND: Short-term diabetes causes sensory disorders in rats ranging from thermal hypoalgesia to exaggerated behavioral responses to other sensory stimuli. As impaired neurotrophic support may promote sensory nerve disorders during diabetes, the authors investigated whether TX14(A), a neurotrophic peptide derived from prosaposin, was able to ameliorate nerve disorders in diabetic rats. METHODS: TX14(A) was delivered by intraperitoneal or intrathecal injection to control or streptozotocin-diabetic rats in either single or multiple (three times weekly) dose regimens. Efficacy was measured against diabetes-induced disorders of sensory nerve conduction velocity, paw withdrawal latency to radiant heat, tactile response thresholds to von Frey filaments, and flinching after paw formalin injection. RESULTS: Prolonged TX14(A) treatment of diabetic rats prevented the progressive decline in large sensory fiber conduction velocity in the sciatic nerve, development of paw thermal hypoalgesia, and increased flinching after paw formalin injection. The effect on formalin hyperalgesia persisted for 48 h but not 72 h after injection. No effects were noted in control rats. A single injection of TX14(A) 30 min before testing did not alter thermal response latencies in control or diabetic rats but prevented formalin hyperalgesia in diabetic rats. Tactile allodynia and the prolonged paw thermal hyperalgesia to radiant heat after intrathecal delivery of substance P were also dose-dependently ameliorated in diabetic rats by a single injection of TX14(A), whereas no effects were observed on the responses to these tests in control rats. CONCLUSIONS: TX14(A) exhibits both neuroprotective and acute antihyperalgesic properties in diabetic rats without altering normal nociceptive function. PMID- 11046217 TI - The effect of chronic dexamethasone-induced hyperglycemia and its acute treatment with insulin on brain glucose and glycogen concentrations in rats. AB - BACKGROUND: In the rat model of forebrain ischemia, long-term dexamethasone treatment is reported to cause hyperglycemia and worsen postischemic functional and histologic injury. This effect was assumed to result from glucose enhancement of intraischemic lactic acidosis within the brain. Short-term insulin therapy restored normoglycemia but did not return histologic injury completely to baseline values. Using a nonischemic rat model, the current study attempted to identify a metabolic basis for such outcome data. METHODS: Fifty-eight halothane anesthetized (1.3% inspired) Sprague-Dawley rats were assigned randomly to be administered either no treatment (N = 18) or 2 mg/kg intraperitoneal dexamethasone (N = 40). The latter were administered dexamethasone 3 h before the study only (N = 8) or for 3 h before the study plus daily for 1 day (N = 8), 2 days (N = 8), or 4 days (N = 16). Of the rats treated with dexamethasone for 4 days, one half (N = 8) were administered an insulin-containing saline infusion subsequently to restore normoglycemia short-term. All other rats (N = 50) were administered an infusion of saline without insulin. Plasma glucose was quantified, and brains were excised after in situ freezing. Brain glucose and glycogen concentrations were measured using enzymatic fluorometric analyses. RESULTS: After 4 days of dexamethasone treatment, plasma glucose was 159% greater than in rats administered placebo (i.e., 22.01 +/- 4.66 vs. 8.51 +/- 1.65 micromol/ml; mean +/- SD; P < 0.0001). Brain glucose concentrations increased parallel to plasma glucose. An insulin infusion for 27 +/- 5 min restored normoglycemia but resulted in a brain-to-plasma glucose ratio that was 32% greater than baseline values (P < 0.01). Neither dexamethasone nor the combination of dexamethasone plus insulin affected brain glycogen concentrations. CONCLUSIONS: In a nonischemic rat model, dexamethasone alone had no independent effect on the brain-to-plasma glucose ratio. However, short-term insulin therapy caused a dysequilibrium between plasma and brain glucose, resulting in an underestimation of brain glucose concentrations when normoglycemia was restored. The dysequilibrium likely was caused by the rapid rate of glucose reduction. The magnitude of the effect may account for the failure of insulin to reverse dexamethasone enhancement of neurologic injury completely in a previous report that used the rat model of forebrain ischemia. PMID- 11046218 TI - Isoflurane pretreatment ameliorates postischemic neurologic dysfunction and preserves hippocampal Ca2+/calmodulin-dependent protein kinase in a canine cardiac arrest model. AB - BACKGROUND: Inhalational anesthetics are neuroprotective in rat models of global ischemia. To determine whether isoflurane at a clinically relevant concentration is neuroprotective in a canine model of cardiac arrest, we measured neurologic function and hippocampal Ca2+/calmodulin-dependent protein kinase II (CaMKII) content 20 h after cardiac arrest. METHODS: We tested the neuroprotective effect of 30 min of 1.5% isoflurane exposure before 8 min of global ischemia induced with ventricular fibrillation. Animals were randomized to four groups: control, isoflurane-control, ischemia, and isoflurane-ischemia. After resuscitation and 20 h of intensive care, each animal's neurologic deficit score was determined by two blinded evaluators. The hippocampal content of CaMKII, determined by immunoblotting, was measured by an individual blinded to the treatment groups. CaMKII activity was measured in samples from the cortex, hippocampus, and striatum of animals in each group. RESULTS: Isoflurane-ischemic animals had a median neurologic deficit score of 22.6% compared with 43.8% for the ischemic animals (P < 0.05). Hippocampal levels of the beta-subunit of CaMKII (CaMKIIbeta) were relatively preserved in isoflurane-ischemic animals (68 +/- 4% of control) compared with ischemic animals (48 +/- 2% of control; P < 0.001), although both groups were statistically significantly lower than control (P < 0. 001 ischemia vs. control and P < 0.05 isoflurane-ischemia vs. control). CONCLUSIONS: Isoflurane is an effective neuroprotective drug in a canine cardiac arrest model in terms of both functional and biochemical criteria. PMID- 11046219 TI - Alterations in ascending dorsal horn neurons by a surgical incision in the rat foot. AB - BACKGROUND: Little is known about the mechanisms of pain caused by a surgical incision. The authors have developed a rat model of postoperative pain characterized by decreased withdrawal thresholds to punctate mechanical stimuli after plantar incision. The present studies examined the response characteristics of dorsal horn neurons receiving input from the plantar aspect of the foot before and after a plantar incision placed adjacent to the low threshold area of the receptive field (RF). METHODS: Individual dorsal horn neurons from the lumbar enlargement were antidromically identified and characterized as low threshold, wide dynamic range (WDR), and high threshold (HT) based on their responses to brush and pinch. Thresholds (in millinewtons), the pinch RF, and stimulus response functions (SRFs) to von Frey filaments characterized the neurons. SRFs were analyzed using area under the curve. Changes in background activity, punctate mechanical thresholds, SRFs, and RF were recorded after an incision was made adjacent to the most sensitive area of the RF. RESULTS: In all cells, an incision increased background activity; this remained elevated in 3 of 9 HT and 16 of 28 WDR neurons 1 h later. The SRFs were enhanced in 10 of 27 WDR neurons and in 2 of 8 HT cells after incision. Only the WDR neurons were responsive to filaments that produced withdrawal responses after incision in behavioral experiments. Increases in the RFs outside of the injured area occurred after incision in 15 of 29 WDR and 2 of 9 HT cells. CONCLUSION: A plantar incision caused dorsal horn cell activation and central sensitization. Because the threshold of HT neurons did not decrease to the range of the withdrawal responses in behavioral experiments, particular WDR dorsal horn neurons likely contribute to the reduced withdrawal threshold observed in behavioral experiments. Both WDR and HT neurons are capable of transmitting enhanced responses to strong punctate mechanical stimuli after incision. PMID- 11046220 TI - Neuroprotective effects of riluzole and ketamine during transient spinal cord ischemia in the rabbit. AB - BACKGROUND: Massive release of central excitatory neurotransmitters is an important initial step in ischemic neuronal injury, and modification of this process may provide neuroprotection. We studied the protective effects of the voltage-dependent sodium channel antagonist riluzole and the N-methyl-d-aspartate receptor antagonist ketamine on hind limb motor function and histopathologic outcome in an experimental model of spinal cord ischemia. METHODS: Temporary spinal cord ischemia was induced by 29 min of infrarenal balloon occlusion of the aorta in 60 anesthetized New Zealand white rabbits. Animals were randomly assigned to one of four treatment groups (n = 15 each): group C, saline (control); group R, riluzole, 8 mg/kg intravenously; group K, ketamine, 55 mg/kg intravenously; group RK, riluzole and ketamine. After reperfusion, riluzole treatment was continued with intraperitoneal infusions. Normothermia (38 degrees C) was maintained during ischemia, and rectal temperature was assessed before and after intraperitoneal infusions. Neurologic function, according to Tarlov's criteria, was evaluated every 24 h, and infarction volume and the number of eosinophilic neurons and viable motoneurons in the lumbosacral spinal cord was evaluated after 72 h. RESULTS: Neurologic outcome was better in groups R and RK than in groups C and K. All animals in group C (100%) and all animals but one in group K (93%) were paraplegic 72 h after the ischemic insult versus 53% in group R and 67% in group RK (P < 0.01 each). More viable motoneurons were present in groups R and RK than in controls (P < 0.05). CONCLUSIONS: The data indicate that treatment with riluzole can increase the tolerance of spinal cord motoneurons to a period of normothermic ischemia. Intraischemic ketamine did not provide neuroprotection in this model. PMID- 11046221 TI - Altered expression of cardiac myosin isozymes associated with the malignant hyperthermia genotype in swine. AB - BACKGROUND: Anesthetic-induced malignant hyperthermia (MH) in humans and pigs is associated with dramatic alterations in cardiac function. However, it remains controversial as to whether MH-associated cardiac symptoms represent a primary difference of myocardium or a secondary alteration consequent to increases in the hyperthermic stress. Here the authors describe changes in myosin isoform expression in the hearts of MH-susceptible pigs with and without prior exposure to halothane. METHODS: One group of pigs was diagnosed as MH susceptible by halothane challenge and Hal-1843 nucleotide examination. To determine if there is an effect of halothane exposure, another group of pigs was diagnosed by simple MH genotyping without exposure to halothane. After diagnosis and genotyping, animals with and without exposure to halothane were killed to study cardiac myosin isozyme distributions, cardiac myofibrillar adenosine triphosphatase (ATPase) activity, and the steepness of the Ca2+-ATPase activity relation in the hearts of normal and susceptible pigs. The altered myosin isozyme expression was analyzed by pyrophosphate gel electrophoresis. RESULTS: Malignant hyperthermia-susceptible animals with the prior halothane challenge showed an increased V1 myosin (-44%) expression, increased myofibrillar ATPase activity (-25%) and increased steepness of the Ca2+-ATPase activity relation. Without exposure to halothane, no change of myofibrillar ATPase activity was found in the hearts of different genotyped pigs, but there was a small increase in expression of V1 myosin (-5%) in the mutant (TT). CONCLUSIONS: The potential modulation of V1 myosin expression occurs in the hearts of MH-susceptible pigs. The added stress by halothane challenge would further cause a V3 --> V1 shift, which may be attributed to the long-term effects of hyperthermic stress. PMID- 11046222 TI - Pressure-time curve predicts minimally injurious ventilatory strategy in an isolated rat lung model. AB - BACKGROUND: We tested the hypothesis that the pressure-time (P-t) curve during constant flow ventilation can be used to set a noninjurious ventilatory strategy. METHODS: In an isolated, nonperfused, lavaged model of acute lung injury, tidal volume and positive end-expiratory pressure were set to obtain: (1) a straight P t curve (constant compliance, minimal stress); (2) a downward concavity in the P t curve (increasing compliance, low volume stress); and (3) an upward concavity in the P-t curve (decreasing compliance, high volume stress). The P-t curve was fitted to: P = a. tb +c, where b describes the shape of the curve, b = 1 describes a straight P-t curve, b < 1 describes a downward concavity, and b > 1 describes an upward concavity. After 3 h, lungs were analyzed for histologic evidence of pulmonary damage and lavage concentration of inflammatory mediators. Ventilator-induced lung injury occurred when injury score and cytokine concentrations in the ventilated lungs were higher than those in 10 isolated lavaged rats kept statically inflated for 3 h with an airway pressure of 4 cm H2O. RESULTS: The threshold value for coefficient b that discriminated best between lungs with and without histologic and inflammatory evidence of ventilator induced lung injury (receiver-operating characteristic curve) ranged between 0.90 1.10. For such threshold values, the sensitivity of coefficient b to identify noninjurious ventilatory strategy was 1.00. A significant relation (P < 0.001) between values of coefficient b and injury score, interleukin-6, and macrophage inflammatory protein-2 was found. CONCLUSIONS: The predictive power of coefficient b to predict noninjurious ventilatory strategy in a model of acute lung injury is high. PMID- 11046223 TI - Dual effects of intravenous anesthetics on the function of norepinephrine transporters. AB - BACKGROUND: Norepinephrine transporters (NETs) terminate the neuronal transmission of norepinephrine, which is released from noradrenergic neurons. To investigate the interaction with NET, the authors examined the effects of short- and long-term treatment with anesthetics on the activity and mRNA level of NET. METHODS: To assay [3H]norepinephrine uptake, bovine adrenal medullary cells in culture were incubated with [3H]norepinephrine in the presence of intravenous anesthetics, including propofol, thiamylal, and diazepam. To study the direct interaction between the anesthetics and NET, the effect of propofol on the binding of [3H]desipramine to the plasma membrane was examined. To study the long term effect of anesthetics, [3H]norepinephrine uptake by cells pretreated with propofol for 6-24 h and [3H]desipramine binding after pretreatment for 12 h were measured. Simultaneously, we examined the effect of anesthetics on the expression of NET mRNA using the reverse transcriptase-polymerase chain reaction. RESULTS: All of the intravenous anesthetics inhibited [3H]norepinephrine uptake in a concentration-dependent manner. The active concentrations of propofol (1-3 microm) and thiamylal (< or = 30 microm) were similar to those encountered clinically. The kinetic analysis revealed that all the anesthetics noncompetitively inhibited [3H]norepinephrine uptake. Propofol inhibited [3H]desipramine binding with a potency similar to that observed in [3H]norepinephrine uptake. Scatchard analysis showed that propofol competitively inhibited [3H]desipramine binding. On the other hand, long-term treatment of cells with propofol (10 microm) enhanced the NET functional activity and [3H]desipramine binding, and also increased the level of NET mRNA. CONCLUSIONS: These results suggest that intravenous anesthetics have a dual effect on NET; short-term treatment causes inhibition, whereas long-term treatment leads to up regulation. The interaction of intravenous anesthetics with NET may modulate the neuronal transmission of norepinephrine during anesthesia. PMID- 11046224 TI - Development and clinical application of electroencephalographic bispectrum monitoring. PMID- 11046225 TI - Clinical uses of alpha2 -adrenergic agonists. PMID- 11046226 TI - Ventilator alarm failure due to modification of the scavenging system. PMID- 11046227 TI - Acute thrombosis of the external iliac artery after a short procedure in the high lithotomy position. PMID- 11046228 TI - Acute subdural hematoma following spinal anesthesia with a very small spinal needle. PMID- 11046229 TI - Intraoperative hemodialysis during emergency laparotomy. PMID- 11046230 TI - Seizure-like activity on emergence from sevoflurane anesthesia. PMID- 11046231 TI - Ventricular tachycardia caused by hyperkalemia after administration of hypertonic mannitol. PMID- 11046232 TI - More on problems with removing the arrow FlexTip epidural catheter: smooth in hardly out? PMID- 11046233 TI - Rapid opioid detoxification during general anesthesia: is death not a significant outcome? PMID- 11046234 TI - Which parameter measures the effectiveness of volume preload in pregnant patients? PMID- 11046235 TI - Patient in "sniffing position". PMID- 11046236 TI - Ten milligrams intrathecal bupivacaine is too high for spinal anesthesia for hip surgery in the geriatric population. PMID- 11046237 TI - Historical perspective of the "sniffing position". PMID- 11046239 TI - Path integrals for the quantum microcanonical ensemble AB - Path integral representations for the quantum microcanonical ensemble are presented. In the quantum microcanonical ensemble, two operators are of primary interest. First, rhoinsertion mark=delta(E-Hinsertion mark) corresponds to the microcanonical density matrix and can be used to calculate expectation values. Second, Ninsertion mark=Theta(E-Hinsertion mark) can give the number of states with energy E(n) and Theta(x,x('),E)=. A path integral formalism leads to exact integral representations for Omega(x,x('),E) and Theta(x,x('),E). We present both phase space and configuration space forms. For simple systems, such as the free particle and harmonic oscillator, exact solutions are possible. For more complicated systems, expansion schemes or numerical evaluations are required. A perturbative calculation and numerical integration results are presented for the quantum anharmonic oscillator. PMID- 11046238 TI - Epinephrine plasma levels also vary at similar infused doses. PMID- 11046240 TI - Collective modes in an open microwave billiard AB - Numerical calculations for a microwave Sinai billiard coupled strongly to a lead are performed as a function of the coupling strength between billiard and lead. They prove the formation of different time scales in an open quantum system at large coupling strength. The short-lived collective states are formed together with many long-lived trapped states. PMID- 11046241 TI - Optical properties of nonequilibrium low-dimensional systems AB - The optical properties of low-dimensional carrier systems ("quantum wire" type) driven away from equilibrium are studied. The frequency and wave-vector-dependent dielectric function of a quasi-one-dimensional electron system under the action of an exciting external pumping source is derived. The optical responses of the system are obtained in terms of its nonequilibrium thermodynamic state, the latter characterized resorting to a nonequilibrium statistical ensemble formalism. PMID- 11046242 TI - Roughening and preroughening transitions in crystal surfaces with double-height steps AB - We investigate phase transitions in a solid-on-solid model where double-height steps as well as single-height steps are allowed. Without the double-height steps, repulsive interactions between up-up or down-down step pairs give rise to a disordered flat phase. When the double-height steps are allowed, two single height steps can merge into a double-height step (step doubling). We find that the step doubling reduces repulsive interaction strength between single-height steps and that the disordered flat phase is suppressed. As a control parameter a step doubling energy is introduced, which is assigned to each step doubling vertex. From transfer matrix type finite-size-scaling studies of interface free energies, we obtain the phase diagram in the parameter space of the step energy, the interaction energy, and the step doubling energy. PMID- 11046243 TI - Scaling of waves in the bak-tang-wiesenfeld sandpile model AB - We study probability distributions of waves of topplings in the Bak-Tang Wiesenfeld model on hypercubic lattices for dimensions D>/=2. Waves represent relaxation processes which do not contain multiple toppling events. We investigate bulk and boundary waves by means of their correspondence to spanning trees, and by extensive numerical simulations. While the scaling behavior of avalanches is complex and usually not governed by simple scaling laws, we show that the probability distributions for waves display clear power-law asymptotic behavior in perfect agreement with the analytical predictions. Critical exponents are obtained for the distributions of radius, area, and duration of bulk and boundary waves. Relations between them and fractal dimensions of waves are derived. We confirm that the upper critical dimension D(u) of the model is 4, and calculate logarithmic corrections to the scaling behavior of waves in D=4. In addition, we present analytical estimates for bulk avalanches in dimensions D>/=4 and simulation data for avalanches in DJ2, with equal probability. Using an iterative method, based on a successive application of the star-triangle transformation, we have determined at the bulk critical temperature the correlation length along the strip xi(L) for different widths of the strip L2. The eigenvalue density in the complex plane is nonzero within a wedge that encloses the negative real axis. Particle motion is diffusive at long times, but for short times we find a novel time dependence of the mean-square displacement, approximately t(2/d) in dimension d>2, associated with the imaginary parts of eigenvalues. PMID- 11046256 TI - Moment analysis of the probability distribution of different sandpile models AB - We reconsider the moment analysis of the Bak-Tang-Wiesenfeld and the stochastic sandpile model introduced by Manna [J. Phys. A 24, L363 (1991)] in two and three dimensions. In contrast to recently performed investigations our analysis reveals that the models are characterized by different scaling behavior, i.e., they belong to different universality classes. PMID- 11046257 TI - Solvable kinetic gaussian model in an external field AB - In this paper, the single-spin transition dynamics is used to investigate the kinetic Gaussian model in a periodic external field. We first derive the fundamental dynamic equations, and then treat an isotropic d-dimensional hypercubic lattice Gaussian spin system with Fourier's transformation method. We obtain exactly the local magnetization and the equal-time pair-correlation function. The critical characteristics of the dynamical relaxation tau(q), the complex susceptibility chi(omega,q), and the dynamical response are discussed. The results show that the time evolution of the dynamical quantities and the dynamical responses of the system strongly depend on the frequency and the wave vector of the external field. PMID- 11046258 TI - Mixed-spin ising model with one- and two-spin competing dynamics AB - In this work we found the stationary states of a kinetic Ising model, with two different types of spins: sigma=1/2 and S=1. We divided the spins into two interpenetrating sublattices, and found the time evolution for the probability of the states of the system. We employed two transition rates which compete between themselves: one, associated with the Glauber process, which describes the relaxation of the system through one-spin flips; the other, related to the simultaneous flipping of pairs of neighboring spins, simulates an input of energy into the system. Using the dynamical pair approximation, we determined the equations of motion for the sublattice magnetizations, and also for the correlation function between first neighbors. We found the phase diagram for the stationary states of the model, and we showed that it exhibits two continuous transition lines: one line between the ferrimagnetic and paramagnetic phases, and the other between the paramagnetic and antiferrimagnetic phases. PMID- 11046259 TI - Nonequilibrium phase transitions induced by multiplicative noise: effects of self correlation AB - A recently introduced lattice model, describing an extended system which exhibits a reentrant (symmetry-breaking, second-order) noise-induced nonequilibrium phase transition, is studied under the assumption that the multiplicative noise leading to the transition is colored. Within an effective Markovian approximation and a mean-field scheme it is found that when the self-correlation time tau of the noise is different from zero, the transition is also reentrant with respect to the spatial coupling D. In other words, at variance with what one expects for equilibrium phase transitions, a large enough value of D favors disorder. Moreover, except for a small region in the parameter subspace determined by the noise intensity sigma and D, an increase in tau usually prevents the formation of an ordered state. These effects are supported by numerical simulations. PMID- 11046260 TI - Stochastic resonance as dithering AB - A direct correspondence is demonstrated between the phenomenon of "stochastic resonance" in static nonlinear systems and the dithering effect well known in the theory of digital waveform coding. It is argued that many static systems displaying stochastic resonance are forms of dithered quantizers, and that the existence or absence of stochastic resonance in such systems can be predicted from the effects of "dither averaging" upon their transfer characteristics. Also, results are introduced regarding stochastic resonance in certain nonlinear systems with memory (e.g., hysteretic systems). PMID- 11046261 TI - Event-by-event fluid dynamcis AB - Coarse grained Langevin-type effective field equations may provide some guidance for the analysis of mesoscopic or microscopic molecular systems exhibiting fluctuations, or for systems of hundreds to thousands of atomic or subatomic particles produced in atomic or high-energy nuclear collisions. Suggestions for consistent realization of random fluctuations in discretized fluid dynamics will be presented. PMID- 11046262 TI - Effect of shear on phase-ordering dynamics with order-parameter-dependent mobility: the large-n limit AB - The effect of shear on the ordering kinetics of a conserved order-parameter system with O(n) symmetry and order-parameter-dependent mobility Gamma(straight phi-->) approximately (1-straight phi-->(2)/n)(alpha) is studied analytically within the large-n limit. In the late stage, the structure factor becomes anisotropic and exhibits multiscaling behavior with characteristic length scales (t(2alpha+5)/ln t)(1/2(alpha+2)) in the flow direction and (t/ln t)(1/2(alpha+2)) in directions perpendicular to the flow. As in the alpha=0 case, the structure factor in the shear-flow plane has two parallel ridges. PMID- 11046263 TI - Universality classes for the ricepile model with absorbing properties AB - The absorbing "ricepile" model with stochastic toppling rules has been numerically studied. Local limited, local unlimited, nonlocal limited, and nonlocal unlimited versions of the absorbing model have been investigated. Transport properties and different dynamical regimes of all of the models have been analyzed, from the point of view of self-organized criticality. Phase transitions between different dynamical regimes were studied in detail. It was shown that the absorbing models belong to two different universality classes. PMID- 11046264 TI - Distribution of escape times for a deterministically driven bistable system AB - In this paper, we analyze the sequence of escape times for a particle in a symmetric double-well potential coupled to a chain of monodimensional oscillators and we find that, in some range of energies, the probability of escape exhibits the multimodal form that is characteristic of bistable systems driven by a periodic signal embedded in noise. We identify two different modes contributing to the overall hopping dynamics of the particle, each one having a definite dependence on the energy of the chain. Those findings suggest a model for internal fluctuations that could be useful in the study of some problems of interest in physics and biology. PMID- 11046265 TI - Thermodynamic picture of the glassy state gained from exactly solvable models AB - A picture for thermodynamics of the glassy state was introduced recently by us [Phys. Rev. Lett. 79, 1317 (1997); 80, 5580 (1998)]. It starts by assuming that one extra parameter, the effective temperature, is needed to describe the glassy state. This approach connects responses of macroscopic observables to a field change with their temporal fluctuations, and with the fluctuation-dissipation relation, in a generalized, nonequilibrium way. Similar universal relations do not hold between energy fluctuations and the specific heat. In the present paper, the underlying arguments are discussed in greater length. The main part of the paper involves details of the exact dynamical solution of two simple models introduced recently: uncoupled harmonic oscillators subject to parallel Monte Carlo dynamics, and independent spherical spins in a random field with such dynamics. At low temperature, the relaxation time of both models diverges as an Arrhenius law, which causes glassy behavior in typical situations. In the glassy regime, we are able to verify the above-mentioned relations for the thermodynamics of the glassy state. In the course of the analysis, it is argued that stretched exponential behavior is not a fundamental property of the glassy state, though it may be useful for fitting in a limited parameter regime. PMID- 11046266 TI - Boundary spatiotemporal correlations in a self-organized critical model of punctuated equilibrium AB - In a semi-infinite geometry, a one-dimensional, M-component model of biological evolution realizes microscopically an inhomogeneous branching process for M- >infinity. This implies a size distribution exponent tau(')=7/4 for avalanches starting at a free, "dissipative" end of the evolutionary chain. A bulklike behavior with tau(')=3/2 is restored by "conservative" boundary conditions. These are such as to strictly fix to its critical, bulk value the average number of species directly involved in an evolutionary avalanche by the mutating species located at the chain end. A two-site correlation function exponent tau(')(R)=4 is also calculated exactly in the "dissipative" case, when one of the points is at the border. Together with accurate numerical determinations of the time recurrence exponent tau(')(first), these results show also that, no matter whether dissipation is present or not, boundary avalanches have the same space and time fractal dimensions as those in the bulk, and their distribution exponents obey the basic scaling laws holding there. PMID- 11046267 TI - Dynamical weight functions for a planar crack AB - The stress intensity factors are evaluated for a moving planar crack for loadings which vary arbitrarily in time and three dimensions of space. We exploit the adjoint elasticity equation obeyed by the corresponding weight functions, and a new and more universal Wiener-Hopf factorization of the Rayleigh function, this being the central difficulty in such calculations. For the mode II weight function we give further asymptotic results crucial to a subsequent calculation of crack stability with respect to out-of-plane perturbations. PMID- 11046268 TI - Drift ratchet AB - We consider a silicon wafer, pierced by millions of identical pores with periodically varying diameters but without spatial inversion symmetry (ratchet profile). When a liquid is periodically pumped back and forth through the pores, our theory predicts a net transport of suspended micrometer-sized particles (drift ratchet). The direction of this particle current depends very sensitively on the size of the particles. For typical parameter values of the experiment, two different types of particles at an initially homogeneous 1:1 mixture are spatially separated with a purity beyond 1:1000 on a time scale of a few hours in comparably large quantities. This result is due to the highly parallel architecture of the device. The experimental realization of the setup, presently under construction, thus appears to be a promising new particle separation device, possibly superior to existing methods for particles sizes on the micrometer scale. PMID- 11046269 TI - Diffusion in disordered lattices and related heisenberg ferromagnets AB - We study the diffusion of classical hard-core particles in disordered lattices within the formalism of a quantum spin representation. This analogy enables an exact treatment of noninstantaneous correlation functions at finite particle densities in terms of single spin excitations in disordered ferromagnetic backgrounds. Applications to diluted chains and percolation clusters are discussed. It is found that density fluctuations in the former exhibit a stretched exponential decay while an anomalous power law asymptotic decay is conjectured for the latter. PMID- 11046270 TI - Scaling of lyapunov exponents of coupled chaotic systems AB - We develop a statistical theory of the coupling sensitivity of chaos. The effect was first described by Daido [Prog. Theor. Phys. 72, 853 (1984)]; it appears as a logarithmic singularity in the Lyapunov exponent in coupled chaotic systems at very small couplings. Using a continuous-time stochastic model for the coupled systems we derive a scaling relation for the largest Lyapunov exponent. The singularity is shown to depend on the coupling and the systems' mismatch. Generalizations to the cases of asymmetrical coupling and three interacting oscillators are considered, too. The analytical results are confirmed by numerical simulations. PMID- 11046271 TI - Homoclinic tangency and chaotic attractor disappearance in a dripping faucet experiment AB - A sequence of attractors, reconstructed from interdrops time series data of a leaky faucet experiment, is analyzed as a function of the mean dripping rate. We established the presence of a saddle point and its manifolds in the attractors and we explained the dynamical changes in the system using the evolution of the manifolds of the saddle point, as suggested by the orbits traced in first return maps. The sequence starts at a fixed point and evolves to an invariant torus of increasing diameter (a Hopf bifurcation) that pushes the unstable manifold towards the stable one. The torus breaks up and the system shows a chaotic attractor bounded by the unstable manifold of the saddle. With the attractor expansion the unstable manifold becomes tangential to the stable one, giving rise to the sudden disappearance of the chaotic attractor, which is an experimental observation of a so called chaotic blue sky catastrophe. PMID- 11046272 TI - Dynamical control of systems near bifurcation points using time series AB - In order to excite experimentalists to apply a dynamical control method [Zhao et al., Phys. Rev. E 53, 299 (1996); 57, 5358 (1998)], we further introduce a simplified control law in this paper. The law provides a convenient way (in certain circumstance a necessary way) for experimentalists to achieve the system control when the exact position of the desired control objective cannot be known in advance. The validity of the control law is rigidly verified when the system nears a bifurcation point but our numerical examples show that it can be extended to a wide parameter region practically. PMID- 11046273 TI - Dynamical system approach to phyllotaxis AB - This paper presents a bifurcation study of a model widely used to discuss phyllotactic patterns, i.e., leaf arrangements. Although stable patterns can be easily obtained by numerical simulations, a stability or bifurcation analysis is hindered by the fact that the model is defined by an algorithm and not a dynamical system, mainly because new active elements are added at each step, and thus the dimension of the "natural" phase space is not conserved. Here a construction is presented by which a well defined dynamical system can be obtained, and a bifurcation analysis can be carried out. Stable and unstable patterns are found by an analytical relation, in which the roles of different growth mechanisms determining the shape is clarified. Then bifurcations are studied, especially anomalous scenarios due to discontinuities embedded in the original model. Finally, an explicit formula for evaluation of the Jacobian, and thus the eigenvalues, is given. It is likely that problems of the above type often arise in biology, and especially in morphogenesis, where growing systems are modeled. PMID- 11046274 TI - Autocorrelation function of level velocities for ray-splitting billiards AB - We study experimentally and theoretically the autocorrelation function of level velocities c(x) and the generalized conductance C(0) for classically chaotic ray splitting systems. Experimentally, a Sinai ray-splitting billiard was simulated by a thin microwave rectangular cavity with a quarter-circle Teflon insert. For the theoretical estimates of the autocorrelator c(x) and the conductance C(0) we made parameter-dependent quantum calculations of eigenenergies of an annular ray splitting billiard. Our experimental and numerical results are compared to theoretical predictions of systems based on the Gaussian orthogonal ensemble in random matrix theory. PMID- 11046275 TI - Synchronization in a system of globally coupled oscillators with time delay AB - We study the synchronization phenomena in a system of globally coupled oscillators with time delay in the coupling. The self-consistency equations for the order parameter are derived, which depend explicitly on the amount of delay. Analysis of these equations reveals that the system in general exhibits discontinuous transitions in addition to the usual continuous transition, between the incoherent state and a multitude of coherent states with different synchronization frequencies. In particular, the phase diagram is obtained on the plane of the coupling strength and the delay time, and ubiquity of multistability as well as suppression of the synchronization frequency is manifested. Numerical simulations are also performed to give consistent results. PMID- 11046276 TI - Quantum chaotic scattering with a mixed phase space: the three-disk billiard in a magnetic field AB - We study the classical and semiclassical scattering behavior of electrons in an open three-disk billard in the presence of a homogeneous magnetic field, which is confined to the inner part of the scattering region. As the magnetic field is increased the phase space of the invariant set of the classical scattering trajectories changes from hyperbolic (fully chaotic) to a mixed situation, where KAM tori are present. The "stickiness" of the stable trajectories leads to a much slower decay of the survival probability of trajectories as compared to the hyperbolic case. We show that this effect influences strongly the quantum fluctuations of the scattering amplitude and cross sections. PMID- 11046277 TI - Stability analysis of plane wave solutions of the discrete ginzburg-landau equation AB - The discrete Ginzburg-Landau model for a family of oscillators linearly coupled with their first neighbors is studied. The full linear stability analysis of the nonlinear plane wave solutions is performed by considering both the wave number (k) of the basic states and the wave number (q) of the perturbations as free parameters. In particular, it is shown that nonlinear plane waves can be destabilized not only by long (q-->0) or short (q=pi) wave perturbations, but also by intermediate wave numbers (0(n+1) square that occur when the character of the constrained space evolves from being two dimensional to being three dimensional (triangle up denotes layers with hexagonal packing symmetry, while square denotes layers with square packing symmetry). The two transitions, ntriangle up-->n buckled-->(n+1) square, are found to be first order. PMID- 11046310 TI - Hexagonal to square lattice conversion in bilayer systems AB - We report the results of extensive molecular dynamics simulations of the reconstructive hexagonal to square lattice conversion in bilayer colloid systems. Two types of interparticle potential were used to represent the colloid-colloid interactions in the suspension. One potential, due to Marcus and Rice, is designed to describe the interaction of sterically stabilized colloid particles. This potential has a term that represents the attraction between colloid particles when there is incipient overlap between the stabilizing brushes on their surfaces, a (soft repulsion) term that represents the entropy cost associated with interpenetration of the stabilizing brushes, and a term that represents core-core repulsion. The other potential we used is an almost hard core repulsion with continuous derivatives. Our results clearly show that the character of the reconstructive hexagonal to square lattice conversion in bilayer colloid systems is potential dependent. For a system with colloid-colloid interactions of the Marcus-Rice type, the packing of particles in the square array exhibits a large interlayer lattice spacing, with the particles located at the minima of the attractive well. In this case the hexagonal to square lattice transition is first order. For a system with hard core colloid-colloid interactions there are two degenerate stable intermediate phases, linear and zigzag rhombic, that are separated from the square lattice by strong first order transitions, and from the hexagonal lattice by either weak first or second order transitions. PMID- 11046311 TI - Adiabatic compressibility of AOT AB - The self-assembly of amphiphilic molecules into supramolecular aggregates involves a number of complex phenomena and forces. Recent developments of highly sensitive, densimetric and acoustic methods on small volume samples have provided novel sensitive probes to explore the physical properties of these complex fluids. We have investigated, by high precision densimetry and ultrasound velocimetry, reverse micelles of [sodium bis(2-ethylhexyl)sulfosuccinate] in oil (isooctane and decane), at increasing water concentration and at variable micellar volume fractions. The size of these spherical micelles has been determined by small angle x-ray scattering. Using these results, in the framework of the effective medium theory, we have developed a simple model of micellar compressibility, allowing the calculation of physical parameters (aggregation number, volume, and compressibility) of the surfactant monomolecular film as well as that of the micellar waters. In particular, we show that the central aqueous core designated as "free" water, located at a distance from the oil-water interacting interface, is twice as compressible as "bulk" water. One notable feature of this work is the influence of the nature of the oil on the above parameters. PMID- 11046312 TI - Effect of the wall roughness on slip and rheological properties of hexadecane in molecular dynamics simulation of couette shear flow between two sinusoidal walls AB - Molecularly thin liquid films of alkanes in extreme conditions in a boundary lubrication regime have been investigated. The wall is modeled as a rough atomic sinusoidal wall. The effect on the boundary condition of the roughness characteristics, given by the period and amplitude of the sinusoidal wall, is studied here. The effect of the molecular length of the lubricating fluid is also examined here. The results show that the relative size of the fluid molecules and wall roughness determines the slip or nonslip boundary conditions. The effect of wall roughness characteristics on the rheological properties of the lubrication film is also studied. PMID- 11046313 TI - Instabilities of concentration stripe patterns in ferrocolloids AB - Equations describing the kinetics of the phase separation in ferrocolloids in a Hele-Shaw cell under the action of a rotating magnetic field are proposed. Numerical simulation on the basis of a pseudospectral technique demonstrates that upon the action of a rotating field on a magnetic colloid which undergoes the phase separation a periodical system of stripes parallel to the plane of a rotating magnetic field stripes is created. The period of a structure found numerically satisfactorily corresponds to the one calculated on the basis of the energy minimum. Thus, the undulation instability leading to the formation of chevron structures takes place if the tangential component of a rotating magnetic field is eliminated, whereas the normal component is increased at the same time. If during the development of the undulation deformations of a concentration pattern the magnetic Bond number is large enough the secondary instabilities may occur leading to the fingering of stripes to bring about merging and break-up of stripes. It is shown that an increase in the magnetic Bond number leads to the onset of the instability at the boundaries between the regions with homogeneous orientation of stripes as well as to formation of the characteristic hairpin patterns. PMID- 11046314 TI - Diffusional mechanism of strong selection in ostwald ripening AB - The purpose of this paper is to show through a systematic asymptotic analysis that fluctuations, accounted for as a diffusional perturbation in the Lifshitz Slyozov-Wagner (LSW) model of Ostwald ripening, provides, as conjectured previously by Meerson [Phys. Rev. E 60, 3072 (1999)], a "strong" selection of the limiting solution, out of a one-parameter family of similarity solutions with a finite support, as the sole attractor of time evolution. Throughout the latter, the previously described weak selection of other similarity solutions of that family, by the initial conditions with finite supports, occurs as intermediate time asymptotics. The respective mechanism is traced first for a simple instance of the LSW model with linear characteristic equations (integer power in the particle growth rate law equals -1), beginning with the analysis of steady states in the perturbed problem in similarity variables and weak selection in the unperturbed problem, followed by a detailed asymptotic analysis of the time dependent perturbed problem, and generalized next for an arbitrary integer power in the range [-1,2]. The approximate asymptotic solutions obtained are compared with the exact numerical ones. PMID- 11046315 TI - Responses of a Hodgkin-Huxley neuron to various types of spike-train inputs. AB - Numerical investigations have been made of responses of a Hodgkin-Huxley (HH) neuron to spike-train inputs whose interspike interval (ISI) is modulated by deterministic, semi-deterministic (chaotic), and stochastic signals. As deterministic one, we adopt inputs with the time-independent ISI and with time dependent ISI modulated by sinusoidal signal. The Rossler and Lorentz models are adopted for chaotic modulations of ISI. Stochastic ISI inputs with the gamma distribution are employed. It is shown that distribution of output ISI data depends not only on the mean of ISIs of spike-train inputs but also on their fluctuations. The distinction of responses to the three kinds of inputs can be made by return maps of input and output ISIs, but not by their histograms. The relation between the variations of input and output ISIs is shown to be different from that of the integrate and fire (IF) model because of the refractory period in the HH neuron. PMID- 11046316 TI - From local to global spatiotemporal chaos in a cardiac tissue model. AB - Two kinds of chaos can occur in cardiac tissue, chaotic meander of a single intact spiral wave and chaotic spiral wave breakup. We studied these behaviors in a model of two-dimensional cardiac tissue based on the Luo-Rudy I action potential model. In the chaotic meander regime, chaos is spatially localized to the core of the spiral wave. When persistent spiral wave breakup occurs, there is a transition from local to global spatiotemporal chaos. PMID- 11046317 TI - Short-term forecasting of life-threatening cardiac arrhythmias based on symbolic dynamics and finite-time growth rates. AB - Ventricular tachycardia or fibrillation (VT-VF) as fatal cardiac arrhythmias are the main factors triggering sudden cardiac death. The objective of this study is to find early signs of sustained VT-VF in patients with an implanted cardioverter defibrillator (ICD). These devices are able to safeguard patients by returning their hearts to a normal rhythm via strong defibrillatory shocks; additionally, they store the 1000 beat-to-beat intervals immediately before the onset of a life threatening arrhythmia. We study these 1000 beat-to-beat intervals of 17 chronic heart failure ICD patients before the onset of a life-threatening arrhythmia and at a control time, i.e., without a VT-VF event. To characterize these rather short data sets, we calculate heart rate variability parameters from the time and frequency domain, from symbolic dynamics as well as the finite-time growth rates. We find that neither the time nor the frequency domain parameters show significant differences between the VT-VF and the control time series. However, two parameters from symbolic dynamics as well as the finite-time growth rates discriminate significantly both groups. These findings could be of importance in algorithms for next generation ICD's to improve the diagnostics and therapy of VT VF. PMID- 11046318 TI - Coherence resonance and noise-induced synchronization in globally coupled Hodgkin Huxley neurons. AB - The coherence resonance (CR) of globally coupled Hodgkin-Huxley neurons is studied. When the neurons are set in the subthreshold regime near the firing threshold, the additive noise induces limit cycles. The coherence of the system is optimized by the noise. The coupling of the network can enhance CR in two different ways. In particular, when the coupling is strong enough, the synchronization of the system is induced and optimized by the noise. This synchronization leads to a high and wide plateau in the local CR curve. A bell shaped curve is found for the peak height of power spectra of the spike train, being significantly different from a monotonic behavior for the single neuron. The local-noise-induced limit cycle can evolve to a refined spatiotemporal order through the dynamical optimization among the autonomous oscillation of an individual neuron, the coupling of the network, and the local noise. PMID- 11046319 TI - Elastic stability of DNA configurations. I. General theory. AB - Results are presented in the theory of the elastic rod model for DNA, among which are criteria enabling one to determine whether a calculated equilibrium configuration of a DNA segment is stable in the sense that it gives a local minimum to the sum of the segment's elastic energy and the potential of forces acting on it. The derived stability criteria are applicable to plasmids and to linear segments subject to strong anchoring end conditions. Their utility is illustrated with an example from the theory of configurations of the extranucleosomal loop of a DNA miniplasmid in a mononucleosome, with emphasis placed on the influence that nicking and ligation on one hand, and changes in the ratio of elastic coefficients on the other, have on the stability of equilibrium configurations. In that example, the configurations studied are calculated using an extension of the method of explicit solutions to cases in which the elastic rod modeling a DNA segment is considered impenetrable, and hence excluded volume effects and forces arising from self-contact are taken into account. PMID- 11046321 TI - Different hierarchy of avalanches observed in the bak-sneppen evolution model AB - A quantity &fmacr; denoting the average fitness of an ecosystem is introduced in the Bak-Sneppen model. Through this quantity, a different hierarchy of avalanches, &fmacr;(0) avalanche, is observed in the evolution of Bak-Sneppen model. An exact gap equation, governing the self-organization of the model, is presented in terms of &fmacr;. It is found that self-organized threshold &fmacr;(c) can be exactly obtained. Two basic exponents of the new avalanche tau, avalanche distribution, and D, avalanche dimension are given through simulations of one- and two-dimensional Bak-Sneppen models. It is suggested that &fmacr; may be a good quantity in determining the emergence of criticality. PMID- 11046320 TI - Elastic stability of DNA configurations. II. Supercoiled plasmids with self contact. AB - Configurations of protein-free DNA miniplasmids are calculated with the effects of impenetrability and self-contact forces taken into account by using exact solutions of Kirchhoff's equations of equilibrium for elastic rods of circular cross section. Bifurcation diagrams are presented as graphs of excess link, DeltaL, versus writhe, W, and the stability criteria derived in paper I of this series are employed in a search for regions of such diagrams that correspond to configurations that are stable, in the sense that they give local minima to elastic energy. Primary bifurcation branches that originate at circular configurations are composed of configurations with D(m) symmetry (m=2,3,...). Among the results obtained are the following. (i) There are configurations with C2 symmetry forming secondary bifurcation branches which emerge from the primary branch with m=3, and bifurcation of such secondary branches gives rise to tertiary branches of configurations without symmetry. (ii) Whether or not self contact occurs, a noncircular configuration in the primary branch with m=2, called branch alpha, is stable when for it the derivative dDeltaL/dW, computed along that branch, is strictly positive. (iii) For configurations not in alpha, the condition dDeltaL/dW>0 is not sufficient for stability; in fact, each nonplanar contact-free configuration that is in a branch other than alpha is unstable. A rule relating the number of points of self-contact and the occurrence of intervals of such contact to the magnitude of DeltaL, which in paper I was found to hold for segments of DNA subject to strong anchoring end conditions, is here observed to hold for computed configurations of protein-free miniplasmids. PMID- 11046322 TI - Chaotic behavior in noninteger-order cellular neural networks AB - In this paper, a simple system showing chaotic behavior is introduced. It is based on the well-known concept of cellular neural networks (CNNs), which have already given good results in generating complex dynamics. The peculiarity of the CNN model consists in the fact that it replaces the traditional first-order cell with a noninteger-order one. The introduction of the fractional cell, with a suitable choice of the coupling parameters, leads to the onset of chaos in a simple two-cell system. A theoretical approach, based on the harmonic balance theory, has been used to investigate the existence of chaos. A circuit realization of the proposed fractional two-cell chaotic CNN is reported and the corresponding strange attractor is also shown. PMID- 11046323 TI - Surface waves in strongly irradiated dusty plasmas AB - High-frequency surface waves at the interface between two dusty plasmas subject to radiation are considered. Ultraviolet radiation with energy flux larger than the photoelectric work function of the dust surface causes photoemission of electrons. The dust charge and the overall charge balance of the plasma are thus modified. The dispersion properties of the surface waves are investigated for three parameter regimes distinguished by the charging mechanisms in the two plasmas. It is shown that photoemission can significantly affect the plasma and the surface waves. PMID- 11046324 TI - Transverse modulation of an electron beam generated in self-modulated laser wakefield accelerator experiments AB - Low energy electron beams (E approximately 300 keV) generated in a self-modulated laser wakefield accelerator experiment were observed to filament and be deflected away from the laser axis forming radial jets in the electron beam profile. At higher energies (E>900 keV), the filamentation and jets were suppressed and smooth electron beams copropagating with the laser were observed. The observed electron beam filamentation likely results from laser beam filamentation in the plasma due to relativistic self-focusing effects. The radial jets of low energy electrons are likely caused by transverse ejection of the electrons due to the radial structure of the wakefield and space charge deflection of electrons as they exit the laser focus. PMID- 11046325 TI - Dipolar interaction in a colloidal plasma AB - The dipole-dipole interaction between macroscopic grains in a plasma is calculated. The presence of a plasma medium shields the Coulomb grain interactions as well as necessitates that nonzero temperature effects be accounted for. The resulting electrostatic but not so much the Helmholtz free energy shows overshooting when equal strength parallel dipoles align with or are perpendicular to the line joining their centers. More striking is the appearance of a short range attractive well interaction for angles near pi/3. The contribution of the dipole-dipole to the total interaction in so-called plasma crystals has become increasingly recognized. PMID- 11046326 TI - Analysis of the M-shell spectra emitted by a short-pulse laser-created tantalum plasma AB - The spectrum of tantalum emitted by a subpicosecond laser-created plasma, was recorded in the regions of the 3d-5f, 3d-4f, and 3d-4p transitions. The main difference with a nanosecond laser-created plasma spectrum is a broad understructure appearing under the 3d-5f transitions. An interpretation of this feature as a density effect is proposed. The supertransition array model is used for interpreting the spectrum, assuming local thermodynamic equilibrium (LTE) at some effective temperature. An interpretation of the 3d-4f spectrum using the more detailed unresolved transition array formalism, which does not assume LTE, is also proposed. Fitted contributions of the different ionic species differ slightly from the LTE-predicted values. PMID- 11046327 TI - Nonlinear dust kinetic Alfven waves AB - Localized nonlinear dust kinetic Alfven waves are investigated. It is found that finite density dips and humps can coexist. The density humps are cusped and narrower than the dips. PMID- 11046328 TI - Proper orthogonal decomposition and galerkin projection for a three-dimensional plasma dynamical system AB - A general method by which to investigate nonlinear dynamical systems close to a stability threshold is presented. This method combines a proper orthogonal decomposition and a subsequent Galerkin projection. This technique is applied to three-dimensional resistive ballooning plasma fluctuations in a tokamak. The corresponding dynamical system belongs to a large family of convective fluid systems including Rayleigh-Benard convection. A proper orthogonal decomposition of the fluctuating signal obtained by numerical simulation shows that the relevant modes are close to the linear (global) modes. The Galerkin projection provides a low-dimensional system that allows the study of shear flow generation, its subsequent fluctuation reduction, and the evolution to oscillating states. PMID- 11046329 TI - Force between charged particles with ion condensation AB - We have numerically calculated the interaction forces between two highly charged spherical particles embedded in a cloud of small ions with or without charge redistribution on the particle surfaces. Ion condensation near the charged particles leads to reduced electrostatic interaction between the particles, and we find that the effective two-particle interaction is significantly smaller than the values expected from considering only effective single-particle potentials. PMID- 11046330 TI - Hamiltonian dynamics of vortex and magnetic lines in hydrodynamic type systems AB - Vortex line and magnetic line representations are introduced for a description of flows in ideal hydrodynamics and magnetohydrodynamics (MHD), respectively. For incompressible fluids, it is shown with the help of this transformation that the equations of motion for vorticity Omega and magnetic field follow from a variational principle. By means of this representation, it is possible to integrate the hydrodynamic type system with the Hamiltonian H=integral|Omega|dr and some other systems. It is also demonstrated that these representations allow one to remove from the noncanonical Poisson brackets, defined in the space of divergence-free vector fields, the degeneracy connected with the vorticity frozenness for the Euler equation and with magnetic field frozenness for ideal MHD. For MHD, a new Weber-type transformation is found. It is shown how this transformation can be obtained from the two-fluid model when electrons and ions can be considered as two independent fluids. The Weber-type transformation for ideal MHD gives the whole Lagrangian vector invariant. When this invariant is absent, this transformation coincides with the Clebsch representation analog introduced by V.E. Zakharov and E. A. Kuznetsov [Dokl. Ajad. Nauk 194, 1288 (1970) [Sov. Phys. Dokl. 15, 913 (1971)]]. PMID- 11046331 TI - Two-dimensional model of thermal smoothing of laser imprint in a double-pulse plasma AB - The laser prepulse effect on the thermal smoothing of nonuniformities of target illumination is studied by means of a two-dimensional Lagrangian hydrodynamics simulation, based on the parameters of a real experiment. A substantial smoothing effect is demonstrated for the case of an optimum delay between the prepulse and the main heating laser pulse. The enhancement of the thermal smoothing effect by the laser prepulse is caused by the formation of a long hot layer between the region of laser absorption and the ablation surface. A comparison with experimental results is presented. PMID- 11046332 TI - Diffusion cooling in a magnetic field AB - Diffusion cooling of electrons in a weakly ionized plasma in the presence of a magnetic field is studied using the balance equations of momentum transfer theory, well known in "swarm" or test particle analysis. It is shown that for a cylindrical, axially symmetric system, the electron temperature profile can be "hollow" (i.e., T(e)1 for the noncritical one. These values are much smaller than theoretically predicted y=1.8-1.9. Based on the assumption that a strong electric field induces in the mixture a quasinematic structure with semiclassical critical properties, a quantitative explanation of this difference is proposed. PMID- 11046352 TI - Characterization of soliton damping in the granular chain under gravity AB - A soliton created in the horizontal granular chain damps due to gravity in the vertical chain. We show that there are two types of propagating modes, quasisolitary and oscillatory, in the vertical chain, depending on the strength of impulse. We find that the type of damping is a power law in depth or time. We also find that the absolute value of the exponent of the power law decreases as the strength of the initial impulse increases in the quasisolitary regime. In the oscillatory regime, however, in which the initial impulse is weak, the power-law exponent is independent of the strength of the initial impulse. We show that the power-law damping is caused by the gravitation which results in the change of the force constant at each contact. PMID- 11046353 TI - Hysteresis studies in a noisy autoassociative neural network AB - We define magnetization for a noisy autoassociative neural network driven by an external periodic field. Numerical simulations are carried out to investigate the effect of drive amplitude and frequency and noise strength on the area of the hysteresis loop. We observe that in the presence of weak periodic signal, the network exhibits a maximum in the hysteresis loop area at a nonzero noise intensity indicating maximum synchronization between the periodic signal and the response. It also goes through a maximum as a function of signal frequency. PMID- 11046354 TI - Stochastic electron gas theory of coherence in laser-driven synchrotron radiation AB - The transition from coherent to incoherent laser-driven synchrotron radiation is studied within the framework of a stochastic electron gas model. The fundamental difference between this approach and a relativistic fluid model resides in the fact that, for any number of incoherently phased point electrons, the 4-current contains Fourier components at arbitrarily short wavelengths, whereas the fluid model introduces an unphysical cutoff scale. PMID- 11046355 TI - Nonlinear short-wave propagation in ferrites AB - In this paper we discuss the propagation of nonlinear electromagnetic short waves in ferromagnetic insulators. We show that such propagation is perpendicular to an externally applied field. In the nonlinear regime we determine various possible propagation patterns: an isolated pulse, a modulated sinusoidal wave, and an asymptotic two-peak wave. The mathematical structure underlying the existence of these solutions is that of the integrable sine-Gordon equation. PMID- 11046356 TI - Comment on "Monte carlo study of structural ordering in charged colloids using a long-range attractive interaction" AB - A recent theoretical analysis [B. V. R. Tata and N. Ise, Phys. Rev. E 58, 2237 (1998)] of interactions and phase transitions in charge-stabilized colloidal suspensions made reference to our previously published measurements [J. C. Crocker and D. G. Grier, Phys. Rev. Lett. 73, 352 (1994); 77, 1897 (1996); A. E. Larson and D. G. Grier, Nature (London) 385, 230 (1997)] of colloidal interactions. Tata and Ise claim that our measurements cannot distinguish between predictions of the Derjaguin, Landau, Verwey, and Overbeek (DLVO) theory and those of the competing theory due to Sogami and Ise (SI). We demonstrate that the DLVO theory accurately describes the measured interactions between isolated pairs of like-charged spheres, while the SI theory fails both quantitatively and qualitatively to describe our data. PMID- 11046357 TI - Reply to "Comment on 'Monte carlo study of structural ordering in charged colloids using a long-range attractive interaction' " AB - In the Comment by Grier and Crocker (preceding paper) the authors tried to refute our criticism [Phys. Rev. E 58, 2237 (1998)] on their work [J. C. Crocker and D. G. Grier, Phys. Rev. Lett. 77, 1897 (1996)] by simply fitting once again their old experimental data. Grier and Crocker claim that their pair-potential measurements on aqueous dilute suspension of charged colloidal particles confined between charged glass walls at gap of about 8 &mgr;m provide evidence for the failure of Sogami-Ise (SI) theory and demonstrate the applicability of the Dejaguin, Landau, Vervey, and Overbeek (DLVO) theory. Grier and Crocker do not provide additional experimental proof to counter our criticism. We continue to claim here based on our conductivity and conductometric titration measurements, which allow estimating the effective charge and determining the number and nature of the dissociable sites respectively, that their measurements using not well characterized samples cannot provide clear evidence for the failure of SI theory. With the evidences available in literature, we refute all of the Grier and Crocker comments, including the effect of charged wall confinement on the measured colloidal interactions. PMID- 11046358 TI - Universal behavior of nonequilibrium fluctuations in free diffusion processes AB - We show that giant nonequilibrium fluctuations are present during the free diffusive remixing occurring in ordinary liquid mixtures and in macromolecular solutions. The static structure factor of the fluctuations is measured by using a quantitative shadowgraph technique. We show that structure factors at different times and from different samples can be rescaled onto a single master curve without any adjustable parameter, thus giving strong evidence that nonequilibrium fluctuations are a universal feature of free diffusion processes. PMID- 11046359 TI - Multiscaling in inelastic collisions AB - We study relaxation properties of two-body collisions on the mean-field level. We show that this process exhibits multiscaling asymptotic behavior as the underlying distribution is characterized by an infinite set of nontrivial exponents. These nonequilibrium relaxation characteristics are found to be closely related to the steady state properties of the system. PMID- 11046360 TI - Dynamics of a penta-hepta defect in a hexagonal pattern AB - The structure and dynamics of a penta-hepta defect in a hexagonal pattern are studied experimentally. The hexagonal pattern is formed by placing a layer of soap bubbles (diameter approximately 1 mm) on a flat glass plate. We find that an isolated penta-hepta defect in a bubble raft with free boundary always moves along the direction perpendicular to the wave vector of the nonsingular mode and towards the nearest boundary. The structure of the penta-hepta defect is found to be similar to that found in nonequilibrium pattern forming systems. PMID- 11046361 TI - Large-q asymptotics of the random-bond potts model AB - We numerically examine the large-q asymptotics of the q-state random bond Potts model. Special attention is paid to the parametrization of the critical line, which is determined by combining the loop representation of the transfer matrix with Zamolodchikov's c-theorem. Asymptotically the central charge seems to behave like c(q)=1 / 2 log(2)(q)+O(1). Very accurate values of the bulk magnetic exponent x(1) are then extracted by performing Monte Carlo simulations directly at the critical point. As q-->infinity, these seem to tend to a nontrivial limit, x(1)-->0.192+/-0.002. PMID- 11046362 TI - Wave scattering through classically chaotic cavities in the presence of absorption: An information-theoretic model AB - We propose an information-theoretic model for the transport of waves through a chaotic cavity in the presence of absorption. The entropy of the S-matrix statistical distribution is maximized, with the constraint =alphan: n is the dimensionality of S, and 0>1 Rayleigh statistics is attained even with no absorption; here, we extend the study to alpha<1. The model is compared with random-matrix-theory numerical simulations: it describes the problem very well for strong absorption, but fails for moderate and weak absorptions. Thus, in the latter regime, some important physical constraint is missing in the construction of the model. PMID- 11046363 TI - Growth of taylor vortices: A molecular dynamics study AB - Molecular dynamics methods have been used in a quantitative study of the growth and decay of Taylor vortices in a fluid confined between concentric cylinders when the rotation of the inner cylinder is instantaneously started or stopped. Analysis of the temporal evolution of the vortex flow fields shows that the behavior of this microscopic system agrees with experiment. In order to make the analysis entirely self-contained, torque measurements have been used to determine the effective viscosity of the fluid. PMID- 11046364 TI - Abnormal rolls and regular arrays of disclinations in homeotropic electroconvection AB - We present the first quantitative verification of an amplitude description for systems with (nearly) spontaneously broken isotropy, in particular for the recently discovered abnormal-roll states. We also obtain a conclusive picture of the three-dimensional director configuration in a spatial period doubling phenomenon involving disclination loops. The first observation of two Lifshitz frequencies in electroconvection is reported. PMID- 11046365 TI - Inverse energy cascade in two-dimensional turbulence: deviations from gaussian behavior AB - High-resolution numerical simulations of stationary inverse energy cascade in two dimensional turbulence are presented. Deviations from Gaussian behavior of velocity differences statistics are quantitatively investigated. The level of statistical convergence is pushed enough to permit reliable measurement of the asymmetries in the probability distribution functions of longitudinal increments and odd-order moments, which bring the signature of the inverse energy flux. No measurable intermittency corrections could be found in their scaling laws. The seventh order skewness increases by almost two orders of magnitude with respect to the third, thus becoming of order unity. PMID- 11046366 TI - Intermittency of temperature field in turbulent convection AB - The scaling behavior of the temperature structure functions in turbulent convection is found to be different for length scales below and above the Bolgiano scale. Both sets of the exponents are well described by log-Poisson statistics. The parameter beta(T) which measures the degree of intermittency is the same for the two regimes of scales and is consistent with the corresponding value for the passive scalar field. A balance between thermal forcing and nonlinear velocity advection, which is a key ingredient leading to Bolgiano scaling, is also checked. PMID- 11046367 TI - Bilayer-by-bilayer antiferroelectric ordering in freely suspended films of an achiral polymer-monomer liquid crystal mixture AB - Thin freely suspended films of a mixture of an achiral side-chain liquid crystal polymer and its monomer have been studied with depolarized reflected light microscopy. We observe that regions with an odd number of bilayers exhibit a net spontaneous polarization in the tilt plane of the molecules, while regions with an even number of bilayers have no net polarization. These odd-even effects are direct evidence that the tilted smectic bilayers are anticlinic at the polymer backbone and synclinic at bilayer interface and confirm that the phase is bilayer by-bilayer antiferroelectric. PMID- 11046368 TI - Apparent finite-size effects in the dynamics of supercooled liquids AB - Molecular dynamics simulations are performed for a supercooled simple liquid with changing the system size from N=108 to 10(4) to examine possible finite-size effects. Although almost no systematic deviation is detected in the static pair correlation functions, it is demonstrated that the structural alpha relaxation in a small system becomes considerably slower than that in larger systems for temperatures below T(c) at which the size of the cooperative particle motions becomes comparable to the unit cell length of the small system. The discrepancy increases with decreasing temperature. PMID- 11046369 TI - Pumping liquids using asymmetric electrode arrays AB - Following a general symmetry argument, I suggest using locally asymmetric electric geometries to pump liquid in channels or drive droplets on surfaces. This strategy, which requires no global gradient in the pumping direction, should be of interest for microfluidic devices and micro-electro-mechanical systems. A practical realization consists in using polar periodic arrays of electrodes addressed by an ac voltage difference. A simple electro-osmotic model provides an estimate of the pumping velocities achievable. PMID- 11046370 TI - Velocity and shape selection of dendritic crystals in a forced flow AB - The phase-field method is used to simulate the two-dimensional growth of a dendritic crystal in a forced flow. The selection of the velocity and shape of the dendrite tip is investigated as a function of flow rate, growth direction relative to the flow, as well as anisotropy strength, and the results for the upstream growing tips are compared to existing theoretical predictions. PMID- 11046371 TI - Reductions of the glass transition temperature in thin polymer films: probing the length scale of cooperative dynamics AB - We report measurements of the glass transition temperature, T(g), in free standing polymer films in a low M(n) limit where chain confinement effects are not observed. The measured T(g) values are accurately described by a layer model incorporating a mobile surface layer with a size determined by the length scale of cooperative dynamics. The analysis leads to a surface T(g) value and length scale of cooperative motion near bulk T(g) which quantitatively agree with independently determined values. The model and parameters provide a framework within which all previous measurements of T(g) values in thin supported films may be understood and provides values for the length scale of cooperative motion over an extended range of temperatures below the bulk T(g) value. PMID- 11046372 TI - Phase separation and shape deformation of two-phase membranes AB - Within a coupled-field Ginzburg-Landau model we study analytically phase separation and accompanying shape deformation on a two-phase elastic membrane in simple geometries such as cylinders, spheres, and tori. Using an exact periodic domain wall solution we solve for the shape and phase separating field, and estimate the degree of deformation of the membrane. The results are pertinent to preferential phase separation in regions of differing curvature on a variety of vesicles. PMID- 11046374 TI - Continuum approach to car-following models AB - A continuum version of the car-following Bando model is developed using a series expansion of the headway in terms of the density. This continuum model obeys the same stability criterion as its discrete counterpart. To compare both models we show that traveling wave solutions of the Bando model are very similar to those of the continuum model in the limit of small changes of headway. As the change of headway across the wave increases the solutions gradually diverge. Our transformation relating headway to density enables predictions of the global impact and characteristics of any car-following model using the analogous continuum model. In contrast, we show that the conventional continuum models which account for effects of pressure and dispersion predict behavior which is distinct from the global behavior of discrete models. PMID- 11046373 TI - Continuous probability distributions from finite data. AB - Recent approaches to the problem of inferring a continuous probability distribution from a finite set of data have used a scalar field theory for the form of the prior probability distribution. This paper presents a more general form for the prior distribution that has a geometrical interpretation and can improve the specificity of likely solutions. It is also demonstrated that a numerical sampling of the posterior probability distribution can be used as an alternative to a histogram for visualization and to make probabilistic inferences from the data. PMID- 11046375 TI - Self-organized criticality in a computer network model AB - We study the collective behavior of computer network nodes by using a cellular automaton model. The results show that when the load of network is constant, the throughputs and buffer contents of nodes are power-law distributed in both space and time. Also the feature of 1/f noise appears in the power spectrum of the change of the number of nodes that bear a fixed part of the system load. It can be seen as yet another example of self-organized criticality. Power-law decay in the distribution of buffer contents implies that heavy network congestion occurs with small probability. The temporal power-law distribution for throughput might be a reasonable explanation for the observed self-similarity in computer network traffic. PMID- 11046376 TI - Elastic constants from microscopic strain fluctuations AB - Fluctuations of the instantaneous local Lagrangian strain epsilon(ij)(r,t), measured with respect to a static "reference" lattice, are used to obtain accurate estimates of the elastic constants of model solids from atomistic computer simulations. The measured strains are systematically coarse-grained by averaging them within subsystems (of size L(b)) of a system (of total size L) in the canonical ensemble. Using a simple finite size scaling theory we predict the behavior of the fluctuations as a function of L(b)/L and extract elastic constants of the system in the thermodynamic limit at nonzero temperature. Our method is simple to implement, efficient, and general enough to be able to handle a wide class of model systems, including those with singular potentials without any essential modification. We illustrate the technique by computing isothermal elastic constants of "hard" and "soft" disk triangular solids in two dimensions from Monte Carlo and molecular dynamics simulations. We compare our results with those from earlier simulations and theory. PMID- 11046377 TI - Invariant power law distribution of langevin systems with colored multiplicative noise AB - The random multiplicative process is studied for the case of a colored multiplicative noise with exponentially decreasing autocorrelation function. We observe the power law exponent of probability distribution in a statistically steady state numerically to clarify the effect of finite correlation time. The renormalization procedure is applied to derive the power law exponent theoretically. The power law exponent is inversely proportional to the autocorrelation time of the multiplicative noise. PMID- 11046378 TI - Perturbation expansion in phase-ordering kinetics. II. N-vector model AB - The perturbation theory expansion presented earlier to describe the phase ordering kinetics in the case of a nonconserved scalar order parameter is generalized to the case of the n-vector model. At lowest order in this expansion, as in the scalar case, one obtains the theory due to Ohta, Jasnow, and Kawasaki (OJK). The second-order corrections for the nonequilibrium exponents are worked out explicitly in d dimensions and as a function of the number of components n of the order parameter. In the formulation developed here the corrections to the OJK results are found to go to zero in the large n and d limits. Indeed, the large-d convergence is exponential. PMID- 11046379 TI - Delocalization transition of a rough adsorption-reaction interface AB - We introduce a kinetic interface model suitable for simulating adsorption reaction processes which take place preferentially at surface defects such as steps and vacancies. As the average interface velocity is taken to zero, the self affine interface with Kardar-Parisi-Zhang-like scaling behavior undergoes a delocalization transition with critical exponents that fall into a different universality class. As the critical point is approached, the interface becomes a multivalued, multiply connected self-similar fractal set. The scaling behavior and critical exponents of the relevant correlation functions are determined from Monte Carlo simulations and scaling arguments. PMID- 11046380 TI - Localized structures in nonlinear lattices with diffusive coupling and external driving AB - We study the stabilization of localized structures by discreteness in one dimensional lattices of diffusively coupled nonlinear sites. We find that in an external driving field these structures may lose their stability by either relaxing to a homogeneous state or nucleating a pair of oppositely moving fronts. The corresponding bifurcation diagram demonstrates a cusp singularity. The obtained analytic results are in good quantitative agreement with numerical simulations. PMID- 11046381 TI - Diffusion-mediated reactions with a time-dependent absorption rate AB - Diffusion-mediated reactions models are particularly useful for the characterization of physical, chemical, and biological problems. In this paper we present a theoretical study of the absorption probability density, survival probability, and reaction rate for diffusion-mediated reactions models with a time-dependent finite absorption rate (an extension of a model usually referred to as the "imperfect trap model"). The results are obtained by means of the formalism of continuous time random walk on a lattice and considering a general reaction dynamics upon encounter of the reactives. First jump probability densities are included to take initial conditions into account. Previous results presented by Collins and Kimball [J. Colloid. Sci. 4, 425 (1949)] and Noyes [J. Chem. Phys. 22, 1349 (1954)] are reobtained for the particular case of a time independent absorptivity. Short and long time behaviors are analyzed resulting, in particular, in that the long time behavior of the absorption probability density exhibits the same time dependence as the first passage time density. The results obtained are illustrated by considering a one-dimensional model with consequent discussion. PMID- 11046382 TI - Ultrametricity in three-dimensional edwards-anderson spin glasses AB - We perform an accurate test of ultrametricity in the aging dynamics of the three dimensional Edwards-Anderson spin glass. Our method consists in considering the evolution in parallel of two identical systems constrained to have fixed overlap. This turns out to be a particularly efficient way to study the geometrical relations between configurations at distant large times. Our findings strongly hint towards dynamical ultrametricity in spin glasses, while this is absent in simpler aging systems with domain growth dynamics. A recently developed theory of linear response in glassy systems allows us to infer that dynamical ultrametricity implies the same property at the level of equilibrium states. PMID- 11046383 TI - Diffusion over a saddle with a langevin equation AB - The diffusion problem over a saddle is studied using a multidimensional Langevin equation. An analytical solution is derived for a quadratic potential and the probability to pass over the barrier deduced. A very simple solution is given for the one-dimensional problem and a general scheme is shown for higher dimensions. PMID- 11046384 TI - Competitive reactions among three monomers over a catalytic surface AB - We studied in this work a three-monomer reaction model on one- and two dimensional lattices. We have taken different reactivity rates among pairs of monomers and the reaction between two selected monomers was forbidden. We have employed the mean field and the pair approximation to decouple the equations of motion for the densities of single and pairs of monomers. We found the stationary states and the phase diagram of the model. We have shown that, in two dimensions and within the pair approximation, there is a first-order transition line between active and poisoned steady states. PMID- 11046385 TI - Inequivalence of dynamical ensembles in a generalized driven diffusive lattice gas AB - We generalize the driven diffusive lattice gas model by using a combination of Kawasaki and Glauber dynamics. We find via Monte Carlo simulations and perturbation studies that the simplest possible generalization of the equivalence of the canonical and grand-canonical ensembles, which holds in equilibrium, does not apply for this class of nonequilibrium systems. PMID- 11046386 TI - Equilibrium and stationary nonequilibrium states in a chain of colliding harmonic oscillators AB - Equilibrium and nonequilibrium properties of a chain of colliding harmonic oscillators (ding-dong model) are investigated. Our chain is modeled as harmonically bounded particles that can only interact with neighboring particles by hard-core interaction. Between the collisions, particles are just independent harmonic oscillators. We are especially interested in the stationary nonequilibrium state of the ding-dong model coupled with two stochastic heat reservoirs (not thermostated) at the ends, whose temperature is different. We check the Gallavotti-Cohen fluctuation theorem [G. Gallavoti and E. G. D. Cohen, Phys. Rev. Lett. 74, 2694 (1995)] and also the Evans-Searles identity [D. Evans and D. Searles, Phys. Rev. E. 50, 1994 (1994)] numerically. It is verified that the former theorem is satisfied for this system, although the system is not a thermostated system. PMID- 11046387 TI - Background estimation in experimental spectra AB - A general probabilistic technique for estimating background contributions to measured spectra is presented. A Bayesian model is used to capture the defining characteristics of the problem, namely, that the background is smoother than the signal. The signal is allowed to have positive and/or negative components. The background is represented in terms of a cubic spline basis. A variable degree of smoothness of the background is attained by allowing the number of knots and the knot positions to be adaptively chosen on the basis of the data. The fully Bayesian approach taken provides a natural way to handle knot adaptivity and allows uncertainties in the background to be estimated. Our technique is demonstrated on a particle induced x-ray emission spectrum from a geological sample and an Auger spectrum from iron, which contains signals with both positive and negative components. PMID- 11046388 TI - Renormalized field theory of driven lattice gases under infinitely fast drive AB - We use field theoretic renormalization group methods to study the critical behavior of a recently proposed Langevin equation for driven lattice gases under infinitely fast drive. We perform an expansion around the upper critical dimension, d(c)=4, and obtain the critical exponents to one-loop order. The main features of the two-loop calculation are also outlined. The renormalized theory is shown to exhibit a behavior different from the standard field theory for the driven lattice gas with finite driving, i.e., it is not mean-field-like. PMID- 11046389 TI - One-dimensional stochastic Levy-lorentz gas AB - We introduce a Levy-Lorentz gas in which a light particle is scattered by static point scatterers arranged on a line. We investigate the case where the intervals between scatterers xi(i) are independent random variables identically distributed according to the probability density function &mgr;(xi) approximately xi( (1+gamma)). We show that under certain conditions the mean square displacement of the particle obeys >/=Ct3-gamma for 1/=1) representing the degree of breaking of the conservation law. This model is reduced to be the BTW model when alpha=1. By calculating the average number of topplings in an avalanche exactly, it is shown that for any alpha>1, with an exponent 1 as alpha-->1 gives a scaling relation 2nu(2-a)=1 for the critical exponents nu and a of the distribution function of T. The 1-1 height correlation C11(r) is also calculated analytically and we show that C11(r) is bounded by an exponential function when alpha>1, although C11(r) approximately r(-2d) was proved by Majumdar and Dhar for the d dimensional BTW model. A critical exponent nu(11) characterizing the divergence of the correlation length xi as alpha-->1 is defined as xi approximately |alpha 1|(-nu(11)) and our result gives an upper bound nu(11)]. PMID- 11046404 TI - Static A+B-->0 annihilation process in arbitrary dimension AB - The pair annihilation A+B-->0 of static particles distributed at random in a d dimensional space is studied in the large time regime for a tunneling law. The consequences of a superposition approximation used to close the hierarchy of equations describing the process are analytically explored. It is shown that the density and pair correlations of surviving particles have scaling expressions which display the ordering and clustering effects. PMID- 11046405 TI - Three-dimensional effects in directional solidification in hele-shaw cells: nonlinear evolution and pattern selection AB - Directional solidification of a dilute binary alloy in a Hele-Shaw cell is modeled by a long-wave nonlinear evolution equation with zero flux and contact angle conditions at the walls. The basic steady-state solution and its linear stability criteria are found analytically, and the nonlinear system is solved numerically. Concave-down (toward the solid) interfaces under physically realistic conditions are found to be more unstable than the planar front. Weakly nonlinear analysis indicates that subcritical bifurcation is promoted, the domain of modulational instability is expanded and transition to three-dimensional patterns is delayed due to the contact-angle condition. In the strongly nonlinear regime fully three-dimensional steady-state solutions are found whose characteristic amplitude is larger than that for the two-dimensional problem. In the subcritical regime secondary bifurcation to stable solutions is promoted. PMID- 11046406 TI - Diagrammatic approach for open chaotic systems AB - A semiclassical diagrammatic approach is constructed for calculating correlation functions of observables in open chaotic systems with time reversal symmetry. The results are expressed in terms of classical correlation functions involving Wigner representations of the observables. The formalism is used to explain a recent microwave experiment on the four-disk problem, and to characterize the two point function of the photodissociation cross section of complex molecules. PMID- 11046407 TI - Numerical investigation of the effects of classical phase space structure on a quantum system with decoherence AB - We present a detailed numerical study of a chaotic classical system and its quantum counterpart. The system is a special case of a kicked rotor and for certain parameter values possesses cantori dividing chaotic regions of the classical phase space. We investigate the diffusion of particles through a cantorus. A quantum analysis confirms that the cantori act as barriers. We numerically estimate the classical phase space flux through the cantorus per kick and relate this quantity to the behavior of the quantum system. We introduce decoherence via environmental interactions with the quantum system and observe the subsequent increase in the transport of quantum particles through the boundary. PMID- 11046408 TI - Field-induced transitions and spatial chaos in the classical XY spin chain AB - We study the lowest-lying excitation of a classical ferromagnetic XY spin chain, in the presence of a symmetry breaking magnetic field. Extremizing the energy of this system leads to a two-dimensional nonlinear map, whose allowed phase space shrinks with increasing field in a nontrivial manner. The orbits of the map represent the set of extremum energy spin configurations. For each field, we compute the energy of the members of this set and find the lowest energy among them, excluding the obvious ground state configuration with all spins parallel along the field direction. This state turns out to be the unstable fixed point of the map. We show that up to a certain (primary) critical field, a separatrixlike 2pi soliton configuration is the lowest-energy excitation, with an energy very close to the ground state energy. For any field beyond this critical field, the soliton disappears and lowest excitation is a librational configuration corresponding to the outermost orbit in the phase plot at that field. Further, its energy is found to be much higher than the ground state energy, leading to a sharp jump in the difference in energy between the former and the latter at this field. With further increase in the field, sharp jumps in the excitation energy arise at certain secondary critical fields as well. We show that these appear when the corresponding spin configurations become commensurate. This complex behavior of the energy is interpreted and its effect on the magnetization and static susceptibility of the system is also studied. PMID- 11046409 TI - Strange attractor for the renormalization flow for invariant tori of hamiltonian systems with two generic frequencies AB - We analyze the stability of invariant tori for Hamiltonian systems with two degrees of freedom by constructing a transformation that combines Kolmogorov Arnold-Moser theory and renormalization-group techniques. This transformation is based on the continued fraction expansion of the frequency of the torus. We apply this transformation numerically for arbitrary frequencies that contain bounded entries in the continued fraction expansion. We give a global picture of renormalization flow for the stability of invariant tori, and we show that the properties of critical (and near critical) tori can be obtained by analyzing renormalization dynamics around a single hyperbolic strange attractor. We compute the fractal diagram, i.e., the critical coupling as a function of the frequencies, associated with a given one-parameter family. PMID- 11046410 TI - One-dimensional dynamics for traveling fronts in coupled map lattices AB - Multistable coupled map lattices typically support traveling fronts, separating two adjacent stable phases. We show how the existence of an invariant function describing the front profile allows a reduction of the infinitely dimensional dynamics to a one-dimensional circle homeomorphism, whose rotation number gives the propagation velocity. The mode locking of the velocity with respect to the system parameters then typically follows. We study the behavior of fronts near the boundary of parametric stability, and we explain how the mode locking tends to disappear as we approach the continuum limit of an infinite density of sites. PMID- 11046411 TI - Lyapunov exponents and kolmogorov-sinai entropy for a high-dimensional convex billiard AB - We compute the Lyapunov exponents and the Kolmogorov-Sinai (KS) entropy for a self-bound N-body system that is realized as a convex billiard. This system exhibits truly high-dimensional chaos, and 2N-4 Lyapunov exponents are found to be positive. The KS entropy increases linearly with the numbers of particles. We examine the chaos generating defocusing mechanism and investigate how high dimensional chaos develops in this system with no dispersing elements. PMID- 11046412 TI - Linear and nonlinear time series analysis of the black hole candidate cygnus X-1 AB - We analyze the variability in the x-ray lightcurves of the black hole candidate Cygnus X-1 by linear and nonlinear time series analysis methods. While a linear model describes the overall second order properties of the observed data well, surrogate data analysis reveals a significant deviation from linearity. We discuss the relation between shot noise models usually applied to analyze these data and linear stochastic autoregressive models. We debate statistical and interpretational issues of surrogate data testing for the present context. Finally, we suggest a combination of tools from linear and nonlinear time series analysis methods as a procedure to test the predictions of astrophysical models on observed data. PMID- 11046413 TI - Estimating generating partitions of chaotic systems by unstable periodic orbits AB - An outstanding problem in chaotic dynamics is to specify generating partitions for symbolic dynamics in dimensions larger than 1. It has been known that the infinite number of unstable periodic orbits embedded in the chaotic invariant set provides sufficient information for estimating the generating partition. Here we present a general, dimension-independent, and efficient approach for this task based on optimizing a set of proximity functions defined with respect to periodic orbits. Our algorithm allows us to obtain the approximate location of the generating partition for the Ikeda-Hammel-Jones-Moloney map. PMID- 11046414 TI - Control of transient chaos in tent maps near crisis. I. Fixed point targeting AB - Combinatorial techniques are applied to the symbolic dynamics representing transient chaotic behavior in tent maps in order to solve the problem of Ott Grebogi-Yorke control to the nontrivial fixed point occurring in such maps. This approach allows "preimage overlap" to be treated exactly. Closed forms for both the probability of control being achieved and the average number of iterations to control are derived. The results are discussed in relation to the work of Tel and shed new light on the transition to the control of permanent chaos. PMID- 11046415 TI - Control of transient chaos in tent maps near crisis. II. Periodic orbit targeting AB - Recent work on a symbolic approach to the calculation of probability distributions arising in the application of the Ott-Grebogi-Yorke strategy to transiently chaotic tent maps is extended to the case of control to a nontrivial periodic orbit. Closed forms are derived for the probability of control being achieved and the average number of iterations to control when it occurs. Both single-component and multiple-component targeting are considered, and illustrative examples of the results obtained are presented. PMID- 11046416 TI - Karhunen-Loeve local characterization of spatiotemporal chaos in a reaction diffusion system AB - By computing the Karhunen-Loeve decomposition (KLD) correlation length xi(KLD) of a reaction-diffusion system in the extensive chaos regime, we show that it is a sensitive measure of spatial dynamical inhomogeneities. It reveals substantial spatial nonuniformity of the dynamics at the boundaries and can also detect slow spatial variations in system parameters. The intensive length xi(KLD) can be easily computed from small local subsystems and is found to have a similar parametric dependence as the two-point correlation length computed over the full system size. PMID- 11046417 TI - Intermittency and dynamical chaos in reversible spontaneous emission AB - It is shown that a type of reversible spontaneous emission, the chaotic vacuum Rabi oscillations, may occur in the interaction of two-level atoms strongly coupled with a single cavity mode under a modulation of the atom-field coupling. Such a modulation arises naturally if the atoms move through a cavity in maserlike experiments. The existence of homoclinic chaos in reversible spontaneous emission is proven analytically. Evidence of intermittency associated with the spatial modulation of the vacuum Rabi frequency is shown numerically for the values of parameters that are achievable in present-day experiments with Rydberg atoms moving through a high-Q microwave cavity in the strong-coupling regime. PMID- 11046418 TI - Extended local balance model of turbulence and couette-taylor flow AB - An extended local balance model of turbulence, based on a new transport equation for the dissipation rate with a negative diffusion coefficient, is presented. Analytical solutions for the mean velocity and the dissipation rate for the turbulent Couette-Taylor problem are derived. The dependence of torque on the Reynolds number is obtained. These solutions depend only on two constants k=0.4 and C=9.5 of the turbulent boundary layer and, within the limits of a narrow channel, are reduced to the well-known von Karman's solutions for planar Couette flow. Strange attractor behavior in this limit is also observed. PMID- 11046419 TI - Probability distribution functions of derivatives and increments for decaying burgers turbulence AB - A Lagrangian method is used to show that the power law with a -7/2 exponent in the negative tail of the probability distribution function (PDF) of the velocity gradient and of velocity increments, predicted by E et al. [Phys. Rev. Lett. 78, 1904 (1997)] for forced Burgers turbulence, is also present in the unforced case. The theory is extended to the second-order space derivative whose PDF has power law tails with exponent -2 at both large positive and negative values and to the time derivatives. PDF's of space and time derivatives have the same (asymptotic) functional forms. This is interpreted in terms of a random Taylor hypothesis. PMID- 11046420 TI - Logarithmically slow expansion of hot bubbles in gases AB - We predict a logarithmically slow expansion of hot bubbles in gases in the process of cooling. A model problem is first solved, when the temperature has compact support. Then the temperature profile decaying exponentially at large distances is considered. The periphery of the bubble is shown to remain essentially static ("glassy") in the process of cooling until it is taken over by a logarithmically slowly expanding "core." An analytical solution to the problem is obtained by matched asymptotic expansion. This problem gives an example of how logarithmic corrections enter dynamic scaling. PMID- 11046421 TI - Decaying magnetohydrodynamics: effects of initial conditions AB - We study the effects of homogenous and isotropic initial conditions on decaying magnetohydrodynamics (MHD). We show that for an initial distribution of velocity and magnetic-field fluctuations, appropriately defined structure functions decay as a power law in time. We also show that for a suitable choice of initial cross correlations between velocity and magnetic fields even-order structure functions acquire anomalous scaling in time where as scaling exponents of the odd-order structure functions remain unchanged. We discuss our results in the context of fully developed MHD turbulence. PMID- 11046422 TI - Anomalous diffusion with absorption: exact time-dependent solutions AB - Recently, analytical solutions of a nonlinear Fokker-Planck equation describing anomalous diffusion with an external linear force were found using a nonextensive thermostatistical Ansatz. We have extended these solutions to the case when an homogeneous absorption process is also present. Some peculiar aspects of the interrelation between the deterministic force, the nonlinear diffusion, and the absorption process are discussed. PMID- 11046423 TI - Spinodal decomposition of two-dimensional fluid mixtures: A spectral analysis of droplet growth AB - The spinodal decomposition of two-dimensional fluid mixture is studied by numerical simulation. For the high viscous fluid mixture it has not been evident whether the interfacial tension is relevant to the droplet growth or not. A length scale R defined by the structure function extracting the effect of the long wavelength mode justifies a rapid growth close to R approximately t, but the length scale energetically defined reveals a much slower growth R approximately t(0.5), where t is time. This discrepancy represents the violation of the dynamical scaling with single length scale. The slow gowth of the length scale is attributed to the accumulation of the number of isolated droplets in phase separating state, whereas the rapid growth represents the relevance of the surface tension as the driving force in two dimensions. For a low viscous fluid mixture the dynamical scaling is a good assumption with the growth law R approximately t(2/3) up to a very large Reynolds number Re approximately 1500, which is the limit in the present simulation. PMID- 11046424 TI - Structure and dynamics of nanofluids: theory and simulations to calculate viscosity AB - The simplified expression of the Pozhar-Gubbins (PG) rigorous, nonequilibrium statistical mechanical theory of dense, strongly inhomogeneous fluids is used to calculate the viscosity of model fluids confined in a slit pore of several molecular diameters in width in terms of the equilibrium structure factors (i.e., the number density and pair correlation functions) of these nanofluids obtained by means of the equilibrium molecular dynamic simulations. These results are compared to those obtained by means of the nonequilibrium molecular dynamic simulations of the planar Poiseuille flow of the model nanofluids, and to the results supplied by several heuristic expressions for the nanofluid viscosity. This comparison proves that the PG transport theory provides a reliable, quantitatively accurate description of the viscosity coefficients of the model nanofluids while all the heauristic approaches fail. This success of the PG prediction of the nanofluid viscosity is because the theoretical expression accounts accurately for the nanofluid structure. PMID- 11046425 TI - Small scale intermittency and bursting in a turbulent channel flow AB - The statistical properties of the streamwise velocity fluctuations in a fully developed turbulent channel flow are studied experimentally by means of single hot wire measurements. The intermittency features, studied through the scaling of the moments of the velocity structure function computed using the extended self- similarity and through the probability density function of the wavelet coefficients, are found to be dependent on the distance from the wall. The maximum intermittency effects are observed in the region between the buffer layer and the inner part of the logarithmic region where it is known that the bursting phenomenon, related to coherent structures such as low speed streaks and streamwise vortices, is the dominant dynamical feature. An eduction technique based on wavelet transform for identification of organized motion is developed and used to analyze the turbulent signals. Streamwise velocity conditional averages computed on events educed with the proposed method are reported. Events responsible for intermittency are found to consist of regions of high velocity gradients and are directly correlated with the observed increase of intermittency close to the wall. PMID- 11046426 TI - Shear-thinning-induced chaos in taylor-couette flow AB - The effect of weak shear thinning on the stability of the Taylor-Couette flow is explored for a Carreau-Bird fluid in the narrow-gap limit. The Galerkin projection method is used to derive a low-order dynamical system from the conservation of mass and momentum equations. In comparison with the Newtonian system, the present equations include additional nonlinear coupling in the velocity components through the viscosity. It is found that the critical Taylor number, corresponding to the loss of stability of the base (Couette) flow, becomes lower as the shear-thinning effect increases. That is, shear thinning tends to precipitate the onset of Taylor vortex flow. Similar to Newtonian fluids, there is an exchange of stability between the Couette and Taylor vortex flows, which coincides with the onset of a supercritical bifurcation. However, unlike the Newtonian model, the Taylor vortex cellular structure loses its stability in turn as the Taylor number reaches a critical value. At this point, a Hopf bifurcation emerges, which exists only for shear-thinning fluids. PMID- 11046427 TI - Trapped rossby waves AB - The possibility of tidal dynamics at strictly imaginary Lamb parameters has been known for more than three decades. The present paper explores the prevailing physics in this parameter regime. To this end, basic features of the global circulation such as baroclinicity and geostrophy have to be incorporated into tidal dynamics. The tidal equations of the thermal wind are readily obtained in the framework of spherical bishallow water theory. Density surfaces of a circulation with available potential energy alter the spatial inhomogenities of the generic tidal problem. Wave dynamics in an inhomogeneous medium are characterized not only by a dispersion relation but also by a wave guide geography: significant wave amplitudes are trapped in specific regions of frequency-dependent width. As an inherently global issue, evaluation of the Rossby wave guide geography for a given circulation cannot rely on the familiar regional filters of tidal theory. On the global domain, the Rossby wave specification is given by the Margules filter. A thermal wind is stable against nondivergent Rossby wave disturbances. Rossby waves propagating with a geostrophic wind are governed by prolate dynamics (real Lamb parameters) while imaginary Lamb parameters emerge for the oblate dynamics of Rossby waves running against a geostrophic wind. Oblate Rossby wave dynamics include pole-centered wave guides and very low-frequency disturbances propagating eastward against a westward wind. PMID- 11046428 TI - Effect of slow compression on the linear stability of an accelerated shear layer AB - An analysis is given of the effect of a slow uniform anisotropic compression or expansion on the linear stability of a normally accelerated planar interface between two fluids with different densities and tangential velocities, i.e., a combined Kelvin-Helmholtz and Rayleigh-Taylor instability, but generalized to an arbitrary time-dependent acceleration history. The compression is presumed to be sufficiently slow that the density remains uniform within each fluid and hence depends only on time. The perturbation is taken to be sinusoidal with amplitude h(t). The time evolution of h is determined by requiring pressure continuity across the interface in the usual way. The resulting linearized stability equation is a second-order linear ordinary differential equation for h(t). Compared to the corresponding well-known result for incompressible fluids, it is found that normal compression has the effect of reducing the perturbation growth rate &hdot; by an obvious geometrical correction, while transverse compression does not directly affect the net growth rate but rather has the dynamical effect of increasing its time derivative. When attention is focused on the masses transported across the initial interface rather than h, the purely geometrical effects of compression no longer appear explicitly, while the dynamical effects remain. It is thereby shown that both normal and transverse compression dynamically enhance the mixing of material masses, in spite of the corresponding purely geometrical reduction in &hdot;. PMID- 11046429 TI - Ultrasound scattering and the study of vortex correlations in disordered flows AB - In an idealized way, some turbulent flows can be pictured by assemblies of many vortices characterized by a set of particle distribution functions. Ultrasound provides a useful, nonintrusive, tool to study the spatial structure of vorticity in flows. This is analogous to the use of elastic neutron scattering to determine liquid structure. We express the dispersion relation, as well as the scattering cross section, of sound waves propagating in a "liquid" of identical vortices as a function of vortex pair correlation functions. In two dimensions, formal analogies with ionic liquids are pointed out. PMID- 11046430 TI - Luminescence from acoustic-driven laser-induced cavitation bubbles AB - The influence of a continuous sound field on the first oscillation cycle and on the cavitation luminescence of a transient laser-induced bubble is investigated experimentally. The variation of the collapse phase is predicted with a simple numerical model and compared with experiment. Bubble dynamics is mainly influenced by three parameters: the phase of bubble generation, the size of the bubble, and the amplitude of the sound field. The experimentally found enhancement and reduction of the luminescence is discussed and several suggestions are made for further boosting of the collapse strength. PMID- 11046431 TI - Cavity formation and the drying transition in the lennard-jones fluid AB - By simulation and theory, we study the probability of observing N molecular centers within molecular sized volumes for a Lennard-Jones fluid near liquid vapor coexistence. For large volumes and small N, the probability distribution differs markedly from Gaussian. The free energy per unit surface area to form empty volumes (i.e., cavities) is a rapidly varying function of the radius for small cavities. It becomes constant for large volumes. The source of these behaviors is the occurrence of drying (i.e., solvent depletion) at the cavity surface. The crossover to drying occurs on microscopic length scales, with significant density depletion found for cavities with radii of the order of two or more Lennard-Jones diameters. Reasonable agreement is found between the simulation results and the theory developed by Lum, Chandler, and Weeks [J. Phys. Chem. B 103, 4570 (1999)]. PMID- 11046432 TI - Thermal and inertial modes of convection in a rapidly rotating annulus AB - The nature of the primary instabilities that arise in a fluid contained in a fast rotating cylindrical annulus with slightly inclined plane top and bottom boundaries, radial gravity, and internal heating is numerically analyzed. It is shown that for moderate and high Prandtl numbers, the onset of convection is described by a competition of azimuthal thermal modes with different radial structure, which dominate in different regions of the parameter space. By the combined effect of the inclined ends and rotation, there are modes that are attached to the heated wall and slanted to the prograde direction of rotation, and others which are straight and fill the convective layer. Nevertheless, for very small Prandtl numbers the velocity field of the dominant modes corresponds essentially to the inertial solution of the Poincare equation, and the temperature perturbation is forced by this velocity field. In addition, a detailed exploration of the critical Rayleigh numbers and precession frequencies of the convective modes versus the radius ratio and the Coriolis parameter, for different Prandtl numbers, is presented. PMID- 11046433 TI - Analysis of inelastic x-ray scattering spectra of low-temperature water AB - We analyze a set of high-resolution inelastic x-ray scattering (IXS) spectra from H2O measured at T=259, 273, and 294 K using two different phenomenological models. Model I, called the "dynamic cage model," combines the short time in-cage dynamics described by a generalized Enskog kinetic theory with a long-time cage relaxation dynamics described by an alpha relaxation. This model is appropriate for supercooled water where the cage effect is dominant and the existence of an alpha relaxation is evident from molecular-dynamics (MD) simulation data of extended simple point charge (SPC/E) model water. Model II is essentially a generalized hydrodynamic theory called the "three effective eigenmode theory" by de Schepper et al. 11. This model is appropriate for normal liquid water where the cage effect is less prominent and there is no evidence of the alpha relaxation from the MD data. We use the model I to analyze IXS data at T=259 K (supercooled water). We successfully extract the Debye-Waller factor, the cage relaxation time from the long-time dynamics, and the dispersion relation of high frequency sound from the short time dynamics. We then use the model II to analyze IXS data at all three temperatures, from which we are able to extract the relaxation rate of the central mode and the damping of the sound mode as well as the dispersion relation for the high-frequency sound. It turns out that the dispersion relations extracted from the two models at their respective temperatures agree with each other giving the high-frequency sound speed of 2900+/-300 m/s. This is to be compared with a slightly higher value reported previously, 3200+/-320 m/s, by analyzing similar IXS data with a phenomenological damped harmonic oscillator model 22. This latter model has traditionally been used exclusively for the analysis of inelastic scattering spectra of water. The k dependent sound damping and central mode relaxation rate extracted from our model analyses are compared with the known values in the hydrodynamic limit. PMID- 11046434 TI - Self-organization in nonlinear wave turbulence AB - We present a statistical equilibrium model of self-organization in a class of focusing, nonintegrable nonlinear Schrodinger (NLS) equations. The theory predicts that the asymptotic-time behavior of the NLS system is characterized by the formation and persistence of a large-scale coherent solitary wave, which minimizes the Hamiltonian given the conserved particle number (L2-norm squared), coupled with small-scale random fluctuations, or radiation. The fluctuations account for the difference between the conserved value of the Hamiltonian and the Hamiltonian of the coherent state. The predictions of the statistical theory are tested against the results of direct numerical simulations of NLS, and excellent qualitative and quantitative agreement is demonstrated. In addition, a careful inspection of the numerical simulations reveals interesting features of the transitory dynamics leading up to the long-time statistical equilibrium state starting from a given initial condition. As time increases, the system investigates smaller and smaller scales, and it appears that at a given intermediate time after the coalescense of the soliton structures has ended, the system is nearly in statistical equilibrium over the modes that it has investigated up to that time. PMID- 11046435 TI - Viscoelastic theory for nematic interfaces AB - A complete macroscopic theory for compressible nematic-viscous fluid interfaces is developed and used to characterize the interfacial elastic, viscous, and viscoelastic material properties. The derived expression for the interfacial stress tensor includes elastic and viscous components. Surface gradients of the interfacial elastic stress tensor generates tangential Marangoni forces as well as normal forces. The latter may be present even in planar surfaces, implying that in principle static planar interfaces may accommodate pressure jumps. The asymmetric interfacial viscous stress tensor takes into account the surface nematic ordering and is given in terms of the interfacial rate of deformation and interfacial Jaumann derivative. The material function that describes the anisotropic viscoelasticity is the dynamic interfacial tension, which includes the interfacial tension and dilational viscosities. Viscous dissipation due to interfacial compressibility is described by the anisotropic dilational viscosity, and it is shown to describe the Boussinesq surface fluid appropriate for Newtonian interfaces when the director is homeotropic. Three characteristic interfacial shear viscosities are defined according to whether the surface orientation is along the velocity direction, the velocity gradient, or the unit normal. In the last case the expression reduces to the interfacial shear viscosity of the Boussinesq surface fluid. The theory provides a theoretical framework to study interfacial stability, thin liquid film stability and hydrodynamics, and any other interfacial rheology phenomena. PMID- 11046436 TI - Local measurement of the zenithal anchoring strength AB - We present an electro-optic method for measuring the zenithal anchoring strength of nematic liquid crystals, based on the determination of the distortion produced by a small electric field. This method yields the zenithal anchoring strength at small applied torques, and remarkably, only needs local measurements (optical path difference versus applied voltage, sample thickness), in contrast to the classical methods that use measurements integrated over the entire sample. We determine the zenithal anchoring strength for two nematic liquid crystals (5 CB and 5 OCB) with positive dielectric anisotropy, onto poly(tetrafluoroethylene) (PTFE) treated surfaces, that yield planar liquid crystal cells. We find that the anchoring at the PTFE-5 CB interface is strong, with an extrapolation length approximately 30 nm, and independent of temperature far enough from the isotropic transition. We observe a pretransitional weakening of the anchoring strength near the nematic-isotropic transition, due to the reduction of the orientational order parameter at the interface. With 5 OCB, we measure a stronger anchoring, with an extrapolation length approximately 15 nm. This result may be explained by the increase of the van der Waals interactions between the liquid crystal molecules and the surface, due to the presence of the oxygen atom. PMID- 11046437 TI - Conformational dynamics of a metallomesogen studied by 2H-NMR spectroscopy AB - In this work we present a quantitative analysis of both quadrupolar splittings and deuterium Zeeman and quadrupolar spin-lattice relaxation times reported in the literature for two isotopomers of Azpac, an acetylacetonate derivative of the cyclopalladated 4, 4'-bis(hexyloxy) azoxybenzene. Azpac-d(4) is deuterated at the aromatic rings and Azpac-d(26) is deuterated on the alkoxy chains. The additive potential method is used to model the splittings, while the derived spectral densities are interpreted using the decoupled model in conjunction with the Nordio model. The two side chains are assumed to be noninteracting and identical in their conformations in order to limit the size of the transition rate matrix needed to describe correlated internal bond rotations in the chains. Rotational diffusion constants and internal jump rate constants are derived for this metallomesogen. PMID- 11046438 TI - Freedericksz transition in polymer-stabilized nematic liquid crystals AB - We have constructed polymer-stabilized nematic liquid crystals by photopolymerizing diacrylate monomers in the nematic phase. The orientation of the liquid crystal was controlled by the polymer network. We studied the Freedericksz transition in these systems. Experimentally we studied the transition by measuring the capacitance of the liquid crystal cells as a function of applied voltage. The transition was affected profoundly by the dispersed polymer network. The threshold was higher with shorter interpolymer network distance. Theoretically we studied the systems using a two-dimensional model in which the polymer networks were represented by parallel cylinders with random location. The interaction between the liquid crystal and the polymer network was described by the boundary condition imposed by the polymer network. By fitting the experimental data, we found that the polymer cylinders had diameters of a few submicrons, and a substantial amount of liquid crystal was trapped inside the cylinders. PMID- 11046439 TI - Curvature energy of a focal conic domain with arbitrary eccentricity AB - The most frequently observed focal conic domains (FCD's) in lamellar phases are those based on confocal paris of ellipse and hyperbola. Experimentally, the eccentricity of the ellipse takes a broad range of values 01 (under the constraint of a fixed major semiaxis of the ellipse); exceptions include situations with large saddle-splay elastic constant and small domains where the applicability of the elastic theory is limited. In realistic cases, a value of eccentricity smaller than 1 is stabilized by factors other than the curvature energy: by dislocations emerging from the FCD's with e not equal0, compression of layers and surface anchoring. PMID- 11046440 TI - Electroclinic liquid crystals with large induced tilt angle and small layer contraction AB - Optical and x-ray scattering studies of a chiral, organosiloxane smectic-A liquid crystal indicate a large field induced optical tilt of up to 31 degrees accompanied by a very small contraction of the smectic layers. This result suggests that the molecules have a nonzero tilt even with no applied field, and that the primary effect of the field is to induce long range order in the direction of the molecular tilt. PMID- 11046441 TI - Effect of optical purity on the critical heat capacity at the smectic-A-(chiral) smectic-C transition in an antiferroelectric liquid crystal AB - High resolution ac calorimetric measurements have been carried out near the smectic-A-chiral-smectic-C phase transition in the antiferroelectric liquid crystal 4-(1-methylheptyloxycarbonyl)phenyl 4(')-octyloxybiphenyl-4-carboxylate (MHPOBC). Data on samples with different optical purities have been analyzed in detail using a renormalization-group expression with corrections-to-scaling terms. The chiral-smectic-C(alpha)-chiral-smectic-C phase transition is first order, while the smectic-A-chiral-smectic-C(alpha) phase transition is second order. A direct smectic-A-chiral-smectic-C phase transition, which occurs in near racemic mixtures, was found to be quasitricritical and weakly first order. This implies that the smectic-A-smectic-C transition in the racemate locates at a special point where four critical lines intersect. PMID- 11046442 TI - Synchrotron x-ray study of the smectic layer directional instability AB - We have investigated the phenomenon of field-induced smectic layer instability, as monitored by synchrotron x-ray scattering. This instability means that, upon application of time-asymmetric electric fields to chiral smectics, the layer direction seems to "rotate" locally around an axis given by the direction of the applied field. For moderate values of field amplitude and asymmetry, domains with a favored layer inclination grow at the expense of unfavored ones, while larger fields and asymmetries generally lead to a chaotic flow behavior. At moderate amplitudes, we have followed the process of the horizontal layer folding (or horizontal chevron domain formation) and the smectic C* layer reorientation of ferroelectric liquid crystals by applying symmetric and asymmetric wave forms, respectively, and performing time resolved x-ray measurements. The studies unambiguously show the formation of a horizontal (in-plane, i.e., in a plane parallel to the cell substrates) chevron domain structure from a nonoriented sample by application of a symmetric electric field of sufficient amplitude. It is then demonstrated that a transition from the horizontal chevron domain structure to an in-plane uniform smectic layer direction takes place on application of asymmetric electric wave forms. Reversal of the field asymmetry reverses the inclination direction and selects the other layer normal direction as the uniform end state. The in-plane smectic layer reorientation process is followed here as it evolves, and analyzed directly by means of x-ray scattering. PMID- 11046443 TI - Statistical mechanics of vacancy and interstitial strings in hexagonal columnar crystals AB - Columnar crystals contain defects in the form of vacancy-interstitial loops or strings of vacancies and interstitials bounded by column "heads" and "tails." These defect strings are oriented by the columnar lattice and can change size and shape by movement of the ends and by forming kinks along the length. Hence an analysis in terms of directed living polymers [S. A. Safran, Statistical Thermodynamics of Surfaces, Interfaces, and Membranes (Addison-Wesley, Reading, MA, 1994), Sec. 8] is appropriate to study their size and shape distribution, volume fraction, etc. If the entropy of transverse fluctuations overcomes the string line tension in the crystalline phase, a string proliferation transition occurs, leading to a supersolid phase [E. Frey, D. R. Nelson, and D. S. Fisher, Phys. Rev. B 49, 9723 (1994); see also J. Prost, Liq. Cryst. 8, 123 (1990)]. We estimate the wandering entropy and examine the behavior in the transition regime. We also calculate numerically the line tension of various species of vacancies and interstitials in a triangular lattice for power-law potentials as well as for a modified Bessel function interaction between columns such as occurs in the case of flux lines in type-II superconductors or long polyelectrolytes in an ionic solution. We find that the centered interstitial is the lowest-energy defect for a very wide range of interactions; the symmetric vacancy is preferred only for extremely short interaction ranges. PMID- 11046444 TI - Effects of fluctuations in the orientational order parameter in the cyanobiphenyl (nCB) homologous series AB - Photopyroelectric measurements of the anisotropy in the thermal conductivity Deltak vs temperature in the nCB (n=5,ellipsis,9) series are reported. The data have been used to deduce the behavior of the orientational order parameter Q close to the nematic-isotropic (N-I) and smectic A-nematic (A-N) phase transitions, respectively. It has been shown that near the N-I transition the data for 5CB and 6CB are consistent with the so-called "tricritical hypothesis," which predicts beta=0.25. This is not true for 7CB and 8CB in which the order parameter exhibits a behavior that could be caused by the presence of fluctuations that become increasingly important when the transition temperature is approached. A very simple model, which takes into account the contribution of fluctuations to the orientational order, has been developed close to the A-N transition and it has been shown that it is in good agreement with the experimental results. A semiquantitative explanation for the observed behavior in compounds with different nematic range has been also given. PMID- 11046445 TI - Dynamic light scattering as a probe of orientational dynamics in confined liquid crystals AB - The eigenmodes of director orientational fluctuations in nematic liquid crystals in confined geometries were studied both theoretically and experimentally by dynamic light-scattering tehnique. The fundamental mode of the orientational fluctuations shows a crossover from bulk behavior, dominated by bulk elastic constant K, to surface dominated one, in which the relaxation rate is determined by the ratio of surface anchoring strength W and viscosity eta. The contribution of surface viscosity zeta is also significant when its characteristic length zeta/eta becomes comparable to the size of the confined system. It was measured in nematic liquid crystal in cylindrical pores of polycarbonate (Nuclepore) membranes to be of the order of 10 nm. PMID- 11046446 TI - Diffusion-limited aggregation: A relationship between surface thermodynamics and crystal morphology AB - We have combined the original diffusion-limited aggregation model introduced by Witten and Sander with the surface thermodynamics of the growing solid aggregate. The theory is based on the consideration of the surface chemical potential as a thermodynamic function of the temperature and nearest-neighbor configuration. The Monte Carlo simulations on a two-dimensional square lattice produce the broad range of shapes such as fractal dendritic structures, densely branching patterns, and compact aggregates. The morphology diagram illustrating the relationship between the model parameters and cluster geometry is presented and discussed. PMID- 11046447 TI - Nonlinear poisson-boltzmann theory of a wigner-seitz model for swollen clays AB - Swollen stacks of finite-size disclike Laponite clay platelets are investigated within a Wigner-Seitz cell model. Each cell is a cylinder containing a coaxial platelet at its center, together with an overall charge-neutral distribution of microscopic co and counterions, within a primitive model description. The nonlinear Poisson-Boltzmann (PB) equation for the electrostatic potential profile is solved numerically within a highly efficient Green's function formulation. Previous predictions of linearized Poisson-Boltzmann (LPB) theory are confirmed at a qualitative level, but large quantitative differences between PB and LPB theories are found at physically relevant values of the charge carried by the platelets. A hybrid theory treating edge effect at the linearized level yields good potential profiles. The force between two coaxial platelets, calculated within PB theory, is an order of magnitude smaller than predicted by LPB theory. PMID- 11046448 TI - Strong phase separation in a model of sedimenting lattices AB - We study the steady state resulting from instabilities in crystals driven through a dissipative medium, for instance, a colloidal crystal which is steadily sedimenting through a viscous fluid. The problem involves two coupled fields, the density and the tilt; the latter describes the orientation of the mass tensor with respect to the driving field. We map the problem to a one-dimensional lattice model with two coupled species of spins evolving through conserved dynamics. In the steady state of this model each of the two species shows macroscopic phase separation. This phase separation is robust and survives at all temperatures or noise levels- hence the term strong phase separation. This sort of phase separation can be understood in terms of barriers to remixing which grow with system size and result in a logarithmically slow approach to the steady state. In a particular symmetric limit, it is shown that the condition of detailed balance holds with a Hamiltonian which has infinite-ranged interactions, even though the initial model has only local dynamics. The long-ranged character of the interactions is responsible for phase separation, and for the fact that it persists at all temperatures. Possible experimental tests of the phenomenon are discussed. PMID- 11046449 TI - Hydrodynamic-flow-driven wetting in thin film polymer blends: growth kinetics and morphology AB - A thin film of deuterated poly(methyl methacrylate) (A) and poly(styrene-ran acrylonitrile) at the critical composition is annealed in the two phase region to induce simultaneous phase separation and wetting of the A-rich phase at the surface. Using forward recoil spectrometry, the wetting layer thickness is found to grow linearly with time at 185 degrees C and 190 degrees C. After selective etching of A, atomic force microscopy reveals a depletion layer having a bicontinuous, phase separated morphology. The A-rich tubes in this layer provide a pathway for rapid transport of the wetting phase from the bulk to the surface via hydrodynamic flow. Taken together, fast wetting layer growth t(1) and connectivity between the wetting layer and bulk provide unambiguous support for hydrodynamic-flow-driven wetting in thin film polymer blends. PMID- 11046450 TI - Phase diagram for morphological transitions of wetting films on chemically structured substrates AB - Using an interface displacement model we calculate the shapes of thin liquidlike films adsorbed on flat substrates containing a chemical stripe. We determine the entire phase diagram of morphological phase transitions in these films as function of temperature, undersaturation, and stripe width. PMID- 11046451 TI - Threshold criterion for wetting at the triple point AB - Grand canonical simulations are used to calculate adsorption isotherms of various classical gases on alkali metal and Mg surfaces. Ab initio adsorption potentials and Lennard-Jones gas-gas interactions are used. Depending on the system, the resulting behavior can be nonwetting for all temperatures studied, complete wetting, or (in the intermediate case) exhibit a wetting transition. An unusual variety of wetting transitions at the triple point is found in the case of a specific adsorption potential of intermediate strength. The general threshold for wetting near the triple point is found to be close to that predicted with a heuristic model of Cheng et al. This same conclusion was drawn in a recent experimental and simulation study of Ar on CO2 by Mistura et al. These results imply that a dimensionless wetting parameter w is useful for predicting whether wetting behavior is present at and above the triple temperature. The nonwetting/wetting crossover value found here is w approximately 3.3. PMID- 11046452 TI - Photon correlation spectroscopy: X rays versus visible light AB - We have performed combined dynamic light scattering (DLS) and dynamic x-ray scattering (DXS) experiments on dense colloidal suspensions. The intermediate scattering functions obtained with these two techniques are compared directly. In the case of optically index matched samples, the comparison demonstrates that DXS yields accurate and reliable results. It is shown that the hydrodynamic interaction H(q) can be determined experimentally, without taking recourse to any theoretical model, by combining DXS and DLS. The combination of the two methods probes the dynamics over more than one decade in scattering vector. Experiments on optically opaque samples, where DLS fails, demonstrate the necessity to use x rays in these systems. PMID- 11046453 TI - Potential energy landscape and long-time dynamics in a simple model glass AB - We analyze the properties of a Lennard-Jones system at the level of the potential energy landscape. After an exhaustive investigation of the topological features of the landscape of the systems, obtained by studying small size samples, we describe the dynamics of the systems in multidimensional configurational space by means of a simple model. This considers the configurational space as a connected network of minima where the dynamics proceeds by jumps described by an appropriate master equation. Using this model we are able to reproduce the long time dynamics and the low temperature regime. We investigate both the equilibrium regime and the off-equilibrium one, finding those typical glassy behaviors usually observed in the experiments such as (i) a stretched exponential relaxation, (ii) a temperature-dependent stretching parameter, (iii) a breakdown of the Stokes-Einstein relation, and (iv) the appearance of a critical temperature below which one observes a deviation from the fluctuation-dissipation relation as a consequence of the lack of equilibrium in the system. PMID- 11046454 TI - Phase diagram of highly asymmetric binary mixtures: A study of the role of attractive forces from the effective one-component approach AB - The phase diagram of an asymmetric solute-solvent mixture is investigated at the level of the effective one-component fluid. The solvent is taken into account by computing the potential of mean force between solute particles at infinite dilution for different models of solvent-solvent and solute-solvent short range interactions. Fluid-fluid and fluid-solid coexistence lines are determined from the free energy in the reference hypernetted chain theory for the fluid branch and from a variational perturbation theory for the solid one. The phase boundaries so determined compare well with recently published Monte Carlo data for mixtures of pure hard spheres. The influence of solute-solvent and solvent solvent short range attractive forces is then investigated. When compared with pure hard core interactions, these forces are found to produce dramatic changes in the phase diagram, especially on the solvent packing fractions at which a dense fluid of solutes can be stable and on the separation of the fluid-fluid and fluid-solid coexistence lines. Finally, the connection of these results with the behavior of some colloidal suspensions is emphasized. PMID- 11046455 TI - Flexible polymers and thin rods far from equilibrium: buckling dynamics AB - We investigate the dynamics of the classical Euler buckling instability of compressed objects such as flexible molecular chains and thin rods moving in a viscous medium. We find that flexible chains undergo a coarsening process self similar in time. They develop a wavelike pattern whose amplitude and wavelength grow in time. We relate the buckling dynamics to phase ordering phenomena. The role of the order parameter here is played by the chain slope with respect to the straight initial chain configuration. PMID- 11046456 TI - Self-diffusion in granular gases AB - The coefficient of self-diffusion for a homogeneously cooling granular gas changes significantly if the impact-velocity dependence of the restitution coefficient epsilon is taken into account. For the case of a constant epsilon the particles spread logarithmically slowly with time, whereas a velocity-dependent coefficient yields a power law time dependence. The impact of the difference in these time dependences on the properties of a freely cooling granular gas is discussed. PMID- 11046457 TI - Heterogeneous relaxation patterns in supercooled liquids studied by solvation dynamics AB - We have measured the solvation dynamics of a dipolar supercooled liquid near its glass transition in a temperature range in which the average structural relaxation time varies more than four orders of magnitude. The analysis of the time dependent average emission energy and the inhomogeneous linewidth of the S0< -T1(0-0) transition reveals that the orientation correlation decay pattern intrinsic in each relaxing unit is associated with a stretching exponent beta(intr)=1.00+/-0.08 in the entire range T(g)(KWW) the individual time constants remain correlated to their initial values at t=0. PMID- 11046458 TI - Replica field theory for a polymer in random media AB - In this paper we revisit the problem of a (non-self-avoiding) polymer chain in a random medium which was previously investigated by Edwards and Muthukumar (EM) [J. Chem. Phys. 89, 2435 (1988)]. As noticed by Cates and Ball (CB) [J. Phys. (France) 49, 2009 (1988)] there is a discrepancy between the predictions of the replica calculation of EM and the expectation that in an infinite medium the quenched and annealed results should coincide (for a chain that is free to move) and a long polymer should always collapse. CB argued that only in a finite volume one might see a "localization transition" (or crossover) from a stretched to a collapsed chain in three spatial dimensions. Here we carry out the replica calculation in the presence of an additional confining harmonic potential that mimics the effect of a finite volume. Using a variational scheme with five variational parameters we derive analytically for d<4 the result R approximately (g|ln &mgr;|)(-1/(4-d)) approximately (g ln V)(-1/(4-d)), where R is the radius of gyration, g is the strength of the disorder, &mgr; is the spring constant associated with the confining potential, and V is the associated effective volume of the system. Thus the EM result is recovered with their constant replaced by ln V as argued by CB. We see that in the strict infinite volume limit the polymer always collapses, but for finite volume a transition from a stretched to a collapsed form might be observed as a function of the strength of the disorder. For d<2 and for large V>V' approximately exp(g(2/(2-d))L((4-d)/(2-d))) the annealed results are recovered and R approximately (Lg)(1/(d-2)), where L is the length of the polymer. Hence the polymer also collapses in the large L limit. The one-step replica symmetry breaking solution is crucial for obtaining the above results. PMID- 11046459 TI - Glass transitions and dynamics in thin polymer films: dielectric relaxation of thin films of polystyrene AB - The glass transition temperature T(g) and the temperature T(alpha) corresponding to the peak in the dielectric loss due to the alpha process have been simultaneously determined as functions of film thickness d through dielectric measurements for polystyrene thin films supported on glass substrate. The dielectric loss peaks have also been investigated as functions of frequency for a given temperature. A decrease in T(g) was observed with decreasing film thickness, while T(alpha) was found to remain almost constant for d>d(c) and to decrease drastically with decreasing d for d>1 the percolation scaling of the diffusion coefficient D( perpendicular) approximately (deltaB/B(0))(0.7) is obtained. PMID- 11046481 TI - Experimental evidence of the effect of heat flux on thomson scattering off ion acoustic waves AB - Thomson self-scattering measurements are performed in a preionized helium gas jet plasma at different locations along the laser propagation direction. A systematic and important variation of the intensity ratio between the blue and the red ion spectral components is observed, depending on whether the location of the probed region is in front of or behind the focal plane. A simple theoretical calculation of Thomson scattering shows that this behavior can be qualitatively understood in terms of a deformation of the electron distribution function due to the return current correlated with the classical thermal heat flux. PMID- 11046482 TI - Optical mode structure of the plasma waveguide AB - The quasibound modes of an evolving plasma waveguide were investigated by using variably delayed end-injected and side-injected probe pulses. The use of these different coupling geometries allowed the probing of the waveguide's optical modes during two temporal regimes: early-time plasma channel development, characterized by leaky optical confinement, and later channel hydrodynamic expansion characterized by stronger confinement. The wave equation was solved to determine the available quasiguided optical modes and their confinement for experimentally measured electron density profiles. The guided intensity patterns and spectra measured at the waveguide exit were successfully explained in terms of these mode solutions. The spectrum of broadband end-coupled probe pulses was found to be unaffected by the guiding process, mainly because those modes which survived to the waveguide exit were well-bound, and for strongly bound fields, the transverse mode profiles are wavelength independent. By contrast, side coupling to the quasibound modes of the plasma waveguide was seen to be highly mode and frequency selective. PMID- 11046483 TI - Measurement of the radiative cooling rates for high-ionization species of krypton using an electron beam ion trap AB - We describe a measurement of the radiative cooling rate for krypton made at the Berlin electron beam ion trap (EBIT). The EBIT was tuned to a charge-state distribution approaching the ionization balance of a plasma at a temperature of about 5 keV. To determine the cooling rate, we made use of EBIT's capabilities to sample a wide range of electron-beam energies and distinguish between different radiation channels. We have measured the x-ray emission from bremsstrahlung, radiative recombination, dielectronic recombination, and line radiation following electron-impact excitation. The dominant contribution to the cooling rate is made by the n=3-2, n=4-2, ellipsis x rays of the L-shell spectra of krypton, which produce more than 75% of the total radiation loss. A difference with theoretical calculations is noted for the measured total cooling rate. The predicted values are lower by a factor of 1.5-2, depending on the theoretical model. For our measurement of the cooling rate, we estimate an uncertainty interval of 22-30 %. PMID- 11046484 TI - Shock compression of condensed matter using intense beams of energetic heavy ions AB - In this paper is presented, with the help of sophisticated two-dimensional hydrodynamic simulations, a suitable design with optimized parameters for a heavy ion beam-matter interaction experiment that will be carried out at the Gesellschaft fur Schwerionenforschung (GSI) Darmstadt by the end of the year 2001 when the upgrade of the existing accelerator facility will be completed. Our simulations show that this upgraded heavy-ion beam is capable of generating strong shocks in solid targets that compress the target material to supersolid densities and generate multi-mbar pressures. This will open up, at the GSI, the possibility of investigation of the equation-of-state properties of matter under such extreme conditions. Numerical simulations can predict the experimental results with reasonable accuracy, which is helpful in designing the diagnostic tools for the experiment. PMID- 11046485 TI - Conditions for electron capture by an ultraintense stationary laser beam AB - We present in this paper a quantitative study of an effect, in which a low-energy free electron is captured and violently accelerated to GeV final kinetic energy by a stationary extra-high-intensity laser beam (Q0 identical witheE/m(e)omegac greater, similar100). The conditions under which this phenomenon can occur, such as the momentum range, incident angle of the incoming electron, the waist width of the laser beam, etc., have been investigated in detail. PMID- 11046486 TI - Transition from amplified spontaneous emission to laser action in strongly scattering media AB - In an active random medium, the combination of multiple scattering with light amplification may lead to random laser action. However, it is crucial but sometimes difficult to distinguish between amplified spontaneous emission and lasing. By varying the amount of scattering in an amplifying random medium, we have observed the transition from amplified spontaneous emission to lasing with coherent feedback. We have found out when the transition occurs through the measurement of the scattering mean free path. Our numerical simulation based on the direct solution to Maxwell equations clearly illustrates the transition from light amplification to laser oscillation due to an increase of the amount of scattering in active random medium. PMID- 11046487 TI - Exact description of photon migration in anisotropically scattering media AB - The aim of the present paper is to deliver a method for exact calculation of the probability of photon migration in an infinite and homogeneous medium scattering photons anisotropically. The phase function is represented as an expansion over spherical harmonics. The probability of photon migration is obtained as an expansion with respect to the number of scatterings with the coefficients depending on the distance-to-time ratio and can be easily calculated from recurrence relations. Up to 30 scatterings are taken into account when computing the migration probabilities, the number of effectively contributing scatterings being strongly dependent on the distance-to-time ratio decreasing when one approaches the propagation front. The important property of the method is its capability to describe exactly migration from the early arriving photons up to the scattering ones. The limits of the approximate description of anisotropic scattering as isotropic with an effective value of the scattering coefficient is analyzed by calculating the best-fit value of the scattering coefficient. PMID- 11046488 TI - Stabilization of the kerr effect by self-induced ionization: formation of optical light spatially localized structures AB - The nonlinear propagation of ultrashort laser pulses launched into the air is investigated. The formation of optical light "bullets," or spatially localized structures, has been experimentally observed recently. Their stability is shown as due to the occurrence of a dynamical balance between two opposite nonlinear effects: an optical focusing Kerr effect balanced by a defocusing self-induced multiphoton partial ionization of the neutral gas. Characteristics of the "bullets" are predicted analytically and confirmed numerically. They are found to be in agreement with observations. PMID- 11046489 TI - Two-dimensional discrete breathers: construction, stability, and bifurcations AB - We develop a methodology for the construction of two-dimensional discrete breather excitations. Application to the discrete nonlinear Schrodinger equation on a square lattice reveals three different types of breathers. Considering an elementary plaquette, the most unstable mode is centered on the plaquette, the most stable mode is centered on its vertices, while the intermediate (but also unstable) mode is centered at the middle of one of the edges. Below the turning points of each branch in a frequency-power phase diagram, the construction methodology fails and a continuation method is used to obtain the unstable branches of the solutions until a triple point is reached. At this triple point, the branches meet and subsequently bifurcate into the final state of an extended phonon mode. PMID- 11046491 TI - Coefficient of restitution for one-dimensional harmonic solids AB - Using a numerical algorithm based on the time evolution of normal modes, we calculate the coefficient of restitution eta for various one-dimensional harmonic solids colliding with a hard wall. We find that, for a homogeneous chain, eta=1 in the thermodynamic limit. However, for a chain in which weaker springs are introduced in the colliding front half, eta remains significantly less than one even in the thermodynamic limit, and the "lost" energy goes mostly into low frequency normal modes. An understanding of these results is given in terms of how the energy is redistributed among the normal modes as the chain collides with the wall. We then contrast these results with those for collisions of one dimensional harmonic solids with a soft wall. Using perturbation theory, we find that eta=1 for all harmonic chains in the extremely soft wall limit, but that inelasticity grows with increasing chain size in contrast to hard wall collisions. PMID- 11046490 TI - Analytical solution for photorefractive screening solitons AB - We study formation and interaction of one-dimensional screening solitons in a photorefractive medium with sublinear dependence of the photoconductivity on light intensity. We find an exact analytical solution to the corresponding nonlinear Schrodinger equation. We show that these solitons are stable in propagation and their interaction is generic for solitons of saturable nonlinearity. In particular, they may fuse or "give birth" to new solitons upon collision. PMID- 11046492 TI - Parameters characterizing electromagnetic wave polarization AB - In this paper, generalizations of the Stokes parameters and alternative characterizations of three-dimensional (3D) time-varying electromagnetic fields is introduced. One of these characteristics is the normal of the polarization plane, which, in many cases of interest, is parallel (or antiparallel) to the direction of propagation. Others are the two spectral density Stokes parameters which describe spectral intensity and circular polarization. The analysis is based on the spectral density tensor. This tensor is expanded in a base composed of the generators of the SU(3) symmetry group, as given by Gell-Mann and Y. Ne'eman [The Eight-fold Way (Benjamin, New York, 1964)] and the coefficients of this expansion are identified as generalized spectral density polarization parameters. The generators have the advantage that they obey the same algebra as the Pauli spin matrices, which is the base for expanding the 2D spectral density tensor with the Stokes parameters as coefficients. The polarization parameters introduced are formulated in the frequency domain, thereby further generalizing the theory to allow for wide-band electromagnetic waves in contrast to the traditional quasi-monochromatic formulation. PMID- 11046493 TI - Shaping of photorefractive two-wave coupling by fast phase modulation AB - Using the distinctive features of the photorefractive nonlinearity, we derive a general self-consistent set of equations to describe two-wave coupling in the presence of fast and arbitrary strong-phase modulation. By considering a number of important particular cases, we show that phase modulation is a powerful and useful tool for shaping the characteristics of two-wave coupling such as the value of the energy exchange, the diffraction efficiency of the recorded grating, and the structure of the grating fringes. Finally, we analyze the role of the phase modulation in the active stabilization of wave coupling by means of an electronically introduced phase feedback. PMID- 11046494 TI - Second-harmonic generation of the nth-order Bessel beam. AB - We investigate the second-harmonic generation of the nth-order Bessel beam in the nonlinear medium. The analysis is based on the Khokhlov-Zabolotskaya-Kuznetsov wave equation under the second-order approximation in nonlinear acoustics. The theory indicates that for an nth-order Bessel beam, the second-harmonic beam is nearly diffraction-free in the radial direction and behaves as a Bessel beam of the order 2n, and that the axial pressure amplitude is proportional to the square root of propagation distance. A variety of applications in many fields of nonlinear acoustics and nonlinear optics is expected. PMID- 11046495 TI - Optical vortex solitons in parametric wave mixing AB - We analyze two-component spatial optical vortex solitons supported by parametric wave mixing processes in a nonlinear bulk medium. We study two distinct cases of such localized waves, namely, parametric vortex solitons due to phase-matched second-harmonic generation in an optical medium with competing quadratic and cubic nonlinear response, and vortex solitons in the presence of third-harmonic generation in a cubic medium. We find, analytically and numerically, the structure of two-component vortex solitons, and also investigate modulational instability of their plane-wave background. In particular, we predict and analyze in detail novel types of vortex solitons, a "halo-vortex," consisting of a two component vortex core surrounded by a bright ring of its harmonic field, and a "ring-vortex" soliton which is a vortex in a harmonic field that guides a ring like localized mode of the fundamental-frequency field. PMID- 11046496 TI - Time step bias improvement in diffusion monte carlo simulations AB - A Makri-Miller approximation to the exact propagator and the improved split operator propagator proposed by Drozdov are implemented within the diffusion Monte Carlo method for the simulation of boson systems, and confronted with the Trotter formula and with the importance sampling technique. As a preliminary approach, we compute analytically the time step bias of the mean energy for the different propagators in the simple case of the harmonic oscillator. These results indicate the improved split-operator propagator as the most accurate. Simulations on one- and three-dimensional model systems confirm the analytical results showing that this propagator is very efficient in reducing the time step bias, therefore improving the efficiency of the algorithm. PMID- 11046497 TI - Optimized energy calculation in lattice systems with long-range interactions AB - We discuss an efficient approach to the calculation of the internal energy in numerical simulations of spin systems with long-range interactions. Although, since the introduction of the Luijten-Blote algorithm, Monte Carlo simulations of these systems no longer pose a fundamental problem, the energy calculation is still an O(N2) problem for systems of size N. We show how this can be reduced to an O(N log N) problem, with a break-even point that is already reached for very small systems. This allows the study of a variety of, until now hardly accessible, physical aspects of these systems. In particular, we combine the optimized energy calculation with histogram interpolation methods to investigate the specific heat of the Ising model and the first-order regime of the three state Potts model with long-range interactions. PMID- 11046498 TI - Treatment of laser-induced thermal acoustics in the framework of discrete kinetic theory AB - The physics behind the laser-induced thermal acoustics technique is dealt with on a microscopic level. A discrete velocity model of the Boltzmann equation for inelastically interacting gas mixtures in the presence of two counterpropagating laser beams is established. The collisional scheme for the model is developed by taking into account elastic and inelastic interactions between the gas particles, on the one hand, and the interactions between monochromatic laser photons and gas particles, on the other hand. The formation and evolution of laser-pulse-driven thermal and density gratings are simulated by numerically solving the discrete kinetic equations based on the fractional step method. Numerical results are provided for a wide scope of Knudsen numbers. PMID- 11046499 TI - Solving the bound-state Schrodinger equation by reproducing kernel interpolation AB - Based on reproducing kernel Hilbert space theory and radial basis approximation theory, a grid method is developed for numerically solving the N-dimensional bound-state Schrodinger equation. Central to the method is the construction of an appropriate bounded reproducing kernel (RK) Lambda(alpha)( ||r ||) from the linear operator -nabla(2)(r)+lambda(2) where nabla(2)(r) is the N-dimensional Laplacian, lambda>0 is a parameter related to the binding energy of the system under study, and the real number alpha>N. The proposed (Sobolev) RK Lambda(alpha)(r,r(')) is shown to be a positive-definite radial basis function, and it matches the asymptotic solutions of the bound-state Schrodinger equation. Numerical tests for the one-dimensional (1D) Morse potential and 2D Henon-Heiles potential reveal that the method can accurately and efficiently yield all the energy levels up to the dissociation limit. Comparisons are also made with the results based on the distributed Gaussian basis method in the 1D case as well as the distributed approximating functional method in both 1D and 2D cases. PMID- 11046500 TI - Coupled nonequilibrium growth equations: self-consistent mode coupling using vertex renormalization AB - We find that studying the simplest of the coupled nonequilibrium growth equations of Barabasi by self-consistent mode coupling requires the use of dressed vertices. Using the vertex renormalization, we find a roughening exponent which already in the leading order is quite close to the numerical value. PMID- 11046501 TI - Ratio of canonical and microcanonical temperatures of a vibratory antiferromagnetic ising chain AB - The ratio of canonical and microcanonical temperatures T(c)/T(&mgr;) of a vibratory antiferromagnetic Ising chain with N spins is given by analytical calculation. The result is T(c)/T(&mgr;)=1+O(N-1), which is consistent with the natural assumption given by Rugh. PMID- 11046502 TI - Large deviation function for the eden model and universality within the one dimensional kardar-parisi-zhang class AB - It has been recently conjectured that for large systems, the shape of the central part of the large deviation function of the growth velocity would be universal for all the growth systems described by the Kardar-Parisi-Zhang equation in 1+1 dimension. One signature of this universality would be that the ratio of cumulants R(t)=[(c)](2)/[(c)(c)] would tend towards a universal value 0.415 17ellipsis as t tends to infinity, provided periodic boundary conditions are used. This has recently been questioned by Stauffer. In this paper we summarize various numerical and analytical results supporting this conjecture, and report in particular some numerical measurements of the ratio R(t) for the Eden model. PMID- 11046503 TI - Coherent structure analysis of spatiotemporal chaos AB - We introduce a measure to quantify spatiotemporal turbulence in extended systems. It is based on the statistical analysis of a coherent structure decomposition of the evolving system. Applied to a cellular excitable medium and a reaction diffusion model describing the oxidation of CO on Pt(100), it reveals power-law scaling of the size distribution of coherent space-time structures for the state of spiral turbulence. The coherent structure decomposition is also used to define an entropy measure, which sharply increases in these systems at the transition to turbulence. PMID- 11046504 TI - Ground state of a dipolar crystal AB - We provide some of the strongest evidence to date that the ground state structure of an infinite collection of point dipoles with hardcore sphere interactions is body-centered tetragonal. The structure with the next highest binding energy is not face-centered cubic; a particular honeycomb structure has lower energy. PMID- 11046505 TI - Higher-order dynamics in lattice-based models using the chapman-enskog method AB - In this paper, we investigate the existence of higher-order dynamics in lattice based models. We have identified two conditions that determine whether a model would allow some Burnett-like equations when the Chapman-Enskog expansion is used. These two conditions are the number of the conserved quantity as well as the space and time discretization. We shall demonstrate these conditions by discussing (1) pure diffusion equation and (2) hydrodynamic equations. While the fact that diffusion equation allows the higher-order dynamics can be shown easily, we will illustrate that care must be taken when deriving Burnett-like equations for lattice-based hydrodynamics models using the Chapman-Enskog method. PMID- 11046506 TI - Structure of vortices in a karman street behind a heated cylinder AB - The velocity and temperature fields in a wake behind a heated cylinder are investigated for Reynolds numbers in the interval 65 approximately t(2nu) and the average number of visited sites approximately t(k), are universal and given by nu=1/(d+1) and k=d/(d+1). Below u(c), the walk swells and the exponents are as with no interaction, i.e., nu=1/2 for all d, k=1/2 for d=1 and k=1 for d>/=2. At u(c), the exponents are found to be in a different universality class. PMID- 11046526 TI - Stabilizing nonlinear dynamical systems by an adaptive adjustment mechanism AB - An adaptive adjustment mechanism is applied to stabilize multidimensional dynamical systems. Without utilizing any prior knowledge of the system itself, nor extra external control signals, the mechanism can ensure a large class of chaotic systems to converge to their "generic" stable periodic orbits. PMID- 11046525 TI - Chiral-glass transition and replica symmetry breaking of a three-dimensional heisenberg spin glass AB - Extensive equilibrium Monte Carlo simulations are performed for a three dimensional Heisenberg spin glass with the nearest-neighbor Gaussian coupling to investigate its spin-glass and chiral-glass orderings. The occurrence of a finite temperature chiral-glass transition without the conventional spin-glass order is established. Critical exponents characterizing the transition are different from those of the standard Ising spin glass. The calculated overlap distribution suggests the appearance of a peculiar type of replica-symmetry breaking in the chiral-glass ordered state. PMID- 11046527 TI - Relationships between a roller and a dynamic pressure distribution in circular hydraulic jumps AB - We investigated numerically the relation between a roller and the pressure distribution to clarify the dynamics of the roller in circular hydraulic jumps. We found that a roller which characterizes a type II jump is associated with two high pressure regions after the jump, while a type I jump (without the roller) is associated with only one high pressure region. Our numerical results show that building up an appropriate pressure field is essential for a roller. PMID- 11046528 TI - Time correlated single photon mie scattering from a sonoluminescing bubble AB - Application of time correlated single photon counting to pulsed Mie scattering enables one to resolve changes in light scattering to better than 50 ps. This technique is applied to the highly nonlinear motion of a sonoluminescing bubble. Physical processes, such as outgoing shock wave emission, that limit the interpretation of the data are measured with a streak camera and microscopy. Shock speeds about 6 km/s have been observed. PMID- 11046529 TI - Large deviation statistics of the energy-flux fluctuation in the shell model of turbulence AB - The energy-flux fluctuation in the shell model of turbulence is numerically analyzed from the large deviation statistical point of view. We first observe that the rate function defined in the inertial range is independent of the Reynolds number. The rate function derived by the cascade model of the log Poisson statistics turns out to be in good agreement with the present numerical result in the region where strong singularity of fluctuation exits. This fact may imply the universality as well as the robustness of the large deviation statistical quantities in turbulence. PMID- 11046530 TI - Crystallization and phase separation in nonadditive binary hard-sphere mixtures AB - We calculate for the first time the full phase diagram of an asymmetric nonadditivehard-sphere mixture. The nonadditivity strongly affects the crystallization and the fluid-fluid phase separation. The global topology of the phase diagram is controlled by an effective size ratio y(eff), while the fluid solid coexistence scales with the depth of the effective potential well. PMID- 11046531 TI - First-order phase transition in a (1+1)-dimensional nonequilibrium wetting process AB - A model for nonequilibrium wetting in 1+1 dimensions is introduced. It comprises adsorption and desorption processes with a dynamics that generically does not obey detailed balance. Depending on the rates of the dynamical processes the wetting transition is either of first or second order. It is found that the wet (unbound) and the nonwet (pinned) states coexist and are both thermodynamically stable in a domain of the dynamical parameters that define the model. This is in contrast with equilibrium transitions where coexistence of thermodynamically stable states takes place only on the transition line. PMID- 11046532 TI - Evidence for dipole surface orientational order at critical interfaces AB - At the critical interface of dipolar systems theory predicts that the amplitude of the surface orientational order alpha(2)(z) approximately m(*4)d(2)v(z)/dz(2), where m(*) is a reduced dipole moment and v(z) is the local composition at position z within the interface. We find quantitative agreement with these predictions for two different critical binary liquid mixtures composed of a highly polar and a nonpolar component. PMID- 11046533 TI - Glassy dynamics of protein folding. AB - A coarse-grained model of a random polypeptide chain, with only discrete torsional degrees of freedom and Hookean springs connecting pairs of hydrophobic residues is shown to display stretched exponential relaxation under Metropolis dynamics at low temperatures with the exponent beta approximately 1/4, in agreement with the best experimental results. The time dependent correlation functions for fluctuations about the native state, computed in the Gaussian approximation for real proteins, have also been found to have the same functional form. Our results indicate that the energy landscape exhibits universal features over a very large range of energies and is relatively independent of the specific dynamics. PMID- 11046534 TI - Solitons in anharmonic chains with ultra-long-range interatomic interactions AB - We study the influence of long-range interatomic interactions on the properties of supersonic pulse solitons in anharmonic chains. We show that in the case of ultra-long-range (e.g., screened Coulomb) interactions three different types of pulse solitons coexist in a certain velocity interval: one type is unstable but the two others are stable. The high-energy stable soliton is broad and can be described in the quasicontinuum approximation. But the low-energy stable soliton consists of two components, short-range and long-range ones, and can be considered as a bound state of these components. PMID- 11046535 TI - Self-similarity and fractals in soliton-supporting systems AB - We describe a principle that can be used to generate self-similarity and fractals in almost any nonlinear system in nature that supports solitons, given that some proper nonadiabatic conditions are met. We illustrate our idea on a particular optics example that also theoretically demonstrates fractals in nonlinear optics. PMID- 11046596 TI - Magnetic resonance imaging of convection in laser-polarized xenon. AB - We demonstrate nuclear magnetic resonance (NMR) imaging of the flow and diffusion of laser-polarized xenon (129Xe) gas undergoing convection above evaporating laser-polarized liquid xenon. The large xenon NMR signal provided by the laser polarization technique allows more rapid imaging than one can achieve with thermally polarized gas-liquid systems, permitting shorter time-scale events such as rapid gas flow and gas-liquid dynamics to be observed. Two-dimensional velocity-encoded imaging shows convective gas flow above the evaporating liquid xenon, and also permits the measurement of enhanced gas diffusion near regions of large velocity variation. PMID- 11046627 TI - Synchronization due to common pulsed input in Stein's model. AB - It is known that stimulus-evoked oscillatory synchronization among neurones occurs in widely separated cortical regions. In this paper we provide a possible mechanism to explain the phenomenon. When a common, random input is presented, we find that a group of neurones-of Stein's (integrate-and-fire) model type with or without reversal potentials-are capable of quickly synchronizing their firing. Interestingly the optimal average synchronization time occurs when the common input has a high coefficient of variation of interspike intervals (greater than 0.5) for this model with or without reversal potentials. The model with reversal potentials more quickly synchronizes than that without reversal potentials. PMID- 11046628 TI - Error propagation in the hypercycle. AB - We study analytically the steady-state regime of a network of n error-prone self replicating templates forming an asymmetric hypercycle and its error tail. We show that the existence of a master template with a higher noncatalyzed self replicative productivity a than the error tail ensures the stability of chains in which m < n-1 templates coexist with the master species. The stability of these chains against the error tail is guaranteed for catalytic coupling strengths K on the order of a. We find that the hypercycle becomes more stable than the chains only if K is on the order of a2. Furthermore, we show that the minimal replication accuracy per template needed to maintain the hypercycle, the so called error threshold, vanishes as square root of n/K for large K and N < or = 4. PMID- 11046629 TI - Small-angle neutron scattering from large unilamellar vesicles: an improved method for membrane thickness determination. AB - Small-angle neutron scattering (SANS) measurements were performed on large unilamellar vesicles (LUVs) in order to investigate solute effects on membrane properties. Although SANS is a well established technique for the measurement of membrane thickness in unilamellar vesicles, earlier measurements have depended on approximate treatments of the scattering function and have suffered from effects of multilamellarity or difficulty in sample preparation. More recent studies of temperature induced thickness changes in DPPC LUVs which have included explicit treatment of the full scattering function were complicated by disparities between the predicted and measured scattering curves. Here, we reexamine theoretical descriptions of SANS from LUVs. Motivated by our observations, we then introduce a new method for interpretation of SANS data, which we compare to established techniques and apply to our measurements. PMID- 11046630 TI - Multiplicity of species in some replicative systems. AB - In an attempt to explain the uniqueness of the coding mechanism of living cells as contrasted with the multispecies structure of ecosystems we examine two models of individuals with some replicative properties. In the first model the system generically remains in a multispecies state. Even though for some of these species the replicative probability is very high, they are unable to invade the system. In the second model, in which the death rate depends on the type of the species, the system relatively quickly reaches a single-species state and fluctuations might at most bring it to yet another single-species state. PMID- 11046683 TI - Simulated coevolution in a mutating ecology. AB - The bit-string Penna model is used to simulate the competition between an asexual parthenogenetic and a sexual population sharing the same environment. A newborn of either population can mutate and become a part of the other with some probability. In a stable environment the sexual population soon dies out. When an infestation by rapidly mutation genetically coupled parasites is introduced, however, sexual reproduction prevails, as predicted by the so-called Red Queen hypothesis for the evolution of sex. PMID- 11046684 TI - Scale-invariant fluctuations at different levels of organization in developing heart cell networks. AB - In the last few years it has been realized that the intervals of spontaneous spiking events in the intact heart exhibit coexisting scale-invariant count fluctuations and anticorrelated interspike intervals fluctuations. Here, we show experimentally that this feature is an intrinsic property of single isolated heart cells, which is preserved when the cells couple into networks. We present a model explaining this behavior at both the single cell and network levels. PMID- 11047755 TI - Mitochondrial encephalomyopathy and complex III deficiency associated with a stop codon mutation in the cytochrome b gene. AB - We have reinvestigated a young woman, originally reported by us in 1983, who presented with exercise intolerance and lactic acidosis associated with severe deficiency of complex III and who responded to therapy with menadione and ascorbate. Gradually, she developed symptoms of a mitochondrial encephalomyopathy. Immunocytochemistry of serial sections of muscle showed a mosaic of fibers that reacted poorly with antibodies to subunits of complex III but reacted normally with antibodies to subunits of complexes I, II, or IV, suggesting a mutation of mtDNA. These findings demonstrate the diagnostic value of immunocytochemistry in identifying specific respiratory-chain deficiencies and, potentially, distinguishing between nuclear- or mtDNA-encoded defects. Sequence analysis revealed a stop-codon mutation (G15242A) in the mtDNA-encoded cytochrome b gene, resulting in loss of the last 215 amino acids of cytochrome b. PCR-RFLP analysis indicated that the G15242A mutation was heteroplasmic and was present in a high percentage (87%) of affected tissue (skeletal muscle) and a low percentage (0.7%) of unaffected tissue (blood) but was not detected in controls. Analysis of microdissected muscle fibers showed a significant correlation between the immunoreactivity toward the Rieske protein of complex III and the percentage of mutant mtDNA: immunopositive fibers had a median value of 33% of the G15242A mutation, whereas immunonegative, ragged-red fibers had a median value of 89%, indicating that the stop-codon mutation was pathogenic in this patient. The G15242A mutation was also present in several other tissues, including hair roots, indicating that it must have arisen either very early in embryogenesis, before separation of the primary germ layers, or in the maternal germ line. The findings in this patient are contrasted with other recently described patients who have mutations in the cytochrome b gene. PMID- 11047756 TI - Identification of a locus for autosomal dominant polycystic liver disease, on chromosome 19p13.2-13.1. AB - Polycystic liver disease (PCLD) is characterized by the growth of fluid-filled cysts of biliary epithelial origin in the liver. Although the disease is often asymptomatic, it can, when severe, lead to complications requiring surgical therapy. PCLD is most often associated with autosomal dominant polycystic kidney disease (ADPKD); however, families with an isolated polycystic liver phenotype without kidney involvement have been described. The clinical presentation and histological features of polycystic liver disease in the presence or absence of ADPKD are indistinguishable, raising the possibility that the pathogenetic mechanisms in the diseases are interrelated. We ascertained two large families with polycystic liver disease without kidney cysts and performed a genomewide scan for genetic linkage. A causative gene, PCLD, was mapped to chromosome 19p13.2-13.1, with a maximum LOD score of 10.3. Haplotype analysis refined the PCLD interval to 12.5 cM flanked by D19S586/D19S583 and D19S593/D19S579. The discovery of genetic linkage will facilitate diagnosis and study of this underdiagnosed disease entity. Identification of PCLD will be instrumental to an understanding of the pathogenesis of cyst formation in the liver in isolated PCLD and in ADPKD. PMID- 11048268 TI - Historical survey of laser dentistry. AB - Soon after lasers were invented, investigators began to examine the effects of different wavelengths of laser energy on oral tissues. From soft tissue surgery to restorative dentistry, from reshaping healthy gingiva to treating pathologic conditions, from routine procedures to experimental applications, researchers and clinicians have defined the limitations and shown the advantages of laser use in dentistry. PMID- 11047757 TI - A novel X-linked disorder of immune deficiency and hypohidrotic ectodermal dysplasia is allelic to incontinentia pigmenti and due to mutations in IKK-gamma (NEMO). AB - Hypohidrotic ectodermal dysplasia (HED), a congenital disorder of teeth, hair, and eccrine sweat glands, is usually inherited as an X-linked recessive trait, although rarer autosomal dominant and recessive forms exist. We have studied males from four families with HED and immunodeficiency (HED-ID), in which the disorder segregates as an X-linked recessive trait. Affected males manifest dysgammaglobulinemia and, despite therapy, have significant morbidity and mortality from recurrent infections. Recently, mutations in IKK-gamma (NEMO) have been shown to cause familial incontinentia pigmenti (IP). Unlike HED-ID, IP affects females and, with few exceptions, causes male prenatal lethality. IKK gamma is required for the activation of the transcription factor known as "nuclear factor kappa B" and plays an important role in T and B cell function. We hypothesize that "milder" mutations at this locus may cause HED-ID. In all four families, sequence analysis reveals exon 10 mutations affecting the carboxy terminal end of the IKK-gamma protein, a domain believed to connect the IKK signalsome complex to upstream activators. The findings define a new X-linked recessive immunodeficiency syndrome, distinct from other types of HED and immunodeficiency syndromes. The data provide further evidence that the development of ectodermal appendages is mediated through a tumor necrosis factor/tumor necrosis factor receptor-like signaling pathway, with the IKK signalsome complex playing a significant role. PMID- 11048269 TI - An overview of laser wavelengths used in dentistry. AB - The scientific basis and tissue effects of dental lasers have been discussed. It is most important for the dental practitioner to become familiar with those principles, then choose the proper lasers for the intended clinical application. Although there is some overlap of the type of tissue interaction, each wavelength has specific qualities that accomplish a specific treatment objective. Other articles in this issue provide an in-depth study of the uses of lasers in dentistry. PMID- 11048270 TI - Laser use in fixed, removable, and implant dentistry. AB - Laser use in reconstructive dental procedures (fixed, removable, or implant) provides a more controlled environment at the operative site. Control is a prerequisite for successful prosthetics and is accomplished by the unique ability of the laser to interact photo/thermally with biologic tissues using the lightest touch or none at all. Hemostasis is the result of this interaction by coagulating and sealing small blood and lymph vessels and vaporizing soft tissue. This hemostasis allows for more exact soft tissue contours to replicate or improve the periodontium before reconstruction, whether fixed or removable. The biologic seal is a unique phenomenon that occurs in the jaws of periimplant tissues of patients as a result of implant treatment. The biologic seal is paramount to the longevity and service of the implant. Use of dental laser technology provides the advanced laser and reconstructive dentist with the tool to improve the seal and improve the prognosis of the implant. The seal must be maintained by good oral hygiene and prophylactic procedures, however. PMID- 11048271 TI - Use of lasers in periodontics. AB - Although lasers have been used in periodontal therapy for 10 or more years, clinicians have barely begun to tap the enormous potential of this form of therapy. The relative lack of pain, ease of use, and site specificity of the laser make it an ideal addition to the periodontist's armamentarium. Applications now are being developed for a broader range of wavelengths that will offer useful, predictable, and comfortable therapy for managing the periodontal patient. PMID- 11048272 TI - Laser de-epithelialization for enhanced guided tissue regeneration. A paradigm shift? AB - The rationale for laser de-epithelialization stems from the attempts to block the down-growth of epithelium into the healing periodontal wound after surgery and prevent formation of a long junctional epithelial attachment. This concept has seen numerous techniques for accomplishing the blockage of epithelium. The advent of GTR was an offshoot of this concept and led Gottlow et al [table: see text] to examine the effects of selectively blocking certain cell types from contacting the root surface during periodontal wound healing. The use of a CO2 laser to de epithelialize the gingival flaps is an attempt to exclude this cell type from the healing wound and has been used with and without the benefit of GTR membranes. In a study on beagle dogs, the histologic results of using membranes and the laser procedure enhanced the wound healing and regeneration of new bone, cementum, and connective tissue attachment when compared with paired defects using the membranes alone. The results from the human studies and case reports combined with the animal studies indicate a positive benefit in wound healing because of the laser de-epithelialization technique. The use of an osseous graft in treatment of periodontal defects has been shown to stimulate new bone growth effectively and to regenerate new attachment. It has been speculated that the additional benefit of an osseous graft in GTR procedures is the organization of the blood clot at initial healing, which may tend to maintain the space needed for regeneration and to provide a matrix for the fibrin clot to retard epithelial down-growth. Studies comparing the results of osseous grafting with flap debridement always have shown that the amount of new bone formation and clinical new attachment favor the grafted sites versus paired nongrafted sites. The effects of removal of the pocket epithelium at the time of periodontal surgery have been studied by several authors, and these studies generally shown an incomplete removal of the sulcular epithelium by the inverse bevel incision. Epithelial excision was studied by Centty et al, who compared the removal of sulcular epithelium by the CO2 laser technique with conventional methods. Their results confirm that (1) a more complete removal of sulcular epithelium was obtained by laser than by knives, and (2) the technique effectively removes the oral and sulcular epithelium from a gingival flap without damaging the viability of the flap during wound healing. The technique as described in this article was used by Israel et al to verify further the ability to maintain a viable gingival flap during multiple laser deepithelialization procedures in humans during the first 30 days of healing. [table: see text] The concept of laser de epithelialization as an adjunct to regenerative periodontal procedures currently is being studied in a multicenter university setting using a parallel study in controlled clinical trials. The first of these reports was mentioned previously (Araujo et al, unpublished data) and shows the enhanced wound healing of periodontal defects through use of the laser de-epithelialization technique. The authors believe that this technique has shown significantly better results than those obtained through conventional osseous grafting alone and appears to be comparable to the results reported for GTR procedures with barrier membranes. This concept provides a paradigm shift from the conventional use of GTR therapy by acknowledging the difficulty in controlling epithelium during the early wound healing. It also allows a more comprehensive therapy for treating periodontal disease that addresses the generalized nature of the disease, with multiple lesions being treated concurrently in an economical manner. The patient presenting with generalized advanced periodontal disease could have several defects definitively treated in one quadrant using the laser deepithelialization technique without the need for multiple membrane therapy. (ABSTRACT TRUNCATE PMID- 11048273 TI - Integration of lasers into a soft tissue management program. AB - The future of laser use in periodontal procedures is bright. As techniques are improved and devices are refined further, lasers will become an important part of the armamentarium in laser treatment. Research is currently underway using lasers in excisional new attachment procedures for periodontal ligament and bone regeneration procedures. Although much scientific work needs to be done in this area, the results appear promising. The Nd:YAG laser is an excellent addition to the armamentarium of the general practitioner in the treatment of early to moderate periodontal disease. When using proper diagnostic and treatment planning protocols and with proper energy settings and technique, the laser elevates the level of care that can be provided to patients in a conservative fashion. It is an instrument that achieves maximum oral health in a minimally invasive fashion. PMID- 11048274 TI - Lasers in pediatric and adolescent dentistry. AB - The advent of lasers has offered new possibilities for improved service for pediatric dentistry patients and their parents. The removal of tissues without anesthesia is a common benefit. Almost symptomless postoperative experiences are in marked contrast to conventional methods. Whether performing cavity preparations or remodeling gingival architecture, the acceptance from a young patient is a gratification as well as a practice builder. Because many children may experience laser treatment as their first contact with dentistry, there is a possibility that a new generation of patients will grow up with a different attitude toward dentistry. Parents are enthusiastic about being able to offer their children the advantages of laser care. PMID- 11048276 TI - Lasers in oral and maxillofacial surgery. AB - Because of their many advantages, lasers have become indispensable in OMS as a modality for soft tissue surgery. Based on manufacturer estimates, approximately 10% to 20% of all oral and maxillofacial surgeons have one or more lasers in their offices, and most surgeons have access to lasers in the hospital. Lasers not only enhance the current surgical options for treatment, but also have expanded the scope of practice. There are many uses for lasers in OMS, and the advent of new wavelengths will undoubtedly lead to new procedures that can be performed with them. One [figure: see text] elusive use is hard tissue surgery. Although the Er:YAG has been approved for hard tissue use in the United States and currently is being used in general dentistry, it is still not yet practical or proven for large-volume osseous or extraction surgery, in which the greatest opportunity for innovation and clinical use exists. With future research, it is possible that the right wavelength laser will be developed for this purpose, allowing an increased base of procedures performed with lasers in OMS. PMID- 11048275 TI - The role of lasers in cosmetic dentistry. AB - Dental lasers contribute significantly to the field of cosmetic dentistry, providing an invaluable resource for clinicians who perform different types of esthetic procedures. Practitioners in this specialized field not only help patients acquire beautiful and ideal smiles and dental health, but also they assist patients in benefiting from tremendous clinical advantages, such as sterile surgical sites and increased comfort levels. Tooth whitening will always be an important component of cosmetic dentistry. Developing the most efficient and safe method of tooth whitening is the goal of power bleaching. Currently the argon laser has proved to be the most valuable energy source for power bleaching. Clinicians need to learn more about constantly updated technology and apply newly discovered methods and protocols to clinical situations to benefit patients and clinicians. PMID- 11048277 TI - Lasers in a hospital-based dental practice. AB - Lasers have been shown to be excellent additions to the dental armamentarium. When used on medically compromised patients in a hospital-based setting or private practice setting sufficiently equipped to deal with potential medical emergencies, lasers can make treatment of these patients much simpler and safer. As more uses are found for dental lasers and more wavelengths are developed for dentistry, lasers will become an increasingly useful tool for the hospital dental practice. PMID- 11048278 TI - Lasers in endodontics. AB - The first application of lasers in dentistry was the surgical treatment of tumors in the oral cavity, and various lasers, including semiconductor diode, carbon dioxide, helium-neon, neodymium:yttrium-aluminum-garnet, argon, erbium, and erbium chromium, have been developed since and applied in dentistry. This article describes the current and possible future clinical indications, laser devices, techniques, assessment methods, and mechanics. PMID- 11048279 TI - Practice management. AB - Laser treatment is now state-of-the-art in many disciplines of dentistry and is rapidly being recognized as the standard of care for many procedures. Seamless, efficient, and successful integration of lasers into a dental practice takes time and effort on the part of the dentist. Restructuring of fee schedules to reflect superior laser-based care and an internal and external marketing plan are essential for the dentist to recoup his or her investment in this new technology. Training the staff in laser technology and becoming familiar with new reimbursement codes (for medical and dental insurance) may become necessary. The mission statement and practice philosophy of providing superior care with less discomfort now can be realized with the integration of lasers into a dental practice. PMID- 11048281 TI - Hard tissue laser procedures. AB - A more conservative, less invasive treatment of the carious lesion has intrigued researchers and clinicians for decades. With over 170 million restorations placed worldwide each year, many of which could be treated using a laser, there exists an increasing need for understanding hard tissue laser procedures. An historical review of past scientific and clinical hard research, biophysics, and histology are discussed. A complete review of present applications and procedures along with their capabilities and limitations will give the clinician a better understanding. Clinical case studies, along with guidelines for tooth preparation and hard tissue laser applications and technological advances for diagnosis and treatment will give the clinician a look into the future. PMID- 11048280 TI - Laser curing of dental materials. AB - Research supports the use of the argon laser in dentistry. Used at powers of 250 mW +/- 50 mW for 10 seconds per increment, the argon laser provides good curing of light-activated restorative materials in a shorter period of time with equal or better physical properties as compared to the conventional halogen curing light. When used at approximately 1.5 W, it is a good soft tissue surgical instrument that cuts with little or no bleeding and minimal postoperative pain. The future looks bright for the use of the argon laser in other areas, such as decay prevention or pulpal treatments for primary teeth as well as an adjunct to endodontic therapy. PMID- 11048282 TI - Caries detection and prevention with laser energy. AB - Laser light can be used in the visible region (blue or red) as a tool for the detection of carious lesions. Techniques developed to date for early caries detection by laser light rely on fluorescence naturally from the tooth material or from bacterial by-products. Fluorescence techniques have been introduced clinically in Europe and show great promise for improved management of dental caries. Laboratory studies have shown that specific laser irradiation that is absorbed strongly by the carbonated hydroxyapatite mineral of the teeth can heat a thin layer at the surface briefly, altering its composition and making it strongly resistant to subsequent acid attack in the caries process. This resistance leads to major inhibition of subsequent subsurface caries progression and shows great promise for the treatment of susceptible sites on the tooth. This technology, if used in conjunction with ablation of carious lesions by specific laser irradiation, could prevent secondary progression around restorations. PMID- 11048283 TI - The future of lasers in dentistry. AB - In 1998, a laser-related trade journal printed an article on laser dermatology indicating that because of new applications, reduced prices, and instruments becoming more user-friendly, the dermatology laser market was becoming a billion dollar industry. If the same holds true in dentistry--keeping in mind that there are only approximately 7000 dermatologists in the United States--think of the potential for the dental laser market. It is huge, but to reach that potential, several criteria must be met: Market penetration must double in the next 4 to 5 years. Instrument sizes must diminish. Laser prices must decrease to an average of $10,000 for soft tissue lasers and $25,000 for hard tissue lasers over the next 10 years. Meeting these criteria would generate the necessary revenues for increased expenditures for research and development to improve existing delivery systems and develop new fiber types, continue development of a short-pulsed hard tissue laser to replace air turbines, and combine wavelengths into a single package, while looking into new wavelengths. If all of the above-mentioned become a reality in 10 to 15 years, the growth of the dental laser market could be limitless because of the hugeness of the worldwide dental marketplace. The last 20 years have witnessed many new developments in dental technologies, and the next 20 years promise to be even richer in technologic advancements. Lasers will be in the forefront of that growth. PMID- 11048284 TI - [Psychosocial aspects of beta-thalassemia: distress, coping and adherence]. AB - BACKGROUND: Longtime outcome in case of thalassemia depends on the patients' adherence in home treatment to reduce hemosiderosis. This study describes the patients' perspective, their typical coping strategies, health related locus-of control-beliefs and psychosocial influences on adherence. PATIENTS AND METHODS: A battery of questionnaires was employed to 43 patients with thalassemia major (3 to 26 years old) treated in Germany according to the german multicenter study respectively their parents: the Ulm Thalassemia Inventory, the KIDCOPE, the Multidimensional Health Locus of Control Scales and the Giessen Complaint List. Clinical symptoms of hemosiderosis were correlated with psychosocial variables. RESULTS: The patients feel more distressed from their treatment than from their illness itself. They react to disease-related distress with a variety of coping strategies. Some of the most frequent coping strategies are maladaptive, indicating feelings of helplessness. Internal locus-of-control-beliefs were low and fatalistic locus-of-control-beliefs were high compared with other clinical groups. The self-reported adherence to the iron chelation treatment is correlated with age, gender, age at the start-point of the treatment and emotional distress. Complaints, coping strategies and locus of control are independent from adherence as well as from hemosiderosis. CONCLUSION: Patients with thalassemia major need more information about their disease and about the benefits of iron chelation therapy. Additional psychosocial support should reduce emotional distress, strengthen coping competence and lead to a better integration of therapy in daily life. PMID- 11048285 TI - [Processes of body perception and their therapeutic use in pediatrics. From nonspecific relaxation therapy to training to recognize disease-specific symptoms]. AB - Focussing on processes of body perception is a major pathway of relaxation therapies (progressive relaxation, autogenic training, guided imagery, hypnotherapy, biofeedback). Traditionally its application has been related to psychosomatic and psychotherapeutic indications. Beyond this classical approach, recent behavioral medicine has emphasized the relevance of interoception processes and adequate attribution patterns concerning bodily sensations as a major source of adequate coping and self-management with somatic illness. Clinical application may refer to an improved cognitive-behavioral pain management in disease and treatment related conditions. Especially children and adolescents suffering from chronic conditions that may exacerbate rapidly may benefit from an education approach that teaches them to perceive their disease related complaints and symptoms accurately and to attribute them correctly. A precise, panic-free and immediate symptom recognition of sudden airway obstruction is an important precondition of adequate coping with acute asthma crisis and starting risk orientated antiasthmatic treatment. In a similar way, the child with diabetes mellitus may identify early signs of hypoglycemia by self observation, recognition and discrimination of physical, vegetative and psychological indicators of blood glucose decline that enable the child to take appropriate countermeasures. Other childhood disorders that offer chances for symptomatic self-monitoring and self-control comprise atopic dermatitis or epileptic seizures. Training young patients in precise symptom recognition may not only empower them in handling acute crisis but also strengthen global development of autonomy, control beliefs, self-responsibility and self-esteem. PMID- 11048286 TI - [Leishmaniasis with cutaneous and visceral involvement in a 13-month old boy]. AB - Leishmaniasis is an anthropozoonosis caused by infection with leishmania parasites with either cutaneous, mucosal or visceral (kala-azar) involvement. While the benign cutaneous form is self-limited death occurs in approximately 80% of children with kala-azar when untreated. The diagnosis of kala-azar should not be missed in children presenting with fever, hepatosplenomegaly and pancytopenia especially with a history of sand fly bites. We report the case of a 13-month-old boy with both cutaneous and visceral involvement. PMID- 11048287 TI - [Repeated suicide attempts by children and adolescents--results of a retrospective study]. AB - Children and adolescents who had attempted suicide once are at higher risk for further suicide attempts. In order to better identify those likely to make a further suicide attempt, they were compared at the time of their first suicide attempt with those who had only attempted suicide once. The details of all children and adolescents who had attempted suicide and been treated as inpatients or outpatients in the University Hospital in Gottingen, Germany, over a 10-year period were collected by file-analysis. Sociodemographic characteristics, stresses that preceded the suicide attempts and the chosen methods of attempted suicide were ascertained. 55 male und 116 female subjects had attempted suicide once, 17 male and 79 female subjects had attempted suicide more than once. Those with repeated suicide attempts were younger at the time of their first suicide attempt at 14 years and eight months, than those who did not make a repeat attempt, at 15 years and 7 months. Male repeaters more often experienced conflicts with their parents. Female repeaters were suffering more from emotional or chronic physical diseases and more often drank alcohol in connection with their first suicide attempt. Paediatricians, who are often the first to be confronted with suicide attempts by young people, should work towards providing psychotherapeutic support especially for children and younger adolescents with the particular stresses described. PMID- 11048288 TI - Pacifier use and sleep position in 2 to 4 month old infants. AB - Much interest has been raised in optimal infant care practices which can be preventive against sudden infant death syndrome (SIDS). In particular, avoiding prone sleep and the use of a pacifier were reported to be associated with a reduced risk for SIDS. It was suggested that potential beneficial effect resulted from the use of a pacifier might be ascribed at least in part to the prevention of an infant to be put prone to sleep and/or to turn down prone in sleep. To explore this hypothesis, the study aimed to analyse potential interaction between the habitual sleep position and the regular use of a pacifier in 2-4-month-old infants, age known for the peak incidence of SIDS. 192 randomly selected clinically healthy infants born in St. Petersburg in 1997-1998 entered the survey. The mothers were asked to complete the questionnaires addressing infant, maternal, and demographic major characteristics as well as sleep routine and other child care practices with particular emphasis on the use of a pacifier. Of 181 infants for whom the mothers were able to define typical position the baby was put to sleep and/or found, 174 (96.1%) were usually put to sleep non-prone: 103 on the back, and 71 on the side. In 51 of 181 cases (28.2%) the babies habitually changed their position in sleep. Of 174 babies usually put to sleep non-prone, 6 (3.5%) usually turned to front. Putting baby to sleep on the side compared with on the back bore the risk to turn to front equal to 3.01 (95% CI: 0.42-34.0). There was no difference in any infant, maternal, and demographic characteristic between the groups of the babies usually put to sleep prone and non-prone. Of 192 infants, 117 (60.9%) were usually left to sleep with a pacifier, and there was no significant association between the use of a pacifier and optional prone or non-prone position the baby was usually put to sleep. Likewise, the use of a pacifier did not significantly influence infant's position when found. CONCLUSION: In clinically healthy 2-to-4-month-old babies, the use of a pacifier does not have major influence on the choice of sleep position and infant's propensity to spontaneously modify it during sleep. PMID- 11048289 TI - [Peripheral lymphadenopathy in childhood--recommendations for diagnostic evaluation]. AB - BACKGROUND: Enlargement of peripheral lymph nodes most commonly caused by a local inflammatory process is frequently seen in childhood. The aim of the present study was to analyze the most common causes of peripheral lymphadenopathy and to develop a simple algorithm for the primary diagnostic evaluation of peripheral lymph node enlargement in this age group. PATIENTS: Between April and September 1999 87 unselected children (median age: 5 1/2 years) with peripheral lymphadenopathy were referred to the Department of Pediatrics, University of Graz, for further investigation. RESULTS: EBV infection was diagnosed in 20 (23.0%) children. 19 (21.8%) patients had acute bacterial lymphadenitis. In 21 (24.1%) patients lymph node enlargement was classified as "post/parainfectious (viral)". Four patients each had toxoplasmosis and cat scratch disease. In 11 (12.6%) patients neither physical nor laboratory examinations revealed pathologic results. Among the remaining 8 children sarcoidosis and Hodgkin disease was diagnosed in one patient each. Small, soft, mobile, nontender, cervical, axillary or inguinal lymph nodes do not require further investigations. In case of enlarged, tender lymph nodes with overlying skin erythema and fever diagnostic evaluation should include complete blood count, erythrocyte sedimentation rate and/or c-reactive protein level, supplemented by appropriate antibody testing (EBV, CMV, Toxoplasma gondii, Bartonella henselae). Firm, enlarged, painless lymph nodes which are matted together and fixed to the skin or underlying tissues necessitate a more detailed diagnostic evaluation in order to exclude malignant or granulomatous diseases. CONCLUSIONS: Our study demonstrated that primary diagnostic evaluation of childhood peripheral lymphadenopathy is mainly based on clinical grounds. In most cases a small number of additionally performed laboratory tests allow to correctly identify the cause of the peripheral lymph node enlargement. PMID- 11048290 TI - [Treatment of hepatitis B virus-associated membranous glomerulonephritis with interferon alfa in a 7 year old boy]. AB - BACKGROUND: The association between Hepatitis B virus infection and membranous nephropathy has been confirmed by sources in several countries. Most commonly, the illness is seen as a nephrotic syndrome. Optimal treatment remains undefined. Antiviral therapies observed with recombinant human interferon alpha may be the best treatment option. CASE REPORT: We present a 7-year old boy with membranous glomerulonephritis and nephrotic syndrome. Twelve months after the initial hospitalization therapy was started with recombinant alpha-interferon s.c. three times weekly for six months. After the therapy the patient is stable, without proteinuria, edema or renal failure. He was seronegative for HBsAg, HBV-DNA and antibody to HBeAg. CONCLUSIONS: This case report suggests that alpha interferon is effective in the complete resolution of proteinuria in HBV membranous nephropathy. PMID- 11048291 TI - [Obesity in childhood and adolescence: recommendations of a United States of America Task Force for diagnosis and therapy]. PMID- 11048292 TI - [Mineralized dental tissues: a unique example of skeletal biodiversity derived from cephaic neural crest]. AB - Molecular and structural biodiversity characterises dental mineral tissues. Groups of matrix proteins belong specifically to each tissue; amelogenins to enamel, DSPP to dentine and CAP to cementum. A wide group of proteins is also shares with other mineralized tissues such as calcium (calbindins) and phosphate (alkaline phosphatase) handling proteins. Dental tissues organisation is also based on specific cellular programs of morpho-differentiation (polarity) and on expression patterns of proteins implicated in mineralisation. The regulation of gene expression in tooth has been analysed regarding various hormones such as vitamin D in a first step and recently transcription factors (Osf-2/Cbfa1/Aml3). Other molecular families encoded by divergent homeobox genes (Msx and Dlx) are implicated in the determinism of this gene regulation and of early development. Genetic and hormonal abnormalities of dental mineralized tissues should now be interpreted thanks to the recent availability of cellular models and of odontogenic protein promoter structure. PMID- 11048293 TI - Nucleus labeling or membrane labeling for studying the proliferation of drug treated cells? AB - Proliferation and multidrug resistance status are key predictors of therapeutic outcome in acute myeloid leukemia (AML). This study compared cell cycle analysis (nuclear labeling) with cell division analysis (membrane labeling, PKH67) for studying the proliferation of cells cultured with daunorubicin (DNR) and/or Cytarabine (Ara-C), drugs commonly used in AML treatment. PKHs are a family of lipophilic, fluorescent membrane intercalating dyes. When labeled cells divide, the resulting daughter cells receive half the label, reducing fluorescence intensity to one-half that of the parent cells. DNR has the disadvantage of overlapping the spectrum of propidium iodide (PI), which is the most commonly used marker of membrane integrity. In this study, necrosis was evaluated using TOTO-3, a marker of nucleic acids emitting fluorescence above 645 nm and which incorporates cells with disrupted membranes. Comparison of cell cycle analysis with cell division for studying proliferation revealed that PKH67 was a marker of choice for analyzing the mitotic process in cells cultured with drugs. PMID- 11048294 TI - [Control of the positioning of the vertebrate limb axes during development]. AB - Limbs are formed after a series of complex interactions between tissues derived from the embryonic layers (surface ectoderm and somatopleural mesoderm). Data at both cellular and molecular levels are numerous. The developing limb becomes thus one of the best known system in the field of developmental biology. The limb bud derives from numerous embryonic tissues (surface ectoderm, somatopleural mesoderm, neural crest cells, somites, hematopoietic cells deriving from the splanchnopleura that gives rise to the aortic buds). Multiple interactions take place between these tissues. So, the definitive shape of a structure could depend on only one of its component. For example, the muscular shape is not dependent on the origin of muscle fibers but on the origin of somatopleural cells. Superior (or anterior) and inferior (or posterior) limbs differ by their shape. Some genes expressed by only one of the limb are known but genes specifying the identity of the limbs are still totally unknown. Three axes develop during the formation of the limb buds: proximo-distal, antero-posterior and ventro-dorsal. The formation of the proximo-distal axis is due to the induction of the apical ectodermal ridge, a specialized region of the surface ectoderm in which cells are prismatic and associated by gap junctions. The family of secreted FGF and their receptors play a major role in the regulation of the development of this axis. Furthermore, some genes from Hox complexes a and d participate into the regulation of the positional identity along this axis. The antero-posterior axis is determined by the zone of polarizing activity that secretes Sonic hedgehog. This molecules fashions a gradient of concentration that differentiates the future antero posterior derivatives. At last, the ventro-dorsal axis depends on the surface ectoderm that have received first positional informations from the mesoderm. The dorsal region is specified by Wnt7a and Lmx1 whereas Engrailed 1 gene plays a role in ventral specification. PMID- 11048295 TI - [Near-field microscopy: from the isolated molecule to the living cell]. AB - Near field (or scanning probe) microscopy is a recent technology which, owing to the huge amount of publications, is becoming a reference method in molecular and cellular imaging. These microscopies consist in the scanning of the sample, line by line, with a very tiny tip and thus providing informations on its surface down to the nanometer scale. These methods gather scanning tunelling microscopy (STM), which measures a current between the tip and the specimen support, atomic force microscopy (AFM), which measures the repulsive and attractive forces of the tip in contact or very close to the specimen, and scanning near field optical microscopies (SNOM), for which a glass tip allows to catch light signals. Atomic force microscopy, which allows the observation of specimens in air or physiological conditions environments, is presently dominant in biology, in complementarity with the classical optical and electron microscopies, which by the way, have also shown considerable improvements during the last years. The complementarity of these microscopies is due to their very different basic principles, which provide them various possibilities and limits. The biological applications of STM is limited by the need of conducting samples, but the different models of SNOM, often still in development, allow to consider very interesting applications, particularly for detecting very faint and tiny fluorescence signals. Different examples will be given concerning the visualization by AFM of isolated DNA molecules, naked or associated with proteins, the observation of intact or decondensed chromosomes, as well as living cells. One of the originality of AFM is its capacity to observe objects in a wide range of enlargements, with fields from a few hundred of nanometers to several micrometers. PMID- 11048296 TI - Classical cadherins in urological cancers. AB - Decreased E-cadherin expression assessed by immunohistochemistry correlates with poor survival of bladder and prostate cancer patients. The clinical usefulness of this parameter should therefore be evaluated in a large scale prospective study. In proximal kidney tubule and in its derived tumours cadherin-6 seems to take over E-cadherin function. Impaired E-cadherin function leads to increased invasive capacity of the cells. It has been shown that defective function can result from several mechanisms: mutation of the gene, alteration of transcription, post translational modifications or changes in the interaction of E-cadherin with cytoskeleton anchoring proteins: the catenins. A major mechanism leading to decreased E-cadherin expression in tumours lies in decreased transcription of the gene. Hence, a better understanding of the regulation of E cadherin transcription might open avenues for therapy by restoring a normal expression. PMID- 11048297 TI - [The proteasome and malignant hemopathies]. AB - Proteasomes are the main non lysosomal proteolytic structures of the cells. They correspond to the major system eliminating abnormal proteins, short half-life proteins and proteins controlling the cell cycle. They are essential for the production of peptides subsequently presented by the MHC-I. They are formed by a proteolytic core (the 20S proteasome) made of 4 rings of 7 proteic subunits associated with regulatory complexes (namely the 19S complex forming the 26S proteasome). Using classical cell biology techniques (cytometry, immunofluorescence microscopy, Western blot) our group has particularly studied the proteasome expression of leukaemic cell lines (U937 and CCRF-CEM) during in vitro differentiation induced by PMA and Vitamin D plus retinoic acid. During differentiation, the level of proteasome expression and its localization vary. The various monoclonal antibodies used provided different patterns according to the different subunits. There was a general trend to a disappearance of nuclear proteasome and to a decrease in their cytoplasmic expression. In contrast, proteosomal antigens were increased on the cell membrane and in culture supernatants. We derived an ELISA test to measure plasma proteasome concentrations. Preliminary results showed differences between patients with haemopoietic malignancies or solid tumors and normal donors. Proteasome levels varied under treatment. They were correlated with LDH levels. Taken together, these results argue in favor of a role for cellular proteasomes in malignant differentiation process, and emphasize the qualitative changes in proteasome expression. Plasma proteasomes do not only reflect tumor cell mass and could play a role in addition to their proteolytic activity. They seem to be a relevant tool for diagnosis, prognosis and therapeutic monitoring. PMID- 11048298 TI - [Implication of extracellular matrix metalloproteinases in the course of chronic inflammatory airway diseases]. AB - Matrix metalloproteinases (MMPs) are major proteolytic enzymes that are involved in extracellular matrix (ECM) turn over. MMP-2 (gelatinase A) and MMP-9 (gelatinase B) cleave type IV collagen, which is an important constituent of basement membrane. These enzymes play an important role in normal tissue homeostasis, but imbalance between MMPs and their tissue inhibitors (TIMPs) is thought to be a critical factor in regulating tissue remodeling. MMP-2 is produced by fibroblasts, endothelial, and epithelial cells, while MMP-9 is mainly produced by inflammatory cells. The role of MMPs was investigated through biochemical analysis or in situ expression, in the pathogenesis of two chronic inflammatory airway diseases, asthma and nasal polyposis. Both are characterized with the accumulation of active inflammatory cells, matrix remodeling and epithelial changes. Increased levels of MMP-9 and TIMP-1 were found in asthmatic subjects and NP. In NP, MMP-9 expression was detected in epithelial, endothelial and inflammatory cells. In this setting, MMP-9 could play a crucial role in the transmigration of basement membrane components by inflammatory cells leading to inflammatory cell accumulation and maintenance of inflammation in airway. Moreover, MMP-9 may contribute to cell migration, an important mechanism involved in the repair of the respiratory epithelium. PMID- 11048299 TI - [Genes implicated in glial tumors]. AB - Because of the absence of specific marker, the histological classification of gliomas remain controversial. Identifying the genetic alterations involved in gliomas makes it possible to define specific molecular pathway of tumoral progression and to define markers of prognostic and diagnostic relevance. For example, p53 mutations are frequent in low grade astrocytoma, anaplastic astrocytoma and secondary glioblastoma suggesting that it takes place at an early stage of development of astrocytic tumors, whereas inactivation of PTEN arises mainly in glioblastomas and EGFR amplification is preferentially associated with "de novo" glioblastoma. Loss of chromosomes 1p and 19q characterizes oligodendroglial tumors. However the putative tumor suppressor genes located on 1p and 19q and specifically inactivated are not known yet. Emerging technologies, like microarrays and microdissection, will allow to refine molecular data and provide a molecular classification of gliomas mechanism involved in the repair of the respiratory epithelium. PMID- 11048300 TI - Child advocacy does work. PMID- 11048301 TI - Dental caries in HIV-infected children: a longitudinal study. AB - PURPOSE: The purpose of this descriptive longitudinal clinical study was to determine primary and permanent dentition caries status in HIV-infected children, and to compare caries status with the CD4 percentage (CD4%) and immune suppression category. MATERIALS AND METHODS: 73 children up to 9 years of age with vertical HIV transmission were evaluated for caries in the primary dentition at baseline and at 6 month intervals over a 30 month period; while 19 HIV infected children between 5 and 11 years of age had their permanent dentition evaluated for caries at baseline and at 6 month intervals over a 24 month period. Caries status was also compared with CDC CD4 percentage (> 25%, 15-24%, < 15%), and CDC immune suppression categories (immune suppression: none, moderate, severe). With primary dentition caries, comparisons were made among all children (2-9 yr-olds, N = 73), < 2 yr-olds (N = 28), 2 to 4 yr-olds (N = 20), and 5 to 9 yr-olds (N = 25), and compared with NHANES III data. Caries-free status was also determined. RESULTS: During the 30-month period, there was an almost two-fold increase in primary tooth surface caries for the 2 to 9 year-olds. Caries-free status in the primary dentition declined from 60% at baseline to 37% at the 30 month period. With 5 to 11 years-olds, DMFS and DMFT remained relatively stable, while the proportion of caries-free individuals declined from 72% at baseline to 50% at 18 months. Caries in the primary dentition was increased substantially for those in the low CDC CD4 percentage categories and CDC moderate to severe immune suppression categories. CONCLUSION: Primary dentition caries status in HIV infected children is considerably greater than that for the US pediatric population, and increases with decreasing CD4 percentage and moderate to severe immune suppression. HIV-infected children with caries-free primary dentitions are less frequent than in the US pediatric population, and caries-free status decreases with age, lower CD4 percentage and moderate to severe immune suppression. PMID- 11048302 TI - A comparison of periodontal disease in HIV-infected children and household peers: a two year report. AB - PURPOSE: The purpose of this study was to compare the incidence and progression of periodontal disease in HIV-infected children to HIV-negative household peers. This paper reports the findings after two years. METHODS: Children diagnosed as HIV-infected and their household peers were recruited from the Children's Hospital AIDS Program in Newark NJ. A periodontal examination was performed at baseline and at six-month intervals for two years. A total of 121 subjects were examined two years after baseline (68 HIV-infected and 53 controls). These children ranged in age from 2-15 years at baseline. RESULTS: Plaque assessment (PHP-M) in HIV-infected cases showed a seven-fold increase over controls for the period. However, there were no significant differences between the two groups in changes over the two years for Bleeding on Probing, Gingival Index or Pocket Depths. There was virtually no recession or pathologic mobility in either group. One-fourth of the HIV-infected group exhibited Linear Gingival Erythema at both baseline and year two. Although the number of subjects with LGE did not increase, there was an increase in the severity of LGE at year 2. CONCLUSION: This study suggests that in a medically well-controlled HIV-infected population, with the exception of the prevalence of Linear Gingival Erythema, the periodontal findings are similar to their HIV-negative household peers and to the general pediatric population. PMID- 11048303 TI - Pre-eruptive intracoronal resorption as an entity of occult caries. AB - "Occult" or "hidden" caries refers to occlusal caries which is not diagnosed clinically because the occlusal surface appears ostensibly intact, and radiographs show radiolucencies in dentin. The prevalence of occult caries has been reported to range from 2.2% to over 50% of permanent molars. In spite of its relatively high prevalence, the etiology and pathogenesis of occult caries remain unclear. The author hypothesizes that occult lesions could have resulted from processes which are pre-eruptive or post-eruptive. Pre-eruptive processes include intracoronal resorption of unerupted teeth, and the post-eruptive process is occlusal fissure caries. Although the prevalence of intracoronal resorption has been shown to be around 3-6% by subject and 0.5-2% by teeth, the percentage contribution of this process to the overall prevalence of occult caries is unclear. When affected teeth are fully erupted, it is difficult to determine if pre-eruptive resorption had been present previously. The prevalence of occult lesions does not appear to be affected by fluoride exposure. Radiographs are useful adjuncts to aid in the diagnosis of occult lesions. Bitewing radiographs are useful for detecting early occlusal fissure caries while panorex radiographs of unerupted developing teeth aid in the diagnosis of pre-eruptive intracoronal lesions. It is suggested that all unerupted, developing teeth on radiographs be examined for pre-eruptive resorptive lesions. PMID- 11048304 TI - Influence of colonization with mutans streptococci on caries risk in Japanese preschool children: 24 month survival analysis. AB - PURPOSE: This study evaluates how various microbial- and salivary-related risk factors influenced the hazard for caries development in preschool children. METHODS: The study population consisted of 131 subjects (age: 0.5 to 6.0 yrs). Oral examination, including two bacterial tests and buffering capacity test, was conducted at six month intervals over 24 months. A survival analysis was used to describe caries hazard over a 24-month follow-up period. A Cox proportional hazards regression analysis was performed to test the influence of salivary mutans streptococci (MS), aciduric bacteria, buffering capacity and age on caries development. RESULTS: Of the total subjects, 60 children (46%) were found to be caries-free at baseline. Caries hazard correlated significantly with salivary MS levels at baseline (relative risk, 1.7; P = 0.003), but not with aciduric bacteria and buffering capacity. This analysis showed that all of children with high colonization of MS at baseline had dental caries 15 months later. CONCLUSION: The results suggest that salivary MS level at baseline influenced caries hazard in preschool children. PMID- 11048305 TI - Microbial contamination of toothbrushes and their decontamination. AB - PURPOSE: The objective was to determine the level of contamination of toothbrushes by mutans streptococci using microbiological identification, to access the bacterial contamination using scanning electron microscopy (SEM) and to evaluate the efficacy of two toothbrush disinfectants. METHODS: Nineteen children used their toothbrushes once a day, for five consecutive days. The toothbrushes were then immersed into disinfectant solutions for 20 h: Group I- 0.12% chlorhexidine gluconate; Group II--1% sodium hypochlorite; Group III- sterile tap water. They were then placed into test tubes containing CaSa B, for 3 to 4 days at 37 degrees C. The number of MS cfu was counted and the toothbrushes were submitted to SEM analysis. RESULTS: There was no bacterial growth in Groups I and II; Group III showed MS growth (range, 21 to 120 cfu). Scanning electron microscopy showed biofilm formation on toothbrush bristles. CONCLUSION: Immersion in 0.12% chlorhexidine gluconate and 1% sodium hypochlorite are efficient methods for toothbrush disinfection. PMID- 11048306 TI - Dental maturity in children with dystrophic epidermolysis bullosa. AB - PURPOSE: This study was performed to compare Dental Age (DA) of children with Dystrophic Epidermolysis Bullosa recessiva (DEBr) with the DA of healthy children. METHODS: Orthopantomographs (OPG's) of children with DEBr were compared with those of healthy children. Dental maturity was estimated using Dermirjian's method. A total of 48 pairs of OPG's were compared. RESULTS: There was a considerable range of variation in the difference between the chronological age and the dental age of both groups. This varied from minus 2 years 8 months to plus 3 years for the control children and minus 3 years 1 month to plus 3 years 4 months for the DEBr children. Despite this wider range the average DA of children with DEBr was statistically significantly delayed by 2 years 3 months. CONCLUSIONS: The delay in dental developmental of children with DEBr may have an impact on the clinical management of these children. PMID- 11048307 TI - Factors affecting dentifrice use and ingestion among a sample of U.S. preschoolers. AB - PURPOSE: This study was performed to assess the actual amount of dentifrice used and ingested and factors associated with use/ingestion among 28 U.S. preschoolers aged 40 to 48 months. METHODS: Using their regular dentifrice brands/flavors and small child-sized toothbrushes (Oral -B 5), the participants or their parents placed dentifrice on toothbrushes three times to assess the quantity used and its consistency. Their brushing behaviors were observed and the amounts of dentifrice ingested were indirectly measured. Afterward, the parents and children placed a "pea-sized" amount of dentifrice on their toothbrushes. RESULTS: The participants were generally consistent in quantity applied, averaging 0.256 g (range 0.035 g 0.620 g; standard deviation 0.177 g) of dentifrice per brushing. Children, either alone or with parental assistance, placed more dentifrice than either mother or father alone (P = 0.007). The estimated mean ingested fluoride was 0.17 mg F per brushing, an average of 62% of the amount of dentifrice used (range up to 98%). Amount of ingested fluoride was positively associated (P < 0.05) with the amount of dentifrice used, and negatively associated with parental assistance in brushing. When asked to apply a pea-sized quantity, the mean quantity applied was 0.314 g (range 0.064 g-0.521 g). CONCLUSIONS: This study further supports the use of small amounts of dentifrice in young children, because they ingest substantial proportions of dentifrice. PMID- 11048308 TI - Oral health knowledge, attitudes and preventive practices of third grade school children. AB - PURPOSE: The aim of study was to investigate the oral health knowledge, attitudes and preventive practices of third grade school children in Harris County. METHODS: In this cross-sectional study, oral health knowledge, attitudes, and preventive practices of the children were investigated by means of a self administered, bilingual questionnaire. Three calibrated examiners collected data on dental caries, periodontitis, and fluorosis of 1,031 school children. RESULTS: Most children reported "fairly adequate" oral hygiene habits (58%) and oral health knowledge (48%), and "adequate" dietary patterns (59%). Children with inadequate oral health knowledge were twice as likely to have caries than children with adequate knowledge (OR = 2.05, 95% CI = 1.29, 3.28). The mean combined DMFT/dft scores of children with inadequate knowledge were significantly higher than the mean for children with adequate knowledge (t = 2.6, P = 0.009). CONCLUSIONS: Results indicate a need to improve oral health knowledge and preventive practices among the study population. Because of the cross-sectional nature of the study, it is not possible to illustrate a cause-effect relationship between oral health education and prevalence of caries. PMID- 11048309 TI - Adverse reaction to amoxicillin: a case report. AB - Penicillin is the drug that most often leads to allergic reactions and anaphylaxis. The incidence of adverse events triggered by penicillins is believed to be between 1% and 10%. Up to one-tenth of these episodes are life-threatening, with the most serious reactions occurring in patients with no history of allergy. The case of a 5 year, 3 month-old female who had a severe allergic reaction to amoxicillin prior to a dental appointment is described. The literature on penicillin hypersensitivity is reviewed and recommendations for management of an allergic reaction in the pediatric dental office are discussed. PMID- 11048310 TI - Web site setup for pediatric dentists: a framework. AB - The objective of this paper is to describe a framework for the development of a simple web site for pediatric dentists. The four steps--domain name registration, web site hosting, web site design and web site publishing--are detailed. A pediatric dental web site can be created and maintained for less than $300 per annum. PMID- 11048311 TI - Correction of ectopic eruption of permanent molars utilizing the brass wire technique. AB - Ectopic eruption of a first permanent molar is a common occurrence in the developing mixed dentition. Proper management is a challenge to the pediatric dentist and is crucial to a healthy occlusion. Non-treatment can result in early loss of the second primary molar, space loss, and impaction of second premolars. Future corrective treatment may be complicated, lengthy, and costly and include: the distalizing and uprighting of the permanent molar by use of headgear and fixed or removable appliances and subsequent long term space maintenance. The brass wire technique is a fairly simple procedure and can be used successfully in moderately and even severely impacted molar cases. This procedure is described in detail with step-by-step guidelines in this paper. PMID- 11048312 TI - Treatment alternatives for sublingual traumatic ulceration (Riga-Fede disease). AB - The term Riga-Fede disease has been used historically to describe a traumatic ulceration that occurs on the ventral surface of the tongue in neonates and infants. It is frequently associated with natal or neonatal teeth but may also occur in older infants after the eruption of primary lower incisors. Failure to diagnose and properly treat this lesion can result in dehydration and inadequate nutrient intake for the infant. Treatment should begin conservatively and should focus on eliminating the source of trauma. A case is presented in which modification of sharp tooth surfaces by the Pediatric Dentist and changes in feeding techniques by the parent were used successfully to resolve this lesion. By working together, the parent and the Pediatric Dentist can achieve positive results in a short period of time with minimal trauma to the infant. PMID- 11048313 TI - Eruption of the primary dentition in human infants: a prospective descriptive study. AB - PURPOSE: This study investigated the clinical process of the emergence phase of eruption of the primary dentition including length of time taken to erupt and the association between soft tissue changes and stages of eruption. METHODS: Twenty one children aged 6-24 months at commencement of the study were recruited from three suburban daycare centers in Melbourne, Australia. Daily oral examinations of each child were conducted for seven months. RESULTS: One hundred twenty-eight teeth were observed during eruption. Swelling very infrequently accompanied tooth eruption and in all cases was mild. Forty-nine percent of observed teeth demonstrated gingival redness during the emergence stages of eruption, but there was no significant relationship between redness and specific stages of eruption. Mean duration of eruption, from palpable enlargement of the gingival tissue to full eruption, was 2.0 months (range 0.9-4.6 months). The average rate of eruption was 0.7 mm per month. Many of the deciduous teeth appeared to demonstrate an "oscillating" pattern of eruption, (emerging and then retreating before emerging again). Timing of oscillation was not specific to stage of eruption or tooth type. This was defined as a "transitional" phase of eruption which appears to be common. CONCLUSION: The results suggest that eruption of the primary dentition is often accompanied by redness, but not swelling, of the gingival tissues. For some children, there also appears to be a "transitional" eruption phase for primary teeth. PMID- 11048314 TI - Oral lymphoepithelial cyst in a young child. AB - The oral lymphoepithelial cyst is an uncommon lesion in children with rather distinct clinical features. This case report describes the pertinent characteristics of this entity, involving the floor of the mouth in a 4 year-old boy. PMID- 11048315 TI - Promoting change in children's health care. Presented to the College of Diplomates of the American Board of Pediatric Dentistry 15th annual symposium, Nashville, Tennessee, Saturday, May 27, 2000. PMID- 11048316 TI - New changes in the certification process. PMID- 11048317 TI - American Board of Pediatric Dentistry: appeal procedure. PMID- 11048318 TI - [Limits of deinstitutionalization? General aspects and developments in Sachsen. Proceedings of a meeting. Dresden, October 1999]. PMID- 11048319 TI - [Many faces of deinstitutionalization--sociological interpretation]. AB - The article summarizes in an international perspective what kind of results psychiatric deinstitutionalization has brought so far: a profound change of size and functions of the psychiatric hospital; better services for people with less severe problems; and the failing of community services to compensate for some of the functions of the former asylums, resulting in trans-institutionalization and/or neglect for many chronic patients. Three different sociological versions to explain the background and typical outcomes of psychiatric deinstitutionalization have been brought forward so far: political economy, professional dominance and post-structuralism. They are confronted with an approach using the concept of medicalisation which offers a more comprehensive understanding of the process. PMID- 11048320 TI - [Psychiatric care research--an instrument for planning health care needs]. AB - The planning of health care is primarily an administrative task. Service research supports the health administration in this task by supplying empirical data to objectify the need for care and ways to cater for that need. In this, service research on the one hand draws on descriptive and analytical data of epidemiological studies. On the other hand, indices of social structure provide information about factors influencing the utilisation of health services. Apart from this, different types of service utilisation data are used for the analysis of needs. None of the methods mentioned is sufficient on its own. Only the integration of the different levels of analysis allows sufficiently competent statements on health care needs. PMID- 11048321 TI - [Where are the limits? From institution to deinstitutionalization exemplified in Saarland]. AB - In 1998, the 1876 founded "preussische Provinzialirrenanstalt", a large asylum in Merzig (Germany), was closed down. Instead several psychiatric departments were founded in the rural county ("Bundesland"). On the asylum's former area in Merzig a new and modern general hospital including a psychiatric department was established. The present article will show up the former psychiatric hospital's genuine working fields, treating violent, self-harming and mental ill people needing help. It will describe in a differentiating way how the new services for the different groups of mental ill persons run: Decentralized general psychiatric care in defined regional areas (without forensic patients), central treatment for the forensic patients, regional care for people with mental retardation. In completion of this network there exist regional solutions for the themes "living in a own home", "Working and occupation" and "day-structuring". In conclusion, deinstitutionalization will not have any limits, despite of a few exceptions, if the limits of institutional thinking can be overcome. PMID- 11048322 TI - [Limits of deinstitutionalization?--perspective of the specialty clinic]. AB - Deinstitutionalization is a complex process that in the past was often misunderstood solely as a run down or even closure of psychiatric hospitals. Although chronically mentally ill patients were prevented from long term hospitalisation some fundamental mistakes were repeated often: patients were simply discharged without any preparations and outpatient care was badly organised. In some cases this led to therapeutic neglect, social disintegration, homelessness, incarceration or other forms of dramatic loss of quality of life. These findings are internationally confirmed although there are remarkable differences between various health care systems. Especially when primarily determined by purely economic interests deinstitutionalization constitutes a severe danger for the whole psychiatric system of care and in particular for chronic patients. Intensive research is required to avoid severe disadvantages for the further development of psychiatry. PMID- 11048323 TI - [Housing for psychiatric patients inside and outside of hospitals]. AB - The reform of psychiatric services started in Germany in 1975 with the publication of the governmental "Enquete"-Report. In the area of housing for chronic patients the recommendations made 1975 have not been fulfilled. A greater number of chronic patients as planned live in institutionalised homes with little access to the community and shortcomings in the area of individual self determination. The process of deinstitutionalization of the old chronic patients from psychiatric hospitals has created new problems. Some 60,000 patients have been deinstitutionalized, but in some parts of the country to a considerable degree just by renaming parts of the hospital into a home for the disabled. There is a need for research on who lives where and how and efforts have to be made to change this situation by the development of more community centered and individualised forms of living as shown by many successfully working regions in the country. PMID- 11048324 TI - [Limits of deinstitutionalization--perspectives of relatives of psychiatric patients]. AB - After a hopeful beginning, the social process of the reintegration of those with severe mental illness has come to a standstill. I am led to wonder whether "the community" really wants to live together with people suffering from severe mental illness, and if so, how closely? As long as the medical treatment of mental illness provided by the general practitioners is fundamentally deficient, as they are not able to prescribe the necessary interventions--such as out-patient psychiatric nursing, and service providers in the out-patient sector are content with offering increasingly intensive forms of care for the less seriously ill at the cost of the Social Welfare System--the reintegration of those with serious mental illness remains an illusion--which is mainly to the benefit of providers of residential care in homes and hostels. PMID- 11048325 TI - [Limits of deinstitutionalization: experience in England]. AB - Psychiatric reform and the current system of mental health care in England are outlined, the system context of the National Health Service (NHS) is described. Recent institutional change in the NHS has introduced internal market elements. More than 90 mental hospitals have been closed. The TAPS study, a longitudinal study evaluating mental hospital closure is described. Results show a range of advantages of the post-discharge care arrangements. Studies evaluating services with a home treatment and community focus (DLP, PRiSM) suggest that community mental health care is feasible. Shortages and problems of the current system of care (acute beds, intensive residential and rehabilitation services) are outlined. PMID- 11048326 TI - [Problems in the treatment of mentally ill offenders--a problem of general psychiatry?]. AB - The special position of psychiatry among the various medical disciplines is determined not least by the phenomenon of violent behaviour of some of its patients and the possibility of coercive measures against patients. The worldwide process of deinstitutionalization since the last decades is characterized by a substantial reduction of inpatient treatment and the expansion and improvement of increasingly specialized community care offers. Yet, at the same time paradoxically we are confronted with an increasing neglect of the special care requirements of a certain group of difficult-to-place patients (mostly severely chronically ill with high rates of comorbidity). Despite different social and legal conditions this has uniformly led to an increasing shift of these patients into the forensic system which is illustrated by a comparison of international and Austrian data. Forensic psychiatry is hardly able to cope with this development because of structural and personal deficits existing and is in danger of being misused primarily as an instrument of social control. From the position of forensic psychiatry the limits of deinstitutionalization are set by the feeling of responsibility of general psychiatry for a subgroup of troublesome and difficult to treat patients. PMID- 11048327 TI - [Psychiatric care in Saxony]. AB - A fundamental policy discussion of the scope and purpose of care for the mentally ill is presented along with practical examples. The second part describes the development of psychiatric care in the Free State of Saxony since 1990. Finally, the author outlines future developments. PMID- 11048328 TI - [Comments on deinstitutionalization]. AB - Following a historic introduction the development of the chronics' departments of the big psychiatric hospitals in Saxony since 1990 is described: the goal of ending the care of the chronic mentally ill younger than 65 years in these departments, the newly restructured care of the mentally ill in Saxony and the problems attached to these processes. PMID- 11048329 TI - [Social welfare in the care of chronically ill psychiatric patients]. AB - The article shows the development of the long-stay departments at the psychiatric hospitals in Eastern Germany from the perspective of a local government. Against the apparently increasing scepticism of concepts of de-institutionalisation, the example of the city Leipzig is used to show how the process of reintegrating residents of long-stay institutions can be supported, and how decentralized out patient psychosocial services--which can also be used by the community inhabitants--can be built up at the same time. Reference is also made to the well developed out-patient services and clearly structured treatment offers for the inpatient sector which have already existed in the city before the political changes in Eastern Germany. Further, the role of the local Social Welfare Office in the realization of de-institutionalisation projects will be outlined. PMID- 11048330 TI - [Perspectives of psychiatric care in Leipzig--deinstitutionalization from the viewpoint of neurologist/psychiatrist in private practice and the work of consortium of community psychiatric services]. AB - Two perspectives of mental health care in Leipzig are outlined. Critical aspects of deinstitutionalization are discussed from the point of view of an office-based Nervenarzt (neurologist and psychiatrist). The limitations of office-based practice in providing care for the severely mentally ill (SMI) are described, i.e. lack of a multidisciplinary community mental health team, community psychiatric nursing and social work back-up in particular. Residential service and nursing homes are often under-staffed and ill-prepared for caring for people with SMI. A second view-point describes the Verbund Gemeindenahe Psychiatrie, a community psychiatric service for the just under 500,000 population of Leipzig in seven community mental health centers each combining day hospital, out-patient clinics and multidisciplinary community psychiatric care. This service is unique in Saxony and well accepted by service users and professionals. PMID- 11048331 TI - A review of health-based comparative risk assessments in the United States. AB - Comparing the risks posed by specific environmental hazards has become attractive to policy makers and legislative bodies as an aid to budgeting and other policy decisions. This paper reviews the human health-based findings from the first federal comparative risk assessment project and subsequent reviews conducted by 15 states and local government agencies in the United States. Methods are described on conducting comparative risk assessments that include substantive involvement of the public and special interest organizations. A consolidation of the comparative risk assessments of 15 states revealed good agreement with federal health-based environmental hazard priorities and partial agreement with local-government health departments. In descending order of priority, indoor air pollutants (excluding radon), criteria air pollutants, hazardous air pollutants, indoor radon, lead contamination, inactive hazardous waste sites, and drinking water at the tap are the highest ranked environmental hazards to human health. PMID- 11048332 TI - Anthropogenic emissions of carbon monoxide. AB - Carbon monoxide (CO) is a colorless, nonirritating, odorless and tasteless gas. Carbon monoxide combines with hemoglobin far more readily than does oxygen, leading to tissue hypoxia. Thousands of people die annually from CO poisoning, and those recovering from acute exposure commonly suffer brain damage. Chronic poisoning is of particular concern to sufferers of coronary heart disease, pregnant women, and people with certain hematological disorders. Indoor emission sources, notably fuel-burning heating appliances, cause most unintentional deaths and cases of illness and should be the main focus of concern. Motor vehicle emissions pose a chronic health risk for occupationally exposed groups. Smoking is a major source of personal exposure. Recent exposure to CO is commonly evaluated by measuring blood carboxyhemoglobin levels, which are related to the concentration of atmospheric CO. Monitoring methods are reviewed here, and monitoring is considered in relation to air quality standards and guidelines. Finally, control measures for motor vehicles and indoor heating appliances are suggested. PMID- 11048333 TI - Cadmium in the environment: sources, mechanisms of biotoxicity, and biomarkers. PMID- 11048334 TI - Effects of environmental pollutants on airways, allergic inflammation, and the immune response. AB - Particulate and gaseous air pollutants are capable of damaging the airway epithelial lining and of shifting the local immune balance, thereby facilitating the induction of persistent inflammation. Epidemiological studies are inconclusive regarding whether air pollution increases the incidence of asthma and chronic bronchitis in the population. Clearly, environmental pollution can, however, precipitate attacks and emergency-room admissions in those already suffering from such conditions. The catastrophic potential of airborne pollution was demonstrated in the 1960s and 1970s, when inverted atmospheric pressure conditions trapped smog over cities on the Eastern coast of the United States and over Europe. This smog resulted in thousands of hospital admissions and dozens of deaths. With the general rise in the incidence of atopy and asthma in the Western population, it is of major public health interest to reduce, as much as possible, the exposure of such populations to anthropogenic and natural sources of pollution. PMID- 11048335 TI - Association between malignant tumors of the thyroid gland and exposure to environmental protective and risk factors. AB - Risk factors for thyroid carcinomas and adenomas were investigated using a standard questionnaire in a case-control study in Southwestern Germany, a known iodine deficiency area. A clinical registry, set up after the Chernobyl accident at the University hospital Mannheim, served as the basis for 174 incident cases of each diagnostic group. Interview data were compared within and with prevalences from a population-based matched control group of equal size from the entire area. The protective role of coffee drinking and the consumption of cruciferous vegetables, such as broccoli, were confirmed for both genders. A high consumption of tomatoes (> 200/year) was associated with an elevated risk of > 2.5 for malignant tumors but not for benign tumors in both genders. In both genders, both treatment for goiter (hyperthyroidism) and decaffeinated coffee consumption were associated with an increased risk for malignant tumors, but less so for adenomas. In women, early menarche (< 13 years) and stillbirth after first pregnancy, as well as hysterectomy, were substantial risk factors. Occupational variables and radiation, including medical indications and mammography, did not reveal particular risks. We did not address the role of regular iodine substitution, but did analyze the consumption of freshwater fish and seafood. Multivariate analyses of the most prominent risk factors confirmed the persistence of tomato consumption as a risk factor. In view of experimental evidence on the carcinogenicity of organophosphates and the neurotoxicant effect of certain agrochemicals on neuroendocrinologically regulated organs, we postulate that in Germany, importing off-season tomatoes from areas with a known history of possible inexperienced use of agrochemicals may be associated with a promoting effect for malignant neoplasias of the thyroid gland in terms of promoting already existent proliferating tissue growth. PMID- 11048336 TI - [Palliative care--a challenge]. PMID- 11048337 TI - [Development and state of palliative treatment in Germany]. AB - Since the early nineties of the 20th century palliative medicine developed in a dynamic way. 65 palliative care units (p.c.u.), 81 hospices and some 600 outpatient services were in existence in spring 2000. Germany provides 7 beds in p.c.u. and 8 beds in hospices per 1 Mill. inhabitants. Stillitt is a long way to diminish the existing deficits in pain therapy, the control of other physical symptoms as well as the psychological, social and spiritual support. PMID- 11048338 TI - [Ambulatory palliative care--special qualities]. AB - During the last phase of progressive cancer numerous problems may arise. Additionally to pain and other symptoms, psycho-social and spiritual considerations supervene. In such cases palliative medicine offers positive options. In Germany, these options have been made available mainly for in patients (at palliative care units or hospices). But out-patient care must continue to be a priority, since most patients wish to remain at home as long as possible and die at home. There are various problems that also arise at a patient's home. As the possibilities of palliative medicine are still generally unknown in out-patient care, new structures of medical care have to be established. This is where home care services come in; staff, especially educated professionals supported by volunteers assist the patients at home, together with general practitioners and health and advice centres. The work of home care services focuses on supervising pain therapy and symptom control as well as psychosocial advice and support of patients and their families. The statistics of the home care service Bonn shows that in 1999 73 per cent of patients could be treated at home until the end of live. In 43 per cent physicians made the initial contact with this service organisation. This proves the necessity and acceptance of new structures in palliative out-patient care. PMID- 11048339 TI - [Basics of drug therapy of cancer pain]. AB - Industrial countries experience a significant increase of cancer prevalence. Despite recent advances in the treatment of various types of cancer still most of the patients cannot be cured. Especially the advanced incurable stages of cancer, however, often are accompanied by severe pain. Therefore, the high demand for a sufficient pain management and symptom control seems obvious. Throughout the last decades new drugs and techniques for the management of cancer pain have been developed. Most cancer patients should experience sufficient pain-management if existing recommendations for the pharmacological treatment of cancer pain (e.g. WHO-guidelines) are followed consequently. If, nevertheless, intractable pain or ongoing disabling symptoms continue despite proper therapy, every doctor should feel himself obliged to consult an expert in palliative medicine, in order not to tolerate avoidable suffering of his patient. PMID- 11048340 TI - [Treatment of constipation and different laxative requirements in palliative medicine]. AB - The significance of constipation with its variety of possible complications is often underestimated in the context of the tumour patient's complaints although difficulties in stool management are more common in patients with advanced cancer than in those with other terminal diseases. Without treatment constipated patients will suffer from nausea and emesesis and will possibly develop small bowel paralysis. About half of all patients admitted to specialist palliative care units report constipation, but about 75% of the patients will require laxatives. Unlike for pain, no generally accepted and widely disseminated management guidelines are available. Effective prophylaxis and cause-based therapy do improve the alimentary condition and can help to prevent the transition to ileus situations. Effective symptom management presupposes exact knowledge of the pharmacokinetics. PMID- 11048341 TI - [Treatment of nausea nad vomiting in palliative medicine]. AB - Nausea and vomiting are the most distressing gastrointestinal symptoms in patients with far advanced malignant diseases. A complex pathophysiology exists between gastrointestinal tract and brainstem. Neurotransmitters play an important role. An exact history and physical examination are necessary for the choice of the antiemetic. Dependent on the cause the antiemetic, which blocks receptors peripherally or centrally, will be chosen. Main antiemetic groups are prokinetics, 5HT3-antagonists, dopaminantagonists, antihistaminics and phenothiazines. Symptom relief can be reached in 90% of the patients with a differentiated approach. PMID- 11048342 TI - [Nutritional problems in palliative medicine]. AB - Malnutrition is a frequent problem in the palliative care of the seriously ill and dying. Want of appetite and los of weight are direct symptoms of patients with consumptive infectional diseases (AIDS, TBC) as well as cancer or geriatric patients. Severe malnutrition significantly contributes to a loss of quality of life and increases morbidity of palliative patients. The subjective well-being of seriously ill patients is heavily influenced by want of appetite and loss of weight. Patients often find want of appetite and the incapability to eat as pressing as the physical impairment caused by the disease. Therefore the sole aim of palliative dietotherapy has to be to strengthen the general physical and mental condition of the patient. A specific training of home care staff and relatives of seriously ill patients in dealing sensitively with this problem of care is desirable. Above all, in-patient treatment of affected patients for the sole purpose of feeding has to be avoided. Aggressive dietotherapeutic interventions, especially artificial feeding, should be refrained from as far as possible in the terminal phase. Only if the prognosis of a patient in palliative treatment is improving contrary to expectations are strategies of curative dietotherapy valid. PMID- 11048343 TI - [Consideration, assessment and treatment of difficulty in breathing in palliative medicine]. AB - Next to pain, the most frequent fear of dying patients, their relatives and doctors in attendance is to suffocate, even if there is no significant dyspnoea. During their progress of disease, around half (40-60%) of all tumour patients suffer from difficulty in breathing. This is due to many reasons, which include the entire differential diagnosis of dyspnoea as well as psychological and social aspects. Lacking any useful therapeutic consequence, there is no reason for extended diagnosis in palliative medicine. Treatment of symptoms should not be delayed by diagnostic measures. Besides application of demand drugs as morphine and lorazepam the most important therapy against asphyxia is the individual treatment of symptoms. All people involved should be educated in general treatment, nursing and psychological care to reduce the dying patient's fear of suffocation. This will result in the reduction of the patients states of panic and therefore allow them to die in a more peaceful way. PMID- 11048344 TI - [Quality assurance in palliative medicine. Survey of the structure and processing quality in palliative care units in North Rhine-Westphalia in Germany]. AB - BACKGROUND AND METHODS: The aim of this study was to get detailed information about the current situation and the quality of the palliative care units in North Rhine-Westphalia (NRW). The aim of palliative medicine is the achievement of the best possible quality of life for patients and their families. Unrelieved pain and other symptoms or major social problems are the reason for the admission of a patient to a palliative care unit. Questionnaires were distributed to the 13 palliative care units in NRW. RESULTS: Most palliative care units in NRW focus on the achievement of pain relief and symptom control, trying to achieve the best possible quality of life. However, the quality of palliative care shows some significant deficits (in the availability of nursing staff, cooperation with general practitioners, standardised documentation and education). A multi professional team is available in only four units. A total of 90 beds were available in NRW. In 1998 and 1999 palliative care units cared for 2308 patients, most of them (97.5%) suffering from cancer. CONCLUSION: There is a need for further education, not only for physicians but also for nursing staff and physicians already working in palliative care units. Furthermore, we need specialists in palliative medicine for the care for patients with particularly severe problems, and to initiate educational programmes and research in palliative medicine. In order to achieve an improvement of palliative care in Germany, we need to convince not only physicians and nursing staff of the advantages of palliative care, but also health care officials, the government and the public. Palliative care is not for free. However, palliative care does not necessarily lead to increasing costs in health care. Better pain management and symptom control may help to save the overall costs of medical treatment. PMID- 11048345 TI - [Psychosocial aspects in terminally ill and dying patients]. AB - Considering the experiences and behaviour patterns of terminally ill patients, especially their perceptions, fears, wishes and needs, the classical phases as possible stages in the processes involved in the last big crisis in life are newly described. Consequences on how to adequately communicate and interact with these patients are represented. Psychological and social support of the dying person and their relatives demands preparation as well as accompanying (e.g. supervision and education) of an interdisciplinary team (in the context of a hospice or palliative treatment). Psycho-social offerings and interventions are discussed as well as subsequent support of the bereaved. Delimitation of psycho social, psychological and religious support competency is outlined. The article ends with the questions which remain to be researched and conclusions. PMID- 11048346 TI - Possibilities and limitations in priority setting in German ambulatory health care. AB - Ambulatory health services provided by physicians in Germany get reimbursed by the statutory health insurance when benefits and costs of a proposed technology compared to alternatives are favourable. This decision has to be made by a joint committee of physicians and sickness funds which has far-reaching autonomy in health issues (the so-called "Bundesausschuss"). In order to offer a comprehensive basket of health benefits it is necessary to develop an explicit, transparent, and reproducible prioritization process which allows the evaluation of new technologies as well as the evaluation of already introduced technologies. The existing approach tends to be uncomprehensive and neglects the potentials of steering health care towards defined health goals. However, it reflects a bottom up procedure which meets with physicians' as well as patients' needs. For existing technologies a framework specific for Germany is proposed which allows the evaluation of health technologies in a context with health goals. Already implemented technologies should be evaluated in a conceptual framework which allows the implementation of mid- and long term health goals for German ambulatory health care. To achieve this goal a "procedural" consent of participating stakeholders is necessary. PMID- 11048347 TI - [From data entry to data presentation at a clinical workstation--experiences with Anesthesia Information Management Systems (AIMS)]. AB - Anesthesia Information Management Systems (AIMS) are required to supply large amounts of data for various purposes such as performance recording, quality assurance, training, operating room management and research. It was our objective to establish an AIMS that enables every member of the department to independently access queries at his/her work station and at the same time allows the presentation of data in a suitable manner in order to increase the transfer of different information to the clinical workstation. Apple Macintosh Clients (Apple Computer, Inc. Cupertino, California) and the file- and database servers were installed into the already partially existing hospital network. The most important components installed on each computer are the anesthesia documenting software NarkoData (ProLogic GmbH, Erkrath), HIS client software and a HTML browser. More than 250 queries for easy evaluation were formulated with the software Voyant (Brossco Systems, Espoo, Finland). Together with the documentation they are the evaluation module of the AIMS. Today, more than 20,000 anesthesia procedures are recorded each year at 112 decentralised workstations with the AIMS. In 1998, 90.8% of the 20,383 performed anesthetic procedures were recorded online and 9.2% entered postopeatively into the system. With a corresponding user access it is possible to receive all available patient data at each single anesthesiological workstation via HIS (diagnoses, laboratory results) anytime. The available information includes previous anesthesia records, statistics and all data available from the hospitals intranet. This additional information is of great advantage in comparison to previous working conditions. The implementation of an AIMS allowed to greatly enhance the quota but also the quality of documentation and an increased flow of information at the anesthesia workstation. The circuit between data entry and the presentation and evaluation of data, statistics and results directly available at the clinical workstation was put into practice. PMID- 11048348 TI - [Treatment guidelines--cancer pain. Recommendations for therapy of cancer pain from the Medical Commission of the German Medical Society, Second Edition, April 2000] . PMID- 11048349 TI - Addicts' choice. PMID- 11048350 TI - Selling drink to underage adolescents and getting away with it. PMID- 11048351 TI - Shakespeare and the meaning of authorship. PMID- 11048352 TI - Opium use in Turkmenistan: a historical perspective. AB - This paper presents a picture of how the patterns of opium use have changed in Turkmenistan over more than 100 years and the relationship between these transformations and formal and informal social controls of drug use. From the late 18th century, when opium use began to become a social problem, informal control weakened. Eventually, in the late 19th century, formal control was introduced, aimed at the prohibition of drug trade and use. From that time, the intended and unintended outcomes of implemented policies led to changes in the demographic patterns of users and the social-medical consequences of opiate use. The anti-drug policies, where criminal prohibition coexisted with strategies aimed at raising the population's general living standards and at providing free access to health care, were effective up to the early 1980s. New political and social-economic realities in the 1980-90s have radically changed the drug scene in the country, with heroin trade and use as the main concerns. The government's reaction, while following the old paths, has included new elements, based mainly on ideas of national consolidation. PMID- 11048353 TI - One-year follow-up of disulfiram and psychotherapy for cocaine-alcohol users: sustained effects of treatment. AB - AIM: To evaluate outcomes 1 year after cessation of treatment for cocaine- and alcohol-dependent individuals. DESIGN: Randomized controlled trial. SETTING: Urban substance abuse treatment center. PARTICIPANTS: Ninety-six of 122 subjects randomized to treatment. INTERVENTIONS: One of five treatments delivered over 12 weeks. Cognitive-behavioral treatment (CBT) plus disulfiram; Twelve-Step facilitation (TSF) plus disulfiram; clinical management (CM) plus disulfiram; CBT without disulfiram; TSF without disulfiram. MEASUREMENTS: Percentage of days of cocaine and alcohol use during follow-up, verified by urine toxicology screens and breathalyzer tests. RESULTS: First, as a group, participants reported significant decreases in frequency of cocaine, but not alcohol, use after the end of treatment. Secondly, the main effects of disulfiram on cocaine and alcohol use were sustained during follow-up. Finally, initiation of abstinence for even brief periods of time within treatment was associated with significantly better outcome during follow-up. CONCLUSIONS: These findings support the efficacy of disulfiram with this challenging population and suggest that comparatively brief treatments that facilitate the initiation of abstinence may have long-term benefits. PMID- 11048354 TI - Measuring injecting risk behaviour in the second decade of harm reduction: a survey of injecting drug users in England. AB - AIMS: To measure risk behaviour among injecting drug users (IDUs) using the Injecting Risk Questionnaire (IRQ). METHODS: Data were analysed from the first multi-site survey of injecting risk behaviour among IDUs not in contact with drug services in England. A total of 1214 IDUs were recruited from community settings in seven sites. FINDINGS: Fifty-two per cent reported sharing injecting equipment in the previous 4 weeks in response to a single question on sharing. This rose to 78% when asked more detailed and multiple questions on injecting risk practices. Levels of injecting risk behaviour did not differ substantially by gender, age, length of injecting career, main drug of injection, previous treatment contact or geographical location. However, sharing partners were restricted to a median of two others. CONCLUSION: These data raise questions concerning the extent to which levels of injecting risk behaviour have increased over recent years, or the extent to which previous monitoring systems underestimated levels of risk. None the less, the data confirm that the promotion of safer injecting continues to be an important public health issue with regard to reducing blood-borne infections. PMID- 11048355 TI - A longitudinal study of male buprenorphine addicts attending an addiction clinic in India. AB - AIM: There is a lack of longitudinal studies of buprenorphine dependence, an important opioid dependence in several countries. We investigated the course and outcome of buprenorphine dependence in an Indian clinic-attending cohort. DESIGN: Retrospective longitudinal study. SETTING: An addiction clinic in northern India. PARTICIPANTS: Ninety-four male patients with buprenorphine dependence, registered for treatment between 1987 and 1993. Follow-up analyses were conducted for the 52 patients (55% of the index cohort) who completed more than a year of follow-up. In 48% of these 52 patients data were obtained from their clinical records of follow-up, while 52% were contacted specifically to obtain the required data on follow up. MEASUREMENT: Baseline demographic and clinical variables; time spent in various phases of use or abstinence; outcome at the latest follow up; transition to other drugs during follow-up period. FINDINGS: Over an average follow-up duration of 3 years, 56% of the time was spent in dependent use, 12% in non-dependent use and 32% in abstinence. By the end of follow-up, 6% of patients were dead (annual death rate 1.9%), 33% were unchanged and 61% were classified as "improved". The proportion of patients with "improved" outcome increased over the years. Patients with poor outcome had shorter follow-up and hospital stay, and had used pentazocine and/or antihistaminic injections in the buprenorphine "cocktail" more often than those with better outcome. Thirty-two patients shifted to other drugs over the years, notably heroin or polydrug use. These "transition" patients had a family history of drug use more often, started their drug career earlier, had marital and legal complications more often, spent more time in dependent phase of drug use, underwent multiple hospital admissions but stayed for a shorter period and faced more deaths, when compared to those who did not shift. CONCLUSION: In clinic-attending male patients with buprenorphine dependence who were followed-up although dependent pattern of use of the drug continued for a long time in their career, there was a slow but progressive improvement. Transition to other drugs was associated with a worse course and outcome as compared to being stable on buprenorphine. PMID- 11048356 TI - Alcohol sales to underage adolescents: an unobtrusive observational field study and evaluation of a police intervention. AB - AIMS: The aims of this study were to assess the ease with which adolescents in the United Kingdom are able to buy alcohol, to obtain information concerning vendors' perceptions of alcohol sales to adolescents, and to evaluate a police intervention intended to reduce underage alcohol sales. DESIGN, SETTING, SUBJECTS: An unobtrusive naturalistic field study was conducted in two urban locations. Pairs of 13- and 16-year-old boys and girls were trained to attempt the purchase of different types of alcohol (alcopops, beer, cider, wine, spirits) from four different types of retail outlets (corner shops, off-licence, public houses and supermarkets), under the supervision of a researcher and typically a parent. The assessment was repeated, with the omission of the 13-year-old boys, following a police intervention in one of the performance sites, consisting of warning letters and visits to vendors, and the issue of a small number of police cautions. A total of 62 underage confederates in all attempted 470 test purchases in phase 1 and 348 in phase 2. Between the two waves of test purchases a sample (n = 95) of the same vendors was surveyed by telephone. FINDINGS: In phase 1, sales resulted from 88.1% of purchase attempts by 16-year-old girls, 77% of attempts by 16-year-old boys, 41.6% of 13-year-old girls and 4.1% of 13-year-old boys. These figures were generally comparable across locations, alcohol types and outlet types. Refusals were more likely when another vendor was present. Eighty per cent of sales to 16-year-olds and 65% of sales to 13-year-old girls were made without challenge. "Prove-It" ID cards were requested in fewer than 12% of purchase attempts in both age groups. Overall, there was no evidence that the police intervention reduced sales of alcohol to 16-year-olds. There was a hint that the intervention may have caused a very short-lasting decrease in sales to 13-year-old girls, but this was contained within an overall increase in sales to this group. Alcohol vendors reported that they rarely encountered underage customers or refused sale though 90% of vendors said that if they became suspicious, they would request ID. Only two vendors believed that they were likely to suffer adverse consequences if they sold alcohol to minors. CONCLUSIONS: These data suggest that 16-year-olds, and girls as young as 13, have little difficulty in purchasing alcohol, and that there is little difference between different types of outlets in their willingness to sell alcohol to minors. Vendors perceive little risk in selling alcohol to adolescents. The fact that the police intervention failed to decrease sales suggests that vendors do not change their behaviour in response to the threat of legal action. PMID- 11048357 TI - Prevalence of alcohol use and the association between onset of use and alcohol related problems in a general population sample in Germany. AB - AIMS: This paper examines the prevalence of alcohol use, alcohol-related problems and onset of regular alcohol use, including its association with the prevalence of CAGE symptoms. DESIGN AND PARTICIPANTS: Data come from three nationally representative samples of the non-institutionalized German general population aged 18-59 years which were conducted annually between 1994 and 1996. Subjects (n = 7501) were surveyed through telephone interviews. FINDINGS: Overall, men were more likely to drink alcohol, to be heavier drinkers and to experience more alcohol problems than women. Prevalence of 12-month use was constant across age among males, but decreased with age among women. Prevalence of heavier drinking, however, increased with age in both sexes. While for both sexes the median age of onset decreased towards younger cohorts, the prevalence of regular use at younger ages increased more strongly among females compared to males. In all cohorts of both sexes, an association between early age of onset and negative consequences measured by the CAGE questionnaire could be observed. Odds ratios were significantly lower for life-time experience of at least two CAGE symptoms among respondents if they had started regular alcohol use later than their peers. CONCLUSIONS: The results indicate a narrowing of the gender gap due to an increasing prevalence of regular alcohol use for females across cohorts. Gender differences with regard to heavier drinking are still prominent. The findings confirm those of related studies in which early age of onset of regular alcohol use was found to be a significant predictor for life-time alcohol-related problems. PMID- 11048358 TI - Relationship between follow-up rates and treatment outcomes in substance abuse research: more is better but when is "enough" enough? AB - AIMS: To examine the effects of different follow-up rates on estimates of treatment outcome and predictive models thereof, and to specify participant characteristics associated with tracking difficulty. DESIGN: An observational study using data collected for a randomized, experimental design. SETTING: The King County Assessment Center in Seattle, Washington, an organization responsible for referral to publicly funded substance abuse treatment. PARTICIPANTS: Substance-addicted individuals referred to publicly funded inpatient or outpatient treatment. MEASUREMENTS: Standardized self-report instruments measuring substance use, substance use consequences and general functioning. Chart review was used to measure treatment entry and completion. FINDINGS: There was a significant association between follow-up difficulty and outcomes related to addiction treatment and later substance use. However, outcome estimates based on 60% of the sample who were easiest to locate were only minimally different from those based on the 90-100% ultimately captured, and predictive models of outcome based on the 60% group were reasonably similar to those based on the final sample. Of baseline characteristics examined, only age was associated with later tracking difficulty. CONCLUSIONS: Studies reporting follow-up rates below 70% may produce valid findings and study attrition may be largely unpredictable from participant characteristics at baseline. However, a number of factors such as type of population studied, geographical location of the sample, reasons for loss to follow-up and sample size must be considered when attempting to generalize the findings of this study. PMID- 11048359 TI - Maternal addiction, child maladjustment and socio-demographic risks: implications for parenting behaviors. AB - AIMS: In this study we examined three parenting dimensions (involvement, autonomy, and limit-setting) and three potential determinants (maternal addiction, low SES and its correlates, and mothers' perceptions of their children's maladjustment) in order to disentangle features of parenting that are uniquely related to maternal addiction from those related to contextual determinants. We also examined conditional effects of low SES and its correlates on parenting. DESIGN: Based on a literature review and predictions arising from an ecological model of parenting, we expected that maternal addiction would be related with problems in parental involvement, but that the other parenting dimensions would be related with mothers' perceptions of children's maladjustment and low SES. Accordingly, we examined variance in each parenting dimensions accounted for by each of the three determinants, respectively. PARTICIPANTS: Subjects included 120 (69 opiate-addicted and 51 SES-matched comparison) mothers with children under 16 years of age. MEASUREMENTS: Children's maladaptive behavior was assessed with the Behavioral Assessment System for Children, and parental adjustment with the Parent Child Relationship Inventory. FINDINGS: Direct effect predictions were confirmed and two conditional effects involving single status and family size were also found. CONCLUSIONS: Although many parenting problems have previously been attributed to maternal addiction, only parental involvement is directly related to being an addict; other parenting dimensions may be better explained by contextual factors. PMID- 11048360 TI - "Perils of partnerships": comments on an editorial. PMID- 11048361 TI - Drinks industry response. PMID- 11048362 TI - Comment on ICAP. PMID- 11048363 TI - Responsible drinking in Russia: whose dream? PMID- 11048364 TI - Psychological problems of partnership and profit or: who has the higher population impact? PMID- 11048365 TI - ICAP and the new terminology. PMID- 11048366 TI - The Stroop colour-naming task and addictive behaviour: some recommendations. PMID- 11048367 TI - The dangers of self-examination. PMID- 11048368 TI - Mild whiplash lasting more than a month. PMID- 11048369 TI - Key developments in men's health. PMID- 11048370 TI - Casebook: PSA dilemmas. PMID- 11048371 TI - A rational approach to prostate cancer. PMID- 11048372 TI - A guide to common conditions of the penis. PMID- 11048373 TI - The GP's role in HIV and AIDS care. PMID- 11048374 TI - The patient with haemospermia. PMID- 11048375 TI - Gynaecomastia: when is action required? PMID- 11048376 TI - The growing problem of antibiotic resistance. PMID- 11048378 TI - The MRCGP. A blueprint for success. PMID- 11048379 TI - Diagnosis treatment. PMID- 11048377 TI - Diagnosis and management of Addison's disease. PMID- 11048380 TI - Piercing difficulties. PMID- 11048381 TI - Primary pulpotomies. PMID- 11048382 TI - Blood donors ban. PMID- 11048383 TI - Changing procedures? PMID- 11048384 TI - Mobile telephones and lesions of the mouth. PMID- 11048385 TI - Antibiotic prescribing for sinusitis? PMID- 11048386 TI - NHS discrimination? PMID- 11048387 TI - Success against oral cancer? PMID- 11048388 TI - Tobacco addiction. PMID- 11048389 TI - Slipshod approach? PMID- 11048390 TI - Temazepam dangers. PMID- 11048391 TI - Educating dentists--the value of the research environment. AB - I hope, ladies and gentlemen, that you agree with my opening statement that 'during the immediate past century, a dramatic improvement of oral health and dental status occurred on a world wide basis.' In my opinion, the combined efforts from dental educators, dental researchers and dental clinicians have, without doubt, been the actual initiators and generators for this improvement working along the traditions of dental scientists such as Sir Wilfred Fish. PMID- 11048393 TI - Tooth preparation techniques for porcelain laminate veneers. AB - OBJECTIVE: The purpose of this study was to determine the effect that two guides to tooth preparation had on an operator's ability to appropriately and consistently prepare teeth for porcelain laminate veneers. STUDY DESIGN: In-vitro study. MATERIALS AND METHODS: Thirty typodont central incisor teeth were randomly allocated into three groups and a general dental practitioner was asked to prepare the teeth for porcelain laminate veneers. Group A were prepared freehand while Groups B and C were prepared with the assistance of a silicone index and depth preparation bur respectively. Images of the prepared teeth were used to calculate the mean labial depth of preparation and incisal reduction of teeth in each group. RESULTS: The mean labial reduction for Groups A, B and C was 0.37 mm (SD 0.13), 0.62 mm (SD 0.17) and 0.61 mm (SD 0.15) and the mean incisal reduction for Groups A, B and C was 1.0 mm (SD 0.28), 1.0 mm (SD 0.38) and 1.03 mm (SD 0.26) respectively. CONCLUSION: It is suggested that consideration be given to the use of a silicone index or depth gauge bur when teeth are prepared for porcelain laminate veneers. PMID- 11048392 TI - The physical mechanisms of complete denture retention. AB - The purpose of this article is to assist the practitioner to understand which factors are relevant to complete denture retention in the light of the current understanding of physics and materials science and thus to guide design. Atmospheric pressure, vacuum, adhesion, cohesion, surface tension, viscosity, base adaption, border seal, seating force and muscular control have all been cited at one time or another as major or contributory factors, but usually as an opinion without proper reference to fundamental principles. Although there has been a detailed analysis published, it seems appropriate that a restatement of the points in a collated form be made. In fact, denture retention is a dynamic issue dependent on the control of the flow of interposed fluid and thus its viscosity and film thickness, while the timescale of displacement loading affects the assessment. Surface tension forces at the periphery contribute to retention, but the most important concerns are good base adaptation and border seal. These must be achieved if full advantage is to be taken of the saliva flow-related effects. PMID- 11048394 TI - Variations in the presenting and treatment features in reimplanted permanent incisors in children and their effect on the prevalence of root resorption. AB - OBJECTIVE: To examine variations in the presentation and treatment of reimplanted incisors in children and to determine the effect of these on the prevalence of external root resorption. SETTING: Departments of Paediatric Dentistry, Belfast and Newcastle upon Tyne. DESIGN: Recording of the timing of the injury and the storage mediums (including air) and of reimplantation, the stage of root development, the degree of contamination and the time of commencement of root treatment. Cases were reviewed clinically and radiographically at intervals of 3 months. Root resorption was classified as present or absent. Logistic regression and cross-tabulations were produced with the presence of resorption set as the outcome. RESULTS: 128 reimplanted permanent incisor teeth, their median dry time prior to reimplantation being 15 minutes (range 4-52 mins), the median time in a liquid medium being 45 minutes (range 0-650 mins), with a median splinting time of 15 days (range 4-52 days) and a median pulp extirpation time of 15 days (range 0-612 days). There was a lower prevalence of resorption when the period of dryness was less than or equal to 5 minutes (p = 0.025). The prevalence of resorption in teeth with no visible contamination was 57.1%, for those with contamination which were washed clean it was 75%, in those rubbed clean it was 87.5%, and it was 100% for those reimplanted with visible contamination still present (p = 0.014). The corrected odds ratio for contamination was 2.99 and for an extension of 10 minutes of dryness it was 1.29. CONCLUSION: The degree of contamination and the period of dryness were the major risk factors for resorption in this study of reimplanted teeth in children. PMID- 11048396 TI - [In Process Citation] PMID- 11048395 TI - Prevalence of long-term use of medicines with prolonged oral clearance in the elderly: a survey in north east England. AB - AIM: To determine the prevalence of long-term use by the elderly of prescribed and over-the-counter (OTC) medicines with prolonged oral clearance (POC), with regard to sugars content, dose form and therapeutic groups of medicines used. DESIGN AND SETTING: Two cross-sectional observational surveys in ten general medical practices in north-east England during 1996. METHODS: Computerised patient records of all elderly patients (aged 60 years and over) were surveyed for prescribed medicines use. Within these practices, 50% of elderly patients registered with ten general medical practitioners were surveyed by postal questionnaire to assess over-the-counter (OTC) medicines use. RESULTS: Of 20,731 elderly patients registered, prevalence of use of prescribed prolonged oral clearance (POC) medicines was 9.8% (95% CI: 8.2%, 11.3%) and use in females aged 75 years and older was significantly more likely (P < 0.0001). Of 2,796 prescribing instances (PIs) for 143 POC medicines used long-term, 53% were gastrointestinal and 72% were sugars-free; however, 82% of 542 PIs for generic liquids were sugars-containing compared with 8% of 685 PIs for proprietary liquid oral medicines. Of 1,532 elderly respondents to a postal questionnaire, 17 were using 13 different OTC medicines with POC regularly and long-term (mean prevalence; 1.1%). Of the 17 instances of regular long-term use of OTC medicines, 59% were sugars-free. CONCLUSIONS: Prescribed medicines represent the bulk of regular, long-term medicines use in the elderly. Generic prescribing is more likely to result in sugars-containing medicines being dispensed. Generic medicines manufacturers must be encouraged to provide sugars-free alternatives to POC medicines used long-term, and health professionals should be vigilant when prescribing and dispensing these medicines to the increasingly dentate elderly. PMID- 11048397 TI - [Recent data on prion diseases]. AB - The transmissible spongiform encephalopathies (TSEs) or "prions diseases", provokes some controversies about their origins and possible transmission from animals to humans. Important advances were obtained recently on the genetical TSEs with the genotypic diagnosis methods based on the discovery of numerous mutations and inclusions in the PRNP gene coding for prion protein. The possible connection between the bovine spongiform encephalopathy (BSE) and the newvariant of Creutzfeldt-Jakob disease (nv-CJD) ask numerous questions in terms of public health. Lastly, the important problem of biological diagnosis of "prions diseases" presents interesting advances that must be continued and supported by the public health authorities. PMID- 11048398 TI - ["Dr. Jacques Forestier (1890-1978), thermal doctor at Aix-les-Bains": some recollections of one of his students]. PMID- 11048399 TI - [In Process Citation] PMID- 11048400 TI - [Medical photography and cinematography before 1914: privileged rapport with the neurosciences]. AB - Arthur Van Gehuchten (1861-1914), Belgian anatomist and neurologist, professor at the Universite catholique de Louvain, was an enthusiastic user of photography and a pioneer of medical cinematography. He used these techniques widely in order to illustrate his lectures, papers and his neurological textbook. His films and photographic plates are at the present time being studied and restored. The nitrate films are the oldest belgian films surviving. Neuroscientists have played a major role in the development of medical photography and cinematography and in the use of these techniques for clinical, research or didactic purposes. PMID- 11048403 TI - [Cardiology at the end of the 20th century]. AB - In autumn 1999 results of two well-controlled studies were published that are consistent with a frequent association between Helicobacter pylori seropositivity and coronary heart disease (CHD). Concerning the therapy of CHD, attention is mainly focused on new thrombolytic agents, bypass grafting (CABG) and balloon angioplasty (PTCA). In patients with intractable angina where aggressive medical therapy was exhausted and who were no longer candidates for CABG or PTCA, transmural laser revascularisation (TMLR), enhanced external counterpulsation (EECP) and spinal cord stimulation can be considered. TMLR was shown to improve symptoms but not myocardial perfusion; the preoperative mortality accounts for 10 20%. In hypertrophic obstructive cardiomyopathy, alcohol-induced transmural septal myocardial ablation (PTSMA) reduces both the symptoms and the left ventricular outflow tract gradient. Although the prevalence of hypertension emergencies has dramatically diminished, the number of hypertensive patients with heart failure and end-stage renal disease is increasing. It is important to detect and treat mild hypertensives in early stages, especially diabetics and younger women with additional risk factors and/or proteinuria. The choice and dosage of drugs is to be individualised. In chronic heart failure (CHF), the protective effect of ACE inhibitors, metoprolol and carvedilol has been repeatedly shown in CHF stage NYHA II and III. The merit of ACE inhibitor and beta-blockers in high doses remains questionable in old patients and those with severe CHF (NYHA IV). In the latter indication, spirolactone was successfully reintroduced. Eplerenone (epoxymexrenone) is a new aldosterone antagonist with little affinity to other steroid receptors and has therefore less undesirable effects than spirolactone. The body of knowledge in therapeutic and technical progress in medicine of the 20th century are summarised and their positive and negative consequences briefly discussed. PMID- 11048402 TI - [Viper (Vipera berus) snake bites]. AB - Cases of snake bites (Vipera berus) have as compared with past years a rising trend in the Czech Republic. This ensues among other factors from a higher prevalence of snakes due to the improving ecological situation. The morbidity of snake bites is of no epidemiological importance, the frequency of snake bites amounts to several tens per year and in some clinically manifest intoxication does not develop. Nevertheless in individual cases, in children weakened subjects, a viper bite may be manifested by a serious and in exceptional instances fatal affection. Within the framework of first aid the authors do not recommend application of a tourniquet or dissecting of the wound because of undesirable potentiation of tissue traumatization. Non-specific treatment involves the administration of corticoids and antihistaminics. Specific immunotherapy, administration of horse antiserum (Ipser Europe, Pasteur Merieux, France) is indicated only in case of systemic or very severe local symptoms and is associated with the risk of a severe allergic reaction. In case of severe systemic symptoms, symptomatic treatment in a health institution of the appropriate type is of fundamental importance. In all cases observation of the affected subject is recommended to rule out intoxication or the development of possible complications. PMID- 11048401 TI - [Th2 lymphocytes in man: a new cause of hypereosinophilic syndrome]. AB - Interleukin-5 produced by Th2-type lymphocytes is involved in the pathogenesis of several hypereosinophilic disorders. We have identified clonal Th2 cells in the peripheral blood of three patients with the idiopathic hypereosinophilic syndrome. Costimulatory signaling through B7/CD28 and LFA-3/CD2 pathways cooperates with an autocrine interleukin-2/interleukin-2 receptor loop for the survival and proliferation of these Th2 cells, as well as their production of cytokines, independently of T cell receptor engagement. The high-level of spontaneous apoptosis displayed by these cells was inhibited by interleukin-2 and interferon-g. New therapeutic strategies could result from our observations. Indeed, the hypereosinophilic syndrome may represent an unexpected indication for new immunomodulatory molecules such as CTLA4-Ig and anti-il-2 receptor antibodies. PMID- 11048404 TI - [Urinary tract infections--still a real problem]. AB - By analyzing the standard works in the scientific world literature, it was possible to construct a contemporary view on the etiology and antibiotic therapy of some urinary tract infections. It can be stated, that the most effective remedies are cotrimoxazol and fluoroquinolones. In contrary to beta-lactam antibiotics, cotrimoxazole and fluoroquinolones work with a rapid bactericidal effect, they have long elimination half-times and better sterilization capacity in the periurethral region, and it use brings minimal risk of recurrence of the infection. This facts are compared with results of bacteriological investigation of urine, which were done in the microbiological laboratories of University Hospital Hradec Kralove, Czech Republic, in 1998. The most frequent origins of urinary tract infections are Escherichia coli and Enterococcus spp. Stains. The are keeping high susceptibility to contrimoxazole, furantoin and oxoline acid. The other Gram negative rods isolated from hospitalised patients are highly resistant to most of antimicrobial agents included fluorochinolons, which resistance culminates to 50%. The widest spectrum of pathogens and the highest resistance was found in the hospitalised patients of the university hospital. High percentages of resistant strains was also in patients from the district hospitals. The results are discussed. PMID- 11048405 TI - [Detection of myeloma cells in peripheral blood using multi-parameter flow cytometry: monitoring residual disease]. AB - BACKGROUND: The aim of this study was to evaluate significance of CD 38(+2)45 54+, CD 38(+2)45-56+ and CD 38(+2)45-138+ cells counts in peripheral blood of patients with multiple myeloma for monitoring of the minimal residual disease. METHODS AND RESULTS: A triple-color flow cytometric analysis was used for this purpose. Peripheral blood of 29 patients with multiple myeloma who underwent high dose chemotherapy and autologous stem cells transplantation was repeatedly analysed. Counts of myeloma cells in peripheral blood were compared to serum monoclonal immunoglobulin concentration, serum calcium level, serum C-reactive protein, serum beta 2 microglobulin, and number of myeloma cells in bone marrow (morphology). From 29 patients in this study, 5 patients have relapsed. Patients in relapse had significantly higher counts of CD 38(+2)45-54+, CD 38(+2)45-56+ and CD 38(+2)45-138+ cells in peripheral blood than patients in remission (geometrical average: 12.41; 6.20; 14.45 cells/microliter versus 4.08; 2.87; 2.58 cells/microliter). The number of these cells correlated well with serum monoclonal immunoglobulin level and counts of myeloma cells in bone marrow. CONCLUSION: The authors conclude that the longitudinal multi-color flow cytometric analysis of CD 38(+2)45-54+, CD 38(+2)45-56+ and CD 38(+2)45-138+ cells in peripheral blood of patients with multiple myeloma is a useful method for evaluation of the disease activity. Significance of peripheral myeloma cells count for prediction of the relapse of the multiple myeloma remains to be evaluated. PMID- 11048406 TI - [Hematologic and cytogenetic response to treatment in patients with chronic myeloid leukemia]. AB - BACKGROUND: The Interferon alpha therapy increases the number of cytogenetic responses in patients with chronic myeloic leukaemia. The addition of cytarabine can reduce the number of Ph positive metaphases. The achievement of cytogenetic response is connected with longer survival of patients with chronic myeloic leukaemia. The aim of the study was the evaluation of the achievement of hematologic and cytogenetic response as well as adverse effects of the treatment in chronic myeloic leukaemia patients. METHODS AND RESULTS: The followed was the group of 87 previously untreated CML Ph positive patients. 34 patients with the median age of 44.6 years were treated with hydroxyurea, 42 patients were treated with single interferon alpha and 11 patients with the median age of 41.3 years with the combined interferon plus cytarabine therapy. The complete hematologic remission occurred in only 17.5% of patients treated with hydroxyurea, but in 35.7% treated with interferon and in 54.5% patients treated with the combined therapy. The cytogenetic response we have not found in any of hydroxyurea treated patients, in the group of interferon alpha in 38%. The highest number of cytogenetic responses was in the group treated with interferon plus cytarabine. As we have expected, the addition of cytarabine increased hematotoxicity and gastrotoxicity. CONCLUSION: Based on the published date, that show a better survival of patients with the achieved cytogenetic response as well as the higher number of cytogenetic responses in the group of interferon plus cytarabine therapy from our observation, we believe, that combined therapy should be suitable as a front-line therapy of chronic myeloic leukaemia patients. PMID- 11048407 TI - [Metamizol--a new effective analgesic with a long history. Overview of its pharmacology and clinical use]. AB - Metamizol is an effective, non-opioid analgesics which was originally introduced to the therapy in the year 1922. However, with the reference to the side effects of other related pyrazolone derivatives its administration, similarly as the usage of other pyrazolones, was significantly limited. Later, metamizol has been used, usually mixed, with spasmolytic agents and quite recently it has been introduced as a mono-component medicament. Metamizol proved to be a very effective analgesic. When administered in equipotent doses, it had its effects comparable to various opioid analgesics, such as tramadol, pentazocine and pethidine. Beside the strong analgesic effect, it produces also significant antipyretic and splasmolytic effects without the adverse, unpleasant anticholinergic impact. Its spasmolytic effect on the smooth muscle of the sphincter Oddi, urinary tract, and the gal bladder is comparable to the effects of buthylscopolamine. Unlike aspirin and other nonsteroidal antiinflammatory drugs it has, however, no antiinflammatory activity when administered in clinical doses. Similarly, metamizol has no effect on the CNS, cardiovascular system, renal and metabolic functions. On the other hand, metamizol, like aspirin, has got a significant effect on the aggregation of platelets. Metamizol is basically a prodrug. The parent substance is not effective before its conversion into two active metabolites (4-methylaninoantipyrine and 4-aminoantipyrine) in the body. Metamizol is well absorbed from the small intestine but only two above mentioned active metabolites and no parent drug can be detected in the blood. The active metabolites are consequently metabolised to ineffective metabolites including the relevant acetylderivatives, in which the acetylation phenotypes can be distinguished. In the therapy, metamizol can be used, as an analgesic, at post surgical pain, patient's controlled analgesia (PCA), at the cancer's pain and in the pains of different origin (post-traumatic pain, the pain at myocardial infarction, craniocerebral trauma, and the invasive diagnostic interventions), as well as at he pain of neuromuscular origin, headache and migraine. Its spasmolytic effect in connection with a strong analgesic activity is very useful at various colic attacks. Further, metamizol is a useful antipyretic both in the adults and children. Its adverse effects are not pronounced and drug interactions are minimal. Only when metamizol is administered together with cyclosporine, the blood levels of the last substance should be regularly checked. PMID- 11048408 TI - [Multihormonal and multifunctional hypophyseal adenoma and the acromegaly syndrome]. AB - Woman 75-year-old treated 30 years for syndrome of acromegaly refused pituitary surgery and irradiation. Five years and nine months before death she had a colon carcinoma successfully removed. Multinodular hyperfunctional goitre was treated with carbimazole. For six last years of life corticosteroids were given as a replacement therapy. Her cause of death was the heart failure due to acromegalic heart disease. In autopsy a large intrasellar and extrasellar pituitary adenoma without rests of nonneoplastic tissue was found. Nevertheless the target peripheral endocrine glands except ovaries, were not atrophic. A multinodular goitre and diffuse adrenocortical hyperplasia were revealed. Histology, and immunohistochemistry demonstrated that mot neoplastic cells were producing GH and ACTH, dispersly Prl, scattered cells were positive for beta-subunit of FSH, LH, TSH. Electron microscopy proved most of the cells to be densely granulated. We classify the adenoma according to the newly proposed WHO pituitary tumours classification (1) as plurihormonal, hyperfunctional, extrasellar, typical adenoma from densely granulated cells. We conclude that in plurihormonal adenomas with dominant (in the case referred acromegalic) symptomatology the additional hormonal production should be monitored as a possible source of important complications. PMID- 11048409 TI - [Monochorionic twins after treatment of sterility using assisted reproduction methods]. AB - OBJECTIVE: The goal of this study has been to determine frequency of occurrence of monochorial twins within the study group consisting of 521 pregnancies conceived through the In Vitro Fertilization (IVF) programme. DESIGN: An observational study. SETTING: Sanatorium Pronatal, Na Dlouhe mezi 4/12, 147 00 Praha 4-Hodkovicky. METHODS: A condition to be included in the study group was that there was a gestation sac detected by ultrasound. The first ultrasound examination was performed transvaginally and was done between the fifth and the sixth week of each pregnancy. A multiple pregnancy has been classified as monochorial in case when gestation sac contains two yolk sacs and two fetuses. RESULTS: Within the whole study group there were 13 monochorial twins which presents 2.5% of all pregnancies. The incidence of monochorionicity among all multiple pregnancies is 6.4%, in IVF cycles without micromanipulation techniques the occurrence of monochorial twins has been 0.9%, following cryoembryotransfer 3.9%, with micromanipulation (ICSI, AH) it reaches 5.7% of all pregnancies. CONCLUSION: Monochorial twins were detected seven time more frequent in comparison within occurrence in ordinary population. The occurrence rises in connection with the number of fetuses in uterus. In comparison with simple IVF cycles we have proved higher occurrence of monochorionicity in connection with micromanipulation techniques and cryoembryotransfer. PMID- 11048410 TI - [Heterotopic pregnancy and its occurrence in assisted reproduction]. AB - OBJECTIVE: To assess the incidence of heterotopic pregnancy after infertility treatment using the technology of assisted reproduction. DESIGN: A prospective study of 618 women who became clinically pregnant following assisted reproduction technology (ART) procedures. SETTING: Sanatorium Pronatal, Na dlouhe mezi 4/12, 147 00 Praha 4-Hodkovicky. METHODS: Study group consists of clinical pregnancies conceived after ART procedures within the period from January 1, 1997 until June 30, 1998. A condition to be included in survey group was that there was a gestation sac detected by ultrasound or histological confirmation of ectopic pregnancy. RESULTS: Six-hundred-eighteen clinical pregnancies resulted and 23 of the pregnancies were ectopic gestations (3.7%). Seven out of the 23 (30.5%) ectopic pregnancies were heterotopic. Thus heterotopic pregnancy rate after ART was, 1.14% (1 in 88). CONCLUSION: The incidence of heterotopic pregnancy following assisted reproduction technique is relatively frequent. This condition represents a live-threatening complications of pregnancy. The prognosis for intrauterine gestation in case of heterotopic pregnancy is usually good. About 78% delivered living child at term. PMID- 11048412 TI - [Effect of the number of transferred embryos on multiple pregnancy]. AB - OBJECTIVE: The aim of this retrospective study was to assess the influence of the number of transferred embryos on multiple pregnancy. DESIGN: Retrospective clinical study. SETTING: Department of Obstetric and Gynecology FN UP, Olomouc. METHODS: Over the last seven years 572 embryotransfers (ET) have been performed in our centre. RESULTS: Of the 173 pregnancies (P) achieved there were 56% singletons, 35% twins and 9% triplets. One embryo was transferred in 10% of cases and all 8 pregnancies (15% P/ET) ended with a live birth. Two embryos were transferred in 17% of cases. The pregnancy rate was 20% P/ET, 55% of pregnancies were twins and 10% pregnancies were lost in abortion or ectopic pregnancy. Three embryos were transferred in most cases (62%). The pregnancy rate was the highest in this group (36% P/ET) with 13% twins and 3% triplets. The overall pregnancy loss was 16%, the highest loss, 27%, recorded in triplets. Four embryos were transferred in 11% of cases. The pregnancy rate decreased to 28% P/ET, as well as the number of twins (6%), while the number of triplets increased slightly (5%). Pregnancy loss occurred only in singleton pregnancies. CONCLUSION: We can conclude that in our setting the pregnancy rate increases with a higher number of embryos up to three embryos transferred. Unfortunately, the higher pregnancy rate with three embryos transferred is on the expense of a higher risk and losses due to multiple pregnancy. Therefore we recommend to transfer two embryos unless there are some specific reasons. PMID- 11048413 TI - [Screening for Down's syndrome in the 10th to 13th week of pregnancy]. AB - OBJECTIVE: The implementation of antenatal screening for Down's syndrome at 10 13. week using nuchal translucency, maternal serum free subunit of human chorionic gonadotrophin and the mother's age. DESIGN: Nuchal translucency scan (NT) and serum free beta human chorionic gonadotrophin subunit (FB hCG) were measured at 283 consecutive pregnant women booked at the same antenatal care facility between 10-13. gestational week. SETTING: Antenatal screening center Klimentska, Prague. METHODS: Medians of both markers according to gestational age as well as retrospective combined risk of Down's syndrome were assessed. RESULTS: 95 per cent of NT measurements was below 1.77 multiples of median. 95 per cent of FB hCG measurements was below 1.88 multiples of median. Combined risk higher than 1/300 had 1 per cent mothers (total 3 screen positive results). CONCLUSION: The results of this pilot study serve for implementation of screening for Down's syndrome at 10-13. week as well as for integration of both first and second trimester screening systems. PMID- 11048411 TI - [Biochemical screening in the second trimester of pregnancy from a genetic aspect]. AB - OBJECTIVE: Evaluation of the efficiency of detection of inborn chromosomal aberrations by biochemical screening of pregnant women. DESIGN: Summary of 9 years study by karyotyping of amniocytes of pregnant women, when pathological levels of AFP, hCG, uE3 were detected. Ethical problems of genetic counselling of pregnant women. SETTING: Department of Medical Genetics FTN, Chair of Medical Genetics IPVZ, Prague 4. METHODS: Biochemical screening of pregnant women in II. trimester of pregnancy by Triple Test and computer programme Prenatal Software 1.2.K, cytogenetical examination of amniocytes from amniotic fluid after amniocentesis in II. trimester. RESULTS: Among 6,471 pregnant women tested by Triple Test 20% had abnormal levels of test. In 1.5% of them an abnormal fetal karyotype was detected. Only 1/3 of them were Down Syndromes, we found also other types of aberrations. During ethically suitable counselling only 2/3 of pregnant women decided to terminate the pregnancy. CONCLUSION: Biochemical screening of pregnant women in II. trimester is very important help for detection of inborn chromosomal aberrations. The women must be informed by suitable ethical approach. PMID- 11048414 TI - [Antibiotic prophylaxis of infectious complications in cesarean section- prospective study]. AB - OBJECTIVE: To verify the significance of Cefazolin administration to women who were indicated for caesarean section. SETTING: Department of Obstetrics and Gynecology, First Faculty of Medicine, Prague, Czech republic. METHODS: Cefazolin was administered to 30 women with a patient history risk in a dose of 1 mg i.v. after ligation of the umbilical cord. The control group consisted of 30 women. Observed parameters: weight gain during pregnancy, indications for caesarean section, culture from the cervix and vagina prior to the caesarean section, culture from the amniotic fluid, culture from the 4th day lochia, temperature curve, leucocyte count before and 4 days after the operation, complications after the surgery, administration of antibiotics, and length of hospitalisation. RESULTS: In the group with antibiotic prophylaxis, we observed a more favourable course of the temperature curve and a smaller leucocyte count the fourth day after the surgery (p < 0.01), in comparison with the women without prophylaxis. The duration of hospitalisation in women with antibiotic prophylaxis was one day shorter and there was no occurrence of febrile complications requiring further antibiotic treatment. CONCLUSION: Cafazolin is advantageous antibiotic for prophylaxis in caesarean section in patients with risk. PMID- 11048415 TI - [Diagnosis and treatment of chronic vaginal candidiasis]. AB - OBJECTIVE OF STUDY: The objective of the study is to determine the optimum diagnostics of vaginal yeast infections and to compare the effects of treatment of these infections by Natamycin with those by Clotrimazol. TYPE OF STUDY: Prospective comparison of both option of treatment of vaginal infections. NAME AND PLACE OF RESEARCH: Obstetrics and gynaecology department, Brno-Bohunice. METHODS: 30 patients treated with hydrophobe Natamycin and 20 patients treated with hydrophyll Clotrimazol formed a sample of 50 women. Regular checks were made on the 10th and 30th day after the beginning of treatment. Diagnosis was performed by means of native microscopy supplemented by an examination for cultivated yeasts in the culture medium "FUNGI-QUICK". At the same time a microscopic examination of slides stained by Gram and Giems was made. RESULTS: A correlation between the evaluated native slide and the culture examined thereafter was 96%. Statistical evaluation of the difference of the rate of success of treatment between the two groups by means of the t-test revealed a value of 0.29, the level of probability was 0.05 at N1 = 0.1795 and N2 = 0.2179. CONCLUSION: Native microscopy is irreplaceable in the diagnosis of vaginal candidosis. No significant differences in the effects of treatment with Natamycin and Clotrimazol were found. On the basis of these results we made some recommendations on the principles of optimum treatment. PMID- 11048417 TI - [The definition of laparoscopic hysterectomy]. AB - OBJECTIVE: To define laparoscopic hysterectomy. DESIGN: Short review. SETTING: Departments Gynaecology, Kladno Hospital and Na Homolce Hospital, Prague. METHOD: A retrospective study and analyse of literature and information database (Medline 1994-1998). CONCLUSION: We suggest nomenclature and LH classification for the Czech Endoscopic Society, which will bases on the guidelines published by Nezhat et al. and defined especially two forms of laparoscopic assisted vaginal hysterectomy (superior and inferior type). PMID- 11048416 TI - [Preperitoneal laparoscopic retropubic colposuspension in the treatment of stress incontinence: the mesh and tacker technique]. AB - The Burch procedure has enjoyed in the last decade a favourable status among open surgical repairs for stress urinary incontinence. In the last few years this technique was adapted for endoscopic application. This results in decreased recovery time and diminished postoperative patients discomfort. This endoscopic procedure was further simplified by means of extraperitoneal approach and through the use of Mesh & Tacker technique. This enables a high quality durable colposuspension in significantly shortened operational time. OBJECTIVE: Evaluation of preliminary results and experiences of the above-mentioned new laparoscopic extraperitoneal approach in the treatment of stress incontinence. SETTING: Department of Gynaecology and minimally invasive surgery Na Homolce Hospital. DESIGN: Prospective pilot study. METHODS: The patients with stress incontinence proven clinically and by means of urodynamic investigation (cystometry, stress profilometry and uroflowmetry) were included in the study. The Retzius space was dissected laparoscopically via preperitoneal distention balloon (PDB, Origin Medsystems). Colpofixation to Coopers ligaments was achieved by means of Mesh & Tacker technique, e.g. polypropylen Mesh and aplicator of helicose spirals (Origin Medsystems). RESULTS: In this pilot study of initial 8 patients we may conclude that the above-mentioned method is promising. Of note is shortened operational time (35 min), easy performance without necessity of laparoscopic endosuturing and minimal tissue damage. The small amount of patients and short follow up period would not allow definite conclusions but all the patients are sofar fully continent. PMID- 11048418 TI - [Bacterial zoonoses: listeria infection of the mother in the 3rd trimester of pregnancy]. AB - Listeria species are Gram-positive bacilli, present in food and in the environment (soil, water). Most of them are fortunately low-virulent, so there is no risk for healthy people. However, in special circumstances, they can be pathogenic, in immunocompromised individuals. Septicaemia in pregnancy is not dangerous for women, but for the foetus, so delayed diagnosis will lead to spontaneous abortion, intrauterine death or eventually to premature labour. We report in our paper case of Listeria septicaemia in a pregnant woman in the third trimester of the pregnancy. Thanks to early diagnosis and subsequent causal antibiotic treatment, we were able to prevent the development of congenital infection and a healthy baby was born. PMID- 11048419 TI - [Guidelines for malignant gynecologic tumors. I. Standard-primary comprehensive treatment of malignant epithelial tumors. Oncologic gynecology section of the Czech Gynecologic and Obstetrical Society]. PMID- 11048420 TI - [Assisted reproduction in the treatment of immune-related infertility]. PMID- 11048421 TI - [Recommendations for methods of preinduction and induction of labor]. PMID- 11048422 TI - [Contributions of the Society of Clinical Cytology to the history of the Czech Medical Society]. PMID- 11048423 TI - [Response to article by Klimova K., Maly Z.: Indications and risks of invasive methods in prenatal diagnosis--who indicates and who bears the risk?]. PMID- 11048424 TI - [Use of the Citation Index and impact factors]. AB - Article deals with evaluation method of scientific works by means of Citation Indexes and Impact Factor which are produced by Institute for Scientific Information of Philadelphia. It criticises criteria of the usage of Impact factor and way of research workers evaluation. PMID- 11048425 TI - Outbreaks of Salmonella typhimurium DT204b infection in England and Wales and elsewhere in Europe. PMID- 11048426 TI - Active surveillance of rare and serious diseases in children. PMID- 11048427 TI - AIDS and HIV infection in the United Kingdom: monthly report. PMID- 11048428 TI - Some in vitro invasion inhibition of red cells by in vivo nonprotective anti-LDH antibodies of Plasmodium berghei. AB - In Plasmodium berghei, sephadex G-200 purified lactate dehydrogenase (LDH) fraction immunized mice did not exhibit protection when challenged with 1 x 10(6) P. berghei-parasitized erythrocytes. However, LDH immunized mice seroconverted and showed an antibody titre of 1:2048 by indirect haemagglutination (IHA) and 1:160 by indirect fluorescent antibody (IFA) assays. Fluorescence was distributed evenly on P. berghei-parasitized red cells showing no specific location of parasite LDH. Anti-LDH antibodies supplemented in 19 h in vitro culture of P. berghei exhibited 9.2% invasion inhibition into the fresh red cells. PMID- 11048429 TI - The pathology of goat paratuberculosis: gross and histopathological lesions in the intestines and mesenteric lymph nodes. AB - From a pathological examination of the intestinal tracts of 1590 goats killed at slaughterhouses in the Fars Province of Iran, 59 cases (3.71%) were suspected, on gross examination, of having paratuberculosis. The diagnosis was confirmed by histopathological study and Ziehl-Neelsen staining of direct smears of rectal faeces. On the basis of severity of involvement of the terminal ileum and mesenteric lymph nodes, the microscopic lesions were classified to mild, moderate and severe forms. Caseous necrosis and calcification were observed only in the mesenteric lymph nodes. High numbers of acid-fast organisms were present in the epithelioid macrophages of the intestine but were inapparent or sparse in the mesenteric lymph nodes. On microscopic examination, 13.5% of the suspected animals were found to have paratuberculosis, in comparison with 3.38% by direct faecal smears. In addition, 30.5% and 15.3% of the animals were diagnosed as having eosinophilic enteritis and linguatulosis, respectively. These findings stress the importance of a careful histopathological examination of the intestines and mesenteric lymph nodes for the diagnosis of paratuberculosis in goats. PMID- 11048430 TI - Kinetics of antibody production and clinical profiles of calves experimentally infected with Listeria monocytogenes. AB - Clinical and serum antibody profiles were studied during oral Listeria monocytogenes infection of calves. No clinical signs, except for pyrexia with mild diarrhoea and staggering gait, were observed in the infected calves. Specific antibodies to listeriolysin O (LLO) appeared as early as day 8 of an oral infection and peaked by days 16-32 of infection. Antibodies to LLO were observed to persist over the period of 126 days observed in the study. LLO being a major virulence factor and capable of inducing a humoral response could therefore be used as an antigen for development of an immunoassay for diagnosis of Listeria monocytogenes infections in animals. PMID- 11048431 TI - Serological investigations of red foxes (Vulpes vulpes L.) for determination of the spread of tick-borne encephalitis in Northrhine-Westphalia. AB - Serum samples from 786 red foxes shot between January 1995 and August 1996 in the southern half of Northrhine-Westphalia, located in western Germany, were tested for the presence of antibodies against tick-borne encephalitis (TBE) virus using the Immunozym FSME IgG All Species-ELISA (Immuno, Heidelberg, Germany) as a screening test: 759 sera were negative, 23 (2.9%) were borderline, and four (0.5%) were positive. Nine of the 27 ELISA reactive sera were confirmed by the TBE Western-Blot (Immuno, Heidelberg, Germany). Furthermore these 27 sera were tested for neutralizing antibodies by means of a plaque reduction neutralization test (PRNT) against TBE and West Nile viruses. Only one single serum was found to have a neutralization titre (+1:800 PRNT80) against TBE virus. All other 26 sera were negative for neutralizing antibodies against TBE or West Nile virus. Since the titre of the single serum is low, it can be interpreted that if TBE virus is present, its prevalence is extremely low. Northrhine-Westphalia is not classified as a TBE-endemic area. Further calculated serological testing of game and virological investigation of collected ticks in the affected area seem to be meaningful and necessary. PMID- 11048432 TI - Polluting profiles of dieldrin and DDTs in laying hens of Osaka, Japan. AB - Contaminating/accumulating levels of organochlorine pesticides in extractable fats from a basal diet, eggs and seven tissues (adipose tissue, blood, kidney, liver, muscle, ovary and oviduct) and excreta of laying hens that were kept in a general poultry farm of Japan were examined. No benzene hexachlorides or aldrin was detected [< 1 part per billion (p.p.b.)] overall. Dieldrin and all DDTs (p,p' DDE, o,p'-DDT, p,p'-DDD and p,p'-DDT) were contaminated in the dietary fats (mean range 10-70 p.p.b.). Although dieldrin (4-58 p.p.b.), p,p'-DDE (65-196 p.p.b.) and p,p'-DDT (30-73 p.p.b.) were found to be accumulated in all the tissue fats and egg yolk fats, they were not detected in the dried excreta. Accumulations of o,p'-DDT and p,p'-DDD were found only in the liver fat (92 p.p.b.) and in the kidney fat (27 p.p.b.), respectively. In all the samples, p,p'-DDE levels were highest in comparison with the other compounds. For all organochlorine pesticides detected, the accumulated levels were well below the practical residue limits. PMID- 11048433 TI - The ability of the enzyme-linked immunosorbent assay to detect antibody against Staphylococcus aureus in milk following experimental intramammary infection. AB - Changes in the milk antibody levels against Staphylococcus aureus were measured at the start of an experimental intramammary instillation of either S. aureus (Study I) or Staphylococcus hyicus (Study II). A commercial enzyme-linked immunosorbent assay system was used. Twenty-one Holstein cows were enrolled in Study I and 15 Holstein cows were used in Study II. Pathogen instillation began 21 days before the start of the non-lactating period. Cows received intramammary antibiotic treatment in all quarters immediately after the last milking, the start of the non-lactating period. Lacteal secretions were collected before the start of the non-lactating period, and during the immediate postpartum period in both studies, and during the non-lactating period in Study I. Milk was cultured for mastitis pathogens and S. aureus antibody levels and somatic cell counts were determined from all samples. There was an approximate 2-week delay in the elevation in antibody levels in response to the instillation of S. aureus. Antibody levels remained elevated in cows with S. aureus intramammary infections postpartum, but were below threshold in cows where intramammary infections were cured during the non-lactating period. Antibody levels were elevated by S. hyicus intramammary infections, remained elevated for the first 12 days postpartum, but were below threshold by day 21 postpartum. Cows with incipient intramammary S. aureus infections might be misclassified as false negatives by the antibody test. However, results suggest that cows with S. hyicus intramammary infections that were not cured would not be misclassified if milk is withheld from test for the first 30 days postpartum, as recommended by the manufacturer of the test. PMID- 11048434 TI - Protective effect of vaccination with culture supernate of M. hyopneumoniae against experimental infection in pigs. AB - The protective activity of Mycoplasma hyopneumoniae inactivated vaccine prepared from sedimented whole cells and cell-free culture supernates was evaluated experimentally using hysterectomy-produced, colostrum-deprived pigs in which mycoplasmal pneumonia had been induced. The culture supernate vaccine containing less than 10(1) colour-changing units (CCU)/0.2 ml of M. hyopneumoniae significantly (P < 0.05) reduced the percentage of lung lesions compared to controls (3.2 +/- 3.9 vs. 12.2 +/- 2.2%), whereas the sedimented whole cells vaccine containing 10(10) CCU/0.2 ml of organisms provided variable protection (18.7 +/- 16.5 vs. 12.2 +/- 2.2%). Serum from the pigs vaccinated with culture supernate reacted with six protein bands of 97, 89, 65, 46, 42 and 41 kDa by immunoblot analysis. From these results, we conclude that vaccination with culture supernate of M. hyopneumoniae can provide protection against M. hyopneumoniae infection and that these antigens in the culture supernate may be closely related to the reduction of lung lesions. PMID- 11048435 TI - High mortality rate associated with bovine respiratory syncytial virus (BRSV) infection in Belgian white blue calves previously vaccinated with an inactivated BRSV vaccine. AB - In a group of 60 Belgian White Blue calves less than 8 months old still housed in barns, a bovine respiratory syncytial virus (BRSV) outbreak was revealed on the basis of a direct diagnosis (immunofluorescence and virus isolation) performed on the lungs of dead animals, and the kinetics of BRSV neutralizing antibodies. Clinical signs, macroscopical and microscopical pulmonary lesions were also compatible with a BRSV infection. This outbreak is peculiar because the 35 oldest calves (204 +/- 29 days old) had been vaccinated 3-4 months before with an inactivated BRSV vaccine and 30% of these animals had died of respiratory distress. While they experienced a mild respiratory symptomatology, no death was recorded among the 25 youngest calves (69 +/- 29 days old) which had been left unvaccinated. Another peculiarity was found at the histological level where a massive infiltration of eosinophils was demonstrated in the pulmonary tissues of the dead animals. Together these data parallel the dramatic story described 30 years ago in children previously vaccinated with a formalin-inactivated human RSV (HRSV) vaccine upon a natural HRSV challenge. This illustrates that an immunopathological phenomenon also takes place after BRSV vaccination in cattle. PMID- 11048436 TI - Flow cytometry for the detection of enterohaemorrhagic Escherichia coli O157:H7 with latex beads sensitized with specific antibody. AB - To detect low concentrations of enterohaemorrhagic Escherichia coli O157:H7 rapidly, flow cytometry (FCM) was carried out with specific IgG-sensitized latex beads (IgG-Lx). It was found that test samples for FCM can be prepared for much shorter periods by culturing E. coli O157:H7 in trypto-soya broth at 42 degrees C and by treatment with 0.5% formalin at 37 degrees C. FCM with IgG-Lx performed with E. coli O157:H7 prepared by such a procedure revealed that the lowest number of E. coli O157:H7 prepared in pure culture detected by FCM was 10(3)/ml. Because similar findings have already been reported by FCM with immunomagnetic beads, FCM with IgG-Lx is also suggested to be a valuable technique to detect low numbers of E. coli O157:H7 rapidly in food stuffs. PMID- 11048437 TI - [Postoperative results under the new stage classification of lung cancer: the additional reports for those of JACS in 1996]. AB - This time, in 3008 lung cancer patients, the postoperative results were analyzed under the new stage grouping of TNM classification. All of those patients underwent the operation in 1989, and the 5 year-survival rates had beeb surveyed in 1996 by JACS (The Japanese Association for Chest Surgery). Under the new TNM classification established in 1996 worldwidey, T3N0M0 was transferred from IIIA to IIB. This report is the additional one in the focus of the results accompanied with the change of TNM classification. PMID- 11048438 TI - [Prognostic assessment of resected lung cancer based on the new international staging system: analysis by histologic types]. AB - The purpose of this study is to assess the validity of the new international staging system for lung cancer revised in 1997 and to clarify the prognostic value of histologic type. Of 1,042 patients with primary lung cancer who underwent resection from 1982 to 1995, 549 patients with adenocarcinoma (AD) and 363 with squamous cell carcinoma (SQ) were included in this study. Overall deaths including operative deaths were treated as the terminal event. For patients with AD, 5-year survival rates were 83% in T1N0M0, 70% in T2N0M0, 46% in T2N1M0, 45% in T3N0M0, 43% in T1N2M0, 32% in T1N1M0, 27% in T2N2M0, 25% in T3N1M0, and 10% in T3N2M0, respectively. For patients with SQ, 5-year survival rates were 80% in T1N0M0, 78% in T1N1M0, 67% in T1N2M0, 60% in T3N0M0, 56% in T2N0M0, 47% in T2N1M0, 36% in T3N1M0, 26% in T2N2M0, and 10% in T3N2M0, respectively. Survival of T1N2M0 was significantly better than that of T2N2M0 and similar to that of T3N0M0 and T2N1M0 both in AD and SQ. In AD survival was dominantly affected by N factor, while in SQ survival was strongly correlated with T factor. In conclusion, we propose that T1N1M0, T2N1M0, T3N0M0 and T1N2M0 should be classified as stage II and T3N2M0 be included into stage IIIB for AD. The TNM grouping for SQ is same as that for AD except setting T1N1M0 as stage IB and T2N0M0 as stage II. PMID- 11048439 TI - [Evaluation of new TNM classification for lung cancer, especially T3N0M0, stage IIIA, stage IIIB, and pm]. AB - The purpose of this study was to evaluate the results of new TNM staging system for lung cancer in 1997, especially T3N0M0, stage IIIA, stage IIIB, and pm. Five year survival rates of the patients with stage IIIA and stage IIIB were 16% and 18% respectively (NS). Five-year survival rates of patients with T3N1M0, T1N2M0, T2N2M0, and T3N2M0 were 40%, 28%, 15%, and 3%, respectively. The prognosis of T3N2M0 was significantly worse than that of T3N1M0, T1N2M0, and T2N2M0. Five-year survival rates of the patients excluding pm 1 with T4N0M0, T4N1M0, T4N2M0, and T4N3M0 were 21%, 10%, 10%, and 0%, respectively. The prognosis of the patients with T4N0 was significantly better than that of T4N2 and T4N3. In the patients with pm, 5-year survival rates of the patients with pm 1 and pm 2 were 26% and 7%, respectively (p < 0.01). In the patients with pm 1, 5-year survival rates of the patients with N0 + N1 and N1 + N2 were 53% and 16%, respectively (p < 0.01). From our these results, we supported the new TNM system as putting T3N0M0 to stage IIB, putting pm 2 into stage IV. We proposed; 1) chest wall invasion with bone destruction stay in stage IIIA or is T4, 2) T3N1M0 is classified with stage IIB, 3) main stem bronchus invasion is classified with T2, 4) pm 1 is subdivide by N status. Furthermore, stage III seemed to be reasonably subdivided into T1 2N3M0, T4N0-1M0 as stage IIIA and T3-4N2, T1-4N3 as stage IIIB. PMID- 11048440 TI - [Validity and controversies in the new postoperative pathologic TNM classification based on the results of surgical treatment of non-small cell lung cancer]. AB - In 1997, the latest revision of the International System for Staging Lung Cancer was published. To validate the new pathologic TNM classification for non-small cell lung cancer (NSCLC), we analyzed the survival data of 455 patients who underwent pulmonary resection and pathologic staging at our institution from January 1980 through December 1999. The overall 5-year survival rate was 51.0%. Using the revised new stage classification, the survival rate for each stage was as follows; IA: 74.2%, IB: 66.4%, IIA: 56.0%, IIB: 51.8%, IIIA: 21.0%, IIIB: 16.0%, and IV: 0%. The current TNM classification well reflected the long-term prognostic hierarchy. There were significant differences in survival rates between patients with stage IA and IB, and between patients with stage IIB and IIIA. However, there was no significant difference between patients with stage IIA and IIB. No significant difference in survival was observed among patients with stage IIIA, stage IIIB, and stage IV. Five-year survival rate of 48.3% in the T3N0M0 category was significantly better than that of 21.0% found in the new stage IIIA. The survival of patients with intrapulmonary metastases in the same lobe (pm1) was not significantly better than that found in the stage IV. The TNM staging system accurately reflects the prognosis in NSCLC, but some stage definitions can be discussed. PMID- 11048441 TI - [Assessment of the new TNM classification for resected lung cancer]. AB - To evaluate the revised TNM classification, we investigated the prognoses of 552 consecutive patients who had resection of non-small-cell lung cancer between April 1982 and March 1996. According to the new classification, the 5-year survival rate was 76.9% for stage I A, 57.2% for stage I B (I A versus I B, p < 0.0005), 47.7% for stage IIA, 49.8% for stage IIB, 18.6% for stage IIIA (IIB versus IIIA, p = 0.005), 16.7% for stage IIIB, and 7.9% for stage IV (IIIB versus IV, p = 0.02). Especially for patients in stage I A, there was significant difference in survival between patients with the tumor size within 1.5 cm and those with larger than 1.5 cm. The survival rate for T3N0M0 patients was significantly better than that for T3N1-2M0, but there was no significant difference between patients with T3N0M0 disease and those with T2N1M0 disease. Concerning the pm1 patients, the survival rate was significantly better than other stage IIIB patients. Our results supported the revision for dividing stage I and putting T3N0M0 into stage IIB. However, the classification is controversial about dividing stage II and putting pm1 as T4 disease. Furthermore, subgrouping of T1N0M0 disease by tumor size, T3 by tumor invaded organ will be necessary in the next revisions. PMID- 11048442 TI - [Prognostic assessment of the new UICC TNM classification for resected lung cancer]. AB - To evaluated the new UICC TNM classification, we investigated the prognosis of patients who had resection of non-small cell lung cancer. A total of 670 patients with non-small cell lung cancer underwent complete resection and pathologic staging of the disease from 1987 to 1994. The survivals were calculated with Kaplan-Meier methods on the basis of overall deaths, and the survival curves were compared by Logrank test. The 5-year survival rates were 84.6% in stage I A (n = 187), 65.2% in stage I B (n = 177), 41.5% in stage IIA (n = 24), 46.7% in stage IIB (n = 100), 25.6% in stage IIIA (n = 139), 25.8% in stage IIIB and 0 in stage IV. There were significant differences in survival between stage I A and stage I B as well as between stage IIB and stage IIIA. However, there were no significant differences in survival between stage IIA and stage IIB, between stage IIIA and stage IIIB. No significant difference in survival was observed among patients with T1N1M0, T2N1M0 and T3N0M0 (43.9%). In stage IIIB, the patients with pm1 N2 disease (8.9%) had more poorly prognosis than the patients with pm1N0 disease (70.1%) and pm1N1 (38.9%) disease. We concluded that the dividing stage I into A and B categories and placing T3N0M0 in stage II and placing pm2 in stage IV were adequate. In the patients with satellite tumors within the primary lobe of the lung, we think that a new category depended on the N-category is necessary. PMID- 11048443 TI - [Evaluation of TNM classification for lung carcinoma with satellite nodules in the same lobe as the primary]. AB - We conducted a validation of the treatment of satellite nodules in the UICC TNM classification of 1997. Over the past 17 years (1981 to 1997), 29 patients underwent complete pulmonary resection for primary lung cancer accompanied by satellite nodules in the same lobe as the primary. All these patients were categorized as stage IIIB according to the current staging system. The five-year survival rate of all patients was 40% and the figure was unduly better for stage IIIB patients. The three-year survival rate of 20 patients having lymphatic involvement were 18%, but those of 9 patients without it was 89%. Outcome was significantly influenced on their presence or absence of lymph node metastases. The current staging system for patients with satellite lesions in the same lobe appears to be unacceptable because they provide the different influences on prognosis and selecting treatment modality according to the N status. PMID- 11048444 TI - [The validity and problems of a new staging system for lung cancer from the point of view of surgical results]. AB - The validity and problems of the new staging system for resected lung cancer were assessed. In the new staging system for primary lung cancer, stage I is divided into two groups (A and B) on the basis of the tumor diameter. The boundary is 30 mm in maximum diameter of the tumor. On the other hand, intrapulmonary satellite nodules in the resected lung (PM1) are considered as T4. This means that a patient with PM1 is classified as stage IIIB or IV. The five-year cumulative survival rates of 191 consecutive patients of T1 or T2N0M0 who had undergone complete lung resection were calculated. The patients were divided two groups on the basis of the boundaries of the tumor diameter, such as 15, 20, 25, 30 and 40 mm (The tumor diameter in group A is the boundary and less than boundary and the diameter in group B is more than the boundary), and the cumulative survival rates of the two groups were compared. The 5-year cumulative survival rate (Kaplan Meier survival rate) of each group and the p-value (logrank test) were 85.0%, and 0.463 in the case of the boundary of 15 mm; 84.8%, 79.9%, and 0.374 in the case of 20 mm, 82.3%, 80.0%, and 0.553 in the case of 25 mm, 79.5, 83.5, 0.524 in the case of 30 mm and 81.0%, 82.2%, and 0.783 in the case of 40 mm. In all cases, there were no significant differences between the rates in the two groups. On the other hand, 15 cases of T4N0M0 lung cancer included 12 cases of pm1, 2 cases of p3 and 1 case of d2. The five-year cumulative survival rate for T4N0M0 patients with pm1 was 71.1%, which was similar to the rate for stage I or II patients, while the two T4N0M0 patients with p3 died of recurrences of lung cancer at 10 and 13 months after the operation, and the T4N0M0 patient with d2 died of local recurrence of lung cancer 60 months postoperatively. We concluded that the sub classification of stage I on the basis of the boundary of tumor diameter was meaningless, if the boundary was between 15 and 40 mm and that N0M0 patients with PM1 should be classified as stage II or less, or as 1 grade up of T status. PMID- 11048445 TI - [Three approaches to surgical treatment of traumatic disruption of the thoracic aorta]. AB - Traumatic disruption of the thoracic aorta is said to occur most often near the aortic isthmus because of the mechanisms of aortic injury. Between November 1990 and August 1999, we encountered eight cases of surgical treatment for traumatic injury of the thoracic aorta combined with multi-system injury. In some cases, the injury was located near the aortic isthmus; in such cases, we selected surgical options that made use of three different approaches, namely, media sternotomy, posterolateral left thoracotomy, and anteroaxillal thoracotomy. Each approach has advantages and disadvantages. In selecting an appropriate approach, it is not only necessary to consider the various features of the approach itself, but it is also necessary to consider other factors, such as the assisting apparatus in use, the effects of other injuries sustained by perioperative positioning, safety measures against accidental bleeding during surgery, deployment of the operative field, and potential complications after surgery. PMID- 11048446 TI - [Modified Konno procedure for a child with left ventricular outflow tract obstruction after repair of atrioventricular septal defect: a transaortic transpulmonary approach]. AB - A one-year-old infant underwent repair of atrioventricular septal defect with common orifice. About 2 years later, echocardiography revealed a left ventricular outflow tract obstruction for the first time. Because of progression of the obstructive lesion, a modified Konno procedure through a transaortic transpulmonary approach was later performed at 8 years old, and the postoperative course was uneventful. This is a useful procedure for left ventricular outflow tract obstruction with normal aortic valve and aortic annulus, because it can both preserve a native aortic valve and dose not necessitate right ventriculotomy in resection of hypertrophied muscle and patch enlargement of interventricular septum. PMID- 11048447 TI - [Effect of Down's syndrome on perioperative and long-term prognosis after ventricular septal defect repair]. AB - In this study, we investigated perioperative and long-term prognosis and the risk of major complications after repair of ventricular septal defect in 48 patients with Down's syndrome who underwent ventricular septal defect repair between May 1980 to August 1999 were compared with those in 48 patients with normal chromosomes matched for age and time period. Pp/Ps were significantly lower after the operation in both groups; however perioperative and postoperative Pp/Ps of Down's syndrome group were significantly higher than that those of control group. The duration of intubation was significantly longer in the Down's syndrome group and the case-control study revealed that the risk of long intubation (> or = 7 days) was significantly higher in the Down's syndrome group, but the incidence of PH crisis did not differ between the 2 groups. The main reasons of prolonged intubation period were respiratory complications such as pneumonia or atelectasis. In Down's syndrome group, a 5 months old boy died of heart failure on the 5th postoperative day. All other patients were survived through a mean follow-up period of 122.4 months (the follow-up rate was 95.8%). In conclusion, the perioperative and long-term prognosis after ventricular septal defect repair in patients with Down's syndrome were similar to those in patients with normal chromosome. PMID- 11048448 TI - [The clinical application of a new method for sealing the cut surface of lung using the photopolymerized synthetic bioabsorbable hydrogel (Advaseal)]. AB - We report a case of 67-years-old male, who suffered right upper lobectomy for primary lung cancer. The upper-middle interlobar surface of right lung was incomplete interlobe. Many air leakages were noted in the cut surface of upper middle interlobe during the sealing test. We sealed this cut surface of upper middle interlobe by using the photopolymerized synthetic bioabsorbable hydrogel (Advaseal). Air leakage was not noted during the sealing test, and no leakage was not noted after operation. We have observed no biological toxicity (such as cytotoxicity, hemolytic toxicity, systemic toxicity or mutagenic toxicity) by using Advaseal. The new photopolymerized synthetic bioabsorbable hydrogel (Advaseal) is a effective sealant against pulmonary air leaks in the cut surface of the lung. PMID- 11048449 TI - [A case of ruptured distal aortic arch aneurysm with bronchiolitis obliterans organizing pneumonia]. AB - We report a case of histologically proved bronchiolitis obliterans organizing pneumonia (BOOP) associated with ruptured distal aortic arch aneurysm (DAAA) into the lung. A 63-years-old male with preoperative episode of hemosputum and hemoptysis was diagnosed DAAA. Preoperative computed tomographic scanning demonstrated that the aneurysm was surrounded with the structure of 2 layers of the enhanced high density external layer and the not enhanced low density internal layer. Combined resection of the left upper lobe and the aneurysm was performed safely because of marked adhesion between the lung and the aneurysm. Postoperative histological examination revealed that the perianeurysmal structure was due to BOOP. PMID- 11048450 TI - [A successful repair of intraoperative retrograde type A aortic dissection in a patient with aortic regurgitation and ascending aortic aneurysm]. AB - The patient was a 67-year-old male with aortic regurgitation and ascending aortic aneurysm. We noticed the type A retrograde aortic dissection occurring from the cannulation site through the right femoral artery. We discontinued cardio pulmonary bypass immediately, and established selective cerebral perfusion (SCP) eleven minutes after retrograde cerebral perfusion (RCP). We underwent simultaneous aortic valve replacement and ascending and arch graft replacement with an aid of SCP combined with RCP and systemic low flow perfusion. Postoperative course was satisfactory, although patient had a transient neurologic deficit. Intraoperative aortic dissection is a rare but potentially fatal complication. RCP may be a simple and useful method in emergency operation for intraoperative retrograde type A aortic dissection to avoid serious cerebral damage. PMID- 11048451 TI - [A case of simultaneous off-pump CABG and left lower lobectomy via left anterolateral thoracotomy]. AB - Surgical management for simultaneous pulmonary resection and cardiac surgery remains controversial. We report a case of coexisting lung carcinoma and angina pectoris who was managed successfully with a concomitant operation via the left anterolateral thoracotomy through the fourth intercostal space. After left lower lobectomy, left anterior descending and distal circumflex arteries were anastomosed with composite left internal thoracic and radial artery grafts without use of cardiopulmonary bypass. Both less invasive CABG and curative resection of lung carcinoma were achieved with these procedures. PMID- 11048452 TI - [A case of total cavopulmonary connection by utilization of coronary sinus as a hepatic venous return]. AB - A 2-year-old boy with polysplenia, double outlet right ventricle after pulmonary banding and unilateral bidirectional shunt was operated on. A modified total cavopulmonary connection was done by utilization of coronary sinus as a retrograde route for the hepatic venous return. Left SVC was transected and its distal end was anastomosed to the left pulmonary artery after PA angioplasty. An equine pericardial patch was placed over the ostia of the hepatic vein and coronary sinus. Two ostia of the coronary veins were excluded from the created route. The proximal end of the left SVC was anastomosed to the inferior side of the left pulmonary artery. Postoperative course was uneventful. The postoperative angiogram showed smooth hepatic venous return through the coronary sinus and no pressure gradient was recorded between hepatic vein and pulmonary artery. PMID- 11048454 TI - [A case of anomalous systemic arterial supply to the left basal lung with lateral chest pain and palpitation followed by left basal segmentectomy]. AB - A 38-year-old man was admitted because of left lateral chest pain and palpitation. A further examination revealed anomalous systemic arterial supply to the left basal lung. As pulmonary arteriography showed a complete lack of pulmonary arterial supply to these segments, we performed a ligation of the aberrant artery and left basal segmentectomy. Eight months after surgery, a lung perfusion scan showed improved uptake in the apical segment of the lower lobe. PMID- 11048453 TI - [A case of lung tumorlets with pulmonary carcinoid]. AB - It has been postulated that differential pathological diagnosis of lung tumorlet from pulmonary carcinoid is very important to decide the therapeutic strategy, because both of them are pathologically consisted of similar type of cells originated from same cells. At the present, although lung tumorlet is considered to be a hyperplastic lesion of Kultschitzky cells which is located in the epithelial cell of the bronchial mucosa by stimuli such as hypoxia and inflammation, but it occasionally recognized in the normal lung and the concept that it is a subtype of carcnoid is also undeniable. In this paper, a case of lung tumorlet with minute pulmonary carcinoid suggesting a subtype of carcinoid operated upon in our department is presented. Although simple pulmonary resection is a method of choice for pulmonary carcinoid, but it is still controversial as to therapeutic strategy for the lung tumorlet, because it is a benign entity and not tumorous lesion. However pulmonary resection with a close subsequent followed up study must be a best method of choice for the case with lung tumorlet includes a minute lesion of carcinoid seen in this particular case. PMID- 11048455 TI - [Croatian guidelines for management of arterial hypertension in 1999: review after one year]. AB - The guidelines for the treatment of arterial hypertension, set by the Croatian Society of Hypertension in April 1999, were largely concordant with the recommendations of JNC-VI (1997) and WHO-ISH (1999). After one year these guidelines are not only valid but even reinforced by several new lines of evidence from large prospective clinical trials. PMID- 11048456 TI - [Monitoring bacterial resistance to antibiotics in the CroatianRepublic]. AB - In 1996 a Committee for antibiotic resistance surveillance in Croatia was founded by the Croatian Academy of Medical Sciences. In this study antibiotic surveillance results for the period June 1-December 31, 1997 from 12 microbiology laboratories throughout Croatia are presented. Sensitivity to antibiotics was determined by disk diffusion method for the following bacteria: Streptococcus pneumoniae, Staphylococcus aureus, Enterococcus spp., Escherichia coli, Klebsiella spp. and Pseudomonas aeruginosa. In general, high proportion of resistant isolates was recorded throughout Croatia, although some regional variations were noticed. Mean resistance of pneumococci to penicillin was 38%, in S. aureus resistance to methicillin was 47%, and 3rd generation cephalosporin resistance in E. coli was 6% and in Klebsiella spp. 21%. In P. aeruginosa resistance to gentamicin averaged 50%, to imipenem 13% and to ceftazidim 8%. Future aims of the Committee are to continue routine antibiotic resistance surveillance during certain periods every year, and to estimate clinical significance of resistant bacteria, detect mechanisms of resistance and improve the quality of laboratory work through education and quality control projects. PMID- 11048457 TI - [Effect of the menstrual cycle and ondansetron on postoperative nausea and vomiting]. AB - The incidence of postoperative nausea and vomiting (PONV) has been studied in a prospective group of 92 female patients in the generative period, undergoing thyroid surgery. In a subgroup of 47 examinees we analyzed PONV incidence according to the surgery time within the frame of their menstrual cycle. The highest number was observed in periovulatory and premenstrual periods (p < 0.02). In the formal trial, 24 patients that randomly received ondansetron (8 mg in 4 mL i.v.) and significantly less PONV than their 21 placebo controls (4 mL saline i.v.): 4 or 16.8% vs. 11 or 53.6% (p < 0.04). PMID- 11048458 TI - [Spontaneous pneumomediastinum as a complication of asthma in adultsand adolescents]. AB - The objective of this study was to examine the percent of spontaneous pneumomediastinums (SPM) as a complication of asthma in children and adolescent, and to examine the symptoms and clinical signs which predict SPM. A retrospective analysis was performed of patients discharged from Srebrnjak Hospital between 1988 and 1991 with the diagnoses of asthma and SPM. Ten cases (8 males and 2 females) aged 4-19 years were compared with 50 hospitalized patients. SPM was found in 4.96% of patients. Mean age of patients with SPM and asthma was 11.2 and of control patients 11.45 years. Chest pain was reported in 5/10 patients with SPM and asthma and in 0/50 of controls (p < 0.01). Subcutaneous emphysema was detected in 9/10 patients with SPM and asthma and in 0/50 of controls (p < 0.01). There was no difference in respiratory distress, partial oxygen pressure and oxygen saturation, heart rate and respiratory rate between cases and controls (p > 0.05). Systolic pressure was higher in children with SPM, but the values were within normal limits. During hospitalization in 3 patients with SPM partial left pneumothorax developed, in 1 partial bilateral pneumothorax and in 2 pneumopericardium. Subcutaneous emphysema is a significant specific clinical sign, and chest pain is a predominant symptom in the diagnosis of SPM. The patients with asthma and SPM must be treated in hospital because of potential further complication. PMID- 11048459 TI - [The age and sex of hospitalized demented patients]. AB - The difference in average age and sex was examined in patients suffering from Alzheimer's dementia, vascular dementia and combined dementia. The sample comprised 454 patients discharged from the Department of Psychogeriatrics, Vrapce Psychiatric Hospital, Zagreb, during 1998. Data analysis revealed that the average age of patients with Alzheimer's dementia was 80, patients with vascular dementia 75, and patients with combined dementia 79 years. Female patients comprised 70% of the sample while there were only 30% of males. Statistical analysis of the data revealed a statistically significant difference in the average age of patients with Alzheimer's dementia in relation to those with vascular dementia, as well as differences in the average age of patients with vascular dementia and those with combined dementia. There is also a statistically significant difference in the numbers of males and females suffering from individual types of dementia. PMID- 11048460 TI - [Carcinoma in situ in the urinary bladder]. AB - In this paper we have presented seven patients with carcinoma in situ of the urinary bladder, a rare intraepithelial form of transitional cell carcinoma of the urinary bladder, described first in 1952. In all patients malignant cells were detected in urine sediment, and the diagnosis was proven histopathologicaly by random biopsies of the urinary bladder. In five patients carcinoma in situ was associated with papillary bladder tumor, while two patients had primary carcinoma in situ. We have emphasized a high rate of irritative urinary symptoms, that can lead to diagnostic mistakes. Three patients had reduced bladder capacity. In all patients a complete response was achieved after local immunotherapy or local chemotherapy. After a follow-up lasting from 23 to 61 months in one patient a recurrent carcinoma in situ was diagnosed, while six patients show no signs of recurrent disease. PMID- 11048461 TI - [BCG dissemination in a child with normal immunity]. AB - A fourteen month old girl with BCG dissemination and BCG lymphadenitis is presented. Mycobacterium bovis BCG strain was isolated from axillary fistula and from gastric contents. Pathohystological examination of the lymph mode and cytological examination of the bronchial mucus confirmed granulomatous inflammation. All tests for humoral and cellular immunity were with reference ranges. Antituberculous therapy successfully eliminated Mycobacterium bovis in gastric contents, and healed the wound in the left armpit. As the patient had tuberculoid form of BCG dissemination, the therapy was successful. PMID- 11048462 TI - [Measurement of arterial pressure with a mercury sphygmomanometer more than a technique]. AB - Arterial blood pressure has been measured using mercury sphygmomanometer and auscultatory method for more than a hundred years. The results obtained by this measurement method make the basis of almost all conclusions related to pathogenesis, epidemiology and treatment of arterial hypertension. However, some deviations from ordinarily obtained data have been observed but are undoubtedly due to superficial approach to this simple method. A number of lesser faults can together result in an eventually inaccurate conclusion both regarding diagnosis and assessment of therapeutic effects. The aim of this short review article is to remind us of those little efforts needed to be done for improving the exactness of measurement in order to increase the accuracy of results. The ultimate consequence should be better care of patients with high blood pressure. PMID- 11048463 TI - [Trachoma--an endemic and post-endemic problem]. AB - Trachoma is a specific chronic keratoconjunctivitis, characterized by follicular and papillary hyperplasia of conjunctiva, pannus, and cicatrization in the late stages of the disease. The cause of trachoma is a bacterium, Chlamydia trachomatis (serovar A, B, Ba and C). There are 146 million people in the world suffering from the active trachoma disease, and 5.9 million are blind because of it. WHO has set the goal to eliminate the blinding trachoma by the year 2020 (GET 2020 Program). The evaluation of the gravity of the disease has been made according to the Trachoma Simplified Grading System. In order to achieve the goal, it has used the SAFE strategy. The SAFE strategy comprises surgery for trichiasis, antibiotics, facial cleanliness and environmental improvement, especially in water supply and regulated sewage. The endemic trachoma in Croatia is a thing of past. Patients with active disease are rare, usually misdiagnosed and inadequately or insufficiently treated. A recent epidemic of another chlamydial (oculogenital sexually transmitted) disease has forced us to approach the diagnostics and treatment of chlamydial diseases with full responsibility. PMID- 11048464 TI - [Epidemiology of arterial hypertension in Croatia in 2000]. AB - Arterial hypertension is one of the most important modifiable risk factors for cardiovascular disease. Treatment of hypertension has been shown to decrease the risk of myocardial infarction, stroke, congestive heart failure, end-stage renal disease and peripheral vascular disease. It is apparent from some results that blood pressure should be lowered in all patients having other cardiovascular risk factors regardless whether they are hypertensive or non-hypertensive persons. However, complete information considering awareness, treatment and control of arterial hypertension is still missing. The aim of our study is to obtain these data for the entire region of the Republic of Croatia. The study will be finished in two years and new hypertension related cardiovascular risk indicators such as isolated systolic hypertension, pulse pressure and heart rate will be analysed as well. Therapeutic success will be analysed in particular subpopulations considering applied therapy. PMID- 11048465 TI - [Development of obstetrics at Knin Hospital after liberation in 1995]. AB - The objective of this study is to present the development of obstetrics in the Knin General Hospital after military action "Storm" and consecutive liberation in 1995. Delivery register was used as the source of data in the period from liberation, i.e. from August 1995 to December 31, 1998. The number of medical visits during pregnancy, number and methods of deliveries and perinatal outcome are presented. Along with the improved organization of the Obstetrics and Gynecology Department of the Knin General Hospital the number of visits during pregnancy increased from average 2.4 (0-4) in 1995 to 6.7 (2-13) in 1998. With return of the population, the number of deliveries increased from 18 in 1995 to 275 in 1998. Also, the number of Cesarean sections increased from 1 (5.6%) in 1995 to 18 (6.5%) in 1998, but the frequency is the same. Perinatal mortality was low, only one child died from twin pregnancy, as a fetal death because of cord prolapse. The results of the Department of Obstetrics and Gynecology from the Knin General Hospital are comparable with other modern centers which were not under war occupation, thanks to the great effort of the Ministry of Health and help of medical staff from Zagreb, Sisak, Sibenik, Bjelovar, Cakovec and the Knin General Hospital. PMID- 11048466 TI - [Bacterial resistance to antibiotics in Croatia in 1998 and 1999]. PMID- 11048467 TI - Evaluation of heart dysfunction using non-invasive methods. A comparison between left ventricular end diastolic volume measured using blood pool gated radioisotope angiography and left ventricular transverse end diastolic diameter measured using two dimensional echocardiography. AB - BACKGROUND: The author aimed to compare two heart function indexes by studying left ventricular end diastolic volume measured using blood pool gated radioisotope angiography, and left ventricular end diastolic diameter measured using echocardiography. METHODS: The patients were divided into two groups depending on whether left ventricular ejection fraction was greater than or equal to 50% or less than 50%, namely into a group of patients without myocardial dysfunction and one with myocardial dysfunction. The results were analysed using Fisher's discriminant analysis. RESULTS: The author observed that 41.18% of patients were correctly classified using the end diastolic volume and 82.35% using transverse end diastolic diameter. Student's "t"-test was also performed on end diastolic volumes in the first and second group, although this was not significant. On the contrary, the "t" test between the transverse end diastolic diameters of patients in the first and second group was significant with p < 0.05. CONCLUSIONS: The author concludes that the end diastolic transverse diameter of the left ventricle is a reliable index of myocardial dysfunction. PMID- 11048468 TI - Postoperative bleeding after coronary revascularization. Comparison between tranexamic acid and epsilon-aminocaproic acid. AB - BACKGROUND: Microvascular bleeding after Cardiopulmonary bypass (CPB) is mainly due to consumption of clotting factors, platelets damage, and hyperfibrinolysis. Aprotinin, the only antifibrinolytic drug effective in preserving platelets, is no longer available; an alternative regimen based on pure antifibrinolytic drugs has been proposed, since hyperfibrinolysis is known to contribute both to clot lysis and platelet dysfunction. In this study the efficacy of two antifibrinolytic drugs, Tranexamic acid (TA) and epsilon-aminocaproic acid (EACA), was tested in patients undergoing cardiopulmonary bypass (CPB), for primary myocardial revascularization. METHODS: Forty-eight consecutive patients were randomized to receive prophylactically equipotent doses of EACA (group A) or TA (Group B). Platelet count, prothrombin time, fibrin digestion products, blood loss and transfusion requirements recorded after 6 and 24 hours from the end of surgery were compared. RESULTS: The two groups were comparable for length of CPB and numbers of grafts; no significant difference was observed in the coagulation parameters considered. Blood losses were less in group B (TA) than in group A (EACA), both at 6 and 24 hours after surgery; homologous blood transfused was also less in group B, but no difference was statistically significant. No adverse effect was observed. CONCLUSIONS: In coronary patients, TA and EACA exhibit the same effects on blood loss and requirements after CPB; either drug can be safely used in cardiac surgery. PMID- 11048469 TI - Mitral prolapse. A heart anomaly in a clinical neuroendocrine context. AB - Mitral valve prolapse was identified as a separate nosological entity by Barlow in 1963. A characteristic of this cardiac anomaly is blood reflux into the left atrium during the systole owing to the lack of adhesion between valve flaps. The presence of symptoms linked to neuroendocrine dysfunctions or to the autonomic nervous system lead to the onset of the pathology known as mitral valve prolapse syndrome (MVPs). It is usually diagnosed by chance in asymptomatic patients during routine tests. MVPs includes complex alterations to the neurovegetative system and a high clinical incidence of neuropsychiatric symptoms, like anxiety and panic attacks. A neuroendocrine mechanism thought to underlie panic attacks was recently proposed based on a biological model. In general, the cardiovascular anomaly manifested by patients with MVPs could be defined in neuroendocrine constitutional terms. PMID- 11048470 TI - Physiological, metabolic, neuroendocrine and pharmacological regulation of nitric oxide in humans. AB - Genetically controlled nitric oxide production is currently assessed mainly by the colorimetric determination of the plasma levels of nitrite/nitrates, specific end-products of NO, or by the magnitude of the vasorelaxation obtained with forearm occlusion venous plethysmography after infusion of stimulating (acetylcholine) or inhibiting (N-monomethyl-arginine) substances. NO levels decrease in the elderly compared to young people and more in men than in women. Many shear stresses (physical exercise, cold pressor test, mental test) increase NO production; in these circumstances NO inhibits platelet aggregation and endothelial adhesivity: among endogenous metabolic factors mainly LDL impairs endothelium dependent vasodilation through an altered synthesis of NO, whereas triglycerides seem to have no influence; oxidized LDL may stimulate NO-production to counteract its damaging effects on endothelial cells. The hypothesis whereby an early production of NO is followed by a drop due to a prolonged cell injury is still controversial. Lipid lowering agents (drugs, apheresis) improve endothelial function. Some endogenous vasoactive substances, (acetylcholine, adenosine, bradykinin, and arginine) stimulate NO production, whereas others, such as angiotensin II, histamine, aspirin, indomethacin have inhibitory effects. As regards drugs glyceriltrinitrate and isosorbide dinitrate, acting as NO donors, improve endothelial function; other drugs are current being studied in this sense. PMID- 11048471 TI - Carotid stenosis and brain tumor. Two cases successfully treated. AB - The authors report two cases of internal carotid artery stenosis associated with brain tumors. After the presentation of the rarity of this combination little described in the literature, they point out their therapeutic behaviour explaining their technical decisions. PMID- 11048472 TI - Treatment of arterial hypertension in the elderly with diltiazem vs ramipril. AB - BACKGROUND: Around 40% of the elderly population suffer from arterial hypertension. An effective antihypertensive treatment is therefore required. Calcium antagonists are used to treat hypertension because, owing to their mechanism of action, they can provoke systemic, as well as coronary vasodilatation. In this study the authors aimed to evaluate the activity and tolerability of diltiazem compared to ramipril in a group of elderly patients suffering from essential arterial hypertension. METHODS: A controlled single blind study was performed in which patients were randomly assigned to one of two groups, A and B, consisting of 25 patients each, treated respectively with 300 mg sustained-release diltiazem or 5 mg ramipril in a single daily dose. The study lasted 6 months and evaluated systolic and diastolic pressure and heart rate. RESULTS: The evolution was positive in all patients in Group A and most patients in Group B, with the normalisation of both systolic and diastolic values. Heart rate showed a more persistent fall in Group A, but this was expected owing to the mechanism of action of diltiazem. No patient in Group A had to suspend treatment, whereas one patient in Group B had to interrupt therapy following the onset of a persistent cough. CONCLUSIONS: Both treatments resulted in similar changes in systolic and diastolic arterial blood pressure. In the light of these results, it can be affirmed that, at an oral dose of 300 mg/day, sustained-release diltiazem was found to be effective and well tolerated in the treatment of mild to moderate essential arterial hypertension in the aged. PMID- 11048473 TI - [Endometrial carcinoma: prognostic significance of cellular ploidy]. AB - BACKGROUND: The aim of the paper is to compare ploidy status with the traditional prognostic factors (grading, myometrial invasion, cytology of peritoneal fluid and node invasion) in 118 women affected by endometrial carcinoma and treated by hysterectomy in our department in order to evaluate a relationship between these parameters. METHODS: Since January 1988 and August 1996 127 women (average age: 61) affected by endometrial carcinoma and not previously treated have been submitted to abdominal or vaginal hysterectomy. A retrospective study was carried out on 118 of these women evaluating DNA ploidy on fixed neoplastic samples through flow cytometry (Coulter Elite with Argon Laser). Aneuploidy was defined as cell population containing at the same time two or more moderate peaks in G0/G1. Histology of neoplastic tissues could evaluate grading and myometrial invasion in all cases. Cytology of peritoneal fluids and nodal state were evaluated respectively in 99 and 56 patients. The results obtained have been compared by Fisher's statistical test. RESULTS: 70.3% of evaluated neoplasias were diploid, while 29.7% were aneuploid. No statistical difference was observed comparing ploidy status with every considered parameter. CONCLUSIONS: The results obtained show that DNA ploidy doesn't seem to be positively correlated with any traditional histopathological factors. The literature about this matter is questionable. Histopathological analysis is the only prognostic factor and it is the only parameter to personalize treatment. PMID- 11048476 TI - [Vulvar lesions caused by HPV]. AB - Human papillomavirus subclinical lesions are well known on the cervix and are identified by colposcopy after acetic acid staining. The transfer of this technique from the cervix to the vulva has led to the identification of areas of white epithelial changes which have been defined by similarity as vulvar subclinical HPV (VSHPV) lesions. A critical revision of the different clinical VSHPV lesions classifications, the vulvar diagnostic role of acetic acid staining, the natural history of HPV infection and the histological-biomolecular diagnostic techniques has the authors to the conclusions that the majority of the "so called" VSHPV lesions should not be considered a real disease. The presence of HPV-DNA in these subclinical lesions should be considered causal and not causal. To avoid overtreatments in women with proven HPV-DNA positivity without macroscopic clinical lesions, the authors recommend to abandon the acetic acid staining on the vulva and invite to consider the VSHPV lesions a faked diagnosis and not a clinical entity. PMID- 11048475 TI - [Celiac disease. Risk factors for women in reproductive age]. AB - In the past coeliac disease, or intolerance to gluten, has been considered a rare disease in infancy, whose most important signs were chronic diarrhea with malabsorption and reduced growth. However, besides this classical form, there are a number of other clinical and subclinical forms which may appear even in the adult life and without any overt intestinal sign. The alterations may affect, e.g., the liver, thyroid, skin and the female and male reproductive system. The overall prevalence of the different forms of coeliac disease in Western Europe is at least 1:300. The aim of the present paper is to describe and evaluate the effects of coeliac disease on female reproduction. Such effects include delayed menarche, amenorrhea, infertility and early menopause. Epidemiological studies show that besides reduced fertility, affected women are at higher risk of reproductive problems such as pregnancy loss, low birthweight of offspring and reduced duration of breastfeeding. There are no adequate studies to evidentiate a possible increase of birth defects; nevertheless, coeliac disease induces malabsorption, with deficiencies of nutritional factors essential to prenatal development such as iron, folic acid and vitamin K. The mechanisms underlying the reproductive alterations are still awaiting clarification; however, an interaction among specific nutritional deficiencies, endocrine imbalances and immune disturbances is suspected. As for the other effects associated to the coeliac disease, the possible prevention or treatment of the reproductive effects is only the lifelong maintenance of a gluten-free diet. PMID- 11048478 TI - Omphalocele and umbilical cord cyst. Prenatal diagnosis. AB - We present a case of intrauterine fetal death at 32 week's gestation with omphalocele, umbilical cord allantoic cyst and polyhydramnios. Ultrasound diagnosis of anomalies was performed at 23 weeks of gestation. Fetal karyotype was normal: 46 XX. This association has been found to have a high rate of chromosomal abnormalities, especially trisomy 18. It's difficult to explain the reason why intrauterine fetal death has happened; one possible hypothesis is that the cord cyst, compressing umbilical vessels, have caused intrauterine vascular compromise of blood flow. PMID- 11048477 TI - [Vulvar Paget's disease. Clinico-pathologic review of the literature]. AB - In 1986 the International Society For the Study of Vulvar Disease classified vulvar Paget's disease (VPD) as a non-squamous intraepithelial lesion of the vulva. The clinical multiform aspect of VPD, similar to other dermatological lesions, often delays the execution of a biopsy. Paget's cells could be instead easily identified at histological examination and with histochemical reactions. Underlying adenocarcinomas or stromal invasion are present in about 10% of intraepithelial VPD. Patients with VPD are at risk for a second synchronous or metachronous neoplasia: colo-rectal adenocarcinoma (more frequent in perianal localization of VPD), cervical adenocarcinoma, carcinoma of the transitional epithelium from the renal pelvis to urethra and mammary carcinoma. A wide spectrum of frequency of these associations is reported in the literature (0 45%). Therapy for intraepithelial VPD is wide and deep surgical resection comprising all the skin appendages. However VPD has a high frequency of recurrences (15-62%), often irrespective for radicality of surgical excision. When association with underlying invasive adenocarcinoma or stromal invasion is histologically confirmed, vulvar surgical approach must be integrated with inguino-femoral lymphadenectomy. The role of chemotherapy and radiotherapy in the multimodal approach to extensive or recurring VPD is still controversial. Recurrences or progression of intraepithelal VPD are reported more than 10 years from first surgical resection so that long term follow-up is mandatory. PMID- 11048474 TI - [Proposal of a new test for the diagnosis of PROM based on the determination of hCG in the washing fluid of the posterior vaginal fornix]. AB - BACKGROUND: The objective of this study is to determine whether or not the measurement of hCG levels in the washing fluid of the posterior vaginal fornix is useful for the diagnosis of premature rupture of membranes. METHODS: Samples were analysed from 52 normal pregnant women, divided into three groups: 20 pregnant women without rupture of membranes, 21 patients with confirmed rupture of membranes, 11 patients with suspected rupture of membranes. In order to distinguish patients of the two groups we propose a hCG cut-off value of 100 mIU/ml. RESULTS: The analysis of our data reveals that there is no overlapping between the hCG levels of the group of pregnant women without rupture of membranes and the group of patients with confirmed rupture of membranes. CONCLUSIONS: The hCG levels in the washing fluid of the posterior vaginal fornix in our experience is a useful, very cheap and non-invasive diagnostic test of PROM. PMID- 11048479 TI - [Usefulness of invasive diagnosis in an atypical case of endometrial carcinoma]. AB - Endometrial cancer generally arises early with atypical uterine bleeding: its incidence is about 8-10% in presence of this symptom. Many techniques, invasive and not invasive, have been introduced for early diagnosis and they are all more or less reliable: integration of them all is the most important objective to obtain a screening program which makes some results comparable to those obtained for cervical cancer. Hysteroscopy is the only technique which allows us to have a direct vision of the uterine cavity, of all the techniques we actually use for endometrial cancer diagnosis. Also, we can carry out hysteroscopy in the day hospital, often without anaesthesia, with little discomfort for the woman and with little risk of complications: these facts speak well for this technique. We discuss a case of a 71-year-old patient, para 1001, menarche at 13, menopause at 45, affected by a G2-G3 endometrial adenocarcinoma located in an atypical site, for which diagnosis hysteroscopy was of primary importance. PMID- 11048480 TI - Biotechnology and meaning. PMID- 11048481 TI - [Too late for Paul Kammerer]. PMID- 11048484 TI - Phenotypic plasticity in hydrozoans: morph reversibility. AB - The life cycle of Hydrozoans typically comprises two phases: the polyp, either solitary or colonial, with generally a benthic habitat, and the medusa which lives in the plankton. In its typical metagenetic cycle, the medusa is budded from the polyp, which is the product of sexual reproduction of medusae. However, several alternative reproduction patterns have also been described. In particular some species are able to perform a regressive transformation of the medusae that transform themselves into polyps bypassing sexual reproduction. In a species with alternative morphs switched by the environment, the more labile is the correlation between environmental factors acting on the genetic switch and the factors to which the resulting form is adapted, the more hazardous will be the development of either body form. However, we can explain the evolutionary advantage offered by reversion between morphs of these plastic species living in shallow water unpredictable environments: should produced medusae be released in the "wrong" environment, they would still have a chance of survival under another form. PMID- 11048482 TI - The "sense of being stared at" does not depend on known sensory clues. AB - The "sense of being stared at" can be investigated by means of simple experiments in which subjects and lookers work in pairs, with the looker sitting behind the subject. In a random sequence of trials, the looker either looks at the back of the subject, or looks away and thinks of something else. More than 15,000 trials have already been conducted, involving more than 700 subjects, with an extremely significant excess of correct over incorrect guesses (Sheldrake [1999]). This effect was still apparent in experiments in which subjects were blindfolded and given no feedback, showing it did not depend on visual clues, nor on the subjects knowing if their guesses were right or wrong (Sheldrake [2000]). In this paper I describe experiments I conducted in schools in England in which the subjects were not only blindfolded and given no feedback, but looked at through closed windows. There was again a very significant excess of correct over incorrect guesses (p < 0.004). At my request, schoolteachers in Canada, Germany and the United States carried out similar experiments and found an even more significant positive effect than in my own experiments (p < 0.0002). The fact that positive results were still obtained when visual clues had been effectively eliminated by blindfolds, and auditory and olfactory clues by closed windows, implies that the sense of being stared at does not depend on the known senses. I conclude that peoples' ability to know when they are being looked at depends on an influence at present unknown to science. PMID- 11048487 TI - Not only fibroblasts but also melanoma cells express "leucine aminopeptidase" activity. AB - "Leucine aminopeptidase" (LAP, aminopolypeptidase, EC 3.4.11) activity has been recommended and widely used as a histochemical marker for the identification of contaminating LAP-positive fibroblasts in pigment cell cultures. Using a sensitive biochemical assay with L-leucyl-p-nitroanilide as a substrate we demonstrated that in vitro melanoma cells also exhibit LAP activity. Our comparison of four melanoma cell lines with four fibroblast lines showed that the differences in the enzyme activity were not qualitative but only quantitative. For this reason the specific antibodies, karyological analysis and electron microscopy are recommended as more reliable means in distinguishing fibroblasts from poorly differentiated pigment cells than the LAP-cytochemistry. PMID- 11048488 TI - Differences in the NADPH-diaphorase positivity of the cholinergic brain stem neurons following damage of their thalamic termination field. AB - Following electrolesions of the anteroventral thalamic nucleus (the target structure for the NADPH-diaphorase positive axons of the cholinergic mesopontine cells, e.g. laterodorsal, pedunculopontine and lateral parabrachial nucleus) the intensity of NADPH-diaphorase staining changed drastically, but unlikely in each nucleus: it increased in the cells of the laterodorsal nucleus on the side of the lesion and decreased on the contralateral nucleus, whereas the staining intensity of the pedunculopontine and parabrachial cells remained unchanged bilaterally. In cases with small (partial) lesions of the anteroventral thalamic nucleus only lower number of heavily stained cells were observed in the laterodorsal nucleus ipsilateral to the thalamic lesion, whereas the number of slightly stained cells in the contralateral laterodorsal nucleus decreased. In contrast, not even large lesions of other thalamic nuclei, anterior or tuberal hypothalamus, zona incerta, pallidum internum, striatum or corpus callosum changed the symmetric intensity of the NADPH-diaphorase staining of the laterodorsal, pedunculopontine or lateral parabrachial nuclei. Increased intensity of the NADPH-diaphorase staining in the cells of the mamillary body was found two weeks after the electrolesions of the ipsilateral mamillo-thalamic tract or anteromedial thalamic nucleus. No degenerative changes or decrease of number of neurons in the laterodorsal tegmental nucleus in Nissl staining were detected after the electrolesions. Our data show that electrolytic lesions of target areas can lead to an upregulation of NOS expression in the parent cell bodies, provided that there is no wide collateralization as found for the pedunculopontine or parabrachial nuclei. PMID- 11048483 TI - The discovery of the chemical nature of tobacco mosaic virus. AB - The path to arrive at the elucidation of the chemical nature of plant viruses was greatly facilitated by the availability of Tobacco mosaic virus (TMV) as biological tool. The first hypothesis on the chemical nature of TMV was advanced in 1899 by the American Albert Wood, who suggested an enzyme nature. This hypothesis, severely questioned by Harry Hallard in 1915, was re-proposed by several virologists. In 1926, the American Maurice Mulvania concluded that the virus might be a protein with the biological characteristics of an autocatalytic enzyme. Before arriving at the experimental evidence it was necessary to resolve two questions: the estimation of virus infectivity in quantitative terms, performed by Francis Holmes in 1928, and the purification of the virus, performed by Carl George Vinson between 1927 and 1934. Vinson gave a conclusive contribution to solve the question of the chemical nature of TMV by settling the protocol of TMV purification. He put forward the hypothesis of the protein nature in the early 1930s but had not the required firm belief to gave the final experimental evidence of it. Who first arrived at the experimental evidence of the protein nature of the virus was the American Wendell Meredith Stanley, in 1935. His celebrated work, a classic of the fundamental Virology, was followed by several papers in which this result was firmly reaffirmed. The heuristic value of Stanley's discovery held out a year: the decisive evidence of the actual chemical nature of TMV was offered in the late 1936 by an English group under the leadership of Frederick Charles Bawden. In their short paper, Bawden and co operators demonstrated that TMV had a ribonucleoprotein nature, a result that was confirmed in the following years for several TMV strains and other viruses. Stanley and his group did accept this result only after a year of reticence and contradictions. The conversion to the ribonucleoprotein nature raised a dignified protest by Bawden and the sarcasm of his closest co-operator, Norman Wingate Pirie, because Stanley proved to be very reluctant to recognize the merit of the English group. The world of Virology continues to consider Stanley as the first scientist who elucidated the actual nature of a virus, and this eminent scientist was awarded the Nobel Prize for Chemistry, in 1946. By examining the papers Stanley published from 1937 to 1945, one can however find proof of his ambiguity, a fact that justifies the bitterness of Bawden and the sarcastic comments of Pirie. PMID- 11048485 TI - The emergence of emergence: a critique of "design, observation, surprise!". AB - Artificial life research begins from the premise that Alife subsumes real life. A criterion for emergence in Alife has been formulated that, however, excludes real life and postulates the need for a real life Designer and an Observer. This in effect nullifies the premise of Alife and takes us back to the argument for God from design of Bishop Paley in 1802. An alternative is to realize that Alife could include two properties: simulated organisms that both design themselves and are the observers. Self-design can come about via evolution in a population of mating organisms, especially via mutations that are gene or higher order duplications. Duplications permit novelty while retaining previously attained functions. The ability to observe can itself evolve, if its construction process evolves. This may now be possible to simulate, if new paradigms for embryogenesis, such as positional information or differentiation waves, prove accurate, or at least sufficiently robust to construct a wide diversity of observational abilities. The evolution of perception, however, may be limited by the physics available to the Alife organisms, which can come in three forms: simulated physics, real physics accessible to robots, or "Cyberspace physics". PMID- 11048489 TI - Development of multiple organ failure in critically ill newborns under AT III substitution. AB - Criteria for the AT III substitution were fulfilled by 33 infants. Mean birth weight was 1321 g (480 g up to 2860 g), and mean gestational age was 29 weeks (23 39 weeks). AT III substitution in dosing of 5 u/kg/hr was administered to all patients as well as standard heparin 10 u/kg/hr. Initial plasma level of AT III in the study group prior to start of the substitution was 45 +/- 17% activity of normal plasma of adult donor. The value increased up to 90 +/- 32% after 24 hours and up to 113 +/- 38% at the end of the therapy. Eight patients died (24%). Considerable decrease of the Neonatal Therapeutic Intervention Scoring System (NTISS), indicating a level of multiple organ dysfunction, was observed as early as after the first 48 hours of AT III therapy in the entire group (p < 0.01). Extensive increase of platelet count (p < 0.05) occurred after 48 hours of AT III therapy in survivors. Except for a severe sepsis in one case, no clinical signs of increased bleeding occurred during the AT III therapy, and the clinical signs of pre-existing bleeding even disappeared in 5 cases. Analysis of the group of 8 deceased patients revealed irreversible organ dysfunction in 7 cases present prior to the start of AT III substitution. In conclusion, during the AT III substitution, no bleeding, no clinical side effects but improvement in multiple organ dysfunction including increase in platelet count was recorded. PMID- 11048486 TI - [Adolf Portmann, Wilhelm Troll: two biologists of idealistic morphology]. PMID- 11048490 TI - Development of fixable stent for long-term derivation of lower urinary tract. AB - The object of the envisaged project is the development of easily fixable non irritant stent of the lower urinary tract. The project has the aim to remove the obstruction and make passage-way through urethra which, after repeated operations, remains narrowed and requires repeated dilatations or, in the patients who have a chronic epicystostomy due to the non-passable urethra. The prosthesis must be self fixable, non-irritant, easily introduceable and removable by endoscopy, enabling normal urination with urinary continence and, last but not least, at a reasonable price. PMID- 11048493 TI - [Josef Hynie (1900-1989)--100 years since the birth of the founder of Czechoslovak sexology]. AB - The anniversary of one hundred years from the birth of Josef Hynie, the renowned professor of Charles' University and pioneer of medical sexology, is an impulse for thinking again about his lifelong successful creative activities, namely clinical, scientific, publication, teaching and sexual-educational activities. At the Faculty of Medicine in Prague Hynie enforced and built the first university sexological institute in the world and achieved its general recognition. With the team of his pupils for years he integrated the behavioral and reproduction aspects of the branch and thus predetermined its wide conception also for the future generations of Czech sexologists. In 1981 Hynie's efforts were crowned by definitive inclusion of medical sexology into the Czechoslovak health care system of that time. The authors pay attention also to his unforgettable, excellent character traits. The biographical treatise is framed up in an authentic remembrance of MUDr. Jaroslava Pondelickova-Maslova, his colleague for many years, and enlivened by photographs of that period. PMID- 11048492 TI - [The personality of psychotherapists] . AB - The profession of the psychotherapist is very demanding and his or her personality, constituting one of the effective factors of the treatment, affects the entire psychotherapeutic process. The psychotherapist's desired personality traits include inner stability and high degree of self-knowledge leading to an understanding and accepting of his or her own self. The other useful personality traits--the therapist's openness for the patient as well as his or her unconditional acceptance, empathy and authenticity--create an optimal and safe therapeutical setting enabling the patient to communicate freely and sincerely. All these traits must be expressed simultaneously, otherwise they could be destructive. They are underscored by the therapist's ethical conduct towards the patient. Ignoring these moral principals anulls any positive influence of the above traits. The conclusion makes mention of Kant's categorical imperative. PMID- 11048491 TI - The speech intelligibility at the opera singing. AB - The authors investigated the speech intelligibility at opera singing. They analysed several arias sung by soprano voices from Czech opera "Rusalka" (The water Nymph), from Puccini's opera "La Boheme" and several parts of arias from "Il barbiere di Siviglia" sung by baritone, tenor, bass and soprano. The sonographic pictures of selected arias were compared with a subjective evaluation. The difference between both authors was about 70%. The opinion is, that the singer's formant is not the only one problem having its role in the speech intelligibility of opera singing. The important role is played by the ability to change the shape of vocal tract and the ability of rapid and exact articulatory movements. This ability influences the shape of transients that are important at the normal speaking and also in the singing. PMID- 11048494 TI - [Specific features of the clinical course and angiographic pattern in chronic coronary artery occlusion]. AB - Particular features of coronary angiography and clinical presentation of coronary artery disease have been studied in patients with chronic total coronary occlusion. Chronic total coronary occlusion is defined as TIMI 0 or TIMI I type flow in the artery for more than three days. Patients with coronary occlusion have more severe course of coronary artery disease: they more often suffer myocardial infarction and high gradations of angina. Myocardial function is much more affected if there is occlusion of left descending artery, or there are no signs of intercoronary collaterals. PMID- 11048495 TI - [Immediate and long-term outcomes of the use of a Crossflex wire coronary stent in the treatment of patients with different forms of ischemic heart disease]. AB - Immediate and medium-term prognoses were studied in patients with coronary heart disease in whom a Crossflex wire coronary stent (Cordis, Johnson & Johnson, USA) had been inserted into the coronary arteries. The study indicated that it could be effectively used in patients with both chronic coronary heart disease and its exacerbation (acute myocardial infarction). It was found that in the vast majority of patients anginal attacks ceased and the clinical course ran smoothly in both immediate and medium-term postoperative periods. In the immediate period, various complications were observed only in 6.7% of cases. In the late period, the incidence of restenosis was 24%. If restenosis occurred, repeated balloon dilatation was successfully made in the great majority of cases by resulting in recovery of the vascular lumen. PMID- 11048496 TI - [Thoracic computed tomography after surgical interventions on the lung]. AB - The paper based on the analysis of computed tomographic data in 40 patients again underlines the importance of accurate and timely diagnosis of thoracic changes in the postoperative period. It shows that CT can provide information on thoracic organs and parts and presents the tomographic pattern of a postoperative uncomplicated period. PMID- 11048498 TI - [Significance of the method of fractal dimension assessment of x-ray images in the differential diagnosis of spherical formations of the lungs]. AB - A posterior processing of tomograms was made in 210 patients with spherical formations of the lungs of various etiological factors. The method based on tomograms delicate texture study according to the system of 7 diagnostic signs by means of the fractal dimension of X-ray pictures of the pathological focuses made the tomography diagnosis more informative in lung spherical formations substrate recognition. The method of fractal estimation allowed to reach a 14.7% rise of the precision, a 12.6% rise of the sensitiveness and a 17.1% rise of the specificity of diagnosis as compared with the routine X-ray research. PMID- 11048497 TI - [Comparative evaluation of the diagnostic efficiency of interactive digital radiography and routine fluorography in screening examinations of the chest]. AB - The current trend for decreased exposure to medical ionizing radiation sources contributes to the introduction of novel technologies into radiation diagnosis. The aim of this study was to compare the informative value of interactive digital X-ray study and routine enlargement fluorography. The study covered 316 individual without the complaints typical of lung diseases. Three roentgenologists assessed X-ray images by 7 major signs. The sensitivity, specificity, and accuracy of the methods in question were defined. The final stage was the construction of routine operating curves (ROC). When there were diffuse changes in the lung, the sensitivity of interactive digital X-ray study was 14% higher. The higher informative value of interactive digital X-ray study is also attested by the higher position of a respective ROC. The fact that lung tissue function in combination with low radiation load may be estimated assumes the use of interactive digital X-ray study in patients with pneumosclerosis and emphysema. PMID- 11048499 TI - [Current comprehensive diagnosis of pancreatobiliary cancer complicated by jaundice syndrome]. AB - The study was undertaken to enhance the efficiency of diagnosis of pancreatobiliary cancer complicated by jaundice by using the potentialities of currently available instrumental studies at most. The data of examination of 256 patients with this disease are presented. Based on diagnostic findings, the authors have constructed a diagnostic model of precision recognition of cancer of the extrahepatic bile ducts, pancreatic head, Vater's papilla. PMID- 11048500 TI - [Radiographic diagnosis of injuries of the pelvic ring in acute trauma]. AB - Data on 58 victims with multifocal damages to the pelvic ring were used to examine the diagnostic potentialities of different radiation diagnostic techniques and to compare their resolving power. The latter was 65.1, 83.3, and 94.7% in plain and multidimensional X-ray studies, and computed tomography, respectively. The paper describes the complex of signs of closed sacral fractures on plain X-ray films and oblique pelvic inlet (cauda), proposed by the authors, which could improve the diagnosis of fractures by 8.8 times, and an original orthopedic gauze-plate for the detection and estimation of invisible pelvic bone displacement, and an original procedure for pelvic X-ray study with targeted load in acute injury. This all can improve the quality of examination of the injured substantially and define indications for different treatments more correctly. PMID- 11048501 TI - [Magnetic resonance tomography in the study of the thyroid gland]. PMID- 11048504 TI - [Intravascular ultrasonography in the year 2000]. PMID- 11048503 TI - [CT in the diagnosis of gastric cancer (review of the literature)]. PMID- 11048506 TI - [Effect of antioxidant therapy on indicators of free radical activity in workers at risk of lead exposure]. AB - The authors administered to 36 workers exposed to lead, for four weeks the preparation Prolong which contains vitamin C, vitamin E, beta-carotene, selenium, zinc and chromium and investigated the effect on indicators of free radicals. A statistically significant increase of the following occurred: superoxide dismutase (p < 0.001), glutathione peroxidase (p < 0.01), an increase of the total antioxidant capacity (p < 0.01) and an increase of uricaemia (p < 0.05). Based on these results the authors consider supplementation with antioxidants indicated in workers exposed to lead. PMID- 11048505 TI - [Haematopoietic stem cell mobilization in patients with multiple myeloma- comparison of 3 variations of the stimulating regime]. AB - In the literature various regimes are described for successful collection of haematopoietic stem cells in patients with multiple myeloma. Most frequently cyclophosphamide is used, 5 g/m2 combined with different doses of haematopoietic growth factors. In our group the yields and tolerance of three stimulating regimes are compared: 1. cyclophosphamide 5 g/m2 and filgrastim (G-CSF) 5 micrograms/kg 2. cyclophosphamide 5 mg/m2 and filgrastim 10 micrograms/kg 3. cyclophosphamide 5 g/m2 and figrastim 5 micrograms/kg along with erythropoitin 50 IU/kg. Cyclophsphamide is administered always on the first day and the haematopoietic growth factors then from the third day till the end of collection of haematopoitic cells. In patients with multiple myeloma where only four cycles of VAD chemotherapy preceded and where radiotherapy of the axial skeleton was not used, optimal collection of haematopoietic stem cells was achieved after administration of cyclohosphamide 5 g/m2 with subsequent administration of 5 micrograms/kg G-CSF. By increasing the dose of G-CSF to 10 micrograms/kg or adding 50 IU/kg erthropoietin did not lead to a significant improvement of the tolerance and yield of this procedure. PMID- 11048507 TI - [Can we determine which coronary artery is affected by the localization of ST depression? Comparison of ergometric and coronarographic results]. AB - The authors evaluated retrospectively 206 patients with ischaemic heart disease. These patients were subjected to an ergometric loading test on a bicycle ergometer followed by angiography of the coronary arteries. With regard to the site of the ST depressions during the loading test the patients were divided into 7 groups which were further subdivided according to the coronographic finding. The authors investigated the hypothesis that patients with depressions of the ST segment above the anterior wall suffer from an affection of the ramus interventricularis anterior, patients with depressions of ST above the lateral wall from affection of the ramus circumflexus and patients with depressions above the lower wall have an affection of the right coronary artery. From the work ensues that no site of the ST depression correlates significantly with the coronary finding. The correlation is not statistically significant even when we compare the site of ST depressions with the affection of one artery regardless of the affection of the other coronaries. PMID- 11048508 TI - [Early rehospitalization in cardiovascular diseases]. AB - The authors investigated perspectively during 1998 the frequency of early rehospitalizations (within 30 days after discharge) focused on cardiovascular diseases. From a total number of hospitalized patients (2349) early rehospitalized patients accounted for 91% (214 rehospitalizations). Rehospitalizations after cardiac attacks accounted for 45.4% rehospitalizations and for 4.1% of all hospitalized patients. The largest ratio of rehospitalizations on account of cardiovascular diseases were patients with heart failure (39.1%), with cardiac arrhythmias (22.6%) and unstable angina pectoris (21.6%). In relation to the number of total hospital admissions most frequently patients with unstable angina pectoris were rehospitalized within 30 days after discharge (20.1%). Least frequently patients with myocardial infarction, a cerebrovascular attack and hypertension were rehospitalized (3.1%, 3.0%, 1.9%). An important factor which was followed up was the cause of rehospitalization which was assessed in 54.6% of patients. In patients with heart failure in as many as 71%. The authors assume that in this percentage of rehospitalizations it should be possible to avert it and to reduce thus at the same time economic costs. It was not possible to assess exactly the role of social factors in rehospitalizations but it is possible that they were identical with the cause on the part of the patient and participated by 17.5-21% in rehospitalizations (see tables 7 and 11). PMID- 11048502 TI - [X-ray endoscopic methods of external and internal drainage of the bile ducts in mechanical jaundice]. PMID- 11048515 TI - [The concept of internal medicine as a profession]. PMID- 11048509 TI - [Pedal bypass in the treatment of critical ischemia in the diabetic foot]. AB - A formerly established theory on obliteration of diabetic foot arteries was dismissed. In the last decade, diabetic foot revascularization using so called very distal bypass has became a routine procedure. The reconstructions are undertaken in patients with chronic critical limb ischemia. This paper presents our initial experience with this, not yet widely used, operative technique in 16 patients who had a total of 14 pedal bypass procedures performed. The parameters we monitored included the 3-, 6-, and 12-month patency rates, defect healing, and limb salvage rates. The primary and secondary patency rates were 64.3% and 78.6% at 3 months; 55.5% and 66.7% at 6 months; and 50% and 75% at one year, respectively. The overall limb salvage rate for the above periods was 92.9%. All defects healed following successful revascularization using pedal bypass. In patients where bypass could not be established, limb salvage was accomplished in one in three cases only. PMID- 11048510 TI - [Changes in bone mineral density in Th12 to L5 vertebrae in female patients with osteoporosis]. AB - The authors examined 47 female patients, age 50-60 years, with primary and secondary osteopenia and osteoporosis. In each patients they assessed the bone mineral density (BMD) of the cortical and trabecular bone of vertebrae Th12 to L5 by quantitative computer tonmography (QTC). They did not include in the group patients with deformities of the vertebrae grade one, two and three. These deformities were assessed from a lateral X-ray of the thoracic and lumbar spine. OBJECTIVE: To evaluate the importance of assessed BMD values of vertebrae from the biomechanical aspect. STATISTICAL ANALYSIS: Individual groups of assessment of BMD in cortical and trabecular bone for each vertebra were characterized by means of 95% confidence intervals and their means or median values. RESULTS: The BMD of cortical bone increases from Th12 to L5 almost in a linear fashion. BMD of the trabecular bone declines from Th12 to L3, in L4 abd L5 it rises again. The lowest values were recorded in L3. The greatest decline of BMD of the trabecular bone by 24.05% in L3 as compared with reference values of the given age groups were not conditioned by deformities of the vertebrae. The BMD of cortical bone in osteopenic and osteoporotic patients practically did not differ from reference values for the given age group. CONCLUSION: From the presented work ensues that deformities of the vertebrae do not occur as long as the BMD of cortical bone is within the range of reference values despite a decline of BMD in trabecular bone. It is thus more important to investigate the material qualities of cortical bone because the risk of fractures of the vertebrae depends on the elasticity and firmness. Critical BMD values of cortical bone assessed by QCT for the development of fractures are not known so far. QTC of vertebrae due to its higher radiation load and financial costs is not a routine densitometric method. It will be necessary to develop new densitometric methods which will make it possible to assess BMD or another property of cortical bone and assess critical values for the development of vertebral fractures. PMID- 11048512 TI - [Intravascular ultrasound in cardiology]. AB - Intravascular ultrasound (IVUS) is a clinically useful tool that provides cross sectional images of the coronary arterial lumen and wall. Diagnostic applications of IVUS include the evaluation of ambiguous lesions on angiography particularly at the bifurcations. IVUS is also useful in the assessment of coronary vasculopathy in cardiac transplant patients or it can help to diagnose abnormalities such as syndrome X or coronary artery spasm. IVUS can optimize the performing of percutaneous coronary interventions, especially stent implantation. It represents as well an optimal tool for assessing regression of atherosclerosis. Three-dimensional reconstruction, elastography and imaging guide wires are some of the recent advances in the field of intravascular ultrasound. PMID- 11048514 TI - [Intracoronary ultrasound: the "platinum standard" in evaluating the morphology of coronary stenoses. Does it have a role in clinical practice?]. AB - The authors present the case of a 45-year-old man with a history of myocardial infarction of the anterior wall and unstable angina pectoris where they detected a critical stenosis of the proximal segment of the ramus, interverticularis anterior by intracoronary ultrasound examination. The subsequent coronary angioplasty with implantation of a stent was also implemented under ultrasound control. The authors discuss the contemporary position of intravascular ultrasound in coronary interventions in particular with regard to the development of restenosis. PMID- 11048516 TI - [Dual endopeptidase inhibitors--a new direction in the development of hypertensive agents]. AB - Endogenous peptidases participate in a major way in the formation of peptide pressor substances such as angiotensin II (A II) and endothelin (ET) as well as in the degradation of depressor substances, e.g. atrial natriuretic peptide (ANP) or bradykinin. They include on the one hand the angiotensin converting enzyme (ACE) and endothelin converting enzyme (ECE), on the other hand kinase II for bradykinin and neutral endopeptidase 24.11 (NEP) for ANP. Inhibition of these enzymatic reactions leads to a decline of vasopressors A II and ET and conversely delays the break-down of vasodilatating bradykinin and ANP. The main haemodynamic consequence of this double inhibition is a reduced peripheral vascular resistance and decline of the blood pressure. The concurrent block of both systems (dual inhibition) is more effective than the isolated block of one substance. The first dual endopeptidase inhibitors were ACE inhibitors blocking the conversion of angiotensin I to A II and inhibiting at the same time the degradation of bradykinin by kininase II which is identical with ACE. At present further substances were synthetized with a dual inhibitory effect e.g. on ECE and on NEP (phosphoramidone, thiorphan, ecadatril etc.). Under experimental conditions they have a long-term antihypertensive effect on the vascular wall and heart muscle. The development of another dual ACE and NEP inhibitor has reached already the stage of clinical tests and the first clinical studies. The preparation omapatrilate in amounts of 2.5-80 mg significantly reduced the BP in a dose dependent way in mild and medium advanced essential hypertension. Normalization of the blood pressure, i.e. a drop below 140/90 mm Hg, was achieved with omapatrilate monotherapy in as many as 83% of patients with hypertension stage I and in 53% patients with essential hypertension stage II. The drop of blood pressure after 20-80 mg/day depended on the degree of hypertension and was comparable or better than monotherapy with lisonopril 20 mg/day or amlodipine 10 mg/day. Treatment with omapatrilate was well tolerated. Dual peptidase inhibitors interfering with the formation of pressor substances and with the degradation of depressor substances seem to be a perspective class of antihypertensives also useful in the treatment of other cardiovascular diseases (heart failure, primary pulmonary hypertension). Before its final inclusion in the therapeutic pattern, further comparative and clinical mortality studies must be implemented. PMID- 11048511 TI - [Monitoring minimal residual disease in pediatric patients with acute lymphoblastic leukemia]. AB - The level of minimal residual disease is an important prognostic factor in childhood acute lymphoblastic leukaemia. The end of induction therapy is the most significant time-point for prediction of treatment outcome. Within a pilot study covered by the Paediatric Haematology Working Group in the Czech Republic 51 childhood patients were analysed at diagnosis of acute lymphoblastic leukaemia and at the end of induction using method based on detection of clonal rearrangements of immuno-receptor genes. The majority of tested patients (32/51, 63%) had a low or non-detectable levels of residual disease, a group of patients with the highest levels and thus the highest risk of relapse included 10% of patients (5/51). Within each of three risk groups one patient has relapsed so far. Therefore, the relapse rate in particular subgroups is 3% (1/32), 7% (1/14) and 20% (1/5) to date, respectively. The results are compared with these published by the BFM group (van Dongen et al., Lancet 1998). The pilot phase of a new BFM treatment protocols includes examination of residual disease for stratification of patients into the different risk groups. PMID- 11048513 TI - [Triacylglycerol as a risk factor in early ischemic heart disease]. AB - Triacyglycerol levels above 2.0 mmol/l are according to contemporary findings an independent risk factor for the development of ischaemic heart disease. The triacylglycerol concentration of plasma provides information on the amount of lipoproteins, which carry also a major amount of cholesterol and which can also accelerate the process of atherosclerosis. Hypertriglyceridaemia as an independent risk factor was confirmed in particular in large epidemiological studies--the Framingham and Munster study. An increased triacyglycerol concentration implies obviously a greater risk for women than for men. From published results of intervention studies ensues moreover that reduction of triacyglycerol levels during hypolipidaemic treatment may reduce the incidence of ischaemic heart disease, coronary attacks and lead to regression of coronary atherosclerosis. In patients with hyperlipoproteinaemia therapeutic efforts should therefore focus not only on reduction of LDL-cholesterol but also on correction of hypetriglyceridaemia. PMID- 11048518 TI - [Ciprofibrate in the treatment of combined hyperlipoproteinemia. Results in more than 600 patients from 23 centers in the Czech Republic]. AB - A total of 633 patients with combined hyperlipoproteinaemia from 23 centres in the Czech Republic met the criteria for assessment of the effectiveness of 3 month administration of 100 mg ciprofibrate (Lipanor) per day. The cholesterol concentration declined from the original values of 6.94 mmol/l by 13%. The drop of triglycerides was even more significant from 3.03 mmol/l by more than 41%. The relatively low HDL cholesterol at the beginning, 1.14 mmol/l, increased by 15%, while LDL-cholesterol dropped significantly by almost 12%. The classical atherogenic index expressed by the ratio of total/HDL cholesterol declined by 25%. Treatment was very well tolerated by the patients and was associated with a minimum of undesirable side-effects. Treatment with ciprofibrate is effective and safe treatment in patients with combined hyperlipoproteinaemia. PMID- 11048519 TI - [Beta-blockers in the treatment of chronic heart failure. How should results of clinical studies be introduced into clinical practice]. AB - The long-term action of elevated catecholamine concentrations on the heart muscle causes a number of adverse effects from apoptosis to malignant arrhythmia or terminal failure of the cardiac pump. Recently completed mortality studies with beta-blockers (BB) in chronic heart failure CIBIS II and MERIT HF confirmed the conclusions of pilot studies with carvedilol that in stabilized chronic heart failure BB reduces the mortality rate as well as the need of hospital admission on account of deteriorating heart failure. Proper selection of patients for administration of BB is necessary, assessment of absolute and relative contraindications. It is also necessary to define the initiation and increase of dosage and the target dose of the beta-blocker. The sooner treatment with beta blockers is started, the greater the foreseen benefit for the patient. PMID- 11048517 TI - [The new atherogenic plasma index reflects the triglyceride and HDL-cholesterol ratio, the lipoprotein particle size and the cholesterol esterification rate: changes during lipanor therapy]. AB - The new atherogenic plasma index (AIP) is a logarithmic transformation of the ratio of the molar triglyceride (TG) concentration and high density lipoprotein cholesterol (HDL-C). AIP correlates closely with the size of LDL particles (r = 0.8) and esterification rate of plasma cholesterol devoid of apo B lipoproteins (FERHDL), r = 0.9 which are considered at present the most sensitive indicators of the atherogenic plasma profile. AIP was recommended by the authors, based on analysis of results of 11 previous studies (1156 subjects) where FERHDL and plasma lipid parameters were investigated in different groups of people who differed as to the atherogenic risk. The AIP index was moreover used for evaluation of a clinical study comprising 609 patients with hyperlipidaemia, who were treated for three months with ciprofibrate (Lipanor). The mean AIP values of non-risk groups (plasma from umbilical blood, children, healthy women etc.) equalled zero or were lower, while with an increasing atherogenic risk (men, women after the menopause) AIP reached positive values, incl. high positive values in risk groups (plasma of diabetic subjects, patients with HLP, patients with positive angiography, myocardial infarction etc.). In all groups women had lower AIP values as compared with males. In patients after Lipanor therapy the AIP declined (from 0.58 +/- 0.17 to 0.33_0.18 in men, from 0.50 +/- 0.18 to 0.21 +/- 0.19 in women). If we consider AIP values from negative ones to 0.15 as "safe" from the aspect of atherogenicity, before Lipanor treatment these "safe" levels were recorded in 1.5% men and in 5.2% women and after treatment in 32% men and 48% women. The results indicate, that AIP which reflects the plasma lipoprotein profile quantifies the relations between TG and HDL-C and thus can be an objective indicator of the atherogenic risk and effectiveness of treatment and it is useful because it can be assessed in any surgery. PMID- 11048520 TI - [Angiotensin I converting enzyme: what results from new findings] . AB - Angiotensin I converting enzyme (ACE) participates not only in the regulation of the extracellular fluid volume and blood pressure but also in the control of proliferating processes in the human organism. The favourable effect of ACE inhibitors in the treatment of essential hypertension may be caused also by their antiproliferative effect. The development of essential hypertension is probably of polygenic origin. The gene for ACE, and its alleles encountered commonly in the population is involved in the pathophysiology of systemic hypertension by acting on the dynamic properties of processes in the organism which regulate changes of the extracellular fluid volume and proliferation. Therefore it is necessary, when treating systemic hypertension with ACE inhibitors, to pay attention to monitoring of the blood pressure and organ changes. The relationship with blood pressure assessed by conventional methods need not be close. The antiproliferative effect of ACE inhibitors may be favourable in the treatment of hypertension and its clinical use in oncology may be taken into consideration. PMID- 11048521 TI - [IgG subclasses and autoantibodies in adult patients with selective IgA deficiency]. AB - Selective IgA deficiency is the most common primary immunodeficiency disease, but the etiopathogenesis is unknown. In a portion of patients disturbed IgA production is accompanied by various immunological abnormalities. Serological laboratory results of 30 female and 22 male adult patients with selective IgA deficiency were compared with sex- and age-matched healthy controls. Hypergammaglobulinemia IgG was observed in 31 patients and only in 2 control persons. Serum IgG1 was increased in 12 and/or IgG3 in 18 patients, the increase in IgG2 was less common (6 persons). The number of persons with increased IgE did not differ from the control group. The occurrence of autoantibodies (antinuclear antibodies, rheumatoid factor, antithyroglobulin and anti-thyroid microsomal antibodies, anti-gastric parietal cells, reticulin, smooth muscle, anticardiolipin, and anti-gliadin antibodies) did not differ significantly from the control group. IgG hypergammaglobulinemia, which, according to our results, is the most frequent accompanying serological abnormality in IgA deficiency, may be caused by compensatory increased production, but may also reflect more profound immunological dysregulation in the disease. PMID- 11048522 TI - [Chronic heart failure in the aged]. AB - Ageing of the population, advances in the treatment of cardiovascular diseases and in the treatment of chronic heart failure cause a rapid increase of the prevalence of chronic heart failure in the population and of the number of hospital admissions on account of this diagnosis. The majority of patients with chronic heart failure are over 65 years and their ratio is increasing. The diagnosis clinical course and treatment of old patients with chronic hearth failure has specific features which become more marked with advancing age. As a role other associated diseases are present as well as factor which cause deterioration of chronic hearth failure, complicate the diagnosis and treatment. With regard to the large number in the population and the patients age ambulatory care of patients with chronic heart failure will also in future be mainly ensured by general practitioners in close collaboration with cardiologist and specialist in internal medicine. A further progression of the number and severity of hospital admissions on account of chronic heart failure may be foreseen. PMID- 11048523 TI - [Cardiotoxicity of antineoplastic therapy]. AB - Cardiotoxicity is a serious complication of anti-tumorous treatment. Cytostatics can cause a number of undesirable side-effects such as arrhythmias, angina pectoris, acute myocardial infarction, sudden death, cardiac failure. The probably most serious cardiotoxicity is chronic cardiac failure after treatment with anthracyclines. Interest in the diagnosis, monitoring and treatment of cardiotoxicity revealed new findings of cardiac complications after various cytostatics, high-dosage chemotherapy and transplantation of haematopoietic cells. Prospective paediatric studies provided evidence of the serious character of late cardiotoxicity of anthracyclines. The authors review the most frequent cardiac complications of anti-tumorous treatment. They emphasize in particular the toxicity of anthracyclines and its possible prevention. PMID- 11048524 TI - [Principles of conservative treatment in chronic pancreatitis]. AB - The authors summarize the principles of conservative treatment of chronic pancreatitis with special attention to treatment of exocrine pancreatic insufficiency and pancreatic pain. They emphasize the principles of the rational administration of drugs with pancreatin, the position of analgesics, and other possible procedures, if medicamentous treatment fails. An outline of dietetic treatment is also given. PMID- 11048525 TI - [Geriatrization of internal medicine]. AB - The author draws attention to the need of more profound knowledge of geriatrics in the training of specialists in internal medicine. He draws attention to the ageing population, trends in advanced countries and planned enlistment of general specialists in internal medicine into primary care. PMID- 11048527 TI - [The prehospital phase in patients with acute myocardial infarct in Slovakia. A challenge]. AB - Better management of patients with acute myocardial infarction during the prehospital phase is at present a challenge not only for health workers but for society as a whole. The authors pay attention to knowledge of the complex problem of the prehospital phase in patients with acute myocardial infarction which is a prerequisite for finding possible solutions for a favourable effect on their management. The authors analyzed 3,040 patients who were admitted to hospital alive within 96 hours after the development of complaints with suspicion of a first or repeated acute myocardial infarction. They focused attention in particular on prehospital time delay. They found that within a satisfactory time interval (within 2 hours) 29.8% patients were delivered and within a yet acceptable interval of 4 hours 51.6% patients (with respect to effectiveness of thrombolytic treatment). Similarly undesirable are also data on the patient time delay. Within the optimal first hour after development of complaints following the decision of the patient (subjects present) to ask for or seek medical assistance was the decision of 34% patients, during the first two hours 47.5% and within 4 hours 61.2% patients with acute myocardial infarction. The ratio of time delay of the patient in the total prehospital delay is 45.5% even in patients who were admitted during the first hour after development of acute myocardial infarction. In patients who were admitted 4 hours after development of complaints it is 79.5%. The patient is admitted to hospital most quickly if he calls the medical emergency service and latest when he decides to see a doctor. Physicians and other health workers contributed only in 16.4-20.9% patients with acute myocardial infarction to their early decision to seek medical assistance. Patients with an early decision (within one hour) call most frequently the medical emergency service and are taken to hospital by this service. The time delay due to transport is shortest in these patients. The late hospital admission of patients with acute myocardial infarction in Slovakia calls for reduction of the time interval from the development of complaints to hospital admission (total prehospital delay), in particular the time taken by the patient to make up his mind (patient time delay). Early calling of the emergency medical service and transport of the maximum possible number of patients with acute myocardial infarction to hospital by the emergency medical service will greatly improve the management of patients with acute myocardial infarction. Comprehensive implementation of the survival chain (24), the most comprehensive implementation of recommendations of the European Society of Cardiology and the European Resuscitation Council for in the management of patients with acute heart attacks during the prehospital phase (21) and early effective treatment which begins already in the prehospital phase has a favourable impact on the condition of patients with acute myocardial infarction and on their prognosis. Along with early and effective treatment of patients with acute myocardial infarction in hospital and their stratification these are the most important approaches to the development and control of sudden cardiac death. Knowledge of the complex problem of the prehospital phase in patients with acute myocardial infarction is the prerequisite for the elaboration of high standard prehospital management of patients with acute myocardial infarction. PMID- 11048528 TI - [Ventricular fibrillation in chronic heart disease]. AB - The objective of the work was to describe in subjects with spontaneous ventricular fibrillation, after elimination of acute cardiac disease, the strategy of antiarrhythmic treatment and to evaluate, based on prospective follow up, the effectiveness of this treatment. The authors included in the group 36 patients (30 men and 6 women) within the range from 34 to 78 years (mean age 58 +/- 11 years) with spontaneous ventricular fibrillation. They divided the group into a subgroup (15 subjects) without revascularization of the heart muscle, into a subgroup (17 subjects) with revascularization of the myocardium (coronary angioplasty and bypasses) and a subgroup (4 subjects) where ischaemic heart disease was ruled out (mostly cardiomyopathies). In all subgroups they used programmed ventricular stimulation (apparatuses of Quinton Co. USA, Biotronik Co. GFR), in the subgroup with revascularization within 3 months. During the diagnostic procedure of ventricular stimulation they tested antiarrhythmic drugs most frequently amiodarone per os (for 4 weeks). An implantable cardioverter- defibrillator was implanted in 17 patients (8 subjects without revascularization, 6 subjects with revascularization, 3 subjects without ischaemic heart disease). All patients were followed up till death, maximum 24 months. The authors evaluated the rate of cardiac deaths (death on cardiac grounds, incl. sudden arrhythmic death) and sudden arrhythmic deaths (within one hour after the onset of symptoms or the first malignant ventricular tachyarrhythmia recorded after implantation of the defibrillator). In the subgroup without revascularization with electric instability of the ventricles according to programmed stimulation 66.7% they described seven cardiac deaths (46.7%) and 6 sudden "arrhythmic" deaths (40%) incl. 5 subjects with ineffective testing of antiarrhythmic drugs. Conversely in the subgroup with revascularization and with diagnostic programmed stimulation in 47.1% they found 3 cardiac deaths (17.7%), one sudden "arrhythmic" death (5.9%)--a subject with ineffective testing. In the subgroup without ischaemic heart disease they recorded cardiac and sudden "arrhythmic" deaths in half the subjects, in all instances in subjects without inducible ventricular tachyarrhythmia. The authors found in the course of a two-year investigation a relapse of cardiac arrest in 25% of subjects after spontaneous ventricular fibrillation. A third of these subjects (all without a cardioverter defibrillator) died. They confirm the benefit of implantation of a defibrillator for all subjects regardless of the basic diagnosis and revascularization of the heart muscle. PMID- 11048529 TI - [Effect of bisoprolol on heart rate variability in patients with hyperthyroidism]. AB - OBJECTIVE: The objective of the submitted work was to assess by the method of spectral analysis of the heart rate how the use of bisoprolol will affect the autonomous modulation of the heart rate in hyperthyroid patients and what are the time relations of this effect. MATERIAL AND METHODS: In the investigation 10 female patients with thyreotoxicosis caused by Graves-Basedow's disease were included. Their average age was 38 years and along with a thyrostatic drug bisoprolol was administered. Spectral analysis of the heart rate was made before the onset of treatment, after a week and after six weeks with the patients in an upright and recumbent position. The activity of the sympathicus was evaluated from the ratio of low and high frequencies. RESULTS: Already during the first week a significant decline of the heart rate was observed. During the first week of treatment a decline of the activity of the sympathicus in a recumbent position occurred associated with milder palpitations or their complete disappearance. The decline in an upright position occurred after the first week of treatment. From the trend of other parameters of cardiac variability, the differences of some reached statistical significance, it may be concluded that the activity of the parasympathicus, formerly inhibited, was restored. CONCLUSION: Based on the results it may be stated that bisoprolol is a suitable supplementary drug in the early treatment of thyrotoxicosis because by reducing the hyperadrenergic state it improves the quality of life of these patients. PMID- 11048531 TI - [Adult patients after surgery of ostium primum type of atrial septal defects in childhood: echocardiography study]. AB - The authors examined, using transthoracic and transoesophageal echocardiography, 36 adult patients (15 men) aged 22 +/- 3.1 years (18-29 years) who were operated 12.2 +/- 3.7 years previously on account of a defect of the atrial septum type ostium primum. In these patients no other congenital cardiac defect was present. In addition to closure of the defect in the patients complete suture of the "cleft" of the anterior cusp of the mitral valve was performed, in 7 partial suture of the "cleft" of the anterior cusp of the mitral valve and in 4 commissuroplasty. In one instance later reoperation with replacement of the mitral valve by a mechanical prosthesis was performed. The control group was formed by 16 healthy volunteers (5 men) aged 22.1 +/- 3 years (19-31 years). Patients operated in childhood on account of an atrial defect of the ostium primum type have on echocardiographic examination, as compared with healthy volunteers, larger atria and the left ventricle, a thicker interventricular septum and left ventricular wall and a higher velocity of left ventricular filling during the late diastole. Higher values of parameters of the size and volume of the left ventricle are associated with the presence of mitral regurgitation. More marked changes of systolic or diastolic left ventricular function are not present, there are not even any echocardiographic signs of higher pressure in the atria and pulmonary artery. In none of the patients a residual shunt at the level of the atrial septum is present. Mitral regurgitation is found in two thirds of the patients, only in one case it was however moderately severe (grade 3). From the results it does not ensue which type of surgery of the "cleft" of the mitral valve has the best long-term results. In none of the patients tricuspid regurgitation of a higher grade than grade 1 is present. PMID- 11048534 TI - [Plasma pH and anticoagulant solutions used in the plasmapheresis method of blood collection from donors]. AB - The resulting pH of fresh frozen plasma for clinical use, collected by plasmapheresis from blood donors is influenced by the type of anticoagulant solution and its ratio with the donor's blood. The authors describe the use of three anticoagulant solutions with a different sodium citrate concentration and different ratios of donor blood. As compared with the physiological range of pH of the blood, the resulting pH value of the collected plasma, when using ACD-A and AB-16 solutions, varies within the range classified as acidosis, i.e. less than 7.36. When using a 4% sodium citrate solution the plasma pH value is in the area evaluated as alkalosis. The authors discuss indications for administration of fresh frozen plasma in clinically serious diseases and the influence of administration of this transfusion preparation on the acid-base balance as the transfusion recipients are threatened by the development of metabolic acidosis. Maintaining the pH value of fresh frozen plasma slightly above the physiological range of blood pH prevents in particular during massive plasma transfusions the possibility of deterioration of acidosis or its development. PMID- 11048530 TI - [Radiofrequency catheter ablation of left-sided atrioventricular accessory pathways. Immediate and long-term results using the trans-aortic approach]. AB - Catheter mapping and ablation of left accessory pathways may be difficult if the pathway fibre orientation is oblique across the mitral annulus. This is particularly true in case of concealed accessory pathway that conducts only in ventriculoatrial direction. The purpose of the study was to compare catheter ablation safety, efficacy, procedure and fluoroscopy times and number of radiofrequency energy deliveries for manifest and concealed left accessory pathways using retrograde transaortic approach. 56 patients (26 females) aged 41.6 +/- 15.1 years underwent 59 ablation procedures for 59 left accessory pathways. 25 (44.6%) patients with 27 concealed accessory pathways and additional 2 patients with a manifest pathway mappable only during retrograde conduction formed group I and were compared to 29 patients with a manifest accessory pathway mappable during ventricular preexcitation (group II). Ablation was acutely successful in all patients. Reablation was necessary for the pathway recurrence in 2 patients and for a different pathway in 1 patient. Deep femoral venous thrombosis complicated ablation in 1 patient. During the 14.9 +/- 8.2 (3-30) month follow-up period all the patients remain free of a pathway related tachyarrhythmia. Both the groups (I vs II) did not differ in procedure (180 +/- 76.7 vs. 187 +/- 75.8; p = 0.36) and fluoroscopy (25.3 +/- 18.6 vs. 33.5 +/- 24.7; p = 0.08) times (min) and number of radiofrequency energy deliveries (10.0 +/- 9.2 vs. 10.2 +/- 9.8; p = 0.46). CONCLUSION: Transaortic approach to the left concealed accessory pathways is safe and effective in virtually 100% of patients and, compared to manifest pathways, does not prolong procedure and fluoroscopy times nor the number of radiofrequency energy deliveries. PMID- 11048535 TI - [Postural trauma and rhabdomyolosis causing acute renal failure]. AB - Rhabdomyolysis (damage of the muscles of various origin) leads to the efflux of the intracellular fluids in the circulation. The common complication of this status is the renal failure. The early diagnosis and the proper treatment makes the fall of renal function reversible. That is why the possibility of the rhabdomyolysis must be consider. The case report describes the development of renal failure in young, previously healthy men, followed by trauma mechanism after drug and alcohol abuse. PMID- 11048532 TI - [Evaluation of kidney function in patients before and after orthotopic liver transplantation]. AB - Subjects before and after orthotopic transplantation of the liver develop frequently renal disorders. Evaluation of the renal function based on the serum creatinine concentration or its clearance is difficult in these cases because the concurrent hyperbilirubinaemia usually interferes with the method used for creatinine assessment (based on Jaffe's positive chromogen) and gives incorrect lower values. In the submitted work the authors investigated the development of the serum creatinine concentration and serum bilirubin concentration in 50 subjects before and one year after transplantation of the liver. The findings suggest that although under conditions of hyperbilirubinaemia creatinine cannot be evaluated by the usual methods, it is possible to obtain clinically useful information by investigating the trend of the relationship of creatinine and bilirubin. A decline of bilirubin is associated with a rise of creatinine which need not imply a decline of renal function, as long as creatinine does not exceed the upper range of normal values. Conversely the rise of bilirubin associated with a drop of creatinine need not imply improved renal function. In the investigated group one year after transplantation of the liver an impaired renal function (creatinine > 105 mumol/l) at a normal bilirubin level (< 21 mumol/l) in 17 (49%) subjects was found. In 7 (64%) subjects with hyperbilirubinaemia the creatinine value was within a normal range--this finding however does not rule out reduced renal function. Evaluation of renal function with concurrent hyperbilirubinaemia calls for more accurate methods. With regard to the mentioned findings however investigation of the trend creatinine/bilirubin makes it possible to evaluate the dynamics of changes of renal function. PMID- 11048533 TI - [Latent membrane protein LMP-1 in Hodgkin's lymphoma]. AB - The authors examined using the antibody against latent membrane protein (LMP-1) a group of 169 patients with Hodgkin's lymphoma (age 2 to 82 years). From the total number of 169 patients 48 (28%) patients were positive when tested with this antibody. In the whole group the following histological types were represented most frequently: type II (nodular sclerosis) 83 (49%) patients, and type III (mixed cellularity) 70 (41%) patients. Type I (lymphocytic predominance) was not represented. In type IV (lymphocyte depletion) there were three cases (1.7%). Type V (lymphocyte rich) was represented by 13 patients (7.6%). The frequency of positive cases was in these histological types as follows: type II 14 cases (17% of 83 cases), type III 28 cases (40% of 70 cases), type IV 2 cases (66% of 3 cases). type V 4 cases (30% of 13 cases). Distribution of positive cases by age: in children under 10 years a positive finding was recorded in 80%. In old people above the age of 80 years there was a 100% positivity (only two patients were examined). The smallest number of positive cases was in the third decade (of 26 patients 4% were positive). LMP-1 positivity was most frequent in male patients- in 37 (of 96 examined patients) and in 11 female patients (of 73 examined) The frequency of LMP-1 in Hodgkin's lymphoma is consistent with similar studies in economically developed countries. A markedly higher incidence of positive cases in the lowest and highest age groups and gender differences are striking and so far there is no unequivocal explanation for them. PMID- 11048536 TI - [Thrombotic thrombocytopenic purpura--still a therapeutic problem. Case report of a patient on long-term therapy]. AB - Thrombotic thromboctopenic purpura is a rare multisystemic life threatening disease the treatment of which is still a serious problem. The most successful therapy is plasmapheresis where the whole plasma volume of the patient is replaced by fresh frozen plasma or cryosupernatant. The authors describe a 34 year-old patient with the chronic relapsing form of the disease and repeated cerebrovascular attacks with an ischaemic genesis with developed organic psychosyndrome. The only effective treatment of the patient are exchange plasmaphereses replacing plasma by cryosupernatant after the patient became refractory to fresh frozen plasma. The authors describe the clinical development of the disease, its treatment by plasmaphereses and mention various ways of a venous approach and associated problems. Ensuring a venous approach in patients with this disease is of vital importance and in the terminal stage of the disease it is extremely difficult. A solution, though temporary, is implantation of a vascular prosthesis, Diastat. The functioning of the implant in this disease is however greatly threatened by the development of thrombotic occlusions. PMID- 11048537 TI - [Pacemaker dysfunction during use of a mobile telephone]. AB - Ubiquitious electronic equipment increases the number of potential sources of electromagnetic interference with sufficient energy to influence a pacemaker function. Besides others the mobile phone play important role. A 76 year-old female patient with pacemaker was hospitalized because of bilateral congestive heart failure. During ECG telemetry a short episode of pacemaker dysfunction was recorded. In a period of 15 seconds at first a pulse output inhibition occurred, then an undersensing was present. In all the 24-hour recording no other similar episode was observed. Such an utterly atypical picture lead to a direct question on mobile telephone calling. The patient confirmed that during a visit of her relatives she was really using a mobile phone. The GSM (Global System for Mobile Communication) phones operate on a carrier frequency of 900 MHz which is modulated generally in pulses about 8 Hz and 2 Hz. These signals can be falsely detected as an intrinsic heart activity by a pacemaker or they are analysed to be an electromagnetic interference. As a result a pacemaker inhibition, an asynchronous pacing and in dual chamber systems also a ventricular triggering can be present. These reactions usually do not occur if the distance between phone and pacemaker is higher than 10 cm. With some measures patients with pacemakers can safely use mobile phones. In pacemaker dependent patients a careful approach is necessary. PMID- 11048538 TI - [Imidazole receptor agonists--a new advance in the treatment of hypertension?]. AB - Agonists of I1 imidazolin receptors are a new drug groups which was registered for the treatment of hypertension. Their antihypertensive action is comparable with current antihypertensives (hydrochlorothiazide, enalapril, atenolol, nifedipine retard) and causes a drop of the systolic BP by cca 15-20 mm Hg and a drop of the diastolic BP by 10-15 mm Hg with a probable normalization of the blood pressure in cca 60% patients with mild to moderate hypertension. Agonists of I1 imidazoline receptors are suitable in particular for the treatment of hypertension associated with metabolic syndrome. Their effect in patients with ischaemic heart disease or after a cerebrovascular attack is not known and despite very promising theoretical prerequisites they are not indicated in patients with chronic heart failure. PMID- 11048539 TI - [Diagnostic approaches in pleural effusion]. AB - It is not easy to discover the underlying cause for pleural effusion. Over a quarter of the pleural effusions remain unexplained by indirect methods of examination such as chemical, cytological or bacterial tests. Thoracoscopy however, shows impressive results and can be conducted under simple conditions. Different statistical collections allow for the assumption that pleural effusions due to cardiac disease and malignant conditions occur equally often. They are followed in frequency by infection diseases and the often misjudged effusion with pulmonary embolism. PMID- 11048542 TI - [Preliminary reflections on radiation exposure from diagnostic X-rays]. PMID- 11048540 TI - [High resolution electrocardiography of P-wave signals in clinical cardiology]. AB - Signal-averaged electrocardiography of the QRS complex detects presence of late potentials which represent significant arrhythmogenic marker responsible for increased risk of life-threatening ventricular arrhythmias especially in post myocardial infarction patients. The P wave signal-averaged electrocardiography has been designed to predict development of atrial fibrillation in different populations of patients. Many P wave signal-averaging methodological procedures have been developed. However their common disadvantages remain lack of standardization, relatively low specificity and sensitivity, low positive predictive value as well as limited number of larger prospective clinical trials. Recent software and technological improvements of the P wave high resolution techniques as well as some new reports about the influence of antiarrhythmic drugs on the parameters of the P wave signal-averaged electrocardiography are creating new possibilities not only for the diagnosis of paroxysmal atrial fibrillation but also for the prediction of antiarrhythmic drug efficiency. PMID- 11048541 TI - [Teleradiology]. PMID- 11048543 TI - [Exposure and good practice in helical computed tomography]. AB - Helical CT is the imaging modality that delivers the highest exposure to patients. For each acquisition, the average radiation dose is between 20 and 30 mGy. This dose can potentially be reduced by decreasing the intensity or voltage and by increasing the pitch. Helical data can be reformatted to obtain images with smaller increment or multiplanar reconstructions, hence reducing the need for additional acquisitions. Hardware and software devices designed for dose reduction must be systematically used. Operators of CT units should be aware of the radiation dose delivered with helical CT and must carefully assess the need of each additional acquisition. Obsolete protocols such as angulation of the gantry for lumbar CT, which increases patient exposure, should no longer be used. Rational use of helical CT decreases radiation exposure and is faster, whereas improper use increases radiation exposure without added benefit. PMID- 11048544 TI - [MRI imaging in cortical laminar necrosis]. AB - PURPOSE: The goal of this study was to follow over time the MR imaging features of cortical laminar necrosis. PATIENTS AND METHODS: Six patients with cortical laminar necrosis were included. There were two women and four men aged 54-84 years, with a mean age of 68 years. In four patients, cortical laminar necrosis was caused by ischemic stroke, one case occurred after a cardiac arrest and the last patient had a meningoencephalitis. The time delay from insult to the first MR study varied between one week and 3 months. RESULTS: The MRI showed hyperintense lesions in the cerebral cortex on T1W and T2W images. The high intensity signal was still observed a few months after the insult. Cortical laminar necrosis lesions did not demonstrate hemorrhage on CT and MRI studies. CONCLUSION: MRI allowed detection of cortical laminar necrosis and could differentiate it from hemorrhage. PMID- 11048545 TI - [Evaluation of the efficacy of foraminal infusions of corticosteroids guided by computed tomography in the treatment of radicular pain by foraminal injection]. AB - PURPOSE: To evaluate the efficacy of foraminal steroid injections performed under CT guidance for the management of radicular pain. METHODS: Periganglionic infiltrations were performed in 160 patients with radicular pain refractory to medical treatment. Imaging showed either degenerative foraminal stenosis, herniated disk or postsurgical fibrosis. RESULTS: 102 patients (63.8%) had significant pain reduction. Pain relief was lasting in 68 (66.6%). CT showed the position of the needle tip, as well as the diffusion of the therapeutic compounds. CONCLUSION: We consider that CT-guided periganglionic steroid injections should be an integral part of the management strategy for radicular pain resistant to medical treatment. PMID- 11048546 TI - [Radio-anatomic study of the angle of origin of dysplastic renal arteries]. AB - PURPOSE: To compare the angulation at the origin of dysplastic renal arteries compared to atherosclerotic renal arteries, in order to improve the technique for percutaneous transluminal angioplasty of FMD. MATERIALS AND METHODS: Retrospective analysis of 40 aortograms in patients who underwent renal angioplasty for dysplastic stenosis, with comparison with 45 control aortograms (with or without atherosclerotic lesions of the renal arteries). The angle of implantation of the renal arteries was measured relative to the aortic axis in the frontal plane, taking into account only the angulation of its proximal segment. We identified three types of kidneys with regard to their position relative to the renal artery ostium. RESULTS: The angle of implantation of dysplastic renal arteries is significantly sharper compared with the control group (63.8 degrees vs 80.9 degrees, p = 0.0001), irrespective of the side. The angulation did not correlate with the position of the kidney or the direction of the renal artery, suggesting a congenital origin. CONCLUSION: The angle of implantation of the dysplastic renal arteries relative to the aortic axis in the frontal plane is sharper than the angle measured in non dysplastic renal arteries. PMID- 11048547 TI - [Preoperative evaluation of translabyrinthine cholesteatomas by MRI]. AB - PURPOSE: Hearing preservation is one of the major goals of surgical resection of invasive cholesteatomas. Patients were prospectively evaluated using a 3D-CISS MR acquisition in order to improve the detection of perilymphatic fistulae. MATERIALS AND METHODS: 16 patients (10 M, 6 F) presenting with a primary (1 case) or a secondary (post-otitis) middle ear cholesteatoma extending to the osseous labyrinth (as defined by HR-CT) were evaluated at MR (1.5 T Vision, Siemens). The 3D-CISS sequence (TR: 12.25 ms, TE: 5.90 ms, flip angle: 70 degrees, slice thickness: 1 mm, matrix: 307 x 512, FOV: 200) allowed detection of invasion of the membranous labyrinth using a 3 level grading scale: 1) normal fluids, 2) focal and 3) diffuse obliteration of labyrinthine fluid. RESULTS: While CT showed a definite osseous labyrinthine fistula, the 3D-CISS sequence depicted either a normal membranous labyrinth (9 cases), a focal obliteration of the basal turn (1 case) or lateral semi-circular canal (3 cases) or a diffuse obliteration of the labyrinthine fluid (3 cases). Diffuse obliteration of the labyrinth fluid and 1 out 3 cases of focal obliteration of the semi-circular canal were found to have perilymphatic fistulae at surgery. None of the normal labyrinths were associated with perilymphatic fistula formation. CONCLUSION: The 3D-CISS sequence allows a comprehensive preoperative evaluation of the membranous labyrinth. Loss of signal from the labyrinthine fluid due to invasion or compression of the membranous structures accounts for the lack of specificity of the technique. PMID- 11048548 TI - [Late solitary metastasis of a renal cancer to the contralateral adrenal gland]. AB - We report a case of metachronous metastasis from renal cell carcinoma to the contralateral adrenal gland detected one year after radical nephrectomy. The initial tumor was incidentally discovered in the setting of acute aortic dissection. A large left adrenal tumor was detected on CT follow-up at two years. Retrospectively, a hypervascular lesion was present on the first yearly CT examination. Adrenalectomy was performed. There is no evidence of recurrent disease at 12 months. The patient was also treated with oral steroids. Because of their location and of the particularities of available therapeutic options, and because these metastases can occur late, long-term sonographic and CT follow-up should be performed. The clinical, imaging, therapeutic and prognostic aspects of these lesions will be discussed. PMID- 11048549 TI - [Ectopic pancreas located in the small bowel: CT-endoscopic appearance. Apropos of two cases]. AB - Two patients have been diagnosed preoperatively with ectopic pancreas located within the small bowel using CT-enteroclysis. This technique, combining helical CT and small bowel opacification through a naso-jejunal tube, allows detection of small tumors. PMID- 11048550 TI - [Arteriovenous fistula following femoral angioplasty]. AB - We describe a case of arteriovenous fistula developing at the site of balloon dilatation immediately after percutaneous transluminal angioplasty of the superficial femoral artery. This occurred during an otherwise uncomplicated angioplasty. The fistula was treated by inflating a slightly larger balloon with a good clinical result as confirmed by color Doppler ultrasound. This uncommon complication of angioplasty is not widely recognized. PMID- 11048553 TI - [Optimization of a teleradiology system between two geriatric hospitals and a radiology department of a CHU]. AB - The routine use of a 64 kb/sec Numeris network requires that a compromise be reached between image quality and time required for image transfer. Our goal was to present our experience and describe our choices when reporting examinations for two geriatric hospitals located at a short distance from our radiology department. Different digitization parameters were tested (resolution, compression) for 10 plain films corresponding to the usual clinical situations: abdominal X ray, chest X ray, wrist, spine and pelvis X rays. A subjective analysis of the global quality and of the quality of a region of interest enlarged twelve times has been performed. After result analysis, our choices were: digitization using 75 or 150 DPI and systematic use of a reversible compression format. This system proved satisfactory for both clinicians and radiologists; since its implementation, 1,800 files have been reviewed. PMID- 11048552 TI - [What is your diagnosis? Unusual dilatation of the Virchow-Robin space]. PMID- 11048551 TI - [Atypical forms of Mayer-Rokitansky-Kuster-Hauser syndrome: role of echocardiography]. AB - We present three cases with atypical presentation of the Mayer-Rokitansky-Kuster Hauser syndrome observed by US between August 1994 and December 1997. In two cases we observed complete absence of the uterine. In the last case, an uterus remnant was present. In all cases, both ovaries were normal and the three women reported "normal" sexual activity. An ectopic kidney was present in one patient with complete absence of the uterus. Atypical forms of Mayer-Rokitansky-Kuster Hauser syndrome with normal vagina can be diagnosed at US. In the cases presented here, US allowed a clear description of the anatomy and provided a comprehensive evaluation of the patients. PMID- 11048554 TI - [Summary of the SFR seminar "Teleradiography"]. PMID- 11048556 TI - [Effect of anti-osteoporosis agents on the incidence of vertebral fractures]. AB - In the early nineties of the last century, increase in bone mineral density (BMD) was usually the primary endpoint in studies observing the effect of anti osteoporotic medication. Recently, several large studies have been published with the number of new vertebral deformities as the primary endpoint. This should be preferred, since fractures are generally associated with clinical signs and symptoms, while changes in BMD are not. In all these studies of the effects of alendronate, risedronate, raloxiphen and calcitonin in postmenopausal women with osteoporosis there was a striking discrepancy between a small increase in BMD of the lumbar spine, varying from +2 tot +8%, and a much larger reduction in the number of patients with new vertebral fractures: -36 to -49%. This difference could be related to an effect on bone quality, an independent effect on bone resorption, or to technical limitations of BMD measurements that underestimate true effects of antiresorptive therapy. The number of patients needing to be treated to prevent one fracture depended on background risk and on the effectiveness of the therapy. PMID- 11048555 TI - [Influenza A pneumonia]. AB - The majority of influenza cases are not associated with complications. Secondary bacterial pneumonia, commonly caused by Streptococcus pneumoniae or Staphylococcus aureus, is well known to most clinicians. Primary influenza viral pneumonia, characterized by rapidly progressive hypoxia and respiratory insufficiency together with non-consolidating pulmonary infiltrates, has a high mortality rate. In 3 patients, a man aged 74 years, and two neonates aged 11 months and 4 weeks respectively, primary influenza A pneumonia was diagnosed. In the latter two patients the virus was cultivated from sputum. Despite intensive supporting and drug treatment, the first and the last patients died. In view of evolving therapeutic possibilities, notably regarding neuraminidase inhibitors, it is important that clinicians recognize this complication of influenza at an early stage. PMID- 11048559 TI - [Diagnostic image (6) (Pustulous skin lesions)]. AB - During chemotherapy a 35-year-old woman developed pustulous skin lesions. Biopsies of these lesions showed Candida albicans. Blood cultures were positive for C. albicans. The patient died. PMID- 11048558 TI - [Roaming through methodology. XXV. Outcome measures, surrogate outcomes, and intermediate measures]. AB - Clinical medicine is aimed at decreasing the current or future burden of disease. Disease outcome is best expressed as burden of disease or change thereof, and as such the only true 'outcome measure' in clinical research. However, outcome research is expensive in costs and time expenditure, and sometimes impossible. Therefore study results are often expressed in intermediate measures. These measures tell us something about the disease process and the pathophysiological consequences of the disease and should have a relation with outcome. If this relation is strong, the measure is called 'surrogate outcome'. Intermediate measures and surrogate outcomes have advantages and disadvantages. The reader of clinical trial results first has to decide whether the answer to the study question is relevant in his personal situation. If so, the applicability of a measure can be simply appraised by answering 3 questions: 'Is the measure truthful (relevant, unbiased)?'; 'Does it discriminate between situations that are of interest?'; 'Is the measure feasible in my setting?' PMID- 11048560 TI - [Calculation of costs of stroke, cost effectiveness of stroke units and secondary prevention in patients after a stroke, as recommended by revised CBO practice guideline 'Stroke']. AB - OBJECTIVE: Economic analyses have been part of the revision of the Dutch multi disciplinary stroke guidelines. We evaluated the recommendations on stroke units and prevention of stroke recurrencies in terms of medical costs and health effects among stroke patients. DESIGN: Cost calculation. METHOD: Mathematical modelling of medical costs per patient and costs per life year gained without severe stroke (Rankin score (> 3)), by age and sex for each guideline. RESULTS: Lifetime costs of stroke depended on age and sex and vary between 84,000 and 292,000 Dutch guilders (HFL). The cost-effectiveness of stroke units decreases with age and varies between HFL 37,000 and HFL 60,200 with a large uncertainty range. Four of seven options in secondary prevention were cost-effective by previously established criteria (< HFL 40,000 per year gained without severe disease). Acetylsalicylic acid remained the drug of choice for monotherapy with dipyridamol as a second choice in patients without atrial fibrillation. Clopidogrel was not cost-effective at the current cost level. Anticoagulation after stroke in case of atrial fibrillation was cost-effective. CONCLUSIONS: Given a short hospital stay stroke units can be as affective as other hospital interventions. Acetylsalicylic acid is the most cost effective monotherapy for secondary prevention. PMID- 11048557 TI - [Treatment of severely delayed gastric emptying]. AB - For the drug treatment of gastroparesis, domperidone, metoclopramide and cisapride may be prescribed as prokinetics. Positive effects on the rate of emptying of the stomach, dyspeptic symptoms and quality of life are best documented for cisapride. Simultaneous use of cisapride with substances that inhibit the metabolism of cisapride or that may lengthen the QT interval, is contraindicated because of the risk of arrhythmias. Erythromycin is a powerful prokinetic, but because of its antibiotic effects it is usually prescribed only for a brief period. For patients who in spite of drug treatment have persistent unacceptable symptoms and keep losing weight, invasive treatment should be considered. The first step is then insertion of a jejunal tube, followed, if necessary, by antrectomy with Billroth-I reconstruction. The next step is subtotal gastrectomy with Roux-Y reconstruction, this may result in abatement of the symptoms, which, however, rarely disappear altogether. PMID- 11048564 TI - Changes in volumetric bone mineral density after gastrectomy as assessed by dual energy X-ray absorptiometry. AB - We used dual energy X-ray absorptiometry (DXA) to study changes in estimated volumetric bone mineral density (EstVBMD) of the lumbar spine after gastrectomy. The study group comprised 41 men and 32 women. When EstVBMD was compared according to sex among patients younger than 60 years of age, patients 60 to 69 years of age, and patients these three groups in men (0.185 g/cm3, 0.187 g/cm3, 0.187 g/cm3, respectively). In contrast, EstVBMD was significantly lower in women 60 to 69 years of age (0.157 g/cm3) and those 70 years of age or older (0.159 g/cm3) than in women younger than 60 years (0.200 g/cm3) (P < 0.01). When the relation between EstVBMD and the number of months after gastrectomy was studied according to sex in patients younger than 70 years, EstVBMD negatively correlated with the interval after operation in men (r = -0.365, P < 0.05), whereas there was no correlation between these variables in women. These results suggest that after gastrectomy bone mineral density decreases gradually in men younger than 70 years, but not in women. The lack of a consistent change in bone mineral density after gastrectomy in women is apparently caused by the marked effect on bone metabolism of decreased female hormone levels after menopause. PMID- 11048562 TI - [Influenza season 1999/2000 and vaccine composition for the season 2000/01]. AB - The first signs of influenza activity in the Netherlands during the 1999/2000 influenza season were the isolation of an influenza A (H3N2) virus in week 40 and of two more in week 43 of 1999. From week 50 onwards, a strong increase of the clinical influenza activity was observed which reached its peak in weeks 1 and 2 of 2000 and then rapidly declined. The clinical influenza activity was associated with the isolation of predominantly influenza A (H3N2) viruses. Near the end of the epidemic, influenza A (H1N1) and influenza B viruses were isolated sporadically. The antigenic properties of the influenza A (H3N2) viruses resembled those of the epidemic strains isolated in the previous season and the vaccine strain A/Sydney/5/97. This influenza season, influenza B viruses did not play a significant role and they matched the vaccine strain B/Yamanashi/166/98. In addition, a small number of influenza A (H1N1) viruses were isolated. Some of these viruses resembled the old variant of influenza A (H1N1) viruses, A/Bayern/7/95, whilst others showed a close antigenic relationship with the vaccine strain recommended for the next influenza season, A/New Caledonia/20/99. For the influenza season 2000/'01, it is recommended by the World Health Organization that the vaccines contain the following (or similar) virus strains: A/Moscow/10/99 (H3N2), A/New Caledonia/20/99 (H1N1) and B/Beijing/184/93. PMID- 11048561 TI - [Acute liver failure ascribed to nefazodone: importance of 'postmarketing surveillance' for recently introduced drugs]. AB - A 50-year-old man developed acute liver failure 7 months after nefazodone treatment was initiated. There was no evidence of any aetiology apart from the exposure to the antidepressant drug nefazodone, while the results of repeated histological examinations of the liver were compatible with serious progressive drug-induced hepatitis. This diagnosis was initially disregarded because in the literature no patients with nefazodone-induced hepatitis were reported. However, the drug had only recently been introduced, which meant that relatively infrequent adverse drug reactions might not have been published. Further analyses of databases of the marketing pharmaceutical industry and of the World Health Organization revealed more cases of liver toxicity ascribed to nefazodone, which showed that liver failure may indeed be associated with nefazodone use. Consequently, prescribing doctors have been warned by the pharmaceutical company about this possible adverse drug reaction. PMID- 11048565 TI - A case of fibrous histiocytoma of the liver. AB - Malignant fibrous histiocytoma (MFH) is a rare disease. We describe a 68-year-old man admitted to the hospital because of malaise. On admission, hematologic and serum chemical examinations showed no abnormalities. A tumor measuring 6.0 x 6.0 x 5.5 cm was found in segment S6 of the right lobe of the liver. A computed tomographic scan of the abdomen revealed a mass surrounded by a capsule-like region with a nonuniform shadow at its margin. The mass contained a nonuniform low density area. A magnetic resonance imaging scan showed low intensity on T1 weighted images and high intensity on T2-weighted images. An angiogram of the abdomen revealed a tumor with a darkly stained margin during the venous phase. Partial resection of the liver, including S6 and part of S7, was performed. On histopathological examination, this case was characterized by a storiform pattern. The inside of the tumor showed a storiform-pleomorphic pattern with inflammatory cell infiltration and partial mucinous degeneration. On immunohistochemical studies, the tumor cells stained positively for CD6. The diagnosis was MFH. PMID- 11048563 TI - [Added value of positron emission tomography with fluoro-18-deoxyglucose as the tracer (FDG-PET) in clinical problem cases in oncology]. PMID- 11048568 TI - [Enzymatic modification of the functional, nutritional and sensorial properties of soybeans for special feeding]. AB - Production of new protein-based products for special nutrition such as hypoallergenic infant formulas, fortified beverages and nutraceutics, require ideal ingredients. Protein ingredients were developed by enzymatic hydrolysis and methionine synthesis of soy protein. Hydrolysis was done at 4% (w/v) using porcine pancreatic enzymes (4% w/w), 50 degrees C, 6 h and pH 8. After drying powder was resuspended (20% w/v) and incubated with 7.6% (w/w) methionine methyl ester, 1% (w/w) chymotrypsin and 3 M glycerol, 37 degrees C, 3 h and pH 7. Hydrolysates were fractionated by ultrafiltration (UF) before and after enrichment (E): FI > 10, 10 > FII > 3 and 3 > FIII > 1 kDa. Functional properties, amino acid content, anti-physiological factor activities and antigenicity were assayed for all the UF fractions and the soybean meal. Protein quality bioassay and sensorial test of an non-enriched fraction and an enriched fraction were performed. Functional properties were positively modified by hydrolysis and synthesis by using a minimum time and methionine added for the last reaction. After UF all the fractions under 10 kDa showed 100% solubility (pH 4 and 7), good clarity, acceptable foam capacity and negligible antigenicity and antiphysiological activities. Additionally, methionine enrichment enhanced their nutritional value, upgrading sulfur amino acid requirements for infants and adults. Because functionality and nutritional value FIII-E could be used for hypoallergenic infant formulas, FII-E for fortified soluble formulas and nutraceutics and FI-E for a semi-solid baby food. PMID- 11048571 TI - [Effect of the lactose induced diarrhea on macronutrients availability and immune function in well-nourished and undernourished rats]. AB - In this study we compared the availability of nutrients in a balanced diet offered to young well-nourished and undernourished Sprague Dawley rats, with and without diarrhea. Malnutrition was induced by restricting food intake (50%) in one half of the rats for 2 weeks and diarrhea was induced by including 45% lactose in the diet after malnutrition had been established. During the experiment which lasted 8 d the animals were kept on the same feeding protocol but one half of the nourished and one half of the undernourished received lactose to induce diarrhea. The results showed that the inclusion of lactose at 45% in the diet caused a severe diarrhea both in the nourished and undernourished rats. This diarrhea however, resulted in a reduction in food intake and growth only in the well-nourished rats. In the rats with diarrhea the apparent digestibility of the diet and of its macronutrients decreased compared with the animals without diarrhea but this reduction was less apparent in the undernourished rats. Similar results were obtained in relation to the retention of nitrogen and energy. In this case, diarrhea was associated with retentions which were lower than those seen in the rats without diarrhea but the undernourished rats with diarrhea retained more energy than the well-nourished rats with diarrhea. Malnutrition resulted in lower packed cell volume, leukocyte count and thymus weight but diarrhea in the malnourished rats did not cause a further reduction in these variables as it did in the well-nourished animals. In general, these results indicate that in well-nourished rats, diarrhea had a negative effect whereas in the undernourished group it did not. It appears that the undernourished rats compensated their nutrient utilization so that diarrhea did not worsen their undernourished condition. PMID- 11048569 TI - [Evaluation of the effects of a school breakfast program on attention and memory]. AB - The purpose of the study was to evaluate the impact of a school breakfast program in Sonora, Mexico. The study was conducted in four counties classified as extremely poor. Three hundred children from twelve schools pertaining to three counties under research received a school breakfast; one hundred and fifty children from four schools pertaining to a fourth county did not received school breakfast. Children were four to six years old. Attention, memory and cognition tests were applied before and after the program. Results showed an improvement for those groups receiving the breakfast program, specially on response speed and behavior executions. Group without breakfast showed better results on the pre test, nevertheless, on the post-test, performances in both groups appear the same. At the beginning of school courses any county was different, but speed in selection and running show significatives differences for the schools. The program shows similar results on response speed, no matter social condition, nevertheless, program differentially affects the number of correct behavior executions on stimulus selection and reproduction. The breakfast program benefit children, but its effects are differentially distributed improving behavioral, repertories depending of children's group vulnerability. PMID- 11048566 TI - [Nutritional importance of phenolic compounds in the diet]. AB - Phenols are one of the major groups of nonessential dietary components appearing in vegetable foods. They are a wide chemical compounds group that are considered as secondary plant metabolites, with different activity and chemical structure, including more than 8,000 different compounds. Phenols, has traditionally been considered as antinutritive compounds due to the adverse effect of one of their main components, tannins, on protein digestibility. However, actually there is an increased interest in these compounds because they have been associated with the inhibition of atherosclerosis and cancer. The bioactivity of phenolics may be related to their antioxidant behaviour, which is attributed to their ability to chelate metals, inhibit lipoxygenase and scavenge free radicals. This review make a global view on the main phenolic compound groups, their organoleptic effects in vegetable foods, their physiological effects in humans, their metabolism, bioavailability as well as their content in the diet. PMID- 11048570 TI - [Use of knee height to correct the body height of elderly Hispanics]. AB - Loss of stature in certain elderly subjects can be attributed to diseases such as osteoporosis, as well as to age and generational effects. In addition, many elders cannot stand straight for accurate measurement. For these cases, total height can be estimated with regression equations based on knee height. The aims of this study were, firstly, to evaluate the applicability of regression equations based on knee height for estimation of stature and, secondly, to document the differences between measured and estimated height in a group of elderly Hispanics with postural problems (n = 166) in comparison with a group of elderly Hispanic without postural problems (n = 270). Using both, estimated and measured height, we also calculated the body mass index (BMI) of both groups of elders. Statistical analyses were done with paired t-tests, within sex and study group. Within the group with postural problems, estimated height was higher than the measured height for both men (p < or = 0.001) and women (p < or = 0.001). There were no significant differences between measured and estimated height in the group without postural problems. Furthermore, in the group with postural problems, BMI values calculated with estimated height were lower than those estimated with the measured height, and these differences were also significant for both men (p < or = 0.001) and women (p < or = 0.001). With the aging of the Latin American population, there is a need for more nutrition and health research among elders. In order to do this we need to develop and use methods and criteria appropriate for each population. PMID- 11048573 TI - [Optimization of formulations for dietetic pastry products]. AB - Optimized formulations of dietetic pastry products such as cake and sponge cake premixes were formulated using the surface response methodology. % Emulsifier agent and baking time were the selected independent variables for cake, as well as % emulsifier agent % chlorinated flour the variables selected for sponge cake. Three different level of each variable summing up thirteen experimental formulae of each product were assessed to optimize the variables that could have some influence in the sensory characteristics of these dietetic products. The total sensory quality was determined for both dietetic products using the composite scoring test and a panel of 18 trained judges. Looking at the contour graphic and considering economic aspects the best combination of variables for cake formulation was 2% emulsifier agent and 48 minutes for baking time, With respect to sponge cake, the best combination was 6% emulsifier agent and 48% chlorinated flour. Shelf life studies showed that both dietetic formulations remained stable during storage conditions of 75 days at 30 degrees C. During this period, significant differences in sensory characteristics were not found (p < 0.05). Data of peroxide values were kept under the critical value reported for detection of organoleptic rancidity. Reported values of hedonic test showed that these dietetics pastry products had good acceptability, and open up marketing opportunities for new products with potential health benefits to consumers. PMID- 11048572 TI - Dietary proteins on reproductive performance in three consecutive generations of rats. AB - The cumulative effects of the long-term consumption of the Regional Basic Diet (RBD) of Northeast Brazil on gestational and lactational performances were assessed in three consecutive generations of Sprague Dawley rats (n = 1,334). The animals were distributed into three groups: RBD (8% predominantly vegetable-based proteins), Control (8% casein) and Standard (22% casein). Primiparous fertilized rats aged 120 days old and their offsprings made up generation 1. Consecutive generations were obtained by mating products from previous generations. Statistical differences between groups and generations were analysed by the method of Kruskal-Wallis. In comparison with their respective controls, data for RBD-fed groups were as follows: decreased conception rates, gradual declines in gestational body weight gains and reduction in the mean number of youngs per litter; the weight loss of lactating rats aggravated in consecutive generations; suckling rats had lower values for birthweight and weight gain, higher values for death rates (including soon after weaning) and severe immaturity at weaning (21 days). This dietary experimental model proved to be valid in terms of providing information for further studies about the relationship between quality and quantity of dietary proteins in order to allow decisions on supplementary feeding programmes for people now most in need. PMID- 11048567 TI - [Training of sensorial panels consisting of children]. AB - In the development of food products for children, it is advisable to establish the characteristics of the product with groups of children that represent the target population. To ensure the success of the products, the quality and hedonic satisfaction expectatives must be considered. In order to accomplish this premises, a group of children under the Program of Complementary Feeding of the Health Ministry--was selected and trained. The project was developed with a group of 33 children ages 9 to 12 years--from the Republica of Colombia School of Santiago, whose parents agreed and supported the participation of their children in this project. The first step was teaching the technics and methodology of Sensory Evaluation, and increasing their sensitivity. After the 8 programmed sessions, those children who met the minimal requirements for a training group were chosen. The second step was performed during 12 sessions, working with 14 children. The training was aimed at the development of the vocabulary to describe quality and defects, ranking tests, discriminative tests and the use of different scales. Tests to verify reliability, veracity and reproducibility of judgements (p < 0.05) were carried out. The trained group was able to assess different meals of the Program. The obtained results allowed to propose the improvement of some quality criteria of the Program meals. PMID- 11048574 TI - [Characteristics of Canavalia proteins]. AB - The purpose of this work was the isolation and characterization of grain protein from five Venezuelan Genotypes (U-02, Yaracuy, Valle De La Pascua, Originally Tovar) of Jack Bean (Canavalia ensiformis). The average protein content from these genotypes was 31.37%, it ranged between 28.44% and 33.05%. The protein isolation was performed by solubility extraction procedures, showed: 84.57% of albumins, globulins and non proteic nitrogen and 15.43% of alcohol insoluble reduced glutelin (AIG). The content of anti-nutritional factors (canavanine and hemagglutination title) found in protein fractions were respectively: Albumins 1.96%, +4; globulins 0.17%, +5 and AIG 0.22%, +1. It was observed that protein fractions of genotype Tovar had the lowest canavanine values (0.79%, 0.02% and 0.00% respectively). The globulins gave the highest in vitro protein digestibility (65.20%) followed by Albumins (58.90%) and AIG (37.28%). PMID- 11048575 TI - [Electrophoretic study of albumins and globulins of 4 genotypes of Canavalia ensiformis]. AB - Polyacrylamide gel electrophoresis (PAGE) and PAGE-SDS were used to study seed albumins and globulins of Canavalia ensiformis. In PAGE the concentration of acrylamide used in the upper gel was 4.0% with a pH of 6.7 and in the lower gel 7.5% and 10% of acrylamide were used with a pH of 8.9. In PAGE-SDS the concentration of acrylamide was 4.4% with a pH of 6.8 in the upper gel and 7.5% and 12.6% with a pH of 8.8 in the lower gel. The material used were the Venezuelan genotypes Tovar, Yaracuy, Original and U-02. The albumins and globulins were extracted with a 0.5 M NaCl solution and then separated by dialysis against water and lyophilized. These protein fractions represented 84.85% of the total amount of protein in seeds. The albumins were separated in PAGE with 7.5% acrylamide into five fractions and globulins into six, with similar electrophoretic patterns between genotypes. In a similar manner, the patterns obtained in PAGE with 10% acrylamide were the same for all genotypes, showing five bands for albumins and three bands for globulins. With PAGE-SDS containing 7.5% of acrylamide, albumins were separated into as many as eight components, and globulins into as many as seven bands with mobilities between 0.2981 and 0.9932, with different patterns for each genotype. Also the patterns PAGE-SDS at 12.6% of acrylamide were different for the genotypes, separating proteins into a greater number of bands. The albumins showed as many as twenty one bands with mobilities between 0.2603 and 0.7398, and globulins as many as sixteen bands with mobilities between 0.2454 and 0.7390. The PAGE patterns of the genotypes analyzed did not show differences between them. However, with PAGE-SDS different electrophoretic patterns were obtained which varied in the number and intensity of the bands, making it possible to distinguish the genotypes studied. The molecular weight of the albumins varied between 76,000 and 12,000 daltons and of the globulins between 80,000 and 12,000 daltons. PMID- 11048576 TI - [Elaboration of an yogurt made of a milk and chickpea (Cicer arietinum) mixture]. AB - The objective of this work was to establish the experimental conditions for the production of yogurt extended with chickpea (Cicer arietinum), inoculated with St. thermophilus and L. bulgaricus and compare its chemical, microbiological and sensorial characteristics versus a yogurt made of skimmed milk. Results indicated that 70:30 and 80:20 (skimmed milk and chickpea extract) mixtures obtained by chemical score fulfilled with the proposed objectives. Yogurt made with 70:30 mixture added with modified starch (ULTRA SPERCE M and COL-FLO), did not remove syneresis present in these products and did not improve its sensory characteristics neither. Nevertheless, yogurt made with 80:20 mixture and modified starch (ULTRA SPERCE M) removed syneresis and present flavor and texture characteristics alike yogurt made of milk, this "extended" yogurt was accepted by the 80% of the judge and fulfill with yogurt specification established in the mexican regulations for this products. PMID- 11048577 TI - [Nitrates and nitrites in homemade and industrial cheeses commercialized in the Southern region of Minas Gerais, Brazil]. AB - The evaluation of nitrate and nitrite in homemade and industrialized cheeses commercialized in the southern region of Minas Gerais State, was the aim of this work. In Brazil, the use of these additives is permitted at maximum levels of 50 mg/kg. The basis of the previously validated method is the quantitative reduction of nitrate to nitrite through cadmium column and spectrophotometric determination after nitrite diazotation with sulphanilic acid/alpha-naphtol reagent. From all samples analyzed, 38 (88.37%) showed neither a nitrate nor nitrite detectable content; 5 samples (11.63%) presented nitrate, 4 of them being above the tolerated level and only one (2.33%) showed detectable nitrite, however below the permissible limit. The majority of samples did not show nitrate or nitrite at detectable levels. However, though probably safe from a toxicological point of view, the results can implicate an increased risk of bacteriological contamination. PMID- 11048578 TI - [Chemical composition of food products derived from wheat and corn produced in Costa Rica]. AB - Twenty one wheat and corn based food products elaborated in Costa Rica were analyzed by chemically with the purpose of having data on local foods. The analytical methods to determine proximate composition were AOAC's. Energy was estimated by calorimetric bomb and dietary fiber (DF) by the gravimetric enzymatic method. Also food portion size was estimated and related with DF content for food classification. The values of the nutrients per food were established and compared with others reported in foreign tables commonly used in the country. Fat and energy content in cookies are higher than in salad breads and crackers. Wheat and corn based food products are classified either as low or very low DF sources (< 2.9 g FD/portion). Corn "tortilla" DF content duplicates bread's and the fiber is basically insoluble. Marked differences were founded in the nutritive composition of specific foods when compared with values reported in foreign food tables. In other foods, as corn based products, similarities in the chemical composition were common. The chemical composition of the studied local foods shows the potential of the diet to be atherogenic, an important aspect to be considered with relation to the main causes of mortality in Costa Rica population. The more compatible food composition table with our data is the Central American, followed by the Latin American one. The necessity of having data on the chemical composition of local foods has been demonstrated. PMID- 11048579 TI - [Dietary fiber in vegetables cultivated in Chile]. AB - The objective of this study was to determine the soluble, insoluble, and total dietary fiber contents in vegetables produced in Chile. The analyses were conducted in the vegetables in the same conditions as they are consumed. Thirty three vegetables obtained from local markets and the Instituto de Investigaciones Agropecuarias (INIA) were studied. Water and fiber contents were determined in 16 raw and 17 cooked (boiled) samples. Moisture range between 63.2% and 96.2%. Average (+/- sd) total, soluble and insoluble contents, expressed as g% on wet basis were: 3.00 +/- 1.59, range 0.96 7.3; 0.93 +/- 0.50 range; 0.30 2.60, and 2.06 +/- 1.26, range 0.51-5.90 respectively. On dry weight basis total fiber concentration was 30.1 +/- 12.5, with a proportion of 68.5% and 31.5% of the insoluble and soluble form, respectively. On dry weight basis total fiber concentration was 30.1 +/- 12.5, with a proportion of 68.5% and 31.5% of the insoluble and soluble form, respectively. Dietary fiber supply ranged between 1.0 to 10.7 g in the large serving sizes. The ratios insoluble fiber/crude fiber and total dietary fiber/crude fiber did not present constant results. Values ranged between 1.1 and 4.5 (mean 2.2 +/- 1.0) in the former, and from 1.4 to 6.5 (mean 3.2 +/- 1.5) in the latter. We conclude that both soluble and insoluble fiber vary widely among vegetables produced in Chile. This study provides information on the fiber composition of vegetables. Such information may help to choose them according to these variables in order to be used in the prevention or treatment of selected pathologies. PMID- 11048580 TI - [Mineral composition of edible mushrooms cultivated in Brazil--Pleurotus spp and other dehydrated species]. AB - Macrofungi can accumulate some minerals, including toxic metals if present in the substrate. A periodic monitoring of these elements in mushrooms is recommended when the conditions of cultivation are altered. The aim of this work was to evaluate the mineral content of Pleurotus spp (hiratake and shimeji) and of imported (chilean and italian) dehydrated mushrooms. Fresh fruiting bodies of Pleurotus spp were obtained from cultivators and dehydrated mushrooms were bought in a market. The samples were dried, milled and digested by C1H-NO3H. The content of P, K, S, Ca, Mg, Cu, Fe, Mn, Zn, Na and B were analyzed by ICP-AES and Al, Cd, Cr, Pb, Co, Ni by ICP-OES. The results classify these mushrooms as a source of potassium and copper: Pleurotus spp are also a source of phosphorus (P < 0.05); the chilean mushrooms present high content of iron (P < 0.05). All the evaluated mushrooms were identified as a food without sodium (< 5 mg Na/100 g). So these mushrooms being a source of potassium without Na, answer the needs of hypertension and/or heart diseases patients as a food and/or like a condiment for flavor enhancement. Subsequent studies should include major sampling and the evaluation of the toxic metals, Pb and Cr, employing more accurate methods of analysis, as well as the evaluation of Hg (not analysed in this study), mainly in wild mushrooms, commercialized dehydrated. PMID- 11048581 TI - [Cariogenicity and cariostatic properties of different types of milk-review]. AB - The purpose of this study is to introduce some information about local and systemic effects of different kinds of milk in oral health, through the explicitness of its cariogenicity and cariostatic properties. Different kinds of milk and milk products are consumed most commonly by the worldwide population, raising the interest of researchers in its influence in the oral health for some decades. Several studies have been conducted to associate the consumption of bovine-, human- and infant formula-milk with caries development and other dental defect, but controversial results have not been able to define the cariogenic and cariostatic potential of different kinds of milk. Bovine milk has some cariostatic components as casein, lipids and antibacterial enzyme, though it has 4% lactose, supposedly cariogenic sugar. Human milk has been related to a sort of caries which is like bottle caries, even though some studies have demonstrated its non cariogenicity. The infant formula milk, which is elaborated for specific period in the childhood, has received low control on its potential for developing caries. It could be of great value to elucidate the controversy surrounding the cariogenicity and cariostatic properties of different kinds of milk, concerning caries prevention during infant and adult life. PMID- 11048582 TI - [Risk factors for growth retardation in children of low economic and social level in Sao Paulo, Brazil]. AB - Aiming to analyze the mild to moderate growth retardation associate factors in children at school age, a case control study was conducted with 153 pairs of seven and eight years old children from public schools of the outskirts of the city. The cases were defined as children with a height for age(H/A) between -1 and -2 Z score, according to NCHS/WHO standards, and matched with a H/A +/- 0.5 Z score children of same age, gender, school, classroom and class time, as controls. The height was measured at school according to WHO rules by trained professionals, and household visits were carried out to obtain environmental and socio-economic data. Initially the data was analyzed by the univariate conditional method and then, the statistically significant variables were included in a model of multivariate logistic regression analysis. As a result, the risk factors remaining at the end of the multivariate analyses, by Odds Ratio and it's 95% Confidence Interval were respectively:--mother's height: for each decrease of one standard deviation the OR increased 1.84(CI: 1.35 to 2.49); inadequate feeding pattern: OR = 2.12; CI: 1.17 to 3.83, very low socio-economic level: OR = 9.2; CI: 3.35 to 25.13, low birth weight: OR = 2.59; CI: 1.44 to 4.63 and smoking during pregnancy: OR = 1.75; CI: 0.98 to 3.12. These results highlight the environment as a determinant factor for growth performance during the first years of the child's life. Despite this, the significant OR for Mother's height allows the assumption that besides the environment, the parent's height has to be considered as one of the determinants of height deficit, even for the low socioeconomic level. PMID- 11048583 TI - [Nutritional evaluation of Lepidium meyenii (MACA) in albino mice and their descendants]. AB - The Maca (Lepidium meyenii) is a Peruvian hypocotyl that grows exclusively between the 3700 and 4500 masl at the Peruvian Andes. Traditionally it is attributed nutritional, energizing, fertilizing properties among others. With the purpose of evaluate scientifically the nutritional property of Maca, we carried out a controlled study in two generations of albino Swiss mice (parents and breeding). The parents were aleatorily assigned to one of three nutritional schedules. The food of each group was prepared based on powder from a commercial balanced food (CBF) of which 30% was replaced by raw or cooked Maca according to the corresponding group or pure CBF in the control group. The groups were this way: 1) Raw Maca Group; 2) Cooked Maca Group; and, 3) Control Group. The results showed that the curves of growth were similar and adequate for the three groups. However, the cooked Maca group showed the best curve. These data were better observable in the second generation of animals, with significant statistical difference (p < 0.05). The CBF group had a better growth than raw Maca group. No signs of malnutrition nor overweight were observed in none of the groups. The serum values of total proteins and albumin were statistically superior for the mice group eating cooked Maca than that of the raw Maca and CBF groups. This study demonstrates, in a scientifical evaluation, one of the traditionally attributed properties of Maca, the nutritional capability. PMID- 11048584 TI - [Low cholesterol egg powder. Nutritive and quality characteristics of the product]. AB - The purpose of this study is to obtain a low cholesterol egg powder for the preparation of different foods for persons whose egg consumption is restricted. Egg white and yolk mixtures prepared in different proportions were dehydrated; the following dried mixtures were obtained: A (1:1), B (2:1) and C (3:1) of egg white:yolk respectively. These mixtures were evaluated using the following parameters: proximal analysis, microbiological assay and protein quality evaluation. Physical characteristics of the powder and the sensorial tests of foods prepared with these mixtures were carried out. The fat and the cholesterol content in the mixture C were decreased by 40% and 20% respectively. The microbiological tests showed that the three mixtures were safe for human consumption. The PER of sample A (whole egg) was 3.65 and for the mixture C 3:1 egg white:yolk was 3.05. The PER of the 50:50 protein mixtures eggs white and yolk: with corn lime treated flour (HMN) were higher than that of the casein standard. The sensorial tests of the foods prepared with all the mixtures were acceptable. PMID- 11048585 TI - [Nutritional and microbiological profile of soybean sausages available in Costa Rica]. AB - The nutritional and microbiological quality of 80 soybean sausage samples (50% frankfurter and 50% sausage mortadela) was studied. On average, the protein content was 17.5 g/100 g in sausage mortadela and 20 g/100 g in frankfurter. The mean total fat content was 5.5 g/100 g for both products. However when products of different manufacture industries were compared, a highly significant difference (p = 0.0000) in the fatty acids speciation between both groups and between samples of the same product were found. Bigger differences were found in the content of palmitic acid (C16:0), oleic acid (C18:1) and linoleic acid (C18:2). Cholesterol was not detected in samples analyzed. On average the atherogenicity index was 0.55 for sausage mortadela and 0.59 for frankfurter. A consumption of 25 grams of of soybean protein from these sausages can bring an intake of saturated fatty acids between 20-90% of the daily recommendation. Likewise, they can supply between 12-70% of the recommended daily polyunsaturated fatty acids. These variations are owing to the big difference in fatty acids speciation in each sausage brand. Around 20% of soybean sausages studied showed total coliform levels above 10(4)/g, being more frequent in sausage mortadela. Also 60% of this product and 10% of frankfurters showed psychrotroph levels of 10(6)/g. Clostridium perfringens, in levels above 10(2)/g was evidenciated in 5% of samples, Escherichia coli was not isolated from them. The findings of this study suggest the urgent need for implementing a quality control system for soybean sausages, before national health authorities consider to support nutritional campainings that promote their consumption. PMID- 11048586 TI - [Use of mesquite cotyledon (Prosopis chilensis (Mol) Shuntz) in the manufacturing of cereal bars]. AB - Cereal bars with peanut and walnut has shown to be snack foods of good organoleptic characteristics and high caloric value, due to their content of protein, lipids and carbohydrates. Cotyledons of mezquite seeds have a high protein content which biological quality improves with thermal processing like toasting, microwave or moist heat under pressure. The purposes of this research were to study the use of mezquite cotyledon (Prosopis chilensis (Mol) Stuntz) in cereal bars with two different levels of peanut or walnut; and to determine the effect of two thermal treatment applied on the cotyledon upon the bar characteristics. Twelve different kind of bars were developed through the combination of two levels of peanut or walnut (15% and 18%); the use of mezquite cotyledon (0% and 6%); and the application of two thermal processing to the cotyledon (microwave and toasting). Cereal bars were analysed for chemical, physical and sensory characteristics: moisture, water activity, proximate chemical composition, sensory quality and acceptability. Moisture content of bars with peanut ranged between 10.4% and 10.9%; and for those with walnut, between 10.5% and 12.3%. Protein content was higher in the bars with mezquite cotiledon, being higher those with peanut. Thermal processing did not have any effect on the chemical composition. Bars with mezquite cotyledon treated by microwave showed a higher acceptability. PMID- 11048587 TI - [Storage of cereal bars with mesquite cotyledon (Prosopis chilensis (Mol) Stuntz)]. AB - The use of walnut or peanut in the elaboration of cereal bars represents a possible risk of undesirable changes during their storage due to their high content of unsaturated fatty acids in the oil; oxidizing of the fatty acids is one of the main causes of deterioration. Development of new snack products implies the use of packages that should protect the food against the damage caused by light and reduce the oxygen concentration of in their interior. The objective of this investigation was to evaluate the physical, chemical and sensory changes in the storage of cereal bars with peanut or walnut and mezquite cotyledon subjected to two thermal treatments, packed in cellophane or milky polypropilene. Four types of bars were elaborated with 6% of mezquite cotyledon, treated by microwaves or toasted, and with 18% of peanut or walnut. The bars were stored for 90 days at room temperature; and each 30 days it was measured moisture content, peroxides index, water activity, sensory quality and acceptability. The peroxides values (4.9-13.8 meq/kg of oil) indicates that the shelf life of the bars in all the studied treatments was 90 days. The packaging materials used allows to maintain in good conditions, for 3 months, the cereals bars of moisture (7.4-11.2%), water activity (0.50-0.65) and sensory acceptability. PMID- 11048588 TI - [Obtaining a fermented chickpea extract (Cicer arietinum L.) and its use as a milk extensor]. AB - Chickpea (Cicer arietinum L) is cultivated in the North part of Mexico and it is considered a good source of vegetal protein of low cost (20% average), nevertheless, the 80% used for the exportation and only the 20% less was used for animal feeding. The main objective in this study is to obtain a fermented chickpea extract for using in dairy extensor. Chickpea water absorbtion kinetics were carried out in e temperature conditions:while the conditions were established, chickpea was grounded and fermented in different amounts with its natural flora, L. casei, L. plantarum and a mixture culture of both microorganism in logarithmic phase. The results showed that the presence of microorganism of chickpea natural flora interferes during the fermentation, so before the inoculation it was necessary treat the chickpea extract (CE) terminally in a dilution 1:4 during 20 min at 7.7 kg/cm2 of pressure. The use of a mixture culture of 5% of L. casei and 5% L. plantarum inoculated in MRS broth was used to decrease fermentation time. Its addition in logarithmic phase to the sterile chickpea extract increased the lactic acid production and decreased the pH value in 6 h which was less time that one obtained with each of lactobacillus. The fermented extract obtained finally, presented similar sensory characteristics to the ones of dairy products. Therefore, chickpea is a good alternative as a extensor for this kind of products. PMID- 11048589 TI - [Vegetable food resources with agroindustrial potential from Guatemala. Manufacture of vegetable milk from the seed of morro fruit (Crescentia alata)]. AB - The objective of the present study was to establish the conditions for the preparation of vegetable milk from the morro or jicara seed, and to characterize the products, the milk and the residue for their partial chemical composition and acceptability. From the ripe fruit, the seeds were obtained by maceration in water for 3.5 hrs, obtaining seed yields of 80%. This seed contained 38% fat and 26% protein on a dry weight basis. The harvested seed was then dehydrated to 9 12% moisture by exposure to solar energy. A similar lot was lightly roasted by heating on a hot surface for 10 min at 90-110 degrees C. The seed was used for the extraction of solubles or the vegetable milk. Water extraction with up to 10 minutes of mechanical blending gave low yields of soluble solids (4.66 +/- 0.10 to 4.98 +/- 0.07%) and low contents of fat, protein and ash in the extract with sun dried seed and significantly lower with the lightly roasted seed (3.0 +/- 0.05 to 3.4 +/- 0.03%). Due to the low yields of total soluble solids and to the low nutrient content by aqueous extraction, new extractants were used consisting of buffer solutions at pH 7.8, or 8.5 with and without saline solution at 0.5% concentration. With these solutions greater amounts of solids were extracted with a higher content of nutrients. The milk prepared from the sun dried seed and extracted with buffer at pH 8.5 and saline solution (0.5%) gave an extract with 9.85% of total solids, 3.37% protein, 4.44% fat. The extraction with roasted seed gave significantly lower yields. The residue of the extraction contained 21.47% fat and 14.72% protein. With the use of buffers and saline solution the extracts had better acceptability, with the milk produced from sun dried seed having better organoleptic characteristics in comparison with soy milk (5.84 vrs 3.76), however it was of lower acceptability when tested against cows milk (5.7 vrs 7.7). The isoelectric point of the extracted protein was between 4 4.45. PMID- 11048590 TI - [Microbiological controls and control points in a hake fillets manufacturing process for exportation]. AB - Indicator and foodborne pathogen microorganisms in the "for export" hake fillets manufacturing were investigated in this study. Critical control points were identified and prevention activities and control were proposed during seafood elaboration process. 45 samples of hake from sequential processing operation stages, 15 ice samples and 12 water samples from utensil washing, were collected. The samples were analyzed for their content of aerobic mesophilic bacteria, psychrotrophic bacteria, enterobacteria, total and fecal coliform bacteria, Staphylococcus aureus and the presence of Escherichia coli, Salmonella and Shigella. The analysis of the samples collected from the factory revealed that the amounts of aerobic mesophile bacteria increased during manual filleting and packaging, in comparison with raw material. Psychrotrophic bacteria were the predominant microorganisms, specially in hake samples. In addition, high levels of enterobacteria, which do not occur normally in fish, were detected in raw hake samples. Staphylococcus aureus, Escherichia coli, Salmonella and Shigella, were not isolated from any samples in this study. The goal of this work is to establish microbiological risks in the hake fillets manufacturing process and, therefore to make possible corrective and control actions to assure the quality and safety of seafood. PMID- 11048591 TI - [Assessment of microbiological quality of food served in dining rooms of private enterprises]. AB - The aim of this study was to evaluate foods microbiological quality in food service establishments. A total of 620 food samples were obtained from four establishments and analyzed for aerobic mesophilic (AM), yeasts, moulds, Staphylococcus aureus and Escherichia coli counts and the presence of Salmonella spp. Drinking water, equipment, surfaces, environment and food handlers were also sampled. E. coli was found in raw vegetables (76.2%), cooked vegetables (15.2%), beef and pork (15.9%), poultry (16.7%), fish (11.8%), desserts (27.3%), equipments (57.9%), surfaces and environment (53.6%) and in 21.9% of food handlers. Survey results were compared with the recommended maximum microbial levels. Our results demonstrate the need for the adoption of more effective hygienic measures in this kind of establishment in order to avoid any risk to consumers. PMID- 11048592 TI - [Determination of deoxynivalenol (DON) in wheat, barley and corn and its relationship with the levels of total molds, Fusarium spp., colonization percentage and water activity]. AB - Fifty samples of cereals including 30 of wheat (10 of wheat hard red spring), 10 of wheat soft red winter and 10 of wheat durum amber), 10 of barley and 10 of corn (5 of white corn and 5 of yellow corn) were analyzed to detect and determine by the TLC method, the quantity of deoxynivalenol levels, which is a toxic secondary metabolite produced by Fusarium species. The aw of samples and the internal and external micoflora and Fusarium spp. levels were also investigated. Results showed that the highest grade of infection (12-80%), and the highest count of total molds (3.9 Log UFC/g) were detected in wheat samples, while the highest levels of Fusarium spp. (2.3 Log UFC/g) were detected in white corn. Deoxynivalenol was found in the wheat and barley samples but not in corn. The wheat red winter soft samples showed the highest levels of deoxynivalenol (3.2 ug/g) which is over the limit levels accepted by the FDA. Correlation was not found among count of total molds, Fusarium spp., infestation grade, aw, and deoxynivalenol levels. These results suggest that it is necessary to exert measures to avoid and to control the importation of contaminated cereals with DON levels higher to those allowed. PMID- 11048593 TI - [Proximal analysis, fatty acids profile, essential amino acids and mineral content in 12 species of fishes of commercial importance in Venezuela]. AB - Proximate composition, fatty acid profile, essential aminoacid and minerals were determined in twelve fish species (armadillo, bocachico, cachama, carpeta, corvina, lisa, mero, merluza, pargo, robalo, tilapia and trucha). Proximate analysis: moisture, protein and ash, were performed using AOAC methodology, fat by Bligh and Dyer method, fatty acids by Gas Chromatography, aminoacid using High Performance Liquid Chromatography and minerals by spectrophotometric method. Results showed that moisture varies between 70.49% for Armadillo and 78.64% for Mero, protein between 18.70% for Merluza and 25.53 for Armadillo, ash between 0.94% for Mero and 2.13% for Carpeta and fat between 1.12% for Pargo and 6.15% for Cachama. Unsaturated fatty acids (omega 3) were the most common found for all the spices. Essential amino acids studied were present in all the spices. Tilapia (10.938 g/100 g of fish), Bocachico (9.231 g/100 g of fish) and Mero (8.738 g/100 g of fish) shown greater content of essential amino acids. Phosphorous was the most concentrated mineral with a mean value of 238.13 mg/100 g of fish followed by calcium with 42.11 mg/100 g of fish. It was concluded that all studied species are an excellent source of protein, omega 3 fatty acids and minerals. PMID- 11048594 TI - [Chemical composition and mineral content of legumes and cereals produced in Northwestern Argentina]. AB - Different varieties of bean (Phaseolus vulgaris), corn (Zea mayz), lentil (Lens sp), soya (Glycine max) and wheat (Tricticum sp) were analyzed in order to obtaining data of chemical composition and content of minerals to contribute to the International Food Data System INFOODS in the elaboration of Regional Food Composition Tables and to evaluate the quality of grains. The selected varieties were the ones produced in the northwestern region of Argentina. The beans were divided in five groups, according to their statistical similarity. The pallares variety is the one that presents bigger content of Cu (2.42 mg/100 g), Fe (76.03 mg/100 g) and Zn (6.08 mg/100 g). The samples of corn were divided in three groups, according to their statistical similarity. The leales yellow corn has bigger content of Zn (3.16 mg/100 g) that the other varieties of the region. The 8 rayas white corn is the one that presents bigger content of Fe (11.48 mg/100 g), while the pisingallo yellow corn is that of bigger content of Cu (1.21 mg/100 g). PMID- 11048595 TI - Characterization of cocoa butter extracted from hybrid cultivars of Theobroma cacao L. AB - Cocoa butter is the most important fat used in the confectionery and chocolate industries. The main objective of the present study was to determine the physical and chemical characteristics of cocoa butter extracted from hybrid cultivars belonging to the germplasm bank of the Fondo Nacional de Investigaciones Agropecuarias (National Foundation for Agricultural Research). AOAC methods were used for the assessment of the proximal composition of the beans, physical and chemical characteristics as well as for the fatty acid profile of the fat. It was found that there were statistical differences in the proximate composition of the cocoa beans among the cultivars studied as well as the iodine and saponification indices of the butter. Saturated fatty acids were present in higher proportions than unsaturated fatty acids, with palmitic and stearic acid as the main fractions. Oleic acid content was higher than linoleic acid. The fatty acid profile found is the main factor that influences the hard texture of the cocoa butter from Venezuelan cocoa hybrids cultivars. PMID- 11048596 TI - Medical students going on elective: who has duty of care? PMID- 11048597 TI - Antibiotic-resistant gonorrhoea. PMID- 11048598 TI - Fungi, mycological disease and pathogenic determinants. AB - The rapid evolution of human fungal infections is providing a strong impetus for understanding pathogenesis and host-fungus interactions and hence new diagnostics. PMID- 11048599 TI - Epidemiology. AB - Invasive fungal infections have emerged as important causes of hospital related morbidity and mortality. They are increasingly seen in patients not previously considered at risk, e.g. patients on an intensive care unit. Candida albicans and Aspergillus spp. are the most common pathogens, posing challenges in epidemiology, control and treatment. PMID- 11048600 TI - Clinical and laboratory diagnosis. AB - Clinical diagnosis of invasive fungal infections demands a high clinical acumen, but both under- and over-diagnosis are common. Laboratory mycological tests have traditionally been limited by a range of complications. Development of methodologies to detect fungal-specific immunoglobulins, cell wall antigens and nucleic acid have improved dramatically in recent years. PMID- 11048601 TI - Management. AB - The management of invasive fungal infection utilizes a variable multidisciplinary approach involving antifungals, appropriate surgery and immuno-correction. Conventional amphotericin B has been the mainstay of treatment but newer pathogens and poor outcomes has led to new formulations of this drug as well as to new and novel antifungals. PMID- 11048602 TI - Transanal endoscopic microsurgery. AB - The anatomy of the pelvis makes it difficult to perform local excisions in the rectum when a tumour is some distance from the anal verge. Transanal endoscopic microsurgery, a minimally invasive procedure, has been developed. It provides an alternative to the transsacral or transabdominal approach, with subsequent shorter hospital stay and fewer complications. PMID- 11048603 TI - Renagel: reducing serum phosphorus in haemodialysis patients. AB - Approximately half of all dialysis patients experience persistent hyperphosphataemia, which causes significant problems. Renagel is a calcium-free, aluminium-free phosphate binder which reduces serum phosphorus in haemodialysis patients by reducing absorption of dietary phosphorus. This article reviews the management of hyperphosphataemia and presents some Renagel clinical trial data. PMID- 11048604 TI - Management of venous and cardiovascular thrombosis: enoxaparin. AB - Enoxaparin has strong clinical evidence that supports its license in a broad spectrum of therapeutic indications, including thromboprophylaxis in surgical patients, medical patients bedridden because of acute illness, the once-daily treatment of venous thromboembolism and the treatment of unstable angina and non Q wave myocardial infarction. PMID- 11048605 TI - How accurate is cancer scan reporting? AB - The wide recognition of advances in medical imaging need to be matched by an awareness of the significant error rates in radiological interpretation of cancer scans. Common errors, their aetiology and potential solutions are discussed. Internet awareness of these errors has appeared. PMID- 11048606 TI - Anticoagulant therapy and referral form. Haemostasis and Thrombosis Task Force of the British Committee for Standards in Haematology. AB - Many hospitals use the recommendations contained in the British Committee for Standards in Haematology anticoagulant guideline documents to develop local protocols for anticoagulant management. A combined anticoagulant treatment chart and referral form has been produced to help incorporate the recently updated recommendations for oral anticoagulation into day-to-day practice. PMID- 11048607 TI - Receptor pharmacology of neuroleptics. AB - The term 'neuroleptic' originated from animal experiments--these drugs caused profound sedation and abnormal posturing as if the animal had been 'seized' (Greek 'lepsis', meaning seizure). All early neuroleptic drugs had antipsychotic actions and vice versa. Now there are drugs which are potent antipsychotics without the classic neuroleptic actions known as atypical neuroleptics. PMID- 11048608 TI - Irreversible aromatase inactivation: treatment for breast cancer. AB - Third-generation aromatase inhibitors extend treatment options for postmenopausal breast cancer patients refractory to initial antioestrogen therapy. This article reviews advances and recent developments in hormonal therapy, focusing in particular on exemestane, a new class of aromatase inactivator. PMID- 11048609 TI - Thromboprophylaxis in medical patients. AB - Recent clinical studies have confirmed that acutely ill medical patients are at substantial risk of venous thromboembolism, which can be reduced by thromboprophylaxis with low molecular weight heparin. These studies have resulted in approval of a low molecular weight heparin (enoxaparin) for the prophylaxis of venous thromboembolism in medical patients bedridden as a result of acute illness. PMID- 11048610 TI - Bicalutamide treatment for locally advanced prostate cancer. AB - In men with locally advanced prostate cancer, bicalutamide 150 mg monotherapy provides a similar disease outcome to medical or surgical castration. However, castration is associated with loss of sexual interest and function, decreased energy and an increased risk of osteoporotic fractures. Bicalutamide 150 mg monotherapy has less impact on sexual interest and physical capacity than castration. PMID- 11048611 TI - Both service and training demand attention in obstetrics and gynaecology. AB - New proposals for workforce planning, training and ways of working in the NHS are under consideration. Aspects of service, training and workforce in obstetrics and gynaecology that call out for change include training in the senior house officer grade, especially for GPs and non-EC doctors, and the work of consultants. Proposals for change present a golden opportunity for the specialty to lead in new systems of both service and training, which are closely linked. PMID- 11048612 TI - Recurrent postdural puncture headache. PMID- 11048613 TI - Tuberculosis as a cause of chronic epididymo-orchitis in a male caucasian. PMID- 11048614 TI - The national plan: is that it? PMID- 11048616 TI - Re-emergence of syphilis. PMID- 11048615 TI - Arterial thoracic outlet compression syndrome: a differential diagnosis of painful right supraclavicular swelling? PMID- 11048617 TI - Re-emergence of syphilis. PMID- 11048618 TI - Neurological manifestations of malignant disease. PMID- 11048619 TI - Current strategies for mechanical ventilation in acute lung injury. PMID- 11048620 TI - Design and conduct of clinical trials in patients with osteoarthritis: recommendations from a task force of the Osteoarthritis Research Society. Results from a workshop. PMID- 11048621 TI - Growth factors, insulin-like growth factor-1 and growth hormone, in synovial fluid and serum of patients with rheumatic disorders. AB - OBJECTIVE: Synovial fluid (SF) plays an important role in joint function. We evaluated the growth factors, insulin-like growth factor-1 (IGF-1) and growth hormone (GH) in SF and serum from patients with osteoarthritis (OA), rheumatoid arthritis (RA), gout, pseudogout and diffuse idiopathic skeletal hyperostosis (DISH). DESIGN: Standard radioimmunoassay techniques were used to measure concurrent levels of IGF-1 and GH. SF samples and serum samples were obtained concomitantly from 27 patients with OA, 22 patients with RA, nine men with gout, 14 patients with pseudogout and eight men with DISH. RESULTS: In the case of IGF 1, a comparison of serum and SF levels shows that SF levels of IGF-1 are lower than serum levels in all groups. Men and women gave similar values. In contrast, in the case of GH, all groups, except males with RA, had higher GH values in SF when compared with serum values. Individual patients with other forms of arthritis demonstrated similar relationships. CONCLUSION: The finding that IGF-1 is present in levels about one-half as great in SF as compared with serum suggests that IGF-1 may be produced in lesser amounts or is utilized by the patient in customary joint function. The finding that GH is present in SF at values twice as high, or more, of serum levels in inflammatory arthritides suggests that GH may play a role in the pathophysiology of arthritic disorders. PMID- 11048622 TI - Effect of diacetyl rhein on the development of experimental osteoarthritis. A biochemical investigation. AB - OBJECTIVE: To investigate the effect of diacetyl rhein (DAR) on the synthesis, turnover and composition of cartilage in an experimental model of osteoarthritis in beagle bitches. DESIGN: Osteoarthritis was induced in mature beagle bitches by the transection of the cranial cruciate ligament. Six animals received DAR 20 mg/kg daily for 11 weeks. A matched group received empty capsules daily for the same period. At 11 weeks, articular cartilage was examined for the ratio of the 6:4-sulfated disaccharides of chondroitin and the tissue concentration of hydroxyproline and glycosaminoglycan. In addition, labeling studies were performed to estimate the effect of DAR on proteoglycan synthesis and turnover. RESULTS: DAR had no effect on body weight or food consumption but induced a mild diarrhea and slightly increased the incidence of vomiting. DAR tended to reduce proteoglycan synthesis, however, DAR did reduce proteoglycan turnover in the femoral cartilage. DAR produced changes in the composition of the osteoarthritic cartilage that could only partly be accounted for by changes in hydration and/or swelling. In addition, it was noted that induction of osteoarthritis increased the ratio of chondroitin 6-sulfated to chondroitin 4-sulfated disaccharides; DAR reduced the ratio in tibial plateau cartilage from osteoarthritic joints compared with untreated tissue from osteoarthritic joints. DAR showed moderate reduction on the biosynthesis of proteoglycans. DAR also produced a reduction in proteoglycan turnover from all anatomical areas compared with non-treated controls in both the lateral and medial femoral condyles. CONCLUSIONS: DAR was well tolerated by the experimental animals, but did not produce significant changes in the synthesis or turnover of proteoglycans. The slight reduction in proteoglycan synthesis may prove to be biologically significant after chronic dosing. DAR's effects on the hydroxyproline and glycosaminoglycan content suggest, however, that it must influence the swelling of cartilage and loss of glycosaminoglycan. This indicates that small changes can translate, to significant differences in cartilage composition over an 11-week time period. PMID- 11048623 TI - IGF and IGF-binding protein system in the synovial fluid of osteoarthritic and rheumatoid arthritic patients. AB - Various arthritic disorders result from a disruption of the equilibrium between the synthesis and degradation of tissue matrix macromolecules. Growth factors, particularly insulin-like growth factor-I (IGF-I), are believed to play an important role in maintaining this equilibrium. In this study, we determined the levels of IGF-I, IGF-II, and characterized and measured the amount of IGF-binding proteins (IGFBPs) in the synovial fluid (SF) of osteoarthritis (OA), rheumatoid arthritis (RA) patients and normal individuals. Furthermore, we characterized the IGFBP found in these SFs. The levels of IGF-I, IGF-II and IGFBP-3 were determined by specific radioimmunoassays (RIAs). IGFBP identification and measurement were carried out using the Western ligand blot (WLB) technique, and characterization performed by Western immunoblot. IGFBP-3 proteolysis was analyzed by autoradiography after incubation of SF with radiolabeled IGFBP-3. Results showed a statistically significant increase (P < 0.001) in the IGF-I level in arthritic SF vs normal controls; 75 +/- 11 ng/ml and 82 +/- 11 ng/ml were recorded for RA (N = 8) and OA (N = 10), respectively, whilst normal controls (N = 9) were at 19 +/- 7 ng/ml. No difference in the level of IGF-II was recorded between the three groups studied. Human SF demonstrated the presence of IGFBP-1, -2, -3 and -4, but not that of IGFBP-5 and -6. The level of IGFBP-3 tested either by WLB or RIA was significantly higher (P < 0.001) in RA and OA patients. Moreover, a statistical and positive correlation between the levels of IGF-I and IGFBP-3 was noted. WLB analysis indicated that the amount of IGFBP-1 did not vary among the groups. The levels of IGFBP-2 and -4 were significantly increased (P < 0.02) solely in the RA SF. Further experiments demonstrated that a limited IGFBP-3 proteolysis occurred in human SF. Moreover, the ratio of total IGF over total bioactive IGFBPs was lower in RA (P < 0.05), and to a lesser extent in OA than normal specimens. This study showed the presence of four IGFBPs (1 4) in human SF for which the IGFBP-2, -3 and -4 were enhanced in arthritic fluid. Importantly, although proteolysis occurred in the SF, an increased amount of bioactive IGFBPs were present in arthritic SF, which may affect the bioavailability of IGF-I within the articular tissues. PMID- 11048624 TI - Sequestration of type VI collagen in the pericellular microenvironment of adult chrondrocytes cultured in agarose. AB - The chondron represents the chondrocyte and its pericellular microenvironment and plays an important role in the progression of osteoarthritis. Type VI collagen is preferentially localized in the pericellular microenvironment of adult articular cartilage and increases during osteoarthritis. In this study, we characterized the pericellular sequestration of type VI collagen in long-term chondrocyte agarose cultures, and assessed the action of interleukin-1 on type VI collagen deposition and assembly. Immunohistochemical and biochemical analysis showed that cultured chondrocytes initiate type VI collagen sequestration immediately upon plating and continue pericellular matrix sequestration in a time dependent manner. Confocal microscopy confirmed the cell surface localization and pericellular accumulation of type VI collagen, while image analysis identified a 'cargo-net like' organization of type VI collagen around each chondrocyte. Quantitative analysis revealed a primary phase of rapid cell division and low levels of type VI collagen sequestration, followed by a secondary phase of relative growth stability and high levels of type VI collagen deposition. Interleukin-1 treated cultures showed increased sequestration and retention of type VI collagen in an expanded microenvironment surrounding the chondrocytes. The data suggests a role for type VI collagen in the differentiation of the pericellular microenvironment in vitro. The increased type VI collagen sequestration promoted by interleukin-1 was consistent with previous studies on osteoarthritic cartilage, and implies a functional role for type VI collagen in the chondron remodeling associated with cartilage degradation. PMID- 11048625 TI - Immunohistochemical identification of chodrocalcin in synovial chondromatosis. PMID- 11048626 TI - Reprint of "The Operative Treatment of Communicating Hydrocephalus" by Walter E Dandy, MD. 1938. PMID- 11048627 TI - Acute-onset nontraumatic paraplegia in childhood: fibrocartilaginous embolism or acute myelitis? AB - Fibrocartilaginous embolus causing acute spinal cord infarction is a rare cause of acute-onset paraplegia or quadriplegia. Few cases of survivors have been reported in the neurosurgical literature, with most reports involving postmortem or biopsy findings. There is little information on MRI findings in such patients. We present the youngest patient ever reported, and discuss the important differences between fibrocartilaginous embolus and acute myelitis of childhood. A 6-year-old girl with a history of back pain presented with sudden-onset nontraumatic paraplegia, with a clinical anterior spinal artery syndrome. Initial MRI scan revealed intervertebral disc disease at L1-2 and an incidental thoracic syrinx, but no cause for her acute-onset paraplegia was identified. Cerebrospinal fluid and other investigations were all negative. Sequential MRI scans revealed development of spinal cord expansion from T10 to the conus medullaris, with increased cord signal in the anterior aspect of the spinal cord. The intervertebral disc disease was unchanged. The imaging and clinical findings were caused by fibrocartilaginous embolus, which meant there was no need for spinal cord biopsy. The report describes the clinical and imaging criteria for diagnosis of fibrocartilaginous embolus, highlighting the case for avoiding an unnecessary biopsy. The clinical pattern in the paediatric group is discussed, with features differentiating it from acute myelitis of childhood. PMID- 11048628 TI - Expansion of Chiari I-associated syringomyelia after posterior-fossa decompression. AB - Chiari I malformation (CMI) is an abnormality that involves caudal herniation of the cerebellar tonsils into the foramen magnum. CMI has been shown to be closely associated with the development of syringomyelia (SM). OBJECTS: Several theories have emerged to explain the apparent correlation between the existence of CMI with subsequent development of SM. However, the exact mechanism of the evolution of SM is still subject to controversy. We report here the case of a 12-year-old girl admitted to hospital with headache, vomiting, ataxia, and moderate pyramidal signs. METHODS: Radiological evaluation revealed the presence of CMI, accompanied by a small SM. The patient underwent posterior fossa decompression and improved significantly. She was re-admitted 6 months later with clinical evidence of progressive spinal cord dysfunction. MR revealed gross expansion of the syrinx. CONCLUSIONS: This case raises questions regarding the pathophysiology of CMI and its association with SM. The case indicates the need for neurological and radiological follow-up for patients undergoing posterior fossa decompression due to CMI, even for those without an initial syrinx. This is the first report known to us of expansion of a syrinx following decompression of an associated CMI. PMID- 11048629 TI - The effect of C677T mutation of methylene tetrahydrofolate reductase gene and plasma folate level on hyperhomocysteinemia in patients with meningomyelocele. AB - To evaluate the relationship between genotypes of methylene tetrahydrofolate reductase (MTHFR), and plasma folate and homocysteine (Hcy) levels in meningomyelocele, 21 Korean patients, 47 of their family members, and 43 healthy controls were recruited. The presence of C677T mutation in the MTHFR gene and plasma concentrations of folate/Hcy were investigated. The genotype frequency of C677T mutation was not higher in study groups (patients and family members). The plasma folate concentration showed no difference either between the study and the control groups or among MTHFR-genotypic groups. The plasma Hcy concentration in homozygotes in the study group was higher than that in the control group, and higher than that in heterozygotes when plasma folate levels were low (P=0.006). Although neither MTHFR genotype nor plasma folate/Hcy level plays a definite part on its own, they seem to have an additive effect on the occurrence of meningomyelocele. Our results support folate supplementation for the prevention of hyperhomocysteinemia and meningomyelocele. PMID- 11048630 TI - Histopathological and biochemical changes in the sutural region in craniosynostosis. AB - OBJECT: Histopathological observations and biochemical analysis of sutural bones in nine patients with craniosynostosis were compared with control subjects of the same age range. METHODS: Microscopic examination in craniosynostosis showed formation of an active osseous front, with higher osteoblastic activity than in controls. Biochemical analysis revealed higher calcium, phosphorus, alkaline phosphatase, phospholipids and chondroitin sulphate-A contents in sutural bones of the same patients. CONCLUSIONS: The present study systematically establishes a premature increase in osteogenesis in the sutures of craniosynostosis patients. PMID- 11048631 TI - The use of gated cine phase contrast and MR venography in achondroplasia. AB - OBJECTS: Foramen magnum and jugular foramen stenosis, well-known problems in achondroplasia, may result in brain stem compression and venous outflow obstruction, respectively. We studied a series of children with achondroplasia using gated cine phase contrast (PC) CSF flow studies to evaluate CSF dynamics across the foramen magnum and MR venography (MRV) to depict obstructed venous drainage. METHODS: Ten patients (9 months to 11 years, mean 2.85 years) were referred for possible brain stem compression. MRI included routine sequences, cine PC with velocity encoding (VENC) = 5 cm/s, and MRV. Six patients, including the asymptomatic patient, had brain stem compression without tonsillar herniation; two had tonsillar herniation; and two had neither brain stem compression nor tonsillar herniation. Abnormal tonsillar movement was seen only with tonsillar herniation. MRV showed steno-occlusive disease of the internal jugular vein (IJV) in nine patients, sigmoid sinus in four, and absent or hypoplastic transverse sinus in seven. Veno-occlusive disease was not progressive. No patient had massive hydrocephalus, although larger ventricles were associated with more profuse venous collateral formation and more severe degrees of IJV stenosis. Three patients have undergone CSF diversion. CONCLUSIONS: MR imaging may be useful in defining the pathophysiology of brain stem compression and hydrocephalus in achondroplasia. PMID- 11048632 TI - The expression of inherited hydrocephalus in H-Tx rats. AB - OBJECTIVES: H-Tx rats develop severe hydrocephalus in late gestation. The breeding colony maintained at the University of Florida stems from one pair obtained in 1992. The aims of this study were to characterize the expression of hydrocephalus in the H-Tx rat colony, to perform within-strain and between-strain DNA analysis and to examine hydrocephalus expression in specific breeding experiments. METHODS AND OBSERVATIONS: Matings between normal rats produce hydrocephalic offspring almost without exception, and the overall frequency is stable between generations at 40%. However, frequency varies with parity, being only 27% in the first litters, and it also varies with sex, there being an excess of male hydrocephalics. Mating between shunt-treated hydrocephalic rats did not increase the frequency. DNA typing with microsatellite markers showed that there was some residual heterogeneity in the colony despite inbreeding for 22 generations, although it did not segregate with hydrocephalus. Test mating with two other inbred strains, F344 and LEW produced some affected pups in the LEW cross only. A backcross experiment between H-Tx and F344 produced 12.3% severely affected pups and 5.4% pups with a mild form, indicating the presence of several susceptibility genes. CONCLUSIONS: All animals in our H-Tx colony are homozygous for the hydrocephalus loci, but there is incomplete penetrance. PMID- 11048633 TI - Postraumatic intracranial hematomas in infancy. a 16-year experience. AB - OBJECTS: The objective of this study is to analyze some of the epidemiological aspects in patients with post-traumatic intracranial hematomas (post-traumatic ICH) in infancy. These patients were treated at the Hospital Infantil Municipal de Cordoba, Argentina, between April 1980 and April 1996. METHODS: A retrospective descriptive analysis was conducted on the 113 case histories of children with post-traumatic ICH, all of whom required surgical intervention during this period. Relevant information such as age, sex, mechanism of injury causing craniocerebral trauma (CCT), and data on clinical presentation on admission, diagnosis, morbidity and mortality rates were collected. CONCLUSIONS: The series revealed a greater incidence of post-traumatic ICH in boys (73.5%), whose average age was 6 years 5 months +/- 4 years 10 months (range 1 day to 15 years). Fifty-three percent of the girls suffered post-traumatic ICH before the age of 3, while 54% of the children were 7 years of age or older (P<0.05). Falls were the most frequent mechanism of injury causing CCT (36.3%), followed by vehicle accidents (33.6%) and unknown causes (15.9%), the latter mainly in children under 3 years old (31%). The most frequent symptoms were vomiting (58.6%), loss of consciousness (47.1%) and headaches (24.1%). Of all these children, 93.8% presented signs and symptoms at the time of hospital admission, alterations in the level of consciousness (66%), vomiting (47.2%) and headaches (26.4%) being among the most frequent. The hemorrhagic complications observed in the 113 patients took the form of extradural hematomas (EDH) in 75 (66.4%), of subdural hematomas (SDH) in 35 (31.0%), of hemorrhagic contusions (Hc) in 19 (16.8%), and of intracerebral hematomas (Ich) in 11 (9.7%). In 13 patients the site of the hematoma was the posterior cranial fossa (11.5%), and 22.1% of patients presented more than one type of hemorrhagic complication. Morbidity rates were 9.7% and mortality rates 17.7%. PMID- 11048634 TI - Infantile head injury, with special reference to the development of chronic subdural hematoma. AB - An infantile head injury has unique features in that infants are totally helpless and dependent on their parents, and biomechanical characteristics of the skull and brain are very different from those of other age groups. The authors reviewed a total of 16 infant head injury patients under 12 months of age who were treated in our hospital from 1989 to 1997. Birth head injury was excluded. The most common age group was 3-5 months. Early seizures were noted in 7 cases, and motor weakness in 6. Three patients with acute intracranial hematoma and another 3 with depressed skull fracture were operated on soon after admission. Chronic subdural hematomas (SDHs) developed in 3 infants. Initial CT scans showed a small amount of SDH that needed no emergency operation. Resolution of the acute SDH and development of subdural hygroma appeared on follow-up CT scans within 2 weeks of injury. Two of these infants developed early seizures. Chronic SDH was diagnosed on the 68th and 111th days after the injuries were sustained, respectively. The third patient was the subject of close follow-up with special attention to the evolution of chronic SDH in view of our experience in the previous 2 cases, and was found to have developed chronic SDH on the 90th day after injury. All chronic SDH patients were successively treated by subduro-peritoneal shunting. In conclusion, the evolution of chronic SDH from acute SDH is relatively common following infantile head injury. Infants with head injuries, especially if they are associated with acute SDH and early development of subdural hygroma, should be carefully followed up with special attention to the possible development of chronic SDH PMID- 11048635 TI - A new technique for correction of trigonocephaly in an infant: application of an absorbable endocranial plate. AB - We report on a new method of cranial bone osteofixation using absorbable material, which is applied endocranially to avoid the application of a thick, cosmetically inferior system epicranially in the frontal area. This is especially important in the correction of deformity in trigonocephaly cases. A 9-month-old boy presented with a prominent trigonocephalic deformity. Cranioplasty was performed, in which a supraorbital bar (bandeau) was removed, reinforced with a self-reinforced polylactide (SR-PLA) plate and reshaped to correct the deformity. The plate was applied on the inner side of the bony bar, with its ends fixed epicranially to the temporal bones by means of SR-PLA miniscrews. 3-D CT scans and rapid prototypes were used to evaluate the results. Stable and secure fixation was obtained, and the cosmetic result appeared excellent. No complications occurred during the postoperative period of 1 year. Hence, stabilisation of cranial bone can be achieved endocranially using an absorbable plate and screws, with an encouraging cosmetic result. The method is thought to be reliable. PMID- 11048636 TI - Scalp cavernous angioma presenting as sinus pericranii: diagnostic value of cerebral angiography and magnetic resonance imaging. AB - OBJECTS: Sinus pericranii is only a symptom complex, and it can have a variety of etiologies. Therefore, it is important to differentiate these etiologies preoperatively by means of radiological examinations. A 5-year-old boy was admitted with a soft and fluctuant tumor in the right parietal region near the midline. The tumor appeared when the child was in a recumbent position, distending noticeably with the Valsalva maneuver and disappearing completely when the patient was in the sitting position. METHODS: Magnetic resonance imaging showed the lesion with honeycomb-like heterogeneous iso- and low-intensity signals on the T1-weighted image and with heterogeneous high- and isointensity signal on the T2-weighted image. Dynamic study with an injection of gadolinium diethylene-triaminopentaacetic acid demonstrated and nodular peripheral enhancement at early phase and subsequent progressive enhancement towards the center of tumor. The internal carotid angiogram was normal. The external carotid angiogram, however, showed a tumor stain fed by the superficial temporal arteries. The stain was retained until the late phase and drained into the scalp veins and into the superior sagittal sinus. Following direct injection of contrast medium into the tumor there was prolonged retention of the medium in the tumor and leakage into scalp veins and the superior sagittal sinus. The mass under the periosteum was totally removed and proved to be a cavernous angioma. CONCLUSIONS: Scalp cavernous angioma is one of the etiologies of sinus pericranii and may be diagnosed preoperatively by cerebral angiography or magnetic resonance imaging. Serial dynamic magnetic resonance imaging will be particularly helpful for this diagnosis. PMID- 11048637 TI - Thoraco-lumbar duplication of the spine. Case report and embryology review. AB - OBJECT: We present a case of an asymptomatic and neurologically normal 6-year-old girl who was noted to have a gross spinal abnormality. METHODS: Neuroimaging demonstrates gross thoraco-lumbar spine duplication, which is at the severe end of the split cord malformation (SCM) group of congenital abnormalities. Despite the position of the neural structures, the clinical condition has so far not suggested that any surgical intervention is indicated, though the girl's later growth spurts may still unmask features of a tethered spinal cord. CONCLUSIONS: We present a brief review of the literature dealing with theories of embryogenesis relating to SCM, the common clinical and radiological features, and finally the surgical options available. PMID- 11048638 TI - Spinal clear cell meningioma presented with progressive paraparesis in infancy. AB - Clear cell meningioma, about 20 cases of which have been reported in the literature, is a morphological variant of meningioma. The authors report a case of spinal clear cell meningioma that occurred in a child. A 14-month-old girl showed gradually progressive paraparesis 1 month after she started to walk. Magnetic resonance image showed an intradural extramedullary mass compressing the conus medullaris and cauda equina. Complete excision of the tumor was done, and the patient gradually recovered from motor weakness and neurogenic bladder. Histological examinations along with immunohistochemical and ultrastructural investigations allowed a diagnosis of clear cell meningioma. During the follow-up period, a recurrent mass lesion was detected on the 8-month follow-up MR image in the same region. Because clear cell meningioma might be biologically aggressive, postoperative adjuvant therapy and close follow-up investigation should be considered. PMID- 11048639 TI - The mechanism of phospholipase C-gamma1 regulation. AB - Phospholipase C (PLC)1 hydrolyzes phosphatidylinositol 4,5-bisphosphate to generate the second messengers, inositol 1,4,5-trisphosphate (IP3) and diacylglycerol (DAG). IP3 induces a transient increase in intracellular free Ca2+, while DAG directly activates protein kinase C. Upon stimulation of cells with growth factors, PLC-gamma1 is activated upon their association with and phosphorylation by receptor and non-receptor tyrosine kinases. In this review, we will focus on the activation mechanism and regulatory function of PLC-gamma1. PMID- 11048640 TI - Differential changes in the expression of cyclic nucleotide phosphodiesterase isoforms in rat brains by chronic treatment with electroconvulsive shock. AB - Electroconvulsive shock (ECS) has been suggested to affect cAMP signaling pathways to exert therapeutic effects. ECS was recently reported to increase the expression of PDE4 isoforms in rat brain, however, these studies were limited to PDE4 family in the cerebral cortex and hippocampus. Thus, for comprehensive understanding of how ECS regulates PDE activity, the present study was performed to determine whether chronic ECS treatment induces differential changes in the expression of all the PDE isoforms in rat brains. We analyzed the mRNA expression of PDE isoforms in the rat hippocampus and striatum using reverse transcription polymerase chain reaction. We found chronic ECS treatment induced differential changes in the expression of PDE isoform 1, 2, 3, 4, 5 and 7 at the rat hippocampus and striatum. In the hippocampus, the expression of PDE1A/B (694%), PDE4A (158%), PDE4B (323 %), and PDE4D (181%) isoforms was increased from the controls, but the expression of PDE2 (62.8%) and PDE7 (37.8%) decreased by chronic ECS treatment. In the striatum, the expression of PDE1A/B (179%), PDE4A (223%), PDE4B (171%), and PDE4D (327%) was increased by chronic ECS treatment with the concomitant decrease in the expression of PDE2 (78.4%) and PDE3A (67.1%). In conclusion, chronic ECS treatment induces differential changes in the expression of most PDE isoforms including PDE1, PDE2, PDE3, PDE4, PDE5, and PDE7 in the rat hippocampus and striatum in an isoform- and brain region-specific manner. Such differential change is suggested to play an important role in regulation of the activity of PDE and cAMP system by ECS. PMID- 11048641 TI - Molecular cloning and sequencing of rat Cdc42 GTPase cDNA. AB - Cdc42 is a member of the Rho family of small GTP-ase and plays an important role in intracellular signaling pathways regulating cell morphology, motility and stimulation of DNA synthesis. We have isolated cDNA encoding Cdc42 from a rat brain cDNA library using PCR-cloning strategy. The sequence of isolated gene revealed an open reading frame of 576 nucleotides with the potential to encode a protein of 191 amino acids with a predicted molecular weight of 21 kD. The resulting sequence was incorporated into the GenBank with accession number, AF205635. Sequence analysis revealed that overall cDNA sequence identity is 96% with human G25K and 52% with rat Chp, a homologue of the GTPase human Cdc42Hs, and having one nucleotide difference from the mouse Cdc42. However, putative protein sequence was identical to the mouse and human brain Cdc42Hs. On expression of the cDNA in COS-7 cells, a protein molecular weight of 21 kD was detected in immunoblotting using anti-human Cdc42 antibodies. Therefore, these results suggest that the cDNA we are reporting is most likely the rat homologue of the GTPase human Cdc42. PMID- 11048642 TI - Molecular assembly of mitogen-activated protein kinase module in ras-transformed NIH3T3 cell line. AB - The ras, is a G-like protein that controls the mitogen-activated protein kinase (MAPK) pathway involved in control and differentiation of cell growth. MAPK is a key component of its signaling pathway and the aberrant activation may play an important role in the transformation process. To better understand roles of ras in the activation of MAPKs, we have established ras transformed NIH3T3 fibroblast cell line, and analyzed the MAPK module. The ras transformed cells formed numerous spikes at the edges of cells and showed loss of contact inhibition. The levels of ERK1/2 MAPKs as revealed by Western blot analysis were not significantly different between ras transformed and non-transformed cells. However, phosphorylation of ERK MAPKs and the level of MEK were significantly increased although the heavily expressed level of Raf-1, an upstream component of MAPK pathway was unchanged in ras transformed NIH3T3 cells. The sedimentation profile of the MAPK module kinases in a glycerol gradient showed the presence of a rather homogeneous species of multimeric forms of ERK1/2 and MEK as indicated by the narrow distribution peak areas. The broad sedimentation profile of the Raf 1 in a glycerol gradient may suggest possible heterologous protein complexes but the identification of interacting molecules still remains to be identified in order to understand the organization of the MAPK signal transduction pathway. PMID- 11048643 TI - Effects of allyl sulfur compounds and garlic extract on the expression of Bcl-2, Bax, and p53 in non small cell lung cancer cell lines. AB - Allyl sulfur compounds play a major role in the chemoprevention against carcinogenesis. The present study compared the antiproliferative effects of diallyl sulfide (DAS), diallyl disulfide (DADS) and garlic extract on p53-wild type H460 and p53-null type H1299 non small cell lung cancer cells (NSCLC). The DAS and DADS treatment of both H460 and H1299 cells resulted in the highest numbers of cells in apoptotic state as measured by acridine orange staining, however, garlic extract treatment did not induce any significant apoptotic cells by MTT assay. DADS was found to be more effective in inducing apoptosis on NSCLC. The level of p53 protein in H460 cell was increased following DADS treatment. DAS and garlic extract treatment of H460 cells induced a rise in the level of Bax and a fall of Bcl-2 level. These results demonstrate that DAS, DADS and garlic extract are effective in reduction of anti-proliferative gene in NSCLC and suggest that modulation of apoptosis-associated cellular proteins by DAS, DADS and garlic extract may be the mechanism for apoptosis which merit further investigation as potential chemoprevention agents. PMID- 11048644 TI - Responsive site on the thrombospondin-1 promotor to down-regulation by phorbol 12 myristate 13-acetate in porcine aortic endothelial cells. AB - Thrombospondin-1 (TSP-1), a multifunctional extracellular matrix protein, inhibits neovascularization and is implicated in the regression of tumor growth and metastasis. We found that the synthesis of TSP-1 in porcine aortic endothelial (PAE) cells was decreased in a dose-dependent manner by phorbol 12 myristate 13-acetate (PMA) treatment in porcine aortic endothelial (PAE) cells. In this study, a responsive site on the TSP-1 promotor affected by PMA treatment in PAE was characterized. The level of TSP-1 mRNA was also decreased by PMA after 1 h and persisted that way for at least 24 h. PMA treatment and c-Jun overexpression suppressed the transcription of TSP-1 promotor-luciferase reporter gene. A deletion between -767 and -657 on the TSP-1 promotor neutralized the PMA induced down-regulation. In addition, oligo a (-767 approximately -723) was responsive to PMA-induced repression, while oligo b (-734 approximately -689) and c (-700 approximately -656) was not. Electrophoretic mobility shift assays showed that this PMA responsive element specifically bound a nuclear protein and that the binding activity was diminished by PMA treatment in PAE cells but not in Hep 3B cells. In supershift assay, potential regulatory elements in this region, SP1 and GATA-1, were not responsive to the inhibition of TSP-1 expression by PMA. Our results suggest that the repression of TSP-1 synthesis by PMA is mediated by blocking a particular unknown nuclear protein binding to the responsive site ( 767 approximately -735), which is regulated by c-Jun. PMID- 11048645 TI - An improved method for quantitative sugar analysis of glycoproteins. AB - Although there are numerous methods available to hydrolyze glycans utilizing strong acids, it all requires lengthy steps to obtain quantitative yield. We have developed a new simple one-step method for analysis of amino and neutral monosaccharides of glycoproteins quantitatively. Free monosaccharides were found to be stable during hydrolysis of glycans with 6 N HCI at 80 degrees C up to 2 h. Using this condition, analysis of free monosaccharides hydrolyzed from the bovine fetuin showed sugar composition of Gal: Man: GlcN: GaIN = 13.2: 11.0: 15.5: 2.6, which is closely matched with the reported value of 12.4: 9.6: 17.2: 2.7 (Townsend et al., ABRF News 8: 14, 1997). This method was shown to be applicable to varieties of well-characterized glycoproteins, erythropoietin, fibrinogen and soybean agglutinin. The amounts of sugars released under the condition were very close to the experimental values by other procedures or to the theoretical ones. This condition was found to be suitable for direct sugar analysis of fetuin, which have been immobilized onto polyvinylidene difluoride membrane. Based on these results, it support that the 6 N HCl/80 degrees C/2 h is the simplest method for quantitative analysis of monosaccharide composition of glycoproteins. PMID- 11048646 TI - Molecular characteristics of the inhibition of human neutrophil elastase by nonsteroidal antiinflammatory drugs. AB - Nonsteroidal antiinflammatory drugs(NSAIDs) are known as clinically effective agents for treatment of inflammatory diseases. Inhibition of cyclooxygenase has been thought to be a major facet of the pharmacological mechanism of NSAIDs. However, it is difficult to ascribe the antiinflammatory effects of NSAIDs solely to the inhibition of prostaglandin synthesis. Human neutrophil elastase (HNElastase; HNE, EC 3.4.21.37) has been known as a causative factor in inflammatory diseases. To investigate the specific relationship between HNElastase inhibition and specificity of molecular structure of several NSAIDs, HNElastase was purified by Ultrogel AcA54 gel filtration, CM-Sephadex ion exchange, and HPLC (with TSK 250 column) chromatography. HNElastase was inhibited by aspirin and salicylate in a competitive manner and by naproxen, ketoprofen, phenylbutazone, and oxyphenbutazone in a partial competative manner, but not by ibuprofen and tolmetin. HNElastase-phenylbutazone-complex showed strong Raman shifts at 200, 440, 1124, 1194, 1384, 1506, and 1768 cm(-1). The Raman bands 1194, 1384, and 1768 cm(-1) may represent evidences of the conformational change at -N=N-phi radical, pyrazol ring, and -C=O radical of the elastase-drug complex, respectively. Phenylbutazone might be bound to HNElastase by ionic and hydrophobic interaction, and masked the active site. Inhibition of HNElastase could be another mechanism of action of NSAIDs besides cyclooxygenase inhibition in the treatment of inflammatory diseases. Different inhibition characteristics of HNE-lastase by NSAIDs such as aspirin, phenylbutazone-like drugs and ineffective drugs could be important points for drawing the criteria for appropriate drugs in clinical application. PMID- 11048647 TI - Identification of Escherichia coli 8-oxoguanine endonuclease. AB - 7,8-Dihydro-8-oxoguanine (oh8Gua) endonuclease is a DNA repair enzyme in Escherichia coli to remove oh8Gua, a promutagenic DNA adduct. Due to the unique mode of enzyme action and substrate specificity, this DNA repair enzyme has been suggested to be identical to 2,6-diamino-4-hydroxyformamidopyrimidine (Fapy)-DNA glycosylase (Fpg). However, oh8Gua endonuclease had not been definitely identified because it had not been homogeneously purified. In this study, we attempted to purify and identify the enzyme. Through several purification procedures, we obtained two proteins (32 kD and 29 kD). The larger protein co migrated with Fpg in 12% SDS-PAGE gel. Sequences of N-terminal amino acids of these two proteins were identical to that of Fpg; the smaller one is a degraded product of oh8Gua endonuclease during purification steps. These results indicate that oh8Gua endonuclease is identical to Fpg, implying that oh8Gua in oxidatively damaged DNA rather than Fapy is more physiologically relevant substrate for Fpg. PMID- 11048648 TI - Effect of short-term ethanol on the proliferative response of Swiss 3T3 cells to mitogenic growth factors. AB - Both adaptive and deleterious responses of cells to ethanol are likely triggered by short-term interactions of the cells with ethanol. Many studies have demonstrated the direct effect of ethanol on growth factor-stimulated cell proliferation. Using Swiss 3T3 cells whose growth was inhibited by ethanol in a concentration-dependent manner, we further investigated the molecular mechanisms of acute ethanol treatment by examining its effect on EGF- and PDGF-mediated cellular signaling systems for the mitogenic function. Tyrosine autophosphorylation of the growth factor receptors was partially prevented by ethanol in intact cells. When ethanol was included before or after EGF stimulation, no effect on the receptor signaling was observed. Here we also report that ethanol inhibits activation of ERK induced by both EGF and PDGF. EGF induced JNK activation was reduced but PDGF-induced rapid JNK activation was delayed by the addition of ethanol. The balance between its inhibitory and stimulatory effect on the signaling molecules might determine the rate of cell growth. PMID- 11048649 TI - Dexamethasone enhances phospholipase D activity in M-1 cells. AB - Phospholipase D (PLD) is an enzyme involved in signal transduction and widely distributed in mammalian cells. The signal transduction pathways and role for phospholipid metabolism during hormonal response in cortical collecting duct remain partly undefined. It has been reported that dexamethasone increases transepithelial transport in M-1 cells that are derived from the mouse cortical collecting duct. We investigated the expression and activity of PLD in M-1 cells. Basal PLD activity of M-1 cells cultured in the presence of dexamethasone (5 microM) was higher than in the absence of dexamethasone. Dexamethasone and ATP activated PLD in M-1 cells but phorbol ester did not stimulate PLD activity. Vasopressin, bradykinin, dibutyryl cyclic AMP, and ionomycin were ineffective in activating PLD of the cells. The PLD2 isotype was detected by immunoprecipitation but PLD1 was not detected in M-1 cells. Addition of GTPgammaS and ADP ribosylation factor or phosphatidylinositiol 4,5-bisphosphate to digitonin permeabilized cells did not augment PLD activity. In intact cells PLD activity was increased by sodium oleate but there was no significant change between dexamethasone treated- and untreated cells by oleate. These results suggest that at least two types of PLD are present in M-1 cells and PLD plays a role in the corticosteroid-mediated response of cortical collecting duct cells. PMID- 11048650 TI - Sec-dependent protein export and the involvement of the molecular chaperone SecB. PMID- 11048651 TI - The hsp110 and Grp1 70 stress proteins: newly recognized relatives of the Hsp70s. AB - Both the Grp170 and Hsp110 families represent relatively conserved and distinct sets of stress proteins, within a more diverse category that also includes the Hsp70s. All of these families are found in a wide variety of organisms from yeasts to humans. Although Hsp110s or Grp170s are not Hsp70s any more than Hsp70s are Hsp110s or Grp170s, it is still reasonable to refer to this combination of related families as the Hsp70 superfamily based on arguments discussed above and since no obvious prokaryotic Hsp110 or Grp170 has yet been identified. These proteins are related to their counterparts in the Hsp70/Grp78 family of eukaryotic stress proteins but are characterized by significantly larger molecular weights. The members of the Grp170 family are characterized by C terminal ER retention sequences and are ER localized in yeasts and mammals. As a Grp, Grp170 is recognized to be coregulated with other major Grps by a well-known set of stress conditions, sometimes referred to as the unfolded protein response (Kozutsumi et al 1988; Nakaki et al 1989). The Hsp110 family members are localized in the nucleus and cytoplasm and, with other major Hsps, are also coregulated by a specific set of stress conditions, most notably including hyperthermic exposures. Hsp110 is sometimes called Hsp105, although it would be preferable to have a uniform term. The large Hsp70-like proteins are structurally similar to the Hsp70s but differ from them in important ways. In both the Grp170 and Hspl10 families, there is a long loop structure that is interposed between the peptide-binding ,-domain and the alpha-helical lid. In the Hsp110 family and Grp170, there are differing degrees of expansion in the alpha-helical domain and the addition of a C-terminal loop. This gives the appearance of much larger lid domains for Hsp110 and Grp170 compared with Hsp70. Both Hsp110 and Grp170 families have relatively conserved short sequences in the alpha-helical domain in the lid, which are conserved motifs in numerous proteins (we termed these motifs Magic and TedWylee as discussed earlier). The structural differences detailed in this review result in functional differences between the large (Grp170 and Hspl10) members of the Hsp70 superfamily, the most distinctive being an increased ability of these proteins to bind (hold) denatured polypeptides compared with Hsc70, perhaps related to the enlarged C-terminal helical domain. However, there is also a major difference between these large stress proteins; Hsp110 does not bind ATP in vitro, whereas Grp170 binds ATP avidly. The role of the Grp170 and Hsp110 stress proteins in cellular physiology is not well understood. Overexpression of Hsp110 in cultured mammalian cells increases thermal tolerance. Grp170 binds to secreted proteins in the ER and may be cooperatively involved in folding these proteins appropriately. These roles are similar to those of the Hsp70 family members, and, therefore, the question arises as to the differential roles played by the larger members of the superfamily. We have discussed evidence that the large members of the superfamily cooperate with members of the Hsp70 family, and these chaperones probably interact with a large number of chaperones and cochaperones in their functional activities. The fundamental point is that Hsp110 is found in conjunction with Hsp70 in the cytoplasm (and nucleus) and Grp170 is found in conjunction with78 in tha ER in every eucaryotic cell examined from yeast to humans. This would strongly argue that Hsp110 Grp170 exhibit functions in eucaryotes not effectively performed by Hsp70s or Grp78, respectively. Of interest in this respect is the observation that all Hsp110s loss of function or deletion mutants listed in the Drosophila deletion project database are lethal. The important task for the future is to determine the roles these conserved molecular chaperones play in normal and physiologically stressed cells. PMID- 11048652 TI - The distribution of heat shock proteins in the nervous system of the unstressed mouse embryo suggests a role in neuronal and non-neuronal differentiation. AB - Heat shock proteins (Hsps) act as molecular chaperones and are generally constitutively expressed in the absence of stress. Hsps are also inducible by a variety of stressors whose effects could be disastrous on the brain. It has been shown previously that Hsps are differentially expressed in glial and neuronal cells, as well as in the different structures of the brain. This differential expression has been related to specific functions distinct from their general chaperone function, such as intracellular transport. We investigated here the constitutive expression of 5 Hsps (the small Hsp, Hsp25, the constitutive Hsc70 and Hsp90beta, the mainly inducible Hsp70 and Hsp90alpha), and of a molecular chaperone, TCP-1alpha during mouse nervous system development. We analyzed, by immunohistochemistry, their distribution in the central nervous system and in the ganglia of the peripheral nervous system from day 9.5 (E9.5) to day 17.5 (E17.5) of gestation. Hsps are expressed in different cell classes (neuronal, glial, and vascular). The different proteins display different but often overlapping patterns of expression in different regions of the developing nervous system, suggesting unique roles at different stages of neural maturation. Their putative function in cell remodeling during migration or differentiation and in protein transport is discussed. Moreover we consider Hsp90 function in cell signaling and the role of Hsp25 in apoptosis protection. PMID- 11048653 TI - Immobilization stress induces c-Fos accumulation in liver. AB - Acute stress-induced injury in tissues has been revealed by both biochemical markers in plasma and microscopy. However, little is known of the mechanisms by which tissue integrity is restored. Recently, induction of early response genes such as c-fos has been reported in the heart and stomach of immobilized animals. Herein, we show that immobilization stress in mice increased plasma alanine aminotransferase activity, a marker of liver damage. c-Fos protein accumulation in liver was induced by stress after 20 minutes of immobilization and persisted for 3 hours. Immobilization also induced the release of epidermal growth factor (EGF) from submandibular salivary glands and a transient increase in EGF concentration in plasma. Although EGF administration induced a 2.5-fold increase in c-Fos mass in the liver of anesthetized mice, sialoadenectomy (which abolished the effect of immobilization on plasma EGF) did not affect the stress-induced rise in plasma alanine aminotransferase activity or liver c-Fos accumulation. Therefore, we conclude that immobilization stress induces c-Fos accumulation in liver and that this effect is not triggered by the increase in plasma EGF concentration. PMID- 11048654 TI - Induced hsp70 is in small, cytoplasmic complexes in a cell culture model of renal ischemia: a comparative study with heat shock. AB - A number of clinical conditions are known to result in the induction of heat shock proteins, but detailed studies on stress response have focused mostly on heat shock as a model. We have analyzed the induction and intracellular distribution of heat shock proteins in a reversible adenosine triphosphate (ATP) depletion model of renal ischemia. Two Hsp70 homologues, Hsp70 in the cytoplasm and BiP in the endoplasmic reticulum (ER) lumen, were found significantly induced during the recovery phase of ATP depletion. Other members of the heat shock protein family, such as Hsp90, constitutive Hsc70, and a related protein Hop60, were not induced. The induction of stress proteins on ATP depletion differed from that after heat shock in the kinds of proteins elaborated, their induction kinetics, and their intracellular distributions. Biochemical fractionation and indirect immunofluorescence experiments indicated that Hsp70 was predominantly cytoplasmic in the recovery phase of ischemia-like stress. Velocity sedimentation on sucrose gradients showed that induced Hsp70 sedimented as small, soluble complexes, ranging in size from 4S20,w to 8S20,w. The results suggest a role for induced Hsp70 that may be different from one of protecting aggregated proteins as under heat shock and emphasize the need for their characterization in other clinical conditions that result in stress response. PMID- 11048655 TI - Toxoplasma gondii Hsp70 as a danger signal in toxoplasma gondii-infected mice. AB - Toxoplasma gondii Hsp70, T gondii Hsp30/bag1, and surface antigen 1 messenger RNAs were shown to be useful in analyzing stage conversion of T gondii between bradyzoites and tachyzoites. The high-level expression of T gondii Hsp70 was correlated with mortality in interferon-gamma knockout mice infected with T gondii. Tgondii Hsp70 inhibited the induction of nitric oxide release by peritoneal macrophages of T gondii-infected mice. These findings identify T gondii Hsp70 as a danger signal during lethal, acute T gondii infection. PMID- 11048656 TI - Autoantibodies against chaperonin CCT in human sera with rheumatic autoimmune diseases: comparison with antibodies against other Hsp60 family proteins. AB - Chaperonin CCT containing t-complex polypeptide 1 is a cytosolic molecular chaperone that assists in the folding of actin, tubulin, and other proteins and is a member of the 60-kDa heat shock protein (Hsp60) family. We examined antibody titers against human CCT and other Hsp60 family members in the sera of patients with rheumatic autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematodes, Sjogren syndrome, and mixed connective tissue disease. Autoantibody titers against not only human mitochondrial Hsp60 but also CCT were significantly higher in the sera of patients with rheumatic autoimmune diseases than in healthy control sera. Although immunoglobulin G (IgG) titers against Escherichia coli GroEL were high in all the groups of sera tested, no significant differences in anti-GroEL responses were detected between patients and healthy controls. IgG titers against mycobacterial Hsp65 showed a similar pattern to titers of autoantibodies recognizing GroEL. Immunoabsorption experiments demonstrated that most of the autoantibodies recognizing CCT were cross-reactive with mitochondrial Hsp60, E coli GroEL, and mycobacterial Hsp65. Although most of the anti-Hsp60 IgG recognized CCT, anti-GroEL (or antimycobacterial Hsp65) IgG contained antibodies specific for GroEL (or mycobacterial Hsp65) in addition to antibodies cross-reactive with CCT and Hsp60. Results from immunoblot analyses, together with weak (15% to 20%) amino acid sequence identities between CCT and the other Hsp60 family members, suggested that CCT-reactive autoantibodies recognize conformational epitopes that are conserved among CCT and other Hsp60 family members. PMID- 11048657 TI - Analysis of the levels of conservation of the J domain among the various types of DnaJ-like proteins. AB - DnaJ-like proteins are defined by the presence of an approximately 73 amino acid region termed the J domain. This region bears similarity to the initial 73 amino acids of the Escherichia coli protein DnaJ. Although the structures of the J domains of E coli DnaJ and human heat shock protein 40 have been solved using nuclear magnetic resonance, no detailed analysis of the amino acid conservation among the J domains of the various DnaJ-like proteins has yet been attempted. A multiple alignment of 223 J domain sequences was performed, and the levels of amino acid conservation at each position were established. It was found that the levels of sequence conservation were particularly high in 'true' DnaJ homologues (ie, those that share full domain conservation with DnaJ) and decreased substantially in those J domains in DnaJ-like proteins that contained no additional similarity to DnaJ outside their J domain. Residues were also identified that could be important for stabilizing the J domain and for mediating the interaction with heat shock protein 70. PMID- 11048658 TI - The myocardial heat shock response following sodium salicylate treatment. AB - In cultured cells, salicylate has been shown to potentiate the induction of Hsp72 so that a mild heat stress (40 degrees C) in the presence of salicylate induces an Hsp72 response that is similar to a severe heat stress (42 degrees C). To determine whether salicylate can potentiate the myocardial Hsp70 response in vivo and confer protection from an ischemic stress, male Sprague-Dawley rats (250-300 g) were placed into 5 groups: (1) control, (2) salicylate only (400 mg/kg), (3) mild heat stress (40 degrees C for 15 minutes), (4) mild heat stress plus salicylate, and (5) severe heat stress (42 degrees C for 15 minutes). Twenty-four hours following salicylate treatment and/or heat stress, animals were anesthetized, their hearts rapidly isolated, and hemodynamic function evaluated using the Langendorff technique. Hsp72 content was subsequently assessed by Western blotting. Although salicylate in combination with a mild heat stress induced heat shock factor activation, only the hearts from severely heat-stressed animals (42 degrees C) demonstrated a significantly elevated myocardial Hsp72 content and a significantly enhanced postischemic recovery of left ventricular developed pressure and rates of contraction and relaxation. These results support the role for Hsp72 as a protective protein and suggest that neither salicylate treatment alone nor salicylate in combination with a mild heat stress potentiates the myocardial Hsp72 response. PMID- 11048659 TI - Chemotactic selection of Tetrahymena pyriformis GL induced with histamine, di iodotyrosine or insulin. AB - It has been hypothesized that in phylogeny the encounter between potential signalling molecules and the continously changing cell membrane could result in the formation of a ligand specific receptor. This chemical (hormonal) imprinting is then transmitted to the progeny generations. It is, however, very difficult to know whether the selection of cells with receptor-like patterns or amplification of complete receptor-like patterns led to the formation of the receptor-hormone complex. The new technique of 'chemotactic selection provides a physiological response-guided selection of cells. It also enables the testing of subpopulations with the characteristic selector ligand. We show here that of three chemotactic ligands (histamine, di-iodotyrosine (T2) and human insulin), insulin and T2 selected subpopulations express a significantly high chemotactic response. Since the control medium has a selector capacity itself, we introduced a chemotactic selection coefficient (Chsel) which facilitates the comparison of all groups. Using this factor we found that insulin (Chsel = 1.57), functions as a strong selector and T2 (Chsel = 0.98), was a weak selector. Morphometric evaluation of the cells showed a good correlation between chemotactic responsiveness and morphometric characteristics of subpopulations selected with insulin and histamine. T2 data suggest that the long lasting responsiveness is not general, but might be subpopulation specific. PMID- 11048660 TI - Beta-adrenergic receptor subtypes mediating lipolysis in porcine adipocytes. Studies with BRL-37344, a putative beta3-adrenergic agonist. AB - The beta3-selective adrenergic receptor ligand BRL 37344 (BRL) was used to differentiate the presence and functional role of beta-adrenergic receptor (betaAR) subtypes in pig tissues. BRL did not stimulate adenylyl cyclase in membrane preparations or increase lipolysis from pig adipocytes. In contrast to some species, BRL appears to be a poor agonist for the pig betaAR and is not a useful betaAR ligand. Based on displacement of [3H]dihydroalprenolol binding, BRL exhibited a 100-fold selectivity for pig betaAR subtypes in adipose and skeletal muscle membranes. The high affinity site was proposed to be the beta2AR. When used as an antagonist, BRL blockade of the high affinity site did not interfere with isoproterenol-stimulated lipolysis but did inhibit adenylyl cyclase activation. Results indicate that the high affinity betaAR (betaAR) is not linked to lipolysis, possibly due to intracellular compartmentalization. Therefore, betaAR subtypes may have function-specific effects. PMID- 11048661 TI - Dietary cadmium induces histopathological changes despite a sufficient metallothionein level in the liver and kidneys of the bank vole (Clethrionomys glareolus). AB - The objective of this study was to correlate hepatic and renal cadmium (Cd) accumulation, Cd-binding capacity of metallothionein (MT) and lipid peroxidation with the tissue injury in the male bank voles raised under short (8 h light/16 h dark) and long (16 h light/8 h dark) photoperiods that affect differently Cd accumulation and MT induction in these rodents. The animals were exposed to dietary Cd (0, 40 and 80 microg/g) for 6 weeks. The accumulation of Cd in the liver and kidneys appeared to be dose-dependent in bank voles from the two photoperiod groups; however, the short-photoperiod animals exhibited significantly higher concentrations of Cd in both organs than the long photoperiod bank voles. Cd-Binding capacity of MT in the liver and kidneys of bank voles from the long photoperiod was sufficiently high to bind and detoxify all Cd ions, while in the animals fed 80 microg Cd/g under the short photoperiod, the concentrations of Cd in both organs exceeded (by about 10 microg/g) the MT capacity. However, similar histopathological changes in the liver (a focal hepatocyte swelling and granuloma) and kidneys (a focal degeneration of proximal tubules) occurred in Cd-80 bank voles from the two photoperiods. Likewise, in either photoperiod group, dietary Cd brought about a similar, dose-dependent decrease in the hepatic and renal lipid peroxidation, which paralleled closely that of the iron (Fe) concentrations. These data indicate that: (1) MT does not protect the liver and kidneys against Cd-induced injury in the bank vole exposed to the higher level of dietary Cd; and (2) lipid peroxidation cannot be responsible for the tissue damage. It is hypothesized that dietary Cd produces histopathological changes indirectly, through depressing the tissue Fe and Fe dependent oxidative processes. PMID- 11048662 TI - The liver monooxygenase system of Brazilian freshwater fish. AB - Content of cytochromes b5 and P-450, and activities of NADPH-cytochrome c (P-450) reductase (NCR) and 7-ethoxyresorufin O-deethylase (EROD) were measured in liver microsomes prepared from two South American endemic fish, Brycon cephalus and Colossoma macropomum, from tilapia, Oreochromis niloticus, and from Swiss mice, Mus musculus, which served as a control. Strong hemoglobin binding to fish liver microsomal membranes (FLM) altered visible spectra of microsomal cytochromes. Consequently, special precautions during FLM preparation, including liver perfusion followed by repeated washing of microsomes, were required in the study of microsomal cytochromes from these fish. FLM from all fish studied here had a significantly lower content of microsomal cytochromes but a similar level of NCR and EROD activities compared to mouse liver microsomes (MLM). Strong response of the monooxygenase system in O. niloticus to water pollution was detected with both specific cytochrome P-450 content and EROD activity increasing sharply. The optical spectra of hemoglobin from B. cephalus and C. macropomum were analyzed and some differences in shape and relative extinction were observed compared to known hemoglobins. PMID- 11048663 TI - In vitro evaluation of newly synthesised [1,2,4]triazolo[1,5a]pyrimidine derivatives against Trypanosoma cruzi, Leishmania donovani and Phytomonas staheli. AB - The antiprotozoal activity of newly synthesised compounds, all [1,2,4]triazolo [1,5a]pyrimidine derivatives, was tested against the protozoan parasites Trypanosoma cruzi, Leishmania donovani and Phytotmonas staheli. Six of these compounds significantly inhibited in vitro cell growth of the epimastigote forms of T. cruzi, and the promastigote forms of L. donovani and P. staheli. Some of the compounds reached complete growth inhibition at 1 microg/ml for 48 h of parasite/drug interaction. None of the compounds tested showed significant toxicity against cells of Aedes albopictus, mouse macrophages J-774A.1 and Lycopersicum esculentum at dosages five times greater than used against parasites. PMID- 11048664 TI - Muscle and serum changes with salbutamol administration in aerobically exercised rats. AB - Treatment of experimental animals subjected to 90 days physical training programme plus repeated doses of salbutamol, a beta-adrenergic agonist, administered under two different regimes: therapeutic (16 microg/kg body weight, twice a day) and doping (3 mg/kg body weight, twice a day), caused a marked increase in size of skeletal (soleus, gastrocnemius and plantaris) leg muscles. Adrenergic involvement of salbutamol-linked hypertrophy was demonstrated by co administration of the non-specific beta-adrenergic antagonist D,L-propranolol (10 mg/kg body weight twice a day). The salbutamol-induced muscle hypertrophy was associated with an early increase in creatine phosphokinase (CK) and its myocardial isozyme (CKmb), without significant changes in lactate dehydrogenase (LDH), alanine aminotransferase (AAT) and aspartate aminotransferase (DAT). The induction of muscle-injury biomarkers was completely abolished by co administration of propranolol, thus suggesting the adrenergic involvement of these alterations. PMID- 11048665 TI - Cynomolgus monkey liver aldehyde oxidase: extremely high oxidase activity and an attempt at purification. AB - Aldehyde oxidase (EC 1.2.3.1) in monkey (Macaca fascicularis) liver was characterized. Liver cytosol exhibited extremely high benzaldehyde and phthalazine oxidase activities based on aldehyde oxidase, compared with those of rabbits, rats, mice and guinea pigs. Monkey liver aldehyde oxidase showed broad substrate specificity distinct from that of the enzyme from other mammals. Purified aldehyde oxidase from monkey liver cytosol showed two major bands and two minor bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS PAGE). These bands were also observed in Western blotting analysis using anti-rat aldehyde oxidase. The molecular mass of the enzyme was estimated to be 130-151 kDa by SDS-PAGE, and to be about 285 kDa by HPLC gel filtration. The results suggest that isoforms of aldehyde oxidase exist in monkey livers. PMID- 11048666 TI - Temporal and geographical variation in skeletal fluoride content of roe deer (Capreolus capreolus) from industrialized areas in Germany. AB - In order to study temporal and spatial variation of environmental fluoride levels, we analyzed the mandibular bone fluoride content of 157 roe deer (age range: 1-11 years) from two industrialized regions (Ruhr area: n = 76, sampling period 1955-1998; area W of Cologne: n = 81, sampling period 1983 1998) in the federal state of North Rhine-Westphalia, Germany. Bone fluoride values (dry weight basis) ranged between 150 mg F/kg (2 year-old specimen taken in 1997) and 5724 mg F/kg (10 year-old specimen taken in 1957). In both study areas, a pronounced decline in mandibular bone fluoride concentrations occurred over the respective sampling periods. In consequence, bone fluoride content of animals (both study areas pooled) taken during the period 1990-1998 was significantly (P < 0.00001) lower than that of roe deer from the period 1955 1989, while the two animal groups did not significantly differ in age. These findings are regarded as indicative of a considerable reduction of fluoride deposition into the animals' habitats, due to effective emission control measures. Bone fluoride values for the period 1990-1998 in the roe deer from the Ruhr area significantly (P < 0.005) exceeded those of the individuals from the study area W of Cologne, while the difference in age between the two groups was not significant. In both study areas, a significant (P < 0.00001) positive correlation between age and mandibular bone fluoride content (Ruhr area: rs = 0.601; area W of Cologne: rs = 0.725) was found for animals taken during this period. The present study underscores the suitability of analyzing skeletal fluoride concentrations in wild roe deer in order to monitor the magnitude of environmental contamination by fluoride and thereby to assess the efficiency of measures taken to reduce fluoride emissions from industrial sources. PMID- 11048667 TI - Variation in semicarbazide-sensitive amine oxidase activity in plasma and tissues of mammals. AB - Semicarbazide-sensitive amine oxidase (SSAO) (E.C. 1.4.3.6) is a group of enzymes with as yet poorly understood function which is widely present in nature. The variation in methodology for determination of activity, differences in substrates used and in nomenclature have made it difficult to compare SSAO in different species and tissues. Since SSAO is implicated in the pathophysiology of diabetes mellitus and congestive heart failure, our aim was to analyse the importance and abundance of SSAO in human plasma and tissues compared to other mammals. In plasma of ten different mammals, Vmax values were found to vary more than 10,000 fold, while KM differed much less; in human plasma SSAO activity is relatively low. In some species more than one SSAO entity was present in plasma. SSAO activity was ubiquitous in tissues of human, rat and pig, but varied considerably, both between species and between tissues. In human tissues, SSAO activity is higher than in tissues from rat and pig. Relative to monoamine oxidase-B there is also wide variation in SSAO, with much higher relative activities in human than in rat and pig tissues. We conclude that in plasma, SSAO activity is highest in ruminants, while in tissues, SSAO activity is more prominently present in human than in rat and pig. PMID- 11048668 TI - Acute effects of acephate and methamidophos on acetylcholinesterase activity, endocrine system and amino acid concentrations in rats. AB - Acute effects of acephate (Ace) and methamidophos (Met) on acetylcholinesterase activity, endocrine system and amino acid concentrations were studied in rats. The rats were injected intraperitoneally with Ace (500 mg/kg) or Met (5 mg/kg) and then sacrificed at 15 or 60 min after the injection (A15 and A60 for Ace and M15 and M60 for Met). The primary aim of this study was to determine whether the mammalian toxicity of Ace is solely due to its conversion to Met or the protection of Ace against Met-inhibited AChE is also an important factor. The second aim of this study was to study the effects of Ace and Met on the endocrine system and amino acid concentrations and whether or not these effects correlate with AChE inhibition and Met accumulation. The Ace or Met injected animals did not exhibit the signs of organophosphate (OP) poisoning within 15 min after the injection, but exhibited tremors at 45 min after the injection. Blood and brain AChE activity in A15 and M15 rats exhibited 55 to 75% inhibition while the enzyme activity in A60 and M60 rats exhibited 80 to 95% inhibition. Ace was metabolized to Met in rats both in vivo and in vitro. A 5 rats had significantly higher Met concentration in their liver, brain and adrenal glands compared to M 5 rats, and A60 rats had significantly higher Met concentrations in their blood, liver, brain and adrenal glands compared to M60 rats. Thus, tissue Met concentrations in Ace treated rats were significantly higher than in Met-treated rats and the inhibition of AChE activity was not consistent with the amount of metabolically formed Met, supporting the hypothesis that the Ace protection plays a role in the overall toxicity. Ace and Met both impaired circulating blood hormone and amino acid concentrations in rats. The endocrine effects of Ace and Met differed from their cholinergic effects, and were not proportional to the amount of Met present in different tissues obtained from the treatment groups. Plasma ACTH concentration was elevated in M60 rats but not in A60 rats. Thus, Ace may indirectly protect the pituitary against the toxic effects of Met. Unlike plasma ACTH levels, serum corticosterone and aldosterone levels were elevated in both A60 and M60 rats. Therefore, the effect of Met on the adrenal cortex may be mediated by the pituitary gland, while the effect of Ace may be due to direct Ace gland interaction. The decrease in the levels of some of the serum amino acids showed an increase in the energy demands in the treatment groups. PMID- 11048669 TI - Distribution of mycosporine-like amino acids in the sea hare Aplysia dactylomela: effect of diet on amounts and types sequestered over time in tissues and spawn. AB - We investigated the interaction of diet and accumulation of UV-absorbing mycosporine-like amino acids (MAAs) in body tissues and spawn of the sea hare Aplysia dactylomela to determine if MAA accumulation reflects type and level of dietary intake. Food sources were the red algae Acanthophora spicifera, Centroceras clavulatum, and Laurencia sp., and the green alga, Ulva lactuca. Adults were maintained on these foods for 40 days, after which feces were collected and tissues separated by dissection. Field animals were similarly sampled at this time. All spawn from experimental and field animals was collected over the study period. Samples, including seaweed foods, were analysed for six MAAs. Overnight consumption experiments using a variety of common seaweeds and one seagrass from A. dactylomela's habitat showed that the four seaweeds selected as foods were among those best-eaten by Aplysia. After 40 days levels of specific MAAs in the tissues of experimental animals showed excellent correlation with those in their diets, suggesting that the MAAs were dietarily-derived. Relative MAA contents in spawn from all diet groups correlated well with those in spawn from field animals. Commonest MAAs in spawn were porphyra-334, shinorine, and palythine, in this order. Concentrations of these MAAs were maintained at constant levels over time in spawn from all diet groups eating red algae and from field animals. Spawn from the Ulva dietary group showed an initial significant decline in MAA concentrations, but levels stabilized after the first 2 weeks. Skin was rich in porphyra-334 and shinorine, and levels of these in experimental animals correlated well with comparable levels in the skin of field animals. Digestive glands contained high levels of asterina-330, particularly those of the Centroceras dietary group, where concentrations reached a maximum of 21 mg dry g( 1). PMID- 11048670 TI - Inhibition of complement activation by water-soluble polysaccharides of some far eastern brown seaweeds. AB - Fucoidans and laminarans from Laminaria cichorioides, Laminaria japonica, Fucus evanescens, laminaran from Laminaria gurjanovae, other beta-D-glucans (translam, pustulan and zymosan) and lambda-carrageenan from Chondrus armatus were used to study the effect of water-soluble polysaccharides from seaweeds on the alternative pathway of complement (APC). beta-D-Glucans and fucoidans under study differed appreciably from each other by structural characteristics, and also by degree of purification. beta-D-glucans, on ability to bind complement, ranked in a line according to a degree of their purification. Highly purified beta-D glucans under study did not reveal an ability to bind complement. The fucoidans were divided conventionally into three groups according to their action on APC. Highly sulfated alpha-L-fucan from L. cichorioides with the greatest activity toward APC and caused 50% inhibition of reaction of activation (RA) of APC in a concentration of 0.5-0.7 mg/ml. Opposite 50% of inhibition of lysis of erythrocytes by sulfated heterogeneous fucoidan from L. japonica was achieved with 20 mg/ml. All other fucoidans and lambda-carrageenan have activity at 6-10 mg/ml concentration. Decreasing the sulfate content from 36% up to 9% in sample fucoidans under study was not reflected practically in the 50% inhibition concentration. Apparently, the degree of sulfating of fucoidans did not influence their action on APC. But the positive influence of fucose in structure of polysaccharide was obvious. PMID- 11048671 TI - Effect of three structurally related antimalarial drugs on liver microsomal components and lipid peroxidation in rats. AB - Changes in microsomal drug oxidizing enzymes, microsomal lipids, hepatic glutathione (GSH), glutathione S-trans-ferase (GST) and malondialdehyde (MDA) formation following administration of rats with therapeutic doses of three structurally related antimalarial drugs, amodiaquine (AQ), mefloquine (MQ) and halofantrine (HF) were investigated. There was a significant decrease in the activities of aniline hydroxylase, p-nitroanisole O-demethylase and pentoxyresorufin O-dealkylase in AQ, MQ and HF treated rats. AQ elicited the greatest effect with 50, 37 and 67% reductions in the activities of aniline hydroxylase, p-nitroanisole O-demethylase and pentoxyresorufin O-dealkylase, respectively. All the drugs prolonged hexobarbital-sleeping time to varying extents. The three drugs increased significantly the cholesterol per phospholipid ratio. AQ, MQ and HF decreased significantly the GSH level, GST activity and increased the formation of MDA. The results indicate that the alterations in hepatic microsomal components and lipid peroxidation caused by the antimalarials are related to the structural differences in the compounds. PMID- 11048673 TI - Further immunochemical and biocatalytic characterization of CYP1A1 from feral leaping mullet liver (Liza saliens) microsomes. AB - CYP1A is known to play important roles in the metabolism, detoxification and bioactivation of carcinogens and other xenobiotics in animals including fish. In our laboratory, CYP1A1 was obtained in a highly purified form with a specific content of 15-17 nmol P450 per mg protein from liver microsomes of feral fish, leaping mullet (Liza saliens). Purified mullet CYP1A1 showed a very high substrate specificities for 7-ethoxyresorufin and 7-methoxyresorufin in a reconstituted system containing purified fish P450 reductase and lipid. In addition, effects of each individual components of the reconstituted system, i.e., CYP1A1 and P450 reductase on 7-methoxyresorufin O-demethylase (MROD) activity were studied. 7-ethoxyresorufin O-deethylase (EROD) activity was strongly inhibited by alpha-naphthoflavone (ANF). At 0.5 and 2.5 microM. ANF inhibited EROD activity by 90 and 98%, respectively. Mullet CYP1A1 did not catalyze monooxygenations of other substrates such as aniline, ethylmorphine, N nitrosodimethylamine and p-nitrophenol. Antibodies produced against CYP1A1 orthologues in fish such as trout and scup showed strong cross-reactivity with the purified mullet CYP1A1. In addition, anti-L. saliens liver CYP1A1 produced in our laboratory inhibited both the EROD and MROD activities catalyzed by L. saliens liver microsomes but stronger inhibition was observed with EROD activity. On the other hand, anti-mullet CYP1A1 antibodies showed very weak cross reactivity with two proteins (presumably CYP1A1 and CYP1A2) in 3MC-treated rat liver microsomes. Moreover, 3MC-treated rat liver microsomal EROD activity was weakly inhibited by the anti-L. saliens liver CYP1A1. These results strongly suggested that the purified mullet CYP1A1 is structurally, functionally and immunochemically similar to the CYP1A1 homologues purified from other teleost species but functionally and immunochemically distinct from mammalian CYP1A1. PMID- 11048672 TI - The house musk shrew (Suncus murinus): a unique animal with extremely low level of expression of mRNAs for CYP3A and flavin-containing monooxygenase. AB - Expression of drug-metabolizing enzymes including cytochrome P450 (CYP) and flavin-containing monooxygenase (FMO) in various tissues of Suncus murinus (Suncus) were examined. Northern blot analysis showed that mRNAs hybridizable with cDNAs for rat CYP1A2, human CYP2A6, rat CYP2B1, human CYP2C8, human CYP2D6, rat CYP2E1, human CYP3A4 and rat CYP4A1 were expressed in various tissues from Suncus. The mRNA level of CYP2A in the Suncus lung was very high. Furthermore, it was found that the level of CYP2A mRNA in the Suncus lung was higher compared to the Suncus liver. The expression level of mRNA hybridizable with cDNA for human CYP3A4 was very low. The presence of CYP3A gene in Suncus was proven by the induction of the CYP with dexamethasone. Very low expression levels of mRNAs hybridizable with cDNAs for rat FMO1, rat FMO2, rat FMO3 and rat FMO5 were also seen in Suncus liver. No apparent hybridization band appeared when human FMO4 cDNA was used as a probe. The hepatic expression of mRNAs hybridizable with cDNAs for UDP-glucuronosyltransferase 1*6, aryl sulfotransferase, glutathione S transferase 1, carboxyesterase and microsomal epoxide hydrolase in the Suncus were observed. These results indicate that the Suncus is a unique animal species in that mRNAs for CYP3A and FMO are expressed at very low levels. PMID- 11048674 TI - Accumulation of protoporphyrin-IX (PpIX) in leukemic cell lines following induction by 5-aminolevulinic acid (ALA). AB - We investigated the amounts of protoporphyrin IX (PpIX) accumulated in noninduced cells and following 5-aminolevulinic acid (ALA)-induction. Following ALA administration PpIX increased in all leukemic cell lines under investigation (HEL 26-fold, HL60 6-fold, Jurkat 3-fold, ML2 2-fold) but not in lymphocytes. Compared to other cell lines studied, HEL cells showed the lowest basal level of PpIX and the largest relative increase in PpIX. Despite a high increase following ALA treatment, the PpIX level in HEL cells is almost as low as in lymphocytes. It is in agreement with their relatively low sensitivities of ALA-induced photodynamic therapy (ALA-PDT) shown previously [(Grebenova, D., Cajthamlova, H., Bartosova, J., Marinov, J., Klamova, H., Fuchs, O., Hrkal, Z., 1998. Selective destruction of leukemic cells by photo-activation of 5-aminolevulinic acid induced protoporphyrin IX. J. Photochem. Photobiol. B: Biol. 47, 74-81)]. The ferrochelatase activities in the individual cell lines are in good inverse correlation with PpIX amounts accumulated in the ALA-induced cells, but not with the relative increase (ratio) of PpIX levels from basal to ALA-induced ones. This is most apparent in HEL cells and lymphocytes. There is probably different regulation of heme biosynthesis in erythroid cells, which are therefore not suitable for the studies of ALA-PDT mechanism. PpIX was accumulated more extensively in absence of fetal calf serum than in its presence. The amounts of PpIX accumulated in cells decreased exponentially with increasing fetal calf serum concentration. PMID- 11048675 TI - A useful model to study the effect of high sugar concentrations upon growth and enzymic activities of toad embryos and larvae. AB - The aim of this study was to develop an oviparous model suitable for studying the differential effects and mechanisms by which a high concentration of extracellular glucose and other sugars produce diabetes complications, particularly body growth retardation during development. Hence, we studied the experimental conditions necessary to obtain measurable effects of high sugar concentrations (5-mM glucose, mannitol, fructose and galactose) upon body growth and development of Bufo arenarum embryos and larvae, and upon the activity of aspartate aminotransferase (AST), gamma-glutamyltransferase (GGT), and alkaline phosphatase (APP). Unfed animals kept in glucose showed lower body weight than controls at all stages, a condition only observed at stage 26 for animals kept in galactose and fructose. All animals reached the same stage of development regardless of the solution in which they were kept. Glucose and fructose significantly decreased the activity of all enzymes tested, while galactose only affected GGT activity. The model provides the first experimental evidence for the deleterious effect exerted in vivo by different sugars upon developing embryos and larvaes of Bufo arenarum. The results prove that this model might help to elucidate the effects and the pathogenic mechanisms of hyperglycemia upon growth and development of embryos exposed to environments with high sugar concentrations. It might also become a useful tool for testing the effectiveness of drugs designed to prevent the deleterious effect of such exposure. PMID- 11048676 TI - Whole-body metabolism of the organophosphorus pesticide, fenthion, in goldfish, Carassius auratus. AB - The in vivo metabolism of fenthion, an organophosphorus pesticide, and its sulfoxide (fenthion sulfoxide) was examined in goldfish (Carassius auratus). When goldfish were administered fenthion i.p. at a dose of 100 mg/kg, two metabolites were isolated from the tank water. They were identified as fenthion sulfoxide and fenthion oxon, in which > P = S of fenthion is transformed to > P = O, by comparing their mass and UV spectra, and their behavior in HPLC and TLC, with those of authentic standards. However, fenthion sulfone was not detected as a metabolite. The amounts of fenthion, fenthion sulfoxide and fenthion oxon excreted within 4 days were 2.7, 3.4 and 2.5%, of the initial dose of fenthion, respectively. Unchanged fenthion was detected in the body of the fish to the extent of 42-50% of the dose after 10 days, but fenthion sulfoxide and fenthion oxon showed very low concentrations. When fenthion sulfoxide was administered to the fish, about 70% of the dose was excreted unchanged into the tank water within 24 h, but little of the reduced compound, fenthion, was found. In contrast, fenthion was detected at 2.1% of dose in the body of goldfish as a metabolite of fenthion sulfoxide. The fact that fenthion is metabolized to the toxic oxon form in fish presumably has environmental and health implication for its use as a pesticide. PMID- 11048677 TI - In vitro metabolism of polychlorinated biphenyl congeners by beluga whale (Delphinapterus leucas) and pilot whale (Globicephala melas) and relationship to cytochrome P450 expression. AB - We measured rates of oxidative metabolism of two tetrachlorobiphenyl (TCB) congeners by hepatic microsomes of two marine mammal species, beluga whale and pilot whale, as related to content of selected cytochrome P450 (CYP) forms. Beluga liver microsomes oxidized 3,3',4,4'-TCB at rates averaging 21 and 5 pmol/min per mg for males and females, respectively, while pilot whale samples oxidized this congener at 0.3 pmol/min per mg or less. However, rates of 3,3',4,4'-TCB metabolism correlated with immunodetected CYP1A1 protein content in liver microsomes of both species. The CYP1A inhibitor alpha-naphthoflavone inhibited 3,3',4,4'-TCB metabolism by 40% in beluga, supporting a role for a cetacean CYP1A as a catalyst of this activity. Major metabolites of 3,3',4,4'-TCB generated by beluga liver microsomes were 4-OH-3,3',4',5-TCB and 5-OH-3,3',4,4' TCB (98% of total), similar to metabolites formed by other species CYP1A1, and suggesting a 4,5-epoxide-TCB intermediate. Liver microsomes of both species metabolized 2,2',5,5'-TCB at rates of 0.2-1.5 pmol/min per mg. Both species also expressed microsomal proteins cross-reactive with antibodies raised against some mammalian CYP2Bs (rabbit; dog), but not others (rat; scup). Whether CYP2B homologues occur and function in cetaceans is uncertain. This study demonstrates that PCBs are metabolized to aqueous-soluble products by cetacean liver enzymes, and that in beluga, rates of metabolism of 3,3',4,4'-TCB are substantially greater than those of 2,2',5,5'-TCB. These directly measured rates generally support the view that PCB metabolism plays a role in shaping the distribution patterns of PCB residues found in cetacean tissue. PMID- 11048678 TI - Metabolism of benzo(a)pyrene by duck liver microsomes. AB - The metabolism of benzo(a)pyrene [BP], a model carcinogenic PAH, by hepatic microsomes of two duck species, mallard (Anas platyrhynchos) and common merganser (Mergus merganser americanus) collected from chemically-contaminated and relatively non-contaminated areas was investigated. The rate of metabolism of BP by liver microsomes of common merganser and mallard collected from polluted areas (2,650 +/- 310 and 2,200 +/- 310 pmol/min per mg microsomal protein, respectively) was significantly higher than that obtained with liver microsomes of the two species collected from non-polluted areas (334 +/- 33 and 231 +/- 30 pmol/min per mg microsomal protein, respectively). The level of cytochrome P-450 1A1 was significantly higher in the liver microsomes of both duck species from the polluted areas as compared to the ducks from the non-polluted areas. The major BP metabolites, including BP-9, 10-diol, BP-4, 5-diol, BP-7, 8-diol, BP-1, 6-dione, BP-3, 6-dione, BP-6, 12-dione, 9-hydroxy-BP and 3-hydroxy-BP, formed by liver microsomes of both duck species from polluted and non-polluted areas, were qualitatively similar. However, the patterns of these metabolites were considerably different from each other. Liver microsomes of ducks from the polluted areas produced a higher proportion of benzo-ring dihydrodiols than the liver microsomes of ducks from the non-polluted areas, which converted a greater proportion of BP to BP-phenols. The predominant enantiomer of BP-7,8-diol formed by hepatic microsomes of the two duck species had an (-)R,R absolute stereochemistry. The data suggest that duck and rat liver microsomal enzymes have different regioselectivity but similar stereoselectivity in the metabolism of BP. PMID- 11048679 TI - Effect of age on lipid peroxides, lipofuscin and ascorbic acid contents of the lungs of male garden lizard. AB - Oxidative damage was assessed through the estimation of lipid peroxides (LP) in the lungs of an ageing short-lived species of reptile, Calotes versicolor, commonly known as the garden lizard. Attempts were also made to trace its relationship with the age pigment, lipofuscin and the antioxidant ascorbic acid. While LP increased with advancing age the contents of both lipofuscin and ascorbic acid did not show appreciable change during maturation ( < 1-1 year old) but declined during senescence phase (1 to 2-4 year old). While the pattern of age associated changes in LP and ascorbic acid indicate similarity with the pattern observed in most of the mammals, the reduction of lipofuscin in older lizards is a significant departure from the common trend. PMID- 11048680 TI - Growth hormone inhibits growth hormone secretion from the rainbow trout pituitary in vitro. AB - Growth hormone (GH) secretion in salmonids and other fish is under the control of a number of hypothalamic factors, but negative feed-back regulation by circulating hormones can also be of importance for the regulation of GH secretion. Mammalian studies show that GH has a negative feed-back effect on its own secretion. In order to elucidate if GH levels present a direct ultra-short negative feedback loop at the pituitary level GH secretion was studied in intact pituitaries from 50 g fish in an in vitro perifusion system. Following an initial equilibrium period pituitaries were exposed to five increasing concentrations (1 1,000 ng ml(-1)) of ovine GH (oGH) in 20-min steps, before being returned to a GH free perifusion. Ovine GH caused a significant dose-dependent inhibition of GH secretion and it is concluded that GH can exert a direct negative feedback control on GH secretion at the pituitary level. PMID- 11048681 TI - Molecular characterization and developmental expression of the aryl hydrocarbon receptor from the chick embryo. AB - The aryl hydrocarbon receptor (AhR) was cloned from the chick embryo and its function and developmental expression characterized. Chicken AhR cDNA coded for 858 amino acid protein and 396 bp of 3' UTR. The basic helix loop helix domain exhibited 87-100% amino acid identity to avian, mammalian, and amphibian AhR, and 69-74% to piscine AhR. The PAS (Per-ARNT-Sim) region was slightly less well conserved with (a) 97% identity to other avian sequences, (b) 81-86% to amphibian and mammalian AhR, and (c) 64-69% with piscine AhR. The carboxy terminus diverged the most among species with less than 53% amino acid identity between chicken and any available mammalian and piscine AhR sequences. The chicken AhR RNA and protein were 6.1 kb and 103 kDa, respectively. Chicken AhR dimerized with human AhR nuclear translocator and bound the mammalian dioxin-response element in a ligand-dependent manner. AhR protein was detected in neural ganglia; smooth, cardiac, and skeletal muscle; and epithelium involved in epithelial-to mesenchymal transformations, such as pituitary, gastrointestinal tract, limb apical-ectodermal ridge, and kidney collecting ducts. AhR mRNA was detected in all tissues expressing protein, except myocardium. Cytochrome P4501A4 mRNA was highly induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in a subset of tissues expressing AhR, including small intestine, liver, kidney, blood vessels, and outflow tract myocardium. In conclusion, the AhR sequence and function is highly conserved between birds and mammals, and although many tissues express AhR during chick embryo development, only a subset are responsive to TCDD induction of CYP1A4. PMID- 11048682 TI - Binding characteristics of [3H]17beta-estradiol in the hypothalamus of juvenile rainbow trout, Oncorhynchus mykiss. AB - Gonadal steroids in the salmonid brain, acting through cellular receptors, may be responsible for the modulation of neuronal activity and organization of reproductive behaviors. We report our findings on the use of [3H]17beta-estradiol (E2) to identify intracellular estrogen receptors (ERs) in the hypothalamus of juvenile rainbow trout, Oncorhynchus mykiss. Specific binding (B(SP)) of [3H]E2 was tissue dependent between 0.5 and 2.25 hypothalamus equivalents for cytosol and nuclear extract preparations, respectively. B(SP) in cytosol fractions increased with time and reached maximum levels (4.18 nM) at 2.5 h incubation; by contrast, B(SP) in nuclear extract increased with time to achieve maximum levels (3.9 nM) by 2 h incubation. The association rate constants (k(+1)) for cytosol and nuclear extract preparations were 1.10 +/- 0.02 x 10(8) M(-1) min(-1) and 1.27 +/- 0.04 x 10(8) M(-1) min(-1), respectively. Equilibrium bound B(SP) dissociated from cytosol preparations with a half life (t1-2) of 42 min and a dissociation rate constant (k(-1)) of 1.01 +/- 0.03 min(-1). B(SP) dissociated from nuclear extract preparations with a t1-2 = 45 min and k(-1)= 0.92 +/- 0.01 min(-1) x B(SP) was saturable in both extract preparations with a calculated equilibrium dissociation constant (Kd) of 1.46 +/- 0.1 nM (cytosol) and 2.37 +/- 0.2 nM (nuclear), and a maximum number of binding sites (B(MAX)) of 50.85 +/- 3.2 fmol mg(-1) protein and 61.74 +/- 2.65 fmol mg(-1) protein, respectively. In both preparations, B(SP) was differentially displaced by structurally similar compounds with a rank order of potency of E2 > estrone > estriol > 17alpha ethynyl estradiol > testosterone >> progesterone = tamoxifen >> cortisol > dexamethasone >> > beta-sitosterol. These properties of specifically bound [3H]E2 suggest the presence of an ER in the hypothalamus of juvenile rainbow trout comparable with ERs identified in salmonid liver. PMID- 11048684 TI - Blurred budget is bad news. PMID- 11048683 TI - In defence of animal research. PMID- 11048686 TI - Fathers of electronic revolution are rewarded. PMID- 11048685 TI - Medicine Nobel goes to raiders of the brain's chemical secrets. PMID- 11048687 TI - Plastics that conduct win inventors chemistry prize. PMID- 11048688 TI - Gore and Bush back rise in science spending. PMID- 11048689 TI - Project offers free mouse sequence. PMID- 11048690 TI - Anger as Spain boosts R&D figures with defence money. PMID- 11048691 TI - Astrometry mission wins German approval. PMID- 11048692 TI - Ig Nobel glory for levitating frogs and collapsing toilets. PMID- 11048693 TI - Science for art's sake. PMID- 11048694 TI - Poorly conducted (or reported) animal tests put humans at risk. PMID- 11048695 TI - Why don't creationists use private schools? PMID- 11048696 TI - Survival on the edge: the tube worm's strategy. PMID- 11048698 TI - Win a Nobel prize! PMID- 11048697 TI - A victim of truth. PMID- 11048699 TI - Electrochemistry. Making a potential difference. PMID- 11048700 TI - Computational neuroscience. Building blocks of movement. PMID- 11048701 TI - Conservation biology. Seeds of doubt. PMID- 11048702 TI - Stirring times in the Southern Ocean. PMID- 11048703 TI - Guarding against mutation. PMID- 11048704 TI - Evolutionary biology. Use it or lose it. PMID- 11048705 TI - Before striking gold in gold-ruby glass. PMID- 11048706 TI - Facultative worker policing in a wasp. PMID- 11048707 TI - Maternal effect of Hsf1 on reproductive success. PMID- 11048708 TI - Coexistence and resource competition. PMID- 11048709 TI - A mesoscale phytoplankton bloom in the polar Southern Ocean stimulated by iron fertilization. AB - Changes in iron supply to oceanic plankton are thought to have a significant effect on concentrations of atmospheric carbon dioxide by altering rates of carbon sequestration, a theory known as the 'iron hypothesis'. For this reason, it is important to understand the response of pelagic biota to increased iron supply. Here we report the results of a mesoscale iron fertilization experiment in the polar Southern Ocean, where the potential to sequester iron-elevated algal carbon is probably greatest. Increased iron supply led to elevated phytoplankton biomass and rates of photosynthesis in surface waters, causing a large drawdown of carbon dioxide and macronutrients, and elevated dimethyl sulphide levels after 13 days. This drawdown was mostly due to the proliferation of diatom stocks. But downward export of biogenic carbon was not increased. Moreover, satellite observations of this massive bloom 30 days later, suggest that a sufficient proportion of the added iron was retained in surface waters. Our findings demonstrate that iron supply controls phytoplankton growth and community composition during summer in these polar Southern Ocean waters, but the fate of algal carbon remains unknown and depends on the interplay between the processes controlling export, remineralisation and timescales of water mass subduction. PMID- 11048710 TI - Crystal structures of mismatch repair protein MutS and its complex with a substrate DNA. AB - DNA mismatch repair is critical for increasing replication fidelity in organisms ranging from bacteria to humans. MutS protein, a member of the ABC ATPase superfamily, recognizes mispaired and unpaired bases in duplex DNA and initiates mismatch repair. Mutations in human MutS genes cause a predisposition to hereditary nonpolyposis colorectal cancer as well as sporadic tumours. Here we report the crystal structures of a MutS protein and a complex of MutS with a heteroduplex DNA containing an unpaired base. The structures reveal the general architecture of members of the MutS family, an induced-fit mechanism of recognition between four domains of a MutS dimer and a heteroduplex kinked at the mismatch, a composite ATPase active site composed of residues from both MutS subunits, and a transmitter region connecting the mismatch-binding and ATPase domains. The crystal structures also provide a molecular framework for understanding hereditary nonpolyposis colorectal cancer mutations and for postulating testable roles of MutS. PMID- 11048712 TI - Dynamics of singularities in a constrained elastic plate. AB - Large deformations of thin elastic plates usually lead to the formation of singular structures which are either linear (ridges) or pointlike (developable cones). These structures are thought to be generic for crumpled plates, although they have been investigated quantitatively only in simplified geometries. Previous studies have also shown that a large number of singularities are generated by successive instabilities. Here we study, experimentally and numerically, a generic situation in which a plate is initially bent in one direction into a cylindrical arch, then deformed in the other direction by a load applied at its centre. This induces the generation of pairs of singularities; we study their position, their dynamics and the corresponding resistance of the plate to deformation. We solve numerically the equations describing large deformations of plates; developable cones are predicted, in quantitative agreement with the experiments. We use geometrical arguments to predict the observed patterns, assuming that the energy of the plate is given by the energy of the singularities. PMID- 11048711 TI - The crystal structure of DNA mismatch repair protein MutS binding to a G x T mismatch. AB - DNA mismatch repair ensures genomic integrity on DNA replication. Recognition of a DNA mismatch by a dimeric MutS protein initiates a cascade of reactions and results in repair of the newly synthesized strand; however, details of the molecular mechanism remain controversial. Here we present the crystal structure at 2.2 A of MutS from Escherichia coli bound to a G x T mismatch. The two MutS monomers have different conformations and form a heterodimer at the structural level. Only one monomer recognizes the mismatch specifically and has ADP bound. Mismatch recognition occurs by extensive minor groove interactions causing unusual base pairing and kinking of the DNA. Nonspecific major groove DNA-binding domains from both monomers embrace the DNA in a clamp-like structure. The interleaved nucleotide-binding sites are located far from the DNA. Mutations in human MutS alpha (MSH2/MSH6) that lead to hereditary predisposition for cancer, such as hereditary non-polyposis colorectal cancer, can be mapped to this crystal structure. PMID- 11048713 TI - Interconversion of single and double helices formed from synthetic molecular strands. AB - Synthetic single-helical conformations are quite common, but the formation of double helices based on recognition between the two constituent strands is relatively rare. Known examples include duplex formation through base-pair specific hydrogen bonding and stacking, as found in nucleic acids and their analogues, and polypeptides composed of amino acids with alternating L and D configurations. Some synthetic polymers and self-assembled fibres have double helical winding induced by van der Waals interactions. A third mode of non covalent interaction, coordination of organic ligands to metal ions, can give rise to double, triple and quadruple helices, although in this case the assembly is driven by the coordination geometry of the metal and the structure of the ligands, rather than by direct inter-strand complementarity. Here we describe a family of oligomeric molecules with bent conformations, which exhibit dynamic exchange between single and double molecular helices in solution, through spiral sliding of the synthetic oligomer strands. The bent conformations leading to the helical shape of the molecules result from intramolecular hydrogen bonding within 2'-pyridyl-2-pyridinecarboxamide units, with extensive intermolecular aromatic stacking stabilizing the double-stranded helices that form through dimerization. PMID- 11048714 TI - Prototype systems for rechargeable magnesium batteries. AB - The thermodynamic properties of magnesium make it a natural choice for use as an anode material in rechargeable batteries, because it may provide a considerably higher energy density than the commonly used lead-acid and nickel-cadmium systems. Moreover, in contrast to lead and cadmium, magnesium is inexpensive, environmentally friendly and safe to handle. But the development of Mg batteries has been hindered by two problems. First, owing to the chemical activity of Mg, only solutions that neither donate nor accept protons are suitable as electrolytes; but most of these solutions allow the growth of passivating surface films, which inhibit any electrochemical reaction. Second, the choice of cathode materials has been limited by the difficulty of intercalating Mg ions in many hosts. Following previous studies of the electrochemistry of Mg electrodes in various non-aqueous solutions, and of a variety of intercalation electrodes, we have now developed rechargeable Mg battery systems that show promise for applications. The systems comprise electrolyte solutions based on Mg organohaloaluminate salts, and Mg(x)Mo3S4 cathodes, into which Mg ions can be intercalated reversibly, and with relatively fast kinetics. We expect that further improvements in the energy density will make these batteries a viable alternative to existing systems. PMID- 11048715 TI - Importance of stirring in the development of an iron-fertilized phytoplankton bloom. AB - The growth of populations is known to be influenced by dispersal, which has often been described as purely diffusive. In the open ocean, however, the tendrils and filaments of phytoplankton populations provide evidence for dispersal by stirring. Despite the apparent importance of horizontal stirring for plankton ecology, this process remains poorly characterized. Here we investigate the development of a discrete phytoplankton bloom, which was initiated by the iron fertilization of a patch of water (7 km in diameter) in the Southern Ocean. Satellite images show a striking, 150-km-long bloom near the experimental site, six weeks after the initial fertilization. We argue that the ribbon-like bloom was produced from the fertilized patch through stirring, growth and diffusion, and we derive an estimate of the stirring rate. In this case, stirring acts as an important control on bloom development, mixing phytoplankton and iron out of the patch, but also entraining silicate. This may have prevented the onset of silicate limitation, and so allowed the bloom to continue for as long as there was sufficient iron. Stirring in the ocean is likely to be variable, so blooms that are initially similar may develop very differently. PMID- 11048716 TI - Effect of iron supply on Southern Ocean CO2 uptake and implications for glacial atmospheric CO2. AB - Photosynthesis by marine phytoplankton in the Southern Ocean, and the associated uptake of carbon, is thought to be currently limited by the availability of iron. One implication of this limitation is that a larger iron supply to the region in glacial times could have stimulated algal photosynthesis, leading to lower concentrations of atmospheric CO2. Similarly, it has been proposed that artificial iron fertilization of the oceans might increase future carbon sequestration. Here we report data from a whole-ecosystem test of the iron limitation hypothesis in the Southern Ocean, which show that surface uptake of atmospheric CO2 and uptake ratios of silica to carbon by phytoplankton were strongly influenced by nanomolar increases of iron concentration. We use these results to inform a model of global carbon and ocean nutrients, forced with atmospheric iron fluxes to the region derived from the Vostok ice-core dust record. During glacial periods, predicted magnitudes and timings of atmospheric CO2 changes match ice-core records well. At glacial terminations, the model suggests that forcing of Southern Ocean biota by iron caused the initial approximately 40 p.p.m. of glacial-interglacial CO2 change, but other mechanisms must have accounted for the remaining 40 p.p.m. increase. The experiment also confirms that modest sequestration of atmospheric CO2 by artificial additions of iron to the Southern Ocean is in principle possible, although the period and geographical extent over which sequestration would be effective remain poorly known. PMID- 11048717 TI - The possible subduction of continental material to depths greater than 200 km. AB - Determining the depth to which continental lithosphere can be subducted into the mantle at convergent plate boundaries is of importance for understanding the long term growth of supercontinents as well as the dynamic processes that shape such margins. Recent discoveries of coesite and diamond in regional ultrahigh-pressure (UHP) metamorphic rocks has demonstrated that continental material can be subducted to depths of at least 120 km (ref. 1), and subduction to depths of 150 300 km has been inferred from garnet peridotites in orogenic UHP belts based on several indirect observations. But continental subduction to such depths is difficult to trace directly in natural UHP metamorphic crustal rocks by conventional mineralogical and petrological methods because of extensive late stage recrystallization and the lack of a suitable pressure indicator. It has been predicted from experimental work, however, that solid-state dissolution of pyroxene should occur in garnet at depths greater than 150 km (refs 6-8). Here we report the observation of high concentrations of clinopyroxene, rutile and apatite exsolutions in garnet within eclogites from Yangkou in the Sulu UHP metamorphic belt, China. We interpret these data as resulting from the high pressure formation of pyroxene solid solutions in subducted continental material. Appropriate conditions for the Na2O concentrations and octahedral silicon observed in these samples are met at depths greater than 200 km. PMID- 11048718 TI - The population genetics of ecological specialization in evolving Escherichia coli populations. AB - When organisms adapt genetically to one environment, they may lose fitness in other environments. Two distinct population genetic processes can produce ecological specialization-mutation accumulation and antagonistic pleiotropy. In mutation accumulation, mutations become fixed by genetic drift in genes that are not maintained by selection; adaptation to one environment and loss of adaptation to another are caused by different mutations. Antagonistic pleiotropy arises from trade-offs, such that the same mutations that are beneficial in one environment are detrimental in another. In general, it is difficult to distinguish between these processes. We analysed the decay of unused catabolic functions in 12 lines of Escherichia coli propagated on glucose for 20,000 generations. During that time, several lines evolved high mutation rates. If mutation accumulation is important, their unused functions should decay more than the other lines, but no significant difference was observed. Moreover, most catabolic losses occurred early in the experiment when beneficial mutations were being rapidly fixed, a pattern predicted by antagonistic pleiotropy. Thus, antagonistic pleiotropy appears more important than mutation accumulation for the decay of unused catabolic functions in these populations. PMID- 11048719 TI - Natural selection and sympatric divergence in the apple maggot Rhagoletis pomonella. AB - In On the Origin of Species, Darwin proposed that natural selection had a fundamental role in speciation. But this view receded during the Modern Synthesis when allopatric (geographic) models of speciation were integrated with genetic studies of hybrid sterility and inviability. The sympatric hypothesis posits that ecological specialization after a host shift can result in speciation in the absence of complete geographic isolation. The apple maggot, Rhagoletis pomonella, is a model for sympatric speciation in progress. Hawthorn (Crataegus spp.) is the native host for R. pomonella in N. Americas. But in the mid-1800s, a new population formed on introduced, domesticated apple (Malus pumila). Recent studies have conferred 'host race' status on apple flies as a potentially incipient species, partially isolated from haw flies owing to host-related adaptation. However, the source of selection that differentiates apple and haw flies is unresolved. Here we document a gene-environment interaction (fitness trade-off) that is related to host phenology and that genetically differentiates the races. PMID- 11048720 TI - Learning of action through adaptive combination of motor primitives. AB - Understanding how the brain constructs movements remains a fundamental challenge in neuroscience. The brain may control complex movements through flexible combination of motor primitives, where each primitive is an element of computation in the sensorimotor map that transforms desired limb trajectories into motor commands. Theoretical studies have shown that a system's ability to learn action depends on the shape of its primitives. Using a time-series analysis of error patterns, here we show that humans learn the dynamics of reaching movements through a flexible combination of primitives that have gaussian-like tuning functions encoding hand velocity. The wide tuning of the inferred primitives predicts limitations on the brain's ability to represent viscous dynamics. We find close agreement between the predicted limitations and the subjects' adaptation to new force fields. The mathematical properties of the derived primitives resemble the tuning curves of Purkinje cells in the cerebellum. The activity of these cells may encode primitives that underlie the learning of dynamics. PMID- 11048721 TI - Netrin-1-mediated axon outgrowth and cAMP production requires interaction with adenosine A2b receptor. AB - The netrins, a family of laminin-related secreted proteins, are critical in controlling axon elongation and pathfinding. The DCC (for deleted in colorectal cancer) protein was proposed as a receptor for netrin-1 in the light of many observations including the inhibition of netrin-1-mediated axon outgrowth and attraction in the presence of an anti-DCC antiserum, the similitude of nervous system defects in DCC and netrin-1 knockout mice and the results of receptor swapping experiments. Previous studies have failed to show a direct interaction of DCC with netrin-1 (ref. 10), suggesting the possibility of an additional receptor or co-receptor. Here we show that DCC interacts with the membrane associated adenosine A2b receptor, a G-protein-coupled receptor that induces cAMP accumulation on binding adenosine. We show that A2b is actually a netrin-1 receptor and induces cAMP accumulation on binding netrin-1. Finally, we show that netrin-1-dependent outgrowth of dorsal spinal cord axons directly involves A2b. Together our results indicate that the growth-promoting function of netrin-1 may require a receptor complex containing DCC and A2b. PMID- 11048722 TI - Somatic support cells restrict germline stem cell self-renewal and promote differentiation. AB - Stem cells maintain populations of highly differentiated, short-lived cell-types, including blood, skin and sperm, throughout adult life. Understanding the mechanisms that regulate stem cell behaviour is crucial for realizing their potential in regenerative medicine. A fundamental characteristic of stem cells is their capacity for asymmetric division: daughter cells either retain stem cell identity or initiate differentiation. However, stem cells are also capable of symmetric division where both daughters remain stem cells, indicating that mechanisms must exist to balance self-renewal capacity with differentiation. Here we present evidence that support cells surrounding the stem cells restrict self renewal and control stem cell number by ensuring asymmetric division. Loss of function of the Drosophila Epidermal growth factor receptor in somatic cells disrupted the balance of self-renewal versus differentiation in the male germline, increasing the number of germline stem cells. We propose that activation of this receptor specifies normal behaviour of somatic support cells; in turn, the somatic cells play a guardian role, providing information that prevents self-renewal of stem cell identity by the germ cell they enclose. PMID- 11048723 TI - Somatic control over the germline stem cell lineage during Drosophila spermatogenesis. AB - Stem cells divide both to produce new stem cells and to generate daughter cells that can differentiate. The underlying mechanisms are not well understood, but conceptually are of two kinds. Intrinsic mechanisms may control the unequal partitioning of determinants leading to asymmetric cell divisions that yield one stem cell and one differentiated daughter cell. Alternatively, extrinsic mechanisms, involving stromal cell signals, could cause daughter cells that remain in their proper niche to stay stem cells, whereas daughter cells that leave this niche differentiate. Here we use Drosophila spermatogenesis as a model stem cell system to show that there are excess stem cells and gonialblasts in testes that are deficient for Raf activity. In addition, the germline stem cell population remains active for a longer fraction of lifespan than in wild type. Finally, raf is required in somatic cells that surround germ cells. We conclude that a cell-extrinsic mechanism regulates germline stem cell behaviour. PMID- 11048724 TI - The complete sequence of the mucosal pathogen Ureaplasma urealyticum. AB - The comparison of the genomes of two very closely related human mucosal pathogens, Mycoplasma genitalium and Mycoplasma pneumoniae, has helped define the essential functions of a self-replicating minimal cell, as well as what constitutes a mycoplasma. Here we report the complete sequence of a more distant phylogenetic relative of those bacteria, Ureaplasma urealyticum (parvum biovar), which is also a mucosal pathogen of humans. It is the third mycoplasma to be sequenced, and has the smallest sequenced prokaryotic genome except for M. genitalium. Although the U. urealyticum genome is similar to the two sequenced mycoplasma genomes, features make this organism unique among mycoplasmas and all bacteria. Almost all ATP synthesis is the result of urea hydrolysis, which generates an energy-producing electrochemical gradient. Some highly conserved eubacterial enzymes appear not to be encoded by U. urealyticum, including the cell-division protein FtsZ, chaperonins GroES and GroEL, and ribonucleoside diphosphate reductase. U. urealyticum has six closely related iron transporters, which apparently arose through gene duplication, suggesting that it has a kind of respiration system not present in other small genome bacteria The genome is only 25.5% G+C in nucleotide content, and the G+C content of individual genes may predict how essential those genes are to ureaplasma survival. PMID- 11048725 TI - Quorum-sensing signals indicate that cystic fibrosis lungs are infected with bacterial biofilms. AB - The bacterium Pseudomonas aeruginosa permanently colonizes cystic fibrosis lungs despite aggressive antibiotic treatment. This suggests that P. aeruginosa might exist as biofilms--structured communities of bacteria encased in a self-produced polymeric matrix--in the cystic fibrosis lung. Consistent with this hypothesis, microscopy of cystic fibrosis sputum shows that P. aeruginosa are in biofilm-like structures. P. aeruginosa uses extracellular quorum-sensing signals (extracellular chemical signals that cue cell-density-dependent gene expression) to coordinate biofilm formation. Here we found that cystic fibrosis sputum produces the two principal P. aeruginosa quorum-sensing signals; however, the relative abundance of these signals was opposite to that of the standard P. aeruginosa strain PAO1 in laboratory broth culture. When P. aeruginosa sputum isolates were grown in broth, some showed quorum-sensing signal ratios like those of the laboratory strain. When we grew these isolates and PAO1 in a laboratory biofilm model, the signal ratios were like those in cystic fibrosis sputum. Our data support the hypothesis that P. aeruginosa are in a biofilm in cystic fibrosis sputum. Moreover, quorum-sensing signal profiling of specific P. aeruginosa strains may serve as a biomarker in screens to identify agents that interfere with biofilm development. PMID- 11048726 TI - Messenger RNA targeting of rice seed storage proteins to specific ER subdomains. AB - Rice seeds, a rich reserve of starch and protein, are a major food source in many countries. Unlike the seeds of other plants, which typically accumulate one major type of storage protein, rice seeds use two major classes, prolamines and globulin-like glutelins. Both storage proteins are synthesized on the endoplasmic reticulum (ER) and translocated to the ER lumen, but are then sorted into separate intracellular compartments. Prolamines are retained in the ER lumen as protein bodies whereas glutelins are transported and stored in protein storage vacuoles. Mechanisms responsible for the retention of prolamines within the ER lumen and their assembly into intracisternal inclusion granules are unknown, but the involvement of RNA localization has been suggested. Here we show that the storage protein RNAs are localized to distinct ER membranes and that prolamine RNAs are targeted to the prolamine protein bodies by a mechanism based on RNA signal(s), a process that also requires a translation initiation codon. Our results indicate that the ER may be composed of subdomains that specialize in the synthesis of proteins directed to different compartments of the plant endomembrane system. PMID- 11048727 TI - The biochemistry of apoptosis. AB - Apoptosis--the regulated destruction of a cell--is a complicated process. The decision to die cannot be taken lightly, and the activity of many genes influence a cell's likelihood of activating its self-destruction programme. Once the decision is taken, proper execution of the apoptotic programme requires the coordinated activation and execution of multiple subprogrammes. Here I review the basic components of the death machinery, describe how they interact to regulate apoptosis in a coordinated manner, and discuss the main pathways that are used to activate cell death. PMID- 11048728 TI - Defying death after DNA damage. AB - DNA damage frequently triggers death by apoptosis. The irreversible decision to die can be facilitated or forestalled through integration of a wide variety of stimuli from within and around the cell. Here we address some fundamental questions that arise from this model. Why should DNA damage initiate apoptosis in the first place? In damaged cells, what are the alternatives to death and why should they be selected in some circumstances but not others? What signals register DNA damage and how do they impinge on the effector pathways of apoptosis? Is there a suborganellar apoptosome complex effecting the integration of death signals within the nucleus, just as there is in the cytoplasm? And what are the consequences of failure to initiate apoptosis in response to DNA damage? PMID- 11048729 TI - Corpse clearance defines the meaning of cell death. AB - While philosophers seek the meaning of life, cell biologists are becoming ever more interested in the meaning of death. Apoptosis marks unwanted cells with 'eat me' signals that direct recognition, engulfment and degradation by phagocytes. Far from being the end of the story, these clearance events allow scavenger cells to confer meaning upon cell death. But if the phagocytic 'spin doctors' receive or transmit the wrong messages, trouble ensues. PMID- 11048730 TI - CD95's deadly mission in the immune system. AB - Apoptosis in the immune system is a fundamental process regulating lymphocyte maturation, receptor repertoire selection and homeostasis. Thus, death by apoptosis is as essential for the function of lymphocytes as growth and differentiation. This article focuses on death receptor-associated apoptosis and the role of CD95 (Apo-1/Fas)-mediated signalling in T-cell and B-cell development and during the course of an immune response. Gaining an insight into these processes improves our understanding of the pathogenesis of diseases such as cancer, autoimmunity and AIDS, and opens new approaches to rational treatment strategies. PMID- 11048731 TI - Apoptosis in development. AB - Essential to the construction, maintenance and repair of tissues is the ability to induce suicide of supernumerary, misplaced or damaged cells with high specificity and efficiency. Study of three principal organisms--the nematode, fruitfly and mouse--indicate that cell suicide is implemented through the activation of an evolutionarily conserved molecular programme intrinsic to all metazoan cells. Dysfunctions in the regulation or execution of cell suicide are implicated in a wide range of developmental abnormalities and diseases. PMID- 11048732 TI - Apoptosis in the nervous system. AB - Neuronal apoptosis sculpts the developing brain and has a potentially important role in neurodegenerative diseases. The principal molecular components of the apoptosis programme in neurons include Apaf-1 (apoptotic protease-activating factor 1) and proteins of the Bcl-2 and caspase families. Neurotrophins regulate neuronal apoptosis through the action of critical protein kinase cascades, such as the phosphoinositide 3-kinase/Akt and mitogen-activated protein kinase pathways. Similar cell-death-signalling pathways might be activated in neurodegenerative diseases by abnormal protein structures, such as amyloid fibrils in Alzheimer's disease. Elucidation of the cell death machinery in neurons promises to provide multiple points of therapeutic intervention in neurodegenerative diseases. PMID- 11048733 TI - From bench to clinic with apoptosis-based therapeutic agents. AB - A retrospective look at the basis of human disease pathogenesis almost always reveals an apoptotic component that either contributes to disease progression or accounts for it. What makes this field particularly exciting is the breadth of therapeutic opportunities that are on offer. The pace of apoptosis research has raised expectations that therapeutics will follow soon. But many of the organizations that are best placed to take advantage of these discoveries consider the ability to modulate the life or death of a cell for the purpose of disease treatment as perhaps being 'too good to be true'. Nevertheless, practical therapeutics that modulate apoptosis will no doubt appear in the clinic or on the shelf in the next few years. PMID- 11048734 TI - Analyses of butyrophenones and their analogues in whole blood by high-performance liquid chromatography-electrospray tandem mass spectrometry. AB - Five butyrophenones and two analogues contained in human whole blood have been analyzed by high-performance liquid chromatography (HPLC)-electrospray (ES) tandem mass spectrometry (MS). All compounds gave the base peaks due to [M+1]+ by HPLC-ES-single MS. The product ions formed from each quasi-molecular ion by HPLC ES-tandem MS showed the base peaks at m/z 165 for four compounds. The mass chromatography of HPLC-ES-tandem MS showed much higher sensitivity than that of HPLC-ES-single MS for all drugs spiked to whole blood. Therefore, regression equations, detection limits, recovery rates and precision were studied for haloperidol, bromperidol and fluoropipamide spiked to human whole blood by means of mass chromatography of HPLC-ES-tandem MS. The three compounds showed good linearity in the range of 0.2-0.8 ng/ml with a detection limit of about 0.1 ng/ml. Recoveries of the three compounds spiked to whole blood (0.2 and 0.8 ng added to 1 ml whole blood) were 23.6-81.2%; the coefficients of intra- and inter day variations were 8.4-10.4 and 14.5-17.5%, respectively. The three compounds in whole blood could be actually determined 3 and 6 h after oral administration of 1 mg each of haloperidol and bromperidol, and 10 mg of floropipamide in a volunteer. PMID- 11048735 TI - Smoking and oxidant stress: assay of isoprostane in human urine by gas chromatography-mass spectrometry. AB - Isoprostane (8-epi-prostaglandin F2alpha) is synthesized non-enzymatically from arachidonate and active oxygen. We examined the relationship of smoking and excretion of isoprostane in urine with gas chromatography-mass spectrometry selected ion monitoring assay and the stable isotope dilution method. Urine isoprostane concentrations were significantly higher in smokers (n=81, 605.24+/ 59.01 ng/mg creatinine) than in non-smokers (n=39, 424.07+/-70.37 ng/mg creatinine), but concentrations in ex-smokers (n=21, 487.27+/-98.48 ng/mg creatinine) did not differ significantly from those in the other groups. In smokers, age, the duration of smoking, and the number of cigarettes per day were not correlated with urine isoprostane concentrations. However, urine isoprostane concentrations were negatively correlated with time since quitting in ex-smokers and with age in non-smokers. These results indicate that smoking increases isoprostane concentration in urine and suggest that smoking causes lipid peroxidation by oxidant stress. PMID- 11048736 TI - Rapid and sensitive quantification of 8-isoprostaglandin F2alpha in human plasma and urine by liquid chromatography-electrospray ionization mass spectrometry. AB - The isoprostane, 8-isoprostaglandin F2alpha (8-iso-PGF2alpha), is produced non enzymatically by direct oxidation of arachidonic acid on the cell surface by oxygen radicals. We developed a new assay method for 8-iso-PGF2alpha using 2H4-8 iso-PGF2alpha as the internal standard (I.S.) by high-performance liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS). For this assay, we established a very simple and rapid pretreatment method using a membrane filter-type solid-phase extraction column (Empore disk cartridge) for human urine extracts or intact plasma. LC-ESI-MS was performed in the selected ion monitoring (SIM) mode using target ions at m/z 353.24 (8-iso-PGF2alpha) and m/z 357.26 (I.S.) with a resolution of 1,500. The imprecision for this method was below 13.7%. Mean inaccuracy was 8.7% for added levels of 8-iso-PGF2alpha up to 5,000 pg/ml of urine and 500 pg/ml of plasma. Determination of plasma and urinary 8-iso-PGF2alpha concentrations in healthy subjects by the present method revealed that its urinary concentration in smokers tends to be higher than that in nonsmokers. PMID- 11048737 TI - Metabolism of an ionic contrast medium and the related agents. AB - Liquid chromatography-atmospheric pressure chemical ionization mass spectrometry was applied to analyze the iodinated compounds and their glycine conjugates. The negative-ion mass spectra of the iodinated compounds gave [M-H]-, [M-COOH]- and [I]- ions. The positive- and negative-ion mass spectra of the glycine conjugates showed abundant [M+H]+ and [M-H]- ions. Fragmentations of the glycine conjugates obtained in the positive-ion mode were different from those in the negative-ion mode, the former providing more useful structural information for the presence of glycine. Mouse kidney mitochondria were more active in glycine conjugation than liver mitochondria. Mono-substituted benzoic acids were conjugated with glycine in liver and kidney, whereas the acids having three functional groups or more did not undergo glycine conjugation in liver and kidney. PMID- 11048738 TI - Analysis of citrulline in rat brain tissue after perfusion with haloperidol by liquid chromatography-mass spectrometry. AB - We have investigated the potential of high-performance liquid chromatography (HPLC) coupled to mass spectrometry (MS) to determine enrichments of citrulline as a marker for in vivo nitric oxide (NO) production in brain tissue. The analysis of citrulline as the butyl ester derivative was evaluated using two types of ionization: electron spray ionization (ESI) and atmospheric pressure chemical ionization (APCI). APCI-MS appeared to be more suitable for determination of citrulline than ESI-MS, because the ion intensity of the protonated molecule ion [M+H]+, m/z 232, of citrulline in the former was about twelve times higher than in the latter. The chromatography was carried out on a reversed C8 column with the mobile phase consisting of 15% acetonitrile: 85% H2O: 0.2% acetic acid (v/v). The calibration curve had good linearity within the concentration range investigated (5 ng to 500 ng/ml). The limit of determination was estimated to be ca. 1 ng/ml of standard solution. The method was applied to the analysis of citrulline in the brain dialysate obtained from rat after perfusion of the striatum with haloperidol (HP, 0.1 mM). It is concluded that APCI-MS in combination with HPLC can be successfully applied to determination of citrulline in brain tissue, thus providing a useful tool for assessment of in vivo NO production. PMID- 11048739 TI - Inborn errors of metabolism discovered in Asian department of pediatrics and mental retardation research center. AB - To heighten the effectiveness of chemical diagnosis for inborn errors of metabolism (IEM) using urease pretreatment and GC-MS analysis, a sample collection and transportation method was contrived. The resulting "filter paper set" allows simple urine collection and transportation, and enables anyone from anywhere to receive the GC-MS analysis without the limitations of place or time. Using filter paper sets, high-risk screening of undiagnosed children or mentally retarded children with unknown cause was conducted in cooperation with hospitals and universities in several Asian countries. During 8 months 203 patients from China and India were analyzed and 20 cases of IEM were chemically diagnosed. These diagnoses greatly contributed to the treatment of children with intractable diseases who lived in Asian countries where analytical techniques and facilities for IEM were not sufficient. PMID- 11048740 TI - Detection and characterization of modified nucleosides in serum and urine of uremic patients using capillary liquid chromatography-frit-fast atom bombardment mass spectrometry. AB - To determine RNA metabolism in uremia, capillary liquid chromatography-frit-fast atom bombardment mass spectrometry was employed for the characterization of ribonucleosides in serum and urine of uremic patients, and the profiles were compared with those of healthy subjects. We have characterized 20 nucleosides in serum and 23 nucleosides in urine from both healthy subjects and uremic patients; most of them were modified nucleosides derived from tRNA breakdown products. Four metabolites derived from allopurinol were detected as exogenous nucleosides in patients receiving allopurinol; these include allopurinol-1-riboside, oxipurinol 1-riboside, oxipurinol-7-riboside and a unknown oxipurinol riboside. The endogenous and exogenous ribonucleosides were retained at higher levels in uremic serum, and may play a contributory role as toxins responsible for clinical symptoms of uremia. PMID- 11048741 TI - Gas chromatography-mass spectrometry with tert.-butyldimethylsilyl derivation: use of the simplified sample preparations and the automated data system to screen for organic acidemias. AB - A simplified, sensitive screening method for organic acidemias by gas chromatography-mass spectrometry using urease/direct preparations and tert. butyldimethylsilylation (TBDMS) instead of trimethylsilylation (TMS) is described. We compiled GC-MS data on TBDMS derivatives, including methylene unit values, and quantifying and confirming ions, for use in the automated data system we developed. Quantification using [M-57]+ ions by mass chromatography was more sensitive, and the coefficient variation was smaller, compared with TMS derivatives. We confirmed the usefulness of this system, analyzing urine specimens from 53 patients with 15 different disorders. PMID- 11048742 TI - Rapid, simplified and sensitive method for screening fructose-1,6-diphosphatase deficiency by analyzing urinary metabolites in urease/direct preparations and gas chromatography-mass spectrometry in the selected-ion monitoring mode. AB - Children with fructose-1,6-diphosphatase (FDPase) deficiency often experience life threatening episodes such as ketotic hypoglycemia. We report here a rapid, simplified and sensitive method to analyze glycerol-3-phosphate (G3P) and glycerol in urine, that can be used to detect FDPase deficiency. We used the urease/direct preparation and gas chromatography-mass spectrometry in the selected-ion monitoring mode, enabling detection of G3P and glycerol level in normal controls. Using this approach, FDPase deficiency can be more easily diagnosed and differentiated from glycerol kinase deficiency or glycerol infusion patients. To date, diagnosis has been essentially based on the assay of enzymes in the liver. The proposed non-invasive method provides a clinically significant diagnostic tool that may help prevent episodic attacks. PMID- 11048743 TI - Determination of ionization efficiency of glycated and non-glycated peptides from the N-terminal of hemoglobin beta-chain by electrospray ionization mass spectrometry. AB - We compared the ionization efficiency of glycated and non-glycated peptides for the HbA1c measurement method developed by Kobold et al. [Clin. Chem., 43 (1997) 1944] based on LC-ESI-MS analysis of the N-terminal peptides of the beta-chains released by cleavage of the hemoglobin with endoproteinase Glu-C. Taking half the peak area of the doubly charged ion and adding it to the area of the singly charged ion, we determined that the slope of the resulting calibration curve was nearly equal to 1, and the reproducibility of the added values was better than the values calculated by the doubly or the singly charged ion alone. PMID- 11048744 TI - Analysis of glutathionyl hemoglobin levels in diabetic patients by electrospray ionization liquid chromatography-mass spectrometry: effect of vitamin E administration. AB - By using electrospray ionization liquid chromatography-mass spectrometry, we demonstrated that glutathionyl hemoglobin (Hb)beta-chain levels are markedly increased in the erythrocytes of diabetic patients as compared with healthy subjects. The administration of vitamin E to the diabetic patients for 8 weeks significantly decreased the levels of glutathionyl Hbbeta, whereas it did not affect the levels of HbA1c, glycated Hbbeta or glycated Hbalpha. Glutathionyl Hb levels can be used as a new clinical marker of oxidative stress. PMID- 11048745 TI - Specific determination of urinary 6beta-hydroxycortisol and cortisol by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry. AB - The urinary 6beta-hydroxycortisol to cortisol ratio is believed to be a noninvasive index of cytochrome P450 3A activity. For precise assessment of the ratio in human urine, we have developed a reversed-phase high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry method. The selective method was accurate and reproducible with intra- and inter-day precision of variation coefficients of less than 8%. The 6beta-hydroxycortisol to cortisol ratio ranged from 3.0 to 12.4 in healthy Japanese 24-h urine. With the recent popularization of LC-MS, our LC-MS method will be advantageous to detect human in vivo CYP3A activity for clinical investigation and routine measurement in various laboratories. PMID- 11048746 TI - Morphometric study of nucleus ambiguus in multiple system atrophy presenting with vocal cord abductor paralysis. AB - AIM: To identify lesions responsible for vocal cord abductor paralysis (VCAP) in multiple system atrophy (MSA), we performed a morphometric study of the nucleus ambiguus which innervates the intrinsic laryngeal muscles. METHODS: Two autopsied cases of MSA presenting with VCAP and one control were examined. Both cases of MSA showed selective neurogenic atrophy of the posterior cricoarytenoid muscles among the intrinsic laryngeal muscles, while no abnormalities were seen in the control. From a block of the medulla oblongata, sections 10 microm thickness were cut serially without spacing and stained with cresyl violet. The ambiguus neurons were counted in all the sections to make a histogram. RESULTS: In the control case, ambiguus neurons showed densely populated areas and sparsely populated areas alternately with significant difference in the mean neuronal density between two areas. In MSA, ambiguus neurons were significantly decreased in number at all levels. It indicates that the neurogenic atrophy of the posterior cricoarytenoid muscle is derived from the neuronal loss of the nucleus ambiguus. CONCLUSION: Though it has still been controversial whether or not the ambiguus neurons are decreased in number in MSA with VCAP, we speculated possible reasons for the disagreement on the involvement of the nucleus ambiguus as follows: different mechanism of VCAP are playing role, and histometric data have been disturbed by factors such as split-cell counting error and marked variation in the distribution of the ambiguus neurons. PMID- 11048747 TI - Infantile neuroaxonal dystrophy and giant axonal neuropathy--overlap diseases of neuronal cytoskeletal elements in childhood? AB - Giant axonal neuropathy (GAN) and infantile neuroaxonal dystrophy (INAD) are two progressive neurodegenerative disorders of childhood that have considerable clinical as well as histological overlap but are believed to be ultrastructurally distinct. The clinicopathological and ultrastructural features of three cases of INAD, two of whom are siblings and one case of GAN are described. The sural nerve biopsies in all four cases were essentially similar on light microscopy revealing giant axons. On electron microscopy, the findings in the case of GAN were typical with dense accumulation of neurofilaments within the giant axons. In the three cases of INAD, too, in addition to accumulation of mitochondria and organelles with vesiculotubular profiles, a similar increase in neurofilaments was evident. We, therefore, believe that these two disorders may represent a spectrum in evolution of intermediate filament pathology with various organelles participating in the temporal evolution of the disease process. PMID- 11048749 TI - A destructive hemispheric cleft discovered in a female fetus. AB - The case of a female fetus who died at the 38th week of gestation is reported. Neuropathological examination disclosed a hemispheric cleft located in the medial part of the left frontal lobe. The cleft did not extend to the ventricle and was not a true schizencephaly. Furthermore, a focus of gliosis was evidenced by GFAP staining located in the superior bank of the cleft suggesting a destructive origin of the process. Maternal psychological stress occurring during the middle of the pregnancy could have favored hemodynamic disturbances acting on the fetal circulation. PMID- 11048748 TI - Correlation of tumor p53 and PCNA with response and survival of glioblastoma in patients treated with an ECOG protocol of pre-irradiation chemotherapy. AB - BACKGROUND: The ability to predict treatment responsiveness and survival of patients with glioblastoma multiforme, the most malignant and most common primary brain tumor, would be a valuable asset. Tumor and proliferation markers such as p53 and PCNA have been immunohistochemically defined and have been useful in other tumors in determining prognosis. Therefore, the authors studied the correlation of responsiveness to treatment, time to progression and survival with p53 and PCNA labeling indices in a pre-irradiation chemotherapy study of the glioblastoma multiforme. METHODS: Immunohistopathology for labeling indices for p53 and PCNA using formalin-fixed, paraffin-embedded tissue from the glioblastomas of 23 patients entered into a phase II ECOG trial of pre irradiation chemotherapy were defined using the streptavidin-peroxidase technique with AEC chromogen. The labeling indices were correlated with response to treatment time to progression and overall survival. Most patients received three cycles of BCNU for three days over three months and cisplatin monthly for three days over three months prior to external beam irradiation. RESULTS: There were no significant differences in treatment response, time to progression or overall survival in glioblastoma, patients with positive p53 labeling index (> 5%) versus a negative p53 labeling index (< or = 5%) or positive PCNA labeling (> 10%) versus a negative labeling index (< or = 10%) or any combination of P53 and PCNA labeling indices. CONCLUSIONS: Using this protocol of pre-irradiation chemotherapy, p53 and PCNA labeling indices in the glioblastoma multiforme did not predict treatment benefit. PMID- 11048752 TI - History of the Department of Neuropathology in Hamburg. PMID- 11048750 TI - Pleomorphic xanthoastrocytoma of the cerebellum. AB - We report a clinicopathologic case of a pure cerebellar pleomorphic xanthoastrocytoma occurring in a 68-year-old male patient. The occurrence of pleomorphic xanthoastrocytoma outside the cerebral hemispheres is exceedingly rare. In the cerebellum only five cases have been reported so far, four of which are composite pleomorphic xanthoastrocytoma-gangliogliomas. This observation reinforces the argument that pleomorphic xanthoastrocytomas should be included in the differential diagnosis of cerebellar neoplasms. PMID- 11048753 TI - The role of microglia cells in brain diseases. PMID- 11048754 TI - Clinical and laboratory diagnosis of Creutzfeldt-Jakob disease. PMID- 11048755 TI - Pathology of Jakob-Creutzfeldt disease. PMID- 11048756 TI - Psychiatric aspects of alcoholism. PMID- 11048757 TI - Neuropathology of chronic alcoholism. PMID- 11048758 TI - Subcortical angiopathic encephalopathy: cerebral small vessel diseases and differential diagnosis on MRI. PMID- 11048759 TI - Neuropathology of hereditary subcortical angiopathic encephalopathy. PMID- 11048760 TI - Surgical therapy of gliomas. PMID- 11048761 TI - Immunobiology of neuroepithelial tumors. PMID- 11048762 TI - Pre-natal sensitization in humans. AB - Allergy has a very strong hereditary component but even in identical twins, concordance for the development of allergic disease can be as low as 50%. This suggests that there is a very strong environmental influence on manifestations of sensitization. To what extent environment might have an influence on the ontogeny of sensitization antenatally has hitherto not been a focus of much research. However, circumstantial evidence suggests that this may be important. PMID- 11048763 TI - Primary sensitization to inhalant allergens. AB - The neonatal T-cell system is capable of responding to allergens at birth, indicating the occurrence of prenatal sensitization, and the cytokine profile of these responses is skewed towards the Th-2 type. This response is further modified by postnatal exposure to different types of allergens. In relation to inhalant allergen (employed by HDM) the low level fetal Th-2 responses in non atopics appear to be down-regulated rapidly after birth, parallel to an increase in allergen-specific IFN-gamma production. In contrast, atopics appear to consolidate their initial Th-2 responses, and around the age of 6 exhibit a cytokine response profile similar to the adult pattern. A pre-existing deficiency in IFN-gamma production may be one of the key factors determining the postnatal persistence of Th-2 responses in atopics. PMID- 11048764 TI - Progression from allergic sensitization to asthma. AB - Asthma is one of the atopic diseases strongly associated with allergy. High aeroallergen exposure in the immediate postnatal period has been associated with higher risk of sensitization and chronic asthma. It is proposed that following in utero allergen sensitization, postnatal high dose allergen exposure localizes inflammation to the airways. In association with adjuvantizing effects of some virus infections, eosinophils and neutrophils are recruited which contribute to epithelial damage and the initiation of the remodelling process. Eventually, the latter processes lead to sufficient airway narrowing to manifest as the first symptoms of asthma. Thus, the immunopathology of asthma is fully established by the time of first symptoms and future strategies will need to identify those at risk of developing the disease before irreversible changes in the airways are established. PMID- 11048765 TI - Infant lung function, bronchial responsiveness and the development of asthma. AB - Infants who wheeze are likely to have narrowed or overly compliant airways rather than atopy. After around 3 years of age, airway responsiveness and atopy are associated with the development of wheeze and the diagnosis of asthma. Anti- or proinflammatory genes are likely to be responsible for at least part of the predisposition to wheeze seen in older children, but further studies are needed to clarify this situation. PMID- 11048766 TI - Allergic diseases in farmers' children. AB - Several studies have reported lower rates of allergic sensitization and allergies in children living in rural as compared to urban communities. This has been attributed to the lower levels of air pollution in rural areas. The question arises whether other factors in the rural environment could explain the lower prevalence rates of allergic sensitization and hay fever. A first report from rural South Bavaria in Germany demonstrated that children living in a home where coal and wood were used for heating had a significantly lower risk of suffering from hay fever (odds ratio 0.57 (0.34-0.98)), of being sensitized to common allergens (OR 0.67 (0.49-0.93)) and of having bronchial hyperresponsiveness (OR 0.55 (0.34-0.90)) than their peers living in homes with other heating systems. Subsequently, the Swiss Study on Childhood Allergy and Respiratory Symptoms with Respect to Air Pollution (SCARPOL) tested the hypothesis that farming as parental occupation was associated with a lower risk of hay fever and atopy. A total of 1620 (86.0%) 6-15-year-old schoolchildren living in three rural communities of Switzerland were examined using a standardized questionnaire completed by the parents and IgE antibodies against six common aeroallergens in serum samples of 404 (69.3.0%) of the 13-15-year-olds. Farming as parental occupation was significantly associated with lower rates of reported hay fever symptoms and allergic sensitization. Comparing children from farming with those from non farming environments, the adjusted OR was 0.34 (95% CI: 0.12-0.89) for sneezing attacks during the pollen season, and 0.31 (95% CI: 0.13-0.73) for a sensitization to allergens. These results have recently been confirmed in a new and much larger survey in rural South Bavaria. Several alternative explanations have to be considered when interpreting these findings, namely, selection bias, the development of tolerance, increased microbial stimulation and a more traditional lifestyle (diet and housing conditions). Based on present knowledge, the underlying environmental factor explaining the protective effect of the farming environment has not yet been identified. PMID- 11048767 TI - The role of ozone. PMID- 11048768 TI - Can immunization affect the development of allergy? AB - A changing pattern of infections may be of importance for the increase in prevalence of asthma and other allergic diseases in developed countries during recent decades. The clear inverse relation between number of siblings and atopy observed in several studies may be related to a protective role of infections, although specific information is inconclusive. A recent study showed that positive tuberculin responses in schoolchildren correlated with a lower prevalence of atopic disorders, but other studies did not find a relation between BCG vaccination and allergic disease or sensitization. Transient production of IgE antibodies to pertussis toxin has been demonstrated after pertussis immunization; however, randomized clinical trials involving both whole cell and acellular pertussis vaccines have failed to show any enhancement of atopic manifestations in children. Epidemiologic investigations indicate that viral infections may either promote (RSV) or inhibit (hepatitis A, measles) atopy, although data are scarce. In conclusion, the evidence is limited regarding a direct role of vaccinations for development of atopic manifestations, but speaks against a major effect of some types of vaccinations. On the other hand, since some infections may offer protection in relation to allergy, vaccination could result in an increased risk. PMID- 11048769 TI - Fatty acids and atopic disease. PMID- 11048770 TI - Diet, infection and wheezy illness: lessons from adults. AB - An increase in asthma and atopic disease has been recorded in many countries where society has become more prosperous. We have investigated two possible explanations: a reduction in childhood infections and a change in diet. In a cohort of people followed up since 1964, originally selected as a random sample of primary school children, we have investigated the relevance of family size and the common childhood infectious diseases to development of eczema, hay fever and asthma. Although membership of a large family reduced risks of hay fever and eczema (but not asthma), this was not explained by the infections the child had suffered. Indeed, the more infections the child had had, the greater the likelihood of asthma, although measles gave a modest measure of protection. We have investigated dietary factors in two separate studies. In the first, we have shown the risks of bronchial hyper-reactivity are increased seven-fold among those with the lowest intake of vitamin C, while the lowest intake of saturated fats gave a 10-fold protection. In the second, we have shown that the risk of adult-onset wheezy illness is increased five-fold by the lowest intake of vitamin E and doubled by the lowest intake of vitamin C. These results were supported by direct measurements of the vitamins and triglycerides in plasma. We have proposed that changes in the diet of pregnant women may have reflected those observed in the population as a whole and that these may have resulted in the birth of cohorts of children predisposed to atopy and asthma. The direct test of this is to study the diet and nutritional status of a large cohort of pregnant women and to follow their offspring forward. This is our current research. PMID- 11048771 TI - What are the candidate groups for pharmacotherapeutic intervention to prevent asthma? AB - Despite the development of many new therapies for the treatment of asthma, the prevalence of this disease is still increasing in many areas of the world. Today no intervention is able to completely cure asthma but chronic therapies could decrease its severity. Moreover, asthma is one of the most common chronic diseases of childhood and its pharmacoeconomic burden is huge. Epidemiologic studies have identified some of the major factors involved in the pathogenesis and evolution of asthma. Several prevention programs have been developed in different countries with various success rates. Most of those interventions were based on allergen avoidance. From studies aimed at controlling early asthma and from epidemiologic data, we have learned to identify high-risk groups, e.g, the atopic child with allergic asthma, with a family history of asthma or allergy related disease and early sensitization to aeroallergens. Only a few prospective studies aimed at preventing the onset of asthma have been published. With ketotifen, Iikura et al. could prevent the onset of asthma after a 1-year period in patients suffering from atopic dermatitis. Another study has been published by Bustos et al. concerning children with a family history of allergy and high total IgE levels. Those studies involved about 100 patients. No follow-up data has been published for either of them. Recently, the first results from the ETAC (Early Treatment of the Atopic Child) trial have been reported. This study involved 817 atopic children with atopic dermatitis and a family history of atopy: cetirizine halved the number of patients developing asthma in the subgroups (200 children) sensitized to house dust mite (51.5% versus 28.6%) or pollen (58.8% versus 27.8%). The optimal target for pharmaceutical intervention to prevent asthma would seem to be high risk patients: children with atopic dermatitis, a family history of asthma or atopic disease and early sensitization to aeroallergens. Primary prevention in whole populations (e.g. starting even before the onset of atopic dermatitis or allergen sensitization) does not at present appear to be a realistic approach. PMID- 11048772 TI - Computer applications in the interpretation of the exercise electrocardiogram. AB - The exercise electrocardiogram remains the noninvasive diagnostic test of first choice in patients with coronary artery disease. While new technology offers novel diagnostic possibilities and the ability to assess patients unsuitable for exercise testing, no other investigation has to this point furnished the quality of functional information and value-for-predictive accuracy of exercise electrocardiography. In this article, we describe how this central position in the work up of the cardiac patient has been secured through the evolution of the microprocessor. Particularly important has been its ability to harness and present large volumes of raw data, to derive and manipulate multivariate equations for diagnostic prediction, and to run 'expert' systems which can pool demographic and exercise test data, calculate risk scores, and prompt the nonexpert with advice on current management. These key features explain the pivotal role of the exercise test in the diagnostic, and increasingly prognostic, armoury of the cardiovascular clinician. PMID- 11048774 TI - Therapeutic impact of exercise on psychiatric diseases: guidelines for exercise testing and prescription. AB - Aerobic exercise seems to be effective in improving general mood and symptoms of depression and anxiety in healthy individuals and psychiatric patients. This effect is not limited to aerobic forms of exercise. There are almost no contraindications for psychiatric patients to participate in exercise programmes, provided they are free from cardiovascular and acute infectious diseases. However, very little is known about the effects of exercise in psychiatric disease other than those in depression and anxiety disorders. A few reports indicate the need for controlled investigations in psychotic and personality disorders. Unfortunately, no general concept for a therapeutic application of physical activity has been developed so far. Reliance on submaximal measures is highly recommended for fitness assessment. Monitoring of exercise intensity during training sessions is most easily done by measuring the heart rate using portable devices (whereas controlling the exact workload may be preferable for scientific purposes). Appropriate pre- and post-training testing is emphasised to enable adequate determinations of fitness gains and to eventually allow positive feedback to be given to patients in clinical settings. PMID- 11048773 TI - Strength training in the elderly: effects on risk factors for age-related diseases. AB - Strength training (ST) is considered a promising intervention for reversing the loss of muscle function and the deterioration of muscle structure that is associated with advanced age. This reversal is thought to result in improvements in functional abilities and health status in the elderly by increasing muscle mass, strength and power and by increasing bone mineral density (BMD). In the past couple of decades, many studies have examined the effects of ST on risk factors for age-related diseases or disabilities. Collectively, these studies indicate that ST in the elderly: (i) is an effective intervention against sarcopenia because it produces substantial increases in the strength, mass, power and quality of skeletal muscle; (ii) can increase endurance performance; (iii) normalises blood pressure in those with high normal values; (iv) reduces insulin resistance; (v) decreases both total and intra-abdominal fat; (vi) increases resting metabolic rate in older men; (vii) prevents the loss of BMD with age; (viii) reduces risk factors for falls; and (ix) may reduce pain and improve function in those with osteoarthritis in the knee region. However, contrary to popular belief, ST does not increase maximal oxygen uptake beyond normal variations, improve lipoprotein or lipid profiles, or improve flexibility in the elderly. PMID- 11048776 TI - Commotio cordis: an underappreciated cause of sudden death in athletes. AB - Over the last few years, the recognised cardiovascular risks of sporting activities have been extended to include cardiac arrest resulting from low-energy precordial chest impact produced by projectiles (e.g. baseball) or bodily contact, in the young, healthy and active athlete [also known as commotio cordis (CC)]. However, case reports of CC in European medical literature can be traced back for at least 130 years. CC accounts for a small, but important, subset of sudden death during sporting activities. It is a devastating electrophysiological event in the young athlete, and one which has generated considerable concern, both in the medical profession as well as in the public. The mechanism of sudden death appears to be caused by ventricular fibrillation, which occurs when the chest impact is delivered within a narrow, electrically vulnerable portion of the cardiac cycle, that is, during repolarisation, just before the peak of the T wave. Resuscitation of these victims is possible with prompt cardiopulmonary resuscitation and defibrillation. Preventive measures, such as the use of age appropriate safety baseballs and suitably designed chest wall protection, may reduce the risk of sudden death and, thus, make the athletic field a safer place for young athletes. PMID- 11048777 TI - Separation and identification of volatile components in the fermentation broth of Trichoderma atroviride by solid-phase extraction and gas chromatography-mass spectrometry. AB - A preseparated fermentation broth of Trichoderma atroviride strain 11 is analyzed by gas chromatography followed by mass-spectral detection using a Finnigan MAT GCQ apparatus. After preseparation in a C18 and a silica gel column, nineteen pyrone and dioxolane derivatives and two aliphatic esters are obtained, respectively. Among these, the four dioxolane derivatives have not been identified previously. The main component is found to be 5,5'-dimethyl-2H-pyran-2 on. The relative standard deviation for the determination of the retention time and the peak area (measured in ion counts) is 0.1% and 4.5%, respectively. PMID- 11048775 TI - The physiology of the highly trained female endurance runner. AB - Continuing improvements in the performance of female endurance runners and increasing levels of participation have generated the need to know more about the physiology of this group. Specific research is needed in this area, as data referring to male endurance runners cannot legitimately be applied to the female endurance runner because of their markedly different physiological and hormonal profiles. Recent developments in our understanding of an athlete's physiology (mainly in relation to the male endurance runner) have revealed new areas of interest that need to be assessed with specific reference to the female athlete. Relatively little attention has been directed towards identifying the major physiological characteristics of the highly trained/elite female endurance runner in general, and that which has been published on such factors and the effects of the menstrual cycle have produced equivocal results. Moreover, the impact of such training upon the menstrual cycle and endurance running performance is a controversial area, especially when assessing its subsequent impact on health related issues. Reports of the condition referred to as the 'female athlete triad' have increased in recent years, with a decrease in bone mineral density predisposing the female athlete to increased risks of stress fractures. The aetiology of this triad is multifactorial, with such risk factors including nutrition, menstrual status, training intensity and frequency, body size and composition and psychology/physical stress. However, research limitations and flaws have lead to controversy in the literature regarding the immediate and long term effects of the triad on the female athlete. Likewise, the effects of the oral contraceptive pill on health and endurance performance also remain elusive, with a dearth of research pertaining to how oral contraceptive agents can aid athletic performance and the long term health of the female athlete. The purpose of this paper is to critically appraise the existing literature to provide a current review of the physiological scientific knowledge base in relation to the female athlete, health, training and performance, with suggestions for future areas of research. It is well known that certain menstrual and health-related performance factors of the female athlete, that is, physiological predictors of performance and body fat, have been extensively investigated over the last 30 years. However, a variety of methodological flaws and inconsistencies are present within the research and thus only the most prominent and well controlled studies within this area over the past 30 years will be referred to. PMID- 11048778 TI - Deconvolution of composite chromatographic peaks by simultaneous dual detections. AB - Composite chromatographic peaks are deconvoluted by a method that uses ratio formation from signals of simultaneous double detection. The method is generally suitable if two detector signals can simultaneously be acquired and their uses do not need any a priori assumption or mathematical shape analysis. A simple deduction makes the compound- and detector-specific intensive parameters explicit, which allows for the digital construction of directly invisible component peaks. The simultaneous double detection is shown to be superior to coupled detectors, sequentially fixed chromatograms, and subsequently synchronized peaks. The combination of circular dichroism and ultraviolet (UV) detection is shown to be especially advantageous in the analysis of enantiomers for which the other ratio-forming peak-deconvolution techniques have inherently been insensitive. The double chiroptical UV detection can be of further use to decompose overlapping peaks of nonenantiomeric compounds that are highly similar. The capacity of the method is exemplified by the analysis of morphine alkaloids, steroid oximes, and synthetic heterocycles. PMID- 11048779 TI - Optical resolution of a series of potential cholecystokinin antagonist 4(3H) quinazolone derivatives by chiral liquid chromatography on alpha1-acid glycoprotein stationary phase. AB - Optical resolution of the enantiomers of new 4(3H)-quinazolone derivatives is investigated using the alpha1-acid glycoprotein chiral stationary phase (Chiral AGP). Stereoselective separation of the model compounds can be controlled by varying the pH and adding uncharged organic modifiers (acetonitrile and 2 propanol) to the mobile phase. For the majority of quinazolone derivatives, Chiral-AGP is proved to be an excellent enantioselector, because optimized chromatographic conditions allow for the baseline separation of the enantiomers. Separation factors between 1.19 and 1.85 are obtained. The effects of acetonitrile and 2-propanol on the chromatographic behavior of the model compounds are quite different because of their different hydrophobic- and hydrogen-bonding properties. The eluent pH and organic modifier concentration also contributes to the chiral recognition by altering the protein environment. The analysis of the experimental results leads to new information about the chromatographic mechanism on a Chiral-AGP surface. PMID- 11048780 TI - Enantioselective assay of chloroquine and its main metabolite deethyl chloroquine in human plasma by capillary electrophoresis. AB - A sensitive assay for the determination of chloroquine (Clq) and its pharmacologically active metabolite deethyl chloroquine in plasma by capillary electrophoresis (CE) is developed. Plasma levels of drug and metabolite are measured using HeCd laser-induced fluorescence (LIF) detection over a range of three orders of magnitude from 2 to 1000 ng/mL after liquid-liquid extraction. A limit of detection of 0.5 ng/mL is achieved. Validation of the method yields intra- and interday precision data within the limits of 10% (20% at limit of quantitation) and intra- and interday accuracy data greater than 6% throughout the whole working range. The method is applied for the drug monitoring of patients treated with Clq. Based upon this assay, two enantioselective CE-LIF methods for Clq and its main metabolite are developed. Mixtures of substituted gamma-cyclodextrins are used as chiral selectors. A baseline separation of the enantiomers of both analytes in one run is achieved in less than 11 min (method A) and less than 9 min (method B), respectively. Hydroxychloroquine is used as the internal standard for both methods. PMID- 11048781 TI - Development and evaluation of a new method for the determination of the carotenoid content in selected vegetables by HPLC and HPLC-MS-MS. AB - Epidemologic studies have shown inverse correlation between the consumption of carotenoid-rich vegetables and the incidence of cancer. Therefore, analytical techniques for the quantitative determination of carotenoids in complex sample matrices are important. The most used method is reversed-phase (RP)-high performance liquid chromatography (HPLC). In this study, seventeen mobile-phase systems described in the literature and six RP-HPLC columns with differences in particle size and porosity are evaluated. Derived from these results, a new mobile-phase (acetonitrile, methanol, chloroform, and n-heptane) including solvent modifiers is presented, which allows an improved and more efficient separation of carotenoids. From all columns tested, the best chromatographic parameters are found using a silica C18 column (250 x 2 mm, 5 microm, 100 A). As was found, absorbance detection at 450 nm allows the determination of the carotenoids down to the picogram range with good linearity (R2 > 0.98). For the identification and quantitation of carotenoids in complex sample matrices (containing additionally other ultraviolet-absorbing compounds), the optimized RP chromatographic system is coupled to a mass spectrometer (MS) using an atmospheric pressure ionization interface. The calibration plots show high linearity (R2 > 0.99), and the detection limit is found in the lower nanogram range. Furthermore, collision-induced dissociation in the ion source allows for the identification of carotenoids by their characteristic fragmentation pathways. In this study, a total of nine species of vegetables commonly consumed in Central Europe are analyzed for their contents of carotenoids (namely lutein, zeaxanthin, beta-cryptoxanthin, and beta-carotene) by RP-HPLC and RP-HPLC-MS-MS. It is found that good sources for lutein are spinach, kale, and broccoli, and sources for beta-carotene are broccoli, spinach, kale, carrots, and tomatoes. This new method is an improvement for the identification and quantitation of carotenoids in complex biological tissues. PMID- 11048783 TI - Evaluation of solid-phase microextraction for the study of protein binding in human plasma samples. AB - Solid-phase microextraction (SPME) in combination with capillary gas chromatography and a nitrogen-phosphorous detector is used to study protein binding in human plasma samples. Local anesthetics of the amide-type (ropivacaine, bupivacaine, mepivacaine, prilocaine, and lidocaine) are used as model compounds in this evaluation. Carbowax/divinylbenzene (CW/DVB), polyacrylate, and polydimethylsiloxane fibers are tested. Sampling on CW/DVB fibers give the highest recovery in plasma samples compared with other fibers. Ultrafiltrate spiked with each of the substances is used for the construction of calibration curves. The protein binding is investigated at four different total concentrations from 0.5 to 15.0 microM. The degree of protein binding increases when the solute concentration decreases. Protein binding of the five solutes is investigated at four pH levels (6.4, 7.4, 8.4, and 9.4). It is found that protein binding increased with increasing pH. The influence of temperature variation (from 32 degrees C to 40 degrees C) on protein binding is also investigated. The protein binding decreases when the temperature increases. The methodology is validated and good correlation and precision are obtained. Back-calculated quality control samples give accuracy within 20% of theoretical values for all five substances. This study shows that SPME as a sample-preparation method gives the same protein binding for the studied local anesthetics as that achieved using earlier presented methods. PMID- 11048782 TI - Comparative evaluation of four detectors in the high-performance liquid chromatographic analysis of chiral nonaromatic alcohols. AB - A comparative evaluation of ultraviolet, polarimetric, refractive index, and evaporative light-scattering detection coupled with high-performance liquid chromatography has been developed for the separation and quantitation of the enantiomers of chiral nonaromatic alcohols, some of which are intermediates in the synthesis of chiral drugs. (R,S)-3-tert-butylamino-1,2-propanediol; (R,S) glycidol; and (R,S)-1-(4-morpholino)-2-octanol are selected as model compounds in order to compare the detection sensitivity and the linearity of the response with the four detectors. Separation of the enantiomers is performed using chiral stationary phases in normal-phase liquid chromatography. A one-day validation is achieved for (S)-3-tert-butylamino-1,2-propanediol with each detector, and limits of quantitation are determined for the three compounds. Advantages and limitations of the four detectors are discussed. PMID- 11048785 TI - FID spiking can originate from a number of sources. PMID- 11048784 TI - What is meant by the term methylene selectivity and how does it influence what I observe in carring out reverse-phase separations? PMID- 11048786 TI - Latitude-correlated genetic polymorphisms: selection or gene flow? AB - Latitude-correlated polymorphisms can be due to either selection-driven evolution or gene flow. To discriminate between them, we propose an approach that studies subpopulations springing from a single population that have lived for generations at different latitudes and have had a low genetic admixture. These requirements are fulfilled to a large extent by Ashkenazi and Sephardi Jews. The original population lived at a latitude of 35 degrees N, where the Sephardis still live. The Ashkenazis, however, moved to a latitude of 50 degrees N, starting about 10 centuries ago. The present study examines 3 latitude-correlated polymorphisms: PGP, PGM1, and AHSG. We found that PGP*2 and AHSG*2 alleles most likely underwent selection-driven evolution, but that PGM1*ts allele was not similarly affected. Since temperature might have been considered a reasonable selective factor, we also studied a population living at >800 m above sea level from Aosta Valley (Italy). PMID- 11048787 TI - Ethnicity and malaria affect surname distribution in consenza Province (Italy). AB - Three samples of surnames reported in the telephone directory of Cosenza province, Italy, are used, singly and together, to detect the presence of genetic barriers and to analyze the genetic relationship between the Italian and the Italo-Albanian communities living in this area. The genetic structure of the population seems characterized by the distinction between the northern and southern regions of the province. The Sibari plain, endemic with malaria until recently, probably constituted a genetic barrier. In the southern region of the province, the valley along the Crati river (also occupied by malarial fenlands), constituted a genetic barrier between the northern Sila upland and the western coast. Surname similarities between Italians and Italo-Albanians could be the result of gene flow and/or an initial choice of similar surnames; the second possibility accords with the persistence of the Albanian cultural identity and the level of endogamy in Albanian communities. In fact, the coefficient of relationship by isonomy (Ri), which is significantly higher in Italo-Albanians than in Italians, may be the result of genetic isolation and endogamy. PMID- 11048788 TI - Frequency distribution of mitochondrial DNA haplogroups in Corsica and Sardinia. AB - Mitochondrial DNA (mtDNA) polymorphisms were analyzed by polymerase chain reaction amplification and haplogroup-specific restriction screening in populations from Corsica and Sardinia. These included 56 individuals from the area of Corte, central Corsica (France), 51 individuals from Gallura, northern Sardinia (Italy), and 45 individuals from Barbagia, central Sardinia. The screening revealed that about 95% of mtDNAs could be grouped in 8 of the 9 European haplogroups, including H-K, T-V, and X. Our results confirmed that these haplogroups encompass virtually all the mitochondrial lineages present in Europe and can be detected in both northern and southern European populations. We also discovered 2 restriction sites (-73 Alw441 and +75 SphI) that allow the detection of informative nucleotide changes in the second hypervariable segment of the control region, which help to detect the haplogroup identity of mtDNAs without requiring further DNA sequencing. Haplogroup H was the most common mtDNA lineage in this sample, reaching frequencies from about 40% in Corsican and Gallurese populations, to about 65% in the Barbagian population. Haplogroup V, possibly originating in the Iberian peninsula, was found only in the central Sardinian sample. Of the 5 Corsican mtDNAs belonging to the haplogroup T, 4 had a restriction fragment length polymorphism found only in this population. It seems that this mutation originated in Corsica and has had time to spread in the area, since the maternal grandmothers of the subjects came from different villages of the island. The sample from central Sardinia shows a remarkable discontinuity with those from the northern part of the island and from Corsica. Gallura and Corsica seem to have undergone a more recent peopling event, possibly related to the arrival of new mitochondrial variability from continental Italy, while Barbagia has apparently maintained more archaic haplotypes. PMID- 11048789 TI - HLA polymorphism and evaluation of European, African, and Amerindian contribution to the white and mulatto populations from Parana, Brazil. AB - Polymorphism of classical HLA-A, HLA-B, HLA-C, HLA-DR, and HLA-DQ genes differs greatly among populations, both in frequencies and in the presence of alleles and haplotypes particular to population groups, making these genes powerful tools for the study of origins of populations and their degree of admixture. Antigen, allele, and haplotype frequencies, together with linkage disequilibrium patterns, are reported for 2 populations in the southern Brazilian state of Parana, one of predominantly European ancestry (white), the other of predominantly African and European ancestry (mulatto). Genetic distance estimates between the 2 groups and other populations studied previously, and of degree of admixture, were performed. In accordance with phenotypic classification, the white population is of predominantly European origin (80.6%), with a smaller contribution of African (12.5%) and Amerindian (7.0%) genes. The mulatto population consists of African (49.5%) and European (41.8%) ancestry, with a smaller but significant contribution of Amerindian (8.7%) ancestry. On the basis of history and population genetics, there is controversy regarding the Amerindian contribution to Parana's gene pool. These results provide a better picture of Parana's ethnic constitution and on the Amerindian contribution to the white and mulatto populations. PMID- 11048790 TI - The genetic position of the autochthonous subpopulation of Northern Navarre (Spain) in relation to other basque subpopulations. A study based on GM and KM immunoglobulin allotypes. AB - GM and KM immunoglobulin (Ig) allotypes were tested in 118 autochthonous Basques from northern Navarre. The results are compared to those obtained for the same genetic markers in 6 other Basque subpopulations, 3 from Spain (Guipuzcoa, Vizcaya, and Alava) and 3 from France: Macaye, Saint-Jean Pied de Port, and Mauleon. The northern Navarrese appear genetically closer to the Alava and Saint Jean Pied de Port subpopulations. The Basques present 3 GM haplotypes that are uncommon in Caucasian populations, suggesting that they have not been completely isolated either from Asian or African populations. The GM*1,17 23' 10,11,13,15,16 north Asian haplotype was probably the first to be introduced into the Basque area. The GM*1,17 23' 5* haplotype, considered an African genetic marker although also detected in Central Asia, would have reached the Iberian Peninsula through consecutive historic migrations from North Africa. The rare haplotype GM*1,17 23 21,28 results probably from a genetic recombination or crossing-over between the 2 common haplotypes GM*1, 17 23' 21,28 and GM*3 23 5*. It is also found with a low frequency in other neighboring regions and countries; but the possibility of its having been introduced through the main passage connecting western France and Spain during the Roman Empire and Middle Ages cannot be ruled out. PMID- 11048791 TI - The relationship between maternal and offspring birth weights after maternal prenatal famine exposure: the Dutch Famine Birth Cohort Study. AB - We examined the impact of famine-induced changes in maternal birth weight (MBW) on the association between MBW and offspring birth weight (OBW). Women born before, during, and after the Dutch Famine of 1944-1945 were interviewed at ages 41 to 46 years. Women (n = 582) and their children (n = 1,111) were included in the analysis if both mother and child were singleton and the child was not delivered preterm. Mean birth weight (BW) of women with first-trimester exposure (n = 110) was 154 g higher (p = 0.008), and mean BW of women with third-trimester exposure (n = 138) was 251 g lower (p < 0.001) than mean BW of unexposed women (n = 302). First-born offspring of women with first-trimester exposure were 72 g heavier (95% confidence interval [CI], -57 to 201; p = 0.27), and offspring of women with third-trimester exposure were 43 g lighter (95% CI, -170 to 79; p = 0.47) than offspring of unexposed women. Among unexposed women, each 100 g increase in MBW was associated with 25 g (95% CI, 12 to 37) increase in OBW (adjusted for maternal age, smoking, weight, and height and offspring sex). This association was attenuated in famine-exposed women (first-trimester change in OBW = 20 g per 100 g MBW; 95% CI, -1 to 41; third-trimester change in OBW = 14 g per 100 g MBW; 95% CI, -9 to 37). When MBW and trimester of maternal famine exposure were considered in a joint model, there was no independent effect of trimester of maternal famine exposure on OBW. Associations were less consistent for later-born offspring. We conclude that maternal prenatal famine exposure does not affect the association between maternal and offspring BW. Trimester of exposure was not a determinant of OBW, other than through its effect on MBW. Nevertheless, acute famine may impact on second-generation BW distributions indirectly, through its effect on the distribution of MBW. PMID- 11048792 TI - Birth seasonality in the early Spanish-Mexican colonists of California (1769 1898). AB - The pattern of birth seasonality in California's early Spanish-Mexican colonists between 1769 and 1898 was reconstructed using genealogical data for progeny of 657 marriages. The monthly distribution of the 3,824 births in this sample shows a strong seasonal pattern, with spring and fall peaks (corresponding to peaks in conceptions during July and February) and a low point in October. This seasonal reproductive pattern is the result of a complicated set of interactions among environmental, physiological, and cultural variables. California's strongly developed winter rainfall pattern and the 19th-century agricultural cycle clearly influenced the seasonal pattern of marriages and births in this agrarian society. Several historical processes interacted with these environmental and economic factors to transform the seasonal birth pattern of the early colonists. Through time the birth pattern becomes less variable and the birth maximum shifts from spring to early winter. This appears to be, at least in part, a result of changes in labor patterns and an increase in average parity. These data suggest a multifactorial explanation for birth seasonality, in which the timing of conceptions and births is influenced by both environmental and socioeconomic factors. PMID- 11048793 TI - Trends in inbreeding, isonymy, and repeated pairs of surnames in the Valserine Valley, French Jura, 1763-1972. AB - The estimates of inbreeding derived from pedigrees and frequency of isonymous marriages (i.e., between persons of the same surname) are compared using genealogical and isonymic information from 4,899 marriages recorded between 1763 and 1972 in 4 rural villages of the French Jura region (a mountainous area near the Swiss border). Before the second half of the 20th century, the two kinds of estimates show a different temporal evolution. The mean inbreeding coefficient based on pedigrees increases between 1763 and 1852 and reaches a maximum between 1853 and 1882 (alpha = 0.0028), with a very low percentage (< 1%) the result of remote kinship. The mean inbreeding coefficient based on isonymy is always higher, with a maximum observed between 1793 and 1822 (F = 0.0200), and it remains roughly the same between 1763 and 1882 (F = 0.0150), with a high percentage resulting from a random component (Fr), a consequence of the small population size and genetic drift. After 1883, the 2 mean coefficients decrease. This discordance is largely explained by the poor quality, for the first periods, of the genealogical data base, which ignores the more remote links of kinship, justifying the use of the model of Crow and Mange (1965) to explore consanguinity during the more ancient periods. The temporal evolution of the repeated pairs of surnames index (RP) confirms the recent evolution of the marital structure of the valley. Moreover, it appears that isonymous marriages and repeated and unique pairs of surnames constitute 3 distinct matrimonial groups characterized by both a different mean coefficient of inbreeding (alpha) and a different rate of endogamy. PMID- 11048794 TI - A Pleistocene population X-plosion? PMID- 11048795 TI - Further data on the microsatellite locus D12S67 in worldwide populations: an unusual distribution of D12S67 alleles in Native Americans. AB - We report the frequencies of alleles at the microsatellite locus D12S67 in 2 widely separated ethnic groups of the world: 2 populations from Sulawesi, an island in the Indonesian archipelago, and 5 Native American tribes of Colombia, South America. The allele frequencies in the Minihasans and Torajans of Sulawesi are similar to each other (but the modal class allele is different) and in general agreement with those reported in mainland Asian groups, but different from both Europeans and Chinese Han of Taiwan. The 5 Native American tribes (Arsario, Kogui, Ijka, Wayuu, and Coreguaje) display different allele frequencies from those seen in Sulawesi populations, in other groups from Europe and mainland Asia, and in Chinese Han of Taiwan. Native Americans exhibit a bimodal distribution of alleles, unlike other groups, with significant differences among the tribes. The Arsario and Kogui have no admixture with Europeans or Africans and are the most distinctive, while the Wayuu have the most admixture and show most similarity to other groups. The data suggest that nonadmixed Native Americans may be quite distinctive with respect to this marker. The most common allele varies across the 5 tribes, from 249 base pairs to 261 base pairs. All samples exhibit Hardy-Weinberg genotype proportions; heterozygosities are lowest in the 2 nonadmixed Native American tribes. Examination of all the available data indicates that some east Asian and southeast Asian groups are characterized by a high frequency of smaller sized D12S67 alleles, while other populations have a greater proportion of the larger sized alleles. The cumulative, though still highly restricted, population data on locus D12S67 demonstrate that it may be of considerable value in anthropological genetic studies of ethnic groups. Data are required on Native Americans outside Colombia before this marker can be used in admixture studies of this group. PMID- 11048796 TI - Variability of the F13B locus in South American populations. AB - A population study for the F13B locus was carried out in a total of 396 South American individuals. The analysis comprised new data from 5 Amerindian populations, existing data from 3 Amerindian populations, and I urban sample from La Plata, Argentina. In both pooled Amerindian and La Plata samples, 6 alleles were found. The individual Amerindian samples showed a lower number of alleles, changes in modal alleles, and restricted variability. Interpopulation comparisons revealed significant differences among samples from distinct geographical regions. Differences among the groups were also corroborated by the F(ST) statistic. Data support the hypothesis that genetic drift and gene flow influence Amerindian differentiation. PMID- 11048797 TI - Mutations in beta-catenin and APC genes are uncommon in esophageal and esophagogastric junction adenocarcinomas. AB - Beta-catenin plays important roles in both intercellular adhesion and signal transduction. Mutations in the beta-catenin or adenomatous polyposis coli (APC) gene can alter the degradation of beta-catenin and cause aberrant accumulation of beta-catenin result in increased transcription of target genes. The dysregulated APC/beta-catenin pathway has been recently discovered as an important mechanism of tumorigenesis in various cancers, but its role in esophageal adenocarcinomas is not clear. Therefore, we studied the beta-catenin gene mutation, allelic loss of chromosome 5q, and APC gene mutation in esophageal and esophagogastric junction adenocarcinomas. Two (2%) somatic mutations in exon 3 of the beta catenin gene, encompassing the region for glycogen synthase kinase-3beta phosphorylation, were detected from 109 adenocarcinomas. Chromosomal allelic loss on 5q was frequent in 45.3% (44/97) of tumors. Only one missense mutation in the mutation cluster region of the APC gene was detected from 38 esophageal and esophagogastric junction adenocarcinomas with the 5q allelic loss. Our results based on partial screening mutational analyses indicate that mutations of APC/beta-catenin pathway, unlike in colorectal carcinoma, involve only a small subset of esophageal and esophagogastric junction adenocarcinoma. PMID- 11048798 TI - Immunohistochemical and in situ hybridization analyses of midkine expression in thyroid papillary carcinoma. AB - Midkine (MK) is a novel heparin-binding growth factor whose gene has been identified in embryonal carcinoma cells in early stages of retinoic acid-induced differentiation. We immunohistochemically examined 90 thyroid papillary carcinomas (85 invasive type and five encapsulated type), using a rat IgG2a monoclonal antibody against the carboxyl terminal region of human MK in archival paraffin sections. The thyroid tumors exhibited an intense reaction in the cytoplasm. Most of the papillary carcinomas (77/90), had tumor cells that expressed MK. These were classified into the following two types: invasive type (76/85) and encapsulated type (1/5). Notably, the intensity of MK was stronger at the invading border area of the tumors than in the center. In tissues adjacent to the cancer tissues, normal follicular epithelial cells expressed MK very faintly or not at all. The in situ hybridization analysis revealed that the signals of MK transcripts were found in the cytoplasm of the cancer cells. In the noncancerous follicular epithelial cells adjacent to neoplasm the signals of MK transcripts were detected very weakly or not at all. The distribution and localization of the MK-transcript signals determined by in situ hybridization analysis were similar to those obtained by immunohistochemical analysis. We conclude that thyroid papillary carcinoma strongly expresses MK protein and messenger RNA, and that this overexpression may relate to the development and invasion of these carcinomas. PMID- 11048799 TI - Mutational analysis of the CTNNB1 and APC genes in uterine endometrioid carcinoma. AB - Despite recent studies, the molecular genetic events responsible for the development of uterine endometrioid carcinoma (UEC) remain incompletely characterized. Mutations in the beta-catenin (CTNNB1) gene have been recently reported in a small percentage of UECs and in the endometrioid variant of ovarian carcinoma suggesting that the Wnt signal transduction pathway is involved in the development of female genital tract tumors with endometrioid morphology. The Wnt pathway is a critical pathway in the development of colorectal cancer (CRC) with mutations occurring in the beta-catenin (CTNNB1) or adenomatous polyposis coli (APC) genes in 10 to 15% and 85% of cases, respectively. Because UEC and CRC share other molecular genetic alterations and histologic features and previous studies of UEC have not reported an analysis of the APC gene, we chose to further elucidate the role of the Wnt pathway in UEC. To this end, we analyzed 32 cases of UEC for mutations of the CTNNB1 and APC genes. Mutations of CTNNB1 were present in six of 32 (18%) cases: four grade 1 carcinomas, one grade 2, and one grade 3 carcinoma. Five missense mutations were identified, three involving Ser/Thr phosphorylation sites and two adjacent to a Ser phosphorylation site. One case contained a deletion encompassing codons 34 to 37, which includes a Ser phosphorylation site. No mutations resulting in truncation of the APC protein were found. Our results support a role for the Wnt signaling pathway via mutation of CTNNB1, but not APC, in the development of a subset of UECs. PMID- 11048800 TI - Gastric cardia intestinal metaplasia: biopsy follow-up of 85 patients. AB - BACKGROUND: Gastric cardia intestinal metaplasia (CIM), denoted by goblet cells is common. The frequency of persistent CIM is unknown. METHODS: 85 patients with CIM and follow-up endoscopies were prospectively identified during the time period of 10/6/94-12/21/97. The presence of goblet cells was the defining feature of CIM, other metaplastic cell types were not evaluated. AU 85 patients initially had biopsies that straddled the squamocolumnar junction (SCJ) showed CIM, an otherwise normal proximal stomach, lower esophagus, and squamocolumnar junction. The SCJ lay within the 2 cm of mucosa immediately proximal to the uppermost gastric fold and overlaid the junction of the tubular esophagus and the saccular dilatation of the stomach in all patients. The patients underwent endoscopy for many reasons. They were randomly identified based on the absence of a hiatal hernia and the presence of CIM. RESULTS: Ten of the 85 patients had CIM on repeat biopsy. Among patients with no CIM in the first repeat endoscopy, the degree of cardia inflammation decreased between the initial and first repeat endoscopy, whereas there was no change in the amount of inflammation among patients who had CIM in the first repeat endoscopy. The changes in mean inflammation score was significantly different between the two groups (P = .024). Twenty-two patients underwent a second repeat endoscopy and five had a third repeat endoscopy. Including all follow-up biopsies, six of the 85 patients (7%) had CIM. Four patients who did not have CIM on initial repeat endoscopy had CIM on their second repeat endoscopy, probably reflecting sampling issues. None of the biopsies had dysplasia. CONCLUSIONS: Cardia inflammation is a stimulus for cardia intestinal metaplasia, and a reduction in inflammation may allow the metaplastic mucosa to revert to normal. PMID- 11048801 TI - Cytogenetic, clinical, and morphologic correlations in 78 cases of fibromatosis: a report from the CHAMP Study Group. CHromosomes And Morphology. AB - Whether fibromatoses are neoplastic or reactive lesions has long been controversial and the relationship, if any, between the superficial and deep forms (desmoid tumors) are poorly understood. Clinical, pathologic, and cytogenetic data of 78 cases of fibromatosis were analyzed and correlated with each other. The results demonstrate that clonal chromosome aberrations are a common feature of this entity, being present in 46% of desmoid tumors, although less frequent in the superficial types (10%). In the deep-seated extra-abdominal fibromatoses, trisomies 8 and 20 and loss of 5q material were the only recurrent features. No correlation between +8 and local recurrence was found. Our findings provide additional evidence for the neoplastic nature of fibromatoses. PMID- 11048803 TI - DNA copy number changes in epithelioid sarcoma and its variants: a comparative genomic hybridization study. AB - Epithelioid sarcoma is a distinctive, rare soft tissue sarcoma that typically involves the distal extremities in young adults, and shows epithelioid morphology and immunohistochemical markers of epithelial differentiation. The genetic background of epithelioid sarcoma is poorly understood, and knowledge of it could give insights into the pathogenesis of this tumor and its possible relationship with other malignant tumors. In this study, we analyzed DNA copy number changes in 30 epithelioid sarcomas by comparative genomic hybridization. DNA was extracted from microdissected samples of formaldehyde-fixed and paraffin-embedded tumors with a minimum of 60% of tumor cells in each sample. Sixteen tumors (53%) showed DNA copy number changes at one to six different genomic sites. The majority of the changes were gains, seen in 14 tumors, whereas 10 tumors showed losses. The most common recurrent gains were at 11q13 (five cases), 1q21-q23 (four cases), 6p21.3 (three cases), and 9q31-qter (three cases). High-level amplifications were detected once in 6p21.3-p21.1 and once in 9q32-qter. Recurrent losses were seen at 9pter-p23 (three cases), 13q22-q32 (three cases), 1p13-p22 (two cases), 3p12-p14 (two cases), 4q13-q33 (two cases), 9p21 (two cases), and 13q32-qter (two cases). The most common recurrent gain at 11q13 was seen in both classic cases and angiomatoid and rhabdoid variants supporting the relationship of these variants with the classic epithelioid sarcoma. Expression of cyclin D1 gene, located in 11q13, was immunohistochemically detected in nine of 15 cases including three of five cases with gain of 11q13, suggesting its involvement in epithelioid sarcoma. The observed comparative genomic hybridization changes give targets for future genetic studies on epithelioid sarcoma. PMID- 11048802 TI - Intraarticular osteoid osteoma associated with synovitis: a possible role of cyclooxygenase-2 expression by osteoblasts in the nidus. AB - To clarify the condition of development of synovitis associated with intraarticular osteoid osteoma (OO), expressions of cyclooxygenase-2 (COX-2) protein and its messenger ribonucleic acid were investigated both in the nidus and the synovial tissue using immunohistochemical and reverse transcription polymerase chain reaction analyses. Diffuse and strong COX-2 immunoreactivity was found in osteoblast-like tumor cells in the nidus of all six cases of OO (three of six cases were intraarticular OO associated with synovitis) and one case of osteoblastoma associated with synovitis. Expression of COX-2 messenger ribonucleic acid was demonstrated in one case of OO associated with synovitis, and was higher in the nidus than that in the inflamed synovial tissue. However, there were no significant difference between the nidus and synovium in the expression of cytosolic phospholipase A2, one of the enzymes involved in arachidonic acid metabolism, and inflammatory cytokines such as interleukin-1beta and tumor necrosis factor alpha. Finally, as there was only one case in which the examinations of gene expression were performed, no definitive overall conclusions could be reached; rather it is suggested that COX-2 expressed primarily by osteoblasts in the nidus of intraarticular OO may play a role in activating the pathway of arachidonic acid metabolism, resulting in synovitis of the involved joint. PMID- 11048804 TI - Expression of pituitary homeo box 1 (Ptx1) in human non-neoplastic pituitaries and pituitary adenomas. AB - We investigated the localization of pituitary homeo box 1 (Ptx1) protein in five human non-neoplastic pituitaries and 73 of all types of pituitary adenomas using immunohistochemistry, and the expression of Ptx1 messenger RNA (mRNA) in 18 representative pituitary adenomas using the reverse transcriptase polymerase chain reaction (RT-PCR) technique. By immunohistochemical analysis, Ptx1 protein was extensively detected in the nuclei of normal human pituitary cells. Ptx1 was detected in 10/14 (71.4%) of growth hormone (GH)-secreting adenomas, 12/12 (100%) of prolactin (PRL)-secreting adenomas, 18/20 (90%) of adrenocorticotropic hormone (ACTH)-secreting adenomas, 6/7 (85.7%) of thyroid-stimulating hormone (TSH) secreting adenomas, and 17/20 (85%) of clinically non-functioning adenomas, including 9/10 (90%) of gonadotropin-subunit-positive adenomas. Thus, there was no relationship between Ptx1 expression and a particular type of pituitary adenomas. By RT-PCR analysis, Ptx1 mRNA was expressed in all 18 cases of pituitary adenomas, including two cases negative for Ptx1 protein by immunohistochemistry. These results suggested that Ptx1 may be an universal transcription factor in both neoplastic and non-neoplastic conditions in human pituitaries. The synergistic action with other transcription factors may be speculated to determine the specific production of the anterior pituitary hormones. PMID- 11048805 TI - Vinculin: its possible use as a marker of normal collecting ducts and renal neoplasms with collecting duct system phenotype. AB - Vinculin is a cytoskeletal protein associated with membrane actin-filament attachment sites of cell-cell and cell-matrix adherens-type junctions. In this article, we examine the expression of vinculin to elucidate its role in human renal neoplasms. We reviewed surgically resected specimens and selected available tissue from 79 renal tumors in 78 patients. There were 55 men and 23 women. Their mean age was 61 years and the mean size of the renal tumors was 6.1 cm. All renal tumors were examined by immunohistochemistry using a monoclonal antibody against vinculin. Overall, 17 (21.5%) renal tumor samples reacted with vinculin. The positive ratio in various types of renal tumors was as follows: conventional-type (clear cell), 0/54; papillary-type, 5/12; chromophobe-type, 5/5; sarcomatoid type, 3/4; collecting duct carcinoma, 3/3; and oncocytoma, 1/1. The positive rate of conventional-type renal cell carcinomas (RCCs) is significantly different from that of other renal tumors (P < .01). Normal kidney, conventional, and papillary type RCCs exhibited positive signals in Western blot analysis. These results suggest that vinculin may serve as a useful marker of renal neoplasms with collecting duct system phenotype such as chromophobe-type RCC. PMID- 11048806 TI - Expression of Bax, Bcl-2, and P53 in progressive multifocal leukoencephalopathy. AB - It has been shown in vitro that JC viral protein can form a complex with wild type p53 protein, which is a key regulator of both cell proliferation and cell death. Cellular factors, Bax and Bcl-2, are two essential downstream elements involved in p53-dependent apoptosis. To determine whether association of JC virus with p53 protein affects the expression of Bax and Bcl-2 in viral-infected cells in progressive multifocal leukoencephalopathy (PML), we studied the expression of Bax, Bcl-2, and p53 in 14 cases from 13 PML patients by using paraffin immunohistochemistry. Seven of 13 patients were known to be HIV positive. Overexpression of p53 was found in viral-infected oligodendrocytes and some astrocytes in all 14 cases. Intense immunostaining of Bax was strongly expressed in viral-infected oligodendrocytes and astrocytes. Bax immunostaining was also found in macrophages in the demyelinating lesions. Bcl-2 was not detected in viral-infected glial cells. The expression pattern of Bax positive/Bcl-2 negative in viral-infected glial cells suggests that the oligodendrocyte may be undergoing apoptosis which may in turn contribute to the demyelinating process in PML. The coexpression of p53 and Bax in the infected glial cells suggests that p53 detected by immunohistochemistry may still maintain its wild-type function. PMID- 11048807 TI - CD44H and CD44V6 expression in different subtypes of Hodgkin lymphoma. AB - CD44 is a broadly distributed family of cell surface glycoproteins. The expression of CD44H has been documented in both Hodgkin lymphoma and non-Hodgkin lymphoma. CD44V6 has been associated with more aggressive behavior in non-Hodgkin lymphoma, but such a correlation has not been established in Hodgkin lymphoma. In addition, the utility of CD44 and CD44V6 in the subclassification of Hodgkin lymphoma in paraffin-embedded tissues has not previously been evaluated. The current study included formalin- or methacarn-fixed, paraffin-embedded tissue specimens from 42 patients with Hodgkin lymphoma (25 nodular sclerosis, three interfollicular, four lymphocyte-rich classic Hodgkin, six lymphocyte predominant, and four mixed cellularity). The clinical stage of the study population at initial presentation ranged from stage IA to IVB. Evaluation of CD44H and CD44V6 (Novocastra) was performed by ABC immunoperoxidase technique after heat-induced epitope retrieval. In the six cases of lymphocyte predominant Hodgkin, the neoplastic cells lacked reactivity with CD44H reminiscent of their normal germinal center counterparts. On the other hand, classic Hodgkin lymphoma showed variable membranous and Golgi reactivity in the neoplastic cells in all cases irrespective of disease stage at presentation. In all cases, the neoplastic cells lacked reactivity with CD44V6 except for three one lymphocyte-predominant, one interfollicular, and one nodular sclerosis), all of which represented recurrent cases. In conclusion, CD44 evaluation is useful in the distinction between lymphocyte predominant and classic Hodgkin lymphoma. The presence of CD44H expression has no relation to the clinical stage of the disease at presentation or recurrence. CD44V6 is detected in a minority of cases irrespective of the histologic subtype and its presence may be associated with recurrence. There was no correlation between disease stage at presentation and the expression of CD44V6. PMID- 11048808 TI - Tonsillar lymphangiomatous polyps: a clinicopathologic series of 26 cases. AB - BACKGROUND: Lymphangiomatous polyps are uncommon benign tumors of the tonsils. METHODS: Twenty-six cases of lymphangiomatous polyps diagnosed between 1980 and 1999 were retrieved from the files of the Otorhinolaryngic-Head and Neck Tumor Registry of the Armed Forces Institute of Pathology. Hematoxylin and eosin stained slides were reviewed to characterize the histologic features of these tumors. Immunohistochemical stains were performed on 15 cases. Clinical follow-up data were obtained. RESULTS: The patients included 13 males and 13 females, ages 3 to 63 years (mean, 25.2 years). Patients experienced dysphagia, sore throat, and the sensation of a mass in the throat. Symptoms were present from a few weeks to years. The tonsillar masses were unilateral in all cases. Clinically, the lesions were frequently mistaken for a neoplasm (n = 18 patients). Grossly, all of the lesions were polypoid and measured 0.5 to 3.8 cm (mean, 1.6 cm). Histologically, the polyps were covered by squamous epithelium showing variable epithelial hyperplasia, dyskeratosis, and lymphocytic epitheliotropism. The masses showed a characteristic submucosal proliferation of small to medium-sized, endothelial-lined, lymph-vascular channels lacking features of malignancy. Collagen, smooth muscle, and adipose tissue were present in the stroma. Intravascular proteinaceous fluid and lymphocytes were noted. Immunohistochemical findings confirmed the endothelial origin of the vascular proliferation and a mixed lymphoid population. The differential diagnosis included fibroepithelial polyp, lymphangioma, juvenile angiofibroma, and squamous papilloma. In all patients with follow-up, complete surgical excision was curative (mean follow-up, 5.4 years; range, 1 mo to 14 years). CONCLUSIONS: We detail the clinical and pathologic features of tonsillar lymphangiomatous polyps. These tumors are uncommon and may clinically be mistaken for a malignant neoplasm. The characteristic histologic features should allow for its correct diagnosis and differentiation from similar appearing tonsillar lesions. PMID- 11048809 TI - Immunohistochemical spectrum of GISTs at different sites and their differential diagnosis with a reference to CD117 (KIT). AB - Gastrointestinal (GI) stromal tumor (GIST) is the designation for the major subset of GI mesenchymal tumors and encompasses most tumors previously classified as GI smooth muscle tumors. Although GISTs typically express CD117 (KIT), often express CD34, and sometimes express alpha-smooth muscle actin (SMA), the relative frequency of these markers has not been characterized in large series of GISTs of different sites, and the CD117 expression has not been fully characterized in intra-abdominal tumors. In this study, we immunohistochemically analyzed 292 GISTs throughout the GI tract, including omentum and mesentery, and compared the immunoreactivities with 211 other tumors that may enter in the differential diagnosis. GISTs were defined in this study as CD117-positive primary spindied or epithelioid mesenchymal tumors of the GI tract, omentum, or mesentery. The CD34 positivity of GISTs varied from 47% in small bowel to 96 to 100% in rectum and esophagus, whereas SMA expression showed the opposite patterns and was most frequent in the GISTs of small bowel (47%) and rarest in the GISTs of rectum and esophagus (10-13%). Desmin was seen only occasionally. S100 positivity was rare but was seen most frequently in small intestinal GISTs (15%). True leiomyomas from esophagus, muscularis mucosae of colorectum, and pericolic leiomyomas similar to uterine leiomyomas were negative for CD117 and CD34 and positive for SMA and desmin (46 of 46). Inflammatory fibroid polyps of stomach and small intestine were negative for CD117 but were often positive for CD34 (6 of 8) and variable for SMA (3 of 8). Inflammatory myofibroblastic tumors involving gastric or colonic wall were negative for CD117 but some showed CD117-positive endothelia. GI schwannomas were all negative for CD117 and positive for S100 protein (11 of 11). Extremely focal CD117 positivity was seen in the neoplastic cells of some retroperitoneal leiomyosarcomas and liposarcomas. Among other CD117 positive tumors were intestinal metastatic melanomas (8 of 11) and extraskeletal Ewing's sarcomas (5 of 11), two of which were abdominal. In conclusion, strong CD117 expression defines most primary GI mesenchymal tumors as GISTs, which show different patterns for CD34 and SMA in various parts of the GI tract. Some unrelated CD117-positive tumors (melanomas, Ewing's sarcomas) should not be confused with GISTs. PMID- 11048810 TI - Anaplastic large cell lymphomas presented as bone lesions: a clinicopathologic study of six cases and review of the literature. AB - Non-Hodgkin's lymphomas uncommonly present as bone lesions. Most of these tumors are diffuse large B-cell lymphomas. Anaplastic large cell lymphoma (ALCL) presented as bone lesions is exceedingly rare. In this study, we describe six cases of ALCL that presented as solitary or multiple bone lesions. The average patient age was 33 years (range, 4 to 63 years) and the male to female ratio was 2:1. Fever and localized bone pain were the most frequent presenting symptoms. Radiologic examinations revealed osteolytic lesions in all cases, with three (50%) being multiple lesions and five (83%) involving the axial bones. All patients were initially assessed to have only bone involvement. Staging studies revealed mild cervical lymphadenopathy in one patient and no evidence of extraskeletal disease in the other five patients. Histologically, there was diffuse infiltration of one or more bones by large pleomorphic lymphoma cells. Immunohistochemical studies showed all six neoplasms were positive for CD30, EMA, and granzyme B. One case was of T-cell lineage, positive for CD3. One case was positive for the T-cell-associated antigen CD4. The remaining four cases were of null-cell type. In-situ hybridization for EBV was performed in five cases; all were negative. Despite the relatively low International Prognostic Index (IPI) of these patients (mean, 1.67; range, 1 to 3), the overall prognosis was relatively poor: three of six died of disease within 2 years of diagnosis, and two of six were alive with evidence of disease (follow-up, 6 mo to 2 years). Thus, compared to their nodal counterparts, ALCLs that present as bone lesions are distinguished by their uniform expression of EMA and granzyme B, and a relatively poor clinical outcome. Our results also suggest that ALK-1 expression in this clinical setting is not a favorable prognostic indicator. PMID- 11048811 TI - Spindle epithelial tumor with thymus-like differentiation (SETTLE): a distinctive malignant thyroid neoplasm with significant metastatic potential. AB - Spindle epithelial tumor with thymus-like differentiation (SETTLE) is a very rare tumor of the thyroid believed to be derived from branchial pouch or thymic remnants and showing primitive thymic differentiation. Although this tumor is prone to develop delayed blood-borne metastases, the metastatic risk is unclear because the case reports in the recent literature had very short follow-up periods. We report one case, the oldest patient reported so far, who had been followed up until death. The 59-year-old man had an enlarged thyroid for all of his adult life, and presented with recent rapid enlargement of the thyroid. Neck exploration revealed a hard tumor in the right lobe, with adhesion to sternothyroid muscle. Histologic examination showed an invasive biphasic neoplasm traversed by sclerotic septa. Tight to loose fascicles of bland-looking spindly cells were intimately intermingled with tubulopapillary structures, diagnostic of the SETTLE. This patient developed left pulmonary metastases at 2 years and subsequently developed bilateral pulmonary and widespread metastases. He died 8 years after initial presentation. This case illustrates the protracted clinical course of the tumor, and survival for many years despite the occurrence of metastases. Review of the literature shows that SETTLE occurs predominantly in young patients with a median age of 15 years and male predominance. There is a significant metastatic rate of 71% for patients with more than 5 years of follow up in spite of the otherwise indolent nature of the tumor. PMID- 11048812 TI - Epithelioid leiomyosarcoma in a non-immunocompromised infant: additional differential diagnosis of pediatric "round cell tumors". AB - We report an 18-month-old Japanese girl with purely epithelioid leiomyosarcoma presenting as a huge intraabdominal mass. The patient had been well from birth and had shown no signs of immunodeficiency. She was negative for human immunodeficiency virus. Blood examination revealed elevated serum neuron specific enolase (NSE). Histologically, the tumor was comprised of solid growths of round or polygonal cells with vesicular nuclei and often vacuolated cytoplasm rich in glycogen. The tumor cells were positive for vimentin, NSE, and MIC2, and were negative for desmin and neurofilament. The age, clinical presentation, and histologic findings mostly favored Ewing's sarcoma/primitive neuroectodermal tumor. Silver stain, however, demonstrated well-developed reticulin fibers often outlining individual tumor cells. An expanded panel of immunostains showed that the tumor cells were intensely positive for smooth muscle actin, and ultrastructural study revealed abundant fine cytoplasmic filaments with focal subsarcolemmal densities, various amounts of glycogen, and irregularly arranged, thick basal lamina. The diagnosis of epithelioid leiomyosarcoma was made. Following reduction in tumor size by chemotherapy, the serum NSE level was normalized. From the surgical finding, the primary site was presumed to be the urachus or the urinary bladder dome. Although extremely rare, epithelioid leiomyosarcoma should be added in the list of differential diagnoses of pediatric "round cell tumors." PMID- 11048813 TI - Prodromal schizophrenia and atypical antipsychotic treatment. AB - Early recognition and intervention in psychosis is the focus of more intensive research. In this paper, we critically review the ideas that have emerged in this field. We also propose a model or hypothesis for testing in the prodromal phase of schizophrenia. Attention to practical and ethical issues, particularly with the use of atypical antipsychotics in one arm of the protocol, is addressed. Studies by Yung and Falloon describe prodromal intervention with psychosocial strategies and time-limited low potency neuroleptics, respectively, that suggest benefits of such a model. Although we have respect for the DSM system, this paper is written more from a Bleulerian than Kraepelinian perspective in that we emphasize affective, cognitive, and negative symptoms in addition to positive symptoms. The paper recognizes the strong conceptual disagreements implicit in this area stemming not only from Kraepelin and Bleuler but work from the 1930s by Cameron. The clinical research advocated is timely in that the atypicals are more congruent to the Bleulerian conception with a neurodevelopmental hypothesis of schizophrenia. We also have exciting new imaging and genetic technologies to refine our concepts of schizophrenia and its prodromal and premorbid phases. PMID- 11048814 TI - Assessing positive and negative symptoms in outpatients with schizophrenia and mood disorders. AB - The Positive and Negative Syndrome Scale (PANSS) was developed to assess symptomatology in inpatients with schizophrenia. We examined its reliability and validity among outpatients with schizophrenia (N = 75) and mood disorders (N = 61). Because the hypothesized three-scale structure of the PANSS has not been supported by existing research, we also examined its factor structure. Interrater reliability for individual items and the positive and negative scales was demonstrated. Evidence supported the internal consistency of each scale in the overall sample and the schizophrenia group but was mixed among those with mood disorders. PANSS scores were higher in the schizophrenia group. These scores, in turn, were lower than those generally reported among inpatients with schizophrenia, providing known-groups validity evidence. Four of five factors were similar to those reported among inpatients with schizophrenia. Together, these results support the use of the PANSS among outpatients and reinforce existing support for assessing positive and negative symptoms in mood disorders. PMID- 11048815 TI - Distinguishing between overlapping somatic symptoms of depression and HIV disease in people living with HIV-AIDS. AB - HIV-AIDS is a prevalent medical diagnosis in U.S. cities, and symptoms of depression are common in persons with HIV infection. This study examined the effects of overlapping symptoms of HIV disease and somatic depression that can inflate scores on the Beck Depression Inventory (BDI) and the Centers for Epidemiological Studies Depression Scale (CESD). Results from 357 HIV positive men and women identified discrete subsets of depression symptoms that correspond with symptoms of HIV infection. Removing somatic subsets of depression symptoms improved the clinical utility of the BDI and CESD. Clearer symptom separation occurred with the BDI than the CESD, but the CESD may be more sensitive than the BDI to depression associated with progression of HIV disease. Findings suggest that depression scales that include somatic symptoms will inflate depression scores in people living with HIV infection, and available methods for distinguishing overlapping symptoms should be employed when assessing people living with HIV infection. PMID- 11048816 TI - Associations among symptoms of depression and posttraumatic stress disorder and self-reported health in sexually assaulted women. AB - Symptoms of depression and posttraumatic stress disorder (PTSD) were examined for their association with health status in a sample of sexual assault victims. Hypotheses were that symptoms of each disorder would account for unique variance in health status among individuals exposed to traumatic stressors. Fifty-seven sexually assaulted college women were assessed for prior victimization history, assault characteristics, and depressive and PTSD symptoms. When prior history of sexual victimization, assault severity, and physical reactions during the assault were controlled, hierarchical multiple regression models indicated that symptoms of PTSD and depression were significantly associated with global health perceptions and severity of self-reported health symptoms. Only PTSD symptoms were significantly associated with reproductive health symptoms. The results suggest that both symptoms of PTSD and depression account for the relationship between exposure and health impairment among sexual assault victims. PMID- 11048817 TI - Women in treatment: within-gender differences in the clinical presentation of opioid-dependent women. AB - Despite consistent evidence of gender differences in the nature of drug dependence, there has been little consideration of within-gender differences in the clinical presentation of drug-abusing women. In this study, cluster analysis and standardized ratings obtained from 153 women seeking methadone maintenance treatment were used to define four groups of women with different profiles of problem severity. The four clusters were characterized as Unemployed, Medically Ill, Psychiatrically Distressed, and Higher Functioning. When the validity of this four-cluster solution was examined, there were significant differences in the ethnic composition of the four groups, and the four clusters differed in terms of a) psychiatric status, b) medical status, c) vocational-educational history, d) lifetime history of maltreatment, and e) perception of social support available from friends and family. The findings suggest that, although understanding of gender differences cannot be ignored, understanding of ways women differ from one another may be as important in the development of gender sensitive treatment programs. PMID- 11048818 TI - Close relationships and social competence of hospitalized and nonhospitalized adolescents. AB - This study was designed to examine the social relationships of adolescents with severe disorders. The study sample consisted of 33 adolescent inpatients in a psychiatric unit, as well as 33 nonhospitalized adolescents. Participants completed the Network of Relationship Inventory for an assessment of the quality of their relationships with mother, father, close friend, and other significant adults. In addition, friendship intimacy was assessed. Results showed nonhospitalized adolescents describing a close relationship with their parents, as well as with their close friends. Their relationships with their friends were mature. Hospitalized adolescents described a less close relationship with their parents, whereas they were close to and valued their relationships with their friends and other significant adults. Though both groups described similar levels of emotional closeness, hospitalized adolescents tended more to exert control or to evince a penchant for similarity to the other in their friendships. Results are discussed within the framework of adolescent social tasks. PMID- 11048819 TI - Social sharing of Gulf War experiences: association with trauma-related psychological symptoms. AB - It commonly is believed that talking with family and friends (social sharing) about stressful or traumatic experiences can be therapeutic with regard to stress related psychological symptoms. Two years after serving in the Gulf War, 58 National Guard Reservists completed the Mississippi Posttraumatic Stress Disorder Scale (PTSD), the Brief Symptom Inventory, and a measure of social sharing that asked how much they had talked to family and friends about their experiences in the Gulf during the 2-year period since returning from the war. Subjects had a broad range of Mississippi PTSD scores. Six subjects met Mississippi criteria for PTSD. Degree of talking to family and friends about Gulf War experiences did not account for a significant portion of the variance in the prediction of PTSD symptoms but did significantly contribute to prediction of scores for interpersonal sensitivity, depression, and psychoticism. Thus, degree of talking with family and friends was not found to be related to PTSD symptoms, although it may have influenced some symptoms of general psychopathology, such as depression, that are not specific to PTSD. PMID- 11048820 TI - Longitudinal analysis of development among single and nonsingle children in Nanjing, China: ten-year follow-up study. AB - In order to examine the impact of "one-child-per-couple" family planning policy on child development, a longitudinal study of a group of children in Nanjing, China, was carried out between 1984 and 1995. The same cohort of children was examined at four stages of development: preschool, early school, preadolescence, and adolescence. Achenbach's Child Behavior Checklist was used to measure the child's behavior. A total of 274 children were surveyed in all four stages. The total behavior problem scores obtained at different stages of development were compared among boys and girls. The results showed that the total score declined significantly in association with the time. As the child grew older, the problems score decreased significantly. This maturation effect was seen for both boys and girls. On average, boys' scores dropped from 21 to 11, whereas girls' scores went from 19.5 to 12. In comparing single with nonsingle boys, there were no significant differences throughout the four stages of development. However, girls who were single children, in contrast to girls with siblings, had significantly higher scores of total behavior problems at each stage of development. These findings illustrate that the longitudinal effect of being a "single-child" is similar to other studies, indicating a minimal impact of family policy. However, the effects were more prominent among girls. This phenomenon can be explained by parents' different attitude toward single-children because of gender difference. PMID- 11048821 TI - Clinical presentations of posttraumatic stress disorder across trauma populations: a comparison of MMPI-2 profiles of combat veterans and adult survivors of child sexual abuse. AB - This investigation examined differences in symptom patterns of two different trauma samples using the Minnesota Multiphasic Personality Inventory-2 (MMPI-2). MMPI-2s of 122 male combat veterans seeking outpatient treatment for combat related PTSD were compared with those of 64 PTSD-diagnosed adults seeking outpatient treatment for the effects of child sexual abuse (CSA). We examined variables related to degree of health concerns, depression, somatization, anger and hostility, masculine-feminine traits, paranoid ideation, anxiety, difficulties thinking and concentrating, elevated mood, and social introversion, as well as test-taking attitude. MANOVAs revealed between-group differences on several variables. However, when analyses controlled for the effect of age, nearly all differences disappeared; the only remaining difference was in a scale measuring anger. Thus, it appears CSA survivors and combat veterans are much more similar than different in their clinical presentation on the MMPI-2. Conceptual issues in the assessment of PTSD are discussed. PMID- 11048822 TI - Independent relationships among outcomes and their predictors in subjects with panic attacks. PMID- 11048823 TI - Framing clinical questions. PMID- 11048824 TI - A case for lipid-lowering? PMID- 11048825 TI - Rapid-acting insulin analogues: when randomized clinical trial results do not help. PMID- 11048826 TI - HOPE for all people with diabetes? Heart Outcomes Prevention Evaluation. PMID- 11048827 TI - Effect of low-sodium diet on uteroplacental circulation. AB - OBJECTIVE: To study the influence of chronic dietary sodium restriction on uteroplacental circulation. METHODS: In a randomized trial, Doppler flow velocity waveforms of the uterine and umbilical artery were studied at monthly intervals during pregnancy in 59 women on a low-sodium diet and in 68 controls. RESULTS: Pulsatility index (PI), resistance index (RI), and A/B ratio of the uterine artery were significantly lower during sodium restriction, whereas PI, RI, and A/B ratio of the umbilical artery were significantly higher. CONCLUSIONS: The lower resistance indices of the uterine artery during sodium restriction might reflect an increase in pulse pressure/impedance ratio as a result of activation of the renin-angiotensin system. The increase in umbilical artery resistance indices supports the hypothesis that fetal circulation might be altered by chronic dietary sodium restriction. PMID- 11048828 TI - Use of maternal plasma level of zinc-coproporphyrin in the prediction of intrauterine passage of meconium: a pilot study. AB - OBJECTIVE: To evaluate the usefulness of maternal plasma zinc-coproporphyrin (ZCP) level as a marker for intrauterine passage of meconium. METHODS: A pilot study consisting of 10 pregnancies with meconium-stained amniotic fluid and 10 pregnancies with clear amniotic fluid was used. The corresponding plasma and amniotic fluid levels of ZCP were measured using spectrofluorometry. ZCP levels in plasma and amniotic fluid were compared between the two groups and the relation between plasma and amniotic fluid ZCP levels in the clear and meconium stained groups was assessed using Spearman rank-order correlation. RESULTS: Mean amniotic fluid ZCP was significantly higher in the meconium-stained amniotic fluid as compared to the clear amniotic fluid group. Although mean plasma ZCP levels were higher in the meconium-stained amniotic fluid group, this difference was not statistically significant. There was no significant correlation between plasma ZCP levels and amniotic fluid ZCP, but we could categorize patients according to plasma ZCP levels into four categories with different risks for having meconium-stained amniotic fluid. CONCLUSIONS: Plasma ZCP might be a promising test for prediction of intrauterine passage of meconium in high-risk patients if confirmed by larger studies. The implications of this prediction on management remain unknown. PMID- 11048829 TI - Intravenous glucose infusion in labor does not affect maternal and fetal acid base balance. AB - OBJECTIVE: This prospective randomized trial compares the effects of a 5% glucose solution or no infusion during labor on glucose levels, pH, pO2, pCO2, and base excess (BE) of normal pregnant women in early labor and at delivery, and on fetal cord blood. METHODS: Forty-three women were randomized to glucose infusion and 38 were controls. RESULTS: Starting glucose levels were independent from the fasting state. When no glucose supplementation was given, the labor itself was associated with a reduction of mean pH (from 7.42 to 7.36, P = 0.00001), mean pCO2 (from 25.7 to 24.4 mm Hg, P = 0.04), and mean BE (from -6.3 to -9.8 mEq/L, P = 0.00001), and an increase of capillary glucose (from a mean of 83 to 105 mg/dL, P = 0.00001). Infusions of glucose did not significantly alter maternal acid-base balance at delivery. pH, PO2, pCO2, and BE were similar in arterial and venous cord blood of both groups. No variables correlated with cord blood glucose levels or with glucose vein-artery difference. CONCLUSIONS: We conclude that a 5% glucose infusion does not significantly reduce maternal acidemia associated with vaginal delivery and therefore its use cannot be recommended, since maternal glucose is largely available during labor. Intrapartum glucose infusions do not alter the acid-base balance, when the fetus is well oxygenated. PMID- 11048830 TI - Umbilical cord serum levels of thromboxane B2 in term infants of women who participated in a placebo-controlled trial of low-dose aspirin. AB - OBJECTIVE: Our aim was to quantify thromboxane B2 (TXB2) in umbilical cord serum of term infants of nulliparous, low-risk women who were randomly assigned to either placebo or low-dose (60 mg) aspirin (ASA) on a daily basis from 24 weeks' gestation through delivery as part of a randomized clinical trial for prevention of preeclampsia. METHODS: Umbilical cord sera from 230 singleton, term infants whose mothers were involved in our low-dose ASA trial were assayed for TXB2, the stable metabolite of thromboxane A2, without knowledge of treatment or outcome data. The data were related to assigned treatment group, longitudinal pattern of maternal serum TXB2 levels, and other maternal and newborn characteristics. The data also were analyzed according to whether or not maternal serum levels of TXB2 at 29-31, 34-36, and delivery were reduced > or =50% compared to values prior to initiation of the trial. RESULTS: Umbilical cord TXB2 levels (ng/ml, mean +/- SE) were significantly lower at term in the ASA group (36.1 +/- 3.3, n = 111) than in the placebo group (56.6 +/- 5.7, n = 119; P = 0.002). Umbilical cord TXB2 levels were correlated to those in maternal serum at delivery in the ASA group (r = 0.3441; P = 0.0005) but not in the placebo group (r = 0.0626; P = 0.53). Regardless of assigned treatment group, infants whose mothers had a > or =50% longitudinal reduction in serum TXB2 had lower umbilical cord TXB2 levels (39.2 +/- 3.6, n = 114) than infants whose mothers had <50% reductions in TXB2 (54.6 +/ 5.9, n = 116; P = 0.027). Birthweights of these infants correlated inversely (r = 0.1678, P = 0.017) with maternal serum TXB2 at delivery but not to umbilical cord TXB2 levels; the best correlation between birthweight and maternal serum TXB2 was noted in pregnancies assigned to receive placebo (r = -0.2558, P = 0.009). CONCLUSIONS: Umbilical cord serum levels of TXB2 1) are reduced in instances of long-term maternal ingestion of ASA, 2) correlate well with maternal serum levels of TXB2 at delivery when there is evidence for consistent maternal use of ASA, but 3) do not correlate with maternal serum TXB2 levels when there is no evidence for frequent maternal ingestion of cyclooxygenase inhibitors. These data suggest that the capacity for platelet production of TXA2 in fetal and maternal compartments are regulated independently. Finally, there is an inverse relationship between maternal serum TXB2 levels at delivery and birthweight of newborn infants that is most evident among the pregnancies assigned to placebo and also among pregnancies in which there was little evidence to suggest a pattern of cyclooxygenase inhibitor use during pregnancy. PMID- 11048831 TI - Intrapartum screen for diabetes in patients without prenatal care: use of labor admission serum glucose. AB - OBJECTIVE: For patients presenting in labor with no prenatal care, a rapid screening test for gestational diabetes would potentially aid in decisions for tocolysis (e.g., preterm patients) and mode of delivery (e.g., large for gestational age). We sought to determine whether a labor admission serum glucose is of predictive value in the diagnosis of gestational diabetes. METHODS: We obtained labor admission glucose values for laboring patients and compared these with 1-h (50-g) postglucola (1 degree PG) screens obtained at 24 to 32 weeks' gestation. Diabetics being treated with insulin were excluded from the study. Labor admission serum glucose values were compared to 1 degrees PG values by linear regression. Sensitivity and specificity of admission glucose for identification of a positive 1 degree PG (140 mg/dl) were evaluated by a receiver operator curve (ROC). RESULTS: A total of 98 patients with both 1 degree PG screens and labor admission glucose were identified. Linear regression showed no significant correlation of labor admission glucose and 1 degree PG values (r = 0.13; P = 0.9). The ROC failed to demonstrate an optimal admission random glucose cutoff value for diagnosis of diabetes. CONCLUSIONS: In laboring patients without insulin-requiring diabetes, labor admission glucose does not predict an abnormal 1 degree PG and thus does not aid in labor management of patients with suboptimal prenatal care. PMID- 11048832 TI - Prenatal diagnosis of fetal anomalies in a regional tertiary center: the role of a maternal fetal medicine unit--a review of 6,877 deliveries. AB - OBJECTIVE: To determine the accuracy of anomalies detection and to evaluate the role of maternal-fetal medicine (MFM) specialists. METHODS: This was a retrospective study of birth defects. Patients were divided into: 1) if ultrasound was reviewed by MFM specialists; 2) Others, if reviewed by other ultrasonologists. Fisher's exact test or Pearson's chi2 test were used for statistical analysis. RESULTS: Birth defects occurred in 204/6,877 (3%) neonates with 291 distinct birth defects. Prenatal diagnosis was possible in 181 (62%). In 16 (5.5%) late prenatal diagnosis was possible. MFM specialists correctly diagnosed 53/62 (85%) vs. 56/132 (42%) in Others (P < 0.001). Late diagnosis was possible in 12 cases of gastrointestinal anomalies, three of hydrocephaly and one of skeletal dysplasia; five were correctly diagnosed by MFM specialists and two by Others. CONCLUSIONS: Involvement of ultrasonologists with particular expertise in fetal scanning may improve accuracy of prenatal diagnosis. A repeat third trimester ultrasound may be useful in detecting late-evolving anomalies. PMID- 11048833 TI - Volumetric flow in the umbilical artery: normative data. AB - OBJECTIVE: Provide normative data for the volumetric blood flow (cc/min and cc/min/kg) in the umbilical artery. METHODS: Flow was determined from an umbilical artery in 252 normal obstetrical patients from 18-40 weeks' gestation utilizing pulsed Doppler and color flow Doppler with an angle of insonation of 30 60 degrees. Simultaneous velocimetry studies (S/D ratio, resistance and pulsatility indices), fetal biometry, and an anatomic survey were obtained to further define the normal population. RESULTS: There was a steady increase in the flow (cc/min) in the umbilical artery as pregnancy progressed. Flow/kg showed a steady decline as fetal weight increased. Umbilical artery diameter increased until reaching a plateau at 32-34 weeks. Velocimetric results were consistent with known data. CONCLUSIONS: Volumetric blood flow in the umbilical artery can be determined with relative ease and normative data from 18-40 weeks is presented for the first time. PMID- 11048834 TI - Influence of maternal-fetal medicine subspecialization on the frequency of trial of labor in term pregnancies with breech presentation. AB - OBJECTIVE: To investigate the role of subspecialization in maternal-fetal medicine (MFM) on the frequency of a trial of labor in term pregnancies with breech presentation. METHODS: We conducted a retrospective study of 332 singleton pregnancies > or =37 weeks with nonfootling breech presentation that delivered over a 6-year period (1994-1998) at a university-based, tertiary care hospital. Patients were divided into two groups based on whether the delivery was attended by an MFM or non-MFM obstetrician-gynecologist. Demographic and clinical data were compared between groups and outcome variables included whether the patient had an attempt at vaginal delivery, cesarean delivery after a labor attempt, or vaginal breech delivery. RESULTS: The frequency of labor attempt (OR 1.4, 95% CI 0.9-2.3), vaginal breech success rate (OR 0.6, 95% CI 0.3-1.5), and overall cesarean rates (OR 0.9, 95% CI 0.5-1.7) were similar between groups. Using discriminant function analysis, only nulliparity (R2 = 1.6%, F = 6.0, P = 0.005) and birthweight (R2 = 2.0% F = 6.4, P = 0.01) were associated with trial of vaginal delivery. CONCLUSIONS: Subspecialization in MFM had no impact on the frequency of trial of labor in the term pregnancy with a breech presentation. PMID- 11048835 TI - Two-hour urine collection for evaluating renal function correlates with 24-hour urine collection in pregnant patients. AB - OBJECTIVE: We sought to determine if 2-h urine collection for the assessment of urine protein excretion and creatinine clearance correlates with 24-h urine collection in pregnant patients with renal disease. METHODS: We enrolled patients of gestational ages ranging from 8-41 weeks, admitted as inpatients and having undergone evaluation for renal function (n = 59). We obtained the following samples: 1) 2-h urine, and 2) 24-h urine. We measured serum creatinine concentration, urinary protein, creatinine concentration, and creatinine clearance. The data was analyzed using descriptive analysis, two-way ANOVA, univariate linear regression analysis, and Bland-Altman plot comparing the efficacy of 2-h urine results with 24-h urine results. RESULTS: We analyzed the data on 51 patients. Total proteinuria calculated by protein/creatinine (P:C) ratio in the 2-h group correlated with the total protein measured in the 24-h group (1,840.8 +/- 786 and 1,944 +/- 1,060 mg [mean +/- SE], respectively, r2 = 0.95, P < 0.0001). Creatinine clearance correlated in the 2- and 24-h groups (111 +/- 42 and 122.5 +/- 50 ml/min, respectively; r2 = 0.73, P < 0.001). CONCLUSION: Two-hour urine sampling offers the same clinical information as 24-h urine collection for the evaluation of renal function. Use of 2-h urine collection reduces the time of evaluation and diagnosis, whereby increasing patient compliance and reducing errors in performance of the tests. PMID- 11048836 TI - Obesity-related complications of pregnancy vary by race. AB - OBJECTIVE: To evaluate racial effects on obstetric complications in obese gravidas. METHODS: The obstetric database was reviewed for the period 6/1/94 to 3/31/97. All clinic patients delivering singletons were included. Obesity was defined as a body mass index (BMI) of 29 kg/m2 or more, or a pre-pregnancy weight of 200 pounds or more. Complications studied included hypertension, diabetes, cesarean delivery, and fetal macrosomia. RESULTS: Of 2,424 eligible subjects, 168 were obese (6.9%). Obese patients had higher rates of chronic hypertension and pregestational diabetes, as well as increased rates of preeclampsia, gestational diabetes, fetal macrosomia, cesarean delivery, and operative vaginal delivery compared to nonobese patients. Of the obese patients, 105 (63%) were Hispanic, 39 (23%) were African American, and 24 (14%) were White; no Asian or Mixed/Other patients were obese. Mean BMIs of the obese subgroups did not differ (P = 0.14), but prepregnancy weights were greater in Whites than Hispanics (P < 0.002). Obese Hispanics had an increased rate of gestational diabetes (P = 0.04) and of infant weight > or =4,500 g (P =.03). Obese Hispanic and African American women were more likely than obese Whites to deliver by cesarean (P = 0.03). CONCLUSION: Racial differences affect the complication rates in obese gravidas, and may influence prenatal counseling and pregnancy management. PMID- 11048837 TI - Central nervous system complications of von Hippel-Lindau disease and pregnancy; perinatal considerations: case report and literature review. AB - A 30-year-old woman with von Hippel-Lindau disease presented at 30 weeks' gestation with a symptomatic cerebellar hemangioblastoma. She underwent a craniotomy for complete removal of the tumor. The postoperative period and the remaining of the pregnancy were uneventful. She delivered under epidural anesthesia after induction for postterm. PMID- 11048838 TI - Recurrent acquired sideroblastic anemia in a twin pregnancy. AB - A woman whose sideroblastic anemia had relapsed with progestogen and combined oral contraceptive therapy suffered further relapses in a (twin) pregnancy. Previous reports exist of relapses both from progestogens and in pregnancy, and we postulate a shared etiology. Affected women considering pregnancy or sex hormone usage should be advised accordingly. PMID- 11048839 TI - Three-week delayed delivery in a triplet pregnancy at full dilatation with aggressive tocolysis. AB - We describe our aggressive management of a woman with a triplet pregnancy resulting from in vitro fertilization who presented at 22 weeks' gestation in labor with full cervical dilation. Successful multiagent tocolysis and delayed delivery of the two remaining fetuses after delivery of the presenting fetus led to an improved outcome for all three babies. PMID- 11048840 TI - Methylene blue-induced hyperbilirubinemia in neonatal glucose-6-phosphate dehydrogenase (G6PD) deficiency. AB - Methylene blue continues to be used in the gravid female. We report three premature neonates exposed to methylene blue that experienced severe hemolytic reactions requiring exchange transfusions. Two neonates were subsequently diagnosed with G6PD deficiency. Continued caution is warranted prior to the use of methylene blue in the gravid female. PMID- 11048841 TI - Liquid chromatographic determination of nicarbazin in feeds. AB - A new liquid chromatographic method has been developed for determination of nicarbazin in feeds. Approximately 40 g feed is extracted with 200 mL acetonitrile-water (80 + 20, v/v). An aliquot of the extract is filtered and assayed using a reversed-phase isocratic method that measures the 4,4' dinitrocarbanilide moiety of nicarbazin at a wavelength of 340 nm. For medicated feeds, the method uses a standard linear range of 5 to 100 microg/mL. For lower levels, a linear range of 50 to 150 ng/mL can be used. The method has a limit of detection of 250 ng/g and a limit of quantitation of 500 ng/g in a 40 g feed sample. Recovery was 99.1%, with a range of 95.2 to 101.8%. In the typical U.S. dosing range of 27 to 113.5 g/ton, the precision of the method based on one analyst, one day, and 2 weighings ranged from 2.8% (113.5 g/ton) to 4.7% (27 g/ton). PMID- 11048842 TI - Improved diffusion methods for nitrogen and 15nitrogen analysis of Kjeldahl digests. AB - Simple methods are described that permit the use of either H3BO3 indicator solution or acidified filter disks to collect NH3 liberated by treatment of Kjeldahl digests with NaOH. These methods incorporate modifications to improve reliability, analytical capacity, and convenience. A semimicro digest was diluted to 25 mL with deionized water, and a 10 mL aliquot, containing up to 4 mg N (150 microg N for diffusions into acidified disks), was transferred to a shell vial, which was placed inside a 473 mL (1 pint) Mason jar containing 10 mL 10N NaOH. The NH3 liberated by overturning the vial was collected after 12 to 48 h at ambient temperature, or after 4 h at 45 to 50 degrees C on a hotplate, for quantitative and/or isotope-ratio analyses. With either H3BO3 indicator solution or acidified filter disks, recovery of diffused N was quantitative. Isotope-ratio analyses of diffused N from 15N-labeled chemical, plant, and soil samples were within 3% of analyses using steam distillation. PMID- 11048843 TI - Simultaneous determination of chlorothalonil and hexachlorobenzene in technical and formulated materials by capillary gas chromatography: collaborative study. AB - A collaborative study was conducted for the capillary gas chromatographic (GC) method for the simultaneous determination of the fungicide chlorothalonil (CTL) and the accompanying impurity, hexachlorobenzene (HCB), in technical and formulated materials. The method calls for the dissolution of technical and dry formulations of CTL and HCB from the aqueous flowable formulation. The 10 participating laboratories were asked to analyze the samples by adhering to the method as closely as their instrumentation and data systems allowed, and to note any deviations from the method. Collaborators were asked to prepare the standards and samples, set up the capillary GC systems, analyze the samples, and calculate the results. CTL produced reproducibility relative standard deviations (RSDR) of 0.4-2.5 (active ingredient concentrations ranged from approximately 52 to 98% by weight). HCB produced RSDR values of 5.2-22% (HCB concentrations were 0.02-0.04% by weight). The method was adopted First Action by AOAC INTERNATIONAL. PMID- 11048844 TI - Quantitative in vitro bioassay for recombinant human interleukin-11. AB - A cell culture-based in vitro bioassay was developed to measure the biological activity of recombinant human interleukin-11 (rhIL-11). The bioassay measures induced proliferation of T10 cells, derived from the T1165 murine plasmacytoma line. A colorimetrically detectable formazan product, obtained by cellular reduction of the tetrazolium compound, 4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5 tetrazolio]-1,3-benzene disulfonate (WST-1), was used as an endpoint for response of clone T10 to added rhIL-11. Positions of the samples and the standards in 96 well microplates affected the precision of this bioassay, which was improved by using 2 microplates where serially diluted sample and standard lines were interleaved and their positions were alternated. The coefficient of variation for this bioassay was less than 8%. This method is suitable for quality control of rhIL-11 because of its simplicity, reproducibility, and accuracy. PMID- 11048845 TI - Environmental screening of acidic compounds based on capillary zone electrophoresis/laser-induced fluorescence detection with identification by gas chromatography/mass spectrometry and gas chromatography/high-resolution mass spectrometry. AB - This paper describes the application of capillary zone electrophoresis/laser induced fluorescence detection (CZE/LIF) to the discovery of acidic compounds in environmental matrixes or the screening of extracts for acidic components. Published studies indicate that coal-derived materials contain a significant fraction of acidic compounds relative to materials derived from petroleum and shales. Such compounds may be useful as marker compounds for site assessment and source apportionment issues, and their identification may be important in toxicological and other health issues. We used deep-UV light from the frequency doubled Ar ion laser at 244 and 257 nm to study extracts of samples. The CZE/LIF technique possesses good sensitivity and therefore overcomes one of the limitations of CZE with UV detection. The present work depends on high pressure/temperature solvent extraction of polynuclear aromatic hydrocarbon (PNA) contaminated soil, followed by separation using CZE. The anionic analytes were separated by using borate or phosphate buffer (pH 9.2-12.3) after a chemical class separation. Samples were also characterized by gas chromatography/mass spectrometry (GC/MS) using full scans at low resolution, and elemental compositions were determined unequivocally by GC/high-resolution MS (GC/HRMS) using mass peak profiling (MPP). The similarity of low-resolution electron ionization mass spectra for a standard, 1-hydroxypyrene, and for a series of compounds in a contaminated-soil extract suggested that several types of phenolic and hydroxy-PNAs were present, including hydroxylated derivatives of fluorenes, fluoranthenes, and pyrenes. GC/HRMS using MPP confirmed the elemental compositions of the hydroxyfluorenes and hydroxypyrenes (and presumably hydroxyfluoranthenes) as [C13H10O] and [C16H10O], respectively. A new version of the MPP software was written for the Finnigan-MAT 900S-Trap and was similar to that developed previously for the VG 250SE. Inclusion of a calibration ion in addition to a lock mass ion in the multiple-ion detection descriptor provided errors of <1 ppm for the 3 partial profiles of the analytes. A mass resolution of 31,000 was used to resolve the analyte signals from interferences evident in the full M+1 and M+2 profiles in the case of the hydroxyfluorenes. Derivatization was also performed to form the tert-butyldimethylsilyl derivatives of phenolic hydroxy groups as a further confirmation of structure. PMID- 11048846 TI - Determination of binary and ternary mixtures of pesticides in wetland waters by gas chromatography using partial least-squares analysis. AB - Partial least-squares (PLS) analysis was applied for the first time to ternary and binary mixtures of pesticides producing overlapping peaks in gas chromatograms. Several preprocessing algorithms for pretreatment of data were tested for optimization of the PLS models, and the most advantageous were used in each case. Three different mixtures of pesticides were resolved with satisfactory results: parathion-methyl, chlorpyrifos-methyl, and vinclozolin; parathion-ethyl, chlorpyrifos, and triadimefon; and endosulfan sulfate and carbophenothion. The proposed models were applied to the determination of 3 mixtures of pesticides at levels of 0.02-1.00 ng/mL in wetland water samples after a preconcentration step with C18 cartridges, with recoveries ranging from 82.7 to 117.4%. The reported method is sufficiently sensitive to measure pesticide residues at the maximum allowable concentrations given in the European Union Drinking Water Directive. PMID- 11048848 TI - Determination of Se4+ in drinkable water by solid-phase microextraction and gas chromatography/mass spectrometry. AB - A method was developed for the selective determination of Se4+ in drinkable water by solid-phase microextraction (SPME) and gas chromatography/mass spectrometry (GC/MS). Se4+ was selectively derivatized to ethane, 1,1'-selenobis by reaction with sodium tetraethylborate, extracted by the SPME fiber, and determined by GC/MS. Both headspace (HS)-SPME and direct SPME were studied. The method requires only a few milliliters of sample and 20 min for completion. At 2.0 microg/L concentration, the relative standard deviation was 10.1% for HS-SPME and 9.1% for direct SPME. For HS-SPME, the theoretical detection limit was 81 ng/L and 166 ng/L for direct SPME. The recovery rate was 95%. The method was used to determine Se4+ in 10 tap water samples. PMID- 11048847 TI - Liquid chromatographic determination of triasulfuron in soil. AB - Two extraction methods were developed for the determination of triasulfuron in soil. Method I included extraction with methanol-phosphate buffer at pH 7 (2 + 1, v/v), liquid-liquid partition with dichloromethane, and cleanup on a liquid chromatographic Si adsorption solid-phase extraction tube. In Method II, Extrelut was added and the sample was then extracted with acetonitrile. In both cases, the extracts were analyzed by liquid chromatography (LC) with UV detection and the LC peak was confirmed by LC/mass spectrometry (MS). The 2 methods were tested on 3 soils having different physicochemical characteristics. Method I gave 83% average recovery and a determination limit of 0.4 microg/kg soil. Method II gave 67% average recovery and a determination limit of 2 microg/kg soil. Examples of application of Method I to field samples are reported. PMID- 11048849 TI - Minimum detectable level of Salmonellae using a binomial-based bacterial ice nucleation detection assay (BIND). AB - A modified bacterial ice nucleation detection (BIND) assay was used for rapid and sensitive detection of several Salmonella species. For the BIND assay, Salmonella cells are infected with bacteriophage genetically modified to contain DNA encoding an ice nucleation protein (INP). After infection, de novo protein synthesis occurs and INPs are incorporated into the outer membrane of the organism. After supercooling (-9.3 degrees C), only buffer solutions containing transfected salmonellae freeze, causing a phase-sensitive dye to change color. This technique, and a probability-based protocol modification, provided quantitative detection with a minimum detectable level (MDL) of 2.0 +/- 0.3 S. enteritidis cells/mL in buffer (about 3 h). The MDLs for S. typhimurium DT104 and S. abaetetuba were 4.2 +/- 0.2 and 11.1 +/- 0.4 cells/mL, respectively. Using salmonellae-specific immunomagnetic bead separation technology in conjunction with the modified BIND protocol, we achieved an MDL of about 4.5 S. enteritidis cells/mL with an apparent capture efficiency of 56%. PMID- 11048850 TI - Evaluation of a dry, rehydratable film method for rapid enumeration of Staphylococcus aureus. AB - Results with the new 3M Petrifilm Rapid S. aureus Count (RSA) Plate method were compared with those of the classical Baird-Parker agar (BPA) method for detection and enumeration of Staphylococcus aureus. Studies on 219 bacterial strains demonstrated that the Petrifilm RSA plate is more sensitive than and as specific as the classical BPA method for confirmed identification of S. aureus. Counts of colonies from 71 pure cultures, 61 naturally contaminated food samples, and more than 750 artificially inoculated food samples showed that the Petrifilm RSA method was as effective as the classical BPA method for identification and enumeration of S. aureus. The Petrifilm RSA method gave results in one-third the time required for the classical method. PMID- 11048851 TI - Enumeration of Cryptosporidium spp. in water with US EPA method 1622, USA. AB - The occurrence of Cryptosporidium parvum or other pathogenic Cryptosporidium species in water must be known in order to assess risk and determine the treatment needed to reduce Cryptosporidium oocysts to acceptable levels in finished drinking water. Because Cryptosporidium oocyst occurrence may be sparse, methods must concentrate a large volume of water and correctly identify oocysts in the concentrate. The U.S. Environmental Protection Agency Information Collection Rule (ICR) protozoan method gives low and variable recoveries of Cryptosporidium oocysts, making risk assessment difficult. Therefore, a method giving better oocyst recovery and more consistent results was needed. Method 1622 was developed with existing materials and procedures, and improvements were made in filtration, cleanup, and detection. Absolute porosity filters were used, with cleanup by immunomagnetic separation and detection by direct fluorescent antibody stain with 4',6-diamidino-2-phenylindole (DAPI) staining for additional cell structures. Both the level and consistency of oocyst recovery were improved compared to recovery with the ICR method. PMID- 11048852 TI - Isolation of Alicyclobacillus acidoterrestris from fruit juices. AB - K agar, a novel isolation medium developed for the food industry, was compared with other acidified media for isolation of Alicyclobacillus acidoterrestris spores. Spores were inoculated into apple juice, orange juice, and a fruit juice blend and then isolated on the following media: K agar, pH 3.7; semi-synthetic medium, pH 4.0; orange serum agar, pH 3.5; and minimal salts medium, pH 4.0. Media were incubated at 24, 35, 43, and 55 degrees C. Highest recovery of spores was obtained with either K agar or semi-synthetic medium, incubated at 43 degrees C. The effect of heat shocking spores at different times was also determined; heat shocking at 80 degrees C for 10 min was considered appropriate. Peptone, previously shown to inhibit A. acidoterrestris, was not inhibitory when present in K agar. A collaborative trial with 9 laboratories was undertaken to determine the repeatability and reproducibility of counts on K agar. K agar prepared from individual components was compared with dehydrated K agar prepared by International BioProducts (Redmond, WA). There were no significant differences between log mean counts for the 2 media for each of the juices analyzed at both the high and the low inoculum levels. Repeatability and reproducibility values were not significantly different either within juices, within trials, or across all samples tested in both trials. K agar is suitable for isolation of A. acidoterrestris spores from fruit juices. PMID- 11048854 TI - Hydrolysis and redox factors affecting analysis of common phenolic marker compounds in botanical extracts and finished products. AB - Many of the marker compounds analyzed in herbal products are redox-active phenolic molecules, which are commonly found in plants as components of glycosides and starch polymers. Variability in degree of sample hydrolysis can occur due to differences in water content, pH, and temperature. Sonication versus shaking during extraction can also influence hydrolysis and oxidation of sensitive compounds. Some traditional botanical extract marker compounds are esters and glycosides of phenolics such as echinacoside from Echinacea while others are free phenolics, such as quercetin from glycosides in Ginkgo. Optimizing hydrolysis conditions maximizes free quercetin levels, but lowers echinacoside levels. Furthermore, acidic hydrolysis conditions mimic stomach conditions encountered by oral supplements and protect resulting free phenolics from oxidation. Oxidative degradation of botanical phenolic markers can be initiated by light, sonication, oxygen, basic pH conditions, heat, redox-active solvents, and formulation additives. Some phenolic markers reversibly cycle through multiple oxidation states creating a formula-specific equilibrium of oxidation states. Finished product formulations that include easily oxidized phenolics, carbonates, phosphates, and transition metals affect sample hydrolysis degree and redox equilibria, and quantitation. By recognizing and controlling hydrolysis and oxidation variables, more accurate and rugged methods can be developed allowing for improved botanical standardization and finished product analysis. PMID- 11048853 TI - Revalidation and long-term stability of National Institute of Standards and Technology Standard Reference Materials 1566, 1567, 1568, and 1570. AB - Multiple units of Standard Reference Materials (SRMs) 1566 Oyster Tissue, 1567 Wheat Flour, 1568 Rice Flour, and 1570 Trace Elements in Spinach, produced by the National Institute of Standards and Technology (NIST, then the National Bureau of Standards), were analyzed 17-20 years after the original certification dates and 12-15 years after the certificates became invalid. Instrumental neutron activation analysis and thermal neutron prompt gamma-ray activation analysis were used to measure mass fractions for 27 elements in these SRMs to revalidate them for use in quality assurance (QA) programs required for food analysis programs within the U.S. Food and Drug Administration. With the exception of Se in SRM 1567, all element mass fractions were in agreement with certified values and literature data. Some evidence of B loss from SRM 1568 was observed. These materials were judged to be suitable for continued use in QA programs. Findings showed that these matrixes exhibited stability of moisture, mass fraction, and weight basis for far longer (> or =15 years) than was indicated by the 5-year validity statement on the NIST Certificates of Analysis. PMID- 11048855 TI - Determination of biotin and folate in infant formula and milk by optical biosensor-based immunoassay. AB - Biomolecular interaction analysis was evaluated for the automated analysis of biotin- and folate-supplemented infant formulas and milk powders. The technique was configured as a biosensor-based, nonlabeled inhibition immunoassay using monoclonal antibodies raised against analyte-conjugate. Sample extraction conditions were optimized and antibodies were evaluated for cross-reactivity. Performance parameters included a quantitation range of 2-70 ng/mL, recoveries of 86-102%, agreement against assigned reference values for National Institute of Standards and Technology Standard Reference Material 1846, between-laboratory reproducibility relative standard deviation of 9.1% for biotin and 8.1% for folate, respectively, and equivalence against reference microbiological assay methods for both analytes. PMID- 11048856 TI - Determination of sulfite in Oriental herbal medicines. AB - Sulfite was detected in 7 varieties of Oriental herbal medicines (Pueraria radix, Zingiberis rhizoma, Platycodon radix, Adenophora radix, Pinellia tuber, Astragalus radix, and Paeonia radix) on the Korean market. Sulfiting of commercial Oriental herbal medicines by fumigation with burning bituminous coal was simulated, and the accumulation of sulfite was investigated by using fresh Platycodon radix roots obtained from a growing field. The sulfite level reached a plateau in 9 h, and the maximum sulfite level found by the Monier-Williams (MW) method (AOAC 990.28) was 1020 ppm. The sulfite content in the simulated Platycodon radix sample determined by alkali extraction followed by ion-exclusion chromatography with electrochemical detection (AOAC 990.31) was approximately 17% lower on average than the MW results. Free-sulfite levels determined by acid extraction and ion-exclusion chromatography with electrochemical detection were between 19 and 49% of the MW results. The advantages of different methods for sulfite determination and the significance of the results are discussed. PMID- 11048857 TI - Determination of naringin and neohesperidin in orange juice by liquid chromatography with UV detection to detect the presence of grapefruit juice: Collaborative Study. AB - Fifteen collaborating laboratories were sent 9 samples of citrus juice mixtures as blind duplicates for determination of naringin and neohesperidin by liquid chromatography. Two sample pairs were 100% orange juice and did not contain any naringin or neohesperidin. The remaining 7 sample pairs contained naringin at levels ranging from 3.9 to 46.5 ppm and neohesperidin at levels ranging from 0.14 to 35.6 ppm. Five sample pairs consisted of orange juice mixtures containing 1, 3, and 5% grapefruit juice; 5% sour orange; and 5% K-Early citrus variety. Two sample pairs were orange juice spiked with naringin, neohesperidin, sodium benzoate, and potassium sorbate. Data were received from 13 laboratories. Data from 1 collaborator were eliminated because the method protocol was not followed. Neohesperidin values from another laboratory were also not used because of problems with a coeluting component. Repeatability relative standard deviations ranged from 2.95 to 15.23% for naringin and from 3.00 to 11.74% for neohesperidin. Reproducibility relative standard deviations ranged from 11.34 to 31.94% for naringin and from 10.45 to 26.17% for neohesperidin. The method is reliable for detecting the presence of grapefruit juice in orange juice as indicated by a finding of > or =10 ppm naringin and < or =2 ppm neohesperidin. The method was adopted First Action by AOAC INTERNATIONAL. PMID- 11048858 TI - Determination of inorganic cations by capillary ion electrophoresis in Ilex paraguariensis (St. H.), a plant used to prepare tea in South America. AB - A practical and economical capillary ion electrophoresis method with indirect UV detection at 214 nm was developed for determination of inorganic cations in plants of Ilex paraguariensis(St. H.) and their infusion known as mate tea, a very popular beverage in South America. A microwave digestion procedure was used to prepare the herbal plants, but the infusion was only diluted. The background electrolyte contained 6mM imidazole and 10mM alpha-hydroxyisobutyric acid, pH 4.0. The running voltage was 20 kV and temperature was 25 degrees C. K, Na, Ca, Mg, and Mn ions were quantitated, and linearity was demonstrated between 0.6 and 120 ppm. The results were in good agreement with those obtained by flame atomic absorption and emission spectrometry. Accuracy of the method was verified by comparison with Beech leaves CRM 100, a standard reference material. The high content of minerals and several oligoelements, especially Mn in mate tea, is considered to be of nutritional interest. PMID- 11048859 TI - Graphite furnace atomic absorption spectrometric determination of lead and cadmium extracted from ceramic foodware: Collaborative Study. AB - A modification of the official flame atomic absorption spectrometric (FAAS) method for determining lead and cadmium extracted from ceramic foodware was collaboratively studied. In the modified method, graphite furnace atomic absorption spectrometry (GFAAS) is substituted for FAAS. The modified method also includes mandatory quality control procedures to improve method performance. The extraction procedure of the official method (leaching with 4% acetic acid for 24 h at room temperature) remains unchanged. Seven laboratories analyzed blind duplicate portions of 3 ceramicware leach solutions containing Pb at concentrations of 0.0196, 0.403, and 3.73 microg/mL and Cd at concentrations of 0.00236, 0.0456, and 0.544 microg/mL. Performance of the modified method compared well with that of the official method. The repeatability relative standard deviation (RSDr) ranged from 0.87 to 6.7% for Pb and from 3.7 to 11% for Cd. The reproducibility relative standard deviation (RSDR) ranged from 4.5 to 12% for Pb and from 7.0 to 11% for Cd. Accuracy of collaborator results was 97-98% for Pb and 93-101% for Cd. Quality control results and quantitation limits were excellent. Method quantitation limits varied among laboratories from 0.005 to 0.019 microg/mL for Pb and from 0.0004 to 0.0019 microg/mL for Cd. The modified method was adopted First Action by AOAC INTERNATIONAL. PMID- 11048860 TI - Determination of lead, cadmium, zinc, copper, and iron in foods by atomic absorption spectrometry after microwave digestion: NMKL Collaborative Study. AB - A method for determination of lead, cadmium, zinc, copper, and iron by atomic absorption spectrometry (AAS) after microwave digestion was subjected to a collaborative study in which 16 laboratories participated [including users of inductively coupled plasma (ICP) and ICP-mass spectrometry (MS)]. The types of samples included in the study were minced fish, wheat bran, milk powder, bovine and pig liver, mushroom, 2 simulated diets, and bovine muscle; the last 4 were certified reference materials. These were analyzed as single (4 samples), double blind (1 sample), or split level (2 samples) samples. Before the collaborative study, a pretrial was conducted in which 4 ready-made solutions and one fish tissue sample were analyzed for Pb and Cu. The reproducibility relative standard deviation (RSDR) values, for results above the detection limit, ranged from 59% at 0.155 mg/kg to 16% at 1.62 mg/kg for Pb, from 28% at 0.0124 mg/kg to 11% at 0.482 mg/kg for Cd, from 9.3% at 35.3 mg/kg to 1.7% at 147 mg/kg for Zn, from 39% at 0.241 mg/kg to 3.0% at 63.4 mg/kg for Cu, and from 17% at 7.4 mg/kg to 5.9% at 303 mg/kg for Fe. The RSDR values agreed well with the norms described by the International Union of Pure and Applied Chemistry. As a complement to the AAS determinations, a number of laboratories analyzed the samples either by ICP or by ICP-MS. The results of these analyses agreed well with the AAS results. On the basis of the results of the collaborative study, the method was adopted Official First Action by AOAC INTERNATIONAL. PMID- 11048861 TI - Determination of metals in foods by atomic absorption spectrometry after dry ashing: NMKL Collaborative Study. AB - A method for determination of lead, cadmium, zinc, copper, and iron in foods by atomic absorption spectrometry (AAS) after dry ashing at 450 degrees C was collaboratively studied in 16 laboratories. The study was preceded by a practice round of familiarization samples and another round in which solutions were distributed and the metals were determined directly by AAS. The study included 5 different foods (liver paste, apple sauce, minced fish, wheat bran, and milk powder) and 2 simulated diets. A single analysis was carried out with each sample. Suitable sample combinations were used as split-level combinations for determination of the repeatability standard deviation. The reproducibility relative standard deviation for each of the elements ranged from 20 to 50% for lead concentrations of 0.040-0.25 mg/kg, from 12 to 352% for cadmium concentrations of 0.001-0.51 mg/kg, from 4 to 8% for zinc concentrations of 0.7 38 mg/kg, from 7 to 45% for copper concentrations of 0.51-45 mg/kg, and from 11 to 14% for iron concentrations of 4-216 mg/kg. PMID- 11048862 TI - Determination of oxalate in urine, using an amperometric biosensor with oxalate oxidase immobilized on the surface of a chromium hexacyanoferrate-modified graphite electrode. AB - A novel enzymatic amperometric method is described for the determination of oxalic acid in urine. An amperometric biosensor was made by immobilizing oxalate oxidase on the surface of a chromium(III) hexacyanoferrate-modified graphite electrode by using a bovine serum albumin and glutaraldehyde cross-linking procedure. The enzyme biocatalyzes oxalate decomposition in the presence of oxygen into carbon dioxide and hydrogen peroxide. The oxalate concentration, which is proportional to the amount of hydrogen peroxide, was determined amperometrically by measuring the current resulting in the reduction of hydrogen peroxide at a very low working potential (0.05 V versus the Hg ?Hg2Cl2? 3M KCl electrode), which minimized the influence of the possible interferences present in human urine. All experiments were performed with succinic buffer, pH 3.8, containing 0.1M KCl and 5.4mM ethylenediaminetetraacetic acid. In an aqueous solution of pure oxalic acid, the biosensor showed good linearity in a concentration range of 2.5-100 microM without the use of a dialysis membrane. For untreated urine samples, a high correlation (R2 = 0.9949) was obtained between oxalate concentrations added to urine samples and oxalate recoveries calculated for determinations with the described oxalate biosensor. PMID- 11048863 TI - Determination of thirteen common elements in food samples by inductively coupled plasma atomic emission spectrometry: comparison of five digestion methods. AB - Five sample digestion procedures were evaluated for the determination of Al, B, Ca, Cu, Fe, K, Mg, Mn, Na, P, S, Sr, and Zn in food samples by inductively coupled plasma atomic emission spectrometry. The 5 procedures include dry ashing at 500 degrees C, wet digestion with HNO3-HClO4, microwave digestion with HNO3, microwave digestion with HNO3-H2O2, and microwave digestion with HNO3-H2O2-HF. For microwave digestion with HNO3-H2O2-HF, silicon (IV) oxide was used to eliminate the excess HF, making it possible to determine total Al, B, and other common elements accurately and simultaneously. Seven National Institute of Standards and Technology standard reference materials (SRMs) were analyzed to compare the recovery of 13 elements with above digestion procedures. The results demonstrated that the microwave digestion procedure with HNO3-H2O2-HF yielded the best recoveries for all 13 elements in the selected SRMs. The determined concentrations of most elements were close for all 3 microwave digestion procedures with the exception of Al in oyster tissue, bovine liver, and spinach. Notably, the wet digestion with HNO3-HClO4 is the simplest and the most effective procedure for the selected elements except Al and B. Although there are several concerns with the dry ashing procedure, it might be a preferable procedure for those analyses where only nonvolatile elements are to be determined and the concentrations of the elements are low. PMID- 11048864 TI - Determination of fluvalinate residues in beeswax by gas chromatography with electron-capture detection. AB - A simple, rapid, and accurate method is described for the determination of residual fluvalinate in beeswax. The procedure consists of partitioning on a disposable column of diatomaceous earth (Extrelut), followed by chromatographic cleanup on a Florisil cartridge. The final extract is analyzed by capillary gas chromatography with electron-capture detection (GC-ECD). Briefly, wax samples were dissolved in n-hexane, and the solutions were sonicated and transferred to Extrelut columns. The fluvalinate was extracted with acetonitrile, and a portion of the extract was cleaned up on a Florisil cartridge. The fluvalinate was eluted with diethyl ether-n-hexane (1 + 1) and directly determined by GC-ECD. Recoveries from wax samples spiked at 5 fortification levels (100-1500 microg/kg) ranged from 77.4 to 87.3%, with coefficients of variation of 5.12-8.31%. The overall recovery of the method was 81.4 +/- 3.2%, and the limit of determination was 100 microg/kg. PMID- 11048865 TI - Determination of two oxy-pyrimidine metabolites of diazinon in urine by gas chromatography/mass selective detection and liquid chromatography/electrospray ionization/mass spectrometry/mass spectrometry. AB - An analytical method was developed for the determination in urine of 2 metabolites of diazinon: 6-methyl-2-(1-methylethyl)-4(1H)-pyrimidinone (G-27550) and 2-(1-hydroxy-1-methylethyl)-6-methyl-4(1H)-pyrimidinone (GS-31144). Two of the urine sample preparation procedures presented rely on gas chromatography/mass selective detection (GC/MSD) in the selected ion monitoring mode for determination of G-27550. For fast sample preparation and a limit of quantitation (LOQ) of 1.0 ppb, urine samples were purified by using ENV+ solid-phase extraction (SPE) columns. For analyte confirmation at an LOQ of 0.50 ppb, classical liquid/liquid partitioning was used before further purification in a silica SPE column. An SPE sample preparation procedure and liquid chromatography/electrospray ionization/mass spectrometry/mass spectrometry (LC/ESI/MS/MS) were used for both G-27550 and GS-31144. The limit of detection was 0.01 ng for G-27550 with GC/MSD, and 0.016 ng when LC/ESI/MS/MS was used for both G-27550 and GS-31144. The LOQ was 0.50 ppb for G-27550 when GC/MSD and the partitioning/SPE sample preparation procedure were used, and 1.0 ppb for the SPE only sample preparation procedure. The LOQ was 1.0 ppb for both analytes when LC/ESI/MS/MS was used. PMID- 11048866 TI - Determination of simazine, terbuthylazine, and their dealkylated chlorotriazine metabolites in soil using sonication microextraction and gas chromatography. AB - A rapid analytical method is proposed for the determination of simazine, terbuthylazine, and their chloro dealkylated metabolites (simazine-desethyl, simazine-bisdesethyl, and terbuthylazine-desethyl) in soil. A sonication micromethod is presented for the extraction of -triazine herbicides and their metabolites. Final determination is by gas chromatography (GC) with nitrogen phosphorus detection. The identity of all compounds was confirmed by GC with mass selective detection in the selected-ion monitoring mode. All chromatograms were very clean, without interfering peaks, and no cleanup was needed. The limits of detection were 1 pg for simazine-bisdesethyl; 5 pg for simazine, terbuthylazine, and terbuthylazine-desethyl; and 10 pg for simazine-desethyl. The limits of quantitation were 1, 5, and 10 ppb, respectively. Mean recoveries from fortified soils ranged from 76% for simazine-bisdesethyl to 102% for simazine-desethyl, with relative standard deviations of 3-6%. PMID- 11048867 TI - Sampling agricultural commodities for mycotoxins. PMID- 11048868 TI - Performance of three pneumatic probe samplers and four analytical methods used to estimate aflatoxins in bulk cottonseed. AB - The requirement by the U.S. Food and Drug Administration that agricultural products susceptible to aflatoxin contamination contain aflatoxin at levels < or =20 parts per billion for consumer-ready products has led to the establishment of inspection programs by various industries. In Arizona, cottonseed samples from 100 ton piles are collected by an accumulation of 3 or more probings with a pneumatic probe. When sampling compacted cottonseed piles, the large official pneumatic probe (7.6 x 127 cm) decreases in efficiency. Two smaller probes (1.9 x 127 cm and 1.9 x 254 cm ) were therefore developed and tested for their suitability for sampling cottonseed piles. Three rapid analytical methods (one thin-layer chromatographic and 2 immunochemical) were tested for suitability as on-site assay systems. An analysis of variance of the analytical test results showed no differences between the various probes tested. Of the rapid methods, however, only the AflaTest-P immunoaffinity column gave results similar to those of the official AOAC thin-layer chromatography method. In terms of safety, however, all methods prevent material contaminated above regulatory limits from reaching the consumer. PMID- 11048869 TI - Sampling plans for the determination of aflatoxin B1 in large shipments of animal feedstuffs. AB - Incremental samples (50, 100, and 500 g) were systematically collected from large shipments of copra meal pellets, copra cake, and palm kernel cake to study the distribution of aflatoxin B1 and evaluate adherence of distribution to the model, CV(2)is (EQ) = A + B/Mis (where CVis = coefficient of variation of the true concentration of aflatoxin B1 within the incremental samples; Mis = mass of the incremental samples; and A and B are constants). Also evaluated was the distribution of aflatoxin B1 among 1 kg composite samples, produced both by random combination of existing incremental samples and by collection of 1 kg composite samples (composed of 10 x 100 g increments) from additional batches of copra meal pellets and cottonseed cake. The efficiency of selected sample preparation (grinding and subdivision) procedures was compared, culminating in the development and description of a variety of sampling plans. The coefficient of variation (CV) among incremental samples varied from 0 to 38%, and was independent of incremental sample size. No significant difference (F-test, 5% significance level) was found between the efficacy of 4 sample preparation methods when these methods were applied to the commodities described above. Various sampling plans were evaluated with estimated CVs from 4.0 to 12.5%, for the aflatoxin B1 content of the composite samples. PMID- 11048870 TI - Variability of deoxynivalenol measurements in barley. AB - Deoxynivalenol (DON), a toxin produced by Fusarium fungi, can occur in many cereal grains. If wet climatic conditions coincide with the flowering period of plant development, circumstances are favorable for the fungi infection. Because the presence of DON in barley can have significant economic consequences to barley producers, commercially available test kits are used to measure DON in shipments throughout marketing channels. The quantity of barley sampled from a lot and used to measure DON can vary widely, depending on where the test is conducted. The Grain Inspection, Packers and Stockyards Administration of the U.S. Department of Agriculture specifies that a minimum of 100 g of grain must be processed to measure DON. Other laboratories may use more or less grain. A study undertaken to measure the variability among measurements of different sample sizes found no detectable differences in variability attributable to sample size. It was concluded that the variability among DON concentrations in samples from the lot was small relative to that introduced by the measurement process (combined sample preparation and analysis). A separate experiment investigated variation among samples taken from a lot, variation among subsamples taken from ground samples, and variation among multiple replicated measurements of an extract. On 10 lots, all 3 sources were significant contributors to variation. Stratification of DON within lots was hypothesized as a source of variation of DON measurements. Tests indicated that some stratification may exist. PMID- 11048871 TI - Testing shelled corn for aflatoxin, Part I: estimation of variance components. AB - The variability associated with testing lots of shelled corn for aflatoxin was investigated. Eighteen lots of shelled corn were tested for aflatoxin contamination. The total variance associated with testing shelled corn was estimated and partitioned into sampling, sample preparation, and analytical variances. All variances increased as aflatoxin concentration increased. With the use of regression analysis, mathematical expressions were developed to model the relationship between aflatoxin concentration and the total, sampling, sample preparation, and analytical variances. The expressions for these relationships were used to estimate the variance for any sample size, subsample size, and number of analyses for a specific aflatoxin concentration. Test results on a lot with 20 parts per billion aflatoxin using a 1.13 kg sample, a Romer mill, 50 g subsamples, and liquid chromatographic analysis showed that the total, sampling, sample preparation, and analytical variances were 274.9 (CV = 82.9%), 214.0 (CV = 73.1 %), 56.3 (CV = 37.5%), and 4.6 (CV = 10.7%), respectively. The percentage of the total variance for sampling, sample preparation, and analytical was 77.8, 20.5, and 1.7, respectively. PMID- 11048872 TI - Testing shelled corn for aflatoxin, Part II: modeling the observed distribution of aflatoxin test results. AB - The suitability of several theoretical distributions to predict the observed distribution of aflatoxin test results in shelled corn was investigated. Fifteen positively skewed theoretical distributions were each fitted to 18 empirical distributions of aflatoxin test results for shelled corn. The compound gamma distribution was selected to model aflatoxin test results for shelled corn. The method of moments technique was chosen to estimate the parameters of the compound gamma distribution. Mathematical expressions were developed to calculate the parameters of the compound gamma distribution for any lot aflatoxin concentration and test procedure. Observed acceptance probabilities were compared to operating characteristic curves predicted from the compound gamma distribution, and all 18 observed acceptance probabilities were found to lie within a 95% confidence band. The parameters of compound gamma were used to calculate the fraction of aflatoxin contaminated kernels in contaminated lots. At 20 ppb, it was estimated that about 6 in 10,000 kernels are contaminated. PMID- 11048873 TI - Testing shelled corn for aflatoxin, Part III: evaluating the performance of aflatoxin sampling plans. AB - The effects of changes in sample size and/or sample acceptance level on the performance of aflatoxin sampling plans for shelled corn were investigated. Six sampling plans were evaluated for a range of sample sizes and sample acceptance levels. For a given sample size, decreasing the sample acceptance level decreases the percentage of lots accepted while increasing the percentage of lots rejected at all aflatoxin concentrations, and decreases the average aflatoxin concentration in lots accepted and lots rejected. For a given sample size where the sample acceptance level decreases relative to a fixed regulatory guideline, the number of false positives increases and the number of false negatives decreases. For a given sample size where the sample acceptance level increases relative to a fixed regulatory guideline, the number of false positives decreases and the number of false negatives increases. For a given sample acceptance level, increasing the sample size increases the percentage of lots accepted at concentrations below the regulatory guideline while increasing the percentage of lots rejected at concentrations above the regulatory guideline, and decreases the average aflatoxin concentration in the lots accepted while increasing the average aflatoxin concentration in the rejected lots. For a given sample acceptance level that equals the regulatory guideline, increasing the sample size decreases misclassification of lots, both false positives and false negatives. PMID- 11048874 TI - Sampling, sample preparation, and analytical variability associated with testing wheat for deoxynivalenol. AB - The variability associated with testing wheat for deoxynivalenol (DON) was measured using a 0.454 kg sample, Romer mill, 25 g comminuted subsample, and the Romer Fluoroquant analytical method. The total variability was partitioned into sampling, sample preparation, and analytical variability components. Each variance component was a function of the DON concentration and equations were developed to predict each variance component using regression techniques. The effect of sample size, subsample size, and number of aliquots on reducing the variability of the DON test procedure was also determined. For the test procedure, the coefficient of variation (CV) associated with testing wheat at 5 ppm was 13.4%. The CVs associated with sampling, sample preparation, and analysis were 6.3, 10.0, and 6.3%, respectively. For the sample variation, a 0.454 kg sample was used; for the sample preparation variation, a Romer mill and a 25 g subsample were used; for the analytical variation, the Romer Fluoroquant method was used. The CVs associated with testing wheat are relatively small compared to the CV associated with testing other commodities for other mycotoxins, such as aflatoxin in peanuts. Even when the small sample size of 0.454 kg was used, the sampling variation was not the largest source of error as found in other mycotoxin test procedures. PMID- 11048875 TI - Angiogenesis imaging. PMID- 11048876 TI - Accuracy of CT angiography versus pulmonary angiography in the diagnosis of acute pulmonary embolism: evaluation of the literature with summary ROC curve analysis. AB - RATIONALE AND OBJECTIVES: The authors performed this study to estimate, by using published data, the sensitivity and specificity of computed tomographic (CT) angiography in the evaluation of suspected acute pulmonary embolism (PE). MATERIALS AND METHODS: Summary receiver operating characteristic (ROC) curve analysis was used to determine the sensitivity and specificity of CT angiography in the diagnosis of acute PE. Pulmonary angiography was used as the diagnostic standard of reference. The authors reviewed the results of 11 independent studies published in the English-language literature between January 1992 and June 1999. RESULTS: The sensitivity of CT angiography in the diagnosis or exclusion of PE in the central pulmonary arteries (to the level of the segmental pulmonary arteries) ranged from 0.74 to 0.81 on the basis of specificities of 0.89-0.91. The sensitivity of CT angiography in the diagnosis or exclusion of PE in all pulmonary arteries (to the level of the subsegmental pulmonary arteries) was 0.68 on the basis of a specificity of 0.91. CONCLUSION: On the basis of the studies in the current literature, most of which used 5.0-mm collimation and single-detector CT, CT angiography may be less accurate in the diagnosis of PE than previously reported. With improvements in data acquisition, particularly the use of thinner section collimation and multidetector CT, and in the increased use of workstations for data analysis, the accuracy and utility of CT angiography will require continued investigation. PMID- 11048877 TI - Role of imaging in clinical trials of antiangiogenesis therapy in oncology. PMID- 11048878 TI - Tumor angiogenesis: molecular pathology, therapeutic targeting, and imaging. PMID- 11048879 TI - In vivo monitoring of tumor angiogenesis with MR imaging. PMID- 11048880 TI - Evaluation of tumor angiogenesis with US: imaging, Doppler, and contrast agents. PMID- 11048881 TI - Application of CT in the investigation of angiogenesis in oncology. PMID- 11048882 TI - Role of radionuclide imaging in trials of antiangiogenic therapy. PMID- 11048883 TI - Profile of medical student teaching in radiology: teaching methods, staff participation, and rewards. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to collect demographic information about radiology departments and rewards for teaching activities, as well as the impact of new digital imaging methods on teaching. MATERIALS AND METHODS: Two surveys were conducted of directors of medical school clerkships in radiology. The initial survey focused on numbers of staff and students, courses taught, and perception of rewards for teaching. The follow-up survey more specifically addressed teaching methods. RESULTS: Sixty-nine (50%) of the initial surveys sent to 139 departments and 46 (39% of a total of 119) of the follow-up surveys were returned. Clerkship directors spent an average of 9 hours per week teaching and performing administrative tasks, with most given no additional time off. Eighty-four percent of departments provide either no or insignificant rewards for teaching. Many departments have integrated the use of computers in teaching, and most have computers that students use during the radiology course. At the same time, digital imaging and picture archiving and communication systems (PACS) are used, or will be used within 1 year, in most departments. CONCLUSION: Clerkship directors receive little compensation in terms of time and rewards for medical student teaching. Teaching methods are evolving in response to the increasing use of computers, digital imaging, and PACS for at least part of the workload in most radiology departments. PMID- 11048884 TI - Readings in clinical imaging research: a structured bibliography. PMID- 11048885 TI - Molecular imaging in cancer: future directions and goals of the National Cancer Institute. PMID- 11048886 TI - In search of an endpoint. PMID- 11048887 TI - Role of extracellular retention of low density lipoproteins in atherosclerosis. AB - The pathogenesis for atherosclerosis is still unclear, and several hypotheses have been articulated to explain the initiating events in atherogenesis. Although these hypotheses are by no means mutually exclusive, there is a growing body of recent evidence that has led to the concept that subendothelial retention of apolipoprotein B100-containing lipoproteins is the initiating event in atherogenesis. Subsequently, a series of biological responses to this retained material leads to specific molecular and cellular processes that promote lesion formation. The present review assesses some of the studies that support this concept. PMID- 11048888 TI - Interaction of native and modified low-density lipoproteins with extracellular matrix. AB - Lipoprotein-matrix interactions play an important role in arterial disease. Extracellular matrix proteoglycans bind and retain specific positively charged domains on apolipoproteins B- and E-containing lipoproteins during atherogenesis. Retained lipoproteins can undergo several modifications, which may alter their interaction with extracellular matrix molecules. Growth factors, cytokines and oxidized low density lipoproteins influence proteoglycan structure, rendering them more likely to bind and retain lipoproteins during atherogenesis. Lipoproteins, native and modified, also can modulate the expression of several of the matrix degrading enzymes present in vascular tissue, thereby influencing plaque stability. Thus, the interaction of atherogenic lipoproteins with arterial wall matrix molecules can influence the genesis and progression of atherosclerosis and its complications. PMID- 11048889 TI - Phospholipase A2 and small, dense low-density lipoprotein. AB - High levels of small, dense LDL in plasma are associated with increased risk for cardiovascular disease. There are some biochemical characteristics that may render small, dense LDL particles more atherogenic than larger, buoyant LDL particles. First, small, dense LDL particles contain less phospholipids and unesterified cholesterol in their surface monolayer than do large, buoyant LDL particles. This difference in lipid content appears to induce changes in the conformation of apolipoprotein B-100, leading to more exposure of proteoglycan binding regions. This may be one reason for the high-affinity binding of small, dense LDL to arterial proteoglycans. Reduction of the phospholipid content in the surface monolayer LDL by treatment with secretory phospholipase A2 (sPLA2) forms small, dense LDL with an enhanced tendency to interact with proteoglycans. Circulating levels of sPLA2-IIA appears to be an independent risk factor for coronary artery disease and a predictor of cardiovascular events. In addition, in vivo studies support the hypothesis that sPLA2 proteins contribute to atherogenesis and its clinical consequences. These data suggest that modification of LDL by sPLA2 in the arterial tissue or in plasma may be a mechanism for the generation of atherogenic lipoprotein particles in vivo, with a high tendency to be entrapped in the arterial extracellular matrix. PMID- 11048890 TI - Metabolism of oxidized LDL by macrophages. AB - Oxidation products of lipids and proteins are found in atherosclerotic plaque and in macrophage foam cells. Macrophages avidly endocytose in-vitro oxidized LDL and accumulate sterols. What is the evidence that such a process is involved in in vivo foam cell formation? The present review surveys current knowledge on the metabolism of oxidized LDL by macrophages, and the types, amounts and location of oxidation products that accumulate in these cells. Comparable studies of lesion lipoproteins and foam cells indicate that limited extracellular lipoprotein oxidation, perhaps followed by more extensive intracellular oxidation subsequent to uptake by macrophages, is a more likely scenario in vivo. PMID- 11048891 TI - CD36 and atherosclerosis. AB - CD36 has been associated with diverse normal and pathologic processes. These include scavenger receptor functions (uptake of apoptotic cells and modified lipid), lipid metabolism and fatty acid transport, adhesion, angiogenesis, modulation of inflammation, transforming growth factor-beta activation, atherosclerosis, diabetes and cardiomyopathy. Although CD36 was identified more than 25 years ago, it is only with the advent of recent genetic technology that in-vivo evidence has emerged for its physiologic and pathologic relevance. As these in-vivo studies are expanded, we will gain further insight into the mechanism(s) by which CD36 transmits a cellular signal, and this will allow the design of specific therapeutics that impact on a particular function of CD36. PMID- 11048892 TI - ABC transporters in cellular lipid trafficking. AB - ATP-binding cassette (ABC) transporters constitute a group of evolutionary highly conserved cellular transmembrane transport proteins. Recent work has implicated ABC transporters in cellular transmembrane lipid transport and hereditary diseases have been causatively linked to defective ABC transporters translocating lipid compounds. The emerging concept that a defined subset of ABC transporters is intimately involved in cellular lipid trafficking has recently been substantiated convincingly by the finding that ABCA1 plays a central role in the regulation of HDL metabolism and macrophage targeting to the RES or the vascular wall. Differentiation dependent expression of a large number of ABC transporters in monocytes/macrophages and their regulation by sterol flux render these transporter molecules potentially critical players in atherogenesis and other chronic inflammatory diseases. PMID- 11048893 TI - Macrophage proliferation in atherosclerosis. AB - Oxidized LDL can induce an increase in intracellular calcium concentration and the activation of protein kinase C in mouse peritoneal macrophages. The activation of protein kinase C leads to the release into the culture medium of granulocyte-macrophage colony-stimulating factor, which plays a priming role in oxidized LDL-induced macrophage proliferation. The expression of granulocyte macrophage colony-stimulating factor in macrophages by oxidized LDL is positively regulated in the 5'-flanking region of granulocyte-macrophage colony-stimulating factor gene from sequence -169 to -160, but negatively regulated from -91 to -82. Granulocyte-macrophage colony-stimulating factor released by oxidized LDL from macrophages induces proliferation in autocrine or paracrine fashion via the activation of phosphatidylinositol 3-kinase. The capacity of oxidized LDL to induce macrophage proliferation in vitro may be involved in the enhanced progression of atherosclerosis in vivo. PMID- 11048894 TI - Dendritic cells and their involvement in atherosclerosis. AB - Dendritic cells constitute a unique family of cells able to induce primary immune responses. Over the past decade, immunologists have been increasingly preoccupied with dendritic cells and dendritic cells are now seen as a panacea for vaccine development, tumour immunotherapy and a host of other immunological applications. The recent finding of dendritic cells accumulating in atherosclerotic lesions should stimulate investigation of their contributions to atherogenesis and their potential use in anti-atherosclerosis therapies. PMID- 11048896 TI - Bibliography. Current world literature. Atherosclerosis: cell biology and lipoproteins. PMID- 11048895 TI - Cardiovascular disease in diabetes mellitus type 2: a potential role for novel cardiovascular risk factors. AB - A major consequence of diabetes mellitus type 2 is the accelerated development of atherosclerosis. Assessment of conventional risk factors such as plasma lipids, lipoproteins and hypertension only partly account for the excessive risk of developing cardiovascular disease in this population. Increasing evidence has emerged suggesting that conditions associated with diabetes mellitus type 2, such as insulin resistance, hyperinsulinemia and hyperglycemia, may also play a significant role in regulating 'novel' cardiovascular risk factors. These factors and their potential roles in the development of atherosclerosis and cardiovascular events are discussed in this review. PMID- 11048897 TI - Bimonthly update: lipidology. Nutrition and metabolism. PMID- 11048898 TI - Bimonthly update: lipidology. Genetics and molecular biology. PMID- 11048899 TI - Bimonthly update: lipidology. Hyperlipidaemia and cardiovascular disease. PMID- 11048900 TI - Bimonthly update: lipidology. Atherosclerosis: cell biology and lipoproteins. PMID- 11048901 TI - Consensus statement: anemia in HIV infection--current trends, treatment options, and practice strategies. Anemia in HIV Working Group. AB - BACKGROUND: Despite important advances in antiretroviral therapy, anemia remains a problem in many HIV-infected patients. Although the incidence of anemia in these patients has decreased, its prevalence appears to have stabilized or decreased only slightly. Anemia has a deleterious effect on both functional capacity and quality of life, and has been associated with shortened survival. OBJECTIVE: The Anemia in HIV Working Group, an expert panel of physicians and researchers involved in the care of HIV-infected patients, met to determine the impact of anemia in this patient population; to develop practice strategies for the clinician treating HIV-infected patients with anemia; and to identify future research directions. METHODS: The proposed practice strategies are based on results of the available clinical trials (as identified through a MEDLINE search), a review of the literature, and the clinical experience and expert opinion of the panel. The present report is based on meetings held in February and June of 1998; as further experience with various treatment options accumulates and the impact of highly active antiretroviral therapy becomes clearer, the panel will reconvene to develop evidence-based guidelines. RESULTS: The working group considers HIV-associated anemia to be an important contributor to the morbidity and mortality of this infection. Recent reports indicate that recovery from anemia is associated with improved quality of life and survival. CONCLUSIONS: As HIV-infected persons live longer, maintaining quality of life becomes an increasingly important goal of treatment. When planning treatment strategies, clinicians should consider the quality-of-life decrement caused by anemia. Transfusions should be used when rapid recovery is required, and underlying conditions causing anemia should be treated, if possible. Recombinant human erythropoietin (rHuEPO) therapy is appropriate in certain HIV-infected persons and should be considered to maintain hemoglobin concentrations. The target hemoglobin level is 12 g/dL for men and 11 g/dL for women. Weekly rHuEPO dosing is suggested, initiated at 40,000 U, as has been established in patients with cancer. PMID- 11048902 TI - The effectiveness of olanzapine in treatment-refractory schizophrenia when patients are nonresponsive to or unable to tolerate clozapine. AB - OBJECTIVE: This multicenter, open-label study was designed to assess the efficacy and tolerability of olanzapine in patients with chronic schizophrenia who are resistant to therapy with classic neuroleptic agents and are either not responsive to or unable to tolerate clozapine. METHODS: Patients received olanzapine orally once daily for 18 weeks at doses ranging from 5 to 25 mg. The primary efficacy measure was change in the total score on the Positive and Negative Syndrome Scale (PANSS) from baseline to end point. Secondary efficacy measures were the total score on the Brief Psychiatric Rating Scale (BPRS); the PANSS positive, negative, general psychopathology, and mood subscores; and the Clinical Global Impression improvement score. Also recorded were spontaneously reported adverse events; extrapyramidal symptoms (assessed by the Abnormal Involuntary Movement Scale, Simpson-Angus Scale, and Barnes Akathisia Scale); vital signs; and clinical laboratory test results. RESULTS: Forty-eight patients were treated with olanzapine; of these, 45 were assessable over the full 18-week study period. Total scores on the PANSS and BPRS were reduced from baseline by an average of 17.7 (14.2%) and 9.8 points (20.2%), respectively. Eighteen patients (40.0%) experienced a treatment response, defined as a reduction in PANSS total score of > or = 20%. A total of 25 patients (55.6%) achieved a similar reduction in BPRS total score. Significant reductions were seen in both the positive and negative symptom scores on the PANSS (P < 0.001). Olanzapine was well tolerated, with minimal treatment-emergent adverse events or clinically relevant changes in vital signs or clinical laboratory test results. No clinically significant blood dyscrasias were observed in olanzapine-treated patients, including those who had discontinued clozapine because of treatment-associated leukopenia or neutropenia. CONCLUSION: The results of this study suggest that olanzapine may be of benefit in patients who are refractory to or unable to tolerate clozapine. PMID- 11048903 TI - Effect of early intervention with sumatriptan on migraine pain: retrospective analyses of data from three clinical trials. AB - OBJECTIVE: This study assessed the efficacy of sumatriptan 50- and 100-mg tablets in the treatment of migraine attacks while the pain is mild rather than moderate/severe. BACKGROUND: Results from The Spectrum Study suggested that early treatment of migraine attacks with sumatriptan 50-mg tablets while the pain is mild might enhance pain-free response and reduce headache recurrence. METHODS: Retrospective analyses of headaches treated during mild pain were performed using data from 3 studies of sumatriptan tablets (protocols S2CM09, S2BT25, and S2BT26). Our primary interest was pain-free response 2 and 4 hours after dosing; secondary interests were use of a second dose of medication, clinical disability (as measured on a 4-point disability scale), migraine-associated symptoms, meaningful pain relief (patient defined), time to meaningful relief, sustained pain-free response, and proportion of attacks in which pain had worsened 2 and 4 hours after dosing, all of which were compared in headaches treated during mild versus moderate/severe pain. RESULTS: In S2CM09, 92 patients treated 118 headaches during mild pain. Rates of pain-free response were higher 2 hours after dosing with sumatriptan 50 mg (51%) or 100 mg (67%; P < 0.05) compared with placebo (28%), and were higher with early treatment of mild pain compared with treatment of moderate/severe pain at 2 hours (sumatriptan 50 mg: mild pain, 51%; moderate/severe pain, 31%; P < 0.05; sumatriptan 100 mg: mild pain, 67%; moderate/severe pain, 36%) and 4 hours (50 mg: 75% vs 56%; 100 mg: 90% vs 61%; P < 0.05). Early intervention also resulted in less redosing than when moderate/severe pain was treated (50 mg: 21% vs 32%; 100 mg: 20% vs 29%). More attacks treated early with sumatriptan 50 or 100 mg were associated with normal function 4 hours after dosing compared with placebo (70% and 93% vs 46%, respectively). Sustained pain-free response rates 2 to 24 hours after early dosing with sumatriptan 50 or 100 mg were also higher (34% and 53%, respectively) compared with treatment of moderate/severe pain (19% and 24%, respectively). Early treatment with sumatriptan 100 mg produced significantly higher pain-free rates at 2 hours after dosing (P < 0.001) than did ergotamine plus caffeine (S2BT25: 69% vs 34%, respectively) or aspirin plus metoclopramide (S2BT26: 73% vs 25%, respectively). CONCLUSIONS: Sumatriptan 50- and 100-mg tablets are effective whether pain is mild or moderate/severe. However, treatment with sumatriptan while pain is mild provides high pain-free response rates while reducing the need for redosing, benefits not seen with ergotamine plus caffeine or aspirin plus metoclopramide. PMID- 11048904 TI - Patient and physician satisfaction with the Humulin/Humalog Pen, a new 3.0-mL prefilled pen device for insulin delivery. AB - OBJECTIVE: This study assessed acceptability of a new 3.0-mL prefilled insulin pen device, the Humulin/Humalog Pen, as a method of delivering human insulin. Secondary objectives were to determine whether the pen device might facilitate initiation of insulin therapy in patients currently receiving oral antihyperglycemic agents and to monitor the safety of this pen device in clinical practice. BACKGROUND: For both the patient and health care provider, significant negative perceptions of the use of insulin therapy persist, including patient inconvenience, social stigma from insulin injections, and insufficient time for the provider to train the patient. METHODS: This 6-week, open-label, noncomparative study was conducted at 33 centers in the United States. Patients with type 1 or type 2 diabetes treated with insulin therapy or oral antihyperglycemic agents were enrolled in the study. Before the study, 62% (194 patients) had used a syringe and vial for insulin injection, 28% (87 patients) had used an insulin pen device, and 10% (30 patients) were insulin-naive. Prior therapy was unknown in 1% (4 patients). Patients used the Humulin/Humalog Pen for > or = 1 injection of insulin daily for 6 weeks. At the beginning and end of the study, patients completed a questionnaire designed to elicit their perceptions of the Humulin/Humalog Pen; physicians completed a questionnaire at the end of the study. Frequencies and percentages of all categoric responses were calculated and summarized. RESULTS: A total of 315 patients (136 type 1, 179 type 2 diabetes) were enrolled. Of the 299 patients who completed questionnaires at the end of the study, 76% (226 patients) were somewhat or extremely satisfied with the pen, 78% (234 patients) probably or definitely would continue to use the pen, and 80% (239 patients) probably or definitely would recommend the pen to others. Of the 33 physicians who completed questionnaires at the end of the study, 97% (32) thought that the pen was better overall compared with a vial and syringe, 88% (29) thought that it took less time to teach patients to use the pen, and 73% (24) thought that it took less time to initiate insulin therapy with the pen. CONCLUSIONS: The Humulin/Humalog Pen had an acceptable safety profile and was well accepted by patients and physicians. PMID- 11048905 TI - Long-Term use of quetiapine in elderly patients with psychotic disorders. AB - BACKGROUND: Quetiapine is an atypical antipsychotic agent that does not appear to increase patient risk for treatment-emergent extrapyramidal symptoms (EPS) or anticholinergic symptoms. Previous studies of quetiapine use in elderly patients with schizophrenia and other psychoses examined short-term administration (< or = 12 weeks). Given the growing elderly population, the commensurate increase in elderly patients with psychoses, and the expected increase in disease treatment years, the effect of long-term quetiapine administration in older patients is of considerable interest. OBJECTIVE: This study assesses the long-term tolerability, safety, and clinical benefit of quetiapine in elderly patients with psychosis. METHODS: Elderly patients (> or = 65 years of age) with psychotic disorders, as defined by the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, participated in this 52-week, open-label, multicenter trial. Investigators increased (and later adjusted) daily doses of quetiapine on the basis of clinical response and tolerability, and assessed safety and efficacy. Efficacy assessments were made using the 18-item Brief Psychiatric Rating Scale (BPRS), Clinical Global Impressions (CGI), Simpson-Angus Scale, and the Abnormal Involuntary Movement Scale (AIMS). For patients who withdrew before week 52, analyses were performed using observed data and the last observation carried forward. RESULTS: One hundred eighty-four patients with psychotic disorders (98 women and 86 men) with a mean age of 76.1 years entered the trial. Seventy-two percent had psychotic disorders due to general medical conditions such as Alzheimer's disease, and 28% had other psychotic disorders, most commonly schizophrenia. Overall, 89 (48%) patients completed treatment through 52 weeks. Median total daily dose was 137.5 mg. Reasons, for withdrawal included lack of efficacy (19%), adverse events or intercurrent illness (15%), failure to return for follow-up (13%), protocol noncompliance (3%), and diminished need for treatment (2%). Somnolence (31%), dizziness (17%), and postural hypotension (15%) were common adverse events, but they rarely resulted in withdrawal from therapy. EPS-related adverse events occurred in 13% of patients. At end point (week 52), mean total score on the Simpson-Angus Scale had decreased from baseline by 1.8 points, whereas changes in AIMS scores were negligible. No clinically important effects were reported relative to mean changes in hematologic, thyroid function, or hepatic function variables. Quetiapine treatment appeared to have no associated cardiovascular adverse outcomes despite cardiovascular comorbidities and unrestricted use of concomitant cardiovascular medications. Significant decreases in BPRS total score (n = 170, P < 0.001) and CGI Severity of Illness item score (n = 177, P < 0.002) were seen at end point (observed data and last observation carried forward). Decreases of > or = 20% in mean BPRS total score were observed in 83 (49%) patients. CONCLUSIONS: These results provide preliminary information to clinicians regarding tolerability, safety, and clinical improvement with quetiapine in elderly patients with psychotic symptoms, and support controlled studies of quetiapine in this patient population. PMID- 11048906 TI - The effects of olanzapine, risperidone, and haloperidol on plasma prolactin levels in patients with schizophrenia. AB - BACKGROUND: There is relatively little comparative information on elevations in plasma prolactin level (PRL) with conventional versus novel antipsychotic agents. OBJECTIVE: This paper examines the comparative effects on PRL of olanzapine, risperidone, and haloperidol based on data from 3 multicenter, double-blind, randomized clinical trials. Magnitude of response, dose dependency, time course, effects of sex and age, and response to switching from haloperidol to olanzapine are assessed. METHODS: The effects of olanzapine, risperidone, and haloperidol on PRL were assessed in patients with schizophrenia or related psychoses participating in 3 double-blind clinical trials: (1) a 6-week acute trial comparing olanzapine 5 to 20 mg/d (n = 1,336) and haloperidol 5 to 20 mg/d (n = 660), with a 1-year, open-label olanzapine extension for responders; (2) a 54 week study comparing olanzapine 5 to 20 mg/d (n = 21), risperidone 4 to 10 mg/d (n = 21), and haloperidol 5 to 20 mg/d (n = 23) in early illness; and (3) a 28 week study comparing olanzapine 10 to 20 mg/d (n = 172) and risperidone 4 to 12 mg/d (n = 167). RESULTS: PRL elevations were significantly greater with risperidone than with either olanzapine or haloperidol in study 2. and significantly greater than with olanzapine in study 3 (all, P < 0.001). PRL elevations were significantly greater with haloperidol than with olanzapine in study 1 (P < 0.001 ). A dose-response relationship was not consistently confirmed with any of the drug treatments. Risperidone-associated PRL elevations peaked relatively early in treatment. In haloperidol- and risperidone-treated patients, the mean change in PRL was greater in women than in men. PRL decreased significantly when treatment was switched from haloperidol to olanzapine. CONCLUSIONS: This side-by-side analysis of 3 independent studies suggests that with the 3 antipsychotic drugs studied, PRL is elevated moderately by olanzapine (mean change, 1-4 ng/mL), intermediately by haloperidol (mean change, approximately 17 ng/mL), and strongly by risperidone (mean change, 45-80 ng/mL). No consistent dose-response relationship was observed, and the time course and sex-dependency of the response differed between the 3 agents. Patients with haloperidol-induced hyperprolactinemia may benefit from a switch to olanzapine. Long-term studies examining the health consequences of chronic hyperprolactinemia during antipsychotic treatment are needed. PMID- 11048907 TI - Development of subscales from the symptoms/problems and effects of kidney disease scales of the kidney disease quality of life instrument. AB - BACKGROUND: The Kidney Disease Quality of Life Instrument (KDQOL) was developed to provide clinicians with a comprehensive assessment of the important domains of health-related quality of life (HRQOL) for patients with end-stage renal disease who are undergoing hemodialysis. OBJECTIVE: The purpose of this study was to develop subscales from the 55 items comprising the Symptoms/Problems and Effects of Kidney Disease scales of the KDQOL and to measure the internal consistency reliability of these subscales. METHODS: The 55 items from the Symptoms/Problems and Effects of Kidney Disease scales were arranged into substantively meaningful clusters using an affinity mapping procedure. The resulting subscales were assessed for internal consistency reliability using data from a sample of 165 individuals with kidney disease who had completed the KDQOL. RESULTS: Eleven multi-item subscales were identified: pain, psychological dependency, cognitive functioning, social functioning, dialysis-related symptoms, cardiopulmonary symptoms, sleep, energy, cramps, diet, and appetite. Four items (clotting or other problems with access site, high blood pressure, numbness in hands or feet, and blurred vision) were not included in any of these subscales. Internal consistency reliability estimates for the 11 subscales ranged from 0.66 to 0.92. These subscales correlated with the scales from the 36-Item Short-Form Health Survey as hypothesized (ie, corresponding pain, energy, and social functioning scales had the highest correlations). In addition, several subscales were significantly associated, as hypothesized, with other variables such as the number of disability days. CONCLUSIONS: The results of this study further support the reliability and validity of the KDQOL. The 11 subscales identified yield more detailed information on the HRQOL of patients with kidney disease and provide a basis for specific improvements in the quality of care delivered to these patients. PMID- 11048908 TI - The discriminative ability of the 12-item short form health survey (SF-12) in a sample of persons infected with HIV. AB - BACKGROUND: Self-reported health-related quality of life (HRQOL) assesses constructs that transcend laboratory-based clinical parameters. Corroboration of the hypothesized relationships between the 2 types of health indicators (ie, clinical and HRQOL) could provide evidence of the validity of an HRQOL measurement tool. OBJECTIVE: The purpose of this study was to evaluate the ability of scores on the mental component summary (MCS-12) and physical component summary (PCS-12) of the 12-Item Short Form Health Survey (SF-12) to discriminate between HIV-infected persons in predefined disease-severity groups based on surrogate markers. METHODS: This cross-sectional study involved the collection of clinical data (ie, CD4 cell count, viral load [HIV-1 RNA copies/mL]) from patients' medical records and HRQOL data from the SF-12 at 2 HIV specialty clinics. The ability of SF-12 summary scores to discriminate between patients stratified by disease severity (ie, CD4 cell count <200 vs > or = 200/mm3; HIV-1 RNA >55,000 vs < or = 55,000 copies/mL) was assessed by receiver operating characteristic curve analysis. RESULTS: Data were collected from 478 patients. The scores from the PCS-12 were able to discriminate between groups of patients stratified by disease severity based on CD4 cell count (P < 0.001) and HIV-1 RNA copies/mL (P < 0.01). MCS-12 scores did not discriminate between disease-severity groups. CONCLUSIONS: Although the SF-12 is a brief generic measure of HRQOL, these findings provide further evidence of the validity of the SF-12 and suggest that it may be a practical way to monitor health status from the perspective of the HIV-infected patient. PMID- 11048909 TI - American translation, modification, and validation of the St. George's Respiratory Questionnaire. AB - BACKGROUND: The St. George's Respiratory Questionnaire (SGRQ) is a 50-item health status survey specific for chronic obstructive pulmonary disease (COPD) and other respiratory diseases that is available in British English but not American English. The SGRQ's symptom-reporting component requires a 1-year reporting period, which may be too long for reliable and accurate patient recall. OBJECTIVES: The objectives of the present study were to translate the SGRQ from British to American English, modify the reporting period of the symptom-reporting component from 1 year to 1 month, and assess the reliability, validity, and sensitivity to change of this translated modified version in a sample of patients with COPD. METHODS: Based on input from American patients with COPD and health professionals, the SGRQ was translated into American English (SGRQ-A) and then translated back to British English. For SGRQ-A reliability and validity studies, patients were asked to report symptoms experienced over 1 year (reporting period in the original SGRQ) and 1 month (modification made to SGRQ-A). We evaluated 102 patients with COPD (50% female; mean age, 68 years; mean forced expiratory volume in 1 second [FEV1], 1.01 L) at an administrative session before and after completion of a pulmonary rehabilitation program. The SGRQ-A, Chronic Respiratory Disease Questionnaire (CRQ), 36-Item Short Form Health Survey (SF-36), 6-minute walk (6MW), Medical Research Council (MRC) Dyspnea scale, and pulmonary function tests (FEV1 and % predicted FEV1) were used in the assessment battery. RESULTS: The SGRQ-A showed good agreement with the original SGRQ when translated back to British English. Internal reliability (Cronbach alpha) was > 0.70 for all SGRQ-A components except the 1-year symptom-reporting component. Test-retest intraclass correlations were 0.795 to 0.900. Construct validity was strengthened when all SGRQ-A components (except 1-year symptoms and most 1-month symptoms) correlated (P < or = 0.01) with the MRC Dyspnea scale, 6MW, all SF-36 concept scores, and 80% of CRQ domains (r = 0.30-0.72). Discriminate validity was demonstrated when all components of the SGRQ-A with the modified 1-month symptom-reporting period were shown to discriminate better between disease-severity groups (based on patient self-reports of disease severity) than did pulmonary function tests and the 6MW. Responsiveness of the SGRQ-A to change in health status was demonstrated when scores on the Symptoms-1 month and Total-1 month components detected significant improvements in patients' health status (P = 0.02 and P = 0.04, respectively). CONCLUSION: The SGRQ-A with a modified 1-month symptom-reporting period demonstrated reliability and validity in this sample of patients with COPD. Key words: chronic obstructive pulmonary disease, St. George's Respiratory Questionnaire, American translation, reliability, validity, symptom recall. PMID- 11048910 TI - Use of lamotrigine in a patient with bipolar disorder and psychiatric comorbidity. PMID- 11048911 TI - Growth-promoting systems for heifer calves and yearlings finished in the feedlot. AB - In a 172-d finishing trial (Exp. 1), 210 recently weaned crossbred heifers were allotted to six growth promotant treatment groups, involving implanting initially with Synovex-C (C) or H (H) followed by reimplanting with Finaplix-H (F) or H and F. Melengestrol acetate (MGA) was provided in the diet to four of the treatment groups. Heifers fed MGA and administered only F as the terminal implant had the greatest (P = .01) number of mature ovaries with follicles but also had lower (P = .01) gain/DMI. In a 182-d finishing study (Exp. 2), 270 recently weaned crossbred heifers were allotted to the following six implant (d 0)/ reimplant (d 70) groups using no implant (N), Ralgro (R) or H: N/R, R/H, R/R, N/R, H/H and R/R for Treatments 1 through 6, respectively. On d 70, all heifers were implanted with F. Heifers were fed MGA from d 70 to 182 (Treatments 1, 2, and 3) or for the entire trial (Treatments 4, 5, and 6). Implanting on d 0 increased (P < .05) overall ADG. Differences (P > .05) in performance were not found between MGA treatment groups. Using an H implant/reimplant regimen decreased (P = .01) ovarian and(or) follicular development when compared with an R implant/reimplant regimen. In a 126-d finishing trial (Exp. 3), 360 crossbred yearling heifers were used to evaluate F and estrogen (Implus-H) implants when used in combination with an MGA feeding program. Heifers receiving only F in combination with MGA had greater (P < .05) ADG, whereas all heifers fed MGA had greater (P < .05) gain/DMI than heifers not fed MGA. These data suggest that feeding MGA was not beneficial for young heifers, particularly if they are provided an initial estrogenic implant followed by a second implant. In older (yearling) heifers, increased gains and gain/DMI were obtained by feeding MGA and implanting initially or 56 d later with F. PMID- 11048912 TI - Effects of ruminally protected choline and dietary fat on performance and blood metabolites of finishing heifers. AB - A 120-d finishing study utilizing 318 heifers (342 kg initial BW) was conducted to examine effects of ruminally protected choline (RPC) in diets containing graded concentrations of tallow. Heifers were blocked according to previous nutrition (full-fed or limit-fed) and allotted to 24 pens containing 11 to 15 heifers. Two pens, one within each block, were assigned to each of 12 factorially arranged treatments including dietary tallow (0, 2, or 4%) and supplemental RPC (0, 20, 40, or 60 g of product daily, estimated to supply 0, 5, 10, or 15 g/d choline postruminally). Heifers were implanted with Revalor-H and fed a finishing diet based on steam-flaked and dry-rolled corn (12.5% CP, 8% alfalfa on DM basis). Dry matter intake decreased (P < 0.10) by 5.4% when tallow was increased from 0 to 4% but was not affected by RPC. Heifers receiving 4% tallow had 7.3% lower gains than those receiving none (P < 0.10). Supplementation of RPC increased (P < 0.10) ADG, with 20 g/d resulting in an 8.6% increase. Similarly, gain efficiency improved (P < 0.10) by 7.6% with addition of 20 g/d RPC. Yield grade and kidney, pelvic, and heart fat both increased linearly (P < 0.10) with fat supplementation. The percentage of carcasses grading USDA Choice was not affected by intermediate levels of RPC but decreased with the highest level (60 g/d). Dressing percentage, hot carcass weight, marbling, and 12th-rib fat thickness were not affected significantly by either tallow or RPC. On d 90, jugular blood was collected from all heifers at 2 h postfeeding. Plasma urea and serum insulin concentrations were not affected by either tallow or RPC. Dietary tallow linearly increased (P < 0.10) NEFA, cholesterol, triglyceride, and total amino acid concentrations. Choline supplementation led to quadratic responses for total amino acids (P < 0.10), with concentrations being greatest for intermediate levels of RPC. Moderate levels of supplemental RPC improved growth performance of finishing cattle without negatively affecting carcass characteristics. Optimum performance was achieved with 20 g of product daily. PMID- 11048913 TI - Growth performance and adipose tissue deposition in barrows fed n-methyl-D,L aspartate. AB - We previously reported that broilers fed n-methyl-D,L-aspartate (NMA) exhibited enhanced feed conversion efficiency and decreased percentage of fat in carcasses. In this experiment, growth performance and backfat thickness were evaluated in barrows fed NMA. Poland China x Yorkshire barrows weighing 68.8 +/- 1.7 kg (mean +/- SE) were allowed ad libitum access to feed containing NMA at levels of either 0 (n = 7), 100 (n = 6), 200 (n = 8), or 300 (n = 8) mg/kg for 36 d. Barrows were slaughtered at 99.5 +/- 2.3 kg BW. There was no effect (P > 0.1) of NMA on ADG or feed consumption. Gain:feed ratio decreased (P < 0.03) in a linear fashion with increasing level of NMA. There was a cubic effect (P < 0.05) of NMA treatment on first-rib backfat thickness. In response to graded levels of NMA, backfat thickness at the 10th rib (P < 0.08) and last rib (P < 0.03) increased in a linear fashion. The NMA had no effect (P > 0.1) on backfat thickness measured at the lumbar vertebra or longissimus muscle area measured at the 10th rib interface. The percentage of lean in the carcass decreased in a linear fashion (P < 0.05) in response to increasing levels of NMA in the diet. In summary, NMA had an overall negative effect on growth performance and carcass yield characteristics in barrows. The dichotomous effects of NMA on feed efficiency and body composition in poultry and swine warrants further scrutiny. PMID- 11048914 TI - Effects of intramammary infection and parity on calf weaning weight and milk quality in beef cows. AB - The objectives of this study were to determine 1) the effect of intramammary infection on calf weaning weight, milk somatic cell count, and milk composition, and 2) the effect of parity on percentages of infected cows, infected quarters, and blind quarters. The number of infected quarters, milk somatic cell counts, milk components, and intramammary infection were studied at weaning in 164 beef cows. The percentage of infected cows ranged from 61.9% at first parity to 66.7% at fifth to ninth parities. Cows with three or four infected quarters had higher (P < .01) milk somatic cell counts than cows with zero, one, or two infected quarters. Among bacterial isolates, Staphylococcus aureus-infected quarters had the highest (P < .01) milk somatic cell count. Percentages of butterfat and lactose were lower (P < .01) in milk from infected quarters than from uninfected quarters. Infections by S. aureus and coagulase-negative staphylococci were the most common and accounted for 67 to 78% of the infections. Percentages of infected quarters and infections caused by S. aureus increased with parity (P < .01). Intramammary infections did not affect (P > .10) calf weaning weight. In conclusion, intramammary infection had no effect on calf weaning weight but increased milk somatic cell count and decreased the percentage of protein, lactose, solids-not-fat, and butterfat. The number of infected and blind mammary quarters increased with parity. PMID- 11048915 TI - Genetic analysis of growth curve parameters for male and female chickens resulting from selection on shape of growth curve. AB - The objectives of this research were to evaluate gender differences and selection on body weight as they affect growth curves of chickens. Marginal posterior densities of growth curve parameters were studied by Gibbs sampling on 10,671 male and female chickens originating from five lines. Line X-+ was selected on low body weight at 8 wk (BW8) and high body weight at 36 wk (BW36), line X+- on high BW8 and low BW36, X++ on high BW8 and BW36, X-- on low BW8, and BW36, and X00 was an unselected control line. Growth was modeled by a Gompertz function. Heritabilities and genetic correlations among parameters of the Gompertz curve were estimated. Marginal posterior densities were drawn for parameters of the growth curve and for sexual dimorphism at ages ranging from hatching to 1 yr. Lines selected for a higher BW8 had higher initial specific growth rates (L), higher maturation rates (K), and lower ages at inflection (T(I)). Lines selected for a higher BW36 had higher asymptotic body weights (A). Estimates of A, L, and T(I) were higher in males and K was higher in females. Difference between sexes for A was greater in the line selected for a lower BW8 and a higher BW36. Dimorphism for L and K was the lowest in lines X++ and X--, respectively. The greatest difference in T(I) was observed in the line selected for lower BW8 and BW36. Sexual dimorphism of body weight was lower at most ages in the lightest line. Before 15 wk, sexual dimorphism in X++ line was lower than in the line selected for higher BW8 and lower BW36. The increase in sexual dimorphism with body weight could be reduced by selecting animals on body weight at two ages instead of one, as is usually done in commercial lines. PMID- 11048917 TI - Genetic parameters for direct and maternal calving ability over parities in Piedmontese cattle. AB - Estimates of heritabilities and genetic correlations for calving ease over parities were obtained for the Italian Piedmontese population using animal models. Field data were calving records of 50,721 first- and 44,148 second-parity females and 142,869 records of 38,213 cows of second or later parity. Calving ability was scored in five categories and analyzed using either a univariate or a bivariate linear model, treating performance over parities as different traits. The bivariate model was used to investigate the genetic relationship between first- and second- or between first- and third-parity calving ability. All models included direct and maternal genetic effects, which were assumed to be mutually correlated. (Co)variance components were estimated using restricted maximum likelihood procedures. In the univariate analyses, the heritability for direct effects was .19 +/- .01, .10 +/- .01, and .08 +/- .004 for first, second, and second and later parities, respectively. The heritability for maternal effects was .09 +/- .01, .11 +/- .01, and .05 +/- .01, respectively. All genetic correlations between direct and maternal effects were negative, ranging from -.55 to -.43. Approximated standard errors of genetic correlations between direct and maternal effects ranged from .041 to .062. For multiparous cows, the fraction of total variance due to the permanent environment was greater than the maternal heritability. With bivariate models, direct heritability for first parity was smaller than the corresponding univariate estimate, ranging from .18 to .14. Maternal heritabilities were slightly higher than the corresponding univariate estimates. Genetic correlation between first and second parity was .998 +/- .00 for direct effects and .913 +/- .01 for maternal effects. When the bivariate model analyzed first- and third-parity calving ability, genetic correlation was .907 +/- .02 for direct effects and .979 +/- .01 for maternal effects. Residual correlations were low in all bivariate analyses, ranging from .13 for analysis of first and second parity to .07 for analysis of first and third parity. In conclusion, estimates of genetic correlations for calving ease in different parities obtained in this study were very high, but variance components and heritabilities were clearly heterogeneous over parities. PMID- 11048916 TI - A QTL on pig chromosome 4 affects fatty acid metabolism: evidence from an Iberian by Landrace intercross. AB - Three Iberian boars were bred to 31 Landrace sows to produce 79 F1 pigs. Six F1 boars were mated to 73 F1 sows. The F2 progeny from 33 full-sib families (250 individuals) were genotyped for seven microsatellites spanning the length of chromosome 4. Least squares procedures for interval mapping were used to detect quantitative trait loci (QTL). A permutation test was used to establish nominal significance levels associated with QTL effects, and resulting probability levels were corrected to a genomewide basis. Observed QTL effects were (genomewide significance, position of maximum significance in centimorgans): percentage of linoleic acid in subcutaneous adipose tissue (< 0.01, 81); backfat thickness (< 0.01, 83); backfat weight (< 0.01, 80); longissimus muscle area (0.02, 83); live weight (0.19, 88); and percentage of oleic acid in subcutaneous adipose tissue (0.25, 81). Gene action was primarily additive. The Iberian genotypes were fatter, slower growing, and had lower linoleic and higher oleic acid contents than Landrace genotypes. The interval from 80 to 83 cM contains the FAT1 and A FABP loci that have been shown previously to affect fat deposition in pigs. This is the first report of a QTL affecting fatty acid composition of subcutaneous adipose tissue in pigs and provides a guide for the metabolic pathways affected by candidate genes described in this region of chromosome 4. PMID- 11048918 TI - Relationship between a bull's parental genetic merit difference and subsequent progeny trait variability in Angus, Gelbvieh, and Limousin cattle. AB - Data from the American Angus Association, American Gelbvieh Association, and the North American Limousin Foundation were analyzed to determine whether parental genetic differences are associated with Mendelian sampling of their bull progeny or with Mendelian sampling variances and weight variances of their bull progeny's offspring. Parental differences were measured as the difference between the parents' EPD for birth weight (DIF(BW)), weaning weight direct (DIF(WW)), and yearling weight (DIF(YW)). A bull's data were used if both parents had calculated EPD and the bull had at least 25 progeny with records for the specific trait. Traits calculated for each bull were his Mendelian sampling (MS(Bull)), progeny Mendelian sampling variance (MSsigma2progeny), progeny weight variance (WTsigma2), and progeny corrected weight variance (CWTsigma2 = adjusted weight minus appropriate dam EPD) for birth, weaning, and yearling weights. Pearson correlations were computed between DIF(BW), DIF(WW), and DIF(YW) and MS(Bull), MSsigma2progeny, WTsigma2, and CWTsigma2 for each trait, within each breed. Across breeds, the correlations ranged from -.07 to .11 for MS(Bull) .01 to .14 for MSsigma2progeny, -.06 to .09 for WTsigma2, and -.06 to .08 for CWTsigma2. Although some of the correlations were significantly different from zero their relatively small magnitude indicates little relationship between parental differences in genetic merit and subsequent offspring variability for each of the three breeds. PMID- 11048919 TI - Identification of quantitative trait loci affecting birth characters and accumulation of backfat and weight in a Meishan-White Composite resource population. AB - A search for genomic regions affecting birth characters and accretion of weight and backfat was conducted in a Meishan-White Composite reciprocal backcross resource population. Birth traits analyzed (n = 750) were vigor score, number of nipples, and birth weight. Subsequent measures on gilts and barrows (n = 706) analyzed were weaning weight, 8-wk weight, ADG from 8 to 18 wk of age, ADG from 18 to 26 wk of age, 26-wk weight, and backfat over the first rib, last rib, and last lumbar vertebrae at 14 and 26 (n = 599) wk of age. Feed intake and growth of 92 individually penned barrows were also analyzed. A genomic scan was conducted with microsatellite markers spaced at approximately 20-cM intervals, a least squares regression interval analysis was implemented, and significance values were converted to genomewide levels. No associations were detected for traits measured at birth except for number of nipples, where one significant and two suggestive regions were identified on chromosomes (SSC) 10, 1, and 3, respectively. Early growth was affected by a region on SSC 1 as evidenced by associations with weights collected at weaning and 8 wk of age and ADG from 8 to 18 wk of age. Other regions detected for early growth rate were on SSC 2, 12, and X. Chromosomal regions on SSC 6 and 7 affected ADG from 18 to 26 wk of age. All measures of backfat were affected by regions on SSC 1 and X, whereas SSC 7 consistently affected backfat measures recorded at 26 wk of age. Suggestive evidence for QTL affecting backfat at 14 wk of age was also detected on SSC 2, 6, 8, and 9. These results have improved our knowledge about the genetics of growth rate and fat accretion at the molecular level in swine. PMID- 11048920 TI - Comparison of restricted maximum likelihood and method R for estimating heritability and predicting breeding value under selection. AB - Method R and Restricted Maximum Likelihood (REML) were compared for estimating heritability (h2) and subsequent prediction of breeding values (a) with data subject to selection. A single-trait animal model was used to generate the data and to predict breeding values. The data originated from 10 sires and 100 dams and simulation progressed for 10 overlapping generations. In simulating the data, genetic evaluation used the underlying parameter values and sires and dams were chosen by truncation selection for greatest predicted breeding values. Four alternative pedigree structures were evaluated: complete pedigree information, 50% of phenotypes with sire identities missing, 50% of phenotypes with dam identities missing, and 50% of phenotypes with sire and dams identities missing. Under selection and with complete pedigree data, Method R was a slightly less consistent estimator of h2 than REML. Estimates of h2 by both methods were biased downward when there was selection and loss of pedigree information and were unbiased when no selection was practiced. The empirical mean square error (EMSE) of Method R was several times larger than the EMSE of REML. In a subsequent analysis, different combinations of generations selected and generations sampled were simulated in an effort to disentangle the effects of both factors on Method R estimates of h2. It was observed that Method R overestimated h2 when both the sampling that is intrinsic in the method and the selection occurred in generations 6 to 10. In a final experiment, BLUP(a) were predicted with h2 estimated by either Method R or REML. Subsequently, five more generations of selection were practiced, and the mean square error of prediction (MSEP) of BLUP(a) was calculated with estimated h2 by either method, or the true value of the parameter. The MSEP of empirical BLUP(a) using Method R was greater than the MSEP of empirical BLUP(a) using REML. The latter statistic was closer to prediction error variance of BLUP(a) than the MSEP of empirical BLUP(a) using Method R, indicating that empirical BLUP(a) calculated using REML produced accurate predictions of breeding values under selection. In conclusion, the variability of h2 estimates calculated with Method R was greater than the variability of h2 estimates calculated with REML, with or without selection. Also, the MSEP of EBLUP(a) calculated using estimates of h2 by Method R was larger than MSEP of EBLUP(a) calculated with REML estimates of h2. PMID- 11048921 TI - Immune status of PIC Camborough-15 sows, 25% Meishan sows, and their offspring kept indoors and outdoors. AB - Newer genetic lines of pigs are being used in indoor and outdoor production systems. The objectives of Exp. 1 were to describe the effects of the maternal sow line genotype, environment (indoor vs outdoor), and the genotype x environment interactions on blood hemoglobin (Hb), immunoglobulin G (IgG) concentrations, white blood cell (WBC) numbers, lymphocyte transformation/blastogenesis (LTA), natural killer (NK) cell activity, neutrophil chemotaxis, cortisol concentrations, and leukocyte differentials. Studies were performed using two genotypes: PIC Experimental-94 (Exp-94, an experimental line containing 25% Meishan) and PIC Camborough-15 (C-15). The Exp-94 sows had lower LTA at 0.2 microg/mL mitogen than the C-15 sows, whereas Exp-94 sows had higher NK cytotoxicity than the C-15 sows. When indoors, the two genotypes showed similar neutrophil chemotaxis. When outdoors, the C-15 genotype had higher (P < .01) neutrophil chemotaxis than the Exp-94 sows. The other immune measures were statistically similar for the two genotypes for each environment and for the genotype x environment interaction of sows. Experiment 2 sought to determine the effects of genotype on the immune system of nursery-age offspring of the experimental lines. Each sow line was bred to a common PIC 405 boar line. The Exp 94 x 405 pigs had elevated WBC numbers than C-15 x 405 pigs. The social status of the Exp-94 x 405 or the C-15 x 405 pigs showed no effect on any of the immune measures studied. The other immune measures were statistically similar for the two lines of pigs. The Exp-94 line had marginally increased NK activity but reduced lymphocyte blastogenesis and neutrophil chemotaxis compared with the C-15 line. PMID- 11048922 TI - Dehydration, stress, and water consumption of horses during long-distance commercial transport. AB - The aim of this study was to characterize progressive dehydration, stress responses, and water consumption patterns of horses transported long distances in hot weather and to estimate recovery time after 30 h of transport. Thirty adult mares and geldings were deprived of access to feed and water for 6 h, blocked by age, sex, breed, and body condition score, and assigned to one of the following treatments: penned, offered water (Penned/Watered, n = 5); penned, no water (Penned, n = 5); transported, offered water (Transported/Watered, two groups of n = 5); or transported, no water (Transported, two groups of n = 5). None of the horses had access to feed while on treatment. A commercial, single-deck, open top, 15.8-m-long trailer was divided into four compartments to accommodate the two Transported/Watered and two Transported groups at 1.77 m2 per horse. At 8, 17, 22, 27, 30, and 33 h after initiation of transport, the truck returned and stopped for 1 h to allow for data collection and to give the Transported/Watered and Penned/Watered horses 10 min of access to water in individual buckets. Treatments for the non-watered horses (Penned and Transported) were terminated after 30 h due to dehydration and fatigue, whereas the watered horses (Penned/Watered and Transported/Watered) could continue for another 2 h. Mean weight loss after 30 h was greater in the Penned (57.1 kg, 12.8%) and Transported (52.2 kg, 10.3%) groups than in the Transported/Watered (20.7 kg, 4.0%) and Penned/Watered (17 kg, 3.5%) groups (P < 0.0001). Respiration, heart rate, sodium, chloride, total protein, and osmolality were significantly elevated in the non-watered horses (P < 0.0001), and sodium, chloride, total protein, and osmolality greatly exceeded normal reference ranges, indicating severe dehydration. Although not statistically significant, the horses penned in full sun, with or without water, had a dehydration response that was slightly greater than that of the transported horses. Plasma cortisol concentrations had a significant time x treatment interaction (P < 0.0001), in which the Penned/Watered and Transported/Watered horses remained relatively consistent, whereas the Transported, and especially the Penned, horses' plasma cortisol concentrations greatly increased. Transporting healthy horses for more than 24 h during hot weather and without water will cause severe dehydration; transport for more than 28 h even with periodic access to water will likely be harmful due to increasing fatigue. PMID- 11048923 TI - Serum concentrations of IGF-I, estradiol-17beta, testosterone, and relative amounts of IGF binding proteins (IGFBP) in growing boars, barrows, and gilts. AB - The purpose of this research was to determine whether serum concentrations of steroids, IGF-I, and relative amounts of serum IGF-binding proteins (IGFBP) differ in growing boars (n = 11), barrows (n = 11), and gilts (n = 12) from 70 to 140 d of age. Pigs of similar age and weight were housed in pens of three or four and given ad libitum access to a 17% CP corn-soy diet and water. Pigs were weighed and blood samples were collected every 14 d beginning at 70 d of age. Serum concentrations of IGF-I and steroids were determined by RIA and relative amounts of IGFBP were determined by ligand blot analysis. By 84 d of age and continuing through 140 d of age, mean serum concentrations of IGF-I were greater (P < .05) in boars than in barrows or gilts. Relative amounts of 46-kDa IGFBP3 and 28-kDa IGFBP-4 were similar (P > .05) among pigs at 70 d of age; however, boars and barrows had greater (P < .05) relative amounts of 24-kDa IGFBP-4 and 41 kDa IGFBP-3 than gilts. Relative amounts of IGFBP-2 were greater (P < .01) in barrows than in gilts or boars at 70 d of age. From 84 d of age through 140 d of age, relative amounts of both forms of IGFBP-3 and the 28-kDa IGFBP-4 were greater (P < .05) in boars than in gilts or barrows. Relative amounts of IGFBP2 were greater (P < .05) in barrows than in gilts or boars at 98 d of age, but by 140 d of age relative amounts were greater (P < .05) in boars and barrows than in gilts. Mean serum concentrations of estradiol-17beta were similar (P > .05) between gilts and boars at 70 d of age, but by 98 d of age, and continuing through 140 d of age, mean serum concentrations of estradiol-17beta were greater (P < .05) in boars than in gilts. Mean serum concentrations of testosterone in boars increased (P < .05) with increasing age and were greatest at 128 and 140 d of age. Serum concentrations of testosterone were negatively correlated (P < .01) with relative amounts of serum IGFBP-2 but positively correlated (P < .01) with serum concentrations of IGF-I and estradiol-17beta. Serum concentrations of estradiol-17beta were positively correlated (P < .01) with serum concentrations of IGF-I in boars. Changes in serum concentrations of IGF-I and relative amounts of IGFBP resulting from changes in serum concentrations of estradiol-17beta and testosterone may contribute to growth differences observed among sexes. PMID- 11048924 TI - Bovine CAPN1 maps to a region of BTA29 containing a quantitative trait locus for meat tenderness. AB - Micromolar calcium activated neural protease (CAPN1) was investigated as a potential candidate gene for a quantitative trait locus (QTL) on BTA29 affecting meat tenderness. A 2,948-bp bovine cDNA containing the entire coding region of the gene was obtained, showing 91% identity to human CAPN1. The 716 AA protein predicted from this sequence shows 97% similarity (95% identity) to the 714 AA human protein. Analysis of the gene structure revealed that CAPN1 mRNA is encoded by at least 19 exons, and 11,055 bp of the gene were sequenced, including 17 introns. Two single nucleotide polymorphisms (SNP) were detected in intron 12 and were used to map bovine CAPN1 to the telomeric end of the BTA29 linkage group. This approximately coincides with the position of the QTL, demonstrating that CAPN1 protease is a positional candidate gene potentially affecting variation in meat tenderness in a bovine resource mapping population. PMID- 11048925 TI - Using measurements of muscle color, pH, and electrical impedance to augment the current USDA beef quality grading standards and improve the accuracy and precision of sorting carcasses into palatability groups. AB - This research was conducted to determine whether objective measures of muscle color, muscle pH, and(or) electrical impedance are useful in segregating palatable beef from unpalatable beef, and to determine whether the current USDA quality grading standards for beef carcasses could be revised to improve their effectiveness at distinguishing palatable from unpalatable beef. One hundred beef carcasses were selected from packing plants in Texas, Illinois, and Ohio to represent the full range of muscle color observed in the U.S. beef carcass population. Steaks from these 100 carcasses were used to determine shear force on eight cooked beef muscles and taste panel ratings on three cooked beef muscles. It was discovered that the darkest-colored 20 to 25% of the beef carcasses sampled were less palatable and considerably less consistent than the other 75 to 80% sampled. Marbling score, by itself, explained 12% of the variation in beef palatability; hump height, by itself, explained 8% of the variation in beef palatability; measures of muscle color or pH, by themselves, explained 15 to 23% of the variation in beef palatability. When combined together, marbling score, hump height, and some measure of muscle color or pH explained 36 to 46% of the variation in beef palatability. Alternative quality grading systems were proposed to improve the accuracy and precision of sorting carcasses into palatability groups. The two proposed grading systems decreased palatability variation by 29% and 39%, respectively, within the Choice grade and decreased palatability variation by 37% and 12%, respectively, within the Select grade, when compared with current USDA standards. The percentage of unpalatable Choice carcasses was reduced from 14% under the current USDA grading standards to 4% and 1%, respectively, for the two proposed systems. The percentage of unpalatable Select carcasses was reduced from 36% under the current USDA standards to 7% and 29%, respectively, for the proposed systems. These grading systems, which included requirements for maturity, marbling, hump height, and colorimeter readings, could be implemented into the current USDA beef quality grading standards and improve the accuracy and precision of sorting beef carcasses into palatability groups. At the least, measurements of muscle color or pH could be used in a branded-beef program to increase the palatability consistency of its beef products. PMID- 11048926 TI - Quantification of pork belly and boston butt quality attribute preferences of South Korean customers. AB - U.S. packers must have quantitative criteria for selection of pork bellies and Boston butts for export to South Korea. Pork bellies (IMPS 409A) and Boston butts (IMPS 409A, 406B, 407) were selected from normal production in a U.S. pork packing plant and transported to Seoul, South Korea, via seafreighter in refrigerated containers (frozen < -5 degrees C) or via air freight (fresh > 0 degrees C; frozen < -5 degrees C). Participants at the Seoul Food Show were surveyed about their preferences for specific quality attributes of these cuts. Bellies were selected to differ in seam fat content (low = < 20%, moderate = 20 to 40%, high = > 40% extractable fat), lean color (pale = L* > 50, medium = L* 48 50, dark = L* < 48), weight (3.36, 4.04, or 5.36 kg), state of refrigeration and packaging (frozen, poly-wrapped; chilled, poly-wrapped; frozen, vacuum-packaged; chilled, vacuum-packaged), shape (round, wavy, square), and belly thickness (3.81, 4.32, or 4.90 cm). Boston butts were selected to differ in USDA marbling score (Slight, Small, Modest, Moderate, and Slightly Abundant), seam fat content (low = < 10%, moderate = 10 to 20% fat, high = > 20% extractable fat), lean color (pale = L* > 44, medium = L* 40-42, dark = L* < 38), weight (2.91, 3.82, or 4.66 kg), state of refrigeration and packaging (frozen, poly-wrapped; chilled, poly wrapped; frozen, vacuum-packaged; chilled, vacuum-packaged), and shape (square, oblong, round). In Seoul, pork subprimals were tempered (if frozen), sliced, and arrayed by quality attribute and category in a retail display case. Over 4 d of testing, attendees (n = 210) of the food show were asked to rate the displayed samples for each quality attribute on a standardized ballot. Mid-weight (3.82 kg) Boston butts that displayed Moderate or higher USDA marbling scores with moderate amounts of seam fat, Japanese lean color scores of 2 or 4, round geometric shape, and that were vacuum-packaged and transported to Korea in the freshly chilled state best characterized the quality attribute preferences of respondents. Pork bellies that exhibited moderate amounts of seam fat, Japanese lean color scores of 3, square shape, belly thickness of 3.94 cm, approximate weight of 4.04 kg, and that were vacuum-packaged and transported to Korea in the freshly chilled state best met the quality needs of South Korean customers. PMID- 11048927 TI - The use of vitamin D3 to improve beef tenderness. AB - An experiment was designed to test the hypothesis that short-term oral administration of dietary vitamin D3 to beef cattle before slaughter would increase beef tenderness through greater calcium-activated calpain activity in postmortem aged skeletal muscle. Thirty continental crossbred steers were allotted randomly to three treatment groups housed in one pen. One group served as a control; two other groups were administered boluses with either 5 x 10(6) or 7.5 x 10(6) IU of vitamin D3 daily for 9 d. Cattle were slaughtered 1 d later. The longissimus lumborum was excised from each carcass 72 h postmortem and steaks removed at 3, 7, 14, and 21 d postmortem. The semimembranosus muscle (top round) was excised from each carcass 72 h postmortem and steaks removed at 7, 14, and 21 d postmortem. Blood plasma calcium concentration of cattle treated with 5 or 7.5 x 10(6) IU of vitamin D3 was higher (P < .05) than that of controls. Strip loin and top loin steaks from cattle fed supplemental doses of vitamin D3 had lower (P < .05) Warner-Bratzler (W-B) shear values at 14 d postmortem but were not significantly different from controls at 3, 7, or 21 d (strip loins) or 7 or 21 d (top rounds). No significant difference in strip loin steak tenderness was observed by sensory panel at 14 d postmortem (P < .17) between steaks from control and vitamin D3-treated steers. At 14 d postmortem, strip loin and top round steaks from cattle fed 5 x 10(6) IU of vitamin D3, but not from those given 7.5 x 10(6) IU, showed more proteolysis (P < .05) than did steaks from control cattle, based on Western blotting analysis. Therefore, the use of supplemental dietary vitamin D3 given daily for 9 d before slaughter did improve tenderness (lower W-B shear values) of 14-d postmortem aged beef. Increased proteolysis seems to be the mechanism of tenderization. PMID- 11048928 TI - The effects of calcium benzoate in diets with or without organic acids on dietary buffering capacity, apparent digestibility, retention of nutrients, and manure characteristics in swine. AB - Eight barrows (Yorkshire x [Finnish Landrace x Dutch Landrace]), initially 30 kg BW, were fitted with ileal cannulas to evaluate the effects of supplementing Ca benzoate (2.4%) and organic acids (OA) in the amount of 300 mEq acid/kg feed on dietary buffering capacity (BC), apparent digestibility and retention of nutrients, and manure characteristics. Swine were allotted in a 2 x 4 factorial arrangement of treatments according to a cyclic (8 x 5) changeover design. Two tapioca-corn-soybean meal-based diets were formulated without and with acidogenic Ca benzoate. Each diet was fed in combination with OA (none, formic, fumaric, or n-butyric acid). Daily rations were equal to 2.8 x maintenance requirement (418 kJ ME/BW(.75)) and were given in two portions. Chromic oxide (.25 g/kg) was used as a marker. On average, Ca benzoate lowered BC by 54 mEq/kg feed. This salt enhanced (P < .05) the ileal digestibility (ID) of DM, OM, arginine, isoleucine, leucine, phenylalanine, alanine, aspartic acid, and tyrosine (by up to 2.4 percentage units). Also, the total tract digestibility (TD) of DM, ash, Ca and GE, and Ca retention (percentage of intake) was greater (P < .05) in swine fed Ca benzoate, whereas N retention remained unaffected. Addition of all OA (formic and n-butyric acid, in particular) exerted a positive effect (P < .05) on the ID of amino acids (except for arginine, methionine, and cysteine). A similar effect (P < .05) was found for the TD of DM, OM, CP, Ca and total P and for the retention of N and Ca. In swine fed Ca benzoate, urinary pH decreased by 1.6 units (P < .001). In conclusion, dietary OA have a beneficial effect on the apparent ileal/total tract nutrient digestibilities, and Ca benzoate increased urine acidity, which could be effective against a rapid ammonia emission from manure of swine. PMID- 11048929 TI - The effect of texturized vegetable protein containing soy carbohydrate on oroileal transit of chromic oxide in cannulated dogs. AB - Texturized vegetable protein (TVP) from soy is widely used in dog foods but contains indigestible carbohydrate, which may affect intestinal transit. This study was conducted to determine whether TVP affects oroileal transit of the marker chromic oxide (Cr2O3). Four canned diets with Cr2O3 added were fed to eight cannulated mixed-breed dogs in a Latin-square design. The four diets contained reciprocal proportions of protein from TVP (0 to 57%) and from beef (100 to 43%). Ileal effluent was collected during wk 3 of each diet period, prececal apparent digestibility was measured on d 1 to 3 (eight dogs), and rate of appearance of Cr2O3 and chyme was measured on d 4 (six dogs). There was a postprandial delay before any chyme or Cr2O3 was collected, but from 2 to 8 h postprandially the rate of excretion was almost constant (approximately 11%/h). As TVP increased, mean prececal digestibility of protein and carbohydrate decreased from 77 to 71% and from 80 to 62%, respectively. Chyme excretion from 2 to 8 h postprandially increased from 5.8 to 8.3 g DM/h (P < .0001) as TVP increased but times for transit of Cr2O3 and appearance of chyme (as percentage total collected/hour) did not change. Mean times to first appearance and 15, 50, and 95% excretion were 76, 160, 347, and 707 min for Cr2O3 and 60, 147, 338, and 712 min for chyme, respectively. Mean Cr2O3 recovery was 93%. In conclusion, small intestinal transit of Cr2O3 was unaffected by increasing dietary TVP despite marked changes in carbohydrate digestibility. PMID- 11048930 TI - Interrelationships between dietary lysine, sex, and porcine somatotropin administration on growth performance and protein deposition in pigs between 80 and 120 kg live weight. AB - Sixty male and 60 female crossbred pigs were allocated to an experiment to investigate the effects of porcine somatotropin (pST) administration (0 or 6 mg/d) and dietary lysine content on growth performance, tissue deposition, and carcass characteristics over the live weight range of 80 to 120 kg. Pigs receiving pST were given diets containing 6.9, 7.8, 8.8, 9.7, 10.6, or 11.5 g lysine/kg, whereas control pigs received diets containing 4.8, 5.8, 6.9, 7.8, 8.8 or 9.7 g lysine/kg. These dietary levels ranged from 0.40 to 0.70 g available lysine/MJ of DE for pST-treated pigs and from 0.28 to 0.58 g available lysine/MJ of DE for control pigs. Pigs were individually housed in pens, and there were five replicates of each treatment. All diets contained 14.5 MJ of DE/kg and were offered for ad libitum consumption to pigs between 80 and 120 kg live weight. Growth rate increased exponentially and food conversion ratio (FCR) decreased exponentially with increasing levels of lysine. In addition, there was a significant sex x pST interaction such that pST reduced the sex difference in FCR. Growth rate was faster in boars than in gilts and was increased by pST at the higher levels of dietary lysine. Similarly, FCR was lower for boars than for gilts and was decreased by pST at the higher dietary lysine levels. The optimum growth rate and FCR were defined as the lysine level at which growth rate and FCR were 95% and 105%, respectively, of the lysine plateau. The optimum growth rate and FCR were achieved at similar dietary lysine contents and were approximately 0.35 and 0.52 g available lysine/MJ of DE for control and pST-treated pigs, respectively. Protein deposition in the carcass increased exponentially with increasing dietary lysine level, was higher in boars than in gilts, and was increased by pST at the higher dietary lysine contents. Sex had no effect on dietary lysine required to maximize protein deposition. The dietary lysine contents required to ensure 95% of plateau protein deposition of 104 and 153 g/d were 0.39 and 0.55 g available lysine/MJ of DE for control and pST-treated pigs, respectively. The increase in lysine requirement with pST seems to be commensurate with the increase in protein deposition. PMID- 11048931 TI - Variability among sources and laboratories in analyses of wheat middlings. NCR-42 Committee on Swine Nutrition. AB - A cooperative research study was conducted by members of a regional committee (North Central Regional Committee on Swine Nutrition [NCR-42]) to assess the variability in nutrient composition (DM, CP, Ca, P, Se, NDF, and amino acids) of 14 sources of wheat middlings from 13 states (mostly in the Midwest). A second objective was to assess the analytical variability in nutrient assays among 20 laboratories (labs; 14 experiment station labs and six commercial labs). Wheat middlings were obtained from each participating station's feed mill. The bulk density of the middlings ranged from 289 to 365 g/L. The number of labs that analyzed samples were as follows: DM and CP, 20; Ca, 16; P, 15; Se, 7; NDF, 10; and amino acids, 9. Each lab used its own analytical procedures. The middlings averaged 89.6% DM, 16.2% CP, .12% Ca, .97% P, 36.9% NDF, .53 mg/kg Se, .66% lysine, .19% tryptophan, .54% threonine, .25% methionine, .34% cystine, .50% isoleucine, and .73% valine. As expected, there was considerable variation in nutrient composition among the 14 sources (P < .01), especially for Ca (.08 to .30%) and Se (.05 to 1.07 mg/kg). "Heavy" middlings (high bulk density, >335 g/L), having a greater proportion of flour attached to the bran, were lower in CP, lysine, P, and NDF than "light" middlings (<310 g/L), having cleaner bran, resulting in negative correlations between bulk density and CP (r = -.61), lysine (r = -.59), P (r = -.54), and NDF (r = -.81). Each 1-percentage-point increase in CP in the wheat middlings was associated with .0235 (r2 = .61) and 2.1 (r2 = .39) percentage-point increases in lysine and NDF, respectively. Lysine content was associated with NDF, CP, and bulk density of wheat middlings (r2 = .88). There was considerable variation among laboratories (P < .01) in analysis of all nutrients. The CV among sources (100 x sigmaS/mean) was greater than among labs (100 x sigmaL/mean) for CP, Ca, P, Se, and NDF, but the CV among labs was greater than that among sources for DM and all of the amino acids except lysine and phenylalanine. PMID- 11048932 TI - Nitrogen metabolism and fertility in cattle: I. Adaptive changes in intake and metabolism to diets differing in their rate of energy and nitrogen release in the rumen. AB - The ruminal degradability, intake, and metabolism of diets differing in their relative rate of energy and nitrogen release in the rumen were characterized prior to their use in a study of the effects of high peripheral levels of ammonia on reproductive function in cattle. In a 2 x 2 factorial experiment, replicated four times, 16 heifers were offered isocaloric and isonitrogenous diets containing two sources of fermentable carbohydrate, fiber (slow energy release, SE) or starch (fast energy release, FE), and two rates of nitrogen release, which were either synchronous (S) or asynchronous (A) to that of energy release. Throughout the experiment, the amount of feed offered was held constant, at a level equivalent to 1.5 x maintenance. Four ruminally fistulated sheep were used to determine the in situ degradability of these diets. The 16 heifers were bled before feeding at 0800 and at 0900, 1000, 1100, 1200, 1400, and 1600 on d 0 (introduction to dietary treatments) and on d 4, 7, 11, 14, 21, and 28. Diet refusals were recorded at hourly intervals after feeding. The rapidly degradable nitrogen fraction of the SE:A and FE:A diets was greater than that of the SE:S and FE:S diets. Postprandial jugular plasma ammonia levels rose to a peak of around 300 micromol/L in heifers offered the SE:A and FE:A diets but did not rise in heifers offered the SE:S and FE:S diets. All feed offered was consumed within 1 h on diets SE:S and FE:S throughout the experiment. The proportion of feed consumed within 1 h of feeding declined from 100% on d 0 to around 70 and 56% by d 21 for heifers given the SE:A and FE:A diets, respectively. Peak postprandial plasma ammonia levels were accordingly lower, at around 160 micromol/L. Plasma urea levels averaged 7 mmol/L and were unaffected by dietary treatment. High plasma ammonia levels were associated with a suppression in the normal postprandial rise in insulin. There was no significant metabolic adaptation to high-ammonia-generating diets, and heifers given these diets modified their pattern of intake in an apparent attempt to avoid excessively high levels of plasma ammonia. PMID- 11048933 TI - Nitrogen metabolism and fertility in cattle: II. Development of oocytes recovered from heifers offered diets differing in their rate of nitrogen release in the rumen. AB - In vitro blastocyst production was determined for oocytes recovered postmortem from 48 beef x dairy heifers offered low (Low NH3) or high (High NH3) plasma ammonia-generating diets during the period of late antral follicle development. Following the establishment of a reference estrus (d 0), the experimental diets were offered for an 18-d period starting on d 3 and during which a second estrus was induced (d 16) 4 d before the animals were slaughtered. Blood samples collected at varying intervals were analyzed for ammonia, urea, progesterone, and LH. Ovarian folliculogenesis was monitored daily by transrectal ultrasonography. Ovaries were collected at slaughter and cumulus-oocyte complexes were aspirated from small (1 to 4 mm) and medium-sized (> 4 to 8 mm) sized follicles. In vitro matured and -fertilized putative d-1 zygotes were cultured for a further 7 d in vitro and embryo development and metabolism were assessed. Relative to the low NH3-generating diet, the high-NH3-generating diet increased peak postprandial levels of plasma ammonia (326.1 +/- 43.3 vs 52.1 +/- 7.4 micromol/L; P < .001), mean levels of plasma urea (7.0 vs 5.7 mmol/L; SED = .2; P < .001), peak levels of plasma progesterone prior to induced luteolysis (8.9 +/- .4 vs 6.8 +/- .3 microg/L; P < .001), and follicular fluid levels of ammonia (267 +/- 18 vs 205 +/ 20 nmol/mL; P < .05) and progesterone (351 +/- 69 vs 199 +/- 26 ng/mL; P < .05). The timing and level of the preovulatory LH surge was not affected by dietary treatment. Of oocytes cultured, cleavage (47.4 vs 62.4%; P = .02) and blastocyst production (10.9 vs 20.6%; P = .06) rates were reduced when the oocytes were derived from heifers offered the high- rather than the low-NH3-generating diets. There were interactions between dietary treatment and follicle size class, which indicated that fewer blastocysts were produced from cleaved oocytes derived from medium-sized follicles of heifers offered the high-NH3 treatment but that de novo protein synthesis was increased in such embryos. In conclusion, exposure to high levels of ammonia and(or) urea in vivo can significantly compromise the subsequent capacity of oocytes to develop to blastocysts in vitro, and oocytes recovered from medium-sized follicles are particularly sensitive to this effect. PMID- 11048934 TI - Nutritional regulation of the genes encoding the acid-labile subunit and other components of the circulating insulin-like growth factor system in the sheep. AB - In sheep, perinatal maturation of the endocrine arm of the insulin-like growth factor (IGF) system is characterized by two developmental events. First, concentrations of circulating IGF-I increase rapidly after birth and become responsive to changes in nutrition and growth hormone (GH). Second, the liver initiates synthesis of a serum protein called the acidlabile subunit (ALS). The acid-labile subunit promotes the endocrine actions of IGF-I and -II by recruiting them to long-lived complexes of 150 kDa. In this study, we examined the effect of nutrition on hepatic expression of the ALS gene around the time of birth and later in life. Expression of genes encoding other components of the circulating IGF system was also measured. At d 130 of fetal life, fetuses suffering from chronic undernutrition caused by placental insufficiency had lower expression of the ALS and IGF-I genes than well-nourished fetuses, but they did not have any changes in the expression of the IGF-binding protein (IGFBP)-2 or IGFBP-3 genes. In early postnatal life, hepatic gene expression was analyzed between d 12 and 38 in lambs fed a milk replacer at levels sustaining weight gains of 150 or 337 g/d. The lower plane of nutrition decreased the expression of the ALS, IGF-I, and GH receptor genes and increased the expression of the IGFBP-2 gene; expression of the IGFBP-3 gene was not affected by nutrition at this stage of life. Finally, hepatic gene expression was measured in 3-mo-old lambs offered ad libitum levels of a balanced diet or of a diet limiting for both energy and protein. Although the rate of growth of the lambs fed the limiting diet was reduced by 38%, the only effect detected in hepatic gene expression was a ninefold increase in the abundance of IGFBP-2 mRNA. Overall, these results indicate that undernutrition during late fetal and early postnatal life delays hepatic expression of the ALS gene and final maturation of the endocrine IGF system. PMID- 11048935 TI - Quantitative relationship of systemic virus concentration on growth and immune response in pigs. AB - Ninety-six pigs from a herd naive for porcine reproductive and respiratory syndrome (PRRS) virus were weaned (10 +/- 3 d of age), penned individually in isolation rooms, and, at 29 +/- 4 d of age, oronasally inoculated with a 2-mL dose of 10(4.3) JA142 PRRS virus/ mL. Body weight; feed intake; and serum concentrations of PRRS virus, interferon, and alpha1-acylglycoprotein were determined for each pig every 4 d on d -8 to 24 postinoculation to quantify the effect of PRRS exposure on the immune response and growth of pigs. Another objective was to determine whether a quantitative relationship between a measure of systemic (serum) virus concentration and pig growth exists. Serum PRRS virus and interferon peaked at 10(5) virus/mL and 69% protection, respectively, at 4 d postinoculation and then declined steadily. Serum alpha1-acylglycoprotein concentration peaked at 12 d postinoculation. Pig weight gains and feed intake were reduced sharply in the initial 8 d postinoculation and to a lesser degree for 24 d postinoculation. The serum concentration of virus and to a lesser degree serum concentrations of interferon and alpha1-acylglycoprotein were quantitatively related to body weight gain and feed intake. The magnitude of the relationship was dependent on the stage of recovery from PRRS infection. Specifically, each log increase in serum virus concentration was associated with a reduction of 4-d pig gain and feed intake of .047 kg and .189 kg, respectively, in 5.5-kg pigs 4 d postinoculation and .085 kg and .036 kg, respectively, in 12.5 kg pigs at 20 d postinoculation. Based on these data, factors that minimize the systemic presence of a virus in pigs result in improvements in pig growth that are quantitatively related to the degree of systemic virus elimination or minimization. PMID- 11048936 TI - Energy metabolism in lactating beef heifers. AB - To obtain measurements of energy balance in lactating beef cows, respiration calorimetry and digestion trials were conducted using seven lactating (613 kg BW) and three nonlactating (598 kg BW) Hereford x Angus heifers fed a pelleted 75% alfalfa:25% concentrate diet. Five measurements of energy balance were obtained at 6- to 7-wk intervals beginning 6 to 10 wk postpartum in lactating heifers and at 6-wk intervals in nonlactating heifers. Milk yield was measured using a combination of weigh-suckle-weigh and machine milking to adapt heifers to milking by machine without the use of oxytocin. Heifers were milked only by machine during measurements of energy balance. Weekly milk yield averages ranged from 8.2 kg/d at wk 5 postpartum to 3.2 kg/d at wk 32 postpartum. When scaled to BW(.75), the regression of NE1 on ME intake and the regression of ME intake on NE1 were remarkably similar to previously published regressions for measurements obtained from lactating Holstein-Friesian cows. The average daily maintenance energy requirement from these regressions was 503 kJ ME/kg BW(.75), a value similar to the average value reported previously for lactating Holstein-Friesian cows (488 kJ/kg (BW.75)). This is in contrast to numerous published comparisons of the maintenance requirements of cattle breed types in the nonlactating state and current NRC standards for estimating maintenance energy requirements of beef and dairy cattle. The results of the present study suggest that when expressed on the basis of BW(.75) the efficiency of utilization of incremental ME above maintenance for milk and tissue energy (i.e., NE1) is similar among lactating Hereford x Angus heifers and lactating Holstein-Friesian cows. The breeds differ in terms of their propensity for milk yield and the resulting partition of ME between milk synthesis and tissue energy retention. PMID- 11048937 TI - A kinetic model of phosphorus metabolism in growing goats. AB - The effect of increasing phosphorus (P) intake on P utilization was investigated in balance experiments using 12 Saanen goats, 4 to 5 mo of age and weighing 20 to 30 kg. The goats were given similar diets with various concentrations of P, and 32P was injected to trace the movement of P in the body. A P metabolism model with four pools was developed to compute P exchanges in the system. The results showed that P absorption, bone resorption, and excretion of urinary P and endogenous and fecal P all play a part in the homeostatic control of P. Endogenous fecal output was positively correlated to P intake (P < .01). Bone resorption of P was not influenced by intake of P, and P recycling from tissues to the blood pool was lesser for low P intake. Endogenous P loss occurred even in animals fed an inadequate P diet, resulting in a negative P balance. The extrapolated minimum endogenous loss in feces was .067 g of P/d. The minimum P intake for maintenance in Saanen goats was calculated to be .61 g of P/d or .055 g of P/(kg(.75) x d) at 25 kg BW. Model outputs indicate greater P flow from the blood pool to the gut and vice versa as P intake increased. Intake of P did not significantly affect P flow from bone and soft tissue to blood. The kinetic model and regressions could be used to estimate P requirement and the fate of P in goats and could also be extrapolated to both sheep and cattle. PMID- 11048938 TI - Effects of soybean oil and dietary copper on ruminal and tissue lipid metabolism in finishing steers. AB - An experiment was conducted to determine the effects of Cu and soybean oil (SBO) supplementation on ruminal and tissue lipid metabolism and carcass characteristics in finishing steers. Sixty Angus steers (369.0 +/- 10.1 kg) were stratified by weight and randomly assigned to treatments in a 2 x 2 factorial arrangement, with factors being 0 or 20 mg of supplemental Cu/kg DM from Cu sulfate and 0 or 4% SBO. Steers were fed a high-concentrate basal diet that contained 5.3 mg Cu/kg DM. Average daily gain and feed intake were reduced (P < 0.01) by SBO but were not affected by Cu. Gain:feed ratio was not affected by treatment. Liver Cu concentrations were higher (P < 0.01) in steers receiving supplemental Cu and lower (P < 0.04) in SBO-supplemented steers. Copper supplementation tended to reduce (P < 0.12) and SBO supplementation tended to increase (P < 0.11) serum cholesterol concentrations. Backfat depth was reduced (P < 0.10) by Cu and SBO supplementation. Marbling scores and longissimus muscle lipid content were not affected by Cu supplementation; however, SBO supplementation reduced (P < 0.01) marbling scores. Longissimus muscle polyunsaturated fatty acids tended to be increased (P < 0.14) in Cu-supplemented steers. Longissimus muscle C18-conjugated dienes and the 18:1 trans isomer were increased (P < 0.05) in SBO-supplemented steers. Ruminal fluid 18:3 was increased (P < 0.05) and the 18:1 trans isomer was decreased (P < 0.05) in Cu-supplemented steers. These results indicate that as little as 20 mg of supplemental Cu/kg DM can reduce backfat and may alter lipid metabolism in steers fed high-concentrate diets. PMID- 11048939 TI - Methionine imbalance and toxicity in calves. AB - The occurrence of methionine imbalance and toxicity was examined using 70- and 100-kg Holstein bull calves. The animals had been trained to maintain reflex closure of the reticular groove after weaning at 5 wk of age, and Trials 1 (n = 30) and 2 (n = 24) were conducted on animals at 7 and 12 wk of age, respectively. Calves received a corn-soybean meal diet in Trial 1 and a corn-corn gluten meal diet in Trial 2. In Trial 1, postruminal administration of 6 g of DL-methionine/d increased ADG, feed intake, gain/feed, and N retention compared with a control group receiving N-free supplement. However, the administration of 12 g of DL methionine/d did not improve these variables, whereas both 18 and 24 g/d resulted in BW loss and decreased gain/feed and N utilization efficiency. In Trial 2, postruminal administration of 16 g/d of L-lysine from L-lysine monohydrochloride increased ADG, gain/feed, and N utilization efficiency compared with a control group receiving a N-free supplement. The administration of 8 g of DL-methionine/d in addition to L-lysine did not exert an adverse effect on these variables. However, the additional supplementation of 16 and 24 g of DLmethionine/d negated the improvement, whereas 32 g/d resulted in BW loss and decreased gain/feed and N utilization efficiency. These results showed that a methionine imbalance and toxicity occurred in calves with even a modest excess of DL-methionine, and 70-kg calves were more susceptible to methionine toxicity than 100-kg calves. Plasma concentrations of branched-chain amino acids and phenylalanine linearly decreased with increasing amounts of additional DL-methionine from 0 to 32 g/d in Trial 2. However, such a decrease occurred mainly within the range from 0 to 12 g/d in Trial 1. This decrease was suggested to occur in relation to methionine metabolism via the transsulfuration pathway. PMID- 11048940 TI - Evaluation of ewe and lamb immune response when ewes were supplemented with vitamin E. AB - Fifty-two Targhee twin-bearing ewes were used in a factorial arrangement of treatments to investigate the role of supplemental vitamin E (vit E); 0 (NE) vs 400 IU of vit E x ewe x (-1)d(-1) (E) and parainfluenza type 3 (PI3) vaccination; none (NP) vs PI3 vaccination (P) in immune function. Parainfluenza type 3 vaccination was used to evoke an immune response. Ewes receiving PI3 were vaccinated at 49 and 21 d before the expected lambing date. Ewes receiving vit E were orally dosed daily, 32 to 0 d before lambing. Blood was collected from ewes at the time of the initial PI3 vaccination and 4 h postpartum. Blood was collected from lambs (n = 104) at 3 d postpartum. Ewe and lamb sera were analyzed for anti-PI3 antibody titers, immunoglobulin G (IgG) titers, and vit E concentrations. Colostrum was collected 4 h postpartum and analyzed for IgG. The model for ewe and lamb analysis included the main effects of vit E and PI3, sex (lambs model only), and their interactions. No interactions were detected (P > 0.20) for any ewe or lamb variables. Serum anti-PI3 titers were greater (P < 0.01) in P ewes and their lambs than NP ewes and their lambs. Serum vit E concentrations were greater (P < 0.01) in E ewes and their lambs than NE ewes and their lambs. Colostral IgG titers and serum anti-PI3 titers did not differ (P > 0.20) between E and NE ewes. Serum IgG titers in E ewes and their lambs did not differ (P > 0.15) from IgG titers in NE ewes and their lambs. Lamb anti-PI3 titers did not differ (P = 0.76) between lambs reared by E and NE ewes. These results indicate that, although supplemental vit E to the ewe increased lamb serum vit E concentration, it had no effect on measures used in this study to assess humoral immunity in the ewe or passive immunity to the lamb. PMID- 11048941 TI - Dietary copper effects on lipid metabolism and circulating catecholamine concentrations in finishing steers. AB - Forty-eight Angus and Hereford x Angus steers were used to determine the effects of copper (Cu) on lipid and catecholamine metabolism. Steers were stratified by weight within breed and randomly assigned to treatments. Treatments consisted of 0 (control, no supplemental Cu), 10, or 40 mg of supplemental Cu (from Cu2(OH)3Cl)/kg DM. Steers were fed a corn silage-soybean meal-based growing diet for 42 d. Animals were then switched to a high-concentrate finishing diet and remained on the same dietary treatments. On d 70, indwelling jugular catheters were nonsurgically inserted into five steers per treatment. Blood samples were obtained from steers after a 24-h period of feed withdrawal, 1 h after feeding, and after i.v. administration of norepinephrine and were subsequently analyzed for nonesterified fatty acid (NEFA) and catecholamine concentrations. Average daily gain over the finishing period was higher (P < 0.06) in steers receiving supplemental Cu. Serum total cholesterol concentrations were reduced (P < 0.05) on d 84 and 112 in steers supplemented with Cu. Serum norepinephrine (P < 0.14) and NEFA concentrations following feed withdrawal tended (P < 0.12) to be higher in Cu-supplemented steers. Postfeeding norepinephrine concentrations tended to be higher (P < 0.14) in Cu-supplemented steers. Nonesterifled fatty acid concentrations were lower (P < 0.10) in Cu-supplemented steers after norepinephrine administration. Backfat depth was decreased (P < 0.10) and longissimus muscle polyunsaturated fatty acid percentages were increased (P < 0.10) in steers receiving supplemental Cu. These results indicate that Cu addition to a finishing diet containing 5 mg Cu/kg DM alters lipid metabolism. The reduction in backfat depth may be due to copper altering catecholamine metabolism in steers. PMID- 11048942 TI - Site of administration and duration of feeding oleamide to cattle on feed intake and ruminal fatty acid concentrations. AB - Two experiments were conducted to determine effects of oleamide on feed intake and ruminal fatty acids when the oleamide was introduced in the feed vs through a ruminal fistula (Exp. 1) or the oleamide was fed for an extended (9-wk) length of time (Exp. 2). In Exp. 1, four nonlactating Holstein cows, each fitted with a ruminal cannula, were fed four diets in a 4 x 4 Latin square design. Each period lasted 2 wk. Diets consisted of 48% corn silage and 52% concentrate on a DM basis. One diet contained no added fat (control) and a second diet contained 4.2% oleic acid. The remaining two diets were designed to expose cows to 4.2% amide (as oleamide) either through the feed (AF) or by administering oleamide into the rumen (AR) each day through the ruminal cannula. The AF diet reduced DMI similarly to results reported previously for lactating dairy cows and sheep. Intake of the oleic acid diet was intermediate between the control and AF diets. Dry matter intake was reduced by AR similarly to the AF diet. The acetate:propionate ratio in samples of ruminal contents was reduced by oleic acid but not by AF or AR. In Exp. 2, 12 steers were divided into three equal groups of two Angus and two Simmental x Angus crosses, and each group was assigned a diet containing either no added fat (control), 4% oleamide, or 4% high-oleic canola oil. All steers had ad libitum access to feed and water. Dry matter intake by steers fed the canola oil diet was not different from that by steers fed the control diet when averaged over the first 3 wk, the last 3 wk, or over the entire 9-wk study. Oleamide reduced DMI 4 kg/d over the first 3 wk of the study. However, DMI of the oleamide diet consistently increased over the 9-wk study, resulting in wk 7 to 9 DMI that was not different from that of steers fed the control diet. These results show that the reduction in feed intake when oleamide is added to cattle rations can be attributed more to physiological responses than to an undesirable unique taste or odor of the oleamide. In finishing beef steers, the decreased intake induced by oleamide was most severe during the first 1 or 2 wk of feeding but gradually lessened over time until it nearly returned to normal by wk 9. PMID- 11048943 TI - Rapid communication: a single-strand conformation polymorphism in the ovine interleukin-2 (IL-2) gene. PMID- 11048944 TI - Rapid communication: cloning of bovine serum amyloid A3 cDNA. PMID- 11048945 TI - Anticalins versus antibodies: made-to-order binding proteins for small molecules. AB - Engineering proteins to bind small molecules presents a challenge as daunting as drug discovery, for both hinge upon our understanding of receptor-ligand molecular recognition. However, powerful techniques from combinatorial molecular biology can be used to rapidly select artificial receptors. While traditionally researchers have relied upon antibody technologies as a source of new binding proteins, the lipocalin scaffold has recently emerged as an adaptable receptor for small molecule binding. 'Anticalins', engineered lipocalin variants, offer some advantages over traditional antibody technology and illuminate features of molecular recognition between receptors and small molecule ligands. PMID- 11048946 TI - Eek, a XenoMouse: Abgenix, Inc. PMID- 11048947 TI - Cartilage degradation by stimulated human neutrophils: reactive oxygen species decrease markedly the activity of proteolytic enzymes. AB - BACKGROUND: Although neutrophilic granulocytes clearly contribute to cartilage degradation in rheumatic diseases, it is unclear if reactive oxygen species (ROS) or proteolytic enzymes are the most important components in cartilage degradation and how they interact. RESULTS: Neutrophils were stimulated by chemicals conferring a different degree of ROS formation and enzyme release. Supernatants of neutrophils were incubated with thin slices of pig articular cartilage. Supernatants of cartilage were assayed by NMR spectroscopy, MALDI-TOF mass spectrometry and relevant biochemical methods. Stimulation conditions of neutrophils correlated well with the extent of cartilage degradation. Due to the release of different enzymes, cartilage degradation could be best monitored by NMR since mainly low-mass degradation products were formed. Astonishingly, the suppression of the formation of ROS resulted in decreased cartilage degradation. CONCLUSION: ROS formed by neutrophils are not directly involved in cartilage degradation but influence the activity of proteolytic enzymes, which are the main effectors of cartilage degradation. PMID- 11048948 TI - Epoxide electrophiles as activity-dependent cysteine protease profiling and discovery tools. AB - BACKGROUND: Analysis of global changes in gene transcription and translation by systems-based genomics and proteomics approaches provides only indirect information about protein function. In many cases, enzymatic activity fails to correlate with transcription or translation levels. Therefore, a direct method for broadly determining activities of an entire class of enzymes on a genome-wide scale would be of great utility. RESULTS: We have engineered chemical probes that can be used to broadly track activity of cysteine proteases. The structure of the general cysteine protease inhibitor E-64 was used as a scaffold. Analogs were synthesized by varying the core peptide recognition portion while adding affinity tags (biotin and radio-iodine) at distal sites. The resulting probes containing a P2 leucine residue (DCG-03 and DCG-04) targeted the same broad set of cysteine proteases as E-64 and were used to profile these proteases during the progression of a normal skin cell to a carcinoma. A library of DCG-04 derivatives was constructed in which the leucine residue was replaced with all natural amino acids. This library was used to obtain inhibitor activity profiles for multiple protease targets in crude cellular extracts. Finally, the affinity tag of DCG-04 allowed purification of modified proteases and identification by mass spectrometry. CONCLUSIONS: We have created a simple and flexible method for functionally identifying cysteine proteases while simultaneously tracking their relative activity levels in crude protein mixtures. These probes were used to determine relative activities of multiple proteases throughout a defined model system for cancer progression. Furthermore, information obtained from libraries of affinity probes provides a rapid method for obtaining detailed functional information without the need for prior purification/identification of targets. PMID- 11048949 TI - Tuning chemotactic responses with synthetic multivalent ligands. AB - BACKGROUND: Multivalent ligands have been used previously to investigate the role of ligand valency and receptor clustering in eliciting biological responses. Studies of multivalent ligand function, however, typically have employed divalent ligands or ligands of undefined valency. How cells respond to multivalent ligands of distinct valencies, which can cluster a signaling receptor to different extents, has never been examined. The chemoreceptors, which mediate chemotactic responses in bacteria, are localized, and clustering has been proposed to play a role in their function. Using multivalent ligands directed at the chemoreceptors, we hypothesized that we could exploit ligand valency to control receptor occupation and clustering and, ultimately, the cellular response. RESULTS: To investigate the effects of ligand valency on the bacterial chemotactic response, we generated a series of linear multivalent arrays with distinct valencies by ring-opening metathesis polymerization. We report that these synthetic ligands elicit bacterial chemotaxis in both Escherichia coli and Bacillus subtilis. The chemotactic response depended on the valency of the ligand; the response of the bacteria can be altered by varying chemoattractant ligand valency. Significantly, these differences in chemotactic responses were related to the ability of the multivalent ligands to cluster chemoreceptors at the plasma membrane. CONCLUSIONS: Our results demonstrate that ligand valency can be used to tune the chemotactic responses of bacteria. This mode of regulation may arise from changes in receptor occupation or changes in receptor clustering or both. Our data implicate changes in receptor clustering as one important mechanism for altering cellular responses. Given the diverse events modulated by changes in the spatial proximity of cell surface receptors, our results suggest a general strategy for tuning biological responses. PMID- 11048951 TI - Virtually unidirectional binding of TBP to the AdMLP TATA box within the quaternary complex with TFIIA and TFIIB. AB - BACKGROUND: The TATA box binding protein (TBP) is required by all three RNA polymerases for the promoter-specific initiation of transcription. All eukaryotic TBP-DNA complexes observed in crystal structures show the conserved C-terminal domain of TBP (TBPc) bound to the TATA box in a single orientation that is consistent with assembly of a preinitiation complex (PIC) possessing a unique polarity. The binding of TBP to the TATA box is believed to orient the PIC correctly on the promoter and can function as the rate-limiting step in PIC assembly. Previous work performed with TBP from Saccharomyces cerevisiae (yTBP) showed that, despite the oriented binding of eukaryotic TBP observed in crystal structures, yTBP in solution does not orient itself uniquely on the adenovirus major late promoter (AdMLP) TATA box. Instead, yTBP binds the AdMLP as a mixture of two orientational isomers that are related by a 180 degree rotation about the pseudo-dyad axis of the complex. In addition, these orientational isomers are not restricted to the 8 bp TATA box, but rather bind a distribution of sites that partially overlap the TATA box. Two members of the PIC, general transcription factor (TF) IIB and TFIIA individually enhance the orientational and axial specificity of yTBP binding to the TATA box, but fail to fix yTBP in a single orientation or a unique position on the promoter. RESULTS: We used an affinity cleavage assay to explore the combined effects of TFIIA and TFIIB on the axial and orientational specificity of yTBP. Our results show that the combination of TFIIA and TFIIB affixes yTBP in virtually a single orientation as well as a unique location on the AdMLP TATA box. Ninety-five percent of the quaternary TBP TFIIA-TFIIB-TATA complex contained yTBP bound in the orientation expected on the basis of crystallographic and genetic experiments, and more than 70% is restricted axially to the 8 bp sequence TATAAAAG. CONCLUSIONS: Although yTBP itself binds to the TATA box without a high level of orientational or axial specificity, our data show that a small subset of general TFs are capable of uniquely orienting the PIC on the AdMLP. Our results, in combination with recent data concerning the pathway of PIC formation in yeast, suggest that transcription could be regulated during both early and late stages of PIC assembly by general factors (and the proteins to which they bind) that influence the position and orientation of TBP on the promoter. PMID- 11048950 TI - Chemical synthesis and biological properties of pyridine epothilones. AB - BACKGROUND: Numerous analogs of the antitumor agents epothilones A and B have been synthesized in search of better pharmacological profiles. Insights into the structure-activity relationships within the epothilone family are still needed and more potent and selective analogs of these compounds are in demand, both as biological tools and as chemotherapeutic agents, especially against drug resistant tumors. RESULTS: A series of pyridine epothilone B analogs were designed, synthesized and screened. The synthesized compounds exhibited varying degrees of tubulin polymerization and cytotoxicity properties against a number of human cancer cell lines depending on the location of the nitrogen atom and the methyl substituent within the pyridine nucleus. CONCLUSIONS: The biological screening results in this study established the importance of the nitrogen atom at the ortho position as well as the beneficial effect of a methyl substituent at the 4- or 5-position of the pyridine ring. Two pyridine epothilone B analogs (i.e. compounds 3 and 4) possessing higher potencies against drug-resistant tumor cells than epothilone B, the most powerful of the naturally occurring epothilones, were identified. PMID- 11048952 TI - At the maize/Agrobacterium interface: natural factors limiting host transformation. AB - BACKGROUND: Agrobacterium tumefaciens has been successfully harnessed as the only natural vector for the incorporation of foreign genes into higher plants, but its use in the grain crops is often limited. Low transformation efficiency has been partly attributed to a failure in the initial events in the transformation process, specifically in the capacity of the VirA/VirG two-component system to induce expression of the virulence genes. RESULTS: Here we show that the root exudate of Zea mays seedlings specifically inhibits virulence gene expression, determine that 2-hydroxy-4,7-dimethoxybenzoxazin-3-one (MDIBOA), which constitutes > 98% of the organic exudate of the roots of these seedlings, is the most potent and specific inhibitor of signal perception in A. tumefaciens mediated gene transfer yet discovered, and develop a model that is able to predict the MDIBOA concentration at any distance from the root surface. Finally, variants of A. tumefaciens resistant to MDIBOA-mediated inhibition of vir gene expression have been selected and partially characterized. CONCLUSIONS: These results suggest a strategy in which a plant may resist pathogen invasion by specifically blocking virulence gene activation and yet ensure that the 'resistance factor' does not accumulate to levels sufficient to impose toxicity and selection pressure on the pathogen. The data further establish that naturally occurring inhibitors directed against signal perception by the VirA/VirG two component regulatory system can play an important role in host defense. Finally, selected variants resistant to specific MDIBOA inhibition may now be used to extend the transformation efficiency of maize and possibly other cereals. PMID- 11048953 TI - The biosynthetic gene cluster for the antitumor drug bleomycin from Streptomyces verticillus ATCC15003 supporting functional interactions between nonribosomal peptide synthetases and a polyketide synthase. AB - BACKGROUND: The structural and catalytic similarities between modular nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs) inspired us to search for a hybrid NRPS-PKS system. The antitumor drug bleomycin (BLM) is a natural hybrid peptide-polyketide metabolite, the biosynthesis of which provides an excellent opportunity to investigate intermodular communication between NRPS and PKS modules. Here, we report the cloning, sequencing, and characterization of the BLM biosynthetic gene cluster from Streptomyces verticillus ATCC15003. RESULTS: A set of 30 genes clustered with the previously characterized blmAB resistance genes were defined by sequencing a 85-kb contiguous region of DNA from S. verticillus ATCC15003. The sequenced gene cluster consists of 10 NRPS genes encoding nine NRPS modules, a PKS gene encoding one PKS module, five sugar biosynthesis genes, as well as genes encoding other biosynthesis, resistance, and regulatory proteins. The substrate specificities of individual NRPS and PKS modules were predicted based on sequence analysis, and the amino acid specificities of two NRPS modules were confirmed biochemically in vitro. The involvement of the cloned genes in BLM biosynthesis was demonstrated by bioconversion of the BLM aglycones into BLMs in Streptomyces lividans expressing a part of the gene cluster. CONCLUSION: The blm gene cluster is characterized by a hybrid NRPS-PKS system, supporting the wisdom of combining individual NRPS and PKS modules for combinatorial biosynthesis. The availability of the blm gene cluster has set the stage for engineering novel BLM analogs by genetic manipulation of genes governing BLM biosynthesis and for investigating the molecular basis for intermodular communication between NRPS and PKS in the biosynthesis of hybrid peptide-polyketide metabolites. PMID- 11048954 TI - Site-directed mutagenesis of squalene-hopene cyclase: altered substrate specificity and product distribution. AB - BACKGROUND: Two regions of squalene-hopene cyclase (SHC) were examined to define roles for motifs posited to be responsible for initiation and termination of the enzyme-catalyzed polyolefinic cyclizations. Specifically, we first examined the triple mutant of the DDTAVV motif, a region deeply buried in the catalytic cavity and thought to be responsible for the initiation of squalene cyclization. Next, four mutants were prepared for Glu45, a residue close to the substrate entrance channel proposed to be involved in the termination of the cyclization of squalene. RESULTS: The DDTAVV motif in SHC was changed to DCTAEA, the corresponding conserved region of eukaryotic oxidosqualene cyclase (OSC), by the triple mutation of D377C/V380E/V381A; selected single mutants were also examined. The triple mutant showed no detectable cyclization of squalene, but effectively cyclized 2,3-oxidosqualene to give mono- and pentacyclic triterpene products. Of the Glu45 mutants, E45A and E45D showed reduced activity, E45Q showed slightly increased activity, and E45K was inactive. A normal yield of pentacyclic products was produced, but the ratio of hopene 2 to hopanol 3 was significantly changed in the less active mutants. CONCLUSIONS: Initiation and substrate selectivity may be determined by the interaction of the DDTAVV motif with the isopropylidene of squalene (for SHC) and of the DCTAEA motif with the epoxide of oxidosqualene (for OSC). This is the first report of a substrate switch determined by a central catalytic motif in a triterpenoid cyclase. At the termination of cyclization, the product ratio may be largely controlled by Glu45 at the entrance channel to the active site. PMID- 11048955 TI - Mechanism of RNase T1: concerted triester-like phosphoryl transfer via a catalytic three-centered hydrogen bond. AB - BACKGROUND: The microscopic events of ribonuclease (RNase) catalyzed phosphoryl transfer reactions are still a matter of debate in which the contenders adhere to either the classical concerted acid-base mechanism or a more sequential triester like mechanism. In the case of RNase A, small thio-effects of the nonbridging oxygens have been invoked in favor of the classical mechanism. However, the RNase T1 catalyzed transphosphorylation of phosphorothioate RNA is highly stereoselective. R(P) thio-substituted RNA is depolymerized 60000 times faster than S(P) thio-substituted RNA by this enzyme, whereas the uncatalyzed cleavage of both substrates occurs at comparable rates. We combined site-directed mutagenesis in the RNase active site and stereospecific thio-substitution of an RNA substrate to probe the intermolecular interactions of the enzyme with the nonbridging pro-S(P) oxygen that bring about this stereoselectivity of RNase T1. RESULTS: Thio-substitution of the nonbridging pro-S(P) oxygen in the substrate afflicts chemical turnover but not ground state binding whereas thio-substitution of the nonbridging pro-R(P) oxygen does not affect the kinetics of RNase T1. Site directed mutagenesis of the catalytic base Glu58 impairs the enzyme's ability to discriminate both phosphorothioate diastereomers. Glu58Ala RNase T1 cleaves R(P) and S(P) phosphorothioate RNA with similar rates. The dependence of the pro-S(P) thio-effect on the presence of the Glu58 carboxylate evidences a strong rate limiting interaction between the nonbridging pro-S(P) oxygen and the catalytic base Glu58 in the wild type enzyme. CONCLUSIONS: Based on these results, we put forward a new triester-like mechanism for the RNase T1 catalyzed reaction that involves a three-centered hydrogen bond between the 2'-OH group, the nonbridging pro-S(P) oxygen and one of the carboxylate oxygens of Glu58. This interaction allows nucleophilic attack on an activated phosphate to occur simultaneously with general base catalysis, ensuring concerted phosphoryl transfer via a triester like mechanism. PMID- 11048956 TI - Adrenergic nerves compensate for a decline in calcium buffering during ageing. AB - 1. The ubiquitous involvement of intracellular calcium ([Ca2+]i) in multiple neuronal pathways has led investigators to suggest that dysfunction of calcium homeostasis may be the primary mediator of age-related neuronal degeneration. Recently, it was shown that sympathetic neurones from superior cervical ganglion (SCG) of aged rats demonstrate decreased sarco-/endoplasmic reticulum Ca2+-ATPase (SERCA) function and that aged neurones are more dependent upon mitochondria to control K+-evoked [Ca2+]i transients. 2. Therefore, in the present study we investigated age-related changes in ATP-dependent calcium pumps of plasma membrane Ca2+-ATPase (PMCA) and SERCA in acutely dissociated SCG cells from Fischer-344 rats aged 6 and 20 months. To distinguish between PMCA and SERCA pump activity, we applied the Ca2+-ATPase blocker vanadate and measured rates of recovery of K+-evoked [Ca2+]i transients by fura-2 microfluorometry. 3. Young SCG cells showed a biphasic response to vanadate over the vanadate concentration range (0.01-100 microM); however, old SCG cells showed only a single response over the same concentration range. Additionally, old SCG cells showed a greater sensitivity to Ca2+-ATPase blockade by vanadate. 4. The contribution of mitochondrial calcium uptake to regulate [Ca2+]i was also investigated. To measure the impact of mitochondrial calcium uptake, PMCAs and SERCAs were blocked with vanadate (100 microM) and extracellular sodium was replaced with tetraethylammonium (TEA) to block Na+/Ca2+-exchange. Treated SCG cells showed a decline of 50% in rate of recovery of [Ca2+]i in both 6- and 20-month-old cells; however, this effect did not vary with age. 5. These data suggest that there is an age-related decline in function of SERCAs, with an increased reliance on PMCAs to control high K+-evoked [Ca2+]i transients. In addition, there appears to be no age-related change in the capacity of the mitochondria to restore [Ca2+]i transients to basal levels. PMID- 11048957 TI - The effect of sorbinil, an aldose reductase inhibitor, on aortic function in control and streptozotocin-induced diabetic rats. AB - 1. The present study investigates the effect of treatment of 14-day streptozotocin-diabetic rats with the aldose reductase inhibitor, sorbinil, on changes ex vivo in aortic vasoconstriction and vasodilation. 2. Maximum contractile responses and aortic sensitivity to phenylephrine were significantly enhanced in aortae from 14-day diabetic rats, in accordance with our previous data. 3. Endothelium-dependent relaxations to carbachol were, in contrast, depressed, although endothelium-independent relaxations to forskolin and sodium nitroprusside were unaltered. 4. Sorbinil treatment of diabetic animals failed to prevent any of these diabetes-induced alterations in aortic function, and indeed exacerbated some of these alterations. In addition, sorbinil treatment caused altered aortic responses in control animals, which sometimes mirrored those found in diabetic animals. 5. It can be concluded that sorbinil may have actions in addition to, and independent of, polyol pathway inhibition. Thus, sorbinil may not be an effective tool for the investigation of aldose reductase inhibition within the vascular system of the rat. PMID- 11048958 TI - Influence of alpha-adrenoceptor blockade on antigen- and propranolol-induced bronchoconstriction in guinea-pigs in vivo. AB - 1. Beta-adrenoceptor antagonists, such as propranolol, can provoke severe bronchoconstriction only in asthmatic subjects. Recently, we developed a guinea pig model of propranolol-induced bronchoconstriction (PIB) and the purpose of this study was to investigate the role of alpha-adrenergic nerve pathways in this reaction. 2. Phentolamine administered after an antigen challenge did not inhibit PIB; however, its administration before the antigen challenge significantly inhibited the antigen-induced bronchoconstriction and also bronchoconstriction induced by methacholine inhalation. 3. We conclude that the alpha-adrenergic nerve system is not involved in the development of PIB following allergic reaction in our guinea-pig model. PMID- 11048959 TI - Lack of Ca2+- and ATP-dependent priming stage in caffeine-induced exocytosis in bovine adrenal chromaffin cells: comparison with Ca2+. AB - 1. Caffeine (20-40 mM) secreted catecholamines from beta-escin-permeabilized bovine adrenal chromaffin cells in the presence or absence of 2 mM MgATP. The caffeine-induced catecholamine secretion in the presence of MgATP was to the same extent as that in the absence of MgATP. 2. Ca2+ (0.1-10 microM) induced a significantly greater secretion of catecholamines in the presence of MgATP than in the absence of MgATP. 3. ML-9 (100 microM) and ML-7 (100 microM), myosin light chain kinase inhibitors, and W-7 (100 microM) and trifluoperazine (TFP; 30 microM), calmodulin antagonists, inhibited the Ca2+-induced catecholamine secretion in the presence of MgATP but not in the absence of MgATP. They did not inhibit the caffeine-induced catecholamine secretion in the presence of MgATP. 4. The ATP-independent phase in Ca2+-dependent exocytosis is thought to be associated with the final step that ultimately leads to fusion, while the ATP dependent phase is thought to be associated with a vesicle priming reaction. Therefore, these results suggest that the ATP-requiring priming stage is lacking in the process of caffeine-induced exocytosis in bovine adrenal chromaffin cells. PMID- 11048960 TI - Renal kallikrein-kinin system, but not renal dopamine system, mediates the natriuretic response to intravenous saline infusion in healthy Chinese subjects. AB - 1. To assess the role of renal dopamine (DA), sympathetic nervous system (SNS) activity and the renal kallikrein-kinin system in sodium excretion in Chinese subjects, we studied the effects of intravenous saline infusion on the urinary excretions of sodium, free DA, free noradrenaline (NA) and kallikrein in eight healthy males aged 23-25 years. 2. After a baseline period of 1 h (hour 0), these subjects received 11 of 0.9% saline over 2 h (hours 1 and 2), followed by a 4-h recovery period (hours 3-6). From hours 0-4, subjects remained in the supine position, except to void urine. Distilled water was given orally throughout the study to ensure an adequate diuresis. 3. A 31-39% increase in sodium excretion (P < 0.05) was seen during hours 2 and 3. Urinary DA did not change throughout the study period. Urinary free NA showed no changes while the subjects remained supine, but an increase of 91-105% (P < 0.02) was seen after the subjects became ambulatory. However, there was a 103-140% increase in urinary kallikrein excretion (P < 0.05) during the saline infusion. Urinary kallikrein was still much higher (by 74%) than the basal level 1 h after the completion of the saline infusion. 4. There is no evidence from the present study that renal DA or SNS play any role in the natriuretic response to saline infusion in Chinese subjects. The brisk urinary kallikrein response, despite a relatively small salt load, suggests that the renal kallikrein-kinin system may play an important role in extracellular fluid volume and sodium homeostasis in Chinese subjects. PMID- 11048961 TI - Adrenoceptor-mediated modulation of endothelial-dependent vascular smooth muscle growth. AB - 1. Signals derived from endothelial cells (EC) and the sympathetic nervous system are known to independently modulate the growth of vascular smooth muscle (VSM). It is not known if and how these signals are integrated. The purpose of the present study was to test the hypothesis that activation of adrenoceptors by sympathetic-derived catecholamines modulates EC regulation of VSM growth. 2. The effects of adrenergic agonists on VSM growth were studied in vitro in EC/VSM cocultures. EC stimulated VSM growth in EC/VSM cocultures. Activation of beta adrenoceptors inhibited this stimulation. EC stimulation of VSM growth was 225+/ 31% in the absence and 127+/-27% in the presence of 10 microM isoprenaline. Activation of alpha-adrenoceptors had no effect on EC stimulation of VSM growth in coculture. 3. Isoprenaline did not affect the growth of VSM grown in the absence of EC, suggesting that it did not inhibit EC stimulation of VSM growth by directly inhibiting VSM growth. 4. Isoprenaline did not affect EC production of growth factors, as media conditioned by EC grown in the absence or presence of isoprenaline, stimulated VSM growth to the same extent. Isoprenaline did not alter EC, VSM or EC/VSM production of the VSM growth inhibitor transforming growth factor beta-1 (TGF-beta1). 5. These data provide evidence that catecholamines, via activation of beta-adrenoceptors, can modulate EC-dependent VSM growth, and suggest that the sympathetic nervous system and EC coordinately regulate VSM growth. PMID- 11048962 TI - Probing the selectivity of allosteric modulators of muscarinic receptors at other G-protein-coupled receptors. AB - 1. The aim of the present investigation was to analyse whether three prototype allosteric modulators of ligand binding to muscarinic receptors, i.e. alcuronium, gallamine, and the alkane-bis-ammonium compound W84 (hexane-1,6-bis[dimethyl-3' phthalimidopropylammonium bromide]), may have allosteric effects on radioligand binding characteristics at other G-protein-coupled receptors, such as cerebral A1 adenosine receptors (Gi-coupled), cardiac left ventricular alpha1-adrenoceptors (Gq), and beta-adrenoceptors (Gs). 2. The modulators were applied at concentrations known to be high with regard to the allosteric delay of the dissociation of the antagonist [3H]-N-methylscopolamine (NMS) from muscarinic M2 receptors: 30 micromol l(-1) W84, 30 micromol l(-1) alcuronium, 1000 micromol l( 1) gallamine. As radioligands, we used the adenosine A1-receptor ligand [3H] cyclopentyl-dipropylxanthine (CPX), the alpha1-adrenoceptor ligand [3H]-prazosin (PRAZ), and the beta-adrenoceptor ligand (-)-[125I]-iodocyanopindolol (ICYP). Allosteric actions on ligand dissociation and the equilibrium binding were measured in the membrane fractions of rat whole forebrain (CPX) and of rat cardiac left ventricle (PRAZ, ICYP, NMS), respectively. 3. CPX and PRAZ showed a monophasic dissociation with half-lives of 5.88+/-0.15 and 12.27+/-0.46 min, respectively. In the case of CPX, neither the binding at equilibrium nor the dissociation characteristics were influenced by the allosteric agents. With PRAZ, the binding at equilibrium remained almost unaltered in the presence of W84, whereas it was reduced to 36+/-2% of the control value with alcuronium and to 42+/-2% with gallamine. The dissociation of PRAZ was not affected by W84, whereas it was moderately accelerated by alcuronium and gallamine. In the case of ICYP, the binding at equilibrium was not affected by the allosteric modulators. The dissociation of ICYP was slow, and after 3 h, more than 50% of the radioligand was still bound, so that a reliable half-life could not be calculated. ICYP dissociation was not affected by W84. In the presence of alcuronium and gallamine, the dissociation curve of ICYP revealed an initial drop from the starting level, followed by the major phase of dissociation being parallel to the control curve. 4. In summary, the allosteric action of the applied agents is not a common feature of G-protein-coupled receptors and appears to be specific for muscarinic receptors. PMID- 11048963 TI - Cell proliferation markers in the odontogenic keratocyst: effect of inflammation. AB - The immunohistochemical expression of PCNA and Ki-67 proteins and the histochemical expression of AgNORs were studied in 20 odontogenic keratocysts in order to assess the relationship between epithelial cell proliferation and inflammation within the capsule. Immunostained cells were quantified by conventional methods, and both quantitative and morphometric analyses of AgNORs were performed by TV image analysis. Non-inflamed odontogenic keratocysts showed a typical epithelial lining and inflamed odontogenic keratocysts were lined also by hyperplastic non-keratinized stratified squamous epithelium. A statistically significant increase of PCNA+ and Ki-67+ cells and of AgNOR numbers was detected in the linings of inflamed odontogenic keratocysts compared to non-inflamed lesions. The results suggest the existence of greater proliferative activity in the epithelial cells of inflamed odontogenic keratocysts, which may be associated with the disruption of the typical structure of odontogenic keratocyst linings. PMID- 11048964 TI - Enamel proteins and extracellular matrix molecules are co-localized in the pseudocystic stromal space of adenomatoid odontogenic tumor. AB - In order to examine the functional differentiation of tumor cells of adenomatoid odontogenic tumor (AOT) as ameloblasts and to determine the participation of the extracellular matrix (ECM) in the formation of its characteristic histologic architecture, tissue samples from five cases of adenomatoid odontogenic tumor were examined by immunohistochemical staining for enamel proteins and ECM molecules. Amelogenin, enamelin, laminin, heparan sulfate proteoglycan, fibronectin, collagen type IV and type V were immunolocalized within the luminal space and along the inner rim of duct-like structures. Eosinophilic hyaline droplets within the whorled or rosette masses of tumor cells showed basically the same staining pattern as the luminal contents. High columnar tumor cells that formed duct-like structures were immunopositive for amelogenin, while the staining intensity decreased with flattening of the cells, which was a result of luminal growth. The findings suggest that the constituent cells of duct-like structures are differentiated once to ameloblasts but fail to mature further due instead to increased production of ECM molecules and due to their retention in the lumina. It is possible to regard these special structures in AOT as stromal pseudocysts. PMID- 11048965 TI - Effect of phenytoin on the production of interleukin-6 and interleukin-8 in human gingival fibroblasts. AB - The in vitro effect of phenytoin (PHT) on the production of interleukin-6 (IL-6) and interleukin-8 (IL-8) in human gingival fibroblasts, challenged with or without interleukin-1beta (IL-1beta), was studied. PHT (20 microg/ml) alone increased the mRNA level for both IL-6 and IL-8, as well as synergistically enhancing the production of IL-6 and IL-8, at both transcriptional and translational level in fibroblasts challenged with IL-1beta (30 pg/ml). The stimulatory effect of PHT on IL-1beta-induced IL-6 production was strongly reduced by the specific cyclooxygenase-2 inhibitor NS-398 (1 microM). The anti inflammatory drug, dexamethasone (1 microM), abolished the production of both IL 6 and IL-8 in gingival fibroblasts challenged with PHT in the presence or absence of IL-1beta. The ability of PHT, alone as well as in combination with IL-1, to upregulate the production of IL-6 and IL-8 in human gingival fibroblasts may contribute to enhanced recruitment and activation of inflammatory cells. This effect of PHT may thereby give a prerequisite for the establishment of an interaction between cytokines and connective tissue cells in the periodontal tissue, which is suggested to lead to gingival overgrowth. PMID- 11048966 TI - Vascular remodelling in chronic inflammatory periodontal disease. AB - Periodontitis is a chronic inflammatory disease of the highly vascularised supporting tissues of the teeth. Little is known about the vascular changes in untreated advanced periodontitis. Using confocal immunofluorescence microscopy and morphometry, we defined and quantified vascular remodelling in this lesion. In the connective tissue subjacent to the altered epithelium lining of the periodontal pocket, there was a significant increase in the numerical density of vascular profiles, primarily accounted for by vessels > or = 25 microm in diameter. In addition, vascular basement membranes were thickened and there was accumulation of non-vascular basement membrane remnants. We investigated the distribution of major angiogenic growth factors in periodontitis using immunohistochemistry. Basic fibroblast growth factor, although consistently associated with blood vessels, showed no regional variation in its distribution. In contrast, there was a marked regional variation in the intensity of immunostaining for vascular endothelial growth factor, with significantly reduced staining of the pocket epithelium. The changes in the vascularity of the periodontal connective tissues in untreated advanced periodontitis may be, in part, a consequence of altered expression of angiogenic activity by the epithelium. In turn, this may reflect the epithelial response to microbial flora in the microenvironment of the periodontal pocket. PMID- 11048967 TI - Detection of Helicobacter pylori DNA in recurrent aphthous stomatitis tissue by PCR. AB - Helicobacter pylori is recognised as being an aetiological agent of chronic active gastritis and peptic ulcer disease and has been associated with an increased risk of gastric cancer. The natural reservoir for H. pylori is unknown, although the oral cavity has been the focus of much attention in this respect. Given the histological similarities between gastric and oral ulceration, it seemed prudent to investigate a possible association between H. pylori and recurrent aphthous stomatitis (RAS). In this study, the potential involvement of H. pylori in the aetiology of RAS was investigated using the polymerase chain reaction (PCR). Biopsies from 28 RAS patients were analysed, in addition to 20 oral lichen planus (OLP) and 13 normal biopsies that were used as controls. Genomic DNA was extracted from biopsies, and confirmation of successful extraction of PCR-amplifiable DNA was achieved by carrying out PCR on each DNA sample with nested primers specific for the human beta-haemoglobin gene. PCR identification of H. pylori was carried out using a primer pair specific for the H. pylori 16S ribosomal RNA (rRNA) gene. Two rounds of PCR were carried out to amplify a 295-bp product, and the identity of amplified products was confirmed by DNA sequencing. H. pylori DNA was detected in 3 of 28 (11%) RAS samples but not in any of 20 OLP and 13 normal samples. These results do not support a definitive aetiological role for H. pylori in RAS, although the possibility that H. pylori may be involved in a small proportion of RAS cases cannot be excluded. PMID- 11048968 TI - Alendronate-related oral mucosa ulcerations. AB - Alendronate is widely used in the treatment of osteoporosis and other bone diseases. Although it is considered a well-tolerated drug, there are numerous reports of adverse effects on the mucosa in the upper aerodigestive tract, with oesophagitis as the most common complication. The strict regulations for the proper administration of the drug indicate that these side effects might well be the result of a direct, irritant mechanism on the upper aerodigestive tract. We present two clinical cases of patients who developed extensive palatal ulcers as a result of taking alendronate. We discuss possible mechanisms implicated in the production of the ulcers and some clinical factors of interest. PMID- 11048969 TI - Non-specific influx of T-cell receptor alpha/beta and gamma/delta lymphocytes in mucosal biopsies from a patient with orofacial granulomatosis. AB - Orofacial granulomatosis (OFG) represents an inflammatory disorder of the facial and oral mucosa, histologically characterized by non-caseating epithelioid cell granulomas. Since other granulomatous diseases have been shown to be characterized by a limited heterogeneity of alpha/beta and gamma/delta T cells, we investigated the T-cell diversity of both types of lymphocytes obtained from the same OFG patient. When we compared the T-cell receptor diversity of the lymphocytes accumulating at the site of the lesions with that of the peripheral blood counterpart, we did not find significant differences. Furthermore, no exclusive expansions of different T-cell clones were seen in the patient. From these data we conclude that, in this OFG patient, the majority of T cells have no specificity for a single or for a few antigens and that tissue accumulation of T lymphocytes is the result of a random influx of cells at the site of inflammation. PMID- 11048970 TI - Helicobacter pylori in oral aphthous ulcers. PMID- 11048971 TI - Standardized use of the AgNOR method. PMID- 11048972 TI - Histopathologic features of erythema nodosum--like lesions in Behcet disease: a comparison with erythema nodosum focusing on the role of vasculitis. AB - Many patients with Behcet disease (BD) develop lesions that clinically resemble those of erythema nodosum (EN) but differ from that condition with regard to their microscopic features. We examined 11 sections of EN-like lesions in BD and compared them with 9 sections of classic EN using routine histopathology and immunohistochemistry so as to form a comprehensive picture of the pathologic findings in BD and to determine the role of vasculitis in the formation of lesions. Erythema nodosum-like lesions of BD are characterized by panniculitis, usually lobular or mixed septal and lobular in pattern, with variable numbers of neutrophils, lymphocytes, and histiocytes as well as variable numbers of necrotic adipocytes. Vasculitis was noted in most EN-like lesions in BD. Scattered vessels showing lymphocytic vasculitis were evident in 6 sections, and foci of leukocytoclastic vasculitis were obvious in 4 sections, sometimes with phlebitis or arteriolitis. In specimens with classic EN, we did not observe vasculitis. Only the percentages of CD3+ lymphocytes and chloroacetate esterase-positive neutrophils in the infiltrating cells showed statistically significant differences (P < 0.05) between EN-like lesions in BD and EN through immunohistochemical and enzyme cytochemical studies. Because vasculitis in the EN like lesions in BD was extensive and not limited to areas of severe inflammation, we believe that it is primary vasculitis. We suggest that vasculitis is an important pathologic event in EN-like lesions in BD but cannot determine the extent to which other pathologic changes such as septal or lobular panniculitis, fat necrosis, neutrophilic infiltration, or microabscess formation are secondary features. PMID- 11048973 TI - Atypical histologic features in melanocytic nevi. AB - The atypical histologic features considered to be specific to dysplastic (atypical) nevi have been reported to occur in nevi that are common nevi by all other clinical and histologic features. The distribution and mutual relations among such features in nevi need to be further studied. Six histologic features (dimension > 5 mm, lentiginous proliferation, disordered nested pattern, melanocytic dyskaryosis, dermal lymphocytic infiltrate, suprabasal melanocytes) were analyzed in 253 melanocytic nevi with different clinical appearances. Atypical histologic features, found in 72% of nevi, occurred singly or formed numerous and highly variable combinations. Nevi formed a complex histologic spectrum comprising lesions showing a progressively increasing incidence of atypical features rather than two classes (common and dysplastic nevi). To divide the investigated lesions in objectively defined groups, we used a scoring system. In each nevus, a numeric value of 1 was assigned when each of the studied parameters was present and a value of 0 was assigned when each of these parameters was absent; on the basis of the final scores, nevi were divided in six different classes (classes 0-5). Diagnostic categories such as dysplastic nevi and common nevi seem to be inappropriate, as they do not reflect the real histologic complexity of such lesions. PMID- 11048974 TI - Histopathologic findings in cutaneous cytomegalovirus infection. AB - When cytomegalovirus (CMV) involves the skin, viral inclusions are typically present within mesenchymal cells, e.g., endothelial cells, fibrocytes, and sometimes within inflammatory cells, e.g., macrophages, in contrast to infection in other organs in which inclusions are usually present within ductal epithelial cells. Two cases of cutaneous CMV are presented, one showing prominent findings within eccrine ductal epithelium and the other revealing mostly endothelial cells affected by CMV. Due to the range of cytologic changes induced by CMV observed in these cases, there seem to be early, fully developed, and late cellular changes brought about by CMV analogous to how herpesvirus (varicella, zoster, simplex) induces different changes depending on its stage of infection. PMID- 11048975 TI - Foreign bodies in sarcoidosis. AB - Sarcoidosis is a multisystem disease of unknown etiology. The demonstration of polarizable foreign bodies in cutaneous granulomas is generally thought to exclude a diagnosis of sarcoidosis. Nevertheless. some investigators have reported systemic sarcoidosis with cutaneous manifestations in which polarizable particles were associated with granuloma formation in the skin. We searched the biopsy specimens of granulomatous lesions from 50 patients with cutaneous sarcoidosis using polarization microscopy to estimate the frequency of polarizable foreign bodies in cutaneous lesions of sarcoidosis. Using electron probe microanalysis, we sought to determine what elements compose these foreign bodies. Polarizable foreign bodies were found in the granulomatous skin lesions of 12 of 50 patients with cutaneous sarcoidosis. All 12 patients also had at least one other granulomatous systemic lesion, and 4 had biopsy specimens of a systemic lesion available for review. Polarizable foreign bodies were found in two cases. The elements identified were calcium, phosphorus, silicon, and aluminum. Polarizable foreign bodies were found in cutaneous sarcoidosis far more often than expected. Foreign bodies were also found in granulomatous systemic lesions. The foreign body may serve as an inciting stimulus for granuloma formation in selected cases of sarcoidosis. PMID- 11048977 TI - Cutaneous angiosarcoma arising in a gouty tophus: report of a unique case and a review of foreign material-associated angiosarcomas. AB - Rare cases of angiosarcoma have been reported to arise in the setting of retained foreign material or in association with arteriovenous fistulae. No previous case of angiosarcoma, or any other malignancy, has been reported to arise with a gouty tophus. We present a case of an 86-year-old man with a high-grade angiosarcoma that arose within a long-standing tophus. PMID- 11048976 TI - Immunoregulatory effector cells in drug-induced toxic epidermal necrolysis. AB - Toxic epidermal necrolysis (TEN) is a rare drug-induced disease for which the pathomechanism remains poorly understood. The effector cells of epidermal injury in TEN were studied by taking skin biopsies of early lesions in 23 TEN patients and by performing immunohistochemical tests using antibodies to factor XIIIa (type I dendrocytes), L1-protein (mainly Mac 387+ monocytes and macrophages), UCLHI (mainly CD45R0+ T-memory lymphocytes), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNFalpha). Computerized image analysis was used to evaluate the cell density relative to each immunolabeling. A statistical analysis of cellular counts revealed a numeric relation between the cell types in skin with TEN. Factor XIIIa+ dendrocytes were abundant and plump in the dermis, although Mac 387+ macrophages were the most numerous inflammatory cells in the epidermis. Their numbers greatly exceeded those of CD45R0+ T lymphocytes and cells showing immunoreactivity for either IL-6 or TNFalpha. In the epidermis, IL 6+ cells were significantly less numerous than TNFalpha+ cells. No quantitative difference was found between IL-6+ and CD45R0+ cell populations. Correlations were observed between either the numbers of TNFalpha+ cells or Mac 387+ macrophages and CD45R0+ lymphocytes. In the dermis, a significant correlation was also present between the numbers of Mac 387+ and factor XIIIa+ cells. These findings highlight the complex interactions between the inflammatory cells that mediate epidermal damage in skin with TEN. The high density of factor XIIIa+ dendrocytes and Mac 387+ macrophages in lesional skin assigns these cellular populations a prominent role in the pathomechanism of TEN. Despite a lower cell density, CD45RO+ T-memory lymphocytes likely participate in TNFalpha- and IL-6 regulated processes in the epidermis of TEN. TNFalpha seems to be a major cytokine involved in TEN, although a less prominent role can be ascribed to IL-6. PMID- 11048978 TI - Solitary primary cutaneous CD30+ large cell lymphoma of natural killer cell phenotype bearing the t(2;5)(p23;q35) translocation and presenting in a child. AB - Primary cutaneous CD30+ large cell lymphoma is an unusual tumor most commonly seen in adults. Most of these lymphomas are of T-cell origin and carry a good prognosis. We present the case of a 4-year-old girl with stage IEA CD30+ large cell lymphoma with a CD56+ natural killer cell phenotype and the t(2;5)(p23;q35) translocation. After excision, the patient has been free of disease for 44 months. Primary cutaneous CD30+ large cell lymphoma is uncommon in children. To our knowledge, primary cutaneous CD30+ natural killer type lymphoma has not been reported previously. The indolent behavior of this tumor indicates its similarity to other primary cutaneous CD30+ large cell lymphomas and its difference from other CD56+ lymphomas involving the skin, which often exhibit an aggressive clinical course. Cases such as this one illustrate why the use of a single, or even a few, immunohistochemical stains can be misleading in regard to lymphoma classification and prognostication. PMID- 11048979 TI - Immature myeloid precursors in chronic neutrophilic dermatosis associated with myelodysplastic syndrome. AB - Sweet syndrome (SS) associated with myeloproliferative disorders has been considered an inflammatory process mediated by neutrophils in which immunologic mechanisms are operative. The authors report the case of a 68-year-old man suffering from a myelodysplastic syndrome, who presented with a relapsing skin eruption resembling SS. Histopathologically, the skin infiltrates showed prominent neutrophilic features masking the underlying malignant process. Extensive immunophenotypic studies of skin revealed the presence of a few immature myeloid cells intermingled with an overwhelming infiltrate of neutrophils. The atypical cells in the skin had a phenotype identical to that of leukemic cells in the peripheral blood and bone marrow. Whether or not immature myeloid cell precursors constitute a specific infiltrate of leukemia cutis or are a result of recruitment of circulating leukemic cells to this area of inflammation is discussed. PMID- 11048980 TI - Extramedullary hematopoiesis in a pyogenic granuloma. AB - Extramedullary hematopoiesis is rare outside the setting of significant primary hematologic disease. We describe this phenomenon in an exuberant pyogenic granuloma in an otherwise healthy man. We postulate that this vascular lesion provided a suitable milieu for homing and proliferation of stem cells. PMID- 11048981 TI - Extramammary Paget disease with underlying hidradenoma papilliferum: guilt by association? AB - Extramammary Paget disease (EMPD) is a heterogenous entity representing either an intraepidermal adenocarcinoma in situ with apocrine differentiation or an expression of underlying malignancy of the skin or of the intestinal or genitourinary tract. The coexistence of EMPD with a benign underlying hidradenoma papilliferumn (HP) is, however, exceptional. We present the case of a 79-year-old woman with diffuse and patchy gray-white lesions involving her left vulva as well as an underlying 0.7-cm asymptomatic firm nodule. Histologically, the epidermis and dermis showed features characteristic of EMPD and HP, respectively. Malignant transformation in HP giving rise to EMPD in the overlying epithelium has been reported. In our case, however, failure to demonstrate continuity between the two lesions together with the lack of cytologic atypia, mitoses, and necrosis in the HP lends additional support to the possibility that the HP is "innocent" and that its association with EMPD is thus coincidental. A common histogenetic derivation of these two lesions from the mammary-like glands or from related germinative cells in the epidermis is suggested to explain this rare presentation. PMID- 11048982 TI - Genital cutaneous Crohn disease: two cases with unusual clinical and histopathologic features in young men. AB - Cutaneous Crohn disease, sometimes called metastatic Crohn disease or Crohn disease with cutaneous involvement, is a rare complication of Crohn disease in which granulomatous lesions involve skin separated from gastrointestinal lesions by normal tissue. We report two cases of cutaneous Crohn disease presenting in young males with erythematous, nontender swelling of the scrotum. One of the young males presented erythematous, nontender swelling of the penis as well. In one case, cutaneous Crohn disease represented the primary presentation. The original biopsy in this case showed unusual areas of degeneration of dermal connective tissue forming cystic cavities. The diagnostic biopsies in both cases showed sarcoidal granulomas with an associated superficial and deep perivascular mixed infiltrate including eosinophils. On endoscopy, both patients showed lesions of active Crohn disease in the colon. Because changes that would suggest cutaneous Crohn disease may not be present on the initial biopsy, unusual presentations and negative cultures may warrant a second biopsy. A high index of suspicion and open communication with the clinician are essential to diagnose this disease. PMID- 11048983 TI - Association of human papillomavirus type 6 with a verruciform xanthoma. AB - We report a case of verruciform xanthoma (VX) associated with human papillomavirus (HPV) in a 67-year-old male. The patient had a pale-reddish, granular and verrucous tumor on the right side of his scrotum for four years. Histopathologic examination showed typical features of VX. HPV was detected by immunohistochemistry, electron microscopy, and PCR examinations. Ultrastructural examination revealed virus-like particles of 40-50 nm in the nucleus of the upper epidermal keratinocytes. HPV type 6a DNA was detected in lesional tissue by polymerase chain reaction and sequence analysis. To the best of our knowledge, this is the first case report of VX associated with HPV. PMID- 11048984 TI - Necrobiotic xanthogranuloma presenting as a solitary tumor. AB - We report on a 60-year-old overweight white woman who presented with an asymptomatic flat, hard, yellow-brown subcutaneous plaque on her right hip. A total excision was performed. Histopathologic examination showed all the major features of a necrobiotic xanthogranuloma (NX) involving the entire dermis and subcutis, including areas of necrobiosis with cholesterol clefts, granulomatous infiltrate with some bizarre giant cells, numerous Touton cells, foamy cells, lymphoid follicles with germinal centers, foci of plasma cells, and "Touton cell panniculitis." A laboratory investigation revealed only slightly increased titers of cholesterol, strong positivity of anti-Borrelia antibodies, and diffuse skeletal osteoporosis with fractures of seven vertebrae. After 4 years of close follow-up, the osteoporosis was improved, and there were no signs of paraproteinemia, malignancy, or new skin lesions. The authors suggest that this case could represent a solitary clinical variant of NX without paraproteinemia. PMID- 11048985 TI - A thickened basement membrane is a clue to...lichen sclerosus! PMID- 11048986 TI - Pestilence narratives in classical literature: a study in creative imitation. II. Virgil, Ovid, Seneca, and Silius Italicus. AB - This article further explores patterns and traditions in classical accounts of pestilence. Recurring themes and expressions show that classical authors often copied earlier narratives, although literary analysis reveals that they sometimes embellished or distorted facts for their own purposes. PMID- 11048987 TI - Unusual neurotropism. PMID- 11048988 TI - Review of select transplant subpopulations at high risk of failure from standard immunosuppressive therapy. AB - Despite improvements in short-term graft and patient survival rates for solid organ transplants, certain subgroups of transplant recipients experience poorer clinical outcome compared to the general population. Groups including pediatrics, African-Americans, diabetics, cystic fibrosis patients, and pregnant women require special considerations when designing immunosuppressive regimens that optimize transplant outcomes. Problems specific to pediatric transplant recipients include altered pharmacokinetics of immunosuppressive drugs, such as cyclosporine (CsA) and tacrolimus (poor absorption, increased metabolism, rapid clearance), the need to restore growth post-transplantation, and a high incidence of drug-related adverse effects. African-Americans have decreased drug absorption and bioavailability, high immunologic responsiveness, and a high incidence of post-transplant diabetes mellitus. Diabetics and cystic fibrosis patients exhibit poor absorption of immunosuppressive agents, which may lead to underimmunosuppression and subsequent graft rejection. Pregnant women undergo physiologic changes that can alter the pharmacokinetics of immunosuppressives, thus requiring careful clinical management to minimize the risks of either under- or overimmunosuppression to mother and child. To achieve an optimal post transplant outcome in these high-risk patients, the problems specific to each group must be addressed, and immunosuppressive therapy individualized accordingly. Drug formulation greatly impacts upon pharmacokinetics and the resultant level of immunosuppression. Thus, a formulation with improved absorption (e.g., CsA for microemulsion), higher bioavailability, and less pharmacokinetic variability may facilitate patient management and lead to more favorable outcomes, especially in groups demonstrating low and variable bioavailability. Other strategies aimed at improving transplant outcome include the use of higher immunosuppressive doses, different combinations of immunosuppressive agents, more frequent monitoring, and management of concurrent disease states. PMID- 11048989 TI - Influence of religious and spiritual values on the willingness of Chinese Americans to donate organs for transplantation. AB - The rate of organ donation among minority groups in the United States, including Chinese-Americans, is very low. There is currently very little data in the biomedical literature that builds on qualitative research to quantify the attitudes of Chinese Americans toward organ donation. The present study quantitatively assesses the religious and cultural reasons that Chinese-Americans appear to be less willing to donate their organs than other populations. It also seeks to determine whether Confucian, Buddhist, or Daoist ideals are a significant factor in their overall reluctance to donate organs among respondents in this sample. A questionnaire distributed to Chinese American adults asked about general feelings toward organ donation and Buddhist, Confucian, Christian, Daoist, and other spiritual objections. The results suggest that Chinese Americans are indeed influenced by Confucian values, and to a lesser extent, Buddhist, Daoist, and other spiritual beliefs, that associate an intact body with respect for ancestors or nature. Another significant finding is that the subjects were most willing to donate their organs after their deaths, to close relatives, and then in descending order, distant relatives, people from their home country, and strangers. This 'negotiable' willingness has enormous implications for clinicians, who may be able to increase organ donation rates among Chinese Americans by, first, recognizing their diverse spiritual beliefs, and, second, offering a variety of possibilities for the organ procurement and allocation. PMID- 11048990 TI - Bone loss after renal transplantation: role of hyperparathyroidism, acidosis, cyclosporine and systemic disease. AB - In order to determine risk factors for bone loss after renal transplantation, dual energy X-ray absorptiometry was performed in 125 renal transplant patients. The bone mineral density (BMD) was expressed as a percentage of the normal population (BMD%) and Z-score (SD from normal). The whole body, lumbar spine and femoral neck BMD% (Z-score) values were 93.9 +/- 8.9 (-0.90 SD), 91.6 +/- 14.9 ( 0.98 SD) and 87 +/- 15.3 (-1.0 SD)%, respectively. Low BMD% was associated with low creatinine clearance ( < 40 mL/min: 91.6 +/- 7.9, > 40 mL/min: 95.6 +/- 8.0, p < 0.01), repeated graft loss (0: 94.4 +/- 9.1, > 1: 87.4 +/- 9.3, p < 0.05), long dialysis duration ( < 1 yr: 95.2 +/- 7.9, > 5: 90.1 +/- 10.6, p < 0.05), acidosis (bicarbonate < 21 mmol/L: 89.6 +/- 8.0, > 27: 96.7 +/- 7.2, p < 0.01), secondary and tertiary hyperparathyroidism ( < 50 ng/L: 95.9 +/- 7.1, > 200: 87.7 +/- 5.0, p < 0.01), raised alkaline phosphatase ( < 200 units/L: 95.7 +/- 7.2, > 300: 85.6 +/- 13.2, p < 0.001), osteocalcin ( < 50 microg/L: 95.2 +/- 6.7, > 100: 89.3 +/- 7.6, p < 0.01) and urinary deoxypyridinoline (< 5 nM/mM creatinine: femoral neck 89.6 +/- 10.7, > 10: 82.1 +/- 20.1, p < 0.05), low 25-OH-vitamin D ( < 10 microg/L: 91.3 +/- 9.8, > 20: 96.9 +/- 7.4, p < 0.001) and high cyclosporine concentration (0 ng/L: 98.3 +/- 7.0, > 150: 92.1 +/- 9.3, p < 0.05). Patients with clinical atherosclerosis (91.7 +/- 8.6 vs. 95.4 +/- 8.8, p < 0.01), hypoalbuminemia ( < 550 micromol/L: 87.6 +/- 13.2, > 550: 94.2 +/- 7.8, p < 0.01), renovascular disease (89.7 +/- 5.7 vs. 95.0 +/- 5.7, p < 0.05) and diabetic nephropathy (femoral neck 76.6 +/- 8.8 vs. 89.3 +/- 15.1, p < 0.01) had lower bone masses. High bone mass was associated with previous dialysis alphacalcidol therapy (0: 92.2 +/- 7.5, > 3 microg/wk: 97.3 +/- 6.9, p < 0.05). No relationships with transplantation duration, 1,25-OH-vitamin D, aluminium, calcium or steroid dose were found. No involutional changes in tertiary hyperparathyroidism could be discerned. CONCLUSION: The major threats to bone mass after renal transplantation appear to be ongoing hyperparathyroid bone disease, low renal function, acidosis, systemic disease and hypo-vitaminosis D. PMID- 11048991 TI - Partial venous thrombosis of the pancreatic allografts after simultaneous pancreas-kidney transplantation. AB - Despite new advances in transplantation, complete venous thrombosis (VT) of the pancreas after simultaneous pancreas kidney (SPK) transplantation usually results in graft loss. Data are limited regarding the outcome and treatment of partial VT of the pancreas allograft. From July 1994 to December 1999, 126 patients with IDDM/end-stage renal disease underwent SPK with systemic bladder drainage at the University of Miami. We retrospectively reviewed our experience regarding the outcome and treatment options of partial VT of the pancreas allografts. From July 1994 to April 1997, partial VT was not seen in the first 66 SPK patients induced with anti-CD3 rnAb and oral or intravenous (i.v.) tacrolimus (TAC) in the operating room. From May 1997 to June 1999, 14 (29%) out of 48 patients had VT. These cases were identified following the i.v. use of TAC with anti-IL-2R antibody-induction therapy (7/15) or without (7/33). Partial thrombosis of the splenic vein (PTSV) was documented in 10 patients, 2 had complete thrombosis of the splenic vein (CTSV), 1 had partial thrombosis of the superior mesenteric vein (PTSMV), and 1 patient had PTSV and PTSMV. These were identified incidentally during routine color Doppler ultrasonography (CDU). None of these SPK recipients demonstrates a change in clinical parameters. The first 8 patients were systemically heparinized, followed by oral anticoagulation, except 1 patient with CTSV. He progressed to complete thrombosis of the pancreas allograft and was treated with percutaneous thrombectomy and urokinase infusion, followed by heparinization and oral anticoagulation. One patient required exploration for bleeding. In an attempt to reduce the morbidity of heparinization, we treated the next 6 patients with PTSV with aspirin followed by serial CDU. All 14 patients had preservation of the endocrine and exocrine pancreatic functions. CDU showed resolution with recanalization of the thrombosed vein(s). From July 1999 to December 1999, 12 SPK recipients were administered TAC orally with or without induction therapy with anti-IL-2R antibody. So far, in this group, VT has not been identified. In summary, a total of 14 out of 126 patients (11%) had isolated VT with a mean follow-up of 36.4 months. Based on our experience, we suggest that extensive VT after pancreas transplantation, including splenic and superior mesenteric VT, be treated with heparin and subsequent oral anticoagulation for 3 months. For more limited, partial splenic VT, aspirin may be sufficient. Follow up CDU is critical for a successful outcome. The i.v. use of TAC appears to be a risk factor for the increased incidence of VT. Currently, using IL-2rmAb as induction, TAC is started orally on postoperative days 3 or 4 and aspirin on postoperative day 2. PMID- 11048992 TI - Detection of alloantibodies against non-HLA antigens in kidney transplantation by flow cytometry. AB - The presence of alloantibodies against human leucocyte antigen (HLA)-I and HLA-II antigens has been associated with hyperacute and accelerated graft rejections. However, occasionally these rejections occur in patients without donor-specific anti-HLA antibodies, suggesting the presence of other antigenic complexes that are shared by the graft and other cell populations. Usually, these antibodies are not routinely studied and their role in graft rejection is poorly understood. For this reason, we tested, by flow cytometry, the presence of panel-reactive alloantibodies (PRA) using different cell populations in 30 pre-transplant sera of kidney graft recipients. The patients studied had or had not hyperacute and accelerated rejection episodes (HARE) and did not have alloantibodies against HLA of their specific donors. We found that IgG and IgM alloantibodies directed against HLA-I antigens, different to the HLA-I antigens of the specific donors, as well as IgG against endothelium/monocyte antigens, IgM against platelets, and IgM against T cells are significantly associated with HARE, independently of the percentage of PRA. Our findings suggest that the detection of antibodies by flow cytometry against non-major histocompatibilty complex antigens may be useful as a pre-transplant crossmatch in living related donor kidney transplants to diminish the incidence of HARE. PMID- 11048993 TI - A prospective economic evaluation of basiliximab (Simulect) therapy following renal transplantation. AB - BACKGROUND: Immunoprophylaxis with basiliximab (Simulect), an anti-interleukin-2 receptor (anti-IL-2R; CD25) chimeric monoclonal antibody, has been demonstrated to significantly reduce the incidence of acute cellular rejection in adult renal allograft recipients (32% vs. placebo, p < 0.01). METHODS: An economic evaluation was conducted as part of a U.S. multi-center, randomized, double-blind, placebo controlled clinical trial comparing basiliximab plus dual immunosuppressive therapy (cyclosporine modified [Neoral] and corticosteroids) to dual therapy alone. Healthcare resources utilized by the 346 subjects in the 'intent-to-treat' population were prospectively collected over the 1-yr study period. Direct medical costs were determined for all hospitalizations, outpatient provider visits, procedures (excluding the initial transplant procedure), laboratory and diagnostic tests, and immunosuppressants, including basiliximab when administered. RESULTS: Total first-year medical costs were lower for the basiliximab group than for the placebo group ($28 927 vs. $32 300, difference = $3373). although this difference was not statistically significant. First-year hospital costs for treating acute rejection were also lower for the basiliximab group ($9328 vs. $10761, difference = $1433); however, this difference did not achieve statistical significance. Importantly, the efficacy analysis demonstrated a significant reduction in the incidence of acute rejection (38 vs. 55%, p < 0.01) in the basiliximab arm, and this was accomplished without increasing the overall cost of care. Fewer basiliximab-treated patients (8 vs. 15%,, p = 0.03) were hospitalized. This observation suggested less serious illness and reduced treatment costs among basiliximab-treated patients, because the overall incidence of infection was similar between the groups. The adverse event profile of patients receiving basiliximab was clinically and economically indistinguishable from that of those treated with placebo. CONCLUSION: Induction immunosuppression with basiliximab, combined with cyclosporine modified and corticosteroids, was therapeutically beneficial and contained medical costs during the initial post transplant year. PMID- 11048994 TI - Co-infection of two beta-herpesviruses (CMV and HHV-7) as an increased risk factor for 'CMV disease' in patients undergoing renal transplantation. AB - The ubiquity of human cytomegalovirus (CMV) and human herpesvirus-7 (HHV-7), as well as activation of these viruses during immunosuppression, allows the suggestion that both viruses could participate in the development of 'CMV disease' in patients after renal transplantation (RT). The aim of our research was to study the prevalence of latent CMV and HHV-7 infections in patients before RT, to determine interaction between these viruses in dual infection and possible association of their reactivation with the progression of 'CMV disease' after RT. Peripheral blood samples were collected from 49 patients before and up to 10-12 wk after RT. The methods used for diagnostics of viral infections were: serology, nested polymerase chain reaction (nPCR) analysis of peripheral blood leukocytes (PBL) and plasma, and virus isolation in cell cultures (morphological changes, nPCR analysis of cellular and cell-free samples, indirect immunofluorescence analysis). Before RT, CMV and HHV-7 DNAs were detected in PBL but not in the plasma samples, which indicates the presence of latent viral infection in patients. Latent dual (CMV + HHV-7) infection was prevalent (51.0%) in 49 patients, while CMV and HHV-7 infections alone were detected in 26.5 and 12.2% of patients, respectively. Risk of viral disease after RT, for recipients with latent dual infection before RT, was 12- and 2.2-fold higher in comparison with CMV and HHV-7 infections alone, respectively. Frequency of dual infection in 18 recipients with 'viral syndrome' or 'CMV disease' after RT was reliably higher (13/18, 81.3%) than CMV (1/18, 6.2%) (p < 0.025) and HHV-7 (2/18, 12.5%) (p < 0.025) infections alone. HHV-7 reactivation preceded CMV reactivation in 77.0% of the cases of dual infection in the recipients with viral disease and reactivation of both viruses preceded the development of viral disease. Severe 'CMV disease' developed in 2 out of 2 recipients with CMV primary infection and 'viral syndrome' in 1 recipient with CMV reinfection. The reactivation of CMV was detected in all recipients prior to onset of the disease. Correlation was shown between reactivation of latent HHV-7 infection and development of febrile syndrome in 2 out of 2 recipients with HHV-7 infection alone. Taking into account that dual infection is an increased risk factor for 'viral syndrome' and 'CMV disease' development, screening diagnostic should include testing for both viral infections in transplant donors as well as in recipients before and after RT. PMID- 11048995 TI - The role of percutaneous biopsy in detection of pancreatic transplant rejection. AB - The purpose of this study was to investigate the effectiveness and safety of percutaneous pancreatic transplant biopsy guided by ultrasound alone or with a combination of computerized tomography (CT) for pancreas localization and ultrasound for needle placement. We also compare our finding on the use of 18 gauge and 20-gauge needles for percutaneous pancreatic transplant biopsy. In 42 attempted biopsies performed on 21 patients, two different imaging modalities were used. Twenty-seven attempted biopsies were performed under the guidance of ultrasound alone, and 15 used a combination of ultrasound and CT. Of the 27 ultrasound-guided biopsies. 24 produced at least one sample adequate for histopathological analysis for an 89% biopsy success rate. Of the 15 biopsies guided by combined ultrasound and CT, 11 produced adequate samples for a 73% success rate. For all biopsies, an 83% success rate was found. In assessing the use of 18-gauge versus 20-gauge needles, 86 out of 110 tissue cores were adequate for histopathological analysis for a 78% yield. In 27 biopsy attempts using the 18-gauge needle, 75 tissue cores were obtained, for an average of 2.8 cores per biopsy. Fifty-seven pancreas samples collected using the 18-gauge needle were adequate for pathological evaluation for a 76% yield. With 15 biopsy attempts using the 20-gauge needle, 35 tissue cores were collected, for an average of 2.3 cores per biopsy. Twenty-nine pancreas specimens obtained from using the 20-gauge needle were adequate for analysis for an 83% yield. No major complications occurred. Only one incidence of minor complication was reported for a 2% complication rate. The only complication was local, mild bleeding at the biopsy site in one case. Air within the transplant pancreas as revealed by post-biopsy scans and streaky density appearing adjacent to the biopsy site occurred in a total of four cases and were not included. No complications were reported that required any invasive intervention. We conclude that percutaneous biopsy guided by ultrasound is a safe, simple, and effective method to detect pancreatic transplant rejection. Our results for biopsies compare favorably with other reported techniques in terms of effectiveness, complication rates, and ease of use. With its high success rate and low complications, ultrasound-guided percutaneous biopsy is an excellent method to sample pancreatic transplant. PMID- 11048996 TI - Glomerulonephritis is the major cause of proteinuria in renal transplant recipients: histopathologic findings of renal allografts with proteinuria. AB - The proteinuria in renal allograft recipients has been regarded as a sign of poor prognosis. The causes of post-transplant proteinuria include chronic rejection, chronic transplant glomerulopathy, glomerulonephritis (GN), acute rejection, and cyclosporine nephrotoxicity. Among them, chronic rejection is known to be most frequent. We analyzed the histopathologic findings of renal allograft biopsies in 197 Korean recipients with proteinuria. Among them, 26 patients developed proteinuria over 500 mg/d. All patients received baseline immunosuppression with cyclosporine. From 26 patients with post-transplant proteinuria, 29 biopsies were performed and their histologic diagnoses were immunoglobulin A nephropathy (IgAN) in 17, IgAN combined with chronic allograft nephropathy in 1, focal segmental glomerulosclerosis in 2, crescentic GN in 1, membranous GN in 1, diabetic nephropathy in 1, acute tubulointerstitial nephritis in 1, and chronic rejection in 3 biopsies. The remaining two biopsies showed nonspecific findings. The most common cause of post-transplant proteinuria was IgAN (62% of biopsies). The incidence of chronic rejection was relatively low and predominant cyclosporine associated changes were not observed. In conclusion, our data suggest that the main causes of post-transplant proteinuria in Korea are primary glomerulonephritides rather than chronic rejection or cyclosporine nephrotoxicity, and the kidney allograft biopsies from patients with proteinuria should be handled as native kidney. PMID- 11048997 TI - Combined pancreas and kidney transplantation improves survival in patients with end-stage diabetic nephropathy. AB - The purpose of this study was to find out whether prolonged normoglycemia, as achieved by a successful pancreas transplantation, can improve survival in patients with insulin-dependent diabetes mellitus. A retrospective analysis of actual 10-yr patient survival rates was done for all renal graft recipients who were given transplants more than 10 yr ago but within the cyclosporin era (i.e. 1981-1988). The actual 10-yr patient survival rate in non-diabetic renal graft recipients was 72%, In recipients of pancreas and kidney grafts and with prolonged function of the pancreas graft, the survival rate was 60%, whereas in patients subjected to simultaneous pancreas and kidney transplantation, but where the pancreatic grafts failed within 2 yr, the survival rate was 33%. In diabetic recipients of kidney transplants alone, the survival rate was 37%. The patient survival rate was substantially higher in non-diabetic patients and patients with functioning pancreas grafts compared with diabetic patients with kidney transplants alone or with failed pancreas grafts. We speculate that the decrease in mortality was due to the beneficial effect of long-term normoglycemia on diabetic late complications. PMID- 11048998 TI - The effect of Daclizumab in a high-risk renal transplant population. AB - INTRODUCTION: African-American (AA) renal transplant recipients have a higher incidence of acute rejection when compared to Caucasian renal transplant recipients. This higher rejection rate holds true even with the addition of several of the newer immunosuppressive agents (e.g. mycophenolate mofetil (MMF) and Rapamycin). Acute rejection rates among Hispanic (H) renal transplant recipients are higher in some settings, while lower or the same as in Caucasians in other settings. IL-2 receptor antibodies have been shown to decrease rejection rates when added to a regimen of cyclosporine (CsA), azathioprine and prednisone. Limited data are available on these agents in conjunction with triple CsA, MMF and prednisone therapy, particularly in higher risk group patients. We studied the effect of the addition of the IL-2 receptor antibody Daclizumab to a CsA, MMF, prednisone regimen in a group of African American and high-risk Hispanic renal transplant recipients. METHODS: This was a non-randomized, prospective study. A total of 49 renal transplant recipients (29 African American and 20 Hispanic) were studied and followed. A simultaneous cohort of 56 (31 African American and 25 Hispanic) renal transplant recipients receiving CsA, MMF and prednisone with no standard induction agent served as the control group. The study cohort received the same regimen with the addition of Daclizumab at 1 mg/kg for five doses over 10 wk. Multivariate analysis was performed to isolate independent factors influencing the study's results. RESULTS: A total of 56 patients in the control group and 49 patients in the Daclizumab group received an average follow-up of 17.1 +/- 6.9 and 12.7 +/- 5.1 months, respectively. Acute rejection rates were lower in the Daclizumab group as compared to the control group 26.4% versus 49.3% per patient years, respectively. A total of eight recurrent rejections in 6 patients occurred in the control group and none in the Daclizumab arm. Graft loss at this follow-up was no different between the groups. CONCLUSION: The addition of Daclizumab to a regimen of CsA, MMF and prednisone decreases acute rejection episodes in a high-risk group of African American and Hispanic renal transplant recipients. PMID- 11048999 TI - The enigma of deep-body thermosensory specificity. AB - Information provided by the analysis of peripheral cold and warm receptors may be considered a useful guide for assessing the specificity of thermal information originating in deep-body tissues. A wealth of data concerning the location of deep-body thermosensors and their neuronal correlates and modes of transduction permits the following theses to be proposed. 1. Unlike the peripheral warm and cold receptors, deep-body thermosensors are only in part represented by afferent fibers, mostly warm sensitive ones that are not characterized in detail, as the source of thermal information outside the central nervous system (CNS). The more important thermal information generated in the CNS originates mainly from warm sensitive neurons but contributions of cold-sensitive neurons are not definitely excluded. 2. Unlike the peripheral thermoreceptors, monomodality with respect to natural physical stimuli does not seem to be an essential property of deep-body thermosensors. By contrast, multimodality may underlie at least some of the multitude of interactions between thermoregulatory and other homeostatic control systems. 3. Temperature transduction seems to utilize molecular mechanisms that are also found in neurons that lack any thermosensory functions, and so the transduction mechanisms identified in warm-sensitive CNS neurons do not seem to be specific per se. 4. The observation of a multitude of temperature/response characteristics for thermosensitive CNS neurons has been helpful for categorizing these neurons, but there is no clear information that any one might be particularly relevant. 5. Originating from the peripheral cold and warm receptors two separate but interacting cold- and warm-signal pathways ascend multisynaptically to the hypothalamus as the highest level of thermoregulatory control, and to some extent go further to the sensory cortex. The signal contributed by a deep-body thermosensitive neuron, irrespective of its location, attains specificity by being fed properly into one of the two ascending thermosensory pathways. PMID- 11049000 TI - Winter North Atlantic oscillation effects on the tree rings of the Italian beech (Fagus sylvatica L.). AB - Climatic signals in beech tree-ring width series from Central Italy have been studied over different periods of time. Prewhitened tree-ring chronologies respond mainly to summer precipitation and they do not correlate in a significant manner with the winter North Atlantic oscillation (NAO) index. In this high frequency pattern the NAO signs are only found on a small number of rings characterized by being very narrow or wide. By contrast, tree-ring width chronologies in which all the frequency components are conserved were significantly related to the NAO. The significant inverse correlation between actual measurements of ring width and NAO is a consequence of the availability of water in the soil at the beginning of the growing season. In fact, in the Mediterranean area the recharging of soil moisture depends on the amount of winter precipitation, which is inversely correlated with the NAO. Strong signals of winter precipitation and NAO are found in the low-frequency components of tree ring growth. PMID- 11049001 TI - Influence of wind direction on pollen concentration in the atmosphere. AB - The daily pollen concentration in the atmosphere of Badajoz (SW Spain) was analysed over a 6-year period (1993-1998) using a volumetric aerobiological trap. The results for the main pollination period are compared with the number of hours of wind each day in the four quadrants: 1 (NE), 2 (SE), 3 (SW) and 4 (NW). The pollen source distribution allowed 16 pollen types to be analysed as a function of their distribution in the four quadrants with respect to the location of the trap. Four of them correspond to species growing in an irrigated farmland environment (Amaranthaceae-Chenopodiaceae, Plantago, Scirpus, and Typha), five to riparian and woodland species (Salix, Fraxinus, Alnus, Populus, and Eucalyptus), four to urban ornamentals (Ulmus, Arecaceae, Cupressaceae, and Casuarina), and three which include the most frequent pollen grains of widely distributed species (Poaceae, Quercus, and Olea). The results show that the distribution of the sources and the wind direction play a very major role in determining the pollen concentration in the atmosphere when these sources are located in certain quadrants, and that the widely distributed pollen sources show no relationship with wind direction. In some years the values of the correlations were not maintained, which leads one to presume that, in order to draw significant conclusions and establish clear patterns of the influence of wind direction, a continuous and more prolonged study will be required. PMID- 11049002 TI - Non-random mating in classical lekking grouse species: seasonal and diurnal trends. AB - This paper is the first to integrate both field and theoretical approaches to demonstrate that fertility benefits can be a direct benefit to females mating on the classical lek. Field data collected for male sharp-tailed grouse (Tympanuchus phasianellus), a classical lekking species, revealed potential fertility benefits for selective females. Adult males and individuals occupying centrally located territories on the lek were found to have significantly larger testes than juveniles and peripheral individuals. Further, using empirical data from previously published studies of classical lekking grouse species, time-series analysis was employed to illustrate that female mating patterns, seasonal and daily, were non-random. We are the first to show that these patterns coincide with times when male fertility is at its peak. PMID- 11049003 TI - Sun avoidance in the yellow baboons (Papio cynocephalus cynocephalus) of Ruaha National Park, Tanzania. Variations with season, behavior and weather. AB - Do free-ranging baboons avoid traveling towards the sun? Sun avoidance, in addition to resource and predator locations, may influence troop movement and non random use of the home range. This paper investigates how sun avoidance, as measured by facial exposure to sunlight, influences directional choices. It hypothesizes that baboons should avoid the sun in the hot, dry season and show indifference to it in the cool, lush season. This paper also hypothesizes that baboons employ sun-avoidance behaviors more while they forage or travel to resting sites than when they travel to foraging sites or engage in active social behaviors; lastly this paper hypothesizes that sun altitude, temperature, humidity, and cloud cover influence sun-avoidance behavior. Using focal-animal techniques on 21 males from free-ranging baboon troops, I collected locational data, accurate to within 1.6 m, over 15 months. I calculated the difference between baboon bearings and the sun's azimuth in angular degrees. Both linear and circular statistics indicate that baboons put significantly (P<0.01) more than 90 degrees between their bearing and the sun's azimuth under certain conditions. Contrary to hypotheses based on the detrimental effects of insolation, baboons in the cool, lush season avoid the sun, while baboons in the hot, dry season do not. In the lush season, the extent to which baboons avoid the sun does not depend on their other behaviors. Dry-season baboons demonstrate stronger sun avoidance while resting than when engaged in other behaviors. Finally, in the dry season, temperature drives sun avoidance; humidity drives it in the lush season. PMID- 11049004 TI - Atmospheric control of Aedes aegypti populations in Buenos Aires (Argentina) and its variability. AB - The mosquito Aedes aegypti is the main urban vector responsible for the transmission of dengue fever and dengue hemorrhagic fever. The city of Buenos Aires, Argentina, is located at the southern end of the world distribution of the species. The population abundance of Ae. aegypti is mainly regulated by environmental factors. We calculated the potential number of times that a female could lay eggs during its mean life expectancy, based on potential egg production and daily meteorological records. The model considers those variables implying physical hazard to the survival of Ae. aegypti, mosquito flying activity and oviposition. The results, obtained after calibration and validation of the model with field observations, show significant correlation (P<0.001) for different lags depending on the life stage. From these results, more favorable atmospheric conditions for Ae. aegypti reproduction (linked to the urban climatic change) can be observed. The climatic variability in the last decade resembles conditions at the end of 19th century. PMID- 11049005 TI - An investigation into the efficacy of intravenous diclofenac in post-operative dental pain. AB - OBJECTIVE: To evaluate the efficacy of single doses of intravenous diclofenac sodium (25, 50 and 75 mg) in patients with post-operative pain after third-molar surgery in a randomised, placebo-controlled study. METHODS: Two hundred and sixty nine patients (168 females) who required the removal of their impacted third molars participated in the study, which had received prior ethical approval. Surgery was completed under general anaesthesia and, during the early post operative period, patients received either a single intravenous dose of diclofenac (25, 50 or 75 mg) or matched placebo in random, double-blind order. Pain intensity was assessed on 10-cm visual analogue scales at fixed time points throughout a 4-h investigation period. Other efficacy variables obtained included time until rescue medication and overall assessment at 4, 6, 12 and 24 h after dosing. RESULTS: Throughout the 4-h investigation period, patients treated with diclofenac reported significantly less pain than those treated with placebo (P < 0.001). No differences were observed among the three doses of diclofenac (P = 0.22). Similar results were observed at 6, 12 and 24 h after dosing. Significant differences were also noted between the placebo group and all the diclofenac treatment groups with respect to time until rescue medication (P < 0.001) and the proportion of patients taking such medication. CONCLUSION: Single doses of i.v. diclofenac (25, 50 and 75 mg) provide significant pain relief after third-molar surgery. The efficacy of this preparation does not appear to be dose related. PMID- 11049006 TI - Driving under light and dark conditions: effects of alcohol and diazepam in young and older subjects. AB - OBJECTIVES: Driving at night time increases accident risk due to visual conditions, fatigue and impaired performance. In addition, the use of alcohol and benzodiazepines may enhance the risks related to night-time driving. We studied these aspects of traffic safety in a simulated driving test with young and older drivers. METHODS: In a double-blind, crossover, placebo-controlled study, nine young subjects, aged 22-24 years, performed simulated driving in both 'light' and 'dark' conditions, before and 1.5 h and 4 h after 0.8 g x kg(-1) ethanol (EOH) or 15 mg diazepam (DZ). Further, nine older subjects, aged 55-77 years, were similarly tested, but their EOH dose was 0.7 g x kg(-1) and the DZ dose was 10 mg. The tests were vigilance assessment on visual analogue scales (VAS), simulated driving under light and dark conditions for 6 min each and digit symbol substitution (DSS). RESULTS: In the young subjects, both EOH and DZ similarly impaired DSS, with DZ causing more subjective drowsiness, clumsiness, mental slowness and poor overall performance than EOH. During simulated driving, both EOH and DZ impaired simple and complex tracking (EOH > DZ) and prolonged reaction times (EOH = DZ). Impairment of performance was practically identical under light and dark conditions. In the older subjects, objective performance on DSS was poorer (-30%) than that of the young ones, and subjective impairment was marginal. During simulated driving, the baseline levels of variables in older subjects showed definite impairment (errors +100% to +500%) when compared with young subjects. Active drugs impaired several variables (EOH > DZ), but the statistical significances were fewer than in young subjects. Increase in reaction errors reached statistical significance, especially while driving in the dark. Otherwise the driving results in light and dark were not notably different. CONCLUSION: Young subjects drew good baselines but were sensitive to EOH and DZ, whilst the older subjects showed poor baselines but were less sensitive to EOH and DZ. Although the baseline driving and responses to treatments were different in young and older subjects, their driving and psychomotor impairment were unaffected by light conditions. PMID- 11049007 TI - Assay of topically administered ibuprofen using a model of post-injury hypersensitivity. A randomised, double-blind, placebo-controlled study. AB - OBJECTIVE: To develop a reliable assay for quantifying the analgesic efficacy of non-steroidal anti-inflammatory drugs (NSAIDs) using a model that is accepted as a paradigm of clinical pain. SUBJECTS: Fifteen normal subjects, all of whom were volunteers from medical school staff, took part in the study. METHODS: Capsaicin (20 microl) in solution (0.03 mg/ml) was applied to the volar surface of the forearm, and the skin was maintained at a constant temperature using a thermal stimulator. The magnitude of the surrounding area of mechanical allodynia to a brush stimulus (i.e. a clinical correlate of tenderness to touch) was assessed. Under double-blind, placebo-controlled conditions, the test was repeated using skin previously treated with ibuprofen gel or placebo. RESULTS: A close linear relationship was observed between skin temperature over a range of 30 degrees C to 40 degrees C and the area of capsaicin-induced allodynia. Ibuprofen gel significantly reduced (P < 0.004) the area of touch-evoked allodynia at a constant skin temperature of 40 degrees C. CONCLUSIONS: The thermal-facilitated adaptation of the capsaicin model described in this study represents an inexpensive and reliable assay for the effects of topical formulations of NSAID upon mechanical sensitivity. As such, it is a potential alternative to many clinical studies in which inherent confounding and bias can preclude a meaningful conclusion. PMID- 11049008 TI - Adverse effects of opioid analgesic treatment are correlated with a significant elevation in plasma epinephrine in healthy humans. AB - OBJECTIVE: The purpose of this study was to elucidate the relationship of plasma catecholamine concentrations with experienced pain intensity and analgesic effects in the setting of an experimental pain study with human volunteers. METHODS: Plasma norepinephrine and epinephrine concentrations of 12 healthy human volunteers were analysed before and during painful electrical tooth-pulp stimulation under medication using the highly potent opioid analgesic tilidine in a fixed tilidine/naloxone combination and with the non-steroidal anti inflammatory agent bromfenac. Catecholamine levels were compared with pharmacodynamic effects and reported adverse effects. RESULTS: Catecholamine levels revealed a statistically significant increase in plasma epinephrine concentrations (but not norepinephrine concentrations) 60-90 min after administration of tilidine/naloxone. This was correlated with the onset of adverse effects involving vertigo episodes in all reported cases. In contrast, there was no obvious correspondence of epinephrine or norepinephrine plasma concentrations to the experience of pain and analgesia. For comparison, under medication with the non-opioid analgesic bromfenac, only one mild adverse effects were noted, and no changes in plasma epinephrine or norepinephrine could be determined during the experimental sessions. CONCLUSIONS: It is proposed that elevated plasma epinephrine concentrations are a newly determined response to opioid-induced vertigo; this has possible clinical implications. PMID- 11049009 TI - Pharmacokinetics of (deaminohydroxy)toremifene in humans: a new, selective estrogen-receptor modulator. AB - PURPOSE: New selective estrogen-receptor modulators for the treatment and prevention of osteoporosis, cardiovascular disease and breast cancer are currently the focus of intense research. (Deaminohydroxy)toremifene (Z-2-[4-(4 chloro- 1,2-diphenyl-but-1-enyl)phenoxy]ethanol; FC-1271a) has been shown to prevent bone resorption in rats while having no or weak estrogen-like effects on the uterus, which makes it a good candidate drug for osteoporosis prevention. Our purpose here was to examine the pharmacokinetics of (deaminohydroxy)toremifene in humans included in two phase-I studies. METHODS: The first was a single-dose, dose-escalation study with 28 healthy male volunteers. Doses ranged from 10 mg to 800 mg. The second study was conducted during a 12-week period with 40 healthy, post-menopausal women, who received repeated oral doses of 25-200 mg. Standard pharmacokinetic parameters were assessed. RESULTS: In the single-dose study, time to reach peak concentration (tmax) ranged from 1.3 h to 4.0 h; peak concentration (Cmax) ranged from 15 ng/ml to 445 ng/ ml; and the estimated terminal elimination half-life (mean +/- SD; t1/2) was 24.8 +/- 7.0 h. In the repeated-dose study, tmax ranged from 1.9 h to 2.6 h at 6 weeks and from 2.5 h to 2.9 h at 12 weeks. Cmax ranged from 295 ng/ml to 1,043 ng/ml at 6 weeks and from 25 ng/ml to 1211 ng/ml at 12 weeks. The average t1/2 at all dose levels was 29.7 +/- 1.5 h (overall mean +/- SD). Strong linear correlations between the dose and Cmax and between the dose and the area under the curve were observed in both studies. CONCLUSION: Our results indicate that (deaminohydroxy)toremifene has pharmacokinetics suitable for single daily dosing. The prophylactic use of this agent in women susceptible to development of osteoporosis, cardiovascular disease and breast cancer could, therefore, be tested using a once-daily dosing schedule similar to those of other hormone-replacement therapy regimens. PMID- 11049010 TI - Differential inhibition of hepatic and duodenal sulfation of (-)-salbutamol and minoxidil by mefenamic acid. AB - OBJECTIVE: The aim of this investigation was to determine whether mefenamic acid and salicylic acid inhibit the sulfation of (-)-salbutamol and minoxidil in the human liver and duodenum, and if so, to ascertain whether the 50% inhibitory concentration (IC50) estimates are different in the two tissues. METHODS: Sulfotransferase activities were measured for 10 mM (-)-salbutamol and 5 mM minoxidil, and the concentration of 3'-phosphoadenosine-5'-phosphosulphate-[35S] was 0.4 microM. RESULTS: The IC50 estimates for (-)-salbutamol and minoxidil sulfation of mefenamic acid were 72 +/- 5.4 nM and 1.5 +/- 0.6 microM (liver), respectively, and 161 + 23 microM and 420 +/- 18 microM (duodenum), respectively. The figures for the liver were significantly lower (P < 0.0001) than those for the duodenum. The IC50 estimates for (-)-salbutamol sulfation of salicylic acid were 93 +/- 11 microM (liver) and 705 +/- 19 microM (duodenum, P < 0.0001). Salicylic acid was a poor inhibitor of minoxidil sulfation. CONCLUSION: The IC50 estimates for (-)-salbutamol sulfation of mefenamic acid and salicylic acid are lower than their unbound plasma concentrations after standard dosing, suggesting that mefenamic acid and salicylic acid should inhibit the hepatic sulfation of ( )-salbutamol in vivo. PMID- 11049011 TI - Factors affecting oral cyclosporin disposition after heart transplantation: bootstrap validation of a population pharmacokinetic model. AB - OBJECTIVE: To determine factors affecting the population pharmacokinetics of oral cyclosporin (CsA) in cardiac allograft recipients during the first 3 weeks after surgery. METHODS: Data were obtained from routine trough monitoring and from two extra samples drawn during a dosing interval on a randomly selected day. Whole blood CsA concentrations were assayed using high-performance liquid chromatography (HPLC). Approximately equal numbers of patients were prescribed Sandimmun (SAN) or Neoral (NEO) CsA formulations. Parameter values of a one compartment kinetic model with first-order absorption and elimination were sought together with the inter-patient and intra-patient variances using the NONMEM program. RESULTS: Improved fits resulted from using the following expression in the model to adjust apparent bioavailability as a function of post-operative day (POD): f= 0.2 + 10 x ABS (POD-5)/[(POD + 7) x 60]. The CsA clearance (CL/f) was found to be influenced by current body weight (WT). There was an absorption lag time of about 35 min with SAN, but zero lag time with NEO. Oral bioavailability (f) was increased by about 35% with concomitant diltiazem and about 18% with NEO. The CL/f was 10% higher during the daytime than at night. The final pharmacokinetic model was validated using 200 bootstrap samples of the original data. CONCLUSIONS: Using a validated population modelling approach, it was found that a number of factors influence the pharmacokinetics of CsA during the early postoperative period in cardiac transplant patients. These influences affecting oral bioavailability and clearance may need to be taken into account for maintaining appropriate concentrations of CsA in the bloodstream. PMID- 11049012 TI - Grapefruit juice can increase the plasma concentrations of oral methylprednisolone. AB - OBJECTIVE: To investigate whether the pharmacokinetics of orally administered methylprednisolone and plasma cortisol concentrations are affected by administration of grapefruit juice. METHODS: In a randomised, two-phase, cross over study, ten healthy subjects received either 200 ml double-strength grapefruit juice or water three times a day for 2 days. On day 3, 16 mg methylprednisolone was given orally with 200 ml grapefruit juice or water. Additionally, 200 ml grapefruit juice or water was ingested 0.5 h and 1.5 h after methylprednisolone administration. Plasma concentrations of methylprednisolone and cortisol were determined using liquid chromatography/mass spectrometry (LC/MS/MS) over a 47-h period. RESULTS: Grapefruit juice increased the total area under the plasma methylprednisolone concentration-time curve (AUC 0--infinity) by 75% (P < 0.001) and the elimination half-life (t1/2) of methylprednisolone by 35% (P < 0.001). The peak plasma concentration of methylprednisolone (Cmax) was increased by 27% (P < 0.01). Grapefruit juice delayed the time to the Cmax from 2.0 h to 3.0 h (P < 0.05). There was no significant difference in the plasma cortisol concentrations, measured after methylprednisolone administration, between the water and grapefruit juice phases. However, grapefruit juice slightly decreased the morning plasma cortisol concentrations before methylprednisolone administration (P < 0.05). CONCLUSIONS: Grapefruit juice given in high amounts moderately increases the AUC 0--infinity and t1/2 of oral methylprednisolone. The increase in t1/2 suggests that grapefruit juice can affect the systemic methylprednisolone metabolism. The clinical significance of the grapefruit juice methylprednisolone interaction is small, but in some sensitive subjects high doses of grapefruit juice might enhance the effects of oral methylprednisolone. PMID- 11049013 TI - Frequency of daily over-the-counter drug use and potential clinically significant over-the-counter-prescription drug interactions in the Finnish adult population. AB - OBJECTIVE: To explore the frequency of continuous use of over-the-counter (OTC) drugs among the Finnish adult population and the potential for harmful interactions between OTC drugs and prescribed (Rx) drugs. METHODS: Data were extracted from a 1995-1996 population-based interview survey on health care (n = 10,477, response rate 86%). The drug interaction classification system from the Swedish Drug Compendium FASS 1997 was used to identify OTC drugs likely to have clinically significant interactions with prescription drugs. Logistic regression was used to study factors related to continuous use and risks for interactions. RESULTS: Seventeen percent of the population had used OTC drugs and 15% had used OTC vitamins during the 2 days prior to the interview. Daily use of OTC drugs and of vitamins was reported by 7% and 9%, respectively. Continuous use of OTC drugs was related to older age, female gender, higher education, poor health status, long-term morbidity, psychosomatic symptoms (fatigue) and use of prescription drugs, but not to poor lifestyle. Four percent of the OTC drug users had taken drug combinations with potential for clinically significant interactions. Interactions were most common for ketoprofen (15% of ketoprofen users), ibuprofen (10%), and acetylsalicylic acid (ASA) (6%). The number of prescription drugs, long-term illness and lower education best explained the risk for interactions. CONCLUSION: Continuous use and potentially harmful Rx/ OTC drug interactions occur among OTC drug users. Further studies should be done to investigate whether potential combinations will actually lead to clinical problems. The possible interactions of non-steroidal anti-inflammatory drugs (NSAIDs) and analgesics with prescription drugs should especially be taken into account in drug information. PMID- 11049014 TI - The consumption of drugs by 75-year-old individuals living in their own homes. AB - OBJECTIVE: To examine the drug consumption and the extent of polypharmacy (defined as daily intake of three or more drugs) among 75-year-old persons living in their own homes and to point out potential problems associated with it. METHODS: Information on the intake of all drugs was collected from 492 subjects randomly selected. The subjects were interviewed at home, and their drug storage was examined. Information was also collected from the general practitioners (GPs) and from prescription databases. Database information comprised prescribed drugs used by the study population and the background population. RESULTS: Eighty-seven percent of the study population received prescribed drugs and 72% used over-the counter (OTC) drugs. Only 3% of the subjects did not take any drugs. Eighty percent of females and 60% of males used central nervous system (CNS) drugs, the most commonly used category. The subjects took on average 4.2 different prescribed drugs and 2.5 OTC drugs. Sixty percent used three or more prescribed drugs and 34% used five or more. Thirty percent used three or more OTC drugs. Seventeen percent had prescribed drugs not in use at the time of the examination in their drug storage. Twenty-five percent of the prescribed drugs were used without the GPs' knowledge. Thirty-one percent of the study population received prescribed drugs from two or more physicians. Potential drug interactions with clinical significance were found among 15.3% of the participants and were positively correlated to polypharmacy. CONCLUSION: Almost all 75-year-old persons receive drugs. The observed polypharmacy may increase drug-related risks. The discrepancies between the GPs' knowledge of their patients' medication and the actual intake may involve a potential risk. A better registration of the patients' total medication and the implementation of a common medication database for the use of all involved physicians may improve medication and reduce risks. PMID- 11049015 TI - Consequences of a change in reimbursement status on prescription patterns. PMID- 11049016 TI - An approach for the estimation of drug prescribing using the defined daily dose methodology and drug dispensation data. Theoretical considerations and practical applications. AB - OBJECTIVE: To test the hypothesis that comparison of defined daily dose (DDD) and drug user data may help to estimate drug exposure in a defined population and provide information about drug prescribing patterns. METHODS AND RESULTS: First, comparison of DDD figures with the number of apparent drug users (ADU, i.e., individuals for whom at least one prescription of the drug had been dispensed during a given time period) is demonstrated to correspond to the product of the prescribed daily dose (PDD) and the proportion of days in which the drug had been taken (days of treatment/days in a time period, D). The resulting equation (DDD/day)/ ADU in a time period = PDD x D is then applied to the analysis of different sets of drug dispensation data. Examples show that this approach may be helpful to monitor drug prescribing patterns over time. Moreover, in definite situations, it may provide reliable estimates of either PDD or D. CONCLUSIONS: Comparison of DDD and drug user data is suggested to be a cost-effective strategy to monitor drug prescribing patterns from an epidemiological perspective, which may be useful to researchers involved in drug utilisation studies as well as to health authorities for monitoring and regulatory purposes. PMID- 11049017 TI - Arsenicals in hematologic cancers. AB - Arsenic trioxide (AT) has been the object of renewed interest as a therapeutic since studies in China in the late 1980s confirmed its efficacy in the treatment of acute promyelocytic leukemia (APL). These studies have been replicated in the West, with complete remissions achieved in 80% to 90% of patients with refractory or relapsed APL. The drug has been relatively well tolerated. The dose used for treatment of APL (0.15 mg/kg/d) is approximately 50% of the maximum-tolerated dose (MTD). Common side effects have included fatigue, rash, fluid retention, and QTc-interval prolongation on electrocardiogram. A "retinoic acid syndrome," similar in its manifestations to that noted after administration of all-trans retinoic acid (RA), has been observed in APL patients. Recent studies have included dose-ranging trials to determine pharmacokinetics and the optimum schedule of administration, and studies of possible mechanisms of action. Promising future trials include combining AT with RA in the treatment of newly diagnosed APL, and broadening the range of AT therapy to other leukemias, lymphomas, multiple myeloma and some solid tumors. PMID- 11049018 TI - Gene therapy of hematologic malignancies. AB - Gene therapy offers many new and exciting treatment strategies for patients with hematologic malignancies. Through the transfer of genes into hematopoietic stem cells, one can reduce the sensitivity of myeloid cells to chemotherapy. Donor T cells can be modified to become sensitive to otherwise nontoxic prodrugs, allowing for their safer use as effectors in graft-versus-leukemia immune reactions following allogeneic transplantation. Neoplastic cells also may be modified to enhance their sensitivity to various drugs. Finally, neoplastic cells can be modified to enhance their immunogenicity using genes that encode immune stimulatory cytokines or cell surface proteins. Recent studies, for example, indicate that the stealth-like phenotype of leukemia cells can be reversed through transfer of genes such as the one encoding CD154, the ligand for CD40. A phase I clinical trial using autologous CD154-transduced leukemia cells as a cellular vaccine has provided encouraging results. Indeed, we may soon enter an era of effective gene therapy for hematologic malignancies. PMID- 11049019 TI - Manipulation of the stem cell as a target for hematologic malignancies. AB - Hematologic malignancies affect more than 80,000 patients in the United States each year. Some patients with lymphoma and leukemia are cured with conventional chemotherapy treatments. For others, autologous or allogeneic bone marrow transplantation may be the best therapeutic option. This chapter will explore novel therapies for the hematologic malignancies, using the stem cell as a target. We review work in the murine model looking at (1) the phenotype of the engrafting cells, (2) stem cell competition and host stem cells, (3) allochimerism with low-dose total body irradiation, and (4) the tolerance approach with costimulator blockade. Human data, including stem cell migration, adhesion receptor expression, and manipulations for gene therapy, are reviewed. The NOD/scid mouse model serves as a bridge between the basic bench work and human clinical trials, and we discuss applications related to umbilical cord blood and gene therapy, as well as discuss the inherent variability of this system. Finally, we address unique clinical applications in gene therapy, high dose cell transplants, minimal myeloablation, and cellular immune therapy as approaches to treatment of for patients with hematologic malignancies. PMID- 11049020 TI - New approaches to treating malignances with stem cell transplantation. AB - Stem cell transplantation has been successfully used to treat a wide variety of hematologic malignancies. New and exciting strategies being developed for use in conjunction with transplant will be useful in overcoming tumor resistance. It is now clear that a significant part of the antitumor effect of allogeneic stem cell transplantation is derived from the graft itself and is independent of the preparative regimen. Immune therapy derived from the donor's graft is uniquely suited for killing chemoresistant tumor cells and may prove to be an invaluable tool for decreasing the risk of relapse in patients with advanced disease. Among patients who have relapsed after allogeneic bone marrow transplantation (BMT), an immunologically based antitumor effect may be obtained simply by transfusing T cells obtained by leukopheresis of the original bone marrow donor. Referred to as donor leukocyte infusion (DLI), this technique has been used to obtain complete remissions in relapsed acute myeloid leukemia (AML), acute lymphocytic leukemia (ALL), multiple myeloma, non-Hodgkin's lymphoma, myelodysplastic syndrome (MDS), and chronic myeloid leukemia (CML). Another approach that uses the donor's graft to obtain a potent antitumor effect is the combination of nonmyeloablative BMT followed by immunotherapy with DLI. Numerous investigators are exploring ways of combining autologous BMT with immune therapy. Animal studies using tumor vaccines in conjunction with autologous transplantation offer a promising method for eliminating tumor. Patients undergoing autologous transplantation may have marrow that has been contaminated with tumor, which places them at a higher risk of relapse. Attempts have been made to eliminate contaminating tumor from the marrow by purging. PMID- 11049021 TI - Current uses of monoclonal antibodies in the treatment of acute leukemia. AB - Advances in the treatment of acute leukemia have been limited by both disease resistance and toxicity. Monoclonal antibodies have been used as a means of targeting therapy to malignant cells in the form of antibody-mediated cellular toxicity, radiation, or other cytotoxic agents. Anti-CD33 and anti-CD45 antibodies have been most extensively studied. Antibodies conjugated with either radioisotopes or cytotoxic moieties have been used as part of stem cell transplant regimens or as induction therapy in patients with relapsed acute myelogenous leukemia (AML), and have demonstrated antileukemic activity with acceptable toxicities. PMID- 11049022 TI - New approaches to acute lymphoblastic leukemia in adults: where do we go? AB - The optimization of conventional treatment approaches, such as chemotherapy, stem cell transplantation (SCT), and supportive care, and the exploration of new approaches will hopefully further improve the outcome of adults with acute lymphoblastic leukemia (ALL). Subgroup-adjusted treatment has already greatly improved treatment outcomes in T- and mature B-cell ALL. These approaches should be further refined, for example, in T-ALL with cyclophosphamide and cytarabine, in pro-B ALL with high-dose cytarabine (HdAC), in B-precursor ALL with high-dose methotrexate (HdM) and 6-mercaptopurine (6-MP), and in mature B-ALL with HdM and HdAC. The indications for SCT will be extended to include elderly patients undergoing allogeneic mini-transplants, and tumor eradication will be improved by better conditioning regimens such as radioimmunoconjugates and methods to induce the graft-versus-leukemia (GvL) effect, such as donor leukocyte infusions (DLI) or allogeneic mini-transplants applied after autologous transplants. Molecular therapeutic approaches, for example, those directed against the fusion protein BCR-ABL with ABL-tyrosine kinase inhibitor, are on the way to creating a new avenue for the treatment of ALL. In the future, drug resistance should be exploited as a pretherapeutic test for treatment strategies, but whether multidrug resistance modulation with available drugs will be used in ALL remains open. Evaluation of the pharmacokinetics of cytostatic drugs and the pharmacogenomics of cytostatic agents in adult ALL may contribute to the development of individualized treatment strategies with higher efficacy and lower toxicity. Minimal residual disease (MRD) evaluation is attractive in adult ALL, because it can be determined in a very high percentage of patients. It has been shown to be predictive for relapse and might be of benefit for redefinition of complete remission (CR), for determination of the efficacy of single treatment elements, and for treatment tailoring during the course of disease. New treatment approaches include also several forms of immunotherapy for B- as well as T lineage ALL; after the demonstration that such approaches are also effective in ALL, their optimal place in the treatment strategy for adult ALL can be determined. PMID- 11049023 TI - Novel therapeutic agents for the treatment of myelodysplastic syndromes. AB - Few chemotherapy agents have demonstrated activity in patients with myelodysplastic syndromes (MDS) and supportive management remains the standard of care. An increasing number of new drugs in development are being directed at specific molecular or biological targets of these diseases. Topotecan, a topoisomerase I inhibitor, has shown single-agent activity and is now being combined with other agents, including cytarabine. The aminothiol amifostine induces responses in about 30% of patients; however, its role is still being clarified. Agents that inhibit histone deacetylase and target DNA hypermethylation, thus permitting derepression of normal genes, include 5 azacytidine, decitabine, phenylbutyrate, and depsipeptide. Arsenic trioxide has demonstrated impressive activity in acute promyelocytic leukemia and preclinical data suggest the potential for activity in MDS. UCN-01 is a novel agent that inhibits protein kinase C and other protein kinases important for progression through the G1 and G2 phases of the cell cycle. Dolastatin-10 has extremely potent in vitro activity against a variety of tumor cell lines. Since its dose limiting toxicities include myelosuppression, it is being studied in acute myelogenous leukemia (AML) and MDS. Ras may play a role in MDS, and activation of this gene and its signaling pathways may require farnesylation. Several farnesyl transferase inhibitors are now available for study in patients with MDS. An increasing body of data suggests a possible role for angiogenesis in MDS, and several antiangiogenesis agents are in clinical trials, including thalidomide, SU5416, and anti-vascular endothelial growth factor (VEGF) antibodies. Development of new drugs and regimens will be facilitated by recently developed standardized response criteria. Future clinical trials should focus on rational combinations of these agents and others with the goal of curing patients with MDS. PMID- 11049024 TI - New treatment approaches for chronic myelogenous leukemia. AB - Chronic myelogenous leukemia (CML) is a myeloproliferative disease characterized by a specific translocation t(9;22)(q34;q11) that results in the transcription and translation of fusion proteins with constitutively activated tyrosine kinase activity and transduction along several signaling pathways. Identification and characterization of many of the members of this cascade of events has generated new drugs that are able to target specific segments of that chain. Most notable among these are the tyrosine kinase inhibitor compounds such as ST1571. Their activity in many CML patients who have become resistant to standard treatments such as interferon alfa or who have developed transformation into accelerated and blastic phases has recently been demonstrated in phase I clinical trials. Other agents and new drugs are being identified. This review provides a concise overview over some of these agents and their role in the treatment of CML today. PMID- 11049025 TI - Novel therapies for chronic lymphocytic leukemia in the 21st century. AB - Chronic lymphocytic leukemia (CLL) is one of the most common types of leukemia diagnosed in the Western Hemisphere. Introduction of the purine analogs has improved outcome for patients with CLL as measured by improvements in disease free survival and has made it possible to attain a complete remission in a minority of patients with this disease. The therapeutic success with the purine analogs in CLL has led to preclinical and early clinical investigation of a variety of other therapeutic agents. This, combined with advances in the understanding the genetic and immunobiology of CLL, leaves hope that CLL may become a curable disease in this century. PMID- 11049026 TI - Immunotherapeutic strategies for the treatment of plasma cell malignancies. AB - The use of immunotherapy to treat patients with plasma cell dyscrasias (PCD) such as multiple myeloma (MM) and Waldenstrom's macroglobulinemia (WM) has gained enormous interest in recent years, with considerable efforts being mounted by many investigators. These efforts have included the use of serotherapy (antibody mediated immunotherapy), vaccination strategies aimed at inducing allogeneic as well as autologous anti-MM immunity, and the use of donor lymphocyte infusions (DLIs). A number of cell surface antigens on malignant plasma cells and/or B cells in MM and/or WM patients have been proposed for use in tumor cell-targeted serotherapy, including immunoglobulin idiotype, CD19, CD20, CD38, CD54, CD138, HM1.24, and MUC1 core protein. Ongoing clinical trials are examining serotherapy targeting CD20 (in MM and WM) and CD38 (in MM), with early reports of responses to the anti-CD20 monoclonal antibody (mAb) Rituximab (Genentech, South San Francisco, CA) in patients with WM and certain patients with MM. The use of agents to induce MM- and WM-selective antigens for targeting in serotherapy has been proposed based on studies demonstrating the upregulation of CD20 by interferon-gamma (IFN-gamma), and of MUC1 core protein by dexamethasone (DEX) on malignant plasma cells. Strategies to induce allogeneic anti-MM immunity have included immunization of the marrow donor to idiotypic protein, as well as DLI. In addition, proposed immunization strategies aimed at inducing autologous immunity include vaccination with dendritic cells pulsed with MM antigens, MM cell-dendritic cell fusions, carrier-linked idiotype protein, catalytic subunit of telomerase, or DNA encoding for single-chain variable fragments (scFv) linked to a carrier protein gene. Whole tumor vaccination strategies are also being examined and include the use of MM cells transfected and/or stimulated with cytokines, costimulatory molecules, or CD40 ligand. Finally, potential obstacles to the use of immunotherapy, including the presence of resistance antigens on MM and WM tumor cells, are discussed. PMID- 11049027 TI - The biology of colorectal cancer. PMID- 11049028 TI - Screening and surveillance for colorectal cancer. PMID- 11049029 TI - A new imaging technique for colorectal cancer: positron emission tomography. PMID- 11049030 TI - Current treatment options for advanced colorectal cancer. PMID- 11049031 TI - Colorectal surgery for cancer: conventional or laparoscopic? PMID- 11049032 TI - Sphincter-preserving surgery in patients with rectal cancer. PMID- 11049033 TI - Adjuvant therapy in high-risk colon cancer. PMID- 11049034 TI - Adjuvant and neoadjuvant radiation therapy for rectal cancer. PMID- 11049035 TI - Mechanism of action of fluoropyrimidines: relevance to the new developments in colorectal cancer chemotherapy. PMID- 11049036 TI - Chronotherapy with 5-fluorouracil and other drugs in gastrointestinal malignancies. Chronotherapy Group of the European Organization for Research and Treatment of Cancer. PMID- 11049037 TI - Thymidylate synthase inhibitors in colorectal cancer. PMID- 11049038 TI - Trimetrexate as a biochemical modulator of 5-fluorouracil and leucovorin in colorectal cancer. PMID- 11049039 TI - Oral fluoropyrimidines in colorectal cancer. PMID- 11049040 TI - Oxaliplatin in colorectal cancer: an overview. PMID- 11049042 TI - Surgical resection of liver metastases of colorectal carcinoma: short and long term results. PMID- 11049041 TI - Combination chemotherapy and colorectal cancer. PMID- 11049043 TI - Cryosurgery for colorectal liver metastases. PMID- 11049044 TI - Hepatic arterial therapy for the treatment of metastatic colorectal cancer. PMID- 11049045 TI - Immunotherapy for colorectal cancer: potential application in an adjuvant setting. PMID- 11049046 TI - Colorectal cancer chemotherapy: irinotecan. PMID- 11049047 TI - Rational development of capecitabine. PMID- 11049048 TI - Future treatment options with capecitabine. PMID- 11049049 TI - Adjuvant immunotherapy in colorectal cancer. PMID- 11049050 TI - Targeting progress: the development of growth factor receptor-directed therapy. PMID- 11049051 TI - State-of-the-art chemotherapy for advanced breast cancer. AB - Several recent advances have led to accelerated progress in breast cancer therapy. The development of new drugs with novel mechanisms of action, such as the taxanes, or oral bioavailability, such as capecitabine, has expanded the horizons of available chemotherapy. The use of tumor-related proteins or genes as markers of sensitivity or resistance to systemic therapy may allow for more predictable outcomes. Finally, the emergence of biological therapies such as the HER2/neu monoclonal antibody trastuzumab (Herceptin; Genentech, Inc, So. San Francisco, CA) represents an exciting new direction that opens doors to new concepts in antitumor therapy. This report will review the most exciting possibilities for expanding the field of breast cancer management. PMID- 11049052 TI - Biological rationale for HER2/neu (c-erbB2) as a target for monoclonal antibody therapy. AB - The physical characteristics of tumor antigens that would make the most ideal targets for antibody therapeutics include cell surface expression; high, stable expression levels in tumor cells; low or absent expression in normal tissues; lack of a soluble form of the antigenic target; and lack of internalization of the antigen/antibody complex. HER2/neu is a 185-kd surface membrane protein that is overexpressed in approximately 25% of human breast cancers due to amplification of the HER2 gene. Overexpression of the gene results in ligand independent activation of HER2 kinase, causing mitogenic signal transduction and increased cell proliferation. Consequently, patients with this alteration have a worse clinical prognosis. Trastuzumab (Herceptin; Genentech, Inc, So. San Francisco, CA), a humanized anti-HER2 monoclonal antibody, has significant clinical activity against metastatic breast cancers with HER2/neu overexpression, despite the fact that the p185HER2 protein product lacks some of the ideal characteristics of tumor antigens listed above. We propose that the efficacy of trastuzumab may be explained on the basis of its effects on signal transduction, which is independent from its immune mechanism(s) of action. Furthermore, trastuzumab is synergistic with some chemotherapeutic drugs, resulting in improved therapeutic efficacy. Thus, in the case of trastuzumab, a clear distinction may be drawn between the use of monoclonal antibodies as immuneactive agents and their use to achieve a desired cellular/biochemical activity. PMID- 11049053 TI - Clinical trials of single-agent trastuzumab (Herceptin). AB - The HER2 gene (also known as neu and as c-erb-B2) encodes a 185-kd transmembrane glycoprotein receptor with intrinsic tyrosine kinase activity. HER2 is overexpressed in 25% to 30% of human breast cancers, plays a role in the pathogenesis of breast cancer, and predicts for a worse prognosis in patients with metastatic disease. Trastuzumab (Herceptin; Genentech, Inc, So. San Francisco, CA), a humanized monoclonal antibody that targets the HER2 oncogene receptor, was shown to be active in preclinical models. In initial phase I clinical trials, trastuzumab was found to be safe and to exhibit dose-dependent pharmacokinetics. Three phase II studies of single-agent trastuzumab, which was administered weekly in the outpatient setting, have now been conducted in patients with HER2-overexpressing metastatic breast cancer. In the initial phase II study, the response rate was 11% in a heavily pretreated patient population. In a pivotal follow-up study of single-agent trastuzumab, more than 200 patients who had received at least one prior chemotherapeutic regimen for metastatic disease were entered. Despite a number of unfavorable baseline characteristics, the response rate reported by an independent response evaluation committee was 15%. A more recent study in previously untreated patients has shown a 23% response rate. The median duration of response in these trials has ranged from 6.6 to 9.1 months. In these three phase II studies, trastuzumab has been shown to be safe. The most clinically significant adverse event has been cardiac dysfunction syndrome, which occurred in less than 5% of patients. Trastuzumab is not associated with the other commonly observed side effects of chemotherapy, such as alopecia, mucositis, and neutropenia. The results from these studies demonstrate that trastuzumab is active and safe in patients with metastatic HER2 overexpressing breast cancer. PMID- 11049054 TI - Current and planned clinical trials with trastuzumab (Herceptin). AB - Trastuzumab (Herceptin; Genentech, Inc, So. San Francisco, CA) is a high affinity, humanized anti-HER2 antibody that has shown benefit in the therapy of patients with metastatic HER2-overexpressing breast cancer. Results from initial clinical trials of trastuzumab as a single agent or in combination with chemotherapy established important guidelines for the therapy of HER2-positive tumors, but represent an early phase of clinical development. Preclinical studies have shown additive and synergistic effects of trastuzumab when given in combination with several chemotherapeutic agents, and a series of clinical trials exploring these new combinations is under way or will be started shortly. Substantial effort will be directed toward promising schedules and combinations, such as weekly paclitaxel with trastuzumab or combinations of platinum, taxanes, and trastuzumab. Due to the unexpected cardiac toxicity observed with the combination of doxorubicin plus trastuzumab, new studies will be designed to prospectively assess cardiac function and will incorporate anthracyclines with less cardiotoxicity, such as liposomal doxorubicin. Combination studies are not limited to cytotoxic agents, as laboratory and clinical data have demonstrated that HER2 overexpression results in resistance to hormonal therapy. Therefore, a series of studies combining hormonal treatments with trastuzumab is being considered. Finally, the integration of trastuzumab into the adjuvant and neoadjuvant settings will be studied by United States and European cooperative groups. PMID- 11049055 TI - The use of HER2 testing in the management of breast cancer. AB - Recently there has been a resurgence in interest in the use of HER2 protein overexpression or gene amplification to refine prognostic estimates of breast cancer patient outcomes and to predict which therapies might be most appropriate for individual breast cancer patients. To bring HER2 testing into clinical practice, two hurdles must be cleared. First, HER2 tests must be developed that accurately reflect HER2 status and that can be reliably performed. Second, HER2 test results must be correlated with patient outcome and response to therapy. Much progress has been made in both of these areas, but the full implementation and utility of this work have yet to be realized. PMID- 11049056 TI - The effect of fat content on the microbiology and proteolysis in cheddar cheese during ripening dairy foods. AB - We investigated the effect of incremental reduction in fat content, in the range 33 to 6% (wt/wt), on changes in the microbiology and proteolysis of Cheddar cheese, over a 225-d ripening period at 7 degrees C. A reduction of fat content resulted in significant increases in contents of moisture and protein and a decrease in the concentration of moisture in nonfat substance. Reduced fat had little effect on the age-related changes in the population of starter cells. The populations of nonstarter lactic acid bacteria decreased with fat content, and counts in the low fat cheese (6% wt/wt) were significantly lower than those in the full fat cheese (33% wt/wt) at ripening times >1 and <180 d. Proteolysis as measured by the percentage of total N soluble at pH 4.6 or in 70% ethanol decreased significantly as the fat content decreased. However, the content of pH 4.6 soluble N per 100 g of cheese was not significantly influenced by fat content. At ripening times >60 d, the content of 70% ethanol soluble N per 100 g of full fat (33% wt/wt) cheese was significantly lower than that in either the half fat (17% wt/wt) or low fat (6% wt/wt) cheeses. The concentration of AA N, as a percentage of total N, was not significantly affected by fat content. However, when expressed as a percentage of total cheese, amino acid N increased significantly with decreasing fat content. Analysis of pH 4.6 soluble N extracts by reverse phase- and gel permeation HPLC revealed that fat content affected the pattern of proteolysis, as reflected by the differences in peptide profiles. PMID- 11049057 TI - Micellar changes induced by high pressure. influence in the proteolytic activity and organoleptic properties of milk. AB - We investigated the effects of high-pressure treat-of 25 to 60 degrees C, on micelle structure, proteolytic activity, and sensory properties of milk. Pressure treatments at 25 degrees C considerably reduced micelle size, while pressurization at higher temperatures progressively increased micelle dimensions. Pressure-induced denaturation of beta-lactoglobulin (beta-LG) amounted 76% at 25 degrees C and was almost 100% in milks treated at 40 to 60 degrees C. alpha Lactalbumin (alpha-LA) was resistant to pressure at temperatures up to 40 degrees C, but its denaturation reached 56% at 60 degrees C. Plasmin resisted pressurization at room temperature; however, pressure treatments at higher temperatures increased plasmin inactivation, which reached 86.5% at 60 degrees C. Pressurization at temperatures from 40 to 60 degrees C reduced the proteolytic activity and improved the organoleptical properties of milk, compared with the same treatments at 25 degrees C, which suggested that these combined treatments could be used to produce milk of good sensory properties with an increased shelf life. These results are discussed in the light of the changes found in micellar structure. PMID- 11049058 TI - Effect of psychrotrophic bacteria and of an isolated protease from Pseudomonas fluorescens M3/6 on the plasmin system of fresh milk. AB - In the present study, we investigated the effect of Pseudomonas spp. growth on the plasmin enzymatic system in casein and whey fractions of fresh milk. Two bacterial strains, Pseudomonas spp. SRM28A and Pseudomonas fluorescens M3/6, were inoculated at a level of approximately 10(3) cfu/ml into fresh milk and incubated at 7 degrees C for 3 d. Bacterial counts were approximately 10(8) cfu/ml by d 3. Samples collected every 24 h were treated to separate the casein from the whey fraction. Casein and whey fractions were subjected to electrophoresis to visualize protein breakdown and plasmin activity and to colorimetric assays to quantify plasmin-related activities. With psychrotrophic bacterial growth, plasmin levels in casein fractions decreased significantly and in whey fractions increased then decreased significantly. Fresh milk results were similar for the two strains and were similar to earlier results with reconstituted nonfat dry milk. A transmission electron microscopy study by immunocytochemistry showed the presence of plasmin in casein micelles and its disappearance upon microbial growth in the milk. We hypothesized that extracellular microbial proteases produced by psychrotrophic microorganisms are responsible for this effect. To confirm this, an extracellular bacterial protease was isolated from Pseudomonas fluorescens M3/6 by ammonium sulfate fractionation and ion-exchange chromatography and incubated with fresh milk. Milk samples analyzed during incubation with the protease had significantly increased plasmin and plasminogen activities in the whey fraction within 5 h of incubation, while differences in activities in the casein fraction occurred at time 7.5 h for plasmin activity and 10 h of incubation for plasminogen activity. These quantitative data were supported by plasmin activity as visualized by casein-SDS-PAGE. These results suggest that growth of the Pseudomonas strains in fresh milk, and particularly their production of extracellular proteases, may be a causative factor in the release of plasmin from the casein micelle. Such plasmin release could affect the quality of cheeses and other food products that utilize dairy ingredients. PMID- 11049060 TI - Separation of lactoferrin-a and -b from bovine colostrum. AB - Bovine lactoferrin was separated into lactoferrin-a and lactoferrin-b from bovine colostrum. Lactoferrin-a was eluted at 0.38 M NaCl and lactoferrin-b was eluted at 0.43 M NaCl by carboxymethyl cation-exchange chromatography at pH 7.7, 0.05 M phosphate buffer. The molecular weights were estimated at 84,000 for lactoferrin a and 80,000 for lactoferrin-b. Lactoferrin-a contents were 258.0 mg/L and lactoferrin-b contents were 524.3 mg/L of colostrum for cow 19. From colostrum to normal milk, total lactoferrin was from 17.1 to 129.4 mg/L during the normal lactational period; however, lactoferrin did not separate clearly into lactoferrin-a and lactoferrin-b. The lactoferrin-a measured from six cows was 258.0, 114.0, 112.8, 64.0, 59.7, and 22.4 mg/ L and the lactoferrin-b 524.3, 331.8, 184.7, 170.7, 129.3, and 44.0 mg/L, respectively. The average was 105.2 mg (31.3%) for lactoferrin-a and 230.8 mg (68.7%) for lactoferrin-b. PMID- 11049059 TI - Detection of potentially allergenic material in 12 hydrolyzed milk formulas. AB - Hypoallergenic milk formulas are used as an alternative diet for infants who have allergies to cow's milk when breast-feeding is not possible. These products are based on proteins, which have been heat-treated and hydrolyzed to a different degree in order to cleave antibody-binding structures. Even extensively hydrolyzed products have occasionally been observed to elicit allergic reactions in sensitized infants, however. Therefore, the parameters of relevance to allergenic potential require more investigation. The objective of the present study was to investigate 12 different hydrolyzed milk formulas for their contents of potentially allergenic protein material, i.e. material that may induce allergenicity or elicit allergic responses in already sensitized individuals. Analytical methods applied were gel filtration, sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), native PAGE, immunoblotting, dot immunobinding, and ELISA. Care was taken to assure that all protein fractions were investigated, including supernatants and precipitates following centrifugation of the milk formulas. By gel filtration, protein material with apparent molecular masses of 7 to >30 kDa was detected. Analysis by SDS-PAGE of formula precipitates showed that proteins with a molecular mass above 20 kDa were present even in some of the extensively hydrolyzed formulas. Residual antigenic beta-lactoglobulin was found by ELISA in all products. By immunoblotting and dot immunobinding with antibodies against total whey, caseins, or Kunitz soybean trypsin inhibitor, we observed antigenic material mainly in partially hydrolyzed products. We concluded that SDS-PAGE of formula supernatants and precipitates gave the most differentiated profile of hydrolyzed formulas and that this method is well suited for screening potential allergenicity. PMID- 11049061 TI - Effect of milk fat, cocoa butter, and whey protein fat replacers on the sensory properties of lowfat and nonfat chocolate ice cream. AB - Lowfat and nonfat chocolate ice creams were made with 2.5% of milk fat, cocoa butter, or one of two whey protein-based fat replacers, Dairy Lo or Simplesse. Polydextrose was added as required so that all formulations contained the same amount of total solids. Ice cream was stored at a control temperature of-30 degrees C. Hardness, viscosity, and melting rate were measured by physical methods. Trained panelists conducted descriptive sensory analyses of the samples at 0, 6, and 12 wk. Attribute ratings were analyzed by analysis o variance with least significant difference mean separation and orthogonal contrasting. Data were also analyzed by multivariate analysis of variance with canonical variate analysis. Consumer acceptance (n = 50) did not differ among the fresh ice creams (wk 0). Ice cream containing milk fat had less intense cocoa flavor and was more resistant to textural changes over time compared with the other ice creams. Simplesse was more similar to milk fat than was Dairy Lo in its effect on brown color, cocoa flavor, cocoa character, and textural stability but was less similar in terms of thickness and mouthcoating. PMID- 11049062 TI - Rheological properties of ice cream mixes and frozen ice creams containing fat and fat replacers. AB - Ice cream mixes and frozen ice creams at milk fat levels of 12%, 8%, 6%, 6% plus a protein-based fat replacer, and 6% plus a carbohydrate-based fat replacer were evaluated for viscoelastic properties by dynamic testing with sinusoidal oscillatory tests at various frequencies. The storage modulus (G'), loss modulus (G"), and tan delta (G"/G') were calculated for all the treatments to determine changes in the viscous and elastic properties of the mixes and frozen ice creams due to fat content. In ice cream mixes, G' and G" exhibited a strong frequency dependence. The G" was higher than G' throughout the frequency range (1 to 8 Hz) examined, without any crossover, except for the 12% mix. Elastic properties of the ice cream mixes decreased as fat content decreased. Tan delta values indicated that fat replacers did not enhance the elastic properties of the ice cream mixes. In all frozen ice creams, G' and G" again showed a frequency dependence throughout the range tested (0.5 to 10 Hz). The amount of fat in ice creams and the degree of fat destabilization affected the elasticity in the frozen product. Even though the ice creams did not have significant elastic properties, when compared as a group the samples with higher fat content had higher elastic properties. The addition of protein-based and carbohydrate-based fat replacers did not enhance the elastic properties of the ice creams but did increase the viscous properties. PMID- 11049063 TI - External pudic venous reflux into the mammary vein in lactating dairy cows. AB - A representative blood sample from the mammary vein depends on the functional integrity of the valves in the external pudic vein (EPV). To determine if the EPV valves maintain blood flow into the inguinal direction during the second and subsequent lactations, we used eight lactating cows catheterized in the EPV, the lateral branch of the cranial mammary vein (MV), and the external pudic artery (EPA). The averaged daily milk yields were 25.0 +/- 1.8 kg in cows in second lactation and 31.5 +/- 2.9 kg in older cows. The relative time taken by a pulse dose of p-amino hippuric acid (PAH) injected into the EPV, to reach the EPA and the MV, was measured in a first trial. In a second trial, we assessed the extent of alteration of the mammary PAH blood concentration with blood originating from other tissues using a continuous infusion of PAH into the EPA simultaneously with blocking or not any EPV backflux. From the first experiment, the PAH injected into the EPV appeared first in the EPA and then in the MV in cows in second lactation, suggesting that blood flow was towards the inguinal region. But in a third-lactation cow, the order of appearance was reversed. In parallel, the occlusion trial demonstrated that the concentration of PAH in the MV was diluted by 14 to 39% with blood draining nonmammary tissues only in cows in third or fourth lactation. This resulting reversed flow from the EPV towards the MV would have a detrimental impact on conclusions of mammary gland metabolism studies conducted with cows in their third lactation or higher. PMID- 11049064 TI - Etiology of fatty liver in dairy cattle: effects of nutritional and hormonal status on hepatic microsomal triglyceride transfer protein. AB - We conducted three experiments to determine the effects of nutritional and hormonal status on microsomal triglyceride transfer protein (MTP) activity and mass. In experiment 1, 18 nonlactating Holstein cows, 75 d before expected calving date, in their second gestation or greater were monitored from d 75 to 55 prepartum. Cows were fed a control diet from d 75 to 62 prepartum for covariable measurements. From d 61 to 55 prepartum, six cows continued to receive the control diet, six cows were restricted to 2.3 kg of grass hay/d, and six cows were fed the control diet plus 1.8 kg of concentrate/d and 500 ml of propylene glycol given 2 times/d as an oral drench. Plasma glucose and serum insulin concentrations were highest in cows that received propylene glycol and lowest in feed restricted cows. Plasma nonesterified fatty acids (NEFA) and liver triglyceride (TG) concentrations were highest in feed restricted cows and not different between cows that received the control diet and cows that received propylene glycol. Hepatic MTP activity and mass were not affected by treatment in experiment 1. In experiment 2, bovine hepatocytes isolated from the caudate process of five preruminating Holstein bull calves were incubated with either 0, 0.5, 1.0, or 2.0 mM NEFA for 48 h. Intracellular TG increased linearly as NEFA concentration in the media increased. Concentration of NEFA in the incubation media had no effect on MTP activity or mass. There was a quadratic effect of concentration of NEFA in the incubation media on MTP mRNA. In experiment 3, bovine hepatocytes isolated from the caudate process of five preruminating Holstein bull calves were incubated with 2 mM [1-14C]oleate for 24 h to accumulate TG, followed by a 36-h period of TG depletion, during which hepatocytes were incubated with no hormone, 10 nM insulin, or 10 nM glucagon. There was no effect of insulin or glucagon on intracellular TG, MTP activity or mass. Cells incubated with no hormone had higher levels of MTP mRNA compared to cells incubated with insulin or glucagon during the depletion period. Results suggest that hepatic MTP mRNA may be affected by TG accumulation, insulin, and glucagon in vitro. However, hepatic MTP activity and mass are not affected by nutritional status of nonlactating dairy cows, TG accumulation in vitro, or insulin and glucagon in vitro. PMID- 11049065 TI - Changes in hepatic microsomal triglyceride transfer protein and triglyceride in periparturient dairy cattle. AB - We determined the relationship between microsomal triglyceride transfer protein (MTP) (activity, mass, and mRNA) and liver triglyceride concentration in 16 dairy cows (13 multiparous and three primiparous) from 27 d before expected calving (d 27) to 35 d postpartum (d 35), the time period when fatty liver is most likely to develop. In addition, dry matter intake, plasma nonesterified fatty acids (NEFA), and plasma glucose were monitored. There were no significant parity x time interactions. Dry matter intake, plasma NEFA, plasma glucose, and liver triglyceride were significantly affected by day of sampling. Dry matter intake was 10.7, 8.0, and 19.5 kg/d on d -27, 2, and 35, respectively. Plasma NEFA concentration was higher on d 2 (1113 microEq/L) compared with d -27 (201 microEq/L) and 35 (358 microEq/L). Plasma glucose concentration was 63.3, 54.3, and 57.8 mg/dl on d -27, 2, and 35, respectively. Hepatic triglyceride (TG) concentration increased from 1.8 to 11.8% liver TG (DM basis) on d -27 and 2, respectively. There was no difference between hepatic triglyceride concentration on d 2 and 35. There was a significant effect of day of sampling on hepatic MTP activity and mRNA. Hepatic MTP activity decreased from 2.08 to 1.79 nmole triolein transferred/ h per mg of microsomal protein on d -27 and 2, respectively, and increased from 1.79 to 2.17 nmole triolein transferred/h per mg of microsomal protein on d 2 and 35, respectively. Hepatic MTP mRNA increased from d -27 to 2 and remained elevated from d 2 to 35. There was no effect of day of sampling on MTP mass. There were no significant correlations between hepatic MTP activity, mass, or mRNA with either liver TG or plasma NEFA on any of the sampling days. The cause of a decrease in hepatic MTP activity and increase in mRNA on d 2 is unknown. However, the lack of correlation between MTP activity, mass, or mRNA with either liver TG or plasma NEFA on d 2 postpartum suggests that MTP probably does not play a role in the etiology of fatty liver that occurs in dairy cows at calving. PMID- 11049066 TI - Casein secretion and cytological differentiation in mammary tissue from bulls of high or low genetic merit. AB - We evaluated tissue content and secretion of milk proteins by mammary parenchymal explants prepared from tissue of mature Holstein bulls as a possible index for genetic merit. Explants from eight selection-line and eight control-line bulls were cultured for 48 or 96 h in medium with 5% fetal bovine serum with or without the addition of lactogenic hormones. Four selection-line and four control-line bulls were also treated for 7 d with estrogen and progesterone or placebo before tissue was removed on d 15 of the experiment. Overall, tissue content and secretion of casein were increased approximately twofold in selection-line bulls. Differences between genetic lines were evident only with the addition of lactogenic hormones. However, treatment of bulls with steroids was not necessary for casein secretion or detection of differences between genetic lines. Averaged for both lines, 96-h media concentrations of casein were 89-fold greater for explants cultured with added lactogenic hormones. Epithelial cells were classified as nonsecretory, droplet-containing, or degenerated. The appearance of droplet-containing cells was markedly increased in cultures supplemented with lactogenic hormones, and the cells were most often located within epithelial folds or pockets. Only occasionally did we observe areas of alveolar-like tissue. The data demonstrate that it is possible to induce the secretion of casein in mammary explants prepared from sexually mature bulls, that lactogenic hormones markedly stimulate secretion, and that differences in genetic merit may be expressed. PMID- 11049067 TI - Influence of bispecific antibodies on the in vitro bactericidal activity of bovine neutrophils against Staphylococcus aureus. AB - We conducted the following study to determine if bispecific antibodies enhance the bactericidal activity of bovine neutrophils. Bispecific antibodies were synthesized by chemically crosslinking bovine neutrophil monoclonal antibodies to Staphylococcus aureus 305 capsule polysaccharide monoclonal antibodies. The efficiency of chemically coupling monoclonal antibody monomers was approximately 50% for each bispecific antibody produced. Monoclonal antibodies against neutrophils enhanced the respiratory burst activity of neutrophils by 2.3- to 2.5 fold. To determine the influence of bispecific antibodies on neutrophil function, S. aureus 305 was preincubated with various concentrations of bispecific antibodies and neutrophils were then added to the opsonized bacteria at different bacteria to neutrophil ratios. The bactericidal activity of neutrophils was expressed as a percentage reduction in colony-forming units in test cultures compared with the number of colony-forming units in control test cultures that did not contain bispecific antibodies or neutrophils. The addition of bispecific antibodies to test cultures increased the bactericidal activity of neutrophils. A reduction in colony-forming units as a function of increasing the S. aureus 305 to neutrophils ratio was observed in both the absence and presence of bispecific antibodies. However, a greater reduction was observed in the presence of bispecific antibodies. Increasing concentrations of bispecific antibodies enhanced the bactericidal activity of neutrophils at a constant S. aureus 305 to neutrophil ratio of 1:500. The results indicate that bispecific antibodies that recognize both S. aureus 305 capsular polysaccharide and neutrophil antigens potentiate the bactericidal activity of neutrophils. PMID- 11049068 TI - A comparison of two commercially available Escherichia coli J5 vaccines against E. coli intramammary challenge. AB - The efficacy of two commercially available Escherichia coli J5 bacterins was investigated. Jersey cows were randomly assigned to one of three treatment groups: 1) unvaccinated controls, 2) vaccinated with J.VAC (Merial Limited, Athens, GA), and 3) vaccinated with J5 bacterin. All cows were vaccinated at drying off and at 2 wk before anticipated calving. Cows that were vaccinated with the J5 bacterin also received a third immunization at calving. One quarter of each cow was challenged with approximately 64 cfu of E. coli at 14 to 30 d postcalving. Immunization by either vaccine did not influence the severity of coliform mastitis; however, the mean number of colony-forming units of E. coli recovered from challenged quarters was significantly lower for immunized cows than for control cows at 144 h postchallenge. Serum and mammary secretion immunoglobulin (Ig)G, IgG1, and IgG2 titers against E. coli J5 whole-cell antigens were enhanced in vaccinated cows. Serum and mammary secretion IgM were not different among treatment groups. Somatic cell counts in milk from challenged quarters, rectal temperatures, and the clinical status of cows following intramammary challenge were not different among treatment groups. PMID- 11049069 TI - Short communication: growth hormone response to somatostatin-28 and growth hormone-releasing factor in dairy heifers. AB - In a attempt to investigate the role fo somatostatin-28 (SRIF-28) in modulating growth hormone (GH)-releasing factor (GRF)-induced GH release in dairy cattle, we assigned Holstein heifer calves (n = 12) to receive 0, 10, and 20 microg of SRIF 28/100 kg of BW (0, 10, 20) in conjunction with administration of 3 microg of SRIF-28/100 kg ofBW. Administration of 10 or 20 microg of SRIF-28 reduced GRF induced GH release, because areas under the response curves during 30 min after GRF administration were less when cows received those dosages compared with receiving 0. Some evidence of a dose response was observed because serum GH concentrations increased from 15 to 20 min after GRF administration when animals received 10 microg but continued to decrease when animals received 20 microg of SRIF-28. We conclude that, in contrast to studies with SRIF-14, pituitary GH release is very sensitive to inhibitory effects of SRIF-28 when used in as little as a fivefold molar excess. Additional experimentation is necessary to determine whether equimolar concentrations of SRIF-28 are effective in altering GRF-induced GH release. Further, these results give rise to speculation that SRIF may be involved in physiological mechanisms mediating well-documented genetic influences on GH concentrations in dairy cattle. PMID- 11049070 TI - Pasteurization of discard mycoplasma mastitic milk used to feed calves: thermal effects on various mycoplasma. AB - Discard milk from sick or antibiotic-treated cows is often used as an economical alternative to milk replacer at dairy farms. This practice poses a health risk to calves if the discard milk is from cows with mycoplasma mastitis. Mycoplasma bovis, Mycoplasma californicum, and Mycoplasma canadense are among the agents known to cause contagious mastitis in cattle and occasionally pneumonia, otitis media, or arthritis in calves. This report describes a recent outbreak of calf polyarthritis and respiratory disease on a midwest dairy farm. The farm fed discard mycoplasma mastitic milk to its calves. On-the-farm pasteurization of the discard milk to 65 degrees C for 1 h before feeding prevented additional illness in the calves. Discard milk samples were collected before and after heating and tested for mycoplasma by culture. Only samples collected before pasteurization yielded live cultures. Common mastitic mycoplasma agents were also tested for sensitivity to heat. It was determined that 65 degrees C killed M. bovis and M. californicum after 2 min of exposure, while M. canadense remained viable for up to 10 min. Exposure to 70 degrees C inactivated M. bovis and M. californicum after 1 min, but M. canadense samples were positive for up to 3 min. Thus, M. canadense appears to be more heat resistant than M. bovis and M. californicum. Heat treatment that results in the destruction of M. canadense should be used for the pasteurization of discard mycoplasma mastitic milk. PMID- 11049071 TI - In vitro methods for measuring the dry matter digestibility of ruminant feedstuffs: comparison of methods and inoculum source. AB - We conducted this study to evaluate the new in vitro system, DAISY", to determine dry matter (DM) digestibility in ruminant feedstuffs. Results from the DAISY" were compared to those obtained by the traditional Tilly and Terry method. The traditional method buffer was used for both methods. We also compared two sources of rumen inoculum from sheep and dairy cows. Seventeen different feeds were tested, grouped into roughage, concentrate, and CP supplements. The experiment was replicated on two different occasions for all feeds and the two sources of inoculum. The source of inoculum and the time at which it was collected had no effect on the in vitro DM digestibility of the feedstuffs in either of the methods. The DAISY" DM digestibility value compared well with the traditional method values for the roughage group; however, for some feedstuffs in the concentrate and CP supplement groups, the DAISY" values were significantly higher than the traditional method values. Regression analysis of the feeds that resulted in similar values with the two methods revealed that the DAISY" method can be used to predict in vitro digestibility with relatively small variation. PMID- 11049072 TI - Influence of particle size on the effectiveness of the fiber in barley silage. AB - We used eight multiparous Holstein cows in a 4 x 4 Latin square design to evaluate the effects and possible interactions between silage particle size and concentrate level on chewing activities and productivity of cows fed barley-based total mixed rations (TMR). Diets were designed with two forage-to-concentrate ratios (low forage, 45:55, high forage 55:45), combined with two theoretical chop lengths of barley silage (short = 4.68 mm and long = 18.75 mm). Diets were formulated to provide similar and above-minimum neutral detergent fiber recommended for cows in early lactation. Increasing silage particle size of the forage did not affect dry matter intake. The 3.5% fat-correct milk and fat yields trended higher for increased particle size. Percent milk protein was higher for short particle size. Increasing the concentrate levels in the diets increased proportions of milk protein and lactose, but not milk fat. Cows fed short silage spent 90 min less per day chewing and ruminating than did those on long silage. Total chewing activity per kilogram of forage intake was higher for cows on long silage compared with those on short silage diets. Although a reduction in silage particle size did not depress milk fat, rumination and chewing activity were significantly reduced. These results suggest that particle size of the silage may have dominant control over chewing activity despite adequate neutral detergent fiber intakes. PMID- 11049073 TI - Factors affecting pasture intake and total dry matter intake in grazing dairy cows. AB - We investigated the most relevant variables for estimating pasture intake and total dry matter (DM) intake in grazing dairy cows using 27 previously published studies. Variables compared were pasture allowance, days in milk, amount of forage, amount of concentrate and total supplementation, pasture allowance and supplementation interaction, fat-corrected milk, body weight (BW), metabolic BW, daily change in BW, percentage of legumes in pasture, neutral detergent fiber (NDF) contents of pasture, and NDF in pasture selected. The variables were selected using stepwise regression analysis for total DM intake and pasture DM intake. Variables selected in the total DM intake regression equation (R2 = 0.95) were pasture allowance, total supplementation, interaction of pasture allowance and supplementation, fat-corrected milk, BW, daily change in BW, percentage of legumes and pasture NDF content. Pasture DM intake regression equation (R2 = 0.90) was similar to total DM intake equation, but supplementation coefficient was negative, showing substitution effect in supplementing grazing cows. The intake of NDF as a percentage of BW was higher than 1.3% when considering NDF content of the pasture allowance. Low pasture allowance groups had values higher than 1.3%. PMID- 11049074 TI - Peripartum responses of dairy cows to partial substitution of corn silage with corn grain in diets fed during the late dry period. AB - We randomly assigned 189 cows in a commercial dairy farm to dietary treatments with supplemental corn grain (SC) or without supplemental corn grain (NC) approximately 3 wk before expected parturition. Diets formulated were similar except that dry ground corn replaced 21% of the corn silage in one diet. Cows fed SC had reduced plasma beta-hydroxybutyrate and tended to have increased plasma insulin concentrations prepartum compared with cows fed NC. Treatment did not affect nonesterified fatty acid concentrations prepartum, any blood variables postpartum, or incidences of health disorders. Effects of treatment on production responses were highly dependent on parity as indicated by parity x treatment x time interactions for milk and protein yields. Primiparous cows fed SC had lower milk protein yield, higher somatic cell count and days open compared with cows fed NC. The SC diet resulted in lower milk yields in early lactation and increased somatic cell count and days open for cows in second parity. However, cows in third parity or greater fed the SC diet yielded more milk and protein in early lactation, and had decreased somatic cell counts and days open. Increasing the corn grain concentration of the diet fed prepartum was advantageous to third and greater parity cows in this experiment, but showed no benefits during lactation for cows in first or second parities. PMID- 11049075 TI - Determination of when during the lactation cycle to start feeding a cellulase and xylanase enzyme mixture to dairy cows. AB - We used 48 multiparous Holstein cows to compare the response of dairy cows to a direct-fed mixture of cellulase and xylanase enzymes (1.25 L of enzyme concentrate/tonne of forage dry matter) applied to the forage portion (60% corn silage and 40% alfalfa hay) of a total mixed diet starting either in the close-up dry period, at calving, or at peak milk production. Cows were blocked by calving date and, within blocks, randomly assigned to one of four treatment diets. Treatments were: 1) an untreated control diet, 2) enzyme addition to the forage from wk 6 to 18 postpartum, 3) enzyme addition to the forage from calving to wk 18 postpartum, and 4) enzyme addition to the forage from wk 4 prepartum to wk 18 postpartum. Total mixed diets were 65% forage and 35% concentrate prepartum, and 50:50 forage:concentrate postpartum. The production of milk, solids-corrected milk, fat-corrected milk, and energy-corrected milk was higher for cows fed enzyme-treated diets than for cows fed control diet. Production was similar for cows in all enzyme-treated groups, although numerically highest for cows that started receiving enzyme-treated forages right after parturition and numerically lowest when started prepartum. Concentrations of fat, protein, and lactose in milk were similar for all treatments; yields of protein and fat were higher for cows fed enzyme-treated forages. Dry matter intake and body condition scores, both prepartum and postpartum, were similar for all diets. Eating rates, as determined in two 24-h studies, were similar for control and enzyme-treated diets. The feeding of enzyme-treated forages increased milk production. While the effect of when the feeding of enzyme-treated forages started was not statistically significant, we recommend starting soon after parturition because of the greatest total milk production when starting at that time. PMID- 11049076 TI - Partitioning of amino acids flowing to the abomasum into feed, bacterial, protozoal, and endogenous fractions. AB - We partitioned the flow of amino acids (AA) to the abomasum among rumen undegradable protein (RUP) and bacterial, protozoal, and endogenous fractions using four Holstein cows in midlactation that were equipped with ruminal and abomasal cannulas. A 2 x 2 factorial design with four diets, combinations of high or low ruminally degradable organic matter, and rumen degradable protein, was employed. Crude protein (CP) and AA contents of ruminal bacteria and protozoa and abomasal digesta were determined. Equations for the source compositions and in vivo flows of CP and 16 AA were then solved simultaneously with a linear program to estimate the contribution of RUP, bacterial, protozoal, and endogenous CP to AA flows. The flows of RUP and bacterial AA were not affected by diet. Low dietary RDP increased the flow of protozoal AA to the abomasum, but the ruminally degradable organic matter content of the diet did not affect protozoal AA flow. Across diets, RUP, bacterial, protozoal, and endogenous fractions provided 55, 33, 11, and <1% of the CP, and 62, 26, 12, and <1% of the AA that reached the abomasum. The linear program was a useful tool for partitioning AA that flows to the abomasum. The technique may also allow dietary effects on ruminal microbes and the AA profile of protein flowing to the duodenum to be better understood and perhaps manipulated. PMID- 11049078 TI - Application of a mixed normal mixture model for the estimation of Mastitis related parameters. AB - The current methodology for estimating genetic parameters for SCC (SCS) does not account for the difference in SCS between healthy cows and cows with an intramammary infection (IMI). We propose a two-component finite mixed normal mixture model to estimate IMI prevalence, separate SCS subpopulation means, individual posterior probabilities of IMI, and SCS variance components. The theory is presented and the expectation-conditional maximization algorithm is utilized to compute maximum likelihood estimates. The methodology is illustrated on two simulated data sets based on the current knowledge of SCS parameters. Maximum likelihood estimates of IMI prevalence and SCS subpopulation means were close to simulated values, except for the estimate of IMI prevalence when both subpopulations were almost confounded. Individual posterior probabilities of IMI were always higher among infected than among healthy cows. Error and additive variance components obtained under the mixture model were closer to simulated values than restricted maximum likelihood estimates obtained assuming a homogeneous SCS distribution, especially when subpopulations were completely separated and when mixing proportion was highest. Convergence was linear and rapid when priors were chosen with caution. The advantages of the methodology are demonstrated, and its feasibility for large data sets is discussed. PMID- 11049077 TI - Effect of dietary thiamin supplementation on milk production by dairy cows. AB - We conducted three experiments to determine the effects of dietary thiamin supplementation on milk production by dairy cows. In trial 1, 28 Holstein cows were blocked by parity and assigned randomly to either placebo or thiamin top dress for the 8-wk experiment to provide a supplemental thiamin intake of 0 or 150 mg/d per cow. Within each of these groups, cows were further assigned randomly to two total mixed rations (TMR) for 4 wk, with the TMR treatments then reversed for a second 4-wk experimental period. Milk yield was 2.7 kg/d higher for thiamin-supplemented cows. Yields of milk fat and protein were increased 0.13 and 0.10 kg/d, respectively, by dietary thiamin supplementation. In trial 2, 20 multiparous Holstein cows were used in a crossover design with 4-wk periods. Placebo or thiamin premixes were added to TMR to provide an approximate daily supplemental thiamin intake of 0 or 300 mg/cow. Milk and protein yields tended to be 0.7 and 0.04 kg/d higher, respectively, for thiamin-supplemented cows. In trial 3, 16 multiparous Holstein cows were used in a replicated 4 x 4 Latin square with 21-d periods. Placebo or thiamin premixes were added to TMR to provide an approximate daily supplemental thiamin intake of 0 or 300 mg/cow. Dry matter intake tended to be 0.8 kg/d lower for thiamin-supplemented cows. Milk fat percentage tended to be 0.18 percentage units lower and fat yield was 0.08 kg/d lower for thiamin-supplemented cows. Thiamin supplementation tended to increase milk and component production when dietary concentrations of neutral and acid detergent fiber were lower and nonfiber carbohydrate was higher than recommended. PMID- 11049079 TI - Heritability of clinical mastitis incidence and relationships with sire transmitting abilities for somatic cell score, udder type traits, productive life, and protein yield. AB - The objective of this study was to determine the relationships among daughter clinical mastitis during first and second lactations and sire transmitting abilities for somatic cell score, udder type traits, productive life, and protein yield. Data on clinical mastitis during first lactation were available for 1795 daughters (in six Pennsylvania herds, one Minnesota herd, and one Nebraska herd) of 283 Holstein sires. Data on clinical mastitis during second lactation were available for 1055 of these daughters. A total of 479 cows had 864 clinical episodes during first lactation, and 230 cows had 384 clinical episodes during second lactation. Clinical mastitis incidence and the total number of clinical episodes during each lactation were regressed on herd-season of calving (a classification variable), age at first calving, lactation length, and sire transmitting abilities taken one at a time. Linear effects, nonlinear effects, and odds ratios were estimated for sire transmitting abilities. Separate analyses were conducted on dependent variables that considered clinical mastitis from: all organisms, coagulase-negative staphylococci, coliform species, streptococci other than Streptococcus agalactiae, and the most common environmental organisms (coliform species and streptococci other than Streptococcus agalactiae). Heritability of clinical mastitis ranged from 0.01 to 0.42. Daughters of sires that transmit the lowest somatic cell score had the lowest incidence of clinical mastitis and the fewest clinical episodes during first and second lactations. Daughters of sires that transmit longer productive life, shallower udders, deeper udder cleft, and strongly attached fore udders had either fewer clinical episodes or lower clinical mastitis incidence during first and second lactations. The incidence of clinical mastitis and the number of clinical episodes per lactation may be reduced by selection for lower somatic cell score, longer productive life, shallower udders, deeper udder cleft, or strongly attached fore udders. PMID- 11049080 TI - A cost analysis of encapsulated spray-dried milk fat. AB - Anhydrous butter oil or cream was encapsulated in all-purpose flour, modified cornstarch, or sucrose and then spray-dried. We estimated the processing cost for a plant designed to produce 57,000 kg/d (125,000 lbs/ d) of encapsulated milk fat powder. Powder with a 50% milk fat content could be produced for about $0.23/kg plus the cost of the butter oil or cream, the encapsulant selected, and the other ingredients. Spray-drying of milk fat improved ease of handling and reduced storage costs. PMID- 11049081 TI - Evaluation of reproductive performance in lactating dairy cows with prostaglandin F2alpha, gonadotropin-releasing hormone, and timed artificial insemination. AB - Nine hundred and twenty Holstein cows from 16 commercial dairy herds to evaluate three systematic breeding protocols: 14-d PGF2alpha, timed artificial insemination (AI), and GnRH-PGF2alpha, relative to AI following estrus detection without hormone intervention. The timed AI protocol involved GnRH, followed by PGF2alpha 7 d later and GnRH again 2 d after PGF2alpha, with AI 6 to 18 h after the second GnRH. The GnRH-PGF2alpha protocol consisted of GnRH followed by PGF2alpha 7 d later. Eight herds relied on visual observation to detect estrus, and eight herds utilized the HeatWatch estrus detection system. The average interval to first postpartum AI was shortest for the timed AI protocol (77.1 d) followed by the 14-d PGF2alpha protocol (81.6 d). There was no difference in days to first AI between the control (86.1 d) and GnRH-PGF2alpha (89.5 d) protocols. Percent pregnant per first AI did not differ among control (45.6%), 14-d PGF2alpha (43.7%), or GnRH-PGF2alpha (44.0%) protocols, but all protocols had a higher percent pregnant per first AI than the timed AI protocol (30.1%). Response to the GnRH-PGF2alpha protocol was limited because 44.0% of the cows submitted to the protocol were not detected in estrus < or = 10 d post-PGF2, administration and had an interval to first AI of 103.8 d. Cumulative percent pregnant by 120 d postpartum did not differ between control cows (53.1%) and hormonally treated cows (50.6%). Visual observation herds had a shorter interval to first postpartum AI (82.8 d) than the HeatWatch herds (84.8 d), with a higher overall rate of estrus detection across all protocols (75.3 and 67.6%, respectively). PMID- 11049082 TI - Identification of corynebacterium bovis and other coryneforms isolated from bovine mammary glands. AB - Bovine mastitis remains the most economically important disease in dairy cows. Corynebacterium bovis, a lipid-requiring Corynebacterium spp., is frequently isolated from the milk of infected mammary glands of dairy cows and is associated with reduced milk production. A total of 212 coryneform bacteria isolated from the milk of dairy cows were obtained from mastitis reference laboratories in the United States and Canada. All isolates had been presumptively identified as Corynebacterium bovis based on colony morphology and growth in the presence of butterfat. Preliminary identification of the isolates was based on Gram stain, oxidase, catalase, and growth on unsupplemented trypticase soy agar (TSA), TSA supplemented with 5% sheep blood, and TSA supplemented with 1% Tween 80. Of the 212 isolates tested, 183 were identified as Corynebacterium spp. based on preliminary characteristics. Of the strains misidentified, one was identified as a yeast, two as Bacillus spp., 11 as Enterobacteriaceae, 18 as staphylococci, one as a Streptococcus spp., and one as an Enterococcus spp. Eighty-seven coryneforms were selected for identification to the species level by direct sequencing of the 16S rRNA gene, the Biolog system and the API Coryne system. Fifty strains were identified as C. bovis by 16S rRNA gene similarity studies: the Biolog and API Coryne systems correctly identified 54.0 and 88.0% of these strains, respectively. The other coryneforms were identified as other Corynebacterium spp., Rhodococcus spp., or Microbacterium spp. These data indicate that the coryneform bacteria isolated from bovine mammary glands are a heterogeneous group of organisms. Routine identification of C. bovis should include Gram-stain, cell morphology, catalase production, nitrate reduction, stimulated growth on 1% Tween 80 supplemented media, and beta-galactosidase production as the minimum requirements. PMID- 11049083 TI - Biosecurity on dairy operations: hazards and risks. AB - The objective here was to present a model for considering biosecurity related to infectious diseases on US dairy operations using a risk assessment framework. With the example of an important dairy cattle pathogen (Mycobacterium paratuberculosis), I followed risk assessment steps to characterize risks related to the use of certain management practices and possible risk reduction within an infectious disease biosecurity program. Biosecurity practices focus on the prevention of introduction of these pathogens to the dairy, and estimates of the risks associated with introduction of different sources of cattle are presented. In addition, biosecurity practices also limit the transmission of these pathogens within an infected dairy operation, especially those focused on sick cow management, calving area management, and manure management. Recent information from the National Animal Health Monitoring System (NAHMS) Dairy 96 Study indicates that many of these practices have not been adopted on US dairy operations, indicating both risk of disease and opportunity for animal health improvement. PMID- 11049084 TI - Photosensitization of pancreatic tumour cells by delta-aminolaevulinic acid esters. AB - A series of straight chain, branched and cyclo-delta-aminolaevulinic acid (ALA) esters have been synthesized and their photosensitizing properties analysed using an in vitro system of rat pancreatoma cells. Structurally favourable ALA esters not only induced the formation of more of the endogenous photosensitizer, protoporphyrin IX (PpIX), but they did so at a faster rate than ALA itself. This action was reflected in a substantial increase in photocytotoxicity of some 270 times, using the more potent ALA esters. An important structural feature was identified in two of the ALA esters which greatly limited PpIX production, i.e. a branch point located next to the site of ester cleavage. Experiments on the transport of ALA and of ALA esters across the cell membrane showed that ALA, but not ALA esters, gain access to the cell via the di- and tripeptide transporter, PEPTI. Finally, these results show that the esterification of ALA can greatly increase its cellular uptake, so generating more intracellular PpIX, improved tumour cell photosensitization and enhanced photocytotoxicity. PMID- 11049085 TI - Synthesis and antiproliferative activity of unsaturated quinoline derivatives. AB - In our previous work Knoevenagel condensation of quinoline 2-, 3- and 4 carbaldehyde with malononitrile derivatives was used to produce a series of heteroarylidene malononitrile derivatives. Some of these heteroaromatic tyrphostins were potent inhibitors of the epidermal growth factor (EGF) receptor kinase. This work has now been extended by using 6-, 7-, and 8 quinolinecarbaldehyde to prepare 23 new quinoline-tyrphostins 1-23. Most of these compounds were moderately active against the MCF7 breast cancer cell line. The order of potency was 7- > 6 > 8-substituted quinoline, which indicates that increased activity of the 7-substituted quinolines is associated with electron deficiency at the 7-position in the quinoline ring. The most active compound, 12, formed from 7-quinolinecarbaldehyde and ethyl cyanoacetate, had an IC50 value of 2.3 microM. Compounds 1-23 showed similar IC50 values against the MCF7 and MCF7/ADR cell lines (the latter shows fourfold increased protein tyrosine kinase activity) except for the compounds 1 and 15 formed from 6-quinolinecarbaldehyde and malononitrile and 7-quinolinecarbaldehyde and cyanoacetamide, which showed a significant (11- and 42-fold, respectively) increase in potency against the MCF7/ADR cell line. Furthermore, no association was found between growth inhibition and inhibition of the EGFR protein tyrosine kinase (PTK), using a cell free assay. In addition, new compounds were prepared from 2- and 4 quinolinecarbaldehyde with extended conjugation in the side chains (24-27) or with methoxypolyethoxyethyl esters in the side chain to increase water solubility (28 and 29). These compounds showed substantial cytotoxicity, with IC50 values in the range 1-25 microM, but similar values were observed against both cell lines. No association was found between inhibition of PTK and growth inhibition, again indicating that their mode of action may not be specific for the EGF receptor. PMID- 11049086 TI - Adriamycin-induced inhibition of mitochondrial-encoded polypeptides as a model system for the identification of hotspots for DNA-damaging agents. AB - It has recently been shown that the anti-cancer drug Adriamycin forms drug-DNA adducts which function as 'virtual' interstrand cross-links in cells, and these cross-links are specific for GpC sequences. The objective of this work was to determine whether all GpC sites are equally susceptible to the formation of Adriamycin-DNA adducts in the mitochondrial genome or whether any 'hotspots' exist whereby lesions are formed preferentially at particular GpC-containing sequences. The mitochondrial genome was used as a model system as it provides a series of contiguous genes, all of which lack introns and in which transcription is driven from a single promoter. With the absence of nucleotide excision repair, this provides an excellent system with which to observe Adriamycin-induced DNA damage since such lesions are reflected as an inhibition of mitochondrial protein synthesis. HeLa cells were treated with Adriamycin and the extent to which synthesis of individual mitochondrial-encoded proteins was inhibited was quantitated. Mitochondrial protein synthesis was found to be inhibited in a discontinuous manner, corresponding to regions rich in 5'-GpC sequences. These results therefore indicate that Adriamycin-DNA adducts do not form randomly with GpC sites throughout the mitochondrial genome, but instead appear to form preferentially at regions of high GpC content. This selective inhibition of mitochondrial-encoded proteins demonstrates the potential of this method for the in situ detection of localized regions of binding by DNA-acting drugs. PMID- 11049087 TI - DNA intercalation, topoisomerase II inhibition and cytotoxic activity of the plant alkaloid neocryptolepine. AB - Cryptolepine and neocryptolepine are two indoloquinoline alkaloids isolated from the roots of the African plant Cryptolepis sanguinolenta. Both drugs have revealed antibacterial and antiparasitic activities and are strongly cytotoxic to tumour cells. We have recently shown that cryptolepine can intercalate into DNA and stimulates DNA cleavage by human topoisomerase II. In this study, we have investigated the mechanism of action and cytotoxicity of neocryptolepine, which differs from the parent isomer only by the orientation of the indole unit with respect to the quinoline moiety. The biochemical and physicochemical results presented here indicate that neocryptolepine also intercalates into DNA, preferentially at GC-rich sequences, but exhibits a reduced affinity for DNA compared with cryptolepine. The two alkaloids interfere with the catalytic activity of human topoisomerase II but the poisoning activity is slightly more pronounced with cryptolepine than with its isomer. The data provide a molecular basis to account for the reduced cytotoxicity of neocryptolepine compared with the parent drug. PMID- 11049088 TI - Antineoplastic agents 429. Syntheses of the combretastatin A-1 and combretastatin B-1 prodrugs. AB - The original synthesis of combretastatin A-1 (1) was modified to allow an efficient scale-up procedure for obtaining this anti-neoplastic stilbene. Subsequent conversion to a useful prodrug was accomplished by diphosphorylation (to 10), with in situ formation of dibenzylchlorophosphite, followed by cleavage of the benzyl ester protecting groups with trimethyliodosilane. The phosphoric acid intermediate was treated with sodium methoxide to complete a practical route to the sodium phosphate prodrug (4). Selective hydrogenation of phosphate 10 and treatment of the product with sodium methoxide led to combretastatin B-1 prodrug (5). The phosphoric acid precursor of prodrug 4 was employed in a parallel series of reactions to produce a selection of metal and ammonium cation prodrug candidates. Each of the phosphate salts was evaluated from the perspective of relative solubility behavior and cancer cell growth inhibition. The sodium phosphate prodrug of combretastatin A-1 (4) was selected for detailed antineoplastic studies. PMID- 11049089 TI - Antimetastatic and antitumor effects of 2,4-diamino-6-(pyridine-4-yl)-1,3,5 triazine (4PyDAT) on the high lung metastatic colon 26 tumor in mice. AB - The therapeutic potential of a diaminotriazine, 2,4-diamino-6-(pyridine-4-yl) 1,3,5-triazine (4PyDAT), was investigated in a metastatic model using the mouse colon 26 carcinoma variant (Co26Lu), which preferentially metastasizes to the lung of mouse. The compound had a moderate antimetastatic activity as well as antitumor activity, without toxicity to the host, when administered orally. In the cytotoxicity test in vitro, 4PyDAT showed very weak direct cytotoxicity against the Co26Lu cell line, Co26Lu(F55) (IC50 < or = 1000 microM). Less microcapiral formation on tumors were observed for the treated group with a hemorrhage than the control group under microscopy. 4PyDAT significantly inhibited the production of urokinase-type plasminogen activator (u-PA) in Co26Lu(F55) cells. These results suggest that the antimetastatic and antitumor activities of 4PyDAT are due in part to inhibition of angiogenesis, rather than direct antiproliferative action on the tumor cells. 4PyDAT may become a lead compound to develop antitumor triazine derivatives based on antiangiogenic action. PMID- 11049090 TI - Design, synthesis and in vitro cytotoxicity studies of novel pyrrolo[2,1 c][1,4]benzodiazepine (PBD)--polymade conjugates and 2,2'-PBD dimers. AB - A series of novel pyrrolo[2,1-c][l,4]benzodiazepine (PBD)-polyamide conjugates (1 and 2) and 2,2'-PBD dimers (3, 4 and 5) were synthesized and evaluated for cytotoxicity in >60 human tumor cell lines. In general PBD-polyamide conjugates (1 and 2) exhibit higher cytotoxic potency compared with 2,2'-PBD dimers (3, 4 and 5). Compound 2 exhibits a wide spectrum of anticancer activities against 17 cell lines in six cancer panels with LC50 values of <9 microM, and is especially effective against colon cancer, melanoma, renal cancer and breast cancer. Compound 1 selectively affects cell growth against renal cancer A 498 cell line and compound 4 affects cell growth against breast cancer MDA-MB-231/ATCC cell line with an LC50 value 0.06 microM. Increases in the chain length of the linker in 2,2'-PBD dimers significantly increase the cytotoxic potency and increases in the number of pyrrole groups in the PBD-polyamide conjugates similarly increase the cytotoxic potency. PMID- 11049091 TI - VP7 and VP4 genotypes among rotavirus strains recovered from children with gastroenteritis over a 3-year period in Valencia, Spain. AB - Between September 1996 and May 1999, the incidence and distribution of the main human rotavirus G genotypes (VP7 associated: G1-G4) and P genotypes (VP4 associated: P[8], P[4], P[6] and P[9]) among children with rotavirus gastroenteritis were determined using reverse transcription and polymerase chain reaction (RT-PCR)-based genotyping methods. From a total of 145 rotavirus strains examined, we identified the G type in 131 (90.3%) and the P type in 127 (87.5%) of the samples. An overall predominance of genotypes P[8] G1 (42.7%) and P[8] G4 (32.4%) was found during the period of study, with much lower incidence of genotypes P[4] G2 (5.5%) and P[8] G3 (2%). P[6] and P[9] types were not detected, neither were unusual combinations of P and G types. A significant genotypic shift was observed: whereas P[8] G4 was the most prevalent genotype during the first year of the study (60%), the genotype P[8] G1 gradually increased to account for 62.3% of the strains analysed in the following winter season. Mixed G types revealing dual infections G1/G4 and G3/G4 were found at low frequency (2%). PMID- 11049092 TI - The changing epidemiological pattern of hepatitis A in an urban population of India: emergence of a trend similar to the European countries. AB - The present study was undertaken to determine the seroprevalence of the antibody against hepatitis A virus (IgG anti-HAV) in an urban population sample from Delhi (India) and to assess any change in the epidemiological pattern of HAV infection in this part of the world. A total of 500 healthy subjects were enrolled and divided into groups on the basis of age, sex and per capita income and evaluated for the presence of IgG anti HAV antibodies using a commercially available kit. The mean age of all the subjects was 32.6 +/- 13.2 yr. and the male:female ratio was 1.5:1. The overall prevalence of IgG anti-HAV in all subjects was 71.2% (356/500). The prevalence in subjects >35 years (92.1% [186/202]) was significantly higher than that in subjects <35 years (92.1% [186/202]) was significantly higher than in subjects <35 years (57% [170/298]). No statistically significant difference was observed between male and female subjects (71.4% [217/304] vs. 70.9% [139/196]) or between subjects belonging to middle and low socioeconomic groups (68.9% [135/196] vs. 72.7% [221/304]). These findings when compared with the results that were obtained in 1982, showed a decreasing prevalence of IgG anti-HAV, most significantly in younger age groups (16-35 years). Thus, we may conclude that the seroepidemiology of hepatitis A virus infection in urban population of India seems to be changing with seroprevalence in the younger population approaching a figure similar to that of the more developed European countries. PMID- 11049093 TI - Gender differences of symptom reporting and medical health care utilization in the German population. AB - AIM: Gender differences in morbidity have been widely confirmed in representative health surveys in North America and Europe. Significantly more women than men suffer from somatic complaints. It is less clear whether differences in symptom reporting provide an impact on health care utilization and to which degree psychosocial factors exhibit confounding influence. METHODS: We analyzed data from a representative health examination survey in Germany with 7466 participants in the age range of 25 to 69 years. RESULTS: The analysis confirmed an overall excess in female symptom reporting, both in the total sample (n = 7460; p < or = 0.001) and in the healthy subsample (n = 906, p < or = 0.01). Also, female utilization of medical services was higher (p < or = 0.0001). A simultaneous age related increase in the prevalence of symptom reporting in both groups peaked in the age group of 55-59 years followed by a subsequent slight decrease in higher age groups whereas utilization steadily increased over the adult life span in both sexes. As expected, more medical utilization was associated with higher symptom reporting levels. Nevertheless, females constantly exhibited more medical utilization than males in all symptom reporting groups. Age and marital status had no univariate influence on symptom reporting whereas low social class status (p = 0.001), poor perceived/self assessed health (p < 0.0001), and high levels of chronic distress (p < 0.0001) were associated with more symptom reporting. In multivariate analysis, the female gender lost its significance on heightened symptom reporting. Poor perceived/self assessed health had the most pronounced impact on symptom count (F-value 59.1; p < 0.001). CONCLUSIONS: The present study confirms a female excess of symptom reporting and utilization of medical services. Nevertheless, symptom reporting and utilization are not closely related. The gender gap in symptom reporting may be largely explained by low social class status, high levels of chronic distress and poor perceived/self assessed health. PMID- 11049094 TI - Potential susceptible individuals to poliomyelitis in Austria. AB - This report presents the results of a vaccine coverage survey that was performed in the Tyrol (Austria) in 1997. The major finding is that pockets of children susceptible to poliomyelitis currently exist in the area, because of delayed immunisation. The cause for the delay is the practice of interrupting oral poliomyelitis vaccine (OPV) administration during summer months. PMID- 11049096 TI - Comparing the biological and psychosocial risks of pregnancy between groups of adolescents and adults. AB - Pregnancy in adolescence constitutes the main public health problem for this age group, in some countries. The health problems derived from adolescent pregnancy, birth and neonatal attention depend more on factors of social and environmental risk than on physiological and biological risk factors in adolescence. A descriptive study has been conducted of adolescent and adult mothers in Cadiz, Spain, who attended Family Planning Clinic during 1994. By means of multistage, stratified random sampling, 590 women were selected: 305 adolescent mothers between 15 and 19 years of age, and 285 adult mothers of 20 years and over. Various questionnaires were applied to the women, covering: sociodemographic characteristics; pregnancy and birth; family and social support (Duke-Inc and Apgar family); evolution of the health of the baby; and maternal knowledge of child care. Sociodemographically, significant differences (p > 0.01) were obtained in: marital status (more unmarried among adolescents); living in parents' home; fewer working; abandonment of education. No differences were observed in respect of the number of check-ups received during pregnancy. Adult mothers consumed significantly more toxic substances (tobacco, alcohol and drugs) during pregnancy. No differences were appreciated in respect of the birth, or health of the neonate. In adolescents, there was significantly more breastfeeding (p > 0.01) compared with adult mothers, 61.6% (56-67%) and 34% (28.5-34.5%), respectively. Pregnancy in adolescence appears to constitute a psychosocial problem rather than a biological risk. PMID- 11049095 TI - Prevalence of Haemophilus influenzae pharyngeal carriers in the school population of Catalonia. Working Group on invasive disease caused by Haemophilus influenzae. AB - The objective of this study was to determine the prevalence of healthy Haemophilus influenzae (Hi) pharyngeal carriers in a representative sample of the Catalonian school population, as well as the factors associated. A two-stage cluster sampling was carried out. Parents were given a questionnaire to collect information on sociodemographic and epidemiological variables. A pharyngeal swab was performed on children when informed consent was given by parents, and was cultured on chocolate agar with 260 microg/ml bacitracin. Of the 1212 children studied, 316 (26%) H. influenzae carriers were detected: 5 (0.4%) serotype b, 1 (0.08%) serotype c, 6 (0.5%) serotype e, 5 (0.4%) serotype f, and 299 (24.7%) non typable. Age, gender and geographical location were the only variables associated with H. influenzae carrier status. The prevalence of non-typable H. influenzae carriers was similar to that of studies carried out in other countries, while that of serotype b carriers was similar to the remainder of H. influenzae capsulates, and lower than that described in previous studies. These data are in accordance with the low incidence of the disease observed in our context, although the possibility that the vaccine coverage may have affected the results of this study cannot be dismissed. PMID- 11049097 TI - Validity of the hospital discharge diagnosis in epidemiologic studies of biliopancreatic pathology. PANKRAS II Study Group. AB - BACKGROUND: The aim was to analyse the magnitude, direction and predictors of change in the main hospital discharge diagnosis (HDD) after a clinical expert review, among patients included in a multicentre molecular epidemiologic study of biliopancreatic diseases. METHODS: A total of 602 patients with a suspicion diagnosis of pancreas cancer (PC), cancer of the extrahepatic biliary system (CEBS) or benign biliopancreatic pathologies (BPP) were prospectively recruited at five general hospitals. A structured form was used to collect information from medical records. A panel of experts revised all diagnostic information and established the main clinicopathological diagnosis (CPD) by consensus. RESULTS: Of the 204 cases with a HDD of PC, 176 (86%) were deemed to have a CPD of PC, eight of CEBS, twelve a neoplasm of different origin, four BPP and four syndromic diagnoses. Thus, 28 cases (14%) were false positives. Of the 129 patients with a HDD of CEBS, 15 (12%) were false positives. Nine of the 396 cases with a HDD of non-PC (2%) had a CPD of PC (false negatives), whilst 14 of 471 patients with a HDD of non-CEBS (3%) were deemed to have CEBS. Overall, sensitivity and specificity of HDD for PC were, respectively, 95 and 93%, and for CEBS, 89 and 97%. Cytohistological confirmation and laparotomy were independent predictors of diagnostic change. CONCLUSIONS: Validity of the HDD was high, but its association with some clinical variables suggests that sole reliance on HDD can significantly bias results, and highlights the need to review all HDDs. Alternatively, only patients at high risk of misdiagnosis could be reviewed: primarily, those lacking a cytohistological diagnosis or a laparotomy. No exclusions appear warranted solely on the basis of age, gender or tumour spread. PMID- 11049098 TI - Sequential logistic models for 30 days mortality after CABG: pre-operative, intra operative and post-operative experience--The Israeli CABG study (ISCAB). Three models for early mortality after CABG. AB - OBJECTIVES: The goal of this paper was to examine the added effect of operative and post-operative variables on 30 days mortality, in addition to patients' case mix factors. SETTING AND DESIGN: A prospective study of 4835 patients, 95% of all Israeli patients who underwent coronary artery bypass grafting (CABG) in 1994. Information related to risk of death was collected at admission to hospital (preceding the operation), at time of the operation and in the immediate post operative period. Deaths were independently ascertained. METHOD: Data collectors followed every patient from admission to discharge. Sequential logistic models were constructed for the 'case-mix', 'operative' and the 'post-operative' periods in chronological order. Each model incorporated and adjusted for the risk estimated at the previous point in time, by forcing individual risk scores. RESULTS: Significant pre-operative risk factors for 30 days mortality, in the case-mix model included mainly severity of illness characteristics, such as, left ventricular dysfunction and emergency admission, (c-statistic 78.8%). Model 2 (the 'operation' model) included in addition to the case-mix score, excessive duration of the operation per graft, bleeding, etc. (c-statistic 85.3%). The post operative model showed the added effect of the post-operative factors such as low haemoglobin, additional surgery, and excessive time on respirator, (c-statistic 92.4%). CONCLUSIONS: The sequential analysis was an efficient method for updating patients' risk over time, where the number of events was small, relative to the number of risk factors. The addition of peri-operative factors increased significantly the predictive power of the model, adding clinical insights to the role of the hospital experience on 30 days mortality. PMID- 11049099 TI - Definitions of healthy eating in Spain as compared to other European Member States. AB - STUDY OBJECTIVE: To assess what healthy eating means for the European population and whether this concept differs between Spain and other European Countries. DESIGN: A Pan-European survey was developed between October 1995 and February 1996 by the Institute of European Food Studies (Dublin). Each subject was asked to describe in his or her own words what he/she understood by 'healthy eating'. Comparisons were made among four groups of European countries (Northern, Central, Spain, and other Mediterranean countries). SETTING: The survey included participants from the 15 member states of the European Union, selecting quota controlled samples to make them nationally representative. SUBJECTS: The questionnaire was completed by 14,331 persons, approximately 1000 from each country. RESULTS: The responses were grouped into 89 broad categories of similar answers concerning nutritional value and afterwards these responses were collapsed to simplify the presentation. The definition of healthy eating such as 'more fiber' and 'less fat' was more prevalent in other States, members of the European Union than in Mediterranean Countries, although the definition of 'balanced diet' was more frequently mentioned in Spain than in the rest of the European Union. CONCLUSIONS: Our results show that the concept of 'balance and variety' is more prevalent in Spaniards than in other traditional Mediterranean countries. Differences in the definitions of healthy eating among European countries could be explained, at least partially by differences in consumption patterns and in the nutrition education. PMID- 11049100 TI - Role of socioeconomic indicators in the prediction of all causes and coronary heart disease mortality in over 12,000 men--The Italian RIFLE pooling project. AB - The relationship of socioeconomic indicators (education, occupation and residence) to short-term all cause mortality and coronary heart disease (CHD) mortality was evaluated in an Italian population sample. Socioeconomic indicators (education, occupational level and residence) and major CHD risk factors were measured in 12,361 males aged 40-69 years; mortality data by cause were collected for the next 6 years. All cause and CHD mortality risk ratio (RR) in the different educational and occupational levels and residence were computed by Cox proportional hazards models. After 6 years 385 men died, of whom 105 were coronary fatalities. No association with educational level was found for all cause mortality (RR: 1.00 high, 0.71 medium, 0.77 low) and for CHD mortality (RR: 1.00 high, 0.39 intermediate, 0.71 low). Occupational level was significantly associated (p < 0.031) with all cause mortality (RR: 1.00 high-intermediate, 1.27 low). Urban vs. rural residence (RR: 1.00) showed a RR for all cause mortality of 1.33 (p < 0.011). Adjustment for bio-behavioral risk factors did not change the above results; only mortality for CHD of urban vs. rural residents increased (RR: 1.94, p = 0.004). In conclusion the negative association of mortality with occupational level, albeit not with education, indicates that occupation is a better indicator of socioeconomic status in Italy. Status incongruity as well as residence in an urban environment could be risk conditions for total and CHD mortality. PMID- 11049101 TI - Modeling the HIV/AIDS epidemic via survivor functions. AB - An original approach to simulation modeling of the HIV/AIDS epidemic is proposed. This approach uses survivor functions estimated from cohort studies conducted with seropositive and AIDS-diagnosed individuals. The model can be considered an alternative to the usual Markov models and accounts for time-dependent HIV progression to AIDS, and AIDS progression to death. By using various forms of survivor functions, it can also easily be extended to accommodate natural history events, as well as long-term survivors and cofactor effects, when appropriate data are available. PMID- 11049102 TI - Epidemiological aspects of brucellosis in Jordan. AB - From January 1988 to December 1997, a total of 7842 cases of human brucellosis were registered at the Ministry of Health in Jordan. A link was found to exist between the lambing season and the occurrence of the infection. The number of cases was found to be the lowest in children below 4 years and highest in the 5 14-years age group. Incidence of the infection was calculated per 100,000 population. The lowest incidence of brucellosis was 16.7 and this was detected in the year 1988, whereas the highest was 29.9 and this was observed in 1991. Evidence is provided which indicates that notified cases of human brucellosis in Jordan do not reflect the actual frequency but rather underestimate the extent of the infection. PMID- 11049103 TI - Varying incorporation of fatty acids into phospholipids from muscle, adipose and pancreatic exocrine tissues and thymocytes in adult rats fed with diets rich in different fatty acids. AB - Despite numerous studies, the importance which the tissue or the composition of the diet may have in the biological distribution of each fatty acid is not well known. To determine the importance of tissue origin and dietary fatty acids in the fatty acid composition of cell phospholipids, 54 male adult rats were fed isocaloric diets for one month varying only in their fatty acid compositions. The fat component of the six experimental groups was derived from olive oil, sunflower oil, fish oil, soybean oil, palmitic acid, or 82% palmitic acid plus 18% soybean oil, supplying the essential fatty acid. The fatty acid composition of phospholipids from thymocytes, pancreatic exocrine, muscle and adipose tissues was studied by gas-chromatography. The tissue of origin was a more important source of variation than diet in the fatty acid content of the cell phospholipids except for palmitic acid (16:0), eicosapentaenoic acid (20:5 n-3), and docosahexaenoic acid (22:6 n-3). This study points out the complexity of the interrelations between different families of fatty acids and of the specificity of each tissue to changes in the composition of dietary fatty acids, as well as the inconvenience of speaking from the dietary point of view of groups of fatty acid families based on the position of the double bond, since their individual behaviour, including saturated fatty acids, is very different in the face of dietary manipulation. The study also highlights the different behaviour of each of the fatty acids in relation to the others in the diet in each of the tissues, a circumstance which should be taken into account when evaluating the biological effects in both epidemiological and experimental studies. PMID- 11049104 TI - Decreased organ failure in patients with severe SIRS and septic shock treated with the platelet-activating factor antagonist TCV-309: a prospective, multicenter, double-blind, randomized phase II trial. TCV-309 Septic Shock Study Group. AB - Sepsis and organ failure remain the main cause of death on the ICU. Sepsis is characterized by a severe inflammatory response, in which platelet-activating factor (PAF) is considered to play an important role. This study investigated whether treatment with the PAF-antagonist TCV-309 reduces morbidity and mortality in patients with septic shock. The study was conducted as a double-blind, randomized, placebo controlled multicenter study. The included patients had to fulfill the SIRS criteria with a clinical suspicion of infection, an admission APACHE II score greater than 15, and shock, defined as a mean arterial pressure <70 mmHg and/or a decrease > or =40 mmHg despite adequate fluid resuscitation. Patients received 1.0 mg/kg TCV-309 or placebo, twice daily, intravenously during 14 days. The prospectively set goals were MOF score, recovery from shock, mortality, and assessment of the safety of the medication. A total of 98 patients were included of which 97 were analyzed on an intention-to-treat basis. The overall survival at day 56 of TCV-309 treated patients was similar compared to placebo treated patients (51.0% vs. 41.7%, P = 0.47). In contrast, the mean percentage of failed organs per patient present after 14 days in the TCV-309 treated patients was significantly lower compared to the placebo treated patients (11.9% vs. 25.1%, P = 0.04), leading to a reduced need for vasopressors, dialysis, and ventilatory support. Furthermore, the mean APACHE-II score during treatment with TCV-309 was significantly lower and the number of patients recovered from shock after day 14 was significantly higher in the TCV-309 treated patient group (2/32 vs. 9/29, P = 0.01). The number of adverse events was not significantly different between the TCV-309 and placebo treated patients. TCV-309 did not change overall mortality of septic shock, however a substantial reduction in organ dysfunction and morbidity, frequently associated with septic shock was achieved, without significant adverse events. PMID- 11049105 TI - Ischemic preconditioning ameliorates ischemia- and reperfusion-induced intestinal epithelial hyperpermeability in rats. AB - We hypothesized that ischemic preconditioning (IPC) would ameliorate ischemia (I) and reperfusion (R)-induced intestinal mucosal hyperpermeability and that this effect would be diminished by lowering local adenosine concentrations using adenosine deaminase (ADA). The small intestine of anesthetized rats (group 1; n = 6) was divided into six 10-cm segments (A1-F1) each perfused by a different set of mesenteric branches. Segments D1-F1 were subjected to 3 cycles of IPC (2 min I/5 min R). Segments A1, B1, and C1 were excised at baseline, after 60 min of I (160), and after 60 min of I followed by 60 min of R (160/R60), respectively. Segment D1 was excised immediately after the last cycle of IPC, E1 was excised at 160 after IPC, and F1 was excised at 160/R60 after IPC. In group 2 (n = 6), the intestine was divided into five 10-cm vascularly isolated segments (A2-E2). Segment A2 was resected at baseline. The lumen of the remaining segments was filled with ADA (32 U/50 cm). Segment B2 was removed at the end of the experiment having been exposed to ADA for 150 min (ADA150). Segments C2, D2, and E2 were subjected to IPC. Segment C2 was excised immediately thereafter. Segments D2 and E2 were excised at 160 and 160/R60, respectively. Intestinal permeability to fluorescein isothiocyanate-labeled dextran (molecular weight 4000 D) was assessed ex vivo by using an everted gut sac method. IPC ameliorated intestinal hyperpermeability induced by 160 (43.0+/-7.6 vs. 70.4+/-8.3 nLmin/cm2; P = 0.024) and 160/R60 (20.2+/-3.7 vs. 69.5+/-10.8 nL/min/cm2; P= 0.003). IPC prevented ischemia-induced reduction in villus height. Treatment with ADA partially reversed the protective effect of IPC on the changes in permeability and villus height induced by I/R. We conclude that IPC partially protects against mucosal barrier dysfunction in rats subjected to mesenteric I/R. Adenosine is a mediator of IPC in the gut mucosa, but other factors also may be important. PMID- 11049106 TI - Adherence regulates macrophage signal transduction and primes tumor necrosis factor production. AB - While monocyte/macrophage (Mphi) adherence to a matrix is necessary for differentiation and prolonged survival, the effect of adherence on the signaling mechanisms responsible for Mphi activation is unknown. Lipopolysaccharide (LPS) activates Mphi by signaling through members of the mitogen activated protein kinase (MAPK) family thereby inducing transcription of proinflammatory cytokines, such as TNF-alpha. Since adherence has been shown to affect different activities of various myeloid phagocytes, we investigated whether adherence affects intracellular signaling and modulates activation of the Mphi proinflammatory phenotype. We assessed the effect of adherence on activation of rabbit alveolar Mphi by measuring LPS-induced TNF-alpha mRNA and TNF-alpha secreted product in adherent versus nonadherent cells, in vitro. The effect of adherence on LPS induced activation of MAPK was assessed by western analysis using a dual phosphospecific antibody against p38MAPK, p42,44ERK, and p54SAPK. LPS is known to induce activation of NF-kappaB and AP-1. Modulation of these two transcription factors by LPS under adherent versus nonadherent conditions was evaluated by gel shift analyses. The results were that adherent cells treated with LPS, 10 ng/mL or 1 microg/ml, elicited a 26- and 132-fold increase, respectively, in TNF-alpha production. Nonadherent cells did not elicit significant TNF-alpha in response to LPS. Adherence alone induced significant ERK and AP-1 activation, but did not stimulate a significant TNF-alpha response and no further activation of ERK and AP-1 was observed with LPS stimulation. Adherence alone did not activate p38MAPK or NF-kappaB, but primed Mphi for an augmented response to LPS in activation of p38, NF-kappaB and in production of TNF-alpha. We conclude that adherence primes Mphi for activation and regulates MAPK signal transduction pathways. PMID- 11049107 TI - Central versus peripheral mediation of naloxone's perfusion effects in endotoxic rats. AB - Opioid receptor antagonists can act centrally and peripherally. It is unclear if these 2 pathways differentially mediate the perfusion-associated effects of opioid antagonism during endotoxemia. Male, Sprague-Dawley rats (340-390 g) were surgically prepared with left ventricular, tail artery, and jugular vein catheters 24 h before experiments were begun. Conscious, unrestrained rats were challenged with Escherichia coli lipopolysaccharide (LPS; 2 mg/kg/hr over 30 min) infusion. Measurements of regional blood flows were made using radioactive microspheres prior to (baseline), and at 60 and 120 min after LPS infusion. Saline (1 mL/kg bolus + 0.5 mL/kg/h infusion), naloxone (Nlx; 4 mg/kg bolus + 2 mg/kg/h infusion), or naloxone methyl bromide (Nlx-mb; 4.64 mg/kg, bolus + 2.32 mg/kg/h infusion) were administered 40 min after LPS infusion was begun. Nlx-mb does not cross the blood-brain barrier, and was thus used to differentiate central from peripherally mediated responses. At the end of each experiment, blood samples were collected for determination of ET-1 and nitric oxide metabolites (NOx = NO3 + NO2) using enzyme-linked immunosorbent assay (ELISA) and Griess reaction methods, respectively. Endotoxemia produced a significant decrease in cardiac output and an increase in systemic vascular resistance. Treatment with Nlx or Nlx-mb significantly attenuated the endotoxin-induced elevation in systemic vascular resistance and the decrease in cardiac output at 60 min after induction of endotoxemia compared with their respective baseline values. Nlx and Nlx-mb also attenuated the endotoxin-induced increases in hepatic portal and skeletal vascular resistances. These observations suggested that the ameliorative effect of Nlx on endotoxemia-induced regional vascular resistance alterations was mediated via peripheral opioid receptor mechanisms. However, although Nlx attenuated the endotoxin-induced decreases in the blood flow to the stomach and pancreas, Nlx-mb attenuated the endotoxin-induced decreases in the blood flow to the small intestine and cecum, in addition to the pancreas and, to some extent, the stomach. As such, separate central and peripherally mediated actions of opioid receptor antagonism were indicated. Nlx also resulted in an increase in the plasma levels of ET-1 only, whereas Nlx-mb increased the plasma levels of ET-1 and NOx. These observations suggest that separate central and peripheral effects of opioids during endotoxemia play a role in the associated circulatory alterations, and may differentially affect the release and/or synthesis of vasoactive mediators that might be related to their varied hepatosplanchnic vascular response during endotoxemia. PMID- 11049108 TI - Heat shock inhibits phosphorylation of I-kappaBalpha. AB - Previous studies demonstrated that induction of the heat shock response is associated with inhibition of the proinflammatory transcription factor NF-kappaB by a mechanism involving inhibition of I-kappaBalpha degradation. To provide further insight regarding the interactions of these fundamental cellular responses, the present experiments were designed to elucidate the mechanism(s) by which heat shock inhibits degradation of I-kappaBalpha. In an in vitro model of inflammatory cell signaling, treatment of RAW 264.7 murine macrophages with LPS (100 ng/mL) caused rapid degradation of I-kappaBalpha. Heat shock, 1 h before treatment with LPS, completely inhibited LPS-mediated degradation of I kappaBalpha. Immunoprecipitation studies demonstrated that heat shock inhibited LPS-mediated ubiquitination of I-kappaBalpha. Western-blot analyses using a phosphorylated I-kappaBalpha-specific antibody demonstrated that heat shock inhibited LPS-mediated phosphorylation of I-kappaBalpha. In contrast, heat shock induced phosphorylation of c-jun. In murine fibroblasts having genetic ablation of the heat shock factor-1 gene, heat shock inhibited tumor necrosis factor-alpha mediated degradation of I-kappaBalpha. We conclude that the mechanism by which heat shock inhibits LPS-mediated degradation of I-kappaBalpha involves specific inhibition of I-kappaBalpha phosphorylation and subsequent I-kappaBalpha ubiquitination. In addition, this mechanism does not involve activation of heat shock factor-1 or the heat shock proteins regulated by heat shock factor-1. PMID- 11049109 TI - Hemodynamic pathogenesis of ischemic hepatic injury following cardiogenic shock/resuscitation. AB - Post-ischemic hepatic injury is observed commonly following cardiogenic or hypovolemic shock. We evaluated the putative roles of the alpha-adrenergic sympathetic nervous system and the renin-angiotensin axis in the pathogenesis of hepatic injury following cardiogenic shock. Previous studies have characterized the hepatic hemodynamic response to shock, while the relationship of these hemodynamic changes to ischemic hepatic injury has not been defined. Sustained (4 h) periods of pericardial tamponade (after mild hemorrhage) followed by 2 h of resuscitation generated a reproducible model of cardiogenic shock and consequent post-ischemic hepatic injury in anesthetized pigs. In a separate group of pigs, the alpha-adrenergic component of the sympathetic nervous system was ablated with phenoxybenzamine or, in other groups, the renin-angiotensin axis was ablated by either prior nephrectomy or, separately, by confirmed angiotensin converting enzyme inhibition with teprotide. The hepatic injury response in each case was reevaluated. Compared to sham-shocked pigs, those subjected to tamponade alone manifested selective splanchnic vasospasm and consequent biochemical and histological evidence of classic post-ischemic liver injury (centrilobular necrosis involving about a third of each hepatic lobule). These manifestations of splanchnic vasospasm and the consequent ischemic injury were not ameliorated by confirmed alpha-adrenergic blockade, but significantly attenuated by either method of prior ablation of the renin-angiotensin axis. This model of sustained cardiogenic shock and resuscitation generates the manifestations of ischemic hepatic injury associated with selective splanchnic vasospasm, findings consistent with previous, short-term, hemodynamic studies. The major mediator of this response, and the consequent hepatic injury, is the selective hypersensitivity of the mesenteric vasculature to the renin-angiotensin axis. PMID- 11049111 TI - Androgen and estrogen receptors in splenic T lymphocytes: effects of flutamide and trauma-hemorrhage. AB - The endogenous sex steroids, testosterone and beta-estradiol, play a major role in inflammatory processes. They regulate several cytokine genes by interaction with their intracellular receptors that are, essentially, transcription factors. Because T-lymphocyte functions are altered following trauma-hemorrhage in male mice, we investigated whether (i) receptors for androgen (AR) and estrogen (ER) are present in splenic T lymphocytes, (ii) receptor expressions are altered following trauma-hemorrhage, and (iii) pretreatment of male mice with the AR antagonist, flutamide, alters receptor expressions and IL-6 release. Analysis of nuclear extracts indicated the presence of AR and ER in splenic T lymphocytes. No difference in receptor expressions between males and females or following trauma hemorrhage was observed. Pretreatment of males with flutamide, however, led to increased ER expression in T lymphocytes of sham and trauma-hemorrhaged animals. This suggested that flutamide is capable of inducing the expression of another receptor belonging to a different gonadal steroid. Because response elements for AR and ER are present in the promoter region of the IL-6 gene, release of IL-6 and expression of signal transducer and activator of transcription 3 (STAT3) were analyzed as functional parameters in splenic T lymphocytes. Trauma-hemorrhage decreased IL-6 release by T lymphocytes and the release was restored to sham levels with flutamide pre-treatment. Similarly, STAT3 expression was decreased in T lymphocytes following trauma-hemorrhage and the expression was restored by flutamide pre-treatment. These data collectively demonstrate the importance of gonadal steroids in the regulation of splenic T-lymphocyte functions. PMID- 11049110 TI - Injection of iron compounds followed by induction of the stress response causes tissue injury and apoptosis. AB - To determine whether iron-laden tissue subsequently stimulated to produce the stress ("heat shock") response-sustained injury, hindlimbs of male ND4 mice were injected with iron salts, hemin, or hemoglobin. The stress response was induced with sodium arsenite or with heat. Ulcers appeared at the injection site. Tissues were analyzed by three distinct techniques-electron microscopy, TUNEL stain, and agarose gel electrophoresis of low molecular weight DNA-which collectively suggest that the tissue injury is, at least in part, the consequence of accelerated apoptosis. The data suggest that the toxicity of free iron is amplified by induction of the stress (heat shock) response to signal a programmed response. This model and mechanism may have implications in pathological processes ranging from the cutaneous wounds of venous stasis disease to the tissue failure of multiple organ dysfunction. PMID- 11049112 TI - Angiotensin II blockade in existing hypovolemia: effects of candesartan in the porcine splanchnic and renal circulation. AB - Angiotensin II (AngII) is an important vasoconstrictor during hypovolemia. This study focused on the effects of the AngII receptor blocker candesartan on intestinal, hepatic, and renal hemodynamics during severe hypovolemia when administered in preexisting moderate hypovolemia. It was hypothesized that specific AngII receptor blockade might enhance splanchnic perfusion during hypovolemia. Fasted, anesthetized, ventilated, juvenile pigs were hemorrhaged by 20% of the blood volume for 30 min. Animals were then randomized to receive candesartan (CAND, n = 11) or the vehicle (CTRL, n = 10) prior to further hemorrhage to 40% of the blood volume for 30 min. The shed blood was then retransfused. Systemic and splanchnic hemodynamics were recorded including intestinal mucosal, superficial and parenchymal hepatic, and cortical and medullary renal microcirculation by laser-Doppler flowmetry. Arterial blood gases were analysed. Candesartan-treated animals maintained mesenteric and jejunal mucosal perfusion during 40% hypovolemia compared to CTRL animals, while no differences were observed in the hepatic and renal circulation. Retransfusion restored mesenteric and renal blood flows despite persistent hypotension and reduced cardiac output in both CAND and CTRL animals. Renal medullary and hepatic parenchymal microcirculation failed to recover during retransfusion in both CAND and CTRL animals. Arterial acidosis, hypercarbia, and a negative base excess were observed in CTRL animals following retransfusion whereas those parameters were normalised in CAND animals. Administration of candesartan in moderate hypovolemia ameliorated the reduction and consequences of mesenteric and intestinal, but not hepatic perfusion during severe hypovolemia. No adverse effects were observed in the renal circulation. PMID- 11049113 TI - Lipopolysaccharide causes atrophy of Peyer's patches and an increased expression of CD28 and B7 costimulatory ligands. AB - Intestinal mucosal dysfunction appears to contribute to infectious complications in critically ill patients. The current study was undertaken to investigate whether endotoxin affects lymphocyte subpopulations and the expression of costimulatory signals in Peyer's patches (PP). Female Balb/c mice were given an intraperitoneal injection of 25 microg LPS and sacrified 24 h or 72 h later to determine total cell yield, lymphocyte subpopulations (B-cells, total T-cells, CD4+- and CD8+-cells), the costimulatory molecules CD28, B7.1 (CD80) and B7.2 (CD86) and the percentage of apoptotic cells in PP and in the spleen as well as small intestinal IgA concentration. Lipopolysaccharide (LPS) challenge caused a significant decrease of total cell yield in PP at both time-points (-50+/-28% and -43+/-25%, respectively; P < 0.001). This decrease was significant for all measured lymphocyte subpopulations. In contrast, total cell yield was increased (P < 0.001) in the spleen 24 h (+52+/-13%) and 72 h (+130+/-22%) after LPS. The decrease of lymphocyte numbers in the PP was accompanied by an increased percentage of lymphocytes expressing costimulatory molecules. In this respect, an increased percentage of CD40+CD80+, CD40+CD86+, and of CD4+CD28+ could be demonstrated after LPS administration. In the spleen, the percentage of CD4+CD28+ was also elevated after LPS bolus, however, the percentage of CD40+CD80+ was reduced, and that of CD40+CD86+ was unaltered. The influence of LPS on apoptosis of lymphocytes was time-dependent. The percentage of apoptotic cells 24 h after LPS was increased in PP (P < 0.01), but was unchanged in the spleen. Seventy-two hours after LPS injection, the percentage of apoptotic cells returned to normal in PP. Luminal IgA levels remained unchanged after LPS challenge. In conclusion, our data show that LPS causes atrophy of PP which seems to be counterregulated by an enhanced expression of costimulatory molecules. PMID- 11049115 TI - Platelet-activating factor (PAF)-induced decreases in whole-body and skeletal muscle protein synthesis. AB - Platelet-activating factor (PAF) has been shown to reduce rat skeletal muscle amino acid uptake, which may restrict intracellular amino acid availability for protein synthesis and amino acid oxidation during endotoxemia. We investigated in rats the effect of PAF infusion on amino acid and protein metabolism by measuring (a) whole-body and tissue leucine kinetics; (b) plasma amino acid profile; and (c) muscle RNA activity (protein synthesis efficiency) and relative abundance of myofibrillar proteins. Fasted male Sprague-Dawley rats (250+/-20 g) were given a 4-h i.v. continuous infusion of L-(1-14C)-leucine to determine leucine kinetics during the infusion of PAF (2 microg/kg PAF as a priming i.v. bolus 1 h before a 4-h i.v. infusion of 2 microg/kg/h PAF) or vehicle. PAF infusion caused sustained hypotension, hyperglycemia, hematological alterations, and hyperlacticacidemia. Whole-body protein synthesis was decreased by 24% (P < 0.05) and leucine flux oxidized was increased by 23% (P < 0.05). Leucine flux was reduced, although not significantly (P = 0.07), in PAF-treated rats (n = 8) compared with controls (n = 8). PAF significantly decreased fractional protein synthesis in the rectus abdominus (33%), soleus (30%), and extensor digitorum longus (26%) muscles, but not in the liver. Plasma branched-chain amino acid levels decreased (approximately 30%, P < 0.05) in PAF-treated rats. Muscle RNA activity was 32% lower and myosin relative abundance declined whereas actin was unchanged in PAF treated rats. PAF induced net protein catabolism as a result of elevated leucine oxidation at the expense of protein synthesis. PAF had the cumulative effects in the skeletal muscle of (a) attenuating amino acid uptake, (b) reducing protein synthesis efficiency, (c) decreasing fractional protein synthesis rate, and (d) decreasing myosin relative abundance. Thus, PAF may be an important mediator of decreased protein synthesis in skeletal muscle during endotoxic and septic shock. PMID- 11049114 TI - Plasma concentration of biologically active fibronectin and fibronectin bound to gelatin-like material in a porcine model of hyperdynamic endotoxic shock. AB - Due to its opsonizing role, plasma fibronectin (pFN) binds to circulating degradation products deriving from collagenous structures and mediates their elimination by the reticuloendothelial system (RES). In septic shock, an overflow of this material may lead to a lack of pFN and an impaired RES activity. In fact, low pFN levels have been reported to correlate with unfavorable clinical outcome. However, dysfunction of the RES is also caused by other shock related factors, and death from septic shock also has been observed in the presence of normal FN levels. To investigate the involvement of opsonic FN in the progression of sepsis, we discriminated between biologically active FN and FN bound to gelatin like material in pigs developing a hyperdynamic endotoxic shock. All FN determinations were performed with the immunochemical assay. Discrimination between free FN and complexed FN was achieved by separation on gelatin sepharose. A continuous decrease of total FN and free FN was observed in the septic group reaching 57% and 50% of the initial level at the end of the 5-h observation period, respectively. However, a significant difference was not detected before both the microcirculatory and macrocirculatory alterations indicative of hyperdynamic endotoxic shock were completely established. Complexed FN was increased slightly in both groups without any group specific differences. We conclude that the FN-mediated opsonization of circulating gelatine-like material does not play a critical role in early circulatory shock. PMID- 11049116 TI - Are the effects of platelet-activating factor on muscle metabolism of carbohydrates, amino acids and proteins attributable mainly to epinephrine? PMID- 11049117 TI - Somatic growth following the modified Fontan procedure. PMID- 11049118 TI - Improvement in growth, and levels of insulin-like growth factor-I in the serum, after cavopulmonary connections. AB - INTRODUCTION: The total cavopulmonary connection, and the bidirectional Glenn anastomosis, are widely used for palliation of patients with complex functionally univentricular hearts. Little attention has been paid to the potential for postoperative growth in children after these operations, which are now performed at increasingly younger age. MATERIAL AND RESULTS: Physical growth, and levels of insulin-like growth factor I in the serum, were measured in 20 patients, aged 11.5 +/- 5.6 years, 2 (0.5-6) years after a total cavopulmonary connection in 12, or a Glenn anastomosis in 8. All patients were in functional class I or II of the categorisation of the New York Heart Association, with excellent haemodynamic and angiographic findings. None of the patients had clinical signs of protein losing enteropathy. Controls included 33 healthy children, aged 11.5 +/- 2.7 years. Preoperatively, the mean Z-scores for weight and height were negative, -1.1 +/- 0.8 and -0.5 +/- 1.5. At follow-up, both parameters had improved significantly by 1.1 +/- 0.9 and 0.8 +/- 1.2 percentiles, and Z-scores were comparable between the two groups (p=0.81 for weight and p=0.88 for height). No correlations were found between haemodynamics and the improvement in growth noted during follow-up. Increases equal to, or greater than 2 standard deviations for weight and height were seen only in children undergoing surgery before the age of 5 years. A significant correlation between age at operation and improvement in growth, however, could not be found. Levels of growth factor measured in the serum were not statistically different from levels in healthy children for either group of patients (p=0.07 for girls and p=0.37 for boys). CONCLUSION: Physical growth improved significantly following the surgical procedures. The concentrations of the growth factor measured in the serum were not different from levels in healthy children, suggesting normal nutritional status in both palliative situations. PMID- 11049120 TI - Should we standardise the pre-operative management of babies with complete transposition? AB - BACKGROUND: Complete transposition is the most common form of neonatal cyanotic heart disease. The management of this condition has changed markedly in the last decade and there appears to be a significant variation between centres in terms of pre-operative management. OBJECTIVES/METHODS: We surveyed all paediatric cardiac surgical centres in the United Kingdom regarding pre-operative management, particularly performance and timing of balloon atrial septostomy and aortogram, imaging techniques used and discharge prior to surgery. RESULTS: There is significant variation in pre-operative management: 10 centres now perform septostomy outside the catheter lab and 11 without general anaesthesia. Eight centres use echo control only and only 3 perform routine aortograms. Three centres do not perform routine septostomy. The most common age for arterial switch was at 1-2 weeks, but some routinely performed this procedure up to 1 month of age and others aim for arterial switch before one week of age. Only 3 centres routinely discharge patients between septostomy and switch. CONCLUSIONS: Despite a trend towards echo guided septostomy and earlier arterial switch there is still considerable variation in early management of patients with transposition of the great arteries. Debate within the profession leading to a more standardised pre-operative management strategy would protect both the patient and the doctor involved in the care of children with complete transposition. PMID- 11049121 TI - Transcatheter closure of atrial septal defects demands co-operation between the interventionist and the echocardiographer. PMID- 11049119 TI - Somatic growth failure after the Fontan operation. AB - Our study was designed to characterize the patterns of growth, in the medium term, of children with functionally univentricular hearts managed with a hemi Fontan procedure in infancy, followed by a modified Fontan operation in early childhood. Failure of growth is common in patients with congenital cardiac malformations, and may be related to congestive heart failure and hypoxia. Repair of simple lesions appears to reverse the retardation in growth. Palliation of the functionally single ventricular physiology with a staged Fontan operation reduces the adverse effects of hypoxemia and prolonged ventricular volume overload. The impact of this approach on somatic growth is unknown. Retrospectively, we reviewed the parameters of growth of all children with functionally univentricular hearts followed primarily at our institution who had completed a staged construction of the Fontan circulation between January 1990 and December 1995. Measurements were available on all children prior to surgery, and annually for three years following the Fontan operation. Data was obtained on siblings and parents for comparative purposes. The criterions of eligibility for inclusion were satisfied by 65 patients. The mean Z score for weight was -1.5 +/- 1.2 at the time of the hemi-Fontan operation. Weight improved by the time of completion of the Fontan circulation (-0.91 +/- 0.99), and for the first two years following the Fontan operation, but never normalized. The mean Z scores for height at the hemi-Fontan and Fontan operations were -0.67 +/- 1.1 and -0.89 +/- 1.2 respectively. At most recent follow-up, with a mean age of 6.1 +/- 1.3 years, and a mean time from the Fontan operation of 4.4 +/- 1.4 years, the mean Z score for height was -1.15 +/- 1.2, and was significantly less than comparable Z scores for parents and siblings. In our experience, children with functionally univentricular hearts who have been palliated with a Fontan operation are significantly underweight and shorter than the general population and their siblings. PMID- 11049122 TI - The anatomy of interatrial communications--what does the interventionist need to know? AB - Increasingly, the interventional cardiologist is seeking to close interatrial communications by inserting devices by means of catheterisation. So as to optimise these procedures, it is advantageous to have a firm grasp of the anatomy of the normal atrial septal structures, this then providing the basis to understand the morphology of the holes which can exist between the chambers, not all of which are true septal defects. A true septal structure can be removed without exiting from the cavities of the heart. It is the flap valve of the oval fossa, along with the anterior rim of the fossa, which fulfill this criterion. The remainder of the extensive rim of the normal fossa is no more than an infolding between the walls of the right and left atria and their venous tributaries, and has different dimensions at various points around the circumference. The so-called muscular atrioventricular "septum" is a sandwich incorporating a layer of epicardial fibro-adipose tissue. True defects of the atrial septum, therefore, exist because of deficiency, perforation, or absence of the flap valve. Most of these defects will prove suitable for interventional closure, but potential caveats include multiple defects, aneurysm of the flap valve, or adjacency of the fossa to the venous orifices. The other interatrial communications, namely the sinus venosus, coronary sinus, and "ostium primum" defects are outside the confines of the oval fossa. Recognition of this feature is the key to their diagnosis, and their differentiation from true atrial septal defects. Of these defects, only the coronary sinus defect is likely to be suitable for device closure, and then only in the very rare circumstances when it is seen in isolation. PMID- 11049123 TI - The role of echocardiography in transcatheter closure of atrial septal defects. AB - Closure of so-called "secundum" atrial septal defects with a device inserted on a catheter necessitates precise delineation of their morphology. Echocardiography is the diagnostic method of choice to demonstrate this morphology, and to differentiate such defects located within the oval fossa from the other variants producing an interatrial communication. Precordial echocardiography usually allows selection of cases likely to be suitable for closure in this fashion. This selection is based on the localisation and the size of the deficiency in the oval fossa, the length of the interatrial septum, and the adequacy of the infolded rims surrounding the defect. Suitability for closure is reevaluated by transesophageal echocardiography, either as a separate investigation or at the start of the interventional catheterisation. This investigation requires a multiplane transesophageal echocardiographic probe, since only oblique planes will demonstrate the entrance of the systemic and pulmonary veins and their relationship to the defect. Transesophageal echocardiography serves as an important monitoring tool during the interventional procedure. As such, it is a necessary adjunct to fluoroscopy. The stretched diameter of the defect measured with a balloon is the main determinant of the choice of the type and size of the device. This diameter can be measured fluoroscopically, as well as on echo. Colorflow mapping serves to rule out residual shunting during the occlusion of the defect with the balloon. During deployment of the device, constant echocardiographic visualisation of the device and its position relative to the atrial septum facilitates proper placement. Such constant visualisation can only be provided by repeated quick acquisitions of multiple planes. Once the device is released, the investigator should continue to record the position of the device, and assess the potential for residual shunting. Most of the devices show some subtle change in position during the first 20 minutes after implantation. PMID- 11049124 TI - Three-dimensional echocardiography in transcatheter closure of atrial septal defects. PMID- 11049125 TI - The Amplatzer septal occluder. AB - The Amplatzer Septal Occluder is made from a Nitinol wire mesh shaped into 2 disks with a connecting waist, which serves to center the device in the defect while occluding it. The Amplatzer device is also available in a configuration with no central waist for use in patients with patent oval foramen, or multi perforated aneurysm of the interatrial septum. For the purposes of this review, we analysed our experience using the Amplatzer device in 150 patients with interatrial communications. Of these, 104 had a defect within the oval fossa, 33 a patent oval foramen, and 13 had undergone fenestration of a Fontan procedure. Of those with defects within the oval fossa, a device was implanted in 100 patients, and 2 of these patients subsequently required surgical intervention, 1 because of migration and the other because of malformation of the device. Of the remaining 98 patients, complete occlusion has been achieved in 90% at 1 year. Any residual leaks are either trivial or small. In those with a patent oval foramen, the septal occluder was used to close 20, whilst the device designed specifically for this purpose was used in 13. On follow-up contrast echocardiography, only 2 patients have a small residual right-to-left shunt. Complete occlusion was achieved for all the Fontan fenestrations, although 1 patient later underwent surgery for baffle dehiscence. Other significant complications occurred in 2 patients who developed deep vein thrombosis, and 3 patients who suffered transient supraventicular arrhythmias. Although the Amplatzer device has been in clinical use for only 3 years, its unique design, and ease of use, has resulted in its widespread adoption by many centres. The results to date are very encouraging, but it must be remembered that there is, as yet, no long-term follow up data available for this life-long implant. PMID- 11049126 TI - Evaluation of the morphology of the oval fossa for placement of devices. AB - OBJECTIVES: First, to examine the morphology of heart specimens with defects of the oval fossa so as to define the factors that facilitate appropriate selection of the size of devices used for inteventional closure. Second, to examine the relationship between morphology and transthoracic and transesophageal echocardiography. BACKGROUND: The success of transcatheter closure is influenced by the variable morphology of deficiencies with the oval fossa, and of the relationship of the fossa itself to adjacent structures. More appropriate selection could reduce the incidence of failures. METHODS: From over 100 specimens in the cardiac registry at the University of California, San Francisco, we judged 16 hearts with atrial septal defects within the oval fossa, either in isolation or associated with other cardiac malformation, to be suitable for this study. We measured the dimensions of the defect and the surrounding rims of the fossa. All values were normalized to the diameter of the aortic root. RESULTS: A fenestrated defect was present in 9 specimens (56%). The shape defect itself was oval in all specimens, with a ratio of major to minor axes of 1.70 + 0.63. The major axis took one of three main directions with respect to the vertical plane: in 11 specimens (69%o) it was at horizontal; in 3 (19%) it was at oblique at an angle of 45 degrees; and in 2 (12%) it was vertical. Discordance was noted in some hearts between the major axis of the defect and that of the oval fossa. Structures closest to the rim of the fossa were the aortic mound, the coronary sinus, and the hinge point of the aortic leaflet of the mitral valve. CONCLUSIONS: Extrapolating from these specimens permitted identification of the major and minor axes of the atrial septal defect by transthoracic and transesophageal echocardiography. Our study has identified landmarks and dimensions that may be employed to improve effectiveness of selection of patients for transcatheter closure of defects within the oval fossa. PMID- 11049127 TI - Transesophageal echocardiographic predictors for successful transcatheter closure of defects within the oval fossa using the CardioSEAL septal occlusion device. AB - OBJECTIVES: To define the utility of transesophageal echocardiography in predicting the likelihood of a successful procedure and residual shunting in patients undergoing transcatheter closure of defects within the oval fossa using the CardioSEAL device. BACKGROUND: Transesophageal echocardiography is used to monitor transcatheter closure of interatrial defects within the the oval fossa, but predictors of successful closure and residual shunting have yet to be determined. METHODS: We reviewed transesophageal echocardiograms obtained from 26 consecutive patients undergoing attempted transcatheter closure of interatrial defects within the oval fossa between January, 1997 and May, 1998. Assessment of the atrial septum, the septal defect, and the rims of the oval fossa bordering the defect was performed in 3 planes: longitudinal, 4-chamber, and basal short axis. RESULTS: Closure proved successful in 24 patients (92%). The defect was significantly larger, and the anterosuperior rim of the defect smaller, in the 2 patients in whom occlusion was not successful. Residual shunting 24 hrs after closure was detected in 14 patients. Significant predictors of leakage included smaller posterior and superior rims, a larger shunt prior to closure, and herniation of a one left atrial arm of the device into the right atrium. In all cases, the sites of leakage were the superior rim of the defect at the superior cavo-atrial junction, and the anterosuperior rim behind the aortic root. Herniation of a left atrial arm into the right atrium was seen in 7 patients (29%). In all, it was the anterosuperior arm which herniated Doppler color flow was suboptimal in detecting residual leaks, and was enhanced substantially with the use of contrast echocardiography. CONCLUSIONS: Transesophageal echocardiography allows excellent assessment of the oval fossa and deficiencies of its floor in all of their dimensions. It is an important tool for guiding the deployment of the occlusion device in patients undergoing attempted transcatheter closure of defects within the fossa. Contrast echocardiography should be used for optimal detection of residual shunting. PMID- 11049128 TI - A European multicentric experience using the CardioSEal and Starflex double umbrella devices to close interatrial communications holes within the oval fossa. AB - In this review, we describe the experience from 13 European centres using the CardioSEAL and Starflex double umbrella devices to close interatrial communications within the oval fossa (so-called 'secundum' defects). Between October 1996 and April 1999, the procedure was attempted in 334 patients with a mean age of 12 years and a mean weight of 44kg. The mean measured stretched diameter of the defect was 15 mm. In the overall group, the defect was solitary in 245 patients (73%), multiple in 21 (6%), associated with an aneurysm of the flap valve in 15 (5 %), was represented by patency of the oval foramen in 44 (13%), and was a fenestration in a Fontan repair in 9 (3%). In all patients, the devices were inserted under general anesthesia, using fluoroscopic and transesophageal echocardiographic control. Implantation was achieved in 325 (97,3%). The device embolized within either a few minutes or a few hours in 13 patients (4%). Of these, uncomplicated surgical repair was undertaken in 10, while the device was retrieved in 3 using catheters and a second device was successfully implanted. Residual shunting was detected immediately after the procedure in 41% of the patients, with the incidence decreasing to 31% at discharge, 24% at 1 month, 21% at 6 months, and 20.5% at one year. During the period of follow-up, elective surgical repair became necessary in two patients, due to malposition of the device in one, and late embolization in the other. Fractures of arms were seen in 6.1 %, most commonly with the largest devices. All those with fractured arms of the device were asymptomatic, and no clinical complications related to the fractures were observed. There were no arrhythmias, endocarditis, valvar distortion, thromboembolic events, or other complications. After one year of follow-up, clinical success, defined as complete closure of the defect or presence of only a trivial leak, had been obtained in 92.5% of the patients. We conclude, therefore, that these devices produce excellent results when used to close defects of small to moderate size. Results are less than optimal, or else complications ensure, when attempts are made to close very large defects. PMID- 11049129 TI - Experience in one centre using the buttoned device for occlusion of atrial septal defect: comparison with the Amplatzer septal occluder. AB - We report our experience using the buttoned device to close defects within the oval fossa and probe-patent oval foramens, comparing the findings with those obtained with the Amplatzer septal occluder. From 1992 to 1997, we used the buttoned device to close defects in 73 consecutive patients, 64 with defects in the oval fossa and nine with patent foramens. We compared this experience with a further series of 62 patients seen from 1997 to 1999 in whom the Amplatzer septal occluder was used. Successful implantation was achieved in three-quarters of those with septal defects in whom the buttoned device was used, in all of those in whom the buttoned device was used for patent foramens, and in nine-tenths of those in whom closure was attempted using the Amplatzer occluder. Immediate surgery was needed in 3 patients in whom a buttoned device was used, one because of embolization and two with residual shunts and a straddling device. Similar immediate surgery was needed to retrieve one embolized Amplatzer occluder. During follow-up, surgery was needed in a further 7 patients, all having had insertion of a buttoned device, because of atrial perforation in one and a significant residual shunt in the remainder. At late follow-up, the rate of complete occlusion was 69% in the patients in whom the buttoned device was used to close a septal defect, 100% when the buttoned device was used for patent foramens, and 95% in those treated with the Amplatzer occluder. Our experience shows that the Amplatzer occluder produced a significantly higher rate of occlusion for larger defects, and with a shorter fluoroscopy time than the buttoned device. The Amplatzer septal occluder, therefore, is our preferred device for closure of defects within the oval fossa. PMID- 11049130 TI - Treatment of atrial septal defects in symptomatic children aged less than 2 years of age using the Amplatzer septal occluder. AB - AIMS: To assess results of closure of atrial septal defects within the oval fossa by devices delivered by catheterisation in symptomatic infants and children under 2 years of age. METHODS AND RESULTS: The Amplatzer septal occluder was used. Results, and complications of transcatheter device treatment in patients aged below 2 years were compared to previous results from our institution. PATIENTS: We attempted closure in 12 consecutive patients below the age of 2 years who presented with an atrial septal defect between May 1997 and 1999. Symptoms were failure to thrive in 6, frequent chest infections in 5, and the need for treatment of heart failure in the other. All were thought to have a defect suitable for interventional closure. The atrial defects were seen in isolation in 10 children, but 2 had associated pulmonary stenosis which had been treated by balloon dilation prior to placement of the Amplatzer occluder. RESULTS: The Amplatzer septal occluder was implanted at a mean age of 1.4 +/- 0.4, with a range from 0.8 to 1.8 years. Ratios of pulmonary-to-systemic flow had been 2.1 +/ 0.5, with a range from 1.6 and 3.2, and the defect was measured at 12 +/- 4 mms. Fluoroscopy time was 12.8 +/- 10.2 minutes, with a range from 5 to 43 minutes, and the time of the overall procedure was 162 +/- 70 minutes, with a range from 85 to 360 minutes. It proved necessary to remove the device in 2 patients (16%) because of a residual shunt and movement after release. One of these developed transient neurological complications. Both subsequently underwent surgical treatment. CONCLUSION: Symptomatic patients less than 2 years of age can undergo successful closure of an atrial septal defect using the Amplatzer device, but the rates of success are less, and procedure time longer, than in older children or adults. PMID- 11049131 TI - An aortic valve with four leaflets. PMID- 11049132 TI - Cardiac tamponade in a child with ascariasis. AB - Ascaris infection, when severe, may manifest with constitutional symptoms. The parasites penetrate the intestinal walls, reach the venules and lymphatics and, through the portal circulation, may affect the heart. To the best of our knowledge, involvement of the pericardium has yet to be reported. We describe here a case of severe pericardial effusion and cardiac tamponade in a child with ascariasis. PMID- 11049133 TI - Carrying out inteventional procedures, and when the arterial duct is patent. PMID- 11049134 TI - Warm up. PMID- 11049135 TI - Seduction. PMID- 11049136 TI - Factors contributing to low back pain in rowers. PMID- 11049137 TI - Why exercise in paraplegia? PMID- 11049138 TI - Magnetic resonance technology in training and sports. PMID- 11049139 TI - Stretching before exercise: an evidence based approach. PMID- 11049140 TI - Is exercise effective treatment for osteoarthritis of the knee? AB - OBJECTIVE: To review and determine the effectiveness of exercise treatment in osteoarthritis of the knee. METHODS: A computerised literature search of Medline was carried out searching between June 1966 and January 2000. RESULTS: Twenty three randomised controlled trials were identified from the literature. Only three trials were sufficiently powered. Small to moderate beneficial effects of exercise treatment were found for pain, small beneficial effects on disability outcome measures, and moderate to great beneficial effects were observed according to patient global assessment of effect. It was not possible to obtain evidence on the content of exercise interventions, as studies were hampered by lack of attention to proper concealment, reporting of adverse effects, and long term effects of exercise treatment. The lack of standard outcomes measures is also noted. CONCLUSIONS: The available evidence indicates beneficial short term effects of exercise treatment in patients with osteoarthritis of the knee. However, the number of available studies is limited, and more research is needed to expand this recommendation. Specifically, additional trials should provide information on adherence, home based interventions, interaction with pharmacological treatments, functional outcomes measures relevant to exercise treatment in these patients, and long term effects. At present, doctors should recommend exercise to all patients with mild/moderate disease. Further study should be encouraged and exercise should be continued to be recommended as a mainstay of non-pharmacological treatment of osteoarthritis of the knee. PMID- 11049141 TI - Long term health impact of playing professional football in the United Kingdom. AB - OBJECTIVE: To describe the long term impact of football on the health related quality of life (HRQL) of former professional footballers in the United Kingdom. METHOD: A cross sectional survey gathered data from 284 former professional players. Respondents reported medical treatments, osteoarthritis (OA) diagnosis, other morbidity, disability status, and work related disability since their football career. The EuroQol (EQ-5D) and global health rating scales were selected to assess HRQL. RESULTS: Medical treatment for football related injuries was a common feature, as was OA, with the knee being the most commonly affected joint. Respondents with OA reported poorer HRQL compared with those without OA. As with medical treatments and problems on each of the five EQ-5D dimensions (pain, mobility, usual activities, anxiety/depression, self care), frequency of disability and work related disability were higher among respondents with OA than those without. CONCLUSION: This exploratory study suggests that playing professional football can impact on the health of United Kingdom footballers in later life. The development of OA was associated with poorer outcomes on all aspects of HRQL. PMID- 11049142 TI - Impact energy attenuation performance of football headgear. AB - OBJECTIVES: Commercially available football head protectors were tested to determine their impact energy attenuation performance and ability to reduce the likelihood of concussion. METHODS: Prospective study using standardised impact test methods with both rigid (magnesium) and Hybrid III headforms. RESULTS: Eight commercially available head protectors from six manufacturers were tested. The magnitude of the headform accelerations increased as the drop height was increased, ranging from a minimum of 64 g from a height of 0.2 m to a maximum of 1,132 g from a height of 0.6 m. The head injury criterion and maximum headform acceleration values followed a similar trend. A steep increase was noted in the magnitude of maximum headform acceleration and head injury criterion when the drop height was increased from 0.4 to 0.5 m. This indicates that the foam material was completely compressed at an impact energy above about 20 J and therefore offers little protection against impacts of greater severity. Repeated tests using a drop height of 0.3 m showed that some helmets exhibit a "memory" effect, whereby impact performance is reduced by up to 50% with repeated impacts. CONCLUSIONS: Laboratory tests indicate that current commercially available football headgear performance will not reduce the likelihood of concussion. The absence of internationally recognised standards for soft headgear designed to ameliorate concussion is a major deficiency in sports injury prevention. PMID- 11049143 TI - Validation of an instrument for injury data collection in rugby union. AB - OBJECTIVE: To provide the basis for collecting rugby union injury data using a rigorously validated injury report form. METHODS: Seven stages were used to assess face, content, and criterion validity of the rugby union injury report form. A 22 member panel plus four sporting bodies assessed the form for face validity, and an expert panel assessed it for content and criterion validity. Panel members were consulted until consensus was reached. A yardstick developed by an expert panel using the Delphi technique was used to assess the reliability of the form. An independent panel of 10 viewed a series of five videotaped injuries, three times over a five week period to assess inter-rater and intrarater reliability. The form was then trialed by 40 people in situ during four games. RESULTS: The rugby union injury report form for games and training was developed, and the face, content, and criterion validity successfully assessed. A seven step protocol to create a yardstick was also developed to assist in the validation process. Both inter-rater and intrarater reliability results indicated a 98% agreement. The 40 trialists who completed forms in situ during four games were found to have an inter-rater reliability agreement of 98% for nine injuries. CONCLUSIONS: A measurement instrument for injury data collection in rugby union was successfully developed and validated, providing researchers with a basis for future studies in this area. A procedure to develop future injury data collection instruments in other sports was also developed. PMID- 11049144 TI - Impact of professionalism on injuries in rugby union. AB - OBJECTIVES: To measure the frequency and nature of injuries occurring in competitive matches since professionalism was introduced in rugby union. METHODS: The cohort study previously conducted in players from senior rugby clubs in the Scottish Borders in 1993-1994 when rugby union was an entirely amateur sport was repeated in 1997-1998. The same injury definition, outcome criteria, and method of calculating playing hours were used. In total, 803 (84%) of 960 eligible players participated, including all 30 adult players who played professionally for the Scottish Rugby Union or Border Reivers District. The 576 injury episodes in 381 of these players in competitive matches were compared with the 373 injuries in 266 players out of 975 (94%) who were eligible and registered with the same senior rugby clubs in 1993-1994. Outcomes were the occurrence of injury episodes, days away from playing or training for rugby, and time lost to employment or attendance at school/college as a consequence of being injured. RESULTS: The proportion of players who were injured almost doubled from 1993-1994 to 1997-1998, despite an overall reduction of 7% of the playing strength of participating clubs. Period prevalence injury rates rose in all age specific groups, particularly in younger players. This translated into an injury episode every 3.4 matches in 1993-94, rising to one in every 2.0 matches in 1997-1998. An injury episode occurred in a professional team for every 59 minutes of competitive play. Professional players sustained a higher proportion of recurrent injuries, particularly in the early part of the season. Some 56% of all their days lost to the game were caused by injuries to the muscles, ligaments, and joints of the knee, hip, and thigh. CONCLUSIONS: The introduction of professionalism in rugby union has coincided with an increase in injuries to both professional and amateur players. To reduce this, attention should be focused on the tackle, where many injuries occur. The International Rugby Board should place a moratorium on the use of protective equipment in competitive matches until its contribution to player morbidity has been fully assessed. PMID- 11049145 TI - Functional instability in non-contact ankle ligament injuries. AB - OBJECTIVES: To measure objectively functional standing balance in the acute stages of non-contact ankle sprain, and to compare patients with controls. METHODS: A Chattanooga balance machine was used to measure postural stability in patients with acute ankle sprain and uninjured controls over a two week period, in one and two legged stance, with eyes open and closed. Participants also completed the Olerud and Molander questionnaire to provide a subjective measure of ankle function. RESULTS: There was a highly significant improvement in questionnaire scores for the patients during the study period (p<0.0001). Patients appeared to be less stable than controls in all balance tests, although the difference did not reach significance. There was evidence of improvement over time in the number of tests successfully completed on the injured leg in single legged stance with eyes closed (p = 0.043) between visits 1 and 3. CONCLUSIONS: The patient group showed a subjective improvement, which supports clinical experience of treating acute ankle injuries. There is some evidence that on average the patient group appeared to be less stable than controls in all balance tests, although the difference did not reach statistical significance, even on the uninjured leg. There is a need to carry out further studies to confirm the results found in this pilot study and to investigate the hypotheses generated. It would be useful to evaluate a simple test that could be used clinically to monitor progress after ankle injury, and also to identify athletes with decreased functional stability, who may be more at risk of sustaining ankle injury. PMID- 11049146 TI - Physiological and anthropometric determinants of sport climbing performance. AB - OBJECTIVE: To identify the physiological and anthropometric determinants of sport climbing performance. METHODS: Forty four climbers (24 men, 20 women) of various skill levels (self reported rating 5.6-5.13c on the Yosemite decimal scale) and years of experience (0.10-44 years) served as subjects. They climbed two routes on separate days to assess climbing performance. The routes (11 and 30 m in distance) were set on two artificial climbing walls and were designed to become progressively more difficult from start to finish. Performance was scored according to the system used in sport climbing competitions where each successive handhold increases by one in point value. Results from each route were combined for a total climbing performance score. Measured variables for each subject included anthropometric (height, weight, leg length, arm span, % body fat), demographic (self reported climbing rating, years of climbing experience, weekly hours of training), and physiological (knee and shoulder extension, knee flexion, grip, and finger pincer strength, bent arm hang, grip endurance, hip and shoulder flexibility, and upper and lower body anaerobic power). These variables were combined into components using a principal components analysis procedure. These components were then used in a simultaneous multiple regression procedure to determine which components best explain the variance in sport rock climbing performance. RESULTS: The principal components analysis procedure extracted three components. These were labelled training, anthropometric, and flexibility on the basis of the measured variables that were the most influential in forming each component. The results of the multiple regression procedure indicated that the training component uniquely explained 58.9% of the total variance in climbing performance. The anthropometric and flexibility components explained 0.3% and 1.8% of the total variance in climbing performance respectively. CONCLUSIONS: The variance in climbing performance can be explained by a component consisting of trainable variables. More importantly, the findings do not support the belief that a climber must necessarily possess specific anthropometric characteristics to excel in sport rock climbing. PMID- 11049147 TI - The slow component of VO2 in professional cyclists. AB - OBJECTIVES: To analyse the slow component of oxygen uptake (VO2) in professional cyclists and to determine whether this phenomenon is due to altered neuromuscular activity, as assessed by surface electromyography (EMG). METHODS: The following variables were measured during 20 minute cycle ergometer tests performed at about 80% of VO2MAX in nine professional road cyclists (mean (SD) age 26 (2) years; VO2max 72.6 (2.2) ml/kg/min): heart rate (HR), gas exchange variables (VO2, ventilation (VE), tidal volume (VT), breathing frequency (fb), ventilatory equivalents for oxygen and carbon dioxide (VE/VO2 and VE/VCO2 respectively), respiratory exchange ratio (RER), and end tidal PO2 and PCO2 (PETO2 and PETCO2 respectively)), blood variables (lactate, pH, and [HCO3-]) and EMG data (root mean from square voltage (rms-EMG) and mean power frequency (MPF)) from the vastus lateralis muscle. RESULTS: The mean magnitude of the slow component (from the end of the third minute to the end of exercise) was 130 (0.04) ml in 17 minutes or 7.6 ml/min. Significant increases from three minute to end of exercise values were found for the following variables: VO2 (p<0.01), HR (p<0.01), VE (p<0.05), fb (p<0.01), VE/VO2 (p<0.05), VE/VCO2 (p<0.01), PETO2 (p<0.05), and blood lactate (p<0.05). In contrast, rms-EMG and MPF showed no change (p>0.05) throughout the exercise tests. CONCLUSIONS: A significant but small VO2 slow component was shown in professional cyclists during constant load heavy exercise. The results suggest that the primary origin of the slow component is not neuromuscular factors in these subjects, at least for exercise intensities up to 80% of VO2MAX. PMID- 11049149 TI - The sliding stop: a technique of fielding in cricket with a potential for serious knee injury. AB - The sliding stop method of fielding in cricket is gaining popularity in schools and club cricket through its frequent exposure on television. The case history is reported of a cricketer who suffered a torn medial meniscus in his knee, a rare cricketing injury, while performing this technique incorrectly in a club game. The correct method of performing the technique is described in coaching manuals but is not commonly instructed at club or school level. The sliding stop should be discouraged in school and for club cricketers unless appropriately coached. PMID- 11049148 TI - Medical supervision of sport diving in Scotland: reassessing the need for routine medical examinations. AB - BACKGROUND: The value of diving medicals in preventing incidents is uncertain and there has been only limited evaluation of the fitness to dive guidelines in a sport diving population. OBJECTIVE: To examine the need for routine diving medical examinations in the Scottish Sub-Aqua Club (SSAC) between 1991 and 1998. METHODS: A medical examination of all SSAC divers is performed at entry and then every one to five years based on their age and medical condition This information was analysed in terms of questionnaire findings and examination abnormalities. RESULTS: There were 2,962 medical forms available for analysis. Examination abnormalities were found in 174 subjects (5.9% of the population), with obesity affecting 75 subjects (2.5%). There was a linear increase in mean body mass index (r2 = 0.92), and a significant difference between 1991 and 1998 (mean (SD) of 24.1 (3.07) and 25.02 (3.4) respectively, p = 0.002) which was not related to age or sex distribution. There was also a significant increase in the prevalence of smokers (chi2 = 4.02, p = 0.045). The most common specialist referral was for evaluation of asthma, with hypertension and obesity as the next most common reasons. Most subjects were allowed to dive, with only 43 (25%) being failed outright. Overall, no examination abnormality alone caused a subject to be classified unfit to dive, and referrals were prompted by the answers in the questionnaire. CONCLUSIONS: No significant unexpected abnormalities were found on clinical examination in this population of sport divers. Conditions that prevented subjects from diving were detected by the questionnaire, and this prompted referral for further assessment by the medical advisors. Routine medical examinations were of little value. PMID- 11049150 TI - Frozen chips: an unusual cause of severe frostbite injury. AB - A case of severe frostbite injury to the right foot is presented. This was caused by the inappropriate application of a bag of frozen chips to the foot in an attempt to ease non-specific pain. No specific acute traumatic injury was identified. As the patient was a teacher of physical education, the pain had initially been assumed to originate from a minor musculoskeletal injury. Full recovery ensued after surgical excision of necrotic tissue and split skin grafting. The danger of inappropriate overenthusiastic use of ice packs or other frozen material to treat soft tissue injuries is emphasised. The need for education to prevent similar future injuries is discussed. PMID- 11049151 TI - Physiological and biomechanical adaptations to the cycle to run transition in Olympic triathlon: review and practical recommendations for training. AB - Current knowledge of the physiological, biomechanical, and sensory effects of the cycle to run transition in the Olympic triathlon (1.5 km, 10 km, 40 km) is reviewed and implications for the training of junior and elite triathletes are discussed. Triathlon running elicits hyperventilation, increased heart rate, decreased pulmonary compliance, and exercise induced hypoxaemia. This may be due to exercise intensity, ventilatory muscle fatigue, dehydration, muscle fibre damage, a shift in metabolism towards fat oxidation, and depleted glycogen stores after a 40 km cycle. The energy cost (CR) of running during the cycle to run transition is also increased over that of control running. The increase in CR varies from 1.6% to 11.6% and is a reflection of triathlete ability level. This increase may be partly related to kinematic alterations, but research suggests that most biomechanical parameters are unchanged. A more forward leaning trunk inclination is the most significant observation reported. Running pattern, and thus running economy, could also be influenced by sensorimotor perturbations related to the change in posture. Technical skill in the transition area is obviously very important. The conditions under which the preceding cycling section is performed-that is, steady state or stochastic power output, drafting or non-drafting-are likely to influence the speed of adjustment to transition. The extent to which a decrease in the average 10 km running speed occurs during competition must be investigated further. It is clear that the higher the athlete is placed in the field at the end of the bike section, the greater the importance to their finishing position of both a quick transition area time and optimal adjustment to the physiological demands of the cycle to run transition. The need for, and current methods of, training to prepare junior and elite triathletes for a better transition are critically reviewed in light of the effects of sequential cycle to run exercise. PMID- 11049153 TI - Injuries of the sporting knee PMID- 11049152 TI - East African running dominance: what is behind it? PMID- 11049155 TI - Pharmacological treatment of significant cardiac arrhythmias. PMID- 11049154 TI - Knee instability: isolated and complex. PMID- 11049156 TI - Bone density in elderly women. PMID- 11049157 TI - Who should be blinded? PMID- 11049158 TI - The genetics of physical fitness. PMID- 11049159 TI - Serum concentrations of P-selectin decline rapidly in resting humans. PMID- 11049160 TI - Exercise at altitude. PMID- 11049161 TI - "The branches into which bacteriology is now ramifying" revisited. AB - The American Society for Microbiology was originally founded in 1899 as the Society of American Bacteriologists. The transition from "bacteriology" to "microbiology" and from an emphasis on the identity of the membership (bacteriologists) to an emphasis on the discipline (microbiology) was a contentious one that occurred in several steps. This article reviews the history and events that accompanied this development. PMID- 11049162 TI - Welch, Sedgwick, and the Hopkins model of hygiene. AB - William H. Welch and William T. Sedgwick, two of the founding fathers of American public health, were both early generation "Hopkins Men." Sedgwick was part of the first group of graduate students to attend Johns Hopkins University, and Welch was part of the initial faculty at the University's medical school. While they never worked together as colleagues at Hopkins, both became interested in the exciting new discoveries of the microbial nature of human disease and developed similar public health programs based on this information. Sedgwick expanded upon these investigations in the new sanitary science program at MIT, where academic public health first emerged in the United States following Sedgwick's appointment in 1883. Welch, who had been exposed to European research in microbiology, promoted microbial research in pathology in Baltimore in 1884. His laboratory based investigations expanded until they led to the formation of the country's first school of public health in 1916. Thus, a "Hopkins Model" for hygiene and public health emerged from the efforts of both Welch and Sedgwick. PMID- 11049163 TI - Evolution of microbiology as seen in the textbooks of Edwin O. Jordan and William H. Park. AB - Historians of science account the appearance of textbooks as an important step in the formation and consolidation of a new discipline. The texts of Park and Jordan were both very important in this light; however; they also can be used as a gauge of changing concepts within microbiology in the first four decades after its consolidation as a discipline, 1900-1940. This paper tracks these important texts and through them changing attitudes toward several important concepts: bacterial variation, human/bovine tuberculosis, and the existence of a non-symptomatic carrier state in infectious disease. The two texts are also compared regarding their view of microbes as pathogens vs. microbes as important and ubiquitous ecological agents. PMID- 11049164 TI - Robert Earle Buchanan: an unappreciated scientist. AB - Robert Earle Buchanan (1883-1973), 19th President of the Society of American Bacteriologists (later American Society for Microbiology), was one of the more important 20th century microbiologists. He was a dominant force in creating the field of bacterial systematics and made significant contributions to microbial physiology. He also numbered a number of influential textbooks. A reasonable conclusion is that Buchanan was a major cultivator of modern microbiology. To justify that assertion, I have four major objectives in this essay: i) a brief biographical review of Buchanan's early life; ii) a brief review of his scientific contributions, many of which go beyond his recognized contributions to bacterial systematics; iii) Buchanan was an important academic administrator who created the microbiology program and fostered a strong graduate education program at Iowa State, iv)finally, I close the essay with a focus on Buchanan's "moral character." PMID- 11049165 TI - Hans Zinsser: a tale of two cultures. AB - Hans Zinsser, president of the Society of American Bacteriologists in 1926, was known as much for his literary and textbook writing as for his scientific contributions. He was a widely known scientist and person of letters. His early interests in poetry and other forms of literature were maintained and developed during his career as a microbiologist, and his most enduring legacy is based on his writing about microbiology for a general readership as well as his reflective and philosophical autobiography. PMID- 11049166 TI - Alice C. Evans: breaking barriers. AB - Despite severe and persistent criticism of her research, Alice Evans persevered in her pioneering work on the bacterial contamination of milk, identifying the organism that caused undulant fever and demonstrating that drinking unpasteurized cow's milk could transmit the disease, undulant fever, to humans. The opprobrium that Alice Evans endured was unrelenting, even after her election as the first President of the Society of American Bacteriologists, (now the American Society for Microbiology), but she remained undeterred, a true heroine of American microbiology and a magnificent public health worker. PMID- 11049167 TI - One hundred years of presidents of SAB/ASM: the Rockefeller connection. PMID- 11049168 TI - On beam quality and stopping power ratios for high-energy x-rays. AB - The aim of this work is to quantitatively compare two commonly used beam quality indices, IPR(20/10) and %dd(10)x, with respect to their ability to predict stopping power ratios (water to air), s(w,air), for high-energy x-rays. In particular, effects due to a varied amount of filtration of the photon beam will be studied. A new method for characterizing beam quality is also presented, where the information we strive to obtain is the moments of the spectral distribution. We will show how the moments enter into a general description of the transmission curve and that it is possible to correlate the moments to s(w,air) with a unique and simple relationship. Comparisons with TPR(20/10) and %dd(10), show that the moments are well suited for beam quality specification in terms of choosing the correct s(w,air). PMID- 11049169 TI - Photon scatter in intensity modulating filters evaluated by first Compton scatter and Monte Carlo calculations and experiments in broad beams. AB - High-atomic-number materials may be used as intensity modulating filters for inverse radiation treatment planning with photon beams. Such filters, when placed in a bremsstrahlung beam, attenuate the primary fluence, but also produce scattered photons that will reach the patient. To account for such effects in the optimization of photon beam intensities a semiempirical method based on narrow and broad beam transmission measurements was used to quantify the number of scattered photons produced in these filters. The method was verified by performing analytical calculations based on first scatter and a Monte Carlo simulation in 6 and 18 MV photon beams. The resultant experimental transmission ratios agree with calculations by these methods within 2 per cent under the experimental conditions investigated. The semiempirical method can thus be used as a basis for preliminary decision-making to select the proper material for intensity modulating filters and can provide a fast method to perform independent quality checks of the calculation accuracy of dose planning systems. Change in beam penetration is of less concern when treatments of target volumes at smaller depths are of interest. A 10 g cm(-2) thick filter made of low-melting-point alloy produces a change in percentage depth dose of less than 2 per cent for depths larger than 10 cm independent of field size. Similarly the scatter correction modifies the dose distribution by less than 5-10 per cent in most cases. PMID- 11049170 TI - Dose distribution measurements by MRI of a phantom containing lung tissue equivalent compartments made of ferrous sulphate gel. AB - Gel dosimetry based on magnetic resonance imaging (MRI) has previously been shown to provide verification of calculated dose distributions in soft tissue equivalent homogeneous phantoms. This study demonstrates how measurements of dose distribution can also be achieved in a phantom containing porous, lung equivalent, Fricke gel. A phantom was made of Fe2+ infused low-density gel and conventional ferrous sulphate gel, filled in separate compartments in a Perspex container. Absorbed dose measurements were accomplished by MR imaging and by calibrating the proton spin-lattice relaxation rate (R1) versus absorbed dose by means of TLD measurements. This study shows that the production of lung equivalent low-density (LD) dosimeter gel (mean CT number of -610 HU) is feasible. The MR signal detected in the LD gel dosimeter was substantially more noisy (i.e. displayed larger random fluctuations) than the signal from conventional gel, as expected. A deviation between calculated (TPS) and measured dose of about 3% (6 MV) and 4-7% (15 MV) was found in the LD region of the phantom. These results correspond well with data from other studies of dose distribution in lung-equivalent phantoms. The Fe2+ infused LD gel therefore seems suitable for measurement of absorbed dose distribution in phantoms that contain lung tissue compartments. PMID- 11049171 TI - Improvement of internal dose calculations using mathematical models of different adult heights. AB - In internal dosimetry for both nuclear medicine and radiation protection, the adult morphology is represented by a limited number of anthropomorphic models that may not be suitable for all patients. To develop more patient-specific dosimetry, we derived six mathematical models for adults of different height. Three male models (160 cm, 170 cm and 180 cm) and three female models (150 cm, 160 cm and 170 cm), based on the MIRD model design, were developed from the statistical analysis of anthropometric data gathered from autopsies. Monte Carlo calculations were used to provide an example of estimations of S value for these new models for iodine 131 uniformly distributed successively in the stomach or in the urinary bladder. On average, for both male and female models, an increase in the model height of 10 cm leads to a mean reduction in the S value for iodine-131 by 20% and 29% when the stomach and the urinary bladder respectively are selected as source regions. Similarly, when the model height increases by 20 cm, the S values decrease on average by 35% and 48%. This study presents the use of anthropometric data to develop new mathematical models for adults of different height, and shows the significant influence of the morphology on dosimetric parameters. PMID- 11049172 TI - CT imaging based digitally reconstructed radiographs and their application in brachytherapy. AB - The aim of our study was to develop an algorithm to simulate the digitally reconstructed radiograph (DRR) calculation process for different beam qualities (photon energies) in the range 50 keV to 12 MeV. This was achieved using volumetric anatomical data for the patient obtained from three-dimensional diagnostic CT images. These DRR images can be used in three-dimensional treatment planning for external beam radiotherapy as well as for brachytherapy in the same way as conventional radiographic films. The advantages of using such DRRs in modern 3D brachytherapy treatment planning are shown. A number of tools are described, illustrating that the application of DRRs in brachytherapy is very useful. PMID- 11049173 TI - Non-iterative methods incorporating a priori source distribution and data information for suppression of image noise and artefacts in 3D SPECT. AB - Non-iterative methods have been developed for image reconstruction in 3D SPECT with uniform attenuation and distance-dependent spatial resolution. It was observed that these methods can, in general, be susceptible to data noise and other errors, yielding conspicuous image artefacts. In this work, we developed and evaluated a regularized inverse-filtering approach for effective suppression of noise and artefacts in 3D SPECT images without significantly compromising image resolution. The proposed approach allows the incorporation of a priori random image field and data information and can thus robustly control the degree of suppression of noise and artefacts in 3D SPECT images. Using computer simulations, we evaluated and compared quantitatively images reconstructed from data sets of various noise levels by the use of the proposed methods and the existing non-iterative methods. These numerical results clearly demonstrated that the proposed regularized inverse-filtering approach can effectively suppress image noise and artefacts that plague the existing non-iterative methods, thus yielding quantitatively more accurate 3D SPECT images. The proposed regularized inverse-filtering approach can also be generalized to other imaging modalities. PMID- 11049174 TI - A 2D/3D hybrid PET scanner with rotating partial slice-septa and its quantitative procedures. AB - This paper presents a PET scanner capable of acquiring projection data in three dimensional (3D) and two-dimensional (2D) modes simultaneously. The scanner has rotating partial slice-septa, and coincidence events are stored as 2D data or as 3D data depending on whether the lines of response are collimated by the septa or not. 68Ge/Ga rod sources can be set on the rotating septa, and a transmission scan for attenuation correction is performed in the 2D mode. The scanner allows simultaneous 3D-emission/2D-transmission scanning or post-injection transmission scanning with little cross-talk. A blank scan for detector normalization is also performed with the rotating rod sources in the 2D mode, from which we can derive the normalizing factors in both modes. The 3D/2D difference method is available for scatter correction, even in a dynamic study where the source distribution is changing. A 'summation method' is proposed as a new image reconstruction algorithm, in which the high- and low-frequency components of images are reconstructed from the 3D and 2D data respectively. In this method, most of the scatter contribution in the 3D data is removed by high-pass filtering, not by subtracting estimated scatter distribution, and hence the method is expected to be robust for scatter from outside the axial field of view. Computer simulations revealed that the rotating partial septa offer a single-scatter to true ratio similar to that of the conventional full septa if the depth of the partial septa is properly lengthened, with a small increase in multiple scattering. PMID- 11049175 TI - An automatic method to discriminate malignant masses from normal tissue in digital mammograms. AB - Specificity levels of automatic mass detection methods in mammography are generally rather low, because suspicious looking normal tissue is often hard to discriminate from real malignant masses. In this work a number of features were defined that are related to image characteristics that radiologists use to discriminate real lesions from normal tissue. An artificial neural network was used to map the computed features to a measure of suspiciousness for each region that was found suspicious by a mass detection method. Two data sets were used to test the method. The first set of 72 malignant cases (132 films) was a consecutive series taken from the Nijmegen screening programme, 208 normal films were added to improve the estimation of the specificity of the method. The second set was part of the new DDSM data set from the University of South Florida. A total of 193 cases (772 films) with 372 annotated malignancies was used. The measure of suspiciousness that was computed using the image characteristics was successful in discriminating tumours from false positive detections. Approximately 75% of all cancers were detected in at least one view at a specificity level of 0.1 false positive per image. PMID- 11049176 TI - Classification of reflectance spectra from pigmented skin lesions, a comparison of multivariate discriminant analysis and artificial neural networks. AB - Successful treatment of skin cancer, especially melanoma, depends on early detection, but diagnostic accuracy, even by experts, can be as low as 56% so there is an urgent need for a simple, accurate, non-invasive diagnostic tool. In this paper we have compared the performance of an artificial neural network (ANN) and multivariate discriminant analysis (MDA) for the classification of optical reflectance spectra (320 to 1100 nm) from malignant melanoma and benign naevi. The ANN was significantly better than MDA, especially when a larger data set was used, where the classification accuracy was 86.7% for ANN and 72.0% for MDA (p < 0.001). ANN was better at learning new cases than MDA for this particular classification task. This study has confirmed that the convenience of ANNs could lead to the medical community and patients benefiting from the improved diagnostic performance which can be achieved by objective measurement of pigmented skin lesions using spectrophotometry. PMID- 11049177 TI - Anisotropy of light propagation in human skin. AB - Using spatially resolved, steady state diffuse reflectometry, a directional dependence was found in the propagation of visible and near infrared light through human skin in vivo. The skin's reduced scattering coefficient mu(s)' varies by up to a factor of two between different directions of propagation at the same position. This anisotropy is believed to be caused by the preferential orientation of collagen fibres in the dermis, as described by Langer's skin tension lines. Monte Carlo simulations that examine the effect of partial collagen fibre orientation support this hypothesis. The observation has consequences for non-invasive diagnostic methods relying on skin optical properties, and it could be used non-invasively to determine the direction of lines of cleavage in order to minimize scars due to surgical incisions. PMID- 11049178 TI - A virtual linear accelerator for verification of treatment planning systems. AB - A virtual linear accelerator is implemented into a commercial pencil-beam-based treatment planning system (TPS) with the purpose of investigating the possibility of verifying the system using a Monte Carlo method. The characterization set for the TPS includes depth doses, profiles and output factors, which is generated by Monte Carlo simulations. The advantage of this method over conventional measurements is that variations in accelerator output are eliminated and more complicated geometries can be used to study the performance of a TPS. The difference between Monte Carlo simulated and TPS calculated profiles and depth doses in the characterization geometry is less than +/-2% except for the build up region. This is of the same order as previously reported results based on measurements. In an inhomogeneous, mediastinum-like case, the deviations between TPS and simulations are small in the unit-density regions. In low-density regions, the TPS overestimates the dose, and the overestimation increases with increasing energy from 3.5% for 6 MV to 9.5% for 18 MV. This result points out the widely known fact that the pencil beam concept does not handle changes in lateral electron transport, nor changes in scatter due to lateral inhomogeneitics. It is concluded that verification of a pencil-beam-based TPS with a Monte Carlo based virtual accelerator is possible, which facilitates the verification procedure. PMID- 11049179 TI - An orthogonal moment-based method for automatic verification of radiation field shape. AB - The purpose of this paper is to develop a new method for automated on-line verification of the treatment field shape during radiotherapy. The treatment field boundary is extracted from the digital portal image and is then approximated by a polygon. The proposed procedure used one of the approved field shapes as the reference boundary for automated comparison with subsequent portal field boundaries. The orthogonal moment-based method was applied to align treatment field boundaries that include the translational shifts, scaling factor and rotation angle. Firstly, the moments of order up to one were used to adjust the magnification and translation of the test field boundary related to the reference one; this step created a common coordinate system for the two images. Then a quadratic least-square objective function based on the orthogonal moments (e.g. Legendre moments) of the two field shapes was employed to perform rotational correction. Since moment computation by a straightforward method required a large number of multiplication and addition operations, a fast method for computing Legendre moments was also developed to decrease the calculation time. Application of the method to some simulated cases showed that our alignment procedure has an accuracy of 0.5 mm in detecting translational shift, 0.004 in detecting magnification and less than 0.3 degrees in detecting rotation angle between the test shape and the reference shape. The alignment procedure using the proposed method can be done within 2 s on a Pentium II personal computer. Therefore, our method is potentially useful for automated real-time treatment field shape verification. PMID- 11049180 TI - The energy-dependent electron loss model for pencil beam dose kernels. AB - The 'monoenergetic' electron loss model was derived in a previous work to account for pathlength straggling in the Fermi-Eyges pencil beam problem. In this paper, we extend this model to account for energy-loss straggling and secondary knock-on electron transport in order to adequately predict a depth dose curve. To model energy-loss straggling, we use a weighted superposition of a discrete number of monoenergetic pencil beams with different initial energies where electrons travel along the depth-energy characteristics in the continuous slowing down approximation (CSDA). The energy straggling spectrum at depth determines the weighting assigned to each monoenergetic pencil beam. Supplemented by a simple transport model for the secondary knock-on electrons, the 'energy-dependent' electron loss model predicts both lateral and depth dose distributions from the electron pencil beams in good agreement with Monte Carlo calculations and measurements. The calculation of dose distribution from a pencil beam takes 0.2 s on a Pentium III 500 MHz computer. Being computationally fast, the 'energy dependent' electron loss model can be used for the calculation of 3D energy deposition kernels in dose optimization schemes without using precalculated or measured data. PMID- 11049181 TI - Accuracy of the phase space evolution dose calculation model for clinical 25 MeV electron beams. AB - The phase space evolution (PSE) model is a dose calculation model for electron beams in radiation oncology developed with the aim of a higher accuracy than the commonly used pencil beam (PB) models and with shorter calculation times than needed for Monte Carlo (MC) calculations. In this paper the accuracy of the PSE model has been investigated for 25 MeV electron beams of a MM50 racetrack microtron (Scanditronix Medical AB, Sweden) and compared with the results of a PB model. Measurements have been performed for tests like non-standard SSD, irregularly shaped fields, oblique incidence and in phantoms with heterogeneities of air, bone and lung. MC calculations have been performed as well, to reveal possible errors in the measurements and/or possible inaccuracies in the interaction data used for the bone and lung substitute materials. Results show a good agreement between PSE calculated dose distributions and measurements. For all points the differences--in absolute dose--were generally well within 3% and 3 mm. However, the PSE model was found to be less accurate in large regions of low density material and errors of up to 6% were found for the lung phantom. Results of the PB model show larger deviations, with differences of up to 6% and 6 mm and of up to 10% for the lung phantom; at shortened SSDs the dose was overestimated by up to 6%. The agreement between MC calculations and measurement was good. For the bone and the lung phantom maximum deviations of 4% and 3% were found, caused by uncertainties about the actual interaction data. In conclusion, using the phase space evolution model, absolute 3D dose distributions of 25 MeV electron beams can be calculated with sufficient accuracy in most cases. The accuracy is significantly better than for a pencil beam model. In regions of lung tissue, a Monte Carlo model yields more accurate results than the current implementation of the PSE model. PMID- 11049182 TI - Monte Carlo characterization of clinical electron beams in transverse magnetic fields. AB - Monte Carlo simulations were employed to study the characteristics of the electron beams of a clinical linear accelerator in the presence of 1.5 and 3.0 T transverse magnetic fields and to assess the possibility of using magnetic fields in conjunction with modulated electron radiation therapy (MERT). The starting depth of the magnetic field was varied over several centimetres. It was found that peak doses of as much as 2.7 times the surface dose could be achieved with a 1.5 T magnetic field. The magnetic field was shown to reduce the 80% and 20% dose drop-off distance by 50% to 80%. The distance between the 80% dose levels of the pseudo-Bragg peak induced by the magnetic field was found to be extremely narrow, generally less than 1 cm. However, by modulating the energy and intensity of the electron fields while simultaneously moving the magnetic field, a homogeneous dose distribution with low surface dose and a sharp dose fall-off was generated. Heterogeneities are shown to change the effective range of the electron beams, but not eliminate the advantages of a sharp depth dose drop-off or high peak-to surface dose ratio. This suggests the applicability of MERT with magnetic fields in heterogeneous media. The results of this study demonstrate the ability to use magnetic fields in MERT to produce highly desirable dose distributions. PMID- 11049183 TI - Validation of a Monte Carlo dose calculation tool for radiotherapy treatment planning. AB - A new EGS4/PRESTA Monte Carlo user code, MCDOSE, has been developed as a routine dose calculation tool for radiotherapy treatment planning. It is suitable for both conventional and intensity modulated radiation therapy. Two important features of MCDOSE are the inclusion of beam modifiers in the patient simulation and the implementation of several variance reduction techniques. Before this tool can be used reliably for clinical dose calculation, it must be properly validated. The validation for beam modifiers has been performed by comparing the dose distributions calculated by MCDOSE and the well-benchmarked EGS4 user codes BEAM and DOSXYZ. Various beam modifiers were simulated. Good agreement in the dose distributions was observed. The differences in electron cutout factors between the results of MCDOSE and measurements were within 2%. The accuracy of MCDOSE with various variance reduction techniques was tested by comparing the dose distributions in different inhomogeneous phantoms with those calculated by DOSXYZ without variance reduction. The agreement was within 1.0%. Our results demonstrate that MCDOSE is accurate and efficient for routine dose calculation in radiotherapy treatment planning, with or without beam modifiers. PMID- 11049184 TI - Fast neutron absorbed dose distributions in the energy range 0.5-80 meV--a Monte Carlo study. AB - Neutron pencil-beam absorbed dose distributions in phantoms of bone, ICRU soft tissue, muscle, adipose and the tissue substitutes water, A-150 (plastic) and PMMA (acrylic) have been calculated using the Monte Carlo code FLUKA in the energy range 0.5 to 80 MeV. For neutrons of energies < or = 20 MeV, the results were compared to those obtained using the Monte Carlo code MCNP4B. Broad-beam depth doses and lateral dose distributions were derived. Broad-beam dose distributions in various materials were compared using two kinds of scaling factor: a depth-scaling factor and a dose-scaling factor. Build-up factors due to scattered neutrons and photons were derived and the appropriate choice of phantom material for determining dose distributions in soft tissue examined. Water was found to be a good substitute for soft tissue even at neutron energies as high as 80 MeV. The relative absorbed doses due to photons ranged from 2% to 15% for neutron energies 10-80 MeV depending on phantom material and depth. For neutron energies below 10 MeV the depth dose distributions derived with MCNP4B and FLUKA differed significantly, the difference being probably due to the use of multigroup transport of low energy (< 19.6 MeV) neutrons in FLUKA. Agreement improved with increasing neutron energies up to 20 MeV. At energies > 20 MeV, MCNP4B fails to describe dose build-up at the phantom interface and penumbra at the edge of the beam because it does not transport secondary charged particles. The penumbra width, defined as the distance between the 80% and 20% iso-dose levels at 5 cm depth and for a 10 x 10 cm2 field, was between 0.9 mm and 7.2 mm for neutron energies 10-80 MeV. PMID- 11049185 TI - Monte Carlo dose calculations and radiobiological modelling: analysis of the effect of the statistical noise of the dose distribution on the probability of tumour control. AB - The aim of this work is to investigate the influence of the statistical fluctuations of Monte Carlo (MC) dose distributions on the dose volume histograms (DVHs) and radiobiological models, in particular the Poisson model for tumour control probability (tcp). The MC matrix is characterized by a mean dose in each scoring voxel, d, and a statistical error on the mean dose, sigma(d); whilst the quantities d and sigma(d) depend on many statistical and physical parameters, here we consider only their dependence on the phantom voxel size and the number of histories from the radiation source. Dose distributions from high-energy photon beams have been analysed. It has been found that the DVH broadens when increasing the statistical noise of the dose distribution, and the tcp calculation systematically underestimates the real tumour control value, defined here as the value of tumour control when the statistical error of the dose distribution tends to zero. When increasing the number of energy deposition events, either by increasing the voxel dimensions or increasing the number of histories from the source, the DVH broadening decreases and tcp converges to the 'correct' value. It is shown that the underestimation of the tcp due to the noise in the dose distribution depends on the degree of heterogeneity of the radiobiological parameters over the population; in particular this error decreases with increasing the biological heterogeneity, whereas it becomes significant in the hypothesis of a radiosensitivity assay for single patients, or for subgroups of patients. It has been found, for example, that when the voxel dimension is changed from a cube with sides of 0.5 cm to a cube with sides of 0.25 cm (with a fixed number of histories of 10(8) from the source), the systematic error in the tcp calculation is about 75% in the homogeneous hypothesis, and it decreases to a minimum value of about 15% in a case of high radiobiological heterogeneity. The possibility of using the error on the tcp to decide how many histories to run for a given voxel size is also discussed. PMID- 11049186 TI - Proton stopping powers averaged over beam energy spectra. AB - Stopping powers averaged over the proton energy spectra at various depths in water and a tissue-like material were calculated for proton beams with initial energies between 50 and 250 MeV. The analysis was made for proton beams with a spread-out Bragg peak (SOBP) to assess in particular the variations of stopping power with depth in the SOBP plateau region. Nuclear interactions were accounted for in determination of the proton energy spectra. In modulated beams, stopping power varies considerably with depth along the plateau region of the spread-out peak. Moreover, stopping powers at the same water depth may differ from each other by up to 20% if referring to modulated or non-modulated beams. Calculations of water-air mass stopping power ratios were also performed, with and without the inclusion of nuclear interactions, for modulated and non-modulated beams. The stopping power ratios do not depend significantly on proton energy, and this makes the effect of accounting for nuclear interactions in proton fluence calculation negligible. For the same reason the stopping power ratios for modulated beams do not appreciably differ from those referred to non-modulated beams with the same Emax at the same depth in water. The present results also show that the dependence of stopping power on energy spread and spatial divergence for proton beams is not negligible in some conditions. Some conclusions are drawn on the possibility of using the residual range as a descriptor of the proton beam quality for different experimental beam conditions. PMID- 11049187 TI - Dosimetric characterization of silicon and diamond detectors in low-energy proton beams. AB - The dosimetric behaviour of a Scanditronix p-type silicon diode and a PTW natural diamond detector was studied in low-energy proton beams in the 8.3-21.5 MeV range. The properties investigated were linearity, reproducibility, dose rate dependence, energy and linear energy transfer (LET) dependence. The influence of detector thickness on the results of depth dose measurements was also demonstrated. A Markus parallel plate ionization chamber was used for reference dosimetry. Silicon diode and diamond detectors showed linearity at therapeutic dose level, reproducibility better than 1% (1sigma) and sensitivity variation with dose rate and proton energy. PMID- 11049188 TI - Fluence-to-dose conversion coefficients from monoenergetic neutrons below 20 MeV based on the VIP-man anatomical model. AB - A new set of fluence-to-absorbed dose and fluence-to-effective dose conversion coefficients have been calculated for neutrons below 20 MeV using a whole-body anatomical model, VIP-Man, developed from the high-resolution transverse colour photographic images of the National Library of Medicine's Visible Human Project. Organ dose calculations were performed using the Monte Carlo code MCNP for 20 monoenergetic neutron beams between 1 x 10(-9) MeV and 20 MeV under six different irradiation geometries: anterior-posterior, posterior-anterior, right lateral, left lateral, rotational and isotropic. The absorbed dose for 24 major organs and effective dose results based on the realistic VIP-Man are presented and compared with those based on the simplified MIRD-based phantoms reported in the literature. Effective doses from VIP-Man are not significantly different from earlier results for neutrons in the energy range studied. There are, however, remarkable deviations in organ doses due to the anatomical differences between the image-based and the earlier mathematical models. PMID- 11049189 TI - A novel method for real-time magnetic marker monitoring in the gastrointestinal tract. AB - In internal medicine, a simple method for the functional examination of the gastrointestinal tract without the risk of radiation exposure is required. We describe a novel principle based on the monitoring of magnetic markers which meets these demands. Our method employs a special permanent magnet which is repeatedly aligned by a vertically oriented pulsed magnetic field. Due to this alignment, the marker position can be derived from the stray field components measured by commercial field sensors. Our method was evaluated by means of a 3D intestinal phantom. The monitoring procedure yielded the time course of the marker position as a 3D plot either in real-time or as a time-lapse movie. The spatial resolution, expressed by the mean square deviation, was better than 10 mm and is thus sufficiently high to distinguish between adjacent loops of the gut. The temporal resolution, i.e. the minimum time between two successive measurements, was about 1 s. The presented method has very moderate technical demands and allows us to monitor magnetic markers in real-time. The technique may be useful with respect to functional examination of the gastrointestinal tract. In pharmaceutical research, our method offers the opportunity for remote drug release at any position of the gut. PMID- 11049190 TI - Conversion factors for the estimation of effective dose in paediatric cardiac angiography. AB - Conversion factors, which relate the kerma-area product to effective dose, have been estimated for paediatric cardiac x-ray angiography. Monte Carlo techniques have been used to calculate the conversion factors for a wide range of projection angles for children of five ages and for adults. Correction factors are provided so that the conversion factors can be adjusted for different tube potentials and filtrations. PMID- 11049191 TI - The interaction of MRI contrast agents with phospholipids. AB - The molecular interactions of three clinically used MRI contrast agents with lipid vesicles, consisting of egg phosphatidylcholine (EPC), have been studied using high-field NMR techniques. At a molar ratio of one contrast agent molecule to five phospholipid molecules, a significant increase in the proton resonance line width occurred for certain lipid head group moieties. A large decrease in the T1 relaxation times for the head group moieties was also observed. These two effects occurred regardless of the ionic status and the chelate structure of the three contrast agents. The structure of the contrast agents did, however, affect the magnitude of the two NMR parameter changes. These NMR effects also differed in magnitude amongst the various head group entities. The NMR effects were greatest for the head group moieties at or near the vesicle-water interface. The results are discussed in terms of the structure of the phospholipid-water interface. Since the use of contrast agents has become routine in clinical MRI, our results are of importance in terms of the interaction of the agents with physiological surfaces, many of which contain phospholipids. The understanding of such interactions should be of value not only for improved diagnostics, but also in the development of new contrast agents. PMID- 11049192 TI - Reconstruction of megavoltage photon spectra from electronic portal imager derived transmission measurements. AB - Three variants of the Schiff equation are investigated to model the spectra produced by megavoltage linear accelerators. These models are tested against well validated Monte Carlo (MC) generated spectra on the central axis of large-area fields, and show excellent agreement. Numerical reconstructions of 6 and 10 MV spectra using the same models are then presented, using experimental attenuation data derived from an electronic portal imager. The process of deriving spectra from experimental attenuation data is shown to be inherently badly constrained mathematically, with the derived spectrum being highly sensitive to noise in the source data, and non-unique. By placing a priori constraints on the Schiff model from both physical knowledge of the construction of the accelerator and MC data, physically useful results are gained and presented for both the energy dependence and off-axis behaviour of photon spectra. PMID- 11049193 TI - Effects of glass and backscatter on measurement of absorbed dose in polyacrylamide gel (PAG) dosimeters. AB - Different methodologies are used for calibrating polyacrylamide gel (PAG) dosimeters. One methodology involves injecting nitrogen-filled glass vials with polymer gel. Due to the vials being pre-filled with nitrogen, a nitrogen-filled space remains in the glass vial above the gel. The glass vial is then irradiated using ionizing radiation to polymerize the PAG. Monte Carlo simulations were performed to examine the effects on the radiation field due to the glass vial and the lack of backscatter material due to the nitrogen-filled space. Results for two different formulations of PAG indicated that the influence of the glass and the nitrogen-filled space are negligible. PMID- 11049194 TI - Comments on 'Standard effective doses for proliferative tumours'. PMID- 11049195 TI - Age-dependent decreases of phosphorus and magnesium in human Achilles' tendons. AB - To elucidate changes of human tendons with aging, the authors studied age-related changes of elements in human Achilles' tendons by inductively coupled plasma atomic emission spectrometry. The subjects consisted of seven men and seven women, ranging in age from 61 to 97 yr. It was found that the content of calcium increased progressively with aging in the Achilles' tendons, whereas the contents of phosphorus and magnesium decreased gradually with aging. The previous investigations demonstrated that the content of calcium and phosphorus increased progressively with aging in most, but not all, human tissues, except for the bones. In ligaments, such as the anterior cruciate ligament and the ligament of the head of the femur, which are histologically similar to the Achilles' tendon, it was previously found that both the contents of calcium and phosphorus increased with aging in the ligaments. It should be noted that the content of phosphorus in the Achilles' tendons decreased during the aging process. In addition, it was found that there was a very high direct correlation between phosphorus and magnesium contents in the tendons, but not between calcium and phosphorus contents. PMID- 11049196 TI - Priming with magnesium-deficient media inhibits preadipocyte differentiation via potential upregulation of tumor necrosis factor-alpha. AB - The effect of priming stromal-vascular cells in primary cultures with magnesium deficient (MgD) media on preadipocyte differentiation was studied. Cultures were derived from dorsal subcutaneous fat tissue of young pigs and maintained 3 d in serum-free control or MgD Dulbecco's modified Eagle's medium, 3 d in 10% fetal bovine serum and dexamethasone, and 6 d in insulin. At d 12 of culture, immunocytochemical and glycerophosphate dehydrogenase assays indicated depressed adipocyte differentiation in the MgD groups. Cultures were enriched for preadipocytes up to 50% of total cells. On the third day of treatment with control and MgD medium, total cell lysates were isolated and 50 microg of them were run on two-dimensional gel electrophoresis. The separated proteins from both treatment groups showed similar patterns. However, spots of proteins with predicted molecular weight in the range of 25.8-37.4 kDa and pI of 5.39-5.85 were sixfold denser from the MgD 10 groups than from the controls. These proteins migrate similarly to tumor necrosis factor-alpha (TNF-alpha). The amount of TNF alpha in cell lysates from the MgD group was about 2.35 times greater than controls determined by TNF-alpha-ELISA. It is likely that proteins upregulated by MgD medium are TNF-alpha isoforms. PMID- 11049197 TI - Radiation exposure does not alter metallothionein III isoform expression in mouse brain. AB - The level of metallothionein III mRNA and protein, a brain-specific isoform of metallothionein (MT), was investigated in the brain of MT-I and -II gene knockout (MT-null) mice exposed to 20 Gy whole-body gamma-irradiation. Because MT-null mice did not express MT-I or MT-II isoforms, the total brain MT content in these mice represented the isoform MT-III only. MT-III protein content was determined by a cadmium-binding assay, and the MT mRNA level was measured by reverse transcription followed by polymerase chain reaction (RT-PCR). Both MT-III protein content and mRNA expression in the brains of MT-null mice were not affected by exposure to whole-body irradiation. These results indicate that mouse brain MT III expression is not induced by ionizing radiation. PMID- 11049198 TI - Zinc distribution in the small-intestinal digesta of pigs fed skim milk powder or defatted soybean flour. AB - Zinc distribution in the small-intestinal digesta was studied in pigs fed a skim milk powder (SMP)-based or defatted soybean flour (DSF)-based diet. Ten pigs were fed experimental diets for 30 d. Serum and femoral zinc concentrations were lower in the DSF group than in the SMP group, suggesting the lower zinc availability in the DSF diet than in the SMP diet. In the digesta of small intestine, zinc solubility was higher in the SMP group than in the DSF group. A gel permeation high-performance liquid chromatography showed that the SMP group exhibited four zinc peaks in the chromatogram of the supernatant of digesta. The chromatogram in the DSF group showed four zinc peaks corresponding to those of the SMP group. However, the first zinc peak was smaller and the second peak was larger in the DSF group than in the SMP group. Dietary treatment did not affect the other peaks of zinc. Although the high solubility of zinc in the digesta of SMP group is considered to result in the high availability of zinc, the difference of zinc distribution in the liquid phase of digesta may partly affect zinc availability. PMID- 11049199 TI - Rats retain chromium in tissues following chronic ingestion of drinking water containing hexavalent chromium. AB - Humans have sometimes been exposed to as much as 10 ppm Cr(VI) in drinking water from contaminated wells. The risks to these individuals are not well understood because the digestive tract reduces some of the Cr(VI) to the less bioavailable Cr(III) prior to absorption, and the disposition of the remaining Cr(VI) has not been well studied. We determined tissue Cr concentrations in rats after chronic ingestion of Cr(VI) in drinking water at concentrations relevant to human exposure levels. Adult male and female Fischer 344 rats consumed ad libitum 0, 0.5, 3, or 10 ppm Cr(VI) as K2CrO4 in drinking water for 44 wk. Rats then were given deionized water 4-6 d prior to sample collection. Females given 3 or 10 ppm Cr(VI) consumed more Cr(VI) per unit of body weight than did males. Bone Cr concentrations were significantly elevated in rats that drank 10 ppm Cr(VI). Renal Cr concentrations were significantly elevated in male rats that drank 3 or 10 ppm Cr(VI) and in female rats dosed with 10 ppm Cr(VI). Female rats had elevated liver Cr concentrations after drinking 3 or 10 ppm Cr(VI). Testicular Cr concentrations were slightly elevated in rats that drank 10 ppm Cr(VI). Brain, ovarian, and whole-blood Cr concentrations were below detection limits in all exposure groups. Although tissue Cr accumulation may have resulted from absorption of Cr(III), it is poorly absorbed. Therefore, the increased tissue retention may also have resulted, in part, from increased absorption of Cr(VI) and its subsequent uptake from the systemic circulation. PMID- 11049200 TI - Selenium regulates expression in rat liver of genes for proteins involved in iron metabolism. AB - Suppression subtractive hybridization analysis in our laboratory recently revealed that transferrin mRNA may be elevated in Sedeficient rat liver. In this work, we compared expression in rat liver of genes for transferrin, transferrin receptor, ferritin light and heavy chains, and iron-regulatory proteins 1 and 2 in Se adequacy and deficiency. Weanling male Sprague-Dawley rats were fed Torula yeast diets supplemented with 0 or 0.15 microg Se/kg diet as sodium selenite for 15 wk. Activity of cellular glutathione peroxidase was virtually abolished in Se deficient rat liver, whereas activity of glutathione S-transferase was 43% higher than in Se-adequate liver. There were no differences in hematocrit, hemoglobin, or liver iron content. To examine differential gene expression, we used a multiplex relative reverse transcriptase-polymerase chain reaction method. Three of the six genes examined showed modest but consistent upregulation in Se deficiency. Transferrin mRNA was 30% more abundant in Se-deficient than in Se adequate liver. For the transferrin receptor, the difference was 32%, and for iron regulatory protein 1, it was 63%. No consistent differences were observed for iron regulatory protein 2 or for ferritin light or heavy chain. These findings suggest a possible role for dietary Se in moderating iron metabolism. PMID- 11049201 TI - Analysis of the phospholipid hydroperoxide glutathione peroxidase mRNA in the rat spermatozoon and effect of selenium deficiency on the mRNA. AB - Phospholipid hydroperoxide glutathione peroxidase (PHGPx) is a selenium (Se) dependent glutathione peroxidase. It is reported that the relative PHGPx mRNA levels are much higher in the testis than in the other tissues. We have analyzed the existence and structure of the PHGPx mRNA in rat sperm and the changes in the level of the PHGPx mRNA after feeding with Se-deficient diets. We used 8-wk-old male Wistar strain rats given Se-adequate feed (control group, n = 5) and Se deficient diets with marginal levels of Se (0.03 ppm or less) (Se-deficient group, n = 5) for 4 wk. The existence and level of the PHGPx mRNA in the cauda epididymal sperm, testis, and liver from the Se-adequate rats were analyzed by the reverse transcription-polymerase chain reaction and the Southern blotting method. As a result, the existence of the PHGPx mRNA was demonstrated in the cauda epididymal sperm as well as in the testis and liver. Moreover, the subtype of the PHGPx mRNA in the rat sperm was the mitochondrial-type mRNA, which included a region corresponding to the mitochondrial transfer leader sequence. These results imply that the intracellular localization of PHGPx may be regulated by the transcription level. On the other hand, there was no significant difference between the control group and the Se-deficient group in the Se level of the cauda epididymal sperm and the level of the PHGPx mRNA. In conclusion, it has been demonstrated that the PHGPx mRNA exists in rat sperm for the first time. The analysis of the PHGPx mRNA in the sperm would be a useful tool for investigating the disfunction caused by the disorder of the level or structure of the PHGPx in the sperm. PMID- 11049202 TI - Time course of arsenite-induced copper accumulation in rat kidney. AB - Oral administration of inorganic arsenic has been shown to lead to an accumulation of copper in the kidneys of rats and guinea pigs. However, nothing is known about the characteristics and mechanisms of this organ-specific renal copper accumulation. Many heavy metals accumulate in the kidney, either after environmental or occupational exposure. An additional accumulation of any other trace metals, even essential ones, may therefore be critical for that organ. This prompted us to follow the course of the renal copper accumulation. Rats were given daily subcutaneous doses of sodium arsenite for 12 d. Each second day, three rats were killed by exsanguination and the liver, kidneys, and blood removed and analysed for As, Cu, and other trace elements by atomic emission spectrometry. Results indicate that arsenic and copper accumulate in the kidney cortex synchroneously over time. Arsenic also accumulated in the liver and red blood cells (RBC). Copper levels in the RBC and liver as well as copper excretion into the urine were unaffected. After terminating arsenite administration, there was a slow decline in tissue levels of both arsenic and copper, a phenomenon which was parallel for both metals. Because the copper level in the liver was not affected, it is concluded from this study that renal processes and not hepatic or biliary mechanisms might be responsible for the renal copper accumulation. Furthermore, the strong linear correlation (r = 0.85) between arsenic and copper levels in the kidney during and after arsenite administration suggests a functional relationship between arsenic and copper with respect to their retention in the kidney. PMID- 11049203 TI - Selenium and immunocompetence in patients with head and neck cancer. AB - This randomized double-blind placebo-controlled study aimed to determine whether oral intake of 200 microg/d of sodium selenite, a dose within the safe and adequate daily intake (50-200 microg/d) recommended by the U.S. Food and Nutrition Board, will abrogate depressed or enhance normal-level immune functions of patients receiving therapy for squamous cell carcinoma of the head and neck. Subjects were given one selenium/placebo tablet/d for 8 wk, beginning on the day of their first treatment for the disease (e.g., surgery, radiation, or surgery and radiation) and their immune functions were monitored. Supplementation with selenium (Se) during therapy resulted in a significantly enhanced cell-mediated immunue responsiveness, as reflected in the ability of the patient's lymphocytes to respond to stimulation with mitogen, to generate cytotoxic lymphocytes, and to destroy tumor cells. The enhanced responsiveness was evident during therapy and following conclusion of therapy. In contrast, patients in the placebo arm of the study showed a decline in immune responsiveness during therapy, which was followed, in some patients, by an enhancement, but the responses of the group remained significantly lower than baseline values. The data also show that at baseline, patients entered in the study had significantly lower plasma Se levels than healthy individuals, and patients in stage I or II of disease had significantly higher plasma selenium levels than patients in stage III or IV of disease. PMID- 11049204 TI - Selenium supplementation of children in a selenium-deficient area in China: blood selenium levels and glutathione peroxidase activities. AB - Keshan disease is a cardiomyopathy restricted to the endemic areas of China and seen in residents having an extremely low selenium (Se) status. Prophylactic administration of sodium selenite has been shown to decrease significantly the incidence of acute and subacute cases. The aim offthe study was to assess the relative bioavailability of selenite versus organic Se-yeast in a Se-deficient area in China with a randomized double-blind double-dummy design. Healthy children (n=30) between 14 and 16 yr of age were randomized into three equal groups receiving either 200 microg/d selenite Se or 200 microg/d Se-yeast or placebo for 12 wk. Blood was drawn at baseline, 4, 8, and 12 wk and 4 wk postsupplementation. The plasma Se concentration (mean +/- SD) was 0.16+/-0.03 micromol/L at baseline. Selenite and Se-yeast supplementation increased plasma Se to plateau values, 1.0+/-0.2 and 1.3+/-0.2 micromol/L, respectively. In red cells, Se-yeast increased the selenium level sixfold and selenite threefold compared to placebo. The relative bioavailability of Se-yeast versus selenite measured as glutathione peroxidase (GSHPx) activity was similar in plasma, red blood cells, and platelets. GSHPx activity reached maximal levels in plasma and platelets of 300% and 200%, respectively, after 8 wk compared to the placebo group, but continued to increase in red cells for 16 wk. Our study showed that although both forms of Se were equally effective in raising GSHPx activity, Se yeast provided a longer lasting body pool of Se. Se-yeast may be a better alternative to selenite in the prophylaxis of Keshan disease with respect to building up of body stores. PMID- 11049205 TI - Zinc uptake by human placental microvillous membrane vesicles: effects of gestational age and maternal serum zinc levels. AB - Zinc uptake by syncytiotrophoblast microvillous membrane vesicles (SMMV) from human placentas was characterized and the effects of maternal serum zinc levels at term and of gestational age on kinetic parameters were evaluated. Zinc uptake at pH 7.2 was rapid for the first 2 min, followed by a slower increase, approaching equilibrium after 30 min. Uptake was saturable at a zinc concentration of 30 micromol/L, higher than the upper range of the physiological serum zinc level. Kinetic analysis of uptake at 1 min in SMMV from term placenta showed similar Km values (mean: 6.9+/-0.6 micromol/L) for different levels of maternal serum zinc. However, Vmax was higher (p < 0.05) in SMMV from mothers with serum zinc lower than 7.6 micromol/L compared to those with higher serum zinc levels (35.8+/-1.6 and 26.6+/-1.6 nmol 65Zn/mg protein/min, respectively). Km values were similar in term (>37 wk of gestation) and preterm (20-25 wk of gestation) placentas, whereas Vmax was higher (p < 0.05) in the preterm (34.3+/ 1.6 nmol Zn/mg protein/min) compared to term placentas from mothers with serum zinc levels above 7.6 micromol/L. These results suggest that whereas afffinity for zinc was not altered with gestational age or maternal serum zinc levels, zinc uptake capacity in human placenta is influenced both by gestational age and by low levels of maternal serum zinc in order to ensure an adequate maternal-fetal zinc transfer. PMID- 11049206 TI - Zinc serum level in human immunodeficiency virus-infected patients in relation to immunological status. AB - In human immunodeficiency virus (HIV) infection, serum level of zinc, an important micronutrient for immune function, is frequently diminished. The aim of this study was to determine the zinc status in relation to immunological parameters and disease stage in 79 HIV-1 seropositive patients. The median serum level of zinc was within normal limits (12.5 micromol/L) but in 23% of patients, zinc deficiency was seen. Decreased serum zinc was associated with a low CD4 cell count, high viral load, and increased neopterin and IgA levels. According to current treatment recommendations, the majority of patients received antiretroviral triple therapy. Zinc levels in treated and untreated patients were comparable. Referring to disease stage (CDC classification, 1993), the mean zinc level was highest in stage C and lowest in stage A. In conclusion, even under antiretroviral triple therapy, zinc deficiency is still of great importance in HIV infection, and zinc substitution in zinc deficient individuals should be taken into account to optimize therapeutical success. PMID- 11049207 TI - Histological change after interferon therapy in chronic hepatitis C in view of iron deposition in the liver. AB - We examined the efficacy of interferon (IFN) therapy for chronic hepatitis C (CHC) in view of the change of liver histology and iron staining before and after IFN therapy. Enrolled in this study were 109 patients with CHC who completed IFN treatment and were followed for at least 1 yr after the end of IFN therapy. Serum iron, unsaturated-iron-binding capacity (UIBC), and total-iron-binding capacity (TIBC) were assessed before IFN therapy. Knodell's histological activity index (HAI) score and iron staining were examined in 55 patients in whom liver biopsy was performed at two points: before and. 1 yr after IFN therapy. Serum iron levels before IFN therapy did not correlate with the response to IFN. The HAI score significantly decreased after IFN therapy in complete responders (p < 0.01) and biochemical responders (p < 0.01). Three factors in the HAI, periportal necrosis, intralobular necrosis, and portal inflammation, but not fibrosis, were significantly decreased in complete responders (p < 0.01) and biochemical responders (p < 0.01). Of 55 patients, 23 (41.8%) were positive for iron staining before IFN therapy and 14 of 55 (25.5%) after IFN therapy. The positive rate for iron staining tended to decrease after IFN therapy, not correlating to the response to IFN, but the change was not statistically significant. In conclusion, the histological improvement by IFN therapy was mostly seen in necroinflammatory changes but not in fibrosis at least 1 yr after IFN, and iron staining tended to decrease after IFN therapy. PMID- 11049208 TI - Atrial natriuretic peptide and thyroid hormones' relation to plasma and heart calcium and magnesium concentrations of Wistar rats exposed to cold and hot ambients. AB - Plasma ANP (atrial natriuretic peptide), thyroid hormones, and calcium and magnesium levels as well as heart tissue calcium and magnesium concentrations were determined in male Wistar rats after exposure of 114 rats at low temperature (4 degrees C) and 95 rats at high temperature (35-36 degrees C) for 28 d. Plasma ANP, triiodothyronine (T3), thyroxine (T4), thyroid-stimulating hormone (TSH), free T3, and free T4 were estimated by radioimmunoassay, and plasma and heart tissue levels of Ca and Mg by flame atomic absorption spectrophotometry. Results were compared to a control group exposed at 20-22 degrees C (76 rats). All the above parameters in control rats did not show statistically significant variations during the study. A significant increase of plasma ANP, T3, T4, Ca, and Mg concentrations developed during cold exposure, whereas a gradual decrease of plasma ANP, T3, T4, and Mg concentrations was revealed during hot exposure. A significant increase of heart tissue Mg concentrations developed during hot exposure. Results also indicate that plasma ANP and T3 levels are proportionally related, whereas an inverse relationship exists between plasma ANP and T3 levels and heart Mg concentrations, in both cold and hot exposed rats. In conclusion, ANP and thyroid hormones in relation to Ca and Mg play an important role in temperature adaptation. PMID- 11049209 TI - Effect of nickel on testicular nucleic acid concentrations of rats on protein restriction. AB - The nucleic acids (DNA and RNA) and total protein concentration in testes were estimated in male Wistar strain rats treated intraperitorally with nickel sulfate (2.0 mg/100 g body weight) on alternate days for 10 dosages. In both normal (18% casein) and protein-restricted (5% casein) experimental animals, the nucleic acids and total protein concentration were found to decrease significantly compared to the corresponding controls. Sperm count and sperm motility were also reduced in both experimental groups of animals. The results indicate that nickel influences the expression of genetic information by reducing testicular nucleic acids and protein concentration in both dietary experimental groups. PMID- 11049210 TI - Plasma and erythrocyte lipid peroxidation levels in patients with testis tumor after orchiectomy. AB - Plasma and erythrocyte lipid peroxidation levels of 20 patients with histopathologically confirmed testis cancer and 20 healthy control individuals were studied between November 1995 and June 1997. The group with testis cancer had a mean age of 24.8+/-8.2 yr and the control group's mean age was 28.3+/-6.9 yr. Stage distribution of the testis cancer cases were 4 of stage A, 10 of stage B, and 6 of stage C. Blood samples of the patients were drawn after orchiectomy and after 12 h fasting before chemotherapy. Mean plasma and erythrocyte lipid peroxidation levels were detected to be 14.51+/-5.30 nmol malondialdehide (MDA)/mL and 9.30+/-2.06 nmol MDA/g hemoglobin (Hb), respectively, in the testis cancer group, whereas the healthy control group had mean plasma and erythrocyte lipid peroxidation levels of 10.7+/-1.82 nmol MDA/mL and 6.18+/-1.68 nmol MDA/g Hb, respectively. Plasma and erythrocyte lipid peroxidation values of the testis cancer patients were determined to be statistically significantly higher than that of the health control group (p < 0.001, p < 0.001). No significant correlation was determined between plasma, erythrocyte lipid peroxidation levels and tumor markers. In conclusion, it can be said that an increase in the lipid peroxidation may play a role in the pathogenesis of testis carcinomas in addition to the other causes. PMID- 11049211 TI - Alteration in cardiomyocyte mechanics by suboptimal levels of extracellular magnesium. AB - The beneficial effects of magnesium supplementation in pathological situations is well known, but the myocardial response to a nominal decrease in the level of magnesium has received relatively little attention. Hypomagnesemia can occur as chronic or acute manifestation of physiological changes, pathological conditions, or pharmacological interventions. Experimental interest was focused on the mechanical changes in adult rat heart myocytes following variation in extracellular Mg2+. Isolated cells were exposed to different levels of extracellular Mg2+ and the amplitude and rate of contraction were measured as a function of change in cell length using a video-based edge-detection system. Investigations have revealed that variation in the level of Mg2+ within physiological limits leads to mechanical changes. A decrease in the level of extracellular Mg2+ was accompanied by a significant increase in contractile amplitude and decrease in the velocities of contraction and relaxation. The contractile amplitude measured as percentage shortening were 3.08+/-0.19%, 4.62+/ 0.19% and 6.9+/-0.40%, respectively, on exposure to 1.8, 0.8, and 0.48 mM Mg, and the corresponding velocities of contraction and relaxation normalized to amplitude were 0.54+/-0.02, 0.40+/-0.03, 0.31+/-0.03 and 0.47+/-0.02, 0.35+/ 0.02, 0.24+/-0.02. The variations in contractile parameters associated with the change in the level of Mg were statistically significant (p < 0.01). Variation in the contractile properties associated with change in extracellular Mg2+ may be effected by alteration in Ca2+ transients. PMID- 11049212 TI - Determination of selenium in serum by FI-HG-AAS and calculation of dietary intake. AB - A method was developed for the determination of selenium concentration in serum by flow injection-hydride generation-atomic absorption spectrometry (FI-HG-AAS) following microwave digestion of serum samples and reduction of selenate to selenite. The detection limit of the method was 0.3 microg Se/L and the characteristic concentration, corresponding to the 0.0044 absorbance signal, was 0.12 microg Se/L. The results from the analysis of two Seronorm standard reference materials showed good agreement with the certified values. The method was then used to analyze selenium in sera of Austrian and Slovenian people for the calculation of dietary intakes. The selenium concentrations in sera of mothers at delivery, their neonates, and the male and female adults were 71+/-14, 42+/-6, 75+/-21, and 65+/-16 microg/L for the Austrians and 62+/-15, 34+/-7, 70+/ 12, and 66+/-15 microg/L for the Slovenians. The dietary intakes of selenium of the mothers and the male and the female adults were calculated as 52, 37, and 46 microg/d for the Austrians and 45, 38, and 32 microg/d for the Slovenians. PMID- 11049213 TI - Serum, urinary, and fecal calcium changes in trained and untrained subjects during prolonged hypokinetic and ambulatory conditions. AB - Electrolyte metabolism undergoes significant changes in trained subjects, but it is unknown if it undergoes significant changes in untrained subjects during hypokinesia (decreased movement). The aim of this study was to measure calcium (Ca) changes in trained and untrained subjects during prolonged hypokinesia (HK). Studies were done during 30 d of a pre-HK period and 364 d of a HK period. Forty male trained and untrained volunteers aged 23-26 yr were chosen as subjects. All subjects were equally divided into four groups: trained ambulatory control subjects (TACS), trained hypokinetic subjects (THKS), untrained hypokinetic subjects (UHKS), and untrained ambulatory control subjects (UACS). The THKS and UHKS groups were kept under an average running distance of 0.7 km/d. Fecal Ca excretion, urinary Ca and magnesium (Mg) excretion, serum ionized calcium (CaI), Ca, Mg, intact parathyroid hormone (iPTH) and 1,25 dihydroxyvitamin D [1,25 (OH)2 D] concentration, body weight, and peak oxygen uptake were measured. Fecal Ca loss, urinary Ca and Mg excretion, and serum CaI, Mg, and Ca increased significantly (p < or = 0.01), whereas serum iPTH and 1,25 (OH)2 D concentration body weight and peak oxygen uptake decreased significantly (p < or = 0.01) in the THKS and UHKS groups when compared with the TACS and UACS groups. The measured parameters were much greater and much faster in the THKS group than in the UHKS group. By contrast, the corresponding parameters did not change significantly in the TACS and UACS groups when compared with the baseline control values. It was concluded that prolonged HK induces significant fecal, urinary, and serum Ca changes in the hypokinetic subjects when compared with control subjects. However, fecal, urinary, and serum Ca changes were much greater and appeared much faster in the THKS group than the UHKS group. PMID- 11049214 TI - Calcium supplementation effect on calcium balance in endurance-trained athletes during prolonged hypokinesia and ambulatory conditions. AB - Calcium (Ca) supplements may be used to normalize Ca-balance changes but little is known about the effect of Ca supplements on Ca balance during hypokinesia (decreased kilometers per day). The aim of this study was to evaluate the effect of daily intakes of Ca supplements on Ca balance during hypokinesia (HK). Studies were done during 30 d of a pre-HK period and during 364 d of a HK period. Forty male athletes aged 23-26 yr were chosen as subjects. They were divided equally into four groups: unsupplemented ambulatory control subjects (UACS), unsupplemented hypokinetic subjects (UHKS), supplemented hypokinetic subjects (SHKS), and supplemented ambulatory control subjects (SACS). The SHKS and UHKS groups were kept under an average running distance of 0.7 km/d. In the SHKS and SACS groups supplemented with 35.0 mg Ca lactate/kg body weight. Fecal Ca loss, urinary excretion of Ca and phosphate (P), serum concentrations of ionized calcium (CaI) total Ca, P, and Ca balance, intact parathyroid hormone (iPTH) and 1,25 dihydroxyvitamin D (1,25(OH)2D), anthropometric characteristics and peak oxygen uptake were measured. Fecal Ca excretion, urinary Ca and P excretion, serum CaI, total Ca, and P concentration, and negative Ca balanced increased significantly (p < or = 0.01) in the SHKS and UHKS groups when compared with the SACS and UACS groups. Serum, urinary, and fecal Ca changes were much greater and appeared much faster in the SHKS group than in the UHKS group. Serum iPTH and 1,25 (OH)2 D, body weight, and peak oxygen uptake decreased significantly (p < or = 0.01) in the SHKS and UHKS groups when compared with the SACS and UACS groups. In contrast, the corresponding parameters remained stable in the SACS and UACS groups when compared with the baseline control values. It was concluded that during prolonged HK, urinary and fecal Ca excretion and serum Ca concentration increased significantly despite the presence of a negative Ca balance; thus, Ca supplements cannot be used to normalize negative Ca balance during prolonged HK. PMID- 11049215 TI - Natural polyphenols-iron interaction: its biological importance. AB - The iron-binding capacity of different fractions of natural polyphenols extracts was determined by chromatographic and electrophoretic methods. Their effects on iron-induced calcium homeostasis changes in liver tissue suspension showed that mate tea and green tea extracts provoke a very significant inhibition of the iron effects, whereas it is much less significant with red wine extract. The biological importance of this phenomenon is discussed. PMID- 11049216 TI - Iron distribution in different tissues in rats following exercise. AB - Iron plays an essential role in blood oxygen transport and in muscle physiology. No conclusive data exist in the literature concerning its tissue distribution and behavior following exercise and training. The aim of the present work was to analyze the Fe content in different tissues following a single session of swimming to exhaustion and after swimming training in rats in order to more extensively describe the changes of Fe distribution provoked by exercise. Animals were divided into four groups (n=10): control group at rest, trained group at rest, control group after exercise, and trained group after exercise. First, rats swam until exhaustion and the maximal swimming time was noted. The training protocol consisted of swimming (5 d/week for 3 wk), limiting the time to 60% of the maximum obtained during the first session to exhaustion of each rat. The variables measured were erythrocytes, hemoglobin, hematocrit, mean corpuscular volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and, Fe in liver, kidney, erythrocytes, heart, muscle, bone, and serum. Variations in plasma volume were also calculated. Tissues presented two different profiles with regard to the changes of Fe concentration provoked by training: those displaying higher values of Fe after training, such as liver, heart, muscle, and serum, and those displaying lower values, such as bone, kidney, and red blood cells. These changes in the distribution of Fe in different tissues could be the result of an increase in the needs and use of Fe, shown by active tissues at exercise, and it is possible that the hormonal changes provoked by stress lead to a different behavior of Fe proteins. PMID- 11049217 TI - Cytochrome c oxidase, Cu,Zn-superoxide dismutase, and ceruloplasmin activities in copper-deficient bovines. AB - The activity of several cuproenzymes in relation to the immune system was examined in serum and blood cells from bovines with molybdenum-induced copper deficiency. Five female cattle were given molybdenum (30 ppm) and sulfate (225 ppm) to induce experimental secondary copper deficiency. Ceruloplasmin activity was determined in serum. The Cu,Zn-superoxide dismutase and cytochrome c oxidase activities were measured in peripheral blood lymphocytes, neutrophils, and monocyte-derived macrophages. Copper deficiency was confirmed from decreased serum copper levels and the animals with values less than 5.6 micromol/L were considered deficient. The content of intracellular copper decreased between 40% and 70% in deficient cells compared with the controls. In copper-deficient animals, the serum ceruloplasmin activity decreased to half of the control value. Both of them, the Cu,Zn-superoxide dismutase and the cytochrome c oxidase activities, undergo a significant reduction in leukocytes, showing differences among diverse cell populations. We concluded that the copper deficiency alters the activity of several enzymes, which mediate antioxidant defenses and ATP formation. These effects may impair the cell immune functionality, affecting the bactericidal capacity and making the animals more susceptible to infection. PMID- 11049218 TI - Zinc status and immune system relationship: a review. AB - The essentiality of zinc for humans was first documented by Prasad in the 1960s. The main clinical manifestations associated with zinc deficiency are growth retardation, hypogonadism, diarrhea, and increased susceptibility to infectious diseases. Thus, in the past 25 yr, there was an increased interest of researchers in studying the role of zinc in human immunity. Although mechanistic research has been carried out using animal models, there are several studies in humans with similar results. This work is an attempt to review the information available in this field to understand the important role that zinc plays in the normal development and function of the immune system. PMID- 11049219 TI - Trace elements and lipid peroxidation in human seminal plasma. AB - In the present study, the concentrations of copper, iron, zinc, and malondialdehyde in human seminal plasma were measured and correlated with the sperm count and motility in human semen. Copper, iron, and zinc were analyzed by atomic absorption spectrometry, whereas malondialdehyde was measured by high performance liquid chromatography. The malondialdehyde concentrations in asthenospermia and oligoasthenospermia were significantly higher than in normospermia. Copper and iron levels were higher in asthenospermia, whereas the zinc concentrations in both oligospermia and asthenospermia were lower than in normal controls. A negative correlation (r = -0.28, p < 0.05) between the malondialdehyde concentration and sperm motility was observed in the abnormal groups. There was no association among copper, iron, zinc, and malondialdehyde in seminal plasma. We concluded that changes in trace elements may be related to sperm quality and that lipid peroxidation, although it is not promoted in the seminal plasma by copper or iron or ameliorated by zinc, may be involved in the loss of sperm motility. PMID- 11049220 TI - A study of factors that may influence the determination of copper, iron, and zinc in human milk during sampling and in sample individuals. AB - The aim of this study was to establish the possible effects of the sampling protocol (between-breast, within-feed, and diurnal differences) and the mother's personal factors (age, parity, iron supplementation, smoking habits, and lactation period) on the copper, iron, and zinc contents in human milk. One hundred thirty-six human milk samples identified by their origin and sampling conditions were analyzed. The samples were obtained from the 2nd to 15th d postpartum from 62 women. The data on the individuals required for the study were available. Mineral determinations were analyzed by flame atomic absorption spectrometry following a standardized protocol. The results showed that iron contents were higher in hind-milk samples and at the nighttime feeding and depended on the breast from which the sample was taken. The copper and zinc concentrations showed no significant variations. There was no significant relationship among the mothers' age, parity, smoking habits, iron supplementation, and copper content. Milk from older women had lower zinc contents than that of younger women. Increased amounts of iron were found in multiparous women. Between colostrum and transitional milk, a sharp decrease in zinc content was observed, whereas copper and iron contents remained constant. All of these results make it clear that standardized sampling protocols are needed in order to obtain comparable values. PMID- 11049221 TI - Effect of zinc on cellular levels of calmodulin and cyclic adenosine monophosphate in the adipocyte. AB - A perturbation of zinc metabolism has been noted in subjects with obesity. Zinc may also participate in the intracellular signal cascade by affecting cellular calcium influx and a change in the calcium-calmodulin (CaM)-cyclic adenosine monophosphate (cAMP) pathway. The possible effects of zinc on cellular concentrations of CaM, a major cytosolic calcium-binding protein, in the adipocytes derived from obese (ob/ob) mice and their lean counterparts were studied. Adipocytes derived from both phenotypes of mice were treated either with 0.2 mM of zinc sulfate or without any additive for 1 h of incubation; the cellular levels of CaM and cAMP were then determined. The results showed that the obese mice had lower CaM and cAMP levels in their adipocytes compared to the lean mice. Zinc treatment reduced CaM and increased cAMP levels in all mice, although this effect was more pronounced in the lean mice. This study indicated that there was an inverse interaction between CaM and cAMP in their cellular levels in the mouse adipocytes and that might be affected by exogenous zinc addition. PMID- 11049222 TI - Determination of trace elements in some natural drugs by atomic absorption spectrometry. AB - Fifteen kinds of common plants, animals, and minerals used as traditional medicines by the Chinese people have been subjected to analysis by atomic absorption spectrometry for its content of seven metals: lead, cadmium, arsenic, mercury, copper, cobalt, and manganese. The concentrations of these elements are significantly different according to their vegetal, animal, or mineral origin. The average values found for lead, cadmium, arsenic, cobalt, and manganese in drugs of mineral origin are higher than those derived from plants and animals, except for copper, which was higher in drugs of animal origin. Our results suggest that the user of traditional Chinese crude drugs should be warned of the potential danger of heavy-metal poisoning because their concentrations seem to be higher than the maximum values allowed by health agencies in several countries. PMID- 11049223 TI - Effects of acute and chronic coingestion of AlCl3 with citrate or polyphenolic acids on tissue retention and distribution of aluminum in rats. AB - Aluminum (Al) is toxic to certain biological systems and has been implicated as a neurotoxic agent in the pathogenesis of Alzheimer's disease. Intestinal absorption of Al is very low (0.1%), but many organic dietary components are potential chelators of Al and may enhance its absorption and tissue distribution. We examined the effects of acute and chronic coingestion of AlCl3 with different polyphenolic acids on Al retention and compared to citrate in rats. In experiment 1, animals fasted for 14 h were dosed orally with demineralized water, Al chloride, Al chloride plus sodium citrate, or Al chloride plus a polyphenol acid. Blood samples were taken before and 2 h after the gavage and animals were killed 6 h later. In experiment 2, the rats were adapted on a purified diet for 1 wk and received the following for 4 wk in their experimental diets: AlCl3, except group 1, plus citrate or a polyphenol acid, except groups 1 and 2. Animals were killed and blood and tissues were sampled. In experiment 1, citrate highly enhanced Al absorption and its tissue retention. Gallic and chlorogenic acids significantly increased tibia and kidney Al levels compared to the Al group. In experiment 2, Al levels in the urine were significantly increased in all the Al groups compared to the control group. Significantly higher Al levels in the tibia, kidney, and brain were observed in the citrate group and a significant increase in brain Al level was also noted in the chlorogenic acid group compared to AlCl3 group. This may suggest a possible relation structure-activity of polyphenol acids. However, further studies are necessary to better understand the influence of polyphenol acids on Al metabolism, in particular that of chlorogenic acid. PMID- 11049224 TI - Effects of cadmium administration on the endogenous metal balance in rats. AB - The concentrations of cadmium and other metal ions in selected organs, urine, and blood of female rats were measured after exposure to cadmium chloride through their diet or by oral or intravenous administration. The hematological and urinary variations were followed for 4 wk. Body weight gain and the weights of livers and kidneys from all treated groups were not significantly different from the controls. No gross morphological changes were observed in any of the tissues studied at necropsy. The accumulation of cadmium occurred in the liver and kidney. The zinc levels in these organs were elevated relative to controls, in all treated groups regardless of dose and exposure route. Copper was elevated in the liver, kidney, bone, and blood of animals subject to intravenous administration of cadmium. Hepatic iron was decreased in the dietary and orally treated groups, but was not affected in the intravenous study group. The level of magnesium in kidney was increased for all exposure routes, but that of liver was increased only in the intravenously injected groups. The changes in the concentrations of sodium, potassium, calcium, and phosphorus did not follow a specific pattern and varied from organ to organ, depending on the exposure route. The discussion includes a relationship between tissue injury and the alteration of tissue essential element concentrations. PMID- 11049225 TI - A SXRF method for determining the relative concentration of trace elements in plasma protein affected by cisplatin. AB - This article presents an analytical method for the determination of the relative concentrations of trace elements in plasma protein by gel chromatography combined with SXRF (synchrotron radiation X-ray fluorescence). The fraction of plasma protein of male Kunming mice (body weight 24.2 +/- 0.3 g), treated with a cisplatin i.p. injection at a dose of 10 mg/kg, was obtained by the separation of a Sephadex G-50 column (buffered with ammonium acetate, pH 5.7). The SXRF experiments were performed at the Beijing Electron Positron Collider synchrotron radiation facility. The elements (Pt, S, Ca, Fe, Ni, Cu, Zn, Se, Br, and Sr) in the fraction of the plasma proteins (>22 kDa) were assayed using highly sensitive SXRF. The relative concentrations of elements were calculated by a normalization of Compton scattering intensity around 22 keV, after the normalization for the collection time of the X-ray spectrum and the counting of the ion chamber, and subtracting the contribution of the polycarbonate film for the supporting sample. The determination could prove that the element Pt in plasma was bound with macromolecular protein. Cu and S were present in the fraction of the protein in mice treated with cisplatin increase, and their ratios of treated/control were 1.66 +/- 0.06 and 1.78 +/- 0.33, respectively; Zn decreased to a ratio of 0.78 +/ 0.09. Our results are in agreement with others that cisplatin exposure leads to a marked loss of kidney copper and a moderate rise in kidney zinc. However, this article primarily describes one of the analytical methods used; it does not emphasize the results of the effect of cisplatin on trace elements in plasma protein. PMID- 11049226 TI - Concentrations of selected trace elements in human milk and in infant formulas determined by magnetic sector field inductively coupled plasma-mass spectrometry. AB - Magnetic sector field inductively coupled plasma-mass spectrometry (ICP-MS) was applied to the reliable determination of the 8 essential trace elements cobalt (Co), chromium (Cr), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), selenium (Se), and vanadium (V) as well as the 7 nonessential and toxic elements silver (Ag), aluminum (Al), arsenic (As), gold (Au), platinum (Pt), scandium (Sc), and titanum (Ti) in 27 transitory and mature human milk samples and in 4 selected infant formulas. This advanced instrumentation can separate spectral overlaps from the analyte signal hampering significantly the determination of many trace elements by conventional ICP-MS. Moreover, superior detection limits in the picogram per liter range can be obtained with such magnetic sector field instruments. Therefore, this is the first study to report, the concentrations of the elements Ag, Au, Pt, Sc, Ti, and V in human milk and in infant formulas. Concentrations of Ag (median: 0.41 microg/L; range: < 0.13-42 microg/L) and Au (median: 0.29 microg/L; range 0.10-2.06 microg/L) showed large variations in human milk that might be associated with dental fillings and jewelry. Pt concentrations were very low with most of the samples below the method detection limit of 0.01 microg/L. Human milk concentrations of Co (median: 0.19 microg/L), Fe (380 microg/L), Mn (6.3 microg/L), Ni (0.79 microg/L), and Se (17 microg/L) were at the low end of the corresponding reference ranges. Concentrations of Cr (24.3 microg/L) in human milk were five times higher than the high end of the reference range. For Al (67 microg/L), As (6.7 microg/L), and V (0.18 microg/L), most of the samples had concentrations well within the reference ranges. All elemental concentrations in infant formulas (except for Cr) were approximately one order of magnitude higher than in human milk. PMID- 11049227 TI - Calcium measurements in primates during and after hypokinesia in establishing calcium deficiency during prolonged hypokinesia. AB - Hypokinesia (diminished movement) induces significant calcium (Ca) changes, but little is known about the effect of hypokinesia (HK) on Ca deficiency. Measuring Ca changes during and after HK the aim of this study was to determine Ca deficiency during prolonged HK. Studies were done on 12 male Macaca mulatta (rhesus monkeys) aged 3-5 yr (5.58-6.42 kg) during a 90-d pre-HK period, a 90-d HK period, and a 15-d post-HK period. Monkeys were equally divided into two groups: vivarium control monkeys (VCM) and hypokinetic monkeys (HKM). Hypokinetic monkeys were kept in small individual cages that restricted their movements in all directions without hindering food and water intakes. Urinary, fecal, and serum Ca, urinary and serum magnesium (Mg) and phosphate (P), serum intact parathyroid hormone (iPTH), and calcitonin (CT) concentration, body weight, food intake, fluid consumed and eliminated in urine were measured. During the HK period, fecal Ca loss, urinary Ca, P, and Mg excretion, fluid elimination, and serum P, Ca, and Mg concentration increased significantly (p < or = 0.01), whereas serum iPTH and CT concentration, food and fluid intakes, and body weight decreased significantly (p < or = 0.01) in the HKM group when compared with the VCM group. During the initial days of the post-HK period, serum Ca, Mg, and P concentration, fecal Ca loss, urinary Ca, Mg, and P excretion, and fluid elimination decreased significantly (p < or = 0.01), whereas fluid intake increased significantly (p < or = 0.01) in the HKM group when compared with the VCM group. Food intake, body weight, and serum iPTH and CT concentrations remained significantly (p < or = 0.01) depressed in the HKP group when compared with the VCM; however, they increased as the duration of the post-HK period increased. By contrast, the corresponding parameters remained stable in the VCM group when compared with the baseline control values. It was shown that fecal and urinary Ca loss and serum Ca concentration increases significantly during HK, whereas during postHK fecal, urinary, and serum Ca decreases significantly. It was concluded that significant decrease of serum, urinary, and fecal Ca during post-HK may suggest the presence of Ca deficiency during prolonged HK. PMID- 11049228 TI - Effects of EDTA on trace elements and cardiovascular function in the anesthetised rabbit. AB - The present study was undertaken in order to study the effects of the broad acting chelating agent CaNa2-EDTA on plasma trace elements and cardiovascular function in anesthetised New Zealand White rabbits. Trace elements are important for cardiovascular and immune functions and the rabbit is a well-accepted species in cardiovascular studies. The test compound CaNa2-EDTA was administered intravenously to rabbits at single doses of 4, 20, and 100 mg/kg. In addition, at 20 mg/kg, the effects of a second dose after 3 h were also investigated. Heart rate, blood pressure and body temperature were continuously monitored during a 6 h interval after injection of CaNa2-EDTA. Immediately before administration (-1 min) and at 3 and 6 h over the period of the experiment, the plasma cytokine response (tumor necrosis factor-alpha) and trace elements (Mn, Cu, Zn, Se, Ag, Cd, Hg, Pb) were measured. Regardless of dose, blood pressure was found to decrease, but no corresponding changes in heart rate were observed. Both a repeated administration of 20 mg/kg and a single dose of 100 mg/kg were detrimental and caused severe cardiovascular effects and lethality. alpha-TNF tended to increase, though only at 100 mg/kg. The electrocardiogram and body temperature were not affected by the treatment. The most pronounced trace element change was a dose-dependent increase in Mn that was equally pronounced at all time-points after the dose. There was an initial decrease in Cd at low dose levels (4 and 20 mg/kg) that turned into an increase after 6 h at 20 mg/kg and from 2 h at 100 mg/kg. A similar pattern with pronounced decreases at low dose levels was observed for Zn. Cu decreased similarly at all dose levels. For the other trace elements, no or inconsistent effects were observed. This model allows the study of concomitant cardiovascular and trace element changes during treatment with drugs and chelating agents preceding a possible lethal end point and associated pathophysiologic changes. PMID- 11049229 TI - Trace element changes in the myocardium during coxsackievirus B3 myocarditis in the mouse. AB - During most infections plasma, concentrations of trace elements change, but it is unclear if this reflects changes in infected target tissues. In coxsackievirus B3 (CB3) infection, the myocardium is a target in both humans and mice. The concentrations of 12 trace elements were analyzed by inductively coupled plasma mass spectrometry (ICP-MS) in the myocardium of sham-inoculated controls and infected A/J mice 4 and 7 d postinoculation. The size of the inflammatory lesion was positively correlated to the virus content of the heart, as estimated by histopathology and in situ hybridization, respectively. Iron, cobalt, vanadium, and selenium showed transient changes, whereas for the other elements, tendencies on d 4 were manifest on d 7. A three-fold increase in calcium on d 7 suggests prestages of calcification, whereas increases in zinc, selenium, and copper may be the result of the accumulation of immune cells. The magnesium decrease may contribute to the increased sensitivity to cardiac arrhythmias in myocarditis. PMID- 11049230 TI - Chemical forms of selenium present in rat and ram spermatozoa. AB - In vivo and in vitro studies were conducted to investigate the chemical forms by ion-exchange chromatography of selenium (Se) present in rat and ovine spermatozoa. After injection with 75Se-selenite, the form of 75Se in rat sperm was selenocysteine, but selenocysteine and selenomethionine (SeMet) were present in ovine sperm. Presumably, synthesis of SeMet by rumen microbes are responsible for its presence in ovine sperm. In vitro incubation of ram sperm with selenocysteine or SeMet produced no changes, but incubation with selenite produced a compound that eluted one fraction before SeMet from the ion-exchange column. After treatment of this fraction with mercaptoethanol, it eluted in a later fraction upon rechromatography, suggesting it to be selenodicysteine. This compound is apparently formed because of high levels of cysteine in semen. Cysteine, reduced glutathione, and oxidized glutathione were also found in semen. The significance of the results is discussed. PMID- 11049231 TI - Study on the dose-effect relationship of selenite with the growth of wheat. AB - The wheat in hydroponics culture has been chosen as a model to study the dose effect relationship of selenite with its growth. Five different morphological end points and 12 different biological and biochemical end points during different phases of growth of wheat seedling and seed germination have been measured and analyzed. A dose-effect relationship of selenite with the growth of wheat has been obtained from data analysis. The results shows that critical threshold for beneficial effect is about 1.0 mg Se/L, and the critical threshold for adverse effect is about 5.0 mg Se/L in hydroponics culture. From the result of product of lipid peroxidation, it is assumed that the role of selenium for plant is related to the oxy-radical reaction. PMID- 11049232 TI - Spectroscopic and magnetic properties of a Ni(II) complex with citric acid. AB - The complex compound K2[Ni(cit)(H2O)2]2 x 4H2O (cit = triionized citrate ion) seems to be a good model for the investigation of Ni(II)/citrate interactions that are of interest in relation to nickel metabolism and bioaccumulation. Its infrared and Raman spectra were recorded and analyzed on the basis of its structural peculiarities. The magnetic susceptibility, investigated in the temperature range between 1.9 and 300 K, shows the absence of magnetic interactions between the two metal centers present in this structure. PMID- 11049233 TI - Recent advances in the management of gynecologic cancers. PMID- 11049234 TI - Prevalence of human papillomavirus infection and its correlation with cervical lesions in commercial-sex workers in Japan. AB - OBJECTIVE: To investigate the prevalence of the human papillomavirus (HPV) infection and its correlation with cervical lesions in commercial-sex workers (CSWs) who attended a sexually transmitted disease (STD) clinic in an entertainment area in Tokyo. METHODS: Surveys were conducted on 546 prostitutes and 233 control subjects. In all subjects, HPV detection was performed by the hybrid capture method. A cervical cytological examination was performed on 247 prostitutes and 233 control subjects. RESULTS: The HPV-positive rates in the two periods of study were higher (p < 0.01) in CSWs than in the control subjects. When the cytological grades were examined according to HPV-positive rates, the proportion of cytologic Class IIIa to Class IV was significantly higher (p < 0.01) in the HPV-positive CSWs than in the HPV-negative CSWs or in the normal subjects. CONCLUSION: The high frequencies of HPV infection and cervical dysplasia in the CSWs in the present series might predict a higher risk of cervical cancer in this group of subjects. PMID- 11049235 TI - A clinicopathological study of postoperatively upgraded early squamous-cell carcinoma of the uterine cervix. AB - OBJECTIVE: To investigate the clinicopathological backgrounds and diagnostic problems of postoperatively upgraded early squamous-cell carcinomas of the uterine cervix. PATIENTS AND METHODS: A total of 23 patients with postoperatively upgraded early squamous-cell carcinomas who were treated at the Saitama Cancer Center during the period of January 1, 1976, through December 31, 1991, were analyzed clinicopathologically. We reexamined the Pap smears (ectocervix, endocervix), colposcopic findings, punch biopsies, and histological findings of the operative specimens. All patients were divided into one of 3 groups based on each patient's main location of the carcinoma of the cervix: Type A: ectocervical type; Type B: endocervical type; or Type C: combined (ectocervical and endocervical) type. Clinical staging of the uterine cervical carcinomas was done in accordance with the 1994 FIGO rules. RESULTS: The numbers of patients were: Type A, 2; Type B, 10; Type C, 11. Of the 23 patients, 21 (91.3%) had lesions in the endocervical portion at least. Fifteen patients (65.2%) complained of atypical vaginal bleeding. Colposcopic findings suggesting an invasive carcinoma appeared for only 6 patients (26.1%). A cytological reevaluation revealed that the endocervical findings were much stronger than the ectocervical ones in 10 (66.7%) of 15 patients whose smears of both sites could be rechecked. CONCLUSIONS: Even if the preoperative diagnosis was early cervical carcinoma, CIS or Stage Ia1, the signs of atypical vaginal bleeding suggested that the final clinical stage would be upgraded after an operation. Furthermore, when the endocervical cytological findings were much more exaggerated than the ectocervical ones, the possibility of deeply invaded endocervical lesions should be considered. PMID- 11049236 TI - Effectiveness of second- and third-generation immunoassays for the detection of hepatitis C virus infection in pregnant women. AB - OBJECTIVE: To compare the sensitivity and specificity of a third-generation anti hepatitis C virus (anti-HCV) assay (HCV 3.0) with second-generation one (HCV 2.0), and correlate with HCV-RNA positivity by reverse-transcription polymerase chain reaction (RT-PCR). METHODS: We enrolled 197 pregnant women without screening for alanine aminotransferase (ALT) (Group A) and 324 pregnant women with elevated ALT activity (> 45 IU/l) (Group B). Each serum sample was tested by second- and third-generation tests, and anti-HCV titer was determined by serial dilutions. Anti-HCV-positive samples were subjected to HCV-RNA assays. RESULTS: Three (1.5%) and 4 (2.0%) of the 197 group A subjects were anti-HCV-positive by the second- and third-generation methods, respectively, while 17 (5.3%) and 21 (6.5%) of group B were positive, respectively. Three (1.5%) in group A and 8 (2.5%) in group B were HCV 2.0-negative, but positive for HCV 3.0. The sensitivity, specificity, positive and negative predictive values of HCV 2.0 and HVC 3.0 for positive HCV-RNA in both groups were 67 vs 100%, 30 vs 75%, 67 vs 92%, and 30 vs 100%, respectively. The anti-HCV-positive samples with optical density (O.D.) < 1.0 either by second- or third-generation test were negative for HCV-RNA, whereas samples with O.D. > or = 1.0 were all HCV-RNA-positive. CONCLUSIONS: The performance of HCV 3.0 is better than that of HCV 2.0, and anti HCV-positive samples with O.D. < 1.0 are negative for HCV-RNA. PMID- 11049237 TI - Epidural analgesia in primigravidae in spontaneous labour at term: a prospective study. AB - OBJECTIVES: To prospectively study the intervention rate, duration of labour, malpositions, fetal outcome, maternal satisfaction, voiding complications and adverse events in healthy primigravidae in spontaneous labour at term following epidural analgesia. METHODS: A prospective randomized study involving 55 patients in the epidural group and 68 in the control pethidine--inhalational entonox group. RESULTS: There were significantly more obstetric interventions (instrumental deliveries) in the epidural group (p < 0.01). The total duration of labour and the duration of the second stage was prolonged in the epidural group (p < 0.01). There were more malpositions at the second stage of labour in the epidural group (p < 0.02). There were no differences in fetal outcome (Apgar scores and Special Care Nursery admissions). Patients in the epidural group were consistently happier with their method of pain relief (p < 0.01). Two patients required blood patches while another 2 patients had persistent backache post epidural analgesia. CONCLUSION: Epidural analgesia in primigravidae in spontaneous labour at term led to an increased instrumental delivery rate, prolonged duration of labour, greater rate of malpositions in the second stage, increased oxytocin requirements but with no difference in fetal outcomes but with happier mothers as compared to the control group. PMID- 11049238 TI - Sonohysterography in the diagnosis of abnormal uterine bleeding. AB - OBJECTIVE: The purpose of this study was to evaluate the value of sonohysterography in diagnosis of patients with abnormal uterine bleeding. METHOD: Fifty-five patients with abnormal uterine bleeding and transvaginal sonography suggested an abnormal endometrial echo were enrolled in the study. Sonohysterography was performed on all patients, using saline instilled through an endocervically placed catheter. The histologic or pathologic finding was evaluated after surgical procedures and compared with sonohysterography results. RESULTS: Sonohysterography was successfully completed in 52 cases (94.54%). Three cases had cervical stenosis and failed to pass the catheter into the uterine cavity. Mean age was 41.56 +/- 7.82 years (range 29-58 years). Forty-six of 52 sonohysterography demonstrated intrauterine abnormalities (29 endometrial polyps, 15 submucous myomas, 2 endometrial hyperplasia). The pathologic finding demonstrated 46 intrauterine pathologic cases (28 endometrial polyps, 15 submucous myomas, 3 endometrial hyperplasia). Sonohysterography had 97.82% sensitivity, 83.33% specificity, 97.82% positive predictive value, 83.33% negative predictive value, and 96.15% accuracy. CONCLUSION: Sonohysterography is a highly sensitive, specific, and accurate screening procedure for the evaluation of uterine cavity in abnormal uterine bleeding and is a simple, minimally invasive, and effective tool to use in the evaluation of patients. PMID- 11049239 TI - Uterine leiomyoma causing massive ascites and left pleural effusion with elevated CA 125: a case report. AB - A 51-year-old patient presented with an abdominal mass and ascites as well as a left pleural effusion. Her serum CA125 was 820 U/ml. Surgical exploration revealed a benign leiomyoma of the uterus without malignant cytology in the ascites. Postoperatively, the pleural effusion was resolved dramatically and the CA125 decreased to the normal range after 4 months post-operatively. This is an extremely rare case of pseudo-Meigs' syndrome caused by uterine leiomyoma. PMID- 11049240 TI - Surgical management of genital prolapse: is chain cystourethrography useful for evaluating anatomical corrections and urinary symptoms after surgery? AB - OBJECTIVE: To evaluate the relationship between chain cystourethrography and surgery for genital prolapse and urinary symptoms. METHODS: A retrospective study of women with genital prolapse was conducted from January 1991 through December 1997. To assess the preoperative and postoperative anatomical situations objectively, chain cystourethrography was performed. RESULTS: Fifty-seven patients underwent surgical repairs for genital prolapse. All of them recovered from the genital prolapse both subjectively and objectively after surgery. Chain cystourethrography is useful for evaluating postoperative anatomical corrections. However, 6 patients (12%) suffered from a recurrent feeling of genital prolapse, and 14 patients (28%) had urinary symptoms postoperatively. CONCLUSIONS: Our treatment showed good anatomical corrections based on chain cystourethrography, and there were no prognostic differences among the surgical procedures. However, anatomical correction does not always mean improvement of urinary symptoms. Further studies are needed to clarify what factors contribute to the prognosis in such cases. PMID- 11049241 TI - Clinical applications of serum placental protein 14 (PP14) measurement in the IVF ET cycle. AB - OBJECTIVE: Placental protein 14 (PP14) is known to be one of the endometrial proteins that reflect endometrial functioning throughout the menstrual cycle. In this study, we examined PP14 as a marker for human endometrial receptivity in order to predict the outcome of in vitro fertilization and the embryo-transfer (IVF-ET) cycle. PATIENTS AND METHODS: The subjects were 72 women who had 96 IVF ET cycles and who were examined at Tokyo Medical University Hospital during the period of January 1998 to June 1998 because of mechanical or unexplained infertility for a duration of at least 2 years. Serum samples were collected from all patients during treatment cycles, and serum PP14 concentrations were measured by a newly established enzyme-linked immunosorbent assay (ELISA). RESULTS: In the pregnant group, serum PP14 concentrations were markedly increased after ET, and a significant difference between the pregnant group and the nonpregnant group was observed 8 days following ET (p < 0.01). PP14 concentrations were higher in patients with endometria that exhibited homogenous patterns and that were more than 7 mm thicker than in other patients, as determined by ultrasound on the day of oocyte collection (p < 0.005). The pregnancy rates of patients with homogeneous patterns were lower than those of patients showing a trilaminar pattern. No pregnancies were observed when serum PP14 concentrations were greater than 6.85 U/l on the day of oocyte collection. CONCLUSION: PP14 might be a useful marker for human endometrial receptivity to predict the outcome of IVF-ET cycles. PMID- 11049242 TI - Isolated pericardial effusion and transient abnormal myelopoiesis in a fetus with Down's syndrome. AB - Isolated pericardial effusion was detected in a fetus at 34 weeks of gestation. A male infant weighing 2,044 g was born by cesarean section because of a non assuring fetal heart rate pattern at 35 weeks of gestation. Transient leukocytosis (36,100/microl) with 49% blast cells was seen in this neonate. The infant's karyotype was 47, XY + 21. The pericardial effusion disappeared after treatment with prednisolone at a dose of 2 mg/kg/day. Hypothyroidism was subsequently found. Thus, the subject patient with Down's syndrome developed isolated pericardial effusion, transient abnormal myelopoiesis (TAM), and hypothyroidism. Because more than 20% of the infants with TAM and Down's syndrome develop acute nonlymphocytic leukemia in early childhood, he is being closely observed. PMID- 11049243 TI - Antenatal use of ambroxol for the prevention of infant respiratory distress syndrome. AB - OBJECTIVE: Our purpose was to evaluate the efficacy and safety of ambroxol for the prenatal prophylaxis of infant respiratory distress syndrome (IRDS). STUDY DESIGN: This was a prospective study with 2 groups of pregnant patients with premature labor or with premature rupture of membranes at an estimated gestation between 27 to 34 completed weeks. Ambroxol treatment group consisted of 39 subjects in whom 1,000 mg of ambroxol diluted in 500 ml of 5% glucose solution was given intravenously for 4 hours once a day for 3 days, while the control group consisted of 41 subjects in whom ambroxol was not administered. Main measures included Apgar scores, clinical signs of one or more of the following: respiratory rate of > 60/min, intercostal retraction, alar flaring, expiratory grunting, cyanosis on room air and radiological evidence of IRDS. Chi-square test was used to determine the statistical significance of the results. RESULTS: Tolerable maternal side effects were noted. Profile of newborns delivered were similar in both groups. Incidence of IRDS was significantly less in the treatment group (p < 0.01). CONCLUSIONS: Antenatal administration of ambroxol resulted in a significant decrease in the incidence of IRDS as well as perinatal morbidity and mortality. Due to the efficacy and safety of this drug, it might be useful for the prevention of IRDS. PMID- 11049663 TI - Behavioral and brain wave evidence for automatic processing of orthographically regular letter strings. AB - Evidence for the automatic processing of nonword letter strings was sought in subjects who performed two different perceptual tasks. Twelve subjects detected valid words. Twelve other subjects detected Gestalt patterns that contained no letters. The targets and two types of nonword letter strings were interspersed in a stream of nonletter images. Behavioral data showed significant effects of orthographic regularity of letter strings in the group that detected words, but not in the group that detected Gestalt patterns. Brain-wave responses showed the effect of orthographic regularity in both groups. The responses were significantly larger for regular than for irregular strings. This showed that brain waves can be used to study the operation of reading processes in subjects who are performing a task that does not require reading ability. PMID- 11049664 TI - Identification reaction times of voiced/voiceless continua: a right-ear advantage for VOT values near the phonetic boundary. AB - We explored the degree to which the duration of acoustic cues contributes to the respective involvement of the two hemispheres in the perception of speech. To this end, we recorded the reaction time needed to identify monaurally presented natural French plosives with varying VOT values. The results show that a right ear advantage is significant only when the phonetic boundary is close to the release burst, i.e., when the identification of the two successive acoustical events (the onset of voicing and the release from closure) needed to perceive a phoneme as voiced or voiceless requires rapid information processing. These results are consistent with the recent hypothesis that the left hemisphere is superior in the processing of rapidly changing acoustical information. PMID- 11049666 TI - Conceptual structure and the structure of concepts: a distributed account of category-specific deficits. AB - We present a new account of the fine-grained structure of semantic categories derived from neuropsychological, behavioral, and developmental data. The account places theoretical emphasis on the functions of the referents of concepts. We claim (i) that the distinctiveness of functional features correlated with perceptual features varies across semantic domains; and (ii) that category structure emerges from the complex interaction of these variables. The representational assumptions that follow from these claims make strong predictions about what types of semantic information are preserved in patients showing category-specific deficits following brain damage. These claims are illustrated with a connectionist simulation which, when damaged, shows patterns of preservation of distinctive and shared functional and perceptual information which varies across semantic domains. The data model both dissociations between knowledge for artifacts and for living things and recent neuropsychological evidence concerning the robustness of functional information in the representation of concepts. PMID- 11049665 TI - Is overt stuttered speech a prerequisite for the neural activations associated with chronic developmental stuttering? AB - Four adult right-handed chronic stutterers and four age-matched controls completed H(2)(15)O PET scans involving overt and imagined oral reading tasks. During overt stuttered speech prominent activations occurred in SMA (medial), BA 46 (right), anterior insula (bilateral), and cerebellum (bilateral) plus deactivations in right A2 (BA 21/22). These activations and deactivations also occurred when the same stutterers imagined they were stuttering. Some parietal regions were significantly activated during imagined stuttering, but not during overt stuttering. Most regional activations changed in the same direction when overt stuttering ceased (during chorus reading) and when subjects imagined that they were not stuttering (also during chorus reading). Controls displayed fewer similarities between regional activations and deactivations during actual and imagined oral reading. Thus overt stuttering appears not to be a prerequisite for the prominent regional activations and deactivations associated with stuttering. PMID- 11049667 TI - Sentence repetition and processing resources in Alzheimer's disease. AB - Sentence processing in Alzheimer's disease (AD) has been found to be influenced by several grammatical and extragrammatical factors, including phrase structure and verb-argument relations, number of propositions/verbs, and processing resource capacity. This study examines the effects of these variables on sentence production in AD. Normal control and AD subjects were asked to repeat six types of sentences varying along the above dimensions of complexity. Subjects' processing resource capacity was measured using several verbal working memory tests. AD subjects' sentence-repetition performance was impaired compared to the normal control group. Significant effects were observed for branching direction of phrase structure, canonicity of verb-argument relations, and serial position of errors. Sentence-repetition performance significantly correlated with working memory scores. The findings are interpreted within a resource capacity theory of sentence processing. PMID- 11049668 TI - Articulatory/phonetic sequencing at the level of the anterior perisylvian cortex: a functional magnetic resonance imaging (fMRI) study. AB - Damage to the anterior peri-intrasylvian cortex of the dominant hemisphere may give rise to a fairly consistent syndrome of articulatory deficits in the absence of relevant paresis of orofacial or laryngeal muscles (apraxia of speech, aphemia, or phonetic disintegration). The available clinical data are ambiguous with respect to the relevant lesion site, indicating either dysfunction of the premotor aspect of the lower precentral gyrus or the anterior insula in the depth of the Sylvian fissure. In order to further specify the functional anatomic substratum of this syndrome, functional magnetic resonance imaging (fMRI) was performed during reiteration of syllables differing in their demands on articulatory/phonetic sequencing (CV versus CCCV versus CVCVCV). Horizontal tongue movements and a polysyllabic lexical item served as control conditions. Repetition of the CV and CCCV monosyllables elicited a rather bilateral symmetric hemodynamic response at the level of the anterior and posterior bank of the central sulcus (primary sensorimotor cortex), whereas a more limited area of neural activity arose within this domain during production of lexical and nonlexical polysyllables, significantly or exclusively lateralized toward the left hemisphere. There is neurophysiological evidence that primary sensorimotor cortex mediates the "fractionation" of movements. Assuming that the polysyllables considered are organized as coarticulated higher-order units, the observed restricted and lateralized cortical activation pattern, most presumably, reflects a mode of "nonindividualized" motor control posing fewer demands on "movement fractionation." These findings may explain the clinical observation of disproportionately worse repetition of trisyllabic items as compared to monosyllables in apraxia of speech. The various test materials failed to elicit significant activation of the anterior insula. If at all, only horizontal tongue movements yielded a hemodynamic reaction extending beyond the sensorimotor cortex to premotor areas. Since limbic projections target the inferior dorsolateral frontal lobe, the enlarged region of activation during horizontal tongue movements might reflect increased attentional requirements of this task. PMID- 11049669 TI - Phonological and articulatory impairment in Alzheimer's disease: a case series. AB - We demonstrate that phonological and articulatory impairments may occur at presentation or early in the course of Alzheimer's disease, contrary to claims that these aspects of language production are relatively preserved until the final stages of this disease. Six patients with pathologically confirmed Alzheimer's disease (AD) and four patients with clinically diagnosed dementia of the Alzheimer's type (DAT) presented with one of five different clinical profiles: nonfluent progressive aphasia, mixed progressive aphasia, progressive aphasia diagnosed as DAT from neuropsychological assessment, initial amnestic syndrome with prominent phonological errors, and biparietal syndrome. Analysis of their conversational speech, single-word production, and performance of highly familiar series speech tasks such as counting revealed false start errors, phonological paraphasias, and/or articulatory difficulty. Neuropathological changes were located in left perisylvian regions consistent with speech and language impairment but atypical for Alzheimer's disease. PMID- 11049670 TI - The Crystallization of Hydroxyapatite in the Presence of Lysine. AB - Lysine, which is an amino acid with a basic side group, is present in biological fluids and its role in the biological calcification process was investigated. It was found to inhibit the crystal growth rate of hydroxyapatite (HAP), in solutions supersaturated only with respect to this calcium phosphate salt and this rate reduction was attributed to adsorption and further blocking of the active growth sites on the crystal surface. The crystallization kinetics were interpreted in terms of the Langmuir adsorption model. The apparent order of the crystallization reaction was found to be n=2, suggesting a surface diffusion controlled spiral growth mechanism. Kinetic results of HAP crystallization were obtained using the constant composition method where the concentration of the reactants is kept constant during the course of the crystal growth experiment. Copyright 2000 Academic Press. PMID- 11049671 TI - Ancient Parchment Examination by Surface Investigation Methods. AB - A restoring process was set up to restore flexibility, size, and shape in naturally aged or fire-damaged parchments of old manuscripts. Validation of such a process requires the measurement of intrinsic parchment properties and comparison of them before and after the treatment. To this aim, we investigated morphological, mechanical, and surface physico-chemical properties of parchment by taking SEM pictures and characterizing small samples by microindentation, mercury porosimetry, and water vapor adsorption/desorption isotherms. Copyright 2000 Academic Press. PMID- 11049672 TI - Influence of Temperature on Stability of Electrostatically Stabilized Alumina Suspensions. AB - Pure electrostatically stabilized aqueous alumina suspensions were prepared at various solid loadings in order to study the influence of temperature on the surface charge properties and rheology. Surface charge density at various temperatures was measured through potentiometric titration, and the analysis of the potentiometric data was accomplished using the constant capacitance surface model. Calculations of the pair-wise interaction potential between charged colloidal spheres dispersed in water were then carried out using conventional DLVO theory and a software package Stabil45, taking into account the temperature dependence of surface charge density and dielectric constant. The results showed that increasing temperature leads to a gradual diminution of alumina surface ionization, dielectric constant, and a total energy barrier for coagulation. The enhancement of the coagulation rate with increasing temperature leads to a gradual increase of both relative apparent viscosity and thixotropy. Copyright 2000 Academic Press. PMID- 11049673 TI - An Experimental Study of the Kinetics of Particle Deposition in a Wall-Jet Cell Using Total Internal Reflection Microscopy. AB - The deposition of polystyrene latex particles of 0.46 um diameter was studied in situ using a wall-jet cell in combination with total internal reflection microscopy. The particles were deposited onto an indium tin oxide surface in a laminar flow field for up to 13 h. The initial particle flux was found to be mass transfer-controlled. It was shown that one period of the time-dependent deposition rate period was of first-order nature. The effective particle transfer coefficient during this period appeared to be correlated inversely to the wall shear rate. With the help of three characteristic empirical constants, one of them being a mass transfer coefficient, the overall deposition process was described by a model equation. The concentration dependency was elucidated using a Langmuir-type pseudoisotherm. Copyright 2000 Academic Press. PMID- 11049674 TI - Stokesian Dynamics Simulations of Ferromagnetic Colloidal Dispersions Subjected to a Sinusoidal Shear Flow. AB - We have conducted Stokesian dynamics simulations to investigate the dynamic properties of ferromagnetic colloidal dispersions subjected to a sinusoidal shear flow. Thick chain-like cluster formation is significantly influenced by an oscillatory shear flow even if the amplitude is relatively small, since the internal structures of thick chain-like clusters are highly sensitive to the change in the direction of the shear flow. The motion of thick chain-like clusters is out of phase to a sinusoidal shear rate, and the phase difference is strongly correlated with that of the viscosity and normal stress coefficients. The viscoelastic properties become more apparent with decreasing frequency of the oscillatory shear flow, since such properties have a strong relationship with the thick chain-like cluster formation. In other words, since thick chain-like clusters are more stable for the case of a smaller frequency shear flow, such stable clusters induce significant viscoelastic properties of ferromagnetic colloidal dispersions in a strong, applied magnetic field. Copyright 2000 Academic Press. PMID- 11049675 TI - Physical Properties of Magnetic Fluids and Nanoparticles from Magnetic and Magneto-rheological Measurements. AB - Static magnetization curves and the magnetorheological effect were used to study the microstructural properties (agglomerate formation) of magnetic fluids and the properties of dispersed nanoparticles. Improved techniques for magnetogranulometry analysis and a formula for the magnetoviscous effect were proposed. The area of applicability of some existing models was studied. The density, distribution, and dimension of particles, as well as the thickness of the nonmagnetic layer were accurately determined from magnetic measurements. The Shliomis diameter and the effective anisotropy constant were determined from rheological and magnetorheological measurements using information obtained from magnetization curves. Copyright 2000 Academic Press. PMID- 11049676 TI - Synthesis and Surface-Active Properties of New Photosensitive Surfactants Containing the Azobenzene Group. AB - Several water-soluble cationic surfactants, 4-alkylazobenzene-4'-(oxy-2 hydroxypropyl)trimethylammonium methylsulfate (AZMS) (AZMS-0, AZMS-1, AZMS-2, AZMS-4, and AZMS-8), containing alkylglycidylether and azoarene have been synthesized with high yields of 63-78% and their surface-active properties have been investigated upon irradiation with UV/vis light. All of the trans-AZMS surfactants are isomerized to cis-trans mixtures containing 92.5% cis isomer by UV light irradiation at 350 nm. The cis isomers in the mixtures are reverted to trans isomers by visible light irradiation (lambda>445 nm). Such photoisomerization induces changes in the surface activity of each surfactant. The critical micelle concentration (cmc) of the trans form of AZMS-8 surfactant is about 1.28x10(-4) mol/l. At the photostationary state, 92.5% of the trans form is changed to the cis form which exhibits a slightly higher cmc (3.41x10(-4) mol/l). The new cmc of AZMS surfactants upon photoisomerization is similar to that of the ideal mixed micellar system. In particular, the ratio of cmc(cis) to cmc(trans) of AZMS derivatives is about 1.87-2.85 which increases proportionally with the chain length of alkyl group. The minimum average area per molecule (A(min)(a/w)) for the trans and cis isomers of AZMS-8 is 0.60 and 0.74 nm(2), respectively. The difference in the A(min)(a/w) may originate from the structural differences in the two isomers. These values are quite different as compared to those of the conventional azobenzene surfactants. Copyright 2000 Academic Press. PMID- 11049677 TI - Particle Interactions in Diffusiophoresis and Electrophoresis of Colloidal Spheres with Thin but Polarized Double Layers. AB - The diffusiophoretic and electrophoretic motions of two colloidal spheres in the solution of a symmetrically charged electrolyte are analyzed using a method of reflections. The particles are oriented arbitrarily with respect to the electrolyte gradient or the electric field, and they are allowed to differ in radius and in zeta potential. The thickness of the electric double layers surrounding the particles is assumed to be small relative to the radius of each particle and to the gap width between the particles, but the effect of polarization of the mobile ions in the diffuse layer is taken into account. A slip velocity of fluid and normal fluxes of solute ions at the outer edge of the thin double layer are used as the boundary conditions for the fluid phase outside the double layers. The method of reflections is based on an analysis of the electrochemical potential and fluid velocity disturbances produced by a single dielectric sphere placed in an arbitrarily varying electrolyte gradient or electric field. The solution for two-sphere interactions is obtained in expansion form correct to O(r(12)(-7)), where r(12) is the distance between the particle centers. Our analytical results are found to be in good agreement with the available numerical solutions obtained using a boundary collocation method. On the basis of a model of statistical mechanics, the results of two-sphere interactions are used to analytically determine the first-order effect of the volume fraction of particles of each type on the mean diffusiophoretic and eletrophoretic velocities in a bounded suspension. For a suspension of identical spheres, the mean diffusiophoretic velocity can be decreased or increased as the volume fraction of the particles is increased, while the mean electrophoretic velocity is reduced with the increase in the particle concentration. Generally speaking, the particle interaction effects can be quite significant in typical situations. Copyright 2000 Academic Press. PMID- 11049678 TI - Adsorption of Immunoglobulin G on Core-Shell Latex Particles Precoated with Chaps. AB - The aim of this work is to investigate the adsorption behavior of a monoclonal antibody (immunoglobulin G, IgG) on latex particles, possessing reactive chloromethyl groups, precoated with 3-([3-cholamidopropyl]dimethylammonio-1 propanesulfonate (Chaps). The amount and reactivity of the surface chloromethyl groups were monitored by the nucleophilic attack of glycinate to the functional groups as a function of time at 22 and 36 degrees C. The extent of displacement of Chaps by IgG and the enthalpy of the process were determined under two different conditions of precoating the latex particles with Chaps, at 22 and 36 degrees C. The adsorption of IgG takes place in two steps; the first one involves physical interaction between IgG and the surface. This step is relatively fast (in the range of minutes) and independent of temperature. In the second step covalent bonding between the protein and the active surface groups occurs. This reaction is improved by raising the temperature because Chaps desorption, which exposes the reactive chloromethyl groups on the latex particles, is kinetically and thermodynamically favored at 36 degrees C and the covalent bonding of IgG is faster at 36 degrees C. Copyright 2000 Academic Press. PMID- 11049679 TI - The Effect of Alcohol Solvents on the Porosity and Phase Composition of Titania. AB - Bimodally porous titania powders were made by hydrolysis of titanium tetraisopropoxide (TTIP) dissolved in various alcohols (methanol, ethanol, isopropanol, and sec-butanol). The specific surface area (SSA) of the powders dried at 150 degrees C ranged from 332 to 624 m(2)/g as determined by nitrogen adsorption. At excess alcohol concentration, the SSA of the dried powders decreased in the order of sec-butanol, iso-propanol, ethanol, and methanol at a constant alcohol/TTIP molar ratio. The pore size distribution was bimodal with fine intraparticle pore diameters at 1-6 nm and larger interparticle pore diameters at 30-120 nm as determined by nitrogen adsorption isotherms. The average intraparticle pore diameter decreased with increasing alcohol concentration for methanol and ethanol, while it was rather constant at 3.3 nm, irrespective of alcohol concentration for iso-propanol and sec-butanol. The evolution of particle phase composition was determined by X-ray diffraction ranging from amorphous to crystalline anatase and rutile largely proportional to the calcination temperature and to a lesser extent on the type and concentration of alcohols. Copyright 2000 Academic Press. PMID- 11049680 TI - Effect of Deposition in Deep-Bed Filtration: Determination and Search of Rate Parameters. AB - Particle deposition in deep-bed filters (porous media) is, by nature, an unsteady state process and the extent of deposition plays an important role in determining filter performance. A general method of establishing the relationship between deposition rate and the extent of deposition from effluent concentration history of filters is presented. The efficacy of the method is examined and its utility demonstrated through its applications to certain experimental data. Copyright 2000 Academic Press. PMID- 11049681 TI - Comparison of Different Approaches to the Determination of the Work of Critical Cluster Formation. AB - A comparative analysis of the results of determination of the work of critical cluster formation in nucleation theory for three different methods of evaluation Gibbs' method, the van der Waals-Cahn and Hilliard, and a newly developed modified Gibbs' approach-is given. As a model system for comparison, regular solutions are chosen. In addition to the work of critical cluster formation, the composition of the critical clusters, their characteristic sizes and the values of the surface tension are determined in dependence on the initial supersaturation in the system or, equivalently, on the size of the critical clusters. It is found, in particular, that, for regular solutions, Tolman's equation cannot serve as a first approximation for the description of the curvature dependence of the surface tension even for large cluster sizes and an alternative formula is developed. It is shown that the latter two mentioned methods of determination of the work of critical cluster formation (the van der Waals-Cahn and Hilliard and the modified Gibbs' approach) lead-at least for the model system considered-to qualitatively and partly quantitatively equivalent results. Nevertheless, differences remain which may lead to quantitative deviations when applied to the determination of the steady-state nucleation rates. The possible origin of such deviations is discussed and some further directions of analysis are anticipated. Copyright 2000 Academic Press. PMID- 11049682 TI - Kinetics of Adsorption and Desorption of Poly(ethylenimine) and Its Complex with Copper(II) on Copper(II) Oxide Powder. AB - Adsorption of poly(ethylenimine) and complexes of poly(ethylenimine) with copper(II) ions on copper(II) oxide powder has been investigated. The rate of adsorption and reaction between poly(ethylenimine) and CuO is very fast. The desorption rate of the complex of poly(ethylenimine) with copper(II) ions is slower and decreases when the initial polymer concentration is decreased. The desorption rate passes through a minimum when the initial concentration of CuO powder in the mixture increases. A mathematical model of polymer adsorption and desorption is proposed. Copyright 2000 Academic Press. PMID- 11049683 TI - Sorption Studies of Cobalt(II) on Colloidal Hematite Using Potentiometry and Radioactive Tracer Technique. AB - The sorption of Co(II) on colloidal hematite was studied as a function of pH, ionic strength, and Co(II) concentration. Two different techniques were used, yielding two different sets of information: (i) potentiometric titrations that provide information on the number of protons released as a function of pH owing to the sorption of Co(II) and (ii) measurement of the amount of cobalt sorbed on the surface as a function of pH using a radioactive tracer, (60)Co. At low Co(II) concentrations (10(-8) M), the sorption was found to be independent of ionic strength but there seems to be a weak ionic strength dependence at higher Co(II) concentrations (10(-4) M). The adsorption edge moved to higher pH with increasing Co(II) concentration. For the high Co(II) concentration, the number of protons released per cobalt sorbed increased from zero to approximately 1.5. The basic charging properties of hematite were modeled with four different surface complexation models. The 1-pK Basic Stern Model (BSM), with binding of electrolyte ions to the Stern plane, seems to be the most reasonable model if the ambition is to describe experimental data at different ionic strengths. The sorption of cobalt was modeled with the 1-pK BSM. By introducing a low concentration of high affinity surface sites for cobalt sorption it was possible to model the sorption in very wide cobalt concentrations, ranging from 10(-8) M to 10(-4) M. Copyright 2000 Academic Press. PMID- 11049684 TI - Local Structure Evolution in Particle Network Formation Studied by Brownian Dynamics Simulation. AB - The effect of solid content and colloidal interactions on the structure of forming networks of colloidal particles is studied by Brownian dynamics simulation. The different situations are compared in terms of the pair distribution function and the distribution of nearest neighbors around each particle. The results indicate that, in fast coagulation, the higher solid contents lead to a freezing-in of the liquid structure. Nevertheless, this effect can be reduced substantially by the introduction of a shallow secondary minimum and an energy barrier in the interaction potential. However, the structures resulting from such slow coagulation show a substantial degree of porosity, larger than those produced at the same solid content but by fast coagulation. It is also shown how the porosity (defined on a few particle diameters) is reflected in the distribution of nearest neighbors around the center particle, i.e., the very local conformation in the particle network. Fractal analysis shows that, at the relatively high volume fractions considered in this study, no intermediate fractal regime exists. Copyright 2000 Academic Press. PMID- 11049685 TI - The Dielectric Function for Water and Its Application to van der Waals Forces. AB - The dielectric response, varepsilon(ixi), for water (which is required in Lifshitz theory to calculate the van der Waals interactions in aqueous systems) is commonly constructed, in the absence of complete spectral data, by fitting a damped-harmonic-oscillator model to absorption data. Two sets of parameters for the model have been developed corresponding to different constraints: Parsegian and Weiss (J. Colloid Interface Sci., 1981, 81, 285) and Roth and Lenhoff (J. Colloid Interface Sci., 1996, 179, 637). These different representations of the dielectric response lead to significant differences in the van der Waals force calculated from Lifshitz theory. In this work, more recent and complete spectral data for water were compiled from the literature and direct integration of the Kramers-Kronig relations was used to construct a new varepsilon(ixi) for water at 298 degrees K. This approach also allows a number of different types of spectral measurements (such as infrared spectroscopy, microwave resonance techniques, and x-ray inelastic scattering) in the compilation of absorption data over a large frequency range (on the order of 8 to 10 decades in frequency). A Kramers-Kronig integration was employed to construct the real and imaginary parts of varepsilon(omega), varepsilon'(omega), and varepsilon"(omega) for water from the different spectral measurements before calculation of varepsilon(ixi) from its integral definition. The resulting new varepsilon(ixi) is intermediate between the Parsegian-Weiss and Roth-Lenhoff representations of varepsilon(ixi), does not use a model, and treats the conversion of absorption data as rigorously as possible. We believe the varepsilon(ixi) from the present work is the most reliable construction for use in van der Waals force calculations using Lifshitz theory. The extension of the varepsilon(ixi) construction to other temperatures is also discussed. Copyright 2000 Academic Press. PMID- 11049686 TI - Adsorption and Precipitation of Aqueous Zn(II) on Alumina Powders. AB - The products of aqueous Zn(II) sorption on high-surface-area alumina powders (Linde-A) have been studied using XAFS spectroscopy as a function of Zn(II) sorption density (Gamma=0.2 to 3.3 umol/m(2)) at pH values of 7.0 to 8.2. Over equilibration times of 15-111 h, we find that at low sorption densities (Gamma=0.2-1.1 umol/m(2)) Zn(II) forms predominantly inner-sphere bidentate surface complexes with AlO(6) polyhedra, whereas at higher sorption densities (Gamma=1.5 to 3.5 umol/m(2)), we find evidence for the formation of a mixed-metal Zn(II)-Al(III) hydroxide coprecipitate with a hydrotalcite-type local structure. These conclusions are based on an analysis of first- and second-neighbor interatomic distances derived from EXAFS spectra collected under ambient conditions on wet samples. At low sorption densities the sorption mechanism involves a transformation from six-coordinated Zn-hexaaquo solution complexes (with an average Zn-O distance of 2.07 A) to four-coordinated surface complexes (with an average Zn-O distance of 1.97 A) as described by the reaction identical withAl(OH(a))(OH(b))+Zn (H(2)O)(6)(2+)--> identical withAl(OH(a)') (OH(b)')Zn(OH(c)')(OH(d)'+4H(2)O+zH(+), where identical withAl(OH(a))(OH(b)) represents edge-sharing sites of Al(O,OH,OH(2))(6) octahedra to which Zn(O,OH,OH(2))(4) bonds in a bidentate fashion. The proton release consistent with this reaction (z=a-a'+b-b'+4-c'-d'), and with bond valence analysis falls in the range of 0 to 2 H(+)/Zn(II) when hydrolysis of the adsorbed Zn(II) complex is neglected. This interpretation suggests that proton release is likely a strong function of the coordination chemistry of the surface hydroxyl groups. At higher sorption densities (1.5 to 3.5 umol/m(2)), a high-amplitude, second-shell feature in the Fourier transform of the EXAFS spectra indicates the formation of a three dimensional mixed-metal coprecipitate, with a hydrotalcite-like local structure. Nitrate anions presumably satisfy the positive layer charge of the Al(III)-Zn(II) hydroxide layers in which the Zn/Al ratio falls in the range of 1 : 1 to 2 : 1. Our results for the higher Gamma-value sorption samples suggest that Zn hydrotalcite-like phases may be a significant sink for Zn(II) in natural or catalytic systems containing soluble alumina compounds. Copyright 2000 Academic Press. PMID- 11049687 TI - Application of the Radiotracer Technique for the Study of the Specific Anion Adsorption on Al(2)O(3) in Acidic Medium. AB - The specific adsorption of radiolabeled anions (sulfate, chloride) from perchlorate supporting electrolyte on gamma-Al(2)O(3) powder has been investigated. It was demonstrated that a measurable adsorption occurs only at pH values under pH 6. The adsorption capacity of a given amount of gamma-Al(2)O(3) powder tends to a limiting value with decreasing pH. A Langmuir-like adsorption isotherm describes the concentration dependence of the adsorption of sulfate species. It was found that the experimental results are very similar to those obtained in the case of hematite. Copyright 2000 Academic Press. PMID- 11049688 TI - A Nonlinear Rupture Theory of Thin Liquid Films with Soluble Surfactant. AB - The effects of soluble surfactant on the dynamic rupture of thin liquid films are investigated. A nonlinear coupling evolution equation is used to simulate the motion of thin liquid films on free surfaces. A generalized Frumkin model is adopted to simulate the adsorption/desorption kinetics of the soluble surfactant between the surface and the bulk phases. Numerical simulation results show that the liquid film system with soluble surfactant is more unstable than that with insoluble surfactant. Moreover, a generalized Frumkin model is substituted for the Langmuir model to predict the instability of liquid film with soluble surfactant. A numerical calculation using the generalized Frumkin model shows that the surfactant solubility increases as the values of parameters of absorption/desorption rate constant (J), activation energy desorption (nu(d)), and bulk diffusion constant (D(1)) increase, which consequently causes the film system to become unstable. The surfactant solubility decreases as the rate of equilibrium (lambda) and interaction among molecules (K) are increased, which therefore stabilizes the film system. On the other hand, an increase of relative surface concentration (the index of a power law), beta(n), will initially result in a decrease of corresponding shear drag force as beta and n increase from 0 to 0.3 and 0.85, respectively. This will enhance the Marangoni effect. However, a further increase of beta and n to greater than 0.3 and 0.85, respectively, will conversely result in an increase of the corresponding shear drag force. This will weaken the Marangoni effect and thus result in a reduction of interfacial stability. Copyright 2000 Academic Press. PMID- 11049689 TI - Hamaker Constants of Systems Involving Water Obtained from a Dielectric Function That Fulfills the f Sum Rule. AB - Hamaker constants of systems involving liquid water are evaluated, within the full Lifshitz theory, by means of a recently proposed model of the dielectric function of this substance [Dingfelder et al., Radiat. Phys. Chem. 53, 1 (1998)], which has been extended in the present work by including terms corresponding to infrared excitations and microwave relaxation. An important feature of the complete model is that, besides a good fit to experimental data, it satisfies the physical constraint provided by the f sum rule. For symmetrical systems interacting across water, calculated Hamaker constants are generally in good agreement with results obtained using the Ninham-Parsegian representation with the Roth and Lenhoff parameters for water. Copyright 2000 Academic Press. PMID- 11049690 TI - Dissolutive Capillary Penetration with Expanding Pores and Transient Contact Angles. AB - The penetration kinetics of a cylindrical capillary and a capillary porous body with a temporally expanding capillary radius due to reactive dissolution ahead of the liquid front is modeled under conditions where the equilibrium contact angle is not attained during at least part of the penetration process. These effects cause deviations from the predictions of the Washburn equation, with the actual penetration kinetics depending upon the rate processes involved. Copyright 2000 Academic Press. PMID- 11049691 TI - Synthesis of Monodisperse Al-Substituted Hematite Particles from Highly Condensed Metal Hydroxide Gels. AB - Monodispersed aluminum-substituted hematite particles were synthesized from concentrated mixed solutions of FeCl(3) and AlCl(3) using the gel/sol process. With increasing aluminum content (Al/(Fe+Al)) in the starting solutions, hematite particles became more and more elongated. The product particles exhibited ellipsoidal, rod- and peanut-like shapes, depending on Al concentration. Higher Al content (>20 mol%) in the system greatly suppressed the formation of hematite. Atomic emission spectrometry analysis and lattice parameter measurement confirmed the substitution of Al in hematite particles. Copyright 2000 Academic Press. PMID- 11049692 TI - Why is glycated LDL more sensitive to oxidation than native LDL? A comparative study. AB - It is well established that oxidative modification of low-density lipoprotein (LDL) plays a causal role in human atherogenesis and the risk of atherosclerosis is increased in patients with diabetes mellitus. To examine the influence of different agents which may influence LDL-glycation and oxidation, experiments including glycation with glucose, glucose 6-phosphate, metal chelators (EDTA) and antioxidants (BHT) were performed. The influence of time dependence on the glycation process and the alteration of the electrophoretic mobility of LDL under diverse glycation and/or oxidation conditions was also investigated. The formation of conjugated dienes and levels of lipid peroxides in these different LDL-modifications were estimated. The copper-induced oxidation of LDL in vitro was determined by measurement of thiobarbituric acid reactive substances (TBARS) and expressed as nmol MDA/mg of LDL protein. We found that glycated LDL is more prone to oxidation than native LDL. Using native LDL, the maximal oxidation effect was found to reach a value of 49.72 nmol MDA/mg protein after 8 h. The maximum oxidation of the 31 days, glycated LDL with glucose was 71.76 nmol MDA/mg protein amounting to 144.33% of the value found for native LDL. In the case of glucose 6-phosphate glycation, the maximum oxidation under the same conditions amounted to 173.77% of the value found for native LDL. To measure the extent of glycation, fluorescence of advanced glycation end products (AGEs) was determined (370 nm excitation and 440 nm emission). The most potent glycation agent was glucose 6-phosphate leading to the formation of very high amounts of AGEs. This process was promoted in the absence of EDTA, which prevents the oxidative cleavage of modified Amadori products (ketoamines) to AGEs. We therefore conclude that both processes, glycation and oxidation, result in the modification of LDL. The lower the glycation-rate (+/- EDTA) as measured by relative fluorescence units RFU (generation of AGEs), the lower the additional oxidation rate after glycation as measured by TBARS (generation of MDA equivalents). Glycation and/or oxidation change the electrophoretic mobility of LDL. PMID- 11049693 TI - A direct method for the simultaneous measurement of ceramide and phospholipase D activity. AB - Both ceramide and phospholipase D (PLD) have important roles in a variety of signal transduction pathways. Recent evidence suggests that ceramide is a novel second messenger with specific biological effects. Publications in this field have increased rapidly in the last few years. However, a method to directly and rapidly measure cermide production has been lacking. Herein, we report on a novel, inexpensive, direct and rapid assay for the measurement of ceramide and the simultaneous measurement of PLD activity. This method uses labeling of cells with [(14)C]myristic acid and a TLC solvent of ethyl acetate/acetic acid/trimethylpentane. This method avoids the loss of radioactivity and variability due to changes in DAG kinase activity that are associated with the commonly-used DAG kinase assay. PMID- 11049694 TI - Nutritional status and intermediate chain-length fatty acids influence the conservation of essential fatty acids in the milk of northern Nigerian women. AB - The milk of 89 women in northern Nigeria was analyzed for the fatty acid composition of the total milk lipids, and assessed for the effect nutritional status has on the conservation of essential and non-essential fatty acids when the proportions of C(10)-C(14)fatty acids are increased. The women were stratified on the basis of their body mass index, and calculations were made to estimate the effects of a 3.3-fold increase in the proportion of C(10)-C(14)fatty acids on the proportion of alpha-linolenic acid, docosahexaenoic acid, linoleic acid and arachidonic in total milk lipids. In the well-nourished group (group III, body mass index >23 kg/m(2)), the critical n-3 and n-6 fatty acids were not conserved, while in poorly nourished women (group I, body mass index <19 kg/m(2)), marked conservation of alpha-linolenic acid, docosahexaenoic acid, arachidonic acid, and palmitic acid was seen. Poor nutritional status of the mother appears to promote selective retention of critical essential and non essential fatty acids in the milk lipid fraction. PMID- 11049695 TI - Correlation between the content of intermediate chain-length fatty acids and copper in the milk of Fulani women. AB - Intermediate chain-length fatty acids (C10-C14) in human milk triglycerides provide an easily absorbable fuel that provide a significant amount of the energy needed for growth during the first few months of life. The C10-C14 fatty acid and trace mineral content of human milk is variable. In this report we examined the relationship between the content of calcium, copper, magnesium, manganese, phosphorus, and zinc, and the weight percentage of C10-C14 fatty acids in milk from 33 Fulani women in northern Nigeria between 2 and 24 weeks post-gestation. The milk from these women contained proportions of C10-C14 fatty acids that were comparable to those reported for other populations around the world, as were the concentrations of Ca, Cu, Mg, Mn, Zn and P. Significant correlations were observed between the milk content of Cu and the wt% of C10 (P=0. 005, r=0.475), C12 (P=0.001, r=0.539), C14 (P=0.44, r=0.352) and the total intermediate chain length fatty acids (P=0.008, r=0.450). No correlations were observed between these fatty acids and any of the other five minerals. We speculate that the relationship between Cu and fatty acids could be related to a requirement for Cu by an enzyme required for C10-C14 fatty acid biosynthesis (e.g. decanoyl deacylase) in mammary tissue, or to some unique Cu binding properties of the intermediate chain length fatty acids. PMID- 11049696 TI - Urinary prostaglandin excretion in pregnancy: the effect of dietary sodium restriction. AB - INTRODUCTION: Dietary sodium restriction results in activation of the renin angiotensin-aldosterone-system. In the non-pregnant situation renin release in response to a low sodium diet is mediated by prostaglandins. We studied the effect of dietary sodium restriction on urinary prostaglandin metabolism in pregnancy. PATIENTS AND METHODS: In a randomized, longitudinal study the excretion of urinary metabolites of prostacyclin (6-keto-PGF(1 alpha)and 2,3 dinor-6-keto-PGF(1 alpha)) and thromboxane A(2)(TxB(2)and 2,3-dinor-TxB(2)) was determined throughout pregnancy and post partum in 12 women on a low sodium diet and in 12 controls. RESULTS: In pregnancy the excretion of all urinary prostaglandins is increased. The 6-keto-PGF(1 alpha)/ TxB(2)-ratio as well as the 2, 3-dinor-6-keto-PGF(1 alpha)/ 2,3-dinor-TxB(2)-ratio did not significantly change in pregnancy. CONCLUISION Prostacyclin and thromboxane do not seem to play an important role in sodium balance during pregnancy. PMID- 11049697 TI - Thromboxane-B(2) levels in serum of rabbits receiving a single intravenous dose of aqueous extract of garlic and onion. AB - We have shown previously that fresh garlic extract is effective in reducing thromboxane formation by platelets both in vivo and in vitro animal models of thrombosis. In the present study, the effect of different concentrations of a single dose of aqueous extracts of garlic and onion were evaluated on serum thromboxane-B(2)synthesis in rabbits. Different concentrations of garlic and onion were administered as single doses in the ear vein of rabbits. Rabbits were bled before and at different intervals after the infusion of garlic or onion extracts. Venous blood was collected and allowed to clot at 37 degrees C for 1 h. Thromboxane-B(2)level was measured in the serum by radioimmunoassay. It was observed that garlic inhibits the thrombin-induced platelet synthesis of TXB(2)in a dose-and time-dependent manner. Maximum inhibition of TXB(2)occurred between 0.5 h and 6 h at 25 and 100 mg kg(-1)garlic. At 24 h post-garlic infusion TXB(2)inhibition was reduced to 15% of the control and TXB(2)levels were comparable to that of the control values at 72 h pots-garlic infusion. Infusion of 100 mg kg(-1)onion extract did not elicit any inhibitory effect on TXB(2)synthesis in the serum of rabbit during the treatment period. The rapid recovery of platelet cyclooxygenase activity after infusion of a single dose of garlic suggests that garlic should be taken more frequently in order to achieve beneficial effects in the prevention of thrombosis. PMID- 11049698 TI - Prostanoid receptors in intestinal epithelium: selective expression, function, and change with inflammation. AB - The tissue concentration of PGE(2)is heightened during mucosal inflammation. Nevertheless, the cellular targets of this prostanoid and its effects on epithelial cell physiology are incompletely understood. We used a panel of specific immunoglobulin and mRNA probes in order to localize and quantitate the four member EP family of prostanoid receptors for binding PGE(2)on cells of histologically normal and inflamed human colonic mucosa, and then examined the physiological consequences for the epithelial component of intestine, with special attention to its barrier function. Prostanoid receptors were selectively expressed on a limited number of human colonic mucosal cells, and differed markedly between normal and inflamed tissue. In non-inflamed mucosa, EP(2)and EP(3)were expressed on epithelia at the apex of crypts; while EP(4)was expressed on surface and lateral crypt epithelia. Dual immunostaining and in situ hybridization with digoxygenin-labelled RNA probes largely confirmed the epithelial localization of EP(4). On the other hand, during inflammation, lateral crypt (non-surface) epithelial cells newly and significantly expressed prostanoid receptors EP(2)and EP(3)(p<0.05, by computer-assisted densitometry). Functionally, exogenous E series prostanoids applied to epithelial monolayers in nM concentrations brought about a 24% increase in the level of barrier function; an associated rise in intracellular cAMP (EC(50)of 281); and protection of epithelium from the effects of T cell cytokines. A major perturbation in the number and distribution of functional eicosonoid receptors on epithelia occurs in chronic inflammation of human colonic mucosa. PMID- 11049699 TI - Nosocomial infective endocarditis. AB - SUMMARY: Nosocomial infective endocarditis (NIE) is a rare complication of nosocomial bacteraemia; however, it is an infection of great importance because of its high mortality and because in many cases it is potentially preventable. Whilst many aspects of NIE are similar to community-acquired infective endocarditis (CIE), there are important differences between the two, most notably the predisposing factors, microbial aetiology and prognosis. The diagnosis of NIE is often difficult as many patients have severe underlying disease and coexistent infection elsewhere. Many of these infections could potentially be prevented by the identification of high risk patients, careful assessment of positive blood cultures and effective treatment of bacteraemia in high risk patients. The use of prophylactic antimicrobials in the prevention of infective endocarditis is unproven, however, it is recommended that prophylaxis be considered for certain invasive hospital-based procedures. PMID- 11049700 TI - Problems and solutions in hospital-acquired bacteraemia. AB - Despite infection control efforts, bacteraemia remains one of the most frequent and challenging hospital-acquired infections and is associated with high attributable morbidity and mortality and additional use of healthcare resources. Prevention and control of hospital-acquired blood-stream infection requires improved detection methods, better definition of patient populations at risk, more refined guidelines for the interpretation of positive blood cultures and a better discrimination between sporadic contaminants and true bacteraemia. These issues are addressed in the current review together with those related to the diagnosis, management and recent advances in the prevention of cathether-related bacteraemia, the leading cause of hospital-acquired blood-stream infection. Finally, the reasons and perspectives for blood-stream infection surveillance are briefly discussed. PMID- 11049701 TI - Educating the infection control team - past, present and future. A British prespective. AB - This review sets out to explore how education and training provisions for members of the Infection Control Team (ICT) have developed alongside their roles and in response to changes in the British National Health Service. It focuses on the Consultant in Communicable Disease Control, the Infection Control Doctor and the Infection Control Nurse in the United Kingdom, but also briefly considers approaches adopted by other countries. Future developments should include maximizing information technology for delivering teaching materials, shared learning and improvements to pre-registration curricula for both doctors and nurses. PMID- 11049702 TI - Selective decontamination of the digestive tract, SDD: a commentary. PMID- 11049703 TI - Molecular investigation of two clusters of hospital-acquired bacteraemia caused by multi-resistant Klebsiella pneumoniae using pulsed-field gel electrophoresis and in frequent restriction site PCR. Infection Control Group. AB - Two molecular typing methods, DNA macrorestriction analysis with XbaI resolved by pulsed-field gel electrophoresis (PFGE) and infrequent restriction site PCR (IRS PCR) assay with adapters designed for XbaI and HhaI restriction sites, were used to investigate two clusters of hospital-acquired bacteraemia associated with multi-resistant Klebsiella pneumoniae which occurred in a paediatric intensive care unit (PICU). A total of 56 K. pneumoniae isolates were analysed. These included 10 bacteraemic isolates from eight patients, 26 isolates obtained during an epidemiological survey, and 20 epidemiologically non-related isolates incorporated as controls. One major pattern was demonstrated in 22 of the 56 isolates analysed. These included nine of the 10 bacteraemic isolates, a single rectal isolate, two hand culture isolates and 10 sink isolates. All of these 22 isolates illustrated identical antibiograms, whilst the other 34 isolates shared six antibiograms and 31 unique patterns by either PFGE or IRS-PCR assay. The two clusters of bacteraemia appeared to be outbreaks induced by the same strain of K. pneumoniae which may have utilized sinks as reservoirs and been transmitted through the hands of medical personnel to patients. IRS-PCR demonstrates concordant results with PFGE analysis in studying the genetic relationships among K. pneumoniae isolates, and serves as an excellent epidemiological tool for this bacterium. PMID- 11049704 TI - Effects on nursing workload of different methicillin-resistant Staphylococcus aureus (MRSA) control strategies. AB - Nursing staff workload may influence hospital-acquired staphylococcal transmission. Closure of wards to new admissions is used in some institutions as part of methicillin-resistant Staphylococcus aureus (MRSA) outbreak control, and we postulated that it worked by reducing staff workload, allowing more time for good infection control practices. We have used the GRASPCopyright workload system to compare nursing workload during six MRSA outbreaks. Two outbreaks occurred while an aggressive control policy ('old'; 1994-1995) was in place, with a low threshold for ward closure. Control measures had been relaxed before the later four outbreaks, with wards remaining fully or partially open unless MRSA transmission proved intractable ('new'; 1995-1996). To standardize the analysis we compared GRASP and epidemiological data for periods while MRSA transmission was occurring on each ward ('during'), and four week periods 'before' and 'after'. Closing wards to admissions reduces staff workload towards a quality environment, although the nursing requirements of remaining patient rises. Workload pressures may rise during outbreaks if wards are not closed quickly and fully, and patients are not transferred to specialist isolation facilities. Changes in nursing workload need to be assessed during comparative studies of outbreak control measures and the GRASP(c) system appears to be a sensitive way to measure these. PMID- 11049705 TI - Control of a methicillin-resistant Staphylococcus aureus outbreak in a neonatal intensive care unit by unselective use of nasal mupirocin ointment. AB - In September 1996, an outbreak of methicillin-resistant Staphylococcus aureus (MRSA) colonization occurred in the neonatal intensive care unit (NICU) of our hospital. After failing to control the outbreak by conventional methods we implemented an intranasal blanket use programme of mupirocin ointment from the beginning of November 1997. In the programme, patients who had been carrying MRSA received intranasal administration of the ointment three times daily for the first three days and consecutively three times weekly, while newly admitted patients and those who had not been colonized were prophylactically medicated three times weekly. This blanket administration was executed for one month. Methicillin-resistant Staphylococcus aureus colonization became undetectable in all but one intubated inpatient who had already been colonized before the start of the programme, and no new acquisitions occurred until the middle of January 1998, seven weeks after the termination of the blanket use programme. The rate of colonized patients in the unit also decreased. During and after the programme, neither an increase in minimum inhibitory concentration for the antibiotic nor apparent adverse reactions in any of the treated patients were observed. We concluded that this procedure is an effective method of controlling an MRSA outbreak in an NICU when the outbreak cannot be managed with conventional measures. PMID- 11049706 TI - Is hospital-acquired intravascular catheter-related sepsis associated with outbreak strains of coagulase-negative staphylococci? AB - Macrorestriction fragment profile analysis by pulsed field gel electrophoresis was used to type strains of coagulase-negative staphylococci (CNS) isolated from 30 patients with catheter-related sepsis at the University Hospital Birmingham NHS Trust, UK. Twenty-three infections were hospital-acquired. A total of 56 CNS were isolated from the patients and identified by API as Staphylococcus epidermidis (54), Staphylococcus lugdunensis (1) and Staphylococcus hominis (1). The micro-organisms were further characterized by antibiograms and restriction digestion using SmaI. Analysis of the macrorestriction fragment profiles demonstrated that the isolates from 24 patients were distinct, whereas a common genotype of S. epidermidis was isolated from the blood cultures of six patients, all of whom had acquired this infection in hospital. Three of these patients were located in a haematology ward, two on an intensive care unit and one on a dialysis unit. The data from this current study suggests that specific strains of S. epidermidis may be an important cause of nosocomial catheter-related sepsis resulting from cross-infection, and that this association would not be detected by conventional typing methods including biotyping and antibiograms. PMID- 11049707 TI - A prospective, randomized, double-blind studyof single high dose versus multiple standard dose gentamicin both in combination withmetronidazole for colorectal surgicalprophylaxis. AB - Single, high dose regimens of gentamicin plus metronidazole for colorectal surgical prophylaxis have not been adequately studied. Patients received single high dose gentamicin (4.5 mg/kg) plus metroni-dazole (500 mg) preoperatively or multiple standard dose gentamicin (1.5 mg/kg) plus metronidazole (500 mg) preoperatively and every 8h for 24h postoperatively. The deep surgical site infection (SSI) rates were 8.1% (6/74) and 6.9% (5/72) in the single high dose and multiple standard dose groups, respectively (P= 0.94). There was a trend towards fewer superficial SSIs in the single high dose group with infection rates of 18.9% (14/74) vs. 30.6% (22/72) (P= 0.05). Diabetes mellitus (odds ratio = 7.04) and surgery duration of longer than 3h (odds ratio = 5.46) were independent risk factors for the development of SSIs. A subset analysis of prolonged operations found significantly fewer superficial SSIs in the single high dose group than in the multiple standard dose group with rates of 22.2% (6/27) vs. 55% (11/20), respectively (P= 0.021). Single high dose gentamicin plus metronidazole preoperatively was at least as effective as the multiple standard dose regimen and may be more effective for prolonged operations. PMID- 11049708 TI - Routine two-step skin testing for tuberculosis in the staff of a geriatric hospital in Israel: booster and conversion rates. AB - The objective of this study was to determine the prevalence of positive skin tests amongst the staff of a 200 bed geriatric hospital in Haifa, Israel. By comparing the findings with those of a study performed five years previously, we hoped to ascertain the number of conversions which had occurred in the period studied. This was undertaken in order to assess a new policy from the Israel Ministry of Health regarding skin testing for health care workers. We also hoped to decide upon the frequency of skin testing required and to compare data from recent immigrants from countries with a high prevalence of TB. In 1997, we performed two-step skin testing (TSST) on 318 health care workers. We ascertained the number of positive reactions on the first and second testing and calculated the number of subjects who showed significant boosting. We also compared the results to those obtained in a study in 1992 and calculated the rate of conversion. We used multivariate analysis to examine the effects of age, gender, country of origin, years in Israel, previous BCG vaccination, previous exposure to contagious TB, work site and area of residence in the city, on the response to TSST. Between 1990 and 1996, 655 000 immigrants from the former USSR arrived; 'recent immigration' was defined from that date onward. The final number of positive reactions out of 282 subjects, who were either positive or negative on step 1 and presented for step 2, was 171 (60%). Booster effect was not significantly associated with any of the variables examined. The size of reaction in TSST was related to country of origin and recent immigration. The 83 recent immigrants from the former USSR had more frequent (61%) and larger reactions (mean (sd): 9.0 (6.46) mm) than the 114 native-born Israelis with 39% positive reactions (6.2 (5.89) P= 0.009). Comparison with 1992 revealed 26 (31%) of previous negatives as positive. Conversion was associated with age. All conversions save one were in individuals younger than 50 years (P= 0.07). In conclusion, TSST, performed to enable detection of recent infection after exposure to contagious TB, was relevant for 40% of health care workers (HCWs). Second step testing contributed an additional 23% positive reactions. New immigrants had larger initial reactions. Conversion occurred mostly in younger workers and could be either due to unrecognized TB in the hospital or to exposure in the community. PMID- 11049709 TI - Evaluation of chlorhexidine and povidone iodine activity against methicillin resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecalis using a surface test. AB - Most published studies of the activity of biocides against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) have been based on suspension tests. This study was undertaken to provide information on the effect of chlorhexidine and povidone iodine on bacteria dried on to surfaces, a situation in which biocide activity is known to be reduced. The inactivation of MRSA (10 strains), methicillin-sensitive Staphylococcus aureus (MSSA, 10 strains), VRE (nine strains) and vancomycin-sensitive Enterococcus faecalis (VSE, 10 strains) by 0.5% aqueous chlorhexidine gluconate or 10% povidone iodine was evaluated by applying the European surface test method. Povidone iodine was equally active against resistant and sensitive strains of both species with microbicidal effects (ME), i.e. the log(10)concentration of micro-organisms compared with controls treated with distilled water, after 1.5 min of 3.14 and 3.49 for VRE and VSE respectively, and 3.47 and 3.78 for MRSA and MSSA. Chlorhexidine was equally active against VRE and VSE (ME 3.37 vs. 3. 56 after 7 min, respectively), but was significantly less active against MRSA as opposed to MSSA (ME 3.07 vs. 3.83 after 10 min, P= 0. 017). PMID- 11049710 TI - The effect of super-oxidized water on Escherichia coli. AB - The mechanism of action of Sterilox, a non-toxic liquid biocide produced by electrolysis of a dilute saline solution, upon planktonic cells of Escherichia coli JM109 was investigated using protein and nucleic acid analysis. The results revealed total destruction of chromosomal and plasmid DNA, RNA and proteins of E. coli within 5 min of exposure. Our earlier investigation conducted using atomic force microscopy imaging revealed swelling and rupture of E. coli cells with release of cytoplasm. We propose that the biocidal properties of Sterilox are due to its effect upon constituents of the bacterial cell including proteins and nucleic acids. PMID- 11049711 TI - Strain identities of phage non-typable MRSA in the UK. PMID- 11049712 TI - Methicillin-resistant Staphylococcus aureus in a Delhi teaching hospital. PMID- 11049713 TI - Salmonella group B meningitis six weeks after hospitalization in a neonatal care unit. PMID- 11049716 TI - Preface. PMID- 11049714 TI - Maintenance of peripheral and central intravenous infusion devices by 0.9% sodium chloride with or without heparin as a potential source of catheter microbial contamination. PMID- 11049728 TI - Substance P: an historical perspective. PMID- 11049729 TI - Update on Substance P (NK-1 receptor) antagonists in clinical trials for depression. PMID- 11049730 TI - Neuroanatomical localisation of Substance P in the CNS and sensory neurons. AB - The anatomical distribution of Substance P (SP) has been investigated since the development of antibodies against it in the 1970s. Although initial studies were performed with antibodies that also recognised the other endogenous neurokinins, most of the initial descriptions are surprisingly still valid today. In this review, we provide an integrated overview of the pathways containing SP in the central and peripheral nervous systems. The highest densities of SP immunoreactivity occur in the superficial dorsal horn of the spinal cord, in the substantia nigra and in the medial amygdaloid nucleus. In the peripheral nervous system, SP occurs in high concentrations in small diameter primary sensory fibres and in the enteric nervous system. SP is extensively co-localised with classical transmitters and other neuropeptides. In the spinal cord, SP immunoreactive axonal boutons are preferentially presynaptic to neurons expressing the SP receptor, suggesting that the neurokinin acts at a short distance from the release site. In contrast, in the periphery, the situation probably differs in the autonomic ganglia, where the targets are directly innervated by SP, and in other peripheral territories, where SP has to diffuse through the connective tissue to reach the structures expressing the receptor. PMID- 11049731 TI - Substance P and its role in neural mechanisms governing learning, anxiety and functional recovery. AB - The neurokinin Substance P (SP) is widely distributed in the central nervous system and has been extensively studied in various functional aspects. This review focuses on the behavioral relevance of SP. Here we show that SP can have memory-promoting, reinforcing and anxiolytic-like effects when administered systemically or into the nucleus basalis of the ventral pallidum. These effects seem to be mediated via the SP-preferring NK(1)receptor and differentially related to N- versus C-terminal fragments of the undecapeptide. Secondly, SP injection into the ventral pallidum can lead to increases of acetylcholine in frontal cortex and dopamine in nucleus accumbens, suggesting that the hypermnestic, positively reinforcing and anxiolytic effects observed upon basal forebrain injection of SP are mediated by activation of the nucleus accumbens ventral pallidum circuitry. Furthermore, SP and certain SP-fragments may not only be considered to have beneficial behavioral effects in normal animals, but can also prevent lesion-induced functional deficits and improve the speed of recovery. This indicates that SP agonists might also have a neuroprotective capacity in parallel with recovery-promoting actions. PMID- 11049732 TI - NGF over-expression during development leads to permanent alterations in innervation in the spinal cord and in behavioural responses to sensory stimuli. AB - Transgenic mice expressing nerve growth factor (NGF) under the control of a myelin basic protein promoter display above normal NGF levels in the spinal white matter from birth to the age of 2 months. These transient high levels of NGF result in a lasting hyper-innervation of the spinal white matter by ectopic Substance P (SP)-immunoreactive (IR) sensory fibres. Ultrastructural studies in adult transgenic mice demonstrated that the SP-containing fibres establish synapses on neuronal dendrites in the white matter and that most such dendrites express SP receptors. The transgenic animals display a stimulus-induced hyperalgesia and allodynia in a test measuring the latency to tail withdrawal following a heat stimulus. The hyperalgesia and allodynia were reversed by systemic administration of SP receptor or NMDA receptor antagonists. Surprisingly, the application of morphine resulted in an increase in withdrawal latency which was greater than that observed in non-transgenic controls. PMID- 11049733 TI - Molecular models to analyse preprotachykinin-A expression and function. AB - Towards an understanding of the mechanisms controlling Preprotachykinin A (PPT) expression we have generated a variety of molecular models to determine the mechanisms regulating both the tissue-specific and stimulus-inducible expression of the PPT gene. The approaches used include transgenic and virus vector models complementing biochemical analysis of promoter interactions with transcription factors. We have identified and characterised a yeast artificial chromosome (YAC) containing the human PPT gene and generated transgenic mouse lines containing multiple copies of this chromosome on a normal mouse genetic background. This resulted in a pattern of expression in the nervous system remarkably similar to that reported for PPT mRNA in rodents. In addition, this transgenic model has been constructed in such a manner to allow for over expression of tachykinins based on the number of extra alleles in the transgenic mouse. These animals allow us to further examine the function of the tachykinins and acts as a useful complement to existing PPT ablated mice. In vitro we have introduced the proximal PPT promoter in reporter gene constructs into adult neurones in both DRG and the CNS by an adenoassociated virus (AAV) vector or by biolistic transfection respectively. Using the AAV vector we have demonstrated that the proximal promoter can mediate the effects of NGF in adult rat DRG. These models allow us to delineate transcriptional domains involved in the physiological and pathological expression of the PPT gene. PMID- 11049734 TI - Peripheral actions of tachykinins. AB - Tachykinins mediate a variety of physiological processes in the gastrointestinal, pulmonary and genito-urinary tract mainly through the stimulation of NK1 and NK2 receptors. Preclinical evidence obtained through the use of selective tachykinin receptor antagonists indicates that endogenous tachykinins are involved in augmented smooth muscle contraction, vasodilatation, chemotaxis and activation of immune cells, mucus secretion, water absorption/secretion. Recent evidence also suggests that endogenous tachykinins released at the peripheral level may play a role in visceral inflammation, hyperreflexia and hyperalgesia. Possible mechanisms underlying the stimulation of primary afferent neurons by tachykinins may involve a direct excitation of these neurons and the release of mediators which sensitise or stimulate sensory nerves. Tachykinin receptor antagonists could have a clinical utility in several human diseases such as irritable bowel syndrome, asthma, and in micturition disturbances characterized by a hyperactive bladder. PMID- 11049735 TI - Neurokinin mediation of edema and inflammation. AB - The aim of this article is to furnish a brief review of the role played by neurokinins in the inflammatory process. Further attention is given to the mechanisms, as well as to the receptor subtypes involved in neurokinin-mediated inflammation, in an attempt to clarify the participation of neurokinins in different models of acute and chronic inflammation. The involvement of SP, NKA and NKB is also examined in relation to the major signs of inflammation, including edema formation, protein plasma extravasation and vasodilatation. Finally, we provide a general overview on the potential clinical applications of neurokinin antagonists, along with the involvement of neurokinins in human diseases. PMID- 11049736 TI - Entering through the doors of perception: characterization of a highly selective Substance P receptor-targeted toxin. AB - Perception of external stimuli is often mediated through the activity of a G protein-coupled receptor in response to its ligand. Receptor-mediated internalization allows the insertion of toxins that cause the elimination of receptor-expressing neurons. Using this technique new information on systems biology can be discovered and with this, new therapeutics developed. PMID- 11049737 TI - Factor X fusion proteins: improved production and use in the release in vitro of biologically active hirudin from an inactive alpha-factor-hirudin fusion protein. AB - Many recombinant proteins are synthesized as fusion proteins containing affinity tags to aid in the downstream processing. After purification, the affinity tag is often removed by using a site-specific protease such as factor Xa (FXa). However, the use of FXa is limited by its expense and availability from plasma. To develop a recombinant source of FXa, we have expressed two novel forms of FXa using baby hamster kidney (BHK) cells as host and the expression vector pNUT. The chimeric protein FIIFX consisted of the prepropeptide and the Gla domain of prothrombin linked to the activation peptide and protease region of FXa, together with a cellulose-binding domain (CBD(Cex)) as an affinity tag. A second variant consisted of the transferrin signal peptide linked to the second epidermal growth factor-like domain and the catalytic domain of FX and a polyhistidine tag. Both FX variants were secreted into the medium, their affinity tags were functional, and following activation, both retained FXa-specific proteolytic activity. However, the yield of the FIIFX-CBD(Cex) fusion protein was 10-fold higher than that of FX-CBD(Cex) and other forms of recombinant FX reported to date. The FXa derivatives were used to cleave two different fusion proteins, including a biologically inactive alpha-factor-hirudin fusion protein secreted by Saccharomyces cerevisiae. After cleavage, the released hirudin demonstrated biological activity in a thrombin inhibition assay, suggesting that this method may be applicable to the production of toxic or unstable proteins. The availability of novel FX derivatives linked to different affinity tags allows the development of a versatile system for processing fusion proteins in vitro. PMID- 11049738 TI - Purification and characterization of the highly thermostable proteases from Bacillus stearothermophilus TLS33. AB - Three thermostable proteases, designated S, N, and B, are extracellular enzymes produced by Bacillus stearothermophilus strain TLS33. They were purified by lysine affinity chromatography, strong anion exchange Q HyperD chromatography, and Ultrogel AcA44 gel filtration. The molecular masses of the enzymes determined by SDS-PAGE and zymography were approximately 36, 53, and 71 kDa, respectively. Thermostable protease S bound strongly to the lysine affinity column and could be purified by this single step. The optimum pH values of proteases S, N, and B were shown to be 8.5, 7.5, and 7.0, respectively. The maximum activities for the enzymes were at 70, 85, and 90 degrees C, respectively. Proteases S, N, and B at pH 7.0 in the presence of 5 mM CaCl(2) retained half their activities after 30 min at 72, 78, and 90 degrees C, respectively. All three thermostable proteases were strongly inhibited by the metal chelators EDTA and 1,10-phenanthroline, and the proteolytic activities were restored by addition of ZnCl(2). They can thus be classified as Zn(2+) metalloproteases. The cleavage specificities of proteases S, N, and B on a 30-residue synthetic peptide from pro-BPN' subtilisin were Tyr-Ile, Phe-Lys, and Gly-Phe, respectively. PMID- 11049739 TI - Large-scale expression, refolding, and purification of the catalytic domain of human macrophage metalloelastase (MMP-12) in Escherichia coli. AB - We have cloned, overexpressed, and purified the catalytic domain (residues Gly106 to Asn268) of human macrophage metalloelastase (MMP-12) in Escherichia coli. This construct represents a truncated form of the enzyme, lacking the N-terminal propeptide domain and the C-terminal hemopexin-like domain. The overexpressed protein was localized exclusively to insoluble inclusion bodies, in which it was present as both an intact form and an N-terminally truncated form. Inclusion bodies were solubilized in an 8 M guanidine-HCl buffer and purified by gel filtration chromatography under denaturing conditions. Partial refolding of the protein by dialysis into a 3 M urea buffer caused selective degradation of the truncated form of the protein, while the intact catalytic domain was unaffected by proteolysis. An SP-Sepharose chromatography step purified the protein to homogeneity and served also to complete the refolding. The purified protein was homogeneous by mass spectrometry and had an activity similar to that of the recombinant enzyme purified from mammalian cells. The protein was both soluble and monodisperse at a concentration of 9 mg/ml. This purification procedure enables the production of 23 mg of protein per liter of E. coli culture and is amenable to large-scale protein production for structural studies. PMID- 11049740 TI - Purification of alpha-amylase isoenzymes from Scytalidium thermophilum on a fluidized bed of alginate beads followed by concanavalin A-agarose column chromatography. AB - An alpha-amylase has been purified from the thermophilic fungus Scytalidium thermophilum. A ninefold purification was achieved in a single step using fluidized bed chromatography wherein alginate was used as the affinity matrix. There are at least two isoenzymes as shown by concanavalin A (Con A)-agarose column chromatography. The isoenzyme binding to Con A is stable for at least 3 h at 80 degrees C in the presence of calcium ions. The isoenzymes have similar molecular weights of around 45,000 Da as shown by SDS-PAGE analysis. The isoenzymes differ only slightly in their pH optima and temperature optima but the isoenzyme binding to Con A-agarose has slightly higher thermal stability. PMID- 11049741 TI - Purification of recombinant Arabidopsis thaliana dehydrins by metal ion affinity chromatography. AB - In this study we describe a novel method for purification of Arabidopsis thaliana dehydrins overproduced in Escherichia coli. The cDNAs corresponding to the four dehydrin genes RAB18, LTI29, LTI30, and COR47 were inserted into a bacterial expression vector under an isopropyl beta-d-thiogalactopyranoside (IPTG) inducible bacterial promoter. After IPTG induction all four proteins accumulated in high amounts. The recombinant proteins were efficiently purified to over 95% purity with a three-step purification scheme: heat fractionation, immobilized metal ion affinity chromatography (IMAC), and ion exchange chromatography. In this study we introduce the novel use of IMAC as an efficient purification method for native dehydrins. Characterization of the purified proteins was done by Edman degradation, mass spectrometry, reverse-phase chromatography, and analytical gel filtration under native and denaturing conditions. Yields of purified proteins were between 2.8 and 12.5 mg per liter of bacterial culture, sufficient for further biochemical studies. PMID- 11049742 TI - Glycosylation of prourokinase produced by Pichia pastoris impairs enzymatic activity but not secretion. AB - Both glycosylated and nonglycosylated forms of recombinant human prourokinase were produced to the level of 20 mg/L by yeast Pichia pastoris in BMMY medium after 2 days of culture. The expressed pro-UK was 98% secreted into the culture medium and easily purified by carboxymethyl cellulose chromatography. More than 99% of pro-UK in the culture medium was found in single-chain form. This was contradictory to a previous finding which found that glycosylation of pro-UK by yeast inhibited its secretion. The absence of glycosylation at Asn302 of pro-UK has no measurable effect on its secretion from the yeast cells. However, the nonglycosylated pro-UK was much less stable in the culture medium, probably due to proteolysis. Nonglycosylated pro-UK from yeast had a clot lysing activity comparable to that of Escherichia coli-derived or mammalian cell-derived recombinant pro-UK. By contrast, the glycosylated yeast pro-UK was less activatable by plasmin and had a lower enzymatic activity against plasminogen and a lower clot lysing activity than nonglycosylated pro-UK from yeast, while their amidolytic activity against S2444 was equivalent. It was concluded that glycosylation of pro-UK by yeast P. pastoris interferes with the catalytic site but not secretion of this protein. PMID- 11049743 TI - Expression, purification, refolding, and characterization of recombinant human interleukin-13: utilization of intracellular processing. AB - Interleukin-13 (IL-13) is a pleiotropic cytokine that elicits both proinflammatory and anti-inflammatory immune responses. Recent studies underscore its role in several diseases, including asthma and cancer. Solution studies of IL 13 and its soluble receptors may facilitate the design of antagonists/agonists which would require milligram quantities of specifically labeled protein. A synthetic gene encoding human IL-13 (hIL-13) was inserted into the pMAL-c2 vector with a cleavage site for the tobacco etch virus (TEV) protease. Coexpression of the fusion protein and TEV protease led to in vivo cleavage, resulting in high levels of hIL-13 production. hIL-13, localized to inclusion bodies, was purified and refolded to yield approximately 2 mg per liter of bacteria grown in minimal media. Subsequent biochemical and biophysical analysis of both the unlabeled and (15)N-labeled protein revealed a bioactive helical monomer. In addition, the two disulfide bonds were unambiguously demonstrated to be Cys29-Cys57 and Cys45-Cys71 by a combined proteolytic digestion and mass spectrometric analysis. PMID- 11049744 TI - Expression and purification of an active, full-length hepatitis C viral NS4A. AB - The nonstructural protein 3 (NS3) of the hepatitis C virus (HCV) is a bifunctional protein with protease and helicase activities. Nonstructural protein 4A (NS4A) is preceded by NS3 and augments the proteolytic activity of NS3 through protein-protein interaction. The central domain of NS4A has been shown to be sufficient for the enhancement of the NS3 protease activity. However, investigations on the roles of the N-terminal and the C-terminal regions of NS4A have been hampered by the difficulty of purification of full-length NS4A, a polypeptide that contains highly hydrophobic amino acid residues. Here we report a procedure by which one can produce and purify an active, full-length NS4A using maltose-binding protein fusion method. The full-length NS4A fused to the maltose binding protein is soluble and maintains its NS3 protease-enhancing activity. PMID- 11049745 TI - Expression and purification of mammalian calreticulin in Pichia pastoris. AB - Calreticulin is a 46-kDa Ca(2+)-binding chaperone of the endoplasmic reticulum membranes. The protein binds Ca(2+) with high capacity, affects intracellular Ca(2+) homeostasis, and functions as a lectin-like chaperone. In this study, we describe expression and purification procedures for the isolation of recombinant rabbit calreticulin. The calreticulin was expressed in Pichia pastoris and purified to homogeneity by DEAE-Sepharose and Resource Q FPLC chromatography. The protein was not retained in the endoplasmic reticulum of Pichia pastoris but instead it was secreted into the external media. The purification procedures reported here for recombinant calreticulin yield homogeneous preparations of the protein by SDS-PAGE and mass spectroscopy analysis. Purified calreticulin was identified by its NH(2)-terminal amino acid sequences, by its Ca(2+) binding, and by its reactivity with anti-calreticulin antibodies. The protein contained one disulfide bond between (88)Cys and (120)Cys. CD spectral analysis and Ca(2+) binding properties of the recombinant protein indicated that it was correctly folded. PMID- 11049746 TI - Large-scale purification of recombinant human angiostatin. AB - A process for the purification of recombinant human angiostatin (rhAngiostatin), produced by Pichia pastoris fermentation operated at the 2000-L scale, is reported. rhAngiostatin was recovered and purified directly from crude fermentation broth by cation exchange expanded bed adsorption chromatography. Anion exchange chromatography, hydroxyapatite chromatography, and hydrophobic interaction chromatography were used for further purification. Full-length rhAngiostatin was separated from rhAngiostatin molecules fragmented by endoproteolysis. On average, 140 g of rhAngiostatin was produced per batch, with an overall yield of 59% (n = 9). The purification process was completed in approximately 48 h and used only inexpensive and nontoxic raw materials. Methods development, process synthesis, and process scale-up data are presented and discussed. PMID- 11049747 TI - Purification and characterization of human alpha-galactosidase A expressed in insect cells using a baculovirus vector. AB - Fabry disease is an X-linked inborn error of glycolipid metabolism caused by deficiency of the lysosomal enzyme alpha-galactosidase A. The enzyme is responsible for the hydrolysis of terminal alpha-galactoside linkages in various glycolipids. To perform more extensive biochemical characterization and to develop new approaches for enzyme therapy, a method of producing and purifying recombinant alpha-galactosidase A suitable for scale-up manufacture for use in humans is needed. Previously, a catalytically active recombinant human alpha galactosidase A was expressed using a baculovirus vector and purified using conventional chromatography. However, the level of expression was too low to permit economical production and the chromatographic techniques used for enzyme purification were not suitable for enzyme to be used in humans. Therefore, the cDNA of the enzyme was cloned to an improved baculovirus vector and the enzyme was expressed in a 15-liter bioreactor using optimized growth conditions. Infection of insect cells by the baculovirus resulted in a significant fivefold increase in the level of secreted recombinant alpha-galactosidase A activity that is compatible with economic manufacturing. The recombinant alpha-galactosidase A was purified to homogeneity using ion exchange (Poros 20-CM, Poros 20-HQ) and hydrophobic chromatography (Toso-ether, Toso-butyl) using the BioCAD HPLC workstation. These chromatographic steps are readily scalable to larger volumes and are appropriate for the purification of the recombinant human alpha galactosidase A to be used in clinical trials of enzyme replacement therapy for Fabry disease patients. PMID- 11049748 TI - Expression, purification, and characterization of the protein repair l isoaspartyl methyltransferase from Arabidopsis thaliana. AB - Protein l-isoaspartate (d-aspartate) O-methyltransferase (EC 2.1.1. 77) is a repair enzyme that methylates abnormal l-isoaspartate residues in proteins which arise spontaneously as a result of aging. This enzyme initiates their conversion back into the normal l-aspartate form by a methyl esterification reaction. Previously, partial cDNAs of this enzyme were isolated from the higher plant Arabidopsis thaliana. In this study, we report the cloning and expression of a full-length cDNA of l-isoaspartyl methyltransferase from A. thaliana into Escherichia coli under the P(BAD) promoter, which offers a high level of expression under a tight regulatory control. The enzyme is found largely in the soluble fraction. We purified this recombinant enzyme to homogeneity using a series of steps involving DEAE-cellulose, gel filtration, and hydrophobic interaction chromatographies. The homogeneous enzyme was found to have maximum activity at 45 degrees C and a pH optimum from 7 to 8. The enzyme was found to have a wide range of affinities for l-isoaspartate-containing peptides and displayed relatively poor reactivity toward protein substrates. The best methyl accepting substrates were KASA-l-isoAsp-LAKY (K(m) = 80 microM) and VYP-l-isoAsp HA (K(m) = 310 microM). We also expressed the full-length form and a truncated version of this enzyme (lacking the N-terminal 26 amino acid residues) in E. coli under the T7 promoter. Both the full-length and the truncated forms were active, though overexpression of the truncated enzyme led to a complete loss of activity. PMID- 11049749 TI - High-level periplasmic expression in Escherichia coli using a eukaryotic signal peptide: importance of codon usage at the 5' end of the coding sequence. AB - We investigated the ability of signal peptides of eukaryotic origin (human, mouse, and yeast) to efficiently direct model proteins to the Escherichia coli periplasm. These were compared against a well-characterized prokaryotic signal peptide-OmpA. Surprisingly, eukaryotic signal peptides can work very efficiently in E. coli, but require optimization of codon usage by codon-based mutagenesis of the signal peptide coding region. Analysis of the 5' of periplasmic and cytoplasmic E. coli genes shows some codon usage differences. PMID- 11049750 TI - High-level expression of biologically active human prolactin from recombinant baculovirus in insect cells. AB - We examined the feasibility of high-level production of recombinant human prolactin, a multifunctional protein hormone, in insect cells using a baculovirus expression system. The human prolactin cDNA with and without the secretory signal sequence was cloned into pFastBac1 baculovirus vector under the control of polyhedrin promoter. Prolactin was produced upon infection of either Sf9 or High Five cells with the recombinant baculovirus containing the human prolactin cDNA. The production of recombinant prolactin varied from 20 to 40 mg/L of monolayer culture, depending on the cell types. The prolactin polypeptide with its own secretory signal was secreted into the medium. N-terminal amino acid sequence analysis of the recombinant polypeptide purified from the culture medium indicated that the protein was processed similar to human pituitary prolactin. Carbohydrate analysis of the purified protein indicated that a fraction of the recombinant prolactin made in insect cells appeared to be glycosylated. Also, both secreted and nonsecreted forms of the recombinant prolactin in insect cells were biologically equivalent to the native human prolactin (pituitary derived) in the Nb2 lymphoma cell proliferation assay. PMID- 11049751 TI - Chemical and posttranslational modification of Escherichia coli acyl carrier protein for preparation of dansyl-acyl carrier proteins. AB - Escherichia coli acyl carrier protein (ACP) contains a single tyrosine residue at position 71. The combined o-nitration of apo-ACP Y71 by tetranitromethane and reduction to 3-aminotyrosyl-apo-ACP were performed to introduce a specific site for attachment of a dansyl fluorescent label. Conditions for purification and characterization of dansylaminotyrosyl-apo-ACP are reported. Dansylaminotyrosyl apo-ACP was enzymatically phosphopantetheinylated and acylated in vitro with an overall approximately 30% yield of purified stearoyl-dansylaminotyrosyl-ACP starting from unmodified apo-ACP. The steady-state kinetic parameters k(cat) = 22 min(-1) and K(M) = 2.7 microM were determined for reaction of stearoyl dansylaminotyrosyl-ACP with stearoyl-ACP Delta(9)-desaturase. These results show that dansylaminotyrosyl-ACP will function well for studying binding interactions with the Delta(9)-desaturase and suggest similar possibilities for other ACP dependent enzymes. The efficient in vivo phosphopantetheinylation of E. coli apo ACP by coexpression with holo-ACP synthase in E. coli BL21(DE3) using fructose as the carbon source is also reported. PMID- 11049752 TI - Characterization of a recombinant murine 18.5-kDa myelin basic protein. AB - A recombinant hexahistidine-tagged 18.5-kDa isoform of murine myelin basic protein has been characterized biochemically and immunogenically, by mass spectrometry, by circular dichroism under various conditions (in aqueous solution, with monosialoganglioside G(M1), and in 89% 2-propanol), and by transmission electron microscopy. The preparations of this protein indicated a high degree of purity and homogeneity, with no significant posttranslational modifications. Circular dichroic spectra showed that this preparation had the same degree of secondary structure as the natural bovine 18.5-kDa isoform of myelin basic protein. Incubation of the recombinant protein with lipid monolayers containing a nickel-chelating lipid resulted in the formation of fibrous assemblies that formed paracrystals of spacings 4.8 nm between fibers and 3-4 nm along them. PMID- 11049753 TI - Analysis of three overexpression systems for VanX, the Zinc(II) dipeptidase required for high-level vancomycin resistance in bacteria. AB - The gene from Enterococcus faecilis encoding the dipeptidase VanX was subcloned into overexpression vectors pET-5b, pET-27b, and IMPACT-T7, and VanX was overexpressed in BL21(DE3) pLysS Escherichia coli. The pET-5b-vanx overexpression plasmid produces VanX at approximately 12 mg/L under optimum conditions. VanX produced from this overexpression system exists primarily as a dimer in solution, binds ca. 1 Zn(II) ion per monomer, and exhibits K(m) and k(cat) values of 500 +/ 40 microM and 0.074 +/- 0.001 s(-1), respectively, when l-alanine-p-nitroanilide is used as substrate. The IMPACT-T7-vanx overexpression plasmid produces a VanX fusion protein with a chitin-binding domain that allows for purification of the fusion construct with a chitin column. Cleavage of the fusion protein is completed by an on-column chemical cleavage, resulting in approximately 10 mg/L of purified VanX. VanX produced from this system is identical to that produced from the pET-5b system, except the CD spectrum of the IMPACT-T7-produced VanX suggests a small change in secondary structure. This change in secondary structure does not affect any of the kinetic or metal-binding properties of the enzyme. The pET-27b-vanx overexpression plasmid produces and secretes VanX into the growth medium; this system allows for 20 mg of VanX to be isolated per liter of growth medium. The pET-27b-produced VanX is identical to that produced from pET-5b. PMID- 11049754 TI - Recombinant expression of the Candida rugosa lip4 lipase in Escherichia coli. AB - It is difficult to express recombinant Candida rugosa lipases (CRLs) in heterologous systems, since C. rugosa utilizes a nonuniversal serine codon CUG for leucine. In this study, recombinant LIP4 in which all 19 CUG codons had been converted to a universal serine codon was overexpressed in Escherichia coli BL21(DE3). The recombinant LIP4 was found mainly in the inclusion bodies and showed a low catalytic activity. To increase the amount of soluble form and activity of recombinant LIP4, the DNA was fused to the gene for thioredoxin (TrxFus-LIP4) and then expressed in E. coli strain AD494(DE3). This strategy promotes the formation of disulfide bonds in the cytosol and yields enzymatically active forms of LIP4. The purified recombinant TrxFus-LIP4 and LIP4 expressed in AD494(DE3) had the same catalytic profiles. In addition, recombinant LIP4 had higher esterase activities toward long-chain ester and lower lipase activities toward tributyrin, triolein, and olive oil. This system for the expression of fungal lipase in E. coli strain AD494(DE3) is reliable and may produce enzymatically active forms of recombinant lipase without an in vitro refolding procedure. PMID- 11049755 TI - Expression and purification of dynamin II domains and initial studies on structure and function. AB - Dynamin II, a large GTP-binding protein, is involved in endocytosis and in vesicle formation at the trans-Golgi network. To further elucidate functions of dynamin II, the pleckstrin homology domain (PHD), the proline-rich domain (PRD), and the C-terminal part of dynamin II (dynamin(500-870)) were expressed in Escherichia coli. The PHD, tagged C-terminally by a (His)(6) peptide, was expressed to 15% of cellular proteins and could be purified on nickel-chelating agarose. On the contrary, the PRD and dynamin(500-870) had to be tagged with a (His)(6) peptide at the N-terminus to bind to nickel-chelating agarose. Additional tagging with the S-peptide, which forms a stable complex with immobilized S-protein, allowed removal of strongly interacting E. coli proteins. Circular dichroic spectra indicate a structured recombinant PHD with a secondary structure content similar to that of the known PHD from dynamin I. The N terminally tagged, recombinant PRD is unfolded but nevertheless binds specifically to the SH3 domain of amphiphysin II as well as to proteins extracted from rat brain. The described methods are suitable to isolate functionally active domains of dynamin II in sufficient amount and purity for further studies. PMID- 11049756 TI - Bioreactor-scale production and one-step purification of Epstein-Barr nuclear antigen 1 expressed in baculovirus-infected insect cells. AB - Epstein-Barr virus (EBV)-encoded nuclear antigen 1 (EBNA1) is expressed in all EBV-associated malignancies and is essential for EBV-genome maintenance. Antibodies to EBNA1 are abundantly detected in serum of most EBV carriers but EBNA1 escapes recognition by effector T-lymphocytes. To further study the functional and immunological characteristics of EBNA1 it is important to have sufficient quantities of purified EBNA1 available. This paper describes a simple, reproducible method for the production and purification of EBV-encoded EBNA1 expressed in insect cells (bEBNA1). For quantification of EBNA1 expression levels in cell lines and for monitoring bEBNA1 purification and overall yields we developed a quantitative and EBNA1-specific capture ELISA. We observed that EBV positive cell lines express EBNA1 at different levels, with the B cell lymphoblastoid cell line X50/7 having the highest production. However, much larger quantities (380-fold) were obtained by expressing bEBNA1 in recombinant baculovirus-infected Sf9 insect cells. Scaling-up experiments revealed that bEBNA1 expression kinetics and protein stability are identical in 1-liter stirred bioreactors when compared to expression in stationary culture flasks. Optimal expression was reached after 72 h following inoculation at 1 pfu/cell, when insect cell viability was about 50%. For purification the nuclear fraction containing most of the bEBNA1 (>95%) was isolated. Solubilized bEBNA1 was purified by a one-step oriP DNA-Sepharose affinity purification procedure, using biotinylated PCR-amplified family of repeats (FR)-domain products immobilized onto streptavidin agarose. A >200-fold specific enrichment was reached and yields of bEBNA1 with an estimated purity of >95%. PMID- 11049757 TI - High-level expression of human liver monoamine oxidase B in Pichia pastoris. AB - The high-level heterologous expression, purification, and characterization of the mitochondrial outer membrane enzyme human liver monoamine oxidase B (MAO B) using the methylotrophic yeast Pichia pastoris expression system are described. A 2-L culture of P. pastoris expresses approximately 1700 U of MAO B activity, with the recombinant enzyme associated tightly with the membrane fraction of the cell lysate. By a modification of the published procedure for purification of bovine liver MAO B [Salach, J. I. (1979) Arch. Biochem. Biophys. 192, 128-137], recombinant human liver MAO B is purified in a 34% yield ( approximately 200 mg from 2 L of cell culture). The isolated enzyme exhibits an M(r) of approximately 60, 000 on SDS-PAGE and 59,474 from electrospray mass spectrometry measurements, which is in good agreement with the mass predicted from the gene sequence and inclusion of the covalent FAD. One mole of covalent FAD per mole of MAO B is present in the purified enzyme and is bound by an 8alpha-S-cysteinyl(397) linkage, as identified by electrospray mass spectrometry of the isolated tryptic/chymotryptic flavin peptide. Recombinant human liver MAO B and bovine liver MAO B are shown to be acetylated at the seryl residues at their respective amino termini. The benzylamine oxidase activity of recombinant MAO B ranges from 3.0 to 3.4 U/mg and steady-state kinetic parameters for this enzyme preparation compare well with those published for the bovine liver enzyme: k(cat) = 600 min( 1), K(m)(benzylamine) = 0.50 mM, and K(m)(O(2)) = 0.33 mM. Kinetic isotope effect parameters using [alpha,alpha-(2)H(2)]benzylamine are also similar to those found for the bovine enzyme. Recombinant MAO B exhibits a (D)k(cat) = 4.7, a (D)[k(cat)/K(m)(benzylamine)] = 4.5, and a (D)[k(cat)/K(m)(O(2))] = 1.0. In contrast to bovine liver MAO B, no evidence was found for the presence of any anionic flavin radical either by UV-vis or by EPR spectroscopy in the resting form of the enzyme. These data demonstrate the successful heterologous expression of a functional, membrane-bound MAO B, which will permit a number of mutagenesis studies as structural and mechanistic probes not previously possible. PMID- 11049761 TI - What are the microbial components implicated in the pathogenesis of sepsis? Report on a symposium. AB - Despite considerable efforts in the past quarter century to improve therapy for sepsis, mortality rates remain unacceptably high. Microbe-derived constituents can induce the host to produce many mediators that can contribute to immune dysregulation, tissue damage, and death. Although endotoxin-mediated events are clearly important in gram-negative infections, gram-positive bacteria can also play a dominant role. Understanding the interplay of microbial constituents and host immune or inflammatory responses prompted a meeting at Rockefeller University in May 1998. Participants discussed the relative merits of a "2-hit" hypothesis to explain the course of lethal septic shock and a "multihit" synergistic threshold hypothesis. Recommendations include the following: (1) developing animal models that closely mimic human sepsis; (2) further investigating antibiotic effects on bacteria; (3) assessing the relationships between endotoxin, prokaryotic DNA, and peptidoglycan (i.e., independent, additive, or synergistic) in inducing host responses; and (4) developing new strategies to improve outcomes. Studies are needed to better define which and how different microbial constituents lead to sepsis and to provide critical leads for therapeutic intervention. PMID- 11049762 TI - Management of invasive pulmonary aspergillosis in hematology patients: a review of 87 consecutive cases at a single institution. AB - Eighty-seven patients with hematologic malignancies and invasive pulmonary aspergillosis (IPA) were identified between 1982 and 1995. Of these, 39 underwent lung resection on the basis of radiological detection of at least 1 lesion with imaging suggestive of aspergillosis (LISA). IPA was confirmed histologically in 35. The presence of LISA had 90% positive predictive value for IPA. The actuarial survival at 2 years was 36% for 37 patients treated surgically, 20% for 12 patients with unresected LISA but no cultures of Aspergillus species, and 5% for 21 patients diagnosed only by isolation of Aspergillus from respiratory secretions. Analysis by proportional hazard models showed a significant independent negative association between the radiological appearance of LISA and death from all causes. Relapsed hematologic disease was independently significantly associated with death. Age, sex, surgery, previous bone marrow transplantation, or Aspergillus isolation were not independent predictors of death. IPA presenting as LISA carries a relatively good prognosis, possibly explaining the better survival of patients undergoing surgery for such lesions. PMID- 11049763 TI - Prospective study of the usefulness of sputum Gram stain in the initial approach to community-acquired pneumonia requiring hospitalization. AB - From February 1995 through May 1997, we prospectively studied 533 patients with community-acquired pneumonia requiring hospitalization in order to assess the current usefulness of sputum Gram stain in guiding the etiologic diagnosis and initial antibiotic therapy when applied routinely. Sputum samples of good quality were obtained in 210 (39%) patients, 175 of whom showed a predominant morphotype. Sensitivity and specificity of Gram stain for the diagnosis of pneumococcal pneumonia were 57% and 97%, respectively; the corresponding values for Haemophilus influenzae pneumonia were 82% and 99%. Patients with a predominant morphotype were more frequently treated with monotherapy than were patients without a demonstrative sputum sample (89% vs. 75%; P<.001). Analysis of our data shows that a good-quality sputum sample can be obtained from a substantial number of patients with community-acquired pneumonia. Gram stain was highly specific for the diagnosis of pneumococcal and H. influenzae pneumonia and may be useful in guiding pathogen-oriented antimicrobial therapy. PMID- 11049764 TI - Effects of amoxicillin/clavulanate or azithromycin on nasopharyngeal carriage of Streptococcus pneumoniae and Haemophilus influenzae in children with acute otitis media. AB - The effect of antibiotic therapy on nasopharyngeal colonization by Streptococcus pneumoniae and Haemophilus influenzae was evaluated in children diagnosed with acute otitis media. Children were randomly assigned to receive either amoxicillin/clavulanate or azithromycin therapy, and nasopharyngeal swabs were obtained for culture before and after starting therapy. Amoxicillin/clavulanate therapy eradicated or suppressed all strains of S. pneumoniae susceptible to penicillin, 75% of strains with intermediate resistance, and 40% of strains resistant to penicillin. Azithromycin therapy cleared two-thirds of azithromycin susceptible strains of S. pneumoniae but none of azithromycin-nonsusceptible strains. Selection for antibiotic-resistant strains in individual children was not observed in children who received amoxicillin/clavulanate therapy but was observed in 2 children who received azithromycin therapy. Carriage of H. influenzae was also reduced by antimicrobial therapy but more so by amoxicillin/clavulanate. Antibiotic therapy does not directly increase the number of resistant strains in the population but, by eradicating susceptible strains, allows greater opportunity for carriage and spread of resistant strains. PMID- 11049765 TI - Human herpesvirus 6 infection after autologous or allogeneic stem cell transplantation: a single-center prospective longitudinal study of 92 patients. AB - To determine the incidence and clinical relevance of active human herpesvirus 6 (HHV-6) infection, 92 consecutive unselected recipients of autologous or allogeneic stem cell grafts were investigated in a prospective longitudinal study. Active infection was assessed by the presence of viral deoxyribonucleic acid (DNA) in 846 peripheral blood mononuclear cell specimens and 115 plasma specimens, by means of a specially developed polymerase chain reaction designed to avoid detection of latent genome. The incidence of HHV-6 infection observed was 42.5%, irrespective of the type or source of graft, and infection was significantly associated with partial (P=.002) or total myelosuppression (P=.01) and fever (P<. 000001). Infusion of bone marrow as the source of graft, reactivation occurring before platelet or neutrophil engraftment, and presence of HHV-6 DNA in plasma were identified as risk factors for symptomatic HHV-6 infection (P<.002). PMID- 11049766 TI - Antimicrobial use and colonization with erythromycin-resistant Streptococcus pneumoniae in Greece during the first 2 years of life. AB - We evaluated nasopharyngeal colonization with erythromycin-resistant Streptococcus pneumoniae during the first 2 years of life in central and southern Greece. Of 2448 children studied from February 1997 to February 1999, 766 (31%) carried 781 pneumococcal isolates. Ninety-five (3.9%) of the children attended day care centers. Eighteen percent of the pneumococci were resistant to erythromycin (minimal inhibitory concentration 1 to >128 microg/mL), with 67.9% of them carrying the erm(B) gene and 29.2% mef(A) gene products. Four strains possessed neither the erm(B) nor the mef(A) gene. Multidrug resistance occurred in 97% and 40% of isolates carrying the erm(B) and mef(A) gene, respectively. An association was found between the erm(B) gene and serotypes 6B and 23F and between the mef(A) gene and serotypes 14 and 19F. A significant relationship existed between carriage of erythromycin-resistant pneumococci and use of macrolides or beta-lactams in the previous 3 months; the association was strongest when macrolide therapy was administered during the last month (odds ratio, 5.92; P=.0001). The findings indicate the necessity of a judicious use of both macrolides and beta-lactams in young children to reduce the colonization with erythromycin-resistant pneumococci and the subsequent spread of such strains to the community. PMID- 11049767 TI - Human herpesvirus 6 and multiple sclerosis: systemic active infections in patients with early disease. AB - By means of immunohistochemical staining, cells actively infected with human herpesvirus 6 (HHV-6) were found in central nervous system tissues from 8 (73%) of 11 patients with definite multiple sclerosis (MS). Interestingly, 17 (90%) of 19 tissue sections showing active demyelination were positive for HHV-6-infected cells compared with only 3 (13%) of 23 tissue sections free of active disease (P<.0001). Central nervous system tissues from 2 of 28 normal persons and patients with other inflammatory demyelinative diseases were positive for HHV-6 infected cells (P<.0001), and the 2 positive cases were diagnosed as having HHV-6 leukoencephalitis. By use of a rapid culture assay, blood samples from 22 (54%) of 41 patients with definite MS were found to contain active HHV-6 infections, compared with 0 of 61 normal controls (P<.0001). No significant difference was found between HHV-6 viremia-positive and HHV-6 viremia-negative MS patients with respect to type of disease (relapsing/remitting or progressive). In contrast, patients with active HHV-6 viremia were significantly younger and had shorter durations of disease than did HHV-6 viremia-negative patients. PMID- 11049768 TI - Thrombocytopenic purpura associated with brucellosis: report of 2 cases and literature review. AB - Mild hematologic abnormalities are common in the course of human brucellosis; however, they generally resolve promptly with treatment of the disease. Occasionally, thrombocytopenia is severe and can be associated with bleeding into the skin (purpura) and from mucosal sites. We describe 2 patients infected with Brucella melitensis who presented with thrombocytopenic purpura, and we review 41 additional cases from the literature. Patients ranged in age from 2 to 77 years, and both sexes were affected equally. In the majority of cases, examination of the bone marrow revealed abundant megakaryocytes. Possible mechanisms involved in thrombocytopenia include hypersplenism, reactive hemophagocytosis, and immune destruction of platelets. Recognition of this complication is essential, since hemorrhage into the central nervous system is associated with a high mortality rate. PMID- 11049769 TI - Cluster of pulmonary infections caused by Cunninghamella bertholletiae in immunocompromised patients. AB - Cunninghamella bertholletiae is a rare cause of pulmonary mucormycosis. We describe a cluster of invasive pulmonary infections caused by C. bertholletiae in 4 immunocompromised patients that occurred during a 2-year period at 1 center. Three of the patients were receiving antifungal prophylaxis with itraconazole. Presenting symptoms were fever unresponsive to antibacterial chemotherapy, hemoptysis, and infiltrates on chest radiograms. Three patients were treated with liposomal amphotericin B. Only 1 patient survived. PMID- 11049770 TI - Rapid diagnosis of infectious pleural effusions by use of reagent strips. AB - Reagent strips have not yet been tested for use in the diagnosis of infectious pleural effusions. A reagent strip was used to evaluate 82 patients with pleural effusions: 20 patients had transudative effusions, 35 had infectious exudative effusions (empyema in 14 and parapneumonic effusion in 21), and 27 had noninfectious exudative effusions. Pleural fluid protein, as evaluated by the reagent strip, proved accurate for the detection of exudative effusions (sensitivity, 93.1%; specificity, 50%; positive predictive value, 84.3%; negative predictive value, 71.5%; odds ratio [OR], 6.77; and 95% confidence interval [CI], 1.87-24). The reagent strip leukocyte esterase test effectively detected infectious exudative effusions (sensitivity, 42.8%; specificity, 91.3%; positive predictive value, 88.2%; negative predictive value, 51.2%; OR, 4.46; and 95% CI, 1.2-16.4). Pleural pH was significantly predicted by the reagent strip but was of no assistance in categorization of exudative effusions as infectious or noninfectious. Compared with physical, laboratory, and microbiological data, the reagent strip was as accurate for estimation of percentages of infectious and noninfectious exudative effusions. Thus, reagent strips may be a rapid, easy-to use, and inexpensive technique for discriminating transudative from exudative pleural effusions and for categorizing exudative pleural effusions as infectious or noninfectious. PMID- 11049771 TI - Prevalence of sexually transmitted infections and associated risk factors among populations of drug abusers. AB - A cross-sectional survey was conducted of sexually transmitted diseases (STDs) and risky behaviors among 407 drug abusers in treatment facilities in 1998. Infections with human immunodeficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV), herpes simplex virus type 2 (HSV-2), and syphilis were detected by testing serum antibody levels; chlamydia and gonorrhea were detected by testing nucleic acid levels in urine. Logistic regression analysis was performed to measure associations. Prevalences of antibodies were as follows: to HSV-2, 44.4%; to HCV, 35.1%; to HBV, 29.5%; to HIV, 2.7%. The prevalence of syphilis was 3.4%; of chlamydia, 3.7%; and of gonorrhea, 1.7%. Of the 407 subjects, approximately 62% had markers for 1 of the STDs. HIV infection was associated with African American race, use of smokable freebase (crack) cocaine, and STD history. HBV infection was associated with age >30 years, injecting drugs, needle sharing, a history of treatment for drug abuse, and African American race. HCV infection was associated with an age >30 years, injecting drugs, and needle sharing, and HSV-2 infection with an age >30 years, female sex, and African American race. Syphilis was associated with a history of STDs. High prevalences of STDs among drug abusers indicate the need for integration of STD screening and treatment into drug treatment programs. PMID- 11049772 TI - Resistant herpes simplex virus type 1 infection: an emerging concern after allogeneic stem cell transplantation. AB - Fourteen cases of severe acyclovir-resistant herpes simplex virus type 1 (HSV-1) infection, 7 of which showed resistance to foscarnet, were diagnosed among 196 allogeneic stem cell transplant recipients within a 29-month period. Recipients of unrelated stem cell transplants were at higher risk. All patients received foscarnet; 8 subsequently received cidofovir. Strains were initially foscarnet resistant in 3 patients and secondarily so in 4 patients. In vitro resistance to acyclovir or foscarnet was associated with clinical failure of these drugs; however, in vitro susceptibility to foscarnet was associated with complete response in only 5 of 7 patients. No strain from any of the 7 patients was resistant in vitro to cidofovir; however, only 3 of 7 patients achieved complete response. Therefore, acyclovir- and/or foscarnet-resistant HSV-1 infections after allogeneic stem cell transplantation have become a concern; current strategies need to be reassessed and new strategies must be evaluated in this setting. PMID- 11049773 TI - First European exposure to syphilis: the Dominican Republic at the time of Columbian contact. AB - Recognition of syphilis in Europe in the late 15th century and its prior absence suggest New World origin. Skeletal populations were examined from sites with documented Columbian contact in the Dominican Republic. Examination of 536 skeletal remains revealed periosteal reaction characteristic of treponemal disease in 6%-14% of the afflicted population. Findings were identical to that previously noted in confirmed syphilis-affected populations and distinctive from those associated with yaws and bejel: it was a low population frequency phenomenon, affecting an average of 1.7-2.6 bone groups, often asymmetric and sparing hands and feet, but associated with significant tibial remodeling. While findings diagnostic of syphilis have been reported in the New World, actual demonstration of syphilis in areas where Columbus actually had contact was missing, until now. The evidence is consistent with this site as the point of initial contact of syphilis and of its subsequent spread from the New World to the Old. PMID- 11049774 TI - Hantavirus pulmonary syndrome in northern Alberta, Canada: clinical and laboratory findings for 19 cases. AB - We reviewed the clinical and laboratory findings for 19 cases of hantavirus pulmonary syndrome (HPS) identified either serologically or by immunohistochemical testing of archival tissue at our tertiary care center. Fever (95%), cough (89%), and dyspnea (89%) were the most common presenting symptoms. The most prevalent presenting signs were respiratory abnormalities (95%) and tachycardia (84%). Common laboratory findings included thrombocytopenia (95%) and leukocytosis (79%). Elevated aspartate aminotransferase and lactate dehydrogenase levels were found in all patients tested. Intubation was required in 58% of the patients, and inotropic support was required in 53%. Our study confirms that serological responses appear early during clinical illness, making the enzyme immunoassay a useful tool for the diagnosis of acute HPS. The mortality (26%) and severity of disease that we observed among patients with HPS appear to be less than those reported elsewhere. PMID- 11049775 TI - Risk of Cryptosporidium parvum transmission between hospital roommates. AB - Patients with active diarrhea caused by infection with Cryptosporidium parvum can potentially contaminate the environment, which could serve as a risk for transmission to other patients in a hospital setting. A retrospective cohort study was performed to quantify the risk of nosocomial roommate-to-roommate transmission of Cryptosporidium and to evaluate the need for isolation of Cryptosporidium-infected patients. Thirty-seven human immunodeficiency virus (HIV)-infected roommates of 21 index patients with Cryptosporidium were identified between 1994 and 1996. Each exposed roommate (median CD4 cell count, 27cells/mm(3)) was matched to an HIV-infected, unexposed roommate with a similar CD4 cell count (median, 24 cells/mm(3)) who was present in the hospital during the same month but was not a roommate of a patient with Cryptosporidium infection. No patients with Cryptosporidium were identified among the 37 exposed roommates, and 1 case was identified among the 37 unexposed roommates. The risk ratio for chronic diarrhea was 0.80 (95% confidence interval [CI], 0.23-2.75) and for death was 1.04 (95% CI, 0.75-1.44). These results suggest that isolation of adult patients with Cryptosporidium diarrhea is not necessary to prevent roommate to-roommate transmission of Cryptosporidium. PMID- 11049776 TI - Home sampling versus conventional swab sampling for screening of Chlamydia trachomatis in women: a cluster-randomized 1-year follow-up study. AB - We compared the efficacy of a screening program for urogenital Chlamydia trachomatis infections based on home sampling with that of a screening program based on conventional swab sampling performed at a physician's office. Female subjects, comprising students at 17 high schools in the county of Aarhus, Denmark, were divided into a study group (tested by home sampling) and a control group (tested in a physician's office). We assessed the number of new infections and the number of subjects who reported being treated for pelvic inflammatory disease (PID) at 1 year of follow-up; 443 (51.1%) of 867 women in the intervention group and 487 (58.5%) of 833 women in the control group were available for follow-up. Thirteen (2.9%) and 32 (6.6%) new infections were identified in the intervention group and the control group, respectively (Wilcoxon exact value, P=.026). Nine (2.1%) women in the intervention group and 20 (4.2%) in the control group reported being treated for PID (P=.045), indicating that a screening strategy involving home sampling is associated with a lower prevalence of C. trachomatis and a lower proportion of reported cases of PID. PMID- 11049777 TI - Nontuberculous mycobacterial disease in northern Australia: a case series and review of the literature. AB - We performed a retrospective/prospective review of all cases of disease due to nontuberculous mycobacteria (NTM) reported in the Northern Territory, Australia, during the period 1989-1997. Fifty-eight cases were reported, with an average yearly incidence of 3.9 cases per 100,000 persons. The number increased significantly for the second half of the study period (39 vs. 19 cases; P<.02). The yearly incidence of pulmonary Mycobacterium avium/Mycobacterium intracellulare complex (MAC) disease not associated with human immunodeficiency virus (HIV) infection was 2.1 cases per 100,000 population. MAC was the most common isolate (78%) and pulmonary disease the most frequent clinical presentation (62%). Disease due to NTM or MAC was not found more commonly in rural areas. Significant risks for non-HIV-associated pulmonary MAC disease included male sex (odds ratio [OR], 2.1; 95% confidence interval [CI], 1.0-4.5) and age >50 years (OR, 26.5; 95% CI, 10.9-67.3), but aboriginal people appeared underrepresented (OR, 0.77; 95% CI, 0.30-1.87). Mycobacterium tuberculosis was almost 5 times more likely than NTM to be the cause of non-HIV-associated mycobacterial pulmonary disease (153 vs. 32 cases; OR, 4.79; 95% CI, 3.22-7.14). Mycobacterial lymphadenitis in aboriginal children was more likely to be tuberculous than nontuberculous (OR, 6.5; 95% CI, 1.4-41.7), but not in nonaboriginal children (OR, 1.0). With treatment, 66% of the cases of non-HIV associated pulmonary MAC disease had favorable outcomes, and 7% of patients had progressive fatal disease. Outcomes of therapy for lymphadenitis and skin/soft tissue disease were excellent, but those of HIV-associated disseminated MAC disease were poor. PMID- 11049778 TI - Nocardia infection in heart-lung transplant recipients at Alfred Hospital, Melbourne, Australia, 1989-1998. AB - Nocardia infections are uncommon in recipients of heart, lung, or heart-lung transplants, but such infections are well described. Frequent episodes of rejection, high-dose prednisolone treatment, renal impairment, and prolonged respiratory support have all been shown to increase the risk of Nocardia infection in this group. In this retrospective review of 540 recipients of heart, lung, or heart-lung transplants, 10 patients developed Nocardia infection (frequency, 1.85%). Infection occurred at a mean +/- standard deviation of 13+/ 14.5 months after transplantation. All patients had pulmonary disease with no evidence of extrapulmonary disease. The Nocardia infection did not contribute directly to patient deaths. Coinfection with other pathogens was present in 6 patients, and 2 patients had sequential infections. Radiological findings varied. All isolates were susceptible to trimethoprim-sulfamethoxazole, amikacin, and imipenem. Treatment regimens varied. Two (30%) of 6 patients treated with trimethoprim-sulfamethoxazole developed adverse reactions, which necessitated a change in antibiotic therapy. The optimal treatment regimen, which comprises both the antimicrobial agent and the length of treatment, is unclear. PMID- 11049779 TI - Clinical and immunological risk factors associated with Haemophilus influenzae type b conjugate vaccine failure in childhood. AB - Haemophilus influenzae type b (Hib) conjugate vaccines have proved extremely efficacious in healthy children. True Hib vaccine failures are rare. Hib conjugate vaccines were introduced for routine immunization in the United Kingdom and the Republic of Ireland in 1992. Coincident with this, active prospective and national surveillance via pediatricians, microbiologists, and public health physicians was commenced to assess the clinical and immunological factors associated with vaccine failure. During the 6 years of the study, 115 children with true vaccine failure were reported. Of the children who were vaccinated before 12 months of age, a clinical risk factor was detected in 20%, an immunological deficiency was detected in 30%, and one or both were detected in 44%. Children who were vaccinated after 12 months of age were more likely to have one or both factors (67%). Thirty percent (33 of 105) of children with true vaccine failure had a low Hib antibody response (concentration, <1.0 microg/mL) after disease, but the majority then responded to a further dose of Hib vaccine. Children who develop Hib disease despite vaccination deserve further clinical and immunological evaluation. PMID- 11049780 TI - Endemic melioidosis in tropical northern Australia: a 10-year prospective study and review of the literature. AB - In a prospective study of melioidosis in northern Australia, 252 cases were found over 10 years. Of these, 46% were bacteremic, and 49 (19%) patients died. Despite administration of ceftazidime or carbapenems, mortality was 86% (43 of 50 patients) among those with septic shock. Pneumonia accounted for 127 presentations (50%) and genitourinary infections for 37 (15%), with 35 men (18%) having prostatic abscesses. Other presentations included skin abscesses (32 patients; 13%), osteomyelitis and/or septic arthritis (9; 4%), soft tissue abscesses (10; 4%), and encephalomyelitis (10; 4%). Risk factors included diabetes (37%), excessive alcohol intake (39%), chronic lung disease (27%), chronic renal disease (10%), and consumption of kava (8%). Only 1 death occurred among the 51 patients (20%) with no risk factors (relative risk, 0.08; 95% confidence interval, 0.01-0.58). Intensive therapy with ceftazidime or carbapenems, followed by at least 3 months of eradication therapy with trimethoprim-sulfamethoxazole, was associated with decreased mortality. Strategies are needed to decrease the high mortality with melioidosis septic shock. Preliminary data on granulocyte colony-stimulating factor therapy are very encouraging. PMID- 11049781 TI - Long-term outcome and treatment modifications in a prospective cohort of human immunodeficiency virus type 1-infected patients on triple-drug antiretroviral regimens. Triest Cohort Investigators. AB - We designed a cohort in order to assess the long-term effects of triple-drug antiretroviral combinations in 608 patients infected with human immunodeficiency virus type 1 (HIV-1). We recruited patients who had been previously treated with nucleoside analogues as well as treatment-naive patients who were starting triple drug antiretroviral combinations consisting of nucleoside analogues, either alone or in combination with a protease inhibitor. After a median follow-up time of 22 months, the incidence rates of acquired immune deficiency syndrome-defining events and death were, respectively, 6.9 (95% confidence interval [CI], 5.3-8.8) and 2.9 (95% CI, 1.9-4.2) per 100 person-years. Advanced clinical stage of disease (P=.004), a low CD4(+) cell count (P=.002), and a low quality-of-life score (P=.001) at baseline were independent predictors of clinical progression. The initial triple-drug combination was modified a total of 647 times in 321 patients. The only independent predictor of treatment modification was previous exposure to a nucleoside analogue in patients who did not receive a new nucleoside analogue at inclusion (P=.001). Plasma HIV RNA values below 500 copies/mL were obtained in 88% of the treatment-naive patients and in 57% of the previously treated patients (P<.001). Compared with previously treated patients who received > or = 1 new nucleoside analogue at enrollment, previously treated patients who did not receive a new nucleoside analogue at enrollment were twice as likely to have plasma HIV RNA values >500 copies/mL at the last visit (adjusted odds ratio [OR], 1.8; 95% confidence interval [CI], 1.2-2.8), and the antiretroviral-naive patients were significantly less likely to have plasma HIV RNA values >500 copies/mL at the last visit (adjusted OR, 0.2; 95% CI, 0.1-0.4). PMID- 11049782 TI - Environmental control to reduce transmission of Clostridium difficile. AB - Restrictive antibiotic policies and infection control measures have been shown to reduce the incidence of Clostridium difficile-associated diarrhea (CDAD) among hospitalized patients. To date, the role of environmental disinfectants in reducing nosocomial CDAD rates has not been well studied. In a before-and-after intervention study, patients in 3 units were evaluated to determine if unbuffered 1:10 hypochlorite solution is effective as an environmental disinfectant in reducing the incidence of CDAD. Among 4252 patients, the incidence rate of CDAD for bone marrow transplant patients decreased significantly, from 8.6 to 3.3 cases per 1000 patient-days (hazard ratio, 0.37; 95% confidence interval, 0.19 0.74), after the environmental disinfectant was switched from quaternary ammonium to 1:10 hypochlorite solution in the rooms of patients with CDAD. Reverting later to quaternary ammonium solution increased the CDAD rate to 8.1 cases per 1000 patient-days. No reduction in CDAD rates was seen among neurosurgical intensive care unit and general medicine patients, for whom baseline rates were 3.0 and 1.3 cases per 1000 patient-days, respectively. Unbuffered 1:10 hypochlorite solution is effective in decreasing patients' risk of developing CDAD in areas where CDAD is highly endemic. Presumed mechanisms include reducing the environmental burden and the potential for C. difficile transmission among susceptible patients. PMID- 11049783 TI - Prospective randomized trial of 10% povidone-iodine versus 0.5% tincture of chlorhexidine as cutaneous antisepsis for prevention of central venous catheter infection. AB - A multicenter prospective, randomized, controlled trial, with 0.5% tincture of chlorhexidene versus 10% povidone-iodine as cutaneous antisepsis for central venous catheter (CVC) insertion, was conducted for patients in intensive care units. Of 374 patients, 242 had a CVC inserted for >3 days and were used for the primary analysis. Outcomes included catheter-related bacteremia, significant catheter colonization (> or = 15 colony-forming units [cfu]), exit-site infection, serial quantitative exit-site culture (every 72 h), and molecular subtyping of all isolates. Patients in both study groups were comparable with respect to age, sex, underlying disease, length of hospitalization, reason for line insertion, and baseline APACHE II score. Documented catheter-related bacteremia rates were 4.6 cases per 1000 catheter-days in the chlorhexidine group (n=125) and 4.1 cases per 1000 catheter-days in the povidone-iodine group (n=117; not significant [NS]). Significant catheter-tip colonization occurred in 24 (27%) of 88 patients in the povidone-iodine group and in 31 (34%) of 92 patients in the chlorhexidine group (NS). A mean exit-site colony count of 5.9 x 10(5) cfu/mL per 25 cm(2) of the surface area of skin in the povidone-iodine group versus 3.1 x 10(5) cfu/mL per 25 cm(2) in the chlorhexidine group (NS) was found. There was a trend toward fewer exit-site infections in the chlorhexidine group (0 of 125 patients) versus those in the povidone-iodine group (4 of 117 patients; P=.053). Results of an intention-to-treat analysis were unchanged from the primary analysis. No difference was demonstrable between 0.5% tincture of chlorhexidine and 10% povidone-iodine when used for cutaneous antisepsis for CVC insertion in patients in the intensive care unit. PMID- 11049784 TI - Breakthrough pneumococcal bacteremia in patients being treated with azithromycin and clarithromycin. AB - Streptococcus pneumoniae strains have exhibited decreasing susceptibility to penicillins and macrolides during the past several years. We reviewed the medical charts of all patients with pneumococcal bacteremia who were admitted to a university hospital over a period of 1 year, to identify failures of outpatient therapy. Of 41 patients admitted with pneumococcal bacteremia, 4 had previously taken either azithromycin or clarithromycin for 3-5 days. All 4 had pneumococcal strains that exhibited low-level resistance to macrolide antibiotics. Among pneumococci, low-level resistance to macrolides can lead to clinical failure, and resistance to macrolides should be considered during the selection of empiric therapy for patients with presumed pneumococcal infections. PMID- 11049785 TI - The search for a better treatment for recurrent Clostridium difficile disease: use of high-dose vancomycin combined with Saccharomyces boulardii. AB - Recurrent Clostridium difficile disease (CDD) is a difficult clinical problem because antibiotic therapy often does not prevent further recurrences. In a previous study, the biotherapeutic agent Saccharomyces boulardii was used in combination with standard antibiotics and was found to be effective in reducing subsequent recurrences of CDD. In an effort to further refine a standard regimen, we tested patients receiving a regimen of a standard antibiotic for 10 days and then added either S. boulardii (1 g/day for 28 days) or placebo. A significant decrease in recurrences was observed only in patients treated with high-dose vancomycin (2 g/day) and S. boulardii (16.7%), compared with those who received high-dose vancomycin and placebo (50%; P=.05). No serious adverse reactions were observed in these patients. Comparison of data from this trial with data from previous studies indicates that recurrent CDD may respond to a short course of high-dose vancomycin or to longer courses of low-dose vancomycin when either is combined with S. boulardii. PMID- 11049786 TI - Wound botulism in California, 1951-1998: recent epidemic in heroin injectors. AB - California has reported most of the world's wound botulism (WB) cases and nearly three-fourths of the cases reported in the United States. We reviewed the clinical, epidemiologic, and laboratory features of WB. From the first case in 1951, through 1998, a total of 127 cases were identified-93 in the last 5 years. The dramatic increase has been due to an epidemic (of WB) in people who inject black tar heroin. Whereas early cases of WB occurred after gross trauma, all but 1 of the last 102 cases occurred in drug users, primarily those who inject drugs subcutaneously ("skin poppers"). Cases are occurring disproportionately in Hispanics and women. Misdiagnosis and diagnostic delays of up to 64 days have occurred. This unprecedented, ongoing epidemic is now being reported in other states. We discuss the clinical and laboratory features that distinguish botulism from conditions that can mimic it, the relative yield of various diagnostic laboratory tests for botulism, and its treatment. PMID- 11049787 TI - Acute onset of type I diabetes mellitus after severe echovirus 9 infection: putative pathogenic pathways. AB - Enterovirus infections have been implicated in the development of type I diabetes mellitus. They may cause beta cell destruction either by cytolytic infection in the pancreas or indirectly by contributing to autoimmune reactivity. We sought evidence for these 2 mechanisms in a case of acute-onset diabetes mellitus that occurred during severe echovirus 9 infection. The virus was isolated and administered to cultured human beta cells. No viral proliferation was observed, and no beta cell death was induced, while parallel exposure to Coxsackie B virus serotype 3 resulted in viral proliferation and massive beta cell death. Although the viral protein 2C exhibited a sequence similar to that of the beta cell autoantigen glutamic acid decarboxylase (GAD(65)), no cross-reactive T cell responses were detected. The patient did not develop antibodies to GAD(65) either. Absence of evidence for direct cytolytic action or an indirect effect through molecular mimicry with GAD(65) in the present case raises the possibility of another indirect pathway through which enteroviruses can cause diabetes mellitus. PMID- 11049788 TI - Paracoccidioidomycosis: a model for evaluation of the effects of human immunodeficiency virus infection on the natural history of endemic tropical diseases. AB - The interaction of human immunodeficiency virus (HIV) infection with endemic tropical diseases has become a major concern, but its mechanisms are still poorly understood. Paracoccidioidomycosis (PCM), a South America endemic deep mycosis, may provide an interesting model to investigate this interaction, as clinical epidemiological features of most HIV-PCM-coinfected patients are difficult to classify into the standard acute and chronic forms of PCM. Such patients have presented clinical features indicative of an uncontrolled infection with lymphohematogenous dissemination, similar to the more severe, acute form. However, this infection probably resulted from reactivated latent foci that, in nonimmunocompromised hosts, leads to the less severe chronic form, characterized by mucosal lesions. We propose that a new outcome of the Paracoccidioides brasiliensis-host interaction is induced by concomitant HIV infection. This outcome probably reflects an impaired anti-P. brasiliensis immune response during coinfection that is similar to that seen in the acute form, although the patients have a chronic P. brasiliensis infection. PMID- 11049789 TI - Cyclospora cayetanensis: a review, focusing on the outbreaks of cyclosporiasis in the 1990s. AB - Cyclospora cayetanensis, a coccidian parasite that causes protracted, relapsing gastroenteritis, has a short recorded history. In retrospect, the first 3 documented human cases of Cyclospora infection were diagnosed in 1977 and 1978. However, not much was published about the organism until the 1990s. One of the surprises has been the fact that a parasite that likely requires days to weeks outside the host to become infectious has repeatedly caused foodborne outbreaks, including large multistate outbreaks in the United States and Canada. In this review, I discuss what has been learned about this enigmatic parasite since its discovery and what some of the remaining questions are. My focus is the foodborne and waterborne outbreaks of cyclosporiasis that were documented from 1990 through 1999. The occurrence of the outbreaks highlights the need for health care personnel to consider that seemingly isolated cases of infection could be part of widespread outbreaks and should be reported to public health officials. Health care personnel should also be aware that stool specimens examined for ova and parasites usually are not examined for Cyclospora unless such testing is specifically requested and that Cyclospora infection is treatable with trimethoprim-sulfamethoxazole. PMID- 11049790 TI - Insights into the epidemiology and control of infection with vancomycin-resistant enterococci. AB - Despite control efforts, the incidence of nosocomial infections due to vancomycin resistant enterococci (VRE) continues to increase in the United States. VRE are thought to spread primarily by cross-contamination. Recent molecular epidemiologic studies have refined our understanding of this phenomenon. If VRE are not controlled soon after introduction into a hospital, sporadic cases may evolve into a monoclonal outbreak, which may then evolve to polyclonal endemicity. An intervention that is effective in containing VRE in one setting may be ineffective in another. Control of VRE where they are endemic is particularly challenging. Although eradication of endemic VRE may not be possible, aggressive, multifaceted programs have been successful in diminishing the problem. A mathematical model of transmission of VRE and the effect of infection control measures in settings where they are endemic has been reported. The use of such a model may allow more precise determination of the impact of control strategies in the future. PMID- 11049791 TI - Community-acquired pneumonia in the elderly. AB - Pneumonia in the elderly is a common and serious problem with a clinical presentation that can differ from that in younger patients. Older patients with pneumonia complain of significantly fewer symptoms than do younger patients, and delirium commonly occurs. Indeed, delirium may be the only manifestation of pneumonia in this group of patients. Alcoholism, asthma, immunosuppression, and age >70 years are risk factors for community-acquired pneumonia in the elderly. Among nursing home residents, the following are risk factors for pneumonia: advanced age, male sex, difficulty in swallowing, inability to take oral medications, profound disability, bedridden state, and urinary incontinence. Streptococcus pneumoniae is the most common cause of pneumonia among the elderly. Aspiration pneumonia is underdiagnosed in this group of patients, and tuberculosis always should be considered. In this population an etiologic diagnosis is rarely available when antimicrobial therapy must be instituted. Use of the guidelines for treatment of pneumonia issued by the Infectious Diseases Society of America, with modification for treatment in the nursing home setting, is recommended. PMID- 11049792 TI - Empirical antimicrobial therapy for traveler's diarrhea. AB - Over 7 million cases of traveler's diarrhea, defined as the passage of > or = 3 unformed stools in a 24-h period, occur each year among visitors to developing countries. Bacterial enteric pathogens are the most common etiologic agents isolated. Preliminary clinical results for patients with diarrhea predominantly caused by Campylobacter species have shown that azithromycin may be an effective alternative to fluoroquinolones for the treatment of traveler's diarrhea. PMID- 11049793 TI - Paromomycin: no more effective than placebo for treatment of cryptosporidiosis in patients with advanced human immunodeficiency virus infection. AIDS Clinical Trial Group. AB - To evaluate the efficacy of paromomycin for the treatment of symptomatic cryptosporidial enteritis in human immunodeficiency virus-infected adults, we conducted a prospective, randomized, double-blind, placebo-controlled trial before the widespread introduction of highly active antiretroviral therapy (HAART). Seven units under the auspices of the AIDS Clinical Trials Group enrolled 35 adults with CD4 cell counts of < or = 150/mm(3). Initially, 17 patients received paromomycin (500 mg 4 times daily) and 18 received matching placebo for 21 days. Then all patients received paromomycin (500 mg q.i.d.) for an additional 21 days. Clinical definitions of response were measured by an average number of bowel movements per day in association with concurrent need for antidiarrheal agents that was lower than that before study entry. There was no treatment response during the placebo-controlled phase of the study according to protocol-defined criteria (P=.88). Three paromomycin recipients (17.6%) versus 2 placebo recipients (14.3%) responded completely. Rates of combined partial and complete responses in the paromomycin arm (8 out of 17, 47.1%) and the placebo arm (5 out of 14, 35.7%) of the study were also similar (P=.72). The clinical course of cryptosporidiosis was quite variable. Paromomycin was not shown to be more effective than placebo for the treatment of symptomatic cryptosporidial enteritis. However, inadequate statistical power prevents definitive rejection of the usefulness of paromomycin as therapy for this infection. PMID- 11049794 TI - HIV and Leishmania coinfection: a review of 91 cases with focus on atypical locations of Leishmania. AB - A retrospective study was conducted in France in 1998 to determine the clinical features of visceral leishmaniasis (VL) in 91 patients infected cocomitantly with human immunodeficiency virus. Our data suggest that the clinical manifestations of VL may be influenced by the immunological status, with atypical locations of Leishmania amastigotes more frequently found in severely immunocompromised patients. In such patients, the involvement of atypical locations may lead to the discovery of VL. PMID- 11049795 TI - Prolonged suppression of human immunodeficiency virus type 1 RNA during dual nucleoside reverse-transcriptase-inhibitor therapy in clinical practice. AB - Because there is limited information about suppression of virus loads (determined by current "ultrasensitive" assays) in patients receiving nucleoside reverse transcriptase inhibitors (NRTIs) alone, we reviewed our experience in clinical practice with patients who had virus loads of <25 copies/mL after >1 year of treatment with dual NRTIs. PMID- 11049796 TI - Human immunodeficiency virus type 1 infection after sexual abuse: value of nucleic acid sequence analysis in identifying the offender. AB - Nucleic acid sequence analysis of human immunodeficiency virus type 1-specific sequences allowed the identification of the source of infection in a case of sexual abuse of a 10-year-old girl. PMID- 11049797 TI - Outbreak of amebiasis in a family in The Netherlands. AB - Human-to-human transmission of Entamoeba histolytica is rare in industrialized countries. We describe an outbreak of amebiasis in a family in The Netherlands, demonstrating that even with Western standards of hygiene, persistent cyst passage may result in the transmission of E. histolytica to household contacts. If E. histolytica is isolated from a person living in an area of nonendemicity, it may be worthwhile to test all family members for cyst passage. PMID- 11049798 TI - Failure of pentavalent antimony in visceral leishmaniasis in India: report from the center of the Indian epidemic. AB - In India, 320 patients with visceral leishmaniasis (209 in the state of Bihar and 11 in the neighboring state of Uttar Pradesh) received identical pentavalent antimony (Sb) treatment. Sb induced long-term cure in 35% (95% confidence interval [CI], 28%-42%) of those in Bihar versus 86% (95% CI, 79%-93%) of those in Uttar Pradesh. In Bihar, the center of the Indian epidemic, traditional Sb treatment should be abandoned. PMID- 11049799 TI - Death from inappropriate therapy for Lyme disease. AB - A 30-year-old woman died as a result of a large Candida parapsilosis septic thrombus located on the tip of a Groshong catheter. The catheter had been in place for 28 months for administration of a 27 month course of intravenous cefotaxime for an unsubstantiated diagnosis of chronic Lyme disease. PMID- 11049800 TI - Short-course of oral miltefosine for treatment of visceral leishmaniasis. AB - A total of 54 Indian patients with visceral leishmaniasis were treated with oral miltefosine, 50 mg given twice daily, for 14 days (18 patients; group A), 21 days (18; group B), or 28 days (18; group C). Cure was achieved in 89% of group A, 100% of group B, and 100% of group C. Adverse reactions were self-limited and primarily mild. The 21-day miltefosine regimen combines high-level efficacy, convenient dosing, and a relatively short duration. PMID- 11049801 TI - La Crosse encephalitis presenting like herpes simplex encephalitis in an immunocompromised adult. AB - The diagnosis of the precise cause of viral encephalitis can be difficult, hampered by the nonspecific presentation, the number of etiologic viruses, and limited culture and serologic diagnostic methods. Because herpes simplex encephalitis (HSE) can be neurologically devastating and is treatable, timely diagnosis is important. We report an immunocompromised adult with encephalitis clinically consistent with HSE who had serology consistent with recent La Crosse encephalitis (LAC). PMID- 11049802 TI - Profound and prolonged lymphocytopenia with West Nile encephalitis. PMID- 11049803 TI - Controlled trials of amphotericin B lipid complex. PMID- 11049804 TI - Reply PMID- 11049806 TI - Report of 2 fatal cases of adult necrotizing fasciitis and toxic shock syndrome caused by Streptococcus agalactiae. AB - We describe 2 cases of fatal necrotizing fasciitis and toxic shock syndrome caused by Streptococcus agalactiae-a rare entity that has been reported in only 9 patients-in 2 nonpregnant adults. PMID- 11049805 TI - Large outbreak in a surgical intensive care unit of colonization or infection with Pseudomonas aeruginosa that overexpressed an active efflux pump. AB - During a 30-month survey, 55 patients were colonized or infected by a single clone of Pseudomonas aeruginosa in a surgical intensive care unit (ICU). This clone overexpressed an efflux pump system, and its antibiotic resistance pattern was extremely stable as it spread from patient to patient. Pulsed-field gel electrophoresis showed that isolates from different patients were genetically identical or very similar. We were unable to identify an environmental reservoir, but cultures of hand specimens from 2 health care workers were positive. It was not clear whether this carriage was the source of the epidemic or a consequence of it. However, the propagation of the epidemic clone was probably linked to its transmission by the staff from patient to patient. The outbreak was controlled, with difficulty, by strengthening isolation procedures, replacing the antiseptic soap being used by the staff, and changing the antibiotic prescription policy. This observation emphasizes the importance of compliance with hand washing and universal precautions. PMID- 11049807 TI - Potential role of famciclovir for prevention of herpetic whitlow in the health care setting. AB - Oral famciclovir was initiated in a health care worker immediately after an accidental percutaneous injury involving a needle freshly removed from a patient's herpes labialis vesicles. In follow-up, the health care worker remained seronegative for herpes simplex I and II antibodies (IgG and IgM) and did not develop herpetic whitlow, supporting the potential role of famciclovir in the prevention of herpetic whitlow in a health care setting. PMID- 11049808 TI - Progressive multifocal leukoencephalopathy and idiopathic CD4+lymphocytopenia: a case report and review of reported cases. AB - Progressive multifocal leukoencephalopathy (PML) is a well recognized demyelinating neurological disorder caused by JC virus. Idiopathic CD4(+) lymphocytopenia (ICL) is a syndrome first described by the Centers for Disease Control and Prevention as a CD4(+) count <300 cells/mm(3) or a CD4(+) count that is <20% of the total T cell count on 2 occasions, with no evidence of human immunodeficiency virus (HIV) infection on testing, and absence of any defined immunodeficiency or therapy that depresses the levels of CD4(+) T cells. To the best of our knowledge, this is the third reported case of PML and ICL, and also the first reported case of the use of cidofovir to treat PML in a patient not infected with human immunodeficiency virus. PMID- 11049809 TI - Long-term health effects of particulate and other ambient air pollution: research can progress faster if we want it to. AB - There is need for the assessment of long-term effects of outdoor air pollution. In fact, a considerable part of the large amount of U.S. research money that has been dedicated to investigate effects of ambient particulate pollution should be invested to address long-term effects. Studies that follow the health status of large numbers of subjects across long periods of time (i.e., cohort studies) should be considered the key research approach to address these questions. However, these studies are time consuming and expensive. We propose efficient strategies to address these questions in less time. Apart from long-term continuation of the few ongoing air pollution cohort studies in the United States, data from large cohorts that were established decades ago may be efficiently used to assess cardiorespiratory effects and to target research on detection of the most susceptible subgroups in the population, which may be related to genetic, molecular, behavioral, societal, and/or environmental factors. This approach will be efficient only if the available air pollution monitoring data will be used to spatially model long-term outdoor pollution concentrations across a given country for each year with available pollution data. Such concentration maps will allow researchers to impute outdoor air pollution levels at any residential location, independent of the location of monitors. Exposure imputation may be based on residential location(s) of participants in long-standing cardiorespiratory cohort studies, which can be matched to pollutant levels using geographic information systems. As shown in European impact assessment studies, such maps may be derived relatively quickly. PMID- 11049810 TI - Human cell exposure assays of Bacillus thuringiensis commercial insecticides: production of Bacillus cereus-like cytolytic effects from outgrowth of spores. AB - Most contemporary bioinsecticides are derived from scaled-up cultures of Bacillus thuringiensis subspecies israelensis (Bti) and kurstaki (Btk), whose particulate fractions contain mostly B. thuringiensis spores (> 10(12)/L) and proteinaceous aggregates, including crystal-like parasporal inclusion bodies (PIB). Based on concerns over relatedness to B. cereus-group pathogens, we conducted extensive testing of B. thuringiensis (BT) products and their subfractions using seven human cell types. The Bti/Btk products generated nonspecific cytotoxicities involving loss in bioreduction, cell rounding, blebbing and detachment, degradation of immunodetectable proteins, and cytolysis. Their threshold dose (Dt approximately equal.5 times 10(-14)% BT product/target cell) equated to a single spore and a target cell half-life (tLD(50)) of approximately 16 hr. At Dts > 10(4), the tLD(50) rapidly shifted to < 4 hr; with antibiotic present, no component, including PIB-related [delta]-endotoxins, was cytolytic up to an equivalent of approximately 10(9 )Dt. The cytolytic agent(s) within the Bti/Btk vegetative cell exoprotein (VCP) pool is an early spore outgrowth product identical to that of B. cereus and acting possibly by arresting protein synthesis. No cytolytic effects were seen with VCP from B. subtilis and Escherichia coli. These data, including recent epidemiologic work indicate that spore-containing BT products have an inherent capacity to lyse human cells in free and interactive forms and may also act as immune sensitizers. To critically impact at the whole body level, the exposure outcome would have to be an uncontrolled infection arising from intake of Btk/Bti spores. For humans, such a condition would be rare, arising possibly in equally rare exposure scenarios involving large doses of spores and individuals with weak or impaired microbe clearance capacities and/or immune response systems. PMID- 11049811 TI - Disposition of orally administered 2,2-Bis(4-hydroxyphenyl)propane (Bisphenol A) in pregnant rats and the placental transfer to fetuses. AB - We studied the disposition of bisphenol A (BPA) in pregnant female F344/DuCrj(Fischer) rats and its placental transfer to fetuses after a single oral administration of 1 g/kg BPA dissolved in propylene glycol. BPA in maternal blood, liver, and kidney reached maximal concentrations (14.7, 171, and 36 microg/g) 20 min after the administration and gradually decreased. The levels were 2-5% of the maximum 6 hr after the administration. The maximal concentration of BPA in fetuses (9 microg/g) was also attained 20 min after the administration. BPA levels then gradually reduced in a similar manner to maternal blood. These results suggest that the absorption and distribution of BPA in maternal organs and fetuses are extremely rapid and that the placenta does not act as a barrier to BPA. PMID- 11049812 TI - Lead loading of urban streets by motor vehicle wheel weights. AB - This study documents that lead weights, which are used to balance motor vehicle wheels, are lost and deposited in urban streets, that they accumulate along the outer curb, and that they are rapidly abraded and ground into tiny pieces by vehicle traffic. The lead is so soft that half the lead deposited in the street is no longer visible after little more than 1 week. This lead loading of urban streets by motor vehicle wheel weights is continuous, significant, and widespread, and is potentially a major source of human lead exposure because the lead is concentrated along the outer curb where pedestrians are likely to step. Lead deposition at one intersection in Albuquerque, New Mexico, ranged from 50 to 70 kg/km/year (almost 11 g/ft(2)/year along the outer curb), a mass loading rate that, if accumulated for a year, would exceed federal lead hazard guidelines more than 10,000 times. Lead loading of major Albuquerque thoroughfares is estimated to be 3,730 kg/year. Wheel weight lead may be dispersed as fugitive dust, flushed periodically by storm water into nearby waterways and aquatic ecosystems, or may adhere to the shoes of pedestrians or the feet of pets, where it can be tracked into the home. I propose that lead from wheel weights contributes to the lead burden of urban populations. PMID- 11049813 TI - Association of fine particulate matter from different sources with daily mortality in six U.S. cities. AB - Previously we reported that fine particle mass (particulate matter [less than and equal to] 2.5 microm; PM(2.5)), which is primarily from combustion sources, but not coarse particle mass, which is primarily from crustal sources, was associated with daily mortality in six eastern U.S. cities (1). In this study, we used the elemental composition of size-fractionated particles to identify several distinct source-related fractions of fine particles and examined the association of these fractions with daily mortality in each of the six cities. Using specific rotation factor analysis for each city, we identified a silicon factor classified as soil and crustal material, a lead factor classified as motor vehicle exhaust, a selenium factor representing coal combustion, and up to two additional factors. We extracted daily counts of deaths from National Center for Health Statistics records and estimated city-specific associations of mortality with each source factor by Poisson regression, adjusting for time trends, weather, and the other source factors. Combined effect estimates were calculated as the inverse variance weighted mean of the city-specific estimates. In the combined analysis, a 10 microg/m(3) increase in PM(2.5) from mobile sources accounted for a 3.4% increase in daily mortality [95% confidence interval (CI), 1.7-5.2%], and the equivalent increase in fine particles from coal combustion sources accounted for a 1.1% increase [CI, 0.3-2.0%). PM(2.5) crustal particles were not associated with daily mortality. These results indicate that combustion particles in the fine fraction from mobile and coal combustion sources, but not fine crustal particles, are associated with increased mortality. PMID- 11049814 TI - Associations of blood lead, dimercaptosuccinic acid-chelatable lead, and tibia lead with polymorphisms in the vitamin D receptor and [delta]-aminolevulinic acid dehydratase genes. AB - A cross-sectional study was performed to evaluate the influence of polymorphisms in the [delta]-aminolevulinic acid dehydratase (ALAD) and vitamin D receptor (VDR) genes on blood lead, tibia lead, and dimercaptosuccinic acid (DMSA) chelatable lead levels in 798 lead workers and 135 controls without occupational lead exposure in the Republic of Korea. Tibia lead was assessed with a 30-min measurement by (109)Cd-induced K-shell X-ray fluorescence, and DMSA-chelatable lead was estimated as 4-hr urinary lead excretion after oral administration of 10 mg/kg DMSA. The primary goals of the analysis were to examine blood lead, tibia lead, and DMSA-chelatable lead levels by ALAD and VDR genotypes, controlling for covariates; and to evaluate whether ALAD and VDR genotype modified relations among the different lead biomarkers. There was a wide range of blood lead (4-86 microg/dL), tibia lead (-7-338 microg Pb/g bone mineral), and DMSA-chelatable lead (4.8-2,103 microg) levels among lead workers. Among lead workers, 9.9% (n = 79) were heterozygous for the ALAD(2) allele and there were no homozygotes. For VDR, 10.7% (n = 85) had the Bb genotype, and 0.5% (n = 4) had the BB genotype. Although the ALAD and VDR genes are located on different chromosomes, lead workers homozygous for the ALAD(1) allele were much less likely to have the VDR bb genotype (crude odds ratio = 0.29, 95% exact confidence interval = 0.06-0.91). In adjusted analyses, subjects with the ALAD(2) allele had higher blood lead levels (on average, 2.9 microg/dL, p = 0.07) but no difference in tibia lead levels compared with subjects without the allele. In adjusted analyses, lead workers with the VDR B allele had significantly (p < 0.05) higher blood lead levels (on average, 4.2 microg/dL), chelatable lead levels (on average, 37.3 microg), and tibia lead levels (on average, 6.4 microg/g) than did workers with the VDR bb genotype. The current data confirm past observations that the ALAD gene modifies the toxicokinetics of lead and also provides new evidence that the VDR gene does so as well. PMID- 11049815 TI - Acute effects of polychlorinated biphenyl-containing and -free transformer fluids on rat testicular steroidogenesis. AB - Polychlorinated biphenyl (PCB)-based transformer fluids belong to a class of environmentally persistent mixtures with known toxic effects. Here, we studied the acute effects of Askarel (which contains Aroclor 1260) and two substitute transformer fluids (the silicone oil-based DC561 and the mineral oil-based ENOL C) on rat testicular steroidogenesis. Single intraperitoneal (ip; 10 mg/kg body weight) or bilateral intratesticular (itt; 25 microg/testis) injections of Askarel markedly decreased serum androgen levels 24 hr after administration. In acute testicular cultures from these animals, chorionic gonadotropin-stimulated progesterone and androgen productions were severely attenuated. When itt was injected or added in vitro, Askarel inhibited 3ss-hydroxysteroid dehydrogenase (3ssHSD), stimulated 17[alpha]-hydroxylase/lyase (P450c17), and did not affect 17ss-hydroxysteroid dehydrogenase in testicular postmitochondrial fractions. The ip-injected Askarel did not affect 3ssHSD, but inhibited P450c17, suggesting that a more intensive metabolism of peripherally injected Askarel reduces the circulating levels of active ingredients below the threshold needed for inhibition of 3ssHSD and generates a derivative that inhibits P450c17. In contrast to Askarel, itt-injection (25 microg/testis) of DC561 and ENOL C did not affect in vivo and in vitro steroidogenesis. These findings show the acute effects of Askarel, but not silicone and mineral oils, on testicular steroidogenesis. PMID- 11049816 TI - The question of declining sperm density revisited: an analysis of 101 studies published 1934-1996. AB - In 1992 Carlsen et al. reported a significant global decline in sperm density between 1938 and 1990 [Evidence for Decreasing Quality of Semen during Last 50 Years. Br Med J 305:609-613 (1992)]. We subsequently published a reanalysis of the studies included by Carlsen et al. [Swan et al. Have Sperm Densities Declined? A Reanalysis of Global Trend Data. Environ Health Perspect 105:1228 1232 (1997)]. In that analysis we found significant declines in sperm density in the United States and Europe/Australia after controlling for abstinence time, age, percent of men with proven fertility, and specimen collection method. The declines in sperm density in the United States (approximately 1.5%/year) and Europe/Australia (approximately 3%/year) were somewhat greater than the average decline reported by Carlsen et al. (approximately 1%/year). However, we found no decline in sperm density in non-Western countries, for which data were very limited. In the current study, we used similar methods to analyze an expanded set of studies. We added 47 English language studies published in 1934-1996 to those we had analyzed previously. The average decline in sperm count was virtually unchanged from that reported previously by Carlsen et al. (slope = -0.94 vs. 0.93). The slopes in the three geographic groupings were also similar to those we reported earlier. In North America, the slope was somewhat less than the slope we had found for the United States (slope = -0.80; 95% confidence interval (CI), 1.37--0.24). Similarly, the decline in Europe (slope = -2.35; CI, -3.66--1.05) was somewhat less than reported previously. As before, studies from other countries showed no trend (slope = -0.21; CI, -2.30-1.88). These results are consistent with those of Carlsen et al. and our previous results, suggesting that the reported trends are not dependent on the particular studies included by Carlsen et al. and that the observed trends previously reported for 1938-1990 are also seen in data from 1934-1996. PMID- 11049817 TI - Effect of electromagnetic field exposure on chemically induced differentiation of friend erythroleukemia cells. AB - Whether exposure of humans to extremely low frequency electromagnetic fields (ELF EMF) can cause cancer is controversial and therefore needs further research. We used a Friend erythroleukemia cell line that can be chemically induced to differentiate to determine whether ELF-EMF could alter proliferation and differentiation in these cells in a manner similar to that of a chemical tumor promoter. Exposure of this cell line to 60 Hz ELF-EMF resulted in a dose dependent inhibition of differentiation, with maximal inhibition peaking at 40% and 40 mG (4 microT). ELF-EMF at 10 mG (1.0 microT) and 25 mG (2.5 microT) inhibited differentiation at 0 and 20%, respectively. ELF-EMF at 1.0 (100) and 10.0 G (1,000 microT) stimulated cell proliferation 50% above the sham-treated cells. The activity of telomerase, a marker of undifferentiated cells, decreased 100[times] when the cells were induced to differentiate under sham conditions, but when the cells were exposed to 0.5 G (50 microT) there was only a 10[times] decrease. In summary, ELF-EMF can partially block the differentiation of Friend erythroleukemia cells, and this results in a larger population of cells remaining in the undifferentiated, proliferative state, which is similar to the published results of Friend erythroleukemia cells treated with chemical-tumor promoters. PMID- 11049818 TI - Levels of seven urinary phthalate metabolites in a human reference population. AB - Using a novel and highly selective technique, we measured monoester metabolites of seven commonly used phthalates in urine samples from a reference population of 289 adult humans. This analytical approach allowed us to directly measure the individual phthalate metabolites responsible for the animal reproductive and developmental toxicity while avoiding contamination from the ubiquitous parent compounds. The monoesters with the highest urinary levels found were monoethyl phthalate (95th percentile, 3,750 ppb, 2,610 microg/g creatinine), monobutyl phthalate (95th percentile, 294 ppb, 162 microg/g creatinine), and monobenzyl phthalate (95th percentile, 137 ppb, 92 microg/g creatinine), reflecting exposure to diethyl phthalate, dibutyl phthalate, and benzyl butyl phthalate. Women of reproductive age (20-40 years) were found to have significantly higher levels of monobutyl phthalate, a reproductive and developmental toxicant in rodents, than other age/gender groups (p < 0.005). Current scientific and regulatory attention on phthalates has focused almost exclusively on health risks from exposure to only two phthalates, di-(2-ethylhexyl) phthalate and di-isononyl phthalate. Our findings strongly suggest that health-risk assessments for phthalate exposure in humans should include diethyl, dibutyl, and benzyl butyl phthalates. PMID- 11049820 TI - Prediction and assessment of the effects of mixtures of four xenoestrogens. AB - The assessment of mixture effects of estrogenic agents is regarded as an issue of high priority by many governmental agencies and expert decision-making bodies all over the world. However, the few mixture studies published so far have suffered from conceptual and experimental problems and are considered to be inconclusive. Here, we report the results of assessments of two-, three- and four-component mixtures of o,p'-DDT, genistein, 4-nonylphenol, and 4-n-octylphenol, all compounds with well-documented estrogenic activity. Extensive concentration response analyses with the single agents were carried out using a recombinant yeast screen (yeast estrogen screen, YES). Based on the activity of the single agents in the YES assay we calculated predictions of entire concentration response curves for mixtures of our chosen test agents assuming additive combination effects. For this purpose we employed the models of concentration addition and independent action, both well-established models for the calculation of mixture effects. Experimental concentration-response analyses revealed good agreement between predicted and observed mixture effects in all cases. Our results show that the combined effect of o,p'-DDT, genistein, 4-nonylphenol, and 4-n-octylphenol in the YES assay does not deviate from expected additivity. We consider both reference models as useful tools for the assessment of combination effects of multiple mixtures of xenoestrogens. PMID- 11049819 TI - Environmentally relevant xenoestrogen tissue concentrations correlated to biological responses in mice. AB - The effects of xenoestrogens have been extensively studied in rodents, generally under single, high-dose conditions. Using a continuous-release, low-dose system in ovariectomized mice, we correlated the estrogenic end points of uterine epithelial height (UEH) and vaginal epithelial thickness (VET) with concentrations of two organochlorine pesticide isomers in fat and blood. Silastic capsules containing a range of doses of either ss-hexachlorocyclohexane (ss-HCH) or o, p'-dichlorodiphenyltrichloroethane (o,p'-DDT) were implanted subcutaneously, and animals were killed after 1 week. Average blood levels achieved by the various doses were 4.2-620 ng/mL for o,p'-DDT and 5.0-300 ng/mL for ss-HCH. Fat concentrations of o,p'-DDT and ss-HCH correlated linearly to blood levels (o,p'-DDT, r(2) = 0.94; ss-HCH, r(2) = 0.83). Fat concentrations (nanograms per gram of tissue) were higher than blood concentrations (nanograms per milliliter) by 90 +/- 5- and 120 +/- 9-fold (mean +/- SE) for o, p'-DDT and ss-HCH, respectively. The VET ranged from 12 +/- 0.9 microm in controls to 114 +/ 8 microm in treated animals, and was correlated to blood levels of either treatment compound. The UEH ranged from an average of 7.7 +/- 0.3 microm in controls to 26 +/- 2 microm in high-dose o,p'-DDT-treated animals. The UEH was also correlated with ss-HCH concentration, but it plateaued at approximately 11 microm at the highest doses. The lowest blood concentrations that produced statistically significant increases in VET or UEH were 18 +/- 2 ng/mL o,p'-DDT and 42 +/- 4 ng/mL ss-HCH. These values are within the same order of magnitude of blood concentrations found in some human subjects from the general population, suggesting that human blood concentrations of these organochlorines may reach estrogenic levels. PMID- 11049821 TI - Environmental threats to the health of children: the Asian perspective. PMID- 11049822 TI - Early warning systems for hazardous biological agents in potable water. PMID- 11049823 TI - Evaluation and use of epidemiological evidence for environmental health risk assessment: WHO guideline document. AB - Environmental health risk assessment is increasingly being used in the development of environmental health policies, public health decision making, the establishment of environmental regulations, and research planning. The credibility of risk assessment depends, to a large extent, on the strength of the scientific evidence on which it is based. It is, therefore, imperative that the processes and methods used to evaluate the evidence and estimate health risks are clear, explicit, and based on valid epidemiological theory and practice. Epidemiological Evidence for Environmental Health Risk Assessment is a World Health Organization (WHO) guideline document. The primary target audiences of the guidelines are expert review groups that WHO (or other organizations) might convene in the future to evaluate epidemiological evidence on the health effects of environmental factors. These guidelines identify a set of processes and general approaches to assess available epidemiological information in a clear, consistent, and explicit manner. The guidelines should also help in the evaluation of epidemiological studies with respect to their ability to support risk assessment and, consequently, risk management. Conducting expert reviews according to such explicit guidelines would make health risk assessment and subsequent risk management and risk communication processes more readily understood and likely to be accepted by policymakers and the public. It would also make the conclusions reached by reviews more readily acceptable as a basis for future WHO guidelines and other recommendations, and would provide a more rational basis for setting priorities for future research. PMID- 11049826 TI - Drugged drinking water. PMID- 11049825 TI - The "new" genetics and mammalian cloning in environmental health research. PMID- 11049824 TI - Government laboratory worker with lung cancer: comparing risks from beryllium, asbestos, and tobacco smoke. AB - Occupational medicine physicians are frequently asked to establish cancer causation in patients with both workplace and non-workplace exposures. This is especially difficult in cases involving beryllium for which the data on human carcinogenicity are limited and controversial. In this report we present the case of a 73-year-old former technician at a government research facility who was recently diagnosed with lung cancer. The patient is a former smoker who has worked with both beryllium and asbestos. He was referred to the University of California, San Francisco, Occupational and Environmental Medicine Clinic at San Francisco General Hospital for an evaluation of whether past workplace exposures may have contributed to his current disease. The goal of this paper is to provide an example of the use of data-based risk estimates to determine causation in patients with multiple exposures. To do this, we review the current knowledge of lung cancer risks in former smokers and asbestos workers, and evaluate the controversies surrounding the epidemiologic data linking beryllium and cancer. Based on this information, we estimated that the patient's risk of lung cancer from asbestos was less than his risk from tobacco smoke, whereas his risk from beryllium was approximately equal to his risk from smoking. Based on these estimates, the patient's workplace was considered a probable contributing factor to his development of lung cancer. PMID- 11049827 TI - Controversy swirls around toilet-to-tap project. PMID- 11049828 TI - Minimizing mercury in medicine. PMID- 11049829 TI - Responsible care goes online. PMID- 11049830 TI - Reaching out to New York neighborhoods. PMID- 11049831 TI - Neural control of the kidney: functionally specific renal sympathetic nerve fibers. AB - The sympathetic nervous system provides differentiated regulation of the functions of various organs. This differentiated regulation occurs via mechanisms that operate at multiple sites within the classic reflex arc: peripherally at the level of afferent input stimuli to various reflex pathways, centrally at the level of interconnections between various central neuron pools, and peripherally at the level of efferent fibers targeted to various effectors within the organ. In the kidney, increased renal sympathetic nerve activity regulates the functions of the intrarenal effectors: the tubules, the blood vessels, and the juxtaglomerular granular cells. This enables a physiologically appropriate coordination between the circulatory, filtration, reabsorptive, excretory, and renin secretory contributions to overall renal function. Anatomically, each of these effectors has a dual pattern of innervation consisting of a specific and selective innervation by unmyelinated slowly conducting C-type renal sympathetic nerve fibers in addition to an innervation that is shared among all the effectors. This arrangement permits the maximum flexibility in the coordination of physiologically appropriate responses of the tubules, the blood vessels, and the juxtaglomerular granular cells to a variety of homeostatic requirements. PMID- 11049832 TI - Differential effects of inhaled nitric oxide and hyperoxia on pulmonary dysfunction in newborn guinea pigs. AB - This study tested the hypothesis that inhaled nitric oxide (NO) and combined NO and hyperoxia will result in less pulmonary dysfunction and delay onset of respiratory signs compared with hyperoxia-exposed newborn guinea pigs (GPs). GPs were exposed to room air (n = 14), 95% O(2) (n = 36), 20 parts per million (ppm) NO (n = 14), or combined 20 ppm NO and 95% O(2) (NO/O(2), n = 13) for up to 5 days. Data evaluated included latency interval for onset of respiratory distress, pressure volume curves, lung histology, and bronchoalveolar lavage (BAL) polymorphonuclear cells (PMNs), proteolytic activity, and total protein. NO exposed GPs did not develop respiratory distress and had no evidence of pulmonary dysfunction. O(2)-exposed GPs developed respiratory distress after 1-5 days (median 4.0) vs. 3-5 days (median 5.0) for NO/O(2) exposure (P < 0.05). BAL from O(2)-exposed GPs showed increased PMNs compared with NO/O(2)-exposed GPs. O(2)- and NO/O(2)-exposed GPs had comparable reduced lung volumes, lung histology, and increased BAL proteinase activity and total protein. In summary 1) O(2) exposure resulted in multiple measures of pulmonary dysfunction in newborn GPs, 2) 5-day exposure to NO produced no noticeable respiratory effects and pulmonary dysfunction, and 3) short-term exposure ( pH(out)) stimulated the uptake of L-proline. PMID- 11049844 TI - PDGF-beta receptor expression and ventilatory acclimatization to hypoxia in the rat. AB - Activation of platelet-derived growth factor-beta (PDGF-beta) receptors in the nucleus of the solitary tract (nTS) modulates the late phase of the acute hypoxic ventilatory response (HVR) in the rat. We hypothesized that temporal changes in PDGF-beta receptor expression could underlie the ventilatory acclimatization to hypoxia (VAH). Normoxic ventilation was examined in adult Sprague-Dawley rats chronically exposed to 10% O(2), and at 0, 1, 2, 7, and 14 days, Northern and Western blots of the dorsocaudal brain stem were performed for assessment of PDGF beta receptor expression. Although no significant changes in PDGF-beta receptor mRNA occurred over time, marked attenuation of PDGF-beta receptor protein became apparent after day 7 of hypoxic exposure. Such changes were significantly correlated with concomitant increases in normoxic ventilation, i.e., with VAH (r: -0.56, P < 0.005). In addition, long-term administration of PDGF-BB in the nTS via osmotic pumps loaded with either PDGF-BB (n = 8) or vehicle (Veh; n = 8) showed that although no significant changes in the magnitude of acute HVR occurred in Veh over time, the typical attenuation of HVR by PDGF-BB decreased over time. Furthermore, PDGF-BB microinjections did not attenuate HVR in acclimatized rats at 7 and 14 days of hypoxia (n = 10). We conclude that decreased expression of PDGF-beta receptors in the dorsocaudal brain stem correlates with the magnitude of VAH. We speculate that the decreased expression of PDGF-beta receptors is mediated via internalization and degradation of the receptor rather than by transcriptional regulation. PMID- 11049845 TI - Temperature-dependent development of cardiac activity in unrestrained larvae of the minnow Phoxinus phoxinus. AB - The minnow (Phoxinus phoxinus) was raised up to the stage of swim bladder inflation at temperatures between 10 degrees C and 25 degrees C, and the time of development significantly decreased at higher temperatures. Accordingly, initiation of cardiac activity was observed at day 2 in 25 degrees C animals and at day 4 in 12.5 degrees C animals. Only a minor increase in body mass was observed during the incubation period, and, at the end of the incubation period, animals raised at 25 degrees C did not have a significantly lower body mass compared with animals raised at 15 degrees C. Metabolic activity, determined as the rate of oxygen consumption of a larva, increased from 3.3 to 19.5 nmol/h during development at 15 degrees C and from 5.6 to 47.6 nmol/h during development at 25 degrees C. Heart rate showed a clear correlation to developmental stage as well as to developmental temperature, but at the onset of cardiac activity, diastolic ventricular volume and also stroke volume were higher at the lower temperatures. Furthermore, stroke volume increased with development, except for the group incubated at 12.5 degrees C, in which stroke volume decreased with development. Initial cardiac output showed no correlation to incubation temperature. Although metabolic activity increased severalfold during development from egg to the stage of swim bladder inflation at 15 degrees C and at 25 degrees C, weight-specific cardiac output increased only by approximately 40% with proceeding development. At 12.5 degrees C, cardiac output remained almost constant until opening of the swim bladder. The data support the notion that oxygen transport is not the major function of the circulatory system at this stage of development. The changes in heart rate with temperature appear to be due to the intrinsic properties of the pacemaker; there was no indication for a regulated response. PMID- 11049846 TI - Role of cyclooxygenase-2-derived metabolites and nitric oxide in regulating renal function. AB - The aim of this study was to examine the relative contribution of both cyclooxygenase (COX) isoforms in producing the prostaglandins (PG) involved in the regulation of renal function, when nitric oxide (NO) synthesis is reduced. In anesthetized dogs with reduction of NO synthesis, the renal effects of a nonisozyme-specific COX inhibitor (meclofenamate) were compared with those elicited by a selective COX-2 inhibitor (nimesulide) before and during an extracellular volume expansion (ECVE). Intrarenal N(G)- nitro-L-arginine methyl ester (L-NAME) infusion (1 microg x kg(-1) x min(-1); n = 6) did not elicit renal hemodynamic changes and reduced (P < 0.01) the renal excretory response to ECVE. Intravenous nimesulide (5 microg x kg(-1) x min(-1); n = 6) did not modify renal hemodynamic and reduced (P < 0. 05) sodium excretion before ECVE. Simultaneous L NAME and nimesulide infusion (n = 7) elicited an increment (37%) in renal vascular resistance (RVR; P < 0.05) before ECVE and no hemodynamic changes during ECVE. The reduced excretory response elicited by L-NAME and nimesulide was similar to that found during L-NAME infusion. Finally, simultaneous L-NAME and meclofenamate infusion (10 microg x kg(-1) x min(-1); n = 7) induced an increase in RVR (91%, P < 0.05), a decrease in glomerular filtration rate (35%, P < 0.05), and a reduction of the renal excretory response to ECVE that was greater (P < 0.05) than that elicited by L-NAME alone. The results obtained support the notion that PG involved in regulating renal hemodynamic and excretory function when NO synthesis is reduced are mainly dependent on COX-1 activity. PMID- 11049847 TI - Potassium channels in primary cultures of seawater fish gill cells. I. Stretch activated K(+) channels. AB - Previous studies using the patch-clamp technique demonstrated the presence of a small conductance Cl(-) channel in the apical membrane of respiratory gill cells in primary culture originating from sea bass Dicentrarchus labrax. We used the same technique here to characterize potassium channels in this model. A K(+) channel of 123 +/- 3 pS was identified in the cell-attached configuration with 140 mM KCl in the bath and in the pipette. The activity of the channel declined rapidly with time and could be restored by the application of a negative pressure to the pipette (suction) or by substitution of the bath solution with a hypotonic solution (cell swelling). In the excised patch inside-out configuration, ionic substitution demonstrated a high selectivity of this channel for K(+) over Na(+) and Ca(2+). The mechanosensitivity of this channel to membrane stretching via suction was also observed in this configuration. Pharmacological studies demonstrated that this channel was inhibited by barium (5 mM), quinidine (500 microM), and gadolinium (500 microM). Channel activity decreased when cytoplasmic pH was decreased from 7.7 to 6.8. The effect of membrane distension by suction and exposure to hypotonic solutions on K(+) channel activity is consistent with the hypothesis that stretch-activated K(+) channels could mediate an increase in K(+) conductance during cell swelling. PMID- 11049848 TI - Potassium channels in primary cultures of seawater fish gill cells. II. Channel activation by hypotonic shock. AB - Previous studies performed on apical membranes of seawater fish gills in primary culture have demonstrated the existence of stretch-activated K(+) channels with a conductance of 122 pS. The present report examines the involvement of K(+) channels in ion transport mechanisms and cell swelling. In the whole cell patch clamp configuration, K(+) currents were produced by exposing cells to a hypotonic solution or to 1 microM ionomycin. These K(+) currents were inhibited by the addition of quinidine and charybdotoxin to the bath solution. Isotopic efflux measurements were performed on cells grown on permeable supports using (86)Rb(+) as a tracer to indicate potassium movements. Apical and basolateral membrane (86)Rb effluxes were stimulated by the exposure of cells to a hypotonic medium. During the hypotonic shock, the stimulation of (86)Rb efflux on the apical side of the monolayer was inhibited by 500 microM quinidine or 100 microM gadolinium but was insensitive to scorpion venom [Leirus quinquestriatus hebraeus (LQH)]. An increased (86)Rb efflux across the basolateral membrane was also reduced by the addition of quinidine and LQH venom but was not modified by gadolinium. Moreover, basolateral and apical membrane (86)Rb effluxes were not modified by bumetanide or thapsigargin. There is convincing evidence for two different populations of K(+) channels activated by hypotonic shock. These populations can be separated according to their cellular localization (apical or basolateral membrane) and as a function of their kinetic behavior and pharmacology. PMID- 11049849 TI - Development of a compartmental model of human zinc metabolism: identifiability and multiple studies analyses. AB - A compartmental model of zinc metabolism has been developed from stable isotope tracer studies of five healthy adults. Multiple isotope tracers were administered orally and intravenously, and the resulting enrichment was measured in plasma, erythrocytes, urine, and feces for as long as 3 wk. Data from total zinc measurements and model-independent calculations of various steady-state parameters were also modeled with the kinetic data. A structure comprised of 14 compartments and as many as 25 unknown kinetic parameters was developed to adequately model the data from each of the individual studies. The structural identifiability of the model was established using the GLOBI2 identifiability analysis software. Numerical identifiability of parameter estimates was evaluated using statistical data provided by SAAM. A majority of the model parameters was estimated with sufficient statistical certainty to be considered well determined. After the fitting of the model and data from the individual studies using SAAM/CONSAM, results were submitted to SAAM extended multiple studies analysis for aggregation into a single set of population parameters and statistics. The model was judged to be valid based on criteria described elsewhere. PMID- 11049850 TI - Increased ganglionic responses to substance P in hypertensive rats due to upregulation of NK(1) receptors. AB - Intravenous injection of substance P (SP) increases renal nerve firing and heart rate in spontaneously hypertensive rats (SHRs) and Wistar-Kyoto rats (WKYs) by stimulating sympathetic ganglia. Blood pressure is increased in SHRs but lowered in WKYs. This study assesses the role of neurokinin-1 (NK(1)) receptors in mediating the ganglion actions of SP. Rats for functional studies were anesthetized and then treated with chlorisondamine. Renal nerve, blood pressure, and heart rate responses to intravenous injection of the NK(1) receptor agonist GR-73632 were similar but less than those to equimolar doses of SP in SHRs. GR 73632 only slightly increased renal nerve firing and heart rate and lowered blood pressure in WKYs. The NK(1) receptor antagonist GR-82334 (200 nmol/kg iv) blocked the ganglionic actions of GR-73632 and the pressor response to SP in SHRs. It reduced the renal nerve and heart rate responses by 52 and 35%. This suggests that the pressor response to SP is mediated by ganglionic NK(1) receptors and that NK(1) receptors also have a prominent role in mediating the renal nerve and heart rate responses to SP. Quantitative autoradiography showed that NK(1) receptors are more abundant in the superior cervical ganglia of SHRs. RT-PCR showed increased abundance of NK(1) receptor mRNA in SHRs as well. These observations suggest that the greater ganglionic stimulation caused by SP in SHRs is due to upregulation of NK(1) receptors. PMID- 11049851 TI - Effect of locally applied drugs on the pH of luminal fluid in the endolymphatic sac of guinea pig. AB - The aim of the present work was to assess the effect of various drugs applied locally on the pH of the luminal fluid (pH(lum)) in guinea pig endolymphatic sac. pH(lum) and transepithelial potential, when measured in vivo by means of double barrelled pH-sensitive microelectrodes, were 7.06 +/- 0.08 and +6.1 +/- 0.34 mV (mean +/- SE; n = 84), respectively, which is consistent with a net acid secretion in the luminal fluid of the endolymphatic sac. Bafilomycin and acetazolamide increased and decreased, respectively, pH(lum). Amiloride, ethylisopropylamiloride, ouabain, and Schering 28080 had no effect on pH(lum). Results obtained with inhibitors of anionic transport systems were inconclusive; e.g., DIDS reduced pH(lum), whereas neither SITS nor triflocin had any effect. We conclude that bafilomycin-sensitive H(+)-ATPase activity accounts for the transepithelial acid gradient measured in the endolymphatic sac and that intracellular and membrane-bound carbonic anhydrase probably participates in regulating endolymphatic sac pH(lum). The relationship between acid pH, endolymph volume, and Meniere's disease remains to be further investigated. PMID- 11049852 TI - Endothelin and ET(A) receptors in long-term arterial pressure homeostasis in conscious nonhuman primates. AB - This study was designed to quantify the long-term contribution of endogenous endothelin-1 (ET-1) and ET(A) receptors to the regulation of arterial pressure under normal conditions in nonhuman primates. Therefore, mean arterial pressure (MAP) and heart rate were measured 24 h/day with the use of telemetry techniques in conscious cynomolgus monkeys under control conditions, during administration of an ET(A) selective receptor antagonist (ABT-627; 5 mg/kg, 2 times a day by mouth, 4 days), and a 6-day posttreatment period. Systemic ET(A) blockade reduced MAP (24 h) from 89 +/- 3 to 82 +/- 2 and 79 +/- 2 mmHg on days 1 and 4, respectively. Subsequently, MAP remained suppressed for 3 days posttreatment. Heart rate increased from 111 +/- 5 to 122 +/- 4 and 128 +/- 6 beats/min on days 1 and 4 of ABT-627, respectively, and remained above control for 3 days posttreatment. Plasma ET-1 concentration increased from 1.0 +/- 0.3 to 1.9 +/- 0.4 pg/ml in response to ABT-627 (day 4) but decreased to control values 4 days posttreatment. These data demonstrate a physiologically important role for endogenous ET-1 and ET(A) receptors in the long-term regulation of arterial pressure and plasma ET-1 levels in the conscious nonhuman primate. PMID- 11049853 TI - Ca(2+) binding to cardiac troponin C: effects of temperature and pH on mammalian and salmonid isoforms. AB - A reduction in temperature lowers the Ca(2+) sensitivity of skinned cardiac myofilaments but this effect is attenuated when native cardiac troponin C (cTnC) is replaced with skeletal TnC. This suggests that conformational differences between the two isoforms mediate the influence of temperature on contractility. To investigate this phenomenon, the functional characteristics of bovine cTnC (BcTnC) and that from rainbow trout, Oncorhynchus mykiss, a cold water salmonid (ScTnC), have been compared. Rainbow trout maintain cardiac function at temperatures cardioplegic to mammals. To determine whether ScTnC is more sensitive to Ca(2+) than BcTnC, F27W mutants were used to measure changes in fluorescence with in vitro Ca(2+) titrations of site II, the activation site. When measured under identical conditions, ScTnC was more sensitive to Ca(2+) than BcTnC. At 21 degrees C, pH 7.0, as indicated by K(1/2) (-log[Ca] at half-maximal fluorescence, where [Ca] is calcium concentration), ScTnC was 2.29-fold more sensitive to Ca(2+) than BcTnC. When pH was kept constant (7.0) and temperature was lowered from 37.0 to 21.0 degrees C and then to 7.0 degrees C, the K(1/2) of BcTnC decreased by 0.13 and 0.32, respectively, whereas the K(1/2) of ScTnC decreased by 0.76 and 0.42, respectively. Increasing pH from 7.0 to 7.3 at 21.0 degrees C increased the K(1/2) of both BcTnC and ScTnC by 0.14, whereas the K(1/2) of both isoforms was increased by 1.35 when pH was raised from 7.0 to 7.6 at 7.0 degrees C. PMID- 11049854 TI - Effects of exercise on mitogen- and stress-activated kinase signal transduction in human skeletal muscle. AB - Exercise/contraction is a powerful stimulator of mitogen-activated protein (MAP) kinase cascades in skeletal muscle. Little is known regarding the physiological activation of enzymes downstream of MAP kinase. We investigated whether acute exercise results in activation of mitogen- and stress-activated kinases (MSK) 1 and 2, p90 ribosomal S6 kinase (p90rsk), and MAP kinase-activated protein kinase 2 (MAPKAPK2). Muscle biopsies were obtained from healthy volunteers before, during, and after 60 min one-leg cycle ergometry, from exercising and resting legs. MSK1 and MSK2 activities were increased 400-500% and 200-300%, respectively, in exercised muscle (P < 0.05 vs. rest). A dramatic increase in activity of p90rsk (MAPKAPK1) (>2,500%), and to a lesser extent MAPKAP2 (300%), was noted with exercise (P < 0.05 vs. rest). MSK1, MSK2, p90rsk, and MAPKAP2 activities were sustained throughout exercise. Exercise-induced activation of these enzymes was limited to working muscle, indicating that local rather than systemic factors activate these signaling cascades. Thus physical exercise leads to activation of multiple enzymes downstream of MAP kinase. PMID- 11049855 TI - Functional morphology of the avian medullary cone. AB - The organization of the renal medulla of the Gambel's quail, Callipepla gambelii, kidney was examined to determine the number of loops of Henle and collecting ducts and the surface area occupied by the different nephron segments as a function of distance down the medullary cones. Eleven medullary cones were dissected from the kidneys of four birds, and the tissue was processed and sectioned for light microscopy. In addition, individual nephrons were isolated on which total loop thin descending segment and thick prebend segment lengths were measured. The results show no correlation between the absolute number of loops of Henle and the length of the medullary cones. The number of thick and thin limbs of Henle and collecting ducts decrease exponentially with distance toward the apex of the cones and the rate of decrease is similar for cones of different lengths. Initially there is a rapid decrease in the number of thin limbs of Henle, indicating that most nephrons do not penetrate the cones a great distance. Thick descending limbs of Henle (prebend segment) ranged in length from 50 to 770 microm, and there was little correlation with the total length of the loop of Henle. However, the length of the thin limb of Henle correlated well with total loop length. The cell surface areas of the limbs of the loop of Henle and the collecting ducts decreased toward the apex of the cones. PMID- 11049856 TI - Interaction of SK(Ca) channels and L-type Ca(2+) channels in catecholamine secretion in the rat adrenal gland. AB - We elucidated the interaction of small-conductance Ca(2+)-activated K(+) (SK(Ca)) channels and L-type Ca(2+) channels in muscarinic receptor-mediated control of catecholamine secretion in the isolated perfused rat adrenal gland. The muscarinic agonist methacholine (10-300 microM) produced concentration-dependent increases in adrenal output of epinephrine and norepinephrine. The SK(Ca) channel blocker apamin (1 microM) enhanced the methacholine-induced catecholamine responses. The facilitatory effect of apamin on the methacholine-induced catecholamine responses was not observed during treatment with the L-type Ca(2+) channel blocker nifedipine (3 microM) or Ca(2+)-free solution. Nifedipine did not affect the methacholine-induced catecholamine responses, but it inhibited the responses during treatment with apamin. The L-type Ca(2+) channel activator Bay k 8644 (1 microM) enhanced the methacholine-induced catecholamine responses, whereas the enhancement of the methacholine-induced epinephrine and norepinephrine responses were prevented and attenuated by apamin, respectively. These results suggest that SK(Ca) channels are activated by muscarinic receptor stimulation, which inhibits the opening of L-type Ca(2+) channels and thereby attenuates adrenal catecholamine secretion. PMID- 11049857 TI - Sympathetic activity in early renal posttransplantation hypertension in rats. AB - The contribution of elevated sympathetic activity to the development of renal posttransplantation hypertension was investigated. F1 hybrids (F1H) from spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) were transplanted with either an SHR or an F1H kidney and bilaterally nephrectomized. Three weeks after transplantation, sympathetic activity was assessed by measuring adrenal tyrosine hydroxylase (TH) mRNA content and recording splanchnic nerve activity (SNA) in conscious animals. To investigate the dependence of arterial pressure on sympathetic activity, animals were treated with the alpha(2) adrenoceptor agonist guanabenz intracerebroventricularly. Mean arterial pressure (MAP) was 143 +/- 4 mmHg in recipients of an SHR kidney (n = 15) versus 110 +/- 3 mmHg in recipients of an F1H kidney (n = 10; P < 0.001). Adrenal TH mRNA content was 1.93 +/- 0.15 fmol/microg total RNA in recipients of an SHR kidney versus 1.96 +/- 0.17 fmol/microg total RNA in recipients of an F1H kidney (not significant). SNA did not differ significantly between recipients of an SHR kidney (n = 8) and recipients of an F1H kidney (n = 7) in terms of frequency and amplitude of synchronized nerve discharges. In response to cumulative intracerebroventricular administration of 10 and 20 microg guanabenz, SNA fell to 51 +/- 5% of control in recipients of an SHR kidney versus 44 +/- 6% of control in recipients of an F1H kidney (not significant) accompanied by a slight fall in MAP in either group. The results suggest that elevated sympathetic activity is not a major contributor to the development of renal posttransplantation hypertension. PMID- 11049858 TI - Effects of neonatal handling on sympathoadrenal activity and body composition in adult male rats. AB - Neonatal handling permanently alters the hypothalamic-pituitary-adrenal (HPA) response to stress. Because the sympathetic nervous system (SNS) and adrenal medulla also participate in stress responses, the impact of daily handling between birth and weaning on SNS and adrenal medullary function was examined in adult rats using techniques of [(3)H]norepinephrine ([(3)H]NE) turnover and urinary catecholamine excretion. Handled animals exhibited a 23% reduction in [(3)H]NE turnover in heart and a 53% decrease in spleen. [(3)H]NE turnover in brown adipose tissue, stomach, and kidney did not differ between handled and nonhandled animals. In contrast, urinary epinephrine (Epi) excretion was significantly greater in handled rats in response to a 3-day fast than in nonhandled animals. Although body weight, weight gain in response to dietary enrichment with sucrose or lard, or body fat content did not differ in handled and nonhandled animals, handled rats displayed heavier abdominal fat depots than nonhandled animals, implying a difference in body fat distribution. Neonatal handling thus leads to decreased sympathetic activity within specific subdivisions of the SNS and, by contrast, to increased adrenal medullary responsiveness. PMID- 11049859 TI - Role of presympathetic C1 neurons in the sympatholytic and hypotensive effects of clonidine in rats. AB - The rostral ventrolateral medulla (RVLM) may play an important role in the sympatholytic and hypotensive effects of clonidine. The present study examined which type of presympathetic RVLM neuron is inhibited by clonidine, and whether the adrenergic presympathetic RVLM neurons are essential for clonidine-induced sympathoinhibition. In chloralose-anesthetized and ventilated rats, clonidine (10 microg/kg iv) decreased arterial pressure (116 +/- 6 to 84 +/- 2 mmHg) and splanchnic nerve activity (93 +/- 3% from baseline). Extracellular recording and juxtacellular labeling of barosensitive bulbospinal RVLM neurons revealed that most cells were inhibited by clonidine (26/28) regardless of phenotype [tyrosine hydroxylase (TH)-immunoreactive cells: 48 +/- 7%; non-TH-immunoreactive cells: 42 +/- 5%], although the inhibition of most neurons was modest compared with the observed sympathoinhibition. Depletion of most bulbospinal catecholaminergic neurons, including 76 +/- 5% of the rostral C1 cells, by microinjection of saporin anti-dopamine beta-hydroxylase into the thoracic spinal cord (levels T2 and T4, 42 ng. 200 nl(-1). side(-1)) did not alter the sympatholytic or hypotensive effects of clonidine. These data show that although clonidine inhibits presympathetic C1 neurons, bulbospinal catecholaminergic neurons do not appear to be essential for the sympatholytic and hypotensive effects of systemically administered clonidine. Instead, the sympatholytic effect of clonidine is likely the result of a combination of effects on multiple cell types both within and outside the RVLM. PMID- 11049860 TI - Different adrenal sympathetic preganglionic neurons regulate epinephrine and norepinephrine secretion. AB - Brain stimulation or activation of certain reflexes can result in differential activation of the two populations of adrenal medullary chromaffin cells: those secreting either epinephrine or norepinephrine, suggesting that they are controlled by different central sympathetic networks. In urethan-chloralose anesthetized rats, we found that antidromically identified adrenal sympathetic preganglionic neurons (SPNs) were excited by stimulation of the rostral ventrolateral medulla (RVLM) with either a short (mean: 29 ms) or a long (mean: 129 ms) latency. The latter group of adrenal SPNs were remarkably insensitive to baroreceptor reflex activation but strongly activated by the glucopenic agent 2 deoxyglucose (2-DG), indicating their role in regulation of adrenal epinephrine release. In contrast, adrenal SPNs activated by RVLM stimulation at a short latency were completely inhibited by increases in arterial pressure or stimulation of the aortic depressor nerve, were unaffected by 2-DG administration, and are presumed to govern the discharge of adrenal norepinephrine-secreting chromaffin cells. These findings of a functionally distinct preganglionic innervation of epinephrine- and norepinephrine-releasing adrenal chromaffin cells provide a foundation for identifying the different sympathetic networks underlying the differential regulation of epinephrine and norepinephrine secretion from the adrenal medulla in response to physiological challenges and experimental stimuli. PMID- 11049861 TI - Methysergide delays the decompensatory responses to severe hemorrhage by activating 5-HT(1A) receptors. AB - Central administration of the serotonin receptor ligand methysergide delays the decompensatory response to hypotensive hemorrhage. This study was performed to determine the receptor subtype that mediates this effect. Lateral ventricular (LV) injection of methysergide (40 microg) delayed the hypotensive, bradycardic, and sympathoinhibitory responses to blood withdrawal (1.26 ml/min) in conscious rats. The response was quantified, in part, as the blood volume withdrawal that produced a 40-mmHg fall in blood pressure. The delayed hypotensive response produced by methysergide (8.2 +/- 0.2 vs. 5.6 +/- 0.2 ml, P < 0.01) was reversed by the 5-hydroxytryptamine (HT)(1A) antagonist WAY-100635 (30 microg iv: 6.7 +/- 0.4 ml, P < 0. 01; 100 microg iv: 5.6 +/- 0.1 ml, P < 0.01). LV injection of the 5-HT(1A) agonist (+)-8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT) also delayed the hypotensive (10 microg: 8.6 +/- 0.3, P < 0.01; 20 microg: 9.2 +/- 0.3 ml, P < 0.01), bradycardic, and sympathoinhibitory responses to hemorrhage. WAY 100635 (10 microg iv) completely reversed the effects of 8-OH-DPAT (20 microg: 5.4 +/- 0.3 ml). Neither selective blockade of 5-HT(2) receptors nor stimulation of 5-HT(1B/1D) receptors had any effect on hemorrhage responses. These data indicate that methysergide stimulates 5-HT(1A) receptors to delay the decompensatory responses to hemorrhage. PMID- 11049862 TI - Activity and responsiveness of the renin-angiotensin system in the aging rat. AB - The systemic renin-angiotensin system (RAS) is suppressed in normal aging, but the activity of the tissue RAS is not well defined. We examined the systemic and intrarenal RAS status of aging normal rats and responses to suppression and stimulation of the production of endogenous ANG II. Studies were performed in young (3 mo) and early aging (15 mo) male Sprague-Dawley rats. Angiotensin converting enzyme inhibitors modestly decreased mean arterial pressure (MAP) in young (3 mo) and early aging (15 mo) rats and limited proteinuria in the older rats. There were no significant age-related effects on renal function or on endogenous RAS activity. Intravenous infusion of the precursor ANG I led to comparable increases in MAP in younger and older rats. In contrast, the renal effects (reduction in glomerular filtration and plasma flow rates) were exaggerated in the older animals. Intrarenal arterial ANG I did not affect MAP in any group. In young rats, there were no significant hemodynamic effects in either the ipsilateral (infused) or the contralateral (noninfused) kidney. In the older rats, both kidneys had a significant fall in renal renal plasma flow rate (RPF) with left renal arterial infusion of ANG I. Accordingly, these studies early in the course of aging found only subtle changes in the activity, responsiveness, and metabolism of the RAS. Thus early aging is associated with a modest but important increase in sensitivity to RAS stimulation. PMID- 11049863 TI - Ingested water equilibrates isotopically with the body water pool of a shorebird with unrivaled water fluxes. AB - We investigated the applicability of (2)H to measure the amount of body water (TBW) and water fluxes in relation to diet type and level of food intake in a mollusk-eating shorebird, the Red Knot (Calidris canutus). Six birds were exposed to eight experimental indoor conditions. Average fractional (2)H turnover rates ranged between 0. 182 day(-1) (SD = 0.0219) for fasting birds and 7.759 day(-1) (SD = 0.4535) for birds feeding on cockles (Cerastoderma edule). Average TBW estimates obtained with the plateau method were within the narrow range of 75.9 85.4 g (or between 64.6 and 70.1% of the body mass). Those obtained with the extrapolation method showed strong day-to-day variations (range 55.7-83.7 g, or between 49.7 and 65.5%). Average difference between the two calculation methods ranged between 0.6% and 36.3%, and this difference was strongly negatively correlated with water flux rate. Average water influx rates ranged between 15.5 g/day (fasting) and 624.5 g/day (feeding on cockles). The latter value is at 26.6 times the allometrically predicted value and is the highest reported to date. Differences in (2)H concentrations between the blood and feces (i.e., biological fractionation) were small but significant (-3.4% when fed a pellet diet, and 1.1% for all the other diets), and did not relate to the rate of water flux (chi(2)(1) = 0.058, P < 0.81). We conclude that the ingested water equilibrated rapidly with the body water pool even in an avian species that shows record water flux rates when living on ingested marine bivalves. PMID- 11049864 TI - Capsaicin-treated rats permanently overingest low- but not high-concentration sucrose solutions. AB - The effect of capsaicin-induced chemical ablation of visceral afferents on 1-h liquid sucrose consumption was investigated in food-deprived rats. We first show that although 10% sucrose is permanently overconsumed by capsaicin-treated (CAPs) compared with vehicle-treated (VEHs) control rats, 40% sucrose is only overconsumed during the first but not subsequent 1-h exposures. Furthermore, one group of CAPs lost the overconsumption response at 20% when exposed to progressively increasing sucrose concentrations of 10-40%, and another group recovered the overconsumption response at 10% when exposed to a series of decreasing concentrations. Control rats ingested relatively constant volumes of sucrose over the range of 10, 15, and 20%, resulting in significantly different energy intakes. In contrast, CAPs generally showed a concentration-dependent decrease in volume intake, resulting in relatively constant energy intake. These results suggest that capsaicin-sensitive visceral afferents, likely from gastric distension and other preabsorptive sensors, provide major control over volume ingested. In the absence of these signals, rats initially overconsume, but rapidly learn to use other signals from capsaicin-resistant preabsorptive or postabsorptive sites, to control future intake. This redundant satiety system appears to be sensitive to the osmotic value or caloric content of the unfamiliar food, but only if this is above a threshold of about 15% sucrose. PMID- 11049865 TI - Chronic hypoxia increases the NO contribution of acetylcholine vasodilation of the fetal guinea pig heart. AB - To investigate the effect of chronic hypoxia (HPX) on vasodilation of the fetal heart, we exposed pregnant guinea pigs to room air or 12% O(2) for 4, 7, or 10 days. We excised hearts from anesthetized fetuses (60 +/- 3 days; 65-day gestation = term) and measured changes in both the coronary artery pressure of the isolated constant-flow preparation and endothelial nitric oxide synthase (eNOS) mRNA of fetal ventricles. Dilator responses to cumulative addition (10(-9) 10(-5) M) of acetylcholine and sodium nitroprusside in prostaglandin F(2alpha) (5 x 10(-6) M)-constricted hearts were similar among normoxia (NMX), 4-, 7-, and 10 day HPX (control). Nitro-L-arginine (L-NA, 10(-4)M), a NOS inhibitor, inhibited maximal acetylcholine dilation of hearts exposed to 10-day HPX greater than NMX, 4-, and 7-day HPX. Hypoxia (after 7 and 10 days) increased eNOS mRNA of fetal ventricles compared with NMX and 4-day HPX. 4-Aminopyridine (3 mM), a voltage dependent K(+)-channel inhibitor, inhibited acetylcholine- but not sodium nitroprusside-induced dilation of NMX and 10-day HPX hearts to a similar magnitude. Glibenclamide (10(-5) M), an ATP-sensitive K(+)-channel inhibitor, had no effect on vasodilation. We conclude that chronic HPX increases the contribution of NO but does not alter K(+)-channel activation in response to acetylcholine-stimulated coronary dilation. Thus increases in NO production via upregulation of eNOS gene expression may be an adaptive response to chronic HPX in the fetal coronary circulation. PMID- 11049866 TI - Central angiotensin receptor blockade impairs thermolytic and dipsogenic responses to heat exposure in rats. AB - The effect of central angiotensin AT(1) receptor blockade on thermoregulation and water intake after heat exposure was investigated. Rats were placed in a chamber heated to 39 +/- 1 degrees C for 60 min and then returned to their normal cage (at 22 degrees C), and water intake was measured for 120 min. Artificial cerebrospinal fluid (5 microl) was injected intracerebroventricularly 60 min before heat exposure in five control rats. Colonic temperature increased from 37.22 +/- 0.21 to 40.68 +/- 0.31 degrees C after 60 min. In six rats injected intracerebroventricularly with 10 microg of the AT(1) antagonist losartan, colonic temperature increased from 37.41 +/- 0.27 to 41.72 +/- 0.28 degrees C after 60 min. This increase was significantly greater than controls (P < 0.03). Losartan-treated rats drank 1.1 +/- 0.4 ml of water compared with 5.9 +/- 0.77 ml (P < 0.002) drank by control animals, despite a similar body weight loss in the two groups. Central losartan did not inhibit the drinking response to intracerebroventricular carbachol in heated rats, suggesting that losartan treatment did not nonspecifically depress behavior. We conclude that central angiotensinergic mechanisms have a role in both thermoregulatory cooling in response to heat exposure and also the ensuing water intake. PMID- 11049867 TI - Renal nerve inhibition by central NaCl and ANG II is abolished by lesions of the lamina terminalis. AB - The lamina terminalis is situated in the anterior wall of the third ventricle and plays a major role in fluid and electrolyte homeostasis and cardiovascular regulation. The present study examined whether the effects of intracerebroventricular infusion of hypertonic saline and ANG II on renal sympathetic nerve activity (RSNA) were mediated by the lamina terminalis. In control, conscious sheep (n = 5), intracerebroventricular infusions of 0.6 M NaCl (1 ml/h for 20 min) and ANG II (10 nmol/h for 30 min) increased mean arterial pressure (MAP) by 6 +/- 1 (P < 0.001) and 14 +/- 3 mmHg (P < 0.001) and inhibited RSNA by 80 +/- 6 (P < 0.001) and 89 +/- 7% (P < 0.001), respectively. Both treatments reduced plasma renin concentration (PRC). Intracerebroventricular infusion of artificial cerebrospinal fluid (1 ml/h for 30 min) had no effect. In conscious sheep with lesions of the lamina terminalis (n = 6), all of the responses to intracerebroventricular hypertonic saline and ANG II were abolished. In conclusion, the effects of intracerebroventricular hypertonic saline and ANG II on RSNA, PRC, and MAP depend on the integrity of the lamina terminalis, indicating that this site plays an essential role in coordinating the homeostatic responses to changes in brain Na(+) concentration. PMID- 11049868 TI - Inhibition of poly(ADP-ribose) polymerase attenuates ischemic renal injury in rats. AB - The enzyme, poly(ADP-ribose) polymerase (PARP), effects repair of DNA after ischemia-reperfusion (I/R) injury to cells in nerve and muscle tissue. However, its activation in severely damaged cells can lead to ATP depletion and death. We show that PARP expression is enhanced in damaged renal proximal tubules beginning at 6-12 h after I/R injury. Intraperitoneal administration of PARP inhibitors, benzamide or 3-amino benzamide, after I/R injury accelerates the recovery of normal renal function, as assessed by monitoring the levels of plasma creatinine and blood urea nitrogen during 6 days postischemia. PARP inhibition leads to increased cell proliferation at 1 day postinjury as assessed by proliferating cell nuclear antigen and improves the histopathological appearance of kidneys examined at 7 days postinjury. Furthermore, inhibition of PARP increases levels of ATP measured at 24 h postischemia compared with those in vehicle-treated animals. Our data indicate that PARP activation is a part of the cascade of molecular events that occurs after I/R injury in the kidney. Although caution is advised, transient inhibition of PARP postischemia may constitute a novel therapy for acute renal failure. PMID- 11049869 TI - Testosterone receptor blockade after trauma and hemorrhage attenuates depressed adrenal function. AB - Although the testosterone receptor antagonist flutamide restores the depressed immune function in males after trauma and hemorrhage, it remains unknown whether this agent has any salutary effects on adrenal function. To study this, male rats underwent laparotomy and were bled to and maintained at a blood pressure of 40 mmHg until 40% of the shed blood volume was returned in the form of Ringer lactate. Animals were then resuscitated and flutamide (25 mg/kg body wt) was administered subcutaneously. Plasma adrenocorticotropic hormone (ACTH) and corticosterone, as well as adrenal corticosterone and cAMP were measured 20 h after resuscitation. In additional animals, ACTH was administered and ACTH induced corticosterone release and adrenal cAMP were determined. The results indicate that adrenal contents of corticosterone and cAMP were significantly decreased and morphology was altered after hemorrhage. Administration of flutamide improved corticosterone content, restored cAMP content, and attenuated adrenal morphological alterations. Flutamide also improved the diminished ACTH induced corticosterone release and adrenal cAMP response at 20 h after hemorrhage and resuscitation. Furthermore, the diminished corticosterone response to ACTH stimulation in the isolated adrenal preparation was improved with flutamide. These results suggest that flutamide is a useful adjunct for improving adrenal function in males following trauma and hemorrhage. PMID- 11049870 TI - Hemodynamic responses to static and dynamic muscle contractions at equivalent workloads. AB - We tested the hypothesis that static contraction causes greater reflex cardiovascular responses than dynamic contraction at equivalent workloads [i.e., same tension-time index (TTI), holding either contraction time or peak tension constant] in chloralose-anesthetized cats. When time was held constant and tension was allowed to vary, dynamic contraction of the hindlimb muscles evoked greater increases (means +/- SE) in mean arterial pressure (MAP; 50 +/- 7 vs. 30 +/- 5 mmHg), popliteal blood velocity (15 +/- 3 vs. 5 +/- 1 cm/s), popliteal venous PCO(2) (15 +/- 3 vs. 3 +/- 1 mmHg), and a greater decrease in popliteal venous pH (0.07 +/- 0.01 vs. 0.03 +/- 0.01), suggesting greater metabolic stimulation during dynamic contraction. Similarly, when peak tension was held constant and time was allowed to vary, dynamic contraction evoked a greater increase in blood velocity (13 +/- 1 vs. -1 +/- 1 cm/s) without causing any differences in other variables. To investigate the reflex contribution of mechanoreceptors, we stretched the hindlimb dynamically and statically at the same TTI. A larger reflex increase in MAP during dynamic stretch (32 +/- 8 vs. 24 +/- 6 mmHg) was observed when time was held constant, indicating greater mechanoreceptor stimulation. However, when peak tension was held constant, there were no differences in the reflex cardiovascular response to static and dynamic stretch. In conclusion, at comparable TTI, when peak tension is variable, dynamic muscle contraction causes larger cardiovascular responses than static contraction because of greater chemical and mechanical stimulation. However, when peak tensions are equivalent, static and dynamic contraction or stretch produce similar cardiovascular responses. PMID- 11049871 TI - Na(+)/Ca(2+)-exchange activity regulates contraction and SR Ca(2+) content in rainbow trout atrial myocytes. AB - We have used the whole cell configuration of the patch-clamp technique to measure sarcolemmal Ca(2+) transport by the Na(+)/Ca(2+) exchanger (NCX) and its contribution to the activation and relaxation of contraction in trout atrial myocytes. In contrast to mammals, cell shortening continued, increasing at membrane potentials above 0 mV in trout atrial myocytes. Furthermore, 5 microM nifedipine abolished L-type Ca(2+) current (I(Ca)) but only reduced cell shortening and the Ca(2+) carried by the tail current to 66 +/- 5 and 67 +/- 6% of the control value. Lowering of the pipette Na(+) concentration from 16 to 10 or 0 mM reduced Ca(2+) extrusion from the cell from 2.5 +/- 0.2 to 1.0 +/- 0.2 and 0.5 +/- 0.06 amol/pF. With 20 microM exchanger inhibitory peptide (XIP) in the patch pipette Ca(2+) extrusion 20 min after patch break was 39 +/- 8% of its initial value. With 16, 10, and 0 mM Na(+) in the pipette, the sarcoplasmic reticulum (SR) Ca(2+) content was 47 +/- 4, 29 +/- 6, and 10 +/- 3 amol/pF, respectively. Removal of Na(+) from or inclusion of 20 microM XIP in the pipette gradually eliminated the SR Ca(2+) content. Whereas I(Ca) was the same at -10 or +10 mV, Ca(2+) extrusion from the cell and the SR Ca(2+) content at -10 mV were 65 +/- 7 and 80 +/- 4% of that at +10 mV. The relative amount of Ca(2+) extruded by the NCX (about 55%) and taken up by the SR (about 45%) was, however, similar with depolarizations to -10 and +10 mV. We conclude that modulation of the NCX activity critically determines Ca(2+) entry and cell shortening in trout atrial myocytes. This is due to both an alteration of the transsarcolemmal Ca(2+) transport and a modulation of the SR Ca(2+) content. PMID- 11049872 TI - Aging and baroreflex control of RSNA and heart rate in rats. AB - Aging is associated with altered autonomic control of cardiovascular function, but baroreflex function in animal models of aging remains controversial. In this study, pressor and depressor agent-induced reflex bradycardia and tachycardia were attenuated in conscious old (24 mo) rats [57 and 59% of responses in young (10 wk) Wistar rats, respectively]. The intrinsic heart rate (HR, 339 +/- 5 vs. 410 +/- 10 beats/min) was reduced in aged animals, but no intergroup differences in resting mean arterial blood pressure (MAP, 112 +/- 3 vs. 113 +/- 5 mmHg) or HR (344 +/- 9 vs. 347 +/- 9 beats/min) existed between old and young rats, respectively. The aged group also exhibited a depressed (49%) parasympathetic contribution to the resting HR value (vagal effect) but preserved sympathetic function after intravenous methylatropine and propranolol. An implantable electrode revealed tonic renal sympathetic nerve activity (RSNA) was similar between groups. However, old rats showed impaired baroreflex control of HR and RSNA after intravenous nitroprusside (-0.63 +/- 0. 18 vs. -1.84 +/- 0.4 bars x cycle(-1) x mmHg(-1) x s(-1)). Therefore, aging in rats is associated with 1) preserved baseline MAP, HR, and RSNA, 2) impaired baroreflex control of HR and RSNA, and 3) altered autonomic control of resting HR. PMID- 11049873 TI - Human skeletal muscle exercise metabolism following an expedition to mount denali. AB - Chronic exposure to high altitude is known to result in changes in the mechanisms regulating O(2) delivery to the contracting muscle. However, the effects of acclimatization on metabolism in the contracting muscle cell remain unclear. In this study, we have investigated the hypothesis that acclimatization would result in a closer coupling between ATP utilization and ATP production and that the improved energy state would be accompanied by a reorganization of the metabolic pathways consisting of an increased oxidative and decreased glycolytic potential. Five men, mean age of 28 +/- 2 (SE) yr, performed a standardized, two-stage submaximal cycling task in normoxia for 20 min at each of 59 and 74% peak O(2) consumption before and 3-4 days after returning from a 21-day expedition to Mount Denali (6,194 m). Acclimatization was without effect in altering the resting values of the adenine nucleotides (ATP, ADP, AMP), inosine monophosphate (IMP), or phosphocreatine (PCr) in the vastus lateralis. During exercise (40 min) after acclimatization compared with preacclimatization, PCr was not as depressed (33.2 +/- 7.1 vs. 40.6 +/- 5.4 mmol/kg dry wt) and IMP (0.289 +/- 0.11 vs. 0. 131 +/- 0.03 mmol/kg dry wt) and lactate (26.1 +/- 6.2 vs. 18.6 +/- 8.8 mmol/kg dry wt) in contracting muscle were not as elevated (P < 0.05). Although no effect of acclimatization was observed for the maximal activity (mol. kg protein(-1). h( 1)) of citrate synthase (4. 76 +/- 0.44 vs. 4.94 +/- 0.45), lactate dehydrogenase was increased by 13% (36.5 +/- 2.6 vs. 41.2 +/- 3.1, P < 0.05). It is concluded that acclimatization results in an improved energy state in the contracting muscle when tested under normoxic conditions; however, these effects are not associated with a higher oxidative potential or a lower glycolytic potential as hypothesized. PMID- 11049874 TI - Short-term modulation of the exercise ventilatory response in goats: effects of 8 OH-DPAT and MPPI. AB - Increased respiratory dead space increases the exercise ventilatory response, a response known as short-term modulation (STM). We hypothesized that STM results from a spinal, serotonin (5-HT)-dependent mechanism. Because 5-HT(1A) autoreceptors on caudal brain stem raphe neurons inhibit 5-HT release, we hypothesized that 5-HT(1A)-receptor agonists would inhibit, whereas 5-HT(1A) receptor antagonists would enhance, STM. Ventilatory and arterial blood-gas measurements were made at rest and during exercise (4.0-4.5 km/h, 5% grade) in goats with the respiratory mask alone or with increased dead space (0.20-0.25 liter), before and after intravenous administration of the 5-HT(1A)-receptor agonist 8-hydroxy-2-(di-n-propylamino)tetralin (8-OH-DPAT; 0.1 mg/kg) or the antagonist 4-iodo-N-(2-[4-(methoxyphenyl)-1-piperazinyl]ethyl)-N-2-pyridinylbenz amide (MPPI; 0.08 mg/kg). 8-OH-DPAT increased the slope of the arterial PCO(2) vs. metabolic CO(2) production relationship and decreased the ventilation vs. metabolic CO(2) production relationship during exercise with increased dead space (not with the mask alone), indicating an impairment of STM. In contrast, MPPI had minimal effects on any measured variable. Although nonspecific effects of 8-OH DPAT cannot be ruled out, impaired STM is consistent with the hypothesis that STM requires active raphe serotonergic neurons and 5-HT release. PMID- 11049875 TI - Characterization of RyR1-slow, a ryanodine receptor specific to slow-twitch skeletal muscle. AB - Two distinct skeletal muscle ryanodine receptors (RyR1s) are expressed in a fiber type-specific manner in fish skeletal muscle (11). In this study, we compare [(3)H]ryanodine binding and single channel activity of RyR1-slow from fish slow twitch skeletal muscle with RyR1-fast and RyR3 isolated from fast-twitch skeletal muscle. Scatchard plots indicate that RyR1-slow has a lower affinity for [(3)H]ryanodine when compared with RyR1-fast. In single channel recordings, RyR1 slow and RyR1-fast had similar slope conductances. However, the maximum open probability (P(o)) of RyR1-slow was threefold less than the maximum P(o) of RyR1 fast. Single channel studies also revealed the presence of two populations of RyRs in tuna fast-twitch muscle (RyR1-fast and RyR3). RyR3 had the highest P(o) of all the RyR channels and displayed less inhibition at millimolar Ca(2+). The addition of 5 mM Mg-ATP or 2.5 mM beta, gamma-methyleneadenosine 5'-triphosphate (AMP-PCP) to the channels increased the P(o) and [(3)H]ryanodine binding of both RyR1s but also caused a shift in the Ca(2+) dependency curve of RyR1-slow such that Ca(2+)-dependent inactivation was attenuated. [(3)H]ryanodine binding data also showed that Mg(2+)-dependent inhibition of RyR1-slow was reduced in the presence of AMP-PCP. These results indicate differences in the physiological properties of RyRs in fish slow- and fast-twitch skeletal muscle, which may contribute to differences in the way intracellular Ca(2+) is regulated in these muscle types. PMID- 11049876 TI - Effects of prenatal dexamethasone on spatial learning and response to stress is influenced by maternal factors. AB - The present study investigated the effect of prenatal dexamethasone (Dex) exposure on early perinatal events, hippocampal function, and response to stress. Pregnant rats received Dex in the evening water (2.5 microg/ml) or tap water (Veh) from gestational day 15 until delivery. On the day of parturition, pups were randomized, cross-fostered, and reduced to eight or nine per dam. Four groups resulted: Veh-Veh (offspring exposed to Veh in utero, rearing mother treated with Veh during gestation), Veh-Dex, Dex-Veh, and Dex-Dex. Spatial visual memory was evaluated with the Morris water maze. The corticosterone response to restraint stress was examined, and the expression of hippocampal glucocorticoid and mineralocorticoid receptors mRNA was determined by in situ hybridization. Exposure to Dex caused restlessness in mothers, low birth weights, and poor weight gain in the offspring. The Dex-Dex males had impaired spatial learning, inability to rapidly terminate the adrenocortical response to stress, and decreased hippocampal glucocorticoid receptor (GR) mRNA expression. In contrast, Dex-exposed animals reared by Veh-treated mothers had adequate spatial learning, enhanced glucocorticoid feedback, and increased hippocampal GR mRNA. We conclude that the environment provided by a healthy mother during the postnatal period can prevent the detrimental effects of prenatal Dex administration on cognition, GR mRNA expression of the hippocampus, and the quality of the stress response. PMID- 11049877 TI - Carotid and aortic baroreflexes of the rat: I. Open-loop steady-state properties and blood pressure variability. AB - To characterize the baroreflex in central nervous system-intact neuromuscular blocked rats, we measured the vascular and cardiac responses and compared direct stimulation of the aortic depressor nerve (ADN) with a capacitance electrode (differentially activating either A or A + C fibers) to carotid sinus pressure with a micro-balloon (SINUS). One-thousand-two-hundred-ninety-seven open-loop measurements of systolic blood pressure (SBP), heart rate, venous pressure (VBP), and mesenteric (msBF), femoral (fmBF), and skin (skBF) blood flow were completed; the linear range of the effects was determined for each response and stimulus mode. The rats were sinoaortic denervated (SAD). The open-loop stimulation effect was very stable; e.g., the mean effect of 790 ADN stimulations during >7 days was -9.8 mmHg, with an average drift of +0.001 mmHg/h. In contrast, there was large variability of the SBP baseline (e.g., SD = +/-10.9), which was due to SAD (+/ 6.3 to +/-16.3 mmHg, t = -13. 9, df = 4, P < 0.0002) and was reversed by ganglionic block (+/-10.8 to +/- 2.9 mmHg, t = -12.9, df = 3, P < 0.001). The ADN stimuli produced larger depressor responses than sinus stimuli (-66 vs. -45 mmHg); all component responses paralleled the magnitude of the SBP effect, except interbeat interval (IBI), for which the ADN DeltaIBI was approximately 10 times that of SINUS. For all stimuli, fmBF increased and msBF did not. Mesenteric and femoral vascular conductance both increased, whereas VBP decreased and skBF followed SBP. We found that for all baroreflex response components, with the exception of SINUS-elicited DeltaIBI, there was an orderly, substantially linear, relationship between stimulus strength and response magnitude. PMID- 11049878 TI - Carotid and aortic baroreflexes of the rat: II. Open-loop frequency response and the blood pressure spectrum. AB - To determine the relationship between blood pressure (BP) variability and the open-loop frequency domain transfer function (TF) of the baroreflexes, we measured the pre- and postsinoaortic denervation (SAD) spectra and the effects of periodic and step inputs to the aortic depressor nerve and isolated carotid sinus of central nervous system-intact, neuromuscular-blocked (NMB) rats. Similar to previous results in freely moving rats, SAD greatly increased very low frequency (VLF) (0.01-0.2 Hz) systolic blood pressure (SBP) noise power. Step response frequency measurements for SBP; interbeat interval (IBI); venous pressure; mesenteric, femoral, and skin blood flow; and direct modulation analyses of SBP showed that only VLF variability could be substantially attenuated by an intact baroreflex. The -3-dB frequency for SBP is 0.035-0.056 Hz; femoral vascular conductance is similar to SBP, but mesenteric vascular conductance has a reliably lower and IBI has a reliably higher -3-dB point. The overall open-loop transportation lag, of which or = 133 micromol/L were also separately analyzed. Evaluated patients (n = 1502, +/-EACA; 581/905, 16 exclusions) included 233 with CrPre > or = 133 micromol/L (+/-EACA; 98/135). Multivariate analyses confirmed several known risk factors, but no association between DCrCl and EACA in all patients (P: = 0.66), and the subgroup with CrPre > or = 133 micromol/L (P: = 0.42). IMPLICATIONS: In a large population of primary Coronary Artery Bypass Graft including a subset with preoperative renal dysfunction, there were no postoperative reductions in creatinine clearance attributable to epsilon-aminocaproic (EACA) administration. This retrospective study suggests that moderate epsilon-aminocaproic acid dosing during cardiac surgery is safe for the kidney; however, this inference is based on a single marker of renal dysfunction and requires prospective confirmation using a variety of tests of renal function. PMID- 11049889 TI - Thromboelastography as a perioperative measure of anticoagulation resulting from low molecular weight heparin: a comparison with anti-Xa concentrations. AB - Low molecular weight heparin (LMWH) is commonly used to prevent postoperative thromboembolism. Currently, there is no convenient test to measure the degree of anticoagulation from LMWH. This prospective study examines the relationship of thromboelastography and serum anti-Xa concentration in patients treated with enoxaparin. Twenty-four adult patients scheduled for orthopedic surgery using epidural anesthesia were enrolled. Epidural catheters were removed the morning after surgery before the commencement of subcutaneous enoxaparin 30 mg twice daily. Venous blood samples were obtained at 1) the induction of anesthesia (baseline), 2) immediately before the third dose of enoxaparin postoperatively (Day 2-trough), 3) 4 h after the third dose postoperatively (Day 2-peak), and 4) immediately before the fifth dose postoperatively (Day 3-trough). Whole blood samples were obtained for thromboelastography, activated clotting time, and anti Xa level analyses at each of the four time intervals. At the four sample intervals, the r time (mean +/- SEM). (20 +/- 1, 25 +/- 2, 51 +/- 6, 31 +/- 3 mm) and the k time (9 +/- 0. 7, 12 +/- 1, 27 +/- 5, 14 +/- 2 mm) of the thromboelastograph were significantly correlated with the expected peak and trough levels of LMWH and serum anti-Xa levels (P: < 0.05). At the Day 3-trough, thromboelastograph r times exceeded the normal range in 6 of 25 patients (25%). Prolongation of r time and k time on postoperative Day 3 may indicate an exaggerated response to LMWH. Thromboelastography is a test that could potentially correlate with the degree of anticoagulation produced by low molecular weight heparin. IMPLICATIONS: Thromboelastography is a test that could potentially correlate with the degree of anticoagulation produced by low molecular weight heparin. The r time from the thromboelastogram correlates with serum anti-Xa concentration. PMID- 11049890 TI - Insulin reverses bupivacaine-induced cardiac depression in dogs. AB - We tested the hypothesis that an insulin infusion would effectively treat bupivacaine-induced cardiac depression in dogs. In 24 mongrel dogs anesthetized with pentobarbital (5 mgkg(-1)h(-1), IV), 0.5% bupivacaine was administrated at a rate of 0.5 mgkg(-1)min(-1) until the mixed venous oxygen saturation decreased to 60% or less. The bupivacaine infusion induced a decrease in mean arterial pressure, cardiac output, and heart rate. The dogs were randomly assigned to one of four groups after the end of bupivacaine infusion. The Control (C, n = 6) and Glucose (G, n = 6) groups received an IV infusion of normal saline (2 mL/kg) and glucose (2 mL/kg of 50% dextrose in water) for 15 min, respectively. The Insulin Glucose (IG, n = 6) group received an IV bolus of regular insulin (1 U/kg), plus a glucose infusion (2 mL/kg of 50% dextrose in water) for 15 min. The Insulin Glucose-Potassium (IGK, n = 6) group received the same dose of insulin and glucose as the IG group, plus potassium (1-3 mEqkg(-1)h(-1)). Mean arterial pressure, cardiac output, heart rate, and mixed venous oxygen saturation recovered toward baseline level more rapidly in the IG and IGK groups than in the C group (within 5 min versus more than 20 min). These results suggest that the infusion of insulin and glucose might reverse bupivacaine-induced cardiac depression in dogs. IMPLICATIONS: We found that insulin and glucose rapidly reversed hemodynamic abnormality in dogs with bupivacaine-induced cardiac depression. This study implies a possible clinical application of insulin treatment for bupivacaine-induced cardiac depression. PMID- 11049891 TI - Ventricular arrhythmias with or without programmed electrical stimulation after incremental overdosage with lidocaine, bupivacaine, levobupivacaine, and ropivacaine. AB - It is unclear whether the mechanism of death from local anesthetic (LA) intoxication is primarily a consequence of cardiac arrhythmias or myocardial contractile depression, and whether LAs might differ in this susceptibility to these two mechanisms. By using programmable electrical stimulation (PES) protocols in anesthetized, ventilated dogs, we compared the arrhythmogenic potential of bupivacaine (BUP), ropivacaine (ROP), levobupivacaine (LBUP), and lidocaine (LIDO). Open-chest dogs were randomized to receive escalating incremental infusions of the four local anesthetics until cardiovascular collapse. We assumed a concentration relationship of 4:1 for LIDO/BUP, LBUP, and ROP. The effective refractory period did not change significantly until the dose increment corresponding to target concentrations of 8 and 32 microg/mL for BUP, LBUP, ROP, and LIDO, respectively. Thirty percent to 50% increases in effective refractory period occurred in surviving dogs at this dose. The incidence of spontaneous or PES-induced ventricular tachycardia and ventricular fibrillation did not differ among groups. Compared with LIDO, the incidence of PES-induced extrasystoles was more frequent for BUP- and LBUP-treated dogs (P: < 0.05). ROP treated dogs did not differ from LIDO-treated dogs with respect to PES-induced extrasystoles. At the dose increment preceding cardiovascular collapse, all LAs produced significant increases in heart rate and reductions in blood pressure compared with their respective baseline values. The incidence of programmable electrical stimulation-induced ventricular tachycardia and fibrillation with BUP does not differ from the incidence that occurs with the single S:(-) enantiomers LBUP and ROP, providing further evidence against stereoselective arrhythmogenesis as a primary component of local anesthetic-induced cardiotoxicity. IMPLICATIONS: Progressive bupivacaine intoxication in anesthetized, ventilated dogs does not produce early arrhythmogenic events. The incidence of programmable electrical stimulation-induced ventricular tachycardia and fibrillation with bupivacaine does not differ from the incidence that occurs with the single S:(-) enantiomers levobupivacaine and ropivacaine, providing further evidence against stereoselective arrhythmogenesis as a primary component of local anesthetic induced cardiotoxicity. PMID- 11049892 TI - Dexmedetomidine and hemodynamic responses to simulated hemorrhage in experimental heart failure. AB - alpha(2)-Adrenoreceptor agonists may counteract the increased basal sympathetic nervous activity in patients with congestive heart failure (CHF), but they may also compromise reflex responses to hypovolemia. We have tested responses to simulated hemorrhage (central hypovolemia) after IV dexmedetomidine in normal animals and in experimental chronic CHF. Rabbits (n = 14) were treated with IV doxorubicin (or control saline) for 8 weeks inducing biventricular dilatation and myocardial damage. Cardiac output (CO) was measured continuously with a transit time Doppler implanted on the ascending aorta. Progressive inflation of a cuff around the inferior vena cava (simulated hemorrhage) was used to reduce cardiac index at a constant rate. Arterial baroreceptor-mediated vasoconstrictor and heart rate responses were tested with repeated cuff inflations. Although resting CO was reduced in CHF, the blood pressure and heart rate changes with dexmedetomidine were not exaggerated. The slope of the vasoconstrictor response to graded hypovolemia was attenuated by dexmedetomidine with an earlier onset of decompensation. There was no added effect of CHF on the response until the dose of dexmedetomidine was sufficient to reduce resting CO in addition to arterial blood pressure and heart rate. IMPLICATIONS: As an adjunct to anesthesia, dexmedetomidine may be useful in reducing basal sympathetic nervous activity. This study in experimental animals suggests this may be achieved without compromising protective responses to decreased blood volume. PMID- 11049893 TI - Perianesthetic risks and outcomes of pheochromocytoma and paraganglioma resection. AB - Pheochromocytomas and paragangliomas are often surgically curable. However, resection of these tumors can be life threatening. We undertook this study to determine the frequency of, and risk factors for, perioperative complications in patients undergoing resection of pheochromocytoma or paraganglioma. We retrospectively reviewed the medical records of patients during 1983-1996 who underwent surgical resection of catecholamine-secreting pheochromocytoma or paraganglioma. Preoperative risk factors, adverse intraoperative events, and complications occurring in the 30 days after operation were recorded. Blood pressures were collected from manual records. The ranked sum test and Fisher's exact test were used for analyses. Adverse perioperative events or complications occurred in 45 of 143 patients (31.5%; exact 95% confidence interval, 24.0% to 39.8%). Of these 45 patients, 41 experienced one or more adverse intraoperative events. The most common adverse event was sustained hypertension (36 patients). There were no perioperative deaths, myocardial infarctions, or cerebrovascular events. Preoperative factors univariately associated with adverse perioperative events included larger tumor size (P: = 0.007), prolonged duration of anesthesia (P: = 0.015), and increased levels of preoperative urinary catecholamines and catecholamine metabolites: vanillylmandelic acid (P: = 0.019), metanephrines (P: = 0.004), norepinephrine (P: = 0. 014), and epinephrine (P: = 0.004). Despite premedication of most patients with phenoxybenzamine and a beta-adrenergic blocker, varying degrees of intraoperative hemodynamic lability occurred. IMPLICATIONS: Few patients who had pheochromocytoma or paraganglioma resection experienced significant perioperative morbidity and none died in the largest retrospective study on this topic to date. This study confirms the very good perioperative outcomes demonstrated in smaller studies on this high-risk population, and identifies several risk factors for adverse outcomes. PMID- 11049894 TI - Tranexamic acid reduces blood loss in total hip replacement surgery. AB - Intraoperatively administered, tranexamic acid (TA) does not reduce bleeding in total hip replacement (THR). Therefore, its prophylactic use was attempted in the present study because this has been shown to be more effective in cardiac surgery. We investigated 40 patients undergoing THR in a prospective, randomized, double-blinded study. Twenty patients received TA given in two bolus doses of 10 mg/kg each, the first just before surgical incision and the second 3 h later. In addition, a continuous infusion of TA, 1.0 mg. kg(-1). h(-1) for 10 h, was given after the first bolus dose. The remaining 20 patients formed a control group. Both groups used preoperative autologous blood donation and intraoperative autotransfusion. Intraoperative bleeding was significantly less (P: = 0.001) in the TA group compared with the control group (630 +/- 220 mL vs 850 +/- 260 mL). Postoperative drainage bleeding was correspondingly less (P: = 0.001) (520 +/- 280 vs 920 +/- 410 mL). Up to 10 h postoperatively, plasma D-dimer concentration was halved in the TA group compared with the control group. One patient in each group had an ultrasound-verified late deep vein thrombosis. In conclusion, we found TA, administrated before surgical incision, to be efficient in reducing bleeding during THR. IMPLICATIONS: In a prospective, double-blinded study of 40 patients undergoing total hip replacement, the preoperative administration of tranexamic acid reduced bleeding by 35%, probably by decreasing induced fibrinolysis. Whether tranexamic acid therapy can replace predonation of autologous blood or intraoperative autotransfusion requires further study. PMID- 11049895 TI - Severe hypotension in a patient receiving pemoline during general anesthesia. AB - IMPLICATIONS: This case reports hypotension under general anesthesia in a patient taking pemoline. Vigilance for unexpected hypotension is important in patients who are treated with psychostimulants. If hypotension occurs, vasopressors that act directly on adrenergic receptors should be used. PMID- 11049896 TI - An unusual presentation of atrial septal defect in a patient undergoing total hip arthroplasty. AB - IMPLICATIONS: Atrial septal defect is usually diagnosed and surgically repaired in childhood. We present a case of previously asymptomatic atrial septal defect that presented during total hip arthroplasty during general anesthesia in a 72-yr old woman. Intraoperative transesophageal echocardiography assisted in the diagnosis and in determining the appropriate treatment. PMID- 11049897 TI - Reversible tricuspid valve obstruction during removal of renal cell carcinoma with intracardiac tumor extension. AB - IMPLICATIONS: Transesophageal echocardiography was used to identify and guide management of reversible tricuspid valve obstruction by a tumor mass during surgical removal of a renal cell carcinoma. PMID- 11049898 TI - Epidural catheter and increased prothrombin time after right lobe hepatectomy for living donor transplantation. AB - IMPLICATIONS: Donor right hepatic lobectomy for the purpose of living liver transplantation may be associated with postoperative abnormalities in tests of clotting function. This study explores the possible causes and anesthetic implications of this phenomenon. PMID- 11049899 TI - The incidence of bradycardia during endoscopic third ventriculostomy. AB - The incidence of bradycardia during endoscopic third ventriculostomy (ETV) is unknown. In an attempt to determine that incidence, we studied 49 pediatric patients with obstructive hydrocephalus who underwent ETV during general anesthesia. The median age was 54.5 mo (range 1-108 mo) and the median weight was 12.2 kg (range 2.4-22 kg). The heart rate was measured continuously in which four stages were identified for data analysis. Stage A is the preoperative phase, stage B is 5 min before perforating the floor of the third ventricle, stage C during perforation, and stage D after perforating the floor of the third ventricle. Three readings were recorded at each stage, then averaged. The mean values of the heart rate at stages A, B, C, and D were 146 +/- 27, 151 +/- 26, 87 +/- 32, and 143 +/- 24 bpm respectively. A significant decrease in the heart rate was determined in stage C compared with stage B (P: < 0.05). The incidence of bradycardia was 41%. Alerting the surgeon to perforate the floor of the third ventricle or withdraw the scope away from it was sufficient to resolve the bradycardia. We concluded that serious bradycardia might occur during ETV, mostly because of mechanical factors and can be resolved without medications. IMPLICATIONS: The use of endoscopy for treating pediatric patients with increased intracranial pressure is a new surgical procedure. These patients require general anesthesia with continuous heart rate monitoring. We have observed a high incidence of decrease in heart rate. If a decrease in heart rate occurs, alerting the surgeon to speed the procedure would be an effective treatment. PMID- 11049900 TI - Pressure control ventilation: three anesthesia ventilators compared using an infant lung model. AB - We compared three ventilators-Servo 900C (Siemens Medical Systems, Danvers, MA), Aestiva 3000 (Datex-Ohmeda, Madison, WI), and NAD 6000 (North American Drager, Telford, PA)-set to deliver pressure control ventilation using an infant test lung model. Ventilator settings were selected to test "near-maximum" settings that would be used for a neonatal patient (peak inspiratory pressure [PIP] 30 cm H(2)O) or older child (PIP 60 cm H(2)O). When adjusted for set inspiratory pressure and compliance, the average tidal volume (V(t)) produced by the NAD 6000 was 5.8 mL less than the Servo 900C (P: = 0. 103), and the average V(t) produced by the Aestiva 3000 was 18.9 mL less than the Servo 900C (P: < 0.001). The Servo 900C generated increased peak pressures, tending to overshoot the set maximum inflating pressures, especially during rapid respiratory rates with decreased inspiratory times. The Aestiva 3000 did not achieve the set PIP during testing conditions of decreased inspiratory times, and the NAD 6000 was not greatly affected by changes in inspiratory time. All three ventilators measured expiratory V(t) to be larger than the actual V(t) delivered to the lung; however, the NAD 6000 was more accurate. IMPLICATIONS: There are differences in performance of ventilators when set to deliver pressure control ventilation to an infant test lung model. PMID- 11049901 TI - Halothane and sevoflurane decrease norepinephrine-stimulated glucose transport in neonatal cardiomyocyte. AB - Catecholamine regulates myocardial glucose use. However, the effect of inhaled anesthetics on myocardial glucose transport stimulated by catecholamine is unclear. We studied the effect of halothane and sevoflurane on uptake of 2 deoxyglucose stimulated by norepinephrine in neonatal cardiomyocytes and the mechanism that modulates glucose transport. We studied the effects of halothane and sevoflurane on norepinephrine (NE)-stimulated glucose uptake and the effects of halothane and sevoflurane on glucose uptake stimulated by W7 (a calcium releasing agent), phorbol 12 myristate-13-acetate (a protein kinase C agonist), and LiCl. Sevoflurane decreased NE-stimulated glucose uptake from 63.7 +/- 7.0 to 41.2 +/- 3.7 pmol h(-1) mg protein(-1), and halothane also attenuated NE stimulated glucose uptake to 37.8 +/- 5.7 pmol h(-1) mg protein(-1). W7 at 10 micromol/L increased glucose uptake from 16.4 +/- 1.4 to 41.2 +/- 3. 4 pmol h(-1) mg protein(-1). The stimulation was inhibited in the presence of 0.8 mmol/L sevoflurane and 0.58 mmol/L halothane to 23.9 +/- 3.7 and 25.6 +/- 3.6 pmol h(-1) mg protein(-1), respectively. Halothane and sevoflurane did not significantly affect the glucose uptake stimulated by 1 nmol/L insulin, 10 micromol/L PMA, or 10 mmol/L LiCl. We conclude that halothane and sevoflurane decrease NE-stimulated glucose uptake through decrease in intracellular calcium in cardiomyocytes. IMPLICATIONS: The effect of inhaled anesthetics on myocardial glucose uptake during administration of catecholamine is unclear. The myocardial glucose uptake is stimulated not only by catecholamine, but also by insulin, protein kinase C, and increase of intracellular calcium. We examined the effects of halothane and sevoflurane on glucose uptake. PMID- 11049902 TI - Hypermagnesemia in a pediatric patient. AB - IMPLICATIONS: A high blood magnesium level is a rare but life-threatening event. This case report describes an accidental overdose of magnesium that occurred in an 18-month-old girl during a routine operation. The effects of high magnesium levels and its treatment are explained in detail. PMID- 11049903 TI - Part I: propofol, thiopental, sevoflurane, and isoflurane-A randomized, controlled trial of effectiveness. AB - When compared with thiopental and isoflurane, propofol and sevoflurane are associated with a faster return to wakefulness after anesthesia. Yet their wider usage in inpatient surgery has been restrained by concerns regarding their acquisition costs and by lack of studies demonstrating improved patient outcome. We randomly allocated 453 adult surgical inpatients to one of four anesthetic regimens (thiopental-isoflurane, propofol-isoflurane, propofol induction and maintenance, or sevoflurane induction and maintenance) and measured their rate and quality of recovery. We found no significant differences in the rate and quality of recovery between groups. Propofol was associated with more pain on injection (P: < 0. 0005), but less cough during induction (P: = 0.003), and less early postoperative nausea and vomiting (P: = 0.003). We could not detect any significant advantages with propofol and sevoflurane, when compared with thiopental and isoflurane in adults undergoing elective inpatient surgery. IMPLICATIONS: Propofol and sevoflurane do not offer any significant advantages over thiopental and isoflurane in adults undergoing elective inpatient surgery. PMID- 11049904 TI - Part II: total episode costs in a randomized, controlled trial of the effectiveness of four anesthetics. AB - Newer anesthetics promise improved clinical outcomes, but usually come at a higher price per dose. Previous studies have found few economic benefits in the immediate postoperative period, but have hypothesized that earlier recovery may lead to lower costs for the whole episode of hospitalization. This study uses cost data for patients enrolled in a randomized, controlled clinical trial comparing four anesthetics to test whether the higher costs of the newer anesthetics would be offset against decreased use of other hospital resources. Five hundred general surgery patients were randomly assigned to one of four anesthetic regimens. Estimates from the hospital's patient costing system were used, with validated cost records for a subset of 360 patients. Five patients admitted to the intensive care unit or requiring prolonged hospitalization skewed the distribution of costs, but none of these complications could be attributed to anesthesia. No significant differences were found on length of stay, mean episode cost, operating room costs, ward costs, or readmission rate within 3 mo. The study was not powered to sufficiently show differences in intensive care unit admission or other uncommon outcomes. Patient quality of recovery did not vary among groups, but neither patient willingness-to-pay nor satisfaction were directly measured. IMPLICATIONS: Propofol and sevoflurane do not offer any significant economic advantages over thiopental and isoflurane in adults undergoing elective inpatient surgery. PMID- 11049905 TI - Longer-term diabetic patients have a more frequent incidence of nosocomial infections after elective gastrectomy. AB - Diabetes mellitus (DM) is one of the risk factors for the development of postoperative nosocomial infections in surgical patients. We conducted this retrospective study to elucidate the perioperative risk factors for postoperative nosocomial infections in diabetic patients undergoing elective gastrectomy. Chart review was performed on diabetic and nondiabetic patients undergoing elective gastrectomy for gastric malignancy from January 1992 through April 1999. Fourteen of the 83 diabetic patients, and 23 of the 284 nondiabetic patients developed postoperative nosocomial infections. Statistical comparisons of multiple variables were made between patients with and without postoperative nosocomial infections. In diabetic patients, univariate analysis showed that longer-term DM (especially longer than 10 yr) was associated with a significantly increased risk for postoperative nosocomial infections. Multiple logistic regression analysis showed that DM lasting longer than 10 yr was an independent risk factor for postoperative nosocomial infections (odds ratio, 6.8; 95% confidence interval, 1.7 to 27.1). In nondiabetic patients, similar analysis showed that age was an independent risk factor for postoperative nosocomial infections. We conclude that patients with longer-term DM had a significantly greater incidence of postoperative nosocomial infections after elective gastrectomy. IMPLICATIONS: Postoperative nosocomial infection is one of the major problems in diabetic patients. This study demonstrated that postoperative nosocomial infections were more common in patients undergoing elective gastrectomy if they had diabetes mellitus longer than 10 yr. PMID- 11049906 TI - Early onset of regional intestinal ischemia can be detected with carbon dioxide tension measurement inside the peritoneal cavity. AB - Methods for detecting regional gastrointestinal ischemia are rare. An early detection of ischemia in the stomach or ileum can be achieved by the continuous intramucosal PCO(2) (PiCO(2)) measurement in the region. However, physiological consideration suggests that the placement of a fiberoptic CO(2) sensor in the peritoneal cavity should yield comparable results. We tested the hypothesis that a continuous PCO(2) measurement in the peritoneal cavity allows the early detection of regional intestinal ischemia. A laparotomy was performed in six pigs (54.7 +/- 3.7 kg) with a tourniquet being placed around respective vessels to allow complete ischemia of a 2. 75-m part of the ileum. A fiberoptic CO(2) sensor (PiCO(2)-ileum) was placed intraluminally in the ileum outside this segment. A second fiberoptic CO(2) sensor to measure intraperitoneal PCO(2) (i. p.-PCO(2)) was placed inside the peritoneal cavity in close vicinity to the ischemic gut segment. Gastric PiCO(2) was determined by using air tonometry. After baseline measurements, ileal ischemia was induced for 180 min followed by a 30-min reperfusion period. Statistics were performed with a Friedman test followed by Wilcoxon Analysis with P: < 0.01 considered significant. With the onset of local ileal ischemia, a sudden increase in i.p.-PCO(2) from 48.9 (45. 0-51.5) mm Hg (mean and 25-75 percentiles) to 94.3 (87.9-95.5; P: < 0.01) mm Hg was observed. Gastric PiCO(2) (49.0 [47.5-51.0]/53.5 [49. 0-54.0] mm Hg), and ileal PiCO(2) (56.4 [44.6-57.0]/54.3 [46.1-57.8] mm Hg) did not change. With reperfusion, the i.p.-PCO(2) decreased but stayed above baseline values. IMPLICATIONS: Unless systemic changes are induced, regional intestinal perfusion deficits cannot be detected with a PCO(2) measurement in the gastric lumen. In pigs, an occlusion of blood flow to an isolated gut segment resulted in a significant increase in intraperitoneal CO(2) tension. Thus, the measurement of intraperitoneal PCO(2) could allow the early detection of regional intestinal ischemia. PMID- 11049907 TI - Dynamic cardiocirculatory control during propofol anesthesia in mechanically ventilated patients. AB - We evaluated dynamic cardiovascular control by spectral analytical methods in 20 young adults anesthetized with propofol (2.5 mg/kg, followed by continuous infusion of 0.1 mg/kg/min) and in an awake control group during cyclic stimulation of the carotid baroreceptors via sinusoidal neck suction at 0.2 Hz (baroreflex response mediated mainly by vagal activity) and at 0.1 Hz (baroreflex response mediated by vagal and sympathetic activity). During anesthesia and mechanical ventilation at 0.25 Hz, major underdampened hemodynamic oscillations occurred at 0.055 +/- 0.012 Hz. The response of RR intervals to baroreceptor stimulation at 0.2 Hz was markedly decreased during anesthesia (median of transfer function magnitude between neck suction and RR intervals 3% of the awake control). Blood pressure response to baroreceptor stimulation at 0.1 Hz was significantly decreased during anesthesia to 26% (systolic blood pressure), and 44% (diastolic blood pressure) of the awake control. There was a significant delay in baroreflex effector responses during anesthesia. Our results demonstrate a markedly depressed vagally mediated heart rate response and an impaired blood pressure response to cyclic baroreceptor stimulation during propofol anesthesia in mechanically ventilated patients. The disturbed baroreflex control is accompanied by an irregular dynamic behavior of cardiovascular regulation, indicating a decreased stability of the control system. IMPLICATIONS: An irregular dynamic behavior of the cardiovascular control system, associated with an impaired baroreflex control of heart rate and blood pressure, can be observed during propofol anesthesia in mechanically ventilated subjects. PMID- 11049908 TI - The efficacy of hemodynamic and T wave criteria for detecting intravascular injection of epinephrine test doses in anesthetized adults: a dose-response study. AB - Recent studies have shown that an epidural test dose containing 15 microg of epinephrine has a sensitivity and specificity of 100% for detecting intravascular injection based on the systolic blood pressure (SBP) (positive if > or =15-mm Hg increase) and the T wave criteria (positive if > or =0.1 mV and 25% decrease in amplitude), whereas the modified heart rate (HR) criterion (positive if > or =10 bpm increase) produced uncertain results in sevoflurane-anesthetized adults. Because a fractional dose of the test dose may be injected intravascularly in actual clinical situations, we designed this study to determine, in a dose related manner, the efficacy and minimum effective dose of epinephrine based on those hemodynamic and the T wave criteria. Eighty healthy adult patients were randomly assigned to one of four groups according to a simulated IV test dose under 2% end-tidal sevoflurane and nitrous oxide anesthesia after endotracheal intubation (n = 20 each). The saline group received 3 mL of normal saline IV; the epinephrine-15 group received 3 mL of 1.5% lidocaine containing 15 microg of epinephrine (1); and the epinephrine-10 and -5 groups received 2 and 1 mL of the test dose of the identical components, respectively. HR, SBP, and lead II of the electrocardiograph were recorded continuously for 5 min after the IV injection of the study drug. Sensitivities and specificities of 100% were obtained based on the HR and the SBP criteria only if 15 microg of epinephrine was injected IV, whereas sensitivities and specificities of 100% were obtained based on both T wave criteria after 15 and 10 microg of epinephrine was injected IV. Two blinded observers were able to detect all T wave changes in patients who received 15, 10, and 5 microg of epinephrine IV, resulting in 100% efficacy (P: < 0.05 versus HR and SBP criteria). We conclude that minimum effective epinephrine doses for detecting accidental intravascular injection are 15 microg on the HR and the SBP criteria, and 10 microg on both T wave criteria, and that observing T wave changes may detect even smaller (5 microg) doses of epinephrine injected IV in adult patients anesthetized with sevoflurane and nitrous oxide. IMPLICATIONS: To determine whether an epidural catheter is in a blood vessel, an epidural test dose containing 15 microg of epinephrine is used. We found that a decrease in T wave amplitude appears to be more sensitive than heart rate and systolic blood pressure change for detecting accidental intravascular injection of a small dose of epinephrine-containing test dose in sevoflurane-anesthetized patients. PMID- 11049909 TI - The use of intravenous atropine after a saline infusion in the prevention of spinal anesthesia-induced hypotension in elderly patients. AB - We investigated the efficacy of IV atropine for preventing spinal anesthesia induced hypotension in elderly patients. Seventy-five patients undergoing transurethral prostate or bladder surgery were randomized to receive either placebo (n = 25), atropine 5 microg/kg (small-dose atropine, n = 25) or atropine 10 microg/kg (large-dose atropine, n = 25) after the induction of spinal anesthesia. All the patients received an IV infusion of 10 mL/kg 0.9% normal saline over 10 min before the induction of anesthesia. The systolic blood pressure decreased in all three groups after spinal anesthesia. There was a significant increase in the mean heart rate in both atropine groups as compared to the placebo group (placebo group: 78 bpm, 95% confidence interval [CI]: 76.6 78.5; small-dose atropine group: 86 bpm, 95% CI 83.9-88.8; large-dose atropine group: 97 bpm, 95% CI 94.5-100.3; P: = 0.001). There was a significant decrease in the incidence of hypotension in patients who received atropine (placebo group: 76%, small-dose atropine group: 52%, large-dose atropine group: 40%, P: = 0.03). The mean dose of ephedrine required was significantly decreased in the atropine groups (placebo group: 12.2 mg [SD= 10.5], small-dose atropine group: 7.4 mg [SD= 10.0], large-dose atropine group: 5.4 mg [SD= 8.7 mg], P: = 0.048). The total amount of IV fluid and number of patients requiring metaraminol in addition to 30 mg of ephedrine were not significantly different among the three groups. Significant side effects, such as confusion, ST segment changes or angina were not detected in any of the patients. We conclude that IV atropine may be a useful supplement to the existing methods in preventing hypotension induced by spinal anesthesia. IMPLICATIONS: IV atropine increases heart rate in a dose-dependent manner in elderly patients undergoing spinal anesthesia. It reduces the incidence of hypotension and the dose of ephedrine required. Small-dose atropine may be a useful supplement in preventing spinal anesthesia-induced hypotension in elderly patients. PMID- 11049910 TI - The effect of erythromycin, fluvoxamine, and their combination on the pharmacokinetics of ropivacaine. AB - We studied the effect of fluvoxamine and erythromycin on the pharmacokinetics of ropivacaine in a double-blinded, randomized, four-way cross-over study. Eight healthy volunteers ingested daily 1500 mg erythromycin for 6 days, 100 mg fluvoxamine for 5 days (Days 2-6), both erythromycin and fluvoxamine, or placebo. On Day 6, each subject received a single dose of 0.6 mg/kg ropivacaine IV over 30 min. Ropivacaine, 3-hydroxyropivacaine, and 2',6'-pipecoloxylidide in venous plasma and urine samples were measured for up to 12 h and 24 h, respectively. Fluvoxamine increased the area under the drug plasma concentration-time curve (AUC) of ropivacaine 3.7-fold (P: < 0.001), prolonged the elimination half-life (t(1/2)) from 2.3 to 7.4 h (P: < 0.01), and decreased the clearance by 77% (P: < 0.001). Erythromycin alone had only a minor effect on the pharmacokinetics of ropivacaine. However, when compared with fluvoxamine alone, the combination of fluvoxamine and erythromycin further increased the area under the drug plasma concentration-time curve and t(1/2) of ropivacaine by 50% (P: < 0.01). We conclude that inhibition of CYP1A2 by fluvoxamine considerably reduces elimination of ropivacaine. Concomitant use of fluvoxamine and CYP3A4 inhibitor erythromycin further increases plasma ropivacaine concentration by decreasing its clearance. IMPLICATIONS: Clinicians should be aware of the possibility of increased toxicity of ropivacaine when used together with inhibitors of CYP1A2. Concomitant use of CYP1A2 and CYP3A4 inhibitors further increases ropivacaine concentration. PMID- 11049911 TI - Characteristics of ropivacaine block of Na+ channels in rat dorsal root ganglion neurons. AB - When used for epidural anesthesia, ropivacaine can produce a satisfactory sensory block with a minor motor block. We investigated its effect on tetrodotoxin sensitive (TTX-S) and tetrodotoxin-resistant (TTX-R) Na(+) currents in rat dorsal root ganglion (DRG) neurons to elucidate the mechanisms underlying the above effects. Whole-cell patch-clamp recordings were made from enzymatically dissociated neurons from rat DRG. A TTX-S Na(+) current was recorded preferentially from large DRG neurons and a TTX-R Na(+) current preferentially from small ones. Ropivacaine shifted the activation curve for the TTX-R Na(+) channel in the depolarizing direction and the inactivation curve for both types of Na(+) channel in the hyperpolarizing direction. Ropivacaine blocked TTX-S and TTX-R Na(+) currents, but its half-maximum inhibitory concentration (IC(50)) was significantly lower for the latter current (116 +/- 35 vs 54 +/- 14 microM; P: < 0.01); similar IC(50) values were obtained with the (R)-isomer of ropivacaine. Ropivacaine produced a use-dependent block of both types of Na(+) channels. Ropivacaine preferentially blocks TTX-R Na(+) channels over TTX-S Na(+) channels. We conclude that because TTX-R Na(+) channels exist mainly in small DRG neurons (which are responsible for nociceptive sensation), such selective action of ropivacaine could underlie the differential block observed during epidural anesthesia with this drug. IMPLICATIONS: Whole-cell patch-clamp recordings of tetrodotoxin-sensitive and tetrodotoxin-resistant Na(+) currents in rat dorsal root ganglion neurons showed ropivacaine preferentially blocked tetrodotoxin resistant Na(+) channels over tetrodotoxin-sensitive Na(+) channels. This could provide a desirable differential sensory blockade during epidural anesthesia using ropivacaine. PMID- 11049912 TI - Postoperative analgesic effects of celecoxib or rofecoxib after spinal fusion surgery. AB - Nonsteroidal antiinflammatory drugs are recommended for the multimodal management of postoperative pain and may have a significant opioid-sparing effect after major surgery. The analgesic efficacy of the cyclooxygenase-2 nonsteroidal antiinflammatory drugs, celecoxib and rofecoxib, have not been evaluated after major orthopedic surgery. This study was designed to determine whether the administration of a preoperative dose of celecoxib or rofecoxib to patients who have undergone spinal stabilization would decrease patient-controlled analgesia (PCA) morphine use and/or enhance analgesia. We evaluated 60 inpatients undergoing spine stabilization by one surgeon. All patients received PCA morphine. The patients were divided into three groups. Preoperatively, they were given oral celecoxib 200 mg, rofecoxib 50 mg, or placebo. The outcome measures included pain scores and 24-h morphine use at six times during the first 24 postoperative h. The total dose of morphine and the cumulative doses for each of the six time periods were significantly more in the placebo group than in the other two groups. The morphine dose was significantly less in five of the six time intervals in the rofecoxib group compared with the celecoxib group. The pain scores were significantly less in the rofecoxib group than in the other two groups at two of the six intervals, and less than the placebo group in an additional interval. Although both rofecoxib and celecoxib produce similar analgesic effects in the first 4 h after surgery, rofecoxib demonstrated an extended analgesic effect that lasted throughout the 24-h study. We thus recommend that rofecoxib be used as a preoperative component of pain management that includes PCA morphine in patients undergoing spine stabilization surgery. IMPLICATIONS: The cyclooxygenase-2-specific nonsteroidal antiinflammatory drugs, celecoxib and rofecoxib, both demonstrate an opioid-sparing effect after spinal fusion surgery. Celecoxib resulted in decreased morphine use for the first 8 h after surgery, whereas rofecoxib demonstrated less morphine use throughout the 24 h study period. PMID- 11049913 TI - Patient-controlled analgesia with tramadol versus tramadol plus lysine acetyl salicylate. AB - By using a patient-controlled analgesia (PCA) delivery system, we compared the clinical advantages and disadvantages of PCA with tramadol and PCA with a mixture of tramadol plus lysine acetyl salicylate (a soluble aspirin). Fifty adult patients who had undergone major orthopedic surgeries were enrolled into a prospective, randomized, and double-blinded study. The general anesthesia was performed in a standard manner. At the beginning of wound closure, an equal volume dose of either tramadol 2.5 mg/kg (Group 1) or tramadol 1.25 mg/kg + lysine acetyl salicylate 12.5 mg/kg mixture (Group 2) was administered slowly IV. These solutions were continued postoperatively for IV PCA. Pain control, patient satisfaction, vital signs, and adverse effects were assessed for 48 h. Visual Analog Scale 2 h received either droperidol (0.625 mg IV) or a placebo before emergence. Patients requiring treatment for PONV in the postanesthesia care unit were randomized to receive either droperidol (0.625 mg IV), ondansetron (4 mg IV), or promethazine (12. 5 mg IV). Droperidol effectively prevented PONV (6.8% in droperidol-treated patients versus 40.8% in placebo-treated patients, P: < 0.001). Droperidol, ondansetron, and promethazine were equally effective in treating established PONV, without significant differences in side effects or time to postanesthesia care unit discharge. IMPLICATIONS: Droperidol 0.625 mg IV before emergence from general anesthesia effectively reduces postoperative nausea and vomiting (PONV) in the general surgical population. Our randomized, double blinded, placebo-controlled study demonstrated a reduction in PONV from 41% to 7%. Droperidol is a safe and inexpensive alternative to ondansetron. Droperidol, ondansetron, and promethazine are also equally effective in treating PONV in the postanesthesia care unit. PMID- 11049919 TI - The effects of prolonged low-flow sevoflurane anesthesia on renal and hepatic function. AB - We assessed the effects of prolonged low-flow sevoflurane anesthesia on renal and hepatic functions by comparing high-flow sevoflurane with low-flow isoflurane anesthesia. Thirty patients scheduled for surgery of > or =10 h in duration randomly received either low-flow (1 L/min) sevoflurane anesthesia (n = 10), high flow (6-10 L/min) sevoflurane anesthesia (n = 10), or low-flow (1 L/min) isoflurane anesthesia (n = 10). We measured the circuit concentrations of Compound A and serum fluoride. Renal function was assessed by blood urea nitrogen, serum creatinine, creatinine clearance, and urinary excretion of glucose, albumin, protein, and N:-acetyl-beta-D-glucosaminidase. The hepatic function was assessed by serum aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, alkaline phosphatase, and total bilirubin. Compound A exposure was 277 +/- 120 (135-478) ppm-h (mean +/- SD [range]) in the low-flow sevoflurane anesthesia. The maximum concentration of serum fluoride was 53.6 +/- 5.3 (43.4-59.3) micromol/L for the low-flow sevoflurane anesthesia, 47.1 +/- 21.2 (21.4-82.3) micromol/L for the high-flow sevoflurane anesthesia, and 7.4 +/- 3.2 (3.2-14.0) micromol/L for the low-flow isoflurane anesthesia. Blood urea nitrogen and serum creatinine were within the normal range, and creatinine clearance did not decrease throughout the study period in any group. Urinary excretion of glucose, albumin, protein, and N: acetyl-beta-D-glucosaminidase increased after anesthesia in all groups, but no significant differences were seen among the three groups at any time point after anesthesia. Lactate dehydrogenase and alkaline phosphatase on postanesthesia Day 1 were higher in the high-flow sevoflurane group than in the low-flow sevoflurane group. However, there were no significant differences in any other hepatic function tests among the groups. We conclude that prolonged low-flow sevoflurane anesthesia has the same effect on renal and hepatic functions as high-flow sevoflurane and low-flow isoflurane anesthesia. IMPLICATIONS: During low-flow sevoflurane anesthesia, intake of Compound A reached 277 +/- 120 ppm-h, but the effect on the kidney and the liver was the same in high-flow sevoflurane and low flow isoflurane anesthesia. PMID- 11049920 TI - A comparison of remifentanil and sufentanil as adjuvants during sevoflurane anesthesia with epidural analgesia for upper abdominal surgery: effects on postoperative recovery and respiratory function. AB - We compared the recovery profile and postoperative SpO(2) after the administration of general anesthesia with either sevoflurane-remifentanil or sevoflurane-sufentanil in 30 healthy patients undergoing upper abdominal surgery. They were randomly allocated to receive general anesthesia with sevoflurane and small doses of either remifentanil (n = 15) or sufentanil (n = 15), followed by postoperative epidural analgesia. The median sevoflurane minimum alveolar anesthetic concentration-hour was 2.3 (1.2-6.3) in group Remifentanil and 2.6 (1.4-5.2) in group Sufentanil (P: = 0.39), while the median consumption of remifentanil was 1.3 mg (0.7-3.4 mg) and sufentanil 0.09 mg (0.05-0.6 mg). Tracheal extubation required 10 min (6-18 min) with remifentanil and 14 min (8-24 min) with sufentanil (P: = 0.05); however, no differences in time to discharge from the recovery area were reported (24 min [12-75 min] with remifentanil and 30 min [12-135 min] with sufentanil; P: = 0. 35). From the first to seventh hour after surgery, SpO(2) was decreased more in the sufentanil than in the remifentanil group (P: = 0.001), and seven patients in the sufentanil group showed at least one episode with SpO(2) < or = 90% for more than 1 min (P: = 0.006) (median: 1 episode; range: 0-17 episodes; P: = 0.003). When added to sevoflurane, remifentanil is as effective as sufentanil during the intraoperative period, but provides shorter time to tracheal extubation and fewer effects on postoperative SpO(2) in the first 7 h after surgery. IMPLICATIONS: In this double blinded study, we evaluated the effects of adding small infusions of either remifentanil or sufentanil to sevoflurane in combination with postoperative epidural analgesia for upper abdominal surgery. We demonstrated that remifentanil is as effective as sufentanil during the intraoperative period, but that it provides shorter time to extubation and fewer effects on postoperative SpO(2) in the first 7 h after surgery. PMID- 11049921 TI - Cervical spine motion during airway management: a cinefluoroscopic study of the posteriorly destabilized third cervical vertebrae in human cadavers. AB - We conducted a randomized, controlled, crossover study to determine cervical spine motion for six airway management techniques in human cadavers with a posteriorly destabilized third cervical (C-3) vertebra. A destabilized C-3 segment was created in 10 cadavers (6-24 h postmortem). Cervical motion was recorded by continuous lateral fluoroscopy. The following airway management techniques were performed in random order on each cadaver with manual in-line stabilization applied: face mask ventilation (FM), laryngoscope-guided orotracheal intubation (OETT), fiberscope-guided nasal intubation (FOS-NETT), esophageal tracheal Combitube((R)) (Kendall-Sheridan, Neustadt, Germany) insertion (ETC), intubating laryngeal mask insertion with fiberscope-guided tracheal intubation (ILM-OETT), and laryngeal mask airway insertion (LMA). Afterward, maximum head-neck flexion (FLEX-MAX) and maximum head-neck extension (EXT-MAX) without manual in-line stabilization was performed to determine maximum motion. The maximum posterior displacement of C-3 and the maximum segmental sagittal motion of C2-3 were determined. There was a significant increase in posterior displacement for the FM (1.9 +/- 1.2 mm, P: < 0.01), OETT (2.6 +/- 1.6 mm, P: < 0.0001), ETC (3.2 +/- 1.6 mm, P: < 0.0001), ILM-OETT (1.7 +/- 1.3 mm, P: < 0. 01), LMA (1.7 +/- 1.3 mm, P: < 0.01), FLEX-MAX (3.7 +/- 1.9 mm, P: < 0.0001), EXT-MAX (1.8 +/- 1.7, P: < 0.01), however, not for FOS-NETT (0.1 +/- 0.7 mm). Posterior displacement was less for the ILM-OETT and LMA than for the ETC (both P: < 0.04). There were no significant increases in segmental sagittal motion with any airway manipulation other than with FLEX-MAX (-4.5 +/- 4.0 degrees, P: < 0.01). Posterior displacement was similar to FLEX-MAX for the OETT and ETC; however, it was less for the FM, FOS-NETT, ILM-OETT, and LMA (all P: < 0.01). Posterior displacement was similar to EXT-MAX for all airway manipulations other than for FOS-NETT (P: < 0.001). For cervical motion and the techniques tested, the safest method of airway management in a patient with a posteriorly destabilized C-3 segment is FOS-NETT. LMA devices may be preferable to the ETC. IMPLICATIONS: In the cadaver model of a destabilized third cervical vertebrae, significant displacement of the injured segment occurs during airway management with the face mask, laryngoscope-guided oral intubation, the esophageal tracheal Combitube (Kendall-Sheridan, Neustadt, Germany), the intubating and standard laryngeal mask airway; but not with fiberscope-guided nasal intubation. For cervical motion and the techniques tested, the safest airway technique with this injury is fiberscope-guided nasotracheal intubation. Laryngeal mask devices are preferable to the esophageal tracheal Combitube. PMID- 11049922 TI - Thromboelastography identifies sex-related differences in coagulation. AB - Thromboelastography is an in vitro, point-of-care monitor of whole blood coagulation. Thromboelastography studies have demonstrated a hypercoagulable state during pregnancy. Perhaps the hypercoagulability is attributable to female sex hormones. The aim of the study was to determine if sex, in addition to pregnancy, affected thromboelastography variables by studying male and female (pregnant and nonpregnant) volunteers. Thromboelastography showed significant (P:<0.01) differences in sex, with a significant (P: < 0. 0001) trend of increasing whole blood coagulability from men through nonpregnant to pregnant women. The thromboelastograph, used as a diagnostic tool, shows that women have more whole blood coagulability than men. IMPLICATIONS: The thromboelastograph, used as a diagnostic tool, shows that women have more whole blood coagulability than men. PMID- 11049923 TI - Isoflurane depresses electroencephalographic and medial thalamic responses to noxious stimulation via an indirect spinal action. AB - Anesthetics such as isoflurane act in the spinal cord to suppress movement in response to noxious stimulation. Spinal anesthesia decreases hypnotic/sedative requirements, possibly by decreasing afferent transmission of stimuli. We hypothesized that isoflurane action in the spinal cord would similarly depress the ascending transmission of noxious input to the thalamus and cerebral cortex. In six isoflurane-anesthetized goats, we measured electroencephalographic (EEG) and thalamic single-unit responses to a clamp applied to the forelimb. Cranial bypass permitted differential isoflurane delivery to the torso and cranial circulations. When the cranial-torso isoflurane combination was 1.3% +/- 0.2% 1.0% +/- 0.4% the noxious stimulus did not evoke significant changes in the EEG or thalamic activity: 389 (153-544) to 581 (172-726) impulses/min, (median, 25th 75th percentile range, P: > 0.05). When the cranial-torso isoflurane combination was 1.3% +/- 0.2%-0.3% +/- 0.2%, noxious stimulation increased thalamic activity: 804 (366-1162) to 1124 (766-1865) impulses/min (P: < 0.05), and the EEG "desynchronized": total EEG power decreased from 25 +/- 20 microV(2) to 12 +/- 8 microV(2) (P: < 0.05). When the cranial-torso isoflurane was 1.7% +/- 0.1%-0.3% +/- 0.2%, the noxious stimulus did not significantly affect thalamic: 576 (187 738) to 1031 (340-1442) impulses/min (P: > 0.05), or EEG activity. The indirect torso effect of isoflurane on evoked EEG total power (12.6 +/- 2.7 microV(2)/vol%, mean +/- SE) was quantitatively similar to the direct cranial effect (17.7 +/- 3.0 microV(2)/vol%; P: > 0.05). These data suggest that isoflurane acts in the spinal cord to blunt the transmission of noxious inputs to the thalamus and cerebral cortex, and thus might indirectly contribute to anesthetic endpoints such as amnesia and unconsciousness. IMPLICATIONS: Isoflurane action in the spinal cord diminished the transmission of noxious input to the brain. Because memory and consciousness are likely dependent on the "arousal" state of the brain, this indirect action of isoflurane could contribute to anesthetic-induced amnesia and unconsciousness. PMID- 11049924 TI - The anesthetic potency of propanol and butanol versus propanethiol and butanethiol in alpha1 wild type and alpha1(S267Q) glycine receptors. AB - Although similar in shape and size, and although differing only by substitution of a sulfur atom for an oxygen atom, propanethiol and butanethiol differ markedly from propanol and butanol in their in vivo potency and physical properties. Recent theories of narcosis suggest that anesthetics may act by enhancing the effect of inhibitory agonists, such as glycine, on their receptors. We tested whether propanol, butanol, propanethiol, and butanethiol enhance the effect of glycine on alpha1 glycine receptors expressed in Xenopus laevis oocytes in a manner that reflects the in vivo differences found for potencies. As anticipated, we found an immediate parallel between in vivo (rat minimum alveolar concentration of anesthetic required to eliminate movement in response to a noxious stimulus in 50% of subjects) and in vitro (recombinant receptor) effects. All four compounds enhanced the effect of glycine on wild type receptors, and the extent of enhancement for a given minimum alveolar concentration-multiple was approximately the same for all compounds. We also found that propanethiol, butanethiol, propanol, and butanol did not affect, or minimally affected, the action of glycine in anesthetic resistant mutants in which the amino acid serine at position 267 was replaced by glutamine [alpha1(S267Q)]. IMPLICATIONS: The in vivo potencies of propanethiol, butanethiol, propanol, and butanol correlate with their capacities to enhance the effect of glycine on alpha1 glycine receptors expressed in Xenopus laevis oocytes. These results support the notion that a protein mediates anesthetic action. PMID- 11049925 TI - The anesthetic potencies of alkanethiols for rats: relevance to theories of narcosis. AB - Meyer and Overton suggested that anesthetic potency correlates inversely with lipophilicity. Thus, MAC times the olive oil/gas partition coefficient equals an approximately constant value of 1.82 +/- 0.56 atm (mean +/- SD). MAC is the minimum alveolar concentration of anesthetic required to eliminate movement in response to a noxious stimulus in 50% of subjects. Although MAC times the olive oil/gas partition coefficient also equals an approximately constant value for normal alkanols from methanol through octanol, the value (0.156 +/- 0.072 atm) is 1/10th that found for conventional anesthetics. We hypothesized that substitution of sulfur for the oxygen in n-alkanols would decrease their saline/gas partition coefficients (i.e., decrease polarity) while sustaining lipid/gas partition coefficients. Further, we hypothesized that these changes would produce products of MAC times olive oil partition coefficients that approximate those of conventional anesthetics. To test these predictions, we measured MAC in rats, and saline and olive oil solubilities for the series H(CH(2))(n)SH, comparing the results with the series H(CH(2))(n)OH for compounds having three to six carbon atoms. As hypothesized, the alkanethiols had similar oil/gas partition coefficients, 1000-fold smaller saline gas partition coefficients, and MAC values 30 times greater than for comparable alkanols. Such findings are consistent with the notion that the greater potency of many alkanols (greater than would be predicted from conventional inhaled anesthetics and the Meyer-Overton hypothesis) results from their greater polarity. IMPLICATIONS: The in vivo anesthetic potency of alkanols and alkanethiols correlates with their lipophilicity and hydrophilicity. PMID- 11049926 TI - Urgent colectomy in a patient with membranous tracheal disruption after severe vomiting. AB - IMPLICATIONS: We report a case of a patient who developed membranous tracheal disruption after severe vomiting. He subsequently required urgent colectomy for toxic megacolon under general anesthesia. With this challenging situation, we were able to successfully conduct general anesthesia in the presence of tracheal laceration, pneumothorax, and pneumomediastinum. PMID- 11049927 TI - Reinforcement of laryngeal mask airway cuff position with endotracheal tube cuff for airway control in a patient with altered upper airway anatomy. AB - IMPLICATIONS: This case report suggests that the laryngeal mask airway (LMA) cuff position may not be optimal in some difficult airway situations in which the anatomical position of the larynx is altered. Reinforcement of the LMA cuff position by an additional cuff on the dorsal side of the LMA cuff may prove helpful. In this case, in which a difficult airway was anticipated, a nasopharyngeal tube cuff placed behind the standard LMA cuff helped relieve upper airway obstruction. PMID- 11049928 TI - Peripheral nerve stimulators for regional anesthesia can generate excessive voltage output with poor ground connection. PMID- 11049930 TI - Sister Maria's test to differentiate between cerebrospinal fluid and a local anesthetic. PMID- 11049929 TI - N(2)O usage in laparoscopic cases. PMID- 11049931 TI - Any propofol compatibility study must include an emulsion stability analysis. PMID- 11049932 TI - Coffee is not a clear liquid. PMID- 11049933 TI - New jaw support device and awake fiberoptic intubation. PMID- 11049934 TI - Foreign body removal: tracheal backflow air or rigid bronchoscopy? PMID- 11049935 TI - Ropivacaine and bupivacaine with fentanyl for labor epidural anesthesia. PMID- 11049936 TI - Are Medicare patients sicker, more complex, and at a higher risk for perioperative complications? PMID- 11049937 TI - Are postdural puncture symptoms immediate in elderly patients? PMID- 11049938 TI - The cost effectiveness of anesthesia workforce models: a critique. PMID- 11049939 TI - The cost effectiveness of anesthesia workforce models: the creation of a procrustean bed. PMID- 11049940 TI - The continuing need to publish laboratory science in clinical journals. PMID- 11049941 TI - From eye spots to eye shine. PMID- 11049942 TI - Diabetes and primary open angle glaucoma. PMID- 11049943 TI - Glaucoma incidence in an unselected cohort of diabetic patients: is diabetes mellitus a risk factor for glaucoma? DARTS/MEMO collaboration. Diabetes Audit and Research in Tayside Study. Medicines Monitoring Unit. AB - AIMS: To evaluate whether diabetes mellitus is a risk factor for the development of primary open angle glaucoma or ocular hypertension (OHT). METHODS: A historical cohort study of an unselected population comprising all residents of the Tayside region of Scotland was performed using record linkage techniques followed by case note review. Ascertainment of prevalent diabetes was achieved using the Diabetes Audit and Research in Tayside Study (DARTS) validated regional diabetes register. Glaucoma and treated OHT were defined by encashment of community prescriptions and the statutory surgical procedure coding database. RESULTS: The study population comprised 6631 diabetic subjects and 166 144 non diabetic subjects aged >40 years without glaucoma or OHT at study entry. 65 patients with diabetes and 958 without diabetes were identified as new cases of glaucoma or treated OHT during the 24 month study period, yielding a standardised morbidity ratio of 127 (95% CI, 96-158). Case note review demonstrated non differential misclassification of prevalent glaucoma and OHT as incident disease (diabetic cohort 20%, non-diabetic cohort 24%; p=0.56) primarily as a result of non-compliance in medically treated disease. Removing misclassified cases and adjusting for age yielded an incidence of primary open angle glaucoma in diabetes of 1.1/1000 patient years (95% CI, 0.89-1. 31) compared to 0.7/1000 patient years (95% CI, 0.54-0.86) in the non-diabetic cohort; RR 1.57 (95% CI, 0.99-2.48). CONCLUSIONS: This study failed to confirm an association between diabetes mellitus and primary open angle glaucoma and ocular hypertension. A non significant increase in diagnosed and treated disease in the diabetic population was observed, but evidence was also found that detection bias contributes to this association. PMID- 11049944 TI - Impression cytology of the conjunctival epithelium in patients with chronic renal failure. AB - AIMS: To assess ocular surface changes in patients with chronic renal failure (CRF), to compare the results with the degree of corneo-conjunctival calcium deposits, and to determine whether precipitation of calcium salts predisposes ocular surface modifications. METHODS: Impression cytology from 50 CRF patients on regular haemodialysis and 22 age and sex matched control subjects were studied. Specimens were obtained from the temporal bulbar conjunctiva using cellulose acetate filter paper. The samples were fixed in 95% ethanol, stained with the periodic acid Schiff (PAS) stain, and evaluated by light microscopy and were graded by a masked observer. Corneo-conjunctival calcification was graded by the Porter and Crombie classification. RESULTS: In the study group, three patients (6%) disclosed grade 0, 14 patients (28%) grade 1, and 33 patients (66%) grade 2-3 cytological changes. There was a statistically significant difference between the patient and the control groups (p= 0.0007), but no correlation could be found between the impression cytology grades and the calcium deposit grades (p=0.62). CONCLUSION: The ocular surfaces of CRF patients differ significantly from those of normal individuals, and it can be detected using impression cytology. These data suggest that the severity of conjunctival changes are not related to the presence or extent of calcium deposition. PMID- 11049945 TI - Anti-inflammatory and antiallergic effects of ketorolac tromethamine in the conjunctival provocation model. AB - AIM: To study the effect of the topical anti-inflammatory drug, ketorolac, on (1) the clinical allergic reaction induced by the conjunctival provocation test (CPT); (2) the release of tryptase in tears; and (3) the expression of adhesion molecules on the conjunctival epithelium. METHODS: 10 allergic but non-active patients were challenged in both eyes with increasing doses of specific allergen to obtain a positive bilateral reaction and rechallenged, after 1 week, to confirm the allergic threshold dose response. After 2 weeks, a third CPT was then performed bilaterally 30 minutes after topical application of ketorolac in one eye and placebo in the contralateral eye in a double blind fashion. Clinical symptoms and signs were registered 5, 10, 15, and 20 minutes after challenge. The following objective tests were performed: tear tryptase measurement; tear cytology; and conjunctival impression cytology for immunohistochemical expression of ICAM-1 on epithelial cells. RESULTS: Compared with placebo, ketorolac significantly reduced the total clinical score and the itching score in the 20 minutes after challenge (p<0.0005). Tear levels of tryptase were significantly reduced in the ketorolac pretreated eyes compared with placebo (p<0.03). Eosinophils, neutrophils, and lymphocytes in tear cytology were significantly lower in ketorolac treated eyes compared with placebo. A significant difference in the epithelial expression of ICAM-1 was observed between placebo and ketorolac treated eyes (p<0.05). CONCLUSION: Ketorolac proved to be effective in reducing mast cell degranulation, as indicated by significantly decreased tryptase tear levels, as well as the clinical and cytological allergic reaction. PMID- 11049946 TI - Central corneal thickness determined with optical coherence tomography in various types of glaucoma. AB - AIMS: To evaluate central corneal thickness determined by optical coherence tomography (OCT) in various types of glaucoma, and its influence on intraocular pressure (IOP) measurement. METHODS: Central corneal thickness (CCT) was determined by using OCT in 167 subjects (167 eyes). 20 had primary open angle glaucoma (POAG), 42 had low tension glaucoma (LTG), 22 had ocular hypertension (OHT), 10 had primary angle closure glaucoma (AC), 24 had pseudoexfoliation glaucoma (PEX), 13 had pigmentary glaucoma (PIG), and 36 were normal. RESULTS: CCT was significantly higher in ocular hypertensive subjects (593 (SD 35) microm, p <0.0001) than in the controls (530 (32) microm), whereas patients with LTG (482 (28) microm, p < 0. 0001), PEX (493 (33) microm, p <0.0001), and POAG (512 (30) microm, p <0.05) showed significantly lower readings. There was no statistically significant difference between the controls and patients with PIG (510 (39) microm) and AC (539 (37) microm). CONCLUSIONS: Because of thinner CCT in patients with LTG, PEX, and POAG this may result in underestimation of IOP, whereas thicker corneas may lead to an overestimation of IOP in subjects with OH. By determining CCT with OCT, a new and precise technique to measure CCT, this study emphasises the need for a combined measurement of IOP and CCT in order to obtain exact IOP readings. PMID- 11049947 TI - Detection of herpes simplex virus type 1, 2 and varicella zoster virus DNA in recipient corneal buttons. AB - AIM: To study the value of polymerase chain reaction (PCR) analysis, to detect viral DNA in recipient corneal buttons taken at the time of penetrating keratoplasty (PKP) in patients with an initial diagnosis of herpetic stromal keratitis (HSK). Since HSK has a tendency to recur, an accurate diagnosis of previous HSK could be the reason to start antiviral treatment immediately, thereby possibly decreasing the number of graft failures due to recurrent herpetic keratitis. METHODS: Recipient corneal buttons and aqueous humour (AH) samples were obtained at the time of PKP from HSK patients (n=31) and from other patients (n=78). Eye bank corneas were also used (n=23). Herpes simplex virus type 1 (HSV-1), type 2 (HSV-2), and varicella zoster virus (VZV) infection were assessed by PCR and antibody detection. RESULTS: The clinical diagnosis HSK could be confirmed by PCR for HSV-1 in 10/31 (32%). In these corneal buttons HSV-2 DNA was detected in 1/31 (3%) and VZV DNA in 6/31 (19%). Intraocular anti-HSV antibody production was detected in 9/28 AH samples tested (32%). In the other patient derived corneas HSV-1 DNA was detected in 13/78 (17%), including eight failed corneal grafts without clinically obvious herpetic keratitis in the medical history. In clear eye bank corneas HSV-1 was detected in 1/23 (4%). CONCLUSIONS: PCR of HSV-1 on corneal buttons can be a useful diagnostic tool in addition to detection of intraocular anti-HSV antibody production. Furthermore, the results were suggestive for the involvement of corneal HSV infection during allograft failure of corneas without previous clinical characteristic signs of herpetic keratitis. PMID- 11049948 TI - Mooren's ulcer in China: a study of clinical characteristics and treatment. AB - AIMS: To investigate the clinical characteristics and compare the effects of several methods of treatment of Mooren's corneal ulcer. METHODS: 550 consecutive cases of Mooren's corneal ulcer were analysed in patients, including age, sex, laterality of eye, ulcer location, perforative rate, cure rate of surgeries, recurrent rate, the effects of conjunctiva excision, lamellar keratoplasty (LKP), and LKP plus 1% cyclosporin A eye drops. RESULTS: The average age of onset was 48.4 years of age. The ratio of males to females was 1:0. 74. 165 (30%) cases had the disease bilaterally, of which 52 (31.5%) occurred in the young age group and 113 (68.5%) in the old age group. Ulcers of 501 eyes (70.1%) were located at the limbus of the palpebral fissure. The perforation rate was 13.3%, with perforation of 41 eyes (43.2%) occurring in the young age group and 54 (56.8%) in the old age group. Postoperative recurrence rate was 25.6%. The cure rate of the first procedure of LKP plus 1% cyclosporin A eye drops was 73.7%. The final cure rate was 95.6%, and the postoperative preservation rate of the eye globe was 99.7%. CONCLUSION: This primary study provided the clinical characteristics of patients with Mooren's corneal ulcer in China. LKP plus 1% cyclosporin A eye drops was an effective treatment. PMID- 11049949 TI - Changes in ocular surface caused by antiglaucomatous eyedrops: prospective, randomised study for the comparison of 0.5% timolol v 0. 12% unoprostone. AB - AIM: To study changes induced in ocular surface epithelia and the tear film by antiglaucomatous eyedrops. A beta blocker (0.5% timolol) and a novel prostaglandin F(2alpha) metabolite related drug (0.12% unoprostone) were examined in a prospective, randomised fashion. METHODS: 40 patients were randomly assigned to use either 0. 5% timolol (timolol group) or 0.12% unoprostone eyedrops (unoprostone group) twice a day for 24 weeks. In addition to routine ocular examinations, corneal epithelial integrity (vital staining tests, tear film break up time (BUT), anterior fluorometry, specular microscopy) and tear function (Schirmer's test, cotton thread test, tear clearance test (TCT)) were examined before and after the treatment. RESULTS: Both eyedrops caused significant reduction in intraocular pressure from the baseline levels. No significant changes were noted in corneal integrity in both groups, except a decrease in BUT at 20 weeks in the timolol group. The timolol group demonstrated significant decreases in Schirmer's test, tear clearance test, and tear function index (Schirmer's test value multiplied by clearance test); however, no such changes were noted in the unoprostone group. CONCLUSION: While unoprostone eyedrops caused no adverse effects on the corneal epithelial integrity and tear function, timolol caused significant impairments in tear production and turnover. PMID- 11049950 TI - YAG laser iridotomy treatment for primary angle closure in east Asian eyes. AB - AIM: To assess the efficacy of Nd:YAG laser iridotomy as initial treatment for primary angle closure in a community setting in rural Mongolia. METHODS: Subjects with occludable drainage angles in two glaucoma prevalence surveys in Mongolia (carried out in 1995 and 1997) were treated with YAG laser iridotomy at the time of diagnosis. These patients were re-examined in 1998. Patency of iridotomy, intraocular pressure (IOP), visual acuity, and gonioscopic findings were recorded. Iridotomy was classified unsuccessful in eyes where further surgical intervention was required or in which there was a loss of visual acuity to <3/60 from glaucomatous optic neuropathy. RESULTS: 164 eyes of 98 subjects were examined. Patent peripheral iridotomies were found in 98.1% (157/160) of eyes that had not undergone surgery. Median angle width increased by two Shaffer grades following iridotomy. Iridotomy alone failed in 3% eyes with narrow drainage angles and either peripheral anterior synechiae or raised IOP, but normal optic discs and visual fields. However, in eyes with established glaucomatous optic neuropathy at diagnosis iridotomy failed in 47%. None of the eyes with occludable angles that were normal in all other respects, and underwent iridotomy, developed glaucomatous optic neuropathy or symptomatic angle closure within the follow up period. CONCLUSIONS: Nd: YAG laser iridotomy is effective in widening the drainage angle and reducing elevated IOP in east Asian people with primary angle closure. This suggests that pupil block is a significant mechanism causing closure of the angle in this population. Once glaucomatous optic neuropathy associated with synechial angle closure has occurred, iridotomy alone is less effective at controlling IOP. PMID- 11049951 TI - Local anaesthetic techniques and pulsatile ocular blood flow. AB - AIM: To compare pulsatile ocular blood flow (POBF) and intraocular pressure (IOP) between eyes of patients receiving either peribulbar (with and without balloon compression) or subconjunctival local anaesthesia (LA). METHODS: 30 eyes of 30 patients undergoing cataract surgery by phacoemulsification were investigated in a study of parallel group design. Ten patients had peribulbar LA and 10 minutes compression with a Honan's balloon (group A). A further 10 patients who received peribulbar LA alone (group B) acted as controls for the effects of balloon compression. Ten other patients were given subconjunctival LA (group C). POBF and IOP were measured using a modified Langham pneumatonometer. Three measurements were made in each eye, the first recording immediately before LA, the second 1 minute after, and the third 10 minutes after LA. RESULTS: No significant change in POBF or IOP was recorded in eyes receiving subconjunctival LA. In the peribulbar groups (A and B), there was a drop in median POBF of 252 and 138 microl/min respectively 1 minute after LA, which was statistically significant in both groups (p<0. 01). By 10 minutes, POBF tended to return to baseline levels, but remained significantly reduced in group B (p<0.05). In addition, there was a significant (p<0.05) reduction in IOP (mean drop of 4.82 mm Hg) in group A following peribulbar LA with balloon compression. CONCLUSIONS: POBF was significantly reduced after peribulbar LA but was unchanged after subconjunctival LA. Balloon compression reduced IOP and improved POBF following peribulbar LA. The findings may have clinical implications in patients with compromised ocular circulation or significant glaucomatous optic neuropathy. PMID- 11049952 TI - Acute posterior vitreous detachment: the predictive value of vitreous pigment and symptomatology. AB - AIM: To establish whether the presence of a retinal break can be predicted either by the presence of a positive Shafer's sign (pigment granules in the anterior vitreous) or symptomatology in patients presenting with an acute posterior vitreous detachment (PVD). METHODS: 200 eyes of 200 phakic patients with a symptomatic PVD of less than 1 month's duration underwent documentation of symptomatology and examination of the anterior vitreous for the presence of pigment granules. Indentation ophthalmoscopy was then carried out by an experienced vitreoretinal surgeon with no knowledge of the symptomatology or anterior vitreous gel examination findings. A second prospective group of 115 consecutive patients were assessed in a similar manner before primary rhegmatogenous retinal detachment repair. RESULTS: In 200 eyes presenting with an acute PVD, 25 were found to have an associated retinal break, 23 of which were also Shafer positive. In 115 eyes presenting for retinal detachment repair, 111 had an associated PVD and were found to be Shafer positive. Symptomatology was not predictive of an associated retinal break in the PVD group or in those presenting with a retinal detachment. CONCLUSION: The increased use of Shafer's sign is recommended as a valuable aid in determining which patients require urgent referral for an expert retinal examination. It is not possible to predict those patients with a retinal break secondary to PVD on the basis of symptomatology alone. PMID- 11049953 TI - Clinicopathological correlation of deep retinal vascular anomalous complex in age related macular degeneration. AB - AIMS: To analyse the histopathology of "deep retinal vascular anomalous complex" or "chorioretinal anastomosis". METHODS: Six patients with a deep retinal vascular anomalous complex (age range 66-88 years) had fundus photography and fluorescein angiography not more than 14 days before foveal translocation surgery. Four patients were also documented with indocyanine green angiography. The surgical specimens were serially sectioned and stained in a stepped fashion with Masson trichrome, periodic acid Schiff, and phosphotungstic acid haematoxylin, a histochemical stain for fibrin. RESULTS: A subretinal fibrovascular membrane was surrounded by a rim consisting of diffuse drusen (basal laminar deposits), retinal pigment epithelium, and amorphous, fibrinous material interspersed with remains of outer segments in all specimens. In two specimens vascular structures were identified that left the specimen towards the retina. Amorphous material with the remains of outer segments was not found on the retinal side of the fibrovascular tissue itself but in four specimens a small neuroretinal portion (outer nuclear layer) was adherent to the complex. In three specimens a thin fibrocellular membrane was seen at the choroidal side of the diffuse drusen. CONCLUSION: Deep retinal vascular anomalous complex represents histologically neovascularisation growing out of the neuroretina, into the subretinal space, which mimics choroidal neovascularisation. The term therefore appears rightly chosen. PMID- 11049954 TI - Postmortem histological survey of the ocular lesions in a British population of AIDS patients. AB - AIMS: To study ocular pathology and systemic correlations in a series of 73 postmortem eyes from British patients who died from AIDS before the introduction of a HAART regimen. METHODS: The eyes were studied with conventional histology, special histochemical stainings, and immunohistochemistry. RESULTS: 72.6% of the cases showed chronic uveal inflammation, caused by opportunistic agents in 37.7% of them (cytomegalovirus (CMV) in 30.1%, C neoformans in 5.6%, and Gram positive bacteria in 1.8%). Cytoid bodies were noted in 10/73 eyes, three linked to CMV retinitis. Six retinal haemorrhages, four of which were secondary to CMV, were found. 14 specimens (19. 1%) showed foci of calcification, and a further 11 (15%) calcium oxalate deposits. In no cases were the calcific deposits suspected clinically. Six eyes (8.2%) did not show any abnormality. CONCLUSIONS: CMV retinitis is the most common (28.7%, 21/73) ocular infection in this series and may occur either during or in the absence of systemic dissemination. Conversely, ocular cryptococcosis appears to be an epiphenomenon of systemic and CNS disease. No other opportunistic ocular infections were present in this series. Interesting findings were the presence of intraocular precipitates of calcium oxalate and calcium phosphate or carbonate in a significant number of cases (15% and 19%, respectively), and the high prevalence of idiopathic uveal inflammation (43.8%). PMID- 11049955 TI - Ocular pulse amplitude in diabetes mellitus. AB - BACKGROUND: The influence of diabetes mellitus on ocular pulse amplitude (OPA), an indirect measure of choroidal perfusion, is unclear. METHODS: OPA, using the Langham ocular blood flow (OBF) system, applanation intraocular pressure (IOP), systemic blood pressure (BP), heart rate, and haemoglobin (Hb) A(1c) were measured in patients with insulin dependent diabetes mellitus (IDDM) with no (DR 0, n = 22) non-proliferative (DR-1, n = 24), and proliferative (DR-2, n = 18) diabetic retinopathy. RESULTS: Neither local (OPA, IOP) nor systemic perfusion parameters (BPs, HR) nor HbA(1c) were significantly altered in DR-0, DR-1, or DR 2 IDDM patients. CONCLUSION: Choroidal circulation remains unaffected as diabetic retinopathy advances. PMID- 11049956 TI - Evaluation of the Zeiss retinal vessel analyser. AB - AIM: To investigate the reproducibility and sensitivity of the Zeiss retinal vessel analyser, a new method for the online determination of retinal vessel diameters in healthy subjects. METHODS: Two model drugs were administered, a peripheral vasoconstrictor (the alpha receptor agonist phenylephrine) and a peripheral vasodilator (the nitric oxide donor sodium nitroprusside) in stepwise increasing doses. Nine healthy young subjects were studied in a placebo controlled double masked three way crossover design. Subjects received intravenous infusions of either placebo or stepwise increasing doses of phenylephrine (0.5, 1, or 2 microg/kg/min) or sodium nitroprusside (0.5, 1, or 2 microg/kg/min). Retinal vessel diameters were measured with the new Zeiss retinal vessel analyser. Retinal leucocyte velocity, flow, and density were measured with the blue field entoptic technique. The reproducibility of measurements was assessed with coefficients of variation and intraclass correlation coefficients. RESULTS: Placebo and phenylephrine did not influence retinal haemodynamics, although the alpha receptor antagonist significantly increased blood pressure. Sodium nitroprusside induced a significant increase in retinal venous and arterial diameters (p<0.001 each), leucocyte density (p=0.001), and leucocyte flow (p=0.024) despite lowering blood pressure to a significant degree. For venous and arterial vessel size measurements short term coefficients of variation were 1.3% and 2.6% and intraclass correlation coefficients were 0.98 and 0.96, respectively. The sensitivity was between 3% and 5% for retinal veins and 5% and 7% for retinal arteries. CONCLUSIONS: These data indicate that the Zeiss retinal vessel analyser is an accurate system for the assessment of retinal diameters in healthy subjects. In addition, nitric oxide appears to have a strong influence on retinal vascular tone. PMID- 11049958 TI - Ocular changes in heredo-oto-ophthalmo-encephalopathy. AB - BACKGROUND: Heredo-oto-ophthalmo-encephalopathy (HOOE) is a dominantly inherited disease characterised by gradual loss of vision from the age of 20, progressive hearing loss from the late 20s, cerebellar ataxia in the 30s, and death in dementia in the fourth or fifth decade. Currently, no detailed description has been given of the ocular changes seen in HOOE. Therefore, the ocular changes of HOOE were described on the basis of clinical and histological data from six affected family members. METHODS: Three members of the family affected by HOOE were subjected to a full ophthalmological re-examination, and postmortem examination was done on three eyes from two affected family members. RESULTS: Visual loss in HOOE was caused by posterior subcapsular cataract and retinal neovascularizations leading to vitreous haemorrhages and neovascular glaucoma. In the retina there was extensive accumulation of an amyloid material, both diffusely and in the walls of the retinal vessels. The retinal glial cells showed extensive pathological changes and retinal Muller cells were seen to occlude the lumen of retinal vessels. CONCLUSION: Heredo-oto-ophthalmo-encephalopathy is a familial amyloidosis of the central nervous system which is different from previously reported cases of amyloidosis by including cataract and retinal neovascularizations. The disease is accompanied by extensive changes in retinal glial cells that may play a part in the pathophysiology of the ocular complications of the disease. PMID- 11049957 TI - Prevalence of eye diseases in primary school children in a rural area of Tanzania. AB - AIMS: The study measured the prevalence of eye diseases in primary school children between 7 and 19 years of age in a rural area of Tanzania, and investigated whether teachers could successfully provide the first component of a school eye screening service. METHODS: Teachers from each of three primary schools in Mwanza Region tested visual acuity using a Snellen's E chart in 1438 pupils. 1386 of these pupils were then interviewed and underwent a full eye examination by an eye team. RESULTS: 10 pupils (0.7%) had bilateral poor eyesight (visual acuity worse than 6/12), and an additional 14 pupils (1.0%) had unilateral poor eyesight. Significant refractive errors causing visual acuity less than 6/12 (1.0%), strabismus (0. 5%), and amblyopia (0.2%) were uncommon. Overall, 76 pupils (5.5%) had active trachoma, though the prevalence was 15.5% in the poorest school. 73 pupils (5.3%) reported night blindness, eight (0.6%) had Bitot's spots, and 11 (0.8%) had corneal scars. Simple screening by teachers correctly identified 80% of the pupils who were found to have bilateral poor eyesight by the eye team, with 91% specificity. CONCLUSION: The prevalence of significant refractive errors was not high enough to justify a school eye screening programme solely for this purpose. However, a programme may be justified in areas where trachoma is common. Further research is needed to validate the frequent reports of night blindness and to establish the public health importance of vitamin A deficiency in this age group. PMID- 11049959 TI - Report of a family with dominantly inherited upper lid entropion. AB - AIM: To report the occurrence of late onset, bilateral, idiopathic upper lid entropion, occurring in three members of the same family, with a known family history. METHODS: Five family members were examined, and a history taken, at Moorfields Eye Hospital. Three patients were treated surgically, and one also had a tarsoconjunctival biopsy. RESULTS: In all cases, no aetiology was found. The family history suggests an autosomal dominant inheritance pattern. All patients were treated with anterior lamellar repositioning, and had optimal results. CONCLUSION: The family reported seems to be affected by a familial form of primary acquired upper lid entropion, that shows an autosomal dominant inheritance pattern. PMID- 11049960 TI - Enhancement of dedifferentiation and myoid differentiation of retinal pigment epithelial cells by platelet derived growth factor. AB - AIMS: To clarify factor(s) involved in morphological dedifferentiation of retinal pigment epithelial (RPE) cells in vitro from mitotically quiescent hexagonal cells to flattened cells that lack epithelial characteristics and concurrent myoid differentiation. METHODS: RPE cells which retained their differentiated hexagonal morphology were isolated from bovine eyes by mechanical pipetting. Dedifferentiation and myoid differentiation of RPE cells were examined by microscopic observation and immunohistochemical analysis using antibodies against cytokeratin, an epithelial marker, and alpha smooth muscle actin, a marker of myoid differentiation. The contractile ability of RPE cells was evaluated by collagen gel contraction assay. RESULTS: Platelet derived growth factor (PDGF) enhanced morphological changes in the RPE from hexagonal-shaped cells to flattened cells. Coincident with this morphological alteration, the expression of cytokeratin in RPE cells decreased and expression of alpha smooth muscle actin began and was increased in a time dependent manner. These alterations were completely blocked by collagen synthesis inhibitors. Interleukin 1beta, transforming growth factor beta1, insulin-like growth factor I, and basic fibroblast growth factor had little or no effect on the dedifferentiation. PDGF also potentiated the RPE induced collagen gel contraction. CONCLUSIONS: These results demonstrate that PDGF enhanced the dedifferentiation of RPE cells, the initial step of proliferative vitreoretinopathy (PVR), as well as myoid differentiation and collagen gel contraction. PDGF may have a versatile role in the pathogenesis of PVR involving collagen synthesis. PMID- 11049961 TI - Fibrillin and the eye. PMID- 11049963 TI - Current oculomotor research: physiological and psychological aspects PMID- 11049962 TI - Effect of spectacles on changes of spherical hypermetropia in infants. PMID- 11049964 TI - Glaucoma medical therapy. Principles and management PMID- 11049966 TI - Enucleation, evisceration and exenteration of the Eye PMID- 11049965 TI - Clinical strabismus management: principles and surgical techniques PMID- 11049967 TI - CD40-ligand (CD154) gene therapy for chronic lymphocytic leukemia. AB - Chronic lymphocytic leukemia (CLL) cells can be made to express recombinant CD40 ligand (CD154) by transduction with a replication-defective adenovirus vector (Ad CD154). Ad-CD154-transduced and bystander leukemia cells become highly effective antigen-presenting cells that can induce CLL-specific autologous cytotoxic T lymphocytes in vitro. This study investigated the immunologic and clinical responses to infusion of autologous Ad-CD154-CLL cells in patients with CLL. After a one-time bolus infusion of autologous Ad-CD154-transduced leukemia cells, there was increased or de novo expression of immune accessory molecules on bystander, noninfected CLL cells in vivo. Treated patients also developed high plasma levels of interleukin-12 and interferon-gamma, the magnitudes of which corresponded to absolute blood CD4(+) T-cell counts before therapy. On average, patients experienced a greater than 240% increase in absolute blood T-cell counts within 1 to 4 weeks of treatment. Moreover, treatment increased the numbers of leukemia-specific T cells, demonstrated by autologous ELISPOT assay and mixed lymphocyte reactions. These biologic effects were associated with reductions in leukemia cell counts and lymph node size. Treatment did not induce autoimmune thrombocytopenia or hemolytic anemia and no dose-limiting toxicity was observed. This approach may provide a novel and effective form of gene therapy for patients with this disease. PMID- 11049968 TI - Crystallographic structure and functional interpretation of the cytoplasmic domain of erythrocyte membrane band 3. AB - The red blood cell membrane (RBCM) is a primary model for animal cell plasma membranes. One of its major organizing centers is the cytoplasmic domain of band 3 (cdb3), which links multiple proteins to the membrane. Included among its peripheral protein ligands are ankyrin (the major bridge to the spectrin-actin skeleton), protein 4. 1, protein 4.2, aldolase, glyceraldehyde-3-phosphate dehydrogenase, phosphofructokinase, deoxyhemoglobin, p72syk protein tyrosine kinase, and hemichromes. The crystal structure of cdb3 is reported at 0.26 nm (2.6 A) resolution. A tight symmetric dimer is formed by cdb3; it is stabilized by interlocked dimerization arms contributed by both monomers. Each subunit also includes a larger peripheral protein binding domain with an alpha(+) beta-fold. The binding sites of several peripheral proteins are localized in the structure, and the nature of the major conformational change that regulates membrane skeletal interactions is evaluated. An improved structural definition of the protein network at the inner surface of the RBCM is now possible. PMID- 11049969 TI - A phase I/II trial of iodine-131-tositumomab (anti-CD20), etoposide, cyclophosphamide, and autologous stem cell transplantation for relapsed B-cell lymphomas. AB - Relapsed B-cell lymphomas are incurable with conventional chemotherapy and radiation therapy, although a fraction of patients can be cured with high-dose chemoradiotherapy and autologous stem-cell transplantation (ASCT). We conducted a phase I/II trial to estimate the maximum tolerated dose (MTD) of iodine 131 ((131)I)-tositumomab (anti-CD20 antibody) that could be combined with etoposide and cyclophosphamide followed by ASCT in patients with relapsed B-cell lymphomas. Fifty-two patients received a trace-labeled infusion of 1.7 mg/kg (131)I tositumomab (185-370 MBq) followed by serial quantitative gamma-camera imaging and estimation of absorbed doses of radiation to tumor sites and normal organs. Ten days later, patients received a therapeutic infusion of 1.7 mg/kg tositumomab labeled with an amount of (131)I calculated to deliver the target dose of radiation (20-27 Gy) to critical normal organs (liver, kidneys, and lungs). Patients were maintained in radiation isolation until their total-body radioactivity was less than 0.07 mSv/h at 1 m. They were then given etoposide and cyclophosphamide followed by ASCT. The MTD of (131)I-tositumomab that could be safely combined with 60 mg/kg etoposide and 100 mg/kg cyclophosphamide delivered 25 Gy to critical normal organs. The estimated overall survival (OS) and progression-free survival (PFS) of all treated patients at 2 years was 83% and 68%, respectively. These findings compare favorably with those in a nonrandomized control group of patients who underwent transplantation, external-beam total-body irradiation, and etoposide and cyclophosphamide therapy during the same period (OS of 53% and PFS of 36% at 2 years), even after adjustment for confounding variables in a multivariable analysis. PMID- 11049970 TI - Thalidomide and its analogs overcome drug resistance of human multiple myeloma cells to conventional therapy. AB - Although thalidomide (Thal) was initially used to treat multiple myeloma (MM) because of its known antiangiogenic effects, the mechanism of its anti-MM activity is unclear. These studies demonstrate clinical activity of Thal against MM that is refractory to conventional therapy and delineate mechanisms of anti tumor activity of Thal and its potent analogs (immunomodulatory drugs [IMiDs]). Importantly, these agents act directly, by inducing apoptosis or G1 growth arrest, in MM cell lines and in patient MM cells that are resistant to melphalan, doxorubicin, and dexamethasone (Dex). Moreover, Thal and the IMiDs enhance the anti-MM activity of Dex and, conversely, are inhibited by interleukin 6. As for Dex, apoptotic signaling triggered by Thal and the IMiDs is associated with activation of related adhesion focal tyrosine kinase. These studies establish the framework for the development and testing of Thal and the IMiDs in a new treatment paradigm to target both the tumor cell and the microenvironment, overcome classical drug resistance, and achieve improved outcome in this presently incurable disease. PMID- 11049971 TI - Mechanisms of HIV-associated lymphocyte apoptosis. AB - Infection with the human immunodeficiency virus (HIV) is associated with a progressive decrease in CD4 T-cell number and a consequent impairment in host immune defenses. Analysis of T cells from patients infected with HIV, or of T cells infected in vitro with HIV, demonstrates a significant fraction of both infected and uninfected cells dying by apoptosis. The many mechanisms that contribute to HIV-associated lymphocyte apoptosis include chronic immunologic activation; gp120/160 ligation of the CD4 receptor; enhanced production of cytotoxic ligands or viral proteins by monocytes, macrophages, B cells, and CD8 T cells from HIV-infected patients that kill uninfected CD4 T cells; and direct infection of target cells by HIV, resulting in apoptosis. Although HIV infection results in T-cell apoptosis, under some circumstances HIV infection of resting T cells or macrophages does not result in apoptosis; this may be a critical step in the development of viral reservoirs. Recent therapies for HIV effectively reduce lymphoid and peripheral T-cell apoptosis, reduce viral replication, and enhance cellular immune competence; however, they do not alter viral reservoirs. Further understanding the regulation of apoptosis in HIV disease is required to develop novel immune-based therapies aimed at modifying HIV-induced apoptosis to the benefit of patients infected with HIV. PMID- 11049972 TI - Down-regulation of neutrophil functions by the ELR(+) CXC chemokine platelet basic protein. AB - The platelet-derived neutrophil-activating peptide 2 (NAP-2, 70 amino acids) belongs to the ELR(+) CXC subfamily of chemokines. Similar to other members of this group, such as IL-8, NAP-2 activates chemotaxis and degranulation in neutrophils (polymorphonuclear [PMN]) through chemokine receptors CXCR-1 and CXCR 2. However, platelets do not secrete NAP-2 as an active chemokine but as the C terminal part of several precursors that lack PMN-stimulating capacity. As we have previously shown, PMN themselves may liberate NAP-2 from the precursor connective tissue-activating peptide III (CTAP-III, 85 amino acids) by proteolysis. Instead of inducing cell activation, continuous accumulation of the chemokine in the surroundings of the processing cells results in the down regulation of specific surface-expressed NAP-2 binding sites and in the desensitization of chemokine-induced PMN degranulation. Thus, NAP-2 precursors may be regarded as indirect mediators of functional desensitization in neutrophils. In the current study we investigated the biologic impact of another major NAP-2 precursor, the platelet basic protein (PBP, 94 amino acids). We show that PBP is considerably more potent than CTAP-III to desensitize degranulation and chemotaxis in neutrophils. We present data suggesting that the high desensitizing capacity of PBP is based on its enhanced proteolytic cleavage into NAP-2 by neutrophil-expressed cathepsin G and that it involves efficient down regulation of surface-expressed CXCR-2 while CXCR-1 is hardly affected. Correspondingly, we found PBP and, less potently, CTAP-III to inhibit CXCR-2- but not CXCR-1- dependent chemotaxis of neutrophils toward NAP-2. Altogether our findings demonstrate that the anti-inflammatory capacity of NAP-2 is governed by the species of its precursors. PMID- 11049973 TI - T-lymphocyte production of macrophage inflammatory protein-1alpha is critical to the recruitment of CD8(+) T cells to the liver, lung, and spleen during graft versus-host disease. AB - To investigate the mechanism by which macrophage inflammatory protein-1alpha (MIP 1alpha) affects graft-versus-host disease (GVHD), the expression and function of MIP-1alpha in 2 murine models of GVHD were evaluated. In irradiated class I and class II disparate recipients, the expression of messenger RNA (mRNA) and protein for MIP-1alpha was significantly increased in GVHD target organs after transfer of allogeneic lymphocytes compared to syngeneic lymphocytes. When lymphocytes unable to make MIP-1alpha were transferred, there was a decrease in the production of MIP-1alpha in the liver, lung, and spleen of bm1 (B6.C-H2(bm1)/By) and bm12 (B6.C-H2(bm12)/KhEg) recipients compared to the transfer of wild-type splenocytes. At day 6 there was a 4-fold decrease in the number of transferred CD8(+) T cells in the lung and approximately a 2-fold decrease in the number of CD8(+) T cells in the liver and spleen in bm1 recipients after transfer of MIP 1alpha-deficient (MIP-1alpha(-/-)) splenocytes compared to wild-type (MIP 1alpha(+/+)) splenocytes. These differences persisted for 13 days after splenocyte transfer. In contrast, the number of donor CD4(+) T cells found in the liver and lung was significantly increased after the transfer of MIP-1alpha(-/-) compared to wild-type splenocytes in bm12 recipients from day 6 through day 10. Thus, the transfer of allogeneic T cells was associated with the enhanced expression of MIP-1alpha in both a class I and class II mismatch setting. However, the increased expression only led to enhanced recruitment of CD8(+), but not CD4(+), donor T cells. Production of MIP-1alpha by donor T cells is important in the occurrence of GVHD and functions in a tissue-dependent fashion. PMID- 11049975 TI - Diagnostic value of dominant T-cell clones in peripheral blood in 363 patients presenting consecutively with a clinical suspicion of cutaneous lymphoma. AB - It is now widely accepted that polymerase chain reaction (PCR) analysis of cutaneous T-cell clonality is of diagnostic value in cutaneous T-cell lymphomas (CTCLs) and most helpful in the diagnosis of mycosis fungoides (MF). However, the diagnostic and prognostic value of circulating clonal T cells remains unclear. We studied T-cell clonality in the peripheral blood (PB) and the cutaneous lesion, sampled at the same time, in 363 consecutively seen patients with a clinical suspicion of cutaneous lymphoma. Using a PCR technique providing a specific imprint of T-cell clones (PCRgamma-denaturing gradient gel electrophoresis), we found that detection of identical circulating and cutaneous T-cell clones was associated with the diagnosis of CTCL (P <.001). Detection of circulating tumor cells in patients with MF was infrequent (12.5%), except in those with erythrodermic MF (42%; P =.003). Moreover, among the 46 patients who had identical circulating and cutaneous T-cell clones, 25 (56%) had erythroderma. The finding of a dominant clone in the PB but not in the skin was frequent, regardless of the clinicohistologic classification; it occurred in 30% of patients with CTCL, 41% with non-CTCL malignant infiltrates, and 34% with benign infiltrates. This pattern was significantly more frequent in patients over 60 years of age (P <.002), even in the CTCL group (P <. 01). In conclusion, dominant T-cell clones detected in the PB of patients with MF by using a routine PCR technique are rarely tumoral and are more often related to age. A multicenter prospective study is under way to establish the prognostic value of circulating tumor cells. PMID- 11049974 TI - Long-term follow-up of remission duration, mortality, and second malignancies in hairy cell leukemia patients treated with pentostatin. AB - The nucleoside analogue, pentostatin, has demonstrated high complete response rates and long relapse-free survival times in patients with hairy cell leukemia, a disease that historically had been unresponsive to treatment. Long-term data on duration of overall survival and relapse-free survival and incidence of subsequent malignancies with this agent are lacking. Patients completing the treatment phase of a randomized, intergroup study who received pentostatin as an initial treatment or who crossed over after failure of interferon alpha were followed for survival, relapse, and diagnosis of subsequent malignancies. Two hundred forty-one patients treated with pentostatin as initial therapy (n = 154) or who crossed over after failure of interferon alpha (n = 87) were followed for a median duration of 9.3 years. Estimated 5- and 10-year survival rates (95% confidence interval) for all patients combined were 90% (87%-94%) and 81% (75% 86%), respectively. In the 173 patients with a confirmed complete response to pentostatin treatment, 5- and 10-year relapse-free survival rates were 85% (80% 91%) and 67% (58%-76%), respectively. Survival curves for patients initially treated with pentostatin and those crossed over were similar. Only 2 of 40 deaths were attributed to hairy cell leukemia. The mortality rate and incidence of subsequent malignancies were not higher than expected in the general population. Pentostatin is a highly effective regimen for hairy cell leukemia that produces durable complete responses. Subsequent malignancies do not appear to be increased with pentostatin treatment. PMID- 11049976 TI - Prognostic significance of CD56 expression for ALK-positive and ALK-negative anaplastic large-cell lymphoma of T/null cell phenotype. AB - Anaplastic large cell lymphoma (ALCL) is a distinct entity of non-Hodgkin lymphoma, characterized by a proliferation of pleomorphic large lymphoid cells that express CD30. Recent studies have found that a subset of ALCL aberrantly expresses a chimeric anaplastic lymphoma kinase (ALK) protein as a result of t(2;5)(p23;q35) or variant translocations. ALK-positive ALCLs feature good prognosis, but some of them lead to poor outcomes. Since CD56 is expressed in some ALCLs, its clinical significance was examined in a series of T/null cell type ALCLs. Of 143 patients, 83 (58%) showed ALK-positive staining, and of 140 patients, 25 (18%) expressed CD56. The ALK-positive subgroup was characterized by a younger age of onset (P <.0001), lower serum lactate dehydrogenase level (P =.01), better performance status (P =.03), less frequent extranodal involvement (P =.01), lower international prognostic index (IPI) categories (P =.002), and superior survival (P =.0009) in comparison with the ALK-negative group, suggesting that ALK is a specific marker defining a distinct subtype. CD56(+) cases showed a significantly poor prognosis overall (P =.002) as well as in both ALK-positive and ALK-negative subgroups (P =.02 and P =.04, respectively). Multivariate analysis confirmed that CD56 is independent of other prognostic factors, including IPI. Although CD56(+) cases showed a higher incidence of bone involvement, no other differences in clinicopathologic parameters were found between the CD56(+) and CD56(-) groups. These findings suggest that CD56 is not a marker to identify a distinct subtype of ALCL, but a strong clinical prognostic factor. Effective therapeutic approaches should be explored for high-risk ALCL patients, who can be identified by means of a prognostic model, including CD56. PMID- 11049977 TI - Ex vivo expanded peripheral blood progenitor cells provide rapid neutrophil recovery after high-dose chemotherapy in patients with breast cancer. AB - Ex vivo expanded peripheral blood progenitor cells (PBPCs) have been proposed as a source of hematopoietic support to decrease or eliminate the period of neutropenia after high-dose chemotherapy. CD34 cells were selected from rhG-CSF mobilized PBPCs from patients with breast cancer and were cultured for 10 days in defined media containing 100 ng/mL each of rhSCF, rhG-CSF, and PEG-rhMGDF in 1 L Teflon bags at 20 000 cells/mL. After culture the cells were washed and reinfused on day 0 of transplantation. On day +1, cohort 1 patients (n = 10) also received an unexpanded CD34-selected PBPC product. These patients engrafted neutrophils (absolute neutrophil count, >500/microL) in a median of 6 (range, 5-14) days. Cohort 2 patients (n = 11), who received expanded PBPCs only, engrafted neutrophils in a median of 8 (range, 4-16) days. In comparison, the median time to neutrophil engraftment in a historical control group of patients (n = 100) was 9 days (range, 7-30 days). All surviving patients are now past the 15-month posttransplantation stage with no evidence of late graft failure. The total number of nucleated cells harvested after expansion culture was shown to be the best predictor of time to neutrophil engraftment, with all patients receiving more than 4 x 10(7) cells/kg, engrafting neutrophils by day 8. No significant effect on platelet recovery was observed in any patient. These data demonstrate that PBPCs expanded under the conditions defined can shorten the time to engraftment of neutrophils compared with historical controls and that the rate of engraftment is related to the dose of expanded cells transplanted. PMID- 11049978 TI - Induction of B-cell tolerance by retroviral gene therapy. AB - The primary immunologic barrier to overcome before clinical xenotransplantation can be successful is rejection mediated by preformed natural antibodies in the host, directed toward a single carbohydrate epitope Galalpha1-3Galbeta1-4GlcNAc-R (alphaGal) present on porcine tissue, encoded for by the enzyme glucosyltransferase UDP galactose:beta-D-galactosyl-1, 4-N-acetyl-D-glucosaminide alpha(1-3)galactosyltransferase (EC 2.4.1. 151) or simply alphaGT. Although we have shown previously that a gene therapy approach could be used to prevent production of natural antibodies specific for alphaGal, the ability to induce and maintain tolerance after rigorous antigen challenge would be required if similar approaches are to be used clinically. Here, we demonstrate in alphaGT knockout mice (GT(0) mice), which, like humans, contain in their serum antibodies that bind alphaGal, that the efficient transduction and expression of a retrovirally transduced alphaGT gene in bone marrow-derived cells induces stable long-term tolerance to the alphaGal epitope. GT(0) mice reconstituted with alphaGT transduced bone marrow cells were unable to produce antibodies that bind alphaGal after extensive immunization with pig cells. Furthermore, using ELISPOT assays, we were unable to detect the presence of B cells that produce alphaGal reactive antibodies after immunization, suggesting that such B cells were eliminated from the immunologic repertoire after gene therapy. Interestingly, after tolerance to alphaGal is induced by gene therapy, the antiporcine non-alphaGal humoral response changes from a predominantly IgM to an IgG response. This suggests that once the natural antibody barrier is eliminated by the induction of tolerance, the antipig response changes to a typical T-cell-dependent response involving isotype switching. Thus, gene therapy approaches may be used to overcome immunologic responses leading to xenograft rejection, and similar gene therapy approaches could be used to overcome autoimmunity. PMID- 11049979 TI - Murine hematopoietic stem cell characterization and its regulation in BM transplantation. AB - Using 5-color fluorescence-activated cell sorting, we isolated a subset of murine pluripotent hematopoietic stem cells (PHSC) with the phenotype Lin(-) Sca(+) kit(+) CD38(+) CD34(-) that appears to fulfill the criteria for most primitive PHSC. In the presence of whole bone marrow (BM) competitor cells, these cells produced reconstitution in lethally irradiated primary, secondary, and tertiary murine transplant recipients over the long term. However, these cells alone could not produce reconstitution in lethally irradiated recipients. Rapid proliferation of these cells after BM transplantation required the assistance of another BM cell subset, which has the phenotype Lin(-) Sca(+) kit(+) CD38(-) CD34(+). PMID- 11049980 TI - Imprinting of insulin-like growth factor 2 is modulated during hematopoiesis. AB - The transcription of insulin-like growth factor 2 (IGF-2) is affected by genomic imprinting, a multistep process through which the parental origin of a gene influences its transcription. The maternal copy of IGF-2 is silenced in most human tissues, but in the choroid plexus and the adult liver both alleles of IGF 2 are expressed. This study shows that though in peripheral blood mononuclear cells IGF-2 shows paternal allele-specific expression, in total bone marrow both alleles are transcribed. This modulation of imprinting is not attributable to use of the P1 promoter, because transcription from the P3 promoter occurred from both alleles. These results suggest that transcriptional recognition of the IGF-2 imprint can be modulated during hematopoiesis and may facilitate the development of in vitro model systems to study the transcriptional recognition of a genomic imprint. PMID- 11049981 TI - Generation of murine dendritic cells from flt3-ligand-supplemented bone marrow cultures. AB - Murine dendritic cells (DCs) can be classified into at least 2 subsets, "myeloid related" (CD11b(bright), CD8alpha(-)) and "lymphoid-related" (CD11b(dull), CD8alpha(+)), but the absolute relationship between the 2 remains unclear. Methods of generating DCs from bone marrow (BM) precursors in vitro typically employ granulocyte-macrophage colony-stimulating factor (GM-CSF) as the principal growth factor, and the resultant DCs exhibit a myeloidlike phenotype. Here we describe a flt3-ligand (FL)-dependent BM culture system that generated DCs with more diverse phenotypic characteristics. Murine BM cells cultured at high density in recombinant human FL for 9 days developed into small lymphoid-sized cells, most of which expressed CD11c, CD86, and major histocompatibility complex (MHC) class II. The CD11c(+) population could be divided into 2 populations on the basis of the level of expression of CD11b, which may represent the putative myeloid- and lymphoid-related subsets. The FL in vitro-derived DCs, when treated with interferon-alpha or lipopolysaccharide during the final 24 hours of culture, expressed an activated phenotype that included up-regulation of MHC class II, CD1d, CD8alpha, CD80, CD86, and CD40. The FL-derived DCs also exhibited potent antigen-processing and antigen-presenting capacity. Neutralizing anti-interleukin 6 (IL-6) antibody, but not anti-GM-CSF, significantly reduced the number of DCs generated in vitro with FL, suggesting that IL-6 has a role in the development of DCs from BM precursors. Stem cell factor, which exhibits some of the same bioactivities as FL, was unable to replace FL to promote DC development in vitro. This culture system will facilitate detailed analysis of murine DC development. PMID- 11049983 TI - Characterization of the protein Z-dependent protease inhibitor. AB - Protein Z-dependent protease inhibitor (ZPI) is a 72-kd member of the serpin superfamily of proteinase inhibitors that produces rapid inhibition of factor Xa in the presence of protein Z (PZ), procoagulant phospholipids, and Ca(++) (t(1/2) less than 10 seconds). The rate of factor Xa inhibition by ZPI is reduced more than 1000-fold in the absence of PZ. The factor Xa-ZPI complex is not stable to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but is detectable by alkaline-polyacrylamide gel electrophoresis. The combination of PZ and ZPI dramatically delays the initiation and reduces the ultimate rate of thrombin generation in mixtures containing prothrombin, factor V, phospholipids, and Ca(++). In similar mixtures containing factor Va, however, PZ and ZPI do not inhibit thrombin generation. Thus, the major effect of PZ and ZPI is to dampen the coagulation response prior to the formation of the prothrombinase complex. Besides factor Xa, ZPI also inhibits factor XIa in the absence of PZ, phospholipids, and Ca(++). Heparin (0.2 U/mL) enhances the rate (t(1/2) = 25 seconds vs 50 seconds) and the extent (99% vs 93% at 30 minutes) of factor XIa inhibition by ZPI. During its inhibitory interaction with factor Xa and factor XIa, ZPI is proteolytically cleaved with the release of a 4.2-kd peptide. The N terminal amino acid sequence of this peptide (SMPPVIKVDRPF) establishes Y387 as the P(1) residue at the reactive center of ZPI. ZPI activity is consumed during the in vitro coagulation of plasma through a proteolytic process that involves the actions of factor Xa with PZ and factor XIa. PMID- 11049982 TI - A minimal c-fes cassette directs myeloid-specific expression in transgenic mice. AB - The c-fes proto-oncogene encodes a 92-kd protein tyrosine kinase whose expression is restricted largely to myeloid and endothelial cells in adult mammals. A 13.2 kilobase (kb) human c-fes genomic fragment was previously shown to contain cis acting element(s) sufficient for a locus control function in bone marrow macrophages. Locus control regions (LCRs) confer transgene expression in mice that is integration site independent, copy number dependent, and similar to endogenous murine messenger RNA levels. To identify sequences required for this LCR, c-fes transgenes were analyzed in mice. Myeloid-cell-specific, deoxyribonuclease-I-hypersensitive sites localized to the 3' boundary of exon 1 and intron 3 are required to confer high-level transgene expression comparable to endogenous c-fes, independent of integration site. We define a minimal LCR element as DNA sequences (nucleotides +28 to +2523 relative to the transcription start site) located within intron 1 to intron 3 of the human locus. When this 2.5 kb DNA fragment was linked to a c-fes complementary DNA regulated by its own 446 base-pair promoter, integration-site-independent, copy-number-dependent transcription was observed in myeloid cells in transgenic mice. Furthermore, this 2.5-kb cassette directed expression of a heterologous gene (enhanced green fluorescent protein) exclusively in myeloid cells. The c-fes regulatory unit represents a novel reagent for targeting gene expression to macrophages and neutrophils in transgenic mice. PMID- 11049984 TI - Smooth muscle cell surface tissue factor pathway activation by oxidized low density lipoprotein requires cellular lipid peroxidation. AB - Tissue factor, which is expressed in vascular lesions, increases thrombin production, blood coagulation, and smooth muscle cell proliferation. We demonstrate that oxidized low-density lipoprotein (LDL) induces surface tissue factor pathway activity (ie, activity of the tissue factor:factor VIIa complex) on human and rat smooth muscle cells. Tissue factor messenger RNA (mRNA) was induced by oxidized LDL or native LDL; however, native LDL did not markedly increase tissue factor activity. We hypothesized that oxidized LDL mediated the activation of the tissue factor pathway via an oxidant-dependent mechanism, because antioxidants blocked the enhanced tissue factor pathway activity by oxidized LDL, but not the increased mRNA or protein induction. We separated total lipid extracts of oxidized LDL using high-performance liquid chromatography (HPLC). This yielded 2 major peaks that induced tissue factor activity. Of the known oxysterols contained in the first peak, 7alpha- or 7beta-hydroxy or 7 ketocholesterol had no effect on tissue factor pathway activity; however, 7beta hydroperoxycholesterol increased tissue factor pathway activity without induction of tissue factor mRNA. Tertiary butyl hydroperoxide also increased tissue factor pathway activity, suggesting that lipid hydroperoxides, some of which exist in atherosclerotic lesions, activate the tissue factor pathway. We speculate that thrombin production could be elevated via a mechanism involving peroxidation of cellular lipids, contributing to arterial thrombosis after plaque rupture. Our data suggest a mechanism by which antioxidants may offer a clinical benefit in acute coronary syndrome and restenosis. PMID- 11049985 TI - Platelet release of trimolecular complex components MT1-MMP/TIMP2/MMP2: involvement in MMP2 activation and platelet aggregation. AB - Matrix metalloproteinase 2 (MMP2) has been reported to be secreted by collagen stimulated platelets, and active MMP2 has been shown to play a role in platelet aggregation. It has been demonstrated that MMP2 activation is dependent on the complex (membrane type 1 [MT1]-MMP/tissue inhibitor of MMP2 [TIMP2]) receptor and MMP2. We have investigated human platelets as a possible source of MT1-MMP, and we have studied its role in MMP2 activation and in platelet aggregation. Gelatin zymograms showed the existence of MMP2 at proforms (68 kd) and activated-enzyme forms (62-59 kd) in supernatants of resting and activated platelets, respectively. No gelatinolytic activity was associated with the platelet pellet after aggregation, suggesting a total release of MMP2 during cell activation. By Western blot analysis in nonreduced conditions, MT1-MMP was found on resting platelet membranes in 2 forms-the inactive 45-kd form and an apparent 89-kd form, which totally disappeared under reduced conditions. After platelet degranulation, only the 45-kd form was detected. Reverse transcription-polymerase chain reaction experiments showed the expression in platelets of messenger RNA encoding for MMP2, MT1-MMP, and TIMP2. Flow cytometry analysis showed that MT1-MMP, MMP2, and TIMP2 expressions were enhanced at the activated platelet surface. MMP inhibitors, recombinant TIMP2, and synthetic BB94 inhibited collagen-induced platelet aggregation in a concentration-dependent manner, indicating the role of activated MT1-MMP in the modulation of platelet function. In conclusion, our results demonstrate the expression of the trimolecular complex components (MT1 MMP/TIMP2/MMP2) by blood platelets as well as the ability of MMP inhibitors to modulate the aggregating response. PMID- 11049986 TI - Dimerization of P-selectin in platelets and endothelial cells. AB - P-selectin is a leukocyte adhesion receptor stored in platelets and endothelial cells and is translocated to the surface upon cell activation. Purified P selectin is oligomeric and has increased avidity for its ligand relative to the monomeric form, but whether P-selectin self-associates in the membrane of intact cells is not known. A chemical cross-linking approach was used to show that P selectin is present as noncovalent dimers in resting platelets, human umbilical vein endothelial cells, and heterologous RIN5F cells expressing P-selectin. The results of 2-dimensional isoelectric focusing are consistent in showing P selectin dimers as homodimers, but they are composed of a more basic subset of P selectin than the monomers. This suggests that the dimers are a biochemically distinct subset of P-selectin. P-selectin dimers form in the endoplasmic reticulum and Golgi compartments of human umbilical vein endothelial cells only after synthesis of the mature P-selectin subunit, and are not preferentially stored in Weibel-Palade bodies as compared with the monomeric form. Platelet activation with thrombin receptor-activating peptide leads to the presence of P selectin monomers and homodimers on the cell surface as well as P-selectin heterodimers, which are composed of P-selectin and an unidentified protein of approximately 81 kd molecular weight. In summary, these studies demonstrate that P-selectin is homodimeric in situ and that platelet activation leads to the formation of an additional activation-specific heterodimeric species. In addition, the homodimer has unique biochemical characteristics compared with the monomeric form, and dimerization occurs in the endoplasmic reticulum and Golgi compartments of endothelial cells. PMID- 11049987 TI - Role of SCL/Tal-1, GATA, and ets transcription factor binding sites for the regulation of flk-1 expression during murine vascular development. AB - The receptor tyrosine kinase Flk-1 is essential for embryonic blood vessel development and for tumor angiogenesis. To identify upstream transcriptional regulators of Flk-1, the gene regulatory elements that mediate endothelium specific expression in mouse embryos were characterized. By mutational analysis, binding sites for SCL/Tal-1, GATA, and Ets transcription factors located in the Flk-1 enhancer were identified as critical elements for the endothelium-specific Flk-1 gene expression in transgenic mice. c-Ets1, a transcription factor that is coexpressed with Flk-1 during embryonic development and tumor angiogenesis, activated the Flk-1 promoter via 2 binding sites. One of these sites was required for Flk-1 promoter function in the embryonic vasculature. These results provide the first evidence that SCL/Tal-1, GATA, and Ets transcription factors act upstream of Flk-1 in a combinatorial fashion to determine embryonic blood vessel formation and are key regulators not only of the hematopoietic program, but also of vascular development. PMID- 11049988 TI - The human antimicrobial and chemotactic peptides LL-37 and alpha-defensins are expressed by specific lymphocyte and monocyte populations. AB - We identified antibacterial components in human T and natural killer (NK) cells by using freshly isolated lymphocytes enriched for T and NK cells as starting material. After growing these lymphocytes for 5 days in the presence of interleukin (IL)-2, we isolated and characterized several antibacterial peptides/proteins from the supernatant-alpha-defensins (HNP 1-3), LL-37, lysozyme, and a fragment of histone H2B-although other active components were also present. We then used reverse transcriptase-polymerase chain reaction to search for expression of the gene coding for LL-37 in several B-cell lines, gammadelta T-cell lines, NK clones, and one monocytic cell line, with positive results, but found no expression in several alphabeta T-cell lines. The alpha defensins (HNP 1-3) were also found to be expressed in several of these cell lines. To confirm the presence of these antibacterial peptides in lymphocytes, we localized them to NK, gammadelta T cells, B cells, and monocytes/macrophages by using double-staining immunohistochemical analysis of freshly isolated lymphocytes. We also found that primary cultures of lymphocytes transcribe and secrete LL-37 and that these processes are affected by IL-6 and interferon-gamma. In addition, we demonstrated that LL-37 has chemotactic activity for polymorphonuclear leukocytes and CD4 T lymphocytes, whereas others have shown chemotactic activity for human alpha-defensins (HNP 1-2). These findings suggest that microbicidal peptides are effector molecules of lymphocytes and that antibacterial activity previously shown to be derived from T and NK cells may be partly mediated by the antibacterial peptides LL-37 and HNP 1-3. PMID- 11049989 TI - Impaired function of circulating HIV-specific CD8(+) T cells in chronic human immunodeficiency virus infection. AB - The functional status of circulating human immunodeficiency (HIV)-specific CD8 T cells in chronically infected subjects was evaluated. By flow cytometry, only 5 of 7 subjects had detectable CD8 T cells that produced IFN-gamma after stimulation with HIV-infected primary CD4 T cells. In 2 subjects, the frequency of IFN-gamma-producing cells increased 4-fold when IL-2 was added to the culture medium; in another subject, IFN-gamma-producing cells could be detected only after IL-2 was added. IFN-gamma-producing cells ranged from 0.4% to 3% of CD8 T cells. Major histocompatibility complex-peptide tetramer staining, which identifies antigen-specific T cells irrespective of function, was used to evaluate the proportion of HIV-specific CD8 T cells that may be nonfunctional in vivo. CD8 T cells binding to tetramers complexed to HIV gag epitope SLYNTVATL and reverse transcriptase epitope YTAFTIPSI were identified in 9 of 15 and 5 of 12 HLA-A2-expressing seropositive subjects at frequencies of 0.1% to 1.1% and 0.1 to 0.7%, respectively. Freshly isolated tetramer-positive cells expressed a mixed pattern of memory and effector markers. On average, IFN-gamma was produced by less than 25% of tetramer-positive CD8 T cells after stimulation with the relevant gag or reverse transcriptase peptide. In all subjects tested, freshly isolated CD8 T cells were not cytolytic against peptide-pulsed B lymphoblastoid cell line or primary HIV-infected CD4 T-cell targets. Exposure to IL-2 enhanced the cytotoxicity of CD8 T cells against primary HIV-infected CD4 targets in 2 of 2 subjects tested. These results suggest that a significant proportion of HIV specific CD8 T cells may be functionally compromised in vivo and that some function can be restored by exposure to IL-2. PMID- 11049990 TI - Induction of cytotoxic T-lymphocyte responses in vivo after vaccinations with peptide-pulsed dendritic cells. AB - Vaccination of patients with cancer using dendritic cells (DCs) was shown to be effective for B-cell lymphoma and malignant melanoma. Here we provide evidence that patients with advanced breast and ovarian cancer can be efficiently vaccinated with autologous DCs pulsed with HER-2/neu- or MUC1-derived peptides. Ten patients were included in this pilot study. The DC vaccinations were well tolerated with no side effects. In 5 of 10 patients, peptide-specific cytotoxic T lymphocytes (CTLs) could be detected in the peripheral blood using both intracellular IFN-gamma staining and (51)Cr-release assays. The major CTL response in vivo was induced with the HER-2/neu-derived E75 and the MUC1-derived M1.2 peptide, which lasted for more than 6 months, suggesting that these peptides might be immunodominant. In addition, in one patient vaccinated with the MUC1 derived peptides, CEA- and MAGE-3 peptide-specific T-cell responses were detected after several vaccinations. In a second patient immunized with the HER-2/neu peptides, MUC1-specific T lymphocytes were induced after 7 immunizations, suggesting that antigen spreading in vivo might occur after successful immunization with a single tumor antigen. Our results show that vaccination of DCs pulsed with a single tumor antigen may induce immunologic responses in patients with breast and ovarian cancer. This study may be relevant to the design of future clinical trials of other peptide-based vaccines. PMID- 11049991 TI - Interferon gamma and interleukin 6 modulate the susceptibility of macrophages to human immunodeficiency virus type 1 infection. AB - The effect of interferon gamma (IFN-gamma) and interleukin 6 (IL-6) on infection of macrophages with human immunodeficiency virus type 1 (HIV-1) was investigated. By using a polymerase chain reaction-based viral entry assay and viral infectivity assay, it was demonstrated that IL-6 and IFN-gamma augmented susceptibility of monocyte-derived macrophages (MDMs) to infection with T-cell tropic CXCR4-utilizing (X4) HIV-1 strains. Consistent with this finding, IFN gamma and IL-6 augmented fusion of MDMs with T-tropic envelope-expressing cells. The enhanced fusion of cytokine-treated MDMs with T-tropic envelopes was inhibited by the CXCR4 ligand, SDF-1, and by T22 peptide. IFN-gamma and IL-6 did not affect expression of surface CXCR4 or SDF-1-induced Ca(++) flux in MDMs. In contrast to the effect of IFN-gamma on the infection of MDMs with X4 strains, IFN gamma inhibited viral entry and productive infection of MDMs with macrophage tropic (M-tropic) HIV-1. Consistent with this finding, IFN-gamma induced a decrease in fusion with M-tropic envelopes that correlated with a modest reduction in surface CCR5 and CD4 on MDMs. It was further demonstrated that macrophage inflammatory protein (MIP)-1alpha and MIP-beta secreted by cytokine treated MDMs augmented their fusion with T-tropic-expressing cells and inhibited their fusion with M-tropic envelope-expressing cells. These data indicate that proinflammatory cytokines, which are produced during opportunistic infections or sexually transmitted diseases, may predispose macrophages to infection with X4 strains that, in turn, could accelerate disease progression. PMID- 11049992 TI - Correlation of mutations of the SH2D1A gene and epstein-barr virus infection with clinical phenotype and outcome in X-linked lymphoproliferative disease. AB - The purposes of this study were to determine the frequency of mutations in SH2D1A in X-linked lymphoproliferative disease (XLP) and the role of SH2D1A mutations and Epstein-Barr virus (EBV) infection in determining the phenotype and outcome of patients with XLP. Analysis of 35 families from the XLP Registry revealed 28 different mutations in 34 families-large genomic deletions (n = 3), small intragenic deletions (n = 10), splice-site (n = 3), nonsense (n = 3), and missense (n = 9) mutations. No mutations were found in 25 males, so-called sporadic XLP (males with an XLP phenotype after EBV infection but no family history of XLP) or in 9 patients with chronic active EBV syndrome. Of 304 symptomatic males in the XLP Registry, 38 had no evidence of EBV infection at first clinical manifestation. When fulminant infectious mononucleosis (FIM) was excluded, there was no statistical difference in the frequency of EBV infectivity in the other XLP phenotypes. Furthermore, there was no difference at age of first clinical manifestation between EBV(+) and EBV(-) males or in survival when patients with FIM were excluded. In conclusion, it was found that mutations in the SH2D1A gene are responsible for XLP but that there is no correlation between genotype and phenotype or outcome. It was also found that though EBV infection often results in FIM, it is unnecessary for the expression of other manifestations of XLP, and it correlates poorly with outcome. These results suggest that unidentified factors, either environmental or genetic (eg, modifier genes), contribute to the pathogenesis of XLP. PMID- 11049993 TI - UTY gene codes for an HLA-B60-restricted human male-specific minor histocompatibility antigen involved in stem cell graft rejection: characterization of the critical polymorphic amino acid residues for T-cell recognition. AB - Rejection of a graft after human leukocyte antigen (HLA)-identical stem cell transplantation (SCT) can be caused by recipient's immunocompetent T lymphocytes recognizing minor histocompatibility antigens on donor stem cells. During rejection of a male stem cell graft by a female recipient, 2 male (H-Y)-specific cytotoxic T lymphocyte (CTL) clones were isolated from peripheral blood. One CTL clone recognized an HLA-A2-restricted H-Y antigen, encoded by the SMCY gene. Another CTL clone recognized an HLA-B60-restricted H-Y antigen. In this study UTY was identified as the gene coding for the HLA-B60-restricted H-Y antigen. The UTY derived H-Y antigen was characterized as a 10-amino acid residue peptide, RESEEESVSL. Although the epitope differed by 3 amino acids from its X-homologue, UTX, only 2 polymorphisms were essential for recognition by the CTL clone HLA-B60 HY. These results illustrate that CTLs against several H-Y antigens derived from different proteins can contribute simultaneously to graft rejection after HLA identical, sex-mismatched SCT. Moreover, RESEEESVSL-specific T cells could be isolated from a female HLA-B60+ patient with myelodysplastic syndrome who has been treated with multiple blood transfusions, but not from control healthy HLA B60+ female donors. This may indicate that RESEEESVSL-reactive T cells are more common in sensitized patients. PMID- 11049994 TI - Molecular single-cell analysis of the clonal relationship of small Epstein-Barr virus-infected cells and Epstein-Barr virus-harboring Hodgkin and Reed/Sternberg cells in Hodgkin disease. AB - Epstein-Barr virus (EBV) can be detected in the tumor cells of approximately 40% of cases of classical Hodgkin disease (cHD). Clonality studies suggest that infection of the neoplastic Hodgkin and Reed/Sternberg (HRS) cells occurs before tumor clone expansion. In EBV-positive cases, variable numbers of EBER-positive small B cells are sometimes also observed that immunohistologically differ from the neoplastic cells by lack of CD30 and latent membrane protein 1 expression. To analyze the clonal relationship between these EBV(+) cells and the HRS cells, single EBV-infected CD30(-) B cells, as well as HRS cells from 3 cases of EBV positive cHD were micromanipulated, their immunoglobulin gene rearrangements amplified and then compared with each other. In 2 cases, all small EBV-infected cells were clonally unrelated to the HRS cells. In a third case, 2 of 29 small CD30(-) cells were found to carry HRS cell-specific rearrangements. Thus, small CD30(-) EBV-infected B cells in cHD belong to the HRS tumor clone rarely, if at all. In all cases, small clones unrelated to the HRS cell clones were identified among the small EBV(+) CD30(-) cells. The vast majority of small EBV(+) CD30(-) B cells was found to carry somatically mutated V region genes, indicating that in lymph nodes of patients with HD, like in the peripheral blood of healthy individuals, EBV persists in memory B cells. PMID- 11049995 TI - High levels of soluble syndecan-1 in myeloma-derived bone marrow: modulation of hepatocyte growth factor activity. AB - Syndecan-1 is a heparan sulfate proteoglycan expressed on the surface of, and actively shed by, myeloma cells. Hepatocyte growth factor (HGF) is a cytokine produced by myeloma cells. Previous studies have demonstrated elevated levels of syndecan-1 and HGF in the serum of patients with myeloma, both of negative prognostic value for the disease. Here we show that the median concentrations of syndecan-1 (900 ng/mL) and HGF (6 ng/mL) in the marrow compartment of patients with myeloma are highly elevated compared with healthy controls and controls with other diseases. We show that syndecan-1 isolated from the marrow of patients with myeloma seems to exist in an intact form, with glucosaminoglycan chains. Because HGF is a heparan-sulfate binding cytokine, we examined whether it interacted with soluble syndecan-1. In supernatants from myeloma cells in culture as well as in pleural effusions from patients with myeloma, HGF existed in a complex with soluble syndecan-1. Washing myeloma cells with purified soluble syndecan-1 could effectively displace HGF from the cell surface, suggesting that soluble syndecan 1 can act as a carrier for HGF in vivo. Finally, using a sensitive HGF bioassay (interleukin-11 production from the osteosarcoma cell line Saos-2) and intact syndecan-1 isolated from the U-266 myeloma cell line, we found that the presence of high concentrations of syndecan-1 (more than 3 microg/mL) inhibited the HGF effect, whereas lower concentrations potentiated it. HGF is only one of several heparin-binding cytokines associated with myeloma. These data indicate that soluble syndecan-1 may participate in the pathology of myeloma by modulating cytokine activity within the bone marrow. PMID- 11049996 TI - Elevated soluble MUC1 levels and decreased anti-MUC1 antibody levels in patients with multiple myeloma. AB - Soluble MUC1 (sMUC1) levels are elevated in many MUC1(+) cancers. We and others have shown that MUC1 is expressed on multiple myeloma (MM) plasma cells and B cells. In this study, we measured sMUC1 levels in bone marrow (BM) plasma from 71 MM patients and 21 healthy donors (HDs), and in peripheral blood (PB) plasma from 42 MM patients and 13 HDs using an immunoassay that detects the CA27.29 epitope of MUC1. sMUC1 levels were found to be significantly greater (mean 31.76 U/mL, range 5.69 to 142.48 U/mL) in MM patient BM plasma versus HD BM plasma (mean 9.68 U/mL, range 0.65 to 39.83 U/mL) (P <. 001). Importantly, BM plasma sMUC1 levels were related to tumor burden because sMUC1 levels were significantly higher for MM patients with active disease (34.62 U/mL, range 5.69 to 142.48 U/mL) versus MM patients with minimal residual disease (16.16 U/mL, range 5.7 to 56.68 U/mL) (P =.0026). sMUC1 levels were also elevated in the PB plasma of MM patients (32.79 U/mL, range 4.15 to 148.84 U/mL) versus HDs (18.47 U/mL, range 8.84 to 42.49) (P =.0052). Lastly, circulating immunglobulin M (IgM) and IgG antibodies to MUC1 were measured in 114 MM patients and 31 HDs, because natural antibodies to MUC1 have been detected in patients with other MUC1-bearing malignancies. These studies demonstrated lower levels of circulating IgM (P <.001) and IgG (P =.078) antibodies to MUC1 in MM patients compared with HDs. Our data therefore show that in MM patients, sMUC1 levels are elevated and correlate with disease burden, whereas anti-MUC1 antibody levels are decreased. PMID- 11049997 TI - Mutations of the AML1 gene in myelodysplastic syndrome and their functional implications in leukemogenesis. AB - The AML1 gene encodes a DNA-binding protein that contains the runt domain and is the most frequent target of translocations associated with human leukemias. Here, point mutations of the AML1 gene, V105ter (single-letter amino acid code) and R139G, (single-letter amino acid codes) were identified in 2 cases of myelodysplastic syndrome (MDS) by means of the reverse transcriptase-polymerase chain reaction single-strand conformation polymorphism method. Both mutations are present in the region encoding the runt domain of AML1 and cause loss of the DNA binding ability of the resultant products. Of these mutants, V105ter has also lost the ability to heterodimerize with polyomavirus enhancer binding protein 2/core binding factor beta (PEBP2beta/CBFbeta). On the other hand, the R139G mutant acts as a dominant negative inhibitor by competing with wild-type AML1 for interaction with PEBP2beta/CBFbeta. This study is the first report that describes mutations of AML1 in patients with MDS and the mechanism whereby the mutant acts as a dominant negative inhibitor of wild-type AML1. PMID- 11049998 TI - Involvement of CD44-hyaluronan interaction in malignant cell homing and fibronectin synthesis in hairy cell leukemia. AB - The tissue homing of malignant hematic cells has both diagnostic and pathogenetic importance. Although such homing is incompletely understood, it generally involves cell adhesion and migration mediated by a number of adhesion receptors and cytokines. In this article, the potential importance of hyaluronan (HA) is examined for the tissue homing of hairy cells (HCs) in hairy cell leukemia (HCL). It is shown that HCs readily adhere to, and spontaneously move on, HA-coated surfaces using CD44. This indicates that activated CD44 and spontaneous movement on HA form part of the intrinsically activated phenotype of HCs. Interleukin-8 (IL-8) inhibited HC movement on HA, and this cell arrest was accompanied by increased actin polymerization and a more pronounced association of CD44 with the cytoskeleton. All of these findings are in sharp contrast to our previous observations with chronic lymphocytic leukemia cells, which are nonmotile on HA, but in response to IL-8 become polarized and motile using the receptor for HA mediated motility rather than their apparently inactive CD44. Immunohistochemical examination of HCL tissues showed the ubiquitous presence of IL-8 and the prominence of HA in bone marrow stroma and hepatic portal tracts. This suggests that CD44-HA interactions are important in HC homing to these sites, but not to splenic red pulp or hepatic sinusoids, where HA is largely absent. Moreover, engagement of CD44 on HCs stimulates fibronectin synthesis, an observation that is likely to be relevant to the restriction of fibrosis in the disease to HC infiltrated areas containing HA. PMID- 11049999 TI - Chronic lymphocytic leukemia B cells inhibit spontaneous Ig production by autologous bone marrow cells: role of CD95-CD95L interaction. AB - A variable degree of humoral immunodeficiency is a common feature in patients with B-cell chronic lymphocytic leukemia (B-CLL). The aim of this study was to explore the possibility that B-CLL cells play a direct role in this phenomenon. To this end, patients' bone marrow (BM) immunoglobulin (Ig)-secreting cells were cocultured with autologous purified B-CLL cells. The results show that tumoral cells inhibited the spontaneous IgG secretion by BM plasma cells, and this effect increased after PMA-induction of B-CLL cells. This inhibitory process was proportional to the number of B-CLL cells added and depended on cellular contact. Adhesion molecules did not appear to be involved in the cellular interaction, because the inclusion of blocking antibody to a variety of these proteins did not reverse the inhibitory phenomenon. However, the addition of monoclonal antibody that blocked the function of either CD95 or CD95L clearly reversed B-CLL cell inhibition on autologous BM plasma cells. These latter cells were shown to express CD95, and B-CLL cells contained detectable quantities of CD95L at the level of messenger RNA and protein. Annexin V-binding experiments revealed increased apoptosis of BM Ig-secreting cells when cocultured with autologous B CLL cells. Finally, this inhibitory phenomenon might be operative in vivo because (a) there was a good correlation between the intensity of the inhibitory effect in vitro and the serum IgG level exhibited by every patient and (b) B-CLL cells also inhibited in vivo antigen-induced IgG-tetanus toxoid-secreting cells obtained from normal immunized subjects. Collectively, these data suggest that B CLL cells inhibit autologous CD95-bearing Ig-secreting cells by the interaction with CD95L present on B-CLL cells and, hence, contribute to the state of humoral immunodeficiency that occurs in these patients. PMID- 11050000 TI - Heterogeneity in therapeutic response of genetically altered myeloma cell lines to interleukin 6, dexamethasone, doxorubicin, and melphalan. AB - Because there is no known genetic abnormality common to all patients with myeloma, it is important to understand how genetic heterogeneity may lead to differences in signal transduction, cell cycle, and response to therapy. Model cell lines have been used to study the effect that mutations in p53 and ras can have on growth properties and responses of myeloma cells. The U266 cell line has a single mutant p53 allele. Stable expression of wild-type (wt) p53 in U266 cells results in a significant suppression of interleukin (IL)-6 gene expression and in the concomitant suppression of cell growth that could be restored by the addition of exogenous IL-6. Expression of wt p53 also leads to cell cycle arrest and protection from doxorubicin (Dox)- and melphalan (Mel)-induced apoptosis. The addition of IL-6 resulted in cell cycle progression and blocked p53-mediated protection from apoptosis. ANBL6 is an IL-6-dependent cell line that is sensitive to dexamethasone (Dex), Dox, and Mel. IL-6 is able to protect ANBL6 cells from Dex- and Mel- but not Dox-induced apoptosis. To study the effect of an activating mutation in ras, the ANBL6 cell line transfected with either a constitutively activated N- or K-ras gene was used. Both N-ras12 and K-ras12 genes were able to protect ANBL6 cells from apoptosis induced by Dex, Dox, and Mel. These data show that changes in ras or p53 can alter the myeloma cell response to IL-6 and demonstrate that the genetic background can alter therapeutic responses. PMID- 11050001 TI - In vitro and in vivo production of vascular endothelial growth factor by chronic lymphocytic leukemia cells. AB - Expansion of primary solid tumors and their malignant dissemination are angiogenesis-dependent. Vascular endothelial growth factor (VEGF) is the key factor playing a pivotal role in solid tumor-induced angiogenesis. Recent studies indicate that angiogenesis may also be involved in the pathogenesis of certain hemic malignancies, including B-cell chronic lymphocytic leukemia (B-CLL). Mechanisms underlying angiogenesis in B-CLL and the role of VEGF in this process are incompletely understood. In this study, it was examined whether angiogenically functional VEGF is produced by B-CLL cells. Immunohistochemical staining with antibodies against VEGF and CD34, an endothelial cell marker, demonstrated the presence of VEGF protein and abundant blood vessels in infiltrated lymphoreticular tissues. Low levels of VEGF were detected by ELISA in the culture media of unstimulated cells; this was enhanced up to 7-fold by hypoxic stimulation. SDS-PAGE and Western blot analysis of the concentrated culture media showed 2 isoforms of VEGF protein with molecular weights of 28 and 42 kd, respectively. RNA hybridization showed that these cells expressed VEGF mRNA. Reverse transcription-polymerase chain reaction, combined with nucleotide sequence analysis, revealed that the predominantly expressed isoforms were VEGF121 and VEGF165. Moreover, (3)H-thymidine incorporation and an in vivo angiogenic assay demonstrated that the VEGF produced by CLL cells can induce angiogenesis by stimulating endothelial cell proliferation. In conclusion, this study shows that B-CLL cells produce VEGF and demonstrates the angiogenic effects of this growth factor, which may be relevant for the tissue phase of the disease. PMID- 11050002 TI - Kaposi sarcoma is a therapeutic target for vitamin D(3) receptor agonist. AB - Kaposi sarcoma (KS) is responsive to a number of different steroid hormones, such as glucocorticoids and retinoids. An active metabolite of vitamin D, 1alpha,25 dihydroxyvitamin D(3), was used to study the effect of this steroid hormone in KS. Steroid hormones exert their effect through their cognate nuclear receptors, which for vitamin D metabolites is the vitamin D receptor (VDR). It was first shown that KS cell lines and primary tumor tissue express high levels of VDR, whereas endothelial cells had minimal expression and fibroblasts had no expression. Second, KS cell growth was inhibited by VDR agonist 1alpha,25 dihydroxyvitamin D(3) with a 50% inhibitory concentration of 5 x 10 -8 mol/L, whereas endothelial cells and fibroblast cells showed no response. Studies on the mechanism of KS tumor growth inhibition by 1alpha,25 dihydroxyvitamin D(3) showed that production of autocrine growth factors interleukin (IL)-6 and IL-8 was reduced in a dose-dependent manner, whereas no effect was observed on vascular endothelial growth factor and basic fibroblast growth factor. Transcription initiated at the IL-6 promoter was repressed by VDR agonist. The DNA sequences required to mediate this repression were localized to nucleotides -225/-110 in the 5'-flanking region. The antitumor activity of VDR agonists was also confirmed in KS tumor xenograft and after topical application in patients with KS. 1alpha,25 Dihydroxyvitamin D(3) and its analogs may thus be candidates for clinical development in KS. PMID- 11050003 TI - Efficacy of STI571, an abl tyrosine kinase inhibitor, in conjunction with other antileukemic agents against bcr-abl-positive cells. AB - Chronic myelogenous leukemia (CML), a malignancy of a hematopoietic stem cell, is caused by the Bcr-Abl tyrosine kinase. STI571(formerly CGP 57148B), an Abl tyrosine kinase inhibitor, has specific in vitro antileukemic activity against Bcr-Abl-positive cells and is currently in Phase II clinical trials. As it is likely that resistance to a single agent would be observed, combinations of STI571 with other antileukemic agents have been evaluated for activity against Bcr-Abl-positive cell lines and in colony-forming assays in vitro. The specific antileukemic agents tested included several agents currently used for the treatment of CML: interferon-alpha (IFN), hydroxyurea (HU), daunorubicin (DNR), and cytosine arabinoside (Ara-C). In proliferation assays that use Bcr-Abl expressing cells lines, the combination of STI571 with IFN, DNR, and Ara-C showed additive or synergistic effects, whereas the combination of STI571 and HU demonstrated antagonistic effects. However, in colony-forming assays that use CML patient samples, all combinations showed increased antiproliferative effects as compared with STI571 alone. These data indicate that combinations of STI571 with IFN, DNR, or Ara-C may be more useful than STI571 alone in the treatment of CML and suggest consideration of clinical trials of these combinations. PMID- 11050004 TI - Altered ligand binding and transcriptional regulation by mutations in the PML/RARalpha ligand-binding domain arising in retinoic acid-resistant patients with acute promyelocytic leukemia. AB - Acute promyelocytic leukemia (APL) is characterized by a specific translocation, t(15;17), that fuses the promyelocytic leukemia (PML) gene with the RA receptor RARalpha. Pharmacologic doses of retinoic acid (RA) induce differentiation in human APL cells and complete clinical remissions. Unfortunately, APL cells develop resistance to RA in vitro and in vivo. Recently, mutations in PML/RARalpha have been described in APL cells from patients clinically resistant to RA therapy. The mutations cluster in 2 regions that are involved in forming the binding pocket for RA. These mutant PML/RARalpha proteins have been expressed in vitro, which shows that they cause a diversity of alterations in binding to ligand and to nuclear coregulators of transcription, leading to varying degrees of inhibition of retinoid-induced transcription. This contrasts with the nearly complete dominant negative activity of mutations in PML/RARalpha previously characterized in cell lines developing RA resistance in vitro. Current data from this study provide additional insight into the molecular mechanisms of resistance to RA and suggest that alterations in the ability of mutants to interact with coregulators can be determinant in the molecular mechanism of resistance to RA. In particular, ligand-induced binding to the coactivator ACTR correlated better with transcriptional activation of RA response elements than the ligand-induced release of the corepressor SMRT. The diversity of effects that are seen in patient-derived mutations may help explain the partial success to date of attempts to overcome this mechanism of resistance in patients by the clinical use of histone deacetylase inhibitors. PMID- 11050005 TI - A novel gene, MEL1, mapped to 1p36.3 is highly homologous to the MDS1/EVI1 gene and is transcriptionally activated in t(1;3)(p36;q21)-positive leukemia cells. AB - The reciprocal translocation t(1;3)(p36;q21) occurs in a subset of myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML), which is frequently characterized by trilineage dysplasia, in particular dysmegakaryocytopoiesis, and poor prognosis. Previously, the breakpoint cluster region (BCR) at 3q21 was identified within a 60-kilobase (kb) region centromeric to the BCR of 3q21q26 syndrome and that at 1p36.3 within a 90-kb region. In this study, genes were searched near the breakpoints at 1p36.3, and a novel gene was isolated that encoded a zinc finger protein with a PR domain, which is highly homologous to the MDS1/EVI1 gene. The novel gene, designated as MEL1 (MDS1/EVI1 like gene 1), with 1257 amino acid residues is 64% similar in nucleotide and 63% similar in amino acid sequences to MDS1/EVI1 with the same domain structure. The MEL1 gene is expressed in leukemia cells with t(1;3) but not in other cell lines or bone marrow, spleen, and fetal liver, suggesting that MEL1 is specifically in the t(1;3)(p36;q21)-positive MDS/AML. On the basis of the positional relationship between the EVI1 and MEL1 genes in each translocation, it was suggested that both genes are transcriptionally activated by the translocation of the 3q21 region with the Ribophorin I gene. Because of the transcriptional activation of the EVI1 family genes in both t(1;3)(p36;q21)-positive MDS/AML and 3q21q26 syndrome, it is suggested that they share a common molecular mechanism for the leukemogenic transformation of the cells. PMID- 11050006 TI - Identification and characterization of CKLiK, a novel granulocyte Ca(++)/calmodulin-dependent kinase. AB - Human granulocytes are characterized by a variety of specific effector functions involved in host defense. Several widely expressed protein kinases have been implicated in the regulation of these effector functions. A polymerase chain reaction-based strategy was used to identify novel granulocyte-specific kinases. A novel protein kinase complementary DNA with an open reading frame of 357 amino acids was identified with homology to calcium-calmodulin-dependent kinase I (CaMKI). This has been termed CaMKI-like kinase (CKLiK). Analysis of CKLiK messenger RNA (mRNA) expression in hematopoietic cells demonstrated an almost exclusive expression in human polymorphonuclear leukocytes (PMN). Up-regulation of CKLiK mRNA occurs during neutrophilic differentiation of CD34(+) stem cells. CKLiK kinase activity was dependent on Ca(++) and calmodulin as analyzed by in vitro phosphorylation of cyclic adenosine monophosphate responsive element modulator (CREM). Furthermore, CKLiK- transfected cells treated with ionomycin demonstrated an induction of CRE- binding protein (CREB) transcriptional activity compared to control cells. Additionally, CaMK-kinasealpha enhanced CKLiK activity. In vivo activation of CKLiK was shown by addition of interleukin (IL)-8 to a myeloid cell line stably expressing CKLiK. Furthermore inducible activation of CKLiK was sufficient to induce extracellular signal-related kinase (ERK) mitogen-activated protein (MAP) kinase activity. These data identify a novel Ca(++)/calmodulin-dependent PMN- specific kinase that may play a role in Ca(++) mediated regulation of human granulocyte functions. PMID- 11050007 TI - The fanconi anemia proteins FANCA and FANCG stabilize each other and promote the nuclear accumulation of the Fanconi anemia complex. AB - Fanconi anemia (FA) is an autosomal recessive cancer susceptibility syndrome with 8 complementation groups. Four of the FA genes have been cloned, and at least 3 of the encoded proteins, FANCA, FANCC, and FANCG/XRCC9, interact in a multisubunit protein complex. The FANCG protein binds directly to the amino terminal nuclear localization sequence (NLS) of FANCA, suggesting that FANCG plays a role in regulating FANCA nuclear accumulation. In the current study the functional consequences of FANCG/FANCA binding were examined. Correction of an FA G cell line with the FANCG complementary DNA (cDNA) resulted in FANCA/FANCG binding, prolongation of the cellular half-life of FANCA, and an increase in the nuclear accumulation of the FA protein complex. Similar results were obtained upon correction of an FA-A cell line, with a reciprocal increase in the half-life of FANCG. Patient-derived mutant forms of FANCA, containing an intact NLS sequence but point mutations in the carboxy-terminal leucine zipper region, bound FANCG in the cytoplasm. The mutant forms failed to translocate to the nucleus of transduced cells, thereby suggesting a model of coordinated binding and nuclear translocation. These results demonstrate that the FANCA/FANCG interaction is required to maintain the cellular levels of both proteins. Moreover, at least one function of FANCG and FANCA is to regulate the nuclear accumulation of the FA protein complex. Failure to accumulate the nuclear FA protein complex results in the characteristic spectrum of clinical and cellular abnormalities observed in FA. PMID- 11050008 TI - Nonopsonic monocyte/macrophage phagocytosis of Plasmodium falciparum-parasitized erythrocytes: a role for CD36 in malarial clearance. AB - Plasmodium falciparum is the most lethal form of malaria and is increasing both in incidence and in its resistance to antimalarial agents. An improved understanding of the mechanisms of malarial clearance may facilitate the development of new therapeutic interventions. We postulated that the scavenger receptor CD36, an important factor in cytoadherence of P falciparum-parasitized erythrocytes (PEs), might also play a role in monocyte- and macrophage-mediated malarial clearance. Exposure of nonopsonized PEs to Fc receptor-blocked monocytes resulted in significant PE phagocytosis, accompanied by intense clustering of CD36 around the PEs. Phagocytosis was blocked 60% to 70% by monocyte pretreatment with monoclonal anti-CD36 antibodies but not by antibodies to alpha(v)beta(3), thrombospondin, intercellular adhesion molecule-1, or platelet/endothelial cell adhesion molecule-1. Antibody-induced CD36 cross-linking did result in the early increase of surface CD11b expression, but there was no increase in, or priming for, tumor necrosis factor (TNF)-alpha secretion following either CD36 cross linking or PE phagocytosis. CD36 clustering does support intracellular signaling: Antibody-induced cross-linking initiated intracellular tyrosine phosphorylation as well as extracellular signal-regulated kinase (ERK) and p38 mitogen-activated protein kinase (MAPK) phosphorylation. Both broad-spectrum tyrosine kinase inhibition (genistein) and selective ERK and p38 MAPK inhibition (PD98059 and SB203580, respectively) reduced PE uptake to almost the same extent as CD36 blockade. Thus, CD36-dependent binding and signaling appears to be crucial for the nonopsonic clearance of PEs and does not appear to contribute to the increase in TNF-alpha that is prognostic of poor outcome in clinical malaria. PMID- 11050009 TI - Regulation of hemoglobin synthesis and proliferation of differentiating erythroid cells by heme-regulated eIF-2alpha kinase. AB - Protein synthesis in reticulocytes depends on the availability of heme. In heme deficiency, inhibition of protein synthesis correlates with the activation of heme-regulated eIF-2alpha kinase (HRI), which blocks the initiation of protein synthesis by phosphorylating eIF-2alpha. HRI is a hemoprotein with 2 distinct heme-binding domains. Heme negatively regulates HRI activity by binding directly to HRI. To further study the physiological function of HRI, the wild-type (Wt) HRI and dominant-negative inactive mutants of HRI were expressed by retrovirus mediated transfer in both non-erythroid NIH 3T3 and mouse erythroleukemic (MEL) cells. Expression of Wt HRI in 3T3 cells resulted in the inhibition of protein synthesis, a loss of proliferation, and eventually cell death. Expression of the inactive HRI mutants had no apparent effect on the growth characteristics or morphology of NIH 3T3 cells. In contrast, expression of 3 dominant-negative inactive mutants of HRI in MEL cells resulted in increased hemoglobin production and increased proliferative capacity of these cells upon dimethyl-sulfoxide induction of erythroid differentiation. These results directly demonstrate the importance of HRI in the regulation of protein synthesis in immature erythroid cells and suggest a role of HRI in the regulation of the numbers of matured erythroid cells. PMID- 11050010 TI - Apoptosis in megaloblastic anemia occurs during DNA synthesis by a p53 independent, nucleoside-reversible mechanism. AB - Deficiency of folate or vitamin B(12) (cobalamin) causes megaloblastic anemia, a disease characterized by pancytopenia due to the excessive apoptosis of hematopoietic progenitor cells. Clinical and experimental studies of megaloblastic anemia have demonstrated an impairment of DNA synthesis and repair in hematopoietic cells that is manifested by an increased percentage of cells in the DNA synthesis phase (S phase) of the cell cycle, compared with normal hematopoietic cells. Both folate and cobalamin are required for normal de novo synthesis of thymidylate and purines. However, previous studies of impaired DNA synthesis and repair in megaloblastic anemia have concerned mainly the decreased intracellular levels of thymidylate and its effects on nucleotide pools and misincorporation of uracil into DNA. An in vitro model of folate-deficient erythropoiesis was used to study the relationship between the S-phase accumulation and apoptosis in megaloblastic anemia. The results indicate that folate-deficient erythroblasts accumulate in and undergo apoptosis in the S phase when compared with control erythroblasts. Both the S-phase accumulation and the apoptosis were induced by folate deficiency in erythroblasts from p53 null mice. The complete reversal of the S-phase accumulation and apoptosis in folate deficient erythroblasts required the exogenous provision of specific purines or purine nucleosides as well as thymidine. These results indicate that decreased de novo synthesis of purines plays as important a role as decreased de novo synthesis of thymidylate in the pathogenesis of megaloblastic anemia. PMID- 11050011 TI - Human ABC7 transporter: gene structure and mutation causing X-linked sideroblastic anemia with ataxia with disruption of cytosolic iron-sulfur protein maturation. AB - The human protein ABC7 belongs to the adenosine triphosphate-binding cassette transporter superfamily, and its yeast orthologue, Atm1p, plays a central role in the maturation of cytosolic iron-sulfur (Fe/S) cluster-containing proteins. Previously, a missense mutation in the human ABC7 gene was shown to be the defect in members of a family affected with X-linked sideroblastic anemia with cerebellar ataxia (XLSA/A). Here, the promoter region and the intron/exon structure of the human ABC7 gene were characterized, and the function of wild type and mutant ABC7 in cytosolic Fe/S protein maturation was analyzed. The gene contains 16 exons, all with intron/exon boundaries following the AG/GT rule. A single missense mutation was found in exon 10 of the ABC7 gene in 2 affected brothers with XLSA/A. The mutation was a G-to-A transition at nucleotide 1305 of the full-length cDNA, resulting in a charge inversion caused by the substitution of lysine for glutamate at residue 433 C-terminal to the putative sixth transmembrane domain of ABC7. Expression of normal ABC7 almost fully complemented the defect in the maturation of cytosolic Fe/S proteins in a yeast strain in which the ATM1 gene had been deleted (Deltaatm1 cells). Thus, ABC7 is a functional orthologue of Atm1p. In contrast, the expression of mutated ABC7 (E433K) or Atm1p (D398K) proteins in Deltaatm1 cells led to a low efficiency of cytosolic Fe/S protein maturation. These data demonstrate that both the molecular defect in XLSA/A and the impaired maturation of a cytosolic Fe/S protein result from an ABC7 mutation in the reported family. PMID- 11050012 TI - Retinoic acid stimulates erythropoietin gene transcription in embryonal carcinoma cells through the direct repeat of a steroid/thyroid hormone receptor response element half-site in the hypoxia-response enhancer. AB - We have previously reported that expression of the erythropoietin (Epo) gene in mouse embryonal cells was not induced by hypoxia, although hypoxia induced other hypoxia-inducible genes. This study identifies retinoic acid (RA) as an inducer for Epo production in the embryonal carcinoma cell lines P19 and F9. RA induced Epo production through the transcriptional activation of the Epo gene in an oxygen-independent manner. With the use of reporter assays in P19 cells, it is shown that a direct repeat of the nuclear hormone receptor-binding motif separated by a 2-bp spacer (DR-2) in the hypoxia-response enhancer was responsible for the transcriptional activation by RA. Electrophoretic mobility shift assays show that nuclear extracts from P19 cells contained RA receptor complexes that bound to DR-2. In human hepatoma Hep3B cells, an orphan receptor, hepatocyte nuclear factor-4, strongly augmented hypoxic induction of the Epo gene in cooperation with hypoxia-inducible factor-1 (HIF-1) by binding to DR-2, whereas in P19 cells, the interaction of RA receptors with DR-2 was sufficient for RA-induced transcriptional activation of the Epo gene without the requirement of the HIF-1 site. These results suggest that DR-2 regulates expression of the Epo gene by acting as the binding site for different transcription factors in different types of cells. PMID- 11050013 TI - Assessment of bone marrow stem cell reserve and function and stromal cell function in patients with autoimmune cytopenias. AB - To investigate whether bone marrow (BM) stem cell compartment and/or BM microenvironment are affected by the immune insult in autoimmune cytopenias (AICs), BM stem cell reserve and function and BM stromal function were studied in 15 AIC patients. Stem cells were evaluated by means of flow cytometry, clonogenic progenitor cell assays, long-term BM cultures (LTBMCs), and limiting dilution assay for quantification of long-term-culture initiating cells (LTC-ICs). Stromal cell function was assessed with the use of preformed irradiated LTBMCs from patients and normal controls, recharged with normal CD34(+) cells. AIC patients exhibited a high number of CD34(+), CD34(+)/CD38(+), and CD34(+)/CD38(-) cells; high frequency of granulocyte-macrophage colony forming units in the BM mononuclear cell fraction; high colony recovery in LTBMCs; and normal LTC-IC frequency. Patient BM stromal layers displayed normal hematopoietic-supporting capacity and increased production of granulocyte-colony stimulating factor. Data from this study support the concept that AIC patients with severe, resistant disease might be appropriate candidates for autologous stem cell transplantation. PMID- 11050014 TI - Secretory phospholipase A(2) predicts impending acute chest syndrome in sickle cell disease. AB - Acute chest syndrome (ACS) is the leading cause of death in sickle cell disease. Severe ACS often develops in the course of a vaso-occlusive crisis (VOC), but currently there are no predictors for its development. Secretory phospholipase A(2) (sPLA(2)), a potent inflammatory mediator, is elevated in ACS, and previous work suggests that sPLA(2) predicts impending ACS. We prospectively evaluated sPLA(2) concentration during 21 admissions for VOC; 6 of these patients went on to develop ACS. Elevation of sPLA(2) was detected all 6 patients 24 to 48 hours before ACS was clinically diagnosed. Adding the requirement for fever raised the specificity of sPLA(2) to 87% while retaining 100% sensitivity. These data indicate that sPLA(2) can be useful in alerting the clinician to patients with impending ACS. In addition, sPLA(2) may be useful for instituting early therapies to prevent or reduce the clinical morbidity of ACS. PMID- 11050015 TI - Molecular evidence of organ-related transmission of Kaposi sarcoma-associated herpesvirus or human herpesvirus-8 in transplant patients. AB - In transplant patients, Kaposi sarcoma (KS)-associated herpesvirus or human herpesvirus-8 (HHV-8) infection is associated with the development of KS, primary effusion lymphoma and Castleman disease. Whether HHV-8 is either reactivated in the recipient or transmitted by the donor has been investigated so far only by serologic studies. Thus, we addressed the issue of HHV-8 transmission in the transplantation setting by molecular methods. We exploited the high level variability of the orf-K1 gene and the polymorphism of the orf-73 gene of the HHV 8 genome to assess the genetic relatedness of the HHV-8 strains identified in the posttransplant KS lesions that developed, simultaneously, 20 months after transplantation, in 2 recipients of twin kidneys from the same cadaver donor. The 100% identity of nucleotide sequence of the most variable viral region and the presence of the same, single orf-73 type in both patients provides strong molecular evidence of organ-related transmission of HHV-8 in the setting of transplantation. PMID- 11050016 TI - Adhesion to 90K (Mac-2 BP) as a mechanism for lymphoma drug resistance in vivo. AB - A cell-adhesive protein of the human serum, 90K binds galactin-3, beta1 integrins, collagens, and fibronectin, and it is of importance in cell-cell and cell-extracellular matrix adhesion. Serum 90K levels in 137 patients with lymphoma were measured by enzyme-linked immunosorbent assay. Compared with healthy controls, pretreatment serum 90K levels in patients with lymphoma were elevated (P <.001). Of 97 patients who showed objective response to treatment, 20 (21%) had pretreatment 90K levels above the normal cutoff compared with 17 (53%) of 32 patients who did not respond (P =.002). When used as a plastic-immobilized substrate, 90K caused a significant reduction in chemotherapy-induced apoptosis of Jurkat T lymphoma cells. This finding could explain the lack of response in lymphoma patients with high 90K serum levels. PMID- 11050017 TI - Drug-resistant human cytomegalovirus infection in children after allogeneic stem cell transplantation may have different clinical outcomes. AB - Three seropositive pediatric recipients of allogeneic stem cell transplantation out of a group of 42 patients receiving T-cell-depleted, unrelated transplants and 37 patients receiving T-cell-depleted, haploidentical transplants were monitored longitudinally for human cytomegalovirus (HCMV) infection and the emergence of antiviral drug resistance. Early in the posttransplant course, all 3 patients developed HCMV mutations conferring drug resistance to ganciclovir. One child additionally developed multidrug resistance to foscarnet and cidofovir, with mutations in the viral phosphotransferase gene (UL97) and the DNA-polymerase gene (UL54) being found. These data show that resistant HCMV infection does not necessarily correlate with a severe clinical outcome. The early detection of genotypic resistance up to 129 days before the emergence of phenotypic resistance and the dissociation of resistance patterns among different body sites emphasize the importance of genotypic analyses of different DNA specimens for an efficient antiviral therapy. T-cell-depleted children having transplantation might be at an increased risk for the development of drug resistance. PMID- 11050018 TI - Low B-cell and monocyte counts on day 80 are associated with high infection rates between days 100 and 365 after allogeneic marrow transplantation. AB - To ascertain which mononuclear cell subset deficiency plays a role in the marrow transplant recipient's susceptibility to infections, mononuclear cell subset counts were prospectively determined in 108 patients on day 80. Infections occurring between day 100 and 365 were recorded by an investigator blinded to the subset counts. In univariate analyses, the counts of the following subsets showed a significant inverse correlation with infection rates: total B cells, IgD(+) B cells, IgD(-) B cells, total CD4 T cells, CD28(+) CD4 T cells, CD28(-) CD4 T cells, CD45RA(low/-) CD4 T cells and monocytes. In multivariate analyses, the counts of the following subsets remained significantly inversely correlated with the infection rates: total B cells (P =.0004) and monocytes (P =.009). CD28(-) CD8 T-cell counts showed no correlation with infection rates. In conclusion, the susceptibility of patients to infections late posttransplant may be due in part to the slow reconstitution of B cells and monocytes. PMID- 11050019 TI - MRI: measure of efficacy. PMID- 11050020 TI - Characterization of the decision-making deficit of patients with ventromedial prefrontal cortex lesions. AB - On a gambling task that models real-life decisions, patients with bilateral lesions of the ventromedial prefrontal cortex (VM) opt for choices that yield high immediate gains in spite of higher future losses. In this study, we addressed three possibilities that may account for this behaviour: (i) hypersensitivity to reward; (ii) insensitivity to punishment; and (iii) insensitivity to future consequences, such that behaviour is always guided by immediate prospects. For this purpose, we designed a variant of the original gambling task in which the advantageous decks yielded high immediate punishment but even higher future reward. The disadvantageous decks yielded low immediate punishment but even lower future reward. We measured the skin conductance responses (SCRs) of subjects after they had received a reward or punishment. Patients with VM lesions opted for the disadvantageous decks in both the original and variant versions of the gambling task. The SCRs of VM lesion patients after they had received a reward or punishment were not significantly different from those of controls. In a second experiment, we investigated whether increasing the delayed punishment in the disadvantageous decks of the original task or decreasing the delayed reward in the disadvantageous decks of the variant task would shift the behaviour of VM lesion patients towards an advantageous strategy. Both manipulations failed to shift the behaviour of VM lesion patients away from the disadvantageous decks. These results suggest that patients with VM lesions are insensitive to future consequences, positive or negative, and are primarily guided by immediate prospects. This 'myopia for the future' in VM lesion patients persists in the face of severe adverse consequences, i.e. rising future punishment or declining future reward. PMID- 11050021 TI - The functional neuroanatomy of social behaviour: changes in cerebral blood flow when people with autistic disorder process facial expressions. AB - Although high-functioning individuals with autistic disorder (i.e. autism and Asperger syndrome) are of normal intelligence, they have life-long abnormalities in social communication and emotional behaviour. However, the biological basis of social difficulties in autism is poorly understood. Facial expressions help shape behaviour, and we investigated if high-functioning people with autistic disorder show neurobiological differences from controls when processing emotional facial expressions. We used functional MRI to investigate brain activity in nine adults with autistic disorder (mean age +/- standard deviation 37 +/- 7 years; IQ 102 +/ 15) and nine controls (27 +/- 7 years; IQ 116 +/- 10) when explicitly (consciously) and implicitly (unconsciously) processing emotional facial expressions. Subjects with autistic disorder differed significantly from controls in the activity of cerebellar, mesolimbic and temporal lobe cortical regions of the brain when processing facial expressions. Notably, they did not activate a cortical 'face area' when explicitly appraising expressions, or the left amygdala region and left cerebellum when implicitly processing emotional facial expressions. High-functioning people with autistic disorder have biological differences from controls when consciously and unconsciously processing facial emotions, and these differences are most likely to be neurodevelopmental in origin. This may account for some of the abnormalities in social behaviour associated with autism. PMID- 11050022 TI - Upper and lower face apraxia: role of the right hemisphere. AB - The aim of this study was to evaluate face apraxia in left- and right-hemisphere damaged patients both in the acute and chronic stage of their disease. Two newly devised tests that assess movements of the upper and lower face districts were employed. On the whole, the proportion of left-hemisphere-damaged patients showing face apraxia were 46 and 68% for upper and lower face, respectively. A substantial proportion of right-hemisphere-damaged patients also showed face apraxia, i.e. 44% upper face and 38% lower face. Concomitant variables such as general severity, locus of lesion, language or visuo-spatial impairments, presence of neglect, interval from stroke, peculiarity of clusters of items or scoring procedures did not account for these results. These findings suggest that face apraxia in some patients may affect movements of the upper face district and that the right hemisphere plays a significant part in both upper and lower face praxis. PMID- 11050023 TI - Motor cortex excitability in stiff-person syndrome. AB - Muscle stiffness in stiff-person syndrome (SPS) is produced by continuous, involuntary firing of motor units that is thought to be caused by an autoimmune mediated dysfunction of GABA-ergic inhibitory neurones. We have postulated that the loss of GABA-ergic inputs from spinal interneurones alone is insufficient to produce tonic firing of motor neurones and that excessive supraspinal excitation could also play a role. To determine whether SPS is associated with dysfunction in supraspinal GABA-ergic neurones, we assessed the excitability of the motor cortex with transcranial magnetic stimulation (TMS) in seven SPS patients and seven age-matched healthy volunteers. SPS patients had normal central motor conduction times, normal thresholds for motor evoked potentials (MEPs) in leg muscles, and a normal MEP stimulus versus response recruitment curve with increasing TMS intensities in resting hand and leg muscles. Cortical silent periods were shortened in leg muscles. Intracortical inhibition and excitation were assessed while recording from the abductor pollicis brevis, using a paired pulse TMS paradigm with subthreshold conditioning stimuli. Patients had decreased inhibition and markedly increased facilitation at short intervals. Using paired suprathreshold TMS, patients exhibited increased facilitation at 20- and 40-ms intervals. These results point to a hyperexcitability of the motor cortex in SPS, which could be explained by impairment of supraspinal GABA-ergic neurones, leading to an impaired balance between inhibitory and excitatory intracortical circuitry. PMID- 11050024 TI - Understanding dissociations in dyscalculia: a brain imaging study of the impact of number size on the cerebral networks for exact and approximate calculation. AB - Neuropsychological studies have revealed different subtypes of dyscalculia, including dissociations between exact calculation and approximation abilities, and an impact of number size on performance. To understand the origins of these effects, we measured cerebral activity with functional MRI at 3 Tesla and event related potentials while healthy volunteers performed exact and approximate calculation tasks with small and large numbers. Bilateral intraparietal, precentral, dorsolateral and superior prefrontal regions showed greater activation during approximation, while the left inferior prefrontal cortex and the bilateral angular regions were more activated during exact calculation. Increasing number size during exact calculation led to increased activation in the same bilateral intraparietal regions as during approximation, as well the left inferior and superior frontal gyri. Event-related potentials gave access to the temporal dynamics of calculation processes, showing that effects of task and of number size could be found as early as 200-300 ms following problem presentation. Altogether, the results reveal two cerebral networks for number processing. Rote arithmetic operations with small numbers have a greater reliance on left-lateralized regions, presumably encoding numbers in verbal format. Approximation and exact calculation with large numbers, however, put heavier emphasis on the left and right parietal cortices, which may encode numbers in a non-verbal quantity format. Subtypes of dyscalculia can be explained by lesions disproportionately affecting only one of these networks. PMID- 11050025 TI - The effect of interferon beta-1b treatment on MRI measures of cerebral atrophy in secondary progressive multiple sclerosis. European Study Group on Interferon beta 1b in secondary progressive multiple sclerosis. AB - The recently completed European trial of interferon beta-1b (IFNbeta-1b) in patients with secondary progressive multiple sclerosis (SP multiple sclerosis) has given an opportunity to assess the impact of treatment on cerebral atrophy using serial MRI. Unenhanced T(1)-weighted brain imaging was acquired in a subgroup of 95 patients from five of the European centres; imaging was performed at 6-month intervals from month 0 to month 36. A blinded observer measured cerebral volume on four contiguous 5 mm cerebral hemisphere slices at each time point, using an algorithm with a high level of reproducibility and automation. There was a significant and progressive reduction in cerebral volume in both placebo and treated groups, with a mean reduction of 3.9 and 2.9%, respectively, by month 36 (P = 0.34 between groups). Exploratory subgroup analyses indicated that patients without gadolinium (Gd) enhancement at the baseline had a greater reduction of cerebral volume in the placebo group (mean reduction at month 36: placebo 5.1%, IFNbeta-1b 1.8%, P < 0.05) whereas those with Gd-enhancing lesions showed a trend to greater reduction of cerebral volume if the patient was on IFNbeta-1b (placebo 2.6%, IFNbeta-1b 3.7%; P > 0.05). These results are consistent with ongoing tissue loss in both arms of this study of secondary progressive multiple sclerosis. This finding is concordant with previous observations that disease progression, although delayed, is not halted by IFNbeta. The different pattern seen in patients with and without baseline gadolinium enhancement suggests that part of the cerebral volume reduction observed in IFNbeta-treated patients may be due to the anti inflammatory/antioedematous effect of the drug. Longer periods of observation and larger groups of patients may be needed to detect the effects of treatment on cerebral atrophy in this population of patients with advanced disease. PMID- 11050026 TI - Task-related changes of transmission in the pathway of heteronymous spinal recurrent inhibition from soleus to quadriceps motor neurones in man. AB - An H reflex conditioning technique was used to monitor the transmission of heteronymous recurrent inhibition from soleus to quadriceps motor neurones of the human lower limb. Inhibition declined during quadriceps muscle contraction under all conditions examined, falling to zero at around one-third of the maximum voluntary contraction. Inhibition declined during soleus muscle contraction in sitting, standing and bicycling tasks. The level of inhibition assessed at a given (weaker than 30%) level of quadriceps contraction was reduced during postural tasks involving quadriceps and soleus co-contraction (standing and late stance phase of walking) when compared with sitting and performing matched voluntary muscle contractions. The level of inhibition during the mid-power stroke of a bicycling task, which also involved co-contraction of quadriceps and soleus, was greater than during matched voluntary muscle contractions while sitting. It is concluded that the pathway of heteronymous recurrent inhibition from soleus to quadriceps motor neurones is under at least two types of control: one related to the task, which sets the operating range, and a second which couples inhibition to the level of muscle contraction. Multiple control pathways are consistent with the diverse effects on recurrent inhibition reported in subjects with upper motor neurone lesions. PMID- 11050027 TI - The neurology of saccades and covert shifts in spatial attention: an event related fMRI study. AB - Visual neglect occurs most frequently and persistently after lesions that include the right supramarginal gyrus (SMG), a part of the inferior parietal lobule. Patients with this syndrome make very few saccades to the left, and show abnormal performance on tasks in which they must covertly shift their attention to the left, suggesting that the right SMG is involved in the generation of saccades and attention shifts. Functional imaging studies of saccades and covert attention shifts in the normal brain, however, have shown weak or absent responses in both SMGs. We used event-related functional MRI to re-examine the responses to saccades and attention shifts within a single experiment, and to assess responses to left- and right-sided stimuli independently. When subjects made saccades to peripheral stimuli, the expected responses were seen in striate and prestriate cortex, the superior parietal lobules, the frontal eye fields, the supplementary motor area and the anterior insulae. In addition there was a response in the right SMG but not in the left SMG, as predicted from the clinical literature. When subjects made a covert visual assessment of the peripheral stimulus without any saccade, greater activity was seen in all of the areas in the frontoparietal network. Each area showed a bias towards contralateral stimuli, with two exceptions: the anterior insulae gave mainly ipsilateral responses, whilst the right SMG gave equal responses to right- and left-sided stimuli. These findings are discussed in the context of current theories pertaining to the clinical syndrome of neglect. PMID- 11050028 TI - Extramotor involvement in ALS: PET studies with the GABA(A) ligand [(11)C]flumazenil. AB - We used the benzodiazepine GABA(A) marker [(11)C] flumazenil to study cerebral dysfunction in amyotrophic lateral sclerosis (ALS) with PET. Seventeen non demented patients with clinically definite or probable ALS were scanned and statistical parametric maps were derived to localize changes in regional flumazenil volumes of distribution (FMZVD), which correlate closely with receptor density (B(max)), and the results were compared with those of 17 controls. The ALS group showed statistically significant decreases in relative FMZVD in the prefrontal cortex (areas 9 and 10 bilaterally), parietal cortex (area 7 bilaterally), visual association cortex (area 18 bilaterally) and left motor/premotor cortex (including area 4) (P < 0.001). Relative reductions in FMZVD were also seen in the left ventrolateral and dorsolateral prefrontal cortex (areas 45, 46 and 47), Broca's area and the right temporal (area 21) and right visual association cortex (area 19). These observations suggest that cerebral dysfunction in ALS involves motor/premotor and extramotor areas, particularly the prefrontal regions. PMID- 11050029 TI - Dyskinesias and motor fluctuations in Parkinson's disease. A community-based study. AB - We investigated the prevalence of dyskinesias and motor fluctuations, and the factors determining their occurrence, in a community-based population of patients with Parkinson's disease. Among 124 patients with Parkinson's disease, 87 (70%) had received a levodopa preparation. Among these 87 patients, 28% were experiencing treatment-induced dyskinesias and 40% response fluctuations. The prevalence of motor fluctuations was best predicted by disease duration and dose of levodopa, whereas dyskinesias could be best predicted by duration of treatment. Patients with a shorter time from symptom onset to initiation of levodopa and younger patients had developed motor complications earlier, and patients who had started treatment with a dopamine agonist had developed these treatment complications later. Although a satisfactory response to medication was associated with higher rates of motor complications, poor or moderate response was associated with lower quality of life in patients with a disease duration of /=10 years. We conclude that motor fluctuations are most strongly related to disease duration and dose of levodopa, and dyskinesias to duration of levodopa treatment. However, poorer quality of life associated with inadequate dosage of levodopa may be the price for a low rate of motor complications in patients with Parkinson's disease. PMID- 11050030 TI - Neural representations of skilled movement. AB - The frontal and parietal cortex are intimately involved in the representation of goal-directed movements, but the crucial neuroanatomical sites are not well established in humans. In order to identify these sites more precisely, we studied stroke patients who had the classic syndrome of ideomotor limb apraxia, which disrupts goal-directed movements, such as writing or brushing teeth. Patients with and without limb apraxia were identified by assessing errors imitating gestures and specifying a cut-off for apraxia relative to a normal control group. We then used MRI or CT for lesion localization and compared areas of overlap in those patients with and without limb apraxia. Patients with ideomotor limb apraxia had damage lateralized to a left hemispheric network involving the middle frontal gyrus and intraparietal sulcus region. Thus, the results revealed that discrete areas in the left hemisphere of humans are critical for control of complex goal-directed movements. PMID- 11050031 TI - Evidence for adaptive functional changes in the cerebral cortex with axonal injury from multiple sclerosis. AB - Axonal injury occurs even in the earliest stages of multiple sclerosis. Magnetic resonance spectroscopic imaging (MRSI) measurements of brain N:-acetylaspartate (NAA), a marker of axonal integrity, show that this axonal injury can occur even in the absence of clinically evident functional impairments. To test whether cortical adaptive responses contribute to the maintenance of normal motor function in patients with multiple sclerosis, we performed MRSI and functional MRI (fMRI) examinations of nine multiple sclerosis patients who had unimpaired hand function. We found that activation of the ipsilateral sensorimotor cortex with simple hand movements was increased by a mean of fivefold relative to normal controls (n = 8) and that the extent of this increase was strongly correlated (sigma = -0.93, P = 0.001) with decreases in brain NAA. These results suggest that compensatory cortical adaptive responses may help to account for the limited relationship between conventional MRI measures of lesion burden and clinical measures of disability, and that therapies directed towards promoting cortical reorganization in response to brain injury could enhance recovery from relapses of multiple sclerosis. PMID- 11050032 TI - The peripheral benzodiazepine binding site in the brain in multiple sclerosis: quantitative in vivo imaging of microglia as a measure of disease activity. AB - This study identifies by microautoradiography activated microglia/macrophages as the main cell type expressing the peripheral benzodiazepine binding site (PBBS) at sites of active CNS pathology. Quantitative measurements of PBBS expression in vivo obtained by PET and [(11)C](R)-PK11195 are shown to correspond to animal experimental and human post-mortem data on the distribution pattern of activated microglia in inflammatory brain disease. Film autoradiography with [(3)H](R) PK11195, a specific ligand for the PBBS, showed minimal binding in normal control CNS, whereas maximal binding to mononuclear cells was found in multiple sclerosis plaques. However, there was also significantly increased [(3)H](R)-PK11195 binding on activated microglia outside the histopathologically defined borders of multiple sclerosis plaques and in areas, such as the cerebral central grey matter, that are not normally reported as sites of pathology in multiple sclerosis. A similar pattern of [(3)H](R)-PK11195 binding in areas containing activated microglia was seen in the CNS of animals with experimental allergic encephalomyelitis (EAE). In areas without identifiable focal pathology, immunocytochemical staining combined with high-resolution emulsion autoradiography demonstrated that the cellular source of [(3)H](R)-PK11195 binding is activated microglia, which frequently retains a ramified morphology. Furthermore, in vitro radioligand binding studies confirmed that microglial activation leads to a rise in the number of PBBS and not a change in binding affinity. Quantitative [(11)C](R)-PK11195 PET in multiple sclerosis patients demonstrated increased PBBS expression in areas of focal pathology identified by T(1)- and T(2)-weighted MRI and, importantly, also in normal-appearing anatomical structures, including cerebral central grey matter. The additional binding frequently delineated neuronal projection areas, such as the lateral geniculate bodies in patients with a history of optic neuritis. In summary, [(11)C](R) PK11195 PET provides a cellular marker of disease activity in vivo in the human brain. PMID- 11050033 TI - Cortical activation patterns of affective speech processing depend on concurrent demands on the subvocal rehearsal system. A DC-potential study. AB - In order to delineate brain regions specifically involved in the processing of affective components of spoken language (affective or emotive prosody), we conducted two event-related potential experiments. Cortical activation patterns were assessed by recordings of direct current components of the EEG signal from the scalp. Right-handed subjects discriminated pairs of declarative sentences with either happy, sad or neutral intonation. Each stimulus pair was derived from two identical original utterances that, due to digital signal manipulations, slightly differed in fundamental frequency (F0) range or in duration of stressed syllables. In the first experiment, subjects were asked: (i) to denote the original emotional category of each sentence pair and (ii) to decide which of the two items displayed stronger emotional expressiveness. Participants in the second experiment were asked to repeat the utterances using inner speech during stimulus presentation in addition to the discrimination task. In the absence of inner speech, a predominant activation of right frontal regions was observed, irrespective of emotional category. In the second experiment, a bilateral activation with left frontal preponderance emerged from discrimination during additional performance of inner speech. Compared with the first experiment, a new pattern of acoustic signal processing arose. A relative decrease of brain activity during processing of F0 stimulus variants was observed together with increased activation during discrimination of duration-manipulated sentence pairs. Analysis of behavioural data revealed no significant differences in evaluation of expressiveness between the two experiments. We conclude that the topographical shift of cortical activity originates from left hemisphere (LH) mechanisms of speech processing that centre around the subvocal rehearsal system as an articulatory control component of the phonological loop. A strong coupling of acoustic input and (planned) verbal output channel in the LH is initiated by subvocal articulatory activity like inner speech. These neural networks may provide interpretations of verbal acoustic signals in terms of motor programs and facilitate continuous control of speech output by comparing the signal produced with that intended. Most likely, information on motor aspects of suprasegmental signal characteristics contributes to the evaluation of affective components of spoken language. In consequence, the right hemisphere (RH) holds a merely relative dominance, both for processing of F0 and for evaluation of emotional significance of sensory input. Psychophysically, an important determinant on expression of lateralization patterns seems to be given by the degree of communicative demands such as solely perceptive (RH) or perceptive and verbal expressive (RH and LH). PMID- 11050034 TI - Left tactile extinction following visual stimulation of a rubber hand. AB - In close analogy with neurophysiological findings in monkeys, neuropsychological studies have shown that the human brain constructs visual maps of space surrounding different body parts. In right-brain-damaged patients with tactile extinction, the existence of a visual peripersonal space centred on the hand has been demonstrated by showing that cross-modal visual-tactile extinction is segregated mainly in the space near the hand. That is, tactile stimuli on the contralesional hand are extinguished more consistently by visual stimuli presented near the ipsilesional hand than those presented far from it. Here, we report the first evidence in humans that this hand-centred visual peripersonal space can be coded in relation to a seen rubber replica of the hand, as if it were a real hand. In patients with left tactile extinction, a visual stimulus presented near a seen right rubber hand induced strong cross-modal visual-tactile extinction, similar to that obtained by presenting the same visual stimulus near the patient's right hand. Critically, this specific cross-modal effect was evident when subjects saw the rubber hand as having a plausible posture relative to their own body (i.e. when it was aligned with the subject's right shoulder). In contrast, cross-modal extinction was strongly reduced when the seen rubber hand was arranged in an implausible posture (i. e. misaligned with respect to the subject's right shoulder). We suggest that this phenomenon is due to the dominance of vision over proprioception: the system coding peripersonal space can be 'deceived' by the vision of a fake hand, provided that its appearance looks plausible with respect to the subject's body. PMID- 11050035 TI - Hepatitis C virus resistance to antiviral therapy. PMID- 11050036 TI - Silymarin in the treatment of patients with primary biliary cirrhosis with a suboptimal response to ursodeoxycholic acid. AB - Ursodeoxycholic acid (UDCA) is a safe and effective medical therapy for most patients with primary biliary cirrhosis (PBC), but some patients show an incomplete response. Silymarin is a potent antioxidant with immunomodulatory and antifibrotic properties. The aim of this study was to evaluate the safety and assess the efficacy of silymarin in patients with PBC who had shown a suboptimal response to UDCA. Twenty-seven patients with PBC who had been on UDCA (13-15 mg/kg/day) therapy for 7 to 221 months and had shown a persistent elevation of alkaline phosphatase activity at least 2 times the upper limit of normal for more than 6 months were enrolled. Oral silymarin, 140 mg 3 times daily was given for 1 year, and patients continued on the same dosage of UDCA. No significant changes in serum alkaline phosphatase activity (897 +/- 84 vs. 876 +/- 95, P =.5), total bilirubin (0.9 +/- 0.1 vs. 1 +/- 0.1, P =.07), aspartate transaminase (AST) (58 +/- 5 vs. 56 +/- 6, P =.4), albumin (4.0 +/-.06 vs. 4.1 +/-.06, P =.4), or Mayo risk score (3.82 +/- 0.2 vs. 3.88 +/- 0.2, P =.4) were noted after 1 year of treatment with combination therapy. Transitory gastrointestinal adverse events occurred in 2 patients. In conclusion, although silymarin was well tolerated, this medication did not provide benefit to patients with PBC responding suboptimally to UDCA. The results of this pilot study would seem to discourage further controlled trials of silymarin in patients with PBC. PMID- 11050037 TI - Fine specificity of T cells reactive to human PDC-E2 163-176 peptide, the immunodominant autoantigen in primary biliary cirrhosis: implications for molecular mimicry and cross-recognition among mitochondrial autoantigens. AB - The anti-mitochondrial antibody response in primary biliary cirrhosis (PBC) is primarily directed at E2 components of PDC, OGDC, and BCOADC, and E3BP. Previous work has shown that the immunodominant autoreactive T- cell epitope is the PDC-E2 163-176 peptide, restricted by HLA DR53. To address molecular mimicry and cross recognition among mitochondrial autoantigens, we analyzed reactivity, including agonism and antagonism assays, to a series of single amino acid-substituted peptides using cloned T-cell lines in PBC and controls. Interestingly, fine specificities were unique for every single T-cell clone, but the clones could be categorized into two distinct groups based on recognition motifs of the T-cell receptor (TCR) ligand: group A (170)ExDK(173) and group B (168)EIExD(172). (170)E is the most critical TCR contact residue for both groups of cloned T-cell lines, whereas (173)K and (168)E are the critical TCR contact residues for group A and group B cloned T-cell lines, respectively. More importantly, some group A-cloned T-cell lines cross-reacted to human E3BP 34-47, human OGDC-E2 100-113, and several peptides derived from various microbial proteins carrying an ExDK motif, whereas group B-cloned T-cell lines reacted only to E3BP 34-47 carrying an EIExD motif. Furthermore, an RGxG motif was exclusively found in the complementarity determining region (CDR3) of the TCR Vbeta in the group B-cloned T-cell lines, while G, S, and/or R were frequently found in the CDR3 of the TCR Vbeta in the group A-cloned T-cell lines. These data provide a framework for understanding molecular mimicry among mitochondrial antigens. PMID- 11050038 TI - Mucosal immunity and primary biliary cirrhosis: presence of antimitochondrial antibodies in urine. AB - We have shown that IgA-class antimitochondrial autoantibodies (AMA) can be detected in the bile and saliva of patients with PBC, suggesting that AMA are secreted into the luminal fluid across bile ducts and salivary glands. These data prompted us to determine whether AMA of the IgA isotype may be transported across other epithelial mucosa. Therefore, we tested for the presence of AMA in the urine specimens of 83 patients with PBC and 58 non-PBC controls including healthy individuals and patients with other liver diseases. Patients enrolled in this study had no history of renal disease, and we confirmed there was less than 50 microgram/mL of protein in each of the urine specimens. Interestingly, we found that AMA were present in the urine of 71/83 (86%) of all patients with PBC and in 71/78 (91%) of patients with PBC that were serum AMA positive. In contrast, AMA were not detected in any of the 58 control urine specimens. Of particular interest, AMA of the IgA isotype was present in 57/83 (69%) of patients with PBC, and in 52 of these 57, we found secretory-type IgA. In a nested random subgroup of urine samples, the prevalence of the IgA2 AMA was 6/18 (33%), significantly lower than in matched serum samples, 13/16 (81%, P =.007). These data show that AMA of the IgA isotype is secreted into urine from the uroepithelium of patients with PBC, and support the thesis that PBC originated from either a mucosal challenge or a loss of mucosal tolerance. PMID- 11050039 TI - A study of carboplatin-coated tube for the unresectable cholangiocarcinoma. AB - Most cases of cholangiocarcinoma have reached an unresectable stage by the time they are discovered despite significant progress of diagnostic modalities. Many of these patients with obstructive jaundice are often treated by biliary drainage using stents to relieve the jaundice. However, the stent patency period is as short as 3 to 9 months because of tumor ingrowth or overgrowth, and mean survival is at most 12 months. Therefore, both continuous relief of obstructive jaundice and local control of the tumor are required in the treatment for advanced cholangiocarcinoma. In this investigation, we developed a new percutaneous transhepatic biliary drainage tube coated with carboplatin (carboplatin-coated tube; CCT). CCT continuously released a fixed amount of carboplatin for 4 weeks and showed an antitumor effect on human cholangiocarcinoma cell line HuCC-T1 in vitro. When CCT was embedded in subcutaneous tumor inoculated in nude mice, a significant reduction of tumor size with no apparent damage to normal adjacent tissue was observed. On the basis of these studies, 5 patients with inoperable cholangiocarcinoma were treated with CCT for 4 weeks. Overall efficacy rate of 5 patients with cholangiocarcinoma was 60% (partial response in 3 and no change in 2). No apparent side effect was observed in these patients. Thus, CCT may provide a new treatment modality for this disease. Randomized controlled trials comparing CCT therapy with palliative stenting are required to confirm these results. PMID- 11050040 TI - Reliability and validity of the NIDDK-QA instrument in the assessment of quality of life in ambulatory patients with cholestatic liver disease. AB - The NIDDK-QA instrument, developed and widely used in liver transplant recipients, assesses quality of life (QOL) in four domains, including liver disease symptoms, physical function, health satisfaction, and overall well-being. We investigated whether the instrument may be used as a disease-specific instrument in ambulatory patients with cholestatic liver disease. The NIDDK-QA instrument was administered in 96 patients with primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC) seen at the Mayo Clinic. The SF-36, a well-established generic instrument, was also administered. Standard measures for test-retest reliability, internal consistency, and discriminant and concurrent validity were examined. All patients were ambulatory with mostly normal levels of serum bilirubin and albumin concentrations. The reliability of the NIDDK-QA, as measured by test-retest correlation (Pearson coefficients: 0.82-0.99, P <.01) and by internal consistency (Cronbach's alpha: 0.87-0.94) exceeded conventional acceptability criteria. The correlation between domain scores of the NIDDK-QA and SF-36 was clear and logical in that the physical function domain of NIDDK-QA strongly correlated with the physical component summary score of SF-36 (r = 0.86, P <.01). The overall well-being domain of the NIDDK-QA was closely associated with the mental summary score of SF-36 (r = 0.69, P <.01). Among PBC patients, there was a modest yet significant correlation between the Mayo risk score and overall well-being (r = -0.26, P =.03). In the assessment of QOL in patients with cholestatic liver disease, NIDDK-QA is found reliable and valid. These data, combined with our previous study, demonstrate its applicability in a wide spectrum of disease severity, ranging from early, ambulatory-phase disease to decompensated cirrhosis necessitating liver transplantation. PMID- 11050041 TI - The hemodynamic response to medical treatment of portal hypertension as a predictor of clinical effectiveness in the primary prophylaxis of variceal bleeding in cirrhosis. AB - In the prevention of variceal rebleeding, it is already established that hemodynamic response to drug treatment (decrease in hepatic venous pressure gradient [HVPG] to 12 mm Hg or by >20%) is predictive of clinical effectiveness. In primary prophylaxis very few clinical data are available. We assessed the role of the hemodynamic response to beta-blockers or beta-blockers plus nitrates in predicting clinical efficacy of prophylaxis. A total of 49 cirrhotic patients with varices at risk of bleeding, without prior variceal bleeding, were investigated by hepatic vein catheterization before and after 1 to 3 months of chronic treatment with nadolol or nadolol plus isosorbide mononitrate, and were followed during treatment for up to 5 years. A total of 30 patients (61%) were good hemodynamic responders, and among them in 12 (24%) HVPG was 3 x the upper limit of normal [ULN]) was associated with a more rapid selection of drug-resistant mutants (P =.027) and a high hepatitis B virus (HBV) DNA level (>1,497 Meq/mL, bDNA) with a more rapid occurrence of phenotypic resistance (P =.04). At the time of viral breakthrough, the mean serum HBV-DNA values were not different from the pretreatment values (P =.37). ALT levels were higher in anti HBe-positive patients compared with pretreatment values and to HBeAg-positive patients (P =.01). In 8 patients, antiviral therapy was modified after viral breakthrough, with the introduction of famciclovir and/or interferon alfa. Viral DNA became undetectable by bDNA in 3 patients who received interferon. Our results suggest that genotypic assays for polymerase mutant detection and quantitative determination of viremia with highly sensitive assay are warranted for an optimal monitoring of antiviral therapy of chronic hepatitis B. PMID- 11050060 TI - Intrahepatic mRNA expression of interferon-inducible antiviral genes in liver diseases: dsRNA-dependent protein kinase overexpression and RNase L inhibitor suppression in chronic hepatitis C. AB - As a part of the defense mechanism of the host to viral infection, interferons induce the transcription of several genes. These interferon-inducible genes contribute to the eradication of the viruses. Whereas some studies suggested the participation of a dsRNA-dependent protein kinase in the host reaction to hepatitis C virus infection, the involvement of other interferon-inducible genes has not been evaluated. Furthermore, there has been no analysis on the expression profile of multiple interferon-inducible genes. The aim of this study was to clarify the hepatic mRNA expression profile of interferon-inducible genes with a special concern to chronic hepatitis C. A total of 76 liver biopsy samples (28 with chronic hepatitis C, 10 with chronic hepatitis B, 9 with alcoholic liver disease, 14 with autoimmune hepatitis, 10 with primary biliary cirrhosis, and 5 of normal liver) were enrolled. The expression of the following genes was quantified by competitive reverse transcription-polymerase chain reaction and was compared according to the etiology; dsRNA-dependent protein kinase (PKR), 2',5' oligoadenylate synthetase (2,5-AS), latent cellular endoribonuclease (RNase L), RNase L inhibitor, and MxA. As a result, PKR mRNA was significantly overexpressed in the liver of chronic hepatitis C compared with those of other etiologies (P =.0178), and it correlated significantly with serum alanine transaminase values (r =.51, P =.0054). Also, the expression of the RNase L inhibitor showed a significant reduction in chronic hepatitis C (P =.0184). The expressions of 2,5 AS, RNase L, and MxA were not different significantly irrespective to the etiology. In conclusion, hepatic overexpression of PKR and reduced expression of RNase L inhibitor seem to contribute to the anti-HCV mechanism characteristically. PMID- 11050061 TI - Quantitative DNA fragment analysis for detecting low amounts of hepatitis B virus deletion mutants in highly viremic carriers. AB - Many variants of hepatitis B virus (HBV) with deletions in the viral genome have been identified. Some of these variants are indicator or even effector of a more severe course of hepatitis. These deletion mutants contribute a variable and sometimes very low proportion to the viral population. For early detection of small amounts of deletion mutants among a large number of wild-type genomes, we applied a new screening method designated quantitative fragment analysis (QFA). By QFA the whole viral genome can be scanned for the presence of deletions or insertions of >/=3 nucleotides representing more than 2% of the viral population. Using QFA we showed that an often described deletion of 8 nucleotides is packaged in viral capsids and not a polymerase chain reaction (PCR) artifact. QFA was applied to study the emergence of deletion mutants in a group of 18 pediatric patients who had been infected from a common source while being under multidrug cancer chemotherapy. All patients had developed a highly viremic asymptomatic HBV carrier state. In 3 of these patients 3 different kinds of HBV deletion mutants were found by QFA: 8 bp deletions within the core promoter, core gene deletions from 8 to 86 bp, and large deletions of up to 1,989 bp spanning the precore/core and the preS/S reading frames. PCR primers that specifically amplify deletion variants enabled the detection of additional patients harboring the investigated variant. PMID- 11050062 TI - Virologic and clinical expressions of reciprocal inhibitory effect of hepatitis B, C, and delta viruses in patients with chronic hepatitis. AB - We studied 648 hepatitis B surface antigen (HBsAg)- and/or anti-hepatitis C virus (HCV)-positive patients to evaluate the virologic and clinical characteristics of multiple hepatitis viral infection. We defined as Case B-C an HBsAg/anti-HCV positive patient and as Case b-C an anti-HCV/anti-HBc-positive, HBsAg/anti-HBs negative patient. For each Case B-C we scheduled as Control-B an HBsAg positive and anti-HCV negative patient and as Control-C an HBsAg/anti-HBs/anti-hepatitis B core antigen (HBc)-negative and anti-HCV-positive patient. Control group C was used as the control also for Case group b-C. Serum HBV DNA by molecular hybridization was found more frequently in Control group B (54% of 161 patients) than in Case group B-C (35.7% of 84, P <.01). The prevalence of HBV wild type was similar in Case group B-C (14. 3%) and in Control group B (17.4%), whereas the e minus strain was less frequent in Case group B-C (10.7% vs. 33%; P <.01). HBV DNA by polymerase chain reaction (PCR) was detected in 40.8% of 71 patients in Case group b-C. HCV RNA was detected more frequently in Control group C (90.7% of 130 patients) than in Case group B-C (65.2% of 69, P <.0001). Moderate or severe chronic hepatitis or cirrhosis were more frequent in Case group B-C (62.9% of 65 patients) than in Control group B (46.7% of 90, P <.05) or C (40.8% of 98, P <.005), and in Case group b-C (71.1% of 76) than in Control group C. Thus, in multiple hepatitis we observed a reciprocal inhibition of the viral genomes and a more severe liver disease. In Case group b-C, serum HBV DNA was frequent and the clinical presentation was severe. PMID- 11050063 TI - Immunohistochemical differentiation of hepatitis D virus genotypes. AB - Determination of hepatitis D virus (HDV) genotypes is epidemiologically and clinically important. Phylogenic analysis based on sequencing analysis of multiple HDV strains isolated from sera of patients is not convenient for mass screening in routine laboratories. This study was designed to develop genotype specific antibodies against hepatitis delta antigen (HDAg) and to apply these antibodies for immunohistochemical differentiation of HDV genotypes in formalin fixed, paraffin-embedded liver biopsies of patients. Divergence in the carboxyl terminal 19 amino acids of the large HDAg between genotypes I and II is more than 70%. Peptides covering these residues were conjugated to keyhole limpet hemocyanin and were used for immunization. The generated antibodies were confirmed for their specificity by binding to type-specific HDAgs expressed in DNA-transfected Huh-7 hepatoma cells. Liver biopsies from 6 patients who had dominant genotype I HDV and 33 patients who had dominant genotype II HDV in sera were stained with these antibodies. The accuracy for these antibodies was 94.9%, and the agreement between dominant HDV genotypes in serum and dominant hepatic HDV genotypes based on HDAg staining was nearly perfect (kappa = 0.83). In summary, the carboxyl-terminal 19 amino acids of the large HDAg can be used as immunogens to generate genotype-specific antibodies. These antibodies were proven to be useful in immunohistochemical differentiation of HDV genotypes in liver biopsies. PMID- 11050064 TI - Incubation phase of acute hepatitis B in man: dynamic of cellular immune mechanisms. AB - After hepatitis B virus (HBV) infection, liver injury and viral control have been thought to result from lysis of infected hepatocytes by virus-specific cytotoxic T cells. Patients are usually studied only after developing significant liver injury, and so the viral and immune events during the incubation phase of disease have not been defined. During a single-source outbreak of HBV infection, we identified patients before the onset of symptomatic hepatitis. The dynamics of HBV replication, liver injury, and HBV-specific CD8+ and CD4+ cell responses were investigated from incubation to recovery. Although a rise in alanine transaminase (ALT) levels was present at the time of the initial fall in HBV-DNA levels, maximal reduction in virus level occurred before significant liver injury. Direct ex vivo quantification of HBV-specific CD4+ and CD8+ cells, by using human leukocyte antigen (HLA) class I tetramers and intracellular cytokine staining, showed that adaptive immune mechanisms are present during the incubation phase, at least 4 weeks before symptoms. The results suggest that the pattern of reduction in HBV replication is not directly proportional to tissue injury during acute hepatitis B in humans. Furthermore, because virus-specific immune responses and significant reductions in viral replication are seen during the incubation phase, it is likely that the immune events central to viral control occur before symptomatic disease. PMID- 11050065 TI - Early identification of recipients with progressive histologic recurrence of hepatitis C after liver transplantation. AB - Approximately half of patients undergoing liver transplantation (LT) for hepatitis C virus (HCV) develop histologic evidence of recurrence within the first postoperative year. Early identification of recipients at risk for more severe recurrence of HCV may be useful in selecting patients for antiviral therapy. We determined whether recipients at greatest risk for more severe recurrence of HCV can be identified by pre- and/or early post-LT HCV-RNA levels in serum or tissue. Serum and tissue samples were prospectively collected pre-LT and at 7 days, 4 months, 1 year, and at 3 years posttransplantation from patients undergoing LT for HCV. Hepatitis activity index (HAI) and fibrosis stage (FS) were assessed in all liver biopsies. Forty-seven patients (32 men) were studied. Higher HCV-RNA levels at 4 months post-LT (>/=10(9) copies/mL, n = 29) were associated with higher HAI at 1 year and at 3 years post-LT. The HAI seen on protocol biopsies at 4 months correlated significantly with fibrosis stage (FS) at 1 year (r =.56, P CA1 synapse of hippocampus by stimulation in the theta frequency range (5-12 Hz), an effect that depends on activation of the cAMP pathway. We investigated the mechanisms of the cAMP contribution to this form of LTP in the rat hippocampal slice preparation. theta pulse stimulation (TPS; 150 stimuli at 10 Hz) by itself did not induce LTP, but the addition of either the beta-adrenergic agonist isoproterenol or the cAMP analog 8-bromo-cAMP (8-Br-cAMP) enabled TPS-induced LTP. The isoproterenol effect was blocked by postsynaptic inhibition of cAMP dependent protein kinase. Several lines of evidence indicated that cAMP enabled LTP by blocking postsynaptic protein phosphatase-1 (PP1). Activators of the cAMP pathway reduced PP1 activity in the CA1 region and increased the active form of inhibitor-1, an endogenous inhibitor of PP1. Postsynaptic injection of activated inhibitor-1 mimicked the LTP-enabling effect of cAMP pathway stimulation. TPS evoked complex spiking when isoproterenol was present. However, complex spiking was not sufficient to enable TPS-induced LTP, which additionally required the inhibition of postsynaptic PP1. PP1 inhibition seems to promote the activation of Ca(2+)/calmodulin-dependent protein kinase (CaMKII), because (1) a CaMKII inhibitor blocked the induction of LTP by TPS paired with either isoproterenol or activated inhibitor-1 and (2) CaMKII in area CA1 was activated by the combination of TPS and 8-Br-cAMP but not by either stimulus alone. These results indicate that the cAMP pathway enables TPS-induced LTP by inhibiting PP1, thereby enhancing Ca(2+)-independent CaMKII activity. PMID- 11050108 TI - Reduction in the density and expression, but not G-protein coupling, of serotonin receptors (5-HT1A) in 5-HT transporter knock-out mice: gender and brain region differences. AB - The aim of the present study was to investigate the mechanisms underlying the desensitization of 5-HT(1A) receptors in the dorsal raphe and hypothalamus of serotonin (5-HT) transporter knock-out mice (5-HTT -/-). The density of 5-HT(1A) receptors in the dorsal raphe was reduced in both male and female 5-HTT -/- mice. This reduction was more extensive in female than in male 5-HTT -/- mice. 8-OH DPAT-induced hypothermia was absent in female 5-HTT -/- and markedly attenuated in 5-HTT +/- mice. The density of 5-HT(1A) receptors also was decreased significantly in several nuclei of the hypothalamus, amygdala, and septum of female 5-HTT -/- mice. 5-HT(1A) receptor mRNA was reduced significantly in the dorsal raphe region, but not in the hypothalamus or hippocampus, of female 5-HTT +/- and 5-HTT -/- mice. G-protein coupling to 5-HT(1A) receptors and G-protein levels in most brain regions were not reduced significantly, except that G(o) and G(i1) proteins were reduced modestly in the midbrain of 5-HTT -/- mice. These data suggest that the desensitization of 5-HT(1A) receptors in 5-HTT -/- mice may be attributable to a reduction in the density of 5-HT(1A) receptors. This reduction is brain region-specific and more extensive in the female mice. The reduction in the density of 5-HT(1A) receptors may be mediated partly by reduction in the gene expression of 5-HT(1A) receptors in the dorsal raphe, but also by other mechanisms in the hypothalamus of 5-HTT -/- female mice. Finally, alterations in G-protein coupling to 5-HT(1A) receptors are unlikely to be involved in the desensitization of 5-HT(1A) receptors in 5-HTT -/- mice. PMID- 11050109 TI - Selective blockade of P/Q-type calcium channels by the metabotropic glutamate receptor type 7 involves a phospholipase C pathway in neurons. AB - Although presynaptic localization of mGluR7 is well established, the mechanism by which the receptor may control Ca(2+) channels in neurons is still unknown. We show here that cultured cerebellar granule cells express native metabotropic glutamate receptor type 7 (mGluR7) in neuritic processes, whereas transfected mGluR7 was also expressed in cell bodies. This allowed us to study the effect of the transfected receptor on somatic Ca(2+) channels. In transfected neurons, mGuR7 selectively inhibited P/Q-type Ca(2+) channels. The effect was mimicked by GTPgammaS and blocked by pertussis toxin (PTX) or a selective antibody raised against the G-protein alphao subunit, indicating the involvement of a G(o)-like protein. The mGuR7 effect did not display the characteristics of a direct interaction between G-protein betagamma subunits and the alpha1A Ca(2+) channel subunit, but was abolished by quenching betagamma subunits with specific intracellular peptides. Intracellular dialysis of G-protein betagamma subunits did not mimic the action of mGluR7, suggesting that both G-protein betagamma and alphao subunits were required to mediate the effect. Inhibition of phospholipase C (PLC) blocked the inhibitory action of mGluR7, suggesting that a coincident activation of PLC by the G-protein betagamma with alphao subunits was required. The Ca(2+) chelator BAPTA, as well as inhibition of either the inositol trisphosphate (IP(3)) receptor or protein kinase C (PKC) abolished the mGluR7 effect. Moreover, activation of native mGluR7 induced a PTX-dependent IP(3) formation. These results indicated that IP(3)-mediated intracellular Ca(2+) release was required for PKC-dependent inhibition of the Ca(2+) channels. Possible control of synaptic transmission by the present mechanisms is discussed. PMID- 11050110 TI - Role of bicarbonate and chloride in GABA- and glycine-induced depolarization and [Ca2+]i rise in fetal rat motoneurons in situ. AB - Ca(2+) imaging and (perforated) patch recording were used to analyze the mechanism of GABA- and glycine-induced depolarizations in lumbar motoneurons of spinal cord slices from fetal rats. In fura-2 ester-loaded cells, the agonist induced depolarizations increased [Ca(2+)](i) by up to 100 nm. The GABA- and glycine-evoked [Ca(2+)](i) transients were suppressed by bicuculline and strychnine, respectively. Their magnitude decreased by approximately 50% between embryonic days 15.5 and 19.5. The [Ca(2+)](i) increases were abolished by Ca(2+) free superfusate and attenuated by approximately 65% by nifedipine, showing that the responses were mediated by voltage-activated Ca(2+) channels. The [Ca(2+)](i) rises were potentiated by >300% immediately after removal of Cl(-) from the superfusate but recovered to values of 50-200% of control during repeated agonist administration in Cl(-)-free saline. Bumetanide gradually suppressed the [Ca(2+)](i) increases by >75%. Subsequent removal of Cl(-) reconstituted the responses and increased, upon repeated agonist application, the peak [Ca(2+)](i) rises to values above control. Removal of HCO(3)(-) from the Cl(-)-free (bumetanide-containing) superfusate reversibly abolished both the agonist-induced [Ca(2+)](i) rises and depolarizations that were reestablished by formate anions. In Cl(-)-containing superfusate, removal of HCO(3)(-) decreased both the peak and duration of the agonist-evoked membrane depolarization and [Ca(2+)](i) response. Our findings show that HCO(3)(-) efflux has a major contribution to depolarizations mediated by GABA(A) and glycine receptor-coupled anion channels in prenatal neurons. We hypothesize that the HCO(3)(-)-dependent depolarizing component, which is likely to produce an intracellular acidosis, might play an important role during the early postnatal period when the Cl(-)-dependent component gradually shifts to hyperpolarization. PMID- 11050111 TI - Slow desensitization regulates the availability of synaptic GABA(A) receptors. AB - At central synapses, a large and fast spike of neurotransmitter efficiently activates postsynaptic receptors. However, low concentrations of transmitter can escape the cleft and activate presynaptic and postsynaptic receptors. We report here that low concentrations of GABA reduce IPSCs in hippocampal neurons by preferentially desensitizing rather than opening GABA(A) channels. GABA transporter blockade also caused desensitization by locally elevating GABA to approximately 1 microm. Recovery of the IPSC required several seconds, mimicking recovery of the channel from slow desensitization. These results indicate that low levels of GABA can regulate the amplitude of IPSCs by producing a slow form of receptor desensitization. Accumulation of channels in this absorbing state allows GABA(A) receptors to detect even a few molecules of GABA in the synaptic cleft. PMID- 11050112 TI - Developmental changes in synaptic AMPA and NMDA receptor distribution and AMPA receptor subunit composition in living hippocampal neurons. AB - AMPA and NMDA receptors mediate most excitatory synaptic transmission in the CNS. We have developed antibodies that recognize all AMPA or all NMDA receptor variants on the surface of living neurons. AMPA receptor variants were identified with a polyclonal antibody recognizing the conserved extracellular loop region of all four AMPA receptor subunits (GluR1-4, both flip and flop), whereas NMDA receptors were immunolabeled with a polyclonal antibody that binds to an extracellular N-terminal epitope of the NR1 subunit, common to all splice variants. In non-fixed brain sections these antibodies gave labeling patterns similar to autoradiographic distributions with particularly high levels in the hippocampus. Using these antibodies, in conjunction with GluR2-specific and synaptophysin antibodies, we have directly localized and quantified surface expressed native AMPA and NMDA receptors on cultured living hippocampal neurons during development. Using a quantitative cell ELISA, a dramatic increase was observed in the surface expression of AMPA receptors, but not NMDA receptors, between 3 and 10 d in culture. Immunocytochemical analysis of hippocampal neurons between 3 and 20 d in vitro shows no change in the proportion of synapses expressing NMDA receptors (approximately 60%) but a dramatic increase (approximately 50%) in the proportion of them that also express AMPA receptors. Furthermore, over this period the proportion of AMPA receptor-positive synapses expressing the GluR2 subunit increased from approximately 67 to approximately 96%. These changes will dramatically alter the functional properties of hippocampal synapses. PMID- 11050113 TI - Regulation of AMPA receptor GluR1 subunit surface expression by a 4. 1N-linked actin cytoskeletal association. AB - The synaptic localization, clustering, and immobilization of neurotransmitter receptors and ion channels play important roles in synapse formation and synaptic transmission. Although several proteins have been identified that interact with AMPA receptors and that may regulate their synaptic targeting, little is known about the interaction of AMPA receptors with the cytoskeleton. In studies examining the interaction of the AMPA receptor GluR1 subunit with neuronal proteins, we determined that GluR1 interacts with the 4.1G and 4.1N proteins, homologs of the erythrocyte membrane cytoskeletal protein 4.1. Using the yeast two-hybrid system and a heterologous cell system, we demonstrated that both 4.1G and 4.1N bind to a membrane proximal region of the GluR1 C terminus, and that a region within the C-terminal domain of 4.1G or 4.1N is sufficient to mediate the interaction. We also found that 4.1N can associate with GluR1 in vivo and colocalizes with AMPA receptors at excitatory synapses. Disruption of the interaction of GluR1 with 4.1N or disruption of actin filaments decreased the surface expression of GluR1 in heterologous cells. Moreover, disruption of actin filaments in cultured cortical neurons dramatically reduced the level of surface AMPA receptors. These results suggest that protein 4.1N may link AMPA receptors to the actin cytoskeleton. PMID- 11050114 TI - Novel SCAMPs lacking NPF repeats: ubiquitous and synaptic vesicle-specific forms implicate SCAMPs in multiple membrane-trafficking functions. AB - In vertebrates, secretory carrier membrane proteins (SCAMPs) 1-3 constitute a family of putative membrane-trafficking proteins composed of cytoplasmic N terminal sequences with NPF repeats, four central transmembrane regions (TMRs), and a cytoplasmic tail. SCAMPs probably function in endocytosis by recruiting EH domain proteins to the N-terminal NPF repeats but may have additional functions mediated by their other sequences. We now demonstrate that SCAMPs form a much larger and more heterogeneous protein family than envisioned previously, with an evolutionary conservation extending to invertebrates and plants. Two novel vertebrate SCAMPs (SCAMPs 4 and 5), single SCAMP genes in Caenorhabditis elegans and Drosophila melanogaster, and multiple SCAMPs in Arabidopsis thaliana were identified. Interestingly, the novel SCAMPs 4 and 5 lack the N-terminal NPF repeats that are highly conserved in all other SCAMPs. RNA and Western blotting experiments showed that SCAMPs 1-4 are ubiquitously coexpressed, whereas SCAMP 5 is only detectable in brain where it is expressed late in development coincident with the elaboration of mature synapses. Immunocytochemistry revealed that SCAMP 5 exhibits a synaptic localization, and subcellular fractionations demonstrated that SCAMP 5 is highly enriched in synaptic vesicles. Our studies characterize SCAMPs as a heterogeneous family of putative trafficking proteins composed of three isoforms that are primarily synthesized outside of neurons (SCAMPs 2-4), one isoform that is ubiquitously expressed but highly concentrated on synaptic vesicles (SCAMP 1), and one brain-specific isoform primarily localized to synaptic vesicles (SCAMP 5). The conservation of the TMRs in all SCAMPs with the variable presence of N-terminal NPF repeats suggests that in addition to the role of some SCAMPs in endocytosis mediated by their NPF repeats, all SCAMPs perform a "core" function in membrane traffic mediated by their TMRs. PMID- 11050115 TI - Mice with combined gene knock-outs reveal essential and partially redundant functions of amyloid precursor protein family members. AB - The amyloid precursor protein (APP) involved in Alzheimer's disease is a member of a larger gene family including amyloid precursor-like proteins APLP1 and APLP2. We generated and examined the phenotypes of mice lacking individual or all possible combinations of APP family members to assess potential functional redundancies within the gene family. Mice deficient for the nervous system specific APLP1 protein showed a postnatal growth deficit as the only obvious abnormality. In contrast to this minor phenotype, APLP2(-/-)/APLP1(-/-) and APLP2(-/-)/APP(-/-) mice proved lethal early postnatally. Surprisingly, APLP1(-/ )/APP(-/-) mice were viable, apparently normal, and showed no compensatory upregulation of APLP2 expression. These data indicate redundancy between APLP2 and both other family members and corroborate a key physiological role for APLP2. This view gains further support by the observation that APLP1(-/-)/APP(-/ )/APLP2(+/-) mice display postnatal lethality. In addition, they provide genetic evidence for at least some distinct physiological roles of APP and APLP2 by demonstrating that combinations of single knock-outs with the APLP1 mutation resulted in double mutants of clearly different phenotypes, being either lethal, or viable. None of the lethal double mutants displayed, however, obvious histopathological abnormalities in the brain or any other organ examined. Moreover, cortical neurons from single or combined mutant mice showed unaltered survival rates under basal culture conditions and unaltered susceptibility to glutamate excitotoxicity in vitro. PMID- 11050116 TI - Tumor necrosis factor alpha induces a metalloprotease-disintegrin, ADAM8 (CD 156): implications for neuron-glia interactions during neurodegeneration. AB - ADAM proteases, defined by extracellular disintegrin and metalloprotease domains, are involved in protein processing and cell-cell interactions. Using wobbler (WR) mutant mice, we investigated the role of ADAMs in neurodegeneration and reactive glia activation in the CNS. We found that ADAM8 (CD 156), a suspected leukocyte adhesion molecule, is expressed in the CNS and highly induced in affected CNS areas of WR mice, in brainstem and spinal cord. ADAM8 mRNA and protein are found at low levels throughout the normal mouse CNS, in neurons and oligodendrocytes. In the WR CNS regions in which neurodegeneration occurs, ADAM8 is induced in neurons, reactive astrocytes, and activated microglia. Similarly, the proinflammatory cytokine tumor necrosis factor alpha (TNF-alpha) is upregulated and shows the same cellular distribution. In primary astrocytes from wild-type and WR mice, in primary cerebellar neurons, and in mouse motoneuron-like NSC19 cells, ADAM8 expression was induced up to 15-fold by mouse TNF-alpha, in a dose dependent manner. In both cell types, ADAM8 was also induced by human TNF-alpha, indicating that TNF receptor type I (p55) is involved. Induction of ADAM8 mRNA was suppressed by treatment with an interferon-regulating factor 1 (IRF-1) antisense oligonucleotide. We conclude that IRF-1-mediated induction of ADAM8 by TNF-alpha is a signaling pathway relevant for neurodegenerative disorders with glia activation, proposing a role for ADAM8 in cell adhesion during neurodegeneration. PMID- 11050117 TI - Constitutive endocytosis of GABAA receptors by an association with the adaptin AP2 complex modulates inhibitory synaptic currents in hippocampal neurons. AB - Type A GABA receptors (GABA(A)) mediate the majority of fast synaptic inhibition in the brain and are believed to be predominantly composed of alpha, beta, and gamma subunits. Although changes in cell surface GABA(A) receptor number have been postulated to be of importance in modulating inhibitory synaptic transmission, little is currently known on the mechanism used by neurons to modify surface receptor levels at inhibitory synapses. To address this issue, we have studied the cell surface expression and maintenance of GABA(A) receptors. Here we show that constitutive internalization of GABA(A) receptors in hippocampal neurons and recombinant receptors expressed in A293 cells is mediated by clathrin-dependent endocytosis. Furthermore, we identify an interaction between the GABA(A) receptor beta and gamma subunits with the adaptin complex AP2, which is critical for the recruitment of integral membrane proteins into clathrin-coated pits. GABA(A) receptors also colocalize with AP2 in cultured hippocampal neurons. Finally, blocking clathrin-dependant endocytosis with a peptide that disrupts the association between amphiphysin and dynamin causes a large sustained increase in the amplitude of miniature IPSCs in cultured hippocampal neurons. These results suggest that GABA(A) receptors cycle between the synaptic membrane and intracellular sites, and their association with AP2 followed by recruitment into clathrin-coated pits represents an important mechanism in the postsynaptic modulation of inhibitory synaptic transmission. PMID- 11050118 TI - In situ Ca2+ imaging reveals neurotransmitter receptors for glutamate in taste receptor cells. AB - The neurotransmitters at synapses in taste buds are not yet known with confidence. Here we report a new calcium-imaging technique for taste buds that allowed us to test for the presence of glutamate receptors (GluRs) in living isolated tissue preparations. Taste cells of rat foliate papillae were loaded with calcium green dextran (CaGD). Lingual slices containing CaGD-labeled taste cells were imaged with a scanning confocal microscope and superfused with glutamate (30 micromter to 1 mm), kainate (30 and 100 micrometer), AMPA (30 and 100 micrometer), or NMDA (100 micrometer). Responses were observed in 26% of the cells that were tested with 300 micrometer glutamate. Responses to glutamate were localized to the basal processes and cell bodies, which are synaptic regions of taste cells. Glutamate responses were dose-dependent and were induced by concentrations as low as 30 microm. The non-NMDA receptor antagonists CNQX and GYKI 52466 reversibly blocked responses to glutamate. Kainate, but not AMPA, also elicited Ca(2+) responses. NMDA stimulated increases in [Ca(2+)](i) when the bath medium was modified to optimize for NMDA receptor activation. The subset of cells that responded to glutamate was either NMDA-unresponsive (54%) or NMDA-responsive (46%), suggesting that there are presumably two populations of glutamate sensitive taste cells-one with NMDA receptors and the other without NMDA receptors. The function of GluRs in taste buds is not yet known, but the data suggest that glutamate is a neurotransmitter there. GluRs in taste cells might be presynaptic autoreceptors or postsynaptic receptors at afferent or efferent synapses. PMID- 11050119 TI - Clathrin-mediated endocytosis near active zones in snake motor boutons. AB - We have used the activity-dependent probe FM1-43 with electron microscopy (EM) to examine endocytosis at the vertebrate nerve-muscle synapse. Preparations were fixed after very brief neural stimulation at reduced temperature, and internalized FM1-43 was photoconverted into an electron-dense reaction product. To locate the reaction product, we reconstructed computer renderings of individual terminal boutons from serial EM sections. Most of the reaction product was seen in 40-60 nm vesicles. All of the labeled vesicles were clathrin-coated, and 92% of them were located within 300 nm of the plasma membrane, suggesting that they had undergone little processing after retrieval from their endocytic sites. The vesicles (and by inference the sites) were not dispersed randomly near the plane of the membrane but instead were clustered significantly near active zones. Additional reaction product was found within putative macropinosomes; these appeared to form from deep membrane invaginations near active zones. Thus two mechanisms of endocytosis were evident after brief stimulation. Endocytosis near active zones is consistent with the existence of local exo/endocytic cycling pools. This mechanism also might serve to maintain alignment of active zones with postsynaptic folds during periods of activity when vesicular and plasma membranes are interchanged. PMID- 11050120 TI - Apoptosis has a prolonged role in the neurodegeneration after hypoxic ischemia in the newborn rat. AB - Birth asphyxia can cause moderate to severe brain injury. It is unclear to what degree apoptotic or necrotic mechanisms of cell death account for damage after neonatal hypoxia-ischemia (HI). In a 7-d-old rat HI model, we determined the contributions of apoptosis and necrosis to neuronal injury in adjacent Nissl stained, hematoxylin and eosin-stained, and terminal deoxynucleotidyl transferase mediated UTP nick end-labeled sections. We found an apoptotic-necrotic continuum in the morphology of injured neurons in all regions examined. Eosinophilic necrotic neurons, typical in adult models, were rarely observed in neonatal HI. Electron microscopic analysis showed "classic" apoptotic and necrotic neurons and "hybrid" cells with intermediate characteristics. The time course of apoptotic injury varied regionally. In CA3, dentate gyrus, medial habenula, and laterodorsal thalamus, the density of apoptotic cells was highest at 24-72 hr after HI and then declined. In contrast, densities remained elevated from 12 hr to 7 d after HI in most cortical areas and in the basal ganglia. Temporal and regional patterns of neuronal death were compared with expression of caspase-3, a cysteine protease involved in the execution phase of apoptosis. Immunocytochemical and Western blot analyses showed increased caspase-3 expression in damaged hemispheres 24 hr to 7 d after HI. A p17 peptide fragment, which results from the proteolytic activation of the caspase-3 precursor, was detected in hippocampus, thalamus, and striatum but not in cerebral cortex. The continued expression of activated caspase-3 and the persistence of cells with an apoptotic morphology for days after HI suggests a prolonged role for apoptosis in neonatal hypoxic ischemic brain injury. PMID- 11050121 TI - NMDA but not non-NMDA excitotoxicity is mediated by Poly(ADP-ribose) polymerase. AB - Poly(ADP-ribose) polymerase (PARP-1), a nuclear enzyme that facilitates DNA repair, may be instrumental in acute neuronal cell death in a variety of insults including, cerebral ischemia, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced parkinsonism, and CNS trauma. Excitotoxicity is thought to underlie these and other toxic models of neuronal death. Different glutamate agonists may trigger different downstream pathways toward neurotoxicity. We examine the role of PARP-1 in NMDA- and non-NMDA-mediated excitotoxicity. NMDA and non-NMDA agonists were stereotactically delivered into the striatum of mice lacking PARP-1 and control mice in acute (48 hr) and chronic (3 week) toxicity paradigms. Mice lacking PARP-1 are highly resistant to the excitoxicity induced by NMDA but are as equally susceptible to AMPA excitotoxicity as wild-type mice. Restoring PARP-1 protein in mice lacking PARP-1 by viral transfection restored susceptibility to NMDA, supporting the requirement of PARP-1 in NMDA neurotoxicity. Furthermore, Western blot analyses demonstrate that PARP-1 is activated after NMDA delivery but not after AMPA administration. Consistent with the theory that nitric oxide (NO) and peroxynitrite are prominent in NMDA-induced neurotoxicity, PARP-1 was not activated in mice lacking the gene for neuronal NO synthase after NMDA administration. These results suggest a selective role of PARP-1 in glutamate excitoxicity, and strategies of inhibiting PARP-1 in NMDA-mediated neurotoxicity may offer substantial acute and chronic neuroprotection. PMID- 11050123 TI - Cell migration and aggregation in the developing telencephalon: pulse-labeling chick embryos with bromodeoxyuridine. AB - Previous studies had concluded that the avian telencephalon develops according to an outside-in schedule of neurogenesis, with relatively little migration of young neuroblasts past older cells. These previous studies had, however, been based on the "cumulative labeling" method, which is less accurate than the "pulse labeling" method typically used in mammals. In the present study, we pulse labeled chick embryos by injecting low doses of the thymidine analog bromodeoxyuridine (BrdU) directly into the circulatory system of chick embryos at 6 d of incubation. The brains of these embryos were then examined for anti-BrdU labeled cells at postinjection survival times from 30 min to 10 d. Comparisons across different survival times, as well as with cases in which BrdU was injected on day 7, suggested that our effective pulse duration is <24 hr. This was confirmed by injecting tritiated thymidine 24 hr after the BrdU and seeing no double-labeled cells. Several deviations from the previously reported pattern of telencephalic neurogenesis were also noted. Most importantly, the cells born on day 6 in the avian Wulst, the likely homolog of mammalian neocortex, end up homogeneously distributed throughout the Wulst, which suggests that many of them are migrating past older cells. Furthermore, the cells born on day 6 in the ventral hyperstriatum and dorsal neostriatum gradually (over the course of 2-3 d) aggregate into distinct multicellular clusters, which suggests that isochronic cells in these regions adhere preferentially to one another. Finally, the data reveal a proliferative subventricular zone similar to that observed in the ganglionic eminences of mammalian embryos. PMID- 11050122 TI - A glia-derived signal regulating neuronal differentiation. AB - Astrocytes are present in large numbers in the nervous system, are associated with synapses, and propagate ionic signals. Astrocytes influence neuronal physiology by responding to and releasing neurotransmitters, but the mechanisms that establish the close interaction between these cells are not defined. Here we use hippocampal neurons in culture to demonstrate that vasoactive intestinal polypeptide (VIP) promotes neuronal differentiation through activity-dependent neurotrophic factor (ADNF), a protein secreted by VIP-stimulated astroglia. ADNF is produced by glial cells and acts directly on neurons to promote glutamate responses and morphological development. ADNF causes secretion of neurotrophin 3 (NT-3), and both proteins regulate NMDA receptor subunit 2A (NR2A) and NR2B. These data suggest that the VIP-ADNF-NT-3 neuronal-glial pathway regulates glutamate responses from an early stage in the synaptic development of excitatory neurons and may also contribute to the known effects of VIP on learning and behavior in the adult nervous system. PMID- 11050124 TI - A purine-sensitive pathway regulates multiple genes involved in axon regeneration in goldfish retinal ganglion cells. AB - In lower vertebrates, retinal ganglion cells (RGCs) can regenerate their axons and reestablish functional connections after optic nerve injury. We show here that in goldfish RGCs, the effects of several trophic factors converge on a purine-sensitive signaling mechanism that controls axonal outgrowth and the expression of multiple growth-associated proteins. In culture, goldfish RGCs regenerate their axons in response to two molecules secreted by optic nerve glia, axogenesis factor-1 (AF-1) and AF-2, along with ciliary neurotrophic factor. The purine analog 6-thioguanine (6-TG) blocked outgrowth induced by each of these factors. Previous studies in PC12 cells have shown that the effects of 6-TG on neurite outgrowth may be mediated via inhibition of a 47 kDa protein kinase. Growth factor-induced axogenesis in RGCs was accompanied by many of the molecular changes that characterize regenerative growth in vivo, e.g. , increased expression of GAP-43 and certain cell surface glycoproteins. 6-TG inhibited all of these changes but not those associated with axotomy per se, e.g., induction of jun family transcription factors, nor did it affect cell survival. Additional studies using RGCs from transgenic zebrafish showed that expression of Talpha-1 tubulin is likewise stimulated by AF-1 and blocked by 6-TG. The purine nucleoside inosine had effects opposite to those of 6-TG. Inosine stimulated outgrowth and the characteristic pattern of molecular changes in RGCs and competitively reversed the inhibitory effects of 6-TG. We conclude that axon regeneration and the underlying program of gene expression in goldfish RGCs are mediated via a common, purine-sensitive pathway. PMID- 11050125 TI - Pax6 modulates the dorsoventral patterning of the mammalian telencephalon. AB - The Pax6 gene encodes a transcription factor with a restricted expression in the ventricular zone of the pallium and subpallium. We tested whether the function of Pax6 is necessary for the correct patterning and morphogenesis of the vertebrate telencephalon. Homozygous embryos of the Pax6/Small eye mutant lack functional PAX6 protein because of a point mutation of the gene. In the mutant Small eye embryos we detected a ventralization of the molecular patterning of the telencephalon at two borders, the pallium/subpallium and the lateral/medial ganglionic eminence. The results indicate that Pax6 controls the lateral limit of the expression of Nkx2.1, Shh, and Lhx6 in the prechordal neural tube, the telencephalon. This finding is in agreement with previous studies and supports a model for a common genetic mechanism for modulation of the dorsoventral patterning of the prechordal and epichordal CNS. The pattern defects caused by the loss of Pax6 function result in multiple morphological abnormalities in the Small eye brain: dysgenesis of the piriform, insular, and lateral cortices, the claustrum-endopiriform nucleus, and a failure in the differentiation of a subpopulation of the cortical precursors. Together the results demonstrate that Pax6 has an essential role for the modulation of the dorsoventral patterning of the embryonic telencephalon, influencing thereby the forebrain morphogenesis. PMID- 11050126 TI - Activity-dependent patterning of retinogeniculate axons proceeds with a constant contribution from AMPA and NMDA receptors. AB - Neural activity is critical for the refinement of neural circuitry during development, although the mechanisms involved in stabilizing appropriate connectivity remain unclear. It has been proposed that the insertion of AMPA receptors at synapses with only NMDA receptors ("silent synapses") mediates this stabilization, leading to an increasing contribution from AMPA receptors as development proceeds. Here we show in a mammalian system known to undergo activity-dependent development [the segregation of retinal afferents into ON/OFF sublaminae in the ferret lateral geniculate nucleus (LGN)] that the refinement of the neural circuitry occurs in the presence of a constant functional contribution from AMPA and NMDA receptors. Although we detected a small number of silent synapses on LGN cells, their proportion did not decrease with age. The size and kinetics of NMDA-mediated spontaneous EPSCs (sEPSCs) were also stable over this period. Together with previous results reporting the stability of unitary AMPA mediated EPSCs, the constancy of NMDA-mediated sEPSCs indicates an unchanging AMPA/NMDA contribution. Additionally, the CNQX-sensitivity did not increase for either sEPSCs or optic tract-evoked EPSCs. Likewise, the anatomical AMPA/NMDA ratio, as assayed by quantifying the colocalized expression of AMPA and NMDA receptor subunits, was fixed throughout ON/OFF sublamination. In particular, the colocalization of AMPA receptor subunits (GluR1 or GluR4) and NMDA receptor subunit NR1 opposite identified retinogeniculate terminals was stable during this period. These results add to the view of the population of retinogeniculate synapses as robustly stable or normalized during a period when retinogeniculate axons are undergoing dramatic activity-dependent refinement. PMID- 11050127 TI - Regeneration of lesioned corticospinal tract fibers in the adult rat induced by a recombinant, humanized IN-1 antibody fragment. AB - Axons in the CNS of higher vertebrates generally fail to regenerate after injury. This lack of regeneration is crucially influenced by neurite growth inhibitory protein constituents of CNS myelin. We have shown previously that a monoclonal antibody (mAb IN-1) capable of binding and neutralizing Nogo-A, a myelin associated inhibitor of neurite growth, can induce long-distance axonal regeneration and increased structural plasticity with improved functional recovery in rat models of CNS injury. In this paper we demonstrate that a partially humanized, recombinant Fab fragment (rIN-1 Fab) derived from the original mAb IN-1, was able to promote long-distance regeneration of injured axons in the spinal cord of adult rats. When infused into a spinal cord injury site, regrowth of corticospinal fibers in 11 of 18 animals was observed after a survival time of 2 weeks. Regenerating fibers grew for >9 mm beyond the lesion site and arborized profusely in the distal cord. Regenerated fibers formed terminal arbors with varicosities in the spinal cord gray matter, strongly resembling synaptic points of contact to neurons in the spinal cord distal to the lesion. In animals that had received a bovine serum albumin solution or a recombinant IN-1 fragment that had been mutated in the antigen binding site (mutIN-1 Fab), no significant growth beyond normal lesion-induced sprouting was observed. Neutralization of endogenous nerve growth inhibitors represents a novel use of recombinant antibody technology with potential therapeutic applications after traumatic CNS lesions. PMID- 11050128 TI - Potassium-coupled chloride cotransport controls intracellular chloride in rat neocortical pyramidal neurons. AB - Chloride (Cl(-)) homeostasis is critical for many cell functions including cell signaling and volume regulation. The action of GABA at GABA(A) receptors is primarily determined by the concentration of intracellular Cl(-). Developmental regulation of intracellular Cl(-) results in a depolarizing response to GABA in immature neocortical neurons and a hyperpolarizing or shunting response in mature neocortical neurons. One protein that participates in Cl(-) homeostasis is the neuron-specific K(+)-Cl(-) cotransporter (KCC2). Thermodynamic considerations predict that in the physiological ranges of intracellular Cl(-) and extracellular K(+) concentrations, KCC2 can act to either extrude or accumulate Cl(-). To test this hypothesis, we examined KCC2 function in pyramidal cells from rat neocortical slices in mature (18-28 d postnatal) and immature (3-6 d postnatal) rats. Intracellular Cl(-) concentration was estimated from the reversal potential of whole-cell currents evoked by local application of exogenous GABA. Both increasing and decreasing the extracellular K(+) concentration resulted in a concomitant change in intracellular Cl(-) concentration in neurons from mature rats. KCC2 inhibition by furosemide caused a change in the intracellular Cl(-) concentration that depended on the concentration of pipette Cl(-); in recordings with low pipette Cl(-), furosemide lowered intracellular Cl(-), whereas in recordings with elevated pipette Cl(-), furosemide raised intracellular Cl(-). In neurons from neonatal rats, manipulation of extracellular K(+) had no effect on intracellular Cl(-) concentration, consistent with the minimal KCC2 mRNA levels observed in neocortical neurons from immature animals. These data demonstrate a physiologically relevant and developmentally regulated role for KCC2 in Cl(-) homeostasis via both Cl(-) extrusion and accumulation. PMID- 11050129 TI - Transmitter-receptor interactions between growth cones of identified Lymnaea neurons determine target cell selection in vitro. AB - In addition to their involvement in transsynaptic communication in the adult nervous system, neurotransmitters also participate in many developmental events, such as neurite initiation and outgrowth. Although growth cones can release transmitters and are themselves sensitive to exogenously applied neurotransmitters, a direct causal relationship between the release of transmitter from one growth cone and its effect on another has not yet been demonstrated. In this study, we provide evidence that dopamine release from the growth cones of an identified Lymnaea neuron, right pedal dorsal 1 (RPeD1), differentially regulates the growth cone behavior of its in vivo target and nontarget neurons in vitro. In coculture, RPeD1 growth cones enhanced the rate of growth cone advance from target cells and synaptic connections developed immediately after contact. In contrast, RPeD1 growth cones not only inhibited the rate of growth cone advance from nontarget cells but they also induced growth cone collapse. Using a "sniffer cell" approach, we demonstrated that both RPeD1 growth cones and somata released dopamine, which can be detected at a distance of several hundred micrometers. RPeD1 somata were used to demonstrate that spontaneous release of dopamine also acted as a chemoattractant for target growth cones but as a chemorepellent for nontarget growth cones. These effects were mimicked by exogenous dopamine application, and both RPeD1 growth cone and soma induced effects were also blocked in the presence of dopamine receptor antagonists. This study emphasizes the importance of transmitter-receptor interactions between growth cones in target cell selection. PMID- 11050131 TI - Parallel instabilities of long-term potentiation, place cells, and learning caused by decreased protein kinase A activity. AB - To further elucidate the links among synaptic plasticity, hippocampal place cells, and spatial memory, place cells were recorded from wild-type mice and transgenic "R(AB)" mice with reduced forebrain protein kinase A (PKA) activity after introduction into a novel environment. Place cells in both strains were similar during the first exposure and were equally stable for recording sessions separated by 1 hr. Place cell stability in wild-type mice was unchanged for sessions separated by 24 hr but was reduced in R(AB) mice over the longer interval. This stability pattern parallels both the reduced late-phase long-term potentiation in hippocampal slices from R(AB) mice and the amnesia for context fear conditioning seen in R(AB) mice 24 but not 1 hr after training. The similar time courses of synaptic, network, and behavioral instability suggest that the genetic reduction of PKA activity is responsible for the defects at each level and support the idea that hippocampal synaptic plasticity is important in spatial memory. PMID- 11050130 TI - Neuronal activity and brain-derived neurotrophic factor regulate the density of inhibitory synapses in organotypic slice cultures of postnatal hippocampus. AB - Hippocampal interneurons inhibit pyramidal neurons through the release of the neurotransmitter GABA. Given the importance of this inhibition for the proper functioning of the hippocampus, the development of inhibitory synapses must be tightly regulated. In this study, the possibility that neuronal activity and neurotrophins regulate the density of GABAergic inhibitory synapses was investigated in organotypic slice cultures taken from postnatal day 7 rats. In hippocampal slices cultured for 13 d in the presence of the GABA(A) receptor antagonist bicuculline, the density of glutamic acid decarboxylase (GAD) 65 immunoreactive terminals was increased in the CA1 area when compared with control slices. Treatment with the glutamate receptor antagonist 6,7-dinitroquinoxaline 2,3-dione decreased the density of GAD65-immunoreactive terminals in the stratum oriens of CA1. These treatments had parallel effects on the density of GABA immunoreactive processes. Electron microscopic analysis after postembedding immunogold labeling with antibodies against GABA indicated that bicuculline treatment increased the density of inhibitory but not excitatory synapses. Application of exogenous BDNF partly mimicked the stimulatory effect of bicuculline on GAD65-immunoreactive terminals. Finally, antibodies against BDNF, but not antibodies against nerve growth factor, decrease the density of GAD65 immunoreactive terminals in bicuculline-treated slices. Thus, neuronal activity regulates the density of inhibitory synapses made by postnatal hippocampal interneurons, and BDNF could mediate part of this regulation. This regulation of the density of inhibitory synapses could represent a feedback mechanism aimed at maintaining an appropriate level of activity in the developing hippocampal networks. PMID- 11050132 TI - Muscarinic tone sustains impulse flow in the septohippocampal GABA but not cholinergic pathway: implications for learning and memory. AB - Systemic infusions of the muscarinic cholinergic receptor antagonists atropine and scopolamine (atr/scop) produce an amnesic syndrome in humans, subhuman primates, and rodents. In humans, this syndrome may resemble early symptoms of Alzheimer's disease. Behavioral studies in rats have demonstrated that the medial septum/diagonal band of Broca (MSDB), which sends cholinergic and GABAergic projections to the hippocampus, is a critical locus in mediating the amnesic effects of atr/scop. The amnesic effects of atr/scop in the MSDB have been presumed but not proven to be caused by a decrease in hippocampal acetylcholine (ACh) release after blockade of a muscarinic tone in the MSDB. Using electrophysiological recordings and fluorescent-labeling techniques to identify living septohippocampal neurons in rat brain slices, we now report that, contrary to current belief, a blockade of the muscarinic tone in the MSDB does not decrease impulse flow in the septohippocampal cholinergic pathway; instead, it decreases impulse flow in the septohippocampal GABAergic pathway via M(3) muscarinic receptors. We also report that the muscarinic tone in the MSDB is maintained by ACh that is released locally, presumably via axon collaterals of septohippocampal cholinergic neurons. As such, cognitive deficits that occur in various neurodegenerative disorders that are associated with a loss or atrophy of septohippocampal cholinergic neurons cannot be attributed solely to a decrease in hippocampal acetylcholine release. An additional, possibly more important mechanism may be the concomitant decrease in septohippocampal GABA release and a subsequent disruption in disinhibitory mechanisms in the hippocampus. Restoration of impulse flow in the septohippocampal GABA pathway, possibly via M(3) receptor agonists, may, therefore, be critical for successful treatment of cognitive deficits associated with neurodegenerative disorders such as Alzheimer's and Parkinson's disease. PMID- 11050133 TI - Long-term optical imaging and spectroscopy reveal mechanisms underlying the intrinsic signal and stability of cortical maps in V1 of behaving monkeys. AB - Explorations of learning and memory, other long-term plastic changes, and additional cognitive functions in the behaving primate brain would greatly benefit from the ability to image the functional architecture within the same patch of cortex, at the columnar level, for a long period of time. We developed methods for long-term optical imaging based on intrinsic signals and repeatedly visualized the same functional domains in behaving macaque cortex for a period extending over 1 year. Using optical imaging and imaging spectroscopy, we first explored the relationship between electrical activity and hemodynamic events in the awake behaving primate and compared it with anesthetized preparations. We found that, whereas the amplitude of the intrinsic signal was much larger in the awake animal, its temporal pattern was similar to that observed in the anesthetized animals. In both groups, deoxyhemoglobin concentration reached a peak 2-3 sec after stimulus onset. Furthermore, the early activity-dependent increase in deoxyhemoglobin concentration (the "initial dip") was far more tightly colocalized with electrical activity than the delayed increase in oxyhemoglobin concentration, known to be associated with an increase in blood flow. The implications of these results for improvement of the spatial resolution of blood oxygenation level-dependent functional magnetic resonance imaging are discussed. After the characterization of the intrinsic signal in the behaving primate, we used this new imaging method to explore the stability of cortical maps in the macaque primary visual cortex. Functional maps of orientation and ocular dominance columns were found to be stable for a period longer than 1 year. PMID- 11050134 TI - Intra-accumbens amphetamine increases the conditioned incentive salience of sucrose reward: enhancement of reward "wanting" without enhanced "liking" or response reinforcement. AB - Amphetamine microinjection into the nucleus accumbens shell enhanced the ability of a Pavlovian reward cue to trigger increased instrumental performance for sucrose reward in a pure conditioned incentive paradigm. Rats were first trained to press one of two levers to obtain sucrose pellets. They were separately conditioned to associate a Pavlovian cue (30 sec light) with free sucrose pellets. On test days, the rats received bilateral microinjection of intra accumbens vehicle or amphetamine (0.0, 2.0, 10.0, or 20.0 microgram/0.5 microliter), and lever pressing was tested in the absence of any reinforcement contingency, while the Pavlovian cue alone was freely presented at intervals throughout the session. Amphetamine microinjection selectively potentiated the cue-elicited increase in sucrose-associated lever pressing, although instrumental responding was not reinforced by either sucrose or the cue during the test. Intra accumbens amphetamine can therefore potentiate cue-triggered incentive motivation for reward in the absence of primary or secondary reinforcement. Using the taste reactivity measure of hedonic impact, it was shown that intra-accumbens amphetamine failed to increase positive hedonic reaction patterns elicited by sucrose (i.e., sucrose "liking") at doses that effectively increase sucrose "wanting." We conclude that nucleus accumbens dopamine specifically mediates the ability of reward cues to trigger "wanting" (incentive salience) for their associated rewards, independent of both hedonic impact and response reinforcement. PMID- 11050135 TI - Antagonism of the melanocortin system reduces cold and mechanical allodynia in mononeuropathic rats. AB - The presence of both pro-opiomelanocortin-derived peptides and melanocortin (MC) receptors in nociception-associated areas in the spinal cord suggests that, at the spinal level, the MC system might be involved in nociceptive transmission. In the present study, we demonstrate that a chronic constriction injury (CCI) to the rat sciatic nerve, a lesion that produces neuropathic pain, results in changes in the spinal cord MC system, as shown by an increased binding of (125)I-NDP-MSH to the dorsal horn. Furthermore, we investigated whether intrathecal administration (in the cisterna magna) of selective MC receptor ligands can affect the mechanical and cold allodynia associated with the CCI. Mechanical and cold allodynia were assessed by measuring withdrawal responses of the affected limb to von Frey filaments and withdrawal latencies upon immersion in a 4.5 degrees C water bath, respectively. We show that treatment with the MC receptor antagonist SHU9119 has a profound anti-allodynic effect, suggesting that the endogenous MC system has a tonic effect on nociception. In contrast, administration of the MC4 receptor agonists MTII and d-Tyr-MTII primarily increases the sensitivity to mechanical and cold stimulation. No antinociceptive action was observed after administration of the selective MC3 receptor agonist Nle-gamma-MSH. Together, our data suggest that the spinal cord MC system is involved in neuropathic pain and that the effects of MC receptor ligands on the responses to painful stimuli are exerted through the MC4 receptor. In conclusion, antagonism of the spinal melanocortin system might provide a new approach in the treatment of neuropathic pain. PMID- 11050136 TI - The circadian clock mutation alters sleep homeostasis in the mouse. AB - The onset and duration of sleep are thought to be primarily under the control of a homeostatic mechanism affected by previous periods of wake and sleep and a circadian timing mechanism that partitions wake and sleep into different portions of the day and night. The mouse Clock mutation induces pronounced changes in overall circadian organization. We sought to determine whether this genetic disruption of circadian timing would affect sleep homeostasis. The Clock mutation affected a number of sleep parameters during entrainment to a 12 hr light/dark (LD 12:12) cycle, when animals were free-running in constant darkness (DD), and during recovery from 6 hr of sleep deprivation in LD 12:12. In particular, in LD 12:12, heterozygous and homozygous Clock mutants slept, respectively, approximately 1 and approximately 2 hr less than wild-type mice, and they had 25 and 51% smaller increases in rapid eye movement (REM) sleep during 24 hr recovery, respectively, than wild-type mice. The effects of the mutation on sleep are not readily attributable to differential entrainment to LD 12:12 because the baseline sleep differences between genotypes were also present when animals were free-running in DD. These results indicate that genetic alterations of the circadian clock system and/or its regulatory genes are likely to have widespread effects on a variety of sleep and wake parameters, including the homeostatic regulation of sleep. PMID- 11050137 TI - Using fos imaging in the rat to reveal the anatomical extent of the disruptive effects of fornix lesions. AB - Activity of the immediate early gene c-fos was compared across hemispheres in rats with unilateral fornix lesions. To engage Fos production, rats first performed a radial arm maze task that is severely disrupted by bilateral fornix lesions. Using immunohistochemical techniques, Fos-positive cells were visualized and counted in 39 sites in both hemispheres. Fornix lesions led to a significant reduction in Fos in all ipsilateral hippocampal subfields, as well as the entorhinal cortex and most of the subicular complex. Other sites that showed reduced activity included the ipsilateral retrosplenial, anterior cingulate, and postrhinal cortices. Subcortical regions showing significant Fos decreases included the anterior thalamic nuclei, supramammillary nucleus, diagonal band of Broca, and lateral septum. Thus, the effects of fornix lesions extended beyond the hippocampal formation and included sites not directly innervated by the tract. These changes were nevertheless selective, as shown by the lack of hemispheric difference in any of the preselected control sites, the perirhinal cortex, or nucleus accumbens. Furthermore, there were no hemispheric differences in an additional group of animals with unilateral fornix lesions that were killed directly from the home cage. The location of Fos changes closely corresponded to those brain regions that when lesioned disrupt spatial working memory. Moreover, there was a correspondence between those brain regions that show increased Fos production in normal animals performing the radial arm maze task and those affected by fornix lesions. These results show that fornix transection has widespread, but selective, effects on a network of structures normally activated by spatial memory processes, with these effects extending beyond the hippocampal formation. PMID- 11050138 TI - Interleukin-1beta -induced changes in blood-brain barrier permeability, apparent diffusion coefficient, and cerebral blood volume in the rat brain: a magnetic resonance study. AB - The cytokine interleukin-1beta (IL-1beta) is implicated in a broad spectrum of CNS pathologies, in which it is thought to exacerbate neuronal loss. Here, the effects of injecting recombinant rat IL-1beta into the striatum of 3-week-old rats were followed noninvasively from 2 to 123 hr using magnetic resonance imaging and spectroscopy. Four hours after injection of IL-1beta (1 ng in 1 microliter), cerebral blood volume was significantly increased, the blood-brain barrier (BBB) became permeable to intravenously administered contrast agent between 4.5 and 5 hr, and the apparent diffusion coefficient (ADC) of brain water fell by 6 hr (5.42 +/- 0. 35 x 10(-4) mm(2)/sec treated, 7.35 +/- 0.77 x 10(-)(4) mm(2)/sec control; p < 0.001). At 24 hr the BBB was again intact, but the ADC, although partially recovered, remained depressed at both 24 and 123 hr (p < 0.03). Depleting the animals of neutrophils before IL-1beta injection prevented the BBB permeability at all time points, but the ADC was still depressed at 6 hr (6.64 +/- 0.34 x 10(-4) mm(2)/sec treated, 7.49 +/- 0.38 x 10(-4) mm(2)/sec control; p < 0.005). No changes were seen in brain metabolites using proton spectroscopy at 6 hr after IL-1beta. Intraparenchymal injection of IL-1beta caused a neutrophil-dependent transient increase in BBB permeability. The presence of neutrophils within the brain parenchyma significantly contributed to the IL-1beta-induced changes in cerebral blood volume and the ADC of brain water. However, IL-1beta apparently had a direct effect on the resident cell populations, which persisted well after all recruited leukocytes had disappeared. Thus the action of IL-1beta alone can give rise to magnetic resonance imaging visible changes that are normally attributed to alterations to cellular homeostasis. PMID- 11050139 TI - Reduction of pentylenetetrazole-induced seizure activity in awake rats by seizure triggered trigeminal nerve stimulation. AB - Stimulation of the vagus nerve has become an effective method for desynchronizing the highly coherent neural activity typically associated with epileptic seizures. This technique has been used in several animal models of seizures as well as in humans suffering from epilepsy. However, application of this technique has been limited to unilateral stimulation of the vagus nerve, typically delivered according to a fixed duty cycle, independently of whether ongoing seizure activity is present. Here, we report that stimulation of another cranial nerve, the trigeminal nerve, can also cause cortical and thalamic desynchronization, resulting in a reduction of seizure activity in awake rats. Furthermore, we demonstrate that providing this stimulation only when seizure activity begins results in more effective and safer seizure reduction per second of stimulation than with previous methods. Seizure activity induced by intraperitoneal injection of pentylenetetrazole was recorded from microwire electrodes in the thalamus and cortex of awake rats while the infraorbital branch of the trigeminal nerve was stimulated via a chronically implanted nerve cuff electrode. Continuous unilateral stimulation of the trigeminal nerve reduced electrographic seizure activity by up to 78%, and bilateral trigeminal stimulation was even more effective. Using a device that automatically detects seizure activity in real time on the basis of multichannel field potential signals, we demonstrated that seizure-triggered stimulation was more effective than the stimulation protocol involving a fixed duty cycle, in terms of the percent seizure reduction per second of stimulation. In contrast to vagus nerve stimulation studies, no substantial cardiovascular side effects were observed by unilateral or bilateral stimulation of the trigeminal nerve. These findings suggest that trigeminal nerve stimulation is safe in awake rats and should be evaluated as a therapy for human seizures. Furthermore, the results demonstrate that seizure-triggered trigeminal nerve stimulation is technically feasible and could be further developed, in conjunction with real-time seizure-predicting paradigms, to prevent seizures and reduce exposure to nerve stimulation. PMID- 11050140 TI - A cellular mechanism for the transformation of a sensory input into a motor command. AB - The initiation and control of locomotion largely depend on processing of sensory inputs. The cellular bases of locomotion have been extensively studied in lampreys where reticulospinal (RS) neurons constitute the main descending system activating and controlling the spinal locomotor networks. Ca(2+) imaging and intracellular recordings were used to study the pattern of activation of RS neurons in response to cutaneous stimulation. Pressure applied to the skin evoked a linear input/output relationship in RS neurons until a threshold level, at which a depolarizing plateau was induced, the occurrence of which was associated with the onset of swimming activity in a semi-intact preparation. The occurrence of a depolarizing plateau was abolished by blocking the NMDA receptors that are located on RS cells. Moreover, the depolarizing plateaus were accompanied by a rise in [Ca(2+)](i), and an intracellular injection of the Ca(2+) chelator BAPTA into single RS cells abolished the plateaus, suggesting that the latter are Ca(2+) dependent and rely on intrinsic properties of RS cells. The plateaus were shown to result from the activation of a Ca(2+)-activated nonselective cation current that maintains the cell in a depolarized state. It is concluded that this intrinsic property of the RS neuron is then responsible for the transformation of an incoming sensory signal into a motor command that is then forwarded to the spinal locomotor networks. PMID- 11050141 TI - Activation of ERK/MAP kinase in the amygdala is required for memory consolidation of pavlovian fear conditioning. AB - Although much has been learned about the neurobiological mechanisms underlying Pavlovian fear conditioning at the systems and cellular levels, relatively little is known about the molecular mechanisms underlying fear memory consolidation. The present experiments evaluated the role of the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) signaling cascade in the amygdala during Pavlovian fear conditioning. We first show that ERK/MAPK is transiently activated-phosphorylated in the amygdala, specifically the lateral nucleus (LA), at 60 min, but not 15, 30, or 180 min, after conditioning, and that this activation is attributable to paired presentations of tone and shock rather than to nonassociative auditory stimulation, foot shock sensitization, or unpaired tone-shock presentations. We next show that infusions of U0126, an inhibitor of ERK/MAPK activation, aimed at the LA, dose-dependently impair long term memory of Pavlovian fear conditioning but leaves short-term memory intact. Finally, we show that bath application of U0126 impairs long-term potentiation in the LA in vitro. Collectively, these results demonstrate that ERK/MAPK activation is necessary for both memory consolidation of Pavlovian fear conditioning and synaptic plasticity in the amygdala. PMID- 11050142 TI - Visual responses in monkey areas V1 and V2 to three-dimensional surface configurations. AB - The visual system uses information about the relative depth of contours and surfaces to link and segment elements of visual scenes. The integration of form and depth information was studied in areas V1 and V2 of the alert macaque. Neurons in area V2 used contextual depth information to integrate occluded contours, signal the presence of object boundaries, and segment surfaces: (1) Amodal contour completion occurs when a contour passes behind an occluder. The basis of contour completion, the facilitation of neuronal responses to stimuli located within their receptive fields (RFs) by contextual lines lying outside their RFs, was blocked by orthogonal lines intersecting the contours but was recovered when the orthogonal line was placed in the near depth plane. (2) An illusory contour will modally complete separated elements located across an isoluminant field if the elements are placed in the near depth plane. V2 neurons responded when line segments were placed outside the RF in the near depth plane and a field of uniform luminance covered the RF. (3) Texture elements within a surface will "capture" the perceived depth consistent with the disparity of the surface's boundary, even when given no disparity themselves. V2 neurons responded to the center elements of a grating as if they contained disparity, even though disparity was present only for the grating's end elements located beyond the RF borders. These results, which were more common in V2 than in V1, demonstrate a role for V2 in the three-dimensional representation of surfaces in space. PMID- 11050143 TI - Neural correlates of olfactory recognition memory in the rat orbitofrontal cortex. AB - The orbitofrontal cortex (OF) is strongly and reciprocally connected with the perirhinal (PR) and entorhinal areas of the medial temporal lobe and plays an important role in odor recognition memory. This study characterized firing patterns of single neurons in the OF of rats performing a continuous odor-guided delayed nonmatch to sample (DNMS) task. Most OF neurons fired in association with one or more task events, including the initiation of trials, the sampling of odor stimuli, and the consumption of rewards. OF neurons also exhibited sustained odor selective activity during the memory delay, and a large proportion of OF cells had odor-specific enhanced or suppressed responses on stimulus repetition. Most OF neurons were activated during several task events, or associated with complex behavioral states. The incidence of cells that fired in association with the critical match/non-match judgement was increased as the DNMS rule was learned, and was higher in OF than in perirhinal and entorhinal cortex. Furthermore, the classification of match and nonmatch trials was correlated with accuracy in performance of that judgement. These findings are consistent with the view that OF is a high order association cortex that plays a role both in the memory representations for specific stimuli and in the acquisition and application of task rules. PMID- 11050144 TI - Dopamine release and uptake dynamics within nonhuman primate striatum in vitro. AB - The putamen of the human striatum is a heterogeneous nucleus that contains the primary site of loss of dopamine (DA) in Parkinson's disease (PD). Furthermore, different functional domains of the putamen are heterogeneously susceptible to DA loss, and yet the dynamic regulation of extracellular DA concentration ([DA](o)) and comparison between domains has not been explored in the primate brain. In these studies, DA was measured in real time using fast-scan cyclic voltammetry at a carbon-fiber microelectrode in vitro in striatal sections from the common marmoset (Callithrix jacchus). [DA](o) released by a single stimulus pulse varied threefold along a ventromedial-dorsolateral axis. DA uptake was via the DA transporter (GBR12909 sensitive, desipramine insensitive). On the basis of data modeling with simulations of Michaelis-Menten kinetics, rate maximum, V(max), varied with region: both [DA](o) and V(max) were greatest in regions most vulnerable in PD. These differences were reflected in part by regional variation in DA content. [DA](o), V(max), and regional variation were two- to threefold greater than in rodent caudatoputamen. In addition, steady-state [DA](o) at physiological firing rates in primate striatum was controlled by depolarization frequency, uptake, and presynaptic autoreceptors. Furthermore, regulation of [DA](o) by these mechanisms differed significantly between limbic- and motor associated domains. These data indicate interspecies heterogeneity in striatal DA dynamics that must be considered when extrapolating behavioral and drug responses from rodent to the primate brain. Moreover, the heterogeneity demonstrated within the primate putamen in the availability and dynamic regulation of DA may be central to understanding DA function in health, cocaine abuse, and disease. PMID- 11050145 TI - Developmental changes in eye-blink conditioning and neuronal activity in the inferior olive. AB - Neuronal activity was recorded in the dorsal accessory inferior olive in infant rats during classical conditioning of the eye-blink response. The percentage and amplitude of eye-blink conditioned responses (CRs) increased as a function of age. The magnitude of the neuronal response to the unconditioned stimulus (US) decreased with age. There were also age-specific modifications of US-elicited inferior olive neuronal activity during paired trials in which a conditioned eye blink response was performed. The results indicate that the development of the conditioned eye-blink response may depend on dynamic interactions between multiple developmental processes within the eye-blink circuitry. Differences in the functional maturity of olivo-cerebellar pathways may limit the induction of plasticity in the cerebellum and thereby limit the development of eye-blink conditioned responses. PMID- 11050146 TI - Neurotrophic factor expression after CNS viral injury produces enhanced sensitivity to psychostimulants: potential mechanism for addiction vulnerability. AB - Hypothesized risk factors for psychostimulant, amphetamine, and cocaine abuse include dopamine (DA) receptor polymorphisms, HIV infection, schizophrenia, drug induced paranoias, and movement disorders; however, the molecular, cellular, and biochemical mechanisms that predispose to drug sensitivity or drive the development of addiction are incompletely understood. Using the Borna disease rat, an animal model of viral-induced encephalopathy wherein sensitivity to the locomotor and stereotypic behavioral effects of d-amphetamine and cocaine is enhanced (Solbrig et al., 1994, 1998), we identify a specific neurotrophin expression pattern triggered by striatal viral injury that increases tyrosine hydroxylase activity, an early step in DA synthesis, to produce a phenotype of enhanced amphetamine sensitivity. The reactive neurotrophin pattern provides a molecular framework for understanding how CNS viral injury, as well as other CNS adaptations producing similar growth factor activation profiles, may influence psychostimulant sensitivity. PMID- 11050147 TI - Bradykinin, but not muscarinic, inhibition of M-current in rat sympathetic ganglion neurons involves phospholipase C-beta 4. AB - Rat superior cervical ganglion (SCG) neurons express low-threshold noninactivating M-type potassium channels (I(K(M))), which can be inhibited by activation of M(1) muscarinic receptors (M(1) mAChR) and bradykinin (BK) B(2) receptors. Inhibition by the M(1) mAChR agonist oxotremorine methiodide (Oxo-M) is mediated, at least in part, by the pertussis toxin-insensitive G-protein Galpha(q) (Caulfield et al., 1994; Haley et al., 1998a), whereas BK inhibition involves Galpha(q) and/or Galpha(11) (Jones et al., 1995). Galpha(q) and Galpha(11) can stimulate phospholipase C-beta (PLC-beta), raising the possibility that PLC is involved in I(K(M)) inhibition by Oxo-M and BK. RT-PCR and antibody staining confirmed the presence of PLC-beta1, -beta2, -beta3, and -beta4 in rat SCG. We have tested the role of two PLC isoforms (PLC-beta1 and PLC-beta4) using antisense-expression constructs. Antisense constructs, consisting of the cytomegalovirus promoter driving antisense cRNA corresponding to the 3' untranslated regions of PLC-beta1 and PLC-beta4, were injected into the nucleus of dissociated SCG neurons. Injected cells showed reduced antibody staining for the relevant PLC-beta isoform when compared to uninjected cells 48 hr later. BK inhibition of I(K(M)) was significantly reduced 48 hr after injection of the PLC beta4, but not the PLC-beta1, antisense-encoding plasmid. Neither PLC-beta antisense altered M(1) mAChR inhibition by Oxo-M. These data support the conclusion of Cruzblanca et al. (1998) that BK, but not M(1) mAChR, inhibition of I(K(M)) involves PLC and extends this finding by indicating that PLC-beta4 is involved. PMID- 11050148 TI - Cloning and characterization of two splice variants of human phosphodiesterase 11A. AB - Phosphodiesterase 11A (PDE11A) is a recently identified family of cAMP and cGMP hydrolyzing enzymes. Thus far, a single splice variant designated as PDE11A1 has been reported. In this study, we identify and characterize two additional splice variants of PDE11A, PDE11A2 and PDE11A3. The full-length cDNAs are 2,141 bp for PDE11A2 and 2205 bp for PDE11A3. The ORF of PDE11A2 predicts a protein of 576 aa with a molecular mass of 65.8 kDa. The ORF of PDE11A3 predicts a protein of 684 aa with a molecular mass of 78.1 kDa. Comparison of the PDE11A2 sequence with that of PDE11A1 indicates an additional 86 aa at the N terminus of PDE11A2. Part of this sequence extends the potential cGMP binding region (GAF domain) present in PDE11A1. Compared with PDE11A2, PDE11A3 has an additional 108 N-terminal amino acids. Sequence analysis of PDE11A3 indicates the presence of another GAF domain in this region. This diversification of regulatory sequences in the N-terminal region of PDE11A splice variants suggests the interesting possibility of differential regulation of these enzymes. Recombinant PDE11A2 and -A3 proteins expressed in the Baculovirus expression system have the ability to hydrolyze both cAMP and cGMP. The K(m) values for cAMP hydrolysis are 3.3 microM and 5.7 microM for PDE11A2 and PDE11A3, respectively. The K(m) values for cGMP hydrolysis are 3.7 microM and 4.2 microM for PDE11A2 and PDE11A3, respectively. Both PDEs showed a V(max) ratio for cAMP/cGMP of approximately 1.0. PDE11A2 is sensitive to dipyridamole, with an IC(50) of 1.8 microM, and to zaprinast, with an IC(50) of 28 microM. PDE11A3 demonstrated similar pattern of inhibitor sensitivity with IC(50) values of 0.82 and 5 microM for dipyridamole and zaprinast, respectively. PMID- 11050149 TI - Nonrandom mating in Drosophila melanogaster laboratory populations derived from closely adjacent ecologically contrasting slopes at "Evolution Canyon". AB - Ecological differentiation of natural populations of Drosophila melanogaster, Drosophila simulans, and another drosophilid, Zaprionus tuberculatus, in "Evolution Canyon," Mount Carmel, Israel, is well established. The fitness complex of D. melanogaster includes oviposition temperature preferences, tolerance to high temperature, drought stress and starvation, and different longevity patterns. This remarkable differentiation has evolved despite small interslope distances (only 100-400 m), within easy dispersal distance. The differences between populations are those expected from genetic adaptation to local microclimates. How such differentiation could evolve and be maintained despite the likelihood of genetic exchange between populations is a challenging question. We hypothesized that interslope microclimatic differences caused strong differential selection for stress tolerance, accompanied by behavioral differentiation (habitat choice and reduced migration rate), reinforced by sexual isolation. Here we report highly significant mate choice by flies from different slopes of the canyon, with preference for sexual partners originating from the same slope. No preferences were found when the sexual partners belonged to different isofemale lines from the same slope. PMID- 11050150 TI - Host-parasite relatedness shown by protein fingerprinting in a brood parasitic bird. AB - Brood parasitism as an alternative female breeding tactic is particularly common in ducks, where hosts often receive eggs laid by parasitic females of the same species and raise their offspring. Herein, we test several aspects of a kin selection explanation for this phenomenon in goldeneye ducks (Bucephala clangula) by using techniques of egg albumen sampling and statistical bandsharing analysis based on resampling. We find that host and primary parasite are indeed often related, with mean r = 0.13, about as high as between first cousins. Relatedness to the host is higher in nests where a parasite lays several eggs than in those where she lays only one. Returning young females parasitize their birth nestmates (social mothers or sisters, which are usually also their genetic mothers and sisters) more often than expected by chance. Such adult relatives are also observed together in the field more often than expected and for longer periods than other females. Relatedness and kin discrimination, which can be achieved by recognition of birth nestmates, therefore play a role in these tactics and probably influence their success. PMID- 11050151 TI - Regulation of beta -catenin transformation by the p300 transcriptional coactivator. AB - The beta-catenin protein plays a critical role in embryonic development and mature tissue homeostasis through its effects on E-cadherin-mediated cell adhesion and Wnt-dependent signal transduction. In colon and other cancers, mutations of beta-catenin or the adenomatous polyposis coli (APC) tumor suppressor appear to stabilize beta-catenin and enhance its interaction with T cell factor (TCF) or lymphoid enhancer factor (Lef) transcription factors. At present, a complete picture of the means by which beta-catenin's interactions with TCF/Lef proteins contribute to neoplastic transformation is lacking. We report that the transcriptional coactivator p300 interacts with beta-catenin in vitro and in vivo and is critical for beta-catenin-mediated neoplastic transformation. p300 synergistically activates beta-catenin/TCF transcription, and their biochemical association requires the CH1 domain of p300 and a region of beta-catenin that includes its NH(2)-terminal transactivation domain and the first two armadillo repeats. Lowering of cellular p300 levels by using a ribozyme directed against p300 reduced TCF transcriptional activity and inhibited the neoplastic growth properties of a beta-catenin-transformed rat epithelial cell line and a human colon carcinoma line with a beta-catenin mutation. These findings demonstrate a critical role for p300 in beta-catenin/TCF transcription and in cancers arising from defects in beta-catenin regulation. PMID- 11050152 TI - 2'-Hydroxylation of nicotine by cytochrome P450 2A6 and human liver microsomes: formation of a lung carcinogen precursor. AB - Smokers or people undergoing nicotine replacement therapy excrete approximately 10% of the nicotine dose as 4-oxo-4-(3-pyridyl)butanoic acid (keto acid) and 4 hydroxy-4-(3-pyridyl)butanoic acid (hydroxy acid). Previously, these acids were thought to arise by secondary metabolism of the major nicotine metabolite cotinine, but our data did not support this mechanism. Therefore, we hypothesized that nicotine is metabolized by 2'-hydroxylation, which would ultimately yield keto acid and hydroxy acid as urinary metabolites. This pathway had not been established previously in mammalian systems and is potentially significant because the product of nicotine 2'-hydroxylation, 4-(methylamino)-1-(3-pyridyl)-1 butanone (aminoketone), can be converted to the potent tobacco-specific lung carcinogen, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. Incubation of nicotine with cytochrome P450 2A6 and cofactors did indeed produce aminoketone, which was identified as its N-benzoyl derivative by GC-MS. The rate was 11% of that of cotinine production. Incubation of human liver microsomes with nicotine gave keto acid by using aminoketone as an intermediate; keto acid was not formed from cotinine. In 10 human liver samples, rates of formation of keto acid were 5.7% of those of cotinine and production of these metabolites correlated. These results provide definitive evidence for mammalian 2'-hydroxylation of nicotine and elucidate a pathway by which endogenous formation of 4-(methylnitrosamino)-1 (3-pyridyl)-1-butanone could occur in humans. PMID- 11050154 TI - Impact of a Permo-Carboniferous high O2 event on the terrestrial carbon cycle. AB - Independent models predicting the Phanerozoic (past 600 million years) history of atmospheric O(2) partial pressure (pO(2)) indicate a marked rise to approximately 35% in the Permo-Carboniferous, around 300 million years before present, with the strong potential for altering the biogeochemical cycling of carbon by terrestrial ecosystems. This potential, however, would have been modified by the prevailing atmospheric pCO(2) value. Herein, we use a process-based terrestrial carbon cycle model forced with a late Carboniferous paleoclimate simulation to evaluate the effects of a rise from 21 to 35% pO(2) on terrestrial biosphere productivity and assess how this response is modified by current uncertainties in the prevailing pCO(2) value. Our results indicate that a rise in pO(2) from 21 to 35% during the Carboniferous reduced global terrestrial primary productivity by 20% and led to a 216-Gt (1 Gt = 10(12) kg) C reduction in the vegetation and soil carbon storage, in an atmosphere with pCO(2) = 0.03%. However, in an atmosphere with pCO(2) = 0.06%, the CO(2) fertilization effect is larger than the cost of photorespiration, and ecosystem productivity increases leading to the net sequestration of 117 Gt C into the vegetation and soil carbon reservoirs. In both cases, the effects result from the strong interaction between pO(2), pCO(2), and climate in the tropics. From this analysis, we deduce that a Permo-Carboniferous rise in pO(2) was unlikely to have exerted catastrophic effects on ecosystem productivity (with pCO(2) = 0.03%), and if pCO(2) levels at this time were >0.04%, the water-use efficiency of land plants may even have improved. PMID- 11050153 TI - Long-term regulation of neuronal high-affinity glutamate and glutamine uptake in Aplysia. AB - An increase in transmitter release accompanying long-term sensitization and facilitation occurs at the glutamatergic sensorimotor synapse of Aplysia. We report that a long-term increase in neuronal Glu uptake also accompanies long term sensitization. Synaptosomes from pleural-pedal ganglia exhibited sodium dependent, high-affinity Glu transport. Different treatments that induce long term enhancement of the siphon-withdrawal reflex, or long-term synaptic facilitation increased Glu uptake. Moreover, 5-hydroxytryptamine, a treatment that induces long-term facilitation, also produced a long-term increase in Glu uptake in cultures of sensory neurons. The mechanism for the increase in uptake is an increase in the V(max) of transport. The long-term increase in Glu uptake appeared to be dependent on mRNA and protein synthesis, and transport through the Golgi, because 5,6-dichlorobenzimidazole riboside, emetine, and brefeldin A inhibited the increase in Glu uptake. Also, injection of emetine and 5,6 dichlorobenzimidazole into Aplysia prevented long-term sensitization. Synthesis of Glu itself may be regulated during long-term sensitization because the same treatments that produced an increase in Glu uptake also produced a parallel increase in Gln uptake. These results suggest that coordinated regulation of a number of different processes may be required to establish or maintain long-term synaptic facilitation. PMID- 11050155 TI - Functional mosaic organization of mouse olfactory receptor neurons. AB - In contrast to rapid progress in the molecular biology of olfaction, there are few physiological data to characterize the odor response properties of different populations of olfactory receptor neurons (ORNs) and their spatial distributions across the epithelium, which is essential for understanding the coding mechanisms underlying odor discrimination and recognition. We have tested the hypothesis that the ORNs are arranged in a functional mosaic, using an intact epithelial preparation from the mouse, in which odor responses of many ORNs in situ can be monitored simultaneously with calcium imaging techniques. ORNs responding to a given odor were widely distributed across epithelium and intermingled with ORNs responding to other odors. Tight clusters of ORNs responding to the same odor were observed. For a given odor, more ORNs were recruited when the concentration was increased. ORNs were able to distinguish between pairs of enantiomers by showing distinct but somewhat overlapping patterns. The results provide evidence regarding the response spectra of ORNs in situ, supporting the combinatorial coding of odor quality and intensity by different ORN subsets. PMID- 11050156 TI - The classical progesterone receptor mediates Xenopus oocyte maturation through a nongenomic mechanism. AB - Xenopus laevis oocytes are physiologically arrested at G(2) of meiosis I. Resumption of meiosis, or oocyte maturation, is triggered by progesterone. Progesterone-induced Xenopus oocyte maturation is mediated via an extranuclear receptor and is independent of gene transcription. The identity of this extranuclear oocyte progesterone receptor (PR), however, has remained a longstanding problem. We have isolated the amphibian homologue of human PR from a Xenopus oocyte cDNA library. The cloned Xenopus progesterone receptor (xPR) functioned in heterologous cells as a progesterone-regulated transcription activator. However, endogenous xPR was excluded from the oocyte nucleus and instead appeared to be a cytosolic protein not associated with any membrane structures. Injection of xPR mRNA into Xenopus oocytes accelerated the progesterone-induced oocyte maturation and reduced the required concentrations of progesterone. In enucleated oocytes, xPR accelerated the progesterone-induced mitogen-activated protein kinase activation. These data suggest that xPR is the long sought after Xenopus oocyte receptor responsible for progesterone-induced oocyte maturation. PMID- 11050157 TI - Combinatorial peptide libraries reveal the ligand-binding mechanism of the oligopeptide receptor OppA of Lactococcus lactis. AB - The oligopeptide transport system (Opp) of Lactococcus lactis has the unique capacity to mediate the transport of peptides from 4 up to at least 18 residues. The substrate specificity of this binding protein-dependent ATP-binding cassette transporter is determined mainly by the receptor protein OppA. To study the specificity and ligand-binding mechanism of OppA, the following strategy was used: (i) OppA was purified and anchored via the lipid moiety to the surface of liposomes; (ii) the proteoliposomes were used in a rapid filtration-based binding assay with radiolabeled nonameric bradykinin as a reporter peptide; and (iii) combinatorial peptide libraries were used to determine the specificity and selectivity of OppA. The studies show that (i) OppA is able to bind peptides up to at least 35 residues, but there is a clear optimum in affinity for nonameric peptides; (ii) the specificity for nonameric peptides is not equally distributed over the whole peptide, because positions 4, 5, and 6 in the binding site are more selective; and (iii) the differences in affinity for given side chains is relatively small, but overall hydrophobic residues are favored-whereas glycine, proline, and negatively charged residues lower the binding affinity. The data indicate that not only the first six residues (enclosed by the protein) but also the C-terminal three residues interact in a nonopportunistic manner with (the surface of) OppA. This binding mechanism is different from the one generally accepted for receptors of ATP-binding cassette-transporter systems. PMID- 11050158 TI - A dibasic motif involved in parathyroid hormone-induced down-regulation of the type IIa NaPi cotransporter. AB - Type II NaPi cotransporters are expressed in the apical membrane of P(i) (re)absorbing epithelia: the type IIa in renal proximal tubule and the type IIb in small intestine. Parathyroid hormone (PTH) leads to a retrieval from the apical membrane of the type IIa NaPi cotransporter. The type IIa cotransporter is also expressed in opossum kidney (OK) cells, and its expression is under the control of PTH. In the present study, we identified the molecular "domains" involved in the PTH-induced retrieval of the type IIa NaPi cotransporter. Wild type mouse type IIa (mIIa) and type IIb (mIIb) as well as several mIIa-mIIb chimeras and site-directed mutants were fused to the enhanced green fluorescent protein and transfected into OK cells. We found that mIIa but not mIIb was internalized and degraded after incubation with 1-34 (or 3-34) PTH. Using chimeras, we found that the N and C termini were not required in this effect, whereas a "domain" located between residues 216 and 658 seemed to be necessary. This region contains two putative intracellular loops with highly conserved sequences between mIIa and mIIb; in the last intracellular loop, two charged amino acids of type IIa (K(503)R(504)) are replaced by uncharged residues in type IIb (N(520)I(521)). We generated two mutants in which these residues were interchanged: mIIaNI and mIIbKR. Similarly to mIIa, the mIIbKR mutant was endocytosed in response to 1-34 PTH; in contrast, mIIaNI behaved as mIIb and was not internalized. In conclusion, a dibasic amino acid motif (K(503)R(504)) located in the last intracellular loop of the type IIa NaPi cotransporter is essential for its PTH-induced retrieval. PMID- 11050159 TI - Bestrophin, the product of the Best vitelliform macular dystrophy gene (VMD2), localizes to the basolateral plasma membrane of the retinal pigment epithelium. AB - Best vitelliform macular dystrophy is a dominantly inherited, early onset, macular degenerative disease that exhibits some histopathologic similarities to age-related macular degeneration. Although the vitelliform lesion is common in the fundus of individuals with Best disease, diagnosis is based on a reduced ratio of the light peak to dark trough in the electrooculogram. Recently, the VMD2 gene on chromosome 11q13, encoding the protein bestrophin, was identified. The function of bestrophin is unknown. To facilitate studies of bestrophin, we produced both rabbit polyclonal and mouse monoclonal antibodies that proved useful for Western blotting, immunoprecipitation, and immunocytochemistry. To characterize bestrophin, we initially probed the retinal pigment epithelium (RPE) derived cell lines ARPE-19, D407, and RPE-J. All of the cell lines expressed bestrophin mRNA by reverse transcription-PCR, but not on Western blots. Bestrophin in human RPE partitioned in the detergent phase during Triton X-114 extraction and could be modified by biotin in intact cells, indicative of a plasma membrane localization. Immunocytochemical staining of macaque and porcine eyes indicated that bestrophin is localized at the basolateral plasma membrane of RPE cells. When expressed in RPE-J cells by adenovirus-mediated gene transfer, bestrophin again was determined by confocal microscopy and cell surface biotinylation to be a basolateral plasma membrane protein. The basolateral plasma membrane localization of bestrophin suggests the possibility that bestrophin plays a role in generating the altered electrooculogram of individuals with Best disease. PMID- 11050160 TI - Active acetyl-CoA synthase from Clostridium thermoaceticum obtained by cloning and heterologous expression of acsAB in Escherichia coli. AB - Acetyl-CoA synthase from Clostridium thermoaceticum (ACS(Ct)) is an alpha(2)beta(2) tetramer containing two novel Ni-X-Fe(4)S(4) active sites (the A and C clusters) and a standard Fe(4)S(4) cluster (the B cluster). The acsA and acsB genes encoding the enzyme were cloned into Escherichia coli strain JM109 and overexpressed at 37(o)C under anaerobic conditions with Ni supplementation. The isolated recombinant His-tagged protein (AcsAB) exhibited characteristics essentially indistinguishable from those of ACS(Ct), from which Ni had been removed from the A cluster. AcsAB migrated through nondenaturing electrophoretic gels as a single band and contained a 1:1 molar ratio of subunits and 1.0-1.6 Ni/alphabeta and 14-22 Fe/alphabeta. AcsAB exhibited 100-250 units/mg CO oxidation activity but no CO/acetyl-CoA exchange activity. Electronic absorption spectra of thionin-oxidized and CO-reduced AcsAB were similar to those of ACS(Ct), with features typical of redox-active Fe(4)S(4) clusters. Partially oxidized and CO-reduced AcsAB exhibited EPR signals with g values and low spin intensities indistinguishable from those of the B(red) state of the B cluster and the C(red1) and C(red2) states of the C cluster of ACS(Ct). Upon overnight exposure to NiCl(2), the resulting recombinant enzyme (ACS(Ec)) developed 0. 06 0.25 units/mg exchange activity. The highest of these values is typical of fully active ACS(Ct). When reduced with CO, ACS(Ec) exhibited an EPR signal indistinguishable from the NiFeC signal of Ni-replete ACS(Ct). Variability of activities and signal intensities were observed among different preparations. Issues involving the assembly of these metal centers in E. coli are discussed. PMID- 11050161 TI - Amplification of dopaminergic signaling by a positive feedback loop. AB - Dopamine and cAMP-regulated phosphoprotein of M(r) 32,000 (DARPP-32) plays an obligatory role in most of the actions of dopamine. In resting neostriatal slices, cyclin-dependent kinase 5 (Cdk5) phosphorylates DARPP-32 at Thr-75, thereby reducing the efficacy of dopaminergic signaling. We report here that dopamine, in slices, and acute cocaine, in whole animals, decreases the state of phosphorylation of striatal DARPP-32 at Thr-75 and thereby removes this inhibitory constraint. This effect of dopamine is achieved through dopamine D1 receptor-mediated activation of cAMP-dependent protein kinase (PKA). The activated PKA, by decreasing the state of phosphorylation of DARPP-32-Thr-75, de inhibits itself. Dopamine D2 receptor stimulation has the opposite effect. The ability of activated PKA to reduce the state of phosphorylation of DARPP-32-Thr 75 is apparently attributable to increased protein phosphatase-2A activity, with Cdk5 being unaffected. Together, these results indicate that via positive feedback mechanisms, Cdk5 signaling and PKA signaling are mutually antagonistic. PMID- 11050162 TI - Increased p53 mutation load in nontumorous human liver of wilson disease and hemochromatosis: oxyradical overload diseases. AB - Hemochromatosis and Wilson disease (WD), characterized by the excess hepatic deposition of iron and copper, respectively, produce oxidative stress and increase the risk of liver cancer. Because the frequency of p53 mutated alleles in nontumorous human tissue may be a biomarker of oxyradical damage and identify individuals at increased cancer risk, we have determined the frequency of p53 mutated alleles in nontumorous liver tissue from WD and hemochromatosis patients. When compared with the liver samples from normal controls, higher frequencies of G:C to T:A transversions at codon 249 (P < 0.001) and C:G to A:T transversions and C:G to T:A transitions at codon 250 (P < 0.001 and P < 0.005) were found in liver tissue from WD cases, and a higher frequency of G:C to T:A transversions at codon 249 (P < 0.05) also was found in liver tissue from hemochromatosis cases. Sixty percent of the WD and 28% of hemochromatosis cases also showed a higher expression of inducible nitric oxide synthase in the liver, which suggests nitric oxide as a source of increased oxidative stress. A high level of etheno-DNA adducts, formed from oxyradical-induced lipid peroxidation, in liver from WD and hemochromatosis patients has been reported previously. Therefore, we exposed a wild-type p53 TK-6 lymphoblastoid cell line to 4-hydroxynonenal, an unsaturated aldehyde involved in lipid peroxidation, and observed an increase in G to T transversions at p53 codon 249 (AGG to AGT). These results are consistent with the hypothesis that the generation of oxygen/nitrogen species and unsaturated aldehydes from iron and copper overload in hemochromatosis and WD causes mutations in the p53 tumor suppressor gene. PMID- 11050163 TI - Formation of high molecular weight complexes of mutant Cu, Zn-superoxide dismutase in a mouse model for familial amyotrophic lateral sclerosis. AB - Deposition of aggregated protein into neurofilament-rich cytoplasmic inclusion bodies is a common cytopathological feature of neurodegenerative disease. How-or indeed whether-protein aggregation and inclusion body formation cause neurotoxicity are presently unknown. Here, we show that the capacity of superoxide dismutase (SOD) to aggregate into biochemically distinct, high molecular weight, insoluble protein complexes (IPCs) is a gain of function associated with mutations linked to autosomal dominant familial amyotrophic lateral sclerosis. SOD IPCs are detectable in spinal cord extracts from transgenic mice expressing mutant SOD several months before inclusion bodies and motor neuron pathology are apparent. Sequestration of mutant SOD into cytoplasmic inclusion bodies resembling aggresomes requires retrograde transport on microtubules. These data indicate that aggregation and inclusion body formation are mechanistically and temporally distinct processes. PMID- 11050164 TI - Condensed complexes, rafts, and the chemical activity of cholesterol in membranes. AB - Epifluorescence microscopy studies of mixtures of phospholipids and cholesterol at the air-water interface often exhibit coexisting liquid phases. The properties of these liquids point to the formation of "condensed complexes" between cholesterol and certain phospholipids, such as sphingomyelin. It is found that monolayers that form complexes can incorporate a low concentration of a ganglioside G(M1). This glycolipid is visualized by using a fluorescently labeled B subunit of cholera toxin. Three coexisting liquid phases are found by using this probe together with a fluorescent phospholipid probe. The three liquid phases are identified as a phospholipid-rich phase, a cholesterol-rich phase, and a condensed complex-rich phase. The cholera toxin B labeled ganglioside G(M1) is found exclusively in the condensed complex-rich phase. Condensed complexes are likely present in animal cell membranes, where they should facilitate the formation of specialized domains such as rafts. Condensed complexes also have a major effect in determining the chemical activity of cholesterol. It is suggested that this chemical activity plays an essential role in the regulation of cholesterol biosynthesis. Gradients in the chemical activity of cholesterol should likewise govern the rates and direction of intracellular intermembrane cholesterol transport. PMID- 11050165 TI - Peripheral Golgi protein GRASP65 is a target of mitotic polo-like kinase (Plk) and Cdc2. AB - Cell division is characterized by orchestrated events of chromosome segregation, distribution of cellular organelles, and the eventual partitioning and separation of the two daughter cells. Mitotic kinases, including polo-like kinases (Plk), influence multiple events in mitosis. In yeast two-hybrid screens using mammalian Plk C-terminal domain baits, we have identified Golgi peripheral protein GRASP65 (Golgi reassembly stacking protein of 65 kDa) as a Plk-binding protein. GRASP65 appears to function in the postmitotic reassembly of Golgi stacks. In this report we demonstrate binding between Plk and GRASP65 and provide in vitro and in vivo evidence that Plk is a GRASP65 kinase. Moreover, we show that Cdc2 can also phosphorylate GRASP65. In addition, we present data which support the observation that the conserved C terminus of Plk is important for its function. Deletion or frameshift mutations in the conserved C-terminal domain of Plk greatly diminish its ability to phosphorylate GRASP65. These and previous findings suggest that phosphorylation of Golgi components by mitotic kinases may regulate mechanisms of Golgi inheritance during cell division. PMID- 11050166 TI - Cytotoxic T lymphocyte antigen-4 (CTLA-4) regulates the size, reactivity, and function of a primed pool of CD4+ T cells. AB - We examined how cytotoxic T lymphocyte antigen-4 (CTLA-4) regulates heterogeneous CD4(+) T cell responses by using experimental autoimmune encephalomyelitis (EAE), a CD4(+) T cell-mediated disease that is subject to regulation by CTLA-4. Disease incidence and severity were used as measures of in vivo CD4(+) T cell responses. The frequency, cytokine production, and reactivity of primed T cells were determined from animals immunized with proteolipid protein (PLP)-139-151 (disease agonist), PLP-Q (disease antagonist), or both peptides, and treated with control or anti-CTLA-4 antibody to analyze the responding population. CTLA-4 blockade exacerbated disease in PLP-139-151-primed animals and overcame disease antagonism in coimmunized animals, but did not permit disease induction in PLP-Q-primed animals. Experimental autoimmune encephalomyelitis enhancement was associated with increased frequencies of cytokine-producing cells and increased ratios of IFN-gamma to IL-4 secretors responsive to PLP-139-151. Priming with PLP-Q elicited IL-4 and IL-2, but not IFN-gamma secretors cross-reactive with PLP-139 151. Strikingly, CTLA-4 blockade was found to decrease rather than increase the frequencies of cross-reactive IL-4 and IL-2 secretors. Thus, CTLA-4 engagement limits the size, but increases the breadth, of reactivity of a primed pool of CD4(+) T cells, consequently regulating its function. PMID- 11050167 TI - SOS mutator activity: unequal mutagenesis on leading and lagging strands. AB - A major pathway of mutagenesis in Escherichia coli is mediated by the inducible SOS response. Current models of SOS mutagenesis invoke the interaction of RecA and UmuD'(2)C proteins with a stalled DNA replication complex at sites of DNA lesions or poorly extendable terminal mismatches, resulting in an (error-prone) continuation of DNA synthesis. The precise mechanisms of SOS-mediated lesion bypass or mismatch extension are not known. Here, we have studied mutagenesis on the E. coli chromosome in recA730 strains. In recA730 strains, the SOS system is expressed constitutively, resulting in a spontaneous mutator effect (SOS mutator) because of reduced replication fidelity. We investigated whether during SOS mutator activity replication fidelity might be altered differentially in the leading and lagging strand of replication. Pairs of recA730 strains were constructed differing in the orientation of the lac operon relative to the origin of replication. The strains were also mismatch-repair defective (mutL) to facilitate scoring of replication errors. Within each pair, a given lac sequence is replicated by the leading-strand machinery in one orientation and by the lagging-strand machinery in the other orientation. Measurements of defined lac mutant frequencies in such pairs revealed large differences between the two orientations. Furthermore, in all cases, the frequency bias was the opposite of that seen in normal cells. We suggest that, for the lacZ target used in this study, SOS mutator activity operates with very different efficiency in the two strands. Specifically, the lagging strand of replication appears most susceptible to the SOS mutator effect. PMID- 11050168 TI - Vigilin binding selectively inhibits cleavage of the vitellogenin mRNA 3' untranslated region by the mRNA endonuclease polysomal ribonuclease 1. AB - In Xenopus, estrogen induces the stabilization of vitellogenin mRNA and the destabilization of albumin mRNA. These processes correlate with increased polysomal activity of a sequence-selective mRNA endonuclease, PMR-1, and a hnRNP K homology-domain RNA-binding protein, vigilin. Vigilin binds to a region of the vitellogenin mRNA 3'-untranslated region (3'-UTR) implicated in estrogen-mediated stabilization. The vigilin-binding site in the vitellogenin B1 mRNA 3'-UTR contains two consensus PMR-1 cleavage sites. The availability of purified PMR-1 and recombinant vigilin made it possible to test the hypothesis that RNA-binding proteins interact with cis-acting elements to stabilize target mRNAs by blocking cleavage by site-specific mRNA endonucleases. Vigilin binds to the vitellogenin mRNA 3'-UTR site with at least 30-fold higher affinity than it exhibits for the albumin mRNA segment containing the mapped PMR-1 cleavage sites. This differential binding affinity correlates with differential in vitro susceptibility of the protein-RNA complexes to cleavage by PMR-1. Whereas recombinant vigilin has no detectable protective effect on PMR-1 cleavage of albumin mRNA, it retards in vitro cleavage of the vitellogenin mRNA 3'-UTR by purified PMR-1. The PMR-1 sites in the vitellogenin mRNA 3'-UTR are functional because they are readily cleaved in vitro by purified PMR-1. These results provide direct evidence for differential susceptibility to endonuclease-mediated mRNA decay resulting from the differential affinity of a RNA-binding protein for cis-acting stability determinants. PMID- 11050169 TI - Role of C/EBP homologous protein (CHOP-10) in the programmed activation of CCAAT/enhancer-binding protein-beta during adipogenesis. AB - Hormone induction of growth-arrested preadipocytes triggers mitotic clonal expansion followed by expression of CCAAT/enhancer-binding protein (C/EBP)alpha and differentiation into adipocytes. The order of these events is critical because C/EBPalpha is antimitotic and its expression prematurely would block the mitotic clonal expansion required for differentiation. C/EBPbeta, a transcriptional activator of the C/EBPalpha gene, is expressed early in the differentiation program, but lacks DNA-binding activity and fails to localize to centromeres until preadipocytes traverse the G(1)-S checkpoint of mitotic clonal expansion. Evidence is presented that dominant-negative CHOP-10 expressed by growth-arrested preadipocytes transiently sequesters C/EBPbeta by heterodimerization. As preadipocytes reach S phase, CHOP-10 is down-regulated, apparently releasing C/EBPbeta from inhibitory constraint and allowing transactivation of the C/EBPalpha gene. In support of these findings, up regulation of CHOP-10 with the protease inhibitor N-acetyl-Leu-Leu-norleucinal prevents activation of C/EBPbeta, expression of C/EBPalpha, and adipogenesis. PMID- 11050170 TI - Sequential repression and activation of the CCAAT enhancer-binding protein-alpha (C/EBPalpha ) gene during adipogenesis. AB - CCAAT enhancer-binding protein-alpha (C/EBPalpha) functions as a pleiotropic transcriptional activator of adipocyte genes during adipogenesis. Nuclear factor C/EBP undifferentiated protein (CUP), an isoform of activator protein-2alpha (AP 2alpha), binds to repressive elements in the C/EBPalpha gene promoter, silencing the gene until late in the differentiation program. The CUP regulatory element overlaps a Sp (GT-box) element in the promoter to which Sp3 (or Sp1) can bind. Binding by Sp3 or Sp1 and CUP/AP2-alpha is mutually exclusive. Sp3 is a strong transcriptional activator of the C/EBPalpha gene promoter in 3T3-L1 preadipocytes and Schneider cells, this activation being repressed by CUP/AP-2alpha. Sp3 is expressed throughout differentiation, whereas CUP/AP-2alpha, which is expressed only by preadipocytes, is down-regulated during differentiation coincident with transcription of the C/EBPalpha gene. Thus, CUP/AP-2alpha delays access of Sp3 to the Sp regulatory element, preventing premature expression of C/EBPalpha and thereby interference by C/EBPalpha (which is antimitotic) with mitotic clonal expansion, an essential early event in the differentiation program. PMID- 11050171 TI - The Arabidopsis aldehyde oxidase 3 (AAO3) gene product catalyzes the final step in abscisic acid biosynthesis in leaves. AB - Abscisic acid (ABA) is a plant hormone involved in seed development and germination and in responses to various environmental stresses. The last step of ABA biosynthesis involves oxidation of abscisic aldehyde, and aldehyde oxidase (EC ) is thought to catalyze this reaction. An aldehyde oxidase isoform, AOdelta, encoded by AAO3, one of four Arabidopsis aldehyde oxidase genes (AAO1, AAO2, AAO3, and AAO4), is the most likely candidate for the enzyme, because it can efficiently catalyze the oxidation of abscisic aldehyde to ABA. Here, we report the isolation and characterization of an ABA-deficient Arabidopsis mutant that maps at the AAO3 locus. The mutant exhibits a wilty phenotype in rosette leaves, but seed dormancy is not affected. ABA levels were significantly reduced in the mutant leaves, explaining the wilty phenotype in rosettes, whereas the level in the mutant seeds was less reduced. No AOdelta activity could be detected in the rosette leaves of the mutant. Sequence data showed that the mutant contains a G to A substitution in the AAO3 gene. The mutation causes incorrect splicing of the ninth intron of AAO3 mRNA. We thus conclude that the ABA-deficient mutant is impaired in the AAO3 gene and that the gene product, AOdelta, is an aldehyde oxidase that catalyzes the last step of ABA biosynthesis in Arabidopsis, specifically in rosette leaves. Other aldehyde oxidases may be involved in ABA biosynthesis in other organs. PMID- 11050172 TI - Synthesis and characterization of selenotrisulfide-derivatives of lipoic acid and lipoamide. AB - Thiol-containing compounds, such as glutathione and cysteine, react with selenite under specific conditions to form selenotrisulfides. Previous studies have focused on isolation and characterization of intermolecular selenotrisulfides. This study describes the preparation and characterization of intramolecular selenotrisulfide derivatives of lipoic acid and lipoamide. These derivatives, after separation from other reaction products by reverse-phase HPLC, exhibit an absorbance maximum at 288 nm with an extinction coefficient of 1,500 M(-1) small middle dotcm(-1). The selenotrisulfide derivative of lipoic acid was significantly stable at or below pH 8.0 in contrast to several other previously studied selenotrisulfides. Mass spectral analysis of the lipoic acid and lipoamide derivatives confirmed both the expected molecular weights and also the presence of a single atom of selenium as revealed by its isotopic distribution. The selenotrisulfide derivative of lipoic acid was found to serve as an effective substrate for recombinant human thioredoxin reductase as well as native rat thioredoxin reductase in the presence of NADPH. Likewise, the lipoamide derivative was efficiently reduced by NADH-dependent bovine lipoamide dehydrogenase. The significant in vitro stability of these intramolecular selenotrisulfide derivatives of lipoic acid can serve as an important asset in the study of such selenium adducts as model selenium donor compounds for selenophosphate biosynthesis and as rate enhancement effectors in various redox reactions. PMID- 11050173 TI - Linking immune-mediated arterial inflammation and cholesterol-induced atherosclerosis in a transgenic mouse model. AB - Arterial inflammatory responses are thought to be a significant component of atherosclerotic disease. We describe here, using a transgenic approach, the mutual perpetuation of immune-mediated arterial inflammation and cholesterol induced atherosclerosis. Mice expressing the bacterial transgene beta galactosidase exclusively in cardiomyocytes and in smooth muscle cells in lung arteries and the aorta (SM-LacZ), and hypercholesterolemic apolipoprotein E deficient SM-LacZ mice (SM-LacZ/apoE(-/-)) developed myocarditis and arteritis after immunization with dendritic cells presenting a beta-galactosidase-derived immunogenic peptide. Hypercholesterolemia amplified acute arteritis and perpetuated chronic arterial inflammation in SM-LacZ/apoE(-/-) mice, but had no major impact on acute myocarditis or the subsequent development of dilated cardiomyopathy. Conversely, arteritis significantly accelerated cholesterol induced atherosclerosis. Taken together, these data demonstrate that the linkage of immune-mediated arteritis and hypercholesterolemia favors initiation and maintenance of atherosclerotic lesion formation. Therapeutic strategies to prevent or disrupt such self-perpetuating vicious circles may be crucial for the successful treatment of atherosclerosis. PMID- 11050174 TI - Endogenously expressed estrogen receptor and coactivator AIB1 interact in MCF-7 human breast cancer cells. AB - Coactivators are believed to mediate estrogen-induced gene responses via interaction with estrogen receptors (ER). Currently, a major challenge is to determine the importance of each coactivator in a specific cell type and promoter context in response to a particular ligand. The potential of ER to interact with a growing list of coactivators has been shown in a variety of in vitro and gene transfer assays, yet very few data have demonstrated the interaction of endogenous coactivators with ER in intact cells. We report here a ligand-specific interaction of endogenous human ER (hER) and the AIB1 coactivator in MCF-7 human breast cancer cells by using immunoprecipitation analyses. Complexes between endogenously expressed hER and AIB1 were detected in estradiol-treated cells and to a much lesser extent in cells treated with the partial agonist, monohydroxytamoxifen. We were unable to detect an hER-SRC-1 complex in our immunoprecipitations from MCF-7 cells. The in vitro-binding affinity for mouse ER interaction with AIB1 was estimated to be 40-120 nM. We conclude that AIB1 is a major coactivator for hER in MCF-7 human breast cancer cells. PMID- 11050175 TI - Dimerization and N-terminal domain proximity underlie the function of the molecular chaperone heat shock protein 90. AB - Heat shock protein (hsp)90 functions in a complex chaperoning pathway where its activity is modulated by ATP and by interaction with several co-chaperones. One co-chaperone, p23, binds selectively to the ATP-bound state of hsp90. However, the isolated ATP-binding domain of hsp90 does not bind p23. In an effort to identify the p23-binding domain, we have constructed a series of hsp90 deletion mutants fused with glutathione-S-transferase (GST). Full-length GST-hsp90 is able to bind p23, and also, to chaperone assembly of progesterone receptor complexes. Truncations from the C terminus of GST-hsp90 reveal a C-terminal boundary for the p23-binding domain at approximately residue 490. This fragment contains, in order, the ATP-binding domain, a highly charged region, and 203 residues beyond the charged region. p23 binding is unaffected by deletion of the charged region, indicating that two noncontiguous regions of hsp90 are involved in p23 binding. These regions are only effective when hsp90 is in a dimeric state as shown by loss of p23 binding upon removal of GST or as shown by use of FK506-binding protein12-hsp90 constructs that form dimers and bind p23 only in the presence of a bivalent drug. Thus, p23 binding requires an hsp90 dimer with close proximity between N-terminal regions of hsp90 and a conformation specified by ATP. PMID- 11050176 TI - Timing of hepatocyte entry into DNA synthesis after partial hepatectomy is cell autonomous. AB - After surgical removal of two-thirds of the liver, remaining hepatocytes replicate and restore hepatic mass within 2 weeks. This process must be initiated by signals extrinsic to the hepatocyte, but it remains unclear whether subsequent events leading to DNA synthesis (S phase) are regulated by circulating or locally produced growth factors (a noncell autonomous response), or by a program intrinsic to the hepatocyte itself (a cell autonomous response). To identify the type of mechanism regulating passage to S, we exploited the difference between rat and mouse hepatocytes in the timing of DNA synthesis after partial hepatectomy, which peaks 12-16 h earlier posthepatectomy in rat compared with mouse. Four groups of animals received two-thirds partial hepatectomies: rats, mice, mice with chimeric livers composed of both transplanted rat hepatocytes and endogenous mouse hepatocytes, and mice with chimeric livers composed of both transplanted and endogenous mouse hepatocytes. Following two-thirds partial hepatectomy, both donor and endogenous hepatocytes in mouse/mouse chimeric livers displayed kinetics of DNA synthesis characteristic of the mouse, indicating that transplantation per se did not affect the response to subsequent partial hepatectomy. In contrast, rat hepatocytes in chimeric mouse livers displayed rat kinetics despite their presence in a mouse host. Thus, factors intrinsic to the hepatocyte must regulate the timing of entry into DNA synthesis. This result defines the process as cell autonomous and suggests that locally or distantly produced cytokines or growth factors may have a permissive but not an instructive role in progression to S. PMID- 11050177 TI - Beryllium presentation to CD4+ T cells underlies disease-susceptibility HLA-DP alleles in chronic beryllium disease. AB - Chronic beryllium disease results from beryllium exposure in the workplace and is characterized by CD4(+) T cell-mediated inflammation in the lung. Susceptibility to this disease is associated with particular HLA-DP alleles. We isolated beryllium-specific T cell lines from the lungs of affected patients. These CD4(+) T cell lines specifically responded to beryllium in culture in the presence of antigen-presenting cells that expressed class II MHC molecules HLA-DR, -DQ, and DP. The response to beryllium was nearly completely and selectively blocked by mAb to HLA-DP. Additional studies showed that only certain HLA-DP alleles allowed presentation of beryllium. Overall, the DP alleles that presented beryllium to disease-specific T cell lines match those implicated in disease susceptibility, providing a mechanism for this association. Based on amino acid residues shared by these restricting and susceptibility DP alleles, our results provide insight into the residues of the DP beta-chain required for beryllium presentation. PMID- 11050178 TI - Genetic mosaic analysis based on Cre recombinase and navigated laser capture microdissection. AB - Defining molecular interactions that occur at the interface between "normal" and "abnormal" cell populations represents an important but often underexplored aspect of the pathogenesis of diseases with focal origins. Here, we illustrate an approach for conducting such analyses based on mosaic patterns of Cre recombinase expression in the adult mouse intestinal epithelium. Transgenic mice were generated that express Cre in the stem cell niche of crypts located in specified regions of their intestine. Some of these mice were engineered to allow for doxycycline-inducible Cre expression. Recombination in all pedigrees was mosaic: Cre-expressing crypts that supported recombination in all of their active multipotent stem cells were located adjacent to "control" crypts that did not express Cre at detectable levels. Cre-mediated recombination of a floxed LacZ reporter provided direct evidence that adult small-intestinal crypts contain more than one active multipotent stem cell, and that these cells can be retained in both small-intestinal and colonic crypts for at least 80 d. A method was developed to recover epithelial cells from crypts with or without recombination for subsequent gene expression profiling. Stained sections of intestine were used to create electronic image templates to guide laser capture microdissection (LCM) of adjacent frozen sections. This navigated form of LCM overcomes problems with mRNA degradation encountered when cells are marked directly by immunohistochemical methods. Combining Cre-engineered genetic mosaic mice with navigated-LCM will allow biology and pathobiology to be explored at the junction between normal and perturbed cellular cohorts. PMID- 11050180 TI - A method for fine mapping quantitative trait loci in outbred animal stocks. AB - High-resolution mapping of quantitative trait loci (QTL) in animals has proved to be difficult because the large effect sizes detected in crosses between inbred strains are often caused by numerous linked QTLs, each of small effect. In a study of fearfulness in mice, we have shown it is possible to fine map small effect QTLs in a genetically heterogeneous stock (HS). This strategy is a powerful general method of fine mapping QTLs, provided QTLs detected in crosses between inbred strains that formed the HS can be reliably detected in the HS. We show here that single-marker association analysis identifies only two of five QTLs expected to be segregating in the HS and apparently limits the strategy's usefulness for fine mapping. We solve this problem with a multipoint analysis that assigns the probability that an allele descends from each progenitor in the HS. The analysis does not use pedigrees but instead requires information about the HS founder haplotypes. With this method we mapped all three previously undetected loci [chromosome (Chr.) 1 logP 4.9, Chr. 10 logP 6.0, Chr. 15 logP 4.0]. We show that the reason for the failure of single-marker association to detect QTLs is its inability to distinguish opposing phenotypic effects when they occur on the same marker allele. We have developed a robust method of fine mapping QTLs in genetically heterogeneous animals and suggest it is now cost effective to undertake genomewide high-resolution analysis of complex traits in parallel on the same set of mice. PMID- 11050179 TI - Endogenous regulators of G protein signaling proteins regulate presynaptic inhibition at rat hippocampal synapses. AB - Presynaptic inhibition mediated by G protein-coupled receptors (GPCRs) can develop and decay in a few seconds. This time course is too rapid to be accounted for by the intrinsic GTPase activity of Galpha subunits alone. Here, we test the hypothesis that endogenous regulators of G protein signaling (RGS proteins) are required for rapid, brief presynaptic inhibition. Endogenous G protein alpha subunits were uncoupled from GPCRs by treating cultures with pertussis toxin (PTX). Adenoviral expression of mutant PTX-insensitive (PTX-i) Galpha(i1-3) or Galpha(o) subunits rescued adenosine-induced presynaptic inhibition in cultured hippocampal neurons. Expression of double mutant Galpha(i1) or Galpha(o) subunits that were both PTX-insensitive and unable to bind RGS proteins (PTX/RGS-i) also rescued presynaptic inhibition. Presynaptic inhibition mediated by PTX/RGS-i subunits decayed much more slowly after agonist removal than that mediated by PTX i subunits or native G proteins. The onset of presynaptic inhibition mediated by PTX/RGS-i Galpha(o) was also slower than that mediated by PTX-i Galpha(o). In contrast, the onset of presynaptic inhibition mediated by PTX/RGS-i Galpha(i1) was similar to that mediated by PTX-i Galpha(i1). These results suggest that endogenous RGS proteins regulate the time course of G protein signaling in mammalian central nervous system presynaptic terminals. PMID- 11050181 TI - Geophysical, archaeological, and historical evidence support a solar-output model for climate change. AB - Although the processes of climate change are not completely understood, an important causal candidate is variation in total solar output. Reported cycles in various climate-proxy data show a tendency to emulate a fundamental harmonic sequence of a basic solar-cycle length (11 years) multiplied by 2(N) (where N equals a positive or negative integer). A simple additive model for total solar output variations was developed by superimposing a progression of fundamental harmonic cycles with slightly increasing amplitudes. The timeline of the model was calibrated to the Pleistocene/Holocene boundary at 9,000 years before present. The calibrated model was compared with geophysical, archaeological, and historical evidence of warm or cold climates during the Holocene. The evidence of periods of several centuries of cooler climates worldwide called "little ice ages," similar to the period anno Domini (A.D.) 1280-1860 and reoccurring approximately every 1,300 years, corresponds well with fluctuations in modeled solar output. A more detailed examination of the climate sensitive history of the last 1, 000 years further supports the model. Extrapolation of the model into the future suggests a gradual cooling during the next few centuries with intermittent minor warmups and a return to near little-ice-age conditions within the next 500 years. This cool period then may be followed approximately 1,500 years from now by a return to altithermal conditions similar to the previous Holocene Maximum. PMID- 11050182 TI - Purification, molecular cloning, and sequence analysis of sucrose-6F-phosphate phosphohydrolase from plants. AB - Sucrose-6(F)-phosphate phosphohydrolase (SPP; EC ) catalyzes the final step in the pathway of sucrose biosynthesis and is the only enzyme of photosynthetic carbon assimilation for which the gene has not been identified. The enzyme was purified to homogeneity from rice (Oryza sativa L.) leaves and partially sequenced. The rice leaf enzyme is a dimer with a native molecular mass of 100 kDa and a subunit molecular mass of 50 kDa. The enzyme is highly specific for sucrose 6(F)-phosphate with a K(m) of 65 microM and a specific activity of 1250 micromol min(-1) mg(-1) protein. The activity is dependent on Mg(2+) with a remarkably low K(a) of 8-9 microM and is weakly inhibited by sucrose. Three peptides from cleavage of the purified rice SPP with endoproteinase Lys-C showed similarity to the deduced amino acid sequences of three predicted open reading frames (ORF) in the Arabidopsis thaliana genome and one in the genome of the cyanobacterium Synechocystis sp. PCC6803, as well as cDNA clones from Arabidopsis, maize, and other species in the GenBank database of expressed sequence tags. The putative maize SPP cDNA clone contained an ORF encoding a 420 amino acid polypeptide. Heterologous expression in Escherichia coli showed that this cDNA clone encoded a functional SPP enzyme. The 260-amino acid N-terminal catalytic domain of the maize SPP is homologous to the C-terminal region of sucrose-phosphate synthase. A PSI-BLAST search of the GenBank database indicated that the maize SPP is a member of the haloacid dehalogenase hydrolase/phosphatase superfamily. PMID- 11050183 TI - A targeted mutation in the IL-4Ralpha gene protects mice against autoimmune diabetes. AB - Autoimmune insulin-dependent diabetes mellitus (IDDM) occurs spontaneously in mice-bearing transgenes encoding the influenza hemagglutinin under the control of the rat insulin promoter and a T cell receptor specific for an hemagglutinin peptide associated with I-E(d). Such "double transgenic" mice expressing wild type or targeted IL-4Ralpha genes were examined for the onset of IDDM. Eight of 11 mice homozygous for wild-type IL-4Ralpha were hyperglycemic by 8 weeks of age, whereas only 1 of 16 mice homozygous for the targeted allele were hyperglycemic at this time. Most 1L-4Ralpha-/- mice remained normoglycemic to 36 weeks of age. Although only 10% of double transgenic mice homozygous for the wild-type IL 4Ralpha allele survived to 30 weeks, 80% of mice homozygous for the targeted allele did so. Heterozygous mice displayed an intermediate frequency of diabetes. Even as late as 270 days of age, mice homozygous for the targeted allele had no insulitis or only peri-insulitis. Thus, the inability to respond to IL-4 and/or IL-13 protects mice against IDDM in this model of autoimmunity. PMID- 11050184 TI - Remarkable statistical behavior for truncated Burgers-Hopf dynamics. AB - A simplified one-dimensional model system is introduced and studied here that exhibits intrinsic chaos with many degrees of freedom as well as increased predictability and slower decay of correlations for the large-scale features of the system. These are important features in common with vastly more complex problems involving climate modeling or molecular biological systems. This model is a suitable approximation of the Burgers-Hopf equation involving Galerkin projection on Fourier modes. The model has a detailed mathematical structure that leads to a well-defined equilibrium statistical theory as well as a simple scaling theory for correlations. The numerical evidence presented here strongly supports the behavior predicted from these statistical theories. Unlike the celebrated dissipative and dispersive approximations of the Burgers-Hopf equation, which exhibit exactly solvable and/or completely integrable behavior, these model approximations have strong intrinsic chaos with ergodic behavior. PMID- 11050185 TI - Phosphorylation near nuclear localization signal regulates nuclear import of adenomatous polyposis coli protein. AB - Mutation of the adenomatous polyposis coli (APC) gene is an early step in the development of colorectal carcinomas. APC protein is located in both the cytoplasm and the nucleus. The objective of this study was to define the nuclear localization signals (NLSs) in APC protein. APC contains two potential NLSs comprising amino acids 1767-1772 (NLS1(APC)) and 2048-2053 (NLS2(APC)). Both APC NLSs are well conserved among human, mouse, rat, and fly. NLS1(APC) and NLS2(APC) each were sufficient to target the cytoplasmic protein beta-galactosidase to the nucleus. Mutational analysis of APC demonstrated that both NLSs were necessary for optimal nuclear import of full-length APC protein. Alignment of NLS2(APC) with the simian virus 40 large T antigen NLS (NLS(SV40 T-ag)) revealed sequence similarity extending to adjacent phosphorylation sites. Changing a serine residue (Ser(2054)) to aspartic acid mutated the potential protein kinase A site adjacent to NLS2(APC), resulting in both inhibition of the NLS2(APC)-mediated nuclear import of a chimeric beta-galactosidase fusion protein and a reduction of full length APC nuclear localization. Our data provide evidence that control of APC's nuclear import through phosphorylation is a potential mechanism for regulating APC's nuclear activity. PMID- 11050186 TI - Cooperative subunit interactions within the oligomeric envelope glycoprotein of HIV-1: functional complementation of specific defects in gp120 and gp41. AB - The envelope glycoprotein (Env) of HIV-1 is displayed on the surface of the virion or infected cell as an oligomer of multiple gp120/gp41 complexes. We sought to unravel the relationships between this oligomeric structure and the requirements for sequential interactions with CD4 and coreceptor (CCR5 or CXCR4). We used a quantitative cell fusion assay to examine the effects of coexpressing pairs of Envs, each nonfunctional because of a specific defect in one of the essential properties. We observed efficient fusion activity upon coexpression of two Env variants, one containing a gp41 subunit with a mutated fusion peptide and the other containing a gp120 subunit with a mutated CD4 binding site or a mismatched coreceptor specificity. We also observed fusion upon coexpression of two Env variants with distinct gp120 defects, i.e., a CD4 binding site mutation and the incorrect coreceptor specificity determinants. Coimmunoprecipitation experiments verified the efficient formation of mixed oligomers, suggesting that the observed fusion reflected subunit complementation within the oligomeric complex. These results support a model in which cooperative subunit interactions within the Env oligomer result in concerted conformational changes upon receptor binding, resulting in activation for fusion. The implications of these findings for Env function and virus neutralization are discussed. PMID- 11050187 TI - "Kiss and run" exocytosis at hippocampal synapses. AB - We have combined electrophysiology and imaging to measure the release of neurotransmitter and fluorescent dye at synapses of cultured hippocampal neurons. These experiments have revealed a "kiss and run" mode of exocytosis in which synaptic vesicles release glutamate normally but do not permit dye to enter or escape from the vesicle. During "kiss and run," the vesicle interior may be exposed very transiently (<6 ms), or a special configuration of the fusion pore may prevent dye exchange. We estimate that about 20% of the vesicles normally use this "kiss and run" pathway, and that the fraction of "kiss and run" events can be increased to over 80% by superfusing the synapses with hypertonic solution. PMID- 11050189 TI - The transition from quantity to quality: a neglected causal mechanism in accounting for social evolution. AB - Students of social evolution are concerned not only with the general course it has followed, but also with the mechanisms that have brought it about. One such mechanism comes into play when the quantitative increase in some entity, usually population, reaching a certain threshold, gives rise to a qualitative change in the structure of a society. This mechanism, first recognized by Hegel, was seized on by Marx and Engels. However, neither they nor their current followers among anthropologists have made much use of it in attempting to explain social evolution. But as this paper attempts to show, in those few instances when the mechanism has been invoked, it has heightened our understanding of the process of social evolution. And, it is argued, if the mechanism were more widely applied, further understanding of the course of evolution could be expected to result. PMID- 11050188 TI - A unique loop in the DNA-binding crevice of bacteriophage T7 DNA polymerase influences primer utilization. AB - The three-dimensional structure of bacteriophage T7 DNA polymerase reveals the presence of a loop of 4 aa (residues 401-404) within the DNA-binding groove; this loop is not present in other members of the DNA polymerase I family. A genetically altered T7 DNA polymerase, T7 polDelta401-404, lacking these residues, has been characterized biochemically. The polymerase activity of T7 polDelta401-404 on primed M13 single-stranded DNA template is one-third of the wild-type enzyme and has a 3'-to-5' exonuclease activity indistinguishable from that of wild-type T7 DNA polymerase. T7 polDelta401-404 polymerizes nucleotides processively on a primed M13 single-stranded DNA template. T7 DNA polymerase cannot initiate de novo DNA synthesis; it requires tetraribonucleotides synthesized by the primase activity of the T7 gene 4 protein to serve as primers. T7 primase-dependent DNA synthesis on single-stranded DNA is 3- to 6-fold less with T7 polDelta401-404 compared with the wild-type enzyme. Furthermore, the altered polymerase is defective (10-fold) in its ability to use preformed tetraribonucleotides to initiate DNA synthesis in the presence of gene 4 protein. The location of the loop places it in precisely the position to interact with the tetraribonucleotide primer and, presumably, with the T7 gene 4 primase. Gene 4 protein also provides helicase activity for the replication of duplex DNA. T7 polDelta401-404 and T7 gene 4 protein catalyze strand-displacement DNA synthesis at nearly the same rate as does wild-type polymerase and T7 gene 4 protein, suggesting that the coupling of helicase and polymerase activities is unaffected. PMID- 11050190 TI - From mouse to human: fine mapping of quantitative trait loci in a model organism. PMID- 11050191 TI - The self-same beat of time's wide wings. PMID- 11050192 TI - Can allosteric regulation be predicted from structure? PMID- 11050193 TI - Channel structure and drug-induced cardiac arrhythmias. PMID- 11050194 TI - Facilitating structural transitions in DNA. PMID- 11050195 TI - Ecological mechanisms of extinction. PMID- 11050196 TI - Auditory neuroscience: development, transduction, and integration. AB - Hearing underlies our ability to locate sound sources in the environment, our appreciation of music, and our ability to communicate. Participants in the National Academy of Sciences colloquium on Auditory Neuroscience: Development, Transduction, and Integration presented research results bearing on four key issues in auditory research. How does the complex inner ear develop? How does the cochlea transduce sounds into electrical signals? How does the brain's ability to compute the location of a sound source develop? How does the forebrain analyze complex sounds, particularly species-specific communications? This article provides an introduction to the papers stemming from the meeting. PMID- 11050198 TI - Molecular genetics of pattern formation in the inner ear: do compartment boundaries play a role? AB - The membranous labyrinth of the inner ear establishes a precise geometrical topology so that it may subserve the functions of hearing and balance. How this geometry arises from a simple ectodermal placode is under active investigation. The placode invaginates to form the otic cup, which deepens before pinching off to form the otic vesicle. By the vesicle stage many genes expressed in the developing ear have assumed broad, asymmetrical expression domains. We have been exploring the possibility that these domains may reflect developmental compartments that are instrumental in specifying the location and identity of different parts of the ear. The boundaries between compartments are proposed to be the site of inductive interactions required for this specification. Our work has shown that sensory organs and the endolymphatic duct each arise near the boundaries of broader gene expression domains, lending support to this idea. A further prediction of the model, that the compartment boundaries will also represent lineage-restriction compartments, is supported in part by fate mapping the otic cup. Our data suggest that two lineage-restriction boundaries intersect at the dorsal pole of the otocyst, a convergence that may be critical for the specification of endolymphatic duct outgrowth. We speculate that the patterning information necessary to establish these two orthogonal boundaries may emanate, in part, from the hindbrain. The compartment boundary model of ear development now needs to be tested through a variety of experimental perturbations, such as the removal of boundaries, the generation of ectopic boundaries, and/or changes in compartment identity. PMID- 11050197 TI - Notch signaling in the development of the inner ear: lessons from Drosophila. AB - The sensory patches in the ear of a vertebrate can be compared with the mechanosensory bristles of a fly. This comparison has led to the discovery that lateral inhibition mediated by the Notch cell-cell signaling pathway, first characterized in Drosophila and crucial for bristle development, also has a key role in controlling the pattern of sensory hair cells and supporting cells in the ear. We review the arguments for considering the sensory patches of the vertebrate ear and bristles of the insect to be homologous structures, evolved from a common ancestral mechanosensory organ, and we examine more closely the role of Notch signaling in each system. Using viral vectors to misexpress components of the Notch pathway in the chick ear, we show that a simple lateral inhibition model based on feedback regulation of the Notch ligand Delta is inadequate for the ear just as it is for the fly bristle. The Notch ligand Serrate1, expressed in supporting cells in the ear, is regulated by lateral induction, not lateral inhibition; commitment to become a hair cell is not simply controlled by levels of expression of the Notch ligands Delta1, Serrate1, and Serrate2 in the neighbors of the nascent hair cell; and at least one factor, Numb, capable of blocking reception of lateral inhibition is concentrated in hair cells. These findings reinforce the parallels between the vertebrate ear and the fly bristle and show how study of the insect system can help us understand the vertebrate. PMID- 11050199 TI - Patterning of the mammalian cochlea. AB - The mammalian cochlea is sophisticated in its function and highly organized in its structure. Although the anatomy of this sense organ has been well documented, the molecular mechanisms underlying its development have remained elusive. Information generated from mutant and knockout mice in recent years has increased our understanding of cochlear development and physiology. This article discusses factors important for the development of the inner ear and summarizes cochlear phenotypes of mutant and knockout mice, particularly Otx and Otx2. We also present data on gross development of the mouse cochlea. PMID- 11050202 TI - Two mechanisms for transducer adaptation in vertebrate hair cells. AB - Deflection of the hair bundle atop a sensory hair cell modulates the open probability of mechanosensitive ion channels. In response to sustained deflections, hair cells adapt. Two fundamentally distinct models have been proposed to explain transducer adaptation. Both models support the notion that channel open probability is modulated by calcium that enters via the transduction channels. Both also suggest that the primary effect of adaptation is to shift the deflection-response [I(X)] relationship in the direction of the applied stimulus, thus maintaining hair bundle sensitivity. The models differ in several respects. They operate on different time scales: the faster on the order of a few milliseconds or less and the slower on the order of 10 ms or more. The model proposed to explain fast adaptation suggests that calcium enters and binds at or near the transduction channels to stabilize a closed conformation. The model proposed to explain the slower adaptation suggests that adaptation is mediated by an active, force-generating process that regulates the effective stimulus applied to the transduction channels. Here we discuss the evidence in support of each model and consider the possibility that both may function to varying degrees in hair cells of different species and sensory organs. PMID- 11050201 TI - Hair cell recovery in mitotically blocked cultures of the bullfrog saccule. AB - Hair cells in many nonmammalian vertebrates are regenerated by the mitotic division of supporting cell progenitors and the differentiation of the resulting progeny into new hair cells and supporting cells. Recent studies have shown that nonmitotic hair cell recovery after aminoglycoside-induced damage can also occur in the vestibular organs. Using hair cell and supporting cell immunocytochemical markers, we have used confocal and electron microscopy to examine the fate of damaged hair cells and the origin of immature hair cells after gentamicin treatment in mitotically blocked cultures of the bullfrog saccule. Extruding and fragmenting hair cells, which undergo apoptotic cell death, are replaced by scar formations. After losing their bundles, sublethally damaged hair cells remain in the sensory epithelium for prolonged periods, acquiring supporting cell-like morphology and immunoreactivity. These modes of damage appear to be mutually exclusive, implying that sublethally damaged hair cells repair their bundles. Transitional cells, coexpressing hair cell and supporting cell markers, are seen near scar formations created by the expansion of neighboring supporting cells. Most of these cells have morphology and immunoreactivity similar to that of sublethally damaged hair cells. Ultrastructural analysis also reveals that most immature hair cells had autophagic vacuoles, implying that they originated from damaged hair cells rather than supporting cells. Some transitional cells are supporting cells participating in scar formations. Supporting cells also decrease in number during hair cell recovery, supporting the conclusion that some supporting cells undergo phenotypic conversion into hair cells without an intervening mitotic event. PMID- 11050203 TI - Cochlear mechanisms from a phylogenetic viewpoint. AB - The hearing organ of the inner ear was the last of the paired sense organs of amniotes to undergo formative evolution. As a mechanical sensory organ, the inner ear hearing organ's function depends highly on its physical structure. Comparative studies suggest that the hearing organ of the earliest amniote vertebrates was small and simple, but possessed hair cells with a cochlear amplifier mechanism, electrical frequency tuning, and incipient micromechanical tuning. The separation of the different groups of amniotes from the stem reptiles occurred relatively early, with the ancestors of the mammals branching off first, approximately 320 million years ago. The evolution of the hearing organ in the three major lines of the descendents of the stem reptiles (e.g., mammals, birds crocodiles, and lizards-snakes) thus occurred independently over long periods of time. Dramatic and parallel improvements in the middle ear initiated papillar elongation in all lineages, accompanied by increased numbers of sensory cells with enhanced micromechanical tuning and group-specific hair-cell specializations that resulted in unique morphological configurations. This review aims not only to compare structure and function across classification boundaries (the comparative approach), but also to assess how and to what extent fundamental mechanisms were influenced by selection pressures in times past (the phylogenetic viewpoint). PMID- 11050200 TI - Cellular studies of auditory hair cell regeneration in birds. AB - A decade ago it was discovered that mature birds are able to regenerate hair cells, the receptors for auditory perception. This surprising finding generated hope in the field of auditory neuroscience that new hair cells someday may be coaxed to form in another class of warm-blooded vertebrates, mammals. We have made considerable progress toward understanding some cellular and molecular events that lead to hair cell regeneration in birds. This review discusses our current understanding of avian hair cell regeneration, with some comparisons to other vertebrate classes and other regenerative systems. PMID- 11050204 TI - Mechanical bases of frequency tuning and neural excitation at the base of the cochlea: comparison of basilar-membrane vibrations and auditory-nerve-fiber responses in chinchilla. AB - We review the mechanical origin of auditory-nerve excitation, focusing on comparisons of the magnitudes and phases of basilar-membrane (BM) vibrations and auditory-nerve fiber responses to tones at a basal site of the chinchilla cochlea with characteristic frequency approximately 9 kHz located 3.5 mm from the oval window. At this location, characteristic frequency thresholds of fibers with high spontaneous activity correspond to magnitudes of BM displacement or velocity in the order of 1 nm or 50 microm/s. Over a wide range of stimulus frequencies, neural thresholds are not determined solely by BM displacement but rather by a function of both displacement and velocity. Near-threshold, auditory-nerve responses to low-frequency tones are synchronous with peak BM velocity toward scala tympani but at 80-90 dB sound pressure level (in decibels relative to 20 microPascals) and at 100-110 dB sound pressure level responses undergo two large phase shifts approaching 180 degrees. These drastic phase changes have no counterparts in BM vibrations. Thus, although at threshold levels the encoding of BM vibrations into spike trains appears to involve only relatively minor signal transformations, the polarity of auditory-nerve responses does not conform with traditional views of how BM vibrations are transmitted to the inner hair cells. The response polarity at threshold levels, as well as the intensity-dependent phase changes, apparently reflect micromechanical interactions between the organ of Corti, the tectorial membrane and the subtectorial fluid, and/or electrical and synaptic processes at the inner hair cells. PMID- 11050205 TI - The spatial and temporal representation of a tone on the guinea pig basilar membrane. AB - In the mammalian cochlea, the basilar membrane's (BM) mechanical responses are amplified, and frequency tuning is sharpened through active feedback from the electromotile outer hair cells (OHCs). To be effective, OHC feedback must be delivered to the correct region of the BM and introduced at the appropriate time in each cycle of BM displacement. To investigate when OHCs contribute to cochlear amplification, a laser-diode interferometer was used to measure tone-evoked BM displacements in the basal turn of the guinea pig cochlea. Measurements were made at multiple sites across the width of the BM, which are tuned to the same characteristic frequency (CF). In response to CF tones, the largest displacements occur in the OHC region and phase lead those measured beneath the outer pillar cells and adjacent to the spiral ligament by about 90 degrees. Postmortem, responses beneath the OHCs are reduced by up to 65 dB, and all regions across the width of the BM move in unison. We suggest that OHCs amplify BM responses to CF tones when the BM is moving at maximum velocity. In regions of the BM where OHCs contribute to its motion, the responses are compressive and nonlinear. We measured the distribution of nonlinear compressive vibrations along the length of the BM in response to a single frequency tone and estimated that OHC amplification is restricted to a 1.25- to 1.40-mm length of BM centered on the CF place. PMID- 11050206 TI - Molecular mechanisms of sound amplification in the mammalian cochlea. AB - Mammalian hearing depends on the enhanced mechanical properties of the basilar membrane within the cochlear duct. The enhancement arises through the action of outer hair cells that act like force generators within the organ of Corti. Simple considerations show that underlying mechanism of somatic motility depends on local area changes within the lateral membrane of the cell. The molecular basis for this phenomenon is a dense array of particles that are inserted into the basolateral membrane and that are capable of sensing membrane potential field. We show here that outer hair cells selectively take up fructose, at rates high enough to suggest that a sugar transporter may be part of the motor complex. The relation of these findings to a recent candidate for the molecular motor is also discussed. PMID- 11050209 TI - Linear and nonlinear pathways of spectral information transmission in the cochlear nucleus. AB - At the level of the cochlear nucleus (CN), the auditory pathway divides into several parallel circuits, each of which provides a different representation of the acoustic signal. Here, the representation of the power spectrum of an acoustic signal is analyzed for two CN principal cells-chopper neurons of the ventral CN and type IV neurons of the dorsal CN. The analysis is based on a weighting function model that relates the discharge rate of a neuron to first- and second-order transformations of the power spectrum. In chopper neurons, the transformation of spectral level into rate is a linear (i.e., first-order) or nearly linear function. This transformation is a predominantly excitatory process involving multiple frequency components, centered in a narrow frequency range about best frequency, that usually are processed independently of each other. In contrast, type IV neurons encode spectral information linearly only near threshold. At higher stimulus levels, these neurons are strongly inhibited by spectral notches, a behavior that cannot be explained by level transformations of first- or second-order. Type IV weighting functions reveal complex excitatory and inhibitory interactions that involve frequency components spanning a wider range than that seen in choppers. These findings suggest that chopper and type IV neurons form parallel pathways of spectral information transmission that are governed by two different mechanisms. Although choppers use a predominantly linear mechanism to transmit tonotopic representations of spectra, type IV neurons use highly nonlinear processes to signal the presence of wide-band spectral features. PMID- 11050207 TI - Putting ion channels to work: mechanoelectrical transduction, adaptation, and amplification by hair cells. AB - As in other excitable cells, the ion channels of sensory receptors produce electrical signals that constitute the cellular response to stimulation. In photoreceptors, olfactory neurons, and some gustatory receptors, these channels essentially report the results of antecedent events in a cascade of chemical reactions. The mechanoelectrical transduction channels of hair cells, by contrast, are coupled directly to the stimulus. As a consequence, the mechanical properties of these channels shape our hearing process from the outset of transduction. Channel gating introduces nonlinearities prominent enough to be measured and even heard. Channels provide a feedback signal that controls the transducer's adaptation to large stimuli. Finally, transduction channels participate in an amplificatory process that sensitizes and sharpens hearing. PMID- 11050208 TI - Detection of synchrony in the activity of auditory nerve fibers by octopus cells of the mammalian cochlear nucleus. AB - The anatomical and biophysical specializations of octopus cells allow them to detect the coincident firing of groups of auditory nerve fibers and to convey the precise timing of that coincidence to their targets. Octopus cells occupy a sharply defined region of the most caudal and dorsal part of the mammalian ventral cochlear nucleus. The dendrites of octopus cells cross the bundle of auditory nerve fibers just proximal to where the fibers leave the ventral and enter the dorsal cochlear nucleus, each octopus cell spanning about one-third of the tonotopic array. Octopus cells are excited by auditory nerve fibers through the activation of rapid, calcium-permeable, alpha-amino-3-hydroxy-5-methyl-4 isoxazole-propionate receptors. Synaptic responses are shaped by the unusual biophysical characteristics of octopus cells. Octopus cells have very low input resistances (about 7 M Omega), and short time constants (about 200 microsec) as a consequence of the activation at rest of a hyperpolarization-activated mixed cation conductance and a low-threshold, depolarization-activated potassium conductance. The low input resistance causes rapid synaptic currents to generate rapid and small synaptic potentials. Summation of small synaptic potentials from many fibers is required to bring an octopus cell to threshold. Not only does the low input resistance make individual excitatory postsynaptic potentials brief so that they must be generated within 1 msec to sum but also the voltage-sensitive conductances of octopus cells prevent firing if the activation of auditory nerve inputs is not sufficiently synchronous and depolarization is not sufficiently rapid. In vivo in cats, octopus cells can fire rapidly and respond with exceptionally well-timed action potentials to periodic, broadband sounds such as clicks. Thus both the anatomical specializations and the biophysical specializations make octopus cells detectors of the coincident firing of their auditory nerve fiber inputs. PMID- 11050210 TI - Cellular mechanisms for resolving phase ambiguity in the owl's inferior colliculus. AB - Both mammals and birds use the interaural time difference (ITD) for localization of sound in the horizontal plane. They may localize either real or phantom sound sources, when the signal consists of a narrow frequency band. This ambiguity does not occur with broadband signals. A plot of impulse rates or amplitude of excitatory postsynaptic potentials against ITDs (ITD curve) consists of peaks and troughs. In the external nucleus (ICX) of the owl's inferior colliculus, ITD curves show multiple peaks when the signal is narrow-band, such as tones. Of these peaks, one occurs at ITDi, which is independent of frequency, and others at ITDi +/- T, where T is the tonal period. The ITD curve of the same neuron shows a large peak (main peak) at ITDi and no or small peaks (side peaks) at ITDi +/- T, when the signal is broadband. ITD curves for postsynaptic potentials indicate that ICX neurons integrate the results of binaural cross-correlation in different frequency bands. However, the difference between the main and side peaks is small. ICX neurons further enhance this difference in the process of converting membrane potentials to impulse rates. Inhibition also appears to augment the difference between the main and side peaks. PMID- 11050211 TI - Subdivisions of auditory cortex and processing streams in primates. AB - The auditory system of monkeys includes a large number of interconnected subcortical nuclei and cortical areas. At subcortical levels, the structural components of the auditory system of monkeys resemble those of nonprimates, but the organization at cortical levels is different. In monkeys, the ventral nucleus of the medial geniculate complex projects in parallel to a core of three primary like auditory areas, AI, R, and RT, constituting the first stage of cortical processing. These areas interconnect and project to the homotopic and other locations in the opposite cerebral hemisphere and to a surrounding array of eight proposed belt areas as a second stage of cortical processing. The belt areas in turn project in overlapping patterns to a lateral parabelt region with at least rostral and caudal subdivisions as a third stage of cortical processing. The divisions of the parabelt distribute to adjoining auditory and multimodal regions of the temporal lobe and to four functionally distinct regions of the frontal lobe. Histochemically, chimpanzees and humans have an auditory core that closely resembles that of monkeys. The challenge for future researchers is to understand how this complex system in monkeys analyzes and utilizes auditory information. PMID- 11050212 TI - Mechanisms and streams for processing of "what" and "where" in auditory cortex. AB - The functional specialization and hierarchical organization of multiple areas in rhesus monkey auditory cortex were examined with various types of complex sounds. Neurons in the lateral belt areas of the superior temporal gyrus were tuned to the best center frequency and bandwidth of band-passed noise bursts. They were also selective for the rate and direction of linear frequency modulated sweeps. Many neurons showed a preference for a limited number of species-specific vocalizations ("monkey calls"). These response selectivities can be explained by nonlinear spectral and temporal integration mechanisms. In a separate series of experiments, monkey calls were presented at different spatial locations, and the tuning of lateral belt neurons to monkey calls and spatial location was determined. Of the three belt areas the anterolateral area shows the highest degree of specificity for monkey calls, whereas neurons in the caudolateral area display the greatest spatial selectivity. We conclude that the cortical auditory system of primates is divided into at least two processing streams, a spatial stream that originates in the caudal part of the superior temporal gyrus and projects to the parietal cortex, and a pattern or object stream originating in the more anterior portions of the lateral belt. A similar division of labor can be seen in human auditory cortex by using functional neuroimaging. PMID- 11050213 TI - The corticofugal system for hearing: recent progress. AB - Peripheral auditory neurons are tuned to single frequencies of sound. In the central auditory system, excitatory (or facilitatory) and inhibitory neural interactions take place at multiple levels and produce neurons with sharp level tolerant frequency-tuning curves, neurons tuned to parameters other than frequency, cochleotopic (frequency) maps, which are different from the peripheral cochleotopic map, and computational maps. The mechanisms to create the response properties of these neurons have been considered to be solely caused by divergent and convergent projections of neurons in the ascending auditory system. The recent research on the corticofugal (descending) auditory system, however, indicates that the corticofugal system adjusts and improves auditory signal processing by modulating neural responses and maps. The corticofugal function consists of at least the following subfunctions. (i) Egocentric selection for short-term modulation of auditory signal processing according to auditory experience. Egocentric selection, based on focused positive feedback associated with widespread lateral inhibition, is mediated by the cortical neural net working together with the corticofugal system. (ii) Reorganization for long-term modulation of the processing of behaviorally relevant auditory signals. Reorganization is based on egocentric selection working together with nonauditory systems. (iii) Gain control based on overall excitatory, facilitatory, or inhibitory corticofugal modulation. Egocentric selection can be viewed as selective gain control. (iv) Shaping (or even creation) of response properties of neurons. Filter properties of neurons in the frequency, amplitude, time, and spatial domains can be sharpened by the corticofugal system. Sharpening of tuning is one of the functions of egocentric selection. PMID- 11050214 TI - Traces of learning in the auditory localization pathway. AB - One of the fascinating properties of the central nervous system is its ability to learn: the ability to alter its functional properties adaptively as a consequence of the interactions of an animal with the environment. The auditory localization pathway provides an opportunity to observe such adaptive changes and to study the cellular mechanisms that underlie them. The midbrain localization pathway creates a multimodal map of space that represents the nervous system's associations of auditory cues with locations in visual space. Various manipulations of auditory or visual experience, especially during early life, that change the relationship between auditory cues and locations in space lead to adaptive changes in auditory localization behavior and to corresponding changes in the functional and anatomical properties of this pathway. Traces of this early learning persist into adulthood, enabling adults to reacquire patterns of connectivity that were learned initially during the juvenile period. PMID- 11050215 TI - Plasticity in the neural coding of auditory space in the mammalian brain. AB - Sound localization relies on the neural processing of monaural and binaural spatial cues that arise from the way sounds interact with the head and external ears. Neurophysiological studies of animals raised with abnormal sensory inputs show that the map of auditory space in the superior colliculus is shaped during development by both auditory and visual experience. An example of this plasticity is provided by monaural occlusion during infancy, which leads to compensatory changes in auditory spatial tuning that tend to preserve the alignment between the neural representations of visual and auditory space. Adaptive changes also take place in sound localization behavior, as demonstrated by the fact that ferrets raised and tested with one ear plugged learn to localize as accurately as control animals. In both cases, these adjustments may involve greater use of monaural spectral cues provided by the other ear. Although plasticity in the auditory space map seems to be restricted to development, adult ferrets show some recovery of sound localization behavior after long-term monaural occlusion. The capacity for behavioral adaptation is, however, task dependent, because auditory spatial acuity and binaural unmasking (a measure of the spatial contribution to the "cocktail party effect") are permanently impaired by chronically plugging one ear, both in infancy but especially in adulthood. Experience-induced plasticity allows the neural circuitry underlying sound localization to be customized to individual characteristics, such as the size and shape of the head and ears, and to compensate for natural conductive hearing losses, including those associated with middle ear disease in infancy. PMID- 11050216 TI - Spatial processing in the auditory cortex of the macaque monkey. AB - The patterns of cortico-cortical and cortico-thalamic connections of auditory cortical areas in the rhesus monkey have led to the hypothesis that acoustic information is processed in series and in parallel in the primate auditory cortex. Recent physiological experiments in the behaving monkey indicate that the response properties of neurons in different cortical areas are both functionally distinct from each other, which is indicative of parallel processing, and functionally similar to each other, which is indicative of serial processing. Thus, auditory cortical processing may be similar to the serial and parallel "what" and "where" processing by the primate visual cortex. If "where" information is serially processed in the primate auditory cortex, neurons in cortical areas along this pathway should have progressively better spatial tuning properties. This prediction is supported by recent experiments that have shown that neurons in the caudomedial field have better spatial tuning properties than neurons in the primary auditory cortex. Neurons in the caudomedial field are also better than primary auditory cortex neurons at predicting the sound localization ability across different stimulus frequencies and bandwidths in both azimuth and elevation. These data support the hypothesis that the primate auditory cortex processes acoustic information in a serial and parallel manner and suggest that this may be a general cortical mechanism for sensory perception. PMID- 11050217 TI - Song selectivity and sensorimotor signals in vocal learning and production. AB - Bird song, like human speech, is a learned vocal behavior that requires auditory feedback. Both as juveniles, while they learn to sing, and as adults, songbirds use auditory feedback to compare their own vocalizations with an internal model of a target song. Here we describe experiments that explore a role for the songbird anterior forebrain pathway (AFP), a basal ganglia-forebrain circuit, in evaluating song feedback and modifying vocal output. First, neural recordings in anesthetized, juvenile birds show that single AFP neurons are specialized to process the song stimuli that are compared during sensorimotor learning. AFP neurons are tuned to both the bird's own song and the tutor song, even when these stimuli are manipulated to be very different from each other. Second, behavioral experiments in adult birds demonstrate that lesions to the AFP block the deterioration of song that normally follows deafening. This observation suggests that deafening results in an instructive signal, indicating a mismatch between feedback and the internal song model, and that the AFP is involved in generating or transmitting this instructive signal. Finally, neural recordings from behaving birds reveal robust singing-related activity in the AFP. This activity is likely to originate from premotor areas and could be modulated by auditory feedback of the bird's own voice. One possibility is that this activity represents an efference copy, predicting the sensory consequences of motor commands. Overall, these studies illustrate that sensory and motor processes are highly interrelated in this circuit devoted to vocal learning, as is true for brain areas involved in speech. PMID- 11050218 TI - On cortical coding of vocal communication sounds in primates. AB - Understanding how the brain processes vocal communication sounds is one of the most challenging problems in neuroscience. Our understanding of how the cortex accomplishes this unique task should greatly facilitate our understanding of cortical mechanisms in general. Perception of species-specific communication sounds is an important aspect of the auditory behavior of many animal species and is crucial for their social interactions, reproductive success, and survival. The principles of neural representations of these behaviorally important sounds in the cerebral cortex have direct implications for the neural mechanisms underlying human speech perception. Our progress in this area has been relatively slow, compared with our understanding of other auditory functions such as echolocation and sound localization. This article discusses previous and current studies in this field, with emphasis on nonhuman primates, and proposes a conceptual platform to further our exploration of this frontier. It is argued that the prerequisite condition for understanding cortical mechanisms underlying communication sound perception and production is an appropriate animal model. Three issues are central to this work: (i) neural encoding of statistical structure of communication sounds, (ii) the role of behavioral relevance in shaping cortical representations, and (iii) sensory-motor interactions between vocal production and perception systems. PMID- 11050220 TI - Attributes of an alluvial river and their relation to water policy and management. AB - Rivers around the world are being regulated by dams to accommodate the needs of a rapidly growing global population. These regulatory efforts usually oppose the natural tendency of rivers to flood, move sediment, and migrate. Although an economic benefit, river regulation has come at unforeseen and unevaluated cumulative ecological costs. Historic and contemporary approaches to remedy environmental losses have largely ignored hydrologic, geomorphic, and biotic processes that form and maintain healthy alluvial river ecosystems. Several commonly known concepts that govern how alluvial channels work have been compiled into a set of "attributes" for alluvial river integrity. These attributes provide a minimum checklist of critical geomorphic and ecological processes derived from field observation and experimentation, a set of hypotheses to chart and evaluate strategies for restoring and preserving alluvial river ecosystems. They can guide how to (i) restore alluvial processes below an existing dam without necessarily resorting to extreme measures such as demolishing one, and (ii) preserve alluvial river integrity below proposed dams. Once altered by dam construction, a regulated alluvial river will never function as before. But a scaled-down morphology could retain much of a river's original integrity if key processes addressed in the attributes are explicitly provided. Although such a restoration strategy is an experiment, it may be the most practical solution for recovering regulated alluvial river ecosystems and the species that inhabit them. Preservation or restoration of the alluvial river attributes is a logical policy direction for river management in the future. PMID- 11050219 TI - A new view of language acquisition. AB - At the forefront of debates on language are new data demonstrating infants' early acquisition of information about their native language. The data show that infants perceptually "map" critical aspects of ambient language in the first year of life before they can speak. Statistical properties of speech are picked up through exposure to ambient language. Moreover, linguistic experience alters infants' perception of speech, warping perception in the service of language. Infants' strategies are unexpected and unpredicted by historical views. A new theoretical position has emerged, and six postulates of this position are described. PMID- 11050222 TI - Bounds for cell entries in contingency tables given marginal totals and decomposable graphs. AB - Upper and lower bounds on cell counts in cross-classifications of nonnegative counts play important roles in a number of practical problems, including statistical disclosure limitation, computer tomography, mass transportation, cell suppression, and data swapping. Some features of the Frechet bounds are well known, intuitive, and regularly used by those working on disclosure limitation methods, especially those for two-dimensional tables. We previously have described a series of results relating these bounds to theory on loglinear models for cross-classified counts. This paper provides the actual theory and proofs for the special case of decomposable loglinear models and their related independence graphs. It also includes an extension linked to the structure of reducible graphs and a discussion of the relevance of other results linked to nongraphical loglinear models. PMID- 11050221 TI - Neuromelanin biosynthesis is driven by excess cytosolic catecholamines not accumulated by synaptic vesicles. AB - Melanin, the pigment in hair, skin, eyes, and feathers, protects external tissue from damage by UV light. In contrast, neuromelanin (NM) is found in deep brain regions, specifically in loci that degenerate in Parkinson's disease. Although this distribution suggests a role for NM in Parkinson's disease neurodegeneration, the biosynthesis and function of NM have eluded characterization because of lack of an experimental system. We induced NM in rat substantia nigra and PC12 cell cultures by exposure to l-dihydroxyphenylalanine, which is rapidly converted to dopamine (DA) in the cytosol. This pigment was identical to human NM as assessed by paramagnetic resonance and was localized in double membrane autophagic vacuoles identical to NM granules of human substantia nigra. NM synthesis was abolished by adenoviral-mediated overexpression of the synaptic vesicle catecholamine transporter VMAT2, which decreases cytosolic DA by increasing vesicular accumulation of neurotransmitter. The NM is in a stable complex with ferric iron, and NM synthesis was inhibited by the iron chelator desferrioxamine, indicating that cytosolic DA and dihydroxyphenylalanine are oxidized by iron-mediated catalysis to membrane-impermeant quinones and semiquinones. NM synthesis thus results from excess cytosolic catecholamines not accumulated into synaptic vesicles. The permanent accumulation of excess catechols, quinones, and catechol adducts into a membrane-impermeant substance trapped in organelles may provide an antioxidant mechanism for catecholamine neurons. However, NM in organelles associated with secretory pathways may interfere with signaling, as it delays stimulated neurite outgrowth in PC12 cells. PMID- 11050223 TI - How important are entropic contributions to enzyme catalysis? AB - The idea that enzymes accelerate their reactions by entropic effects has played a major role in many prominent proposals about the origin of enzyme catalysis. This idea implies that the binding to an enzyme active site freezes the motion of the reacting fragments and eliminates their entropic contributions, (delta S(cat)(double dagger))', to the activation energy. It is also implied that the binding entropy is equal to the activation entropy, (delta S(w)(double dagger))', of the corresponding solution reaction. It is, however, difficult to examine this idea by experimental approaches. The present paper defines the entropic proposal in a rigorous way and develops a computer simulation approach that determines (delta S(double dagger))'. This approach allows us to evaluate the differences between (delta S(double dagger))' of an enzymatic reaction and of the corresponding reference reaction in solution. Our approach is used in a study of the entropic contribution to the catalytic reaction of subtilisin. It is found that this contribution is much smaller than previously thought. This result is due to the following: (i) Many of the motions that are free in the reactants state of the reference solution reaction are also free at the transition state. (ii) The binding to the enzyme does not completely freeze the motion of the reacting fragments so that (delta S(double dagger))' in the enzymes is not zero. (iii) The binding entropy is not necessarily equal to (delta S(w)(double dagger))'. PMID- 11050224 TI - Furin-mediated processing in the early secretory pathway: sequential cleavage and degradation of misfolded insulin receptors. AB - Improperly folded membrane proteins are retained in the endoplasmic reticulum and then diverted to a degradative pathway by a network of molecular chaperones and intracellular proteases. Here we report that mutant insulin proreceptors (Pro(62)) retained in the early secretory pathway undergo proteolytic cleavage at a tetrabasic concensus site for the subtilisin-like protease furin (SPC 1), generating two unstable proteolytic intermediates of 80/120 kDa corresponding to alpha (135 kDa) and beta (90 kDa) subunits. These are degraded more rapidly than the uncleaved proreceptor protein. Site-directed mutagenesis of the normal RKRR processing site prevented cleavage. Use of inhibitors and furin-deficient cell lines confirmed that furin is responsible for proreceptor cleavage; furin overexpression increased the degradation of mutant but not wild-type receptors. Together, these results suggest that processing and degradation occur sequentially for mutant proreceptors. PMID- 11050225 TI - A census of glutamine/asparagine-rich regions: implications for their conserved function and the prediction of novel prions. AB - Glutamine/asparagine (Q/N)-rich domains have a high propensity to form self propagating amyloid fibrils. This phenomenon underlies both prion-based inheritance in yeast and aggregation of a number of proteins involved in human neurodegenerative diseases. To examine the prevalence of this phenomenon, complete proteomic sequences of 31 organisms and several incomplete proteomic sequences were examined for Q/N-rich regions. We found that Q/N-rich regions are essentially absent from the thermophilic bacterial and archaeal proteomes. Moreover, the average Q/N content of the proteins in these organisms is markedly lower than in mesophilic bacteria and eukaryotes. Mesophilic bacterial proteomes contain a small number (0-4) of proteins with Q/N-rich regions. Remarkably, Q/N rich domains are found in a much larger number of eukaryotic proteins (107-472 per proteome) with diverse biochemical functions. Analyses of these regions argue they have been evolutionarily selected perhaps as modular "polar zipper" protein protein interaction domains. These data also provide a large pool of potential novel prion-forming proteins, two of which have recently been shown to behave as prions in yeast, thus suggesting that aggregation or prion-like regulation of protein function may be a normal regulatory process for many eukaryotic proteins with a wide variety of functions. PMID- 11050226 TI - Miniaturized metalloproteins: application to iron-sulfur proteins. AB - The miniaturization process applied to rubredoxins generated a class of peptide based metalloprotein models, named METP (miniaturized electron transfer protein). The crystal structure of Desulfovibrio vulgaris rubredoxin was selected as a template for the construction of a tetrahedral (S(gamma)-Cys)(4) iron-binding site. Analysis of the structure showed that a sphere of 17 A in diameter, centered on the metal, circumscribes two unconnected approximately C(2) symmetry related beta-hairpins, each containing the -Cys-(Aaa)(2)-Cys- sequence. These observations provided a starting point for the design of an undecapeptide, which self assembles in the presence of tetrahedrally coordinating metal ions. The METP peptide was synthesized in good yield by standard methodologies. Successful assembly of the METP peptide with Co(II), Zn(II), Fe(II/III), in the expected 2:1 stoichiometry, was proven by UV-visible and circular dichroism spectroscopies. UV visible analysis of the metal complexes indicated the four Cys ligands tetrahedrally arrange around the metal ion, as designed. Circular dichroism measurements on both the free and metal-bound forms revealed that the metal coordination drives the peptide chain to fold into a turned conformation. NMR characterization of the Zn(II)-METP complex fully supported the structure of the designed model. These results prove that METP reproduces the main features of rubredoxin. PMID- 11050227 TI - Molecular basis for modulation of biological function by alternate splicing of the Wilms' tumor suppressor protein. AB - Alternate splicing, leading to the insertion of the tripeptide KTS in the linker between the third and fourth C(2)H(2) zinc fingers, changes both the DNA-binding function and the subnuclear localization of the Wilms' tumor suppressor protein (WT1). We have used NMR relaxation experiments to determine the molecular basis for the differing DNA recognition properties of the WT1-KTS and WT1+KTS isoforms. Our results show that the KTS insertion increases the flexibility of the linker between fingers 3 and 4 and abrogates binding of the fourth zinc finger to its cognate site in the DNA major groove. This represents a mechanism whereby a single zinc-finger gene can be used, through alternate splicing, to fulfill different functions in the cell. PMID- 11050228 TI - A single amino acid substitution (F363I) converts the regiochemistry of the spearmint (-)-limonene hydroxylase from a C6- to a C3-hydroxylase. AB - The essential oils of peppermint and spearmint are distinguished by the position of oxygenation on the constituent p-menthane monoterpenes. Peppermint produces monoterpenes bearing an oxygen at C3, whereas spearmint produces monoterpenes bearing an oxygen at C6. Branching of the monoterpene biosynthetic pathways in these species is determined by two distinct cytochrome P450s that catalyze the regiospecific hydroxylation of (-)-4S-limonene at C3 or C6 exclusively. cDNAs encoding the limonene-3-hydroxylase from peppermint and the limonene-6 hydroxylase from spearmint have been isolated, shown to be 70% identical at the amino acid level, and functionally expressed. A combination of domain swapping and reciprocal site-directed mutagenesis between these two enzymes demonstrated that the exchange of a single residue (F363I) in the spearmint limonene-6 hydroxylase led to complete conversion to the regiospecificity and catalytic efficiency of the peppermint limonene-3-hydroxylase. PMID- 11050229 TI - Biochemistry, mutagenesis, and oligomerization of DsRed, a red fluorescent protein from coral. AB - DsRed is a recently cloned 28-kDa fluorescent protein responsible for the red coloration around the oral disk of a coral of the Discosoma genus. DsRed has attracted tremendous interest as a potential expression tracer and fusion partner that would be complementary to the homologous green fluorescent protein from Aequorea, but very little is known of the biochemistry of DsRed. We now show that DsRed has a much higher extinction coefficient and quantum yield than previously reported, plus excellent resistance to pH extremes and photobleaching. In addition, its 583-nm emission maximum can be further shifted to 602 nm by mutation of Lys-83 to Met. However, DsRed has major drawbacks, such as strong oligomerization and slow maturation. Analytical ultracentrifugation proves DsRed to be an obligate tetramer in vitro, and fluorescence resonance energy transfer measurements and yeast two-hybrid assays verify oligomerization in live cells. Also, DsRed takes days to ripen fully from green to red in vitro or in vivo, and mutations such as Lys-83 to Arg prevent the color change. Many potential cell biological applications of DsRed will require suppression of the tetramerization and acceleration of the maturation. PMID- 11050230 TI - The structure of the chromophore within DsRed, a red fluorescent protein from coral. AB - DsRed, a brilliantly red fluorescent protein, was recently cloned from Discosoma coral by homology to the green fluorescent protein (GFP) from the jellyfish Aequorea. A core question in the biochemistry of DsRed is the mechanism by which the GFP-like 475-nm excitation and 500-nm emission maxima of immature DsRed are red-shifted to the 558-nm excitation and 583-nm emission maxima of mature DsRed. After digestion of mature DsRed with lysyl endopeptidase, high-resolution mass spectra of the purified chromophore-bearing peptide reveal that some of the molecules have lost 2 Da relative to the peptide analogously prepared from a mutant, K83R, that stays green. Tandem mass spectrometry indicates that the bond between the alpha-carbon and nitrogen of Gln-66 has been dehydrogenated in DsRed, extending the GFP chromophore by forming C==N==C==O at the 2-position of the imidazolidinone. This acylimine substituent quantitatively accounts for the red shift according to quantum mechanical calculations. Reversible hydration of the C==N bond in the acylimine would explain why denaturation shifts mature DsRed back to a GFP-like absorbance. The C==N bond hydrolyses upon boiling, explaining why DsRed shows two fragment bands on SDS/PAGE. This assay suggests that conversion from green to red chromophores remains incomplete even after prolonged aging. PMID- 11050231 TI - Molecular spectroscopy and dynamics of intrinsically fluorescent proteins: coral red (dsRed) and yellow (Citrine). AB - Gene expression of intrinsically fluorescent proteins in biological systems offers new noninvasive windows into cellular function, but optimization of these probes relies on understanding their molecular spectroscopy, dynamics, and structure. Here, the photophysics of red fluorescent protein (dsRed) from discosoma (coral), providing desired longer emission/absorption wavelengths, and an improved yellow fluorescent protein mutant (Citrine) (S65G/V68L/Q69 M/S72A/T203Y) for significant comparison, are characterized by using fluorescence correlation spectroscopy and time-correlated single-photon counting. dsRed fluorescence decays as a single exponential with a 3.65 +/- 0.07-ns time constant, indicating a single emitting state/species independent of pH 4.4-9.0, in contrast with Citrine. However, laser excitation drives reversible fluorescence flicker at 10(3)-10(4) Hz between dark and bright states with a constant partition fraction f(1) = 0.42 +/- 0.06 and quantum yield of approximately 3 x 10(-3). Unlike Citrine (pKa approximately 5.7), pH-dependent proton binding is negligible (pH 3. 9-11) in dsRed. Time-resolved anisotropy of dsRed reveals rapid depolarization (211 +/- 6 ps) plus slow rotational motion (53 +/- 8 ns), in contrast with a single rotational time (16 +/- 2 ns) for Citrine. The molecular dimensions, calculated from rotational and translational diffusion, indicate that dsRed is hydrodynamically 3.8 +/- 0.4 times larger than predicted for a monomer, which suggests an oligomer (possibly a tetramer) configuration even at approximately 10(-9) M. The fast depolarization is attributed to intraoligomer energy transfer between mobile nonparallel chromophores with the initial anisotropy implying a 24 +/- 3 degrees depolarization angle. Large two photon excitation cross sections ( approximately 100 GM at 990 nm for dsRed and approximately 50 GM at 970 nm for Citrine), advantageous for two-photon fluorescence imaging in cells, are measured. PMID- 11050232 TI - Replication by a single DNA polymerase of a stretched single-stranded DNA. AB - A new approach to the study of DNA/protein interactions has been opened through the recent advances in the manipulation of single DNA molecules. These allow the behavior of individual molecular motors to be studied under load and compared with bulk measurements. One example of such a motor is the DNA polymerase, which replicates DNA. We measured the replication rate by a single enzyme of a stretched single strand of DNA. The marked difference between the elasticity of single- and double-stranded DNA allows for the monitoring of replication in real time. We have found that the rate of replication depends strongly on the stretching force applied to the template. In particular, by varying the load we determined that the biochemical steps limiting replication are coupled to movement. The replication rate increases at low forces, decreases at forces greater than 4 pN, and ceases when the single-stranded DNA substrate is under a load greater than approximately 20 pN. The decay of the replication rate follows an Arrhenius law and indicates that multiple bases on the template strand are involved in the rate-limiting step of each cycle. This observation is consistent with the induced-fit mechanism for error detection during replication. PMID- 11050233 TI - Nonglassy kinetics in the folding of a simple single-domain protein. AB - Theory suggests that the otherwise rapid folding of simple heteropolymer models becomes "glassy"-dominated by multiple kinetically trapped misfolded states-at low temperatures or when the overall bias toward the native state is reduced relative to the depth of local minima. Experimental observations of nonsingle exponential protein-folding kinetics have been taken as evidence that the protein folding free energy landscape is similarly rough. No equivalent analysis, however, has been reported for a simple single-domain protein lacking prolines, disulfide bonds, prosthetic groups, or other gross structural features that might complicate folding. In an effort to characterize the glassiness of a folding free energy landscape in the absence of these potentially complicating factors, we have monitored the folding of a kinetically simple protein, peptostreptococcal protein L (protein L). We observe no statistically significant deviation from homogeneous single-exponential relaxation kinetics across temperatures ranging from near the protein's melting temperature to as low as -15 degrees C. On the basis of these observations, we estimate that, if there is a glass transition in the folding of protein L, it occurs at least 45 degrees C and possibly more than 145 degrees C below the freezing point of water. Apparently the folding free energy landscape of protein L is extremely smooth, which may be indicative of a rate-limiting step in folding that is, effectively, a nonglassy process. PMID- 11050234 TI - Anatomy of protein structures: visualizing how a one-dimensional protein chain folds into a three-dimensional shape. AB - Here, we depict the anatomy of protein structures in terms of the protein folding process. Via an iterative, top-down dissecting procedure, tertiary structures are spliced down to reveal their anatomy: first, to produce domains (defined by visual three-dimensional inspection criteria); then, hydrophobic folding units (HFU); and, at the end of a multilevel process, a set of building blocks. The resulting anatomy tree organization not only clearly depicts the organization of a one-dimensional polypeptide chain in three-dimensional space but also straightforwardly describes the most likely folding pathway(s). Comparison of the tree with the formation of the hydrophobic folding units through combinatorial assembly of the building blocks illustrates how the chain folds in a sequential or a complex folding pathway. Further, the tree points to the kinetics of the folding, whether the chain is a fast or a slow folder, and the probability of misfolding. Our ability to successfully dissect the protein into an anatomy tree illustrates that protein folding is a hierarchical process and further validates a building blocks protein folding model. PMID- 11050235 TI - Peg3/Pw1 promotes p53-mediated apoptosis by inducing Bax translocation from cytosol to mitochondria. AB - Mitochondria is believed to play a central role in p53-mediated apoptosis. However, the signal transduction pathways leading to mitochondria remain unclear. Here, we report that translocation of Bax protein from cytosol to mitochondria is required for p53-induced apoptosis. Cytosolic Bax is unable to induce apoptosis, and blocking Bax translocation inhibits cell death. Expression of Bcl-2 blocks cytochrome c release and apoptosis but has no effect on Bax translocation, suggesting that Bax translocation acts upstream of Bcl-2. We further demonstrate that Peg3/Pw1, a protein up-regulated in p53-mediated cell death process, induces Bax translocation independent of apoptosis. The results suggest that Bax translocation represents an important regulatory step in p53-mediated apoptosis, and Peg3/Pw1 functions as a modulator downstream of p53 to regulate Bax redistribution in the cells, thus favoring the cellular decision toward apoptosis over growth arrest following p53 induction. PMID- 11050236 TI - Hematopoietic reconstitution of SLP-76 corrects hemostasis and platelet signaling through alpha IIb beta 3 and collagen receptors. AB - Mice deficient in the hematopoietic cell-specific adapter protein SLP-76 demonstrate a failure of T cell development and fetal hemorrhage. Although SLP-76 deficient platelets manifest defective collagen receptor signaling, this alone may not explain the observed bleeding diathesis. Because alpha IIb beta 3, the platelet fibrinogen receptor, is required for normal hemostasis, we explored a potential role for SLP-76 in alpha IIb beta 3 signaling. Interaction of soluble or immobilized fibrinogen with normal human or murine platelets triggers rapid tyrosine phosphorylation of SLP-76. Moreover, platelet adhesion to fibrinogen stimulates actin rearrangements, filopodial and lamellipodial extension, and localization of tyrosine phosphorylated proteins to the cell periphery. In contrast, SLP-76-deficient murine platelets bind fibrinogen normally, but spread poorly and exhibit reduced levels of phosphotyrosine. The in vivo bleeding diathesis as well as the defects in platelet responses to fibrinogen and collagen are reversed by retroviral transduction of SLP-76 into bone marrow derived from SLP-76-deficient mice. These studies establish that SLP-76 functions downstream of alpha IIb beta 3 and collagen receptors in platelets. Furthermore, expression of SLP-76 in hematopoietic cells, including platelets, plays a necessary role in hemostasis. PMID- 11050238 TI - Skeletal muscle deformity and neuronal disorder in Trio exchange factor-deficient mouse embryos. AB - Dbl-homology guanine nucleotide exchange factors (DH-GEFs) regulate actin cytoskeletal reorganization, cell adhesion, and gene transcription via activation of Rho GTPases. However, little is known about the physiological role of mammalian DH-GEFs during development. The DH-GEF family member Trio is of particular interest because it is a multifunctional protein possessing two GEF domains, as well as a protein serine/threonine kinase domain, and trio-like genes in Caenorhabditis elegans and Drosophila were shown to function in neural migration and axon guidance. To determine the role of Trio during mammalian development, we generated a mouse trio loss-of-function mutation (trio(-/-)). Trio function is essential during late embryonic development as genotype analysis indicated that trio(-/-) embryos died between embryonic day (E)-15.5 and birth, or shortly thereafter. In the trio(-/-) embryos, primary skeletal myofibers were relatively normal at E14.5, but by E18.5 highly unusual spherical myofibers accumulated. Trio deficiency may cause a defect in secondary myogenesis, as the appearance of the abnormal trio(-/-) skeletal myofibers temporally coincided with the onset of secondary myogenesis, and smaller secondary myofibers located adjacent to the primary myofibers were absent. The proliferation of trio(-/-) secondary myoblasts appeared normal, suggesting that Trio may regulate secondary myoblast alignment or fusion. trio(-/-) embryos also displayed aberrant organization in several regions within the brain, including the hippocampal formation and olfactory bulb. We thus conclude that Trio is essential for late embryonic development, and that Trio functions in fetal skeletal muscle formation and in the organization of neural tissues. PMID- 11050237 TI - Creatine kinase, an ATP-generating enzyme, is required for thrombin receptor signaling to the cytoskeleton. AB - Thrombin orchestrates cellular events after injury to the vascular system and extravasation of blood into surrounding tissues. The pathophysiological response to thrombin is mediated by protease-activated receptor-1 (PAR-1), a seven transmembrane G protein-coupled receptor expressed in the nervous system that is identical to the thrombin receptor in platelets, fibroblasts, and endothelial cells. Once activated by thrombin, PAR-1 induces rapid and dramatic changes in cell morphology, notably the retraction of growth cones, axons, and dendrites in neurons and processes in astrocytes. The signal is conveyed by a series of localized ATP-dependent reactions directed to the actin cytoskeleton. How cells meet the dynamic and localized energy demands during signal transmission is unknown. Using the yeast two-hybrid system, we identified an interaction between PAR-1 cytoplasmic tail and the brain isoform of creatine kinase, a key ATP generating enzyme that regulates ATP within subcellular compartments. The interaction was confirmed in vitro and in vivo. Reducing creatine kinase levels or its ATP-generating potential inhibited PAR-1-mediated cellular shape changes as well as a PAR-1 signaling pathway involving the activation of RhoA, a small G protein that relays signals to the cytoskeleton. Thrombin-stimulated intracellular calcium release was not affected. Our results suggest that creatine kinase is bound to PAR-1 where it may be poised to provide bursts of site specific high-energy phosphate necessary for efficient receptor signal transduction during cytoskeletal reorganization. PMID- 11050239 TI - Direct genetic demonstration of G alpha 13 coupling to the orphan G protein coupled receptor G2A leading to RhoA-dependent actin rearrangement. AB - G2A is an orphan G protein-coupled receptor (GPCR), expressed predominantly in T and B cells and homologous to a small group of GPCRs of unknown function expressed in lymphoid tissues. G2A is transcriptionally induced in response to diverse stimuli, and its ectopic expression suppresses transformation of B lymphoid precursors by BCR-ABL. G2A induces morphological transformation of NIH 3T3 fibroblasts. Microinjection of constructs encoding G2A into Swiss 3T3 fibroblasts induces actin reorganization into stress fibers that depends on RhoA, but not CDC42 or RAC. G2A elicits RhoA-dependent transcriptional activation of serum response factor. Direct evaluation of RhoA activity demonstrates elevated levels of RhoA-GTP in G2A-expressing cells. Microinjection of embryonic fibroblasts derived from various G alpha knockout mice establishes a requirement for G alpha 13 but not G alpha 12 or G alpha q/11 in G2A-induced actin rearrangement. In conclusion, G2A represents a family of GPCRs expressed in lymphocytes that may link diverse stimuli to cytoskeletal reorganization and transcriptional activation through a pathway involving G alpha 13 and RhoA. PMID- 11050240 TI - Antagonistic role of vega1 and bozozok/dharma homeobox genes in organizer formation. AB - During zebrafish development, zygotic gene expression initiated at the midblastula transition converts maternal information on embryo polarity into a transcriptional read-out. Expression of a homeobox gene, vega1, is activated at midblastula transition in all blastomeres, but is down-regulated dorsally before gastrulation. Ubiquitous expression of vega1 is maintained in bozozok mutants, in which the dorsal-specific homeobox gene bozozok/dharma (boz/dha) is disrupted and organizer formation is impaired. Vega1 inhibits expression of boz/dha and organizer-specific genes, and causes ventralization resulting in a headless phenotype. In contrast, VP16-vega1, a fusion including the Vega1 homeodomain and VP16 activation domain, elicits ectopic expression of organizer genes and suppresses several aspects of the boz mutant phenotype. We propose that boz/dha dependent down-regulation of vega1 in the dorsal region is an early essential step in organizer formation in zebrafish. PMID- 11050241 TI - Parameters of self-organization in Hydra aggregates. AB - Self-organization has been demonstrated in a variety of systems ranging from chemical-molecular to ecosystem levels, and evidence is accumulating that it is also fundamental for animal development. Yet, self-organization can be approached experimentally in only a few animal systems. Cells isolated from the simple metazoan Hydra can aggregate and form a complete animal by self-organization. By using this experimental system, we found that clusters of 5-15 epithelial cells are necessary and sufficient to form de novo head-organizing centers in an aggregate. Such organizers presumably arise by a community effect from a small number of cells that express the conserved HyBra1 and HyWnt genes. These local sources then act to pattern and instruct the surrounding cells as well as generate a field of lateral inhibition that ranges up to 1,000 microm. We propose that conserved patterning systems in higher animals originate from extremely robust and flexible molecular self-organizing systems that were selected for during early metazoan evolution. PMID- 11050242 TI - Clonogenic hepatoblasts, common precursors for hepatocytic and biliary lineages, are lacking classical major histocompatibility complex class I antigen. AB - An in vitro colony-forming assay and flow cytometry were used to identify rat hepatoblasts as being classical MHC class I, RT1A(l-), OX18(low) intercellular adhesion molecule 1 (ICAM-1)(+). Inducible differentiation toward biliary lineage was observed in most colonies derived from single RT1A(l-) progenitors, proving their bipotentiality. These findings demonstrate the antigenic profile of clonogenic hepatoblasts and proof of their bipotency. Furthermore, whereas colony formation of adult hepatocytes required epidermal growth factor, clonal growth of hepatoblasts was potentiated without epidermal growth factor. The adult hepatic colonies consisted of RT1A(l+)OX18(+)ICAM-1(++) cells. These results indicate that hepatoblasts possess unique characteristics as compared with adult hepatocytes harboring significant proliferative activity. The phenotypic identity of hepatoblasts and the clonal culture system have relevance for identifying hepatic stem cells from adults, for studying liver development, and for cell therapy based on hepatic progenitors. PMID- 11050243 TI - PCGEM1, a prostate-specific gene, is overexpressed in prostate cancer. AB - A prostate-specific gene, PCGEM1, was identified by differential display analysis of paired normal and prostate cancer tissues. Multiple tissue Northern blot analysis revealed that PCGEM1 was expressed exclusively in human prostate tissue. Analysis of PCGEM1 expression in matched normal and primary tumor specimens revealed tumor-associated overexpression in 84% of patients with prostate cancer by in situ hybridization assay and in 56% of patients by reverse transcription PCR assay. Among various prostate cancer cell lines analyzed, PCGEM1 expression was detected only in the androgen receptor-positive cell line LNCaP. Extensive DNA sequence analysis of the PCGEM1 cDNA and genomic DNA revealed that PCGEM1 lacks protein-coding capacity and suggests that it may belong to an emerging class of noncoding RNAs, also called "riboregulators." The PCGEM1 locus was mapped to chromosome 2q32. Taken together, the remarkable prostate-tissue specificity and androgen-dependent expression of PCGEM1 as well as its elevated expression in a significant percentage of tumor tissues suggest specific functions of PCGEM1 in the biology and tumorigenesis of the prostate gland. PMID- 11050244 TI - Hyperglycemia-induced mitochondrial superoxide overproduction activates the hexosamine pathway and induces plasminogen activator inhibitor-1 expression by increasing Sp1 glycosylation. AB - The hexosamine pathway has been implicated in the pathogenesis of diabetic complications. We determined first that hyperglycemia induced a decrease in glyceraldehyde-3-phosphate dehydrogenase activity in bovine aortic endothelial cells via increased production of mitochondrial superoxide and a concomitant 2.4 fold increase in hexosamine pathway activity. Both decreased glyceraldehyde-3 phosphate dehydrogenase activity and increased hexosamine pathway activity were prevented completely by an inhibitor of electron transport complex II (thenoyltrifluoroacetone), an uncoupler of oxidative phosphorylation (carbonyl cyanide m-chlorophenylhydrazone), a superoxide dismutase mimetic [manganese (III) tetrakis(4-benzoic acid) porphyrin], overexpression of either uncoupling protein 1 or manganese superoxide dismutase, and azaserine, an inhibitor of the rate limiting enzyme in the hexosamine pathway (glutamine:fructose-6-phosphate amidotransferase). Immunoprecipitation of Sp1 followed by Western blotting with antibodies to O-linked GlcNAc, phosphoserine, and phosphothreonine showed that hyperglycemia increased GlcNAc by 1.7-fold, decreased phosphoserine by 80%, and decreased phosphothreonine by 70%. The same inhibitors prevented all these changes. Hyperglycemia increased expression from a transforming growth factor beta(1) promoter luciferase reporter by 2-fold and increased expression from a ( 740 to +44) plasminogen activator inhibitor-1 promoter luciferase reporter gene by nearly 3-fold. Inhibition of mitochondrial superoxide production or the glucosamine pathway prevented all these changes. Hyperglycemia increased expression from an 85-bp truncated plasminogen activator inhibitor-1 (PAI-1) promoter luciferase reporter containing two Sp1 sites in a similar fashion (3.8 fold). In contrast, hyperglycemia had no effect when the two Sp1 sites were mutated. Thus, hyperglycemia-induced mitochondrial superoxide overproduction increases hexosamine synthesis and O-glycosylation of Sp1, which activates expression of genes that contribute to the pathogenesis of diabetic complications. PMID- 11050245 TI - Hematopoietic progenitor cells grow on 3T3 fibroblast monolayers that overexpress growth arrest-specific gene-6 (GAS6). AB - Pluripotential hematopoietic stem cells grow in close association with bone marrow stromal cells, which play a critical role in sustaining hematopoiesis in long-term bone marrow cultures. The mechanisms through which stromal cells act to support pluripotential hematopoietic stem cells are largely unknown. This study demonstrates that growth arrest-specific gene-6 (GAS6) plays an important role in this process. GAS6 is a ligand for the Axl (Ufo/Ark), Sky (Dtk/Tyro3/Rse/Brt/Tif), and Mer (Eyk) family of tyrosine kinase receptors and binds to these receptors via tandem G domains at its C terminus. After translation, GAS6 moves to the lumen of the endoplasmic reticulum, where it is extensively gamma-carboxylated. The carboxylation process is vitamin K dependent, and current evidence suggests that GAS6 must be gamma-carboxylated to bind and activate any of the cognate tyrosine kinase receptors. Here, we show that expression of GAS6 is highly correlated with the capacity of bone marrow stromal cells to support hematopoiesis in culture. Nonsupportive stromal cell lines express little to no GAS6, whereas supportive cell lines express high levels of GAS6. Transfection of the cDNA encoding GAS6 into 3T3 fibroblasts is sufficient to render this previously nonsupportive cell line capable of supporting long-term hematopoietic cultures. 3T3 cells, genetically engineered to stably express GAS6 (GAS6-3T3), produce a stromal layer that supports the generation of colony forming units in culture (CFU-c) for up to 6 wk. Hematopoietic support by genetically engineered 3T3 is not vitamin K dependent, and soluble recombinant GAS6 does not substitute for coculturing the hematopoietic progenitors with genetically modified 3T3 cells. PMID- 11050246 TI - Intracellular antibody-caspase-mediated cell killing: an approach for application in cancer therapy. AB - Antibodies have been expressed inside cells in an attempt to ablate the function of oncogene products. To make intracellular antibodies more generally applicable and effective in cancer therapy, we have devised a method in which programmed cell death or apoptosis can be triggered by specific antibody-antigen interaction. When intracellular antibodies are linked to caspase 3, the "executioner" in the apoptosis pathway, and bind to the target antigen, the caspase 3 moieties are self-activated and thereby induce cell killing. We have used this strategy in a model system with two pairs of intracellular antibodies and antigens. In vivo coexpression of an antibody-caspase 3 fusion with its antigenic target induced apoptosis that was specific for antibody, antigen, and active caspase 3. Moreover, the antibody-caspase 3 fusion protein was not toxic to cells in the absence of antigen. Therefore, intracellular antibody-mediated apoptosis should be useful as a specific therapeutic approach for the treatment of cancers, a situation where target cell killing is required. PMID- 11050247 TI - Visualizing gene expression by whole-body fluorescence imaging. AB - Transgene expression in intact animals now can be visualized by noninvasive techniques. However, the instruments and protocols developed so far have been formidable and expensive. We describe here a system for rapidly visualizing transgene expression in major organs of intact live mice that is simple, rapid, and eminently affordable. Green fluorescent protein (GFP) is expressed in the cells of brain, liver, pancreas, prostate, and bone, and its fluorescence is encoded in whole-body optical images. For low-magnification images, animals are illuminated atop a fluorescence light box and directly viewed with a thermoelectrically cooled color charge-coupled device camera. Higher magnification images are made with the camera focused through an epi-fluorescence dissecting microscope. Both nude and normal mice were labeled by directly injecting 8 x 10(10) plaque-forming units/ml of adenoviral GFP in 20-100 microl PBS and 10% glycerol into either the brain, liver, pancreas, prostate, or bone marrow. Within 5-8 h after adenoviral GFP injection, the fluorescence of the expressed GFP in brain and liver became visible, and whole-body images were recorded at video rates. The GFP fluorescence continued to increase for at least 12 h and remained detectable in liver for up to 4 months. The system's rapidity of image acquisition makes it capable of real-time recording. It requires neither exogenous contrast agents, radioactive substrates, nor long processing times. The method requires only that the expressed gene or promoter be fused or operatively linked to GFP. A comparatively modest investment allows the study of the therapeutic and diagnostic potential of suitably tagged genes in relatively opaque organisms. PMID- 11050248 TI - A secreted Salmonella protein induces a proinflammatory response in epithelial cells, which promotes neutrophil migration. AB - In response to Salmonella typhimurium, the intestinal epithelium generates an intense inflammatory response consisting largely of polymorphonuclear leukocytes (neutrophils, PMN) migrating toward and ultimately across the epithelial monolayer into the intestinal lumen. It has been shown that bacterial-epithelial cell interactions elicit the production of inflammatory regulators that promote transepithelial PMN migration. Although S. typhimurium can enter intestinal epithelial cells, bacterial internalization is not required for the signaling mechanisms that induce PMN movement. Here, we sought to determine which S. typhimurium factors and intestinal epithelial signaling pathways elicit the production of PMN chemoattractants by enterocytes. Our results suggest that S. typhimurium activates a protein kinase C-dependent signal transduction pathway that orchestrates transepithelial PMN movement. We show that the type III effector protein, SipA, is not only necessary but is sufficient to induce this proinflammatory response in epithelial cells. Our results force us to reconsider the long-held view that Salmonella effector proteins must be directly delivered into host cells from bacterial cells. PMID- 11050249 TI - Reduction of axonal caliber does not alleviate motor neuron disease caused by mutant superoxide dismutase 1. AB - It is well established that motor neurons with large axon caliber are selectively affected in amyotrophic lateral sclerosis (ALS). To investigate whether high neurofilament (NF) content and large axonal caliber are factors that predispose motor neurons to selective degeneration in ALS, we generated mice expressing a mutant form of superoxide dismutase 1 (SOD1(G37R)) linked to familial ALS in a context of one allele for each NF gene being disrupted. A approximately 40% decrease of NF protein content detected in triple heterozygous knockout mice shifted the calibers of large axons in L5 ventral root from 5-9 microm to 1-5 microm, altering neither the normal subunit stoichiometry and morphological distribution of NFs nor levels of other cytoskeletal proteins. This considerable reduction in NF burden and caliber of axons did not extend the life span of SOD1(G37R) mice nor did it alleviate the loss of motor axons. Moreover, increasing the density of NFs in axons by overexpressing a NF-L transgene did not accelerate disease in SOD1(G37R) mice. These results do not support the current view that high NF content and large caliber of axons may account for the selective vulnerability of motor neurons in ALS caused by mutant SOD1. PMID- 11050250 TI - Development and persistence of kindling epilepsy are impaired in mice lacking glial cell line-derived neurotrophic factor family receptor alpha 2. AB - Seizure activity regulates gene expression for glial cell line-derived neurotrophic factor (GDNF) and neurturin (NRTN), and their receptor components, the transmembrane c-Ret tyrosine kinase and the glycosylphosphatidylinositol anchored GDNF family receptor (GFR) alpha 1 and alpha 2 in limbic structures. We demonstrate here that epileptogenesis, as assessed in the hippocampal kindling model, is markedly suppressed in mice lacking GFR alpha 2. Moreover, at 6 to 8 wk after having reached the epileptic state, the hyperexcitability is lower in GFR alpha 2 knock-out mice as compared with wild-type mice. These results provide evidence that signaling through GFR alpha 2 is involved in mechanisms regulating the development and persistence of kindling epilepsy. Our data suggest that GDNF and NRTN may modulate seizure susceptibility by altering the function of hilar neuropeptide Y-containing interneurons and entorhinal cortical afferents at dentate granule cell synapses. PMID- 11050251 TI - Constitutive expression of the neuron-restrictive silencer factor (NRSF)/REST in differentiating neurons disrupts neuronal gene expression and causes axon pathfinding errors in vivo. AB - The neuron-restrictive silencer factor (NRSF; also known as REST for repressor element-1 silencing transcription factor) is a transcriptional repressor of multiple neuronal genes, but little is known about its function in vivo. NRSF is normally down-regulated upon neuronal differentiation. Constitutive expression of NRSF in the developing spinal cord of chicken embryos caused repression of two endogenous target genes, N-tubulin and Ng-CAM, but did not prevent overt neurogenesis. Nevertheless, commissural neurons that differentiated while constitutively expressing NRSF showed a significantly increased frequency of axon guidance errors. These data suggest that down-regulation of NRSF is necessary for the proper development of at least some classes of neurons in vivo. PMID- 11050252 TI - Role of microtubules in the intracellular distribution of tobacco mosaic virus movement protein. AB - Despite its central role in virus infection, little is known about the mechanisms of intracellular trafficking of virus components within infected cells. In this study, we followed the dynamics of tobacco mosaic virus movement protein (MP) distribution in living protoplasts after disruption of microtubules (MTs) by cold treatment and subsequent rewarming to 29 degrees C. At early stages of infection, cold treatment (4 degrees C) caused the accumulation of MP fused to green fluorescent protein (GFP) in large virus replication bodies that localized in perinuclear positions, whereas at midstages of infection, the association of MP:GFP with MTs was disrupted. Rewarming the protoplasts to 29 degrees C reestablished the association of MTs with the replication bodies that subsequently spread throughout the cytoplasm and to the periphery of the cell. The role of MTs in the intracellular distribution of the MP also was analyzed by examining the distribution pattern of a nonfunctional mutant of MP (TAD5). Like MP:GFP, TAD5:GFP interacted with the endoplasmic reticulum membranes and colocalized with its viral RNA but did not colocalize with MTs. The involvement of MTs in the intracellular distribution of tobacco mosaic virus MP is discussed. PMID- 11050253 TI - Failure to handle more than one internal representation in visual detection tasks. AB - Perceptual studies make a clear distinction between sensitivity and decision criterion. The former is taken to characterize the processing efficiency of the underlying sensory system and it increases with stimulus strength. The latter is regarded as the manifestation of a subjective operation whereby individuals decide on (as opposed to react reflexively to) the occurrence of an event based on factors such as expectation and payoff, in addition to its strength. To do so, individuals need to have some knowledge of the internal response distributions evoked by this event or its absence. In a natural, behaviorally relevant multistimulus environment, observers must handle many such independent distributions to optimize their decision criteria. Here we show that they cannot do so. Instead, while leaving sensitivity unchanged, lower and higher visibility events tend to be reported respectively less and more frequently than when they are presented in isolation. This behavior is in quantitative agreement with predictions based on the notion that observers represent a multistimulus environment as a unitary internal distribution to which each stimulus contributes proportionally to its probability of occurrence. Perceptual phenomena such as blindsight, hemineglect, and extinction may be, at least in part, accounted for in such a way. PMID- 11050254 TI - Assisted discharge for patients with exacerbations of chronic obstructive pulmonary disease: safe and effective. PMID- 11050255 TI - Inhaled disodium cromoglycate as maintenance therapy for childhood asthma: time to consign to history? PMID- 11050256 TI - Control and prevention of tuberculosis in the United Kingdom: code of practice 2000. Joint Tuberculosis Committee of the British Thoracic Society. AB - BACKGROUND: The guidelines on control and prevention of tuberculosis in the United Kingdom have been reviewed and updated. METHODS: A subcommittee was appointed by the Joint Tuberculosis Committee (JTC) of the British Thoracic Society to revise the guidelines published in 1994 by the JTC, including representatives of the Royal College of Nursing, Public Health Medicine Environmental Group, and Medical Society for Study of Venereal Diseases. In preparing the revised guidelines the authors took account of new published evidence and graded the strength of evidence for their recommendations. The guidelines have been approved by the JTC and the Standards of Care Committee of the British Thoracic Society. RECOMMENDATIONS: Tuberculosis services in each district should have staffing and resources to fulfil both the control and prevention recommendations in this document and to ensure adequate treatment monitoring. Notification of tuberculosis is required for surveillance and to initiate contact tracing (where appropriate). The following areas are discussed and recommendations made where appropriate: (1) public health law in relation to tuberculosis; (2) the organisational requirements for tuberculosis services; (3) measures for control of tuberculosis in hospitals, including segregation of patients; (4) the requirements for health care worker protection, including HIV infected health care workers; (5) measures for control of tuberculosis in prisons; (6) protection for other groups with potential exposure to tuberculosis; (7) awareness of the high rates of tuberculosis in the homeless together with local plans for detection and action; (8) detailed advice on contact tracing; (9) contact tracing required for close contacts of bovine tuberculosis; (10) management of tuberculosis in schools; (11) screening of new immigrants and how this should be performed; (12) outbreak contingency investigation; and (13) BCG vaccination and the management of positive reactors found in the schools programme. PMID- 11050257 TI - Early discharge for patients with exacerbations of chronic obstructive pulmonary disease: a randomized controlled trial. AB - BACKGROUND: We have previously reported the use of a hospital based respiratory nurse service (Acute Respiratory Assessment Service, ARAS) to support home treatment of patients with exacerbations of chronic obstructive pulmonary disease (COPD). A controlled trial was undertaken to compare early discharge with home treatment supported by respiratory nurses with conventional hospital management of patients admitted with exacerbations of COPD. METHODS: Patients with COPD admitted as emergencies were identified the next working day. They were eligible for inclusion in the study if the differential diagnosis included an exacerbation of COPD, but were excluded if other medical conditions or acidotic respiratory failure required inpatient investigation or management. Of 360 patients reviewed, 209 were being assessed for other active medical problems and were excluded, 33 potential participants were already involved in research studies and so were ineligible, and 37 did not wish to participate in the study. Eighty one patients were randomised to receive conventional inpatient care (n=40) or to planned early discharge the next working day (n=41). Those discharged early continued treatment at home under the supervision of specialist respiratory nurses. Outcome measures were readmission, additional hospital days, and deaths within 60 days of initial admission. Process measures included number of visits, duration of follow up by the respiratory nurse, and additional treatment provided to support early discharge. RESULTS: On an intention to treat basis, a policy of early discharge reduced inpatient stay from a mean of 6.1 (range 1-13) days with conventional management to 3.2 (1-16) days with an early discharge policy. Twelve patients (30% conventional management, 29.3% early discharge) were readmitted in each group giving a mean difference in readmission of 0.7% (95% CI of the difference 19.2 to 20.6). In the conventional management group readmitted patients spent a mean of 8.75 additional days in hospital compared with 7.83 days in the early discharge group, giving a mean difference of 0.92 days (95% CI of the difference 6.5 to 8.3). There were two deaths (5%) in the conventional management group and one (2.4%) in the early discharge group, a mean difference of 2.6% (95% CI of the difference -5.7 to 10.8). CONCLUSIONS: Patients with acute exacerbations of COPD uncomplicated by acidotic respiratory failure or other medical problems can be discharged home earlier than is current practice with support by visiting respiratory nurses. No difference was found in the subsequent need for readmission. PMID- 11050258 TI - Randomized controlled trial of supported discharge in patients with exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: A randomised trial was performed on patients presenting to hospital with an exacerbation of chronic obstructive pulmonary disease (COPD) to compare outcomes in those managed at home with support with those admitted to hospital in the standard manner. METHODS: Over an 18 month period all patients presenting to the Royal Infirmary of Edinburgh on weekdays (n=718) with a diagnosis of an exacerbation of COPD were assessed for inclusion in the trial. Patients with impaired level of consciousness, acute confusion, acute changes on radiography, or an arterial pH of <7.35 or with other serious medical or social reasons for admission were excluded. Patients randomised to home support were discharged with an appropriate treatment package (antibiotics, corticosteroids, nebulised bronchodilators and, if necessary, home oxygen). They were visited by a nurse the following day and thereafter at intervals of 2-3 days until recovery when they were discharged from follow up. Parallel observations were made on patients allocated to normal hospital admission up to the point of discharge. Patients in both groups were assessed at home eight weeks after the initial assessment. RESULTS: Among weekday patients 353 (50%) were considered obligatory admissions, 140 (19%) were admitted because of co-morbidity, 17 (2%) because of poor social circumstances, and 24 (3%) did not consent to the trial. The remaining 184 (26%) were randomised (2:1) either to home support or to a standard hospital admission. The median time to discharge was 7 days for the home support group and 5 days for the admitted group (p<0.01); 25% of the home support group and 34% of the admitted group were readmitted before the final assessment at eight weeks (p>0.05). There were no significant differences between the groups in attendances by GPs and carers or in health status measured eight weeks after the initial assessment. Satisfaction with the service was good. The mean total health service cost per patient was estimated as 877 pounds sterling for the home support group and 1753 pounds sterling for the admitted group. CONCLUSIONS: This study shows that home supported discharge is a well tolerated, safe, and economic alternative to hospital admission for a proportion of patients referred to hospital for admission for an exacerbation of COPD. PMID- 11050259 TI - Inhaled disodium cromoglycate (DSCG) as maintenance therapy in children with asthma: a systematic review. AB - BACKGROUND: Disodium cromoglycate (DSCG) is included in the BTS guidelines on the treatment of asthma for use in children, but is now used only infrequently. We have identified and interpreted the findings of all published randomised, placebo controlled trials of DSCG in the prophylactic treatment of children with asthma. METHODS: Several databases were searched to identify trials. Studies were included if they investigated subjects with asthma aged 0-18 years old, addressed maintenance treatment with inhaled DSCG, and were published in English. The methodological quality of the studies was assessed independently by three reviewers. The 95% confidence intervals (CI) of differences in the treatment effect for cough and wheeze between placebo and treatment with DSCG were computed. The estimates were pooled and tested for homogeneity and, to assess possible publication bias, a funnel plot was made and tested for symmetry. RESULTS: Of the 24 randomised, placebo controlled trials identified, the methodological scores varied widely. The null hypothesis of homogeneity was rejected. Under the assumption of heterogeneity the overall CI for wheeze was 0.11 to 0.26 and for cough was 0.13 to 0.27. The overall tolerance intervals ( 0.11 to 0. 48 and -0.04 to 0.43 for wheeze and cough, respectively) both included zero, so it cannot be concluded that future studies will show an effect of DSCG compared with placebo. Older studies were more often in favour of DSCG. The funnel plots suggest publication bias; small studies with negative or equal outcomes are lacking. CONCLUSION: Given the apparent publication bias, the small overall treatment effect, and the tolerance interval including zero, there is insufficient evidence that DSCG has a beneficial effect as maintenance treatment in children with asthma. PMID- 11050260 TI - Intranasal challenge with aspirin in the diagnosis of aspirin intolerant asthma: evaluation of nasal response by acoustic rhinometry. AB - BACKGROUND: Nasal provocation tests with lysine-aspirin have recently been introduced for assessment of aspirin intolerant asthma. A study was undertaken to evaluate the usefulness of acoustic rhinometry, a new non-invasive technique, in the diagnosis of aspirin intolerant asthma/rhinitis. METHODS: Fifteen patients with aspirin intolerant asthma/rhinitis (nine women, mean (SD) age 54.7 (14) years), eight patients with aspirin tolerant asthma/rhinitis (three women, mean (SD) age 52.6 (7.8) years), and eight healthy subjects (two women, mean (SD) age 32.5 (9.7) years) were studied. All subjects were challenged with saline (0.9% NaCl) and 25 mg lysine acetylsalicylic acid (L-ASA) instilled into each nostril of the nose on two separate days. The clinical response was evaluated based on nasal symptoms (sneezes, itching, secretion and blockage). The nasal response was measured by acoustic rhinometry. Symptoms and rhinometry curves were recorded at 10 minute intervals for three hours, one hour before challenge and two hours after challenge. RESULTS: L-ASA challenge induced a significant increase in symptoms in patients with aspirin intolerant asthma/rhinitis. No differences in the clinical response were detected in those with aspirin tolerant asthma/rhinitis or healthy subjects. L-ASA challenge induced a significant decrease in nasal volume measured by acoustic rhinometry in aspirin intolerant patients. No differences were detected between the challenges in aspirin tolerant patients. If a 25% decrease in nasal volume is taken as the cut off point, the specificity of the test was 94% and the sensitivity reached 73%. The nasal challenge was well tolerated by all subjects. CONCLUSION: Acoustic rhinometry may be used to study the nasal response to L-ASA. Nasal challenge with L-ASA is safe and can be used as a diagnostic test even in asthmatic patients with severe bronchial obstruction. PMID- 11050261 TI - Herbal medicines for asthma: a systematic review. AB - BACKGROUND: Asthma is one of the most common chronic diseases in modern society and there is increasing evidence to suggest that its incidence and severity are increasing. There is a high prevalence of usage of complementary medicine for asthma. Herbal preparations have been cited as the third most popular complementary treatment modality by British asthma sufferers. This study was undertaken to determine if there is any evidence for the clinical efficacy of herbal preparations for the treatment of asthma symptoms. METHODS: Four independent literature searches were performed on Medline, Pubmed, Cochrane Library, and Embase. Only randomised clinical trials were included. There were no restrictions on the language of publication. The data were extracted in a standardised, predefined manner and assessed critically. RESULTS: Seventeen randomised clinical trials were found, six of which concerned the use of traditional Chinese herbal medicine and eight described traditional Indian medicine, of which five investigated Tylophora indica. Three other randomised trials tested a Japanese Kampo medicine, marihuana, and dried ivy leaf extract. Nine of the 17 trials reported a clinically relevant improvement in lung function and/or symptom scores. CONCLUSIONS: No definitive evidence for any of the herbal preparations emerged. Considering the popularity of herbal medicine with asthma patients, there is urgent need for stringently designed clinically relevant randomised clinical trials for herbal preparations in the treatment of asthma. PMID- 11050262 TI - Personal exposure to nitrogen dioxide and risk of airflow obstruction in asthmatic children with upper respiratory infection. AB - BACKGROUND: Several studies have linked air pollution by nitrogen dioxide (NO(2)) with increased hospital admissions for asthma in children. Exacerbations of asthma in children are often precipitated by upper respiratory infections. It is therefore possible that NO(2) increases the risk of airways obstruction when asthmatic children develop upper respiratory infections. METHODS: To test this hypothesis a sample of 114 asthmatic children aged 7-12 years were followed for a total of up to 13 months. Probable upper respiratory infections were identified by consensus review of daily symptom diaries, and episodes of airways obstruction from serial records of peak expiratory flow (PEF). Personal exposures to NO(2) were measured with Palmes tubes that were changed weekly. Generalised estimating equations were used to assess the relative risk (RR) of an asthmatic exacerbation starting within seven days of an upper respiratory infection according to estimated NO(2) exposure during the one week period from two days before to four days after the onset of the infection. RESULTS: The children were followed for an average of 34 weeks during which 318 upper respiratory infections and 224 episodes of reduced PEF were diagnosed. PEF episodes were much more likely to occur in the seven days following the onset of an upper respiratory infection than at other times. Estimated exposures to NO(2) at the time of infections were generally low (geometric mean 10.6 microg/m(3)). Compared with exposures of < or = 8 microg/m(3), exposures of >28 microg/m(3) were associated with a RR of 1.9 (95% confidence interval 1.1 to 3.4) for the development of an asthmatic episode within seven days of an infection. CONCLUSIONS: The findings give some support to the hypothesis that NO(2) increases the risk of asthmatic exacerbations following respiratory infections, even at relatively low levels of exposure. Further studies in populations with higher exposures would be useful. PMID- 11050263 TI - Pulmonary arterial hypertension in patients with sleep apnoea syndrome. AB - BACKGROUND: Pulmonary arterial hypertension (PAH) in patients with sleep apnoea syndrome (SAS) is classically ascribed to associated chronic obstructive pulmonary disease (COPD). The aim of this retrospective study was to evaluate the possible occurrence of PAH as a complication of SAS in patients without COPD. METHODS: Right heart catheterisation was performed in 44 patients with SAS and without COPD confirmed by polysomnography (apnoea index >5/h) admitted for the administration of nasal continuous positive airway pressure (CPAP). RESULTS: Precapillary PAH, defined as mean pulmonary arterial pressure of >20 mm Hg with pulmonary capillary wedge pressure <15 mm Hg, was observed in 12/44 (27%) patients with SAS. There were no significant differences in apnoea index between patients with (PAH+) and those without PAH (PAH-) (42.6 (26.3) versus 35.8 (21.7) apnoeas/h). The PAH+ group differed significantly from the PAH- group in the following respects: lower daytime arterial oxygen tension (PaO(2)) (9.6 (1.1) versus 11.3 (1.5) kPa, p=0.0006); higher daytime arterial carbon dioxide tension (PaCO(2)) (5.8 (0.5) versus 5.3 (0.5) kPa, p=0.002); more severe nocturnal hypoxaemia with a higher percentage of total sleep time spent at SaO(2) <80% (32.2 (28.5)% versus 10.7 (18.8)%, p=0.005); and higher body mass index (BMI) (37.4 (6) versus 30.3 (6.7) kg/m(2), p=0.002). The PAH+ patients had significantly lower values of vital capacity (VC) (87 (14)% predicted versus 105 (20)% predicted, p=0.005), forced expiratory volume in one second (FEV(1)) (82 (14)% predicted versus 101 (17)% predicted, p=0.001), expiratory reserve volume (40 (16)% predicted versus 77 (41)% predicted, p=0.003), and total lung capacity (87 (13)% predicted versus 98 (18)% predicted, p=0.04). Stepwise multiple regression analysis showed that mean pulmonary artery pressure (PAPm) was positively correlated with BMI and negatively with PaO(2). CONCLUSION: Pulmonary arterial hypertension is frequently observed in patients with SAS, even when COPD is absent, and appears to be related to the severity of obesity and its respiratory mechanical consequences. PMID- 11050265 TI - Development of a disease specific health related quality of life measure for adults and adolescents with cystic fibrosis. AB - BACKGROUND: Health related quality of life (HRQoL) measurement is important in determining the impact of disease on daily functioning and subsequently informing interventions. In cystic fibrosis (CF) generic HRQoL measures have been employed but these may not be sufficiently specific. The aim of the current work was to develop and validate a disease specific HRQoL measure for adults and adolescents with cystic fibrosis. METHODS: Areas of concern to adults and adolescents with CF were identified by unstructured interviews, self-administered questionnaires, consultation with multidisciplinary specialist staff, a review of the relevant literature, and examination of other HRQoL measures. Items for the questionnaire were generated on the basis of this process. Continued evaluation and development of the Cystic Fibrosis Quality of Life (CFQoL) questionnaire was undertaken by a process of statistical analysis and continued feedback from patients. The full testing and validation of the CFQoL questionnaire took place over four phases: (1) initial item generation and testing of a preliminary questionnaire, (2) testing and validation of the second version of the questionnaire, (3) test retest reliability of a third and final version of the questionnaire, and (4) sensitivity testing of the final version of the questionnaire. RESULTS: Nine domains of functioning were identified using principal components analysis with varimax rotation. Internal reliability of the identified domains was demonstrated using Cronbach alpha coefficients (range 0.72-0.92) and item to total domain score correlations. Concurrent validity (range r = 0.64-0.74), discriminatory ability between different levels of disease severity, sensitivity across transient changes in health (effect size range, moderate d = 0.56 to large d = 1.95), and test-retest reliability (r = 0.74-0.96) were also found to be robust. CONCLUSIONS: The CFQoL questionnaire is a fully validated disease specific measure consisting of 52 items across nine domains of functioning which have been identified by, and are of importance to, adolescents and adults with cystic fibrosis. This measure will be useful in clinical trials and longitudinal studies. PMID- 11050264 TI - Ventilatory responses to hypercapnia and hypoxia in relatives of patients with the obesity hypoventilation syndrome. AB - BACKGROUND: It is unclear why some morbidly obese individuals have waking alveolar hypoventilation while others with similar obesity do not. Some evidence suggests that patients with the obesity hypoventilation syndrome (OHS) may have a measurable premorbid impairment of ventilatory chemoresponsiveness. Such an impairment of ventilatory chemoresponsiveness in OHS, however, may be an acquired and reversible consequence of severe obstructive sleep apnoea (OSA). We hypothesised that, in patients with OHS who do not have coincident severe OSA, there may be a familial impairment in ventilatory responses to hypoxia and hypercapnia. METHODS: Sixteen first degree relatives of seven patients with OHS without severe OSA (mean (SD) age 40 (16) years, body mass index (BMI) 30 (6) kg/m(2)) and 16 subjects matched for age and BMI without OHS or OSA were studied. Selection criteria included normal arterial blood gas tensions and lung function tests and absence of sleep apnoea on overnight polysomnography. Ventilatory responses to isocapnic hypoxia and to hyperoxic hypercapnia were compared between the two groups. RESULTS: The slope of the ventilatory response to hypercapnia was similar in the relatives (mean 2.33 l/min/mm Hg) and in the control subjects (2.12 l/min/mm Hg), mean difference 0.2 l/min/mm Hg, 95% confidence interval (CI) for the difference -0.5 to 0.9 l/min/mm Hg, p=0.5. The hypoxic ventilatory response was also similar between the two groups (slope factor A: 379.1 l/min * mm Hg for relatives and 373.4 l/min * mm Hg for controls; mean difference 5.7 l/min * mm Hg; 95% CI -282 to 293 l/min * mm Hg, p=0.7; slope of the linear regression line of the fall in oxygen saturation and increase in minute ventilation: 2.01 l/min/% desaturation in relatives, 1.15 l/min/% desaturation in controls; mean difference 0. 5 l/min/% desaturation; 95% CI -1.7 to 0.7 l/min/% desaturation, p=0. 8). CONCLUSION: There is no evidence of impaired ventilatory chemoresponsiveness in first degree relatives of patients with OHS compared with age and BMI matched control subjects. PMID- 11050266 TI - The ligase chain reaction as a primary screening tool for the detection of culture positive tuberculosis. AB - BACKGROUND: The ligase chain reaction Mycobacterium tuberculosis assay uses ligase chain reaction technology to detect tuberculous DNA sequences in clinical specimens. A study was undertaken to determine its sensitivity and specificity as a primary screening tool for the detection of culture positive tuberculosis. METHODS: The study was conducted on 2420 clinical specimens (sputum, bronchoalveolar lavage fluid, pleural fluid, urine) submitted for primary screening for Mycobacterium tuberculosis to a regional medical microbiology laboratory. Specimens were tested in parallel with smear, ligase chain reaction, and culture. RESULTS: Thirty nine patients had specimens testing positive by the ligase chain reaction assay. Thirty two patients had newly diagnosed tuberculosis, one had a tuberculosis relapse, three had tuberculosis (on antituberculous therapy when tested), and three had healed tuberculosis. In the newly diagnosed group specimens were smear positive in 21 cases (66%), ligase chain reaction positive in 30 cases (94%), and culture positive in 32 cases (100%). Using a positive culture to diagnose active tuberculosis, the ligase chain reaction assay had a sensitivity of 93.9%, a specificity of 99.8%, a positive predictive value of 83.8%, and a negative predictive value of 99.9%. CONCLUSIONS: This study is the largest clinical trial to date to report the efficacy of the ligase chain reaction as a primary screening tool to detect Mycobacterium tuberculosis infection. The authors conclude that ligase chain reaction is a useful primary screening test for tuberculosis, offering speed and discrimination in the early stages of diagnosis and complementing traditional smear and culture techniques. PMID- 11050267 TI - Involvement of Epstein-Barr virus latent membrane protein 1 in disease progression in patients with idiopathic pulmonary fibrosis. AB - BACKGROUND: The role of Epstein-Barr virus (EBV) in idiopathic pulmonary fibrosis (IPF) is uncertain. A study was undertaken to detect the virus in IPF as well as to clarify the influence of EBV on the clinical features of the disease. METHODS: Twenty nine lung specimens were obtained from patients with IPF, as well as five specimens from patients with systemic sclerosis with pulmonary fibrosis (SSc) and 15 specimens from controls. EBV DNA and EBV latent membrane protein 1 (LMP1) were detected using the PCR method and immunohistochemical analysis, respectively. RESULTS: EBV DNA was detected in 24 of 25 patients with IPF (96%), in all five patients with SSc (100%), and in 10 of 14 controls (71%). The detection ratio was significantly higher in patients with IPF than in controls (p = 0.047, odds ratio (OR) = 9.60, 95% confidence interval (CI) 0.9 to 96.9). Immunohistochemical analysis revealed that cuboidal epithelial cells were positively stained with anti-LMP1 antibody in nine of the 29 lung specimens from IPF patients. In contrast, neither the patients with SSc nor the control subjects showed positive staining. In the follow up periods LMP1 positive patients with IPF died more frequently from respiratory failure than LMP1 negative patients (4/9 versus 1/20; p = 0.022, OR = 15.20, 95% CI 1. 3 to 168.0). CONCLUSIONS: EBV LMP1 positivity may be associated with more rapid disease progression in IPF. PMID- 11050268 TI - Resource implications of patients with multidrug resistant tuberculosis. AB - BACKGROUND: Multidrug resistant tuberculosis (MDR TB) requires a complex drug regimen and lengthy multidisciplinary care. The financial cost of successful management of each case is potentially large. METHODS: The costs of managing nine HIV negative patients with pulmonary MDR TB were compared with 18 age group and ethnicity matched controls with fully sensitive disease. Calculations included: cost of outpatient visits and inpatient stays including negative pressure isolation; costs of drug provision and toxicity monitoring; costs of additional procedures and multidisciplinary referrals. RESULTS: The mean cost of managing a case of pulmonary MDR TB was in excess of 60,000 pounds sterling and for sensitive disease it was 6040 pounds sterling. CONCLUSIONS: Clinicians and healthcare commissioning authorities may both be underestimating the costs of managing MDR TB, and accordingly the consequences for units dealing with such cases may be serious. Funding of care for MDR TB in the UK requires strategic decisions at regional or governmental level. PMID- 11050269 TI - Pediatric origins of adult lung diseases. 3: the genesis of adult sleep apnoea in childhood. PMID- 11050271 TI - [Adenotomy and adenotonsillectomy--useful in recurrent acute otitis media?]. PMID- 11050272 TI - Change in Managing Editor. PMID- 11050270 TI - Factors influencing airway inflammation in chronic obstructive pulmonary disease. PMID- 11050274 TI - Pharmacologic management of arrhythmias. AB - The role of antiarrhythmic drugs in the management of children with arrhythmias has changed due to the rapid development of radiofrequency ablation. Moreover, the release of new drugs and a better insight into the electrophysiologic mechanisms of arrhythmias have changed former patterns of drug management. However, because of lack of controlled trials, arrhythmia management in pediatrics is still mainly based on clinical studies and individual experience. Within these limitations, I attempt to give practical recommendations for the management of the different arrhythmias seen in children based on the latest developments in this area. PMID- 11050275 TI - Usefulness of the electrocardiogram in diagnosing mechanisms of tachycardia. AB - The electrocardiogram, despite its simplistic technological composition, remains a valuable tool in the diagnosis of pediatric arrythmias. In this article the characteristic features of different tachycardias are reviewed. PMID- 11050276 TI - Current approach to pediatric syncope. AB - Syncope is the abrupt loss of consciousness and postural tone that reverses without intervention. Typically, syncopal episodes in pediatric patients are brief, lasting seconds (rarely minutes), and are followed by complete recovery without residual neurological sequelae. Syncopal presentations may be dramatic and lead family members and primary care providers to suspect a malignant cardiac condition, prompting referral to a pediatric cardiologist. Significant advances in the understanding of syncope in infants, children, and adolescents have occurred in the past decade. This review emphasizes neurally mediated syncope, but other etiologies are reviewed to complete the spectrum encountered by pediatric cardiologists. Some clues are provided to distinguish the more common and benign forms of syncope from those due to significant underlying heart disease. PMID- 11050277 TI - Ventricular arrhythmias: when to worry. AB - Although isolated premature ventricular contractions may be seen in as many as 15% of normal newborns, one third of normal adolescents, and two thirds of adolescents and adults with repaired heart disease, sustained ventricular arrhythmias are relatively rare in young normal hearts. Sudden cardiac health is rare in young normal hearts, although there is an increased incidence in dilated cardiomyopathies and following repair of particular congenital heart lesions. Noninvasive and invasive techniques imperfectly stratify these patients. Patients with cardiomyopathy often have ventricular arrhythmias, although the risk of mortality is more closely linked to ventricular function. There are many infants and pediatric patients with apparently normal hearts who have combinations of asymptomatic nonsustained ventricular tachycardia and potentially serious symptoms. The clinical concern is to identify diagnoses such as long QT syndrome associated with recurrent cardiac syncope and premature mortality so that appropriate choices can be made regarding drug and device therapy. Although this broad range of disease places a premium on careful evaluation, selective therapy, and continued research, serious symptoms, even in the absence of ectopy, are concerning in any patient. PMID- 11050278 TI - Current concepts in long QT syndrome. AB - Sudden cardiac death occurs in the United States with an incidence of more than 300,000 persons per year. The underlying cause of death is commonly considered to be due to primary or secondary arrhythmias. In young persons in whom no structural heart disease can be identified, the long QT syndromes (LQTS) are commonly considered as likely causes. Multiple genes causing LQTS have been identified thus far, all of which encode cardiac ion channels. These include two potassium channel alpha subunits (KVLQT1 and HERG), two potassium channel beta subunits (minK and MiRP1), and one sodium channel gene (SCN5A). The purpose of this review is to describe the current understanding of the molecular genetics of LQTS and the resultant phenotypes, particularly in young patients. PMID- 11050279 TI - Radiofrequency catheter ablation: indications and complications. AB - Radiofrequency catheter ablation was first described in pediatric patients in the early 1990s. Since then, multiple advances in the technology and understanding of radiofrequency ablation have allowed this technique to blossom into one of the most powerful therapeutic tools available to the pediatric electrophysiologist. This treatment has, in the majority of cases, replaced arrhythmia surgery as the definitive cure for most arrhythmias. Ablation therapy is commonly implemented as an elective procedure to treat paroxysmal reentrant supraventricular tachycardia. There are several advantages to this therapy when used in the common indications: no exercise restrictions, no need for chronic drug therapy, and the avoidance of hospital visits for breakthrough episodes. This review will discuss the indications for radiofrequency ablation in the current era. In order to fully discuss this issue, this review will include the prior treatment of arrhythmias, current success rates, complications, and potential long-term issues. PMID- 11050280 TI - Radiofrequency catheter ablation of tachycardia in patients with congenital heart disease. AB - Patients with anomalies of the heart frequently suffer from arrhythmias that either are associated with a congenital heart defect or result from the course of the disease. For most of the bradyarrhythmias, appropriate timing of the initiation of treatment is more challenging than its eventual execution. In the case of tachycardias, technical aspects of treatment require more attention because the often imperative impact such tachycardias have on quality of life, morbidity, and mortality determine intervention timing. Increasingly, interventional electrophysiology is turned to as a potentially definitive and substrate-related treatment because of antiarrhythmic drug therapy's failure to prevent arrhythmia recurrences and the potential detrimental side effects from drug therapy seen in this particular patient population. Using the experience gained during the past 10 years in the treatment of patients with arrhythmias but without associated structural heart disease, several groups reported their results and difficulties with the application of such therapy to patients with congenital heart defects. In this report, we summarize our hospital's experience with transcatheter radiofrequency current application for treatment of various types of tachyarrhythmias in 139 children and adults with congenital heart defects, emphasizing the current limitations of such therapy and addressing the potential benefits expected from future technology. Patient ages ranged from 5 months to 76 years (mean 25.3 +/- 17.7 years), including 56 children and adolescents less than 16 years of age. At least one attempt at surgical palliation or correction was made in 93 patients; the remaining 46 patients had no surgical intervention attempts. A total of 225 different tachycardias were found, 93 of which were based on a congenital arrhythmogenic substrate (e.g., an accessory pathway). Acquired substrates (e.g., scars or myocardial fibrosis) gave rise to the remaining 132 tachycardias. Radiofrequency current ablation (183 sessions) successfully treated 121 of 139 patients. Within a follow-up period of 21 months a recurrence of the intrinsically treated tachycardia was seen in 24 patients (10.7%); 13 of the 24 underwent a successful repeat session. There were no significant procedure-related complications. Young and adult patients with congenital heart disease can be safely and successfully treated for tachycardias with the use of radiofrequency current ablation. Because such treatment meets the specific needs of this patient group, early consideration for this therapy is recommended. PMID- 11050281 TI - New directions in surgical therapy of arrhythmias. AB - The success of the radiofrequency catheter ablation procedure for most types of supraventricular and ventricular tachycardia, particularly in young patients, largely eliminated the role of surgical therapy of arrhythmias. However, there remains a subset of arrhythmia patients in whom the catheter approach has not been successful and types of arrhythmias with high recurrence rates following initially successful catheter ablation procedures where surgery can provide more definitive therapy. In addition, the concepts of ablation therapy can be successfully incorporated into the concomitant repair of complex congenital heart disease, resulting in single-stage therapy for structural and rhythm abnormalities. Prospectively, knowledge of the role of anatomic barriers as substrates for future reentrant arrhythmia circuits provides the opportunity to alter these circuits prophylactically at the time of initial surgical repair of congenital heart disease in an attempt to avoid the late development of tachycardia. This article describes our experience during the past decade with 71 patients undergoing arrhythmia surgery using this approach. PMID- 11050283 TI - From other journals PMID- 11050285 TI - The endothelin system in septic and endotoxin shock. AB - The view of the endothelium as a passive barrier has gradually changed as a number of endothelium-derived substances have been discovered. Substances like nitric oxide, prostaglandins and endothelins have potent and important properties, involving not only the circulation as such but also the response to stimuli like inflammation and trauma. The endothelin system, discovered in 1988, has not only strong vasoconstrictor properties, but also immunomodulating, endocrinological and neurological effects exerted through at least two types of receptors. Septic shock, a condition with high mortality, is associated with vast cardiovascular changes, organ dysfunction with microcirculatory disturbances and dysoxia. In the experimental setting, endotoxaemia resembles these changes and is, as well as septic shock, accompanied by a pronounced increase in plasma endothelin levels. The pathophysiology in septic and endotoxin shock remains to be fully elucidated, but several studies indicate that endothelial dysfunction is one contributing mechanism. Activation of the endothelin system is associated with several pathological conditions complicating septic shock, such as acute respiratory distress syndrome, cardiac dysfunction, splanchnic hypoperfusion and disseminated intravascular coagulation. Through the development of both selective and nonselective endothelin receptor antagonists, the endothelin system has been the object of a large number of studies during the last decade. This review highlights systematically the findings of previous studies in the area. It provides strong indications that the endothelin system, apart from being a marker of vascular injury, is directly involved in the pathophysiology of septic and endotoxin shock. Interventions with endothelin receptor antagonists during septic and endotoxin shock have so far only been done in animal studies but the results are interesting and promising. PMID- 11050286 TI - Mitogen-activated protein kinase inhibitors suppress prostaglandin F(2alpha) induced myosin-light chain phosphorylation and contraction in iris sphincter smooth muscle. AB - The purpose of this study was to investigate the potential role of mitogen activated protein (MAP) kinase in contraction by monitoring MAP kinase phosphorylation (activation) and contraction during agonist stimulation of cat iris sphincter smooth muscle. Changes in tension in response to prostaglandin F(2alpha), latanoprost, a prostaglandin F(2alpha) analog used as an anti-glaucoma drug, and carbachol were recorded isometrically, and MAP kinase activation was monitored by Western blot using a phosphospecific p42/p44 MAP kinase antibody. We found that treatment of the muscle with 2'-Amino-3'-methoxyflavone (PD98059) (10 microM), a specific inhibitor of MAP kinase kinase (MEK), inhibited significantly prostaglandin F(2alpha)- and latanoprost-induced phosphorylation and contraction, but had little effect on those evoked by carbachol. Prostaglandin F(2alpha) increased MAP kinase phosphorylation in a concentration-dependent manner with EC(50) value of 1.1 x 10(-8) M and increased contraction with EC(50) of 0.92 x 10(-9) M. The MAP kinase inhibitors PD98059, Apigenin and 1,4-Diamino-2,3-dicyano 1, 4bis(2-aminophenylthio)butadiene (UO126) inhibited prostaglandin F(2alpha) induced contraction in a concentration-dependent manner with IC(50) values of 2.4, 3.0 and 4.8 microM, respectively. PD98059 had no effect on prostaglandin F(2alpha)- or on carbachol-stimulated inositol-1,4,5-trisphosphate (IP(3)) production. In contrast, the MAP kinase inhibitor inhibited prostaglandin F(2alpha)-induced myosin-light chain (MLC) phosphorylation, but had no effect on that of carbachol. N-[2-(N-(4-Chloro-cinnamyl)-N-methylaminomethyl)phenyl]-N-[2- hydroxyethyl]-4-methoxybenzenesulfonamide (KN-93) (10 microM), a Ca(2+) calmodulin-dependent protein kinase inhibitor, and Wortmannin (10 microM), an MLC kinase inhibitor, inhibited significantly (by 80%) prostaglandin F(2alpha)- and carbachol-induced contraction. It can be concluded that in this smooth muscle p42/p44 MAP kinases are involved in the mechanism of prostaglandin F(2alpha)-, but not in that of carbachol, induced contraction. In addition, these data clearly indicate that the stimulation of the iris sphincter with prostaglandin F(2alpha) and carbachol activate two distinct pathways, the MAP kinase pathway and the Ca(2+) mobilization pathway. PMID- 11050287 TI - On the significance of telomerase activity in human malignant glioma cells. AB - Telomerase is critical for tumor cell immortalization and is a novel target for cancer chemotherapy. Here, we examined whether telomerase is expressed in glioma cell lines, whether telomerase activity is regulated by bcl-2 or p53, and whether telomerase activity predicts response to chemotherapy. Further, we characterized the effects of a candidate telomerase inhibitor, penclomedine, in glioma cells. All 12 human malignant glioma cell lines examined were telomerase positive. Telomerase activity was not modulated during cell cycle progression, did not correlate with p53 status or bcl-2 family protein expression, and did not predict drug sensitivity, except for an association with resistance to carmustine. Ectopic bcl-2 expression did not enhance telomerase activity. Wild-type p53 reduced telomerase activity in cell lines retaining p53 activity but not in p53 mutant cell lines. Penclomedine killed glioma cells via an apoptotic, but death receptor-, bcl-2- and caspase-independent pathway, but did not inhibit telomerase and did not act synergistically with cytotoxic drugs. We conclude that telomerase activity does not account for the differential chemosensitivity of human glioma cells and that penclomedine kills glioma cells via a telomerase-independent pathway. PMID- 11050288 TI - LY367265, an inhibitor of the 5-hydroxytryptamine transporter and 5 hydroxytryptamine(2A) receptor antagonist: a comparison with the antidepressant, nefazodone. AB - The potential antidepressant, LY367265 (1-[2-[4-(6-fluoro-1H-indol-3-yl)-3, 6 dihydro-1(2H)-pyridinyl]ethyl]-5,6-dihydro-1H,4H-[1,2, 5]thiadiazolo[4.3.2 ij]quinoline-2,2,-dioxide) has been shown to have a higher affinity for the 5 hydroxytryptamine (5-HT) transporter (K(i)=2.3 nM) and 5-HT(2A) (K(i)=0.81 nM) receptor than the clinically effective antidepressant, nefazodone. It is a potent inhibitor of [3H]5-HT uptake into rat cortical synaptosomes (IC(50)=3.1 nM) and shows selectivity over that for [3H]noradrenaline (IC(50)>1000 nM). It potentiates potassium-induced [3H]5-HT outflow from prelabelled guinea pig cortical slices both in the presence (EC(50)=950 nM) and absence (EC(50)=250 nM) of a saturating concentration of the 5-HT transport inhibitor, paroxetine, indicating a low level of activity at the 5-HT(1B/1D) autoreceptor. These studies indicate that LY367265 is a putative antidepressant which, because of its 5 HT(2A) receptor antagonist activity, has the potential to produce less sleep disturbance and sexual dysfunction than selective serotonin uptake inhibitors. PMID- 11050289 TI - Evidence for antagonist activity of the dopamine D3 receptor partial agonist, BP 897, at human dopamine D3 receptor. AB - The dopaminergic system has long been implicated in the mechanisms of reward and addiction. 1-(4-(2-Naphthoylamino)butyl)-4-(2-methoxyphenyl)-1A-piperazine HCl (BP 897) has been claimed to be a selective dopamine D3 receptor partial agonist and has recently been shown to inhibit cocaine-seeking behaviour, suggesting a role for dopamine D3 receptor agonists in the treatment of addiction. We have previously characterised the pharmacological profile of the human dopamine D3 and D2(long) receptors using microphysiometry and radioligand binding and we have now studied the interaction of BP 897 with the dopamine D2 and D3 receptors using these methods. At both human dopamine D3 and D2 receptors, BP 897 lacked agonist activity but was a potent and selective antagonist with pK(b) values of 8.05+/ 0.16 (4) and 9.43+/-0.22 (4) at human dopamine D2 and D3 receptors, respectively. These results, therefore, suggest that it may be the dopamine D3 receptor antagonist properties of BP 897 which have potential in the treatment of addiction and withdrawal. PMID- 11050290 TI - Potent inhibition of a recombinant low voltage-activated Ca(2+) channel by SB 209712. AB - T-type Ca(2+) currents were recorded in 2 mM Ca(2+) from HEK293 cells stably expressing the low voltage-activated Ca(2+) channel sub-unit alpha(1I). These currents were inhibited by the known Ca(2+) channel antagonist mibefradil with an IC(50) close to 1 microM. SB-209712 (1,6,bis?1-[4-(3 phenylpropyl)piperidinyl]?hexane), a compound originally developed as a high voltage-activated Ca(2+) channel blocker, proved to be a more potent T-type channel antagonist, exhibiting an IC(50) in the region of 500 nM. The antagonism produced by SB-209712 was reversed following drug removal and the observed antagonism exhibited little or no voltage-dependence with respect to either holding or test potential. These data indicate that SB-209712 is amongst the most potent known non-peptide T-type channel antagonists and thus may have some use in understanding the role of these channels in cellular function. PMID- 11050291 TI - Ambroxol as an inhibitor of nitric oxide-dependent activation of soluble guanylate cyclase. AB - The influence of ambroxol on the activity of human platelet soluble guanylate cyclase and rat lung soluble guanylate cyclase was investigated. Ambroxol in the concentration range from 0.1 to 10 microM had no effect on the basal activity of both enzymes and slightly enhanced it at 50 and 100 microM. Ambroxol inhibited in a concentration-dependent manner the sodium nitroprusside-induced activation of both enzymes. The IC(50) values for inhibition by ambroxol of sodium nitroprusside-stimulated human platelet soluble guanylate cyclase and rat lung soluble guanylate cyclase were 3.9 and 2.1 microM, respectively. Ambroxol did not influence the stimulation of soluble guanylate cyclase by protoporphyrin 1X. Thus, it is possible that the molecular mechanism of the therapeutic action of ambroxol involves the inhibition of nitric oxide (NO)-dependent activation of soluble guanylate cyclase. PMID- 11050292 TI - GABA(B) receptor mechanism and imipramine-induced antinociception in ligated and non-ligated mice. AB - This study concerned the effects of GABA(B) receptor agents on imipramine-induced antinociception in ligated and non-ligated mice in hot-plate test. The data showed that different doses of morphine (3, 6 and 9 mg/kg) induced a dose dependent antinociception in non-ligated or ligated mice. However, the opioid response was decreased in the ligated animals. Intracerebroventricular (i.c.v.) administration of imipramine (5, 10, 20 and 40 microg/mouse) did not induce antinociception in either non-ligated or ligated mice. However, the response induced in the ligated mice was less than that induced in the non-ligated animals. Intraperitoneal (i.p.) administration of imipramine (10, 20, 30 and 40 mg/kg) induced antinociception in both ligated and non-ligated animals. The responses to the drug were not significantly different in the two groups. Administration of baclofen either i.c.v. (0.125, 0.25 and 0. 5 microg/mouse) or i.p. (0.5, 1, 2 and 4 mg/kg) induced antinociception. The response to the drug was not significantly different in ligated and non-ligated mice. I.c.v. administration of a lower dose of baclofen (0.125 microg/mouse) with different doses of imipramine (2.5, 5 and 10 mg/kg) potentiates the response of imipramine. This effect was reduced by i.c.v. injection of GABA(B) receptor antagonist, CGP35348 [P-(3-aminopropyl)-p-diethoxymethyl-phosphinic acid] (20 microg/mouse). The higher dose of antagonist (20 microg/mouse) also decreased the response induced by baclofen or imipramine. CGP35348 itself (2.5, 5, 10 and 20 microg/mouse) induced dose-dependent antinociception with no significant difference in the ligated and non-ligated mice. It is concluded that a GABA receptor mechanism(s) may modulate the antidepressant-induced antinociception. PMID- 11050293 TI - Different outcomes after acute and chronic treatment with nicotine in pre-pulse inhibition in Lister hooded rats. AB - Excessive tobacco consumption by schizophrenic patients may be a form of self medication, and nicotine in tobacco may alleviate deficits in information processing. We tested this hypothesis by determining whether nicotine (acute/chronic) would improve information processing in the rat using pre-pulse inhibition as a model. In study 1, rats were injected with nicotine 10 min prior to placement in startle chambers (0.001-0.1 or 0.03-0.3 mg/kg, s.c.). In study 2, rats were injected with either saline or nicotine (0.4 mg/kg, s.c.) for 21 consecutive days and assessed for locomotor activity, pre-pulse inhibition and changes in [3H]nicotine binding in whole brain. Acutely, nicotine had no effect on pre-pulse inhibition. By contrast, after chronic nicotine treatment, rats demonstrated a robust deficit in pre-pulse inhibition and significant increases in locomotor activity and [3H]nicotine binding. The deficit in pre-pulse inhibition after chronic treatment with nicotine may be the result of non specific behavioural activation due to increased mesolimbic dopamine release or, possibly, nicotine may rapidly desensitize nicotinic receptors important for normal information processing. PMID- 11050294 TI - Reciprocal regulation of nitric oxide and glutamate in the nucleus tractus solitarii of rats. AB - Nitric oxide (NO) and glutamate are both important mediators of the central cardiovascular regulation in the nucleus tractus solitarii. Our previous studies revealed that the central cardiovascular effects of NO in the nucleus tractus solitarii could be inhibited by glutamate receptor blockade. On the other hand, nitric oxide synthase (NOS) inhibitor attenuated the cardiovascular effects of glutamate. Thus, NO and glutamatergic systems appear to interact in central cardiovascular regulation. The present study examined whether NO and glutamate may affect each other's release/production in the nucleus tractus solitarii. A microdialysis probe was implanted into the nucleus tractus solitarii of male Sprague-Dawley rats, and the changes in the extracellular levels of glutamate and NO were determined by high performance liquid chromatography coupled with electrochemical detection and an NO analyzer, respectively. The results showed that NO solution elicited >10 fold increases in the extracellular level of glutamate, which returned to normal 60 min after the end of NO perfusion. The NO donor N-acetyl-penicillamine (SNAP) had an effect similar to NO solution. Furthermore, the glutamate level was reduced to 61% of basal value by perfusion with the NOS inhibitor, N(G)-monomethyl-L-arginine (L-NMMA). When glutamate receptor agonist N-methyl-D-aspartic acid (NMDA) or alpha-amino-3-hydroxy-5 methylixoxazole-4-propionic acid (AMPA) was administered into the nucleus tractus solitarii, the extracellular NO level was increased by 70-100%, whereas glutamate receptor antagonists (MK-801 hydrogen maleate and 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX)) did not alter the basal levels of NO. These results suggest that NO and glutamate may enhance each other's release/production in the nucleus tractus solitarii. This reciprocal regulation of NO and glutamate may be important in central cardiovascular control in the nucleus tractus solitarii. PMID- 11050295 TI - Effects of intravenous cocaine administration on cerebellar Purkinje cell activity. AB - The goal of the present study was to investigate the effects of intravenous cocaine administration on cerebellar Purkinje cell firing. Extracellular neuron activity was recorded and cells were locally excited with spaced microiontophoretic pulses of glutamate. Glutamate-evoked and spontaneous discharges were compared before and immediately following cocaine administration. Cocaine injections (1. 0 and 0.25 mg/kg, i.v.) induced a reversible suppression of both spontaneous activity and glutamate-evoked excitation. Procaine was ineffective in producing similar actions. Cocaine only inhibited glutamate induced excitation in animals pre-treated with reserpine (5 mg/kg, i.p.). Propranolol injections (10 mg/kg, i.p.) were ineffective in blocking cocaine induced inhibitions. Yohimbine (5 mg/kg, i.p.) pre-treatment abolished cocaine induced suppressions of either spontaneous or glutamate-evoked excitation. Therefore, cocaine administration decreases Purkinje cell spontaneous and glutamate-evoked discharges by a mechanism involving alpha(2)-adrenoceptor activation. It is suggested that by changing the normal function of the cerebellum cocaine can produce drug-related alterations in overt behavior and cognition. PMID- 11050296 TI - Effect of a selective 5-hydroxytryptamine reuptake inhibitor on brain extracellular noradrenaline: microdialysis studies using paroxetine. AB - The clinical efficacy of selective serotonin (5-hydroxytryptamine, 5-HT) reuptake inhibitors (SSRIs) is normally attributed to their ability to increase brain 5-HT function although recent preclinical findings indicate that their selectivity for 5-HT over noradrenaline may be less evident in vivo. The present study investigated the effects of the SSRI, paroxetine, on extracellular levels of noradrenaline. Microdialysis was carried out in the hippocampus of the awake rat. In rats treated twice daily for 14 days with paroxetine (5 mg/kg s.c.), dialysate levels of noradrenaline showed a maintained two-fold increase compared to saline injected controls. Paroxetine (5 mg/kg s.c.) administered once daily for 14 days did not cause a sustained increase in noradrenaline but levels showed a moderate (+58%) increase in response to a paroxetine challenge. Acute injection of paroxetine (5 mg/kg s.c.) did not elevate noradrenaline levels. Paroxetine (5 mg/kg s.c.) elevated dialysate 5-HT after both acute and repeated (twice daily for 14 days) treatment. The paroxetine-induced increase in noradrenaline (and 5 HT) was positively correlated with plasma concentrations of the drug, which were around the therapeutic range. In comparison to paroxetine, desipramine (10 mg/kg s.c.) caused a four-fold increase in dialysate noradrenaline (but did not change 5-HT) following repeated (once daily for 14 days) treatment and a two-fold increase at for acute treatment. In summary, despite its selectivity as a 5-HT reuptake inhibitor, paroxetine increased extracellular levels of noradrenaline in rat hippocampus following repeated administration. We discuss the possibility that a facilitation of noradrenaline function might be involved in the antidepressant effect of paroxetine, and possibly other SSRIs. PMID- 11050297 TI - Antinociceptive activity of ricinoleic acid, a capsaicin-like compound devoid of pungent properties. AB - The antinociceptive effect of ricinoleic acid ([R-(Z)]-12-hydroxy-9-octadecenoic acid) in comparison with capsaicin (trans-8-methyl-N-vanillyl-6-nonenamide) has been investigated in several "in vivo" tests. Acute topical application of capsaicin, but not ricinoleic acid, produced by itself an hyperalgesic effect detected as a decrease in paw withdrawal latency in response to a painful (heat) stimulus in mice. Capsaicin, but not ricinoleic acid at any dose tested, showed an irritant effect in the wiping test in guinea pig conjunctiva after local application and in the paw licking test in mice after intradermal injection. Whereas acute application of ricinoleic acid or capsaicin decreased paw withdrawal latency to heat in the presence of a pre-existing inflammation (injection of carrageenan in the mouse paw), the repeated local treatment for 8 days with either compounds markedly increased paw withdrawal latency. In a chronic model of inflammation (complete Freund's adjuvant arthritis in mice), the repeated topical and intradermal treatments with both ricinoleic acid and capsaicin increased paw withdrawal latency to heat, the antinociceptive effect of ricinoleic acid being more persistent than that of capsaicin. Antinociceptive effect of 8 days of treatment with ricinoleic acid and capsaicin was observed in acetic acid-induced writhing in mice, capsaicin-induced foot licking in mice and capsaicin-induced wiping movements in guinea pig conjunctiva. A decrease of substance P tissue levels in the mouse paw was found after repeated treatment with ricinoleic acid. In conclusion, ricinoleic acid seems to be a new antinociceptive agent lacking the pungent and acute hyperalgesic properties of capsaicin. PMID- 11050298 TI - In vivo histamine release from brain cortex: the effects of modulating cellular and extracellular sodium and calcium channels. AB - The in vivo mechanisms underlying the actions of modulating Na(+)- and Ca(2+) sensitive channels and its effect on basal histamine release in the cerebral cortex of freely-moving unanesthetized rats was investigated. Basal histamine release in the cerebral cortex was determined by in vivo microdialysis coupled with high-performance liquid chromatography (HPLC) fluorometry detection. Basal levels of histamine were 0.67+/-0.02 pmol/10 microl of dialysate. Diltiazem, a Ca(2+) channel antagonist, produced a dose-dependent decrease in dialysate basal histamine concentration. Elevated K(+) (100 mM) in the perfusion medium increased basal histamine to a maximum of 223% of the baseline value. Similarly, diltiazem (60 mM) reduced the K(+), veratridine (100 microg/ml) and ouabain (100 microM) evoked increase in dialysate histamine. Basal histamine decreased by 48% when the perfusate contained 3 microM of voltage dependent Na(+) antagonist tetrodotoxin. The results of these studies indicate that the release of histamine in rat cerebral cortex can be induced by modulating Na(+) and Ca(2+) channels and that the L-type voltage-dependent sensitive Ca(2+) channels are involved in this release process. PMID- 11050299 TI - Comparative actions of cibenzoline and disopyramide on I(Kr) and I(Ks) currents in rat sino-atrial nodal cells. AB - Modulation by class Ia antiarrhythmic drugs, cibenzoline and disopyramide, of the pacemaking activity and the underlying ionic currents in rat sino-atrial nodal cells was investigated using current-clamp and whole-cell patch-clamp techniques. Both drugs depressed the spontaneous activity and often caused sinus arrest. The negative chronotropic effect was significant at 10 microM cibenzoline and 30 microM disopyramide. The L-type Ca(2+) current (I(Ca)) and the hyperpolarization activated inward current decreased by 69.7+/-3.2% and by 45.8+/-3.0% at 30 microM cibenzoline and by 51. 2+/-3.3% and by 48.3+/-2.7% at 100 microM disopyramide, respectively. The delayed rectifier K(+) current, which is composed of rapidly and slowly activated currents (I(Kr) and I(Ks)), also decreased. The IC(50) values of I(Kr) for cibenzoline and disopyramide were 8.8+/-1. 1 and 25.1+/-2.3 microM, respectively. In the presence of 5 microM E-4031 (1-[2-(6-methyl-2 pyridyl)ethyl]-4-(4-methylsulfonylaminobenzoyl) piperidine), the IC(50) values of I(Ks) for cibenzoline and disopyramide were 12.3+/-1.8 and 81.1+/-2.3 microM, respectively. The I(Ks) was completely blocked by 30 microM 293B (trans-6-cyano-4 (N-ethylsulphonyl-N-methtamino)-3-hydroxy-2 , 2-dimethyl-chromane). These results indicate that the ionic currents are more sensitive to cibenzoline than disopyramide in rat sino-atrial nodal cells, and that I(Ca) and I(Kr) make major contributions to pacemaking activity. PMID- 11050300 TI - Activation of the retrohippocampal region in the rat causes dopamine release in the nucleus accumbens: disruption by fornix section. AB - There is a well-described projection from the retrohippocampus (subiculum and entorhinal cortex) to the nucleus accumbens that is involved in the control of psychomotor behaviour, and is implicated in the aetiology of schizophrenia. Cortical abnormalities are widely reported in the brains of schizophrenic patients, but it is unclear whether they are the cause or consequence of those changes in subcortical systems that are treated with neuroleptic drugs. We have, therefore, conducted a series of microdialysis experiments in anaesthetized rats to determine whether infusion of the excitotoxin, N-methyl-D-aspartate, into the retrohippocampus increases mesolimbic dopamine release. We found a clear and reproducible increase in extracellular dopamine in the nucleus accumbens following N-methyl-D-aspartate (2.5 microg), that was abolished when we sectioned the fimbria-fornix. Furthermore, inhibition of gamma-aminobutyric acid receptors following retrohippocampus administration of bicuculline (4 microg), also increased dopamine in the nucleus accumbens. The dopamine response to bicuculline was accompanied by an increase in dopamine metabolism, was long lasting, and also reduced by fornix section.The response to both N-methyl-D-aspartate and bicuculline depends on the integrity of the projection from the retrohippocampus to the nucleus accumbens. The results provide an underlying mechanism whereby a primary insult in the temporal cortex, caused by excessive N-methyl-D-aspartate receptor stimulation, can produce a hyperdopaminergic state. In addition, an increase in subiculo-accumbens activity evoked by bicuculline may also explain why patients with limbic epilepsy can develop a psychosis. PMID- 11050301 TI - Flumazenil prevents diazepam-elicited anticonvulsant action and concomitant attenuation of glutamate overflow. AB - Systemic administration of diazepam (5 mg/kg, i.p.) produced a prompt anticonvulsant effect in pilocarpine-induced seizures in freely moving rats. The anticonvulsant effect was associated with significant attenuation of pilocarpine evoked increases in extracellular hippocampal glutamate levels to below the baseline levels. The purpose of the present microdialysis study, therefore, was to investigate if the effect of diazepam on glutamate release was mediated at the level of the benzodiazepine gamma-aminobutyric acid(A) (GABA(A)) receptor complex to preclude any non-GABAergic mechanisms. Systemic administration of the specific benzodiazepine-receptor antagonist flumazenil (10 mg/kg, i.p. )-elicited complete reversal of diazepam-evoked anticonvulsant action and concomitant attenuation of extracellular glutamate efflux below the baseline levels. This provides evidence that under the given experimental conditions, diazepam-evoked alterations in glutamate overflow associated with the anticonvulsant action were indeed mediated at the level of benzodiazepine-GABA(A) receptor complex, possibly involving the modulation of both pre- and post-synaptic sites of the receptor complex. PMID- 11050302 TI - Involvement of adrenergic and cholinergic systems in nicotine-induced anxiogenesis in mice. AB - In the present study, the effect of alpha-adrenoceptor agents on response to nicotine in an anxiety model (elevated plus-maze) in mice was investigated. Administered nicotine reduced indices of anti-anxiety behaviour (percent open-arm time (%open-arm time) and percent open-arm entries (%open-arm entry)) and increased indices of anxiety behaviour (protected stretched attention posture and percent of protected head dipping (%protected dipping)), indicating that nicotine elicits an anxiogenic response. This response to the drug was obtained 7 min but not 30 min after drug injection and with doses of 0.25 and 0.5 mg/kg. Nicotinic receptor antagonists mecamylamine (0.5 and 1 mg/kg) and hexamethonium (5 and 10 mg/kg) reduced the response induced by nicotine (0.25 mg/kg). Mecamylamine (1 mg/kg; decreased %open-arm entry and increased protected stretched attention posture) and hexamethonium (10 mg/kg; decreased %open-arm time) showed an anxiogenic-like profile. A muscarinic receptor antagonist, atropine (2.5 and 5 mg/kg), did not alter the nicotine response but elicited an anxiogenic effect by itself. The alpha(1)-adrenoceptor antagonist prazosin (0.25 and 0.5 mg/kg), but not the alpha(1)-adrenoceptor agonist, phenylephrine (4 and 6 mg/kg), reversed the nicotine effect. Single administration of phenylephrine (6 mg/kg) increased %open-arm time, while prazosin did not alter the anxiety behaviour. The alpha(2) adrenoceptor agonist clonidine (0.001 and 0.01 mg/kg), induced complete immobility when administered in combination with nicotine. However, an alpha(2) adrenoceptor antagonist, yohimbine (0.5 and 1 mg/kg), appeared to reverse the nicotine response, but did not show interaction with nicotine's effect. Clonidine did not elicit any effect, but yohimbine (1 mg/kg) increased %open-arm entry and %open-arm time by itself. It can be concluded that certain doses of nicotine (0.25 and 0.5 mg/kg) 7 min after their injection induce an anxiogenic effect through nicotinic mechanism(s), and that involvement of alpha(1)- but not alpha(2)-adrenoceptors in the response to nicotine seems likely. PMID- 11050303 TI - The effects of some K(+) channel blockers on scopolamine- or electroconvulsive shock-induced amnesia in mice. AB - The effects of three K(+) channel blockers, 4-aminopyridine, 3, 4-diaminopyridine and apamin, on scopolamine- or electroconvulsive shock-induced amnesia were investigated in mice by using a one-trial step-down passive avoidance system. Scopolamine and electroconvulsive shock reduced the retention latency of passive avoidance, which indicated the amnestic effect of these treatments. 4 Aminopyridine, 3,4-diaminopyridine and apamin injected immediately after the acquisition trial, reversed the amnestic effect of scopolamine or electroconvulsive shock in a dose-dependent manner. None of the drugs or electroconvulsive shock treatment affected the rotarod or activity cage performance of the mice. These results indicate that K(+) channel blockers may improve cognitive deficits when memory is impaired by a drug or any other manipulation. PMID- 11050304 TI - Pharmacological analysis of contractile effects of eletriptan and sumatriptan on human isolated blood vessels. AB - Eletriptan, a second-generation triptan with high affinity for 5-HT(1B/1D) receptors, is highly effective in migraine, with or without aura. We compared the effects of eletriptan and sumatriptan on the human isolated middle meningeal and coronary arteries and saphenous vein, used as models for therapeutic efficacy and potential side effects, and have investigated the role of 5-HT(1B/1D) receptors in contractions induced by these triptans. Concentration-response curves to eletriptan and sumatriptan were constructed in the absence or presence of a selective 5-HT(1B/1D) receptor antagonist, N-[4-methoxy-3-(4-methylpiperazin-1 yl)phenyl]-3-methyl-4-(4-py rid yl) benzamide (GR125743). All three blood vessels constricted in response to eletriptan and sumatriptan, but the middle meningeal artery relaxed following the highest concentration (100 microM) of eletriptan. In the middle meningeal artery, GR125743 antagonised the contractions induced by both eletriptan (pEC(50): 7.34+/-0.13) and sumatriptan (pEC(50): 6.91+/-0.17) to a similar degree (pA(2): 8. 81+/-0.17 and 8.64+/-0.21, respectively). In the human coronary artery and saphenous vein, sumatriptan-induced contractions (pEC(50): 6.24+/-0.14 and 6.19+/-0.12, respectively) were also potently antagonised by GR125743 (pA(2): 8.18+/-0.27 and 8.34+/-0.12, respectively). The eletriptan-induced contractions of the human saphenous vein (pEC(50): 6.09+/ 0.13) were antagonised less effectively by GR125743 (pK(B): 7.73+/-0.18), and those of the human coronary artery (pEC(50): 5.54+/-0.22) remained unaffected by GR125743 up to a concentration of 100 nM. These results suggest that (i) based on the differences in pEC(50) values, the cranioselectivity of eletriptan (63-fold) is higher than that of sumatriptan (5-fold) in coronary artery, (ii) the contractile effects of sumatriptan and eletriptan (lower concentrations) in the three blood vessels are mediated via the 5-HT(1B) receptor, and (iii) additional mechanisms seem to be involved in coronary artery and saphenous vein contractions and middle meningeal artery relaxation following high concentrations of eletriptan. PMID- 11050305 TI - L-750355, a human beta3-adrenoceptor agonist; in vitro pharmacology and profile of activity in vivo in the rhesus monkey. AB - The profile of in vitro and in vivo biology of a human beta3-adrenoceptor agonist, (S)-N-[4-[2-[[3[(2-amino-5-pyridinyl)oxy]-2-hydroxy-propyl]amino]-eth yl]-phenyl]-4-isopropylbenzenesulfonamide, L-750355, is described. Using cloned human and rhesus beta1-, beta2- and beta3-adrenoceptors, expressed in Chinese hamster ovary (CHO) cells, L-750355 was shown to be a potent, albeit partial, agonist for the human (EC(50)=10 nM; % maximal receptor activation=49%) and rhesus (EC(50)=28 nM; % maximal receptor activation=34%) beta3-adrenoceptors. Furthermore, L-750355 stimulates lipolysis in rhesus adipocytes in vitro. L 750355 is a weak partial agonist (EC(50)=3.2 microM; % maximal receptor activation=33% ) for the human beta1-adrenoceptor but exhibits no agonist activity for rhesus beta1- or beta2-adrenoceptors of either human or rhesus origin. Administration of L-750355 to anesthetized rhesus monkeys, as a series of rising dose intravenous infusions, evokes dose-dependent glycerolemia and tachycardia with no change in mean arterial blood pressure or plasma potassium. The dose-response curve for L-750355-induced glycerolemia lies to the left of that for tachycardia. Propranolol, at a dose (0.3 mg/kg, i.v. ) that attenuates isoproterenol-induced changes in heart rate and glycerolemia, abolished L-750355 induced tachycardia but had no effect on L-750355-induced glycerolemia. PMID- 11050306 TI - Anti-muscarinic actions of mitoxantrone in isolated heart muscles of guinea pigs. AB - A hypotheses that mitoxantrone is a competitive antagonist at muscarinic cholinergic receptors was examined in guinea-pig hearts. In isolated left atrial muscle preparations, electrically paced at 2 Hz, the muscarinic agonist, carbachol, caused a concentration-dependent decrease in developed tension. Mitoxantrone caused a parallel right-ward shift of the concentration-response curve for carbachol. Schild plots for the effect of mitoxantrone on the carbachol concentration-response relationship were linear with a slope of 0.88 which was not significantly different from the unity. The right-ward shift of the carbachol concentration-response relationship by mitoxantrone significantly reversed after an additional incubation with a mitoxantrone-free solution, although the reversal was incomplete after a 2-h incubation in the mitoxantrone-free solution. Mitoxantrone caused a concentration-dependent displacement of specific [3H]quinuclidinyl benzilate binding to membrane preparations obtained from ventricular muscles of guinea-pig hearts. These results indicate that mitoxantrone acts as a competitive antagonist for the muscarinic receptors. PMID- 11050307 TI - Hypocapnic constriction in rabbit basilar artery in vitro: triggering by serotonin and dependence on endothelin-1 and alkalosis. AB - This study tested whether hypocapnic constriction of the rabbit basilar artery in vitro can be triggered by serotonin, and whether the resulting constriction is (1) due to the alkaline pH associated with hypocapnia, and (2) endothelin-1 mediated. Hypocapnic alkaline solution (25 mM NaHCO(3); pH 7.76; pCO(2) 14.2) or isocapnic alkaline solution (50 mM NaHCO(3); pH 7.73; pCO(2) 35.0) rarely altered basal tension. Serotonin (3 microM) challenge in hypocapnic or isocapnic alkaline solution resulted in near maximal tension. Washout of the serotonin did not decrease tension in 54% of the tissues, as plateau tension was maintained for 2 2.5 h. The plateau tension of washed tissues was relaxed by 1-3 microM PD145065 (Ac-D-Bhg-L-Leu-Asp-L-Ile-L-Ile-L-Trp), BQ610 (homopiperidinyl-CO-Leu-D-Trp(CHO) D-Trp), and BQ788 (N-cis-2, 6-dimethyl-piperidinocarbonyl-L-gamma-MeLeu-D-Trp (COOCH(3))-Nle), endothelin ET(A)/ET(B), endothelin ET(A), and endothelin ET(B) receptor antagonists, respectively. In contrast, serotonin-induced tension in normal solution (25 mM NaHCO(3); pH 7.42; pCO(2) 36.9) was maintained for only 40 min (mean). These results demonstrate that (1) constriction due to hypocapnia in vitro can be triggered by serotonin and is endothelin-1 mediated and (2) alkaline pH in the absence of decreased pCO(2) is sufficient to elicit the constriction triggered by serotonin. PMID- 11050308 TI - Tryptanthrin inhibits nitric oxide and prostaglandin E(2) synthesis by murine macrophages. AB - Nitric oxide (NO) and prostaglandins have been implicated in the pathogenesis of several inflammatory diseases. In this study, we investigated the effect of tryptanthrin (6,12-dihydro-6, 12-dioxoindolo-(2,1-b)-quinazoline), an antimicrobial and antitumoral plant compound isolated from Porigonum tinctorium, on NO and prostaglandin E(2) production by interferon-gamma and lipopolysaccharide-stimulated murine macrophage-like RAW 264.7 cells. Tryptanthrin markedly inhibited both NO and prostaglandin E(2) production in a dose-dependent manner. Tryptanthrin at 20 microM fully inhibited expression of inducible NO synthase, suggesting that the inhibitory effect on NO synthesis was mediated by inhibited expression of the enzyme. On the other hand, tryptanthrin had no effect on the levels of cyclooxygenase-2 protein, but inhibited cyclooxygenase enzyme activity with a ICM(50) value of 1.5 microM. Thus, tryptanthrin has the dual functions of inhibiting both NO and prostaglandin E(2) production by activated macrophages, suggesting that tryptanthrin exhibits anti inflammatory properties. PMID- 11050309 TI - Pharmacology in the noughties PMID- 11050310 TI - Physiological roles of G-protein-coupled receptor kinases revealed by gene targeting technology. PMID- 11050311 TI - Reply: receptor specificity of G-protein-coupled receptor kinases PMID- 11050312 TI - A radical approach to the pathogenesis of diabetic complications. PMID- 11050313 TI - A stressed cell? heat shock protein 70 offers protection PMID- 11050314 TI - With a little help from our (four-legged) friends PMID- 11050315 TI - Do you like your morning cup of coffee? PMID- 11050316 TI - Signalling without the receptor PMID- 11050317 TI - Dynamic mechanisms of non-classical antagonism by competitive AT(1) receptor antagonists. AB - Selective competitive angiotensin AT(1) receptor antagonists exhibit diverse patterns of antagonism of angiotensin-II-mediated responses in functional assays. These range from the classical parallel rightward shift of agonist concentration response curves with no depression of the maximum response to an apparently straightforward insurmountable antagonism with complete depression of the maximum response and no rightward shift. This article reviews some earlier equilibrium based models that have been used to explain the insurmountable antagonism, and suggests that a kinetic model might provide a more satisfactory account of the observations. Such a model might provide deeper insights into the pharmacology of G-protein-coupled receptors than the more popular equilibrium models. PMID- 11050318 TI - Nuclear receptor ligand-binding domains: three-dimensional structures, molecular interactions and pharmacological implications. AB - Nuclear receptors are members of a large family of ligand-inducible transcription factors that regulate gene programs underlying a plethora of (patho)physiological phenomena. The recent determination of the crystal structures of nuclear receptor ligand-binding domains has provided an extremely detailed insight into the intra- and intermolecular mechanisms that constitute the initial events of receptor activation and signal transduction. Here, a comprehensive mechanistic view of agonist and antagonist action will be presented. Furthermore, the novel class of partial agonists-antagonists will be described and the multiple challenges and novel perspectives for nuclear-receptor-based drug design will be discussed. PMID- 11050319 TI - Keeping in touch: sensory neurone regeneration in the CNS. AB - Adult neurones fail to regenerate when injured in the CNS, which leads to severe and irreversible functional deficits. Several important advances in understanding the reasons for this failure have been gained from the use of primary sensory neurones as a model system. The peripherally and centrally projecting branches of sensory neurones are differentially capable of regeneration, which is why these cells are ideally situated to elucidate the mechanisms that underlie regeneration failure. Such mechanisms include both a hostile environment within the spinal cord and a poor growth response following injury. For successful functional regeneration to occur, it is likely that both of these barriers will have to be surmounted, along with the challenge of guiding regrowing axons to appropriate postsynaptic targets. The contribution that the study of primary sensory neurones has made to the attainment of this goal will be reviewed. PMID- 11050320 TI - Zn(2+): a novel ionic mediator of neural injury in brain disease. AB - Zn(2+) is the second most prevalent trace element in the body and is present in particularly large concentrations in the mammalian brain. Although Zn(2+) is a cofactor for many enzymes in all tissues, a unique feature of brain Zn(2+) is its vesicular localization in presynaptic terminals, where its release is dependent on neural activity. Although the physiological significance of synaptic Zn(2+) release is little understood, it probably plays a modulatory role in synaptic transmission. Furthermore, several lines of evidence support the idea that, upon excessive synaptic Zn(2+) release, its accumulation in postsynaptic neurons contributes to the selective neuronal loss that is associated with certain acute conditions, including epilepsy and transient global ischaemia. More speculatively, Zn(2+) dis-homeostasis might also contribute to some degenerative conditions, including Alzheimer's disease. Further elucidation of the pathological actions of Zn(2+) in the brain should result in new therapeutic approaches to these conditions. PMID- 11050321 TI - Cooperative multi-modal sensing and therapeutic implications of the extracellular Ca(2+) sensing receptor. AB - The extracellular Ca(2+)-sensing receptor (CaR) is an unusual member of the diverse superfamily of seven-transmembrane domain G-protein-coupled receptors. Originally identified as the receptor providing the calciostat for extracellular ionized Ca(2+) ?[Ca(2+)](o)?, the CaR corrects small changes in [Ca(2+)](o) by regulating the secretion of the hormone that controls Ca(2+) fluxes between the blood and Ca(2+) stores in bone, and between blood and the urine. Now, research is beginning to reveal the structure and function of its unusually large N terminal head. In addition to its role as a divalent and polyvalent cation sensor, recent studies indicate that the receptor also responds sensitively to changes in ionic strength and pH. Furthermore, new work indicates that the CaR is subject to allosteric activation by L-amino acids. PMID- 11050322 TI - Analysing uncharted transcriptomes with SAGE. AB - Methods of comprehensive gene expression analysis have traditionally been limited to analysing bulk tissue or millions of cells. New modifications of serial analysis of gene expression (SAGE) have now permitted the analysis of gene expression in cell subpopulations or microanatomic structures, providing access to unexplored transcriptomes of normal and disease biology. PMID- 11050323 TI - Genome-wide analysis relating expression level with protein subcellular localization. PMID- 11050324 TI - Intron sliding in conserved gene families. PMID- 11050326 TI - Genetic studies: look no further than your own backyard PMID- 11050327 TI - Shedding light on phototaxis PMID- 11050325 TI - CMP-KDO synthetase: a plant gene borrowed from gram-negative eubacteria. PMID- 11050328 TI - The PKB that doesn't need to jump PMID- 11050329 TI - Chromosome copies and consequences PMID- 11050330 TI - Genes that fight infection: what the Drosophila genome says about animal immunity. AB - From deciphering the principles of heredity to identifying the genes that control development, the fruit fly Drosophila melanogaster is being used to deconstruct an increasing number of biological processes. Genetic studies of Drosophila responses to microbial infection have identified regulators of innate immunity that are functionally conserved in mammals. These recent findings highlight the ancient origins of animal immune responses and demonstrate the potential of Drosophila for dissecting host-pathogen interactions. The sequencing of the Drosophila genome both enhances genetic approaches and provides new clues for the identification of key components of innate immunity. This article summarizes how information gained from genomic analysis contributes to our understanding of how animals cope with infectious disease. PMID- 11050331 TI - Interplay of signaling pathways in plant disease resistance. AB - Plants are under constant threat of infection by pathogens armed with a diverse array of effector molecules to colonize their host. Plants have, in turn, evolved sophisticated detection and response systems that decipher pathogen signals and induce appropriate defenses. Genetic analysis of plant mutants impaired in mounting a resistance response to invading pathogens has uncovered a number of distinct, but interconnecting, signaling networks that are under both positive and negative control. These pathways operate, at least partly, through the action of small signaling molecules such as salicylate, jasmonate and ethylene. The interplay of signals probably allows the plant to fine-tune defense responses in both local and systemic tissue. PMID- 11050332 TI - Genomics - the new rock and roll? AB - The end of the beginning of the Human Genome Project was announced on 26 June when the working draft or first assembly was announced. Here, Ian Dunham who led the group at the Sanger Centre that produced the first complete sequence of a human chromosome reflects on how it felt to be with the genome project from the beginning. PMID- 11050334 TI - Stop press: genes that fight infections: what the drosophila genome says about animal immunity PMID- 11050333 TI - It takes two transposons to tango: transposable-element-mediated chromosomal rearrangements. AB - Transposable elements (TEs) promote various chromosomal rearrangements more efficiently, and often more specifically, than other cellular processes(1-3). One explanation of such events is homologous recombination between multiple copies of a TE present in a genome. Although this does occur, strong evidence from a number of TE systems in bacteria, plants and animals suggests that another mechanism - alternative transposition - induces a large proportion of TE-associated chromosomal rearrangements. This paper reviews evidence for alternative transposition from a number of unrelated but structurally similar TEs. The similarities between alternative transposition and V(D)J recombination are also discussed, as is the use of alternative transposition as a genetic tool. PMID- 11050336 TI - Are you interested in writing short headline articles? PMID- 11050337 TI - Cut and run: the dramatic rise of transnational logging in the tropics. PMID- 11050335 TI - Internal ribosome initiation of translation and the control of cell death. AB - The majority of cellular stresses lead to the inhibition of cap-dependent translation. Some mRNAs, however, are translated by a cap-independent mechanism, mediated by ribosome binding to internal ribosome entry site (IRES) elements located in the 5' untranslated region. Interestingly, IRES elements are found in the mRNAs of several survival factors, oncogenes and proteins crucially involved in the control of apoptosis. These mRNAs are translated under a variety of stress conditions, including hypoxia, serum deprivation, irradiation and apoptosis. Thus, IRES-mediated translational control might have evolved to regulate cellular responses in acute but transient stress conditions that would otherwise lead to cell death. PMID- 11050338 TI - No question: seed dispersal matters. PMID- 11050339 TI - Female secondary sexual characteristics: appearances might be deceptive. PMID- 11050341 TI - Trophic cascades in terrestrial ecosystems. Reflections on Polis et al. PMID- 11050340 TI - What enables trophic cascades? Commentary on Polis et al. PMID- 11050342 TI - Communication between plants: this time it's real. PMID- 11050343 TI - A not-quite-so inordinate fondness for beetles. PMID- 11050344 TI - Erratum. PMID- 11050345 TI - Old British grasslands might die hard in the face of climate change. PMID- 11050346 TI - Foreshadowing the Cretaceous-Tertiary extinction: none if by land, two if by sea. PMID- 11050347 TI - Seasonality in coastal benthic ecosystems. AB - For historical reasons, knowledge about seasonality in the dynamics of marine benthic suspension feeders from temperate areas comes mainly from studies of cold temperate seas. Recent surveys of Mediterranean taxa show different patterns from those observed in cold temperate seas, which are characterized by winter dormancy. In the Mediterranean, summer dormancy predominates among taxa and appears to be related to energetic constraints. Temperature and food availability are crucial to the dynamics of benthic suspension feeders. However, because these factors tend to be positively correlated in cold temperate seas, it is difficult to distinguish between their effects. Such correlation does not occur in Mediterranean ecosystems. The contrast between recent studies in the Mediterranean and in other areas can help to disentangle confounded environmental controls. PMID- 11050348 TI - Rare genomic changes as a tool for phylogenetics. AB - DNA sequence data have offered valuable insights into the relationships between living organisms. However, most phylogenetic analyses of DNA sequences rely primarily on single nucleotide substitutions, which might not be perfect phylogenetic markers. Rare genomic changes (RGCs), such as intron indels, retroposon integrations, signature sequences, mitochondrial and chloroplast gene order changes, gene duplications and genetic code changes, provide a suite of complementary markers with enormous potential for molecular systematics. Recent exploitation of RGCs has already started to yield exciting phylogenetic information. PMID- 11050349 TI - Population variability in space and time. AB - One of the most ubiquitous phenomena of all natural populations is their variability in numbers in space and time. However, there are notable differences among populations in the way the population size fluctuates. One of the major challenges in population and community ecology is to explain and understand this variety and to find possible underlying rules that might be modified from case-to case. Population variability also has a spatial component because fluctuations are often synchronized over relatively large distances. Recently, this has led to growing interest in how 'internal' (density-dependent) processes interact with 'external' factors such as environmental variability. PMID- 11050350 TI - Behavioral endocrinology of mammalian fatherhood. AB - Mammalian fatherhood involves a muted version of the maternal experience. In spite of previous assumptions to the contrary, hormones influence mammalian paternal behavior. Naturally paternal males experience dynamic changes in the same hormones involved in maternal behavior and these hormones have access to the same brain pathways. Men becoming fathers for the first time are similar to their female partners too. These recent studies are still correlational, but promise to illuminate maternal behavior and to biologically validate the experiences of involved fathers. PMID- 11050351 TI - When is a trophic cascade a trophic cascade? PMID- 11050352 TI - The effects of motion on trunk biomechanics. AB - OBJECTIVE: To review the literature that evaluates the influence of trunk motion on trunk strength and structural loading. BACKGROUND: In recent years, trunk dynamics have been identified as potential risk factors for developing low-back disorders. Consequently, a better understanding of the underlying mechanisms involved in trunk motion is needed. METHODS: This review summarizes the results of 53 studies that have evaluated trunk motion and its impact on several biomechanical outcome measures. The biomechanical measures consisted of trunk strength, intra-abdominal pressure, muscle activity, imposed trunk moments, and spinal loads. Each of these biomechanical measures was discussed in relation to the existing knowledge within each plane of motion (extension, flexion, lateral flexion, twisting, and asymmetric extension). RESULTS: Trunk strength was drastically reduced as dynamic motion increased, and males were impacted more than females. Intra-abdominal pressure seemed to only be affected by trunk dynamics at high levels of force. Trunk moments were found to increase monotonically with increased trunk motion. Both agonistic and antagonistic muscle activities were greater as dynamic characteristics increased. As a result, the three-dimensional spinal loads increase significantly for dynamic exertions as compared to isometric conditions. CONCLUSIONS: Trunk motion has a dramatic affect on the muscle coactivity, which seems to be the underlying source for the decrease strength capability as well as the increased muscle force, IAP, and spinal loads. This review suggests that the ability of the individual to perform a task "safely" might be significantly compromised by the muscle coactivity that accompanies dynamic exertions. It is also important to consider various workplace and individual factors when attempting to reduce the impact of trunk motions during dynamic exertions. Relevance This review provides insight as to why trunk motions are important risk factors to consider when attempting to control low back disorders in the workplace. It is apparent that trunk motion increases the risk of low-back disorders. To better control low-back disorders in industry, more comprehensive knowledge about the impact of trunk motion is needed. A better understanding of muscle coactivity may ultimately lead to reducing the risk associated with dynamic exertions. PMID- 11050353 TI - Load-bearing and stress analysis of the human spine under a novel wrapping compression loading. AB - OBJECTIVE: To examine biomechanics of the human spine under a novel compression loading that follows the curvature of the spine.Design. The detailed response of the spine is predicted and compared under various types of compression loading at different postures. BACKGROUND: The posture and loading configuration could be so adjusted as to increase load-bearing capacity and stability of the spine in compression while minimizing the muscle activity and risk of tissue injury. METHODS: The nonlinear finite element formulation of wrapping elements sliding over solid body edges is developed and used to study the load-bearing capacity of simplified beam-rigid body thoracolumbar (T1-S1) and lumbosacral (L1-S1) spines under a wrapping compression force. The load-bearing and stress analysis of a detailed model of the lumbar spine, L1-S1, is also investigated under five wrapping loads resulting in differential compression forces at various levels. Follower load at L1, axially fixed compression at L1, and combined axially fixed compression and moments load are also considered for comparison. For the detailed model, the effect of changes in the position of wrapping elements and in the lumbar curvature on results are considered. RESULTS: The idealized wrapping loading stiffens the spine, allowing it to carry very large compression loads without hypermobility. It diminishes local segmental shear forces and moments as well as tissue stresses. CONCLUSIONS: In comparison to fixed axial compression, the compression loading by wrapping elements that follow the spinal curvatures increases the load-bearing capacity in compression and provides a greater margin of safety against both instability and tissue injury. Relevance These findings suggest a plausible mechanism in which postural changes and muscle activation patterns could be exploited to yield a loading configuration somewhat similar to that of the wrapping loading, i.e., the net reaction force at various levels passes through discs nearly normal to their mid-height plane. To alleviate hypermobility in compression, the wrapping loading could also allow for the application of meaningful compression loads in experimental as well as model studies of the multi-segmental spinal biomechanics. PMID- 11050354 TI - Practice-related changes in lumbar loading during rapid voluntary pulls made while standing. AB - OBJECTIVE: To determine if five days of practice on a novel dynamic, multi-joint pulling task resulted in lower magnitudes of lumbar loading or a more consistent relationship between pulling force and lumbar loading. DESIGN: A repeated measures design compared how practice influenced the magnitude of lumbar torque and the correlations between lumbar torque and pulling force. BACKGROUND: Previous studies suggest that practice can decrease the magnitude of lumbar loading on simple manual material handling tasks, but it is unknown whether practice reduces lumbar loading for more complex tasks. Neither is it known whether the consistency of lumbar loading increases with practice. METHODS: Ten healthy adults practiced impulse-like horizontal pulls to targets equaling 20%, 40% and 80% of their estimated maximal dynamic pulling force over 5 days. Movements were unrestrained, other than keeping the feet flat on the ground. We used a four-segment, sagittal plane inverse dynamics model to compute lumbar, hip, knee, and ankle torques on days 1 and 5 from ground reaction forces and moments, pulling forces, and kinematics. RESULTS: An analysis of variance showed significant practice-related changes in lumbar torque at the time of peak pulling force (lumbar torque(peakPF)). The lumbar torque(peakPF) decreased for the 20% pulls, did not change for the 40% pulls, and increased for the 80% pulls. Two subjects showed a significant decrease in lumbar torque(peakPF) for all three force levels. Coefficients of determination between pulling force and lumbar torque (r(2)(PF,LT): a measure of the consistency of the relationship between these two variables) were significantly higher on day 5 than day 1. CONCLUSIONS: Practice on a novel pulling task changed the magnitude of lumbar torques and increased their correlation with pulling force, suggesting that subjects learned strategies that improve motor control of lumbar torques. Relevance The study showed that the magnitude and consistency of lumbar loading changed spontaneously as subjects practiced a novel multijoint pulling task. Such changes may decrease the risk of low-back injury. PMID- 11050355 TI - Glenohumeral kinematics and capsulo-ligamentous strain resulting from laxity exams. AB - OBJECTIVE: Identification and quantification of strain in shoulder capsular ligamentous structures during clinical exams and validation of this testing on cadavers. METHODS: Mercury strain gauges were sutured in seven locations on shoulders from cadavers. An electromagnetic tracker quantified humeral head translations during laxity exams. Strain and humeral position were acquired during performance of Sulcus, Feagin, Apprehension, Load and Shift, Drawer, and Hawkins tests. RESULTS: Anterior humeral head translation in neutral position was primarily constrained by the coracohumeral ligament. With the arm abducted, anterior middle and inferior ligaments also became active. External rotation and abduction activated inferior and middle capsules. Posterior capsule constrained motion for posterior tests in neutral and abduction. Superior and inferior capsular ligaments were active during inferior tests in neutral position. With abduction, inferior ligaments provided primary translation constraint. CONCLUSION: Study of kinematics and strain evaluation on cadavers can yield useful information on mechanisms of glenohumeral instability. Relevance This study clarifies the contribution of specific structures of the shoulder to strain in the joint capsule. It also identifies which structures are challenged by provocative laxity exams commonly used by orthopaedic physicians. PMID- 11050356 TI - Joint angle-dependence of elbow flexor activation levels during isometric and isokinetic maximum voluntary contractions. AB - OBJECTIVE: The influences of elbow joint angle and the type of contraction on the activation levels of biceps brachii and brachioradialis during maximum voluntary isometric and isokinetic contractions were investigated. DESIGN: A within-session repeated measures design. BACKGROUND: Activation of synergistic elbow flexor muscles has been reported to be affected disparately by elbow joint angle and contraction type. METHODS: Ten subjects performed concentric isokinetic, eccentric isokinetic, and isometric maximum voluntary contractions of the elbow flexor muscles. For the isokinetic contractions the activation levels of two ranges of motion were compared. For the isometric contractions the activation levels at two joint angles were compared. The activation levels of the biceps brachii and brachioradialis acquired simultaneously using bipolar surface electrodes and a surface electrode array were compared. RESULTS: Results from the electrode array were similar to those acquired using conventional bipolar electrodes. The activation of biceps brachii was significantly affected by joint angle during concentric isokinetic and isometric maximum voluntary contractions. The activation of brachioradialis was significantly affected by joint angle only during eccentric isokinetic maximum voluntary contractions. CONCLUSIONS: The results confirm that joint angle and contraction type contribute to the distinction between the activation of synergistic elbow flexor muscles during isometric and isokinetic contractions. Relevance The results point to the complexity of control of elbow joint synergists and raise questions about the plasticity of this dependency of elbow flexor activation on joint angle. Solutions to these questions are of importance in the areas of upper extremity rehabilitation and modeling the upper extremity neuromechanics. PMID- 11050357 TI - Initial stability of fully and partially cemented femoral stems. AB - OBJECTIVE: To test the initial stability of a newly designed partially cemented femoral stem in comparison with a fully cemented conventional stem. DESIGN: An in vitro study to determine the interface motion between femoral stem and bone as a response to loading. BACKGROUND: The aim of the new prosthesis design is a proximal load transfer by a defined partial cement fixation in the proximal femur region and a slim prosthesis stem in the distal region. Before a clinical study can be started, the new stem has to show an initial stability comparable to that of fully cemented prostheses. METHOD: Six paired fresh cadaveric femora were used for the testing of the new partially cemented stem (Option 3000, Mathys Orthopaedics, Bettlach, Switzerland) and a fully cemented stem (Weber Shaft, AlloPro, Baar, Swizerland). Under cyclic loading up to 1600 N hip joint forces, the interface motion between implants and bone was measured at six locations. RESULTS: Both stems showed uncritical interface motions below 43 microm. However, the Option 3000 stem exhibited significantly smaller motions in the proximal region and slightly larger movements in the distal regions than the Weber prosthesis. CONCLUSIONS: The new type of partially cemented stem provided a comparable initial stability to the fully cemented Weber prosthesis. Relevance The high initial stability of the Option 3000 stem justified the clinical use of the new implant. More than 100 implantations in the last three years, with very good preliminary clinical results, support the preclinical findings. PMID- 11050358 TI - Biomechanical study on the effect of twisted human patellar tendon. AB - OBJECTIVE: To compare the stiffness and maximum strength between the untwisted and twisted free-tendon. DESIGN: 22 twisted and untwisted sectioned-specimens of human cadaver patellar tendons were used and pulled to failure to obtain load deformation profiles from which stiffness, maximum load to failure and elastic elongation limit were derived. BACKGROUND: In the reconstruction of the deficient anterior cruciate ligament, the use of the central one-third of the patellar tendon is a well-established procedure in which, prior to insertion, the tendon graft may be twisted to mimic the natural orientation of the anterior cruciate ligament in the knee joint. RESULTS: The untwisted tendons had a mean stiffness of 36.5 kg/mm (SD, 16.6 kg/mm) and maximum load of 165.9 kg (SD, 86.8 kg). With a 90 degrees twist, the average stiffness of the twisted tendon was 66.5 kg/mm (SD, 25.4 kg/mm), with maximum load at 364.5 kg (SD, 109.9 kg), an increase of over 100%. The elastic elongation limit, or allowable elongation before permanent deformation or failure, was significantly larger in twisted tendons by 35%. CONCLUSION: Twisting increased the resistance to deformation of the tendon in this study. Relevance The finding supports the surgical practice of pre-twisting tendon grafts for anterior cruciate ligament reconstruction, based on the premise that a stronger and stiffer graft provides a more favourable outcome. PMID- 11050359 TI - Doppler imaging of vibrations as a tool for quantifying first tarsometatarsal joint stiffness. AB - OBJECTIVE: To assess whether first tarsometatarsal joint stiffness can be measured by Doppler imaging of vibrations and if so, to assess reference values. DESIGN: Repeated in vivo Doppler imaging of vibrations measurements at the first tarsometatarsal joint in healthy persons. BACKGROUND: Clinical hypermobility of the first tarsometatarsal joint is an important factor in a hallux valgus deformity. No objective and non-invasive test is available to quantify first tarsometatarsal joint mobility. Doppler imaging of vibrations, a technique recently developed to measure joint stiffness, might be an effective tool to quantify stiffness of this joint. METHODS: Vibrations were applied to the head of the first metatarsal in 46 feet of 23 healthy subjects and picked up by a transducer at both sides of the first tarsometatarsal joint. A pilot study was performed on three patients with hypermobility of the first tarsometatarsal joint. Measurements are expressed in threshold units related to colour Doppler imaging. RESULTS: The values of the threshold units were found to be very similar in healthy persons, with a good repeatibility; 95% of the healthy persons had a threshold unit below 3.4. No significant difference was found between the left and right foot, or between male and female subjects. Also there was no significant correlation with age or weight of the subjects. In the three patients with first tarsometatarsal hypermobility we found threshold units above 5. CONCLUSIONS: With Doppler imaging of vibrations first tarsometatarsal joint stiffness can be measured in healthy persons in a non-invasive and objective way. In a pilot study, three patients with first tarsometatarsal hypermobility showed lower stiffness values than the healthy subjects. Relevance This study presents a new method for quantification of first tarsometatarsal joint stiffness and provides reference values in healthy persons. First measurements on patients gave promising results to future use of this method for assessment of clinical hypermobility in hallux valgus patients. PMID- 11050360 TI - A computer analysis of reflex eyelid motion in normal subjects and in facial neuropathy. AB - OBJECTIVE: To demonstrate how computerized eyelid motion analysis can quantify the human reflex blink. DESIGN: Seventeen normal subjects and 10 patients with unilateral facial nerve paralysis were analyzed. BACKGROUND: Eyelid closure is currently evaluated by systems primarily designed to assess lower/midfacial movements. The methods are subjective, difficult to reproduce, and measure only volitional closure. Reflex closure is responsible for eye hydration, and its evaluation demands dynamic analysis. METHODS: A 60Hz video camera incorporated into a helmet was used to analyze blinking. Reflective markers on the forehead and eyelids allowed for the dynamic measurement of the reflex blink. Eyelid displacement, velocity and acceleration were calculated. The degree of synchrony between bilateral blinks was also determined. RESULTS: This study demonstrates that video motion analysis can describe normal and altered eyelid motions in a quantifiable manner. CONCLUSIONS: To our knowledge, this is the first study to measure dynamic reflex blinks. Eyelid closure may now be evaluated in kinematic terms. This technique could increase understanding of eyelid motion and permit more accurate evaluation of eyelid function. Dynamic eyelid evaluation has immediate applications in the treatment of facial palsy affecting the reflex blink. Relevance No method has been developed that objectively quantifies dynamic eyelid closure. Methods currently in use evaluate only volitional eyelid closure, and are based on direct and indirect observer assessments. These methods are subjective and are incapable of analyzing dynamic eyelid movements, which are critical to maintenance of corneal hydration and comfort. A system that quantifies eyelid kinematics can provide a functional analysis of blink disorders and an objective evaluation of their treatment(s). PMID- 11050361 TI - Measuring spinal motion in rowers: the use of an electromagnetic device. AB - OBJECTIVE: To determine whether a skin-mounted electromagnetic measurement device can be used to measure lumbar spinal motion in rowers and discriminate between variations in technique. DESIGN: The lumbar spinal kinematics of elite level rowers were assessed with an electromagnetic device (Flock of Birds) during ergometer training using five technique variants. The system was correlated with sagittal MRI imaging of the lumbar spine and pelvis. BACKGROUND: Rowing technique is related to performance and injury. This study sought to test a method to obtain quantitative data on the effect of changes in technique on lumbar spinal and pelvic motion. METHODS: Lumbar spinal and pelvic motion were measured in six elite level rowers on an ergometer using normal rowing technique, three common bad technique variants, and after a 10 min piece of rowing simulating fatigue. RESULTS: Significant differences were found between the different rowing techniques for femoral, thoraco-lumbar and lumbo-sacral flexion. CONCLUSIONS: Kinematic parameters of spinal motion of rowers can be measured dynamically and used to quantify and discriminate between good and bad rowing styles. Relevance There is a consistent style of "good" rowing technique on an ergometer. Deviations from this technique can be measured and this will allow studies to be conducted comparing rowing styles between rowers of different standards and between different boat houses. There is potential to develop a dynamic feedback system to the rower that will assist in training and eradicate bad technique variants. PMID- 11050362 TI - Changes in lumbar lordosis modify the role of the extensor muscles. AB - STUDY DESIGN: Fiber angles of longissimus thoracis and iliocostalis lumborum at L3 were documented in vivo, using high resolution ultrasound, with the lumbar spine in neutral curve and when fully flexed. OBJECTIVES: To evaluate the effect of changes in lumbar curvature on the mechanics of these muscles. BACKGROUND: Full flexion modifies the failure tolerance of the lumbar spine, determines the load distribution among muscle and passive tissues, and modulates the types of tissue damage that occur. Related to this issue are the possible changes in muscle line of action with full flexion which changes the ability of the spine to support shear loads. METHODS: Nine normal men and 5 normal women were scanned in three positions: (1) an upright standing posture; (2) with the hips flexed to approximately 30 degrees and the spine fully flexed; (3) hips flexed but the spine returned to a neutral curvature. RESULTS: Mean longissimus/iliocostalis fiber angles for upright standing, hips flexed-spine flexed, and hips flexed spine neutral lordosis were 25. 7 degrees, 10.7 degrees and 28.3 degrees, respectively. CONCLUSIONS: Anterior shear load on the lumbar spine has been recently shown to be highly related to the risk of reporting a back injury. Bending forward allowing the spine to fully flex changes the line of action of the largest lumbar extensor muscles compromising their role to support anterior shear forces. Relevance Fiber angles of longissimus thoracis and iliocostalis lumborum were documented with high resolution ultrasound at L3, with the spine in neutral curvature and fully flexed. Full lumbar flexion changes the line of action of these muscle compromising their role to support anterior shear forces on the spine - anterior shear forces have been recently documented to be highly related to the risk of reporting a back injury. PMID- 11050363 TI - A comparison of vertical force and temporal parameters produced by an in-shoe pressure measuring system and a force platform. AB - OBJECTIVE: To investigate the ability of Pedar in-shoe system to measure vertical force accurately, by comparing it with the Kistler force platform. DESIGN: In vivo experiment in normal subjects. BACKGROUND: It has been suggested Pedar is highly reliable, but absolute accuracy of the system with regard to force measurement has not been comprehensively tested. METHODS: Sampling at 99 Hz, using five healthy subjects, simultaneous data were collected barefoot, and in three types of shoes (Trainers, Oxfords, Slip-on Deck Type). Six variables obtained from the force/time curve from each footstep were compared. RESULTS: The temporal data recorded by Pedar correlated well with that obtained using Kistler, with significant differences only in overall duration of the step in Deck shoes (P<0. 001) and Oxford shoes (P<0.01), and peak to peak barefoot (P<0.01). Pedar recorded a lower first peak force and mid-peak force in all cases (P<0.001). However, the magnitude of the 2nd peak force recorded by both systems was significantly different only in Trainers (P<0.05) and Oxford shoes (P<0.001). The impulse data obtained with Oxford shoes was not significantly different, although barefoot, Trainers and Deck shoes were significantly lower (P<0.001) for Pedar. CONCLUSIONS: In most cases, comparison of data recorded by the two systems provided good evidence for the accuracy and reliability of temporal measurements and second peak force measurements taken with the Pedar in-shoe system. Relevance In-shoe pressure data provides evidence for clinical decisions if the systems utilised are proven to be valid, repeatable and accurate. Comparison with an established force platform enables some assessment of these factors. PMID- 11050364 TI - Technique to measure lumbar curvature in the ergonomic evaluation of chairs: description and validation. AB - OBJECTIVE: To describe and validate a system to record the lumbar curvature, specially designed for the ergonomic evaluation of chairs. DESIGN: The technique consists of a flexible goniometer and requires a model of lumbar flexion (relationship between length and angle). BACKGROUND: Most of the techniques do not allow using the backrest of the chair or do not record continuously. METHODS: Precision, repeatability and reproducibility of the measurements have been analysed, as well as the lumbar flexion model. RESULTS: A suitable lumbar model of flexion and the repeatability and reproducibility errors obtained are presented. CONCLUSIONS: The linear model is a good one to represent the flexion of the lumbar spine. The measurements are reproducible and the errors are similar or lower than with conventional inclinometers. Relevance The device may help to investigate the relationship between lumbar pain and curvature, and to evaluate the lumbar curvature in sedentary work. PMID- 11050365 TI - Learning and memory in pain pathways. PMID- 11050366 TI - Pindolol, a beta-adrenoceptor blocker/5-hydroxytryptamine(1A/1B) antagonist, enhances the analgesic effect of tramadol. AB - The ability of pindolol, a beta-adrenoceptor blocker/5-hydroxytryptamine(1A/1B) antagonist, to enhance the clinical antidepressant response to selective serotonin re-uptake inhibitors is generally attributed to a blocking of the feedback that inhibits the serotoninergic neuronal activity mediated by somatodendritic 5-hydroxytryptamine (5-HT)(1A) autoreceptors. The current study examined the ability of pindolol to enhance the analgesic effect of tramadol, an atypical centrally-acting analgesic agent with relatively weak opioid receptor affinity and which, like some antidepressants, is able to inhibit the re-uptake of 5-HT in the raphe nuclei. Racemic pindolol (2 mg/kg, s.c.), rendered analgesic a non-effective acute dose of tramadol (10-40 mg/kg, i.p.) in two nociceptive tests: a hot plate test in mice and a plantar test in rats. Moreover, (+/-)8-OH DPAT (0.125-1 mg/kg, s.c.), a selective 5-HT(1A) agonist, reduces the analgesic effect of tramadol in the same tests. These results suggest an implication of the somatodendritic 5-HT(1A) receptors in the analgesic effect of tramadol and open a new adjuvant analgesic strategy for the use of this compound. PMID- 11050367 TI - Capsaicin evoked pain and allodynia in post-herpetic neuralgia. AB - The hypothesis that the pain and allodynia associated with post-herpetic neuralgia (PHN) is maintained by a combination of input from preserved primary afferent nociceptors and sensitization of central pain transmitting neurons was examined in 17 subjects with PHN. Pain, allodynia, thermal sensory function, cutaneous innervation, and response to controlled application of 0.075% capsaicin were measured. Compared to mirror-image skin, applying capsaicin on a 9 cm(2) area of PHN skin significantly increased overall PHN pain and allodynia in 11 of 17 subjects. These 'capsaicin responders' were characterized by higher average daily pain, higher allodynia ratings, and relatively preserved sensory function at baseline compared to the non-responders. In three of the 'capsaicin responders' the area of allodynia expanded into previously non-allodynic and non painful skin that had normal sensory function and cutaneous innervation. These observations support the hypothesis that allodynia in some PHN patients is a form of chronic secondary hyperalgesia maintained by input from intact and possibly 'irritable' primary afferent nociceptors to a sensitized CNS. PMID- 11050368 TI - Assessing the significant others of chronic pain patients: the psychometric properties of significant other questionnaires. AB - Contemporary reviews of psychological models of chronic pain have favoured behavioural and cognitive-behavioural formulations. These have often assumed that pain behaviours are maintained by environmental reinforcers. One of the most commonly hypothesized sources of reinforcement has been patients' significant others. Further, it has often been recognized that significant others may also be affected by pain behaviours and that they may experience changes in their lifesyles and in their mood as a consequence of living with someone who has pain. Somewhat surprisingly, relatively little clinical research has been published investigating significant others and their relationships with pain patients. Among other things, one of the limiting factors has been the lack of measurement tools available for assessing the relevant variables thought to be important with regards to significant others (such as their responses to, and perceptions of, chronic pain). This study attempted to remedy this situation by developing and testing the psychometric properties of a number of questionnaires specifically designed for significant others of chronic pain patients. The questionnaires have been selected to assess both significant others' (behavioural and cognitive) responses to pain as well as the extent to which pain impacts on their lives. Although not all of the questionnaires were found to possess equally strong psychometric properties, the availability of several solid measures opens the way for more empirical analyses of significant others and their interactions with chronic pain patients. PMID- 11050369 TI - A randomized trial of a cognitive-behavioral program for enhancing back pain self care in a primary care setting. AB - Back pain is a significant health care problem that has been managed unsatisfactorily in primary care settings. Providers typically address medical issues but do not adequately address patient concerns or functional limitations related to back pain. We evaluated a brief intervention for primary care back pain patients designed to provide accurate information about back pain, instill attitudes favorable towards self care, reduce fears and worries, assist patients in developing personalized action plans to manage their back pain, and improve functional outcomes. Patients enrolled in a large health maintenance organization were invited to participate in an educational program to improve back pain self care skills 6-8 weeks after a primary care back pain visit. Patients (n=226) were randomly assigned to a Self Care intervention or to Usual Care, and were assessed at baseline, 3-, 6-, and 12-months. The intervention involved a two-session Self Care group and an individual meeting and telephone conversation with the group leader, a psychologist experienced in chronic pain management. The intervention was supplemented by educational materials (book and videos) supporting active management of back pain. The control group received usual care supplemented by a book on back pain care. Participants assigned to the Self Care intervention showed significantly greater reductions in back-related worry and fear-avoidance beliefs than the control group. Modest, but statistically significant, effects on pain ratings and interference with activities were also observed. PMID- 11050370 TI - Effects of morphine on immune response in rats with sciatic constriction injury. AB - We investigated the effects of acute and of chronic morphine treatment on T lymphocyte function and natural killer (NK) cell activity in rats receiving chronic constriction injury (CCI) of the sciatic nerve. T-Lymphocyte function was evaluated based on concanavalin-A (ConA)- and phytohemagglutinin (PHA)-induced splenocyte proliferation. The effects of morphine on thermal hyperalgesia were also assessed by measuring paw withdrawal latency (PWL) in rats. All of the rats that received CCI developed thermal hyperalgesia while sham-operated rats did not. Thermal hyperalgesia was dose-dependently reversed after acute (single injection) and after chronic (daily injection for 7 days) administration of morphine but persisted in saline-treated CCI rats. There was no significant difference between sham and saline-treated CCI groups in splenocyte proliferation and NK cell activity. NK cell activity and splenocyte proliferation induced by ConA and PHA were significantly suppressed by acute morphine treatment in a dose dependent manner. The reversal of the thermal hyperalgesia persisted throughout the period of chronic morphine treatment. No tolerance to the suppression of NK cell activity and splenocyte proliferation was observed after chronic morphine treatment. These data suggest that both acute and chronic morphine treatment can cause a dose-dependent reversal of thermal hyperalgesia and inhibition of NK cell activity and splenocyte proliferation in rats with sciatic CCI, without concomitant development of tolerance. Opioid therapy for chronic neuropathic pain should be used cautiously, especially in immune-compromised cases. PMID- 11050371 TI - Intradermal injection of norepinephrine evokes pain in patients with sympathetically maintained pain. AB - Tissue injuries, with or without involvement of nerves, may lead to ongoing pain and hyperalgesia to external stimuli. In a subset of patients, the pain is maintained by sympathetic efferent activity (SMP). We investigated if the peripheral administration of the alpha-adrenergic agonist, norepinephrine (NE), in physiologically relevant doses resulted in pain in patients with SMP. To establish the dose of intradermal NE required to induce cutaneous vasoconstriction, NE (1 nM-10 microM, 30 microl) was injected under a laser Doppler probe on the volar forearm of seven normal subjects. A decrease in blood flow was evident at a dose of 10 microM. Twelve patients (five male, seven female) diagnosed to have SMP based on the decrease in pain by a local anesthetic sympathetic blockade (70+/-6%) were enrolled in the study. Pain ratings were obtained continuously for 5 min after intradermal injections of saline and NE (0.1-10 microM) into their hyperalgesic zone and the mirror-image contralateral side. Injections were done during the period of pain relief following a local anesthetic sympathetic blockade. Similar injections were made in eight control subjects. On the affected side of the patients, the two highest concentrations of NE (1 and 10 microM) caused significantly more pain than saline (P<0.05, ANOVA). In contrast, there was no significant pain induced by the NE injections in the unaffected side and in control subjects. Six of nine patients tested reported a marked decrease in pain and hyperalgesia following infusion of phentolamine (1 mg/kg over 10 min). Two of the three patients who did not receive pain relief following phentolamine infusion also did not report pain to the NE injections. We conclude that NE injections produce pain in SMP patients at doses that are at the threshold for producing vasoconstriction. These studies support a role for cutaneous adrenoceptors in the mechanisms of sympathetically maintained pain. PMID- 11050372 TI - The anterior pretectal nucleus participates as a relay station in the glutamate-, but not morphine-induced antinociception from the dorsal raphe nucleus in rats. AB - The anterior pretectal nucleus (APtN) and the dorsal raphe nucleus (DRN) are involved in descending pathways that control noxious inputs to the spinal cord and participate in the normal physiological response to noxious stimulation. Evidence has also been provided for the involvement of the APtN acting as a relay station through which the DRN partly modulates spinal nociceptive messages. In the present study, the effects of microinjecting glutamate or morphine into the DRN on the latency for the tail withdrawal reflex after noxious heating of the skin were examined in rats in which hyperbaric lidocaine (5%), naloxone (a non selective opioid antagonist) or methiothepin (a non-selective 5-HT(1) antagonist) was previously microinjected into the APtN. Microinjection of glutamate (38 nmol/0.25 microl) into the DRN evoked strong but short-lasting antinociception that was fully inhibited by the previous administration of lidocaine (0.25 microl), naloxone (2.7 nmol/0.25 microl), or methiothepin (1 nmol/0.25 microl). A smaller dose of methiothepin (0.5 nmol/0.25 microl) significantly reduced the effect of glutamate. Microinjection of morphine (7.5 nmol/0.25 microl) into the DRN evoked strong and long-lasting antinociception that was not significantly changed by previous microinjection of lidocaine into the APtN. These results confirm that APtN integrity is at least in part necessary for the antinociceptive effects of stimulating the DRN, and that at least opioid and 5-HT1 mechanisms in the APtN participate as neuromodulators in the DRN-APtN connection. The results demonstrate that the antinociceptive effects of stimulating the DRN-APtN path depend on the activation of cell bodies in the DRN that can be excited by the local administration of glutamate, but not morphine. The study also further supports the notion that the DRN is involved in both descending and ascending pain inhibitory systems. PMID- 11050373 TI - Numerous adrenal chromaffin cell preparations fail to produce analgesic effects in the formalin test or in tests of acute pain even with nicotine stimulation. AB - Previous studies have reported that intrathecal implants of a variety of adrenal chromaffin cell preparations all produce analgesic effects in rodents. The major objective of the present study was to determine if any adrenal chromaffin cell preparations produce more robust analgesic effects than other cell preparations. The present study included adult rat adrenal chromaffin tissue allografts, purified adult bovine chromaffin cells, and polymer-encapsulated calf adrenal chromaffin cells, all prepared according to previously published procedures, as well as purified calf adrenal chromaffin cells. Previous studies have also suggested that immunosuppression may play a role in graft survival, and potentially increase the magnitude of analgesic effects, so the present study included both immunosuppressed and non-immunosuppressed groups (cyclosporin A, 10 mg/kg per day). Behavioral tests included the formalin test; and a dorsal tail flick, hot-plate, and paw-pinch test, conducted sequentially 2 min after systemic nicotine (0.1 mg/kg) to evoke release from the chromaffin cells, as previously reported. Analgesic effects related to morphine and nicotine were detected, and consistent differences in performance could be detected between individual animals. Surprisingly, no analgesic effects were detectable with any of the four chromaffin cell preparations, with or without immunosuppression, in the formalin test or with nicotine stimulation in tests of acute pain. PMID- 11050374 TI - Effect of tonic muscle pain on short-latency jaw-stretch reflexes in humans. AB - The modulation of human jaw-stretch reflexes by experimental muscle pain was studied in three experiments. Short-latency reflex responses were evoked in the masseter and temporalis muscles by fast stretches (1 mm displacement, 10 ms ramp time) before, during and 15 min after a period with tonic pain. In Expt. I, a dose of 5.8% hypertonic or 0.9% isotonic (control) saline was infused in random order into the left masseter for up to 15 min (n=12). The level of excitation of the left masseter was kept constant at 15% of maximal effort by visual feedback and on-line calculation of the root-mean-square value of the surface electromyogram (sEMG). In Expt. II, a dose of 5.8% saline was infused into the left masseter but with feedback from the right masseter sEMG (n=12). In Expt. III, both sEMG and intramuscular (im) EMG was recorded from the left and right masseter muscles. The feedback was from either the sEMG or imEMG of the left masseter in which 5.8% saline was infused (n=12). In all experiments, subjects continuously rated their perceived pain intensity on a 10-cm visual analogue scale (VAS). Infusion of 5.8% saline caused moderate levels of pain (mean VAS 4.9 5.0 cm) whereas infusion of 0.9% saline was almost pain-free (mean VAS 0.3 cm). The pre-stimulus EMG activity in the masseter, which served as the feedback muscle during the recording, was constant across the different conditions. During painful infusion of 5.8% saline in Expts. I and III, the pre-stimulus sEMG activity in the non-painful masseter was significantly higher than baseline when the sEMG on the painful side was used as feedback signal, and in Expt. II significantly lower on the painful side when the non-painful side served as feedback signal (Student-Newman-Keuls: P<0.05). Isotonic saline did not affect the pre-stimulus sEMG activity or the jaw-stretch reflex parameters. The peak-to peak amplitude of the stretch reflex in the painful masseter normalized to the pre-stimulus EMG activity was significantly higher during the pain conditions compared with the pre- and post-infusion conditions in all experiments. These results indicate that experimental jaw-muscle pain facilitates the short-latency (8-9 ms), probably monosynaptic, jaw-stretch reflex as revealed by both sEMG and imEMG. This effect could not be accounted for by variability in pre-stimulus EMG activity. An increased sensitivity of the fusimotor system at this level of static muscle excitation is suggested as a possible mechanism, which could contribute to an increased stiffness of the jaw-muscles during pain. PMID- 11050375 TI - Pressure pain threshold and needle acupuncture in chronic tension-type headache- a double-blind placebo-controlled study. AB - In order to examine the role of muscular mechanisms in chronic tension-type headache a study with needle acupuncture was performed. Needle acupuncture could be of therapeutic value because it has shown some positive effects in myofascial pain syndromes. We performed a double-blind, placebo-controlled study with 39 patients (mean age 49.0 years, SD=14.8) fulfilling the International Headache Society criteria for chronic tension-type headaches. Participants were randomly assigned to verum or placebo condition. Six weeks after end of treatment no significant differences between placebo and verum could be observed with respect to visual analogue scale and frequency of headache attacks. Nevertheless, pressure pain thresholds significantly increased for the verum group. The findings of our study support the hypothesis that peripheral mechanisms - such as increased muscle tenderness - only play a minor role in the pathogenesis of chronic tension-type headache. PMID- 11050377 TI - Introduction PMID- 11050376 TI - Cloning and functional expression of a human orthologue of rat vanilloid receptor 1. AB - Capsaicin, resiniferatoxin, protons or heat have been shown to activate an ion channel, termed the rat vanilloid receptor-1 (rVR1), originally isolated by expression cloning for a capsaicin sensitive phenotype. Here we describe the cloning of a human vanilloid receptor-1 (hVR1) cDNA containing a 2517 bp open reading frame that encodes a protein with 92% homology to the rat vanilloid receptor-1. Oocytes or mammalian cells expressing this cDNA respond to capsaicin, pH and temperature by generating inward membrane currents. Mammalian cells transfected with human VR1 respond to capsaicin with an increase in intracellular calcium. The human VR1 has a chromosomal location of 17p13 and is expressed in human dorsal root ganglia and also at low levels throughout a wide range of CNS and peripheral tissues. Together the sequence homology, similar expression profile and functional properties confirm that the cloned cDNA represents the human orthologue of rat VR1. PMID- 11050378 TI - Low-molecular-weight heparin in patients with non-ST-segment elevation acute coronary syndrome: role in the emergency department. AB - Intravenous unfractionated heparin (UFH) has been shown to be an effective therapy in reducing the risk of death or myocardial infarction in patients with unstable angina. Low molecular weight heparins demonstrate improved pharmacologic and pharmacokinetic properties relative to standard heparin, and these advantages have been translated into similar or even greater clinical efficacy in several large-scale clinical trials evaluating their use. The simple mode of administration and lack of dependency on anticoagulation monitoring make low molecular-weight heparins an extremely attractive option in the treatment of patients with acute ischemic coronary syndromes presenting without persistent ST segment elevation. PMID- 11050379 TI - Glycoprotein IIb/IIIa receptor inhibitor use in unstable angina/non-ST-segment elevation myocardial infarction: an appropriate-use model for the emergency department. AB - The introduction of new platelet glycoprotein (GP) IIb/IIIa receptor inhibitors has the potential to revolutionize the treatment of certain patients with unstable angina and non-ST-segment elevation myocardial infarction (MI). Treatment with these potent platelet inhibitors has been shown to improve patient outcomes prior to percutaneous intervention (PCI), following PCI, and even in the absence of PCI. In addition, there are data from clinical studies that support a time-dependence to the beneficial effects of platelet inhibitor therapy. Patients who are treated early-for example, in the emergency department (ED)-appear to have improved outcomes over those treated later. Platelet inhibitors have been used extensively in the cardiac catheterization laboratory and in the coronary care unit (CCU). In many institutions, however, their introduction and uptake in the ED has been slow. These agents are relatively expensive and have the potential to cause major bleeding complications. This review will examine the pathophysiology of acute coronary syndromes (ACS) as it relates to the mechanism of action of GPIIb/IIIa inhibitors. The available GPIIb/IIIa inhibitors will be discussed including their indications, contraindications, doses, and mechanisms of action. Data from clinical studies supporting the use of GPIIb/IIIa inhibitors will be illustrated in the context of their applications in the ED. Finally, an algorithm for GPIIb/IIIa inhibitors in the ED will be presented. PMID- 11050380 TI - ER TIMI-19: testing the reality of prehospital thrombolysis. PMID- 11050381 TI - Facilitated percutaneous coronary intervention for acute myocardial infarction. AB - Facilitated percutaneous coronary intervention is a treatment strategy for acute myocardial infarction in which patients are given medications in the emergency department that open or partially open infarct-related coronary arteries. The patients are then taken to the cardiac catheterization laboratory for early angiography and angioplasty or placement of a coronary artery stent. Preliminary evidence suggests that this treatment strategy may offer outcomes similar to or better than primary angioplasty and superior to solitary fibrinolytic therapy. In addition, the treatment can be started even in hospitals that do not have primary intervention capability. Currently, large-scale clinical trials are assessing the impact of the facilitated percutaneous coronary intervention treatment strategy on the clinical outcomes of patients with acute myocardial infarction. PMID- 11050382 TI - Combination therapy: management of acute myocardial infarction in the new millennium. PMID- 11050383 TI - Brain-derived neurotrophic factor is an anterograde survival factor in the rat visual system. AB - BACKGROUND: The neurotrophins, which include nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), NT-4/5 and NT-6, are a family of proteins that play fundamental roles in the differentiation, survival and maintenance of peripheral and central neurons. Much research has focused on the role of neurotrophins as target-derived, retrogradely transported trophic molecules. Although there is recent evidence that BDNF and NT-3 can be transported in an anterograde direction along peripheral and central axons, there is as yet no conclusive evidence that these anterograde factors have direct post synaptic actions. RESULTS: We report that BDNF travels in an anterograde direction along the optic nerve. The anterogradely transported BDNF had rapid effects on retinal target neurons in the superior colliculus and lateral geniculate nucleus of the brain. When endogenous BDNF within the developing superior colliculus was neutralised, the rate of programmed neuronal death increased. Conversely, provision of an afferent supply of BDNF prevented the degeneration of geniculate neurons after removal of their cortical target. CONCLUSIONS: BDNF released from retinal ganglion cells acts as a survival factor for post-synaptic neurons in retinal target fields. PMID- 11050384 TI - The aurora-related kinase AIR-2 recruits ZEN-4/CeMKLP1 to the mitotic spindle at metaphase and is required for cytokinesis. AB - BACKGROUND: The Aurora/Ipl1p-related kinase AIR-2 is required for mitotic chromosome segregation and cytokinesis in early Caenorhabditis elegans embryos. Previous studies have relied on non-conditional mutations or RNA-mediated interference (RNAi) to inactivate AIR-2. It has therefore not been possible to determine whether AIR-2 functions directly in cytokinesis or if the cleavage defect results indirectly from the failure to segregate DNA. One intriguing hypothesis is that AIR-2 acts to localize the mitotic kinesin-like protein ZEN-4 (also known as CeMKLP1), which later functions in cytokinesis. RESULTS: Using conditional alleles, we established that AIR-2 is required at metaphase or early anaphase for normal segregation of chromosomes, localization of ZEN-4, and cytokinesis. ZEN-4 is first required late in cytokinesis, and also functions to maintain cell separation through much of the subsequent interphase. DNA segregation defects alone were not sufficient to disrupt cytokinesis in other mutants, suggesting that AIR-2 acts specifically during cytokinesis through ZEN 4. AIR-2 and ZEN-4 shared similar genetic interactions with the formin homology (FH) protein CYK-1, suggesting that AIR-2 and ZEN-4 function in a single pathway, in parallel to a contractile ring pathway that includes CYK-1. Using in vitro co immunoprecipitation experiments, we found that AIR-2 and ZEN-4 interact directly. CONCLUSIONS: AIR-2 has two functions during mitosis: one in chromosome segregation, and a second, independent function in cytokinesis through ZEN-4. AIR 2 and ZEN-4 may act in parallel to a second pathway that includes CYK-1. PMID- 11050385 TI - Incenp and an aurora-like kinase form a complex essential for chromosome segregation and efficient completion of cytokinesis. AB - BACKGROUND: In animal cells, cytokinesis begins shortly after the sister chromatids move to the spindle poles. The inner centromere protein (Incenp)has been implicated in both chromosome segregation and cytokinesis, but it is not known exactly how it mediates these two distinct processes. RESULTS: We identified two Caenorhabditis elegans proteins, ICP-1 and ICP-2, with significant homology in their carboxyl termini to the corresponding region of vertebrate Incenp. Embryos depleted of ICP-1 by RNA-mediated interference had defects in both chromosome segregation and cytokinesis. Depletion of the Aurora-like kinase AIR-2 resulted in a similar phenotype. The carboxy-terminal region of Incenp is also homologous to that in Sli15p, a budding yeast protein that functions with the yeast Aurora kinase Ipl1p. ICP-1 bound C. elegans AIR-2 in vitro, and the corresponding mammalian orthologs Incenp and AIRK2 could be co-immunoprecipitated from cell extracts. A significant fraction of embryos depleted of ICP-1 and AIR-2 completed one cell division over the course of several cell cycles. ICP-1 promoted the stable localization of ZEN-4 (also known as CeMKLP1), a kinesin-like protein required for central spindle assembly. CONCLUSIONS: ICP-1 and AIR-2 are part of a complex that is essential for chromosome segregation and for efficient completion of cytokinesis. We propose that this complex acts by promoting dissolution of sister chromatid cohesion and the assembly of the central spindle. PMID- 11050386 TI - Slk19p is necessary to prevent separation of sister chromatids in meiosis I. AB - BACKGROUND: A fundamental difference between meiotic and mitotic chromosome segregation is that in meiosis I, sister chromatids remain joined, moving as a unit to one pole of the spindle rather than separating as they do in mitosis. It has long been known that the sustained linkage of sister chromatids through meiotic anaphase I is accomplished by association of the chromatids at the centromere region. The localization of the cohesin Rec8p to the centromeres is essential for maintenance of sister chromatid cohesion through meiosis I, but the molecular basis for the regulation of Rec8p and sister kinetochores in meiosis remains a mystery. RESULTS: We show that the SLK19 gene product from Saccharomyces cerevisiae is essential for proper chromosome segregation during meiosis I. When slk19 mutants were induced to sporulate they completed events characteristic of meiotic prophase I, but at the first meiotic division they segregated their sister chromatids to opposite poles at high frequencies. The vast majority of these cells did not perform a second meiotic division and proceeded to form dyads (asci containing two spores). Slk19p was found to localize to centromere regions of chromosomes during meiotic prophase where it remained until anaphase I. In the absence of Slk19p, Rec8p was not maintained at the centromere region through anaphase I as it is in wild-type cells. Finally, we demonstrate that Slk19p appears to function downstream of the meiosis-specific protein Spo13p in control of sister chromatid behavior during meiosis I. CONCLUSIONS: Our results suggest that Slk19p is essential at the centromere of meiotic chromosomes to prevent the premature separation of sister chromatids at meiosis I. PMID- 11050387 TI - Evidence that processed small dsRNAs may mediate sequence-specific mRNA degradation during RNAi in Drosophila embryos. AB - BACKGROUND: RNA interference (RNAi) is a phenomenon in which introduced double stranded RNAs (dsRNAs) silence gene expression through specific degradation of their cognate mRNAs. Recent analyses in vitro suggest that dsRNAs may be copied, or converted, into 21-23 nucleotide (nt) guide RNAs that direct the nucleases responsible for RNAi to their homologous mRNA targets. Such small RNAs are also associated with gene silencing in plants. RESULTS: We developed a quantitative single-embryo assay to examine the mechanism of RNAi in vivo. We found that dsRNA rapidly induced mRNA degradation. A fraction of dsRNAs were converted into 21-23 nt RNAs, and their time of appearance and persistence correlated precisely with inhibition of expression. The strength of RNAi increased disproportionately with increasing dsRNA length, but an 80bp dsRNA was capable of effective gene silencing. RNAi was saturated at low dsRNA concentration and inhibited by excess unrelated dsRNA. The antisense strand of the dsRNA determined target specificity, and excess complementary sense or antisense single-stranded RNAs (ssRNAs) competed with the RNAi reaction. CONCLUSIONS: Processed dsRNAs can act directly to mediate RNAi, with the antisense strand determining mRNA target specificity. The involvement of 21-23 nt RNAs is supported by the kinetics of the processing reaction and the observed size dependence. RNAi depends on a limiting factor, possibly the nuclease that generates the 21-23 mer species. The active moiety appears to contain both sense and antisense RNA strands. PMID- 11050388 TI - Expression of the pro-apoptotic Bcl-2 family member Bim is regulated by the forkhead transcription factor FKHR-L1. AB - Cell death is regulated mainly through an evolutionarily conserved form of cell suicide termed apoptosis [1]. Deregulation of apoptosis has been associated with cancer, autoimmune diseases and degenerative disorders. Many cells, particularly those of the hematopoietic system, have a default program of cell death and survival that is dependent on the constant supply of survival signals. The Bcl-2 family, which has both pro- and anti-apoptotic members, plays a critical role in regulating cell survival [2]. One family member, the Bcl-2 interacting mediator of cell death (Bim), contains only a protein-interaction motif known as the BH3 domain, allowing it to bind pro-survival Bcl-2 molecules, neutralizing their function [3]. Disruption of the bim gene results in resistance to apoptosis following cytokine withdrawal in leukocytes, indicating that regulation of the pro-apoptotic activity of Bim is critical for maintenance of the default apoptotic program [4]. Here, we report that withdrawal of cytokine results in upregulation of Bim expression concomitant with induction of the apoptotic program in lymphocytes. Activation of the forkhead transcription factor FKHR-L1, previously implicated in regulation of apoptosis in T lymphocytes [5], was sufficient to induce Bim expression. We propose a mechanism by which cytokines promote lymphocyte survival by inhibition of FKHR-L1, preventing Bim expression. PMID- 11050389 TI - A 'non-canonical' DNA-binding element mediates the response of the Fas-ligand promoter to c-Myc. AB - Cell number is regulated by maintaining a balance between cell proliferation and cell death through apoptosis. Key regulators of this balance include the oncogene product c-Myc, which promotes either entry into the cell cycle or apoptosis [1]. Although the mechanism of c-Myc-induced apoptosis remains unclear, it is susceptible to regulation by survival factors [2,3] and can proceed through the interaction of Fas ligand (FasL) with its receptor, Fas [4]. Activated T lymphocytes are eliminated by an apoptotic process known as activation-induced cell death (AICD), which requires the transcriptional induction of FasL expression [5-7] and sustained levels of c-Myc [8]. The FasL promoter can be driven by c-Myc overexpression, and functional inhibitors of Myc and its binding partner, Max, inhibit the transcriptional activity of the FasL promoter [9,10]. We identified a non-canonical binding site (ATTCTCT) for c-Myc-Max heterodimers in the FasL promoter, which, when mutated, abolished activity in response to c Myc. Exchange of the canonical c-Myc responsive elements (CACGTG) in the ornithine decarboxylase (ODC) promoter [11] with the non-canonical sequence in the FasL promoter generated an ODC-FasL promoter that was significantly more responsive to c-Myc than the wild-type ODC promoter. Our findings identify a precise physiological role for c-Myc in the induction of apoptosis as a transcriptional regulator of the FasL gene. PMID- 11050390 TI - Refixation frequency and memory mechanisms in visual search. AB - Visual search-looking for a target object in the presence of a number of distractor items-is an everyday activity for humans (for example, finding the car in a busy car park) and animals (for example, foraging for food). Our understanding of visual search has been enriched by an interdisciplinary effort using a wide range of research techniques including behavioural studies in humans [1], single-cell electrophysiology [2], transcranial magnetic stimulation [3], event-related potentials [4] and studies of patients with focal brain injury [5]. A central question is what kind of information controls the search process. Visual search is typically accompanied by a series of eye movements, and investigating the nature and location of fixations helps to identify the kind of information that might control the search process. It has already been demonstrated that objects are fixated if they are visually similar to the target [6]. Also, if an item has been fixated, it is less likely to be returned to on the subsequent saccade. This automatic process is referred to as inhibition of return (IOR [7,8]). Here, we investigated the role of memory for which items had been fixated previously. We found that, during search, subjects often refixated items that had been previously fixated. Although there were fewer return saccades than would be expected by chance, the number of refixations indicated limited functional memory, indeed the memory effects that were present may primarily be a result of IOR. PMID- 11050391 TI - Disruption of mouse polymerase zeta (Rev3) leads to embryonic lethality and impairs blastocyst development in vitro. AB - Multiple DNA polymerases exist in eukaryotes. Polymerases alpha, delta and epsilon are mainly responsible for chromosomal DNA replication in the nucleus and are required for proliferation. In contrast, the repair polymerases beta and eta are not essential for cellular proliferation in yeast or mice, but a lack of either polymerase can lead, respectively, to defects in base excision repair or the ability to replicate past lesions induced by ultraviolet (UV) radiation [1 3]. Here, we have focused on polymerase zeta. This was first described as a non essential product of the yeast REV3/REV7 genes involved in UV-induced mutagenesis, and was later implicated in trans-lesion synthesis [4,5]. Unlike in yeast, the mouse homologue (mRev3) was found to be essential for life. Homozygous mutant mice died in utero. Mutant embryos were considerably reduced in size at day 10.5 of development and usually aborted around day 12.5. It is likely that this block reflects a need for mRev3 in proliferative clonal expansion (rather than in the production of a particular cell type) as mutant blastocysts showed greatly diminished expansion of the inner cell mass in culture. Thus, mRev3 could be required to repair a form of externally induced DNA damage that otherwise accumulates during clonal expansion or, consistent with the high homology shared between its Rev7 partner and the mitotic checkpoint gene product Mad2 [6], mRev3 might play a role in cell proliferation and genomic stability even in the absence of environmentally induced damage. PMID- 11050392 TI - Disruption of the developmentally regulated Rev3l gene causes embryonic lethality. AB - The REV3 gene encodes the catalytic subunit of DNA polymerase (pol) zeta, which can replicate past certain types of DNA lesions [1]. Saccharomyces cerevisiae rev3 mutants are viable and have lower rates of spontaneous and DNA-damage induced mutagenesis [2]. Reduction in the level of Rev31, the presumed catalytic subunit of mammalian pol zeta, decreased damage-induced mutagenesis in human cell lines [3]. To study the function of mammalian Rev31, we inactivated the gene in mice. Two exons containing conserved DNA polymerase motifs were replaced by a cassette encoding G418 resistance and beta-galactosidase, under the control of the Rev3l promoter. Surprisingly, disruption of Rev3l caused mid-gestation embryonic lethality, with the frequency of Rev3l(-/-) embryos declining markedly between 9.5 and 12.5 days post coitum (dpc). Rev3l(-/-) embryos were smaller than their heterozygous littermates and showed retarded development. Tissues in many areas were disorganised, with significantly reduced cell density. Rev3l expression, traced by beta-galactosidase staining, was first detected during early somitogenesis and gradually expanded to other tissues of mesodermal origin, including extraembryonic membranes. Embryonic death coincided with the period of more widely distributed Rev3l expression. The data demonstrate an essential function for murine Rev31 and suggest that bypass of specific types of DNAlesions by pol zeta is essential for cell viability during embryonic development in mammals. PMID- 11050393 TI - Disruption of the Rev3l-encoded catalytic subunit of polymerase zeta in mice results in early embryonic lethality. AB - Polymerase zeta (Pol zeta) is an error-prone DNA polymerase [1], which in yeast is involved in trans-lesion synthesis (TLS) and is responsible for most of the ultraviolet (UV) radiation-induced and spontaneous mutagenesis [2-4]. Pol zeta consists of three subunits: REV1, a deoxycytidyl-transferase [5]; REV7, of unclear function [6]; and REV3, the catalytic subunit. REV3 alone is sufficient to carry out TLS, but association with REV1 and REV7 enhances its activity [5, 7]. Experiments using human cells treated with UV radiation indicate also that mammalian Pol zeta is involved in TLS [7]. The peculiar mutagenic activity of Pol zeta [4,7,8] suggests a possible role in somatic hypermutation of immunoglobulin (Ig) genes [9]. Here, we report that, unlike in yeast where the REV3 gene is not essential for life [4], disruption of the mouse homologue (Rev3l) resulted in early embryonic lethality. In Rev3l(-/-) embryos, no haematopoietic cells other than erythrocytes could be identified in the yolk sac. Rev3l(-/-) haematopoietic precursors were unable to expand in vitro and no haematopoietic cells could be derived from the intraembryonic haematogenic compartment (splanchnopleura). Fibroblasts could not be derived from the Rev3l(-/-) embryos, and Rev3l(-/-) embryonic stem (ES) cells could not be obtained. This is the first evidence that an enzyme involved in TLS is critical for mammalian development. PMID- 11050394 TI - A permissive function of phosphoinositide 3-kinase in Ras activation mediated by inhibition of GTPase-activating proteins. AB - The activation status of the guanosine triphosphate (GTP)-binding protein Ras is dictated by the relative intensities of two opposing reactions: the formation of active Ras-GTP complexes, promoted by guanine-nucleotide exchange factors (GEFs), and their conversion to inactive Ras-GDP as a result of the deactivating action of GTPase-activating proteins (GAPs). The relevance of phosphoinositide 3-kinase (PI 3-kinase) to these processes is still unclear. We have investigated the regulation of Ras activation by PI 3-kinase in the myelomonocytic U937 cell line. These cells exhibited basal levels of Ras-GTP, which were suppressed by two PI 3 kinase inhibitors and a dominant-negative PI 3-kinase. In addition, PI 3-kinase inhibition aborted Ras activation by all stimuli tested, including foetal calf serum (FCS) and phorbol 12-myristate 13-acetate (TPA). Significantly, TPA does not activate PI 3-kinase in U937 cells, indicating that PI 3-kinase has a permissive rather than an intermediary role in Ras activation. Investigation of the mechanism of PI 3-kinase action revealed that inhibition of PI 3-kinase does not affect nucleotide exchange on Ras but abrogates Ras-GTP accumulation through an increase in GAP activity. These findings establish blockage of GAP action as the mechanism underlying a permissive function of PI 3-kinase in Ras activation. PMID- 11050395 TI - Roles for polarity and nuclear determinants in specifying daughter cell fates after an asymmetric cell division in the maize leaf. AB - Asymmetric cell divisions occur repeatedly during plant development, but the mechanisms by which daughter cells are directed to adopt different fates are not well understood [1,2]. Previous studies have demonstrated roles for positional information in specification of daughter cell fates following asymmetric divisions in the embryo [3] and root [4]. Unequally inherited cytoplasmic determinants have also been proposed to specify daughter cell fates after some asymmetric cell divisions in plants [1,2,5], but direct evidence is lacking. Here we investigate the requirements for specification of stomatal subsidiary cell fate in the maize leaf by analyzing four mutants disrupting the asymmetric divisions of subsidiary mother cells (SMCs). We show that subsidiary cell fate does not depend on proper localization of the new cell wall during the SMC division, and is not specified by positional information acting on daughter cells after completion of the division. Instead, our data suggest that specification of subsidiary cell fate depends on polarization of SMCs and on inheritance of the appropriate daughter nucleus. We thus provide evidence of a role for unequal inheritance of an intracellular determinant in specification of cell fate after an asymmetric plant cell division. PMID- 11050396 TI - Identification of CISK, a new member of the SGK kinase family that promotes IL-3 dependent survival. AB - The signaling pathways for cell survival are much less well understood than those for apoptosis [1]. Many mammalian cell-survival factors have been identified, either biochemically or from genetic studies in other organisms. Effective genetic methods that allow systematic study of anti-apoptosis genes in mammalian cells remain to be established, however. To achieve this goal, we used a new genetic screening method using enhanced retroviral mutagen (ERM) vectors to identify factors that mediate IL-3-dependent survival of hematopoietic cells. Both known and novel mediators of cell survival were identified, including Bcl xL, phosphatidylinositide 3-kinase (PI 3-kinase), Akt and cytokine-independent survival kinase (CISK). CISK is a PX-domain-containing serine/threonine kinase homologous to serum- and glucocorticoid-regulated protein kinase (SGK). We showed that CISK acts downstream of the PI 3-kinase cascade in vivo and may function in parallel to Akt by phosphorylating Bad and the transcription factor FKHRL1. The distinct subcellular localization of CISK, however, suggests that it acts in different signaling cascades from Akt. Our results demonstrate the power of ERM to identify key genes involved in cell-survival signaling. Furthermore, CISK is the first SGK family member that has been shown to promote survival, pointing to the possibility that other SGK family proteins may also function in survival pathways. PMID- 11050398 TI - Brain story PMID- 11050397 TI - My word. Will biotech venture thrive in the new climate in Japan? PMID- 11050399 TI - Mariano barbacid: bringing cancer biology back home PMID- 11050400 TI - Quick guide. Matrix metalloproteinases. PMID- 11050401 TI - Providencia may help find a function for a novel, widespread protein family. PMID- 11050402 TI - Actin cytoskeleton: putting a CAP on actin polymerization. AB - Two recent studies have identified a Drosophila homolog of cyclase-associated protein (CAP) as a developmentally important negative regulator of actin polymerization that may also directly mediate signal transduction. PMID- 11050403 TI - Chromosome cohesion: a polymerase for chromosome bridges. AB - How do cells ensure that sister chromatids produced during DNA replication stay connected with each other until their separation in anaphase? New insight is provided by the discovery of DNA polymerase kappa, which has been found to be required for building the connections between sister chromatids. PMID- 11050404 TI - Stereoscopic vision: what's the first step? AB - Neurons in primary visual cortex respond to binocular disparity, the raw material of stereoscopic depth perception. Although these neurons are probably essential to depth perception, a recent study has shown that they are unable to compute depth itself. PMID- 11050405 TI - Cooperativity: action at a distance in a classic system. AB - A new high resolution crystal structure of the phage lambda repressor reveals the basis for repressor dimer formation and, together with biochemical data, provides insights into the mechanism of repressor tetramer formation, a process essential to the cooperative binding and gene regulatory activities of this protein. PMID- 11050406 TI - Gene silencing: two faces of SIR2. AB - There are still many mysteries surrounding how silenced regions of the eukaryotic genome are created and maintained. But recent discoveries about the most evolutionarily conserved silencing protein, Sir2p, have provided new mechanistic insights into these processes. PMID- 11050407 TI - Neuroglial networks: neurons and glia talk to each other. AB - Recent results show that certain neurons and glia are connected by gap junctions, and that neurons can regulate gap-junctional communication by glia. The findings are inspiring a major reexamination of the role of glia in the regulation of neuronal integration in the central nervous system. PMID- 11050408 TI - Translation initiation: insect virus RNAs rewrite the rule book. AB - Picorna-like insect virus RNAs direct an unorthodox form of translation initiation at a non-AUG-related codon, without involvement of initiator tRNA. This seems to involve a special type of mRNA pseudoknot structure which allows bypassing of the usual P-site-dependent mechanism. PMID- 11050409 TI - Animal navigation: birds as geometers? AB - New experiments on a bird species able to remember the sites of thousands of cached seeds have revealed how a site can be specified by combining distance information from several landmarks. PMID- 11050410 TI - Human evolution: how recent were the Y chromosome ancestors? AB - Recent findings of low sequence variability of Y chromosome genes has led to suggestions that the most recent ancestor of human Y chromosomes existed around 50,000 years ago and human population size expanded about 28,000 years ago. But what level of confidence can we have in these estimates? PMID- 11050412 TI - Gone with the wnts PMID- 11050411 TI - Jamming the endosomal system: lipid rafts and lysosomal storage diseases. AB - Some lysosomal storage diseases result from the accumulation of lipids in degradative compartments of the endocytic pathway. Particularly striking is the example of the Niemann-Pick (NP) syndrome. NP syndromes types A and B are characterized by the accumulation of sphingomyelin, whereas cholesterol typically accumulates in NP type C. These two different lipids, sphingomyelin and cholesterol, are normal constituents of specific lipid microdomains called rafts. Because accumulation of raft lipids is observed not only in NP diseases but also in many other lipidoses, we forward the hypothesis that lysosomal storage diseases can be caused by the accumulation of lipid rafts in late endosomes/lysosomes. PMID- 11050414 TI - It takes (more than) two to tango PMID- 11050413 TI - Placing Ena/VASP proteins onto the circle of actin dynamics PMID- 11050416 TI - Liberating sleeping beauties by cutting the hook PMID- 11050415 TI - Visions of IFT PMID- 11050417 TI - The unpredictability of PI 3-kinase PMID- 11050418 TI - Leukocytes navigate by compass: roles of PI3Kgamma and its lipid products. AB - Morphologic polarity is necessary for the motility of mammalian cells. In leukocytes responding to a chemoattractant, this polarity is regulated by activities of small Rho guanosine triphosphatases (Rho GTPases) and the phosphoinositide 3-kinases (PI3Ks). Moreover, in neutrophils, lipid products of PI3Ks appear to regulate activation of Rho GTPases, are required for cell motility and accumulate asymmetrically to the plasma membrane at the leading edge of polarized cells. By spatially regulating Rho GTPases and organizing the leading edge of the cell, PI3Ks and their lipid products could play pivotal roles not only in establishing leukocyte polarity but also as compass molecules that tell the cell where to crawl. PMID- 11050419 TI - Making memories stick: cell-adhesion molecules in synaptic plasticity. AB - Synapses are adhesive junctions highly specialized for interneuronal signalling in the central nervous system. The strength of the synaptic signal can be modified (synaptic plasticity), a key feature of the cellular changes thought to underlie learning and memory. Cell-adhesion molecules are important constituents of synapses, with well-recognized roles in building and maintaining synaptic structure during brain development. However, growing evidence indicates that cell adhesion molecules also play important and diverse roles in regulating synaptic plasticity and learning and memory. This review focuses on recent advances in understanding the molecular mechanisms through which adhesion molecules might regulate synaptic plasticity. PMID- 11050420 TI - Proteins on the move: dynamic protein localization in prokaryotes. AB - Despite their small size and lack of obvious intracellular structures, bacteria have a complex and dynamic intracellular organization. Recent work has shown that many proteins, and even regions of the chromosome, are localized to specific subcellular regions that can change over time, sometimes extraordinarily fast. Protein function can depend on cellular position, so the analysis of the intracellular location of a protein can be crucial for understanding its activity. Because regulatory proteins are among those that reside at specific cellular sites, it is now necessary to consider three-dimensional organization when describing the genetic networks that control bacterial cells. PMID- 11050421 TI - The biology and analysis of single disseminated tumour cells. AB - Despite an increasing molecular-genetic understanding of the development of malignant epithelial neoplasias, the frontline therapy for patients with carcinomas is still surgery. Systemic adjuvant treatments such as chemotherapy or immunotherapy have had limited success perhaps because they are based on analysis of the primary tumour or on cell lines derived from metastasis. However, the characteristics of systemically disseminated tumour cells can be very different from that of the primary tumour or end-stage metastasis. Consequently, there is a need to study the evolution and nature of systemic cancer directly in order to identify new target structures for therapy present on the potential precursors of metastasis--the disseminated tumour cells. PMID- 11050422 TI - Hans Ris--from chromatin fibres, through nuclear tracks, to nuclear pores. PMID- 11050423 TI - Applying for grant funds: there's help around the corner. AB - Scientists should know where to get help when they write grant proposals. Books, videos, consultants and colleagues can all be useful. The World Wide Web is an excellent resource for finding competitors, collaborators and a wealth of other useful information such as funding agencies, available grant money, pertinent articles and form pages for grant applications. Software is available to help you make an outline of your proposal, prepare the required figures and tables and build a database of references from which to create a bibliography. This article will point you in the right direction for a plethora of information and resources. PMID- 11050424 TI - Novel homologs of gp91phox. PMID- 11050425 TI - TROSY and CRINEPT: NMR with large molecular and supramolecular structures in solution. AB - TROSY and CRINEPT are new techniques for solution NMR studies of molecular and supramolecular structures. They allow the collection of high-resolution spectra of structures with molecular weights >100 kDa, significantly extending the range of macromolecular systems that can be studied by NMR in solution. TROSY has already been used to map protein-protein interfaces, to conduct structural studies on membrane proteins and to study nucleic acid conformations in multimolecular assemblies. These techniques will help us to investigate the conformational states of individual macromolecular components and will support de novo protein structure determination in large supramolecular structures. PMID- 11050426 TI - Erratum PMID- 11050427 TI - BRCA1: exploring the links to transcription. AB - Progress on determining the function of the breast and ovarian cancer susceptibility gene BRCA1 suggests it might be involved in two fundamental cellular processes: DNA repair and transcriptional regulation. Recent developments indicate that BRCA1 is a multifunctional protein, and disruption of its transcriptional activity could be crucial for tumor development. PMID- 11050428 TI - Gene context conservation of a higher order than operons. AB - Operons, co-transcribed and co-regulated contiguous sets of genes, are poorly conserved over short periods of evolutionary time. The gene order, gene content and regulatory mechanisms of operons can be very different, even in closely related species. Here, we present several lines of evidence which suggest that, although an operon and its individual genes and regulatory structures are rearranged when comparing the genomes of different species, this rearrangement is a conservative process. Genomic rearrangements invariably maintain individual genes in very specific functional and regulatory contexts. We call this conserved context an uber-operon. PMID- 11050429 TI - Domains in gene silencing and cell differentiation proteins: the novel PAZ domain and redefinition of the Piwi domain. PMID- 11050431 TI - Condensing the RNA world PMID- 11050430 TI - GRAM, a novel domain in glucosyltransferases, myotubularins and other putative membrane-associated proteins. PMID- 11050432 TI - Fringe gives a saccharine to notch PMID- 11050433 TI - Joining tethers and SNAREs PMID- 11050434 TI - Cytohesins and centaurins: mediators of PI 3-kinase-regulated Arf signaling. AB - Receptor-activated phosphoinositide (PI) 3-kinases produce PtdIns(3, 4,5)P(3) and its metabolite PtdIns(3,4)P(2) that function as second messengers in membrane recruitment and activation of target proteins. The cytohesin and centaurin protein families are potential targets for PtdIns(3,4,5)P(3) that also regulate and interact with Arf GTPases. Consequently, these families are poised to transduce PI 3-kinase activation into coordinated control of Arf-dependent pathways. Proposed downstream events in PI 3-kinase-regulated Arf cascades include modulation of vesicular trafficking and the actin cytoskeleton. PMID- 11050435 TI - STAT proteins and transcriptional responses to extracellular signals. AB - Signal transducer and activator of transcription (STAT) transcription factors are implicated in programming gene expression in biological events as diverse as embryonic development, programmed cell death, organogenesis, innate immunity, adaptive immunity and cell growth regulation in organisms ranging from slime molds to insects to man. Rapid progress has unearthed much about the activation of STATs by Janus kinases (JAKs) and other tyrosine kinases and their ability to interface with other signaling systems. Once inside the nucleus, the STATs bind to promoters and join other transcriptional activators in the regulation of gene expression. PMID- 11050436 TI - Mitochondria, oxygen free radicals, disease and ageing. AB - Superoxide is generated by the mitochondrial respiratory chain. The transformation of this superoxide into hydrogen peroxide and, under certain conditions, then into hydroxyl radicals is important in diseases where respiratory chain function is abnormal or where superoxide dismutase function is altered, as in amyotrophic lateral sclerosis. In addition, these reactive oxygen species can influence the ageing process through mechanisms involving mutagenesis of mtDNA or increased rates of shortening of telomeric DNA. PMID- 11050438 TI - Web alert. Antimicrobials genomics. PMID- 11050437 TI - When protein folding is simplified to protein coiling: the continuum of solenoid protein structures. AB - Solenoid proteins contain repeating structural units that form a continuous superhelix. This category of proteins conveys the least complicated relationship between a sequence and the corresponding three-dimensional structure. Although solenoid proteins are divided into different classes according to commonly used classification schemes, they share many structural and functional properties. PMID- 11050439 TI - The genomics of microbial diversity. Editorial overview PMID- 11050440 TI - Reverse vaccinology. AB - Biochemical, serological and microbiological methods have been used to dissect pathogens and identify the components useful for vaccine development. Although successful in many cases, this approach is time-consuming and fails when the pathogens cannot be cultivated in vitro, or when the most abundant antigens are variable in sequence. Now genomic approaches allow prediction of all antigens, independent of their abundance and immunogenicity during infection, without the need to grow the pathogen in vitro. This allows vaccine development using non conventional antigens and exploiting non-conventional arms of the immune system. Many vaccines impossible to develop so far will become a reality. Since the process of vaccine discovery starts in silico using the genetic information rather than the pathogen itself, this novel process can be named reverse vaccinology. PMID- 11050441 TI - Signature-tagged mutagenesis in the identification of virulence genes in pathogens. AB - Signature-tagged mutagenesis is a functional genomics technique that identifies microbial genes required for infection within an animal host, or within host cells. The application of this technique to a range of microbial pathogens has resulted in the identification of novel virulence determinants in each screen performed to date, so that cumulatively several hundred genes have been ascribed a role in virulence. PMID- 11050442 TI - A genomic approach to the understanding of Xylella fastidiosa pathogenicity. AB - Xylella fastidiosa is a fastidious, xylem-limited bacterium that causes several economically important plant diseases, including citrus variegated chlorosis (CVC). X. fastidiosa is the first plant pathogen to have its genome completely sequenced. In addition, it is probably the least previously studied of any organism for which the complete genome sequence is available. Several pathogenicity-related genes have been identified in the X. fastidiosa genome by similarity with other bacterial genes involved in pathogenesis in plants, as well as in animals. The X. fastidiosa genome encodes different classes of proteins directly or indirectly involved in cell-cell interactions, degradation of plant cell walls, iron homeostasis, anti-oxidant responses, synthesis of toxins, and regulation of pathogenicity. Neither genes encoding members of the type III protein secretion system nor avirulence-like genes have been identified in X. fastidiosa. PMID- 11050443 TI - Towards the minimal eukaryotic parasitic genome. AB - Microsporidia are well-known to infect immunocompromised patients and are also responsible for clinical syndromes in immunocompetent individuals. In recent years, evidence has been obtained in support of a very close relationship between Microsporidia and Fungi. In some species, the compaction of the genome and genes is remarkable. Thus, a systematic sequencing project has been initiated for the 2.9 Mbp genome of Encephalitozoon cuniculi, which will be useful for future comparative genomic studies. PMID- 11050444 TI - Molecular genetics in Aspergillus fumigatus. AB - Manipulation of the genome of the human pathogen Aspergillus fumigatus is not well developed. Approaches and data from related model organisms are being used to develop molecular genetic systems in A. fumigatus; for example, the molecular typing of strains during infection. A genome-sequencing programme has begun and will form the basis for future development. PMID- 11050445 TI - Assessing evolutionary relationships among microbes from whole-genome analysis. AB - The determination and analysis of complete genome sequences have recently enabled many major advances to be made in the area of microbial evolutionary biology. These include the determination of the first genome of a Crenarchaeota, the suggestion that horizontal gene transfer may be the rule rather than the exception, and revelations about how genomes evolve on short timescales. PMID- 11050446 TI - Origins of hydrogenosomes and mitochondria. AB - Complete genome sequences for many oxygen-respiring mitochondria, as well as for some bacteria, leave no doubt that mitochondria are descendants of alpha proteobacteria, a finding for which the endosymbiont hypothesis can easily account. Yet a wealth of data indicate that mitochondria and hydrogenosomes - the ATP-producing organelles of many anaerobic protists - share a common ancestry, a finding that traditional formulations of the endosymbiont hypothesis less readily accommodates. Available evidence suggests that a more in-depth understanding of the origins of eukaryotes and their organelles will hinge upon data from the genomes of protists that synthesize ATP without the need for oxygen. PMID- 11050447 TI - Antimicrobials: better use, better drugs, or both? PMID- 11050448 TI - Carbapenemases: a problem in waiting? AB - Carbapenems are stable to most prevalent beta-lactamases, and chromosomal carbapenemases are restricted to Stenotrophomonas maltophilia, to a few Bacteroides fragilis, and to rare pathogens. Nevertheless, an acquired metallo beta-lactamase called IMP-1 is beginning to emerge in Pseudomonas aeruginosa and Enterobacteriaceae isolates in Japan, and has also been found in isolates from Singapore. Furthermore, IMP-producing Acinetobacter spp. have been identified in Italy and Hong Kong. Recently a second group of acquired metallo-carbapenemases, the VIM types, has been recorded from P. aeruginosa isolates in five Eurasian countries. Weak carbapenemases belonging to molecular class D are emerging in A. baumannii world-wide, with two sub-groups apparent. A few acquired carbapenemases belonging to molecular class A also have been reported. Finally it has also been shown that enzymes with feeble carbapenemase activity (e.g. AmpC types and some SHV enzymes) may confer resistance in exceptionally impermeable strains; counterwise, even potent carbapenemases, such as IMP-1, may only give a small reduction in susceptibility in Enterobacteriaceae that lack permeability lesions. Is the emergence of carbapenemase a problem waiting to happen? PMID- 11050449 TI - Antimicrobial use and bacterial resistance. AB - The current epidemic of bacterial resistance is attributed, in part, to the overuse of antibiotics. Recent studies have documented increases in resistance with over-use of particular antibiotics and improvements in susceptibility when antibiotic use is controlled. The most effective means of improving use of antibiotics is unknown. Comprehensive management programs directed by multi disciplinary teams, computer-assisted decision-making, and antibiotic cycling have been beneficial in controlling antibiotic use, decreasing costs without impacting patient outcomes, and possibly decreasing resistance. PMID- 11050450 TI - Pneumococcal conjugate vaccines - implications for community antibiotic prescribing. AB - Infections caused by Streptococcus pneumoniae are a major cause of morbidity and mortality in the pediatric population worldwide. Development of increasing resistance to multiple classes of antibiotics is making treatment of infections caused by this organism much more difficult. In order to prevent disease, a 23 valent pneumococal polysaccharide vaccine is available. However, this vaccine is poorly immunogenic in infants and young children. The development and licensing of pneumococcal conjugate vaccines that are safe and effective in the pediatric population is an important step in our ability to decrease the prevalence of pneumococcal disease seen. PMID- 11050451 TI - Antiretroviral therapy of HIV-1 infection: established treatment strategies and new therapeutic options. AB - Recently, studies have shown that non-nucleoside reverse transcriptase inhibitors, such as efavirenz or nevirapine, in combination with two nucleoside analogues have an antiretroviral potency comparable to protease inhibitor containing regimens. Triple combination therapy that includes a non-nucleoside reverse transcriptase inhibitor can therefore be regarded as an effective alternative first-line treatment of HIV-1 infection. PMID- 11050452 TI - Pharmacodynamics in the study of drug resistance and establishing in vitro susceptibility breakpoints: ready for prime time. AB - Considerable advancements have been made in providing informative, relevant interpretations of the results of antimicrobial susceptibility tests to clinicians, clinical microbiologists, epidemiologists, and researchers. At the same time, the science of pharmacokinetics has flourished, and the importance of drug exposure in vivo on outcome is now recognized by researchers and clinicians alike. More recently, pharmacokinetic and quantitative measures of antimicrobial susceptibility have been integrated using pharmacokinetic-pharmacodynamic (PK-PD) models to forecast clinical and microbiological outcomes. Stochastic methods utilizing patient population pharmacokinetics, target organism minimum inhibitory concentration (MIC) distributions, and PK-PD targets from non-clinical models of infection or clinical data have established a new paradigm for determining in vitro susceptibility breakpoints and selection of empirical therapy in clinical practice. Given the increasing problem of antimicrobial resistance, these new tools are valuable additions for clinicians, researchers, and regulatory authorities. PMID- 11050453 TI - Will genomics revolutionize antimicrobial drug discovery? AB - Utilizing genome sequence data from bacterial and fungal pathogens for the discovery of new antimicrobial agents has received considerable attention, both practical and critical, from the pharmaceutical and biotechnological communities. Although no new drugs derived from genomics-based discovery have been reported to be in a development pipeline, the utilization of genomics has revolutionized many aspects of drug discovery. The application, utility, opportunity, and challenges afforded by many of these new approaches are discussed. PMID- 11050455 TI - Making vaccinations easier to swallow. PMID- 11050454 TI - New agents for Gram-positive bacteria. AB - Infections caused by multiple-resistant Gram-positive organisms continue to occur at an alarming rate worldwide. Two new and unique antimicrobial agents targeted specifically against such organisms, quinupristin/dalfopristin and linezolid, have been approved for use in the USA in the past year and will play an important role in the treatment of life-threatening infections. In addition, several new fluoroquinolones have been approved recently or will be available in the near future to aid in the treatment of infections caused by resistant strains of Streptococcus pneumoniae. PMID- 11050456 TI - Crohn's disease treatable in youngsters. PMID- 11050457 TI - Alternative uses found for cancer drug. PMID- 11050458 TI - Drug-excipient interactions and their affect on absorption. AB - Excipient(s) are traditionally thought of as inert but they can have a tremendous impact on the ultimate pharmacological availability of a drug substance when added to a formulation. The magnitude of this effect will depend on the characteristics of the drug and on the quantity and properties of the excipients. The aim of this article is to identify the various physicochemical and physiological processes that can be altered by drug-excipient interactions and to explore mechanisms by which they might occur. The regulatory implications of drug excipient interactions will also be discussed. PMID- 11050459 TI - Enhancing paracellular permeability by modulating epithelial tight junctions. AB - The intestinal epithelium is a major barrier to the absorption of hydrophilic drugs. The presence of intercellular junctional complexes, particularly the tight junctions (zona occludens), renders the epithelium impervious to hydrophilic drugs, which cannot diffuse across the cells through the lipid bilayer of the cell membranes. There have been significant advances in understanding the structure and cellular regulation of tight junctions over the past decade. This article reviews current knowledge regarding the physiological regulation of tight junctions and paracellular permeability, and recent progress towards the rational design of agents that can effectively and safely increase paracellular permeability via modulation of tight junctions. PMID- 11050460 TI - The scope and potential of vaginal drug delivery. AB - The vagina, in addition to being a genital organ with functions related to conception, serves as a potential route for drug administration. Mainly used for local action in the cervico-vaginal region, it has the potential of delivering drugs for systemic effects and uterine targeting. Currently available vaginal dosage forms have several limitations, necessitating the need to develop novel drug delivery systems. In addition, consideration of the regulatory aspects and consumer preferences for vaginal formulations is also required in the early stages of development. PMID- 11050462 TI - Monitor: PROFILE. AB - P-glycoprotein and multidrug resistance-associated protein transporters in the blood-brain barrier: another important brick in the wall PMID- 11050461 TI - Monitor: PROGRESS. AB - Monitor provides an insight into the latest developments in pharmaceutical science and technology through brief synopses of recent presentations, publications and patents, and expert commentaries on the latest technologies. There are two sections: Progress summarizes the latest developments in pharmaceutical process technology, formulation, analytical technology, sterilization, controlled drug delivery systems and regulatory issues; Profiles offers expert commentary on emerging technologies, novel processes and strategic, organizational and logistic issues underlying pharmaceutical R&D. PMID- 11050463 TI - Elevated expression of interleukins in lung adenocarcinomas induced by N Nitrosobis(2-hydroxypropyl)amine in rats. AB - The expression of interleukins (ILs) in lung adenocarcinomas induced by N nitrosobis(2-hydroxypropyl)amine (BHP) in rats was investigated using a multiprobe RNase protection assay (RPA) followed by densitometric quantification. Male Wistar rats, 6 weeks old, were given 2000 ppm BHP in their drinking water for 12 weeks and maintained without further treatment until they were killed at week 25. Total RNAs were extracted from 14 individual adenocarcinomas and 2 specimens of normal lung tissue of untreated rats. In adenocarcinomas, elevated expression of IL-1alpha (6 / 14), IL-1beta (14 / 14), IL-3 (7 / 14), IL-4 (11 / 14), IL-5 (9 / 14), IL-6 (11 / 14) and IL-10 (8 / 14) was observed, compared with normal lung tissues. In contrast, no expression of IL-2 was detected in any case. The results suggest that preferential expression of these ILs and their complex networks may contribute to the development and progression of lung adenocarcinomas induced by BHP in rats. PMID- 11050464 TI - Birth cohort effects on incidence of lung cancers: a population-based study in Nagasaki, Japan. AB - Smoking prevalence remains high (around 60%) among Japanese males, but smoking initiation among males born in the 1930s decreased by approximately 10% due to economic difficulties following World War II. The present study was designed to examine whether this temporary decline in smoking initiation influenced the subsequent incidence of lung cancers, especially adenocarcinoma. Trends of lung cancer incidence by histological type in both sexes were investigated using data from the population-based cancer registry in Nagasaki, Japan, from 1986 through 1995. During this period, 5668 males and 2309 females were diagnosed as having lung cancer, and the overall incidence of lung cancers among both sexes remained stable. However, males aged 55 - 59 years showed a decrease in the age-specific incidence of adenocarcinoma and squamous-cell carcinoma (P < 0.05 and P < 0.01, respectively). In birth cohort analyses, the incidence of adenocarcinoma and squamous-cell carcinoma was lower in the 1935 - 1939 birth male cohort than in the successive cohorts. The incidence of lung cancers among females with low smoking prevalence did not change with birth cohort. The low smoking initiation among the 1935 - 1939 birth male cohort appeared to have resulted in a decreased incidence of adenocarcinoma and squamous cell carcinoma among middle-aged Japanese males. The present study suggests that smoking prevention has an effect in reducing the incidence of lung adenocarcinoma, as well as squamous-cell carcinoma, among smokers. PMID- 11050465 TI - Lack of modification by environmental estrogenic compounds of thyroid carcinogenesis in ovariectomized rats pretreated with N-bis(2 hydroxypropyl)nitrosamine (DHPN). AB - The effects of environmental estrogenic compounds, soy isoflavone mixture (SI), genistein (GEN), and nonylphenol (NP), and the possible goitrogen 3-chloro-4 (dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), on thyroid carcinogenesis were investigated in ovariectomized (OVX) female rats. Five-week-old OVX F344 rats were given a single subcutaneous injection of N-bis(2-hydroxypropyl)nitrosamine (DHPN; 2400 mg / kg, body weight) or vehicle alone. Starting 1 week later, GEN (250 or 25 ppm in diet), SI (400 ppm in diet), NP (250 or 25 ppm in diet), MX (30 ppm, in drinking water), sulfadimethoxine (SDM), a known thyroid tumor-promoter (1000 ppm in drinking water), or beta-estradiol 3-benzoate (EB), a synthetic estrogen (0.5 mg in cholesterol pellet, s.c.) were administered for 12 weeks. SDM and EB were included as positive controls. At sacrifice the major organs including the thyroid, pituitary, liver, kidney, uterus, vagina, brain and pancreas were collected and histopathological observation was performed. Thyroid weights were significantly increased (P < 0. 001) only in the SDM treatment group and pituitary weights were elevated with SDM (P < 0.05) and EB (P < 0.001). Kidney and uterus weights were also significantly increased (P < 0.05) by EB. Histopathologically, proliferative lesions of the thyroid were only observed in the SDM treatment group and of the pituitary in the SDM or EB treatment groups. Renal tubule lesions, uterine squamous metaplasia, vaginal keratinization and telangiectasia of pancreatic islets were also observed with EB. There were no organ weight changes or histopathological lesions in the major organs, including the thyroid, in the GEN, SI, MX or NP treatment groups. Our results thus indicated a lack of modifying effects on thyroid carcinogenesis in female OVX rats, in agreement with our previous finding in males. PMID- 11050466 TI - Increase in 8-hydroxyguanine and its repair activity in the esophagi of rats given long-term ethanol and nutrition-deficient diet. AB - Epidemiological studies have shown that an increased risk of esophageal cancer is associated with the chronic consumption of alcoholic beverages, although alcohol itself is not a carcinogen in animal models. Reactive oxygen species produced by the metabolism of ethanol or by chronic inflammation may play an important role in the carcinogenic process. In this study, we analyzed one type of oxidative DNA damage, 8-hydroxyguanine (8-OH-Gua), and its repair activity in the esophagus as indicators of cellular oxidative stress in rats given long-term ethanol and an autoclaved diet (nutrition-deficient diet). Three-week-old male Sprague-Dawley rats were fed an ethanol beverage whose concentration was increased from 12 to 70% over 20 weeks. When the concentration reached 50%, the diet of one group was changed from the regular diet to an autoclaved diet. At the feeding periods of 20, 25, 30, and 35 weeks, the rats were sacrificed and the 8-OH-Gua levels and repair activities within the esophagi were measured. After 30 weeks of ethanol- and autoclaved diet-feeding, significant increases of 8-OH-Gua and its repair activity were observed in the esophagi, but not in those of the ethanol- and normal diet-fed rats. This result indicates that the combined effects of long term ethanol consumption and nutritional deficiency may be involved in inducing oxidative stress in the rat esophagus. PMID- 11050468 TI - Overexpression of DA41 in v-Ha-ras-3Y1 cells causes growth suppression. AB - We have recently found that DA41 exhibits marked homology with mouse PLIC-1, PLIC 2, frog XDRP1 and yeast DSK2. XDRP1 has been shown to be associated with cyclin A, and blocks cell division of frog embryo. In the present study, we examined the biological role(s) of DA41 in mammalian cells by overexpressing it in v-Ha-ras transformed 3Y1 cells (ras-3Y1). Transfectants which expressed a high level of DA41 mRNA exhibited a decrease in growth rate, a reduction in saturation density, and a suppression of colony formation in soft agar medium. To clarify the molecular mechanism(s) by which DA41 affects cell growth, the effect of DA41 expression on the levels of various cell cycle-regulatory proteins was examined. The forced expression of DA41 gene resulted in a remarkable reduction in CDK2 activity, while the amount of CDK2 did not change. These observations indicate that DA41 is involved in cell cycle regulation in ras-3Y1 cells. PMID- 11050467 TI - Overexpression of midkine in pancreatic duct adenocarcinomas induced by N Nitrosobis(2-oxopropyl)amine in hamsters and their cell lines. AB - The expression of midkine (MK) was investigated in pancreatic ductal hyperplasias, atypical hyperplasias and adenocarcinomas induced by N-nitrosobis(2 oxopropyl)amine (BOP) in hamsters, and in hamster ductal adenocarcinoma cell lines (HPD-1NR, -2NR and -3NR). MK mRNA was clearly overexpressed in invasive pancreatic duct adenocarcinomas (PCs) and the three cell lines as assessed by northern blot analysis, and MK protein expression increased from ductal hyperplasia through atypical hyperplasias, intraductal carcinomas and invasive PCs by immunohistochemistry. The extent of overexpression of MK mRNA in PCs was almost the same as in hamster whole embryonic tissue. MK is reported to be a retinoid-responsive gene, but MK mRNA expression was not affected by treatment with all-trans retinoic acid (tRA) or N-(4-hydroxyphenyl)retinamide (4-HPR) in HPD-1NR cells. The results thus suggest that MK expression is involved in the development and progression of pancreatic ductal adenocarcinomas induced by BOP in hamsters, with loss of upregulation by retinoic acid. PMID- 11050469 TI - Increased frequencies of gene and chromosome mutations after X-irradiation in mouse embryonal carcinoma cells transfected with the bcl-2 gene. AB - Preimplantation stage mouse embryos are known to be highly sensitive to the killing effect of DNA-damaging agents such as radiation. Interestingly, however, this stage of development is well protected from radiation induction of malformation and carcinogenesis in postnatal life. In recent years, it has become clear that the stem cells of preimplantation stage embryos undergo extensive apoptosis after DNA damage. It has been postulated that this apoptosis is likely to be responsible for the resistance to malformation, by excluding cells carrying deleterious DNA damage. We have tested the possible role of apoptosis in elimination of gene and chromosome mutations in undifferentiated mouse embryonal carcinoma cell line, F9, transfected with human bcl-2 cDNA. The colony radiosensitivity of F9 cells was not affected by overexpression of the bcl-2 gene, but the apoptotic cell death was suppressed, as examined by DNA ladder assay and Hoechst staining. This suppression was accompanied by an increase in the frequencies of hprt mutation and micronucleus formation after X-irradiation. These results support the idea that maintenance of genomic integrity during early development is likely to be executed by apoptotic elimination of cells at risk. PMID- 11050470 TI - The specific expression of three novel splice variant forms of human metalloprotease-like disintegrin-like cysteine-rich protein 2 gene inBrain tissues and gliomas. AB - We have previously identified 67 exons on a yeast artificial chromosome contig spanning 1.5 Mb around the multidrug resistance 1 gene region of human chromosome 7q21.1. In this study, we identified three novel cytoplasmic variants (MDC2 gamma, MDC2-delta, and MDC2-epsilon) of the human metalloprotease-like disintegrin-like cysteine-rich protein 2 (MDC2) among these exons by screening a human brain cDNA library and also by using a reverse transcription polymerase chain reaction. Genomic sequence analysis strongly supported the idea that the variations in the cytoplasmic domain were generated by alternative splicing. The expression of MDC2 variant forms in human brain tissue and gliomas was examined by reverse transcription polymerase chain reaction and RNase protection assay. MDC2-epsilon was expressed only in the cortical and hippocampal regions in human brain, but not in gliomas. In contrast, MDC2-gamma was a major form expressed in human gliomas. Specific expression of these cytoplasmic variants of MDC2 in human brain and its malignancies is discussed. PMID- 11050471 TI - Centrosomal kinases, HsAIRK1 and HsAIRK3, are overexpressed in primary colorectal cancers. AB - Members of the recently identified family of Homo sapiens Aurora / Ipl1-related kinases (HsAIRKs), homologous to chromosome segregation kinases, fly Aurora and yeast Ipl1, are highly expressed during M phase, and have been suggested to regulate centrosome function, chromosome segregation, and cytokinesis. In the present study, immunohistochemical analyses were performed of HsAIRK1 and HsAIRK3 expression in 78 primary colorectal cancers and 36 colorectal adenomas as well as 15 normal colorectal specimens. In normal colon mucosa, some crypt cells showed weak positive staining in 10 and 12 out of 15 cases for HsAIRK1 and HsAIRK3, respectively, the remaining cases being negative. Elevated expression of HsAIRK1 was observed in 53 (67.9%) of the colorectal cancers, and of HsAIRK3 in 40 (51.3%). Furthermore, colorectal adenomas showed high expression of HsAIRK1 and HsAIRK3 in 11 (30.6%) and 7 (19.4%) cases, respectively, thus being intermediate between colorectal cancers and normal colorectal mucosa. Interestingly, HsAIRK1 overexpression was significantly associated with pT (primary tumor invasion) and p53 accumulation in colorectal cancers. There was no significant correlation between proliferating cell nuclear antigen-labeling index (PCNA-LI) and the levels of these proteins. The results suggest that overexpression of HsAIRK1 and HsAIRK3 might be involved in tumorigenesis and / or progression of colorectal cancers. PMID- 11050472 TI - Human tissue distribution of TA02, which is homologous with a new type of aspartic proteinase, napsin A. AB - The N-terminal amino acid sequence of TA02 (molecular weight 35.0 kDa, isoelectric point 5.29), which is associated with primary lung adenocarcinoma, was determined and a fragment peptide was used to generate mouse monoclonal antibodies (mAbs) against TA02. The amino acid sequence suggested that TA02 might be homologous with napsin A, a new type of aspartic proteinase. In this context, we confirmed the expression of napsin A in primary lung adenocarcinoma using reverse-transcription polymerare chain reaction (RT-PCR) and showed that the TA02 mAbs reacted with glutathione-S-transferase (GST)-napsin A fusion protein. We concluded that TA02 is the same molecule as napsin A, and showed immunohistochemically that it is distributed mainly in type II pneumocytes, alveolar macrophages, renal tubules and exocrine glands and ducts in the pancreas. In particular, type II pneumocytes and alveolar macrophages showed high expression of TA02 among human normal tissues. In primary lung adenocarcinoma, 47 out of 58 (81.0%) primary lesions were positive. All well-differentiated adenocarcinomas except those of goblet cell type showed high expression of TA02. In addition, two out of seven (28.6%) large cell carcinomas showed low expression of TA02. The other histopathological types of primary lung cancer did not express TA02 at all. A few cases of renal cell cancer, pancreatic cancer, breast cancer, thyroid cancer, colon cancer and ovarian cancer showed low expression, but the staining patterns were completely different from that of primary lung adenocarcinoma, which showed a granular staining pattern. Our novel mAbs should be valuable for immunochemical detection of TA02/napsin A. PMID- 11050473 TI - Activation of intestinal mucosal immunity in tumor-bearing mice by lactoferrin. AB - We have previously demonstrated that oral administration of bovine lactoferrin (bLF) markedly increases CD4(+) and CD8(+) T cells and NK (asialoGM1(+) ) cells in the blood of tumor-bearing mice and enhances anti-metastatic activity. In this paper, we document that oral administration of bLF and bLF-hydrolysate (bLFH) is associated with strong increases in CD4(+) and CD8(+) T, as well as asialoGM1(+) cells in lymphoid tissues and lamina propria of the small intestine in mice, especially in tumor-bearing animals in which Co26Lu cells were implanted subcutaneously. Moreover, IgM(+) and IgA(+) B cells in lamina propria of the small intestine were also significantly increased by bLF and bLFH. Bovine apo transferrin (bTF) did not exhibit such activity. In the colon, only CD8(+) cells were significantly increased by treatment with bLF, while asialoGM1(+) cells were significantly decreased. bLF and bLFH induced cytokines to activate T, B and asialoGM1(+) cells. Administration of bLF and bLFH, but not bTF, increased production of interleukin-18 (IL-18), interferon-gamma (IFN-gamma) and caspase-1 in the mucosa of the small intestine. Particularly high levels of IL-18 were found in the epithelial cells of the small intestine. Moreover, administration of bLF and bLFH, but not bTF, induced IFN-gamma presenting cells in the small intestine. Caspase-1, which processes proIL-18 to mature IL-18, was also induced in the epithelial cells of the small intestine following treatment with bLF and bLFH, but not with bTF. These results suggest that enhanced production of IL-18 and IFN-gamma and caspase-1 induction by treatment with bLF may be important for elevation of intestinal mucosal immunity. PMID- 11050474 TI - Transduction of a fiber-mutant adenovirus for the HSVtk gene highly augments the cytopathic effect towards gliomas. AB - Suicide gene therapy utilizing the herpes simplex thymidine kinase (HSVtk) / ganciclovir (GCV) system has been performed to kill cancer cells. However, the low transduction efficiency of HSVtk gene into cancer cells critically limits its efficacy in cancer treatment in clinical situations. To improve delivery of the HSVtk gene into cancer cells, we transduced U-87MG and U-373MG glioma cells with adenovirus (Adv) vectors with a fiber mutant, F / K20, which has a stretch of 20 lysine residues added at the C-terminus of the fiber, for the HSVtk gene (Adv-TK F / K20), and compared the cytopathic effect of Adv-TK-F / K20 with that of the Adv for HSVtk with wild-type fiber (Adv-TK). The cytopathic effect of Adv-TK-F / K20 in U-87MG and U-373MG cells was approximately 140 and 40 times, respectively, stronger than that of Adv-TK. At the same multiplicity of infection (MOI) in each cell line, Adv-TK-F / K20 induced a higher degree of apoptosis (U-87MG, 35%; U 373MG, 77%) than Adv-TK (U-87MG, 0.11%; U-373MG, 27%) in U-87MG (MOI 0.03) and U 373MG cells (MOI 0.1). Cleavage of poly(ADP-ribose)polymerase (PARP) was more marked in the cells that were infected with Adv-TK-F / K20 than in cells that were infected with Adv-TK. These results indicate that gene therapy utilizing Adv TK-F / K20 may be a promising therapeutic modality for the treatment of gliomas. PMID- 11050475 TI - Production of a single-chain variable fragment antibody recognizing type III mutant epidermal growth factor receptor. AB - The type III deletion mutant of the epidermal growth factor receptor (EGFR) is a potential target in diagnostic and therapeutic approaches for those glioblastomas characterized by its expression. We previously raised a mouse monoclonal antibody, 3C10 (IgG2b) specifically recognizing this mutant EGFR. In this study, a single-chain variable fragment (scFv) antibody was produced. Partial determination of its N-terminal amino acid sequence and preparation of adequate primers for variable heavy chain (V(H)) and variable light chain (V(L)) genes were performed to allow cloning by means of reverse transcriptase-polymerase chain reaction. The genes cloned were assembled with a linker, (Gly4Ser)3, and ligated into a bacterial expression vector to express the scFv as cytoplasmic inclusion bodies. After appropriate refolding, the antibody activity of the V(H) V(L) scFv was examined in an enzyme-linked immunosorbent assay. 3C10 scFv showed a selective reactivity with the mutant peptide, similarly to the parental 3C10 antibody. A mouse transfectant expressing the type III mutant EGFR and a glioblastoma with type III deletion-mutant EGFR were positively stained by immunofluorescence. By Biacore analysis, the affinity (K(A)) of the parental 3C10 for the mutant peptide was 9.7 x 10(7) M(-1), while that of 3C10 scFv was 2.45 - 2.48 x 10(7) M(-1), being approximately 4-fold weaker. The results together suggested that the scFv antibody retained the appropriate structure to recognize a conformational epitope of the mutant receptor, similarly to the parental antibody. PMID- 11050476 TI - Adenovirus-mediated transfer of Fas ligand gene augments radiation-induced apoptosis in U-373MG glioma cells. AB - Most malignant astrocytomas (gliomas) express a high level of Fas, whereas the surrounding normal tissues such as neurons and astrocytes express a very low level of Fas. Thus, transduction of Fas ligand would selectively kill malignant astrocytoma cells. On the other hand, glioma cells harboring p53 mutation have been reported to be resistant to conventional therapies including radiation. To override the resistance mechanism of glioma cells with p53 mutation to radiation, we transduced U-373MG malignant astrocytoma (glioma) cells harboring mutant p53 with Fas ligand via an adenovirus (Adv) vector in combination with X-ray irradiation, and evaluated the degree of apoptosis. The degree of apoptosis in U 373MG cells infected with the Adv for Fas ligand (Adv-FL) and treated with irradiation (81%) was much higher than that in U-373MG cells infected with Adv-FL and not treated with irradiation (0.8%) or that in U-373MG cells infected with the control Adv for lacZ and treated with irradiation (5.0%). In U-373MG cells infected with Adv-FL, irradiation increased the expression of Fas ligand. Coincident with the increase in Fas ligand, there was a marked reduction in the caspase-3 level and a marked increase in the cleaved form of poly(ADP-ribose) polymerase (PARP), which are downstream components of Fas ligand-mediated apoptosis. This suggests that the enhanced activation of caspase-3 by the transduction of Fas ligand combined with irradiation, induced extensive apoptosis in U-373MG cells. In summary, transduction of Fas ligand may override the resistance mechanism to radiotherapy in glioma cells harboring p53 mutation. PMID- 11050478 TI - The effects of boron neutron capture therapy on liver tumors and normal hepatocytes in mice. AB - To explore the feasibility of employing boron neutron capture therapy (BNCT) to treat liver tumors, the effects of BNCT were investigated by using liver tumor models and normal hepatocytes in mice. Liver tumor models in C3H mice were developed by intrasplenic injection of SCCVII tumor cells. After borocaptate sodium (BSH) and boronophenylalanine (BPA) administration, (10)B concentrations were measured in tumors and liver and the liver was irradiated with thermal neutrons. The effects of BNCT on the tumor and normal hepatocytes were studied by using colony formation assay and micronucleus assay, respectively. To compare the effects of BSH-BNCT and BPA-BNCT, the compound biological effectiveness (CBE) factor was determined. The CBE factors for BSH on the tumor were 4.22 and 2.29 using D(10) and D(0) as endpoints, respectively. Those for BPA were 9.94 and 5.64. In the case of hepatocytes, the CBE factors for BSH and BPA were 0.94 and 4.25, respectively. Tumor-to-liver ratios of boron concentration following BSH and BPA administration were 0.3 and 2.8, respectively. Considering the accumulation ratios of (10)B, the therapeutic gain factors for BSH and BPA were 0.7 - 1.3 and 3.8 - 6.6, respectively. Therefore, it may be feasible to treat liver tumors with BPA-BNCT. PMID- 11050477 TI - Association of preoperative radiation effect with tumor angiogenesis and vascular endothelial growth factor in oral squamous cell carcinoma. AB - This study examined the relationship between tumor angiogenesis and the radiation induced response, evaluated based on pathological changes, in oral squamous cell carcinoma patients treated with preoperative radiation therapy. Forty-one cases of squamous cell carcinoma treated with preoperative radiation therapy were investigated. Tumor angiogenesis was assessed by scoring the intratumor microvessel density (IMVD). Expression of vascular endothelial growth factor (VEGF) was also evaluated before and after preoperative radiotherapy. There was no correlation between IMVD in the specimens before therapy and the pathological response to radiation therapy. However, radiation therapy decreased IMVD in the specimens after therapy. A significant association was observed between VEGF expression and resistance to radiation therapy: only 4 of the 21 patients whose tumors exhibited a high level (2 + or 3 + ) of VEGF staining experienced a major (3 + or 4 + ) pathological response to radiation therapy. Furthermore, an increasing level of VEGF expression after radiation therapy was observed in non effective (0 to 2 + ) response cases. These results suggest that VEGF expression and the induction of this protein are related to radiosensitivity and could be used to predict the effects of preoperative radiation therapy on oral squamous cell carcinoma. PMID- 11050479 TI - Mechanism of anti-tumor effect of combination of bleomycin and shock waves. AB - We have previously reported marked enhancement of the cytocidal effect of bleomycin (BLM) on cancer cell suspensions in vitro by the combination with shock waves. In this study, we evaluated the synergistic effects on cancer cell proliferation and apoptosis in solid tumors. A spherical piezo-ceramic element was used as the shock wave source, with a pressure peak of 40 MPa. A human colon cancer cell line, SW480 was implanted onto the back of nude mice. Two thousand shock waves were administered to the tumor immediately following an intravenous injection of BLM at a dose of one-tenth of the LD(50). The tumor was extirpated at 3, 6, 12, 24, 72 h and 1 week following shock exposure. Cell proliferation and apoptosis were detected by Ki-67 using antibody MIB-1 and by the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-biotin nick-end labeling (TUNEL) method. The lowest percentage (35.7%) of Ki-67-positive cells appeared 24 h following the treatment. The maximum apoptotic index was detected within 6 h following the treatment. Moreover, numerous large cells with enlarged nuclei were detected histologically. These results suggest that shock waves may enhance chemotherapeutic effects by increasing apoptosis and decreasing cell proliferation in the tumor tissue. PMID- 11050481 TI - Treatment of Helicobacter pylori: an overview. AB - Helicobacter pylori is recognized to be a serious pathogen, but there is still controversy as to who should be treated. There is consensus for treatment of Helicobacter-positive peptic ulcer and B-cell lymphoma. Lymphocytic gastritis and giant-fold gastritis (Menetrier's disease) may also respond to treatment. Patients with func-tional dyspepsia have a 20% placebo response with a 5-10% 'eradication' response, results not dissimilar from empirical treatment with a proton pump inhibitor. A 'test and treat' policy for patients with uninvesti gated dyspepsia remains controversial. Some have suggested that eradication may increase the risk of GERD, or predispose to adenocarcinoma at the gastro oesophageal junction. However, PPI treatment without Helicobacter eradication induces greater inflammation in the gastric corpus, the phenotype associated with non-cardia gastric cancer. A minority believe that Helicobacter should be eradicated in all individuals. When choosing treatment it is logical to start with a combination of antibiotics that, in the event of failure, will allow a second combination to be used without overlap. PMID- 11050480 TI - Structure-activity relationship of a novel group of mammalian DNA polymerase inhibitors, synthetic sulfoquinovosylacylglycerols. AB - We reported previously that sulfolipids in the sulfoquinovosylacylglycerol class from a fern and an alga are potent inhibitors of DNA polymerase alpha and beta and potent anti-neoplastic agents. In developing a procedure for chemical synthesis of sulfolipids, we synthesized many derivatives and stereoisomers of sulfoquinovosylmonoacylglycerol (SQMG) / sulfoquinovosyldiacylglycerol (SQDG). Some of these molecules were stronger inhibitors than the SQMG / SQDG originally reported as natural compounds. In this study, we examined the structure inhibitory function relationship of synthetic SQMG / SQDG and its relationship to cytotoxic activity. The inhibitory effect is probably mainly dependent on the fatty acid effect, which we reported previously, although each of the SQMG / SQDG was a much stronger inhibitor than the fatty acid alone that was present in the SQMG / SQDG. The inhibitory effect could be influenced by the chain size of fatty acids in the SQMG / SQDG. The sulfate moiety in the quinovose was also important for the inhibition. Lineweaver-Burk plots of SQMG / SQDG indicated that DNA polymerase alpha was non-competitively inhibited, but the SQMG / SQDG were effective as antagonists of both template-primer DNA-binding and nucleotide substrate-binding of DNA polymerase beta. The SQMG had an cytotoxic effect, but the SQDG tested did not. The SQDG might not be able to penetrate into cells. Based on these results, we discuss the molecular action of SQMG / SQDG and propose drug design strategies for developing new anti-neoplastic agents. PMID- 11050482 TI - Review article: have we found the source of Helicobacter pylori? AB - Besides the well established Helicobacter pylori reservoir, i.e. the human stomach, numerous other sources have been hypothesized. However, none has been definitely proven. In some instances (pig, sheep), Helicobacter species closely related but different from H. pylori were detected but the results were misleading because culture of sufficiently discriminating molecular techniques were not used. In other cases, the strain was really H. pylori (cat) but the case was anecdotal or the animal species (monkey) has so little contact with humans that the possible source has no epidemiological consequence. This is also the case for houseflies which theoretically can be a vehicle, but practically speaking are not because of too few viable bacteria present in faeces. Molecular epidemiology studies demonstrating the route of transmission (faecal-oral, oral oral or gastro-oral) are still lacking but recent studies have confirmed the presence of viable H. pylori in vomitus and in faeces in the event of diarrhoea. PMID- 11050483 TI - Review article:invasive and non-invasive tests for Helicobacter pylori infection. AB - There are two general ways in which a diagnosis of infection by Helicobacter pylori can be made: by using either an invasive or non-invasive procedure. The invasive procedures involve an endoscopy and biopsy. A biopsy is essential because often the mucosa may appear macroscopically normal but nevertheless be inflamed. A biopsy is obtained by histological examination, culture, polymerase chain reaction or detection of the presence of urease activity in biopsy material. The non-invasive tests that can be used to diagnose the infection are serology, detection of labelled metabolic products of urea hydrolysis in the breath (13CO2, 14CO2), the urine or the blood, and detection of H, pylori antigen in a stool specimen. At present no single test can be relied upon to detect definitely colonization by H. pylori, and a combination of two is recommended if this is feasible. The choice of the test to be used is not straightforward and may vary according to the clinical condition and local expertise. PMID- 11050484 TI - Review article: the continuing dilemma of dyspepsia. AB - Dyspepsia drains a substantial proportion of healthcare resources in industrialized countries and an appropriate management strategy is needed. An aetiological role for Helicobacter pylori infection has been demonstrated in a number of pathological conditions associated with dyspepsia, such as peptic ulcer and gastric malignancies, but not in functional dyspepsia. Endoscopy and diagnosis-based treatment, H. pylori testing and eradication therapy, history taking and empirical therapy, are the main tools that are currently available for managing patients with upper gastrointestinal symptoms. Endoscopy identifies malignancies and organic diseases of the proximal gut and therefore provides reassurance to both doctors and patients. It should be recommended in older patients with suspicious symptoms and it has proven to be more cost-effective than empirical H2-receptor antagonists in patients with ulcer-like symptoms. Empirical eradication in all dyspeptics without suspicious symptoms is a cost effective approach that cures the majority of peptic ulcers. Nevertheless, it does not control symptoms in the majority of patients, it may exacerbate gastro oesophageal reflux disease, and it encourages antibiotic resistance. The realities of current clinical practice require empirical therapy in most, if not all, the dyspeptics seen by general practitioners. A detailed history taking can help to diagnose gastro-oesophageal reflux disease and to identify suspicious symptoms. Furthermore, identification of dyspepsia subgroups may provide guidance for empirical therapy. Nevertheless, even analysis of individual symptoms does not provide a sufficient diagnostic yield to differentiate functional from organic dyspepsia and appropriate investigations are needed in patients with poor response to short-term therapy or frequent relapses. PMID- 11050485 TI - Review article: is Helicobacter pylori status relevant in the management of GORD? AB - There is growing interest in the relationship between H. pylori infection and gastro-oesophageal reflux disease (GORD). However, this relationship is complex, as yet not fully elucidated, and probably based on a multiplicity of factors. The prevalence of H. pylori infection in patients with GORD is similar, more often lower than in matched controls. There is a negative correlation between H. pylori infection and the severity of GORD. There are many hypothetical mechanisms by which H. pylori infection may protect from the development of GORD. Conversely, there are many possible mechanisms by which H. pylori infection could theoretically foster the GORD. Patients after H. pylori eradication may develop GORD, and this seems to suggest a protective role of H. pylori infection, but other possible explanations include weight gain after H. pylori eradication, changes in dietary habits and smoking, and pre-existing GORD. H. pylori infected patients treated by various acid-inhibiting therapies such as proton pump inhibitors (PPIs), H2-receptors antagonists (H2-RA) or vagotomy, have an increase of their corpus gastritis severity, both in the activity of inflammation and in the density of organisms. Long-term therapy of GORD in H. pylori infected may lead to rapid progression of atrophic gastritis intestinal metaplasia and dysplasia, and increase the risk of developing gastric cancer. More recently it has been shown that H. pylori infection may interfere with the acid suppressive therapies used for treating GORD. In our opinion the progression of gastritis depends on the threshold of acid output at which H. pylori can 'flourish'. Recently interest is growing on gastric transitional zones and Helicobacter ecology. Any decrease of acid secretion changes the behaviour of H. pylori: the activity of gastritis improves in the antrum, but it deteriorates in the body. During proton pump inhibitor treatment, H. pylori redistribution occurs within the stomach, from an antral to a corpus or fundus prevalent pattern; corpus fundus gastritis, exacerbated by PPI therapy, may result both in a diminished acid secretion and gastro-oesophageal reflux. The interest in Barrett's oesophagus is growing due to the associated risk of adenocarcinoma. The literature seems to demonstrate that the prevalence of H. pylori infection of the stomach in Barrett's oesophagus patients is not different from that exhibited by controls, roughly one-third of the subjects. Intestinal metaplasia of the gastric cardia seems to be equally frequent in patients with and without GORD. Finally, it appears unlikely that a causal relationship exists between H. pylori infection and Barrett's-associated adenocarcinoma. PMID- 11050486 TI - Review article: non-steroidal anti-inflammatory drugs and Helicobacter pylori. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) and Helicobacter pylori are well recognized causes of gastroduodenal mucosal damage. This damage is mediated through the effects of both agents on acid secretion, neutrophil activity and function, and prosta- glandin metabolism. Clinical trials on the interrelation- ship between H. pylori, NSAIDs and gastroduodenal mucosal injury have yielded conflicting results. No consensus has been reached on what recommenda- tions should be implemented with regard to H. pylori eradication in patients on long term NSAID therapy. At present, the presence of H. pylori is identified at endoscopy and eradication is carried out in symptomatic patients. Asymptomatic patients remain a dilemma that requires further investigation. Clinical practice will continue to be tailored to a patient's individual requirements. Therefore, in patients at risk of gastrointestinal haemorrhage, and on NSAID therapy, acid suppression therapy should be prescribed. PMID- 11050487 TI - Review article: Helicobacter pylori and gastric cancer--the clinicians'point of view. AB - Although the incidence of gastric cancer has declined dramatically in Western countries, the most recent data from the International Agency for Research on Cancer show that it remains the second most common cancer worldwide and caused 628 000 deaths in 1990. The incidence and prevalence of gastric cancer are projected to increase over the next few decades in less developed countries as a result of the increased longevity of H. pylori-infected populations and improved therapies. Gastric carcinogenesis is a multistep and multifactorial process beginning with H. pylori-associated gastritis in most cases. H. pylori infection, together with other environmental factors and individual susceptibility, determine the final risk for the development of gastric cancer. The magnitude of H. pylori infection as a risk factor for gastric cancer in the published H. pylori and gastric cancer epidemiology studies may have been underestimated due to the inclusion of improperly selected controls. Eradication of the infection has been shown to prevent the occurrence of metachronous gastric cancer following endoscopic resection of early gastric cancer in a Japanese study. However, the generalization of this study to other populations is difficult because of the vast differences in the definition of gastric atrophy and early gastric cancer between Japanese and Western pathologists. Until an international consensus on the pathological diagnosis of gastric atrophy and early gastric cancer is reached, interpretation of studies performed in different countries remains difficult. Clinicians rely on the correct pathological diagnosis to guide the management of H. pylori infection-associated gastrointestinal diseases. PMID- 11050488 TI - Review article: Helicobacter pylori: where are we and where are we going? AB - This paper contains a personal view on what has been achieved in Helicobacter pylori research and what the expectations might be for further developments. Knowledge about the organism is already extensive. Particularly intriguing are the differences in genetic make-up in the various geographical regions. Sadly, detailed knowledge on how the organism spreads is still lacking. The clinical spectrum of the disease in man is largely known, but as H. pylori is disappearing worldwide, the relative frequency of H. pylori-negative ulcer disease is increasing. To what extent H. pylori disappearance and eradication is responsible for the decreasing incidence of gastric cancer remains uncertain. Antimicrobial therapy is dominated by proton pump inhibitor triple therapy as first line with quadruple therapy as second-line therapy. The long-term consequences of the rising resistance to the 'key' antimicrobials are so far unknown and speculative. PMID- 11050489 TI - A systematic review of Helicobacter pylori eradication treatment schedules in children. AB - METHODS: We systematically reviewed all available data in the paediatric literature on treatment for Helicobacter pylori infection to determine overall efficacy of different schedules. A comprehensive search of all published articles and letters from 1987, and of abstracts presented at three main meetings on this topic between 1997 and 1999, was carried out. Results from all English and French papers, letters and abstracts were extracted and analysed. RESULTS: Only 30 full articles and 16 abstracts were found, with results on eradication of H. pylori in 870 and 1552 children, respectively. Monotherapy or dual therapy with an antisecretory drug plus one antibiotic showed a very low efficacy. Dual therapies with bismuth plus one antibiotic (either amoxycillin or a nitro- imidazole) or two antibiotics when administered for 2 or more weeks were as effective as either bismuth-based or proton pump inhibitor-based triple therapies. Triple therapies were less effective than in adults, and while bismuth-based triple therapies were more effective when given for 2 weeks than for one week, proton pump inhibitor based triple therapies have a similar efficacy irrespective of the duration. CONCLUSION: In children dual therapies seemed as effective as triple therapies, and longer courses of proton pump inhibitor-base triple therapies are not better than shorter ones. PMID- 11050490 TI - The history of electrosurgery. PMID- 11050491 TI - Mohs surgery and processing: novel optimizations and enhancements. AB - OBJECTIVE: Simple yet effective modifications to Mohs surgery and processing may enhance procedural efficiency, ensure proper tissue orientation and tracking, while greatly reducing "recuts." Using the methods described, Mohs specimens no longer need to be incised or excised with any bevel, thus conserving tissue and facilitating closure. METHODS: A streamlined Mohs surgical tray is convertible to a closure tray within seconds. The excised tissue specimen is oriented on a sterile paper square on a reusable sterilized aluminum palette where partial thickness circumferential and radial scalpel cross-hatching allows epidermal edges complete freedom to later adhere to a flattening glass. The sterile paper can be labeled with patient name, stage number, and chuck number; then the specimen is inked. Rapid chuck freezing in a specially positioned liquid nitrogen immersion is followed by OCT (embedding compound) application. Uniquely numbered and modified cryostat chucks eliminate the possibility of OCT-chuck disunion. Rapid liquid nitrogen immersion of a glass surface allows the inked, cross hatched specimen's epidermal edges and base to lay perfectly flat once forced against the supercooled glass surface using a special polymer glove. Inversion of the specimen-containing glass onto a frozen and gel state OCT interface of the chuck completes the embedding. RESULTS/CONCLUSION: These reproducible approaches to Mohs surgery described herein utilize multiple modifications that enhance the speed, efficiency, and reproducibility of Mohs specimen embedding, specimen preparation, while maintaining accuracy of interpretation. PMID- 11050492 TI - Full-face nonablative dermal remodeling with a 1320 nm Nd:YAG laser. AB - BACKGROUND: Full-face laser-induced dermal remodeling has traditionally involved ablative methods with their associated complications and limitations. Rhytide improvement requires, among other things, dermal collagen remodeling. Such remodeling has been shown to occur without the requirement of epidermal ablation. OBJECTIVE: To evaluate the investigator's objective and patient's subjective improvement seen after full-face treatment with a 1320 nm Nd:YAG laser. METHODS: Ten patients with class I-II rhytides and Fitzpatrick skin types I-II were treated five times over 3-4 week intervals with a 1320 nm Nd:YAG laser. Patients were evaluated for degree of clinical improvement 6 months after their final treatment. RESULTS: All 10 subjects reported subjective improvement in the quality of their skin. Only six subjects were felt by the investigator to show definitive clinical improvement. Six-month posttreatment biopsies showed evidence of new collagen formation. CONCLUSION: Irradiation with 1320 nm Nd:YAG laser can lead to new collagen formation and associated clinical improvement. Such full face improvement can occur without epidermal ablation. PMID- 11050493 TI - An in vivo trial comparing the clinical efficacy and complications of Q-switched 755 nm alexandrite and Q-switched 1064 nm Nd:YAG lasers in the treatment of nevus of Ota. AB - BACKGROUND: Q-switched 755 nm alexandrite (QS alex) and Q-switched 1064 nm Nd:YAG lasers are effective in the treatment of nevus of Ota. Our previous in vivo study indicated that patients better tolerate QS alex than QS 1064 nm Nd:YAG laser. However, in terms of clinical efficacy and long-term complications, the study did not indicate which laser is superior. Although both machines may appear to be similar in effectiveness, the low number of treatment sessions may contribute to this apparent lack of difference. OBJECTIVE: The aim of this study was to compare the clinical efficacy and complications of QS 755 nm alex and QS 1064 nm Nd:YAG lasers in the treatment of nevus of Ota after three or more treatment sessions. METHOD: Forty patients were recruited for this study and all had received three or more laser treatment sessions with an interval of at least 2 months between each. Half of the lesion was treated with QS alex and the other half with QS 1064 nm Nd:YAG laser. The degree of lightening was assessed subjectively by the patient using a visual analog scale and objectively by two independent clinicians. Patients were called back to be examined for evidence of complications. RESULTS: In terms of subjective degree of lightening, QS 1064 nm Nd:YAG laser was found to be significantly more efficacious than QS alex (P = 0.018). Both clinicians also found QS 1064 nm Nd:YAG laser to be more effective, but statistical significance was only detected in one, not both of their scores (P = 0.005 and 0.414 for observers 1 and 2, respectively). More patients that received QS Alex developed complications (4 for QS alex and 2 for QS Nd:YAG), but the difference was not statistically significant. CONCLUSION: QS 1064 nm Nd:YAG laser appears to be more effective than QS alex in the lightening of nevus of Ota after three or more laser treatment sessions. However, the majority (55%) of the patients reported no differences in results between the two lasers, and only one of the two observers noted statistically significant improvement of QS 1064 nm Nd:YAG over QS alex. PMID- 11050494 TI - Conscious sedation in dermatologic surgery. PMID- 11050495 TI - Tacky but refined: a "slick" technique for dressings that hold better. AB - For decades benzoin has been used to enhance the "stickiness" of the skin for improved adherence complementing the protective and functional aspects of well constructed surgical bandages/dressings. The fully adherent and longer-lasting pressure dressing is both aesthetically pleasing to the patient (and surgeon) and, at the same time, functions to decrease seroma or hematoma formation, enhances antisepsis by blocking transient bacterial contamination, and inhibits traumatic sabotage of surgical wounds due to patient overactivity or inadvertent collision with a door jam, car door, table leg, or grandchild, etc. Also, when the bandage looks good, the patient realizes the operation was a success (at least aesthetically). Although simple, our additional steps of deoiling the skin with acetone, followed by multiple compress/release cycles of gauze to the Mastisol area reaps benefits to the patient and surgeon both in function (adherence) and in the production of a truly "dressy" beautiful and long-lasting dressing. In this way, the dressing "bed" is not only more receptive to one's aesthetically pleasing bandage, but is more tacky, yet refined. This same technique can be utilized prior to application of adhesive strips to healing wounds after sutures have been removed, allowing them to "stay on" longer with potentially better cosmesis and less spreading of the scar. PMID- 11050496 TI - Congenital spitz nevus. AB - BACKGROUND: Congenital Spitz nevus has been reported previously in the literature, but the histopathologic features have not been examined in detail. OBJECTIVE: To histologically examine and report on congenital Spitz nevus. METHOD: We examined 10 clinically submitted congenital melanocytic nevi that were histopathologically identified as congenital Spitz nevi and compared them to the characteristics seen in acquired Spitz nevus and superficial congenital melanocytic nevus. RESULTS: Of the 10 congenital Spitz nevi, 9 were compound and 1 was dermal. Two showed features of combined Spitz nevus (Spitz and blue). Six cases showed all 16 listed characteristics of acquired Spitz nevus, with two cases having 15 and two cases having 14 characteristics. Of the superficial congenital melanocytic nevus characteristics, all except three cases had all 12 attributes. The one dermal lesion had all the characteristics of the acquired Spitz nevus and all but one of the characteristics of the superficial congenital melanocytic nevus in regards to intradermal findings. CONCLUSIONS: Congenital Spitz nevi are true congenital lesions, with histopathologic features of both acquired Spitz nevus and superficial congenital melanocytic nevus. PMID- 11050497 TI - Self-limited adverse reaction to human-derived collagen injectable product. AB - BACKGROUND: Soft tissue augmentation is a common and safe cosmetic and reconstructive procedure. OBJECTIVE: We describe a temporary and self-limited adverse reaction to Dermalogen. METHODS: Clinical and histologic evaluation following an adverse reaction noted at a Dermalogen skin test site. RESULTS: Our patient was found to have a foreign body reaction to Dermalogen. CONCLUSION: Dermalogen, a form of acellular human collagen, may induce a foreign body reaction. PMID- 11050498 TI - Kaposi's varicelliform eruption in a patient with healing peribucal dermabrasion. AB - Kaposi varicelliform eruption (KVE) is the name given to a distinct cutaneous eruption caused by Herpesvirus hominis types 1 and 2, vaccinia virus, or coxsackie A16 virus, superimposed on a preexisting dermatosis. A delay in diagnosing this condition may result in intense pain and rapid spread of the cutaneous lesions. We report a patient who underwent perioral dermabrasion for wrinkles who developed KVE secondary to herpes simplex virus infection. PMID- 11050499 TI - Dermatofibrosarcoma protuberans growing around plantar aponeurosis: excision by Mohs micrographic surgery. AB - BACKGROUND: Dermatofibrosarcoma protuberans (DFSP) is a spindle cell malignancy that has a high local recurrence rate after excision with minimal or no immediate tissue margin assessment. DFSP is exceedingly rare on the palms and soles. OBJECTIVE: To report a case of a locally aggressive DFSP on the sole excised using Mohs micrographic surgery. METHODS: Case report and review of the literature. RESULTS: Mohs micrographic surgery unmasked tumor infiltration that extended around plantar aponeurosis and into underlying plantar muscle fascia. CONCLUSION: Mohs micrographic surgery should be considered the treatment of choice for DFSP, especially in acral locations. This technique allows the surgeon to trace out deep tumor extensions that may wrap around underlying tendon, a finding that may not be appreciated clinically. PMID- 11050500 TI - Graftskin heals an ulcer on an amputation stump. AB - BACKGROUND: A 51-year-old man presented with a chronic ulcer of 28 years' duration on the amputation stump of his right foot. The prosthesis was aggravating the ulcer, despite multiple therapies. OBJECTIVE: To heal the ulcer and avoid a below-the-knee amputation. Graftskin, a bilayered living skin construct, was applied. METHODS: Graftskin was sutured in place following gentle debridement. Pentoxifylline was given to improve circulation. The dressing was changed 5 days after Graftskin application, then weekly for 7 weeks. RESULTS: Crusting was evident at 3 weeks, with an underlying white coating at 4 weeks. By 7 weeks, full reepithelialization was present under the crust and no ulceration remained. Over succeeding weeks the ulcer bed became flush with the surrounding skin. The patient's pain ceased, and some sensation and warmth returned to the stump. CONCLUSION: Graftskin treatment successfully healed a chronic ulcer, possibly preventing a more extensive amputation. PMID- 11050501 TI - Resurfacing extensive malar and preauricular cutaneous defects with pedicled temporoparietal fascia. AB - BACKGROUND: The pedicled temporoparietal fascial flap (TPFF) is useful for preparing extensive cutaneous facial defects for skin grafting. OBJECTIVE: The purpose of this work is to describe the use of the pedicled TPFF for preparation of extensive cutaneous malar and preauricular facial defects for skin grafting and contour improvement. METHODS: Two patients underwent transposition of a TPFF and immediate skin grafting. Patient demographics, diagnoses, wound characteristics, and outcome are described and documented through photographs. RESULTS: The TPFF greatly simplified reconstruction in the cases presented. The TPFF provided a vascularized tissue bed that resulted in complete survival of the skin grafts. It also effectively replaced lost volume and corrected contour deformities. CONCLUSION: The use of the pedicled TPFF for extensive malar and preauricular cutaneous defects provides a vascular tissue bed for skin grafting and soft tissue replacement. The TPFF may obviate the need for more complicated and morbid reconstructive procedures. PMID- 11050502 TI - Myiasis in a pregnant woman and an effective, sterile method of surgical extraction. AB - BACKGROUND: Cutaneous myiasis caused by Dermatobia hominis involves the infestation of tissue with dipterous fly larvae and is common in many tropical and subtropical areas. We describe a patient in her sixth month of pregnancy who returned from Belize with myiasis detected in the right popliteal fossa. Multiple surgical techniques have been described in the past outlining various ways to extract the botfly larva. No single standardized technique for surgical extraction of larvae has been adopted. OBJECTIVE: To describe a highly effective, sterile method of extraction used in a pregnant patient with botfly infestation. METHODS: A combination of injection with plain 1% lidocaine, sterile occlusion with polymyxin B sulfate ointment and a cruciform incision was used to extract the larva. RESULTS: The botfly larva was easily and completely extracted without remnants of the larval body being left in the skin. CONCLUSION: The sterile technique we describe allows for quick and easy extraction of the larva without risk of secondary infection or need for antibiotics. This method is especially appropriate for pregnant patients or those with medical conditions precluding a completely competent immune response to potential secondary infection. PMID- 11050503 TI - Treatment of lichen amyloidosis (LA) and disseminated superficial porokeratosis (DSP) with frequency-doubled Q-switched Nd:YAG laser. AB - BACKGROUND: Under normal practice, the full thickness of the epidermis is peeled off when treating pigmented lesions with a frequency-doubled Nd:YAG laser. Based on this observation, it is postulated that this laser may be effective for treating lichen amyloidosis (LA) and disseminated superficial porokeratosis (DSP) for which the pathologic changes are limited to the epidermis and papillary dermis. OBJECTIVE: To investigate the clinical effect of frequency-doubled Nd:YAG laser treatment for one patient with LA and for one patient with DSP. METHODS: Frequency-doubled 532 nm Nd:YAG laser pulses were delivered to the lesions on the limbs of a patient with LA and the face and forearms of a patient with DSP. Lesions of LA were treated two or three times, and those of DSP were treated four times, treatment sessions being 1 month apart. For this investigation, biopsies were taken from untreated lesions prior to treatment, lesions immediately after laser treatment, and lesions present at a 9-month follow-up investigation. RESULTS: Both the patient with LA and the patient with DSP responded well to treatment, the results of which remained unchanged at a follow-up conducted 9 months after the final treatment session. CONCLUSION: Frequency-doubled Nd:YAG laser treatment provided excellent results for the patient with LA as for the patient with DSP. The effectiveness of this method deserves further study in a larger group of test patients. PMID- 11050504 TI - Liposuction Council bulletin. PMID- 11050505 TI - Reconstruction conundrum #3. Excision and reconstruction of recurrent lentigo maligna melanoma. PMID- 11050507 TI - Boxer shorts, gummy bears and lasered hairs. PMID- 11050506 TI - Alar batten cartilage grafting. PMID- 11050508 TI - Advertising & the use of the media in dermatology--this is appropriate in the new millennium. PMID- 11050509 TI - Lessons on dermoscopy: case #9. Congenital melanocytic nevus. PMID- 11050510 TI - Perpendicular suture method for earlobe keloids. PMID- 11050511 TI - More on Monsel's solution. PMID- 11050512 TI - Goat lab as alternative or adjunct to pig head model for practice cutaneous surgery. PMID- 11050513 TI - Regarding the modified Burow's wedge flap for upper lateral lip defects. PMID- 11050514 TI - Inadvertant and undesirable sequelae of the stellate purse-string closure. PMID- 11050515 TI - Regarding the youthful forehead: placement of skin incisions in hidden furrows and a poor esthetic result. Something else must have happened! PMID- 11050516 TI - Calcium hydroxide lime--a new carbon dioxide absorbent: a rationale for judicious use of different absorbents. PMID- 11050517 TI - Comparison of ventilatory and haemodynamic effects of BIPAP and S-IMV/PSV for postoperative short-term ventilation in patients after coronary artery bypass grafting. AB - The aim of the present multiple cross-over study was to compare the effects of biphasic positive airway pressure (BIPAP) ventilation with synchronized intermittent mandatory ventilation combined with pressure support ventilation (S IMV/PSV) in sedated and awake patients after coronary artery bypass grafting (CABG) surgery. Twenty-four patients with no evidence of preoperative respiratory dysfunction and an uncomplicated intraoperative course were investigated. The patients were randomly assigned to one of two groups starting with either BIPAP or S-IMV/PSV mode. Haemodynamic measurements and blood gas analyses were performed during sedation with 2.0 mg kg(-1) h(-1) propofol in the primary mode, after switching to the alternative ventilatory mode, and in the primary mode again. The same sequence of measurements was repeated in awake patients who had reached extubation criteria. In awake patients, PSV was performed instead of S IMV. Statistical analysis of data was performed using non-parametric tests. Inspiratory peak pressure increased significantly during S-IMV/PSV in sedated patients in both groups. Other ventilatory parameters did not differ significantly between BIPAP and S-IMV/PSV in both groups. Similarly, haemodynamic parameters and blood-gas analyses did not vary with the ventilatory mode. Our results demonstrate that BIPAP ventilation has comparable effects on haemodynamics and pulmonary gas exchange compared with S-IMV/PSV and PSV when used for short-term ventilatory support in patients after cardiac surgery. PMID- 11050518 TI - Efficacy of prophylactic droperidol, ondansetron or both in the prevention of postoperative nausea and vomiting in major gynaecological surgery. A prospective, randomized, double-blind clinical trial. AB - We conducted a prospective, randomized, double-blind clinical trial comparing droperidol 1.25 mg intravenously (i.v.) (group 1, n = 30), ondansetron 4 mg i.v. (group 2, n = 30), or both (group 3, n = 30) in the prevention of postoperative nausea and vomiting (PONV) in the first 24 h following major gynaecological procedures under combined general and epidural anaesthesia. PONV was analysed by a linear nausea/vomiting score, incidence of nausea and vomiting, and the need for antiemetic rescue. Our results showed a similar incidence of nausea and vomiting in all groups (G1 33%, G2 40%, G3 43%). However, when comparisons were made according to the time of assessment, combination therapy resulted in significantly lower PONV than droperidol in the first hour (0% vs. 13%, P < 0.05) and second hour (0% vs. 13%, P < 0.05), and than ondansetron on the first hour (0% vs. 13%, P < 0.05). A trend persisted up to the fourth hour but was not statistically significant in either group. In conclusion, droperidol and ondansetron are effective agents in the prevention of PONV, and their combination seems to provide slightly better results than either drug alone. PMID- 11050520 TI - Accuracy of end-tidal carbon dioxide monitoring using the NBP-75 microstream capnometer. A study in intubated ventilated and spontaneously breathing nonintubated patients. AB - Arterial carbon dioxide partial pressure measurements using the NBP-75 microstream capnometer were compared with direct PaCO2 values in patients who were (a) not intubated and spontaneously breathing, and (b) patients receiving intermittent positive pressure ventilation of the lungs and endotracheal anaesthesia. Twenty ASA physical status I-III patients, undergoing general anaesthesia for orthopaedic or vascular surgery were included in a prospective crossover study. After a 20-min equilibration period following the induction of general anaesthesia, arterial blood was drawn from an indwelling radial catheter, while the end-tidal carbon dioxide partial pressure was measured at the angle between the tracheal tube and the ventilation circuit using a microstream capnometer (NBP-75, Nellcor Puritan Bennett, Plesanton, CA, USA) with an aspiration flow rate of 30 mL min(-1). Patients were extubated at the end of surgery and transferred to the postanaesthesia care unit, where end-tidal carbon dioxide was sampled through a nasal cannula (Nasal FilterLine, Nellcor, Plesanton, CA, USA) and measured using the same microstream capnometer. In each patient six measurements were performed, three during mechanical ventilation and three during spontaneous breathing. A good correlation between arterial and end tidal carbon dioxide partial pressure was observed both during mechanical ventilation (r = 0.59; P = 0.0005) and spontaneous breathing (r = 0.41; P = 0.001); while no differences in the arterial to end-tidal carbon dioxide tension difference were observed when patients were intubated and mechanically ventilated (7. 3 +/- 4 mmHg; CI95: 6.3-8.4) compared to values measured during spontaneous breathing in the postanesthesia care unit, after patients had been awakened and extubated (6.5 +/- 4.8 mmHg; CI95: 5. 2-7.8) (P = 0.311). The mean difference between the arterial to end-tidal carbon dioxide tension gradient measured in intubated and non-intubated spontaneously breathing patients was 1 +/- 6 mmHg (CI95: -11-+13). We conclude that measuring the end-tidal carbon dioxide partial pressure through a nasal cannula using the NBP-75 microstream capnometer provides an estimation of arterial carbon dioxide partial pressure similar to that provided when the same patients are intubated and mechanically ventilated. PMID- 11050519 TI - Itching after intrathecal morphine. Incidence and treatment. AB - This study was designed to determine whether low doses of intrathecal morphine still result in itching and it evaluates the outcome of a standardized treatment using promethazine and - for intractable itch - naloxone. Patients (n = 143) scheduled for total hip surgery were allocated to four groups (in a double blind manner) with bupivacaine 20 mg in 4 mL but different doses of intrathecal morphine: Group I, 0.025 mg, Group II, 0.05 mg, Group III, 0.1 mg and Group IV, 0.2 mg. The presence or absence of itching was noted every three hours for a 24-h period. When required, standardized treatment was provided. The incidence of itching was: Group I: 14. 3%; Group II: 21.6%; Group III: 48.6%; and, Group IV: 61.7%. Itch was treated by administering promethazine intramuscularly in 2.9% (Group I); 8.1% (Group II); 10.8% (Group III), and 8.9% (Group IV), respectively. Only in group IV there was a single patient who needed naloxone to treat itching. The incidence and severity of itching is a dose-related side-effect in the dose range of 0.025-0.2 mg of intrathecal morphine. Itching still occurs after the low doses of intrathecal morphine, but symptoms vanish after promethazine 25 mg intramuscularly. PMID- 11050521 TI - A comparison of the effects on postoperative pain relief of epidural analgesia started before or after surgery. AB - In a randomized, prospective clinical study pain relief and pulmonary function were compared after upper abdominal surgery when thoracic epidural analgesia was instituted either before or after surgery. Twenty-six patients admitted for surgery to treat gastro-oesophageal reflux received thoracic epidural analgesia as an adjunct to general anaesthesia either before or after surgery. Twelve patients received epidural mepivacaine 20 mg mL(-1) and morphine perioperatively. Another 14 patients received an epidural bolus of bupivacaine 2.5 mg mL(-1) and morphine after skin closure. Bupivacaine 2.5 mg mL(-1) with morphine was adminstered to all patients for three postoperative days. No intergroup differences were found regarding pain at rest and mobilization. The requirement for additional analgesics was similar in both groups as well as peak expiratory flow. Thoracic epidural analgesia that had already been induced before surgery, and was continued into the postoperative period, does not seem to add any advantage regarding pain relief and lung function compared with thoracic epidural analgesia instituted in the immediate postoperative period. PMID- 11050522 TI - Does etomidate increase postoperative nausea? A double-blind controlled comparison of etomidate in lipid emulsion with propofol for balanced anaesthesia. AB - In a double-blind randomized study, the incidence and severity of postoperative nausea and vomiting was investigated with a new formulation of etomidate (Etomidate-(R)Lipuro, B. Braun Melsungen AG, Germany) compared with propofol for induction of a balanced anaesthesia with isoflurane/fentanyl in air. The incidence and intensity of nausea was examined by use of a visual analogue scale (VAS; 0-100 mm) at 1, 2, between 6 and 8, and 24 h postoperatively. One-hundred and-sixty-four patients undergoing orthopedic procedures were studied. For etomidate vs. propofol, 14.6% vs. 14.2% male and 26.8% vs. 27.5% female patients were nauseated during the first two postoperative hours. The median rating for nausea remained below 5 mm at any time in both groups, i.e. the intensity of nausea was very low. The incidence of vomiting was higher in women receiving etomidate (26.8% vs. 10%). We conclude that etomidate does not increase nausea during the early postoperative period. PMID- 11050523 TI - Airway obstruction during general anaesthesia in a child with congenital tracheomalacia. AB - Fibreoptic bronchoscopy is often used to diagnose tracheomalacia under local anaesthesia. However, in children, general anaesthesia may be required due to difficulty in obtaining co-operation. A 1-yr-old girl with a suspected congenital tracheomalacia was scheduled for diagnostic fibreoptic bronchoscopy. During induction of anaesthesia by inhalation of increasing concentration of sevoflurane, spontaneous breathing became irregular and a partial airway obstruction occurred. Because vecuronium relieved the airway obstruction, the airway was managed using a laryngeal mask. No further airway obstruction occurred during fibrescopy under controlled ventilation, but when spontaneous breathing resumed, marked airway obstruction occurred. The trachea was intubated immediately. Caution is required to manage the airway without tracheal intubation during general anaesthesia in the patient with tracheomalacia. PMID- 11050524 TI - Anaesthesia for Proteus syndrome. AB - A 4-month-old boy with Proteus syndrome underwent a successful operation for a left abdominal mass due to hydroureter and hydronephrosis with left ureterovesical stenosis. The operation lasted 4.5 h under general anaesthesia; there were no anaesthetic complications. There is only one previous report on anaesthesia in a patient with Proteus syndrome. PMID- 11050525 TI - Who should provide anaesthesia? PMID- 11050526 TI - Knowledge about pulse oximetry among residents and nurses. PMID- 11050527 TI - Gastroscopy in awake and anaesthetized patients using a modified laryngeal mask. PMID- 11050528 TI - Extracellular ATP couples to cAMP generation and granulocytic differentiation in human NB4 promyelocytic leukaemia cells. AB - Priming of NB4 promyelocytic cells with all-trans retinoic acid, followed by extracellular ATP in the presence of a phosphodiesterase inhibitor, elevated cAMP and activated protein kinase A. The order of potency for cAMP production was ATP (EC50 = 95 +/- 13 micromol/L) > ADP > AMP = adenosine. The order of potency of ATP analogues was 2'- and 3'-O-(4-benzoylbenzoyl)-ATP (EC50 = 54 +/- 15 micromol/L) = adenosine 5'-O-(3-thio) triphosphate (EC50 = 66 +/- 4 micromol/L) > ATP > beta,gamma-methylene ATP (EC50 = 200 +/- 55 micromol/L). Adenosine 5'-O thiomonophosphate and adenosine 5'-O-(2-thio) diphosphate inhibited ATP-induced cAMP production. Differentiation also occurred as measured by increased expression of CD11b and N-formyl peptide receptor and changes in cell morphology. UTP did not elevate cAMP or induce differentiation, indicating that P2Y2, P2Y4, and P2Y6 receptors were not involved. The P2Y11 receptor, a cAMP-linked receptor on promyelocytic HL-60 cells, was detected in NB4 cells by reverse transcription polymerase chain reaction and northern blotting. This receptor has the same order of potency with respect to cAMP production as that observed in HL-60 cells. PMID- 11050529 TI - Binding of Tamm-Horsfall protein to complement 1q measured by ELISA and resonant mirror biosensor techniques under various ionic-strength conditions. AB - The purpose of the present study was to quantify the binding affinity between Tamm-Horsfall protein (THP) and complement 1q (C1q) using ELISA and a resonant mirror biosensor. In ELISA, immobilized THP was incubated with soluble C1q under both low and physiological ionic-strength conditions. Tamm-Horsfall protein bound C1q with an equilibrium dissociation constant (KD) of 1.9 +/- 0.6 nmol/L in low ionic-strength Tris buffers (20 mmol/L NaCl, pH 7.5) and with a lower affinity (KD of 13.4 +/- 4.7 nmol/L) in physiological-strength Tris buffers (154 mmol/L NaCl, pH 7.5). A resonant mirror biosensor, which monitors binding events in real time, was used to quantify the KD of this reaction, as well as to estimate the kinetic parameters. In these studies, THP and C1q bound with an association rate constant, kass, of 1.25 x 105 L/mol per s and a dissociation rate constant, kdiss, of 0.002-0.005/s. The calculated KD for the THP/C1q binding in low ionic strength buffers was higher (averages of 10-15 nmol/L) than that obtained by the ELISA, while physiological ionic-strength buffers still reduced the affinity of this binding by an order of magnitude. In conclusion, THP consistently bound C1q with high affinity using several techniques. At least a portion of this interaction involved electrostatic events, as demonstrated by the influence of ionic strength on the binding affinity. PMID- 11050530 TI - Special feature for the Olympics: effects of exercise on the immune system. Overview: exercise immunology. AB - The review articles in this special feature reflect the current status of knowledge in the field of exercise immunology, with a focus on how exercise affects the human immune system and the health implications for resistance to infections and neoplastic diseases. In an Olympic year, the emphasis of exercise immunology research tends to be on elite athletes and the prevention of infections in the quest for optimum performance. However, research presented in this issue also covers recreational athletes, as well as highlighting the benefits of exercise for our ageing population. PMID- 11050531 TI - Special feature for the Olympics: effects of exercise on the immune system: overview of the epidemiology of exercise immunology. AB - The application of epidemiological methods to exercise immunology is reviewed briefly, with particular reference to the possible influences of physical activity, exercise and training on susceptibility to upper respiratory infections. Available reports are arbitrarily rated in terms of limiting factors: the quality of the assessment of physical activity, the precision of diagnosis of upper respiratory infection and overall methodology. The pattern of physical activity has often been clearly established but, in part because of the problems associated with the competitive environment, assessments of infection and overall methodology have often been less than optimal. Although there is some evidence that susceptibility to infection is increased by either a single bout of very heavy activity or a period of heavy training, reports are far from unanimous, and in certain respects fail to meet the classical epidemiological criteria of a causal relationship. The issue is important to both the health and the success of the international competitor, and merits definitive investigation, using optimal methods to assess both activity patterns and infection. PMID- 11050532 TI - Special feature for the Olympics: effects of exercise on the immune system: exercise effects on systemic immunity. AB - Heavy exertion has acute and chronic influences on systemic immunity. In the resting state, the immune systems of athletes and non-athletes are more similar than disparate with the exception of NK cell activity, which tends to be elevated in athletes. Many components of the immune system exhibit adverse change after prolonged, heavy exertion. These immune changes occur in several compartments of the immune system and body (e.g. the skin, upper respiratory tract mucosal tissue, lung, blood and muscle). Although still open to interpretation, most exercise immunologists believe that during this 'open window' of impaired immunity (which may last between 3 and 72 h, depending on the immune measure) viruses and bacteria may gain a foothold, increasing the risk of subclinical and clinical infection. The infection risk may be amplified when other factors related to immune function are present, including exposure to novel pathogens during travel, lack of sleep, severe mental stress, malnutrition or weight loss. PMID- 11050533 TI - Special feature for the Olympics: effects of exercise on the immune system: overtraining effects on immunity and performance in athletes. AB - Overtraining is a process of excessive exercise training in high-performance athletes that may lead to overtraining syndrome. Overtraining syndrome is a neuroendocrine disorder characterized by poor performance in competition, inability to maintain training loads, persistent fatigue, reduced catecholamine excretion, frequent illness, disturbed sleep and alterations in mood state. Although high-performance athletes are generally not clinically immune deficient, there is evidence that several immune parameters are suppressed during prolonged periods of intense exercise training. These include decreases in neutrophil function, serum and salivary immunoglobulin concentrations and natural killer cell number and possibly cytotoxic activity in peripheral blood. Moreover, the incidence of symptoms of upper respiratory tract infection increases during periods of endurance training. However, all of these changes appear to result from prolonged periods of intense exercise training, rather than from the effects of overtraining syndrome itself. At present, there is no single objective marker to identify overtraining syndrome. It is best identified by a combination of markers, such as decreases in urinary norepinephrine output, maximal heart rate and blood lactate levels, impaired sport performance and work output at 110% of individual anaerobic threshold, and daily self-analysis by the athlete (e.g. high fatigue and stress ratings). The mechanisms underlying overtraining syndrome have not been clearly identified, but are likely to involve autonomic dysfunction and possibly increased cytokine production resulting from the physical stress of intense daily training with inadequate recovery. PMID- 11050534 TI - Special feature for the Olympics: effects of exercise on the immune system: infections and exercise in high-performance athletes. AB - The elite athlete has a potentially increased sensitivity to respiratory infections, rendering protective measures particularly important. Some other infections that may appear in clusters in the sports setting, such as gastroenteritis, leptospirosis, herpes simplex and viral hepatitis, also require special precautionary attention. Strenuous exercise during ongoing infection and fever may be hazardous and should always be avoided. In addition, early symptoms of infection warrant caution until the nature and severity of the infection become apparent. Because myocarditis may or may not be accompanied by fever, malaise or catarrhal symptoms, athletes should be informed about the symptoms suggestive of this disease. Although sudden unexpected death resulting from myocarditis is rare, exercise should be avoided whenever myocarditis is suspected. Guidelines are suggested for the management and counselling of athletes suffering from infections, including recommendations on when to resume training. Acute febrile infections are associated with decreased performance resulting from muscle wasting, circulatory deregulation and impaired motor coordination, which require variable amounts of time to become normalized once the infection is over. PMID- 11050535 TI - Special feature for the Olympics: effects of exercise on the immune system: effects of exercise on the immune system in the elderly population. AB - Immunosenescence is characterized by impaired cellular immune function concomitant with increased inflammatory activity. Immune dysfunction is associated with increased mortality risk in elderly people. An important part of human ageing is characterized by a decline in the ability of individuals to adapt to environmental stress. Exercise has been suggested as a prototype for studying the effects of stress factors on the cellular immune system. Studies of interactions between an acute bout of exercise and immune function may be a useful and an ethically acceptable tool to investigate cell trafficking, immune mobilization/deficiency and the acute phase response during physical stress situations in relation to human ageing. Elderly humans have a preserved ability to recruit T lymphocytes and NK cells in response to an acute bout of exercise. Physical exercise training programs do not result in major restoration of the senescent immune system in humans. However, highly conditioned elderly humans seem to have a relatively better preserved immune system, although it is not possible to conclude if this is linked to training or other lifestyle-related factors. PMID- 11050536 TI - Special feature for the Olympics: effects of exercise on the immune system: exercise and cytokines. AB - Strenuous exercise induces increased levels in a number of pro-inflammatory and anti-inflammatory cytokines, naturally occurring cytokine inhibitors and chemokines. Thus, increased plasma levels of TNF-alpha, IL-1, IL-6, IL-1 receptor antagonist, TNF receptors, IL-10, IL-8 and macrophage inflammatory protein-1 are found after strenuous exercise. The concentration of IL-6 increases up to 100 fold after a marathon race. The increase in IL-6 is tightly related to the duration of the exercise and there appears to be a logarithmic relationship. Furthermore, the increase in IL-6 is related to the intensity of exercise. Given the facts that IL-6, more than any other cytokine, is produced in large amounts in response to exercise, that IL-6 is produced locally in the skeletal muscle in response to exercise and that IL-6 is known to have growth factor abilities, it is likely that IL-6 plays a beneficial role and may be involved in mediating exercise-related metabolic changes. PMID- 11050537 TI - Special feature for the Olympics: effects of exercise on the immune system: exercise effects on mucosal immunity. AB - The present review examines the effects of exercise on mucosal immunity in recreational and elite athletes and the role of mucosal immunity in respiratory illness. Habitual exercise at an intense level can cause suppression of mucosal immune parameters, while moderate exercise may have positive effects. Saliva is the most commonly used secretion for measurement of secretory antibodies in the assessment of mucosal immune status. Salivary IgA and IgM concentrations decline immediately after a bout of intense exercise, but usually recover within 24 h. Training at an intense level over many years can result in a chronic suppression of salivary immunoglobulin levels. The degree of immune suppression and the recovery rates after exercise are associated with the intensity of exercise and the duration or volume of the training. Low levels of salivary IgM and IgA, particularly the IgA1 subclass, are associated with an increased risk of respiratory illness in athletes. Monitoring mucosal immune parameters during critical periods of training provides an assessment of the upper respiratory tract illness risk status of an individual athlete. The mechanisms underlying the mucosal immune suppression are unknown. PMID- 11050538 TI - Special feature for the Olympics: effects of exercise on the immune system: exercise-induced modulation of macrophage function. AB - Macrophages are important effector cells involved in phagocytosis, microbial killing and antitumour activity. Macrophages also display accessory cell function, in that they can present antigen to foster the development of T lymphocyte-mediated immunity. Recent work, including studies from this group, has demonstrated that acute and chronic exercise can affect many facets of macrophage biology. Manifestation of these effects depends on exercise intensity and duration, the function measured, the timing of measurement in relation to exercise and the concentration of the macrophage-activating stimulus. Exercise has potent stimulatory effects on phagocytosis, antitumour activity, reactive oxygen and nitrogen metabolism, and chemotaxis. Indeed, it has been shown that exercise training can increase macrophage antitumour activity in mice of different ages. However, not all functions are enhanced by exercise. Exercise induced reductions in macrophage MHC II expression and antigen-presentation capacity have been documented. These findings bring up the possibility that exercise, and perhaps other stressors, activate macrophages for effector functions while downregulating accessory cell functions. To a large extent, the mechanisms responsible for the exercise-induced changes in macrophage function remain unknown, but may depend on exercise-induced changes in neuroendocrine factors. Future studies need to explore the effects in a mechanistic way and provide documentation as to their physiological significance. PMID- 11050539 TI - Special feature for the Olympics: effects of exercise on the immune system: modification of immune responses to exercise by carbohydrate, glutamine and anti oxidant supplements. AB - Immunosuppression in athletes involved in heavy training is undoubtedly multifactorial in origin. Training and competitive surroundings may increase the athlete's exposure to pathogens and provide optimal conditions for pathogen transmission. Heavy prolonged exertion is associated with numerous hormonal and biochemical changes, many of which potentially have detrimental effects on immune function. Furthermore, improper nutrition can compound the negative influence of heavy exertion on immunocompetence. An athlete exercising in a carbohydrate depleted state experiences larger increases in circulating stress hormones and a greater perturbation of several immune function indices. The poor nutritional status of some athletes may predispose them to immunosuppression. For example, dietary deficiencies of protein and specific micronutrients have long been associated with immune dysfunction. Although it is impossible to counter the effects of all of the factors that contribute to exercise-induced immunosuppression, it has been shown to be possible to minimize the effects of many factors. Athletes can help themselves by eating a well-balanced diet that includes adequate protein and carbohydrate, sufficient to meet their energy requirements. This will ensure a more than adequate intake of trace elements without the need for special supplements. Consuming carbohydrate (but not glutamine or other amino acids) during exercise attenuates rises in stress hormones, such as cortisol, and appears to limit the degree of exercise-induced immunosuppression, at least for non-fatiguing bouts of exercise. Evidence that high doses of anti-oxidant vitamins can prevent exercise-induced immunosuppression is also lacking. PMID- 11050540 TI - Special feature for the Olympics: effects of exercise on the immune system: neuropeptides and their interaction with exercise and immune function. AB - It is known today that the immune system is influenced by various types of psychological and physiological stressors, including physical activity. It is well known that physical activity can influence neuropeptide levels both in the central nervous system as well as in peripheral blood. The reported changes of immune function in response to exercise have been suggested to be partly regulated by the activation of different neuropeptides and the identification of receptors for neuropeptides and steroid hormones on cells of the immune system has created a new dimension in this endocrine-immune interaction. It has also been shown that immune cells are capable of producing neuropeptides, creating a bidirectional link between the nervous and immune systems. The most common neuropeptides mentioned in this context are the endogenous opioids. The activation of endogenous opioid peptides in response to physical exercise is well known in the literature, as well as the immunomodulation mediated by opioid peptides. The role of endogenous opioids in the exercise-induced modulation of immune function is less clear. The present paper will also discuss the role of other neuroendocrine factors, such as substance P, neuropeptide Y and vasoactive intestinal peptide, and pituitary hormones, including growth hormone, prolactin and adrenocorticotrophin, in exercise and their possible effects on immune function. PMID- 11050541 TI - The molecular population genetics of regulatory genes. AB - Regulatory loci, which may encode both trans acting proteins as well as cis acting promoter regions, are crucial components of an organism's genetic architecture. Although evolution of these regulatory loci is believed to underlie the evolution of numerous adaptive traits, there is little information on natural variation of these genes. Recent molecular population genetic studies, however, have provided insights into the extent of natural variation at regulatory genes, the evolutionary forces that shape them and the phenotypic effects of molecular regulatory variants. These recent analyses suggest that it may be possible to study the molecular evolutionary ecology of regulatory diversification by examining both the extent and patterning of regulatory gene diversity, the phenotypic effects of molecular variation at these loci and their ecological consequences. PMID- 11050542 TI - Genetic structure of avian populations--allozymes revisited. AB - Selection on allozymes has sometimes been advanced as one explanation for the low levels of population differentiation detected in avian populations by the use of enzymatic markers. Comparisons of the amount of population subdivision (estimated by FST values or analogous indices) measured by enzymatic and mitochondrial DNA (mtDNA) markers in birds were seen as evidence for this because mtDNA typically produces a more structured picture of population subdivisions. In fact, when taking into account the smaller effective population size of mtDNA, nuclear and mitochondrial markers give concordant results. Some discrepancies still exist, but I suggest that some might originate from different amounts of nuclear vs. mitochondrial gene flow due to partial reproductive isolation. Variable number of tandem repeat (VNTR) loci do not provide a dramatically different picture of population structures in birds compared to allozymes. Although more tests are needed, such as comparing the amount of genetic structure detected in the same populations with allozymes and microsatellites, the low levels of population subdivision measured with allozymes in birds seem to reflect historical and demographic processes and would not appear to result from any peculiarities of bird enzymatic loci. PMID- 11050543 TI - Multiple gene genealogies reveal recent dispersion and hybridization in the human pathogenic fungus Cryptococcus neoformans. AB - Cryptococcus neoformans (= Filobasidiella neoformans) is a significant emerging fungal pathogen of humans. To understand the evolution of this pathogen, 34 strains were obtained from various locations around the world and fragments of four genes were sequenced from each. These strains represented all three varieties and five serotypes. The four sequenced genes are: (i) the mitochondrial large ribosomal subunit RNA; (ii) the internal transcribed spacer region of the nuclear rRNA, including ITS1, 5.8S rRNA subunit and ITS2; (iii) orotidine monophosphate pyrophosphorylase; and (iv) diphenol oxidase. Phylogenetic analyses indicated considerable divergence among lineages, which corresponded to the current classification of C. neoformans into three varieties. However, there is no apparent phylogeographic pattern. Significant incongruences were observed among gene genealogies. The analyses indicated that the major lineages in C. neoformans diverged tens of millions of years ago but have undergone recent dispersion and hybridization. PMID- 11050544 TI - Mitochondrial DNA phylogeography of Acrossocheilus paradoxus (Cyprinidae) in Taiwan. AB - Nucleotide sequences of 3' end of the cytochrome b gene, tRNA genes, D-loop control region, and the 5' end of the 12S rRNA of mitochondrial DNA (mtDNA) were used to assess the genetic and phylogeographic structure of Acrossocheilus paradoxus populations, a Cyprinidae fish of Taiwan. A hierarchical examination of populations in 12 major streams from three geographical regions using an analysis of molecular variance (AMOVA) indicates high genetic differentiation both among populations (PhiST = 0.511, P < 0.001) and among regions (PhiCT = 0.368, P < 0.001). Limited migration largely contributed to the genetic differentiation. High nucleotide diversity (1.13%) and haplotype diversity (0.80%) were detected among populations. The degree of genetic differentiation was correlated with geographical distance between populations, a result consistent with the one dimensional stepping stone models. A neighbour-joining tree recovered by (DAMBE) supports the pattern of isolation by distance and reveals a closer relationship between populations of the central and southern regions. A minimum spanning network based on nucleotide substitutions reflected migration routes from populations of the central region to the northern and southern regions, respectively. Postglacial colonization and expansion can explain the phylogeographical pattern. Single and ancient migration events may have allowed the northern region to attain the monophyly of mtDNA alleles. In contrast, most populations within geographical regions are either paraphyletic or polyphyletic due to the relatively shorter time period for coalescence. Both low haplotype number and genetic variability suggest a bottleneck event in the Chingmei population of northern Taiwan. Based on coalescence theory, the monophyly of the Tungkang population of the southern region may be associated with a founder event. PMID- 11050545 TI - Chloroplast DNA polymorphism reveals little geographical structure in Castanea sativa Mill. (Fagaceae) throughout southern European countries. AB - The distribution of haplotypic diversity of 38 European chestnut (Castanea sativa Mill.) populations was investigated by PCR/RFLP analysis of regions of the chloroplast and mitochondrial genomes in order to shed light on the history of this heavily managed species. The rapid expansion of chestnut starting from 3000 years ago is strongly related to human activities such as agricultural practice. This demonstrates the importance of human impact, which lasted some thousands of years, on the present-day distribution of the species. No polymorphism was detected for the single mitochondrial analysed region, while a total of 11 different chloroplast (cp) haplotypes were scored. The distribution of the cpDNA haplotypes revealed low geographical structure of the genetic diversity. The value of population subdivision, as measured by GSTc, is strikingly lower than in the other species of the family Fagaceae investigated. The actual distribution of haplotypic diversity may be explained by the strong human impact on this species, particularly during the Roman civilization of the continent, and to the long period of cultivation experienced during the last thousand years. PMID- 11050546 TI - Comparative avian phylogeography of Cameroon and equatorial Guinea mountains: implications for conservation. AB - We illustrate the use of Faith's 'Phylogenetic Diversity' measure to compare the phylogeographic structure of two bird species with patterns of avian endemism across six mountains in Cameroon and Equatorial Guinea. The Mountain Greenbul and Cameroon Blue-headed Sunbird showed phylogeographic patterns that together defined three biogeographic regions: Bioko, Mt. Cameroon, and the northern mountains of Cameroon. In contrast, the distributions of endemic species were largely a function of geographical distance, with close mountains sharing more endemic species than distant mountains. Moreover, for both species, populations on Mt. Cameroon were distinctive with respect to the ecologically relevant character bill size. Our results, while preliminary, illustrate the utility of a comparative approach for identifying geographical regions that harbour evolutionarily distinct populations and caution against using only the distributional patterns of endemics to prioritize regions for conservation. Results show that patterns of endemism may not be concordant with patterns of phylogenetic diversity nor morphological variation in a character important in fitness. While incorporation of additional species from unrelated taxa will be necessary to draw definitive conclusions about evolutionarily distinct regions, our preliminary results suggest a conservation approach for the Afromontane region of the Gulf of Guinea that would: (i) emphasize protection of both Bioko and Mt. Cameroon, thereby maximizing preservation of within-species phylogenetic and morphologic diversity; (ii) emphasize protection within the northern mountains to further conserve intraspecific phylogenetic diversity and maximize protection of endemic species. PMID- 11050547 TI - Experimental evaluation of the usefulness of microsatellite DNA for detecting demographic bottlenecks. AB - Evolutionary and conservation biologists often use molecular markers to evaluate whether populations have experienced demographic bottlenecks that resulted in a loss of genetic variation. We evaluated the utility of microsatellites for detection of recent, severe bottlenecks and compared the amounts of genetic diversity lost in bottlenecks of different sizes. In experimental mesocosms, we established replicate populations by releasing 1, 2, 4 or 8 pairs of the western mosquitofish, Gambusia affinis (Poeciliidae). Using eight polymorphic microsatellite loci, we quantified seven indices of genetic diversity or change that have been used to assess the effects of demographic bottlenecks on populations. We compared indices for the experimentally bottlenecked populations to those for the source population and examined differences between populations established with different numbers of founders. Direct count heterozygosity and the proportion of polymorphic loci were not very sensitive to genetic changes that resulted from the experimental bottlenecks. Heterozygosity excess and expected heterozygosity were useful to varying degrees in the detection of bottlenecks. Allelic diversity and temporal variance in allele frequencies were most sensitive to genetic changes that resulted from the bottlenecks, and the temporal variance method was slightly more correlated with bottleneck size than was allelic diversity. Based on comparisons to a previous study with allozymes, heterozygosity, temporal variance in allele frequencies and allelic diversity, but not proportion of polymorphic loci, appear to be more sensitive to demographic bottlenecks when quantified using microsatellites. We found that analysis of eight highly polymorphic loci was sufficient to detect a recent demographic bottleneck and to obtain an estimate of the magnitude of bottleneck severity. PMID- 11050548 TI - Increase of genetic variation over time in a recently founded population of great reed warblers (Acrocephalus arundinaceus) revealed by microsatellites and DNA fingerprinting. AB - Genetic similarity within pairs of individuals was examined using both 10 polymorphic microsatellite loci and multi-locus DNA fingerprinting profiles in a semi-isolated population of great reed warblers at Lake Kvismaren, south Central Sweden, in 1987-1993. The population was founded by a few individuals in 1978, followed by a gradual increase in numbers until 1988, since when the population has remained relatively stable with about 60 breeding birds. We have previously found that high genetic similarity between pair-mates in the population during the early part of the study period reduced egg hatching success, and hence reproductive success. The measures of pairwise genetic similarity, microsatellite allele sharing and DNA fingerprinting band sharing, were highly correlated with pedigree-based relatedness. Both microsatellite and DNA fingerprinting similarities between pair-mates declined significantly over the study period, and the pattern was most pronounced in the DNA fingerprinting data. Analyses restricted to the microsatellite data showed that the average annual microsatellite similarity between pairwise combinations of individuals, as well as individual homozygosity in males, declined significantly over the study period, and that several immigrants carrying novel alleles entered the population during the study. Hence, the temporal decline in genetic similarity of mates in the population is probably a consequence of increased immigration, facilitated by the recent expansion of the species in the region. These results suggest that the population has now recovered genetically, or is in the process of recovering, from a recent founder event. PMID- 11050549 TI - Genetic variability among endangered Chinese giant salamanders, Andrias davidianus. AB - The endangered Chinese giant salamander (Andrias davidianus) is endemic to mainland China. Genetic divergence among six populations of the species was investigated by means of isozyme electrophoresis and mitochondrial DNA (mtDNA) sequences. Forty allozyme loci were resolved for all populations; the amount of genetic divergence among populations was comparable to that in other amphibians. mtDNA sequences showed a similar level of divergence. The population from Huangshan is distinct from other populations, indicating the existence of localized divergence. Both allozyme and mtDNA data failed to associate the populations into a pattern corresponding to the three Chinese river systems, which may be the consequence of human relocation. Conservation policies should emphasize the protection of localized populations and cessation of human facilitated introductions. Future studies should focus on investigating the divergence among localized populations from isolated mountain regions, particularly using more fine-grained techniques such as microsatellite DNA. PMID- 11050550 TI - A universal molecular method for identifying underground plant parts to species. AB - As part of a large project to determine rooting depth and resource uptake on the Edwards Plateau of central Texas, we developed a DNA-based technique that allows the below-ground parts of all plants to be identified to the level of genus and usually to species. Identification is achieved by comparing DNA sequences of the internal transcribed spacer (ITS) region of the 18S-26S nuclear ribosomal DNA repeat, derived from below-ground plant material, with a reference ITS region database for plants at a site. The method works throughout plants because the plant ITS region can be PCR amplified using a set of universal primers. Congeneric species can usually be identified because the ITS region evolves relatively rapidly. In our study, all roots were easily identified to the level of genus; most congeneric species were identified solely by ITS sequence differences but some required a combination of ITS sequence data and above-ground surveys of species at a site. In addition to showing the feasibility and efficacy of our technique, we compare it with another DNA-based technique used to identify below-ground plant parts. Finally, we also describe a DNA extraction and purification technique that reliably provides high-quality DNA of sufficient quantity from roots so that PCR can be readily accomplished. Our technique should allow the below-ground parts of plants in any system to be identified and thereby open new possibilities for the study of below-ground plant communities. PMID- 11050551 TI - Microsatellite analysis of North American wapiti (Cervus elaphus) populations. AB - Eleven populations of wapiti (Cervus elaphus) were analysed for genetic diversity using 12 microsatellite loci. Samples were taken from Vancouver Island, British Columbia; Burwash and French River herds in Ontario; Ya Ha Tinda Ranch, Alberta; and Banff, Elk Island, Jasper, Kootenay, Riding Mountain, Yellowstone and Yoho National Parks. Overall, wapiti populations have on average three to four alleles per locus and an average expected heterozygosity that ranged from 25.75 to 52.85%. The greatest genetic distances were observed between the Vancouver population and all other populations. Using the assignment test, Roosevelt wapiti (C. e. roosevelti Merriam 1897) assigned only to the Vancouver Island population. The distance and assignment values suggest a divergence of the Roosevelt wapiti from other populations and support the subspecific status for the Vancouver Island population. No evidence was found for the existence of unique Eastern wapiti (C. e. canadensis Erxleben 1777) in the Burwash or French River herds in Ontario. The overlapping distribution of genotypes from indigenous populations from Riding Mountain, Elk Island and Yellowstone National Parks suggests that wapiti were once a continuous population before settlers decimated their numbers. The lack of differentiation between these populations raises questions about the status of Manitoban (C. e.manitobensis Millais 1915) and Rocky Mountain (C. e.nelsoni Bailey 1935) subspecies. PMID- 11050552 TI - Across the Southern Alps by river capture? Freshwater fish phylogeography in South Island, New Zealand. AB - We used DNA analysis of galaxiid fish to test a hypothesis of localized headwater capture in South Island, New Zealand. The restricted western, but widespread eastern, distributions of three nonmigratory freshwater fish species suggest that part of the east-flowing Waiau River has been captured by the west-flowing Buller River. However, mitochondrial control region (Kimura 2-parameter distance = 4.1 5.4%) and microsatellite flanking sequences do not support a relationship between Waiau (N = 4 fish sequences) and western populations (N = 8) of Galaxias vulgaris. Instead, the point of capture is probably to the north-east, perhaps the Nelson lakes region. Phylogenetic analysis indicates that western populations, along with populations in the north-east (N = 18), represent a previously unidentified monophyletic Evolutionarily Significant Unit, possibly a cryptic species. We suggest a general caveat for zoogeographic conclusions based on distributional data alone. PMID- 11050553 TI - Dietary separation of sympatric carnivores identified by molecular analysis of scats. AB - We studied the diets of four sympatric carnivores in the flooding savannas of western Venezuela by analysing predator DNA and prey remains in faeces. DNA was isolated and a portion of the cytochrome b gene of the mitochondrial genome amplified and sequenced from 20 of 34 scats. Species were diagnosed by comparing the resulting sequences to reference sequences generated from the blood of puma (Puma concolor), jaguar (Panthera onca), ocelot (Leopardus pardalus) and crab eating fox (Cerdocyon thous). Scat size has previously been used to identify predators, but DNA data show that puma and jaguar scats overlap in size, as do those of puma, ocelot and fox. Prey-content analysis suggests minimal prey partitioning between pumas and jaguars. In field testing this technique for large carnivores, two potential limitations emerged: locating intact faecal samples and recovering DNA sequences from samples obtained in the wet season. Nonetheless, this study illustrates the tremendous potential of DNA faecal studies. The presence of domestic dog (Canis familiaris) in one puma scat and of wild pig (Sus scrofa), set as bait, in one jaguar sample exemplifies the forensic possibilities of this noninvasive analysis. In addition to defining the dietary habits of similar size sympatric mammals, DNA identifications from faeces allow wildlife managers to detect the presence of endangered taxa and manage prey for their conservation. PMID- 11050554 TI - Fragmented forests, evolving flies: molecular variation in African populations of Drosophila teissieri. AB - Microsatellite variation from eight loci was studied in five populations of Drosophila teissieri, a fruit-fly found only in the rain forests of sub-Saharan Africa. Five noncontiguous rain forest sites (from Tanzania, Gabon and Ivory Coast) were sampled to measure the effects of historical forest fragmentation on population structure in an obligatory forest-dwelling species. The Ivory Coast and Gabon populations showed a wider range of alleles, different modal alleles and had a higher genetic diversity than the three East African populations. As could be expected, genetic differentiation (FST) was significantly correlated with physical distance, but the westernmost population (Ivory Coast) showed values that were intermediate between the central (Gabon) and Eastern (Tanzania) populations. A migration-drift equilibrium in a stable continuum of populations did not appear adequate to describe the observed distribution. It seems probable that the species has undergone abrupt changes involving isolation, merging and migration of populations, as a consequence of repeated waves of forest fragmentation and coalescence. PMID- 11050555 TI - Complex mutations in a high proportion of microsatellite loci from the protozoan parasite Plasmodium falciparum. AB - Microsatellite loci are generally assumed to evolve via a stepwise mutational process and a battery of statistical techniques has been developed in recent years based on this or related mutation models. It is therefore important to investigate the appropriateness of these models in a wide variety of taxa. We used two approaches to examine mutation patterns in the malaria parasite Plasmodium falciparum: (i) we examined sequence variation at 12 tri-nucleotide repeat loci; and (ii) we analysed patterns of repeat structure and heterozygosity at 114 loci using data from 12 laboratory parasite lines. The sequencing study revealed complex patterns of mutation in five of the 12 loci studied. Alleles at two loci contain indels of 24 bp and 57 bp in flanking regions, while in the other three loci, blocks of imperfect microsatellites appear to be duplicated or inserted; these loci essentially consist of minisatellite repeats, with each repeat unit containing four to eight microsatellites. The survey of heterozygosity revealed a positive relationship between repeat number and microsatellite variability for both di- and trinucleotides, indicating a higher mutation rate in loci with longer repeat arrays. Comparisons of levels of variation in different repeat types indicate that the mutation rate of dinucleotide-bearing loci is 1.6-2.1 times faster than trinucleotides, consistent with the lower mean number of repeats in trinucleotide-bearing loci. However, despite the evidence that microsatellite arrays themselves are evolving in a manner consistent with stepwise mutation model in P. falciparum, the high frequency of complex mutations precludes the use of analytical tools based on this mutation model for many microsatellite-bearing loci in this protozoan. The results call into question the generality of models based on stepwise mutation for analysing microsatellite data, but also demonstrate the ease with which loci that violate model assumptions can be detected using minimal sequencing effort. PMID- 11050556 TI - Genetic lineages and postglacial colonization of grayling (Thymallus thymallus, Salmonidae) in Europe, as revealed by mitochondrial DNA analyses. AB - In stark contrast to other species within the Salmonidae family, phylogeographic information on European grayling, Thymallus thymallus, is virtually nonexistent. In this paper, we utilized mitochondrial DNA polymerase chain reaction restriction fragment length polymorphism (mtDNA PCR-RFLP) and sequence variation to infer the postglacial dispersal routes of T. thymallus into and within northern Europe, and to locate geographically, potential evolutionarily distinct populations. Mitochondrial analyses revealed a total of 27 T. thymallus haplotypes which clustered into three distinct lineages. Average pairwise interlineage divergence was four and nine times higher than average intralineage divergence for RFLP and sequence data, respectively. Two European grayling individuals from the easternmost sample in Russia exhibited haplotypes more genetically diverged from any T. thymallus haplotype than T. arcticus haplotype, and suggested that hybridization/introgression zone of these two sister species may extend much further west than previously thought. Geographic division of the lineages was generally very clear with northern Europe comprising of two genetically differentiated areas: (i) Finland, Estonia and north-western Russia; and (ii) central Germany, Poland and western Fennoscandia. Average interpopulation divergence in North European T. thymallus was 10 times higher than that observed in a recent mtDNA study of North American T. arcticus. We conclude that (i) North European T. thymallus populations have survived dramatic Pleistocene temperature oscillations and originate from ancient eastern and central European refugia; (ii) genetic divergence of population groups within northern Europe is substantial and geographically distinct; and (iii) the remainder of Europe harbours additional differentiated assemblages that likely descend from a Danubian refugium. These findings should provide useful information for developing appropriate conservation strategies for European grayling and exemplify a case with a clear need for multinational co-operation for managing and conserving biodiversity. PMID- 11050557 TI - Genetic diversity in an endangered alpine plant, Eryngium alpinum L. (Apiaceae), inferred from amplified fragment length polymorphism markers. AB - Eryngium alpinum L. is an endangered species found across the European Alps. In order to obtain base-line data for the conservation of this species, we investigated levels of genetic diversity within and among 14 populations from the French Alps. We used the amplified fragment length polymorphism (AFLP) technique with three primer pairs and scored a total of 62 unambiguous, polymorphic markers in 327 individuals. Because AFLP markers are dominant, within-population genetic structure (e.g. FIS) could not be assessed. Analyses based either on the assumption of random-mating or on complete selfing lead to very similar results. Diversity levels within populations were relatively high (mean Nei's expected heterozygosity = 0.198; mean Shannon index = 0.283), and a positive correlation was detected between both genetic diversity measurements and population size (Spearman rank correlation: P = 0. 005 and P = 0.002, respectively). Moreover, FST values and exact tests of differentiation revealed high differentiation among populations (mean pairwise FST = 0.40), which appeared to be independent of geographical distance (nonsignificant Mantel test). Founder events during postglacial colonizations and/or bottlenecks are proposed to explain this high but random genetic differentiation. By contrast, we detected a pattern of isolation by distance within populations and valleys. Predominant local gene flow by pollen or seed is probably responsible for this pattern. Concerning the management of E. alpinum, the high genetic differentiation leads us to recommend the conservation of a maximum number of populations. This study demonstrates that AFLP markers enable a quick and reliable assessment of intraspecific genetic variability in conservation genetics. PMID- 11050558 TI - Ericoid mycorrhizal fungi are common root associates of a Mediterranean ectomycorrhizal plant (Quercus ilex). AB - Mycorrhiza samples of neighbouring Quercus ilex and Erica arborea plants collected in a postcutting habitat were processed to see whether plants differing in mycorrhizal status harbour the same root endophytes. Three experiments were performed in parallel: (i) isolation, identification and molecular characterization of fungi from surface-sterilized roots of both plant species; (ii) re-inoculation of fungal isolates on axenic E. arborea and Q. ilex seedlings; (iii) direct inoculation of field-collected Q. ilex ectomycorrhizas onto E. arborea seedlings. About 70 and 150 fungal isolates were obtained from roots of Q. ilex and E. arborea, respectively. Among them, Oidiodendron species and five cultural morphotypes of sterile isolates formed typical ericoid mycorrhizas on E. arborea in vitro. Fungi with such mycorrhizal ability were derived from both host plants. Isolates belonging to one of these morphotypes (sd9) also exhibited an unusual pattern of colonization, with an additional extracellular hyphal net. Ericoid mycorrhizas were also readily obtained by direct inoculation of E. arborea seedlings with Q. ilex ectomycorrhizal tips. Polymerase chain-restriction fragment length polymorphism and random amplified polymorphic DNA analyses of the shared sterile morphotypes demonstrate, in the case of sd9, the occurrence of the same genet on the two host plants. These results indicate that ericoid mycorrhizal fungi associate with ectomycorrhizal roots, and the ecological significance of this finding is discussed. PMID- 11050559 TI - Microsatellite analysis of population structure and genetic differentiation within and between populations of the root vole, Microtus oeconomus in the Netherlands. AB - Eight microsatellite markers for the root vole (Microtus oeconomus) were developed to assess the amount of genetic variation for nine Dutch root vole populations from four different regions, and to evaluate the degree of differentiation and isolation. All eight microsatellite loci were found to be highly variable with observed heterozygosity values ranging from 0.61 to 0.82. These values are similar to those observed for more distant populations from Norway, Finland and Germany. Therefore, the populations seem not particularly depauperate of genetic variation at the microsatellite level. Genetically, the Dutch populations were found to have diverged considerably. Pairwise comparisons of all populations studied revealed FST values significantly greater than zero for most comparisons. However, the magnitude of these values considerably depends on the compared population pair. The level of differentiation between local populations within Dutch regions is generally significantly lower than the differentiation between Dutch regions. The level of differentiation between Dutch regions, however, is not significantly different from that between populations of larger geographical distance. This implies that the regional Dutch populations are both isolated from each other and from other European populations. The observation that even local populations show low but significant genetic differentiation may be indicative for progressive isolation of these populations. PMID- 11050560 TI - TCS: a computer program to estimate gene genealogies. PMID- 11050561 TI - Polymorphic microsatellite DNA markers in the asiatic black bear Ursus thibetanus. PMID- 11050562 TI - Microsatellites in the hermaphroditic snail, Lymnaea truncatula, intermediate host of the liver fluke, Fasciola hepatica. PMID- 11050563 TI - Characterization and isolation of DNA microsatellite primers in wood mice (Apodemus sylvaticus, Rodentia). PMID- 11050564 TI - The isolation and characterization of microsatellites from Anopheles dirus mosquitoes. PMID- 11050565 TI - Microsatellites for studies of ecology, behaviour, and evolution in Yarrow's spiny lizard (Sceloporus jarrovii). PMID- 11050566 TI - Microsatellite markers for the deer mouse Peromyscus maniculatus. PMID- 11050567 TI - Isolation and characterization of microsatellites in the endemic species Centaurea corymbosa Pourret (Asteraceae) and other related species. PMID- 11050568 TI - Characterization of microsatellite loci from the mountain vizcacha Lagidium viscacia and their use for the plains vizcacha Lagostomus maximus. PMID- 11050569 TI - Identification of polymorphic autosomal and sex chromosome specific DNA microsatellites in the bushcricket, Poecilimon hoelzeli (Orthoptera, Tettigonioidea, Phaneropteridae). PMID- 11050570 TI - Characterization and isolation of DNA microsatellite primers in hyrax species (Procavia johnstoni and Heterohyrax brucei, hyracoidea). PMID- 11050571 TI - Isolation and characterization of microsatellite loci in the red-crowned crane Grus japonensis. PMID- 11050572 TI - Isolation and characterization of microsatellites in the moss species Polytrichum formosum. PMID- 11050573 TI - Highly polymorphic microsatellite loci from the Dominican anole (Anolis oculatus) and their amplification in other bimaculatus series anoles. PMID- 11050574 TI - Isolation and characterization of microsatellite loci in marsupial gliders (Petaurus norfolcensis, P. breviceps and P. gracilis). PMID- 11050575 TI - Development of microsatellite markers in the tropical tree Neobalanocarpus heimii (Dipterocarpaceae). PMID- 11050576 TI - Isolation and characterization of four polymorphic microsatellite loci in Forficula auricularia L. (Dermaptera: Forficulidae). PMID- 11050577 TI - Dermatologic diseases of the breast and nipple. AB - Breast and nipple skin is commonly affected by various inflammatory and neoplastic processes. Despite this fact, many physicians are unaware of the spectrum of diseases that can involve this area. Because breast and nipple skin represents a cosmetically, sexually, and functionally important entity to most patients, awareness of these disease entities is invaluable. This article reviews the normal anatomy of the breast, cutaneous manifestations of neoplastic processes that can present in these areas, and common inflammatory diseases of the breast and nipple skin. PMID- 11050578 TI - Merkel cell carcinoma: report of 10 cases and review of the literature. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a rare primary neuroendocrine skin tumor that usually arises in the head and neck or the extremities of elderly patients. Because of the limitation of retrospective data, optimal treatment is not well defined. OBJECTIVE: Our purpose was to present the clinical course and treatment of 10 patients with MCC and review the published literature on MCC. METHOD: We conducted a retrospective analysis and obtained detailed clinical information for all 10 patients treated for MCC at our institution from 1986 through 1998. The medical literature was also reviewed for natural history and treatment recommendations using MEDLINE search. RESULTS: Five men and 5 women received their treatment between 1986 and 1998 for MCC (5 had stage IA disease, 4 stage IB, 1 stage II). The mean age was 70.3 years (range, 47-86 years). Seven tumors were located on the head and neck and 3 on extremities. Five of 10 patients had a relapse (mean time before recurrence, 5.7 months) (range, 2 weeks 20 months); one patient had local recurrence, one had regional lymph node recurrence, and 3 had both local and regional lymph node recurrence. In 4 patients systemic metastases developed. Long survival is also noted (6 to > 164 months); 4 patients died of MCC. After initial surgery, 9 patients received radiotherapy at some point and 3 patients also received chemotherapy. Five of 10 patients had 13 previously treated or coexisting malignant neoplasms. In one patient MCC developed in a previously irradiated field. Review of 875 cases showed a male/female ratio of 1.5:1; location of tumors was as follows: head and neck, 47%; extremities, 40%; trunk, 8%; unknown primary site, 5%. Local recurrence was observed in 25%, regional lymph node involvement in 52%, distant metastasis in 34%, and MCC was a cause of death in 34%. CONCLUSION: MCC has a high incidence of locoregional recurrence with distant metastases that is more common with higher stage lesions. Early local management of smaller lesions results in good long-term survival. It is not known whether prophylactic lymph node dissection and/or radiation and adjuvant radiation increases survival. Long survival can be achieved after treating locoregional recurrence. The role of chemotherapy is still controversial and should be considered in patients with advanced disease and those not thought to be candidates for surgery. PMID- 11050579 TI - Lack of efficacy of finasteride in postmenopausal women with androgenetic alopecia. AB - BACKGROUND: Finasteride, an inhibitor of type 2 5alpha-reductase, decreases serum and scalp dihydrotestosterone (DHT) by inhibiting conversion of testosterone to DHT and has been shown to be effective in men with androgenetic alopecia (AGA). The effects of finasteride in women with AGA have not been evaluated. OBJECTIVE: The purpose of this study was to evaluate the efficacy of finasteride in postmenopausal women with AGA. METHODS: In this 1-year, double-blind, placebo controlled, randomized, multicenter trial, 137 postmenopausal women (41-60 years of age) with AGA received finasteride 1 mg/day or placebo. Efficacy was evaluated by scalp hair counts, patient and investigator assessments, assessment of global photographs by a blinded expert panel, and histologic analysis of scalp biopsy specimens. RESULTS: After 1 year of therapy, there was no significant difference in the change in hair count between the finasteride and placebo groups. Both treatment groups had significant decreases in hair count in the frontal/parietal (anterior/mid) scalp during the 1-year study period. Similarly, patient, investigator, and photographic assessments as well as scalp biopsy analysis did not demonstrate any improvement in slowing hair thinning, increasing hair growth, or improving the appearance of the hair in finasteride-treated subjects compared with the placebo group. Finasteride was generally well tolerated. CONCLUSION: In postmenopausal women with AGA, finasteride 1 mg/day taken for 12 months did not not increase hair growth or slow the progression of hair thinning. PMID- 11050581 TI - Hanifin's and Rajka's minor criteria for atopic dermatitis: which do 2-year-olds exhibit? AB - BACKGROUND: In 1980 Hanifin and Rajka published major and minor criteria for atopic dermatitis (AD). Despite the early age at onset of AD, there are few prospective studies in young children of the prevalence of signs and symptoms meeting the minor criteria. OBJECTIVE: Our purpose was to identify those of Hanifin's and Rajka's minor criteria that are most frequent in 2-year-old children with AD and in controls. METHODS: Consecutive patients with AD (n = 221), 24 months of age or younger, were followed up to 2 years, when they were re examined. The minor criteria were divided into 33 subcriteria, 29 of which were examined. Controls (n = 99), matched for age and sex, with no history of eczema at 2 years of age were examined in the same way. RESULTS: At the 2-year examination 157 of 221 had ongoing AD. Seven minor criteria were met in more than one fourth of these children, namely xerosis (100%), course influenced by environmental factors (87%), facial erythema (54%), skin reactions provoked by ingested food (39%), itch when sweating (34%), positive skin prick test (29%), and hand eczema (28%). In the control group, only xerosis (40%), facial erythema (25%), and skin reactions provoked by ingested food (9%) were present in 4% or more. CONCLUSION: Approximately half of the 29 criteria investigated were met in 3% or fewer of the cases, indicating that they may not be of much help to the clinician. Of the minor criteria of Hanifin and Rajka, only xerosis, course influenced by environmental factors, and facial erythema were seen in a majority of patients and would therefore be useful in the diagnosis of AD. PMID- 11050580 TI - Concomitant administration of vitamin E does not change the side effects of isotretinoin as used in acne vulgaris: a randomized trial. AB - BACKGROUND: Isotretinoin treatment is frequently associated with reversible, dose related side effects. Recent studies claimed that combining vitamin E with high dose isotretinoin ameliorated isotretinoin-induced side effects. OBJECTIVE: The purpose of this double-blind, randomized study was to determine the effects of a fixed dose of vitamin E on the side effects of isotretinoin for treatment resistant acne vulgaris. METHODS: One hundred forty subjects were randomly assigned to one of two treatment programs with isotretinoin (1 mg/kg) together with either vitamin E (800 IU/day) or a vitamin E placebo for 20 weeks. The incidence, severity, and duration of the side effects (eg, dry eyes, dry lips) were assessed. RESULTS: A fixed 800 IU/day dose of vitamin E did not improve the incidence, severity, or duration of side effects associated with isotretinoin (1 mg/kg per day). CONCLUSION: Vitamin E did not significantly ameliorate retinoid side effects when combined with 1 mg/kg of isotretinoin in the treatment of acne. PMID- 11050582 TI - Relative frequency of various forms of primary cutaneous lymphomas. AB - BACKGROUND: Data on the relative frequency of the various forms of primary cutaneous lymphomas (PCLs) are largely limited to European institutions. OBJECTIVE: Our purpose was to document the relative frequencies of various PCLs seen at 3 US institutions with active cutaneous lymphoma programs and to compare those with the European data. METHODS: Included in this study are newly registered patients seen at MCP Hahnemann University, New York University, and the University of California, San Francisco from July 1, 1995 to June 30, 1998. RESULTS: A total of 755 patients were seen. The frequency distribution of the major diagnostic groups was as follows: mycosis fungoides/Sezary syndrome, 82.3%; lymphomatoid papulosis, 12.6% (including patients with associated mycosis fungoides/Sezary syndrome); CD30(+) anaplastic large-cell lymphoma, 0.9%; peripheral T-cell lymphomas, 2.9%; B-cell lymphoma, 4.5%. CONCLUSION: The most striking finding is the much lower relative frequency of primary cutaneous B-cell lymphomas at US institutions (4.5%) versus the approximately 20% reported by European groups. The reason for this difference requires further study. PMID- 11050583 TI - Congenital fascial dystrophy: abnormal composition of the fascia. AB - BACKGROUND: A scleroderma-like genetic disease, congenital fascial dystrophy, probably a variant of stiff skin syndrome described by Esterly and McKusick, was found to be related to genetically determined fascial abnormalities. Our previous electronmicroscopic study disclosed as a main pathologic finding presence of giant amianthoid-like collagen fibrils in the affected fascia. OBJECTIVE: The aim of the present study was to disclose the collagen abnormalities in the affected and control fascias and in the patient's fibroblast cultures derived from the skin and fascia. METHODS: The study was performed by histologic, immunohistochemical, and electronmicroscopic techniques. Immunohistochemical studies were done with the use of monoclonal antibodies: anti-collagens I, III, IV, and VI, anti-laminin, anti-fibronectin, anti-desmin, anti-spectrin, anti vimentin, anti-laminin, anti-heparan sulfate, and anti-alpha-actinin. Electronmicroscopic studies were performed on the fascia sections and on cultured fibroblasts. RESULTS: The main abnormality leading to giant collagen fibril formation was presence of myofibroblasts, absence of collagen III, and overproduction of spectrin and collagen type VI, mainly its filamentous form. CONCLUSION: Our findings suggest that the abnormal composition of the fascia could depend on modulation of fibroblasts into myofibroblasts capable of producing spectrin and long-spacing collagen. PMID- 11050584 TI - The burden of psoriasis: a study concerning health-related quality of life among Norwegian adult patients with psoriasis compared with general population norms. AB - BACKGROUND: The reduction of disability caused by psoriasis is an important issue in dermatology. It is thus important to assess the patients' appraisal of their health-related quality of life. OBJECTIVE: The aim of the present study was to assess health-related quality of life among patients with psoriasis and to compare these estimates with population norms. METHODS: The samples comprised 283 patients and 2323 control subjects representative of the general Norwegian population. Health-related quality of life was assessed by means of the SF-36. RESULTS: Both demographic and clinical variables, such as age, gender, educational level, hospital setting, comorbidity, and physical symptoms, affected the different SF-36 scales among the patients. After adjustments had been made for age, gender, and educational level, it was seen that psoriasis patients reported significantly lower scores than the normal controls on all scales. The greatest difference was found on the role emotional scale. The smallest difference was found on the health transition scale. CONCLUSION: These results show that psoriasis patients report poorer health-related quality of life than the general population. Therefore patient care of psoriasis must give attention to the impact of the disease on different life domains. PMID- 11050585 TI - A comparison of hourly block appointments with sequential patient scheduling in a dermatology practice. AB - BACKGROUND: There is significant demand for dermatologic care, and manpower is limited. Increasing patient encounters stress office processes. Analyses of the effects of schedule manipulation in a high-volume dermatology office have not been described. OBJECTIVE: The purpose of this article is to study the effects of block versus sequential scheduling on patient waiting times, length of patient encounters, and physician patient-free time in two busy dermatology clinics. METHODS: A dermatologist attended at two dermatology clinics, one using sequential patient scheduling and the other, block hourly scheduling. Time of patient arrival, scheduled appointment time, waiting time, time of physician entry into the examining room, face-to-face time with the physician, appointment type, number of same-day cancellations and nonattendance by visit type, length of each clinic, time of clinic closure, and patient-free time were recorded for each clinic and patient encounter. RESULTS: There were no significant differences between patient waiting times at the two clinics. Patients were seen by the physician a mean of 2.6 minutes before their scheduled appointment time using block scheduling of appointments and 6 minutes after their scheduled appointment times with sequential scheduling. Similar times were spent by the physician with the patients at both sites. After adjustment for differing nonattendance rates, block scheduling yielded 330 minutes of additional patient-free time during the course of this study when compared with sequential scheduling. With block scheduling, the clinic finished a mean of 35 minutes earlier than clinics using sequential scheduling. CONCLUSION: Within the parameters of this study, block scheduling did not significantly affect patient waiting times. Block scheduling created more patient-free time for the physician and clinical staff than did sequential scheduling. Block scheduling increased the quality of the practice environment from the perspective of the physician and the staff. PMID- 11050586 TI - What is normal black African hair? A light and scanning electron-microscopic study. AB - BACKGROUND: The hair of normal black Africans forms a mat of tightly interwoven hair shafts. The effect of this on the structure of the hair shaft and the response to grooming is unknown. OBJECTIVE: Our purpose was to use light and scanning electron microscopy (SEM) to examine the structure of Negroid-type hairs and effects of combing in black African volunteers. METHODS: Hair samples were collected, by combing, from Africans and compared with those from Caucasian and Asian volunteers. The volunteers had never used chemical treatments. Their hair had not been cut for at least 1 year and grooming had been limited to shampooing, drying, and combing. RESULTS: More than 2000 hairs in 12 African volunteers were examined by light microscopy. The hairs appear as a tight coiled springlike structure. Many shafts exhibited knots (10%-16% vs 0.15%) and appear broken compared with hair shafts from other ethnic groups. SEM of African hairs showed features consistent with repeated breaks of the shaft. Examination of hairs in situ showed interlocking of hair shafts. CONCLUSION: These observations provide an understanding of the physical nature of, and effect of combing on, black African hair. PMID- 11050587 TI - Tazarotene plus UVB phototherapy in the treatment of psoriasis. AB - BACKGROUND: The addition of oral retinoids to phototherapy may accelerate and enhance antipsoriatic efficacy, but can result in systemic adverse events and additional laboratory monitoring costs. OBJECTIVE: Our purpose was to determine whether the topical addition of tazarotene to UVB phototherapy improves efficacy without problems related to photosensitivity. METHODS: Bilateral target plaques were randomized to receive two of the following, one on each plaque once daily for 14 days: tazarotene 0.1% gel, vehicle gel, or no treatment. Thereafter, the same treatments were continued 3 times per week, plus UVB phototherapy 3 times per week, for an additional 67 days. RESULTS: Tazarotene plus UVB phototherapy achieved faster and significantly greater reductions in plaque elevation and scaling throughout treatment and achieved at least 50% improvement from the pretreatment baseline with a significantly lower median cumulative UVB exposure than vehicle gel plus UVB light or UVB phototherapy alone. No case of unusual photosensitivity was noted in the tazarotene plus UVB treatment group. CONCLUSION: The addition of tazarotene to UVB phototherapy improves and accelerates efficacy and maintains acceptable safety and tolerability. PMID- 11050589 TI - Alar batten cartilage grafting in nasal reconstruction: functional and cosmetic results. AB - BACKGROUND: Alar batten cartilage grafts can restore form and function to a compromised ala, prevent stenosis of the nasal valve, and maintain unrestricted air movement. Soft tissue reconstructive options can be combined with alar batten grafts. OBJECTIVE: Our purpose was to analyze functional and cosmetic outcomes in a series of patients undergoing alar batten cartilage grafting. METHODS: We analyzed the functional and cosmetic outcomes of 25 patients in whom reconstruction involved alar batten cartilage grafts. Assessment included defect characteristics, function and cosmesis (rated by physician and patient), and complications. RESULTS: Eighty-three percent of patients had good to excellent functional and cosmetic results by patient and physician assessment. Three patients were rated as having poor cosmetic results by the physician; all 3 patients graded these results as good. One episode of graft failure occurred, and recipient and donor site complications were minor. CONCLUSION: Alar batten cartilage grafts appear to be an excellent option for reconstruction of substantial alar defects. PMID- 11050588 TI - Efficacy of ajoene in the treatment of tinea pedis: a double-blind and comparative study with terbinafine. AB - Ajoene, an organosulfur compound originally isolated from garlic, has been shown to be effective in short-term treatment of tinea pedis. We compare the safety and effectiveness of twice-daily topical application during 1 week of 0.6% and 1% ajoene and 1% terbinafine in the treatment of tinea pedis. Seventy soldiers from the Venezuelan Armed Forces, with clinical and mycologic diagnosis of tinea pedis, were included in this study. However, only 47 were available for final evaluation. The patients were randomly distributed into 3 treatment groups: 0.6% ajoene, 1% ajoene, and 1% terbinafine. Clinical follow-up shows a rapid decline in the signs and symptoms in all groups. Efficacy of the treatments, measured as mycologic cure, 60 days after the end of the therapy was 72% for 0.6% ajoene, 100% for 1% ajoene, and 94% for 1% terbinafine. This represents the first demonstration of the therapeutic application of an inhibitor of phospholipid biosynthesis in human dermatophytosis. PMID- 11050590 TI - Nasal reconstruction utilizing a muscle hinge flap with overlying full-thickness skin graft. AB - BACKGROUND: Deep nasal defects of the dorsum, sidewall, and ala can be challenging to repair. OBJECTIVE: The article describes our experience with a muscle hinge transposition flap with overlying local full-thickness skin grafting for repair of deep nasal defects in a single-stage procedure. METHODS: A muscle hinge transposition flap with overlying local full-thickness skin grafting was used immediately after Mohs micrographic surgery to repair 12 deep nasal defects of the dorsum, sidewall, alar lobule, and supratip. RESULTS: No cases of infection, flap, or graft necrosis occurred in our series. Cosmetic and functional outcomes were judged from good to excellent by patient and surgeon. To enhance the cosmetic outcome, 5 patients underwent spot dermabrasion within 2 months of repair. CONCLUSION: For properly selected small to medium-sized deep nasal defects (1-2 cm) that lack a sufficiently loose adjacent tissue reservoir for a single-stage local flap, a muscle hinge transposition flap with local full thickness skin grafting can provide consistently satisfying aesthetic and functional results. PMID- 11050591 TI - Erythromelalgia: new theories and new therapies. AB - Erythromelalgia is a rare condition that has remained an enigma diagnostically and therapeutically for decades. It has been assumed that erythromelalgia, which is characterized by hot, red, intensely painful feet or hands, may be the opposite of Raynaud's phenomenon. However, new research suggests that these two disorders are more similar than dissimilar. Erythromelalgia usually follows a chronic, sometimes progressive and disabling course. New evidence suggests that this may not be a disease entity at all, but a syndrome of dysfunctional vascular dynamics; recent studies demonstrate that this dysfunction is reversible in some patients. This review article presents the latest theories and successful treatments for erythromelalgia, and data from a survey of members of The Erythromelalgia Association, which was formed to provide information about erythromelalgia to doctors and patients. PMID- 11050592 TI - Psychodermatology. PMID- 11050593 TI - Surgical Pearl: curetting with a punch. PMID- 11050594 TI - Treatment of Birt-Hogg-Dube syndrome with erbium:YAG laser. AB - We describe a female patient with Birt-Hogg-Dube syndrome presenting with multiple, disfiguring papules on the cheeks, nose, and ears. In addition, there was evidence of photoaggravation and inflammatory processes in this case. The patient was treated with an erbium:YAG laser. Although the lesions could be successfully ablated up to the papillary dermis without scar formation, a relapse was observed after 6 months. Thus deeper laser ablations may possibly prevent relapses from dermal residual lesions. PMID- 11050595 TI - Sustained improvement of the quality of life of patients with psoriasis after hospitalization. AB - Psoriasis is a chronic, recurrent, and often disfiguring skin disease that may significantly affect patients' quality of life. Treatment of psoriasis, including hospitalization, has been shown to improve quality of life. A pilot study of 15 consecutive inpatients and 7 consecutive outpatients with psoriasis were asked to complete the Dermatology Life Quality Index (DLQI) before treatment and 3 months later. Hospitalized patients also completed the DLQI 1 week after discharge. Statistical analysis using t tests compared pretreatment and posttreatment DLQI scores as well as improvement of inpatients versus outpatients. Baseline DLQI scores for hospitalized patients were significantly higher (greater impairment of life quality) compared with oupatients' quality of life. After discharge, hospitalized patients' quality of life had significantly improved at 1 week and remained improved at 3 months. PMID- 11050596 TI - Mycosis fungoides d'emblee: CD30-negative cutaneous large T-cell lymphoma. AB - The d'emblee variant of mycosis fungoides initially described patients with a rapid onset of tumors without progression through patch- and plaque-stage disease. We report a case of a patient with the clinical presentation of mycosis fungoides d'emblee and correlate the histologic and immunophenotypic data with those of a more updated classification scheme. PMID- 11050597 TI - Ipsilateral renal dysplasia with hypertensive heart disease in an infant with cutaneous varicella lesions: an unusual presentation of congenital varicella syndrome. AB - A child with congenital varicella syndrome including cutaneous lesions and ipsilateral renal dysplasia with hypertensive heart disease is described. Varicella was contracted during the tenth week of gestation. Typical congenital varicella bullae, high titer of anti-varicella-zoster virus IgM, and a small right kidney were noted after birth. Hypertensive heart disease resulting from renal dysplasia occurred at 1 year of age. The cutaneous lesions and the dysplastic kidney involved the same dermatomes. Nephrectomy proved to be the treatment of choice for hypertension and congestive heart failure. PMID- 11050598 TI - Multiple pseudolymphomas caused by Hirudo medicinalis therapy. AB - Therapy with medicinal leeches (Hirudo medicinalis) is now frequently applied in plastic surgery and in the management of chronic venous insufficiency. We observed a patient in whom firm, brown-red, pea-sized papules developed at each site where leeches had been applied on the lower legs. Histology, immunohistology, and molecular analysis of T-cell receptor and immunoglobulin heavy chain gene rearrangement proved these lesions to be follicular pseudolymphomas. PMID- 11050599 TI - Neutrophilic dermatosis of the dorsal hands: pustular vasculitis revisited. AB - An entity termed "pustular vasculitis of the hands" was recently described. Patients with this condition presented with low-grade fevers and erythematous plaques, pustules, and bullae limited to the dorsal hands and fingers, which were characterized histologically by a dense neutrophilic infiltrate and leukocytoclastic vasculitis. We describe patients with a similar clinical presentation, but who lacked vasculitis on biopsy findings. We describe 3 otherwise asymptomatic patients with hemorrhagic bullae, plaques, and pustules solely on the dorsal hands. Biopsy specimens showed a neutrophilic infiltrate and leukocytoclasis, but no necrotizing vasculitis, and were reminiscent of Sweet's neutrophilic dermatoses. In our patients, corticosteroids or dapsone led to clearing of the lesions, and small maintenance doses of dapsone prevented their recurrence. Our 3 patients had clinical lesions similar to those termed pustular vasculitis of the hands, but which lacked leukocytoclastic vasculitis on biopsy findings. Because of histologic findings and a therapeutic response more characteristic of Sweet's syndrome, we propose the term neutrophilic dermatosis of the dorsal hands. In addition, low-dose dapsone is proposed as a possible first-line therapy in this condition, especially in those with recurrent disease. PMID- 11050600 TI - Losing touch with the healing art: dermatology and the decline of pastoral doctoring. AB - Technological advance in society and medicine has brought tremendous improvements and convenience but also a degree of depersonalization. The personal and pastoral aspects of medical practice, which are probably more important in helping patients toward health than we realize, are becoming increasingly stifled by health care systems which are increasingly "scientific," technological, and "efficient." Clinical practice in dermatology requires pastoral as well as technical skills, art as well as science, and yet the balance of current medical culture increasingly favors and encourages "science" over "art." In dermatology, this bias is evident in a reductionist focus of research, the move towards evidence-based medicine and the emergence of teledermatology. Although all these developments are extremely important and valuable, their effect on the doctor patient relationship needs to be considered carefully. Increasingly rapid scientific advance is paradoxically providing diminishing returns for patients and the healing art is still very much in demand. PMID- 11050601 TI - Time alone, Y2K, and for-profit medicine. PMID- 11050602 TI - Ultraviolet A1 (340-400 nm) phototherapy for cutaneous T-cell lymphoma? PMID- 11050603 TI - Actinic keratosis is squamous cell carcinoma. PMID- 11050604 TI - Terbinafine and potential drug interactions. PMID- 11050606 TI - Helicobacter pylori and rosacea. PMID- 11050607 TI - Milton Orkin (1929-1999). PMID- 11050608 TI - In vivo transient vibration assessment of the normal human thoracolumbar spine. AB - OBJECTIVE: The objective of this study was to quantify the mobility characteristics (dynamic stiffness and mechanical impedance) of the normal human thoracolumbar spine with a transient vibration analysis technique. DESIGN: This study is a prospective clinical investigation to obtain normative biomechanical data from the human male and female spine in vivo. SETTING: Musculoskeletal research laboratory, university setting. SUBJECTS: Twenty asymptomatic subjects (age range, 20-60 years) with no recent history of musculoskeletal complaints. MAIN OUTCOME MEASURES: Mechanical impedance, effective stiffness, and resonant frequency analyses were used to quantify the dynamic stiffness of the thoracolumbar spine in this subject population. Data were obtained from posteroanterior mechanical thrusts delivered with an activator adjusting instrument equipped with a load cell and accelerometer by means of a portable computer. RESULTS: In response to the activator adjusting instrument thrusts, the thoracolumbar spine typically exhibited an impedance minimum at frequencies ranging between 30 and 50 Hz. The maximum posteroanterior impedance and corresponding maximum effective stiffness of the thoracolumbar spine and sacrum was roughly 2 to 8 times greater than the magnitude of the impedance minimum. Statistically significant differences in mobility between male and female subjects were noted, particularly for frequencies corresponding to the maximum mobility (40 Hz) and minimum mobility (10-20 Hz, 70-80 Hz). For most subjects (both male and female), the lumbar region exhibited a higher impedance and stiffness (less mobility) when compared with the thoracic region. CONCLUSIONS: The posteroanterior mechanical behavior of the human thoracolumbar spine was found to be sensitive to mechanical stimulus frequency and showed significant region-specific and gender differences. In the frequency range of 30 to 50 Hz, the lumbar spine of this subject population is the least stiff and therefore has the greatest mobility. From a biomechanical point-of-view, the results of this study indicate that dynamic spinal manipulative therapy procedures will produce more spinal motion for a given force, particularly when the posteroanterior manipulative thrust is delivered in frequency ranges at or near the resonant frequency. In this regard, spinal manipulative therapy procedures designed to target the resonant frequency of the spine require less force application. Both magnitude and frequency content of manual and mechanical thrusting manipulations may be critical elements for therapeutic outcome. PMID- 11050609 TI - Highlighting of intervertebral movements and variations of intradiskal pressure during lumbar spine manipulation: a feasibility study. AB - OBJECTIVES: To demonstrate relative movement of the vertebrae and variations in intradiskal pressure during 2 different lumbar spinal manipulations, in flexion or extension, in 2 unembalmed cadavers. DESIGN: A pressure sensor was inserted into the L3-4 disk in cadaver 1 and into the L1-2 to L4-5 disks in cadaver 2. Two adjacent vertebrae (L3 and L4 in cadaver 1, and L4 and L5 in cadaver 2) were each equipped with 2 monoaxial accelerometers to record acceleration in the caudocranial axis and a biaxial accelerometer to record acceleration in the "horizontal" anatomic plane. SETTING: Laboratory study. RESULTS: During the thrust, relative intervertebral movements were demonstrated; movements differed with the type of manipulation (in flexion or extension). Intradiskal pressure initially increased, then decreased. CONCLUSIONS: Lumbar spinal manipulations have a biomechanical effect on the intervertebral disks, producing a brief but marked change in intradiskal pressure. This effect, which differs slightly with the different types of manipulation studied, is the consequence of movements of the adjacent vertebrae. PMID- 11050610 TI - Nonoperative treatments for sciatica: a pilot study for a randomized clinical trial. AB - OBJECTIVES: To assess the feasibility of patient recruitment, the ability of patients and clinicians to comply with study protocols, and the use of data collection instruments to collect cost-effectiveness data, and to obtain variability estimates for sample-size calculations for a full-scale trial. STUDY DESIGN: Prospective, observer-blinded, pilot randomized clinical trial. SETTING: Primary contact chiropractic and medical clinics. PATIENTS: Ages 20 to 65 years, with low back-related radiating leg pain (sciatica). OUTCOME MEASURES: Self report questionnaires were administered at baseline and 3 and 12 weeks after randomization. The measures included leg and back pain severity, frequency and bothersomeness of symptoms, leg/back disability, medication use, global improvement, satisfaction, and health care utilization. INTERVENTIONS: Medical care, chiropractic care, and epidural steroid injections. RESULTS: A total of 706 persons were screened by phone to determine initial eligibility. Of these, over 90% of those persons contacted did not meet the entrance criteria. The most common reason for disqualification was that the duration of the complaint was longer than 3 months. Twenty patients were randomized into the study. All 3 groups showed substantial improvements in the main patient-rated outcomes at the end of the 12-week intervention phase. For leg pain, back pain, frequency and bothersomeness of leg symptoms, and Roland-Morris disability score, the percent improvement varied from 50% to 84%, and the corresponding effect sizes ranged from 0.8 to 2.2. Bothersomeness of leg symptoms was the most responsive outcome associated with the largest magnitude of effect size. All within-group changes from baseline were statistically significant (P <.01). No between-group comparisons were planned or performed because of the insufficient sample size and high risk of committing type I and type II errors. CONCLUSIONS: Pilot studies such as these are important for the determination of the feasibility of conducting costly, larger scale trials. Recruitment for a full-scale study of sciatica of 2 to 12 weeks duration is not feasible, given the methods used in this pilot study. Our results do indicate, however, that there are substantial numbers of patients with sciatica more chronic in nature who would be interested in participating in a similar study. In addition, collaboration with a local health maintenance organization would likely facilitate clinician referrals and optimize the recruitment process. Patient and provider compliance was high in the pilot study, which indicates that most study protocols are feasible. Additionally, we were able to collect complete outcomes data, including those regarding health care use. A suggested modification by investigators and outside consultants has resulted in the replacement of the medication group with a minimal intervention control group (ie, self-care advice). As a result, a second pilot study of patients with sciatica of more than 4 weeks duration has been planned before a full-scale trial is attempted. PMID- 11050611 TI - Effects of spinal manipulative therapy on autonomic activity and the cardiovascular system: a case study using the electrocardiogram and arterial tonometry. AB - OBJECTIVE: To determine if there is alteration in the autonomic nervous and cardiovascular systems after chiropractic manipulative therapy (CMT). A novel approach was used to quantitatively probe for changes in the activity of the autonomic nervous system, in blood pressure, and in pressure pulse transmission time. This approach uses the electrocardiogram and arterial tonometry equipment. DESIGN: This case study involves 1 subject treated over a 6-week period (2 visits/week). Respiration, electrocardiogram, and left and right radial artery blood pressures were measured during the baseline (2 visits) and treatment (10 visits) phases. Measurements were obtained before (n = 3) and after (n = 3) a break period (baseline) or before and after CMT. High-velocity, low-amplitude CMT that produced joint cavitation was used. SETTING: The study was performed at the Parker College Research Institute in a temperature-controlled laboratory. MAIN OUTCOME MEASURES: Fourier analysis was performed on the electrocardiogram determined rest-redistribution intervals. The low frequency power between 0.04 to 0.15 Hz and the high frequency power between 0.15 to 0.40 Hz represent the activity of the sympathetic and parasympathetic nervous systems, respectively. The main outcome measure was the sympathovagal index, which is determined from the ratio of low frequency to high frequency. The arterial pressure and the time for pressure pulses to travel from the heart to the radial artery recording sites (pressure pulse transmission time) were studied. Differences (average of 3 measurements after treatment minus measurements before treatment) for each variable were calculated. RESULTS: After the 1st CMT treatment, the difference between treatment and baseline decreased for both the low frequency/high frequency (-2.804 +/- 1.273) and low frequency power (-0.135 +/- 0.056). These findings indicated that the parasympathetic nervous system predominated the sympathetic nervous system. After the 3rd, 4th, 6th, and 9th treatment, the difference between treatment and baseline increased for low frequency/high frequency (0.908 +/- 0.338, 2.313 +/- 0.300, 2.776 +/- 1.102, and 0. 988 +/- 0.269, respectively) and indicated that the sympathetic nervous system predominated the parasympathetic nervous system. In addition, the difference between treatment and baseline for the pressure pulse transmission time decreased bilaterally after the 4th treatment (left, -13.52 +/- 3.70 ms; right, -9.75 +/- 3.76 ms) and 6th treatment (left, -9.53 +/- 3.60 ms; right, -9.24 +/- 3.50 ms), which indicated that arterial compliance had decreased. Furthermore, after the 6th treatment, the difference between treatment and baseline for the rest redistribution interval time decreased (-0.084 +/- 0.014 s). The difference between treatment and baseline for the systolic, diastolic, and mean arterial pressure for the above-mentioned treatments was not considered significant. CONCLUSION: This case study is the first to attempt to use electrocardiogram and arterial tonometry data to study the effects of CMT on the autonomic nervous and cardiovascular systems over an extended period of time. These devices allowed a more in-depth study of the cardiovascular and autonomic changes associated with CMT. Although changes in the autonomic nervous and cardiovascular systems can be detected, further development of a reliable and reproducible experimental protocol is required before validating the effects of CMT on these systems. PMID- 11050612 TI - The role of spinal tissues in resisting posteroanterior forces applied to the lumbar spine. AB - OBJECTIVE: To determine the effects of the dissection of spinal tissues on the mechanical behavior of motion segments under the application of posteroanterior forces. DESIGN: A cadaveric motion segment study. SETTING: A tissue mechanics research laboratory. PROCEDURE: Anterior shear and extension moment were applied to 10 motion segments to simulate the clinical situation when posteroanterior forces were applied to the spine. The movements of the specimens in the sagittal plane were studied by a camera. Spinal tissues were dissected sequentially, and the mechanical testing was repeated after the dissection of each tissue. RESULTS: The most significant movements produced were extension and superior translation of the anteroinferior corner of the superior vertebral body. Translational movements in the other directions were small. The dissection of the posterior ligaments and zygapophyseal joints did not lead to significant changes in the movements. CONCLUSIONS: Injuries of the posterior ligaments are unlikely to alter the mechanical response of the spine to posteroanterior forces. However, these posterior tissues are pain sensitive and may be subjected to large strains and elicit symptoms. PMID- 11050613 TI - Chiropractic patients in the Netherlands: a descriptive study. AB - BACKGROUND: Despite the worldwide popularity of chiropractic, there is still relatively little known about the patients who visit chiropractors in the Netherlands and other European countries. OBJECTIVE: To describe in-depth the patient population of new patients to chiropractors in the Netherlands. DESIGN: Study population consisted of 10 consecutive new patients per participating chiropractor. A retrospective-type questionnaire was used. SETTING: Private practice. OUTCOME MEASURES: Mode of referral, area, and nature of the complaints; related to the chief complaint: previous treatments, examinations, type of referral, days lost at work, level of pain, and treatment expectations. RESULTS: Of the 130 chiropractors registered with the Netherlands Chiropractors'Association, 94 chiropractors(78%) participated. Eight hundred thirty-three patients (89%) returned questionnaires. By far, the greatest reason that patients visit chiropractors in the Netherlands is for neuromusculoskeletal (NMS) complaints. At the time of examination, 86% of the patients had spinal related complaints, of which 12% involved multiple areas of the spine. Non-NMS complaints are minimal (<2%). Seventy-seven percent of patients with NMS complaints have chronic complaints (>12 weeks). Three-quarters of these patients have undergone previous conservative therapy for their complaint, which includes physical and manual therapy, postural correction, and exercise therapy. Despite the chronic nature of their complaints, patients have high expectations that their treatment will be effective. CONCLUSIONS: Most patients who see chiropractors in the Netherlands have chronic NMS-related complaints. Chiropractors are not a part of the normal referral system in this country, with the result that the patients have rather long histories, including previous evaluations by medical specialists and other previous forms of (conservative) care. PMID- 11050614 TI - The role of the gamma-motor system in increasing muscle tone and muscle pain syndromes: a review of the Johansson/Sojka hypothesis. AB - OBJECTIVES: To review literature that pertained to the Johansson/Sojka hypothesis that positive feedback loops in the gamma-motor system are responsible for chronic muscle pain and increases in muscle tone. DATA SOURCES: Articles were selected from MEDLINE searches and from manual library searches. RESULTS: Normal, static, and ischemic muscle contractions and/or chemical mediators of inflammation excite intramuscular groups III and IV chemonociceptors. In groups III and IV, afferent firing stimulates gamma-motorneurons, which causes the firing of Ia and II muscle spindle afferents and increased extrafusal resistance to stretch (muscle tone). Some criticism of the involvement of the gamma-motor system in muscle tone was found to be dated or based on data from noncomparable research. Most of these studies (pro and con) were performed on prepared test animals, and the results may or may not translate to human subjects. CONCLUSIONS: There exists a sizable body of research that establishes a link between the activation of intramuscular chemonociceptors, increased gamma-motor activity, and increased Ia and II spindle output, as proposed by the hypothesis of Johansson and Sojka. However, because of the lack of sufficient data on human subjects, their hypothesis should not be considered proved. Further research into the effects of metabolites of muscle contraction and their effects on muscle tone is recommended. Research into subluxation/joint dysfunction in light of the Johansson/Sojka hypothesis is recommended. PMID- 11050615 TI - Upper cervical chiropractic management of a patient with Parkinson's disease: a case report. AB - OBJECTIVE: To discuss the use of upper cervical chiropractic management in managing a single patient with Parkinson's disease and to describe the clinical picture of the disease. CLINICAL FEATURES: A 60-year-old man was diagnosed with Parkinson's disease at age 53 after a twitch developed in his left fifth finger. He later developed rigidity in his left leg, body tremor, slurring of speech, and memory loss among other findings. INTERVENTION AND OUTCOME: This subject was managed with upper cervical chiropractic care for 9 months. Analysis of precision upper cervical radiographs determined upper cervical mis-alignment. Neurophysiology was monitored with paraspinal digital infrared imaging. This patient was placed on a specially designed knee-chest table for adjustment, which was delivered by hand to the first cervical vertebrae, according to radiographic findings. Evaluation of Parkinson's symptoms occurred by doctor's observation, the patient's subjective description of symptoms, and use of the Unified Parkinson's Disease Rating Scale. Reevaluations demonstrated a marked improvement in both subjective and objective findings. CONCLUSION: Upper cervical chiropractic care aided by cervical radiographs and thermal imaging had a successful outcome for a patient with Parkinson's disease. Further investigation into upper cervical injury as a contributing factor to Parkinson's disease should be considered. PMID- 11050616 TI - 9th international workshop on fragile X syndrome and X-linked mental retardation. PMID- 11050617 TI - Syndromic XLMR genes (MRXS): update 2000. PMID- 11050618 TI - MRX review. PMID- 11050619 TI - Rho proteins and the cellular mechanisms of mental retardation. AB - The biological basis of mental retardation is poorly understood. Mental retardation is associated with an immature morphology of synaptic spines, structures involved in neurotransmission and memory processes, suggesting that mental retardation is due to a deficiency in neuronal network formation. Recently, several genes involved in X-linked mental retardation (MRX) have been cloned. Investigation of the roles of these genes in neuronal development and function should lead to a better understanding of the cellular mechanisms underlying mental retardation. A significant number of MRX genes is directly involved in signal transduction through Rho proteins. These Rho proteins act as molecular switches which integrate extracellular and intracellular signals to regulate rearrangement of the actin cytoskeleton. Since the actin cytoskeleton mediates neuronal motility and morphogenesis, one can envision how mutations in proteins involved in Rho-dependent signaling result in mental retardation by altering neuronal network formation. PMID- 11050620 TI - Mechanisms, models, and mental retardation. PMID- 11050621 TI - Mapping to distal Xq28 of nonspecific X-linked mental retardation MRX72: linkage analysis and clinical findings in a three-generation Sardinian family. AB - Families with mentally retarded males found to be negative for FRAXA and FRAXE mutations are useful in understanding the genetic basis of X-linked mental retardation. According to the most recent data (updated to 1999), 69 MRX loci have been mapped and 6 genes cloned. Here we report on a linkage study performed on 20 subjects from a 4-generation Sardinian family segregating a non-specific X linked recessive mental retardation (XLMR)(MRX72) associated with global delay of all psychomotor development. Five of 8 affected males have been tested for mental age, verbal and performance skills and behavioral anomalies; mental impairment ranged from mild to severe. Only minor anomalies were present in the affected subjects. Two-point linkage analysis based on 28 informative microsatellites spanning the whole X chromosome demonstrated linkage between the disorder and markers DXS1073 and F8c in Xq28 (maximum Lod score of 2. 71 at straight theta = 0.00). Multipoint linkage analysis confirmed the linkage with a Z(max) of 3.0 at straight theta = 0.00 at DXS1073 and F8c. Recombination in an affected male at DXS1073 and F8c allowed us to delimit centromerically and telomerically the region containing the putative candidate gene. The region, where MRX72 maps, overlaps that of another MRX families previously mapped to Xq28, two of which harbored mutations in GDI. Involvement of this gene was excluded in our family, suggesting another MRX might reside in Xq28. PMID- 11050622 TI - Holmes-Gang syndrome is allelic with XLMR-hypotonic face syndrome. PMID- 11050623 TI - Exclusion of nine candidate genes for their involvement in X-linked FG syndrome (FGS1) in three families. PMID- 11050624 TI - Molecular study of the PAK3 and GDI1 genes in nonsyndromic X-linked mental retardation spanish patients. PMID- 11050625 TI - Distal 5q trisomy resulting from an X;5 translocation detected by chromosome painting. AB - We describe the case of a 13-year-old girl with an apparently de novo unbalanced translocation resulting in the presence of additional chromosomal material on the short arm of one X chromosome, which was detected by conventional G-banding studies. Fluorescence in situ hybridization (FISH) using the Chromoprobe Multiprobe-M protocol confirmed that the additional chromosomal material originated from chromosome 5. The karyotype of this patient is now established to be 46,X,der(X) t(X;5)(p22.3;q33), with a deletion of Xp22.3-pter and partial trisomy of 5q33-qter. The distal 5q trisomy genotype has been associated with clinical signs that include growth and mental retardation, eczema, craniofacial anomalies, and malformations of heart, lungs, abdomen, limbs, and genitalia. Our patient also has short stature, a prominent nasal bridge, a flat philtrum, a thin upper lip, dental caries, and limb and cardiac malformations, but she appears to be mildly affected compared with previously reported cases. This is the first case of distal 5q trisomy arising from a translocation with the X chromosome. Replication studies on this patient show that the derivative t(X;5) chromosome is late replicating in almost all cells examined, which indicates that this chromosome is preferentially inactivated. However, the translocated segment of chromosome 5 appears to be early replicating, which implies that the trisomic 5q segment is transcriptionally active. We cannot determine from these studies whether all or only some genes in this segment are expressed, but this patient's relatively mild clinical signs suggest that the critical region(s) that contribute to the distal 5q trisomy phenotype are at least partly suppressed. A review of other patients with X-chromosome translocations indicates that many but not all of them also have attenuated phenotypes. The mechanism of inactivation of autosomal material attached to the X chromosome is complex, with varying effects on the phenotype of the patients that depend on the nature of the autosomal chromatin. Replication studies are of limited utility in predicting expression of autosomal genes involved in X-chromosome translocations. PMID- 11050626 TI - Transverse limb defects associated with aorto-pulmonary vascular abnormalities: vascular disruption sequence or atypical presentation of Adams-Oliver syndrome? AB - We report a patient with terminal transverse limb defects associated with persistent primitive aorto-pulmonary vascular connections leading to supra systemic pulmonary artery pressure. It is likely that this patient represents a vascular disruption sequence or as an alternative a form of Adams-Oliver syndrome. These assumptions are based only on the association of vascular abnormalities as an emerging and apparently important association with transverse limb defects despite the absence of aplasia cutis congenita commonly associated with Adams-Oliver syndrome. PMID- 11050627 TI - Gomez-Lopez-Hernandez syndrome: expansion of the phenotype. AB - Gomez-Lopez-Hernandez syndrome (cerebello-trigeminal-dermal dysplasia) is a condition that includes abnormalities of the cerebellum (rhombencephalosynapsis), cranial nerves (trigeminal anesthesia), and scalp (alopecia). Seven patients with this condition have been documented since 1979. We now report a male with Gomez Lopez-Hernandez syndrome who, at the age of 19 years, is the oldest patient identified to date. He has been followed since birth, allowing us to report on the progression of his physical findings and psychiatric problems including hyperactivity, depression, self-injurious behavior and bipolar disorder. In addition, he has short stature and growth hormone deficiency. PMID- 11050628 TI - Primary care physicians' perceptions of barriers to genetic testing and their willingness to participate in research. AB - Our objective was to explore the barriers and motivations to: 1) appropriate diffusion of genetic services into primary care practice; and 2) primary care physicians' (PCPs) willingness to participate in clinical studies to assess the safety and effectiveness of emerging genetic technologies. A random sample (n = 994) of PCPs was invited to be interviewed. Of the 80 who agreed, 60 were interviewed, 52 by telephone. A semi-structured guide was used. A questionnaire mailed to 752 of the PCPs was used to elicit information from physicians who did not want to be interviewed. Among interviewees, uncertainty as to the clinical utility and clinical validity of predictive genetic testing were the leading barriers to incorporation of this technology into practice, being mentioned by 60 and 43% of subjects, respectively. Of the 100 (13. 3%) physicians returning the questionnaire who declined to be interviewed, 30% said they would be willing to participate in research on the safety and effectiveness of predictive genetic tests. Of those who were interviewed, 92% were willing to participate in such research. Most physicians do not see genetics as important in their practice today; many anticipate greater importance in the future. The proportion of physicians interested in participating in research to assess the safety and effectiveness of genetic tests is sufficient to make large scale, collaborative, practice-based evaluation feasible. Additionally, participation in research may serve as an effective medium for physician education in genetics. PMID- 11050629 TI - Mutation analysis of Gaucher disease using dot-blood samples on FTA filter paper. AB - FTA((R)) filter papers were used as an effective means of blood cell collection, genomic DNA processing, and delivery. Minute blood samples (<1 microL) were collected onto the filters via a simple lateral prick to the patient's finger, circumventing the need for intravenous blood puncture. Collected samples, which are stable at room temperature for several years, were subsequently sent through the postal system to the diagnostic laboratory, bypassing the stringent requirements of courier delivery. Using this method, we performed restriction fragment length polymorphism (RFLP) and nucleotide sequence analysis on prevalent mutations among Canadian and Chinese Gaucher disease patients. Of the 12 alleles (six patients) analyzed, 42% (5/12) have the N370S mutation and 58% (7/12) the L444P mutation, the two most common alleles found among Jewish and non-Jewish Gaucher disease patients. Uniquely, a Chinese Gaucher disease patient was found to have an N370S mutation. Although the presence of the N370S mutation is regarded as common in other ethnic groups, previous to this report it had not been noted in an individual of Asian descent. PvuII polymorphism analysis showed that the N370S mutation found in the Chinese patient was linked to the Pv1.1(-) polymorphism, as has been previously seen in the Jewish population. The use of FTA((R)) filter paper facilitates access of samples to diagnostic centers, and therefore provides an effective means of performing population-based mutational analysis of Gaucher disease internationally. PMID- 11050630 TI - Autosomal dominant inheritance in Cantu syndrome (congenital hypertrichosis, osteochondrodysplasia, and cardiomegaly). AB - Cantu syndrome (CS) is characterized by congenital hypertrichosis, osteochondrodysplasia, cardiomegaly, and coarse facial appearance; autosomal recessive inheritance has been postulated. We report on a Mexican family with CS; the affected members are the 44-year-old father and his two children (a male and female), aged 14 and 4 years, respectively; each shows the classic characteristics, but the father and the brother also have a previously unreported feature, namely, a thick calvarium. This is the first reported instance of male to-male transmission of CS. With the paternal age effect found in the reported sporadic cases and the segregation analysis [Robertson et al., 1999], autosomal dominant inheritance is more likely than autosomal recessive inheritance. The cases of affected sibs reported by Cantu et al. [1982] could be explained by parental gonadal mosaicism. PMID- 11050631 TI - Phenotype of a patient with pure partial trisomy 2p(p23-->pter). AB - We present the case of a 7-month-old girl with the karyotype 46,XX, der(13) t(2;13)(p23;p11.2).ish der(13)(wcp2+) de novo. Painting confirmed that the additional segment on 13p was of chromosome 2 origin, resulting in trisomy 2p23 - >2pter. The child had a prominent forehead with a flat hemangioma, depressed nasal bridge, protruding tongue, posteriorly angulated ears, esotropia with poor abduction of the right eye, bilateral severe myopia (-5.5 D), retinal hypopigmentation, foveal hypoplasia, and striking left optic nerve hypoplasia. She also had pectus excavatum, a protruding abdomen with diastasis recti, generalized hypotonia, delayed fine and gross motor development, grade II reflux on the left side, and grade III-IV reflux on the right side. An EEG showed epileptiform discharges. Computed tomographic scan of the brain showed decreased white matter, but magnetic resonance imaging showed normal results. PMID- 11050632 TI - Bardet-Biedl syndrome type 3 in an Iranian family: clinical study and confirmation of disease localization. AB - Bardet-Biedl syndrome (BBS) is a group of autosomal recessive MCA/MR syndromes characterized by pigmentary retinopathy, postaxial polydactyly, hypogenitalism, obesity, and mental retardation. Five BBS loci have been identified; among them, BBS type 1 (BBS1) and type 3 (BBS3) are most common and most rare, respectively. We encountered an Iranian family that had seven affected members. All patients had a history of mild to severe obesity, but it was reversible in some patients by caloric restriction and exercise. All patients had pigmentary retinopathy, beginning as night blindness in early childhood and progressing toward severe impairment of vision by the end of the second decade. Polydactyly varied in limb distribution, ranging from four-limb involvement to random involvement or even to nonaffectedness. Six of the seven patients were not mentally retarded. Although kidney anomaly or an adrenal mass was pres- ent in two patients, the fact that one patient had seven children rules out reproductive dysfunction. Linkage analysis with microsatellite markers showed that the disease in the family was assigned to a region around marker loci at 3p13-p12 (maximum LOD score = 4.15 and recombination fraction straight theta = 0, at D3S1603 microsatellite marker), to which the BBS3 locus has been mapped. Haplotype analysis did not reduce the extent of the previously reported critical region of BBS3. A comparison of clinical manifestations of our patients with those of previously reported BBS3 patients did not support any type-specific phenotypes, though manifestations in our patients are similar to those in BBS3 patients of a family in Newfoundland. PMID- 11050633 TI - Erratum: finckh U, Schroder J, ressler B, veske A, gal A. Spectrum and detection rate of L1CAM mutations in isolated and familial cases with clinically suspected L1-disease. Am J med genet 92:40-46 PMID- 11050634 TI - Staging of rectal cancer by double-contrast MR imaging using the rectally administered superparamagnetic iron oxide contrast agent ferristene and IV gadodiamide injection: results of a multicenter phase II trial. AB - The aim of this study was to assess the accuracy of double-contrast magnetic resonance imaging (MRI) with rectal application of the superparamagnetic iron oxide contrast agent (SPIO) ferristene and IV gadodiamide for preoperative staging of rectal cancer. In a randomized phase II dose-ranging trial, 113 patients were studied preoperatively with one of four different formulations of ferristene (Abdoscan) as an enema before MRI. T1-weighted spin-echo (T1w SE) and T2w turbo spin-echo (TSE) single-contrast images were obtained as well as T1w SE and gradient-echo (GRE) double-contrast images after IV gadodiamide injection (Omniscan). Images were assessed qualitatively, and TNM tumor stage was compared with histopathology. High-viscosity ferristene formulations were superior to low viscosity formulations in tumor staging (accuracy 90% vs 74%, P < 0.01). There was no significant difference between high and low iron content ferristene. MRI had a sensitivity of 97%, specificity of 50%, and accuracy of 82% for staging of rectal carcinoma higher than T2 stage. At receiver operator characteristic (ROC) analysis, MR differentiation between T1/T2 and T3/T4 tumor stages yielded a ROC index of 0.848. Double-contrast MRI is an accurate method for preoperative staging of rectal cancer. PMID- 11050635 TI - Artery and vein separation using susceptibility-dependent phase in contrast enhanced MRA. AB - In magnetic resonance angiography, contrast agents are frequently used to help highlight arteries over background tissue. Unfortunately, enhancing veins hamper the visualization of arteries when data are collected over a long period of time after the arterial phase of the contrast agent. To overcome this problem, we have developed a novel imaging and postprocessing method that is capable of eliminating veins by utilizing the susceptibility difference between veins and surrounding tissue. This method was applied in the peripheral vasculature where the vessels are predominantly parallel to the main field and where the blood oxygen level-dependent effect is most pronounced. Results are presented for both long (15.8 msec) and short echo times (7.8 msec) and for sequential and centrally reordered acquisition schemes. The short echo scan approach appears to be the most promising, making it possible to obtain good suppression of the venous signal even when the timing is not perfect or when repeat scans are necessary. PMID- 11050636 TI - Contrast-enhanced 3D MRA using SENSE. AB - Sensitivity encoding (SENSE) was used to improve the performance of three dimensional contrast-enhanced magnetic resonance angiography (3D CE-MRA). Utilizing an array of receiver coils for sensitivity encoding, the encoding efficiency of gradient-echo imaging was increased by factors of up to three. The feasibility of the approach was demonstrated for imaging of the abdominal vasculature. On the one hand, using a SENSE reduction factor of two, the spatial resolution of a breath-hold scan of 17 seconds was improved to 1.0 x 2.0 x 2.0 mm(3). On the other hand, using threefold reduction, time-resolved 3D CE-MRA was performed with a true temporal resolution of 4 seconds, at a spatial resolution of 1.6 x 2.1 x 4.0 mm(3). CE-MRA with SENSE was performed in healthy volunteers and patients and compared with a standard protocol. Throughout, diagnostic quality images were obtained, showing the ability of sensitivity encoding to enhance spatial and/or temporal resolution considerably in clinical angiographic examinations. PMID- 11050637 TI - Performance of QRS detection for cardiac magnetic resonance imaging with a novel vectorcardiographic triggering method. AB - In many cardiac patients, image quality and/or scan efficiency is reduced due to imprecise R-wave ability to trigger the scan due to noise on the electrocardiogram (ECG) caused by the magnetic resonance (MR) environment. We developed a triggering system that uses the spatial information of the vectorcardiogram (VCG) to minimize the effects of MR-related noise on triggering. Fifteen volunteers underwent standard cardiovascular MR exams, and a total of 52,474 R-waves were evaluated with the algorithm, giving a performance index of 99.91%. The mean propagation delay of the system was -10.64 +/- 3.19 msec, which falls within the real-time definition for cardiac MRI triggering. Five patients had arrhythmias consisting of premature ventricular depolarizations (PVDs) and supraventricular extra systoles. For those patients with PVDs, all arrhythmic beats were rejected unless they passed through the algorithm's reference point. The performance index for the arrhythmic patients approached 100%. VCG-based triggering has been demonstrated to provide near 100% triggering performance during cardiac MR examinations. PMID- 11050638 TI - Efficacy and safety of mangafodipir trisodium (MnDPDP) injection for hepatic MRI in adults: results of the U.S. Multicenter phase III clinical trials. Efficacy of early imaging. AB - The efficacy of contrast-enhanced magnetic resonance imaging (MRI) for detecting and characterizing, or excluding, hepatic masses was assessed in 404 patients, following the intravenous administration of mangafodipir trisodium (MnDPDP) injection, a hepatic MRI contrast agent. An initial contrast-enhanced computed tomography (CT) examination was followed by unenhanced MRI, injection of MnDPDP (5 micromol/kg IV), and enhanced MRI at 15 minutes post injection. Agreement of the radiologic diagnoses with the patients' final diagnoses was higher for enhanced MRI and for the combined unenhanced and enhanced MRI evaluations than for unenhanced MRI alone or enhanced CT using the clinical diagnosis as the gold standard. Mangafodipir-enhanced MRI uniquely provided additional diagnostic information in 48% of the patients, and patient management was consequently altered in 6% of the patients. MnDPDP-enhanced MRI was comparable or superior to unenhanced MRI and enhanced CT for the detection, classification, and diagnosis of focal liver lesions in patients with known or suspected focal liver disease. PMID- 11050639 TI - The clinical value of ferric ammonium citrate: a positive oral contrast agent for T1-weighted MR imaging of the upper abdomen. AB - This study was undertaken to determine whether ferric ammonium citrate (FAC), a positive magnetic resonance (MR) contrast agent, is of clinical value in demonstrating or excluding pathology of the upper gastrointestinal tract. A retrospective review was performed of pre- and post-FAC studies of MR examinations in 203 patients from phase II and III clinical trials in whom final diagnoses had been established based on the results of biopsy, surgery, or independent imaging procedures. Two independent reviewers made randomized and blinded assessments of the stomach, duodenum, and pancreas. FAC significantly increased the certainty of diagnosis for normal studies of the stomach and duodenum for both readers (P < 0.001) and for abnormal studies of the stomach for one reader (P = 0.004). FAC also significantly increased the certainty of diagnosis for normal pancreas for one reader (P < 0.001). FAC significantly (P < 0.001) increased accuracy and specificity for diagnoses involving the stomach and duodenum for both readers and for one reader for the pancreas. There was significant improvement in sensitivity for gastric diagnoses (P = 0.013) for one reader but not for the duodenum or pancreas. We conclude that FAC is helpful in demonstrating and excluding upper gastrointestinal pathology on MR. PMID- 11050640 TI - Comparison of the hemodynamic response to different visual stimuli in single event and block stimulation fMRI experiments. AB - Experiments with three different types of basic visual stimulation were performed to compare cortical activation in single-event and block trials. Independent of the stimulation paradigm, the single-event presentation leads to highly consistent signal responses regarding both the activated cortical areas and the dynamics of the signal time course. In contrast, signal time courses during block paradigms depend on the stimulus applied and are a complex and nonlinear function of the single-event responses. Additionally, the initial dip during the first 2 seconds after stimulus onset is consistently observed. However, the small amplitude change (-0.1% to -0.3%) requires signal averaging to establish statistical significance of the effect. Furthermore, different patterns of activation were observed within the primary visual cortex. In an anterior part of the primary visual cortex, activation was only observed at the onset and at the cessation of stimulation involving luminance changes. PMID- 11050641 TI - Numerical tissue characterization in MS via standardization of the MR image intensity scale. AB - Image intensity standardization is a recently developed postprocessing method that is capable of correcting the signal intensity variations in MR images. We evaluated signal intensity of healthy and diseased tissues in 10 multiple sclerosis (MS) patients based on standardized dual fast spin-echo MR images using a numerical postprocessing technique. The main idea of this technique is to deform the volume image histogram of each study to match a standard histogram and to utilize the resulting transformation to map the image intensities into standard scale. Upon standardization, the coefficients of variation of signal intensities for each segmented tissue (gray matter, white matter, lesion plaques, and diffuse abnormal white matter) in all patients were significantly smaller (2.3-9.2 times) than in the original images, and the same tissues from different patients looked alike, with similar intensity characteristics. Numerical tissue characterizability of different tissues in MS achieved by standardization offers a fixed tissue-specific meaning for the numerical values and can significantly facilitate image segmentation and analysis. PMID- 11050642 TI - A quantitative analysis of cortical spreading depression events in the feline brain characterized with diffusion-weighted MRI. AB - Cortical spreading depression (CSD) in the gyrencephalic cat brain was detected with diffusion-weighted echoplanar (DWEP) magnetic resonance imaging (4-8/min for 1-2 hours) using a horizontal imaging plane through the suprasylvian (SG) and marginal gyri. A t-statistic mapping technique allowed a quantitative characterization of the passage of events through single-image pixels (0.15 mm(2)), thus providing a resolution unavailable to previous studies in which time dependent changes instead were derived from averaging data over relatively large ROIs. Using the enhanced analysis, CSD events initiated by KCl could be quantified for the first time as primary or secondary according to their spatial and temporal features. Primary events covered 26.2 +/- 9.9 mm(2)of cortical surface (mean +/- SD, n = 7 experiments) and propagated rapidly (3.5 +/- 0.65 mm * min(-1)) with a hemispherical geometry. In contrast, the subsequent secondary events were multiple, spatially restricted (covering 7.6 +/- 4.6 mm(2), P < 0.005), slower in propagation (2.6 +/- 0.41 mm * min(-1), P < 0.012), and often confined to the originating gyrus (26 out of 59 events). However, both event types were associated with significantly reduced apparent diffusion coefficients (ADCs; from 800 to approximately 660 x 10(-6) mm(2)* s(-1), P < 0.05) that were similar for both primary (21 +/- 5.1%) and secondary waves (18 +/- 7. 7%) and that had similar durations (full width at half-maximal height: 86 +/- 17 vs. 79 +/- 20 seconds, respectively). These findings associate CSD for the first time with two categories of ADC disturbance that are similar in amplitude and duration but that differ in spatial extent, velocity, and extensiveness of spread. PMID- 11050643 TI - MR lymphography using iron oxide nanoparticles in rats: pharmacokinetics in the lymphatic system after intravenous injection. AB - The objective of the study was to quantify the kinetics of the superparamagnetic nanoparticle ferumoxtran (AMI 227, Sinerem(R), Combidex(R)) in the efferent lymph of the subdiaphragmatic lymph nodes and in various node groups of the rat to elucidate the uptake mechanism. The thoracic lymph duct was catheterized in 24 rats after an IV injection of 40 micromol Fe/kg ferumoxtran. Three rats were studied at several time points between 1.5 and 24 hours. At each time point, 0.3 ml of lymph were collected over 45 minutes. Lymph nodes were differentiated into five groups. The iron concentration in the samples and in plasma was measured by relaxometry at 0.47 T and atomic absorption spectrometry. Cytology was performed on the lymph. High concentrations of nanoparticles were found in the thoracic lymph soon after injection (90 minutes). No particle was found in the lymph cells, indicating that ferumoxtran was extracellular in the lymph fluid. The maximum concentration was reached later in all node groups, at 12 hours, and then plateaued. The transcapillary pathway and subsequent lymph drainage of the particles seem to play a major role in the delivery to the lymph nodes. PMID- 11050645 TI - Imaging of urea using chemical exchange-dependent saturation transfer at 1.5T. AB - The purpose of this study was to screen for slow proton chemical exchange between water and kidney metabolites using a standard clinical 1.5-T scanner. Imaging was performed using a fast spin-echo sequence with a magnetization transfer (MT) preparation pulse train. Off-resonance saturation ranging from +/-50 to +/-1000 Hz was used on urea and urine phantoms and normal human subjects imaged through the kidneys. The positive frequency was used as the control for each frequency pair. Results of frequency sweeps show an asymmetric MT effect peaking at approximately 100 Hz ( thick similar1 ppm) for urea, urine, and renal parenchyma. Varying differences (5%-25%) occurred with different human subjects. Few differences were observed from phantom water or subject muscle tissue. Chemical exchange is detectable in the kidney near 1 ppm at 1.5 T, attributable to urea. This technique was used to produce in vivo distribution maps of this metabolite in vivo. PMID- 11050644 TI - Age-related blood half-life of particulate contrast material: experimental results with a USPIO in rats. AB - It has been well established in the literature that phagocytic activity in animals and humans changes with age, but this phenomenon has not yet been investigated for particulate contrast agents. The present study was therefore performed to determine the effect of age on blood half-life of a superparamagnetic iron oxide blood pool contrast agent and on the velocity of its uptake in the liver and spleen of the rat by means of magnetic resonance (MR) imaging. A total of 18 rats (group A: 214-255 g, age: 45-50 days; group B: 432 563 g, age: 100-120 days) were imaged at 1.5 T using a 3D gradient-recalled echo (GRE) sequence (TR/TE 6.6/2.3 msec; alpha 25 degrees, frontal). Images were acquired before and every 3-5 minutes for up to 30 minutes after i.v. injection of 30 micromol Fe/kg of a citrate-coated superparamagnetic iron oxide-based contrast agent (VSOP-C43). Intravenous injection of VSOP-C43 resulted in a pronounced initial signal enhancement in vessels, which decreased with a half life of 8.4 +/- 0.9 minutes in group A and of 15.9 +/- 2. 4 minutes in group B (P < 0.01). The half-life of signal decrease was 11.6 +/- 2 minutes and 19.9 +/- 4.4 minutes in the liver (P < 0. 01) and 19.6 +/- 3.1 minutes and 26.7 +/- 5.2 minutes in the spleen (not significant). The results show that the age-related phagocytic activity has a significant effect on the circulation time and velocity of uptake of a particulate MR imaging contrast agent. This fact must be taken into consideration in both the preclinical and clinical development of particulate contrast material and in the clinical application of such agents. PMID- 11050646 TI - Estimation of magnetization transfer rates from PACE experiments with pulsed RF saturation. AB - A new imaging method has been developed for estimating the magnetization transfer rate (MTR) in a biologic two-pool system such as the brain tissue. The transfer rate is calculated from the ratio of the MTR to T(1sat), where T(1sat) is the apparent longitudinal relaxation time under complete saturation of the macromolecular pool. MTR and T(1sat) maps were obtained with a phase acquisition of composite echo (PACE) technique combined with pulsed radiofrequency (RF) saturation. The influences of RF saturation power and frequency offset on quantitative results were investigated with phantom and in vivo measurements. In white matter of seven healthy volunteers we found a mean transfer rate of 1.5 sec(-1), where the highest transfer rate was found in the genu of the corpus callosum (k(f) = 1. 9 sec(-1)). It could be shown that conditions near to complete saturation can also be reached under common restrictions by the specific absorption rate. PMID- 11050647 TI - Changes in baseline cerebral blood flow in humans do not influence regional cerebral blood flow response to photic stimulation. AB - The effect of changes in baseline regional cerebral blood flow (rCBF) values on the cerebral blood flow response during neuronal activation was studied with functional magnetic resonance imaging (fMRI). Using a breath-holding challenge as a hypercapnic stimulus, rCBF alterations during photic stimulation under normo- and hypercapnia were determined in nine volunteers. With breath-holding, baseline rCBF in areas corresponding to the visual cortex significantly increased from 54 +/- 5 ml/100 g/min to 85 +/- 9 ml/100 g/min (P < 0.001). Despite this significant change in baseline flow values, the rCBF increase during visual stimulation was very similar under normo- and hypercapnic conditions (28 +/- 8 ml/100 g/min versus 26 +/- 8 ml/100 g/min, respectively). This study supports the notion that within wide physiologic variations, task-induced cerebral blood flow changes are independent of baseline rCBF values. PMID- 11050648 TI - Pseudothrombosis in the portal venous system: a potential pitfall with gadolinium enhanced dynamic gradient-recalled echo imaging of the liver. AB - Gadolinium-enhanced dynamic magnetic resonance (MR) images in 90 patients were reviewed to assess the artifacts mimicking portal venous thrombosis. The incidence of definite signal-intensity decrease mimicking pathologic condition was higher (P < 0.01) in the right (8%) and left (9%) portal vein branches and portal trunk (6%) than in the splenic (0%) or superior mesenteric (1%) vein with equilibrium-phase images. Radiologists should remember that dynamic MR images occasionally show signal-intensity decrease mimicking portal venous thrombosis due to flow artifact. PMID- 11050649 TI - Bolus-chase peripheral 3D MRA using a dual-rate contrast media injection. AB - In this pilot study, using a standard 40 mL gadolinium (Gd) chelate contrast dose, dual-rate (first 20 mL at 0.5 mL/sec; remaining 20 mL at 1.5 mL/sec) and fixed-rate (entire 40 mL dose at either 0.7 mL/sec or 2.0 mL/sec) injection schemes for multistation, bolus-chase magnetic resonance angiography (MRA) were compared in normal volunteers. Signal-to-noise ratio, contrast-to-noise ratio, and physician preference were determined for nine arterial segments. At the terminal station (calf), the dual-rate contrast injection improved arterial signal and contrast compared with both fixed-rate injection schemes and improved subjective vessel appearance compared with the 2.0 mL/sec, but not the 0.7 mL/sec, fixed-rate scheme. PMID- 11050650 TI - Non-contrast-enhanced MR angiography using 3D ECG-synchronized half-Fourier fast spin echo. AB - A non-contrast-enhanced three-dimensional (3D) magnetic resonance angiography (MRA) technique, which acquires images in a reasonably short scanning time and requires no contrast agent, is developed. An electrocardiographically (ECG) synchronized 3D half-Fourier fast spin-echo (FSE) technique with an appropriate ECG delay time for every slice encoding in 3D terms was used to examine the thoracic and iliac regions in 16 healthy volunteers at both 0.5 and 1.5 T. Prior to each 3D fresh blood imaging (FBI) experiment, an ECG preparation (ECG-prep) scan was acquired, and an appropriate ECG triggering time was selected for 3D FBI acquisition to optimize visualization of the vessel of interest. In the thoracic and abdominal regions, good-quality 3D MRA images were obtained. Furthermore, the weighted subtraction of two images in different phases provides contrast enhancement between arteries and veins. PMID- 11050651 TI - A new fast and unsynchronized method for MRI of viscoelastic properties of soft tissues. AB - Quantitative measurement of mechanical properties of biologic tissues may have several applications for diagnosis or biomechanic modeling in sports medicine, traumatology, or computer-guided surgery. The magnetic resonance imaging (MRI) methods previously tested for these applications all required synchronization between MRI acquisition pulses and the mechanical stimulation. A new unsynchronized method operating with no prior knowledge of intensity, direction, and frequency of the mechanical waves is proposed. A specifically modified SPAMM (SPAtial Modulation of Magnetization) sequence has been used, operating on a 0.2 T MRI system. The experimental results obtained on test objects fit well with theoretical calculations. The new proposed method is very fast (a less than 5 second acquisition time) for routine clinical use. PMID- 11050652 TI - Noninvasive in vivo magnetic resonance imaging of injury-induced neointima formation in the carotid artery of the apolipoprotein-E null mouse. AB - Mice deficient in apolipoprotein-E (apoE) experience severe hypercholesterolemia, are prone to atherosclerosis, and recently have emerged as a powerful tool in the study of plaque formation. In this study, we developed magnetic resonance (MR) imaging methods to detect the progression of atherosclerosis noninvasively in a mouse model of arterial injury. Four 14-week-old apoE-deficient mice were imaged 5 weeks after beginning an atherogenic Western diet and 4 weeks after wire denudation injury of the left common carotid artery (LCCA). Information from several images was combined into high-information content images using methods previously developed. The image resolution was 47 x 47 x 750 microm(3). We acquired T1-, T2-, and proton density (PD)-weighted images (TR/TE 650/14, 2000/60, and 2000/14 msec, respectively). Each 8-bit image was placed in a separate color channel to produce a 24-bit color image (red = T1, green = PD, and blue = T2). The composite image created contrast between different tissue types that was superior to that of any single image and revealed significant luminal narrowing of the LCCA, but not the uninjured RCCA. MR images were compared with corresponding histopathology cross sections and luminal area measurements from each method correlated(r2= 0.61). Atherosclerotic luminal narrowing was successfully detected through MR imaging in a mouse model of arterial injury that is small, reproduces quickly, and lends itself to genetic analysis and manipulation. PMID- 11050653 TI - Elimination of magnetic field foldover artifacts in MR images. AB - Foldover artifacts arise when the same imaging frequency occurs both at a desired location within a slice and at another location within the sensitive region of the radiofrequency (RF) coil. Foldover artifacts can be caused by nonlinearity in the gradient system and by inhomogeneity in B(0). This study investigates an approach in which an extra RF receiver coil and a postprocessing method are used to identify and remove foldover artifacts. PMID- 11050655 TI - Typing of Enterococcus faecium by polymerase chain reaction and pulsed field gel electrophoresis. AB - Polymerase chain reaction (PCR) with JB1 or REP consensus oligonucleotides and pulsed field gel electrophoresis (PFGE) were used to study genomic DNA extracted from 31 strains of enterococci. Eleven ATCC strains, representative of 11 species of Enterococcus, were initially tested by JB1-PCR, revealing that Enterococcus malodoratus and Enterococcus hirae presented identical banding patterns. Eight Enterococcus faecium isolates from Stanford University and 12 from Sao Paulo Hospital were studied by JB1-PCR, REP-PCR (1/2)R and PFGE. Among the isolates from Stanford University, 5 genotypes were defined by JB1-PCR, 7 by REP-PCR (1/2)R and 4 by PFGE. Among the isolates from Sao Paulo Hospital, 9 genotypes were identified by JB1-PCR, 6 by REP-PCR and 5 by PFGE. The three methods identified identical genotypes, but there was not complete agreement among them. PMID- 11050654 TI - Effect of fatty acids on leukocyte function. AB - Fatty acids have various effects on immune and inflammatory responses, acting as intracellular and intercellular mediators. Polyunsaturated fatty acids (PUFAs) of the omega-3 family have overall suppressive effects, inhibiting lymphocyte proliferation, antibody and cytokine production, adhesion molecule expression, natural killer cell activity and triggering cell death. The omega-6 PUFAs have both inhibitory and stimulatory effects. The most studied of these is arachidonic acid that can be oxidized to eicosanoids, such as prostaglandins, leukotrienes and thromboxanes, all of which are potent mediators of inflammation. Nevertheless, it has been found that many of the effects of PUFA on immune and inflammatory responses are not dependent on eicosanoid generation. Fatty acids have also been found to modulate phagocytosis, reactive oxygen species production, cytokine production and leukocyte migration, also interfering with antigen presentation by macrophages. The importance of fatty acids in immune function has been corroborated by many clinical trials in which patients show improvement when submitted to fatty acid supplementation. Several mechanisms have been proposed to explain fatty acid modulation of immune response, such as changes in membrane fluidity and signal transduction pathways, regulation of gene transcription, protein acylation, and calcium release. In this review, evidence is presented to support the proposition that changes in cell metabolism also play an important role in the effect of fatty acids on leukocyte functioning, as fatty acids regulate glucose and glutamine metabolism and mitochondrial depolarization. PMID- 11050656 TI - Evidence for the expression of native Mycobacterium tuberculosis phospholipase C: recognition by immune sera and detection of promoter activity. AB - The genome of Mycobacterium tuberculosis H37Rv contains three contiguous genes (plc-a, plc-b and plc-c) which are similar to the Pseudomonas aeruginosa phospholipase C (PLC) genes. Expression of mycobacterial PLC-a and PLC-b in E. coli and M. smegmatis has been reported, whereas expression of the native proteins in M. tuberculosis H37Rv has not been demonstrated. The objective of the present study was to demonstrate that native PLC-a is expressed in M. tuberculosis H37Rv. Sera from mice immunized with recombinant PLC-a expressed in E. coli were used in immunoblots to evaluate PLC-a expression. The immune serum recognized a 49-kDa protein in immunoblots against M. tuberculosis extracts. No bands were visible in M. tuberculosis culture supernatants or extracts from M. avium, M. bovis and M. smegmatis. A 550-bp DNA fragment upstream of plc-a was cloned in the pJEM12 vector and the existence of a functional promoter was evaluated by detection of beta-galactosidase activity. beta-Galactosidase activity was detected in M. smegmatis transformed with recombinant pJEM12 grown in vitro and inside macrophages. The putative promoter was active both in vitro and in vivo, suggesting that expression is constitutive. In conclusion, expression of non-secreted native PLC-a was demonstrated in M. tuberculosis. PMID- 11050657 TI - Cloning and characterization of Echinococcus granulosus (Cestode) EgactI and EgactII actin gene promoters and their functional analysis in the NIH3T3 mouse cell line. AB - We report here for the first time the structure and function of a promoter from a cestode. The ability of DNA fragments respectively encompassing the 935-bp and 524-bp regions upstream from the ATG codon from the EgactI and EgactII actin genes of Echinococcus granulosus to promote transcription was studied in the NIH3T3 mouse cell line. The results of transfection assays showed that both regions have strong promoter activity in these cells. The fragments were tested in both orientations and the 524-bp fragment of EgactII presented a bidirectional promoter activity. Deletion analysis of EgactI and EgactII promoters indicated the presence of regulatory regions containing putative silencer elements. These results indicate that both EgactI and EgactII promoters are functional and that the preliminary functional evaluation of E. granulosus and possibly of other cestode promoters can be performed in heterologous cell lines. PMID- 11050658 TI - Two related thrombin-like enzymes present in Bothrops atrox venom. AB - This article describes the presence of two new forms of a thrombin-like enzyme, both with apparent molecular masses of 38 kDa, in Bothrops atrox venom. Both share the ability to cleave fibrinogen into fibrin and to digest casein. Both present identical K(m) on the substrate BApNA. Their N-terminal amino acid sequences are identical for 26 residues, sharing 80% homology with batroxobin and flavoxobin. Two groups of monoclonal antibodies (mAbs) raised against the purified enzyme forms recognized different epitopes of the putative corresponding enzymes present in B. atrox crude venom. On Western blotting analysis of B. atrox crude venom, mAbs 5DB2C8, 5AA10 and 5CF11, but not mAbs 6CC5 and 6AD2-G5, revealed two or more protein bands ranging from 25 to 38 kDa. By immunoprecipitation assays, the 6AD2-G5 mAb was able to precipitate protein bands of 36-38 kDa from B. atrox, B. leucurus, B. pradoi, B. moojeni, B. jararaca and B. neuwiedii crude venoms. Fibrinogen-clotting activity was inhibited when the same venom specimens were pre-incubated with mAb 6AD2-G5, except for B. jararaca and B. neuwiedii. PMID- 11050659 TI - Polymorphisms of the low-density lipoprotein receptor gene in Brazilian individuals with heterozygous familial hypercholesterolemia. AB - Familial hypercholesterolemia (FH) is a metabolic disorder inherited as an autosomal dominant trait characterized by an increased plasma low-density lipoprotein (LDL) level. The disease is caused by several different mutations in the LDL receptor gene. Although early identification of individuals carrying the defective gene could be useful in reducing the risk of atherosclerosis and myocardial infarction, the techniques available for determining the number of the functional LDL receptor molecules are difficult to carry out and expensive. Polymorphisms associated with this gene may be used for unequivocal diagnosis of FH in several populations. The aim of our study was to evaluate the genotype distribution and relative allele frequencies of three polymorphisms of the LDL receptor gene, HincII(1773) (exon 12), AvaII (exon 13) and PvuII (intron 15), in 50 unrelated Brazilian individuals with a diagnosis of heterozygous FH and in 130 normolipidemic controls. Genomic DNA was extracted from blood leukocytes by a modified salting-out method. The polymorphisms were detected by PCR-RFLP. The FH subjects showed a higher frequency of A+A+ (AvaII), H+H+ (HincII(1773)) and P1P1 (PvuII) homozygous genotypes when compared to the control group (P<0.05). In addition, FH probands presented a high frequency of A+ (0.58), H+ (0.61) and P1 (0.78) alleles when compared to normolipidemic individuals (0.45, 0.45 and 0.64, respectively). The strong association observed between these alleles and FH suggests that AvaII, HincII(1773) and PvuII polymorphisms could be useful to monitor the inheritance of FH in Brazilian families. PMID- 11050660 TI - Nutritional status of hemodialysis patients with secondary hyperparathyroidism. AB - The repercussions of secondary hyperparathyroidism on the nutritional status of chronic renal failure patients have not been well established. Therefore, the aim of this study was to compare the nutritional indices of hemodialysis patients with and without secondary hyperparathyroidism. Sixteen hemodialysis patients with serum parathyroid hormone (PTH) levels higher than 420 pg/ml (hyperparathyroidism group) were matched for gender, age and length of dialysis treatment to 16 patients with serum PTH between 64 and 290 pg/ml (control group). The following parameters were assessed: anthropometric indices (body mass index, skinfold thickness, midarm muscle circumference and body fat), 4-day food diaries, protein catabolic rate, biochemical indices (blood urea nitrogen, serum creatinine, albumin, ionized calcium, inorganic phosphorus, serum alkaline phosphatase, PTH, pH and HCO(3)) and dialysis efficiency. We did not observe differences in the anthropometric indices between the two groups. Only calcium intake was significantly different between groups (307.9 mg/day for the hyperparathyroidism group vs 475.8 mg/day for the control group). Protein catabolic rate tended to be higher in the hyperparathyroidism group compared to the control group (1.3 vs 0.9 g kg(-1) day(-1); P = 0.08). Except for blood urea nitrogen (86.4 vs 75.7 mg/dl), alkaline phosphatase (175 vs 65 U/l) and PTH (898 vs 155 pg/ml), no other differences were found between groups in the biochemical indices studied. PTH was directly correlated with protein catabolic rate (r = 0.61; P<0.05) and length of dialysis (r = 0.53; P<0.05) only in the hyperparathyroidism group. Considering the indices used, we could not demonstrate the deleterious effect of high PTH levels on the nutritional status of hemodialysis patients. Indirect evidence, however, suggests an action of PTH on protein metabolism. PMID- 11050661 TI - Increased fetal hemoglobin levels in patients infected with human immunodeficiency virus (HIV1/2). AB - Fetal hemoglobin was measured in HIV1/2 patients under treatment with combined therapy (zidovudine and a protease inhibitor). A total of 143 patients and 103 normal individuals were investigated by the quantitative method of Betke and the semi-quantitative acid elution method of Kleihauer. In the normal person, hemoglobin F makes up less than 1% and an increase higher than 1.5% was observed in 21.4% of HIV patients by the method of Betke and in 24.8% of HIV-infected patients by the method of Kleihauer. The quantitative biochemical method of Betke showed that the populations were significantly different (two-tailed Mann-Whitney test). The reason for this hemoglobin F increase might be ascribed to the effect of zidovudine or to direct viral action on gamma chain expression. The finding of a higher F cell frequency indicated by the method of Kleihauer rather suggests that there is an increased F cell clone proliferation rather than an increase in hemoglobin F level in every cell. PMID- 11050662 TI - Hyperventilation in panic disorder patients and healthy first-degree relatives. AB - Our aim was to observe the induction of panic attacks by a hyperventilation challenge test in panic disorder patients (DSM-IV) and their healthy first-degree relatives. We randomly selected 25 panic disorder patients, 31 healthy first degree relatives of probands with panic disorder and 26 normal volunteers with no family history of panic disorder. All patients had no psychotropic drugs for at least one week. They were induced to hyperventilate (30 breaths/min) for 4 min and anxiety scales were applied before and after the test. A total of 44.0% (N = 11) panic disorder patients, 16.1% (N = 5) of first-degree relatives and 11.5% (N = 3) of control subjects had a panic attack after hyperventilating (chi(2) = 8.93, d.f. = 2, P = 0.011). In this challenge test the panic disorder patients were more sensitive to hyperventilation than first-degree relatives and normal volunteers. Although the hyperventilation test has a low sensitivity, our data suggest that there is no association between a family history of panic disorder and hyperreactivity to an acute hyperventilation challenge test. Perhaps cognitive variables should be considered to play a specific role in this association since symptoms of a panic attack and acute hyperventilation overlap. PMID- 11050663 TI - Investigation of single-strand conformational polymorphism of the TP53 gene in women with a family history of breast cancer. AB - Breast cancer in families with germ line mutations in the TP53 gene has been described in the medical literature. Mutation screening for susceptibility genes should allow effective prophylactic and preventive measures. Using single-strand conformational polymorphism, we screened for mutations in exons 5, 6, 7 and 8 of gene TP53 in the peripheral blood of 8 young non-affected members (17 to 36 years old) of families with a history of breast cancer. Studies of this type on young patients (mean age, 25 years) are very rare in the literature. The identification of these mutations would contribute to genetic counseling of members of families with predisposition to breast cancer. The results obtained did not show any polymorphism indicating mutation. In our sample, the familial tumorigenesis is probably related to other gene etiologies. PMID- 11050664 TI - Hepatic capillariasis in rats: a new model for testing antifibrotic drugs. AB - Rats infected with the helminth Capillaria hepatica regularly develop septal hepatic fibrosis that may progress to cirrhosis in a relatively short time. Because of such characteristics, this experimental model was selected for testing drugs exhibiting antifibrosis potential, such as pentoxifylline, gadolinium chloride and vitamin A. Hepatic fibrosis was qualitatively and quantitatively evaluated in liver samples obtained by partial hepatectomy and at autopsy. The material was submitted to histological, biochemical and morphometric methods. A statistically significant reduction of fibrosis was obtained with pentoxifylline when administered intraperitoneally rather than intravenously. Gadolinium chloride showed moderate activity when administered prophylactically (before fibrosis had started), but showed a poor effect when fibrosis was well advanced. No modification of fibrosis was seen after vitamin A administration. Hydroxyproline content was correlated with morphometric measurements. The model appears to be adequate, since few animals die of the infection, fibrosis develops regularly in all animals, and the effects of different antifibrotic drugs and administration protocols can be easily detected. PMID- 11050665 TI - Pancreatic nitric oxide and oxygen free radicals in the early stages of streptozotocin-induced diabetes mellitus in the rat. AB - The objective of the present study was to explore the regulatory mechanisms of free radicals during streptozotocin (STZ)-induced pancreatic damage, which may involve nitric oxide (NO) production as a modulator of cellular oxidative stress. Removal of oxygen species by incubating pancreatic tissues in the presence of polyethylene glycol-conjugated superoxide dismutase (PEG-SOD) (1 U/ml) produced a decrease in nitrite levels (42%) and NO synthase (NOS) activity (50%) in diabetic but not in control samples. When NO production was blocked by N(G)-monomethyl-L arginine (L-NMMA) (600 microM), SOD activity increased (15.21 +/- 1.23 vs 24.40 +/- 2.01 U/mg dry weight). The increase was abolished when the NO donor, spermine nonoate, was added to the incubating medium (13.2 +/- 1.32). Lipid peroxidation was lower in diabetic tissues when PEG-SOD was added (0.40 +/- 0.02 vs 0.20 +/- 0.03 nmol/mg protein), and when L-NMMA blocked NOS activity in the incubating medium (0.28 +/- 0.05); spermine nonoate (100 microM) abolished the decrease in lipoperoxide level (0.70 +/- 0.02). We conclude that removal of oxygen species produces a decrease in pancreatic NO and NOS levels in STZ-treated rats. Moreover, inhibition of NOS activity produces an increase in SOD activity and a decrease in lipoperoxidation in diabetic pancreatic tissues. Oxidative stress and NO pathway are related and seem to modulate each other in acute STZ-induced diabetic pancreas in the rat. PMID- 11050666 TI - Interaction between repeated restraint stress and concomitant midazolam administration on sweet food ingestion in rats. AB - Emotional changes can influence feeding behavior. Previous studies have shown that chronically stressed animals present increased ingestion of sweet food, an effect reversed by a single dose of diazepam administered before testing the animals. The aim of the present study was to evaluate the response of animals chronically treated with midazolam and/or submitted to repeated restraint stress upon the ingestion of sweet food. Male adult Wistar rats were divided into two groups: controls and exposed to restraint 1 h/day, 5 days/week for 40 days. Both groups were subdivided into two other groups treated or not with midazolam (0.06 mg/ml in their drinking water during the 40-day treatment). The animals were placed in a lighted area in the presence of 10 pellets of sweet food (Froot loops). The number of ingested pellets was measured during a period of 3 min, in the presence or absence of fasting. The group chronically treated with midazolam alone presented increased ingestion when compared to control animals (control group: 2.0 +/- 0.44 pellets and midazolam group: 3.60 +/- 0.57 pellets). The group submitted to restraint stress presented an increased ingestion compared to controls (control group: 2.0 +/- 0.44 pellets and stressed group: 4.18 +/- 0.58 pellets). Chronically administered midazolam reduced the ingestion in stressed animals (stressed/water group: 4.18 +/- 0.58 pellets; stressed/midazolam group: 3.2 +/- 0.49 pellets). Thus, repeated stress increases appetite for sweet food independently of hunger and chronic administration of midazolam can decrease this behavioral effect. PMID- 11050667 TI - Molluscicidal activity of Punica granatum bark and Canna indica root. AB - The molluscicidal activity of Punica granatum Linn. (Punicaceae) and Canna indica Linn. (Cannaceae) against the snail Lymnaea acuminata was studied. The molluscicidal activity of P. granatum bark and C. indica root was found to be both time and dose dependent. The toxicity of P. granatum bark was more pronounced than that of C. indica. The 24 h LC(50) of the column-purified root of C. indica was 6.54 mg/l whereas that of the column-purified bark of P. granatum was 4.39 mg/l. The ethanol extract of P. granatum (24 h LC(50): 22.42 mg/l) was more effective than the ethanol extract of C. indica (24 h LC(50): 55.65 mg/l) in killing the test animals. P. granatum and C. indica may be used as potent molluscicides since the concentrations used to kill the snails were not toxic for the fish Colisa fasciatus, which shares the same habitat with the snail L. acuminata. PMID- 11050668 TI - Abnormalities of glucose metabolism in spontaneously hypertensive rats. AB - Abnormalities in glucose metabolism and insulin action are frequently detected in patients with essential hypertension. Spontaneously hypertensive rats (SHR) have been used as an experimental model to understand this pathological condition. The objective of the present study was to assess glucose metabolism and insulin action in SHR and Wistar rats under fed and fasting conditions. Peripheral glucose utilization was estimated by kinetic studies with [6-(3)H]-glucose and gluconeogenetic activity was measured during continuous [(14)C]-bicarbonate infusion. Plasma glucose levels were higher in the SHR group. Plasma insulin levels in the fed state were higher in the SHR group (99.8 +/- 6.5 microM) than in the control group (70.4 +/- 3.6 microM). Muscle glycogen content was reduced in SHR compared to control under the various experimental conditions. Peripheral glucose utilization was slightly lower in the SHR group in the fed state (8.72 +/ 0.55 vs 9.52 +/- 0.80 mg kg(-1) min(-1) in controls). Serum free fatty acid levels, hepatic glycogen levels, hepatic phosphoenolpyruvate carboxykinase activity and gluconeogenetic activity were similar in the two groups. The presence of hyperglycemia and hyperinsulinemia and the slightly reduced peripheral glucose utilization suggest the presence of resistance to the action of insulin in peripheral tissues of SHR. Hepatic gluconeogenesis does not seem to contribute to the metabolic alterations detected in these animals. PMID- 11050669 TI - Oxidative stress in the latissimus dorsi muscle of diabetic rats. AB - The purpose of the present study was to investigate the effects of experimental diabetes on the oxidant and antioxidant status of latissimus dorsi (LD) muscles of male Wistar rats (220 +/- 5 g, N = 11). Short-term (5 days) diabetes was induced by a single injection of streptozotocin (STZ, 50 mg/kg, iv; glycemia >300 mg/dl). LD muscle of STZ-diabetic rats presented higher levels of thiobarbituric acid reactive substances (TBARS) and chemiluminescence (0.36 +/- 0.02 nmol/mg protein and 14706 +/- 1581 cps/mg protein) than LD muscle of normal rats (0.23 +/ 0.04 nmol/mg protein and 7389 +/- 1355 cps/mg protein). Diabetes induced a 92% increase in catalase and a 27% increase in glutathione S-transferase activities in LD muscle. Glutathione peroxidase activity was reduced (58%) in STZ-diabetic rats and superoxide dismutase activity was similar in LD muscle of both groups. A positive correlation was obtained between catalase activity and the oxidative stress of LD, as evaluated in terms of TBARS (r = 0.78) and by chemiluminescence (r = 0.89). Catalase activity also correlated inversely with glutathione peroxidase activity (r = 0.79). These data suggest that an increased oxidative stress in LD muscle of diabetic rats may be related to skeletal muscle myopathy. PMID- 11050670 TI - Brain ischemia alters platelet ATP diphosphohydrolase and 5'-nucleotidase activities in naive and preconditioned rats. AB - The effects of transient forebrain ischemia, reperfusion and ischemic preconditioning on rat blood platelet ATP diphosphohydrolase and 5'-nucleotidase activities were evaluated. Adult Wistar rats were submitted to 2 or 10 min of single ischemic episodes, or to 10 min of ischemia 1 day after a 2-min ischemic episode (ischemic preconditioning) by the four-vessel occlusion method. Rats submitted to single ischemic insults were reperfused for 60 min and for 1, 2, 5, 10 and 30 days after ischemia; preconditioned rats were reperfused for 60 min 1 and 2 days after the long ischemic episode. Brain ischemia (2 or 10 min) inhibited ATP and ADP hydrolysis by platelet ATP diphosphohydrolase. On the other hand, AMP hydrolysis by 5'-nucleotidase was increased after 2, but not 10, min of ischemia. Ischemic preconditioning followed by 10 min of ischemia caused activation of both enzymes. Variable periods of reperfusion distinctly affected each experimental group. Enzyme activities returned to control levels in the 2 min group. However, the decrease in ATP diphosphohydrolase activity was maintained up to 30 days of reperfusion after 10-min ischemia. 5'-Nucleotidase activity was decreased 60 min and 1 day following 10-min ischemia; interestingly, enzymatic activity was increased after 2 and 5 days of reperfusion, and returned to control levels after 10 days. Ischemic preconditioning cancelled the effects of 10-min ischemia on the enzymatic activities. These results indicate that brain ischemia and ischemic preconditioning induce peripheral effects on ecto-enzymes from rat platelets involved in nucleotide metabolism. Thus, ATP, ADP and AMP degradation and probably the generation of adenosine in the circulation may be altered, leading to regulation of microthrombus formation since ADP aggregates platelets and adenosine is an inhibitor of platelet aggregation. PMID- 11050671 TI - Blockade of NK-1 receptors in the lateral commissural nucleus tractus solitarii of awake rats had no effect on the cardiovascular responses to chemoreflex activation. AB - The neurotransmission of the chemoreflex in the nucleus tractus solitarii (NTS), particularly of the sympatho-excitatory component, is not completely understood. There is evidence that substance P may play a role in the neurotransmission of the chemoreflex in the NTS. Microinjection of substance P (50 pmol/50 nl, N = 12, and 5 nmol/50 nl, N = 8) into the commissural NTS of unanesthetized rats produced a significant increase in mean arterial pressure (101 +/- 1 vs 108 +/- 2 and 107 +/- 3 vs 115 +/- 4 mmHg, respectively) and no significant changes in heart rate (328 +/- 11 vs 347 +/- 15 and 332 +/- 7 vs 349 +/- 13 bpm, respectively) 2 min after microinjection. Previous treatment with WIN, an NK-1 receptor antagonist (2.5 nmol/50 nl), microinjected into the NTS of a specific group of rats, blocked the pressor (11 +/- 5 vs 1 +/- 2 mmHg) and tachycardic (31 +/- 6 vs 4 +/- 3 bpm) responses to substance P (50 pmol/50 nl, N = 5) observed 10 min after microinjection. Bilateral microinjection of WIN into the lateral commissural NTS (N = 8) had no significant effect on the pressor (50 +/- 4 vs 42 +/- 6 mmHg) or bradycardic (-230 +/- 16 vs -220 +/- 36 bpm) responses to chemoreflex activation with potassium cyanide (iv). These data indicate that the activation of NK-1 receptors by substance P in the NTS produces an increase in baseline mean arterial pressure and heart rate. However, the data obtained with WIN suggest that substance P and NK-1 receptors do not play a major role in the neurotransmission of the chemoreflex in the lateral commissural NTS. PMID- 11050672 TI - Cycle modulation of insulin-like growth factor-binding protein 1 in human endometrium. AB - Endometrium is one of the fastest growing human tissues. Sex hormones, estrogen and progesterone, in interaction with several growth factors, control its growth and differentiation. Insulin-like growth factor 1 (IGF-1) interacts with cell surface receptors and also with specific soluble binding proteins. IGF-binding proteins (IGF-BP) have been shown to modulate IGF-1 action. Of six known isoforms, IGF-BP-1 has been characterized as a marker produced by endometrial stromal cells in the late secretory phase and in the decidua. In the current study, IGF-1-BP concentration and affinity in the proliferative and secretory phase of the menstrual cycle were measured. Endometrial samples were from patients of reproductive age with regular menstrual cycles and taking no steroid hormones. Cytosolic fractions were prepared and binding of (125)I-labeled IGF-1 performed. Cross-linking reaction products were analyzed by SDS-polyacrylamide gel electrophoresis (7.5%) followed by autoradiography. (125)I-IGF-1 affinity to cytosolic proteins was not statistically different between the proliferative and secretory endometrium. An approximately 35-kDa binding protein was identified when (125)I-IGF-1 was cross-linked to cytosol proteins. Secretory endometrium had significantly more IGF-1-BP when compared to proliferative endometrium. The specificity of the cross-linking process was evaluated by the addition of 100 nM unlabeled IGF-1 or insulin. Unlabeled IGF-1 totally abolished the radioactivity from the band, indicating specific binding. Insulin had no apparent effect on the intensity of the labeled band. These results suggest that IGF-BP could modulate the action of IGF-1 throughout the menstrual cycle. It would be interesting to study this binding protein in other pathologic conditions of the endometrium such as adenocarcinomas and hyperplasia. PMID- 11050682 TI - Bile duct ligation and oxidative stress in the rat: effects in liver and kidney. AB - In the liver, seven days of bile duct ligation (BDL) decreases the cytochrome P 450 content and the UDP-glucuronyl transferase activity. Also, a decrease in the water soluble antioxidant mechanism reflected in the activities of the enzymes superoxide dismutase (SOD), catalase and the glutathione peroxidase (GTPx) was found in the liver but not in the kidney. Despite an increase in the amount of the GSH in the liver, increased lipid peroxidation is produced in the BDL rats, as indicated by the levels of malondialdehyde (MDA). The kidney responded in a different way to cholestasis, decreasing only the UDP-glucuronyl transferase activity and increasing the levels of GSH and MDA. In the red blood cells the activity of the antioxidant enzymes SOD, GTPx and catalase and the content of GSH were not modulated by cholestasis. In conclusion, disturbance of the oxidant antioxidant balance might be responsible for cholestatic liver injury and impaired renal function in BDL rats. PMID- 11050683 TI - Metallothioneins in liver of Rutilus rutilus exposed to Cu2+. Analysis by metal summation, SH determination and spectrofluorimetry. AB - Metallothioneins (MTs) have important roles in the homeostasis of essential metals and in the detoxication of heavy metals. They also represent a potential indicator of aquatic contamination by metals. Routine methods are needed for MTs quantification in ecotoxicological studies. This paper investigates the possibility to use the spectrofluorescent properties of Cu-MTs for MTs quantification. Cu displacement of metals coordinated to MTs and spectrofluorimetric determination of the obtained Cu-MTs was tested with commercial MTs and Cu2+-induced MTs in roach liver (Rutilus rutilus). Results of this original and simple spectrofluorimetric quantification of MTs presented a good correlation with data obtained with SH quantification, but not with metal summation evaluation of MTs (analysis of Zn, Cu and Cd coordinated to MTs). The three methods showed an clear induction of MTs in roach liver after 7 days of Cu2+ exposure. After 14 days of contamination, a reduction of hepatic MTs content was observable and not correlated to liver recovery. Results show that this low cost spectrofluorimetric method is useful to quantify MTs. PMID- 11050684 TI - Involvement of angiotensin II and endothelin-1 in the development of submandibular gland hypertrophy in response to isoproterenol in rats. AB - We investigated whether angiotensin II (Ang II) and endothelin-1(ET-1) are involved in submandibular hypertrophy in response to repeated treatment with isoproterenol (ISO) in rats. The immunoreactive Ang II (IRAng II) and immunoreactive ET-1 (IRET-1) contents of ISO-induced hypertrophy were significantly higher than those of control glands. Treatment of isolated gland tissues with ISO (1 microM) or dobutamine (1 microM) caused significant increases in the IRAng II and IRET- 1 contents of the glands compared with controls. These increases were suppressed by pretreatment with enalapril (3 microM) or captopril (3 microM). Treatment with Ang II (10 microM) also caused an increase in IRET-1 content. Our findings suggest that Ang II and ET-1 are involved in the submandibular gland hypertrophy that develops in rats repeatedly treated with ISO, and that these biologically active peptides may act as growth factors. They also imply that the tissue renin-angiotensin system and Ang II specific receptors are present in the submandibular glands. PMID- 11050685 TI - Vasopressin and A1 noradrenaline turnover during food or water deprivation in the rat. AB - In the present study, we have examined in Wistar rats the effects of food or water deprivation of 3 days on the hypophyso-adrenal axis, vasopressinergic system and activity of A1 noradrenergic brain stem cell group, which is involved in the control of the hypothalamic neuro-endocrine activity. Levels of adrenocorticotropic hormone (ACTH) and vasopressin (AVP) were determined by radio immunoassay, and corticosterone level was determined by fluorimetric method. Plasma levels of ACTH and corticosterone were greatly increased in both groups of rats. In water-deprived rats, plasma AVP (13.83 +/- 1.63 vs. 3.03 +/- 0.23 pg/ml) and osmolality levels were significantly elevated with a marked decrease of AVP hypophysis content (272 +/- 65 vs. 1098 +/- 75 ng/mg protein), but not in food deprived rats in which osmolality did not change and AVP remained stocked (2082 +/- 216 ng/mg protein) in the hypophysis without release in the plasma (1.11 +/- 0.23 pg/ml). These observations indicated that both food-deprivation and water deprivation stimulated the pituitary adrenal axis thereby suggesting a stress state. AVP production is stimulated both by fluid and food restriction but is secreted with differential effects: during food restriction AVP secretion is limited to supporting the hypothalamic pituitary-adrenal system. PMID- 11050686 TI - Effect of 2-amino-4-phosphonobutyrate on the OFF responses of frog retinal ganglion cells and local ERG after glycinergic blockade. AB - Perfusion with the ON channel blocker 2-amino-4-phosphonobutyrate (APB) of dark adapted frog eyecups not only abolished the ganglion cells' (GC) ON responses and the ERG b-wave, but markedly potentiated the OFF responses of ON-OFF and phasic OFF-GCs and the d-wave amplitude of simultaneously recorded local ERG. Glycinergic blockade by strychnine prevented this potentiating effect in 31 out of 69 GCs, but did not change it at all in the other cells. At the same time the d-wave potentiation was preserved during the glycinergic blockade in all eyecups. The results indicate that glycinergic transmission is involved in the inhibition exerted from ON upon OFF channel in some but not all frog retinal GCs. PMID- 11050687 TI - Levels of progesterone, testosterone, and estradiol, and androstenedione metabolism in the gonads of Lytechinus variegatus (Echinodermata:echinoidea). AB - Levels of progesterone (P4), testosterone (T) and estradiol (E2) indicated significant variation among individual echinoids during the annual cycle, reflecting generally the variation in gamete development that can be observed among individuals. Testosterone and E2 levels in both the ovaries and testes were higher during the period of gonadal growth. Levels of all steroids were greatly reduced compared to those levels reported for asteroids. Differences in the levels of P4, T, and estrogens between asteroids and Lytechinus variegatus may be related to differences in gonad morphology and nutrient storage capacity between asteroids and echinoids. It was hypothesized that the low levels of steroids detected in L. variegatus reflect paracrine-like mechanisms in cell signaling as compared to endocrine-like mechanisms proposed to be involved in regulating gonad function in asteroids. Both the ovaries and testes of L. variegatus had the capacity to synthesize T and a variety of 5alpha-reduced androgens including 5alpha-androstane-3beta,17beta-diol and 5alpha-androstane-3alpha,17beta-diol (5alpha-adiols) from androstenedione (AD) in 8 h. Estrogen synthesis was not detected. The sex-specific pattern of accumulation of 5alpha-adiols in the ovaries and testes suggests that the 5alpha-adiols may affect processes related to reproduction in L. variegatus. PMID- 11050688 TI - Distribution of [3H]cholesterol-labelled liposomes with or without praziquantel in mice infected with Mesocestoides corti (Cestoda) tetrathyridia. AB - The anthelmintic drug praziquantel (PZQ) has a short half-life in the circulation, necessitating repeated daily administration of PZQ for the therapy of larval stages of cestodes. The effect of incorporation of PZQ into multilamellar liposomes on their biodistribution in Mesocestoides corti (syn. M. vogae) infected mice has been examined using [3H]cholesterol as a liposomal marker. Incorporation of PZQ significantly increased the average size of liposomes with 70.3% of [3H]lip.PZQ particles up to 1.9 microm, whereas higher portion of [3H]liposomes (66.3% of total) were of smaller (up to 1.3 microm). Both liposome preparations were given intraperitoneally to avoid rapid sequestration in the liver. There were significant differences between [3H]liposomes and [3H]lip.PZQ-associated radioactivity in peritoneal adherent cells, liver- and peritoneal larvae, liver, spleen and lymph nodes within 16 days of examination. The highest uptake (about 2-fold more [3H]lip.PZQ than [3H]liposomes from the total dose) was found in peritoneal cells on day 1 post therapy (p.t.) followed by a rapid decline. The kinetic of decline in these cells recovered on day 1 p.t. was studied also in vitro. Disappearance of the marker due to the breakdown of liposomes and efflux of lipids and PZQ from cells was slower for [3H]lip.PZQ in comparison with drug-free liposomes and was not completed after 4 days-incubation. Significantly increased levels of radioactivity, more in [3H]liposomes treated groups, were recorded in the liver- and peritoneal larvae between days 8-16 p.t. indicating re-utilization of cholesterol by the larvae. The data suggest that incorporation of PZQ into liposomes contributes to the enlargement of liposome average size and slows down their degradation in phagocytosing cells. In this respect, these cells could serve as the secondary circulating depots for PZQ releasing it slowly to the circulation. PMID- 11050689 TI - Interspecies differences in P-glycoprotein mediated activity of multixenobiotic resistance mechanism in several marine and freshwater invertebrates. AB - The presence and function of the P-glycoprotein mediated multixenobiotic resistance (MXR) mechanism was demonstrated in numerous aquatic organisms. The aim of this study was to investigate whether in aquatic organisms exists the inherent, species-specific basal level of MXR activity. Here the results of the direct comparison of the basal (noninduced) level of MXR activity measured in several marine (Mytilus galloprovincialis, Monodonta turbinata, Patella lusitanica) and freshwater (Dreissena polymorpha, Viviparus viviparus, Anodonta cygnea) molluscs species are presented. The primary criterion for the assessment and quantification of the basal level of MXR activity was the ratio (R) between the accumulation or efflux of the fluorescent model MXR substrates (rhodamine B or rhodamine 123) in or from the gills, measured with and in the absence of model MXR inhibitors verapamil or cyclosporin A. Significantly different levels of MXR activity were found in the species investigated. These levels generally show a relatively good correlation with the level of pollution present in their natural habitats. Considering these results a conclusion was reached that in aquatic organisms indeed exist the different inherent, species-specific levels of MXR activity. The identified levels might be, at least partly, responsible either for the resistance to, or for the sensitivity of a particular species to organic pollution. PMID- 11050690 TI - Carbohydrate metabolism in temporal and persistent hypoglycemic chickens induced by insulin infusion. AB - In order to elucidate the regulatory mechanism of blood glucose concentrations specific to chickens, carbohydrate metabolism in the liver, muscle and kidney and metabolite concentrations in the blood were investigated in chickens with acute and persistent hypoglycemia. Acute and persistent hypoglycemia were experimentally induced by a single injection of insulin (8 U/kg BW) or by continuous infusion of insulin (22.5 U/kg BW/day) for 4 days. Non-esterified fatty acid (NEFA) concentration in plasma and D-3-hydroxybutyrate (3HB) concentrations in liver and muscle increased in the acute hypoglycemia. Plasma NEFA concentration and 3HB concentration in the blood and liver were not changed at day 3 of persistent hypoglycemia, while 3HB concentration in the muscle was decreased. Phosphofructokinase (PFK) activity in the liver tended to increase but PFK and pyruvate kinase (PK) activities were unchanged in acute hypoglycemia. In persistent hypoglycemia, increase of hepatic PFK activity at day 1 in which it was reversed at day 3, and a small increase of muscle PK activity were observed, while PK and phosphoenolpyruvate carboxykinase (PEPCK) activities in the liver and kidney were not significantly changed. These results show that in the persistent hypoglycemic chickens, hepatic glycolysis transiently increases, which is followed by a small decrease, while glycolysis in muscles and gluconeogenesis in the liver and kidney are not significantly changed. PMID- 11050691 TI - Effects of allyl mercaptan and various allium-derived compounds on cholesterol synthesis and secretion in Hep-G2 cells. AB - The present study was undertaken to compare the effects of allyl mercaptan (AM), a major metabolite of garlic, with several garlic constituents and extracts on cytotoxicity, cholesterol synthesis and its secretion in Hep-G2 cells. The cells were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), and treated with 5, 25, 50, 125, 250 and 500 microg/ml of AM, diallyl disulfide (DD), diallyl trisulfide (DT), steam-distilled garlic oil (SD) or vinyl-dithiin oil of garlic (VD) for 4 h. At concentrations up to 50 microg/ml, no significant cytotoxic effect was found in any group, but at concentrations above 250 microg/ml, the cell viability decreased drastically in all groups compared to the control. The treatment of cells with 25 microg/ml (non cytotoxic concentration) of AM, DD, DT, SD for 4 h significantly inhibited [3H]acetate incorporation into cholesterol compared to that of the control (P < 0.05). The secretion of cholesterol into the medium was also significantly decreased in all groups except for VD. The treatment of cells with those allium constituents had no effect on either [3H]acetate incorporation into fatty acids or [3H]glycerol incorporation into triglyceride or phospholipid. PMID- 11050692 TI - Circaannual changes in antioxidants and oxidative stress in the heart and liver in rats. AB - Reactive oxygen species are formed in physiological and pathological conditions in mammalian tissues. Because of their high reactivity, they may interact with biomolecules, inducing oxidative injury. Increases in lipid peroxidation can result in oxidative damage to cellular membranes. Protection against oxidative damage is provided by enzymatic and non-enzymatic antioxidant defenses. Antioxidant enzyme activities and lipid peroxidation, as an index of oxidative stress injury, were evaluated in different seasons over one year in the heart and liver of rats, maintained on a 12 h light and dark cycle. Glutathione peroxidase and catalase activities, in both tissues, were maximal in the summer season. Lipid peroxidation in the heart was maximal in the spring as compared to the other seasons and it did not vary in the liver during the year. These findings suggest that any study of antioxidants or oxidative stress must take into account such seasonal variations for a more precise analysis of changes due to any pathological condition. PMID- 11050693 TI - Bioavailability and pharmacokinetic profile of glyceryl trinitrate and of glyceryl dinitrates during application of a new glyceryl trinitrate transdermal patch. AB - The pharmacokinetics and bioavailability of glyceryl trinitrate (GTN, CAS 55-63 0) and of its main metabolites, i.e. 1,2-glyceryl dinitrate (1,2-GDN, CAS 621-65 8) and 1,3-glyceryl dinitrate (1,3-GDN, CAS 623-87-0), were compared during a single 24-h application of a new GTN transdermal patch (Epinitril 10, hereinafter called EPI-10) or a reference patch (hereinafter called ND-10) releasing 10 mg GTN in 24 h. The study was an open, randomized balanced cross-over study on 24 healthy male volunteers to whom the patches were applied to the antero-lateral part of the thorax in two periods separated by a 3-day wash-out. Blood samples were collected before administration, during the 24-h patch application and at 0.5, 2 and 3 h after patch removal. Assayed in plasma were GTN, 1,2-GDN and 1,3 GDN using validated GC/MS methods with stable isotope-labeled internal standards (15N3-GTN, 15N2-1,2-GDN, and 15N2-1,3-GDN). The ratios of the AUCs of GTN, 1,2 GDN and 1,3-GDN measured during application of EPI-10 or of ND-10 were within the 0.85-1.25 limits required to assess equivalence of the extent of bioavailability. The ratios of the Cmax were within said limits for the signal metabolite 1,2-GDN and only slightly below (0.78-1.16) for the parent GTN. EPI-10 can therefore be considered equivalent to ND-10 also with regard to the rate of bioavailability. Under both patches GTN reached steady-state levels after 3-6 h of patch application and remained on sustained levels during the whole 24-h application. The plasma levels of 1,2-GDN were about 6 times higher than those of GTN. The plasma levels of 1,3-GDN were similar to those of GTN. Upon removal of the patches the concentrations of the three nitrates fell to negligible values within 3 h. Both patches were well tolerated at the application site. For its small size, thinness and transparency, EPI-10 is very patient friendly, a quality that improves compliance with the therapeutic regimen. PMID- 11050694 TI - Plasma levels of glyceryl trinitrate and dinitrates during application of three strengths of a new glyceryl trinitrate transdermal patch. AB - The pharmacokinetic characteristics and the bioavailability of glyceryl trinitrate (GTN, CAS 55-63-0) and of its main metabolities 1,2-glyceryl dinitrate (1,2-GDN, CAS 621-65-8) and 1,3-glyceryl dinitrate (1,3-GDN, CAS 623-87-0) during a single 24-h application of three strengths of a newly developed GTN transdermal patch (Epinitril) were investigated. The three strengths coded in this paper EPI 5, EPI-10 and EPI-15 have a nominal release rate of GTN of 5, 10 and 15 mg, respectively, in 24 h. The study was an open, randomized balanced cross-over study on 18 healthy male volunteers, to whom the patches were applied for 24 h to the antero-lateral part of the thorax in three periods, separated by an at least 3-day wash-out. Blood samples were collected before administration, during the 24 h patch application and 1, 3 and 6 h after patch removal. Assayed in plasma were GTN, 1,2-GDN and 1,3-GDN using validated GC/MS methods with stable isotope labeled internal standards (15N3-GTN, 15N2-1,2-GDN, and 15N2-1,3-GDN). GTN and its two metabolites reached the plateau already 3 h after application of the patches and remained on extended and fairly constant levels during the whole 24-h application. The plasma levels of the three nitrates were proportional to the strengths of the patches. The plasma levels of 1,2-GDN were about 6 times higher than those of GTN. The plasma levels of 1,3-GDN were similar to those of GTN. Upon removal of the patches the concentrations of the three nitrates fell to negligible values within 3 h, an important property when an intermittent GTN therapy is needed in order to avoid tolerance to the drug. The patches were well tolerated at the application site. For their good tolerability, small size and transparency, the new GTN patches are very patient friendly, a quality that improves compliance with the therapeutic regimen. PMID- 11050695 TI - Influence of the onion as an essential ingredient of the Mediterranean diet on arterial blood pressure and blood fluidity. AB - Mediterranean diet has got a favourable effect on life expectancy. One of the crucial components of the diet are onions. In an open and a randomized, placebo controlled, double-blind, cross-over phase-I study a spontaneous pharmacological effect 5 h after administration of an onion-olive-oil maceration capsule formulation on arterial blood pressure could be demonstrated in apparently healthy subjects. In addition to a decrease in arterial blood pressure, a significant reduction in plasma viscosity and haematocrit were observed. These results are indicating a vasodilative effect of the onion-olive-oil-maceration product. The stickiness of the platelets was reduced. The effects were stronger in subjects with reduced blood fluidity compared to those subjects with normal rheological parameters. PMID- 11050696 TI - Comparative bioavailability of two formulations containing atenolol and chlortalidone associated in a 4:1 fixed combination. AB - Atenolol (CAS 29122-68-7) and chlortalidone (CAS 77-36-1) are marketed associated in a 4:1 strength ratio (100/25 and 50/12.5 mg) for the treatment of hypertension. According to EU guidelines, the bioequivalence of one dosage strength can also cover additional strengths when the pharmacokinetics of a given drug is linearly related with the dose. The kinetics of atenolol is linearly correlated with the dose and chlortalidone has linear kinetics with doses < or = 100 mg. Thus this trial carried out on the 100/25 mg strength also covers the 50/12.5 mg strength. The trial was carried out on 18 healthy volunteers (9 males and 9 females) according to a single dose, two-period, two-treatment, two sequence study design with washout. Timed atenolol plasma concentrations and chlortalidone blood concentrations were used to assess primary pharmacokinetic parameters Cmax, tmax and AUC extrapolated to infinity by a non-compartmental model. The bioavailability of the two formulations was compared through the 90% confidence intervals (C.I.) of Cmax and AUC in accordance with operating guidelines. C.I. of chlortalidone were fully comprised in the 0.80-1.25 range. In the case of atenolol, which displayed a higher data dispersion, C.I. were comprised in the enlarged 0.70-1.43 range. Time to peak, tmax, did not show any statistically significant difference between the test and reference product with respect to both analytes. Pharmacodynamic measurements of the decrease in systolic blood pressure led to fully overlapping results with test and reference. The authors conclude that the test formulation should be considered bioequivalent with the reference with chlortalidone and in the borderline of bioequivalence with atenolol. As no safety problems were involved and pharmacodynamics led to overlapping results as between test and reference, the bioequivalence conclusion could be extended also to atenolol. PMID- 11050697 TI - Effect of activated human protein C on disseminated intravascular coagulation induced by lipopolysaccharide in rats. AB - Protein C is the zymogen of an anticoagulant serine protease and is converted to its active form (activated protein C: APC) by thrombin in the presence of thrombomodulin. APC plays an important role in regulating coagulation and fibrinolysis by inactivating not only blood coagulation factors Va and VIIIa but also type-1 plasminogen activator inhibitor (PAI-1). The aim of the present study was to examine the effect of a human APC product (designated as CTC-111), compared with that of heparin, on the disseminated intravascular coagulation (DIC) induced by lipopolysaccharide (LPS) in rats. LPS (1 mg/kg/h) infusion was performed through a femoral vein for 4 h. One-fifth amount of the total dosage of CTC-111 or heparin was injected into the other femoral vein, followed by a 4-h infusion of the remainder. Both CTC-111 (10,000-100,000 U/kg) and heparin (400 800 IU/kg) inhibited the decrease in platelet count and fibrinogen level equally. The prolonged activated partial thromboplastin time and prothrombin time observed in DIC rats were further elongated in both CTC-111- and heparin-treated rats. But, this prolongation was less in CTC-111-treated rats than in the heparin treated ones. Heparin inhibited the increase in fibrin and fibrinogen degradation products more prominently than CTC-111. On the other hand, CTC-111 strongly inhibited the increase in PAI-1 activity but heparin did not. These results suggest that CTC-111 may enhance fibrinolysis through its direct inhibitory effect on PAI-1. The parameters for liver or renal damage, i.e., plasma glutamic oxaloacetic transaminase (GOT), glutamic-pyruvic transaminase (GPT), creatinine (Cre) and blood urea nitrogen (BUN), were significantly increased by LPS infusion. Both CTC-111 (100,000 U/kg) and heparin (800 IU/kg) decreased the increase in GOT and GPT levels significantly, whereas neither affected the increase in Cre or BUN. From these results, the activation of the blood coagulation system might partially contribute to the progression of liver damage caused by LPS, and might be less involved in the progression of renal damage in this model. In conclusion, CTC-111 showed both anticoagulant and profibrinolytic activity in the LPS-induced DIC model without excessive prolongation of coagulation time. From these results, CTC-111 is expected to be a useful remedy for DIC without the risk of bleeding. PMID- 11050698 TI - Determination of efficacy of linotroban by inducing a reduction of renal inulin/para-aminohippuric acid clearances with the thromboxane A2 mimetic U-46619 in the conscious female rat. AB - Linotroban (CAS 141443-73-4), a potent and selective thromboxane (TXA2) receptor antagonist, is known as a novel antithrombotic agent. It is suggested that pharmacological inhibition of TXA2 synthesis or action merits continued evaluation in the treatment of a variety of renal diseases. The aim of this study was the determination of efficacy of this new TXA2 receptor antagonist by assessing its effect on the reduction in inulin and para-aminohippuric acid (PAH) clearances induced by the TX/prostaglandin endoperoxide mimetic U-46619 in the conscious female rats. The following doses (3, 10, 30 mg/kg/24 h) of linotroban mixed with 720 micrograms TX-mimetic U-46619/kg/24 h were administered via osmotic pumps at a delivery rate of 10 microliters/h, implanted s.c. during 72 h. Rats of the U-46619 group were administered 720 micrograms U-46619/kg/24 h alone as described above, controls received 3.5% NaHCO3, respectively. Inulin/PAH clearances were determined at the end of the 4-h clearance period (68 h-72 h). Summarizing the data, glomerular filtration rate (GFR) and PAH clearances were reduced significantly by U-46619. When linotroban (3, 10 or 30 mg/kg/24 h) was added to U-46619 the GFR and PAH clearance were reversed to values that showed no significant differences to the controls. PMID- 11050699 TI - Pharmacodynamic study of low molecular weight dermatan sulphate (Desmin) after a single subcutaneous administration in patients with renal insufficiency. AB - The pharmacodynamic pattern of low molecular weight dermatan sulphate (CAS 24967 94-0, Desmin-LMWDS) was studied in patients presenting chronic renal insufficiency. Three groups of six patients were defined according to their creatinine clearance: group 1, more than 50 ml/min, group 2 between 10 and 50 ml/min and group 3 lower than 10 ml/min (haemodialized patients). Desmin-LMWDS concentrations were determined with the Heptest assay and the chromogenic specific heparin cofactor II dependent anti IIa assay. In patients of group 1 affected by moderate renal insufficiency, the pharmacodynamic profiles were roughly comparable to those obtained in normal subjects. In the two other groups, the profiles were markedly modified by the renal insufficiency. The maximal concentrations were doubled and the areas under the time-activity curve were 4 fold higher in haemodialyzed (group 3) and severe renal insufficient patients (group 2) than in patients of group 1. The clearance of the anti IIa activity were 13.98 +/- 6.25 l/h; 4.12 +/- 2.64 l/h and 2.94 +/- 1.53 l/h and the half lives were 2.79 +/- 2.60 h, 6.15 +/- 4.02 h and 11.51 +/- 6.54 h in groups 1 to 3, respectively (p < 0.05). The Desmin-LMWDS clearance was directly correlated to the creatinine clearance (r = 0.8244, n = 18, p < 0.001). Thus, as for low molecular weight heparin, renal function plays a major role in the elimination of low molecular weight dermatan sulphate. PMID- 11050700 TI - Comparison of the hematological effects of a sustained release chitosan formulation of pentoxifylline with a commercial formulation. AB - In the present study the hematological effects of a sustained release chitosan formulation of pentoxifylline (CAS 6493-05-6) were examined and compared with those of a commercial product. The study was carried out on 12 healthy volunteers. Both formulations were tolerated well clinically. The results demonstrated no antiaggregatory effect of the two different formulations of pentoxifylline in platelet rich plasma. Both drugs resulted in a decrease of plasma fibrinogen levels. A remarkable side effect of the new formulation was mild basophilia, without any clinical problems. PMID- 11050701 TI - Nigella sativa L. oil protects against induced hepatotoxicity and improves serum lipid profile in rats. AB - The effects of 4 weeks oral intake of Nigella sativa L. (NS) oil on some liver function tests and D-galactosamine- or carbon tetrachloride-induced hepatotoxicity were investigated in male albino rats. In another series of experiments, the effect of the oil on serum lipid profile was examined in male spontaneously hypertensive rats of stroke prone strain and Wistar Kyoto rats. The study showed that daily administration of the oil per se (800 mg/kg orally for 4 weeks) did not adversely effect the serum transaminases (ALT and AST), alkaline phosphatase, serum bilirubin or prothrombin activity in normal albino rats. When the oil was given for 4 weeks prior to induction of hepatotoxicity by D galactosamine or carbon tetrachloride, it was able to give complete protection against d-galactosamine and partial protection against carbon tetrachloride hepatotoxicity. NS oil showed a favourable effect on the serum lipid pattern where the administration of the oil (800 mg/kg orally for 4 weeks) caused a significant decrease in serum total cholesterol, low density lipoprotein, triglycerides and a significant elevation of serum high density lipoprotein level. PMID- 11050702 TI - Gastrointestinal tolerability of ibuprofen administered in two pharmaceutical formulations. AB - Ibuprofen (CAS 15687-27-1) is a nonsteroidal antiinflammatory drug endowed with analgesic, antiinflammatory and antipyretic activity. The main side effect of ibuprofen and nonsteroidal antiinflammatory agents is addressed to disturbances of the gastrointestinal tract, like gastric pyrosis, gastric and intestinal damage. A pharmaceutical formulation of ibuprofen in fast melting tablets consisting in gastroprotected microgranules to be ingested without concomitant water intake (Cibalginadue Fast, hereinafter referred to as test) was compared in this trial with a formulation of ibuprofen in tablets (reference) on 18 healthy volunteers in terms of gastrointestinal and general tolerability. Both the formulations are present on the market. The two formulations were administered according to a two-period, two-formulation, two-sequence, cross-over design in repeated dose regimen to steady state with wash-out. The dose was 400 mg (2 strengths) b.i.d. over 7 days. Before, during and after each study period general tolerability was carefully checked from blood/urine biochemical parameters, adverse events experienced and vital signs. The target parameters were the gastric permeability to sucrose and the intestinal permeability to lactulose and mannitol, which were administered orally and assayed in the urine excreted during a 6-h period. Urinary excretion > 0.15% of sucrose and > 0.04 of the lactulose to mannitol ratio are considered expression of increased gastric and intestinal permeability, respectively. Three volunteers treated with the reference showed an increased gastric permeability > 0.15%. Neither other gastric nor intestinal increased permeability was detected. Occult blood in faeces was negative in all the cases. The incidence of adverse effects experienced was higher with the reference (9 volunteers) than with the test (5 volunteers). In details gastric pyrosis was experienced by six volunteers treated with the reference and only by two volunteers treated with the test. The whole tolerability was better with the test formulation than with the reference, even if these differences did not reach any statistically significant degree. The better tolerability of the test was attributed to its gastroprotection. PMID- 11050703 TI - Synthesis of 1-aldofosfamide-perhydrothiazines. AB - Aldofosfamide-perhydrothiazine derivatives are a new class of prodrugs which spontaneously, with half-life times of 2 to > 12 h hydrolyse to the corresponding aldophosphamide in aquous solution. Synthesis of 1-aldofosfamide-perhydrothiazine (N,N'-(2-chloroethyl)-phosphorodiamide-2-(2'-[4'-carboxy-1',3'- perhydrothiazinyl])-ethylester) and a derivative, in which one 2-chlorethyl group of the alkylating function is substituted by a mesyl-ethyl-group (N-(2 Chloroethyl)-N'-(methanesulphonylethyl)- phosphorodiamide-2-(2'-[4'-carboxy-1',3' perhydro-thiazinyl] )-ethylester), is described. PMID- 11050704 TI - Synthesis of certain pyrimido[2,1-b]benzothiazole and benzothiazolo[2,3 b]quinazoline derivatives for in vitro antitumor and antiviral activities. AB - Some new pyrimido[2,1-b]benzothiazole and benzothiazolo[2,3-b]quinazoline derivatives have been synthesized and tested for their antitumor and antiviral activities. Among therm, compounds 5c and 8d exhibited a broad spectrum antitumor activity with full panel (MG-MID) median growth inhibition (GI50) of 11.0 and 11.9 mumol/l, respectively. On the other hand, compounds 5c and 5d showed potential activity against Herpes simplex type-1(HSV-1) with 61 and 50% reduction in the viral plaques, respectively. The detailed synthesis, spectroscopic and biological data are reported. PMID- 11050705 TI - Study on the decomposition products of thiadiazinthione and their anticancer properties. AB - Study of the anticancer properties of thirty-four 3,5-disubstituted-tetrahydro-2H 1,3,5-thiadiazin-2-thione derivatives has been carried out by using cytotoxicity assays against HeLa, HT-29 and Hep G2 cells. The decomposition products of thiadiazinthione 1 m have been studied and their anticancer properties evaluated. PMID- 11050706 TI - Kinetic model for 5-fluorouridine degradation. Catalytic effect of 5 fluorouracil. AB - The effects of 5-fluorouridine (5-FUR, CAS 316-46-1) degradation products, 5 fluorouracil (5-FU, CAS 51-21-8) and D-ribose (CAS 50-69-1), on its degradation rate was investigated following a 2(3) factorial design. The experimental data fitted to the proposed mathematical model which includes two parallel degradation mechanisms: the first one, a second order bimolecular reaction involving both 5 FUR and 5-FU, and the second, a first order one. Experimental data obtained show a high variability. Both graphic and statistical analysis of the experiments for which a full kinetic model was applied manifested that the degradation mechanism included an autocatalytic route and confirmed the role of the 5-FU on the hydrolysis of 5-FUR. PMID- 11050707 TI - One-month safety study of intraperitoneal VRCTC-310-Onco (crotoxin + cardiotoxin) in rats. AB - To evaluate the toxicity of VRCTC-310-Onco (Crotalus durissus terrificus crotoxin + cardiotoxin from Naja naja atra), 10 Sprague-Dawley rats were implanted with intraperitoneal slow-release devices and subjected to treatment with 0.5 microgram/g body weight/d for 14 days. Biochemical evidence at days 7 and 14 showed blood, muscular, renal and metabolic disturbance, mostly reversed by day 28. No significant changes were found in necropsy. The limited toxicity of i.p. VRCTC-310-Onco in rats deserves further study. PMID- 11050708 TI - Effect of Schistosoma mansoni infection and treatment on drug metabolizing enzymes. AB - The effect of Schistosoma mansoni infection on drug-metabolizing enzymes was investigated before and after treatment of S. mansoni-infected male albino mice with the antibilharzial drug praziquantel (CAS 55268-74-1). The drug was given in a total dose of 1000 mg/kg, and was administered at 500 mg/kg of body weight for two consecutive days each of 500 mg/kg of body weight. The drug was given 49 days after infection with 80 Egyptian strains of S. mansoni cercariae/mouse. The hepatic content of cytochrome P-450, cytochrome b-5 and NADPH cytochrome P-450 reductase were determined at 4, 8, 24, 72 h, one and two weeks after the second dose of treatment. The enzymes were determined in the microsomal fraction after homogenization and ultracentrifugation at 105,000 x g. The results indicate a marked decrease of most enzymes in the infected groups compared to normal controls. Two weeks after treatment there was an improvement of the level of most enzymes towards the normal values. The levels of liver function tests were also improved. Concerning the oogram pattern, there was a rapid change even after 8 h after the second dose of treatment. It was concluded that the antibilharzial drug could be considered as a safe drug with a rapid onset of action. PMID- 11050709 TI - Medical illustration. The impact of photography on its history. AB - In this article, developed from an oral presentation given at the thirty-seventh annual meeting of the B.P.A. in August, 1967 and first published in 1968 in the Journal of the Biological Photographic Association, some of the important stepping stones in the evolution of medical illustration are looked at--from the anatomists and artists of the fourteenth and fifteenth centuries, through the introduction of the camera obscura in the eighteenth century, and on to the advent of photography. PMID- 11050710 TI - Twenty-five years ago in JBPA/JBP. PMID- 11050711 TI - The use of digital photography to support a medical mission to Honduras. AB - The South Texas Physicians Outreach (STPO) organization makes at least one annual mission to Santa Rosa de Copan, Honduras to provide much needed medical and surgical care to the residents of western Honduras. The author was named the team photographer for both a trip in 1998 and a subsequent visit in 1999. In preparing for the 1999 trip, a plan to reduce the expenditure by STPO for photography support was put into motion. It was decided that digital photography would eliminate most of the cost for documentation of the team's activities in clinics, surgery, and human interest activities. This article provides a short history of the organization, a brief overview of the 1998 trip, and a discussion of the photographic aspects of the 1999 trip. PMID- 11050712 TI - Amino acid analysis. An overview. PMID- 11050713 TI - Amino acid analysis, using postcolumn ninhydrin detection, in a biotechnology laboratory. PMID- 11050714 TI - Purification of proteins using UltraMacro spin columns or ProSorb sample preparation cartridges for amino acid analysis. PMID- 11050715 TI - Amino acid analysis using precolumn derivatization with 6-aminoquinolyl-N hydroxysuccinimidyl carbamate. PMID- 11050716 TI - Amino acid analysis by high-performance liquid chromatography after derivatization with 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide (Marfey's reagent). PMID- 11050717 TI - The analysis of amino acids using precolumn derivatization, HPLC, and electrochemical detection. PMID- 11050718 TI - Anion exchange chromatography and integrated amperometric detection of amino acids. PMID- 11050719 TI - Ion-pair chromatography for identification of picomolar-order protein on a PVDF membrane. PMID- 11050720 TI - Capillary gas chromatographic analysis of protein and nonprotein amino acids in biological samples. PMID- 11050721 TI - Measurement of blood plasma amino acids in ultrafiltrates by high-performance liquid chromatography with automatic precolumn O-phthaldialdehyde derivatization. PMID- 11050722 TI - Determination of amino acids in foods by reversed-phase high-performance liquid chromatography with new precolumn derivatives, butylthiocarbamyl, and benzylthiocarbamyl derivatives compared to the phenylthiocarbamyl derivative and ion exchange chromatography. PMID- 11050723 TI - Amino acid measurement in body fluids using PITC derivatives. PMID- 11050724 TI - Determination of proteins, phosphatidylethanolamine, and phosphatidylserine in lipid-rich materials by analysis of phenylthiocarbamyl derivatives. PMID- 11050725 TI - Analysis of O-phosphoamino acids in biological samples by gas chromatography with flame photometric detection. PMID- 11050726 TI - Determination of sulfur amino acids, glutathione, and related aminothiols in biological samples by gas chromatography with flame photometric detection. PMID- 11050727 TI - Capillary electrophoretic determination of 4-hydroxyproline. PMID- 11050728 TI - Total plasma homocysteine analysis by HPLC with SBD-F precolumn derivatization. PMID- 11050729 TI - Determination of early glycation products by mass spectrometry and quantification of glycation mediated protein crosslinks by the incorporation of [14C]lysine into proteins. PMID- 11050730 TI - [Always prepared--good service or exaggerated servility in psychiatry?]. PMID- 11050731 TI - [Aggressive behavior of mentally incompetent psychiatrically ill criminals during inpatient treatment]. AB - OBJECTIVE, METHODS: To assess the amount of violent behaviour among mentally ill offenders NGRI during forensic long-term inpatient-treatment we retrospectively investigated the official incident-reports concerning verbal and physical aggression and damage to foreign property during an 8-year period in Austria's central high-security institution. RESULTS: 29.2% of our patients exhibited violent behaviour with only 7.8% being responsible for 41.1% of all incidents. Mentally impaired patients were significantly overrepresented in the violent group. Physical violence was reported in 25.8% (= 16.48 assaults/100 patients/year). 68% of the total amount of physical violence was directed against fellow patients. Violent behaviour was less driven by psychotic symptoms but rather by current everyday conflicts and stress situations. CONCLUSIONS: The majority of incidents had only minor consequences. Yet, an inquiry concerning the offenders' intentions and the danger experienced by staff members indicated a reasonable violent potential also in minor assaults which appears to be important with respect to ward climate and distress of staff. PMID- 11050732 TI - [Treatment motivation of patients admitted for substance abuse treatment according to the section 64 StGB regulation]. AB - OBJECTIVE: Describe treatment motivation in offenders with addiction problems, who enter treatment in forensic hospital departments according to section 64 of the German penal code. Consider relevance of treatment motivation with regard to course and outcome of treatment. METHOD: 83 patients were interviewed within the first days of psychiatric detention and after six month of treatment. They also filled in questionnaires on treatment motivation. Hospital staff reported another six month later about course of treatment. RESULTS: Most patients express awareness of drug/alcohol related problems and an intention to participate in treatment. But only a minority of patients shows a very strong motivation to stay abstinent from alcohol/drugs. More than half of the patients relapse during the first year of treatment. In more than one third of the cases, psychiatric detentions are cancelled due to a negative assessment of prognosis of further treatment. Most of these patients are returned to prison. CONCLUSIONS: Initial treatment motivation shows some relevance with regard to course of treatment. But negative outcomes of treatment shouldn't be explained simply as effects of insufficient motivation. A conception of treatment fostering hope and social competence seems to be most effective in strengthening treatment motivation. PMID- 11050733 TI - [Patient suicide during inpatient psychiatric therapy. New developments]. AB - The increase of suicides in the seventies and eighties during psychiatric hospital treatment is well documented. Suprisingly, results from the last years in the nineties show different courses of suicide rates in psychiatric hospitals, sometimes with a tendency of a decrease. PMID- 11050734 TI - [Are coercive measures carried out arbitrarily?]. AB - BACKGROUND: Due to media reports and psychiatric literature coercive measures (seclusion, restraint, forced medication) seem to be carried out rather arbitrarily in psychiatric hospitals. We examined whether associations between severity of psychopathological symptoms, patients' aggressive behaviour and coercive measures would be found. METHOD: 193 consecutive treatment episodes of our department for general psychiatry were examined. The psychopathological status at admission was recorded with the PANSS, aggressive behaviour during in patient treatment with the Modified Overt Aggression Scale (MOAS). For each treatment episode we recorded whether coercive measures were carried out or not. RESULTS: Coercive measures were carried out in 47 patients (24.3%). MOAS scores in these patients were six fold higher than in patients who were not exposed to coercive measures. 56% of coercive measures occurred in the quarter of patients with the highest psychopathology, none in the quarter with the lowest psychopathology (p < 0.0001). CONCLUSION: Contrary to frequent presumptions, the application of coercive measures in psychiatric hospitals is predominantly indicated by patients' psychopathology and violent behaviour. PMID- 11050735 TI - [What do medical students in the 6th clinical semester know about psychiatric emergencies?]. AB - OBJECTIVE: The aim of the present study was to evaluate the knowledge of medical students in psychiatric emergencies. METHODS: Medical students were questioned concerning their knowledge about psychiatric emergencies, shortly before finishing their theoretical part of the medical training program. RESULTS: Findings indicated that medical students have significant deficits in psychiatric knowledge. Nevertheless, most students reported to be interested in psychiatry and to acknowledge the importance of psychiatric knowledge for general practioners. CONCLUSIONS: The reasons for the reported deficits cannot be sufficiently explained by a lack of interest in psychiatry or a negation of the importance of knowledge in psychiatry for general practioners and may hint to conceptual problems in the organisation of the medical training programs. PMID- 11050736 TI - [Do cognitive deficits in depressive disorders remit?]. AB - OBJECTIVE: The importance of cognitive deficits concerning schizophrenics is well known. Much less material of that kind has been collected from patients with affective disorders, which was the aim of this clinical study. METHODS: Inclusion criterions were in- and outpatients with affective disorders (ICD-10: F31-F33) aged < 61. The clinical status was determined by use of the HAMD-21, the subjects were examined by means of standardised computerised cognitive performance tests (CGT-[M], DAUF). The results were correlated with the HAMD and the psychiatric pharmacotherapy. RESULTS: In each of the three subtests the heavily depressive group (HAMD > 24, n = 14) came off significantly (Mann-Whitney-U-Test, p < 0.05) worse than the remitted group (HAMD < or = 8, n = 18) while 8 of the 18 remitted patients showed pathological results in the CGT-(M). Only patients who took tranquilizers performed significantly worse than patients without such medication (p < 0.05). CONCLUSION: In some cases of affective disorders, a "cognitive residual syndrome" persists which is rather part of the disease than pharmacotherapy-associated. PMID- 11050737 TI - [Delirium syndrome as a side-effect of lithium in normal lithium levels]. AB - Lithium is used with great success in the treatment of manic patients and for prophylaxis of bipolar disorders. There are only few reports about neuropsychiatric side effects at therapeutic serum levels. We report on a 38 year old woman with bipolar disorder who was treated with lithium for 20 years without side-effects. Subsequent to a manic episode, she became disoriented at night and showed marked memory deficits. The patient did not show any neurological or gastrointestinal signs of intoxication. Lithium serum-levels were in therapeutic range. The psychiatric symptoms disappeared when lithium was stopped. We interpret these symptoms as delirant syndrome with pseudo-dementia at therapeutic lithium serum levels. This side-effect must be taken into account even in patients on successful longtime lithium therapy. PMID- 11050738 TI - [Subacute encephalopathy with epileptic seizures in an alcoholic patient]. AB - We introduce a case of a 66 year-old male with chronic alcoholism who suffered from confusion, Wernicke-aphasia and epileptic seizures. Several EEG revealed periodic lateralized epileptiform discharges. The patient's case resembles the symptoms of a subacute encephalopathy with epileptic seizures which can occur in alcoholics. PMID- 11050739 TI - [Psychiatric problems after implantation of a cardioverter/defibrillator (ICD)]. AB - The growing number of ICD patients diagnosed with malignant arrhythmias are often exposed to severe psychosocial stressors. In a number of cases, these stressors could lead to anxiety and depression. Although emotional stressors may be a major contributing factor in malignant arrhythmia, it is unfortunate that these symptoms are largely ignored. Therefore, additional psychiatric and psychotherapeutic treatment services for the prevention of sudden cardiac death seem advisable in certain cases. PMID- 11050740 TI - [Suicide in the family--heredity or imitation?]. PMID- 11050741 TI - [Proteases in helminthic parasites]. AB - Proteases catalyse the cleavage of internal peptide bonds within peptides and proteins. They are classified into four major classes and are involved in a broad range of eukaryotic processes. Proteases have also been found to play a number of critical roles in the virulence of pathogenic agents, particularly of nematode parasites. Parasitic proteases are involved in different aspects of host-parasite interactions. They facilitate the invasion of host tissues and allow nutrition as well as the survival of the parasite in its host. Proteases also participate in the parasite's evasion from the host's immune response. The functional diversity and complexity of these enzymes are described in this review, with a particular focus on the principally identified proteases of four helminths: Schistosoma sp., Fasciola sp., Taenia sp. and Haemonchus sp. Some of these proteases, especially the cysteine proteases secreted by the parasitic trematode Fasciola hepatica, have been successfully tested in experimental immunodiagnosis. Proteases identified in different parasites are currently under study for a use as recombinant vaccines. In this respect, proteases are proposed as major potential targets for immunotherapy and chemotherapy against parasitic diseases. PMID- 11050742 TI - Production of muraminidase-released protein (MRP), extracellular factor (EF) and suilysin by field isolates of Streptococcus suis capsular types 2, 1/2, 9, 7 and 3 isolated from swine in France. AB - A total of 323 isolates of Streptococcus suis recovered from diseased or healthy pigs in France were serotyped. The presence of virulence-related proteins, Muraminidase-Released Protein (MRP), Extracellular Factor (EF) and Suilysin was also studied in 122 isolates of capsular types 2, 1/2, 9, 7 and 3 to evaluate their implication in virulence of S. suis. Capsular types 2, 1/2, 9, 7 and 3 were the most frequently detected (93%), with 69% for the capsular type 2 alone. Capsular types 2, 1/2, 9, 7, 3, 1, 4, 8, 18, 10 and 12 were isolated from diseased pigs, whereas types 2, 7, 9, 1/2, and 3 originated from the nasal cavities or tonsils of healthy animals. Most of the S. suis type 2 isolates recovered from diseased pigs carried MRP+ EF- Suilysin- (46%) or MRP+ EF+ Suilysin+ (28%) phenotypes. The MRP+ EF- Suilysin- phenotype was also detected in 67% of S. suis type 2 strains isolated from healthy pigs. The production of the virulence-related proteins was less frequently found in S. suis types 1/2, 9, 7 and 3 recovered either from diseased or healthy pigs. In this study, all the capsular type 1/2 strains were MRP+ EF- Suilysin- and all the S. suis type 7 harboured an MRP- EF- Suilysin- phenotype. The MRP- EF- Suilysin- phenotype was found in S. suis types 2, 3, 7 and 9 isolated from septicaemia, meningitis, pneumonia, and pleurisy. These results suggest that the presence of these proteins should not be used as a single condition for classifying the virulence of a field isolate in France. PMID- 11050743 TI - Development of a complete ELISA using Salmonella lipopolysaccharides of various serogroups allowing to detect all infected pigs. AB - Although poultry is recognized as the major source of food-poisoning caused by Salmonella, pork also contributes to human infections. This study was therefore undertaken in order to develop a reliable serological method for the evaluation of the Salmonella status of piglets. A complete ELISA was performed using lipopolysaccharides of Salmonella Typhimurium, Anatum, Hadar and Infantis because these serovars were representative of the serogroups isolated from 30 contaminated fattening farms. S. Enteritidis was also added because of its importance in human infection and to include the O:9 antigen. This method potentially detects 100% of infected pigs. A significant correlation was found between this serological method and the bacteriological data from mesenteric lymph nodes (p = 0.01). In addition, both sensitivity and specificity were high (97% and 94% respectively). The ELISA test was therefore used in a cross sectional study on 4 farms to evaluate when pigs became contaminated: seropositive pigs were only found for the 20 week old finishing pigs. The antibody response to Salmonella in piglets was also investigated: maternal antibodies persisted until 7 weeks of age and post-Salmonella contamination seroconversion was detected from 8 weeks of age onwards. PMID- 11050744 TI - Enzyme-linked immunosorbent assay with a Salmonella enteritidis antigen for differentiating infected from vaccinated poultry. AB - The specificity and sensitivity of indirect ELISA, based on the use of four different antigenic extracts obtained from a clinical isolate of Salmonella enteritidis, were compared with those obtained with the gm-flagellin based ELISA (IDEXX). A total of 116 serum samples from salmonellae free, naturally infected and vaccinated hens were studied. The results showed that the indirect ELISA, based on lipopolysaccharide (LPS), O-polysaccharide (PS) or membrane sediment (SD) antigens, enable the identification of a greater number of infected birds and discriminated field antibody responses from vaccinal ones better than the commercial IDEXX test. The indirect ELISA that used a O-polysaccharide rich fraction (PS) proved to be the most specific and sensitive test, suggesting that this indirect ELISA could be used to confirm IDEXX results, especially when the differentiation between vaccinated and infected poultry is required. PMID- 11050745 TI - The urinary excretion of selenium in sheep treated with a vasopressin analogue. AB - The renal excretion of selenium was investigated in ewes with an excretion of hypotonic urine (control group) and in ewes with a formation of highly concentrated urine. Chronic stimulation of the urinary concentrating activity of sheep kidneys was induced by a long-term treatment with 1-desamino-8-D-arginine vasopressin (dDAVP), a synthetic analogue of vasopressin with prolonged effects. Young animals with 22 to 25 kg b.w. were fed a normal protein diet providing a daily intake of 129.25 g of crude protein, 12.03 MJ of digestible energy and 0.18 mg of selenium for 3 weeks. The vasopressin treated sheep (n = 11) were given subcutaneous injections of 12.5 micrograms of dDAVP in glycerol twice daily for one week before the clearance measurement of renal functions. The control group (n = 11) was treated with glycerol only. The administration of dDAVP resulted in a highly significant decrease of the urinary flow rate (from 3.19 +/- 0.50 in control group to 0.33 +/- 0.03 mL.min-1 in dDAVP animals, P < 0.001) without changes in the glomerular filtration rate (80.18 +/- 6.36 in controls vs. 77.86 +/- 6.26 mL.min-1, NS). No effects on plasma selenium level were observed (0.17 +/- 0.03 in controls vs. 0.20 +/- 0.03 mumol.L-1, NS) but the amounts of selenium excreted were found to be highly significantly reduced (from 0.29 +/- 0.05 in controls to 0.03 +/- 0.01 nmol.min-1, P < 0.001) in dDAVP treated sheep. Despite a large reduction in urinary flow rate, the selenium concentration in urine was actually the same in both groups (0.09 +/- 0.01 mumol.L-1) resulting in a sharp fall in the renal clearance of selenium (2.20 +/- 0.54 in controls vs. 0.18 +/- 0.03 mL.min-1, P < 0.01) due to dDAVP. This seems to be a consequence of the large increase in the selenium solvent drag induced by a vasopressin treatment. The results presented suggest that vasopressin may contribute to maintenance of the selenium balance in sheep via its effects on renal function. PMID- 11050746 TI - Metalloproteinases and tumor necrosis factor-alpha activities in synovial fluids of horses: correlation with articular cartilage alterations. AB - Early detection of osteoarthritis in horses represents a challenge for equine practitioners. Several biological markers have been implicated in the pathological processes involved in articular cartilage destruction. To further document cartilage matrix proteases production, synovial fluid was collected from 14 horses (90 joints) before they were subjected to euthanasia. Growth macroscopic examination of the joints gave information on cartilage alterations. Samples were analyzed for matrix metalloproteinase (MMPs) activities by gelatin zymography and tumor necrosis factor alpha (TNF-alpha) cytotoxicity using L929 cells. Significant increase of MMP-9 monomer and dimer were found in synovial fluids of joints with severe cartilage alterations. On the contrary, the activity of TNF-alpha was not correlated to the degree of joint damage. The levels of MMP 9 monomer and dimer in the synovial fluid could reflect cartilage alteration in arthritis in the horse. PMID- 11050747 TI - Pharmacokinetics of sulphadoxine and trimethoprim and tissue irritation caused by two sulphadoxine-trimethoprim containing products after subcutaneous administration in pre-ruminant calves. AB - The pharmacokinetics of sulphadoxine-trimethoprim was studied in 6 pre-ruminant calves using two different products. Product A, which contained 200 mg sulphadoxine and 40 mg trimethoprim per mL, was administered intravenously or subcutaneously at a dosage of 25 mg sulphadoxine and 5 mg trimethoprim.kg-1 bodyweight. Product B, containing 62.5 mg sulphadoxine and 12.5 mg trimethoprim per mL plus lidocaine (1 mg.mL-1), was given subcutaneously at the same dosage. After intravenous administration of product A the mean time of half-life of elimination phase (t1/2) for sulphadoxine was 12.9 h, steady-state volume of distribution (Vd(ss)) was 0.44 L.kg-1 and clearance was 0.024 L.kg-1.h-1. Respective values for trimethoprim were 1.9 h, 2.0 L.kg-1 and 0.9 L.kg-1.h-1. After subcutaneous administration, the bioavailability of sulphadoxine was 96% and 98% and the time to reach a maximum concentration was 6.3 and 8.0 h for products A and B, respectively. The Cmax for trimethoprim was higher for product A (0.49 microgram.mL-1) than for product B (0.32 microgram.mL-1) (p = 0.014). Slow absorption from the injection site appeared to delay the elimination of trimethoprim after subcutaneous administration when compared to that after intravenous administration: apparent elimination t1/2 for trimethoprim after intravenous administration of product A was 1.9 h compared to 3.9 h and 3.6 h after subcutaneous administration of products A and B, respectively. The difference between intravenous and subcutaneous administrations was statistically significant (p < 0.05). Also the mean residence time was significantly shorter (p < 0.05) after intravenous administration (2.4 h) than that after subcutaneous administration of product A (6.9 h) and B (7.1 h). The bioavailability of trimethoprim was lower than that of sulphadoxine: 76% and 74% for products A and B, respectively. All 6 calves showed pain after subcutaneous administration of product A and the injection sites were warm and showed soft oedematous reactions 5-8 cm in diameter. Three of the calves also showed some pain after subcutaneous administration of product B; the local reactions were less severe. A marked increase was seen in creatine kinase activity after subcutaneous administration of both products. Product A caused a more pronounced increase but the difference was not statistically significant. We suggest 30 mg.kg-1 at 24-h intervals or alternatively 15 mg.kg-1 at 12-h intervals as the minimum dosage of sulphadoxine trimethoprim combination for pre-ruminant calves. Extravascular routes of administration should be avoided due to marked tissue irritation at the injection site. PMID- 11050749 TI - [Pelvic injury]. PMID- 11050748 TI - Immunisation of lambs with excretory secretory products of Oestrus ovis third instar larvae and subsequent experimental challenge. AB - Excretory-secretory products (ESP) of myiasis producing agents are involved in nutrition and development of larvae and are often immunogens. This study was carried out in order to define the antigenicity, the immunogenicity of Oestrus ovis ESP and the role of sheep immune response to ESP. Twenty-four six to eight month old female lambs were randomly allocated into two groups. The first one was immunised twice, four weeks apart, with excretory-secretory products of Oestrus ovis third instar larvae (L3ESP) in complete then incomplete Freund adjuvant. The second one served as a control, and received two injections of PBS plus complete and incomplete Freund adjuvant. Fifteen and twenty-eight days after the second immunisation, animals of both groups were experimentally challenged with O. ovis first instar larvae. Twelve days after the second experimental challenge, the twenty-four lambs were necropsied. The total number of O. ovis larvae, their stages of development, weights and sizes were recorded per animal and compared between the two groups. Establishment rates were very similar in both groups: 39% and 35% in control and vaccinated groups respectively but the percentage of developing stages was higher in the control group (13%) than in the vaccinated group (6%). It was concluded that the L3ESP immunisation of sheep did not protect against larval establishment but provided an inhibitory effect on larval growth. PMID- 11050750 TI - [Injuries and being injured. Hidden wounds after accident injuries]. AB - Motor vehicle accidents may cause apart from physical also psychological aftermath. Despite of posttraumatic psychologic symptoms may remit after 2-3 months, 10-20% of the injured persons will develop sustainable psychopathological symptoms. This report aims at an overview on possible psychopathological disturbances and their perpetuating conditions. Possible treatments are shown and the necessity of an interdisciplinary cooperation with the aim of a secondary prevention is emphasized. PMID- 11050751 TI - [Classification, staging, urgency and indications in pelvic injuries]. AB - For primary evaluation, classification and indication of pelvic ring injuries the exact knowledge of the injury mechanism and the clinical and radiological signs is mandatory. Clear injury definitions are proved for prognostical reasons and for the timing of the specific treatment. The simple classification of stable A type injuries, rotational B-type injuries and translationally stable C-type injuries is the basis for further treatment. Whereas A-type fractures normally need no surgical stabilization, except in severely displaced fractures or possible organ injuries due to fracture fragments, in B-type injuries solely stabilization of the anterior pelvic ring provides sufficient stability for early ambulation with partial weight bearing. In C-type injuries a combined posterior and anterior stabilization is required for anatomical reduction and early ambulation. With this concept the pelvic girdle can be reconstructed anatomically in the majority of cases. PMID- 11050752 TI - [Acetabulum fractures. Diagnosis, classification, evaluation]. AB - Diagnostics of acetabular lesions rest on careful explored medical history, clinical findings and X-ray-examination. Apart from standard radiography (pelvis a. p., injured hip a. p., iliac-oblique and obturator-oblique), which enables classification and supports strategic therapy decisions, computed tomography is considered indispensable by the majority. Fragment interposition, marginal impaction and concomitant occult pelvic ring fractures are detected and are the basis for special tactical decisions in operative treatment. The classification of Letournel with 5 elementary and 5 associated fracture types is still widely used. The AO-classification gives a solid basis for scientific evaluation and is a fundamental for a therapeutic concept with its basic structure with types and groups, ascending according to damage severity. Evaluation of acetabular fractures has to include additional and concomitant lesions and expected late damages. Only an anatomic reconstruction of the weight bearing part of the socket gives a chance for functional restitution. Osteoarthritis and disabling ectopic ossifications are not always avoidable and are subsequently the cause of impairment and respectively loss of working ability. PMID- 11050753 TI - [Injuries of the pelvic girdle. The pathway to exact diagnosis: which imaging methods are indicated? Synopsis of information]. AB - Synopsis in diagnosis of pelvic ring fractures begins with the help of our senses while taking the history, inspecting and palpating the patient. It is continued with urological, gynecological and neurological assessment of the whole extent of peripelvineal injury. The picture is completed by ultrasonography, standard X rays and special views, two- and three-dimensional CT and finally MRI. To minimize radiation damage to the patient and to lower the costs for radiologic assessment the standard ap view of the pelvis has to be analysed carefully to avoid numerous special views. Horizontal lines drawn perpendicular to the L5 Sacrum-Symphysis axis are helpful in discriminating rotational and translational instability, quantifying the extent of dislocation and controlling the quality of reduction and internal fixation. In order to avoid missing fractures of the os sacrum, the arcuate lines have to be inspected carefully. PMID- 11050754 TI - [Expert assessment of pelvic injuries]. AB - Progress in diagnostics and therapy of injuries of the pelvis made in the last few years also brought advantages in the assessment of longterm outcome. Despite correct anatomical reconstruction of the pelvic-ring lumbal and sacral pain is still a problem. The intensity of pain doesn't correlate with the grade of injury. So effects of pelvic fractures on static and dynamic of the musculoskeletal system and especially the function of the hip-joint can be assessed clearly. Furthermore neurological examinations done by specialist to find out specific deficits are of utmost importance. Advanced radiological and technical diagnostics like computer tomography are significant for the assessment of pelvic injuries as well as the clinical examination. PMID- 11050755 TI - [Early surgery after hip para-articular femoral fracture. Results of a prospective study of surgical timing in 161 elderly patients]. AB - In a prospective analysis the question should be answered, wether the mortality rate of femur fractures close to the hip joint can be diminished by operating as early as possible. 161 patients elder than 65 years could be included in the study. 86% of the 161 patients were operated upon 24 hours after trauma. The infection rate amounted to 3.4% after endoprothesis and to 1.2% after osteosynthesis. The hospital mortality was 7.4%. More than 85% of the patients could be discharged into the usual domestic surroundings. The rate of systemic complications (25.3%) was similar to the rate reported by the Chamber of Physicians with 26.1%. The hospital length of stay could not be diminished by this concept. By operating as early as possible the patients' request for mobility is fulfilled without running unjustifiable risks regarding mortality and postoperative complications. The mortality rate corresponds to the literature. PMID- 11050756 TI - [Treatment outcome after primary hemi-alloarthroplasty in dislocated humeral head fractures]. AB - Humeral head fractures often lead to a high complication rate because of vulnerable vascularization of the humeral head, pretraumatical soft tissue lesions and osteoporotic bone. Compared with open reduction and internal fixation primary hemiarthroplasty seems to be an encouraging alternative treatment in case of dislocated and comminuted fractures. To evaluate our results following primary hemiarthroplasty we examined the outcome of 27 patients treated between November 1993 and May 1997 by primary hemiarthroplasty (15 Neer II, 12 Aequalis) in our institution within two weeks after trauma. The average patient follow-up was 3.5 years. Postoperatively more than 80% of the patients reported no pain. Excellent or good results with abduction over 90 degrees, low or moderate loss of power and low restraint in activities of all day living were achieved in about 50% of patients. The results according to the criteria of the Constant-Score depended on the intensity of the rehabilitation programme with an average outcome of 65 points. Maybe in the future functional results can be improved by a more aggressive rehabilitation and stronger rotator cuff refixation. PMID- 11050757 TI - [Unstable pediatric femoral and forearm shaft fractures. Comparison between conservative treatment and stable intramedullary nailing]. AB - Elastic intramedullary nailing represents a new surgical concept in the treatment of unstable shaft fractures in children. The present case control study wanted to examine the superiority of intramedullary nailing in comparison to conservative therapeutic concepts which had been applied so far. 13 children with forearm fractures who were treated initially by conservative measures were compared to 13 other children who received a primary intramedullary nailing. With femoral fractures, 12 children were included in each group. In each patient pair age, type and localisation of the fracture were comparable. During the observation period (until the termination of final therapeutic measures or until the third year after injury) we examined clinical variables and subjective findings. Both therapeutic concepts led to comparably good functional results. Also subjective judgement of the therapeutic success did not differ between groups. However, with intramedullary nailing of shaft fractures of the femur the mean hospital length of stay (7.0 +/- 3.5 days) was significantly shorter than with initial conservative treatment (36.5 +/- 2.2 days, P < 0.05). Irrespective of the localisation of the fracture intramedullary nailing required significantly less x ray examinations during the observation period. These results suggest intramedullary nailing to be the procedure of choice to treat unstable forearm and femoral fractures in children. PMID- 11050758 TI - [Pretherapeutic diagnosis of fibrous dysplasia]. AB - A safe differentiation of fibrodysplastic lesions from "real" bone tumours is of high importance because a fibrous dysplasia often requires no further therapy. While polyostotic involvement of fibrous dysplasia can be safely diagnosed before therapy, in monostotic disease differential diagnostic problems may occur. In the present investigation only in 6 of 14 mon- and biostotic lesions caused by fibrous dysplasia a correct diagnosis could be established by radiologic methods. However, in all cases of fibrous dysplasia malignancy could be excluded by radiology and the false diagnosis had no therapeutic consequences. PMID- 11050759 TI - [A great teacher and "upright neighbor". Eduard Albert on the 100th anniversary of his death]. AB - The surgeon Eduard Albert was unjustified standing in the shadow of his great contemporary neighbour in Vienna, Theodor Billroth, for a long time. Not before their career finished they payed each other respects. Autobiographical reminiscences of important disciples and historical works later generations made a contribution to the recognition of Albert's services and personality. PMID- 11050760 TI - [Preissler endoscopic carpal ligament release. Experiences from 1,000 operations]. AB - To reduce the postoperative morbidity different methods of endoscopic carpal tunnel release have been developed since 1989. We report our results and experiences with the method described by Preissler. Since its introduction in our clinic 1995, 1,000 patients have been operated until now. Out of 477 with an electrophysiological examination, 396 patients (83%) showed no symptoms. In three cases it was necessary to change to an open approach during operation, there were no other intraoperative complications. To evaluate our results we sent a questionnaire to 32 of our patients that were operated on one hand with an open technique and on the other hand with the endoscopic technique of Preissler. The results demonstrated that the endoscopic method was significantly better. From our experience endoscopic technique for carpal tunnel release Preissler's is a safe, well accepted, easy for learning and cheap alternative to the open procedure. PMID- 11050761 TI - [Arthroscopic therapy of Baker's cyst]. AB - Arthroscopic therapy of Baker's cysts contains repair of all intraarticular lesions and sealing of the junction between Baker's cyst and the dorsal recessus of the knee joint. All therapeutic steps can be sufficiently done throughout the ventral standard approaches. 18 patients have been treated by this technique between 1992 and 1994. 14 of them were examined after an average of 27 months. In all patients one or more lesions were detected arthroscopically. (12 cartilage damages, 10 synovialities, 9 meniscal tears, 2 ruptures of the a.c.l.). Cyst recurrence was found arthrosonographically in three cases. These results are comparable to that reported after open cyst extirpation. In our patients we did not see any intra- or postoperative complications. Therefore we consider the arthroscopic therapy of Baker's cyst to be a more comfortable alternative to open extirpation. PMID- 11050762 TI - [Induced abortions in the Third Reich. Legal basis and provision]. AB - This article analyses, after introductory comments on the legal situation in the German Empire and the Weimar Republic, the legal basis for induced abortions during National Socialist rule in Germany. During this period the first legal definition for eugenically and medically indicated abortions was established. At the same time the prohibition of induced abortions outside these criteria was controlled more strictly and violations were punished more severely. This concerned abortions mainly for social reasons. The intention was to legalize abortion for those deemed "less worthy" while, at the same time, to minimise the number of abortions of those considered as "more valuable" to society. The main thrust of this policy was to increase the birth rate of "valuable" citizens. The second part of this paper focuses on eugenic and medical abortions at the University of Freiburg's Maternity Hospital. PMID- 11050763 TI - [Hand-held ultrasound-guided vacuum biopsy of mammary lesions--first experiences]. AB - OBJECTIVE: Evaluation of indications for ultrasound guided hand-held Mammotome biopsy. MATERIAL AND METHODS: To achieve breast diagnosis 50 ultrasound guided hand-held Mammotome-biopsies were performed between January 3rd and April 4th, 2000. RESULTS: 34 patients presented with non-palpable, 16 with palpable breast lesions. The benign-to-malignant ratio was 80% to 20%. Complete removal of the lesion we established in 22 (44%) of all 50 procedures. Definitive breast diagnosis of malignancies was achieved in all but one case. CONCLUSIONS: We regard the ultrasound guided hand-held Mammotome-biopsy as diagnostic and surgical instrument that provides the clinician with a flexible and easy to use method of accurate breast diagnosis. Lesions too small, superficial, or deep for conventional core biopsy are indications for a ultrasound guided Mammotome-biopsy as well as abnormalities where wide sampling is considered important or small fibroadenomas. However, standard of care for breast diagnosis remains the conventional hand-held 14-gauge-core-biopsy. For malignant lesions hand held mammotomy must be regarded as a diagnostic and not a therapeutic procedure. PMID- 11050764 TI - [Skin sparing mastectomy with autologous immediate reconstruction: oncological risks and aesthetic results]. AB - OBJECTIVE: Is the oncological safety of skin sparing mastectomy (SSM) with immediate autologous reconstruction and improved aesthetic results comparable to postoperative findings in patients treated with modified radical mastectomy (MRM)? MATERIAL AND METHODS: Sixty patients with T1-2 breast carcinomas and contraindications for breast conserving therapy were treated by SSM and compared to 81 patients of the same age groups and MRM with regard to oncological and aesthetic data. In 33 (55%) patients the nipple areola complex (NAC) could be spared. For autologous tissue TRAM- and Latissimus dorsi-flaps were used. The mean follow-up was 40 (range 20-71) months. RESULTS: The observed local recurrence rates were not significantly different (p = 0.443) after SSM (n = 3; 5.0%) or MRM (n = 5; 6.2%). Distant metastases and death were seen in 26.7% and 15.0% (SSM), respectively, and in 25.9% and 13.5% (MRM), respectively. Body mass index, operation time and postoperative haemoglobin concentration differed between both groups significantly (p < 0.001) but not the rate of complications (p = 0.232). Aesthetic results of SSM were judged as excellent or good in 90.0% of patients and in 83.4% of surgeons. Nine patients (11.1%) underwent a secondary breast reconstruction after MRM. Furthermore, 12 (14.8%) patients with MRM would prefer a SSM with immediate reconstruction in a similar situation. CONCLUSION: Skin-sparing mastectomy improves aesthetic results to a high degree without increasing of local or distant recurrence rates. Skin-sparing mastectomy should be offered to selected patients with breast cancer as an alternative to modified radical mastectomy. PMID- 11050765 TI - [Parameters of lipid metabolism in postmenopausal women on oral or transdermal hormone replacement with or without progesterone]. AB - OBJECTIVE: Aim of the study was to show different influences of transdermal and oral hormone replacement therapies (conjugated and micronized estrogens) with or without varying dosages of C21-progestogens on serum lipids and lipoproteins. MATERIAL AND METHODS: We report on serum triglycerides, cholesterol, HDL-, LDL cholesterol, apolipoprotein A 1 and B levels of 80 postmenopausal women, who received hormone replacement therapies for more than one year. RESULTS: All patients showed increasing (non-significant) serum HDL/LDL-cholesterol-ratios. Transdermal estrogen monotherapy also influenced the lipid parameters in a positive way. Apolipoprotein B, cholesterol and LDL-Cholesterol decreased, apolipoprotein A 1 increased. Transdermal replacement therapy combined with C21 progestagens and all oral therapies resulted in HDL-cholesterol increases. Positive changes in lipid parameters were most remarkably in women receiving oral therapies. The addition of 42 mg medrogestone/cycle caused a more significant decrease of cholesterol serum levels than higher dosages of medrogestone did. During subsequent treatment cycles, serum triglycerides showed increasing levels within the reference limits in women receiving conjugated estrogens and medrogeston. CONCLUSIONS: Transdermal and oral hormone replacement therapies with and without C21-progestogens are ideal for hormone replacement therapy in postmenopausal women with normal or minor pathological lipid parameters. The lowest possible medrogestone dose necessary for endometrium protection should be used. PMID- 11050766 TI - [Prognostic value of Fallopian tube endoscopy]. AB - OBJECTIVE: Progress in sterility-therapy demands diagnostic methods, that ensure the right indication for certain therapies in order to avoid unsuccessful attempts. In this retrospective study we tried to find out, whether we could predict and influence the success rate of therapies with the use of falloposcopy prior to further treatment. MATERIAL AND METHODS: In 62 sterile patients the endotubal state was evaluated with a falloposcope (Imagyn, Irvine, USA), a system of linear everting catheter (LEC) and microendoscope, prior to further treatment. RESULTS: Twenty of the 62 patients had endotubal pathology of both tubes, 15 patients showed pathology of one tube and 25 patients were diagnosed to have a normal endotubal morphology. In 2 patients falloposcopy could not be performed because of blockage due to intramural myomas. Pregnancy rate of those patients with a normal endosalpinx was 52% altogether, following microsurgery 80% and following insemination even 100%. No extrauterine pregnancies were described in this group. Pregnancy rate of those patients with endotubal pathology of both tubes was 35%. In this group no pregnancy followed microsurgery and following insemination one extrauterine pregnancy could be noticed. Furthermore, IVF results were improved if morphologic evaluation showed normal mucosa. CONCLUSIONS: The falloposcopy is a helpful diagnostic procedure to evaluate endotubal pathology prior to microsurgery and insemination. PMID- 11050767 TI - Effect of corticosteroids on sperm antibody concentration in different biological fluids and on pregnancy outcome in immunologic infertility. AB - OBJECTIVE: In the last fourteen years, detection of free spermagglutinating antibodies has been performed by tray agglutination test (TAT) and by direct/indirect mixed antiglobulin reaction test (MAR) in 696 infertile couples aged 23-42 years (female) and 26-52 years (male) with previously undiagnosed infertility. MATERIAL AND METHODS: Oral decreasing doses of prednisone or dexamethasone for three months in the case of serum or seminal plasma spermantibodies, plasmapheresis in one patient with serum spermantibodies were used. Hydrocortisone to the ectocervix was applicated in patients with spermagglutinating antibodies in cervical ovulatory mucus. RESULTS: Serum IgG spermagglutinating antibodies totally disappeared in 3 out of 11 men, and four out of 15 women. But serum IgM sperm antibodies persisted. One female patient was treated with corticosteroids and also with plasmapheresis. Seminal plasma IgG spermagglutinating antibodies were greatly influenced in 57.8% of the patients, IgA in 38.9% and each case by oral corticosteroids. A decrease of IgG and IgA spermagglutinating antibodies in ovulatory cervical mucus during hydrocortisone local application was registered in 61.3%, IgG in 50.7% and IgA in 65.9%. Levels of IgM spermagglutinating antibodies in cervical ovulatory mucus were not influenced, spermantibodies in IgA and IgE were affected very little. CONCLUSION: The corticosteroid influence of immunocompetent cells in each case must be chosen individually with regard to the localisation of spermantibodies. We often combine the long lasting corticosteroids treatment with in vitro fertilization. The therapy seems to be very promising for the improvement of immunological causes of infertility. PMID- 11050768 TI - Clinical and community strains of Klebsiella pneumoniae: multiple and increasing rates of antibiotic resistance in Abha, Saudi Arabia. AB - Klebsiella pneumoniae is the most commonly isolated bacterial species in a maternity hospital in Saudi Arabia. Here, 380 strains isolated in 1997 and 480 strains in 1999 were studied for their resistance antibiograms, using the standardised disc diffusion test. Of 16 antibiotics tested, four in 1997 and six in 1999 were ineffective against > 50% of the respective isolates, and resistance rates to 11 antibiotics increased over the two-year period (P = 0.05-< 0.0001). With resistance rates of < 20%, imipenem, ciprofloxacin, gentamicin and cefotaxime were more effective in 1997; only imipenem and ciprofloxacin remained as effective in 1999. In addition, 105 community strains were tested and > 50% were resistant to four antibiotics. Resistance rates to most antibiotics were lower than those of clinical strains (P = 0.0285-< 0.0001). Imipenem resistance was detected among both clinical and community isolates. Multiresistance was 64.5% in 1997 and 79.2% in 1999 (P < 0.0001), and 83.8% in community strains in 1999. Using the double-disc synergy test, extended-spectrum beta-lactamase (ESBL) was detected in 27.5% of ceftazidime-resistant clinical strains isolated in 1999. Among the clinical strains, seven (65%) and 11 (67.9%) resistance antibiograms occurred frequently in 1997 and 1999, respectively. Such frequency was not observed among community isolates. These findings confirm the alarmingly high rates of multiresistance and the emergence of ESBL-producing strains, highlighting the urgent need to restrict over-the-counter availability of antibiotics, and increase awareness in the local medical community. PMID- 11050769 TI - Re-expression of procollagen type IIA in oral squamous cell carcinoma. AB - There are two forms of procollagen type II (IIA and IIB), both of which are expressed during chondrogenesis. Procollagen type IIA also is present at sites of developmental epithelial-mesenchymal interaction. Malignant transformation is associated with disturbed epithelial-mesenchymal interaction and the reappearance of fetal characteristics. This study aims to determine whether or not procollagen type IIA is re-expressed in oral squamous cell carcinoma (OSCC). Immunoperoxidase techniques were applied to frozen and paraffin sections of OSCC (n = 30) and normal oral mucosa (n = 5). In the carcinoma group, strong cytoplasmic staining for collagen type II was present (25/30). Staining was weak or absent in the stroma, and absent from the normal oral mucosa. Frozen sections from 10 of the carcinoma cases which showed positive staining were incubated with antibodies specific for procollagen type IIA and visualised using immunofluorescence. Staining was evident in each case and was particularly strong in the region of the basement membrane. Slot-blot analysis of collagen extracts from OSCC supported the immunohistochemical findings. We conclude, therefore, that procollagen type IIA is re-expressed in OSCC. PMID- 11050770 TI - Ergonomic intervention: its effect on working posture and musculoskeletal symptoms in female biomedical scientists. AB - This study investigates the effect of ergonomic intervention on working posture and musculoskeletal symptoms in female biomedical scientists. The Nordic musculoskeletal questionnaire (NMQ), body discomfort chart (BDC) and rapid upper limb assessment (RULA) are the tools for assessment. The study was conducted in three phases: pre-intervention, intervention and post-intervention. Pre intervention, 79% of subjects reported a three-month prevalence of symptoms, and these were reported more frequently by those working in haematology/transfusion. Analysis by RULA showed that the majority (59%) of postures had a grand score of four. A further 24% had scores of five or six. The highest frequency of poor postures was seen in haematology/transfusion. Intervention comprised physical workplace changes, a seminar, and advice on risk factors. In the post intervention phase, baseline measurements were repeated. Reporting of three-month prevalence of symptoms had decreased to 54%, and reports of body discomfort also had decreased. The majority (64%) had a RULA grand score of three. No observed postures had scores of five or six. In conclusion, ergonomic intervention resulted in an improvement in working postures, and a decrease in the prevalence of musculoskeletal symptoms and body discomfort. Analysis of findings indicate that RULA scores generally corresponded with reporting of symptoms (NMQ) and discomfort (BDC). PMID- 11050771 TI - Vascular endothelial growth factor and its receptor, Flt-1, in smokers and non smokers. AB - Raised levels of plasma vascular endothelial growth factor (VEGF) are found in some cancers, diabetes, and certain other conditions, but levels of its receptor, soluble Flt-1 (sFlt-1), in these diseases have yet to be reported. We hypothesised that smoking would influence levels of these molecules. Consequently, we measured VEGF and sFlt-1 by enzyme-linked immunosorbent assay (ELISA) in plasma from 92 non-smokers and 35 smokers. No difference in VEGF was seen between the groups but, despite considerable overlap, sFlt-1 was significantly lower in smokers (P = 0.027). VEGF and sFlt-1 correlated strongly with each other (P < 0.001). Although VEGF may arise from a number of cell types, including endothelial cells, the primary source of sFlt-1 is thought to be the endothelium; however, neither VEGF nor sFlt-1 correlated with levels of the endothelial cell activation/damage marker soluble thrombomodulin. Our data point to changes in levels of the VEGF receptor, sFlt-1--but not VEGF itself--in smokers, which appears to be unrelated to endothelial cell function. PMID- 11050772 TI - Long-term preservation of thermophilic Campylobacter species: a simple method. PMID- 11050773 TI - Vascular endothelial growth factor in human milk. PMID- 11050774 TI - Coeliac disease in adolescents/young adults: difficulties in monitoring. PMID- 11050775 TI - Detection of genital and cutaneous human papillomavirus types: differences in the sensitivity of generic PCRs, and consequences for clinical virological diagnosis. PMID- 11050776 TI - Extended-spectrum beta-lactamase-producing Klebsiella spp. AB - Extended-spectrum beta-lactamases (ESBLs) are associated particularly with Klebsiella spp. These enzymes have arisen by mutation of the genes coding for clavulanate-sensitive, plasmid-mediated beta-lactamases such as TEM-1, TEM-2 and SHV-1. Amino acid changes in ESBLs confer enhanced hydrolysis of oxyimino aminothiazolyl beta-lactams and aztreonam. Enzyme hyperproduction and loss of porins contribute to hydrolytic efficiency. ESBLs are highly susceptible to inhibition by clavulanate, and their presence can be detected by the disc approximation test, using amoxycillin/clavulanate and an ESBL-susceptible antibiotic. Other manual procedures have been used and commercial tests to detect the enzymes include Etest, Vitek and Dade Microscan products. The epidemiology of ESBLs is complex, and epidemic and sporadic strains may be encountered in the same hospital. Spread between hospitals--even countries--has been documented. ESBL activity is carried on large plasmids that often carry determinants for resistance to aminoglycosides and other antibiotics, and this is transmissible to Escherichia coli and other species of Enterobacteriaceae in which ESBLs have been detected. PMID- 11050777 TI - The bovine tubercle bacillus. AB - The bovine tubercle bacillus has always been eclipsed by the much higher incidence and the social and economic importance of its human cousin, as well as by the clinical unimportance of differentiating between the two. Nevertheless, in view of the resurgence of tuberculosis generally, the increase in the number of immunosuppressed individuals (i.e. the AIDS epidemic) and the 'great badger debate', there is renewed interest in it. Briefly, this review explores the history of Mycobacterium bovis and its potential for transmission between cattle and humans. PMID- 11050778 TI - Adherence mechanisms of Candida albicans. AB - The yeast Candida albicans is an opportunistic fungal pathogen that is capable of inducing a range of superficial and systemic diseases in the immunocompromised host. Although it displays a variety of virulence factors, one--the ability to adhere to host tissue--is considered essential in the early stages of colonisation and tissue invasion. Adherence is achieved by a combination of specific (ligand-receptor interactions) and non-specific (electrostatic charge, van der Waals forces) mechanisms which allow the yeast to attach to a wide range of tissue types and inanimate surfaces. Conventional methods for treating disease cause by C. albicans rely upon the use of antifungal drugs designed to kill the yeast or arrest its growth. An alternative approach, aimed at disrupting the adherence of the yeast to host tissue in cases of superficial infection, may have potential for controlling disease, particularly in situations where the unattached fungal cell can be removed from the affected site, either by the flushing action of the oropharynx or by the production of mucus in the vagina. PMID- 11050779 TI - Human research tissue banks in the UK National Health Service: laws, ethics, controls and constraints. AB - National Health Service (NHS) histopathology laboratories are ideally placed to bank cadaveric and surplus surgical tissue to supply the needs of research organisations, whether in the NHS, universities or the commercial sector. This review outlines the constraints to setting up a human tissue bank in an NHS histopathology laboratory, identifies the controls required and answers the question: is it ethical and legal? PMID- 11050780 TI - New perspectives on folate status: a differential role for the vitamin in cardiovascular disease, birth defects and other conditions. AB - In recent years, there has been heightened interest in the B vitamin folic acid, initially through its role in reducing neural tube defects, such as spina bifida, and, more recently, through its relationship with homocysteine and consequently the beneficial role it would seem to play in occlusive vascular disease. In addition, its sphere of influence may extend beyond these important conditions to include several cancers, Alzheimer's disease and affective disorders. The beneficial effects of folate in the above conditions can be explained largely within the context of folate-dependent pathways, such as methionine, purine and pyrimidine biosynthesis. However, the precise detail of folate metabolism is extremely complex and difficult to study because folate-dependent one-carbon metabolism is compartmentalised, involves an enormous number of low-abundance, difficult-to-measure, highly labile folyl coenzymes, and is the subject of genetic variability. Here we integrate some of the most recent findings in the field to provide a new perspective on folate status and some of the varied mechanisms by which folate ameliorates disease. PMID- 11050781 TI - Statins and the prevention of coronary heart disease: striking a balance that is desirable, affordable, and achievable. PMID- 11050782 TI - The primary prevention of coronary heart disease with statins: practice headache or public health? PMID- 11050783 TI - Management of UTI in general practice: a cost effective analysis. A commentary to facilitate an understanding of economic evaluation. PMID- 11050784 TI - The cost-effectiveness of lipid lowering in patients with ischaemic heart disease: an intervention and evaluation in primary care. AB - BACKGROUND: There has been a major revolution in the recommended treatment of hyperlipidaemia in patients with ischaemic heart disease following the publication of the Scandinavian Simvastatin Survival Study. This was the first major study to demonstrate that lipid-lowering drugs reduced mortality and morbidity in patients with ischaemic heart disease. AIM: To evaluate the feasibility and cost-effectiveness of screening and treating hyperlipidaemia in patients with ischaemic heart disease in primary care. METHOD: A study conducted in a rural dispensing training practice on the border of Nottinghamshire and Lincolnshire involving 327 patients with ischaemic heart disease who were registered with the practice on 1 January 1996. RESULTS: Eighty per cent of patients with ischaemic heart disease were considered eligible for screening and 80% of those attended for screening. The majority of patients who were screened had hyperlipidaemia that persisted after dietary advice. Despite lipid-lowering drugs, few patients had serum lipid concentrations in the target range at the end of six months. The costs of identifying and treating 83 patients with lipid lowering drugs over five years is estimated at 105,318 Pounds at 1996 prices, or 94,257 Pounds assuming a 6% discount rate per annum. Two-thirds of this is owing to the cost of lipid-lowering drugs. The discounted cost per coronary event prevented would be 17,138 Pounds (95% CI = 12,568 Pounds-26,183 Pounds). The discounted cost per coronary death prevented would be 32,502 Pounds (95% CI = 23,564 Pounds-55,445 Pounds). There were no important adverse effects of lipid lowering drugs on quality of life or mood. CONCLUSION: Such a programme is feasible and acceptable within primary care, although the ongoing cost implications need to be considered against the costs and benefits of other interventions. PMID- 11050785 TI - An assessment of morbidity registers for coronary heart disease in primary care. ASSIST (ASSessment of Implementation STrategy) trial collaborative group. AB - BACKGROUND: Organised care delivered systematically to all patients with established coronary heart disease (CHD) can reduce their risk factors and improve their quality of life. Therefore, identifying all patients with established CHD in a general practice population is an important first step for delivering this effective healthcare. However, there is little information on how registers are compiled, the factors that predict inclusion on the register or the relationship between registration and level of care provided. AIM: To assess the completeness of morbidity registers for CHD in primary care, the factors that predict inclusion on the register, and the relationship between registration and level of care provided. METHOD: Observational study at baseline of 1979 patients aged 55 to 75 years with established CHD in 18 general practices recruited for a cluster randomised controlled trial. RESULTS: The proportion of CHD patients correctly identified on practice morbidity registers varied from 29.3% to 100%. Four factors were significantly and independently associated with being on a register: a relevant surgery contact since diagnosis (OR = 2.1, 95% CI = 1.6% 2.9%); a relevant repeat prescription since diagnosis (OR = 1.6, 95% CI = 1.1% 2.3%); a diagnosis of myocardial infarction (OR = 1.5, 95% CI = 1.2%-1.9%); and a revascularisation procedure (OR = 1.5, 95% CI = 1.1%-2.0%). Inclusion on a register was strongly associated with being adequately assessed (i.e. assessed for smoking status, blood pressure, and cholesterol) (OR = 1.8, 95% CI = 1.3% 2.3%) and with treatment with aspirin or a lipid-lowering agent (OR = 1.4 for each agent). CONCLUSION: A wide variation in registration levels between practices exists. There is evidence that practices using multiple methods of case detection achieve higher levels of registration. The association between registration and better care does not prove causality but an effective call recall system is impossible without complete registration. PMID- 11050786 TI - Determinants of successful direct current cardioversion for atrial fibrillation and flutter: the importance of rapid referral. AB - Direct current (DC) cardioversion is an effective means of restoring sinus rhythm in patients with atrial fibrillation or flutter; however, the existing literature contains conflicting evidence on which factors are useful predictors of success. In a study of 171 patients undergoing DC cardioversion, we found that duration of arrhythmia prior to DC cardioversion was the only significant predictor of both successful cardioversion and subsequent maintenance of sinus rhythm (P < 0.001). Rapid DC cardioversion after the onset of atrial fibrillation or flutter significantly increases the likelihood of a successful outcome in both the short term and long-term. PMID- 11050787 TI - Observed changes in the lipid profile and calculated coronary risk in patients given dietary advice in primary care. AB - BACKGROUND: Dietary advice is usually the first-line treatment for increased blood cholesterol in primary care with a reduction in levels as the expected response. In practice, the diet adopted by the patient may lead to changes in blood lipids characterised by a greater decrease in high-density lipoprotein (HDL) than total cholesterol. The ratio of total cholesterol to HDL cholesterol is an important factor in calculated coronary risk using the Framingham model, from which most risk tables currently in use have been derived. This suggests that either coronary risk may increase after dietary advice or that risk should always be assessed on measurements made before any intervention has taken place. AIM: To report observed changes in blood lipids and calculated coronary risk following dietary advice in primary care. METHOD: Subjects with at least one coronary risk factor and baseline cholesterol above 5.2 mmol/l from an inner-city general practice had cardiovascular risk factors, including fasting lipids, recorded before receiving dietary advice. At follow-up several months later, risk factor measurements were repeated. Ten-year coronary risk was calculated using the Framingham model. Lipid levels and coronary risk at baseline and follow-up were compared. RESULTS: There was a significant decrease in both total cholesterol and HDL cholesterol in both sexes. However, in 56% of subjects, HDL decreased by a greater proportion than the total cholesterol. These subjects showed a highly significant increase in the total cholesterol/HDL cholesterol ratio (median = 0.8 [semi-interquartile range = 1.5], P < 0.001, which was correlated with a change in triglycerides (rs = 0.309, P < 0.001). In those who had an increase in the total cholesterol/HDL cholesterol ratio, calculated coronary risk increased from 5.45% (13.2) at baseline to 7.25% (15.5) (P < 0.001). In all subjects, the change in calculated coronary risk associated with dietary advice ranged from -15% to 15%. CONCLUSIONS: Low fat dietary advice in this primary care setting was frequently associated with undesirable changes in the lipid profile. The majority of subjects showed an increase in the total cholesterol/HDL cholesterol ratio, owing primarily to a decrease in HDL. Consequently, calculated coronary risk increased in over one-half of the subjects. Owing to our incomplete understanding of HDL metabolism, it is unclear whether the fall in HDL is actually detrimental; however, it seems prudent to give dietary advice to patients to avoid excess simple carbohydrate as a fat substitute. This helps avoid a rise in triglycerides, which appears to be associated with an increase in the ratio. These results confirm that coronary risk should always be calculated using measurements made before intervention. PMID- 11050788 TI - 'I always seem to be there'--a qualitative study of frequent attenders. AB - BACKGROUND: Much is still unknown about the consultation behaviour of frequent attenders, including why they consult as often as they do and why they consult in the patterns that they do. AIM: To determine why frequent attenders to general practice consult in the patterns that they do. METHOD: A qualitative study based on semi-structured interviews. Twenty-eight frequent attenders were purposively sampled from three practices; 13 exhibited a 'burst and gap' pattern of attendance and 15 exhibited a 'regular' pattern of attendance. RESULTS: A two part model is proposed. The first part encompasses each individual decision to consult and is based around eight questions that may be asked as part of the decision-making process (these concern the perception of the general practitioner's [GP's] role, past experience of symptoms and consulting, comparison with others' consulting, relationship with the GP, balancing fears, lay consulting, individual reasons, and whether it was a symptom that they would not normally consult for). The second part determines the pattern of consulting and has four major themes: predominantly medical reasons for attending, experience of what happens during the consultation, accessibility of the GP, and periods of not consulting. Two further themes are proposed: 'multiplicity', whereby the reasons for consulting lead to further consulting for related and unrelated problems, and 'passivity', whereby consulting seems to be out of control. CONCLUSIONS: The reasons underpinning each individual decision to consult were complex. The control that GPs were perceived to have over the pattern of consulting, for example concerning prescribing, review visits, and in addressing further help-seeking behaviour, may provide more possibilities for developing intervention strategies than targeting frequent attenders themselves. An understanding of the processes behind the consulting behaviour of frequent attenders may lead to more functional consultations and better clinical care as a result. PMID- 11050789 TI - A new approach to blood pressure measurement in the primary care setting. AB - In this study, a method of taking one blood pressure reading using a sphygmomanometer was compared with a method of taking multiple successive readings using an automatic device. With multiple readings the blood pressure tended to be lower and fewer patients were classified as hypertensive. Using an automatic blood pressure recording device seems to be a practical way of achieving multiple readings in a busy clinic setting. PMID- 11050790 TI - Atrial fibrillation: a comparison of methods to identify cases in general practice. AB - The importance of atrial fibrillation as a treatable risk factor for stroke is well established. Less is known about how to find previously unidentified cases within the community so that antithrombotic treatment can be offered to a wider group of at-risk patients. The aim of our study was to examine ways to improve the efficiency of practice-based screening for atrial fibrillation, including issues of time and financial cost. We used different combinations of pulse palpation and interpretation of 12-lead and bipolar electrocardiographs as carried out by practice nurses. The best strategy for the detection of atrial fibrillation in a practice population would appear to be to screen all eligible subjects by nurse pulse palpation, followed by 12-lead electrocardiograph readings in those who have a pulse suggestive of atrial fibrillation. The electrocardiograph interpretation can be undertaken effectively by a trained nurse. PMID- 11050791 TI - The work commitments of British general practitioners: a national survey. AB - Many qualified general practitioners (GPs) are choosing not to become principals. With the current problems in recruitment and retention of GPs, workforce planning for the future of general practice is contingent upon the work commitments of both GP principals and non-principals. A questionnaire survey of 5966 vocationally trained doctors in the UK suggests that shortfalls in the GP workforce will not be alleviated by relying on the non-principal pool increasing their time commitment to general practice work. PMID- 11050792 TI - Management of heart failure: evidence versus practice. Does current prescribing provide optimal treatment for heart failure patients? AB - Heart failure is an increasingly common and costly chronic disorder, with a rising prevalence of at least 2% in populations over the age of 45 years, mortality rates that are as poor as common solid cancers, and very high health care utilisation costs. Despite increased evidence supporting a range of effective interventions, predominantly therapeutic, there remain significant degrees of physician underperformance in terms of heart failure diagnosis and management. Until the early 1990s, the management of heart failure was largely confined to the symptomatic relief of patients with well established heart failure in fluid overload. The introduction of angiotensin-converting enzyme (ACE) inhibitors provided the first treatments that beneficially altered the prognosis of patients with the most common expression of heart failure, namely established systolic dysfunction, whether symptomatic or asymptomatic. Evidence has now extended these benefits to delaying progression of heart failure and reducing hospitalisation. Much of our understanding of the pathophysiology of heart failure stems from these studies. More recent data has clarified the limited role of digoxin, the important benefits of beta-blockade and aldosterone blockers as adjuvants to ACE inhibition, and the emerging evidence on angiotensin II antagonists. There are, in contrast to these positive findings, reliable data from Europe and North America revealing significant underperformance of primary care and hospital physicians in heart failure diagnosis and management, with evidence of underuse and underdosing of evidence-based therapies. Limited qualitative data suggest the reasons for this underperformance are complex and relate to lack of access to objective testing of ventricular function and exaggerated concerns over treatment risks and side-effects. Heart failure represents a complex cluster of aetiologies and risks that are not easy to correctly identify, even in specialist settings. Since there is now powerful evidence on how heart failure can be modified and improved, explicit guidance is needed for which suspected patients should be referred, for confirmation of diagnosis and advice on appropriate treatment regimes, and for which patients can be handled mainly within primary care but with enhanced access to objective non invasive tests to improve diagnostic reliability and to stratify patients to evidence-based therapies. Current evidence suggests that in North America and Europe today primary care physicians do underperform in their management of patients with heart failure, often owing to factors outside of their immediate control. PMID- 11050793 TI - The future general practitioner: out of date and running out of time. AB - In the late 1960s a Royal College of General Practitioners' working party produced a job description for the 'Future General Practitioner', together with an educational programme for vocational training. Despite the perceived success of vocational training, general practice remains academically disadvantaged compared with hospital medicine. Most general practitioners (GPs) have no contact with research or academic general practice, few achieve higher degrees compared with hospital consultants, and there are few academic posts in general practice. Junior doctors perceive general practice as offering less intrinsic job satisfaction than hospital medicine and recruitment is falling. Registrars who have completed vocational training are reluctant to commit themselves to general practice and often drift away from it. Schemes with an academic content, designed to retain doctors in general practice, have been well received but there are few career posts in academic general practice. Primary care groups and clinical governance will radically change the nature of general practice. GPs will no longer be at the centre of the primary health care team. Primary care trusts, serving populations of 100,000 or more at multiple sites, will still employ doctors but much of the traditional GP workload will be undertaken by nurses. Present day vocational training produces GPs without the skills that future 'community generalists' will need. Their training will be longer and their careers more structured than at present. They will use evidence-based practice routinely and be experts in information management, interpreting and managing complex diagnostic and therapeutic problems in the context of rapidly changing health technology. PMID- 11050794 TI - Questioning in general practice--a tool for change. AB - In general practice, as in all branches of medicine, doctors are encouraged to ensure their decisions reflect research findings, and are 'evidence-based'. This depends upon general practitioners (GPs) questioning their practice, finding 'evidence-based' answers, and changing their practice where necessary. Questioning behaviour is therefore fundamental to this process. Research into the questioning behaviour and information needs of GPs is difficult and it is unknown whether better access to information necessarily results in behavioural change or better health outcomes. This paper summarises research on doctors' questioning behaviour, factors influencing their likelihood of finding answers, and discusses some of the obstacles they face in implementing change. Finally, we introduce the concept of a 'clinical informaticist', whose role is to provide evidence-based answers to specific questions raised by GPs. This service may facilitate learning and increase uptake of research findings. PMID- 11050796 TI - Outpatient appointments. PMID- 11050795 TI - Importance of needle size for effective intramuscular delivery of vaccines. PMID- 11050797 TI - Counsellors in general practice. PMID- 11050798 TI - Psychological status and menstrual disturbance. PMID- 11050799 TI - Does excessive antibiotic use increase minor health complaints? PMID- 11050800 TI - Randomised trials in primary care. PMID- 11050801 TI - Managing constipation in children. AB - Constipation, defined as difficulty, delay or pain on defaecation, is common in children and is often difficult to manage. Here, we review the assessment and treatment of affected children and the support that they, and their families, may need. PMID- 11050802 TI - Managing the heavy drinker in primary care. AB - Estimates suggest that over 9 million adults in Britain drink alcohol at levels that could endanger their long-term health, and each year, in England and Wales alone, around 28,000 die prematurely from physical diseases, accidents and suicides related to alcohol use. The cost to the NHS of treating alcohol-related illness or injury may be as much as 400 million Pounds per year, and the wider costs to society are very much greater. GPs are well placed to identify patients who drink heavily. In this article, we discuss what part the primary healthcare team can play in the management of heavy drinkers. PMID- 11050803 TI - [Hantavirus pulmonary syndrome in southern Argentina]. AB - Andes virus was identified in 1995 as the etiologic agent of Hantavirus Pulmonary Syndrome (HPS) in Southern Argentina. We describe herein the main clinical characteristics of 25 HPS confirmed cases acquired in this area between 1993 and September 1999. The mean age was 34 years (range 11-70), with 72% males. Clinical characteristics were similar to those previously reported for Sin Nombre virus (SNV) cases. However, in this group of patients we also observed conjuntival injection in 10/25 (42%), facial flushing in 8/25 (33%), pharyngeal congestion in 7/25 (29%) and petechiae in 3/25 (12%). On the other hand, BUN was increased in 83% of cases (mean 0.77 g/l range 0.31-2.01). Mean serum creatinine concentration was 26.8 mg/l (range: 8.1-110 mg/l) with serum creatinine being higher than 20 mg/l in 8/15 patients (53%). Urinalysis was abnormal in 12/12 cases and was characterized by presence of proteins, red blood cells and granular casts. Aminotransferases were increased in 90% of cases with levels 5-10 times over normal values in 50% of cases. Serum creatine kinase concentration was elevated in 11/14 cases. Two patients required hemodialysis. Case fatality rate was 44% (11/25) and 10 of these cases died among the first 10 days of illness. Mononuclear myocarditis was observed in two cases, a finding that has not been reported for SNV cases. During the 1996 HPS outbreak in Southern Argentina due to Andes virus, there were epidemiological and molecular evidences of person-to person transmission, a feature not previously shown for other members of the hantavirus genus. These data would also be indicative of some distinctive clinical characteristics of HPS caused by Andes virus, with more frequent renal involvement than in SNV cases. PMID- 11050804 TI - [HIV-1 virus transmission through maternal milk]. AB - Human immunodeficiency virus type 1 (HIV-1) may be vertically transmitted during the pre, peri or postpartum period. Postnatal transmission as well as an increased risk of vertical transmission with breastfeeding has been shown for HIV 1 in several reports. Breastfeeding was here analyzed as a risk of HIV-1 transmission in a group of infants born to HIV-1 infected mothers. Among the 215 children studied in our population a significant difference was detected between those who were breastfed vs those who were bottle fed and finally became infected (p < 0.000000, R.R. = 4.29). We also report the case of a postnatal infection in a baby born to an HIV-1 seropositive father and a seronegative mother. Due to the risk of infection of the mother she had been thoroughly controlled when pregnant and after delivery. Mother and child were negative when retested at delivery, and at 10 months post-partum. At the age of 32 months the child attended the outpatient clinic with generalized lymphadenopathy and right parotitis. HIV-1 infection was then confirmed in both mother and child. At that time it was discovered that the baby had been breastfed up to the age of 24 months. This is the first reported child in Argentina whose infection may undoubtedly be attributed to breastfeeding. PMID- 11050805 TI - Bcl-2 molecular analysis in paraffin-embedded biopsies from diffuse large B-cell lymphomas. AB - Translocation t(14; 18) has been observed in 50-85% of follicular and in 30% of diffuse non-Hodgkin lymphomas. About half of follicle center lymphoma (FCL) undergo histological conversion at relapse to more aggressive diffuse large B cell lymphoma (DLBCL). This report correlates the molecular bcl-2/IgH rearrangement by PCR and Bcl-2 immunohistochemical (IHC) expression in a series of high grade DLBCLs with and without FCL remnant. Twenty-three paraffin-embedded lymph nodes from DLBCL patients were analyzed. Eleven patients showed FCL remnant (Group A) and 12, did not (Group B). Single PCR from paraffin extracted DNA followed by Southern transfer of products, hybridisation with internal oligoprobes for the MBR/JH and MCR/JH bcl-2 rearrangements and IHC analysis of Bcl-2 expression, were performed. PCR analysis was positive in 34.8% of patients. Bcl-2/IgH gene rearrangements were observed in 8 (34%) cases and 7 (30%) showed Bcl-2 expression on large noncleaved B-cells (centroblasts). All patients from Group A showed IHC positive reaction on FCL remnant (small cleaved cells) but only 2 (18%) were positive in DLBCL areas, suggesting either the loss of the bcl 2 expression on the transformed lymphoma, or, alternatively, the development of a second disease when the first lymphoma transforms. Group B patients showed a clear correlation between PCR and IHC studies. Our results suggest a similar frequency of t(14; 18) in DLBCLs to that reported in Europe and USA series. The discordance observed between PCR and IHC, particularly in Group A, points out the necessity to perform both studies in order to detect bcl-2 gene involvement in DLBCLs. PMID- 11050806 TI - Alkaline phosphatase isoenzymes for the diagnosis of metastatic tumors and lymphomas of liver and bone. AB - The plasmatic activities of total alkaline phosphatase (ALP) (E.C. 3.1.3.1), high molecular weight-ALP (high Mr-ALP) and bone-ALP isoenzymes, were determined in healthy individuals and in patients with: neoplasia without metastases, hepatic metastases, bone metastases and mixed metastases (hepatic and bone). Variables were individually used to assess incidence of metastases and percentages of false negative and false positive results were calculated. The three values were then used together to assess metastases incidence and sensitivity, specificity, positive predictive value, negative predictive value and predictive capacity were estimated. We conclude that none of the variables per se are reliable for the diagnosis of metastases. On the other hand, the three values show high percentages of sensitivity, specificity, positive predictive value, negative predictive value and a high probability (0.93) of accurate diagnosis when applied to a larger population, with similar prevalence values. PMID- 11050807 TI - [Chronic hereditary ataxic polyneuropathy]. AB - Sensory ataxic polyneuropathies are characterised by the presence of sensory ataxia due to damage to large myelinated sensory fibres, with total or relative preservation of muscle strength, pain and temperature sensation. Hereditary ataxic polyneuropathies are exceptional and very few families with this disorder have been reported so far. We here describe the neurological, electrophysiological and sural nerve biopsy data of four siblings with an ataxic chronic polyneuropathy, starting after age 50. They had an ataxic gait which worsened in darkness, horizontal nystagmus, hypo or areflexia, and severe impairment of limbs' propriocaption. Nerve conduction studies showed absent sensory nerve action potentials in all nerves tested. Somatosensory evoked potentials showed reduced amplitude and prolonged latencies. Sural nerve biopsy showed a severe loss of myelinated and unmyelinated fibres. Symptoms slowly progressed over the years. The recognition of this syndrome is important in the search for the etiology of chronic ataxic neuropathies. PMID- 11050808 TI - [Amantadine for the treatment of levodopa dyskinesias in Parkinson's disease]. AB - The development of dyskinesias is a common side effect during chronic levodopa therapy in parkinsonian patients. Recent reports suggest that amantadine, a drug with well known antiparkinsonian activity, is effective in the treatment of this complication. In order to evaluate its usefulness we conducted an open label, prospective and longitudinal study in 26 patients with Parkinson's disease (PD) on chronic levodopa therapy who presented peaks of dose dyskinesias. After 3 weeks' treatment dyskinesia severity was reduced by 70% (p < 0.0001) on the I SAPD scale and by 68.8% (p < 0.0002) on the UPDRS IV subscale. Patients were later evaluated every 45 days showing persistent drug benefit during follow-up ranging from 2 to 11 months (mean 6.5 months). One third of our series presented unwanted effects which were only severe enough in 2 cases to discontinue treatment. In the others, side effects were transient or readily abated with amantadine dose reduction. Our findings support amantadine as a safe and useful drug to treat levodopa-induced dyskinesias which on occasion prove as disabling as PD itself. Treatment with amantadine should routinely be considered before indicating pallidotomy for levodopa-induced dyskinesias. PMID- 11050809 TI - [Detection of preclinical Cushing's syndrome in overweight type 2 diabetic patients]. AB - Preclinical Cushing's syndrome (PCS) is a condition in which cortisol excess is not associated to clinical features of Cushing's syndrome. The aim of this study was to detect PCS in 48 ambulatory overweight type 2 diabetic patients (DM). Controls were 40 normoglycemic obese (Ob) and 36 normo-weight healthy subjects (N). In DM (47/48) total urinary cortisol (UF) levels were similar to those found in Ob and N. Evening urinary cortisol (Spot F) was significantly higher than either Ob (p: 0.0001) or N (p: 0.03), although values did not overcome the upper normal limit (44 ng/mg creatinine). False positive results to the overnight 1 mg dexamethasone suppression test were found in 31% and 22% of DM and Ob, respectively. In a DM female an elevated UF and Spot F associated to absence of cortisol inhibition to the overnight 1 mg dexamethasone suppression test was repeatedly detected. Diagnosis of PCS was performed. Remission of hypercorticism and glycemic control were achieved after pituitary surgery. It would be useful to screen DM patients with poor glycemic control for PCS. PMID- 11050810 TI - Assessment in the diagnosis of male chronic genital tract infection. AB - Different methodologies have been proposed to interpret the microbiological findings associated with contaminating, indigenous microbiota of the anterior urethra. In order to solve the controversy related to the diagnosis of chronic seminal infections in asymptomatic young adults, the results applying Stamey and Meares' criteria were compared with those obtained when semen cultures were studied for significant bacteriospermia. A total of 218 consecutive asymptomatic male partners of infertile couples were evaluated by the four-specimen technique described by Stamey and Meares' with the addition of semen (SM). Infection was detected in 46% by SM, while semen cultures (SC) showed a prevalence of infection of 41%; 73 patients were positive by both criteria and 102 negative; 27 patients were positive by SM technique in prostate fluid while their semen cultures were negative; 16 patients had positive semen cultures and were considered negative by SM. The kappa statistic indicated a good degree of agreement between both methodologies (kappa = 0.61, z = 8.68, p < 0.001). The estimated risk of being considered negative attributable to the semen culture (27 patients) was 25% (attributable risk = gamma ac- = 0.2550), and of being considered positive attributable to the semen culture (16 patients) was 26% (gamma ac+ = 0.2579). The 95% confidence limits were estimated in 12 to 39%, and in 13 to 31%, respectively. In view of these results, to establish the diagnosis of chronic prostatitis, the addition of prostatic fluid or voided urine cultures after prostatic massage, must be performed. Semen culture confronted with first-voided urine avoid overestimating seminal infection. PMID- 11050811 TI - [Acute confusion syndrome in the hospitalized elderly]. AB - Our purpose was to determine the in-hospital incidence of delirium among elderly patients, its relation to previous cognitive impairment and the time between admission and its development. We performed an observational study of follow-up in the internal medicine area of a university hospital. We included consecutively and prospectively every patient 70 years or older upon admission. Patients with delirium on admission were excluded, as also were those taking antipsychotic drugs, with severe language or audition impairment, or coming from other sites of internation. We subsequently eliminated patients whose follow-up had not ended by the time the study was concluded, and patients in whom psychosis was diagnosed. Clinical and laboratory data were collected, and patients were prospectively followed until discharge from the hospital, using the Confusion-Assessment-Method (CAM) for the diagnosis of delirium. We analyzed 61 patients of whom 13 developed delirium while hospitalized (in-hospital incidence: 21.31%--CI 95%: 11.03 31.59%). Patients with delirium had had lower scores on Mini Mental State upon admission (median 17 vs 22; p 0.001). During the first 4 days of hospitalization 58.3% of delirium cases occurred not modifying the duration of hospitalization (average: 10.22 days vs 14.38; p = NS). We conclude that the incidence of delirium is high among hospitalized elderly patients specially during the first days, and in those with previous cognitive impairment. We suggest that delirium could be an associated disorder in severe diseases among patients with previous cognitive damage. PMID- 11050812 TI - Circulating and mitogen-induced tumor necrosis factor (TNF) in malnourished children. AB - Malnutrition in children is associated with an increased risk of infection and death. Multiple abnormalities in the inflammatory-immune response, including cytokine production, have been described in protein energy malnourished (PEM) children and could account for increased severity and frequency of infection. The aim of the present study was to determine whether there are abnormal basal tumor necrosis factor (TNF) serum concentrations in PEM children, to relate it with serum cortisol and plasma corticotrophin levels and to explore simultaneously the in vitro production of TNF by peripheral blood leukocytes (PBL). No differences were found in basal plasma corticotrophin and serum cortisol concentrations in malnourished as compared with normal, well-nourished control children. Basal TNF serum concentrations were significantly higher in malnourished children than in controls. Conversely, mitogen induced TNF production by PBL in vitro was significantly reduced in PEM children compared with controls. Abnormalities in circulating and mitogen-induced TNF production are present in malnourished children even in absence of elevated serum cortisol concentrations. These abnormalities potentially could modify inflammatory-immune responses to infectious stimuli in malnourished children. PMID- 11050813 TI - [Syphilis and pregnancy. Prenatal control, seroprevalence and false biological positives]. AB - Syphilis may be transmitted vertically, especially if the mother is in an early stage with a high bloodstream treponema concentration, although it may also be transmitted to a lesser degree in late latency, when non-treponemic serology may become negative spontaneously with persistence of treponemic serology. The prenatal control for syphilis is routinely carried out by means of a non treponemic reaction such as VDRL or rapid plasma reagin (RPR) which, when positive, should be confirmed by treponemic techniques such as fluorescent treponemal antibody absorption (FTA-abs) and/or hemagglutination (MHA-Tp). Prevalence of syphilis should be defined on the basis of positive treponemic reactions. To define the seroprevalence and the validity of these control guidelines, 1,056 pregnant women attending the Hospital de Clinicas for their initial control were evaluated by means of serological treponemic and non treponemic methods. Serological results disclosed 4 distinct groups. Group 1 (n = 17 or 1.61%) presented both types of reactive tests, while Group II (n = 22 or 2.08%) only presented reactive treponemic tests, and both groups were seroreactive for syphilis. Group III (n = 7 or 0.66%) only showed reactive non treponemic tests, which were considered biological false-positive (BFP) reactions. Five of them were reactive for antiphospholipid antibodies. Group IV (n = 1,010 or 95.65%) fell to present serological evidence of syphilis. To conclude: 1) global seroprevalence in this population was 3.69%; 2) since 2.08% of pregnant seroreactive mothers had not been detected by routine screening, it would be advisable to perform simultaneous treponemic and non-treponemic techniques for prenatal control. 3) This methodology should identify the BFP tests in the same screening. PMID- 11050814 TI - [Prevalence of Flavivirus antibodies in Alouatta caraya primate autochthonous of Argentina]. AB - Flavivirus constitute a human health problem in our country. Primates are known to participate in the maintenance of Dengue and Yellow Fever viruses. However, these animals play a role which still remains to be determined in the maintenance of other viruses with potential pathogenicity for human beings and/or animals. Deteccion of antibodies was performed for different flavivirus in 105 sera samples of Alouatta caraya primates by the hemagglutination inhibition (HI) test. The neutralization (NT) test confirmed only infections caused by St. Louis Encephalitis (SLE) virus with a high prevalence in HI (35.23%) and NT (32.38%) antibodies. No antibody titres indicative of infections by Yellow Fever, Dengue and Bussuquara viruses were registered. Infection by the liheus virus could not be confirmed in these primates. There is a need for studies to detect new or reemergent viral infections in Argentina and the role that these primates could play in the maintenance of such infections. PMID- 11050815 TI - [Cavum lymphoma in a hemophilic patient with AIDS]. AB - Intermediate and highly malignant non-Hodgkin and primary central nervous system lymphomas are marker diseases for AIDS. Cavum and oropharynx involvement by these tumors is uncommon. Although there are few cases reported in the literature, these may be primary localizations of the tumor. We present a hemophilic HIV+ patient with non-Hodgkin lymphoma of the cavum. The histologic diagnosis was high grade, pleomorphic, centroblastic lymphoma. The patient was treated with chemotherapy plus intrathecal chemotherapy and highly active antiretroviral therapy (HAART). His evolution has been excellent. One year after diagnosis, the patient is asymptomatic with no evidence of residual tumor, and responding well to HAART. PMID- 11050816 TI - [Diagnosis by electron microscopy of recessive dystrophic epidermolysis bullosa]. AB - Inherited epidermolysis bullosa (EB) includes a number of distinctive diseases that are characterized by the presence of fragile skin and the tendency to develop blisters and erosions. The current classification separates the types of EB on the basis of the ultrastructural level of the blisters. The electron microscopy is very important for the diagnosis and in the recessive dystrophic EB shows that the lamina densa forms the roof of the blister and that the number of anchoring fibrils are absent or reduced. We present the case of a 30 year old woman with a diagnosis of recessive dystrophic EB diagnosed by electron microscopy. PMID- 11050817 TI - [Coxitis due to multidrug resistant Mycobacterium tuberculosis in a HIV negative patient]. AB - A case of an HIV negative female patient with coxofemoral arthritis of tuberculous etiology, multidrug-resistant strain, and connective tissue disease associated to glucocorticoid therapy is reported. The patient was treated with cycloserine, ethambutol, p-aminosalicylic acid and ofloxacin, with improvement of the joint lesions. Previous publications on this subject are reviewed. PMID- 11050818 TI - [Bilateral and simultaneous cerebellar infarct]. PMID- 11050819 TI - [Aldosterone-producing adrenal adenoma]. PMID- 11050820 TI - [Hypertension, diabetes, hemiparesis and sepsis in an elderly woman]. PMID- 11050821 TI - [Guidelines for the treatment of aneurysmal subarachnoid hemorrhage. A consensus conference]. AB - The subarachnoid hemorrhage (SAH) by aneurysmatic rupture is responsible for 6% of the cerebral vascular accidents. The cerebral aneurysms are present in 0.2 9.9% of the population and the bleeding rate is of 10 out of 100,000 inhabitants per year. The consensus conference analyzed the different schemes of treatment and made therapeutic recommendations according to the criteria of medicine based on evidence. Levels of evidence were determined from I to V. The recommendation degrees were classified in: A, determined by evidence level I, B by evidence level II, and C suggested by evidence levels III, IV and V. These recommendations should be adapted to each patient. However, grade A recommendations are treatment standards. Seriousness of patients was evaluated on the basis of Hunt and Hess scale upon admission. Successive analyses covered: general medical treatment measures, cerebral vasospasm, diagnostic procedures and treatment of the hyponatremia and convulsion prevention. PMID- 11050822 TI - [Programmed cell death and apoptosis. The role of mitochondria]. AB - Physiological cell death and apoptosis are natural processes genetically programmed, subjected to control by complex molecular mechanisms which elucidation is of particular interest for biology and medicine. Mitochondria play an essential role in physiological cell death and apoptosis. Apoptogenic effects develop in three phases, namely: (a) premitochondrial; (b) mitochondrial and (c) post-mitochondrial. During the first phase, apoptogenic signals (genotoxic agents, oxygen free radicals, corticoids, antibodies, etc.) interact with cell receptors activating specific mechanisms including thiol dependent proteases (caspases). As a consequence of those signals, mitochondrial damage results (membrane permeabilization, collapse of the membrane potential, swelling, membrane disruption, inhibition of electron transfer and oxidative phosphorylation). Other consequences of the mitochondrial disruption are the enhancement of free radical production and the exit of cytochrome c, caspases and endonucleases to the cytosol. During the third phase of apoptosis, free radicals and activated enzymes attack the cell protein structure and ADN, thus causing cell death. The mitochondrial regulation of apoptosis is controlled by the mitochondrial transitory permeability pore (MTPP) which is constituted by caspases, hexokinases, cytochrome c, ATP and ADP. MTPP is subjected to control by apoptogenic or antiapoptogenic agents which open or close it, according to their structure and the cell metabolic conditions. Uncontrolled opening of MTPP determines a massive exit of mitochondrial apoptogenic factors which in the cytosol and the nucleus exert their apoptogenic effects, thus producing cell death. MTPP can be modified by drugs with potential therapeutic actions thus opening interesting therapeutic possibilities. The role of apoptosis in pathologies such as degenerative diseases of the nervous system, autoimmunity diseases, SIDA and cancer is discussed. PMID- 11050823 TI - [Spontaneity and the cardiac pacemaker]. PMID- 11050824 TI - [Women in science. A personal vision involving 60 years of research]. PMID- 11050825 TI - [New therapeutic approaches to sepsis. Illusions, failures and errors]. PMID- 11050826 TI - [Competitiveness in science. Today, tomorrow and for ever]. PMID- 11050827 TI - [Evolution of a patient with a partial deficiency of lecithin:cholesterol acyltransferase]. PMID- 11050828 TI - [Campylobacter jejuni bacteremia in a patient with non Hodgkin lymphoma and pneumonia]. PMID- 11050829 TI - [Capacitation of residents in women's health]. PMID- 11050830 TI - [Psychiatric diagnosis: label, disorder, comorbidity]. AB - This editorial emphasizes the descriptive character of two of the most widely employed psychiatric diagnostic systems currently in use: DSM-IV and ICD-10. While they cannot be deemed atheoretical, they strive for neutrality and propose a nomenclature devoid of derogatory connotations. It is contended that the lowered "labeling threshold" of these systems tends to identify conditions that, while abnormal in the sense of producing personal discomfort or dysfunction, could not always be considered diseases requiring intervention from healthcare systems. This distinction is useful when evaluating studies which acritically apply screening instruments with a diagnostic purpose. These studies, while producing data, may not always be relevant for empirical studies of comorbidity or prevalence. PMID- 11050831 TI - [Infective endocarditis: short and long-term results in 261 cases managed by a multidisciplinary approach]. AB - BACKGROUND: Early diagnosis, an effective treatment and prompt recognition of complications are essential to improve the prognosis of infective endocarditis (IE). AIM: To report the results of a multidisciplinary approach to diagnosis and management of patients with IE at the Universidad Catolica de Chile Hospital. PATIENTS AND METHODS: The clinical history, diagnosis, treatment and outcome of 261 episodes (Duke criteria) of IE admitted between January 1980 and January 1999 were analyzed. These included 185 episodes of native, 73 of prosthetic valve and 3 of nonvalvular IE. RESULTS: Sixty nine percent of patients were men and the mean age was 49 +/- 16 years. Seventy five percent had a definite diagnosis of IE (Duke). S. viridans, staphylococci and enterococci together constituted 85% of the isolated bacterial strains. Twenty seven had culture-negative IE, related to a high incidence of antibiotic therapy prior to diagnosis. Transesophageal echocardiography was performed in 102 cases and it detected vegetations in 91% of aortic and 96% of mitral IE, rupture or prosthesis dehiscence in 67% of aortic and 52% of mitral IE and abscesses in 51% of aortic and 15% of mitral IE. Fifty one percent developed heart failure and 34% had embolic events. S. aureus IE was associated to a higher incidence of embolic events, complications which contraindicated surgery and increased mortality rate (27%). Of all patients, 40% were treated exclusively with antibiotics, 52% were operated on and 8% had surgical indication but were nonoperable because of serious complications. The overall mortality was 16.3%: 13% in the medical, 9% in the surgical and 81% in the non-operable groups. The type of treatment and mortality rates did not differ between IE of native valves and prosthetic valves. Long term follow up showed survival rates of 73% at 5 years and 66% at 10 years. CONCLUSION: A multidisciplinary approach may be very helpful to improve the prognosis of IE. PMID- 11050832 TI - [Study of association between the magnitude of sphincter hypotonia and esophageal motor disorders]. AB - BACKGROUND: One of the most important factors involved in the pathophysiology of gastroesophageal reflux disease (GERD) is the lower esophageal sphincter rest pressure (LESRP), but these patients can have esophageal motor disorders (EMD). AIM: To assess an association between LESRP and the appearance of EMD in patients with GERD. SUBJECTS AND METHODS: A cross-sectional study was conducted in 229 patients with GERD and 49 healthy controls. Forty five patients with LESRP < 6 mmHg and a mean age of 49 years were assigned to group 1, 128 patients with a LESRP between 6 and 12 mmHg and mean age of 47 years were assigned to group 2, 56 patients with a a LESRP > 12 mmHg and a mean age of 47 years were assigned to group 3 and group 4 was conformed by 49 healthy subjects aged 40 years old. Esophageal manometry was performed using previously published techniques. RESULTS: There was a significant association between LESRP, waves amplitude and the frequency of tertiary waves. CONCLUSIONS: Resting lower esophageal sphincter pressure is inversely proportional to the presence of esophageal motor disorders in patients with gastroesophageal reflux disease. PMID- 11050833 TI - [Comorbidity of chronic diseases and psychiatric disorders among patients attending public primary care]. AB - BACKGROUND: As a part of the World Health Organization multicentric study of emotional disorders in general medical care, we studied patients who had a chronic medical ailment and a psychiatric disorder, according to ICD-10. AIM: To report the prevalence of patients with coexisting medical and psychiatric disorders. PATIENTS AND METHODS: All patients, aged 15 to 65 years old, consulting in primary care outpatient clinics, were interviewed using a general health questionnaire. In a second phase, patients with chronic medical disorders were subjected to the World Health Organization Composite International Diagnostic Instrument. RESULTS: Sixty nine percent of interviewed Chilean patients had a medical condition, compared to 60.3% of the global study group. Of these, 66% had a coexisting psychiatric diagnosis, compared to 31% of the global study group. The most frequent diagnoses in the Chilean sample were somatization disorders in 25%, harmful alcohol use in 14%, depression in 35% and hypochondriasis in 6%. There was a higher prevalence and odds ratio for psychiatric diagnoses among Chilean women. CONCLUSIONS: Patients with chronic medical disorders should be considered a high risk group for the coexistence of psychiatric disturbances. PMID- 11050834 TI - [Prevalence of anti-hantavirus antibodies in health care personnel in direct contact with patients with hantavirus pulmonary syndrome in Temuco, Chile 1997 to 1999]. AB - BACKGROUND: An outbreak of Hantavirus Cardiopulmonary Syndrome (HVCS) caused by the Andes virus, affected Chile since 1995. Antibodies to Hantavirus in health care workers who had cared patients with HVCS in Coybaique, Argentinean reports and familial clustering of bantaviral illness, raised the possibility of person to person transmission. Familial clustering could occur secondary to a similar exposure to a common infected environment of more than one member of the family. Moreover, the prevalence of antibodies in health care workers in Coyhaique does not differ from the prevalence in general population in that region. AIM: To study the prevalence of antibodies to Hantavirus in health care workers exposed to body fluids of 20 patients. MATERIAL AND METHODS: Among health care workers exposed to patients with HVCS, we registered information about the exposure to patients and to the environment outside the hospital in which they could have been infected. IgG antibodies against Hantavirus were measured by ELISA using two dilutions. RESULTS: Sixty seven workers were studied. Of these, 73% were exposed to respiratory secretions and blood, 21% to blood and 6% to respiratory secretions. Only 6% protected themselves properly, 49% used facial masks and gloves, 25% only facial masks, 7% only gloves and 12% used no protection measures. In none of these workers, Hantavirus antibodies were detected. CONCLUSIONS: These results are supporting evidence against person to person transmission of the Andes virus. PMID- 11050835 TI - [Lipid profile in newborns with intrauterine growth retardation]. AB - BACKGROUND: The X syndrome, related to coronary disease in adults, could be possibly programmed priory to delivery, in children with intrauterine growth retardation. AIM: To measure serum lipids in newborns with symmetrical or asymmetrical intrauterine growth retardation. PATIENTS AND METHODS: One hundred thirty-five newborns with intrauterine growth retardation and 116 normal term newborns, with 38 to 41 gestational weeks, were studied. Total, HDL, and LDL cholesterol, triglycerides and apoproteins. A1 and B were measured in umbilical cord blood samples. RESULTS: No differences in total, HDL, LDL cholesterol, apoproteins A1 and B were observed between the study groups. Triglycerides were higher in newborns with intrauterine growth retardation, compared to normal term newborns (45 +/- 27 and 36 +/- 19 mg/dl respectively, p < 0.001). Differences in serum triglyceride levels respect to controls were observed in both male and female newborns with asymmetrical growth retardation. Likewise the differences respect to controls were observed in newborns with mild or severe but not with moderate growth retardation. CONCLUSIONS: Newborns with intrauterine growth retardation have higher triglyceride levels than normal term newborns. PMID- 11050836 TI - [Prospective, randomized, comparative study of the efficacy, safety and cost of cefuroxime versus cephradine in acute pyelonephritis during pregnancy]. AB - BACKGROUND: Second generation cephalosporins (CFPs) are more active in the treatment of acute pyelonephritis during pregnancy but their cost is considerably higher than their predecessors. Cefuroxime, a second generation CFP with oral and parenteral presentations, might offer significant advantages and become a first choice antimicrobial in this setting. AIM: To compare the efficacy, safety and cost of cefuroxime and cephradine in the treatment of acute pyelonephritis in pregnancy. PATIENTS AND METHODS: Hospitalized women with 12 to 34 weeks of pregnancy, with clinical and bacteriological diagnosis of acute pyelonephritis, were randomly assigned to receive cefuroxime (Curocef(r), Glaxo Wellcome) 750 mg t.i.d, i.v. or cephradine 1 g q.i.d., i.v. If the isolated organism was resistant to the assigned drug the patient was excluded. Once patients were afebrile, they were switched to an oral form of the same antimicrobial. They were discharged according to the clinical status and treated for a total of 14 days. laboratory tests, including urine culture were requested during controls and at the end of follow-up at 28 days. RESULTS: One hundred and one patients were randomized: 49 to receive cephradine and 52 to receive cefuroxime. Patients in the cefuroxime group hed fewer febrile days (mean 1.7 vs 2.2, p < 0.05), faster clinical recovery (mean 2.7 vs 3.1 days, p < 0.05), a higher rate of bacteriological cure at 28 days (78.8% and 59.2%, p < 0.05) and lower rate of failure (21.2% vs 40.8% p < 0.05). The rate of resistance of isolated uropathogens was 14% to cephradine and 1% to cefuroxime. CONCLUSIONS: Cefuroxime can be considered as a first choice option in the treatment of acute pyelonephritis during pregnancy due to its tolerance, microbiological activity and efficacy. PMID- 11050837 TI - [Prevalence of the Chlamydia trachomatis in neonatal conjunctivitis determination by indirect fluorescence and gene amplification]. AB - BACKGROUND: Chlamydia trachomatis is one of the most common identifiable infectious agents in neonatal conjunctivitis. It also causes pneumonitis, that is preceded by conjunctivitis in one third of cases. AIM: To asses the prevalence of Chlamydia trachomatis in newborns with conjunctivitis. PATIENTS AND METHODS: In 162 newborns, coming from 14 Primary Health Centers from Santiago de Chile, C. trachomatis was detected by indirect fluorescence and two polymerase chain reaction (PCR 1 and 2), which amplified different sequences from the common endogenous plasmid. Those patients with positive indirect fluorescence and PCR 2 were defined as infected: RESULTS: The prevalence of C. trachomatis was 8%, and the distribution of the positive cases was similar in the different Health Centers. Other isolates were: S. aureus (9.8%), S. pneumoniae (8%), S. viridans (6.2%) y H. influenzae (5.5%). CONCLUSIONS: The prevalence of C. trachomatis in neonatal conjunctivitis in Chile is similar to that of developed countries. Therefore, C. trachomatis should be considered in the election of antimicrobials for the treatment of neonatal conjunctivitis, to avoid ocular and respiratory complications. PMID- 11050838 TI - [Evaluation of Nugent and Amsel criteria for the diagnosis of bacterial vaginosis]. AB - BACKGROUND: Bacterial vaginosis (BV) is a common disease in reproductive-age women and is associated to important gynecologic and obstetric complications. AIM: To study the occurrence of BV in apparently healthy women attending family planning clinics, using Amsel and Nugent diagnostic criteria. MATERIAL AND METHODS: Two hundred thirty nine women consulting for symptoms associated to cervicovaginitis, were studied. A sample from the lateral walls of the vagina was obtained with a sterile swab for microscopic analysis, Gram stain and amine test. RESULTS: According to Amsel and Nugent criteria a 31.1% and 31.8% BV prevalence was observed. The sensitivity and specificity of Nugent criteria, compared with Amsel criteria were 83.3% and 92.1%, respectively. CONCLUSIONS: The high prevalence of BV found in this study suggests that this vaginal infection should be diagnosed with standardized methods. Nugent criteria are economic easy to perform and sensitive and we propose that they should be used in local health centers. PMID- 11050839 TI - [Post exercise myalgias as presentation form of dystrophinopathy]. AB - Cramps and myalgias are frequent presentations of many disorders whose diagnosis is generally difficult. Among the unusual causes stand the milder phenotypes of dystrophinopathies, which are caused, just as Duchenne and Becker's dystrophy, by mutations in the dystrophin gene. An 8 year-old boy presented severe muscle pain on exercise and serum rise in creatine kinase over 1000 U/l. He had normal muscle power and mild calf hypertrophy. The molecular analysis by polymerase chain reaction (PCR) of the dystrophin gene showed deletions of exons 45 to 51. Dystrophin analysis by Western blot revealed a dystrophin of reduced quantity and molecular weight. Emphasis is made to include dystrophinopathies in the differential diagnosis of myalgias and the usefulness of molecular genetic techniques in the identification of these disorders. PMID- 11050840 TI - [Microdeletion of Y chromosome in severe olygozoospemic infertile patient]. AB - We are reporting a 37 year old male with severe oligozoospermia and a history of infertility for thirteen years and surgery for severe unilateral varicocele. The hormonal levels for FSH, LH and T, and karyotype were within the normal range. Multiplex PCR revealed the presence of a de novo microdeletion in the azoospermia factor (AZF) c region involving the deleted in azoospermia (DAZ) and basic protein Y-2 (BPY2) genes. These results suggest that severe oligozoospermia should be considered for the screening of microdeletions of Yq involving the AZFc region even in the presence of a varicocele. PMID- 11050842 TI - [Reflections about the book "El fin de la medicine" of Alejandro Goic] [In Process Citation] PMID- 11050841 TI - [Thyroid lymphoma. A case report]. AB - Thyroid lymphoma represents less than 1% of malignant thyroid tumors and its diagnosis is difficult. We report a 25 years old woman, admitted with the diagnosis of diffuse euthyroid goiter and thyroid cancer. She was subjected to a subtotal thyroidectomy and the pathological study of the surgical piece showed a Hodgkin lymphoma, subtype nodular sclerosis. The patient was treated with three cycles of chemotherapy, using cyclophosphamide, vincristine, procarbazine, prednisone, doxorubicin, blemycin, vinblastin and radiotherapy. She refused to continue treatment after the third cycle and after 3 years and 5 months of follow up, is well and free of disease. PMID- 11050843 TI - [Multiple endocrine neoplasia: a clinical model for applying molecular genetic techniques]. AB - Multiple endocrine neoplasias (MEN) are syndromes inherited as autosomal dominant. The application of the techniques of molecular biology has made possible the identification of the genes causing MEN 1 and 2. The gene responsible for MEN 1 belongs to the family of tumor suppressor genes and encodes for a protein named MENIN whose function remains to be elucidated. The identification of mutant MEN 1 gene carriers who are at risk of developing this syndrome requires frequent biochemical screening for the development of endocrine tumors. MEN 2 is a consequence of mutations in the Ret proto-oncogene (c-Ret). This gene encodes for a tyrosine kinase receptor thought to play a role in the development of neural crest-derived tissue. Members of kindred with either MEN 2A or MEN 2B should be screened by direct DNA testing early in life for mutations in c-Ret. Those with the mutation should be advised to have thyroidectomy at five years of age in children with MEN 2A and earlier in children with MEN 2B. Some cases of sporadic MTC are actually MEN 2A or Familial MTC after c-Ret testing is done, therefore routine application of this test is recommended in all cases of apparent sporadic MTC. PMID- 11050844 TI - [The Beca's in Chilean psychiatry]. AB - The Chilean psychiatrists, Manuel and Francisco Beca, father and son, who lived at the turn of the XIX century and in the first half of the XX respectively, dedicated their lives to the care of mental patients and to the teaching of psychiatry, contributing in their own way and time to the development of the specialty in Chile. Manuel Beca, a clinician, published the first mental patient statistics and Francisco, an academic, became professor of Psychiatry at the Catholic University. Although they dedicated themselves to different fields in the area of mental health, such as clinical research and teaching, both have outstanding merits to become a part of the historical memory of Chilean psychiatry. PMID- 11050845 TI - [Definitions in medicine: do ends justify the means and medical actions?]. PMID- 11050846 TI - [Situation of osteoporosis in Chile]. PMID- 11050847 TI - The consent of the governed. PMID- 11050848 TI - Qualitative analysis: a guide to best practice--forensic science extension. AB - Qualitative analysis is arguably the cornerstone of analysis since what cannot be recognised cannot be quantified. This is especially true in the case of the forensic sciences where, for example, the mere presence of a prohibited, or controlled, substance in an unauthorized setting is indicative of a potential offence. This paper presents a set of principles of good practice in qualitative analysis. The principles are placed in the context of forensic science and extend earlier published guidance. PMID- 11050849 TI - The potency of cannabis in New Zealand from 1976 to 1996. AB - The potency of cannabis plant and cannabis products seized in New Zealand over the period of 20 years is studied. The earlier part of the study includes mainly imported cannabis oil and cannabis resin, and both imported and locally grown cannabis plant, that was seized by the police. The later part of the study includes little imported material. Cannabis plant is now locally grown, cannabis oil is locally manufactured and imported cannabis resin is rarely seized. The average potency of the cannabis plant available to the user has not increased over the 20 years period. Cannabis leaf contains on average 1% THC and the female flowering heads on average 3.5% THC. The average potency of cannabis oil has dropped from its peak at 34% THC in 1985 to 13% THC in 1995. PMID- 11050850 TI - eHealth 2000/TEPR focus on patient empowerment. PMID- 11050851 TI - Using the Web as a research tool. PMID- 11050852 TI - Public health, clinical data, and common cause: information standards as mediating foci. PMID- 11050853 TI - Ambulatory payment classifications: information technology to the rescue. PMID- 11050854 TI - Laboratory of computer science, Massachusetts General Hospital: four decades of medical informatics. PMID- 11050855 TI - Partners HealthCare. Creating and managing an integrated delivery system. AB - Supporting the "integration" of an integrated delivery system is an exceptionally complex and difficult challenge. Lack of organizational clarity about integration strategies and value, political challenges in effecting integration, the idiosyncratic and evolutionary nature of integration, and the technical challenges of integrating heterogeneous technologies all contribute to the scale of the challenge. Information systems integration strategies and tactics should be guided by overall concepts that frame the organization's understanding of the nature of integration. Effort must be directed to working with IDS leadership to define the approach to specific classes of integration, e.g., clinical integration. And, as always, one should learn from the experiences of others. While it may be difficult to fully define the value of integration in an IDS, the IDS seems to be a permanent feature of the healthcare landscape. And the integration of the IDS has value, value that is often nuanced, intangible, and complicated. Hence, the efficient and effective application of information systems to further integration is a critical organizational undertaking. PMID- 11050856 TI - Beyond the double doors. New generation surgical information systems. PMID- 11050857 TI - Computer-assisted orthopedic surgery. Tools and technologies in clinical practice. PMID- 11050858 TI - Public key infrastructure. Securing the exchange of health information. PMID- 11050859 TI - The NRC report. Is healthcare ready for the Internet? PMID- 11050860 TI - The use of a registered trade mark. Velcro Group Corporation. PMID- 11050861 TI - Tactical management of urban warfare casualties in special operations. PMID- 11050862 TI - Personality characteristics that increase vulnerability to sexual harassment among U.S. Army soldiers. AB - The study examines personality characteristics that may increase vulnerability to sexual harassment among active duty Army personnel. A survey was administered to 1,060 male soldiers and 305 female soldiers at three Army posts located in the United States. Sexual harassment was measured by the Sexual Experiences Questionnaire, which assesses unwanted sexual behaviors in the workplace. Two gender-related personality characteristics derived from the Extended Personal Attributes Questionnaire were hypothesized to increase vulnerability to sexual harassment. Negative Femininity, which reflects extreme passivity, and Negative Masculinity, which includes antisocial characteristics, were both found to be positively correlated with unwanted sexual experiences among male and female soldiers. PMID- 11050863 TI - Advanced classroom learning through civilian-military shareware. PMID- 11050864 TI - Challenges of including dietitians, nurses, occupational therapists, and pharmacists in the Federal Credentialing Program. AB - Credentialing and recredentialing of federal health care providers involves hundreds of hours of labor and associated costs. This article presents the history of credentialing and efforts to expand the Federal Credentialing Program to include dietitians, nurses, occupational therapists, and pharmacists and discusses barriers to this possible expansion. Representatives from federal and civilian health care service delivery agencies and credentialing and licensure bodies will gather to establish common credentialing information for these professions. Discussing barriers to these efforts will help to ensure success. In addition, a more efficient and streamlined system could easily be adopted by the civilian sector for these professions. PMID- 11050865 TI - The use of a pneumatic intermittent impulse compression device in the treatment of calcaneus fractures. AB - To determine the effects of intermittent compression on foot swelling, intracompartmental pressures, and hospital stay associated with acute calcaneus fractures, we retrospectively reviewed the records of 55 patients between January 1990 and July 1992 whose management profile included preoperative use of an intermittent compression foot pump and surgical treatment by open reduction and internal fixation. Average times were: injury to admission, 6.04 days; admission to surgery, 1.35 days; and surgery to discharge, 3.38 days. Hospital stay averaged 4.73 days. In 27 patients with suspected compartmental ischemia, admission and preoperative pressures in three compartments were averaged and compared: 18.22 and 3.81 mm Hg, respectively (p < 0.001). The authors concluded that the intermittent compression pump appears to rapidly reduce swelling of the foot and decrease elevated compartment pressures associated with calcaneus fractures, which may play a role in decreasing hospital stay. PMID- 11050866 TI - Poliomyelitis: the role of the military in the final campaign. AB - Poliomyelitis remains a disease of significance to military medicine. The medical branches of the military of many nations have much to contribute in the final 4 years of the campaign to eradicate poliomyelitis from the world. The service requirements of immunization remain a logistic charge on the defense health services of all nations. Risks to unimmunized troops remain current in the poliomyelitis endemic regions of Europe, Asia, and Africa; and recent epidemics in India, West Africa, and Albania have involved military personnel in containment programs. The 20th century has seen global attempts to eradicate seven diseases--hookworm, yellow fever, yaws, malaria, smallpox, dracunculiasis, and poliomyelitis. The first four of these were total failures, in spite of huge military logistic resources, especially in the case of yellow fever and malaria. But the global eradication of smallpox, achieved in 1979, led to the World Health Organization's Declaration of a Smallpox-Free World in 1980. Its success ranks as one of the greatest achievements in the history of medicine. Lessons learned and encouragement derived from that program led to the institution of the Poliomyelitis Global Eradication Program in 1988. Following the Declaration of a Polio-Free America, the target date for the Declaration of a Poliomyelitis-Free World has been set for 2004. Regional surveillance programs use the quality control portal of acute flaccid paralysis to monitor every potential clinical case of acute poliomyelitis. In the Western Pacific region, a region of 22 countries with a recent history of significant operational deployments, 15 countries had experienced endemic poliomyelitis before 1990. In this region, the last case of poliomyelitis (in Cambodia) was reported in March 1997. Such audit, together with massive point vaccination programs, many using massive military support, conducted since 1997 hold realistic promise that the world may be declared poliomyelitis-free by 2004. Poliomyelitis will be more difficult to eradicate than smallpox; and the current world campaign will succeed only with the logistic and professional input of the military of many nations. PMID- 11050867 TI - Uterine scar separation in patients undergoing trial of labor (TOL) in one army hospital. AB - Before the mid-1980s, repeat cesarean section was the usual method of delivering all patients who had a previous cesarean section. With cesarean section rates exceeding 20% in the 1980s, a concerted effort to reduce this high rate was formulated in all military teaching hospitals. One proposed method to decrease the overall cesarean section rate was to reduce the number of repeat cesarean sections. Trial of labor (TOL) with vaginal birth after cesarean section was one method of decreasing the high cesarean section rate. Although vaginal birth after cesarean section has a relatively high success rate and has been shown to be safe for mother and infant, TOL is not risk free. One of the potential complications of TOL is uterine scar separation, which may lead to perinatal mortality. This report discusses uterine scar separation during TOL during a 1-year period at one medium-size U.S. Army teaching hospital. PMID- 11050868 TI - Tinnitus, dizziness, and nonotologic otalgia improvement through temporomandibular disorder therapy. AB - U.S. Air Force otologic patients seeking care at Wilford Hall Medical Center for tinnitus, dizziness, and/or nonotologic otalgia without an identifiable cause and presenting with temporomandibular disorder (TMD) symptoms in the temple, jaw, or preauricular area or with otalgia at least once a month were referred to a TMD specialty clinic. The patients were provided a dental orthotic and TMD self-care instructions. After 3 months of orthotic wear, the percentages of patients reporting at least moderate symptom improvement of their tinnitus, dizziness, otalgia, and/or TMD were 64, 91, 87, and 92%, respectively. Follow-up telephone calls 6 months after completion of TMD therapy revealed that all patients maintained their symptom improvements. These findings imply that TMD was affecting the patients' otologic symptoms. Patients seeking care for tinnitus, dizziness, and/or nonotologic otalgia without an identifiable cause may have TMD, and their otologic symptoms may benefit from conservative reversible TMD therapy. PMID- 11050869 TI - Deployment dialysis in the U.S. Army: history and future challenges. AB - The U.S. Army has demonstrated that acute renal failure (ARF) could be treated successfully with dialysis during war since the early 1950s. Recent downsizing and lack of ARF patients during recent deployments may reduce the urgency to invest in the equipment modernization, personnel, and training necessary to maintain deployment dialysis capability. New dialysis equipment must be developed and purchased to replace the current Army deployable dialysis equipment that will be obsolete soon. The objective of this paper is to review ARF and dialysis during past American wars, the Armenian earthquake, and recent field training exercises to derive lessons for policy planners, clinicians, and logisticians for future deployments. Methods included medical literature search and describing the experiences of current Army personnel. The advantages and disadvantages of several commercially available dialysis systems are discussed in the context of deployment environment and policy. Recommendations for equipment and training are proposed to maintain deployment dialysis capability. PMID- 11050870 TI - Effect of universal testing machine crosshead speed on the shear bond strength and bonding failure mode of composite resin to enamel and dentin. AB - An investigation was conducted to determine if testing machine crosshead speed influenced shear bond strength (SBS) or the failure mode of composite bonded to enamel and dentin. Composite cylinders were bonded to 50 enamel and 50 dentin surfaces and thermocycled. Groups of 10 samples were debonded at speeds of 0.1, 0.5, 1.0, 5.0, and 10.0 mm/min. Data were examined with analysis of variance and post-hoc testing. Failure modes were determined using 10x magnification. With enamel, no significant differences in SBS existed, and cohesive vs. adhesive failure modes were similar for all groups. With dentin, the 0.5, 1.0, and 5.0 mm/min samples had significantly higher SBS than the 0.1 and 10.0 mm/min samples (p < 0.05). No other differences in SBS were found. Samples tested at 0.5 mm/min demonstrated strikingly better cohesive vs. adhesive results than all other groups. SBS and cohesive vs. adhesive failures achieved with dentin bonding were significantly affected by crosshead speed. PMID- 11050871 TI - Back to work more quickly after an inguinal hernia repair. AB - Tension-free hernia repair plus recovery expectancy statements return personnel to work more quickly. On the day of primary inguinal hernia repair, patients were given statements about their likelihood of returning from convalescent leave after 7 days and performing nonstrenuous work. Similar statements were given to them by telephone at 72 hours postoperatively and at a 1-week follow-up appointments. Seventy-four percent of the 73 patients returned to nonstrenuous work within 7 days, and 90% returned to strenuous work within 30 days. In this small sample, 385 work days were saved from the Navy's recommended 14 days of convalescent leave. By combining recovery expectancy statements with an effective surgical procedure, it is possible to avoid prolonged convalescence, thereby enhancing military readiness. PMID- 11050872 TI - A literature review of dental casualty rates. AB - The ability to determine dental casualty rates for the Australian Defence Force in a given situation is vital for military planners. This article reviews the literature and the available Australian Defence Force data on the subject to give some guide to planners. The review found the studies to be fairly consistent in that a well-prepared dentally fit force can expect 150 to 200 dental casualties per 1,000 soldiers per year. If the force were less prepared, as in the case of a reserve call out, this figure would be likely to increase; in the extreme case of an ill-prepared force or a force assisting in humanitarian aid, the emergency rate could be five times that figure. The literature also indicates a change in the nature of dental casualties. Although maxillofacial cases have remained steady at 25%, dental disease has decreased and endodontic cases have had a corresponding increase. PMID- 11050873 TI - Job satisfaction among nursing staff in a military health care facility. AB - Job satisfaction in the workplace affects absenteeism, turnover, and performance. We conducted a cross-sectional study of 201 nursing personnel to assess satisfaction among nursing staff at a military hospital in the southwestern United States. Participants completed a self-administered survey in which they rated professional status, autonomy, pay, organizational policies, task requirements, and interaction by importance and satisfaction. Autonomy, professional status, and pay were the most important factors and organizational policies was the least important factor. Military staff were slightly more satisfied with staff interactions than civilian staff. Nursing personnel working in specialty care units were significantly more satisfied with interactions and professional status, but they valued organizational policies less than those working in general units. Professionals were significantly more satisfied with pay and autonomy, whereas nonprofessionals were more satisfied with task requirements and professional status. PMID- 11050874 TI - Demographic, physical, and mental health factors associated with deployment of U.S. Army soldiers to the Persian Gulf. AB - A total of 675,626 active duty Army soldiers who were known to be at risk for deployment to the Persian Gulf were followed from 1980 through the Persian Gulf War. Hospitalization histories for the entire cohort and Health Risk Appraisal surveys for a subset of 374 soldiers were used to evaluate prewar distress, health, and behaviors. Deployers were less likely to have had any prewar hospitalizations or hospitalization for a condition commonly reported among Gulf War veterans or to report experiences of depression/suicidal ideation. Deployers reported greater satisfaction with life and relationships but displayed greater tendencies toward risk-taking, such as drunk driving, speeding, and failure to wear safety belts. Deployed veterans were more likely to receive hazardous duty pay and to be hospitalized for an injury than nondeployed Gulf War-era veterans. If distress is a predictor of postwar morbidity, it is likely attributable to experiences occurring during or after the war and not related to prewar exposures or health status. Postwar excess injury risk may be explained in part by a propensity for greater risk-taking, which was evident before and persisted throughout the war. PMID- 11050875 TI - Changing demographic characteristics of women veterans: results from a national sample. AB - Women veterans are a small but growing percentage of the U.S. veteran population. There are some indications that, along with this increase, the characteristics and military experiences of younger women veterans differ considerably from those of older colleagues. Many of these characteristics are not well defined, but they could have implications for women's health care needs and health policy initiatives. Using the first sample drawn from the Department of Veterans Affairs' new National Registry of Women Veterans, we designed and administered a telephone survey to a representative sample of women veterans across several major age groups. Groups approximated primary eras of military and wartime service based on the assumption that different eras might be associated with differing military experiences. We found a number of age-related similarities and differences in women veterans' demographic characteristics, military experiences, physical health symptoms, and functional outcomes. Women veterans in general also differed from female civilian counterparts on exposure to sexual trauma. Trends in the population of women veterans are likely to have implications for the variety of health care systems that treat women veterans. PMID- 11050876 TI - Cirrhosis mortality among former American prisoners of war of World War II and the Korean conflict: results of a 50-year follow-up. AB - In our earlier, 30-year follow-up of American prisoners of war (POWs) of World War II and the Korean conflict, we found evidence of increased cirrhosis mortality. Using federal records, we have now extended our follow-up to 50 years (42 years for Korean conflict veterans) and have used proportional hazards analysis to compare the mortality experience of POWs with that of controls. Compared with their controls, World War II POWs had a 32% higher risk of cirrhosis mortality (statistically significant), and mortality risk was higher in the first 30 years of follow-up and also among those aged 51 years and older. Korean POWs had roughly the same risk of cirrhosis mortality as their controls. Neither self-reported data on alcohol consumption nor supplemental morbidity data satisfactorily explained the differences in risk between POWs and controls, although there was evidence that POWs tended to have higher rates of hepatitis, helminthiasis, and nutritional deprivation. PMID- 11050877 TI - Effectiveness of a Navy remedial exercise intervention. AB - The purpose of this study was to evaluate the effectiveness of a Navy remedial exercise program (REP) in improving the physical fitness levels of its participants. Subjects were 27 Navy personnel assigned to a Northeast naval air station who had failed to pass either the body composition (circumference measures) or aerobic fitness (1.5-mile run/walk) portion of the semiannual physical readiness test. The REP consisted of three 1-hour supervised exercise sessions per week for 16 weeks, with primary emphasis on running and walking. Paired t tests revealed that the REP was effective in reducing body weight ( 3.1%) and body fat (mean absolute loss of 2.7%) and in improving 1.5-mile run/walk time (-6.2%). No changes were seen in muscular endurance measures (2 minute push-ups, 2-minute curl-ups). Findings from this study indicate that improvements in PF can be attained through a 16-week Navy remedial exercise program. PMID- 11050878 TI - The epidemiology of varicella hospitalizations in the U.S. Army. AB - Varicella infections affect the U.S. Army, but the extent has not been quantified recently. We obtained 1990 to 1997 hospitalization data from the U.S. Army Medical Command and calculated rates using data from the Army Medical Surveillance Activity and the U.S. Army Training Command. There was a decline in the number and incidence of varicella hospitalizations for U.S. Army active duty soldiers from 1990 to 1997. Varicella incidence rates for active duty soldiers are significantly higher for females, blacks, those younger than 20 years, and those whose home of record were tropical island regions. Army initial entry training hospitalizations constitute 11.8% of active duty Army hospitalizations and have also declined. Varicella continues to affect the training and health of the U.S. Army; however, the impact has diminished over the years. A feasible approach to limit varicella in the U.S. Army is to target trainees for screening or vaccination. Refinement of this strategy should be determined from a follow-up cost-effectiveness analysis. PMID- 11050880 TI - Tuberculosis of the breast presenting as carcinoma. AB - Breast infections caused by Mycobacterium tuberculosis, although rare in western countries, should not be forgotten as a cause of a breast lump presenting clinically and radiologically as a carcinoma in the older patient who gives a history of previous tuberculosis. We report the case of an 84-year-old woman with a breast lump showing noncaseating granulomas on histology who developed a sinus track after excision biopsy of the lump. The patient responded to empiric treatment with anti-tuberculosis drugs and remains well 2 years after presentation. PMID- 11050879 TI - Radiographic osseous regeneration after initial therapy with systemic doxycycline. AB - Early-onset periodontal diseases are often diagnosed in the military as a result of the requirements for annual dental examinations and the youthful population served. A young soldier diagnosed with rapidly progressive periodontitis completed initial therapy of root planing with the systemic antibiotic doxycycline but was poorly compliant with additional treatment. During a subsequent mandatory dental examination, new radiographs demonstrated a significant improvement in the quantity and quality of alveolar bone, illustrating the regeneration potential of the young patient with early-onset periodontal disease. PMID- 11050881 TI - [Stomach evil Helicobacter pylori. A misunderstood benefactor?]. PMID- 11050882 TI - [Eradication euphoria is passe. Does H. pylori protect against reflux?. Interview by Dr. Brigitte Moreano]. PMID- 11050883 TI - [New preventive antimalaria drug. 3 tables, 2 months of protection. Interview by Petra Eiden]. PMID- 11050885 TI - [Every second patient with chronic pain is also depressive. Tricyclic antidepressants and cognitive behavior therapy are indicated]. PMID- 11050884 TI - [Paradoxical therapy in tinnitus. Acoustic stimulator improves tinnitus]. PMID- 11050886 TI - [When the mind affects the body. Unmasking "masked" depression]. AB - The term masked depression means a depressive condition in which physical symptoms predominate while the depressive symptoms are concealed. The physical complaints are frequently unspecific and difficult to identify. The leading symptom is usually pain--the predilection sites being the head and chest. For the general practitioner, orientation to a check list of questions aimed at identifying depressive symptoms in patients with physical complaints that are difficult to diagnose, makes good sense. The concept of masked depression has been abandoned in favor of other diagnoses that can be operationalized. PMID- 11050888 TI - [Insulin sensitizer. A new therapy option for type 2 diabetic patients]. AB - The glitazones, also known as insulin sensitizers, represent a new concept in the treatment of type 2 diabetes. While previously available antidiabetics have no influence on an existing insulin resistance, the glitazones act directly on the receptors in the muscles, fatty tissue and liver, and in this way reduce insulin resistance. To date, three members of this group of substances are known: troglitazone, rosiglitazone and pioglitazone. In a study done to investigate pioglitazone, fasting blood sugar and HbA1c achieved virtually normal values, and lipid metabolism also improved. On the basis of the results of studies done to date, the tolerability of both pioglitazone and rosiglitazone is good. In contrast to troglitazone, no evidence has been found for a hepatotoxic effect. Insulin sensitizers do not cause hypoglycemia, since they do not stimulate the pancreas. They are suitable both for monotherapy and combination treatment with metformin, sulfonylureas and insulin. PMID- 11050887 TI - [Treatment of chronic and neuropathic pain. Established amitriptyline and the new gabapentin]. PMID- 11050890 TI - [Arthroses. 2: Various forms of therapy]. PMID- 11050889 TI - [Physician-patient interaction in hypertension. Partnership with defined roles]. PMID- 11050891 TI - [Exanthema with dyspnea. Varicella with interstitial pneumonia]. PMID- 11050892 TI - [11. Venous thrombosis--risk factors and therapy]. PMID- 11050893 TI - [The operating microscope in dental general practice]. AB - The great advantages of using a stereoscopic operating microscope in endodontics and especially in endodontic surgery are being increasingly advocated in postgraduate training courses and numerous publications. Widely underestimated, however, is the fact that this tool may open new horizons to almost all dental specialities and will improve ergonomics of the dental work place. Moreover, the superb images may contribute some pleasure to the daily dental work and stress. All this makes the clinical stereoscope interesting also for the general practice of dentistry. The purpose of this paper is to report about several years of practical experience with the "scope" in general dentistry. Technical aspects are discussed as well as the possibilities of use. Practical hints regarding the type of scope and the technique of its use are given in order to lower the barrier of entrance into this new era of doing dental work. PMID- 11050894 TI - [Amalgam and other restoration materials. The opinion of dentists in the 3 linguistic regions of Switzerland (2)]. PMID- 11050895 TI - Progress towards poliomyelitis eradication, Ethiopia. PMID- 11050896 TI - Malaria--economic implications. PMID- 11050897 TI - Rift Valley fever, Saudi Arabia (update). PMID- 11050898 TI - Yellow fever, 1998-1999. PMID- 11050899 TI - Getting health back on track. PMID- 11050900 TI - Services and information utilised by female sex workers for sexual and physical safety. AB - AIMS: To examine services utilised by female sex workers in Christchurch for sexual and physical safety. METHOD: Estimates were made of the sex worker population in Christchurch before conducting a cross-sectional survey. RESULTS: 303 women responded. Almost all went for sexual health check-ups. Most (251) had a general practitioner, but only about half disclosed they were sex workers. Of the 135 women who used their own general practitioner for sexual health checks, 62% disclosed they were sex workers. In general, the women relied upon informal networks for information and advice, but the peer organisation of New Zealand Prostitutes Collective was also important. CONCLUSION: While general practitioners were used by the majority of sex workers, high levels of nondisclosure need to be understood and redressed. The preference of sex workers for informal, peer relationships should be considered in future health promotion efforts. PMID- 11050901 TI - The early experience of general practitioners using Green Prescription. AB - AIM: Sedentary lifestyle is a significant risk factor for increased morbidity and mortality in many medical conditions. A Hillary Commission initiative, Green Prescription is a written exercise prescription given by general practitioners (GPs) to sedentary patients to encourage physical activity. Our aim was to establish the extent to which GPs in the North Health region in 1997 issued with Green Prescription packages had used them, the circumstances under which they were used, and barriers to their use. METHODS: 433 GPs issued with packs were faxed a one-page questionnaire for immediate completion, with follow-up of non responders. RESULTS: The response rate was 73%, with 65% of respondents having written Green Prescriptions. Their main reasons for use were patient need for more exercise and presence of high-risk medical conditions such as hypertension, cardiovascular disease, obesity and diabetes. Reasons for non-use were: GP already giving advice about physical activity; concern that Green Prescription was patronising and simplistic; compliance issues and time restraints. Some requested a computerised version. CONCLUSION: Non-responders may be non-users, hence we estimate that 48-65% of targeted GPs used Green Prescription. Barriers identified by GPs have assisted in Green Prescription development, which is now nationwide and assessed by independent researchers tri-annually. PMID- 11050902 TI - Characteristics of children with florid vitamin D deficient rickets in the Auckland region in 1998. AB - AIM: To describe the characteristics of children with vitamin D deficiency rickets and identify common features and predisposing factors. METHODS: A review of the clinical notes of all children less than five years of age with radiological evidence of rickets and serum 25-hydroxyvitamin D levels of less than 10 micrograms/L. Patients were identified by searching all low vitamin D levels performed at the Endocrinology laboratory at Auckland Hospital and children presenting to the Starship Childrens' Hospital with rickets in 1998. RESULTS: In 1998, there were eighteen children (ten males and eight females) with vitamin D deficient rickets. The age range was 3 to 36 months with a median of 12 months. There were twelve children of Indian ethnic origin, one Maori, one Tongan, one Western Samoan, one Ethiopian, one Moroccan and one Indonesian. All children had an elevated alkaline phosphatase level and most had very low serum 25-hydroxyvitamin D levels (< or = 5 micrograms/L), and over half were hypocalcaemic. The common presenting features were delayed walking and bowed legs, swollen wrists or ankles, hypocalcaemic seizure, incidental radiological abnormalities and failure to thrive. CONCLUSIONS: There are a significant number of children in Auckland presenting with florid clinical rickets. The majority with vitamin D deficient rickets in this survey were of Indian ethnic origin. Strategies are needed to detect children at risk of vitamin D deficiency and supplement them with vitamin D. PMID- 11050903 TI - Extracorporeal shock wave lithotripsy for difficult common bile duct stones: initial New Zealand experience. AB - AIM: Common bile duct (CBD) stones can usually be managed by open surgery, endoscopic retrograde cholangiopancreatography (ERCP) or radiological intervention. At times, however, these methods are either unsuccessful or inappropriate. We report our initial experience of extracorporeal shock wave lithotripsy (ESWL) for CBD stones that had either been unsuccessfully managed by conventional techniques, or in cases where these techniques were associated with a high level of risk. METHODS: A retrospective review of medical records of cases receiving ESWL for CBD was undertaken. The aspects reviewed were: indications, outcome and completions from the procedure. RESULTS: ESWL was used in the management of eight patients (three male, five female, age range 24-83, mean 54 years). The indications in five cases were failure of open surgery, ERCP or radiological techniques to clear the duct. In the other three cases, ERCP was unsuccessful and there was significant coincidental medical illness (morbid obesity with diabetes, and severe ischaemic heart disease). CBD clearance was achieved in seven cases. In one unsuccessful case, the duct was cleared after two open procedures. CONCLUSIONS: ESWL can be used to clear CBD stones. It should only be used, however, where prior CBD drainage has been achieved, preferably by endoscopic sphincterotomy. Morbid obesity is a relative contraindication to the use of ESWL. If ESWL fails, a period of time should be allowed to elapse before open surgery because of distortion of soft tissue planes. ESWL can be a useful technique in dealing with some difficult CBD stones. PMID- 11050904 TI - Follow up testing of hyperglycaemia during hospital admission: combined use of fasting plasma glucose and HbA1c. AB - AIM: To follow up patients without known diabetes, but with hyperglycaemia in hospital for diabetes at one year. METHODS: 159 patients with a random plasma glucose > or = 7.8 mmol/L recorded during hospital admission were sent a questionnaire and invited to have the following test one year following discharge: fasting plasma glucose, HbA1c and fasting lipid profile. Those with a fasting plasma glucose > or = 5.5 and < 7.0 mmol/L, and/or those with a HbA1c > or = 6.0%, were asked to have an oral glucose tolerance test. Those with a fasting plasma glucose > or = 7.0 mmol/L were defined as having diabetes. RESULTS: There were 88 full responses. Nineteen (21.6%) had diabetes and nine impaired glucose tolerance. Hb1Ac was > or = 6% in five subjects with a fasting plasma glucose < 5.5 mmol/L. Two had impaired glucose tolerance and one diabetes. If a random plasma glucose in-hospital of 10 mmol/L is used as a threshold for later testing, as suggested by previous studies, then 25% of those with an abnormal result would have been missed. CONCLUSIONS: A high proportion of those with hyperglycaemia in hospital have diabetes or impaired glucose tolerance at one year. Initial testing with fasting plasma glucose and HbA1c avoided oral glucose tolerance test in 76% of cases. Use of HbA1c detected otherwise missed diabetes and impaired glucose tolerance. A random plasma glucose of > or = 7.8 mmol/L in hospital targets patients who should be tested for impaired glucose tolerance or diabetes following discharge. PMID- 11050905 TI - Coronary angioplasty in New Zealand 1995-1998: a report from the National Coronary Angioplasty Registry. AB - AIMS: To report coronary angioplasty data collected by the New Zealand Coronary Angioplasty Registry from 1995-1998. METHOD: Information on all patients undergoing attempted coronary angioplasty in eight New Zealand institutions was recorded on datasheets at the time of, or soon after, the procedure. These were forwarded to the registry at Green Lane Hospital. RESULTS: Over the four-year period, 8395 angioplasty procedures were performed by 26 cardiologists in eight coronary interventional facilities, with a procedural success rate of 94%. Procedural numbers grew steadily, with 55% more coronary angioplasties performed in 1998 than in 1995 (p = 0.02). The New Zealand national angioplasty rate, which rose from 459/million population in 1995 to 684/million in 1998, remains lower than that of Australia and Western European countries. Excluding those that underwent angioplasty for acute myocardial infarction, the number of peri procedural deaths was similar, with six in 1995 and four in 1998 (p = 0.30), and the requirement for emergency bypass surgery fell from 22 cases in 1995 to three in 1998 (p < 0.001). The use of stents increased dramatically, with 85% of patients receiving a stent in 1998, compared with 23% in 1995 (370% increase, p < 0.001). This was associated with a reduction in the number of patients requiring repeat percutaneous interventions for restenosis (10.7% in 1995 to 6.4% in 1998, p < 0.001). CONCLUSION: There has been a steady growth in the numbers of patients with coronary artery disease treated by coronary angioplasty, and in the number treated by intracoronary stents from 1995 to 1998. The need for urgent coronary bypass surgery has fallen. Continued submission of complete and accurate data to the coronary angioplasty registry is vital for ongoing audit. PMID- 11050906 TI - Re-using 'single-use' medical devices. PMID- 11050907 TI - Oral contraceptives and venous thromboembolism. PMID- 11050908 TI - Safety of doctors undertaking domiciliary visits--policy of Occupation Safety and Health Service (OSH) PMID- 11050909 TI - Accuracy of death certification. PMID- 11050910 TI - Professional misconduct. PMID- 11050911 TI - Medical Council of New Zealand. Guidelines on Responsibilities of Doctors in Management and Governance. PMID- 11050913 TI - Radiotherapy for hepatocellular carcinoma. AB - PURPOSE: To retrospectively evaluate the effectiveness of radiotherapy with or without transarterial embolization (TAE) and/or percutaneous ethanol injection (PEI) in patients with hepatocellular carcinoma who were ineligible for surgery. PATIENTS AND METHODS: From October 1984 to November 1997, 62 patients underwent radiotherapy receiving 50 to 70 Gy in 25 to 35 treatments with or without transarterial embolization and/or percutaneous ethanol injection and were followed for a median period of 8.6 months (1.5 to 92 months). RESULTS: Overall median survival rates were 9.5 months. Significant prognostic factors were the extent of pretreatment liver function impairment, radiation field size and the existence of tumor thrombosis. Six-month and 1-year local control rates were 67 and 54%, respectively. Seven of the 8 patients who suffered from hepatic failure had poor pretreatment liver functions. CONCLUSION: Radiotherapy with or without transarterial embolization and/or percutaneous ethanol injection appears effective in controlling hepatocellular carcinoma and prolonged survival. Individualized treatment strategies are presented depending on the tumor presentation and the degree of liver function impairment. PMID- 11050912 TI - [Interdisciplinary consensus on diagnosis and therapy of testicular tumors. Results of an update conference based on evidence-based medicine. German Testicular Cancer study Group (GTCSG)]. AB - BACKGROUND: An "Interdisciplinary Consensus Statement on the Diagnosis and Therapy of Testicular Tumors" was prepared in 1996 by the "Interdisciplinary Testicular Tumor Working Group" (IAH) with input from representatives from diagnostic and therapeutic disciplines of various working groups of the German Cancer Society (Strahlenther Onkol 1997;173:397-406). In 1998 the IAH met again together with the "Testicular Tumor Working Party" of the Urooncology Working Group (AUO) and formed the "German Testicular Cancer Study Group" (GTCSG). Defined and accepted interdisciplinary standards from the initial meeting were revised based on current scientific developments and clinical results. This cooperating effort increased the quality of the initial recommendations and helped to put the recommendations for diagnosing and treating testicular tumor on a broader scientific basis. METHODS: According to the principles of "evidence based medicine" (EBM), the Consensus from 1996 was modified, based on the current level of evidence from the published literature. The methodological process and evaluation criteria used were that of the "Cochrane Collaboration". RESULTS: An "Interdisciplinary Update Consensus Statement" summarizes and defines the diagnostic and therapeutic standards according to the current scientific practices in testicular cancer. For 21 separate areas scientifically based decision criteria are suggested. For treatment areas where more than one option exist without a consensus being reached for a preferred strategy, such as in seminoma in clinical Stage I or in non-seminoma Stages CS I or CS IIA/B, all acceptable alternative strategies with their respective advantages and disadvantages are presented. This "Interdisciplinary Update Consensus" was presented at the 24th National Congress of the German Cancer Society on March 21st and subsequently evaluated and approved by the various German scientific medical societies. PMID- 11050914 TI - Remission rates following preoperative chemotherapy and radiation therapy in patients with breast cancer. AB - PURPOSE: To evaluate remission and breast-conservation rates after preoperative chemotherapy or chemo-radiotherapy (CT-RT). PATIENTS AND METHODS: Seventy-three patients with 74 biopsy-proven invasive breast cancers prospectively entered the protocol. Eighteen patients were treated by neoadjuvant chemotherapy followed by surgery and adjuvant irradiation (chemotherapy group). Fifty-five patients with 56 tumors were treated with combined neoadjuvant chemo-radiotherapy, followed by surgery (chemo-radiotherapy group). Most patients of both treatment groups received 4 cycles of EC chemotherapy. In some patients with large tumors 3 cycles of CMF were added. Chemotherapy was followed by hormonal treatment with tamoxifen or LHRH agonists in case of positive hormone-receptor status. Preoperative radiotherapy was administered using 2 Gy fractions up to a total dose of 50 Gy, followed by a tumor boost of 6 to 11 Gy. The median overall treatment time was 41 days (range: 35 to 55 days). The median time interval between end of neoadjuvant therapy and surgery was 11 weeks (range: 10 to 22 weeks) and 27 weeks (range: 11 to 41 weeks) for the chemotherapy- and chemo-radiotherapy group. The median time interval between end of chemotherapy and the beginning of irradiation ranged between 2 and 8 weeks (median 4 weeks) in the chemo-radiotherapy group. RESULTS: Side-effects due to chemo- or radiotherapy were moderate and reversible. In the chemotherapy group 17/18 patients (94%) achieved a partial (pPR) and 1/18 patients (6%) a complete histopathological response (pCR). In the chemo radiotherapy group 32/56 (57%) showed a pPR and 24/56 (43%) a pCR. The difference in complete remission is significant (Fisher's Exact Test: p = 0.004). In 45/74 cases (61%) the breast was preserved, immediate breast reconstructions with rectus myocutaneous flaps (TRAM) after mastectomy were performed in 8/74 cases (11%) and modified radical mastectomies without reconstruction were required in 21/74 cases (28%). The breast conservation rates were similar in both treatment groups. CONCLUSIONS: Even though the small number of patients in the present protocol does not permit definite conclusions, the results of combined modality treatment seem promising with regard to tumor remission within the treated breast and as a tool for breast conservation in advanced stage disease. On the basis of these encouraging data a prospective Phase-III study has been initiated. PMID- 11050915 TI - Intermittent use of amifostine during postoperative radiochemotherapy and acute toxicity in rectal cancer patients. AB - PURPOSE: Amifostine has been shown to be able to reduce acute radiation toxicity if administered daily prior to radiation during a course of a conventionally fractionated radiotherapy. A disadvantage is the necessity of daily intravenous injection. We have used amifostin in patients undergoing adjuvant radiochemotherapy for rectal cancer. Amifostine was administered only in the first and fifth week of radiotherapy together with 5-FU chemotherapy. The objective was to determine whether the intermittent use of amifostine may be effective in reducing acute radiation toxicity. PATIENTS AND METHODS: From September 1997 through October 1998, 30 patients with stage II/III rectal cancer underwent postoperative radiochemotherapy at our department. All patients had undergone curative (R0) resection and received 50.4 Gy to the pelvis with a 3 field technique using a belly board followed by a boost of 5.4 Gy to the presacral space in conventional fractionation with 1.8 Gy per fraction. 5-FU chemotherapy was administered as 120-hours continuous infusion in the first and fifth radiation week via a central venous catheter in a daily dosage of 1,000 mg/m2. All patients were offered to participate in a phase-II study using additional amifostine. Fifteen patients participated and received 500 mg amifostine daily on chemotherapy days (days 1 to 5 and 29 to 33) immediately prior to the daily radiation fraction. Fifteen patients did not participate and served as non-randomized control. The study was approved by the ethical committee of the Martin-Luther-University and informed consent was obtained from all patients. RESULTS: The distribution of patients' characteristics and prognostic parameters was comparable in both groups. Side effects of amifostine were mild and included hypotension (53% grade I, 7% grade II) and nausea (47% grade I, 13% grade II). Antiemetics were not routinely used. All patients completed radiochemotherapy plus amifostine without unplanned breaks or dose reductions. One patient developed a cerebral infarction which was considered to be not related to the use of amifostine. As compared to the non-randomized control group, patients with additional amifostine had less acute skin and bowel toxicity (maximum erythema score 1.47 +/- 0.64 without vs 0.87 +/- 0.52 with amifostine, p = 0.009 and maximum diarrhea score 1.07 +/- 1.03 vs 0.40 +/- 0.63, p = 0.044). Oral 5-FU-related mucositis and hematological toxicity were not significantly different. CONCLUSIONS: In this phase-II study, amifostine significantly reduced acute skin and bowel toxicity of adjuvant chemoradiation in patients with rectal cancer even if the drug was administered only intermittently and not during the whole course of radiotherapy. This finding might be important with regard to intense combined regimes and should be further investigated. PMID- 11050916 TI - Effect of doxorubicin on cell survival and micronuclei formation in HeLa cells exposed to different doses of gamma-radiation. AB - PURPOSE: The present study was undertaken to obtain an insight into the combined effects of doxorubicin with radiation on the cell survival and micronuclei induction in HeLa cells. MATERIAL AND METHODS: HeLa S3 cells were allowed to grow till they reached plateau phase, inoculated with 10 micrograms/ml doxorubicin hydrochloride and then exposed to 0, 0.5, 1, 2 and 3 Gy gamma-radiation. Clonogenicity of cells was measured using the colony forming assay, micronuclei formation using the micronucleus assay. RESULTS: The treatment of HeLa cells with doxorubicin (adriamycin) for 2 hours before exposure to different doses of gamma radiation resulted in a significant and dose-dependent decline in the cell survival and cell proliferation when compared to the PBS + irradiation group. Conversely, the frequency of micronuclei increased in a dose-related manner in both the PBS + irradiation and doxorubicin + irradiation groups. The pretreatment of HeLa cells with doxorubicin before irradiation to various doses of gamma-rays resulted in a significant elevation in the frequency of micronuclei when compared with the concurrent PBS + irradiation group. The dose-response relationship for both PBS + irradiation and doxorubicin + irradiation groups was linear. The correlation between cell survival and micronuclei induction was also determined for PBS or doxorubicin + irradiation group, where the clonogenicity of cells declined with the increase in micronuclei formation. The correlation between cell survival and micronuclei induction was linear quadratic for both PBS + irradiation and doxorubicin + irradiation groups. CONCLUSION: From our study it can be concluded that combination treatment with doxorubicin and radiation increased the genotoxic effect of the either treatment given alone. PMID- 11050917 TI - Daily CT planning during boost irradiation of prostate cancer. Feasibility and time requirements. AB - BACKGROUND: In the irradiation of prostate cancer internal organ movement leads to uncertainties in the daily localization of the clinical target volume. Therefore more or less large safety margins are added when designing the treatment portals. With daily CT planning internal organ movement can be compensated to some extent, safety margins can be reduced and irradiated normal tissue can be spared. The feasibility of daily CT-based 3D treatment planning is studied in a patient with localized prostate carcinoma using a new patient positioning system. METHODS: Daily CT planning was applied during boost irradiation of a patient with prostate cancer: After patient immobilization the pelvis was scanned in 3 mm CT slices. Planning was done with the BrainSCAN planning system for stereotactic body irradiation. The prostate was contoured in all slices and the safety margins of the micromultileafs were automatically set to the distance chosen by the physician (0.8 cm). Patient positioning was done with the BrainLAB ExacTrac positioning system on the basis of skin attached stereotactic body markers. Before each treatment verification images of the isocenter were taken. RESULTS: The total time requirement for planning and irradiation was about 1 hour 15 minutes. Patient positioning on the treatment couch took about 10 minutes. The accuracy of the positioning system was good (75% of the deviations were smaller than 3 mm). The shift of the single markers from CT scan to CT scan was more extensive than those of the center of all 7 markers combined (47% of the deviations were smaller than 3 mm). The location of the markers seems to influence the magnitude of their dislocation. CONCLUSION: Daily CT planning is feasible but time consuming. The new patient positioning system ExacTrac is an interesting tool especially for daily CT planning since conventional simulation can be omitted. PMID- 11050918 TI - Are local recurrences after breast conserving therapy more frequent in patients with BRCA1/BRCA2 mutations? PMID- 11050919 TI - [Meta-analysis confirms improved survival rate by locoregional radiotherapy of breast carcinoma]. PMID- 11050920 TI - [Paclitaxel versus doxorubicin in first-line therapy of metastatic breast carcinoma]. PMID- 11050921 TI - [Effects of radiotherapy and chemotherapy on lung function in patients with breast neoplasm and lymphoma: a follow-up study]. PMID- 11050922 TI - EUROTOX 2000. 17-20 September 2000. London. Abstracts. PMID- 11050924 TI - 16th International Congress on Thrombosis. Porto, Portugal, May 5-8, 2000. Abstracts. PMID- 11050923 TI - International Society for Digestive Surgery (ISDS) 17th World Congress. September 6-9, 2000. Hamburg, Germany. Abstracts. PMID- 11050925 TI - The American Society for Histocompatibility and Immunogenetics 26th annual meeting. October 10-14, 2000. Lake Buena Vista, Florida, USA. Abstracts. PMID- 11050926 TI - 14th European Immunology Meeting. EFIS 2000. 23-27 September 2000. Abstracts. PMID- 11050927 TI - European Virology 2000. 17-21 September 2000, Glasgow, Scotland. Abstracts. PMID- 11050928 TI - 15th Congress of the European Sleep Research Society. 12-16 September 2000. Istanbul, Turkey. Abstracts. PMID- 11050929 TI - 2nd International Congress on Hormones, Brain and Neuropsychopharmacology. Rhodes, Greece. July 15-19, 2000. Abstracts. PMID- 11050930 TI - Advances in protein chemistry. Introduction. PMID- 11050931 TI - New functions from old scaffolds: how nature reengineers enzymes for new functions. PMID- 11050932 TI - Evolution of protein function by domain swapping. PMID- 11050933 TI - Rational evolutionary design: the theory of in vitro protein evolution. AB - Directed evolution uses a combination of powerful search techniques to generate proteins with improved properties. Part of the success is due to the stochastic element of random mutagenesis; improvements can be made without a detailed description of the complex interactions that constitute function or stability. However, optimization is not a conglomeration of random processes. Rather, it requires both knowledge of the system that is being optimized and a logical series of techniques that best explores the pathways of evolution (Eigen et al., 1988). The weighing of parameters associated with mutation, recombination, and screening to achieve the maximum fitness improvement is the beginning of rational evolutionary design. The optimal mutation rate is strongly influenced by the finite number of mutants that can be screened. A smooth fitness landscape implies that many mutations can be accumulated without disrupting the fitness. This has the effect of lowering the required library size to sample a higher mutation rate. As the sequence ascends the fitness landscape, the optimal mutation rate decreases as the probability of discovering improved mutations also decreases. Highly coupled regions require that many mutations be simultaneously made to generate a positive mutant. Therefore, positive mutations are discovered at uncoupled positions as the fitness of the parent increases. The benefit of recombination is twofold: it combines good mutations and searches more sequence space in a meaningful way. Recombination is most beneficial when the number of mutants that can be screened is limited and the landscape is of an intermediate ruggedness. The structure of schema in proteins leads to the conclusion that many cut points are required. The number of parents and their sequence identity are determined by the balance between exploration and exploitation. Many disparate parents can explore more space, but at the risk of losing information. The required screening effort is related to the number of uphill paths, which decreases more rapidly for rugged landscapes. Noise in the fitness measurements causes a dramatic increase in the required mutant library size, thus implying a smaller optimal mutation rate. Because of strict limitations on the number of mutants that can be screened, there is motivation to optimize the content of the mutant library. By restricting mutations to regions of the gene that are expected to show improvement, a greater return can be made with the same number of mutants. Initial studies with subtilisin E have shown that structurally tolerant positions tend to be where positive activity mutants are made during directed evolution. Mutant fitness information is produced by the screening step that has the potential to provide insight into the structure of the fitness landscape, thus aiding the setting of experimental parameters. By analyzing the mutant fitness distribution and targeting specific regions of the sequence, in vitro evolution can be accelerated. However, when expediting the search, there is a trade-off between rapid improvement and the quality of the long-term solution. The benefit of neutrality has yet to be captured with in vitro protein evolution. Neutral theory predicts the punctuated emergence of novel structure and function, however, with current methods, the required time scale is not feasible. Utilizing neutral evolution to accelerate the discovery of new functional and structural solutions requires a theory that predicts the behavior of mutational pathways between networks. Because the transition from neutral to adaptive evolution requires a multi-mutational switch, increasing the mutation rate decreases the time required for a punctuated change to occur. By limiting the search to the less coupled region of the sequence (smooth portion of the fitness landscape), the required larger mutation rate can be tolerated. Advances in directed evolution will be achieved when the driving forces behind such proce PMID- 11050934 TI - Temperature adaptation of enzymes: lessons from laboratory evolution. PMID- 11050935 TI - Structural analysis of affinity matured antibodies and laboratory-evolved enzymes. PMID- 11050937 TI - Analysis of large libraries of protein mutants using flow cytometry. PMID- 11050936 TI - Molecular breeding: the natural approach to protein design. PMID- 11050938 TI - From catalytic asymmetric synthesis to the transcriptional regulation of genes: in vivo and in vitro evolution of proteins. PMID- 11050939 TI - In vitro selection and evolution of proteins. PMID- 11050940 TI - DNA vaccines for viral infections: basic studies and applications. PMID- 11050941 TI - SV40 large T antigen functions in DNA replication and transformation. PMID- 11050942 TI - Viral and cellular mRNA capping: past and prospects. PMID- 11050943 TI - Mechanism of genome transcription in segmented dsRNA viruses. AB - Genome transcription is a critical stage in the life cycle of a virus, as this is the process by which the viral genetic information is presented to the host cell protein synthesis machinery for the production of the viral proteins needed for genome replication and progeny virion assembly. Viruses with dsRNA genomes face a particular challenge in that host cells do not produce proteins which can transcribe from a dsRNA template. Therefore, dsRNA viruses contain all of the necessary enzymatic machinery to synthesize complete mRNA transcripts within the core without the need for disassembly. Indeed one of the more striking observations about genome transcription in dsRNA viruses is that this process occurs efficiently only when the transcriptionally competent particle is fully intact. This observation suggests that all of the components of the TCP, including the viral genome, the transcription enzymes, and the viral capsid, function together to produce and release mRNA transcripts and that each component has a specific and critical role to play in promoting the efficiency of this process. This review has examined the process of genome transcription in dsRNA viruses from the perspective of rotavirus as a model system. However, despite numerous architectural and organizational differences among the families of dsRNA viruses, numerous studies suggest that the basic mechanism of mRNA production may be similar in most, if not all, viruses having dsRNA genomes. Important functional similarities include (1) the presence of a capsid-bound RNA-dependent RNA polymerase, which produces single-stranded mRNA transcripts from the dsRNA genome and regenerates the dsRNA genome from single-stranded RNA templates; (2) in viruses infecting eukaryotic hosts, the presence of all the enzymatic activities needed to generate the 5' cap required by the eukaryotic translation machinery; (3) the high degree of structural order present in the packaged genome, suggesting the requirement for organization in the viral core; (4) the role of the innermost capsid protein as a scaffold on which the core components of the transcription apparatus are assembled; and (5) the release of nascent mRNA transcripts through channels at the icosahedral vertices. The process of genome transcription in dsRNA viruses will become better understood as structural studies progress to higher resolution and as more viruses become amenable to study using site-directed mutagenesis coupled with viral reconstitution to generate recombinant particles having precise functional and structural changes. Future studies will dissect important intermolecular interactions required for efficient mRNA synthesis and will shed further light on the reasons for which the viral core must be structurally intact in order for transcription to occur efficiently. Structural studies of the capping enzymes at atomic resolution will reveal how multiple enzyme activities reside within a single polypeptide and how they act in concert to synthesize the 5' cap on the end of each mature transcript. Perhaps most interestingly, high resolution structural studies of actively transcribing virions will provide insight into the conformational changes that occur within the core during mRNA synthesis. Together, these studies will clarify the function of this complex macromolecular machine and will also shed additional light on the basic principles of virus architecture and assembly, as well as provide avenues for the design of antiviral therapies. PMID- 11050944 TI - Structures and mechanisms in flavivirus fusion. PMID- 11050945 TI - Avsunviroidae family: viroids containing hammerhead ribozymes. PMID- 11050946 TI - Virus resistance mediated by ribosome inactivating proteins. PMID- 11050948 TI - Perspectives on viral vector design and applications. PMID- 11050947 TI - Groundnut rosette disease virus complex: biology and molecular biology. PMID- 11050949 TI - Replication-competent herpes simplex viral vectors for cancer therapy. PMID- 11050950 TI - Herpes simplex virus type 1-based amplicon vector systems. PMID- 11050951 TI - Epstein-Barr virus vectors for gene therapy. PMID- 11050952 TI - Cytomegalovirus bacterial artificial chromosomes: a new herpesvirus vector approach. PMID- 11050953 TI - Adenovirus vectors for human gene therapy. PMID- 11050955 TI - Production of recombinant adeno-associated virus. AB - Currently, rAAV appears to be one of the most promising vectors for gene therapy applications. Attractive features of the vector include nonpathogenicity, the ability to infect nondividing cells, escape from host immune responses, and integration into the host genome. Tremendous progress has been made in the production of this vector, which makes it possible to start to examine the vector performance in large animals and to implement the transition to phase I human clinical trials with a variety of target tissues and therapeutic genes. However, some major challenges remain to be addressed by more extensive studies. These include the current inability to provide rAAV vectors in sufficient quantity and purity for large-scale clinical human applications, lack of site-specific integration, and lack of efficient transduction in some tissues such as airway epithelial cells. There is a limited transgene capacity in recombinant virus particles, and repeated administration of the vectors may be necessary to treat patients with chronic forms of genetic disease. Nevertheless, it is reasonable to assume that significant refinements will be made in all these areas in the relatively near future. This will promote the potential for successful therapeutic applications in humans, using rAAV-mediated gene transfer for a variety of different diseases. PMID- 11050954 TI - Adeno-associated virus vector-mediated gene transfer to somatic cells in the central nervous system. PMID- 11050956 TI - Retroviral vectors. AB - Retroviral vectors are widely used for preclinical and clinical applications. Unlike many of the other types of vectors currently being developed for gene therapy, retroviral vectors are able to genetically modify cells stably without perturbing cell growth. Retroviral vectors based on murine retroviruses are well suited for ex vivo applications where the cells are rapidly dividing. In particular, retroviral viral vectors have been used for a variety of ex vivo gene therapy approaches for treating genetic diseases such as Gaucher and severe combined immunodeficiency (SCID) and for acquired diseases such as cancer and arthritis. However, recent advances in the production of retroviral vectors have allowed for their use in vivo such as for the treatment of cancer and human immunodeficiency virus. The ability to target retroviral vectors to specific cell types will also increase the utility of high titer retroviral vectors for in vivo applications. Clearly retroviral vectors have been extremely useful for both preclinical and clinical gene therapy studies, and it is likely that they will continue to be utilized for ex vivo and in vivo strategies in the future. PMID- 11050957 TI - Alphavirus expression vectors. PMID- 11050958 TI - Transfectant influenza viruses as antigen delivery vectors. PMID- 11050959 TI - Lentiviral vectors. PMID- 11050960 TI - [How well-positioned is a publication in the German language?]. PMID- 11050961 TI - [Anesthesia in bronchial asthma]. AB - Asthma is defined as a chronic inflammatory airway disease in response to a wide variety of provoking stimuli. Characteristic clinical symptoms of asthma are bronchial hyperreactivity, reversible airway obstruction, wheezing and dyspnea. Asthma presents a major public health problem with increasing prevalence rates and severity worldwide. Despite major advances in our understanding of the clinical management of asthmatic patients, it remains a challenging population for anesthesiologists in clinical practice. The anesthesiologist's responsibility starts with the preoperative assessment and evaluation of the pulmonary function. For patients with asthma who currently have no symptoms, the risk of perioperative respiratory complications is extremely low. Therefore, pulmonary function should be optimized preoperatively and airway obstruction should be controlled by using steroids and bronchodilators. Preoperative spirometry is a simple means of assessing presence and severity of airway obstruction as well as the degree of reversibility in response to bronchodilator therapy. An increase of 15% in FEV1 is considered clinically significant. Most asymptomatic persons with asthma can safely undergo general anesthesia with and without endotracheal intubation. Volatile anesthetics are still recommended for general anesthetic techniques. As compared to barbiturates and even ketamine, propofol is considered to be the agent of choice for induction of anesthesia in asthmatics. The use of regional anesthesia does not reduce perioperative respiratory complications in asymptomatic asthmatics, whereas it is advantageous in symptomatic patients. Pregnant asthmatic and parturients undergoing anesthesia are at increased risk, especially if regional anesthetic techniques are not suitable and prostaglandin and its derivates are administered for abortion or operative delivery. Bronchial hyperreactivity associated with asthma is an important risk factor of perioperative bronchospasm. The occurrence of this potentially life-threatening condition in anesthesia practice varies from 0.17 to 4.2%. The anesthesiologists' goal should be to minimize the risk of inciting bronchospasm and to avoid triggering stimuli. As increases in airway resistance are noticed, therapy should be directed towards optimizing oxygenation and proper diagnosis needs to be established. With deepening anesthesia level and aggressive pharmacological management utilizing both, beta-agonists and steroids, respiratory failure may be properly controlled. PMID- 11050962 TI - [German language publications of German university departments of anesthesiology]. AB - OBJECTIVE: To evaluate contributions of German university departments of anesthesiology to the German language literature. METHODS: Using Medline (Silverplatter, volumes 1988-1997 of the 1999 edition) we searched for (journal article IN SO) AND ((anesth* OR anasth* OR anaesth*) in AD) AND (LA-German). The publications found were manually evaluated for address. The "Jahrbuch der Anasthesiologie und Intensivmedizin" (1992-1995) was evaluated for the number of staff members (anaesthesiologists, residents and non-physician scientists). RESULTS: The total number of publications was about 200/year. Individual departments contributed very differently (range: 0.6 publications/year to 14.8 publications/year). The department with the highest per capita output had about 0.4 Medline-indexed papers per scientist and year, the department with the lowest per capita output had about 0.02 papers per scientist and year. CONCLUSION: There is a great difference with respect to total publication count. The differences in total publication count can only in part be explained by different size of the departments. Even the leading department had far less than 1 paper per scientist and year. PMID- 11050963 TI - Performance of the size 5 laryngeal mask airway in males and females. AB - We compare the functional performance (ease of insertion, oropharyngeal leak pressure and anatomic position) of the size 5 LMA in males and females over a range of cuff volumes. We also determine 1) if age, height, weight and body mass index predict functional performance and 2) the relationship between oropharyngeal leak pressure (OLP) and anatomic position (judged by fibreoptic scoring). One hundred male and one hundred female paralysed, anaesthetised patients were studied. The number of insertion attempts and time to placement were recorded. OLP and fibreoptic score (FOS) were measured during inflation of the cuff from 0-40 ml in 10 ml increments. There were no failed insertions. Insertion times were similar for males and females. There were no differences in OLP or FOS between males and females at any cuff volume. For males, OLP increased from 0 to 10 ml and 10 to 20 ml (p < 0.001), but was unchanged from 20 to 30 and 30 to 40 ml. For females, OLP increased from 0 to 10 ml, 10 to 20 ml (p < 0.001) and decreased from 20 to 30 ml (p = 0.02), but was unchanged from 30 to 40 ml. For males, FOS increased from 0 to 10 ml (p = 0.02) and 10 to 20 ml (p < 0.001), but was unchanged from 20 to 30 and 30 to 40 ml. For females, FOS increased from 0 to 10 ml (p = 0.02), 10 to 20 ml (p < 0.001), decreased from 20 to 30 ml (p = 0.001), but was unchanged from 30 to 40 ml. There was a significant positive correlation with height and OLP in males and females at higher cuff volumes, but no correlation with any other variable. There was no correlation between OLP and FOS at any cuff volume for males, but there was a correlation at 0 (p = 0.03) and 10 ml (p = 0.01) for females. We conclude that ease of insertion, efficacy of seal and anatomic position for the size 5 LMA is similar in males and females. Age, weight and body mass index do not predict performance, but efficacy of seal improves with increasing patient height. Fiberoptically determined anatomic position is a poor predictor of efficacy of seal. The shape of the pharynx may be different between tall and short adults. PMID- 11050964 TI - [Anesthesia in vascular surgery. II]. PMID- 11050965 TI - [Unincorporated Channel Meeting (UCM)--basic research on mechanisms of anesthesia]. AB - Basic research on mechanisms of anesthesia in Germany has proved to be successful during the last years as demonstrated by a number of national and international publications and presentation. However, most of the groups working in this field lack a "crucial mass". In order to improve this situation the Unincorporated Channel Meeting (UCM) was introduced by Kochs and Urban in 1995. The meeting was established to provide a basis for the exchange of ideas and methods for all scientists interested in mechanisms of anaesthesia and not only anesthesiologists. Currently about ten groups are actively participating at the semi-annual meetings and the discussion group in the World Wide Web. The mini symposium was selected from papers presented at the March 1999 meeting in Hamburg/Germany and demonstrates the wide range of topics that were discussed including patch-clamp studies and whole animal research. The publication of this mini-symposium intends to provoke thoughts about the mechanisms of anaesthesia and to create interest in the scientific approach to these problems. PMID- 11050966 TI - [Effects of benzodiazepines and general anesthetics on the GABA A receptor: where are the differences?]. PMID- 11050967 TI - [Two-pore domain calcium channels]. PMID- 11050968 TI - [The non-competitive AMPA antagonist GYKI 52466 increases the anesthetic efficacy of methohexital]. PMID- 11050969 TI - [Ethanol specifically activates a Ca(2+)-dependent K(+) channel and reduces the firing frequency in sensory neurons]. PMID- 11050970 TI - [In vitro studies on muscular effects and side effects of anesthesiology relevant pharmacotherapy]. PMID- 11050971 TI - [Patch clamp studies on recombinant GABA A receptors using an ultra-quick application system]. PMID- 11050972 TI - [Autoradiographic evidence of RNA single strand breakages in brain tissue. An early marker of cerebral ischemia and hypoxia]. PMID- 11050973 TI - [Investigations on the binding of anesthetics to cerebral adrenoreceptors]. PMID- 11050974 TI - [Kinetics of inhibition of non-depolarizing muscle relaxants on the nicotinic acetylcholine receptor]. PMID- 11050975 TI - [Pneumatic external counterpulsation (PECP): a new treatment option in therapy refractory inner ear disorders?]. AB - BACKGROUND: The established therapeutic principle for sudden deafness and tinnitus is based on the stimulation of the inner ear perfusion by infusion therapy using vaso-active or hemodilutive agents. Concerning a mechanically induced increase of the inner ear's blood supply, the new technique of Pneumatic External Counterpulsation (PECP) was performed. This technique had already been used successfully in patients suffering from coronary heart disease. Due to an ECG-guided pneumatic compression of the lower extremities during diastole an increase of arterial perfusion of the extracranial supplying brain vessels can be obtained by PECP-treatment. PATIENTS AND METHODS: PECP was used in 33 patients (22 males, 11 females) between 19 and 76 years of age suffering from persisting acute hearing disorders and/or tinnitus after adequate infusion therapy. 3 patients revealed mild complications during PECP-treatment (e.g. thoracic pain) which seemed to be related to the therapeutic regime and disappeared completely after cessation of PECP. 30 patients underwent PECP-treatment on 5 following days for 1 hour. This standardized hourly treatment regime was extended to 10 days in 12 patients. Therapeutic effects were determined by color-coded duplexsonography and pure tone audiometry eventually masking the audiogram. RESULTS: During treatment an increase of 19% flow volume in the internal carotid arteries and of 11% in the vertebral arteries was evaluated using color-coded duplexsonography. In 47% of cases (n = 14) decrease of tinnitus intensity and/or tinnitus appearance with an average of 16 dB was perceived after the end of the treatment series. Hearing threshold increased in 28% of cases (n = 7) with an average threshold shift of 19 dB after PECP-treatment. All patients were examined by pure tone audiometry during the follow-up after 4, 8 and 12 weeks as well as after 6 and 12 months. In all cases the audiometric benefit lasted throughout the follow up 1 year, after treatment. CONCLUSIONS: The determination of a positive correlation between vascular flow increase in the extracranial brain-supplying vessels during PECP-treatment and the encouraging therapeutic results obtained by audiometry seem to make this new and promising therapeutic option effective, practicable and easy to handle. These preliminary results of the PECP technique should be validated in further studies featuring larger numbers of patients suffering from therapy-resistant inner ear disorders. PMID- 11050976 TI - [Main symptom: "pulse-synchronous tinnitus"]. AB - BACKGROUND: In comparison to cochlear or nerval generated ear noises, pulsatile tinnitus is a rare condition. Due to its own etiology, specific diagnostic steps are necessary. PATIENTS: We present 6 patients with pulsating tinnitus as the leading symptom. By means of these cases the various etiologies, rational diagnosis and therapy will be discussed. RESULTS: Pulsatile tinnitus is frequently caused by an increased blood flow in the cranial vessels through various pathologies. Besides those diseases going along with a general increase of blood circulation, regional alterations can be classified as hypervascular/hyperemic, arterial or venous conditioned. CONCLUSIONS: Physical examination and modern imaging can detect the underlying reasons in a quick and reliable way. PMID- 11050977 TI - [Distortion products of otoacoustic emissions (DPOAE) in acute tinnitus aurium]. AB - BACKGROUND: The origin of subjective tinnitus is not really known yet. It is possible to investigate the function of outer hair cells by means of the distortion products of otoacoustic emissions (DPOAE). METHODS: DPOAE of 56 patients were analysed in a prospective study (tinnitus without any loss of hearing n = 34; hearing loss with a maximum of 35 dB and tinnitus n = 22). There was always a normal tympanogram and in no case a conductive hearing loss. The relation of the primary tones f2/f1 was 1.22, the levels of the primary tones were 70 dB. RESULTS: DPOAE-amplitudes were reduced distinctly in both groups of patients. But there was no general difference and no difference at the frequency of tinnitus between the ear with tinnitus and the contralateral ear (no hearing loss) or there was a small reduction related to the normal ear (distinct hearing loss). DPOAE-amplitudes did not change in patients without hearing loss after a treatment with Pentoxifylline i.v., also not at the frequency of tinnitus, although there was a subjective reduction of tinnitus. In patients with hearing loss DPOAE-amplitudes enhanced like the improvement of hearing threshold at the frequency of tinnitus. CONCLUSIONS: The low DPOAE-amplitudes are a sign of damaged outer hair cells of the patients. But there is a symmetry of the DPOAE amplitudes and there is no change of DPOAE after a therapy with Pentoxifylline i.v. in patients without hearing loss. PMID- 11050978 TI - [Bilateral genuine cholesteatoma--a case report]. AB - We report on a patient who has been in otorhinolaryngological treatment for more than 24 years owing to recurrent infra-auricular fistulation. Early hospital admissions were due to suspected abscess-forming parotitis or lymphadenitis. As far as could be discerned, no sonographical or radiological diagnostics were conducted. Complications in the sense of meningitis or labyrinthitis were not reported. In the case of the aforementioned patient a bilateral anomaly in the area of the external auditory canal went hand in hand with the development of a bilateral genuine cholesteatoma. The auriculae were configurated normally. Corrective radical surgery was conducted on both sides, first the left, then the right side. What seems remarkable in this context is the fact that despite a over two decades old cholesteatoma there was no erosion of the osseous boundaries of the eardrum towards the labyrinth and cochlea and the osseous cover of the canalis facialis. The patient was diagnosed with a high degree loss of hearing. It was not possible to derive reproductive acoustic potentials intraoperatively, which pointed to deafness. Whether this deafness was congenital or developed in the course of time based on a growing sensory hearing loss could not be ascertained. PMID- 11050979 TI - [MR tomography study of the development of the sphenoid sinus]. AB - BACKGROUND: Aim of this study was to evaluate the postnatal growth pattern of the sphenoid sinus. PATIENTS: 83 cerebral MRI examinations of infants and children aged 5 months to 14 years were retrospectively reviewed for pneumatization and growth of sphenoid sinus. RESULTS: A continuous increase of pneumatization and growth of the sphenoid sinus was demonstrated between infancy and adolescence including considerable individual variations. Even in children less than two years old remarkable spatial extends of this sinus could be found in some cases. CONCLUSION: Diagnosis of an acute or chronical sinusitis in pediatric patients should alert the clinician to the possibility of a sphenoidal participation. PMID- 11050980 TI - [Endonasal endoscopic surgery of maxillary sinus mucoceles after Caldwell-Luc operation]. AB - BACKGROUND: Mucoceles of the maxillary sinus after Caldwell-Luc procedure develop by entrapment of mucosa. Specially mucoceles far laterally were being revised by a second transoral approach. This procedure carries the risk of damaging the infraorbital nerve. In addition the problem of scarring at the bottle neck between the lateral recessus and the nasal cavity with reclosure remains. METHODS AND PATIENTS: In 3 patients with a mucocele of the maxillary sinus 4-25 years after Caldwell-Luc operation an endonasal endoscopic opening of the mucocele was performed. In addition a silicon spacer was placed for 6 months. RESULTS: Twelve to fifteen months postoperatively all 3 patients still had an uneventful maxillary sinus according to the data of endoscopy, computer-tomographic or MRI examination. CONCLUSIONS: The endoscopic approach and the long standing placement of a splint seems to be a valuable alternative to a renewed somewhat risky Caldwell-Luc approach. PMID- 11050981 TI - [In vitro experiments with Nd:YAG laser surgery on the turbinates]. AB - BACKGROUND: Turbinate surgery is a therapeutic method for the treatment of the obstruction of nasal respiration. In this paper the dimensions of the laser lesions are described. In addition macro- and microscopical findings after laser surgery are given. METHODS: 10 human lower and 4 middle turbinates in vitro were treated with the Nd:YAG-laser in the non-contact mode (1064 nm, 2.5-25 W, cw). Stripe-like lesion with 3 cm length were produced. In addition the posterior end of the lower turbinates and the head of the middle turbinates were vaporized. RESULTS: Width, depth and volume of the lesions are given in dependence of laser power and irradiation time. The histological changes immediately after laser treatment are described. CONCLUSIONS: The energy doses for a clinical relevant stripe-like laser lesion of 3-4 cm in length of the lower turbinate is about 1500 Ws using Nd:YAG-laser. For evaporation of a posterior end of the lower turbinate 360 Ws are required using Nd:YAG-laser. For evaporation of the head of the middle turbinate a doses of about 1500 Ws are required using Nd:YAG-laser. PMID- 11050983 TI - [Excessive weight loss caused by dysphagia in superior laryngeal nerve neuralgia- a case report]. AB - BACKGROUND: Superior laryngeal nerve neuralgia is a very rare disease typically associated with paroxysmal and lancinating pain mostly localised on one side of the laryngeal region and sometimes radiating into the homolateral jaw or ear. The condition is usually idiopathic and treated successfully with carbamazepine. Even if the symptoms are unusual or not typical for superior laryngeal nerve neuralgia, the condition should be considered as a possible diagnosis if there are no other pathological findings. CASE REPORT: In October 1999, a 44-year old man presented a progressive dysphagia. Complaining about increasing problems in swallowing, he started refusing solid food and lost about 50 kg in two and a half years. In addition, he complained about persistent pain in the right laryngeal region, intensified by chewing or swallowing. Extensive investigations produced no pathological findings, especially no neoplastic process or other consuming diseases. After an injection of local anaesthetic near to the right hypothyroid membrane, the symptoms improved dramatically for a few hours and the diagnosis of superior laryngeal neuralgia was made. To confirm the diagnosis we injected sodium chloride as a placebo without any effect. The condition was successfully treated with 200 mg of carbamazepine three times daily without any major side effects. CONCLUSIONS: Even if the symptoms are not typical and dysphagia and weight loss are the outstanding symptoms, superior laryngeal nerve neuralgia should be considered as the possible diagnosis, especially if no other pathological findings are revealed. Injection of local anesthetic is a valuable tool for diagnosing the disease. PMID- 11050982 TI - [Dynamic imaging in diagnosis of dysphagia and globus sensation]. AB - BACKGROUND: Videofluoroscopy has gained high significance for the evaluation of deglutition disorders. Imaging alone cannot clarify whether the subjective symptom is represented morphologically or whether it is the substrate of a functional disorder. METHOD: The videofluoroscopies of 101 patients with dysphagia (n = 55) and globus pharyngitis (n = 46) were evaluated. Morphologic abnormalities were registered as well as sequential movement patterns. These data were compared with clinical and endoscopic findings. RESULTS: In 87% of the dysphagia and 74% of the globus patients videofluoroscopy revealed pathologic findings. Functional disorders were seen significantly more often than morphologic abnormalities. Highest incidence was found for cricopharyngeal dyskinesia (42%). Hypopharyngeal pouches and degeneration signs of the cervical spine with bolus impression less than 40% are common but functionally not important. Additional esophago-gastroduodenoscopy was pathological in 83% in the dysphagia group and 96% in the globus group. CONCLUSION: Videofluoroscopy is indispensable for the differential diagnosis of dysphagia and globus sensation, especially for the detection of functional disorders in the pharyngoesophageal segment thus documenting the dynamic aspect of deglutition. Videofluoroscopy should be completed by a gastroenterologic examination in order to improve diagnosis. PMID- 11050984 TI - [Reactivation of telomerase in squamous epithelial carcinomas in the area of the head and neck]. AB - BACKGROUND: Head and neck cancer development involves the accumulation of multiple cellular alterations over a long period of time. Selection and expansion of altered cell clones can lead to the evolution of a malignant phenotype. The theory of carcinogenesis suggests that unlimited cell proliferation is required for development of malignant disease and cancer must attain immortality for progression to malignant states. One step in the immortalization process may be the reactivation of telomerase. This enzyme complex can prevent the continuous shortening of telomeres which is observed at each cell cycle. RESULTS: Telomerase activity was detected in 68% of squamous cell carcinomas of pharynx and larynx and 58% of histologically tumor-free resection margins. Recurrences occurred with a higher rate in cases with telomerase positive primary tumor. The importance of telomerase activity in histologically negative resection margins needs further investigations. Telomerase activity was found in 90% of corresponding lymph nodes without any correlation to metastasis in the lymph node. CONCLUSIONS: The reactivation of telomerase seems to be an important step in carcinogenesis of head and neck cancers. Further studies are necessary in order to understand the role of the enzyme as a possible marker for tumor progression and clinical outcome. PMID- 11050985 TI - [An aberrant course of the internal carotid artery]. AB - Aberrant vascular courses of the A. carotis interna are extremely rare. They are usually combined with pulsatile symptoms. Missing symptoms are not proof of a non existing aberration, though. A paracentesis in such a situation leads to an initially unstoppable bleeding from the tympanion and tuba. For this reason it is necessary to contemplate a vascular reason for the local findings preoperatively. PMID- 11050986 TI - [Interesting case no. 38. Leiomyoma of the cervical esophagus]. PMID- 11050987 TI - [Plastic surgery reconstructions in the area of the neck. Superficial neck soft tissue--tracheostoma (1)]. PMID- 11050988 TI - Mycotoxin method evaluation. An introduction. PMID- 11050989 TI - Sampling techniques. PMID- 11050990 TI - Preparatory isolation of mycotoxins from solid phase fungal cultures. PMID- 11050993 TI - Measurement of aflatoxins using capillary electrophoresis. PMID- 11050991 TI - Preparation of mycotoxin standards. PMID- 11050994 TI - Liquid chromatographic method for aflatoxin M1 in milk. PMID- 11050992 TI - Electrospray mass spectrometry for mycotoxin detection and purity analysis. PMID- 11050995 TI - Immunochemical method for cyclopiazonic acid. PMID- 11050996 TI - Immunochemical method for ochratoxin A. PMID- 11050997 TI - Solution fluorometric method for deoxynivalenol in grains. PMID- 11050998 TI - Chromatographic methods for trichothecenes. PMID- 11050999 TI - Chromatographic method for the determination of the mycotoxin moniliformin in corn. PMID- 11051000 TI - Liquid chromatographic method for fumonisins in corn. PMID- 11051001 TI - Enzyme-linked immunosorbent assays of zearalenone using polyclonal, monoclonal and recombinant antibodies. PMID- 11051003 TI - Immunochemical method for citrinin. PMID- 11051004 TI - Solid phase extraction method for patulin in apple juice and unfiltered apple juice. PMID- 11051002 TI - Chromatographic method for Stachybotrys toxins. PMID- 11051005 TI - Liquid chromatographic method for the determination of ergot alkaloids in cereal grains. PMID- 11051006 TI - Chromatographic method for Alternaria toxins in apple juice. PMID- 11051008 TI - [Orthopedic pain therapy and "evidence-based medicine"]. PMID- 11051007 TI - [Shockwave treatment of femur head necrosis in the adult]. PMID- 11051010 TI - [A simple possibility for preventing treatment pain in ESWT of the locomotor system]. PMID- 11051011 TI - [Surgical fusion techniques of the lumbosacral transition zone]. PMID- 11051009 TI - [Robotics-assisted implantation of femoral components in hip endoprosthetics--an experimental study]. PMID- 11051012 TI - [Orthopedic care in Europe]. PMID- 11051013 TI - [Complete rotator cuff rupture--differential surgical techniques and intermediate term results]. AB - QUESTIONS: What is the mid term result of open rotator cuff repair? What are the criteria for a good surgical result? METHODS: Between February 1988 and December 1993 130 patients (135 shoulders) underwent surgery for rotator cuff repair. All surgical techniques were combined with a acromioplasty--transosseous refixation, transtendinous suture, isolated or combined transfers of tendons. The retrospective analysis consisted of clinical examination including the Constant score, X-rays and sonograms of the shoulder before and after surgery. RESULTS: 95 patients (98 shoulders) were controlled 4 years and 8 months after surgery by clinical, radiological and sonographical examination. 80% of all cases showed good or very good long term results by chiefly transosseous refixation alone or in combination with tendon transfer. Analysis of all data made it possible to create simple prognostic criteria. These criteria can help preoperatively to give an idea of the expected outcome. CONCLUSION: The best preoperative criteria for a prognostic good surgical result were an acromion-humeral head distance in the native true a.p.-X-ray of more than 7 mm, a rotator cuff defect of less than 2 x 3 cm and a passive free range of motion. PMID- 11051014 TI - [Secondary operations for improving shoulder function after brachial plexus lesion]. AB - The results of an integrated concept of therapy are presented including a description of indications and the various operative procedures to compensate insufficient shoulder muscles following brachial plexus lesion. PATIENTS AND METHODS: To improve stability and function of the shoulder in case of deltoid and supraspinatus paralysis 12 patients (1 female; 11 male; average age 29.4 years, range 17 to 56 years) underwent a shoulder arthrodesis. In 54 patients (11 female; 43 male; average age 30.3 years, range 18 to 69 years) a trapezius transfer was performed. The indication for a rotation osteotomy of the humerus to improve loss of external rotation due to paralytic infraspinatus muscle was determined in 4 male patients (average age 29.8 years, range 16 to 42 years). Our results are based upon an average follow-up of 2.0 (0.5-7.5) years after shoulder fusion, 1.9 (0.5-4.5) years after trapezius transfer and 1.6 (0.5-3.5) years after rotation osteotomy of the humerus. RESULTS: The trapezius transfer resulted in increased function of abduction of 6.2 degrees to 37.1 degrees (5 degrees-80 degrees) and forward flexion of 15.1 degrees to 36.2 degrees (10 degrees-90 degrees). A more stable condition of multidirectional shoulder instability was experienced by 50 patients (92.6%) and 49 patients (90.7%) were subjectively satisfied with the outcome of the operation. The strength and extent of functional improvement was, on average, greater following shoulder arthrodesis: abduction of 9.6 degrees to 65 degrees (40 degrees-90), forward flexion of 15.4 degrees to 59.2 degrees (30 degrees-90 degrees). 10 patients (83.3%) were subjectively satisfied with the outcome. Patients who had undergone external rotation osteotomy showed an average deficiency of external rotation of 20 degrees before operation. After osteotomy an improvement of 32.5 degrees to 12.5 degrees external rotation was achieved. All patients were satisfied with the increase of function. CONCLUSIONS: In patients with brachial plexus palsy, secondary operations according to the individual pattern of paralysis result in an improvement of shoulder function and stability as well as patients satisfaction. PMID- 11051015 TI - [Extracorporeal shockwave therapy in tendionosis calcarea of the rotator cuff: comparison of different treatment protocols]. AB - AIM OF THE STUDY: Extracorporeal shock wave therapy (ESWT) is a new therapeutic procedure for chronically painful calcifying tendinitis of the rotator cuff. The therapy may vary with the number of applied impulses or with impulse energy. Shock waves with an energy of 0.04 to 0.12 mJ/mm2 define low-dose ESWT, in contrast to high-dose ESWT (> 0.12 mJ/mm2). The aim of the study was to verify the hypothesis that either high-dose or low-dose ESWT could be effective if the total amount of applied energy was similar. METHOD: Fifty patients were assigned at random to 2 groups. The treatment consisted of 3 x 5000 low-dose impulses without anesthesia (group 1) and 1 x 5000 high-dose impulses with intravenous analgesia (group 2). The patients were examined at 6 weeks, 3 and 6 months after treatment. X-rays were performed at each visit. RESULTS: The Constant Score improved from 64.5 to 77.5 (group 1) and from 67.2 to 79.4 (group 2) before and 6 months after treatment (p < 0.05). The values on the visual analog scale which ranges from 0 (no pain) to 100 (maximal pain) improved from 76.8 to 48.8 (group 1) and from 75.4 to 45.6 (group 2) before and 6 months after treatment respectively. The final results for both Constant Score and visual analog scale were obtained after 3 months. X-rays showed a complete or subtotal calcific resorption in 8 (group 1, 32%) and 12 (group 2, 48%) patients. CONCLUSION: Extracorporeal shock wave therapy may be an alternative treatment of calcifying tendinitis of the shoulder. Both treatment protocols gave equivalent results. PMID- 11051016 TI - [Comparison of results of results of medium energy ESWT and Mittelmeier surgical therapy in therapy refractory epicondylitis humeri radialis]. AB - INTRODUCTION: During the last years the extracorporal shock wave therapy has shown its efficiency in the treatment of the conservative non curable epicondylitis humeri radialis. This study evaluates the outcome of the ESWT and the operative treatment. METHODS: Between 1996 and 1997 60 patients suffering from lateral epicondylitis were analyzed. 30 were treated operative by using the Mittelmeier-procedure (tangential partial bone resection of the epicondylus humeri radialis) and further 30 with extracorporal shock waves. These patients received 2000 impulses of 0.23 mJ/mm2 in two sessions. RESULTS: Follow-up of the group who underwent the Mittelmeier procedure showed good and excellent results in 73% after one year using the score of Roles and Maudsley (1972). The patients treated by the ESWT showed good and excellent results in 43%. DISCUSSION: Regarding to the duration of symptoms and large scale of the primary treatment and operative hazards the results are showing the benefit of the medium energetic extracorporal shock wave application in the treatment of patients with no response on regular therapy. PMID- 11051017 TI - [Effect of patient age on use of the knee joint score]. AB - AIM OF THE STUDY: Clinical studies are done with the help of scores though different factors of influence lower comparability. The underlying study examines the influence of patient age as this always presents a certain span. METHOD: 96 degree I damaged knee joints were examined by 3 examiners using the Larson-, Lysholm-, Marshall-, HSS- and OAK-score. Furthermore ratings by a VAS and the Tegner activity score were done. With the Friedman test, the rank correlation coefficient by Spearman and the contingency chart by Bowker it was tested if the examiners and the scores rate equally. To find out the influence of the age three age groups were made up. RESULTS: The examiners judged significantly different excluding the Marshall and the OAK scores. In between two examiners no significant difference could be found between the young and the middle-aged patient group. Between two other examiners no significant difference was found only for the Lysholm and the HSS score in the young group and only for the Lysholm score in the middle-aged group. In the group of the senior patients no significant difference for the scores by Lysholm, Marshall and OAK were found. In the comparison of the second examiner pair no significant differences could be proven by the scores by Larson, the OAK and the HSS. All other comparisons were significantly different. In the comparison of all five scores significant differences were seen between the Larson score and the HSS and the Lysholm and the HSS. The Lysholm score proved to be the strictest, the HSS to be the leanest. CONCLUSION: Especially for the senior patients in dependency of the examiner and the chosen score significant differences were found concluding that the relevance of results lessens for future times as the score results drop with patient age anyway. The age span in one study should have a maximum of 10-20 years to reduce the influence of age on the final result. PMID- 11051018 TI - [Significance of sagittal stability in knee prosthesis implantation--an analysis of 76 cases with unconstrained joint surface replacement]. AB - ISSUE: How does the sagittal stability influence the outcome in unconstrained knee arthroplasty? METHOD: In order to clarify this aspect, 76 arthroplasties (10 male, 66 female, 39x gonarthrosis, 37x rheumatoid arthritis) in 61 patients with unconstrained primary knee arthroplasty were examined with a mean follow-up of 4 years. The determined values were the HSS-Score, the Knee-Society-Score, the range of motion, the flexion contracture as well as the posterior and anterior drawer with the KT 1000. The laxity was defined as the sum of the anterior and posterior drawer. RESULTS: The mean values measured were 2.9 mm for the anterior drawer, 1.9 mm for the posterior drawer and 4.8 mm for the laxity. The total patient population reached 81.3 points in the Knee Score, 70.9 points in the Function-Score and 80.7 points in the HSS-Score. The medium range of motion was determined as 103.5 degrees, the medium flexion contracture as 3.5 degrees. For an anterior drawer of > 6 mm and a posterior drawer of < 1 mm the results deteriorated significantly. A laxity of 8-11 mm gave the best score results. CONCLUSION: An anterior drawer of < 6 mm, a posterior drawer of 2-5 mm and a laxity of 8-11 mm seem to be recommendable for unconstrained knee arthroplasty. PMID- 11051019 TI - [Results of surgical arthrolysis in treatment of restricted knee joint movement]. AB - PROBLEM: The study's task is to describe the results of the 58 arthrolysis of the knee joint, which were performed between 1967 and 1995. Additionally, we wanted to find out about the influencing factors on the long-term results after this operation. METHOD: The data used for this retrospective study is based on the complete charts, in which the findings of the postoperative follow-up examinations were documented (range of motion, local findings, gait). The average follow-up time was 3.2 +/- 3.6 years. In order to draw conclusions, we focused on the extent of motion and on the relative benefit of motion. Despite of the inhomogeneity of the collective of patients, we performed statistical tests for a better presentation. RESULTS: The mean knee joint moveability was increased from 0 degree/11 degrees/46 degrees to 0 degree/7 degrees/90 degrees at the last follow-up date. The relative benefit was 44.5% (Median 54%). Additionally, the extraarticular procedures had the best, the combined arthrolysis the worst outcome. Postinfectious stiffness of the knee did not show a worse prognosis in comparison to non-infectious causes for a decrease in motion. CONCLUSIONS: Depending on the preoperative findings, the range of moveability was considerably better, when the indication for the performance of an arthrolysis was recognized in time. PMID- 11051020 TI - [Spinal contusion after trauma to the cervical spine--relevance of the sagittal diameter of the spinal canal]. AB - PURPOSE: The purpose of this study was to analyze the relationship between spinal stenosis and cervical myelopathy. Furthermore it was to investigate whether there is a correlation between spinal stenosis and progression of neurologic symptoms? PATIENTS AND METHODS: Using the Pavlov ratio method, we compared 14 patients with a contusio spinalis with 14 patients who suffered from a distortion of the cervical spine. This ratio is calculated by the sagittal diameter of the spinal canal with the anteroposterior width of the vertebral body. RESULTS: In eight patients with a contusio spinalis, there was a spinal stenosis, while there was none in the distortion group. The remaining patients with a contusio had severe degenerative changes in routine roentgenograms of the cervical spine. The Pavlov ratio was statistically significant reduced in the contusio group. After 8.5 days there was a complete regression of neurologic symptoms, whereas in 4 patients there was no full recovery. CONCLUSION: To conclude we demonstrated that a spinal stenosis promotes a contusio spinalis but there is no evidence that the Pavlov ratio allows statements concerning the progression of the injury. PMID- 11051021 TI - [Ependymoma of the thoracic spine. Cervico-brachialgia as the first symptom--a case report]. AB - Most frequently the ependymomas of the central nervous system affect the Conus medullaris and Filum terminale. Ependymomas of the thoracic spine with association of a tumor-caused syringomyelia are extremely rare. The reported 50 years old patient with a thoracic ependymoma realized first symptoms as a shoulder-arm pain caused by the tumor-syrinx. The case report indicates that MRI is the superior imaging modality for primary tumors of the central nervous system. All relevant diagnostic and therapeutic informations derived from the MRI. The article describes the clinical manifestations, the diagnostic way and therapeutic procedure in the problem region thoracic spine. PMID- 11051023 TI - [Examination of Heberden arthrosis with a histological-histochemical score]. AB - QUESTIONS: In case of Heberden's nodes the osteoarthritis starts with a so called tidemark flaking as a reaction to a subchondral ossification. The question was, if a differentiation in relation to a control group by an objective score is possible. METHODS: 218 finger joints from 56 cadavers were investigated morphologically (score). RESULTS: The osteoarthritis starts with a subchondral ossification. At this time the surface of the cartilage is not destroyed yet. Reactive tidemark flaking is the beginning of the general degradation. In relation to a control group a significant differentiation by a histological score is possible. CONCLUSIONS: In case of Heberden's nodes the osteoarthritis starts with the subchondral ossification. Because Heberden's nodes are the specific manifestation of the Generalized Osteoarthritis on the distal finger joints, further studies have to assess, if the same pathogenetic mechanism can be seen in osteoarthritis of the large joints. PMID- 11051022 TI - [Correlation between degenerative changes of the thoracic spine and the finger joints--a roentgenological study in generalized osteoarthrosis]. AB - PURPOSE: Different studies in patients with rheumatoid arthritis demonstrated a correlation between degenerative changes of the cervical spine and peripheral joints. The aim of study therefore was to assess the relations of degenerative changes of the cervical spine and peripheral joints. METHODS: 106 patients suffering on Generalized Osteoarthritis were examined by standard radiography of the hands and cervical spine. All images were evaluated according to Kellgren grades. The mean values of the cervical spine and finger joints were compared in three calculations, for this purpose the finger joints were "segmental", "functional" and randomized arranged. RESULTS AND CONCLUSIONS: Concerning quality and quantity the highest correlation was observed in the group of the so-called "segmental-arrangement". Therefore a association between the cervical spine and the finger joints seems possible relating to manifestation, pattern and graduation of degenerative processes. At this stage there is no statement possible about the direction of the correlation. PMID- 11051024 TI - ["Lumbar intervertebral disk-induced sciatica" diagnostic error in extensive extra- and intrapelvic lipoma]. AB - PURPOSE: The aim of this study is to demonstrate the importance of the differential diagnosis of sciatica. Radiculopathy in the lower extremity of an adult usually originates from a herniated nucleus pulposus. In this paper an extraspinal cause of sciatica due to a pelvic tumor is reported, which is initially often not recognized. METHODS AND RESULTS: A 67-year old woman suffered from sciatica. After one year of unsuccessful conservative treatment and beginning weakness of hip flexion a computed tomography of the pelvis revealed a giant lipoma with compression of the lumbosacral plexus. CONCLUSIONS: The results of this case illustrate the problem of over-rating CT-findings of the lumbar intervertebral disks in patients with sciatica. Extraspinal tumorous causes of radiculopathy are rare, but should be considered if the therapeutical measures are resistant to treatment. Computed tomography and magnetic resonance imaging are most useful in confirming a retroperitoneal tumor causing lumbosacral radiculopathy. PMID- 11051025 TI - [Gluteal compartment syndrome after intramuscular gluteal injection]. AB - Complications after i.m. injection are rare. Only few cases need emergency operative treatment. This case report shows the exclusive situation of a gluteal compartment syndrome caused by a hematoma. Clinical findings showed signs of nerve compression with sciatic pain. Only immediate surgical treatment prevented persistent nerve or muscle tissue damage. PMID- 11051026 TI - [Pathological rupture of the distal biceps tendon after long-term androgen substitution]. AB - QUESTION: Is the long-standing use of androgenes able to cause tendon lesions with pathological tendon ruptures? METHOD: In a case of a rupture of the distal biceps tendon after long-standing testosterone-substitution it is tried to show the connection between the use of androgenes and pathological tendon rupture by the help of the patients treatment documents, the X-rays and sonograms, the histological findings and the results of the clinical examination. RESULTS: For the first time a case with a rupture of the distal biceps tendon after long standing testosterone-substitution for the support of a genital transformation is described. Since other tendon damaging factors could be excluded the suspicion of tendon alteration caused by androgenes is obvious. CONCLUSION: Looking at tendon ruptures of professionals and even amateur sportsmen the possible connection between long-standing use of androgenes and tendon damage has to be considered. PMID- 11051027 TI - [Cerebellar hemorrhage as an early complication of spinal operations. 2. Case reports and review of the literature]. AB - GOAL: The present paper investigates the etiology and pathogenesis of cerebellar hemorrhage after spine surgery. METHOD: This paper reports two patients in those this complication was seen. In respect to the current literature we discuss the etiology and pathogenesis of cerebellar hemorrhage due to spine surgery. RESULTS: Cerebellar hemorrhages represent a life-threatening situation. There are no reports in the literature about cerebellar hemorrhage as an early complication of intraoperative dura injuries in spine surgery. It seems that a bigger cerebrospinal fluid loss is responsible for the developing of cerebellar hemorrhages. The loss creates a pressure gradient from infratentoriell to site of lesion and also leads to mechanical stress on cerebellar blood vessels such as traction, tearing and kinking. CONCLUSIONS: Every condition after spine surgery with dura injuries and neurological deficits should be carefully evaluated and intracranial hypotension as well as hemorrhage should be ruled out. PMID- 11051028 TI - [Synovial hemangioma within Hoffa's fat pad as the cause of anterior knee pain. 2 cases within a family and review of the literature]. AB - An 18 years old man and his younger sister suffered from anterior knee pain. Cause is a soft tissue tumor located within Hoffa's fat pad however in both cases. The tumor has been excised and the histological examination demonstrated the diagnosis synovial hemangioma. A literature search shows that synovial hemangiomas are rare soft tissue tumors. They mainly involve the knee joint. Mean age is the second decade of life and most authors observed severe degenerative changes in the involved knee joint. There are no reports about familiar accumulation within the literature. Familiar accumulation of synovial hemangiomas however have been described for cerebral, skin and hepatic manifestations but not for synovial hemangiomas. PMID- 11051030 TI - The careful correction of renal insufficiency abnormalities: early is good. AB - Cardiovascular disease in end-stage renal disease (ESRD) patients starts early in the course of renal insufficiency and may be aggravated by renal replacement therapy. Many potential reversible risk factors are present in renal insufficiency. The identification and correction of these risk factors, in part delaying progression of renal failure, may reduce their role in ESRD cardiovascular disease significantly. Early referral of renal insufficiency patients to a nephrologist may be the real answer to the cardiovascular morbidity and mortality of ESRD patients. PMID- 11051029 TI - [Comment on Breusch, S. J., Y. Draenert, K. Draenert: The anatomical basis of the cemented femoral stem. A comparative study of straight and anatomical design]. PMID- 11051031 TI - AGEs and carbonyl stress: potential pathogenetic factors of long-term uraemic complications. AB - Advanced glycation end products (AGEs) may be involved in the pathogenesis of complications of chronic renal failure. Pentosidine, a carbohydrate-derived AGE, is considerably elevated in uraemic patients. This compound per se has no biological activities but is highly correlated to the levels of precursors of carbonyl compounds, and for this reason is considered a reliable surrogate marker for AGEs. The modification of proteins in uraemia is not limited to AGEs, since advanced lipoxidation end products are also demonstrable in plasma proteins in uraemia. The accumulation of these compounds does not seem to be dependent only on the decline of renal function. Carbonyl precursors of AGEs and advanced lipoxidation end products are markedly elevated in uraemic patients. On this basis, the 'carbonyl stress' theory has been formulated. This theory holds that increased oxidation of carbohydrates and lipids and/or inadequate detoxification of carbonyl compounds may contribute to long-term complications of end-stage renal disease such as dialysis amyloidosis and cardiovascular diseases. Preliminary cross-sectional studies in haemodialysis patients seem to indicate that the AGEs and carbonyl stress may be involved in the pathogenesis of alterations in left ventricular geometry and function in these patients. PMID- 11051032 TI - Membranes, technologies and long-term results in chronic haemodialysis. PMID- 11051033 TI - Clinical implications of biocompatibility in blood purification membranes. PMID- 11051034 TI - Microbiological purity of dialysate for on-line substitution fluid preparation. AB - Dialysate purity has become a major concern in recent years since it was shown that low levels of endotoxin in dialysate were able to induce the production of proinflammatory cytokines, which were putatively implicated in the development of dialysis-related pathology. On-line haemodiafiltration (HDF; or haemofiltration) using the dialysate as the source of substitution fluid magnifies this risk and reinforces the critical role of the dialysate quality to be used. In order to virtually abolish the risk related to dialysate contaminants, it is mandatory to ensure the highest purity of the dialysate used in order that the substitution fluid produced satisfies the quality demands of a sterile and pyrogen-free infusion solution. Ultrapure dialysate production is therefore a common need for all on-line systems where substitution fluid is prepared continuously by sterilizing filtration of the dialysate. However, since dialysate purity plays a role in the complex haemocompatibility interaction which occurs during the haemodialysis session, the use of ultrapure dialysate must be considered as a suitable option for all haemodialysis modalities. To achieve this goal, one must keep in mind that ultrapure dialysate and infusate result from a complex chain of production where ultrapurity and/or sterility of the final solution relies on the weakest or worst component of the chain. Reliable production of ultrapure dialysate and infusate relies on several prerequisites: use of ultrapure water, use of clean electrolytic concentrates, implementation of ultrafilters on specifically designed HDF machines, microbiological monitoring of the chain with adequate and sensitive methods, and hygienic handling of the chain including frequent disinfection to reduce the level of contamination and to prevent biofilm formation. When properly done, the safety and reliability of on-line systems have been confirmed in large clinical studies. It is now time to validate the on-line process in large controlled clinical trials. PMID- 11051035 TI - Dialysis membranes in convective treatments. PMID- 11051036 TI - Removal of small and middle molecules by convective techniques. AB - The current renewed interest in haemofiltration (HF) is due mainly to its potential advantages in reducing morbidity and mortality. We analysed the data obtained from eight patients treated with pre-dilution HF and compared them with the calculated post-dilution HF and haemodialysis (HD) data in order to quantify the depurative performance of HF and to assess whether its claimed ability to improve cardiovascular stability is at least partially related to less sodium removal. The urea clearance (as a marker of small molecules) of pre-dilution HF was equivalent to that of standard low-flux HD and approximately 20% higher than that of postdilution HF. For a calculated increase of 117% in the dialytic clearance of beta2-microglobulin using high-flux HD, the increase in beta2 microglobulin removal with HF was only 30%. The intradialytic decrease in plasma water beta2-microglobulin concentrations was 73% (vs 53% in HD) followed by a post-dialytic rebound of 85% (vs 42% in HD). The interdialytic concentrations of beta2-microglobulin were always lower. At the start of the subsequent session, the beta2-microglobulin concentrations observed in HF-treated patients were 92% of those in the patients treated with HD. Sodium removal was similar using both HF modalities at the same total dialysate and reinfusate sodium concentrations, and lower than in the case of HD. HF makes it possible to obtain constantly lower plasma water beta2-microglobulin concentrations whose long-term clinical relevance has to be verified. Under usual operational conditions, sodium removal is less with pre-dilution HF than with HD, thus raising some doubts about the intrinsic capacity of HF to improve cardiovascular stability by a mechanism other than less sodium removal. PMID- 11051037 TI - Acid-base correction and convective dialysis therapies. AB - Whichever dialysis therapy is used, there is a similar need for correcting the acid base balance. The most important tool for this is the buffer in the dialysis fluid and, when using convective therapies, also in the substitution solution. The buffer source in all modern versions of these therapies should be bicarbonate. The more efficient the dialysis treatment in terms of small solute transport, the more rapid the uptake of buffer. Thus, optimally applied haemodiafiltration has the potential for the largest buffer gain. The target for acid-base correction in dialysis is to maintain patients within or as close to the physiological plasma bicarbonate range as possible. However, cross-sectional studies of acid base status among patients treated with contemporary forms of dialysis often show moderate acidosis. As metabolic acidosis has been found to be an important stimulus for protein catabolism in experimental studies, an association with nutritional problems has been sought in dialysis patients. This has revealed a negative correlation between plasma bicarbonate and nutritional parameters. Acidotic patients were found to have better nutritional status than patients with normalized acid-base balance. However, caution should be exercised when interpreting plasma bicarbonate levels, since acidosis may be a cause as well as an effect of excessive protein catabolism. Although available clinical data suggest that the catabolic effect of mild acidosis can be compensated by adequate nutrition and adequate dialysis, it should be desirable to aim for a normalized acid-base balance in combination with adequate nutritional intake and delivery of dialysis. PMID- 11051038 TI - The impact of haemofiltration on the systemic cardiovascular response. AB - Convective transport across the dialysis membrane has long been known to be a good alternative to diffusion. Among the advantages of haemofiltration over conventional haemodialysis is a better cardiovascular stability. Haemofiltration has a more physiological response to fluid removal than haemodialysis: (i) blood volume may be better preserved, especially in patients with a compromised cardiac function; (ii) arterial peripheral resistances and venous tone increase; and (iii) myocardial contractility is neither depressed nor altered. We discuss the role of the different pathophysiological mechanisms in the disparity in cardiovascular reactivity between haemofiltration and haemodialysis. PMID- 11051039 TI - Pre-dilution haemofiltration--the Sardinian multicentre studies: present and future. The Sardinian Collaborative Study Group on Haemofiltration On-Line. PMID- 11051040 TI - Adequacy in pre-dilution haemofiltration: Kt/V or infusion volume? The Sardinian Collaborative Study Group on Haemofiltration On-line. AB - Kt/V is the main index of adequacy for diffusive and diffusive convective methods of extracorporeal depuration, yet there exists no universally acceptable validation of an adequacy index for the solely convective methods such as haemofiltration (HF). The aim of the present study is to analyse which of the parameters of adequacy used in two multicentre HF studies, Kt/V for urea or infusion volume, correlate best with nutritional parameters and can therefore be utilized for the evaluation of treatment dose in on-line pre-dilution HF. Twenty three clinically stable patients were enrolled in the first study [3 months of haemodialysis (HD)+ 3 months of HF]. In the second study, 24 stable patients were studied in three phases: 6 months in HF, 6 months in HD and a further 6 months in HF; in this study, a target of Kt/V= 1.2 in all three periods was preestablished: 15 patients completed the full study. In both studies, we utilized the same monitor (AK 100/200 Ultra, Gambro), the same membrane (polyamide) and the same on line prepared ultrapure dialysis fluid and sterile infusion solution. In both studies, we ensured that HF fulfilled the following parameters of adequacy: urea kinetics, cardiovascular and blood pressure stability (better in HF than in HD), common haematochemical and nutritional parameters, reduction in beta2 microglobulin levels, a good intra- and extra-session clinical outcome, and a good quality of life with morbidity and mortality rates no different from those of HD. HF proved to be an efficacious method of ensuring adequate depuration and a good quality of life for uraemic patients. We have shown that in longer periods of HF, a notable correlation between Kt/V and normalized protein catabolic rate (nPCR) and an equally good correlation between total ultrafiltration (UF)/dry weight ratio and nPCR could be achieved. In both studies, the patients showed a good level of epuration adequacy when total UF per session was at least 1.3 times the dry body weight. The total UF/body weight ratio thus seems to be an easy method in HF because of its greater ease of predictability and measurement, also when it is used independently of the Kt/V index. PMID- 11051041 TI - Molecular markers for predicting response to tamoxifen in breast cancer patients. AB - Tamoxifen is one of the most effective treatments for breast cancer. Standard practice is to select patients who are likely to respond to this therapy through the evaluation of estrogen receptor (ER) and progesterone receptor (PR) in the primary tumor tissue. Over the past 25 yr that physicians have been using ER determination to guide tamoxifen use, numerous studies have demonstrated that this molecular marker is useful in predicting benefit from tamoxifen. ER has been analyzed for many years using ligand-binding assays. However, current practice involves the use of immunohistochemical-based assays to detect ERalpha Immunohistochemistry (IHC) has several advantages. For example, IHC evaluates tumor cell heterogeneity, can be used to study small samples, is less expensive, and allows direct correlation with multiple histopathological tumor features and other molecular markers. PR, an estrogen-responsive protein, can also be useful in predicting response to tamoxifen in specific clinical situations. In recent years, several other markers of tamoxifen response have been examined, including: pS2 (another estrogen-regulated protein), heat-shock proteins 27 and 70, bcl-2 protein, c-erbB-2 (HER-2/neu) oncoprotein, and mutated p53 tumor suppressor protein. In this article, we present an analysis of the data on these new molecular markers. Overall, from numerous studies, the data indicate that in addition to ERalpha bcl-2 is a potential candidate to help further improve our ability to predict response to tamoxifen. ER and bcl-2 are the most useful molecular markers to better identify breast cancer patients who will respond to tamoxifen and who will have prolonged survival. PMID- 11051042 TI - Interleukin-6 regulation of kappa opioid receptor gene expression in primary sertoli cells. AB - Three classes of opioid receptors--mu, delta, and kappa--mediate physiological and pharmacological functions of the endogenous opioid peptides and exogenous opioid compounds in the central nervous system (CNS), as well as in peripheral tissues including the immune system. Using reverse transcriptase polymerase chain reaction (RT-PCR) analysis, we show that freshly isolated and highly purified somatic (Sertoli and Leydig) and specific germ (spermatogonia, pachytene spermatocytes, round, and elongating spermatids) cells of the rat testis differentially express the mRNAs for these opioid receptor genes. Furthermore, to identify a functional mechanism for cytokine regulation of testicular opioid receptor gene expression, we employed primary Sertoli cells as a model system. In a semiquantitative PCR analysis using the S16 ribosomal RNA gene as an internal control, we show that interleukin-6 reduces kappa opioid receptor mRNA levels from 6 to 24 h of treatment in primary Sertoli cells. This regulation requires new RNA and protein synthesis and is partially mediated by the protein kinase A pathway. These findings are consistent with a role for the cytokine and opioid signaling pathways in Sertoli cellular function and the interaction that exists between the opioid and the immune systems in the CNS. PMID- 11051043 TI - Effects of labor on pituitary expression of proopiomelanocortin, prohormone convertase (PC)-1, PC-2, and glucocorticoid receptor mRNA in fetal sheep. AB - We hypothesized that the concurrent prepartum rise in adrenocorticotropic hormone (ACTH) and cortisol in the plasma of fetal sheep might be attributable to altered expression of pituitary endoproteases, prohormone convertase (PC)-1, and PC-2, or to changes in pituitary expression of glucocorticoid receptor (GR) that would influence negative feedback potential. We obtained pituitary tissue from fetal sheep during late pregnancy (d 100-d 145, term) and at precise times during the process of labor and used in situ hybridization to localize and quantify mRNA levels. Proopiomelanocortin (POMC) mRNA was regionally distributed (pars intermedia > inferior pars distalis > superior pars distalis) and increased within the pars distalis during late pregnancy and with labor. At term, levels of PC-1 and PC-2 mRNA were higher in the pars intermedia than pars distalis; PC-1 but not PC-2 in the pars distalis increased with gestational age, although it did not change further at labor. GR mRNA levels in the pars distalis increased between d 135 and term, then decreased during labor. We suggest that the concomitant rise in plasma ACTH and cortisol of fetal sheep during late gestation may be attributable, in part, to increased expression of PC-1 leading to increased POMC processing. Furthermore, the negative feedback effects of cortisol on pituitary POMC synthesis and/or ACTH release during active parturition may be lessened by downregulation of anterior pituitary GR. PMID- 11051044 TI - Ovarian influence on gonadotropin and prolactin release in mated rabbits. AB - In 17beta-estradiol (E)-treated ovariectomized (OVX) rabbits, the coitus-induced luteinizing hormone (LH) surge is only one fourth that in ovarian-intact rabbits. In this study, we determined the pattern of the coitus-induced gonadotropin release, i.e., LH and follicle-stimulating hormone (FSH), in OVX + E animals without or with continuous 3-wk treatment of 20-alphahydroxypregn-4-en-3-one (20alphaP). For positive and negative experimental controls, ovarian-intact rabbits were either mated or sham mated, respectively. The pituitary hormones prolactin (PRL) and growth hormone (GH) were measured to serve as collateral controls for gonadotropins. The addition of continuous 20alphaP in OVX + E does fail to stimulate a coitus-induced LH surge equal in magnitude and duration to the LH surge in ovarian-intact rabbits. Postcoital levels of FSH were greater in OVX + E + 20alphaP animals than those in OVX + E rabbits. Coitus induced a PRL surge in ovarian-intact and OVX + steroid-treated females, but not in mated males, thereby suggesting a gender difference in this neuroendocrine circuit. Neither coitus nor steroids altered plasma GH values in female or male animals. We conclude that chronic administration of neither E nor E + 20alphaP can restore full-scale gonadotropin surges in OVX rabbits, whereas replacement of one or both of these steroids is sufficient for a coitus-induced PRL surge. Moreover, the presented observation that activin stimulates hypothalamic gonadotropin-releasing hormone (GnRH) release suggests a possible involvement of ovarian proteins in the production of a full-scale coitus-induced GnRH/LH surge. PMID- 11051046 TI - Homologous upregulation of gonadotropin-releasing hormone receptor mRNA occurs through transcriptional activation rather than modulation of mRNA stability. AB - In a previous study, we showed that even continuous application of gonadotropin releasing hormone (GnRH) could increase the steady-state levels of GnRH receptor (GnRH-R) mRNA if treated for a relatively short period (6 h). Therefore, in the present study we examined whether GnRH-induced increment of GnRH-R mRNA is owing to stabilization of the preexisting GnRH-R mRNA or new synthesis of GnRH-R mRNA or both. Initially, to examine the effect on new RNA synthesis, the transcription inhibitor, actinomycin D (2 microM), was added to primary cultured rat anterior pituitary cells. In the presence of transcription inhibitor, GnRH-induced augmentation of GnRH-R mRNA levels was completely abolished. This result indicates that homologous upregulation of GnRH-R mRNA expression occurs at least through new RNA synthesis of GnRH-R gene. We further assessed the effects of GnRH on the turnover rate of GnRH-R mRNA using actinomycin D (2 microM). The basal half-life of GnRH-R mRNA was estimated to be approx 21 h. The application of GnRH tended to slightly suppress the basal turnover rate of GnRH; however, there was no statistically significant difference, compared with the group treated with actinomycin D alone. Collectively, our results suggest that the homologous upregulation of GnRH-R mRNA may occur through transcriptional activation of GnRH R gene rather than enhancement of GnRH-R mRNA stability, although we did not examine the transcription rate of GnRH-R gene. PMID- 11051045 TI - Structural and functional effects of high prolactin levels on injured endothelial cells: evidence for an endothelial prolactin receptor. AB - Stress has been linked to health problems such as atherosclerosis and prolonged wound healing, which involve the responses of injured endothelial cells. Though prolactin (PRL) levels become increased during the physiological response to stress, the significance and effects of these increases are largely unknown. Here we examined the effects of elevated, though physiological, concentrations of PRL on the responses of cultured endothelial cells after mechanical injury to cell monolayers. When treated at the time of injury with PRL levels of 62.5-1000 ng/mL, cells at the wound front became abnormal in shape and had reductions in f actin staining in comparison to controls that were not PRL-treated. High PRL concentrations also inhibited the adhesion of cells to their growth surface in a dose-dependent manner. Using rhodamine-labeled PRL, we observed specific PRL uptake by these cells that suggested the presence of a PRL receptor. Finally, mRNA for the long form of the PRL receptor was detected by RT-PCR. To our knowledge, this is the first report demonstrating that (1) high PRL concentrations alter the actin cytoskeleton and adhesion of injured endothelial cells and (2) endothelial cells express the transcript for the PRL receptor. Thus, we report novel effects of PRL that may be mediated by activation of an endothelial cell PRL receptor. PMID- 11051047 TI - c-Jun targets amino terminus of androgen receptor in regulating androgen responsive transcription. AB - The human androgen receptor (hAR) is a member of the nuclear receptor superfamily and functions as a ligand-inducible transcription factor. We have previously proposed that c-Jun mediates the transcriptional activity of this receptor. The modular nature of hAR was used in this study to generate several fusions with the heterologous DNA-binding domain of the yeast transcription factor GAL4 in an attempt to identify the c-Jun-responsive domains within the receptor. Our results suggest that the target of c-Jun action is the amino terminus (AB region) of the receptor and that hAR amino acids 502-521 are critical for the c-Jun response. Additionally, amino acids 503-555 were shown to harbor an autonomous transactivation that is stimulated by c-Jun. Furthermore, we demonstrated that transcription intermediary factor-2 (TIF-2), a coactivator that acts on the activation function-2, stimulates the full-length hAR. These results suggest that c-Jun and TIF-2 can work together as coactivators on the hAR by targeting distinct portions of the receptor. PMID- 11051048 TI - Functional interactions between the extracellular domain and the seven transmembrane domain in Ca2+ receptor activation. AB - We studied the activity of mutants involving the aminoterminal extracellular, seven-transmembrane (7TM) and carboxy-terminal tail domains of the human Ca2+ receptor to gain insight into the functional interactions between these domains during receptor activation. Missense mutations of highly conserved residues, D190 and E297, in the extracellular domain (ECD), and a mutation within part of the proximal carboxyterminal tail, A877-880E, resulted in receptors with severely reduced response to Ca2+ despite adequate cell surface expression. Coexpression of either D190A or E297K mutants with A877-880E led to significant reconstitution of function. No such reconstitution occurred when D190A or E297K mutants were coexpressed with a truncation mutant possessing an intact amino-terminal extracellular and first transmembrane domain, despite evidence for heterodimerization and cell surface expression of the respective mutant receptors. In addition, no reconstitution of function was observed when D190A was coexpressed with a deletion Ca2+ receptor mutant lacking only a cysteine-rich region located in the ECD of the Ca2+ receptor (Ca-//-Ca). Moreover, coexpression of this Ca-//-Ca with A877-880E did not recover function. The results show that Ca2+ receptor extracellular and 7TM domains are discrete entities that can communicate within the context of a heterodimer composed of complementary mutant receptors. Two intact 7TM domains and two intact cysteine-rich regions appear to be required for such communication to occur. The results are discussed in the context of a speculative model of receptor structure and function. PMID- 11051049 TI - Insulin-like growth factor-1 causes a switch-like reduction of endogenous growth hormone mRNA in rat MtT/S somatotroph cells. AB - Reduction of mRNA expression from the endogenous GH gene by insulin-like growth factor 1 (IGF-1) in somatotroph-like rat MtT/S cells was measured. GH mRNA levels were reduced by 65 nM IGF-1 treatment in a time-dependent manner over 5 d of culture with a calculated GH mRNA half-life of 50 h, in line with previous values from primary cultures. Inhibition of inositol 3-phosphate kinase by wortmannin or LY-294,002 treatment was ineffective in blocking IGF-1 decreases in GH mRNA, as was inhibition of MAP kinase activity by PD 098059. The inhibition by IGF-1 also did not regulate Pit-1 (GHF-1) mRNA levels, which were constant during 65 nM IGF 1 treatment. MtT/S cells were shown to have both IGF-1 and insulin receptors as detected by Western blotting. There was also shown to be the suggestion of "hybrid" receptors containing different beta chains from each of these related heterotetrameric receptors. Analysis of the effects of IGF-1 and insulin on MtT/S cells showed that each reduced GH mRNA in a dose-dependent manner gave a calculated EC50 of 15.5 nM for IGF-1 and 0.6 nM for insulin, suggesting that the respective receptors for each hormone were activated. However, GH mRNA response to IGF-1 treatment was "ultrasensitive," exhibiting a switch-like effect; below 10 nM IGF-1, there was no decline in GH mRNA, but then maximal reduction occurred at IGF-1 concentrations above 20 nM. The degree of this ultrasensitive effect was calculated from the Hill equation for cooperativity, with a Hill coefficient of 4.1, greater than the classic cooperativity exhibited by hemoglobin binding to oxygen. The ultrasensitive response was specific for IGF-1, as insulin did not display this effect. These results suggest that the response evoked by the IGF-1 receptor could act as a binary molecular switch controlling GH mRNA expression in somatotrophs. PMID- 11051050 TI - MRNA expression of Phex in mice and rats: the effect of low phosphate diet. AB - Phex is the gene whose mutation is the cause of X-linked hypophosphatemia in humans and mice. The organs expressing Phex in normal animals, and their possible sensitivity to stimulation by low phosphate diets, are unknown. In this study, Phex expression was measured in 6-wk-old normal B6C3H male and female mice and in 135 g Sprague-Dawley rats fed a normal phosphate diet or a low phosphate diet with deionized water ad libitum for 7 d. The animals were then anesthetized, and a variety of organs were collected and frozen in liquid nitrogen. Phex mRNA expression was measured in each organ by reverse transcription-polymerase chain reaction (RT-PCR) with primers specific for both Phex and glyceraldehyde-3 phosphate dehydrogenase (G3PDH). Southern blots were prepared, hybridized with 32P-labeled internal oligonucleotides, and quantified with a phosphor imager. The Phex/G3PDH ratio was computed, and the data were compiled as the mean +/- SEM. In these growing animals, the highest Phexexpression levels were found in the gonads, brain, and lung. In contrast, Phex expression in calvaria and femur was markedly less. Two significant changes were found in animals that were fed a low phosphate diet. Spleen showed a significant decrease in Phex mRNA levels on low phosphate diet (60+/-10% of normal P diet, n = 12/group, p = 0.002). The pituitary gland showed a significant increase in Phex expression with low phosphate diet (851+/-127% of G3PDH) over normal P diet (569+/-78%, n = 24 - 25/group, p = 0.03). No significant change was found in femur, calvaria, or a variety of soft tissues. In summary, Phex mRNA was found in most tissues examined. Expression levels varied by two orders of magnitude from highest to lowest with more in gonads, brain, and lung and with less in bone. Increased Phex mRNA was found in the pituitary gland of animals that were fed a low phosphate diet. PMID- 11051052 TI - Pancreatic fate of 6-deoxy-6-[125I]iodo-D-glucose: in vivo experiments. AB - The fate of 6-deoxy-6-[125I]iodo-D-glucose (6-DIG), injected intravenously, was compared in control rats and animals that had received streptozotocin and were then treated with insulin or not. In the control rats, the measurement of plasma radioactivity suggested that, after an initial and rapid (up to min 10) distribution phenomenon (Kvalue: 12.2 x 10(-2) min(-1)), the clearance of the iodinated hexose occurred mainly by glomerular filtration (K value: 0.2 x 10(-2) min(-1)). Three minutes after the injection of 6-DIG, the radioactive content of muscle, liver, and pancreas, relative to the paired value in blood, was lower in untreated diabetic rats than in control animals. In the case of muscle and liver, such a difference was no longer observed when the treatment of the diabetic rats by insulin resulted in restoration of normoglycemia. In the pancreas, however, the radioactive content, whether expressed relative to the paired blood or liver value, remained significantly lower in the insulin-treated diabetic rats than in the control animals. No significant difference between control and diabetic rats, in terms of pancreatic radioactivity, was observed 10 min after the injection of 6-DIG. These findings indicate that advantage can be taken from the vastly different time course for 6-DIG uptake by pancreatic acinar and islet cells, as recently documented in vitro, to label preferentially the endocrine moiety of the pancreatic gland shortly after 6-DIG injection. PMID- 11051051 TI - Pancreatic fate of 6-deoxy-6-[125I]iodo-D-glucose: in vitro experiments. AB - The apparent distribution space of 6-deoxy-6-[125I]iodo-D-glucose, recently proposed as a tracer of D-glucose transport, was measured in rat isolated islets, acinar tissue, and pieces of pancreas. While such a space reached a steady-state value corresponding to the 3HOH volume in pancreatic islets within 5 min, it slowly increased in pieces of pancreas and, even after 60-min incubation, remained lower than the 3HOH volume. Moreover, the net uptake of 6-deoxy-6 [125I]iodo-D-glucose by pancreatic pieces was inhibited by unlabeled 6-deoxy-6 iodo-D-glucose, D-glucose, and cytochalasin B, while being less or not affected by these agents in isolated islets. A preferential labeling of the endocrine, relative to exocrine, moiety of the pancreas was documented both by comparing, after 2 min incubation, the uptake of 6-deoxy-6-[125I]iodo-D-glucose by pieces of pancreas from normal vs streptozotocin-injected rats and by comparing the radioactive content of pancreatic islets and acinar tissue obtained from normal rats injected intravenously 3 min before sacrifice with 6-deoxy-6-[125I]iodo-D glucose. It is proposed, therefore, that advantage could conceivably be taken from the vastly different time course for the uptake of selected monosaccharides by pancreatic islets vs acinar cells in the perspective of imaging of the endocrine pancreas by a non invasive method. PMID- 11051053 TI - Insulin-like growth factor (IGF)-I stimulates IGF-I and type 1 IGF receptor expression in cultured rat granulosa cells: autocrine regulation of the intrafollicular IGF-I system. AB - A growing body of information documents the existence of a complete rat intrafollicular insulin-like growth factor (IGF)-I system replete with a ligand (IGF-I), a receptor (type 1 IGF receptor) IGF binding proteins (4 and 5), and IGFBP-directed endopeptidases (4 and 5). Previous studies have established the ability of IGF-I to promote the elaboration of granulosa cell-derived IGFBP-5 and to suppress the activity of granulosa cell-derived IGFBP-5-directed endopeptidase. It was the purpose of this article to examine the effects of treatment with IGF-I on the other components of the intrafollicular IGF system, i.e., IGF-I itself and the type 1 IGF-receptor. Granulosa cells, obtained by follicular puncture from 25-d-old estrogen-primed rats were cultured in polystyrene tubes for 72 h under serum-free conditions, in the absence or presence of the indicated agents. At the conclusion of each experiment, media were discarded, and RNA was extracted and subjected to an RNase protection assay. Treatment of cultured rat granulosa cells with IGF-I resulted in a significant 1.8-fold increase in the steady-state levels of IGF-I mRNA. No effect was noted on the total cellular DNA content thereby arguing against the possibility that the relative increase in IGF-I transcripts can be ascribed to a possible treatment-induced increase in cell number in culture. The IGF-I effect was apparent (p < 0.05) at IGF-I doses as low as 1 ng/mL, minimal additional increments being noted thereafter. Treatment with insulin and des (1-3) IGF-I proved equally effective, producing 2.0- and 2.6-fold increases, respectively, thereby suggesting that the IGF-I effect may be mediated via the type 1 IGF receptor. Treatment with IGF-I also resulted in a significant (p < 0.005) increase in type 1 IGF receptor expression (2.3-fold increase), the first significant effect being noted at the 30 ng/mL dose level. Similar results obtained for insulin and des (1-3) IGF-I thereby suggest that the ability of IGF I to upregulate the expression of its own receptor is probably type 1 IGF receptor-mediated. Taken together, these findings indicate that treatment of estrogen-primed granulosa cells with IGF-I will result in upregulation of the steady-state levels of transcripts corresponding to IGF-I itself and to its type 1 IGF receptor. These observations emphasize the importance of positive autoregulatory phenomena as determinants of the intrafollicular content of IGF-I and its receptor. PMID- 11051054 TI - The effect of fetal hypoxia on adrenocortical function in the 7-day-old rat. AB - Fetal hypoxia in late gestation is a common cause of postnatal morbidity. The purpose of the present study was to evaluate adrenal function in vivo and in vitro in 7-d-old rat pups previously exposed to normoxia or hypoxia (12% O2) during the last 2-3 d of gestation. Seven-day-old rats exposed to fetal hypoxia had a small, but significant decrease in plasma aldosterone despite no decreases in plasma ACTH or renin activity. There was a small (approx 20%) but significant decrease in the aldosterone and corticosterone response to cAMP in vitro in dispersed cells from 7-d-old pups exposed to fetal hypoxia. The aldosterone, corticosterone, and cAMP response to ACTH, however, was not altered by prior fetal hypoxia. There was also no effect of fetal hypoxia on steroidogenic enzyme expression or zonal dimension in 7-d-old rats. We conclude that fetal hypoxia in late gestation results in a subtle decrease in cAMP-stimulated steroidogenesis. Fetal hypoxia appears to have minimal effects on subsequent adrenal function in the neonatal rat. PMID- 11051055 TI - Appropriate timing of glimepiride administration in patients with type 2 diabetes millitus: a study in Mediterranean countries. AB - Sulfonylureas are used to treat patients with type 2 diabetes mellitus when diet and exercise fail. Glimepiride, a new sulfonylurea, can be administered in one daily dose, thanks to its pharmacokinetic properties. We attempted to establish the optimal time of day for the administration of Glimepiride in a group of patients from the Mediterranean area by clinical trial. No relationship was found between the time of administration and fasting blood glucose values, or HbA1c, or the frequency or severity of hypoglycemic episodes. PMID- 11051056 TI - Biological effects of single and repeated swimming stress in male rats: beneficial effects of glucocorticoids. AB - We have examined the biological effects of single (45 min at 22 degrees C) and repeated swimming stress (45 min at 22 degrees C for 7 d) using male Sprague Dawley rats. Repeated swimming for a week resulted in a significant inhibition in total body weight (25%) as compared to control unstressed animals. There was significant increase in adrenal and kidney relative weight and decreases in relative thymus weight in repeated swimming-stressed animals as compared to control animals. Repeated swimming stress resulted in almost threefold increase in plasma corticosterone levels with concomitant dramatic decrease in total glucocorticoid receptor (GR) levels in liver, thymus, and heart as compared to control unstressed animals. Interestingly, single swimming stress resulted in a significant elevation in lipid peroxidation levels in the liver and heart. In contrast, there was no change in the lipid per oxidation levels in the liver and heart between chronic stressed and control unstressed animals. Finally, both single and repeated swimming-stress animals had almost 50% reduction in plasma triglyceride levels as compared to control unstressed animals. It is concluded that elevated plasma corticosterone levels by downregulating GR during repeated swimming stress exerts beneficial effects in rats by retarding the total body weight gain and lowering plasma triglyceride levels without affecting free radicals-induced oxidative stress. PMID- 11051058 TI - Assessing and managing violence risk in outpatient settings. AB - Psychologists and other mental health professionals practicing in essentially all clinical settings are called on to assess and manage clients who may pose a risk of violence to third parties. Over the past 25 years much has been learned about the relationship between violence and mental disorder, and about assessing violence risk. In this article risk factors for violence among persons with mental disorder are reviewed, clinical assessment strategies are discussed, and a model for thinking about treatment and other types of interventions designed to minimize violence risk is offered. PMID- 11051057 TI - Uptake of tritiated glibenclamide by endocrine and exocrine pancreas. AB - Tritiated glibenclamide binds to specific receptors and is internalized in pancreatic insulin-producing B-cells. We investigated, therefore, whether tritiated glibenclamide could be used to preferentially label the endocrine, as distinct from exocrine, pancreas. In isolated rat pancreatic islets, the net uptake of 3H-glibenclamide reached within 30 min of incubation a near-equilibrium value, corresponding to an apparent distribution space close to three to four times the islet volume. In pieces of pancreas exposed up to 1 h to 3H glibenclamide, however, its apparent distribution space progressively increased and, even at the min 60 of incubation, did not exceed a third of the wet weight of the pieces. Yet, no significant difference could be detected between the time course for 3H-glibenclamide uptake by pancreatic pieces from either control animals or rats injected with streptozotocin a few days before the experiments. Likewise, no significant difference in the paired ratio between the radioactive content of the pancreas and plasma could be found between the control and diabetic rats when examined 1, 5, or 24 h after the IV administration of 3H glibenclamide. These findings indicate that the sulfonylurea does not represent a suitable tool for preferential labeling of the endocrine pancreas in the perspective of its imaging by a noninvasive procedure. PMID- 11051059 TI - Assessing violence risk among youth. AB - Despite recent declines in the reported rate of juvenile violence, there appears to be increasing public and professional concern about violent behavior among children and adolescents. Media accounts of school shootings and juvenile homicides have prompted a need to develop approaches for systematically assessing violence risk. This article describes the task of assessing general violence risk among youth, and argues that a somewhat different approach is required to assess cases where an identified or identifiable young person may pose a risk to a specifically identified or identifiable target (also referred to as "targeted violence"). Key risk factors for violent behavior among children and adolescents are identified, fundamental principles for conducting an assessment of violence potential in clinical and juvenile justice contexts are outlined, and an approach to assessment when an identified person engages in some communication or behavior of concern that brings him or her to official attention is briefly described. PMID- 11051060 TI - Risk for domestic violence: factors associated with perpetration and victimization. AB - The extent and potential dangerousness of the problem of domestic violence warrants systematic screening and assessment in all mental health settings. Few empirical studies have approached the question of domestic violence with the aim of identifying risk markers, making it impossible to identify a particular characteristic or set of characteristics that can be used to identify individuals at risk for perpetrating or becoming the victims of domestic violence. However, there are a number of factors that have been identified as correlates of domestic violence that may eventually prove useful for identifying individuals at risk, but the extant literature does not provide the empirical support at this time. Because many of these correlates may be brought to the attention of mental health and medical professionals (e.g., depression, substance use/abuse, physical injuries) and given the absence of established risk factors for domestic violence, there is a need for clinicians to systematically assess for violence among all of their patients. By identifying factors that might help clinicians realize that many of their patients are at risk for domestic violence, we hope to encourage them to attend to this potentially dangerous problem. Ongoing assessment in the context of knowledge regarding correlates of domestic violence can provide important information for evaluating risk of a particular violent incident. In addition, we outline strategies for assessing violence and violence risk in both perpetrators and victims in order to assist clinicians in approaching this difficult topic in a clinical setting. A careful assessment of the potential for violence within clients' ongoing relationships is necessary for clinicians to provide appropriate clinical care. PMID- 11051061 TI - Emergency evaluation and intervention with female victims of rape and other violence. AB - Given the high prevalence of crime within the general population and the increased rates of victimization among those seeking medical care, professionals who work in emergency departments, primary care medical facilities, or mental health settings need to be prepared to address physical and psychological problems related to sexual and physical assault. In this paper, interpersonal violence prevalence studies are reviewed in terms of study design and findings for sexual assault and physical assault. Common injuries following both forms of assault are documented, followed by a review of long-term medical outcomes. In addition to a review of physical health outcomes, primary psychological effects of violence are also reviewed. Strategies with which to screen for interpersonal violence in the medical setting are offered, and issues related to mandatory reporting are summarized. Interventions for assault victims that can be implemented in the medical setting are outlined, and a new hospital-based treatment for victims of rape is described. PMID- 11051062 TI - Risk management and life-threatening patient behaviors. AB - This article reviews the legal and ethical issues that arise in the treatment of patients who may pose a threat to harm themselves or others. The recent practice of developing empirically validated treatments has not yet been applied to the diagnosis and treatment of patients who present an imminent danger of harm to themselves or others. Instead, psychologists must follow standards that come primarily from court cases or statutes. Although these standards have some degree of commonality across North America, psychologists are encouraged to be informed about local laws and court cases. PMID- 11051063 TI - The stress of patient emergencies for the clinician: incidence, impact, and means of coping. AB - This article presents evidence from the literature on the incidence and impact of behavioral emergencies on clinicians as well as suggestions for improved education and support for work in this area. Behavioral emergencies are conceptualized as including imminent life-threatening behaviors such as patient suicidal behavior, patient violence, and instances in which patients become the victims of interpersonal violence. Suggestions are offered for how clinicians can understand and cope with their own reactions during and after such patient emergencies. Additionally, data on deficits in the education and training of psychologists are presented along with suggestions for how programs and clinical sites can improve their training in emergency and crisis work. PMID- 11051065 TI - MACI personality scale profiles of depressed adolescent suicide attempters: a pilot study. Million Adolescent Clinical Inventory. AB - Employing the Millon Adolescent Clinical Inventory (MACI), we examined differences in the maladaptive personality style profiles of clinically referred, depressed adolescents presenting with (n = 26) and without (n = 23) a history of previous suicide attempts. Relative to the comparison group, adolescent attempters experienced more severe overall levels of personality dysfunction. At the trait level, attempters obtained higher scores on the forceful and borderline tendency scales and lower scores on the submissive and conforming scales of the MACI, reflecting negative mood regulation deficits (e.g., anger control problems) and persistently high levels of aggressive impulsivity. These preliminary findings suggest that MACI personality scales may be useful in discriminating adolescents with and without previous suicidal behavior, especially among depressed outpatient samples. PMID- 11051064 TI - Trauma-related psychophysiological reactivity in women exposed to war-zone stress. AB - Women are at particular risk for Posttraumatic Stress Disorder (PTSD), but surprisingly little is known about their objective manifestations of the disorder's hallmark symptoms. Although research suggests that people with PTSD exhibit physiological reactivity to the presentation of trauma-related cues, the majority of studies to date have focused on men. We assessed the physiological reactions of three groups of trauma-exposed female Vietnam veterans (those with current PTSD, lifetime PTSD, or no PTSD) to war-related stimuli. Responses of women with current PTSD differed significantly from those without PTSD on skin conductance and systolic blood pressure, and mean levels of reactivity for women with lifetime PTSD fell between the other two groups. Although symptom severity was correlated with physiologic reactivity overall, results suggested differential relationships at the symptom cluster level. Study results replicate earlier findings with men and extend knowledge of autonomic reactivity to an important group of female survivors. PMID- 11051066 TI - Risk-based design of aseptic processing of heterogeneous food products. AB - A risk assessment was performed to incorporate uncertainty in food processing conditions to develop a risk-based sterilization process design. The focus of this analysis was uncertainty associated with heterogeneous food products. Quartered button mushrooms were the chosen food product because it represents the most typical type. A model for sterilization of spherical particles was utilized, and each parameter's uncertainty was characterized for use under Monte Carlo simulation. Various particle distributions and fluid types were compared. The output of the model was the required sterilization time to achieve the target sterilization conditions with 95% probability. This value was then used to determine the mean fluid velocity for a given tube length. Finally, the output from the model was analyzed to determine the confidence in output based on uncertainty in the input parameters. The model was more sensitive to variation in particle size distribution than fluid type for power-law fluids. The 90% confidence interval included a holding time range of 1 min. With a 95% confidence level that only 8% of the data will be below the target sterilization conditions, a maximum of 9% of the data were expected to achieve double the target level. The results of such an analysis would be useful for management decisions concerning the design of aseptic food processing operations. PMID- 11051067 TI - Determinants of priority for risk reduction: the role of worry. AB - One hundred twenty-two members (experts) of the Society for Risk Analysis completed a mailed questionnaire and 150 nonexperts completed a similar questionnaire on the World Wide Web. Questions asked included those about priorities on personal and government action for risk reduction, badness of the risk, number of people affected, worry, and probabilities for self and others. Individual differences in mean desire for action were largely explained in terms of worry. Worry, in turn, was largely affected by probability judgments, which were lower for experts than for nonexperts. Differences across risks in the desire for action, within each subject, were also determined largely by worry and probability. Belief in expert knowledge about the risk increased worry and the priority for risk reduction. A second study involving 91 nonexperts (42 interviewed and 49 on the Web) replicated the main findings for nonexperts from the first study. Interviews also probed the determinants of worry, attitudes toward government versus personal control, and protective behaviors. PMID- 11051068 TI - Assessing overall risk in reproductive experiments. AB - Toxicologists are often interested in assessing the joint effect of an exposure on multiple reproductive endpoints, including early loss, fetal death, and malformation. Exposures that occur prior to mating or extremely early in development can adversely affect the number of implantation sites or fetuses that form within each dam and may even prevent pregnancy. A simple approach for assessing overall adverse effects in such studies is to consider fetuses or implants that fail to develop due to exposure as missing data. The missing data can be imputed, and standard methods for the analysis of quantal response data can then be used for quantitative risk assessment or testing. In this article, a new bias-corrected imputation procedure is proposed and evaluated. The procedure is straightforward to implement in standard statistical packages and has excellent operating characteristics when used in combination with a marginal model fit with generalized estimating equations. The methods are applied to data from a reproductive toxicity study of Nitrofurazone conducted by the National Toxicology Program. PMID- 11051069 TI - A systematic uncertainty analysis of an evaluative fate and exposure model. AB - Multimedia fate and exposure models are widely used to regulate the release of toxic chemicals, to set cleanup standards for contaminated sites, and to evaluate emissions in life-cycle assessment. CalTOX, one of these models, is used to calculate the potential dose, an outcome that is combined with the toxicity of the chemical to determine the Human Toxicity Potential (HTP), used to aggregate and compare emissions. The comprehensive assessment of the uncertainty in the potential dose calculation in this article serves to provide the information necessary to evaluate the reliability of decisions based on the HTP A framework for uncertainty analysis in multimedia risk assessment is proposed and evaluated with four types of uncertainty. Parameter uncertainty is assessed through Monte Carlo analysis. The variability in landscape parameters is assessed through a comparison of potential dose calculations for different regions in the United States. Decision rule uncertainty is explored through a comparison of the HTP values under open and closed system boundaries. Model uncertainty is evaluated through two case studies, one using alternative formulations for calculating the plant concentration and the other testing the steady state assumption for wet deposition. This investigation shows that steady state conditions for the removal of chemicals from the atmosphere are not appropriate and result in an underestimate of the potential dose for 25% of the 336 chemicals evaluated. PMID- 11051070 TI - Multiattribute risk analysis in nuclear emergency management. AB - Radiation protection authorities have seen a potential for applying multiattribute risk analysis in nuclear emergency management and planning to deal with conflicting objectives, different parties involved, and uncertainties. This type of approach is expected to help in the following areas: to ensure that all relevant attributes are considered in decision making; to enhance communication between the concerned parties, including the public; and to provide a method for explicitly including risk analysis in the process. A multiattribute utility theory analysis was used to select a strategy for protecting the population after a simulated nuclear accident. The value-focused approach and the use of a neutral facilitator were identified as being useful. PMID- 11051071 TI - A modeling framework for exposing risks in complex systems. AB - This article introduces and develops a modeling framework for exposing risks in the form of human errors and adverse consequences in high-risk systems. The modeling framework is based on two components: a two-dimensional theory of accidents in systems developed by Perrow in 1984, and the concept of multiple system perspectives. The theory of accidents differentiates systems on the basis of two sets of attributes. One set characterizes the degree to which systems are interactively complex; the other emphasizes the extent to which systems are tightly coupled. The concept of multiple perspectives provides alternative descriptions of the entire system that serve to enhance insight into system processes. The usefulness of these two model components derives from a modeling framework that cross-links them, enabling a variety of work contexts to be exposed and understood that would otherwise be very difficult or impossible to identify. The model components and the modeling framework are illustrated in the case of a large and comprehensive trauma care system. In addition to its general utility in the area of risk analysis, this methodology may be valuable in applications of current methods of human and system reliability analysis in complex and continually evolving high-risk systems. PMID- 11051072 TI - Emergency planning for hazardous industrial areas: a Brazilian case study. AB - One of the characteristics of modern industrial development is the emergence of a new typology of accidents whose effects can be spread, in space as well as in time, well beyond the borders of the installations where they occur, sometimes impacting the local population and the environment in a catastrophic fashion. This is the result of a number of factors that have changed the risk profile of modern industrial activities. For a number of reasons, the developing countries have proved to be more vulnerable to industrial disasters. Three of the most catastrophic industrial accidents--Bhopal, San Juan de Ixhuatepec, and Cubatao- occurred in developing countries, claiming thousands of lives. During the 1970s and 1980s the higher degree of public visibility of industrial hazards as a result of serious accidents, led to the creation, especially in the more industrialized countries, of regulations for greater control over industrial activities, either by means of new laws or by updating existing legislation. Some of these regulations were designed to improve the response to accidents with potential impacts outside the industrial sites. This article attempts to describe the current status and identify the shortcomings of off-site emergency planning for hazardous industrial areas in Brazil. The most important problems are the lack of specific legislation and the absence of awareness and active participation of public authorities. The experience of an off-site emergency planning process for a Brazilian industrial area is presented. This experience illustrates how difficult it is to prepare and implement emergency planning processes in an industrializing country. PMID- 11051073 TI - An integrated risk model of a drinking-water-borne cryptosporidiosis outbreak. AB - A dynamic risk model is developed to track the occurrence and evolution of a drinking-water-borne cryptosporidiosis outbreak. The model characterizes and integrates the various environmental, medical, institutional, and behavioral factors that determine outbreak development and outcome. These include contaminant delivery and detection, water treatment efficiency, the timing of interventions, and the choices that people make when confronted with a known or suspected risk. The model is used to evaluate the efficacy of alternative strategies for improving risk management during an outbreak, and to identify priorities for improvements in the public health system. Modeling results indicate that the greatest opportunity for curtailing a large outbreak is realized by minimizing delays in identifying and correcting a drinking-water problem. If these delays cannot be reduced, then the effectiveness of risk communication in preemptively reaching and persuading target populations to avoid exposure becomes important. PMID- 11051074 TI - The Beta Poisson dose-response model is not a single-hit model. AB - The choice of a dose-response model is decisive for the outcome of quantitative risk assessment. Single-hit models have played a prominent role in dose-response assessment for pathogenic microorganisms, since their introduction. Hit theory models are based on a few simple concepts that are attractive for their clarity and plausibility. These models, in particular the Beta Poisson model, are used for extrapolation of experimental dose-response data to low doses, as are often present in drinking water or food products. Unfortunately, the Beta Poisson model, as it is used throughout the microbial risk literature, is an approximation whose validity is not widely known. The exact functional relation is numerically complex, especially for use in optimization or uncertainty analysis. Here it is shown that although the discrepancy between the Beta Poisson formula and the exact function is not very large for many data sets, the differences are greatest at low doses--the region of interest for many risk applications. Errors may become very large, however, in the results of uncertainty analysis, or when the data contain little low-dose information. One striking property of the exact single-hit model is that it has a maximum risk curve, limiting the upper confidence level of the dose-response relation. This is due to the fact that the risk cannot exceed the probability of exposure, a property that is not retained in the Beta Poisson approximation. This maximum possible response curve is important for uncertainty analysis, and for risk assessment of pathogens with unknown properties. PMID- 11051075 TI - Child passenger safety: decisions about seating location, airbag exposure, and restraint use. AB - The installation of passenger-side airbags in new vehicles complicates efforts to maximize child safety in motor vehicle crashes. It has been recommended by both public and private organizations that children sit in the rear seat with proper restraint to achieve maximum safety. Drivers now need to decide whether a child should be restrained, where the child should be seated (front versus rear), and whether the child should be seated in front of a passenger-side airbag. This research was undertaken to determine which choice minimizes the risk of fatality to children. Using data from the U.S. Fatality Analysis Reporting System for calendar years 1989 to 1998, fatal vehicle crashes with child passengers younger than 13 years were analyzed. The effectiveness of passenger-side airbags and rear seating for children, by age category and restraint use, was estimated using the double-pair comparison method. For each of four age categories, the fatality risk of each possible combination of restraint use, seating location, and airbag presence was also estimated using logistic regression. Passenger airbags were associated with an increase in child fatality risk of 31% for restrained children, and 84% for unrestrained children. Passenger airbags did appear to offer protection to restrained 9- to 12-year-old children. Restraint use and rear seating were associated with statistically significant reductions in the odds of a child dying in a crash. In order to minimize child fatality risk, parents should seat children in the rear of the vehicle while using the proper child restraint system, especially in vehicles with passenger airbags. These findings support current public education efforts in the United States. PMID- 11051077 TI - Modelmaker 4.0. PMID- 11051076 TI - Use of quality-adjusted life year weights with dose-response models for public health decisions: a case study of the risks and benefits of fish consumption. AB - Risks associated with toxicants in food are often controlled by exposure reduction. When exposure recommendations are developed for foods with both harmful and beneficial qualities, however, they must balance the associated risks and benefits to maximize public health. Although quantitative methods are commonly used to evaluate health risks, such methods have not been generally applied to evaluating the health benefits associated with environmental exposures. A quantitative method for risk-benefit analysis is presented that allows for consideration of diverse health endpoints that differ in their impact (i.e., duration and severity) using dose-response modeling weighted by quality adjusted life years saved. To demonstrate the usefulness of this method, the risks and benefits of fish consumption are evaluated using a single health risk and health benefit endpoint. Benefits are defined as the decrease in myocardial infarction mortality resulting from fish consumption, and risks are defined as the increase in neurodevelopmental delay (i.e., talking) resulting from prenatal methylmercury exposure. Fish consumption rates are based on information from Washington State. Using the proposed framework, the net health impact of eating fish is estimated in either a whole population or a population consisting of women of childbearing age and their children. It is demonstrated that across a range of fish methylmercury concentrations (0-1 ppm) and intake levels (0-25 g/day), individuals would have to weight the neurodevelopmental effects 6 times more (in the whole population) or 250 times less (among women of child-bearing age and their children) than the myocardial infarction benefits in order to be ambivalent about whether or not to consume fish. These methods can be generalized to evaluate the merits of other public health and risk management programs that involve trade-offs between risks and benefits. PMID- 11051078 TI - Sustained oscillations of transmembrane Ca2+ fluxes in mitochondria and their possible biological significance. AB - Sustained oscillations of transmembrane fluxes of Ca2+ and other ions in isolated mitochondria are described. The data are presented that the major cause of the oscillations is the Ca2+-induced Ca2+ efflux from the mitochondrial matrix and spontaneous opening/closing of the permeability transition pore in the inner mitochondrial membrane. Conditions favourable for the generation of oscillations are considered. The role of phospholipid peroxidation and hydrolysis in the generation of [Ca2+] oscillations is emphasized. Literature data concerning [Ca2+] changes in the mitochondrial matrix in intact cells and the data on the participation of mitochondria in intracellular Ca2+ oscillation and in the Ca2+ wave propagation are reviewed. The hypothesis that mitochondria are able to generate [Ca2+] oscillations in intact cells is put forward. It is assumed that Ca2+ oscillations can protect mitochondria of resting cells from osmotic shock and oxidative stress. PMID- 11051079 TI - Dynamics of structural and functional association of nucleolar chromosomes in cells of the hexaploid wheat Triticum aestivum L. at different stages of the cell cycle and during genome polyploidization. AB - Quantitative analysis of interphase association of the nucleolar chromosomes at different stages of the cell cycle and during genome polyploidization was carried out. Cells of various tissues of hexaploid wheat Triticum aestivum L. (Moskovskaya-35) were used, including diploid root meristematic cells, endopolyploid root cells, triploid endosperm cells and antipodal cells with polytene chromosomes. Interphase nucleoli impregnated with silver or stained with autoimmune antibodies to 53 kDa nucleolar protein served as markers of the nucleolar chromosome association. The following data were obtained: (1) silver staining revealed two pairs of homologous chromosomes 1B and 6B with active nucleolus-organizing regions in the root meristematic cells; (2) maximal number of nucleoli in diploid meristematic cells reaches four, which corresponds to the number of chromosomes with active organizers; (3) analysis of cells at different stages of the cell cycle has shown that the tendency to the nucleoli association is observed as soon as cells pass individual stages of the cycle; (4) after DNA and chromosome reduplication, the nucleolus-organizing regions in sister chromatids function as a common structure-functional complex; (5) in endopolyploid root cells and antipodal cells with polytene chromosomes, the number of nucleoli does not correlate with ploidy level, and an additional nucleolus revealed in some cells is the result of activation of the latent organizer in one of the nucleolar chromosomes; (6) in the triploid endosperm nucleologenesis, the stage of prenucleolar bodies is missing. Our data suggest that "fusion" of nucleoli and reduction of their number due to the "satellite" association of the nucleolar chromosomes are two independent processes regulated by different mechanisms. PMID- 11051080 TI - Effect of dipyridamole on the recombination kinetics between photooxidized bacteriochlorophyll and photoreduced primary quinone in reaction centres of purple bacteria. AB - The action of dipyridamole (DIP) on dark recombination between the photooxidized special pair bacteriochlorophyll BChl2+ and reduced primary quinone acceptor Q(A) in the reaction centres (RCs) of the bacteria Rhodobacter sphaeroides was studied in the presence of different detergents (LDAO, Triton X-100, sodium cholate, sodium dodecyl sulfate). DIP accelerated this reaction approximately 4-5 fold. In RCs with the extracted H-subunit, the effect of DIP was observed at lower concentrations. The possibility of modification of the RC structure-dynamic state by DIP (including changes in RC hydrogen bonds) is proposed. The modification obviously disturbs the processes of the long-life electrostatic stabilization of Q(A)-. PMID- 11051081 TI - Effect of the combined action of flavonoids, ascorbate and alpha-tocopherol on peroxidation of phospholipid liposomes induced by Fe2+ ions. AB - The effect of alpha-tocopherol, ascorbate, rutin and dihydroquercetin on chemiluminescence (CL) accompanying the Fe2+-induced peroxidation of unsaturated fatty acids in phospholipid liposomes has been investigated. The amplitude of CL decreased and the latent period increased in the presence of alpha-tocopherol, rutin and dihydroquercetin which is typical of peroxide radical traps. Ascorbate also reduced the CL amplitude but only at small concentrations up to about 4 microM. A further increase of ascorbate concentration had a negligible effect on the amplitude. At the same time, the latent period in CL development increased with the growth of ascorbate concentration, apparently, as a result of recycling of divalent iron oxidized in the course of lipid peroxidation. The effects of rutin and dihydroquercetin on the liposomal CL in the presence of alpha tocopherol and ascorbate in all experiments were almost the same as when these compounds were added individually. The antioxidant effects were merely summed up without any mutual enhancement or inhibition of each other's action. PMID- 11051082 TI - Interphase microtubules in cultured cells: long or short? AB - Presently, the question about the length of microtubules in the interphase cell became actual, since the parameters of dynamic instability of the plus end measured in vivo do not allow one to explain the rapid turnover of the long microtubule system. The problem may be solved if one of the following suppositions is assumed: either microtubules undergo rapid depolymerization from the minus end or they are on the average much shorter than it is usually considered. To check the last hypothesis, we have reconstructed microtubules using stereophotography of electron microscopic sections. Microtubules around the cell center in cultures of epithelial cells (kidney of pig embryo (PK) and bovine trachea (FBT)) and fibroblasts (MEF, primary mouse embryo fibroblasts, and L cells), as well as at the periphery of PK cells were studied. All in all, no less than 200 microtubules were found near the centrosome in each cell culture. From 2.5 to 8% microtubules were beyond the studied volume (4.0 x 5.5 x 1.5 microm). Most of microtubules in all studied cell lines were up to 1 microm and about 1/3 of them were 0.2-0.4 microm long. The mean length of microtubules surrounding the centrosome in different cell lines differed insignificantly and equalled 0.4-0.8 microm. In this case, the microtubules attached to the centrosome were on the average slightly shorter than the free ones. Thus, almost all microtubules around the centrosome are short, and the majority of those attached to it do not reach the cell periphery. A similar reconstruction of a part of the PK cell cytoplasm (10 x 35 microm) has shown that at the periphery, the mean length of microtubules is about 1.6 microm and most of them are 0.5 to 1.5 microm long. Thus, our data confirm the recent hypothesis of Vorobjev et al. (I. A. Vorobjev, T. M. Svitkina, and G. G. Borisy, J. Cell Sci. 110:2635-2645 (1997)) that most of microtubules in the cells are not connected with the centrosomes. PMID- 11051083 TI - The role of actin cytoskeleton in the generation of surface oscillations of red blood cell ghosts. AB - We have studied the effects of three compounds on surface oscillations of human red blood cell ghosts: the P-ATPase inhibitor, suramin; the fluorescent dye of a similar structure, 1,8-anilinonaphthalene sulfonate (ANS); and subfragment 1 of skeletal muscle myosin (S1). It has been found that suramin (10 microM), ANS (100 microM) and S1 (2 mg/ml) suppress the surface oscillations reversibly. The shape of the ghosts remains unchanged. We have also found that suramin and ANS inhibit the ghosts' non-transport (presumably, F-actin-associated) ATPase. The results of the present study suggest the important role of actin ATPase in the generation of cell surface oscillations. The effect of S1, the protein which increases the torsional, but not the bending, rigidity of F-actin upon binding to filaments, favours the possibility that just the torsional dynamics of actin protofilaments leads to the observed oscillations of the ghosts' surface. PMID- 11051084 TI - Oligomeric state of the muscle glycogen phosphorylase b in the system of reversed micelles of aerosol OT in octane. AB - The oligomeric state and formation of supramolecular structures of glycogen phosphorylase b from rabbit skeletal muscle was studied in the system of aerosol OT (AOT) reversed micelles in octane. The sedimentation experiments have shown that the enzyme oligomeric state depends on the degree of micelle hydration. The enzyme monomer, dimer, trimer, tetramer, hexamer, and octamer were observed, depending on the degree of hydration. PMID- 11051085 TI - A fast method for the quantitative estimation of the distribution of hydrophobic and hydrophilic segments in alpha-helices of membrane proteins. AB - The work presents a fast quantitative approach for estimating the orientations of hydrophilic and hydrophobic regions in the helical wheels of membrane-spanning alpha-helices of transmembrane proteins. The common hydropathy analysis provides an estimate of the integral hydrophobicity in a moving window which scans an amino acid sequence. The new parameter, orientation hydrophobicity, is based on the estimate of hydrophobicity of the angular segment that scans the helical wheel of a given amino acid sequence. The corresponding procedure involves the treatment of transmembrane helices as cylinders with equal surface elements for each amino acid residue. The orientation hydrophobicity, P(phi), phi = 0-360 degrees, of a helical cylinder is given as a sum of hydrophobicities of individual amino acids which are taken as the S-shaped functions of the angle between the centre of amino acid surface element and the centre of the segment. Non-zero contribution to P(phi) comes only from the amino acids belonging to the angular segment for a given angle phi. The size of the angular segment is related to the size of the channel pore. The amplitudes of amino acid S-functions are calibrated in the way that their maximum values (reached when the amino acid is completely exposed into the pore) are equal to the corresponding hydropathy index in the selected scale (here taken as Goldman-Engelman-Steitz hydropathy scale). The given procedure is applied in the studies of three ionic channels with well characterized three-dimensional structures where the channel pore is formed by a bundle of alpha-helices: cholera toxin B, nicotinic acetylcholine homopentameric alpha7 receptor, and phospholamban. The estimated maximum of hydrophilic properties at the helical wheels are in a good agreement with the spatial orientations of alpha-helices in the corresponding channel pores. PMID- 11051086 TI - Reduction of Ca2+-transporting systems in memory T cells. AB - Antigen-specific B and T lymphocytes make up the material grounds of immune memory, their main functional distinction from the so-called "naive" cells is due to the rapid and enhanced response to the antigen-pathogen. An essential distinction between the memory and naive T cells is different sensitivity of these two subpopulations of T lymphocytes to Ca2+-ionophores. Comparative analysis of Ca2+ responses of the immune memory T lymphocytes and naive T cells of mouse CBA/J line to the addition of Ca2+-mobilizing agents concanavalin A, thapsigargin, and ionomycin was carried out. These compounds in concentrations increasing [Ca2+]i in naive cells had no effect on [Ca2+]i in memory cells. Thus, the Ca2+ entrance into memory cells was not activated by exhaustion of intracellular resources. Estimation of intracellular resources of Ca2+, mobilized by ionomycin and thapsigargin in Ca2+ free medium has shown the absence in memory T cells of the intracellular Ca2+ pool, which may be one of factors of their resistance to ionophores. Reduction of the system of Ca2+ influx into memory T cells was shown using the SH-reagent thimerosal. Memory T cells appear to be resistant to "Ca2+ -paradox." Their incubation with 0.5 mM EDTA in the presence or absence of Ca2+ -mobilizing compounds followed by addition of 2 mM CaCl2 did not result in induction of Ca2+ influx into these cells. PMID- 11051087 TI - The regulation of L-type Ca2+ currents in rat cardiac myocytes. AB - The regulation of L-type Ca2+ current in isolated rat cardiac cells was studied using the perforated patch-clamp technique. A dual effect of the cAMP-dependent phosphorylation activator, isoproterenol, at different holding potentials (V(h)) was shown. The currents increased at V(h) = -50 mV and decreased at V(h) = -30 mV. A dihydropyridine agonist, BAY K 8644, and isoproterenol had an additive effect on the activation of Ca2+ channels at holding potentials close to the resting potential. The additivity was disturbed at more positive V(h). The activating effect of BAY K 8644 did not virtually change in the presence of a protein kinase blocker, H8, and a phosphatase activator, acetylcholine. The results were interpreted within the framework of a two-site phosphorylation model with two independent pathways of Ca2+ current regulation. PMID- 11051088 TI - Critical analysis of the impedance method for the evaluation of permittivity and conductivity of the plasma membrane. AB - We report a critical analysis of a typical method of dielectric spectroscopy consisting in impedance measurements as a function of frequency. Experimental data were obtained by measuring impedance on human erythrocyte suspensions. Since these cells do not have a nucleus they represent an ideal material for the application of the well established single shell model. This allows the evaluation of permittivity and conductivity of the plasma membrane. We discuss the influence on the reliability of results of parameters such as fractional volume, average dimensions and membrane thickness of cells. PMID- 11051089 TI - Distribution of key enzymes of purine biosynthesis in fractions of mammalian retina. PMID- 11051090 TI - Low-barrier hydrogen bonds and enzymatic catalysis. AB - Short, strong (low barrier) hydrogen bonds occur when the pK values of the atoms sharing the proton are similar. The overall distance is 2.5 A or less, the deuterium fractionation factor is less than 0.5, the proton NMR chemical shift can approach 20 ppm, and deuterium or tritium substitution causes an up-field change in the chemical shift. Such bonds can have deltaH values of 25 kcal/mol in the gas phase, and at least half that in water or other high-dielectric medium. The strength of the hydrogen bond in an active site drops by approximately 1 kcal/mol for each pH unit mismatch in pKs. When a weak hydrogen bond in the initial enzyme-substrate complex is converted into a low-barrier one by alteration of the pK of the substrate or catalytic group so that the pKs match, the increase in hydrogen bond strength can be used to help catalyze the reaction. A well-established example of this is the reaction catalyzed by serine proteases. The pK of neutral histidine is 14, while that of aspartate is approximately 6. Proton transfer from serine to permit attack on bound substrate produces protonated histidine, with a pK now matching that of aspartate. Studies with trifluoromethyl ketone inhibitors that form tetrahedral adducts show up to five orders of magnitude in binding strength as the result of formation of a low barrier hydrogen bond between aspartate and histidine. Other enzymes whose mechanisms appear to involve low-barrier hydrogen bonds include liver alcohol dehydrogenase, steroid isomerase, triose-P isomerase, aconitase, citrate synthase, and zinc proteases. It is likely that low-barrier hydrogen bonds form at the transition state of any reaction involving general-acid or general-base catalysis, as at that point the pKs of the catalytic group and reactant will be equal. PMID- 11051091 TI - Radical mechanisms in adenosylmethionine- and adenosylcobalamin-dependent enzymatic reactions. AB - A class of enzymatic reactions of S-adenosylmethionine (AdoMet) has recently been recognized, in which AdoMet plays a novel role by initiating free radical formation through the intermediate formation of 5'-deoxyadenosine-5'-yl, the 5' deoxyadenosyl radical. The reactions are in this way related to adenosylcobalamin dependent processes, which also depend on the formation of the 5'-deoxyadenosyl radical as an intermediate. The mechanisms by which the 5'-deoxyadenosyl radical is generated by the AdoMet- and adenosylcobalamin-dependent enzymes are very different. However, the functions of the 5'-deoxyadenosyl radical are similar in that in all cases it abstracts hydrogen from a substrate to form 5' deoxyadenosine and a substrate-derived free radical. In this paper, the role of the 5'-deoxyadenosyl radical in the reaction of the adenosylcobalamin-dependent reactions will be compared with its role in the AdoMet-dependent reaction of lysine 2,3-aminomutase. The mechanism by which AdoMet is cleaved to the 5' deoxyadenosyl radical at enzymatic sites will also be discussed. PMID- 11051092 TI - Oxidation-reduction properties of two engineered redox-sensitive mutant Escherichia coli malate dehydrogenases. AB - Redox potentials for two inactivating intrasubunit disulfides that link helix-5 and helix-9 in mutant Escherichia coli malate dehydrogenases have been determined. The Em is -285 mV when cysteines are at positions 121 and 305 and 295 mV when the cysteines are at positions 122 and 305. Oxidation to the disulfide affects kcat but not Km values. In the single V121C and N122C mutants, the Cys in helix-5 affects the Km for oxalacetate. The pH optimum in the direction of malate formation is affected by the redox state of the enzyme. Clearly, a disulfide bond can and does form between Cys residues substituted into positions 121 or 122 in the nucleotide binding domain and 305 in the carbon substrate binding domain of this NAD-dependent malate dehydrogenase. Apparently, crosslinking the domains interferes with catalysis. PMID- 11051093 TI - A picomolar inhibitor of resistant strains of human immunodeficiency virus protease identified by a combinatorial approach. AB - In order to identify inhibitors of various drug-resistant forms of the human immunodeficiency virus protease (HIV PR), we have designed and synthesized pseudopeptide libraries with a general structure Z-mimetic-Aa1-Aa2-NH2. Five different chemistries for peptide bond replacement have been employed and the resulting five individual sublibraries tested with the HIV PR and its drug resistant mutants. Each mutant contains amino acid substitutions that have previously been shown to be associated with resistance to protease inhibitors, including Ritonavir, Indinavir, and Saquinavir. We have mapped the subsite preferences of resistant HIV PR species with the aim of selecting a pluripotent pharmaceutical lead. All of the enzyme species in this study manifest clear preference for an L-Glu residue in the P2' position. Slight, but significant, differences in P3' subsite specificity among individual resistant PR species have been documented. We have identified three compounds, combining the most favorable features of the inhibitor array, that exhibit low-nanomolar or picomolar Ki values for all three mutant PR species tested. PMID- 11051094 TI - Affinity purification and characterization of a cutinase from the fungal plant pathogen Monilinia fructicola (Wint.) honey. AB - Trifluoromethyl ketones (TFK) are potent inhibitors of a variety of serine hydrolases. The TFK inhibitor, 3-(4-mercaptobutylthio)-1,1,1-trifluoro-2 propanone (MBTFP), was found to competitively inhibit cutinase activity (I50 = 9.4 x 10(-3)) from the fungal plant pathogen Monilinia fructicola and to serve as an effective affinity ligand for the purification of cutinases from culture filtrates. The TFK inhibitors, 3-n-octylthio-1,1,1-trifluoro-2-propanone (OTFP) and 3-n-pentylthio-1,1,1-trifluoro-2-propanone (PTFP), also inhibited cutinase activity with I50 values of 1.6 x 10(-6) and 2.3 x 10(-4) M, respectively. Buffer containing OTFP was the strongest eluant for cutinases of M. fructicola and provided the best purification factor and yield, although buffers containing OTFP, detergent, and salt were found to be effective for eluting cutinases bound to MBTFP-Sepharose. Buffer containing 0.5% Triton X-100 also selectively eluted cutinases from the affinity column. Two-dimensional electrophoretic analysis by SDS-PAGE and isoelectric focusing of the affinity-purified cutinase fraction indicated activity associated with proteins of pI 8.2 and molecular masses of approximately 18.6 and 20.8 kDa. These proteins hydrolyzed [3H]cutin and artificial substrates such as p-nitrophenylbutyrate and related esters, typical of other cutinases, but differ from previously characterized cutinases in molecular mass. The two low-molecular-weight proteins resolved by 2-D gel electrophoresis were subjected to in-gel digestion with Lys-C and the resulting peptide fragments were separated by Microbore-HPLC. The amino acid sequences of several internal peptide fragments had high homology with cutinase sequences from other fungi, particularly the plant pathogen Botrytis cinerea. Our study illustrates the potential of TFK ligands for the affinity purification of cutinases and indicates that the cutinases from M. fructicola have novel features warranting further study. PMID- 11051095 TI - Lipid-protein interactions in rat renal subcellular membranes: a biophysical and biochemical study. AB - The phase behavior of plasma membrane (PM), endoplasmic reticulum (ER), and nuclear membranes (NM) isolated from adult rat papillary cells was studied using the molecular probe Laurdan. The steady-state fluorescence data analysis was correlated with the lipid composition obtained by biochemical assays. The comparison between intact membranes and protein-free reconstituted vesicles using the whole lipid extract shows the essential role of proteins on the temperature response of natural membranes. The phospholipid (PL) and cholesterol (Cho) content was measured in the three membrane fractions, the PL/Cho molar ratio being between 1.5 and 1.9. However, Laurdan's parameters in NM show a fluid phase state pattern even at low temperature (5 degrees C), with a restricted dipole relaxation in comparison with that displayed in liquid crystalline phase state lipid model membranes. PM and ER are in a gel-like state at temperatures below 20 degrees C, showing increasing dipole relaxation with temperature. The curved fits obtained are characteristic of cholesterol-enriched membranes. The distinctive phase behavior of nuclear membranes vanishes when proteins are extracted. However, relaxation is still faster in this fraction, which correlates with the native lipid composition. NM has the lowest percentage of phosphatidylinositol and sphingomyelin-the latter being a highly saturated phospholipid- and the highest percentage of phosphatidylcholine and phosphatidylethanolamine (PE), nuclear PE being enriched in arachidonic acid. All these changes agree with the higher fluidity of NM compared with ER or PM in the conditions assayed. PMID- 11051096 TI - Oxidants depress the synthesis of phosphatidylinositol 4,5-bisphosphate in heart sarcolemma. AB - Phosphatidylinositol 4,5-bisphosphate (PtdIns 4,5-P2) is the substrate for phosphoinositide-phospholipase C (PLC) and is required for the function of several cardiac cell plasma membrane (sarcolemma, SL) proteins. PtdIns 4,5-P2 is synthesized in the SL membrane by coordinated and successive actions of PtdIns 4 kinase and PtdIns 4-phosphate 5-kinase. These kinases and the generation of PtdIns 4,5-P2 may be a factor in the cardiac dysfunction during pathophysiological conditions of oxidative stress. Therefore, we examined the effects of different reactive oxygen species (ROS) on the kinases' activities and subsequent generation of PtdIns 4,5-P2. Exposure to the xanthine-xanthine oxidase ROS generating system significantly reduced both SL kinase activities. Superoxide dismutase did not prevent this inhibition; however, catalase significantly prevented the xanthine-xanthine oxidase induced inhibition. Treatment of SL with hydrogen peroxide (H2O2) resulted in inhibition of both the kinases, which was prevented by catalase and dithiothreitol (DTT). Hypochlorous acid also inhibited both the kinases, which was prevented by DTT. Deferoxamine (an iron chelator) and mannitol (an *OH scavenger) did not modify the H2O2-induced depression of the kinases, eliminating any role of *OH. Furthermore, the IC50 of H2O2 on PtdIns 4 kinase and PtdIns 4-P 5-kinase was 27 and 81 microM, respectively. In addition, inclusion of reduced glutathione in the assay of the kinases in the absence of H2O2 did not affect the activities of the kinases; however, oxidized glutathione induced a significant depression. Also, a significant decline of the PtdIns 4 kinase and PtdIns 4-P 5-kinase activities due to changing of the redox ratio was observed. Thiol modifiers (N-ethylmaleimide, methyl methanethiosulfonate, or p chloromercuriphenylsulfonic acid) were detected to depress the kinases' activities, which were substantially prevented by DTT. The results suggest that functionally critical thiol groups may be associated with PtdIns 4-kinase and PtdIns 4-P 5-kinase and that changes of their redox state by ROS can impair their activities, which may be an important factor in the oxidant-induced cardiac dysfunction. PMID- 11051097 TI - Thermodynamics of unfolding of beta-trypsin at pH 2.8. AB - The unfolding equilibrium of beta-trypsin induced by thermal and chemical denaturation was thermodynamically characterized. Thermal unfolding equilibria were monitored using UV absorption and both far- and near-UV CD spectroscopy, while fluorescence was used to monitor urea-induced transitions. Thermal and urea transition curves are reversible and cooperative and both sets of data can be reasonably fitted using a two-state model for the unfolding of this protein. Plots of the fraction denatured, calculated from thermal denaturation curves at different wavelengths, versus temperature are coincident. In addition, the ratio of the enthalpy of denaturation obtained by scanning calorimetry to the van't Hoff enthalpy is close to unity, which supports the two-state model. Considering the differences in experimental approaches, the value for the stability of beta trypsin estimated from spectroscopic data (deltaGu = 6.0 +/- 0.2 kcal/mol) is in reasonable agreement with the value calculated from urea titration curves (deltaGUH2O = 5.5 +/- 0.3 kcal/mol) at pH 2.8 and 300 degrees K. PMID- 11051098 TI - Function of glutathione peroxidase in endothelial cell vitality. AB - The two human umbilical vein endothelial cell-derived lines, ECRF24 and ECV304, differ in responsiveness to oxidative stress. In confluent monolayers of ECRF24, but not in ECV304, peroxides induce stress responses such as plasma membrane blebbing and nuclear condensation. The peroxide effect on ECRF24 was preceded by oxidation of reduced glutathione (GSH) and of NAD(P)H, and by oxidation of the redox-sensitive probe, chloromethyl 2',7'-dichlorofluorescin (DCFH). In monolayers of ECV304, peroxides induced only minimal oxidation of GSH, NAD(P)H and DCFH, which was associated with a greatly reduced GSH peroxidase activity in these cells. However, in spite of the absence of a blebbing response, ECV304 were more susceptible than ECRF24 to membrane lipid peroxidation and peroxide-induced necrosis. Only for ECV304, the culturing with high levels of polyunsaturated fatty acids increased lipid peroxidation and cellular death. Treatment of these cells with the GSH peroxidase mimic ebselen effectively reversed their decreased vitality. We conclude that, in peroxide-treated endothelial cells, cell death (necrosis) can result from lipid peroxidation by peroxide that has not been removed by GSH peroxidases, whereas extensive peroxidase activity may cause a stress response (blebbing). The data further identify ECV304 as a stress sensitive cell line, where peroxides exert their effects independently of GSH oxidation. PMID- 11051099 TI - Effects of metal ions on the activity of protein tyrosine phosphatase VHR: highly potent and reversible oxidative inactivation by Cu2+ ion. AB - The posttranslational regulation of protein tyrosine phosphatases (PTPs) has been suggested to have a crucial role in maintaining the phosphotyrosine level in cells. Here we examined the regulatory effects of metal ions on human dual specificity vaccinia H1-related protein tyrosine phosphatase (VHR) in vitro. Among various metal ions examined, Fe3+, Cu2+, Zn2+, and Cd2+ exerted their inactivational effects on VHR, and Cu2+ is the most potent inactivator. The VHR activity inactivated by the metal ions except Cu2+ was significantly restored by EDTA. The efficacy of Cu2+ for the VHR inactivation was about 200-fold more potent than that of H2O2. Cu2+ also inactivated other PTPs including PTP1B and SHP-1. The Cu2+-mediated inactivation at the submicromolar range was eradicated by dithiothreitol treatment. The loss of VHR activity correlated with the decreased [14C]iodoacetate labeling of active-site cysteine, suggesting that Cu2+ brought about the oxidation of the active-site cysteine. On the contrary, Zn2+ that exerted an inactivational effect at millimolar concentrations appeared not directly linked to the active-site cysteine, as indicated by the fact that [14C]iodoacetate labeling was unaffected and that the effect of Zn2+ on the Y78F mutant was increased. The reduction potential of VHR was estimated to be -331 mV by utilizing the reversibility of the redox state of VHR. Thus, we conclude that the highly potent Cu2+ inactivation of VHR is a consequence of the oxidation of the active-site cysteine and the mode of Zn2+ inactivation is distinct from that of Cu2+. PMID- 11051100 TI - Cadmium-induced mRNA expression of Hsp32 is augmented in metallothionein-I and II knock-out mice. AB - Cadmium is toxic and carcinogenic to humans and animals. The testis and lung are the target organs for cadmium carcinogenesis. Heat shock proteins (HSPs) as well as metallothionein (MT) and glutathione (GSH) play an important role in protection against its toxicity. HSP32, also known as heme oxygenase-1, is a 32 kDa protein induced by heme, heavy metals, oxidative stresses, and heat. We investigated expression of the Hsp32 gene of various organs (the liver, lung, heart, stomach, kidney, and testis) in transgenic mice deficient in the MT-I and II genes (MT-KO) and in control mice (MT-W) after an injection of cadmium chloride (CdCl2). Survival of MT-W mice after a subcutaneously injection of CdCl2 was higher than that of MT-KO mice, while no significant difference was observed in the level of GSH in each organ between MT-W and MT-KO mice. Northern blot analysis showed that the MT-I mRNA was more extensively induced in the liver, kidney, and heart than other organs 6 h after an injection of CdCl2 (30 micromol/kg body wt, sc). There was little increase of the MT-I mRNA in the testis when induced by CdCl2. Expression of the Hsp32 gene in the liver and kidney in response to CdCl2 was more extensively augmented in MT-KO mice than in MT-W mice. In the lung and testis, there was little induction and no augmentation in expression of the Hsp32 gene induced by CdCl2 in both MT-W and MT-KO mice. In the stomach, there was little induction of the Hsp32 mRNA in MT-W mice, but was increased in MT-KO mice. Immunohistochemical staining revealed that the HSP32 protein was strongly expressed in the kidney and liver of MT-W mice 24 h after an injection of CdCl2 (20 micromol/kg body wt, sc), while the expression of HSP32 protein was not increased in the testis. In metabolically active organs such as the liver and kidney, expression of the Hsp32 gene as well as the MT-I gene was extensively induced by cadmium in MT-W mice, and more eminently induced in MT-KO mice. We suggest that organs of low stress response to cadmium such as the testis and lung may be vulnerable target sites for cadmium toxicity and carcinogenesis. PMID- 11051101 TI - Reactivity of manganese peroxidase: site-directed mutagenesis of residues in proximity to the porphyrin ring. AB - The purpose of this study was to determine the effect of heme pocket hydrophobicity on the reactivity of manganese peroxidase. Residues within 5 A of the heme active site were identified. From this group, Leu169 and Ser172 were selected and mutated to Phe and Ala, respectively. The mutant proteins were then characterized by steady-state kinetics. Whereas the Leu169Phe mutation had little, if any, effect on activity, the Ser172Ala mutation decreased kcat and also the specificity constant (kcat/Km) for Mn2+, but not H2O2. Transient-state studies indicated that the mutation affected only the reactions of compound II. These results indicate that compound II is the most sensitive to changes in the heme environment. PMID- 11051102 TI - Redox control of aryl sulfotransferase specificity. AB - Aryl sulfotransferase IV from rat liver has the very broad substrate range that is characteristic of the enzymes of detoxication. With the conventional assay substrates, 4-nitrophenol and PAPS, sulfation was considered optimal at pH 5.5 whereas the enzyme in the physiological pH range was curiously ineffective. These properties would seem to preclude a physiological function for this cytosolic enzyme. Partial oxidation of the enzyme, however, results not only in a substantial increase in the rate of sulfation of 4-nitrophenol at physiological pH but also in a shift of the pH optimum to this range and radically altered overall substrate specificity. The mechanism for this dependence on redox environment involves oxidation at Cys66, a process previously shown to occur by formation of a mixed disulfide with glutathione or by the formation of an internal disulfide with Cys232. Oxidation at Cys66 acts only as a molecular redox switch and is not directly part of the catalytic mechanism. Underlying the activation process is a change in the nature of the ternary complex formed between enzyme, phenol, and the reaction product, adenosine 3',5'-bisphosphate. The reduced enzyme gives rise to an inhibitory, dead-end ternary complex, the stability of which is dictated by the ionization of the specific phenol substrate. Ternary complex formation impedes the binding of PAPS that is necessary to initiate a further round of the reaction and is manifest as profound, substrate-dependent inhibition. In contrast, the ternary complex formed when the enzyme is in the partially oxidized state allows binding of PAPS and the unhindered completion of the reaction cycle. PMID- 11051103 TI - Site-directed mutagenesis improves catalytic efficiency and thermostability of Escherichia coli pH 2.5 acid phosphatase/phytase expressed in Pichia pastoris. AB - Escherichia coli pH 2.5 acid phosphatase gene (appA) and three mutants were expressed in Pichia pastoris to assess the effect of strategic mutations or deletion on the enzyme (EcAP) biochemical properties. Mutants A131N/ V134N/D207N/S211N, C200N/D207N/S211N, and A131N/ V134N/C200N/D207N/S211N had four, two, and four additional potential N-glycosylation sites, respectively. Extracellular phytase and acid phosphatase activities were produced by these mutants and the intact enzyme r-AppA. The N-glycosylation level was higher in mutants A131N/V134N/D207N/S211N (48%) and A131N/V134N/ C200N/D207N/S211N (89%) than that in r-AppA (14%). Despite no enhancement of glycosylation, mutant C200N/ D207N/S211N was different from r-AppA in the following properties. First, it was more active at pH 3.5-5.5. Second, it retained more (P < 0.01) phytase activity than that of r-AppA. Third, its specific activity of phytase was 54% higher. Lastly, its apparent catalytic efficiency kcat/Km for either p-nitrophenyl phosphate (5.8 x 10(5) vs 2.0 x 10(5) min(-1) M(-1)) or sodium phytate (6.9 x 10(6) vs 1.1 x 10(6) min(-1) M(-1)) was improved by factors of 1.9- and 5.3-fold, respectively. Based on the recently published E. coli phytase crystal structure, substitution of C200N in mutant C200N/D207N/S211N seems to eliminate the disulfide bond between the G helix and the GH loop in the alpha-domain of the protein. This change may modulate the domain flexibility and thereby the catalytic efficiency and thermostability of the enzyme. PMID- 11051104 TI - The folding of nascent mitochondrial aspartate aminotransferase synthesized in a cell-free extract can be assisted by GroEL and GroES. AB - At 30 degrees C, the precursor to mitochondrial aspartate aminotransferase (pmAspAT) cannot fold after synthesis in rabbit reticulocyte lysate (RRL), a model for studying intracellular protein folding. However, it folds rapidly once imported into mitochondria. Guanidinium chloride denatured pmAspAT likewise cannot refold at 30 degrees C in a defined in vitro system. However, it refolds rapidly and in good yield in the presence of the intramitochondrial chaperone homologues GroEL and GroES. In this report, we demonstrate that GroEL and GroES can also facilitate the folding of nascent pmAspAT in reticulocyte lysate under conditions where it otherwise would not. When added alone, GroEL arrests the slow folding of nascent pmAspAT and inhibits import into mitochondria. These effects are significantly reversed by adding GroES. These observations suggest that added GroEL participates in an equilibrium with endogenous chaperones in the cytosol which inhibit folding and promote import competence. Native gel electrophoresis suggests that nascent pmAspAT exists in RRL as a heterogeneous population of partially folded species, some of which bind to added GroEL more readily than others. The GroEL-trapped species appear to be among the productive pmAspAT folding intermediates formed in RRL or they at least appear to equilibrate with these intermediates, since they become import competent after GroES-stimulated release from GroEL. PMID- 11051105 TI - Inhibition of prothrombin activation by bothrojaracin, a C-type lectin from Bothrops jararaca venom. AB - Bothrojaracin is a potent and specific alpha-thrombin inhibitor (Kd approximately 0.6 nM) isolated from Bothrops jararaca venom. It binds to both of thrombin's anion-binding exosites (1 and 2), thus inhibiting the ability of the enzyme to act upon several natural macromolecular substrates, such as fibrinogen, platelet receptor, protein C, and factor V. Additionally, bothrojaracin interacts with prothrombin (Kd approximately 30 nM), as previously determined by a solid-phase assay. However, there is no information concerning the effect of this interaction on prothrombin activation and whether the binding of bothrojaracin can occur in plasma. Here, we show that bothrojaracin specifically interacts with prothrombin in human plasma. It is an effective anticoagulant after activation of the intrinsic pathway of blood coagulation, and analysis of prothrombin conversion in plasma shows that bothrojaracin strongly reduces alpha-thrombin formation. To determine whether this effect is due exclusively to inhibition of feedback reactions involving the thrombin-induced activation of factors V and VIII, we analyzed the effect of bothrojaracin on the activation of purified prothrombin by Oxyuranus scutellatus venom. As with plasma, bothrojaracin greatly inhibited thrombin formation, suggesting a direct interference in the prothrombin activation by the enzyme found in this venom (scuterin, a prothrombin activator described as a factor Xa/factor Va-like complex). Altogether, we suggest that bothrojaracin exerts its anticoagulant effect in plasma by two distinct mechanisms: (1) it binds generated thrombin and inhibits exosite 1 dependent activities such as fibrinogen clotting and factor V activation, and (2) it interacts with prothrombin and decreases its proteolytic activation. Thus, bothrojaracin may be useful in the search for thrombin inhibitors that bind both the zymogen and the active enzyme. PMID- 11051106 TI - Promoter function and the role of cytokines in the transcriptional regulation of rabbit CYP2E1 and CYP2E2. AB - Rabbit CYP2E1 and CYP2E2 show considerable similarity in the 5' flanking region, but a 32-base-pair element (32-BPE) that is repeated in 2E1 is present only as a single inexact copy in 2E2. In the present investigation, footprinting disclosed two specific binding sites for liver nuclear proteins, and the DNase I sensitivity profiles of the two genes were found to be different. Several positive and negative regulatory elements were identified by transfection with a series of constructs of upstream CYP2E sequences fused to the luciferase gene. Both genes have an HNF-1 consensus motif with one nucleotide mismatch, which affects binding affinity and promoter activity. Investigation of DNA-protein interactions revealed that Sp1 and NFkappaB bind exclusively to the 32-BPE of 2E1 and 2E2, respectively, suggesting a possible regulatory role for the 32-BPE. Interleukin-1alpha (IL-1alpha) gave rise to a 2.5-fold increase in the promoter activity of 2E1 in HepG2 cells, and the IL-1alpha-mediated induction of reporter gene expression was almost completely prevented when the 32-BPE was deleted. Increased DNA binding and Sp1 protein content as a result of IL-1alpha treatment, as well as cotransfection experiments with pPacSp1, suggest that Sp1 is a transcription activator for the induction of 2E1 by IL-1alpha in HepG2 cells. PMID- 11051107 TI - Use of pH and kinetic isotope effects to dissect the effects of substrate size on binding and catalysis by nitroalkane oxidase. AB - The flavoprotein nitroalkane oxidase catalyzes the oxidation of a broad range of primary and secondary nitroalkanes to the respective aldehydes or ketones, with production of hydrogen peroxide and nitrite. The V/K values for primary nitroalkanes increase with increasing chain length, reaching a maximum with 1 nitrobutane [Gadda, G., and Fitzpatrick, P. F. (1999) Arch. Biochem. Biophys. 363, 309-313]. In the present report, pH and deuterium kinetic isotope effects with a series of primary nitroalkanes and phenylnitromethane as substrates have been used to dissect the effects of chain length on binding and catalysis. The apparent pKa value for a group that must be unprotonated for catalysis decreases from about 7 to 5.3 with increasing size of the substrate. The D(V/K) values for these substrates decrease from 7.5 with nitroethane to 1 with phenylnitromethane. These results show that increasing the size of the substrate results in an increased partitioning forward to catalysis. The D(V/K) and DVmax values at pH 5.5 have been used to calculate the effect of substrate size on the Kd values for primary nitroalkanes. The Kd values decrease with increasing length of the substrate, with a deltadeltaG(binding) of 1.7 kcal mol(-1) for each additional methylene group. Such a value is less than the value of 2.6 kcal mol(-1) previously determined for the effect of a methylene group on the V/K value [Gadda, G., and Fitzpatrick, P. F. (1999) Arch. Biochem. Biophys. 363, 309-313], suggesting that the total energy available per methylene group is used not only to enhance binding but also to increase the rate of catalysis. PMID- 11051108 TI - Purification and characterization of S-adenosyl-L-methionine:benzoic acid carboxyl methyltransferase, the enzyme responsible for biosynthesis of the volatile ester methyl benzoate in flowers of Antirrhinum majus. AB - S-Adenosyl-L-methionine:benzoic acid carboxyl methyltransferase (BAMT) catalyzes the transfer of the methyl group of S-adenosyl-L-methionine (SAM) to the carboxyl group of benzoic acid to make the volatile ester methyl benzoate, one of the most abundant scent compounds of snapdragon, Antirrhinum majus. The enzyme was purified from upper and lower petal lobes of 5- to 10-day-old snapdragon flowers using DE53 anion exchange, Phenyl-Sepharose 6FF, and Mono-Q chromatography. The purified protein has a pH optimum of 7.5 and is highly specific for benzoic acid, with no activity toward several other naturally occurring substrates such as salicylic acid, cinnamic acid, and their derivatives. The molecular mass values for native and denatured protein were 100 and 49 kDa, respectively, suggesting that the active enzyme is a homodimer. The addition of monovalent cations K+ and NH4+ stimulates BAMT activity by a factor of 2, whereas the addition of Fe2+ and Cu2+ has a strong inhibitory effect. Plant-purified BAMT has Km values of 28 microM and 1.1 mM for SAM and benzoic acid, respectively (87 microM and 1.6 mM, respectively, for plant BAMT expressed in Escherichia coli). Product inhibition studies showed competitive inhibition between SAM and S-adenosyl-L-homocysteine (SAH), with a Ki of 7 microM, and noncompetitive inhibition between benzoic acid and SAH, with a Ki of 14 microM. PMID- 11051109 TI - Ectonucleotide diphosphohydrolase activities in hemocytes of larval Manduca sexta. AB - In this work, we describe the ability of living hemocytes from an insect (Manduca sexta, Lepidoptera) to hydrolyze extracellular ATP. In these intact cells, there was a low level of ATP hydrolysis in the absence of any divalent metal (8.24 +/- 0.94 nmol of Pi/h x 10(6) cells). The ATP hydrolysis was stimulated by MgCl2 and the Mg2+-dependent ecto-ATPase activity was 15.93 +/- 1.74 nmol of Pi/h x 10(6) cells. Both activities were linear with cell density and with time for at least 90 min. The addition of MgCl2 to extracellular medium increased the ecto-ATPase activity in a dose-dependent manner. At 5 mM ATP, half-maximal stimulation of ATP hydrolysis was obtained with 0.33 mM MgCl2. This stimulatory activity was not observed when Ca2+ replaced Mg2+. The apparent Km values for ATP-4 and Mg-ATP2- were 0.059 and 0.097 mM, respectively. The Mg2+-independent ATPase activity was unaffected by pH in the range between 6.6 and 7.4, in which the cells were viable. However, the Mg2+-dependent ATPase activity was enhanced by an increase of pH. These ecto-ATPase activities were insensitive to inhibitors of other ATPase and phosphatase activities, such as oligomycin, sodium azide, bafilomycin A1, ouabain, furosemide, vanadate, sodium fluoride, tartrate, and levamizole. To confirm the observed hydrolytic activities as those of an ecto-ATPase, we used an impermeant inhibitor, DIDS (4,4'-diisothiocyanostilbene-2,2'-disulfonic acid), as well as suramin, an antagonist of P2-purinoreceptors and inhibitor of some ecto ATPases. These two reagents inhibited the Mg2+-independent and the Mg2+-dependent ATPase activities to different extents. Interestingly, lipopolysaccharide, a component of cell walls of gram-negative bacteria that increase hemocyte aggregation and phagocytosis, increased the Mg2+-dependent ecto-ATPase activity in a dose-dependent manner but did not modify the Mg2+-independent ecto-ATPase activity. PMID- 11051110 TI - Viruses and cancer. PMID- 11051111 TI - Oxychlordane and trans-nonachlor in breast adipose tissue and risk of female breast cancer. AB - BACKGROUND: Organochlorine compounds, including organochlorine pesticides, have been suggested by some, but not all, studies to be associated with female breast cancer risk. So far, studies relating organochlorine compounds and breast-cancer risk have mainly focused on polychlorinated biphenyls (PCBs) and dichlorodiphenyltrichloroethane (DDT) as risk factors for female breast cancer. This paper examines the hypothesis that environmental exposure to trans-nonachlor (TNC) and oxychlordane (OCD), a major metabolite of the insecticide chlordane, increases the METHODS: A total of 304 histologically confirmed, incident primary breast-cancer patients and 186 histologically confirmed incident benign breast disease controls were included in the study between 1994 and 1997. Breast adipose tissue not needed for diagnostic purposes was collected and analysed for TNC, OCD and other organochlorine compounds. A standardised, structured questionnaire was used to obtain information on major known, or suspected, risk factors for breast cancer. RESULTS: The age and lipid-adjusted geometric mean adipose-tissue levels of OCD were similar between the cases [36.4 p.p.b., 95% confidence interval (CI) 34.7-38.2 p.p.b.] and controls (38.0 p.p.b., 95% Cl 35.7-40.6 p.p.b.). The age and lipid-adjusted geometric mean adipose-tissue levels of TNC between the cases (55.5 p.p.b., 95% CI 52.6-58.5 p.p.b.) and controls (58.1 p.p.b., 95% CI 54.2 62.3 p.p.b.) were also similar. There was no association between breast-cancer risk and mean adipose-tissue levels of OCD and TNC. The covariate-adjusted odds ratio (OR) was 0.7 (95% CI 0.4-1.3) for OCD and 1.1 (95% CI 0.6-1.9) for TNC, when the highest quartile was compared with the lowest. The risk also did not vary based on oestrogen or progesterone receptor status or menopausal status. DISCUSSION: We found no significantly increased risk of breast cancer associated with breast adipose-tissue levels of OCD or TNC; this is consistent with recent epidemiological studies, indicating that environmental exposure to organochlorine compounds does not have an overall significant impact on breast-cancer risk. PMID- 11051112 TI - Estimation of a time-varying force of infection and basic reproduction number with application to an outbreak of classical swine fever. AB - BACKGROUND: A method was developed for stochastically reconstructing the pattern of infection from observed epidemic data. This allowed for estimation of a time dependent force of infection, or transmission rate, during an epidemic. METHODS: A discrete-time mechanistic model was used to describe the spread of infection and a stochastic procedure, which utilised the latent and infectious period distributions, was used to reconstruct the dates of infection, becoming infectious and removal from the given data. The four equations describing the model were then solved to obtain least squares estimates of the transmission rate and the basic reproduction number (R0) throughout the epidemic. This process was repeated in order to assess the variability in these key epidemiological parameters. The stochastic epidemic reconstruction procedure was developed to account for changes in the distribution of the survival period over the course of the epidemic and adapted for application to epidemic data where not all infected individuals have yet been observed as cases. RESULTS: The method was applied to a set of epidemic data from an outbreak of classical swine fever in Pakistan. Constant and time-varying estimates of the transmission rate were derived and compared. There was some evidence to suggest that the force of infection varied over time. DISCUSSION: The method described can be applied to data from epidemics where observations are incomplete. The confidence limits obtained for the estimated force of infection provide a means of assessing the evidence for time variation in this parameter. PMID- 11051113 TI - The use of frailty models in genetic studies: application to the relationship between end-stage renal failure and mutation type in Alport syndrome. European Community Alport Syndrome Concerted Action Group (ECASCA). AB - BACKGROUND: Alport syndrome (AS) is a severe hereditary disease usually transmitted as an X dominant trait and involving a mutation of the COL4A5 gene. It leads to end-stage renal failure (ESRF), but this progression is heterogeneous. Mutations of the COL4A5 gene have been characterised in numerous families using molecular biology. Our objective was to evaluate the interfamilial heterogeneity of the disease and to study relationships between mutation types and progression to ESRF in the European Community Alport Syndrome Concerted Action group (ECASCA) registry database. METHODS: We used the frailty model framework. Frailty models have been developed to analyse censored data with non independent observations. Random effects are introduced in a Cox proportional regression model to take into account the intracluster correlations. In this study, ESRF is considered a censored event and the intrafamilial correlations are taken into account in the frailty models. RESULTS: These approaches allow us to demonstrate the existence of an interfamilial heterogeneity; the role of the mutation type explains the interfamilial variability. In particular, the results suggest that some mutation types are associated with a higher risk of ESRF for males. CONCLUSIONS: This study shows the importance of characterising the mutation at the molecular level in genetic studies, to understand the relationship between genotype and phenotype. The frailty models constitute an attractive approach in this context, when the phenotype is characterised by a censored end-point. PMID- 11051114 TI - Nonparametric evaluation of birth cohort trends in disease rates. AB - BACKGROUND: Although interpretation of age-period-cohort analyses is complicated by the non-identifiability of maximum likelihood estimates, changes in the slope of the birth-cohort effect curve are identifiable and have potential aetiologic significance. METHODS: A nonparametric test for a change in the slope of the birth-cohort trend has been developed. The test is a generalisation of the sign test and is based on permutational distributions. A method for identifying interactions between age and calendar-period effects is also presented. RESULTS: The nonparametric method is shown to be powerful in detecting changes in the slope of the birth-cohort trend, although its power can be reduced considerably by calendar-period patterns of risk. The method identifies a previously unidentified decrease in the birth-cohort risk of lung-cancer mortality from 1912 to 1919, which appears to reflect a reduction in the initiation of smoking by young men at the beginning of the Great Depression (1930s). The method also detects an interaction between age and calendar period in leukemia mortality rates, reflecting the better response of children to chemotherapy. CONCLUSION: The proposed nonparametric method provides a data analytic approach, which is a useful adjunct to log-linear Poisson analysis of age-period-cohort models, either in the initial model building stage, or in the final interpretation stage. PMID- 11051115 TI - Comparing methods for estimating the variation of risks of cancer between small areas. AB - BACKGROUND: Analysing the geographical variation of cancer incidence is an important issue in epidemiological research. It might suggest new aetiologic hypotheses, provide guidelines for the design of new surveys and give ideas for preventive campaigns. METHODS: Four different methods for estimating the variation of cancer risks between small areas and three homogeneity tests were evaluated by simulation. In three of the methods the systematic variation of the relative risks (RR) was estimated by subtracting the expected Poisson variation from the total variation. The last method assumes that RR are gamma distributed and the maximum likelihood estimate (MLH) of the systematic variation parameter is calculated. A likelihood ratio test (LRT) of heterogeneity of RR based on this method is also evaluated, and compared with an ordinary chi2 test and the Potthoff and Whittinghill test (P&W). RESULTS: For most of the simulated data sets, the estimates obtained by MLH are most precise, even if the assumption of gamma distribution of RR is violated. The LRT and P&W tests of homogeneity are also shown to perform better than the chi2 test. Most of the real cancer data sets exhibited at least some geographical variation. Cancer of the lung, melanoma and other skin cancers, and cancers of the urinary bladder, pancreas and stomach, have the highest systematic variation. DISCUSSION: The study suggests that likelihood-based approaches are suitable, both for estimating the variation between small areas and for testing the null hypothesis of equal RR. Such geographical analyses might suggest new aetiological hypothesis. PMID- 11051116 TI - A 'sufficient cause' model for dental caries. AB - BACKGROUND: It is generally believed that dental caries is an infectious disease. The occurrence of dental caries is affected by a variety of determinants. In order to estimate the precise extent of the relation between specific determinants and the outcome phenomenon (i.e. the occurrence of dental caries), a coherent disease model is required. This model should also permit multivariate analysis to control for confounders and interactions. Only with such a disease model will it be possible to investigate the relation between the occurrence of a determinant and dental caries, and to estimate the extent of this relation. The known causal models for the explanation of dental caries do not fully meet these requirements. METHOD: Rothman's 'sufficient cause' model has been used as a starting point for the development of a new coherent disease model, to explain the occurrence of dental caries and allow multivariate analysis. RESULTS: The sufficient cause for dental caries comprises three component causes: sufficient microorganisms with cariogenic potential, easily fermentable carbohydrates and teeth. Whether dental caries actually occurs also depends on the influence of independent risk factors that interact with the component causes in a protective, as well as in a risk-increasing manner. These independent risk factors are saliva, fluoride, oral hygiene and diet. CONCLUSIONS: The 'sufficient cause' model for dental caries is a biological model in which distinction between protective and risk-increasing factors has been made, and interaction between factors has been described. With this model, it will now be possible to assess the extent of the relationship between a determinant and dental caries (the outcome phenomenon) using multivariate techniques. PMID- 11051117 TI - Anticonvulsant action of topiramate against motor seizures in developing rats. AB - PURPOSE: To study the anticonvulsant action of topiramate (TPM) in developing rats. METHODS: Motor seizures were elicited by administering pentylenetetrazol (100 mg/kg subcutaneously) in five age groups of Wistar rats (7, 12, 18, 25, and 90 days old). TPM was administered intraperitoneally in doses from 10 to 640 mg/kg 2 hours before pentylenetetrazol. The time course of TPM action was studied in 12- and 25-day-old rats up to 24 hours after the 160-mg/kg dose, and the incidence and pattern of seizures were evaluated. RESULTS: TPM did not influence minimal seizures (clonus of forelimb and head muscles with preserved righting ability). Generalized tonic-clonic seizures, however, were reliably changed at all developmental stages studied. The tonic phase was suppressed so that the majority of animals exhibited generalized clonic seizures (with a loss of righting reflexes). In addition, the incidence of generalized seizures was decreased after the 20-, 40-, and 80-mg/kg doses in the 7-day-old rat pups. The specific suppression of the tonic phase of generalized seizures was observed up to 12 hours in the 12-day-old rat pups. The same result was obtained over 6 hours after TPM administration in the 25-day-old animals, and with longer intervals the incidence of generalized seizures decreased in this age group. CONCLUSIONS: TPM exhibits stable anticonvulsant action against the tonic phase of generalized tonic-clonic seizures throughout development. In addition, it suppresses all phases of generalized seizures in 7-day-old rats. The anticonvulsant action of TPM lasted longer in 25-day-old than in 12-day-old rats. The two actions of TPM might be ascribed to two different mechanisms of action. PMID- 11051118 TI - Rapid cooling aborts seizure-like activity in rodent hippocampal-entorhinal slices. AB - PURPOSE: As a preliminary step in the development of an implantable Peltier device to abort focal neocortical seizures in vivo, we have examined the effect of rapid cooling on seizures in rodent hippocampal-entorhinal slices. METHODS: Seizure-like discharges were induced by exposing the slices to extracellular saline containing 4-aminopyridine (50 micromol/L). RESULTS: When we manually activated a Peltier device that was in direct contact with the slice, seizures terminated within seconds of the onset of cooling, sometimes preceding a detectable decrease in temperature measured near the top of the slice. However, activation of the Peltier device did not stop seizures when slices were no longer in direct physical contact with the device, indicating that this was not a field effect. When cooling was shut off and temperature returned to 33 degrees C, bursting sometimes returned, but a longer-term suppressive effect on seizure activity could be observed. In two of our experiments, a custom computer program automatically detected seizure discharges and triggered a transistor-transistor logic pulse to activate the Peltier device. In these experiments, the Peltier device automatically terminated the slice bursting in less than 4 seconds. When the Peltier device was placed in contact with the normal, exposed cortex of a newborn pig, we found that the cortical temperature decreased rapidly from 36 degrees C to as low as 26 degrees C at a depth of 1.7 mm below the cooling unit. CONCLUSIONS: These experiments show that local cooling may rapidly terminate focal paroxysmal discharges and might be adapted for clinical practice. PMID- 11051119 TI - Modulation of calcium channels by group I and group II metabotropic glutamate receptors in dentate gyrus neurons from patients with temporal lobe epilepsy. AB - PURPOSE: Metabotropic glutamate receptors (mGluRs) might be promising new drug targets for the treatment of epilepsy because the expression of certain mGluRs is regulated in epilepsy and because activation of mGluRs results in distinctive anti- and proconvulsant effects. Therefore, we examined how mGluR activation modulates high-voltage-activated (HVA) Ca2+ channels. METHODS: Whole-cell patch clamp recordings were obtained from granule cells and interneuron-like cells acutely isolated from the dentate gyrus of patients with pharmacoresistent temporal lobe epilepsy. RESULTS: Agonists selective for either group I or group II mGluRs rapidly and reversibly reduced HVA currents in most dentate gyrus cells. These modulatory effects were inhibited by the respective group I and group II mGluR antagonists. The specific Ca2+ channel antagonists nifedipine and omega-conotoxin GVIA potently occluded the effects of group I and II mGluR agonists, respectively, indicating that group I mGluRs acted on L-type channels and group II mGluRs affected N-type channels. About two thirds of the responsive neurons were sensitive either to group I or group II mGluRs, whereas a minority of cells showed effects to agonists of both groups, indicating a variable mGluR expression pattern. CONCLUSIONS: Group I and group II mGluRs are expressed in human dentate gyrus neurons and modulate L- and N-type HVA channels, respectively. The data shed light on the possible cellular sequelae of the mGluR1 upregulation observed in human epileptic dentate gyrus as well as on possible mGluR-mediated anticonvulsant mechanisms. PMID- 11051120 TI - Nitric oxide synthase expression in the cerebral cortex of patients with epilepsy. AB - PURPOSE: Nitric oxide (NO), a short-lived radical synthesized from L-arginine by activation of the enzyme nitric oxide synthase (NOS), has been implicated in the pathophysiology of epilepsy by some investigators. However, the current data about NO and NOS in epilepsy are controversial and are derived only from animal models of epilepsy. In this study we investigated possible changes in NOS expression in the cerebral cortex of patients with epilepsy. METHODS: Qualitative and quantitative parameters of the immunolabeling pattern of the neuronal, endothelial, and inducible isoforms of NOS were analyzed in biopsy material obtained from patients with short and long seizure history and from patients without epilepsy. RESULTS: The comparative study showed that in the cerebral cortex of patients with epilepsy, particularly in those with a long seizure history, the number and labeling intensity of NOS-positive neurons increased, and that a subpopulation of nonpyramidal GABAergic neurons (type II NOS neurons) was responsible for this phenomenon. CONCLUSIONS: The fact that NOS upregulation is more evident in patients with a long seizure history suggests that this is a consequence of seizures, acting probably as an adaptative response to the sustained release of excitatory amino acids. PMID- 11051121 TI - How well can epilepsy syndromes be identified at diagnosis? A reassessment 2 years after initial diagnosis. AB - PURPOSE: Epilepsy syndromes can be identified very early in the course of a seizure disorder. It is unclear how accurate and resilient such early classifications are. We compared the classification of epilepsy syndromes made previously on the basis of information available at diagnosis with those made 2 years later in a cohort of children with newly diagnosed epilepsy. METHODS: Children (n = 613) were prospectively identified at the time of initial diagnosis by participating physicians in Connecticut between 1993 and 1997. Classification of epilepsy syndrome according to International League Against Epilepsy guidelines was made previously based on all relevant information available at diagnosis. All cases were reclassified again after 2 years of additional evidence had accumulated. The distributions of syndromes at diagnosis and at 2 years are compared and reasons for changes examined. RESULTS: After 2 years, syndromes remained the same in 86.3% of the cohort and changes occurred in 13.7% (n = 84). Evolution of the syndrome occurred in 24 children (3.9%), and rectification to the initial diagnosis occurred in 60 children (9.8%). The most common scenario for evolution of a syndrome was from West syndrome (n = 5), undetermined (n = 4), or symptomatic localization-related epilepsy (n = 3) to the Lennox-Gastaut syndrome. The most common rectification of initial classifications involved incompletely classified syndromes (cryptogenic localization-related and undetermined syndromes; n = 36). In a few instances, a fully specified syndrome was reclassified to another apparently unrelated syndrome. In these cases, initial information at diagnosis had been difficult to interpret. CONCLUSIONS: Epilepsy syndromes can, for the most part, be identified at the time of initial diagnosis. Two years later, rectifications were made in only 9.8% of cases, and most of these involved syndromes that represented incomplete classifications in the first place. Significant changes were rare. The International League Against Epilepsy classification of the epilepsies can be meaningfully applied in epidemiological studies of newly diagnosed pediatric epilepsy. PMID- 11051122 TI - Efficacy and tolerability of levetiracetam 3000 mg/d in patients with refractory partial seizures: a multicenter, double-blind, responder-selected study evaluating monotherapy. European Levetiracetam Study Group. AB - PURPOSE: To evaluate the efficacy and tolerability of levetiracetam (LEV) monotherapy in selected patients with refractory partial seizures. METHODS: In this multicenter, double-blind, placebo-controlled, parallel-group, responder selected study, patients were randomized (2:1 ratio) to receive oral LEV 1500 mg twice daily or placebo during a 12-week add-on phase. Treatment responders (patients with a reduction in partial seizure frequency of 50% or more compared with baseline) entered a monotherapy phase that included a maximum 12-week down titration period and 12 weeks of monotherapy at 1500 mg twice daily. In both phases, responder rate, seizure frequency, and adverse events were analyzed. RESULTS: A total of 286 patients (placebo, n = 105; LEV, n = 181) entered the add on phase, and 86 patients (placebo, n = 17; LEV, n = 69) were eligible for the monotherapy phase. Thirty-six of 181 patients (19.9%) who received LEV completed the entire study compared with only 10 of 105 patients (9.5%) in the placebo group (p = 0.029). The odds of completing the study on LEV were 2.36 times (95% confidence interval, 1.08, 5.57) higher than on placebo. The responder rate during the add-on phase was significantly higher in the LEV group compared with the placebo group (42.1% vs. 16.7%, respectively; p < 0.001). In the LEV monotherapy group, the median percent reduction in partial seizure frequency compared with baseline was 73.8% (p = 0.037), with a responder rate of 59.2%. Nine patients (18.4%) remained seizure-free on LEV monotherapy. CONCLUSIONS: Conversion to LEV monotherapy (1500 mg twice daily) is effective and well tolerated in patients with refractory partial seizures who responded to 3000 mg/d LEV as add-on therapy. PMID- 11051123 TI - Sulthiame as monotherapy in children with benign childhood epilepsy with centrotemporal spikes: a 6-month randomized, double-blind, placebo-controlled study. Sulthiame Study Group. AB - PURPOSE: To evaluate the efficacy and tolerability of sulthiame (STM) as monotherapy in children with benign childhood epilepsy with centrotemporal spikes (BECTS). METHODS: Sixty-six BECTS patients entered a 6-month double-blind trial and were randomized to receive either STM (5 mg/kg/day) or a placebo. All patients had had two or more seizures during the 6 months preceding the trial and were aged 3-11 years. Seizures were recorded by parents in a diary. STM plasma levels and electroencephalograms (EEGs) were overseen by patient-blinded observers. The primary effectiveness variable was the rate of treatment failure events (TFEs) per group. TFEs consisted of a first seizure after a 7-day run-in period, intolerable adverse events (AEs), development of another epileptic syndrome, or termination of the trial by parents or patient. RESULTS: Twenty-five of the 31 STM-treated patients (81%) and 10 of the 35 placebo-treated patients (29%) completed the trial without any TFEs (p = 0.00002). Most TFEs were seizures (n = 4 for the STM patients, n = 21 for the placebo group). Parents requested termination for two placebo-treated patients. Four patients were terminated for administrative reasons. No patient was withdrawn for AEs. While all patients displayed at least one specific focus in either the awake or asleep EEG initially, 11 STM-treated patients had a normal awake EEG and 10 had a normal asleep one after 6 months. CONCLUSIONS: STM was remarkably effective in preventing seizures in patients with BECTS. Patients suffering from > or = 2 seizures during the past 6 months had a high risk of early recidivism. STM was well tolerated and should be considered for children with BECTS who are in need of treatment. PMID- 11051124 TI - Gabapentin versus vigabatrin as first add-on for patients with partial seizures that failed to respond to monotherapy: a randomized, double-blind, dose titration study. GREAT Study Investigators Group. Gabapentin in Refractory Epilepsy Add-on Treatment. AB - PURPOSE: Our objective was to compare the efficacy and safety of gabapentin and vigabatrin as first-line add-on treatment in patients with partial epilepsy. METHODS: This was a multicenter, double-blind, randomized dose titration study. After baseline assessment and randomization, the dose could be increased if seizures persisted and reduced if side effects occurred. Health-related quality of life was assessed at baseline and at the end of the study. By a protocol amendment post hoc, all randomized patients were offered a standardized perimetry examination at the end of the study. Improvement rate was the proportion of patients with a reduction of seizure frequency of at least 50% during an 8-week period without any adverse events causing withdrawal. RESULTS: One hundred two patients were randomized and analyzed on an intent-to-treat basis. The improvement rate was 48% in the gabapentin group and 56% in the vigabatrin group. The improvement rate, when per protocol criteria were fulfilled, was 57% in the gabapentin group and 59% in the vigabatrin group. The proportion of seizure-free patients was 31% in the gabapentin group and 39% in the vigabatrin group. There was no difference in quality-of-life scores between the groups. Perimetry after termination of the study on 64 patients showed abnormal results in 3 of 32 patients in the vigabatrin group. CONCLUSION: Approximately one third of the patients in both groups became seizure-free. Although no major differences were seen in terms of the improvement rate between the groups, equivalence between the two drugs was not found. PMID- 11051125 TI - Epilepsy and antiepileptic drug therapy in juvenile neuronal ceroid lipofuscinosis. AB - PURPOSE: To survey the characteristics of epilepsy in patients with juvenile neuronal ceroid lipofuscinosis (JNCL) and determine the antiepileptic drug (AED) treatment most suitable for these patients. METHODS: The study included 60 patients with JNCL; their mean age was 16.5 years (range 5-33). The age at onset of epilepsy, type of seizures, effect of the first AED on seizures, and the current seizure frequency and AED therapy were studied. The side effects of the AEDs were also clarified. RESULTS: Fifty of the 60 patients had epilepsy. Patients' first epileptic seizure occurred at a mean age of 10.0 years (range 5 16), the most common type being generalized seizures. As the first AED tried, valproate (VPA) and lamotrigine (LTG) appeared equally effective, with 80% of the patients responding to these AEDs. During the study year, the median seizure frequency was four seizures a year (range 0-120), and 72% of the patients had good or satisfactory seizure control (0-6 seizures a year). In the different AED therapy groups, the proportion of patients with good or satisfactory seizure control ranged from 25% to 100%. LTG in monotherapy or in combination with clonazepam (CZP) was superior to other AEDs or combinations, but VPA also seemed effective. Adverse effects leading to the discontinuation of an AED were observed in 25% of the patients, most frequently in patients receiving phenobarbital (PB). No patient receiving LTG had to discontinue the drug due to adverse effects. CONCLUSION: Epilepsy in JNCL can usually be successfully treated with the current AEDs. In Finnish patients with JNCL, treatment is based on LTG, or, secondarily, VPA. In combination therapy, CZP seems a valuable add-on AED. PMID- 11051126 TI - Does neuroticism influence cognitive self-assessment after epilepsy surgery? AB - PURPOSE: To examine how cognitive, personality, and seizure outcome variables influence the subjective cognitive functioning of patients with refractory temporal lobe seizures after epilepsy surgery. METHODS: Thirty-three consecutive patients with drug-resistant partial epilepsy who underwent surgical treatment at a tertiary referral university epilepsy center were tested before surgery and 1 year after surgery. Objective cognitive and subjective cognitive functioning tests were used, and personality was assessed. Seizure control was operationalized as a dichotomous variable. RESULTS: A significant inverse relationship was found between neuroticism and subjective cognitive functioning. None of the other pre- and postoperative cognitive and surgery outcome variables were significant predictors of subjective cognitive functioning, even after controlling for the effect of neuroticism. CONCLUSIONS: Subjective and objective memory functioning are independent in patients with epilepsy after surgical treatment. Subjective memory functioning appears to be related not to seizure relief but to neuroticism. These data suggest that psychological factors such as personality traits predisposing to emotional distress should be taken into consideration in the clinical management and counseling of patients undergoing epilepsy surgery. PMID- 11051127 TI - The "forgotten" cross-tolerance between phenobarbital and primidone: it can prevent acute primidone-related toxicity. AB - PURPOSE: We report on the effect that pretreating patients with phenobarbital has on averting adverse events when primidone is introduced. METHODS: Thirty patients with intractable partial epilepsy were pretreated with phenobarbital before starting primidone. Therapy with primidone was started at a dosage of 500 mg/day, and the phenobarbital was stopped. The primidone dose was then increased by 125 to 250 mg every 3 weeks until adverse events or a seizure-free state was reached. All previous antiepileptic medications were tapered down to yield a primidone monotherapy regimen. RESULTS: Twenty-six patients (87%) tolerated the introduction of primidone with minimal or no adverse events. Only one patient had to discontinue primidone during the initial 4 weeks because of severe dizziness. This was the only patient in whom primidone monotherapy could not be reached because of adverse events. Three other patients experienced dizziness severe enough to interfere with their activities. This symptom disappeared in two patients after the dose was lowered; in the other patient, primidone was stopped and phenobarbital was restarted for another 4 days. No symptoms recurred when primidone was reintroduced on the fifth day. CONCLUSIONS: Pretreatment with phenobarbital can minimize the occurrence of intolerable adverse events associated with the introduction of primidone. PMID- 11051128 TI - Significance of cerebellar atrophy in intractable temporal lobe epilepsy: a quantitative MRI study. AB - PURPOSE: To determine the incidence of cerebellar atrophy (CA) in patients with intractable temporal lobe epilepsy, whether any clinical factors are significantly associated with CA, whether CA is unilateral or asymmetric and whether this feature has any relationship to the side of epileptogenicity, and whether the presence of CA is related to epilepsy surgery outcome. METHODS: We developed a magnetic resonance imaging method of measuring the presurgical volumes of the cerebellar hemispheres of 185 patients who underwent temporal lobectomy for intractable epilepsy and of 80 control subjects. In addition, cerebellar volumes were normalized to the total brain volumes. CA was determined as being present when the measured volume was smaller than two standard deviations from the mean value found in control subjects. RESULTS: Both absolute and normalized cerebellar volumes were found to be significantly reduced in the epilepsy patients compared with the control subjects. Without normalization of the cerebellar volumes, CA was present in 25.9% of the epilepsy patients; with normalization, it was present in only 16.2%. The atrophy was symmetric between the cerebellar hemispheres, and there was no significant difference in volume between the hemisphere ipsilateral and the hemisphere contralateral to the side of the temporal lobectomy. The duration of epilepsy was significantly longer and the age at onset of epilepsy was younger in patients with CA than in those without CA. The presence of CA was not associated with the outcome of temporal lobectomy. CONCLUSIONS: CA is symmetric and common in patients with intractable temporal lobe epilepsy. However, the results suggest that the atrophy in one third of patients with CA also proportionately affects the cerebral hemispheres. The duration of epilepsy and the age at onset of epilepsy are associated with the occurrence of CA. Seizure control after temporal lobectomy is not influenced by the presence of CA. PMID- 11051129 TI - Does the intracarotid amobarbital procedure predict global amnesia after temporal lobectomy? AB - PURPOSE: The intracarotid amobarbital procedure (IAP) is widely used to help predict who might be at risk for postoperative amnesia after unilateral temporal lobectomy for intractable seizures. We describe the memory outcome in 10 patients who underwent standard temporal lobectomy, including mesial temporal structures, despite failing the memory portion of the IAP after injections both ipsilateral and contralateral to the resected seizure focus. METHODS: Data for seven of the study subjects were obtained through a retrospective review of patients assessed on a surgical epilepsy unit during a 15-year period who failed the Montreal Neurological Institute IAP memory protocol after both ipsilateral and contralateral injections and subsequently underwent unilateral temporal lobectomy. More recently, we have studied temporal lobectomy patients who failed the Medical College of Georgia memory protocol after both ipsilateral and contralateral injections (n = 3). Preoperative and postoperative memory test scores were compared, and data regarding seizure outcome and self-perception of postoperative memory were collected. RESULTS: At follow-up, none of the patients presented with a pattern indicative of a global amnesia, and 80% demonstrated >90% improvement in their seizure disorder or were seizure-free. CONCLUSIONS: These findings indicate that bilateral memory failure on the IAP does not preclude the removal of an epileptogenic temporal lobe or a successful surgical outcome. In addition, the findings raise questions regarding the validity of the IAP and the possibility that memory may be reorganized in patients with a long history of temporal lobe epilepsy. PMID- 11051130 TI - Nonepileptic seizure outcome varies by type of spell and duration of illness. AB - PURPOSE: To determine whether differences in clinical manifestations of psychogenic nonepileptic events are associated with differences in outcome and whether the length of illness before diagnosis correlates with outcome. METHODS: We reviewed ictal videotapes and EEGs in 85 patients diagnosed with exclusively nonepileptic psychogenic seizures during inpatient CCTV-EEG monitoring at the University of Michigan between June 1994 and December 1996. They were classified into groups of similar ictal behaviors. Fifty-seven of these patients were available to respond to a follow-up telephone survey about their condition 2-4 years after discharge. We examined demographics, baseline EEG abnormalities, and outcome of treatment interventions. We also evaluated whether interventions were more likely to succeed if patients were diagnosed early in the course of the illness. RESULTS: We found that the largest groups consisted of patients with motionless unresponsiveness ("catatonic," n = 19) and asynchronous motor movements with impaired responsiveness ("thrashing," n = 19). Infrequent signs included tremor, automatisms, subjective events with amnesia, and intermittent behaviors. There was a higher incidence of baseline EEG abnormalities in the thrashing group (31%) than in the catatonic group (0%). There was a higher incidence of complete remission of spells in the catatonic group (53%) than in the thrashing group (21%). Patients who had a more recent onset of seizures (most often within 1 year) were much more likely to have remission of spells after diagnosis. CONCLUSIONS: Classification of nonepileptic seizures is useful in predicting outcome and may be valuable in further investigation of this complex set of disorders. PMID- 11051131 TI - Psychosocial impact of epileptic seizures in a Dutch epilepsy population: a comparative Washington Psychosocial Seizure Inventory study. AB - PURPOSE: The psychosocial functioning of epilepsy patients from the Netherlands was investigated and compared with results from other countries. The impact of epilepsy was also studied in two different groups of Dutch epilepsy patients, inpatients and outpatients. METHODS: The Washington Psychosocial Seizure Inventory (WPSI) was used to study the psychosocial problems of 134 Dutch outpatients and 181 Dutch inpatients. WPSI profiles were compared with those from the former German Democratic Republic (West Germany), Finland, Canada, the United States, Chile, and Japan. RESULTS: For the Dutch epilepsy patients, most of the psychosocial problems were experienced by inpatients; they had serious problems in emotional, interpersonal, and vocational adjustment, adjustment to seizures, and overall psychosocial functioning. Seizure-free outpatients, however, experienced significant problems only in the emotional adjustment area. Comparing the outcomes of various countries, Dutch outpatients and patients from West Germany and Finland experienced the least psychosocial difficulties, whereas epilepsy patients from Chile, Japan, and Canada have serious problems in most areas of psychosocial functioning. CONCLUSIONS: Patients with epilepsy experience psychosocial problems, although the amount of psychosocial difficulties depends on the seizure frequency and the culture that patients live in. PMID- 11051132 TI - The safety of rapid valproic acid infusion. AB - PURPOSE: The recommended rate of administration of valproic acid injection is 20 mg/min. Drug delivery at this rate may be inadequate for expeditious control of seizures. The safety of rapid infusion of valproic acid has not been established, and this study was designed to explore the effects of rapid infusion in patients with acute seizures. METHODS: Twenty patients with acute repetitive seizures received 20 mg/kg loading doses of valproic acid. Infusion rates ranged from 33.3 to 555 mg/min (median, 200 mg/min). Sixteen patients had received previous or concomitant antiepileptic drugs, with inadequate seizure control. Heart rate, blood pressure, and respiratory rate were measured before infusion and at frequent intervals for 1 hour after infusion. Patients were also observed for changes in level of alertness and signs of local irritation. RESULTS: No patient exhibited a decline in level of consciousness or respiratory function. Two patients with significant contributing factors exhibited declines in blood pressure requiring vasopressors. No significant local irritation was reported. Although efficacy was not a measured end point, seizures were abolished in all patients. CONCLUSIONS: Infusion of valproic acid at rates between 33 and 555 mg/min is well tolerated. No serious adverse effects attributable to the rapid infusion of valproic acid were encountered, although valproic acid, along with other factors, may have contributed to the hypotension in two patients. Intravenous valproic acid is an option for the control of acute seizures. PMID- 11051133 TI - Commission on Neurosurgery of the International League Against Epilepsy (ILAE) 1993-1997: recommended standards. PMID- 11051134 TI - Commission on Diagnostic Strategies: recommendations for functional neuroimaging of persons with epilepsy. PMID- 11051135 TI - Dermatopharmacology of ciclopirox nail lacquer topical solution 8% in the treatment of onychomycosis. AB - Ciclopirox is a synthetic hydroxypyridone antifungal agent. In contrast to the azoles, glucuronidation is the main metabolic pathway of ciclopirox; therefore interactions with drugs metabolized via the cytochrome P450 system are unlikely Ciclopirox is also distinct from the common systemic agents, which interfere with sterol biosynthesis. In fact, ciclopirox chelates trivalent cations (such as Fe3+), inhibits metal-dependent enzymes that are responsible for degradation of toxic metabolites in the fungal cells, and targets diverse metabolic (eg, respiratory) and energy producing processes in microbial cells. Ciclopirox is a broad spectrum antimicrobial with activity against all the usual dermatophytes as well as yeast and nondermatophyte molds. It has demonstrated activity against gram positive and negative bacteria, including resistant strains of Staphlococcus aureus. Ciclopirox exhibits fungal inhibitory activity (minimum inhibitory concentration < 4 microg/mL for dermatophytes) as well as fungicidal activity; to date resistance to the drug has not been identified. Ciclopirox has been formulated in a nail lacquer delivery system. After evaporation of volatile solvents in the lacquer, the concentration of ciclopirox in the remaining lacquer film reaches approximately 35%, providing a high concentration gradient for penetration into the nail. Radiolabel data demonstrate penetration into infected nails after only 1 application of the lacquer. Ciclopirox nail lacquer is a topical product that provides an active fungicidal agent in a delivery system capable of promoting nail penetration. With repeated applications, the antifungal agent is homogeneously distributed through all layers of the toenail achieving concentrations of ciclopirox in excess of inhibitory and fungicidal concentrations for most pathogens. Although ciclopirox readily penetrates nails, very low levels of ciclopirox are recoverable systemically, even after chronic use. Ciclopirox nail lacquer 8% is a topical product that provides an active fungicidal agent in a delivery system capable of penetrating nails. PMID- 11051136 TI - Ciclopirox nail lacquer topical solution 8% in the treatment of toenail onychomycosis. AB - BACKGROUND: Onychomycosis is a relatively common condition affecting toenails more than fingernails. It is caused predominantly by dermatophytes. Onychomycosis can cause pain and discomfort and has the potential to be a source of morbidity. OBJECTIVE: We evaluated the efficacy and safety of ciclopirox nail lacquer solution 8% used to treat onychomycosis of the toe in the United States and in centers worldwide. METHODS: Two identically designed, double-blind, vehicle controlled, parallel group multicenter studies were performed in the United States to evaluate the use of ciclopirox nail lacquer to treat mild to moderate toe onychomycosis caused by dermatophytes. In the first study, 223 patients were randomized to treatment (ciclopirox group: 112, vehicle group: 111), and in the second study, 237 subjects were randomized (ciclopirox group: 119, vehicle group: 118). Before randomization, patients were to have clinical features of onychomycosis in at least one great toe with positive light microscopic examination and a positive dermatophyte culture. The test material was applied daily for a period of 48 weeks to all toenails and affected fingernails, covering the entire nail plate and approximately 5 mm of surrounding skin. At baseline, subjects had between 20% to 65% area of target nail involved. Physician's assessments were carried out every 4 weeks, and mycologic evaluation and photographic planimetry using standardized photographs were performed every 12 weeks during the 48 weeks of treatment. In studies conducted outside the United States, patients were also to have clinical, microscopic, and culture evidence of onychomycosis. However, these studies included some patients infected with nondermatophyte organisms (eg, Candida species), and the area of nail involvement was generally greater than observed in the US studies. Treatment regimens also varied in the non-US studies with lacquer applications that were sometimes less frequent than the once daily treatment used in the US studies (eg, alternate day or twice weekly). In addition, the typical duration of treatment was 6 months in the non-US studies as compared with 48 weeks in the United States. Outcome measures were similar to those used in the US trials, although a non-photographic planimetric method was used to quantify disease extent. RESULTS: Data from the pivotal US trials have demonstrated that ciclopirox nail lacquer 8% topical solution is significantly more effective than placebo in the treatment of onychomycosis caused by Trichophyton rubrum, and of mild to moderate toe onychomycosis without lunula involvement. At the end of the 48-week treatment period, the mycologic cure rate (negative culture and negative light microscopy) in study I was 29% vs 11% in the ciclopirox and vehicle groups, respectively. Similarly, the mycologic cure rate for study II was 36% vs 9%, respectively. In the non-US studies, the mycologic cure rates ranged from 46.7% to 85.7%. In addition, ciclopirox nail lacquer has demonstrated a broad spectrum of activity with efficacy against Candida species and some nondermatophytes in non-US studies. Ciclopirox nail lacquer is considered extremely safe regarding causally related treatment emergent adverse-effects (TEAEs), with most TFAEs transient and localized to the site of action (eg, erythema and application site reaction). In the US studies, TFAEs were generally mild and cleared while the patient continued to use the nail lacquer. CONCLUSIONS: Studies conducted worldwide demonstrate the efficacy of ciclopirox nail lacquer for the treatment of finger and toe onychomycosis. Both controlled and open-label studies confirm the excellent safety profile of this topical therapy. Thus, the nail lacquer provides a treatment choice with a favorable benefit-to-risk ratio. With its novel mechanism of action and its topical route of administration, ciclopirox nail lacquer offers an innovative approach to the treatment of this often difficult-to-manage disease PMID- 11051137 TI - Pharmacoeconomic analysis of ciclopirox nail lacquer solution 8% and the new oral antifungal agents used to treat dermatophyte toe onychomycosis in the United States. AB - BACKGROUND: Recently a novel topical nail lacquer, ciclopirox solution 8%, has been approved for the treatment of onychomycosis. OBJECTIVE: This was undertaken to determine the most cost-effective treatment for the treatment of dermatophyte onychomycosis of the toes in the United States in 2000. METHODS: The nature of the problem was defined. The drug comparators were ciclopirox nail lacquer, terbinafine, itraconazole (pulse), itraconazole (continuous), fluconazole, and griseofulvin. A decision analytic model that reflected the manner in which pedal tinea unguium is managed was produced. Studies that have evaluated the efficacy of the nail lacquer and the oral antifungal agents for this indication were identified. Appropriate studies were used in a meta-analysis to determine the mycologic and clinical response rates when the drug comparators are used for the treatment for toe dermatophyte onychomycosis. For each drug comparator a cost of regimen analysis was carried out. This is the sum of the drug acquisition cost, the cost of medical management, and the cost of managing adverse effects. Next, the expected cost of management was calculated, disease free days were determined, and a sensitivity analysis was conducted. RESULTS: For each comparator the meta-analytic average mycologic cure (MC) rate and clinical response (CR) rates were: ciclopirox nail lacquer (MC: 52.6 +/- 4.2%, CR: 52.4 +/ 9.0%), griseofulvin (MC: 41.1 +/- 20.4%, CR: 33.7 +/- 14.1%), itraconazole (continuous) (MC: 66.3 +/- 4.2%, CR: 70.3 +/- 4.2%), itraconazole (pulse) (MC: 70.8 +/- 5.7%, CR: 73.6 +/- 4.6%), terbinafine (MC: 77.2 +/- 4.0%, CR: 75.3 +/- 2.9%), and fluconazole (MC: 65.6 +/- 7.1%, CR: 66.5 +/- 11.7%). The cost of regimen for the drug comparators was: ciclopirox nail lacquer $325.2, griseofulvin $1413.1, itraconazole (continuous) $1410.2, itraconazole (pulse) $811.7, terbinafine $890.1, and fluconazole $966.8. The cost/mycologic cure rate and expected cost/expected symptom free day were, ciclopirox nail lacquer ($618.2, 1.69), griseofulvin $3438.2, 5.3), itraconazole (continuous) ($2126.9, 3.52), itraconazole (pulse) ($1146.4, 2.01), terbinafine ($1153.0, 2.14), and fluconazole ($1473.7, 2.10). The relative cost-effectiveness was ciclopirox nail lacquer 1.00, itraconazole (pulse) 1.19, fluconazole 1.24, terbinafine 1.27, itraconazole (continuous) 2.08, and griseofulvin 3.13. Sensitivity analysis indicated that ciclopirox nail lacquer was a cost effective alternative compared with the oral regimens of terbinafine, itraconazole (continuous), and griseofulvin when clinical response rate was used as the primary efficacy parameter. CONCLUSION: Ciclopirox nail lacquer solution 8% is a recent addition to the armamentarium of therapies available to the physician and patient for the treatment of onychomycosis. The nail lacquer is a cost effective agent compared with the oral antifungal therapies, terbinafine, itraconazole, fluconazole, and griseofulvin. PMID- 11051138 TI - Ciclopirox nail lacquer solution 8% in the 21st century. AB - Ciclopirox nail lacquer solution 8% has been shown to be effective in the treatment of dermatophyte onychomycosis of mild to moderate severity Other studies report the effectiveness of ciclopirox nail lacquer in onychomycosis caused by Candida sp and nondermatophyte molds. Ciclopirox nail lacquer may also be valuable in the treatment of early cases of reinfection/relapse. Ciclopirox nail lacquer solution 8% may be an important adjunct to oral antifungal therapy in certain presentations that might be poorly responsive to oral antifungal therapy alone (eg, lateral onychomycosis, longitudinal spike, dermatophytoma, and extensive onycholysis). In some cases, surgical therapies may need to be considered in addition to, or in preference to, topical nail lacquer treatment. The use of ciclopirox nail lacquer solution 8% as an adjunct to oral antifungal therapy may widen the spectrum of activity of the combination because of the broad spectrum of coverage provided by the lacquer. The use of combination therapy may be synergistic in terms of efficacy, enabling a reduction in the duration and cumulative dosage of oral therapy. This could result in a decrease in the frequency and severity of systemic adverse effects associated with the oral antimycotics and the need to be vigilant about drug interactions. Studies need to be conducted to determine the place of combination oral and topical lacquer therapy in the management of onychomycosis. PMID- 11051139 TI - Quantification of recirculation by thermodilution during venovenous extracorporeal membrane oxygenation. AB - PURPOSE: The aim of this study was to determine whether recirculation could be quantified by a thermodilution technique during venovenous (VV) extracorporeal membrane oxygenation (ECMO) in a rabbit model. METHODS: Five New Zealand white rabbits, mean weight, 4.5 (range, 3.7 to 5.7) kg, were anesthetized, instrumented, cannulated with a double-lumen catheter, and placed on VV ECMO. Serial injections of ice-cold saline were performed at the arterial arm of the circuit, and the resultant temperature change at various pump flows was measured at the venous arm of the circuit using a thermistor-tipped catheter and a cardiac output computer. Results were compared with the respective 100% recirculation measured with all the circuit flow passing through the bridge. RESULTS: Using linear regression, recirculation percentage could be calculated as: 19 + 0.1 x pump flow (R2 = 0.81, P < .005). Recirculation correlated positively with pump flow. Variability between results at each flow was less than 10%. CONCLUSIONS: Recirculation can be quantified during VV ECMO by measuring the change in temperature in the venous arm using a cardiac output computer after injection of a known quantity of ice-cold saline in the arterial side of the circuit. The effect of interventions to reduce recirculation can be assessed conveniently and reliably. PMID- 11051140 TI - Clinical implications of minimal disease in the bone marrow and peripheral blood in neuroblastoma. AB - PURPOSE: In patients with neuroblastoma (NB), minimal disease (MD) in bone marrow (BM) and peripheral blood (PB) is thought to play an important role in metastasis. The current study was designed to evaluate the clinical implications of the detection of MD in NB at the initial diagnosis. METHODS: Expression of the neuroendocrine protein gene product 9.5 (PGP9.5) and tyrosine hydroxylase (TH) mRNA in BM and PB obtained from 18 patients with NB was investigated by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: MD was detected in the BM obtained from 4 of 14 localized NB patients and also in the PB from 2. However, it was found also in both the BM and PB obtained from all 4 patients with metastatic NB. Two of the 4 MD-positive patients with localized NB had metastatic recurrence after a complete tumor excision. They also had unfavorable biological prognostic factors compared with the other 2 who did not have recurrent disease. CONCLUSION: MD detected by RT-PCR in the BM and the PB of patients with NB thus suggests a risk for metastatic disease, which in association with other unfavorable biological features may indicate a poor prognosis. PMID- 11051141 TI - Defective fibroblast growth factor signaling allows for nonbranching growth of the respiratory-derived fistula tract in esophageal atresia with tracheoesophageal fistula. AB - BACKGROUND/PURPOSE: The fistula tract in esophageal atresia with tracheoesophageal fistula (EA-TEF) appears to arise from a trifurcation of the embryonic lung bud. Subsequently, it does not branch like the other bronchi, which also arise from the lung bud. Previous results have implied that aberrant mesenchymal-epithelial signaling in the developing foregut, possibly involving fibroblast growth factors, may allow for the nonbranching growth of the fistula, and the ultimate development of the fistula tract in TEF. METHODS: Adriamycin injections into pregnant rat dams induced EA-TEF formation in rat embryos. Control and Adriamycin-exposed embryos were harvested on the 13th gestational day, and the developing foregut was isolated with microdissection. mRNA was isolated from the developing fistula tract, embryonic lung, and normal embryonic esophagus. Reverse transcription-polymerase chain reaction (RT-PCR) for the IIIb splice variant of the FGF2R receptor was performed. Foregut specimens also were processed for histologic analysis, and immunofluorescence for FGF1 was performed. RESULTS: FGF2R-IIIb is specifically absent from the developing fistula tract in TEF, whereas it is present in the normal developing lung and esophagus. FGF1 also is uniquely absent from the developing fistula tract, but it is present in the normal lung mesenchyme. CONCLUSIONS: FGF1, FGF7, and FGF10 are critical mesenchymal factors that mediate proliferation and branching morphogenesis by the developing respiratory epithelium. The absence of FGF2R-IIIb, the obligate common receptor for FGF7 and FGF10, from the fistula tract, and the absence of FGF1 in the fistula tract mesenchyme, collectively imply the absence of a specific FGF signaling pathway in the developing fistula tract. This absence of FGF signaling could explain the lack of branching by the developing fistula tract as it grows caudally in the abnormally developing embryo. Downregulation of these components of the FGF signaling pathways may allow for a patterned compensation by the embryo for the proximal foregut atresia in this anomaly. This compensation may then reestablish gastrointestinal continuity as the fistula tract connects to the developing stomach. PMID- 11051142 TI - Needlescopic surgery for cryptorchidism: the initial series. AB - PURPOSE: The aim of this study is to report the initial experience with needlescopic surgery (2-mm optics and instrumentation exclusively) for the cryptorchid testicle. METHODS: Ten patients (age 8 months to 37 years) underwent 12 needlescopic procedures: orchiopexy (n = 8), orchiectomy (n = 2), and diagnostic exploration with attempted excision of testicular remnant (n = 2). Two patients underwent bilateral needlescopic orchiopexy. Needlescopic (2 mm) optics and instrumentation were used exclusively in the pediatric patients. RESULTS: All procedures were completed successfully by needlescopic techniques. Mean surgical time was 110 minutes (range, 60 to 180 minutes), and blood loss was 6 mL (range, 0 to 20 mL). There were no intraoperative complications. All procedures were performed on an outpatient basis. In all 8 orchidopexies, the testis was successfully brought to a scrotal position. CONCLUSIONS: Needlescopic techniques allow safe performance of various procedures for a cryptorchid testicle. The cosmetic result is excellent. PMID- 11051143 TI - Bone setter's gangrene. AB - BACKGROUND/PURPOSE: Little published data exist on the morbidity and mortality associated with poor trauma care in developing countries. This report highlights our experience with iatrogenic limb gangrene related to fracture management by traditional bone setters. METHODS: Children with "bone setter's" gangrene were identified from a prospectively recorded pediatric surgery database at the Royal Victoria Hospital in Banjul, The Gambia--the main government referral hospital. RESULTS: Nine children were treated for bone setter's gangrene during a 29-month period. The average age was 8.2 years (range, 5 to 14 years). Bone setter's gangrene was more common in boys (male to female ratio, 2:1) and occurred almost exclusively (89%) in children from rural areas where access to health care was limited. Five children (56%) underwent proximal extremity amputations. Complications included 1 death related to sepsis, a case of chronic osteomyelitis at an amputation site, and a contracted insensate hand in a child whose parents refused amputation. CONCLUSIONS: Bonesetter's gangrene is a preventable complication that results from a failure of child health planners to recognize the importance of basic trauma care. Management of fractures should be considered an essential component of child health programs in developing countries. PMID- 11051144 TI - One-stage Duhamel-Martin procedure for Hirschsprung's disease: a 5-year follow-up study. AB - BACKGROUND/PURPOSE: With the introduction of the Endo-GIA stapling device the 1 stage Duhamel-Martin procedure became feasible for neonates and infants. Early results were promising. So far there were no meaningful data on the long-term functional results. This study shows the 5-year follow-up results after 1-stage Duhamel-Martin procedure for Hirschsprung's disease in neonates and infants. The results are compared with a historical group of patients from the same institution undergoing a 3-stage procedure. METHODS: Between September 1991 and December 1993 Hirschsprung's disease was diagnosed in 29 children. In 22 of them the disease was found within the first 2 months of life. In 19 children aganglionosis was restricted to the rectosigmoid colon. In 10 the innervation disturbance extended further, twice with involvement of the distal ileum. Initial treatment consisted of daily rectal irrigation. Postoperative follow-up on a regular out-clinic basis was 6 years (range, 5 to 7 years). Patients were scored for fecal continence, soiling, the use of laxatives, cannulae or rectal irrigation, enterocolitis, gain of body weight, and length. RESULTS: There were no intraoperative complications. The median postoperative stay was 7.7 days. Seven children encountered complications for which admission was necessary. Ultimately, 15 children have normal spontaneous defecation. Eight children display irregular soiling, without using laxatives. At 5-year follow-up 6 children are still on some sort of laxative or rectal irrigation. Mean growth and body weight is along the P50 and P50 to 90, respectively. These functional results are no different from those in the patients after 3-stage Duhamel-Martin procedure. CONCLUSIONS: There appears to be no difference in functional outcome after 1- or multiple-stage Duhamel-Martin procedure for Hirschsprung's disease after 5 to 7 years. The majority of children seem to fare well with restrictive need of laxatives. The advantage of a 1-stage procedure is the prevention of stoma-related complications, 1 or 2 additional operations, and extra scar formation. PMID- 11051145 TI - Unsatisfactory experience with 'minimal intervention management' for gastroschisis. AB - PURPOSE: The authors report, for cautionary reasons, their trial with "minimal intervention management" for gastroschisis. After the successful innovative experience with this approach, which Bianchi and Dickson described, they utilized it in 4 consecutive patients. METHODS: In the delivery room a plastic bag was placed over the intestines, which rested in a dependent position to reduce edema. The stomach was decompressed and the patients kept warm. Intravenous fluid at a maintenance rate was given. After about 5 hours an attempt at closure was undertaken in the newborn intensive care unit without anesthesia. An assistant lifted the anterior abdominal wall by applying upward traction on the umbilical cord. Over about 25 minutes the intestines were placed in the coelom, which was closed with a single suture. RESULTS: The outcome was uncomplicated in the first of 4 consecutive patients. The second patient had abdominal compartment syndrome requiring a silo and subsequent resection and has chronic malabsorption 16 months later. The third had an enterocutaneous fistula at 5 weeks that required a small bowel resection. Bedside closure was abandoned in the final case because too much resistance was encountered. She underwent primary repair in the operating room and eventually died of sepsis with intestinal dysmotility. CONCLUSIONS: The "minimal intervention approach" can be effective in some patients who have gastroschisis. This experience suggests that selection criteria are needed before this method can be recommended. PMID- 11051146 TI - Physiotherapy as an adjuvant to the surgical treatment of anterior chest wall deformities: a necessity? A prospective descriptive study in 21 patients. AB - PURPOSE: The authors postulated that physiotherapy as an adjuvant to the surgical treatment of anterior chest wall deformities is only indicated if specific abnormalities can be found that could be corrected by physiotherapy. The purpose of this study is to investigate whether such abnormalities can be found and to evaluate their course during a postoperative period of 18 months. METHODS: Twenty one patients, 16 with pectus excavatum and 5 with pectus carinatum, were evaluated 6 weeks before and 6 weeks, 6 months, and 18 months after surgical correction. Postural impairments, spinal mobility and curvature, muscle strength, and muscle length were evaluated. RESULTS: Preoperatively, poor posture was seen in 10 patients, nonstructural scoliosis in 11, and abdominal muscle weakness in 4 patients. None of the patients had restriction of spinal mobility or shortened pectoral muscles. Six weeks after surgery, poor posture was seen in 9, nonstructural scoliosis in 11, and abdominal muscle weakness in 10 patients. The authors found a higher percentage of recovery for abdominal muscle weakness than for poor posture (90% versus 33%, respectively). CONCLUSIONS: The authors found preoperative postural impairments in 52% of their patients, in patients with pectus carinatum as well as in patients with pectus excavatum. In patients without postural impairments, physiotherapy is not necessary, with the exception of postoperative pulmonary care. PMID- 11051147 TI - The role of melatonin in prevention of intestinal ischemia-reperfusion injury in rats. AB - BACKGROUND/PURPOSE: The aim of this study was to determine the effect of melatonin, a hormone that is known as an antioxidant, on the prevention of tissue damage during mesenteric ischemia/reperfusion (I/R). METHODS: A total of 40 young Wistar-albino rats were divided equally into 4 groups with varied treatment. Group 1 was control (sham), group 2 was I/R, group 3 was I/R plus melatonin (10 mg/kg) and group 4 was I/R plus melatonin (20 mg/kg). I/R was realized as follows: after laparatomy, a microvascular atraumatic clip was placed across the superior mesenteric artery (SMA) under general anaesthesia, and it was removed after ischemia for 30 minutes. The first dose of melatonin was applied intraperitoneally at the start of reperfusion. The second and third doses were applied intramuscularly on the first and second day. Only SMA dissection under general anaesthesia was carried out in the control group rats. On the third day of the study all the rats were killed, and their bowels were removed. Malondialdehyde (MDA) levels were assayed as an index of lipid peroxidation reflecting free radical reaction in the intestine. Histopathologic analysis was made using light microscopy in a blind fashion. RESULTS: The levels of tissue MDA were found to be significantly lower in groups 3 and 4 compared with group 2 (P < .05). The MDA levels of group 4 did not differ significantly from that of the control group (P > .05). The histopathologic results were consistent with the MDA levels. CONCLUSION: These results suggest that melatonin has a strong antioxidant effect in preventing intestinal I/R damage, and that this effect is exerted in a dose-dependent manner. PMID- 11051149 TI - Treatment of proximal hypospadias with a tubularized island flap urethroplasty and the onlay technique: a comparative study. AB - BACKGROUND/PURPOSE: The onlay island flap urethroplasty was first described in the repair of mid and distal penile hypospadias. Since then, this technique has been increasingly used in more severe cases of hypospadias, because of the complications of tubularized flaps, mainly megaurethra and proximal anastomotic strictures. The aim of this study was to compare the morbidity of these 2 techniques. METHODS: Between April 1994 and December 1998, 80 patients underwent surgical treatment for hypospadias. A tubularized island flap (Ducketttechnique) was performed in 42 cases, and the onlay island flap technique was used in 38 patients. The authors retrospectively compared the complication rate and type of these 2 procedures. RESULTS: Altogether, fistula was the most frequent complication without any significant difference between the 2 groups (21.4% for Duckett technique and 18.4% for onlay repair; P > .05). However, the anastomotic stricture was much more common in the tubularized flap group (7.14% v 2.63%; P < .05). Moreover, a megaurethra was found only in the Duckett technique group (4.7%). There was no case of chordee recurrence, but 6 patients (15.7%) treated with the onlay technique required urethrolysis including dissection of the chord behind the urethral plate, and in the other 3 patients of the same group (7.9%), a dorsal Nesbit plication also was necessary. In all these cases, the urethroplasty included an island cutaneous flap to provide ventral coverage to the neourethra. CONCLUSIONS: The authors conclude that both techniques present similar complications. However, proximal strictures and megaurethra are more common after the Duckett technique. This procedure is of choice in patients with scrotal hypospadias. Conversely, the onlay repair should be completed with other procedures (urethrolysis, dorsal Nesbit plication) to obtain good results in patients with severe degree of chordee. PMID- 11051148 TI - Heart-related indices in experimental diaphragmatic hernia. AB - BACKGROUND: Heart-related indices have been suggested as useful tools to evaluate left ventricular (LV) hypoplasia, which might predict the outcome of fetuses and infants with congenital diaphragmatic hernia (CDH). The current study analyzed the behavior of such indices in the nitrofen-induced CDH rat model. METHODS: Dated pregnant Wistar rats received at day 9.5 of gestation either a dose of 100 mg of nitrofen or just the vehicle. Body, lung, and heart weights were measured in 12 newborn rats not exposed to nitrofen (Ctrl group) and 68 animals exposed to nitrofen: 30 without CDH (non-CDH group) and 38 with left CDH (CDH group). Each heart was fragmented in 7-microm thick sections. Only hearts with no evidence of cardiac morphologic defects (CMD) were studied further to estimate right and left ventricular cavity volumes, septal, right, and left ventricular free wall masses. These parameters allowed the calculation of the cardio-ventricular (CVindex) and LV mass indices. The aorta-to-pulmonary artery ratio also was calculated. RESULTS: Excluding fetuses with CMD, the heart-to-body weight ratio was reduced significantly in animals exposed to nitrofen, whereas no significant differences were observed between non-CDH versus CDH groups. Although the left and right ventricular cavity volumes were both reduced significantly in nitrofen-treated rats, they were not changed significantly by the existence of CDH, and the calculated CVindex was similar in the 3 groups. Estimated septal and LV masses were reduced markedly in the nitrofen-treated animals and further reduced by the presence of CDH. However, when LV mass was normalized (LV mass index) the difference became restricted to the animals exposed to nitrofen but was not influenced by the presence of CDH. Finally, the aorta-to-pulmonary artery ratio was similar in all studied groups. CONCLUSIONS: The results of the current study suggest that, although nitrofen had been responsible by global heart hypoplasia, the presence of CDH was not associated with significant underdevelopment of the heart or of the LV in rat fetuses without CMD. Based on these results, we think that the evidence for prenatal counseling based on heart-related indices should be critically reconsidered. PMID- 11051150 TI - The anterior mediastinal approach for management of tracheomalacia. AB - BACKGROUND: Tracheomalacia occurs as a primary developmental defect or may be secondary to vascular compression. It is common in association with esophageal atresia and tracheoesophageal fistula. Collapse of the weak trachea is a cause of recurrent respiratory symptoms but may be severe and life threatening. METHODS: Between 1978 and 1999 at Sheffield Children's Hospital and The Royal Manchester Children's Hospital, of 16 children with clinically significant symptoms of tracheomalacia 8 underwent combined aortopexy and tracheopexy, 1 had aortopexy alone, 4 only had a tracheopexy, and 3 had tracheal reinforcement with free costal cartilage ring grafts. The surgical approach was limited to a low cervical skin crease incision with a midline manubrial split providing extrapleural access to the anterior mediastinum and allowing for all surgery under direct unimpaired vision. RESULTS: Ten children did not require postoperative ventilatory support. Four underwent ventilation for a few hours or days. One child required CPAP for 4 months for residual tracheomalacia and a further child, who had 3 operations to insert 11 costal cartilage ring grafts, underwent ventilation intermittently for 6 months. Adequate tracheal patency could be verified by intraoperative tracheoscopy and was sustained postoperatively so that only 1 child with associated bilateral vocal cord paralysis came to tracheostomy. Four children required prolonged hospitalization because of residual tracheomalacia, 2 for bronchomalacia and 2 because of esophageal narrowing leading to further surgery. All other children were fit for discharge within 10 to 30 days of surgery. Long term follow-up has confirmed sustained tracheal improvement and resolution of the life-threatening features of tracheomalacia. CONCLUSIONS: The authors recommend the low skin crease transmanubrial approach, as described by Vaishnav and MacKinnon, for tracheopexy, aortopexy and for tracheal reconstruction for tracheomalacia. The approach gives excellent access for surgery under direct vision through a relatively avascular plane. It is associated with less morbidity than a conventional thoracotomy and leaves a more acceptable aesthetic scar. PMID- 11051151 TI - Inferior vena cava and right atrial thrombosis in children with nephroblastoma: diagnostic and therapeutic problems. AB - BACKGROUND: The neoplastic thrombus in Wilms' tumor rarely can extend in to the inferior vena cava or to the right atrium. The neoplastic thrombus usually is diagnosed concurrently with the tumor, although in some cases the diagnosis of the thrombus may precede the diagnosis of nephroblastoma. METHODS: Among 90 children with Wilms' tumor who were treated in the authors' unit, 4 had extensive tumor thrombosis of the inferior vena cava or the right atrium. One of these patients was found with a life-threatening thrombosis of the inferior vena cava and the right atrium, which was treated surgically; in this case, the diagnosis of nephroblastoma was made postoperatively. As for the 3 remaining patients the diagnosis of neoplastic thrombosis and Wilms' tumor was made simultaneously. RESULTS: In the first case, the patient underwent surgical excision of the thrombus with cardiopulmonary bypass and a short period of hypothermic cardiopulmonary arrest. In the other 3 cases the thrombus resolved with chemotherapy only. CONCLUSIONS: Surgical excision of extensive neoplastic thrombosis is suggested in the case of life-threatening thrombosis even with cardiopulmonary bypass. Chemotherapy is suggested in cases lacking clinical symptoms of thrombosis. PMID- 11051152 TI - Rectosigmoid pHi monitoring during experimental necrotizing enterocolitis. AB - BACKGROUND/PURPOSE: Gastric or rectosigmoid intramural pH (pHi) is considered a reliable indicator of splanchnic perfusion. The aim of this study was to evaluate whether rectosigmoid pHi reflects the severity of bowel damage in experimental necrotizing enterocolitis (NEC). METHODS: A total of 36 neonatal piglets, (median age, 3; range, 1 to 11 days; median weight, 2.5; range, 1.2 to 3.8 kg), were anesthetized, ventilated mechanically, and had invasive monitoring of hemodynamics. A sigmoid tonometer was inserted into the rectosigmoid colon. Enterocolitis was induced in 27 piglets by intraluminal injection of casein-d gluconate (16.0 mL/kg) into terminal 100 cm of the ileum. Nine control piglets received an equal amount of intraluminal saline. NEC was graded macroscopically as follows: 0, no changes; 1, mild; 2, moderate; and 3, severe. Histology was evaluated according to Chiu scale from 0 to 5. RESULTS: The macroscopical bowel injury in caseine-injected piglets was as follows: grade 3 (n = 6), grade 2 (n = 9), grade 1 (n = 12). All control piglets showed macroscopically normal bowel (grade 0). All affected bowels showed histologic changes (Chiu's scale 2 to 4). All study animals had an initial drop of pHi after injection of casein or saline. In control piglets and those with mild NEC (grade 1) pHi tended to return to preinjection level. In animals with moderate or severe NEC (grade 2 to 3) the initial drop was deeper and the pHi continued to decrease significantly throughout the experiment (P < .05). In the arterial pH and mean blood pressure there were no statistically significant differences between piglets with no NEC and mild NEC, and these with moderate or severe NEC. CONCLUSION: Drop in rectosigmoid pHi was the most sensitive and earliest sign of severe mucosal necrosis of ileum in this experimental NEC model. PMID- 11051153 TI - Antenatal dexamethasone enhances surfactant protein synthesis in the hypoplastic lung of nitrofen-induced diaphragmatic hernia in rats. AB - BACKGROUND/PURPOSE: Pulmonary hypoplasia is one of the main causes for the high mortality rate in patients with congenital diaphragmatic hernia (CDH). The expression of surfactant protein A in the hypoplastic CDH lung is reduced, and its concentration is decreased in the amniotic fluid of pregnancies complicated by CDH. In a CDH experimental model, prenatal glucocorticoid treatment has proved its efficacy in correcting the parameters of pulmonary biochemical and morphologic immaturity. The aim of this study was to investigate whether maternal administration of dexamethasone has any effect on the expression of surfactant protein A and surfactant protein B in nitrofen-induced experimental CDH rat model. METHODS: CDH was induced in pregnant rats after administration of 100 mg of nitrofen on day 9.5 of gestation (term, 22 days). Dexamethasone (Dex, 0.25 mg/kg) was given by intraperitoneal injection on days 18.5 and 19.5 of gestation. Cesarean section was performed on day 21 of gestation. The fetuses were divided into 3 groups: group I, control (n = 16); group II, nitrofen-induced CDH (n = 16); group III, nitrofen-induced CDH with antenatal Dex treatment (n = 16). Indirect immunohistochemistry was performed using alkaline-phosphatase-coagulated streptavidin using anti-SP-A and anti-SP-B polyclonal antibodies. Reverse transcription polymerase chain reaction (RT-PCR) was performed to evaluate relative amount of SP-A and SP-B mRNA expression. RESULTS: In the CDH lung (group II) we observed a markedly reduced number of type II pneumocytes positive for SP A, and SP-B was increased to a level close to that of the control group. The relative amount of SP-A and SP-B was reduced significantly in group II compared with controls (P < .05) and significantly increased in group III compared with group II animals (P < .01). CONCLUSION: These results suggest that antenatal glucocorticoid treatment increases the production of surfactant proteins in the CDH hypoplastic lung. PMID- 11051154 TI - Phagocytic reaction in contact with macroplastic: application in pediatric surgery. AB - BACKGROUND/PURPOSE: With regard to the problems of using polytetrafluoroethylene (PTFE) in the treatment of vesicoureteric reflux in children, the authors have tested polydimethylsiloxane (PDMS), which is conveyed by polyvinylpyrrolidone (PVP), a hydrogel. METHODS: The phagocytic reaction was tested with both PDMS and PVP. The phagocytic reaction of PDMS and PVP was tested in vitro, then the outcome of PVP was tested in vivo in the mouse. RESULTS: In vitro PVP was phagocytosed by mouse peritoneal macrophages. However, PDMS particles were not phagocytosed because of their large size. In vivo, PVP migrated to other organs but did not induce clear histologic lesions. CONCLUSIONS: Silicone (PDMS) has been used in the treatment of vesicoureteric reflux in children. It is now necessary to do tests on heavier animals with a long incubation period to know whether these particles migrate and what the histologic and clinical consequences might be. PMID- 11051155 TI - Epidemiologic aspects of pediatric multiple trauma in a Spanish urban population. AB - BACKGROUND/PURPOSE: Trauma is still the most frequent cause of mortality and disability in childhood and adolescence. An epidemiologic prospective study on children under 16 years of age with multiple trauma (MT) was conducted in a large Spanish urban university hospital over a 6-year period. METHODS: Of 1,937 children admitted at the pediatric trauma unit for musculoskeletal injuries from March 1988 until March 1994, 56 patients including 37 boys and 19 girls had MT. MT was considered when at least 2 long bones were fractured or there was a fracture of 1 long bone combined with an injury of 4 other major anatomic regions (face and neck, thorax, abdomen, or neural system). The severity of injuries was evaluated according to the modified injury severity score (MISS). RESULTS: Injury to pedestrians was the most frequent cause of MT (54%). The overall mortality rate of the series, including those children dying during transport to the hospital was 11.5%. The average MISS for the whole group was 15 (range, 5 to 59). Head trauma was the most frequent associated injury (52%), two thirds of which were considered minor injuries (Glasgow Coma Scale >15). Seventy-seven fractures were registered, 10% of which were open fractures. External fixation was the most common surgical technique among operated fractures. The average hospitalization period was 16 (median, 13; range, 1 to 150) days. Children with a MISS above 18 points showed a significant longer hospitalization period (mean, 31 +/- 45 days) as compared with those with MISS below 18 points (mean, 10 +/- 7 days; P < .05). There was a strong correlation between the MISS and both the period of hospitalization at the pediatric intensive care unit and the total length of hospital stay. CONCLUSIONS: Pedestrian accidents caused by motor vehicles in children playing at the street contributed most significantly to MT in the urban pediatric population. Special care for prevention must be taken in the age group of 6 to 10 years. Head injury was the main cause of death in children with multiple trauma. MISS was found to be a good predictor of survival and duration of hospital stay in pediatric MT. PMID- 11051156 TI - Effect of PEEP and suction via chest drain on functional residual capacity and lung compliance after surgical repair of congenital diaphragmatic hernia: preliminary observations in 5 patients. AB - BACKGROUND/PURPOSE: Congenital diaphragmatic hernia (CDH) is associated with pulmonary hypoplasia that limits survival. The authors' knowledge on lung mechanics and lung volumes in these patients with hypoplastic lungs is still limited. Therefore, the authors performed measurements of functional residual capacity (FRC), compliance of the respiratory system (CRS), and tidal volume in 5 full-term infants (gestational age, 38 to 40 weeks; birth weight, 2,800 to 3,530 g) before and after surgical repair of neonatal CDH. METHODS: The authors studied the influence of different levels of positive end-expiratory pressure (PEEP) and suction via inserted ipsilateral chest tube connected to a water seal on lung volume and lung mechanics. A computerized tracer gas (SF6) washout method was used for serial measurements of FRC. Compliance of the respiratory system was determined according to insufflatory method. RESULTS: The authors found a preoperative compliance between 1.5 and 3.9 mL/kPa/kg and a preoperative FRC between 9.1 and 12.9 mL/kg indicating severe hypoplasia of the lungs in all patients. Immediately after surgical repair of CDH, compliance decreased to 85% (78% to 91%) of preoperative value, and FRC increased to 132% (110% to 150%) of preoperative value under mechanical ventilation while at 4 cm of water of PEEP and at -10 cm of water of suction via chest drain with the need of high fraction of inspired oxygen. After reduction of PEEP from 4 to 2 or 1 cm of water and lowering suction from -10 cm of water to -2 or 0 cm of water FRC decreased to 103% (80% to 122%) of preoperative value and compliance, and tidal volume improved to 135% (110% to 147%) of preoperative value resulting in increased alveolar ventilation, correction of acidosis and improvement in oxygenation. During the first days after surgery inadequate high PEEP or strong suction via chest tube drainage resulted in increase in FRC paralleled by decrease in compliance indicating overdistension of these hypoplastic lungs. CONCLUSIONS: The data show that overdistension of hypoplastic lungs in infants with CDH can be detected and excluded by repeated measurements of FRC and compliance in these critical ill infants. These data might help setting appropriate ventilator parameters, adequate suction via chest drain, and thereby improve gas exchange and outcome. PMID- 11051157 TI - Successful fetal sacrococcygeal teratoma resection in a hydropic fetus. AB - BACKGROUND/PURPOSE: The development of hydrops in a fetus with a sacroccocygeal teratoma (SCT) usually is a predictor of fetal demise; in utero resection may offer the only chance of survival. Although the authors had performed this procedure in 3 previous cases, they had no long-term patient survival. The authors report a successful case of in utero resection of a fetal sacrococcygeal teratoma. METHODS: The authors resected a fetal SCT from a 23-weeks-gestation hydropic fetus, using gradually tightening umbilical tapes at the tumor base, electrocautery, and careful sharp dissection. After a blood transfusion, the fetus suffered cardiac arrest but was resuscitated and returned to the uterus. RESULTS: Postoperatively, residual SCT growth ceased, and hydrops rapidly resolved. Five weeks after the procedure, the infant was delivered because of preterm labor, and, after resection of residual SCT, was discharged home at 3 months of age. She is now a healthy 3 year old. CONCLUSION: This case shows that successful fetal SCT resection and long-term patient survival is possible. PMID- 11051158 TI - Internal inguinal hernia in a 20-month-old boy. AB - The authors report on a 20-month-old boy with internal inguinal hernia. The patient was reoperated for clinical signs of intestinal obstruction after an incarcerated inguinal hernia repair. The diagnosis of internal inguinal hernia was proven by surgical exploration. This is the first internal inguinal hernia case reported in the literature and completely different from the atypical parainguinal hernias reported before. PMID- 11051159 TI - Salmonella typhimurium: a rare cause of colonic ulceration and perforation in infancy. AB - A rare case of a healthy infant with colonic ulcers caused by Salmonella typhimurium infection that presented with colonic perforation, hypovolemic, and septicemic shock is discussed. It stresses the importance of considering an infective process such as salmonellosis in the differential diagnosis of colonic ulceration in an infant and illustrates the unique histologic finding of colonic inflammatory changes with sparing of the small intestine. PMID- 11051160 TI - Invasive Candida enteritis of the newborn. AB - Three premature infants (<800 g) showed invasive Candida at the site of their intestinal perforations. This entity is distinct from Candida peritonitis complicating necrotizing enterocolitis and was uniformly fatal. Recognition and aggressive antifungal therapy may improve outcomes. PMID- 11051161 TI - Aplasia of the right lung in a 4-year-old child: surgical stabilization of the mediastinum by diaphragm translocation leading to complete recovery from respiratory distress syndrome. AB - Lung aplasia is defined as unilateral absence of the lung with preservation of main bronchus remnant at the tracheal bifurcation. Patients usually die soon after birth and there is no specific therapy for this condition, as evidenced by the literature. The authors present a case of an infant that was asymptomatic with this malformation until 3 months of age, when the child had respiratory distress syndrome. Subsequently, lung aplasia was diagnosed. The authors performed an extrapleural dissection and cephalad translocation of diaphragm to reduce the mediastinal shift and heart rotation, to relieve a kink and compression of the trachea by the aortal arch and truncus arteriosus, as well as to relieve hyperinflation of lung parenchyma and provide recovery from respiratory distress syndrome. This new approach resulted in complete recovery from respiratory distress syndrome and full tolerance of physical exercise. The child underwent follow-up for 4 years. Diaphragmatic translocation may be useful in treatment of respiratory disorders associated with lung aplasia. PMID- 11051162 TI - Conservative management of hepatic duct injury after blunt trauma: a case report. AB - Nonoperative management of solid organ injury in children with blunt abdominal trauma represents the standard of care. In rare cases, a major duct injury with persistent bile leakage may result from blunt trauma to the liver. This injury is of concern in patients treated nonoperatively because it generally must be treated with major abdominal surgery. The authors describe a case of hepatic duct injury from blunt trauma in which healing occurred without surgical repair or resection. PMID- 11051163 TI - Salivary gland choristoma of the anterior chest wall. AB - Salivary gland choristoma (heterotopic salivary gland tissue) is a rare condition that occurs at various locations within the head and neck. Diagnostic criteria and embryogenesis of this entity remain unclear. Presented herein is the first reported case of salivary gland choristoma on the anterior chest wall. Surgical treatment is recommended. PMID- 11051164 TI - Intrauterine torsion of a wandering spleen presenting as an abdominal cystic swelling. AB - Wandering spleen is a rare clinical condition that presents commonly with splenic infarction secondary to torsion. Intrauterine torsion of a wandering spleen, however, is extremely rare. An unusual case of intrauterine torsion of a wandering spleen presenting as an abdominal mass is reported. PMID- 11051165 TI - Is surgical treatment of lipoblastoma always necessary? AB - Lipoblastoma is an uncommon, benign mesenchymal tumor with an excellent prognosis despite its potential to local invasion and rapid growth. However, in the literature, a spontaneous resolution has never been reported, and, consequently, the need for a complete surgical excision has never been questioned. The authors report a case of a 2-day-old boy with congenital diffuse lipoblastoma in the left thigh, which forced us to withhold from surgical treatment to avoid the risk of mutilation in a patient so young. The lesion was followed-up by imaging, and a complete spontaneous resolution of the diffuse lipoblastoma was shown by magnetic resonance imaging (MRI) at 1-year follow-up. In the literature, a complete surgical excision is recommended. The results of this case suggest that a "wait and see" approach is justified at least in infants with huge invasive lesions requiring a mutilating excision. PMID- 11051166 TI - Inverted papilloma of the urinary bladder in children: case report and review of prognostic significance and biological potential behavior. AB - Inverted papilloma of the urinary bladder is rare in the pediatric population. Despite several reports in the literature the prognostic significance and biological potential behavior of this lesion remain uncertain. The authors report a case of polypoid inverted papilloma of the urinary bladder in an 11-year-old boy and review its pathology. The pediatric population with this lesion is an ideal group to provide intense, long-term follow-up to define the biological behavior and prognosis significance of this lesion. PMID- 11051167 TI - Intrathyroid parathyroid gland and neonatal primary hyperparathyroidism. AB - Neonatal hyperparathyroidism (NPHP) is exceedingly rare and often fatal. A neonate is presented with a serum calcium concentration of 33 mg/dL, an intrathyroid parathyroid gland, and a family history of hypocalciuric hypercalcemia (FHH). She underwent successful total parathyroidectomy. Six years later, the child is normocalcemic and developmentally normal, requiring calcium and calcitrol replacement. The results of this case support the concept that NPHP is associated with parathyroid hyperplasia and is part of a continuum that includes FHH. PMID- 11051168 TI - Chronic recurrent multifocal osteomyelitis associated with ulcerative colitis: a case report. AB - Chronic recurrent multifocal osteomyelitis (CRMO) is a rare disease of bone first described by Giedion et al in 1972. It is associated with several pathologic processes including psoriasis, palmoplantar pustulosis, and SAPHO syndrome (synovitis, acne, pustulosis, hyperostosis, and osteitis). The only published association of CRMO and Crohn's disease was reported by Bognar et al in 1998. The authors describe the association of CRMO and ulcerative colitis (UC) in a 12-year old girl. As far as the authors know, this is the first published report of CRMO associated with UC and the second of CRMO associated with inflammatory bowel diseases. PMID- 11051169 TI - An unusual case of epigastric heteropagus: parasite with a rudimentary heart. AB - Parasitic or heterotopic conjoined twins are exceedingly rare, and these cases are referred to as "heteropagus" when there is a parasitic attachment in a nonduplicated fashion to any portion of the body. Epigastric heteropagus twinning refers to the attachment of the parasite to the epigastric region of the autosite. An unusual epigastric heteropagus case is presented with a rudimentary cardiopulmonary and also nearly complete gastrointestinal and genitourinary system of the parasite, and an organogenetic and vascular status of the previously reported cases are reviewed. The current case might be unique for the parasite having a cardiopulmonary development--although rudimentary--and this might be the reason that it has more complete organogenesis than the cases that have been presented previously in the literature. PMID- 11051170 TI - Intraoperative pancreatoscopy for pancreatic duct stone debris distal to the common channel in choledochal cyst. PMID- 11051171 TI - Delayed splenic rupture following low impact activity. PMID- 11051172 TI - Recurrent pancreatitis and pancreas divisum cured by sphincteroplasty of both major and minor papillae. PMID- 11051184 TI - Robotics, technology, and the future of surgery. PMID- 11051185 TI - Laparoscopic adhesiolysis and relief of chronic pelvic pain. AB - OBJECTIVE: To evaluate the short- and long-term results of laparoscopic enterolysis in patients with chronic pelvic pain following hysterectomy. METHODS: Forty-eight patients were evaluated at time intervals from 2 weeks to 5 years after laparoscopic enterolysis. Patients were asked to rate postoperative relief of their pelvic pain as complete/near complete relief (80-100% pain relief), significant relief (50-80% pain relief), or less than 50% or no pain relief. RESULTS: We found that after 2 to 8 weeks, 39% of patients reported complete/near complete pain relief, 33% reported significant pain relief, and 28% reported less than 50% or no pain relief. Six months to one year postlaparoscopy, 49% of patients reported complete/near complete pain relief, 15% reported significant pain relief, and 36% reported less than 50% or no pain relief. Two to five years after laparoscopic enterolysis, 37% of patients reported complete/near complete pain relief, 30% reported significant pain relief, and 33% reported less than 50% or no pain relief. Some patients required between 1 and 3 subsequent laparoscopic adhesiolysis. A total of 3 enterotomies and 2 cystotomies occurred, all of which were repaired laparoscopically. CONCLUSION: We conclude that laparoscopic enterolysis may offer significant long-term relief of chronic pelvic pain in some patients. PMID- 11051186 TI - Surgical strategy of complex empyema thoracis. AB - BACKGROUND: The optimal treatment of empyema thoracis has been widely debated. Proponents of pleural drainage alone, drainage plus fibrinolytic therapy, video assisted thoracoscopic surgical (VATS) debridement, and open thoracotomy each champion the efficacy of their approach. METHODS: This study examines treatment of complex empyema thoracis between June 1, 1994, and April 30, 1997. Twenty-one men and 9 women underwent 30 drainage/decortication procedures (14 open thoracotomies and 16 VATS) in treatment of their disease. Effusion etiology was distributed as follows: infectious-14; neoplastic-associated-7; traumatic-3; other-6. RESULTS: The mean preoperative hospital stay was 14 +/- 8.8 days, (11.4 +/- 6.5 days for VATS vs 16.8 +/- 10.2 days for thoracotomy). Hospital stay from operation to discharge for thoracotomy patients was 10.0 -/+ 7.2 days (median 8.5 days) and for VATS patients 17.6 -/+ 16.8 days (median 11 days). These differences were not statistically significant. Duration of postoperative thoracostomy tube drainage was 8.3 -/+ 4.6 days for thoracotomy patients and 4.7 /+ 2.8 days in the VATS group (p = 0.01). Operative time for the thoracotomy group was 125.0 -/+ 71.7 minutes, while the VATS group time was only 76.2 -/+ 30.7 minutes. Estimated blood loss for the thoracotomy group was 313.9 -/+ 254.0 milliliters and for the VATS group 131.6 -/+ 77.3 milliliters. Three of the 30 patients (10.0%) required prolonged ventilator support (>24 hours). Morbidity included one diaphragmatic laceration (VATS group) and one thoracic duct laceration (thoracotomy). Two VATS procedures (6.7%) required conversion to open thoracotomy for thorough decortication. CONCLUSIONS: The surgical approach to empyema thoracis is evolving. In the absence of comorbid factors, the significantly lower requirement for chest tube drainage time in the VATS patients suggests that this modality is an attractive alternative to thoracotomy in the treatment of complex empyema thoracis. PMID- 11051187 TI - Influence of pneumoperitoneum on the deep venous system during laparoscopy. AB - BACKGROUND: There is widespread concern that laparoscopic procedures that are usually performed under general anesthesia, using muscle relaxation, in a reverse Trendelenberg position and with pneumoperitoneum, may lead to venous stasis in lower limbs. OBJECTIVE: To evaluate perioperative changes in the venous system and determine the frequency of deep venous thrombosis associated with minimally invasive surgery. DESIGN: Prospective consecutive series. SUBJECTS: Sixty-five patients undergoing elective minimally invasive surgery. INTERVENTION: Laparoscopic procedures with no thromboprophylaxis. RESULTS: Sixty-one patients completed the investigations (coagulation profile and lower limb venous duplex scan) on admission and on the first postoperative day. The median duration of pneumoperitoneum was 45 minutes (range: 18-90 minutes). None of postoperative scans revealed thrombosis. No significant changes in the postoperative coagulation profile were identified. Perioperative scans of the left femoral vein revealed an increase in cross-sectional area (P<0.05) and a decrease in peak blood velocity (P<0.05). CONCLUSION: In this study of low-risk patients for thromboembolism, laparoscopy with pneumoperitoneum at pressures below 12 mm Hg per se did not increase the prevalence of deep venous thrombosis. This implies that venous hemodynamic changes observed during pneumoperitoneum did not cause deleterious venous stasis. Still, caution needs to exercised with regard to the view that no special precautions to prevent deep venous thrombosis are warranted in patients undergoing laparoscopy. PMID- 11051188 TI - Sexual activity as cause for non-surgical pneumoperitoneum. AB - BACKGROUND: Pneumoperitoneum is usually seen after bowel perforations and surgical procedures. An increasing number of cases of non-surgical pneumoperitoneum related to sexual activity has been reported worldwide over the last years. CASE EXAMPLE: A typically young, otherwise healthy woman comes into the emergency department of Stanford University, California, complaining of recurrent chest pain. Free air under the diaphragm disclosed in the X-ray usually leads to intensive, costly and invasive diagnostics sometimes resulting in emergency laparotomy without any results. Finally, after thorough discussion of the sexual history of the patient is taken, vaginal insufflation during sexual activity is revealed as the cause of non-surgical pneumoperitoneum. DISCUSSION: Patients are often unaware of the open access between the vagina and abdomen. Insufflation pressure during vaginal insufflation with >100 mm Hg--used as a diagnostic tool in CO2-pertubation--can dilate genital organs and push remarkable amounts of air into the abdomen. Gas resorption can take up to several days, and the patient often does not connect the pain to its cause. Embarrassment and modesty often prevent the patient from talking about sexual activity. CONCLUSION: Sexual pneumoperitoneum is not a bizarre sex accident but a rare and serious patho-mechanism. In cases of atypical non-surgical pneumoperitoneum in sexually active women, a careful inquiry into the medical-sexual history can reveal the cause of pathophysiology without comprehensive, painful and unnecessary diagnostics. Sexual history as a diagnostic tool should always be considered in unclear cases. PMID- 11051189 TI - Intraperitoneal bupivacaine does not attenuate pain following laparoscopic cholecystectomy. AB - BACKGROUND: Laparoscopic cholecystectomy is characterized by a short hospital stay. Hence, pain control on the day of surgery is increasingly important. The aim of this study was to evaluate the effect of intraperitoneal bupivacaine on pain relief following laparoscopic cholecystectomy. METHODS: Sixty patients undergoing elective laparoscopic cholecystectomy were prospectively randomized into 2 groups. Following removal of the gallbladder, group A received 100 mg of bupivacaine in 50 cc of saline, installed into the gallbladder bed and right subphrenic space. Group B received saline without bupivacaine. Pain was assessed using a visual/analog scale at fixed-time intervals. RESULTS: No significant difference occurred in the average pain levels between the groups at 1, 2, 4, and 14 hours postsurgery. The average analgesic requirement was lower in the bupivacaine group, but this did not reach statistical significance. CONCLUSION: Application of intraperitoneal bupivacaine did not attenuate pain following laparoscopic cholecystectomy, and no role exists for its routine use. PMID- 11051190 TI - Laparoscopic splenectomy in a patient with acquired angioneurotic edema. AB - BACKGROUND: We report the case of a 77-year-old female with acquired angioneurotic edema, C1 esterase inhibitor level = 4mg/dL, who was scheduled to undergo laparoscopic splenectomy. METHODS: In the operating room, we administered on call 500 units (UI) of C1 esterase inhibitor concentrate intravenously. Intraoperative hemodynamic instability and generalized blood oozing improved following the administration of aprotinin 250000 UI intravenous (IV) drip. CONCLUSION: We recommend the administration of an antifibrinolytic agent in addition to C1 esterase inhibitor concentrate in patients with acquired angioneurotic edema. PMID- 11051191 TI - Laparoscopic cystogastrostomy for pancreatic pseudocyst: a case report. AB - A 49-year-old man with a history of acute pancreatitis was hospitalized with a diagnosis of pancreatic pseudocyst. Ultrasonography, computed tomography, and magnetic resonance imaging all demonstrated a homogeneous cyst, 9 x 4 cm in size, at the tail of the pancreas without mural nodules or septa. Because an intestinal structure was identified between the cyst and stomach preoperatively by computed tomography and endoscopic ultrasonography, laparoscopic cystogastrostomy was carried out instead of percutaneous or endoscopic cyst drainage. The cyst was exposed by dissecting the lesser omentum and found to have no adhesion to the surrounding tissues. Anastomosis was performed using an endoscopic linear stapler via small cystotomy and gastrotomy openings on the lesser curvature, which were then sutured laparoscopically. The postoperative course was uneventful. Laparoscopic surgery is recommended as a safe, reliable, and minimally invasive treatment for managing pancreatic pseudocyst. PMID- 11051192 TI - Aging and longevity genes. AB - The genetics of aging has made substantial strides in the past decade. This progress has been confined primarily to model organisms, such as filamentous fungi, yeast, nematodes, fruit flies, and mice, in which some thirty-five genes that determine life span have been cloned. These genes encode a wide array of cellular functions, indicating that there must be multiple mechanisms of aging. Nevertheless, some generalizations are already beginning to emerge. It is now clear that there are at least four broad physiological processes that play a role in aging: metabolic control, resistance to stress, gene dysregulation, and genetic stability. The first two of these at least are common themes that connect aging in yeast, nematodes, and fruit flies, and this convergence extends to caloric restriction, which postpones senescence and increases life span in rodents. Many of the human homologs of the longevity genes found in model organisms have been identified. This will lead to their use as candidate human longevity genes in population genetic studies. The urgency for such studies is great: The population is graying, and this research holds the promise of improvement in the quality of the later years of life. PMID- 11051193 TI - Antioxidant defense in centenarians (a preliminary study). AB - The study was designed to assess the antioxidant defense mechanisms, either enzymatic or non-enzymatic, in a group of sixteen centenarians (one male and fifteen female subjects aged 101 to 105 years) living in the Upper Silesia district (Poland) in order to evaluate the potential role of antioxidant defenses in human longevity. The results of our preliminary study showed that in comparison with young healthy female adults the centenarians had significantly higher red blood cell glutathione reductase and catalase activities and higher, although insignificantly, serum vitamin E level. PMID- 11051194 TI - Proliferation and apoptosis of human T cells during replicative senescence--a critical approach. AB - Normal human T lymphocytes growing in culture undergo replicative senescence. Previously, we have shown that in our conditions polyclonal T cells cease proliferation after about three weeks (Radziszewska et al., 1999, Cell Biol. Int. 23, 97-103). Now we present results of a more detailed analysis of in vitro growth as well as phenotypic changes of T cells. Cell cycle analysis showed that about 20% of cells were in the S phase until the 17th day of culture (young cells). The highest number of mitotic cells (phase G2/M; 10%) was observed during the first week of culture. All not dividing senescent cells were stopped in the G1 phase (after the 30th day of culture). The sub-G1 fraction which represents apoptotic cells did not exceed 8% during the whole period until the 30th day of culture. During in vitro T-cell growth, a rather rapid selection to CD3+ CD8+ cells occurs. In the presenescent (between the 17th and 30th day) and senescent populations the majority of cells (above 90%) were CD8 positive. We also have checked the expression of alpha-chain interleukin-2 (IL-2) receptor (CD25). In young and presenescent cells about one third of cells was CD25 positive, but only 15% in the pool of senescent cells. Immunoblotting analysis of p16 protein recognized previously as a marker of senescent T cells, showed its highest and transient expression in presenescent cells. A critical review of the polyclonal T cell replicative senescence model is presented. PMID- 11051195 TI - Compensatory effect of TNFalpha on low natural killer activity in the elderly. AB - Regulatory effect of CD25, an activation antigen the alpha subunit of interleukin 2 receptor (IL2R) on the activity of natural killer (NK) cells was studied in fifty elderly (57-70 years old) and fifty young people (19-35 years old). Cytotoxic NK activity was assessed by 51Cr release assay, the levels of interleukin 2 (IL2) and tumour necrosis factors alpha (TNFalpha) were measured using bioassays and expression of CD16 and CD25 proteins by flow cytometry. Low NK activity in the elderly was associated with decline of full health, lowered serum concentration of IL2 and increased production of TNFalpha during NK reaction. Inhibition of TNFalpha activity by anti-TNF monoclonal antibody suppressed exclusively NK activity of low NK responders. Moreover, stimulation in vitro of blood mononuclear cells, with TNFalpha induced in the elderly low NK responders a significantly higher increase of the CD25 expression on the surface of NK cells as compared with that in the elderly high responders. Since the CD25 molecule constitutes a subunit of the high affinity receptor, binding IL2 to immunocompetent cells, its increased expression on NK cells of low NK responders would enable them to bind even low amounts of the endogenous IL2 available in this group of the elderly. Thus, an overproduction of TNFalpha seems to be a mechanism compensating, in the non-fully healthy elderly, for the decreased IL2 production, promoting efficient cytotoxic reaction. PMID- 11051196 TI - Does senile impairment of cholinergic system in rats concern only disturbances in cholinergic phenotype or the progressive degeneration of neuronal cell bodies? AB - The trophic effect of continuous intraventricular infusion of nerve growth factor (NGF) on morphology of the basal forebrain (BF) cholinergic neurons was tested in 4- and 28-month-old male Wistar rats. All studies were conducted using behaviorally uncharacterized animals from the same breeding colony. Immunohistochemical procedure for choline acetyltransferase (ChAT) and p75NTR receptor has been applied to identify cholinergic cells in the structures of basal forebrain (BF). Using a quantitative image analyzer, morphometric and densitometric parameters of ChAT- and p75NTR-positive cells were measured immediately after cessation of NGF infusion. In 28-month-old non-treated rats the number of intensively ChAT-positive cells in all forebrain structures was reduced by 50-70% as compared with young animals. The remaining ChAT-positive cells appeared shrunken and the neuropil staining was NTR markedly reduced. In contrast, the same neurons when stained for p75 were numerous and distinctly visible with perfect morphology. Analysis of Nissl stained sections also showed that 28-month-old rats did not display significant losses of neuronal cell bodies. NGF restored the number of intensely stained ChAT-positive cells to about 90% of that for young controls and caused a significant increase in size of those cells in 28-month-old rats as compared with the control, age-matched group. NGF did not influence the morphology of p75NTR-positive neurons, which were well labeled, irrespective of treatment and age of the rats. In 4-month-old rats, NGF infusion decreased the intensity of both ChAT and p75NTR immunostaining. These data provide some evidence for preservation of BF cholinergic neurons from atrophy during aging and indicate that senile impairment of the cholinergic system in rats concerns decrease in ChAT-protein expression rather than an acute degeneration of neuronal cell bodies. Treatment with NGF resulted in restoration of cholinergic phenotype in the BF neurons of aged rats. However, the present study also rises issue of possible detrimental effects of NGF in young normal animals. PMID- 11051197 TI - Age-related alteration of poly(ADP-ribose) polymerase activity in different parts of the brain. AB - Poly(ADP-ribose) polymerase (PARP) is a conserved enzyme involved in the regulation of DNA repair and genome stability. The role of PARP during aging is not well known. In this study PARP activity was investigated in nuclear fractions from hippocampus, cerebellum, and cerebral cortex of adult (4 months), old adult (14 months) and aged (24-27 months) rats. Concomitantly, the free radical evoked lipid peroxidation was estimated as thiobarbituric acid reactive substances (TBARS). The specific activity of PARP in adult brain was about 25, 21 and 16 pmol/mg protein per min in hippocampus, cerebellum and cerebral cortex, respectively. The enzyme activity was higher in all investigated parts of the brain of old adults. In aged animals PARP activity was lower in hippocampus by about 50%, and was unchanged in cerebral cortex and in cerebellum comparing to adult rats. The concentration of TBARS was the same in all parts of the brain and remained unchanged during aging. There is no direct correlation between PARP activity and free radical evoked lipid peroxidation during brain aging. The lowered enzyme activity in aged hippocampus may decrease DNA repair capacity which subsequently may be responsible for the higher vulnerability of hippocampal neurons to different toxic insults. PMID- 11051198 TI - Effect of aging on UVC-induced apoptosis of rat splenocytes. AB - UVC-induced apoptotic symptoms such as morphological changes, DNA fragmentation, Bcl-2 and Bax protein expression were examined in primary splenocyte cultures from young (3 months) and old (24 months) rats. The activities of AP-1 and CRE transcription factors in UVC-irradiated splenocytes were also assessed. At 24 h after UVC irradiation 40% of cells derived from young rats were found to be apoptotic, which was twice as much as in splenocytes from old rats. Apoptosis in cells from old rats did not give typical symptoms like a "DNA ladder" and Bcl-2 protein downregulation, in contrast to splenocytes from young rats. No AP-1 transcription factor activity was found in UVC-irradiated splenocytes from old animals and only a trace activity in splenocytes from young animals. This indicates that, UVC-induced apoptosis in rat splenocytes is practically AP-1 independent and that cells from old rats are less sensitive to UVC irradiation than splenocytes from young rats. PMID- 11051199 TI - The effects of galactosamine on UTP levels in the livers of young, adult and old rats. AB - Galactosamine (GalN), a well-known hepatotoxin that depletes the cellular pool of uracil nucleotides, was previously shown to have greater impact on the inhibition of protein synthesis in hepatocytes of old rats as compared with young animals (Kmiec 1994, Ann. N.Y. Ac. Sci. 717, 216-225). In the present study we compared the effects of GalN on the nucleotide content (measured by ion-exchange HPLC) in the livers of young (4 months), adult (12 months), and old (24-26 months old) rats two hours after its intraperitoneal administration. UTP content of the livers of old control rats was significantly lower (by 28%) than that of young animals. GalN administration decreased the UTP content in the livers of young, adult and old rats by, respectively, 55%, 65% and 89%, and increased the content of UDP-sugars by 189%, 175% and 305%. The hepatic content of ATP, ADP, AMP, NAD, GTP except CTP did not differ significantly among the age groups of rats studied, and was not changed by GalN treatment. The content of CTP was significantly higher in old rats (P < 0.03) upon GalN treatment. The lower hepatic content of UTP may partially explain the increased sensitivity of hepatocytes and livers of old rats to the action of galactosamine, and possibly to other hepatotoxic compounds that decrease transcription in the liver. PMID- 11051200 TI - Effect of stress on the life span of the yeast Saccharomyces cerevisiae. AB - A correlation is known to exist in yeast and other organisms between the cellular resistance to stress and the life span. The aim of this study was to examine whether stress treatment does affect the generative life span of yeast cells. Both heat shock (38 degrees C, 30 min) and osmotic stress (0.3 M NaCl, 1 h) applied cyclically were found to increase the mean and maximum life span of Saccharomyces cerevisiae. Both effects were more pronounced in superoxide dismutase-deficient yeast strains (up to 50% prolongation of mean life span and up to 30% prolongation of maximum life span) than in their wild-type counterparts. These data point to the importance of the antioxidant barrier in the stress-induced prolongation of yeast life span. PMID- 11051201 TI - In vitro expression analysis of R68G and R68S mutations in phenylalanine hydroxylase gene. AB - Phenylketonuria (PKU), an autosomal recessive disorder caused be a deficiency of hepatic phenylalanine hydroxylase (PAH), is clinically very heterogeneous. At the molecular level, more than 400 mutations in the PAH gene are known to date, which in different genotype combinations could account for biochemical and clinical variability of symptoms. In vitro expression studies on R68G and R68S mutations causing mild phenylketonuria are presented. PMID- 11051202 TI - Characterization and expression analysis of the yellow lupin (Lupinus luteus L.) gene coding for nodule specific proline-rich protein. AB - The LlPRP2 gene coding for a proline-rich protein shows a high level of similarity to, as well as significant differences from the family of ENOD2 nodule specific genes. Several sequence motifs with putative regulatory function were identified in the 5' and 3' noncoding regions of the LlPRP2 gene. Northern blot analysis revealed that the expression of the LlPRP2 gene begins 9 days after inoculation of yellow lupin roots with Bradyrhizobium sp. (Lupinus); the expression is restricted to symbiotic nodules and is not detected in other tissues or organs. Detailed hybridization analysis showed that, when expression is activated, the LlPRP2 transcript is modified so as to produce at least three bands and a continuous distribution of decay intermediates. The modification of the LlPRP2 transcript probably involves degradation from the 5'- and/or 3'-ends of the RNA molecules. Southern blot analysis indicates that only one gene is present in the yellow lupin genome. The presence of genes homologous to the LlPRP2 gene was confirmed for three cultivars of yellow lupin and for Lupinus angustifolius. However, LlPRP2 homologues were not detected in Lupinus albus cv. Bac, indicating that this plant may lack the ENOD2 sequence. PMID- 11051203 TI - Direct transfer of IL-12 gene into growing Renca tumors. AB - We investigated the feasibility of transferring naked plasmid DNA containing a therapeutic gene (IL-12) into mice harboring growing Renca tumors. We found that naked DNA transferred into growing Renca and B16(F10) tumors gives higher expression level of reporter gene than complexes of DNA with DDAB/DOPE or DC Chol/DOPE. Transfer of naked DNA carrying the IL-12 gene into growing Renca tumors causes a distinct therapeutic effect that depends on the time span between inoculation of mice with cancer cells and the beginning of the therapy. Therapy started on day 3 resulted in total cure (100%) of mice. PMID- 11051205 TI - Comparative biochemical analysis of lectin and nuclease from Chelidonium majus L. AB - It has been recently recognized that lectins exhibit other activities besides hemagglutination. Previously we have found that purified lectin from Chelidonium majus showed DNase activity (Fik, Gozdzicka-Jozefiak & Kedzia, 1995, Herba Polon. 41, 84-95). Comparison of lectin and DNase from the sap from leaves and roots of Chelidonium majus proved that both these compounds are composed of 24 kDa monomer subunits which have an identical N-terminal sequence but differ in amino-acid composition and degree of glycosylation. Possible interrelationship between lectin and DNase is discussed. PMID- 11051204 TI - Selectin glycoprotein ligands. AB - Lectin selectins and their counter-receptors participate in discontinuous cell cell interactions concurrent with leukocyte tethering and rolling on endothelium, which, in consequence, leads to leukocyte penetration to lymphatic organs and generation of inflammation sites. Counter-receptors are glycoproteins in which carbohydrate units, the direct selectin ligands, are built into the polypeptide framework. In this review, the distribution, structure and function of the main ligands and counter-receptors for P-, L- and E-selectins known so far, have been discussed. The common biosynthetic pathway of sialyl-Lewis x and sulpho-sialyl Lewis x determinants of selectin ligands has been described. PMID- 11051206 TI - The participation of ribosome-UDP-GalNAc complex in the initiation of protein glycosylation in vitro. AB - The gastric epithelial cells ribosome-UDP-GalNAc complex is a donor of UDP-GalNAc as the substrate for N-acetylgalactosaminyltransferase, which catalyse the transfer of GalNAc residue to the polypeptide, existing on polysomes. It was observed that the deglycosylated porcine mucin and synthetic peptide (PTSSPIST) can be also glycosylated with participation of N-acetylgalactosaminyltransferase and ribosome-UDP-GalNAc complex. The probability of the ribosome-UDP-GalNAc complex as an intermediate in the O-glycosylation is considered. PMID- 11051207 TI - Differences of alpha3beta1 integrin glycans from different human bladder cell lines. AB - Expression as well as properties of integrins are altered upon transformation. Cell adhesion regulated by integrins is modulated by glycosylation, one of the most frequent biochemical alteration associated with tumorogenesis. Characterisation of carbohydrate moieties of alpha3beta1 integrin on the cultured human bladder carcinoma (T-24, Hu456, HCV 29T) and human normal ureter and bladder epithelium (HCV 29, Hu609) cell lines was carried out after an electrophoresis and blotting, followed by immunochemical identification of alpha3 and beta1 integrin chains and analysis of their carbohydrates moieties using highly specific digoxigenin-labelled lectins. In all the studied cell lines alpha3beta1 integrin was glycosylated although in general each subunit differently. Basic structures recognized in beta1 subunit were tri- or tetraantennary complex type glycans in some cases sialylated (T-24, HCV 29, HCV 29T) and fucosylated (Hu609, HCV 29T). Positive reaction with Phaseolus vulgaris agglutinin and Datura stramonium agglutinin suggesting the presence of beta1-6 branched N-linked oligosaccharides was found in cancerous cell lines (T-24, Hu456) as well as in normal bladder epithelium cells (Hu609). High mannose type glycan was found only in beta1 subunit from Hu456 transitional cell cancer line. On the other hand alpha3 subunit was much less glycosylated except the invasive cancer cell line T-24 where high mannose as well as sialylated tri- or tetraantennary complex type glycans were detected. This observation suggests that changes in glycosylation profile attributed to invasive phenotype are rather associated with alpha3 not beta1 subunit. PMID- 11051208 TI - Human diadenosine triphosphate hydrolase: preliminary characterisation and comparison with the Fhit protein, a human tumour suppressor. AB - Human platelets diadenosine triphosphatase was characterised and compared with the Fhit protein, a human tumour suppressor with diadenosine triphosphatase activity. Both enzymes exhibit similar Km, are similarly activated by Mg2+, Ca2+ and Mn2+, and inhibited by Zn2+ and suramin. However, they are differentially inhibited by Fhit antibodies and exhibit differences in gel-filtration behaviour. PMID- 11051209 TI - Detection of damage-recognition proteins from human lymphocytes. AB - Proteins recognizing and binding to damaged DNA (DDB-proteins) were analyzed in human lymphocytes obtained from healthy donors. Using an electrophoretic mobility shift assay several complexes between nuclear extract proteins and damaged DNA were detected: a complex specific for DNA damaged by N-acetoxy-N acetylaminofluorene, another complex specific for UV-irradiated DNA, and two complexes specific for DNA damaged by cis-dichlorodiammine platinum. All the detected complexes differed in electrophoretic mobility and possibly contained different proteins. Complexes specific for free DNA ends were also detected in protein extracts from lymphocytes. PMID- 11051210 TI - A new look at adaptive mutations in bacteria. AB - This is a short survey of the adaptive mutation processes that arise in non- or slowly-dividing bacterial cells and includes: (i) bacterial models in which adaptive mutations are studied; (ii) the mutagenic lesions from which these mutations derive; (iii) the influence of DNA repair processes on the spectrum of adaptive mutations. It is proposed that in starved cells, likely as during the MFD phenomenon, lesions in tRNA suppressor genes are preferentially repaired and no suppressor tRNAs are formed as a result of adaptive mutations. Perhaps the most provocative proposal is (iv) a hypothesis that the majority of adaptive mutations are selected in a pre-apoptotic state where the cells are either mutated, selected, and survive, or they die. PMID- 11051211 TI - Reversion of argE3 ochre strain Escherichia coli AB1157 as a tool for studying the stationary-phase (adaptive) mutations. AB - Adaptive (starvation-associated) mutations occur in non-dividing cells and allow growth under the selective conditions imposed. We developed a new method for the determination of adaptive mutations in Escherichia coli. The system involves reversion to prototrophy of the argE3OC mutation and was tested on AB1157 strains mutated in the mutT and/or mutY genes. The bacteria that mutated adaptively grow into colonies on minimal medium plates devoid of arginine (starvation conditions) when incubated longer than 4 days. Using the replica plating method we solved the problem of discrimination between growth-dependent and adaptive argE3-->Arg+ revertants. Phenotype analysis and susceptibility of the Arg+ revertants to a set of T4 phage mutants create an additional possibility to draw a distinction between these two types of Arg+ revertants. PMID- 11051212 TI - Ubiquinone. Biosynthesis of quinone ring and its isoprenoid side chain. Intracellular localization. AB - Ubiquinone, known as coenzyme Q, was shown to be the part of the metabolic pathways by Crane et al. in 1957. Its function as a component of the mitochondrial respiratory chain is well established. However, ubiquinone has recently attracted increasing attention with regard to its function, in the reduced form, as an antioxidant. In ubiquinone synthesis the para-hydroxybenzoate ring (which is the derivative of tyrosine or phenylalanine) is condensed with a hydrophobic polyisoprenoid side chain, whose length varies from 6 to 10 isoprene units depending on the organism. para-Hydroxybenzoate (PHB) polyprenyltransferase that catalyzes the condensation of PHB with polyprenyl diphosphate has a broad substrate specificity. Most of the genes encoding (all-E)-prenyltransferases which synthesize polyisoprenoid chains, have been cloned. Their structure is either homo- or heterodimeric. Genes that encode prenyltransferases catalysing the transfer of the isoprenoid chain to para-hydroxybenzoate were also cloned in bacteria and yeast. To form ubiquinone, prenylated PHB undergoes several modifications such as hydroxylations, O-methylations, methylations and decarboxylation. In eukaryotes ubiquinones were found in the inner mitochondrial membrane and in other membranes such as the endoplasmic reticulum, Golgi vesicles, lysosomes and peroxisomes. Still, the subcellular site of their biosynthesis remains unclear. Considering the diversity of functions of ubiquinones, and their multistep biosynthesis, identification of factors regulating their cellular level remains an elusive task. PMID- 11051213 TI - QSARs of some novel isosteric heterocyclics with antifungal activity. AB - QSAR analysis of a set of previously synthesized 2,5,6-trisubstituted benzoxazole, benzimidazole and 2-substituted oxazolo(4,5-b)pyridine derivatives tested for growth inhibitory activity against Candida albicans, was performed by using the computer-assisted multiple regression procedure. The activity contributions for either heterocyclic ring systems or substituent effects of these compounds were determined from the correlation equation and the predictions for the lead optimization were described. The resulting QSAR revealed that the oxazolo(4,5-b)pyridine ring system with the substitution of a benzyl moiety at position 2 was the most favourable structure among the heterocyclic nuclei. Moreover, the fifth position in the fused ring system is found more significant than the other positions in improving the activity. PMID- 11051214 TI - Studies on successful aging and longevity: Polish Centenarian Program. PMID- 11051215 TI - Risk factors determining chemotherapeutic toxicity in patients with advanced colorectal cancer. AB - Antitumour therapy in advanced colorectal cancer has limited efficacy. For decades, fluorouracil has been the main anticancer drug for the treatment of colorectal cancer. Recently, however, new agents have been introduced: raltitrexed, irinotecan and oxaliplatin. Currently, the dosage for an individual patient is calculated from the estimated body surface area of the patient. Toxicity, however, frequently necessitates decreasing the dosage, extending the dose interval or even discontinuing treatment. Risk factors with predictive value for toxicity have been identified in several studies. These risk factors are often determined by the pharmacokinetic and pharmacodynamic properties of the drug. In this review, the risk factors for toxicity of the cytotoxic agents used in the treatment of advanced colorectal cancer are considered. For fluorouracil, age, gender, performance status, genetic polymorphism of dihydropyridine dehydrogenase, drug administration schedule, circadian rhythm of plasma concentrations, history of previous chemotherapy-related diarrhoea, xerostomia, low neutrophil levels, and drug-drug interactions have been identified as affecting chemotherapeutic toxicity. For raltitrexed, gender and renal and hepatic impairment, and for oxaliplatin, renal impairment and circadian rhythm of plasma concentrations, respectively, can be considered as risk factors for toxicity. In addition, age, performance status, bilirubinaemia, genetic polymorphism of uridine 5'-diphosphate-glucuronyltransferase-1A1 and drug administration schedule have been shown to be related to irinotecan toxicity. The available literature suggests that dose adjustment based on these risk factors can be used to individualise the dose in order to decrease toxicity and to improve the therapeutic index. This also applies to therapeutic drug monitoring, which has been shown to be effective controlling the toxicity of fluorouracil in some studies. Future research is warranted to assess the potential advantage of dose individualisation of chemotherapy founded on risk factors, over direct dose calculation from the estimated body surface area, with regard to toxicity, therapeutic index, and quality of life, in patients with advanced colorectal cancer. PMID- 11051217 TI - Quetiapine: a review of its safety in the management of schizophrenia. AB - Quetiapine, a dibenzothiazepine derivative, is a atypical antipsychotic which has greater in vitro binding affinity for serotonin 5-HT2 receptors than for dopamine D2 receptors. Quetiapine effectively treats both the positive and the negative symptoms of schizophrenia and is also associated with an incidence of extrapyramidal symptoms no greater than placebo across the entire dose range. In addition, it does not cause persistent hyperprolactinaemia. Quetiapine is associated with high levels of patient acceptability and satisfaction, which may result from its combination of efficacy and relatively benign adverse effect profile. The drug is well tolerated and has a low propensity to cause adverse events both during acute and long term treatment in the adult populations. The adverse effect profile of quetiapine makes the drug advantageous for patient populations who are susceptible to the adverse effects of drugs. Indeed, preliminary data show quetiapine to be very well tolerated in the elderly. Overdoses of quetiapine of up to 20g have been reported; however, with appropriate management in an intensive care setting there have been no reported fatalities. Quetiapine is metabolised by the cytochrome P450 3A4 isoenzyme, and the dose may need to be adjusted if quetiapine is co-administered with drugs which affect the activity of this isoenzyme. Overall, quetiapine has a favourable risk-benefit profile that should make it a valuable first-line agent in the treatment of schizophrenia. PMID- 11051216 TI - Drug-induced rheumatic disorders: incidence, prevention and management. AB - The purpose of this article is to review the causes, the clinical manifestations and the management of the more frequent drug-induced rheumatic disorders. These include: (i) articular and periarticular manifestations induced by fluoroquinolones, nonsteroidal anti-inflammatory drugs, injections of corticosteroids, and retinoids; (ii) multisystemic manifestations such as drug induced lupus and arthritis induced by vaccination, Bacillus Calmette-Guerin therapy and cytokines; (iii) drug-induced disorders of bone metabolism (corticosteroid-induced osteoporosis, drug-induced osteomalacia and osteonecrosis); and (iv) iatrogenic complex regional pain syndromes. Disorders caused by nonpharmacological and rarely used treatments have been deliberately excluded. Knowledge of these drug-induced clinical symptoms or syndromes allows an earlier diagnosis and treatment, and earlier drug withdrawal if necessary. With the introduction of new medications such as the recombinant cytokines and antiretroviral treatments, the number of drug-induced rheumatic disorders is likely to increase. PMID- 11051218 TI - A risk-benefit assessment of intranasal triamcinolone acetonide in allergic rhinitis. AB - The efficacy of intranasal triamcinolone acetonide in seasonal and allergic rhinitis has been evaluated in clinical trials and has been compared with antihistamines and other intranasal corticosteroids. Intranasal corticosteroids are either as equally effective as or more effective than comparative drugs. Intranasal corticosteroids are particularly useful as they decrease membrane permeability and inhibit both early and late phase reactions to allergens. They minimise the nasal secretory response and reduce the sensitivity of local nasal irritant receptors. A potential benefit of topical application is the flushing action of the nasal mucosa, which may reduce allergens and secretions. In addition to seasonal and perennial rhinitis, intranasal corticosteroids have additional benefits when used to reduce inflammation in the treatment of sinusitis and may help in decreasing secondary rhinovirus infections. Furthermore, suboptimal control of asthma can be avoided by treatment of allergic rhinitis with intranasal corticosteroids. In clinical trials, common adverse effects for triamcinolone acetonide include sneezing, dry, mucosa, nasal irritation, sinus discomfort, throat discomfort, epistaxis and headache. Posterior subcapsular cataract formation has not been seen with triamcinolone acetonide. Recent literature evaluating systemic absorption of intranasal corticosteroids have shown surprising results where significant absorption has occurred with intranasal budesonide and fluticasone propionate. Growth and hypothalamic pituitary axis (HPA) function studies have been reviewed, with some intranasal corticosteroids showing changes with continual use. A retrospective study in children receiving daily triamcinolone acetonide for 12 months showed no effect on height and bodyweight. Triamcinolone acetonide at standard dosages (110 or 220microg once or twice a day) does not appear to suppress adrenal gland function and is effective in relieving most symptoms of allergic rhinitis. The International Consensus Conference Proceedings on Rhinitis now currently recommends the use of intranasal corticosteroids as first line therapy, since they have been found to be well tolerated and effective with minimal adverse effects and, specifically, no cognitive impairment. The recommended maximum dose of aqueous triamcinolone acetonide in adults and children is 220microg once a day. The aerosol form may be recommended in children between 7 and 12 years old, up to 440microg once a day or in divided doses. Duration of allergy treatment is generally for the length of each allergy season. If symptoms are perennial, then a reduction of dosage is made to the lowest effective dose with monitoring every 3 months for risk and benefit assessment. Complications to watch for include bleeding, and possible septal perforation and nasal candidiasis, although these are rare. PMID- 11051219 TI - Minimising the risks of allergen-specific injection immunotherapy. AB - The clinical advantages of allergen-specific immunotherapy are counterbalanced by the risk of inducing systemic adverse effects. Although the frequency of life threatening systemic reactions is low, the treatment carries a risk of inducing anaphylactic reactions. A fundamental point in risk assessment is to use a clinically meaningful and internationally accepted grading system for reactions. Of importance in minimising the risk of systemic adverse effects is the identification of at-risk patients and factors, the institution of procedures for monitoring patients before injections, and the adjustment of dosages in accordance with defined rules. Asthma, especially uncontrolled asthma, is a significant risk factor for the induction of systemic reactions. Likewise, dose escalation during allergen exposure, i.e. during pollen seasons, increases the risk of adverse effects. It is recommended that standardised extracts with a documented potency and consistency between production batches are used in order to prevent overdose when changing to a new vial. The intensity of the induction regimen is a balance between the risk of inducing systemic reactions and the time required to administer the regimen. Single injections once a week are generally well tolerated, in contrast to rush immunotherapy which may carry an increased frequency of adverse effects. A clustered induction regimen (2 to 4 injections per visit) represents a compromise of a patient-friendly fast regimen without an unacceptably high frequency of systemic reactions. A major issue in improving the safety of allergen injections is minimising the human factor, e.g. mistakes of patient identification, allergen extracts and dosages. Meticulous care in monitoring every patient before the injection, which requires education and training of the staff in the dosage decision process, is the cornerstone in reducing adverse effects. Involving the patient actively in the safety monitoring process might be helpful and improves patient compliance by allowing the patient to be an active partner in the treatment. Finally, if anaphylactic reactions are induced, a successful outcome is related to the staff being able to identify the early signs and to institute immediate rescue treatment. A quality assurance programme is the optimal way to minimise the risk of immunotherapy-associated systemic reactions. PMID- 11051221 TI - Innovations in phase 1 trial design: where do we go next? PMID- 11051220 TI - Liver damage associated with minocycline use in acne: a systematic review of the published literature and pharmacovigilance data. AB - OBJECTIVE: Minocycline is an antibacterial drug used in the treatment of acne. Concern has been expressed over the possibility of severe adverse reactions to minocycline, including hepatitis. This study set out to identify and characterise reported cases of hepatotoxicity associated with the use of minocycline. METHODS: A systematic review of the literature including a search of computerised databases and analysis of data from the Uppsala Monitoring Centre (WHO Collaborating Centre for International Drug Monitoring) was conducted. The review involved a search for original case reports involving liver damage in people using minocycline. Patients taking minocycline for reasons other than acne or those given intravenous minocycline were excluded. The search strategy involved an enquiry of computerised databases and a search for secondary references. Cases were then classified appropriately. RESULTS: 65 reported cases of hepatitis or liver damage in association with minocycline from either case reports or case series were identified from the literature review. 58% of cases occurred in females and 94% were aged under 40 years. For 20 case reports there was insufficient information to classify the type of event, but for the remaining 45, 2 types of hepatic reaction were recognised: autoimmune hepatitis associated with lupus-like symptoms occurring after a median duration of exposure to minocycline of 365 days in females (n = 20) and 730 days in males (n = 9), hypersensitivity reaction associated with eosinophilia and exfoliative dermatitis occurring within 35 days of therapy (n = 16). Reports to the WHO of hepatic adverse drug reactions associated with minocycline accounted for 6% (493) of all minocycline-related adverse drug reactions (8025). The pattern of distribution in relation to exposure demonstrated 2 groups, similar to that described by the case reports. CONCLUSIONS: Severe cases of minocycline-associated hepatotoxicity appear to be a hypersensitivity reaction and occur within a few weeks of commencing therapy. An autoimmune hepatitis usually presents after exposure to minocycline of a year or more, is more common in women and is sometimes associated with lupus-like symptoms. PMID- 11051222 TI - Molecular assays for the diagnosis of minimal residual head-and-neck cancer: methods, reliability, pitfalls, and solutions. AB - The prognosis of cancer patients is determined by the radicalness of treatment: residual tumor cells will grow out and develop in manifest local recurrences, regional recurrences, and distant metastases. Classical diagnostic methods such as radiology and histopathology have limited sensitivities, and only by molecular techniques can minimal residual disease be detected. In tissue samples containing the normal tissue counterpart of a tumor, only tumor-specific markers can be exploited, whereas in other samples, tissue-specific markers can be used. At present, there are two main methodologies in use, one based on antigen-antibody interaction and the other based on amplified nucleic acids. The most commonly used nucleic acid markers are mutations or alterations in tumor DNA (tumor specific markers) or differentially expressed mRNA (tissue-specific markers). Many reports and reviews have been published on the assessment of minimal residual disease by molecular markers, showing either positive or negative clinical correlations. One of the main reasons for these contradictory findings is the technical difficulty in finding the small numbers of tumor cells in the large number of normal cells, which necessitates sensitivities of the assays up to 1 tumor cell in 2 x 10(7) normal cells. These assays often are complex, demand considerable experience, and usually are laborious. In this review, we will address a number of the technical issues related to molecular assays for tumor cell detection that make use of nucleic acids as markers. Many difficulties in data interpretation are at least in part because of technical details that might have been solved by the incorporation of one or more appropriate controls. We hope that this review clarifies a number of these issues and help clinicians and investigators interested in this field to understand and weigh the contradictory findings in the published studies. This will help move the field forward and facilitate clinical implementation. PMID- 11051223 TI - Detection of circulating cancer cells with von hippel-lindau gene mutation in peripheral blood of patients with renal cell carcinoma. AB - Mutations of the von Hippel-Lindau (VHL) tumor suppressor gene have been detected in up to 60% of sporadic clear cell renal carcinomas (RCCs). Even patients with RCCs believed to be curable with radical nephrectomy sometimes develop distant metastasis 5-10 years after surgery, suggesting hematogenous circulation of cancer cells. Useful tumor markers have not yet been established for RCC. To detect patients at high risk of metastasis after surgery, we developed a highly sensitive and specific nested reverse transcription-PCR method using VHL gene mutation to detect circulating cancer cells. We screened 29 sporadic clear cell RCCs from patients for mutations of the VHL gene by direct sequencing. We next examined blood samples from patients with the VHL gene mutation using mutation specific nested reverse transcription-PCR. Somatic mutations were detected in 20 of 29 (69.0%) sporadic clear cell RCCs. The VHL gene mutations were detected in peripheral and/or renal venous blood from 15 of 20 (75%) patients. The mutations were detected in the peripheral blood in 2 of 17 (11.8%) patients before surgery, 6 of 16 (37.5%) patients within 24 h after surgery, 3 of 16 (18.8%) patients on day 7 after surgery, and 2 of 11 (18.2%) patients on day 30 after surgery. In seven of nine (77.8%) patients, mutations were detected in renal venous blood during surgery. These findings indicate the presence of circulating cancer cells with VHL gene mutation. Although much larger studies are needed to determine the clinical significance, our study shows that this technique is feasible for detecting circulating RCC cells. PMID- 11051224 TI - Telomerase RNA as a detection marker in the serum of breast cancer patients. AB - Tumor-derived circulating DNA has been found in the plasma of cancer patients. Alterations include decreased strand stability, mutations of oncogenes or of tumor suppressor genes, microsatellite alterations, and hypermethylation of several genes. RNA has also been found circulating in the plasma of normal subjects and cancer patients. Tyrosinase mRNA has been extracted from the serum of melanoma patients and subjected to RT-PCR. Moreover, the presence of cell-free EBV-associated RNA has been reported in the plasma of patients with nasopharyngeal carcinoma. Human telomerase comprises two RNA subunits, telomerase RNA template (hTR) and its catalytic component, telomerase reverse transcriptase protein (hTERT). Expression of these subunits correlates with telomerase activity. Using RT-PCR, we investigated whether these RNA subunits were present in the serum of 18 patients with breast cancer, 2 patients with benign breast disease, and 21 normal subjects. The presence of amplifiable RNA was confirmed in all tissue and serum samples using RT-PCR of glyceraldehyde-3-phosphate dehydrogenase RNA. hTR was found in 17 of 18 tumors (94%) and 5 of 18 serum samples (28%). hTERT was also detected in 17 of 18 tumors (94%) and in 4 of 16 available serum samples (25%). hTR and hTERT were undetectable in tissues and sera taken from 2 patients with benign disease and in the sera of 21 normal subjects. We conclude that RNA is detectable in the serum of breast cancer patients and that tumor-derived mRNA can be extracted and amplified using RT-PCR, even in patients with localized disease. This may have implications for cancer diagnosis and follow-up in the future. PMID- 11051225 TI - Antisense inhibition of hMLH1 is not sufficient for loss of DNA mismatch repair function in the HCT116+chromosome 3 cell line. AB - We have reported that transfer of chromosome 3 (Chr3) containing a single wild type copy of the hMLH1 gene into HCT116 colon cancer cells, a cell line deficient in DNA mismatch repair (MMR) activity attributable to inactivating hMLH1 mutations, corrects all of the aspects of the MMR repair-deficient phenotype. We inhibited the expression of the wild-type hMLH1 gene using antisense RNA in HCT116+Chr3 cells to determine if this would result in reversion to the MMR deficient phenotype. Despite profound inhibition of hMLH1 expression, DNA MMR activity and alkylation sensitivity were not impaired in the antisense transfected HCT116+Chr3 cells. Additionally, arrest of the cell cycle at the G2 phase with alkylation damage occurs in these cells, a phenotype associated with MMR proficiency. These results indicate that even with a reduction in the expression of hMLH1 protein below the limits of detection by Western blotting, DNA MMR activity remained fully functional (by direct DNA MMR activity assay). We would speculate that hMLH1 is expressed in substantially greater abundance than would be minimally necessary for DNA MMR and that minor reductions in the expression of this protein would not be sufficient to permit DNA MMR dysfunction. Alternatively, Chr3 may contain a second hMLH1 homologue that might overlap with the function of hMLH1. PMID- 11051226 TI - Adenovirus 5 early region 1A does not induce expression of the ewing sarcoma fusion product EWS-FLI1 in breast and ovarian cancer cell lines. AB - The adenovirus 5 early region 1A (E1A) can function as a tumor suppressor gene and is being used in clinical trials as a therapeutic agent for advanced breast, ovarian, and head and neck cancer. Recently, there has been a dispute regarding whether transfection with the E1A gene can induce expression of the Ewing sarcoma oncogenic fusion transcript EWS-FLI1 (Sanchez-Prieto et al., Nat. Med., 5: 1076 1079, 1999; Melot and Delattre, Nat. Med., 5: 1331, 1999; Kovar et al., Cancer Res., 60: 1557-1560, 2000). In an effort to settle the controversy, we tested several stable E1A transfectants of cell lines MDA-MB-231, MCF-7, MDA-MB-435 (breast cancer), SKOV3-ipl (ovarian cancer), and PC-3 (prostate cancer), as well as parental and vector-transfected controls, HEK 293 cells, and RD-ES (Ewing sarcoma) cells, for the EWS-FLI1 fusion product. The EWS-FLI1 transcript could not be identified with reverse transcription-PCR in any of the 13 E1A-transfected cell lines analyzed. Furthermore, the EWS-FLI1 fusion protein could not be detected by Western blot analysis in E1A-transfected cell lines. These results suggest that E1A transfection does not necessarily lead to expression of the oncogenic EWS-FLI1 fusion transcript. Thus, the potential induction of this gene rearrangement by E1A gene therapy is unlikely to be clinically significant in the treatment of advanced malignant disease. PMID- 11051227 TI - Lung cancer proliferation correlates with [F-18]fluorodeoxyglucose uptake by positron emission tomography. AB - Tumor proliferation has prognostic value in resected early-stage non-small cell lung cancer (NSCLC). We evaluated whether [F-18]fluorodeoxyglucose (FDG) uptake of NSCLC correlates with tumor proliferation and, thus, could noninvasively grade NSCLCs (refining patient prognosis and therapy). Thirty-nine patients with potentially resectable NSCLC underwent whole-body FDG positron emission tomography (PET) 45 min after i.v. injection of 10 mCi of FDG. Tumor FDG uptake was quantitated with the maximum pixel standardized uptake value (maxSUV). The lesion diameter from computed tomography was used to correct the maxSUV for partial volume effects using recovery coefficients determined for the General Electric Advance PET scanner. Thirty-eight patients underwent complete surgical staging (bronchoscopy and mediastinoscopy, with or without thoracotomy). One stage IV patient by PET underwent bronchoscopic biopsy only. Immunohistochemistry for Ki-67 (proliferation index marker) was performed on all of the 39 NSCLC specimens (35 resections, 1 percutaneous, and 3 surgical biopsies). The specimens were reviewed for cellular differentiation (poor, moderate, well) and tumor type. Lesions ranged from 0.7 to 6.1 cm. The correlation found between uncorrected maxSUV and lesion size (Rho, 0.56; P = 0.0006) disappeared when applying the recovery coefficients (Rho, -0.035; P = 0.83). Ki-67 expression (percentage of positive cells) correlated strongly with FDG uptake (corrected maxSUV: Rho, 0.73; P < 0.0001). The correlation was stronger for stage I lesions (11 stage IA, 15 stage IB): Rho, 0.79; P < 0.0001) and strongest in stage IB (Rho, 0.83; P = 0.0019). A significant association (P < 0.0001) between tumor differentiation and corrected SUV was noted. FDG PET may be used to noninvasively assess NSCLC proliferation in vivo, identifying rapidly growing NSCLCs with poor prognosis that could benefit from preoperative chemotherapy. PMID- 11051228 TI - Effect of 13-cis-retinoic acid on serum prostate-specific antigen levels in patients with recurrent prostate cancer after radical prostatectomy. AB - The objective of this study was to determine whether there is any beneficial effect of oral 13-cis-retinoic acid (isotretinoin) on prostate cancer, using serum prostate-specific antigen (PSA) levels as a surrogate end point in patients with a rising serum PSA after radical prostatectomy. In the first phase, the effect of the drug on the serum PSA level was tested in 14 control patients with normal prostates. Our goal was to exclude any effect of isotretinoin on PSA secretion and metabolism and thus to validate any observed effect on PSA as indicative of anticancer activity. In the second phase, patients with rising PSA levels after radical prostatectomy and no evidence of metastatic disease were treated with oral isotretinoin at a daily dose of 1.0 mg/kg. Serum PSA levels were checked monthly for the first 4 months after initiation of treatment and every 3 months thereafter. No significant changes in serum PSA levels after 3 months of isotretinoin treatment were recorded in the control group (P = 1.000). Three of 11 postprostatectomy patients (27%) had a PSA reduction of 28%, 15%, and 6.6% after initiation of treatment that lasted for a period of 2-3 months. In two of these three patients, the PSA levels subsequently rose exponentially. Another patient displayed a stabilization of the serum PSA curve for 3 months after an initial sharp rise. No grade 3 or 4 toxicity was recorded in this group of patients. Isotretinoin had a modest, transient effect on the serum PSA level in 4 of 11 (36%) patients with a rising serum PSA after radical prostatectomy. We conclude that this drug is unlikely to be of major therapeutic benefit in prostate cancer patients when used as a single agent. However, its modest effect argues for the exploration of other, more potent retinoids for prostate cancer therapy. PMID- 11051229 TI - Bioavailability study of oral liquid and tablet forms of alpha difluoromethylornithine. AB - The purpose of this study was to assess the bioavailability of two oral preparations of difluoromethylornithine (DFMO). The current preparation of DFMO is a liquid with a concentration of 0.2 gram/ml that must be drawn up into a syringe and dispensed into a small medicine glass. This form of DFMO causes wastage of the medication. The liquid form also makes compliance and blinding difficult. Recently, a new coated tablet preparation has become available from Ilex Oncology Services (San Antonio, TX). The coated tablets are 0.25 gram and are scored. The tablet form should increase compliance by making it much easier for the subject to take the medication. This report compares the bioavailability of both preparations with the goal of demonstrating equivalence of the preparations. Ten normal subjects entered the cross-over study in which the order in which they would receive the liquid or tablet preparation of DFMO was randomized. The study was designed with the objective of establishing the bioequivalence of a tablet preparation of DFMO at daily dose 0.5 gram/m2 and a liquid preparation of DFMO at the same daily dose. The mean area under the time by-concentration curves (microM x hours) for the liquid and tablet preparations was 368.2 and 370.4, respectively. The peak concentrations for the liquid and tablet preparations were 47.3 and 48.2 microM, respectively. No statistically significant differences were seen in these parameters, in time to peak concentration, or in serum half-life. PMID- 11051230 TI - A phase I radioimmunotherapy trial evaluating 90yttrium-labeled anti carcinoembryonic antigen (CEA) chimeric T84.66 in patients with metastatic CEA producing malignancies. AB - Chimeric T84.66 (cT84.66) is a genetically engineered human/murine chimeric IgG, with high affinity and specificity to carcinoembryonic antigen (CEA). The purpose of this Phase I dose escalation therapy trial was to evaluate the toxicities, biodistribution, pharmacokinetics, tumor targeting, immunogenicity, and organ and tumor absorbed dose estimates of cT84.66 labeled with 90Y. Patients with metastatic CEA-producing malignancies were first administered 5 mCi 111In-labeled DTPA-cT84.66 (5 mg), followed by administration of the therapy dose of 90Y labeled DTPA-cT84.66 1 week later. The therapy infusion was immediately followed by a 72-h administration of DTPA at 250 mg/m2/24 h. Dose levels of administered activity ranged from 5 to 22 mCi/m2 with three to six patients per level. Serial nuclear scans, blood samples, and 24-h urine collections were performed out to 5 days after infusion. Human antichimeric antibody response was assayed out to 6 months. Patients were administered up to 3 cycles of therapy every 6 weeks. Radiation absorbed doses to organs were estimated using a five compartment model and MIRDOSE3. Twenty-two patients received at least one cycle of therapy, with one individual receiving two cycles and two receiving three cycles of therapy. All were heavily pretreated and had progressive disease prior to entry in this trial. Reversible leukopenia and thrombocytopenia were the primary dose-limiting toxicities observed. Maximum tolerated dose was reached at 22 mCi/ m2. In general, patients with liver metastases demonstrated more rapid blood clearance of the antibody. Thirteen patients developed an immune response to the antibody. Average radiation doses to marrow, liver, and whole body were 2.6, 29, and 1.9 cGy/mCi 90Y, respectively. Dose estimates to tumor ranged from 66 to 1670 cGy (8.7 to 52.2 cGy/mCi 90Y) for each cycle of therapy delivered. Although no major responses were observed, three patients demonstrated stable disease of 12-28 weeks duration and two demonstrated a mixed response. In addition, a 41-100% reduction in tumor size was observed with five tumor lesions. 90Y-labeled cT84.66 was well tolerated, with reversible thrombocytopenia and leukopenia being dose limiting. Patients with extensive hepatic involvement by tumor demonstrated unfavorable biodistribution for therapy with rapid blood clearance and poor tumor targeting. Average tumor doses when compared with red marrow doses indicated a favorable therapeutic ratio. Stable disease and mixed responses were observed in this heavily pretreated population with progressive disease. This trial represents an important step toward further improving the therapeutic potential of this agent through refinements in the characteristics of the antibody and the treatment strategies used. Future trials will focus on the use of peripheral stem cell support to allow for higher administered activities and the use of combined modality strategies with radiation-enhancing chemotherapy drugs. Further efforts to reduce immunogenicity through humanization of the antibody are also planned. Finally, novel engineered, lower molecular weight, faster clearing constructs derived from cT84.66 continue to be evaluated in preclinical models as potential agents for radioimmunotherapy. PMID- 11051231 TI - Green tea polyphenol treatment to human skin prevents formation of ultraviolet light B-induced pyrimidine dimers in DNA. AB - Cancer chemopreventive effects of polyphenols from green tea (GTP) in mouse models of photocarcinogenesis are established. The present study is extended from mouse model to human system in vivo to determine the effect of topical application of GTP to human individuals against UV light-induced DNA damage in the form of cyclobutane pyrimidine dimers (CPDs) in the skin. UVB-induced CPDs were detected by immunohistochemical technique using monoclonal antibodies to thymine dimers. With the gradual increase in UVB dose, both erythema response and CPD formation in the skin was increased. GTP treatment inhibited both UVB-induced erythema response as well as CPD formation. Topical treatment with GTP (approximately 1 mg/cm2 of skin area) 20 min before human buttock skin (sun protected site) exposure to UVB inhibited CPD formation in epidermis by 81, 70, 60, and 60% at 0.5, 1.0, 2.0, and 4.0 minimal erythema dose of UV exposure, respectively. Treatment of human skin with varying doses of GTP (1-4 mg/2.5 cm2 of skin area) before a single dose of UVB exposure (4.0 minimal erythema dose) decreased dose dependently the formation of UVB-induced CPDs in both epidermis and dermis. The inhibition of UVB-induced CPDs by GTP treatment may be, at least in part, responsible for the inhibition of photocarcinogenesis. Our data suggest that GTP may be used as a novel chemopreventive candidate and possible strategy to reduce UV-induced skin cancer risk in the human population. PMID- 11051232 TI - Chronic daily low dose of 4-methyl-5-(2-pyrazinyl)-1,2-dithiole-3-thione (Oltipraz) in patients with previously resected colon polyps and first degree female relatives of breast cancer patients. AB - The chemoprevention agent oltipraz, one of the most active chemopreventive compounds in preclinical studies, has been shown to induce glutathione-S transferase (GST) activity in animals. Oltipraz was evaluated in a Phase I trial at daily oral doses of 20 mg (L1), 50 mg (L2), and 100 mg (L3) and twice weekly doses of 125 mg (L4) taken for 6 months with 6 patients entered at L1 and L2 and 7 patients entered at L3 and L4 (26 subjects: 19 females and 7 males). The subject population included patients with previously resected colon polyps and first-degree female relatives of breast cancer patients. Patients with resected colon polyps underwent rectal biopsy for GST and glutathione (GSH) analyses. Of the 26 subjects, the following completed 6 months of therapy: 4 of 6 patients (L1), 4 of 6 patients (L2), 5 of 7 patients (L3), and 4 of 7 patients (L4). Toxicities were mild to severe and included: gastrointestinal symptoms, photosensitivity/heat intolerance, and neurological symptoms. Monthly plasma samples were obtained 2-3 h after oltipraz ingestion with minimally detectable plasma concentrations at L1. There was a significant difference in mean oltipraz concentration across the four doses, with no significant differences in mean oltipraz concentration over time. Rectal tissue and lymphocyte GSH and GST were variable, with no significant difference in mean levels across doses. At the 100 mg/day dose (L3), 1 patient experienced significant increase in rectal tissue GSH and GST activity, whereas 3 additional patients (L1 and L4) had >50% increase in tissue GSH. Lymphocyte GSH level was significantly related to plasma oltipraz concentration. There were no significant correlations between plasma oltipraz concentration and lymphocyte GST level nor any significant correlation between plasma concentration and percentage of change in tissue GSH or GST. Further investigation of dose/schedule and biological end points is ongoing. PMID- 11051233 TI - Phase III randomized study of postradiotherapy chemotherapy with alpha difluoromethylornithine-procarbazine, N-(2-chloroethyl)-N'-cyclohexyl-N nitrosurea, vincristine (DFMO-PCV) versus PCV for glioblastoma multiforme. AB - Although the efficacy of the nitrosourea-based combination chemotherapy procarbazine, N-(2-chloroethyl)-N'-cyclohexyl-N-nitrosurea, and vincristine (PCV) has been previously demonstrated in the setting of anaplastic/intermediate-grade gliomas, the benefit for glioblastoma patients remains unproven. In the current study, we sought to determine whether the addition of alpha difluoromethylornithine (eflornithine), an inhibitor of ornithine decarboxylase, which has shown encouraging results in the setting of recurrent glioma patients, to a nitrosourea-based therapy (PCV) would constitute a more effective adjuvant therapy in the treatment of glioblastoma multiforme patients in the postradiation therapy setting. Following conventional radiation therapy, 272 glioblastoma (GBM) patients were randomized to receive either alpha-difluoromethylornithine-PCV (DFMO-PCV; 134 patients) or PCV alone (138 patients), with survival and time to tumor progression being the primary endpoints. The starting dosage of DFMO was 3.0 g/m2 p.o. q8h for 14 days before and after treatment with N-(2-chloroethyl)-N cyclohexyl-N-nitrosurea; PCV was administered as previously described1. Clinical and radiological (Gadolinium-enhanced MRI) follow-ups were nominally at the end of each 6 or 8 week cycle (PCV at 6 weeks; DFMO-PCV at 8 weeks). Laboratory evaluations for hematologic and other adverse effects were at 2 week intervals. There was no difference in median survival or median time-to-tumor progression between the two treatment groups, as measured from day of commencement of postradiotherapy chemotherapy [MS (months): DFMO-PCV, 10.5; Overall survival, as measured from time of tumor diagnosis at first surgery, was 13.3 and 14.2 months at the median and 6.2 and 8.7% at 5 years, respectively, for the DFMO-PCV and PCV arms. The treatment effect was unchanged after adjustment for age, performance status (KPS), extent of surgery, and other factors using the multivariate Cox proportional hazard model. Adverse effects associated with DFMO consisted of gastrointestinal (diarrhea nausea/vomiting), cytopenias, and minimal ototoxicity (limited to tinnitus) at the dose range tested. The addition of DFMO to the nitrosourea-based PCV regimen in this phase III study demonstrated no additional benefit in glioblastoma patients, underscoring the resistance of glioblastoma multiforme tumors to alkylating agents. For patients with anaplastic (intermediate grade) gliomas, in which the previously demonstrated benefit of post-radiation chemotherapy is more substantial, the evaluation of DFMO-PCV vs. PCV is still ongoing and hopefully will yield more encouraging results. PMID- 11051234 TI - A phase I and pharmacologic evaluation of the DNA intercalator CI-958 in patients with advanced solid tumors. AB - 5-[(2-Aminoethyl)amino]-2-[2-(diethylamino)ethyl]-2H-[1]benzothiopyra no[4,3,2 cd]-indazol-8-ol trihydrochloride (CI-958) is the most active member of a new class of DNA intercalating compounds, the benzothiopyranoindazoles. Because of its broad spectrum and high degree of activity as well as a favorable toxicity profile in preclinical models, CI-958 was chosen for further development. The Phase I study described here was undertaken to determine the toxicity profile, maximum tolerated dose, and pharmacokinetics of CI-958 given as an i.v. infusion every 21 days. Adult patients with advanced refractory solid tumors who had adequate renal, hepatic, and hematological function, life expectancy, and performance status were eligible for this study. Written informed consent was obtained from all patients. Patients received a 1- or 2-h infusion of CI-958 at 21-day intervals. The starting dose was 5.2 mg/m2, and at least three patients were evaluated at each dose level before proceeding to a new dose level. A pharmacokinetically guided dose escalation design was used until reaching a predetermined target area under the plasma concentration versus time curve (AUC), after which a modified Fibonacci scheme was used. Forty-four patients (21 men and 23 women; median age, 59 years) received 162 courses of CI-958. Neutropenia and hepatorenal toxicity were the dose-limiting toxicities, which defined the maximum tolerated dose of CI-958 to be 875 mg/m2 when given as a 2-h infusion every 21 days. There were no tumor responses. Two patients had stable disease for >250 days. The recommended Phase II dose is 560 mg/m2 for patients with significant prior chemotherapy and 700 mg/m2 for patients with minimal prior chemotherapy. Pharmacokinetic analysis of plasma and urine concentration-time data from each patient was performed. At the recommended Phase II dose of 700 mg/m2, mean CI-958 clearance was 370 ml/min/m2, mean AUC was 33800 ng-h/ml, and mean terminal half life (t1/2) was 15.5 days. The clearance was similar at all doses, and plasma CI 958 AUC increased proportionally with dose, consistent with linear pharmacokinetics. The percentage reduction in absolute neutrophil count from baseline was well predicted by AUC using a simple Emax model. The pharmacokinetically guided dose escalation saved five to six dose levels in reaching the maximum tolerated dose compared with a standard dose escalation scheme. This may represent the most successful application to date of this dose escalation technique. PMID- 11051235 TI - Unexpected cytokines in serum of malignant melanoma patients during sequential biochemotherapy. AB - Biochemotherapy, which combines traditional chemotherapy with immune modulating biologicals, produces an unexpectedly high response rate (>50%) in advanced melanoma patients. We hypothesize that immunological mechanism(s) are responsible for the increased response rate, and particularly that macrophage activation is involved in tumor reduction. Patients were randomized to receive chemotherapy, composed of cisplatin, vinblastine, and dacarbazine (CVD), or biochemotherapy, which is CVD followed by interleukin (IL)-2 and IFN-alpha2b (CVD-BIO). Laboratory analysis was performed on sera from 41 patients from each arm. Measurements of macrophage activation (neopterin), nitric oxide production (nitrite), and tumor necrosis factor-alpha (TNF-alpha), IL-1alpha, IL-1beta, IFN-gamma, IL-6, IL-10, and soluble IL-2 receptor (sIL-2R) were performed. Six of the nine biological responses (nitrite, neopterin, IFN-gamma, IL-6, soluble IL-2R, and IL-10) significantly (P < 0.0002) increased in the CVD-BIO patients but not in the CVD patients. The increased IL-6 (P = 0.04) and IL-10 (P = 0.05) correlated with patient response, but only when the minor responders were included in the analysis. Evidence of macrophage activation was found in CVD-BIO patients and not in those receiving CVD alone. In addition, an unusual cytokine elaboration composed of IL-6, IFN-gamma, IL-10, nitrite, neopterin, and sIL-2R, but not the expected TNF-alpha and IL-1, was detected. A trend of higher increase in IL-6 and IL-10 in patients having clinical response was found, suggesting an incomplete Th2 pattern of cytokine elaboration. These data show that macrophage activation does not appear to be critical in the response to CVD-BIO, but that IL-10 and IL 6 induced by the BIO component of the CVD-BIO were associated with tumor regression, and that their biology should be pursued further in the analysis of mechanism(s) of response. PMID- 11051236 TI - Loss of B7.2 (CD86) and intracellular adhesion molecule 1 (CD54) expression is associated with decreased tumor-infiltrating T lymphocytes in diffuse B-cell large-cell lymphoma. AB - Tumor-infiltrating CD8+ T-lymphocytes (T-TILs) are thought to be relevant to immunosurveillance of several tumor types including B-cell non-Hodgkin's lymphoma. B- and T-lymphocyte interactions via cellular adhesion molecules (CAMs), recognition molecules (HLAs), and costimulatory molecules (CSMs) are necessary for optimal antigen-specific T-cell activation to occur and may be important in generating effective host T-TIL responses. We previously found that low T-TIL response (CD8+ T cells < 6%) correlates with statistically shorter relapse-free survival in patients with diffuse large-cell lymphoma (DLCL). We now extend our observations in 71 DLCL patients by analyzing malignant B-cell expression of the following molecules important in T-cell activation: (a) recognition molecules [MHC I (MAS and MCA) and MHC II (HLA-DR, -DP, -DQ)]; (b) CAMs [leukocyte function antigen 1 (CD11a and CD18) and intracellular adhesion molecule 1 (CD54)]; and (c) CSMs [B7.1 (CD80) and B7.2 (CD86)]. Eighteen patients (25%) had low a T-TIL response, and 53 patients (75%) had a high T-TIL response. Overall, expression of the MHC class H molecules HLA-DR and HLA-DQ was most conserved. The loss of B7.2 (P = 0.04), intracellular adhesion molecule 1 (P = 0.0004), MAS (P = 0.02), and HLA-DR (P = 0.0004) expression was significantly associated with decreased T-TIL response. In 100% of patients with low T-TIL responses, at least one HLA, CAM, or CSM was undetectable on the malignant B cells by immunohistochemical staining (mean number of molecules lost = 2.67). In contrast, 49% of patients with high T-TIL responses had no losses in HLA, CAM, or CSM expression (mean number of molecules lost = 0.89). The mean number of absent molecules (HLA, CAM, or CSM) was significantly associated with T-TIL response (P = 0.0001). We conclude that loss of HLA, CAM, or CSM expression on malignant B cells is associated with a poor host T-cell immune response. In addition, because patients with low T-TIL response had lost expression of multiple cellular adhesion, recognition, and costimulatory molecules, our results suggest that a combination of immunorestorative therapies may be required to generate effective antitumor T-cell responses in B-cell DLCL. PMID- 11051237 TI - P73 gene expression in ovarian cancer tissues and cell lines. AB - Thep73 gene, a homology of p53, is a new candidate of imprinting and tumor suppressor gene. To investigate the role of p73 in ovarian cancer, we studied the allelic expression in 56 cases of ovarian cancer using StyI polymorphism analysis. We also examined p73 expression by semi-quantitative reverse transcription-PCR as well as by Western blot analysis and DNA methylation study of the CpG island in exon 1 in ovarian cancer tissues and cell lines. Loss of heterozygosity was found in 8.3% (2 of 24) of the cases. Biallelic expression was demonstrated in 91.7% (22 of 24) of the tumor samples, in 70.8% (17 of 24) of the normal samples, and in 1 ovarian cancer cell line. Imbalanced expression and monoallelic expression were found in three and two pairs of matched samples, respectively. Overexpression of p73 was found in advanced ovarian cancer rather than in early-stage disease or in borderline ovarian tumor. No significant difference was found in the p53 expression. Three cell lines with absent p73 protein expression and one tumor sample with monoallelic expression were methylated in the CpG island. Demethylation in SKOV3 cell line using 5 azacytidine can reactivate the expression of this gene in both the mRNA and the protein level. Our results indicated that p73 was not imprinted in most of the ovarian cancer and normal tissues, but it could be involved in the advanced ovarian cancer through overexpression. DNA methylation may contribute to the lack of p73 expression. PMID- 11051238 TI - Expression of cancer testis genes in human brain tumors. AB - Cancer-testis (CT) genes are expressed in a variety of human cancers but not in normal tissues, except for testis tissue, and represent promising targets for immunotherapeutic and gene therapeutic approaches. Because little is known about their composite expression in human brain tumors, we investigated the expression of seven CT genes (MAGE-3, NY-ESO-1, HOM-MEL-40/SSX-2, SSX-1, SSX-4,HOM-TES 14/SCP-1, and HOM-TES-85) in 88 human brain tumor specimens. Meningiomas expressed only HOM-TES-14/SCP-1 (18% of meningiomas were HOM-TES-14/SCP-1 positive) and did not express any other CT genes. One ependymoma was negative for all CT genes tested. SSX-4 was the only CT gene expressed in oligodendrogliomas (2 of 5 cases), and it was also expressed in oligoastrocytomas (3 of 4 cases) and astrocytomas (10 of 37 cases). Astrocytomas were most frequently positive for HOM TES-14/SCP-1 (40%) and SSX-4 (27%), followed by HOM-TES-85 (13%), SSX-2 (11%), and MAGE-3 (7%). Whereas MAGE-3 was detected only in grade IV astrocytomas, the expression of the other CT genes showed no clear correlation with histological grade. Of 39 astrocytomas, 60% expressed at least one CT gene, 21% expressed two CT genes, and 8% coexpressed three CT genes of the seven CT genes investigated. We conclude that a majority of oligoastrocytomas and astrocytomas might be amenable to specific immunotherapeutic interventions. However, the identification of additional tu-mor-specific antigens with a frequent expression in gliomas is warranted to allow for the development of widely applicable polyvalent glioma vaccines. PMID- 11051239 TI - Heterogeneity in the clinical phenotype of TP53 mutations in breast cancer patients. AB - TP53 mutation is a strong independent marker for survival in breast cancer with some heterogeneity in the clinical phenotype of various types of mutations. Based on 315 patients with breast carcinoma, we suggest a new model for the differentiation of TP53 mutations. Although TP53 mutation in general was associated with aggressive tumor/patient characteristics, missense mutations outside any conserved or structural domain did not affect the clinical outcome (risk of disseminated disease and death). In contrast, patients with missense mutations affecting amino acids directly involved in DNA or zinc binding displayed a very aggressive clinical phenotype. Null mutations (including missense mutations disrupting the tetramerization domain) and the remaining missense mutations displayed an intermediate aggressive clinical phenotype. When patients with primary early breast cancer were divided into three groups (wild type together with missense mutations outside structural/conserved domains, null mutations and missense mutations with intermediate clinical phenotype, and very aggressive missense mutations), disease-specific survival rates were 89%, 58%, and 35% (5-year actuarial values, P < 0.0001), respectively. In a Cox proportional hazards analysis, separation of TP53 mutations according to these criteria eliminated the prognostic importance of all investigated classical factors except nodal status. PMID- 11051240 TI - Contrasted frequencies of p53 accumulation in the two age groups of North African nasopharyngeal carcinomas. AB - EBV-associated nasopharyngeal carcinomas (NPCs) from Southeast Asia and North Africa have many common clinical and biological characteristics. However, they differ with regard to their age distribution. In Asia, NPC mainly affects patients in the 4th or 5th decade of their life, whereas in North Africa an additional peak of incidence is found between the ages of 10 and 20. The p53 gene is rarely mutated in NPC. However, several groups have reported a consistent accumulation of p53 in Asian NPCs. To determine whether p53 was also accumulated in North African NPCs, we investigated its expression, by immunohistochemistry, in a series of 90 Tunisian biopsies. Bc12 and CD95, two proteins involved in the regulation of cell survival and apoptosis, were investigated in the same study. We found accumulation of p53 in 81% of the cases for patients over 30 years of age, but in only 38% of specimens for younger patients (P = 0.00013). There was a trend toward a higher frequency of Bc12 detection in patients over 30, but it was not statistically significant. CD95 expression was detected in all biopsies, generally at a high level, even at advanced stages of the disease. The changing frequency of p53 accumulation, below and over 30, suggests that NPC cells often achieve malignant transformation through different pathways in both age groups. PMID- 11051241 TI - Functional evaluation of p53 and PTEN gene mutations in gliomas. AB - We screened mutations of two major tumor suppressor genes, p53 and PTEN, in 66 human brain tumors using a yeast-based functional assay and cDNA-based direct sequencing, respectively. The frequency of p53 mutations was 28.8% (19 of 66) and was higher in anaplastic astrocytoma (9 of 14, 64.3%,) than in glioblastoma multiforme (GBM; 7 of 27, 25.9%,), supporting previous speculation that there are at least two genetic pathways leading to GBM, a de novo pathway without p53 mutation and a "progressive" pathway with p53 mutation. PTEN mutation was observed in 8 of 64 tumors (12.5%), mainly GBMs (7 of 26, 26.9%), both with and without p53 mutation. These results suggest that mutation of the PTEN gene is a later event than that of the p53 gene in glioma progression and is associated with both the genetic pathways. All of the detected PTEN missense mutations and an in-frame small deletion inactivated PTEN phosphoinositide phosphatase activity in vitro. Because the tumors containing PTEN mutations also showed loss of heterozygosity in the chromosome 10q23 region flanking the PTEN gene, our data clearly indicate that inactivation of both PTEN alleles occurs in a subset of high-grade gliomas, therefore confirming the previous idea that PTEN acts as a tumor suppressor gene. PMID- 11051242 TI - Overexpression of matrix metalloproteinase 2 predicts unfavorable outcome in early-stage non-small cell lung cancer. AB - This prospective study was performed to assess the impact of matrix metalloproteinase (MMP) 2 expression on the clinical course of patients with operable non-small cell lung cancer (NSCLC). Specimens of 193 consecutive patients with completely resected NSCLC were examined for MMP-2 expression by immunohistochemical staining with a polyclonal antibody. Homogeneous immunostaining of cancer cells was considered positive and heterogeneous, or no staining was considered negative concerning overexpression of MMP-2. Four specimens were excluded from further analyses because of unspecific staining. The median follow-up period was 71.5 months (range, 12-120 months). Overexpression of MMP-2 was observed in 64 (33.9%) of 189 patients and did not correlate with clinicopathoiogical parameters. In patients without lymph node involvement (pN0 stage) MMP-2 overexpression was an independent prognostic parameter for unfavorable outcome: Log-rank analysis showed a significant association of MMP-2 overexpression with shortened cancer-related survival (P = 0.04) and disease-free survival (P = 0.03). Multivariate regression analysis confirmed MMP-2 overexpression as predictor of shortened cancer-related survival in NSCLC without lymph node involvement (P = 0.005, relative risk, 2.6). The present study revealed that MMP-2 overexpression predicts a poor prognosis in early-stage NSCLC. Therefore, it might be worth investigating the role of MMP inhibitors as adjuvant therapeutic agents in NSCLC. PMID- 11051243 TI - Identification of beta-tubulin isoforms as tumor antigens in neuroblastoma. AB - There is currently substantial interest in the identification of human tumor antigens for diagnosis and immunotherapy of cancer. We have implemented a proteomic approach for the identification of tumor proteins that elicit a humoral response in cancer patients, which we have applied to neuroblastoma. Proteins from neuroblastoma tumors and cell lines were separated by two-dimensional PAGE and transferred to poly(vinylidene difluoride) membranes. Sera from 23 newly diagnosed patients with neuroblastoma, from 12 newly diagnosed children with other solid tumors, and from 13 normal individuals were screened for IgG and IgM autoantibodies against neuroblastoma proteins by means of Western blot analysis. Sera from 11 patients with neuroblastoma and from 1 patient with a primitive neuroectodermal tumor, but none of the other controls exhibited IgG-based reactivity against a protein constellation with an estimated Mr 50,000. NH2 terminal sequence and mass spectrometric analysis identified the major constituents of this constellation as beta-tubulin isoforms I and III. The IgG antibodies were additionally characterized to be of the subclass IgG1. Neuroblastoma patient sera that contained anti-beta-tubulin IgG antibodies also contained IgM antibodies specific against the full-length beta-tubulin molecule and against COOH-terminal beta-tubulin cleavage products. Neuroblastoma patient sera that reacted with beta-tubulin I and III isoforms in neuroblastoma tissues did not react with beta-tubulin I and III isoforms found in normal brain tissue. Our findings indicate the occurrence of beta-tubulin peptides in neuroblastoma, which are immunogenic. The occurrence of immunogenic peptides in neuroblastoma may have utility in diagnosis and in immunotherapy of this aggressive childhood tumor. PMID- 11051244 TI - WT1 splicing alterations in Wilms' tumors. AB - Hereditary and sporadic forms of tumors are generally related to germ-line and somatic mutations of the same tumor suppressor gene. Unexpectedly, in Wilms' tumor, somatic mutations of the WT1 gene were found only occasionally in sporadic cases, although constitutional mutations of this gene are clearly associated with predisposition. It has been suggested that abnormal splicing may be another mode of somatic WT1 alteration. However, this idea was based on the analysis of a small series of tumors, precluding accurate evaluation of the frequency of such changes. To investigate WT1 changes at the somatic level in more detail, we analyzed the levels of the four isoform transcripts produced by alternative splicing events in a large series of 50 tumors, normal mature kidneys, and fetal kidneys. We characterized splicing alterations in 63% of sporadic Wilms' tumors. Moreover, taking into account the decreased and increased overall levels of WT1 mRNA, the percentage of sporadic tumors with changes in WT1 expression reached 90%. Whether and how these alterations of expression play a role in the tumorigenic process remain to be evaluated. PMID- 11051245 TI - Acquired expression of p27 is a favorable prognostic indicator in patients with hepatocellular carcinoma. AB - The p27 cyclin-dependent kinase inhibitor is a negative regulator of cell-cycle progression. In many human epithelial malignancies, decreased expression of p27 correlates with high grade, early recurrence, and poor prognosis. To evaluate the prognostic significance of p27 in hepatocellular carcinoma (HCC), we studied 54 HCCs along with corresponding nontumoral tissue. Immunohistochemistry (IHC) and Western blot (WB) analysis before and after immunoprecipitation with Cdk2 were performed on paraffin-embedded tissues and protein homogenates, respectively, to compare localization and expression of the p27 protein and to determine the total and active (Cdk2-bound) fractions of p27. Correlations were analyzed between IHC assessed levels of p27, survival, and major clinical and pathological variables. IHC revealed no p27 expression in the majority of hepatocytes from normal and cirrhotic liver, whereas 14 HCCs (26%) were high p27 expressers (>50% positive cells), 26 (48%) low expressers (<50% positive cells), and 14 (26%) negative. High IHC signals of p27 correlated with Cdk2-bound p27 as assessed by immunoprecipitation-WB; by contrast, WB alone displayed similar levels of p27 protein in all normal and tumoral samples. High IHC p27 expression correlated with prolonged survival (P = 0.027), whereas the presence of cirrhosis was associated with poor outcome (P = 0.029). We conclude that with respect to their nonneoplastic counterparts, a subset of HCCs acquire significant p27 expression and that high expression of p27 is a favorable independent prognostic parameter for HCC. PMID- 11051246 TI - Different patterns of allelic loss (loss of heterozygosity) in recurrent human pituitary tumors provide evidence for multiclonal origins. AB - Sporadic human pituitary tumors are benign adenomas of monoclonal origin. This implies that they arise from de novo somatic mutation(s) within a single pituitary cell. The availability of original and recurrent/regrown tumors from the same patient allowed testing of the prediction that recurrent/regrown tumors have identical genetic abnormalities as the original tumor sample. We used PCR amplification, from archival slide-extracted DNA, to allelotype microsatellite polymorphisms as an indication of clonality and confirmed this by X chromosome inactivation analysis in samples from women. Tumors from 33 of 49 (67%) patients with two or more specimens showed loss of heterozygosity (LOH) of at least one marker in at least one of their samples. Two patterns of LOH were observed. In pattern A in 14 of 33 (42%) of patients, the LOH pattern of the first tumor was preserved in the second recurrent sample, with some recurrent tumors also showing additional LOH. In these patients, the original and second tumors are presumed to arise from the same original clone with or without progressive accumulation of LOH. In pattern B [19 of 33 (58%) patients], LOH seen in the first tumor was not preserved in the second or subsequent tumors, as evidenced by retention of heterozygosity compared with the first tumor. The simplest explanation is that the second tumor, although still monoclonal, arises from another independently abnormal clone. This was confirmed by X chromosome inactivation analysis in all 11 women where this was informative. These results show that initial and recurrent tumors, of a benign tumor type, are frequently derived from separate independent clones. This suggests that either: (a) more than one abnormal clone is present from the outset though only one dominates; or (b) several clones arise independently at different times. In both scenarios, the initiating event(s) that predisposes to transformation might result in multiclonal hyperplasia, possibly as a consequence of exogenous stimulation. PMID- 11051247 TI - Nonhistological diagnosis of human cerebral tumors by 1H magnetic resonance spectroscopy and amino acid analysis. AB - We describe a multivariate analysis procedure to classify human cerebral tumors nonhistologically in vitro, combining the use of 1H magnetic resonance spectroscopy (MRS) with automatic amino acid analysis of biopsy extracts. Eighty one biopsies were obtained surgically and classified histologically in eight classes: high-grade astrocytomas (class 1, n = 19), low-grade astrocytomas (class 2, n = 10), normal brain (class 3, n = 9), medulloblastomas (class 4, n = 4), meningiomas (class 5, n = 18), metastases (class 6, n = 8), neurinomas (class 7, n = 9), and oligodendrogliomas (class 8, n = 4). Perchloric acid extracts were prepared from every biopsy and analyzed by high resolution 1H MRS and automatic amino acid analysis by ionic exchange chromatography. Intensities of 27 resonances and ratios of resonances were measured in the 1H MRS spectra, and 17 amino acid concentrations were determined in the chromatograms. Linear discriminant analysis provided the most adequate combination of these variables for binary classifications of a biopsy between any two possible classes and in multiple choice comparisons, involving the eight possible classes considered. Correct diagnosis was obtained when the class selected by the computer matched the histological diagnosis. In binary comparisons, consideration of the amino acid profile increased the percentage of correct classifications, being always higher than 75% and reaching 100% in many cases. In multilateral comparisons, scores were: high-grade astrocytomas, 80%; low-grade astrocytomas, 74%; normal brain, 100%; medulloblastomas, 100%; meningiomas, 94.5%; metastases, 86%; neurinomas, 100%; and oligodendrogliomas, 75%. These results indicate that statistical multivariate procedures, combining 1H MRS and amino acid analysis of tissue extracts, provide a valuable classifier for the nonhistological diagnosis of biopsies from brain tumors in vitro. PMID- 11051248 TI - High frequency of clonally related tumors in cases of multiple synchronous lung cancers as revealed by molecular diagnosis. AB - In patients with multiple synchronous lung tumors, discrimination of multicentric lung cancers from intrapulmonary metastasis is important for treatment decision, but this is sometimes difficult. The aim of this study was to retrospectively distinguish multicentric lung cancers from intrapulmonary metastases in 14 such cases by loss of heterozygosity (LOH) and p53 mutational status. DNA was extracted from microdissected tumor cells in paraffin-embedded archival tissue, and 3p14.2, 3p21, 3p25, 9p21, and 18q21.1 were investigated for LOH. Exons 5-8 of the p53 gene were examined for mutations by the PCR, followed by single-strand conformation polymorphism analysis and DNA sequencing. For cases with the same LOH pattern, we calculated a clonality index, the probability of the given LOH pattern when these tumors were hypothesized to be independent in origin. Eleven of 14 cases (79%) were thus diagnosed as having pulmonary metastasis and only one case as having genuinely multicentric lung cancers. Two cases presented difficulty in diagnosis. In several cases, the LOH patterns conflicted with p53 mutation patterns, suggesting that clonal evolution is directly affected by certain genetic changes. The combination of p53 with LOH helped increase both the sensitivity and specificity of the assay. PMID- 11051249 TI - Genomic aberrations in human hepatocellular carcinomas of differing etiologies. AB - We sought to assess whether genetic abnormalities in hepatocellular carcinoma differed in geographic locations associated with different risk factors. Comparative genomic hybridization (CGH) was applied to the genome-wide chromosomal analysis in 83 tumor samples from four different geographic origins. Samples were obtained from regions that differed in aflatoxin exposure: China (Hong Kong with low aflatoxin exposure and Shanghai with moderate aflatoxin exposure), Japan, and the United States (negligible aflatoxin exposure). Cases from Hong Kong and Shanghai were all hepatitis B virus (HBV) related, those from Japan were hepatitis C virus related, and those from the United States were HBV negative. In parallel, the mutational pattern of the whole p53 gene (exons 1-11) was also investigated in these cases. CGH revealed a complex pattern of chromosomal gains and losses, with the commonest aberration in each geographic location being chromosome 1q copy number gain (38-60%). Shanghai cases displayed the highest number of total aberrations per sample, with significant copy losses on 4q (75%), 8p (70%), and 16q (65%). Hepatitis C virus-related samples from Japan had a characteristically high incidence of 11q13 gain. p53 mutation(s) was detected in 23% of Hong Kong cases, 40% of Shanghai, 31% of Japan, but only 6% of the United States cases. The "aflatoxin-associated" codon 249 mutation was, however, identified only in samples from China (13% Hong Kong and 20% Shanghai). This finding, together with the highly aberrant pattern of genetic changes detected in the Shanghai series, is suggestive of the genotoxic effects of aflatoxin being more broadly based. It is also likely that there is a synergistic effect of HBV infection and high aflatoxin exposure in promoting hepatocellular carcinoma development. It appears from our CGH study that individual risk factors are indeed associated with distinct genetic aberrations, although changes in 1q gain appear common to all. PMID- 11051250 TI - Presence of human papilloma virus in tumor tissue from children with retinoblastoma: an alternative mechanism for tumor development. AB - Epidemiological studies have shown that the use of barrier methods of contraception is associated with a decreased incidence of papilloma virus infection and reduced risk of having a child with retinoblastoma. Thirty-nine primary retinoblastomas were analyzed for the presence of papilloma virus sequences. Tumor tissue sections were also used to assess the expression of the retinoblastoma protein and proliferative index. Papilloma sequences were detected in 14 of 39 (36%) tumors. Tumors in which viral sequences were detected were associated with a lower proliferative index (68% versus 78%; P = 0.015). Children with tumors containing viral sequences had a lower risk of extraocular disease (odds ratio, 9.0; 95% confidence interval, 1.6-49; P = 0.008) and a lower birth weight (2.9 versus 3.5 kg; P = 0.030). Based on these data, it is our hypothesis that papilloma viruses may play a role in the development of sporadic retinoblastoma. Detection of papilloma virus sequences and retinoblastoma protein in certain primary lesions suggests an alternative mechanism of tumor development for sporadic retinoblastoma. PMID- 11051251 TI - Racial differences in the prognostic usefulness of MUC1 and MUC2 in colorectal adenocarcinomas. AB - There is a need for new prognostic parameters that could add insights into the aggressiveness of tumors. Because the expression of two well-characterized mucin antigens, MUC1 and MUC2, in colorectal adenocarcinomas (CRCs) has been correlated with the aggressiveness of CRCs, we evaluated the prognostic value of the expression of MUC1 and MUC2 in CRCs collected from African-American and Caucasian patients. Expression of MUC1 and MUC2 was evaluated by immunohistochemistry in 166 archival CRC specimens collected from 58 African-American and 108 Caucasian patients that had been analyzed previously for nuclear accumulation of p53 (p53nac). Univariate Kaplan-Meier and multivariate Cox proportional hazards models were used to determine the prognostic significance of expression of MUC1 and MUC2 in these CRCs. MUC1 expression was more frequent in advanced stage CRCs, whereas MUC2 expression was higher in the mucinous type of CRCs. Although similar proportions of CRCs from African-Americans and Caucasians expressed MUC1 and MUC2, the MUC1 expression was found to be an indicator of high risk of death from CRC in Caucasians (hazard ratio, 2.03; P = 0.038) but not in African-Americans. Furthermore, Caucasians with CRCs exhibiting concomitant expression of MUC1 and p53nac demonstrated the lowest probability of overall survival (log rank test, P = 0.004). No prognostic value was found for MUC2 alone or in combination with p53nac in either group of patients. Expression of MUC1 in CRCs is a valuable indicator of poor prognosis in Caucasian patients. Additionally, combined evaluation of MUC1 and p53nac increases the ability to identify Caucasian patients with aggressive subtypes of CRC and may be useful in selecting or in developing novel therapeutic regimes. PMID- 11051252 TI - Loss of estrogen receptor (ER) expression in endometrial tumors is not associated with de novo methylation of the 5' end of the ER gene. AB - Normal endometrium, an estrogen-responsive tissue, expresses the estrogen receptor (ER) alpha gene. Loss of ER expression, the basis for which is currently unknown, is often seen in advanced stage, poor prognosis endometrial tumors. The ER gene undergoes de novo methylation with high frequency in a wide variety of human tumors, including ER-negative breast cancers. In this study, we used several bisulfite-based detection methods to assess whether loss of ER positivity in endometrial tumors is associated with aberrant methylation of the ER gene. Although extensive methylation of a 600-bp region at the 5' end of the gene was seen in two endometrial carcinoma cell lines, none of the 55 CpGs in this region was methylated in 25 of 26 ER-deficient endometrial carcinomas. PMID- 11051253 TI - The evolution of loss of heterozygosity on chromosome 17 during the progression to barrett's adenocarcinoma involves a unique combination of target sites in individual specimens. AB - We have previously identified thirteen common minimally deleted regions (MRs) on chromosome 17 in twelve Barrett's esophageal adenocarcinoma (BOA) specimens using 41 precisely mapped microsatellite markers (Dunn et al., Oncogene, 17: 987-993, 1999). The aim of the present study has been to identify the earliest sites of loss on this chromosome that arise and persist during the progression to BOA. This has been undertaken by the analysis of multiple carefully microdissected tissue samples from each of five esophagectomy specimens, several of which contained identifiable premalignant tissue. Our data demonstrate a stepwise accumulation of loss in each analyzed specimen, consistent with a single clonal pathway in four specimens and several coexisting pathways in one specimen. Several clonal anomalies (loss preceding heterozygosity and variable intrasample degrees of loss at different markers) were also observed. Within extensively deleted regions of the tumor (seen in three specimens), small deletions were detected in premalignant tissue, predominantly at the site of our identified MRs, and these losses were seen to expand and merge during the progression to BOA. Clonal losses at MRs were first detected in tissue showing early changes histologically, including Barrett's intestinal metaplasia and intermediate-grade dysplasia. Our results provide further support for many of the MRs we have previously identified, thereby adding to evidence for the existence of multiple novel cancer-associated genes on chromosome 17 involved in the development of BOA. PMID- 11051254 TI - Human breast cancer susceptibility to paclitaxel therapy is independent of Bcl-2 expression. AB - In laboratory studies, ectopic overexpression of the antiapoptotic protein Bcl-2 has been shown to result in resistance to the cytotoxic effects of many chemotherapeutic drugs. Furthermore, posttranslational modification of moderately expressed endogenous Bcl-2 has been correlated with susceptibility to paclitaxel treatment in vitro. To determine whether tumor expression of Bcl-2 protein correlates with response and ultimate outcome in vivo, we quantified Bcl-2 expression by immunohistochemical analysis of archived biopsy specimens from metastatic breast cancer patients treated with single-agent paclitaxel. The statistical association between the degree of Bcl-2 expression, objective tumor response, and clinical outcome was then determined. In patients (n = 39) whose tumors had low (< or = 10% cells positive) Bcl-2 levels by immunohistochemical analysis, the overall response (complete response + partial response) rate was 21% versus an overall response rate of 22% in patients (n = 36) with high (>10% cells positive) Bcl-2 expression (P = 0.92). In patients with low Bcl-2 expression, the median time to progression was 126 days [95% confidence interval (CI), 63-160 days]. This was not significantly different than the 105 days for patients with high tumor Bcl-2 expression (95% CI, 84-214 days). The median survival time from initiation of paclitaxel therapy for patients with low Bcl-2 expression was 663 days (95% CI, 456-1119 days) and was not significantly different than the 450 days (95% CI, 239-1058 days) observed for patients with high Bcl-2 expression. In conclusion, we found that in metastatic breast cancer, there is no significant association between tumor Bcl-2 expression and response to paclitaxel, median time to progression, or survival, suggesting that the main mechanism of paclitaxelinduced cytotoxicity in breast tumors is independent of Bcl-2 expression. PMID- 11051255 TI - Expression of prostate-specific membrane antigen in transitional cell carcinoma of the bladder: prognostic value? AB - The expression of Prostate-specific membrane antigen (PSMA) mRNA was assessed in the normal bladder urothelium (n = 9), transitional cell carcinoma (TCC) specimens (n = 52), TCC-derived cell lines (n = 3), and preoperative blood samples from TCC patients (n = 27). Specific PSMA mRNA was found in 100% of normal and malignant tissues and two cell lines. PSMA protein was detected in normal (n = 3) and malignant tissues (n = 4). Using a PSMA-specific substrate, PSMA enzymatic activity was found in two bladder cell lines and correlated with immunostaining. Seven of the 27 TCC preoperative blood samples were positive by reverse transcription-PCR. These preliminary results, obtained on a nonrandomized cohort of patients, correlated with tumor invasion (positive RT-PCR: 0% for pT < or = 2 versus 41% for pT > or = 3) and 2-year survival rate (81% in the PSMA negative group versus 29% in the PSMA-positive group). Although the clinical usefulness of this assay requires confirmation in larger prospective randomized trials, current preliminary results suggest that a blood-borne PSMA mRNA PCR assay may be a useful tool to predict a poor outcome in TCC patients. PMID- 11051256 TI - Prognostic significance of p53 alterations in patients with non-small cell lung cancer: a meta-analysis. AB - There is great controversy as to whether alteration of the p53 gene adversely affects survival of non-small cell lung cancer patients. The aim of this study was to qualitatively review the association between p53 alterations and patient outcome by reviewing published papers. Forty-three articles were used. Survival difference was combined by use of the DerSimonian-Laird method. p53 alteration was either detected as overexpression by the protein studies or as mutation by the DNA studies. The incidence of p53 alteration in DNA studies (381 of 1031; 37%) was lower than that in protein studies (1725 of 3579; 48%; P < 0.0001, chi2 test). The incidence of p53 overexpression and mutation in adenocarcinoma (36 and 34%) was significantly lower than that in squamous cell carcinoma (54 and 52%; P < 0.0001). Combined survival differences at 5 years (survival in patients with alteration minus that in patients without alteration) by protein and DNA studies were -9.1% (P = 0.0091) and -22.0% (P = 0.0026), respectively. The negative prognostic effect of p53 alteration was highly significant in patients with adenocarcinoma [-21.8% at 5 years (P = 0.0000039) by protein studies and -48.0% (P = 0.000031) by DNA studies] but not in patients with squamous cell carcinoma [ 15.6% (P = 0.4241) by protein studies and 2.0% (P = 0.8864) by DNA studies]. In the light of these results, p53 alteration was a significant marker of poor prognosis in patients with pulmonary adenocarcinoma. Whether p53 alteration also provides information that can alter treatment decisions should be asked in clinical trials. PMID- 11051257 TI - Cyclooxygenase-2 expression correlates with tumor neovascularization and prognosis in human colorectal carcinoma patients. AB - The role of cyclooxygenase-2 (COX-2) in tumor neovascularization of human colorectal carcinoma is yet to be delineated. One hundred colorectal carcinoma specimens were evaluated for COX-2 expression and CD34-stained microvessel density (MVD) by immunohistochemical methods. The relationships between COX-2 expression and clinicopathological feature of the patients, MVD, and survival time were analyzed. Increased COX-2 expression was significantly correlated with pathologically unfavorable findings such as tumor size (> 3.0 cm), tumor differentiation (poor, moderate > well differentiated), number of metastatic lymph nodes (24), and Dukes' stage (Dukes' B, C, and D). Larger number of microvessels congregated around the COX-2-expressing area, and the Spearman rank correlation test showed a strong correlation between COX-2 expression and tumor MVD (P < 0.0001). Patients with COX-2-positive tumors had a significantly (P = 0.037, by log-rank test) shorter survival time than those with negative tumors did. In the multivariate analysis, however, only Dukes' stage and number of metastatic lymph nodes remained as independent prognostic factors. Augmented tumor neovascularization may be one of the several effects of COX-2 responsible for poor prognosis in human colorectal carcinoma patients. PMID- 11051258 TI - Evaluation of serum KL-6, a mucin-like glycoprotein, as a tumor marker for breast cancer. AB - The utility of serum KL-6 as a tumor marker for breast cancer was evaluated in this study. The sera from 146 patients with breast cancer, 13 with benign breast disease, and 108 healthy individuals were measured for KL-6 titer using a sandwich enzyme immunoassay method. Carcinoembryonic antigen (CEA) and carbohydrate antigen 15-3 (CA15-3) titers were also tested in the same sera from the patients. The mean KL-6 titer of patients with primary breast cancer was 673 units/ml, which was significantly higher than that of benign and healthy individuals (P = 0.037 and P < 0.0001, respectively). The titer of patients with relapsed breast cancer was 1964 units/ml, which was also higher than that of primary cancer (P = 0.013). KL-6 titer was related to tumor stage, distant metastasis, and relapse site (P = 0.0053, P < 0.0001, and P = 0.0251, respectively). Using the cutoff value of 467 units/ml, the sensitivity of KL-6 was 31% for primary breast cancer (16% for stage I and 29% for stage II) and 73% for relapsed breast cancer (50% for local relapse and 89% for distant relapse). The specificity was 92%. The sensitivity of KL-6 was higher than that of CA15-3 and CEA. Combination of the three markers, followed by KL-6 and CEA, raised the sensitivity for primary breast cancer. Single use of KL-6 demonstrated a higher sensitivity than in each combination for relapsed breast cancer. In conclusion, serum KL-6 may be helpful for clinical use as a tumor marker for breast cancer, and it may play an important role, especially in the surveillance of disease relapse. PMID- 11051259 TI - Prognostic significance of p27KIP1 protein and ki-67 growth fraction in non-small cell lung cancers. AB - We immunohistochemically examined specimens of 215 surgically resected non-small cell lung cancers (NSCLCs) for p27KIP1 protein (p27) expression and the growth fraction determined by the Ki-67 labeling index (LI). The NSCLCs analyzed showed considerable heterogeneity in both p27 and Ki-67 LIs; 25 of 207 (13%) lacked p27 expression (p27 LI < 5%), and 116 of 215 (54%) showed a high Ki-67 LI (>30%). The p27 LI was not significantly associated with the Ki-67 LI. A chi2 test showed that loss of p27 expression was inversely correlated with smoking (P = 0.01) and that a high Ki-67 LI was significantly associated with male gender, squamous cell carcinoma histology, and smoking (P < 0.0001 each). Prognostic values of p27 and Ki-67 expression were evaluated in 109 tumors of postsurgical pathological stages I and II. Patients with tumors lacking p27 expression survived for a significantly shorter time than patients with tumors expressing p27 (5-year survival rates, 38% and 68%, respectively; P = 0.02). Patients with tumors having a high Ki-67 LI survived for a significantly shorter time than patients with tumors having a low Ki-67 LI (5-year survival rates, 48% and 78%, respectively; P = 0.005). Multivariate analysis showed that loss of p27 expression tended to be an unfavorable prognostic factor (P = 0.054), whereas a high Ki-67 LI was a significant and independent unfavorable prognostic factor (P = 0.004). When analyzed by cell types, loss of p27 expression was a significant and independent unfavorable prognostic factor in squamous cell carcinomas (P = 0.01), whereas a high Ki-67 LI was a significant and independent unfavorable prognostic factor in nonsquamous cell carcinomas (P = 0.007). We further evaluated the importance of p27 expression in clinical outcome in combination with the Ki-67 LI and ras p21 protein (ras) expression, which we previously reported as an important prognostic factor in NSCLCs. Patients with tumors lacking p27 expression and having a high Ki-67 LI survived for a significantly shorter time than those with tumors expressing p27 and having a high Ki-67 LI (5-year survival rates, 17% and 52%, respectively; P = 0.003). Patients with p27-negative and ras-positive tumors survived for a significantly shorter time than those with both p27- and ras positive tumors (5-year survival rates, 0% and 38%, respectively; P < 0.0001). These results indicate the pivotal roles of p27 and Ki-67 expression in the clinical outcome of NSCLCs. PMID- 11051260 TI - Study of dose escalation and sequence switching of administration of the combination of docetaxel and doxorubicin in advanced breast cancer. AB - The objectives of the present study were to evaluate whether a schedule-dependent pharmacokinetic and/or pharmacodynamic interaction exists between two sequences of docetaxel and doxorubicin administration and to determine the maximal tolerated dose (MTD) of this combination. Patients with chemotherapy-naive metastatic or recurrent advanced breast cancer were enrolled. In the crossover design, tandem dose escalation of docetaxel and doxorubicin was performed. Docetaxel, in doses ranging from 50-70 mg/m2, was administered for 1 h by drip infusion either just before or after a 5-min bolus i.v. injection of doxorubicin at dosages from 40-50 mg/ m2. The sequence of drug administration was switched after the first course in each patient, and the sequence of drug administration thereafter depended on the patient's choice. Twenty-five patients were initially assessable for toxicity. The MTD in the sequence of doxorubicin after docetaxel was 40 and 50 mg/m2, respectively, with the dose-limiting toxicity of neutropenia. On the other hand, the MTD of the sequence of docetaxel after doxorubicin was 70 and 50 mg/m2, respectively. The dose-limiting toxicities in this sequence were neutropenia and diarrhea. Duration of grade 4 neutropenia in the sequence of docetaxel followed by doxorubicin was significantly longer than that in the alternate sequence (P = 0.0062). However, there was no difference in pharmacokinetic parameters of docetaxel, doxorubicin, and doxorubicinol between the two sequences. The sequence of 50 mg/m2 doxorubicin followed by 60 mg/m2 docetaxel is recommended for subsequent clinical trials for practical reasons. PMID- 11051261 TI - Analysis of genetic polymorphism in NQO1, GST-M1, GST-T1, and CYP3A4 in 469 Japanese patients with therapy-related leukemia/ myelodysplastic syndrome and de novo acute myeloid leukemia. AB - Several genetic polymorphisms in metabolic activation or detoxification enzymes have been associated with susceptibility to therapy-related leukemia and myelodysplastic leukemia (TRLIMDS). We analyzed gene polymorphisms of NAD(P)H:quinone oxidoreductase (NQOl), glutathione S-tranferase (GST)-MI and -TI, and CYP3A4, the enzymes of which are capable of metabolizing anticancer drugs, in 58 patients with TRL/MDS and in 411 patients with de novo acute myeloid leukemia (AML). Homozygous Ser/Ser genotype of NQOl at codon 187, causing loss of function, was more frequent in the patients with TRLIMDS (14 of 58, 24.1%; OR = 2.62) than in those with de novo AML (64 of 411, 15.6%), and control (16 of 150, 10.6%; P = 0.002). Allelic frequencies of NQOJ were different between TRL/ MDS and de novo AML (P = 0.01). In GST-MJ and -Ti, the incidence of homologous deletion was similar among the three groups. The polymorphism of the 5' promoter region of CYP3A4 was not found in persons of Japanese ethnicity. These results suggest that the NQOJ polymorphism is significantly associated with the genetic risk of TRLIMDS. PMID- 11051262 TI - Overexpression of retinoblastoma protein predicts decreased survival and correlates with loss of p16INK4 protein in gallbladder carcinomas. AB - This study was designed to determine whether the level of retinoblastoma protein (pRb) expression predicts tumor progression and prognosis in gallbladder carcinomas (GBCs) and the relationship between pRb and pl6INK4 protein expression. The expression of these two proteins was evaluated immunohistochemically in 37 tumors from 36 patients with GBC. pRb loss and overexpression were observed in 5 (13.5%) and 18 (48.6%) of the 37 tumors, respectively. Both pRb loss and overexpression were significantly correlated with advanced TNM stage, lymph node metastasis, and tumor perineural invasion. Moreover, pRb overexpression was significantly associated with decreased overall survival (P = 0.001; log-rank test). Further analysis indicated that the influence of pRb overexpression on survival was independent of TNM stage and lymph node metastasis. Loss of p16INK4 protein was observed in 28 of the 37 GBCs (75.7%), but was not significantly associated with any clinicopathological factors or survival. pRb overexpression was significantly associated with the loss of p161NK4 protein (P < 0.0001). These results suggest that pRb overexpression significantly predicts decreased survival in GBCs. PMID- 11051263 TI - Prostate cancer gene therapy: comparison of adenovirus-mediated expression of interleukin 12 with interleukin 12 plus B7-1 for in situ gene therapy and gene modified, cell-based vaccines. AB - We have documented previously that adenovirus-mediated interleukin 12 (IL-12) gene therapy is effective for orthotopic tumor control and suppression of pre established metastases in a preclinical prostate cancer model (Y. Nasu et al., Gene Ther., 6: 338-349, 1999). In this report, we directly compare the effectiveness of an adenovirus that expresses both IL-12 and the costimulatory molecule B7-1 (AdmIL12/B7) with one that expresses IL-12 alone (AdmIL-12) using the poorly immunogenic RM-9 orthotopic murine model of prostate cancer. We document AdmIL-12/B7-mediated secretion of IL-12 and increased surface expression of B7-1 in infected RM-9 tumor cells. A significant reduction in orthotopic tumor size and increased survival was demonstrated in mice treated with a single orthotopic injection of AdmIL-12/B7 compared with AdmIL-12 or controls. Six of 19 animals treated with AdmIL-12/B7 survived long term with apparent eradication of the primary tumor in contrast to one of 38 animals in the AdmIL-12-treated group. Orthotopic treatment of tumors with both vectors led to an infiltration of both CD4+ and CD8+ immunoreactive cells, with AdmIL-12/B7 treatment having a more prolonged infiltration of CD8+ cells. AdmIL-12/B7 was also more effective than AdmIL-12 or controls at suppression of pre-established metastases. We further developed a vaccine model based on s.c. injection of infected, irradiated RM-9 cells and found that both AdmIL-12 and AdmIL-12/B7 are effective at suppressing the development and growth of challenge orthotopic tumors using this protocol. PMID- 11051265 TI - Pediatric rhabdomyosarcoma cell lines are resistant to Fas-induced apoptosis and highly sensitive to TRAIL-induced apoptosis. AB - Seven pediatric rhabdomyosarcoma (RMS) cell lines were resistant to the induction of apoptosis via the Fas death receptor. In contrast, four of seven lines (RD, Rh1, Rh18, and Rh30) were highly sensitive to tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL). TRAIL induced apoptosis within 4 h and also reduced clonogenic survival, both reversible by caspase inhibitors. DR5 (but not DR4) was expressed at high level in all cell lines. Expression of the decoy receptors DcR1 and DcR2 did not correlate with TRAIL sensitivity. All RMS lines expressed the adapter molecule FADD, and six of seven expressed procaspase-8. Expression of the inhibitory proteins c-FLIPL and c-FLIPs was high in three TRAIL sensitive (RD, Rh1, and Rh30) and two TRAIL-resistant (Rh28 and Rh41) lines. All RMS lines expressed Bid and procaspases-3, -6, -7, and -9. Procaspases-8 and -10 were highest in TRAIL-sensitive RMS (RD, Rh1, and Rh30), and procaspase-10 was not expressed in Rh18, Rh36, or Rh41. TRAIL induced loss of mitochondrial membrane potential in TRAIL-sensitive Rh1 but not in TRAIL-resistant Rh41 cells. There was no correlation between expression of members of the Bcl-2 family (Bcl 2, Bcl-xL, Bax, and Bak) and TRAIL sensitivity. TRAIL-sensitive Rh18 expressed procaspase-8 in the absence of procaspase-10 and c-FLIP, and procaspase-10 was not detected in TRAIL-resistant Rh41 in the presence of procaspase-8 and c-FLIP. Data suggest that caspase-8 may be sufficient to deliver the TRAIL-induced apoptotic signal in the absence of both caspase-10 and c-FLIP (Rh18) but not in the presence of c-FLIP (Rh41). In RD, Rh1, and Rh30, the presence of c-FLIP may require amplification of the apoptotic signal via caspase-10. PMID- 11051264 TI - Antitumor activity of temozolomide combined with irinotecan is partly independent of O6-methylguanine-DNA methyltransferase and mismatch repair phenotypes in xenograft models. AB - The activity of temozolomide combined with irinotecan (CPT-11) was evaluated against eight independent xenografts (four neuroblastomas, three rhabdomyosarcomas, and one glioblastoma). In all studies, temozolomide was administered p.o. daily for 5 consecutive days/cycle, found in preliminary studies to be the optimal schedule for administration. Irinotecan was administered i.v. for 5 days for 2 consecutive weeks/cycle. Treatment cycles were repeated every 21 days for a total of three cycles over 8 weeks. In combination, temozolomide and CPT-11 induced complete responses in four neuroblastomas, two rhabdomyosarcomas, and the glioblastoma line. The activity of the combination was significantly greater than the activity of either agent administered alone in four tumor lines. Of interest, the interaction appeared independent of tumor MGMT or mismatch repair phenotype, suggesting that the mechanism of synergy may be independent of O6-methylation by temozolomide. Pharmacokinetic studies indicated no detectable interaction between these two agents. Further, coadministration of CPT-11 appeared to reduce the toxicity of temozolomide in tumor-bearing mice. PMID- 11051266 TI - The efficacy of a broad-spectrum sunscreen to protect engineered human skin from tissue and DNA damage induced by solar ultraviolet exposure. AB - Sunscreens are known to protect against sunlight-induced erythema and sunburn, but their efficiency at protecting against skin cancer is still a matter of debate. Specifically, the capacity of sunscreens to prevent or reduce tissue and DNA damage has not been thoroughly investigated. The present study was undertaken to assess the ability of a chemical broad-spectrum sunscreen to protect human skin against tissue and DNA damage after solar UV radiation. Engineered human skin was generated and either treated or not with a broad-spectrum SPF 30 sunscreen and exposed to increasing doses of simulated sunlight (SSL). Immediately after irradiation, histological, immunohistochemical, and molecular quantitative analyses were performed. The unprotected irradiated engineered human skin showed significant epidermal disorganization accompanied by a complete absence of laminin deposition. The sunscreen prevented SSL-induced epidermal damage at low doses and allowed laminin deposition at almost all SSL doses tested. The frequencies of cyclobutane pyrimidine dimers, pyrimidine (6-4) pyrimidone photoproducts, and photooxidative lesions measured by alkaline gel electrophoresis and radioimmunoassay were significantly reduced by the sunscreen. Thus, tissue and DNA damage may provide excellent quantitative end points for assessing the photoprotective efficacy of sunscreens. PMID- 11051267 TI - Cyclic GMP mediates apoptosis induced by sulindac derivatives via activation of c Jun NH2-terminal kinase 1. AB - Sulindac sulfone (Exisulind) induces apoptosis and exhibits cancer chemopreventive activity, but in contrast to sulindac, it does not inhibit cyclooxygenases 1 or 2. We found that sulindac sulfone and two potent derivatives, CP248 and CP461, inhibited the cyclic GMP (cGMP) phosphodiesterases (PDE) 2 and 5 in human colon cells, and these compounds caused rapid and sustained activation of the c-Jun NH2-terminal kinase 1 (JNK1). Rapid activation of stress-activated protein/ERK kinase 1 (SEK1) and mitogen-activated protein kinase kinase kinase (MEKK1), which are upstream of JNK1, was also observed. Other compounds that increase cellular levels of cGMP also activated JNK1, and an inhibitor of protein kinase G (PKG), Rp-8-pCPT-cGMPS, inhibited JNK1 activation by the sulindac sulfone derivatives. Expression of a dominant-negative JNK1 protein inhibited CP248-induced cleavage of poly(ADP-ribose) polymerase, a marker of apoptosis. Thus, it appears that sulindac sulfone and related compounds induce apoptosis, at least in part, through activation of PKG, which then activates the MEKK1-SEK1-JNK1 cascade. These studies also indicate a role for cGMP and PKG in the JNK pathway. PMID- 11051268 TI - Neutron radiation enhances cisplatin cytotoxicity independently of apoptosis in human head and neck carcinoma cells. AB - Recent advances in combined modality treatment of locally advanced head and neck cancer have improved local and regional disease control and survival with better functional outcome. However, the local and regional failure rate after radiation therapy is still high for tumors that respond poorly to cisplatin-based neoadjuvant chemotherapy. This clinical observation suggests a common biological mechanism for resistance to cisplatin and photon irradiation. In this report, we investigated the molecular basis underlying cisplatin resistance in head and neck squamous carcinoma (HNSCC) cells and asked if fast neutron radiation enhances cisplatin cytotoxicity in cisplatin-resistant cells. We found that cisplatin sensitivity correlates with caspase induction, a cysteine proteinase family known to initiate the apoptotic cell death pathway, suggesting that apoptosis may be a critical determinant for cisplatin cytotoxicity. Neutron radiation effectively enhanced cisplatin cytotoxicity in HNSCCs including cisplatin-resistant cells, whereas photon radiation had little effect on cisplatin cytotoxicity. Interestingly, neutron-enhanced cisplatin cytotoxicity was associated neither with apoptosis nor with cell cycle regulation, as determined by caspase activity assay, annexin V staining, and flow cytometric analysis. Taken together, the present study provides a molecular insight into cisplatin resistance and may also provide a basis for more effective multimodality protocols involving neutron radiation for patients with locally advanced head and neck cancer. PMID- 11051269 TI - Temozolomide delivered by intracerebral microinfusion is safe and efficacious against malignant gliomas in rats. AB - Intracerebral microinfusion (ICM) is an innovative technique of delivering therapeutic agents throughout large portions of the brain that circumvents the blood-brain barrier, minimizes systemic toxicity, and provides a homogeneous distribution of the infused agent. Temozolomide is a novel methylating agent with proven efficacy against malignant gliomas (MGs) after systemic administration but with dose-limiting myelotoxicity. Because MGs rarely metastasize, systemic drug delivery is unnecessary. Therefore, we evaluated the efficacy and toxicity of ICM with temozolomide in an athymic rat model of human MGs. Treatment of rats by ICM with temozolomide 3 days after intracerebral challenge with D54 human MG xenograft increased median survival by 128% compared with rats treated by ICM with saline, by 113% compared with rats treated with i.p. saline, and by 100% compared with rats treated with i.p. temozolomide (P < 0.001). Delay of treatment until 9 days after tumor challenge still resulted in a 23% increase in median survival in rats treated by ICM of temozolomide compared with rats treated with i.p. temozolomide. In addition, overall, 21.7% of rats treated by ICM with temozolomide survived for > 100 days without clinical or histological evidence of tumor. The dose of temozolomide delivered by ICM in this study was limited only by drug solubility, and no neurological or systemic toxicity could be attributed to ICM with temozolomide. Therefore, ICM of temozolomide may offer significant advantages in the treatment of MGs. PMID- 11051270 TI - Schedule-dependent activity of temozolomide plus CPT-11 against a human central nervous system tumor-derived xenograft. AB - Temozolomide, an imidazole tetrazinone, and CPT-11, a camptothecin derivative, have previously been shown to have anti-central nervous system tumor activity in laboratory and clinical studies. The current experiments were designed to evaluate the activity of temozolomide plus CPT-11 against a malignant glioma derived xenograft, D-54 MG, growing s.c. in athymic nude mice. The initial schedule of i.p. drug administration was temozolomide at 0.1 LD10 on day 1 and CPT-11 at 0.1 LD10 on days 1-5 and 8-14. The combination of these two agents produced greater than additive activity against D-54 MG. This enhanced activity was maintained when the initial administration of CPT-11 was delayed to day 3 or day 5. However, when CPT-11 was administered first on day 1 using 0.5 LD10 (for the single dose schedule) followed by temozolomide (0.1 LD10) 5 h, 3 days, or 5 days later, the enhancement of activity was substantially reduced. These results demonstrate that the combination of temozolomide plus CPT-11 displays a schedule dependent enhancement of antitumor activity, suggest a mechanistic explanation for the enhanced activity, and provide the rationale for a Phase I trial of this regimen. PMID- 11051271 TI - Effective treatment of metastatic MDA-MB-435 human estrogen-independent breast carcinomas with a targeted cytotoxic analogue of luteinizing hormone-releasing hormone AN-207. AB - A highly potent derivative of doxorubicin, 2-pyrrolinodoxorubicin (AN-201), was linked to [D-Lys6]luteinizing hormone-releasing hormone (LH-RH) to form a cytotoxic analogue, AN-207, that can be targeted to LH-RH receptors. The effects of AN-207 were investigated in MDA-MB-435 human estrogen-independent breast carcinomas, which express LH-RH receptors. In experiment 1, nude mice bearing orthotopically implanted tumors received a single i.v. injection of AN-207, AN 201, or the carrier at 250 nmol/kg. Five weeks after administration of AN-207, tumor volume was significantly decreased by 66% (P < 0.001) and tumor burden by 71% (P < 0.05) as compared with controls, but no significant effects occurred in other groups. Six of eight (75%) control animals and 37.5% of mice treated with AN-201 developed metastases in the lymph nodes, whereas no lymphatic spread was found in any of the mice that received injections of AN-207. The antitumor effect of AN-207 could be partially blocked by pretreatment of the tumor-bearing mice with high doses of agonist [D-Trp6]LH-RH, which suggests that AN-207 acts on LH RH receptors on tumors. The mortality due to toxicity was 25% in the group receiving AN-201 and 12.5% in the AN-207-treated group. Radioligand binding assays revealed the presence of high-affinity binding sites for LH-RH on tumor membranes, and mRNA for LH-RH receptors was demonstrated by reverse transcription PCR. In experiment 2, two i.v. injections of AN-207 or AN-201 at 150 nmol/kg were given on days 0 and 28 to mice bearing orthotopic xenografts of MDA-MB-435. The outcome of the treatment was similar to that observed in experiment 1, but without any toxicity-related deaths. Tumor growth inhibition and prevention of metastatic disease suggest that cytotoxic LH-RH analogue AN-207 could be considered for the treatment of human estrogen-independent breast cancers expressing receptors for LH-RH. PMID- 11051272 TI - Correspondence re: M. Fanciulli et al., Energy metabolism of human LoVo colon carcinoma cells: correlation to drug resistance and influence fo lonidamine. Clin. Cancer Res., 6: 1590-1597, 2000. PMID- 11051273 TI - Correspondence re: G. C. de Gast et al., Phase I trial of combined immunotherapy with subcutaneous granulocyte macrophage colony-stimulating factor, low-dose interleukin 2, and interferon alpha in progressive metastatic melanoma and renal cell carcinoma. Clin. Cancer Res., 6: 1267-1272, 2000. PMID- 11051274 TI - Gene therapy in experimental autoimmune encephalomyelitis. AB - Gene therapy traditionally has been associated with "gene replacement." where exogenous recombinant DNA is introduced ex vivo into somatic cells that are then introduced back into the patient as a way to correct an inherited genetic defect. However, several novel gene therapy strategies for treating autoimmune diseases recently have emerged. Strategies involving the use of several types of DNA vaccines, the application of various viral vectors, and the use of diverse cellular vectors have shown promise in inhibiting autoimmune-mediated inflammation and repairing tissue damaged as a result of autoimmune attack. In the current review, we examine and discuss the development and proposed use of emerging gene therapy strategies for the treatment of autoimmune disease with specific emphasis on experimental autoimmune encephalomyelitis (EAE), an animal model widely used in multiple sclerosis (MS) research. PMID- 11051275 TI - Gene therapy for autoimmune disorders. AB - Although many autoimmune disorders do not have a strong genetic basis, their treatment may nevertheless be improved by gene therapies. Most strategies seek to transfer genes encoding immunomodulatory products that will alter host immune responses in a beneficial manner. Used in this fashion, genes serve as biological delivery vehicles for the products they encode. By this means gene therapy overcomes obstacles to the targeted delivery of proteins and RNA, and improves their efficacy while providing a longer duration of effect, and, potentially, greater safety. Additional genetic strategies include DNA vaccination and the ablation of selected tissues and cell populations. There is considerable evidence from animal studies that gene therapies work: examples include the treatment of experimental models of rheumatoid arthritis, multiple sclerosis, diabetes, and lupus. Pre-clinical success in treating animal models of rheumatoid arthritis has led to the first clinical trial of gene therapy for an autoimmune disease. In this Phase I study, a cDNA encoding the interleukin-1 receptor antagonist was transferred to the knuckle joints of patients with advanced rheumatoid arthritis. Two additional clinical trials are in progress. It is likely that gene therapy will provide effective new treatments for a wide range of autoimmune disorders. PMID- 11051276 TI - Expression of perforin and Fas ligand mRNA in the liver of viral hepatitis. AB - Cytotoxic T lymphocytes (CTLs) play an important role in the pathogenesis of viral hepatitis. We studied the expression of mRNAs of perforin and Fas ligand (Fas-L) in biopsy specimens from chronic hepatitis B (CHB) (15 cases) and hepatitis C (CHC) patients (13 cases). Both perforin and Fas-L mRNAs were detected in all cases of both CHB and CHC. No messages were detected in the control livers from two cases of fatty liver, a case of Gilbert's syndrome, and a case of Dubin-Johnson syndrome. Semiquantitative analysis revealed a positive correlation between the intensity of perforin and Fas-L mRNAs in both CHB and CHC. In CHB, the intensity of both perforin and Fas-L mRNAs showed a positive correlation with the histological activity and serum alanine aminotransferase level, while the correlation was not apparent in CHC. These results suggest that both perforin and Fas/Fas-L systems are involved in the pathogenesis of liver cell injury of CHB and CHC. PMID- 11051277 TI - Calorie restriction decreases proinflammatory cytokines and polymeric Ig receptor expression in the submandibular glands of autoimmune prone (NZB x NZW)F1 mice. AB - Calorie restriction or fish oil (enriched in n-3 fatty acids) supplementation ameliorates glomerulonephritis and Sjogren's syndrome lesions in (NZB x NZW)F1(B/W) mice. Enhanced proinflammatory cytokine expression and deposition of immune complexes are the important pathological events in the development of Sjogren's syndrome. In the present study, we have examined the effect of calorie restriction and fish oil supplementation on the expression of key inflammatory cytokines [gamma interferon (INF-gamma), interleukin-10 (IL-10), and IL-12] and polymeric immunoglobulin (Ig) receptor (pIgR) (receptor for IgA and IgM) and the secretion of Ig in the submandibular glands (SMG) of B/W mice. Weanling B/W mice were fed either ad libitum (AL) or calorie restricted (CR) (40% less calories than AL) diet supplemented with 5% corn oil (CO) or 5% fish oil (FO) until 4 or 9 months of age. The SMGs were removed and a portion of the tissue used for semiquantitive determinations of IFN-gamma, IL-10, IL-12 (p40), and pIgR mRNA. The remaining SMG tissue was fragmented and cultured for 7 days and the culture supernatants assayed for IgA, IgM, and IgG2a levels by enzyme-linked immunosorbent assay. Results revealed a significant increase in the expression of IFN-gamma, IL-10, and IL-12 mRNA with age in AL fed mice, whereas CR fed mice maintained their levels to near those seen in young animals regardless of the dietary fat. PIgR mRNA expression also remained unaltered in CR animals irrespective of age and dietary fat, while it was found significantly increased in AL fed mice. CR significantly inhibited the elevated levels of IgA and IgG2a seen in aged mice. Interestingly, CR also influenced the Ig level in young animals. In summary, these results indicate that amelioration of autoimmune disease by CR in B/W mice is possibly mediated by the lowered mRNA expression of IFN-gamma, IL-10, IL-12, and pIgR and the reduced Ig secretion. PMID- 11051279 TI - Immune aberrations in B and T lymphocytes derived from chronic urticaria patients. AB - To investigate the pathophysiology of chronic urticaria (CU) in light of the abundant evidences that it is an autoimmune disease and to define some cellular markers in B/T lymphocytes that could be of pathogenic significance, we investigated 14 patients suffering from CU, compared to 7 contact dermatitis patients and 10 normal control individuals. We tested the expression of CD5, B7.1 (CD80), CD23, and CD25 on B cells and of CD(40L)) and CD25 on T cells from all studied individuals. We also tested B cell proliferation upon their activation followed by dexamethazone induced inhibition of proliferation. The expression of bcl-2 protein in activated lymphocytes from normal individuals was compared to that of contact dermatitis and CU patients. CD(40L) expression was found significantly higher on in (vitro activated CD3 [with phorbol myristate acetate (PMA) and ionomycine Ca2+ at 12 hr of culture] from CU patients compared to that of contact dermatitis and normal individuals. Whereas the proliferation of activated B cells from CU patients was higher, dexamethazone-induced inhibition of B cell proliferation was found statistically similar in both groups yet lower in B cells from most severe CU patients. We demonstrate a higher bcl-2 expression in activated B and T lymphocytes of severe CU patients compared to that of moderate CU and both contact dermatitis and normal individuals. The increased expression of CD(40L) on activated T cells might play a role in the polyclonal increased B cell proliferation of CU patients. This increased proliferation accompanies the finding that activated B and T lymphocytes from these patients demonstrate increased bcl-2 expression. The resistance of some B cells to dexamethazone-induced inhibition of proliferation encourages us to investigate the possibility that B cells from some CU patients might develop rescue mechanisms from activated cell death. These findings provide further evidence that CU is indeed an autoimmune disease, although its precise nature has yet to be fully elucidated. PMID- 11051278 TI - IL-10 secretion and sensitivity in normal human intestine and inflammatory bowel disease. AB - Interleukin-10 (IL-10) deficiency in gene knockout mice causes chronic enterocolitis. We hypothesized that inflammation in human inflammatory bowel disease might result from innate alterations in the IL-10 pathway. Serum, supernatants, and mRNA of peripheral blood mononuclear cells (PBMC) and lamina propria mononuclear cells (LPMC) derived from inflamed (LPMC-i) and noninflamed colonic mucosa (LPMC-ni) were collected from patients with Crohn's colitis, ulcerative colitis, and controls. IL-10 protein concentrations and IL-10 mRNA were examined in response to PMA/CD3 or PHA stimulation. The response to rhIL-10 was assessed by inhibition of tumor necrosis factor-alpha (TNF-alpha), IL-6, and interferon-gamma (IFN-gamma) production. Serum IL-10 levels of inflammatory bowel disease (IBD) patients were within the normal range. IL-10 concentrations in supernatants from LPMC-i were significantly lower than from LPMC-ni or PBMC. No difference was seen between samples from ulcerative colitis and Crohn's disease. IL-10 mRNA was detected in 0/4 LPMC-i samples compared to 1/6 LPMC-ni and 6/6 PBMC. RhIL-10 inhibited TNF-alpha, IL-6, and IFN-gamma synthesis in PBMC. This effect was strongly diminished in LPMC. Disease-specific alterations were not detected. Our data suggest that LPMC derived from inflamed colonic mucosa have a reduced ability to produce and to respond to rhIL-10. A disease-specific alteration in the IL-10 pathway, however, was not found. PMID- 11051280 TI - T-cell activation and differentiation are regulated by TNF during murine DBA/2- >B6D2F1 intestinal graft-versus-host disease. AB - Administration of a tumor necrosis factor (TNF) inhibitor-encoding adenoviral vector decreases the severity of colonic inflammation in a DBA/2-->B6D2F1 murine model of colonic graft-versus-host disease (GVHD). The present studies evaluated the effect of TNF blockade on the splenic and colonic T-cell responses. cDNA encoding an artificial fusion protein consisting of the extracellular domain of the human 55-kDa receptor for TNF fused to a mouse IgG heavy chain was subcloned into an E1a-deficient adenoviral vector. Following transfer of DBA/2 T cells and bone marrow cells into irradiated B6D2F1 mice, the mice then received either the control adenovirus or the TNF inhibitor-encoding adenovirus. Splenic and colonic lymphocytes were isolated, stained with anti-H-2b, anti-H-2d, anti-CD3, anti-CD4, anti-CD8, and anti-CD45RB antibodies, and analyzed by flow cytometry. Splenic and colonic lymphocyte cytokine profiles also were assessed. More colonic T cells of donor origin were isolated from the control adenovirus recipients than from recipients of the TNF inhibitor encoding adenovirus (P = .027). Fewer CD4+ and CD8+ T cells were observed in colon but not in the spleen in the TNF inhibitor recipients. Fewer CD45RBlow (memory) T cells were observed in the CD4+ colonic lymphocytes isolated from the TNF inhibitor recipients than from controls. Importantly, lower levels of interleukin-2(IL-2) and interferon-gamma (INF-gamma) but not of IL-4 were observed in the lamina propria lymphocyte RNA isolated from the TNF inhibitor recipients. Infiltration and expansion of donor T cells and T cell activation in the colon appear to be regulated by TNF during murine DBA/2 - > B6D2F1 gut GVHD. PMID- 11051281 TI - Preservation of sight in diabetes: developing a national risk reduction programme. AB - BACKGROUND: Early treatment for diabetic retinopathy is effective at saving sight, but dependent on pre-symptomatic detection. Although 60% of people with diabetes have their eyes examined annually, few UK health authorities have systematic programmes that meet the British Diabetic Association's standards for sensitivity (> 80%) and specificity (> 95%). Screening is generally performed by general practitioners and optometrists, with some camera-based schemes, operated by dedicated staff. The National Screening Committee commissioned a group to develop a model and cost estimates for a comprehensive national risk-reduction programme. OPHTHALMOSCOPY: Evidence indicates that direct ophthalmoscopy using a hand-held ophthalmoscope does not give adequate specificity and sensitivity, and should be abandoned as a systematic screening technique. Indirect ophthalmoscopy using a slit lamp is sensitive and specific enough to be viable, and widespread availability in high street optometrists is an advantage, but the method requires considerable skill. PHOTOGRAPHIC SCHEMES: The principal advantage of camera-based screening is the capturing of an image, for patient education, review of disease progression, and quality assurance. Digital cameras are becoming cheaper, and are now the preferred option. The image is satisfactory for screening and may be transmitted electronically. With appropriate training and equipment, different professional groups might participate in programme delivery, based on local decisions. COST ISSUES: Considerable resources are already invested in ad hoc screening, with inevitable high referral rates incurring heavy outpatient costs. Treatment for advanced disease is expensive, but less likely to be effective. The costs of a new systematic screening and treatment programme appear similar to current expenditure, as a result of savings in treatment of late-presenting advanced retinopathy. CONCLUSION: A systematic national programme based on digital photography is proposed. PMID- 11051282 TI - Evaluation of a holistic treatment and teaching programme for patients with Type 1 diabetes who failed to achieve their therapeutic goals under intensified insulin therapy. AB - AIMS: To evaluate a treatment and teaching programme including psychosocial modules for patients with Type 1 diabetes mellitus on intensified insulin therapy who failed to achieve their treatment goals despite participation in standard programmes. METHODS: The 5-day inpatient programme comprises small groups of 4-6 patients, focusing on individual needs and problems. Beyond the teaching lessons (most topics are deliberately chosen by the patients), the programme provides intensive group discussions and offers individual counselling concerning motivational aspects, psychosocial problems and coping strategies. Of the first consecutive 83 participants, 76 were re-examined after 17.5 +/- 5.5 months (range 9-31 months). RESULTS: At follow-up, HbA1c was not improved compared to baseline (8.0 +/- 1.3% vs. 8.1 +/- 1.5%). However, the incidence of severe hypoglycaemia per patient/year (glucose i.v., glucagon injection) was substantially decreased: 0.62 +/- 1.5 episodes at baseline compared to 0.16 +/- 0.9 at follow-up (P < 0.001). Twenty-six per cent of the patients at baseline, and 4% at re-examination had experienced at least one episode of severe hypoglycaemia during the preceding year (P < 0.001). Sick leave days per patient/year decreased from 17.0 +/- 38.5 7.7 +/- 13.6 days (P < 0.05). Patients improved their perceptions of self efficacy, their relationship to doctors and felt less externally controlled (P < 0.001). The majority of patients perceived an improved competence regarding diet (80.6%) and adaptation of insulin dosage (82.4%), an improved knowledge (82.2%), and a renewed motivation for the treatment (84.5%). Treatment success was significantly associated with baseline HbA1c, stability of motivation, frequency of blood glucose self-monitoring, control beliefs and change in subsequent outpatient care. CONCLUSIONS: The programme improved glycaemic control mainly as a result of a substantial reduction in the incidence of severe hypoglycaemia. Patients with persistent poor glycaemic control may benefit from structured follow-up care focusing on motivational aspects of self-management and psychosocial support. PMID- 11051283 TI - Plasma concentrations of VCAM-1 and ICAM-1 are elevated in patients with Type 1 diabetes mellitus with microalbuminuria and overt nephropathy. AB - AIMS: Elevated urinary albumin excretion is associated with macrovascular atherosclerotic complications in Type 1 diabetes mellitus. Adhesion molecules mediate leucocyte adhesion to the endothelium early in the atherosclerotic process. The present study tests the hypothesis that microalbuminuria and diabetic nephropathy are associated with elevated plasma concentrations of soluble vascular adhesion molecule (sVCAM)-1, soluble intercellular adhesion molecule (sICAM)-1, and soluble E-selectin (sE-selectin) aiming to illustrate factors of potential pathogenetic relevance for the excess cardiovascular disease in diabetic patients with renal complications. METHODS: Soluble adhesion molecule concentrations were measured by enzyme-linked immunosorbent assays (ELISA) in healthy controls (n = 16) and in 59 Type 1 diabetic patients: group 1-patients with normoalbuminuria (n = 16); group 2-patients with microalbuminuria (n = 15); group 3-patients with macroalbuminuria and normal serum creatinine (n = 15), group 4-patients with macroalbuminuria and moderately elevated serum creatinine (n = 13). RESULTS: Plasma concentrations of sVCAM-1 and sICAM-1 were similar in healthy controls and normoalbuminuric Type 1 diabetic patients, but the concentrations were increased by the presence of microalbuminuria and overt nephropathy (P < 0.001 and P < 0.0001, ANOVA). Concentrations of sE-selectin did not differ between diabetic patients and controls. CONCLUSIONS: Plasma concentration of sICAM-1 is elevated in Type 1 diabetic patients with microalbuminuria and the concentrations of sICAM-1 as well as sVCAM-1 are elevated in patients with macroalbuminuria and normal s-creatinine. The elevated plasma concentrations of these soluble adhesion molecule concentrations in patients with renal complication can be of pathogenetic importance for the development of atherosclerosis and plasma soluble adhesion molecule concentrations may provide additional information on cardiovascular risk. PMID- 11051284 TI - Vascular endothelial growth factor (VEGF) in children, adolescents and young adults with Type 1 diabetes mellitus: relation to glycaemic control and microvascular complications. AB - AIMS: To evaluate serum levels of vascular endothelial growth factor (VEGF) in a large group of children, adolescents and young adults with Type 1 diabetes mellitus to investigate whether increased VEGF concentrations are associated with long-term glycaemic control and microvascular complications. METHODS: The study involved 196 patients with Type 1 diabetes mellitus (age range 2-24 years, onset of diabetes before the age of 12 years, duration of disease longer than 2 years), without clinical and laboratory signs of microvascular complications; they were divided into three groups (group 1 - n = 37, age < 6 years; group 2 - n = 71, age 6-12 years; group 3 - n = 88, age > 12 years). Fifty-three adolescents and young adults (age 16.1-29.7) with different grades of diabetic retinopathy and microalbuminuria were also selected (group 4). A total of 223 healthy controls were matched for age and sex with each group of patients with diabetes mellitus. RESULTS: VEGF serum levels were significantly increased in pre-school and pre pubertal children with diabetes as well as in pubertal patients compared to controls. VEGF concentrations were markedly increased in adolescents and young adults with microvascular complications compared with both healthy controls and diabetic patients without retinopathy or nephropathy. Multivariate analysis showed that elevation of VEGF in serum was an independent correlate of complications. One-year mean HbA1c values were significantly correlated with VEGF concentrations (r = 0.372; P < 0.01). Children with HbA1c levels greater than 10% had significantly higher VEGF concentrations when compared with matched patients whose HbA1c levels were lower than 10%. In poorly controlled diabetic children (HbA1c > 10%), long-term (2 years) improvement of glycaemic control (aiming at HbA1c < 7%) resulted in a significant reduction of VEGF levels. CONCLUSIONS: VEGF serum concentrations are increased in prepubertal and pubertal children with diabetes. Glycaemic control influences VEGF serum levels. Severity of microvascular complications is associated with marked increase of VEGF concentrations in the serum of these patients. PMID- 11051285 TI - The glycaemic index of a range of gluten-free foods. AB - AIMS: The number of people with both diabetes and coeliac disease is increasing. This study examined the effect of gluten-free, as opposed to gluten-replete carbohydrate containing foods, on post-prandial blood glucose concentrations. METHODS: The glycaemic index of six commonly used gluten-free carbohydrates are reported and compared with published figures for similar non-gluten-free products. RESULTS: The results indicate that the glycaemic index of gluten-free and gluten containing foods are similar. CONCLUSIONS: The inclusion of gluten free foods in the diet of diabetic individuals with coeliac disease should not compromise glycaemic control. PMID- 11051286 TI - Assessment of the non-HLA-DR-DQ contribution to IDDM1 in British Caucasian families: analysis of LMP7 polymorphisms. AB - AIMS: Whilst HLA-DRB1 and HLA-DQ alleles contribute to IDDM1, the major determinant of genetic susceptibility to Type 1 diabetes mellitus, other major histocompatibility complex (MHC)-encoded genes may also be involved. The LMP7 (large multifunctional proteasome 7) gene is a potential candidate. The aim of this study was to assess whether LMP7 confers susceptibility to Type 1 diabetes independently of linkage disequilibrium with HLA-DRB1 and HLA-DQ. METHODS: The diallelic LMP7 polymorphism (LMP7*A or *B) was determined in 142 multiplex families from the British Diabetic Association Warren Repository. At least one parent was heterozygous for LMP7 in 112 families and these were informative for calculation of the statistic Tsp. This gives a valid chi2 test of the null hypothesis of no association or no linkage. RESULTS: An excess of transmissions of LMP7*A was observed from parents to affected offspring and the Tsp statistic was significant for association in the presence of linkage. LMP7*A was in positive, and LMP7*B in negative, linkage disequilibrium with the HLA-DRB1*03 DQ2, DRB1*04-DQ8 (group of all DRB1*04 subtypes), DRB1*0401-DQ8 and DRB1*0404-DQ8 haplotypes, although the linkage disequilibrium coefficient (delta) value was not statistically significant for DRB1*0404-DQ8. Analysis of HLA-DR-DQ-LMP7 haplotypes and Tsp analysis of HLA-matched-homozygous parents showed no association between LMP7 alleles and Type I diabetes independent of linkage disequilibrium with HLA-DR-DQ haplotypes associated with increased risk of disease. A contribution of LMP7 alleles to susceptibility to Type 1 diabetes in subjects with low-risk HLA-DR-DQ haplotypes could not be excluded. CONCLUSIONS: LMP7 alleles do not contribute to genetic susceptibility to Type 1 diabetes in subjects with high-risk-associated HLA-DR-DQ haplotypes. PMID- 11051287 TI - Autoantibodies to glutamic acid decarboxylase in diabetic patients from a multi ethnic Australian community: the Fremantle Diabetes Study. AB - AIMS: To investigate ethnic/racial differences in the prevalence of serum antibodies to glutamic acid decarboxylase (GADA) and ICA512/IA-2 in diabetic patients from a large, urban community. METHODS: A cross-sectional sample of 1,381 diabetic patients aged 11-98 years, representing 61% of those identified in a postcode-defined population base of 120,097 people were studied. Diabetes was classified on clinical grounds. Serum GADA and anti-ICA512/IA-2 were measured by radioimmunoprecipitation assay. RESULTS: Anglo-Celts formed 62% of the sample, southern Europeans 18%, other Europeans 8% and Asians 3%. GADA prevalence in Type 1 and Type 2 diabetes mellitus was 46.0% and 4.2%, respectively, amongst Anglo Celts and 22.2% and 1.7% in southern Europeans. The prevalence of anti-ICA512/IA 2 in Type 1 diabetes was 17.4% and, in a sample of 233 patients with Type 2 diabetes, 0.8%. GADA-positive Type 2 patients had a lower body mass index and greater glycosylated haemoglobin, and were more likely to be taking insulin, than GADA-negative Type 2 diabetic subjects (P < 0.05), consistent with the phentoype of latent autoimmune diabetes of adults (LADA). In both Type 1 and Type 2 diabetes, there was a strong inverse association between GADA and serum triglycerides (P < 0.001). CONCLUSIONS: The relatively low GADA prevalence in Anglo-Celt patients with Type 1 diabetes is a feature of this community-based study and suggests that GADA levels do fall with time, given the older age of the sample and a relatively long period between diagnosis and sampling. Southern Europeans had an even lower GADA prevalence, regardless of diabetes type. Variations in GADA frequency in diabetic patients of differing European ethnicity has implications for clinical management and healthcare planning. PMID- 11051288 TI - Incidence and predictors of drug-treated diabetes in elderly French subjects. The PAQUID Epidemiological Survey. AB - AIMS: To estimate the incidence and predictors of drug-treated diabetes in elderly subjects. METHODS: The PAQUID epidemiological survey, a population-based study, has followed up 3,777 subjects older than 65 years since 1988. At each visit (baseline, 1, 3, 5 and 8 years), treatment regimen was used to identify new drug-treated diabetic subjects. Potential predictors of drug-treated diabetes were collected during the baseline visit (body mass index (BMI), educational level, cigarette smoking and wine consumption, physical activity, depressive symptomatology, subjective health, treatment, and hypertension) and analysed by using a multivariate backward stepwise regression Cox model with delayed entry. RESULTS: The prevalence rate of drug-treated diabetes was 7.5% at baseline and 7.1% after 8 years' follow-up. The incidence rate of drug-treated diabetes was 3.8/1,000 person-years, 5.9/1,000 person-years in men and 2.4/1,000 person-years in women, with no significant variation according to age group. Male sex (relative risk (RR) 2.4, 95% confidence interval (CI) 1.4-4.0, P < 0.001, attributable risk (AR) 0.36), elevated BMI (for one point increase, RR 1.1, 95% CI 1.1-1.1, P < 0.001, > or = 25 vs. < 25, RR 2.1, 95% CI 1.2-3.5, AR 0.33), thiazide diuretics used alone (RR 5.9, 95% CI 1.8-19.6, P = 0.02), and poorer subjective health ('the same' vs. 'better' RR 1.8, 95% CI 1.0-3.1, P = 0.04; 'worse' vs. 'better' RR 2.3, 95% CI 0.9-5.7, P = 0.06) were independent predictors of drug-treated diabetes in this population. CONCLUSIONS: In older French individuals, men seem to be particularly exposed to drug-treated diabetes although being overweight was found to be a strong predictor as in younger populations. PMID- 11051289 TI - A case of hand ulceration in the diabetic foot clinic--a reminder of hand neuropathy in 'at risk' patients. AB - BACKGROUND: A case of hand ulceration in a diabetic patient with known lower extremity complications is presented. Although often asymptomatic, quantitative testing in patients indicates reduced hand sensation in patients with lower extremity neuropathy. Hand neuropathy may occasionally lead to anaesthetic injuries, particularly in certain 'manual' occupations, as seen in our patient. CONCLUSIONS: Education on hand care is virtually nonexistent in most clinic settings, and our case highlights the need for more awareness on this potentially troublesome complication. PMID- 11051290 TI - Chromium supplementation improves insulin resistance in patients with Type 2 diabetes mellitus. PMID- 11051291 TI - Heterogeneity in the clinical course of patients with Type 2 diabetes on dialysis -the need for different preventative strategies. PMID- 11051292 TI - Endothelial dysfunction in Type 1 diabetes mellitus: methodological considerations. PMID- 11051293 TI - Viral meningitis. AB - Enteroviruses account for 85 to 95% of all cases of aseptic meningitis, but the arboviruses and herpes simplex virus are also important etiologic agents. Mumps, lymphocytic choriomeningitis virus, herpes zoster, human herpesvirus type 6, and influenza viruses are rare causes of meningitis. The virology, pathogenesis, epidemiology, clinical manifestations, diagnostic studies, and established and potential antiviral therapies for viral meningitis are discussed. A differential diagnosis of the aseptic meningitis syndrome is provided. PMID- 11051294 TI - Acute bacterial meningitis. AB - In the past 10 years the epidemiology of bacterial meningitis has changed, with a decreased incidence of meningitis caused by Haemophilus influenzae and an increasing incidence of meningitis caused by penicillin- and cephalosporin resistant strains of Streptococcus pneumoniae. Meningococcal meningitis has become an increasing threat to college students. Successful outcome from meningitis requires not only eradication of the bacterial pathogen but also management of the neurological complications of raised intracranial pressure, stroke, and seizure activity. In this article, the pathophysiology, etiology, clinical presentation, differential diagnosis, and management of acute bacterial meningitis are reviewed. The present recommendations for the use of dexamethasone in the treatment of this infection, the use of chemoprophylaxis, and the indications for vaccinations are included. PMID- 11051295 TI - Fungal meningitis. AB - Fungi provide many benefits to humans. However, some of these fungi have the ability to become human pathogens. All the major fungal pathogens can produce meningitis. From the common cryptococcal meningitis to the rare fungal meningitis caused by a dimorphic or filamentous fungus, medical issues are discussed in this review on a fungus-specific basis. Both primary (Cryptococcus, Blastomyces, Histoplasma, Coccidioides, and other dimorphic fungi) and secondary (Aspergillus, Candida, and a series of molds) fungal pathogens can produce life-threatening central nervous system infections. These infections require immediate and precise diagnosis and carefully selected management strategies to optimize outcomes. In this review, we examine the epidemiology, pathogenesis, clinical manifestations, and treatment for fungal meningitis in all the major fungal groups. PMID- 11051296 TI - Encephalitis in the 21 st century. AB - As the 21st century begins, several outbreaks of encephalitis have been reported. An examination of these outbreaks brings into focus important epidemiological developments. Specifically, urbanization and encroachment on natural environments, the ease of world travel, and global trade can lead to spread of vectors and viruses from the developing world to the developed world. This review focuses on two recent epidemics of encephalitis: West Nile virus encephalitis in the eastern United States and Nipah virus encephalitis in Malaysia and Singapore. These examples demonstrate spread of a known viral agent from an endemic area to an area in which it had not previously been found and identification of a new viral agent. Infectious diseases in the developed world once considered "exotic" are now potential threats to all patients. PMID- 11051297 TI - Mycobacterium tuberculosis meningitis and other etiologies of the aseptic meningitis syndrome. AB - Mycobacterium tuberculosis is one of the most common infectious agents in the world. It causes an insidious form of meningitis characterized by headache, low grade fever, stiff neck and cranial nerve palsies, and an acute meningoencephalitis characterized by coma, raised intracranial pressure, seizures, and focal neurological deficits. This review focuses on the diagnosis and therapy of the insidious form of tuberculous meningitis and discusses the differential diagnosis of infectious and noninfectious etiologies of the aseptic meningitis syndrome. PMID- 11051298 TI - The prion diseases. AB - The prion diseases constitute an unusual group of neurodegenerative disorders. Although they are similar in many ways to other more common diseases, such as Alzheimer disease and amyotrophic lateral sclerosis, they are set apart on the basis of their transmissible nature. In addition to the unique feature of transmissibility, the prion diseases demonstrate that the expression of diverse disease phenotypes is possible from a common etiologic factor. This review provides the reader with a basic understanding of the nature of prions and highlights the clinical and pathologic features of these fascinating diseases. PMID- 11051299 TI - Brain abscess. AB - The epidemiology of brain abscess has changed with the increasing incidence of this infection in immunocompromised patients, particularly solid organ and bone marrow transplant recipients, and the decreasing incidence of brain abscess related to sinusitis and otitis. A number of new neuroimaging modalities, including single photon emission computed tomography, positron emission tomography, perfusion magnetic resonance imaging, and magnetic resonance spectroscopy, provide an initial noninvasive approach to diagnosis. The recommendations for the management of intracranial mass lesions in human immunodeficiency virus-infected individuals has changed as the incidence of toxoplasmic encephalitis has decreased with the use of trimethoprim sulfamethoxazole prophylaxis. The epidemiology, pathogenesis, microbiology, clinical presentation, diagnosis, treatment and prognosis of brain abscess in the beginning of the 21 st century are provided in this review. PMID- 11051300 TI - Listeria and atypical presentations of Listeria in the central nervous system. AB - Listeria monocytogenes infection of the central nervous system is often not recognized and treated appropriately in the crucial early stages of the disease. Most consider patients with underlying disease or immunocompromised states to be at risk, although healthy individuals may present with a neurologic syndrome caused by L. monocytogenes. Earlier suspicion and treatment remains our best means of reducing the morbidity and high mortality rate of this treatable disease. In addition to meningitis and meningoencephalitis, infection of the brainstem (rhomboencephalitis) is challenging to recognize and therefore initiate appropriate early therapy. Cerebritis and abscess can also occur. Furthermore, empirical therapy for meningitis or the other manifestations of nervous system involvement is often inadequate. This review addresses the clinical microbiology, pathogenesis, spectrum of neurological involvement, and treatment of central nervous system infection related to L. monocytogenes. PMID- 11051301 TI - Controversies in neurological infectious diseases. AB - The past several years have seen major advances in our understanding of neurological infectious diseases, their diagnosis, and their treatment. Along with these advances, however, new information about infectious agents and new therapeutic options have also introduced both uncertainty and controversy in the approach and management of patients with diseases of the central nervous system. Here, we discuss six such areas: the long-term efficacy of HAART therapy in treatment of HIV infection; the role of viral infection in chronic fatigue syndrome; Rasmussen's encephalitis as an infectious or autoimmune disease; the spectrum of neurological diseases caused by rickettsial infection; the role of Mycoplasma pneumoniae in human central nervous system disease; and the possible association of Chlamydia pneumoniae and human herpesvirus 6 with multiple sclerosis. PMID- 11051302 TI - Diagnosis and management of systemic infections and fever in neurological patients. AB - The hospital course of a patient with a neurological disorder may be complicated by the development of fever, urinary tract infection (UTI), bacteremia, pneumonia, diarrhea, pressure sores, or intravenous line infections. An approach to each of these is provided in this review as well as recommendations for empirical therapy of catheter-related UTIs, intravenous line infections, and aspiration pneumonia. PMID- 11051303 TI - Suggested dosing regimens and adverse effects of antimicrobials used in central nervous system infections. PMID- 11051304 TI - Mathematical models for predicting G-duration tolerances. AB - Mathematical models that predict fatigue-based G-duration tolerances for relaxed and straining subjects are developed and validated using published data. These models are based on regression analysis calculations using published G-duration tolerance data of relaxed subjects exposed to 3-5 G and subjects exposed to 6-9 G using an anti-G suit and performing the anti-G straining maneuver. These G duration models are derived from published G-level tolerance models based on intravascular hydrostatic pressures and physiologic responses to maximum voluntary contractions (MVC%). Included in the validation of these models are the baroreceptor and muscle contraction cardiovascular reflexes that support arterial BP. A basic energy pool that supports a G-duration of 140 s for G exposures > 5 G is theorized. Because of the long duration of sustained G exposures in these models, the physiologic dynamics involved in predicting straining G-duration tolerances, are identified and validated using different time periods, i.e., Phases I and II. These models, based on sustained G exposures to a constant G level are also applicable to exposures of variable G levels known as simulated aerial combat maneuver (SACM) G-profile tolerances. G-duration tolerances > 9 G are predicted using these models for subjects using reclined-seat backs and positive pressure breathing. PMID- 11051305 TI - Effect of normobaric hypoxia on sound localization. AB - BACKGROUND: Three-dimensional (3D) audio displays have considerable potential for conveying spatial information in the aviation environment. How reliably these displays will function in that environment will depend in part on the extent to which sound localization is affected by hypoxia. Many aircraft systems operators are routinely exposed to mild hypoxia, and all are at risk of exposure to more severe hypoxia. METHODS: We have examined hypoxia's effect on localization by measuring the localization performance of four subjects exposed to simulated altitudes of 0, 1200, 2400 and 3700 m above sea level for about 30 min. Sounds to be localized were presented from a free-field source at locations covering 360 degrees of azimuth and extending from -47.6 to + 80 degrees elevation. RESULTS: Localization performance was statistically indistinguishable across the altitudes tested. Average localization errors ranged from 12.6 degrees +/- 0.7 SE at 2400 m to 14 degrees +/- 0.84 SE at 0 m. CONCLUSION: The finding that hypoxia induced by exposure to simulated altitudes as high as 3700 m has no effect of sound localization is encouraging with respect to the continued development of 3D audio displays for use in the aviation environment. PMID- 11051306 TI - Effect of seating, vision and direction of horizontal oscillation on motion sickness. AB - BACKGROUND: Low frequency horizontal oscillation can cause motion sickness in some forms of transport, but the influence of the characteristics of the motion and the visual and postural conditions of the body on sickness are not known. HYPOTHESES: It was hypothesised that body position, vision and direction of motion will have an effect on motion sickness. METHOD: There were 72 seated subjects who were exposed to horizontal oscillation at 0.25 Hz, 0.7 ms(-2) r.m.s. (peak-to-peak displacement of 0.8 m) for up to 30 min while in 1 of 6 conditions. Three conditions involved fore-and-aft motion and three involved lateral motion. For motion in each axis, subjects sat within a closed cabin with either: a) a high backrest with their eyes open; b) a low backrest with their eyes open; or c) a low backrest with their eyes closed and blindfolded. Subjects provided ratings of their motion sickness symptoms at 1-min intervals during the 30-min exposures. RESULTS: The most nauseogenic stimulus was fore-and-aft motion with a low backrest and the eyes open. Self-ratings of motion sickness susceptibility provided by subjects before participating in the experiment were positively correlated with their illness ratings during the experiment. CONCLUSIONS: Restraint to the upper body during exposure to horizontal acceleration may reduce the susceptibility to motion sickness caused by horizontal oscillation. The relative nauseogenicity of fore-and-aft and lateral oscillation depends on the support given to the upper body. In the conditions of the experiment the effects of the postural support given to the subjects and their prior susceptibility to motion sickness were greater than any effect of the visual conditions. PMID- 11051307 TI - Promethazine affects optokinetic but not vestibular responses in monkeys. AB - BACKGROUND: Promethazine is used to treat motion sickness including Space Adaptation Syndrome, but there is incomplete information about how it affects vestibular and optokinetic responses. METHODS: Vestibular and optokinetic nystagmus, recorded with eye coils, were characterized in monkeys after administration of promethazine at dosages approximately equivalent to those used by humans in space. RESULTS: The initial increase of horizontal eye velocity during optokinetic nystagmus (OKN) was reduced after receiving the drug. Consequently, it took a longer time for eye velocity to rise to 60% of steady state value, the normal initial jump in eye velocity. Steady state OKN, maximum gains of optokinetic after-nystagmus (OKAN) and OKAN falling time constants were unaffected. The gains and time constants of the horizontal, vertical and roll angular vestibulo-ocular reflex (aVOR), the amplitude and velocity of saccades, and ocular counter-rolling (OCR), induced by off-vertical axis rotation (OVAR) were unaffected by promethazine. A two-component optokinetic model simulated the data simply by reducing the gain of the initial (rapid) component of OKN. A reduction in coupling between a non-linear element and the velocity storage integrator was required to simulate some vertical OKN data. CONCLUSIONS: Promethazine reduces the gain of the direct visual-oculomotor pathway in monkeys. It has little effect on saccades, the gain and time constant of the aVOR and the low frequency linear vestibulo-ocular reflex (IVOR), which orients the eyes during ocular counterrolling. The optokinetic deficit is consistent with reported reduction in ocular pursuit and VOR suppression after promethazine in humans. PMID- 11051308 TI - Promethazine as a motion sickness treatment: impact on human performance and mood states. AB - PURPOSE: Intramuscular (i.m.) injections of promethazine in 25 mg or 50 mg dosages are commonly used to treat space motion sickness in astronauts. The present study examined the effects of i.m. injections of promethazine on performance, mood states, and motion sickness in humans. METHODS: Subjects were 12 men, mean age 36 + 3.1, who participated in 1 training day and 3 treatment conditions: a 25-mg injection of promethazine, a 50-mg injection of promethazine, and a placebo injection of sterile saline. Each condition, scheduled at 7-d intervals, required an 8-10-h day in which subjects were tested on 12 performance tasks, and were given a rotating chair motion sickness test. On the training day subjects were trained on each task to establish stability and proficiency. Treatment conditions were counterbalanced and a double-blind procedure was used to administer the medication or placebo. RESULTS: Statistically significant decrements in performance were observed for both dosages of promethazine as compared with the placebo. Performance decrements were associated with mean blood alcohol dose equivalency levels of 0.085% for 25 mg and 0.137% for 50 mg doses. Mood scale results showed significant changes in individual subjective experiences with maximum deterioration in the arousal state and fatigue level. Only the 25-mg dosage significantly increased motion sickness tolerance when compared with the placebo. CONCLUSIONS: These data suggest that effective doses of promethazine currently used to counteract motion sickness in astronauts may significantly impair task components of their operational performance. PMID- 11051309 TI - Effectiveness of an expert system for astronaut assistance on a sleep experiment. AB - BACKGROUND: Principal Investigator-in-a-Box ([PI]) is an expert system designed to train and assist astronauts with the performance of an experiment outside their field of expertise, particularly when contact with the Principal Investigators on the ground is limited or impossible. In the current case, [PI] was designed to assist with the calibration and troubleshooting procedures of the Neurolab Sleep and Respiration Experiment. [PI] displays physiological signals in real time during the pre-sleep instrumentation period, alerts the astronauts when a poor signal quality is detected, and displays steps to improve quality. METHODS: Two studies are presented in this paper. In the first study 12 subjects monitored a set of prerecorded physiological signals and attempted to identify any signal artifacts appearing on the computer screen. Every subject performed the experiment twice, once with the assistance of [PI] and once without. The second part of this study focuses on the postflight analysis of the data gathered from the Neurolab Mission. After replaying the physiological signals on the ground, the frequency of correct [PI] alerts and false alarms (i.e., incorrect diagnoses by the expert system) was determined in order to assess the robustness and accuracy of the rules. CONCLUSIONS: Results of the ground study indicated a beneficial effect of [PI] and training in reducing anomaly detection time and the number of undetected anomalies. For the in-flight performance, excluding the saturated signals, the expert system had an 84% detection accuracy, and the questionnaires filled out by the astronauts showed positive crew reactions to the expert system. PMID- 11051310 TI - Cerebrovascular responses during lower body negative pressure-induced presyncope. AB - BACKGROUND: Reduced orthostatic tolerance is commonly observed after spaceflight, occasionally causing presyncopal symptoms which may be due to low cerebral blood flow (CBF). It has been suggested that CBF decreases in early stages of exposure to orthostatic stress. The purpose of this study was to investigate cerebrovascular responses during presyncope induced by lower body negative pressure (LBNP). HYPOTHESIS: Although CBF decreases during LBNP exposure, blood pressure (BP) or heart rate (HR) contributes more to induce presyncopal conditions. METHODS: Eight healthy male volunteers were exposed to LBNP in steps of 10 mm Hg every 3 min until presyncopal symptoms were detected. Electrocardiogram (ECG) was monitored continuously and arterial BP was measured by arterial tonometry. CBF velocity at the middle cerebral artery was measured by transcranial Doppler sonography (TCD). Cerebral tissue oxygenation was detected using near-infrared spectroscopy (NIRS). We focused our investigation on the data obtained during the final 2 min before the presyncopal endpoint. RESULTS: BP gradually decreased from 2 min to 10 s before the endpoint, and fell more rapidly during the final 10 s. HR did not change significantly during presyncope. CBF velocity did not change significantly, while cerebral tissue oxygenation decreased prior to the presyncopal endpoint in concert with BP. Our results suggest that CBF is maintained in the middle cerebral artery during presyncope, while BP decreases rapidly. CONCLUSIONS: Cerebrovascular hemodynamics are relatively well maintained while arterial hypotension occurs just prior to syncope. PMID- 11051311 TI - Histologic changes in the adrenal cortex from young rats following spaceflight. AB - BACKGROUND: Potential stresses associated with spaceflight include microgravity, acceleration and deceleration forces, a crowded environment and re-adaptation to normal gravity after landing. HYPOTHESIS: We hypothesized that spaceflight would result in histological changes in the adrenal glands of young rats. METHODS: Six week old male rats were group-housed in an Animal Enclosure Module (AEM) for a 17 d shuttle flight (STS-78). Ground-based controls included a baseline group, an AEM-housed group and a vivarium group. Adrenal glands were collected from 4-6 hours after flight, fixed, embedded in plastic and sections prepared for light microscopy. RESULTS: The adrenals from the baseline and vivarium groups had normal histological features. Some changes in the adrenal cortices from the ground-based AEM group included greater parenchymal cord-like formation. The adrenal weights and width of the zona fasciculata were greater in the flight group than the controls. There were also increased parenchymal cord-like formation with better demarcation of the vascular sinusoids in the zona reticularis and zona fasciculata, greater depletion of cytoplasmic lipid vacuoles, and an increased nuclear volume of the cells in the zona fasciculata when compared with the controls. CONCLUSIONS: The adrenal changes in the ground based AEM animals may be attributed to the confined space in the AEM. The adrenal enlargement and the histological changes observed in the flight animals may be attributed to spaceflight and possibly re-entry in addition to possible confinement stress in the AEM. PMID- 11051312 TI - Intraocular pressure and acclimatization to 4300 M altitude. AB - BACKGROUND: Studies were conducted to determine the effect of altitude exposure on intraocular pressure (IOP) and any relationship with the severity of acute mountain sickness (AMS). HYPOTHESES: a) IOP is decreased during exposure to 4300 m altitude; b) there is a positive correlation between IOP and AMS; and c) there is a correlation between changes in urinary catecholamines and IOP. METHODS: IOP (noncontact tonometry) was measured in 11 resting males during acute simulated altitude (446 mmHg, < 2 h, hypobaric chamber), during altitude acclimatization (15 d at 4300 m), and in 6 of the 11 volunteers during re-exposure in the chamber after 8 d at sea level (Study A). In a second study (Study B) of 12 females, IOP (contact tonometry) and 24-h urinary catecholamines were measured during a 50-h chamber exposure (446 mmHg). AMS severity was assessed using the Environmental Symptoms Questionnaire (ESQ-C). RESULTS: IOP decreased 25% after 2 d at altitude and returned toward sea level values by 15 d (Study A). IOP was reduced 13% after 5 h of exposure followed by return toward sea level values (Study B). Significant correlation was found between the sea level IOP and ESQ-C (Study A); significant correlation was found between the reduction in IOP and the ESQ-C and urinary epinephrine concentrations (Study B). CONCLUSIONS: Altitude exposure resulted in a reduction in IOP that occurred within hours and recovered during acclimatization. This reduction may be related to increases in epinephrine concentration. Measurement of IOP before and during altitude exposure may provide an objective method of assessing an individual's response to hypoxic stress. PMID- 11051313 TI - Decompression scenarios in a new underground transportation system. AB - BACKGROUND: The risks of a public exposure to a sudden decompression, until now, have been related to civil aviation and, at a lesser extent, to diving activities. However, engineers are currently planning the use of low pressure environments for underground transportation. This method has been proposed for the future Swissmetro, a high-speed underground train designed for inter-urban linking in Switzerland. HYPOTHESIS: The use of a low pressure environment in an underground public transportation system must be considered carefully regarding the decompression risks. Indeed, due to the enclosed environment, both decompression kinetics and safety measures may differ from aviation decompression cases. METHOD: A theoretical study of decompression risks has been conducted at an early stage of the Swissmetro project. A three-compartment theoretical model, based on the physics of fluids, has been implemented with flow processing software (Ithink 5.0). Simulations have been conducted in order to analyze "decompression scenarios" for a wide range of parameters, relevant in the context of the Swissmetro main study. RESULTS: Simulation results cover a wide range from slow to explosive decompression, depending on the simulation parameters. Not surprisingly, the leaking orifice area has a tremendous impact on barotraumatic effects, while the tunnel pressure may significantly affect both hypoxic and barotraumatic effects. Calculations have also shown that reducing the free space around the vehicle may mitigate significantly an accidental decompression. CONCLUSION: Numeric simulations are relevant to assess decompression risks in the future Swissmetro system. The decompression model has proven to be useful in assisting both design choices and safety management. PMID- 11051314 TI - Ejection from an aircraft following photorefractive keratectomy: a case report. AB - A 32-yr-old active duty United States Navy Lieutenant Naval Flight Officer (NFO), 6 mo status post photorefractive keratectomy (PRK) in both eyes performed as part of the first protocol to evaluate laser refractive surgery in aviators, ejected from a Navy S-3B Viking aircraft while performing field carrier landing practice. Neither the pilot nor the NFO suffered physical consequences as a result of the ejection. Coincidentally, the NFO had been examined just 1 wk before the mishap and was noted to have uncorrected visual acuity of 20/16 in each eye. Examination 1 wk following the mishap could not detect a change in vision. The NFO's vision status and his post-PRK status was not listed as a causal factor in the mishap. While this case report will not settle the debate regarding the appropriateness of refractive surgery in aviators, it does provide anecdotal evidence to support the safety of PRK in the aviation community. PMID- 11051315 TI - Laser pointers and aviation safety. AB - Laser pointers have been used by teachers and lecturers for years to highlight key areas on charts and screens during visual presentations. When used in a responsible manner, laser pointers are not considered to be hazardous. However, as the availability of such devices has increased, so have reports of their misuse. The Food and Drug Administration (FDA) issued a warning in December 1997 on the possibility of eye injury to children from handheld laser pointers. In October 1998, the American Academy of Ophthalmology upgraded an earlier caution to a warning, stating that laser pointers can be hazardous and should be kept away from children, after two reports of eye injuries involving young girls (age 11 and 13 yr). Of particular concern was the promotion of laser products as children's toys, such as those that can project cartoon figures and line drawings. Additionally, there have been reports involving the misuse of laser pointers (e.g., arrests made after police interpreted the red beam to be a laser sighted weapon, spectators aiming laser lights at athletes during sporting events, cars illuminated on highways, and numerous incidents involving the illumination of aircraft). This technical note discusses physiological effects of exposure from a laser pointer, the regulation and classification of commercial laser products, and how the misuse of these pointers is a possible threat to aviation safety. PMID- 11051316 TI - Aeromedical concerns of the air traffic controller: the forgotten second-class medical. PMID- 11051317 TI - FAA proposed rule for AEDs and inflight medical kits. PMID- 11051318 TI - FAA flight data link, operational at Oshkosh. PMID- 11051319 TI - Pattern of gonadotropin secretion and ultrasonographic evaluation of developmental changes in the testis of early and late maturing bull calves. AB - This was a study that retrospectively analyzed serum gonadotropin secretion and the ultrasonographic appearance of the testis during development in prepubertal bull calves to determine whether there were differences between early and late maturing bulls. Blood samples were taken every other week from 2 wk of age until puberty. Samples were also taken at 12 minute intervals for 12 hours at 4, 10, 20, 25, 30, 35, 40 and 45 wk of age. The GnRH treatment was administered 10 hours after the start of each period of frequent blood sampling. Bull calves fell into two distinctive groups, with one group maturing between 36.6 and 44.2 wk (n = 12) and the other between 46.4 and 48.9 wk of age (n = 8). In samples taken every other week mean serum LH concentrations were greater in early maturing bulls than in late maturing bulls at 12, 14 and 16 wk of age (P<0.05). In blood samples taken every 12 minutes for 10 hours early maturing bull calves had higher mean serum LH concentrations at 4 and 10 wk of age (P<0.05) and higher LH pulse frequency at 10 and 20 wk of age (P<0.05). Mean serum LH concentrations at 4, 10 and 40 wk of age and LH pulse frequency at 10 and 20 wk of age were negatively correlated with age at puberty in bull calves. Mean pixel units of the right and left testis were higher from 34 to 40 wk of age in early maturing than in late maturing animals (P<0.05). It seems possible that hormone measurements and ultrasonographic characteristics of the testes could be developed into powerful tools for studies on the regulation of reproductive development and may aid in the prediction of reproductive potential. PMID- 11051320 TI - Epithelial and stromal uterine cells cultured in vitro protect bovine sperm from hydrogen peroxide. AB - It is known that large amounts of leukocytes colonize the uterus, and that these leukocytes can produce considerable quantities of hydrogen peroxide (H2O2) and other reactive oxygen species that are toxic to sperm. It has been shown recently that oviductal fluid has a catalase that helps to maintain sperm motility. Therefore, the current experiment was performed to determine if a similar mechanism of protection exists against peroxides within uterine cells. Sperm motility and velocity were recorded after a 6h incubation in 1) conditioned media in the presence of endometrial cells, 2) conditioned media without endometrial cells, 3) control media (48h without cells) over endometrial cells, or 4) control media alone. All these treatments were performed in the presence or absence of added catalase. Conditioned media, endometrial cells and catalase had a significant positive effect on the maintenance of sperm motility and velocity. Addition of anti-catalase antibodies did not neutralize the beneficial effect of the conditioned media. However, the concentrations of aromatic amino acids, known substrates for sperm amino acid oxidase, were significantly lower in uterine conditioned media as compared to control medium. This reduction of aromatic amino acids was in correlation with reduced H2O2 production by sperm as estimated by chemiluminescence. These results suggest that epithelial and stromal uterine cells do not maintain sperm motility by secreting catalase in the conditioned media, but rather by reducing the levels of aromatic amino acids and thus of peroxides generated in the presence of spermatozoa. PMID- 11051321 TI - Early identification of neonates at risk: traits of newborn piglets with respect to survival. AB - Despite technological changes and improved management, piglet mortality remains a problem for both production and welfare. Most preweaning mortality occurs within the first 3 days after birth because of problems with adaptation and development. Thus, the purpose of our study was to determine the physiologic state of newborn pigs with respect to piglet survival. Data were collected from 1024 live-born piglets of 106 primiparous German Landrace sows to analyze relationships between farrowing traits, early postnatal vitality and blood chemistry, including immunity of piglets at birth. Surviving piglets were compared with those that died during the first 10 days of life. The survivors were significantly heavier at birth (P=0.001), were born earlier in the birth order (P=0.04), reached the udder and took in first colostral milk more quickly (P=0.001) and had a smaller drop in rectal temperature I h after birth (P=0.001) than dead. However, dead piglets had significantly higher blood levels of inorganic phosphorus (P=0.0001), calcium (P=0.04) and urea (P=0.05), but a lower concentration of alpha2 macroglobulin and lower lymphocyte proliferation indices in response to pokeweed mitogen (P=0.05). Models fitted for discrimination between survivors and piglets that died included, in addition to birth weight and litter size, the foraging behavior of neonates (time from birth to first suckle) and their thermoregulatory capacity (rectal temperature 1 h after birth) in the first experimental unit, as well as prenursing biochemical measures (inorganic phosphorus, calcium and glucose) in the second experimental unit. These ethophysiological and biochemical traits of early postnatal vitality are important determinants of maturity and development at birth. Hence, breeding programs and perinatal housing and feeding conditions should ensure a high physiological maturity to improve mortality rates of neonates. PMID- 11051322 TI - Changes in porcine oocyte germinal vesicle development as follicles approach preovulatory maturity. AB - This study was conducted to determine the distribution of oocytes in meiotic arrest as a function of follicle maturation, atresia status, and follicular fluid steroid concentrations. Oocytes (n = 138) from > or = 3 mm follicles were recovered from gilts (n = 3/d) on Days 1, 3, 5, and 7 of the follicular phase initiated by withdrawal of altrenogest treatment. They were fixed in 4% paraformaldehyde, stained with Hoechst 33342, and examined by laser scanning confocal microscopy using combined bright field Nomarski optics and ultraviolet laser illumination. The number of oocytes in complete meiotic arrest increased (P < 0.05) as a function of the stage of maturation from 29% on Day 1 to 79 and 67% on Days 3 and 5, respectively. Oocytes showing complete germinal vesicle breakdown (GVBD) were found only on Day 7 (24 to 36 h after the preovulatory LH surge). The distribution of GV stages on Days 1 to 5 did not differ between atretic (n = 27) and nonatretic follicles (n = 81). In nonatretic follicles, GV stage was inversely related to the concentration of estradiol on Day 7 and to the concentrations of progesterone and androstenedione (P < 0.05) on Days 5 and 7 indicating that meiotically arrested oocytes were likely to be found in follicles with highest levels of steroidogenesis. In conclusion, a large proportion of oocytes present in 3 to 5 mm follicles had begun GVBD. The follicles in the ovulatory cohort may be recruited or selected from preexisting 3 to 5 mm follicles, or younger population with oocytes that are in complete meiotic arrest. PMID- 11051323 TI - Prevalence and reproductive effects of Ureaplasma diversum in beef replacement heifers and the relationship to blood urea nitrogen level. AB - A systematic sample of replacement heifers from 5 herds underwent prebreeding vaginal swab cultures for Ureaplasma diversum. Heifers from three of the herds were subsequently sampled at pregnancy examination. Sampled heifers were given a vaginal lesion score (VLS), reproductive tract score (RTS) and body condition score (BCS), and peripheral blood was collected for serum blood urea nitrogen (BUN) estimation. Culture results revealed an overall prevalence of Ureaplasma diversum of 51% (87/171) at prebreeding and 65% (64/98) at pregnancy examination. Within herd prevalence ranged from 36% to 64% at prebreeding and 54% to 76% at pregnancy examination. Prevalence tended to differ between herds (P=0.08). At the prebreeding examination, heifers with a BCS of 5.5 or less were more likely to be culture positive than heifers with a BCS greater than 5.5 (p<0.05). No relationship was noted between BUN, VLS, RTS, or pregnancy status and prebreeding culture status. There was little variability among the heifers for any of these variables, with vaginal lesion scores generally being mild, RTS scores being high and BCS scores being moderate. At pregnancy examination, heifers that were culture negative tended to be more likely to be pregnant (odds 3.7, p=0.10) than culture positive heifers. PMID- 11051324 TI - The influence of ovarian activity and uterine involution determined by ultrasonography on subsequent reproductive performance of dairy cows. AB - The objective of this study was to test the hypothesis that a follicle >8 mm diameter in the ovary ipsilateral to the previously gravid uterine horn (PGUH), between 14 and 28 days postpartum, improves subsequent reproductive performance. Lactating Holstein-Friesian cows (n=284) in 3 commercial herds were examined using transrectal ultrasonography once between 14 and 28 days postpartum to determine associations between uterine and ovarian measurements and subsequent fertility. There were fewer cows with a corpus luteum in the ovary ipsilateral to the PGUH compared with the contralateral ovary (16.9% vs. 37.0%; P<0.001). In addition, in the ovary ipsilateral to the PGUH there were fewer follicles >5 mm diameter (mean +/- SEM; 0.69 +/- 0.06 vs. 1.02 +/- 0.06; P<0.001) and fewer animals with a follicle >8 mm diameter (26.1% vs. 49.6%; P<0.001). These differences between the ovaries ipsilateral or contralateral to the PGUH declined with increasing time between 14 and 28 days postpartum. The presence of a purulent vaginal discharge decreased the number of animals with a corpus luteum in the ovary contralateral to the PGUH (45/143 vs. 60/141; P<0.05), but not in the ovary ipsilateral to the PGUH. The presence of a follicle >8 mm diameter in the ovary ipsilateral to the PGUH was associated with a shorter calving to conception interval compared with animals without such a follicle (99.0 +/- 5.6 days, n=74, vs. 112.8 +/- 4.4 days, n=210; P<0.05). These observations raise an intriguing issue: how does this follicle affect subsequent fertility--does the follicle exert a local influence on the uterus, or vice versa? PMID- 11051325 TI - Effects of dominant follicle aspiration and treatment with recombinant bovine somatotropin (BST) on ovarian follicular development in nelore (Bos indicus) heifers. AB - Follicle ablation has been recognized as an efficient method of follicular wave synchronization. Treatment with recombinant bovine somatotropin (BST) has been shown to enhance follicular development in Bos taurus. This experiment assessed the effects of these treatments in Nelore (B. indicus) heifers. Eight cycling Nelore heifers were randomly assigned to 3 different treatments. On Day 2 of a synchronized cycle (Day 0 = day of ovulation), heifers assigned to Treatments 1 and 2 received 2 mL of saline, whereas heifers assigned to Treatment 3 received 320 mg of BST. On Day 5, the first-wave dominant follicle was ablated by ultrasound-guided transvaginal aspiration in heifers in Treatments 2 and 3, and all heifers received an injection of prostaglandin on Day 11. Aspiration of the dominant follicle advanced and synchronized (P < 0.05) the day of second-wave emergence (6.9 +/- 0.1 vs. 8.4 +/- 0.4) and the day of the pre-wave FSH peak (6.0 +/- 0.0 vs. 6.9 +/- 0.4), and increased FSH peak concentrations (381 +/- 21 vs. 292 +/- 30; pg/mL; P < 0.01). Recombinant bovine somatotropin treatment caused a two-fold increase in plasma insulin-like growth factor-I (IGF-I) concentrations (P < 0.001) and resulted in a 36% increase in the number of small follicles (<5 mm; P < 0.001) compared with saline-treated heifers. In summary, in agreement with previous reports on B. taurus, dominant follicle aspiration synchronized ovarian follicular development, and BST treatment increased peripheral concentrations of IGF-I in Nelore heifers. Recombinant bovine somatotropin also increased the number of small follicles, but this response appeared to be inferior to that reported for B. taurus. PMID- 11051326 TI - Factors affecting recovery and quality of oocytes for bovine embryo production in vitro using ovum pick-up technology. AB - In Experiment 1, different vacuum pressures (30, 50, 70 and 90 mm Hg) were used to aspirate 4156 bovine follicles in vitro, to assess their effect on flow rate and the recovery, morphology and blastocyst formation of the recovered oocytes. Cumulus oocyte complexes (COCs) were classified according to the morphology of the cumulus cells. Data were analyzed using Chi Square analysis. Overall recovery rate declined as the aspiration pressure increased above 50 mm Hg (P<0.05). The recovery rate of Grade 1 oocytes decreased significantly (P<0.05) as the vacuum pressure increased with a corresponding increase in the number of denuded oocytes recovered (P<0.05). The blastocyst yield, expressed as a percentage of recovered COCs decreased significantly as the aspiration pressure increased beyond 50 mm Hg (P<0.05). In Experiment 2, the holding media (hepes- or bicarbonate-buffered TCM 199) and holding time (1 h or 5 h) did not affect the blastocyst formation of the oocytes (P>0.05). In Experiment 3, it was found that individual culture of the oocyte during fertilization or culture had a detrimental effect on the oocytes blastocyst formation (8.8% to 16% blastocyst yield on Day 8) when compared to control (31.3%). In Experiment 4, groups of 5, 10 and 25 oocytes were compared with singly cultured oocytes. There were no significant differences (P<0.05) in the blastocyst formation rate among groups of 5, 10, or 25 oocytes, but there was a significant difference between oocytes processed in groups and those processed individually. In Experiment 5, when groups of 10 oocytes were cultured in different drop sizes, there was no significant difference in cleavage rates between oocytes cultured in 100 microL (85.0%, n = 280) and in 10 microL (86.8%, n = 280) of media, but culture in 50 microL (79.3%, n = 280) resulted in cleavage rates significantly lower (P<0.05) than culture in 10 microL drops. There was no significant difference in the blastocyst formation. However there was a significant difference (P<0.05) in cell numbers of Day 8 blastocvsts, with oocytes cultured in 100 microL drops having significantly lower cell counts than oocytes cultured in 50 or 10 microL drops. PMID- 11051327 TI - Effect of dose and day of treatment on uterine response to oxytocin in mares. AB - To determine the effect of dose and day of oxytocin treatment on intrauterine pressure, 6 normal mares were treated with 10 or 25 IU oxytocin 2 days before ovulation, on the day of ovulation and 2 days after ovulation. Intrauterine pressure (IUP) was measured using micro-tip-catheters (one placed intrauterine, a second and third serving as reference sensors in the vagina and external to the mare) and transmitted by telemetry for 30 min to establish a baseline before saline was administered, iv, and for an additional 30 min after saline administration. Oxytocin was then given, iv, and IUP was recorded for 60 min. No change in IUP was observed after saline injection. The administration of both 10 (n=16) and 25 (n=10) IU oxytocin induced a response (P<0.01). The intensity of response depended on the day of administration (P<0.01) and the dose of oxytocin (P<0.001). The variation of response was significantly greater after 10 IU oxytocin (CV 15.78%) compared with 25 IU oxytocin (CV 6.42%). The uterine response was greatest on Day 2 prior to ovulation and lowest on Day 2 after ovulation. The response was negatively correlated to increasing plasma progesterone (10 IU oxytocin: r = -0.435, 25 IU oxytocin: r = -0.265). There was no correlation between the uterine response and plasma estradiol-17beta concentration (P<0.01). In conclusion the results of this study show that oxytocin administration to mares before ovulation provides a greater response than after ovulation. A decline in the intensity of response after ovulation can be compensated for with a higher dose of oxytocin. Furthermore, the use of the multiple catheter technique is an effective method for assessing changes in uterine pressure. PMID- 11051328 TI - The induction of parturition in the bitch using sodium cloprostenol. AB - The objectives of this studies were to determine a continuous low-dose treatment regimen for the administration of sodium cloprostenol to the bitch that did not cause polydipsia, and whether this treatment would induce normal and timed parturition in bitches during late pregnancy. Non-pregnant greyhound bitches (n=18) received sodium cloprostenol subcutaneously, via a miniosmotic pump, at dose rates of 0.875 to 4.5 microg/kg/24 h, for 7 days (Days 0 to 7). Daily water intake was measured from Day -2 to Day 9. Polydipsia was observed in bitches treated with the higher dose rates but not in bitches treated with the lowest dose rate of 0.875 microg/kg/24 h. In the second experiment, pregnant greyhound bitches received sodium cloprostenol at dose rates of 1 (n=4), 2 (n=1) and 3 microg/kg/24 h (n=1), on Day 57 of pregnancy. Polydipsia was observed in bitches treated at the higher dose rates of 2 and 3 microg/kg/24 h, but not in the bitches treated at the lower dose rate of 1 microg/kg/24 h. These treatments resulted in the successful induction of parturition. Parturition was associated with a decrease in plasma progesterone concentrations, a reduction in body temperature, and an increase in plasma concentrations of 13,14-dihydro-15-keto prostaglandin F2alpha. The first puppy was born 37.7 +/- 2.9 h after the start of treatment (range 28 to 46 h). The duration of whelping was approximately 15.7 +/- 2.2 h (range 10 to 24 h). The litter size was 9.2 +/- 0.8 pups (range 6 to 12 pups), and the puppy survival rate was 6.0 +/- 0.8 per litter (range 4 to 9 pups). This study demonstrated that the administration of sodium cloprostenol in continuous low dose for 24 h is an effective treatment for the induction of parturition in bitches during late pregnancy. This treatment resulted in the birth of healthy pups, with minimal or no side effects to the bitch. PMID- 11051329 TI - Sperm survival during short-term storage and after cryopreservation of semen from striped trumpeter (Latris lineata). AB - Methods of short-term storage and cryopreservation were examined for semen from striped trumpeter (Latris lineata). For fresh semen at 18 degrees C, the percentage of motile sperm declined rapidly from over 80% immediately after activation with sea water to less than 2% within 9 min after activation. The motility after activation of undiluted fresh sperm stored at 5 degrees C was maintained for two days and then declined markedly so that by the eighth day, sperm were mostly immotile after activation. The post-thawing motility was higher for sperm frozen with a non-activating diluent containing 2.84 M DMSO in saline (117 mM NaCl) than in an activating glycerol (2 M) medium in dilute sea water (300 mOsm). Post-thawing motility was higher for a dilution rate of 1:5 (semen:diluent) than 1:2 or 1:11 but was similar when frozen semen was thawed at 10 degrees, 20 degrees or 30 degrees C. For semen stored at a range of volumes as pellets frozen on dry ice (0.2 to 2.0 mL) or straws frozen in liquid nitrogen vapor (0.25 to 0.5 mL) and thawed in a waterbath at 20 degrees C, the post thawing motilities were similar even though the patterns of cooling and thawing differed markedly between methods of freezing and sizes of pellets and straws. PMID- 11051330 TI - Reproductive performance of purebred landrace and Yorkshire sows in Thailand with special reference to seasonal influence and parity number. AB - The purpose of this study was to analyze reproductive performance in purebred Landrace and Yorkshire sows with special reference to seasonal influence and parity number, under tropical conditions where day length is almost constant throughout the year. Data from three purebred sow herds in Thailand during the period from 1993 to 1996 were analyzed. The two breeds were present in all three herds. The analysis comprised records of 3848 Landrace sow litters and 2033 Yorkshire sow litters. The statistical models included the fixed effects of month, year, parity, breed of the sow, herd, and two-way interactions of breed parity, breed-herd, breed-month, breed-year, parity-month, month-herd, year-herd and month-year. The random effect of sow within breed was included in all models. Analysis of covariance was performed to analyze the effect of temperature, humidity and heat index on number of total born per litter (NTB), weaning to first service interval (WSI) and farrowing rate (FR). Landrace sows had significantly higher NTB (0.6 piglets), number of live born per litter (0.5 piglets), and average birth weight (0.13 kg) than Yorkshire sows (P<0.001). Farrowing rate was 3.9% higher in Landrace sows than in Yorkshire sows (P<0.01). However, Yorkshire sows had significantly shorter WSI (P<0.001) and significantly higher proportion of sows served within 7 days after weaning (P<0.01) than Landrace sows. No breed differences were found in number of stillborn per litter and weaning to conception interval. Parity had significant effect on all reproductive parameters analyzed. Number of total born and live born per litter was significantly lower for sows farrowing during the rainy season than in other seasons. Farrowing rate was low for sows mated during the hot and rainy season. Weaning to service interval and WSI7 were prolonged for sows weaned during the hot and rainy season. Reproductive performance was significantly unfavorably influenced by elevated temperature and heat index after mating (NTB and FR) or during lactation (WSI). PMID- 11051331 TI - Use of cloprostenol as an abortifacient in the llama (Lama glama). AB - As part of a larger project investigating the development and heritability of choanal atresia glama), it was necessary to develop a protocol for aborting llamas at various stages of gestation. Twenty-seven animals between 4 and 7 mo of gestation were successfully aborted a total of 53 times following two 250 microg intramuscular injections of cloprostenol at 24 h intervals. Abortion was induced once in 10 animals and multiple times (range 2 to 5) in 17 animals. Twenty-four animals (45.2%) aborted 3 d following the first injection, with 20 animals (37.7%) aborting 4 d post prostaglandin administration. Other animals aborted at 2 d (n=6, 11.3%), 5 d (n=2, 3.8%), and 7 d (n=1, 1.9%) following drug administration. Forty-nine (92.5%) of the abortions occurred following a single series of injections, while 4 animals (7.5%) aborted following a second series of injections. No confirmed pregnant animals failed to abort following the second series of cloprostenol injections. Conception rates in animals rebred 2 to 4 wk following an abortion were comparable to those of untreated animals in the research herd. Unlike the severe hypertension and death that has been reported following dinoprost tromethamine administration in the llama, no adverse reactions were observed in this study following cloprostenol administration. The results demonstrate that llamas can be safely and effectively aborted up to 7 mo of gestation (normal full term gestation = 342 +/- 10 days) without adverse effects on subsequent fertility. PMID- 11051332 TI - Topographical clinical esophageal manometry: a better mousetrap or manometric overkill? PMID- 11051333 TI - Rome? Manning? Who cares? PMID- 11051334 TI - Completing colonoscopy. PMID- 11051335 TI - Portal hypertensive gastropathy: much ado about nothing? PMID- 11051336 TI - Latin-American Consensus Conference on Helicobacter pylori infection. Latin American National Gastroenterological Societies affiliated with the Inter American Association of Gastroenterology (AIGE). PMID- 11051337 TI - The role of diet and lifestyle measures in the pathogenesis and treatment of gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) and the complications associated with it are very common in the Western world. There has been speculation for many years that certain dietary and lifestyle factors may play a role in the pathogenesis or course of GERD. However, a wide-ranging review of the available data reveals conflicting findings regarding the impact of most of these factors. In addition, the majority of the studies concerned have been based on small numbers of patients, and in most cases these studies do not meet the criteria for evidence based medicine. Consequently, any advice given on modifying diet and/or lifestyle in the management of GERD represents a form of empirical therapy. A general consensus on the control of GERD through alterations in diet and lifestyle factors could hardly be based on the results of clinical or outcome studies. In practice, however, the treatment of GERD is oriented toward the individual patient's symptoms, and includes offering reasonable advice on how to adapt to personal dietary intolerance and lifestyle factors. PMID- 11051338 TI - Serotonin: a mediator of the brain-gut connection. PMID- 11051339 TI - Colorectal cancer complicating ulcerative colitis: a review. PMID- 11051340 TI - Application of topographical methods to clinical esophageal manometry. AB - OBJECTIVE: Topographical manometric methods have improved the understanding of esophageal peristalsis in research applications but require a large number of recording sensors. Commonly used methods limited to four sensors were compared to topographical methods to determine whether the latter also had significant clinical utility. METHODS: Two hundred twelve patients referred for esophageal manometry were studied with a data acquisition system having 21 intraluminal recording sites, and the findings were analyzed independently using both limited (pull-through plus four recording sites) and topographical approaches (all sites). Discrepant results were clarified using supportive clinical data. RESULTS: The two methods were in diagnostic agreement in 187 cases (88.2%). Topographical methods correctly identified all 26 patients with achalasia within the group with aperistalsis (n = 36). The limited methods could not confidently identify six achalasia patients and were significantly less effective in segregating aperistaltic disorders (p < 0.05 across methods). Topographical methods alone detected evidence of incomplete lower esophageal sphincter relaxation in 12 additional patients, eight of whom had clinical data supporting the findings. Topographical methods identified the upper margin of the lower sphincter in all but three subjects (1.4%); limited methods could not identify this location in these and five additional subjects (3.8%) and differed from the topographical measurement by > or = 2 cm in 11.9% of cases. CONCLUSIONS: Topographical methods are more accurate than commonly used methods in diagnosing the type of severe motor dysfunction and provide additional information important in the clinical practice of esophageal manometry. PMID- 11051341 TI - Effect of different recumbent positions on postprandial gastroesophageal reflux in normal subjects. AB - OBJECTIVE: Gastroesophageal reflux (GER) is increased in the right compared to the left recumbent position. Esophageal acid exposure is related to the acidity at the cardia, but the effect of body position on the acidity at the cardia has not yet been investigated. We aimed to investigate the mechanisms underlying increased esophageal acid exposure in the right recumbent position. METHODS: On 2 separate days a 4-h combined esophageal and lower esophageal sphincter (LES) manometry and pH recording of esophagus, gastric cardia, and corpus was performed in the right and left recumbent position after a high fat meal in 10 healthy subjects. RESULTS: In the right recumbent position a prolonged esophageal acid exposure (7.0% vs 2.0%, p < 0.03), a higher incidence of reflux episodes (3.8 vs 0.9/h, p < 0.03), more transient LES relaxations (TLESRs) (6.5 vs 3.2/h, p < 0.03), and higher percentage TLESRs associated with reflux (57.0% vs 22.4% p < 0.03) was recorded than in the left supine position. Acidity at gastric cardia and corpus was not affected by body position. CONCLUSIONS: Increased esophageal acid exposure in the right recumbent position relative to the left recumbent position is the result of a higher incidence of GER episodes caused by an increased incidence of TLESRs and higher percentage of TLESRs associated with GER. Body position does not affect the acidity at the gastric cardia and corpus. PMID- 11051342 TI - A cost-minimization analysis of alternative treatment strategies for achalasia. AB - OBJECTIVE: The aim of this study is to compare the costs per cure of alternative strategies for the treatment of achalasia. METHODS: A cost-minimization model compared three strategies for otherwise healthy adults of any age with achalasia: 1) laparoscopic Heller myotomy with fundoplication (LHM); 2) pneumatic dilation (PD), with LHM reserved for treatment failures; 3) botulinum toxin (Botox) injection of the lower esophageal sphincter, with PD reserved for treatment failures. Probabilities of short- and long-term efficacy, treatment failure, symptomatic recurrence rates, and complications were derived from the published literature. Only direct costs were considered during the 5-yr time horizon. RESULTS: Respective reference case costs per cure of PD, Botox, and LHM strategies were $3,111, $3,723, and $10,792. Despite short- and long-term efficacy of 96% and 94%, respectively, the LHM strategy was most costly. Initial PD remained less costly than initial Botox, provided that rates of PD efficacy and perforation were > or = 70% and < 9.5%, respectively, and cost of a Botox session was > or = $450. The results were not sensitive to the probabilities of short- and long-term response to Botox, recurrence after PD, LHM efficacy, and post-LHM gastroesophageal reflux disease, nor to the costs of LHM and PD. CONCLUSIONS: For otherwise healthy patients with achalasia, initial PD is the least costly strategy provided that the PD perforation rate remains < 10%. Initial Botox is less costly only when nonendoscopic-related costs decrease by 25%. Initial LHM is the most costly strategy under all clinically plausible scenarios. Subsequent analyses should include a longer time horizon and an assessment of patient ference for each strategy. PMID- 11051343 TI - Effect of cholecystectomy on gastroesophageal and duodenogastric reflux. AB - OBJECTIVE: The majority of patients experience resolution of their symptoms after cholecystectomy, but a minority either find their symptoms unchanged or complain of new upper GI symptoms. It has been suggested that the effect of cholecystectomy on upper GI motility, sphincter function, or bile delivery may account for these postoperative symptoms. We aimed to determine whether cholecystectomy affects gastroesophageal reflux or duodenogastric reflux by using 24-h ambulatory pH and gastric bilirubin monitoring before and after surgery. METHODS: Seventeen symptomatic patients with gallstones underwent 24-h ambulatory esophageal and gastric pH-metry and gastric bilirubin monitoring. Helicobacter pylori status was ascertained in all patients by 14C urea breath test and serology. Combined pH and bilirubin monitoring was repeated 3 months after cholecystectomy. Eleven healthy subjects served as a control group. RESULTS: Three (17%) patients complained of persistent or new symptoms after surgery, whereas 14 (83%) patients were asymptomatic. Two patients (12%) underwent open cholecystectomy, and (88%) had the operation performed laparoscopically. No significant differences were detected in esophageal acid exposure (pH < 4), gastric alkaline shift (pH > 4), or gastric bilirubin exposure (absorbance > 0.14) after surgery. Three (17%) patients tested positive for Helicobacter pylori; the presence of infection did not appear to affect pre- or postoperative values. CONCLUSIONS: Cholecystectomy does not result in increased bile reflux into the stomach or increased gastroesophageal acid reflux. Those patients who had increased postoperative duodenogastric reflux were entirely asymptomatic. The symptoms of postcholecystectomy syndrome are unlikely to be related to increased duodenogastric reflux after surgery. PMID- 11051344 TI - Toward office-based measurement of gastric emptying in symptomatic diabetics using [13C]octanoic acid breath test. AB - OBJECTIVE: Current methods for measuring gastric emptying by breath test require sampling over several hours and are too inaccurate for clinical use. The aim of this study was to develop an office-based method for measuring gastric emptying of solids in patients with diabetes using a [13C]octanoic acid breath test. METHODS: In 22 symptomatic diabetic patients (17 insulin-dependent diabetes, 5 non-insulin-dependent diabetes) and 6 controls, we simultaneously measured gastric emptying of an egg meal (420 kcal) by scintigraphy and [13C]octanoic acid breath test. Conventional (nonlinear) methods for scintigraphic and [13C]octanoic acid breath test emptying and generalized linear regression method to predict scintigraphic half-life (t(1/2)) using four breath samples obtained during the first 3 h. RESULTS: Despite 8 h of breath sampling, the t(1/2) estimate using the conventional method was markedly different from the scintigraphic value (delta t(1/2): median, 113 min; range, 19-282 min). The generalized linear model (using samples at baseline, 30, and 120 or 150 min) yielded predicted scintigraphic tLAG and t(1/2) that were more accurate than the conventional method; mean standard deviations of differences were 16 and 27 min, respectively. Breath test correctly assessed normal or prolonged emptying in 21 of 22 patients. CONCLUSIONS: The [13C]octanoic acid breath test can be simplified to measure gastric tLAG and t(1/2) and can be expected to correctly identify normal t(1/2) in symptomatic diabetics. Further refinement of the model will need to include studies of patients with markedly delayed t(1/2). PMID- 11051345 TI - Influence of Helicobacter pylori eradication therapy on 13C aminopyrine breath test: comparison among omeprazole-, lansoprazole-, or pantoprazole-containing regimens. AB - OBJECTIVE: Proton pump inhibitors and antimicrobial agents are widely used to eradicate Helicobacter pylori (H. pylori) infection. In the general population the prevalence of infection and of polypharmacy increases the possibility of drug drug interactions during H. pylori eradication therapy. The purpose of the present study was to assess the prevalence, degree, and clinical relevance of metabolic interference with the cytochrome P450 enzymatic system occurring during 1 wk of administration of omeprazole, lansoprazole, or pantoprazole followed by the association of clarithromycin and metronidazole for another week. The 13C aminopyrine breath test (ABT) was chosen to screen for possible interactions. METHODS: We studied 30 patients referred to our Unit for H. pylori eradication therapy. They were randomized to receive either omeprazole (20 mg b.i.d.), lansoprazole (30 mg b.i.d.), or pantoprazole (40 mg b.i.d.) for 2 wk. During the second week clarithromycin (250 mg b.i.d.) and metronidazole (500 mg b.i.d.) were added. ABT was performed before, and at the end of the first and second week of therapy. Percentage of the administered dose of 13C recovered per hour at the peak (percent 13C dose/h at the peak) and cumulative percentage of administered dose of 13C recovered over time at 120 min (percent 13C dose cum120) were the ABT evaluated parameters. RESULTS: At baseline all patients showed a normal liver function. In individual patients during treatment we observed various liver metabolic interactions both as inhibition and induction, as well as after the first and the second week of therapy. However, mean modifications of the ABT parameters during the 2 weeks of therapy were not statistically significant compared to baseline values. None of the patients who had ABT variations complained of side effects. CONCLUSIONS: H. pylori eradication therapy interferes with cytochrome P450-dependent liver metabolic activity. However, the clinical relevance of these metabolic interactions is not yet apparent, and further investigation is needed. H. pylori eradication therapy appears safe, but these interactions should be considered in the choice of proton pump inhibitor and antimicrobial agents. PMID- 11051346 TI - Implication of NF-kappaB in Helicobacter pylori-associated gastritis. AB - OBJECTIVE: Transcription factor NF-kappaB plays a pivotal role in inflammatory responses by up-regulating mRNA expression of bioactive molecules such as chemokines and adhesion molecules. The present study was designed to elucidate the implication of NF-kappaB in Helicobacter pylori-associated gastritis (HAG). METHODS: We examined 41 patients with HAG and 18 H. pylori-negative control subjects. Expression of activated NF-kappaB was studied in situ by immunohistochemistry using alpha-p65 mouse monoclonal antibody (alpha-p65 mAb), which recognizes activated NF-kappaB. To identify the cell types in which NF kappaB was activated, we performed immunohistochemical analysis using antibodies against vascular endothelial cells, macrophages, and B and T lymphocytes. We also examined the colocalization of activated NF-kappaB with the of intercellular expression adhesion molecule-1 (ICAM-1) on endothelial cells. We measured the levels of NF-kappaB-dependent chemokines including interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1), regulated on activation normal T-cell expressed and secreted (RANTES) and macrophage inflammatory protein-1alpha (MIP 1alpha) in antral mucosa by ELISA (ELISA). RESULTS: Activated NF-kappaB was detected in the nuclei of epithelial cells in antral mucosa, especially of patients with HAG. NF-kappaB positivity index (NF-kappaB PI), representing the percentages of epithelial cells with positive nuclear staining for activated NF kappaB, was significantly higher in patients with HAG than in H. pylori-negative controls. NF-kappaB PI correlated significantly with histological scores of gastritis. Moreover, activated NF-kappaB was identified in the nuclei of vascular endothelial cells, macrophages, and B lymphocytes within the lamina propria in HAG. Colocalization of activated NF-kappaB with ICAM-1 expression in the same endothelial cells was demonstrated. The IL-8 levels significantly correlated with the NF-kappaB PI. CONCLUSIONS: In addition to epithelial cells, macrophages, vascular endothelial cells, and B lymphocytes contained activated NF-kappaB. In these cells, activated NF-kappaB may be involved in the inflammation process in HAG through the up-regulation of chemokines or adhesion molecules. PMID- 11051347 TI - Which dyspepsia patients will benefit from omeprazole treatment? Analysis of a Danish multicenter trial. AB - OBJECTIVE: The effect of omeprazole therapy in dyspepsia is unpredictable. The aim of this study was to identify patient characteristics and symptoms associated with the omeprazole response to improve selection of patients for empirical treatment with omeprazole. METHODS: Data from a randomized controlled trial of 471 patients with ulcer-like or reflux-like dyspepsia treated with omeprazole 20 mg daily (243 patients) or placebo (228 patients) for 2 wk were studied using logistic regression analysis. The patients were randomly divided into a model sample (N = 236) for modeling the association between the omeprazole response and descriptive variables, and a test sample (N = 235) for testing the obtained model. RESULTS: In the model sample a high body mass index, the use of antacids or H2-blockers within the last month, or pain at night time were independently associated with a good omeprazole response, whereas the presence of nausea was associated with a poor omeprazole response. Using these variables combined into a therapeutic index, the independent test sample patients could be classified into predicted good (N = 56), medium (N = 88), and poor omeprazole responders (N = 91). In these groups the observed therapeutic gain of omeprazole (omeprazole response minus placebo response) was 39.4%, 19.3%, and 4.6%, respectively (p = 0.013). For clinical use, an easy-to-use pocket chart to obtain the therapeutic index in a given patient has been devised. CONCLUSIONS: In dyspepsia the identification of potential responders to omeprazole can be improved by considering certain patient characteristics and symptoms associated with the omeprazole response. Applying these data using a simple pocket chart may assist decision about empirical omeprazole therapy in patients with dyspepsia in general practice. PMID- 11051348 TI - Factors that predict incomplete colonoscopy: thinner is not always better. AB - OBJECTIVE: The aim of this study was to determine whether anatomic factors such as body mass index (BMI) impacts the success rate of cecal intubation during colonoscopy. METHODS: We retrospectively reviewed the cecal intubation rate of 2000 colonoscopies performed at our institution from March 1997 to March 1999. The analysis sample was composed of charts for all incomplete procedures and a sample (23%) of complete examinations that were randomly selected. Data collected included age, gender, height, weight, bowel habits, abdominal surgery, psychiatric medication use, the presence of diverticular disease, amount of sedation administered, and location and reason for halting the examination. Patients were divided by BMI: thin (BMI < or = 22.1), average weight (BMI > 22.1 25.0), overweight (BMI = 25.1-29.9), and obese (BMI > 30). RESULTS: Colonoscopies in women had a lower adjusted completion rate (94.8%) than in men (98.2%) (p < 0.005). A low BMI in women was predictive of an incomplete examination (p < 0.001). Factors that did not predict incomplete examinations in women included age and previous hysterectomy. The small number of male patients with an incomplete examination (n = 16) precluded accurate identification of any factors. CONCLUSIONS: Women with a low BMI (especially < 22) were more likely to have an incomplete procedure. This finding may have implications for colorectal cancer screening in female patients. PMID- 11051349 TI - Clinical analysis of autoimmune-related pancreatitis. AB - OBJECTIVE: Several investigators have reported on autoimmune-related pancreatitis, but the clinical findings and pathophysiology still remain unclear. To clarify it, we analyzed eight patients with autoimmune pancreatitis. METHODS: We evaluated clinical findings in eight patients (four men and four women) with autoimmune-related pancreatitis. Patients were aged 45-73 yr (mean, 57.5 yr). We examined blood chemistry and immunological studies, including autoantibodies against lactoferrin or carbonic anhydrase II, and compared ERCP images with clinical findings. In two patients, we studied the subset of lymphocytes infiltrating in the pancreas by immunohistochemistry and flow cytometry. RESULTS: Four of eight patients had jaundice, two had renal dysfunction, two had abdominal pain, and two had back pain. Three patients were complicated with other autoimmune diseases. Three patients showed abnormal pancreatic exocrine function by an N-benzoyl-L-tyrosyl-para-aminobenzoic acid excretion test. Antinuclear antibody was detected in four of eight patients, antilactoferrin antibody in three of six, anticarbonic anhydrase II antibody in two of six, antismooth muscle antibody in two of seven, and rheumatoid factor in one of eight. All eight patients showed segmental stenosis of the main pancreatic duct by ERCP. Four patients showed stenosis of the common bile duct as well as the pancreatic duct. Microscopic findings showed infiltration of CD4-positive lymphocytes around the pancreatic duct, and HLA-DR was expressed on both CD4-positive cells and pancreatic duct cells. In two patients, stenosis of the pancreatic duct improved by prednisolone. CONCLUSIONS: Autoimmune mechanism may be involved in some patients with idiopathic pancreatitis associated with hypergammaglobulinemia. PMID- 11051350 TI - Association of diabetic ketoacidosis and acute pancreatitis: observations in 100 consecutive episodes of DKA. AB - OBJECTIVE: The aim of this study was to evaluate the incidence, pathogenesis, and prognosis of acute pancreatitis (AP) in diabetic ketoacidosis (DKA). DKA is associated with nonspecific increase in serum amylase levels. Autopsy studies, on the other hand, had previously raised the issue of pancreatic necrosis in patients with DKA. However, the incidence, pathogenesis and prognosis of AP in the setting of DKA has not been prospectively evaluated. METHODS: This is a prospective evaluation of 100 consecutive episodes of DKA during a period of 13 months starting in January 1998, in a university hospital in New York City. In addition to careful history, complete blood count, arterial blood gas estimation, and a comprehensive metabolic assay, serum amylase, lipase, and triglyceride levels were estimated on admission and 48 h later. All patients with abdominal pain or elevated serum levels of amylase or lipase (more than three times normal) or triglyceride levels >5.65 mmo/L (500 mg/dl) had a CT scan of the abdomen. The diagnosis of AP was confirmed when pancreatic enlargement or necrosis on contrast enhanced CT scan was seen. RESULTS: Eleven patients (11%) had AP. History of abdominal pain, not a feature on admission to include AP in the differential diagnosis, was elicited subsequently in eight patients. Abdominal pain was absent in two and one was comatose on admission. The etiology of AP was hypertriglyceridemia in four, alcohol in two, drug induced in one, and idiopathic in four patients. The hypertriglyceridemia was transient in four patients and resolved once the episode of DKA was corrected. Lipase elevation was noted in 29% and amylase elevation in 21% of all patients with DKA. Similar to increased amylase levels, serum lipase levels were also noted to be high in the absence of CT evidence of AP. CONCLUSIONS: DKA may mask coexisting AP, which occurs in at least 10-15% of cases. The pathogenesis of AP in DKA varies, but at least in some transient and profound hyperlipidemia is an identifiable factor. AP is more likely to be associated with a severe episode of DKA with marked acidosis and hyperglycemia. Ranson's prognostic criteria are not applicable to assess the severity of AP in DKA because they overestimate the severity. Severity index based on CT findings appears to better correlate with outcome. Elevation of serum lipase and amylase occur in DKA, and elevation of lipase levels appears to be less specific than amylase levels for the diagnosis of AP in the diagnosis of DKA. Although in this study AP in DKA appeared to be mild, a definite conclusion with regard to the severity should be based only on a much larger number of patients, as only 20% of patients with AP in general have serious disease. PMID- 11051351 TI - Clinically significant gastrointestinal bleeding in critically ill patients in an era of prophylaxis. AB - OBJECTIVE: Clinical studies examining stress-related gastrointestinal bleeding in critically ill patients vary in their clinical definitions and assessment of clinical significance. Although there is evidence that routine prophylaxis decreases stress-related gastrointestinal bleeding, recent studies indicate a decreasing incidence, independent of the use of prophylactic medications. The purpose of this study was to determine the incidence of and risk factors for clinically significant, endoscopically proven gastrointestinal bleeding in critically ill patients. METHODS: A database (prospectively collected data) of 8338 patients admitted to the surgical and medical intensive care units at major tertiary care center from July 1988 to April 1995 was examined. All patients with significant upper gastrointestinal bleeding as defined by a drop in hemoglobin of >20 g/L and endoscopic evidence of an upper GI tract source were identified. Risk factors for GI bleeding from stress ulceration were compared in bleeding and nonbleeding patients. A case-control study analyzing risk factors for bleeding in the abdominal aortic aneurysm subgroup was performed. RESULTS: After exclusion criteria, 12/7231 (0.17%) patients had clinically significant, endoscopically proven bleeding. Significant risk factors included age, septic shock, abdominal aortic aneurysm repair, and nutritional support. Intensive care unit stay was prolonged in patients with stress-related bleeding. There was no difference in incidence of hypotension, clamp time, APACHE score, or operating room time in patients with abdominal aortic aneurysm repair as compared with controls. CONCLUSIONS: In an intensive care unit where stress prophylaxis is widely used, clinically important gastrointestinal bleeding is uncommon. Further study is needed to define the optimal prophylaxis regimen and the role for its selective use in high-risk patients. PMID- 11051352 TI - Provocative angiography in patients with gastrointestinal hemorrhage of obscure origin. AB - OBJECTIVE: A standard diagnostic evaluation including upper and/or lower endoscopy, tagged red blood cell scintigraphy, and visceral angiography identifies the source of GI bleeding in the majority of patients who present with acute GI hemorrhage. However, in a small group of patients the source of bleeding remains obscure; this form of GI hemorrhage is uncommon but represents a considerable diagnostic challenge. Some investigators have advocated provocation of bleeding with vasodilators, anticoagulants, and/or thrombolytics in association with tagged red blood cell scans or angiography. Unfortunately, the available literature on this topic is limited. Therefore, the purpose of this study is to report our experience with provocative GI bleeding studies. METHODS: The radiology databases at Duke University Medical Center and the Durham Veterans Administration Medical Center were reviewed from 1994 to 1999. Any patient who received a vasodilator, anticoagulant, or thrombolytic to induce bleeding during a tagged red blood cell scan or visceral angiogram was included. RESULTS: Seven provocative bleeding studies were performed on seven patients. All patients underwent a visceral angiogram with intra-arterial administration of tolazoline (a vasodilator), heparin (an anticoagulant), and/or urokinase (a thrombolytic). Of the seven provocative studies, only two induced angiographically identifiable bleeding. Both of these patients underwent surgical therapy. There were no complications attributed to the provocative bleeding studies. CONCLUSIONS: These results suggest that provocative GI bleeding studies can be performed safely. However, because an active bleeding source was identified in only a small proportion of patients, we believe that further study is required to optimize patient selection and to clarify the cost-effectiveness of this approach in patients with GI hemorrhage of obscure origin. PMID- 11051353 TI - Esophageal dilation for endosonographic evaluation of malignant esophageal strictures is safe and effective. AB - OBJECTIVE: Endoscopic ultrasound (EUS) is accepted as the most accurate modality for T- and N-staging of esophageal cancer, but some malignant strictures prevent passage of the echoendoscope beyond the level of the tumor. This incomplete evaluation may decrease staging accuracy. Previous studies have yielded conflicting results regarding the safety and efficacy of esophageal dilation for EUS. METHODS: We prospectively evaluated 267 consecutive patients undergoing EUS for esophageal carcinoma staging at our institution over a 66-month period to determine the number of patients requiring dilation for EUS examination, the success of dilation, safety of dilation, and clinical importance. RESULTS: Among 267 endosonographic examinations of the esophagus, 81 (30.3%) required dilation to advance the echoendoscope beyond the level of the stricture. After dilation was performed, the echoendoscope could be passed through the stricture in 69 patients (85.2%), and in 63 of 67 of the patients dilated to > or = 14 mm (94.0%). No complications have occurred secondary to the dilations performed to permit completion of the endosonographic examination. Tumor staging by EUS after dilation was T2 (14.8%), T3 (56.8%), and T4 (21.0%), nodal staging N0 (14.6%) and N1 (75.3%); and M1 (9.9%). CONCLUSIONS: We conclude that incremental, stepwise dilation of malignant strictures to 14 mm is safe and effective in permitting echoendoscope passage beyond the stenosis. The presence of a malignant stricture does not seem to diminish the utility of EUS staging of esophageal cancer. PMID- 11051354 TI - A comparison of the Rome and Manning criteria for case identification in epidemiological investigations of irritable bowel syndrome. AB - OBJECTIVE: The aim of this study was to estimate the prevalence of irritable bowel syndrome using different standard definitions (Rome and Manning criteria) and to determine the degree of agreement between these definitions. METHODS: A population-based, cross-sectional survey study was conducted by mailing a valid, reliable questionnaire to an age- and gender-stratified random sample of residents of Olmsted County, MN, aged 30-69 yr. The threshold for a positive diagnosis of irritable bowel was varied from two to four of the six Manning criteria and from two to three of the five defecation disorders in the Rome criteria. Unadjusted as well as age- and gender-adjusted prevalence rates were calculated for each of the five definitions of IBS. Percent agreement and kappa statistics were calculated to assess agreement between the definitions. RESULTS: Questionnaires were returned by 643 of 892 eligible subjects (72% response rate). The age- and gender-adjusted prevalence of IBS varied from 20.4% using a threshold of two symptoms in the Manning criteria to 8.5% using a threshold of three defecation disorders in the Rome criteria. The percent agreement for each comparison of Manning and Rome definitions was always >90%. The kappa values ranged from 0.55 to 0.78, with the best agreement occurring between a threshold of three symptoms of Manning and two defecation disorders in Rome. CONCLUSIONS: The prevalence of IBS varied substantially depending on the specific definition of IBS used. The range of prevalence estimates in Olmsted County was similar to other published figures when IBS definition was accounted for. These findings are useful in interpreting epidemiological and clinical studies of IBS. PMID- 11051355 TI - Homocysteine in inflammatory bowel disease: a risk factor for thromboembolic complications? AB - OBJECTIVE: Patients with inflammatory bowel disease (IBD) are at increased risk for thromboembolic events. Hyperhomocysteinemia, which is an established risk factor for arterial as well as venous thrombosis, may be more prevalent in IBD because of vitamin deficiencies. METHODS: In this retrospective study, we studied the concentrations of total homocysteine (tHcy), cobalamin, folate, and pyridoxine in 231 consecutive patients with IBD, of whom 16 patients had a history of venous thrombosis, and nine a history of arterial thrombosis. Age- and gender-matched healthy volunteers served as controls (n = 102). RESULTS: Homocysteine concentrations in patients were higher (12.3 micromol/L [range 4.6 51.3] vs 11.1 micromol/L [range 3.9-27.6], p = 0.001) and hyperhomocysteinemia tended to be more prevalent in patients than in the controls (11.1% vs 5%, p = 0.07). Folate, cobalamin, creatinine, and pyridoxine concentrations were correlated with tHcy. Folate deficiency was infrequently encountered in IBD patients (4.3%). The tHcy concentration in patients with a history of venous or arterial thrombosis was not higher than in patients without a history of thrombosis (12.7 micromol/L [range 4.6-40.1] and 15.2 micromol/L (range 10.5 26.8) vs 12.3 micromol/L [range 10.5-26.8], not significant). Hyperhomocysteinemia was found in 18.8% of the patients with venous thrombosis, 11.1% of the patients with arterial thrombosis, and 10.5% of the patients without thrombosis (not significant). CONCLUSIONS: Hyperhomocysteinemia is a common phenomenon in IBD and correlates with serum folate, cobalamin, creatinine, and pyridoxine concentrations. No correlation between tHcy and a history of venous or arterial thromboembolic complications is found. Hyperhomocysteinemia does not seem to be a major contributory factor in the development of venous or arterial thrombosis in IBD patients. PMID- 11051356 TI - Fecal calprotectin levels predict colorectal inflammation among patients with chronic diarrhea referred for colonoscopy. AB - OBJECTIVE: Chronic diarrhea is a relatively common condition with multiple diverse etiologies. Stool testing may serve as a diagnostic aid to discriminate the presence or absence of organic pathology, such as colorectal inflammation. Calprotectin (a leukocyte-derived protein) and hemoglobin can be measured quantitatively from stool and represent candidate inflammation biomarkers. The aim of this study was to assess and compare the screening performance of fecal calprotectin and fecal hemoglobin among colonoscopy referral patients with chronic diarrhea of unknown origin or chronic colitis of unknown activity. METHODS: All subjects were identified prospectively and each submitted a single stool sample before purgation. Fecal calprotectin (PhiCal; Nycomed Pharma, Oslo, Norway) and fecal hemoglobin (HemoQuant; Mayo Medical Laboratories, Rochester, MN) assays were performed in separate laboratories by masked technicians. Colonoscopic and histological findings served as criterion standards for establishing the presence or absence of colorectal inflammation. RESULTS: Among 110 subjects who provided complete fecal assay data, 29 (26%) had and 81 (74%) did not have colorectal inflammation. Increased fecal calprotectin levels were significantly (p = 0.0001) associated with the presence of colorectal inflammation, whereas fecal hemoglobin levels were not (p = 0.61). Direct comparison of the fecal assays revealed that calprotectin was a more sensitive biomarker for colorectal inflammation at all specificity levels (p = 0.0001). CONCLUSIONS: In this study of colonoscopy referral patients, colorectal inflammation was reflected by fecal calprotectin but not by fecal hemoglobin levels. Assay of fecal calprotectin holds promise as a triage tool to identify inflammatory causes of chronic diarrhea. PMID- 11051357 TI - Whole gut transit scintigraphy in the clinical evaluation of patients with upper and lower gastrointestinal symptoms. AB - OBJECTIVE: In a single noninvasive, quantitative test, whole gut transit scintigraphy (WGTS) measures gastric emptying (GE), small bowel transit (SBT), and colonic transit (CT). The aim of this study was to investigate the clinical utility of WGTS in patients with functional gastrointestinal (GI) symptoms. METHODS: A total of 108 patients with either dyspeptic upper GI symptoms (n = 35) or constipation (n = 73) underwent WGTS. Patients consumed a meal of 99 m-Tc egg sandwich with 111-In DTPA in water. They were imaged every 30 min for 6 h to measure GE and SBT, and at 24, 48, and 72 h to measure CT. RESULTS: Of 108 studies, 104 were analyzable. In patients with upper GI symptoms, 14/35 (40%) had delayed GE of solids, 4/35 (11%) delayed SBT, and 11/35 (31%) delayed CT. Of those with constipation, 13/69 (19%) had delayed GE, 5/69 (7%) delayed SBT, and 43/69 (65%) delayed CT. WGTS changed the initial clinical diagnosis in 47/104 (45%) and altered patient management in 70/104 (67%) of the patients. CONCLUSIONS: Transit abnormalities of the upper GI tract and colon are common in patients with functional GI symptoms. Patients with upper GI symptoms frequently have delayed GE, whereas those with constipation tend to have predominantly delayed CT. In many patients with functional GI symptoms, there was evidence of a diffuse GI motility disorder. Whole gut transit scintigraphy is a simple, clinically useful test, as it frequently leads to a change in diagnosis and, therefore, patient management. PMID- 11051358 TI - Analysis of clinical course and long-term prognosis of surgical and nonsurgical patients with intestinal Behcet's disease. AB - OBJECTIVE: Much remains unknown about the pathogenesis of intestinal Behcet's disease. The majority of these patients are treated with surgical intervention, although it has been recently reported that a number of medical treatments are sometimes effective. Only few studies, however, have ever been undertaken to analyze the long-term prognosis of this disease. In this study, we analyzed the clinical course and the recurrences after initial therapy in patients with intestinal Behcet's disease. METHODS: We investigated 20 patients (surgical group, n = 8; nonsurgical group, n = 12) for whom the clinical courses were known for > or = 2 yr (2-23 yr). RESULTS: The surgical group tended to have higher rates of complications such as ocular and ileal lesions than the nonsurgical group. In the surgical group, 75% of the patients recurred (and were readmitted) within 2 yr, and 37.5% of the patients required reoperation for intestinal obstruction because of ulcer at the anastomosis. The percentage of peripheral CD8+ DR+ lymphocytes in the recurrent group (10.4% +/- 2.5%) was significantly higher than that in the nonrecurrent group (4.3% +/- 1.2%, p < 0.05). CONCLUSIONS: Our results indicate that more extensive disease involving the ileum and ocular lesions are markers of severity and progression to surgical crisis, and that patients requiring surgery suffer more frequent recurrences. Furthermore, an increased percentage of peripheral CD8+ DR+ lymphocytes may be a risk factor for disease recurrence. PMID- 11051359 TI - Psychological morbidity in women with idiopathic constipation. AB - OBJECTIVE: The aim of this study was to examine psychological morbidity in women with idiopathic constipation. METHODS: Three age-matched groups of 47 women with idiopathic constipation (excess straining or decreased bowel frequency), 28 healthy women, and 26 women with Crohn's disease completed a newly devised Perception of Female Self questionnaire, the Intimacy subscale of the Inventory of Interpersonal Problems, the Feminine Traits from the Bem Sex Role Inventory, and the General Health Questionnaire. Thirteen patients had a low bowel frequency (<2/wk), 30 had a normal bowel frequency, and four had increased bowel frequency (>3/day). Of 29 patients with a measured whole gut transit time, 19 had slow and 10 had normal transit. RESULTS: Patients with constipation had significantly increased psychological and social morbidity (anxiety, depression, and social dysfunction) (p = 0.022), increased somatization (p = 0.019), and less satisfaction in their sexual life (p = 0.001) than healthy women. Constipated women with slow transit or decreased bowel frequency did not differ significantly from those with normal transit and bowel frequency. Women with Crohn's disease did not differ significantly on any test from healthy controls. CONCLUSIONS: Women with idiopathic constipation have increased psychological morbidity, altered perception about female self, and impaired intimate relationships. These factors are not present in women with "organic" GI illness also associated with abdominal pain. PMID- 11051360 TI - Evaluation of the rice breath hydrogen test for small intestinal bacterial overgrowth. AB - OBJECTIVES: The aims of this study were 1) to document the sensitivity, specificity, and predictive values of the rice breath hydrogen test for small intestinal bacterial overgrowth; 2) to determine the possible influence of concurrent gastric bacterial overgrowth and gastroduodenal pH on the efficacy of this test; and 3) to investigate whether reliability is limited by an inability of small intestinal luminal flora to ferment rice or its product of hydrolysis, maltose. METHODS: Twenty adult subjects were investigated with microbiological culture of proximal small intestinal aspirate and a 3-g/kg rice breath hydrogen test. Gastroduodenal pH, the presence or absence of gastric bacterial overgrowth, and the in vitro capability of small intestinal luminal flora to ferment rice and maltose, its product of hydrolysis, were determined. RESULTS: Sensitivity of the rice breath hydrogen test for small intestinal bacterial overgrowth was 33% and remained low even when subjects with small intestinal overgrowth with oropharyngeal-type (38%) and colonic-type flora (20%) and those with concurrent small intestinal and gastric bacterial overgrowth (40%) were considered separately. Sensitivity remained suboptimal despite favorable gastroduodenal luminal pH and documented ability of bacterial isolates to ferment rice and maltose in vitro. Specificity of the rice breath hydrogen test for small intestinal bacterial overgrowth was 91%. Positive predictive value, negative predictive value, and predictive accuracy were 75%, 63%, and 65%, respectively. CONCLUSIONS: Clinical value of the rice breath hydrogen test for detecting small intestinal bacterial overgrowth is limited. The rice breath hydrogen test is not a suitable alternative to small intestinal intubation and culture of secretions for the detection of small intestinal bacterial overgrowth. PMID- 11051361 TI - Autonomic regulation of cardiac function during sleep in patients with irritable bowel syndrome. AB - OBJECTIVE: Several studies have provided evidence of abnormal autonomic activity in irritable bowel syndrome (IBS), suggesting that abnormal central nervous system-autonomic nervous system arousal mechanisms may be part of its pathophysiology. The goal was to investigate cardiac sympatho-vagal balance during waking and the different stages of sleep using heart rate variability analysis in IBS patients compared to healthy controls. METHODS: A total of 15 IBS patients (13 female, two male, mean age 34.9 +/- 2.1 yr) and 15 controls (13 female, two male, mean age 36.2 +/- 2.3 yr) were studied during 1 h of pre-sleep quiet waking and during seven-hours of sleep. Polysomnography was used for the determination of state of consciousness. Electrocardiography provided the beat-to beat intervals, which were then subjected to spectral analysis for determination of the percentage of energy in the low and high frequency bands, respectively. The low frequency/high frequency band ratio was also calculated. For each subject, heart rate variability analysis was performed using 15-min segments of waking, non-rapid eye movement sleep, and rapid eye movement sleep. RESULTS: The low frequency band power was significantly greater in IBS patients during waking. No group differences were found in high frequency band power during any state. The low frequency to high frequency band ratio was significantly greater in IBS patients during rapid eye movement sleep. CONCLUSIONS: IBS patients have greater sympathetic activity during waking and greater overall sympathetic dominance during rapid eye movement sleep. These results support the presence of autonomic abnormalities in patients with IBS. The possibility is discussed that sympathetic dominance during rapid eye movement sleep may play a role in sensitizing the gut to waking stimulation. PMID- 11051362 TI - Shared and unique environmental factors determine the ecology of methanogens in humans and rats. AB - OBJECTIVE: This study ascertains the relative contributions of genetics and environment in determining methane emission in humans and rats. There is considerable interest in the factors determining the microbial species that inhabit the colon. Methanogens. which are archaebacteria, are an easily detected colonic luminal bacteria because they respire methane. They are present in some but not all human colons and lower animal hindguts. Opinion varies on the nature of the factors influencing this ecology with some studies proposing the existence of host genetic influences. METHODS: Methane emission was measured in human twin pairs by gas chromatography, and structural equation modeling was used to determine the proportion of genetic and environmental determinants. The importance of the timing of environmental effects and rat strain on the trait of methane emission were ascertained by experiments with cohabiting methanogenic and nonmethanogenic rats. RESULTS: Analysis of breath samples from 274 adolescent twin pairs and their families indicated that the major influences on the trait of methane emission are the result of shared (53%, 95% confidence interval 39-61) and unique environmental (47%, 95% confidence interval 38-56) effects. No significant autosomal genetic effects were detected, but as observed in other studies, men (37%) were less likely to excrete methane in their breath than women (63%). Investigation of methane emission in rats indicated that environmental effects in this animal are most potent during the weaning period, with stable gut microbial ecology thereafter for some but not all rat strains. CONCLUSIONS: These results are consistent with shared and unique environmental factors being the main determinants of the ecology of this colonic microbe. PMID- 11051363 TI - Local challenge on oral mucosa with an alpha-gliadin related synthetic peptide in patients with celiac disease. AB - OBJECTIVE: Gluten-derived peptides (e.g., amino-acids 31-49 of alpha-gliadin) have been shown to cause changes typical of celiac disease in the gut. Gluten derived peptides have mostly been used in in vitro studies. The easiest access to the gastrointestinal system may be the mouth. In the present study we were interested to see whether a synthetic peptide corresponding to amino-acids 31-49 of alpha-gliadin could induce inflammatory changes in the oral mucosa after a local challenge in celiac disease patients. METHODS: The challenge was made by injecting the peptide solution at a concentration of 10 microg/ml submucosally into the oral mucosa of 10 celiac disease patients after a gluten-free diet (GFD) and 12 healthy control subjects. B and CD45RO+ T cells, mast cells, CD3+, CD4+, CD8+ lymphocytes, and alphabeta and gammadelta T-cell receptor-bearing (TcR alphabeta, TcR gammadelta) lymphocytes were counted and HLA DR expression was determined. The expression of CD25 and Ki-67 antigen was also examined. RESULTS: The peptide significantly increased the total number of T cells in the lamina propria of the celiac disease patients. The expression of T-cell activation marker CD25 (IL-2 receptor), but not that of cell proliferation marker Ki-67, was also significantly increased in the lamina propria after peptide challenge. Such a reaction was not observed in the controls. The numbers of CD3+ and CD4+ T cells in the lamina propria were also increased in celiac disease patients after the challenge. The count of TcR gammadelta+ cells was very small in the oral mucosa in celiac disease and showed no increase when the oral mucosa was challenged with the peptide. The expression of HLA DR staining was enhanced after the submucosal peptide challenge in celiac disease; however, the difference was not statistically significant. CONCLUSIONS: The results show that in the celiac disease patients after the peptide challenge the oral mucosal lamina propria responds with a nonproliferative increase of lymphocytes. Thus, submucosal challenge with the peptide 31-49 can be used as an aid in the diagnosis of celiac disease. However, further studies with optimized methodology, including various concentrations of the peptide, adjuvants, other peptides, etc., are warranted, especially because the oral mucosa provides the easiest access to an in vivo peptide challenge in celiac disease. PMID- 11051364 TI - The natural history of portal hypertensive gastropathy: influence of variceal eradication. AB - OBJECTIVE: The natural history and likelihood of bleeding from portal hypertensive gastropathy (PHG) present in patients with portal hypertension before endoscopic variceal obliteration may differ from that in patients who develop PHG during or after variceal eradication. METHODS: A total of 967 variceal bleeders who had achieved variceal eradication by endoscopic sclerotherapy in the recent past were prospectively studied. In all, 88 (9.1%) patients (cirrhosis in 54, noncirrhotic portal fibrosis in 18, and extrahepatic portal vein obstruction in 16) had distinct mucosal lesions. PHG alone was present in 78, PHG with gastric antral vascular ectasia (GAVE) in eight, and GAVE alone in two patients. PHG was graded as mild or severe and according to whether present before (group A) or after endoscopic intervention (group B). Patients underwent regular endoscopy at follow-up to see if the PHG was transitory (disappearing within 3 months), persistent (no change), or progressive. Bleeding from PHG lesions was defined as acute or chronic. RESULTS: Twenty-two (26%) patients had PHG before (group A) and 64 (74%) developed PHG after variceal eradication (group B). During a mean follow-up of 25.1 +/- 14.2 months, PHG lesions disappeared in group A in only two patients (9%), but in group B in 28 (44%) patients (p < 0.05). PHG lesions more often progressed in the former as compared to the latter (18% vs 9.4%, p = NS). The incidence of bleeding was higher in group A than group B (32% vs 4.7%, p < 0.02). Bleeding from PHG occurred in 10 patients (11.6%); seven of them were from group A, and all had either progressive (n = 3) or persistent (n = 4) lesions. CONCLUSIONS: PHG developing after variceal eradication is often transitory and less severe. If PHG is pre-existing, endoscopic therapy for varices could worsen the PHG, with a likelihood of bleeding. Such patients may be benefited by concomitant beta blocker therapy. PMID- 11051365 TI - Emergence of the precore mutant late in chronic hepatitis B infection correlates with the severity of liver injury and mutations in the core region. AB - OBJECTIVE: The reason that precore negative mutants (e-minus DNA) gradually become predominant in some patients during chronic hepatitis B virus infection is not clear. Theoretically, as long as both e-plus and e-minus DNA share the same epitopes in the core region, HBcAg-specific cytotoxic T lymphocytes (CTLs) cannot distinguish between the target peptides expressed by e-plus and e-minus DNA. Therefore, e-minus DNA may be accompanied by additional mutations in the core region, which may affect cytotoxic T lymphocyte recognition. To examine this possibility, the sequences of the precore and the entire core region of the hepatitis B virus genome were analyzed from paired serum samples in CH-B patients who experienced HBeAg to anti-HBe seroconversion (SC). METHODS: Patients were divided into two groups. Group A patients (n = 17) genome-converted to e-minus DNA in the precore region, which abolished HBeAg secretion within 3-4 yr after SC. Group B patients (n = 16) retained precore wild-type DNA for more than 3-4 yr after SC. To investigate the impact of the emergence of precore mutant type DNA on liver injury, alanine aminotransferase (ALT) levels were also examined. RESULTS: ALT flares were more severe among patients in group A than in group B. The average mean ALT level during the HBeAg negative phase of chronic infection was 54 +/- 38 in group A and 28 +/- 24 in group B. The average maximal ALT level during the HBeAg negative phase was 235 +/- 249 in group A and 83 +/- 106 in group B. Furthermore, all 17 patients in group A developed new core mutants during genome conversion. The average number of mutations in the core gene was 0.9 +/- 1.2 before genome conversion (e-plus DNA dominant phase) and increased to 2.8 +/- 1.3 for the 3-4 yr during genome conversion (e-minus DNA dominant phase). In contrast, only 56% (nine of 16) of patients in group B developed new core mutations after the loss of HBeAg. The average number of mutations in the core gene was 1.8 +/- 1.3 before SC (HBeAg-positive and e-plus DNA dominant phase), and decreased to 1.1 +/- 1.1 for 3-4 yr after seroconversion (anti-HBe-positive and e-plus DNA dominant phase). CONCLUSIONS: These data indicate that the emergence of a predominant precore negative genotype late in chronic hepatitis B virus infection is associated with the selection of additional mutations in the core gene, as well as with liver injury. PMID- 11051366 TI - Comparison of portal vein velocity and the hepatic venous pressure gradient in assessing the acute portal hemodynamic response to propranolol in patients with cirrhosis. AB - OBJECTIVE: The aim of this prospective study was to compare noninvasive Doppler sonography and invasive measurement of the hepatic venous pressure gradient (HVPG) to determine the acute portal hemodynamic response to propranolol in patients with liver cirrhosis. METHODS: In a blinded study design, portal vein velocity (PVV) and HVPG were simultaneously assessed in 11 cirrhotic patients for 4 h after oral ingestion of 40 mg propranolol. RESULTS: Both HVPG (17.2% +/- 4.3%, p < 0.0001) and PVV (15.6% +/- 2.1%, p < 0.0002) showed a highly significant reduction during the study period versus baseline. Based on HVPG measurements, four patients (36%) were classified as nonresponders. These patients had a significantly lower PVV reduction compared to the responders (responders: 18.8% +/- 2.0% vs nonresponders: 10.0% +/- 2.1%, p < 0.05). Nonresponders were identified by Doppler sonography with a sensitivity of 1.0, specificity of 0.86, and positive predictive value of 0.9 when a threshold of 20% PVV reduction 120 min after drug intake was applied. CONCLUSIONS: Doppler sonography is a useful tool for assessment of the acute portal hemodynamic effect of propranolol. To distinguish portal hemodynamic nonresponders from responders to propranolol, PVV measurements should be carried out 2 h after drug administration, and PVV reduction should be not <20% in propranolol responders. PMID- 11051367 TI - Prevalence and clinical significance of HFE gene mutations in patients with iron overload. AB - OBJECTIVE: The HFE gene contains two mutant alleles; C282Y and H63D. The C282Y mutation occurs in 55-100% of patients with hereditary hemochromatosis. The aim of our study was to re-examine the frequencies of the C282Y and H63D mutations in patients with mild and marked iron overload and in normal subjects. METHODS: A total of 82 patients with iron overload were included in this study and had hepatic iron index determination and/or quantitation of iron stores by phlebotomy. The control group consisted of 81 healthy blood donors. HFE mutation analysis was performed on leukocyte DNA using PCR-amplified genomic DNA. RESULTS: Of patients with iron overload, 70/82 (85%) were homozygous for C282Y versus 2/81 (2.5%) in the control population. Four patients had no HFE mutations despite significant iron overload, including a sister and brother (brother not included in the study group) with hepatic iron concentrations >500 micromoles/g dry weight. CONCLUSIONS: In all, 85% of our patients with iron overload were C282Y homozygotes, although a few had no HFE gene mutations. Pooled data and analysis of chromosomes considered to be at risk for H63D indicate that H63D is associated with iron overload. PMID- 11051368 TI - Prognostic indicators of risk for first variceal bleeding in cirrhosis: a multicenter study in 711 patients to validate and improve the North Italian Endoscopic Club (NIEC) index. AB - OBJECTIVE: The best known indicator of risk for first bleeding in patients with cirrhosis without previous bleeding is the index devised by the North Italian Endoscopic Club for the Study and Treatment of Esophageal Varices (NIEC index), which results from the combination of size of esophageal varices, severity of red wale marks, and Child-Pugh class. Its efficiency is far from optimal, and validation studies have reported sensitivities and specificities markedly lower than those reported in the original study. In the present study we analyzed the efficiency of NIEC index in a large series of cirrhotic patients with varices without previous bleeding. In addition, we tried to improve the effectiveness of the index by modifying it, and to validate the modifications in an independent group of patients. METHODS: A total of 627 patients were enrolled and followed until either a variceal bleeding or for a maximum of 2 yr. During this time, 117 experienced a first variceal RESULTS: Using Cox's regression analysis, size of varices, severity of red wale marks, and Child-Pugh score were significant and independent predictors of first bleeding, as already noted in the original report of the NIEC group. However, coefficients and standard errors were markedly different, and the importance of size of esophageal varices in the regression was much larger, whereas that of Child-Pugh score was much lower. According to these data, a revised index was developed (Rev-NIEC). Using receiver operating characteristic (ROC) curve analysis, the revised index showed a larger efficiency, and the area under the curve was significantly larger (0.80 +/- 0.02 vs 0.74 +/- 0.02; p < 0.01). In particular, the curve showed that for a specificity of 75%, the new index had a sensitivity of 72% compared to that of 55% of the NIEC index. Validation in an independent sample of 84 patients showed good agreement between predicted and observed risk for bleeding. Validation with the bootstrap technique also showed adequate stability of the results. CONCLUSIONS: The revised index seems to be superior to the traditional index, and may turn out to be more useful in the selection of patients for different therapeutic procedures and in the stratification of patients in clinical trials. PMID- 11051369 TI - Treatment of small hepatocellular carcinoma with percutaneous ethanol injection: a validated prognostic model. AB - OBJECTIVE: Percutaneous ethanol injection may prolong the survival of patients with small hepatocellular carcinoma associated with cirrhosis. The aim was to identify prognostic factors of survival and of local recurrence, as well as separate new lesions. METHODS: We performed Cox regression analysis in 115 consecutive patients with hepatocellular carcinoma (81 Child-Pugh class A, 34 Child-Pugh class B) treated by percutaneous ethanol injection. The validity of the model was tested by comparing predicted and observed survival in 105 independent patients from an external series. RESULTS: Overall survival rates were 89%, 63%, and 43% at 1, 2, and 3 yr, respectively. The 1-, 2-, and 3-yr survival rates were 96%, 78%, and 63%, respectively, for Child-Pugh class A patients and were 73%, 35%, 12%, respectively, for Child-Pugh class B. The albumin level was the only independent variable significantly associated with survival (p < 0.0001). The 3-yr rate of appearance of separate new lesions and local recurrence were 41% and 23%, respectively. The survival predicted by the model agreed with that observed in the independent patients. CONCLUSIONS: Survival of patients with hepatocellular carcinoma treated by percutaneous ethanol injection is related to baseline albumin level. The high rate of recurrence (both local and distant) points out the palliative role of this therapy. PMID- 11051370 TI - Treatment of chronic hepatitis C in patients who failed interferon monotherapy: effects of higher doses of interferon and ribavirin combination therapy. The Virginia Cooperative Hepatitis Treatment Group. AB - OBJECTIVE: The present study was designed to evaluate the effectiveness of interferon-ribavirin combination therapy for treatment of chronic hepatitis C virus (HCV) in patients who failed previous treatment with interferon monotherapy. METHODS: A total of 140 patients with well-documented chronic HCV who failed to achieve a virological (if HCV-RNA was assessed) or biochemical response (if HCV-RNA was not assessed) to interferon monotherapy, 3 mU three times weekly (TIW) for 3-18 months, were randomly assigned to one of three treatment groups. Group A patients were treated with 5 mU interferon TIW for 6 months. Ribavirin (1000-1200 mg daily) was added in those patients HCV-RNA positive at month 3. Group B patients were treated with 3 mU interferon TIW plus ribavirin (1000-1200 mg daily) for 6 months. The dose of interferon was increased to 5 mU TIW in those patients HCV-RNA positive at month 3. Group C patients were treated with 5 mU interferon TIW plus ribavirin (1000-1200 mg daily) for 6 months. Serum ALT and HCV-RNA were monitored during and after treatment for a total of 15 months. RESULTS: Seventeen percent of patients in group A became HCV RNA negative by treatment month 3. Adding ribavirin resulted in one additional patient becoming HCV-RNA negative. However, none of the patients in this group achieved sustained virological response. Twenty-six percent of patients in group B became HCV-RNA negative by treatment month 3. Increasing the dose of interferon from 3 to 5 mU TIW increased virological response to 30%. However, sustained virological response was observed in only 14%. Thirty percent of patients in group C became HCV-RNA negative, but sustained virological response was observed in only 12%. Sustained virological response was found to be significantly greater in patients with a nontype I HCV genotype (p < 0.002) and in patients who had a decline in HCV-RNA titer to a value < 100,000 copies/ml during their previous course of interferon monotherapy (p < 0.0001). None of the 12 sustained responders were African Americans (p < 0.013). CONCLUSIONS: Retreatment of nonresponders with interferon-ribavirin combination therapy results in limited benefit; only 13% of patients achieved sustained virological response. Response was extremely poor in African Americans and those with HCV genotype 1. PMID- 11051371 TI - Prevalence of peripheral blood cytopenias (hypersplenism) in patients with nonalcoholic chronic liver disease. AB - OBJECTIVE: Thrombocytopenia or leukopenia in patients with chronic liver disease is often attributed to functional overactivity of the spleen (hypersplenism). Despite being a fairly common phenomenon, there is a paucity of reports on the prevalence of this syndrome in stable chronic liver disease patients with or without severe fibrosis/cirrhosis. The aim of this study was to establish the prevalence of peripheral blood cytopenia in patients with nonalcoholic cirrhosis/severe fibrosis versus patients with mild or no fibrosis on liver biopsy. METHODS: This is a retrospective chart review of 235 patients who underwent a liver biopsy. One hundred ninety-one patients met strict criteria for study entry; 28 different clinical and laboratory variables were collected from their charts review, and data were then analyzed using the SPSS statistical package. RESULTS: Of the cirrhotic patients, 64% were noted to have platelet counts consistently below 150,000 (lower limit of normal in our laboratory; mean, 144.6 +/- 89.4; median, 114), whereas only 5.5% of noncirrhotic patients had thrombocytopenia (mean, 252.2 +/- 103.4; median, 238). Leukopenia (WBC, <3,500) was relatively rare in the cirrhotic/fibrotic group, having a prevalence of 5% (7.59 +/- 4.3) versus 3.3% (10.62 +/- 14.2) of noncirrhotic patients. CONCLUSIONS: Of the patients with cirrhosis, 64% had thrombocytopenia (platelet count, <150,000). The likelihood ratio of finding a platelet count of <100,000 in patients with cirrhosis, as opposed to patients without cirrhosis, is almost 12. PMID- 11051372 TI - Can large cell change and high proliferative activity predict hepatocellular carcinoma in patients with hereditary hemochromatosis? AB - OBJECTIVE: Patients with hereditary hemochromatosis are at high risk of developing hepatocellular carcinoma. This study was undertaken to define whether large cell change and nucleolar organizer regions proliferative index (marker of high proliferative activity) predict hepatocellular carcinoma development in hereditary hemochromatosis. METHODS: Histological staining for large cell change and high proliferative activity were done on baseline liver biopsies of 74 patients with hereditary hemochromatosis (52 with and 22 without cirrhosis), prospectively followed-up for 83 +/- 53 months (range, 1-230 months). RESULTS: Large cell change and high proliferative activity were found only in cirrhotic patients; 16 of 52 patients (31%) had either the large cell change or high proliferative activity. Large cell change was more frequent in patients with hepatitis B surface antigen than in those positive for hepatitis C virus (57% vs 14%, p = 0.04). Hepatocellular carcinoma developed in 7 of 16 (44%) and in 6 of 36 patients (16%) of the patients positive or negative for these morphological variables. The probability of developing hepatocellular carcinoma at 7 yr of follow-up was significantly higher in patients with large cell change or high proliferative activity than in those without. The length of follow-up from baseline histology to hepatocellular carcinoma occurrence was shorter in patients with large cell change or high proliferative activity than in those without these changes (46 +/- 36 and 109 +/- 34 months, p = 0.01). A multivariate analysis indicated that in patients with cirrhosis, large cell change or high proliferative activity (considered as a single variable), and age >55 yr were the only independent variables significantly associated with the risk of developing hepatocellular carcinoma, with a risk ratio of 4.8 (confidence interval 1.2-18.2) and 4.0 (confidence interval 1.1-15.6), respectively. CONCLUSIONS: In hereditary hemochromatosis, the presence of large cell change or high proliferative activity in patients older than 55 yr with cirrhosis should be considered a strong predictor of hepatocellular carcinoma development, especially if hepatitis B virus infection coexists. PMID- 11051374 TI - Impact of KRAS and TP53 mutations on survival in patients with left- and right sided Dukes' C colon cancer. AB - OBJECTIVE: It has been suggested that KRAS and TP53 mutated tumors might influence the phenotypic behavior of left- and right-sided colon tumors. We investigated the incidence of these mutations in left- and right-sided colon tumors and their possible influence on survival in a homogeneous group of patients with Dukes' C colon cancers. METHODS: The primary tumors of 55 patients with a sporadic Dukes' C colon cancer, all treated with adjuvant chemotherapy were analyzed for the presence of KRAS and TP53 mutations. Mutation detection of the KRAS and TP53 genes was performed on paraffin-embedded tumor material, using denaturating gradient gel electrophoresis. The 5-yr survival rates of KRAS and TP53 mutated tumors were analyzed regarding right-sided tumors (defined as tumors up to the splenic flexure) and left-sided tumors (defined as tumors from the splenic flexure to the rectosigmoid peritoneal reflection). RESULTS: KRAS mutations occurred more frequently in the right colon compared to the left colon (R = 38% (10/26); L = 10% (3/29); chi2 test: p = 0.014). KRAS mutations did not influence survival in patients with right-sided colon tumors. Patients with KRAS mutation-negative tumors in the right colon, however, had a significantly worse survival than patients with left-sided KRAS mutation-negative tumors (5-yr survival; R: 34% vs L: 65%, log-rank test: p = 0.007). TP53 mutations of a possible causative nature were found in 24 tumors (44%). Neither the incidence (R = 42% (11/26); L = 45% (13/29)) nor the survival of TP53 mutated tumors differed significantly between left- and right-sided tumors. Furthermore, survival of patients with TP53 mutation-negative tumors did not differ significantly between left- and right-sided tumors. CONCLUSIONS: There seems to be no difference in survival rate between patients with KRAS mutated and KRAS negative Dukes' C colon tumors; however, KRAS mutations are more frequently found in the right colon compared to the left colon. TP53 mutations do not have predominance for either side of the colon, and there are no differences in survival in patients with left sided versus right-sided tumors. Patients with KRAS-nonmutated tumors in the right colon did have a worse survival compared to those with such tumors in the left colon. This suggests that other genetic factors may play a role in tumor genesis in this subgroup of patients. PMID- 11051373 TI - The economic impact of the diagnosis of dysplasia in Barrett's esophagus. AB - OBJECTIVE: Cost-effective strategies for identifying patients with Barrett's esophagus who are most likely to develop cancer have not been developed. Surveillance endoscopy is currently used, and we hypothesized that more frequent surveillance intervals would identify patients with "transient positive" diagnoses of dysplasia--dysplasia found on one examination but not on subsequent ones. Our aim was to explore the potential economic impact of transient positive diagnoses of dysplasia on alternative surveillance strategies over a 10-yr period. METHODS: Data were derived from a 2-yr randomized, prospective study comparing omeprazole to ranitidine in 95 patients with Barrett's esophagus. A transient positive diagnosis of dysplasia was defined as a patient who was diagnosed with dysplasia during the study period but whose 24-month biopsies revealed no dysplasia. We calculated the number of transient positive diagnoses of dysplasia and modeled the potential economic impact of a diagnosis of dysplasia over a 10-yr period. RESULTS: Thirty patients (31%) had at least one reading of dysplasia during the study period. Nineteen patients (20%) had a transient positive diagnosis of dysplasia. During the study period, no cancers were found. A surveillance strategy of every other year and every 6 months for dysplasia would result in 1072 endoscopies over a 10-yr period at a discounted cost of $1,587,184. A total of 61% of endoscopies would be because of transient positive diagnoses of dysplasia. A strategy of yearly surveillance and every 6 months for dysplasia would result in 1404 endoscopies at a discounted cost of $2,096,733, of which 28% would result from transient positive diagnoses of dysplasia. The discounted incremental costs of more frequent surveillance in this cohort of patients over 10 yr is $509,549. CONCLUSIONS: Based on current practice strategies, transient positive diagnoses of dysplasia account for 28-61% of endoscopies in Barrett's surveillance programs. This analysis suggests that the endoscopy workload and costs associated with surveillance could be substantially reduced if patients with transient positive diagnoses of dysplasia reverted to usual surveillance after two negative examinations. PMID- 11051375 TI - Chinese alcoholic patients with esophageal cancer are genetically different from alcoholics with acute pancreatitis and liver cirrhosis. AB - OBJECTIVE: It is a mystery why some alcoholic patients acquire certain organ specific complications of alcoholism, whereas other alcoholic patients acquire different ones. The aim of this study was to investigate the differences among Chinese alcoholic patients with esophageal cancer, acute pancreatitis, and liver cirrhosis by studying the genetic polymorphisms of ADH2, ADH3, ALDH2, and P4502E1. METHODS: Liver alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), and cytochrome P4502E1 (P4502E1) are polymorphic at the ADH2, ADH3, and ALDH2 loci and the 5'-flanking region of the P4502E1. Using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, we determined the polymorphism of the above-mentioned alcohol metabolizing genes in 59 alcoholics with carcinoma of the esophagus (alcoholic esophageal Ca), 87 acute alcoholic pancreatitis patients, 116 alcoholics with liver cirrhosis (alcoholic cirrhosis), 19 alcoholics with both liver cirrhosis and acute pancreatitis (alcoholic P plus C), and 241 nonalcoholic patients. RESULTS: The results showed that the allele frequency of ALDH2*2 was significantly higher in the alcoholic esophageal Ca group than in the alcoholic pancreatitis and alcoholic cirrhosis groups. The allele frequency of ADH2*1 was significantly higher in the alcoholic esophageal Ca patients than in nonalcoholic control groups. The ALDH2*2 was significantly lower in alcoholic groups (except the alcoholic esophageal Ca group) than in nonalcoholic control groups. The allele frequencies of ADH2*1 and ALDH2*2 are higher in alcoholic patients with esophageal Ca than alcoholic patients without it. The genotype distribution of P4502E1, detected by RsaI and PstI, was not different among alcoholic patients with different organ diseases. CONCLUSIONS: The allele frequency of ADH2*1 and ALDH2*1 are different among subpopulations of alcoholics, suggesting that alcoholic patients with different specific types of organ damage are genetically different. The Chinese alcoholic patients with the ADH2*1 and ALDH2*2 allele are more susceptible to esophageal Ca. PMID- 11051376 TI - The monaural stethoscope. PMID- 11051377 TI - Transjugular intrahepatic portosystemic shunt for bleeding stomal varices associated with chronic portal vein occlusion: long-term angiographic, hemodynamic, and clinical follow-up. AB - The efficacy of the transjugular intrahepatic portosystemic shunt procedure (TIPS) in controlling refractory hemorrhage from stomal varices at the mucocutaneous junction has been previously described, but the durability of this procedure for this indication is unknown. Conservative therapy may be unsuccessful in preventing recurrent hemorrhage. Some authors believe that portosystemic shunting is the treatment of choice for patients with bleeding stomal varices who are good surgical candidates, because of the low incidence of recurrent bleeding and the longest survival. We report the 39-month angiographic and hemodynamic follow-up, and the 48-month clinical follow-up of a patient with refractory hemorrhage from stomal varices and coexistent chronic portal vein occlusion successfully treated with a TIPS procedure. PMID- 11051378 TI - Idiopathic megarectum complicating pregnancy: report of a case. AB - Pregnancy often exacerbates constipation in young women with chronic constipation syndromes. The presence of the fetus presents a challenge in both the diagnosis and treatment of these syndromes. This study was conducted to report a rare case of idiopathic megarectum complicating a pregnancy. An aggressive polyethylene glycol (PEG) regimen allowed the patient to carry the child to term and to have a normal vaginal delivery. Successful proctocolectomy was performed with coloanal anastomosis 3 months postpartum. The patient has been free of constipation for 18 months without the need for cathartics or laxatives. All efforts to avoid operative intervention should be made in constipated patients during pregnancy. This principle holds true even in the setting of dilated large bowel. Idiopathic megarectum and the management of constipation in pregnancy are discussed. PMID- 11051379 TI - A case of biliary carcinoid presenting with pancreatitis and obstructive jaundice. AB - A 43-yr-old man presented to the clinic with abdominal pain, jaundice, nausea, and vomiting and weight loss over a 6-month period. Physical exam was unrevealing other than mild epigastric tenderness. A computed tomographic scan of the abdomen revealed a mass in the head of the pancreas, which was resected at laparotomy by a Whipple's procedure. The histology showed a biliary tract carcinoid tumor. The patient had normal hydroxy-indole-acetic acid (HIAA) levels throughout. There has been no evidence of disease or tumor recurrence at 3.5 yr of follow up. PMID- 11051380 TI - Zinc-induced copper deficiency in a coin swallower. AB - In humans, acquired copper deficiency anemia is rare. This report describes a 58 yr-old man with metal pica, especially coins, who presented with symptomatic anemia. His workup led to the diagnosis of zinc-induced copper deficiency. We believe that, in this man, leaching of zinc from pennies explained the clinical and laboratory findings. This case demonstrates that health care workers should consider the possibility of zinc-induced copper deficiency when confronted with patients with unexplained anemia who have ingested coins or other zinc-containing metals. PMID- 11051381 TI - Impact of acute hepatitis B virus superinfection on chronic hepatitis C virus infection. AB - Hepatitis B virus (HBV) and hepatitis C virus (HCV) dual infection is not uncommon, but the impact of acute HBV superinfection in patients with chronic HCV infection is still unknown. Two patients with well documented chronic HCV infection were hospitalized for acute hepatitis, which was serologically confirmed to be acute HBV superinfection. One patient who was seropositive for both HBV-DNA and HCV-RNA upon admission died of hepatic failure. The other became seronegative for HCV-RNA and recovered with alanine aminotransferase normalization, seroclearance of HBsAg, and antibodies to HCV. These findings confirm that acute superinfection in patients with chronic hepatitis may increase the risk for severe hepatitis, and suggest that HBV as the newcomer may suppress the pre-existing HCV. Together with the earlier observation that acute HCV superinfection suppresses pre-existing HBV, it seems that the timing or sequence of infection is a factor influencing the outcome of viral interactions. PMID- 11051382 TI - Colorectal cancer: predicting prognosis for patients and probands using immunohistochemistry. PMID- 11051383 TI - Should nitrates be used with beta blockers to prevent variceal bleeding? PMID- 11051384 TI - Another iron in the H. pylori fire? PMID- 11051385 TI - Hepatorenal syndrome in cirrhotic patients: terlipressine is a safe and efficient treatment; propranolol and digitalic treatments: precipitating and preventing factors? PMID- 11051386 TI - Antimycobacterial therapy for Crohn's disease. PMID- 11051387 TI - Baclofen therapy for intractable hiccups induced by ultraflex esophageal endoprosthesis. PMID- 11051388 TI - Helicobacter pylori infection and family history of gastroduodenal diseases in a Japanese population. PMID- 11051389 TI - Multiple diclofenac-induced adverse effects. PMID- 11051390 TI - Lamivudine for children with chronic hepatitis B. PMID- 11051391 TI - Bezafibrate in the treatment of primary biliary cirrhosis: comparison with ursodeoxycholic acid. PMID- 11051392 TI - Colorectal cancer screening challenges health professionals. PMID- 11051393 TI - Minocycline-induced hepatitis. PMID- 11051394 TI - Hyperthyroidism with interferon-ribavirin therapy for hepatitis C: a case report and proposed treatment algorithm. PMID- 11051395 TI - Single drug therapy for Helicobacter pylori infection. PMID- 11051396 TI - Chronic hepatitis C virus infection: detection of hepatocellular carcinoma by means of contrast-enhanced color Doppler liver sonography. PMID- 11051397 TI - Primary mucoepidermoid carcinoma of the esophagus misclassified as adenocarcinoma on endoscopic biopsy. PMID- 11051398 TI - Metal duodenal stent-related cholangitis and its management. PMID- 11051399 TI - Endoscopic removal of colonic neurinoma arising from the submucosa. PMID- 11051400 TI - Fecal lactoferrin assay as a cost-effective tool for intestinal inflammation. PMID- 11051401 TI - Upper gastrointestinal bleeding from gastric submucosal arterial collaterals secondary to splenic artery occlusion: treatment by splenectomy and partial gastric devascularization. PMID- 11051402 TI - Idiopathic pancreatitis associated with ulcerative colitis. PMID- 11051403 TI - Absence of Helicobacter pylori in dental plaque assessed by stool test. PMID- 11051404 TI - ERCP experience in a community-based, private-practice setting. PMID- 11051405 TI - "Idiopathic" acute pancreatitis due to biliary sludge: prevention of relapses by endoscopic biliary sphincterotomy in high-risk patients. PMID- 11051406 TI - Isolated gastrointestinal spider nevi: potential clinical significance. PMID- 11051407 TI - Colonoscopic diagnosis of asymptomatic acute appendicitis. PMID- 11051408 TI - Role of flumazenil for paradoxical reaction to midazolam during endoscopic procedures in children. PMID- 11051409 TI - Esophageal dilation in patients with previous esophageal perforation. PMID- 11051410 TI - The high prevalence of the factor V Leiden mutation in central European inflammatory bowel disease patients. PMID- 11051411 TI - Metastatic breast carcinoma masquerading as primary colon cancer. PMID- 11051412 TI - Aspergillus fumigatus pneumonia during cyclosporine treatment for ulcerative colitis. PMID- 11051413 TI - Gastric gland mucous cells circulate in peripheral blood after endoscopic biopsy of the gastric mucosa. PMID- 11051414 TI - Nonalcoholic steatohepatitis and hepatocellular carcinoma: natural history? PMID- 11051415 TI - Purification and characterization of cellulases produced by two Bacillus strains. AB - Cellulases produced by two Bacillus strains, CH43 and HR68, isolated from hot springs in Zimbabwe, were purified to homogeneity from culture supernatants. Both enzymes had molecular mass of 40 kDa and isoelectric point of 5.4. The enzymes also resembled each other in N-terminal amino acid sequence which was Ala-Gly-Thr Lys-Thr-Pro-Val-Ala-Lys-Asn-Gly-Gln, showing 100% homology with that of endoglucanases from Bacillus subtilis belonging to glycoside hydrolase family five. The cellulases were optimally active in the pH range of 5-6.5. The optimum temperature was 65 and 70 degrees C for the endoglucanase of CH43 and HR68, respectively. The CH43 enzyme was stable at 50 degrees C in a pH range of 6-10, and HR68 at pH 6-8. Both the enzymes retained complete activity for at least 24 h at 50 degrees C. The enzymes showed highest activity with beta-glucan as substrate followed by carboxymethylcellulose. Significant activity was also observed with crystalline forms of cellulose such as filter paper and Avicel, particularly for HR68 cellulase. For carboxymethycellulose, the CH43 and HR68 cellulases had a Km of 1.5 and 1.7 mg ml(-1), respectively, and Vmax of 0.93 and 1.70 mmol glucose min(-1) mg protein(-1) respectively. The activity of the enzymes was not influenced by most metal ions at 1 mM concentration, but was increased by about 38% by Co2+. The inhibition by Hg2+ and Mn2+ was higher for CH43 than for HR68 enzyme. Ag+ inhibited the CH43 activity but stimulated the HR68 activity. The CH43 cellulase was inhibited by N-bromosuccinimide and iodoacetamide while HR68 was unaffected. PMID- 11051416 TI - Microbial degradation of phenanthrene by addition of a sophorolipid mixture. AB - The influence of sophorolipids on microbial degradation of poorly soluble phenanthrene in liquid and soil suspension culture was evaluated in the work presented. Experiments were carried out in two parts. In the first part, important basic physico-chemical characteristics of the biosurfactant and the pollutant used were determined. The critical micelle concentration (CMC) and the solubilization ratio of the biosurfactant were found to be in a good range compared with synthetic surfactants. Also, a reduction to 71% of the detectable amount of phenanthrene was measured within 4 d in soil suspension without any biotic influence. In the second part, culture experiments were done with Sphingomonas yanoikuyae, the bacterium used throughout the work presented here with the aim to assess the toxicity of the sophorolipids on these bacteria and the effect of the surfactant on biodegradation. In exponential growth tests, no toxicity up to 1 g l(-1) sophorolipids could be detected, whereas in an agar plate test, slight growth hindrance was measured at a lower concentration of 250 mg l(-1). The above mentioned data were important for planning further experiments. In the following cultivations with liquid and soil suspension media, enhancements of the biodegradation with surfactant addition were measurable. Fluorescence measurements showed that this effect was not due to an increasing biomass, but to an augmentation of bioavailability of the phenanthrene through increasing the apparent dissolved pollutant. Surfactant addition had the consequence of decreasing the residual detectable pollutant concentration (after 36 h 0.5 compared with 2.3 mg l(-1) soil suspension) and increasing the maximal degradation rate (127 instead of 80 mg l(-1) soil suspension x 10 h). Therefore, the two main problems of biological soil remediation techniques, longer process time and residual pollutants, may be solved by the use of surfactants. PMID- 11051417 TI - Root hairiness: effect on fluid flow and oxygen transfer in hairy root cultures. AB - The effect of root hairiness on fluid flow and oxygen transfer in hairy root cultures was investigated using wild-type, transgenic and root-hair mutants of Arabidopsis thaliana. The root hair morphologies of the A. thaliana lines were hairless, short hairs, moderately hairy (wild-type) and excessively hairy, and these morphologies were maintained after transformation of seedlings with Agrobacterium rhizogenes. Filtration experiments were used to determine the permeability of packed beds of roots; permeability declined significantly with increasing root hairiness as well as with increasing biomass density. Hairy roots of wild-type A. thaliana grew fastest with a doubling time of 6.9 days, but the hairless roots exhibited the highest specific oxygen uptake rate. In experiments using a gradientless packed bed reactor with medium recirculation, the liquid velocity required to eliminate external mass transfer boundary layer effects increased with increasing root hairiness, reflecting the greater tendency towards liquid stagnation near the surface of roots covered with hairs. External critical oxygen tensions also increased with increasing root hairiness, ranging from 50% air saturation for hairless roots to ca. 150% air saturation for roots with excessive root hairs. These results are consistent with root hairs providing a significant additional resistance to oxygen transfer to the roots, indicating that very hairy roots are more likely than hairless roots to become oxygen limited in culture. This investigation demonstrates that root hairiness is an important biological parameter affecting the performance of root cultures and suggests that control over root hair formation, either by use of genetically modified plant lines or manipulation of culture conditions, is desirable in large scale hairy root systems. PMID- 11051418 TI - Gentle lysis of mucous producing cold-adapted bacteria by surfactant treatment combined with mechanical disruption. AB - A procedure for the enhanced lysis of mucous producing psychrotrophic gram positive bacteria for subsequent enzyme studies is described. An initial washing of bacterial cells with Tween 80 was found to improve the degree of cell disruption in subsequent sonication or grinding with glass beads, resulting in about 20-200% increase in total soluble protein content. However, in terms of lactate dehydrogenase (LDH) activity present in the lysate, pretreatment with Tween 80 was more effective in combination with grinding, especially in the highly mucous producing strain GY11. The type of surfactant used in the pretreatment procedure before grinding strongly influenced the percentage lysis of tested strains, both in terms of released soluble protein and enzyme activity. Zymograms of LDH and glutamate dehydrogenase (GDH) activity present in the lysates also very well supported the results obtained by total protein and enzyme activity measurements. PMID- 11051419 TI - Effect of pH and stirring rate on itaconate production by Aspergillus terreus. AB - The production of itaconic acid from glucose-based media by Aspergillus terreus NRRL 1960 was found to be controlled by stirring rate and pH. When the phosphorous (P) level in the production medium was reduced to less than 10 mg l( 1), the fungal mycelium exhausted its primary growth and started to excrete itaconic acid, while it continued its secondary growth at the expense of ammoniacal nitrogen. The fermentation exhibited a mixed-growth-associated product formation kinetics, the non-growth associated production term (mI) being practically zero only when the pH was left free to change from 3.4 down to 1.85. On the contrary, when the pH was kept reducing up to a constant value by automatic addition of KOH 4 mol l(-1), the itaconate yield coefficient on the initial glucose supplied (Y(I/So)) and mI and were 0.53 g g(-1) and 0.028 h(-1) at pH 2.4 and 320 rev min(-1) and 0.5 g g(-1) and 0.036 h(-1) at pH 2.8 and 400 rev min(-1), respectively. Although the differences between mI and Y(I/So) were statistically insignificant at the 95% confidence level, the net difference in the corresponding yield coefficients for itaconic acid on mycelial biomass resulted in a maximum itaconate production rate of 0.41 g l(-1) h(-1) at pH 2.8 and 400 rev min(-1), thus showing that this operating condition is no doubt optimal for the process under study. PMID- 11051420 TI - Effects of purified endo-beta-1,4-xylanases of family 10 and 11 and acetyl xylan esterases on eucalypt sulfite dissolving pulp. AB - Sulfite dissolving pulp from Eucalyptus grandis contained approximately 3.8% O acetyl-4-O-methylglucuronoxylan with a molar ratio of xylose:4-O-methylglucuronic acid:acetyl group close to 13.6:1:6.2. The effects produced by purified endo xylanases from two different glycosyl hydrolase families (family 10 and 11) as well as acetyl xylan esterases were examined and assessed on pulp in relation to their bleaching abilities. The purified endo-xylanases hydrolyzed only a limited portion (less than 30%) of the acetylglucuronoxylan present in the pulp. The enzymes of family 10 produced acetylated xylobiose and xylotriose whereas acetylated xylobiose was not observed among the products released from the pulp by the family 11 xylanases. The esterases however were not capable of deacetylating the acetylated aldouronic acids generated by the xylanases. Regardless of the different mode of action of the endo-xylanases on dissolving pulp, their effect on pulp bleaching was not related to the amount and nature of sugars generated or the glycosyl hydrolase family. No additional brightness gain was obtained when endo-xylanases were used in conjunction with acetyl xylan esterases, suggesting that the latter do not play an important role in biobleaching of eucalypt sulfite dissolving pulps. PMID- 11051421 TI - Southern blot screening for lignin peroxidase and aryl-alcohol oxidase genes in 30 fungal species. AB - Screening to detect genes encoding lignin peroxidase (LiP) and aryl-alcohol oxidase (AAO) has been carried out with 30 fungal strain using DNA probes from genes lpo of Phanerochaete chrysosporium (encoding LiP isoenzyme H8) and aao of Pleurotus eryngii. Evidence for the presence of genes closely related to lpo was found in Bjerkandera adusta, Fomes fomentarius, Ganoderma applanatum, Ganoderma australe, Lentinula degener, Peniophora gigantea, P. chrysosporium, Phanerochaete flavido-alba and Trametes tersicolor, whereas the gene aao was detected in Pleurotus species and B. adusta. The presence of both genes was only detected in B. adusta. These results suggest that different enzymatic system, formed by enzymes encoded by different genes, are responsible for lignin degradation by white-rot fungi. PMID- 11051422 TI - On a new artificial mediator accepting NADP(H) oxidoreductase from Clostridium thermoaceticum. AB - The purification and partial characterisation of an NADP(H) dependent artificial mediator accepting pyridine nucleotide oxidoreductase (AMAPOR) from the anaerobic Clostridium thermoaceticum is described. Depending on the redox potential of the artificial mediators the AMAPOR is able to regenerate NADP+ or NADPH rendering the enzyme useful for preparative work applying NADP(H) dependent oxidoreductases. At 37 degrees C crude extracts of C. thermoaceticum have an AMAPOR activity of 5-7 U mg(-1). This is 28 degrees under the optimal growth temperature of this microrganism. Out of apparently more than 10 AMAPOR active proteins in the crude cell extracts visible after electrophoresis and activity staining on the gel, two of these proteins were isolated. They seem to be two different oligomers. According to gel electrophoresis they show apparent molecular masses of about 200 and 400 kDa. These two forms showed after SDS gel electrophoresis two monomers with apparent molecular masses of 42 and 56 kDa which we call alpha and beta. The two oligomers may have the compositions alpha2beta2 and alpha4beta4. They contain Fe/S cluster and FAD. Various amounts of the FAD were lost during the purification procedure. This loss is partially reversible after addition of FAD. The AMAPOR reacts with rather different artificial mediators such as viologens, quinones e.g. 1,4-benzoquinone or anthraquinone-2,6-disulphonate, 2,6-dichloro-indophenol and clostridial rubredoxin. Two different ferredoxins from C. thermoaceticum, oxygen or lipoamide are no substrates indicating the here described AMAPOR is not a diaphorase in the usual sense. PMID- 11051423 TI - Role of esophageal pH recording in management of chronic laryngitis: an overview. AB - Chronic laryngitis typically produces symptoms of frequent throat-clearing, soreness, decreased voice quality with use, nonproductive cough, globus sensation, and odynophagia. The endoscopic laryngeal examination usually demonstrates posterior glottic edema, erythema, and increased vascularity and nodularity. There is increasing support for the hypothesis that reflux of acidic gastric contents is often responsible for the symptoms and findings of chronic laryngitis. Prospective trials of acid suppression therapy demonstrate not only efficacy in symptom reduction, but also objective improvement in measurements of voice quality and mucosal erythema. Although traditionally considered the "gold standard" for diagnosis of reflux causing laryngitis, routine esophageal pH recording may result in false negatives in up to 50% of patients. This may confound the diagnosis of chronic laryngitis and delay treatment. Conversely, a positive study during comprehensive therapy may help identify patients who need additional treatment. A single distal probe is probably insufficient for evaluation of a supraesophageal disorder. Current recommendations for double probe pH study in the evaluation of chronic laryngitis fall into 2 categories: 1) a double-probe pH study is indicated if there is ongoing moderate-to-severe laryngitis despite antireflux precautions and proton pump inhibitor treatment for at least 6 to 12 weeks; and 2) a double-probe pH study is indicated as a baseline measurement before Nissen or Toupet fundoplication. The pH study would also be indicated in patients who have symptoms after fundoplication. There is clearly much more work to be done on the technical issues of obtaining accurate objective data related to laryngeal acidification. In addition, although acid reflux appears to be causative in many cases of chronic laryngitis, further work is indicated to identify reliable testing methods that will predict treatment success. PMID- 11051424 TI - Ambulatory pH monitoring methodology. AB - Twenty-four-hour ambulatory double-probe pH monitoring is the current "gold standard" for the evaluation of gastroesophageal and extraesophageal reflux. The following article seeks to characterize some of the areas of controversy and describes the methodology of ambulatory pH monitoring used at the Center for Voice Disorders of Wake Forest University. PMID- 11051425 TI - Application of ambulatory 24-hour multiprobe pH monitoring in the presence of extraesophageal manifestations of gastroesophageal reflux. AB - The recognition of extraesophageal manifestations of gastroesophageal reflux (GER) has gained acceptance in recent years. Objective measurements of acid reflux have been aided greatly by the advent of multiprobe pH monitoring. The development of the multiprobe pH monitor has enabled researchers and clinicians to objectively measure acid exposure in the pharynx and upper esophagus. The indications for use of this diagnostic technique for a variety of extraesophageal manifestations of GER are outlined. PMID- 11051426 TI - Value of pH probe testing in pediatric patients with extraesophageal manifestations of gastroesophageal reflux disease: a retrospective review. AB - Extended pH probe testing is often performed in patients believed to have extraesophageal symptoms of gastroesophageal reflux disease (GERD), although for this indication its diagnostic value is not well established. A retrospective review of all patients who underwent pH probe testing between 1994 and 1998 was conducted to determine the outcome of antireflux therapy in the subgroup with probable extraesophageal symptoms of GERD. Sixty-eight patients underwent antireflux therapy and had adequate follow-up after pH probe testing to be included in the study. Fifty-eight patients (85%) responded to antireflux therapy (improved, 44%; cured, 41%). The positive predictive value of distal pH probe testing was greater than 90%, but the negative predictive value was less than 50%. The reproducibility of pH probe testing on different study days was poor, but pH probe testing was helpful in assessing the adequacy of antireflux therapy. The presence of gastrointestinal symptoms did not correlate with the response of extraesophageal symptoms to antireflux therapy. Thirteen patients underwent double-probe pH studies. The mean percent time the pH was less than 4 in the upper esophagus was 2.6% (range, 1% to 9.6%). Twelve of these patients were improved or cured with antireflux therapy. Distal pH probe testing is of limited benefit in predicting whether patients with extraesophageal symptoms of GERD will respond to antireflux therapy. If extraesophageal symptoms of GERD are suspected, patients should undergo an empiric trial of antireflux therapy. Distal pH probe testing should be reserved for assessing the adequacy of antireflux therapy if symptoms persist. A prospective, randomized, controlled study will aid in determining the predictive value of double-probe pH studies in pediatric patients with probable extraesophageal symptoms of GERD. PMID- 11051427 TI - Endoscopic evaluation of swallowing as an alternative to 24-hour pH monitoring for diagnosis of extraesophageal reflux. AB - Patients with symptoms of extraesophageal reflux may not be eager to undergo 24 hour pH probe monitoring for the sake of a definitive diagnosis. It has been anecdotally noted that extraesophageal reflux can be detected during an endoscopic swallowing evaluation. The purpose of this pilot study was to demonstrate that flexible endoscopic evaluation of swallowing with sensory testing (FEESST) can be implemented to identify and subsequently treat patients with extraesophageal reflux. Over a 6-month period, FEESST was prospectively performed in 20 healthy control subjects and in 20 patients with dysphagia. The dysphagic patients did not have a history of stroke or chronic neurologic disease. Attention was specifically directed toward noting the presence or absence of reflux into the laryngopharynx during the endoscopic swallowing evaluation. None of the control subjects were noted to have reflux during FEESST, but 18 of the 20 patients with dysphagia were found to have reflux during the evaluation; this difference was statistically significant (p < .001, Fisher's exact test). We conclude that FEESST is useful as a means of diagnosing extraesophageal reflux in patients with dysphagia. PMID- 11051428 TI - Multidisciplinary airway stent team: a comprehensive approach and protocol for tracheobronchial stent treatment. AB - Tracheobronchial stents are being used with increasing frequency to treat major airway obstruction from both malignant and benign processes. Traditionally, stents have been placed via rigid bronchoscopy, flexible bronchoscopy, or fluoroscopy by members of various individual disciplines. We describe a novel multidisciplinary airway stent team (MAST) protocol for tracheobronchial stent placement and endoscopic management of major airway obstruction. A patient with symptoms of airway obstruction is generally first evaluated with a computed tomography scan and a videotaped flexible bronchoscopy. These studies are reviewed by the team otolaryngologist, pulmonologist, and interventional radiologist. A treatment plan, including the type and location of stents and the need for adjuvant therapies, is formulated. Stent placement is performed in the operating room under general anesthesia. Rigid bronchoscopy, with flexible bronchoscopy and fluoroscopy as needed, allows precise stent placement and the best use of various therapeutic methods. The MAST protocol combines the skills, knowledge, and unique therapeutic options of specialists from otolaryngology, pulmonology, and interventional radiology. This approach allows optimal stent placement and the use of other endobronchial therapies, including laser ablation, balloon dilation, photodynamic therapy, cryotherapy, and brachytherapy. A protocol with representative case reports is presented, along with a review and comparison of several of our most commonly used stents. Otolaryngologists who practice bronchoesophagoscopy, by virtue of their operative skill and knowledge of airway management, are well equipped to become leaders of MASTs and are encouraged to initiate MASTs at their institutions. PMID- 11051429 TI - Pediatric laryngopharyngeal sensory testing during flexible endoscopic evaluation of swallowing: feasible and correlative. AB - Laryngopharyngeal sensory testing can predict aspiration risk in adult patients. Its feasibility and potential role in the evaluation of pediatric swallowing is undetermined. The goals of this study were to determine the feasibility of performing laryngopharyngeal sensory testing in awake pediatric patients and to assess whether the sensory testing results correlated with aspiration during a feeding assessment or correlated with a history of pneumonia. Fiberoptic endoscopic evaluation of swallowing with sensory testing was performed in 100 pediatric patients who were evaluated for feeding and swallowing disorders. The swallowing function parameters evaluated were pooled secretions, laryngeal penetration, and aspiration. The laryngopharyngeal sensory tests were performed by delivering a pressure-controlled and duration-controlled air pulse to the aryepiglottic fold through a flexible laryngoscope to induce the laryngeal adductor response (LAR). The air pulse stimulus ranged in intensity from 3 to 10 mm Hg. The patients tested ranged from 1 month to 24 years of age, with a median age of 2.7 years. Sensory testing was completed in 92% of patients. Patients who had an LAR at less than 4 mm Hg rarely if ever had episodes of laryngeal penetration or aspiration. Those with an LAR at 4 to 10 mm Hg had variable amounts of aspiration and laryngeal penetration. The LAR could not be elicited at the maximum level of intensity (10 mm Hg) in 22 patients, who demonstrated severe laryngeal penetration and/or aspiration. Elevated laryngopharyngeal sensory thresholds correlated positively with previous clinical diagnoses of recurrent pneumonia, neurologic disorders, and gastroesophageal reflux, and correlated positively with findings of pooled secretions, laryngeal penetration, and aspiration. Laryngopharyngeal sensory testing in children is feasible and correlative. PMID- 11051430 TI - Evaluating laryngotracheal stenosis in a canine model with virtual bronchoscopy. AB - We performed a prospective masked animal study to determine whether virtual bronchoscopy, a noninvasive computed tomography technique, can accurately measure upper airway stenosis. Virtual bronchoscopy creates a 3-dimensional endoscopic image from spiral computed tomography data. Laryngotracheal stenosis was endoscopically induced in 18 dogs. The excised larynges were examined by endoscopy, virtual bronchoscopy, and macrodissection. Measurements were made of the anteroposterior (A-P) diameter, the left-right (L-R) diameter, the full length of stenosis in the sagittal plane, and the length of the tightest stenotic segment. Each measurement method was performed independently. All investigators were unaware of measurements made by others. The measurements obtained through virtual bronchoscopy and actual endoscopy were compared to those made at dissection by interclass correlation coefficients (ICCs). Endoscopy was better than virtual bronchoscopy in measuring the A-P diameter (ICC = .79, p < .0001; ICC = .42, p = .01). Both were equally effective in measuring the L-R diameter (ICC = .53, p = .0062; ICC = .52, p = .0064). The endoscopes could not assess the full length of the stenosis, whereas virtual bronchoscopy measured it fairly accurately (ICC = .72, p = .0001). Virtual bronchoscopy relatively accurately measured the length of the tightest stenotic segment (ICC = .68, p = .0002), whereas endoscopy produced measurements in only 11 of 18 larynges, and the measurements were less accurate (ICC = .45, p = .0068). Virtual bronchoscopy can provide good measurements of stenotic lesions in the airway. It is more accurate than actual endoscopy in determining the length of stenosis. It may therefore be useful as an adjunct imaging method in preoperative planning for reconstructive surgery. PMID- 11051431 TI - Age- and gender-related collagen distribution in human vocal folds. AB - The composition of the lamina propria in human vocal folds has been shown to affect vocal performance. Collagen plays a significant role in the biomechanical effects of the lamina propria. Specifically, it lends tensile strength to the rapidly oscillating fold. We obtained from a state medical examiner 38 larynges from men and women in infant, adult, and geriatric age groups. We stained the vocal folds for collagen using a picric acid stain and studied them using an image analysis system. Distributions of collagen were measured from the superficial to deep layers (from epithelium to vocalis muscle) within the lamina propria. The data showed an increase in collagen content from infant to adult stages. Infant folds had about 51% of the collagen found in all adults and in geriatric patients (p < .001). There was no significant difference between adult and geriatric folds (p < .16). There was, however, a gender difference in the amount of collagen in both adult and geriatric specimens. Female adult and geriatric folds had about 59% of the collagen found in male adult and geriatric folds (p < .001). The distribution pattern of collagen showed that most of the collagen was present in the deep layer. From these data we conclude that there are age-related and gender-related differences between male and female infant, adult, and geriatric vocal folds. Stress-strain performance studies need to be correlated with histologic findings to better study the phonetic implications of these findings. PMID- 11051432 TI - Effects of intralaryngeal carbon dioxide and acetazolamide on the laryngeal chemoreflex. AB - Sudden infant death syndrome is the leading cause of death in infants in the United States. The laryngeal chemoreflex (LCR) is thought to contribute to its pathogenesis. In adult animals, increasing levels of intralaryngeal CO2 result in a decrease in ventilatory activity. Intravenous acetazolamide (AZ) abolishes this response. The purpose of this study was to determine the effects of intralaryngeal CO2 and AZ on the LCR and respiratory physiology of piglets under normoxic and hypoxic conditions. We applied 0% or 10% CO2 in a randomized order to the larynx of 26 piglets. Intubation via tracheotomy prevented inhalation of the gas mixtures. Laryngeal stimulation was performed under normoxic conditions (PaO2 of >70 mm Hg) in 15 animals and under hypoxic conditions (PaO2 of 50 to 65 mm Hg) in 11 animals both with and without intravenous AZ (5 mg/kg). Respiratory and cardiovascular response data were recorded. Ten percent intralaryngeal CO2 has no significant effect on mean baseline respiratory rate, systemic PaCO2 or PaO2 levels, or apnea duration (p > .05). The use of AZ (versus no AZ) resulted in significantly higher baseline respiratory rates (64 versus 51 breaths per minute; p = .016), a decreased baseline systemic PaCO2 level (38.8 versus 45.9 mm Hg; p < .001), a higher baseline PaO2 level (97.9 versus 82.8 mm Hg; p < .001), shorter mean apnea durations (15.5 versus 24.8 seconds; p = .001), a higher lowest O2 saturation level after the stimulus (78.0% versus 68.4%; p = .003), and fewer profound apneas (10 of 90 versus 41 of 90 trials; p < .001). We conclude that 10% intralaryngeal CO2 does not decrease ventilatory activity in piglets and has no significant effect on the LCR. Acetazolamide, however, appears to have a protective effect against the LCR, resulting in shorter and less severe apneas. The protective effect of AZ against the LCR appears to be related to its ability to stimulate the respiratory drive and increase oxygenation at baseline. PMID- 11051433 TI - Function of a hearing aid under stressful conditions. AB - The auditory function of individuals with normal hearing was compared with that of hearing-aided subjects of similar age to determine whether amplification remediates hearing impairment under stressful auditory situations. The specific tests of listening in a competitive noise environment and identifying moderately compressed speech were introduced to adequately aided individuals. The data indicate that noise had an impact on auditory function to a much greater degree in aided individuals than in matched counterparts with normal hearing. The data derived from acceleration of simple sentences delivered to the aided group suggested that contrary to basic tonal sensitivity, the capacity to understand the stimulus was greatly compromised. The authors discuss cochlear damage and central auditory impairment as they relate to the limitations of amplification for sensorineural hearing loss. PMID- 11051434 TI - Rupture pressures of membranes in the ear. AB - The rupture pressures of the tympanic membrane, Reissner's membrane, the round window membrane, and the annular ligament have all been measured in cadaver ears from Norwegian cattle. For the tympanic membrane, a static overpressure was applied to the ear canal; for Reissner's membrane, to the endolymph; and for the round window membrane, to the perilymph. The rupture pressure of the annular ligament equals the rupture force to the footplate divided by the area of the oval window. The mean rupture pressures are 0.39 atm for the tympanic membrane, 0.047 atm for Reissner's membrane, greater than 2 atm for the round window membrane, and 29.4 atm for the annular ligament. This last pressure corresponds to 0.68 kilogram force applied to the footplate. The ruptures of the tympanic membrane appeared without exception as small tears in the pars flaccida. The rupture pressure of the tympanic membrane was also measured in a few ears from foxes. PMID- 11051435 TI - Sensitivity of the endocochlear potential level to cochlear blood flow during hypoventilation. AB - To study the relationship between endocochlear DC potential (EP) and cochlear blood flow (CoBF) under hypoxic conditions, we recorded the EP and CoBF from the basal turn of the cochlea in 21 guinea pigs. Hypoventilation for 10 minutes was induced by reducing the respiratory rate and volume. During hypoventilation, the EP declined in most of the cases to an intermediate level of the positive range in a few minutes. At the midpoint of the 10-minute hypoventilation, angiotensin II (5 microg/kg or 1 mL/kg) was infused for 60 seconds to raise the systemic blood pressure. In this experimental manipulation of systemic blood pressure, the CoBF and EP generally rose transiently. We determined the sensitivity of the EP to a CoBF change (delta) by calculating the deltaEP/deltaCoBF. More specifically, we analyzed the relationship between the deltaEP/deltaCoBF and the EPi (EP level just before angiotensin II infusion). The deltaEP was equal to the maximum EP level after angiotensin II infusion minus the EPi. The deltaCoBF was equal to the maximum CoBF value after angiotensin II infusion minus the CoBF value just before infusion. The deltaEP/deltaCoBF increased most in the range near 70% of the EPi. That is, the deltaEP/deltaCoBF was greater and the EPi was lower in the range above 70% of the EPi. To elucidate this linear correlation in the range above 70% of the EPi, we must consider several factors. In the supplementary experiments for blood gas analysis using 11 guinea pigs, most of the data of the EPi in the range above 70% were found to be obtained under conditions of a PaO2 of more than 12 mm Hg. As to the sensitivity increase of the EP to the deltaCoBF above mentioned, we propose that among several factors in the stria vascularis during hypoxemia, the activation of glycolysis in aerobic metabolism may be involved. As another possible factor, we postulate the increase in the reactive rate of the enzymatic activities that are linked with EP production and respond to the elevated cyclic adenosine monophosphate activity induced by the sympathicotonic state due to hypercapnia. PMID- 11051436 TI - Unilateral nasal nitric oxide measurement after nasal surgery. AB - This study was designed to validate and standardize a method for unilateral nasal nitric oxide (NO) measurement. Fourteen healthy volunteers and 11 patients who had undergone unilateral medial maxillectomy were enrolled. Nasal NO was measured unilaterally by means of a dual pump system, and bilateral nasal NO was measured by aspirating air through the nasal airway in series. The median unilateral NO output was 195 nL/min on the surgical side and 291 nL/min on the contralateral, surgically untreated side (p = .006). The NO output was not significantly different between nostrils in the control group (p = .82). With the bilateral technique, there was no significant difference between the surgery group and the healthy-subjects group (p = .72). The unilateral nasal NO technique is sensitive in determining unilateral differences in nasal NO production. The NO outputs from the nostrils were similar in normal subjects regardless of the nasal cycle, but were significantly lower on the operated side in the unilateral nasal surgery group. PMID- 11051437 TI - Stimulation of adenoidal lymphocytes by Alloiococcus otitidis. AB - Otitis media with effusion (OME) is characterized by persistent effusion in the middle ear cavity and by chronic inflammation in the middle ear mucosa. Alloiococcus otitidis, a gram-positive aerobic bacterium, has been isolated in middle ear effusion, and by means of sensitive polymerase chain reaction detection assays it has been detected in as many as 20% of middle ear aspirates of patients with OME. Because A otitidis may freely interact with leukocytes in the middle ear effusion, it may potentially modulate the inflammatory reaction in OME. To study the nature of these interactions, we applied an in vitro assay in which killed A otitidis bacteria were incubated with peripheral blood and adenoidal mononuclear cells. The expression of the proliferation-associated surface marker CD69 was then measured in B lymphocytes and in CD4+ helper and CD8+ cytotoxic-suppressor T lymphocytes by means of multicolor flow cytometry. Alloiococcus otitidis induced the expression of CD69 in both peripheral blood and adenoidal T and B cells. Among the T cells, the cytotoxic-suppressor T lymphocytes were preferentially activated. It was also tested whether A otitidis would have an effect in another cytotoxic and immunoregulatory system, namely, the induction of natural killer cell activity in peripheral blood mononuclear cells. However, the effect was minimal compared with that of Salmonella minnesota or Staphylococcus aureus. The results show that A otitidis has a unique immunostimulatory capacity in vitro that is mainly confined to CD8+ T lymphocytes. PMID- 11051438 TI - Vertical partial laryngectomy versus supracricoid partial laryngectomy for selected carcinomas of the true vocal cord classified as T2N0. AB - From an inception cohort of 204 patients with squamous cell carcinoma of the true vocal cord classified as T2N0 and a minimum of 3 years of follow-up, the authors compare the oncological and functional outcomes following vertical partial laryngectomy (group 1; 85 patients) and supracricoid partial laryngectomy (group 2; 119 patients). The 10-year actuarial survival estimate was 46.2% for group 1 and 66.4% for group 2. Survival was statistically more likely to be reduced (p = .019) in group 1 than in group 2. The 10-year actuarial local control estimate was 69.3% for group 1 and 94.6% for group 2. Local recurrence was statistically more likely to occur (p < .0001) in group 1 than in group 2. Salvage treatment resulted in an overall 94.1% local control rate and a 78.1% laryngeal preservation rate for group 1 and an overall 99.2% local control rate and a 94.9% laryngeal preservation rate for group 2. The 10-year actuarial nodal control estimate was 81.7% for group 1 and 93.7% for group 2. Nodal recurrence was statistically more likely to occur (p = .028) in group 1 than in group 2. The 10 year actuarial estimate for patients without distant metastasis was 84.6% for group 1 and 95.1% for group 2. Distant metastasis was statistically more likely to occur (p = .05) in group 1 than in group 2. The hospital mortality rate was 1.2% for group 1 and 0.8% for group 2. The incidence of permanent gastrostomy was 0% for group 1 and 2.4% for group 2. The incidence of permanent tracheostomy was 1.2% for group 1 and 2.4% for group 2. The incidence of completion laryngectomy due to functional problems was 1.2% for group 1 and 0.8% for group 2. PMID- 11051439 TI - Recurrent laryngeal nerve transposition in guinea pigs. AB - Improved control of prosthetic voice aids for laryngectomees might be possible to obtain with residual laryngeal motor nerve signals. We were able to recover motor signals from the recurrent laryngeal nerve (RLN) by transposing it into the ipsilateral denervated sternohyoid muscle (SH) in 8 guinea pigs. Reinnervation was monitored by electromyographic recordings from surface and intramuscular needle electrodes in awake animals. Within 4 to 14 weeks after surgery, all animals demonstrated laryngeal-like motor activity in the reinnervated SH, including activity during respiration, sniffing, swallowing, and/or vocalizing. After 3 to 6 months, the animals were reanesthetized, and nerve stimulation and section experiments confirmed the RLN as the source of reinnervation in all cases. In several animals, activity of the RLN-innervated SH was demonstrated to be correlated with that of contralateral laryngeal muscles. Histochemical analysis of the SH indicated a unilateral transformation from mostly fatigable to mostly fatigue-resistant fiber types ipsilateral to the RLN transposition, a phenotype more typical of laryngeal muscles. Thus, RLN transposition at the time of laryngectomy may be a method for salvaging laryngeal control signals that could be used to control prosthetic voice devices. PMID- 11051440 TI - Tracheal reconstruction with porous high-density polyethylene tracheal prosthesis. AB - The variety of methods used for the treatment of severe tracheal stenosis and the occasional failure to obtain lasting and consistent relief are measures of the difficulty of the problem. Presently, several surgical approaches are used for reconstructing large defects of the cervical trachea, reminding us that no single technique has gained wide clinical acceptance. We resected a U-shaped cartilage from 6 to 8 rings of the trachea of 10 New Zealand rabbits. We used a porous high density polyethylene (PHDP) prosthesis to reconstruct the cervical trachea, and did not perform a tracheotomy during this operation. We followed up the rabbits for 4 to 10 months. One of the rabbits died in the second month because of respiratory distress; another was lost in the fourth month of the study while delivering young. The others continued to thrive; in fact, one of them gave birth 3 times after our study and was well able to take care of her young. At the end of the study, the rabbits were painlessly sacrificed in order to remove the larynx and trachea and examine them histopathologically. In the histopathologic examinations, the prostheses were incorporated into the native trachea and adhered to the surrounding organs, especially to the esophagus. No mucosal irregularities were seen on the surface of the prosthesis, and all the surfaces appeared to be covered with ciliated pseudostratified epithelium. This tracheal prosthesis provides good results in rabbit tracheal reconstruction, and appears very promising for the clinical repair of tracheal defects. PMID- 11051441 TI - Pediatric temporal bone fractures. PMID- 11051442 TI - Behavior, appetite, and urinary cortisol responses by adult female pigtailed macaques to cage size, cage level, room change, and ketamine sedation. AB - Pigtailed macaques (Macaca nemestrina) and longtailed macaques (M. fascicularis) show behavioral, ecological, and possible temperament differences, and their responses to the laboratory environment might therefore be quite different. We tested pigtailed macaques under the same conditions that were investigated in a previous study with longtailed macaques, using the same comprehensive set of physiological and behavioral measures of stress. First, eight adult females' adaptation to a new room in regulation-size cages was monitored, and in the third week their responses to ketamine sedation were measured. Then they spent two weeks singly housed in each of four cage sizes (USDA regulation size, one size larger, one size smaller, and a very small cage). Half of the subjects were in upper-level cages and the remainder in lower-level cages for the entire study. Cage size, ranging from 20% to 148% of USDA regulation floor area, was not significantly related to abnormal behavior, self-grooming, manipulating the environment, eating/drinking, activity cycle, cortisol excretion, or biscuit consumption. Locomotion and frequency of behavior change were significantly reduced in the smallest cage, but did not differ in cage sizes ranging from 77% to 148% of regulation size. The only manipulation to produce an unequivocal stress response, as measured by cortisol elevation and appetite suppression, was ketamine sedation. Room change and cage changes were associated with minimal cortisol elevation and appetite suppression. Wild-born females showed more appetite suppression after room change than captive-born females. No differences were related to cage level. Pigtailed macaques strongly resembled longtailed macaques except they showed weaker responses to the new room and cage change, probably because the pigtails had spent more time in captivity. These findings support the conclusion that increasing cage size to the next regulation size category would not have measurable positive effects on the psychological well being of two species of laboratory macaques. PMID- 11051443 TI - Weaning, body weight, and postpartum amenorrhea duration in pigtailed macaques (Macaca nemestrina). AB - Early permanent infant separation or weaning decreases the time interval between pregnancies and interbirth intervals for many female primates. At least part of the interpregnancy interval consists of postpartum amenorrhea, a period of non menstruation lasting from the time of birth until the female begins to ovulate. This study investigated the effects of weaning age and dam's body weight on the duration of the interval between pregnancies, the duration of postpartum amenorrhea, and the number of cycles to conception in a year-round breeder. Female pigtailed macaques (Macaca nemestrina) have an observable perineal swelling that fluctuates throughout the menstrual cycle and provides a means of detecting ovulation. The perineal swelling records of socially housed pigtailed macaques were studied from July 1996 to September 1998. Postpartum amenorrhea data were obtained on 44 females who gave birth to normal, viable infants. As weaning age increased and dam's weight decreased, postpartum amenorrhea, and consequently the interval between pregnancies, increased in duration. The interpregnancy interval consisted almost entirely of the postpartum amenorrhea phase. Our finding that a higher dam's body weight decreased the length of postpartum amenorrhea duration lends support to the hypothesis that a minimum body weight is necessary for menstrual cycles to occur. Most females became pregnant on their first ovulation regardless of weaning age and whether or not they were carrying an infant. As the weaning age of the infant and the dam's weight increased, ovulation went from occurring after separation to occurring before separation. PMID- 11051444 TI - Techniques for collecting saliva from awake, unrestrained, adult monkeys for cortisol assay. AB - Cortisol levels serve as an index of pituitary-adrenal activity in nonhuman primates. In adult monkeys, cortisol is normally measured in blood (typically requiring restraint or sedation) or urine (reflecting a state rather than point estimate). In contrast, saliva collection is less invasive than drawing blood and allows for repeated sampling within a short period of time. Although protocols exist for collecting saliva from young monkeys, these procedures are inadequate for awake, unrestrained adult animals. Our laboratory has developed two methods for collecting saliva from adult rhesus monkeys: a "screen" method, which involves licking screen-covered gauze, and a "pole" method, which involves sucking and chewing on an attached rope. Twenty-three adult male rhesus monkeys were used to evaluate these two methods. After a period of adaptation, saliva samples were collected from 21 of 23 subjects. Saliva collection was faster with the pole than with the screen method (P < 0.01), but the pole method was not suitable for some animals because of their tendency to bite off the attached rope. An analysis of 19 saliva samples revealed a mean cortisol concentration of 0.84 microg/dl (range 0.27-1.77 microg/dl). There was no statistically significant difference in cortisol value between methods used (P > 0.22). The influence of the flavoring on the cortisol assay was tested, and was found to have no significant effect (P > 0.28). Our results indicate that either technique can be used to safely collect saliva from unrestrained adult monkeys. Choice of technique will depend on the proclivities of individual monkeys. PMID- 11051445 TI - Identification of monozygotic twin chimpanzees by microsatellite analysis. AB - Zygosity determination is important for epidemiological, biological, obstetric, and prognostic studies in both human and nonhuman primates. In this study, microsatellite loci were used to screen a pair of chimpanzee (Pan troglodytes) twins and their parents. The twins share identical alleles at all loci tested. The probability of dizygotic origin is estimated to be 2.9 x 10(-11). Even after excluding linkage of loci on the same chromosome, the probability is still low enough (3.7 x 10(-9)) to exclude dizygotic origin. MHC typing was also done on Patr-DRB and Patr-DQB loci and the twins share identical alleles at both loci, consistent with the microsatellite results. Together these results demonstrate a monozygotic origin for the chimp twins. Our results suggest that microsatellite analysis is a powerful method for zygosity determination, which can be screened reliably and efficiently. PMID- 11051446 TI - Observations on the birth and subsequent care of twin offspring by a lone pair of wild emperor tamarins (Saguinus imperator). AB - The birth of emperor tamarin (Saguinus imperator) twins was observed in the wild. The mother was a member of lone pair in a marked population of emperor tamarins in the Manu National Park, Peru. This report describes the birth and provides subsequent information on infant care and survival. Despite some difficulties, this lone pair of relatively young, primiparous emperor tamarins was able to successfully raise twin offspring to the age of at least 1 month. PMID- 11051447 TI - Dermatology practice management enhancement: implications for dermatology in the age of managed care. AB - Health care delivery in the United States has changed dramatically during the past 10 years. Dermatologists are faced with challenging changes in the way they learn new skills, practice, and provide dermatologic care. Dermatologists can survive and flourish in this environment if they learn proper management and enhancement skills. These skills include proper coding and documentation, regulatory compliance, and new levels of practice effectiveness and efficiency. Dermatologists can offer also the benefit of cosmetic procedures and ethical office-based dispensing to their patients. Greater future unification of this specialty will allow dermatology to flourish and show its need, efficiency, and cost-effectiveness. PMID- 11051448 TI - Dermatology practice management assures practice development and efficiency. AB - This article provides an overview of the disciplines involved in managing a dermatology practice today. Several key management processes, including strategic planning, financial analysis, advertising and public relations, information systems management, and compliance program development and monitoring are addressed. This article explores several possible tactics that can be used to help guide your practice in the right direction without overtaxing your resources. Also offered are possible solutions for creating an organization that is poised for success, and a management team capable of steering the practice through the sea of change ahead. PMID- 11051449 TI - Practice management for academic dermatology departments. AB - Practice management in the academic medical center (AMC) is different than in other environments. Practice is only a part of the practitioner's mission within an AMC. Practice revenue will be subject to a tax or overhead by both the school and the department. Contract and practice guidelines cannot be tailored to the needs of the dermatology practice, because contracts and guidelines are negotiated globally for all of the practices within the AMC. Personnel, on which the practice depends, may report to hospitals and clinics rather than to the practice's management. Even control of the practice's manager may be diluted by a dual or "dotted line" reporting relationship between the department manager and the school practice manager. Although more constraints exist within the AMC, there are some strategic and operational choices that affect a practice's success. Among these are: (1) selection of services offered; (2) creation of satellites; (3) stimulation of faculty effort; (4) enhancement of faculty billing knowledge; and (5) creation of a "tie" between staff and the practice. PMID- 11051450 TI - Choosing a computer billing system: avoid these mistakes. AB - Electronic billing systems make dermatology practices more efficient and productive. The decision to purchase and install such a system, however, involves a significant amount of time and is often accompanied by many mistakes. This article presents errors commonly made in the evaluation and purchase process and offers solutions. The success of dermatologists in the future will be partly determined by their effectiveness at managing information. PMID- 11051451 TI - Medicolegal issues for the dermatologist. AB - Medicolegal issues for the dermatologist cannot be ignored. Appropriate legal advice from an attorney with such expertise may represent an unwanted office expense. However, the value of this advice in an increasingly hostile health care environment cannot be underestimated. Legal consultation, an insurance policy against the vagaries of health care law, has become a necessity in our litigious society. PMID- 11051452 TI - Legal considerations in group practices and physician management. AB - Physicians are restructuring their practices in response to competition, managed care, and federal and state laws and regulations restricting their practice of medicine. Many physicians have formed group practices with the hope of selling this practice in the future. For physicians to profit from ancillary services, the group practice must meet federal and state requirements. If physicians choose to sell the group practice to a physician practice management company, careful review and negotiation of the purchase agreements are necessary to prevent complications if the arrangement is unsuccessful. PMID- 11051453 TI - Office dispensing: a responsible approach. AB - Office dispensing is a value added service in the dermatologist's office. As dermatologists we can recommend products with known scientific validity that will enhance patient care and provide reliable therapeutic results. Patients enjoy the convenience of being able to purchase products in the office and appreciate the dermatologist who spends time outlining a daily skin care regimen. Office dispensing benefits the physician by forcing him or her to keep current on new innovations in skin care and serves as an effective way to develop your cosmetic practice. Antiaging products, moisturizers and sunscreens, cleansers, and acne products are the basics for any in-office dispensing operation. The addition of hair and nail care products constitutes more advanced dispensing. Despite the mutual benefits for both the patient and physician, office dispensing continues to be controversial. The American Medical Association is concerned that selling health-related goods in the office may compromise the patient-physician relationship. The American Academy of Dermatology continues to support the right of dermatologists to dispense products in their office as long as it is in the best interest of the patient, as it is with all other dermatologic care. Dermatologists must preserve the right to dispense by conducting themselves in a highly professional and ethical manner. PMID- 11051454 TI - Will a physician assistant improve your dermatology practice? AB - A physician assistant (PA) is a licensed health care professional and dependent practitioner. The profession began in the 1960s and accredited programs now number 110 nationwide. PAs practice in every specialty, including dermatology, and their clinical duties vary tremendously. Research has shown enhanced productivity and increased patient satisfaction in practices using PAs. Most third-party payers cover services provided by PAs, making them ideally suited in this era of increasing managed care. PMID- 11051455 TI - Distribution of alphavbeta3, alphavbeta5 integrins and the integrin associated protein--IAP (CD47) in human glomerular diseases. AB - The alphav integrins present on the membrane of numerous cells, mediate attachment to matrix proteins, cell proliferation, migration and survival. We studied the expression of alphav integrinis and CD47 (a beta3 chain integrin associated protein) in various forms of glomerulonephritis (GN) characterized by mesangial proliferation and/or increased mesangial matrix. In normal glomeruli, epithelial cells expressed alphavbeta3, alphavbeta5 and CD47; endothelial cells expressed alpha5beta1 and CD47; mesangial cells expressed alphavbeta5, CD47, and to a less extent alphavbeta3. In acute post infectious GN (APIGN), membrano proliferative GN (MPGN) and diabetic nephropathy(DN), we observed that the beta3 chain, normally expressed by mesangial cells, was not detectable in the mesangium while its expression by epithelial cells was not modified. Parallel to the disappearance of alphavbeta3, the CD47 expression was decreased on the mesangial cells in MPGN, APIGN and DN. The expression of alphavbeta5 was clearly increased on podocytes and on proliferating mesangial cells in APIGN. By contrast, the mesangial expression of alphavbeta was normal or decreased in DN. The alpha5 chain of integrin, absent on normal mesangial cell, was expressed on proliferating mesangial cells in MPGN and APIGN. Thus, we observed modifications of alphavbeta3 and alphavbeta5 expression during human GN. The modulations of alphavbeta3 and alphavbeta5 expression differed according to the different glomerular cell types and were not parallel in glomerular cells: alphavbeta3 was decreased (and alphavbeta5 unchanged) on proliferating mesangial cells and alphavbeta5 was increased (and alphavbeta3 unchanged) in podocytes. This may reflect the existence of two distinct regulatory pathways. PMID- 11051457 TI - Co-localization of Rac1 and E-cadherin in human epidermal keratinocytes. AB - The Rac1 small GTP-binding protein is known to be involved in reorganization of the actin cytoskeleton and in regulation of intracellular signal transduction. The assembly and maintenance of cadherin-based cell cell junctions in epidermal keratinocytes is thought to be dependent on activity of Rac1. In this study we have generated green fluorescent protein (GFP)-tagged wild type, dominant negative and constitutively active Rac1 expression vectors and analyzed distribution of Rac1 following microinjection of human SCC12F epidermal keratinocytes. Wild type, dominant negative and constitutively active GFP Rac1 proteins distribute to sites of cell cell adhesion and co-localize with E cadherin and the catenins. Disruption of cadherin-based junctions by reduction in extracellular calcium concentrations, or by use of antibodies to E-cadherin, results in redistribution of Rac1 away from sites of cell cell interaction but the co-localization with E-cadherin is maintained. In addition, expression of constitutively active GFP Rac1 results in formation of membrane ruffles on the apical surface of cells and intracellular vesicles. Interestingly, co localization of Rac1 with E-cadherin is maintained in these structures. In contrast to previously published work we find that expression of dominant negative Rac1 neither disrupts cell cell adhesion nor prevents assembly of new cadherin-based adhesion structures. PMID- 11051456 TI - Soluble and cell-associated forms of the adhesion molecule LFA-3 (CD58) are differentially regulated by inflammatory cytokines. AB - The adhesion molecule lymphocyte function-associated antigen 3 (LFA-3) (CD58) is an important regulator of immune cell function which occurs as both surface associated and 'soluble' forms. This study has investigated the inter relationship and the effects of cytokines on the expression of LFA-3 isoforms. The surface antigen was found to be relatively unaffected by cytokines, but the release of soluble LFA-3 (sLFA-3) was highly responsive to interleukin 1beta (IL 1beta), interferon gamma (IFN-gamma) and tumour necrosis factor alpha (TNF alpha). This modulation was cell-specific, particularly with regard to IFN-gamma, which up-regulated sLFA-3 release by A431 cells but down-regulated the release of the soluble form from HEp2 and HepG2 cells. We further demonstrated that LFA-3 is also present in a cytoplasmic 'pool' in each of the cells and, moreover, that cleavage of LFA-3 from the cell surface by phospholipase C resulted in an increase in the levels of the intracellular LFA-3 and replacement of the membrane associated antigen. These observations suggest that the expression of the surface, soluble and intracellular forms of LFA-3 may be linked by regulatory mechanisms which are likely to exert an important influence on inflammatory interactions. PMID- 11051458 TI - Localization of urokinase type plasminogen activator to focal adhesions requires ligation of vitronectin integrin receptors. AB - Previous studies have shown that the adhesion protein, vitronectin, directs the localization of urokinase-type plasminogen activator (uPA) to areas of cell substrate adhesion, where uPA is thought to regulate cell migration as well as pericellular proteolysis. In the present study, HT-1080 cell lines expressing either wild-type vitronectin or vitronectin containing a single amino-acid substitution in the integrin binding domain were used to assess whether ligation of the alphavbeta5 integrin was required for uPA localization to focal adhesions. The synthesis of wild-type vitronectin by HT-1080 cells adherent to either collagen or fibronectin resulted in the redistribution of both the alphavbeta5 integrin as well as uPA to focal adhesion structures. In contrast, cells synthesizing mutant vitronectin, containing the amino-acid substitution in the integrin binding domain, were unable to direct the redistribution of either alphavbeta5 or uPA to focal adhesions. Recombinant forms of wild-type and mutant vitronectin were prepared in a baculovirus system and compared for their ability to direct the redistribution of vitronectin integrin receptors as well as uPA on human skin fibroblasts. In the absence of vitronectin, fibroblast cells adherent to fibronectin assemble focal adhesions which contain the beta1 integrin but do not contain uPA. Addition of recombinant wild-type, but not mutant, vitronectin to fibroblasts adherent to fibronectin resulted in the redistribution of alphavbeta3, alphavbeta5, and uPA into focal adhesions. However, when cells were plated directly onto antibodies directed against either the alphavbeta3 or alphavbeta5 integrins, uPA was not localized on the cell surface. These data indicate that ligation of vitronectin integrin receptors is necessary but not sufficient for the localization of uPA to areas of cell matrix adhesion, and suggest that vitronectin may promote cell migration by recruiting vitronectin integrin receptors and components of the plasminogen activator system to areas of cell matrix contact. PMID- 11051459 TI - Sialidase treatment exposes the beta1-integrin active ligand binding site on HL60 cells and increases binding to fibronectin. AB - The migration of neutrophils from the circulation to areas of inflammation is the result of the sequential activation of multiple cellular adhesion molecules. beta1-Integrins are cell surface glycoproteins and the class of adhesion molecules responsible for binding to the extracellular matrix. The goal of this study was to determine the contribution of glycosylation, specifically the presence of sialic acid, to beta1-integrin adhesion in a neutrophil model. beta1 Integrins on differentiated HL60 cells were remodeled by treatment with the exoglycosidases, sialidase and beta-galactosidase. beta1-Integrin activity was determined by measuring adherence to the extracellular matrix protein fibronectin. The expression of beta1-integrins, beta2-integrins and activated beta1-integrins was determined by flow cytometry. Remodeling of beta1-integrins by treatment with sialidase increased adhesion by greater than 1,000%. Flow cytometric analysis of remodeled beta1-integrins demonstrated an increased expression of the activated beta1-integrin, but only minor increases in the expression of total beta1- and beta2-integrins. We postulate that glycosidase treatment increases adhesion and expression of activated beta1-integrins by exposure of the normally hidden ligand-binding site. The glycosylation of beta1 integrins on neutrophils may act to hide the ligand-binding site in unstimulated cells thereby contributing to the affinity modulation observed in neutrophil beta1-integrin function. PMID- 11051460 TI - Quantitative determination of gap junction intercellular communication using flow cytometric measurement of fluorescent dye transfer. AB - Gap junction intercellular communication (GJIC) is involved in several aspects of normal cell behaviour, and disturbances in this type of communication have been associated with many pathological conditions. Reliable and accurate methods for the determination of GJIC are therefore important in studies of cell biology. (Tomasetto, C., Neveu, M.J., Daley, J., Horan, P.K. and Sager, R. (1993) Journal of Cell Biology, 122, 157-167) reported some years ago the use of flow cytometer to determine transfer between cells of a mobile dye, calcein, as a measure of cell communication through gap junctions. In spite of this being a method with potential for quantitative and reliable determination of GJIC, it has been modestly used, possibly due to technical difficulties. In the present work we have illustrated several ways to use flow cytometric data to express cell communication through gap junctions. The recipient cells were pre-stained with the permanent lipophilic dye PKH26, and the donor cell population were loaded with the gap junction permeable dye, calcein. We show that the method may be used to measure the effect of chemicals on GJIC, and that the information is reliable, objective and reproducible due to the large number of cells studied. The data may give additional information to that obtained with other methods, since the effect observed will be on the establishment of cell communication as compared to what is observed for microinjection or scrape loading, where the effect is on already established communication. This is probably the reason for the more potent effects of DMSO on GJIC measured by the present method than on already existing GJIC measured by microinjection or quantitative scrape loading. We also show that the problem related to the mobile dye calcein not being fixable with aldehydes will not affect the results as long as the cells are kept on ice in the dark and analysed by flow cytometer within the first hours after formalin cell fixation. PMID- 11051461 TI - Modulation of heart fibroblast migration and collagen gel contraction by IGF-I. AB - Dynamic interactions between cells and the extracellular matrix are essential in the regulation of a number of cellular processes including migration, adhesion, proliferation and differentiation. A variety of factors have been identified which modulate these interactions including transforming growth factor-beta, platelet-derived growth factor and others. Insulin-like growth factors have been shown to regulate collagen production by heart fibroblasts; however, the effects of this growth factor on the interactions of heart fibroblasts with the extracellular matrix have not been examined. The present studies were carried out to determine the effects of IGF-I on the ability of fibroblasts to interact with the extracellular matrix and to begin to determine the mechanisms of this response. These experiments illustrate that IGF-I treatment results in increased migration, collagen reorganization and gel contraction by heart fibroblasts. IGF I has been shown to activate both the mitogen-activated protein kinase and phophatidylinositol-3 kinase pathways in isolated cells. Experiments with pharmacological antagonists of these pathways indicate that the mitogen-activated protein kinase pathway is essential for IGF-I stimulated collagen gel contraction by fibroblasts. These studies illustrate that IGF-I modulates the ability of fibroblasts to interact with the collagen matrix and that activation of multiple signaling pathways by IGF-I may produce distinct downstream responses in these cells. PMID- 11051462 TI - New fellows of the Acoustical Society of America. PMID- 11051463 TI - ASA North Texas Chapter presents awards at 43rd Dallas Morning News-Toyota Regional Science and Engineering Fair. PMID- 11051465 TI - Localization of a pontine vocalization-controlling area. AB - To find out whether there exist additional regions in the pontine brainstem, apart from the phonatory motoneuron pools involved in vocal motor control, the effects of a localized blockade of excitatory neurotransmission in the pons were studied on squirrel monkey vocalization. Vocalization was elicited by electrical stimulation of the periaqueductal gray of the midbrain. Blockade was carried out by stereotaxic injections of kynurenic acid, a nonspecific glutamate antagonist. It was found that injections made into the ventrolateral pons around the ventral nucleus of the lateral lemniscus and superior olive could block periaqueductally elicited vocalization. Injections were only effective ipsilaterally, not contralaterally to the stimulation site. The blockade was limited to one particular class of calls, all of which had in common a characteristic stereotyped frequency modulation over several kHz. It is concluded that critical processing steps of vocal motor control take place in the periolivary region. PMID- 11051466 TI - Resonance properties of the vocal folds: in vivo laryngoscopic investigation of the externally excited laryngeal vibrations. AB - The study presents the first attempt to investigate resonance properties of the living vocal folds by means of laryngoscopy. Laryngeal vibrations were excited via a shaker placed on the neck of a male subject and observed by means of videostroboscopy and videokymography (VKG). When the vocal folds were tuned to the phonation frequency of 110 Hz and sinusoidal vibration with sweeping frequency (in the range 50-400 Hz) was delivered to the larynx, three clearly pronounced resonance peaks at frequencies around 110, 170, and 240 Hz were identified in the vocal fold tissues. Different modes of vibration of the vocal folds, observed as distinct lateral-medial oscillations with one, two, and three half-wavelengths along the glottal length, respectively, were associated with these resonance frequencies. At the external excitation frequencies below 100 Hz, vibrations of the ventricular folds, aryepiglottic folds and arytenoid cartilages were dominant in the larynx. PMID- 11051467 TI - Time normalization of voice signals using functional data analysis. AB - The harmonics-to-noise ratio (HNR) has been used to quantify the waveform irregularity of voice signals [Yumoto et al., J. Acoust. Soc. Am. 71, 1544-1550 (1982)]. This measure assumes that the signal consists of two components: a harmonic component, which is the common pattern that repeats from cycle-to-cycle, and an additive noise component, which produces the cycle-to-cycle irregularity. It has been shown [J. Qi, J. Acoust. Soc. Am. 92, 2569-2576 (1992)] that a valid computation of the HNR requires a nonlinear time normalization of the cycle wavelets to remove phase differences between them. This paper shows the application of functional data analysis to perform an optimal nonlinear normalization and compute the HNR of voice signals. Results obtained for the same signals using zero-padding, linear normalization, and dynamic programming algorithms are presented for comparison. Functional data analysis offers certain advantages over other approaches: it preserves meaningful features of signal shape, produces differentiable results, and allows flexibility in selecting the optimization criteria for the wavelet alignment. An extension of the technique for the time normalization of simultaneous voice signals (such as acoustic, EGG, and airflow signals) is also shown. The general purpose of this article is to illustrate the potential of functional data analysis as a powerful analytical tool for studying aspects of the voice production process. PMID- 11051468 TI - Frication noise modulated by voicing, as revealed by pitch-scaled decomposition. AB - A decomposition algorithm that uses a pitch-scaled harmonic filter was evaluated using synthetic signals and applied to mixed-source speech, spoken by three subjects, to separate the voiced and unvoiced parts. Pulsing of the noise component was observed in voiced frication, which was analyzed by complex demodulation of the signal envelope. The timing of the pulsation, represented by the phase of the anharmonic modulation coefficient, showed a step change during a vowel-fricative transition corresponding to the change in location of the noise source within the vocal tract. Analysis of fricatives [see text] demonstrated a relationship between steady-state phase and place, and f0 glides confirmed that the main cause was a place-dependent delay. PMID- 11051469 TI - Nonlinear phenomena in the natural howling of a dog-wolf mix. AB - It was reported to the first author that a female dog-wolf mix showed anomalously rough-sounding vocalization. Spectral analysis of recordings of the vocalization revealed frequency occurrences of subharmonics, biphonation (two independent pitches) and chaos. Since these nonlinear phenomena are currently widely discussed as integral to mammalian vocalization [Wilden et al., Bioacoustics 9, 171-196 (1988)] or as indicators of vocal pathologies [Herzel et al., J. Speech Hearing Res. 37, 1008-1019 (1994); Riede et al., Z. Sgtkde 62 Suppl: 198-203 (1997)], we sought to understand the production mechanism of the observed vocal instabilities. First the frequency of nonlinear phenomena in the calls was determined for the female and four additional individuals. It turned out that these phenomena appear, but much less frequently in the repertoire of the four other animals. The larynges of the female and two other individuals were dissected post mortem. There was no apparent asymmetry of the vocal folds but a slight asymmetry of the arytenoid cartilages. The most pronounced difference, however, was an upward extension of both vocal folds of the female. This feature is reminiscent of "vocal lips" (syn. "vocal membranes") in some primates and bats. Spectral analysis of the female's voice showed clear similarities with an intensively studied voice of a human who produces biphonation intentionally. Finally, the possible communicative relevance of nonlinear phenomena is discussed. PMID- 11051470 TI - Vocalization-correlated respiratory movements in the squirrel monkey. AB - Respiratory abdominal movements associated with vocalization were recorded in awake squirrel monkeys. Several call types, such as peeping, trilling, cackling, and err-chucks, were accompanied by large vocalization-correlated respiratory movements (VCRM) that started before vocalization. During purring, in contrast, only small VCRM were recorded that started later after vocal onset. VCRM during trill calls, a vocalization type with repetitive frequency modulation, showed a modulation in the rhythm of the frequency changes. A correlation with amplitude modulation was also present, but more variable. As high frequencies need a higher lung pressure for production than low frequencies, the modulation of VCRM seems to serve to optimize the lung pressure in relation to the vocalization frequency. The modulation, furthermore, may act as a mechanism to produce different trill variants. During err-chucks and staccato peeps, which show a large amplitude modulation, a nonmodulated VCRM occurred. This indicates the existence of a laryngeal amplitude-controlling mechanism that is independent from respiration. PMID- 11051471 TI - Interior near-field acoustical holography in flight. AB - In this paper boundary element methods (BEM) are mated with near-field acoustical holography (NAH) in order to determine the normal velocity over a large area of a fuselage of a turboprop airplane from a measurement of the pressure (hologram) on a concentric surface in the interior of the aircraft. This work represents the first time NAH has been applied in situ, in-flight. The normal fuselage velocity was successfully reconstructed at the blade passage frequency (BPF) of the propeller and its first two harmonics. This reconstructed velocity reveals structure-borne and airborne sound-transmission paths from the engine to the interior space. PMID- 11051472 TI - A conjugated infinite element method for half-space acoustic problems. AB - Many acoustic problems (especially in environmental acoustics) involve half-space domains bounded by a plane subjected to normal admittance boundary conditions. In the "low" frequency domain, the numerical treatment of such problems usually relies on boundary element methods based on a particular Green's function suited for the half-(admittance) plane. In the present paper, an alternative hybrid finite/infinite element scheme is proposed. The method relies on a direct treatment of nonhomogeneous boundary conditions along infinite element edges (or faces). The procedure is validated through comparisons with an available reference solution. PMID- 11051473 TI - Backscattering cross section of a rigid biconic reflector. AB - Plane wave incidence on a rigid biconic target is considered. A biconic reflector consists of two cones truncated by planes perpendicular to their axes and joined at their smaller flat faces. The cone angles are allowed to be variable, provided their sum is equal to 90 degrees. The backscattering cross section is expressed in terms of a surface integral of the geometrical acoustics field, which results from incident singly and doubly reflected rays. A saddle-point calculation gives a first-order high-frequency approximation in which the backscattering cross section is proportional to the incident wave number and a function of the angle of incidence, cone angles, and inner and outer radii of the truncated cones. This expression is algebraically complex but easy to implement numerically. Results are presented that exercise the parameters of the problem. An interesting result of the solution is that for fixed outer radii there is a nonzero optimum inner radius for backscattering strength. For broadside incidence on 45 degrees cones with equal outer radii, this optimum value is approximately equal to 11% of the outer radius. PMID- 11051474 TI - Complex modal statistics in a reverberant dissipative body. AB - The statistics of the ultrasonic resonance peaks of a finite elastic body are investigated. The distribution of peak phases, and the normalized variance of peak amplitudes, are shown to be consistent with a hypothesis that the modes themselves are complex Gaussian random numbers. A value q = 0.33 for the ratio of the standard deviations of the imaginary and real parts of the modes is found to fit the data, and to bring recent theory of power variances into better accord with measurements. PMID- 11051475 TI - Experimental study of sound propagation in a chain of spherical beads. AB - In this paper are described experimental observations which are concerned by the propagation of pulsed ultrasonic waves transmitted through a limited one dimensional periodic granular medium submitted to a static force. This study- which is limited to a time domain analysis--exhibits experimental results which depend on the polarization of the acoustic excitation. In the case of compressional excitation, spherical Rayleigh type surface waves propagate around the beads. In the case of shear excitation, the experimental recordings point out the existence of a very low signal, the frequency of which is equal to the cut off frequency of the chain. Moreover it is established that the frequency value varies with the radius of the bead, the normal force applied to the beads, and the mechanical properties of the material. PMID- 11051476 TI - Analyses of axisymmetric waves in layered piezoelectric rods and their composites. AB - An exact treatment of the propagation of axisymmetric waves in coaxial anisotropic assembly of piezoelectric rod systems is presented. The rod system consists of an arbitrary number of coaxial layers, each possessing transversely isotropic symmetry properties. The treatment, which is based on the transfer matrix technique, is capable of deriving the dispersion relations for a variety of situations. These include the case of a single rod system that is either embedded in an infinitely extended solid or fluid host or kept free. The procedure is also adapted to derive approximate solutions for the cases of a periodic fiber distribution in a matrix material, which model unidirectional fiber-reinforced composites. The results are numerically illustrated for a widely used piezoelectric-polymer composite. It is seen that piezoelectric coupling can significantly change the morphology of the dispersive behavior of the composite. PMID- 11051477 TI - Second-harmonic generation in a sound beam reflected and transmitted at a curved interface. AB - This article presents a model for second-harmonic generation in a sound beam that is reflected from or transmitted through a curved interface. Propagation in homogeneous fluids is assumed. Simple analytic solutions are derived for the case of focused Gaussian beams. The solutions are used to illustrate the effects of focusing or defocusing due to curvature of the interface, in combination with impedance change at the interface, on energy transfer from the fundamental to the second harmonic. PMID- 11051478 TI - Compressibility effects on steady streaming from a noncompact rigid sphere. AB - The problem of steady streaming around a rigid isolated sphere in a plane standing acoustic field is considered. Existing results in the literature have been generalized to allow for noncompactness of the sphere, and the influence of fluid compressibility on the streaming behavior has been included. It is found that in the high-frequency limit of interest for which the streaming is strongest, the effective steady slip velocity at the edge of the inner boundary layer region that is responsible for driving the steady streaming in the bulk of the fluid in the outer region, has a complex variation over the surface of the sphere that depends on (i) the sphere position (with respect to the node/antinode of the acoustic field), (ii) the extent of sphere compactness, and (iii) on a well-defined function (representing compressibility effects) of the fluid Prandtl number and its ratio of specific heats. Not surprisingly, the contribution from this function is negligible when the host fluid is a liquid. The steady streaming behavior around the sphere is demonstrated with the help of flow streamlines for various cases in the diffusive limit of weak outer flow for low streaming Reynolds numbers. PMID- 11051479 TI - Condensation in a steady-flow thermoacoustic refrigerator. AB - Condensation may occur in an open-flow thermoacoustic cooler with stack temperatures below the saturation temperature of the flowing gas. In the experimental device described here the flowing gas, which is also the acoustic medium, is humid air, so the device acts as a flow-through dehumidifier. The humid air stream flows through an acoustic resonator. Sound energy generated by electrodynamic drivers produces a high-amplitude standing wave inside of the resonator, which causes cooling on a thermoacoustic stack. Condensation of water occurs as the humid air passes through the stack and is cooled below its dew point, with the condensate appearing on the walls of the stack. The dry, cool air passes out of the resonator, while the condensate is wicked away from the end of the stack. Thermoacoustic heat pumping is strongly affected by the form of the condensate inside of the stack, whether condensed mostly on the stack plates, or largely in the form of droplets in the gas stream. Two simple models of the effect of the condensate are matched to a measured stack temperature profile; the results suggest that the thermoacoustic effect of droplets inside the stack is small. PMID- 11051480 TI - Fluctuations of spherical waves in a turbulent atmosphere: effect of the axisymmetric approximation in computational methods. AB - The validity of the axisymmetric parabolic-equation (PE) method for line-of-sight sound propagation in a turbulent atmosphere is investigated. The axisymmetric PE method is a finite-difference method for solving a 2D parabolic wave equation, which follows from the 3D wave equation by the assumption of axial symmetry around the vertical axis through the source. It is found that this axisymmetric approximation has a considerable spurious effect on the fluctuations of the sound field. This is concluded from analytical expressions for the log-amplitude and phase variances, derived both for isotropic turbulence and for axisymmetric turbulence. The expressions for axisymmetric turbulence are compared with the results of numerical computations with the PE method. PMID- 11051481 TI - Variability in the passive ranging of acoustic sources in air using a wavefront curvature technique. AB - Simultaneous time delay measurements from two adjacent pairs of sensors are used to estimate the "instantaneous" range and bearing of both stationary and moving broadband sources of continuous sound in air. The random fluctuations in the estimated source position are due to random errors in the time delay measurements. The variances of both the short time-scale and long time-scale random errors in the time delay estimates are quantified for different sensor separation distances and for different source positions. After normalization, the observed variances of the short time-scale random errors in the bearing and range estimates are found to agree with the theoretical results predicted for various sensor-source configurations where the independent variable is the length of the effective intersensor baseline. Increasing the intersensor separation distance by an order of magnitude reduces the bearing error variance by two orders of magnitude and the range error variance by four orders of magnitude. PMID- 11051482 TI - The effect of temperature on sound wave absorption in a sediment layer. AB - The effect of temperature on sound velocity, absorption, and reflection coefficient in the seabed sediment layer is investigated. Experimental measurements of sound speed, absorption, and the reflection coefficient in a sandy sediment layer have been carried out at several temperatures. An absorption reduction of 75 dB/m and a velocity increase of 65 m/s have been measured at a frequency of 1 MHz when the temperature increases from 5 to 25 degrees C. Because of the absorption temperature dependence the amplitude of the reflected wave from the back surface of the sub-bottom layer after going back and forth across the layer increases with the temperature. PMID- 11051483 TI - An optimization approach to multi-dimensional time domain acoustic inverse problems. AB - An optimization approach to a multi-dimensional acoustic inverse problem in the time domain is considered. The density and/or the sound speed are reconstructed by minimizing an objective functional. By introducing dual functions and using the Gauss divergence theorem, the gradient of the objective functional is found as an explicit expression. The parameters are then reconstructed by an iterative algorithm (the conjugate gradient method). The reconstruction algorithm is tested with noisy data, and these tests indicate that the algorithm is stable and robust. The computation time for the reconstruction is greatly improved when the analytic gradient is used. PMID- 11051484 TI - Spectral integral representations of volume scattering in sediments in layered waveguides. AB - In situ measurements of scattering strength are often obtained by analyzing the early-time, high-angle reverberation from bottom and subbottom features. In order to provide insight into the mechanisms which cause bottom reverberation, and to their distinguishing characteristics, it is necessary to have a capability for modeling both the rough surface and the volume scattering mechanisms. For high angle, early-time backscatter, the most appropriate approach is to use a spectral integral representation, which naturally includes the continuous spectrum important for this angular regime. A rough surface scattering theory developed earlier in this framework has provided important insights into wave scattering and penetration physics at the seafloor. Here a consistent representation for the subbottom scattering is developed and examples are provided which illustrate the observable differences between the two scattering mechanisms. PMID- 11051485 TI - Acoustic measurements of boundary layer flow and sediment flux. AB - Results are reported on an assessment of the application of coherent Doppler and cross-correlation techniques to measure nearbed boundary layer flow. The approaches use acoustic backscattering from sediments entrained into the water column from the bed, to obtain high-resolution profiles of the nearbed hydrodynamics. Measurements are presented from a wave tunnel experiment in which sediment was entrained by unidirectional, oscillatory, and combined flows. The data collected have been used to evaluate the capability of the two flow techniques to measure boundary layer mean, turbulent, and intrawave velocity profiles. Further, the backscattered signal has been used to measure suspended sediment concentration profiles, which have been combined with the velocity profiles to obtain high-resolution measurements of boundary layer sediment flux. PMID- 11051486 TI - Source ranging with minimal environmental information using a virtual receiver and waveguide invariant theory. AB - A method is presented for estimating the range of an unknown broadband acoustic source in a waveguide, using a vertical array and a signal sample from another broadband source at a known location relative to the array. The method requires no modeling of the acoustic field, and little to no environmental information for flat bathymetries. Waveguide invariant theory [e.g., D'Spain and Kuperman, J. Acoust. Soc. Am 106, 2454-2468 (1999)] is applied to the "virtual receiver" [Siderius et al., J. Acoust. Soc. Am. 102, 3439-3449 (1997)] to create a "virtual aperture" (VA). In effect, the method effectively converts a source at known range r(g) into a continuum of receivers lying between ranges (1 +/- alpha/beta)*r(g), where beta is a scalar parameter called the acoustic invariant, and alpha approximately 0.1. This effective displacement is achieved by correlating the known source field, measured at frequency component omega, with the unknown source field, measured at frequency component omega + omegas. When the VA output is plotted as a function of omega and omegas, the slope of the resulting correlation contours yields the unknown source range. The concept is illustrated via both simulation and analysis of data collected from a pseudo random noise source with 75-150-Hz bandwidth during SWellEx-3, a shallow water experiment conducted off the San Diego coast. The virtual aperture can be reformulated for range-dependent environments, if adiabatic propagation assumptions are valid, and if the bathymetry surrounding the array is known. PMID- 11051487 TI - Backscatter of high-frequency (200kHz) acoustic wavefields from ocean turbulence. AB - Near space and time coincident 200-kHz acoustic backscatter and CTD measurements were taken during an interdisciplinary study of the internal wave packets that propagate through Massachusetts Bay. The data strongly support the contention that acoustic wavefields can be backscattered from turbulent mixing events (microstructure) associated with the internal wave packets. PMID- 11051488 TI - An analysis of Freedman's "image pulse" model in air. AB - The "image pulse" model developed by Freedman calculates the echoes generated from convex objects in an underwater environment after insonification with a narrow-band transient signal. The model uses the source radiation and the solid angle subtended at the transducer by the scattering body to determine the echo structure. Work has been completed in adapting this model for use in an air environment using noncoincident transmitters and receivers. Experiments were conducted to measure the amplitudes of the echoes off a range of convex objects, at distances up to 1.4 m, after insonification with a Polaroid transducer. These measured amplitudes were compared to those predicted by the model, with the results for cones highlighting the limitations of the model. Spheres, however, performed significantly better, with an average error of under 5%, indicating that the model should be reasonably accurate at calculating the echoes off convex objects with a smoothly varying surface. PMID- 11051489 TI - Extension of the mode method for viscoelastic media and focused ultrasonic beams. AB - In the present study viscoelasticity is introduced in the mode model and the orthogonality condition is adapted for viscous media. The expansion of convergent acoustic Gaussian beams in terms of radiation modes for viscoelastic media is studied as well. The effects on the reflected and transmitted profiles of acoustic beams incident from an ideal liquid onto a viscoelastic plate are shown and physically explained. It is shown that focusing the incident beam can suppress divergence effects and gives the possibility to measure shear wave attenuation coefficients. PMID- 11051490 TI - Bounded beam interaction with thin inclusions. Characterization by phase differences at Rayleigh angle incidence. AB - A theoretical study of the reflection of a bounded Gaussian ultrasonic beam, incident onto a rectangular inclusion located near a fluid/solid half-space interface, is presented. The thickness of the inclusion is assumed to be much smaller than the ultrasonic wavelength in the solid. It is shown that, at critical Rayleigh angle incidence, the phase in the point of maximum amplitude of the shifted reflected lobe is very sensitive to dimension variations of the inclusion, and thus useful for inclusion characterization. The modelization of the problem is based on mode theory. PMID- 11051491 TI - Fan beam and double crosshole Lamb wave tomography for mapping flaws in aging aircraft structures. AB - As the worldwide aviation fleet continues to age, methods for accurately predicting the presence of structural flaws-such as hidden corrosion and disbonds that compromise airworthiness become increasingly necessary. Ultrasonic guided waves, Lamb waves, allow large sections of aircraft structures to be rapidly inspected. However, extracting quantitative information from Lamb wave data has always involved highly trained personnel with a detailed knowledge of mechanical waveguide physics. The work summarized here focuses on a variety of different tomographic reconstruction techniques to graphically represent the Lamb wave data in quantitative maps that can be easily interpreted by technicians. Because the velocity of Lamb waves depends on thickness, for example, the traveltimes of the fundamental Lamb modes can be converted into a thickness map of the inspection region. This article describes two potentially practical implementations of Lamb wave tomographic imaging techniques that can be optimized for in-the-field testing of large-area aircraft structures. Laboratory measurements discussed here demonstrate that Lamb wave tomography using either a ring of transducers with fan beam reconstructions, or a square array of transducers with algebraic reconstruction tomography, is appropriate for detecting flaws in multilayer aircraft materials. The speed and fidelity of the reconstruction algorithms as well as practical considerations for person-portable array-based systems are discussed in this article. PMID- 11051492 TI - Dynamics of gas bubbles in viscoelastic fluids. II. Nonlinear viscoelasticity. AB - The nonlinear oscillations of a spherical, acoustically forced gas bubble in nonlinear viscoelastic media are examined. The constitutive equation [Upper Convective Maxwell (UCM)] used for the fluid is suitable for study of large amplitude excursions of the bubble, in contrast to the previous work of the authors which focused on the smaller amplitude oscillations within a linear viscoelastic fluid [J. S. Allen and R. A. Roy, J. Acoust. Soc. Am. 107, 3167-3178 (2000)]. Assumptions concerning the trace of the stress tensor are addressed in light of the incorporation of viscoelastic constitutive equations into bubble dynamics equations. The numerical method used to solve the governing system of equations (one integrodifferential equation and two partial differential equations) is outlined. An energy balance relation is used to monitor the accuracy of the calculations and the formulation is compared with the previously developed linear viscoelastic model. Results are found to agree in the limit of small deformations; however, significant divergence for larger radial oscillations is noted. Furthermore, the inherent limitations of the linear viscoelastic approach are explored in light of the more complete nonlinear formulation. The relevance and importance of this approach to biomedical ultrasound applications are highlighted. Preliminary results indicate that tissue viscoelasticity may be an important consideration for the risk assessment of potential cavitation bioeffects. PMID- 11051493 TI - Analysis and comparison of four anhysteretic polarization models for lead magnesium niobate. AB - Four anhysteretic polarization models that have been used in the literature to evaluate data acquired from lead magnesium niobate (PMN) are analyzed and compared. Derivations of two of the models from assumed spatial distributions of dipole energy states, using first physical principles, are presented. A third model is derived from a suitable integral averaging calculation. These derivations are used as the basis for developing an integral equation for determining an energy-state distribution that produces a fourth model, which was not originally formulated in terms of an assumed distribution. A new polarization function is also presented. Excellent approximations to each of the four polarization functions of interest can be deduced from this new polarization function by adjusting the numerical value of just a single parameter. An application of two of the models to data is presented. It is shown that it can be necessary to consider a sample to be an admixture of two distinct species of poles, in the sense that two polarization functions must be added together in order to accommodate the data. PMID- 11051494 TI - An efficient model of an equipment loaded panel for active control design studies. AB - An effective investigation of alternative control strategies for the reduction of vibration levels in satellite structures requires realistic, yet efficient, structural models to simulate the dynamics of the system. These models should include the effects of the sources, receivers, supporting structure, sensors, and actuators. In this paper, a modeling technique which meets these requirements is developed and some active control strategies are briefly investigated. The particular subject of investigation is an equipment-loaded panel and the equations of motion are derived using the Lagrange-Rayleigh-Ritz (LRR) approach. The various pieces of equipment on the panel are mounted on active or passive suspensions, and resonators are used to represent the internal dynamics of the mounted equipment. Control of the panel, which transmits vibrations from sources to receivers, is by means of piezoelectric patches and the excitation consists of dynamic loads acting on the equipment enclosures and/or directly on the panel. The control objective is to minimize the displacement at an arbitrary output location. The LRR model developed is verified against one produced by using the finite-element method. Finally, some initial controller design studies are undertaken to investigate and compare the effectiveness of different control strategies (e.g., minimization at the source, along the vibration path, or at the receiver). PMID- 11051495 TI - Estimation of structural wave numbers from spatially sparse response measurements. AB - A method is presented for estimating the complex wave numbers and amplitudes of waves that propagate in damped structures, such as beams, plates, and shells. The analytical basis of the method is a wave field that approximates response measurements in an aperture where no excitations are applied. At each frequency, the method iteratively adjusts wave numbers to best approximate response measurements, using wave numbers at neighboring frequencies as initial estimates in the search. In comparison to existing methods, the method generally requires far fewer measurement locations and does not require evenly spaced locations. The number of locations required by the method scales with the number of waves that propagate in the structure, whereas the number of locations required by existing methods scales with the minimum wavelength. In addition, the method allows convenient inclusion of the analytic relationships between wave numbers that exist for flexural vibrations of beams and plates. Advantages of the method are illustrated by an example in which a beam is excited by a transverse force at one end. Using analytic data and experimental measurements, the method produces a wave field that matches response measurements to within 1 percent. One interesting feature of the new method is that, when applied to analytic data, it supplies more robust wave number estimates using responses at unevenly spaced locations. PMID- 11051496 TI - Acoustic noise during functional magnetic resonance imaging. AB - Functional magnetic resonance imaging (fMRI) enables sites of brain activation to be localized in human subjects. For studies of the auditory system, acoustic noise generated during fMRI can interfere with assessments of this activation by introducing uncontrolled extraneous sounds. As a first step toward reducing the noise during fMRI, this paper describes the temporal and spectral characteristics of the noise present under typical fMRI study conditions for two imagers with different static magnetic field strengths. Peak noise levels were 123 and 138 dB re 20 microPa in a 1.5-tesla (T) and a 3-T imager, respectively. The noise spectrum (calculated over a 10-ms window coinciding with the highest-amplitude noise) showed a prominent maximum at 1 kHz for the 1.5-T imager (115 dB SPL) and at 1.4 kHz for the 3-T imager (131 dB SPL). The frequency content and timing of the most intense noise components indicated that the noise was primarily attributable to the readout gradients in the imaging pulse sequence. The noise persisted above background levels for 300-500 ms after gradient activity ceased, indicating that resonating structures in the imager or noise reverberating in the imager room were also factors. The gradient noise waveform was highly repeatable. In addition, the coolant pump for the imager's permanent magnet and the room air handling system were sources of ongoing noise lower in both level and frequency than gradient coil noise. Knowledge of the sources and characteristics of the noise enabled the examination of general approaches to noise control that could be applied to reduce the unwanted noise during fMRI sessions. PMID- 11051497 TI - Sound absorption of cellular metals with semiopen cells. AB - A combined experimental and theoretical study is presented for the feasibility of using aluminum foams with semiopen cells for sound-absorption applications. The foams are processed via negative-pressure infiltration, using a preform consisting of water-soluble spherical particles. An analytical model is developed to quantify the dependence of pore connectivity on processing parameters, including infiltration pressure, particle size, wetting angle, and surface tension of molten alloy. Normal sound-absorption coefficient and static flow resistance are measured for samples having different porosity, pore size, and pore opening. A theory is developed for idealized semiopen metallic foams, with a regular hexagonal hollow prism having one circular aperture on each of its eight surfaces as the unit cell. The theory is built upon the acoustic impedance of the circular apertures (orifices) and cylindrical cavities due to viscous effects, and the principle of electroacoustic analogy. The predicted sound-absorption coefficients are compared with those measured. To help select processing parameters for producing semiopen metallic foams with desirable sound-absorbing properties, emphasis is placed on revealing the correlation between sound absorption and morphological parameters such as pore size, pore opening, and porosity. PMID- 11051498 TI - Surface diffusion coefficients for room acoustics: free-field measures. AB - A surface diffusion coefficient is needed in room acoustics to enable the quality of diffusing surfaces to be evaluated. It may also facilitate more accurate geometric room acoustic models. This paper concentrates on diffusion coefficients derived from free-field polar responses. An extensive set of two- and three dimensional measurements and predictions was used to test the worth of different diffusion coefficient definitions. The merits and problems associated with these types of coefficients are discussed, and past parameters reviewed. Two new coefficients are described. The new measure based on the autocorrelation function is forwarded as the best free-field coefficient. The strengths and weaknesses of the coefficient are defined. PMID- 11051499 TI - Predicting reverberation times in a simulated classroom. AB - By varying the sound-absorption treatments in a simulated classroom, experimental results were compared with analytical and computer predictions of reverberation time. Analytical predictions were made with different absorption exponents, which are the result of different weighting procedures involving room surface areas and the sound-absorption coefficients. Sound scattering was found to influence measured reverberation times. With the amount of sound scattering provided, more accurate analytical predictions were obtained with absorption exponents that give reverberation times longer than those obtained with the Sabine absorption exponent, which consistently underpredicted reverberation times. However, none of the absorption exponents could be singled out as more adequate because of similar average accuracy. Computer predictions of reverberation time were accomplished with two commercially available ray-based programs, RAYNOISE 3.0 and ODEON 2.6, with specular and calibrated diffuse reflection procedures. Neither type of procedure, in either program, was more accurate than the best analytical predictions. With RAYNOISE, neither the specular nor the calibrated diffuse reflection procedure could be singled out as more adequate. For ODEON, the calibrated diffuse reflection procedure gave consistently more accurate predictions than its specular reflection procedure, with the best accuracy of the computer predictions. PMID- 11051500 TI - Comparison of an integral equation on energy and the ray-tracing technique in room acoustics. AB - This paper deals with a comparison of two room acoustic models. The first one is an integral formulation stemming from power balance and the second is the ray tracing technique with a perfectly diffuse reflection law. The common assumptions to both models are the uncorrelated wave hypothesis and the perfectly diffuse reflection law. The latter allows the use of these methods for nondiffuse fields beyond the validity domain of Sabine's formula. Comparisons of numerical simulations performed with the softwares RAYON and CeReS point out that these results are close to each other and finally, a formal proof is proposed showing that both methods are actually equivalent. PMID- 11051501 TI - Application of a finite-element model to low-frequency sound insulation in dwellings. AB - The sound transmission between adjacent rooms has been modeled using a finite element method. Predicted sound-level difference gave good agreement with experimental data using a full-scale and a quarter-scale model. Results show that the sound insulation characteristics of a party wall at low frequencies strongly depend on the modal characteristics of the sound field of both rooms and of the partition. The effect of three edge conditions of the separating wall on the sound-level difference at low frequencies was examined: simply supported, clamped, and a combination of clamped and simply supported. It is demonstrated that a clamped partition provides greater sound-level difference at low frequencies than a simply supported. It also is confirmed that the sound-pressure level difference is lower in equal room than in unequal room configurations. PMID- 11051502 TI - Detection of Gaussian signals in Poisson-modulated interference. AB - Passive broadband detection of target signals by an array of hydrophones in the presence of multiple discrete interferers is analyzed under Gaussian statistics and low signal-to-noise ratio conditions. A nonhomogeneous Poisson-modulated interference process is used to model the ensemble of possible arrival directions of the discrete interferers. Closed-form expressions are derived for the recognition differential of the passive-sonar equation in the presence of Poisson modulated interference. The interference-compensated recognition differential differs from the classical recognition differential by an additive positive term that depend on the interference-to-noise ratio, the directionality of the Poisson modulated interference, and the array beam pattern. PMID- 11051503 TI - Turboprop and rotary-wing aircraft flight parameter estimation using both narrow band and broadband passive acoustic signal-processing methods. AB - Flight parameter estimation methods for an airborne acoustic source can be divided into two categories, depending on whether the narrow-band lines or the broadband component of the received signal spectrum is processed to estimate the flight parameters. This paper provides a common framework for the formulation and test of two flight parameter estimation methods: one narrow band, the other broadband. The performances of the two methods are evaluated by applying them to the same acoustic data set, which is recorded by a planar array of passive acoustic sensors during multiple transits of a turboprop fixed-wing aircraft and two types of rotary-wing aircraft. The narrow-band method, which is based on a kinematic model that assumes the source travels in a straight line at constant speed and altitude, requires time-frequency analysis of the acoustic signal received by a single sensor during each aircraft transit. The broadband method is based on the same kinematic model, but requires observing the temporal variation of the differential time of arrival of the acoustic signal at each pair of sensors that comprises the planar array. Generalized cross correlation of each pair of sensor outputs using a cross-spectral phase transform prefilter provides instantaneous estimates of the differential times of arrival of the signal as the acoustic wavefront traverses the array. PMID- 11051504 TI - On the relationships between the fixed-f1, fixed-f2, and fixed-ratio phase derivatives of the 2f1-f2 distortion product otoacoustic emission. AB - For primary frequency ratios, f2/f1, in the range 1.1-1.3, the fixed-f1 ("f2 sweep") phase derivative of the 2f1-f2 distortion product otoacoustic emission (DPOAE) is larger than the fixed-f2("f1-sweep") one. It has been proposed by some researchers that part or all of the difference between these delays may be attributed to the so-called cochlear filter "build-up" or response time in the DPOAE generation region around the f2 tonotopic site. The analysis of an approximate theoretical expression for the DPOAE signal [Talmadge et al., J. Acoust. Soc. Am. 104, 1517-1543 (1998)] shows that the contributions to the phase derivatives associated with the cochlear filter response is small. It is also shown that the difference between the phase derivatives can be qualitatively accounted for by assuming the approximate scale invariance of cochlear mechanics. The effects of DPOAE fine structure on the phase derivative are also explored, and it is found that the interpretation of the phase derivative in terms of the phase variation of a single DPOAE component can be quite problematic. PMID- 11051505 TI - Nonlinear interactions that could explain distortion product interference response areas. AB - Suppression and/or enhancement of third- and fifth-order distortion products by a third tone that can have a frequency more than an octave above and a level more than 40 dB below the primary tones have recently been measured by Martin et al. [Hear. Res. 136, 105-123 (1999)]. Contours of iso-suppression and iso-enhancement that are plotted as a function of third-tone frequency and level are called interference response areas. After ruling out order aliasing, two possible mechanisms for this effect have been developed, a harmonic mechanism and a catalyst mechanism. The harmonic mechanism produces distortion products by mixing a harmonic of one of the primary tones with the other primary tone. The catalyst mechanism produces distortion products by mixing one or more intermediate distortion products that are produced by the third tone with one or more of the input tones. The harmonic mechanism does not need a third tone and the catalyst mechanism does. Because the basilar membrane frequency response is predicted to affect each of these mechanisms differently, it is concluded that the catalyst mechanism will be dominant in the high-frequency regions of the cochlea and the harmonic mechanism will have significant strength in the low-frequency regions of the cochlea. The mechanisms are dependent on the existence of both even- and odd order distortion, and significant even- and odd-order distortion have been measured in the experimental animals. Furthermore, the nonlinear part of the cochlear mechanical response must be well into saturation when input tones are 50 or more dB SPL. PMID- 11051506 TI - Evidence for spatial tuning in informational masking using the probe-signal method. AB - Auditory spatial attention is one mechanism that may contribute to the ability to identify one sound source in a multi-source environment. The role of auditory spatial attention in a multi-source environment was investigated using the probe signal method. The experiment took place in a quiet room with seven speakers arranged in a semi-circle in front of the listener. The speakers were placed at 30-degree intervals at a distance of 5 ft from the listener. The signal was comprised of eight contiguous, 60-ms pure-tone bursts arranged in either a rising or falling frequency pattern. Masker components were also comprised of eight contiguous pure-tone bursts but with durations that varied randomly from 20 to 100 ms. The six maskers were played with the signal and were constructed in order to result in informational rather than energetic masking. The frequency of each masker component was chosen randomly on each burst from a narrow frequency band, independent from the signal frequency band. The task was 1I-2AFC fixed-level identification with response time measurement. The listener was instructed to focus attention on a specified speaker (expected location) for a block of trials. Accuracy and response time were compared across two conditions: (1) signal presented at the expected location and (2) signal presented at an unexpected location. Results indicate a significant increase in accuracy and faster response time when the signal was presented at the expected location as compared to an unexpected location. These results suggest that auditory spatial attention plays an important role in multi-source listening, especially when the listening environment is complex and uncertain. PMID- 11051507 TI - Effects of ipsilateral and contralateral precursors on overshoot. AB - Overshoot is defined as the decrease in threshold as a brief signal is moved from the beginning to near the temporal center of a longer duration, broadband noise masker. Overshoot can be reduced when another noise (a precursor) is presented just prior to the masker. The purpose of the present investigation was to follow up on a recent psychophysical study which showed that overshoot could be reduced by a precursor presented to the ear contralateral to that receiving the masker and signal. The signal was a 20-ms, 4000-Hz tone that was presented at the beginning or in the temporal center of a 400-ms broadband noise masker. In the first experiment, a 200-ms broadband precursor was presented either to the ipsilateral or to the contralateral ear. The ipsilateral precursor reduced overshoot for all ten subjects, but the contralateral precursor reduced overshoot for only four of the ten subjects. In a supplementary experiment, the contralateral precursor failed to reduce overshoot in a new group of five subjects, both when tested with supra-aural headphones and with insert earphones. In the second experiment, the four subjects who showed an effect of the contralateral precursor in experiment 1 were tested under conditions where the bandwidth of the precursor was manipulated, resulting in either a narrow-band precursor centered at 4000 Hz, a low-band precursor with energy primarily below 4000 Hz, or a high-band precursor with energy primarily above 4000 Hz. There was a tendency for the effectiveness of the ipsilateral and contralateral precursors to be affected similarly (though to different degrees) by changes in the spectral content of the precursor. These results suggest that the effect of the contralateral precursor is not due to a timing cue, and that the processing underlying the effectiveness of ipsilateral and contralateral precursors may be largely the same. PMID- 11051508 TI - Auditory discrimination in a sound-producing electric fish (Pollimyrus): tone frequency and click-rate difference detection. AB - Pollimyrus adspersus is a fish that uses simple sounds for communication and has auditory specializations for sound-pressure detection. The sounds are species specific, and the sounds of individuals are sufficiently stereotyped that they could mediate individual recognition. Behavioral measurements are presented indicating that Pollimyrus probably can make species and individual discriminations on the basis of acoustic cues. Interclick interval (ICI; 10-40 ms) and frequency (100-1400 Hz) discrimination was assessed using modulations of the fish's electric organ discharge rate in the presence of a target stimulus presented in alternation with an ongoing base stimulus. Tone frequency discrimination was best in the 200-600-Hz range, with the best threshold of 1.7% +/- 0.4% standard error at 500 Hz (or 8.5 Hz +/- 1.9 SE). The just noticeable differences (jnd's) were relatively constant from 100 to 500 Hz (mean 8.7 Hz), then increased at a rate of 13.3 Hz per 100 Hz. For click trains, jnd's increased linearly with ICI. The mean jnd's for 10- and 15-ms ICI were both 300 micros (SE= 0.8 ms at 10-ms ICI, SE= 0.11 ms at 15-ms ICI). The jnd at 20-ms ICI was only 1.1 ms +/- 0.25 SE. PMID- 11051509 TI - Monaural and binaural detection of sinusoidal phase modulation of a 500-Hz tone. AB - The detectability of phase modulation was measured for three subjects in two alternative temporal forced-choice experiments. In experiment 1, the detectability of sinusoidal phase modulation in a 1500-ms burst of an 80-dB (SPL), 500-Hz sinusoidal carrier presented to the left ear (monaural condition) was measured. The experiment was repeated with an 80-dB, 500-Hz static (unmodulated) tone at the right ear (dichotic condition). At a modulation rate of 1 Hz, subjects were an order of magnitude more sensitive to phase modulation in the dichotic condition than in the monaural condition. The dichotic advantage decreased monotonically with increasing modulation rate. Subjects ceased to detect movement in the dichotic stimulus above 10 Hz, but a dichotic advantage remained up to a modulation rate of 40 Hz. Thus, although sound movement detection is sluggish, detection of internal phase modulation is not. In experiment 2, thresholds for detecting 2-Hz phase modulation were measured in the dichotic condition as a function of the level of the pure tone in the right ear. The dichotic advantage persisted even when the level of the pure tone was reduced by 50 dB or more. The findings demonstrate a large dichotic advantage which persists to high modulation rates and which depends very little on interaural level differences. PMID- 11051510 TI - Localization of brief sounds: effects of level and background noise. AB - Listeners show systematic errors in vertical-plane localization of wide-band sounds when tested with brief-duration stimuli at high intensities, but long duration sounds at any comfortable level do not produce such errors. Improvements in high-level sound localization associated with increased stimulus duration might result from temporal integration or from adaptation that might allow reliable processing of later portions of the stimulus. Free-field localization judgments were obtained for clicks and for 3- and 100-ms noise bursts presented at sensation levels from 30 to 55 dB. For the brief (clicks and 3-ms) stimuli, listeners showed compression of elevation judgments and increased rates and unusual patterns of front/back confusion at sensation levels higher than 40-45 dB. At lower sensation levels, brief sounds were localized accurately. The localization task was repeated using 3-ms noise burst targets in a background of spatially diffuse, wide-band noise intended to pre-adapt the system prior to the target onset. For high-level targets, the addition of background noise afforded mild release from the elevation compression effect. Finally, a train of identical, high-level, 3-ms bursts was found to be localized more accurately than a single burst. These results support the adaptation hypothesis. PMID- 11051511 TI - The meaning of the Kelly-Lochbaum acoustic-tube model. AB - The scattering equations of the Kelly-Lochbaum segmented tube, including the time varying extension by Strube, are originally based on the assumption of uniform spatial segments and stepwise time update of the acoustic impedances. Here, it is shown that the same equations can be derived without these assumptions for a nonuniform time-varying tube from the discretization of space and time derivatives by the bilinear z transform or by centered differences along the rotated coordinates ct+/-x. Moreover, the same equations also hold for a chain of lattice circuits (or equivalents) with appropriate parameters, if time derivatives are discretized by the bilinear z transform. These circuits can also be extended to simulate uniform segments of varying length. PMID- 11051512 TI - Time-varying spectral change in the vowels of children and adults. AB - Recent studies have shown that time-varying changes in formant pattern contribute to the phonetic specification of vowels. This variation could be especially important in children's vowels, because children have higher fundamental frequencies (f0's) than adults, and formant-frequency estimation is generally less reliable when f0 is high. To investigate the contribution of time-varying changes in formant pattern to the identification of children's vowels, three experiments were carried out with natural and synthesized versions of 12 American English vowels spoken by children (ages 7, 5, and 3 years) as well as adult males and females. Experiment 1 showed that (i) vowels generated with a cascade formant synthesizer (with hand-tracked formants) were less accurately identified than natural versions; and (ii) vowels synthesized with steady-state formant frequencies were harder to identify than those which preserved the natural variation in formant pattern over time. The decline in intelligibility was similar across talker groups, and there was no evidence that formant movement plays a greater role in children's vowels compared to adults. Experiment 2 replicated these findings using a semi-automatic formant-tracking algorithm. Experiment 3 showed that the effects of formant movement were the same for vowels synthesized with noise excitation (as in whispered speech) and pulsed excitation (as in voiced speech), although, on average, the whispered vowels were less accurately identified than their voiced counterparts. Taken together, the results indicate that the cues provided by changes in the formant frequencies over time contribute materially to the intelligibility of vowels produced by children and adults, but these time-varying formant frequency cues do not interact with properties of the voicing source. PMID- 11051513 TI - Sources of listener disagreement in voice quality assessment. AB - Traditional interval or ordinal rating scale protocols appear to be poorly suited to measuring vocal quality. To investigate why this might be so, listeners were asked to classify pathological voices as having or not having different voice qualities. It was reasoned that this simple task would allow listeners to focus on the kind of quality a voice had, rather than how much of a quality it possessed, and thus might provide evidence for the validity of traditional vocal qualities. In experiment 1, listeners judged whether natural pathological voice samples were or were not primarily breathy and rough. Listener agreement in both tasks was above chance, but listeners agreed poorly that individual voices belonged in particular perceptual classes. To determine whether these results reflect listeners' difficulty agreeing about single perceptual attributes of complex stimuli, listeners in experiment 2 classified natural pathological voices and synthetic stimuli (varying in f0 only) as low pitched or not low pitched. If disagreements derive from difficulties dividing an auditory continuum consistently, then patterns of agreement should be similar for both kinds of stimuli. In fact, listener agreement was significantly better for the synthetic stimuli than for the natural voices. Difficulty isolating single perceptual dimensions of complex stimuli thus appears to be one reason why traditional unidimensional rating protocols are unsuited to measuring pathologic voice quality. Listeners did agree that a few aphonic voices were breathy, and that a few voices with prominent vocal fry and/or interharmonics were rough. These few cases of agreement may have occurred because the acoustic characteristics of the voices in question corresponded to the limiting case of the quality being judged. Values of f0 that generated listener agreement in experiment 2 were more extreme for natural than for synthetic stimuli, consistent with this interpretation. PMID- 11051514 TI - Effects of the salience of pitch and periodicity information on the intelligibility of four-channel vocoded speech: implications for cochlear implants. AB - Recent simulations of continuous interleaved sampling (CIS) cochlear implant speech processors have used acoustic stimulation that provides only weak cues to pitch, periodicity, and aperiodicity, although these are regarded as important perceptual factors of speech. Four-channel vocoders simulating CIS processors have been constructed, in which the salience of speech-derived periodicity and pitch information was manipulated. The highest salience of pitch and periodicity was provided by an explicit encoding, using a pulse carrier following fundamental frequency for voiced speech, and a noise carrier during voiceless speech. Other processors included noise-excited vocoders with envelope cutoff frequencies of 32 and 400 Hz. The use of a pulse carrier following fundamental frequency gave substantially higher performance in identification of frequency glides than did vocoders using envelope-modulated noise carriers. The perception of consonant voicing information was improved by processors that preserved periodicity, and connected discourse tracking rates were slightly faster with noise carriers modulated by envelopes with a cutoff frequency of 400 Hz compared to 32 Hz. However, consonant and vowel identification, sentence intelligibility, and connected discourse tracking rates were generally similar through all of the processors. For these speech tasks, pitch and periodicity beyond the weak information available from 400 Hz envelope-modulated noise did not contribute substantially to performance. PMID- 11051515 TI - Localization of multiple sound sources with two microphones. AB - This paper presents a two-microphone technique for localization of multiple sound sources. Its fundamental structure is adopted from a binaural signal-processing scheme employed in biological systems for the localization of sources using interaural time differences (ITD). The two input signals are transformed to the frequency domain and analyzed for coincidences along left/right-channel delay line pairs. The coincidence information is enhanced by a nonlinear operation followed by a temporal integration. The azimuths of the sound sources are estimated by integrating the coincidence locations across the broadband of frequencies in speech signals (the "direct" method). Further improvement is achieved by using a novel "stencil" filter pattern recognition procedure. This includes coincidences due to phase delays of greater than 2pi, which are generally regarded as ambiguous information. It is demonstrated that the stencil method can greatly enhance localization of lateral sources over the direct method. Also discussed and analyzed are two limitations involved in both methods, namely missed and artifactual sound sources. Anechoic chamber tests as well as computer simulation experiments showed that the signal-processing system generally worked well in detecting the spatial azimuths of four or six simultaneously competing sound sources. PMID- 11051516 TI - Chinese dialect identification using segmental and prosodic features. AB - Several approaches to Chinese dialect identification based on segmental and prosodic features of speech are described in this paper. When using segmental information only, the system performs phonotactic analysis after speech utterances have been tokenized into sequences of broad phonetic classes. The second scheme comprises prosodic models which are trained to capture tone sequence information for individual dialects. Also proposed is a novel approach that examines differences between Chinese dialects at broad phonetic and prosodic levels. These algorithms were evaluated via a multispeaker read-speech mode. Simulation results indicate that the combined use of segmental and prosodic features allows the proposed system to discriminate among three major Chinese dialects spoken in Taiwan with 93.0% accuracy. PMID- 11051517 TI - Angular scatter ultrasound imaging of wavelength scale targets. AB - A bistatic ultrasound imaging system is demonstrated that uses two 32-element linear phased array transducers oriented at an angle of 40 degrees to one another. The system simultaneously acquires and displays in real time one conventional backscatter image and one "angular scatter" image formed using side scattered echoes from the same B-mode sector region. Experiments are presented that show differences in the magnitudes of backscatter and angular scatter signals acquired from three nylon monofilaments with diameters less than one wavelength and from soft tissue structures in vivo. The relative magnitudes of angular scatter signals from the monofilaments are qualitatively consistent with a theoretical analysis of acoustic scattering from elastic cylinders. Larger tissue features are more clearly defined in angular scatter images. This result is attributed to the orientation of specularly reflecting surfaces and the expected influence of scattering angle on the system's sensitivity to different scatterer spacings. PMID- 11051518 TI - Analysis of acoustic communication by ants. AB - An analysis is presented of acoustic communication by ants, based on near-field theory and on data obtained from the black imported fire ant Solenopsis richteri and other sources. Generally ant stridulatory sounds are barely audible, but they occur continuously in ant colonies. Because ants appear unresponsive to airborne sound, myrmecologists have concluded that stridulatory signals are transmitted through the substrate. However, transmission through the substrate is unlikely, for reasons given in the paper. Apparently ants communicate mainly through the air, and the acoustic receptors are hairlike sensilla on the antennae that respond to particle sound velocity. This may seem inconsistent with the fact that ants are unresponsive to airborne sound (on a scale of meters), but the inconsistency can be resolved if acoustic communication occurs within the near field, on a scale of about 100 mm. In the near field, the particle sound velocity is significantly enhanced and has a steep gradient. These features can be used to exclude extraneous sound, and to determine the direction and distance of a near field source. Additionally, we observed that the tracheal air sacs of S. richteri can expand within the gaster, possibly amplifying the radiation of stridulatory sound. PMID- 11051519 TI - Behavioral responses of humpback whales (Megaptera novaeangliae) to full-scale ATOC signals. AB - Loud (195 dB re 1 microPa at 1 m) 75-Hz signals were broadcast with an ATOC projector to measure ocean temperature. Respiratory and movement behaviors of humpback whales off North Kauai, Hawaii, were examined for potential changes in response to these transmissions and to vessels. Few vessel effects were observed, but there were fewer vessels operating during this study than in previous years. No overt responses to ATOC were observed for received levels of 98-109 dB re 1 microPa. An analysis of covariance, using the no-sound behavioral rate as a covariate to control for interpod variation, found that the distance and time between successive surfacings of humpbacks increased slightly with an increase in estimated received ATOC sound level. These responses are very similar to those observed in response to scaled-amplitude playbacks of ATOC signals [Frankel and Clark, Can. J. Zool. 76, 521-535 (1998)]. These similar results were obtained with different sound projectors, in different years and locations, and at different ranges creating a different sound field. The repeatability of the findings for these two different studies indicates that these effects, while small, are robust. This suggests that at least for the ATOC signal, the received sound level is a good predictor of response. PMID- 11051520 TI - Sounds produced by Australian Irrawaddy dolphins, Orcaella brevirostris. AB - Sounds produced by Irrawaddy dolphins, Orcaella brevirostris, were recorded in coastal waters off northern Australia. They exhibit a varied repertoire, consisting of broadband clicks, pulsed sounds and whistles. Broad-band clicks, "creaks" and "buzz" sounds were recorded during foraging, while "squeaks" were recorded only during socializing. Both whistle types were recorded during foraging and socializing. The sounds produced by Irrawaddy dolphins do not resemble those of their nearest taxonomic relative, the killer whale, Orcinus orca. Pulsed sounds appear to resemble those produced by Sotalia and nonwhistling delphinids (e.g., Cephalorhynchus spp.). Irrawaddy dolphins exhibit a vocal repertoire that could reflect the acoustic specialization of this species to its environment. PMID- 11051521 TI - Scattering of ultrasonic waves by void inclusions. AB - The Local Interaction Simulation Approach (LISA) has been applied to study the propagation of ultrasonic pulses in materials containing void inclusions. First the case of a single scatterer has been considered, with the conclusion that a void must be represented by at least four cells of the discretization lattice. Then the case of ultrasound propagation in a material with both a regular and random distribution of inclusions has been studied. Interesting interference effects have been obtained and it can be concluded that in both cases LISA allows reliable and efficient simulations to be performed. PMID- 11051522 TI - Optical measurement of the speed of sound in air over the temperature range 300 650 K. AB - Using laser-induced thermal acoustics (LITA), the speed of sound in room air (1 atm) is measured over the temperature range 300-650 K. Since the LITA apparatus maintains a fixed sound wavelength as temperature is varied, this temperature range simultaneously corresponds to a sound frequency range of 10-15 MHz. The data are compared to a published model and typically agree within 0.1%-0.4% at each of 21 temperatures. PMID- 11051523 TI - Phase and group velocities of fast and slow compressional waves in trabecular bone. AB - This Letter is an extension to a multilayer model of porous bone first proposed by Hughes et al. [Ultrasound Med. Biol. 25, 811-821 (1999)]. Both slow and fast compressional waves propagate when the acoustic wave propagation is parallel to the trabecular alignment. However, a slow wave disappears at high refraction angles. To explain this phenomenon, the multilayer model is extended to compute group velocity surface and arrival times with an angle. Two major effects are highlighted as the refraction angle increases. First, the energy of the slow wave is refracted from the phase propagation direction. Second, the signals of fast and slow waves overlap. As a consequence, the slow wave may not be observed for a refraction angle greater than 40 degrees, which is in agreement with previous experimental data published by Hughes et al. and others. PMID- 11051524 TI - Measurement of prolonged fatigue in the working population: determination of a cutoff point for the checklist individual strength. AB - In the Netherlands, a large-scale prospective cohort study was started on prolonged fatigue in the working population. The 1st issue that had to be addressed was the determination of a cutoff point for fatigue for use in the working population. Fatigue is measured with the Checklist Individual Strength (CIS), a 20-item self-report questionnaire. This article demonstrates the process of decision making in the determination of the cutoff point. Total CIS scores were calculated, sensitivity and specificity were compared for potential cutoff points, and a receiver operating characteristics analysis was conducted. A CIS total cutoff point for fatigue of >76 was determined, with a specificity of 90% and a sensitivity of 73%. Limitations regarding the use of cutoff points are discussed. It is concluded that the defined cutoff point seems to be appropriate for use in the working population. PMID- 11051525 TI - Linking manager values and behavior with employee values and behavior: a study of values and safety in the hairdressing industry. AB - Five theoretical processes that link values and behavior were identified: value congruence, value-behavior consistency, behavioral modeling, value internalization, and descriptive norms. A values questionnaire was administered to 219 employees and their managers. Values for preventive safety procedures and time urgency were linked to safety behavior of employees in the hairdressing industry. Hairdressers are frequently exposed to hazardous chemicals, and the safety behavior measured was wearing protective gloves. Results support value internalization (linking manager's and employee's values) and behavioral modeling (linking manager's and employee's behavior). Employee time urgency values were also negatively related to safety behavior (value-behavior consistency). Descriptive norms and value congruence were not supported. Strategies to align values within organizations and the management of safety at work are considered. PMID- 11051526 TI - Gender differences in job strain, social support at work, and psychological distress. AB - Using the demand-control-support model of job strain, the authors examined gender differences in the relationship between psychosocial work exposures and psychological distress in a cross-sectional sample of 7,484 employed Canadians. Compared with low-strain work, high-strain and active work were associated with a significantly higher level of distress in both men and women. Differences in psychological distress in relation to psychosocial work exposures were greater for men than for women. Low social support was associated with higher distress across all categories of job strain, and the combined effect of low social support and high job strain was associated with the greatest increase in distress. This pattern was similar in men and women. This study suggests that psychosocial work exposures may be a more significant determinant of psychological well-being in male workers compared with female workers. PMID- 11051527 TI - Disruptions to women's social identity: a comparative study of workplace stress experienced by women in three geographic regions. AB - Drawing on social identity theory (P. J. Burke, 1991) and the current status of women and equal opportunity legislation, the authors tested several factors associated with distress in working women in the People's Republic of China (PRC), Hong Kong, and the United States. Women in Hong Kong experienced significantly greater levels of life stress than PRC and U.S. women. Reports of negative attitudes toward women, gender evaluation, and avoidance coping were greater for Hong Kong and PRC women than for U.S. women. Hong Kong women reported more use of positive/confrontational coping mechanisms. Negative attitudes toward women had an important influence on life stress across regions. Moderator tests resulted in 2 significant findings: The effect of negative attitudes toward women on life stress was stronger for PRC and Hong Kong women, and the relationship between nervous/self-destructive coping and life stress was stronger for U.S. women. PMID- 11051528 TI - Employee retaliation: the neglected consequence of poor leader-member exchange relations. AB - Although the beneficial effects of high-quality leader-member exchange (LMX) relationships have been well-documented in the leadership literature, much less is known about the potentially damaging effects of poor exchange relationships. Using 150 intact leader-member dyads, the authors investigated the relationship between LMX and supervisors' reports of employee retaliation behavior, performance, and citizenship. Results indicated that performance and citizenship were positively related to LMX. More important, LMX was negatively correlated with retaliation behavior. Supervisors reported that subordinates in poor exchange relationships were more likely to engage in retaliation against the organization than subordinates in high-quality relationships. The lack of a high quality exchange relationship was, therefore, not just associated with the absence of positive consequences but also led to reports of potentially disruptive behaviors. PMID- 11051529 TI - A comparison of the stress-strain process for business owners and nonowners: differences in job demands, emotional exhaustion, satisfaction, and social support. AB - One hundred sixty licensed morticians were surveyed to examine differences among business owners, managers, and employees on the relations proposed by G. F. Koeske and R. D. Koeske's (1993) stressor-strain-outcome model. Forty-eight percent of the morticians were owners, 16% were managers, and 36% were employees. Owners had less social support from work-related sources and perceived lower levels of role ambiguity and role conflict, less emotional exhaustion, and higher levels of job satisfaction and professional satisfaction than did nonowners. Social support from work-related sources and ownership each moderated the relationship between emotional exhaustion and job satisfaction but not between emotional exhaustion and professional satisfaction. Emotional exhaustion partially mediated the effect of stressors on job satisfaction and professional satisfaction. PMID- 11051530 TI - Prediction of hourly microenvironmental concentrations of fine particles based on measurements obtained from the Baltimore scripted activity study. AB - Researchers have developed a variety of computer-based models to estimate population exposure to air pollution. These models typically estimate exposures by simulating the movement of specific population groups through defined microenvironments. During the summer of 1998 and winter of 1999, researchers with the Harvard School of Public Health (HSPH) conducted a field study in Baltimore, MD, to acquire data for improving microenvironmental models. Using a special roll around instrument system, a technician measured 1- and 12-h pollutant concentrations while engaging in scripted sequences of activities typical of retirees. Each scripted activity assigned the technician to a geographic location and to a microenvironment. The technician recorded special conditions associated with each activity (e.g., open windows, environmental tobacco smoke) in a real time diary. Data on ambient pollutant levels, temperature, and other potential explanatory factors were also collected. Eleven pollutants were measured by the roll-around instrument system, including particulate matter with an aerodynamic diameter less than 2.5 microm (PM2.5), ozone, carbon monoxide, and benzene. This article presents the results of statistical analyses performed solely on the 1-h PM2.5 data measured by a DustTrak monitor, which ranged from 1.5 to 444.8 microg/m3 with a median value of 14.6 microg/m3. Results of stepwise linear regression (SLR) suggest that PM2.5 exposure is significantly increased by passive smoking, high ambient PM2.5 concentrations reported by fixed-site monitors, food preparation, charcoal grills, car travel, outdoor roadside locations, and high humidity. Analysts should explicitly represent the effects of these parameters within any model developed to estimate population exposure to PM2.5. In a related study, a panel of volunteer retirees each carried a personal PM2.5 monitor and a real-time diary for nominal 24-h sampling periods as they engaged in normal daily activities. A regression equation derived from SLR analysis of the scripted activity database was applied to eight subject-days of diary data provided by the volunteer seniors to produce estimates of PM2.5 exposure for each event documented in each diary. The event-specific exposure estimates were then averaged over all events in each sampling period to produce nominal 24-h average exposure estimates. The absolute difference between the estimate obtained from the regression equation and the corresponding personal monitor measurement averaged 13%. The fixed-site monitors generally provided poorer estimates of exposure; the absolute differences for the Old Town and Clifton Park monitors averaged 26.7% and 19.5%, respectively, of the personal monitor values. PMID- 11051531 TI - Coarse and fine particles and daily mortality in the Coachella Valley, California: a follow-up study. AB - Many epidemiological studies provide evidence of an association between ambient particles, measured as PM10, and daily mortality. Most of these studies have been conducted in urban areas where PM10 is highly correlated with and dominated by fine particles less than 2.5 microm in diameter (PM2.5). Fewer studies have investigated impacts associated with the fraction of coarse mode particles (between 2.5 and 10 microm in diameter). In a previous study using data from 1989 through 1992 in the Coachella Valley, a desert resort and retirement area east of Los Angeles, we reported associations between PM10 and several different measures of mortality [Ostro B.D., Hurley S., and Lipsett M.J. Air pollution and daily mortality in the Coachella Valley, California: a study of PM10 dominated by coarse particles. Environ. Res. 1999: 81: 231-238]. In this arid environment, coarse particles of geologic origin are highly correlated with and comprise approximately 60% of PM10, increasing to >90% during wind events. This study was intended to repeat the earlier investigation using 10 years (1989-1998) of daily data on mortality and PM10. The last 2.5 years of data also included daily measures of PM2.5, allowing examination of size-specific impacts. To ensure adequate statistical power, we attempted to develop predictive models for both fine and coarse particles to use in analyses of the full 10-year period. An acceptable fit was found only for coarse particles, which were found to be a cubic function of PM10 (R2 = 0.95). Outcome variables included several measures of daily mortality, including all-cause (minus accidents and homicides), cardiovascular and respiratory mortality. Multivariate Poisson regression analyses using generalized additive models were employed to explain the variation in these endpoints, controlling for temperature, humidity, day of the week, season, and time, using locally weighted smoothing techniques. Pollution lags of up to 4 days were examined. Several pollutants were associated with all-cause mortality, including PM2.5, carbon monoxide and nitrogen dioxide. More consistent results were found for cardiovascular-specific mortality, for which associations were found for coarse particles (RR = 1.02; 95% C.I., 1.01-1.04), PM10 (RR = 1.03; 95% C.I., 1.01-1.05). None of the pollutants was associated with respiratory-specific mortality. Ozone was not associated with any of the mortality outcomes. These findings are generally consistent with those we previously reported for the Coachella Valley for the period 1989-1992, demonstrating associations between several measures of particulate matter and daily mortality in an environment in which particulate concentrations are dominated by the coarse fraction. PMID- 11051532 TI - Methodological approaches to the analysis of hierarchical studies of air pollution and respiratory health--examples from the CESAR study. Central European Study on Air pollution and Respiratory Health. AB - OBJECTIVES: Many studies of air pollution and health are carried out over several geographical areas, and sometimes over several countries. This paper explores three approaches to analysis in such studies: a non hierarchical model, a two stage analysis, and multilevel modelling. Illustrations are given using a preliminary subset of data from the CESAR study. DESIGN: The Central European Study on Air pollution and Respiratory Health (CESAR) was conducted in 25 areas within six Central European countries, enrolling 20,271 schoolchildren. Pollution averages were calculated for each area. Associations between pollution and health outcomes were estimated under different models. MAIN RESULTS: A regression analysis of log FVC (forced vital capacity) on PM10, ignoring the geographical hierarchy, estimated a significant mean drop in FVC (adjusted for confounders) of 2.2% (95% CI 0.5% to 1.3%), p=0.007, from the area with the lowest PM10 to that with the highest. A multilevel model (mlm), using data for all children, but with random effects at area and country level, estimated a drop of 2.8% (-0.6% to 6.1%), p=0.110. A two-stage analysis (mean log FVC, adjusted for confounders, was estimated for each area using regression, and these means then regressed on PM10) estimated a drop of 2.6% (-0.5% to 5.5%), p=0.101. Simulation exercises showed the non hierarchical method to be very inadequate in the context of the CESAR study, with only half of all 95% confidence intervals for the estimated PM10 slope containing the true value (i.e., that used to create the simulated data). The two-stage and multilevel modelling methods gave results which were substantially better, though both underperformed slightly. All three methods appeared to give unbiased slope estimates. CONCLUSIONS: Acknowledgement of hierarchical structures is essential in statistical inference--standard errors can be substantially incorrect when they are ignored. Multilevel, random-effects models correctly address hierarchical structures, though having few units at higher levels can cause problems in convergence, especially where complex modelling is required. Two-stage analyses, acknowledging hierarchy, provide simple alternatives to random-effects models. PMID- 11051533 TI - Relationships of mortality with the fine and coarse fractions of long-term ambient PM10 concentrations in nonsmokers. AB - In a cohort of 6338 California Seventh-day Adventists, we previously observed for males associations between long-term concentrations of particulate matter (PM) with an aerodynamic diameter less than 10 microm (PM10) and 15-year mortality due to all natural causes (ANC) and lung cancer (LC) listed as underlying causes of death and due to nonmalignant respiratory disease listed as either the underlying or a contributing (CRC) cause of death. The purpose of this analysis was to determine whether these outcomes were more strongly associated with the fine (PM2.5) or the coarse (PM2.5-10) fractions of PM10. For participants who lived near an airport (n=3769), daily PM2.5 concentrations were estimated from airport visibility, and on a monthly basis, PM2.5-10 concentrations were calculated as the differences between PM10 and PM2.5. Associations between ANC, CRC, and LC mortality (1977-1992) and mean PM10, PM2.5, and PM2.5-10 concentrations at study baseline (1973-1977) were assessed using Cox proportional hazards models. Magnitudes of the PM10 associations for the males of this subgroup were similar to those for the males in the entire cohort although not statistically significant due to the smaller numbers. In single-pollutant models, for an interquartile range (IQR) increase in PM10 (29.5 microg/m3), the rate ratios (RRs) and 95% confidence intervals (CI) were 1.15 (0.94, 1.41) for ANC, 1.48 (0.93, 2.34) for CRC, and 1.84 (0.59, 5.67) for LC. For an IQR increase in PM2.5 (24.3 microg/m3), corresponding RRs (95% CI) were 1.22 (0.95, 1.58), 1.64 (0.93, 2.90), and 2.23 (0.56, 8.94), and for an IQR increase in PM2.5-10 (9.7 microg/m3), corresponding RRs (95% CI) were 1.05 (0.92, 1.20), 1.19 (0.88, 1.62), and 1.25 (0.63, 2.49), respectively. When both PM25 and PM2.5-10 were entered into the same model, the PM2.5 estimates remained stable while those of PM2.5-10 decreased. We concluded that previously observed associations of long-term ambient PM10 concentration with mortality for males were best explained by a relationship of mortality with the fine fraction of PM10 rather than with the coarse fraction of PM10. PMID- 11051534 TI - Evaluation of a real-time passive personal particle monitor in fixed site residential indoor and ambient measurements. AB - Recent experimental findings in animals and humans indicate adverse respiratory effects from short-term exposures to particulate air pollutants, especially in sensitive subpopulations such as asthmatics. The relationship between air pollution and asthma has mainly been determined using particulate matter (PM) measurements from central sites. Validated tools are needed to assess exposures most relevant to health effects. Recently, a personal passive particulate sampler (personal Data-RAM, pDR, MIE Inc., Bedford, MA) has become available for studying personal exposures to PM with time resolution at 1 min. The pDR measures light scatter from PM in the 0.1-10 microM range, the significant range for health effects. In order to assess the ability of the pDR in predicting gravimetric mass, pDRs were collocated with PM2.5 and PM10 Harvard Impactors (HI) inside and outside nine homes of asthmatic children and at an outdoor central Air Pollution Control District site. Results are presented of comparisons between the HI samplers and the pDR in various modes of operation: passive, active, and active with a heated inlet. When used outdoors at fixed sites the pDR readings exhibit interference from high relative humidity (RH) unless operated with a method for drying inlet air such as a heater, or if readings at times of high RH are adjusted. The pDR correlates more highly with the HI PM2.5 than with the HI PM10 (r2 = 0.66 vs. 0.13 for outdoors, r2 = 0.42 vs. 0.20 for indoors). The pDR appears to be a useful tool for an epidemiologic study that aims to examine the relationship between health outcomes and personal exposure to peaks in PM. PMID- 11051535 TI - Interim results of the study of particulates and health in Atlanta (SOPHIA). AB - Substantial evidence supports an association of particulate matter (PM) with cardiorespiratory illnesses, but little is known regarding characteristics of PM that might contribute to this association and the mechanisms of action. The Atlanta superstation sponsored by the Electric Power Research Institute as part of the Aerosol Research and Inhalation Epidemiology Study (ARIES) study is monitoring chemical composition of ambient particles by size fraction, as well as a comprehensive suite of other pollutants, at a site in downtown Atlanta during the 25-month period, August 1, 1998-August 31, 2000. Our investigative team is making use of this unique resource in several morbidity studies, called the "Study of Particulates and Health in Atlanta (SOPHIA)". The study includes the following components: (1) a time series investigation of emergency department (ED) visits for the period during which the superstation is operating; (2) a time series investigation of ED visits during the 5 years prior to implementation of the superstation; and (3) a study of arrhythmic events in patients equipped with automatic implantable cardioverter defibrillators (AICDs) for the period January 1, 1993-August 31, 2000. Thirty-three of 39 Atlanta area EDs are participating in the ED studies, comprising over a million annual ED visits. In this paper, we present initial analyses of data from 18 of the 33 participating EDs. The preliminary data set includes 1,662,713 ED visits during the pre-superstation time period and 559,480 visits during the superstation time period. Visits for four case groupings--asthma, chronic obstructive pulmonary disease (COPD), dysrhythmia, and all cardiovascular diseases (CVDs) combined--have been assessed relative to daily air quality indices, controlling for long-term temporal trends and meteorologic variables, using general linear models, generalized estimating equations and generalized additive models. Single-pollutant models predicting case visitation rates using moving averages of 0-, 1-, and 2-day lagged air quality variables were run. For the pre-superstation period, PM10 (24-h), ozone (8-h), SO2 (1-h), NO2 (1-h) and CO (1-h) were studied. For the first 12 months of superstation operation, the following air quality variables of a priori interest were available: ozone (8-h), NO2 (1-h), SO2 (1-h), CO (1-h), and 24-h measurements of PM10, coarse PM (PM 2.5-10 microm), PM2.5, polar VOCs, 10-100 nm particulate count and surface area, and in the PM2.5 fraction: sulfates, acidity, water-soluble metals, organic matter (OM), and elemental carbon (EC). During the pre-superstation time period, statistically significant, positive associations were observed for adult asthma with ozone, and for COPD with ozone, NO2 and PM10. During the superstation time period, the following statistically significant, positive associations were observed: dysrhythmia with CO, coarse PM, and PM2.5 EC; and all CVDs with CO, PM2.5 EC and PM2.5 OM. While covariation of many of the air quality indices limits the informativeness of this analysis, the study provides one of the first assessments of PM components in relation to ED visits. PMID- 11051536 TI - Air pollution, aeroallergens and cardiorespiratory emergency department visits in Saint John, Canada. AB - Existing studies of the association between air pollution, aeroallergens and emergency department (ED) visits have generally examined the effects of a few pollutants or aeroallergens on individual conditions such as asthma or chronic obstructive pulmonary disease. In this study, we considered a wide variety of respiratory and cardiac conditions and an extensive set of pollutants and aeroallergens, and utilized prospectively collected information on possible effect modifiers which would not normally be available from purely administrative data. The association between air pollution, aeroallergens and cardiorespiratory ED visits (n = 19,821) was examined for the period 1992 to 1996 using generalized additive models. ED visit, air pollution and aeroallergen time series were prefiltered using LOESS smoothers to minimize temporal confounding, and a parsimonious model was constructed to control for confounding by weather and day of week. Multipollutant and multi-aeroallergen models were constructed using stepwise procedures and sensitivity analyses were conducted by season, diagnosis, and selected individual characteristics or effect modifiers. In single-pollutant models, positive effects of all pollutants but NO2 and COH were observed on asthma visits, and positive effects on all respiratory diagnosis groups were observed for O3, SO2, PM10, PM2.5, and SO4(2-). Among cardiac conditions, only dysrhythmia visits were positively associated with all measures of particulate matter. In the final year-round multipollutant models, a 20.9% increase in cardiac ED visits was attributed to the combination of O3 (16.0%, 95% CI 2.8 30.9) and SO2 (4.9%, 95%CI 1.7-8.2) at the mean concentration of each pollutant. In the final multipollutant model for respiratory visits, O3 accounted for 3.9% of visits (95% CI 0.8-7.2), and SO2 for 3.7% (95% CI 1.5-6.0), whereas a weak, negative association was observed with NO2. In multi-aeroallergen models of warm season asthma ED visits, Ascomycetes, Alternaria and small round fungal spores accounted for 4.5% (95% CI 1.8-7.4), 4.7% (95% CI 1.0-8.6) and 3.0% (95% CI 0.8 5.1), respectively, of visits at their mean concentrations, and these effects were not sensitive to adjustment for air pollution effects. In conclusion, we observed a significant influence of the air pollution mix on cardiac and respiratory ED visits. Although in single-pollutant models, positive associations were noted between ED visits and some measures of particulate matter, in multipollutant models, pollutant gases, particularly ozone, exhibited more consistent effects. Aeroallergens were also significantly associated with warm season asthma ED visits. PMID- 11051537 TI - Personal exposure to particles in Banska Bystrica, Slovakia. AB - Epidemiological studies have associated adverse health impacts with ambient concentrations of particulate matter (PM), though these studies have been limited in their characterization of personal exposure to PM. An exposure study of healthy nonsmoking adults and children was conducted in Banska Bystrica, Slovakia, to characterize the range of personal exposures to air pollutants and to determine the influence of occupation, season, residence location, and outdoor and indoor concentrations on personal exposures. Twenty-four-hour personal, at home indoor, and ambient measurements of PM10, PM2.5, sulfate (SO4(2-)) and nicotine were obtained for 18 office workers, 16 industrial workers, and 15 high school students in winter and summer. Results showed that outdoor levels of pollutants were modest, with clear seasonal differences: outdoor PM10 summer/winter mean = 35/45 microg/m3; PM2.5 summer/winter mean = 22/32 microg/m3. SO4(2-) levels were low (4-7 microg/m3) and relatively uniform across the different sample types (personal, indoor, outdoor), areas, and occupational groups. This suggests that SO4(2-) may be a useful marker for combustion mode particles of ambient origin, although the relationship between personal exposures and ambient SO4(2-) levels was more complex than observed in North American settings. During winter especially, the central city area showed higher concentrations than the suburban location for outdoor, personal, and indoor measures of PM10, PM2.5, and to a lesser extent for SO4(2-), suggesting the importance of local sources. For PM2.5 and PM10, ratios consistent with expectations were found among exposure indices for all three subject groups (personal>indoor>outdoor), and between work type (industrial>students>office workers). The ratio of PM2.5 personal to indoor exposures ranged from 1.0 to 3.9 and of personal to outdoor exposures from 1.6 to 4.2. The ratio of PM10 personal to indoor exposures ranged from 1.1 to 2.9 and the ratio of personal to outdoor exposures from 2.1 to 4.1. For a combined group of office workers and students, personal PM10/PM2.5 levels were predicted by statistically significant multivariate models incorporating indoor (for PM2.5) or outdoor (for PM10) PM levels, and nicotine exposure (for PM10). Small but significant fractions of the overall variability, 15% for PM2.5 and 17% for PM10, were explained by these models. The results indicate that central site monitors underpredict actual human exposures to PM2.5 and PM10. Personal exposure to SO4(2-) was found to be predicted by outdoor or indoor SO4(2-) levels with 23-71% of the overall variability explained by these predictors. We conclude that personal exposure measurements and additional demographic and daily activity data are crucial for accurate evaluation of exposure to particles in this setting. PMID- 11051538 TI - Effects of ambient particle pollution on daily mortality in Melbourne, 1991-1996. AB - This paper presents the results of a study in Melbourne, Australia, of the short term effects of ambient fine particle pollution on daily mortality occurring between 1991 and 1996. The methodological approach used Poisson regression and Generalized Additive Models (GAM) with LOESS smoothing to control for temporal and meteorological effects. The association between particles and increases in daily mortality was examined using nephelometry data (bsp, mean 24 h average = 0.26 x 10(-4) m(-1), mean 1 h maximum = 0.60 x 10(-4) m(-1)), PM2.5 (24 h mean = 9.42 microg/m3) and PM10 (24 h mean = 19 microg/m3). Both the PM10 and PM2.5 data were estimated from nephelometry data using previously derived relationships for the Melbourne airshed. Significant positive associations between the particle measures considered and all cause and respiratory mortality were found in the warm season (November-March). A 1 x 10(-4) m(-1) increase in maximum 1-h bsp levels was associated with a 2.19% (95% CI = 0.01-4.43%) increase in risk of death for all cause mortality and a 10.40% (95% CI = 2.44-18.97%) increase in risk for respiratory mortality in the warm season. A 1 microg/m3 increase in 24-h PM2.5 in the warm season was associated with a 0.38% (95% CI = 0.06-0.70%) increase in risk of death for all cause mortality and a 1.18% (95% CI = 0.05 2.32%) increase in risk for respiratory mortality. For PM10, a 1 microg/m3 increase was associated with an increased risk of 0.18% (95% CI = 0.03-0.33%) for all cause mortality and 0.59% (95% CI = 0.06-1.13%) for respiratory mortality. Significant associations were also found in the 65+ age group in the warm season. However, for these warm periods, the effects of ozone (average 1 h maximum = 127 ppb) and nitrogen dioxide (average 1 h maximum = 70.7 ppb) were also significant and, due to high correlations between these pollutants, it was not possible to separate the particle effects from those of O3 and NO2. Sulfur dioxide was not examined as concentrations of this pollutant in Melbourne are very low (max 1 h = 15-24 ppb, annual average 0.8 ppb). Comparison with other Australian studies in Sydney and Brisbane indicates different results for particle pollution. PMID- 11051539 TI - Comparison of PM2.5 and PM10 monitors. AB - An extensive PM monitoring study was conducted during the 1998 Baltimore PM Epidemiology-Exposure Study of the Elderly. One goal was to investigate the mass concentration comparability between various monitoring instrumentation located across residential indoor, residential outdoor, and ambient sites. Filter-based (24-h integrated) samplers included Federal Reference Method Monitors (PM2.5 FRMs), Personal Environmental Monitors (PEMs), Versatile Air Pollution Samplers (VAPS), and cyclone-based instruments. Tapered element oscillating microbalances (TEOMs) collected real-time data. Measurements were collected on a near-daily basis over a 28-day period during July-August, 1998. The selected monitors had individual sampling completeness percentages ranging from 64% to 100%. Quantitation limits varied from 0.2 to 5.0 microg/m3. Results from matched days indicated that mean individual PM10 and PM2.5 mass concentrations differed by less than 3 microg/m3 across the instrumentation and within each respective size fraction. PM10 and PM2.5 mass concentration regression coefficients of determination between the monitors often exceeded 0.90 with coarse (PM10-2.5) comparisons revealing coefficients typically well below 0.40. Only one of the outdoor collocated PM2.5 monitors (PEM) provided mass concentration data that were statistically different from that produced by a protoype PM2.5 FRM sampler. The PEM had a positive mass concentration bias ranging up to 18% relative to the FRM prototype. PMID- 11051540 TI - "Don't know much bile-ology". PMID- 11051541 TI - TBP-like factors come into focus. PMID- 11051542 TI - Noncoding RNA genes in dosage compensation and imprinting. PMID- 11051543 TI - Receptor tyrosine kinases: specific outcomes from general signals. PMID- 11051544 TI - Disruption of imprinted X inactivation by parent-of-origin effects at Tsix. AB - In marsupials and in extraembryonic tissues of placental mammals, X inactivation is imprinted to occur on the paternal chromosome. Here, we find that imprinting is controlled by the antisense Xist gene, Tsix. Tsix is maternally expressed and mice carrying a Tsix deletion show normal paternal but impaired maternal transmission. Maternal inheritance occurs infrequently, with surviving progeny showing intrauterine growth retardation and reduced fertility. Transmission ratio distortion results from disrupted imprinting and postimplantation loss of mutant embryos. In contrast to effects in embryonic stem cells, deleting Tsix causes ectopic X inactivation in early male embryos and inactivation of both X chromosomes in female embryos, indicating that X chromosome counting cannot override Tsix imprinting. These results highlight differences between imprinted and random X inactivation but show that Tsix regulates both. We propose that an imprinting center lies within Tsix. PMID- 11051545 TI - A mechanism for translationally coupled mRNA turnover: interaction between the poly(A) tail and a c-fos RNA coding determinant via a protein complex. AB - mRNA turnover mediated by the major protein-coding-region determinant of instability (mCRD) of the c-fos proto-oncogene transcript illustrates a functional interplay between mRNA turnover and translation. We show that the function of mCRD depends on its distance from the poly(A) tail. Five mCRD associated proteins were identified: Unr, a purine-rich RNA binding protein; PABP, a poly(A) binding protein; PAIP-1, a poly(A) binding protein interacting protein; hnRNP D, an AU-rich element binding protein; and NSAP1, an hnRNP R-like protein. These proteins form a multiprotein complex. Overexpression of these proteins stabilized mCRD-containing mRNA by impeding deadenylation. We propose that a bridging complex forms between the poly(A) tail and the mCRD and ribosome transit disrupts or reorganizes the complex, leading to rapid RNA deadenylation and decay. PMID- 11051546 TI - The osteoclast differentiation factor osteoprotegerin-ligand is essential for mammary gland development. AB - Osteoprotegerin-ligand (OPGL) is a key osteoclast differentiation/activation factor essential for bone remodeling. We report that mice lacking OPGL or its receptor RANK fail to form lobulo-alveolar mammary structures during pregnancy, resulting in death of newborns. Transplantation and OPGL-rescue experiments in opgl-/- and rank-/- pregnant females showed that OPGL acts directly on RANK expressing mammary epithelial cells. The effects of OPGL are autonomous to epithelial cells. The mammary gland defect in female opgl-/- mice is characterized by enhanced apoptosis and failures in proliferation and PKB activation in lobulo-alveolar buds that can be reversed by recombinant OPGL treatment. These data provide a novel paradigm in mammary gland development and an evolutionary rationale for hormonal regulation and gender bias of osteoporosis in females. PMID- 11051547 TI - D-cbl, a negative regulator of the Egfr pathway, is required for dorsoventral patterning in Drosophila oogenesis. AB - During Drosophila oogenesis, asymmetrically localized Gurken activates the EGF receptor (Egfr) and determines dorsal follicle cell fates. Using a mosaic follicle cell system we have identified a mutation in the D-cbl gene which causes hyperactivation of the Egfr pathway. Cbl proteins are known to downregulate activated receptors. We find that the abnormal Egfr activation is ligand dependent. Our results show that the precise regulation of Egfr activity necessary to establish different follicle cell fates requires two levels of control. The localized ligand Gurken activates Egfr to different levels in different follicle cells. In addition, Egfr activity has to be repressed through the activity of D-cbl to ensure the absence of signaling in the ventral most follicle cells. PMID- 11051548 TI - Ras pathway specificity is determined by the integration of multiple signal activated and tissue-restricted transcription factors. AB - Ras signaling elicits diverse outputs, yet how Ras specificity is generated remains incompletely understood. We demonstrate that Wingless (Wg) and Decapentaplegic (Dpp) confer competence for receptor tyrosine kinase-mediated induction of a subset of Drosophila muscle and cardiac progenitors by acting both upstream of and in parallel to Ras. In addition to regulating the expression of proximal Ras pathway components, Wg and Dpp coordinate the direct effects of three signal-activated (dTCF, Mad, and Pointed-functioning in the Wg, Dpp, and Ras/MAPK pathways, respectively) and two tissue-restricted (Twist and Tinman) transcription factors on a progenitor identity gene enhancer. The integration of Pointed with the combinatorial effects of dTCF, Mad, Twist, and Tinman determines inductive Ras signaling specificity in muscle and heart development. PMID- 11051549 TI - Combinatorial signaling in the specification of unique cell fates. AB - How multifunctional signals combine to specify unique cell fates during pattern formation is not well understood. Here, we demonstrate that together with the transcription factor Lozenge, the nuclear effectors of the EGFR and Notch signaling pathways directly regulate D-Pax2 transcription in cone cells of the Drosophila eye disc. Moreover, the specificity of D-Pax2 expression can be altered upon genetic manipulation of these inputs. Thus, a relatively small number of temporally and spatially controlled signals received by a set of pluripotent cells can create the unique combinations of activated transcription factors required to regulate target genes and ultimately specify distinct cell fates within this group. We expect that similar mechanisms may specify pattern formation in vertebrate developmental systems that involve intercellular communication. PMID- 11051550 TI - Overlapping activators and repressors delimit transcriptional response to receptor tyrosine kinase signals in the Drosophila eye. AB - Regulated transcription of the prospero gene in the Drosophila eye provides a model for how gene expression is specifically controlled by signals from receptor tyrosine kinases. We show that prospero is controlled by signals from the EGF receptor DER and the Sevenless receptor. A direct link is established between DER activation of a transcription enhancer in prospero and binding of two transcription factors that are targets of DER signaling. Binding of the cell specific Lozenge protein is also required for activation, and overlapping Lozenge protein distribution and DER signaling establishes expression in a subset of equivalent cells competent to respond to Sevenless. We show that Sevenless activates prospero independent of the enhancer and involves targeted degradation of Tramtrack, a transcription repressor. PMID- 11051551 TI - ICEBERG: a novel inhibitor of interleukin-1beta generation. AB - ProIL-1beta is a proinflammatory cytokine that is proteolytically processed to its active form by caspase-1. Upon receipt of a proinflammatory stimulus, an upstream adaptor, RIP2, binds and oligomerizes caspase-1 zymogen, promoting its autoactivation. ICEBERG is a novel protein that inhibits generation of IL-1beta by interacting with caspase-1 and preventing its association with RIP2. ICEBERG is induced by proinflammatory stimuli, suggesting that it may be part of a negative feedback loop. Consistent with this, enforced retroviral expression of ICEBERG inhibits lipopolysaccharide-induced IL-1beta generation. The structure of ICEBERG reveals it to be a member of the death-domain-fold superfamily. The distribution of surface charge is complementary to the homologous prodomain of caspase-1, suggesting that charge-charge interactions mediate binding of ICEBERG to the prodomain of caspase-1. PMID- 11051552 TI - A signal transduction system that responds to extracellular iron. AB - Iron is essential for all organisms but can be toxic in excess. Iron homeostasis is typically regulated by cytoplasmic iron binding proteins, but here we describe a signal transduction system (PmrA/PmrB) that responds to extracytoplasmic ferric iron. Iron promoted transcription of PmrA-activated genes and resistance to the antibiotic polymyxin in Salmonella. The PmrB protein bound iron via its periplasmic domain which harbors two copies of the sequence ExxE, a motif present in the Saccharomyces FTR1 iron transporter and in mammalian ferritin light chain. A pmrA mutant was hypersensitive to killing by iron but displayed wild-type resistance to a variety of oxidants, suggesting PmrA/PmrB controls a novel pathway mediating the avoidance of iron toxicity. PMID- 11051553 TI - Nucleophosmin/B23 is a target of CDK2/cyclin E in centrosome duplication. AB - In animal cells, duplication of centrosomes and DNA is coordinated. Since CDK2/cyclin E triggers initiation of both events, activation of CDK2/cyclin E is thought to link these two events. We identified nucleophosmin (NPM/B23) as a substrate of CDK2/cyclin E in centrosome duplication. NPM/B23 associates specifically with unduplicated centrosomes, and NPM/B23 dissociates from centrosomes by CDK2/cyclin E-mediated phosphorylation. An anti-NPM/B23 antibody, which blocks this phosphorylation, suppresses the initiation of centrosome duplication in vivo. Moreover, expression of a nonphosphorylatable mutant NPM/ B23 in cells effectively blocks centrosome duplication. Thus, NPM/B23 is a target of CDK2/cyclin E in the initiation of centrosome duplication. PMID- 11051554 TI - Oligomeric tubulin in large transporting complex is transported via kinesin in squid giant axons. AB - Slow axonal transport depends on an active mechanism that conveys cytosolic proteins. To investigate its molecular mechanism, we now constructed an in vitro experimental system for observation of tubulin transport, using squid giant axons. After injecting fluorescence-labeled tubulin into the axons, we monitored the movement of fluorescence by confocal laser scanning microscopy and fluorescence correlation spectroscopy. Here, from the pharmacological experiments and the functional blocking of kinesin motor protein by anti-kinesin antibody, we show that the directional movement of fluorescent profile was dependent on kinesin motor function. The fluorescent correlation function and estimated translational diffusion time revealed that tubulin molecule was transported in a unique form of large transporting complex distinct from those of stable polymers or other cytosolic protein. PMID- 11051555 TI - A viral movement protein prevents spread of the gene silencing signal in Nicotiana benthamiana. AB - In plants, viruses induce an RNA-mediated defense that is similar to posttranscriptional gene silencing (PTGS) of transgenes. Here we demonstrate with potato virus X (PVX) that PTGS operates as a systemic, sequence-specific defense system. However, in grafting experiments or with movement defective forms of PVX, we could not detect systemic silencing unless the 25 kDa viral movement protein (p25) was made nonfunctional. Investigation of p25 revealed two branches to the PTGS pathway that converge in the production of 25 nucleotide RNAs corresponding to the target RNA. One of these branches is unique to virus-induced PTGS and is not affected by p25. The second branch is common to both virus- and transgene induced PTGS, is blocked by p25, and is likely to generate the systemic silencing signal. PMID- 11051556 TI - Accessory protein facilitated CFTR-CFTR interaction, a molecular mechanism to potentiate the chloride channel activity. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) gene encodes a chloride channel protein that belongs to the superfamily of ATP binding cassette (ABC) transporters. Phosphorylation by protein kinase A in the presence of ATP activates the CFTR-mediated chloride conductance of the apical membranes. We have identified a novel hydrophilic CFTR binding protein, CAP70, which is also concentrated on the apical surfaces. CAP70 consists of four PDZ domains, three of which are capable of binding to the CFTR C terminus. Linking at least two CFTR molecules via cytoplasmic C-terminal binding by either multivalent CAP70 or a bivalent monoclonal antibody potentiates the CFTR chloride channel activity. Thus, the CFTR channel can be switched to a more active conducting state via a modification of intermolecular CFTR-CFTR contact that is enhanced by an accessory protein. PMID- 11051557 TI - Multicopy molecular dynamics simulations suggest how to reconcile crystallographic and product formation data for camphor enantiomers bound to cytochrome P-450cam. AB - Multiple ligand binding modes are possible in many enzyme active sites; their presence in cytochrome P450cam (P450cam) is evident from crystallographic studies of the binding of thiocamphor and phenylimidazoles. Here, we use multicopy molecular dynamics simulations to compare the binding modes of (1R)- and (1S) camphor in the active site of P450cam. Simulations with (1R)-camphor, the natural substrate, serve to calibrate our protocol: 19 out of 20 copies of (1R)-camphor converged to coordinates very close to those observed for (1R)-camphor in its crystallographic complex with P450cam during the simulations. Simulations with the (1S)-camphor enantiomer showed greater mobility of the substrate, consistent with spectroscopic data, and resulted in 3 major binding modes. One of these is similar to the major conformation (of the two conformations assigned) in a recently determined crystal structure, but this conformation is not correctly oriented for regiospecific hydroxylation at C-5. The simulations, however, provide evidence for reorientation of (1S)-camphor upon formation of the reactive Fe-O intermediate to an orientation suitable for hydroxylation. The simulations thus permit rationalisation of the apparent inconsistency between the crystal structure and the reaction products. PMID- 11051558 TI - Mapping the active site polarity in structures of endothelial nitric oxide synthase heme domain complexed with isothioureas. AB - Analyzing the active site topology and plasticity of nitric oxide synthase (NOS) and understanding enzyme-drug interactions are crucial for the development of potent, isoform-selective NOS inhibitors. A small hydrophobic pocket in the active site is identified in the bovine eNOS heme domain structures complexed with potent isothiourea inhibitors: seleno analogue of S-ethyl-isothiourea, S isopropyl-isothiourea, and 2-aminothiazoline, respectively. These structures reveal the importance of nonpolar van der Waals contacts in addition to the well known hydrogen bonding interactions between inhibitor and enzyme. The scaffold of a potent NOS inhibitor should be capable of donating hydrogen bonds to as well as making nonpolar contacts with amino acids in the NOS active site. PMID- 11051559 TI - Roles of the axial push effect in cytochrome P450cam studied with the site directed mutagenesis at the heme proximal site. AB - To examine the roles of the axial thiolate in cytochrome P450-catalyzed reactions, a mutant of cytochrome P450cam, L358P, was prepared to remove one of the conserved amide protons that are proposed to neutralize the negative charge of the thiolate sulfur. The increased push effect of the thiolate in L358P was evidenced by the reduced reduction potential of the heme. The 15N-NMR and resonance Raman spectra of the mutant in the ferric-CN and in the ferrous-CO forms, respectively, also supported the increased push effect. The maintenance of stereo- and regioselectivities for d-camphor hydroxylation by the mutant suggests the minimum structural change at the distal site. The heterolysis/homolysis ratios of cumene hydroperoxide were the same for wild-type and L358P. However, we observed the enhanced monooxygenations of the unnatural substrates using dioxygen and electrons supplied from the reconstituted system, which indicate the significant role of the push effect in dioxygen activation. We interpret that the enhanced push effect inhibits the protonation of the inner oxygen atom and/or promotes the protonation of the outer oxygen atom in the putative iron hydroperoxo intermediate (Fe3+ -O-OH) of P450cam. This work is the first experimental indication of the significance of the axial cysteine for the P450 reactivity. PMID- 11051560 TI - Stopped-flow spectrophotometric analysis of intermediates in the peroxo-dependent inactivation of cytochrome P450 by aldehydes. AB - The reaction of hydrogen peroxide and certain aromatic aldehydes with cytochrome P450BM3-F87G results in the covalent modification of the heme cofactor of this monooxygenase. Analysis of the resulting heme by electronic absorption spectrophotometry indicates that the reaction in the BM3 isoform is analogous to that in P450(2B4), which apparently occurs via a peroxyhemiacetal intermediate [Kuo et al., Biochemistry, 38 (1999) 10511]. It was observed that replacement of the Phe-87 in the P450BM3 by the smaller glycyl residue was essential for the modification to proceed, as the wild-type enzyme showed no spectral changes under identical conditions. The kinetics of this reaction were examined by stopped-flow spectrophotometry with 3-phenylpropionaldehyde and 3-phenylbutyraldehyde as reactants. In each case, the process of heme modification was biphasic, with initial bleaching of the Soret absorbance, followed by an increase in absorbance centered at 430 nm, consistent with meso-heme adduct formation. The intermediate formed during phase I also showed an increased absorbance between 700 and 900 nm, relative to the native heme and the final product. Phase I showed a linear dependence on peroxide concentration, whereas saturation kinetics were observed for phase II. All of these observations are consistent with a mechanism involving radical attack at the gamma-meso position of the heme cofactor, resulting in the intermediate formation of an isoporphyrin, the deprotonation of which produces the gamma-meso-alkyl heme derivative. PMID- 11051561 TI - Transsulfuration in Saccharomyces cerevisiae is not dependent on heme: purification and characterization of recombinant yeast cystathionine beta synthase. AB - Cystathionine beta-synthase [CBS; L-serine hydro-lyase (adding homocysteine), EC 4.2.1.22] catalyzes the first committed step of transsulfuration in both yeast and humans. It has been established previously that human CBS is a hemeprotein but although the heme group appears to be essential for CBS activity, the exact function of the heme group is unknown. CBS activity is absent in heme deficient strains of Saccharomyces cerevisiae grown without heme supplementation. CBS activity can be restored by supplementing these strains with heme, implying that there is a heme requirement for yeast CBS. We subcloned, overexpressed and purified yeast CBS. The yeast enzyme shows absolute pyridoxal 5'-phosphate (PLP) dependence for activity but we could find no evidence for the presence of a heme group. Given the degree of sequence and mechanistic similarity between yeast and human CBS, this result indicates that heme is unlikely to play a direct catalytic role in the human CBS reaction mechanism. Further characterization revealed that, in contrast to human CBS, S-adenosylmethionine (AdoMet) does not activate yeast CBS. Yeast CBS was found to be coordinately regulated with proliferation in S. cerevisiae. This finding is the most likely explanation of the observed apparent heme dependence of transsulfuration in vivo. PMID- 11051562 TI - Proximal ligand control of heme iron coordination structure and reactivity with hydrogen peroxide: investigations of the myoglobin cavity mutant H93G with unnatural oxygen donor proximal ligands. AB - The role of the proximal heme iron ligand in activation of hydrogen peroxide and control of spin state and coordination number in heme proteins is not yet well understood. Although there are several examples of amino acid sidechains with oxygen atoms which can act as potential heme iron ligands, the occurrence of protein-derived oxygen donor ligation in natural protein systems is quite rare. The sperm whale myoglobin cavity mutant H93G Mb (D. Barrick, Biochemistry 33 (1994) 6546) has its proximal histidine ligand replaced by glycine, a mutation which leaves an open cavity capable of accommodation of a variety of unnatural potential proximal ligands. This provides a convenient system for studying ligand protein interactions. Molecular modeling of the proximal cavity in the active site of H93G Mb indicates that the cavity is of sufficient size to accommodate benzoate and phenolate in conformations that allow their oxygen atoms to come within binding distance of the heme iron. In addition, benzoate may occupy the cavity in an orientation which allows one carboxylate oxygen atom to ligate to the heme iron while the other carboxylate oxygen is within hydrogen bonding distance of serine 92. The ferric phenolate and benzoate complexes have been prepared and characterized by UV-visible and MCD spectroscopies. The benzoate adduct shows characteristics of a six-coordinate high-spin complex. To our knowledge, this is the first known example of a six-coordinate high-spin heme complex with an anionic oxygen donor proximal ligand. The benzoate ligand is displaced at alkaline pH and upon reaction with hydrogen peroxide. The phenolate adduct of H93G Mb is a five-coordinate high-spin complex whose UV-visible and MCD spectra are distinct from those of the histidine 93 to tyrosine (H93Y Mb) mutant of sperm whale myoglobin. The phenolate adduct is stable at alkaline pH and exhibits a reduced reactivity with hydrogen peroxide relative to that of both native ferric myoglobin, and the exogenous ligand-free derivative of ferric H93G Mb. These observations indicate that the identity of the proximal oxygen donor ligand has an important influence on both the heme iron coordination number and the reactivity of the complex with hydrogen peroxide. PMID- 11051563 TI - Microsomal cytochrome P450 2C5: comparison to microbial P450s and unique features. AB - Although microsomal P450s represent the majority of P450s, only microbial P450s have been amenable to crystal structure solution. We have recently solved the first crystal structure of a microsomal P450, 2C5, a progesterone hydroxylase from rabbit. We discuss the features of the protein in common with existing structures of microbial P450s and limitations of homology modeling mammalian P450s based on the microbial structures. Unique features involving membrane, substrate and cytochrome P450 reductase interactions are also discussed. PMID- 11051564 TI - Crystal structures of cytochrome P450nor and its mutants (Ser286-->Val, Thr) in the ferric resting state at cryogenic temperature: a comparative analysis with monooxygenase cytochrome P450s. AB - Cytochrome P450nor (P450nor) is a heme enzyme isolated from the denitrifying fungus Fusarium oxysporum and catalyzes the NO reduction to N2O. Crystal structures of the wild type and two Ser286 mutants (Ser286-->Val, Ser286-->Thr) of P450nor have been determined for the ferric resting forms at a 1.7 A resolution at cryogenic temperature (100 K). We carried out three comparative analyses: (1) between the structures of P450nor at room temperature and cryogenic temperature, (2) between the structures of P450nor and four monooxygenase P450s, and (3) between the structures of the WT and the Ser286 mutant enzymes of P450nor. Comparison of the charge distribution on the protein surface suggests that proton and electron flow to the heme site is quite different in P450nor than in monooxygenase P450s. On the basis of the mutant structures, it was found that a special hydrogen-bonding network, Wat99-Ser286-Wat39-Asp393-solvent, acts as a proton delivery pathway in NO reduction by P450nor. In addition, the positively charged cluster located beneath the B'-helix is suggested as possible NADH binding site in P450nor, from which the direct two-electron transfer to the heme site allows to generate the characteristic intermediate in the NO reduction. These structural characteristics were not observed in structures of monooxygenase P450s, implying that these are factors determining the unique NO reduction activity of P450nor. PMID- 11051565 TI - The role of tetrahydrobiopterin in the activation of oxygen by nitric-oxide synthase. AB - We have studied the reaction of reduced nitric-oxide synthase (NOS) with molecular oxygen at -30 degrees C. In the first reaction cycle (from L-Arg to hydroxy-L-Arg), an oxygen adduct complex formed rapidly. Experiments in the absence of the reductase domain demonstrated that this complex was then further reduced by one electron stemming from the cofactor tetrahydrobiopterin (BH4). Spectral evidence suggested an iron(IV) porphyrin pi-cation radical as an intermediate. The nature of the oxidized BH4 was identified by EPR as a BH3* radical. Within the second cycle (from hydroxy-L-Arg to citrulline and NO), an iron(III)-NO complex could be identified clearly by its spectral characteristics. The strict requirement of BH4 for its formation suggests that BH4 plays a redox role, although transient, also in the second reaction cycle. PMID- 11051566 TI - Autocatalytic nitration of P450CAM by peroxynitrite. AB - Peroxynitrite (PN) gains high selectivity as a physiological oxidizing and nitrating agent through catalysis by metal ions. This was established for the heme-thiolate (P450) enzyme prostacyclin synthase which was tyrosine nitrated and inhibited at low PN levels [FEBS Lett. 382 (1996) 101]. Other P450 proteins reacted in a similar manner and a ferryl species (Compound II) has been identified as an intermediate during reactions with PN [Nitric Oxide 3 (1999) 142]. Here we investigated cytochrome P450CAM and found that it catalyzes the decomposition of PN as well as an increased nitration of phenol. The latter at the expense of phenol hydroxylation is characteristic for the proton-assisted PN action. PN also caused self-nitration of P450CAM at several tyrosine residues. Two of them, Y96 and Y305 were largely protected in the presence of the ligand metyrapone. Unlike other heme-thiolate proteins P450CAM did not form distinct spectral intermediates characteristic for Compound II. We conclude that P450CAM serves as a model for the nitration of prostacyclin synthase with respect to its autocatalytic tyrosine nitration and its prevention by blocking the active site. PMID- 11051567 TI - Enantiomeric discrimination of Ru-substrates by cytochrome P450cam. AB - Molecules with photosensitizers attached to substrates (Wilker et al., Angew. Chem. Int. Ed. 38 (1999) 90-92) or cofactors (Hamachi et al., J. Am. Chem. Soc. 121 (1999) 5500-5506) can rapidly deliver redox equivalents to buried active sites. The structure of cytochrome P450cam (P450) co-crystallized with a prototypal sensitizer-substrate, [Ru-C9-Ad]Cl2, has been determined (Dmochowski et al., Proc. Natl. Acad. Sci. USA 96 (1999) 12987-12990); and, in separate UV vis absorption and time-resolved luminescence experiments, the binding of the lambda and delta enantiomers of Ru-C9-Ad to P450 has been measured. The results, KD(delta/lambda) approximately 2, indicate that the bipyridyl ligands of the lambda isomer interact more favorably with hydrophobic residues at the entrance to the substrate channel. We conclude that enantiospecific interactions may be exploited in the design of enzyme-metallosubstrate conjugates. PMID- 11051568 TI - Increasing intervention implementation in general education following consultation: a comparison of two follow-up strategies. AB - This study examined two strategies for increasing the accuracy with which general education teachers implemented a peer tutoring intervention for reading comprehension. The intervention was implemented for 5 elementary school students who had been referred for consultation services. Initial implementation of the intervention by the teachers was variable, and the data exhibited a downward trend. When consultants held brief daily meetings with the teachers to discuss the intervention, implementation improved for 2 of 5 participants. Four of the teachers implemented the intervention at levels substantially above baseline during the performance feedback condition, whereas implementation for 1 teacher increased following discussion of an upcoming follow-up meeting with the principal. Student reading comprehension scores improved markedly during the peer tutoring intervention. Three students maintained these gains 4 weeks after the intervention ended. The implications of these findings for the maintenance of accurate treatment implementation in applied settings are discussed. PMID- 11051569 TI - A component analysis of "stereotypy as reinforcement" for alternative behavior. AB - Results from several studies have suggested that the opportunity to engage in stereotypic behavior may function as reinforcement for alternative, more socially desirable behaviors. However, the procedural components of this intervention include several distinct operations whose effects have not been analyzed separately. While measuring the occurrence of stereotypy and an alternative behavior (manipulation of leisure materials), we exposed 3 participants to three or four components of a "stereotypy as reinforcement" contingency: (a) continuous access to materials, (b) prompts to manipulate materials, (c) restricted access to stereotypy (i.e., response blocking), and (d) access to stereotypy contingent on manipulating the materials. Continuous access to materials and prompting (a and b) produced negligible results. Restriction of stereotypy (c) produced a large increase in the alternative behavior of 2 participants, suggesting that response restriction per se may occasion alternative behavior. However, contingent access to stereotypy (d) was necessary to increase the 3rd participant's object manipulation; this finding provided some support for the use of stereotypy as reinforcement for alternative behavior. Finally, when transfer of the effects of intervention was assessed during periods in which active intervention components were withdrawn, the alternative behavior was maintained for 1 participant. PMID- 11051570 TI - Differential responding in the presence and absence of discriminative stimuli during multielement functional analyses. AB - We evaluated the extent to which discriminative stimuli (S(D)s) facilitate differential responding during multielement functional analyses. Eight individuals, all diagnosed with mental retardation and referred for assessment and treatment of self-injurious behavior (SIB) or aggression, participated. Functional analyses consisted of four or five assessment conditions alternated in multielement designs. Each condition was initially correlated with a specific therapist and a specific room color (S(D)s), and sessions continued until higher rates of target behaviors were consistently observed under a specific test condition. In a subsequent analysis, the programmed S(D)s were removed (i.e., all conditions were now conducted by the same therapist in the same room), and sessions continued until differential responding was observed or until twice as many sessions were conducted with the S(D)s absent (as opposed to present), whichever came first. Results indicated that the inclusion of programmed S(D)s facilitated discrimination among functional analysis conditions for half of the participants. These results suggest that the inclusion of salient cues may increase either the efficiency of functional analyses or the likelihood of obtaining clear assessment outcomes. PMID- 11051571 TI - Identity matching of consonant-vowel-consonant words by prereaders. AB - Using an identity matching-to-sample procedure, normally developing prereaders who matched individual letters with high accuracy (e.g., m and s) did not show high accuracy in matching three-letter printed words that differed only in the first letter (e.g., mad and sad). Teachers and researchers should not assume that children who can discriminate individual letters can also discriminate minimally different words that contain those letters. PMID- 11051572 TI - Extinction effects during the assessment of multiple problem behaviors. AB - Extinction effects were evaluated in a multiple baseline across behaviors design with 2 boys after just one of several target problem behaviors was observed during a functional analysis. Other target behaviors emerged as extinction was introduced sequentially across all problem behaviors. Results demonstrated an efficient strategy for simultaneously assessing multiple problem behaviors maintained by the same consequence. PMID- 11051573 TI - The blocking effect of pictorial prompts on sight-word reading. AB - This study replicates and extends previous work showing that pictorial prompts can interfere with the learning of sight words by students with moderate mental retardation. Effects of training with 6 students were assessed during five conditions using an alternating treatments design. In four conditions, words were presented either alone or with a corresponding picture. In a fifth condition, pictures were used to provide feedback. The results showed that acquisition was achieved fastest during the word-alone conditions with 5 students. PMID- 11051574 TI - Comparison of single and multiple functional communication training responses for the treatment of problem behavior. AB - Two functional communication training (FCT) conditions without extinction were compared to treat the problem behavior of a child with developmental disabilities. The individual was taught to emit a single FCT response to obtain one of six items delivered in a randomized order or multiple FCT responses that specified the exact item. Results showed that only the FCT-multiple condition reduced problem behavior and maintained alternative mands. PMID- 11051575 TI - Independent use of activity materials by the elderly in a residential setting. AB - A lottery was implemented to encourage the elderly clients of a residential home to use activity materials any time they wished, independently of staff intervention and the institutional routine of the home. During baseline, there were minimal levels of independent use of activity materials by residents. Various conditions were implemented but only the introduction of a l20 lottery prize brought about a significant increase in the frequency of independent use of activity materials. A follow-up suggested that the reinforcing properties of the activity materials themselves eventually maintained the target behavior. PMID- 11051576 TI - A comparison of verbal and tangible stimulus preference assessments. AB - Tangible preference assessments were compared with verbal preference assessments for 6 individuals with mental retardation, behavior disorders, or both. In the tangible assessment, items were placed in front of the participant. In the verbal assessment, participants were asked, "Do you want X or Y?" and the items were not present. The two assessments yielded similar high-preference items for 4 of the 6 participants. The verbal assessment was typically completed in less time than the tangible assessment. PMID- 11051577 TI - Further evaluation of the accuracy of reinforcer surveys: a systematic replication. AB - The present report evaluates the accuracy of a reinforcer survey by comparing the survey results to the results of subsequent reinforcer assessments for 20 children using a concurrent-operants arrangement to assess relative reinforcer preference. Total accuracy for the survey was determined to be approximately 57%. The results provide a systematic replication of Northup et al. (1996) with a much larger sample of children. A need for the development of more accurate and comprehensive reinforcer assessment methods for verbal children is discussed. PMID- 11051578 TI - International publication trends of JABA authorship. AB - The present study addressed international publication trends in JABA authorship between 1970 and 1999. First, we analyzed authorship patterns to identify trends in the appearance of new first authors, unfamiliar authors, and frequent contributors. Second, articles were assigned to either a North American or an international category. The data show a decline in the number of articles by new authors and an increase in the publications of frequent contributors from North America. Trends are shown in comparison to those from the American Journal on Mental Retardation. PMID- 11051579 TI - Noncontingent peer attention as treatment for disruptive classroom behavior. AB - A functional analysis isolated peer attention as the primary maintaining variable for disruptive behavior displayed by a student with attention deficit hyperactivity disorder. Using a brief reversal design, noncontingent reinforcement was then shown to reduce disruptive behavior relative to the peer attention condition. Implications for assessing behavior disorders in mainstream school settings are discussed. PMID- 11051580 TI - Effects of reinforcer choice measured in single-operant and concurrent-schedule procedures. AB - The effects of choice and no choice of reinforcer on the response rates of 3 children with autism were compared across single-operant and concurrent-schedule procedures. No consistent differences in responding between choice and no-choice components emerged during single-operant phases. During the concurrent-schedule phases, however, all participants had substantially higher rates of responding to the button that led to a choice among reinforcers than to the button that did not lead to choice. PMID- 11051581 TI - Evaluation of a brief multiple-stimulus preference assessment in a naturalistic context. AB - We evaluated a brief multiple-stimulus preference assessment within the context of an early intervention program for 3 children who had been diagnosed with autism. Subsequent curriculum-based reinforcer evaluations confirmed the predictions of the preference assessments. In addition, eight additional preference assessments that were conducted over a period of 1 month indicated generally stable preferences for 2 of the 3 participants. PMID- 11051583 TI - The problem of parental nonadherence in clinical behavior analysis: effective treatment is not enough. AB - Applied behavior analysts have developed many effective interventions for common childhood problems and have repeatedly demonstrated that childhood behavior responds to properly managed contingencies. The success of these interventions is dependent upon their basic effectiveness, as demonstrated in the literature, their precise delivery by the clinician to the parent, and adherence to or consistent implementation of the intervention. Unfortunately, arranging the consistent implementation of effective parenting strategies is a significant challenge for behavior analysts who work in homes, schools, and outpatient or primary care clinics. Much has been done to address issues of adherence or implementation in the clinic, but relatively little has been done to increase our understanding of the contingencies that affect parental adherence beyond the supervised clinic environment. An analysis of the contingencies that strengthen or weaken adherence might suggest strategies to improve implementation outside the clinic setting. What follows is an analysis of the variables associated with adherence by parents to recommendations designed to solve common childhood problems. PMID- 11051582 TI - Designing interventions that include delayed reinforcement: implications of recent laboratory research. AB - The search for robust and durable interventions in everyday situations typically involves the use of delayed reinforcers, sometimes delivered well after a target behavior occurs. Integrating the findings from laboratory research on delayed reinforcement can contribute to the design and analysis of those applied interventions. As illustrations, we examine articles from the Journal of the Experimental Analysis of Behavior that analyzed delayed reinforcement with respect to response allocation (A. M. Williams & Lattal, 1999), stimulus chaining (B. A. Williams, 1999), and self-control (Jackson & Hackenberg, 1996). These studies help to clarify the conditions under which delayed reinforcement (a) exercises control of behavior, (b) entails conditioned reinforcement, and (c) displaces the effects of immediate reinforcement. The research has applied implications, including the development of positive social behavior and teaching people to make adaptive choices. DESCRIPTORS: delayed reinforcement, response allocation, stimulus chains, self-control, integration of basic and applied research PMID- 11051584 TI - Seminal plasma trace metal levels in industrial workers. AB - This study compares the seminal plasma trace metal levels of hospital workers with groups of industrial workers in a petroleum refinery, smelter, and chemical plant. The metals measured were the essential metals (copper, zinc, nickel, cobalt, and manganese) and the toxic metals (lead, cadmium, and aluminum). The group mean +/- SE metal level for each group (50 subjects per group) was calculated, and the statistical significance of the group mean differences of the industrial groups with the hospital group (control) was determined by the Student's t-test. The differences observed in the smelter group were increased copper and zinc (p < or = 0.001) and decreased nickel, cobalt, and manganese (p < or = 0.001, < or = 0.01). The refinery group differences were increased copper, zinc, and nickel (p < or = 0.001) but decreased cobalt and manganese (p < or = 0.001). The chemical group differences were increased zinc (p < or = 0.001) and decreased cobalt (p < or = 0.001). The seminal plasma levels of the toxic metals lead and aluminum were increased in each of the industrial groups (p < or = 0.001). Concurrent differences were (1) decreased accumulation of nickel, cobalt, and manganese in the smelter group, (2) decreased cobalt and manganese in the refinery group, and (3) only decreased cobalt in the chemical group. PMID- 11051585 TI - Blood cell lead, calcium, and magnesium levels associated with pregnancy-induced hypertension and preeclampsia. AB - This study compares the red blood cell (Rbc) levels of lead (Pb), calcium (Ca), and magnesium (Mg) in relation to blood pressure in 39 pregnant women in the third trimester of pregnancy. The study population included 20 women with normal pregnancies, 15 with mild hypertension, and 4 with severe hypertension and preeclampsia. The mean +/- SD for each group was calculated and the difference between the means of the normotensive and the other groups were compared by analysis of variance. Significant differences from normal to the preeclamptic pregnancies were in (1) elevated Rbc Pb (p < or = 0.001), (2) lower Rbc Ca (p < or = 0.001), and (3) lower Rbc Mg/Pb ratio (p < or = 0.0001). Pearson's rank correlation between blood pressure showed a direct relation to the Rbc Pb level (p < or = 0.01) and an inverse relation to the Rbc Ca and Mg/Pb ratio (p < or = 0.004, < or = 0.007). Apparently, prenatal blood pressure is directly proportional to Rbc Pb content and related or modified by Rbc Ca and Mg. PMID- 11051586 TI - Age-related changes of elements in human ureter. AB - To elucidate changes of the ureter with aging, the authors investigated age related changes of element contents in human ureters. The subjects consisted of seven men and seven women, ranging in age from 61 to 97 yr. The contents of calcium, sulfur, and iron in the ureters increased progressively with aging, whereas the contents of phosphorus and magnesium did not increase with aging. Significant relationships were found both between calcium and sulfur contents and between calcium and iron contents in the ureters, but not between calcium and either phosphorus or magnesium contents. It was noteworthy that a significant relationship was also found between sulfur and iron contents in the ureters. It remains uncertain whether calcium forms a compound with sulfur or iron in aged human ureters or not. PMID- 11051587 TI - Comparative efficacy of several potential treatments for copper mobilization in copper-overloaded rats. AB - D-Penicillamine (DPA) is effective in the treatment of Wilson's disease, whereas zinc salts are also used as a therapy for this disorder of copper transport. Recently, it has been shown that the copper chelators 1,4,7,11-tetraazaundecane tetrahydrochloride (TAUD) and tetraethylenepentamine pentahydrochloride (TETREN) could be useful for copper mobilization in rats. Because these agents could be potential clinical alternatives to DPA for patients with Wilson's disease who are intolerant to this drug, we examined whether oral administration of TAUD and TETREN could be effective in mobilizing copper in experimental copper-overloaded rats. The efficacy of a combined administration of zinc and DPA, TAUD, or TETREN was also assessed. Rats were copper loaded with 0.125% copper acetate in water for 12 wk. After this period, DPA, TAUD, and TETREN were administered by gavage at 0.67 mmol/kg/d for 5 d, and zinc was given at 2.5 mg Zn/kg/d. Twelve weeks of copper loading resulted in a 32-fold increase in total hepatic copper. TETREN was the most effective chelator in increasing the urinary excretion of copper. However, it did not reduce significantly the hepatic copper levels. In turn, combined administration of zinc and chelating agents significantly reduced the amount of copper found in the feces. Although TAUD and TETREN showed a similar or higher efficacy to DPA in mobilizing copper, concurrent treatment of chelating agents and zinc salts should be discarded according to the current results. PMID- 11051588 TI - Nickel absorption and distribution from rat small intestine in situ. AB - The aim of this work was to study the absorption of nickel chloride in rats by means of the intestinal perfusion in situ technique at nickel concentrations of 1, 5, 10, 25, and 100 mg/L. Active transport and facilitated diffusion seem to play an important role in the intestinal absorption of nickel at concentrations < or = 10 mg/L. At higher concentrations, the absorption rate would be limited by saturation of the carriers. The distribution of the absorbed nickel was studied by intestinal perfusion of a 10-mg Ni/L solution for 30 or 60 min. Both in concentration and amount, the jejunum showed the higher values of absorbed nickel, followed by the kidneys and liver. When all of the collected organs (brain, heart, liver, lungs, spleen, kidneys, and testicles) and blood, but not the small intestine, are analyzed following a 60-min perfusion, it was found that 1% of the initial concentration had passed through the intestinal barrier. PMID- 11051589 TI - Cadmium-induced changes in parietal cell structure and functions of rats. AB - The aim of this study was to determine the cadmium (Cd)-induced functional and structural changes in gastric parietal cells of male rats exposed to high Cd for 30 d. In the present study, control animals were fed with normal food and tap water; the remaining animals received Cd (15 ppm CdCl2) in drinking water for the same period. Receiving Cd for 30 d increased the mean blood Cd level, the mean tissue Cd content, and the mean blood pressure (p < 0.01, p < 0.001, p < 0.01, respectively). The basal acid output fell; however, the increases in stimulated acid output were not statistically significant. Light and electron microscopic examination revealed respectively that (1) Cd decreases the mean parietal cell number per unit from the control value of 23.46 +/- 3.84 to 19.46 +/- 2.12 (p < 0.05) and it affected preferentially the cells located at the distal half of the zymogenic unit and (2) in parietal cells, the Cd-induced alterations were characterized with swollen canalicular profiles, broken-down tubulovesicles, or degenerated mitochondria. We concluded that Cd augments the elimination rate of parietal cells by increasing the alteration rate and reduced basal acid output can be explained easily with the loss of parietal cell population. PMID- 11051590 TI - Protective effects of zinc in chlorpyrifos induced hepatotoxicity: a biochemical and trace elemental study. AB - The toxicological literature is replete with studies attempting to explain the mechanism of action of organophosphorus (OP) insecticides to their anticholinesterase activities, but not much is known about the metabolism and detoxification of these compounds. The goal of this study was to ascertain the toxic effects of chlorpyrifos, one of the most widely used OPs, on the liver of male rats and also to evaluate the protective potential of zinc in mediating its toxic effects. It was observed that chlorpyrifos (13.5 mg/kg body weight) treatment resulted in significant inhibition (p < 0.001) of serum and hepatic acetylcholinesterase (AChE) activities after 8 wk. However, zinc-treated (227 mg/L drinking water) animals resulted in significant normalization of the inhibited AChE activities. Similarly, a significant increase in the levels of various serum and liver marker enzymes (viz. alkaline phosphatase, aspartate aminotransferase [AST], and alanine aminotransferase [ALT]) was observed following treatment with chlorpyrifos. However, coadministration of zinc to these animals restored these enzymes to within normal limits, even though some increase in the activity of serum ALT and hepatic alkaline phosphatase still persisted at the end of the study. Chlorpyrifos treatment diminished serum and hepatic zinc levels significantly (p < 0.01 to p < 0.001) compared to normal control animals. Serum iron concentrations also plummeted significantly following chlorpyrifos treatment. On the contrary, serum copper levels were significantly increased (p < 0.01) in chlorpyrifos-treated animals, but they were normalized following zinc supplementation to the rats in this group. Interestingly, chlorpyrifos treatment resulted in elevated hepatic levels of copper, iron, and selenium, but zinc treatment could only partially restore the raised elemental concentrations. These data clearly demonstrate the potential role of zinc in mediating the toxic effects of chlorpyrifos, presumably because of their antioxidant properties and also their possible interaction with other trace elements in maintaining the cellular harmony. PMID- 11051591 TI - Protective effects of zinc on cadmium toxicity in rodents. AB - A study of acute and subacute toxicity of cadmium ions [Cd(II)] was carried out on male Swiss mice and Sprague-Dawley rats with and without previous administration of zinc chloride. The LD50 of Cd(II) as cadmium sulfate (ip) was lower in animals previously given 10 mg/kg of zinc(II) chloride (sc). Factors such as animal weight variations, biochemical parameters, and accumulation patterns of Cd(II) and Zn(II) were taken into consideration when the subacute toxicity was evaluated. Alteration of the activities of glutamic pyruvic transaminase (GPT) and of glutamic oxaloacetic transaminase (GOT) was observed in short-term-exposure (<6 h) cases. These alterations reverted to normal after 1 wk. The activity of alkaline phosphatase (ALP) and the concentrations of cholesterol and triglycerides in serum are also changed, especially so in the groups given CdSO4 alone. In the experimental groups treated with ZnCl2 prior to administration of cadmium, proteinuria was detected 5 wk after the treatment. Also at 5 wk, both Zn-treated and nontreated groups showed an abnormally low liver mass with respect to total body mass. Both Cd and Zn are retained preferentially in the liver but show also in the kidneys. If CdSO4 and ZnCl2 are given simultaneously, especially after 1 wk of treatment, Cd is accumulated in greater amounts in these organs when compared to the groups given only cadmium sulfate. PMID- 11051592 TI - In vitro dialyzability of zinc from different salts used in the supplementation of infant formulas. AB - Seven zinc salts--acetate, chloride, lactate, sulfate, citrate, gluconate, and oxide--were added to milk--and soy-based infant formulas to estimate possible differences in zinc availability depending on the type of salt used. For this purpose, an in vitro method that estimates the dialyzability of the element (i.e., the fraction available for absorption) was applied. Zinc dialyzability is always higher in milk-based products than in soy products, even when the total zinc contents are higher in the latter. The salts can be classified according to the zinc dialyzability in the two types of formulas as follows: oxide > gluconate = chloride = lactate > citrate = acetate > sulfate. Therefore, according to the dialysis percentage, oxide and gluconate are the compounds of choice for zinc supplementation of infant formulas. PMID- 11051593 TI - Zinc and nitric oxide synthase inhibitor L-NAME attenuate NPY-induced feeding in mice. AB - The influences of zinc (Zn) and the nitric oxide synthase (NOS) inhibitor L-NAME on peripheral neuropeptide Y (NPY)-induced feeding in mice were investigated. Male mice received NPY (200 ng/d/mouse subcutaneously) and were separated into four groups based on cotreatments (with or without Zn [0.1 mg/mL]) and with or without L-NAME [0.2 mg/mL]) administered in drinking water for 10 d. A control group that received saline injection was also studied. The results showed that NPY, with or without any studied chemicals, did not affect body weight gain or body fat content. However, the mice that were administered NPY alone had increased energy intakes, higher serum triglyceride and free fatty acid, and lower serum glucose than saline-injected controls. NPY-treated mice that were given Zn and L-NAME cotreatments had compatible results of determined variables in comparison with control mice. This study showed that Zn and L-NAME attenuated NPY-mediated feeding and selected serum variables in mice. However, the mechanisms of the interactions among NPY, Zn and NOS, and their effects on appetite regulation, remain to be elucidated. PMID- 11051594 TI - Dietary molar ratios of phytate:zinc and millimolar ratios of phytate x calcium:zinc in South Koreans. AB - The zinc nutritional status in south Koreans was established by evaluation of zinc, calcium and phytate intakes, the molar ratio of phytate:zinc, and the millimolar ratio of phytate x calcium:zinc. The intakes of iron and magnesium were also estimated. Sampling was designed so that it was representative of the national population. Two-day food records were used for the calculation of nutrient intakes, using food consumption data from the 1995 National Nutrition Survey ('95NNS) for South Korea. Daily intakes of zinc and calcium were estimated to be 10.1 mg/d and 426.5 mg/d, respectively, and those of iron and magnesium were 15.2 mg/d and 268.0 mg/d, respectively. The estimated daily phytate intake was 1676.6 mg/d. The ratio of phytate:zinc was 15.9 mol/d and that of phytate x calcium:zinc was 168.9 mmol/d. The ratio in millimoles per 4.2 MJ (1000 kcal) of phytate x calcium:zinc was 91.8. The major food groups for zinc intake were meat, poultry and their products (43%), and cereals and grain products (18%). Sixty-two percent of zinc was from animal food sources. Cereal and grain products supplied most of the phytate intake (46%) followed by seasonings, fruits, and legumes and their products. The major food source of phytate was rice (39%). The results of the study raise concern about the suboptimal zinc status in relation to the Western diet. PMID- 11051595 TI - Zinc concentration of selected ocular tissues in zinc-deficient rats. AB - A study was performed to determine the effect of zinc deficiency on the zinc concentration of the retina, lens, and the retinal pigment epithelium and choroid. Weanling, male Sprague-Dawley rats were fed ad libitum modified AIN-93 diets containing 3 mg zinc/kg diet (-Zn; n = 10) for 6 wk. Control animals were pair-fed (+ZnPF; n = 10) or fed ad libitum (+ZnAL; n = 10) diets containing 100 mg zinc/kg diet. At 6 wk, plasma and tibia zinc were measured by flame atomic absorption spectrophotometry to confirm zinc deficiency. The zinc concentration of ocular tissues was measured by inductively coupled plasma-mass spectrometry. Mean (+/- SEM) lens zinc concentration was significantly depressed in the zinc deficient group as compared to that of pair-fed or ad libitum-fed controls, suggesting that the role of zinc in cataract formation should be investigated. The zinc concentration of total neural retina was preserved in zinc deficiency. Previously reported deterioration of retinal function in zinc deficiency may be the result of a decline in the zinc concentration of a specific cell layer of the retina that cannot be detected on gross analysis of the entire retina. PMID- 11051596 TI - Preliminary study of combination therapy with interferon-alpha and zinc in chronic hepatitis C patients with genotype 1b. AB - We have evaluated the efficacy of interferon-alpha (IFN-alpha) plus zinc therapy in hepatitis C patients with genotype 1b, poor responders for IFN alone. Ten patients were injected with 10 MU of IFN-alpha every day for 4 wk, followed by three times a week for 20 wk (control group). Nine patients took 300 mg of zinc sulfate a day orally during IFN-alpha therapy (zinc sulfate group), and 15 patients took IFN-alpha and 150 mg of polaprezinc (polaprezinc group). On the d 8 of IFN therapy, circadian zinc levels in serum elevated significantly in the polaprezinc group compared to the zinc sulfate group or control group. Serum ALT levels normalized in 73.3% of the polaprezinc group, 55.6% of the zinc sulfate group, and 40.0% of the control group at 6 mo after the end of IFN therapy. Sustained eradication for the hepatitis C virus RNA judged at the end of the 6-mo follow-up period was higher in the polaprezinc group than in the zinc sulfate group (53.3% vs 11.1%, p < 0.05) or the control group (20.0%). No clinical side effects of zinc were observed at the dose used. The data suggest that polaprezinc is expected to increase the therapeutic response of IFN-alpha for chronic hepatitis C with genotype 1b. PMID- 11051597 TI - Interrelationship of indices of body composition and zinc status in 11-yr-old New Zealand children. AB - Serum zinc and hair zinc concentrations of some New Zealand children aged 11 yr, were examined in relation to selected anthropometric indices. Serum zinc concentrations (n = 453) in boys and girls were similar and were unrelated to anthropometric indices and hair zinc concentrations. Mean hair zinc concentration (n = 620) of the girls was higher than that for the boys (2.95+/-0.49 vs 2.46+/ 0.47 micromol/g; p < 0.001). Correlation analysis demonstrated that, for the boys, all the studied anthropometric indices, with the exception of height, were significantly related to hair zinc concentration and that the confounding effects of mid-parent height and the timing of the adolescent growth spurt was small. Results for the girls were similar but less significant. Dichotomizing the hair zinc results divided both the boys and girls into two groups: those with hair zinc < 2.44 micromol/g were heavier (girls, 39.0 vs 35.2 kg; boys, 36.6 vs 34.7 kg) and fatter (mid-upper-arm fat area: girls, 15.2 vs 12.0 cm2; boys, 11.1 vs 9.5 cm2) compared to their counterparts with hair zinc > 2.44 micromol/g. The results demonstrate that in these healthy New Zealand children, those with lower hair zinc concentrations are fatter and heavier than their high-hair-zinc counterparts. PMID- 11051598 TI - Serum zinc levels and Alzheimer's disease. AB - Concentrations of zinc in postmortem serum and four brain regions were measured by flame atomic absorption spectrometry and instrumental neutron activation analysis, respectively, in nine Alzheimer's disease (AD) and eight control subjects. A statistically significant elevation of zinc serum was observed in AD subjects (136.4+/-66.8 microg/dL) compared with age-matched control subjects (71.1+/-35.0 microg/dL). No significant differences were observed between AD and control zinc concentrations in the amygdala, hippocampus, cerebellum, and superior and middle temporal gryi. PMID- 11051599 TI - Activation of glycolysis by zinc is diminished in hepatocytes from metallothionein-null mice. AB - The influence of hepatic metallothionein (MT) and zinc (Zn) on glycolysis was investigated in primary cultures of mouse hepatocytes prepared from MT-normal (+/+) and MT-null (-/-) mice. In MT +/+ mice, a close relationship was observed between the Zn concentration in the incubation medium (10-150 microM), increased MT levels in the cells, and increased glycolysis (accumulation of lactate + pyruvate) over 24 h, with significant effects seen at physiological levels of Zn (10-25 microM). Hepatocytes from MT -/- mice had significantly lower basal rates of glycolysis and demonstrated increased glycolysis only at Zn concentrations of 50 microM or greater. The lactate:pyruvate ratio was higher in the MT +/+ hepatocytes. The oxidation of endogenous fatty acid (accumulation of the ketone bodies, 3-hydroxybutyrate and acetoacetate) was initially greater in the MT +/+ hepatocytes, although only MT -/- hepatocytes showed increased ketone body production in response to Zn. The 3-hydroxybutyrate:acetoacetate ratio was higher in the MT +/+ hepatocytes and increased with increasing Zn concentrations. Intracellular Zn accumulation was 60% greater in the MT +/+ hepatocytes, with approximately 80% of the extra Zn associated with MT. The results implicate MT associated Zn rather than increased intracellular Zn per se in the regulation of hepatic carbohydrate metabolism. PMID- 11051600 TI - Leptin concentration and the Zn/Cu ratio in plasma in women with thyroid disorder. AB - We investigated the possible correlation between the leptin concentration and the Zn/Cu ratio in the plasma of women with thyroid disorder. Forty women with hypothyroidism (n = 20) or hypothyroidism (n = 20) and 20 euthyroid controls were recruited. The results showed that the women with thyroid disorder (hypothyroidism or hyperthyroidism) had higher plasma leptin concentrations than the normal controls (p < 0.05). Moreover, the plasma leptin concentration had no correlation with plasma thyroid hormone levels in the separate groups, nor among all the participants considered together. A strong correlation (p < 0.005) between leptin and adiposity was only observed in euthyroid women. Plasma values of Zn and Cu and the Zn/Cu ratio were not markedly different among women with altered thyroid status. However, a weak correlation (r = 0.28, p = 0.032) between leptin and the Zn/Cu ratio was found from the pooled data of all participants and retained after adjustment for adiposity. We suggest that there may exist an interaction between the plasma leptin level and thyroid hormone-induced abnormality for selected minerals. PMID- 11051601 TI - Leukocyte selenium, zinc, and copper concentrations in preeclamptic and normotensive pregnant women. AB - Preeclampsia is an important cause of maternal and perinatal mortality worldwide. The etiology of this relatively common medical complication of pregnancy, however, remains unknown. We studied the relationship between maternal leukocyte selenium, zinc, and copper concentrations and the risk of preeclampsia in a large hospital-based case-control study. One hundred seventy-one women with proteinuric pregnancy-induced hypertension (with or without seizures) comprised the case group. Controls were 184 normotensive pregnant women. Leukocytes were separated from blood samples collected during the patients' postpartum labor and delivery admission. Leukocyte concentrations for the three cations were measured by inductively coupled plasma-mass spectrometry (ICP-MS). Concentrations for each cation were reported as micrograms per gram of total protein. Women with preeclampsia had significantly higher median leukocyte selenium concentrations than normotensive controls (3.23 vs 2.80 microg/g total protein, p < 0.0001). Median leukocyte zinc concentrations were 31% higher in preeclamptics as compared with controls (179.15 vs 136.44 microg/g total protein, p < 0.0001). Although median leukocyte copper concentrations were slightly higher for cases than controls, this difference did not reach statistical significance (17.72 vs 17.00 microg/g total protein, p = 0.468). There was evidence of a linear increase in risk of preeclampsia with increasing concentrations of selenium and zinc. The relative risk for preeclampsia was 3.38 (adjusted odds ratio [OR] = 3.38, 95% confidence interval [CI] = 1.53-7.54) among women in the highest quartile of the control selenium distribution compared with women in the lowest quartile. The corresponding relative risk and 95% CI for preeclampsia was 5.30 (2.45-11.44) for women in the highest quartile of the control zinc distribution compared with women in the lowest quartile. There was no clear pattern of a linear trend in risk with increasing concentration of leukocyte copper concentrations (adjusted for linear trend in risk = 0.299). Our results are consistent with some previous reports. Prospective studies are needed to determine whether observed alterations in selenium and zinc concentrations precede preeclampsia or whether the differences may be attributed to preeclampsia-related alterations in maternal and fetal-placental trace metal metabolism. PMID- 11051602 TI - Selenium deficiency-induced alterations in the vascular system of the rat. AB - To enunciate the mechanisms whereby Se protects against cardiovascular diseases, weanling male Wistar rats were fed deficient (0.022 mg/kg diet) and adequate (0.159 mg/kg diet) Se diets for 14 and/or 39 wk. As the Se content and glutathione peroxidase activity were decreased and the lipid peroxide level was increased, the plasma 6-keto-PGF1alpha concentration of the Se-deficient group was markedly decreased in blood and tissues of the Se-deficient rats, as compared with the Se-adequate animals. Furthermore, the Se-deficient group had significantly lower plasma nitric oxide content and vascular nitric oxide synthase activity, higher erythrocyte sedimentation equation K value and aggregation index, and lower erythrocyte deformability than the Se-adequate group. Experimental Se deficiency also resulted in significant increases in serum total cholesterol and low-density lipoprotein cholesterol levels and a significant decrease in serum high-density lipoprotein cholesterol level. These results give some experimental supports to the hypothesis that low Se status and lipid peroxidation are involved in the etiology of cardiovascular diseases. PMID- 11051603 TI - Disulfonic stilbene prevents selenite-induced cataract in rat pup lens. AB - It is known that the subcutaneous injection of a single dose of sodium selenite into suckling rats results in the development of large nuclear opacities. The intracellular transport of selenite in various cells, except lens cells, occurs via the Cl/HCO3 exchanger. The aim of the present study is to investigate the possible role of the anion-exchange inhibitor, disulfonic stilbene (SITS), in the selenite-induced catarogenesis in the rat pups. Wistar albino rats (8-10 d old) were separated into three groups: one control and two experimental. The first experimental group was injected subcutaneously with a single dose of 30 nmol sodium selenite/g body weight. The second experimental group was injected with a single dose of 10 nmol SITS/g body weight 15 min before the same dose selenite injection. The control group did not have any injections. The stage of cataract development was examined on d 7 postinjection with slit-lamp photographs. In SITS pretreated group, all eyes remained transparent (considered as stage 0), whereas in the selenite-injected group, the animals did have different stage of nuclear cataract; 8 animals have stage 5, 10 animals have stage 4, and 4 animals have stage 3. A pretreatment of SITS completely prevented cataract formation of the selenite-induced cataract model in rat pups. PMID- 11051604 TI - Preliminary study on the determination of selenium compounds in some selenium accumulating mushrooms. AB - Using various chromatographic techniques (size exclusion, anion exchange, and cation exchange) combined with several detectors (neutron activation analysis and atomic fluorescence spectrometry), an attempt was made to characterize selenium compounds in some edible, selenium-accumulating mushrooms (Albatrellus pes-caprae and Boletus edulis). The mushrooms contained mostly low-molecular-weight (6 kDa) selenium compounds. After proteolysis, only a small fraction of the extractable selenium could be identified as selenite (3.0-9.2%, Albatrellus pes-caprae), selenocystine (minor, Albatrellus pes-caprae; 7.5%, Boletus edulis), or selenomethionine (1.0%, Boletus edulis), leaving the form of the bulk still to be elucidated. PMID- 11051605 TI - Generational differences in selenium status of women. AB - In this cross-sectional study of three generations of women, daughters (19-26 yr), mothers (40-58 yr) and maternal grandmothers (67-84 yr) from the same 10 families in central Ohio were studied to determine the effect of life-cycle differences, including estrogen status, on selenium status. Plasma and red blood cell (RBC) selenium and glutathione peroxidase (GPx) activities were determined and typical dietary selenium intakes were calculated from food-frequency questionnaires. Selenium status was lowest in the oldest generation. Plasma selenium of daughters and grandmothers were significantly lower than those of mothers, and plasma GPx and RBC selenium of grandmothers were also lower than those of the mothers. A positive correlation (r = 0.42, p < 0.04) was found between plasma estrogen and plasma selenium concentrations. Selenium intakes of all groups were adequate and no differences in selenium intakes were found among groups. The results of this study indicate that selenium status fluctuates during the female life cycle and is related to estrogen status. PMID- 11051606 TI - Microcalorimetric study of Escherichia coli growth inhibited by the selenomorpholine complexes. AB - The inhibitory or antibiotic action of four kinds of the selenomorpholine complex on a strain of Escherichia coli was studied by microcalorimetry. Differences in their capacities to inhibit the metabolism of this bacterium were observed. The extent and duration of the inhibitory effect on the metabolism as judged from the rate constant, k, and the half-inhibitory concentration, IC50, varied with the different drugs. The rate constant (k) of Escherichia coli (in the log phase) in the presence of the drugs decreased with increasing concentrations of the drugs (C). The relationship of k and C is nearly linear for (1) selenomorpholine and (2) selenomorpholine hydrochloride, but for (3) N,N'-methylene bisselenomorpholine and (4) N-dodecyl selenomorpholine, it is not linear. The experimental results reveal that the sequence of antibiotic activity of selenomorpholines is (3) and (4) > (1) > (2). PMID- 11051607 TI - Detection of potentially myocardial infarction susceptible individuals in indian population: a mathematical model based on copper and zinc status. AB - Zinc (Zn) and copper (Cu) concentrations in hair and urine of patients diagnosed and hospitalized for myocardial infarction (MI patients) and in their descendants (MI descendants) were estimated and compared with their age-matched healthy volunteers with no family history of MI (control group and control descendants). The data revealed approximately twofold higher Zn and twofold lower Cu in the urine of the patients; Zn was lower and Cu was higher in the urine of MI descendants than those of the patients (p < 0.001), but Zn in hair and urine was higher and Cu in hair was lower in MI descendants compared with their control counterparts (p < 0.001). The data suggested that there was a consistent rise in Zn and fall in Cu reserves in the genetically predisposed subjects (MI descendants) prior to the manifestation of clinical symptoms. Based on this, the data were subjected to logistic regression and a model was obtained to predict the susceptibility to MI (LR-MI), having impact factors values as follows: constant (C), -3.342; impact factor of body mass index, -0.776; impact factor of hair Zn, -2.449; impact factor of urine Zn, +3.441; impact factor of hair Cu, 15.077; impact factor of urine Cu, -24.153. For the equation Y = e(x)/(1 + e(x)), the value of x was obtained as follows: -3.342 + [BMI (kg/m2) (-0.776)] + [Hair Zn (micromol/g) (-2.449)] + [Urine Zn (micromol/L) (3.441)] + [Hair Cu (micromol/g) (-15.077)] + [Urine Cu (micromol/L) (-24.153)]. On substituting the values of BMI, hair Zn, urine Zn, hair Cu, and urine Cu in x, the response variable Y as zero for healthy controls and 0.99 or 99.9% susceptibility in MI patients were obtained. In between these two extremes, the response variable ranged between 0 and 0.99 or 99.9% susceptibility to MI in their descendants. It is envisaged that the MI patients have an operational component of a genetic disorder of ionic imbalance at a young age that can be exploited in making a prediction of susceptibility to heart stroke in individuals much before its onset and diagnosis in asymptomatic patients, particularly in genetic and epidemiological studies of MI. PMID- 11051608 TI - Determination of metal concentrations in animal hair by the ICP method: comparison of various washing procedures. AB - Concentrations of minor and trace elements (Mg, Ca, Fe, Ba, Cu, Zn, Cd, Ni, Al, Mn, Cr, Ti, and V) in the hair of three races of dogs (fox terrier, schnauzer, and mini schnauzer) were analyzed by the inductively coupled plasma-atomic emission spectrometry method. The influence of five washing solutions, deionized water, acetone, methanol, EDTA, and Triton X-100, on the concentrations measured in hair was investigated. Triton X-100 was found to be suitable to use for the removal of exogenous elements in multielemental hair analysis. Additionally, the results indicated that the concentration of the elements measured in the dogs' hair were similar to those reported for human hair. The relation between the element content in the dog hair and its color were similar to those found for human hair. PMID- 11051609 TI - Determination of thirty-two elements in human autopsy tissue. AB - Inductively coupled plasma-mass spectrometry (ICP-MS) was used to determine the concentrations of 32 elements in the human liver and kidney and 20 elements in the bone, obtained from 70 autopsied dead individuals (54 males, 16 females) between 18 and 76 yr of age from the North Bohemia territory of the Czech Republic. Geometric means, median, minimal-maximal range, as well as distribution and correlation analysis were calculated. Some significant differences among tissue concentrations of trace elements of the women and men were found. In the liver, medians of the concentrations of some elements were higher for men than that for women (Al: 770 vs 610 microg/kg; As: 42 vs 27 microg/kg; Cd: 1800 vs 1390 microg/kg; Rb: 3955 vs 3210 microg/kg; V: 160 vs 105 microg/kg). On the contrary, the content of other elements for men was lower (Bi: 0.8 vs 3.2 microg/kg; Cr: 57 vs 72 microg/kg; Hg: 228 vs 325 microg/kg; Zn: 57.1 vs 68.5 mg/kg). In the kidney of men, there were higher contents of Al (360 vs 245 microg/kg) and Hg (135 vs 75 microg/kg) and lower contents of Zn (47.7 vs 59.7 mg/kg) and I (135 vs 220 microg/kg) than those of women. In the case of bone, the concentrations of Cu and Rb were higher for men (1410 microg Cu/kg and 405 microg Rb/kg, respectively) than for women (655 microg Cu/kg and 285 microg Rb/kg, respectively). On the contrary, the content of Mn was considerably lower for men (110 microg Mn/kg) than for women (215 microg Mn/kg). PMID- 11051610 TI - Effect of lanthanum on aged seed germination of rice. AB - Attempts were made to promote germination of natural aged rice seeds by treating them with lanthanum nitrate. In tests to measure the germination rate, germination index, and vigor index of natural aged rice seeds were found to be significantly increased by lanthanum. It is treating aged rice seed with lanthanum nitrate that enhanced the respiratory rate and activities of superoxide dismutase, catalase, and peroxidase, and decreased superoxide O2*- and malondidehyde contents, and therefore reduced plasma membrane permeability. It suggests that lanthanum may be used to pretreat seed before sowing. PMID- 11051611 TI - Lanthanides enhance pulmonary absorption of insulin. AB - In an effort to investigate the enhancement effect of lanthanide ions (Ln3+) on the absorption of larger molecules from the pulmonary pathway, insulin (mol. wt. = 5730) was chosen as a model peptide. The absorption of insulin preadministered or coadministered with Ln3+ from the lung was investigated by means of an in situ pulmonary absorption experiment. The enhancement absorption of insulin by Ln3+ ions was evaluated by calculating the various bioavailabilities (Fr) of insulin from pulmonary absorption. Moreover, the temporal change of Gd content in serum was also investigated. Results showed that the promoting effect of Ln3+ on the bioavailability of insulin is closely related to its species, concentration, and delivery order. The effect of the median Ln3+ series was remarkably greater than that of light and heavy Ln3+. The anionic form of Gadolinium (Fr = 68.4%) seemed to be more effective compared with its cationic form (Fr = 59.5%). Coadministration of Gd3+ with insulin (Fr = 80.1%) was the most effective in increasing insulin absorption from the lung. Gd3+ was rapidly absorbed and metabolized to a normal level after 4 h. It was suggested that lanthanides in a very low concentration might become potent absorption enhancers to improve absorption of larger molecules via the pulmonary pathway. PMID- 11051612 TI - Vanadium-contaminated perioperatively administered infusion solutions and drugs. AB - Vanadium was determined in 51 solutions and drugs for intravenous administration and in 6 salt components of a multitrace element solution using electrothermal atomic absorption spectrometry. The highest V contaminations were found in albumin solutions showing values of more than 600 microg/L V. Two heparines contained 14.3 and 122 microg/L V, respectively. In the ultratrace element solution, 14.8 microg/L vanadium were determined. The most contaminated salt of this solution was Mn(II)-DL-aspartate (12.8 microg V/g). The concentrations of unbound V (Vf) in the albumin solutions were between 0.31 and 299 microg/L. The manufacturing process is the reason for V contamination. The biological half-life of V administered intravenously by albumin solutions in man was about 125 h. PMID- 11051613 TI - Effects of uranium poisoning on cultured preimplantation embryos. AB - The toxic effect of uranium in cultured preimplantation embryos of the mouse is presented. Embryos were obtained from hybrid females CBA x C57 BL following induction of superovulation and were incubated in M16 cultured medium. Two different experiments were performed. In one, embryos in a one-cell stage were placed in culture media with final concentrations of uranyl nitrate of 104 and 208 microg/mL during 120 h in the same dish. In the other experiment, embryos in a one-cell stage were placed in culture medium with uranyl nitrate with final U concentrations of 26, 52, 104, and 208 microg/mL. At 24 h, those embryos which had reached the two-cell stage were transferred to another culture dish to which fresh solutions with uranyl nitrate were added. The percentage of embryos in two cell stage, morula, early blastocyst, expanded blastocyst, and hatched blastocyst were recorded at 24, 72, 96 and 120 h of culture. The results obtained showed that concentrations as from 26 microg U/mL induced the delay of embryo development and the impairment of blastomere proliferation. The toxic effect of uranium increased in those experiments in which the embryos were transferred to a new medium. This embryo-culture system appears to be appropriate to evaluate the toxic effect of uranium on embryos removed from maternal influences and represents a suitable test system for environmental pollutants. PMID- 11051614 TI - Histochemical characterization of the lead intranuclear inclusion bodies. AB - A battery of histological and histochemical techniques was applied on the lead intranuclear bodies that have resuted in the kidneys of adult Wistar male rats receiving lead acetate in their diet to determine their nature. The intranuclear inclusion bodies have stained strongly with xanthene, anthraquinone, and trisulfonated basophilic dyes and weakly with dyes containing both positive and negative radicals, and they have responded negatively to acidophilic cationic dyes. They have also reacted positively to proteins and lead stains, but weakly to lipid stains, and negatively to Feulgen and methyl green pyronin techniques. The intranuclear bodies proved to be lead lipoprotein complexes containing sulfyhydryl groups and are basic in nature with orthochromatic, eosinophilic, argyrophilic, osmophilic, fuchsinophilic, and sudanophilic characteristics. PMID- 11051615 TI - Concentrations of trace elements in osteoarthritic knee-joint effusions. AB - Concentrations of the 18 elements, barium (Ba), beryllium (Be), bismuth (Bi), calcium (Ca), cadmium (Cd), cesium (Cs), copper (Cu), lanthanum (La), lithium (Li), magnesium (Mg), molybdenum (Mo), lead (Pb), rubidium (Rb), antimony (Sb), tin (Sn), strontium (Sr), thallium (Tl), and zinc (Zn), were determined in the synovial fluids of osteoarthritic knee joints and in the corresponding sera of 16 patients by inductively coupled plasma-mass spectrometry. Knee-joint effusions have lower elemental concentrations than their corresponding sera. For the essential elements Ca, Cu, Mg, and Zn and for the nonessential and toxic elements Ba, Be, Bi, La, and Sb, this difference was highly significant. Strong positive correlations between concentrations in effusions and sera for the essential elements Cu and Mg and for the nonessential elements Cs, Li, Rb, and Sr could be established. The grade of localized hyperperfusion of the knee region in the blood pool phase of 99mTc HDP bone scan indicating inflammation did not correlate with any elemental concentration determined. PMID- 11051616 TI - Population growth responses of Tetrahymena shanghaiensis in exposure to rare earth elements. AB - This article presents the population growth responses of Tetrahymena shanghaiensis s1 in exposure to rare earth elements (REEs). Both the light REEs (La, Sm) and the heavy REEs (Y, Gd) were investigated with 24- and 96-hr population growth assays to evaluate their aquatic toxicity. Four end points, cell count, frequency of neutral red (NR) uptake, total protein, and nucleic acid content were employed in the 24-h assay, and a population growth curve was plotted in the 96-h assay. The results of 24- and 96-h assays suggest a dual effect of REEs on T. shanghaiensis of the stimulated growth at low concentrations and the toxicity at higher ones. In the promoted growth of T. shanghaiensis, however, the cell density increased with an increased growth rate and the protein and nucleic acid content per 10(4) cells undertook no remarkable changes, which suggests possible cell growth control by REE. PMID- 11051617 TI - Pathogenesis of urinary tract infections: an update. AB - The pathogenesis of uncomplicated urinary tract infection (UTI) is complex and influenced by many host biological and behavioural factors and by properties of the infecting uropathogens. Most uncomplicated UTIs in women are not associated with underlying functional or anatomical abnormalities of the urinary tract, whereas sexual intercourse, spermicide use, a history of recurrent UTI and recent antimicrobial chemotherapy are important risk factors. A maternal history of UTI and young age at first UTI, as well as sexual intercourse and spermicide use, are risk factors for recurrent UTI in young women. In some young healthy women, especially those with 'low UTI risk' behaviour, features of pelvic anatomy appear to be associated with UTI risk. In postmenopausal women, anatomical and functional characteristics of the genitourinary tract are more strongly associated with UTI risk than in younger women. A genetic predisposition to recurrent UTI is suggested by the association of recurrent UTI in certain age groups with the ABH blood group non-secretor phenotype, a maternal history of UTI and early age at onset of UTI. Virulence determinants of uropathogens are much more important in the normal host than in the host who has a functional or anatomical abnormality of the genitourinary tract. PMID- 11051618 TI - Comparative potency of mecillinam and other beta-lactam antibiotics against Escherichia coli strains producing different beta-lactamases. AB - The activity of mecillinam, a beta-lactam antibiotic with high affinity for gram negative penicillin-binding protein 2 (PBP2), was assessed against ampicillin resistant Escherichia coli strains producing beta-lactamases representing the three molecular classes, A (TEM-1 and -3, SHV-3 and IRT-5), C (AmpC) and D (OXA 3). The antimicrobial activity of mecillinam and other beta-lactam antibiotics was evaluated by determining their MICs on Mueller-Hinton agar. The time course of hydrolysis in crude extracts prepared from the various beta-lactamase producing strains was also measured and was used to determine the relative rate of hydrolysis and the apparent affinity for ampicillin, cephalothin and mecillinam. When compared with the other beta-lactam antibiotics, mecillinam demonstrated significantly greater antibacterial potency and higher stability to beta-lactamase hydrolysis in TEM-, IRT- and AmpC-producing isolates. These findings confirm that the antimicrobial potency of mecillinam compares favourably with those of the other penicillins included in the present study, suggesting that mecillinam use in the treatment of infections caused by gram-negative bacteria should be re-evaluated. PMID- 11051619 TI - The ECO.SENS Project: a prospective, multinational, multicentre epidemiological survey of the prevalence and antimicrobial susceptibility of urinary tract pathogens--interim report. AB - The ECO.SENS Project is the first international survey to investigate the prevalence and susceptibility of pathogens causing community-acquired, uncomplicated urinary tract infections (UTIs) in women. At 240 centres in 17 countries, female patients presenting with symptoms of uncomplicated UTIs were asked to provide a urine sample for testing for the presence of leucocytes and bacteria. The bacteria were identified and their susceptibility to 12 antibiotics commonly used in the treatment of UTIs was determined. The objective of the survey was to collect 5000 urine samples to obtain approximately 3500 isolates of defined uropathogens. This interim report includes the results from 1960 urine samples, 75% of which contained a uropathogen. Escherichia coli accounted for the majority (80%) of uropathogens isolated in all 17 countries. The rates of resistance among E. coli strains were: ampicillin and sulphamethoxazole, 30%; trimethoprim alone or with sulphamethoxazole, 15%; nalidixic acid, 6%; ciprofloxacin, 3%; amoxycillin-clavulanic acid, mecillinam, cefadroxil, nitrofurantoin and fosfomycin, < or =2%. The use of ampicillin, sulphonamides and trimethoprim alone or with sulphamethoxazole needs to be reconsidered. The seemingly rapid increase in quinolone resistance among community-acquired E. coli in some of the countries gives cause for concern. PMID- 11051620 TI - Treatment options for acute uncomplicated cystitis in adults. AB - Urinary tract infection (UTI) is classified as uncomplicated if it occurs in a patient with a structurally and functionally normal urinary tract. Acute uncomplicated cystitis is observed chiefly in women. It needs, however, to be differentiated depending on whether it occurs in premenopausal, postmenopausal or pregnant women. Only a small number of 15-50 year old, otherwise healthy men suffer acute uncomplicated cystitis. In premenopausal, non-pregnant women, single dose antimicrobial therapy is generally less effective than the same antibiotic used for longer duration. However, most antimicrobial agents given for 3 days are as effective as those given for longer duration, and adverse events tend to be found more often with longer treatment. Trimethoprim (or co-trimoxazole) can be recommended as first-line empirical therapy only in communities with resistance rates of uropathogens to trimethoprim of < or =10-20%. Otherwise fluoroquinolones are recommended. Alternatives are fosfomycin trometamol or beta-lactams, such as second- or third-generation oral cephalosporins or pivmecillinam, especially when fluoroquinolones are contraindicated or a high proportion (>10%) of Escherichia coil strains in the community are already resistant to fluoroquinolones, as in Spain, for example. Recurrent UTIs are common among young, healthy women even though they generally have anatomically and physiologically normal urinary tracts. The following prophylactic antimicrobial regimens are recommended: (i) the use of long-term, low-dose prophylactic antimicrobials taken at bedtime; (ii) post-coital prophylaxis for women in whom episodes of infection are associated with sexual intercourse. Other prophylactic methods are not as yet as effective as antimicrobial prophylaxis. PMID- 11051621 TI - Which antibiotics are appropriate for treating bacteriuria in pregnancy? AB - Bacteriuria in pregnancy, with or without clinical symptoms, is frequent. If left untreated, it can in 20-30% of cases lead to acute pyelonephritis, which is a serious threat to the mother and fetus, increasing the risk of preterm labour and low birthweight infants. This paper is a review of the literature concerning antibacterial treatment of bacteriuria in pregnancy. It is crucial to ensure that drugs to be used in pregnancy are safe and effective. Established first-line drugs such as ampicillin (pivampicillin) and amoxycillin, and other commonly used treatments such as trimethoprim-sulphamethoxazole, are associated with a high degree of resistance in Escherichia coli, the most common pathogen in the urinary tract. A recent survey of physicians in Denmark, Finland, Norway and Sweden confirms that beta-lactam antibiotics (particularly pivmecillinam) and nitrofurantoin are the drugs of first choice in the treatment of bacteriuria in pregnancy in the Nordic countries. No teratogenic effects have been associated with these agents. In contrast to nitrofurantoin, pivmecillinam is also efficient against pyelonephritis. In spite of resistance in E. coli and possible adverse effects on the fetus, many physicians still prescribe sulphonamides during the first two trimesters of pregnancy. PMID- 11051622 TI - Pivmecillinam in the treatment of urinary tract infections. AB - The efficacy of pivmecillinam for empirical treatment of acute uncomplicated urinary tract infection (UTI) was initially reported in clinical trials published in the 1970s and 1980s. Bacteriological cure rates observed in these trials were consistently >85%, and studies of different dosing regimens suggested that a 3 day course was appropriate. Comparative studies reported that pivmecillinam was equivalent to other antimicrobial agents in terms of clinical and bacteriological outcomes. These studies also documented that pivmecillinam was effective for treatment of Staphylococcus saprophyticus infections, was acceptable for use in pregnancy and was well tolerated. Subsequent widespread use of pivmecillinam in Scandinavian countries has led to a body of clinical experience which confirms the efficacy and safety of this antimicrobial agent in the treatment of acute cystitis. Recently, two large, prospective, randomized, double-blind, multi centre clinical trials have been completed to assist in defining the role of this antimicrobial agent in the treatment of acute cystitis. A comparison of 3 day courses of pivmecillinam or norfloxacin, both at 400 mg bd, showed higher bacteriological cure rates with norfloxacin but generally similar clinical outcomes. A second, dose-ranging study found that pivmecillinam, given bd for 7 days, led to superior bacteriological and clinical outcomes at short-term follow up than the 3 day regimen. Pooling bacteriological outcomes from the two studies showed similar outcomes with 7 days of pivmecillinam 200 mg bd or 3 days of norfloxacin 400 mg bd. The shorter, 3 day, course achieved similar short-term clinical outcomes to 7 days of pivmecillinam and 3 days of norfloxacin in women aged < or =50 years. These recent studies confirm earlier reports and clinical experience that pivmecillinam is effective and well tolerated for the treatment of acute cystitis in women. PMID- 11051623 TI - Effect on the human normal microflora of oral antibiotics for treatment of urinary tract infections. AB - Oral administration of antibiotics for treatment of urinary tract infections (UTIs) can cause ecological disturbances in the normal intestinal microflora. Poorly absorbed drugs can reach the colon in active form, suppress susceptible microorganisms and disturb the ecological balance. Suppression of the normal microflora may lead to reduced colonization resistance with subsequent overgrowth of pre-existing, naturally resistant microorganisms, such as yeasts and Clostridium difficile. New colonization by resistant potential pathogens may also occur and may spread within the body or to other patients and cause severe infections. It is therefore important to learn more about the ecological effects of antibacterial agents on the human microflora. The impact on intestinal microorganisms of oral antibiotics used for the treatment of UTIs is reviewed here. Ampicillin, amoxycillin and co-amoxiclav suppress both the aerobic and anaerobic intestinal microflora with overgrowth of ampicillin-resistant Enterobacteriaceae. Pivmecillinam also affects the intestinal microflora, suppressing Escherichia coli, but does not have a major effect on the anaerobic microflora. Several orally administered cephalosporins, such as cefixime, cefpodoxime, cefprozil and ceftibuten, reduce the number of Enterobacteriaceae and increase the number of enterococci. Colonization with C. difficile has also been observed. Fluoroquinolones eliminate or strongly suppress intestinal Enterobacteriaceae, but affect enterococci and anaerobic bacteria only slightly. When antimicrobial agents are prescribed for the treatment of UTIs, not only the antimicrobial spectrum of the agent but also the potential ecological disturbances, including the risk of emergence of resistant strains, should be considered. PMID- 11051624 TI - Survey on antibiotic usage in the treatment of urinary tract infections. AB - Ninety-two clinical microbiologists, infectious disease clinicians and clinicians with an interest in the management of urinary tract infections (UTIs) participated in an interactive session concerning the management of acute uncomplicated lower UTI. The antibacterials considered most appropriate as first line agents were trimethoprim, co-trimoxazole, pivmecillinam, nitrofurantoin and fluoroquinolones. The current level of usage of fluoroquinolones for lower UTIs was, however, considered inappropriate by most delegates from a 'societal perspective', in terms of spread of resistance and potential impact on the environment. PMID- 11051625 TI - Environmental risk assessment of antibiotics: comparison of mecillinam, trimethoprim and ciprofloxacin. AB - The effects of mecillinam, trimethoprim and ciprofloxacin, antibiotics used in the treatment of urinary tract infections, on the aquatic environment were assessed. Mecillinam and ciprofloxacin were both readily biodegradable (primary degradation) in activated sludge, whereas trimethoprim persisted. The toxicity of these antibiotics towards sludge bacteria, a green alga, a cyanobacterium, a crustacean and a fish were investigated; both mecillinam and ciprofloxacin were highly toxic to the cyanobacterium Microcystis aeruginosa (EC50 in the range 5-60 microg/L). Risk characterization for the aquatic environment was performed for the three compounds by calculating the predicted environmental concentration (PEC) and the predicted no-effects concentration (PNEC). A PEC/PNEC ratio of <1 indicates that, with the present pattern of use, no environmental risk is expected. PEC/PNEC ratios of <1 for present usage in Europe were found for mecillinam and trimethoprim whereas a PEC/PNEC ratio >1 was found for ciprofloxacin. PMID- 11051626 TI - Ecological antibiotic policy. AB - Development of resistance to antibiotics is a major problem worldwide. The normal oropharyngeal flora, the intestinal flora and the skin flora play important roles in this development. Within a few days after the onset of antibiotic therapy, resistant Escherichia coli, Haemophilus influenzae and Staphylococcus epidermidis can be detected in the normal flora of volunteers or patients. Horizontal spread of the resistance genes to other species, e.g. Salmonella spp., Staphylococcus aureus and Streptococcus pneumoniae, occurs by conjugation or transformation. An ecologically sound antibiotic policy favours the use of antibiotics with little or no impact on the normal flora. Prodrug antibiotics which are not active against the bacteria in the mouth and the intestine (before absorption) and which are not excreted to a significant degree via the intestine, saliva or skin are therefore preferred. Prodrugs such as pivampicillin, bacampicillin, pivmecillinam and cefuroxime axetil are favourable from an ecological point of view. Experience from Scandinavia supports this, since resistance to mecillinam after 20 years of use is low (about 5%) and stable. PMID- 11051627 TI - XIIIth International AIDS Conference: new thoughts on old drugs. PMID- 11051628 TI - Case report: one-and-a-half-syndrome and tuberculosis of the pons in a patient with AIDS. AB - A patient with advanced AIDS presented with right conjugate gaze palsy and impaired adduction on left gaze (the one-and-a-half syndrome). The responsible pontine lesion involved the ipsilateral abducens nucleus and the adjacent medial longitudinal fasiculus (MLF), as demonstrated by magnetic resonance imaging (MRI). Tuberculosis (TB) was the etiology of the brainstem lesion. The patient had complete recovery after anti-tuberculosis treatment. The nuclear location of the tuberculoma is unusual and the reversal of this neuro-ophthalmologic syndrome is noteworthy. PMID- 11051629 TI - Salvage therapy for HIV infection: when and how. PMID- 11051630 TI - Physician beliefs about antiretroviral adherence communication. AB - Adherence to antiretroviral regimens is thought to be imperative for HIV-positive patients. However, little is known about health care professionals' willingness to discuss adherence with patients, nor what barriers, if any, may impede their adherence communication. The aims of the study were to examine physicians' beliefs regarding: (1) the efficacy of adherence communication, (2) the barriers which impede such communication, and (3) their roles and responsibilities regarding adherence communication. Semi-structured interviews were conducted with 15 physicians who specialize in providing care to HIV-positive patients. Results showed that most physicians believed that communicating with patients about adherence issues was important, even though doing so may not have an immediate impact on patients' behaviors. Physicians believed that adherence communication was a good use of their time and fell within their role as a primary care professional. However, time constraints were cited as a barrier to performing adherence communication, indicating that physicians may do less communicating about adherence than may be optimal if they are burdened with other tasks. Results highlight the need to develop innovative strategies, such as the use of multi-disciplinary adherence teams, to ensure that each HIV-positive patient receives the optimal amount of information about and support for adherence. PMID- 11051631 TI - Public disclosure of a child's HIV infection: impact on children and families. AB - As an increasing number of children infected with HIV live to older ages, the question of disclosure of the diagnosis (to the child and to others) becomes more crucial. Disclosure of a child's HIV diagnosis is a controversial and emotionally laden issue. One reason that families avoid disclosure is their fear of the negative impact on the child and family that the stigma associated with AIDS can bring. At the other end of the spectrum, are those families that choose to publicly disclose an HIV diagnosis. There are a number of reasons that a family may choose to go public with their child's diagnosis, although this has never been systematically assessed. The dearth of literature and research regarding the emotional impact of public life on a child as well as the interest of a number of non-public HIV-infected children to "go public" served as an impetus to conduct a study that directly examined the impact public disclosure has on the HIV-infected child and family. Specifically, findings pertaining to the decision making process, the impact public disclosure has had on the child's family, and the child's sense of self-worth at the time of the study and then again 4 years later are reviewed. Findings and implications for future research as well as interventions and strategies aimed at counseling families considering "going public" and helping to normalize the public child's life are discussed. PMID- 11051632 TI - amfAR's World AIDS Day symposium, United Nations, November 30, 1999. Introduction and greeting. PMID- 11051633 TI - amfAR's World AIDS Day symposium, United Nations, November 30, 1999. The health and wealth of nations. PMID- 11051634 TI - Converging agendas: AIDS and business. PMID- 11051635 TI - A moment in time: AIDS and business. AB - Business has transformed the planet. But this gives it new responsibilities. People now expect business leaders to lead--and not just respond when things go wrong. HIV/AIDS is a global problem, with over 16.3 million people now thought to have died of the disease (Global Summary of HIV/AIDS Epidemic, UNAIDS, December 1999). Without action now, the pandemic will worsen, health services will come under relentless pressure and the number of people dying will increase exponentially. So why should business sit up and take notice? First: money. AIDS is slowly strangling many businesses and economies--and in a global market, everyone eventually suffers. Without profit, there is no business--so the business community needs to act to protect its bottom line. Second: people. Over 80% of those dying are in their 20s, 30s, and 40s. Businesses are losing workers and customers, and human networks that have taken decades to build. Third: imagination. Business is inventive, creative and fast-moving. It has the opportunity to use these strengths for the benefit of the wider community. It's time to pit business ideas (and some money, too) against the threat of AIDS. The course of the AIDS epidemic is not inevitable. The world's businesses have the skills and intensity to make a measurable difference, especially if they find public sector and NGO partners with whom they share a vision. A focused, coordinated, results-driven effort will hit AIDS hard. The HIV virus moves fast (and is mutating all the time). Business has the opportunity to make a difference. It must grasp this opportunity. And grasp if fast. PMID- 11051636 TI - Integrase inhibitor studied. PMID- 11051637 TI - Approval sought for new PI. PMID- 11051638 TI - Warning for oral solution. PMID- 11051639 TI - Blood substitute being developed. PMID- 11051640 TI - Syphilis programs under way. PMID- 11051642 TI - [Variability of life and medicine]. PMID- 11051641 TI - New drugs fight TB. PMID- 11051643 TI - [Physiopathology and treatment of hypersensitivity pneumonitis]. PMID- 11051644 TI - [Etiology and clinical presentation of cholesterol cholelithiasis]. PMID- 11051645 TI - [Hypercalcemia in malignant tumor--with special reference to ATL]. PMID- 11051646 TI - [Cardiac failure and neurohumoral factors]. PMID- 11051647 TI - [Study of complements for clinicians--the basis of host defense system and an overview for the 21st century]. PMID- 11051648 TI - [Genes and hypertension]. PMID- 11051649 TI - [Genetic engineering study in hypertensive research]. PMID- 11051650 TI - [Central nervous system and hypertension]. PMID- 11051651 TI - [Kidney and hypertension]. PMID- 11051652 TI - [The renin-angiotensin system and hypertension--the summary of the 20th century and expectation on the 21st century]. PMID- 11051653 TI - [Diffuse lung diseases]. PMID- 11051654 TI - [Obstructive respiratory diseases]. PMID- 11051655 TI - [Allergic lung diseases]. PMID- 11051656 TI - [Molecular physiology and clinical manifestation of lung cancer]. PMID- 11051658 TI - [The current status of hepatitis viruses]. PMID- 11051657 TI - [Respiratory tract infections]. PMID- 11051659 TI - [Topics on acute hepatitis and fulminant hepatitis]. PMID- 11051660 TI - [Topics on type B chronic hepatitis]. PMID- 11051661 TI - [Topics on type C chronic hepatitis]. PMID- 11051662 TI - [Myocardial, renal, and vascular lesions caused by hepatitis viruses]. PMID- 11051663 TI - [Lesions of the skin, mucous membrane, muscles, and hematopoietic organs caused by hepatitis viruses]. PMID- 11051664 TI - [Legislative viewpoint--evaluation and the future of the law concerning organ transplantation]. PMID- 11051665 TI - [Current status and the future of organ transplantation. 1) Kidney and pancreas transplantation]. PMID- 11051666 TI - [Current status and the future of organ transplantation. 2) On heart and heart lung transplantation]. PMID- 11051667 TI - [Current status and future of organ transplantation. 3) Lung transplantation]. PMID- 11051669 TI - [Approach to organ transplantation and problems at organ providing organizations]. PMID- 11051668 TI - [Problems in encouraging multiple organ procurement from subjects who succumbed to brain death in Japan]. PMID- 11051670 TI - [Current status of heart failure therapy]. PMID- 11051671 TI - [Diagnosis and treatment of primary biliary cirrhosis of the liver]. PMID- 11051672 TI - [Mycoses]. PMID- 11051673 TI - [Life-saving emergency care of patients with heart diseases: prehospital emergency care that determines life or death of patients]. PMID- 11051674 TI - [New strategy for the management of pneumonia of the aged]. PMID- 11051675 TI - [Clinical aspect of IgA nephropathy]. PMID- 11051676 TI - [Evaluation of the serum pepsinogen level as a method to screen stomach cancer]. PMID- 11051677 TI - [Sjogren's syndrome and lymphoproliferative lesions]. PMID- 11051678 TI - [Treatment of hypertension in diabetic patients]. PMID- 11051679 TI - [Primary care and tuberculosis]. PMID- 11051680 TI - [Progress in studies of Parkinson's disease]. PMID- 11051681 TI - [The brain structure related to the language function]. PMID- 11051682 TI - [Diagnosis and treatment of acute myocardial infarction (AMI)]. PMID- 11051683 TI - [Clinical aspect of autoimmune hemolytic anemia]. PMID- 11051684 TI - [Physiopathology and treatment of chronic pancreatitis]. PMID- 11051685 TI - [Physiopathology and treatment of focal segmental glomerulosclerosis]. PMID- 11051686 TI - [Clinical aspect of growth hormone deficiency in adults]. PMID- 11051687 TI - [Clinical aspect of thrombotic diathesis]. PMID- 11051688 TI - [Evidence-based surgery: from concept to everyday practice]. PMID- 11051689 TI - [Surgical treatment of pancreatic cancer: curative resections]. AB - Curative resection of pancreatic adenocarcinoma can only be performed in 10% of patients. This review article reports resectability rates and criteria, results of pancreatic resection and prognostic factors. Lymph node and/or vascular involvement and retroperitoneal tissue invasion constitute very poor prognostic factors; however, lymph node involvement limited to the first draining nodes and limited invasion of the mesenteric-portal vein do not constitute contraindications to surgical resection. Cephalic pancreaticoduodenectomy is still the reference procedure and its postoperative mortality has greatly decreased. The risk of pancreatic fistula mainly depends on the friability of the pancreatic stump. Median survival rate after tumour resection is usually limited between 12 and 18 months. Five-year actuarial survival rate is no more than 5%, but after curative resection (RO), it may be as high as 20 to 25% in recent surgical series. Concomitant or neoadjuvant chemotherapy-radiotherapy, currently under evaluation, may increase resection and survival rates. PMID- 11051690 TI - [Conservative treatment of adenocarcinomas of the anorectal junction by preoperative radiotherapy and intersphincteral resection]. AB - AIM: Adenocarcinomas of the anorectal junction, especially T3 lesions, are usually treated by abdominoperineal resection. The aim of this study was to evaluate oncologic and functional results following conservative radiosurgical treatment of cancers of the anorectal junction. METHODS: From 1990 to 1999, among 395 patients with rectal carcinoma, 31 had sphincter-saving resection for a tumour located between 2 to 4.5 cm (mean 3.6) from the anal verge. There were 16 men and 15 women, mean age 62 years (range 30-86). There were 5 T2, 23 T3 and 3 T4 tumours; 17 were N1 and 3 were M1. Preoperative radiotherapy was performed in 26 patients (dose: 46 Gy, range: 36-54), with concomitant chemotherapy in 14 cases. Intersphincteric resection was performed six weeks after neoadjuvant treatment. Coloanal anastomoses were associated with a colonic pouch in 22 cases and with a protecting stoma in all cases. RESULTS: There was no postoperative mortality. Seven complications occurred: 3 anastomotic fistulas, 3 pelvic haemorrhages and 1 acute pancreatitis. Three patients had a definitive stoma. After preoperative radiotherapy, down-staging (pT0-2 N0) occurred in 46% of cases (12/26). Distal margin was 2.2 cm (range: 1-3) and was microscopically safe in all cases. Lateral margin was safe (> or = 1 mm) in 97% of cases. With a mean follow-up of 36 months, no local recurrence was suspected. Twenty-six patients (84%) were alive, 23 free of disease. Half of the patients had perfect continence, whereas the other half had occasional minor soiling. Functional results were better in patients with a colonic pouch. CONCLUSION: Conservative treatment of carcinomas of the anorectal junction is possible without compromising pelvic control and patient survival. Pelvic control was probably achieved by using preoperative radiotherapy with intersphincteric resection, ensuring safe distal and lateral margins. PMID- 11051691 TI - [Prognostic factors in advanced pancreatic cancer. Multivariate analysis of predictive survival score. University Surgery Association]. AB - STUDY AIM: To identify prognostic factors in advanced pancreatic cancer and to define a predictive score. PATIENTS AND METHOD: One hundred and sixty six patients were included in this multicentre study. Seventeen covariables were prospectively collected for each patient. Covariables associated with survival (p < 0.10) were analysed by a stepwise Cox model. RESULTS: Four prognostic factors were selected on multivariate analysis: pain (RR 1.5; CI: 1.1-2.0), ascites (RR 1.7; CI: 1.0-2.9), weight loss > 10 kg (RR 1.4; CI: 1.0-2.0), and metastases (RR 2.3: CI: 1.6-3.2). A score was defined by attributing a value of one for pain, ascites and weight loss, and two for metastases. Patients with a score > 2 had a median survival of 2 months (SE: 0.5), and patients with a score < or = 2 had a median survival of 6 months (SE: 0.6) (Logrank p < 0.0001). CONCLUSION: The proposed score may be helpful in therapeutic decisions concerning surgery, palliative chemotherapy and/or radiotherapy. However, this score must be validated on an independent series of patients. PMID- 11051692 TI - [Peritoneal carcinomatosis treated by cytoreductive surgery and intraperitoneal chemohyperthermia]. AB - STUDY AIM: The aim of this prospective non-randomized trial was to report a series of intraperitoneal carcinomatosis due to miscellaneous causes, treated by intraperitoneal hyperthermic perfusion (IPHP) and cytoreductive surgery. PATIENTS AND METHOD: From January 1995 to May 1999, 35 patients were treated by IPHP and 26 of them underwent maximal cytoreductive surgery. IPHP was performed for 60 minutes at an intraperitoneal temperature of 42 degrees C with Mitomycin C (10 mg/L) or cisplatinum (12 mg/L) at a flow rate of 0.9 L/min. RESULTS: There was one (2.8%) postoperative death due to respiratory complications on day 16. Three patients (8.5%) were admitted to the intensive care unit. A high morbidity rate (54%) was observed with intra-abdominal complications in 28.5% of patients, requiring reoperation in three patients. In patients with stages 1 and 2 peritoneal carcinomatosis (granulations less than 5 mm), the 12- and 24-month survival rates were 63.1% and 31.5%, respectively. In patients with advanced stage 3 (diffuse malignant nodules less than 2 cm) and stage 4 carcinomatosis (malignant nodules larger than 2 cm), the 12- and 24-month survival rates were 31.2% and 12%, respectively. Six patients survived for more than 30 months. CONCLUSION: IPHP appears to be an effective treatment for peritoneal carcinomatosis. IPHP combined with cytoreductive surgery is aggressive with a high morbidity rate. Rigorous patient selection is necessary. IPHP is still under evaluation. Prospective randomized trials with identical IPHP protocols are required. PMID- 11051693 TI - [Incidence of sphincter ruptures in anal incontinence: ultrasound study]. AB - STUDY AIM: To assess the incidence of endosonographic anal sphincter defects in a population of patients complaining of anal incontinence. PATIENTS AND METHODS: Seventy two consecutive patients (10 men, 62 women, mean age: 55 years) with anal incontinence underwent transanal sonography. In this series, 22 patients had a history of anal surgery (group I), while 50 patients (46 parous women, 4 men) had not been previously operated (group II). RESULTS: Fifty patients (69.4%) had an anal sphincter defect identified on transanal sonography: 6 isolated internal sphincter defects (12%), 18 isolated external sphincter defects (36%) and 26 combined sphincter defects (52%). The incidence of sphincter defects was similar in the surgical and medical group (81.8% vs 64%, p > 0.05). All but one of the isolated internal sphincter defects were observed in group I. Among the 46 parous women of group II, the use of forceps was not associated with a significantly higher frequency of anal sphincter defects (72% vs 64%, p > 0.05). CONCLUSION: This study confirms the high incidence of endosonographic anal sphincter defects in patients with anal incontinence. Isolated lesions of the internal sphincter are mainly seen after anal surgery. In our group of parous women, the use of forceps did not increase the incidence of anal sphincter lesions. PMID- 11051694 TI - [Abdominal aortic aneurysm and autosomal dominant polycystic kidney disease]. AB - AIM: The aim of this study was to report a series of patients with autosomal dominant polycystic kidney disease operated for abdominal aortic aneurysm. PATIENTS AND METHODS: From 1986 to 1999, seven patients with this pathologic association were operated for aneurysm by the same surgeon. All were males, 47 to 69 years old (mean: 57.7). All were hypertensive and heavy smokers. Four were treated by hemodialysis. In five patients, the aneurysm was an incidental discovery, while two patients presented signs of obstructive arterial disease of the lower limbs. Ultrasound was the routine screening investigation, completed by aortography in all patients and by computed tomography in 2 patients. Surgical treatment consisted of intrasaccular repair of the aneurysm with a straight aortic tube (n = 5), a bifurcated prosthesis from the aorta to both common iliac arteries (n = 1) and a bifurcated prosthesis from the aorta to both common femoral arteries (n = 1). RESULTS: There was no postoperative mortality or morbidity. Two late deaths (at 5 and 8 years) occurred from myocardial infarction. Only one patient subsequently received a kidney transplant. Repairs were verified by postoperative angiography: anatomical results were satisfactory in all patients. Only nine similar cases have been published in the literature, including two deaths from ruptured aneurysm. CONCLUSIONS: The clinical diagnosis of aortic aneurysm is difficult in patients with polycystic kidneys due to renal volume. Ultrasound scan of the aorta is recommended to screen these patients for aneurysm. The data of our series show that the main cause of aortic aneurysms is atheroma and that a pathogenic link between this lesion and polycystic kidney disease is questionable. Elective aortic repair is recommended in order to avoid rupture of the aneurysm. PMID- 11051695 TI - [Treatment of hydatid bilio-bronchial and bilio-pleuro-bronchial fistulas by thoracotomy]. AB - STUDY AIM: The aim of this study was to report the results of surgical treatment of hydatid bilio-bronchial et bilio-pleuro-bronchial fistulas via thoracotomy. MATERIAL AND METHODS: From 1990 to 1998, 63 cases were observed in the same center. Biliptysis was the main symptom in 72% of cases. The diagnosis was based on chest radiography and abdominal ultrasonography; both examinations visualised the cyst, intrathoracic collections, a diaphragmatic breach and biliary lesions. All patients were treated by one-stage thoracotomy. The procedures consisted of lung resection (lobectomy and/or segmentectomy) (n = 47) and decortication (n = 16) in the chest, cyst dome resection (n = 61) or partial pericystectomy (n = 12) in the abdomen and suture of the diaphragmatic defect in all cases after hepato diaphragmatic disconnection. An additional laparotomy was necessary in 4 cases. RESULTS: There were 8 deaths (12.7%): one intraoperative death due to haemorrhage and seven postoperative deaths, mostly related to pulmonary complications. Postoperative complications (14.3%) were mainly respiratory. Clinical and radiological results were good with a one-year follow-up. CONCLUSION: Bilio bronchial and bilio-pleurobronchial fistulas due to hydatid cyst are rare, but severe diseases. They are responsible for lesions at three levels: abdominal, diaphragmatic and thoracic. A high perioperative mortality rate was observed. Thoracotomy is the best approach for surgical treatment at all three levels. PMID- 11051696 TI - [Cephalic duodenopancreatectomy with preservation of pancreaticoduodenal arcades in coeliac trunk occlusion]. AB - In the case of complete occlusion of the coeliac trunk, the hepatic and splenic arterial blood supply is ensured by the superior mesenteric artery. Interruption of this collateral circulation by pancreaticoduodenectomy carries a risk of hepatic ischaemia and anastomotic leaks. In addition to the various techniques of coeliac trunk revascularization, preservation of the gastroduodenal artery and pancreaticoduodenal arcades may help to preserve the arterial blood supply. The authors report the case of a 58-year-old female with a benign tumour of the pancreatic head and partially corrected coeliac trunk occlusion. Pancreaticoduodenectomy was performed with preservation of the pancreaticoduodenal arcades and gastroduodenal artery. PMID- 11051697 TI - [Crohn's disease of the appendix]. AB - A giant appendix is an extremely rare and improbable finding during surgery in suspected cases of acute appendicitis. Although this condition is primarily suggestive of neoplasia, it is usually due to an inflammatory or infectious disease. We report a case of Crohn's disease limited to the appendix, which was diagnosed after a short right ileocolectomy. Only 156 similar diagnoses have been reported in the literature to date. This disease appears to have a benign course and therefore differs from classical Crohn's ileocolitis. For this reason, these patients do not require any specific investigation or follow-up. PMID- 11051698 TI - [Role of outpatient surgery in breast surgery. Prospective feasibility study]. AB - STUDY AIM: The objective of this prospective study was to assess the feasibility of outpatient breast surgery, the reasons for inpatient procedures (IPP), the reasons for conversion and the conversion rate, and the postoperative morbidity after outpatient procedures (OPP). PATIENTS AND METHODS: In 1999, among 625 patients eligible for OPP (diagnostic surgery or conservative curative surgery), OPP was performed in 418 patients (67%) and IPP was performed in 207 patients (33%). The reasons for IPP rather than OPP were environmental (64%) rather than medical (16%). RESULTS: The conversion rate to conventional surgery was 12.4% and the definitive OPP rate was 58.6%. The reasons for conversion were more often medical (50%) and environmental (21%) than surgical (23%). The morbidity, except for axillary seroma, was similar for OPP and IPP. The axillary seroma rate after axillary lymph node dissection was higher with OPP (27.4 vs 16.1%). CONCLUSION: OPP is a good alternative to IPP in breast surgery, especially for diagnostic purposes. OPP is also feasible for partial mastectomy with axillary lymph node dissection, but patients must be clearly informed about the risks of axillary morbidity. The patients' quality of life and satisfaction index should also be evaluated. PMID- 11051699 TI - Ageing and the health sector in Sri Lanka. PMID- 11051700 TI - Quality in medical education. PMID- 11051701 TI - The pattern of hypogammaglobulinaemia in Sri Lankan children. AB - OBJECTIVE: To evaluate the prevalence of hypogammaglobulinaemia in Sri Lankan children who present with recurrent or severe bacterial infections. DESIGN: A descriptive study. SETTING: Medical Research Institute (MRI), Colombo. SUBJECTS: 100 children between the ages of four months to twelve years referred to the Department of Immunology, MRI, for evaluation of immune status during four years from 1993 to 1997. MEASUREMENTS: Immunoglobulin G, A and M levels were measured using radial immunodiffusion technique. RESULTS: 22 out of 100 children had an underlying antibody deficiency, of whom IgA deficiency was the commonest (18 patients). Two patients had low IgG and A and elevated IgM levels, and they were diagnosed as having X linked-hyper-IgM syndrome. One patient had deficient IgA and IgM, and all three immunoglobulins were deficient in another. CONCLUSIONS: Results indicate that IgA deficiency is the commonest immunodeficiency in Sri Lanka, which is comparable with studies done in the West. This study also shows the need to improve the standard of care in patients with immunodeficiency. PMID- 11051702 TI - Feasibility of canine oral rabies vaccination in Sri Lanka--a preliminary report. AB - OBJECTIVE: To test the feasibility of introducing oral rabies vaccine (vaccinia recombinant rabies glycoprotein) to improve domestic dog immunisation coverage in a selected area. DESIGN: A prospective field trial. SETTING: Panadura, an area relatively isolated by waterways, making it a suitable field laboratory. METHODS: A routine parenteral vaccination program was carried out. A house to house survey identified the residual non-vaccinated dogs in the selected area. Oral vaccine was offered to the non-vaccinated domestic dogs. RESULTS: Of 4322 dogs in the households, 1242 dogs (28.7%) were eligible for oral vaccine. 659 (53%) were considered to have accepted the oral vaccine with release of the vaccine in the oral cavity. CONCLUSION: It is feasible to use oral rabies vaccine to enhance immunisation coverage. PMID- 11051703 TI - Use of antivenom serum in snake bite: a prospective study of hospital practice in the Gampaha district. AB - OBJECTIVE: To record current practices in hospital management of snake bite, especially with regard to use of antivenom serum (AVS). METHODS: Management of all snake bite victims admitted to the four main hospitals of the Gampaha district was prospectively studied during a 5-month period. A pretested data collection sheet was used. Relevant information was obtained from patients, accompanying persons, medical staff and hospital records. RESULTS: 466 patients (M:F = 7:3; 402 adults and 64 children) were admitted following snake bite during the study. The offending snake was identified in 357 (76.6%) cases [172 (36.9%) by examining the dead snake, 185 by identification of the snake in a photograph]. 273 (76.5%) of the 357 admissions were due to hump nosed viper bite. AVS was given to 184 (39.5%) patients, including 99 (36.3%) with hump nosed viper bite. A sensitivity test of AVS was used in all 184 patients. Premedication with hydrocortisone and/or antihistamines before AVS infusion was given to 89 patients. Acute adverse reactions to AVS occurred in 102 (55.4%) patients given AVS. There was no significant difference in the rate of reactions whether premedication was given or not. CONCLUSION: Precise identification of the offending snake was not possible in many instances. Practices that are of no benefit in the treatment of snake bite are still widely used. Acute adverse reactions to AVS are common, and neither hydrocortisone nor antihistamines seem to be of benefit as prophylaxis. Evidence based management guidelines, especially with regard to AVS therapy, are urgently required. PMID- 11051704 TI - Female karyotype (XX) in patients with ambiguous genitalia does not guarantee the absence of intra-abdominal testes. AB - We report three patients with ambiguous genitalia and 46 XX karyotype who had intra-abdominal testes. The mechanism for the development of testes in such patients, the risk of malignancy and value of histological assessment of intra abdominal testes are discussed. PMID- 11051705 TI - A trial of deferiprone in transfusion-dependent iron overloaded children. AB - OBJECTIVE: To determine the efficacy and safety of deferiprone. DESIGN: Prospective study. SETTING: 5 paediatric medical units at the Lady Ridgeway Hospital for Children (LRHC), Colombo. PATIENTS: Transfusion-dependent iron overloaded children in the age group 2 to 15 years. INTERVENTION: Patients were given a total daily dose of 75 mg/kg of deferiprone orally in divided doses. MEASUREMENTS: Efficacy of deferiprone therapy was assessed by 4-monthly serum ferritin assays using the ELISA technique. Safety of deferiprone therapy was assessed by 4-weekly white cell counts, platelet counts and serum transaminase levels. The Z-test was used to assess the significance of the difference between the mean initial serum ferritin level and the mean subsequent serum ferritin level. RESULTS: 54 patients received deferiprone therapy for a mean duration of 9 +/- 3 months. Initial serum ferritin levels ranged from 1500 to 10,700 ng/ml with a mean of 5743. Subsequent serum ferritin levels, obtained in 48 patients ranged from 740 to 7300 ng/ml with a mean of 3558 (p < 0.001). In 47 of the 48 patients subsequent serum ferritin levels were lower than initial levels. One child developed severe neutropaenia, which reverted to normal on discontinuation of treatment. 11 children developed arthropathy, which responded to ibuprofen therapy combined in some cases with a reduction of the dose of deferiprone to 50 mg/kg/day. Serum transaminase levels were raised in 5 patients but reverted to pretreatment values or lower despite continuation of deferiprone therapy. CONCLUSIONS: Deferiprone is a safe and effective oral iron-chelating agent which can be used, under strict supervision, in transfusion-dependent iron overloaded children. PMID- 11051706 TI - Urinary tract infection and antibiotic susceptibility in malnourished children. PMID- 11051707 TI - Juvenile pernicious anaemia in a Sri Lankan child. PMID- 11051708 TI - Synovial sarcoma is an unusual site. PMID- 11051709 TI - Complex obstetric fistulae--two case reports. AB - Two cases of complex obstetric fistulae are reported. Poor assessment and mismanagement of labour were revealed. The resulting fistulae were vesico-urethro rectovaginal and vesico-cervico-vaginal. Plastic repair using a modified Martius graft was used in case 1. Considering the young age of the patient, transplantation of the ureters into the rectum or colon was deemed undesirable. The patient's endurance over a period of ten years, with seventeen attempts at repair ultimately was rewarded by achieving both vesical and rectal continence. Repair using the transvaginal route was successful at the first attempt in case 2. PMID- 11051710 TI - Say 'no' to shackling prisoners in hospital. PMID- 11051711 TI - The drug regulatory authority. PMID- 11051712 TI - Cost-effective drugs? It depends on what door is chosen. PMID- 11051713 TI - Blood pressure measurement by final year medical students. PMID- 11051714 TI - Intra-amniotic injection or intra-uterine injection of methylene blue? PMID- 11051715 TI - Intra-amniotic injection or intrauterine injection of methylene blue? PMID- 11051716 TI - [Population genetics of Chinese surnames. II. Inheritance stability of surnames and regional consanguinity of population]. AB - This paper analyzes and compares the distributional curves and isonymy of surnames and the consanguinity of regional population in the Song and Ming dynasties and the 2 present. The distribution of surnames in the three periods reveals two significant phenomena: (1) The historical inheritance of Chinese surnames is continuous and stable. This explains why the consanguineous culture relics represented by surnames and the evolution of life substances especially Y chromosome has basically the same pattern. (2) Two types of surnames, common and rare, can be identified in China. The 100 common surnames, less than 5% of the total number of Chinese surnames, are connected with more than 85% of the population, while the rare surnames, more than 95% of the total number of surnames, are related to only about 15% of the population. The distribution of common surnames acts as the major factor reflecting the genetic composition in different regions, and it determines the historical population migration and the degree of consanguinity between regional populations. The rare surnames are of regional characteristic and relative isolation. As a result, it is possible that the study of Chinese surnames and of the distribution pattern of population with the same surname serves as an important approaches to Chinese paternal genetics and Y chromosome evolution. This may provide valuable clue for the study of population highly subject to genetic diseases. PMID- 11051717 TI - [A study of transgenic IFV cattle integrated with human serum albumin gene]. AB - The mammary gland expression vector (pcDNA 3.1-GCALBm) containing the full-length sequence of human serum albumin (hALB) cDNA and intron 1 as well as the goat beta casien gene promoter and 5' up-stream regulatory sequence was constructed. The vector was micro-injected into bovine IVF eggs. The embryos were in vitro cultured to the late stage of morulae, and then few embryo cells were aspirated for the implantation detection of target gene integration and SRY DNA sequence using nested-PCR. Afterwards, ten integrated embryos were selected to transfer into eight recipients and three were pregnant. The pregnant rate was 37.5%(3/8). However, two were miscarried in mid-trimester but one was pregnant at term to deliver a male transgenic cattle integrated with hALB mini-gene. The transgenic efficiency was 12.5% (1/8). PMID- 11051718 TI - [Prediction and evaluation of heterosis of beef cattle and their application]. AB - The genetic structure and genetic variation of eight beef cattle cross parents populations were analyzed by six microsatellite loci, and heterosis of beef cattle was predicted. On the basis of microsatellite analysis, the effect of 18 cross combinations was estimated by the method of individual animal model BLUP. A new method of molecular quantitative genetics that select best of all cross combination was submitted. The results showed that the combinations with Hereford Limousine and Charolais as paternal parent are better than others in Fengning and Longhua regions; the combinations with Limousine Angus and Hereford as paternal parent are better than others in Zanhuang regions; the combinations with Hereford Limousine and Piemontese as paternal parent are better than others in Funing regions. Effect of three breeds cross is better than two breeds. PMID- 11051719 TI - [The effect of marker density on QTL mapping in a grand-daughter design]. AB - Using Monte Carlo method, the effects of marker density on the accuracy of QTL mapping, measured with mean squared error (MSE) of QTL position estimates, were investigated for a grand-daughter design under different population structures, heritabilities, sizes of QTL effect, and QTL locations. The optimal marker densities for QTL mapping for applying to marker assisted selection, (MAS) were analyzed from an economic point of view. The results show that, in general, with the increase of marker density MSEs decrease much more rapidly when all other factors are at a high level than at a low level. When the sample size reaches a certain level (e.g. 40 families with 100 sons per family), or when the QTL effect is large (e.g. the QTL explains 50% of the genetic variance), the decrease of MSE will be only slightly affected by other factors. For the best benefits of MAS, the optimal marker distances under different combinations of various factors were 5cM (small sample and large QTL, moderate sample, heritability and QTL, or large sample and small QTL), 10cM (moderate sample and high heritability or large QTL, or large sample and moderate heritability and QTL), and 20cM (large sample and high heritability or large QTL). PMID- 11051720 TI - [QTLs and epistasis underlying rice (Oryza sativa L.) panicle length in different genetic background and environments]. AB - A double haploid (DH) population and a recombinant inbred (RI) population derived from a cross between a japonica male parent Azucena and indica female parents, IR64 and IR1552 respectively, were used in both field and pot experiments for detecting QTLs and epistasis for rice panicle length in different genetic background and different environments. Panicle length (PL) was measured at maturity. QTLs for PL were detected using single marker analysis and interval mapping. Epistasis effects on the trait were also analyzed. Nine QTLs were detected in DH population, including 5 QTLs detected from field experiment and 4 from pot experiment, among them 3 QTLs mapped on chromosomes 1 and 4 were identified in both field and pot experiments. No significant epistasis effect was detected for PL. Four QTLs were detected in RI population, among them 2 from field condition and 2 from pot experiment. 6 pairs of epistasis loci were detected in RI population. One QTL mapped on chromosome 4 and two pairs of epistatic loci were detected in both field and pot experiments. One QTL on chromosome four was identified in both populations, and one marker locus RG323 on chromosome one was involved in additive effect in DH population, but epistasis effects in RI population. PMID- 11051721 TI - [Tissue culture induced translocation conferring powdery mildew resistance between wheat and Dasypyrum villosum and its marker-assisted selection]. AB - Glutamate oxaloacetate transaminase (GOT) electrophoretic analyses were performed in 175 regenerants arising from immature embryos of crosses between wheat (Triticum aestivum L.) and 6D/6V substitution stocks. The GOT-V2 coding specific enzyme band was absent in two regenerants, designated 98R149 and 98R159 respectively, originated from cross of Yi 4095 and 6D/6V substitution stock c.v. RW15. Pm21 gene linked SCARs (Sequence Characterized Amplified Regions) analysis indicated that 6VS chromosome arms existed in 98R149 and 98R159. Fluorescence in situ hybridization with total genomic DNA extracted from Dasypyrum villosum (L.) as a probe confirmed the occurrence of translocation between 6V chromosome and a wheat one in the two regenerants mentioned above. 98R149 and 98R159 were immune to powdery mildew (Erysiphe graminisDC. f. sp. tritici) inoculation with mix races collected from Hebei Province. The results of the present paper added another feasible example of useful translocations via tissue culture. PMID- 11051722 TI - [Molecular cytogenetic characterization of a new wheat line YW443 with resistance to barley yellow dwarf virus]. AB - A new wheat line YW443 with BYDV resistance and good traits was developed from the combination of PP9-1/Shan7859?Fengkang 8. The wheat line YW443 was identified by BYDV resistance analysis, genomic in situ hybridization (GISH), RFLP and RAPD analysis. The results indicated as follows: (1) YW443 was resistant to GPV and GAV strains of BYDV; (2) YW443 is a homozygous wheat-Thinopyrum intermedium translocation line with a pair of BYDV resistance genes; (3) The 7StL segment of Th. intermedium carring BYDV resistance gene was transferred onto the distal end of the wheat chromosome 7D long arm, the line YW443 was 7DS.7DL-7StL translocation; (4) A RAPD marker OPR19(-900) could detect the DNA of Th. intermedium 7StL in L1 and L1 derivatives including translocation lines YW443, YW642 etc. The markers may be used as a selecting marker for the BYDV resistance breeding program. PMID- 11051723 TI - [Molecular proof of wheat transformed by total DNA of Leymus racemosus]. AB - Four repetitive DNA sequences (pHv7161, pHv7179, pHv7191 and pHv7293) cloned from barley (Hordeum vulgare) genome were used for a molecular hybridization in the genome of Leymus racemosus (donor), Spring wheat 761 (receptor) and the wheat transformed by total DNA of Leymus racemosus (donor) through pollen tube pathway. Our results indicate that some bands are common in Leymus racemosus and the transformed wheat but absent in Spring wheat 761. The HindIII fragments which are common in Leymus racemosus and the transformed wheat have been cloned. The clone of Leymus racemosus was nominated as pLR980. pLR980 contains the homologous sequences to the four repetitive sequences of barley. The pLR980 cloned from Leymus racemosus was used as probe to study Leymus racemosus, Spring wheat 761 and the transformed wheat. This study demonstrate that pLR980 is homologous in both genome of Leymus racemosus and wheat. Common bands absent in the Spring wheat 761 were also showed in Leymus racemosus and the transformed wheat. pLR980 was proved to be associated with the transform of wheat by total DNA of Leymus racemosus through pollen tube pathway. This provides a direct witness to the exogenous DNA introduction. PMID- 11051724 TI - [Soybean germplasm diversity and genetic variance detected by microsatellite markers]. AB - Microsatellite or Simple Sequence Repeats (SSR) marker is a new type of molecular marker developed recently, which can be used in genotype identification, pedigree analysis, and estimation of genetic distance. Using 5 pairs of SSR primer, 21 polymorphic fragments were obtained in 15 soybean germplasm. The number of alleles at each SSR locus is from 3 to 6, and the range of gene diversity is 0.439-0.668. In addition, analysis of genetic distance was also performed in this study. Pedigree analysis showed that mutation occurred for individuals in the F8 population of RIL after multigeneration meiosis, which was caused because of the changes in the number of repeat unit, to a limited extent. PMID- 11051725 TI - [Identification of mixed major genes and polygenes inheritance model of quantitative traits by using DH or RIL population]. AB - The accuracy of the mixed inheritance analysis of quantitative traits with larger experimental error could be improved while using DH or RIL population. The segregation analysis method of identifying mixed major genes and polygenes inheritance model, including linkage inheritance model, of quantitative traits by using DH or RIL population was developed in this paper. The method may be applied to identify the mixed major gene and polygenes inheritance model of quantitative traits, estimate genetic effects and variances of major genes and polygenes, and the recombination value while there is linkage between two major genes. Finally, an example was used to illuminate the above procedure. PMID- 11051726 TI - [Isolation and characterization of PAOX2 mutant in Pichia pastoris]. AB - Spontaneous Mut+ mutants of P. pastoris AOX1-defective expression strain have been isolated, they were identified as phenotypically utilized methanol to grow as wild type. The results obtained from measuring growth curve when cultivated in medium in which methanol as a sole carbon source and detecting HSA protein on SDS PAGE confirmed that the mutants have increased ability to utilize methanol and express foreign HSA gene product. The promoter region of AOX2 gene from the mutants has been cloned by PCR amplification, and the DNA fragment is 1022bp in size. Sequencing analysis showed that there are two point mutations at positions of -529 and -255 from the translation initiation codon respectively. The mutations improved AOX-1 defective function and facilitate the foreign gene for higher expression. PMID- 11051727 TI - [Construction of B. thuringiensis shuttle vector and expression of the cry1C gene]. AB - We have constructed the E. coli-Bt shuttle vector pHV-1 by cloning the replicon (approximately 1.6 kb) of Bt ken-Ag and the aphI gene of pUC4K into pUC19. The rate of plasmid maintenance is more than 80% after 100 generations in E. coli, whereas 80% after 40 generations in Bti 4Q8. We have also constructed pHV-cry1C through cloning the alpha-amylase promoter from B. licheniformis and the cry1C gene from Bt 9510 into pHV-1 and introduced it into Bti 4Q8 by means of electroporation. Under the microscope, we can see that there is no crystal in Bti 4Q8, however, there are many rhomboid crystals in Bti 4Q8 (pHV-cry1C), which are smaller than those of Bt 9510. The bioassay result of Bti 4Q8 (pHV-cry1C) demonstrates that the expressed crystal protein is insecticidally active against Spodoptera exigue. PMID- 11051728 TI - [Translation initiation function of the regulation element in the operon of cholera toxin A]. AB - To demonstrate that there existed translation coupling between cholera toxin A subunit gene and B subunit gene, and give the answer why the expression level of B gene is five times more than that of A gene, alpha report system for the investigation of translation coupling was constructed by using lacZ gene as reporter. Frame-shift mutation was introduced near the C terminal of ctxA gene, and the ribosome would read through its normal stop codon. The report plasmid was constructed and it was found that the expression level of lacZ gene decreased five times after the frame-shift mutation. The translation of cholera toxin B subunit gene was translational coupled with A subunit gene, and was responsible for the differential expression level of the two genes. PMID- 11051729 TI - [The development of the concept of "glomerulonephritis"]. PMID- 11051730 TI - [Pathogenesis, diagnosis and treatment of subclinical hypothyroidism]. PMID- 11051731 TI - [The Helsinki Accords of the World Medical Association: guidelines for physicians concerning biomedical experiments on humans]. PMID- 11051732 TI - [Hypertrophic cardiomyopathy and viral, idiopathic myopericarditis]. AB - Among 64 patients followed up for 3 to 24 years after acute viral or idiopathic myopericarditis 10 patients were found to have hypertrophic cardiomyopathy. At echo-CG they had asymmetric hypertrophy of the septum. Manifestations of acute myopericarditis, results of laboratory and functional investigations are presented. Hypertrophic cardiomyopathy runs asymptomatically or with few symptoms. The relations between cardiomyopathy and viral infection are discussed. PMID- 11051733 TI - [Differential diagnosis of hemotamponade in myocardial infarction and aortic dissecting aneurysm]. AB - Clinical picture was analysed for 29 patients with myocardial infarction and 12 patients with dissecting aortic aneurysm who had died of cardiac hemotamponade. Similar symptoms of these conditions did not allow to establish pathognomonic clinical criteria. To prevent diagnostic mistakes, it is recommended to use early device examinations. PMID- 11051735 TI - [Valsalva's test in patients with bronchial asthma]. AB - To examine feasibility of usage of phase IV hemodynamic alterations in the course of Valsalva's test in patients with bronchial asthma for detection of latent insufficiency of the right heart and pulmonary hypertension, 24 patients with mild bronchial asthma and 8 healthy individuals were subjected to Doppler echocardiography. The results indicate that this method is simple, well tolerated, can assess functional reserves of the lesser circulation. It can be used both for primary diagnosis and follow-up. PMID- 11051734 TI - [Immune disorders in periodic disease]. AB - The examination of 404 patients with periodic disease (301 with uncomplicated form and 104 with amyloidosis complication) has detected decreased functional activity of T-lymphocytes and their suppressor subpopulation, their subnormal quantity. There was a rise in T-helper/T-suppressor index, level of B- and 0 lymphocytes, sensitivity to the renal antigen. Colchicin therapy stopped the attacks and stimulated T-suppressor activity. Combined treatment with tactivin is proposed. PMID- 11051736 TI - [Osteoporotic fracture of the vertebral body in patients with steroid dependent bronchial asthma]. AB - The aim of the study was an analysis of the frequency of oral steroids prescription to patients with bronchial asthma treated in hospital and of vertebral fractures and deformations as the result of GCS-induced osteoporosis in patients with bronchial asthma taking prednisolone tablets for at least 6 months. The incidence of steroid-dependent bronchial asthma (SDBA) was 12.9%. Roentgenomorphometric analysis of the thoracic and lumbar spine performed in 52 patients with SDBA revealed vertebral fractures in 11(21.1%) patients, deformations of the first and second degree in 25(48.0%) patients. Depressed fractures of the vertebra were the most numerous--14.6%. Males and females had vertebral fractures with similar frequency (21.4 and 20%, respectively). A correlation was found between the number of the affected vertebra and prednisolone treatment duration, total dose. PMID- 11051737 TI - [The effectiveness of hyperbaric oxygenation in chronic obstructive bronchitis made in outpatient clinic]. AB - General clinical examination, external respiratory function investigation, antrovent pharmacological tests were performed in 68 patients aged 28-52 years with chronic obstructive bronchitis (COB) admitted to the rehabilitation center after hospital treatment. Hyperbaric oxygenation improved general condition of the patients, normalized excessive free-radical lipid oxidation and antioxidant defense. It also improves the results of combined rehabilitation treatment, bronchial permeability, sensitivity of beta-2 bronchial receptors to sympathomimetics. This allows to reduce doses or discontinue sympathomimetics. PMID- 11051738 TI - [Some pathogenetic mechanisms of bacterial bronchial asthma in paecilomycosis]. AB - 2-week to 18 year follow-up data are analysed for 190 patients aged 3 to 70 years (91 females and 99 males) with bronchial asthma in Paecilomyses infection. Paecilomyces fungi play the role of a specific agent responsible for development of bacterial asthma. The bronchial spasm was provoked, on the one side, by hypersensitivity with participation of IgE defending the organism from the fulgi persisting in the blood and pulmonary tissue, on the other side, by biologically active substances produced by fungal cells, exogenic phospholipase A2, in particular. Clinical characteristics, course and outcome of bacterial asthma in Paecilomyces infection depend on the resistance of immune system to hematogenic fungal infection and immune reactions running in elimination of the agent. PMID- 11051739 TI - [Diagnostic implications of extracellular matrix proteins in chronic hepatitis and hepatic cirrhosis]. AB - Enzyme immunoassay measured concentrations of basic extracellular matrix proteins, collagen type 3 (C-3) and fibronectin, in blood plasm of 119 patients with chronic hepatic diseases. 30, 16, 18, 6 and 49 of them had chronic hepatitis of minimal activity (MiACH), that of moderate activity (MACH), intermediate activity (CHIA), high activity (HACH) and hepatic cirrhosis (HC), respectively. The highest C-3 level occurred in C-stage HC, the lowest--in MiACH. Fibronectin was the highest in HACH, minimal--in C-stage HC. C-3 and fibronectin levels correlated with severity of mesenchymal-inflammatory syndrome in CH and HC; in CHIA, HACH and in B-stage HC--with cytolysis markers. A direct relationship was found between protein-synthetizing function of hepatocytes and fibronectin levels in CHIA, HACH and HC while it was inverse in relation to C-3 amount in HC. Thus, tests for plasm C-3 and finronectin expand potentialities of laboratory diagnosis of the process activity in CH and HC, allow prediction of probability of CH transformation into HC. PMID- 11051740 TI - [Upgrading efficiency and safety of combined antihelicobacterial treatment of ulcer patients using modern clinicopharmacological approaches]. AB - Therapeutic monitoring of ranitidine and omeprasol using automatic analyser REMEDi HS Drug Profiling System (Bio-Rad, USA) was performed in 120 patients with morphologically verified ulcer associated with Helicobacter pylori. The addition of antibacterial drugs elevated concentration of blood ranitidine, omeprasol being stable. The highest therapeutic effectiveness was achieved with combination of ranitidine plus metronidasol and jozamycin as well as omeprasol plus metronidasol and claritromycin. Ulcer patients with concomitant hepatobiliary diseases significantly more frequently developed side effects of antihelicobacterial therapy and the rise of ranitide and omeprasol concentrations in blood can serve a prognostic criterium of their appearance. As the highest tolerance was observed in the treatment with combination ranitidine + metronidasol + oletetrin, this regimen is recommended as the safest antihelicobacterial therapy. PMID- 11051741 TI - [Combined use of zolpidem and enalapril in elderly patients with essential arterial hypertension and sleep problems]. AB - Changes in 24-h profiles of arterial pressure following treatment with zolpidem were studied in 12 patients with essential arterial hypertension and chronic sleep problems on enalapril. One week treatment with zolpidem improved quality of sleep, increased a circadian index of systolic arterial pressure, 3 non-dipper patients recovered circadian rhythm of arterial pressure. The antihypertensive effect at night was higher when enalapril was used in combination with zolpidem than in monotherapy with enalapril. PMID- 11051742 TI - [Comparative antianginal efficiency of prolonged nitrites in patients treated surgically for ischemic heart disease]. AB - To compare clinical and cost effects of prolonged nitrates (sustac-forte), isosorbide dinitrate (nitrosorbide) and isosorbide-5-mononitrate (mono mac 40), a retrospective analysis was made of 112 cases of ischemic heart diseases (functional class II-III) treated surgically. During their rehabilitation in sanatorium 30-40 days after coronary artery bypass operation the patients received sustac forte (n = 37), nitrosorbide (n = 36), mono mac 40 (n = 39). By clinico-functional effect mono-mac appeared 1.5-2 times more effective than sustac forte or nitrosorbide. By cost-effect parameters, a course 30-day treatment was the cheapest when made with mono mac (3 times more cost effective than sustac-forte and 2 times more effective than nitrosorbide). PMID- 11051743 TI - [Comparative study of isosorbide dinitrates and mononitrates in patients with ischemic heart disease and stable angina pectoris caused by stenosing coronary atherosclerosis]. AB - In an open clinical trial 19 patients with angina pectoris (functional class II III) received in turn either non-retard tablets of isosorbide dinitrate (nitrosorbid, cardiket) in a mean dose 80 mg/day or isosorbide 5-mononitrate (mono mac) in a mean dose 51.5 mg/day. Each drug was given for a month. The effect was assessed by changes in frequency of anginal attacks and exercise tolerance. Non-retard isosorbide dinitrate and isosorbide 5-mononitrate demonstrate a good antiischemic effect, are safe and well tolerated. Isosorbide dinitrate and mononitrates do not differ significantly in reduction of the anginal attacks and by an increase in exercise tolerance but the effective dose of mono mac was 1.5-2 times less than that of nitrosorbide or cardiket, thus it is more cost-effective. PMID- 11051744 TI - [Quamatel in the treatment of acute and chronic pancreatitis]. AB - The aim of the study was to assess effects of H2-blocker of the IIId generation famotidine (quamatel) on the progression of acute and chronic pancreatitis and intensity of pain in patients with pancreatitis. 46 patients received standard therapy plus intravenous quamatel. Control patients received only standard treatment. It was found that intravenous quamatel relieves pain primarily within the first treatment week. Quamatel shortens duration of analgetics use, narcotic, in particular. It is thought valid to include quamatel in combined treatment of patients with acute and chronic pancreatitis. PMID- 11051745 TI - [Generalized echinococcosis]. PMID- 11051746 TI - [Current activity of pulmonological consultative commission]. PMID- 11051747 TI - [Approaches to improving quality of life for patients after gastrectomy and subtotal distal resection of the stomach]. PMID- 11051748 TI - [Can fungal thyroiditis be casuistic?]. PMID- 11051749 TI - Oral Health Forum 2000: Opening address by the Minister of Health, the Honourable Annette King. PMID- 11051750 TI - Keynote address: Principles of oral health services planning. AB - The opportunity exists in the Forum to put together a new vision for the future of oral health for New Zealanders and for an appropriate system to meet their future needs. If we can develop a shared view of that future, it is a simple matter of planning, discussing, and compromising, but moving toward that vision. If we do not have a vision of where we want to be, we have no idea of how to get there. We need a vision of "oral health services in New Zealand" in the private sector, in the public sector--on the West Coast of the South Island of New Zealand, in the specialist practices of Auckland and Christchurch, in rural practices in Waimate and Waihi, on marae from Te Aupouri to Kati Mamoe, in the Executives of the New Zealand Dental Association and the Dental Council of New Zealand, in Government, in district boards to community halls in Mangere. Your vision of the future for oral health and oral health services in New Zealand should be known, talked about, and shared as a national value. If you can achieve nothing more than a shared vision, you have achieved a lot. PMID- 11051751 TI - Proposal 1: Do we have the right work force and delivery system? Do we have the right work force mix and delivery system? PMID- 11051752 TI - Oranga niho: Maori oral health services. PMID- 11051753 TI - Oral health services: externalities and implications for practice. PMID- 11051754 TI - Proposal 2: How do we enhance the consumer-provider partnership? The consumer provider partnership. PMID- 11051755 TI - Proposal 3: Is a coordinated and effective approach to work force planning, education, and research possible? Part I: The perspective of the school dental therapist. PMID- 11051756 TI - Proposal 3: Is a coordinated and effective approach to work force planning, education, and research possible? Part II: The interdependence of research, education and the oral health work force. AB - Undoubtedly, to achieve the goals set for oral health in New Zealand, representative leadership is essential for effective coordination of work force planning, education, and research. It is to be hoped that, from discussion and debate, this Forum will be able to make suggestions or recommendations for a structure that can be developed to allow the appropriate representation and leadership to drive research and work force planning for the provision of oral health care in New Zealand. PMID- 11051757 TI - Proposal 4: What oral health data do we need in order to plan, and how do we collect it? Part I: The epidemiologist's perspective. PMID- 11051758 TI - Proposal 4: What oral health data do we need in order to plan, and how do we collect it? Part II: The private practice perspective. PMID- 11051759 TI - School dental service. PMID- 11051760 TI - Structure, mechanism, and evolution of the mRNA capping apparatus. PMID- 11051761 TI - Folding of a nascent peptide on the ribosome. AB - Even though very significant progress has been made recently in elucidating the structure of the bacterial ribosome and topological assignments of its functional parts, the molecular mechanism of how a peptide is formed and how the nascent peptides is folded on the ribosomes remains uncertain. Here, the current progress and remaining problems are considered from the standpoint of the authors. Topics considered include formation of peptide bonds and models that represent this process, the vicinity of RNA to the nascent peptide, the cotranslational folding hypothesis, evidence that some but not all nascent peptides pass through a region within the 50S ribosomal subunit, presumably the tunnel, in which they are folded and sheltered, pause-site peptides, and the involvement of chaperones in folding of nascent proteins on ribosomes. The chaperone-like activity of the large ribosomal subunit in renaturation of denatured proteins is reviewed. It is concluded that cotranslational folding of some but not all nascent peptides occurs in the large ribosomal subunit. It is suggested that this folding is facilitated by changes in the conformation of the ribosome that are related to the reaction cycle of peptide elongation. PMID- 11051762 TI - Exoribonucleases and their multiple roles in RNA metabolism. AB - In recent years there has been a dramatic shift in our thinking about ribonucleases (RNases). Although they were once considered to be nonspecific, degradative enzymes, it is now clear that RNases play a central role in every aspect of cellular RNA metabolism, including decay of mRNA, conversion of RNA precursors to their mature forms, and end-turnover of certain RNAs. Recognition of the importance of this class of enzymes has led to an explosion of work and the establishment of significant new concepts. Thus, we now realize that RNases, both endoribonucleases and exoribonucleases, can be highly specific for particular sequences or structures. It has also become apparent that a single cell can contain a large number of distinct RNases, approaching as many as 20 members, often with overlapping specificities. Some RNases also have been found to be components of supramolecular complexes and to function in concert with other enzymes to carry out their role in RNA metabolism. This review focuses on the exoribonucleases, both prokaryotic and eukaryotic, and details their structure, catalytic properties, and physiological function. PMID- 11051763 TI - Protein traffic in bacteria: multiple routes from the ribosome to and across the membrane. AB - Bacteria use several routes to target their exported proteins to the plasma membrane. The majority are exported through pores formed by SecY and SecE. Two different molecular machineries are used to target proteins to the SecYE translocon. Translocated proteins, synthesized as precursors with cleavable signal sequences, require cytoplasmic chaperones, such as SecB, to remain competent for posttranslational transport. In concert with SecB, SecA targets the precursors to SecY and energizes their translocation by its ATPase activity. The latter function involves a partial insertion of SecA itself into the SecYE translocon, a process that is strongly assisted by a couple of membrane proteins, SecG, SecD, SecF, YajC, and the proton gradient across the membrane. Integral membrane proteins, however, are specifically recognized by a direct interaction between their noncleaved signal anchor sequences and the bacterial signal recognition particle (SRP) consisting of Ffh and 4.5S RNA. Recognition occurs during synthesis at the ribosome and leads to a cotranslational targeting to SecYE that is mediated by FtsY and the hydrolysis of GTP. No other Sec protein is required for integration unless the membrane protein also contains long translocated domains that engage the SecA machinery. Discrimination between SecA/SecB- and SRP-dependent targeting involves the specificity of SRP for hydrophobic signal anchor sequences and the exclusion of SRP from nascent chains of translocated proteins by trigger factor, a ribosome-associated chaperone. The SecYE pore accepts only unfolded proteins. In contrast, a class of redox factor containing proteins leaves the cell only as completely folded proteins. They are distinguished by a twin arginine motif of their signal sequences that by an unknown mechanism targets them to specific pores. A few membrane proteins insert spontaneously into the bacterial plasma membrane without the need for targeting factors and SecYE. Insertion depends only on hydrophobic interactions between their transmembrane segments and the lipid bilayer and on the transmembrane potential. Finally, outer membrane proteins of Gram-negative bacteria after having crossed the plasma membrane are released into the periplasm, where they undergo distinct folding events until they insert as trimers into the outer membrane. These folding processes require distinct molecular chaperones of the periplasm, such as Skp, SurA, and PpiD. PMID- 11051764 TI - The intrinsically unstable life of DNA triplet repeats associated with human hereditary disorders. AB - Expansions of specific DNA triplet repeats are the cause of an increasing number of hereditary neurological disorders in humans. In some diseases, such as Huntington's and several spinocerebellar ataxias, the repetitive DNA sequences are translated into long tracts of the same amino acid (usually glutamine), which alters interactions with cellular constituents and leads to the development of disease. For other disorders, including common genetic disorders such as myotonic dystrophy and fragile X syndrome, the DNA repeat is located in noncoding regions of transcribed sequences and disease is probably caused by altered gene expression. In studies in lower organisms, mammalian cells, and transgenic mice, high frequencies of length changes (increases and decreases) occur in long DNA triplet repeats. These observations are similar to other types of repetitive DNA sequences, which also undergo frequent length changes at genomic loci. A variety of processes acting on DNA influence the genetic stability of DNA triplet repeats, including replication, recombination, repair, and transcription. It is not yet known how these different multienzyme systems interact to produce the genetic mutation of expanded repeats. In vitro studies have identified that DNA triplet repeats can adopt several unusual DNA structures, including hairpins, triplexes, quadruplexes, slipped structures, and highly flexible and writhed helices. The formation of stable unusual structures within the cell is likely to disturb DNA metabolism and be a critical intermediate in the molecular mechanism(s) leading to genetic instabilities of DNA repeats and, hence, to disease pathogenesis. PMID- 11051765 TI - Molecular and cell biology of acid beta-glucosidase and prosaposin. PMID- 11051766 TI - Regulation and function of the cyclic nucleotide phosphodiesterase (PDE3) gene family. PMID- 11051767 TI - Understanding nuclear receptor function: from DNA to chromatin to the interphase nucleus. AB - The regulation of gene expression by steroid receptors is the fundamental mechanism by which these important bioregulatory molecules exert their action. As such, mechanisms utilized by receptors in the modulation of genetic expression have been intensively studied since the first identification of hormone-binding proteins. Although these mechanisms include both posttranscriptional (1) and posttranslational (2) components, the primary level of control involves direct modulation of the rate of transcription, and it is this process that has been the major focus of research in the field. PMID- 11051769 TI - Regulation of metallothionein gene expression. AB - The rapid and robust induction of metallothioneins (MT)-I and II by a variety of inducers that include heavy toxic metals, reactive oxygen species, and different types of stress provide a useful system to study the molecular mechanisms of this unique induction process. The specific expression of MT-III in the brain and of MT-IV in the squamous epithelium of skin and tongue offers a unique opportunity to identify and characterize the tissue-specific factors involved in their expression. Studies using transgenic mice that overexpress MTs or MT null mice have revealed the role of MT in the protection of cells against numerous tissue damaging agents such as reactive oxygen species. The primary physiological function of these proteins, however, remains an enigma. Considerable advances have been made in the identification of the cis-acting elements that are involved in the constitutive and induced expression of MT-I and MT-II. By contrast, only one key trans-activating factor, namely MTF-1, has been extensively characterized. Studies on the epigenetic silencing of MT-I and MT-II by promoter hypermethylation in some cancer cells have posed interesting questions concerning the functional relevance of MT gene silencing, the molecular mechanisms of MT suppression in these cells, particularly chromatin modifications, and the characteristics of the repressors. PMID- 11051768 TI - A unique combination of transcription factors controls differentiation of thyroid cells. AB - The thyroid follicular cell type is devoted to the synthesis of thyroid hormones. Several genes, whose protein products are essential for efficient hormone biosynthesis, are uniquely expressed in this cell type. A set of transcriptional regulators, unique to the thyroid follicular cell type, has been identified as responsible for thyroid specific gene expression; it comprises three transcription factors, named TTF-1, TTF-2, and Pax8, each of which is expressed also in cell types different from the thyroid follicular cells. However, the combination of these factors is unique to the thyroid hormone producing cells, strongly suggesting that they play an important role in differentiation of these cells. An overview of the molecular and biological features of these transcription factors is presented here. Data demonstrating that all three play also an important role in early thyroid development, at stages preceding expression of the differentiated phenotype, are also reviewed. The wide temporal expression, from the beginning of thyroid organogenesis to the adult state, is suggestive of a recycling of the thyroid-specific transcription factors, that is, the control of different sets of target genes at diverse developmental stages. The identification of molecular mechanisms leading to specific gene expression in thyroid cells renders this cell type an interesting model in which to address several aspects of cell differentiation and organogenesis. PMID- 11051770 TI - [Image of the month. Portal vein thrombosis]. PMID- 11051771 TI - [Pharma-clinics. How I treat ... pruritus by an antihistamine]. AB - Pruritus is a symptom originating from multiple etiologies. Its pathogeny involves various mediators including histamine. Antihistamines, particularly the anti-H1 type can improve a few pruritic disorders. However, they do not compare with specific etiology based therapies. Choosing an antihistamine relies on the balance between efficacy and unwanted side effects which are sometimes serious. PMID- 11051772 TI - [Clinical case of the month. Autosomal dominant polycystic kidney: factors in progression of renal insufficiency]. AB - Autosomal dominant polycystic kidney disease is the most frequent genetic renal disease in our population. The variability of the evolution to renal failure, sometimes in the same family has lead to suspect a role for environment factors. In spite of active research, no factor has been identified so far. PMID- 11051773 TI - [Early diagnosis of coarctation of the aorta in children: a challenge]. AB - Coarctation of the aorta presents with characteristic and distinctive physical findings. Because delayed detection of severe forms may be fatal and late detection of classic forms is associated with premature cardiovascular disease in adult life, early detection and treatment is important. However, in our survey a few patients in whom aortic coarctation was ultimately diagnosed had the correct diagnosis made by the referring physician. Our survey suggests that an incomplete physical examination explains the diagnostic failure. This survey reaffirms the need for palpation of pulses and proper measurement of blood pressure in all infants and children to facilitate early recognition of coarctation. PMID- 11051774 TI - [Antibiotics in pregnancy: importance of rational utilization]. AB - It is now clear that antibiotic treatment in the antenatal period significantly does prolong pregnancy during conservative management of preterm premature rupture of membranes and reduces neonatal infectious diseases as well as neonatal related morbidities. In the same way, prophylactic intrapartum antibiotherapy reduces the incidence of early-onset group B Streptococcus-induced sepsis. Nevertheless, on the other hand, antibiotics in the perinatal period are associated with an increase of neonatal sepsis by organisms resistant to maternally administered antibiotics. In addition, antibiotic treatment in this period of time is emerging as one of the possible sources of the dramatic increase in atopic disorders in infants and children owing to the interference with the normal process of intestinal microbial colonization. So, guidelines for using antibiotics in the perinatal period can be said as one of the major priority in public health. Antibiotics have therefore to be rightly choosen and must be used in a rational manner. Local microbial epidemiology, period of infection onset, clinical evaluation, all together allow the physician to use antibiotics, always in association, according to the "well-thought-out wager". In addition, the pharmacodynamic/pharmacokinetic relationship of each drug has to be known, in order to increase efficacy, decrease toxicity and reduce microbial resistance. It is especially mandatory in neonatology where the differences in drug distribution and drug elimination are of great concern, as compared to children and adults. The aim of this paper is to point out such very important aspects using antibiotics in the perinatal period. PMID- 11051775 TI - [The "king child" and nutritional variations or the "loving black-mail"]. AB - The "King Baby" is like a tyrant against his/her family with his/her refusing or requiring attitudes. This well known entity is analyzed in the actual social and familial context and compared with the similar situation in the countries with nutritional deficiencies. The impact of this relational problem on nutrition is detailed as especially weaning is concerned as well as the eventual consecutive and selective anorexia. A relational solution has been proposed: the "loving black-mailing" with early detection of "mothers or situations at risks". PMID- 11051776 TI - [Melatonin. I. Physiology of its secretion]. AB - Melatonin is a hormone mainly secreted by the pineal gland during the dark phase of the light-dark cycle. The best known function of melatonin in mammals is to transmit information concerning light-dark cycles playing the role of an active neuroendocrine transducer of environmental information. Although melatonin circadian rhythm is endogenous, based on 25 hour cycles, it is modulated by light dark cycle. During the day, the light signal is sent to the pineal gland through a special neuronal pathway and inhibits melatonin secretion. During the night, the last neuron of this pathway which is coming from the cervical ganglion superior releases nonadrenalin in the interstitium. Nonadrenalin stimulates melatonin synthesis through cAMP accumulation. Some factors other than light can also influence melatonin levels. Electromagnetic fields, age, male sex, Cushing syndrome, hypogonadotrophic hypogonadism, alcoholism seem to be associated with lower melatonin secretion. Female sex, hypergonadotrophic hypogonadism, sport and fasting seem to be linked to higher melatonin secretion. Some pathologies and drugs can modulate some steps of the neuroanatomic pathway of melatonin synthesis. Stress has no effect. The influence of weight and height is still investigated. Once released, melatonin can act on different organs through specific receptors (retina, supra-chiasmatic nucleus, hypophysis, brain, blood vessels, digestive tract, ovaries). PMID- 11051777 TI - [Cutaneous pseudolymphomas]. AB - Cutaneous pseudolymphomas represent an heterogeneous group of tumours resembling true lymphomas by some aspects. Most often, they are and remain benign. For some lesions, the term prelymphoma should be preferred. Searching for an etiology is important because the eradication of the cause can clear the lesion. PMID- 11051778 TI - [How I explore ... value of isotopic techniques in the detection of sepsis in valvular prostheses]. AB - Vascular graft infection is a rare but serious complication of vascular surgery. The diagnosis is frequently difficult. White blood cell imaging (111In or 99mTc HMPAO) is an adequate technique for detecting vascular graft infection. This is a sensitive and specific method and has some advantages in the detection of perioperative or low grade infection in comparison to conventional imaging. The usefulness of other techniques like the 18FDG-PET scan is studied, but not yet evaluated. We compare the different methods actually used by illustrating three case reports of vascular graft infection for whom the diagnosis was formally made by isotopic imaging. PMID- 11051779 TI - [Clinical study of the month. Prolonged insulin independence after transplantation of islets of Langerhans in a patient with type 1 diabetes: achievement of a dream?]. AB - A Canadian research team recently reported a series of 7 consecutive patients with type 1 brittle diabetes who could achieve sustained excellent metabolic control in the absence of insulin therapy after successful islet transplantation. This exceptional success, which contrasts with previous less favourable results, is probably due to the transplantation of a higher number of islets of good quality and to the use of a new glucocorticoid-free immunosuppressive regimen. These remarkable results which provide proof of the principle that islet transplantation can be reproducibly successful, at least for one year, may raise new hope in patients with type 1 diabetes. However, the problem of supply and demand of islets is huge and will require a careful, and probably difficult, selection of the best candidates to benefit from this new therapeutic approach. PMID- 11051780 TI - [Current HIV therapy and its clinical problems]. AB - HIV-specific protease inhibitors(PI) have been available in Japan since 1997. Since then, highly active anti-retroviral therapy(HAART) including two reverse transcriptase inhibitors combined with PI became the main strategy of HIV treatment. After introducing HAART, incidence of most opportunistic infections dramatically decreased, resulted a steep decline of AIDS death in Japan as well as in the United States. However, several unexpected problems related to HAART have been coming up. One is a lipodystrophy syndrome(LDS) which is a novel side effect caused by PI. Lipid disposition was noted associated with hyperlipidemia and/or hyperglycemia. Ischemic heart diseases will emerge in patients with LDS in future. Another one is inflammatory reactions to some opportunistic pathogens, such as Mycobacteria, Pneumocystis carinii, cryptococcus, and so on, occurred during course of immune reconstitution after HAART. This reaction is sometimes too severe to continue HAART and corticosteroid is often required to control the reaction. How to diagnose and how to manage the reaction are to be determined in future. PMID- 11051781 TI - [Mechanism to cause streptococcal toxic-shock syndrome]. AB - Group A streptococci(GAS) are responsible for a number of infectious diseases in humans. The development of a variety of antibiotic drugs decreased the number of bacterial infections dramatically. Streptococcal toxic shock-like syndrome(TSLS), which is now also called streptococcal toxic shock syndrome(STSS) has, however, become an important disease because it causes severe life-threatening clinical symptoms. Several reports have described STSS during pregnancy. Although a number of GAS virulence factors have been examined, the critical causative agents for STSS has not yet been identified. The clinical symptoms and pathological examinations of STSS suggest that STSS is caused by multiple factors. We need to study the pathogenic mechanism of STSS from both the bacterial and host side. Our study indicated that the lethal activity and anti-phagocytic activity examined in mice and SLO may be a major pathogenic factor in STSS. PMID- 11051782 TI - [New approaches to the analysis of ECG data]. AB - Recently, techniques for analyzing computerized ECG data have been developed in time domain, frequency domain and micro level analysis of signals. In body surface ECG mapping, reverse problem analysis from body surface to cardiac surface was investigated, but findings were not sufficient to be clinically useful. Heart rate variability analysis from Holter ECG tapes showed the utility for determination of autonomic nervous function by time and frequency domain analysis. Late ventricular potentials recorded by Signal Averaged ECG have been beneficial for predicting malignant ventricular tachyarrhythmias, as well as delayed potentials in atrial activity. QT dispersion from 12-lead ECG has been studied as one of the indicators of ventricular instability and there was some difficulty in detecting the end points of QT intervals such as that in patients with flat T waves. The recently developed automatic analysis of the Least square fit method for detecting terminal points of T wave might be optimal compared with the other methods. Micro volt T wave alternans by spectrum frequency analysis of T wave in consecutive heart beats is a unique technique for predicting cardiac sudden death, however the mechanism remains obscure. PMID- 11051783 TI - [Signal processing and quality of displaying waveform at digital EEG machine]. AB - With great advances in computer technology, the digital EEG machine was developed and has become widely used. In this paper, signal processing and quality of display of waveform at digital EEG machine were discussed to confirm its advantages and drawbacks. Signal processing on a digital EEG machine is summarized in 3 parts as follows: i) A/D conversion(sampling) of EEG signals in the head box, ii) Digital signal processing to obtain mathematically reconstructed EEG on the computer, iii) Displaying EEG on a monitor. For sampling, it was recommended that sampling rate was 200 or 256 Hz, and resolution in 12 bits or preferably 16 bits per sample. A high amplitude artifact caused EEG flattening called DC build up saturation. Montage reformatting had a clinical advantage. However, re-filtering using 15 Hz high cut filter influenced the EEG interpretation. The maximal resolution of digital EEG machine with a 17-inch CRT display using 1600 x 1200 dots was about 22 Hz. The performance of paper used for conventional EEG surpassed the performance of CRT display using digital EEG machine. It seemed that understanding the characteristics of digital EEG machine and realization of data processing were important for utilizing its functions effectively. PMID- 11051784 TI - [Signal analysis of the ultrasonic study and recording system]. AB - Signal processing of ultrasound has advanced from the analog system to the digital system, which provides improved images. Digital scan converter is one of the new advances and the recently developed full digital system is another. The recording system has also evolved. These techniques will contribute to the evolution of ultrasonic images. PMID- 11051785 TI - [Basic principles of magnetic resonance imaging]. AB - With rapid progress in MR imaging techniques, it has now become one of the important clinical imaging techniques. In this paper, the basic principles of NMR and MR imaging techniques are briefly described. Furthermore, the technical background of principal MR imaging techniques such as the spin-echo, gradient echo, fast spin-echo and ultra-fast MR imaging technique is reviewed. The principles of proton density-, T1- and T2-weighted contrast images are also described. Finally, the technical background of functional MR imaging is briefly reviewed and the typical fMR image obtained by activation of motor cortex with finger tapping is presented. PMID- 11051786 TI - [Gene diagnosis of Alzheimer's disease and related disorders]. PMID- 11051787 TI - [The practice of teleconferencing in clinical pathology--networking among cancer center-hospitals in various parts of Japan]. AB - In the last several years, we have been able to use a telemedicine system on a network among cancer-center-hospitals connected by light fibers(so-called Cancer Network) and attend teleconferences from each site in Japan at the same time using the same images. Every week we have many medical conferences using this network system. On the practical use of teleconferences, the ratio of clinical pathological images is very high, especially histopathological appreciation is important; that is, surgical slide conference, clinico-pathological conference, orthopedic tumor conference, image conferences about digestive organs, presentation of current topics in laboratory medicine and other issues are carried out by clinicopathological images. At present, images on teleconference are still-pictures and images in High Definition Television are clear and high capacity, and of sufficient quality for pathological diagnosis. However, the coincidence-rates of histopathological diagnosis among 15 pathologists between the tele-image method and direct microscopic method varied from 38-80%. It is necessary to try to experience the images of still-picture and also animated cartoon. In the near future, the present network may extend to cover a wide areas and attend to teleconferences in every medical facilities. By attending this network system, we are able to use clinicopathological information for clinical diagnosis, treatment, research and education. PMID- 11051788 TI - [Defense mechanism and immunity]. AB - Monocyte/macrophage (m phi) or mononuclear phagocytic system(MPS) play one of the key roles in defense mechanisms in human as well as animals. Informations as to their biological functions and significance in defense mechanisms or immune systems have increased. However, some of the details are still unknown and/or controversial. Prof. Takahashi lectured on some new findings: development of m phi precedes that of monocyte, factors regulating differentiation of m phi, differences between tissue m phi and monocyte/m phi and so on. He also stressed the complexity of the development and differentiation of monocyte/m phi. Prof Yamamoto lectured on a new factor, S19 ribosomal protein, which enhances migration of monocytes and connects natural immunity and acquired immunity. Following these lectures, all participants realized the complexity and also the mystery of defense mechanisms of our bodies. PMID- 11051789 TI - [A novel monocyte chemotactic factor that connects innate immunity and acquired immunity]. AB - A monocyte chemotactic factor was separated from rheumatoid arthritis synovium, and identified as a homo-dimer of S19 ribosomal protein. When S19 protein was treated with the plasma transglutaminase, an inter-molecular isopeptide bond was formed between Lys122 and Gln137, and the chemotactic activity appeared. The S19 protein dimer caused chemotaxis via the receptor on monocytes to C5a, the complement C5-derived chemotactic factor. This dimer antagonized the C5a receptor on neutrophils. This dimer was released from apoptotic cells, and functioned in the phagocytic clearance of these cells by recruiting circulating monocytes. After engulfment, the macrophages moved to regional lymph nodes, and presented apoptotic cell-derived antigens to T cells. T cells proliferated and activated B cells, and eventually the IgM antibody response was observed. Cooperation between the innate immune response and the acquired response would induce an effective host defense primarily against viral infection. PMID- 11051790 TI - A study on task-analysis of clinical pathologists as medical consultants in Nihon University Hospital--a Japanese perspective by comparison with current status in the USA. AB - To identify our role and the customers' satisfaction, the on-call consultation service records of the Department of Clinical Pathology, Nihon University School of Medicine, Itabashi Hospital (NUIH), were analyzed. Between 1995 and 1998, 1,789 consultation services were recorded, and approximately 40% were from physicians, and 50% were from medical technologists. During office hours, many physicians made contact with us at the office of clinical pathology, the clinical laboratory and other places in the hospital by various means. They asked us to interpret multidisciplinary laboratory data, and to provide the specific information that might affect clinical management. Medical technologists asked for clinical information of patients with extreme measured values and requested that we contact with physicians. In contrast, on weekends/holidays or after routine working hours, physicians sometimes requested non-automated laboratory tests such as peripheral blood smears/bone marrow smears or Gram stains. The major contents of our responses to medical technologists were concerned with blood banking and handling of instruments not to be operated in routine work. These results reconfirm that we are still required to have clinical competence for common laboratory procedures and to have the capability of interpretation of multidisciplinary laboratory data in the university hospital. Traditionally, most Japanese clinical pathologists have been focused their attention on bench work in research laboratories. However, the present study shows that the clinical pathologists need to bridge the real gap between laboratory technology and patient care. Our on-call service system can enhance the education of clinical pathologists, and improve not only laboratory quality assurance but also patient care. In addition, in response to a need for customer access to this service with a shortage of clinical pathologists, a more effective method would be to set up a proactive systemic approach in a more rigorous academic environment adopting advances in medical informatics. PMID- 11051791 TI - [Responses in a questionnaire by medical school students who participated in the new curriculum of the clinical learning in clinical pathology]. AB - The clinical learning taken by medical students are an important part of their medical education. To develop a new, effective curriculum for the clinical learning in Clinical Pathology, the instructors defined clear general instructional objectives and specific behavioral objectives, and discussed the learning strategies and evaluation methods. The medical students at our medical school took this new curriculum in Clinical Pathology in 1999. As an evaluation method of this new curriculum, we asked all students to fill out a questionnaire that asked their opinions about the length of each component in the Clinical Pathology rotation, the content of the rotation, etc. Over 80% of the respondents answered that the rotation in Clinical Pathology was useful. Ninety-six percent of the students felt that the experience and knowledge they gained in this Clinical Pathology rotation will be useful in the clinical learning in other departments. Based on the high percentage of favorable responses from the students, we concluded that the new curriculum, which was developed after intensive planning, was successful. In summary, the feedback from students who took the new curriculum in Clinical Pathology showed that this new course was well-accepted by the students and that it created an excellent relationship between the instructors and students. Some of the responses in the questionnaires will be used to improve the Clinical Pathology rotation in the future. PMID- 11051792 TI - [Ageing society and laboratory medicine]. AB - An interest in the ageing process has increased greatly with increasing the population of the aged. The goal of this interest is to improve the quality of life(QOL) in the aged. In this paper, the presidential address "Ageing Society and Laboratory Medicine" at the 46th annual meeting of JSCP in Kumamoto'99 was summarized on the important research for ageing in the past decades. The paper presented was age- and gene-related changes, the latent variation of serum constituents and lipids abnormality in the ageing process. Concerning to the definition of reference value of healthy populations and the subjects who had no combined ailments, the reference interval of individuals(intra-personal), followed 5 years categorized by age, sex, and social conditions, gave a narrow range of variation than did a larger mixed populations(inter-personal). The reference intervals set would be a more sensitive reference than is the customary "normal range" for values occurring in inter-personal. Concerning to the study of the relationship between laboratory test and activity of daily living(ADL), the higher serum levels for TP, Alb, Hb, Glu, TC were observed in the higher ADL. The basic research techniques were also evaluated in the paper. The serum lipoperoxides were correlated with serum lipoprotein free radicals which caused atherosclerosis. The higher frequency of cerebral- and myocardial-infarction in the aged were observed in the higher serum LDL-C and lower serum level of arachidonic acid(AA), eicosapentaenoic acid(EPA), and AA/EPA ratio were observed in AMI patients with lower HDL-C groups than the healthy aged. Although Alzheimer(AD)'s disease had a progressive memory loss and immobile dementia and was reported the decrease of acetyltransferase activity in the brain, decrease of serum level of free choline, lyso-phosphatidylcholine, phosphatidylcholine(PC) and sphingomyelin(SM)/PC ratio were observed in spite of keeping normal serum level of SM. The decreased serum levels of pseudocholin esterase and albumin, especially mercaptoalbumin were observed in the healthy aged with advancing age. The early diagnosis and prediction of prognosis for the latent ailments in the aged was stressed. As to the study of variations of serum protein levels in the healthy aged, variations of serum proteins were classified into three types, 1) mainly acute phase reactant proteins such as alpha 1AT increased with advancing age, 2) transporting proteins such an albumin decreased and 3) proteins with no significant variation these were useful proteins for the early finding of latent ailments. The higher increase of alpha 1AT/beta 2III in the healthy aged over 60 y.o. was suspected to become severe in near future. PMID- 11051793 TI - [Consideration on neuroimaging and functional diagnosis in cerebrovascular disorder]. AB - The electroencephalogram(EEG) has been one of the important clinical examination in cerebrovascular disorder(CVD). Since X-ray computed tomography(CT) was introduced in the 1970's, the numbers of EEG examination decreased in Japan. In the present, the diagnosis of acute CVD is done by neuroimaging examinations which generally are CT, magnetic resonance imaging(MRI) and at times, single photon emission computed tomography(SPECT). But EEG examination is necessary for estimate of brain function in acute and/or chronic CVD. Only EEG examination is able to confirm epileptic seizure, sleep-wake disorder and disturbance of consciousness. alpha-coma and spindle coma are peculiar disturbance of consciousness and appear in acute CVD. REM sleep behavior disorder(RBD) sometimes appear in chronic CVD. The characteristic of RBD is violent or injurious behavior during REM sleep. Polysomnography demonstrates abnormal REM sleep without loss of atonia. Both neuroimaging and functional diagnosis have been to perform to estimate in CVD same as in the other central nervous disorders. PMID- 11051794 TI - [Neuroimaging and neurofunctional diagnosis of brain tumors]. AB - Recent advances in diagnostic modalities such as computerized tomography(CT scan), magnetic resonance imaging(MRI), and electrophysiological examination including EEG, SEP, ABR, have facilitated the diagnosis of relatively small brain tumors, while refinement in microsurgical techniques with the aid of intraoperative echo-sonography, endoscopy, navigation surgery and application of intraoperative monitoring of brain function, have facilitated precise diagnosis and safe surgery for brain tumors. This paper presents the preoperative, intraoperative, and postoperative usefulness of these modalities in treating brain tumors. PMID- 11051795 TI - [Neuroimaging and functional analyses for multiple sclerosis]. AB - For neuroimaging studies of multiple sclerosis(MS) lesions, magnetic resonance imaging (MRI) is most useful, while evoked potentials(EPs) are commonly used for functional analyses of neural damage due to MS. This review summarizes the MRI and EP findings in MS. MS lesions are visualized as high signal intensity lesions on T2-weighted images, proton density(PD)-weighted images, and fluid-attenuated inversion recovery(FLAIR) images, while such lesions demonstrate a low signal on T1-weighted images. New MS lesions are usually enhanced by gadolinium-DTPA on T1 weighted images, and the enhancement generally lasts 4 to 8 weeks. In Asian patients with MS, opticospinal MS(Asian-type MS) shows a significantly smaller numbers of brain MRI lesions than conventional MS(Western-type MS), while opticospinal MS shows a significantly higher frequency of the spinal cord atrophy on MRI than conventional MS. EPs are useful for detecting lesions located in certain portions of the central nervous system. MRI is not sensitive enough to detect small lesions in the optic nerves and spinal cord, whereas EPs are sensitive for optic nerve and spinal cord lesions. Thus, combined use of MRI and EPs is required for the diagnosis and the optimal monitoring of disease activity in MS. PMID- 11051796 TI - [Recent advances on routine urinalysis]. AB - Urine qualitative/semiquantitative tests with reagent-strip and microscopic examination(sediments) are most popular and basic clinical laboratory tests. However, the accuracy and precision of those tests have not been more reliable than that of clinical chemistry or hematology, mainly due to the instability of urine specimen and subjective analysis procedure. Automated analyzer with compact size and high quality of both reagent-strip method and sediments(formed elements or particles) have been commonly used in many laboratories and guidelines on urinalysis of JCCLS(Japan), NCCLS(USA) and ECLM(EU) are publishing in succession recently. These advances are expected to lead to a solution for those problems in the urinalysis field. PMID- 11051797 TI - [Health care in view of daily medical examination]. AB - The present situation in health care From the perspective of regional society, there are many public facilities which support the health, medication and welfare for the residents, and they operate their own service without an appropriate linkage or organized method of sharing information with each other. It is important to provide health care based on a principle with a concept of health information management by life stage. As present, such information is divided among several government agencies, namely the Ministries of Health and Welfare, Education and Labor. Infant, school medical exam, and adult or geriatric annual check-ups are under the control of the respective Ministries. As a result, we lack in communication between regional facilities and sharing information. Recent advancement in medical information systems and instruments have been remarkable. Especially after the electronic medical card will be in officially used, the medical check supporting system will gradually come into wide-spread use with easy operation. To swim with the current of the times, it is important to cooperate with organizations in other fields for practical use of personal health data. We must make an effort to establish an effective method of using computer and individual information to collect significant data. PMID- 11051798 TI - [Analysis of on-call consultations with clinical pathologists--identification of customer's satisfaction]. AB - One aspect whereby effectiveness of clinical pathologists can be measured is customer service and satisfaction. Clinical pathologist should identify their customers, their processes and procedures to meet these needs to the customer's satisfaction. To identify customer's satisfaction, the records of on-call consultations with clinical pathologists were analyzed. Between January 1996 and December 1998, 1327 consultations were recorded, 40% of which were consultations from physicians, 50% from medical technologists. Physicians requested interpretation of laboratory data obtained, and clinical knowledge mainly concerning the microbiology and hematology during office hours. On holidays, physicians needed help performing emergency tests such as Gram stain and Wright Giemsa stain. During office hours, medical technologists requested clinical information concerning patients in whom unreasonable data would be reported and the contact to the clinical side. Furthermore, technologists inquired about the methodology of laboratory tests during day duty on holidays. These results indicated that the clinical pathologist in our hospital could satisfy the customer(physicians and medical technologists), by providing 1) a wide range of clinical knowledge concerning not only the laboratory medicine but clinical medicine including therapeutics, 2) capability of performing emergency tests such as Gram stain and Wright-Giemsa stain, and 3) capability of interpreting the results obtained. Although these would not be adopted in every hospital, every clinical pathologist should examine his role in the hospital. PMID- 11051799 TI - [Good laboratory management and clinical laboratory physician]. AB - Medical expenses have been increasing annually, and reducing expenses while maintaining effective medical care is desirable. In the late 1990s, Japanese government introduced policies expected to improve the medical security system. In the clinical laboratory field, some revisions such as packaging of certain tests(blanket test), separation between performance and interpretation fees for laboratory test, proper use of tumor markers, and additional fees for sample management. Japanese government also wants the clinical laboratory to return accurate laboratory test result to patients and physicians. Laboratory physicians have to make a great effort to manage clinical laboratories according to the guideline for GIOs of laboratory physicians from the Japanese Society of Clinical Pathology. The laboratory physician is the key person for good laboratory management. PMID- 11051800 TI - [The proposal for young people who wish to become clinical laboratory physicians]. AB - Based on my personal experiences for past 10 years, I would like to propose my plans for young physicians concerned with clinical laboratory medicine as follows: 1. Training periods (several years after graduated from medical school): Pursue your courses in the various fields of medicine including clinical activities. 2. Periods of fellowship(approximately about 35 years-old): During this period, it is recommended to go abroad for young scientist to add to your stock of knowledges and experiences. 3. Periods of devoting yourself to the work in the clinical laboratory medicine(approximately about 40 years-old): You are recommended to be self-reliant in the routine works and academic studies in the fields of laboratory medicine. PMID- 11051801 TI - [Importance of health care work for the clinical pathologist]. AB - Health care work should be one of the important roles of the clinical pathologist. However, we do not have specific health care programs yet. I would like to propose that the Japan Society of Clinical Pathology should become involved in the region of health care programs. In the health care programs that we discuss here, target values appropriate for healthy people to maintain their health are determined. Laboratory medicine is expected to make great contributions in this respect. I hope this session will be helpful in promoting an understanding of the importance of health care programs among our members. PMID- 11051802 TI - [What to do as a laboratory physician in the front hospital]. AB - The doctor of clinical laboratory medicine in the core hospital at a local city should grasp the situation of the clinical laboratory under present medical institute and submit measures for renovation of the laboratory. Further, the doctor should take part in medical consultation based on the laboratory data. PMID- 11051803 TI - [Private practitioner & clinical laboratory physicians]. AB - The clinical(laboratory) examiner should know how to accommodate clinical necessity. Instead of the real necessity(patient's needs), the clinical examination is dependent on the physician's own clinical skill. That is clinical laboratory examinations are ordered by the doctor's own clinical experiments and data is judged and analyzed by the mass standardized levels. Each patient's own bias must be ignored. From the medical practitioner's viewpoint(who is running his own clinic), it is desirable to have personal laboratory levels to have person-to-person and tailor-made medical management. Above all, for the futuristic medical examiner, we suggest urgent development of several parameters which will easily describe personal basic laboratory levels. (We applied for the patient of "Vessel Age" as for the new personal basic laboratory level to show atherosclerotic degeneration.) PMID- 11051804 TI - [Clinical laboratory physicians and postgraduate training for laboratory technologists]. AB - The number of clinical laboratory physicians is currently about 400, and is estimated to reach, at most, 1,500 after 20 years, even if we train 50 young clinical laboratory physicians every year. This is too few to display their abilities and to prove themselves as necessary specialists in the Japanese medical world. However, the number of laboratory technologists in Japanese hospitals and clinics is currently about 50,000. If we train excellent technologists to take part in the work of clinical laboratory physicians, we will be able to prove ourselves as necessary specialists in about 20 years. Postgraduate training for laboratory technologists is necessary to increasing their abilities and increasing the number of specialists in laboratory medicine. We consider this is the most effective method of developing Japanese laboratory medicine in the 21st century. PMID- 11051805 TI - [Evaluation of vascular endothelial function]. AB - Endothelial function was evaluated in renal and forearm vessels from patients with essential hypertension. First, the increase in renal blood flow evaluated by the clearance of para-aminohippurate, serum c-GMP and urinary NOx during L arginine infusion was significantly attenuated in essential hypertension. This attenuated endothelium-dependent vasodilation was improved by angiotensin converting enzyme inhibitor, but unchanged by calcium antagonist. Second, the increase in forearm blood flow evaluated by plethysmography during acetylcholine infusion or reactive hyperemia was attenuated in essential hypertension. This attenuation was abolished by NO synthesis inhibitor. Forearm endothelial function was improved by angiotensin converting enzyme inhibitor, daily aerobic exercise and body weight reduction by low calorie diet. In conclusion, endothelium dependent vasodilation was attenuated in renal and forearm vasculature of essential hypertensives via reduction of NO synthesis. This attenuation can be improved by several treatments. PMID- 11051806 TI - [Effect of heparin cofactor II on the antithrombin III activities measured by thrombin methods or factor Xa methods--fundamental studies and clinical studies using the plasma of pregnant women]. AB - We evaluated the effects of heparin cofactor II(HC II) on the antithrombin III(AT III) activities measured by the methods of thrombin or factor Xa. Reagents A and B were using the method of thrombin and reagent C was based on the method of Xa. Purified HC II was directly measured or indirectly measured after the dilution with control plasma. Cross reaction of HC II in AT III assay were negligible in reagent C, but substantial amount of AT III activities were measured in reagent A and B. Plasma AT III activities from full-term pregnant women were significantly higher than those from non pregnant control women in reagent A, but comparable in reagent B or C. These results indicate that AT III activities measured by thrombin methods by thrombin were overestimated in pregnant women due to the cross-reactivities of HC II. It is recommended that AT III activities would be measured by the methods of factor Xa. PMID- 11051807 TI - [Measurement of reticulated platelet using whole blood: methodology and clinical application]. AB - We established simple and rapid method to measure reticulated platelets using whole blood. After whole blood was fixed with paraformaldehyde, sample was stained with thiazole orange and analyzed using flow cytometer. %RP was significantly higher in patients with idiopathic thrombocytopenic purpura(ITP) (27.7 +/- 14.3%) compared with that of normal subjects(11.9 +/- 3.1%), and elevated in 73% of ITP patients. Furthermore, %RP reflected the status of disease very well in a patient with ITP undergoing therapy. Our method can be effectively applied for differential diagnosis and assessment of the status of thrombocytopenic disorders. PMID- 11051808 TI - [Glioblastoma with a marked elevation of total protein and albumin in the cerebrospinal fluid]. AB - We describe a patient with glioblastoma who showed very high levels of total protein and albumin in the cerebrospinal fluid. The patient was a 57-year-old man who underwent subtotal removal of a glioblastoma from the left temporal lobe. Two weeks after the operation, he had quadriplegia and a lumbar puncture was performed. The concentrations of total protein and albumin in the cerebrospinal fluid were 6000 mg/dl and 3600 mg/dl, respectively, which were more than 100 times higher than the upper limits of the reference ranges and were close to their serum levels. The magnetic resonance image suggested that these abnormal values were due to total spinal cord dissemination of the tumor. The results of the cerebrospinal fluid examination showed that the albumin concentration in the fluid might increase to the serum level and the prozone phenomenon should be kept in mind when measuring its concentration in the fluid. PMID- 11051809 TI - [Estimation of sympathetic skin response using DC servo-amplifier]. AB - The aims of this paper are to make clear the characteristics of sympathetic skin response(SSR) wave. The records of SSR wave were obtained using a DC servo amplifier with the frequency response of DC to 30 Hz and drift level of 10 microV/10 min. The SSR from the palm of 10 normal control subjects(6 males and 4 females, age range 21-25 yrs) were elicited by electrical stimulation of the median nerve at upper arm. The results that the peak latency and the duration of SSR were approximately 6.32 +/- 2.5 s and 47.5 +/- 7.5 s respectively were obtained experimentally with the amplifier, though the results of AC recordings with time constant of 0.3 s were about 1.0 s and 2.0 s respectively. It can be considered that exact peak latency and duration of SSR are much longer than those data of previous studies. These findings suggest that electric stimulation should be applied at longer intervals than 60 s for rejection of the remaining potential level of SSR and the habituation. PMID- 11051810 TI - [Production of pharmaceutical proteins with mammary gland bioreactor]. AB - Mammary gland bioreactor is a useful biological system which expresses foreign genes in the mammary gland and produces functional pharmaceutical proteins in milk. This production route is appealing for it's advantages, such as the simplicity of access to the expressed protein, the high production of the mammary gland, the capabilities to perform translational modifications. As an alternative of cell culture systems, it is a new biotechnology. The article reviews some aspects on generation and characterization of mammary gland bioreactor, separation and purification of foreign protein from milk and some questions that need to be answered on the route. PMID- 11051811 TI - [Role of heat shock protein-peptide complexes on tumor and infectious diseases immunity]. AB - Many heat shock proteins, e.g. gp96, HSP90, HSP70, etc have elicited rejection and immunotherapy immunogenicity of tumor and infectious diseases. Further study indicated that hsps can chaperone the endogenous repertoire of peptides, and the antigenicity of hsp-peptide complexes lies in the peptides, not HSPs. HSPs present peptides associated with them to MHC class I molecules for recognition by CTL and memory T cells, and elicit cellular immune responses. The latest finding shows that gp96 may present antigenic peptides directly to T lymphocytes functionally as MHC. In recent years the mechanism of immunogenicity and advantages as vaccine therapy of gp96 and HSP70, the two main hsps in mammals have been studied in detail, which offers a new opportunity for immunotherapy of hepatitis B and hepatocellular carcinoma. PMID- 11051812 TI - [Imaging ellipsometry in biomolecule research]. AB - Imaging Ellipsometry is an optical surface-sensitive method for the investigation of various aspects of biomolecules adsorbing mainly at reflecting metal surfaces and silicon surface. It has advantages of high sensitivity to layer-thickness, big area of view, high sampling speed, and high lateral resolution. It can be used for studying adsorption kinetics of biomolecules and detecting complex layer of antigen-antibody when antibody (or antigen) combined with antigen (or antibody) coated on solid surfaces. Compared with other solid phase methods such as enzyme linked immunosorbent assay, immunofluorescence and radioimmunoassay, imaging ellipsometry has the advantage of not involving any labelling of reactants and it is a relatively inexpensive method and easy to handle. Compared with traditional ellipsometer, imaging ellipsometry shows an advantage of distinguishing both affinity and non-specific binding in different area on the surfaces. PMID- 11051813 TI - [Construction of a transformation-competent artificial chromosome (TAC) library of a wheat-Haynaldia villosa translocation line]. AB - Transformation-competent artificial chromosome (TAC) vector is able to clone and transfer large DNA fragments in plants and is a powerful tool for plant gene isolation and transformation. To clone important genes from wheat, a TAC genomic library was constructed from nuclear DNA of a 6VS/6AL wheat-Haynaldia villosa translocation line that harbor the gene Pm21 for resistance to powdery mildew. The library consists of 2.1 x 10(6) clones with an average DNA insert size of 35 kb, and represents in total 4.9 genome equivalents. The library was stored as clone pools in 96-well plates, and each pool contained about 1000 clones. TAC clones containing gene(s) of interest can be screened by a pooled-PCR/colony hybridization strategy. PMID- 11051814 TI - [Complete nucleotide sequences and genome structure of two Chinese tobacco mosaic virus isolates deduced from full-length infectious cDNA clones]. AB - The complete nucleotides of two Chinese tobacco mosaic virus (TMV) isolates, TMV Cv (vulgare strain) and TMV-N14 (an attenuated virus originated from a tomato strain), were determined from their respective full-length infectious cDNA clones and compared with published TMV sequences. The genome structure of TMV-Cv contained 6395 nucleotides, in which four functional open reading frames (ORF), coding for replicase (126 kD/183 kD), movement protein (MP, 30 kD) and coat protein (CP, 17.6 kD) respectively, could be recognized. TMV-N14 contained 6384 nucleotides in its genome. In contrast to TMV-Cv, five functional ORFs encoding the replicase 98.5 kD/126 kD/183 kD, MP(27 kD) and CP(17.6 kD), respectively, were detected in the TMV-N14 genome. TMV-Cv is 99% homologous to a Korean TMV isolate belonging to the vulgare strain at the nucleotide level. TMV-N14 is 99% homologous to a highly virulent Japanese isolate TMV-L (tomato strain) at the nucleotide level. In TMV-N14, one opal nulation (UGA) occurred in the replicase gene and one ochre nutation (UAA) in the MP gene. The former mutation created a potential, additional ORF within the replicase gene, the latter reduced the size of the MP to 27 kD. In addition, there were also 13 amino acid substitutions in the replicase gene of TMV-N14 when compared to that of TMV-L. Collectively, these changes may have significant implications in the attenuation of the virulence of TMV-N14. PMID- 11051815 TI - [The regulation activity of Chlorella virus gene 5' upstream sequence in Escherichia coli and eucaryotic alage]. AB - The 5' upstream regions of adenine methyltransgerase gene and major coat protein gene (PAMT, PVP54) in Chlorella virus genomes were used to contract transformation vectors in E. coli and eukaryotic algae. The regulation activities of PAMT and PVP54 comparing with PRPL and CaMV35S promoters were analyzed in different E. coli strains and Chlorella species. It is found that the luciferase activity controlled by PAMT is 50-400 times higher than that controlled by PRPL. The regulation activity of PAMT in 2 Chlorella species is obviously higher than that of CaMV35S promoter. It is the first report that the 5' upstream region of Chlorella virus gene has strong regulation activity in eucaryotic algae. The result suggests this regulation sequence will have an excellent application in the eucaryotic algae genetic engineering. PMID- 11051816 TI - [Expression and characterization of human vascular endothelial growth factor receptor Flt-1 extracellular domain in Pichia pastoris]. AB - By inserting VEGF receptor Flt-1(1-3 loop) cDNA into Pichia pastoris expression vector pPIC9K containing AOX1 promotor and the sequences of alpha secreting signal peptides, the expression plasmid pPIC9K/Flt-1(1-3) was constructed and transformed into GS115. The multi-copy insert transformants were selected and cultivated in flasks. After 4 days of 1% methanol induction, the expressed Flt 1(1-3) accumulated up to 30% of total proteins in supernatant. The expressed Flt 1(1-3) was further proved with good antigenicity and high specificity by ELISA and Western blot. They can bind to VEGF and inhibit HUVEC proliferation stimulated by VEGF. PMID- 11051817 TI - [Construction and bioassay of secondary recombinant baculovirus with two foreign gene]. AB - Plasmid pAcLEneo which bears neomycin gene derived by the baculovirus IE1 promotor was digested and the gene was harvested. A transfer vector pAc34DZ2 in which the polyhedrin envelope gene has been inactivated by insertion of expression cassette of PIE1/neo was constructed by inserting the neo cassette into the SacI site of plasmid pAc34DZ1. We have constructed the polyhedrin positive recombinant virus (I) vAcPhBtT which was able to express Bt truncated endotoxin gene. In order to improve the insecticide efficiency of recombinant virus (I), an anti-neomycin recombinant virus (II) vAcPhBtTPE- was obtained by second co-transfection into the Sf9 cells with pAc34DZ2 and recombinant virus (I) DNA. Southern blot and SDS-PAGE analysis indicated that the recombinant virus (II) was still able to express the 80 kD Bt truncated delta-endotoxin, but did not express the 34 kD polyhedrin envelope protein. The recombinant virus (II) without envelope released virion faster than recombinant virus (I) after alkaline lysis. Bioassay was carried out using Spodoera exigua. LC50 of the recombinant virus (II) was about half of wild type virus and LT50 reduced about two days. PMID- 11051818 TI - [Cloning and sequencing of the maize ribosome-inactivating protein gene]. AB - A gene encoding maize Ribosome-inactivating protein was amplified by means of PCR using mRNA of maize leaves as a template, and cloned into pUC19 vector. The amplified DNA sequence has been determined, which consists of 828 bp and encodes 275 amino acid residues. Comparison with previously reported sequence shows 98.4% homologies in nucleotide sequence and 97.4% in amino acid sequence, respectively. PMID- 11051819 TI - [cDNA cloning and sequencing of human urokinase receptor]. AB - Human urokinase receptor (uPAR), a 55 kD glycoprotein linked to the cell membrane by a glycosylphosphatidylinositol anchor, plays a central role in cell migration and tissue remodeling. The human uPAR cDNA was cloned from a highly metastatic human lung giant cell line PG by RT-PCR and then subcloned into pGEM-T vector and sequenced. The data indicate that there are three bases substitution (705, 746, 755) which subsequently leads to two amino acid mutation (249, 252) compared to that of previously reported. The cDNA sequence of uPAR was registered in GenBank with accession number AF257789. PMID- 11051820 TI - [The influences of lactose as an inducer on the expression of the recombinant proteins in Escherichia coli BL21 (DE3)]. AB - The possibility of using lactose as an inducer to substitute the common inducer IPTG in the fermentation process of the recombinant microorganism was deeply investigated. The influences of culture conditions such as lactose concentration, growth medium composition, the point of induction and the duration of the induction phase on the expression of the recombinant protein were analyzed and studied in detail. In the following experiments, lactose was then used in the high cell density culture process of E. coli BL21 (DE3)(pFu). The final cell density (OD600) was over 40. The expression level of recombinant protein was about 15% of the total cellular protein. Both the culture density and foreign protein expression level were lower than those induced by IPTG. However, because of the potential toxicity to human beings and the high cost of IPTG, the use of lactose might provide an alternative means of inducing foreign protein expression. This would be more attractive in industrial scale productions of recombinant proteins. The results confirmed that lactose could be used as an inducer in the fermentation process. PMID- 11051821 TI - [Mutations of Q20L and G247D improved the specific-activity and optimum pH of glucose isomerase]. AB - The mutants of Q20L and G247D of glucose isomerase (GI) were constructed by in vitro site-directed mutagenesis of GI gene with double-primersmethod. The recombinant plasmids pTKD-GIQ20L and pTKD-GIG247D were expressed in E. coli K38 strain. The comparison experiments of mutant enzymes with wild-type GI showed that: (1) the optimum temperature of GIQ20L was decreased by 5 degrees C. Its thermostability was only 78% half-time of the wild type. But its substrate affinity was enhanced. (2) The specific-activity of GIG247D was increased by 33%, and the optimum pH was lowered by 0.6 unit. However, the thermostability of GIG247D was decreased. We supposed, based on the above facts and 0.19 nm resolution crystal structure of SM33GI, that Gln20 locates between alpha 0-helix and alpha 1-helix, the substitution of hydrophobic side chain of Leu for hydrophilic side chain of Gln may enhance the hydrophobic interaction of the molecular surface, leading to the decrease of the stability and thermostability of GIQ20L. Gly247 which is the last amino acid of a beta-sheet from 242 to 247 residues locates in the active core of GI. After replacement, Asp247 which has strong negative electricity may change the electrostatic distribution and influence the charge transfer processes of the active core. So the specific activity of GIG247D was increased. The introduced charge could alter the pKa of dissociable groups and make the optimum pH lower. In addition, the side chain of Asp247 seems to be very crowded in the surrounding space conformation and is easy to exclude with the other side chains, therefore influences the stability of beta sheet. Furthermore, Asp247 is in the vicinity of the interface of subunits, so it could interfere with the stability of the interaction between subunits. Thus, the GIG247D decreased the thermostability of SM33GI. The higher enzyme activity and the lower optimum pH will be very useful for industrial production of GI. PMID- 11051822 TI - [Construction of expression plasmids harbouring genes encoding recombinant FN polypeptides with triple-domain and preliminary characterization of the products expressed in Escherichia coli]. AB - To investigate the important role of recombinant triple-domain FN polypeptide in tumor therapy, two expression plasmids pF94-62 and pF94-82 were constructed and used to express triple-domain polypeptides of human FN in E. coli. The expressed polypeptides were CH62 (Pro 1239-Ser 1515 of FN linked with Ala 1690-Val 2049 through Met) and CH82 (CH62 without Pro 1953-Glu 1978). CH82 polypeptide was expressed as inclusion bodies in E. coli cultured at 37 degrees C. After denaturation with 8 mol/L urea and renaturation, the polypeptides were purified by the affinity chromatograph with Heparin-agarose, and the purified product was analysed by cell adhesion assay. The expression level of CH62 in E. coli was very low(5%), but that of CH82 was very high (21%), it suggested that N terminal sequence of Cell II in FN was the key sequence which influence the expression of triple-domain polypeptide in E. coli. The purified product was capable of binding heparin and cells, and it had a better binding activity than bifunctional-domain FN polypeptides. The production of CH82 polypeptide provided a fundamental basis for further study of recombinant product with better function of anti-metastasis and immune regulation. PMID- 11051823 TI - [Breeding of arachidonic acid producting strain by ion implantation]. AB - With ion implantation, a high-yield arachidonic acid producting strain(Mortierella alpina) I49-N18 was selected, whose biomass was 30.80 g/L. The lipid component of biomass was 25.8%, in which AA content was 45.37%. The results showed that selected high-yield strain was steady on 250 L fermentor, whose AA yield was 4.0 g/L. PMID- 11051824 TI - [Studies on the production of 16 beta-methyl-11 alpha,17 alpha,21-trihydroxy-1,4 pregnadiene-3,20-dione from 16 beta-methyl-17 alpha,21-dihydroxy-1,4-pregnadiene 3,20-dione-21-acetate by Absidia]. AB - An Absidia sp. 28 strain was shown to have higher activity of 11 alpha hydroxylation using 16 beta-methyl-17 alpha,21-dihydroxy-1,4-pregnadiene-3,20 dione as a substrate. It was found that 21-acetylization of substrate could increase 11 alpha-hydroxylation rate conspicuously. The yield of the 11 alpha hydroxylation by this strain was 73% in the conversion of 0.5% concentration of 21-acetated substrate under optimum conditions. PMID- 11051825 TI - [Studies on submerged fermentation of alkaline beta-1,4-glycanases by Bacillus pumilus A-30]. AB - The effects of stirring rate, pH control and feed time of (NH4)2SO4 on beta-1,4 glycanases were investigated. The operating conditions of batch fermentation and fed-batch culture were optimized. The results showed there were a large amount of bacterial cells adsorbing on the surface of wheat bran, and stirring rate had a pronounced influence on the adsorption and the production of beta-1,4-glycanases. The uncontrolled pH could promote the induction of beta-1,4-glycanases. When (NH4)2SO4 was fed at a constant rate in the earlier age, and regulated later by determining the content of (NH4)2SO4, xylanase and CMCase activity could reach 616 IU/mL and 1.33 IU/mL respectively. PMID- 11051826 TI - [Chemical synthesis and characterization of R20A-HWTX-I, a mutant of huwentoxin-I with single residue replacement]. AB - Huwentoxin-I (HWTX-I) is a polypeptide neurotoxin purified from the venom of the spider Selenocosmia huwena. R20A-HWTX-I, a mutant of HWTX-I in which the Arg was replaced by Ala, was synthesized on solid support by using Fmoc chemistry. The synthetic mutant was oxidatively renatured in glutathione-containing buffer, and then isolated by reversed phase and specially designed ion-exchange HPLC. The chemical structure of R20A-HWTX-I was confirmed by amino acid analysis, Edman degradation and MALDI-TOF mass analysis. Physiological experiment showed that the replacement of R20 by a decreased the bioactivity of the HWTX-I by 92%, indicating that R20 is a key residue closely related to the bioactivity of the HWTX-I. PMID- 11051827 TI - [Study on protein separation using immobilized metal ion affinity chromatography]. AB - Immobilized metal ion affinity chromatography (IMAC) has shown promise of isolating desired proteins from a mixture based on their difference of affinity for chelated metal ions. With its technological superiority, such as large adsorption capacity, mild separation condition, simple ligands and wide applications, IMAC has become powerful tool for biotechnological products separation, such as proteins, amino acids and gene products. In spite of many sophisticated applications for IMAC, the theoretical analysis of Immobilized metal chromatography has remained insufficient. In this paper, the eluted efficiencies of bovine serum albumin (BSA) in a single-component system under different elution conditions are studied. The effects of several elution factors, such as pH value, ammonium concentration and anion species on protein separation are studied. Comparing the elution data of BSA in IDA-Cu and IDA-Zn columns, the different ability of affinity between metal ions and proteins is found. In addition, the elution behaviors of different proteins are investigated. This work facilitates the further research in separation of real systems. PMID- 11051828 TI - [Study on taxol release in the two-liquid-phase cultures of Taxus cuspidata]. AB - Effects of rare earth compound (ammonium sulphate), organic solvents(oleic acid and dibutylphthalate) and the integrated function of the rare earth compound and organic solvents were studied on taxol release in the Taxus cuspidata suspension cultures. And then effects of different organic solvents(paraffin, organic acid, alcohol and ester), their volumetric fraction and phase toxicity were studied on taxol release in the two-liquid-phase cultures of Taxus cuspidata. The results showed that the addition of the rare earth compound or the organic solvents could strengthen obviously taxol release, especially the organic solvents. But the addition of the rare earth compound could not strengthen further taxol release in the twoliquid-phase cultures of Taxus cuspidata. Therefore the organic solvents were very good permeabilizing reagents, which could enhance obviously secondary metabolite in the twoliquid-phase cultures of plant cells. Release percentage of taxol was increased into more than 75% from 40% of the control. PMID- 11051829 TI - [Kinetics for terephthalic acid anaerobic degradation with easily biodegradable organic material co-existence]. AB - The fact of preferential substrate utilization results in a sequence of substrate attack. As typically, the easily biodegradable substrates in TA-containing wastewater are degraded firstly through methane fermentation pathway, and just those intermediate metabolites have been proved to be inhibitors for TA biodegradation. Moreover, TA itself can inhibit the TA biodegradation, too. A kinetic model for the anaerobic digestion of wastewater containing both TA and easily biodegradable pollutants is constructed as q = qmax [formula: see text], The model parameters are estimated with non-linear regression method, the values are as follows: qmax = 1972.0 mgTA/gVSS.d; Ks = 20.2844 gTA/L; Ki,i = 2.041 gCOD/L; Ki,s = 0.0108 gTA/L. The experimental data verification for the model equation is satisfactory. According to the model analysis, a new strategy, a two step anaerobic system, dealing with this kind of wastewater is suggested. PMID- 11051830 TI - [Study on the effects of salicylic acid on taxol biosynthesis]. AB - The influence of salicylic acid on the production of taxanes in plant cell culture was studied. Experimental results showed that addition of salicylic acid at concentration of 0.1 mg/L could enhance the production of taxol to three-fold. The concentration of 10-DAB and baccatin III was also increasing while taxol concentration increases under salicylic acid elicitation. On the basis of the kinetic analysis about the simplified taxol biosynthesis pathway, a probable reason that salicylic acid improves the rate of 10-DAB producing reaction was introduced. The results above can direct its inducing mechanic research and provide the basis of multiple-elicitors synergism. The concentration of taxol arrives at 39 mg.L-1 induced by salicylic acid with silver nitrate, being 150 percent of the sum of taxol obtained under elicitation of salicylic acid and silver nitrate respectively. PMID- 11051831 TI - [The construction and expression of two humanized scFv-urokinase fusion genes]. AB - By using PCR and DNA recombination, two fusion genes of humanized mouse anti human fibrin scFv and low molecular weight single chain urokinase (Scu-PA-32K) was constructed. The difference of these two fusion genes lay in the linker between two moieties, one was (Ala)3 and another was (Gly4Ser)3. These two fusion genes were both overexpressed in E. coli with the expression level at 30%. Both expression products showed the activity of binding antigen D-Dimer and activating plasminogen after the denaturation and renaturation, but under general refolding conditions, the one with linker (Gly4Ser)3 showed better effect in the renaturation of fusion protein. PMID- 11051832 TI - [Modification of L-asparaginase with colominic acid and the new characteristics of the modified enzyme]. AB - The colominic acid was covalently coupled to L-asparaginase molecule by reductive amination. Depending on the molar ratios of colominic acid-asparaginase (30:1, 50:1 and 100:1), a modified enzyme molecule contained 4.7, 7.2 and 12 colominic acid molecule, they retained 58%, 56% and 33.2% of the initial asparaginase activity, respectively. In comparison with the native enzyme, modified enzyme had lower immunogenicity and antigenicity, longer half-life time (in vitro), more resistance ability to trypsin proteolysis, and similar Km value for L-asparagine. PMID- 11051833 TI - [Investigation on the stimulation effect of polypyrrole film on rat hepatic cells]. AB - Poypyrrole(PPy) films were prepared at 1 x 10(-3) mA/cm2 electropolymerization current density on indium-tin oxide(ITO)substrate. The PPy films were well distributed, translucent, stable and insoluble. Moreover, they can be sterilized by steam disinfection. Rat hepatic cells were cultured on these films. The results show that PPy films have good biocompatibility and they can accelerate cell growth under electrical stimulation. The cells on PPy films reach the largest cell density earlier than the cells on tissue culture polystyrene(TCPS). Furthermore, rat hepatic cells can generate on PPy films. The cells on PPy films grow faster and enter logarithmic growth phase earlier than those on TCPS. PMID- 11051834 TI - [On-line measurement of oxygen uptake rate in the cultivation of Vero cells using the dynamic method]. AB - The oxygen uptake rate(OUR) during the cultivation of Vero cells in 1.5 L CelliGen bioreactor was on-line determined using the dynamic method. The results showed that the cell growth and metabolic state during the exponential growth phase was lineally related to the OUR. This implies that the on-line measurement of OUR can be used to promptly monitor the physiological state of cultured cells and to efficiently avoid contamination because of frequent sampling in the large scale cultivation of mammalian cells. PMID- 11051835 TI - [Cultivation integrated with acetate acid filtration on Escherichia coli]. AB - A fed-batch fermentation integrated with filtration process was proceeded in a synthetic medium with the use of a hollow-fibre membrane filter. The activity of alpha A interferon reached 1.4 x 10(9) u/L, which was increased 320% over that of a control process. The integrated process was then proceeded in the synthetic medium supplemented with yeast extract during the earlier stage. The addition of yeast extract not only reduced the accumulation of acetate, but also promoted the production of alpha A interferon. The maximum activity achieved 1.9 x 10(9) u/L during the fermentation, which was increased 480% over that of a normal process. PMID- 11051836 TI - [High-level expression of human vascular endothelial growth factor in Pichia pastoris]. AB - The gene of VEGF165 was subcloned into the P. pastoris secretive expression vector pHIL-S1 and the recombinant expression plasmid pHIL-S1-VEGF165 was constructed. After transformation into yeast GS115, the positive transformants were obtained through phenotype selection and DNA Dot blotting. After induction by methanol, soluble dimer VEGF165 were expressed and secreted into the culture supernatant with its expression occupying 47% of the total protein in the supernatant. Dot blot analysis showed that the expressed human VEGF165 could bind to its receptors flt-1 and KDR. PMID- 11051837 TI - [Nucleocytoplasmic traffic and cell function]. PMID- 11051838 TI - [Nucleocytoplasmic transport of proteins]. PMID- 11051840 TI - [Nucleocytoplasmic transport in plant cells and molecular mechanism determining the localization of proteins in response to environmental changes]. PMID- 11051839 TI - [Nuclear transport of hnRNP and mRNA]. PMID- 11051841 TI - [X-ray crystal structure analysis of nuclear transport proteins]. PMID- 11051842 TI - [PTEN tumor suppressor: functions as a lipid phosphatase]. PMID- 11051843 TI - [Significance of essential completion of the Drosophila genomic DNA sequences]. PMID- 11051844 TI - [Frontiers of the human genome project: complete sequencing of the chromosomes 22 and 21]. PMID- 11051845 TI - [Technologies for tissue reconstruction]. PMID- 11051847 TI - [The historical way of the Moscow Research Institute of the Ear, Nose, and Throat]. PMID- 11051848 TI - [Clinical and morphological analysis of adenoid vegetations in children]. AB - The paper outlines the authors' clinical and morphological experience in clinically rehabilitating children with adenoid vegetations and sinusitis. Retrospective morphological and immunohistological studies have revealed that the structure of adenoids corresponds to the morphology of humoral and cellular immune responses. This enables adenoid vegetations to be regarded as an actively functioning secondary immunity organ that requires maximum sparing in young children. PMID- 11051846 TI - [Detection of in vivo protease activity using membrane-impermeant substrate]. PMID- 11051849 TI - [Analysis of the causes of vocal tract diseases in singers]. AB - The etiology of vocal apparatus diseases in opera singers is due to both specific features of their theatrical activity and conventions of opera genre in particular. A hundred and twenty seven vocalists (soloists and choristers) aged 23 to 70 years with service length of 3 to 40 years were examined. Both the status of the vocal apparatus itself and comorbidity were found to affect the occurrence of occupational diseases of the larynx. Of great importance are the quality of vocal background of a singer, his/her age, length of service, the even distribution of vocal load during a month and a season, the volume of additional work (concert and pedagogical activities), the correspondence of parts to his/her techniques and actor's capacities, and living and social conditions. The pattern of vocal apparatus diseases also depends on the type of a singer's voice and on the status of his/her nervous system. This study-based recommendations for singers and theater managers administrators reduced the incidence rates of laryngeal diseases in the followed-up professional opera singers by 15-20%. PMID- 11051850 TI - [The prevalence, causes and specific features of voice disturbances in teachers]. AB - The paper analyzes voice disturbances, their causes and specific features in teachers based on the questionnaires filled by 934 general educational school teachers. The teachers have been found to associate voice disturbances not only with changes in the voice timbre, but with different subjective feelings that make their professional activity difficult. The major factors that cause voice disturbances are the voice overloads that differ in teachers of different specialities, their inability to use the voice, psychoemotional stresses, and frequent colds, as well as a combination of several factors. The incidence of vocal apparatus diseases does not tend to decrease, which makes it necessary to implement combined medical and pedagogical prophylactic measures to prevent dysphonia. PMID- 11051851 TI - [Vocal tract disorders in women with hormonal gynecological diseases]. AB - The paper analyzes the incidence rates and specific features of vocal apparatus disorders in dyshormonal gynecological diseases in the reproductive and menopausal periods. It presents a complex procedure for diagnosis and treatment, which includes the management of the patient by a phoniatrist jointly with an endocrinological gynecologist, a psychoneurologist, and a phonopedist. Drugs for correcting sexual hormone levels, psychoemotional status, venous tone, breathing exercises and phonopedic classes by using biological feedback, and physiotherapeutical methods of treatment have been used. Indications and contraindications for the use of hormonal drugs in the treatment of dyshormonal gynecological diseases in individuals of voice-and-speech professions are considered. PMID- 11051852 TI - [The specific features of the functional status of the pituitary- adrenal and sympatho-adrenal system in children with angiofibromas of the skull base]. AB - Analyzing the results of examination of 14 boys aged 6 to 13 years who had angiofibromas of the skull base has led to the conclusion that the disease runs in the presence of the markedly activated sympathoadrenal and pituitary-adrenal systems and lipid oxidation products. While choosing a therapy, it is necessary to take into account hormonally metabolic and antioxidative imbalance and to prescribe metabolic and membrane-stabilizing drugs. PMID- 11051853 TI - [Nasal septal distortion of endogenous genesis as a manifestation of human morphofunctional asymmetry]. AB - By analyzing the data available in the literature and his own findings, the author forwards a hypothesis on the endogenous etiology of nasal septal distortions as a result of human morphofunctional asymmetry. Asymmetries are more pronounced in males and right-handed individuals than in females and left-handed ones. Due to this fact, nasal septal distortions are more common in males and with their convex part deviate to the left nasal cavity. Emphasis is laid on the necessity of using a sparing approach to removing the nasal septal cartilage during surgical interventions. PMID- 11051854 TI - [Ratio of different sound conducting ways and dynamics of Weber's experiment in health and disease]. AB - A complex of audiometric tests in ear-phones and in the free acoustic field and using hearing aids was made in normally hearing individuals and in patients with asymmetric secondary neurosensory hypoacusis in the presence of adhesive otitis media and tubal otitis. There were various frequency ranges in which the dynamics of Weber's experiment differently varied with the strength of a signal and with the type of an abnormality. The combined signal sending along the bone and air to one ear was shown to be of diagnostic value. There was a need for individually selecting characteristics of a hearing apparatus irrespective audiometric findings. PMID- 11051855 TI - [The use of cochlear autologous cartilage in surgeries for otosclerosis-induced hypoacusis]. PMID- 11051856 TI - [Diagnostic criteria and treatment strategies in traumatic injuries of the ear]. AB - The paper presents the results of examination and treatment of patients with different traumatic injuries of ear structures. The most characteristic injuries were detected in different mechanisms of traumas: mechanical (direct, indirect), altitude-induced, and thermal injuries. Major diagnostic signs of traumatic perilymphatic fistulas were revealed. The efficiency of surgical treatment for different injuries was evaluated. The surgical management of traumatic perilymphatic fistulas was comparatively assessed in different periods following injury. Evidence is provided for that it is expedient to close perilymphatic fistulas in ear injuries as early as possible. PMID- 11051857 TI - [Subjective ear noise and hyperacusis. Treatment with fluctuating currents]. AB - The prevalence of hyperacusis, and subjective ear noise in particular, tends to increase in the general population. The data available in the literature show that physiotherapeutical procedures are the most effective treatment of this abnormality. The techniques and devices using fluctuating currents were employed to treat 40 patients. A "SLUKH-OTO-1" (HEARING-OTO) device is of use now. This procedure proved to be the most effective in case of hyperacusis and subjective ear noise without hearing disorder, there was also a high percentage of positive results in chronic neurosensory hypoacusis. Treatment of subjective ear noise by this procedure is not so effective in otosclerosis and chronic otitis media. PMID- 11051858 TI - [Combined drug therapy in patients with chronic otitis media and mucositis]. AB - The data of the authors own studies using conservative therapy in patients with chronic otitis media (COM) and mucositis are presented. The efficiency of treatment of these patients is shown depending on the method of treatment (the highest efficiency being noted in the main group of patients in whom the altered middle ear mucosa was concurrently exposed to autoserum (AS) and magnetic laser therapy (MLT). The efficiency of this or that conservative treatment was found to be related to the magnitude of altered middle ear mucosal changes: the more marked the signs of mucositis are, the more difficult it is to achieve a positive result. The results of treatment were assessed visually (under an operating microscope), bacteriologically, cytologically, and crystallographically. There was a correlation between the visual assessment of obtained treatment results and crystallographic findings. The high efficiency of the proposed treatment in patients with COM and mucositis is determined by the concurrent combined AS and MLT exposure of the entire thickness of the changed mucosa and by the stimulating action of MLT on AS. This conservative therapy can pretend to be the method of choice in treating patients with COM and mucositis. PMID- 11051859 TI - [Clinical and functional studies of patients with unilateral laryngeal paralysis]. AB - Unilateral laryngeal paralysis (ULP) involves all three functions (defensive, phonating, and respiratory) of the larynx. It is necessary to make comprehensive clinical studies in patients with ULP of unknown etiology to elucidate the causes of the paralysis. The study involved X-ray tomography of the larynx, lung, and mediastinum, ultrasonography of the thyroid, X-ray study of the skull and the cervical part of the vertebral column, and barium X-ray study of the esophagus. A hundred and twenty patients with ULP underwent clinical and functional studies of the vocal apparatus, with microlaryngoscopy with palpation of the arytenocricoid joint, studies of external respiratory function and the acoustic parameters of their voice. The examination has ascertained that the clinicofunctional status of the larynx in unilateral laryngoparalysis depends on the degree of approach and the quality of closure of the vocal cords (VC) and it is determined by the position of the paralyzed VC in horizontal and vertical planes, by the degree of paralyzed VC atrophy, and by the position and mobility of the arytenoid cartilage. PMID- 11051860 TI - [The treatment of laryngeal papillomatosis with interferon inducers]. AB - Seventy three patients aged 16 to 88 years who had laryngeal papillomatosis (LP) were followed up. Microsurgical endolaryngeal removal of laryngeal papillomas was made in all the patients. The interferon inducers amixine and cycloferon as antirecurrent drugs were used in 45 patients by the regime the authors developed by taking into account the interferon status and cellular immunity of patients. The criteria for the efficiency of treatment were their improved interferon status and longer remission. The efficiency of treatment with amixine and cycloferon was 72 and 80%, respectively. Thus, the use of a sparing microsurgical intervention in combination with interferon inducers may be regarded as the method of choice in the LP treatment. PMID- 11051861 TI - [Chlamydial laryngitis: clinical picture and amixine treatment]. AB - Chlamydia trachomatis DNA was detected by PCR in 12 patients with continuously recurrent catarrhal pharyngolaryngitis. These included 9 males and 3 females whose age ranged 16 to 72 years. All the patients were found to have degrees II III alpha- and g-INF deficiencies. The interferon inducer amixine in combination with sumamed was used in the treatment of patients with chlamydial laryngitis. After treatment, there was a significant improvement in two thirds of patients. The pain syndrome was relieved, hoarseness diminished, and the laryngoscopic and microlaryngostroboscopic picture and interferon status became normal. Control studies showed no Ch. trachomatis in this group of patients. Despite positive changes in the course of the disease, the control study again detected Ch. trachomatis in 4 patients, which required repeated treatment courses. PMID- 11051862 TI - [Clinical picture, diagnosis and features of rehabilitation of the vocal tract in thyroid dysfunctions]. AB - Voice disturbances are analyzed in 105 patients with thyroid pathology. The clinical picture, diagnosis and treatment features are described in changes of the vocal apparatus due to thyroid dysfunction. A combined procedure of diagnosis and treatment rehabilitates most patients for their working vocal loads. Emphasis is laid on that patients should be managed jointly with the endocrinologist, which allows the voice to be stabilized by correcting the balance of thyroid hormones. PMID- 11051863 TI - [New method of treatment of oropharyngeal leptotrichosis]. AB - Thirty six patients with oropharyngeal leptotrichosis were examined and treated. Antistaphylococcal bacteriophage solution was used for their treatment. Tonsillar lacunae were washed with bacteriophage 2-3 times a week, the course of treatment included 6-8 washings. Thirty four patients showed clinical recovery shown as full clearance of the affected sites and elimination of subjective manifestations of leptotrichosis. PMID- 11051864 TI - [Clinical and immunological aspects of vibration therapy for sinusitis]. AB - The paper analyzes the outcomes of treatment of 326 patients with acute and chronic exudative sinusitis by using low-frequency vibratory massage- vibrotherapy that has antiinflammatory, antiedematous, and antiallergic effects, promotes normalization of body defenses and may be used as monotherapy. The therapy is shown to have a marked therapeutical effect and advantages over conventional treatments. The method is physiological, reduces the number of punctures in maxillary sinusitis, leaves the skin and mucosa intact, and decreases the use of drugs. PMID- 11051865 TI - [Antibacterial ENT treatment in outpatient setting]. PMID- 11051866 TI - [A rare case of a cyst with an osteofibrous wall in the area of the unerupted tooth]. PMID- 11051867 TI - [Some aspects of epidemiology, pathogenesis, and conservative treatment of allergic rhinitis]. PMID- 11051868 TI - [Hearing in children with specific neuropsychological features]. PMID- 11051869 TI - [Treatment of otitis in children: pilot methods with a new drug otopha]. PMID- 11051870 TI - A new century without pressure ulcers? PMID- 11051871 TI - Interface pressure measurements in leg ulcer management. AB - Interface pressure measurements are undertaken in health care as a useful tool to assist in understanding the pressures that exist between the skin surface and another surface. Pressure monitors are utilized to measure pressures under compression bandages and pressure garments and in determining pressures between patients and support surfaces such as pressure-reducing mattresses and cushions. This literature review identified that many studies using these measurements are inadequately described, and that there are many variables such as the effect of time, posture and temperature associated with such measurements that may lead to misinterpretation of results. Studies utilizing interface pressure measurements should be interpreted with caution and measurement techniques could be improved. PMID- 11051872 TI - Investigating the risk of pressure damage during childbirth. AB - The incidence of pressure ulceration is well documented within general adult hospital and community healthcare populations (Department of Health (DoH), 1992). However, within maternity units this negative outcome is largely unreported. Indeed, many trusts exclude maternity units from their regular pressure incidence and prevalence monitoring. This article will seek to raise awareness of the potential causes and areas where clinical practices could be reviewed in the light of new evidence. PMID- 11051873 TI - Treating mixed aetiology ulcers in a man undergoing drug rehabilitation. AB - Lower leg ulceration is a condition which is generally associated with the older population; however, medical conditions and social circumstances can place some clients at increased risk. This case study looks at the treatment of a patient undergoing drug rehabilitation whose leg ulceration developed as a direct result of his previous intravenous habit. It demonstrates that with correct assessment and management mixed aetiology ulceration can be successfully treated. PMID- 11051874 TI - Case study: the treatment or palliative care of pressure ulcers. AB - As nurse education increases there are reduced justifications for the existence of pressure ulcers. Patient assessment, along with rationalization of equipment and repositioning techniques, reduces the potential for pressure ulcer formation. Therefore, the future for pressure ulcer prevention will rely on nurse education and motivation. The patient featured in this case study suffered unnecessarily from pressure ulcers as, after her admission to a new nursing home where she was given the appropriate pressure-relieving and wound-dressing treatment, the ulcers were showing signs of healing. PMID- 11051875 TI - The community care of a patient with a fungating wound. AB - This case study will discuss the treatment of a malignant fungating wound and the effects that it had on a patient and his family. The article also demonstrates how interprofessional working has benefits for patients, carers and staff. PMID- 11051876 TI - Prescribing must not be at the expense of nursing. PMID- 11051877 TI - Care worker acquitted of raping vulnerable client. PMID- 11051878 TI - Case 23: the negligent practitioner. Patient who was left unaccompanied in the bath. PMID- 11051879 TI - Contraception: what has changed over the past decade? AB - The past decade has seen major advances in contraceptive technology. These have resulted in the launch of several new highly effective methods of contraception and also significant improvements in existing methods. The main purpose of this article is to review how the contraceptive field has changed over the past 10 years, explain the new methods that have become available, e.g. hormone-releasing intrauterine systems, hormone implants and female condoms, re-examine existing methods, and mention new research and how this has affected clinical practice over the past decade. When discussing contraception, clients should be given up to-date and accurate information on currently available methods: their efficacy, advantages, disadvantages and how the method works. As with any other specialty in medicine, nurses offering contraceptive advice should ensure they regularly update their knowledge and are aware of new developments and research in order to facilitate their clients in making an informed choice. New research and developments affect the choices and potentially the health of a large part of the population. PMID- 11051880 TI - Factors influencing nurses' inferences about patient pain. AB - This study examined the factors that influence nurses' perception of patient pain. Sixteen vignettes were constructed on the basis of interviews with nurses about factors affecting their perception of patient pain and previous published literature. Four variables were systematically varied within a factorial design: diagnosis (surgery vs oncology), level of patient mobility, presence or absence of physical and behavioural signs, and patient gender. Twenty-seven nurses rated each vignette for degree of inferred patient pain and degree of patient psychological distress. Mobility and positive signs of pain had statistically significant effects on nurses' judgments of inferred pain (P < 0.001), but diagnosis had no main effect. Significant interactions showed that: inferred pain in male patients was seen to be greater when immobile while mobility had no effect on the inferred pain of female patients (P < 0.003); oncology patients (compared with surgical patients) had greater inferred pain when displaying positive signs (P < 0.024); and positive signs yielded higher ratings where patients were immobile (P < 0.001). Female patients were perceived to experience higher psychological distress than men (P < 0.037) and oncology patients were perceived to experience higher distress than surgical patients (P < 0.005). Ratings of perceived pain and distress were significantly correlated but did not differ in magnitude. Offered a choice of analgesics to relieve pain for each of the patients depicted, nurses typically chose those with the lowest strength. PMID- 11051881 TI - Testicular cancer: testicular self-examination and screening. AB - Testicular cancer represents a significant threat to the health of males aged between 15 and 34 years. As there has been significant focus on early detection of cancers in order to intervene at the earliest opportunity to improve treatment outcomes, the detection of testicular cancer through secondary prevention merits careful consideration. This article discusses two health promotion approaches to dealing with the subject of optimizing men's health: the merits and disadvantages of testicular self-examination education; and screening for testicular tumours in high-risk populations. Recommendations for adapting traditional approaches are also discussed. PMID- 11051882 TI - Hepatitis B virus: cross-infection in one non-exposure prone procedure. AB - For infection control nurses (ICNs) reading about outbreaks of infection and the lessons learnt allows time for reflective practice and to change policies in one's own establishment. However, outbreak reports are not just for ICNs. One could argue that more benefit would be served if the outbreak reports were printed in non-specialist infection control journals. This article examines the hepatitis B virus and cross-infection. PMID- 11051883 TI - Confidentiality. 15: Child protection and unsuitable employees. AB - Mavis Brown learnt that Bob Downs was applying for the post of care assistant to work in the paediatric ward. She knew him from her home town where he had a reputation as a child abuser. What action should she take? PMID- 11051884 TI - To record or not to record: documentation in clinical supervision. AB - Despite the continued interest of nurses in clinical supervision there remain many unanswered questions and unresolved issues. One such key issue is that of record keeping within clinical supervision. Consequently, this article reviews the limited literature that addresses this issue. It is evident that there are three principle discrete positions regarding recording in supervision and these are summarized as (1) the superviser records minimum data to meet the needs of audit (2) the supervisee makes extensive notes for his/her learning journal, reflective diary and (3) the superviser records headings or key words to be used as an aide-memoire. The article then uses three case studies to illustrate particular concerns that the trainee supervisees have raised with regard to record keeping that centre around these three positions, and discusses the issues that arise from these concerns. As a result of this discussion, the author reasons that, when entering supervision, either as a new superviser or as a supervisee, it may well be necessary and beneficial to give mind to issues of recording. While it is unlikely that there is one 'perfect way' that will suit every practitioner, the article concludes with some general guidelines whih may help in this deliberation. PMID- 11051885 TI - Web Watch. PMID- 11051886 TI - InstantCath from Hollister: pre-lubricated self-catheterization. AB - Over the centuries a variety of substances have been used to make a hollow tube to pass through the urethra to empty a poorly functioning bladder. The ancient Greeks used dried water reeds from the riverbank. The Romans and the Egyptians experimented with gold and silver. Following the industrial revolution stainless steel was used and catheters are still manufactured today using stainless steel. PVC and plastic catheters became popular in the 1970s and various hydrophilic coatings have been added to provide self-lubrication of the catheter when it comes into contact with water. The most recent development in the clean, intermittent self-catheterization (CISC) range is the Hollister InstantCath which is a self-lubricating catheter that does not require any water. It has shown itself to be well received by patients and is a welcome addition to the choice available to healthcare professionals and their patients when instigating CISC as therapy. PMID- 11051887 TI - Karomed armchairs and cushions in the prevention of pressure ulcers. AB - The clinician providing seating for patients who are unwell, who have poor functional ability or who are at risk of pressure sores, is faced with an increasing choice of products. Making the decision as to which product and associated features to choose can be a difficult task. This article describes the importance of suitable seating provision in patients who are at risk and outlines the Karomed range of armchairs. PMID- 11051888 TI - A nursing career in the Royal Navy. PMID- 11051889 TI - PAMs are concerned about nurse consultants. PMID- 11051890 TI - Transcultural nursing and female circumcision. PMID- 11051891 TI - Is the role of circulating in an OR within the scope of practice for the RPN? AB - Role Clarity was a focal point for discussion by the St. Joseph's Health Centre (SJHC) OR Practice Council. The question: "Is the role of circulating within the scope of practice for the Registered Practical Nurse (RPN)?" was tabled. Practice Council respectfully and systematically problem solved this question, considering factors as outlined in the Decision Guide for determining the Appropriate Category of Care Provider, issued by the College of Nurses of Ontario in March 1997. The question was analysed and all involved parties reached consensus. A decision followed demonstrating that the role of the circulating nurse exceeds the scope of practice for the RPN. The RPN cannot fulfil the role in its entirety. This decision resulted in a change of practice at SJHC. PMID- 11051892 TI - Analyzing pacemaker leads: application of a form of energy. AB - In the province of Ontario, analyzing of pacemaker leads is a delegated controlled act. This article describes the certification/recertification process for analyzing of pacemaker leads at the Hamilton Health Sciences Corporation. PMID- 11051893 TI - It isn't just an alternative any more. PMID- 11051894 TI - A resource directory for all seasons. PMID- 11051895 TI - Linking care management resources and services across the continuum. PMID- 11051896 TI - Dad. PMID- 11051897 TI - Commission/URAC's new case management program standards. PMID- 11051898 TI - The Internet: a key resource for outcomes and guidelines. PMID- 11051899 TI - Increasing interaction between case management and the legal arena. PMID- 11051900 TI - Meditation. An important addition to the counselor's toolbox. PMID- 11051901 TI - Camp Heartland. Listen to my heart speak. PMID- 11051902 TI - CEU. Exploring alternative therapies. PMID- 11051903 TI - The model spinal cord injury system. PMID- 11051904 TI - A new resource for case managers working with back patients. Specialty physician networks. PMID- 11051905 TI - Certified rehabilitation professionals and direct disability costs. PMID- 11051906 TI - Practical tips to boost your chances for success. Countdown to certification. PMID- 11051907 TI - Improving recovery from chronic conditions. A discussion with Page Bailey. Interview by Patricia McCollom. PMID- 11051908 TI - The nursing shortgage. PMID- 11051909 TI - A nursing shortage like none before. Interview by Carol Lindeman. PMID- 11051910 TI - One state's view of the nursing shortage. Interview by Sonja Simpson. PMID- 11051911 TI - The changing patient is changing nursing. PMID- 11051912 TI - The ways of modern recruitment, retention. PMID- 11051913 TI - Honoring the values of practice as a retention strategy. PMID- 11051914 TI - The evolution of heart transplantation. AB - Now that heart transplantation is a common therapeutic modality, it has lost much of its earlier mystique. Numerous issues related to donor allocation, immunosuppression, quality-of-life of the recipient, patient selection, cost effectiveness, and a myriad of ethical considerations remain. Current policies, procedures, legislation, and techniques are the result of a long progression of experimental and clinical advances and public reaction. Current practices and legislation are a part of heart transplantation's evolutionary process. The process can change and advance as rapidly as it did on December 3, 1967, with the announcement of the first successful human-to-human heart transplantation. PMID- 11051915 TI - How the system functions. The roles of the United Network of Organ Sharing, the organ procurement and transplantation network, and the organ procurement organization in heart transplantation. AB - Have the limits for suitable recipient candidates for heart transplantation been exceeded? Does the current legislation and policy instituted at all levels, from DHHS to individual transplant programs, critically address the use of a dangerously limited resource? These and other questions must be the focus of future discussions regarding equitable and efficient heart transplantation in this country. The past has shown that many individuals working cooperatively within committees at federal and organizational levels have already made great strides in making organ transplantation a successful reality. Many factors influence the broadening gap between supply and demand. Each level of the system can make contributions that bring positive and creative solutions to old and new problems. Each team and committee must continue to demand representation from diverse, yet attentive, members to ensure that the specific needs of the thoracic organ recipients are properly reviewed and addressed. UNOS and its membership must continue to work together to meet the challenges of the growing acceptance of organ transplantation and the limited supply of donor organs. Continued efforts of politicians, health care professionals, and the general public must seek newer and more creative ways to manage the critical organ shortage and the ever-growing population of patients who seek heart transplantation as the only viable treatment option for their disease process. OPOs must continue their efforts to educate and promote organ donation and continue to work diligently toward increasing the pool of acceptable organ donors through improved patient management and the development of improved preservation and transport techniques. The transplant community must take the initiative to modify current legislation and to author new legislation to serve as better representors for the transplant patient population that desperately needs it. PMID- 11051916 TI - Organ procurement and the donor family. AB - The need for transplantable cadaveric organs for outweighs the supply. Improvements in the relatively new field of transplantation have dramatically improved success rates. Legislation at the state and federal levels has removed many legal roadblocks to donation. The network for the recovery and distribution of organs is well-developed and effective. OPOs, whose sole responsibility is the recovery and distribution of transplantable organs, can guide the medical staff through the process with minimal disruption of hospital routine. The recognition and referral of all potential organ donors is the responsibility of all health care professionals and helps alleviate the current shortage of transplantable organs. When simple and caring techniques are used, family consent rates for donation can approach 75%. Health care professionals remain the key to closing the imposing gap between a patient's need for transplantation and those suitable for donation. PMID- 11051917 TI - Evaluation criteria for the pretransplant patient. AB - As Achuff notes, transplant evaluation can be a lengthy and difficult process for patients and transplant personnel; however, the scarcity of donor organs mandates that heart transplant teams carefully evaluate all relevant physiologic and psychosocial data. By doing so, transplant professionals enhance their ability to select patients who are most likely to benefit from transplantation in terms of survival and quality of life, thereby making prudent use of a limited societal resource. PMID- 11051918 TI - For those who wait.... Care of the status 1 patient awaiting cardiac transplantation. AB - In conclusion, successfully bridging a Status 1 patient to the time of heart transplant requires a variety of skills. Vigilance is key in averting potential complications and addressing patient issues. Also, health care professionals will be called on to use their knowledge and expertise to deal with psychosocial and medical matters. When skillfully executed, the result of a successful transplant is an improved quality of life and a longer life. A successful transplant reflects a job well done by the professionals and is the ultimate goals for all of those who care for those who wait. PMID- 11051919 TI - The left ventricular assist device (LVAD). A bridge to heart transplantation. AB - Since its approval by the USFDA in 1998, the LVAD has been used primarily as a bridge to transplantation. It has been effective in improving the overall health and debilitated states in patients with cardiomyopathies and CHF by restoring them to a near normal hemodynamic state and improving end-organ blood flow. Recent studies indicate that the LVAD might be useful as a destination therapy, making transplantation unnecessary, providing one solution to the imbalance of heart donor supply to transplant candidate need. PMID- 11051920 TI - Heart transplantation. Long-term management related to immunosuppression, complications, and psychosocial adjustments. AB - Commendable long-term care of cardiac transplant recipients begins with excellent discharge preparation. Routine follow-up allows for prompt detection of myriad problems commonly associated with long-term survival after transplantation. Long term complications, including rejection, infection, hypertension, nephrotoxicity, osteoporosis, malignancy, diabetes, AGA, and psychosocial difficulties present unique challenges. Nursing care from the bedside, outpatient center, and home care has risen to the challenge and now must be active in the development of new strategies to maximize prevention of these many potential complications. PMID- 11051921 TI - Parental considerations for children undergoing cardiac transplantation. AB - There have been profound improvements in the survival of children following cardiac transplantation as surgical techniques, organ preservation, and immunosuppression have advanced. The sensitivity to individual parental differences and concerns throughout the beginning and long-term experience of pediatric cardiac transplantation must be accompanied by support from all the members of the transplant team. Quality of life for children, measured as functional status, at 3 years posttransplant is reported to be excellent. This quality of life is dependent not only on the medical and surgical success of the procedure, but also on an intact, supportive family. Further research needs to be done on quality of life issues after pediatric cardiac transplantation. Beyond the long-term medical, developmental, and psychological impact of the transplanted child, studies need to address the emotional, social, and financial impact of transplantation on the parents and the well siblings. PMID- 11051922 TI - "A beacon of light". Spirituality in the heart transplant patient. AB - For the cardiac transplant recipients in this study, spirituality was like a beacon of light, providing illumination and sustaining hope while enduring illness. Spirituality was nurtured by Developing Faith and by the Presence of significant others, health care providers, and the Divine to sustain hope. Spirituality throughout the transplantation process was described by concepts of faith, presence, enduring illness, and sustaining hope. The four dynamic phases of Enduring Illness can assist health care providers in understanding what spirituality means to transplant patients. This conceptual model of spirituality demonstrates the important role that spirituality plays in recovery. Clinical application of this model will allow health care providers involved in the care of cardiac transplant patients to integrate spirituality into their patients' plan of care. With future research, this model can be easily modified and built upon to meet the needs of cardiac transplant patients in all phases of transplantation. PMID- 11051923 TI - Spouse quality of life before and 1 year after heart transplantation. AB - These results indicate that overall perceived quality of life in spouses of heart transplant patients did not change significantly from the pretransplant period to 1 year posttransplantation. Conversely, specific factors influencing quality of life such as health, socioeconomic satisfaction, family satisfaction, coping styles, and the impact of the transplant experience on the spouses' life did change after transplantation. One year after heart transplantation, spouses reported less satisfaction with their health and socioeconomic status but more satisfaction with their family than they experienced before the transplant. In addition, spouses used less fatalistic, emotive, optimistic, and self-reliant coping styles after transplant than before. Lastly, spouses perceived the transplant experience more positively after the transplant than they did before the transplant. An ideal analysis would encompass data collected at more frequent periods pretransplantation and posttransplantation. The significant negative change found in the spouses' perceived health 1 year after transplant when compared with the spouses' perceived health before the transplant was based on a single-item question. A more comprehensive measure is needed to assess the health changes in the spouse that take place over time. This investigation highlights the importance of studying the impact that a catastrophic illness has on family members and the patient. As health care systems cut costs and streamline production, the needs of spouses and family members are more likely to be ignored. As a result, at some point in the future they could enter the health care arena as patients themselves. Early interventions are thus necessary to support family members of patients during the illness and throughout the recovery trajectory. PMID- 11051924 TI - Heart transplantation from an ethical perspective. AB - Ethical issues in health care have increased during the past two decades, primarily in response to rapidly evolving advanced technologies. This increase in ethical dilemmas has been most felt by nurses because they are the primary health care providers to whom patients and family members turn for support, counseling, and empathy. Heart transplantation is an area of health care that evokes numerous ethical issues. Ethical beliefs must be examined from the aspects of everyone involved: the patient, family members, and health care providers. Some aspects of the transplantation process also warrant special consideration such as transplantation, lack of a viable support system for patients, and cultural issues. Future projections include a growth in the number and types of ethical dilemmas as technology continues to evolve, populations become more diverse and older, and the health care delivery systems continue to employ a more diverse group of health care providers. This article has attempted to give nurses some insight and guidance about the ethical issues currently being encountered. The article has also included the need for future education of nurses and society. PMID- 11051925 TI - Economic trends and issues related to heart transplantation. AB - Heart transplantation continues to be a focal point of discussion pertaining to the allocation of expensive medical resources. Patients and providers currently face multiple challenges related to the financing of transplantation from pretransplant screening to long-term posttransplant care. Managed care continues to have an impact, which will hopefully lead to improved clinical outcomes and lower costs. Nurses remain instrumental as key team members and patient advocates who continue to work toward cost-effective patient management. PMID- 11051926 TI - Listening to women: focus group discussions of what women want from postnatal care. AB - Postpartum care is an essential part of the experience of childbirth and parenthood. This study explores what women want from postnatal care. Three focus groups, using a semi-structured format, were conducted. A total of 12 mothers, up to six weeks postpartum, participated in the study, which was conducted in two clinics in the Western Cape Metropole. Data was transcribed from taped sessions and analysed using Burnard's (1991) model of "thematic content analysis". Seven major categories were identified: Information, Support, Organisation of services, Attitudes of the health team, Contact with other mothers, Practical assistance and Other services. Listening to women is an essential element in the provision of flexible and responsive postnatal care that meets the felt needs of women and families. PMID- 11051927 TI - Relationship between the Whole Brain Creativity Model and Kolb's experiential learning model. AB - The aim of this article is to illustrate the relation between the cognitive styles in Kolb's experiential learning model and dominance in brain functioning. A descriptive analytical study of the literature on creativity and the development of creative thinking, explored various theories and definitions of creativity, and the nature of creative learning. Congruences between cognitive styles and the four quadrants of the Whole Brain Model were detected. This article focuses specifically on Kolb's cognitive styles in relation to the Whole Brain Model and the implications thereof for nursing education. PMID- 11051928 TI - [Standards for neonatal intensive care nursing: limited clinical nursing standards]. PMID- 11051929 TI - Student nurses' learning needs & expectations in the clinical learning units. AB - This paper describes and explores the clinical learning needs and expectations of student nurses. An exploratory, descriptive and qualitative design, which is contextual in nature, was used where a focus group interview was conducted with the final year basic students undergoing a four year comprehensive diploma course leading to registration as a professional nurse. Tecsh's (in Cresswell, 1994:155) method of data analysis was employed. Eight categories were identified as follows: communication; role modelling; up-to-date knowledge and experience; continuous supervision; assessment and evaluation; scientific process; management; professional practice and student status. A recommendation deduced from the conclusions made on the identified clinical learning needs and expectations of the students should enable teachers to address the long standing problem of how students should learn. PMID- 11051930 TI - Health education as education of the oppressed. AB - Paolo Freire's theory of critical thinking shows remarkable similarities to the principles supported by health education. In his capacity as Brazilian educationalist, Freire emphasized man's active participation in his own development. Without this active involvement, growth and development become quite impossible to attain. Freire's theory is therefore generally well suited for use by those supporters of the currently emphasised actions of community empowerment and community involvement. Health education, and more particularly successful health education, is nothing more than community empowerment. It enables each individual, within a community, to take control of his/her own life and well being. The health care professional cannot assign power and control to the individual; it can only be assumed by each individual within the community. In this regard it becomes evident that passive conveyance of the health care message, albeit from health care professional to individual recipient (in this instance to the patient), is of no use to the community, whereas active involvement of the individual displays greater success. According to Paolo Freire, health education should focus on collective knowledge, the latter being the result of the group dynamics, produced by discussion on past experiences, and the analyses there-of. The professional guides these dynamic processes, not only to identify needs regarding health education, but also to assist in making decisions on health. Ultimately each individual will then experience the responsibility for his/her own health. A discussion of the use of the theory of critical thinking, as well as the implementation there-of in practice, is provided. PMID- 11051932 TI - The Whole Brain Creativity Model: implications for nursing education and practice. AB - Recent nursing literature has repeatedly proclaimed the need for creativity in nursing. Effective nursing practice in the new millennium will require innovative and creative nurses who can adapt to change and have the courage to take risks in order to provide holistic, individualised, context-specific care. An analytical study of the literature revealed interesting information on the concept creativity and teaching for creativity. The aim of this article is to describe the Whole Brain Creativity Model and its implications for nursing education and practice. PMID- 11051931 TI - Adolescent interpersonal communication patterns. AB - Adolescents are admitted to psychiatric wards presenting with psychiatric problems which are essentially secondary to problematic interpersonal relationships. Successful interpersonal relationships however depend on effective interpersonal communication. Therefore the aim of research on adolescent interpersonal communication was to explore and describe the interpersonal communication patterns of adolescents and to develop an interpersonal communication skills approach to facilitate adolescent interpersonal communication skills within a training programme for adolescents. In this article however attention will be given to the description of the interpersonal communication patterns of adolescents. The target population of the research was 17 year old adolescents. The research consisted of a pre-phase where two contextual scenarios were formulated within group discussions with adolescents. During phase one of the research these scenarios were used to obtain video taped role plays from pairs of adolescents of the target population which were transcribed for data gathering purposes. Written dialogues were also obtained from each pair of adolescents on the same scenarios used for triangulation purposes. During phase two of the research the data was analysed according to Tesch's method and a literature control was done to verify the results. Guba's model for the trustworthiness of qualitative research was used. Four recurrent interpersonal communication patterns were identified, namely: Recurrent patterns of defocusing and externalizing the topic under discussion; Recurrent patterns of struggling for power; Recurrent patterns of not listening; and Recurrent patterns of focusing only on cognitive contents of messages and not on feelings. The research showed that adolescents have ineffective interpersonal communication patterns. Recommendations were made to facilitate adolescent interpersonal communication within an interpersonal communication skills approach. PMID- 11051933 TI - Perspectives of health workers regarding primary health care delivery to the rural population in the Bothaville district. AB - This article reports on the views of public health workers regarding recent changes in the delivery of primary health care to people living and working in the Bothaville rural area. These changes in mobile health care form part of the Initiative for Sub-District Support's programme to provide sustained, concerted support to sub-districts to bring about improvements in health care management and health care delivery. Main shortcomings of the recent changes were identified as inadequate transportation facilities in rural areas, insufficient information dissemination to rural dwellers and lack of farmers' participation in rural health matters. Furthermore, poor communication and co-operation between different public health services prevailed and the need for an integration of these services was emphasised. PMID- 11051934 TI - [The empowerment of nurses in health services--a conceptual framework]. AB - In a study of the empowerment of nurses in health services the conceptual framework served as departure in a survey of empowerment in health services in Gauteng. The aim with this part of the research methodology and the development of the conceptual framework was mainly to determine if theorists in the literature agree about the interpretation of the concept empowerment and the concepts underlined in the descriptions and definitions of the term "empowerment". PMID- 11051935 TI - The effect of the Maternal Care Manual of the Perinatal Education Programme on the attitude of midwives towards their work. AB - OBJECTIVE: In this study the changes in attitude of midwives towards their work following completion of the Maternal Care Manual of the Perinatal Education Programme (PEP), were determined. METHOD: A prospective, controlled trial was performed in a study, and two control towns in a region where PEP had not previously been used. All midwives caring for pregnant women in the three towns were included in the study. First the attitude of these midwives was determined by means of a questionnaire. Subsequent to this, the Maternal Care Manual was introduced and studied by the midwives in the study town. Following the completion of the Manual after 12 months, the attitude of all midwives was again evaluated using the same questionnaire. RESULTS: A total of 40 midwives in the study town and 53 in the two control towns were included in the study. There were no differences on comparing the ages of the midwives in the study town to those in the control towns. The attitude of the midwives in the study town improved significantly (p < 0.001). The mean result in the study town improved by 6.1 (24.4%) marks from 14.5 (58.0%) to 20.6 (82.4%). A significant shift also occurred in the range of the marks from 0-25 to 13-25. No changes were observed in the control towns. CONCLUSION: Most studies that have evaluated educational programmes measured improvement in health services, and did not evaluate changes in attitude. This study found that the attitude of midwives improved significantly in the study town. This positive attitude of midwives towards their work and their ability to perform their daily tasks must be an important component of any programme to improve the quality of care rendered to women during pregnancy, labour and the puerperium. PMID- 11051936 TI - Perceptions of postbasic nursing students in the use of seminars as a teaching method. AB - With the transformation of education in the country, and the outcomes-based education that is learner-centred fast gaining ground, nurse educators need to revisit the teaching methods and consider those that will foster reflective thinking through interactive constructing process. These methods, according to Cropley and Dave (1978:196) prepare learners for lifelong learning and to challenge problems in the working world. A descriptive and exploratory study using descriptive naive sketches to collect data from 44 final year post basic nursing students at a university was undertaken. The students volunteered to take part in the study. A content analysis according to Tesch's (in Cresswell 1994:55) method was used. A follow-up interview using an open-ended question to gather more in depth information as well as to validate the collected data was done with 10 students from the previous sample. Four positive and three negative categories and their sub-categories were identified namely: Positive categories:- encourage active participation, professional growth and maturity, facilitate reflective thinking, encourage facilitative communication and interpersonal skills. Negative categories were:- time consuming, causes conflict amongst students, inability to teach becomes an obstacle. A literature control was conducted to support and confirm the findings. Concluding remarks and recommendations for effective use of seminar as a teaching method are made. PMID- 11051937 TI - Informational and emotional support for cancer patients' relatives. AB - The purpose of this study was to find out what cancer patients' relatives think about the actions of health care professionals in terms of providing informational and emotional support. The sample consisted of 168 relatives of patients from oncological wards all over Finland. The data were collected with a questionnaire specifically developed for this study. Non-parametric tests were used for statistical analysis. The results clearly highlighted the importance of informing relatives about the patient's illness. However, less than one-third of the relatives said they received much information. Relatives' need for emotional support was clearly at a lower level than their need for information. Even so, only very little support was provided in relation to relatives' needs. PMID- 11051938 TI - Attitudes of cancer patients, their family members and health professionals toward active euthanasia. AB - This qualitative study describes the attitudes of four groups of people in cancer care toward active euthanasia. Patients (32) with incurable cancer, their family members (13), nurses (13) and physicians (13) participated in the study which was carried out in two central hospitals and in four health centres in Finland. The data was collected by means of focused interviews which were taped, transcribed and then analysed by content analysis. More than half of the participants said that they could ethically justify active euthanasia. Most of these were family members and nurses. The main reasons for their ethical justification were the terminal illness of the patient, the presence of suffering and pain and the patient's own request. Those who could not justify active euthanasia said that one human being has no right to decide death of another. Potential abuse, uncertainty about the finality of the situation, the possibility of effective alleviation of symptoms and the effects which the practice might have on medical staff were also mentioned by this group. The results of this study support the assumption given in the earlier literature that attitudes toward active euthanasia are most positive where terminally ill cancer patients are concerned. PMID- 11051939 TI - Oral care in cancer nursing. AB - The present study describes the perception of education, self-rated knowledge and attitudes towards oral care, performed oral care and co-operation with dentistry among nurses and enrolled nurses in charge of patients with haematological malignancies, lung cancer and head and neck cancer. A total of 137 nurses and enrolled nurses participated in a semi-structured interview based on a 43-item questionnaire. Nurses had less education in oral care and rated their knowledge on oral care lower than did enrolled nurses. Both groups reported a need for continuing education. Knowledge in oral care was rated to be poorest concerning oral status, oral signs and symptoms and fluorides. Nurses gave out information on oral complications and instruction in oral hygiene to a greater extent and examined the oral cavity more often than did enrolled nurses. A total of 18% felt uncomfortable in discussing oral hygiene with the patients and 45% objected to examining the oral cavity and stated patient integrity as the main reason. A majority reported that they received sufficient help from dentistry. There is a need for continuing education in oral care among nurses and enrolled nurses, which must be a responsibility of and in co-operation with dentistry. PMID- 11051940 TI - A cultural change in cancer education and training. AB - Collaborative education and training is the only way forward for the cancer care work force. The Calman-Hine report has become the benchmark on which to build practice since its launch in 1995. A forward looking group--the Cancer Care Alliance of Teesside, South Durham and North Yorkshire Education Group and the University of Teesside--adopted its principles from the Calman-Hine Report and developed some strategic projects to address and achieve them. A needs assessment of cancer education influenced the agenda within the Cancer Care Alliance. The education portfolio that was developed had a multiprofessional focus, incorporating both academic and non-academic study. PMID- 11051941 TI - A competency-based tool for education. AB - Through a multidisciplinary team approach, a competency-based tool has been developed to help to assess and identify training and development needs for staff working within the cancer field. The tool assists both individuals and organisations to gain a micro and macro picture of staff's existing skills and their development needs. PMID- 11051942 TI - Relating information-needs to the cancer experience. 1. Jenny's story: a cancer narrative. AB - This paper is based on a phenomenological study that used narratives to explore lived cancer experiences. The aim of the study was to determine the important issues for people with cancer that arose out of their cancer experience, and to place their information-needs within the stages of the cancer trajectory. The literature highlights the importance of information-giving; however, many problems are encountered with its provision. People with cancer frequently express dissatisfaction with the information given to them and experience difficulty in retaining and processing information. Six individuals were invited to tell the story of their cancer experience through in-depth interviews and narrative analysis uncovered thematic aspects of the lived experience. One interview in particular stood out as capturing the essence of a lived experience. Jenny's narrative had a beginning, a middle and an end, features that are traditionally associated with stories. This paper focuses on her story in depth, and illustrates the extent to which cancer can impinge on normal coping mechanisms. A diagnosis of cancer cannot be isolated from the other events in an individual's life, and themes emerged which showed that cancer impacts on different aspects of an individual's self-identity, including body image, family, social and work relationships. The cancer experience invariably begins before the point of diagnosis and information-needs clearly change over time. PMID- 11051943 TI - Relating information needs to the cancer experience. 2. Themes from six cancer narratives. AB - Many different issues may arise for individuals with cancer, where the provision of information can be an effective coping strategy. It is also clear that information needs change over time and vary from person to person. This paper considers six cancer narratives from a study seeking to identify the information needs of people with cancer that emerged out of their cancer experience. Six respondents were invited to tell their story through in-depth interviews and narrative analysis uncovered thematic aspects of the lived experience. Themes emerged which showed that cancer impacts on different aspects of an individual's self-identity, including body image, family, social and work relationships. Cancer was viewed as an intrusion and the illness engendered feelings of vulnerability that impacted on their normal coping mechanisms. This resulted in a decreased ability to process information. While individuals expressed medical information needs, they were less likely to articulate their need for information when it related to other areas of their lives. Individuals reached a turning point during their experience, when the self-acknowledgement that they were living with cancer, enabled them to become more active respondents in the information process. As this stage cannot readily be identified as occurring at a specific point of the cancer trajectory, communication channels need to be kept open regarding information-giving. This raises questions about areas for further study. PMID- 11051944 TI - What is FACET. Forum for Applied Cancer Education and Training. PMID- 11051945 TI - Palliative care issues in sub-Saharan Africa. PMID- 11051947 TI - District nurses' needs: palliative care for people with learning disabilities. AB - The aim of this article is to identify national palliative care issues, and current palliative care services provided by district nurses for people with a learning disability in North Staffordshire, UK. The professional and educational needs of district nurses in the role of palliative care providers for people with a learning disability were identified by two small focus groups involving district nurses. This resulted in the formulation of a questionnaire, which was circulated to 165 district nurses. While the initial part of the research is qualitative, the final survey involved a quantitative analysis using a questionnaire and a three-point Likert scale. This compared a number of independent variables such as age range, length of experience and number of contacts with clients who have a learning disability and palliative care needs. The results provide an overview of existing service uptake and a profile of the professional and educational needs of district nurses when caring for people with a learning disability who also have palliative care needs. PMID- 11051946 TI - A randomized trial of winged Vialon cannulae and metal butterfly needles. AB - The purpose of this parallel randomized study was to evaluate whether subcutaneous infusion sites initiated with winged vialon cannulae would have fewer skin reactions and longer site duration than metal butterfly needles and reduce needlestick injuries to staff. Data were collected on 42 hospice inpatients from the time of insertion of the first needle or cannula to the time of the first replacement of that device. A non-parametric survival plot for time (Kaplan-Meier method) was conducted and the survival time to replacement of the Vialon cannula was found to be longer than the metal butterfly needle. During the study period there were a total of four needlestick injuries to staff from metal butterfly needles. The authors conclude that Vialon cannulae reduce the frequency of site changes and are safer for staff to use because of the reduced risk of needlestick injuries. PMID- 11051948 TI - Rehabilitation and specialist palliative care. AB - Enabling individuals to achieve their maximum potential and quality of life following a diagnosis of advanced cancer, has long been a stated aim of palliative care. Increased life expectancy and the introduction of specialist palliative care to patients at an earlier stage of their illness presents professionals in the specialty with new challenges in meeting the need for rehabilitative care. This article examines some of the recent developments affecting the provision of rehabilitative care and describes one specialist palliative care unit's response to the challenge, highlighting the role of a nurse-led clinic within the service framework. PMID- 11051949 TI - Creutzfeldt-Jakob disease: extending palliative care nursing knowledge. AB - In a 12-month period six people died from Creutzfeldt-Jakob disease (CJD) in a Melbourne hospice. CJD is a rare neurodegenerative disease, which commonly follows an explosive course unabated into the terminal phase. For the purposes of this article the care of six patients was audited using a retrospective chart review and a focus group was conducted with nurses involved in their care. The nurses faced considerable challenges as they endeavoured to provide comfort and support for these patients. The differences in the illness trajectory of CJD in relation to the more common experiences of illness progression in advanced cancer are discussed in the context of palliative care. This review focuses on the particular care issues of six people with CJD and their families at the end of life. The personal issues experienced by the nurses who provided palliative care are also explored. The need for the future development of guidelines for families and health professionals who care for people with CJD is highlighted. PMID- 11051950 TI - Evaluating a palliative care education project in nursing homes. AB - This article reports on an evaluation of a 2-year palliative care education project for nursing home staff. The aim of the project was to provide education for all levels of nursing home staff so that the care of dying residents could be improved. In order to ascertain the outcomes of the initiative two approaches to data collection were adopted. Case studies of four participating nursing homes were undertaken, involving a period of participant observation and interviews with staff members. A postal survey of the participating nursing homes and non participating nursing homes from the same geographical region was also carried out. The impact of the project is described, identifying how the care of residents and their relatives was affected, the differing impact on the participating staff groups and the degree to which the organizational practices of the nursing homes changed. Although the project influenced course members' practice, the provision of courses such as these was recognized to be insufficient to ensure widespread organizational changes. Questions regarding the effectiveness of education as an agent of organizational change were raised throughout the project and the evaluation. PMID- 11051951 TI - Implications of the Stephen Lawrence Inquiry for palliative care. PMID- 11051952 TI - Breast cancer: net gains for patients. PMID- 11051953 TI - Professional development for professional developers. PMID- 11051954 TI - On being a person of integrity ... or ethics and other liabilities. AB - The emergence of managed care and the health industry's response to it has precipitated a moral crisis for many health care professionals. Caught between the patients' best interests and the employers' and payers' restrictions, professionals find their good intentions tested. In this article, the author distinguishes moral from ethical problems and suggests that self-examination and self-diagnosis of the mindsets and conditions that contribute to moral malfunctioning in the workplace--self-management at the individual level--is essential for maintaining the ethical stance of the profession and the integrity of the professional. PMID- 11051955 TI - Continuous professional development. AB - BACKGROUND: Lifelong learning is a requirement for all nurses. Structured continuing education programs are just one method of acquiring knowledge. Non structured, self-directed methods of development often are overlooked in this process. METHOD: This article presents suggestions for self-assessment and non structured methods of professional development for professional developers. RESULTS/CONCLUSION: By following the suggestions presented in this article, professional developers can assess their true learning needs and identify useful methods to develop themselves professionally. PMID- 11051956 TI - POTS, PANS, and PEARLs: the nursing profession's rich history with distance education for a new century of nursing. AB - BACKGROUND: Continuing education via distance education is expanding. A review of the media used to deliver classes in the past as well as future possibilities are presented. METHOD: The personal experience of the authors using these methods to present continuing nursing education is the basis for this article. RESULTS: There is evidence that people learn via distance education as well as they do with traditional formats such as conferences and seminars. Although learners are enrolling in these types of courses for convenience, they enjoy the classes more when there is active involvement and participation. CONCLUSIONS: Continuing education using distance education strategies will expand, especially because computers are available in the homes of learners. A larger number of offerings will be available, and more learners will use these media to remain current professionally. PMID- 11051957 TI - Working smart: turning everyday commitments into scholarly outcomes. AB - BACKGROUND: This article offers a model for transforming routine professional responsibilities into scholarly products. METHOD: Using Boyer's model of scholarship as a guide, the authors were able to transform a professional commitment into the scholarship of teaching and learning, practice and engagement, and discovery. RESULTS: The authors were able to conduct a research study, educate graduate students, and provide important data about nursing to a statewide project. CONCLUSION: Working smart requires transforming existing commitments and resources into opportunities for creating scholarly products that are shared with the broader nursing community. PMID- 11051959 TI - Perspectives on faculty development. AB - This article reviews the requirements necessary to address nursing faculty development needs to continue to strengthen nursing education for state-of-the art reflective practice. Included in the discussion are the importance of information and sophisticated information management; strategies to enhance teaching competency, efficiency, and the quality of instruction; current measures of quality facilities and resources; and the contributions of teaching and learning centers. The discussion ends by emphasizing the need to embrace the concept of differentiated nursing practice. PMID- 11051958 TI - Managing scholarship in the faculty role. AB - BACKGROUND: This article was written to describe approaches the author used that ultimately resulted in tenure and promotion. METHOD: Self-reflection was used to identify the progressive steps that lead to being tenured and promoted. RESULTS: This article describes the key factors to remember in gaining promotion and tenured status. Personal experiences and life philosophy describe the essence of the steps taken. CONCLUSION: Achieving tenure and promotion, while never easy, is best accomplished with a clear vision of the desired outcomes, a plan that allows incorporation of alternative opportunities, willingness to persevere in the face of obstacles, and the ability to see humor in the midst of turmoil. PMID- 11051960 TI - Professionalists, all. PMID- 11051961 TI - Taking the pain out of waiting. PMID- 11051962 TI - A comparative study of transformational leadership in nursing development units and conventional clinical settings. AB - AIMS: This is a comparative study of the leadership provided by nurse managers and leaders in Nursing Development Units and conventional clinical settings in England. BACKGROUND: Nursing development units (NDUs) were originally conceived as centres of nursing excellence, innovation and leadership development. This article describes the first published use of a leadership practices inventory (LPI) explicitly based upon a model of transformational leadership. This style of leadership has been commended as highly effective and suitable for nursing. METHODS: The use of the LPI was piloted as a postal questionnaire and as a schedule for telephone interviewing, these pilots supported the use of telephone interviewing in the main study. Two matched samples of 70 nurses in total were recruited from across England, comprising 14 nurse leaders and 56 of their day to day colleagues. Data was collected by telephone interviewing over a 6-week period between February and April 1998. Six null hypotheses were developed to identify significant inter-group differences in leadership behaviour. Descriptive and inferential data analysis techniques were employed using SPSS for Windows. FINDINGS: The leadership provided by NDU leaders was evaluated more highly than non-NDU leaders. A higher level of congruence between self and observer evaluations was shown by NDU leaders. Statistically significant inter-group differences were apparent in three of the five practices of exemplary leadership and in the overall leadership behaviour. NDU leaders show greater self awareness and are more transformational than their non-NDU counterparts. The limitations of the study design are discussed. CONCLUSIONS: NDU leaders provide leadership of a more transformational nature than their counterparts working in conventional settings. This finding suggests that NDU leaders have enhanced leadership potential and that formalizing nursing development within NDUs may promote the emergence of transformational leadership and provide a microculture in which it might flourish. The LPI is regarded as a useful, adaptable tool suitable for use in UK nursing applications including research, leadership development and education. PMID- 11051963 TI - Shared governance--nurses making a difference. AB - AIM: This paper aims to describe how shared governance can be successfully integrated into existing management structures in a large medical directorate. It will show how the shared governance philosophy can lead to the creation of a culture where nurses feel important and valued and also consider its use as a foundation for the implementation of the nursing strategy. BACKGROUND: The hospital adopted shared governance in 1995 with the setting up of councils led by staff nurses. Shared governance advocates claim it broadens involvement of clinically-based nurses in the process of change through devolved decision-making thereby enhancing clinical practice and increasing staff moral and self-esteem. Key issues Shared governance is a cultural change that will develop the leadership and management skills of all grades of staff. Shared governance is not a quick fix for the profession; involvement of all staff needs time, persistence, determination and a strong commitment to training and development. CONCLUSION: The national nursing strategy puts nurses in a position to positively influence health care for the advantage of patients, however, for nurses to grasp this opportunity cultural change is required. This practical example of shared governance demonstrates how it can be used to create a proactive culture, focused on improving patient care. The nurturing and developing of clinically-based nurses provides them with the knowledge and skills to challenge the status quo and lead change. Thus, shared governance is an excellent foundation from which to develop the nursing strategy. PMID- 11051964 TI - A nursing theory for nursing leadership. AB - For many years nursing practice has found its foundations in nursing theory. A review of theorists such as D. E. Orem, C. Roy, B. Neumen, V. Henderson, M. E. Rogers and others reveals a focus on the management of patient care, not leadership. This has provided most nurses with a solid foundation in 'management', but little in terms of 'leadership.' In more recent years, theories such as the Deming Management Method, Managers as Developer Model, Shared Governance and Transactional Leadership have been introduced, none of which are nursing theories. This article discusses the conceptualized differences between management and leadership theory arguing that there is a difference between 'leadership and management'. A leadership theory is proposed utilizing Ida J. Orlando's model for nursing. This theory provides a nursing foundation for nursing leaders to utilize both in the management of patient care and in leadership. PMID- 11051965 TI - Approaches to preventing burnout: the effects of personal stress management and organizational socialization. AB - Several approaches to preventing burnout are compared. One hundred and fifty-four nurses in five Hong Kong hospitals completed the Maslach Burnout Inventory (MBI), the Organizational Socialization Inventory (OSI), and three measures of personal stress management. Results indicated that favourable evaluations on the four OSI domains (job training, organizational understanding, coworker support and future prospects) yielded strong negative correlations with the burnout components. Also, the personal stress management measures had strong negative correlations with depersonalization and decreased personal accomplishment, but none were related to emotional exhaustion. Stepwise regression analyses indicated that training was the only (inverse) predictor of emotional exhaustion, whereas interpersonal skills and understanding were strong (inverse) predictors of depersonalization. Additionally, interpersonal skills and coworker support were excellent (inverse) predictors of decreased personal accomplishment. The findings are discussed in terms of their relevance to nursing administration. PMID- 11051966 TI - Value of admission interviews in selecting of undergraduate nursing students. AB - METHODS: The attrition of students in the undergraduate nursing program was evaluated. The value of the admission interview and its preferred nature were assessed by questionnaires filled by candidates and faculty. FINDINGS: It was found that although interviewing was only partially effective in screening potential drop-outs, including those who left the programme on non-academic grounds, without interviews the attrition rate increased. Personal interviews accounted for a somewhat lower attrition rate than the unanimously preferred group interviews. CONCLUSION: The process of choosing candidates to nursing programmes requires new thinking, as high attrition rates are not prevented by interviews. More research into the proper mechanism of student selection is needed. PMID- 11051967 TI - Patients' perceptions of a pre-admission clinic. AB - AIM: Patients' perceptions of a pre-admission clinic in an orthopaedic hospital were explored. The clinic was developed as a collaborative initiative between nursing and medical staff. METHODS: The methodological approach to this study was inductive and informed by grounded theory. Data was collected using unstructured interviews and analysed using constant comparative analysis. RESULTS: The clinic appeared to relieve patients' anxiety, however, the way in which it did so was complex. Patients thought they were given factual information relating to surgery in the clinic and this was conceptualized as 'given information'. Patients also told of certain experiences in the clinic, which included the caring environment, individualized care and efficiency. These experiences were powerful tools in giving information to patients and are conceptualized as 'experienced information'. CONCLUSION: Both 'experienced information' and 'given information' were important in relieving anxiety. Coping styles of patients differed. Individual patients selected from the clinic what they found useful in terms of 'given information' and 'experienced information' in accordance with their individual coping styles. PMID- 11051968 TI - Enrolled nurse conversion: a review of the literature. AB - AIM: The aim of this literature review was to examine the policies and professional literature from the last 50 years about the introduction, the role and subsequent plight of the enrolled nurse (also known as second level nurses), and the need to convert to the first level of the UKCC nursing register. BACKGROUND: Nurse shortages within the NHS have been cyclical since its inception in 1948. The policy decision to cease the training of enrolled nurses within the frame of modernizing the education and training of the nursing workforce had two distinct implications for enrolled nurses. Firstly, they could choose to stay as enrolled nurses or convert to first level nursing. Nevertheless, enrolled nurses have cited the lack of funded conversion course places, and managerial support for non-conversion. METHODS: A critical review of the national policies and professional literature concerned with the evaluation of enrolled nurses' contribution to the NHS. FINDINGS: It was argued that national policy needs to be supported on the ground, whereby enrolled nurses are proactively supported to come forward for conversion and/or meaningful roles are created and sustained where enrolled nurses continue to make a valuable contribution to the NHS agenda. Finally, the paper challenges all NHS organizations to consider the profile and value of enrolled nurses and become proactive in their recruitment and retention of this nursing group. PMID- 11051969 TI - President's message. Improving the prospects of the world's 6 billionth child. PMID- 11051970 TI - Freedom. PMID- 11051971 TI - Eye bruising. PMID- 11051972 TI - Working as a locum tenems midwife. PMID- 11051973 TI - My top 10 favorite complementary modalities. PMID- 11051974 TI - Touching lives. Massage in pregnancy and labor. PMID- 11051975 TI - Providing choice: naturopathic midwifery. PMID- 11051976 TI - Bodily knowledge: the wisdom of nausea. PMID- 11051977 TI - Chiropractic care in pregnancy. PMID- 11051978 TI - Midwives as the quintessential barefoot doctors. PMID- 11051979 TI - An unassisted homebirth. PMID- 11051980 TI - Why home is best. PMID- 11051981 TI - Hospital birth: a CNM philosophy. PMID- 11051982 TI - Vegan pregnancy. PMID- 11051983 TI - Blue cohosh and newborn myocardial infarction? PMID- 11051984 TI - Stay in the humble: a double footling breech birth. PMID- 11051985 TI - Evidence-informed midwifery. 2. Using research in midwifery practice. PMID- 11051986 TI - Stay calm with PROM. PMID- 11051987 TI - Group B strep screening. PMID- 11051988 TI - Open house: Email lists for midwives, douglas and other birth practitioners. PMID- 11051989 TI - My road to midwifery. PMID- 11051990 TI - Home waterbirth in England. PMID- 11051991 TI - Hawa Gola and mother-in-law's big toe. PMID- 11051992 TI - School nurses: insuring children's health. PMID- 11051993 TI - National task force on confidential student health information offers guidelines. PMID- 11051994 TI - A fairy tale come true? PMID- 11051995 TI - Attention deficit hyperactivity disorder: experts on call. PMID- 11051996 TI - Specialized educational opportunities for school nurses. PMID- 11051997 TI - Asthma: promoting best practice in the school setting. PMID- 11051998 TI - Collaborative off-shore nurse education: an Australian initiative. PMID- 11051999 TI - Performance assessment. PMID- 11052000 TI - Authoring software for courses delivered on the Web, Part 2: WebCT. PMID- 11052001 TI - Developing student evaluation instruments to measure instructor effectiveness. AB - The authors detail the process of developing four specific tools for student evaluation of faculty performance at the master's and baccalaureate levels. To accomplish the desired goal of improving instruction, these authors believe student evaluation tools should be tailor-made to the educational setting and mode of delivery. The tools described in this article were developed to evaluate faculty performance in clinical-direct, on-campus laboratory, clinically precepted, and didactic settings. PMID- 11052002 TI - Portfolios: documenting learning in a personal way. PMID- 11052003 TI - Infusing critical thinking into the nursing curriculum through faculty development. AB - An important prerequisite to changes in an educational program's predominant philosophy and culture is systematic faculty development to begin and sustain the evolution. The authors describe how one nursing school implemented a faculty development program with the goal of infusing critical thinking strategies into courses throughout the curriculum. PMID- 11052004 TI - Support for breastfeeding students. PMID- 11052005 TI - Concept mapping: reducing clinical care plan paperwork and increasing learning. AB - The author describes how concept maps were used in place of nursing care plans to reduce care planning paperwork in fundamentals and medical-surgical clinical courses in acute care facilities. In addition to less paperwork, clinical concept mapping enhances students' critical thinking skills and clinical reasoning because students and faculty can clearly and succinctly visualize priorities and identify relationships in clinical patient data. PMID- 11052007 TI - Teaching case management: a case study approach. PMID- 11052006 TI - The value of student portfolios to evaluate undergraduate nursing programs. AB - A growing trend in nursing education is the use of student portfolios for program evaluation. Incorporating portfolio analysis into a school's evaluation plan requires that faculty consider how the benefits and limitations of the portfolio development process impact the entire curriculum. The primary benefit of portfolio evaluation is that it permits the correlation of competencies attained by graduates with curricular outcomes. However, portfolio development also promotes increased student responsibility for learning, enhances faculty-student interaction, and facilitates changes in curriculum and instruction. The principal limitations of portfolio evaluation are the lack of research-based evidence that demonstrates the validity and reliability of portfolio analysis, time required to create a portfolio, and documentation storage. PMID- 11052008 TI - International collaborative workshops. A 6-year partnership between Canada and China. AB - Interprofessional exchanges such as workshops are common activities in international development programs. The authors describe processes used to select, implement, and evaluate six collaborative workshops held in Tianjin, China, between 1990 and 1995. These workshops targeted curricular change for a baccalaureate nursing program in Tianjin Medical University. Our experience highlights the importance of understanding the assumptive base for change, developing an institutional memory in both partner universities, introducing interactive teaching methods congruent with Chinese value systems, and using a formative evaluation strategy to foster cross-cultural dialogue and mutual understanding among project partners. PMID- 11052009 TI - Conceptual frameworks. Putting the nursing focus into core curricula. AB - The growing movement toward specialty certification in nursing has resulted in an increased demand for continuing education that supports specific areas of specialty practice and certification status. In examining a number of the specialty practice core curricula documents, it is evident that there is a tendency to organize nursing content using the medical model as a framework. A method by which core curricula for specialty practice are organized and developed from a distinctly nursing perspective is needed. The authors suggest that conceptual frameworks, developed by nurses who are expert in a specific clinical practice area, are a practical method for the development of such curricula. PMID- 11052010 TI - Integrating political involvement and nursing education. AB - Numerous nursing leadership organizations and nursing experts emphasize the need for nurses to develop health policy skills. However, most nurses continue to lack this skill. Few examples of successful ways to deal with health policy in the curriculum can be found in the literature. The authors present a discussion of how health policy skills may be developed at various curriculum levels and use a Master's core course in health policy as an example of a graduate course. PMID- 11052011 TI - Reality checks. PMID- 11052012 TI - Why are shortages of hospital RNs concentrated in specialty care units? AB - This article is the first in a series examining the interplay between the aging of the nurse workforce and other factors driving the growing nursing shortage that are already affecting some specialty areas. Nearly 60% of the current RN workforce is over 40 years of age; and the percentage of RNs under age 30 has fallen by nearly 40% since 1980. The total number of FTE RNs is projected to shrink after 2010, likely resulting in shortages of RNs "when the large baby-boom generation of RNs starts to retire." Because ICUs have historically attracted younger RNs, the rapid decline in the number of RNs in the workforce under age 30 plays a large role in explaining the development of shortages in the ICU. The growing difficulties staffing operating rooms and other peri-operative services is seen as related to the aging work force as more diploma prepared nurses have been attracted to this specialty because they had educational exposure to this area. PMID- 11052013 TI - Telephone nursing: evidence of client and organizational benefits. AB - Telephone nursing (TN) or telephone triage (TT) has been identified as part of a successful cost-reduction demand management strategy. The author examines TN utilization and related client satisfaction, client education, reduction in drop in clinic visits, and unnecessary ER and urgent care visits associated with an outpatient pediatric clinic population. This study examined 25% of the total of an average of TN 120 phone calls processed by this Southwestern clinic in an average summer month and achieved an 87.3% response rate to followup study questions. "Telephone nursing was performed by specially trained and experienced RNs using approved, written, clinic-specific protocols." The primary goals of the TN program was the "efficient use of health care resources" and provision of the "appropriate level of care at the appropriate time." Over 80% of the callers surveyed reported that if they hadn't been able to speak to the nurse they would have sought medical care elsewhere. PMID- 11052015 TI - DONs in long-term care facilities: contemporary roles, current credentials, and educational needs. AB - The need for educated nursing leaders in long-term care facilities is critical now and will continue to be well into this century. The purpose of this research was to replicate and geographically extend a descriptive survey conducted by Heine (1995) to determine the roles and responsibilities of directors of nursing (DONs) in long-term care nursing facilities (LTCNF) as well as their educational preparation, current professional credentials, and educational needs. 945 DONs in LTCNF in New England (Connecticut, Maine, Massachusetts, New Hampshire, Rhode Island, and Vermont) as identified in state directories of nursing home facilities were surveyed. Completed questionnaires were returned by 247 DONs for an overall response rate of 26%. DONs in LTCNF reported that they were most involved in roles/responsibilities related to nursing/health services management and least involved in professional nursing and long-term care management. PMID- 11052014 TI - Nurse staffing decisions: an integrative review of the literature. AB - The author exhaustively explores the current literature and attempts to summarize the current thinking on how to best decide on the most cost-effective nurse staffing requirements. Between 1984 and 1994 FTE nursing employees decreased by 7.3%, causing some researchers to seek ways to explore the relationship between staffing levels, staff and patient satisfaction and outcomes of care. Satisfaction among staff nurses working in a self-scheduling environment was determined largely by the individual's ability to negotiate for the desired days and shifts and by the nurse manager's ability to stand back from the process and let the staff collaboratively work it out. Work structure related studies seemed to find that 12-hour shifts were reported to be "less fatiguing" than traditional 8-hour shifts. Staffing studies found that rural hospitals still used 0.27 more RNs per occupied bed than urban hospitals and that the presence of a unit secretary was associated with a decreased use of RNs. PMID- 11052016 TI - Balancing from the balcony. PMID- 11052017 TI - I'm behind you! The manager as a coach. PMID- 11052018 TI - Integrated delivery systems: Mercy Health Services, Part II. Interview by SueEllen Pinkerton. PMID- 11052019 TI - Working with a board. AB - An effective board and an effective organization go hand-in-hand. It is worth the time of the chief executive or senior staff to develop relations with the board. Participating together in board development programs can be valuable for both. A strong partnership between board and staff members promotes organizational commitment and success. PMID- 11052020 TI - Medicare, Social Security, and competitive benefits are neglected nursing issues. PMID- 11052021 TI - Organising antenatal education for safe motherhood. PMID- 11052022 TI - The Ilizarov fixator. PMID- 11052023 TI - Nurses' role in occupational health services. PMID- 11052024 TI - Flow sheet for better documentation in the labour room. PMID- 11052025 TI - Just when you thought you knew it all... PMID- 11052026 TI - Latex allergy. An orthopaedic case presentation and considerations in patient care. AB - Latex allergy has become a challenging phenomenon in health care delivery for the last several years given the numerous products containing latex. The number of individuals with latex allergy has grown dramatically since the institution of standard precautions issued by the Centers for Disease Control and Prevention (CDC) in 1987 in response to the AIDS epidemic. The allergic reaction to latex ranges from a minor skin rash to anaphylactic shock. Preventing exposure to latex is the key to managing and preventing this allergy. Providing a safe environment for patients with latex allergy is the responsibility of all health care professionals. Identification of patients with such an allergy or those at risk is the initial step toward providing a safe environment. Care givers must have a clear understanding of the routes of exposure, effective prevention modalities, the types of allergic reactions that may be manifested by latex sensitivities, the signs and symptoms of allergic reactions, and the treatment options related to the reactions. An orthopaedic case presentation represents a multidisciplinary team approach that successfully managed a patient undergoing a total knee arthroplasty. PMID- 11052027 TI - Supracondylar fractures in children. AB - Musculoskeletal trauma accounts for approximately 15% of all childhood injuries (Devito, 1996). Fractures are a common injury sustained with trauma. Due to anatomic, biomechanical, and physiologic differences between adult and pediatric bones, patterns of fractures change as a child's age increases. The treatment of a pediatric fracture may also differ from that of an adult due to these differences. This article discusses one common type of pediatric fracture, the supracondylar fracture. The etiology, incidence, classification system, and treatment of this fracture will be explained. In addition, potential complications and nursing management of a supracondylar fracture are included. PMID- 11052028 TI - Children with sickle cell anemia and orthopaedic problems. AB - Among the many unique challenges facing orthopaedic nursing is the care of children with sickle cell disease. Whether the condition that initiates the contact is related to the sickle cell disease or is incidental to it, nurses need a basic understanding of the disease process to adequately help these patients reach healthier outcomes. Sickle cell disease is an autosomal recessive inherited condition seen primarily in the African-American population. Orthopaedic nursing care for these patients must incorporate multisystem, developmentally appropriate assessments and interventions. Pain control, hydration, establishment of trust, and support of individual and family coping all play vital roles in insuring a rapid recovery and a reduction in long-term complications. PMID- 11052029 TI - Surgical hip dressings: a comparison of taping methods. AB - PURPOSE: To examine the effectiveness of two methods of preventing blister formation under the taped portion of postoperative hip surgical dressings. DESIGN: Comparative intervention study. SAMPLE: Convenience sample of 148 consecutive hip surgery patients in two Connecticut community hospitals. METHODS: Patients were preoperatively assigned to one of two postoperative surgical hip dressing taping methods or to the control group. The experimental group had the surgical dressing taped to either a hydrocolloid barrier or a nonhydrocolloid barrier 1-inch circumferentially around the surgical incision with the control group having the dressing taped directly to the skin. FINDINGS: The research results found that taping the surgical dressing to a hydrocolloid barrier prevented blister formation. There were no blisters in either experimental group, but evidence of nonblanchable erythema on two patients in the nonhydrocolloid barrier group. CONCLUSION: Taping of postoperative surgical hip dressings to a hydrocolloid barrier is superior to taping directly to skin or to a nonhydrocolloid barrier. IMPLICATIONS FOR NURSING RESEARCH: A larger scale study to examine potential factors that might place patients at high risk for blister formation is needed. This would aid in identification of patients that would benefit from different surgical dressing taping methods. PMID- 11052030 TI - Wound complications after hip surgery using a tapeless compressive support. AB - PURPOSE: To compare the incidence of wound complications after hip surgery in patients treated with a compressive spica wrap dressing to those using traditional taping methods. DESIGN: Retrospective, descriptive, comparative, intervention study. SAMPLE: 457 hip surgery patients, including primary arthroplasty, revision surgery and fracture with ORIF. METHOD: A compressive wrap dressing was used on hip surgery patients in the study group while tape was used on patients in the control group. A retrospective chart review was conducted noting postoperative wound complications. FINDINGS: There was a significantly lower incidence of blisters and drainage in the study group using a compressive wrap dressing. There was not a higher incidence of DVT or infection using the wrap dressing when compared to published studies. CONCLUSION: Use of a compressive wrap dressing after hip surgery is recommended to reduce the risk of wound complications. IMPLICATIONS FOR NURSING RESEARCH: A prospective, randomized study with multiple surgeons using both compressive wraps and traditional taping techniques would substantiate the advantage of using a hip spica dressing after hip surgery. PMID- 11052031 TI - Transdermal drug delivery using iontophoresis and phonophoresis. AB - Introducing medicines into the human body by way of the skin is an ancient practice, and transdermal delivery has long been a standard for administering medications such as nitroglycerin and scopalamine. Phoresis, another method of transdermal drug delivery, is now being ordered for an increasing number of orthopaedic patients who suffer from inflammation, strains, or sprains. In phoretic drug delivery, enhancers such as electricity or ultrasound are used to stimulate drug absorption in the treatment area. To guide their patients to explore a variety of treatment options, orthopaedic nurses need a greater understanding of these phoretic modalities. PMID- 11052032 TI - Synovial osteochondomatosis. AB - Synovial osteochondomatosis is a benign, metaplastic disease of unknown etiology in which synovial tissue can differentiate into cartilage and/or bone. This cartilage and bony tissue may be attached to the synovium or may break free and become intraarticular loose bodies. The treatment of choice for this unusual condition is synovectomy, which is usually curative. PMID- 11052033 TI - An orthopaedic workshop and skills laboratory. AB - Planning and implementing care for patients with musculoskeletal disorders can be intimidating for nursing students. Moreover, changes in health care delivery often limit the opportunities students will have to learn skills and participate in the care of a patient in need of these services. This article describes the development of an orthopaedic workshop and skills laboratory which provides hands on learning experience in a nonthreatening environment. The teaching strategy uses the collaborative resources of health care providers, allowing the student to be introduced to the care of orthopaedic patients from admission to discharge. PMID- 11052034 TI - Neurovascular assessment. AB - The nurse who is thoroughly familiar with the components and underlying scientific rationale of the neurovascular assessment can perform the assessment capably, competently, and quickly. It is the nurse's responsibility to perform neurovascular assessment, document accurate patient data, and communicate the assessment findings to other members of the health care team in order to prevent or promptly detect signs and symptoms of potential or developing complications. PMID- 11052035 TI - Introducing ... Carolyn Crane Cutilli. PMID- 11052036 TI - Keeping them safe. PMID- 11052037 TI - Prevention: the only cure for pediatric trauma. AB - In this age of modern medicine, one would like to believe that we have come far, but it is not so with pediatric trauma. Statistics remain frighteningly high with trauma as the leading cause of death in children. It affects 1 in every 3-4 children. Up to 22 million children suffer injuries every year. In 30 years, the statistic of 50% of all childhood deaths resulting from injuries has remained unchanged, but the number of injured children continues to climb. The cost to society is in the billions. Nurses can decrease morbidity and mortality in children by understanding common patterns of injury and educating parents and children about injury prevention. PMID- 11052039 TI - Substance abuse and the hospitalized elderly. AB - Health care professionals frequently fail to recognize and address the misuse and abuse of alcohol and drugs in the elderly. Estimates of alcohol abuse in the older adult population range from 4% to 20% in the community dwelling elderly and up to approximately 25% among hospitalized older adults (Adams & Cox, 1995; Adams & Kinney, 1995; Beresford et al., 1990). In addition, the present population of older adults consumes 2-3 times more psychoactive medications than younger age groups (Sheahan et al., 1995). The effects of alcohol and substance abuse in older adults are influenced by physical, developmental, and psychosocial changes that occur with aging. Identification of alcohol and substance abuse presents a challenge for health care providers as older adults often present with atypical symptoms. Accurate diagnosis allows for the initiation of interventions for both immediate and long-term treatment. PMID- 11052038 TI - Case study. Fractured ankle. AB - To improve our practice performance for patients admitted to the emergency department with fractured ankles, our Clinical Quality Improvement Team (CQI) developed the Fractured Ankle Critical Path, for patients needing open reduction and internal fixation, but for whom surgery could or should be deferred as determined by the orthopaedic surgeon. Members of the team included an orthopaedic trauma surgeon, an emergency department physician, case managers, and representatives of the Home Care Department at Rhode Island Hospital. The critical path includes a Home Care referral procedure, an R.I. Hospital Fracture and Orthopaedic Information Sheet, and a Fractured Ankle Home Care Protocol. The goal of the Fractured Ankle Critical Path is to decrease length of stay while maintaining positive outcomes. PMID- 11052040 TI - Improving pain management after total joint replacement surgery. AB - PURPOSE: To test the effects of implementing evidence-based postoperative pain management content and strategies on patient, provider (nurse and physician), and fiscal outcomes. SAMPLE: 118 patients, 57 before and 61 after implementation, having total knee replacement (TKR) (54%) and total hip replacement (THR) (45%), and 28 orthopaedic nurses. METHODS: A research utilization approach with a pretest/posttest design was used. Independent variables (interventions) included evidence-based pain management content, education of care providers and patients, and system changes at the point of care. Dependent variables (outcomes) were patient perception of the postoperative pain experience, provider practice patterns, and length of stay (LOS). FINDINGS: The hypotheses of decreased provider use of meperidine and increased use of hydromorphone, i.v. route, pain plans of care, and nurse knowledge were supported. LOS was significantly reduced. The patient hypotheses decreased pain intensity and side effects and increased satisfaction and function were not supported. CONCLUSION: Methodical implementation of evidence-based pain management information changed practice and fiscal outcomes. Improvement in the patient perception of pain management was more difficult to achieve. PMID- 11052041 TI - Congenital idiopathic clubfoot. AB - Clubfoot or talipes equinovarus is a complex deformity characterized by three distinct manifestations. The foot is in an equinus position; the forefoot and heel are in varus; and the entire foot is supinated. While the exact etiology of this problem remains unknown, many advancements have been made in the treatment of clubfeet. The surgical procedure is progressive and tailored according the severity of the deformity. Pain management, thorough neurovascular assessments, and education of the family are essential to caring for these patients. PMID- 11052042 TI - Caring for the orthopaedic patient receiving continuous ambulatory peritoneal dialysis. AB - Providing nursing care for patients with End stage renal disease (ESRD) receiving continuous ambulatory peritoneal dialysis (CAPD) provides a variety of challenges for the orthopaedic nurse. In ESRD the kidneys not only lose the ability to remove waste products and excess fluids, but also are unable to assist the body to use active vitamin D3 for calcium absorption. ESRD patients are at a high risk for developing fractures due to the kidneys' inability to excrete phosphate, resulting in elevated plasma phosphate and low plasma calcium levels resulting in osteodystrophy. Patients conduct peritoneal dialysis treatments independently after receiving an in-depth education program. It is important for the nurse to continue to foster patients' independence by encouraging self-administration of CAPD even during hospitalization. CAPD is carried out manually by instilling dialysis solution into the peritoneal cavity usually four times daily to remove excess fluids and metabolic waste products. Orthopaedic nurses face the challenge of not only caring for the patient's orthopaedic injury but also ensuring CAPD is implemented correctly. This article outlines common problems experienced by ESRD patients, principles of CAPD, and expected nursing care. PMID- 11052043 TI - Clinical evaluation of an automated turning bed. AB - PURPOSE: The purposes of this study were to assess client comfort and sleep quality, client physiologic response (skin and respiratory status), the effect on the need for caregiver assistance, and cost when using an automated turning bed. DESIGN: Nonexperimental, evaluative study. SAMPLE: Twenty-four adult home or long term care resident subjects who had a degenerative disease, spinal cord injury, stroke, cerebral palsy, or back surgery. METHODS: Each subject agreed to use the automated turning bed for four weeks. Researchers completed a demographic survey and skin assessment, and assessed each subject for pressure ulcer risk and for the need of assistance of a care giver for turning before and after the four weeks of using the turning bed. Subjects rated the turning bed in terms of comfort and sleep quality. FINDINGS: Subjects rated the turning bed as more comfortable than their own bed and expressed satisfaction at the pain relief attained when on the turning bed. While using the turning bed, there was a significant improvement in sleep quality. No skin breakdown or deterioration in respiratory status occurred. Fewer subjects required the assistance of a caregiver for turning when on the turning bed. CONCLUSION: This automated turning bed shows great promise in meeting a need for patients with limited mobility whether they are homebound or in a residential community. IMPLICATIONS FOR NURSING RESEARCH: Future studies that further investigate use of the turning bed for postoperative back patients while still in the acute care setting are indicated. Replicative studies with a larger sample size are also indicated. PMID- 11052044 TI - Managed care and critical pathway development: the joint replacement experience. AB - This article examines the economic, social, ethical, and political issues affecting total joint replacement patients in a managed care environment. Using general systems theory as a framework, it examines the interrelated historical events that have shaped the development of both joint replacement procedures and managed care, and discusses the extent to which these two phenomena have been mutually influential. Specifically, the article examines the initial development, implementation, and continuing evolution of clinical pathways as an easily identified and relatively discrete manifestation of managed care for the joint replacement population. While the overall impact of managed care is beyond the scope of this presentation, it is hoped that a focus on the practical application of clinical pathways to joint replacement will allow some general principles to emerge that may be useful for both patients and practitioners operating in other aspects of the managed care environment. PMID- 11052045 TI - The patient no one liked. AB - Despite professional admonitions to treat patients with dignity and to promote their well being, there are some patients who are unpopular with or not liked by nurses. Caring for these patients presents an ethical challenge for nurses. The purposes of this article are to explore the issue of bias and the use of labels, as well as the subsequent ethical concerns of respecting the uniqueness of the patient and preventing harm when nurses have a negative bias toward and label particular patients. The authors discuss several implications for nursing practice, including engaging in introspection, understanding the "problem" patient, actively involving patients in their care, and evaluating strategies used to eliminate negative bias and labeling. PMID- 11052046 TI - The upper extremity: traumatic injuries and conditions. PMID- 11052047 TI - Introducing ... Melinda Mock. PMID- 11052048 TI - Calcium intake in midlife women: one step in preventing osteoporosis. AB - Osteoporosis is widely viewed as a major health concern. It is a progressive bone disorder associated with pain, disability, and death secondary to its fracture complications. While more than 25 million Americans are afflicted, women succumb to its devastating consequences six to eight times more often than men. While many factors increase the risk of osteoporosis, attention must be directed to those measures that positively influence peak bone mass and allay bone loss. Minerals--particularly calcium--play a vital structural and mechanical role in bone growth and development. Moreover, an adequate intake of calcium is vital in the years surrounding menopause when bone loss accelerates. As nurse providers, our challenge resides with prevention strategies that aim at nutritional fitness for building and maintaining healthy bones. PMID- 11052049 TI - Osteoporosis diagnosis and fracture. AB - PURPOSE: The purpose of this study was to examine correlates for osteoporosis diagnosis and hip fracture among a national sample of women. DESIGN: Data were extracted from the Third National Health and Nutrition Examination Survey (NHANES III). This large-scale data set was collected by the National Center for Health Statistics. SAMPLE: The sample for this study included 2,336 women aged 50 years and older who resided in household interviewed for NHANES III. METHODS: Predictors for examination included age, race, heredity, body mass index, physical activity, smoking status, alcohol use, and dairy product use. Analyses were conducted using SAS procedures. FINDINGS: Correlates for screening and diagnosis of osteoporosis included age and race. Risk factors predicting hip fracture included age, race, low body mass index, and inactivity. IMPLICATIONS FOR NURSING PRACTICE: Recommendations emphasize screening of high-risk women, achieving and maintaining healthy body weights for underweight women, and obtaining moderate physical activity. PMID- 11052050 TI - The Jehovah's Witness orthopaedic trauma patient: an ethical challenge. AB - Members of the Jehovah's Witness faith believe that they are biblically prohibited from accepting blood transfusions even in cases of severe anemia that could lead to death. The refusal to accept blood is an ethical challenge for the health care team when managing an unstable Witness patient. The Jehovah's Witness patient has a right to refuse blood, and the health care team is obligated to abstain from providing transfusions. It is important that the health care team is proactive in becoming educated on the principles of the Jehovah's Witness faith and how to manage the patient without the administration of blood and blood products. The health care team is also encouraged to network with resources to provide support for the patient and family. PMID- 11052051 TI - Pediatric sedation: the art and science. AB - Successful completion of diagnostic and therapeutic orthopaedic procedures in children requires a high level of cooperation and pain control. This article provides a framework for safe and effective sedation of children undergoing such procedures. Following the definition of levels of sedation, the dynamics of the three phases of sedation (pre, intra, and post) are described. Specific sedative analgesic agents, monitoring, and transport are discussed. Finally, the art of working with children who require sedation is outlined. PMID- 11052052 TI - The cancer amputee and sexuality. AB - To the person experiencing amputation, the loss of a limb has a serious psychosexual impact. Whatever the age, the surgical procedure itself, pain, deformity, inability to perform simple customary acts, economic threat, and many other problems impose on the person facing the loss of their limb. Commonly, concentration is focused on functional abilities during and after prosthetic rehabilitation, and care taken to preserve the person's sexuality is often omitted. Major limb amputation can cause a decrease in self-esteem and body image due to perceived mutation, and this in turn can create emotional hurdles for both the patient and partner. Several potential problems such as the mechanics of body positioning during sex play, balance and movement, and phantom pain sensations can alter sexual function. Added difficulties include chemotherapy-induced neuropathies and enforced isolation because of limited mobility. Although these various disabling maladies may require alterations in a sexual relationship, sex drive and desire usually remain intact. Specific assessment techniques and interventions must be made available to enable the nurse to discuss sexual concerns at all stages of cancer and its treatment. As well, we must not fail to see alternatives to stereotypical behavior and acknowledge each patient's unique sexual identify. PMID- 11052053 TI - Venous stasis wounds. AB - Lower extremity wounds related to venous insufficiency are increasing. As the number of adults over the age of 65 increases, so do wounds related to changes in the vascular system. Venous stasis wounds can present as a real challenge to experienced practitioners. This article describes the many factors associated with venous wounds as well as the need for a comprehensive treatment plan. PMID- 11052054 TI - Total joint arthroplasty: a comparison of postacute settings on patient functional outcomes. AB - PURPOSE: This descriptive comparative study was conducted to examine functional outcomes of total joint arthroplasty patients discharged to subacute rehabilitation programs compared with those of patients discharged directly home with home physical therapy follow-up. SAMPLE: This study used a convenience sample of 96 patients having total joint arthroplasty performed by one physician within one institution. The postacute care setting was self-selected by the patients after information was provided on both options. METHODS: A patient self evaluation questionnaire was administered preoperatively and at 1 month and 3 months after surgery. A total functional score was obtained by combining the four subscores of the questionnaire/tool: subjective, pain, walking, and activities of daily living. Quantitative data on length of stay and cost were collected for each rehabilitative site. FINDING: All patients improved significantly over time in all subscores and in total score. There was no statistically significant difference in scores between the home and the subacute group. There was a significant difference in the mean total cost of the joint replacement for the subjects who went to the subacute unit ($24,144) compared to those who went directly home ($16,918). The groups were significantly different demographically, with the subacute group being older (age > 72), and likely to have comorbidities and to live alone. CONCLUSION: The majority of total joint replacement patients can achieve acceptable functional outcomes in a reasonable length of time at home with physical therapy supervision. Rehabilitation in a subacute facility may be most useful for the elderly patient with comorbidities, particularly those who live alone. PMID- 11052055 TI - Meeting the challenge of medication reactions in the elderly. AB - The number of elderly clients in most health care settings is increasing in direct proportion to the rapid aging of our general population. To plan and deliver optimum health care for elderly clients, nurses need to recognize this population's unique, special needs. One important aspect of quality care for older adults is managing their medications to assure the maximum beneficial results from medication therapy while avoiding adverse complications. Changes of aging will be examined in this article for their potential pharmacokinetic and pharmacodynamic impact. Incorporating this knowledge into practice will prepare professional nurses to act as change agents and advocates for increased awareness and sensitivity to actual or potential medication problems among all members of the health care team. PMID- 11052056 TI - Orthopaedic nursing core competencies: adult acute care. PMID- 11052057 TI - Introducing ... Mary Heron. PMID- 11052058 TI - The birth of love. PMID- 11052059 TI - Routine ultrasound in early pregnancy. PMID- 11052060 TI - How to calculate an EDD. PMID- 11052061 TI - Changes to maternity leave and the introduction of parental leave. PMID- 11052062 TI - Artificial feeding and risk. PMID- 11052063 TI - Antenatal visits. PMID- 11052064 TI - Sudden infant death syndrome. PMID- 11052065 TI - Calculating the EDD. PMID- 11052066 TI - Striving towards intrapartum excellence. PMID- 11052067 TI - Midwives, research ... and the consumption of fine wine. PMID- 11052068 TI - Making it real.... PMID- 11052069 TI - Pleased to meet you. Soo Downe. PMID- 11052070 TI - Birth by candlelight. PMID- 11052071 TI - Total syntheses of the Securinega alkaloids (+)-14,15-dihydronorsecurinine, (-) norsecurinine, and phyllanthine. AB - A new strategy for enantiospecific construction of the Securinega alkaloids has been developed and applied in total syntheses of (+)-14,15-dihydronorsecurinine (8), (-)-norsecurinine (6), and phyllanthine (2). The B-ring and C7 absolute stereochemistry of these biologically active alkaloids originated from trans-4 hydroxy-L-proline (10), which was converted to ketonitrile 13 via a high-yielding eight-step sequence. Treatment of this ketonitrile with SmI2 afforded the 6 azabicyclo[3.2.1]octane B/C-ring system 14, which is a key advanced intermediate for all three synthetic targets. Annulation of the A-ring of (-)-norsecurinine (6) with the required C2 configuration via an N-acyliminium ion alkylation was accomplished using radical-based amide oxidation methodology developed in these laboratories as a key step, providing tricycle 33. Annulation of the D-ring onto alpha-hydroxyketone 33 with the Bestmann ylide 45 at 12 kbar gave (+)-14,15 dihydronorsecurinine (8). In the securinine series, the D-ring was incorporated using an intramolecular Wadsworth-Horner-Emmons olefination of phenylselenylated alpha-hydroxyketone 47. The C14,15 unsaturation was installed late in the synthesis by an oxidative elimination of the selenoxide derived from tetracyclic butenolide 50 to give (-)-norsecurinine (6). The A-ring of phyllanthine (2) was formed from hydroxyketone 14 using a stereoselective Yb(OTf)3-promoted hetero Diels-Alder reaction of the derived imine 34 with Danishefsky's diene, affording adduct 35. Conjugate reduction and stereoselective equatorial ketone reduction of vinylogous amide 35 provided tricyclic intermediate 36, which could then be elaborated in a few steps to stable hydroxyenone 53 via alpha-selenophenylenone intermediate 52. The D-ring was then constructed, again using an intramolecular Wadsworth-Horner-Emmons olefination reaction to give phyllanthine (2). PMID- 11052072 TI - Variable strategy toward carbasugars and relatives. 1. Stereocontrolled synthesis of pseudo-beta-D-gulopyranose, pseudo-beta-D-xylofuranose, (pseudo-beta-D gulopyranosyl)amine, and (pseudo-beta-D-xylofuranosyl)amine. AB - Four novel, chiral nonracemic carbasugars have been synthesized from 1,2-O isopropylidene-D-glyceraldehyde. Furan- and pyrrole-based 2-silyloxy dienes- mimics of the alpha,gamma-dianions of gamma-hydroxy- and gamma-aminobutanoic acid, respectively--nicely served to complete the syntheses of two all-oxygen compounds, pseudo-beta-D-gulopyranose and pseudo-beta-D-xylofuranose, and two "anomeric" amino derivatives, (pseudo-beta-D-gulopyranosyl)amine (1,2,4-tri-epi validamine) and (pseudo-beta-D-xylofuranosyl)amine. Two sequential, highly diastereoselective carbon-carbon bond-forming maneuvers, i.e., a vinylogous crossed aldol addition and an intramolecular aldolization, proved central to these constructions. The fact that readily available heterocyclic diene scaffolds can be employed in the assembly of a varied repertoire of carbasugars and analogues widens the prospects of dienoxy silane chemistry. PMID- 11052073 TI - Formal total synthesis of (+)-diepoxin sigma. AB - The highly oxygenated antifungal anticancer natural product (+/-)-diepoxin sigma was prepared in 10 steps and in 15% overall yield from O-methylnaphthazarin. Highlights of the synthetic work include an Ullmann coupling and a possibly biomimetic oxidative spirocyclization for the introduction of the naphthalene ketal as well as the use of a retro-Diels-Alder reaction to unmask the reactive enone moiety in the naphthoquinone bisepoxide ring system. A novel highly bulky chiral binaphthol ligand was developed for a boron-mediated Diels-Alder reaction that constitutes a formal asymmetric total synthesis of (+)-diepoxin sigma. PMID- 11052074 TI - Functionalized carbosilane dendritic species as soluble supports in organic synthesis. AB - A new methodology, which is compatible with the use of reactive organometallic reagents, has been developed for the use of carbosilane dendrimers as soluble supports in organic synthesis. Hydroxy-functionalized dendritic carbosilanes Si[CH2CH2CH2SiMe2(C6H4CH(R)OH)]4 (G0-OH, R = H or (S)-Me) and Si[CH2CH2CH2Si[CH2CH2CH2SiMe2(C6H4CH(R)OH)]3]4 (G1-OH, R = H or (S)-Me) were prepared and subsequently converted into the esters Si[CH2CH2CH2SiMe2(C6H4CH(R)OC(O)CH2Ph)]4 (R = H or (S)-Me) and Si[CH2CH2CH2Si[CH2CH2CH2SiMe2(C6H4CH(R)OC(O)CH2C6H4 R')]3]4 (R = H and R' = H or R = (S)-Me and R' = H or R = H and R' = Br). As an example the latter compound was functionalized under Suzuki conditions. The functionalized carboxylic acid was obtained in high yield after cleavage from the dendritic support. Moreover, the ester functionalized dendrimers were converted to the corresponding zinc enolates followed by a condensation reaction with an imine to a beta-lactam in excellent yield and purity. Furthermore, it was demonstrated that a small combinatorial library of beta-lactams could be prepared starting from a carbosilane dendrimer functionalized with different ester moieties. These results show that carbosilane dendrimers can be applied as soluble substrate carriers for the generation of low molecular weight organic molecules. In combination with nanofiltration techniques, separation and recycling of the dendrimers can be realized. PMID- 11052075 TI - Eight-membered-ring solid-state conformational interconversion via the atom-flip mechanism, a CPMAS 13C NMR and crystallographic stereochemical study. AB - Nefopam methohalide (chloride, bromide, and iodide) medium-ring quaternary ammonium salts of the non-narcotic analgesic tertiary amine drug give crystals belonging to the identical monoclinic P2(1)/c space group, and all of these pseudopolymorphs exhibit the same packing motif. A singular boat-boat (BB) more compact conformation is observed in the nefopam methochloride crystal. Larger halide anions (bromide and iodide) increase the void distance between the 2(1) screw axis related adjacent ammonium cations to accommodate void-size dependent equilibrium quantities of the twist-chair-chair (TCC) more extended conformation. The BB:TCC occupancy factors are 0.961(5):0.039(5) [193 K], 0.780(5):0.220(5) [293 K], and 0.755(6):0.245(6) [343 K] for the methobromide crystal, while values of 0.657(5):0.343(5) [193 K] and 0.592(7):0.408(7) [293 K] were measured for the methiodide. Above a minimum of ca. 2.53 A, the occupancy factors were found to be linearly correlated to the intermolecular (TCC)Me(eq)-H...H-Me(ax)(TCC) distance between abutting methyl group protons in 2(1)-screw axis related neighbors. Temperature-dependent occupancy factors for the two conformers are interpreted in terms of a medium ring atom-flip facile interconversion between the two low energy conformations in crystals containing the appropriate size intercation void. A BB/TCC atom-flip interconversion in the methochloride unit cell would result in van der Waals interactions due to an estimated 2.31 A close intermolecular (TCC)Me(eq)-H...H-Me(ax)(TCC) distance between adjacent 2(1)-screw symmetry ammonium cations. The 203 K low-temperature CPMAS 13C NMR spectrum of the methiodide salt showed two slow exchange limit (SEL) delta 57.91 [BB] and delta 63.10 [TCC] OCH2CH2N peaks. A variable low-temperature CPMAS NMR investigation of the solid methiodide showed complex dynamic behavior that cannot be interpreted solely on the basis of an atom-flip conformational interconversion. Local magnetic fields from the gem-dimethyl rapidly rotating proton magnetic dipoles provide a distance-dependent T1 relaxation mechanism for neighboring carbons in the solid-state. PMID- 11052076 TI - New substituent effects of the trimethylsilyl group: photochemistry of 3 trimethylsilyl-2,5-cyclohexadienones and preparation of 4 alkylidenecyclopentenones. AB - The 4-acetoxymethyl-4-alkyl-3-trimethylsilyl-2,5-cyclohexadien-1-ones 9a-g were prepared from methyl 2-trimethylsilylbenzoate by the Birch reduction-alkylation reaction. Type A photorearrangements of 9a-g were regiospecific to give mixtures of two diastereomers of the corresponding 5-trimethylsilylbicyclo[3.1.0]hex-3-en 2-ones 11a-g. These bicyclohexenones are uniquely photostable; the diastereomers do not photointerconvert nor do they undergo the type B photorearrangement. Bicyclohexenones 11a-g undergo acid-catalyzed protiodesilylative rearrangement to give the 4-alkylidene-2-cyclopenten-1-ones 25a-g. It was of interest to find that the 4-(3'-butenyl)-2,5-cyclohexadienone 9e photorearranged to the 5 trimethylsilylbicyclo[3.1.0]hex-3-en-2-one 11e rather than undergoing the intramolecular 2 + 2 photocycloaddition. Furthermore, the 4-acetoxymethyl-3 methoxy-4-methyl-5-trimethylsilyl-2,5-cyclohexadienone 30a did not show type A photobehavior at 366 and 300 nm, while the 4-(3'-butenyl) analogue 30b gave the intramolecular 2 + 2 cycloadduct 31b. The effects of the trimethylsilyl and methoxy substituents on the photochemical reactivity of 2,5-cyclohexadien-1-ones are discussed from the perspective of n --> p* vs pi --> p* character of the triplet states of the dienones. PMID- 11052078 TI - Di-tert-butyl dicarbonate and 4-(dimethylamino)pyridine revisited. Their reactions with amines and alcohols AB - The reaction of BOC2O in the presence and absence of DMAP was examined with some amines, alcohols, diols, amino alcohols, and aminothiols. Often, unusual products were observed depending on the ratio of reagents, reaction time, polarity of solvent, pKa of alcohols, or type of amine (primary or secondary). In reactions of aliphatic alcohols with BOC2O/DMAP, we isolated for the first time carbonic carbonic anhydride intermediates; this helps explain the formation of symmetrical carbonates in addition to the O-BOC products. In the case of secondary amines, we succeeded to isolate unstable carbamic-carbonic anhydride intermediates that in the presence of DMAP led to the final N-BOC product. The effect of N methylimidazole in place of DMAP was also examined. PMID- 11052077 TI - Total synthesis of (+)-blastmycinone, (-)-litsenolide C1, and related natural trisubstituted lactones via alkynyltungsten compounds. AB - A general method for total synthesis of natural trisubstituted gamma-lactones is developed on the basis of the chemistry of alkynyltungsten compounds. The key step in this approach involves the cycloalkenation of tungsten-eta1-(3R,4S)-pent 1-yne-3,4-diol with aldehydes to give tungsten-oxacarbenium salts, further leading to 3-alkylidene-4-hydroxy-5-methyl-gamma-lactones upon demetalation. This synthetic sequence proceeds well for alkynylaldehydes and the MOM derivative of tungsten-eta1-(3R,4S)-pent-1-yne-3,4-diol. The resulting butyrolactone products are transformed into natural trisubstituted butyrolactones including (+) blastmycinone, (+)-blastmycinolactol, (+)-antimycinone, NFX-2, and (+) isodihydromahubanolide A. By using the same approach based on (R)-ethyl lactate, the natural (-)-litsenolide C1 can be prepared in a few steps. PMID- 11052079 TI - NAD(P)+-NAD(P)H models. 90. Stereoselection controlled by electronic effect of a carbonyl group in oxidation of NAD(P)H analog. AB - 4-Monodeuterated NAD(P)H model compounds (1,4,6,7-tetrahydro-1,6,11-trimethyl-5 oxo-5H-benzo[c]pyrido[2,3-e]az epin; 11Me-MMPAH) have been oxidized with a series of p-benzoquinone and its derivatives in the presence of Mg2+. The models have an axial chirality with respect to the orientation of carbonyl dipole, the dihedral angle of which is larger than 55 degrees out of the plane of dihydropyridine ring. Without Mg2+, the anti- (with respect to the carbonyl dipole) hydrogen is 3 to 32 times more reactive than the corresponding syn-hydrogen, whereas, when Mg2+ is present in the system, the selectivity is shifted toward the syn-preferency. Mg2+ plays the role of a Lewis acid catalyst to control the stereochemistry at the same time as it catalyzes the reaction. PMID- 11052080 TI - Formation of 1,10-disubstituted benzo AB - The first 1,10-heterodisubstituted benzo[c]cinnoline derivative 1 was prepared from the trinitrobiphenyl 2. Investigation of the mechanism of ring closure in 2, 5, and 8 revealed a complex reduction-oxidation-cyclization sequence. The mechanism is discussed in light of the stereoelectronic demands of the substituent functionalities. Benzo[c]cinnoline derivative 1 [C15H15N3S, monoclinic, P2(1)/c: a = 7.4063(3) A, b = 10.3739(5) A, c = 16.7642(8) A, beta = 91.816(1) degrees, Z = 4] and its 5-N-oxide 7(N5) [C18H18N3OS, triclinic, Pi: a = 8.1510(7) A, b = 8.6106(7) A, c = 12.102(1) A, alpha = 86.262(1) degrees, beta = 83.364(1) degrees, gamma = 74.711(1) degrees, Z = 4] were structurally characterized and showed a significant helical distortion of the heterocyclic ring. Oxidation of 1 with NCS or triamine 12 with PhI(OAc)2 led to a new heterocyclic ring system, ylide 13. Both benzo[c]cinnoline 1 and ylide 13 were characterized spectroscopically and the absorption spectra were correlated with the results of ZINDO calculations. PMID- 11052081 TI - A convenient strategy for the synthesis of 4,5-bis(o-haloaryl)isoxazoles AB - A series of new 1,2-bis(o-haloaryl)ethanones is efficiently prepared and applied to the synthesis of 4,5-bis(o-haloaryl)isoxazoles. Isolation of intermediate hydroxyisoxazolines, which are structurally examined, provides a definitive proof for a heterocyclization mechanism based on an amine exchange process. The isolation and X-ray crystallographic studies of significant side products such as benzamides and triarylpropionitriles are also described. PMID- 11052082 TI - Ascosalipyrrolidinone A, an antimicrobial alkaloid, from the obligate marine fungus Ascochyta salicorniae. AB - From the green alga Ulva sp., the endophytic and obligate marine fungus Ascochyta salicorniae was isolated. A. salicorniae was mass cultivated and found to produce the unprecedented and structurally unusual tetramic acid containing metabolites ascosalipyrrolidinones A (1) and B (2). Additionally, the new natural product ascosalipyrone (3) and the known metabolites 4 and 5 were obtained. Ascosalipyrrolidinone A (1) has antiplasmodial activity toward Plasmodium falciparum strains K1 and NF 54, as well as showing antimicrobial activity and inhibiting tyrosine kinase p56lck. PMID- 11052083 TI - Synthesis of cis,cis,cis-tetrasubstituted cyclobutanes. Trapping of tetrahedral intermediates in intramolecular nucleophilic addition AB - The intramolecular [2 + 2] photocycloaddition of bismaleimides leads to cage diimides 2. Nucleophilic addition on these compounds (NaBH4, RLi, or MeONa) gives rise to various diazatetracyclic 4 and 11 or oxadiazapentacyclic 3 "bowl shaped" alcohols. X-ray analyses of 3c and 11a provide definite structural data concerning these two highly functionalized compounds. PMID- 11052084 TI - Synthesis of enantiomerically pure alpha-substituted propargylic amines by reaction of organoaluminum reagents with oxazolidines. AB - Various oxazolidines prepared in two steps from (R)-phenylglycinol react at 0 degrees C with dialkylalkynylalane-triethylamine complexes in the presence of trimethylaluminum in high yield and diastereoselectivity. Enantiomerically pure primary alpha-substituted propargylamines can be easily obtained in two steps after removal of ferrocenylmethyl protective group under smooth acidic conditions and oxidative cleavage of the chiral appendage. PMID- 11052085 TI - Influence of anionic and cationic reverse micelles on nucleophilic aromatic substitution reaction between 1-fluoro-2,4-dinitrobenzene and piperidine AB - The nucleophilic aromatic substitution (S(N)Ar) reaction between 1-fluoro-2,4 dinitrobenzene and piperidine (PIP) were studied in two different reverse micellar interfaces: benzene/sodium 1,4-bis(2-ethylhexyl) sulfosuccinate (AOT)/water and benzene/benzyl-n-hexadecyl dimethylammonium chloride (BHDC)/water reverse micellar media. The kinetic profiles of the reactions were investigated as a function of variables such as surfactant and amine concentration and the amount of water dispersed in the reverse micelles, W0 = [H2O]/[surfactant]. In the AOT system at W0 = 0, no micellar effect was observed and the reaction takes place almost entirely in the benzene pseudophase, at every AOT and PIP concentration. At W0 = 10, a slight increment of the reaction rate was observed at low [PIP] with AOT concentration, probably due to the increase of micropolarity of the medium. However, at [PIP] > or = 0.07 M the reaction rates are always higher in pure benzene than in the micellar medium because the catalytic effect of the amine predominates in the organic solvent. In the BHDC system the reaction is faster in the micellar medium than in the pure solvent. Increasing the BHDC concentration accelerates the overall reaction, and the saturation of the micellar interface is never reached. In addition, the reaction is not base-catalyzed in this micellar medium. Thus, despite the partition of the reactants in both pseudophases the reactions effectively take place at the interface of the aggregates. The kinetic behavior can be quantitatively explained taking into account the distribution of the substrate and the nucleophile between the bulk solvent and the micelle interface. The results were used to evaluate the amine distribution constant between the micellar pseudophase and organic solvent and the second-order rate coefficient of S(N)Ar reaction in the interface. A mechanism to rationalize the kinetic results in both interfaces is proposed. PMID- 11052086 TI - Biogenetic syntheses of kopsijasminilam and deoxykopsijasminilam. AB - The hexacyclic ketoester 7, derived from cyclization of racemic minovincine (6), was reduced to two C-19 epimeric alcohols 8 and 9. Stereoelectronically controlled fragmentations of corresponding O-sulfonyl derivatives provided, respectively, the hexacyclic enamine 14 and, after oxidation of the olefin 16, the pentacyclic lactam 17 with a brigehead double bond. Formation of a carbamate, introduction of a second double bond at C-16, and conjugate reductive hydroxylation at C-20, or hydrogenation, gave the title products. PMID- 11052087 TI - Synthesis and conformational studies of peptidomimetics containing furanoid sugar amino acids and a sugar diacid. AB - Furanoid sugar amino acids (1) were synthesized and used as dipeptide isosteres to induce interesting turn structures in small linear peptides. They belong to a new variety of designed hybrid structures that carry both amino and carboxyl groups on rigid furanose sugar rings. Four such molecules, 6-amino-2,5-anhydro-6 deoxy-D-gluconic acid (3, Gaa) and its mannonic (4, Maa), idonic (5, Iaa), and a 3,4-dideoxyidonic (6, ddIaa) congeners were synthesized. The synthesis followed a novel reaction path in which an intramolecular 5-exo S(N)2 opening of the hexose derived terminal aziridine ring in 2 by the gamma-benzyloxy oxygen with concomitant debenzylation occurred during pyridinium dichromate oxidation of the primary delta-hydroxyl group to carboxyl function, leading to the formation of furanoid sugar amino acid frameworks in a single step. Incorporation of these furanoid sugar amino acids into Leu-enkephalin replacing its Gly-Gly portion gave analogues 8-11. Detailed structural analysis of these molecules by circular dichroism (CD) and various NMR techniques in combination with constrained molecular dynamics (MD) simulations revealed that two of these analogues, 8a and 10a, have folded conformations composed of an unusual nine-membered pseudo beta turn-like structure with a strong intramolecular H-bond between LeuNH --> sugarC3 OH. This, in turn, brings the two aromatic rings of Tyr and Phe in close proximity, a prerequisite for biological activities of opioid peptides. The analgesic activities of 8a,b determined by mouse hot-plate and tail-clip methods were similar to that of Leu-enkephalin methyl ester. The syn disposition of the beta-hydroxycarboxyl motif on the sugar rings appears to be the driving force to nucleate the observed turn structures in some of these molecules (8 and 10). Repetition of the motif on both sides of a furanose ring resulted in a novel molecular design of sugar diacid, 2,5-anhydro-D-idaric acid (7, Idac). Bidirectional elongation of the diacid moieties of 7 with identical peptide strands led to the formation of a C2-symmetric reverse-turn mimetic 12 which displayed a very ordered structure consisting of identical intramolecular H-bonds at two ends between LeuNH --> sugar-OH, the same as in 8 and 10. PMID- 11052088 TI - A simple method for the synthesis of substituted benzylic ketones: homologation of aldehydes via the in situ generation of aryldiazomethanes from aromatic aldehydes AB - A general method for the homologation of aldehydes to benzylic ketones has been developed. Aryldiazomethanes were generated in situ in the presence of an aldehyde by simply heating the tosylhydrazones of aromatic aldehydes in the presence of a stoichiometric amount of base in polar protic solvents. The resulting polar protic solvent promoted homologation afforded benzylic ketones in moderate to excellent yields with a variety of aldehydes. Isolation of the tosylhydrazones was not necessary; they could be prepared in ethanol and carried through the sequence without isolation. This methodology allows easy access to a wide variety of substituted aryldiazomethanes that would be difficult, or even impossible, to prepare via conventional methods and circumvents the toxicity and stability problems associated with the isolation and/or handling solutions of aryldiazomethanes. PMID- 11052089 TI - Sulfoxide-controlled S(N)2' displacements between cyanocuprates and epoxy vinyl sulfoxides. AB - Two short and convergent routes have been devised for the preparation of enantiomerically pure acyclic epoxy vinyl sulfoxides. These substrates undergo highly regio- and stereoselective S(N)2' displacements with lithium cyanocuprates to give alpha'-alkylated, gamma-oxygenated Z alpha,beta-unsaturated sulfoxides in moderate to good yields and with good to excellent diastereoselectivities. The absolute configuration of the newly formed carbon-carbon bond is primarily controlled by the chiral sulfur atom, which in a nonreinforcing situation can override the intrinsic anti tendency of the vinyl oxirane moiety and forces the cuprate to undergo syn addition. The hydroxy vinyl sulfoxide functionality of the resulting adducts should allow for subsequent asymmetric transformations thus enhancing the synthetic usefulness of this methodology. PMID- 11052090 TI - The effect of meta versus para substitution on the efficiency of chemiexcitation in the chemically triggered electron-transfer-initiated decomposition of spiroadamantyl dioxetanes. AB - A similar viscosity dependence of the CIEEL efficiencies for the para- and meta substituted spiroadamantyl dioxetanes 1 has been observed, which implies that an electron-transfer mechanism operates through solvent-caged species for both regioisomers. The pronounced difference in the chemiexcitation yields for the meta- and para-substituted dioxetanes is rationalized in terms of the much larger (ca. 200-fold) rate constant for the electron back-transfer (BET) step to afford the excited meta CIEEL emitter. An in-depth kinetic analysis of the viscosity effect on the excited-state generation for the para versus meta regioisomers supports this conclusion. PMID- 11052091 TI - Fluorinated tetrathiafulvalenes with preserved electron-donor properties and segregated fluorous bilayer structures based on F...F nonbonded interactions AB - The synthesis of the fluorinated 4,5-(2,2'-difluoropropylenedithio)-1,3-dithiol-2 one heterocycle with DAST allows for the preparation of several symmetrical and unsymmetrical di- and tetrafluoro-substituted tetrathiafulvalenes with accessible oxidation potentials (0.6 < E1(1/2) < 0.85 V vs SCE) despite the presence of the electron-withdrawing CF2 groups. The flexibility of the fluorinated seven membered ring identified from room-temperature NMR data has been thoroughly investigated by temperature-variable 1H and 19F NMR experiments, allowing for the identification of two independent folding processes, whose coalescence temperatures (Tc) and activation energies (deltaGdouble dagger) were determined (Tc = -15 degrees C, deltaGdouble dagger = 50.2 kJ mol(-1) and Tc = 47 degrees C, deltaGdouble dagger = 51.1 kJ mo(-1)). The analysis of the X-ray crystal structures of three of those fluorinated TTF demonstrates the efficiency of the nonbonded fluorine exclusion interactions for the stabilization of layered structures with fluorous bilayers, together with S...S van der Waals interactions and C-H...F hydrogen bonds. PMID- 11052092 TI - Efficient synthesis of (S)-4-phthalimido-1,3,4,5-tetrahydro-8-(2,6 dichlorobenzyloxy)- 3-oxo-2H-2-benzazepin-2-acetic acid (PHt-Hba(2,6-Cl2-Bn)-Gly OH). AB - 4-Amino-2-benzazepin-3-ones have proven very useful for studying the biologically active conformations of peptides. The synthesis of Pht-Aba-Xaa-OH by reaction of the corresponding 1,3-oxazolidin-5-one with trifluoromethanesulfonic acid (TFMSA) has been reported in the literature. However, when this procedure was applied to the preparation of Pht-Hba(Bn)-Gly-OH 8, many byproducts were formed and the yield of the desired aminobenzazepinones 7 and 8 was very low. We report in this paper an efficient methodology for the synthesis of Pht-Hba(2,6-Cl2-Bn)-Gly-OH 17 starting from the commercially available tyrosine. In our procedure, the dipeptide Pht-Tyr(2,6-Cl2-Bn)-Gly-OH 15 is converted to the 1,3-oxazolidin-5-one 16 which then undergoes Friedel-Crafts cyclization in the presence of tin tetrachloride to afford the desired 4-phthalimido-1,3,4,5-tetrahydro-8-(2,6 dichlorobenzyloxy)-2-be nzazepin-3-one 17 in excellent yield. PMID- 11052093 TI - Synthesis of vinyl- and alkynylcyclopentanetetraols by SmI2/Pd(0)-promoted carbohydrate ring-contraction. AB - A variety of vinyl- or alkynyl-substituted polyhydroxylated cyclopentanes and cyclobutanes are prepared in enantiomerically pure form from appropriate carbohydrate precursors, in a direct one-step ring-contraction procedure promoted by SmI2 and catalytic Pd(O). This reaction is thought to proceed through intermediate ring-opened allyl- or allenylsamarium complexes that undergo ring closure by intramolecular carbonyl addition. A predominant trans relationship is found between vinyl (or alkynyl) and hydroxyl groups at the two newly created stereogenic centers, with good to excellent levels of stereoselectivity being observed in the formation of homopropargyl cyclopentanol products. Under appropriate conditions, preparatively useful yields are realized of stereoisomers not directly available using alternative methodology. PMID- 11052094 TI - Efficient oxidation of alcohols to carbonyl compounds with molecular oxygen catalyzed by N-hydroxyphthalimide combined with a Co species AB - Highly efficient catalytic oxidation of alcohols with molecular oxygen by N hydroxyphthalimide (NHPI) combined with a Co species was developed. The oxidation of 2-octanol in the presence of catalytic amounts of NHPI and Co(OAc)2 under atmospheric dioxygen in AcOEt at 70 degrees C gave 2-octanone in 93% yield. The oxidation was significantly enhanced by adding a small amount of benzoic acid to proceed smoothly even at room temperature. Primary alcohols were oxidized by NHPI in the absence of any metal catalyst to form the corresponding carboxylic acids in good yields. In the oxidation of terminal vic-diols such as 1,2-butanediol, carbon-carbon bond cleavage was induced to give one carbon less carboxylic acids such as propionic acid, while internal vic-diols were selectively oxidized to 1,2 diketones. PMID- 11052095 TI - Asymmetric allylboration and ring closing alkene metathesis: a novel strategy for the synthesis of glycosphingolipids. AB - A novel strategy for the synthesis of D,L-glucosylceramide 1, a member of the glycosphingolipid class of natural products is described. Reagent-controlled asymmetric Brown allylboration gave excellent stereochemical control in the construction of adjacent stereocenters in the sphingoid base portion of the molecule. The trans-configured double bond was obtained as a single geometrical isomer by use of silicon-tethered olefin metathesis employing the Schrock carbene [(CF3)2MeCO]2Mo(=CHCMe2Ph)(=NC6H3-2,6-i-Pr2++ +) and in situ PhLi-induced ring opening of the intermediate 5,6-dihydro-2H-1,2-oxasiline followed by protodesilylation with TBAF in DMSO. The synthesis was completed by long chain amide formation and global deprotection. PMID- 11052096 TI - Synthesis and properties of oligonucleotides having a phosphorus chiral center by incorporation of conformationally rigid 5'-cyclouridylic acid derivatives. AB - This paper describes the design and synthesis of a conformationally rigid dimer building block Umpc3Um as a chiral center at the phosphate group with the S/N junction where c3 refers to a propylene bridge linked between the uracil 5 position and 5'-phosphate group of pUm. The extensive H1 NMR analysis of Umpc3Um suggests that the 5'-upstream Um has predominantly a C2'-endo conformation and the pc3Um moiety exists almost exclusively in a C3'-endo conformation. The absolute configuration of the diastereomers Umpc3Um(fast) (8a) and Umpc3Um(slow) (8b) was determined by CD spectroscopy as well as computer simulations. The oligonucleotides U4[Umpc3Um(fast)]U4 (13a) and U4[Umpc3Um(slow)]U4 (13b) incorporating 8a and 8b were synthesized by use of the phosphoramidite building blocks 11a and 11b, respectively. The Tm experiments of the duplexes formed between these modified oligomers and the complementary oligomers imply that the modified oligomer 13a having Umpc3Um(fast) has the Sp configuration at the chiral phosphoryl group. PMID- 11052097 TI - Total synthesis and antitumor activity of 12,13-desoxyepothilone F: an unexpected solvolysis problem at C15, mediated by remote substitution at C21. AB - A new epothilone analogue, 12,13-desoxyepothilone F (dEpoF, 21-hydroxy-12,13 desoxyepothilone B, 21-hydroxyepothilone D), was synthesized and evaluated for antitumor potential. A convergent strategy employed for the semipractical synthesis of 12,13-desoxyepothilone B (dEpoB) has been utilized to yield an amount of dEpoF sufficient for relevant biological studies. The results from an in vitro assay reveal that this new analogue is highly active against various tumor cell lines with a potency comparable to that of dEpoB. In particular, the growth of resistant tumor cells is inhibited by dEpoF at concentrations where paclitaxel (Taxol) is basically ineffective. A preliminary assessment of its in vivo activity is also promising. The new analogue, containing an additional hydroxyl group at C21, exhibits advantages over other epothilones in terms of water solubility, and can serve as a readily functionalizable handle to produce other useful compounds for pertinent biological studies. PMID- 11052099 TI - Chiral acetals as stereoinductors: diastereoface selective alkylation of dihydrobenzoxazine-derived amide enolates AB - Novel dihydrobenzoxazine-derived acetals of type 3 have been developed for asymmetric C-alkylations of propionyl amide enolates. High stereoselectivities are obtained for amides 15 and 22 which are rationalized in terms of intramolecular metal chelate formation. PMID- 11052098 TI - Novel antihyperglycemic terpenoid-quinones from Pycnanthus angolensis. AB - Two new compounds, pycnanthuquinone A (1) and pycnanthuquinone B (2), were isolated from leaves and stems of the African plant, Pycnanthus angolensis (Welw.) Warb (Myristicaceae), by bioassay-guided fractionation of an ethanolic extract using a diabetic mouse model. Pycnanthuquinones A and B are the first representatives of a novel terpenoid-type quinone skeleton, and both compounds possess significant antihyperglycemic activity. PMID- 11052100 TI - Fluorinated building blocks. The discovery of a stable difluoroallenyl indium and the synthesis of gem-difluoroallenyl and -propargyl synthons in aqueous media AB - The synthesis of two highly functional fluorinated motifs, TIPS-C triple bond C CF2-, and CF2=C=C(TIPS)- is described. This approach is mediated by a room temperature stable ethereal solution of a difluoroallene indium intermediate. This intermediate may be used as "stock solution" in the reaction with aqueous HCHO, to yield CF2=C=C(TIPS)CH2OH, and in a reaction with a Schiff base to produce the corresponding beta,beta-difluorohomopropargylamine. The synthetic potential of CF2=C=C(TIPS)CH2-OH has been demonstrated by its conversion to a difluorodihydrofuran derivative by a facile and efficient 5-endo-trig cyclization. Homopropargylic gem-difluoro alcohols are synthesized by addition of indium to a mixture of an aldehyde and TIPS-C triple bond C-CF2Br in predominantly aqueous media. PMID- 11052101 TI - Comparative study of the diastereoselective addition of allenyl zinc reagents to alpha-alkoxy (or silyloxy) aldehydes and imines. A straightforward synthesis of amino alcohols from imines AB - The addition of allenylzinc bromides to alpha-chiral imines proceeds with very high diastereoselectivity. This result is in contrast with the addition to the corresponding aldehydes, leading to poor diastereoselectivity. The anti/anti adducts are explained by Felkin-Ahn and Gaudemar-Yamamoto models of the transition state. PMID- 11052102 TI - Unique regio- and stereoselectivity in Pd-catalyzed chlorocarbonylation reaction of 2-phenylethynyl selenides and 2-alkylethynyl selenides. Highly stereoselective synthesis of 2-seleno-3-chloroacrylates AB - Regio- and stereoselectivity in the chloropalladation carbonylation reaction of different acetylenic selenides in the presence of 0.05 equiv of PdCl2 and 3 equiv of cupric chloride under 1 atm of carbon monoxide affording 2-seleno-3 chloroacrylates were investigated. Opposite stereoselectivities were observed with 2-phenylethynyl selenides and 2-alkylethynyl selenides: the reactions of 2 phenylethynyl selenides afforded (E)-2-seleno-3-chloro-3-phenylacrylates, while the reactions of 2-alkylethynyl selenides gave (Z)-2-seleno-3-chloro-3 alkylacrylates. A chloropalladation carbonylation mechanism for this reaction was proposed. The regio- and stereoselective chloropalladation of the carbon-carbon triple bond in acetylenic selenides affords 1-enylpalladium intermediates, in which the palladium atom connects with the carbon atom bonding with the selenium atom. Carbonylation in the presence of an alcohol affords 2-seleno-3 chloroacrylates. PMID- 11052103 TI - Benzocyclobutadienyl anion: formation and energetics of an antiaromatic molecule AB - Benzocyclobutadienyl diazirine (2) was synthesized and reacted with hydroxide ion in a Fourier transform mass spectrometer to afford the conjugate base of benzocyclobutadiene (1a). Authentication of the ion structure was carried out by a derivatization experiment (i.e., la was converted to benzocyclobutenone enolate, which has previously been studied), and its reactivity was explored. Thermochemical data for benzocyclobutadiene (1) were obtained (deltaH(o)acid (1) = 386 +/- 3 kcal mol(-1), EA(1r) = 1.8 +/- 0.1 eV, and C-H BDE (1) = 114 +/- 4 kcal mol(-1)), compared to MP2 and B3LYP calculations, and contrasted to a series of model compounds. Cyclobutadienyl radical appears to be quite different from benzocyclobutadienyl radical (1r) and worth further exploration. PMID- 11052104 TI - Reactivity of 3-iodoimidazo[1,2-a]pyridines using a Suzuki-type cross-coupling reaction. AB - The influence of base and solvent in Suzuki cross-coupling reaction on various 2 substituted-3-iodoimidazo[1,2-a]pyridines was reported. The reactivity was largely influenced by nature of the substituent. Optimized yields and shortened times of reaction were obtained using strong bases in DME. PMID- 11052105 TI - Synthesis of functionalized cannabinoids. AB - An effective synthesis of tricyclic, nonclassical cannabinoids has been developed on the basis of a cation-olefin cyclization that forms the two nonaromatic rings with the desired stereochemistry in a single step. PMID- 11052106 TI - Ester hydrolysis and enol nitrosation reactions of ethyl cyclohexanone-2 carboxylate inhibited by beta-cyclodextrin AB - Both the ester hydrolysis and the nitrosation reactions of the enol tautomer of ethyl cyclohexanone-2-carboxylate (ECHC) are investigated in the absence and presence of beta-cyclodextrin (beta-CD). The ester hydrolysis reaction is studied in dilute H2O and D2O solutions of hydrochloric acid and in aqueous buffered solutions of carboxylic acids (acetic acid and its chloro derivatives). The pseudo-first-order rate constant increases with both the [H+] and the total buffer concentration, indicating that the hydrolysis is subject to acid and general base catalysis. Substantial solvent isotope effects in the normal direction (kH/kD > 1) for the acid-catalyzed hydrolysis was observed. Addition of beta-CD strongly slows the hydrolysis reaction. The variation of the observed rate constant (k(o)) with [beta-CD] exhibits saturation behavior, consistent with 1:1 binding between the enol of ECHC and beta-CD. The binding is quite strong, and bound ECHC-enol is unreactive. The nitrosation reaction of ECHC in aqueous acid medium, using sodium nitrite in great excess over the concentration of ECHC, yields perfect first-order kinetics, indicating that the slow step is the nitrosation of the enol tautomer. This finding suggests that a great percentage of the total ECHC concentration must exist in the enol form. The nitrosation reaction is of first order in [nitrite] and is catalyzed by the presence of Cl-, Br-, or SCN- ions, which indicates that the attack of the nitrosating agent is the slow step. The nitrosation reaction is also strongly inhibited by the presence of beta-CD because of the formation of unreactive inclusion complexes between the host, beta-CD, and the guest, the enol of ECHC. In alkaline medium, the formation of the enolate ion is observed, which absorbs at higher wavelengths (lambda(max) = 256 nm in acid medium shifts to lambda(max) = 288 nm in alkaline medium). This anion also undergoes ester hydrolysis spontaneously, but shows neither specific basic catalysis nor appreciable effect by the presence of beta CD. From kinetic and spectroscopic measurements the pKa of the enol of ECHC has been determined as 12.35. PMID- 11052107 TI - (R)-tert-Butoxycarbonylamino-fluorenylmethoxycarbonyl-glycine from (S) benzyloxycarbonyl-serine or from papain resolution of the corresponding amide or methyl ester. AB - The enantiospecific synthesis of (R)-Boc-(Fmoc)-aminoglycine 7 was achieved. (S) Cbz-serine 1 was reacted with diphenylphosphoryl azide in the presence of triethylamine to yield cyclic (S) carbamate 2. The ring nitrogen of 2 was protected with a Boc group (3). The cyclic carbamate of 3 was hydrolyzed with benzyltrimethylammonium hydroxide to yield the (R)-enantiomer of alcohol 4. The oxidation of 4 with pyridinium dichromate yielded the enantiomerically pure (97% ee) (R)-Boc-(Cbz)-aminoglycine 5, which was converted to 7 with retention of optical purity. Similarly, starting from (S)-Boc-serine 9, cyclic (S) carbamate 10 was obtained. The ring nitrogen of 10 was protected with a Cbz group (11) with retention of configuration. The cyclic carbamate of 11 was base hydrolyzed to yield 12, the (S)-enantiomer alcohol. Independently, Boc-(Fmoc)-aminoglycine amide 13 and Boc-(Fmoc)-aminoglycine methyl ester 14 were resolved using papain. The stereochemistry of the isolated acid was determined to be (R) by coelution on HPLC of its derivative with Marfey's reagent and that of an authentic sample (7) obtained by enantiospecific synthesis. PMID- 11052108 TI - On the behavior of alpha,beta-unsaturated thioaldehydes and thioketones in the Diels-Alder reaction. AB - Alpha,beta-unsaturated thioaldehydes and thioketones, R1CH=CH-C(=S)R2, are prepared in situ by the reaction between the corresponding carbonyl compounds and bis(dimethylaluminum) sulfide. These compounds undergo [4 + 2] self-dimerization reactions in which one molecule serves as the heterodiene component and the other as the dienophile to afford different types of dimeric products depending on the R1 and R2: 1,2-dithiin and 1,3-dithiin (R1 = R2 = H), 1,2-dithiin (R1 = Ph, R2 = H, CH3), or dihydrothiopyran (R1 = R2 = Ph). These differences in selectivity are explained on the basis of the relative energies evaluated by molecular orbital (MO) calculations at the DFT (density functional theory) level. The calculations show that in the dimerization reaction of thioacrolein (I), the head-to-tail (S-C S bonded) dimers are kinetically more stable by about 5 kcal/mol but slightly thermodynamically unstable by about 2 kcal/mol than the head-to-head (S-S bonded) dimers. The calculations on thiocinnamaldehyde (IV) indicate that the dimerization reactions of phenyl-substituted alpha,beta-unsaturated thioaldehydes and thioketones are almost equally controlled by thermodynamic and kinetic factors. These unsaturated thiocarbonyl compounds also function as heterodienes (C=C-C=S) in the cycloaddition reaction with norbornadiene and as dienophiles (C=S) in the reaction with cyclopentadiene. PMID- 11052109 TI - Computational studies on the BF3-catalyzed cycloaddition of furan with methyl vinyl ketone: a new look at Lewis acid catalysis AB - Transition structures for both uncatalyzed and BF3-catalyzed Diels-Alder reactions involving furan and methyl vinyl ketone have been determined at the hybrid DFT (B3LYP/6-31G*) level of theory. The transition structures are predicted to be relatively concerted and highly asynchronous in all cases. A subsequent bond-order analysis has been carried out at the MP2/6-31G*//B3LYP/6 31G*. The role of the Lewis acid and the origin of the endo selectivity have been discussed in terms of the nature and number of interactions present in the four possible transition structures. The partition of the potential energy barrier has also been used to estimate the contributions of the pure deformation energy and the differential interaction between the reaction partners on passing from the ground state to the saddle point. This analysis reveals that the major influence arises from the heterodiene-dienophile interaction instead of that corresponding to a BF3-dienophile interaction. PMID- 11052110 TI - Ab initio studies of benzocyclopropenone, benzocyclopropenone-containing AB - Ab initio calculations were carried out on cyclopropenone, 1, benzocyclopropenone, 2, the benzocyclopropenone-containing [2.2]paracyclophane derivative tetracyclo[8.3.2.(4,7)O(11,13)]heptadeca-1(13),4,6,10,14,16-hexaen-12 one, 3, its decarbonylation product tricyclo[8.2.2.2(4,7)]hexadeca-1(12), 4,6,10,13-pentaen-15-yne, 5, a benzyne intermediate, and the bridged benzobarrelene derivative, pentacyclo[5.5.2.2.(1,4)O(4,14)O(10,13)]hexadeca 2,7,9,13,15-pentaene, 6. These calculations suggest that benzocyclopropenone containing [2.2]paracyclophane, 3, and highly strained bridged benzobarrelene, 6, could exist as stable species. Both aryl rings of the benzocyclopropenone derivative 3 are predicted to be distorted from planarity. This distortion relieves some angle strain present in planar benzocyclopropenone due to the presence of the annulated three-membered ring. Calculations on benzobarrelene, 8, and [2.2]paracyclophane, 4, were performed for comparison to gain a better understanding of the strain borne in bridged benzobarrelene 6. The activation barrier for the intramolecular [4 + 2] cycloaddition of 5 to give 6 was estimated at 18.8 kcal/mol while that for the corresponding [2 + 2] cycloaddition, giving the less stable 9, was 54.5 kcal/mol. The [2 + 2] cycloaddition's transition state was twisted in a manner reminiscent of the conservation of orbital symmetry prediction for an unstrained system. PMID- 11052111 TI - Intramolecular cyclization of beta-amino and beta-ammonio radicals: a new synthetic route to the 1-azabicyclo AB - Treatment of 1-(2-phenylselenoethyl)-1,2,5,6-tetrahydropyridine (15) with tributyltin hydride affords only the product of reduction, demonstrating the reluctance of the 5-hexenyl radical 9 to undergo ring closure. When the nature of the radical is modified, either by introduction of an ester group at C4 or via its quaternary ammonium salt, cyclization occurs readily; while the radical 52 gives an excellent yield of 1-methyl-1-azoniabicyclo[3.2.1.]octyl bromide (55) uncontaminated with the product of reduction, the bicyclic product from 21 is accompanied by some reduced material. Production of the unwanted alkene can be eliminated in the latter by recourse to the quaternary ammonium ester 1-(2 bromoethyl)-4-carbethoxy-1-methyl-1,2,5,6-tetrahydropyridinium bromide (35) which, when exposed to tributyltin hydride, affords a 1:1 endo/exo mixture of 4 carbethoxy-1-methyl-1-azoniabicyclo[3.2.1]octyl bromide (37) exclusively. These results support the demonstration of the powerful polar effect of an ester function when attached to the double bond of a 5-hexenyl system, a property which can be exploited in the case of the radical 58. Treatment of the precursor, 1-(2 bromoethyl)-3-carbethoxy-1-methyl-3-pyrrolinium bromide (60), with tributyltin hydride generates 58 which is found to cyclize with high regioselectivity, affording a convenient high-yielding synthesis of the endo/exo isomers of 3 carbethoxy-1-methyl-1-azoniabicyclo[2.2.1]heptyl bromide 57. The isomeric bicyclo[2.2.1]heptyl ester 63 was not detected. These observations are in accordance with predictions based upon frontier molecular orbital considerations. PMID- 11052112 TI - A tightly coupled linear array of perylene, bis(porphyrin), and phthalocyanine units that functions as a photoinduced energy-transfer cascade AB - We have prepared a linear array of chromophores consisting of a perylene input unit, a bis(free base porphyrin) transmission unit, and a free base phthalocyanine output unit for studies in artificial photosynthesis and molecular photonics. The synthesis involved four stages: (1) a rational synthesis of trans AB2C-porphyrin building blocks each bearing one meso-unsubstituted position, (2) oxidative, meso,meso coupling of the zinc porphyrin monomers to afford a bis(zinc porphyrin) bearing one phthalonitrile group and one iodophenyl group, (3) preparation of a bis(porphyrin)-phthalocyanine array via a mixed cyclization involving the bis(free base porphyrin) and 4-tert-butylphthalonitrile, and (4) Pd mediated coupling of an ethynylperylene to afford a perylene-bis(porphyrin) phthalocyanine linear array. The perylene-bis(porphyrin)-phthalocyanine array absorbs strongly across the visible spectrum. Excitation at 490 nm, where the perylene absorbs preferentially, results in fluorescence almost exclusively from the phthalocyanine (phi(f) = 0.78). The excited phthalocyanine forms with time constants of 2 ps (90%) and 13 ps (10%). The observed time constants resemble those of corresponding phenylethyne-linked dyads, including a perylene-porphyrin (< or = 0.5 ps) and a porphyrin-phthalocyanine (1.1 ps (70%) and 8 ps (30%)). The perylene-bis(porphyrin)-phthalocyanine architecture exhibits efficient light harvesting properties and rapid funneling of energy in a cascade from perylene to bis(porphyrin) to phthalocyanine. PMID- 11052114 TI - Pauson-Khand reaction of optically active 6,7-bis(tert-butyldimethylsiloxy)non-1 en-8-ynes AB - Treatment of (6S,7S)-7-bis(tert-butyldimethylsiloxy)non-1-en-8-ynes with dicobalt octacarbonyl gave the corresponding cobalt complex. This complex was subsequently exposed to the Pauson-Khand conditions in the presence of a promoter such as cyclohexylamine, thioanisole, methyl isopropyl sulfide, and butyl methyl sulfide ending up with the stereoselective production of the (2S,3S,6S,7S)-7 methylbicyclo[4.3.0]nonenone derivatives instead of the expected (2S,3S,7S) bicyclo[5.3.0]decenone species. PMID- 11052113 TI - Asymmetric homologation of boronic esters bearing azido and silyloxy substituents. AB - In the asymmetric homologation of boronic esters with a (dihalomethyl)lithium, substituents that can bind metal cations tend to interfere. Accordingly, we undertook the introduction of weakly basic oxygen and nitrogen substituents into boronic esters in order to maximize the efficiency of multistep syntheses utilizing this chemistry. Silyloxy boronic esters cannot be made efficiently by direct substitution, but a (hydroxymethyl)boronic ester has been silylated in the usual manner. Conversion of alpha-halo boronic esters to alpha-azido boronic esters has been carried out with sodium azide and a tetrabutylammonium salt as phase-transfer catalyst in a two-phase system with water and either nitromethane or ethyl acetate. These are safer solvents than the previously used dichloromethane, which can form an explosive byproduct with azide ion. Boronic esters containing silyloxy or alkoxy and azido substituents have been shown to react efficiently with (dihalomethyl)lithiums, resulting in efficient asymmetric insertion of the halomethyl group into the carbon-boron bond. PMID- 11052115 TI - Total synthesis of oxidized phospholipids. 3. The (11E)-9-hydroxy-13-oxotridec-11 enoate ester of 2-lysophosphatidylcholine. AB - A total synthesis of (11E)-9-hydroxy-13-oxotridec-11-enoate ester of 2 lysophosphatidylcholine (HOT-PC) was devised to facilitate identification of this oxidized phospholipid. A lactone, 8-(3-oxo-1H,6H-2-oxinyl)octanoic acid (1), believed to be generated through an intermediate (11E)-9-hydroxy-13-oxotridec-11 enoic acid (HOT), is produced upon autoxidation of linoleic acid. A synthesis of lactone 1 methyl ester was accomplished from HOT involving a novel trans-cis isomerization that is driven to completion by cyclization to a hemiacetal. An alternative route to this carbon skeleton was also achieved that provides the lactone 1 itself. PMID- 11052116 TI - An efficient synthesis of a new series of acyclonucleosides starting from beta amino alcohols. AB - A series of new acyclonucleosides analogues 3 has been synthesized very efficiently in three steps starting from beta-amino alcohols 1. The key step of this process is a nucleophilic substitution with various nucleophiles on 2,2' anhydronucleosides 2. The chemo- and stereoselectivities of this reaction are discussed. AM1 calculations sustained the observed chemoselectivity. PMID- 11052117 TI - Synthesis and applications of [1-(15)N]-labeled 4,6-dimethyl-4H [1,2,5]oxadiazolo[3,4-d]pyrimidine-5,7-dione 1-oxide as a useful tool for mechanistic investigations. AB - [1-(15)N]-Labeled 4,6-dimethyl-4H-[1,2,5]oxadiazolo[3,4-d]pyrimidine-5,7-dione 1 oxide (1-(15)N1) was easily prepared by nitration of commercially available 6 amino-1,3-dimethyl-1H-pyrimidine-2,4-dione using 15N-enriched nitric acid followed by an intramolecular oxidative cyclization with iodosylbenzene diacetate under mild conditions. On the basis of the experimental results using 1-(15)N1, the formation of 8-phenyltheophylline (3), the 1,3-dimethylalloxazines (4: n = 0, 1), and 1,3,7,9-tetramethyl-1H,9H-pyrimido[5,4-g]pteridine-2,4,6,8-tetraone++ + (5) in the thermal reaction of the N-oxide 1 with benzylamine, aniline, or piperidine, and the generation of NO or NO-related species in the reaction with N acetylcysteamine were reasonably explained by considering the initial attack of the employed nucleophiles on the 3a-position of 1. PMID- 11052118 TI - A [4 + 4] 2-pyridone approach to taxol. 3. Stereocontrol during elaboration of the cyclooctane. AB - Intramolecular photocycloaddition of 2-pyridones connected through a four-carbon tether (6-[4-(1,2-dihydro-1-methyl-2-oxo-3-pyridinyl)-4-[[(1,1-dimethylethyl)++ +dimethylsilyl]oxy]butyl]-4-methoxy-1,3-dimethyl-2(1H)-pyridinone) yields a single tetracyclic product with four new stereogenic centers. The diastereoselectivity of this [4 + 4] reaction is fully controlled by a stereogenic carbon of the tether. Treatment of the photoproduct with osmium tetraoxide transforms the alkene to a diol and the enol ether to an alpha-hydroxy ketone, with stereocontrol dictated by nearby lactams that block one face of each alkene. Allylmagnesium bromide addition to the ketone also yields a single diastereomer, but unexpectedly this product results from approach of the nucleophile to the most-hindered face of the ketone. Study of this reaction in a model system has found the allylic nucleophile to be unique, with nonallylic reagents approaching along the expected, least-hindered path. This contrasteric addition likely results from coordination of the allylic nucleophile to the nearby amide. The amide can therefore act either as a steric shield or as a directing group. The three steps of photocycloaddition, cis-hydroxylation, and nucleophilic addition constructs both quaternary carbons of the cyclooctane and four of the five stereogenic centers found in the eight-membered ring of Taxol. PMID- 11052119 TI - The cumbiasins, structurally novel diterpenes possessing intricate carbocyclic skeletons from the West Indian sea whip Pseudopterogorgia elisabethae (Bayer). AB - From the hexane extract of the West Indian gorgonian Pseudopterogorgia elisabethae, two diterpenes, cumbiasins A (1) and B (2), having a novel tetracyclic carbon skeleton named cumbiane, have been isolated. In addition, we have isolated cumbiasin C (3), a ring cleavage product of cumbiasin B that possesses an unusual carbocyclic framework named seco-cumbiane. The structures and relative configurations of metabolites 1-3 were elucidated by interpretation of overall spectral data, which included 2D NMR correlation methods, IR, UV, and accurate mass measurements (HREI-MS and HRFAB-MS). The carbocyclic skeletons of the cumbiasins are unprecedented and represent new classes of C20 rearranged diterpenes. Cumbiasins A and B display mild in vitro anti-tuberculosis activity. PMID- 11052120 TI - Rational design of highly diastereoselective, organic base-catalyzed, room temperature Michael addition reactions. AB - Via the rational design of a single-preferred transition state, stabilized by electron donor-acceptor-type attractive interactions, structural and geometric requirements for the corresponding starting compounds have been determined. The Ni(II) complex of the Schiff base of glycine with o-[N-alpha picolylamino]acetophenone, as a nucleophilic glycine equivalent, and N-(trans enoyl)oxazolidin-2-ones, as derivatives of an alpha,beta-unsaturated carboxylic acid, were found to be the substrates of choice featuring geometric/conformational homogeneity and high reactivity. The corresponding Michael addition reactions were found to proceed at room temperature in the presence of catalytic amounts of DBU to afford quantitatively the addition products with virtually complete diastereoselectivity. Acidic decomposition of the products followed by treatment of the reaction mixture with NH4OH gave rise to the diastereomerically pure 3-substituted pyroglutamic acids. PMID- 11052121 TI - Molecular addition compounds. 17. Borane and chloroborane adducts with organic sulfides for hydroboration AB - The following sulfides have been examined as borane carriers in comparison with dimethyl sulfide and 1,4-oxathiane: tert-butyl methyl sulfide, isoamyl methyl sulfide, ethyl isoamyl sulfide, tert-butyl isoamyl sulfide, diisoamyl sulfide, tetrahydrothiophene, tetrahydro-thiopyran, thioanisole, 3 ethylthiotetrahydrofuran, bis(3-tetrahydrofuryl) sulfide, and bis(2-methoxyethyl) sulfide. Their complexing ability toward borane increases in the following order: thioanisole < ether-sulfides < dialkyl sulfides < dimethyl sulfide. Borane adducts of the sulfides are liquids above 0 degrees C. The thioanisole adduct loses diborane at room temperature. The reactivity of the adducts toward 1-octene increases in the reversed order of the complexing ability of the sulfides. Diisoamyl sulfide has a mild, ethereal, agreeable aroma, its synthesis is economical and the borane adduct, 4.2 M in BH3, is stable over prolonged periods at room temperature. The sulfide can be recovered from hydroboration-oxidation products by distillation. Consequently, diisoamyl sulfide is a new promising borane carrier. Bis(2-methoxyethyl) sulfide, easily synthesized from the low cost thiodiethanol, is three times more soluble in water than 1,4-oxathiane. Its borane adduct is 6.0 M in BH3 and can substitute for more expensive borane-1,4 oxathiane in hydroboration reactions. Applications of these new borane adducts in the synthesis of mono- and dichloroborane adducts was also studied. The equilibrium ratios observed for the new chloroborane adducts were similar to that observed for dimethyl sulfide adducts. However, the hydroboration of 1-octene with these new chloroborane adducts are much faster than the corresponding adducts of dimethyl sulfide, which are currently used extensively. PMID- 11052122 TI - Stereoselective synthesis of 7,11-guaien-8,12-olides from santonin. Synthesis of podoandin and (+)-zedolactone A. AB - Photochemical rearrangement of hydroxy ester 2, easily obtained from santonin (1), afforded butenolide 4, a good starting material for the synthesis of 7,11 guaien-8,12-olides. Compound 4 has been transformed into compound 10, which has been used for the synthesis of podoandin (5) and (+)-zedolactone A (ent-6). Regioselective elimination of the acetyl group on C10 afforded directly podoandin (5). For the synthesis of ent-6, a hydroxyl group has been regio- and stereoselectively introduced at the 4alpha-position through the 3alpha,4alpha epoxide 15. The basic hydrolysis of the 10-acetyl group in compound 18 took place with concomitant intramolecular conjugated addition of the alkoxide to the butenolide moiety to give ether 19. Cleavage of the 7,10-oxido bridge via the lactone enolate afforded (+)-zedolactone A (ent-6). This synthesis has allowed for the establishment of the absolute stereochemistry of natural zedolactone A as the enantiomer of our synthetic product. PMID- 11052123 TI - Substituent effects in pericyclic reactions of radical cations: the ring opening of 3-substituted cyclobutene radical cations AB - The substituent effects on the ring-opening reaction of cyclobutene radical cations have been studied at the Becke3LYP/6-31G* level of theory. The effect on the reaction energies and activation energies of the concerted and stepwise pathways of electron-donating substituents such as methyl and methoxy as well as electron-withdrawing substituents such as nitrile and carboxaldehyde in the 3 position of the cyclobutene is discussed. The exothermicity of the reaction correlates well with the ability of the substituent to stabilize the 1,3 butadiene radical cation by electron donation or conjugation. The relative stability of the (E) and (Z) isomers of the resulting 1,3-butadiene radical cations depends largely on steric effects. Similarly, steric effects are responsible for the relative energies of the different diastereomeric transition structures. The cyclopropyl carbinyl intermediate of the stepwise pathway resembles the nonclassical carbocation and is stabilized by electron-donating substituents. In the case of electron-donating substituents, this species becomes a minimum on the potential energy hypersurface, whereas unstabilized or destabilized cyclopropyl carbinyl radical cations are not minima on the hypersurface. The stabilization of the cyclopropyl carbinyl radical cation by substituents correlates qualitatively with the Brown-Okamoto substituent parameter sigma+. However, in all cases studied here, the concerted mechanism is the lowest energy pathway. PMID- 11052124 TI - A computational study of the hydroxy-group directivity in the peroxyformic acid epoxidation of the chiral allylic alcohol (Z)-3-methyl-3-penten-2-ol: control of threo diastereoselectivity through allylic strain and hydrogen bonding AB - Eight transition structures for the epoxidation of the chiral allylic alcohol (Z) 3-methyl-3-penten-2-ol (1) with peroxyformic acid have been computed by the B3LYP density functional method with 6-31G(d) and 6-31G(d,p) basis sets. The four lowest-energy transition structures and their respective pre-reaction clusters were fully re-optimized by employing 6-311+G(d,p) and correlation-consistent polarized valence triple-zeta cc-pZTV basis sets. The relative energies of the transition structures were found to be highly sensitive to the basis set applied. The transition state for threo product formation, anti-(2S,3R,4S)-TS-3f, with the lowest total energy (at B3LYP/611+G(d,p) and B3LYP/AUG-cc-pZTV) of all the TSs examined, has a planar peracid moiety and is a precursor for the 1,4 migration of the peracid hydrogen atom Ha to the peroxy oxygen atom O4. The use of different basis sets does not affect markedly the geometry of the anti-(2S,3R,4S)-TS-3f transition structure. The transition state for erythro epoxidation, syn (2R,3R,4S)-TS-3a, is 0.9 kcal/mol higher in energy and possesses a nonplanar peracid approaching the C=C bond in a manner intermediate between spiro and planar. The relative energy and nonplanarity of this syn transition structure is highly sensitive to the basis set applied. With the smaller basis set, 6 31G(d,p), it is actually the lowest-energy TS and the peracid moiety is significantly skewed. The contribution of the four lowest energy transition stuctures 3a, 3b, 3e, and 3f to the threo/erythro product ratio has been assessed through an extended Curtin-Hammet principle analysis of this multi-transition state reaction. It has been found that this approach agrees well with the experimental threo/erythro product ratio, in particular when the corrections for a solvent effect are made within the self-consistent isodensity polarized continuum model (SCI-PCM). PMID- 11052125 TI - N-NO bond dissociation energies of N-nitroso diphenylamine derivatives (or analogues) and their radical anions: implications for the effect of reductive electron transfer on N-NO bond activation and for the mechanisms of NO transfer to nitranions AB - The heterolytic and homolytic N-NO bond dissociation energies [i.e., deltaHhet(N NO) and deltaHhomo(N-NO)] of 12 N-nitroso-diphenylamine derivatives (1-12) and two N-nitrosoindoles (13 and 14) in acetonitrile were determined by titration calorimetry and from a thermodynamic cycle, respectively. Comparison of these two sets of data indicates that homolysis of the N-NO bonds to generate NO* and nitrogen radical is energetically much more favorable (by 23.3-44.8 kcal/mol) than the corresponding heterolysis to generate a pair of ions, giving hints for the driving force and possible mechanism of NO-initiated chemical and biological transformations. The first (N-NO)-* bond dissociation energies [i.e., deltaH(N NO)-* and deltaH'(N-NO)-*] of radical anions 1-*-14-* were also derived on the basis of appropriate cycles utilizing the experimentally measured deltaHhet(N-NO) and electrochemical data. Comparisons of these two quantities with those of the neutral N-NO bonds indicate a remarkable bond activation upon a possible one electron transfer to the N-NO bonds, with an average bond-weakening effect of 48.8 +/- 0.3 kcal/mol for heterolysis and 22.3 +/- 0.3 kcal/mol for homolysis, respectively. The good to excellent linear correlations among the energetics of the related heterolytic processes [deltaHhet(N-NO), deltaH(N-NO)-*, and pKa(N-H)] and the related homolytic processes [deltaHhomo(N-NO), deltaH'(N-NO)-*, and BDE(N H)] imply that the governing structural factors for these bond scissions are similar. Examples illustrating the use of such bond energetic data jointly with relevant redox potentials for analyzing various mechanistic possibilities for nitrosation of nitranions are presented. PMID- 11052127 TI - Synthesis and characterization of 5,10-bis(2-thienyl)indeno PMID- 11052126 TI - Rapid access to enantiopure bicyclic diamines via aza-Diels-Alder reaction of iminoamides PMID- 11052128 TI - Synthesis of a peptidomimetic tricyclic tetrahydrobenzo[ij]quinoline as a VLA-4 antagonist. PMID- 11052129 TI - Facile ring opening of oxiranes with aromatic amines in fluoro alcohols PMID- 11052130 TI - A new approach for the chemoselective debromination of chiral bromohydrins. Toward the development of a very general approach to enantiopure alpha unsubstituted beta-hydroxy acids PMID- 11052131 TI - Catalytic asymmetric oxidation of sulfides to sulfoxides mediated by chiral 3 substituted-1,2-benzisothiazole 1,1-dioxides PMID- 11052132 TI - Endo mode cyclization of 5,6-epoxy-7-octyn-1-ol derivatives PMID- 11052133 TI - Efficient enantioselective synthesis of (R)-(-)-carnitine from glycerol. PMID- 11052134 TI - Asymmetric synthesis of orthogonally protected L-threo-beta-hydroxyasparagine. PMID- 11052135 TI - Oxidative cleavage of indole delta-lactones with m-chloroperbenzoic acid: first synthesis of spiroindolin-2-one gamma-lactones PMID- 11052136 TI - A new substrate for the Biginelli cyclocondensation: direct preparation of 5 unsubstituted 3,4-dihydropyrimidin-2(1H)-ones from a beta-keto carboxylic acid PMID- 11052137 TI - A short, efficient, and stereoselective procedure for the synthesis of cis-3 hydroxymethyl-aziridine-2-carboxylic acid derivatives, important intermediates in the synthesis of mitomycinoids. PMID- 11052138 TI - Facile one-pot preparation of 3-chloro-2-(chloromethyl)propene and an ab initio study of the deamination reaction of nitrosoaziridine PMID- 11052139 TI - Synthesis of cyclic amino acid derivatives via ring closing metathesis on a poly(ethylene glycol) supported substrate PMID- 11052140 TI - The "no reaction" reaction of 1-vinylnortricyclene to tricyclo PMID- 11052141 TI - Molecular structure of a heptadentate cogwheel: C7Me7+ is not planar PMID- 11052142 TI - Recent reviews. 58 PMID- 11052143 TI - Patients and methods. PMID- 11052144 TI - European best practice guidelines 1-4: evaluating anaemia and initiating treatment. PMID- 11052145 TI - European best practice guidelines 5: target haemoglobin. PMID- 11052146 TI - European best practice guidelines 6-8: assessing and optimizing iron stores. PMID- 11052147 TI - European best practice guidelines 9-13: anaemia management. PMID- 11052148 TI - European best practice guidelines 14-16: inadequate response to epoetin. PMID- 11052149 TI - European best practice guidelines 17-18: adverse effects. PMID- 11052150 TI - William Charles Wells (1757-1817) and vestibular research before Purkinje and Flourens. AB - Vestibular research before Flourens typically involved vertigo and eye movements. In 1820 Purkinje integrated these in studies of postrotary vertigo and he is linked with Flourens as a founder of vestibular research. In the late eighteenth century Erasmus Darwin described vertigo in detail, but he did not accept that it involved an oculomotor component. Darwin reached this conclusion despite detailed experiments by William Charles Wells (1757-1817), who described the pattern of postrotary nystagmus and its dependence on head orientation during rotation. Wells generated afterimages prior to rotation and subsequently compared their motions with those of real images. He was able to distinguish between the slow and fast phases of nystagmus, its reducing amplitude following cessation of rotation, its suppression with fixation, and its torsional dimension. In many ways, Wells's experiments were more sophisticated than those of Purkinje, and he should be recognised as a founder of vestibular research. Possible reasons for the neglect of Wells's work are discussed. PMID- 11052151 TI - The effects of repeated optokinetic stimulation on human autonomic function. AB - Numerous animal studies have suggested that the vestibular system modulates respiratory and cardiovascular function. However, relatively few studies have examined vestibular-autonomic interaction in humans. In this study we investigated the effects of repeated horizontal (clockwise or anticlockwise) optokinetic stimulation on systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse and latency to circularvection (CV) in humans, and compared the effects with those of repeated exposure to a stationary visual stimulus. Although all subjects experienced CV, neither mean SBP, DBP or pulse differed significantly between the clockwise/anticlockwise optokinetic stimulation and no optokinetic stimulation conditions. However, SBP and DBP changed significantly over the 20 trials in each test session, even when there was no optokinetic stimulation (P < 0.001 in each case). These results suggest that while horizontal optokinetic stimulation does not significantly affect SBP, DBP or pulse in humans, changes in these variables can be induced by trial repetition itself, even when no optokinetic stimulation occurs. PMID- 11052152 TI - Characteristics of secondary phase post-rotatory nystagmus following off-vertical axis rotation in humans. AB - The nystagmus following yaw earth-vertical axis rotation often reverses direction, a phenomenon known as the "secondary phase". The purpose of this study was to examine the existence and the spatial and temporal properties of the secondary phase of post-rotatory nystagmus following off-vertical axis rotation (OVAR). Eleven normal human subjects were rotated at 120 or 180 degrees/s about an off-vertical axis and stopped in the left ear down or right ear down lateral position. Horizontal and vertical eye positions were recorded with a scleral search coil, and horizontal and vertical slow component eye velocities were computed. Our results indicate that (a) there is a robust secondary phase nystagmus following OVAR, and (b) the direction of the secondary phase nystagmus tends to align with earth-horizontal. These results can be explained by a minor modification of an existing VOR model that has been shown to produce secondary phase responses. PMID- 11052153 TI - Influence of fixation on circular vection. AB - The contribution of fixation to latency of circular vection (CV) was examined in twenty-five normal adults aged 18-30 years. For induction of self-motion a random dot pattern was projected onto a hemispherical dome. The pattern was rotated either about the subject's vertical axis or about their interaural axis at a constant acceleration of 1 deg/s2. For the group tested, the perceived CV latencies were significantly shorter with fixation than without fixation in both horizontal and vertical CV. The effect of fixation was pronounced in subjects with longer latencies. The mean CV latencies for two different fixation points between the subject's eyes and the moving pattern did not differ significantly. Our results suggest that the potential influence of fixation must be carefully controlled in studies of visually induced self-motion. Possible explanations for the effect of fixation on the generation of CV will also be discussed. PMID- 11052154 TI - Difference by instructional set in stabilometry. AB - There is no standard for the awareness of standing posture in stabilometry, yet little research addressing the matter has been carried out. In the present study, we evaluated the influence of different instructional sets during a test on stabilometry. Stabilometry was performed on 349 male subjects. Two different instructions were prepared for the subjects regarding the awareness of their standing posture. These instructions were a) "Please relax when you stand" (R standing), and b) "Please make an effort to minimize your body sway" (E standing). Subjects were classified into four groups according to the combination of these instructions they received. For the five body sway parameters, a comparison between R-standing and E-standing was performed, controlling for possible confounders such as age, height, body weight, educational history, alcohol consumption, and smoking status. The sway length in E-standing was larger than that in R-standing, even after the adjustment for possible confounders. Our results indicate that the difference in the instructional set caused a significant measurement bias. Thorough-going unification of instructions for the stabilometry should be recommended when stabilometry is performed in an epidemiological investigation. PMID- 11052155 TI - Is azithromycin monotherapy as efficacious as cefuroxime plus erythromycin for the treatment of community-acquired pneumonia in hospitalized patients? PMID- 11052156 TI - Which patients with blunt trauma do not require cervical spine x-rays? PMID- 11052157 TI - What is the optimal treatment for lateral ankle ligament ruptures? PMID- 11052158 TI - Which patients with minor head injury do not need computed tomography? PMID- 11052159 TI - Which class of antidepressants is most effective and best tolerated by patients with major depression? PMID- 11052160 TI - Interventions that help victims of domestic violence. A qualitative analysis of physicians' experiences. AB - BACKGROUND: The barriers physicians face when providing care to victims of domestic violence are well detailed in the literature, but few studies provide insight into how physicians overcome these barriers. Our goal was to describe the domestic violence interventions used by physicians who are committed to providing quality health care to battered women. METHODS: We conducted 6 focus groups with 45 San Francisco Bay Area physicians who had intervened with victims of domestic violence. The sessions were audiotaped and transcribed. We constructed, through constant comparison, a template of open codes to identify themes that emerged from the data. RESULTS: Our analysis revealed that physicians viewed validation (ie, providing messages to the patients that they are worth caring about) as the foundation of intervention. Other interventions included labeling the abuse as abuse; listening and being nonjudgmental; documenting, referring, and safety planning; using a team approach; and prioritizing domestic violence in the health care environment. Physicians described a range of rewards for intervening with victims, from seeing a patient change her entire life to subtle shifts in the way a woman thinks of her relationship and herself. CONCLUSIONS: Our study offers insight into how physicians can intervene to help victims of domestic violence. Recent interview and survey studies of battered women support the physician interventions described. PMID- 11052161 TI - Application of the Woman Abuse Screening Tool (WAST) and WAST-short in the family practice setting. AB - BACKGROUND: Our study objectives were to assess the validity and reliability of the Woman Abuse Screening Tool (WAST) in the general population within the family practice setting; to determine the comfort levels of family physicians administering the WAST, their perceptions of its ability to help them identify abused women, and their willingness to continue using it in practice; and to determine the self-reported comfort of patients being asked the WAST questions by their family physicians. METHODS: We included a stratified random sample of 20 physicians practicing in both urban and rural settings drawn from 400 family physicians in London, Ontario, Canada, and the surrounding area. These physicians administered the WAST to 10 to 15 eligible and consenting patients during the course of regular care. Following the physician-patient encounter, patients were asked to complete both a measure about their comfort in being asked each of the WAST questions and the Abuse Risk Inventory (ARI). RESULTS: Scores on the WAST correlated well with those on the ARI. The reliability of the WAST among this sample was demonstrated by a coefficient alpha of 0.75. With the WAST-Short (the first 2 questions of the WAST), 26 of the 307 patients screened (8.5%) were identified as experiencing abuse. The physicians were comfortable administering the WAST to their women patients, and 91% of the patients reported being comfortable or very comfortable when asked the WAST questions by their family physician. CONCLUSIONS: The WAST was found to be a reliable and valid measure of abuse in the family practice setting, with both patients and family physicians reporting comfort with it being part of the clinical encounter. PMID- 11052162 TI - Primary violence prevention. Taking a deeper look. PMID- 11052163 TI - Back-up antibiotic prescriptions for common respiratory symptoms. Patient satisfaction and fill rates. AB - BACKGROUND: In recent years much has been written about the overuse of antibiotics, especially for common respiratory illnesses. One approach to this issue is the use of a back-up prescription, only to be filled if a patient's condition deteriorates or fails to improve. The purpose of our study was to determine patient satisfaction, prescription fill rates, and correlates of these outcomes among patients receiving back-up antibiotic prescriptions. METHODS: In our observational study we obtained survey data from 28 physicians and 2 physician extenders in 3 family practice clinics and their patients presenting with complaints of common respiratory symptoms. We computed patient satisfaction and fill rates of back-up antibiotic prescriptions. Agreement between the perceived need of patients for antibiotics before the office visit and the subjective rating of their physicians of the clinical necessity to prescribe antibiotics for these patients was assessed using the kappa statistic. Finally, we determined correlates of satisfaction and the rate of filling back-up prescriptions. RESULTS: Of the 947 patients enrolled in the study, 46.6% received no antibiotic prescriptions, 30.2% received back-up antibiotic prescriptions, and 23.2% were given immediate-fill prescriptions for an antibiotic. Patients' self reported satisfaction and fill rates for back-up antibiotic prescriptions were 96.1% and 50.2%, respectively. CONCLUSIONS: Our findings indicate that patients were very satisfied with a back-up antibiotic prescription. The fact that half of the patients chose not to fill these prescriptions suggests a potential health care cost savings. PMID- 11052164 TI - Care-seeking behavior for upper respiratory infections. AB - BACKGROUND: Many recent efforts to reduce unnecessary medical services have targeted care of upper respiratory infections (URIs). We tested whether patients who seek care very early in their illness differ from those who seek care later and whether they might require a different approach to care. METHODS: We surveyed by telephone 257 adult patients and 249 parents of child patients who called or visited one of 3 primary care clinics within 10 days (adults) or 14 days (parents) of the onset of uncomplicated URI symptoms. Those who contacted the clinic within the first 2 days of illness were compared with those who made contact later. RESULTS: Although 28% of adults and 41% of parents contacted their clinic within the first 2 days of symptom onset, we found very few differences in the characteristics of the caller or patient between those who called early and later. The illnesses of those who called early were not more severe, and they did not have different beliefs, histories, approaches to medical care, or needs. The only clinician-relevant difference was that adult patients calling in the first 2 days had a greater desire to rule out complications (84.7% vs 64.1% calling in 3 5 days and 70.6% calling after 5 days of illness, P < or = .05). CONCLUSIONS: Those who seek medical care very early for a URI do not appear to be different in clinically important ways. If we are going to reduce overuse of medical care and antibiotics for URIs, clinical trials of more effective and efficient strategies are needed to encourage home care and self-management. PMID- 11052165 TI - Will patients use a computer to give a medical history? AB - BACKGROUND: A patient-entered computerized history, can be used as a means of medical data collection in a large inner city population. We evaluated whether a patient presenting to the Charity Hospital campus of the Medical Center of Louisiana in New Orleans would use a computer to provide medical information. OBJECTIVE: To determine whether patients would self-administer a computerized medical history and find this an acceptable experience DESIGN: A survey questionnaire was given to 100 patients selected from the emergency department walk-in clinic waiting area. SETTING: Charity Hospital emergency room walk-in clinic METHODS: One hundred patients older than 18 years were selected to give a random sample of the population using the Charity Hospital emergency room walk-in clinic for care. The patients received a questionnaire for rating their experience with the computer. Demographics were collected for all patients, including the 13 who declined participation. The main outcome was the patient's perception of the acceptability of using the computerized medical history. A second important outcome measure was patient refusal to participate in the study. RESULTS: Our analysis of the acceptability ratings revealed adequate internal validity (Cronbach alpha=0.75). A single total score was created for these ratings. The participants' scores ranged from 2.0 to 4.0, with a mean of 3.3 (standard error of the mean=0.04). We observed an 83% positive experience in the participating population. CONCLUSIONS: The patients were able to use the computer to enter their medical information. They responded favorably to the experience and appeared to be capable and willing to provide medical information through use of this technology. PMID- 11052166 TI - Alternative pharmacotherapy. Patterns of patient use and family physician practice. AB - BACKGROUND: The use of alternative pharmacotherapies is rapidly increasing. Many persons who use purchased or prepared alternative medications are also cared for by family physicians. We describe patient usage of alternative pharmacotherapies and examine how family physicians handle this in medical practice. METHODS: We recorded data from structured interviews of 178 patients in an academic family medicine practice in a midsized southern city. We then examined the medical records of each participant who reported using some form of alternative pharmacotherapy to determine whether there was discussion of this use with the physician. RESULTS: Approximately one third of the patients reported using some form of alternative pharmacotherapy for 1 year or less, learning about alternative medications mostly from the media, and being generally satisfied with the results. Eighty-four percent of the patients reported not having been asked by their physician about their use of these drugs on the day of their office visit, and more than half reported never having been asked about their use of them. Medical record reviews indicated that for the most part physicians did not document having discussed or making recommendations about the use of alternative pharmacotherapies at any point in their relationship with the patient. CONCLUSIONS: Since many of their patients are using alternative pharmacotherapies, family physicians are encouraged to learn more about what their patients use, to institute easy system-wide changes to facilitate discussion about this use with their patients, to document alternative drugs used, and to give recommendations regarding them. PMID- 11052167 TI - Tympanometry interpretation by primary care physicians. A report from the International Primary Care Network (IPCN) and the Ambulatory Sentinel Practice Network (ASPN). AB - BACKGROUND: The accuracy of data gathered by primary care clinicians in practice based research networks (PBRNs) has been questioned. Tympanometry, recently recommended as a means of improving accuracy of diagnosing acute otitis media, was included as an objective diagnostic measure in an international PBRN study. We report the level of agreement of interpretations of tympanograms between primary care physicians in PBRNs and experts. METHODS: Primary care physicians in PBRNs in the Netherlands, United Kingdom, United States, and Canada enrolled 1773 children aged 6 to 180 months who contributed 6358 tympanograms during 3179 visits. The physicians were trained in the use and interpretation of tympanometry using the Modified Jerger Classification. We determined the level of agreement between physicians and experts for interpretation of tympanograms. One comparison used the 6358 individual ear tracings. A second comparison used the 3179 office visits by children as the unit of analysis. RESULTS: The distribution of expert interpretation of all tympanograms was: 35.8% A, 30% B, 15.5% C1, 12% C2, and 6.8% uninterpretable; for visits, 37.8% were normal (A or C1), 55.6% abnormal (B or C2), and 6.6% could not be classified. There was a high degree of agreement in the interpretation of tympanograms between experts and primary care physicians across networks (kappa=0.70-0.77), age groups of children (kappa=0.69-0.73), and types of visits (kappa=0.66-0.77). This high degree of agreement was also found when children were used as a unit of analysis. CONCLUSIONS: Interpretations of tympanograms by primary care physicians using the Modified Jerger Classification can be used with confidence. These results provide further evidence that practicing primary care physicians can provide high-quality data for research purposes. PMID- 11052168 TI - Family practice research networks. Experiences from 3 countries. AB - Access to data about the clinical problems, patients, and processes that characterise family practice is essential for the development of this specialty. Practice-based research networks (PBRNs) play an increasing role in obtaining these data. We compared 3 PBRNs: one in Wisconsin in the United States, one in Wessex in the United Kingdom, and one in Nijmegen in the Netherlands. We organized our data into 4 key areas for review: the mission of the network, its contribution to the evidence base of family medicine, the management of the network, and the financing of the network infrastructure. Extending the evidence base of family practice is the overriding objective of these networks, and their main focus is on common morbidities. They provide access to unselected patient populations, but there are differences in their size. There are aspects of PBRNs that are common in countries with different health care systems, despite the fact that local circumstances--the research mission or the characteristics of the health care system under which they operate--determine their form and structure. Networks develop over time and their focus and activities may evolve. Financial support for these networks continues to be a problem. PMID- 11052169 TI - How accurate are rapid polymerase chain reaction tests in detecting group B streptococcus colonization in pregnant women? PMID- 11052170 TI - What is the risk of advanced proximal colon neoplasms in patients with and without distal colon disease? PMID- 11052171 TI - Which patients with an ischemic stroke are likely to be helped or harmed by early aspirin treatment? PMID- 11052172 TI - Antibiotics for upper respiratory infections. PMID- 11052173 TI - Antibiotics for upper respiratory infections. PMID- 11052174 TI - A mango for meemo. PMID- 11052175 TI - Enteric bacteria, lipopolysaccharides and related cytokines in inflammatory bowel disease: biological and clinical significance. AB - Ulcerative colitis (UC) and Crohn's disease (CD) [inflammatory bowel disease (IBD)] are both characterized by an exaggerated immune response at the gut associated lymphoreticular tissue level. Such an abnormal and dysregulated immune response may be directed against luminal and/or enteric bacterial antigens, as also supported by murine models of inflammatory bowel disease (IBD) caused by organisms such as Citrobacter rodentium and Helicobacter hepaticus. Bacterial endotoxins or lipopolysaccharides (LPS) have been detected in the plasma of IBD patients and an abnormal microflora and/or an increased permeability of the intestinal mucosa have been invoked as cofactors responsible for endotoxemia. At the same time, the evidence that phagocytosis and killing exerted by polymorphonuclear cells and monocytes and the T-cell dependent antibacterial activity are decreased in IBD patients may also explain the origin of LPS in these diseases. In IBD, pro-inflammatory cytokines and chemokines have been detected in elevated amounts in mucosal tissue and/or in peripheral blood, thus suggesting a monocyte/macrophage stimulation by enteric bacteria and/or their constituents (e.g. LPS). On these grounds, in experimental models and in human IBD, anti-cytokine monoclonal antibodies and interleukin receptor antagonists are under investigation for their capacity to neutralize the noxious effects of immune mediators. Finally, the administration of lactobacilli is beneficial in human IBD and, in murine colitis, this treatment leads to a normalization of intestinal flora, reducing the number of colonic mucosal adherent and translocated bacteria. PMID- 11052176 TI - LPS and LAM activation of the U373 astrocytoma cell line: differential requirement for CD14. AB - CD14 is a membrane protein (mCD14) found on monocytes and neutrophils that is required for the innate immune response to lipopolysaccharide (LPS) and lipoarabinomannan (LAM). CD14 can also be found in serum as soluble CD14 (sCD14) that when bound to bacterial products, enables many non-CD14 bearing cells to be activated. Lipopolysaccharide binding protein (LBP) is a plasma protein that disaggregates and catalytically transfers LPS to CD14. To examine the role of CD14 and LBP in LAM-dependent activation, we used the U373 astrocyte cell line to stably express membrane-bound CD14 (U373-CD14). In serum-free conditions, U373 CD14 cells could respond to LAM stimulation as measured by expression of intracellular adhesion molecule-1 (ICAM-1). Vector control cells (U373-RSV) could not respond to LAM or LPS; but, upon the addition of serum as a source of soluble CD14, control U373-RSV cells could respond to LPS, but not LAM. Therefore, LAM can activate U373 cells only through membrane CD14 and not soluble CD14. We also demonstrate that this membrane CD14-dependent LAM response is greatly facilitated by the addition of LBP. PMID- 11052177 TI - Complement activation by Proteus mirabilis negatively charged lipopolysaccharides. AB - Proteus mirabilis strains are human pathogens responsible for urinary tract infections and bacteremias and may be involved in rheumatoid arthritis. Lipopolysaccharide (LPS, bacterial endotoxin), the major component of the cell wall, is one of the virulence factors of Proteus. In the presented studies, we have investigated complement activation by LPSs isolated from P. mirabilis O10, O23, O30, and O43 strains, which differ in the number of negative COO- groups on their polysaccharide components. Four P. mirabilis strains studied were sensitive to complement-mediated killing, despite complement binding by their LPSs. The optimal complement binding by LPSs was detected in serum with functional assays for both the classical and alternative pathways. Complement activation in 80% serum by the smooth, uronic acid, and hexosamine containing P. mirabilis LPSs was not critically determined by the structure of their O-chain polysaccharides. One of four LPSs used as a model, P. mirabilis O10 LPS, fragmented C3 in an LPS dose- and time-dependent manner. It was detected by crossed-immunoelectrophoresis and capture ELISA with anti-C3c antibodies. The lower complement activation by 023 LPS correlates with its reduced C3 fragmentation, compared with three other Proteus LPSs studied. Rabbit anti-O antibodies enhanced the complement binding and factor C3 fragmentation by O10, O23, O30, and O43 P. mirabilis LPSs. PMID- 11052178 TI - Comparative study of peritoneal macrophage functions in mice receiving lethal and non-lethal doses of LPS. AB - In previous studies, we have observed changes in several functions of peritoneal macrophages from female BALB/c mice with lethal endotoxic shock caused by intraperitoneal injection of Escherichia coli O55:B5 lipopolysaccharide (LPS; 100 mg/kg), which were associated with a high production of superoxide anion and tumor necrosis factor alpha (TNF-alpha). In the present work, both a lethal dose (250 mg/kg) and a non-lethal dose (100 mg/kg) of LPS were used in female Swiss mice. In peritoneal macrophages, the following functions were studied at 2, 4, 12 and 24 h after LPS injection: adherence to substrate, chemotaxis, ingestion of particles, and superoxide anion and TNF-alpha production. In both groups, the results showed a stimulation of adherence, ingestion and superoxide production as well as a decrease of chemotaxis, whereas TNF-alpha could not be detected in either of the two groups. These effects were more evident with the 250 mg/kg dose, especially as regards superoxide anion production, which was higher in the animals treated with a lethal dose of LPS. PMID- 11052179 TI - The inhibitory action of butyrate on lipopolysaccharide-induced nitric oxide production in RAW 264.7 murine macrophage cells. AB - The effect of butyrate, a natural bacterial product of colonic bacterial flora, on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 murine macrophage cells was studied. Butyrate significantly reduced NO production in LPS-stimulated RAW cells. The inhibition was abolished by the removal of butyrate. Butyrate also inhibited the expression of inducible type NO synthase (iNOS) in LPS-stimulated RAW cells. Furthermore, butyrate prevented the activation of nuclear factor (NF)-kappaB through the stabilization of IkappaB alpha and IkappaB-beta. Butyrate did not affect the phosphorylation of mitogen activated protein (MAP) kinases by LPS. It was, therefore, suggested that butyrate down-regulated LPS-induced NO production in RAW cells through preventing the expression of iNOS, and that it was due to the inhibitory action of butyrate on the activation of NF-kappaB. PMID- 11052180 TI - The role of polar and facial amphipathic character in determining lipopolysaccharide-binding properties in synthetic cationic peptides. AB - Two series of peptides, designated K and NK were synthesized and tested for lipid A binding and neutralizing properties. K2, which has an 11-residue amphiphilic core, and a branched N-terminus bearing two branched lysinyl residues does not bind lipid A, while NK2, also with an 11-residue amphiphilic core comprised entirely of non-ionizable residues, and a similarly branched, cationic N terminus, binds lipid A very weakly. Both peptides do not inhibit lipopolysaccharide (LPS) activity in the Limulus assay, nor do they inhibit LPS induced TNF-alpha and NO production in J774 cells. These results are entirely unlike a homologous peptide with an exclusively hydrophobic core whose LPS binding and neutralizing properties are very similar to that of polymyxin B [David SA, Awasthi SK, Wiese A et al. Characterization of the interactions of a polycationic, amphiphilic, terminally branched oligopeptide with lipid A and lipopolysaccharide from the deep rough mutant of Salmonella minnesota. J Endotoxin Res 1996; 3: 369-379]. These data suggest that a clear segregation of charged and apolar domains is crucial in molecules designed for purposes of LPS sequestration and that head-tail (polar) orientation of the cationic/hydrophobic regions is preferable to molecules with mixed or facial cationic/amphipathic character. PMID- 11052181 TI - Endotoxin-mimetic effect of antibodies against Toll-like receptor 4. AB - Monospecific, affinity-purified polyclonal antibodies reacting with the amino terminal half of the mouse Toll-like receptor 4 (Tlr4) ectodomain failed to block LPS effects and, to the contrary, were capable of inducing TNF synthesis when applied to mouse macrophages and cross-linked with a secondary antibody. This effect was observed with macrophages derived from C3H/HeN and C57BL/10ScSn mice, but not with macrophages derived from C3H/HeJ or C57BL/10ScCr mice, indicating a specific, Tlr4-dependent effect. Neither primary nor secondary antibody caused any response if administered in the absence of the other reagent, nor was any response observed in cells from mice lacking Tlr4, or bearing the Lps(d) mutation of Tlr4. These findings support several conclusions. Tlr4, the essential transducer of LPS responses, may act independently of LPS itself. LPS needs not be internalized, nor must it bind to a secondary target within the cell in order to exert its effect; rather, the receptor alone is required for initiation of a signal. The data are consistent with the hypothesis that a conformational change in Tlr4 is required for activation via this receptor, and reveal that the amino terminal half of the Tlr4 ectodomain is a target sufficient for antibody-mediated activation. PMID- 11052182 TI - Minimally invasive approaches to antireflux surgery. AB - Gastroesophageal reflux disease is one of the most common disorders affecting western civilization. Historically, surgical antireflux therapy was reserved for patients who had failed medical therapy, typically in the presence of refractory ulcers or difficult-to-manage strictures. More recently, with improvements in acid control, these acid-pepsin-related complications of reflux have been replaced by the malignant complications of reflux disease, with emphasis now on total control of reflux. Recent developments in surgical technique and the demonstrated effectiveness of a variety of minimally invasive treatment options have changed our approach to these patients. This article summarizes the recommended diagnostic evaluation of patients with reflux symptoms and the current indications for antireflux surgery. The techniques of commonly performed minimally invasive antireflux procedures are described along with a review of the results to be expected. Future prospects for improving the management of reflux are discussed; these include recently described nonsurgical methods for restoring competency to the lower esophageal sphincter. PMID- 11052183 TI - Minimally invasive approaches to acquired shortening of the esophagus: laparoscopic Collis-Nissen gastroplasty. AB - Acquired shortening of the esophagus remains a controversial finding. In some surgical series of patients with gastroesophageal reflux disease, the incidence of clinically significant shortening is low enough that some surgeons have questioned its existence. In the setting of massive hiatial hernia, esophageal shortening has been reported to occur in up to 100% of patients. In association with mild to moderate hiatal hernia, clinically significant esophageal shortening is reported from 2.6% to a much higher percentage of patients, depending on the severity and chronicity of gastroesophageal reflux disease. Failure to recognize this shortening may be responsible for a high failure rate after antireflux surgery. Open Collis gastroplasty is an effective way to manage acquired shortening of the esophagus, and it creates a tension-free intra-abdominal segment of neoesophagus for fundoplication. Minimally invasive approaches to Collis-Nissen procedures have been reported by our group and several others with good short-term results. PMID- 11052184 TI - Laparoscopic repair of giant paraesophageal hernia. AB - Giant paraesophageal hernias (PEHs) account for less than 5% of all hiatal hernias. In contrast to the small type I hiatal hernia, nonsurgical management of giant PEHs may be associated with progression of symptoms and life-threatening complications including hemorrhage, strangulation, and death. Most giant PEHs are associated with a current or previous history of gastroesophageal reflux disease and represent progression of the typical type I hernia to a type III hernia. Conventional open repair is associated with good results and low mortality but also with a significant morbidity and a delay in return to routine activities in this frequently elderly population. Recently, short-term outcome studies have reported that minimally invasive approaches to PEH may be associated with less morbidity, shorter hospital stay, faster recovery, and excellent clinical results. PMID- 11052185 TI - Thoracoscopic and laparoscopic staging for esophageal cancer. AB - Accurate pretreatment staging for patients with esophageal cancer (EC) is becoming increasingly important in the evaluation and comparison of different treatment modalities. Noninvasive staging methods are imperfect in detecting lymph node metastasis in patients with EC. Surgical staging with the thoracoscopic/laparoscopic (Ts/Ls) technique may provide accurate staging information that is useful for evaluating and comparing the results of clinical trials of preoperative chemotherapy and radiotherapy. It can be used to confirm or exclude suspicious distant metastasis found by other staging methods. Pretreatment (lymph node) biopsies obtained by Ts/Ls staging allow further molecular biologic analysis to detect occult lymph node metastasis for more accurate lymph node staging. Since 1992, we have used Ts/Ls staging for EC in 111 patients. We found that Ts/Ls is a promising method for staging lymph nodes in EC patients. A recent study showed that pretreatment surgical lymph node staging can predict response and survival for EC patients receiving trimodality treatment (ie, radiation, chemotherapy, and surgery). The information obtained with surgical staging now offers us the opportunity to optimize therapy to specific patient groups based on the extent of disease at the time of initial presentation. Nevertheless, unlike the practice of mediastinoscopy in lung cancer patients, Ts/Ls staging in EC patients remains an academic interest rather than a clinical practice. The concept of accurate pretreatment staging of EC remains to be realized and accepted in the clinical community. PMID- 11052186 TI - Thoracoscopic and laparoscopic esophagectomy. AB - Esophagectomy is both complex and challenging, and it may be associated with significant morbidity and mortality. With improvements in instrumentation and increasing experience with laparoscopic and thoracoscopic techniques, minimally invasive approaches to esophagectomy are being explored to determine feasibility, results, and potential advantages. Most of this experience has been with case studies or small series, with many surgeons using thoracoscopy in combination with standard laparotomy. Many of the patients have been carefully selected for these procedures because they have small tumors or high-grade dysplasia. Our technique for esophagectomy has evolved from a laparoscopic transhiatal approach to a combined laparoscopic and thoracoscopic approach. Our experience with this procedure has increased, and now we offer this approach to the majority of patients with resectable cancers. We review our operative technique and the results of surgery in our first 50 patients who underwent minimally invasive esophagectomy for cancer or high-grade dysplasia. PMID- 11052187 TI - Laparoscopy or thoracoscopy for achalasia. AB - Achalasia is an esophageal motor disorder of unknown etiology. Typical manometric findings include aperistalsis of the esophageal body coupled with elevated pressure and incomplete relaxation of the lower esophageal sphincter during swallowing. Medical treatments consist of pneumatic dilatation or injections of botulinum toxin. Surgical treatment consists of Heller's myotomy with or without an antireflux procedure. Relief of dysphagia symptoms can be achieved in 85% to 94% of patients undergoing surgical treatment. In the past decade, the minimally invasive approach for the treatment of achalasia has been proven feasible, safe, and effective. We review the role of thoracoscopy and laparoscopy and address controversies in the management of patients with achalasia. PMID- 11052188 TI - Mechanical cardiac assistance: historical perspectives. AB - Cardiac transplantation remains the therapeutic option of choice for treating patients with chronic, progressive, end-stage heart failure. However, over the past 40 years, a number of mechanical assist systems have been developed with the goal of treating and rehabilitating patients with severe circulatory compromise. Today, a wide array of devices is available to provide increasing levels of circulatory support. When used as bridges to heart transplantation, long-term circulatory support systems are a reliable means of keeping heart-failure patients alive as they await suitable donor hearts. Initially, transplant candidates who were receiving this type of support were unable to leave the hospital; today, however, because of advances in mechanical support technology and the portability of the current systems, these patients may return to their homes and even to their jobs while awaiting transplantation. Although heart transplantation may now be considered a routine procedure, the demand for donor hearts will always outweigh the supply, creating a definite need for long-term mechanical circulatory support. Already, clinical trials are underway to test the effectiveness of left ventricular assist devices as long-term support. As smaller, more effective cardiac assist devices become available, they should benefit even more the number of patients who may need permanent circulatory support. The fact that myocardial function can improve enough with chronic ventricular unloading to allow removal of the device may further broaden the use of this technology. PMID- 11052189 TI - Mechanical support for postcardiotomy cardiogenic shock. AB - Postcardiotomy cardiogenic shock (PCCS) results in substantial morbidity and mortality. Despite intraaortic balloon pump and inotropic support, some patients with PCCS continue to have a refractory low cardiac output. For these patients, more effective ventricular assistance is imperative to prevent death. Multiple systems are available for the short-term support of patients with PCCS. Regardless of the device employed, only 25% of these patients survive and are discharged home. Two strategies, however, may improve the outcome of PCCS. One is long-term support by an implantable assist device, which can allow optimal ventricular unloading. Unfortunately, not all cardiac surgery centers offer this type of support. Therefore, the other strategy is the creation of postcardiotomy referral centers that offer long-term support or heart transplantation. Such centers would conserve scarce donor organs, maximize the chance of myocardial recovery, and yield expertise applicable not only to device recipients but also to critically ill heart-failure patients who do not need an implantable pump. PMID- 11052190 TI - Clinical experience with long-term use of implantable left ventricular assist devices: indications, implantation, and outcomes. AB - We describe our clinical experience with 205 implantable left ventricular assist devices at the Cleveland Clinic between December 1991 and January 2000, along with manufacturers' data submitted to the Food and Drug Administration. In patients with end-stage cardiac failure who are suitable candidates for transplantation, these devices serve as excellent bridges to transplantation. Recent modifications have increased pump reliability and reduced thromboembolic rates. The vented electric HeartMate (Thermocardiosystems Inc, Woburn, MA) and the Novacor (Baxter-Novacor, Oakland, CA) left ventricular assist systems allow patients to be discharged from the hospital while awaiting a donor heart. Experience with long-term support is providing insights into permanent implantation of these devices as destination therapy. Although infection remains a major impediment to long-term support, patient-pump interactions leading to changes in the coagulation and immune systems are being recognized, and these interactions may have important implications with respect to thromboembolism, infection, and sensitization to human leukocyte antigens (HLAs). Better understanding of these factors may eventually lead to the development of permanently implantable pumps as an alternative to transplantation. PMID- 11052191 TI - The CardioWest total artificial heart as a bridge to transplantation. AB - The CardioWest total artificial heart (TAH), formerly known as the Jarvik-7 and then the Symbion heart, is the only TAH in current clinical use. A new study, approved by the Food and Drug Administration (FDA), was initiated in 1993 with the goal of approving this pump for commercial release. Since then, 145 CardioWest TAHs have been implanted, including 37 pumps in 36 patients at our center. Our 36 patients were studied prospectively according to the investigational device exemption protocol approved by the FDA. Clinical and hemodynamic data obtained upon patients' entry into the study identified this group as mortally ill. After receiving a CardioWest TAH, 29 of the 36 patients (81%) survived to heart transplantation, and 26 (72% of the total group and 90% of the transplant recipients) have survived for up to 7 years (average, 24 months). Multicomponent anticoagulation, based on readily available tests, and the intrinsic properties of the TAH have resulted in a low linearized stroke rate of 0.48 event per patient-year. There have been no device-related mediastinal infections. In dying patients with nonexistent or severely compromised biventricular function, the CardioWest TAH has proved safe and effective, allowing a 72% survival rate for an average of 24 months. PMID- 11052192 TI - The thoratec ventricular assist device: a paracorporeal pump for treating acute and chronic heart failure. AB - The Thoratec Ventricular Assist Device (VAD) System (Thoratec Laboratories, Pleasanton, CA) is a paracorporeal pump that can provide univentricular or biventricular assistance for patients with heart failure. The system consists of a prosthetic ventricle that has a blood-pumping chamber of Thoralon (Thoratec Laboratories) polyurethane, cannulas for univentricular or biventricular support, and either a hospital-based pneumatic drive console or a portable battery-powered drive unit. For biventricular assistance, 2 pumps are used. The Thoratec voluntary registry indicates that, as of May 2000, this system had been implanted in 1,376 patients, mainly for bridging to transplantation (828 patients) or postcardiotomy support (195 patients); the remaining 353 patients received a hybrid configuration of the device or had incomplete information, so they are not included in this analysis. In the 828 bridge-to-transplant patients, the Thoratec system provided biventricular assistance in 472 cases, left ventricular assistance in 326 cases, and right ventricular assistance in 30 cases for up to 515 days. During the support period, the cardiac index increased significantly from 1.4 +/- 0.8 L/min/m2 to 3.0 +/- 0.5 L/min/m2 (with biventricular assistance and left ventricular cannulation). Sixty percent of the 828 patients underwent transplantation, and the posttransplant survival rate was 86%. In the 195 patients who needed postcardiotomy support, VADs were used for up to 80 days for cardiac recovery. Thirty-eight percent of the patients were weaned from the VAD, and 59% of the weaned group were discharged from the hospital. In addition, 49 postcardiotomy patients were considered for transplantation; of these, 32 received a transplant and 23 were discharged. Patient mobility is being improved by the use of a portable driver. The Thoratec VAD is suitable for a wide range of applications, and efforts are underway to facilitate patient mobility and allow hospital discharge. An intracorporeal version of the VAD, which is currently under development, will help achieve these goals. PMID- 11052193 TI - Future directions of cardiac assistance. AB - During the past decade, mechanical circulatory support has gained increased acceptance as a treatment for patients with severe heart failure who are unresponsive to conventional treatment. Steady progress has been made with respect to technology, patient selection, and postoperative management. Currently, a wide array of circulatory assist pumps offers various levels of assistance and degrees of postoperative mobility. These devices not only save lives (lowering the mortality of heart transplant candidates by 55%) but, in some long-term bridge-to-transplant cases, permit hospital discharge and cardiac rehabilitation. More than 4000 patients have been supported by long-term assist pumps worldwide. In addition to being used for bridging to transplantation or bridging to the use of some other bridging device, long-term circulatory assist devices are being evaluated as bridges to recovery or alternatives to transplantation in selected patients with severe heart failure. Moreover, several total artificial hearts have shown considerable potential in calves and will soon undergo clinical trials aimed at permanent heart replacement. Eventually, as cardiac support or replacement devices become smaller, more durable, and less obtrusive, they may become as conventional and commonplace as pacemakers are today. PMID- 11052194 TI - The plantibody approach: expression of antibody genes in plants to modulate plant metabolism or to obtain pathogen resistance. AB - Immunomodulation is a molecular technique that allows the interference with cellular metabolism or pathogen infectivity by the ectopic expression of genes encoding antibodies or antibody fragments. In recent years, several reports have proven the value of this tool in plant research for modulation of phytohormone activity and for blocking plant-pathogen infection. Efficient application of the plantibody approach requires different levels of investigation. First of all, methods have to be available to clone efficiently the genes coding for antibodies or antibody fragments that bind the target antigen. Secondly, conditions to obtain high accumulation of antigen-binding antibodies and antibody fragments in plants are being investigated and optimized. Thirdly, different strategies are being evaluated to interfere with the function of the target molecule, thus enabling immunomodulation of metabolism or pathogen infectivity. In the near future, optimized antibody gene isolation and expression, especially in reducing subcellular environments, such as the cytosol and nucleus, should turn immunomodulation into a powerful and attractive tool for gene inactivation, complementary to the classical antisense and co-suppression approaches. PMID- 11052195 TI - cDNA cloning and characterization of a plant protein that may be associated with the harpinPSS-mediated hypersensitive response. AB - Hypersensitive response-assisting protein (HRAP) is a novel plant protein that can intensify the harpinPSS-mediated hypersensitive response (HR) in harpinPSS insensitive plants, such as the vegetative stage of sweet pepper. In this report, we identified a HRAP cDNA clone from sweet pepper (Capsicum annuum cv. ECW). The sequence of this cDNA clone showed no appreciable similarity to any other known sequences. However, it contained three positively charged regions, a typical signal peptide and a cAMP-dependent phosphorylation site. The hrap mRNA accumulated preferentially during the incompatible interaction of sweet pepper leaves with a pathogenic bacterium, Pseudomonas syringae pv. syringae. When the hrap gene transcription level was high, the sweet pepper leaves readily expressed the harpinPSS-mediated HR. The hrap gene transcription level in sweet pepper was also higher during the reproductive stage than during the vegetative stage. The HRAP distribution in an individual plant and different plant species was investigated. We found that all the organs of sweet pepper, except fruit, could express two different forms of HRAP. Moreover, the hrap gene was presented in many plant species including tobacco, Arabidopsis, and rice. In conclusion, our results suggest that the hrap gene is widely distributed throughout the plant world and its transcription level correlates with plant sensitivity to harpinPSS. The interaction between HRAP and harpinPSS reveals a novel way to interpret the interaction mechanism between plants and bacterial pathogens. PMID- 11052197 TI - Expression analysis of ESts derived from the inner bark of Cryptomeria japonica. AB - To assist genetic research into Cryptomeria japonica, which is one of the most important forest tree species in Japan, expressed sequence tag (EST) analysis was carried out. The cDNA clones were isolated from a library derived from inner bark tissues. Partial sequences were obtained from 2231 clones, representing 1398 unique transcripts. Putative functions were assigned to 1583 clones, which represented 882 unique transcripts, by a Blast algorithm. Homology analysis suggested that ESTs related to cell wall formation represented about 3% of the clones. Transcripts of plant stress response genes were also abundant in the inner bark library, especially genes involved in wounding and drought responses. This indicates that the stress response systems of this tree species are similar to those of other plants, and that these systems are highly conserved among plant species. The remaining 648 clones, which represented 516 unique transcripts, did not show any significant homology to known sequences in the databases searched: these are expected to represent genes specific to Cryptomeria and, possibly, to related species. PMID- 11052196 TI - Systemic induction of a Phytolacca insularis antiviral protein gene by mechanical wounding, jasmonic acid, and abscisic acid. AB - We have isolated a gene encoding a ribosome-inactivating protein (RIP) from Phytolacca insularis, designated as P. insularis antiviral protein 2 (PIP2). The PIP2 gene contained an open reading frame encoding a polypeptide of 315 amino acids. The deduced amino acid sequence of PIP2 was similar to those of other RIPs from Phytolacca plants. Recombinant PIP2 was expressed in Escherichia coli and was used to investigate its biological activities. Recombinant PIP2 inhibited protein synthesis in rabbit reticulocyte lysate by inactivating ribosomes through N-glycosidase activity. It also exhibited antiviral activity against tobacco mosaic virus (TMV). Expression of the PIP2 gene was developmentally regulated in leaves and roots of P. insularis. Furthermore, expression of the PIP2 gene was induced in leaves by mechanical wounding. The wound induction of the PIP2 gene was systemic. Expression of the PIP2 gene also increased in leaves in a systemic manner after treatment with jasmonic acid (JA) and abscisic acid (ABA), but not with salicylic acid (SA). These results imply that plants have employed the systemic synthesis of the defensive proteins to protect themselves more efficiently from infecting viruses. PMID- 11052198 TI - Plants contain a novel multi-member class of heat shock factors without transcriptional activator potential. AB - Based on phylogeny of DNA-binding domains and the organization of hydrophobic repeats, two families of heat shock transcription factors (HSFs) exist in plants. Class A HSFs are involved in the activation of the heat shock response, but the role of class B HSFs is not clear. When transcriptional activities of full-length HSFs were monitored in tobacco protoplasts, no class B HSFs from soybean or Arabidopsis showed activity under control or heat stress conditions. Additional assays confirmed the finding that the class B HSFs lacked the capacity to activate transcription. Fusion of a heterologous activation domain from human HSF1 (AD2) to the C-terminus of GmHSFB1-34 gave no evidence of synergistic enhancement of AD2 activity, which would be expected if weak activation domains were present. Furthermore, activity of AtHSFB1-4 (class B) was not rescued by coexpression with AtHSFA4-21 (class A) indicating that the class A HSF was not able to provide a missing function required for class B activity. The transcriptional activation potential of Arabidopsis AtHSFA4-21 was mapped primarily to a 39 amino acid fragment in the C-terminus enriched in bulky hydrophobic and acidic residues. Deletion mutagenesis of the C-terminal activator regions of tomato and Arabidopsis HSFs indicated that these plant HSFs lack heat inducible regulatory regions analogous to those of mammalian HSF1. These findings suggest that heat shock regulation in plants may differ from metazoans by partitioning negative and positive functional domains onto separate HSF proteins. Class A HSFs are primarily responsible for stress-inducible activation of heat shock genes whereas some of the inert class B HSFs may be specialized for repression, or down-regulation, of the heat shock response. PMID- 11052199 TI - Antisense suppression of a potato alpha-SNAP homologue leads to alterations in cellular development and assimilate distribution. AB - Using the cDNA-AFLP method, we have isolated a transcript-derived fragment (TDF) which shows a differential expression pattern during tuber organogenesis of Solanum tuberosum L. The TDF was used to isolate a cDNA clone carrying a 1.5 kb insert and potentially coding for a 32.5 kDa peptide which, by homology, represents a potato homologue of an alpha-snap gene and has been designated Stsnap. Northern analysis showed that the Stsnap gene is expressed in actively dividing tissues throughout the potato plant. Analysis of genomic DNA from potato revealed that the Stsnap gene is likely to be a single-copy gene. The expression of antisense Stsnap cDNA under the control of the CaMV 35S promoter results in plants with an altered morphology such as curled leaves. Several of these transgenic lines also display cellular and developmental abnormalities with distinct changes in assimilate transport including accumulation of starch and soluble sugars in source leaves. We argue that these findings are consistent with the hypothetical function of the StSNAP gene product in vesicle targeting and fusion during plant development. PMID- 11052200 TI - The transduction of the signal for grape bud dormancy breaking induced by hydrogen cyanamide may involve the SNF-like protein kinase GDBRPK. AB - Alterations in gene expression during early stages of dormancy release in grapevine buds were analyzed to facilitate the identification of gene products that may mediate the signal transduction of a dormancy-release signal, or derepression of meristematic activity. In the present report we describe the identification of GDBRPK, a transcript for an SNF-like protein kinase that is up regulated upon chemical induction of dormancy release by hydrogen cyanamide (HC). Since SNF and SNF-like protein kinases are known as sensors of stress signals, we hypothesize that GDBRPK may be involved in the perception of a stress signal induced by HC. We also describe a simultaneous and remarkable induction of both PDC and ADH transcripts that was observed shortly after HC application, and was of a transient nature. These data may imply that HC application leads to a transient respiratory stress, which likely results in a temporary increase in the AMP/ATP ratio. Since AMP is known as a stress signal that is sensed by SNF-like kinases, we suggest that the SNF-like GDBRPK could serve as the sensor of this signal. PMID- 11052201 TI - The ternary transformation system: constitutive virG on a compatible plasmid dramatically increases Agrobacterium-mediated plant transformation. AB - This paper describes a so-called ternary transformation system for plant cells. We demonstrate that Agrobacterium tumefaciens strain LBA4404 supplemented with a constitutive virG mutant gene (virGN54D) on a compatible plasmid is capable of very efficient T-DNA transfer to a diverse range of plant species. For the plant species Catharanthus roseus it is shown that increased T-DNA transfer results in increased stable transformation frequencies. Analysis of stably transformed C. roseus cell lines showed that, although the T-DNA transfer frequency is greatly enhanced by addition of virGN54D, only one or a few T-DNA copies are stably integrated into the plant genome. Thus, high transformation frequencies of different plant species can be achieved by introduction of a ternary plasmid carrying a constitutive virG mutant into existing A. tumefaciens strains in combination with standard binary vectors. PMID- 11052202 TI - Characterisation of complementary DNAs from the expressed sequence tag analysis of life cycle stages of Laminaria digitata (Phaeophyceae). AB - Laminariales (Phaeophyceae, Heterokonta) are characterised by a heteromorphic digenetic life cycle with a filamentous, microscopic gametophyte and a highly evolved, macroscopic sporophyte. With the ultimate goal of comparing gene expression in each life cycle stage, complementary DNA libraries were constructed from sporophytes and gametophytes of Laminaria digitata. A set of ca. 500 expressed sequence tags (EST) was generated from each life history phase, by single-run partial sequencing of randomly picked cDNA clones. Comparison of the EST deduced amino acid sequences with database protein sequences assigned a putative identity for 39% of the 412 gametophyte clones and 48% of the 493 sporophyte clones sequenced thus far. These data represent more than 152 different proteins now probably identified in L. digitata. Several of those newly identified proteins are of interest to our understanding of the molecular physiology of kelps, for example their carbon-concentrating mechanisms, cell wall biosynthesis and halogen metabolism. EST analysis also confirmed that genes with long 3'-UTRs are widespread in Laminariales and the study of 5'-UTRs allowed the identification of a Kozak consensus sequence, c(A/C)A(A/C)CAUGGc(G/T). Several potential developmentally regulated differences in gene expression are discussed. PMID- 11052203 TI - Characterisation of two distinct HKT1-like potassium transporters from Eucalyptus camaldulensis. AB - Potassium is an essential macronutrient in higher plants. It plays an important physiological role in stoma movements, osmoregulation, enzyme activation and cell expansion. The demand for potassium can be substantial, especially when the plant concerned is a Eucalyptus tree in excess of 50 m tall. We have isolated two cDNAs, EcHKT1 and EcHKT2, from Eucalyptus camaldulensis (river red gum) which are expressed in leaves, stems and roots. These encode potassium transporter polypeptides with homology to the wheat K+-Na+ symporter, HKT1. EcHKT1 and EcHKT2 both complemented the K+-limited growth of an Escherichia coli K+-uptake deficient triple mutant. EcHKT1 and EcHKT2 also mediated Na+ and K+ uptake when expressed in Xenopus oocytes. A comparison of the EcHKT1 and EcHKT2 sequences and their transport properties indicated that these cDNAs represent two K+ transporters with distinct functional characteristics. The functional and structural conservation between these two E. camaldulensis genes and the wheat HKT1 suggests that they play an important, albeit elusive, physiological role. PMID- 11052204 TI - Protein phosphatase 2A holoenzyme and its subunits from Medicago sativa. AB - We detected an about 200 kDa holoenzyme of protein phosphatase 2A (PP2A) in the crude extract of Medicago sativa microcallus cells by gel permeation chromatography. By polymerase chain reaction (PCR) we isolated two M. sativa cDNA fragments corresponding to the catalytic (C) subunit, and one each coding for the A and the B regulatory subunits of PP2A. The C subunit sequences were different from that published previously, indicating the existence of at least three different isoforms in M. sativa. Using the PCR fragments as probes, we obtained two distinct full-length clones for both the A and B subunits from an alfalfa cDNA library. Our results demonstrate that the components of the PP2A holoenzyme, namely the catalytic and regulatory subunits, are present in alfalfa in several isoforms and that their sequences are highly similar to their plant, yeast and animal counterparts. The distinct regulatory subunit genes are constitutively expressed during the cell cycle. Interestingly, two A-B subunit pairs had parallel mRNA steady-state levels in different plant tissues suggesting that not all of the possible isoform combinations are present in all tissues. The expression of the MsPP2A Bbeta subunit form was induced by abscisic acid indicating a specific function for this protein in the stress response. PMID- 11052205 TI - A transgenic rice cell lineage expressing the oat arginine decarboxylase (adc) cDNA constitutively accumulates putrescine in callus and seeds but not in vegetative tissues. AB - We introduced the oat adc cDNA into rice under the control of the constitutive maize ubiquitin 1 promoter. We studied molecularly and biochemically sixteen independent transgenic plant lines. Significant increases in mRNA levels, ADC enzyme activity and polyamines were measured in transgenic callus. These increases were not maintained in vegetative tissue or seeds in regenerated plants, with the exception of one lineage. This particular lineage showed very significant increases in putrescine preferentially in seeds (up to 10 times compared to wild type and controls transformed with the hpt selectable marker alone). We have demonstrated that in cereals such as rice, over-expression of the oat adc cDNA results in increased accumulation of polyamines at different stages of development. We have also demonstrated that strong constitutive promoters, such as the maize ubiquitin 1 promoter, are sufficient to facilitate heritable high-level polyamine accumulation in seed. Our results demonstrate that by screening adequate numbers of independently derived transgenic plants, it is possible to identify those individuals which express a desired phenotype or genotype. PMID- 11052206 TI - Disability management after stroke: its medical aspects for workplace accommodation. AB - PURPOSE: Return to work (RTW) after stroke is one of the critical issues for both employer and employee. Early RTW is a manifestation of social restoration for the disabled stroke as well as an effective way to reduce social costs related stroke. METHOD: This paper discusses the medical problems referred to RTW after stroke for workplace accommodation. Reviewing the literature, factors influencing RTW after stroke are addressed. RESULTS/CONCLUSION: The process of RTW is extremely individual in each case, and affected by multiple factors. Therefore, it is necessary to individually evaluate precise impact of each factor on RTW. PMID- 11052207 TI - CT/MR imaging: a design tool for custom orthosis. AB - PURPOSE: Orthotics attempts to correct imperfections of the human body. These deficiencies can be either congenital or due to injury. Orthotic devices have continuously evolved over the years and range in sophistication from simple joint braces to spinal body jackets. Generic orthoses, such as knee braces, come in various sizes and can be purchased over-the-counter. Sophisticated orthoses, such as those used to correct scoliosis, have to be custom made. Each of such orthoses has to be fabricated for one particular patient and requires the patient's exact geometry. The conventional technique to acquire patient geometric information has generally been casting. This latter is a time proven method of capturing the geometry of body sections of interest. However, in certain situations, casting might not always be the best alternative to obtain patient geometric data. METHOD/RESULTS: This paper presents a new approach to acquire patient geometric information using CT/MR imaging. The proposed approach can help in cases where casting is not the best option. CONCLUSION: Using CT/MR imaging to acquire patient geometry is shown to be feasible both clinically and economically. PMID- 11052208 TI - Improving functions in leprotic hands. AB - PURPOSE: In the developed countries there is a shift in the rehabilitation strategy from community based services to work site based services for the disabled. Leprosy disabled are more at a disadvantage that many of them have not had the benefit of disability prevention activities and harbour such hand deformities for which the only solution is community based rehabilitation. METHOD/RESULTS: Of the various modalities of rehabilitation the authors consider one for this article suitable for work site based rehabilitation as the Modulan grip-aids. CONCLUSION: Essential components of the material and its uses are described with examples. PMID- 11052209 TI - A synthesis of a vocational assessment system and an information system of technical aids. AB - PURPOSE: In vocational rehabilitation, the selection and tailoring of the work place is tightly bound with the integration or reintegration of people with disabilities into the work environment. Instruments which ease this task are assessment systems as well as information systems of technical aids. To date, these instruments are separated from one another in practice. They also necessitate a broad transfer capacity between the results of an assessment system and the selection of technical aids for a work place design suited for people with disabilities. METHOD/RESULTS: Therefore, when employing assessment system, immediate access to information concerning technical aids would ease the work of experts in the area of vocational rehabilitation. This information should also provide a commentary concerning which aids have been utilized to date in concrete cases of suitably designing work places for people with disabilities. CONCLUSIONS: This article describes the synthesis of a vocational assessment system with an information system of technical aids from the aspect of frequency of use. PMID- 11052210 TI - Accelerating the return to work (RTW) chances of coronary heart disease (CHD) patients: part 1--development and validation of a training programme. AB - PURPOSE: Conventional phase II cardiac rehabilitation (CR) programmes have not resulted in an improvement in returning coronary heart disease (CHD) patients to work in over 35 years. This 4 year field-initiated research, sponsored by the National Institute on Disability and Rehabilitation Research, compares conventional CR programmes with a low-intensity CR programme that simulates elements of work (job-simulated CR programme) in terms of return to work (RTW) and physiological conditioning. The effect of training on physical capabilities of patients participating in the job-simulated CR programme was also of equal interest. METHOD: Thirty patients (15 bypass and 15 angioplasty; 15 males and 15 females) participated in a conventional CR programme (control group). The job simulated CR programme included 15 male and 2 female bypass and angioplasty patients (experimental group). Patients in the control group underwent regular aerobic exercise training (treadmill and bicycle). Experimental group patients participated in a series of low-intensity exercises such as progressive time exercises, flexibility exercises, and dexterity exercises. RESULTS: All patients participating in the low-intensity job-simulated CR programme returned to the same job they held at the onset of myocardial infarction (MI). In contrast, only 60% of the control group patients returned to work; at least one-third of these did not go back to the same job they held at the onset of M1. Patients in both groups achieved the same level of physiological conditioning. The physical functional capabilities of the experimental group patients improved significantly throughout training. CONCLUSION: The results of this field-study lead to the conclusion that a low-intensity phase II cardiac rehabilitation programme that simulates elements of work may be far superior to conventional endurance exercise based cardiac rehabilitation programmes in terms of returning patients to work. Such a programme also strengthens patients, improving their physical capabilities, without compromising their physiological conditioning. PMID- 11052211 TI - Accelerating the return to work (RTW) chances of coronary heart disease (CHD) patients: part 2--development and validation of a vocational rehabilitation programme. AB - PURPOSE: This paper, part 2 of the two-part paper, reviews return-to-work outcomes among individuals with coronary heart disease, who participated in an experimental field study reported in part 1 of the two-part paper. Study results reflected specific job stressors associated with physical and mental demands among various job tasks. Trends suggest that personality characteristics commonly associated with 'type a' personalities and cardiac disease risk factors may also serve as positive forces that influence return-to-work activity. METHODS/RESULTS: Relatively high levels of job satisfaction were reported among most experimental subjects. Despite having high return-to-work expectations, these patients lacked specific strategies and resources to facilitate a concrete return to work action plan. CONCLUSION: The researchers conclude that it is essential for cardiac rehabilitation staff, when creating a return to work transition for their cardiac patients, to explore the physical and psychosocial dimensions of jobs, the receptivity of the employer, and the accommodations needed to promote a safe and timely return to work. PMID- 11052212 TI - Cross-sectional comparison of nerve conduction and vibration threshold testing: do screening tools for occupationally induced cumulative trauma disorders result in differing outcomes? AB - PURPOSE/METHOD: Early diagnosis of Carpal Tunnel Syndrome (CTS) is essential for its effective treatment and subsequent reduction or prevention of the personal and economic losses associated with it. This paper describes a comparison of vibration threshold testing (VTT) and nerve conduction testing (NCT) within the same subjects. RESULTS: Across age and diagnostic category there was little difference between VTT results and the NCT velocity measure in their ability to identify individuals with CTS. CONCLUSION: This is important because VTT is more easily administered and tolerated than NCT and can be used in general screening studies to identify potential CTS cases. PMID- 11052213 TI - Range of motion of the wrist: implications for designing computer input devices for the elderly. AB - PURPOSE: The purpose of this study was to report normative values of the amplitude of joint wrist motions and grip strength for older age groups. METHOD: Volunteers. (N = 147) were divided into four age groups, 60-69, 70-79, 80-89 and 90+ years, with a total of 62 men and 85 women. Maximum range motion values were obtained for wrist flexion, extension and ulnar deviation. In addition, grip strength measures were obtained for each participant. RESULTS: In general, the strength and ROM values for the oldest participants in this study were lower than those of the younger age group (age 60 to 69) and significantly lower than those published for subjects between 25 and 54 years of age. Furthermore, across all age groups males were significantly stronger than females. However, females tended to have greater ROM than males, particularly for wrist extension and ulnar deviations. Joint ROM and grip strength declined significantly with age for both males and females. Comparisons with published data for younger subjects (age 25 35) indicate that a 60-69 year old male, will on average experience a decline in wrist flexion, extension and ulnar deviation of 12%, 41%, and 22% respectively. By age 90, an individual may be expected to have ROM values that are only approximately 60% of an average 30 year old individual. CONCLUSION: The results suggest that the ageing population (particularly men) may face greater difficulty using an input device such as a mouse that relies on motions of the wrist. In addition, the reduced ROM of the wrist may put the elderly at greater risk of developing cumulative trauma disorders. The implications of these findings for the design of input devices are discussed. PMID- 11052214 TI - Levodopa motor complications in Parkinson's disease. AB - Parkinson's disease (PD) is an age-related neurodegenerative disorder with an average onset age of 60 years. In the United States, approximately one million persons suffer from PD, and there are 60,000 newly diagnosed cases every year. The estimated cost of PD to society is $27 billion per year. Based on United States Census Bureau projections, it is estimated that the frequency of PD will increase fourfold by the year 2040, making it an even larger burden on patients, their families and society. PMID- 11052215 TI - Pathophysiology of the basal ganglia in Parkinson's disease. AB - Insight into the organization of the basal ganglia in the normal, parkinsonian and L-dopa-induced dyskinesia states is critical for the development of newer and more effective therapies for Parkinson's disease. We believe that the basal ganglia can no longer be thought of as a unidirectional linear system that transfers information based solely on a firing-rate code. Rather, we propose that the basal ganglia is a highly organized network, with operational characteristics that simulate a non-linear dynamic system. PMID- 11052216 TI - Organization of the basal ganglia: the importance of axonal collateralization. AB - Recent neuroanatomical data obtained with single-axon or single-cell labeling procedures in both rodents and primates have revealed the presence of various types of projection neurons with profusely collateralized axons within each of the major components of the basal ganglia. Such findings call for a reappraisal of current concepts of the anatomical and functional organization of the basal ganglia,which play such a crucial role in the control of motor behavior. The basal ganglia now stand as a widely distributed neuronal network, whose elements are endowed with a highly patterned set of axon collaterals. The elucidation of this finely tuned network is needed to understand the complex spatiotemporal sequence of neural events that ensures the flow of cortical information through the basal ganglia. PMID- 11052217 TI - Anatomy of the dopamine system in the basal ganglia. AB - The dopaminergic nigropallidal and nigrosubthalamic projections control the activity of the globus pallidus and subthalamic nucleus neurons in both normal and pathological conditions. Intrastriatal dopaminergic neurons increase substantially in animal models of Parkinson's disease. They contain GABA, display the ultrastructural features of interneurons and form axo-axonic synapses with putative cortical-like glutamatergic boutons. The local dendritic release of dopamine by neurons in the substantia nigra pars compacta and ventral tegmental also influences basal ganglia functions. Thus, the long-term belief that the effects of dopamine in the basal ganglia were solely mediated through the nigrostriatal system must be changed to take into account extrastriatal dopaminergic projections and intrastriatal dopaminergic neurons. PMID- 11052218 TI - Dopamine receptors: from structure to behavior. AB - The responses obtained with drugs that act at dopamine receptors depend on the spectrum of receptors stimulated, the pattern of stimulation and the neuronal signal-transduction pathways that are activated. In the absence of drugs that reliably discriminate between the various cloned receptors, elucidating the role of these receptors has largely relied on molecular genetic approaches that include expression of genes for receptors in cell lines and manipulation of this expression in animal models. Connecting molecular events that occur consequent to receptor stimulation with the resulting physiological effects entails bridging a complex network of interactions. This article reviews the current understanding of the molecular, cellular and systemic consequences of activation of the different dopamine receptors. PMID- 11052219 TI - Pre- and postsynaptic aspects of dopamine-mediated transmission. AB - Dopamine agonist administration induces changes in firing rate and pattern in basal ganglia nuclei that provide an insight into the role of dopamine in basal ganglia function. These changes support a more complex, integrated basal ganglia network than envisioned in early models. Functionally important effects on basal ganglia output involve alterations in burstiness, synchronization and oscillatory activity,as well as rate. Multisecond oscillations in basal ganglia firing rates are markedly affected by systemic administration of dopamine-receptor agonists. This suggests that coordinated changes in neuronal activity at time scales longer than commonly investigated play a role in the cognitive and motor processes that are modulated by dopamine. PMID- 11052220 TI - Dopamine-mediated regulation of striatal neuronal and network interactions. AB - The dopaminergic system exerts dynamic modulation of glutamatergic afferent drive that is dependent on the temporal pattern of the dopaminergic input and the subtypes of striatal neurons affected. The differences in feedforward inhibition between striatal neurons comprising the direct and indirect output pathway confer distinct response-pattern differences in their respective targets,supporting brief bursts of activity in Type-I neurons but attenuating repetitive activity in Type-II cells. This temporal patterning is further modulated by NO-mediated signaling, and by tonic and phasic dopamine-mediated stimulation, which exerts preferential actions on indirect and direct output neurons, respectively. As a result,the striatal network is forced into state-dependent patterns of activity that differentially regulate muscle tone and voluntary motor activity via distinct output projections from the striatum. PMID- 11052221 TI - Electrophysiology of dopamine in normal and denervated striatal neurons. AB - Parkinson's disease (PD) symptoms originate from the loss of the dopaminergic control of neuronal activity in the striatum. Permanent loss of dopaminergic terminals in the striatum results in an abnormal activity of striatal neurons. The therapeutic treatment with exogenous dopamine, therefore, temporarily restores a balanced synaptic excitation of striatal neurons, counteracting pre- and postsynaptically the excessive glutamate release caused by the degeneration of nigrostriatal dopaminergic fibers. However, chronic treatment is associated with adverse effects that might reflect nonphysiological dopamine replacement. Basic studies on experimental animal models of PD are of crucial importance for the development of therapeutic agents able to provide relief to individuals with PD, without the adverse effects associated with currently available drugs. PMID- 11052222 TI - Molecular effects of dopamine on striatal-projection pathways. AB - Gene regulation studies demonstrate that dopamine differentially regulates the direct and indirect projection neurons of the striatum through their respective expression of the D1 and D2 dopamine receptors. Induction of immediate-early genes (IEGs) in striatal neurons is used to study dopamine-receptor-mediated neuronal plasticity. In the dopamine-depleted striatum there is a switch in receptor-mediated signal transduction mechanisms to produce a supersensitive form of D1- mediated neuronal plasticity. This switch is suggested to underlie dopamine-agonist-induced dyskinetic movements that develop during the treatment of Parkinson's disease. PMID- 11052223 TI - Levodopa-induced dyskinesias and dopamine-dependent stereotypies: a new hypothesis. AB - The basal ganglia are thought to modulate the release or inhibition of movements by way of direct and indirect pathways that act as a push-pull system of cortico basal ganglia circuits. Here we suggest a three-pathway model of the basal ganglia that takes into consideration the fundamental division of the striatum into striosomes and extrastriosomal matrix. We suggest that, in addition to the balance between direct and indirect pathways on which normal release of individual movements depends, the balance of activity between these matrix-based pathways and the striosomal pathway regulates the frequency of release of given behavioral sequences and, thus, modulates behavioral focus. Differential plasticity in these compartmentally organized circuits might contribute to the development of L-dopa-induced dyskinesias under parkinsonian conditions and dopamine-receptor-agonist induced stereotypies under normal conditions. PMID- 11052224 TI - Metabolic effects of nigrostriatal denervation in basal ganglia. AB - In the past, functional changes in the circuitry of the basal ganglia that occur in Parkinson's disease were primarily analyzed with electrophysiological and 2 deoxyglucose measurements. The increased activity of the subthalamic nucleus (STN) observed has been attributed to a reduction in inhibition mediated by the external segment of the globus pallidus (GPe), secondary to the loss of dopaminergic-neuron influence on D2-receptor-bearing striato-pallidal neurons. More recently, in situ hybridization studies of cytochrome oxidase subunit I have confirmed the overactivity of the STN in the parkinsonian state. In addition, this technique has provided evidence that the change in STN activity is owing not only to decreased inhibition from the GPe but to hyperactivity of excitatory inputs from the parafascicular nucleus of the thalamus and the pedunculopontine nucleus in the brainstem. PMID- 11052225 TI - Striatal dopamine- and glutamate-mediated dysregulation in experimental parkinsonism. AB - Characteristic changes involving interactions between dopamine and glutamate in striatal medium spiny neurons now appear to contribute to symptom production in Parkinson's disease (PD). The balance between kinase and phosphatase signaling modifies the phosphorylation state of glutamate receptors and thus their synaptic strength. Sensitization of spiny-neuron NMDA and AMPA receptors alters cortical glutamatergic input to the striatum and modifies striatal GABAergic output, and thus motor function. Conceivably, the pharmacological targeting of spiny-neuron mechanisms modified in PD will provide a safer and more effective therapy. PMID- 11052226 TI - Dopamine-receptor stimulation: biobehavioral and biochemical consequences. AB - The MPTP monkey is a well-characterized animal model of parkinsonism and provides an exceptional tool for the study of dyskinesias induced by dopamine-like agents. Several such agents have been tested during the past 15 years, and it has been found that the duration of action of these compounds is the most reliable variable with which to predict their dyskinesiogenic profile. It is proposed that L-dopa-induced dyskinesias represent a form of pathological learning caused by chronic pulsatile (nonphysiological) stimulation of dopamine receptors, which activates a cascade of molecular and biochemical events. These events include defective regulation of Fos proteins that belong to the deltaFosB family, increased expression of neuropeptides, and defective GABA- and glutamate-mediated neurotransmission in the output structures of the basal ganglia. PMID- 11052227 TI - PET studies and motor complications in Parkinson's disease. AB - Parkinson's disease (PD) patients with motor complications show a greater reduction in putamen [18F]dopa uptake on positron emission tomography (PET) compared with sustained responders to L-dopa, although individual ranges overlap considerably. This implies that, although loss of putamen dopamine storage predisposes motor complications in PD, it cannot be the only factor determining timing of onset. Additional PET studies suggest that loss of striatal dopamine storage capacity along with pulsatile exposure to exogenous L-dopa results in pathologically raised synaptic dopamine levels and deranged basal ganglia opioid transmission.This, rather than altered dopamine receptor binding, then causes inappropriate overactivity of basal ganglia-frontal projections, resulting in breakthrough involuntary movements. PMID- 11052228 TI - Continuous dopamine-receptor stimulation in advanced Parkinson's disease. AB - Intermittent or pulsatile dopamine-receptor stimulation is postulated to induce plastic changes in motor systems that are responsible for the development of the motor fluctuations and dyskinesia that complicate long-term L-dopa therapy of Parkinson's disease. As a corollary to this hypothesis, continuous dopamine receptor stimulation can avoid or reverse these complications. Such continuous stimulation is unlikely to mimic completely the normal function of the dopaminergic system, but should avoid the supra-physiological swings in extracellular dopamine that accompany intermittent L-dopa dosing. The concern is that this continuous stimulation might induce tolerance rather than sensitization to some effects of L-dopa. Open clinical trials support the value of continuous dopaminergic stimulation in Parkinson's disease with established motor complications, but rigorous studies, although experimentally difficult, are needed. PMID- 11052229 TI - Continuous dopamine-receptor stimulation in early Parkinson's disease. AB - Chronic L-dopa therapy is associated with the development of motor complications in the majority of Parkinson's disease (PD) patients. Although the precise mechanism responsible for these events is not known, increasing laboratory and clinical evidence points to a sequence of events that is initiated by abnormal pulsatile stimulation of dopamine receptors by the intermittent administration of agents with short half-lives such as L-dopa. Initiating therapy with a long acting dopamine agonist has been shown to delay the onset and reduce the severity of motor complications in MPTP monkeys and PD patients. Administering L-dopa with a catechol-O-methyltransferase (COMT) inhibitor to block its peripheral metabolism increases its plasma half-life and might have a similar effect. Thus, a rational strategy for treating PD would be to initiate therapy with a long acting dopamine-receptor agonist and supplement at the appropriate time with L dopa combined with a COMT inhibitor. PMID- 11052230 TI - Olmsted syndrome: report of two new cases and literature review. AB - Olmsted syndrome is a rare keratinization disorder; 18 cases have been published so far. It associates a mutilating cogenital palmoplantar keratoderma with periorificial erythematokeratotic lesions. We report herein two new unrelated male children with Olmsted syndrome (OS), one of whom was studied by light and electron microscopy. Our histological, immunohistochemical, and ultrastructural findings suggest that this disease is related to epidermal hyperproliferation. We present herein a review of the twenty cases published so far and discuss the major clinicopathological and genetic features of this disease. PMID- 11052231 TI - Is renal function altered in patients with psoriasis vulgaris?--A short review. AB - Psoriasis is a chronic inflammatory dermatosis with distinct microvascular changes. Although it is generally accepted that the psoriatic process is limited to the skin, it is not excluded that similar vascular lesions might be present in internal organs, such as the kidneys. This review summarizes data on renal function in psoriatic patients who were never treated with the potentially nephrotoxic drugs used for treatment of psoriasis. The limited number of such studies is mainly concentrated on microalbuminuria. Enhanced urinary albumin excretion at the level of microalbuminuria has been found in some psoriatic individuals. All other routine laboratory renal tests were within their normal ranges. As microalbuminuria is regarded as a subclinical marker of glomerular dysfunction, the authors hypothesize that some psoriatic patients may present subclinical glomerular changes. However, kidney histopathology is necessary to confirm this hypothesis. PMID- 11052232 TI - Soluble thrombomodulin, a possible predictor of thrombotic crisis in thrombotic disease. AB - Serum soluble thrombomodulin is known to be a factor in visceral vascular disorders such as organic vasculitis in collagen diseases, but recently it has also been reported as a predictor of thrombotic crisis in thrombotic diseases. In this report, serum soluble thrombomodulin levels and events of thrombotic crises in clinical patients were studied retrospectively in our dermatology department over the past five years. I found an increase in soluble thrombomodulin one to two months before the crisis in eight of ten patients including three with anti phospholipid syndrome, two with lupus anticoagulant-positive systemic lupus erythematosus, four patients with chronic disseminated intravascular coagulation syndrome and one patient with thrombotic thrombocytopenic purpura. A decrease was found after treatment. Other tested parameters did not respond as soluble thrombomodulin, and they were not useful for predicting the crisis one to two months before the crisis. These results suggest the possibility that soluble thrombomodulin might be an important factor in predicting thrombotic crisis. PMID- 11052233 TI - Lichen planus: a clinical and epidemiological study. AB - Clinical and epidemiological data from 232 patients with lichen planus is presented. Lichen planus constituted 0.38% of the total dermatology, outpatients diagnosed. The patient ages ranged from 8 to 76 years, most being in the age range from 20 to 49 years. Duration of disease varied from 1 month to 7 years. Both sexes were equally affected. The majority of the patients (47.4%) had classical lesions followed by hypertrophic and actinic lichen planus next in frequency. Itching was the predominant symptom in 79.3%. Limbs were the most frequent initial site of onset (55.6%). Mucosal involvement along with cutaneous lesions were observed in 16.8% and genital involvement in only 5.2%. Nail changes were observed in 15.1% of patients. A history of recurrence of the disease was obtained from 10.3% of patients. Liver disease was found to be associated in 2.2% of patients. No malignant changes were observed in any of the lesions of lichen planus. PMID- 11052234 TI - Small congenital nevi associated with melanoma: case reports and considerations. AB - Melanocytic nevi, both congenital and acquired, are considered to be precursors of melanomas. Data about the malignant potential of these nevi are conflicting, particularly with reference to the nevus of the smallest size. Patients with preexisting melanocytic nevi (both congenital and acquired) have risks of developing melanoma that differ from those of subjects without them. The purpose of this study was to verify the presence of melanoma in preexisting nevi both congenital (congenital nevus associated melanoma) (CNAM) and acquired (ANAM). In particular, we investigated melanomas associated with small congenital nevi (SCN). A cohort of 190 patients with primary melanomas was studied. Congenital nevi were called "small" (SCN) when their diameters were less than 1.5 cm. Epiluminescence microscopy (ELM) was performed to further improve the clinical diagnosis and to observe the more subtle changes in the preexisting nevi. Forty of the 190 cases of melanoma were associated with preexisting nevi; of these, 15 had congenital features with a CNAM largest diameter of 1.5 cm. These 15 cases were melanomas of the superficial type with a mean tumor thickness lower than that of ANAM (0.33 vs 1.50). There were no differences between the locations of CNAM and other melanomas. Male patients were significantly more affected. ELM microscopy permitted us to detect the early malignant changes in nevi and thus to improve our diagnosis. A high percentage of small congenital nevi were found to be associated with melanomas. They may be considered as melanomas precursors. Because of their large number and frequency, prophylactic removal of all SCN is not feasible. However, they should be removed as soon as possible when clinical or ELM changes are observed. PMID- 11052235 TI - A case of Dowling-Degos disease suggesting an evolutional sequence. AB - We report a 47-year-old woman who presented with asymptomatic reticulate hyperpigmentations on the neck, lateral face, axillae, trunk, inguinal areas, and dorsa of both hands and feet. We thought it was an unusual case in the spectrum between the pole of Dowling-Degos disease (DDD) and that of reticulate acropigmentation of Kitamura (RAK). Another interesting point was that the biopsied specimens from the abdomen, neck, and axillary lesions showed somewhat different histopathologic features from typical DDD, suggesting an evolutional sequence. From these findings we suggest that a lichenoid inflammation may be responsible for the typical maculo-papular lesions of DDD. PMID- 11052236 TI - Two cases of confluent and reticulate papillomatosis: successful treatments of one case with cefdinir and another with minocycline. AB - The present report presents two cases of confluent and reticulate papillomatosis. Case 1 was a 24-year-old man who had suffered from skin eruptions for six months, and Case 2 was a 19-year-old woman who had had this disease for three days. In both patients, reticular dark brown papules, accompanied by mild keratosis and infiltration, spread from the trunk to the neck and upper arm. Direct light microscopy did not detect the presence of any fungi, and histopathological examinations confirmed hyperkeratosis, acanthosis, papillomatosis, and mild small round-cell infiltration. Thus, these patients were diagnosed as confluent and reticulate papillomatosis. Neither one had diabetes or thyroid dysfunction. In Case 1, cefdinir was effective, and in Case 2, minocycline hydrochloride and ketoconazole were effective. To the best of our knowledge, this was the first documented case of confluent and reticulate papillomatosis responding to cefdinir. PMID- 11052237 TI - Acrodermatitis enteropathica-like eruption in an infant with nonketotic hyperglycinemia. AB - Acrodermatitis enteropathica is a rare inherited disorder characterized by zinc deficiency and a triad of dermatitis, diarrhea, and alopecia. It is an autosomal recessive condition thought to be due to the inability to absorb zinc from the gastrointestinal tract. Acquired zinc deficiency due to a variety of etiologies may produce a similar clinical picture. These causes include inadequate supply, malabsorption, and low zinc stores. In addition to zinc, deficiencies of other nutrients such as branched chain amino acids have induced an acrodermatitis enteropathica-like eruption. We describe a case of a 26-month-old boy with a rare inborn error of metabolism known as nonketotic hyperglycinemia who developed an acrodermatitis enteropathica-like eruption. In addition to zinc deficiency, the patient was deficient in branched chain amino acids due to a low protein diet instituted to reduce his elevated glycine levels. The rash did not respond to zinc replacement alone, and therefore is most likely a combination of amino acid and zinc deficiency. Acrodermatitis enteropathica-like eruptions have been described in other conditions that cause decreased serum amino acids, such as maple syrup urine disease and organic acidurias. This is the first case describing an association between acrodermatitis enteropathica and nonketotic hyperglycinemia. PMID- 11052238 TI - Three unusual siblings with Harlequin icthyosis in an Indian family. AB - Harlequin fetuses occurring as three siblings in an Indian family are described here. All three were preterm, low birth weight, and did not survive. There was no history of consanguinous marriage in the parents or in the family. Thus autosomal recessive inheritance appears to be a remote possibility, although not impossible or, as recently described, these recurrent harlequin fetuses could be the result of new dominant mutations with parental mosaicism. PMID- 11052239 TI - A case of intramuscular hemangioma of the back. AB - Intramuscular hemangioma is an uncommon tumor that usually involves an extremity. They rarely occur after the age of 30. We report a case of an intramuscular hemangioma occurring on the back of a 39-year-old man. Histopathologically, the tumor consisted mainly of large vessels between the muscle fibers. Phleboliths were observed in some of the vessel walls. PMID- 11052240 TI - A case of lichen planus-like keratosis: deposition of IgM in the basement membrane zone. AB - An 81-year-old woman presented with a round erythematous macule with keratotic scales on her left hand. The skin specimen histologically showed hypergranulosis and apoptotic keratinocytes in the epidermis with lichenoid infiltration of lymphocytes. Parakeratosis seen in the hyperkeratotic cornified layer indicated lichen planus-like keratosis, as distinguished from lichen planus. Direct immunofluorescence study revealed the linear deposition of IgM in the basement membrane zone; IgG, IgA and C3 were not detected. PMID- 11052241 TI - Lichen planus after HBV vaccination in a child: a case report from Nepal. AB - Vaccination against hepatitis B virus has rarely been associated with lichen planus. We report a case of this kind in a child from Nepal. A 12-year-old boy had developed generalized itchy violaceous papules and plaques six weeks after the second dose of hepatitis B virus vaccine. Serum HBsAg and HBeAb were negative, but HBsAb was positive. New crops of generalized, similar eruptions developed after the booster dose of vaccine. All the lesions resolved within three months of systemic steroid therapy. There was no recurrence after one year of follow up. Awareness of such an association is necessary, especially in children, because vaccination campaigns are increasing. PMID- 11052242 TI - Lichen simplex chronicus after herpes zoster. PMID- 11052243 TI - Influence of volatile and intravenous anesthetics on the threshold of the acoustically evoked stapedius reflex. AB - The influence of volatile and intravenous anesthetics on the threshold of the acoustically evoked stapedius reflex (SR) was studied prospectively in 45 patients undergoing elective ENT surgical procedures. After premedication with flunitrazepam the patients were randomly assigned to one of nine groups. Group I: 70% nitrous oxide (N2O) in oxygen (O2); Groups II-VII: induction of anesthesia with intravenous thiopental, followed by mask inhalation with 100% O2 and 1.13% halothane (Group II), 2.52% enflurane (Group III) or 1.73% isoflurane (Group IV); or 70% N2O in oxygen, and 0.44% halothane (Group V), 0.86% enflurane (Group VI) or 0.75% isoflurane (Group VII): Group VIII: intravenous midazolam and ketamine; and Group IX: intravenous midazolam and alfentanil. Tympanometry and ipsilateral and contralateral SR measurements were performed when the effects of the anesthetics had achieved a steady state. Flunitrazepam raised the SR threshold only slightly. Substances applied during inhalation anesthesia either markedly increased the threshold contralaterally more than ipsilaterally (thiopental, N2O), or suppressed the reflex completely (thiopental, all volatile anesthetics with or without N2O). Under intravenous anesthesia the reflex was always present. The midazolam-ketamine combination influenced the threshold bilaterally only slightly, while the midazolam-alfentanil combination led to a pronounced, contralaterally significant elevation of the threshold. Based on its minimal influence on the SR threshold, flunitrazepam is especially suitable for sedation and the midazolam-ketamine combination for anesthesia in audiological diagnostic procedures. PMID- 11052244 TI - Preventive magnesium supplement reduces ischemia-induced hearing loss and blood viscosity in the guinea pig. AB - The effect of magnesium (Mg) on ischemia-induced hearing loss was investigated in two groups of adult pigmented guinea pigs of either an optimal or suboptimal (physiologically high or low) Mg status maintained by different diets. Total Mg concentrations of the perilymph, cerebrospinal fluid, blood plasma and red blood cells were found to differ significantly between the two groups, as tested in a previous study. Local vascular impairment was produced by unilateral ferromagnetic thrombosis of cochlear blood vessels. Cochlear blood flow (CBF) and hearing function were measured using laser Doppler flowmetry and auditory brain stem response audiometry, respectively. Ferromagnetic thrombosis resulted in significant reductions of the mean apical CBF in both experimental groups and of the mean basal CBF in the low Mg group compared to the contralateral ears. In the high Mg group, the basal CBF was not decreased. However, the laser Doppler signals revealed considerable interindividual variations and the differences found between the two experimental groups were not significant. In contrast, the hearing loss in the low Mg group was significantly higher than that in the high Mg group. A correlation was found to exist between the vascular impairment and the hearing threshold shift. In a separate series, we also tested the effect of Mg on hemorheology and found both the blood viscosity and blood viscoelasticity to be significantly lower in the high Mg group than in the low Mg group, depending on the shear rates tested. The present findings show that a preventive oral Mg supplement can significantly reduce the rate of ischemia-induced hearing loss and improve blood viscosity in the guinea pig. PMID- 11052245 TI - Preliminary results of a new delivery system for gentamicin to the inner ear in patients with Meniere's disease. AB - Intratympanic gentamicin therapy has gained some clinical popularity in the treatment of vertigo associated with Meniere's disease. This therapeutic modality offers some advantages over traditional surgical treatment. The vestibulotoxic effect of gentamicin is well documented, but there is no general agreement about the dose needed to control vertigo attacks without affecting hearing. In the current preliminary study 27 patients with Meniere's disease refractory to medical management were treated by small doses of gentamicin delivered via microcatheter in the round window niche and administered by an electronic micropump. The patients received a total dose of 0.24-90 mg. The effect on vestibular symptoms resulted in the cessation of vertigo in the 22 patients, control of drop attacks in 4 of 6, and release of aural pressure and fullness in 2 of 4. Significant hearing loss (anacusis) occurred in six patients, slightly related to the flow rate in the pump setting. Different explanations for the loss of hearing are presented. The new delivery system for gentamicin appears to be effective in controlling vertigo, but with an unacceptable negative effect on hearing. The effectiveness and the safety of this new delivery system must be investigated further in controlled studies. However, it opens up the possibility of future novel ways of treating inner ear diseases, such as sudden deafness and tinnitus, as well as for the protection, repair, and regeneration of inner ear sensory cell function in damage due to aging, noise, toxic substances, immune reactions, etc. PMID- 11052246 TI - Otoscopic findings in relation to tympanometry during infancy. AB - To determine the value of different otoscopic findings in diagnosing otitis media with effusion, 3,780 otological examinations of 250 infants at ages 0-2 years were compared with tympanometric findings. Otoscopy was performed by an experienced otologist and research-audiologist prior to tympanometry. The following success rates for these procedures were found: otoscopy, 85-91%, tympanometry, 74-94%, either otoscopy or tympanometry, 89-99%. "Glue" was noted as the most frequent abnormal otoscopic feature, i.e., depending on age, 6-32%. Fluid line/ bubbles, bulging/hyperaemia or otorrhoea were rarely (0-5%) observed and contributed little to the diagnosis of otitis media with effusion. No structural changes of the tympanic membrane were observed. Analysis of tympanometric data revealed that the prevalence of tympanogram types B (21-44%), C1 (0.5-16%) and C2 (0-8%) was age-dependent. Comparison of otoscopic and tympanometric findings indicated sufficient diagnostic agreement (Cohen's kappa between 0.41 and 0.74) at age 6-24 months. In conclusion, otoscopy appears to be a valid instrument for diagnosing otitis media with effusion, at least from 6 to 24 months of age. In addition, the results of our study suggest that tuba dysfunction and otitis media with effusion are age-dependent disorders during infancy, without structural effects on the tympanic membrane. PMID- 11052247 TI - Postural control in horizontal benign paroxysmal positional vertigo. AB - Sixteen patients affected by benign paroxysmal positional vertigo of the horizontal semicircular canal (BPPV-HSC) were investigated by means of dynamic posturography (DP) and during bithermal caloric stimulation. Data were compared to data from 40 patients with benign paroxysmal positional vertigo of the posterior semicircular canal (BPPV-PSC) and 20 healthy controls. No postural deficit was observed before or after a liberative Lempert's manoeuvre when patients were compared to control subjects. BPPV-PSC postural scores were significantly impaired compared to scores from the BPPV-HSC group. A residual significant postural impairment was also observed after a successful liberative manoeuvre in the BPPV-PSC group. Electronystagmographic recordings before recovery revealed significant hypoexcitability of the affected ear in 8/16 patients of the BPPV-HSC group. After the liberative manoeuvre, a symmetric bilateral response to caloric stimulation was recorded in all patients. Three main conclusions can be drawn from the present data. First, disorders of the horizontal semicircular canal do not change postural control. Second, dynamic posturography can detect the postural imbalance due to posterior semicircular canal dysfunction even after resolution of paroxysmal vertigo attacks. Third, utricular dysfunction can be ruled out as a cause of the residual postural deficit observed in BPPV-PSC patients. Therefore the recovery delay observed even 1 month after the liberative manoeuvre in the BPPV-PSC-group might be due to the persistence of small amounts of residual debris in the canal, to paralysis of ampullar receptors, or to the time needed for central vestibular re-adaptation. PMID- 11052248 TI - Epithelial-myoepithelial carcinoma of the nasal cavity. AB - A 22-year-old male presented with a 1-year history of nasal obstruction due to a polypoid mass in the right nasal cavity. Histopathologic examination revealed the tumor to consist of a mixture of a trabecular structure with a double-layered arrangement of inner dark cells and outer clear cells. Immunohistochemical examination showed the clear cells to be positive for alpha-smooth muscle actin and S-100 protein. Ultrastructural examination confirmed the myoepithelial cell origin. The tumor was excised and no recurrence or metastasis was found 40 months after surgery. We describe here a rare case of epithelial-myoepithelial carcinoma arising from the nasal cavity, one of the most unusual locations. PMID- 11052249 TI - Choanal polyp of sphenoidal origin. AB - Sphenochoanal polyp is a rare entity which originates in the sphenoid sinus cavity and extends into the choana via the ostium. It presents in a similar manner to the more common antrochoanal polyp. Radiological examination is necessary to differentiate between these two types. We present a case of sphenochoanal polyp and review the clinical, radiological and pathological features. The role of endoscopic sinus surgery is emphasised. PMID- 11052250 TI - Management of subglottic stenosis in infancy and childhood. AB - During the 12-year period between June 1987 and June 1999, 141 children underwent curative treatment for subglottic laryngeal stenosis at La Timone Children's Hospital in Marseille, France. Ninety-six children (68%) were under the age of 5 years; 106 (75%) presented with acquired stenosis and 93 had narrowing involving over 70% of the subglottic lumen. Endoscopic laser surgery was performed in 25 cases and open surgery in 116. Open surgical techniques included laryngotracheoplasty with autologous cartilage interposition in 83 cases, laryngotracheal split in 22, and cricotracheal resection in 11. After decannulation, 132 children (94%) were able to breathe normally through the upper airway. Perspectives for development of new techniques and improvement of conventional methods are discussed. PMID- 11052251 TI - Partial epiglottectomy as a possible treatment for globus pharyngeus? AB - Globus pharyngeus is a relatively common complaint in an ear, nose, and throat consulting room and may account for 3-4% of outpatient referrals. The cause is still unknown, although a number of hypotheses have been suggested. Between 40% and 75% of the patients remain symptomatic despite any treatment regimen. Thirteen patients from a group of 124 with the diagnosis of globus pharyngeus and no response to medical treatment were treated with partial epiglottectomies. One year after the surgery all but one patient were free of symptoms. Our experience indicates that partial epiglottectomy can be a good treatment for those patients with globus pharyngeus in whom no cause is found after all studies are performed or when medical treatment fails. PMID- 11052252 TI - Histopathological predictors of occult lymph node metastases in supraglottic squamous cell carcinomas. AB - Lymph node metastasis appears to be the most important factor determining survival in patients with squamous cell carcinoma of the larynx. Supraglottic laryngeal carcinomas have a known tendency to metastasize to cervical lymph nodes because of the extensive lymphatic network present. This retrospective cohort study was conducted to define possible histopathological parameters affecting cervical lymph node metastasis and then using these parameters to create a scale to predict occult lymph node metastasis in supraglottic squamous cell carcinoma. The pathological slides of 61 operated patients were reevaluated for tumor grade, lymphatic-vascular invasion, invasion pattern of tumor margins, perineural invasion and lymphocytic infiltration. Grade (P < 0.001), lymphatic-vascular invasion (P < 0.001) and tumor margins (P = 0.007) were found to be closely associated with neck metastasis. To define the risk factors for occult metastasis, a grading scale was created by using grade (G), lymphatic-vascular invasion (L) and tumor margin (M) findings of patients. None of the patients with a GLM value of zero developed occult metastasis. On the other hand occult metastasis was found in 58.8% of N0 patients with a GLM value that was more than zero. These findings indicate that patients with high-grade tumors having infiltrating borders and lymphatic-vascular invasion have a high risk for occult metastasis so that elective treatment of the neck either by neck dissection or radiotherapy should be added to therapy. Serial sections of specimens are needed to avoid missing metastatic loci of disease. PMID- 11052253 TI - Fish bone inhalation in infancy. AB - Inhalation of sharp bones by infants is an uncommon but hazardous occurrence. management relies not only on a high index of clinical suspicion and timely treatment but also demands expertise in therapeutic bronchoscopy. The potential complications and the management of this condition are reviewed. PMID- 11052254 TI - Neurofibroma of the auriculotemporal nerve. AB - Despite the extensive branching of the trigeminal nerve, solitary neurofibromas along its branches are a rare finding. We report our management of a neurofibroma of the right auriculotemporal nerve in a 46-year-old women. A chain of small nodules palpable in the right postauricular region was associated with increasing pain radiating into the postauricular and temporoparietal regions of her head. Magnetic resonance imaging and computed tomography showed several small ovoid lesions extending from the postauricular region to the infratemporal fossa. The lesions were removed surgically. The facial nerve adhered to the dorsal side of the largest nodule, but this could be removed without sequelae. The auriculotemporal nerve was identified as the nerve of origin and was removed together with the lesions. Histopathological examination was consistent with a neurofibroma with early plexiform cell formations. Clinical findings are discussed. PMID- 11052255 TI - Lymphepithelioma-like carcinoma of the lacrimal gland. AB - In this report a patient with a lymphoepithelioma (LE)-like carcinoma of the lacrimal gland is described for the first time in the literature. LE-like carcinomas outside the nasopharynx rarely occur in the major and minor lacrimal glands of natives of Greenland, Inuit or natives of southern China. The patient's tumor was extirpated using a Kronlein approach followed by total parotidectomy and modified radical neck dissection on the ipsilateral side after the detection of suspicious lymph nodes by ultrasound transmission. Adjuvant radiochemotherapy with cisplatin and 5-fluorouracil was then carried out. Three years later there is no sign of recurrence. As a result of this case we recommend careful examination of the orbit and lacrimal gland in cases of LE-like cancer with an unidentified primary tumor. PMID- 11052256 TI - Schwannoma of the nasal septum: a case report with review of the literature. AB - Schwannomas are neurogenic neoplasms rarely found in the sinonasal tract, where localization to the nasal septum is exceedingly rare (only 11 cases have been described in the western literature). We report the case of a 29-year-old white male with a schwannoma completely filling the left nasal fossa and arising from the bony part of the septum. A computer tomography (CT) scan and a biopsy suggestive of benign schwannoma were obtained before the lesion was removed by a degloving approach. The preoperative diagnosis of nasal septum schwannoma was confirmed. The patient is asymptomatic and without endoscopic evidence of recurrence 7 years after surgery. A review of the literature with particular emphasis on the clinical presentation, histological features, differential diagnosis and therapeutic options for such a rare lesion is included. PMID- 11052257 TI - Robert Feulgen Prize Lecture. Laser tweezers and multiphoton microscopes in life sciences. AB - Near infrared (NIR) laser microscopy enables optical micromanipulation, piconewton force determination, and sensitive fluorescence studies by laser tweezers. Otherwise, fluorescence images with high spatial and temporal resolution of living cells and tissues can be obtained via non-resonant fluorophore excitation with multiphoton NIR laser scanning microscopes. Furthermore, NIR femtosecond laser pulses at TW/cm2 intensities can be used to realize non-invasive contact-free surgery of nanometer-sized structures within living cells and tissues. Applications of these novel versatile NIR laser-based tools for the determination of motility forces, coenzyme and chlorophyll imaging, three-dimensional multigene detection, non-invasive optical sectioning of tissues ("optical biopsy"), functional protein imaging, and nanosurgery of chromosomes are described. PMID- 11052258 TI - Cell cycle maintenance and biogenesis of the Golgi complex. AB - How organelle identity is established and maintained, and how organelles divide and partition between daughter cells, are central questions of organelle biology. For the membrane-bound organelles of the secretory and endocytic pathways [including the endoplasmic reticulum (ER), Golgi complex, lysosomes, and endosomes], answering these questions has proved difficult because these organelles undergo continuous exchange of material. As a result, many "resident" proteins are not localized to a single site, organelle boundaries overlap, and when interorganellar membrane flow is interrupted, organelle structure is altered. The existence and identity of these organelles, therefore, appears to be a product of the dynamic processes of membrane trafficking and sorting. This is particularly true for the Golgi complex, which resides and functions at the crossroads of the secretory pathway. The Golgi receives newly synthesized proteins from the ER, covalently modifies them, and then distributes them to various final destinations within the cell. In addition, the Golgi recycles selected components back to the ER. These activities result from the Golgi's distinctive membranes, which are organized as polarized stacks (cis to trans) of flattened cisternae surrounded by tubules and vesicles. Golgi membranes are highly dynamic despite their characteristic organization and morphology, undergoing rapid disassembly and reassembly during mitosis and in response to perturbations in membrane trafficking pathways. How Golgi membranes fragment and disperse under these conditions is only beginning to be clarified, but is central to understanding the mechanism(s) underlying Golgi identity and biogenesis. Recent work, discussed in this review, suggests that membrane recycling pathways operating between the Golgi and ER play an indispensable role in Golgi maintenance and biogenesis, with the Golgi dispersing and reforming through the intermediary of the ER both in mitosis and in interphase when membrane cycling pathways are disrupted. PMID- 11052259 TI - Development of hormonal peptides and processing enzymes in the embryonic avian pancreas with special reference to co-localisation. AB - Studies on the developing mammalian pancreas have suggested that insulin and glucagon co-exist in a transient cell population and that peptide YY (PYY) marks the earliest developing endocrine cells. We have investigated this in the embryonic avian pancreas, which is characterised by anatomical separation of insulin and glucagon islets. Moreover, we have compared the development of the endocrine cells to that of processing enzymes involved in pancreatic hormone biosynthesis. PYY-like immunoreactivity occurred in islet cells from the youngest stages examined: it increased in amount from approximately 5 days of incubation and was co-localised with glucagon and to a lesser extent with insulin. Insulin and glucagon cells were numerous: co-existence of the two peptides in the same cells was but rarely observed. From the youngest stages examined, prohormone convertase (PC) 1/3-like immunoreactivity was detected in insulin cells and PC2-, 7B2- and carboxypeptidase E-like immunoreactivity in both glucagon and insulin cells. We conclude that: (1) PYY-like immunoreactivity occurs in avian islet cells but generally in lesser amounts than in mammals at the earlier stages, (2) the paucity of cells co-expressing insulin and glucagon indicate that all avian insulin cells do not pass through a stage where they co-express glucagon and (3) the early expression of the enzymes responsible for the processing of prohormones suggests that this process is initiated soon after islet cells first differentiate. PMID- 11052260 TI - Expression of fucosyltransferases in skin, conjunctiva, and cornea during human development. AB - During human development, type-1-precursor, sialyl-Le a, and Le x antigens were present in the periderm of skin and eye at week 6. The Le x antigen disappeared from cornea at 10 weeks and then from skin at 20 weeks. H-type-1, Le a, Le b, sialyl-Le a, H-type-2, sialyl-Le x, and Le y were found in cornea, conjunctiva, and periderm between 10 and 20 weeks. They disappear from the skin (at week 20) and progressively reappear in skin derivatives, especially in the epithelium of sweat glands. The secretory part of the sweat gland is type-1-precursor and H type-1 positive while its excretory part is Le a, Le b, sialyl-Le a, and Le y positive. On the eye surface the disappearance of Le x at 10 weeks and of the H type-1, sialyl-Le x, and Le y at week 35 starts in the central cornea in front of the lens. The corneal epithelium and the conjunctiva have similar antigens to those of excretory and secretory parts of the sweat gland, respectively. Invaginations and folding of the epidermis might preserve the embryonic staining. We propose that fucosylation patterns are associated with the embryonic origin and differentiation stage of tissue. The early and transient presence of Le x is associated with FUT4 or FUT9 activities, while the late appearance of Lewis antigens is related to other alpha3-fucosyltransferases. PMID- 11052261 TI - Vertebrate phylogeny of antigen D10: identification of a conserved foregut cell lineage. AB - The monoclonal antibody 5HL-5D11-D10 to antigen D10 identifies a cell lineage that is restricted to certain tissues of the human foregut. We investigated the tissue distribution of antigen D10 in mammals, birds, reptiles, amphibians and fish by immunohistochemical staining. Tissue from human and each of ten other mammalian species showed staining of gastric mucous neck cells and glands of the cardia and antrum, Brunner's glands, peribiliary glands and periductal glands of the pancreas. Six of the mammalian species also expressed antigen D10 in mucosa of the larger bronchi, and five expressed it to varying degree in small bowel distal to the duodenum and in colon (three of these five species). Antigen was not detected in any of the three species of bird studied. Both reptiles and amphibians showed strong staining for antigen D10 in the gastric mucous neck cells and pyloric glands, and in a subpopulation of secretory cells in the oesophagus, with the amphibian also expressing antigen in some epithelial cells of the mouth and lung. Although absent from two species of bony fish, antigen D10 was expressed by small groups of epithelial cells of the intestine of a shark, and generally by the epithelial and connective tissue cells of the gut and gills, and hepatocytes of one species of ray. The presence of antigen D10 in different tissues and species was confirmed by both an indirect ELISA and immunoblot analysis of tissue extracts. Our observations suggest that the D10 epitope characterises a subpopulation of mucus-secreting cells, predominantly of the foregut and associated organs, which has been conserved throughout terrestrial vertebrate evolution. PMID- 11052262 TI - Immunolocalisation of BPTI-like serine proteinase inhibitory proteins in mast cells, chondrocytes and intervertebral disc fibrochondrocytes of ovine and bovine connective tissues. An immunohistochemical and biochemical study. AB - A polyclonal anti-bovine pancreatic trypsin inhibitor (BPTI) IgY was raised in chickens immunised with aprotinin. The anti-BPTI IgY was subsequently isolated from egg yolks and purified to homogeneity by affinity chromatography on immobilised aprotinin and by Superose 6 size exclusion fast protein liquid chromatography (FPLC). Immunoblotting with the chicken IgY demonstrated its specificity for BPTI; 3.9 ng BPTI could be detected by this technique. There was no crossreactivity against alpha1-proteinase inhibitor (human and sheep), inter alpha-trypsin inhibitor (human and sheep), secretory leucocyte proteinase inhibitor or a range of serine proteinase inhibitory proteins (SPIs) isolated from plant sources (soybean and lima bean trypsin inhibitor, potato trypsin and chymotrypsin inhibitors) or serum SPIs (antithrombin-III, alpha2-macroglobulin). Immunoblotting using the anti-BPTI IgY identified the 6- to 12- and 58-kDa forms of endogenous ovine cartilage SPIs in cartilage extracts, confirming the interrelationship of the ovine cartilage SPIs with BPTI. BPTI-domain SPIs were immunolocalised within mast cells of ovine and bovine duodenum, lung and pancreas, and in ovine and bovine bronchial cartilage chondrocytes, chondrocytes of the superficial and intermediate zones of articular cartilage and in the fibrochondrocytes/chondrocytes of the nucleus PMID- 11052263 TI - The catenin, p120ctn, is a common membrane-associated protein in various epithelial and non-epithelial cells and tissues. AB - The cadherin-binding catenin p120ctn was originally identified as an Src-tyrosine kinase substrate. More recently, p120ctn has been shown in some cell types to be associated with catenin/cadherin complexes of adherens junctions. To address the question whether p120ctn is restricted to certain cell types or whether it is a general cellular component we investigated tissue distribution of p120ctn by immunohistochemistry and immunoblotting in the rat. We found p120ctn to be widely distributed in several tissues where it is mainly restricted to the plasma membrane. In various epithelia p120ctn was found in association with different adherens junctions such as the zonula adherens and puncta adherentia. In addition, p120ctn was localized along infoldings of the basal cell membrane, most prominently in renal proximal and distal tubules. pl20ctn was not restricted to epithelia. It was also found at intercalated discs of cardiomyocytes. In the nervous system, immunostaining was particularly prominent in areas rich in synapses suggesting that pl20ctn is a component of synaptic adherens junctions as well. By immunoblotting, four different isoforms of pl20ctn could be detected displaying similar electrophoretic mobilities as the isoforms 1A, 1B, 2A, and 2B reported from mice. Whereas all epithelia assayed contained at least two isoforms, testis, heart, brain, and retina contained a single 110-kDa band that corresponds to isoform 1B in mice. PMID- 11052264 TI - Temporospatial expression of tissue inhibitors of matrix metalloproteinases-1, -2 and -3 during development, growth and aging of the mouse skeleton. AB - Proteolytic degradation of collagen-rich extracellular matrices is a key feature in the development, growth and aging of skeleton. Matrix metalloproteinases (MMPs) are a family of enzymes capable of performing this function, whereas tissue inhibitors of MMPs (TIMPs) are believed to play an important role in regulating their activity. To better understand the roles of TIMP-1, -2 and -3, we have studied their mRNA levels in several different mouse tissues with special emphasis on the skeleton and the developing eye. A systematic analysis of TIMP-1, -2 and -3 mRNA levels in mouse knee joints during growth and aging demonstrated markedly different expression patterns for each TIMP. Immunohistochemical analysis revealed several time-dependent changes in the distribution of TIMP-1 and -2 in articular and growth cartilages, synovial tissue and bone. The data suggest that upon aging synovial tissue becomes the major source of synovial fluid TIMPs. In articular cartilage these inhibitors were mainly found in the deep layer and in subchondral bone. Compared with epiphyseal growth plate, the amounts of TIMP-1 and -2 in articular cartilage were quite low. These findings suggest that the capacity of articular cartilage chondrocytes to inhibit MMP activities by local production of TIMPs is limited, which may be of consequence during osteoarthritic cartilage degeneration. PMID- 11052265 TI - Lipids are a constitutive component of cytolytic granules. AB - Cytolytic granules are specific organelles of activated cytotoxic lymphocytes mediating storage and regulated excretion of lytic molecules for killing of target cells. A variety of the other granule components may also participate in granule-mediated cytotoxicity. In this study, the subcellular localization of lipids in the granules of human decidual CD56+ natural killer-like cells was determined by staining with malachite green aldehyde and imidazole-buffered osmium tetroxide. Lipids were shown, for the first time, to be a constitutive component of cytolytic granules. Lipids formed an additional structural microdomain, located between the granule-limiting membrane and the granule core. Images of the granules on serial sections suggested that intragranular lipids wrap the core. We speculate that granule lipids participate in packing of lytic molecules inside the granules, in autocrine signaling ending granule secretion, and in the killing process. PMID- 11052266 TI - Neurotoxicity of calcineurin inhibitors: impact and clinical management. AB - Between 10%-28% of patients who receive the immunosuppressant cyclosporine (CsA) experience some form of neurotoxic adverse event. Both sensorial motoric functions may be adversely affected, and thus patients present with a wide range of neurological and psychiatrical disorders. Mild symptoms are common and include tremor, neuralgia, and peripheral neuropathy. Severe symptoms affect up to 5 % of patients and include psychoses, hallucinations, blindness, seizures, cerebellar ataxia, motoric weakness, or leukoencephalopathy. Tacrolimus is associated with similar neurotoxic adverse events. Neurotoxicity may result in serious complications for some patients, particularly recipients of orthotopic liver transplants. Factors that may promote the development of serious complications include advanced liver failure, hypertension, hypocholesterolemia, elevated CsA or tacrolimus blood levels, hypomagnesemia, and methylprednisolone. Occipital white matter appears to be uniquely susceptible to the neurotoxic effects of CsA; injury to both the major and minor vasculature may cause hypoperfusion or ischemia and local secondary toxicity in the white matter. Calcineurin inhibition by CsA and tacrolimus alters sympathetic outflow, which may play a role in the mediation of neurotoxic and hypertensive adverse events. The symptoms of CsA- and tacrolimus-associated neurotoxicity may be reversed in most patients by substantially reducing the dosage of immunosuppressant or discontinuing these drugs. However, some patients have experienced permanent or even fatal neurological damage even after dose reduction or discontinuation. CsA-sparing and tacroli-mus-sparing drug regimens that use the immunosuppressant mycophenolate mofetil, which has no neurotoxic effects, may reduce the incidence and severity of neurotoxic adverse events while maintaining an adequate level of immunoisuppression. PMID- 11052267 TI - Noninvasive assessment of cardiac risk in type I diabetic patients being evaluated for combined pancreas-kidney transplantation using dipyridamole-MIBI perfusion tomographic scintigraphy. AB - This study was performed to determine the value of dipyridamole-99m Tc-methoxy isobutyl isonitrile perfusion (99mTC-MIBI) tomographic scintigraphy in the assessment of cardiac risk in patients being evaluated prior to combined pancreas kidney transplantation (PKT). We performed perfusion tomographic scintigraphy using single photon emission computed tomography (SPECT) on 77 patients. The tomographic images did not show clinically relevant findings in 65 patients. In the remaining 12 patients, coronary arteriography was performed: 2 showed normal results, 4 showed no stenosis, and 6 showed significant stenosis ( > or = 70%). Seventy-two patients underwent PKT. During the follow-up (6-48 months), there were seven cardiac events, 4 patients with significant stenosis, and 3 with nonsignificant stenosis upon coronary arteriography, and all had pathological tomographic images. 99mTc-MIBI tomographic scintigraphy may be useful in identifying patients at low risk of incurring cardiac events after PKT and may, in a large group of patients, obviate the need for routine coronary angiography. PMID- 11052268 TI - Hepatic grafts from live donors: donor morbidity for 470 cases of live donation. AB - Living donor-morbidity was evaluated in 470 consecutive cases of living donor liver transplantation carried out from June 1990 to May 1999 at Kyoto University. Grafting was categorized into 4 groups according to the resection lines; left lateral segmentectomy (S2 + 3, n = 282, R1), extended left lateral segmentectomy without middle hepatic vein (MHV) (S2 + 3 + part4, n = 45, R2), left lobectomy with MHV (S2 + 3 + 4, n = 99, R3) and right lobectomy without MHV (S5 + 6 + 7 + 8, n = 43, R4). Intraoperative blood loss and operation duration were less for left lateral segmentectomy, but no significant difference was observed between left lobectomy and right lobectomy. The length of postoperative hospital stays was comparable among all groups except for the group with right lobe grafting. The AST values at the peak and at POD 7 were significantly elevated for right lobectomy, but the AST value normalized within one month in the majority of the cases. The close follow-up of donors with more than 1,000 ml intraoperative bleeding, and of those donors who stayed in hospital for more than 30 days, the close follow-up, furthermore, of those donors with AST values higher than 100 IU/ L AST after one month, revealed complete recovery. Biliary leakage was the most common and annoying complication after donor operations, especially in for right lobe grafting, but all donors recovered completely with conservative or minimal invasive therapy. The two cases of re-operation due to adhesive mechanical ileus we encountered were resolved completely. Finally, no donor-operation related death was noted. In conclusion, the morbidity of living donors is low or minimal even for right lobectomy, the most extended procedure, and complete recovery can be expected in all cases. PMID- 11052269 TI - Native pyeloureterostomy after kidney transplantation: experience in 48 cases. AB - Necrosis and stenosis of the ureter are severe complications after kidney transplantation and occur with mean incidence of 2,9-13,4 %. Several surgical techniques like simple nephrostomy or complex urinary tract reconstruction have been applied for repair. In this study, our experience with native pyeloureterostomy (NPUS) using the native ureter is presented. Between March 1978 and June 1996, 2,592 kidney transplantations were performed in our institution. In 48 patients (1,9%), secondary urinary tract reconstruction by NPUS was necessary. These patients were evaluated retrospectively by review of the case notes. At the time of operation the mean age was 45 +/- 14 years. Indications for NPUS were distal ureteral stenosis (n = 29), necrosis (n = 17), bleeding (n = 1) or iatrogenic lesion of the ureter (n = 1). The mean time period between transplantation and urinary tract reconstruction was 20 +/- 23 days (range: 1-90 days) for necrosis and 404 +/- 637 days (range: 14-2,385 days) for stenosis. A pyeloureterostomy was technically feasible in all patients using the recipient's ipsilateral ureter. In 40 out of 48 patients the graft developed a normal function postoperatively (follow up: 39 +/- 48 months). A graft nephrectomy was necessary only in one patient, because of complete pyelonnecrosis 6 days after NPUS. Two grafts were lost due to acute rejection. Data of five patients were not available > 15 years after successful reconstruction. We can conclude that NPUS is a safe and simple rescue technique for the treatment of distal ureteral complications after kidney transplantation. Therefore, this technique should be the therapy of choice when secondary reconstruction by re-ureteroneocystostomy is not possible. PMID- 11052270 TI - Completely reversed acute rejection is not a significant risk factor for the development of chronic rejection in renal allograft recipients. AB - Although acute rejection (AR) has been shown to correlate with decreased long term renal allograft survival, we have noted AR in recipients who subsequently had stable function for more than 5 years. We reviewed 109 renal graft recipients with a minimum of 1 year graft survival and follow-up of 5-8 years. Post transplant sodium iothalamate clearances (IoCI) measured at 3 months and yearly thereafter were used to separate recipients into 2 groups. In 61 patients (stable group), there was no significant decrease ( > 20 % reduction in IoCl over 2 consecutive years) in IoCl. Forty-eight patients had significant declines in IoCl (decline group). Groups were compared for incidence, severity, timing, and completeness of reversal of AR. Rejection was considered completely reversed if the post-AR serum creatinine (Scr) returned to or below the pre-AR nadir Scr after anti-rejection therapy. The incidence of AR was not significantly different between groups (47% vs 52%). A trend toward a lower mean number of AR episodes per patient was noted in the stable group (0.69 vs 1.04, P = 0.096), but the timing of AR was not different. Steroid-resistant AR occurred in approximately 25 % of both groups. A striking difference was seen in complete reversal of AR, with the stable group having 100% (42/42 episodes of AR in 29 patients) complete reversal whereas only 32 % (8/25) of the patients in the decline group had complete reversal (P < < 0.001). Of 8 declining patients with complete reversal, graft loss was due to chronic rejection (CR) in only 3. Seventeen declining patients had incomplete reversal of AR, and 82 % (14/17) lost their grafts to CR. Overall, only 8% (3/37) of the recipients with complete reversal of AR developed CR. No patients with incompletely reversed AR had stable long-term function as measured by IoCl. AR is not invariably deleterious to long-term renal graft function if each episode of AR can be completely reversed. PMID- 11052271 TI - Attitudes of hospital staff involved in organ donation to the procedure. AB - Hospital staff have a key function in asking for potential organ donors, but little is known about their own attitudes towards donation. In a community hospital with 7-8 multi-organ extraction procedures each year 199 staff members were surveyed. Although only 7% of the responding staff would personally refuse to donate an organ, 23 % would not give consent to organ donation from a close relative. 47 % of those prepared to be donors had signed a donor card. Donors informed their family more frequently (88 %) about their personal attitude towards organ donation than non-donors (60 %), or undecided personnel (43,8 %; chi-square P = 0,004). No significant difference in attitude according to medical profession subgroups was found. The findings are in line with general population surveys and indicate that much work needs to be done to encourage medical staff involved in organ donation to set an example to the community. PMID- 11052272 TI - Plasma levels of endothelin-1 in patients with the hepatorenal syndrome after successful liver transplantation. AB - The hepatorenal syndrome (HRS) is characterized by renal vasoconstriction leading to deterioration of renal function in patients with liver disease. A possible role of endothelin-1 (ET-1) in the pathogenesis of HRS has been suggested, but a correlation between ET-1 plasma levels and the development of HRS as well as the recovery from HRS following OLT has not been shown yet. We performed longitudinal measurements of ET-1 plasma levels in four groups of patients, 5 patients with HRS before and after orthotopic liver transplantation (OLT), 10 patients without HRS undergoing OLT, 20 patients with chronic renal failure but without liver disease, and 12 healthy controls. Before OLT, plasma levels of ET-1 were higher in patients with HRS (19.5 +/- 8.6 ng/l, P < 0.001; n = 5) compared to patients without HRS (4.9 +/- 1.1 ng/l; n = 10), normals (1.2 +/- 0.18 ng/l; n = 12), and patients with chronic renal failure (2.4 +/- 0.4 ng/l; n = 20). Patients with HRS compared to patients without HRS had higher levels for creatinine (2.42 +/- 0.6 vs. 0.89 +/- 0.05 mg/dl, P < 0.05), creatinine clearance (107 +/- 9 ml/min vs. 44.6 +/- 5.5 ml/ min, P < 0.001), and bilirubin (11.4 +/- 3.8 vs. 3.7 +/- 1 mg/dl, P < 0.05) before OLT. Within one week after OLT, there was a rapid decrease in ET-1 levels in patients with HRS while creatinine and bilirubin levels decreased slower. Regression analysis revealed a weak correlation between serum creatinine and ET-1 (r = 0.192, P = 0.04) and a significant correlation between serum bilirubin and ET-1 (r = 0.395, P < 0.001). The means of the ET-1 levels decreases rapidly with improvement of liver function after OLT. Levels of ET-1 correlate with excretory liver function assessed by bilirubin. The fall in ET-1 levels preceding improvement of renal function further strengthens the concept of ET-1 being a causative factor in HRS. PMID- 11052273 TI - Complement activation by anti-endothelial cell antibodies in MHC-mismatched and MHC-matched heart allograft rejection: anti-MHC-, but not anti non-MHC alloantibodies are effective in complement activation. AB - Classical complement activation is a major effector mechanism in the development of vascular lesions contributing to allograft rejection. We investigated complement activation by alloantibodies reactive with graft endothelial cell (EC) alloantigens in settings of MHC-mismatched and MHC-matched (non-MHC-mismatched) rat heart transplantation (Tx). Allosera and heart allografts were collected at the day of rejection (day 7-8 and day 28-35 in MHC-mismatched and non-MHC mismatched Tx respectively) or earlier. Allosera reactivity was studied in vitro using rat-heart-endothelial-cell (RHEC) lines expressing the appropriate donor MHC and non-MHC alloantigen profile. Immunohistochemical analysis of rejected heart allografts showed deposition of alloantibodies in both MHC-mismatched and MHC-matched heart allografts, but expression of C3 was only seen in the vasculature of MHC-mismatched grafts. FACS analysis showed that anti MHC as well as anti non-MHC allosera were reactive with donor EC cell surface antigens. Both sera had similar IgG subclass profiles of anti-endothelial cell antibodies. Complement activation by anti MHC and anti non-MHC alloantibodies on EC was measured by FACS analysis of C3 and C5b-9 (MAC) expression. Distinct expression of C3 was noticed for EC incubated with anti-MHC allosera, but hardly for EC incubated with anti non-MHC allosera. C5b-9 was low but showed no difference between the two allosera. However, complement-mediated cytotoxicity experiments showed that functional (lytic) MAC was induced with anti MHC allosera but hardly with anti non-MHC allosera. Our data show that in settings of MHC-matched heart transplantation alloantibodies against endothelial non-MHC alloantigens are generated, but, in contrast to alloantibodies to MHC alloantigens, these alloantibodies have only poor complement-activating and lytic potentials. Whether anti non-MHC allolantibodies effect other biological processes relevant for heart allograft vasculopathy, including development of graft arteriosclerosis, needs further elucidation. PMID- 11052274 TI - Arterial complications after liver transplantation. AB - From September 1988 through April 1998, 1,000 liver transplantations were performed on 911 patients. During the postoperative control examinations of 837 patients, we found 23 (2.74 %) with hepatic artery thromboses, 27 stenoses of the hepatic artery (3.22 %), and 6 aneurysms of the graft artery. Seventeen patients underwent retransplantation because of arterial complications. Depending on the clinical symptoms, we treated both the local situation as well as the resulting complications of inadequate arterial graft flow. The aneurysms were primarily treated surgically. The first choice of treatment of stenoses was balloon angioplasty. Early postoperative artery thromboses were also treated surgically by thrombectomy in selected cases. For the resulting biliary and local septic complications we preferred endoscopic and drainage procedures. Our clinical experiences have led us to find pretransplantation angiography recommendable, especially in the case of splanchnic artery stenoses, for bypassing from the aorta for arterial perfusion of the graft. PMID- 11052275 TI - Diagnosis of cytomegalovirus retinitis after heart transplantation. PMID- 11052276 TI - Early detection of an ureterovesical urinary leakage in a patient with normal renal function following kidney transplantation. PMID- 11052277 TI - Placement of interatrial patch suture lines in atrioventricular canal defect repair. AB - BACKGROUND: The placement of the suture line for interatrial patches in complete and incomplete atrioventricular canal defect repairs varies from surgeon to surgeon despite established anatomic knowledge of the atrioventricular conduction system. This study describes our technique for it and reviews early and long-term outcomes. METHODS: Between 1980 and 1999, 64 infants and children underwent repair of either complete (n=39) or incomplete (n=25) atrioventricular canal defects. Thirty-four of the children (53.1%) had Down's syndrome. The suture line for the interatrial patch originated on either the artificial or native ventricular septal crest and continued leftward above the annulus of the left inferior leaflet of the atrioventricular valve at the posteroinferior corner. All stitches were placed in a horizontal mattress or U-shaped fashion. RESULTS: The operative survival rate was 94% (4 early deaths) and the overall survival rate was 85% (6 late deaths). Atrioventricular heart blocks occurred in none of the patients. Although left-sided atrioventricular function significantly improved with repair, two patients (3.1%) required reoperation for valve replacement because of residual or recurrent insufficiency. CONCLUSIONS: This suture technique for interatrial patches is straightforward and results in a low incidence of heart block and a low re-operation rate for left atrioventricular valve insufficiency. PMID- 11052278 TI - Neurogenic vasoreactivity of human internal thoracic artery is unaffected by endothelial cell integrity. AB - BACKGROUND: The role of internal thoracic artery (ITA) nervous supply has not been previously considered as a potential factor influencing excellent long-term patency of an ITA graft. To define the interaction between the primary afferent neurons and endothelial cells of ITA, we investigated the effects of acute capsaicin administration in vitro on the isometric tension of human ITAs. METHODS: Vessels were obtained from patients undergoing coronary bypass or from multi-organ transplant donors. Thirty-three ITA segments (5 mm wide) were suspended as rings between two stainless-steel stir-ups in water-jacketed (37 degrees C) tissue baths. The tissue baths contained 10 ml physiological salt solution (PSS) of the following composition (mM/L): NaCl 119, KCl 4.7, NaH2PO4 1.0, MgCl2 0.5, CaCl2 2.5, NaHCO3 25, and glucose 11, aerated continuously with 95% O2 and 5% CO2. Peptidase inhibitors, phosphoramidon (1 microM) and captopril (1 microM), were added to PSS to decrease peptide degradation. Mechanical responses were measured isometrically and recorded on a polygraph via isometric force transducers. Vessels were preconstricted with submaximal concentrations of norepinephrine. After the tension had stabilized, capsaicin was added cumulatively to the tissue bath. The viability of ITA was verified by its responses to endothelial-dependent (acetylcholine, 1 microM) (n=20) and endothelial-independent (sodium nitroprusside, 10 microM) (n=13) vasodilators. RESULTS: The exposure of capsaicin (3 microM) to human ITA produced varied effects on ITA irrespective of its endothelium. Capsaicin induced contraction of the ITA smooth muscle in 13 endothelium-intact ITA segments while it produced vasoconstriction in 9 endothelium-denuded ITAs (p=0.6437). In response to capsaicin, relaxation of ITA smooth muscle was observed in 7 ITA rings with endothelium, while vasodilation was present in 4 ITA segments without endothelium (p=0.4099). CONCLUSIONS: Capsaicin-sensitive neurons encircling human ITA produce a neurogenic vasoreactive response independent of ITA endothelial cell integrity. PMID- 11052279 TI - Effect of ATP-potassium channel opener nicorandil on long-term cardiac preservation. AB - BACKGROUND: ATP-sensitive potassium channels have been shown to be one of the important protective mechanisms for the ischemic myocardium. The purpose of this study was to evaluate the protective effect of nicorandil, an ATP-sensitive potassium channel opener, on myocardium during 6 hours hypothermic preservation. METHODS: Preserved rat hearts were randomly divided into 4 groups according to cardioplegia and preservation protocols as follows: (1) histidine-tryptophan ketoglutarate solution (HTK) for both cardioplegic and immersing solutions (group A); (2) nicorandil-added HTK for cardioplegic solution and nicorandil-free HTK for immersing solution (group B); (3) nicorandil-free HTK for cardioplegic solution and nicorandil-added HTK for immersing solution (group C); and (4) nicorandil-added HTK for both cardioplegic and immersing solutions (group D). RESULTS: The recovery of postischemic cardiac function, including left ventricular developed pressure and end-diastolic pressure, was significantly improved in group B and group C as compared with the other groups (p<0.05). Postischemic intracellular calcium concentration was significantly lower in group B and group C than in group A (p<0.05). CONCLUSIONS: We concluded that nicorandil induced hyperpolarizing arrest could reduce ischemia-derived myocyte injury and inhibit the influx of calcium into the myocytes in long-term cardiac preservation. PMID- 11052280 TI - Myocardial fixation of anticardiac troponin I antibody and cardiac troponin I release. AB - BACKGROUND: The threefold aim of this experimental study was to test the correlation of cardiac troponin I released to myocardial infarction size and myocardial fixation of anticardiac troponin I antibody and to determine how long after myocardial infarction the measure of cardiac troponin I concentration can evaluate myocardial infarction size. METHODS: Forty rabbits were assigned either to a control group or to an experimental preconditioned group. Infarction was obtained by tightening a snare around the left anterior descending artery. Serial venous blood samples were drawn for measurement of cardiac troponin I. The rabbits were sacrificed at 72 hours and a histological study was performed to determine the infarct size and the size of the area void of fixation of anticardiac troponin I antibody. RESULTS: There was a linear correlation between the total amount of CTn I released and both infarct size (r=0.45, p<0.02) and the size of the area void of anti-cardiac troponin I antibody (r=0.47, p<0.02). These two sizes were strongly correlated (r=0.95, p<0.02). The hour 9 CTn I sample was the best correlated with both the infarct size (r=0.47, p<0.02) and the size of area void of anticardiac troponin I antibody (r=0.45, p<0.02). CONCLUSIONS: Our study shows that: 1) cardiac troponin I release is correlated to both myocardial infarction size and the size of area void of fixation of anticardiac troponin I antibody, 2) the area void of anticardiac troponin I antibody fixation includes the whole ischemic area, and 3) evaluation of myocardial infarction size can be obtained by CTn I concentration as early as the ninth hour. PMID- 11052281 TI - Assessment of perioperative predictive factors influencing survival in patients with postinfarction ventricular septal perforation. Classified by the site of myocardial infarction. AB - BACKGROUND: The present study was designed to identify the perioperative factors and to consider a counterplan for the improvement of surgical results, based on the site of myocardial infarction. METHODS: Sixteen patients with postinfarction ventricular septal perforation underwent surgical repair. The operation was performed 5+/-3 days after the onset of ventricular septal perforation using the same method, an infarctectomy and reconstruction of the septum and right and left ventricular walls with a single Dacron patch. The ventricular septal perforation was anterior in 11 patients and posterior in 5. Preoperative hemodynamics between survivors and non-survivors were compared. Left ventricular wall motion was estimated using echocardiography by wall motion score (divided into 17 segments and each segment was graded on a fourpoint scale: normal, 0; hypokinetic, 1; severe hypokinetic, 2; a- or dyskinetic, 3) and summed up. RESULTS: The operative mortality was 36% in 11 patients with anterior ventricular septal perforation. In non-survivors compared to survivors, wall motion score was greater (25+/-4 vs 18+/-4, p<0.01) and all values were over 20. The value of the cardiac index divided by Qp/Qs was lower (0.98+/-0.09 vs 1.44+/-0.31, p<0.02) and all were under 1.1. In 5 patients with inferior ventricular septal perforation, the operative mortality was 40%. In non-survivors compared to survivors, wall motion score was greater (18, 18 vs 7, 2, 12) and the right atrial pressure was greater (18, 19 vs 10, 9, 9 mmHg) due to a right ventricular infarction. CONCLUSIONS: The patients with poor left ventricular wall motion were lost for reasons unrelated to the site of myocardial infarction. Moreover, a cardiac index over Qp/Qs in anterior ventricular septal perforation and the existence of a right ventricular infarction in inferior ventricular septal perforation was predictive of operative mortality. PMID- 11052282 TI - Collagen patch coated with fibrin glue components. Treatment of suture hole bleedings in vascular reconstruction. AB - BACKGROUND: Bleeding from suture holes during vascular reconstruction, particularly when polytetrafluoroethylene (PTFE) prostheses are used, is still a problem which can lead to intraoperative delay and increased blood loss. The aim of this prospective, randomised, open, controlled multicentre study was to evaluate whether the use of a new local haemostyptic would reduce intraoperative blood loss and the time to haemostasis. METHODS: Thirty patients received a new haemostyptic (TachoComb H, Nycomed Pharma AG), whereas another 30 patients were treated with compresses. The vascular reconstructions were either anastomoses or patch angioplasties and were performed using PTFE vascular prostheses. RESULTS: The mean time to haemostasis of suture hole bleeding in the haemostyptic group (326.0 sec) was significantly shorter compared to the control group (514.3 sec) (p=0.006). The median intraoperative blood loss was 24.5 g in the treatment group and 57.3 g in the control group (p=0.045). CONCLUSIONS: It was shown that collagen patches coated with components of fibrin glue significantly reduce the time to haemostasis as well as blood loss at the operation site in patients undergoing vascular reconstruction with PTFE grafts. PMID- 11052283 TI - Evaluation of body fluid status after cardiac surgery using bioelectrical impedance analysis. AB - BACKGROUND: In the assessment of fluid status after cardiac surgery, we applied bioelectrical impedance analysis (BIA) to measure the total body water (TBW), extracellular fluid (ECF), and intracellular fluid (ICF), and evaluated its validity. METHODS: Thirty patients who underwent cardiopulmonary bypass (CPB group) and 19 surgical patients not receiving CPB (non-CPB group). RESULTS: The change of BIA values (deltaTBW, deltaECF, deltaICF), body weight and cumulative fluid balance were determined for 120 postoperative hours, and the relationship between BIA values and body weight and fluid balance were evaluated. Postoperative changes in BIA values in the CPB group were compared with those in the non-CPB group. Finally the ECF/ICF ratio and hemodynamic parameters were compared. deltaTBW and deltaECF correlated with changes in body weight and fluid balance, respectively. Especially there was a high correlation in each case although large deviations in the slope of the regression lines were observed. TBW and ECF increased from immediately after operation up to 96 hours (the maximum value was at day 2). On the other hand, ICF decreased from 48 to 72 hours after operation. There were significant high ECF/ICF in the CPB group compared with the non-CPB group from 12 to 72 postoperative hours. We found that ECF/ICF correlated inversely with mean blood pressure, mixed venous oxygen saturation and colloid osmotic pressure, and positively with central venous pressure and pulmonary artery wedge pressure. CONCLUSIONS: It was considered that BIA was useful for evaluating the relative changes in TBW and fluid distribution, and ECF/ICF might be a new parameter for abnormal water metabolism after cardiac surgery. PMID- 11052284 TI - Resolution of protein-losing enteropathy with standard high molecular heparin and urokinase after Fontan repair in a patient with tricuspid atresia. AB - At 6 years of age, a girl with tricuspid atresia underwent a Bjork modified Fontan procedure with implantation of a Carpentier Edwards bioprosthesis between the right atrium and the right ventricle. Ten years later she developed increasing edema, ascites and pleural effusions. The work-up showed severe stenosis of the bioprosthesis and protein losing enteropathy with a massive decrease of the albumin level to 14 g/l (normal 40-50 g/l). At 17 years of age, the bioprosthesis was replaced with a direct anastomosis between the cavoatrial junction and the right pulmonary artery. Within one month post-operatively, extensive thrombosis of the superior vena cava, anonymous and subclavian veins occurred. Protein-losing enteropathy persisted with an albumin level of 17 g/l. Parallel to the successful treatment of these thrombi with high molecular heparin and urokinase, protein losing enteropathy and hypoalbuminemia resolved completely as long as the antithrombotic treatment with high molecular heparin was continued. Oral anticoagulation was ineffective. Chronic antithrombotic treatment with high molecular heparin may thus be the treatment of choice in these forms of protein-losing enteropathy associated with venous thrombosis. PMID- 11052285 TI - Surgical management of echinococcosis of the heart. AB - A 23-year-old female with an echinococcal cyst in the atrioventricular groove of the heart is reported. The diagnosis and the location of the cyst were confirmed by echocardiography and cardiac catheter. Successful enucleating of the cyst with the aid of cardiopulmonary bypass and the length of follow-up is reported, along with a review of relevant literature. PMID- 11052286 TI - How to spread out the satisfactory exposure: placement technique in right-sided maze procedure. AB - We have, whenever possible, substituted cryoablation for incisions in the maze procedure. Cryoablation helps prevent massive bleeding because the number of resuturing sites is reduced and aortic cross-clamp time decreased. We are careful not to injure the atrio-ventricular conduction system during incision at the side of the Koch triangle or cryoablation while manipulating the atrial septum. In prior versions of the right-sided maze procedure, the IVC return cannula has obstructed the surgical field approaching the IVC and the tricuspid valve annulus. This report describes a new modification of maze procedure that includes changing the venous return site. We have obtained a satisfactory surgical field without being afraid of the anatomical pitfall, between the appendage and the tricuspid valve, and between the right coronary artery and the extended T incision. We studied 12 patients (8 males and 4 females, mean age 61 years) who underwent the maze procedure. Sinus rhythm was restored in 11 patients, whose clinical condition was improved to NYHA class I status. This technique has proven effective in obtaining satisfactory exposure of the-right atrium during the maze procedure. PMID- 11052287 TI - Monitoring aspects during port-access cardiac surgery. AB - OBJECTIVE: To report the experience gained at our Cardiosurgical Centre with the recently introduced port-access technique. METHODS: EXPERIMENTAL DESIGN: Prospective collection of data from the month of October 1997. SETTING: Regional University HospitaL Patients: Adult patients undergoing coronary bypass graft or mitral valve surgery. INTERVENTIONS: Port-access technique makes it possible to carry out open-heart procedures through a minithoracotomy and extrathoracic cardiopulmonary bypass with a set of properly designed catheters (Heartport EndoCPB system) for cardioplegia delivery and heart venting. MEASURES: Transesophageal echography and pressure traces are the main monitoring tools used for the correct placement of these catheters and for the clinical management of the patient. RESULTS: Sixty-two cases have been performed so far. A complete description of the procedure, with monitoring aspects and problems encountered is thoroughly presented. CONCLUSIONS: The major differences with traditional cardiac surgery are that interruption of myocardial perfusion is not achieved through a transversal clamp but through an endovascular occlusive balloon and that thoracic access is by minithoracotomy. Unlike traditional open surgery, the surgeon has no direct vision of the position of the clamp and the anesthesiologist can not visually inspect the contractile state of the heart. The operative team has to cope with a multifaceted system of monitored variables that must be continuously integrated and interpreted. Tight cooperation and continuous communication between anaesthesiologist, surgeons and perfusionist appear to be more important than in any other cardiac operation. PMID- 11052288 TI - Standards and concepts in valve surgery. PMID- 11052289 TI - An analysis of risk factors of perioperative bleeding in surgical repair of abdominal aortic aneurysm. AB - BACKGROUND: In surgical repair of abdominal aortic aneurysm (AAA), excessive bleeding which causes postoperative complications is sometimes observed. To determine the risk factors of perioperative excessive blood loss, this retrospective study was performed. METHODS: Design. A retrospective study. Setting. An academic medical center. Participants. One hundred and forty patients underwent elective surgical repair of an abdominal aortic aneurysm (AAA) at our institution from 1995 through 1997. Measurements. The present study includes critical review of 140 consecutive charts of patients undergoing elective surgical repair of AAA. Preoperative laboratory data, intraoperative data and amount of blood loss to identify risk factors of perioperative blood loss. Factors which were found to be significantly associated with the amount of perioperative blood loss were preoperative plasma fibrin degradation product (FDP) level (r=0.445), amount of immediate re-infusion of shed blood (r=0.438), and duration of operation (r=0.411). RESULTS: Preoperative fibrinogen level correlated with perioperative blood loss little (r=-0.187). Preoperative platelet count or the other coagulation profile did not affect the amount of perioperative blood loss. The patients whose preoperative FDP were more than 40 microg x ml(-1) significantly increased the risk of excessive blood loss compared with less than 40 microg x ml(-1). CONCLUSIONS: The significant preoperative risk factor of perioperative blood loss was only FDP level in present study. Especially, the patients whose preoperative FDP were more than 40 microg x ml(-1) increased the risk of excessive blood loss. PMID- 11052290 TI - Carotid surgery without angiography is possible and safe. AB - BACKGROUND: To evaluate the possibility to perform carotid surgery without angiography. METHODS: From January 1994 to June 1998, 514 patients with carotid obstructive disease were operated upon, 225 of them (43.8%) without previous angiography; 55 out of 68 (80.8%) during the last six months. Eighty-one (36.0%) had lateralizing symptoms, 50 aspecific ones (22.2%) and 94 were asymptomatic (41.8%). All patients were investigated by color-coded duplex sonography (CDS) of the arteries at the neck and by transcranial Doppler (TCD) and computed tomography (CT). One hundred eighty-eight patients were operated upon under local anaesthesia and 37 under general anesthesia; 204 had a carotid endartereotomy (90.7%) with patch angioplasty in 154 (75.5%), and 21 required a bypass graft (9.3%). In 26 patients (11.5%) an indwelling shunt was needed. RESULTS: Findings at surgery were consistent with CDS for plaque composition, ulcerations and degree of stenosis. There were no early deaths. Neurologic or ocular deficits occurred in 2 cases (0.9%). No strokes were observed in follow-up from 6 to 34 months. CONCLUSIONS: Carotid endarterectomy can be done without angiography in selected cases provided CDS plus TCD are of high quality. Under such conditions it can be considered a safer way to deal with carotid obstructive disease. PMID- 11052291 TI - Management of aberrant subclavian artery aneurysms. AB - An aberrant subclavian artery is the most common congenital anomaly of the aortic arch. Aneurysms of these vessels are relatively rare lesions. A high clinical suspicion must be maintained in patients with an abnormal mediastinum on chest X ray, especially in patients with dysphagia, dyspnea, or upper extremity ischemic symptoms not otherwise explained. Potentially disastrous complications, including spontaneous rupture and perforation into the esophagus may occur, and are invariably fatal. We present the case of an 1 asymptomatic 72-year-old male with an aberrant right c subclavian artery aneurysm presenting as a mediastinal mass on routine chest X-ray. Repair was by aneurysmectomy through a left thoracotomy and right common carotid artery to subclavian artery bypass via median sternotomy with resolution of his symptoms. We reviewed 74 cases in the English literature to February 1998. The pertinent anatomy is discussed and trends in surgical treatment are identified. Recent agreement on the surgical approach and choice of revascularization appears to exist, but advances in diagnostic and interventional radiologic capabilities have increased the number of asymptomatic lesions encountered and may alter the treatment of this lesion in the near future. PMID- 11052292 TI - Aortic dissection with intimal intussusception: diagnosis and management. AB - A man presented with acute chest and back pain with loss of consciousness. CT scan showed a mass in the arch that extended into the descending aorta. A diagnosis of type I aortic dissection was ultimately made by echocardiography. At surgery there was a circumferential intimal tear in the aortic root, and an intussuscepted dissection flap was retrieved from the arch and descending aorta. PMID- 11052293 TI - Chyloperitoneum. A rare complication after abdominal aortic aneurysm repair. AB - We report a case of chylous ascites as a rare complication following elective aortic aneurysm repair in a 66-year-old male. After its early development on the second post-operative day, re-laparotomy was performed with ligation of fistulas and omentumplasty. After recurrence of chylous ascites, conservative treatment for three months including parenteral nutrition and low-fat diet under continuous peritoneal drainage led finally to success. PMID- 11052294 TI - Concomitant abdominal aortic aneurysm repair and cholecystectomy. Combination of retroperitoneal aortic reconstruction and gassless laparoscopic cholecystectomy. AB - The coexistence of cholelithiasis and abdominal aortic aneurysm is not uncommon. However, cholecystectomy at the time of abdominal aortic reconstruction has generally been delayed because of the potential contamination of the graft. The case described here had concomitant cholelithiasis and abdominal aortic aneurysm, both of which were required to be treated, and was successfully treated with a combination of retroperitoneal abdominal aortic reconstruction and gasless laparoscopic cholecystectomy. PMID- 11052295 TI - Aneurysm dissection of the extracranial internal carotid artery. AB - Aneurysm of the extracranial internal carotid artery is a rare event. This is a pathology with an elevated mortality of 70%. The most important etiologic factor is atherosclerosis. Here we report a case of surgically treated extracranial internal carotid artery (ICA) aneurysm. A 77-year-old man noticed a laterocervical pulsatile mass. Color Doppler ultrasonography revealed an ICA aneurysm related to a parietal thrombosis. The ICA aneurysm was confirmed by intra-arterial digital subtraction angiography, and cerebral computer tomography (CT) was negative. Surgical treatment reconstruction with the interposition of a part of the great saphenous vein was indicated. Exploration of the aneurysmatic wall revealed a posterolateral dissection. In this paper are discussed clinical and therapeutic implications. PMID- 11052296 TI - Critical limb ischemia in a patient with von Recklinghausen's neurofibromatosis. Report of a case. AB - A 42-year-old woman complicated with neurofibromatosis underwent both balloon percutaneous transluminal angioplasty (PTA) of the iliac artery and femoropopliteal (FP) bypass grafting for critical lower limb ischemia. Seven months after the initial intervention, a recurrence of stenosis in the iliac artery and at the anastomoses of the FP bypass necessitated both PTA and a repeat thrombectomy and finally resulted in the amputation of her left thigh. This is a rarely documented case of chronic arterial occlusion associated with neurofibromatosis, in which the prognosis of arterial reconstruction for such patients is suggested to be poor. PMID- 11052297 TI - Upper limb ischemia induced by chronic licorice ingestion. AB - A young woman had an acute embolic episode that affected her right upper limb. An axillo-brachial thromboembolectomy was performed after which distal pulses were recovered. The patient had a long lasting history, longer than ten years, of continuous licorice ingestion. Blood samples showed severe hypokalemia that caused EKG changes. Transesophageal echocardiogram discovered mild mitral valve prolapse. The combination of surgery and potassium supplements was successful in treating this rare and potentially life-threatening disease. PMID- 11052298 TI - Pleurectomy in primary pneumothorax: is extensive pleurectomy necessary? AB - BACKGROUND: The aim of the study was to evaluate the results of parietal pleurectomy in patients with primary spontaneous pneumothorax comparing extensive pleurectomy performed by thoracotomy versus more limited pleurectomy performed by VATS. METHODS: Records of the patients operated on for primary pneumothorax at Royal Brompton Hospital from January 1994 to April 1997 were retrospectively reviewed. A follow-up questionnaire was sent to patients asking about further pneumothorax and the presence of long-term chest problems on the operated side. A statistical uni- and multivariate analysis was performed searching predictors for postoperative complications, recurrence and chronic chest problems. RESULTS: Thirty-six patients underwent extensive pleurectomy through a limited postero lateral thoracotomy (40%, group A), 54 patients had a limited pleurectomy (60%, group B), 50 by VATS and 4 by axillary thoracotomy. Overall, 11 patients had postoperative complications (12.2%). In group A, 4 patients (11.1%) had complications (2 reoperation, 2 air leak >7 days). In group B, 7 patients (12.9%) had complications (1 reoperation, air leak >7 days, 1 wound infection). Two patients experienced recurrent ipsilateral pneumothorax after surgery, both belonging to group B (overall recurrence rate 2.5%, group B 4.1%). Thirteen patients in both groups (respectively 41.9% in group A and 27% in group B) admitted chest problems on the operated side. From statistical analysis, "indication" resulted a predictor of complications (p=0.03) and "thoracotomy" a predictor of long-term chest problems (p=0.03). CONCLUSIONS: Many theoretical advantages of limited VATS pleurectomy have still to be confirmed and it is reasonable to use it in uncomplicated primary pneumothorax. The superb exposure obtained with thoracotomy and the superiority of extensive pleurectomy in terms of recurrence indicate this approach in case of complicated pneumothorax or when long-term security is of paramount importance. PMID- 11052299 TI - Morbidity and survival after bronchoplastic surgery for non-small-cell lung cancer. AB - BACKGROUND: Bronchoplastic procedures are an accepted surgical approach in patients with resectable non-small-cell lung cancer (NSCLC) to avoid pneumonectomy. Post-operative complications associated with the bronchial anastomosis and local recurrence of the tumor have to be considered. Experimental design and setting: Retrospective analysis of the clinical courses and follow-up of 1,610 consecutive patients who received surgical resection for NSCLC at the Department of Surgery, Klinikum Grosshadern, University of Munich, Germany. Among them there were 134 (8.3%) bronchoplastic resections. METHODS: Morbidity, mortality, and survival rate were investigated in these patients to verify the safety of this technique. RESULTS: From all 134 bronchoplastic resections, 105 lobectomies, 22 bilobectomies, and 7 pneumonectumies were performed. Atelectasis was observed in 6.0% (versus conventional procedures: 3.7%; p: n.s.), whereas anastomotic dehiscence occurred in 3.0%. In-hospital mortality amounted to 3.7% (versus 5.3%; p: n.s.). The stage dependent 5-year survival in R0-resected patients was comparable in both groups. CONCLUSIONS: Our results demonstrate that bronchoplastic procedures represent a safe therapeutic option in the operative treatment of non-small-cell lung cancer that should be considered in all patients with central tumor growth. PMID- 11052300 TI - Spontaneous intercostal pulmonary hernia. AB - A case of spontaneous intercostal pulmonary hernia as a result of vigorous coughing is reported in a 67-year-old man. The great majority of acquired pulmonary hernias are post-traumatic; rare cases are spontaneous, resulting from prolonged and/or repeated increased intrathoracic pressure. This hernia was successfully repaired with a polyglactin absorbable mesh and approximation of the ribs with heavy stitches. When required, surgical repair is the treatment of choice. PMID- 11052301 TI - Anterior diaphragmatic hernia misinterpreted by X-ray, echocardiography, computed tomography scanning and magnetic resonance imaging. AB - We report the case of a 23-year-old man who was admitted to our Division with the diagnosis of pericardial lipoma. Chest X-ray, echocardiography and magnetic resonance imaging failed to reveal an anterior diaphragmatic hernia containing a small part of the stomach with a big prehernial lipoma that were found at surgery. We believe that in all cases of suspected pericardial lipoma a diaphragmatic hernia should be expected. PMID- 11052302 TI - About sinus of Valsalva aneurysm. PMID- 11052303 TI - Inflammatory aneurysms of the abdominal aorta. A prospective clinical study. PMID- 11052304 TI - Transesophageal echocardiography in the treatment of right atrial hydatid cyst. Its impact on diagnosis and surgical management. PMID- 11052305 TI - Combined internal carotid and primitive hypoglossal artery endarterectomy. PMID- 11052306 TI - Variations in digestive physiology of rats after short duration flights aboard the US space shuttle. AB - The purpose of this work was to assess the influence of microgravity on several endogenous and microbial parameters of digestive physiology. On the occasion of two Spacelab Life Sciences missions, SLS-1 (a 9-day space flight) and SLS-2 (a 14 day space flight), Sprague-Dawley rats flown aboard the US space shuttle were compared to age-matched ground-based controls. In both flights, exposure to microgravity modified cecal fermentation: concentration and profile of short chain fatty acids were altered, whereas urea and ammonia remained unchanged. Only in SLS-1 was there an induction of intestinal glutathione-S-transferase. Additional analyses in SLS-2 showed a decrease of hepatic CYP450 and of colonic goblet cells containing neutral mucin. After a postflight recovery period equal to the mission length, only modifications of the hepatic and intestinal xenobiotic metabolizing enzymes still persisted. These findings should help to predict the alterations of digestive physiology and detoxification potential likely to occur in astronauts. Their possible influence on health is discussed. PMID- 11052307 TI - PYY regulates pancreatic exocrine secretion through multiple receptors in the awake rat. AB - Peptide YY (PYY) is one of several regulatory peptides reported to modulate pancreatic secretion. PYY circulates in two forms, PYY1-36 and PYY3-36, and binds to multiple receptor subtypes. We sought to determine if PYY1-36 or PYY3-36 regulates neurally mediated pancreatic secretion through the Y1, Y2, and/or Y5 receptor subtypes. Experiments were conducted in awake, surgically recovered rats. In order to determine the effects of the PYYs on basal pancreatic secretion, either PYY1-36, [Pro34] PYY1-36 (a Y1/Y5 agonist), or PYY3-36 (a Y2/Y5 agonist) were infused for 40 min at doses of 0, 12.5, 25, or 50 pmol/kg/hr while measuring pancreatic juice volume and protein. PYY1-36 increased pancreatic protein secretion at 25 and 50 pmol/kg/hr (P < 0.05) in a dose-dependent manner (P < 0.001, R2 = 0.990). The Y2/Y5 receptor agonist PYY3-36 significantly inhibited pancreatic juice volume and protein at 12.5 and 25 pmol/kg/hr, but stimulated protein secretion at higher doses (P < 0.001, R2 = 0.995). The Y1/Y5 agonist, [Pro34] PYY1-36, had no significant effect on basal pancreatic exocrine secretion. Therefore, PYY1-36, PYY3-36 and [Pro34] PYY1-36 produced different, dose-dependent changes on basal pancreatic exocrine secretion. Inhibition of pancreatic secretion by circulating PYY1-36 and PYY3-36 are primarily mediated by the Y2 receptor. Since [Pro34] PYY1-36 did not change pancreatic secretion, it can be concluded that circulating PYY1-36 or PYY3-36 does not modulate pancreatic secretion through the Y1 or Y5 receptors. Since the stimulatory effects of PYY1 36 on pancreatic secretion could not be explained by the actions of PYY3-36 or [Pro34] PYY1-36 on Y1 or Y2 receptors, and since PYY1-36 fails to bind to Y3 or Y4 receptors, we also conclude that PYY1-36 may stimulate pancreatic secretion in a dose-dependent mechanism through a PYY receptor subtype different from Y1, Y2, Y3, Y4 or Y5. PMID- 11052308 TI - Operative stress response is reduced after laparoscopic compared to open cholecystectomy: the relationship with postoperative pain and ileus. AB - Our objective was to determine the least invasive surgical procedure; to do this we compared postoperative pain, duration of ileus, and level of neurohormonal stress response after laparoscopic cholecystectomy (LC) and open cholecystectomy (OC). Postoperative recovery of patients was faster after LC than OC but comparison of the neurohormonal stress response after laparoscopic and open surgical procedures revealed conflicting results. Forty-one consecutive patients with noncomplicated gallstones were randomized for LC (N = 25) and OC (N = 16). The stress level was evaluated in patients before surgery by the Hamilton anxiety scale. Postoperative pain was assessed by a visual analogic scale (VAS) pain score and by the amount of analgesic drugs (propacetamol) administered, while the duration of ileus was determined by the delay between surgery and the time to first passage of flatus as well by the colonic transit time (CTT) measured by radiopaque markers. Plasma concentrations of anti-diuretic hormone (ADH), adrenocorticotropic hormone (ACTH), beta-endorphin (BE), neurotensin (NT), and aldosterone (Ald) were measured before and during surgery as well as 2 and 5 hr after the surgery (D0) and on the day following surgery (D1). Urinary cortisol (uCOR) and urinary catecholamine metabolites were assessed before surgery, during D0, and on D1. Patient characteristics, the duration of surgery, and the doses of anesthetic drugs were not different in LC and OC. In LC patients the VAS pain score and the doses of postoperative antalgics were lower (P < 0.05), the time to first passage of flatus was shorter (P < 0.001), and the CTT tended to be shorter (54 +/- 12 hr vs 81 +/- 17) compared to OC patients. Patients who required the highest doses of postoperative antalgics had the longest delay to first passage of flatus (P < 0.01). During surgery, all neurohormonal parameters increased compared to the preoperative period (P < 0.05), and only plasma NT concentrations were lower during LC than OC (P < 0.05). During the postoperative period, ACTH, BE, Aid, catecholamines, and uCOR concentrations were lower in LC than in OC (P < 0.05). Concentrations of hormonal parameters were higher when the duration of surgery increased (P < 0.05). A greater need for propacetamol to relieve pain was associated with a greater increase in BE, ACTH, and urinary catecholamine levels (P < 0.05-P < 0.005). When the time to first passage of flatus was delayed, levels of BE, ACTH, and catecholamines and NT concentrations were increased (P < 0.05-P < 0.005). In conclusion, LC is less invasive because this surgical procedure induces a shorter neurohormonal stress response than OC, even if the peroperative response is not different. Postoperative pain levels and the duration of ileus are associated with BE, ACTH, and catecholamine levels and NT concentrations, suggesting the importance of hormones in postoperative functional recovery. PMID- 11052309 TI - Sphincter of Oddi motility in patients with hepatolithiasis and common bile duct stones. AB - The purpose of this study was to explore a difference in sphincter of Oddi (SO) motor activity among patients with intrahepatic (I, N = 5), intra- and extrahepatic (IE, N = 15), and common bile duct (CBD, N = 6) stones. Interdigestive motility of the SO and duodenum was studied by pneumohydraulic infusion manometry via the percutaneous route. SO phasic contractions showed a cyclic change in concert with the duodenal migrating motor complex (MMC) in all these patients. There was no significant difference in the cycle length, frequency, or amplitude of the SO phasic waves among the three groups throughout the whole cycle. The SO basal pressure during duodenal phases I and II of the duodenal MMC was significantly lower in patients with the IE type of hepatolithiasis than in those with the I type (P = 0.04), but there was no significant difference during phase III between the two groups. The SO basal pressure during phases I and II of the CBD group was also significantly lower than that of the I group (P = 0.02). The significance became even more prominent (P = 0.001) when a subgroup of patients with a dilated CBD (diameter > 1 cm) was examined. Lower basal pressure in the IE group or CBD group than in the I group suggested that stones in the common duct might injure or irritate the SO and cause SO dysfunction. In the subgroup with dilated CBD, which may have resulted from repeated and severe SO injury, the statistics became more prominent. PMID- 11052310 TI - Effects of very long chain versus long chain triglycerides on gastrointestinal motility and hormone release in humans. AB - Fish oil (a very long chain triglycerides, VLCT) has received much attention because of its favorable metabolic properties; however, its effect on gastrointestinal function has not been studied. We investigated the effects of intraduodenally administered VLCT on gut-hormone release [cholecystokinin (CCK), neurotensin, peptide YY (PYY)], gallbladder emptying, antroduodenal motility, and small bowel transit time (SBTT) in comparison to intraduodenal administration of saline and long chain triglycerides (LCT, corn oil) in nine healthy volunteers. Gallbladder contraction duration was significantly shorter after VLCT than after LCT (138 +/- 16 min vs 233 +/- 38 min, P < 0.05). Both fats induced a fed motility pattern, while SBTT was not significantly altered. CCK secretion was significantly reduced after VLCT compared to LCT (36 +/- 12 pM x 120 min vs 78 +/ 15 pM x 120 min, P < 0.05), whereas PYY and neurotensin release were not significantly different. In conclusion, effects of triglycerides on CCK and gallbladder motility appear to be chain-length dependent, in contrast to the effects on distal gut-hormone release and intestinal motility and transit, which appear to be chain-length independent. PMID- 11052311 TI - Assessment of gastric electrical activity and autonomic function among diabetic and nondiabetic patients with symptoms of gastroesophageal reflux. AB - Gastroesophageal reflux disease (GERD) may present differently in patients with diabetes mellitus (DM) than in nondiabetics (NDM). We compared three tests in two patient groups with GERD symptoms: a DM group (n = 10) and a NDM group (n = 13). The tests were 24-hr esophageal pH, autonomic function testing (AFT), and electrogastrography (EGG). Analysis of the 23 patients revealed the DM group had normal 24-hr pH values (9 of 10 patients, mean pH 3.1 +/- 1.7), while NDM displayed abnormal pH values (9 of 13 patients, mean pH 21.2 +/- 5.9). AFT results were abnormal in DM (demonstrating cholinergic/ adrenergic dysfunction), but normal in NDM. EGG values were abnormal in both groups (mean 3.31 +/- 0.1 in each). We conclude that in GERD-symptomatic patients, those with DM frequently have normal 24-hr pH, but abnormal autonomic functioning, in contrast to NDM, who have abnormal 24-hr pH but normal autonomic function. Both groups had identically abnormal mean EGG values. PMID- 11052312 TI - Impairment of gastric and jejunal myoelectrical activity during rectal distension in dogs. AB - It is known that distension of the rectum induces gastric hypomotility and delays gastric emptying. Its effect on gastrointestinal myoelectrical activity has not been well studied, however. The aim of this study was to investigate the effects of rectal distension on gastrointestinal myoelectrical activity in dogs. Six hound dogs implanted with electrodes on the serosa of the stomach and proximal jejunum were studied. The protocol consisted of a 30-min baseline recording and another 30-min recording during rectal distension. Gastric myoelectrical activity was severely impaired by rectal distension. The dominant power was significantly decreased from -2.79 +/- 0.52 dB at baseline to -4.84 +/- 1.26 dB during distension (P < 0.05). The percentage of normal 4-6 cycles per minute gastric slow waves was reduced from 95.08 +/- 1.11% to 83.63 +/- 4.00% (P < 0.02), and the percentage of tachygastria was increased during distension (0.33 +/- 0.19% vs 6.03 +/- 1.27%, P < 0.02). The instability coefficient of the dominant frequency was significantly increased (0.134 +/- 0.012 vs 0.326 +/- 0.074, P < 0.05). The percentage of slow wave coupling was reduced from 93.99 +/- 0.76% to 73.43 +/- 2.07% (P < 0.00003). In the small bowel, only the instability coefficient of dominant frequency showed a significant increase during distension. Other parameters were not affected by rectal distension. We conclude that rectal distension severely impairs gastric myoelectrical activity. The induced gastric dysrhythmia and reduced slow wave amplitude and coupling may be the underlying pathophysiology of gastric hypomotility and delayed gastric emptying observed during rectal distension. PMID- 11052313 TI - A tailored perioperative blood transfusion might avoid undue recurrences in gastric carcinoma patients. AB - In a large retrospective study, the prognostic significance of blood transfusion (BT) and type of blood product transfused were searched in 640 curatively resected gastric carcinoma patients. Out of 640 patients, 222 (34.7%) received BT and the incidence of BT was reduced from 45% in the early period to 21% in the most recent period. The five-year disease-free survival was significantly (P < 0.0001) worse in the transfused group and BT became an independent prognosticator (P = 0.0061) in the multivariate analysis. Patients transfused with only packed red blood cells (PRBC) had significantly (P = 0.0258) better survival than those transfused with other types of blood products. In conclusion, BT was found to be an independent prognosticator in gastric carcinoma patients after curative resection. A tailored transfusion with PRBC, when indicated, might be the choice for better survival than whole blood or other blood products. PMID- 11052314 TI - Pheasant-induced dysphagia. PMID- 11052315 TI - Transmission of Helicobacter pylori from challenged to nonchallenged nude mice kept in a single cage. AB - To determine the transmission route of Helicobacter pylori, one nude mouse was challenged by H. pylori, and then raised with nonchallenged nude mice in a single cage in a sterilized environment with and without exposure to their feces. After coraising for two and four weeks, all mice were killed to determine H. pylori in the stomach, saliva, and feces and to assess gastritis grade. Natural transmission of H. pylori occurred in 50% (2/4) and 70% (7/10) of mice after two weeks and four weeks of coraising when they were exposed to their feces. H. pylori was detected not only in the stomach but also in saliva and feces by PCR of all challenged and transmitted mice. However, no transmission occurred in mice not exposed to feces of a challenged mouse, while sharing food and water in a single cage. These findings suggest that the fecal-oral transmission route is important, at least in the animal model. Serum levels of anti-H. pylori urease IgG of the H. pylori-transmitted mice increased after coraising, and gastritis was observed in the stomach of both challenged and transmitted mice. We conclude that H. pylori bacteria are transmitted through the fecal-oral route from challenged to nonchallenged nude mice, resulting in gastritis. PMID- 11052316 TI - Long-term effects of Helicobacter pylori eradication on intestinal metaplasia in patients with duodenal and benign gastric ulcers. AB - This study was conducted to investigate whether or not the eradication of H. pylori could lead to the regression of intestinal metaplasia (IM) in patients with either duodenal ulcer (DU) or benign gastric ulcer (BGU). The initial antral IM grade was 0.21 in the 72 patients of the H. pylori-eradicated DU group, this decreased to 0.17, 0.14, 0.13, and 0.09 after periods of four weeks, one year, two years, and four years, respectively, but without statistical significance. In the corpus of the DU group, where IM grade was low (0.02), there was no detectable change in IM. The initial antral IM grade of 0.69 in the 41 patients of the H. pylori-eradicated BGU group decreased substantially to 0.61, 0.44, and 0.39 after periods of four weeks and one and two years, respectively, but again without statistical significance. The initial corporal IM grade of the BGU group of 0.27 decreased to 0.20, 0.15, and 0.06 after periods of four weeks and one and two years, again without statistical significance. In contrast, the IM grades of the noneradicated DU group (N = 20) and the BGU group (N = 16) showed nearly no change in the antrum and corpus. Gastritis grades of antrum and corpus in the H. pylori-eradicated DU or BGU group significantly decreased with respect to time (P = 0.0001), but there were no significant changes in the corresponding noneradicated groups. Although there was no statistical significance, IM decreased in the antrum and corpus of the stomach with BGU and in antrum of those with DU over a two to four-year period after H. pylori eradication, suggesting the possible reversibility of IM. PMID- 11052317 TI - Long-term follow-up study of gastric emptying and Helicobacter pylori eradication among patients with functional dyspepsia. AB - Studies on the influence of Helicobacter pylori gastritis on gastric motility have produced inconclusive results. We investigated the effect of Helicobacter pylori eradication therapy on gastric emptying in patients with functional dyspepsia in a placebo-controlled double-blind study with one year follow-up. A standardized scintigraphic double-tracer examination was used. Of the 40 subjects, 29 were H. pylori-positive patients with functional dyspepsia and 11 were asymptomatic control subjects. Gastric emptying parameters were: postlag 50% retention time for solids (T50), gastric emptying half-time for liquids (T1/2), solid lag duration, and intragastric distribution of solids. At baseline, the scintigraphic examination was performed for all study subjects to detect any major alterations between dyspeptic patients and asymptomatic control subjects. Thereafter every patient was randomized to receive either H. pylori eradication therapy or placebo; in addition they also received omeprazole 20 mg once a day for three months to stabilize the acid suppression therapy. After one year scintigraphy was repeated for the patients. The solid lagtime was prolonged among dyspeptic patients compared with asymptomatic controls (P = 0.02). After one year there was no significant difference between H. pylori-eradicated and placebo treated patients in any gastric emptying parameter. However, good reproducibility of the scintigraphic examination showing the gastric emptying rate of solids (r = 0.43, 95% CI: 0.07-0.69; P = 0.02) and liquids (r = 0.44, 95% CI: 0.09-0.69; P = 0.02) continued even after one year of follow-up. In conclusion, eradication of H. pylori has no impact on gastric emptying in patients with functional dyspepsia, but the long-term trend in individual gastric emptying rate is stable. PMID- 11052318 TI - Helicobacter pylori reduces intracellular glutathione in gastric epithelial cells. AB - Helicobacter pylori infection has been associated with stimulation of gastric mucosal reactive oxygen species (ROS) production, and it was postulated that ROS production is due to neutrophil infiltration and activation. The aim of this study was to investigate the direct effect of H. pylori on ROS formation in gastric epithelial cells in vitro. The human gastric cancer cell line HM02 was incubated with H. pylori for 24 hr, and the effects on cell number and the intracellular radical scavenger reduced glutathione (GSH) were assessed. H. pylori caused a concentration-dependent reduction of cellular GSH concentrations over a broad bacteria-to-cell ratio (1.4-42) in the absence of cell necrosis. The radical scavengers MnTBAP (a cell permeable superoxide dismutase) and ebselen provided protection against H. pylori-induced decrease in cellular GSH concentrations. We conclude that H. pylori directly decreases cellular GSH concentrations in gastric epithelial cells. We suggest that this effect is caused by the release of ROS by H. pylori. PMID- 11052319 TI - Cytoskeletal rearrangements induced by Helicobacter pylori strains in epithelial cell culture: possible role of the cytotoxin. AB - The relationship between Helicobacter pylori adherence, cytotoxin production, and modification of the cytoskeletal structure was investigated by studying the effects of 12 H. pylori strains cocultured with Hep-2 epithelial cells. Bacterial strains were isolated from patients with peptic ulcer disease or nonulcer dyspepsia. Presence of the cag pathogenicity island and vacA subtypes of the strains were determined as was the production of vacuolating cytotoxin. We found that cytoskeletal rearrangements, as observed by confocal microscopy after double staining of the bacteria and the cell actin with Texas red and fluorescein conjugated phalloidin, respectively, occurred essentially when the strains were cytotoxin producers and that the supernatants alone could also lead to these modifications. PMID- 11052320 TI - Cytomegalovirus infection of gastrointestinal tract with multiple ulcers and strictures, causing obstruction in a patient with common variable immunodeficiency syndrome. PMID- 11052321 TI - Suppressive effect of antiulcer agents on granulocytes--a role for granulocytes in gastric ulcer formation. AB - Many clinicians have believed that H2-blockers and proton pump inhibitors ameliorate gastric ulcers via their antacid function. We examined the effects of these antacids on granulocytes. Gastric ulcer patients were administered an H2 blocker or proton pump inhibitor for a week and the number of granulocytes and the superoxide production were examined. To determine the trafficking of granulocytes, mice were exposed to restraint stress for 24 hr. The H2-blocker decreased the number of granulocytes, while the proton pump inhibitor suppressed their superoxide production in humans and mice. The major function of H2-blockers and proton pump inhibitors in curing gastric ulcers seems to be their suppressive effects on granulocytes. In this case, stress accelerates the trafficking of granulocytes from the bone marrow to the gastric mucosa. If we demonstrate a role for granulocytes in gastric ulcer formation, an gap in the acid-pepsin theory and the Helicobacter pylori theory is filled in. PMID- 11052322 TI - Effect of bisphosphonates on surface hydrophobicity and phosphatidylcholine concentration of rodent gastric mucosa. AB - Bisphosphonates are a family of chemically related zwitterionic molecules that are used clinically to retard bone resorption in individuals with osteoporosis and associated skeletal diseases. Inflammation and ulceration of the upper gastrointestinal tract by a mechanism that relates to a topical irritant action is associated with the consumption of some bisphosphonates. In the present study, we investigated the effects of three bisphosphonate molecules, pamidronate, alendronate, and risedronate on the surface hydrophobicity and phosphatidylcholine (PC) concentration of the antral mucosa. We also examined how these surface changes related to mucosal injury in an established rat model, in which the test compounds were administered in combination with indomethacin. We initially determined that a combination of pamidronate (300 mg/kg) and indomethacin (40 mg/kg) induced a significant reduction in mucosal surface hydrophobicity and macroscopic lesion formation by 15 min and mucosal PC concentration by 30 min, with the magnitude of these changes increasing over the 4-hr study period. An equivalent dose of alendronate or risedronate in combination with indomethacin produced modest or no macroscopic injury, respectively, to the antral mucosa over the 4-hr study, although the bisphosphonates clearly induced surface injury and some glandular necrosis when examined at the light microscopic level. These bisphosphonates also induced modest decreases in antral surface hydrophobicity and mucosal PC concentration that appeared to be related to their injurious potential. In conclusion, the variable toxicity of bisphosphonates to the antral mucosa appears to be associated with their ability to compromise the surface hydrophobic phospholipid barrier of the tissue, with pamidronate > > > alendronate > risedronate. This bisphosphonate effect on the surface barrier may trigger the development of mucosal injury and possible ulceration. PMID- 11052324 TI - Use of 6-mercaptopurine in patients with inflammatory bowel disease previously intolerant of azathioprine. AB - Both azathioprine and its active metabolite, 6-mercaptopurine, are of benefit in the treatment of inflammatory bowel disease, either in resistant cases, or for their steroid-sparing effect. Azathioprine treatment is limited in some patients by hypersensitivity reactions or other side effects. We report our experience in 11 patients previously unable to tolerate azathioprine for a variety of reasons, who were switched to 6-mercaptopurine. Of seven patients with ulcerative colitis and four patients with Crohn's disease who were treated with 6-mercaptopurine following failed azathioprine therapy, six were able to successfully tolerate the substitute medication, with good response. Where patients have previously been intolerant of azathioprine yet ongoing indications for immunosuppressive therapy remain, a trial of 6-mercatopurine may be warranted. Given the similar efficacies of the two drugs in inflammatory bowel disease, these findings also favor the use of 6-mercaptopurine rather than the parent compound as initial therapy. PMID- 11052323 TI - Nitric oxide and superoxide anion in low-grade esophagitis induced by acid and pepsin in rabbits. AB - It has been suggested that free radicals are involved in esophagitis. To study the role and potential interaction of superoxide anion and nitric oxide (NO) in low-grade esophagitis, we perfused acidified pepsin (30 min every 12 hr) for seven days in rabbits treated with different agents to modulate the generation of these radicals. Measurements included macroscopic and microscopic damage, superoxide anion generation, mucosal nitric oxide synthase activity, and peroxynitrite formation. Low-grade esophagitis was associated with increased nitric oxide synthase mucosal activity and mucosal damage was dose-dependently increased by treatment with the NO synthase inhibitor NG-nitro-L-arginine. Superoxide anion was scarcely generated in the mucosa, but this was not accompanied by any change in the activity of mucosal superoxide dismutase. Treatment with superoxide dismutase did not improve mucosal damage. Generation of peroxynitrites was not detected. In conclusion, nitric oxide is involved in the mucosal defense of the esophagus against acid- and pepsin-induced damage. Superoxide anion generation seems irrelevant in the induction of low-grade esophagitis and not sufficient to interact with nitric oxide to generate measurable mucosal peroxynitrite radicals. PMID- 11052325 TI - Total antioxidant capacity of colon in patients with chronic ulcerative colitis. AB - It has been proposed that oxidative stress is involved in the pathophysiology of ulcerative colitis. We have reported the depletion of the nonenzymatic antioxidant, glutathione, in colon from active and inactive ulcerative colitis. The colon contains several biochemically linked antioxidant systems. We hypothesized that diminished total antioxidant capacity in active ulcerative colitis would be associated with increased colonic lipid peroxidation. This study was designed to determine total antioxidant capacity and lipid hydroperoxide levels using colon obtained at surgery from controls (N = 16; 4 females, 12 males; mean age 70 years), and active and inactive ulcerative colitis (N = 15; 3 females, 12 males; mean age 39). Total antioxidant capacity of control colon was higher in muscularis externa compared to the mucosal-submucosal layer (P < 0.05). There were no differences in colonic total antioxidant capacity or lipid hydroperoxide levels comparing control colon to inactive and active ulcerative colitis. The results did not support depletion of tissue total antioxidant capacity by free radicals. Depletion of glutathione in ulcerative colitis may be a specific disorder rather than a secondary defect attributable to global oxidative stress. Nonspecific antioxidant supplements appear unlikely to be beneficial in the treatment of ulcerative colitis. PMID- 11052326 TI - Experimental ulcerative colitis impairs antioxidant defense system in rat intestine. AB - Increasing attention has been given recently to the role of free radicals in the pathogenesis of ulcerative colitis, since the inflamed intestine is exposed to oxidative stress generated by infiltrating macrophages and neutrophils within the lamina propia. The overall goal of this study was to evaluate whether experimental ulcerative colitis induces significant changes in the antioxidant defense system in an experimental model induced by the intrarectal administration of 2,4,6-trinitrobenzenesulfonic acid. Twenty rats were treated with 80 mg/kg body weight of trinitrobenzenesulfonic acid and 20 with the same volume of 0.9% NaCl. Rats were killed at one and two weeks after treatment to evaluate colon damage by light and electron transmission microscopy. The degree of tissue injury and inflammation was determined by measuring alkaline phosphatase, gamma glutamyltranspeptidase, and myeloperoxidase activities and prostaglandin E2 and leukotriene B4. Glutathione levels and the activity of the enzymes of the antioxidant defense system were determined. Enzymatic markers of colon injury showed higher activities in rats with ulcerative colitis. Concentrations of prostaglandin E2 and leukotriene B4 were higher in the groups treated for one week with trinitrobenzenesulfonic acid and markers decreased after two weeks of treatment. All antioxidant enzyme activities were higher at one and two weeks after treatment; however, a significant decrease in total glutathione content was also observed. In conclusion, ulcerative colitis induced by trinitrobenzenesulfonic acid damages the intestinal mucosa and is accompanied by a shift in the antioxidant enzyme activities, and low levels of glutathione. This deficiency in glutathione could be a target for new therapies to treat ulcerative colitis. PMID- 11052327 TI - Optimization of technetium-99m-HMPAO leukocyte scintigraphy in evaluation of active inflammatory bowel disease. AB - Although [99mTc]HMPAO-labeled leukocyte scintigraphy is widely used in the assessment of IBD, the time point chosen for imaging is still controversial. The aim of the present study was to determine the optimal scanning sequence to assess IBD extension and activity. Sixty-two consecutive patients with active and 18 with inactive IBD were prospectively studied. Clinical evaluation, colonoscopy, radiology, and scintigraphy were performed within three days, without changes in the patient's treatment. Compared to early scan (45 min), late scan (3 hr) had a higher sensitivity (85% vs 100%) and accuracy (85% vs 95%) in identifying patients with active IBD and in defining IBD extension. Combinations of values from both scans did not improve accuracy of scintigraphy, which is lower in Crohn's disease than in ulcerative colitis and also in patients receiving steroid treatment. In conclusion, a single late scintigraphy scan provides the best means to identify patients with active IBD and to assess disease extension. PMID- 11052328 TI - Colonic perforation in unsuspected amebic colitis. AB - Unsuspected amebic colitis presenting as inflammatory bowel disease, as in our patient, has been previously reported (4, 7, 8). Misdiagnosis, delay in antibiotic treatment, and institution of immunosuppression were the result of failure to identify the parasite in stool specimens and have resulted in suffering, morbidity, mortality, and surgery. In all previously reported cases, routine stool studies failed to identify E. histolytica (4, 7, 8). The correct diagnosis was only established after reviewing the surgical specimen or biopsies obtained endoscopically. Because the erroneous diagnosis of inflammatory bowel disease can lead to disastrous complications, it is imperative to exclude amebic colitis prior to undertaking steroid therapy, especially in patients with a prior history of travel to or residence in areas with endemic E. histolytica (17). We recommend obtaining at least three stool specimens for microscopic examination, as well as testing for serum amebic antibody. Patients should submit fresh stool specimens directly to the laboratory to allow for prompt diagnostic evaluation. Such an approach might lead to the improved diagnosis of amebiasis. PMID- 11052329 TI - Chronic ingestion of a potential food contaminant induces gastrointestinal inflammation in rats: role of nitric oxide and mast cells. AB - Chronic ingestion of xenobiotics could be pathogenic in the gastrointestinal tract. Recently, we showed that acute low administration of a food contaminant (diquat) induced intestinal secretion involving mast cells and nitric oxide. This work aimed to determine in rats: (1) the influence of a low level (0.1 mg/kg/day per os) chronic ingestion of diquat on gastrointestinal immune cells, and (2) the participation of nitric oxide synthases (NOS) in these effects. Diquat increased both gastric and jejunal myeloperoxidase activities, tissue histamine in vitro release after stimulation by 48/80, and mast cell numbers. Diquat did not alter gastric NOS but increased intestinal inducible NOS (iNOS) activity. L-NAME prevented diquat-induced gastric and intestinal mastocytosis and gastric but not intestinal inflammation. L-NAME reduced gastric constitutive NOS (cNOS) activity and reestablished control iNOS activity. Chronic low level ingestion of diquat induces a low-grade gastric and intestinal inflammation with mastocytosis and enhancement of intestinal iNOS activity. PMID- 11052330 TI - Localized amyloid tumor in small bowel. AB - We report a case of localized jejunal amyloidosis occurring in a 74-year-old man who experienced an episode of digestive bleeding while he was receiving oral anticoagulation. It illustrates a rare entity, characterized by an endoscopic aspect of polypoid, pseudotumoral formations. Histologically, submucosal connective tissues, muscularis mucosae, and blood vessel walls are massively infiltrated by amyloid, giving a typical red/green birefringence under polarized light. PMID- 11052331 TI - Hepatitis C virus infection and lymphoproliferative disorders. PMID- 11052332 TI - Clinical significance of patent paraumbilical vein in patients with liver cirrhosis. AB - A patent paraumbilical vein (PUV) is a frequent finding in patients with cirrhosis when studied by duplex Doppler ultrasound. There is controversy regarding the clinical significance of this finding. We studied 50 patients with cirrhosis and portal hypertension as evidenced by the demonstration of esophageal varices on endoscopy. All 50 patients were evaluated for a significant PUV (diameter of > or =3 mm) using duplex Doppler sonography. The patients were divided into two groups based on the size of esophageal varices (group A with small varices, N = 30; group B with large varices, N = 20). A significant PUV was seen in 21 (42%) patients. The patients with portosystemic encephalopathy had a significantly greater prevalence of PUV (70%) than those without (32%, P < 0.02). Of the group with large esophageal varices, 6 (30%) had a significant PUV, while in the group with small varices, 15 (50%) had a significant PUV (P > 0.05). In no patient with large varices did the PUV diameter exceed 6 mm, while in as many as six patients with small varices, the PUV diameter exceeded 6 mm. A significantly enlarged PUV seen on duplex Doppler ultrasound in cirrhotics may have important hemodynamic consequences. Besides predisposing the patient to portosystemic encephalopathy, it may also offer some protection against formation of large varices. In particular, a very large patent PUV (> or =6 mm) might identify a subgroup of patients with small varices and thus a lesser likelihood of variceal bleeding. PMID- 11052333 TI - Increased plasma levels of corticosterone and prolactin and decreased T3 and T4 levels in short-term prehepatic portal hypertension in rats. AB - Corticosterone, T3, T4, and prolactin serum concentrations at 24 hr (N = 10), 15 days (N = 10), and 45 days (N = 10) of postoperative (postop) evolution were assayed to study the neuroendocrine response to portal hypertension. A triple stenosing ligature of the portal vein was used as the surgical technique of portal hypertension. This technique does not produce mortality and causes a decrease in the serum concentrations of T3 (0.043 +/- 0.009 vs 0.55 +/- 0.08 ng/ml) and T4 (3.93 +/- 0.55 vs 4.65 +/- 0.67 microg/ml) and an increase in those of prolactin (28.61 +/- 20.20 vs 12.84 +/- 3.96 ng/ml) and corticosterone (397.50 +/- 64.17 vs 311.53 +/- 57.41 ng/ml) at 45 days postop. The T3, T4, prolactin, and corticosterone alterations are associated with a persistent increase of TNF alpha and NO, whose serum concentrations at 45 days postop are, respectively, 1838.33 +/- 247.07 vs 48.89 +/- 8.75 pg/ml and 0.43 +/- 0.13 vs 0.19 +/- 0.01 mmol/ml. TNF-alpha and NO could mediate these hormonal alterations in the evolution of short-term portal hypertension in the rat; thus they are involved in the systemic neuroendocrine response that is induced by this injury. PMID- 11052334 TI - Acute hepatitis induced by bupropion. AB - As an antidepressant, bupropion is considered to be a safe agent that usually causes infrequent and mild increase of serum liver enzymes. Asymptomatic elevation of serum transaminases was previously reported only in a single case. We describe a patient who developed typical acute hepatitis after receiving six weeks of bupropion for depression. His presentation was characterized with acute onset of symptoms associated with significantly elevated ALT, AST, and LDH and acute hepatic inflammation. The clinical course of our patient, including incubation period, pattern of liver enzyme elevation, and time of recovery, was similar to, but much more severe than, the case reported by Oslin and Duffy. Discontinuation of bupropion was followed by a rapid resolution of clinical symptoms and liver enzymes. The incidence of bupropion-induced hepatitis remains to be defined even though it appears to be relatively low. Since the clinical application of bupropion is broader, we must be aware of the clinical entity of bupropion-induced hepatitis. PMID- 11052335 TI - Acute hepatitis after lamotrigine administration. PMID- 11052336 TI - Autoimmune hepatitis associated with autoimmune hemolytic anemia and anticardiolipin antibody syndrome. PMID- 11052337 TI - Tolcapone-related fulminant hepatitis: electron microscopy shows mitochondrial alterations. PMID- 11052338 TI - Deposition of eosinophil granule proteins in liver associated with allergic bronchopulmonary candidiasis. PMID- 11052340 TI - A profile of dental school deans. PMID- 11052339 TI - Role of capsaicin-sensitive nerves in gastric and hepatic injury induced by cold restraint stress. AB - The role of capsaicin-sensitive afferent fibers on cold-restraint stress-induced gastric and hepatic injury was examined at the macroscopic and ultrastructural levels. Wistar albino rats were treated with capsaicin either locally (intragastric, perivagal, and periceliac) or systemically (neonatal, intraperitoneal). Perineural and neonatal treatment with capsaicin was used to denervate afferent fibers, while intragastric capsaicin treatment would have activated mucosal afferent fibers just before the stress exposure. Capsaicin decreased significantly the formation of macroscopic gastric lesions caused by stress in all treatment groups. At the electron microscopic level, however, denervation of vagal afferent fibers with capsaicin was most effective in prevention of cellular injury in gastric mucosa. In the liver, systemic denervation of afferent fibers completely inhibited stress-induced cellular damage, while denervation of afferent fibers in vagus and splanchnic nerve was partially effective. Central neural pathways sensitive to capsaicin may mediate formation of both gastric and hepatic injury resulting from stress. PMID- 11052341 TI - Dental students' attitudes toward smoking cessation guidelines. AB - Dentists can be effective in helping their patients achieve smoking cessation. To plan a didactic program, we explored the smoking cessation attitudes and practices of dental students and identified barriers to service provision in the dental setting. We assessed 244 fourth-year dental students at New York University College of Dentistry through a self-report survey. The instrument included a twenty-nine-item measure assessing attitudes towards tobacco-use counseling and adherence to National Cancer Institute tobacco cessation guidelines. The survey also assessed demographics, tobacco use history, and level of preparation to provide services. Generally, students endorsed tobacco prevention practices, but perceived barriers to service provision. Students provided counseling inconsistently, with 69 percent asking about smoking, 58 percent advising cessation, 24 percent offering assistance, and 22 percent providing followup on a routine basis. Those who provided more counseling were more likely to have undergone formal training in smoking cessation, did not feel time was a barrier to counseling, and had more favorable beliefs about dentists' role in promoting smoking cessation. Study findings indicate great receptivity among students as well as a critical need and opportunity to include comprehensive cessation counseling training in the dental curriculum. PMID- 11052342 TI - An initial evaluation of standards for case presentations. PMID- 11052343 TI - Dental school faculty shortages increase: an update on future dental school faculty. AB - The 1999 publication of the American Association of Dental Schools (AADS) President s Task Force on Future Dental School Faculty revealed a crisis in the shortages of dental school faculty. Stakeholders from around the nation have used the AADS Task Force report to address the crisis. In addressing one of the AADS Task Force recommendations, the American Dental Education Association (ADEA), formerly AADS, gathered additional data through a new survey of dental school deans to elucidate the current state of faculty shortages. Based on this research, ADEA projects that the number of unfilled budgeted faculty positions in U.S. dental schools now approaches 400. Survey respondents identified retirement as the leading reason for full-time faculty separations, while separation to enter private practice was the second most frequent reason for leaving the institution. Offering a salary competitive with that of private practice was identified as the most critical factor in recruiting future faculty. A number of short and long-range strategies to recruit and retain faculty are presented. Ultimately, the dental school faculty shortage places in jeopardy the general and oral health of the public. PMID- 11052344 TI - A prospective comparison of the standard and reverse robinson cervical grafting techniques: radiographic and clinical analyses. AB - The authors performed a prospective study of 63 patients with cervical radiculopathy treated with Robinson anterior cervical discectomy and fusion and compared the traditional or standard and reverse graft techniques. Modifications of the standard Robinson grafting techniques have been proposed. The reverse graft technique has theoretical advantages, including limiting the deleterious effects of graft extrusion and maintaining rigid middle column support. A radiographic evaluation and an assessment of clinical outcome based on the criteria of Odom were performed prospectively for as long as 1 year after surgery. Thirty-one patients were treated with the standard grafting technique and 32 with the reverse graft orientation. The radiographic evaluation showed no significant differences between the two techniques with regard to sagittal alignment and disk heights. The overall fusion grade was higher in the reverse graft technique (p < 0.05). There were 93% and 96% good to excellent results in the standard graft and reverse graft groups, respectively. The authors report no significant differences associated with the standard or reverse anterior cervical grafting techniques in terms of radiographic alignment or disk height loss over time or at early clinical outcome. However, improved fusion grade was noted with the reverse graft technique, which may be related to end plate and intervertebral space preparation. The reverse grafting technique is an acceptable alternative to the standard graft orientation. PMID- 11052345 TI - The management of traumatic cervical bilateral facet fracture-dislocations with unicortical anterior plates. AB - The goal of this study was to evaluate single-level anterior cervical discectomy and stabilization for bilateral facet fracture dislocations using bone graft and anterior titanium plates with unicortical screw fixation in the clinical setting. We conducted a retrospective review during a consecutive 6-year period of patients treated in a single institution for traumatic single-level cervical bilateral facet fracture-dislocation. All fracture-dislocations that could be aligned with traction were subsequently stabilized using an anterior cervical discectomy with bone graft and titanium unicortical locking plates. All patients were examined after operation for radiographic evidence of healing and neurologic outcome. Twenty-two patients (M:F ratio, 16:6; average age, 47.7 years) underwent an anterior cervical discectomy and stabilization with either an allograft (n = 12) or an autograft (n = 10) and a titanium unicortical locking plate. Most patients had sustained a spinal cord injury (87%) or a radicular injury (9%). The average follow-up was 32 months, with a minimum follow-up of 1 year (range, 13 to 77 months). There was one instrumentation-related failure, but all 22 patients ultimately had evidence of stability at the instrumented level on the final follow-up examination. Anterior fixation with unicortical cervical locking plates as a single procedure offers an excellent surgical alternative in the management of many cervical bilateral facet fracture dislocations in patients who can be reduced preoperatively. PMID- 11052346 TI - Role of onlay grafting with minimal internal fixation for occipitocervical fusion in oncologic patients. AB - The choice of fixation for occipitocervical arthrodesis remains controversial, especially in oncologic patients who need further radiographic evaluation or postoperative radiotherapy. We studied the long-term outcome of 20 patients who underwent occipitocervical fusions using onlay corticocancellous bone graft and wiring, with postoperative halo vest immobilization. Eight of these patients had postoperative radiotherapy for spinal tumors (group I), and 12 patients had occipitocervical fusions for other pathologies that did not require postoperative radiotherapy (group II). All patients had solid arthrodeses; however, there was a difference in the average time to fusion between patients who received pre- or postoperative radiotherapy and those who did not (p = 0.11). At an average follow up of 54.5 months (range 24-92 months) 15 of 20 patients (75%) had excellent or good outcomes. A high fusion rate can be expected with reasonable intraoperative or postoperative complications. PMID- 11052347 TI - Effects of dexamethasone and of local hypothermia on early and late tissue electrolyte changes in experimental spinal cord injury. AB - The current experiment reexamines this laboratory's frequently cited previous experimental conclusion that a mechanism underlying the beneficial effects of glucocorticoids in the treatment of spinal cord injury may be the enhanced preservation of spinal cord tissue potassium. For the first time, similar methodology also has been applied to study the effects of hypothermia. Canine spinal cords were injured at T13 by use of an epidural balloon and then were treated with local hypothermia or intramuscular dexamethasone or both. Motor recovery was assessed using a modified Tarlov scale. At either 6 days or 7 weeks, spinal cords T8 through L4 were removed and divided into 10 ordered blocks, which were analyzed for wet and dry weight, potassium concentration, and sodium concentration. Correlations between clinical motor and chemical results were evaluated. The conclusions drawn are as follows: 1) The canine severe rapid compressive injury model, unlike the previously published less severe feline impact injury model, is not associated with widespread early loss of spinal cord tissue potassium content (dry weight). 2) The dog compressive model, unlike the cat impact model, does not provide evidence that one fundamental mechanism of the confirmed beneficial action of steroids entails enhanced early preservation of tissue potassium content. 3) At 6 days, decrease in the percentage of dry weight and increase in sodium concentration, representing edema, occurred at and adjacent to the direct compression site in all lesioned dog groups except those treated with dexamethasone, demonstrating an antiedema effect of dexamethasone that was nullified by concurrent local hypothermia. 4) This antiedema effect of dexamethasone was associated with superior early motor improvement but did not lead to superior long-term function, in comparison with hypothermia. 5) At 7 weeks, decrease in the percentage of dry weight and potassium concentration, and increase in sodium concentration, all restricted to the directly compressed segment, signify necrosis. 6) This new chemical index of necrosis was highly correlated with clinical motor performance. PMID- 11052348 TI - Anterior dural laceration caused by thoracolumbar and lumbar burst fractures. AB - A retrospective review of the records of 60 patients with thoracolumbar and lumbar burst fractures was undertaken to document the incidence and evaluate the sequelae of dural injuries found during anterior procedures. In the entire series, six (10%) patients each had a preexisting vertically oriented dural tear. All patients with anterior dural lacerations were male and had associated neurologic deficits. In all six patients, preoperative computed tomography showed an asymmetrically retropulsed bone fragment. Dural tears were repaired primarily. A postoperative cerebrospinal fluid leak developed into the chest cavity of one patient, who was treated successfully with subarachnoid drainage. In patients with anterior dural laceration, primary repair is warranted and can be performed more easily after intraoperative correction of kyphosis. Subarachnoid drainage may be effective in cases of continued postoperative anterior cerebrospinal fluid leakage before repeated operation is considered. PMID- 11052349 TI - Radiographic parameters for evaluating the neurological spaces in experimental thoracolumbar burst fractures. AB - It is important to know the condition of neural spaces during the nonoperative treatment of thoracolumbar burst fractures. The goals of the current study were to identify the correlation between the degree of deformity of a fractured vertebra and the encroachment of neural spaces, and to determine how the encroachment and the deformity can be improved by the extension posture simulating the postural reduction. Experimental burst fractures were produced in L1 vertebrae of nine human thoracolumbar spine segments (T11-L3) with neural spaces lined with tiny steel balls. Lateral radiographs were taken in neutral and extended posture before and after the trauma. Anterior vertebral height, posterior vertebral height, vertebral height ratio, vertebral kyphotic angle, posterior vertebral body angle, and the cross diagonal angle were the geometric parameters used to describe the vertebral deformity. The canal diameter and superior and inferior intervertebral foramen areas were defined as the neural spaces. All parameters were measured on the scanned images of radiographs, as seen on the computer screen. Among the vertebral body parameters, the posterior vertebral height, posterior vertebral body angle, and cross diagonal angle showed significantly higher correlations with the canal encroachment. The extended posture did not improve the canal and intervertebral foramen encroachments. The kyphotic deformity (vertebral kyphotic angle and anterior vertebral height) was improved but the deformity of the vertebral posterior wall (posterior vertebral height and posterior vertebral body angle) was not improved because of the extended posture. PMID- 11052350 TI - Accuracy of fluoroscopically assisted laser targeting of the cadaveric thoracic and lumbar spine to place transpedicular screws. AB - A simple and inexpensive method was developed to obtain a coaxial view of the pedicles to assist with screw insertion. The authors evaluated the accuracy of this device to place transpedicular vertebral screws in a human adult cadaver model. A dual radiation targeting system, a laser targeting system for fluoroscopically guided procedures, was developed to provide an accurate surface entry point and angle of approach to radiographic landmarks. After fluoroscopic cross-hair target localization of the coaxial view of the pedicle, X-ray radiation is turned off and the laser beam allows the surgeon to guide the screw through the pedicle. Nine cadaver spines were removed and mounted. Three surgeons, inexperienced in the technique of pedicle screw placement, fitted instruments to 184 pedicles between L5 and T5. A total of 83 lumbar and 101 thoracic pedicles underwent screw placement. After specimen dissection, the degree and location of any screw perforation were measured by direct inspection. Three screws perforated a pedicle, for an error rate of 1.6%. Two lumbar screws (2.4% error) and one thoracic screw (1% error) perforated the pedicle. No screw was more than 1 mm outside the pedicle. Five other screws, four in the thoracic spine and one in the lumbar spine (error rate of 2.7%) were directed too far laterally and perforated the lateral vertebral body. This low rate of pedicle wall cortical perforation by inexperienced surgeons compares favorably with much higher pedicle perforation rates by experienced surgeons when no imaging was used. In conclusion, this in vitro model using a dual radiation targeting system assisted with the accurate placement of transpedicular vertebral screws with minimal radiation exposure. PMID- 11052351 TI - Internet resources for spine surgeons. AB - To classify web sites on common spinal disorders as to their utility for the spine surgeon and patient. Five common spinal disorders were used to generate lists of relevant sites. These sites were categorized as to their relevance for patients and surgeons, their sponsoring organization, and their comprehensiveness. A total of 56,249 web sites were found using the five key words on five search engines. Using the "And" operator, a total of 227 web sites were generated. The majority of sites were patient oriented. Physician- or organization-sponsored sites were the most common. Ten sites were found to have comprehensive information for both patients and spine surgeons. Many web sites exist that discuss disorders of the spine. Currently there is not any one web site that contains comprehensive information for both the spine surgeon and patient. PMID- 11052352 TI - Postoperative spinal wound infection: a review of 2,391 consecutive index procedures. AB - Postoperative infection remains a troublesome but not uncommon complication after spinal surgery. Most previous reports, however, are small or involve cases with more than one surgeon often at different institutions. This study represents a single surgeon's 9-year experience with postoperative infection at one institution. The authors describe the features of wound infection after spinal surgery with reference to diagnosis, microbiology, and treatment and they describe a protocol for effective management of postoperative spinal wound infection. The records of the senior author (F.P.C.) during a 9-year period for cases of postoperative wound infection were reviewed. Of 2,391 operative procedures, 46 cases of wound infection were identified, yielding an overall infection rate of 1.9%. Patients' preoperative risk factors, original diagnosis prompting the surgery, onset of infection, presentation, treatment, and outcome were analyzed. The mean age of the 23 men and 23 women was 57.2 years. The preoperative diagnoses included lumbar degenerative scoliosis or spinal stenosis in 28 cases, disk prolapse in 8 cases, metastatic disease in 4 cases, degenerative disk disease in 1 case, and a group of 5 miscellaneous cases. Seventeen (37%) of the patients underwent at least one previous spinal surgery at the same site. Twenty-three patients had a fusion, of whom 22 also had instrumentation. Forty-three (93%) of the patients had significant wound drainage after an average of 15 days (range, 5-80 days). The other three patients were examined approximately 2 years after the surgery. Fourteen of the patients also had pyrexia (temperature >37.5 degrees C) at presentation. Staphylococcus aureus alone was cultured in 29 patients, whereas another six patients had a different single organism. In nine patients, more than one organism was cultured during their hospital stay. Surgical treatment included primary closure in only seven patients, with most undergoing wound drainage and debridement followed by delayed closure. Instruments were removed in the three patients with late presentation who had solid fusion at operation. Viable bone graft and instrumentation were left in situ in all patients who were seen before fusion. All wounds healed without sequelae, except for three that required flap closure. Pseudarthrosis was noted in three patients after more than 1 year of follow-up in this series. Postoperative spinal wound infection is a potentially devastating problem. In this series, infection was more common in patients undergoing fusion with instrumentation and in patients with cancer metastatic to the spine. An aggressive surgical approach, including repeated debridement followed by delayed closure, is justified. Instrumentation may be safely left in situ to provide stability for fusion. PMID- 11052353 TI - Recovery of function in adjacent nerve roots after surgery for lumbar disc herniation: use of quantitative sensory testing in the exploration of different populations of nerve fibers. AB - Results from experimental and clinical studies indicate that adjacent nerve roots may be affected in sciatica because of lumbar disc hemiation. This may be caused by proinflammatory mediators in the epidural space being transported into nerve roots at the same or neighboring lumbar segments. The aim of the present study was to investigate the recovery of function of sensory nerve fibers in the adjacent noncompressed nerve roots. Thirty-nine patients undergoing microdiscectomy for monoradiculopathy were investigated with quantitative sensory testing immediately before surgery, and at 6 weeks, 4 months, and 12 months after the operation. Twenty-one healthy volunteers were used as controls. The patients were classified as having a good or a poor result at the 1-year follow-up according to a clinical score. Significant improvement of function in the noncompressed nerve roots were only observed in the 31 patients with a good result. The improvement in small myelinated nerve fibers came within 12 months in the adjacent nerve roots in both the symptomatic as well as the asymptomatic leg. The improvement of function in small unmyelinated fibers also came within 12 months after surgery; however, significant improvement was only observed in the ipsilateral neighboring nerve root. The function in large myelinated fibers did not improve in any of the adjacent nerve roots during the observation period. The observed recovery of function in adjacent noncompressed nerve roots after successful surgical decompression in monoradiculopathy may be because of less production of proinflammatory mediators when the disc herniation is removed. PMID- 11052354 TI - Results of surgery for spinal stenosis adjacent to previous lumbar fusion. AB - The literature provides little data to guide surgical management of spinal stenosis adjacent to previous lumbar fusion. Thirty-three consecutive patients who had surgical decompression for spinal stenosis at the lumbar segments adjacent to a previous lumbar fusion were studied. The mean interval between fusion and the adjacent segment surgery was 94 months. Of the 33 patients, 26 were followed for 3-14 years (mean: 5 years) after adjacent segment surgery and were clinically evaluated and independently completed an outcome questionnaire. Of the 26 patients, 15 rated their outcome as completely satisfactory, 6 were neutral toward the surgery, and 5 considered their surgery a failure. The surgery was generally effective at improving or relieving lower extremity neurogenic claudication. The strongest independent predictive factor of patient dissatisfaction was ongoing postoperative low back pain (r = 0.7, p = 0.001). A higher back pain score at follow-up was associated with continued narcotic use (p = 0.001) and decreased ability to perform activities of daily living (p = 0.05). Six patients required further lumbar surgery during the follow-up period. This study provides the longest published follow-up data of surgical results for symptomatic spinal stenosis adjacent to a previously asymptomatic lumbar fusion. PMID- 11052355 TI - Association between findings of provocative discography and vertebral endplate signal changes as seen on MRI. AB - Provocative discography is a controversial diagnostic tool for pathologic discs. Modic has identified vertebral endplate signal changes on magnetic resonance imaging (MRI) that are thought to signify advanced discogenic degeneration. These two distinct diagnostic tools are examined to determine if there is association between them. Fifty-three consecutive patients who underwent both investigations were retrospectively reviewed. In discs that had negative T1 MRI findings, 28.2% of patients had concordant pain and 17.3% had discordant pain. In discs with positive T1 MRI findings, 34.8% of patients had concordant pain and 17.4% had discordant pain. 79.5% and 74.4% of levels with patient concordant pain on discography had no endplate changes on T1- and T2 weighted MR images, respectively (compared with 84.5% and 81.7%, respectively, for levels with no patient pain on discography). Our data showed no significant relationship between these distinct diagnostic tools. Further investigation of their relative roles in this application is recommended. PMID- 11052356 TI - The relationship between disc degeneration, facet joint osteoarthritis, and stability of the degenerative lumbar spine. AB - Degenerative processes in the disc and facet joints affect the stability of the motion segment. The exact relations among disc degeneration, facet joint osteoarthritis, and the kinematics of the motion segment are not well defined in the literature. Magnetic resonance imaging and functional radiography of the lumbar spine were analyzed to examine the relations among segmental instability, facet joint osteoarthritis, and disc degeneration in patients with degenerative disorders of the lumbar spine. Seventy consecutive patients (mean age, 46 years) had both magnetic resonance imaging and flexion and extension radiographs of the lumbar spine. The lumbar instability was classified into abnormal tilting on flexion, rotatory instability in the sagittal plane, and translatory instability. Translatory instability was subdivided into anterior, posterior, and anteroposterior translatory instability. Disc degeneration as seen on T2-weighted sagittal images was classified into five grades. Facet joint osteoarthritis as seen on axial T1-weighted images was divided into four grades. This study revealed that the kinematics of the lumbar motion segment are affected by disc degeneration and facet joint osteoarthritis. Abnormal tilting movement on flexion and anteroposterior translatory instability both had negative associations with facet joint osteoarthritis. However, anterior translatory instability was positively associated with disc degeneration and facet joint osteoarthritis. Rotatory instability in the sagittal plane and posterior translatory instability were not associated with disc degeneration and facet joint osteoarthritis. PMID- 11052357 TI - A rare cause of lumbar radiculopathy: spinal gas collection. AB - The presence of gas in the intervertebral disk space, known as the vacuum phenomenon, is a relatively common radiologic finding, especially on computed tomographic investigation. In a few cases, the gas can be collected into the lumbar spinal canal and can also compress the nerve root. To date only seven cases of symptomatic lumbar radiculopathy caused by a bubble of gas are reported in the literature. The presence of gas inside a narrowed disk and the collection of gas in the spinal canal suggest a communication between the two structures. A case of lumbar radiculopathy caused by a collection of gas in the spinal canal provided the authors the opportunity to study this rare condition by magnetic resonance imaging. Magnetic resonance imaging had not been used before in the referred cases and proved conclusively the discal origin of the gas. PMID- 11052358 TI - Isolated thoracolumbar and lumbar hyperlordosis in a patient with cerebral palsy. AB - A severe isolated thoracolumbar and lumbar hyperlordosis spinal deformity occurring in a patient with cerebral palsy is rare and has not been reported before. The authors describe the presentation, operative considerations, and treatment of patients with this unusual hyperlordotic spinal deformity, particularly those with cerebral palsy. A multiple-stage surgical reconstruction was required to correct this complex spinal deformity. The patient underwent bilateral femoral extension osteotomies along with spinal extensor myotomies to ensure proper prone positioning for his anticipated spinal surgery. Then he had staged anterior releases and spinal fusion from T8 to the sacrum followed by 2 weeks of "90-90" femoral skeletal traction. Finally, a posterior spinal fusion with instrumentation from T2 to the pelvis definitively corrected his deformity. The patient responded well to surgical intervention without complications and continues to have stable correction of his hyperlordosis deformity 2 years after surgery. Severe lordotic sagittal plane spinal deformities can be treated with anterior and posterior spinal fusion and instrumentation with intervening traction in the properly selected and prepared patient who has cerebral palsy. PMID- 11052359 TI - An approach to student learning in clinical radiology. PMID- 11052360 TI - The computed tomographic appearance of acute thoracolumbar intervertebral disc herniations in dogs. AB - The appearance of herniated intervertebral disc material in the thoracolumbar vertebral canal was evaluated in 23 dogs using computed tomography (CT). The images were then compared with the myelographic and surgical findings. The normal spinal cord, outlined by epidural fat over intervertebral disc spaces, was of intermediate attenuation on transverse CT images. Herniated disc material was identified in all animals as a heterogeneous hyperattenuating extradural mass. The attenuation of the disc material increased with the degree of mineralization. In seven dogs, the herniated material was only slightly more attenuating than the spinal cord. In these dogs, small fragments of mineralized disc material and significant hemorrhage were found in the epidural space at surgery. In dogs with a long standing history of disc herniations, disc material identified in the vertebral canal had a more hyperattenuating and homogeneous appearance than recently herniated disc material. We conclude that mineralized, herniated disc material and hemorrhage can be identified quickly and safely in dogs using CT. PMID- 11052361 TI - Dynamic CT measurement of contrast medium washin kinetics in canine nasal tumors. AB - Tumor oxygenation affects the biologic behavior of a tumor and also its radiation response. Decreased tumor oxygenation has been associated with an aggressive phenotype and with decreased local tumor control following irradiation. Thus, measurement of oxygenation may be useful for pretreatment evaluation of a tumor. Many methods for assessing tumor oxygenation are available but most are invasive. There is a need for a non-invasive measure of oxygenation, or a surrogate for oxygenation. Measurement of perfusion has been suggested as a substitute for measurement of oxygenation. The use of washin kinetics of iodinated contrast medium to estimate perfusion has been shown to be related to radiation response of human carcinomas. We quantified the washin kinetics of iodinated contrast medium using dynamic CT in 9 dogs. All dogs had a malignant nasal tumor and perfusion was quantified at two sites in each tumor to evaluate intratumoral variation in perfusion. Dogs were given an intravenous bolus injection of contrast medium and arterial and tumor washin kinetics quantified using a helical CT scanner. Perfusion was estimated from these data using previously validated methods. Eight of the 9 dogs received definitive radiation therapy and perfusion was quantified a second time in these 8 dogs midway through irradiation. Pretreatment perfusion varied between dogs by a factor of 16.9. Between dog variation in perfusion was subjectively greater than within tumor variation based on comparison of two intratumoral regions. Changes in perfusion in individual dogs during irradiation were observed, but no identifiable pattern of perfusion alteration was detected. Measurement of perfusion in canine nasal tumors using dynamic CT is possible and further study of this parameter as it relates to radiation response is reasonable. PMID- 11052362 TI - Quantitative videofluoroscopic evaluation of pharyngeal function in the dog. AB - Videofluoroscopic evaluation of both liquid barium and barium soaked kibble was performed in 11 adult, clinically normal dogs of varying breeds. Each examination was digitized and evaluated frame by frame to establish the normal timing sequence of the pharyngeal phase of swallowing. Closure of the epiglottis was considered the onset of swallowing. The time to each of the following events was recorded: (1) Maximum pharyngeal contraction, (2) Opening of the epiglottis, (3) Opening of the cranial esophageal sphincter, and (4) Closure of the cranial esophageal sphincter. These values were found to be consistent both intra and interpatient. Retrospective analysis of 3 videofluoroscopic examinations from dogs that met the subjective criteria defining cricopharyngeal achalasia was then performed. A statistically significant delay in the time to opening and closure of the cranial esophageal sphincter was found employing both liquid barium and barium soaked kibble in the dogs with cricopharyngeal achalasia. PMID- 11052363 TI - Magnetic resonance imaging of the normal and diseased feline middle ear. AB - The magnetic resonance imaging appearance of the feline middle ear is described in three healthy cats and in five cats with middle ear disease. Owing to the good spatial resolution, multiplanar slice orientation as well as display high contrast resolution of soft tissue, in particular fluids, MR imaging was helpful prior to surgery. It is superior to radiography which failed to allow identification of the abnormality in two of our five cats. MR imaging for middle ear disease should include dorsal and transverse plane images using T1- and T2 weighted sequences. In the presence of a mass within the bulla or the external ear canal application of contrast medium is helpful. PMID- 11052364 TI - Radiographic diagnosis: aural abscesses in a box turtle. PMID- 11052365 TI - Bronchocutaneous fistula in a dog. AB - A dog with a bronchocutaneous fistula is described. Contrast enhanced computed tomography was useful for diagnosis and treatment of the fistula in this dog. PMID- 11052366 TI - Post-operative computed tomography in two dogs with cerebral meningioma. AB - Post-operative computed tomography (CT) has been described as a technique for diagnosing incomplete resection or recurrence of cerebral neoplasms in humans. The characteristics of immediate postoperative CT images in dogs with intracranial pathology are unknown. This report describes findings from preoperative, immediate post-operative, and 4 week to 9 month follow-up CT examinations in two dogs with histologically-confirmed cerebral meningiomas. In images of one dog after surgery there was mild contrast enhancement of the tissue surrounding the surgical site. This enhancement had resolved in later images and was probably the result of surgically induced trauma. In post operative images of the other dog there was significant hyperattenuation of the tissues around the surgical site. In post contrast images there was increased enhancement that was evident in later images. These findings, although not supported by necropsy, probably indicate incomplete excision of the tumor. PMID- 11052367 TI - Alternatives for improving veterinary medical students' learning of clinical sonography. AB - With the widespread clinical use of sonography there is a need to introduce the topic into the curriculum. A new problem-based course in clinical sonography without lectures was developed to emphasise experiential learning, and engage students actively in individual and collective acts of discovery. Four different approaches were used to deliver the new course to 141 veterinary medical students over four semesters. The physical principles of sonography were taught by computer-assisted instruction and a practical class, clinical examinations were introduced during a session with a tutor, and finally each student wrote an essay on a sonographic topic of their choice. To evaluate the new course, students' responses to a questionnaire were analyzed. Students gained reasonable understanding of the physical principles of sonography and had some confidence in conducting a sonographic examination of an animal. Of most use to student learning was discussion with the teachers. Surprisingly, half the students thought the topic should also be taught by lectures. The students learned the material and acquired the sonographic skills through processes which required more independence and self-responsibility than traditional teaching methods. The teachers' interaction with students on an individual basis, as they encountered individual problems, was the most important resource in learning about sonography. The continued request for lectures suggests an insecurity in some students caught between two different paradigms of teaching and learning (experiential, problem-based learning versus lectures). PMID- 11052368 TI - Relevance of sonographic artifacts observed during in vitro characterization of urocystolith mineral composition. AB - Nine pure mineral type canine uroliths (bladder or urethral origin only) were imaged ultrasonographically using 3.5 MHz, 5.0 MHz, and 7.5 MHz fixed focus, mechanical sector transducers in a urinary bladder phantom. The uroliths studied were those composed of 100% magnesium ammonium phosphate, calcium oxalate monohydrate, calcium oxalate dihydrate, calcium phosphate appatite, and calcium hydrogen phosphate dihydrate (brushite), ammonium acid urate, sodium acid urate, cystine, and silica. The occurrence of both reverberation/streak and acoustic shadowing artifacts were compared to urocystolith mineral type (classified by effective atomic number), urocystolith width, urocystolith height (thickness), and ultrasonographic imaging frequency. No predictable relationship was found between either of the artifacts seen beyond the urocystolith (reverberation/streak or the acoustic shadowing) and urocystolith mineral type. There was no statistical relationship between the occurrence of reverberation/streak artifact and the size (width or height) of the urocystolith or the ultrasonographic frequency. There was, however, a statistically relevant relationship between ultrasonographic imaging frequency and the occurrence of acoustic shadowing and between urocystolith height (thickness) and the occurrence of acoustic shadowing. However, regardless of ultrasound frequency, acoustic shadowing was observed less than 35% of the time in any of the urocystolith mineral types examined. Based on the imaging of the bladder phantom supporting apparatus (7.0 mm bolts covered by plastic), the accurate characterization of a curved object surface directly facing the transducer was found to be directly related to the frequency of sound used for imaging and at best predictably limited to curved vs flat. Accurate measurement of the maximum transverse dimension of an echogenic curved object or accurate characterization of the lateral borders of such an object was considered unlikely with general ultrasonographic equipment of the frequencies studied. Therefore, detailed architectural characterization of urocystoliths suitable for mineral composition prediction is considered highly unlikely with general pulse-echo ultrasonographic techniques. PMID- 11052369 TI - Ultrasound evaluation of the parathyroid glands in two hypercalcemic cats. AB - Two parathyroid masses were identified using ultrasound in two hypercalcemic cats. The masses were identified as parathyroid adenomas on surgical biopsy. Both parathyroid masses contained hypoechoic regions with distal acoustic enhancement. Both masses were greater than 1.0 cm in diameter. PMID- 11052370 TI - M-mode and Doppler echocardiographic findings in normal ferrets sedated with ketamine hydrochloride and midazolam. AB - M-mode and Doppler echocardiographic values were obtained from 30 normal adult ferrets (14 neutered females, 13 neutered males, 3 intact males) sedated with an intramuscular combination of ketamine hydrochloride and midazolam. Routine M-mode measurements of the left and right ventricle, left atrium (LA) and aorta (AO) and Doppler measurements of aortic and pulmonic outflow, and mitral inflow were recorded. The following values were calculated: LA:AO diameter, ratio of peak E: peak A wave velocity (E:A ratio) for mitral inflow, and stroke volume (SV), cardiac output (CO) and cardiac index (CI) for both pulmonary and aortic outflow tracts. Maximal aortic velocities (AOmax) and velocity-time integral values (AO VTI) were significantly less than corresponding pulmonary outflow tract values (PAmax, PA VTI) but there was no difference in calculated values for SV, CO or CI. Calculated CO values were in the range expected based on the size of the species. Difficulties in aligning the aortic outflow tract for Doppler imaging may make pulmonary outflow Doppler values more consistent for use in estimating volume flow in ferrets. PMID- 11052371 TI - Ultrasonographic evaluation of tarsocrural joint cartilage in normal adult horses. AB - Ultrasonographic examination of the tarsus was performed on four clinically and radiographically normal limbs of adult horses. Particular attention was paid to the articular cartilage surfaces of the trochlear ridges of the talus and the distal intermediate ridge of the tibia. Two separate measurements of articular cartilage thickness were acquired from a longitudinal view at each site. Anatomy was confirmed with post mortem dissection. Ultrasonography was found to be a practical method for imaging the articular cartilage over the trochlear ridges of the talus and distal intermediate ridge of the tibia. The cartilage appeared as a hypoechoic band overlying the hyperechoic subchondral bone. The mean cartilage thickness over the lateral and medial trochlear ridges of the talus and the distal intermediate ridge of the tibia were 0.57 mm, 0.58 mm and 0.7 mm respectively. These measurements may have value for comparison to thickened cartilage and lesions of osteochondrosis and abnormally thinned cartilage of osteoarthritis. Ultrasound examination was not helpful in evaluating the proximal and distal intertarsal and tarsometatarsal joints, the close proximity of the articular surfaces obscured visualization of the articular cartilage. PMID- 11052372 TI - Double-phase parathyroid scintigraphy in dogs using technetium-99M-sestamibi. AB - The purpose of this study was to evaluate the utility of double-phase parathyroid scintigraphy using 99mTc-sestamibi for detecting and localizing hyperfunctioning parathyroid glands in hypercalcemic dogs. Fifteen hypercalcemic dogs that underwent parathyroid scintigraphy were included in this study: 3 dogs with hypercalcemia of malignancy, and 12 dogs with hyperfunctioning parathyroid tissue (parathyroid adenoma or parathyroid hyperplasia). The presence of parathyroid adenoma or parathyroid hyperplasia was documented by histopathologic examination. In 3 dogs with hypercalcemia of malignancy, parathyroid scintigraphy was negative for hyperfunctioning parathyroid tissue and the scans were classified as true negative. Parathyroid scintigraphy correctly identified the presence and location of hyperfunctioning parathyroid tissue in only 1 of 6 dogs with a parathyroid adenoma. False positive and false negative results occurred in dogs with parathyroid adenomas. Parathyroid scintigraphy failed to detect hyperfunctioning parathyroid tissue in 5 of 6 dogs with parathyroid hyperplasia and were classified as false negative. False positive results were obtained in the remaining dog with parathyroid hyperplasia. Sensitivity of parathyroid scintigraphy for detecting and localizing hyperfunctioning parathyroid tissue was 11%, specificity was 50%, and overall accuracy was 27%. Positive and negative predictive value were 25% and 27%, respectively. Sensitivity for detection of parathyroid adenomas was 25%, and sensitivity for detection of hyperplastic glands was 0 %. Results of this study indicate that double-phase parathyroid scintigraphy does not appear to have acceptable accuracy in detecting hyperfunctioning parathyroid glands in dogs. Due to the poor sensitivity and specificity of the technique in dogs, parathyroid scintigraphy is not recommended for definitive identification of abnormal parathyroid glands as the cause of hypercalcemia in dogs. PMID- 11052373 TI - Ancillary equipment to increase quality and reduce radiation exposure in the equine nuclear medicine laboratory. AB - The construction of ancillary equipment used to improve image quality and reduce personnel radiation exposure in the equine nuclear medicine laboratory is illustrated. The devices include a self supporting lead sheet for shielding the distal limb or limb pair, a hanging lead sheet for shielding the proximal limb, a lead square for shielding the urinary bladder or jugular catheter, a restraining board for acquiring a palmar view of the foot, a head support to stabilize the head for imaging and a head support for stabilizing the neck for imaging. The restraining board and head supports decrease patient motion, improve image quality, and decrease the number of repeat acquisitions so that the time to perform the complete study is shortened. The self supporting shielding device allows the handlers to increase their distance from the horse which enhances radiation safety. PMID- 11052374 TI - Prologue: discourse concerning two conventional sciences. AB - The form of the Discorsi of Galileo Galilei is used here to emphasize the three aspects of research objects. The point of view of the user (Simplicio), the theoretician (Salviati) and the experimenter (Sagredo). Two technical aspects of the papers which have been presented in the three Biomechanics Supplements of Injury [volumes 30(1), 31(2), and 31(3)] are discussed by these three interlocutors: i. The confidence intervals of the number of cases in the epidemiological studies, which should be high enough to reveal significant differences (i.e. a total of about 2,000 patients to show that an improvement in treatment reduces the infection rate); ii. The importance of the deformation for the quantification of the deleterious effect of stress protection, whereby it is shown that the size of the subject (small animal vs human) is non-linearly related to the effect, as was already disclosed by Galilei 367 years ago--but which is still not yet well understood by the nonmechanic. PMID- 11052375 TI - Stress protection due to plates: myth or reality? A parametric analysis made using the composite beam theory. AB - A generally accepted idea has been that plate fixation of fractures may result in the structural adaptation of bone (bone loss) to reduced stress (stress protection) with the subsequent danger of refracture after implant removal. This was the negative aspect of stress protection. For this reason, it was proposed that plates made from more deformable materials be used (titanium, polymers or carbon fibres). A theoretical analysis using composite beam theory, with different loading conditions (axial load and bending), demonstrates that stress protection, i.e. early temporary porosis, is a myth. Mechanics of materials shows that when an over-large plate is fixed to small bones (as in small animals, e.g. rabbits), the reduction of bone strain is exaggerated; in contrast, using plates of varying flexibility (steel, titanium or carbon fibre) on large bones leads to strain reduction with an astonishingly similar amplitude. PMID- 11052376 TI - Effect of plate position relative to bending direction on the rigidity of a plate osteosynthesis. A theoretical analysis. AB - Mechanical unloading of the plated bone segment is observed after plate osteosynthesis because the implant takes over a part of the physiological loading. Strain reduction in the bony tissue depends on the rigidity of the plate (cross-sectional area, geometrical form, and modulus of elasticity). The aim of the present study was to calculate theoretically the effect of plate position relative to bending direction on the overall bending stiffness of the composite system plate-bone. To calculate the rigidity, a cylindrical bone model with mechanical characteristics similar to a sheep tibia and a rectangular plate cross section corresponding to a DC-plate with either a modulus of elasticity of steel or titanium was used. Calculations under different bending directions were performed according to the laws of the linear bending theory and the composite beam theory. The bending stiffness of a plate osteosynthesis reaches a minimum and a maximum respectively, in cases in which the bending moment acts in the direction of the main axis of the area moment of inertia of the plate. The minimum is present with the plate bent vertically, the maximum with the plate bent horizontally, e.g. on the tension side of the composite system--on the assumption that the bone structure opposite the plate is capable of withstanding compressive loading. For steel and titanium plates, factors of 2 and 2.25 respectively were calculated between the minimum and the maximum bending stiffnesses of the osteosynthesis. The bending rigidity of the plate alone has only a minimal effect on the total stiffness of the osteosynthesis. With a plate bent vertically, the difference between steel and titanium plates was 18%, with the plate bent horizontally (situated on the tension side), it was only 7%. The bending stiffness of a plate osteosynthesis depends on the cross-section, the geometrical form, and the modulus of elasticity of the plate, as well as on the plate position relative to the bending direction of the composite system. The modulus of elasticity of the plate is relatively unimportant, while with a given plate the individual plate position relative to the bending direction is of crucial importance. Thus, changing the modulus of elasticity of the plate cannot solve the problem of implant induced unloading of the bone cortex because the bending stiffness of the composite system depends much more on the plate position relative to the bending direction. PMID- 11052377 TI - Force transfer between the plate and the bone: relative importance of the bending stiffness of the screws friction between plate and bone. AB - Stability in plate fixation of fractures relates to the motionless fastening between plate and bone. When the plate is affixed to the bone, high shear force may appear between plate and bone, particularly near the end screws, which can lead to motion under weight-bearing. Two mechanisms might be involved in the prevention of motion: the bending stiffness of the screws or the friction between the screw and the plate. Experiments performed in vivo and in vitro show that with conventional plates, motion is prevented by friction and depends upon the axial force of the screw, pressing the plate onto the bone. The torque applied to the screws is crucial. Motion appears under smooth plates under relatively low physiological loads. With newly developed internal fixators, the motion is prevented by the structural stiffness of the plate-screw system. PMID- 11052378 TI - Mechanical analysis of the bone to plate interface of the LC-DCP and of the PC FIX on human femora. AB - The scope of this analysis was to evaluate the mechanical behaviour of newly developed plates at the junction between plate and bone (friction between plate and bone) for the limited contact dynamic compression plate (LC-DCP) and the point contact fixator (PC-Fix) under simulated physiological load and using the tension band principle on the human femur. The intact human cadaveric femora were plated on the lateral aspect according to the tension band principle (AO) and subjected to a load which simulated careful physiological load in single stance. Five strain gauges were glued around the bones, parallel to the bone axis, at five levels, whereby three of them had to be covered by a bone plate and the two others were just outside the plate location. The cross-sectional geometry had been obtained at these levels using computed tomography. One side was plated using the conventional compression plate LC-DCP and the contralateral side using the internal fixator PC-Fix. The LC-DCP was affixed using screws tightened at different torque values and the PC-Fix at a standard torque value. Motion (slippage) between the plate and the bone was indicated by a hysteresis of the strain reading during loading and unloading. Slippage was more important for the LC-DCP than for the PC-Fix, particularly at the proximal end of the plate and when the screws were insufficiently tightened on the LC-DCP. As expected, better stability was obtained with the PC-Fix. PMID- 11052380 TI - The point of view of the clinician: a prospective study of the mechanism of accidents and the morphology of tibial and fibular shaft fractures. AB - In a prospective study of 210 tibial shaft fractures, the accident mechanisms and the resultant fracture morphologies were analyzed. 86 fractures occurred due to indirect impact. The fracture morphology in this group consisted of short and long spiral fractures resulting from rotational injuries complicated by anterior torsion butterfly fragments if the person fell forward, posterior torsion butterfly fragments if they fell backwards and complicated by multiple torsion butterfly fragments if it was a high velocity injury. 124 fractures occurred due to direct impact. The fracture morphology in this group consisted of transverse, oblique segmental or crush fractures, complicated by one or more butterfly fragments due to bending, the injury depending on whether it was pure, one-point, three-point, or four-point-bending and on additional axial loading and velocity. In the indirect impact group, there were a few soft tissue injuries and fibular fractures at a different level to the tibial fracture. In the direct group, a large number of soft tissue injuries and fibular fractures at the level of impact were found. PMID- 11052379 TI - Strain distribution in plated and unplated sheep tibia an in vivo experiment. AB - After plate osteosynthesis changes in bone biology and bone mechanics are observed in the plated bone segment. Compromise of the vascular supply in the plate bed leads to a remodelling process and to a temporary porosity in the bone cortex underneath the plate. In addition, the plate takes over some of the physiological loading of the bone, which in turn alters the normal strain distribution of the cortical bone tissue. The aim of the present study is to determine the tissue deformation of the sheep tibia in vivo and the changes in tissue strain due to plating with plates of different rigidities. Measurements were performed on the intact bone at the mid diaphysis using the strain gauge technique. With different connections on the tension bridge (Wheatstone bridge), the strain was measured separately for pure axial loading, bending, and torsion before and after plating with a 4.5 mm stainless steel or titanium DCP. Under physiological load the sheep tibia is mainly deformed in torsion (62%) and bending (33%), and much less in axial loading (5%). Plating with a steel plate reduces the overall tissue strain by 18%, with a titanium plate by 13%. This reduction is mainly due to a reduction in axial tissue strain due to axial loading and bending and less to the reduction of tissue strain under torsion. In our in vivo model, plating with steel or titanium plates leads to a reduction of the physiological tissue strains. The difference between the different plates is small due to the fact that the high tissue strain under torsion is only slightly affected by plating. Thus, from the purely mechanical point of view and with regard to preserving normal tissue strains as much as possible, titanium plates offer little advantage compared with stainless steel plates. But, titanium as an implant material may offer advantages with respect to tissue compatibility and infection resistance. PMID- 11052381 TI - The point of view of the veterinary surgeon: bone and fracture. AB - It is important to understand the biomechanics of fractures if the morphological characteristics of fracture, fracture treatment and fracture healing are to be understood. Because of the mechanical properties of bone, which is stronger in compression than in tension, a fracture line is initiated perpendicular to the direction of tension or in shear. The mechanical properties of bone are also time dependent (viscoelasticity). Subsequently, when the load is applied at a higher load range, a greater amount of energy is stored within the bone, resulting in a more severe fracture (high energy fractures). There are five types of fracture: tension, compression, torque, bending, and shear, with typical morphologies: transverse fractures in tension, fracture line at 45 degrees to the long axis of the bone in compression, spiral fracture in torsion, combined transverse and butterfly fracture in bending, shear line when the overload is applied eccentrically. These different morphological types are shown on radiographs. PMID- 11052382 TI - The epidemiology of fractures of the proximal femur. AB - Between 1980 and 1989, reports on 21,145 fractures of the proximal part of the femur (15,428 women and 5,717 men) were collected by AO Documentation and are analysed in the present paper. The number of fractures increases exponentially with age for women and for men. Up to 50, the number of fractures is higher for men than for women and vice versa after 55 years. The exponential increase in the number of fractures with age appears for all the types A1 to B3 for both women and men. No special increase has been identified for women after 50 (expected age of menopause). PMID- 11052383 TI - The epidemiology of fractures of the distal femur. AB - Between 1980 and 1989, reports on 2,165 fractures of the distal part of the femur (1,051 women and 1,114 men) were collected by AO Documentation and are analyzed in the present paper. The number of fractures showed a bimodal pattern with a marked variation in the number of fractures in relation to gender and age. A larger prevalence of fractures was observed either in young men (about 20 years old, traffic or sport) and in old women (about 70, fall at home, osteoporosis). PMID- 11052384 TI - The epidemiology of diaphyseal fractures of the tibia. AB - AO Documentation collected reports of nearly five thousand diaphyseal tibial fractures occurring in the 1980s. The following conclusions can be drawn: i. The number of fractures affecting men is twice that affecting women. ii. These fractures occurred mainly in younger people (under 40 years of age) and no increase occurred in elderly people. iii. Most bending fractures occurred in young men (20-30 yrs) and the torsion fractures affected men and women of about 40 equally. iv. The number of simple fractures (type A) is about the same as for fractures with one fragment (type B). PMID- 11052385 TI - The mechanical strength of bones in torsion application to human tibiae. AB - How strong is a long bone in torsion? The principle of mechanical engineering of torsion on a beam is applied to the cylindrical tube model of a bone. The deformation and the strength of the tube are calculated. The calculated strength is about twice that of a real, not circular bone (human tibia) with the same cross-sectional area. PMID- 11052386 TI - A biomechanical enigma: why are tibial fractures not more frequent in the elderly? AB - Epidemiology revealed that diaphyseal fractures of the tibia affect young people, particularly young men; no increase was noticed for the elderly. This indicates that osteoporosis does not lead to increased bone fragility. Obviously, this is a biomechanical enigma. Torque measurements were carried out on human cadaveric tibiae and revealed a great correlation between the polar moment of inertia of the cortical bone at the tibial isthmus and the ultimate torque at failure (r = 0.83) and a lesser correlation between the cross-sectional density at the isthmus and the torque at failure (r = 0.57). Therefore, the size is more important than the degree of osteoporosis. We can speculate that endosteal resorption due to osteoporosis is compensated for by periosteal apposition and therefore does not lead to bone weakness. PMID- 11052387 TI - A sample of Chinese literature MRI diagnosis of interosseous membrane injury in Maisonneuve fractures of the fibula. AB - OBJECTIVE: In Maisonneuve fractures of the fibula (MFF) it remains controversial whether there is an injury of the interosseous membrane (IOM) and how severely it ruptures. The author studied injuries to the IOM in MFF by MRI to elucidate this question. METHOD: Twelve patients were examined with MRI before operation. MRI examination to the leg was performed with knee coil in 2 steps to obtain a complete image of the IOM. Using axial scan, we took T1 (TR500/TE40 msec), T1STIR and T2 sequence. The asymptote side was also examined in three patients as a comparison. RESULT: IOM was ruptured in a range of 32-112 mm, on average 79 mm proximal to the talar dome in all 12 patients. No IOM rupture was found at the level of high fibular fractures. IOM usually orients at the site 30 mm proximal to the talar dome. Anatomy and rupture of IOM are clearly visualized with T1 (TR500/TE40 msec) STIR sequence; haemorrhage and oedema would be seen better on a T2 image. CONCLUSION: The injury level of IOM in MFF is only at the distal 1/3 part of the leg; it is not consistent with high fibular fractures. MFF is a special type of pronation-external rotation type, there are four stages in the classification of MFF: 1) injury of medial structures, including medial malleolar fracture or rupture of the deltoid ligament; 2) rupture of the anterior tibiofibular ligament or avulsion fracture of one of its bone insertions, or one associated with interosseous ligament rupture and partial rupture of IOM in the distal 1/3 of the leg; 3) fracture of the proximal part of the fibula; 4) avulsion fracture of the posterior tibial tubercle. PMID- 11052388 TI - Evidence-based practice: the road best traveled? PMID- 11052389 TI - Well-being of parents of young children with asthma. AB - The relationships of family demands, caregiving demands, sense of coherence (SOC), and family hardiness (FH) with parents' well-being was evaluated in 76 families (75 mothers and 62 fathers) of young children (infant to 6 years) with asthma. The Resiliency Model of Family Stress, Adjustment and Adaptation (McCubbin, M., & McCubbin, 1993, 1996) was the conceptual framework for the study. The major hypothesis was that SOC and FH, separately and in combination, moderate both family system and caregiving demands on general well-being. With hierarchical regression analysis, SOC and FH explained 56% of the variance in mothers' well-being; family demands, SOC, and FH explained 67% of the variance in fathers' well-being. No moderating relationships were found for SOC or FH. Resiliency factors (SOC and FH) and family demands had direct relationships to the well-being of parents of young children with asthma. PMID- 11052390 TI - Narratives of family caregiving: four story types. AB - Researchers across disciplines have recognized considerable individual variation among caregivers in their response to the experiences of caregiving. One explanation for individual variation is that caregivers make different meanings from caregiving even under externally similar circumstances. This paper describes findings from a study that combined two qualitative strategies, across-case, thematic analysis and within-case, narrative analysis, to investigate meaning in accounts of family caregiving. Themes identified in the across-case analysis were interpreted in the context of patterns identified in the narrative analysis, as well as in the overall framework of caregivers' process of making meaning. Caregivers in this study told four types of stories: stories of ideal lives, stories of ordinary lives, stories of compromised lives, and ambiguous stories. Characteristics of each story type are described, and an example of an ambiguous story is also included as an illustration of the method. Findings suggest a new approach to understanding family caregiving that incorporates the diverse meanings caregivers make of their often similar experiences. PMID- 11052391 TI - Measuring Nursing Practice Models using Multi-Attribute Utility theory. AB - Nursing Practice Models (NPMs) represent the structural and contextual features that exist within any group practice of nursing. Currently, measurement of NPMs relies on costly and nonreproducible global judgments by experts. Quantitative measurement techniques are needed to provide a useful evaluation of nursing practice. Guided by Multi-Attribute Utility theory (MAU theory), an expert panel identified 24 factors representative of N PMs. The factors became elements in a computational index that, when summed, assigns a score to a given nursing unit reflecting the extent to which that unit's nursing practice model achieves the nursing professional ideal. Initial validation of the index and its elements consisted of comparing assessments of 40 nursing units generated by the index with a global evaluation provided by each of the expert panelists who proposed the model factors. Pearson correlations between the index-generated scores and the global assigned scores provided evidence supporting the preliminary validation of the index. PMID- 11052392 TI - Physical activity among older Mexican American women. AB - Theories of stage of readiness for change and self-efficacy guided this investigation of (a) performance of habitual physical activity (daily activity and leisure/sport activity), (b) preference for leisure Physical activities, and (c) relationships of age, stage of readiness for exercise, and exercise self efficacy with habitual physical activity. The sample consisted of 71 community residing Mexican American women aged 60-87 years. Fifty-six women reported performing at least one leisure/sport activity; the most frequent was walking, reported by 47 women. Fifty-three women stated that walking was their preferred leisure activity. Age, self-efficacy, and stage of readiness accounted for 27% of the variance in daily activity and 32% of the variance in leisure/sport activity. These findings suggest that stage of readiness for change and self-efficacy theories are useful for this population. PMID- 11052393 TI - An individualized intervention to overcome patient-related barriers to pain management in women with gynecologic cancers. AB - Concerns about reporting pain and using analgesics ultimately can contribute to poor pain management for many individuals. A nursing intervention to address these "patient-related barriers" was developed based on Johnson's self-regulation theory. The purpose of this pilot study was to determine whether provision of individually tailored sensory and coping information about analgesic side effects and specific information to counter misconceptions would enhance pain management in a sample of 43 women with gynecologic cancers. It was hypothesized that at 1 month post-test and 2-month follow-up, those subjects randomized to the information condition would (a) have lower barriers scores; (b) use more adequate analgesic medication; (c) have lower analgesic side effect scores; (d) have lower pain intensity scores; and (e) experience less pain interference with life and better overall quality of life compared to those in the care-as-usual control group. There was no main effect for group on any of the dependent variables. Rather, all women reported a decrease in barriers between baseline and 2-month follow-up (p<.05); all subjects experienced a decrease in pain interference with life scores between baseline and 1-month post-test (p<.05); and there was a significant shift of women from unacceptable pain management at baseline to acceptable pain management at 1-month post-test (p<.05). In addition, the majority of women reported that the intervention contained novel and useful information that helped them to feel more comfortable taking pain medication, to be less concerned about addiction, and helped them talk more openly about pain with a doctor or nurse. PMID- 11052394 TI - Psychometric properties of the Thai versions of State-Trait Anxiety Inventory for Children and Child Medical Fear Scale. AB - The purpose of this study was to investigate the psychometric properties of the Thai versions of the State-Trait Anxiety Inventory for Children (STAIC) and the Child Medical Fear Scale (CMFS) in Thai school age children. Subjects were 419 Thai students in grades 3-6 in three schools in Bangkok. Classrooms of students were randomly selected, and students within each classroom were divided into two groups to test the two instruments. Data were collected from students in the classrooms twice, 2 weeks apart. Content Validity Indices of the Thai STAIC and the CMFS were between 90-95%. Internal consistencies were > .80. Test-retest reliability coefficient for the Thai CMFS was .80, and coefficients for the STAIC were .62 for A-State scale and .68 for A-Trait scale. Both instruments seem to be promising anxiety/fear measures for Thai school age children; however, the Thai version of STAIC needs modification to enhance reliability and validity. PMID- 11052395 TI - Mediator and moderator variables in nursing research: conceptual and statistical differences. AB - Mediators and moderators are variables that affect the association between an independent variable and an outcome variable. Mediators provide additional information about how or why two variables are strongly associated. In contrast, moderators explain the circumstances that cause a weak or ambiguous association between two variables that were expected to have a strong relationship. Mediators and moderators are often overlooked in research designs, or the terms are used incorrectly. This article summarizes the conceptual differences between mediators and moderators. The statistical analysis of moderators and mediators in multiple regression is briefly described and two examples are presented. PMID- 11052396 TI - Transmission of viral respiratory infections in the home. AB - Respiratory viruses in the home exploit multiple modes of transmission. RSV is transmitted primarily by contact with ill children and contaminated objects in the environment. Influenza appears to be spread mainly by airborne droplet nuclei. Despite many years of study, from the plains of Salisbury, to the hills of Virginia, to the collegiate environment of Madison, WI, the precise routes rhinovirus takes to inflict the misery of the common cold on a susceptible population remain controversial. PMID- 11052397 TI - Transmission of rotavirus and other enteric pathogens in the home. AB - Rotavirus is the most common gastrointestinal pathogen present in day-care settings. Control and prevention of rotavirus infection are difficult because of the lack of a licensed vaccine, the absence of any effective treatment other than palliative measures and the presence of asymptomatic children shedding virus. Rotavirus is transmitted by fecal-oral contact and possibly by contaminated surfaces and hands and respiratory spread. Other gastrointestinal pathogens are also transmitted primarily by the fecal oral route, although contaminated surfaces, hands or food may also serve to transmit infection in some cases. Control and prevention measures for all enteric pathogens include isolating infected children from others, thoroughly cleaning and disinfecting environmental surfaces with effective agents and strictly following handwashing procedures before and after contact with infected persons and/or potentially contaminated surfaces. PMID- 11052398 TI - Transmission and control of infections in out-of-home child care. AB - The epidemiology of infections associated with out-of-home child care is well known, but our understanding of cost-effective prevention strategies is limited. Existing studies indicate that multidimensional interventional programs can reduce infection rates, but these conclusions are limited by a variety of methodologic concerns. Important areas for future research are to determine the critical elements of effective intervention programs, whether improvements in infection rates can be sustained over long periods and the costs of implementing and sustaining these programs. PMID- 11052399 TI - Relationship between cross-contamination and the transmission of foodborne pathogens in the home. AB - Environmental microbiologists have accumulated a substantial amount of information about the relationship between cross-contamination and the transmission of foodborne pathogens in the home. This information can be translated into guidelines for safe, effective hygienic practices in the home. Such guidelines cannot be mandated but should be strongly promoted via health and hygiene agencies to the general public. Establishing consensus-based, effective hygiene guidelines will benefit the community by reducing the risk of home-based infections such as foodborne disease and by lowering the associated healthcare costs. PMID- 11052400 TI - Consumer and market use of antibacterials at home. AB - In this increasingly complex, time-constrained world, consumers will continue to look for solutions that promise them peace of mind. A large component of this peace of mind is perceived as personal safety against infectious agents. Manufacturers have a responsibility to provide sound advice and to develop solutions to consumers' questions. Through working with leaders in the infection control field, as well as governmental organizations like the Centers for Disease Control and Prevention, the Food and Drug Administration and the Environmental Protection Agency, manufacturers can develop faster and more effective disinfectant and antimicrobial products. Targeted education programs are needed that clearly and effectively communicate proper infection control techniques and prudent use of antibacterial products to both the consumer and the health professional. Manufacturers should also work closely with the media to educate the public about the potential benefits and risks of their products. Finally manufacturers of household and personal cleaning products should help set guidelines for regulatory monitoring, including correct definition and use of common terms such as antibacterial, antimicrobial, antiviral and sanitization, as well as the extent of protection the consumer can expect from the product. PMID- 11052401 TI - Changing behavior to maintain a healthy home. AB - Multidimensional strategies involving interpersonal interaction, strategic alliances and targeted use of the media have been shown experimentally to be the most effective behavioral change programs. Health providers need to communicate with schools and community groups, as well as work with product manufacturers and consumer and health organizations. Together these groups can collaborate with the media to communicate accurate information and guidance to the public. Planning effective public health and behavioral change campaigns involves diagnosis and identification of the target behavior. The intervention message must be focused, tested and refocused. Effective communication, crucial to public health campaigns, uses multiple channels and provides frequent repetition of the message. PMID- 11052402 TI - Antibiotic and antiseptic resistance: impact on public health. AB - More and more we are moving patients from hospitals to homes for continued treatment. Vancomycin and triclosan were used for 30 years before any resistance emerged, because their applications were strictly limited. Today, after greatly increased use, resistance to both antibiotics and antibacterials has appeared. Of importance there are genetic links between resistance to antibiotics and to antibacterials. Health professionals and the public need to be educated about the rational use of drugs that affect the microbial world. The Alliance for the Prudent Use of Antibiotics, an international organization established in 1981 with members in more than 100 countries, has adopted education as its prime mission. Via its web site (www.apua.org) and linked information on reservoirs of antibiotic resistance (ROAR) among nonpathogenic bacteria, it reaches both providers and consumers. The message is simple: bacteria are needed for our survival. The vast majority of bacteria perform important functions that are crucial for our lives. Prudent use of both antibiotics and antibacterials must be championed to achieve and maintain the balanced microbial environment in which we have entered and evolved. PMID- 11052403 TI - Overview of Lysol scientific studies. AB - When followed, simple hygienic practices such as handwashing and surface disinfection along with proper food handling techniques can have a positive impact on the health of families and individuals in nonmedical settings like homes, day care and long term care facilities. Several studies have attempted to identify the specific role surface disinfectants can play in this effort. The evidence seems to indicate that these types of products, when properly used, can be beneficial. Because it has been established that environmental surfaces act as intermediates in the transmission of microorganisms throughout the day-care center and in homes, future studies should be developed to quantitate the impact of specific interventions in the reduction of microorganisms. PMID- 11052404 TI - Economic evaluation of infection control practices in day care and the home: methodologic challenges and proposed solutions. PMID- 11052405 TI - Diagnostic characteristics of 93 cases of a prepubertal and early adolescent bipolar disorder phenotype by gender, puberty and comorbid attention deficit hyperactivity disorder. AB - OBJECTIVE: Etiopathogenetic and treatment studies require homogeneous phenotypes. Therefore, effects of gender, puberty, and comorbid attention deficit hyperactivity disorder (ADHD) on DSM-IV mania criteria and other characteristics of a prepubertal and early adolescent bipolar disorder (PEA-BP) phenotype were investigated. METHOD: Consecutively ascertained PEA-BP (with or without comorbid ADHD) outpatients (n = 93) were blindly assessed by research nurses with comprehensive instruments given to mothers and children separately, consensus conferences, and offsite blind best estimates of both diagnoses and mania items. To fit the study phenotype, subjects needed to have current DSM-IV mania or hypomania with elated mood and/or grandiosity as one criterion and to be definite cases by severity ratings. RESULTS: Subjects were aged 10.9 +/- 2.6 years, had current episode length of 3.6 +/- 2.5 years, and had early age of onset at 7.3 +/ 3.5 years. No significant differences were found by gender, puberty, or comorbid ADHD on rates of mania criteria (e.g., elation, grandiosity, racing thoughts), mixed mania, psychosis, rapid cycling, suicidality, or comorbid oppositional defiant disorder (ODD), with few exceptions. Subjects with comorbid ADHD were more likely to be younger and male. Pubertal subjects had higher rates of hypersexuality. CONCLUSIONS: These findings support that the PEA-BP phenotype is homogeneous except for differences (hyperactivity, hypersexuality) that mirror normal development. PMID- 11052406 TI - Six-month stability and outcome of a prepubertal and early adolescent bipolar disorder phenotype. AB - OBJECTIVE: Six-month follow-up data are provided on a prepubertal and early adolescent bipolar disorder phenotype (PEA-BP). Stabilities were defined as continuous presence of PEA-BP and of individual mania criteria between baseline and 6 months. METHOD: Baseline and 6-month assessments of consecutively ascertained PEA-BP outpatients (n = 91) included comprehensive instruments given to mothers and children, separately, by research nurses; consensus conferences; and offsite blind best estimates of both diagnoses and mania items. To fit the study phenotype, subjects needed to have current DSM-IV mania or hypomania with elated mood and/or grandiosity as one mania criterion and to be definite cases by severity ratings. RESULTS: Of the 93 baseline subjects, 91 completed the 6-month assessment, for a retention rate of 97.8%. Baseline age was 10.9 +/- 2.7 years, and age of onset of current episode was 7.3 +/- 3.5 years. At 6 months, 85.7% still had full criteria and severity for mania or hypomania, and only 14.3% had recovered. Six-month stabilities of elated mood and grandiosity were high. Cox modeling and logistic regression did not show any significant effect of multiple covariates (e.g., gender, puberty, psychosis, mixed mania, rapid cycling, or naturalistic treatment). CONCLUSIONS: These longitudinal stability findings provide validation of a PEA-BP phenotype. Poor outcome was consistent with similarity of PEA-BP baseline characteristics to those of treatment-resistant adult-onset mania. PMID- 11052407 TI - Stimulant treatment in young boys with symptoms suggesting childhood mania: a report from a longitudinal study. AB - This study used data from a completed longitudinal study to examine the effects of methylphenidate on 6-12-year-old boys presumably at risk for bipolar disorder. Of 75 boys referred, diagnosed with hyperkinetic reaction of childhood (minimal brain dysfunction), treated clinically with methylphenidate, and followed as young adults, 23% (the maximorbid or MAX group) had childhood symptoms of irritability and emulated DSM-IV diagnoses of attention deficit hyperactivity disorder (ADHD), plus oppositional defiant or conduct disorder (ODD/CD) and anxiety or depression or both. The remaining boys (the minimorbid or MIN group) had fewer symptoms and disorders. MAX and MIN groups did not differ in rated response to methylphenidate, duration of treatment, clinically determined maintenance doses, concurrent or subsequent treatment with other medications, or other aspects of medication experience. At ages 21-23, individuals with bipolar related lifetime diagnoses (adult mania, hypomania, or cyclothymia) did not differ from those without bipolar-related diagnoses in any aspect of early methylphenidate treatment history. These findings indicate that ADHD boys with symptoms suggesting childhood mania do not respond differently to methylphenidate than boys without such symptoms, and there is no evidence here that methylphenidate precipitates young adult bipolar disorders in susceptible individuals. PMID- 11052408 TI - Therapeutic dilemmas in the pharmacotherapy of bipolar depression in the young. AB - Pediatric bipolar disorder is commonly mixed with co-occurring symptoms of major depression and mania. Knowledge has begun to accumulate on the treatment of the mania component, but limited information is available to guide the therapeutic approach to bipolar depression. To this end, we reviewed the medical charts of 59 patients with diagnosis of DSM-III-R bipolar disorder from an outpatient pediatric psychopharmacology clinic. Multivariate methods were used to model the probability of improvement and relapse at each visit of clinical follow-up. Serotonin-specific antidepressants were significantly associated with both an increased rate of improvement of bipolar depression-relative risk = 6.7 (1.9 23.6); p = 0.003-and a significantly greater probability of relapse of manic symptomatology-relative risk = 3.0 (1.2-7.8); p = 0.02. Although mood stabilizers improved manic symptomatology, they had no demonstrable effect on the symptoms of bipolar depression. Despite the increased risk of mood destabilization, serotonin specific antidepressants did not interfere with the antimanic effects of mood stabilizers. Because bipolar youth commonly come to clinical practice with depression, these results underscore the importance of assessing a lifetime history of bipolar disorder in making treatment decisions in depressed youth. PMID- 11052409 TI - A controlled study of nortriptyline in children and adolescents with attention deficit hyperactivity disorder. AB - OBJECTIVES: To study the efficacy and tolerability of nortriptyline (NT) in the treatment of pediatric attention deficit hyperactivity disorder (ADHD). METHODOLOGY: Subjects were outpatient children and adolescents with ADHD ascertained from clinical referrals. Subjects were enrolled in a 6-week open study in which NT was titrated to 2 mg/kg/day as tolerated over 2 weeks. Using either a 30 % reduction in the ADHD rating scale or a score of 1 or 2 on the Clinical Global Impression (CGI) scale for ADHD improvement, responders to treatment were then randomized into a 3-week, controlled discontinuation phase. During this phase, subjects either continued on their current dose of NT or were tapered to placebo under double-blind conditions. Subjects were monitored for symptoms of ADHD, oppositionality, anxiety, and depression. RESULTS: Of the 35 subjects enrolled in the study, 32 completed the open phase and 23 completed the discontinuation phase. The mean dose of NT was 80 mg (1.8 mg/kg/day), resulting in a serum level of 81 ng/ml. At the conclusion of the open 6-week study, NT was related to a significant reduction in ADHD (p < 0.001) and oppositional symptoms (p < 0.001). At the conclusion of the discontinuation phase, the 12 subjects randomized to NT had significantly lower scores on the DSM-IV ADHD symptom checklist than those 11 subjects randomized to placebo (31 versus 21; t = 2.2; p < 0.04). No significant adverse events were observed, and children were noted to have weight gain during the trial. CONCLUSIONS: These data suggest that NT is effective in reducing symptoms not only of ADHD but also of oppositionality. This group of children and adolescents tolerated robust dosing of NT well, with few clinical or cardiovascular adverse events. PMID- 11052410 TI - Pemoline treatment of adolescents with attention deficit hyperactivity disorder: a short-term controlled trial. AB - BACKGROUND: Despite the increased recognition of attention deficit hyperactivity disorder (ADHD) in adolescents, few controlled studies have assessed treatments for this age group. Adolescent issues, such as embarrassment at receiving medication at school and experimentation with abusable substances, have accelerated efforts to find effective, well-tolerated treatments beyond traditional stimulants. Pemoline has been found effective for treating both children and adults with ADHD but has not been evaluated in adolescents with ADHD. METHODS: Twenty-one adolescents (mean age 14 years old) diagnosed with ADHD by structured and clinical interviews participated in a 10-week, double-blind crossover design study of pemoline. Dosing was optimized with robust doses up to 3 mg/kg/day in one to two doses. Clinical evaluations of ADHD, depression, anxiety, and oppositional defiant disorder (ODD) symptoms were assessed weekly. RESULTS: Adolescents with ADHD exhibited a marked response to pemoline treatment relative to placebo on the ADHD rating scale (p = 0.001), with an average reduction of 3.02 points per week of treatment. Sixty percent of adolescents responded to pemoline, compared to 11% treated with placebo. This response was independent of gender or lifetime psychiatric comorbidity. Pemoline was well tolerated, with patients averaging 2.88 mg/kg/day in two doses per day, with a mean dose at end of follow-up of 181.1 mg (SD 45.6, range 112.5-262.5 mg). Side effects were mild, and no adverse hepatic events occurred. CONCLUSIONS: These findings resemble those reported in children and adults with ADHD. This trial suggests pemoline is well tolerated and effective in adolescents and may be a particularly useful ADHD treatment for adolescents. PMID- 11052411 TI - Adjunctive donepezil in attention deficit hyperactivity disorder youth: case series. AB - Despite well-documented improvement in attention deficit hyperactivity disorder (ADHD) in youths with the available single and combined pharmacologic agents, a number of youths remain with residual symptomatology causing impairment in multiple domains. Recent work has suggested a potential cognitive-enhancing role of cholinergic agents in ADHD. We describe five cases of ADHD youths aged 8-17 years being treated for ADHD with the acetylcholinesterase inhibitor donepezil (Aricept), all of whom demonstrated improvement. PMID- 11052412 TI - Sleep disturbances associated with attention deficit hyperactivity disorder: the impact of psychiatric comorbidity and pharmacotherapy. AB - OBJECTIVE: To estimate the relative risk of sleep problems associated with attention deficit hyperactivity disorder (ADHD), its pharmacotherapy, and its comorbid psychopathology. METHOD: Standard sleep questionnaires were used to assess sleep problems and characteristics in ADHD (n = 122) and non-ADHD (n = 105) comparison youths. RESULTS: ADHD was associated with 10 of 19 sleep problems assessed. However, after controlling for psychiatric comorbidity and pharmacotherapy with stimulants, the majority of these differences were no longer evident. Rather, treatment with stimulants and comorbidity with anxiety and behavior disorders were significantly associated with sleep disturbances. Positive family history of ADHD was associated with none of the sleep problems assessed here after adjusting for age, psychiatric comorbidity, and treatment with anti-ADHD medication. CONCLUSIONS: Although subjective sleep difficulties are common in ADHD youths, they are frequently accounted for by comorbidity and pharmacotherapy. Furthermore, the lack of an association between a positive family history of ADHD and sleep difficulties suggests that ADHD is not a misdiagnosis of the consequences of disruption of normal sleep. PMID- 11052413 TI - A discrete-time Lagrangian network for solving constrained quadratic programs. AB - A discrete-time recurrent neural network which is called the discrete-time Lagrangian network is proposed in this letter for solving convex quadratic programs. It is developed based on the classical Lagrange optimization method and solves quadratic programs without using any penalty parameter. The condition for the neural network to globally converge to the optimal solution of the quadratic program is given. Simulation results are presented to illustrate its performance. PMID- 11052414 TI - Tuning diversity in bagged ensembles. AB - In this paper, we investigate how the level of diversity amongst individual neural networks in a bagged ensemble can significantly influence overall ensemble generalization performance. We propose a new technique that tunes this diversity so that ensemble generalization performance is optimized and evaluate its performance on benchmark regression data-sets. PMID- 11052415 TI - On the capacity of multilayer neural networks trained with backpropagation. AB - The capacity of a layered neural network for learning hetero-associations is studied numerically as a function of the number M of hidden neurons. We find that there is a sharp change in the learning ability of the network as the number of hetero-associations increases. This fact allows us to define a maximum capacity C for a given architecture. It is found that C grows logarithmically with M. PMID- 11052416 TI - Neural fuzzy preference integration using neural preference Moore machines. AB - This paper describes preference classes and preference Moore machines as a basis for integrating different hybrid neural representations. Preference classes are shown to provide a basic link between neural preferences and fuzzy representations at the preference class level. Preference Moore machines provide a link between recurrent neural networks and symbolic transducers at the preference Moore machine level. We demonstrate how the concepts of preference classes and preference Moore machines can be used to interpret neural network representations and to integrate knowledge from hybrid neural representations. One main contribution of this paper is the introduction and analysis of neural preference Moore machines and their link to a fuzzy interpretation. Furthermore, we illustrate the interpretation and combination of various neural preference Moore machines with additional real-world examples. PMID- 11052417 TI - Cooperative coevolution of neural representations. AB - A genetic algorithm (GA) is used to search for a set of local feature detectors or hidden units. These are in turn employed as a representation of the input data for neural learning in the upper layer of a multilayer perceptron (MLP) which performs an image classification task. Three different methods of encoding hidden unit weights in the chromosome of the GA are presented, including one which coevolves all the feature detectors in a single chromosome, and two which promote the cooperation of feature detectors by encoding them in their own individual chromosomes. The fitness function measures the MLP classification accuracy together with the confidence of the networks. PMID- 11052418 TI - Instabilities and oscillation in the deterministic Boltzmann machine. AB - Simulations indicate that the deterministic Boltzmann machine, unlike the stochastic Boltzmann machine from which it is derived, exhibits unstable behavior during contrastive Hebbian learning of nonlinear problems, including oscillation in the learning algorithm and extreme sensitivity to small weight perturbations. Although careful choice of the initial weight magnitudes, the learning rate, and the annealing schedule will produce convergence in most cases, the stability of the resulting solution depends on the parameters in a complex and generally indiscernible way. We show that this unstable behavior is the result of over parameterization (excessive freedom in the weights), which leads to continuous rather than isolated optimal weight solution sets. This allows the weights to drift without correction by the learning algorithm until the free energy landscape changes in such a way that the settling procedure employed finds a different minimum of the free energy function than it did previously and a gross output error occurs. Because all the weight sets in a continuous optimal solution set produce exactly the same network outputs, we define reliability, a measure of the robustness of the network, as a new performance criterion. PMID- 11052419 TI - The epidemiology of respiratory tract infections. AB - Respiratory tract infections (RTIs) are the most common, and potentially most severe, of infections treated by health care practitioners. Lower RTIs along with influenza, are the most common cause of death by infection in the United States. Risk factors for pneumonia and other respiratory tract infections include: extremes of age (very young and elderly), smoking, alcoholism, immunosuppression, and comorbid conditions. The microbial cause of RTIs vary depending on the infection (i.e., pneumonia compared with acute bacterial sinusitis), setting (i.e., community-acquired compared with nosocomial), and other factors. The causative pathogens associated with CAP have changed in prevalence over time. Although Streptococcus pneumoniae remains the most common causative pathogen, a number of newer pathogens, such as Chlamydia pneumoniae and sin nombre virus, have been recognized in recent years. The emerging antimicrobial resistance of respiratory pathogens (most notably S. pneumoniae) has also increased the challenge for appropriate management of RTI. An awareness of the epidemiology and cause of specific respiratory infections should optimize care. PMID- 11052420 TI - Antimicrobial susceptibility patterns of respiratory pathogens--a global perspective. AB - Antimicrobial resistance among respiratory tract pathogens poses a major challenge for the ongoing use of antimicrobial agents for treating infected patients. Global antimicrobial susceptibility data has documented the existence of widespread resistance issues. Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis constitute the principal community-acquired respiratory tract bacterial pathogens. For H. influenzae, resistance to ampicillin varies from less than 5% in some European countries to greater than 30% in North America and Southeast Asia. For H. influenzae, resistance to trimethoprim/sulfamethoxazole has been shown to range from less than 5% in North America and Europe to greater than 25% in Europe, the Middle East, and India. For M. catarrhalis, 85% to 100% of isolates worldwide are beta-lactamase positive and, therefore, ampicillin and amoxicillin resistant. Penicillin-resistant S. pneumoniae shows considerable variability worldwide ranging from 6% to 80% whereas macrolide resistance among the pneumococci range from 0% to 90%. Clearly, documenting and understanding the emergence, dissemination, and infection with pathogens resistant to antimicrobial agents is essential for developing strategies to deal with this global problem. This article highlights the frequency of antimicrobial resistance among the respiratory pathogens from a global perspective. Also, mechanisms of resistance and factors associated with the emergence, dissemination, and colonization of resistant organisms are discussed. PMID- 11052421 TI - Evidence of bacterial infection in acute exacerbations of chronic bronchitis. AB - The frequency with which bacterial infection causes exacerbations of chronic obstructive pulmonary disease (COPD) may depend on the dominant pathology present; patients with chronic bronchitis are more susceptible to bacterial bronchial infections than those at the emphysema or asthma ends of the spectrum. However, impairment in respiratory function may be very important in governing the outcome of an exacerbation. Placebo-controlled trials have provided conflicting evidence of the efficacy of antibiotics in acute exacerbations. Overall, there is a significant benefit, particularly in certain patient groups, defined by symptoms and past history. Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis are the species most commonly isolated during exacerbations, and the same species may colonize the bronchial mucosa when the patient is in a stable state. Evidence is accumulating that bacteria are an independent stimulus of mucus hypersecretion and bronchial inflammation, and that they interact with other stimuli such as viral infection, atmospheric pollution, and tobacco smoke. New approaches are being used to investigate the importance of bacterial infection in patients with COPD. There are several good reasons why new more potent antibiotics might be expected to be superior to older standard compounds in the management of patients with problematic COPD. However, future studies should aim to confirm that bacteriologic superiority translates into improved clinical outcomes, and seek to measure the level of benefit. PMID- 11052422 TI - Treatment of acute and chronic rhinosinusitis. AB - Rhinosinusitis is a common health complaint that is often seen by primary care physicians and otolaryngologists in the United States. The complicated anatomy of the paranasal sinuses, as well as the multiple etiologies, contributes to the complexity that one often faces in trying to ameliorate or eradicate this disease in affected individuals. A full understanding of the fundamentals of rhinosinusitis, as well as the treatment options available for the different types, is important. It is very important for the physician to take an organized, step-by-step approach to the management of each patient with this complicated disease. As most cases of rhinosinusitis presenting to the generalist's office will be of viral origin, antibiotics should not be given unless the patient has purulent rhinorrhea or worsening symptoms lasting more than 5 days, or total symptoms lasting longer than 10 days. When medical treatment fails or is incomplete, adjunctive surgical treatment becomes an option. Generally, the symptoms that are most helped by surgery include persistent headaches, nasal obstruction, and recurrent or persistent purulent rhinorrhea unresponsive to medical management. Appropriate and timely referral for specialty care will result in the definitive management of recalcitrant rhinosinusitis when medical management alone fails or in cases where a complication or malignancy is suspected. This article reviews the current understanding of the anatomy, pathophysiology, classification, diagnosis, and potential complications of rhinosinusitis. It also describes the current approach to the treatment of both acute and chronic rhinosinusitis. PMID- 11052423 TI - Empiric therapy of community-acquired pneumonia. AB - Community-acquired pneumonia (CAP) remains a common and serious clinical problem despite the availability of potent antibiotics and aggressive supportive measures. The management of CAP begins with recognition of the likely pathogens causing the illness in the individual patient; identification of several simple clinical clues allows the organization of the broad variety of possible organisms into a more manageable list of likely pathogens. Once the most likely pathogens are identified, then initial antibiotics may be chosen to cover those possibilities. A number of treatment guidelines have been introduced in the past 10 years, most of which share more similarities than differences. We review 2 of the more established guidelines, those published by the American Thoracic Society (ATS) and the Infectious Diseases Society of America (IDSA). We also review the likely modifications of the ATS guidelines and discuss the impact of bacterial resistance on antibiotic choices. PMID- 11052424 TI - Antibiotic therapy in acute exacerbations of chronic bronchitis. AB - Chronic obstructive pulmonary disease (COPD) comprises a spectrum of conditions including chronic bronchitis, emphysema, asthma, and bronchiectasis. It has a prevalence in the United States of 5.1% to 5.4% in the middle-aged to elderly population, with a lower rate in nonsmoking individuals. Moreover, COPD is complicated by frequent and recurring acute exacerbations of chronic bronchitis (AECB). Overall, COPD represents the fourth leading cause of mortality in the United States and is the second leading cause of work disability. This condition is also associated with high morbidity and health care expenditures. Despite the controversy over the need to prescribe antibiotics for patients with AECB, high risk patients have been identified who will benefit from this therapy.These include, patients with a history of repeated infections (>4 per year), comorbid illnesses (such as diabetes, asthma, coronary heart disease), or marked airway obstruction. Furthermore, a bacterial cause is shown in approximately 50% of AECB episodes, and primarily includes Haemophilus influenzae, Moraxella catarrhalis, and Streptococcus pneumoniae. Additionally, resistance among community-acquired respiratory pathogens in the United States has risen dramatically, with beta lactamase production evident in 40% of H. influenzae and greater than 95% of M. catarrhalis isolates, and with approximately 10% of pneumococci highly resistant to penicillin and simultaneously resistant to macrolide antibiotics. The criteria used to make choices for antibiotic use in patients with AECB should include knowledge of the frequencies of pathogen resistance and patients' clinical characteristics. An effective antibiotic, however, must be able to rapidly resolve the acute infection with the least patient morbidity and need for hospitalization. Although there remains controversy as to when to initiate antibiotic therapy in patients with AECB, several guidelines have been published. PMID- 11052425 TI - Treatment of hospital-acquired penumonia. AB - Hospital-acquired pneumonia (HAP) remains a significant cause of morbidity and attributable mortality, especially among patients undergoing mechanical ventilation. The clinical approach to this disorder continues to evolve. Although our understanding of the epidemiology, risk factors, and pathogenesis of this disorder are expanding, consensus on diagnostic, therapeutic, and preventive strategies is lacking. Although outcome is significantly improved by the rapid introduction of appropriate antimicrobial therapy, presently available diagnostic tests rarely are able to identify a specific pathogen when antimicrobial choices are made. Thus, most therapy is by necessity empirical. The American Thoracic Society (ATS) published guidelines for the empiric treatment of HAP in 1996, this article reviews the recommendations of these guidelines and, if new information is available, updates these recommendations. PMID- 11052426 TI - An unusual case of upper-lobe pneumonitis. PMID- 11052427 TI - Heat preservation during cardiac surgery. PMID- 11052428 TI - A prospective comparison of three heat preservation methods for patients undergoing hypothermic cardiopulmonary bypass. AB - OBJECTIVE: To prospectively compare 3 methods of body heat preservation in patients undergoing surgery requiring the use of hypothermic cardiopulmonary bypass (CPB). DESIGN: Prospective, randomized, and nonblinded. SETTING: University teaching hospital. PARTICIPANTS: Adult cardiac surgery patients (n = 101). INTERVENTIONS: Subjects were randomly assigned to 1 of 3 treatment groups: Group 1 (n = 33) used a fluid-filled warming blanket, group 2 (n = 31) used a heated and humidified breathing circuit, and group 3 (n = 37) used intravenous fluid warmers for the administration of all fluids. Treatments started on separation from CPB and concluded at the end of the intraoperative experience. Anesthetic technique, minute ventilation, conduct of CPB, and room temperature in the operating room were standardized. MEASUREMENTS AND MAIN RESULTS: Blood temperature was measured at its nadir on CPB, on separation from CPB, and just before departure from the operating room. No differences were found among groups for CPB duration, coldest venous temperature on CPB, rewarming time, rate of rewarming, room temperature, or blood temperature on separation from CPB. There were no significant differences found in post-CPB temperature afterdrop among groups. CONCLUSIONS: This study suggests that there is no statistically significant disparity in the effectiveness of these 3 intraoperative heat preservation methods. Ease of use and cost-effectiveness should guide the choice of warming method post-CPB. PMID- 11052429 TI - Postcardiopulmonary bypass hypoxemia: a prospective study on incidence, risk factors, and clinical significance. AB - OBJECTIVE: To evaluate the clinical significance of low arterial oxygen tension inspired oxygen concentration (PaO2-FIO2) ratio, as a measure of hypoxemia, in the early period after cardiac surgery with cardiopulmonary bypass (CPB); and to evaluate the preoperative, intraoperative, and postoperative factors contributing to the development of hypoxemia within the first 24 hours after cardiac surgery with CPB. DESIGN: Prospective observational study. SETTING: University hospital. PARTICIPANTS: Patients who underwent elective or emergency cardiac surgery with CPB (n = 466). INTERVENTIONS: Preoperative clinical and laboratory data were recorded, as were intraoperative and postoperative data regarding the PaO2-FIO2 ratio, fluid and drug therapy, and chest radiograph. Data analysis evaluated hypoxemia as depicted by the PaO2-FIO2 ratios at 1, 6, and 12 hours after surgery. Thereafter, the effect of the PaO2-FIO2 ratios on time to extubation, lung injury, and length of hospital stay was evaluated. The risk factors were analyzed in 3 separate periods: preoperative, intraoperative, and postoperative. Univariate and multivariate analyses were performed on each period separately. All data were analyzed in 2 consecutive steps: univariate analysis and multivariate analysis. MEASUREMENTS AND MAIN RESULTS: PaO2-FIO2 ratios after CPB were significantly lower compared with baseline values. Six patients (1.32%) met the clinical criteria compatible with acute lung injury. All 6 patients had prompt recovery. Significant risk factors for hypoxemia were age, obesity, reduced cardiac function, previous myocardial infarction, emergency surgery, baseline chest radiograph with alveolar edema, high creatinine level, prolonged CPB time, decreased baseline PaO2-FIO2, use of dopamine after discontinuation of CPB, coronary artery bypass grafting, use of left internal mammary artery, higher pump flow requirement during CPB, increased level of hemoglobin or total protein content, persistent hypothermia 2 and 6 hours after surgery, requirement for reexploration, event requiring reintubation, and chest radiograph with alveolar edema 1 hour after surgery. Six hours after surgery, a lower PaO2-FIO2 ratio correlated significantly with time to extubation and lung injury. CONCLUSIONS: This study shows that despite improvements in the technique of CPB, hypoxemia depicted by low PaO2-FIO2 ratios is common in patients after CPB. It is short lived, however, and has minimal effect on the postoperative clinical course of these patients. PMID- 11052430 TI - Protective ventilation attenuates postoperative pulmonary dysfunction in patients undergoing cardiopulmonary bypass. AB - OBJECTIVE: To ascertain if protective ventilation can attenuate the damaging postoperative pulmonary effects of cardiopulmonary bypass (increases in airway pressure, decreases in lung compliance, and increases in shunt). DESIGN: Prospective, randomized clinical trial. SETTING: Single university hospital. PARTICIPANTS: Twenty-five patients undergoing elective coronary artery bypass graft procedure and early extubation. INTERVENTIONS: Thirteen patients received conventional mechanical ventilation (CV; respiratory rate, 8 breaths/min; tidal volume, 12 mL/kg; fraction of inspired oxygen [FIO2], 1.0; positive end expiratory pressure [PEEP], +5), and 12 patients received protective mechanical ventilation (PV; respiratory rate, 16 breaths/min; tidal volume, 6 mL/kg; FIO2, 1.0; PEEP, +5). Perioperative anesthetic and surgical management were standardized. Various pulmonary parameters were determined twice perioperatively: 10 minutes after intubation and 60 minutes after arrival in the intensive care unit. MEASUREMENTS AND MAIN RESULTS: The mean postoperative increase in peak airway pressure in group CV was significantly larger than the mean postoperative increase in peak airway pressure in group PV (7.1 v 2.4 cm H2O; p < 0.001). Group CV experienced significant postoperative increases in plateau airway pressure (p = 0.007), but group PV did not (p = 0.644). The mean postoperative decrease in dynamic lung compliance in group CV was significantly larger than the mean postoperative decrease in dynamic lung compliance in group PV (14.9 v 5.5 mL/cm H2O; p = 0.002). Group CV experienced significant postoperative decreases in static lung compliance (p = 0.014), but group PV did not (p = 0.645). Group CV experienced significant postoperative increases in shunt (15.5% to 21.4%; p = 0.021), but group PV did not (18.4% to 21.2%; p = 0.265). CONCLUSIONS: Data indicate that protective ventilation decreases pulmonary damage caused by mechanical ventilation in normal and abnormal lungs. The results of this investigation indicate that protective ventilation may also help attenuate the postoperative pulmonary dysfunction (increases in airway pressure, decreases in lung compliance, and increases in shunt) commonly seen in patients after exposure to cardiopulmonary bypass. PMID- 11052431 TI - High oxygen concentration exacerbates cardiopulmonary bypass-induced lung injury. AB - OBJECTIVE: To investigate the effect of ventilation with 100% oxygen on lung injury associated with surgery involving cardiopulmonary bypass (CPB). DESIGN: A prospective randomized study. SETTING: University hospital. PARTICIPANTS: Thirty patients undergoing coronary artery bypass graft surgery with CPB. INTERVENTIONS: Patients were randomized to receive 100% oxygen (Oxygen group) or 50% oxygen (Air group) throughout surgery. During CPB, patients' lungs in the Air group were flushed with air and in the Oxygen group with 100% oxygen. MEASUREMENTS AND MAIN RESULTS: Lung injury was evaluated by arterial oxygen tension-inspired oxygen concentration (PaO2-FIO2) ratio and cytokine levels (tumor necrosis factor-alpha and interleukin-8) in blood and bronchoalveolar lavage fluid measured before and after CPB. The lowest PaO2-FIO2 value was observed after 40 minutes following the completion of CPB in both groups. PaO2-FIO2 values 6 hours after CPB were not different from baseline in the Air group but remained lower (359+/-63 mmHg and 298+/-78 mmHg; p = 0.013) in the Oxygen group. Blood cytokine levels rose during surgery in both groups. Bronchoalveolar lavage levels of interleukin-8 did not change, whereas tumor necrosis factor-alpha increased only in the Oxygen group (p = 0.035). CONCLUSIONS: A significant decrease of oxygenation was observed in the early post-CPB period in both groups of patients, with delay in recovery in patients treated with 100% oxygen. A larger increase of the proinflammatory cytokines was found in patients treated with 100% oxygen. High oxygen concentrations during surgery with CPB should be used only when specifically required. PMID- 11052433 TI - Lidocaine for prevention of reperfusion ventricular fibrillation after release of aortic cross-clamping. AB - OBJECTIVE: To determine the efficacy of a bolus of lidocaine administered by way of the pump before releasing the aortic cross-clamp (ACC) in preventing the occurrence of reperfusion ventricular fibrillation. DESIGN: Prospective, randomized study. SETTING: University hospital. PARTICIPANTS: Patients undergoing coronary artery bypass graft surgery (n = 34). INTERVENTIONS: Seventeen patients received 100 mg of lidocaine by way of the pump 2 minutes before releasing the ACC, and a control group of 17 patients received 5 mL of normal saline. MEASUREMENTS AND MAIN RESULTS: In the control group, the incidence of reperfusion ventricular fibrillation was 70%, which was significantly decreased to 11% in the lidocaine group. A higher cardiac output after weaning from cardiopulmonary bypass was observed in the lidocaine group; this may be attributed to the lower incidence of reperfusion ventricular fibrillation and consequently the lower need for defibrillation by electric countershocks. CONCLUSIONS: The results suggest that a bolus of 100 mg of lidocaine administered 2 minutes before release of the ACC can safely decrease the incidence of reperfusion ventricular fibrillation and is associated with better hemodynamics after weaning from cardiopulmonary bypass. PMID- 11052432 TI - Postoperative atrial tachyarrhythmias in patients undergoing coronary artery bypass graft surgery without cardiopulmonary bypass: a role for intraoperative magnesium supplementation. AB - OBJECTIVE: To determine if intraoperative magnesium supplementation would be associated with a reduction in postoperative atrial tachyarrhythmias (POAT) in patients undergoing coronary artery bypass grafting (CABG) surgery without cardiopulmonary bypass (off-pump CABG surgery). DESIGN: Retrospective study. SETTING: University Medical Center. PARTICIPANTS: Patients who had undergone off pump CABG surgery (n = 124). INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The charts of 124 patients who had undergone off-pump CABG surgery (64 by anterior thoracotomy and 60 by median sternotomy) were retrospectively reviewed. Demographic data and perioperative care were recorded and compared among patients who did and did not experience POAT and among patients who did and did not receive intraoperative magnesium supplementation. Logistic regression analysis was used to assess the association between magnesium supplementation and incidence of POAT, controlling for other covariables. Of the 124 patients, 16 had a prior history of atrial or ventricular arrhythmias and/or were receiving antiarrhythmic medications. Medical records of the remaining 108 patients were reviewed. Twenty-four patients (22%) had POAT. Forty-two patients (39%) received intraoperative magnesium. In patients receiving intraoperative magnesium, the incidence of POAT was significantly decreased (12% v 29%; p = 0.03). In these patients, initial postoperative serum magnesium was significantly higher (2.37 mEq/L v 1.86 mEq/L; p < 0.01). In patients not receiving intraoperative magnesium, 35% had hypomagnesemia (serum magnesium < 1.8 mEq/L) compared with 9% of patients receiving magnesium (p < 0.01). Patients who received intraoperative magnesium and beta-adrenergic blockers had a lower incidence of POAT (5%) than patients who received only one (19%) or neither (33%) (p < 0.05). CONCLUSIONS: Intraoperative magnesium supplementation is associated with a decrease in POAT after off-pump CABG surgery. The combination of a beta-blocker and magnesium may reduce POAT further. It is recommended that intraoperative magnesium supplementation be part of the care of patients undergoing off-pump CABG surgery. PMID- 11052434 TI - Complete myocardial revascularization on the beating heart with epicardial stabilization: anesthetic considerations. AB - OBJECTIVE: To describe an anesthetic management protocol for patients undergoing cardiac surgery with multiple coronary artery bypass grafts without cardiopulmonary bypass (off-pump CABG surgery) by median sternotomy with mechanical stabilization. DESIGN: Retrospective nonrandomized analysis. SETTING: Tertiary care hospital. PARTICIPANTS: Sixty-six consecutive patients on whom off pump CABG surgery by median sternotomy was attempted. INTERVENTIONS: Anesthesia was induced with a combination of etomidate and fentanyl; pancuronium bromide was given for muscle relaxation; and anesthesia was maintained with isoflurane, desflurane, or sevoflurane in 100% oxygen. Maintenance of normothermia was attempted by keeping the room temperature at 70 degrees F, warming all fluids to 41 degrees C, and using 2.5 L/min of fresh gas flows and a heat and humidity exchanger. When available, a convective forced-air blanket was used to cover patients' head and shoulders. Patients who were not slated for revascularization of the circumflex vessels and who had good ventricular function received central venous pressure monitoring (26%); all other patients received a pulmonary artery catheter. MEASUREMENTS AND MAIN RESULTS: Of the 66 patients, 36% required an epinephrine infusion at a mean rate of 1.45+/-2.05 microg/min intraoperatively to maintain hemodynamic stability; 25% required inotropic support for < 12 hours in the intensive care unit. CONCLUSION: Institution of systematic hemodynamic management was associated with the successful completion of the surgical procedure in 61 patients (92%). Only 5 patients required conversion to regular CABG surgery with cardiopulmonary bypass. PMID- 11052435 TI - Temporal synthesis and release of endothelin within the systemic and myocardial circulation during and after cardiopulmonary bypass: relation to postoperative recovery. AB - OBJECTIVE: To determine endothelin levels in arterial, pulmonary, and myocardial vascular compartments in patients undergoing coronary artery bypass graft surgery and to examine the influence of endothelin on postoperative recovery. DESIGN: Prospective, clinical study. SETTING: University hospital. PARTICIPANTS: Fifty patients undergoing elective coronary artery bypass graft surgery. INTERVENTIONS: Endothelin plasma content (fmol/mL) was measured in 50 patients undergoing coronary revascularization from various vascular compartments before surgery and at specific intervals up to 24 hours postoperatively. MEASUREMENTS AND MAIN RESULTS: Myocardial endothelin gradient (coronary sinus - aorta) was calculated before cardiopulmonary bypass (CPB), at release of the aortic cross-clamp, immediately after CPB, and 0.5 hour after CPB. The requirement for inotropic therapy and duration of patient stay in the intensive care unit were determined. Systemic and pulmonary endothelin levels were increased by >80% immediately after CPB when compared with preoperative values and increased again by approximately 60% during the first 24 hours postoperatively (p < 0.05). The myocardial endothelin gradient was reversed after CPB, indicating myocardial production of endothelin (pre-CPB, -0.72+/-0.39 fmol/mL v 0.5 hour post-CPB, 0.60+/-0.49 fmol/mL; p < 0.05). Longer intensive care unit times (>28 hours) were associated with higher systemic endothelin levels when compared with shorter times (<18 hours) (16.30+/-1.33 fmol/mL v 9.81+/-1.67 fmol/mL; p < 0.05). Patients with higher endothelin levels 6 hours postoperatively had greater inotropic requirements during the intensive care unit period. CONCLUSION: Endothelin levels after CPB remained persistently increased for at least 24 hours after surgery and were associated with increased myocardial production of endothelin. These results suggest that the increased endothelin observed in the early postoperative period may contribute to a complex recovery from coronary artery bypass graft surgery. PMID- 11052437 TI - Parameters associated with perioperative baffle fenestration closure in the Fontan operation. AB - OBJECTIVE: To identify clinical parameters indicating perioperative fenestration closure in children who underwent the fenestrated Fontan operation. DESIGN: Retrospective. SETTING: Single children's hospital. PARTICIPANTS: Patients who underwent a fenestrated Fontan operation in 1996 through 1997 (n = 101). INTERVENTION: A fenestrated Fontan operation was performed in children with single-ventricle physiology. MEASUREMENTS AND MAIN RESULTS: Early perioperative closure of the fenestration occurred in 14 patients (group 1), whereas the fenestration remained patent in 87 patients (group 2). The groups were compared by the following parameters: demographics, cardiac catheterization and ultrasound data, and use of aspirin or warfarin preoperatively and intraoperatively by assessing the composition of the cardiopulmonary bypass solution, use of ultrafiltration and antifibrinolytics, protamine dose, last hematocrit on cardiopulmonary bypass, and requirement of blood products. Immediately postoperatively in the intensive care unit (ICU), cardiac filling pressures (central venous and left atrial pressure), coagulation profile, cardiac rhythm, chest tube drainage, length of stay in the ICU, and use of atrial pacing were reviewed. Significant indicators of early fenestration closure in this study as determined by multivariate stepwise logistic regression were a high transpulmonary pressure gradient (p = 0.015) and a higher oxygen saturation (p = 0.001) 1 hour after arrival in the ICU, a low fibrinogen level (p < 0.0001), and the need for temporary atrial pacing (p = 0.029). The fenestration was reopened in 13 patients in group 1. In 101 patients, there was no early mortality, and all patients survived to discharge. CONCLUSION: Factors that correlated with postoperative fenestration closure in the fenestrated Fontan operation in this study were a high transpulmonary pressure gradient and a high oxygen saturation 1 hour after arrival in the ICU, a low fibrinogen level, and the need for temporary atrial pacing. PMID- 11052436 TI - Assessment of sex hormone-binding globulin and osteocalcin in patients undergoing coronary artery bypass graft surgery. AB - OBJECTIVE: To determine sex hormone-binding globulin (SHBG) and osteocalcin (OC) levels in patients undergoing coronary artery bypass graft surgery to clarify the status of peripheral thyroid metabolism and to correlate SHBG and OC with thyroid hormones and adverse postoperative events. DESIGN: Prospective study. SETTING: University medical center. PARTICIPANTS: Fifty randomly selected patients undergoing coronary artery bypass graft surgery. INTERVENTIONS: On the morning of surgery before induction of anesthesia; 30 minutes after cross-clamping of the aorta; 2 hours and 6 hours after aortic cross-clamp removal; and on the first, second, third, and seventh postoperative mornings, blood samples were drawn and analyzed for OC, SHBG, triiodothyronine (tT3), free T3 (fT3), thyroxine (tT4), free T4 (fT4), thyroid-stimulating hormone, and thyroid-binding globulin. Adverse postoperative events were recorded. MEASUREMENTS AND MAIN RESULTS: Mean tT3 and fT3 decreased on average by 35% and 18% but remained within the normal range perioperatively. Similarly, mean SHBG and OC remained within the normal range. More than half of the patients investigated (60%) had OC concentrations below the normal range. Patients with pathologically decreased tT3 (n = 6) and tT4 (n = 16) intraoperatively and postoperatively had SHBG and OC concentrations similar to those in patients with normal tT3 and tT4 levels. Patients with postoperative complications had significantly lower OC levels preoperatively and on the first postoperative morning than those with an uneventful postoperative recovery. CONCLUSION: Despite significant intraoperative and postoperative decreases in levels of thyroid hormones, low T3 syndrome was rare in this patient population. Unchanged concentrations of SHBG and OC in patients with pathologically decreased tT3 or tT4 suggest normal local thyroid exposure at the tissue sites in these patients. OC may act as a predictor for postoperative outcome. PMID- 11052438 TI - Ketamine infusion versus isoflurane for the maintenance of anesthesia in the prebypass period in children with tetralogy of Fallot. AB - OBJECTIVE: To evaluate the use of ketamine in comparison with isoflurane in the maintenance of anesthesia in children with tetralogy of Fallot. DESIGN: Prospective, randomized study. SETTING: University hospital. PARTICIPANTS: Fifty children scheduled for correction of tetralogy of Fallot. INTERVENTIONS: After intubation, patients were assigned randomly to receive 2 different anesthesia maintenance regimens: group I, isoflurane, 0 to 1% plus fentanyl, 0.1 microg/kg/min; group II, ketamine, 0 to 5 mg/kg/h, plus fentanyl, 0.1 microg/kg/min. Isoflurane concentration and ketamine infusion rate were adjusted to maintain arterial pressure within 25% of baseline. Hemodynamic and respiratory parameters were recorded at the end of 4 intervals: T0, before induction of anesthesia; T1, induction to 10 minutes postintubation; T2, 10 minutes postintubation to poststernotomy; and T3, poststernotomy to completion of catheterizations. MEASUREMENTS AND MAIN RESULTS: In comparing group I with group II, significant differences were observed in mean arterial pressure (p < 0.0001), heart rate (p < 0.01), arterial oxygen saturation (p < 0.0001), arterial oxygen tension (p < 0.001), arterial carbon dioxide tension (p < 0.001), arterial pH (p < 0.0001), base excess (p < 0.05), and arterial to end-tidal carbon dioxide tension difference (p < 0.01) at T3. CONCLUSION: The use of ketamine anesthesia is recommended as an alternative maintenance regimen in children undergoing definitive correction of tetralogy of Fallot. PMID- 11052439 TI - Intraoperative recurrent laryngeal nerve monitoring during video-assisted throracoscopic surgery for patent ductus arteriosus. AB - OBJECTIVE: To develop a technique to identify and localize the recurrent laryngeal nerve (RLN) during video-assisted thoracoscopic surgery (VATS) for patent ductus arteriosus. DESIGN: Prospective clinical study. SETTING: Children's hospital. PARTICIPANTS: Sixty infants and children scheduled for elective closure of patent ductus arteriosus. INTERVENTIONS: With parental informed consent, 60 infants and children undergoing elective VATS for patent ductus arteriosus were studied. A thin, pencil-point, Teflon-coated, stimulating probe allowed direct stimulation (<2 mA, 100-msec pulse width) of the left RLN inside the thorax. A commercially available 4-channel neurologic monitor recorded compound evoked electromyograms (EMGs) from the left RLN and right RLN (as control) by needle electrodes placed percutaneously in the neck. Hoarseness, stridor, feeding difficulties, and voice changes were assessed postoperatively. MEASUREMENTS AND MAIN RESULTS: Left RLN EMGs were easily obtained in 59 of the 60 patients. The surgeon correctly identified the RLN visually once in the first 7 patients; this ability subsequently improved. EMG localization of the location or course of the RLN altered dissection, clip size, or clip position in 37 of 59 patients. CONCLUSION: Intraoperative EMG to identify location and route of the RLN was easy to perform, was effective in identifying RLN position, and appeared to facilitate dissection and clipping of the ductus. PMID- 11052440 TI - Echocardiographic assessment of coronary blood flow velocity during controlled hypotensive anesthesia with nitroglycerin. AB - OBJECTIVE: To determine the effect of nitroglycerin on coronary blood flow velocity during controlled hypotensive anesthesia in humans. DESIGN: Internally controlled prospective study. SETTING: Single university hospital. PARTICIPANTS: Twenty American Society of Anesthesiologists class I and II patients undergoing general anesthesia for surgical resection of a malignancy. INTERVENTIONS: General anesthesia was induced with thiopental, fentanyl, and succinylcholine and maintained with isoflurane and vecuronium. Transesophageal echocardiography was used to evaluate left ventricular wall motion and blood flow velocity in the left anterior descending coronary artery. Intravenous nitroglycerin was used to reduce systolic arterial pressure to 60 to 70 mmHg. Intravenous albumin 5% was administered to maintain pulmonary capillary wedge pressure >5 mmHg. MEASUREMENTS AND MAIN RESULTS: The left anterior descending coronary artery was visualized clearly in 16 of 20 patients. At a mean nitroglycerin dose of 16+/-14 microg/kg/min, peak diastolic left anterior descending flow velocity increased significantly from 32.5+/-10.3 cm/sec to 44.7+/-14.6 cm/sec (p = 0.0103). None of the patients developed any ST-segment changes. CONCLUSIONS: During nitroglycerin induced hypotensive anesthesia, coronary blood flow as assessed by peak diastolic left anterior descending flow velocity is preserved or increased in most patients. Increases in left anterior descending flow velocity are predictably achieved only at nitroglycerin doses >5 microg/kg/min. Intraoperative transesophageal echocardiography is useful in monitoring coronary flow velocity responses to controlled hypotensive anesthesia. PMID- 11052441 TI - Prophylactic nitroglycerin did not reduce myocardial ischemia during accelerated recovery management of coronary artery bypass graft surgery patients. AB - OBJECTIVE: To evaluate the use of a high dose of nitroglycerin (NTG) for prophylaxis against myocardial ischemia and infarction in patients undergoing coronary artery bypass graft (CABG) surgery with accelerated recovery. DESIGN: Prospective, double-blind, placebo-controlled randomized study. SETTING: A university-based medical center. PARTICIPANTS: Forty adult patients presenting for elective CABG surgery. INTERVENTIONS: Forty patients were divided into 2 blinded study groups. Twenty patients received 2 microg/kg/min of NTG starting before induction of anesthesia and continuing for 6 hours after extubation in the intensive care unit. The placebo group (n = 20) received normal saline during this same interval. MEASUREMENTS AND MAIN RESULTS: Hemodynamics, incidence and severity of myocardial ischemia, and myocardial infarction rates were determined. There were no differences in hemodynamic parameters between groups. The incidence of ischemia was approximately 35% in each group. Myocardial infarction (as determined by elevated creatine kinase-MB fraction, troponin I, and electrocardiogram criteria) was 10% in the placebo group and 5% in the NTG group (p = 0.234). CONCLUSIONS: This study shows a high incidence of myocardial ischemia and infarction in patients presenting for CABG surgery with an accelerated recovery management scheme. NTG was well tolerated clinically; however, it was not found to be protective against myocardial ischemia or infarction in this setting. PMID- 11052442 TI - Development of malignant hyperthermia during cardiopulmonary bypass. PMID- 11052443 TI - Use of a double-lumen endobronchial tube for severe bronchospasm during cardiopulmonary bypass. PMID- 11052444 TI - Resuscitation in pediatric balloon valvuloplasty: effects on cerebral perfusion and oxygenation. PMID- 11052445 TI - Management of a patient with atrioventricular septal defect and severe pulmonary hypertension undergoing major orthopedic surgery. PMID- 11052446 TI - Avulsion of an H graft during closed-chest cardiopulmonary resuscitation after minimally invasive coronary artery bypass graft surgery. PMID- 11052447 TI - Transesophageal echocardiographic evaluation of left ventricular function. PMID- 11052448 TI - Case 5--2000. Confusion of aortic valve and parts of an aortic root prosthesis during intraoperative transesophageal echocardiography. PMID- 11052449 TI - Pro: tracheal extubation should occur routinely in the operating room after cardiac surgery. PMID- 11052450 TI - Con: tracheal extubation should not occur routinely in the operating room after cardiac surgery. PMID- 11052451 TI - An unusual postoperative pulmonary complication. PMID- 11052452 TI - Interesting right ventricular transesophageal echocardiography findings. PMID- 11052453 TI - Off-pump coronary artery bypass graft surgery. PMID- 11052454 TI - Inaugural address: realizing the promise of health care equality: our leadership mandate for the new millennium. PMID- 11052455 TI - Clinical characteristics of community-dwelling black Alzheimer's disease patients. AB - There is a relative dearth of studies examining the cognitive and neuropsychiatric features of black Alzheimer's disease (AD) patients in the United States. Therefore, this cross-sectional investigation reported on the prevalence and clinical correlates of depression and psychosis in a community dwelling black AD sample. The study participants comprised 55 English-speaking black patients evaluated consecutively at a university-affiliated memory disorders clinic. All patients were evaluated utilizing standardized procedures and diagnosed with possible or probable AD according to the criteria established by the National Institute of Neurological and Communicative Diseases and Stroke Alzheimer's Disease and Related Disorders Association. The presence of neuropsychiatric symptoms, including major depression and psychosis (delusions or hallucinations) was established via a semistructured psychiatric interview with the patient and primary care giver. The level of global cognitive impairment was rated with the Mini-Mental State Examination. The results showed that major depression and psychosis were observed in 20% and 58% of the sample, respectively. Mood disturbance was linked with low education, whereas psychosis was associated with greater cognitive dysfunction. This study provides important insight into the clinical characteristics of community-dwelling black AD patients. It is clear that continued research in the area of ethnicity and dementia is warranted to better understand the clinical needs of blacks and other minority populations in the United States that are afflicted with AD. PMID- 11052456 TI - Automated oxyhemoglobin dissociation curve construction to assess sickle cell anemia therapy. AB - Oxyhemoglobin dissociation curves measure the most important function of red blood cells - the affinity for oxygen and its delivery to the tissues. This function may be deranged in sickle cell anemia and some other hemoglobinopathies. An automated oxyhemoglobin dissociation curve analyzer constructed dissociation curves in 55 patients with hemoglobinopathies and in 24 control subjects while maintaining constant temperature and pH. Sigmoid curves were converted to rectilinear ones using the Hill equation. Oxygen affinity of red cells was assessed by calculation of P50 (the PO2 at which hemoglobin is half saturated). Results revealed separation of oxyhemoglobin dissociation Hill plots according to phenotype but with wide variability. Mean oxygen affinity of fetal hemoglobin was greatest, whereas that of sickle hemoglobin was least. Other hemoglobins were intermediate. A positive correlation between decreased oxygen affinity and carboxyhemoglobin confirmed the decreased oxygen affinity of sickle hemoglobin and decreased oxygen affinity and increased diphosphoglycerate in red cells. Hill plots are less sensitive discriminators of oxygen affinity than traditional sigmoid dissociation curves and offer no particular advantage. Serial studies in a subset of three sickle cell anemia patients treated conservatively suggest automated oxyhemoglobin dissociation curves may be useful in assessment of effectiveness of newer therapies of sickle cell anemia after refinement of the method and studies of larger populations. PMID- 11052457 TI - Late middle-aged and older men living with HIV/AIDS: race differences in coping, social support, and psychological distress. AB - Although AIDS mental health research has recently devoted more attention to the psychosocial needs of older adults living with human immunodeficiency virus (HIV) disease, studies of this population have typically combined older African American and white participants into one large sample, thereby neglecting potential race differences. The current study examined race differences in stressor burden, ways of coping, social support, and psychological distress among late middle-aged and older men living with HIV/AIDS. Self-administered surveys were completed by 72 men living with HIV/AIDS in New York City and Milwaukee, WI (mean age = 53.4 years). Older African-American and white men experienced comparable levels of stress associated with AIDS-related discrimination, AIDS related bereavement, financial dilemmas, lack of information and support, relationship difficulties, and domestic problems. However, in responses to these stressors, older African-American men more frequently engaged in adaptive coping strategies, such as greater positive reappraisal and a stronger resolve that their future would be better. Compared to their African-American counterparts, HIV-infected older white men reported elevated levels of depression, anxiety, interpersonal hostility, and somatization. African-American men also received more support from family members and were less likely to disclose their HIV serostatus to close friends. As AIDS becomes more common among older adults, mental health-interventions will increasingly be needed for this group. The development of intervention programs for this group should pay close attention to race-related differences in sociodemographic, psychosocial, and behavioral characteristics. PMID- 11052458 TI - Prostate-specific antigen and androgens in African-American and white normal subjects and prostate cancer patients. AB - Prostate cancer in African Americans is more aggressive and common than in any other racial group. An endocrine mechanism has been proposed to account for this racial difference. However, androgen levels in African-American elderly normal subjects and prostate cancer patients have been insufficiently studied. Because the Albert Einstein Medical Center (AEMC) has a large African-American population, we could contribute racial data from which observations could be made within this study and in past and future studies. Blood from 38 screened men (mean age 65) with prostate-specific antigen (PSA) less than 4 ng/mL and normal rectal examination seen at the AEMC Cancer Center was studied using standard radioimmunoassays. The blood samples also served as our control. Our experimental group consisted of 51 prostate cancer patients (mean age 71 years), all of whom had nonmetastatic prostate cancer. Subjects were categorized by cancer status, race, and age group. In our screened subjects, PSA, testosterone, and dihydrotestosterone were not higher in African Americans than in whites. Furthermore, our prostate cancer patients demonstrated no significant racial variation for PSA, testosterone, and dihydrotestosterone. Our data also did not indicate any correlation between PSA and androgen levels in our cancer patients. In our population of elderly men, no racial differences in androgen levels were found. Androgen levels did not correlate with PSA levels in prostate cancer patients. PMID- 11052459 TI - Financial reimbursement: an incentive to increase the supply of transplantable organs. AB - The medical need for human organs suitable for transplantation far exceeds the current supply of organs and continues to grow. A perception exists that minorities, specifically African Americans, are not donating organs at a rate consistent with their representation of the national population. Cadaveric donor data from 1995 reveals that 11.4% of the donors were African American, 9.1% were Hispanic, and 1.5% were of Asian origin. The purpose of this study was to explore the hypothesis that financial reimbursement would increase organ donation among potential donors who are hesitant to donate for altruistic reasons alone. PMID- 11052460 TI - An unusual case of multiple cranial nerve palsies in Wegener's granulomatosis. AB - We describe an unusual case of Wegener's granulomatosis, which initially caused fulminant palsies affecting cranial nerves II, V, VI, VII, and VIII during a brief episode of the disease. The patient was successfully treated with immunosuppressive therapy. Wegener's granulomatosis should be suspected when multiple cranial nerves are initially affected. PMID- 11052461 TI - Granulomatous orchitis. PMID- 11052462 TI - The Utah paradigm of skeletal physiology: an overview of its insights for bone, cartilage and collagenous tissue organs. AB - In a 1960 paradigm of skeletal physiology, effector cells (chondroblasts, fibroblasts, osteoblasts, osteoclasts, etc.) regulated by nonmechanical agents wholly determined the architecture, strength, and health of bones, joints, fascia, ligaments, and tendons. Biomechanical and tissue-level phenomena had no roles in that paradigm. Subsequent studies and evidence slowly revealed skeletal tissue-level mechanisms and their functions, including biomechanical ones, as well as "game rules" that seem to govern them. That slow discovery process found that effector cells are only parts of tissue-level mechanisms, as kidney cells are only parts of nephrons and wheels are only parts of cars. Normally all those things help to determine skeletal architecture, strength, and health, and adding them to the 1960 paradigm led to the still-evolving Utah paradigm of skeletal physiology that concerns, in part, how load-bearing skeletal organs adapt to the voluntary mechanical loads on them. That caused controversies this article does not try to resolve; instead, it describes some issues they concern. In that regard, controversy can depend on how one assesses the relevance of facts to a problem more than on their accuracy. If a paradigm added new facts to a former one and the new one's advocates viewed all those facts as relevant, but the former's advocates questioned the relevance of some of the new facts, their views about a problem could differ even though each view depended on accurate facts. Readers would make their own judgments about the bearing of those ideas on this article's content. PMID- 11052463 TI - Effect of ethanol on bone mineral density of rats evaluated by dual-photon X-ray absorptiometry. AB - Abuse of alcohol may derange bone metabolism and cause osteoporosis. Due to confounding factors associated with alcohol abuse, e.g., dietary deficiencies and liver damage, a study using an animal model is preferable to examine whether alcohol itself actually reduces bone density. We evaluated the effect of alcohol intake on bone in rats by dual-energy X-ray absorptiometry. Six-week-old male (n = 16) and female (n = 16) Wister rats were divided into two groups. Sixteen alcohol-exposed rats (8 male and 8 female) were fed Lieber's liquid diet and 16 control rats (8 male and 8 female) were fed a control liquid diet. The bone mineral density (BMD) and bone mineral content (BMC) of the right femur were measured before and after experimental feeding under anesthesia. The BMD of lumbar spine (L2-L4) of sacrificed rats was measured. For male rats, BMD and BMC decreased significantly in the alcohol group (P = 0.0132 and 0.0133, respectively) but did not decrease in control group. For female rats, BMD and BMC decreased significantly in the alcohol group (P = 0.0012 and <0.0001, respectively) but did not decrease in the control group. For male rats, the mean ratio of BMD after experimental feeding divided by BMD before experimental feeding was significantly lower in the alcohol group than in the control group (P = 0.0031). For female rats, the mean ratio of BMD after experimental feeding divided by BMD before experimental feeding was also lower in the alcohol group than in the control group (P = 0.0002). For male rats, the mean BMD of L2-L4 after experimental feeding was significantly lower in the alcohol group than in the control group (P = 0.0210). For female rats, the mean BMD of L2-L4 after experimental feeding was also significantly lower in the alcohol group than in the control group (P = 0.0006). These results indicate that alcohol intake decreased the BMD of rats in both spongy and cortical bone, and that the reduction of BMD was greater in female rats than in male rats. PMID- 11052464 TI - Effects of phosphorus-containing calcium preparation (bone meal powder) and calcium carbonate on serum calcium and phosphorus in young and old healthy volunteers: a double-blinded crossover study. AB - To evaluate the effects of bone meal powder (BEC) on calcium and phosphorus metabolism, a calcium absorption test was conducted using a preparation of calcium carbonate (CAC) as the control drug. A total of 12 healthy volunteers, consisting of 6 younger (aged 20-29 years, 3 men and 3 women) and 6 older (aged 60-69 years, 3 men and 3 women) persons, were subjected to a double-blinded crossover study. Serum calcium (s-Ca) level significantly increased to 105.3% +/- 1.9% (P < 0.01 vs the basal value; mean +/- SD) from the basal value in the BEC group and to 104.4% +/- 2.7% (P < 0.01) in the CAC group at 3h post load. Urinary excretions of calcium (u-Ca/glomerular filtration rate, u-Ca/GF) after BEC and CAC load rose to 226.6% +/- 154.5% (P < 0.05) and 211.1% +/- 148.0% (P < 0.05), respectively. Serum phosphorus (s-P) levels after BEC load increased to 110.0% +/ 15.1% (P < 0.05), whereas that after CAC load showed no significant change (99.3% +/- 7.9%). On the other hand, urinary excretion of phosphorus (u-P/GF) after CAC load decreased to 60.0% +/- 32.4% (P < 0.01) and that in the BEC group showed no significant change (92.5% +/- 49.5%). The increase in s-Ca led to decrease in serum intact parathyroid hormone (i-PTH) level [77.3% +/- 33.4% (P < 0.05) for BEC and 69.5% +/- 20.3% (P < 0.01) for CAC] although s-P was increased by the BEC load. The responses to BEC and CAC administration were compared in the younger and the older groups. The responses in the younger and the older group showed fundamentally the same trends and to the same extent. However, the changes in serum ionized calcium (i-Ca) and i-PTH levels at 1.5 h post load were significantly smaller in the older group than in the younger group (P < 0.01; P < 0.05). The increment in s-P level after BEC load in the older group was larger than that in the younger group. In conclusion, BEC can modulate not only calcium but also phosphorus metabolism in both younger and older subjects. Further investigations are required to evaluate the effects of BEC on bone density and safety for renal function in long-term observations. PMID- 11052465 TI - Rheumatoid arthritis-related antigen 47kDa (RA-A47) is a product of colligin-2 and acts as a human HSP47. AB - We previously isolated RA-A47, which is recognized as an antigen of rheumatoid arthritis (RA), from a human chondrosarcoma-derived cell line (HCS-2/8). The N terminal 21-amino-acid sequence of RA-A47 had 81% homology to the deduced amino acid sequence of the human heat-shock protein (HSP) 47 gene, the colligin gene, and 100% homology to that of the colligin-2 gene. Moreover, as is HSP47, RA-A47 was a heat-inducible collagen-binding protein. To further characterize RA-A47, we isolated ra-a47 cDNA from HCS-2/8 cells and human periodontal ligament fibroblast (HPLF) cells. The isolated ra-a47 cDNAs from both cells were almost the same as that of colligin-2. C504 and G505 in the cDNA sequences of both cells and C598 in the cDNA of HCS-2/8 were different from the corresponding bases of colligin-2 cDNA. These differences were also observed in genomic DNA. colligin cDNA was not isolated. To show that the isolated cDNA encodes RA-A47 protein, it was expressed in Cos-7 cells. The produced protein was 47kDa and was recognized both with RA sera and antirat HSP47 antibody, indicating that it is RA-A47 and has structural similarity to HSP47. These results taken together with our previous findings show that RA-A47 is the putative colligin-2 gene product and behaves as a human HSP47. Although colligin has been considered the human HSP47 gene, failure to detect the colligin gene and its mRNA suggests that colligin does not exist in human cells and that the HSP47 gene is identical with colligin-2, which encodes RA-A47. PMID- 11052466 TI - Improvement of periarticular osteoporosis in postmenopausal women with rheumatoid arthritis by beta-alanyl-L-histidinato zinc: a pilot study. AB - The effect of zinc on bone metabolism in patients with rheumatoid arthritis (RA) is unknown. In the present pilot study, we investigated the effect of two antiulcer drugs, beta-alanyl-L-histidinato zinc (AHZ) and cimetidine, on bone metabolism in postmenopausal women with RA who had bilateral wrist pain. Eight patients were enrolled in a prospective, single-blind study consisting of 6-month cimetidine treatment (400 mg/day) followed by 6-month AHZ treatment (300 mg/day). Biochemical markers and bone mineral density (BMD) by dual energy X-ray absorptiometry were measured at baseline, 6 months, and 12 months. Three patients withdrew, and five patients (mean age 60: range 55-64 years) were analyzed. Their disease activity including wrist pain and dosages of prednisolone and disease modifying antirheumatic drugs remained unchanged during the 12-month treatment. The AHZ treatment increased serum zinc (AHZ vs cimetidine, +48.0% vs +5.6%), and resulted in significant increases of serum bone-specific alkaline phosphatase (+93.5% vs -14.7%) and BMD of the bilateral ultradistal radius (+4.9% vs -5.6%). However, the AHZ treatment had no effect on BMD of the lumbar spine (-2.0% vs +1.5%) or the bilateral distal third of radius (-2.1% vs +0.2%). In the AHZ treatment, the percentage change in BMD of the unilateral ultradistal radius with more severe wrist pain was positively correlated with the percentage change in serum zinc (r = 0.97). These findings suggest for the first time that AHZ treatment improves periarticular osteoporosis, probably through an increase of bone formation, in postmenopausal women with RA. Randomized double-blind controlled trials are needed. PMID- 11052467 TI - Effects of lower-leg lengthening on bone mineral density and soft tissue composition of legs in a patient with achondroplasia. PMID- 11052468 TI - Osteoclasts, integrins, and osteoporosis. PMID- 11052469 TI - Glucocorticoid-induced osteoporosis: pathogenesis and management. AB - Glucocorticoid- (GC-) induced osteoporosis and an increased risk of fractures are one of the most serious problems for patient using long-term GC therapy, such as those with rheumatoid arthritis, autoimmune diseases, inflammatory bowel diseases, bronchial asthma, and chronic lung diseases. GCs are known to affect both bone formation and resorption. In rheumatoid arthritis, the etiology of bone loss is multifactorial, including local inflammation around joints, release of bone-absorbing cytokines, physical inactivity, and malnutrition, in addition to the use of GC. Two guidelines have been published, by the American College of Rheumatology Task Force in 1966 and by the UK Consensus Group in 1998. Both guidelines recommend that patients 'receiving GC therapy at doses of 7.5 mg/day of prednisolone or more for 6 months or longer should have their bone mineral density measured and begin preventive therapies. Calcium and vitamin D supplements, sex hormone replacement, and weight-bearing exercise are the first line therapies. For patients who are unable to take sex hormone replacement therapy (HRT), bisphosphonates are recommended by both guidelines. In this article, we briefly summarize the pathogenesis of GC-induced osteoporosis and its prevention and treatment. PMID- 11052470 TI - Usefulness of magnetic resonance imaging for diagnosing deep anorectal abscesses. AB - PURPOSE: We evaluated the usefulness of magnetic resonance imaging for the preoperative diagnosis of deep anorectal abscesses. METHODS: Subjects were 21 patients with deep anorectal abscesses. Deep anorectal abscesses were classified into two types, ischiorectal and pelvirectal, according to their location. Patients were also classified into a single abscess group, which showed either an ischiorectal or pelvirectal abscess, and a double abscess group, which showed both ischiorectal and pelvirectal abscesses. The final diagnosis was made from surgical findings, and the types of deep anorectal abscesses determined by digital examination and magnetic resonance imaging were compared. RESULTS: Sensitivity of ischiorectal abscesses (20 lesions) with digital examination and magnetic resonance imaging was 75 and 95 percent, respectively, and that of pelvirectal abscesses (10 lesions) with digital examination and magnetic resonance imaging was 60 and 70 percent, respectively. Sensitivity of the magnetic resonance imaging was significantly higher than that of digital examination in ischiorectal abscesses. Diagnostic accuracy of digital examination and magnetic resonance imaging were both 83 percent in the single abscess group (12 patients), whereas in the double abscess group (9 patients) it was 22 and 78 percent, respectively. The rate of accurate diagnosis of magnetic resonance imaging compared with digital examination in the double abscess group was significantly higher than that in the single abscess group. CONCLUSION: Magnetic resonance imaging was useful for diagnosing and differentiating ischiorectal and pelvirectal abscesses. PMID- 11052471 TI - Evaluation of pelvic descent disorders by dynamic contrast roentgenography. AB - PURPOSE: For precise diagnosis and rational treatment of the increasing number of patients with descent of intrapelvic organ(s) and anatomic plane(s), dynamic contrast roentgenography of multiple intrapelvic organs and planes is described. METHODS: Sixty-six patients, consisting of 11 males, with a mean age (+/- standard deviation) of 65.6+/-14.2 years and with chief complaints of intrapelvic organ and perineal descent or defecation problems, were examined in this study. Dynamic contrast roentgenography was obtained by opacifying the ileum, urinary bladder, vagina, rectum, and the perineum. Films were taken at both squeeze and strain phases. On the films the lowest points of each organ and plane were plotted, and the distances from the standard line drawn at the upper surface of the sacrum were measured. The values were corrected to percentages according to the height of the sacrococcygeal bone of each patient. From these corrected values, organ or plane descents at strain and squeeze were diagnosed and graphically demonstrated as a descentgram in each patient. RESULTS: Among 17 cases with subjective symptoms of bladder descent, 9 cases (52.9 percent) showed roentgenographic descent. By the same token, among the cases with subjective feeling of descent of the vagina, uterus, peritoneum, perineum, rectum, and anus, roentgenographic descent was confirmed in 15 of 20 (75 percent), 7 of 9 (77.8 percent), 6 of 16 (37.5 percent), 33 of 33 (100 percent), 25 of 37 (67.6 percent), and 22 of 36 (61.6 percent), respectively. The descentgrams were divided into three patterns: anorectal descent type, female genital descent type, and total organ descent type. CONCLUSIONS: Dynamic contrast roentgenography and successive descentgraphy of multiple intrapelvic organs and planes are useful for objective diagnosis and rational treatment of patients with descent disorders of the intrapelvic organ(s) and plane(s). PMID- 11052472 TI - Relationship between microsatellite instability and telomere shortening in colorectal cancer. AB - PURPOSE: Two pathways have been proposed for the development of colorectal cancers: loss of heterozygosity and replication error. Colorectal cancers arising through the replication error pathway, like most hereditary nonpolyposis colorectal cancers, show microsatellite instability. It has been also reported that telomere shortening frequently occurs in colorectal cancers and that telomerase is often activated strongly in them. The aim of this study was to examine whether any relationships can be found among microsatellite instability, telomere length, and telomerase activity in colorectal cancers. METHODS: Genomic DNA was extracted from 55 invasive cancers and corresponding normal mucosas. Five microsatellite loci were analyzed by polymerase chain reaction. Telomere length was examined by Southern blot analysis. Telomerase activity was assayed by telomeric repeat amplification protocol with minor modifications. RESULTS: Microsatellite instability was found in 8 (14.5 percent) of 55 tumors, and all of them showed short telomeres. Furthermore, four high-frequency microsatellite instability tumors that showed microsatellite instability at more than two loci exhibited remarkably short telomeres. The microsatellite instability correlated significantly with frequency of telomere shortening (P = 0.0183; Fisher's exact probability test), but not with strength of telomerase activity. CONCLUSION: The relationship identified by this study between microsatellite instability and telomere shortening might suggest some association between the DNA mismatch repair system and the telomere maintenance mechanism in colorectal cancers. PMID- 11052473 TI - Natural history of early colorectal cancer: evolution of a growth curve. AB - PURPOSE: There are very few studies on the development of early colorectal cancers, although we have previously reported growth speeds of early colorectal cancer in a radiographic retrospective study. The aim of this study was to estimate a statistical curve for cancer growth from mucosal cancer. METHODS: Subjects of the study were 31 patients with cancer in which initial lesions were diagnosed as mucosal cancer. These lesions were overlooked in the first or second investigations, but were detected later. Initial radiographic features were as follows; 4 pedunculated lesions, 1 semipedunculated lesion, 6 sessile lesions, 9 superficially elevated lesions, and 11 superficially depressed lesions. The diameters of the initial lesions were 12.1+/-6.1 mm. The final depths of invasion were 6 mucosal cancers, 12 submucosal cancers, 6 muscularis propria cancers, and 7 serosal cancers. The observation period between the initial and final examination was 41.5+/-25.8 months. The growth curve was estimated by an exponential curve with the natural logarithm of d = e (a + b x t), where a is the intercept (initial tumor size) and b is the regression coefficient (growth speed). RESULTS: A growth curve was obtained as follows: diameter = 12.5 x 2(t/77) (r = 0.448, P < 0.0001) and 95 percent confidence interval of time = 53 to 173 months. Subsequently, volume = 1 x 10(3) x 2(t/26), and the 95 percent confidence interval of time = 18 to 58 months. CONCLUSION: Growth speed of early colorectal cancer was estimated through a statistically significant growth curve. Estimated doubling time of the volume of early colorectal cancer was 26 (95 percent confidence interval, 18-58) months. From these results we could obtain a rational cancer surveillance program using appropriate procedures with different sensitivities. PMID- 11052474 TI - Apoptosis and its regulation in flat-type early colorectal carcinoma: comparison with that in polypoid-type early colorectal carcinoma. AB - PURPOSE: The aim of this study was to investigate the relationship among apoptotic cell death, proliferative activity, and the expression of apoptosis regulating proteins (p53, p21 (WAF1/CIP1), and bax) in flat-type early colorectal carcinoma and to compare these factors with those in polypoid-type early colorectal carcinoma. METHODS: Formalin-fixed, paraffin-embedded tissues of 11 flat-type early colorectal carcinomas and 17 polypoid-type early carcinomas were studied. The histologic diagnosis was either well-differentiated adenocarcinoma or carcinoma in adenoma, and the depth of invasion was limited to mucosa or submucosa. Apoptotic cells were detected by terminal deoxynucleotide transferase mediated deoxyuridine triphosphate-biotin nick end-labeling method, and proliferative activity was determined by Ki-67 immunohistochemistry using monoclonal antibody MIB-1. Apoptosis-regulating proteins were determined by immunohistochemistry using antibody DO-7 (p53), Cip1 (p21 (WAF1/CIP1)), and Bax (bax). RESULTS: There was no significant difference in terminal deoxynucleotide transferase-mediated deoxy-uridine triphosphate-biotin nick end-labeling index between flat-type early colorectal carcinoma and polypoid-type early carcinoma, at 1.9 vs. 1.1, respectively. In flat-type carcinoma terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling index in the p53 protein overexpression group was significantly smaller than that in the p53 protein-negative group (P < 0.05). The Ki-67 labeling index/terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate-biotin nick end labeling index ratio in the p53 protein overexpression group was significantly higher than that in the p53 protein-negative group (P < 0.05). In polypoid-type carcinoma, the terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling index and Ki67/terminal deoxynucleotide transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling index ratio showed no significant difference between the p53 protein overexpression group and p53 protein-negative group. CONCLUSION: p53-dependent apoptosis may contribute to the development of flat-type early colorectal carcinoma. Apoptosis and its regulation in flat-type early colorectal carcinoma may differ from those in polypoid-type carcinoma. PMID- 11052475 TI - Analysis of the PTEN gene mutation in polyposis syndromes and sporadic gastrointestinal tumors in Japanese patients. AB - PURPOSE: PTEN is a candidate tumor suppressor gene for mutations which are responsible for Cowden disease. Recently, it has been shown that PTEN is mutated in several human neoplasms. To investigate the role of PTEN in tumorigenesis, we screened its mutation in Japanese patients with gastrointestinal polyposis and various sporadic tumors. METHODS: The entire coding region of PTEN was screened by single strand conformational polymorphism or direct sequencing for somatic mutations in 16 gingival papillomas, 4 juvenile polyps, 10 esophageal papillomas, and 20 colorectal cancers and for germline mutations in three patients with Cowden disease (including one with Lhermitte-Duclos disease) and one patient each with juvenile polyposis syndrome, Turcot's syndrome, and Cronkhite-Canada syndrome. RESULTS: Germline mutations were found in all cases of Cowden disease. One mutation was a nonsense mutation at codon 130 (CGA-->TGA), and the other two were splice site mutations at the 5' site of intron 4 and the 3' site of intron 8. We could not detect germline mutations in other patients with gastrointestinal polyposis or somatic mutations in sporadic tumors. CONCLUSIONS: We confirmed previous reports that germline mutations in PTEN are responsible for Cowden disease. However, somatic mutations of PTEN may not play a major role in tumorigenesis, at least in colorectal cancers, esophageal papillomas and gingival papillomas. PMID- 11052476 TI - High proliferative activity is associated with dysplasia in ulcerative colitis. AB - PURPOSE: Ulcerative colitis is associated with an increased risk of colorectal neoplasia. Markers of proliferation are reported to be valuable in the diagnosis of dysplasia in ulcerative colitis. However, it is not known whether dysplastic change or proliferative change occurs first. Whether abnormal proliferation is present in normal-seeming mucosa in ulcerative colitis was investigated. METHODS: Eighteen cancer or high-grade dysplasia specimens and 9 low-grade dysplasia specimens from 5 patients and 51 specimens from 31 patients without neoplasia were studied. Immunostaining with anti-Ki 67 antibody was used to evaluate proliferative activity. Labeling index (in the superficial one-half of crypt) was calculated. Crypts with labeling index more than 0.3 were determined to have abnormal proliferation. RESULTS: The mean +/- standard error of the mean labeling index in specimens negative for dysplasia (0.056+/-0.004) was significantly lower than that in low-grade dysplasia specimens (0.418+/-0.024) and that in high-grade dysplasia specimens (0.503+/-0.027; P < 0.0001). In specimens negative for dysplasia, only 4 (4 cases) of 339 (1.2 percent) crypts had abnormal proliferation, whereas the ratio of crypts with abnormal proliferation was 76 percent (54/71) in low-grade dysplasia and 92.1 percent (35/38) in high-grade dysplasia. The labeling index in background mucosa was 0.139+/-0.009, which was significantly higher than that in specimens negative for dysplasia (P < 0.001). In background mucosa 15.7 percent of crypts showed abnormal proliferation. A follow-up study revealed that two of four cases developed cancer or high-grade dysplasia one and seven years after proliferative abnormality was detected in nondysplastic specimens. CONCLUSION: Ki-67 immunostaining can be an aid in the diagnosis of dysplasia. High proliferating activity in background mucosa suggests that proliferating activity change precedes dysplasia detected with hematoxylin and-eosin staining. PMID- 11052477 TI - Histologic grade of metastatic lymph node and prognosis of rectal cancer. AB - PURPOSE: It is important to identify cases with a high risk of recurrence to improve the prognosis of colorectal cancer. In this study the difference between the histology of the primary lesion and that of the metastatic lymph node was investigated in an attempt to identify the cases with a high risk of recurrence. METHODS: One-hundred eighty-five patients with Dukes C rectal cancer who had undergone curative resection were investigated. The histologic grade of the metastatic lymph node was determined and compared with other clinicopathologic factors to determine its significance as a prognostic factor. RESULTS: The histologic grade was the same between the primary lesion and the metastatic lymph node in 46.2 percent of all cases, although in the group with well-differentiated adenocarcinoma at the primary lesion the concordance was only 29.5 percent. In the group with well-differentiated adenocarcinoma at the primary lesion, the five year survival rate was 75.3, 64, and 25 percent in the groups with well differentiated, moderately differentiated, and poorly differentiated adenocarcinoma at the metastatic lymph node, respectively. The differences between the survival rates of well-differentiated and poorly differentiated adenocarcinoma at the metastatic lymph node were statistically significant (P < 0.05). According to multivariate analysis the histologic grade of primary lesion was the most significant prognostic factor (hazard ratio: 2.2801, P = 0.0008). However, in well-differentiated adenocarcinoma of patients with Dukes C rectal cancer at the primary lesion, the histology of metastatic lymph node was also an important prognostic factor. CONCLUSIONS: It is clear that the histologic grade between the primary lesion and metastatic lymph node was frequently different, especially in the group with well-differentiated adenocarcinoma at the primary lesion. The analysis of the metastatic lymph node was considered to have additional importance for the prediction of prognosis. PMID- 11052478 TI - Predictive factors for detecting colorectal carcinomas in surveillance colonoscopy after colorectal cancer surgery. AB - PURPOSE: The aim of this study was to identify the high-risk groups for metachronous colorectal carcinoma among patients who undergo colorectal cancer surgery. METHODS: Three hundred forty-one patients undergoing colorectal cancer surgery who had undergone surveillance colonoscopy at least twice during a period of more than three years were analyzed. A metachronous colorectal carcinoma was defined as a new colorectal carcinoma detected by surveillance colonoscopy after surgery. RESULTS: Surveillance colonoscopy was performed 4.6 times per patient during an average of 6.2 years. Twenty-two metachronous colorectal carcinomas in 19 patients were detected, and 14 (64 percent) of 22 were detected within five years of surgery. The cumulative incidence of developing colorectal carcinomas during a five-year period was 5.3 percent. Seventeen (77 percent) of 22 carcinomas were 10 mm or less in size. Ten (71 percent) of the 14 carcinomas in early stages showed a flat appearance. Univariate analysis showed that extracolonic malignancy, coexistence of adenoma, and synchronous multiple colorectal carcinoma were significant predictive factors for detecting colorectal carcinomas in surveillance colonoscopy and that family history of colorectal carcinoma was a possible predictive factor. Multivariate analysis performed with Cox proportional hazards regression model showed that extracolonic malignancy and the coexistence of adenoma were significant predictive factors. CONCLUSION: We recommend that patients with the above predictive factors receive surveillance colonoscopy meticulously and regularly. PMID- 11052479 TI - Effect of early postoperative feeding on the healing of colonic anastomoses in the presence of intra-abdominal sepsis in rats. AB - PURPOSE: Intra-abdominal infection is generally considered a major risk factor for dehiscence of primary colon anastomosis. To elucidate the indications for nutritional support during intra-abdominal sepsis, we investigated the healing of anastomoses in an animal model. METHODS: Twenty male Sprague-Dawley rats (280-320 g) underwent cecal ligation and single puncture. After 24 hours the perforated cecum was removed, and the left colon was transected and anastomosed in a single layer inverted fashion. Animals were randomly assigned to receive both chow and water (early-fed group; n = 10) or water alone for the first 72 hours and chow thereafter (late-fed group; n = 10). Colon-bursting pressure was measured five days after the anastomosis, at which time the anastomosis was excised. RESULTS: The survival rate after cecal ligation and single puncture was 100 percent, and blood cultures were positive in 20 percent of animals five days after surgery. All data are expressed as means +/- standard error of the mean. Body weight increased more in the early-fed group than in the late-fed group (15.6+/-3 vs. 6.3+/-2.8 g; P < 0.001). Early feeding resulted in increased anastomotic bursting pressure (200+/-11 vs. 161+/-12 mmHg; P < 0.05) and total collagen concentration at the site of anastomosis (2.36+/-0.09 vs. 2.01+/-0.07 microg/mg wet tissue; P < 0.01) compared with the late-fed group. CONCLUSION: Early feeding has a positive effect on anastomotic healing in the presence of intraabdominal sepsis. The mechanism by which early feeding enhances the colonic anastomotic healing is unclear, although preservation of colonic collagen seems to play a significant role. PMID- 11052480 TI - Lateral node dissection and total mesorectal excision for rectal cancer. AB - PURPOSE: Although the existence of lateral lymphatic drainage of the rectum has been verified anatomically, the clinical importance of it has not yet been fully investigated. The lack of a definition of lateral lymphatic flow makes it difficult to analyze and compare data. The aim of this study was to define the concept of lateral lymphatic drainage and explore its relationship to total mesorectal excision and to disclose the incidence and efficacy of dissection of lateral node involvement. METHODS: Review of anatomic and clinical research on lateral lymphatic flow was made to create a definition of lateral lymphatic flow. Based on this review, a three-space dissection was designed and applied. A retrospective analysis was made of 764 patients with rectal cancer treated by a curative three-space dissection operation during 20 years starting in 1975 at Cancer Institute Hospital. RESULTS: Lateral lymphatic flow passes from the lower rectum and through the lateral ligament laterally beyond the mesorectum. It then ascends along the internal iliac artery and, in addition, inside the obturator space. Sixty-six cases proved to have lateral node involvement, which comprised 8.6 percent of all rectal cancer and 16.4 percent of low-lying (lower margin below 5 cm above the dentate line) rectal cancer cases. The five-year survival rate of these 66 cases was 42.4 percent. There were 16 cases that had a solo lateral node involvement. CONCLUSION: Lateral lymphatic flow from low-lying rectal cancer passes outside the boundaries of total mesorectal excision but within the range of curative surgery by three-space dissection. PMID- 11052481 TI - Sphincter-preserving techniques for anal fistulas in Japan. AB - After the articles by Eisenhammer, Parks, Hanley, and Goligher and the classification system established by Sumikoshi, surgeons in Japan became very active in creating and modifying new sphincter-preserving surgical procedures for low intersphincteric and ischiorectal fistulas. This article reviews the literature, summarizes each procedure, identifies the transitions, and discusses recurrence rates, cures, and subsequent variations. Japanese surgeons favor sphincter-preserving techniques for low intersphincteric fistulas that involve partial or complete coring out of fistula tracts. Common sphincter-preserving procedures for ischiorectal fistulas involve coring-out techniques and muscle filling procedures. The recurrence rates of sphincter-preserving procedures for low intersphincteric fistulas are between 5 and 6 percent, whereas the recurrence rates for the sphincter-preserving techniques used on ischiorectal fistulas are in the 7 to 10 percent range. PMID- 11052482 TI - Screening for colorectal cancer: current status in Japan. AB - PURPOSE: Screening for colorectal cancer using a guaiac-based fecal occult blood, or Hemoccult, test has been demonstrated to reduce colorectal cancer mortality. However, the magnitude of effectiveness is relatively low because of poor sensitivity of the Hemoccult test. The immunochemical fecal occult blood test has been shown to be much more sensitive than the Hemoccult test in detecting preclinical colorectal cancer in an asymptomatic population. The purpose of this article is to discuss the validity of the immunochemical fecal occult blood test and the efficacy of a population-based screening program using the test. METHODS: Relevant articles were primarily identified through MEDLINE search. Review was focused on the studies of population screening programs with the immunochemical fecal occult blood test. RESULTS: Sensitivities for colorectal cancer calculated in the same population were reported to be 67 to 89 percent and only 33 to 37 percent for the immunochemical test and Hemoccult test, respectively. Case control studies and other observational studies showed that screening programs using the immunochemical fecal occult blood test by hemagglutination reaction would reduce the risk of dying of colorectal cancer by 60 percent or more for those screened annually compared with those unscreened. It was also shown that a screening strategy using the immunochemical fecal occult blood test had the best cost-effectiveness ratio among the methods available. Nearly 5 million persons are currently screened per year in Japan, yielding 0.15 to 0.2 percent colorectal cancer cases among persons with positive fecal occult blood test results. CONCLUSIONS: These results strongly suggest that a screening program with immunochemical fecal occult blood test has promising advantages in terms of effectiveness over programs with the Hemoccult test. More stress is warranted on introduction of immunochemical fecal occult blood testing as a screening test in place of the guaiac fecal occult blood test. PMID- 11052483 TI - Crohn's disease in Japan: diagnostic criteria and epidemiology. AB - New diagnostic criteria for Crohn's disease and a review of Japanese epidemiologic studies are presented. New diagnostic criteria for Crohn's disease were established by the Research Committee of Inflammatory Bowel Disease, set up by the Japanese Ministry of Health and Welfare. For a definite diagnosis one of the following three conditions is required: 1) longitudinal ulcer or luminal deformity induced by longitudinal ulcer or cobblestone pattern, 2) intestinal small aphthous ulcerations arranged in a longitudinal fashion for at least three months plus noncaseating granulomas, and 3) multiple small aphthous ulcerations in both the upper and lower digestive tract not necessarily with longitudinal arrangement, for at least three months, plus noncaseating granulomas. Moreover, ulcerative colitis, ischemic enterocolitis, and acute infectious enterocolitis should be excluded. Data from the Japanese Ministry of Health and Welfare, in addition to data collected from two study groups, these being the two largest studies in Japan, are reviewed with regard to epidemiology. The number of patients with Crohn's disease has increased remarkably. The prevalence and the annual incidence of patients with Crohn's disease in Japan were estimated to be approximately 2.9 and 0.6 per 10(5) population in 1986, respectively, and 13.5 and 1.2 per 10(5) population in 1998. Characteristic features of Crohn's disease in Japan are that the male-female ratio exceeds 2, and that there is no second peak of incidence in the age group of 55 to 65 years. Clinically, Crohn's disease with only multiple small aphthous ulcerations, which is the earliest stage of the disease that is diagnosable, was found in 5 percent of patients. PMID- 11052484 TI - Laparoscopic ultralow anterior resection combined with per anum intersphincteric rectal dissection for lower rectal cancer. AB - Rectal resection with total mesorectal excision is perhaps the most technically challenging of laparoscopic procedures, and the purpose of this study was to show that laparoscopic ultralow anterior resection is feasible for lower rectal cancer. Seven patients with lower rectal cancer were treated in this way with a satisfactory outcome in each case, and on the basis of this limited study, we suggest that extension of laparoscopy to the treatment of very low rectal lesions may be of advantage. PMID- 11052485 TI - Invasive colon cancer derived from a small superficial depressed cancer: report of a case. AB - A 64-year-old male in May 1997 was diagnosed by colonoscopy and a barium enema examination as having an invasive cancer in the transverse colon. Pathologic study of the resected surgical specimen revealed a well-differentiated adenocarcinoma invading the muscularis propria. He had a colonoscopic examination in 1991 and was diagnosed as having multiple adenomas, which were endoscopically removed. After that he had annual colonoscopy or barium enema examination follow ups. At endoscopy in February 1994, a superficial depressed cancer 6 mm in diameter had been detected. However, the cancer was not seen again in several endoscopic examinations until one in 1997. Because the location of the lesion detected in 1994 and that of the invasive carcinoma detected in 1997 were identical, it was considered that the superficial depressed cancer developed, 40 months later, to an advanced cancer. Doubling time was calculated as 8.4 months. PMID- 11052486 TI - Decorin antisense gene therapy improves functional healing of early rabbit ligament scar with enhanced collagen fibrillogenesis in vivo. AB - Injured ligaments heal with scar tissue, which has mechanical properties inferior to those of normal ligament, potentially resulting in re-injury, joint instability, and subsequent degenerative arthritis. In ligament scars, normal large-diameter collagen fibrils have been shown to be replaced by a homogenous population of small collagen fibrils. Because collagen is a major tensile load bearing matrix element and because the proteoglycan decorin is known to inhibit collagen fibrillogenesis, we hypothesized that the restoration of larger collagen fibrils in a rabbit ligament scar, by down-regulating the proteoglycan decorin, would improve the mechanical properties of scar. In contrast to sense and injection-treated controls, in vivo treatment of injured ligament by antisense decorin oligodeoxynucleotides led to an increased development of larger collagen fibrils in early scar and a significant improvement in both scar failure strength (83-85% improvement at 6 weeks; p < 0.01) and scar creep elongation (33-48% less irrecoverable creep; p < 0.03) under loading. This is the first report that in vivo manipulation of collagen fibrillogenesis improves tissue function during repair processes with gene therapy. These findings not only suggest the potential use of this type of approach to improve the healing of various soft tissues (skin, ligament, tendon, and so on) but also support the use of such methods to better understand specific structure-function relationships in scars. PMID- 11052487 TI - Compressive compared with tensile loading of medial collateral ligament scar in vitro uniquely influences mRNA levels for aggrecan, collagen type II, and collagenase. AB - To test the hypothesis that loading conditions can be used to engineer early ligament scar behaviors, we used an in vitro system to examine the effect that cyclic hydrostatic compression and cyclic tension applied to 6-week rabbit medial collateral ligament scars had on mRNA levels for matrix molecules, collagenase, and the proto-oncogenes c-fos and c-jun. Our specific hypothesis was that tensile stress would promote more normal mRNA expression in ligament whereas compression would lead to higher levels of mRNA for cartilage-like molecules. Femur (injured medial collateral ligament)-tibia complexes were subjected to a hydrostatic pressure of 1 MPa or a tensile stress of 1 MPa of 0.5 Hz for 1 minute followed by 14 minutes of rest. On the basis of a preliminary optimization experiment, this 15-minute testing cycle was repeated for 4 hours. Semiquantitative reverse transcription-polymerase chain reaction analysis was performed for mechanically treated medial collateral ligament scars with use of rabbit specific primer sets for types I, II, and III collagen, decorin, biglycan, fibromodulin, versican, aggrecan, collagenase, c-fos, c-jun, and a housekeeping gene, glyceraldehyde-3 phosphate dehydrogenase. Cyclic hydrostatic compression resulted in a statistically significant increase in mRNA levels of type-II collagen (171% of nonloaded values) and aggrecan (313% of nonloaded values) but statistically significant decreases in collagenase mRNA levels (35% of nonloaded values). Cyclic tension also resulted in a statistically significant decrease in collagenase mRNA levels (66% of nonloaded values) and an increase in aggrecan mRNA levels (458% of nonloaded values) but no significant change in the mRNA levels for the other molecules. The results show that it is possible to alter mRNA levels for a subset of genes in scar tissue by supplying unique mechanical stimuli in vitro and thus that further investigation of scar engineering for potential reimplantation appears feasible. PMID- 11052488 TI - The pentapeptide NKISK affects collagen fibril interactions in a vertebrate tissue. AB - The pentapeptide NKISK has been reported to inhibit the binding of decorin, a proteoglycan on the surface of collagen fibrils, to fibronectin, a tissue adhesion molecule. Because of our interest in fibril-fibril binding as it relates to changes in length of ligament or tendon (during growth or contracture), we investigated the potential of this peptide to dissociate fibrils. The peptide permitted the release of intact fibrils into suspension for examination under the electron microscope (which has not previously been possible in mature vertebrate tissues). PMID- 11052489 TI - Tenascin-C in tendon regions subjected to compression. AB - Tendon regions subjected almost exclusively to tension differ from regions subjected to high levels of compression as well as tension. Regions not exposed to compression consist primarily of spindle-shaped fibroblasts surrounded by densely packed longitudinally oriented collagen fibrils formed principally from type-I collagen. In contrast, regions subjected to compression have a fibrocartilagenous structure and composition: they consist of spherical cells surrounded by a matrix containing hyaline cartilage proteoglycans (aggrecan) and type-II collagen as well as type-I collagen. Reducing their adhesion to the matrix may help cells in the latter regions establish and maintain a spherical shape and minimize their deformation when the tissue is subjected to mechanical stress. We hypothesized that expression of tenascin-C, an anti-adhesive protein, is part of the adaptation of tendon cells to compression that helps establish and maintain fibrocartilagenous regions. To test this hypothesis, we compared segments of bovine flexor tendons subjected to repetitive compression (distal) with segments that are not subjected to compression (proximal) to determine whether they differed in tenascin-C content and expression. RNA and protein analyses showed that tenascin-C expression was elevated in the distal tendon. Tendon cells from the distal segment expressed more tenascin-C mRNA than did cells from the proximal segments for as long as 4 days in cell culture, indicating that increased tenascin-C expression is a relatively stable feature of the distal cells. Moreover, purified tenascin-C inhibited the attachment of cultured tendon cells to fibronectin. These observations support the hypothesis that tenascin-C expression is a cellular adaptation to compression that helps establish and maintain fibrocartilagenous regions of tendons by decreasing cell matrix adhesion. PMID- 11052490 TI - Insulin-like growth factor-I is expressed by avian flexor tendon cells. AB - Cells in normal tendon are in a resting G0 state, performing maintenance functions. However, traumatic injury introduces growth factors such as platelet derived growth factor and insulin-like growth factor from blood as well as activates endogenous growth factors. These factors stimulate migration and proliferation of tendon cells at the wound area. Tendon cells require growth promoting factors to transit the cell cycle. To evaluate the contribution of endogenous growth factors in tendon, extracts of the epitenon and internal compartment of avian flexor tendon as well as medium of cultured cells from the epitenon (tendon surface cells) and internal tendon (tendon internal fibroblasts) were collected to assess their ability to stimulate DNA synthesis. Acid-ethanol extracts of tissues and medium were chromatographed on a P-30 molecular sieve column and assayed for mitogenic activity by quantitating [3H]thymidine incorporation into tendon cell DNA. The extract from the internal tendon compartment was more stimulatory for DNA synthesis than that from the epitenon, particularly when tested on tendon internal fibroblasts. However, conditioned medium fractions from surface epitenon cells stimulated DNA synthesis to a high degree on both tendon surface cells and tendon internal fibroblasts. Conditioned medium from tendon internal fibroblasts was also stimulatory. An anti-insulin like growth factor-I antibody ablated most of the mitogenic activity present in both tissues and conditioned medium. The levels of acid-extractable insulin-like growth factor-I in tendon were determined by competitive radioimmunoassay as 1.48+/-0.05 ng/g tissue for the epitenon and 3.83+/-0.03 ng/g tissue for the internal compartment. Results of Western immunoblots of conditioned medium revealed insulin-like growth factor-I at the 7.5 kDa position. Cultured tendon surface cells and tendon internal fibroblasts as well as cells in intact flexor tendon expressed insulin-like growth factor-I mRNA detected by reverse transcriptase-polymerase chain reaction. In situ hybridization histochemistry positively identified insulin-like growth factor-I mRNA in tendons from 52-day old chickens. Platelet-derived growth factor was not detected at the protein or message levels. Furthermore, tendon surface cells and tendon internal fibroblasts both expressed receptors for insulin-like growth factor-I detected by flow cytometry. These data suggest that tendon cells express insulin-like growth factor-I mRNA and synthesize insulin-like growth factor-I in both the epitenon and the internal compartment of tendon, which is present in an inactive form, most likely bound to insulin-like growth factor-binding proteins. PMID- 11052491 TI - Migration of cells from human anterior cruciate ligament explants into collagen glycosaminoglycan scaffolds. AB - Regeneration of the human anterior cruciate ligament after complete rupture offers several theoretical advantages over reconstruction, including maintenance of the complex insertion sites and fan-shape of the ligament and preservation of remaining proprioceptive fibers within the ligament substance. Well vascularized connective tissues, such as dermis, heal as a result of migration of fibroblasts into a provisional scaffold, the fibrin clot. Wound closure is subsequently facilitated by a contractile cell phenotype. This study was designed to determine if fibroblasts intrinsic to the human anterior cruciate ligament were capable of migrating from their native extracellular matrix onto an adjacent provisional scaffold in vitro. Another objective was to determine whether any of the cells that successfully migrated into the scaffold expressed the contractile actin isoform, alpha-smooth muscle actin, associated with wound contraction in other tissues. The results demonstrated that the cells intrinsic to the human anterior cruciate ligament were able to migrate into a collagen-glycosaminoglycan scaffold, bridging a gap between transected fascicles in vitro. As a result of this cell migration and proliferation, areas in the scaffold contained cell number densities similar to those seen in the human anterior cruciate ligament in vivo. No extracellular matrix or tissue formation was seen in the gap between directly apposed transected ends of the anterior cruciate ligament explants cultured without an interposed collagen-glycosaminoglycan scaffold. The fascicle collagen-glycosaminoglycan-fascicle constructs and the fascicle-fascicle explants displayed minimal adherence after 6 weeks in culture. Any disruption in the contact area between explant and scaffold, even as small a gap as 50 microm, prevented cell migration from the explant to the collagen-glycosaminoglycan scaffold at the area of loss of contact. All cells that migrated into the scaffold at early time periods expressed the alpha-smooth muscle actin isoform. These results demonstrate that cells that migrate into and proliferate within the collagen-glycosaminoglycan matrix have contractile potential as reflected in their expression of the alpha-smooth muscle actin isoform. The role of these contractile cells in the healing process warrants further investigation. Moreover, this study demonstrates the potential of cells intrinsic to the human anterior cruciate ligament to migrate into collagen-glycosaminoglycan scaffolds that may ultimately be investigated as implants to facilitate ligament healing and regeneration. PMID- 11052492 TI - Bilateral proprioceptive defects in patients with a unilateral anterior cruciate ligament reconstruction: a comparison between patients and healthy individuals. AB - Proprioception of the knee was measured in 20 patients with reconstructed anterior cruciate ligaments and in 19 age-matched controls. The mean time from surgery was 2 years. Three tests of proprioception were used: (a) threshold to detection of passive motion from 20 and 40 degrees toward flexion and extension, (b) active reproduction of a 30 degrees passive angle change, and (c) visual reproduction of a 30 degrees passive angle change. The aim was a complete, bilateral, proprioceptive evaluation of patients who had undergone reconstruction of the anterior cruciate ligament. As compared with those in the control group, the knees with reconstructed anterior cruciate ligaments had a higher threshold to detection of passive motion in the extension trials from 20 and 40 degrees (p = 0.0003 and 0.04, respectively) and in the flexion trials from 20 and 40 degrees (p = 0.004 and 0.0008, respectively). When the uninjured knees of the patients were compared with those in the control group, higher values for threshold to detection of passive motion were found in the flexion trials from 20 degrees (p = 0.002) and 40 degrees (p = 0.02). Thus, decreased proprioceptive ability was present in some measurements of these patients after reconstructive surgery, not only in injured knees but also in uninjured knees, as compared with the reference group. The functional relevance of these findings was not investigated in this study, but the results suggest that bilateral proprioceptive considerations should be made when evaluating prognostic factors, treatment, and risk of contralateral knee injury in patients with reconstructed anterior cruciate ligaments. PMID- 11052493 TI - Knee pain and joint loading in subjects with osteoarthritis of the knee. AB - Although treatments for osteoarthritis of the knee are often directed at relieving pain, pain may cause patients to alter how they perform activities to decrease the loads on the joints. The knee-adduction moment is a major determinant of the load distribution between the medial and lateral plateaus. Therefore, the interrelationship between pain and the external knee-adduction moment during walking may be especially important for understanding mechanical factors related to the progression of medial tibiofemoral osteoarthritis. Fifty three subjects with symptomatic radiographic evidence of osteoarthritis of the knee were studied. These subjects were a subset of those enrolled in a double blind study in which gait analysis and radiographic and clinical evaluations were performed after a 2-week washout of anti-inflammatory and analgesic treatment. The subjects then took a nonsteroidal anti-inflammatory drug, acetaminophen, or placebo for 2 weeks, and the gait and clinical evaluations were repeated. The change in the peak external adduction moment between the two evaluations was inversely correlated with the change in pain (R = 0.48, p < 0.001) and was significantly different between those whose pain increased (n = 7), decreased (n = 18), or remained unchanged (n = 28) (p = 0.009). Those with increased pain had a significant decrease in the peak external adduction (p = 0.005) and flexion moments (p = 0.023). In contrast, the subjects with decreased pain tended to have an increase in the peak external adduction moment (p = 0.095) and had a significant increase in the peak external extension moment (p = 0.017). The subjects whose pain was unchanged had no significant change in the peak external adduction (p = 0.757), flexion (p = 0.234), or extension (p = 0.465) moments. Thus, decreases in pain among patients with medial tibiofemoral osteoarthritis were related to increased loading of the degenerative portion of the joints. Additional long-term prospective studies are needed to determine whether increased loading during walking actually results in accelerated progression of the disease. PMID- 11052494 TI - Comparison and reproducibility of fast and conventional spoiled gradient-echo magnetic resonance sequences in the determination of knee cartilage volume. AB - Quantitation of articular cartilage by magnetic resonance imaging and three dimensional reconstruction has been validated and is likely to be a useful outcome measure in clinical trials of arthritis. The cost of magnetic resonance imaging is largely dependent on scanning time. The aim of this study was to compare a fast spoiled gradient-echo sequence magnetic resonance imaging scanning protocol, which takes 5 minutes and 44 seconds, with the standard, previously validated spoiled gradient-echo sequence protocol, which takes 11 minutes and 56 seconds, in the measurement of knee cartilage volume. Cartilage volumes calculated from the standard and fast magnetic resonance imaging sequences were similar. The median absolute percentage overestimation or underestimation of the cartilage volume with the fast sequence for 10 normal subjects was 2.5, 4.1, and 3.2% for patellar, femoral, and tibial cartilage, respectively. For eight subjects with osteoarthritis, the percentages were 4.3 and 3.9% for femoral and tibial cartilage, respectively. The fast spoiled gradient-echo sequence protocol had very high intra-rater and inter-rater reproducibility in normal subjects and in those with osteoarthritis. Because the cost of magnetic resonance imaging scanning is largely determined by scan time, the faster scanning protocol means that the cost of estimates of knee cartilage volume can be significantly reduced, thus increasing the feasibility of this methodology. PMID- 11052495 TI - Increased matrix synthesis following adenoviral transfer of a transforming growth factor beta1 gene into articular chondrocytes. AB - Monolayer cultures of lapine articular chondrocytes were transduced with first generation adenoviral vectors carrying lacZ or transforming growth factor beta1 genes under the transcriptional control of the human cytomegalovirus early promoter. High concentrations of transforming growth factor beta1 were produced by chondrocytes following transfer of the transforming growth factor beta1 gene but not the lacZ gene. Transduced chondrocytes responded to the elevated endogenous production of transforming growth factor beta1 by increasing their synthesis of proteoglycan, collagen, and noncollagenous proteins in a dose dependent fashion. The increases in collagen synthesis were not accompanied by alterations in the collagen phenotype; type-II collagen remained the predominant collagen. Transforming growth factor beta1 could not, however, rescue the collagen phenotype of cells that had undergone phenotypic modulation as a result of serial passaging. These data demonstrate that chondrocytes can be genetically manipulated to produce and respond to the potentially therapeutic cytokine transforming growth factor beta1. This technology has a number of experimental and therapeutic applications, including those related to the study and treatment of arthritis and cartilage repair. PMID- 11052496 TI - Molecular cloning, sequencing, and tissue and developmental expression of mouse cartilage oligomeric matrix protein (COMP). AB - Mouse cartilage oligomeric matrix protein cDNA was cloned and sequenced by a reverse transcription-polymerase chain reaction. The open reading frame encoded a product of 755 amino acids that shares a high degree of identity to and possesses all the characteristic molecular features of both rat and human cartilage oligomeric matrix protein. This suggests that cartilage oligomeric matrix protein is highly conserved during evolution. The clone was 83, 84, and 95% identical to human, bovine, and rat cartilage oligomeric matrix protein cDNA, respectively. In tissues from the adult mouse, cartilage oligomeric matrix protein was expressed not only in cartilage and tendon but in trachea, bone, skeletal muscle, eye, heart, and placenta as well, and no expression was found in other tissues. Immunohistology revealed that cartilage oligomeric matrix was deposited as early as 10 days post coitus in predifferentiated mouse embryo mesenchyme. It was detected in all cartilaginous tissues and in the skeletal muscles of the embryo at day 13. As development progressed, accumulation of cartilage oligomeric matrix protein was marked in the growth plate. At 19 days post coitus, it was prominently deposited in the hypertrophic zone of the growth plate, perichondrium, and periosteum and in the superficial layer of the articular cartilage surface but was absent in the more central areas of the epiphyseal cartilage. The restricted tissue distribution and expression of cartilage oligomeric matrix protein in developing as well as adult mouse tissues suggest the regulation of this protein at the transcriptional level. The findings reported herein are the first detailed characterization of the distribution of cartilage oligomeric matrix protein during early skeletal development of the mouse. PMID- 11052497 TI - Ultrastructural findings after intraarticular application of hyaluronan in a canine model of arthropathy. AB - We investigated the effect of intraarticularly applied hyaluronic acid (hyaluronan) on the cartilaginous structure of experimentally induced chondromalacia patellae in dogs. For the induction of chondromalacia, we used the Pond-Nuki technique, which involved severance and resection of the anterior cruciate ligament, as a canine model of arthropathy in 27 foxhounds (three groups of nine animals each). In a pilot study, we evaluated the effect of resection of the anterior cruciate ligament with no therapy. Patellar specimens were retrieved at 3, 6, and 12 weeks postoperatively. Subsequently, we compared a treatment group that received intraarticular injections of hyaluronan with a placebo group that received saline solution. The groups were compared at 3, 6, and 12 weeks postoperatively. Three animals from the treatment and placebo groups received five injections of hyaluronan during one of the 4-week intervals (weeks 3-6, 6-9, or 12-15). Specimens were retrieved 5 weeks after the last injection. In both groups, the uninvolved contralateral knee served as a control. The specimens were taken from the medial and lateral patellar poles. Histological analysis included light microscopy and transmission electron microscopy. The structural and ultrastructural changes were assessed qualitatively and were quantified with use of a modified Mankin score. Our results indicate that chondromalacia patellae may be induced with the Pond-Nuki technique. We found a significant reduction (p < 0.01) of cartilaginous lesions in the hyaluronan group compared with the placebo group. Our results suggest that intraarticularly applied hyaluronan is effective in delaying the degenerative process of cartilage degradation. Therefore, we conclude that the use of hyaluronan may be indicated during the early stages of chondromalacia. PMID- 11052498 TI - Influence of muscular activity on local mineralization patterns in metatarsals of the embryonic mouse. AB - This study addressed the theory that local mechanical loading may influence the development of embryonic long bones. Embryonic mouse metatarsal rudiments were cultured as whole organs, and the geometry of the primary ossification center was compared with that of rudiments that had developed in utero. The mineralization front in vivo was found to be nearly straight, whereas in vitro it acquired a more convex shape due to a slower mineralization rate at the periphery of the mineralized cylinder. A poroelastic finite element analysis was performed to calculate the local distributions of distortional strain and fluid pressure at the mineralization front in the metatarsal during loading in vivo as a result of muscle contractions in the embryonic hindlimbs. The distribution of fluid pressure from the finite element analysis could not explain the difference in mineralization shape. The most likely candidate for the difference was the distortional strain, resulting from muscle contraction, which is absent in vitro, because its value at the periphery was significantly higher than in the center of the tissue. Without external loads, the mineralization process may be considered as pre-programmed, starting at the center of the tissue and resulting in a spherical mineralization front. Strain modulates the rate of the mineralization process in vivo, resulting in the straight mineralization front. These results confirm that disturbances in muscle development are likely to produce disturbed mineralization patterns, resulting in a disordered osteogenic process. PMID- 11052499 TI - Morphometric analysis of canine skeletal muscles following experimental callus distraction according to the Ilizarov method. AB - Muscle fiber diameter and fiber-type distribution were analyzed during callus distraction. The right tibia in 24 beagles was lengthened 2.5 cm by callus distraction after osteotomy and application of a ring fixator. Distraction was started at the fifth postoperative day, at a rate of two times for 0.5 mm per day. Twelve dogs that underwent limb-lengthening and three dogs in the control group that did not undergo limb-lengthening were killed at the end of the 25-day distraction phase (group A). The remaining dogs (12 that underwent limb lengthening and three that did not) were killed after an additional consolidation period of 25 days (group B). The tibialis anterior, extensor digitorum longus, peroneus longus, and gastrocnemius muscles were removed from the right limb (which had undergone distraction) and the left control side of each animal. Crosscut cryostat sections were stained by adenosine triphosphatase at pH 4.3 and 9.4 to determine the size and distribution of types I and II fibers. Morphometric analysis of the muscle fibers was performed by a computer-assisted two-point technique. On the lengthened side, the muscles revealed marked atrophy affecting predominantly type-II fiber in the dogs in group A and affecting both fiber types in dogs in group B. Fiber density increased in both groups. In addition, fiber type grouping indicative of reinnervation was obvious in group B. Fiber-type distribution in the dogs in group B showed a shift toward type I in the tibialis anterior (p = 0.043) and extensor digitorum longus (p = 0.034) muscles and a shift toward type II in the gastrocnemius (p = 0.038). The data show that tension stress during tibial lengthening leads to atrophy of type-II fiber, reflecting disuse of muscle fiber in the distraction period as well as neurogenic atrophy followed by the reinnervation processes. Furthermore, the data are consistent with the occurrence of histoneogenesis during limb-lengthening resulting in an increase in fiber density. PMID- 11052500 TI - Primary resective shortening followed by distraction osteogenesis for limb reconstruction: a comparison with simple lengthening. AB - Resective distraction osteogenesis is a new approach to treat segmental diaphyseal bone defects by primary limb shortening and secondary distraction osteogenesis from the same site. A rabbit model was introduced to compare the bone-regeneration characteristics of this technique with those of simple lengthening procedures. Twenty-four skeletally mature New Zealand White rabbits were divided into two equal groups. In the test group, limbs were lengthened after a 10-mm segmental diaphyseal bone resection and limb shortening. In the control group, a simple subperiosteal osteotomy for limb lengthening was performed without resection. New bone formation was evaluated mechanically, radiologically, histomorphometrically, and densitometrically. Bone bridging occurred in all animals. Normalized mechanical values for the newly reconstructed tibiae demonstrated similar torsional stiffness (71+/-3.3 compared with 71+/ 8.2%; p = 0.95) and strength (64+/-5.3 compared with 68+/-7.3%; p = 0.66) in the two groups. The average normalized callus diameter was significantly greater in the test group than in the control group (p < 0.01). The remodeling index calculated from densitometry, however, showed a significantly less progressed stage of remodeling in the test group (p < 0.05). Histomorphometric analysis of the callus center supported this finding, showing significantly lower values for trabecular thickness (p < 0.05) and total bone volume (p < 0.01) in the test group. The results demonstrated the possibility of new bone formation after resection and monofocal shortening. This suggests a new therapeutic option to treat diaphyseal segmental bone defects. PMID- 11052501 TI - Pulsed electromagnetic field stimulation of MG63 osteoblast-like cells affects differentiation and local factor production. AB - Pulsed electromagnetic field stimulation has been used to promote the healing of chronic nonunions and fractures with delayed healing, but relatively little is known about its effects on osteogenic cells or the mechanisms involved. The purpose of this study was to examine the response of osteoblast-like cells to a pulsed electromagnetic field signal used clinically and to determine if the signal modulates the production of autocrine factors associated with differentiation. Confluent cultures of MG63 human osteoblast-like cells were placed between Helmholtz coils and exposed to a pulsed electromagnetic signal consisting of a burst of 20 pulses repeating at 15 Hz for 8 hours per day for 1, 2, or 4 days. Controls were cultured under identical conditions, but no signal was applied. Treated and control cultures were alternated between two comparable incubators and, therefore, between active coils; measurement of the temperature of the incubators and the culture medium indicated that application of the signal did not generate heat above the level found in the control incubator or culture medium. The pulsed electromagnetic signal caused a reduction in cell proliferation on the basis of cell number and [3H]thymidine incorporation. Cellular alkaline phosphatase-specific activity increased in the cultures exposed to the signal, with maximum effects at day 1. In contrast, enzyme activity in the cell-layer lysates, which included alkaline phosphatase-enriched extracellular matrix vesicles, continued to increase with the time of exposure to the signal. After 1 and 2 days of exposure, collagen synthesis and osteocalcin production were greater than in the control cultures. Prostaglandin E2 in the treated cultures was significantly reduced at 1 and 2 days, whereas transforming growth factor-beta1 was increased; at 4 days of treatment, however, the levels of both local factors were similar to those in the controls. The results indicate enhanced differentiation as the net effect of pulsed electromagnetic fields on osteoblasts, as evidenced by decreased proliferation and increased alkaline phosphatase-specific activity, osteocalcin synthesis, and collagen production. Pulsed electromagnetic field stimulation appears to promote the production of matrix vesicles on the basis of higher levels of alkaline phosphatase at 4 days in the cell layers than in the isolated cells, commensurate with osteogenic differentiation in response to transforming growth factor-beta1. The results indicate that osteoblasts are sensitive to pulsed electromagnetic field stimulation, which alters cell activity through changes in local factor production. PMID- 11052502 TI - Spindle-shaped cells derived from giant-cell tumor of bone support differentiation of blood monocytes to osteoclast-like cells. AB - Spindle-shaped cells were established from four giant-cell tumors of bone. When human blood monocytes were co-cultured with these cells, multinucleated giant cell formation of monocytes was induced. Intriguingly, even when a filter (pore size: 0.45 microm) was interposed between monocytes and the spindle-shaped cells, polykaryocytes also appeared. These multinucleated giant cells were positive for tartrate-resistant acid phosphatase, expressed calcitonin receptor, and showed bone-resorption activity, characteristics of osteoclast-like cells. These findings indicate that soluble factors secreted from these cells play an important role in osteoclast-like cell formation from blood monocytes. These data additionally suggest that these cells support osteoclast-like cell formation in giant-cell tumors of bone. The cells also expressed mannose receptor, fibronectin, receptor activator of nuclear factorkappaB, and several cytokine mRNAs, including interleukin-6, receptor activator of nuclear factorkappaB ligand/osteoclast differentiation factor/osteoprotegerin ligand, and macrophage colony-stimulating factor. However, all of these molecules except receptor activator of nuclear factorkappaB ligand mRNA could also be detected in control HeLa and CV-1 cells. Although the soluble receptor activator of nuclear factorkappaB ligand has not been found under physiological conditions, it is possible that it is cleaved by cellular proteases and the truncated receptor activator of nuclear factorkappaB is released from cells. Identification of the soluble factors capable of inducing osteoclast formation from blood monocytes is a pressing problem to be solved. PMID- 11052503 TI - Cytokines associated with the pathophysiology of aggressive fibromatosis. AB - The rare benign extra-abdominal desmoid tumor is characterized by aggressive invasion of normal tissue. Treatment is complicated by its recurrence, invasiveness, and persistence. The etiology is unknown and the pathophysiology is obscure. Because of exuberant fibroblastic proliferation with collagenous tissue being the primary tissue component, this desmoid tumor has been compared with keloids arising from excessive scar formation in healing wounds. Numerous cytokines are associated with signaling for growth and maintenance of mesenchymal cells. Altered expression of these proteins is associated with many pathologic conditions. It has been proposed that the enhanced expression of platelet-derived growth factor and its receptor characterize desmoid tumors. We tested the hypothesis that the exuberant fibrosis of desmoid tumors may have resulted from the initiation of the cascade of molecular events producing increased expression of cytokines. We used immunohistochemical analysis of cytokines in desmoid tumors compared with keloids and skin to localize the expression of cytokines. The results showed localized increased expression of the cytokines epidermal growth factor, transforming growth factor-beta, tumor necrosis factor-alpha, vascular endothelial growth factor, interleukin-1beta, and interleukin-6 in the endothelial cells of blood vessels in the tumors. Production of tumor necrosis factor-alpha and interleukin-1beta in tumor tissue was increased, but we did not find increased expression of platelet-derived growth factor. We concluded that the increased expression of cytokines associated with angiogenesis usually found in wound healing and invasive tumors may contribute to the pathophysiology of the desmoid tumor. PMID- 11052504 TI - Adaptation of post-traumatic angiogenesis in the rabbit knee by apposition of torn ligament ends. AB - Apposition of torn ligament ends has been shown to have a beneficial effect on healing of the medial collateral ligament; however, the mechanism underlying this improved recovery is unclear. Excessive post-traumatic angiogenesis, an inherent component of soft-tissue regeneration, may be functionally detrimental in relatively hypovascular tissues such as ligaments. The present study therefore examined the relationship between contact of transected ligament ends and vascular remodeling. Female New Zealand White rabbits were subjected to a gap injury, Z-plasty apposition, or sham operation to the midsubstance of the medial collateral ligament. Six weeks after treatment, the volume of vessels supplying the healing zone of the medial collateral ligament, as well as the ipsilateral lateral collateral ligament, posterior cruciate ligament. menisci, and medial capsule, was quantified by carmine red vascular casting. The volume of vessels supplying the neoligamentous scar formed by gap injury to the medial collateral ligament was found to be twice that of ligaments that had undergone the sham operation, and lateral collateral ligament and meniscal vascularity was also augmented in the injured joint. The medial collateral ligaments that underwent Z plasty apposition exhibited a level of vascularity comparable with that of the control ligaments that had undergone the sham procedure, whereas meniscal and lateral collateral ligament vascularities remained elevated in this group. Capsular and posterior cruciate ligament vascularities were unaffected by gap injury or Z-plasty to the ipsilateral medial collateral ligament. These findings indicate that injury to the medial collateral ligament not only stimulates angiogenesis in the healing ligament, but other ipsilateral soft tissues also undergo vascular remodeling. Furthermore, apposition of an injured medial collateral ligament modifies these pro-angiogenic events, and this may partly explain why contact of torn ligament ends is beneficial for post-traumatic recovery of an injured joint. PMID- 11052505 TI - Poroelastic properties of bovine vertebral trabecular bone. AB - The theory of poroelasticity has been used to study bone mechanics without directly measuring poroelastic properties. In this study, we developed an experimental protocol and measured the poroelastic properties of bovine vertebral trabecular bone. Mean (+/-SD) values for drained shear modulus, drained Poisson's ratio, undrained Poisson's ratio, Skempton's coefficient, and permeability coefficient were, respectively, 90.85 (+/-59.59) MPa and 0.242 (+/-0.099), 0.399 (+/-0.083), 0.851 (+/-0.144), and 16.31 (+/-8.02) x 10(-8) m2/Pa/sec, respectively. The experimental protocol can be used generally for the measurement of poroelastic properties of bone when cylindrical specimens are available. Measured poroelastic properties can be used directly or converted to Biot's coefficient and modulus, without assuming the incompressibility of solid and fluid constituents, for the poroelastic modeling of bone. PMID- 11052506 TI - Effect of in vitro testing over extended periods on the low-load mechanical behaviour of dense connective tissues. AB - Many biomechanical studies are performed on dense connective tissues in the laboratory over a substantial time period; however, the effect of the in vitro testing environment on the cyclic load-relaxation behaviour of these structures is not well established. This study evaluated the effect of long duration of testing on ligament viscoelastic behaviour, using the rabbit femur-medial collateral ligament-tibia complex as a model. Dissected rabbit knee joints were mounted on a materials testing machine, and isolated ligament complexes were cycled at a frequency of one cycle/min to a fixed displacement of 0.7 mm for an 18-hour period. After an initial period of exponential load relaxation, the cyclic peak loads slowly decreased over the 18-hour period. The average decrease in the cyclic peak load between 0.5 and 18.0 hours was 0.26% (of the original peak load) per hour (r2 = 0.934), or a total of 8.6+/-4.6% over this period (p < 0.0001). Thus, low-load testing of dense connective tissues in the laboratory over extended periods significantly alters their biomechanical behaviour, and these changes should be considered in long-term laboratory-based studies of dense connective tissues. PMID- 11052507 TI - Custom-made AO unreamed interlocked tibial nails. PMID- 11052508 TI - In vivo analysis of microcirculation following closed soft tissue injury. PMID- 11052509 TI - Treatment of primary colon cancer with peritoneal carcinomatosis: comparison of concomitant vs. delayed management. AB - PURPOSE: The initial dissemination of colon cancer occurs through three routes: the lymphatics, the portal blood, and the peritoneal surfaces. Although lymphatic and hematogenous metastases indicate an aggressive disease process, it is possible that dissemination to peritoneal surfaces may be only superficial contamination of the parietal and visceral peritoneum that may be treatable for cure. Unfortunately, surgery may have an adverse impact on peritoneal surface dissemination. Surgical interventions may convert a superficial process into an invasive condition with a greatly reduced prognosis. This study was conducted to test this hypothesis by the use of data prospectively recorded from patients treated for peritoneal carcinomatosis concomitant with resection of the primary colon cancer or treated for carcinomatosis after disease recurrence prompted referral. METHODS: Our first group of patients had definitive treatment of carcinomatosis simultaneous with resection of the primary colon cancer. They had cytoreductive surgery including peritonectomy procedures followed by heated intraoperative intraperitoneal chemotherapy with mitomycin C plus early postoperative intraperitoneal 5-fluorouracil. The comparison group was treated with a colon resection at an outside hospital and then later referred to us with progressive disease for treatment. The major difference between the groups is the timing of the definitive treatment of carcinomatosis. These patients were also studied by use of the completeness of the cytoreduction score and the peritoneal cancer index as prognostic indicators. Survival was the end point for all the analysis. RESULTS: Of 104 patients with peritoneal carcinomatosis from colon or rectal adenocarcinoma, five patients (4.8 percent) had definitive treatment of the peritoneal surface spread of the cancer concomitant with resection of the primary lesion. Median survival for these patients has not been reached and their five-year survival rate is 100 percent. The remainder of the patients (n = 99) were referred for local and regional recurrence after their primary cancer had been removed and there was progression of carcinomatosis. Forty-four patients (42.3 percent) had a complete cytoreduction resulting in a 24-month median survival and a 30 percent five-year survival (P < 0.0001). The other 55 patients (52.9 percent) had an incomplete cytoreduction resulting in a 12-month median survival and a 0 percent five-year survival (P < 0.0001). Patients with a peritoneal cancer index of 10 or less had a 48-month median survival and a 50 percent five-year survival rate. Patients with a peritoneal cancer index between 11 and 20 had a 24-month median survival and a 20 percent five-year survival rate (P < 0.0001). Patients with a peritoneal cancer index of more than 20 had a 12 month median survival and a 0 percent five-year survival (P < 0.0001). CONCLUSIONS: In patients with peritoneal seeding occurring at the time of resection of the primary malignancy, peritonectomy procedures and perioperative intraperitoneal chemotherapy should be performed concomitantly. By use of a quantitative scoring system, the mass of cancer present in the abdomen and pelvis at the time of treatment of carcinomatosis correlated directly with survival. Aggressive treatment of patients with peritoneal carcinomatosis requires consideration in the management of colorectal cancer. PMID- 11052510 TI - Anorectal nomenclature: fundamental terminology. AB - There is lack of agreement on the definitions of the anal canal and its parts. To facilitate mutual understanding and communication between workers, it is highly desirable that a set of agreed-upon definitions be developed. The development of the different definitions and their purposes is followed and they are analyzed. As a basis for discussion, a possible revised entry for the next edition of Terminologia Anatomica is presented. Draft definitions of the terms in the entry are provided. Practitioners are invited to become involved in the process of developing agreement on definitions by providing comments and criticism. PMID- 11052511 TI - Topical diltiazem and bethanechol decrease anal sphincter pressure and heal anal fissures without side effects. AB - PURPOSE: Topical glyceryl trinitrate heals anal fissures, but a majority of patients experience headache. Topical gels of the calcium channel blocker diltiazem and the cholinomimetic bethanechol significantly lower anal sphincter pressure in volunteers. This study investigated the use of these two new pharmacologic agents in the treatment of patients with chronic anal fissure. METHODS: Two studies were conducted, each involving 15 patients with chronic anal fissure. In each study patients underwent anal manometry and laser doppler flowmetry before treatment. They were treated with either 2 percent diltiazem gel or 0.1 percent bethanechol gel three times daily for eight weeks. Assessment every two weeks was by clinical examination, repeat anal manometry, and laser doppler flowmetry. Daily pain was assessed by linear analog charts. RESULTS: Fissures healed in 10 of 15 (67 percent) patients treated with 2 percent diltiazem gel and in 9 (60 percent) patients treated with 0.1 percent bethanechol gel. There was no significant difference in the pretreatment maximum resting sphincter pressure (MRP) between responders and nonresponders in either group. There was significant reduction in the pain score after treatment with diltiazem (P = 0.002) and bethanechol (P = 0.005) compared with that before treatment. MRP was significantly lower after diltiazem (P = 0.0001) and bethanechol (P = 0.02) compared with pretreatment MRP. No headaches or side effects were reported. CONCLUSIONS: Both topical diltiazem and bethanechol substantially reduce anal sphincter pressure and achieve fissure healing to a similar degree reported with topical nitrates, but without side effects. PMID- 11052512 TI - Acute left colonic diverticulitis--compared performance of computed tomography and water-soluble contrast enema: prospective evaluation of 420 patients. AB - PURPOSE: The most valuable radiologic examination to be done initially when acute left colonic diverticulitis is suspected is still a matter of controversy. This study compares the performance between water-soluble contrast enema and computed tomography. METHODS: From 1986 to 1997, all patients admitted in our emergency center with clinically suspected left-colonic diverticulitis had a contrast enema and a computed tomography within 72 hours of their admission, unless clinical findings required immediate laparotomy. They were prospectively included in the study if one or both radiologic examinations showed signs of acute diverticulitis or diverticulitis was surgically removed and histologically proven or both. Diverticulitis was considered moderate when computed tomography showed localized thickening of the colonic wall (5 mm or more) and inflammation of pericolic fat and contrast enema showed segmental lumen narrowing and tethered mucosa; it was considered severe when abscess or extraluminal air or contrast or all three were observed on computed tomography and when one or both of the last two signs were seen on contrast enema. Of 542 patients, 420 who had both computed tomography and contrast enema entered the study. RESULTS: The performance of computed tomography was significantly superior to contrast enema in terms of sensitivity (98 vs. 92 percent; P = 0.01), which was calculated from patients who had their colon removed and whose diverticulitis was histologically proven, and in the evaluation of the severity of the inflammation (26 vs. 9 percent; P = 0.02). Moreover, of 69 patients who had an associated abscess seen on computed tomography, only 20 (29 percent) had indirect signs of this complication on contrast enema. CONCLUSIONS: In the diagnostic evaluation of acute left-colonic diverticulitis, computed tomography should be preferred to contrast enema as the initial radiologic examination because of its statistically significant superiority in sensitivity and for its significantly better performance in the detection of severe infection, especially when an abscess is associated with the disease. PMID- 11052513 TI - The effect of pelvic ileal reservoir volume and antiperistaltic reflux on emptying efficiency. AB - PURPOSE: The emptying efficiency of four different designs of pelvic ileal reservoir was compared using two different techniques of measurement. METHOD: Thirty-four patients were studied one year after restorative proctocolectomy. In each the ileal reservoir was filled with methyl cellulose paste labeled with 51chromium-chromate and technetium Tc 99m-diethylenetriamine pentaacetic acid. Percentage evacuation was calculated from 1) the difference in 51chromium activity between the recovered effluent and the total paste administered and 2) gamma camera measurements of technetium Tc 99m-diethylenetriamine pentaacetic acid activity within the ileal reservoir before and after evacuation. RESULTS: Median evacuation using the 51chromium method was 84, 90, 70, and 75 percent for the W40, W30, J40, and J30 reservoirs respectively. The results were not significantly different from those obtained using the gamma camera: 83, 87, 67, and 71 percent (P = not significant). Patients with either type of W reservoir evacuate isotope-labeled paste more efficiently than patients with J40 reservoirs (P < 0.05 and P < 0.001, respectively) but not J30 reservoirs (P = not significant). However, if the actual volume of paste evacuated during a visit to the lavatory is measured, it is greatest for J40 reservoirs (median, 300 ml compared with 258 ml for W40, 289 ml for W30, and 268 ml for J30; P = not significant). CONCLUSIONS: Gamma camera measurement of ileal reservoir emptying is as accurate as our previous standard technique and provides a qualitative record of pouch evacuation, which may reveal reasons for inefficient emptying. The gamma camera images reveal that the difference in emptying percentage between W and J pouches is because of reflux of paste into the afferent ileum occurring more frequently in J pouches than in W pouches. The effect of this phenomenon on emptying is more than compensated for by the increase in reservoir capacity created by the reflux. PMID- 11052514 TI - How can the assessment of fistula-inano be improved? AB - PURPOSE: Fistula-in-ano anatomy and its relationship with anal sphincters are important factors influencing the results of surgical management. Preoperative definition of fistulous track(s) and the internal opening play a primary role in minimizing iatrogenic damage to the sphincters and recurrence of the fistula. METHODS: Physical examination and endoanal ultrasound (performed with a 10 MHz endoprobe), either conventionally or with an injection of hydrogen peroxide, were performed in 26 consecutive patients. Results were matched with surgical features to establish their accuracy in preoperative fistula-in-ano assessment. RESULTS: Accuracy rates of clinical examination endoanal ultrasound, and hydrogen peroxide enhanced ultrasound were 65.4, 50, and 76.9 percent for primary tracks, 73.1, 65.4, and 88.5 percent for secondary tracks, and 80.8, 80.8, and 92.3 percent for horseshoe extensions, respectively. Compared with physical examination and endoanal ultrasound, accuracy of hydrogen peroxide-enhanced ultrasound was higher for transsphincteric and intersphincteric primary tracks and horseshoe extensions. Both endoanal ultrasound and hydrogen peroxide-enhanced ultrasound displayed a significantly higher accuracy in detecting the internal openings (53.8 and 53.8 percent, respectively) compared with clinical evaluation (23.1 percent; P = 0.027). CONCLUSIONS: Our data suggest that hydrogen peroxide enhanced ultrasound can be very reliable and useful in the definition of fistula anatomy, its relationship with anal sphincters, and, hence, surgical strategy. It also improves identification of secondary extensions, particularly horseshoe tracks. This method, besides being safe, economic and reputable, both preoperatively and postoperatively, could be helpful in checking operative results and recurrence. PMID- 11052515 TI - Recent trends in diverticulosis of the right colon in Japan: retrospective review in a regional hospital. AB - PURPOSE: Diverticulosis of the right colon has been increasing in the Far East; however, a considerable proportion of these patients includes cases of solitary right-sided diverticular disease. This study aimed to determine whether the incidence of such solitary diverticula (defined as 1 or 2 diverticula in this study) and multiple (3 or more) diverticula of the right colon is increasing in Japan. METHODS: A total of 13,947 consecutive barium enema examinations, performed in the period from 1982 to 1997, were reviewed. Changes in the frequency (detection rate) and number of diverticula across time and with aging of three types of diverticula, right-sided, left-sided, and bilateral, were investigated, with special interest in those patients with one or two diverticula of the right colon. RESULTS: Right-sided and bilateral diverticula have increased in frequency across time; however, left-sided diverticula have not. Patients with one or two diverticula in the right colon of right-sided disease, unexpectedly, have increased across time in both genders, and patients with three or more diverticula in the right colon of right-sided disease have shown an increase in males. The number of diverticula of the right colon showed no increase across time or with aging. CONCLUSIONS: Diverticulosis of the right colon, both solitary and multiple, has been increasing steadily in Japan; therefore, diverticulitis and bleeding diverticula of the right colon may continue to increase. Diverticula of the right colon might be an acquired disease and self-limiting in development, because the frequency did not increase substantially in the elderly and because the number changed little across time and with aging. PMID- 11052516 TI - Neuroanatomy of the pelvis: implications for colonic and rectal resection. AB - PURPOSE: Urinary dysfunction remains a common complication of radical pelvic surgery, particularly after abdominoperineal resection. In treating rectal carcinoma, the extent of primary resection and lymphadenectomy are major determinants in the degree of postoperative urologic morbidity. METHODS: Twelve male and eight female hemipelves from fresh cadavers were dissected with reference to the neuroanatomy of the lower genitourinary tract. These cadavers were dissected within twelve hours of thaw from frozen state. The cadavers were hemisected at the level of the sacral promontory for better exposure of neural trunks and vascular structures leading into the pelvis. These structures were followed down sequentially into the true pelvis, using magnified dissection under operating microscope or loupe dissection or both. RESULTS: Coordinated lower urinary tract function relies on both autonomic and somatic nerve activity. Emanating from the inferior hypogastric plexus, the pelvic nerve supplies sympathetic and parasympathetic innervation to the pelvic viscera. The course of the pelvic nerve is as follows: 1) from the inferior hypogastric plexus, it has multiple branches forming a web-like complex within the endopelvic fascial sleeve, some of which innervate the bladder detrusor; 2) a main branch traveling inferolateral to the rectum remains deep to the fascia of the levator ani muscle and courses to the external urinary sphincter; 3) at the level of the prostatic apex (or bladder neck in females), this pelvic nerve branch sends direct branches to the urinary sphincter. The pudendal nerve traverses the pelvis in the pudendal canal, and before leaving the pelvis to enter the perineum, it gives an intrapelvic branch that courses alongside the ischium to enter the external urinary sphincter. In the ischiorectal fossa, terminal branches of the pudendal nerve (i. e., perineal nerve) can be seen inserting into the urinary sphincter. CONCLUSIONS: Urinary retention and urinary incontinence represent two distinct urologic complications after abdominoperineal resection. Injury to detrusor branches of the pelvic nerve can cause detrusor denervation and urinary retention. In addition, injury to intrapelvic branches of the pelvic and pudendal nerves to the urinary sphincter can result in intrinsic sphincter deficiency and urinary incontinence. A better understanding of the neuroanatomy of the lower genitourinary tract can give a physiologic basis for clinical findings of postoperative voiding dysfunction and may help the surgeon refine surgical technique by more precisely determining resection limits to minimize urologic complications. PMID- 11052517 TI - Functional outcomes in patients with mucosal ulcerative colitis after ileal pouch anal anastomosis by the double stapling technique: is there a relation to tissue type? AB - PURPOSE: The aim of this study was to evaluate any differences in functional outcome in patients with mucosal ulcerative colitis after restorative proctocolectomy and ileal pouch-anal anastomosis with use of the double stapling technique relative to the type of tissue in the stapled doughnut. METHODS: Between September 1988 and June 1997, the pathology of all patients with mucosal ulcerative colitis who underwent ileal pouch-anal anastomosis with use of the double stapling technique were reviewed. Information was obtained regarding the tissue types in the distal tissue rings (doughnuts) obtained from the stapled ileal pouchanal anastomosis. The level of anastomosis was classified according to the type of tissue in the distal doughnut: Group I- patients in whom the anal transitional zone was removed and the distal doughnut included squamous epithelium or transitional epithelium and Group II- patients in whom the anal transitional zone was preserved because the distal doughnut revealed only columnar epithelium. Functional outcomes were assessed and compared by detailed questionnaires mailed to all patients at least one year after ileal pouch-anal anastomosis surgery. RESULTS: Distal doughnuts were obtained from the stapled ileal pouch-anal anastomosis in 222 patients with mucosal ulcerative colitis. Follow-up data at a mean of 38 (range, 12-132) months were obtained in 138 (62.2 percent) patients, including 72 males, with a mean age of 46.9 (range, 13-79) years. Group I consisted of 40 patients (29 percent; 35 (25.4 percent) who had squamous epithelium and 5 (3.6 percent) who had transitional epithelium in the distal tissue rings). Group II consisted of 98 patients (71 percent) with columnar epithelium in the distal tissue rings. Age at diagnosis and operation, duration of disease, length of follow-up, and stage of pouch surgery were similar in the two groups. Incontinence scores, frequency of bowel movement, use of a protective pad, discrimination between gas and stool, use of antidiarrheals, life style alteration, and patient satisfaction showed similar functional results between the two groups. CONCLUSIONS: The tissue type in the stapler distal doughnut did not greatly influence functional outcome. Failure to identify a relationship may attest to the variable height and composition of the anal transitional zone. PMID- 11052518 TI - An increased rectal maximum tolerable volume and long anal canal are associated with poor short-term response to biofeedback therapy for patients with anismus with decreased bowel frequency and normal colonic transit time. AB - PURPOSE: Biofeedback is an effective therapy for a majority of patients with anismus. However, a significant proportion of patients still failed to respond to biofeedback, and little has been known about the factors that predict response to biofeedback. We evaluated the factors associated with poor response to biofeedback. METHODS: Biofeedback therapy was offered to 45 patients with anismus with decreased bowel frequency (less than three times per week) and normal colonic transit time. Any differences in demographics, symptoms, and parameters of anorectal physiologic tests were sought between responders (in whom bowel frequency increased up to three times or more per week after biofeedback) and nonresponders (in whom bowel frequency remained less than three times per week). RESULTS: Thirty-one patients (68.9 percent) responded to biofeedback and 14 patients (31.1 percent) did not. Anal canal length was longer in nonresponders than in responders (4.53 +/- 0.5 vs. 4.08 +/- 0.56 cm; P = 0.02), and rectal maximum tolerable volume was larger in nonresponders than in responders. (361 +/- 87 vs. 302 +/- 69 ml; P = 0.02). Anal canal length and rectal maximum tolerable volume showed significant differences between responders and nonresponders on multivariate analysis (P = 0.027 and P = 0.034, respectively). CONCLUSIONS: This study showed that a long anal canal and increased rectal maximum tolerable volume are associated with poor short-term response to biofeedback for patients with anismus with decreased bowel frequency and normal colonic transit time. PMID- 11052519 TI - Dietary habits and right-sided colonic diverticulosis. AB - PURPOSE: In Asian populations, there is a high prevalence of right-sided colonic diverticulosis, the cause of which is uncertain. It is suspected that dietary habits may interact with a congenital predilection to cause this condition. To evaluate the relationship between long-term dietary habits and the prevalence of right-sided diverticulosis in the general population, we performed a retrospective case-control study. METHODS: We reviewed the records of 3,105 screening colonoscopies performed on healthy, asymptomatic adults. All cases of right-sided diverticulosis were selected, and a similar number of gender-matched and age-matched controls with negative colonoscopies were randomly sampled from the same cohort. All case and control subjects were interviewed by a single blinded nurse to establish their dietary habits during the past decade, in addition to other demographic characteristics. Based on consumption frequency, they were assigned to one of three diet classes for each of three food categories of interest: meat, vegetable, and fruit products. Staple foods such as rice were not included. Odds ratios were then calculated using multivariate conditional logistic regression and tests for trend were performed. RESULTS: A total of 86 cases of right-sided diverticulosis were included, whereas 106 controls were randomly selected. There was a marked association between meat consumption frequency and right-sided diverticulosis, with a trend P value of <0.01 and an odds ratio of 24.81 between the most and least frequent consumers of meat products. CONCLUSIONS: The prevalence of right-sided diverticulosis is strongly positively associated with past meat consumption frequency. There is no association with vegetable or fruit consumption frequency, laxative use, supplemental fiber intake, smoking, or family history. PMID- 11052520 TI - Linear discriminant analysis of symptoms in patients with chronic constipation: validation of a new scoring system (KESS). AB - PURPOSE: The aim of this study was to devise a symptom scoring system to assist in diagnosing constipation and in discriminating among pathophysiologic subgroups. METHOD: A structured symptom scoring questionnaire (11 questions) was completed by 71 chronically constipated patients and by 20 asymptomatic controls. The symptom score was correlated with a previously validated constipation score (Cleveland Clinic Score). All patients underwent colonic transit studies, standard anorectal physiology testing, and evacuation proctography. On the basis of these investigations alone, an observer blinded to the questionnaire results allocated patients to one of three pathophysiologic subgroups: slow-transit constipation, rectal evacuatory disorder, or mixed (slow-transit constipation and rectal evacuatory disorder). Linear discriminant analysis was used to assess the ability of different questionnaire symptoms to discriminate among these subgroups. RESULTS: Total symptom score was strongly correlated with the Cleveland Clinic Score (r = 0.9). The median total score in constipated patients was 20 (range, 11-35) compared with a median of 2 in controls (range, 0-6). Discriminant analysis using cross validation estimated that pathophysiology could be predicted correctly for 55 percent (95 percent confidence interval = 43-67 percent) of patients using just five symptoms. The discriminant function rarely misclassified patients with rectal evacuatory disorder as slow-transit constipation and vice versa, but could not effectively discriminate between patients with single and mixed pathologies. CONCLUSION: This new scoring system is a valid technique to assist in the diagnosis of constipation and is the first study using appropriate statistical methodology to demonstrate a discriminatory ability of multiple symptoms in constipation. At present, symptom analysis does not adequately differentiate major pathophysiologic subgroups for use in clinical practice. PMID- 11052521 TI - Enteral inulin does not affect epithelial gene expression and cell turnover within the ileoanal pouch. AB - PURPOSE: This study evaluates the effects of enteral inulin on ileoanal pouch functioning by studying epithelial gene expression, cell turnover, and mucosal morphology. METHODS: Twenty patients with an ileoanal pouch received 24 g of inulin daily for three weeks, then a four-week wash-out period, and a placebo for three weeks. In this randomized, double-blind, crossover study, biopsy specimens of pouch mucosa were taken after each test period. Mucosal morphology, inflammation, epithelial proliferation, and cell death were assessed histologically. Expressions of proapoptotic and antiapoptotic regulators, intestinal fatty acid-binding protein, and mucin were quantified by Western blotting or enzyme-linked immunosorbent assay. The number of intestinal fatty acid-binding protein expressing cells was histologically assessed and a high iron diamine/Alcian blue staining was performed to discriminate between sulfated and nonsulfated acidic mucins. RESULTS: Inulin supplementation neither altered mucosal morphology nor influenced inflammation, epithelial cell proliferation, or cell death. The ratio between the proapoptotic and antiapoptotic regulators did not change after inulin supplementation. The number of intestinal fatty acid binding protein-producing enterocytes and the intestinal fatty acid-binding protein expression level increased after inulin treatment, but did not reach statistical significance. The intestinal fatty acidbinding protein expression level correlated with the Pouchitis Disease Activity Index, which was at the brink of significance (P = 0.06). Mucin expression and the ratio between sulfated and nonsulfated acidic mucins were not altered by inulin supplementation. CONCLUSION: In this prospective study, inulin supplementation did not significantly alter pouch mucosal functioning because neither epithelial homeostasis nor epithelial gene expression was significantly altered by enteral inulin. PMID- 11052522 TI - Use of hyperbaric oxygen chamber in the management of radiation-related complications of the anorectal region: report of two cases and review of the literature. AB - PURPOSE: This article was undertaken to present two cases of nonhealing ulcers that occurred after primary radiation therapy and local excision of suspected residual or recurrent anal carcinomas. Both patients responded favorably to hyperbaric chamber treatment. Review of the literature is discussed, including cause, clinical presentation, diagnosis, and options for management of radiation related complications in the anorectal region and use of hyperbaric oxygen treatment in colorectal surgery. METHODS: The cases of two patients with recurrent or residual anal carcinomas were reviewed. Objective clinical, laboratory test, and intraoperative findings were implemented to define this pathologic entity precisely, results of its treatment, and management of radiation-related complications. RESULTS: The study shows clinical effectiveness of hyperbaric chamber treatment for nonhealing wounds in the previously radiated anorectal region. The refractory wounds of both our patients healed. The patients were rendered free of symptoms. CONCLUSIONS: Substantial pathologic changes in the irradiated tissues leading occasionally to nonhealing radiation proctitis are relatively infrequent consequences of radiation therapy for pelvic malignancies. Excisional and incisional biopsies of the radiation-injured tissues result in chronic ulcers accompanied by debilitating symptoms. Hyperbaric chamber treatment seemed to be a very effective means of therapy of radiation proctitis and nonhealing wounds in the involved anorectal region after conventional therapy had failed. PMID- 11052523 TI - Rectosigmoidal adenomatous polyposis: a novel entity of polyposis? Report of a case. AB - PURPOSE: We report a patient with rectosigmoidal adenomatous polyposis. METHODS: A 57-year-old male presented with a submucosally invasive well-differentiated adenocarcinoma in the rectum and approximately 100 adenomatous polyps with an extremely unusual distribution limited exclusively to the rectum and sigmoid colon. RESULTS: There was no family history of colorectal disease or any related disorders. No extracolonic manifestations were found. Because this case was considered to be a discriminative phenotype of familial adenomatous polyposis, DNA from a peripheral sample of whole blood was screened for APC germline mutation by a combination of protein truncation test and single stranded conformation polymorphism, but no mutation was found. CONCLUSION: This patient may have a novel entity of adenomatous polyposis with a peculiar distribution. It may be caused by some genetic alteration other than APC mutation. PMID- 11052524 TI - Granular cell tumor--a unique neoplasm of the internal anal sphincter: report of a case. AB - Granular cell tumors are rare, invariably benign, and often solitary tumors, which infrequently involve the gastrointestinal tract. We report the unique presentation of a granular cell tumor of the internal anal sphincter in a 75 year old female. The tumor was detected during investigation of new rectal bleeding and was excised using a transanal approach and sphincter repair. At five-year follow-up the patient reported normal continence to stool and flatus and demonstrated no evidence of tumor recurrence. Immunohistochemical studies cite the Schwann neural cell as the origin of the granular cell tumor. Cytoplasmic features include acidophilic, p-aminosalicylic acid-positive, diastaseresistant granules. Granular cell tumors may be located anywhere in the body, but anorectal involvement is rare. In our own search of the world literature, no other cases were reported specifically to involve the anal sphincter. Granular cell tumors are usually detected incidentally but may be symptomatic, especially when the anorectal region is involved. Symptoms include perianal discomfort and bleeding. Adequate local excision is effective for both diagnosis and treatment of anorectal granular cell tumors. Careful follow-up should be performed after excision because of the risk of recurrence. PMID- 11052525 TI - Colonic "coloplasty": novel technique to enhance low colorectal or coloanal anastomosis. AB - After low colorectal or coloanal anastomosis, bowel dysfunction may exist. A colonic J-pouch has been proposed to reduce bowel dysfunction. We present an alternative technique to augment the reservoir function of the neorectum and reduce bowel dysfunction. PMID- 11052526 TI - Simple technique for laparoscopic paracolostomy hernia repair. AB - PURPOSE: The aim of this study was to describe a simple, minimally invasive, and effective technique for repair of paracolostomy hernia. METHODS: Mesh is fastened laparoscopically over the colostomy and hernial defect with wide overlap, without dissecting out the sac or repairing the defect, while ensuring enough room for the colon to prevent obstruction at the level of the mesh. RESULTS: The technique has been used successfully in four patients, with follow-up of 2 to 12 months. Operating time and length of stay were short, and there was no recurrence or prolapse of the colostomy. CONCLUSIONS: This seems to be an effective, simple, and minimally invasive technique for repairing a difficult problem. Although the number of cases is small and the follow-up has been short, the technique mimics that used in massive ventral hernia repair with good results. PMID- 11052527 TI - Neostigmine treatment for acute colonic pseudo-obstruction. PMID- 11052528 TI - Anorectal mobilization or intersphincteric resection... PMID- 11052529 TI - Ultrasonic dissection for pseudomyxoma peritonei: debulking, not cytoreduction. PMID- 11052530 TI - Wings and bristles: character specificity of the asymmetry phenotype in insecticide-resistant strains of Lucilia cuprina. AB - We investigated the hypothesis that observed higher levels of asymmetry displayed by insecticide-resistance genotypes of Lucilia cuprina are restricted to bristle characters, due to the action of resistance genes in bristle cell development, rather than through the disruption of genomic coadaptation. We compared the level of asymmetry of three bristle characters and three wing characters in non modified and modified-resistance genotypes. Consistent with previous studies, resistance genotypes displayed greater levels of bristle asymmetry than either susceptible or modified genotypes. However, there were no differences among genotypes for any of the wing characters. To confirm that this result is attributable to the action of the resistance and modifier genes themselves, we also examined the responses of both bristle and wing characters to the more general developmental stress of extreme temperature. Sub-optimal temperature was shown to increase both bristle and wing asymmetry, suggesting that there are no underlying differences between the two character types which could, of themselves, explain the differential response observed in the resistance genotypes. PMID- 11052531 TI - Chromosome fission associated with growth of ribosomal DNA in Neodiprion abietis (Hymenoptera: Diprionidae). AB - The haploid complement consists of seven metacentric chromosomes in most diprionid species but has evolved to n = 8 by fission in Neodiprion abietis. This fission generated a small telocentric chromosome and a large pseudoacrocentric chromosome with a short arm carrying a satellite. In situ hybridization indicated that the location of the rRNA gene cluster corresponds to the whole short arm. This suggests that (i) the breaking point was located close to an rRNA gene cluster, and (ii) fission was associated with growth of rDNA. These results suggest rDNA as a preferential breaking point but with a role in the healing of naked chromosome ends. PMID- 11052532 TI - Multiple origins of XY female mice (genus Akodon): phylogenetic and chromosomal evidence. AB - Despite the diversity in sex determination across organisms, theory predicts that the evolution of XY females is rare in mammals due to fitness consequences associated with infertility or the loss of YY zygotes. We investigated this hypothesis from a phylogenetic perspective by examining the inter- and intraspecific distribution of Y chromosomes in males and females (XY females) in South American field mice (Akodon). We found that XY females occurred at appreciable frequencies (10-66%) in at least eight Akodon species, raising the possibility that this system of sex determination has arisen multiple times independently. To determine the number of origins of XY females in Akodon, we constructed a molecular phylogeny of 16 species of Akodon based on mitochondrial DNA control region sequences. Both parsimony and maximum-likelihood reconstruction of ancestral states suggest that multiple steps (gains or losses of XY females) best explain the evolution of XY females, but do not clearly differentiate between single and multiple origins. We then directly compared functional and non-functional Y chromosomes in six species by Southern blot analysis. We found that male and female Y chromosome restriction fragment length polymorphism patterns were identical within species, but always differed between species, providing evidence that XY females arose at least six times within the Akodon lineage. To our knowledge, this pattern in Akodon is the first documentation of a novel sex-determining system arising multiple times within a tight clade of mammals. In addition, this system provides a clear test of the accuracy of phylogenetic methods to reconstruct ancestral states. PMID- 11052533 TI - Molecular systematics of European Hyalodaphnia: the role of contemporary hybridization in ancient species. AB - We examined phylogenetic relationships among Daphnia using mitochondrial DNA (mtDNA) sequences from the small subunit ribosomal RNA (12S), cytochrome c oxidase subunit I and nuclear DNA sequences from the first and second internal transcribed spacer representing 1612 base positions. Phylogenetic analyses using several species of the three main Daphnia subgenera, Ctenodaphnia, Hyalodaphnia and Daphnia, revealed that the Hyalodaphnia are a monophyletic sister group of the Daphnia. Most Hyalodaphnia species occur on one continent, whereas only three are found in North America and Europe. Endemicity of species is associated with variation in thermal tolerance and habitat differentiation. Although many species of the Hyalodaphnia are known to hybridize in nature, mtDNA divergence is relatively high ca. 9%) compared to other hybridizing arthropods (ca. 3%). Reproductive isolation in Daphnia seems to evolve significantly slower than genetic isolation. We related these findings to what is known about the ecology and genetics of Daphnia in order to better understand the evolutionary diversification of lineages. The relationship of these data to phylogenetic patterns is discussed in the context of speciation processes in Daphnia. PMID- 11052534 TI - Geographic range size and evolutionary age in birds. AB - Together with patterns of speciation and extinction, post-speciation transformations in the range sizes of individual species determine the form of contemporary species range-size distributions. However, the methodological problems associated with tracking the dynamics of a species' range size over evolutionary time have precluded direct study of such range-size transformations, although indirect evidence has led to several models being proposed describing the form that they might take. Here, we use independently derived molecular data to estimate ages of species in six monophyletic groups of birds, and examine the relationship between species age and global geographic range size. We present strong evidence that avian range sizes are not static over evolutionary time. In addition, it seems that, with the regular exception of certain taxa (for example island endemics and some threatened species), range-size transformations are non random in birds. In general, range sizes appear to expand relatively rapidly post speciation; subsequently; and perhaps more gradually, they then decline as species age. We discuss these results with reference to the various models of range-size dynamics that have been proposed. PMID- 11052535 TI - Visibility of the environmental noise modulating population dynamics. AB - Characterizing population fluctuations and their causes is a major theme in population ecology. The debate is on the relative merits of density-dependent and density-independent effects. One paradigm (revived by the research on global warming and its relation to long-term population data) states that fluctuations in population densities can often be accounted for by external noise. Several empirical models have been suggested to support this view. We followed this by assuming a given population skeleton dynamics (Ricker dynamics and second-order autoregressive dynamics) topped off with noise composed of low- and high frequency components. Our aim was to determine to what extent the modulated population dynamics correlate with the noise signal. High correlations (with time lag -1) were observed with both model categories in the region of stable dynamics, but not in the region of periodic or complex dynamics. This finding is not very sensitive to low-frequency noise. High correlations throughout the entire range of dynamics are only achievable when the impact of the noise is very high. Fitted parameter values of skeleton dynamics modulated with noise are prone to err substantially. This casts doubt as to what degree the underlying dynamics are any more recognizable after being modulated by the external noise. PMID- 11052536 TI - Induced indirect defence in a lycaenid-ant association: the regulation of a resource in a mutualism. AB - Indirect defences involve the protection of a host organism by a mutualistic partner. Threat of predation to the host organism may induce the production of rewards and/or signals that attract the mutualistic partner. In laboratory and field experiments we show that threatened lycaenid butterfly larvae (Plebejus acmon) produce more nectar rewards from their gland and were tended by protective ants twice as much as controls. Ant attendance did not affect the leaf consumption or feeding behaviour of larvae in the absence of predators. Inducible nectar production and indirect defence in this system may be a mechanism by which larvae provide rewards for services when they are needed the most. Such a system may stabilize the mutualistic association between lycaenid larvae and ants by preventing exploitation by either partner. PMID- 11052537 TI - Ant tending influences soldier production in a social aphid. AB - The aphid Pseudoregma sundanica (Van der Goot) (Homoptera: Aphididae) has two defence strategies. It is obligatorily tended by various species of ant and also produces sterile soldiers. We investigated how they allocate their investment in these two strategies. We measured the size, number of soldiers, number and species of tending ant, and number and species of predators in P. sundanica populations. We found that the level of ant tending correlated negatively with soldier investment in P. sundanica. The species of tending ant also influenced soldier investment. We excluded ants from aphid populations and recorded changes in population size and structure over four weeks. Ant exclusion led to population decline and extinction. At the same time, surviving populations showed a significant increase in soldier investment. The data demonstrate that social aphids can adjust their investment in soldiers in direct response to environmental change. PMID- 11052538 TI - Fast and fuel efficient? Optimal use of wind by flying albatrosses. AB - The influence of wind patterns on behaviour and effort of free-ranging male wandering albatrosses (Diomedea exulans) was studied with miniaturized external heart-rate recorders in conjunction with satellite transmitters and activity recorders. Heart rate was used as an instantaneous index of energy expenditure. When cruising with favourable tail or side winds, wandering albatrosses can achieve high flight speeds while expending little more energy than birds resting on land. In contrast, heart rate increases concomitantly with increasing head winds, and flight speeds decrease. Our results show that effort is greatest when albatrosses take off from or land on the water. On a larger scale, we show that in order for birds to have the highest probability of experiencing favourable winds, wandering albatrosses use predictable weather systems to engage in a stereotypical flight pattern of large looping tracks. When heading north, albatrosses fly in anticlockwise loops, and to the south, movements are in a clockwise direction. Thus, the capacity to integrate instantaneous eco physiological measures with records of large-scale flight and wind patterns allows us to understand better the complex interplay between the evolution of morphological, physiological and behavioural adaptations of albatrosses in the windiest place on earth. PMID- 11052539 TI - Mechanical performance of aquatic rowing and flying. AB - Aquatic flight, performed by rowing or flapping fins, wings or limbs, is a primary locomotor mechanism for many animals. We used a computer simulation to compare the mechanical performance of rowing and flapping appendages across a range of speeds. Flapping appendages proved to be more mechanically efficient than rowing appendages at all swimming speeds, suggesting that animals that frequently engage in locomotor behaviours that require energy conservation should employ a flapping stroke. The lower efficiency of rowing appendages across all speeds begs the question of why rowing occurs at all. One answer lies in the ability of rowing fins to generate more thrust than flapping fins during the power stroke. Large forces are necessary for manoeuvring behaviours such as accelerations, turning and braking, which suggests that rowing should be found in slow-swimming animals that frequently manoeuvre. The predictions of the model are supported by observed patterns of behavioural variation among rowing and flapping vertebrates. PMID- 11052540 TI - Hearts, neck posture and metabolic intensity of sauropod dinosaurs. AB - Hypothesized upright neck postures in sauropod dinosaurs require systemic arterial blood pressures reaching 700 mmHg at the heart. Recent data on ventricular wall stress indicate that their left ventricles would have weighed 15 times those of similarly sized whales. Such dimensionally, energetically and mechanically disadvantageous ventricles were highly unlikely in an endothermic sauropod. Accessory hearts or a siphon mechanism, with sub-atmospheric blood pressures in the head, were also not feasible. If the blood flow requirements of sauropods were typical of ectotherms, the left-ventricular blood volume and mass would have been smaller; nevertheless, the heart would have suffered the serious mechanical disadvantage of thick walls. It is doubtful that any large sauropod could have raised its neck vertically and endured high arterial blood pressure, and it certainly could not if it had high metabolic rates characteristic of endotherms. PMID- 11052541 TI - Differential migration and an endocrine response to stress in wintering dark-eyed juncos (Junco hyemalis). AB - The dark-eyed junco (junco hyemalis) exhibits differential migration in autumn that, in general, results in females overwintering south of males, and young within each sex overwintering north of older birds. Individuals overwintering at higher latitudes face less predictable and more challenging environmental conditions. Rapid increases in circulating levels of the energy-regulating glucocorticosteroid, corticosterone, occur in response to environmental stressors. To establish whether the strength of acute corticosterone secretion was correlated with the probability of encountering poor environmental conditions, we compared the corticosterone stress response (e.g. initial plasma concentrations at the time of capture and 30 min later) in dark-eyed juncos overwintering in Mississippi (MS), USA, near the southern limit of their wintering range, with juncos overwintering in New York (NY), USA, near the northern limit of their wintering range. During two winters, 22 males and one female were sampled in NY; 13 males, 12 females and one bird of undetermined sex were sampled in MS. Not unexpectedly, NY birds carried greater fat reserves that resulted in a significantly higher value of energetic condition (mass corrected for wing cord cubed). There was no difference between the two winters sampled at either site, nor was there an effect of sex on patterns of corticosterone secretion in MS birds. With sexes pooled, MS and NY birds had similar baseline corticosterone levels. However, as predicted, NY birds exhibited significantly higher corticosterone concentrations 30 min after capture. These results support the hypothesis that birds wintering in less predictable, more extreme environments show a higher amplitude corticosterone response, which may enable them to adjust their behaviour and physiology more rapidly in response to environmental stressors such as storms. Adrenocortical sensitivity may be a part of the physiological milieu associated with differential migration in juncos; whether it results from endogenous differences in the migratory programmes of individuals or from acclimatization to local environmental conditions remains to be determined. PMID- 11052542 TI - Annual social behaviour of basking sharks associated with coastal front areas. AB - Comparatively little is known about reproductive behaviour in wild sharks as it has proved extremely difficult to study, especially in large pelagic sharks. Here we describe annual courtship-like behaviour in the second-largest fish species, the basking shark (Cetorhinus maximus), from 25 separate episodes observed and tracked during a five-year study period (1995-1999) off south-west England. Social behaviours observed between paired, or three or four, sharks were consistent with courtship behaviours seen in other shark species, namely nose-to tail following, close following, close flank approach, parallel and echelon swimming. Mature individuals between 5 and 8 m total body length (L(T)) exhibited these behaviours whereas smaller sharks (3-4 m L(T)) did not. Lead individuals were identified as female on a number of occasions and interactions were prolonged; the longest continuous observation of socializing was 1.8 h, although intermittent track data indicates bouts may last for up to 5-6 h. Locations of courtship-like behaviour events were not distributed randomly and were significantly associated with thermal fronts. Our results indicate that putative courtship behaviour occurs between May and July along oceanographic fronts, probably as a consequence of individuals aggregating to forage in rich prey patches before initiating courtship. Thus, locating the richest prey patches along fronts may be important for basking sharks to find mates as well as food in the pelagic ecosystem. As courtship-like behaviours occur annually off south-west England we speculate that this region may represent an annual breeding area for this protected species, but mating itself probably takes place at depth as it was not seen at the surface. PMID- 11052543 TI - Carotenoids in the spermatophores of bushcrickets (Orthoptera: Ephippigerinae). AB - During mating male bushcrickets transfer large spermatophores, which have been demonstrated to play an important role in female nutrition and egg production. Until now only relatively unspecific substances such as water and proteins were known to be present within these spermatophores. We found that in the bushcricket Ephippiger zelleri the spermatophores contain substantial amounts of carotenoids (mainly lutein and zeaxanthin) that are also found in the eggs of this species. Carotenoids are well known for their positive effects on survival and reproduction in animals. This is the first example, to our knowledge, where such specific vitamin-like substances were found to be transferred from male to female during mating. PMID- 11052544 TI - Differential effects of word length and visual contrast in the fusiform and lingual gyri during reading. AB - Previous studies have shown differential responses in the fusiform and lingual gyri during reading and suggested that the former is engaged in processing local features of visual stimuli and the latter is engaged in global shape processing. We used positron emission tomography in order to investigate how these regions are modulated by two common variables in reading: word length (three, six and nine letters) and perceptive similarity to the background (high and low contrast). Increasing both word length and visual contrast had a positive monotonic effect on activation in the bilateral fusiform. However, in the lingual gyrus, activation increased with increasing word length but decreased with increasing contrast. On the basis of previous studies, we suggest that (i) increasing word length increases the demands on both local feature and global shape processing, but (ii) increasing visual contrast increases the demands on local feature processing while decreasing the demands on global shape processing. PMID- 11052545 TI - Gating energies and forces of the mammalian hair cell transducer channel and related hair bundle mechanics. AB - We quantified the molecular energies and forces involved in opening and closing of mechanoelectrical transducer channels in hair cells using a novel generally applicable method. It relies on a thermodynamic description of the free energy of an ion channel in terms of its open probability. The molecular gating force per channel as reflected in hair bundle mechanics is shown to equal kT/I(X) x dI(X)/dX, where I is the transducer current and X the deflection of the hair bundle. We applied the method to previously measured I(X) curves in mouse outer hair cells (OHCs) and vestibular hair cells (VHCs). Contrary to current models of transduction, gating of the transducer channel was found to involve only a finite range of free energy (< 10 kT), a consequence of our observation that the channel has a finite minimum open probability of ca. 1% for inhibitory bundle deflections. The maximum gating forces per channel of both cell types were found to be comparable (ca. 300-500 fN). Because of differences in passive restoring forces, gating forces result in very limited mechanical nonlinearity in OHC bundles compared to that in VHC bundles. A kinetic model of channel activation is proposed that accounts for the observed transducer currents and gating forces. It also predicts adaptation-like effects and spontaneous bundle movements ensuing from changes in state energy gaps possibly related to interactions of the channel with calcium ions. PMID- 11052546 TI - Genetic susceptibility to benzene and shortened gestation: evidence of gene environment interaction. AB - This study investigated whether the association between low level benzene exposure and shortened gestation is modified by two susceptibility genes, CYP1A1 and GSTT1. This report includes 542 (302 nonexposed, 240 benzene-exposed) nonsmoking and nondrinking mothers of singleton live births at Beijing Yanshan Petrochemical Corporation between June 1995 and June 1997. Epidemiologic and clinical data and blood samples were obtained from mothers. Multiple linear regression models were used to estimate the associations of benzene exposure and genetic susceptibility with gestational age, adjusting for maternal age, education, parity, stress, passive smoking, prepregnancy weight and height, and infant's sex. Without consideration of genotype, benzene exposure was associated with a decrease in mean gestational age of 0.29 (standard error (SE), 0.12) week. When stratified by the maternal CYP1A1 genotype, the estimated decrease was 0.54 (SE, 0.12) week for the AA group, which was significantly greater (p = 0.003) than that for the Aa/aa group, which showed no decrease in gestational age. When both CYP1A1 and GSTT1 were considered, the greatest decrease was found among exposed mothers with the CYP1A1 AA-GSTT1 absent group (0.79 (SE, 0.25) week) and the CYP1A1 AA-GSTT1 present group (0.50 (SE, 0.22) week). Among the nonexposed, genetic susceptibility alone did not confer a significant adverse effect. This study provides evidence of gene-environment interaction and supports further assessment of the role of genetic susceptibility in the evaluation of reproductive toxins. PMID- 11052547 TI - Invited commentary: on studying the joint effects of candidate genes and exposures. PMID- 11052548 TI - Familial patterns of covariation for cardiovascular risk factors in adults: The Victorian Family Heart Study. AB - The Victorian Family Heart Study was established to address the causes of familial patterns in cardiovascular risk factors. From 1990 to 1996, a representative population sample of 783 adult families (2,959 individuals), each comprising both parents (40-70 years) and at least one natural adult offspring (18-30 years), was recruited in Melbourne, Australia. Included in both generations were 461 monozygotic and dizygotic twins as pairs or singletons. A multivariate normal model was used for pedigree analysis of height, weight, body mass index, diastolic and systolic blood pressure, pulse rate, and total and high density lipoprotein cholesterol. All traits showed evidence for additive genetic variation, explaining from 55% (height) to 26% (pulse) of age- and sex-adjusted variance. An effect persisting into adulthood of shared family environment during cohabitation explained from 39% (body mass index) to 13% (systolic blood pressure) of variance (not nominally significant for diastolic blood pressure). These shared environmental effects were strongest within twin pairs, less so for sibling pairs, and least for parent-offspring pairs (in which an effect was not observed for weight, diastolic and systolic blood pressure, and total cholesterol). On a background of genetic influences, there are periods in early life during which the family environment cements long-term correlations between adult relatives in cardiovascular risk factors. PMID- 11052549 TI - Risk factors for cervical intraepithelial neoplasia in southwestern American Indian women. AB - The authors assessed risk factors for cervical intraepithelial neoplasia (CIN) among southwestern American Indian women using case-control methods. Cases were New Mexico American Indian women with biopsy-proven grade I (n = 190), grade II (n = 70), or grade III (n = 42) cervical lesions diagnosed between November 1994 and October 1997. Controls were American Indian women from the same Indian Health Service clinics with normal cervical epithelium (n = 326). All subjects underwent interviews and laboratory evaluations. Interviews focused on history of sexually transmitted diseases, sexual behavior, and cigarette smoking. Laboratory assays included polymerase chain reaction-based tests for cervical human papillomavirus infection, tests for gonorrhea and chlamydia, wet mounts, and serologic assays for antibodies to Treponema pallidum, herpes simplex virus, and hepatitis B and C viruses. In multiple logistic regression analysis, the strongest risk factors for CIN II/III among American Indian women were human papillomavirus type 16 infection (adjusted odds ratio (OR) = 7.6; 95% confidence interval (CI): 2.4, 23.2), any human papillomavirus infection (OR = 5.8; 95% CI: 3.3, 10.0), low income (OR = 3.3; 95% CI: 1.7, 6.2), and history of any sexually transmitted disease (OR = 2.0; 95% CI: 1.1, 3.5). Unlike previous research, this study found no strong associations between CIN and sexual activity or cigarette smoking. PMID- 11052550 TI - Index for US state tobacco control initial outcomes. AB - Public health tobacco control efforts have increasingly targeted communities in addition to individuals. Before population smoking decreases, effectiveness might be detected from initial outcomes reflecting these efforts, such as higher cigarette prices or more workplace and home smoking restrictions. Presumably, these initial outcomes will eventually influence smoking behavior. State-specific estimates of percentages of the population working or living under smoking bans are available from the 1992-1993 tobacco use supplement to the Current Population Survey, conducted annually by the US Bureau of the Census. In addition, the tobacco industry reports the average state cigarette price yearly. The authors constructed a tobacco control initial outcomes index (IOI) by using values of these variables for each state and correlated it with state-specific adult (aged > or =25 years) and youth (aged 15-24 years) smoking prevalence computed from the Current Population Survey and per capita cigarette consumption data computed from sales and Census Bureau data. Both adult smoking prevalence (r = -0.70) and per capita consumption (r = -0.73) were significantly correlated with the IOI; youth smoking prevalence correlated less well (r = -0.34). Although the analysis is not definitive, deseasonalized 1983-1997 consumption trends for IOI-based tertile groups were divergent beginning in 1993, with the high IOI group showing the greatest decrease. A high relative IOI index may be predictive of future smoking decreases and should be considered when tobacco control efforts are evaluated. PMID- 11052551 TI - One- and two-year predictors of excess weight gain among elementary schoolchildren in multiethnic, low-income, inner-city neighborhoods. AB - Longitudinal studies are needed to increase understanding of the causes of childhood obesity. To identify 1- and 2-year predictors of excess weight gain among preadolescents, the authors conducted a prospective cohort study of fourth- and fifth-grade students in 16 elementary schools located in multiethnic, low income neighborhoods in Montreal, Quebec, Canada, that were participating in the evaluation of a school-based heart health promotion program. Subjects included 2,318 children aged 9-12 years with baseline and 1-year follow-up data and 633 children aged 9-11 years with baseline and 2-year follow-up data. One-year predictors of highest decile of change in body mass index (BMI) identified in logistic regression analyses included baseline BMI of 90th percentile or more (odds ratio (OR) = 2.66, 95% confidence interval: 1.80, 3.94) in boys and baseline BMI of 90th percentile or more (OR = 2.34, 95% confidence interval: 1.46, 3.76), no sports outside school (OR = 1.90, 95% confidence interval: 1.18, 3.06), and playing video games everyday (OR = 2.48, 95% confidence interval: 1.04, 5.92) in girls. Two-year predictors included baseline BMI of 90th percentile or more (OR = 3.26, 95% confidence interval: 1.52, 7.01), no sports outside school (OR = 2.14, 95% confidence interval: 0.96, 4.77), and least active (OR = 2.18, 95% confidence interval: 1.01, 4.71) in boys; only baseline BMI of 90th percentile or more (OR = 2.22, 95% confidence interval: 1.02, 4.81) was significant in girls. Results suggest the need for interventions to promote increased physical activity in children. PMID- 11052552 TI - Dose response of laboratory markers to alcohol consumption in a general population. AB - The dose response to alcohol use of carbohydrate-deficient transferrin (CDT), gamma-glutamyltransferase (GGT), and their combination (gamma-CDT) was studied in an age- and gender-stratified, random sample from Finland in 1997. A linear association with a threshold between alcohol consumption and the three markers was observed. Body mass index was negatively associated with CDT and positively with GGT Age was positively associated with GGT and gamma-CDT In conclusion, CDT appears to be an early phase marker of alcohol consumption. The combined marker, gamma-CDT, was less associated with factors such as body mass index but more strongly correlated with alcohol consumption than were the two markers separately. PMID- 11052553 TI - Urinary 1-methylhistidine is a marker of meat consumption in Black and in White California Seventh-day Adventists. AB - Meat consumption predicts risk of several chronic diseases. The authors validate the accuracy of meat consumption reported by food frequency questionnaires and the mean of eight 24-hour recalls, using urinary methylhistidine excretion, in 55 Black and 71 White Adventist subjects in Los Angeles and San Diego, California, in 1994-1997. 1-Methylhistidine excretion predicts vegetarian status in Black (p = 0.02) and in White (p = 0.005) subjects. Spearman's correlation coefficients between 1-methylhistidine and estimated meat consumption were usually between 0.4 and 0.6 for both food frequency questionnaires and 24-hour recall data. This is despite the chance collection of dietary recalls and urines from omnivores on meatless days. PMID- 11052554 TI - Racial differences in reported Lyme disease incidence. AB - In the United States, the incidence of Lyme disease is considered to be disproportionately high among Whites because of risk of exposure. For assessment of racial differences in Lyme disease incidence and the role of risk exposure, incidence rate ratios (IRRs) for Lyme disease and its manifestations between Whites and African Americans in Maryland and in its focus of endemicity, the Upper Eastern Shore, were calculated. Calculations were based on reported cases of Lyme disease in Maryland during the years 1992-1996. The IRR for Lyme disease between Whites and African Americans was 6.3 (95% confidence interval (CI): 5.0, 8.0), decreasing to 1.8 (95% CI: 1.2, 2.7) for the Upper Eastern Shore. Statewide, there was a significant difference between the White to African American IRR for erythema migrans and for Lyme disease-associated arthritis, at 17.7 (95% CI: 11.2, 27.8) and 2.3 (95% CI: 1.7, 3.2), respectively. On the Upper Eastern Shore, the IRR for arthritis reversed, indicating higher incidence among African Americans than among Whites: IRR = 5.7 (95% CI: 2.4, 13.9) for erythema migrans and IRR = 0.7 (95% CI: 0.4, 1.1) for arthritis. White patients were more likely to have erythema migrans (risk ratio = 2.8, 95% CI: 1.9, 4.1) and less likely to have arthritis than were African Americans (risk ratio = 0.4, 95% CI: 0.3, 0.5). Among all patients, there was a significant negative association between arthritis and erythema migrans. Although much of the racial disparity in incidence rates diminishes in a rural, endemic area, consistent with exposure risk being responsible for much of the variation, a difference remains. This may be due to failure to recognize early disease (erythema migrans) among African Americans, resulting in increased rates of late manifestations. Geographic spread of the disease warrants efforts to increase awareness of Lyme disease and its manifestations among people of color and the health care providers who serve them. PMID- 11052555 TI - Bayesian analysis of an epidemiologic model of Plasmodium falciparum malaria infection in Ndiop, Senegal. AB - Plasmodium falciparum has a complex transmission cycle. Public health planning and research would benefit from the ability of a calibrated model to predict the epidemiologic characteristics of populations living in areas of malaria endemicity. This paper describes the application of Bayesian calibration to a malaria transmission model using longitudinal data gathered from 176 subjects in Ndiop, Senegal, from July 1, 1993, to July 31, 1994. The model was able to adequately predict P. falciparum parasitemia prevalence in the study population. Further insight into the dynamics of malaria in Ndiop was provided. During the dry season, the estimated fraction of nonimmune subjects goes down to 20% and then increases up to 80%. The model-predicted time-weighted average incidences contributed by nonimmune and immune individuals are 0.52 cases per day and 0.47 cases per day, respectively. The median times needed to acquire infection (conversion delay) for nonimmune and immune individuals are estimated at 39 days and 285 days, respectively. PMID- 11052556 TI - Accuracy of alternative approaches to capture-recapture estimates of disease frequency: internal validity analysis of data from five sources. AB - The authors used "internal validity analysis" to evaluate the performance of various capture-recapture methods. Data from studies with five overlapping, incomplete lists generated subgroups whose known sizes were compared with estimates derived from various four-source capture-recapture analyses. In 15 data sets unanalyzed previously (five subgroups of each of three new studies), the authors observed a trend toward mean underestimation of the known population size by 16-25%. (Coverage of the 90% confidence intervals associated with the method found to be optimal was acceptable (13/15), despite the downward bias.) The authors conjectured that (with the obvious exception of geographically disparate lists) most data sets used by epidemiologists tend to have a net positive dependence; that is, cases captured by one source are more likely to be captured by some other available source than are cases selected randomly from the population, and this trend results in a bias toward underestimation. Attempts to ensure that the underlying assumptions of the methods are met, such as minimizing (or adjusting adequately) for the possibility of loss due to death or migration, as was undertaken in one exceptional study, appear likely to improve the behavior of these methods. PMID- 11052557 TI - Impact of pill organizers and blister packs on adherence to pill taking in two vitamin supplementation trials. AB - The impact of pill organizers on pill taking was determined in the Trial of Antioxidant Vitamins C and E (TRACE) Study, a factorial trial of vitamin C and vitamin E supplementation in 184 individuals. Participants were recruited in 1996 1997 and randomized to one of two groups (pill organizer or no organizer) and to one of four supplement groups for 2 months. The pill count (observed/expected X 100%) distribution was similar in the organizer and no organizer group for both vitamins. Mean differences in changes in serum vitamin levels between active and placebo groups did not differ by pill organizer use. The impact of pill organizers and blister packs was compared in another trial, the Vitamins, Teachers, and Longevity (VITAL) Study, in 297 individuals randomized in 1993-1994 to receive study pills either in blister packs or in pill organizers and to take one of two supplements. Among those with lower adherence, the pill count distribution in the blister-pack group exceeded that in the organizer group. Mean differences in serum vitamin E levels between active and placebo groups did not differ by types of pill packaging. In summary, use of blister packs, but not pill organizers, improved adherence as measured by pill counts among those with lower adherence. Neither pill delivery system improved adherence as measured by serum vitamin levels. PMID- 11052558 TI - Self-administered questionnaire compared with a personal diary for assessment of current use of hormone therapy: an analysis of 16,060 women. AB - A personal diary may be more appropriate than a questionnaire for assessing self reported current use of hormone therapy (estrogens, progestagens, or their combination); however, use of a questionnaire is more feasible and less expensive. The authors compared both methods for 16,060 Swedish women aged 45-73 years from the Malmo Diet and Cancer Study (baseline, 1991-1996). In a reliability analysis, the authors investigated the agreement (kappa value) between the questionnaire and the diary regarding current hormone therapy use (yes vs. no), studying the ability to replicate results whether or not they were correct. They also explored associations between discrepancy and individual characteristics. A validity analysis was conducted to determine whether use of the questionnaire achieved an outcome without systematic error (i.e., high specificity and sensitivity); the personal diary was considered the "gold standard." Agreement between both methods was high: 95.5% (kappa = 0.840). The sensitivity was 84.9% and the specificity 97.7%. Higher body mass index and being a widow were associated with agreement, whereas age (50-59 years), use of anxiolytics/hypnotics or opiates, high alcohol consumption, past smoking, and higher educational level were associated with discrepancy. Compared with a personal diary, a simple self-administered questionnaire is a valid method for assessing current use of hormone therapy. PMID- 11052559 TI - A urogynecologic perspective for the 21st century. PMID- 11052560 TI - An evaluation of the efficacy of in-patient bladder retraining. AB - This is a retrospective review of 50 consecutive patients with urinary frequency, urgency and urge incontinence, admitted in 1995 and early 1996 for bladder retraining in the form of timed voiding. At discharge 80% of the women were subjectively cured or satisfactorily improved, but this was significantly reduced to 32% of the 37 who replied to a postal survey between 12 and 29 months (mean interval 21.3 months) later. There were no significant associations between outcome and urodynamic diagnosis, reduced cystometric capacity, length of symptomatology, previous treatment or requirement for additional therapy, but this may have been in part due to small numbers. In conclusion, bladder retraining is a method of treatment for patients with both sensory urgency and detrusor instability which appears to be at least as successful as other modes of treatment for these conditions. PMID- 11052561 TI - Urinary incontinence in women seen in the obstetrics and gynecology clinic. AB - The aim of this study was to present the prevalence of urinary incontinence (UI) in women seen in the obstetrics and gynecology clinic and to describe their behavioral characteristics. A UI-specific questionnaire was administered to 1222 randomly selected women who came to the obstetrics and gynecology clinic of our institution and 15 affiliated hospitals, all located in the island of Kyushu, Japan, for reasons other than incontinence. The prevalence of UI was 31%. Body mass index, parity and menopause significantly influenced the prevalence of UI. The majority of these women (78%) suffered from the stress type of incontinence. A direct relationship was found between the patient's assessment of the severity of her incontinence and the degree of effect on daily activities. The duration, frequency of incontinence and the number of pads used influenced their consultation attitude. Only 25% of these women consulted a physician, often a gynecologist. PMID- 11052562 TI - An assessment of the early surgical outcome and urodynamic effects of the tension free vaginal tape (TVT). AB - Fifty-two women underwent a tension-free vaginal tape (TVT) procedure for genuine stress incontinence (GSI). Preoperative assessment included a detailed medical history, pelvic examination, a 1-hour pad test with a comfortably full bladder, and urinary culture. Thirteen of the 52 women were excluded for various reasons. Both before and 12-24 months postoperatively all patients had a full urodynamic investigation using microtip transducer catheters. The study subjects were instructed to maintain a 1-week baseline urinary diary 1 week before the operation, and postoperative urodynamic assessment as well. The period of follow up ranged from 12 to 24 months (median 19 months). Another 1-hour pad test with a comfortably full bladder and urinary culture were carried out thereafter. Comparisons of the 39 women pre- and postoperatively found a significantly improved 1-hour pad test (34.9 +/- 34.7 vs 8.3 +/- 24.0, P<0.001). Analyzing the urodynamic effects of surgery revealed no significant postoperative changes except for the maximal urethral closure pressure (MUCP) at rest. An objective assessment using a pad test revealed the success rate (cure plus improved) to be 90% (35/39) and the failure rate 10% (4/ 39). TVT can thus be considered a safe and effective procedure for GSI in women. Moreover, the urodynamic effects of surgery were not found to be critical to success. PMID- 11052563 TI - Stress incontinence in women under 50: does urodynamics improve surgical outcome? AB - The aim of the study was to determine whether urodynamic testing improves the outcome of retropubic surgery in women aged 50 or younger. A retrospective study was undertaken of 212 women aged 50 or younger who underwent retropubic surgery at a medical school-affiliated hospital between February 1991 and July 1997. Excluded were patients with prior retropubic urethropexy and known low urethral closure pressures. The surgery was performed by one urogynecologist and two urologists. The minimal evaluation by the urogynecologist was a focused incontinence questionnaire, pelvic neurologic examination, pelvic floor grading, cough stress test, urinalysis, postvoid residual, cotton swab test and supine empty stress test. Full urodynamics consisted of uroflowmetry, subtracted cystometry, urethral closure pressure, cough leak-point pressure and cystourethroscopy. Subjective postoperative follow-up at 14 years was by annual questionnaire. The urogynecologist's patients were in group I (95 women with full urodynamic studies) and group II (36 women with minimal testing). The urologists' patients were in group III (81 women with a very minimal workup and cystourethroscopy). A review of seven variables revealed no difference between the groups. In terms of cured, improved and failed, there was also no difference in outcome. There was a difference in postoperative voiding problems (though not stress incontinence) in group III compared to group I (P= 0.005) and group II (P=0.002). Our conclusion was that all women with stress incontinence should undergo a careful minimal evaluation. In women aged 50 or younger urodynamic studies may be avoided unless there is significant stress incontinence, complex symptoms, a positive supine empty stress test, marked prolapse, or a history of prior retropubic urethropexy. PMID- 11052564 TI - Complete transurethral resection of ulcers in classic interstitial cystitis. AB - Interstitial cystitis (IC) is a chronic disease of obscure etiology. It commonly affects females, who present with symptoms of pain on bladder filling and urinary frequency. There are two types of IC: classic and non-ulcer disease, which differ in many respects, including response to different therapies. In this retrospective study we evaluated the hitherto largest series of patients with classic IC treated by transurethral resection (TUR) of visible ulcers. Altogether 259 TURs of Hunner ulcers were performed on 103 patients: 92 experienced amelioration, and in 40% symptom relief lasted more than 3 years. In the remaining patients, although symptom recurrence was common, the majority responded well to subsequent TUR. In conclusion, TUR has a good outcome in patients with classic interstitial cystitis, and we suggest it as first-line treatment in this patient group. PMID- 11052565 TI - Measurement of postvoid residual urine with portable transabdominal bladder ultrasound scanner and urethral catheterization. AB - The study was a clinical series of 95 ambulatory women with urinary incontinence. After voiding, each subject was scanned with a BladderScan BVI 2500, then catheterized for postvoid residual (PVR) and then scanned again. The mean PVR obtained by ultrasound was 49 ml, significantly larger than the mean PVR of 32 ml obtained by catheterization. Correlation analysis showed that the difference was not related to age, weight, body mass index, parity, pelvic prolapse or prior incontinence surgery, but was associated with prior hysterectomy and uterine prolapse. Regression analysis revealed that the difference was independently related to prior hysterectomy only. Postcatheterization ultrasound detected a mean of 22 ml, suggesting that the difference between the PVR values may be due to residual urine not removed by catheterization. Finally, ultrasound had a sensitivity of 66.7% and a specificity of 96.5% in detecting PVR > or = 100 ml. Portable ultrasound scanners are quick, easy to use, reasonably sensitive, and very specific for determining elevated PVR. PMID- 11052566 TI - Epidemiology and natural history of urinary incontinence. AB - This paper examines or current state of knowledge of the epidemiology of urinary incontinence. The population studied was community-dwelling non-institutionalized persons. The review includes discussion of the prevalence, incidence, natural history and presence of racial and ethnic differences in the epidemiology of urinary incontinence. We also review correlates and potential risk factors that have been revealed in epidemiological studies. Differences between epidemiological and clinical approaches to a health problem, help-seeking behavior and methodological issues for research are also discussed. We have reviewed a large number of completed studies in the field of urinary incontinence, and have emphasized high-quality and population-based studies. We also wished to present studies from a variety of countries. Because of the abundance of studies, only a small fraction can be presented here. Other studies may have equal standards and useful information, but lack of space precludes their inclusion. PMID- 11052567 TI - Transvaginal bladder neck suspension to Cooper's ligament: a review of the literature. AB - The aim of this paper was to review the surgical technique and clinical experience of transvaginal bladder neck suspension to Cooper's ligament. A computerized MEDLINE search identified five English-language articles published between January 1990 and December 1998. The success rate obtained with this procedure is comparable to that obtained with the traditional Burch procedure and ranges between 86.4% and 100%. Postoperative de novo detrusor instability occurs infrequently, with an incidence ranging between 0% and 20%. Complications occur infrequently and postoperative voiding dysfunction is rare. Limited postoperative urodynamic data are available. Transvaginal suspension of the bladder neck to Cooper's ligament combines the technique of a needle suspension with the anatomic effect of the Burch procedure. PMID- 11052568 TI - Vaginal repair of a sigmoidocele. AB - Sigmoidocele is an uncommon accompaniment of pelvic prolapse. It is difficult to detect a sigmoidocele during clinical pelvic examination, and as a consequence a sigmoidocele may be unexpectedly encountered during vaginal repair of pelvic prolapse. The author has discovered and repaired a sigmoidocele during vaginal surgery in 4 patients with either complete procidentia or vaginal vault eversion. The procedure involves a modification of the bilateral sacrospinous vaginal vault fixation using two additional sutures to suspend the sigmoid colon from the sacrospinous ligament. Clinical and functional results have been excellent. This is the first description of a vaginal approach to the repair of a sigmoidocele. PMID- 11052569 TI - Percutaneous bone anchor sling using synthetic mesh associated with urethral overcorrection and erosion. AB - Percutaneous bone anchor bladder neck suspension has been recommended as a less morbid alternative to traditional anti-incontinence procedures. Specifically, it has reported to be associated with shorter duration of hospitalization, catheterization and urinary retention, and equivalent short-term cure rates. Recently, there have been reports of pubic osteomyelitis associated with bone anchor placement, and high incidences of recurrent incontinence. To improve the effectiveness of the procedure the placement of a suburethral synthetic collagen impregnated mesh without tension was recommended. A specific device is included with the kit (Suture Spacer (Microvasive/Boston Scientific Corp., Natick, MA)) to prevent overcorrection of the urethrovesical junction. We present a case of urethral erosion and complete urinary retention secondary to use of a percutaneous bone anchor sling using a ProteGen mesh (Microvasive/Boston Scientific Corp., Natick, MA). Significant postoperative urethral overcorrection was noted despite intraoperative use of the Suture Spacer. PMID- 11052570 TI - Fluid balance therapy of nocturia in women. PMID- 11052571 TI - Reducing gun deaths in the USA. PMID- 11052572 TI - Any room left for disagreement about assisting breech births at term? PMID- 11052573 TI - Risk of asthma in children with exposure to mite and cat allergens. PMID- 11052574 TI - Prevention of acute and recurrent otitis media. PMID- 11052575 TI - Within reach of an end to unnecessary bitterness? PMID- 11052576 TI - From Charles Atlas to Adonis complex--fat is more than a feminist issue. PMID- 11052577 TI - Nomenclature for COX-2 inhibitors. PMID- 11052578 TI - Shuffles. PMID- 11052579 TI - Planned caesarean section versus planned vaginal birth for breech presentation at term: a randomised multicentre trial. Term Breech Trial Collaborative Group. AB - BACKGROUND: For 3-4% of pregnancies, the fetus will be in the breech presentation at term. For most of these women, the approach to delivery is controversial. We did a randomised trial to compare a policy of planned caesarean section with a policy of planned vaginal birth for selected breech-presentation pregnancies. METHODS: At 121 centres in 26 countries, 2088 women with a singleton fetus in a frank or complete breech presentation were randomly assigned planned caesarean section or planned vaginal birth. Women having a vaginal breech delivery had an experienced clinician at the birth. Mothers and infants were followed-up to 6 weeks post partum. The primary outcomes were perinatal mortality, neonatal mortality, or serious neonatal morbidity; and maternal mortality or serious maternal morbidity. Analysis was by intention to treat. FINDINGS: Data were received for 2083 women. Of the 1041 women assigned planned caesarean section, 941 (90.4%) were delivered by caesarean section. Of the 1042 women assigned planned vaginal birth, 591 (56.7%) delivered vaginally. Perinatal mortality, neonatal mortality, or serious neonatal morbidity was significantly lower for the planned caesarean section group than for the planned vaginal birth group (17 of 1039 [1.6%] vs 52 of 1039 [5.0%]; relative risk 0.33 [95% CI 0.19-0.56]; p<0.0001). There were no differences between groups in terms of maternal mortality or serious maternal morbidity (41 of 1041 [3.9%] vs 33 of 1042 [3.2%]; 1.24 [0.79-1.95]; p=0.35). INTERPRETATION: Planned caesarean section is better than planned vaginal birth for the term fetus in the breech presentation; serious maternal complications are similar between the groups. PMID- 11052581 TI - Early exposure to house-dust mite and cat allergens and development of childhood asthma: a cohort study. Multicentre Allergy Study Group. AB - BACKGROUND: In a prospective birth-cohort study, we assessed the relevance of mite and cat allergen exposure for the development of childhood asthma up to age 7 years. METHODS: Of 1314 newborn infants enrolled in five German cities in 1990, follow-up data at age 7 years were available for 939 children. Assessments included repeated measurement of specific IgE to food and inhalant allergens, measurement of indoor allergen exposure at 6 months, 18 months, and 3 years of age, and yearly interviews by a paediatrician. At age 7 years, pulmonary function was tested and bronchial hyper-responsiveness was measured in 645 children. FINDINGS: At age 7, the prevalence of wheezing in the past 12 months was 10.0% (94 of 938), and 6.1% (57 of 939) parents reported a doctor's diagnosis of asthma in their children. Sensitisation to indoor allergens was associated with asthma, wheeze, and increased bronchial responsiveness. However, no relation between early indoor allergen exposure and the prevalence of asthma, wheeze, and bronchial hyper-responsiveness was seen. INTERPRETATION: Our data do not support the hypothesis that exposure to environmental allergens causes asthma in childhood, but rather that the induction of specific IgE responses and the development of childhood asthma are determined by independent factors. PMID- 11052580 TI - Tailored fluorouracil, epirubicin, and cyclophosphamide compared with marrow supported high-dose chemotherapy as adjuvant treatment for high-risk breast cancer: a randomised trial. Scandinavian Breast Group 9401 study. AB - BACKGROUND: Chemotherapy drug distribution varies greatly among individual patients. Therefore, we developed an individualised fluorouracil, epirubicin, cyclophosphamide (FEC) regimen to improve outcomes in patients with high-risk early breast cancer. We then did a randomised trial to compare this individually tailored FEC regimen with conventional adjuvant chemotherapy followed by consolidation with high-dose chemotherapy with stem-cell support. METHODS: 525 women younger than 60 years of age with high-risk primary breast cancer were randomised after surgery to receive nine cycles of tailored FEC to haematological equitoxicity with granulocyte colony-stimulating factor (G-CSF) support (n=251), or three cycles of FEC at standard doses followed by high-dose chemotherapy with cyclophosphamide, thiotepa, and carboplatin (CTCb), and peripheral-blood stem cell or bone-marrow support (n=274). Both groups received locoregional radiation therapy and tamoxifen for 5 years. The primary outcome measure was relapse-free survival, and analysis was by intention to treat. FINDINGS: At a median follow-up of 34.3 months, there were 81 breast-cancer relapses in the tailored FEC group versus 113 in the CTCb group (double triangular method p=0.04). 60 deaths occurred in the tailored FEC group and 82 in the CTCb group (log-rank p=0.12). Patients in the CTCb group experienced more grade 3 or 4 acute toxicity compared with the tailored FEC group (p<0.0001). Two treatment-related deaths (0.7%) occurred in the CTCb group. Six patients in the tailored FEC group developed acute myeloid leukaemia and three developed myelodysplastic syndrome. INTERPRETATION: Tailored FEC with G-CSF support resulted in a significantly improved relapse-free survival and fewer grade 3 and 4 toxicities compared with marrow-supported high-dose chemotherapy with CTCb as adjuvant therapy of women with high-risk primary breast cancer. PMID- 11052582 TI - Treatment of acute otitis media with an antiadhesive oligosaccharide: a randomised, double-blind, placebo-controlled trial. AB - BACKGROUND: Antiadhesive compounds are promising candidates for prevention or treatment of infections. We have investigated the efficacy of such an agent, 3' sialyllacto-N-neotetraose (NE-1530), given intranasally for prophylaxis of acute otitis media and for effect on nasopharyngeal carriage of bacteria. METHODS: We did a randomised, double-blind placebo-controlled study at one study site. 507 healthy children were randomly assigned either NE-1530 (n=254) or placebo (253) as intranasal sprays twice daily during 3 months. The children were examined by the study physicians once a month and during illness. Treatment efficacy was estimated from Cox proportional hazards model. A sample of nasopharyngeal secretion was taken at every visit for culture of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. Adverse events were recorded in study diaries. FINDINGS: At least one event of acute otitis media was diagnosed in 108 (43%) of 254 children in the NE-1530 group and in 86 (34%) of 253 children in the placebo group. The efficacy of treatment was negative, -27% (95% CI -68 to 5; p=0.10). The nasopharyngeal carriage of S pneumoniae, H. influenzae, and M. catarrhalis was not affected by treatment, and the adverse event profiles were almost identical for NE-1530 and placebo. INTERPRETATION: NE 1530 did not have a beneficial effect on the occurrence of acute otitis media or on the nasopharyngeal carriage of bacteria in children. PMID- 11052583 TI - Percutaneous replacement of pulmonary valve in a right-ventricle to pulmonary artery prosthetic conduit with valve dysfunction. AB - BACKGROUND: Valved conduits from the right ventricle to the pulmonary artery are frequently used in paediatric cardiac surgery. However, stenosis and insufficiency of the conduit usually occur in the follow-up and lead to reoperations. Conduit stenting can delay surgical replacement, but it aggravates pulmonary insufficiency. We developed an innovative system for percutaneous stent implantation combined with valve replacement. METHODS: A 12-year-old boy with stenosis and insufficiency of a prosthetic conduit from the right ventricle to the pulmonary artery underwent percutaneous implantation of a bovine jugular valve in the conduit. FINDINGS: Angiography, haemodynamic assessment, and echocardiography after the procedure showed no insufficiency of the implanted valve, and partial relief of the conduit stenosis. There were no complications after 1 month of follow-up, and the patient is presently in good physical condition. INTERPRETATION: We have shown that percutaneous valve replacement in the pulmonary position is possible. With further technical improvements, this new technique might also be used for valve replacement in other cardiac and non cardiac positions. PMID- 11052584 TI - A toxicological surprise. PMID- 11052585 TI - Universal maternal screening for neonatal group B streptococcal disease. AB - Group B streptococcal infection is a leading cause of neonatal morbidity and mortality in the developed world. Data obtained in our region suggest that the incidence in the UK may be higher than previously reported, and together with the results of a pilot study indicate that preventive strategies based on maternal risk factors alone would prevent less than half the cases of neonatal disease. PMID- 11052586 TI - Effect of sleep deprivation on overall 24 h growth-hormone secretion. AB - After sleep deprivation, the blunting of the normal sleep-related growth-hormone (GH) pulse is compensated during the day. Consequently, the amount of GH secreted during a 24 h period is similar whether or not a person has slept during the night. These results argue against the belief that sleep disorders in children can inhibit growth through a daily GH deficit. PMID- 11052587 TI - Effects of transdermal nicotine on cognitive performance in Down's syndrome. AB - Down's syndrome involves age-dependent neuropathological and neurochemical changes similar to Alzheimer's disease, with cholinergic deficits being the most consistent. There is currently no proven treatment for Down's syndrome. We investigated the effect of nicotine-agonistic stimulation with 5 mg transdermal patches, compared with placebo, on cognitive performance in five adults with the disorder. Improvements possibly related to attention and information processing were seen for Down's syndrome patients compared with healthy controls. Our preliminary findings are encouraging, although not generalizable because of small numbers. PMID- 11052588 TI - Coronary heart disease mortality among Arab and Jewish residents of Jerusalem. AB - Information on coronary heart disease (CHD) in the Palestinian population is sparse. We compared mortality rates in the largely Palestinian Arab population of Jerusalem with the Jewish population of the district between 1984 and 1997 based on official Israeli statistics. CHD mortality and all-cause mortality rates were significantly higher among Arab residents than among Jewish residents aged 35-74 years. Whether the excess CHD mortality reflects increased incidence of events, higher case fatality, or both remains to be established. Possible explanations include a higher prevalence of conventional risk factors such as diabetes, obesity, and smoking in Palestinians, stress effects related to the complex political situation and socioeconomic inequalities, and suspected differences in medical care. PMID- 11052589 TI - Sonographic assessment of regional fat in HIV-1-infected people. AB - The routine clinical assessment of lipodystrophy in HIV-1-infected patients is hindered by the absence of easy and reliable methods to measure regional fat. We used sonography to measure subcutaneous fat thickness at three reference skin points (periumbilical, brachial, and malar) and intra-abdominal fat thickness in HIV-1-infected patients with and without lipodystrophy and in healthy controls. Patients without lipodystrophy had less subcutaneous fat than uninfected controls. Sonographic assessment of subcutaneous malar and brachial fat in patients with lipodystrophy was more sensitive and specific than that of intra abdominal fat in the diagnosis of abnormal fat distribution. PMID- 11052590 TI - Improvement of apraxia of eyelid opening by wearing goggles. AB - Apraxia of eyelid opening (ALO) is a non-paralytic motor abnormality characterised by difficulty in initiating the act of eyelid opening without blepharospasm. We found that wearing goggles improved the difficulty of opening eyes in two patients with ALO with parkinsonism. Wearing goggles is a simple method for improving daily life in patients with ALO. PMID- 11052591 TI - Subclinical microtraumatisation of the scrotal contents in extreme mountain biking. AB - Mountain bikers had a high frequency of extratesticular and testicular disorders, which were associated with clinical symptoms in half the bikers. Hence a high rate of repeated microtraumatisation of the scrotal contents must be assumed. PMID- 11052592 TI - Phage therapy--advantages over antibiotics? PMID- 11052593 TI - All change at the top of the national health service. PMID- 11052594 TI - Public hearings on tobacco hailed a success. PMID- 11052595 TI - More deaths from Rift Valley fever in Saudi Arabia and Yemen. PMID- 11052596 TI - Standard of care for alternative medicine. PMID- 11052597 TI - Adverse effects of antiretroviral therapy. AB - Antiretroviral toxicity is an increasingly important issue in the management of HIV-infected patients. With the sustained major declines in opportunistic complications, HIV infection is a more chronic disease, and so more drugs are being used in more patients for longer periods. This review focuses on the pathogenesis, clinical features, and management of the principal toxicities of the 15 licensed antiretroviral drugs, including mitochondrial toxicity, hypersensitivity, and lipodystrophy, as well as more drug-specific adverse effects and special clinical settings. PMID- 11052598 TI - Ireland: breakdown of trust between doctor and patient. AB - By European standards Ireland experiences a very high level of medical litigation. The possible reasons for this are numerous but not yet determined with any certainty. The consequences are quite clear with negative effects on medical practice, on the psychological health of doctors and patients alike, and on finances of the state. There are different patterns of medical litigation throughout Europe. Ireland may be merely anticipating the emerging increase in such actions elsewhere in the region. The present adversarial basis for medical litigation is not wholly without merit and has contributed to professional accountability. Nevertheless, a change in the present system ensuring accountability and fair compensation is required, together with a striving to ensure that trust is rebuilt between doctors and patients. PMID- 11052599 TI - UK: impact of European human rights law. AB - The UK's Human Rights Act 1998, which incorporates into that country's law the European Convention on Rights and Freedoms, came into full operation on Oct 2, 2000. The Act imposes duties on public authorities, who must now justify their position if that is in conflict with a Convention right. Four Articles in the Convention are potential flashpoints in respect of health cases, examples being patients' rights to receive expensive life-saving treatment and disciplinary procedures, including those of the General Medical Council and National Health Service trusts. PMID- 11052600 TI - Clinical picture. Prinzmetal angina. PMID- 11052601 TI - The police medical service of Berlin: doctors or agents of the State? PMID- 11052602 TI - Getting serious about the right to health. PMID- 11052603 TI - Guatemalan government acknowledges human-rights abuses. PMID- 11052604 TI - Self-management of oral anticoagulation. PMID- 11052605 TI - Recombinant DNA vaccines and therapeutics. PMID- 11052606 TI - Strengthening the case for organised trauma care. PMID- 11052607 TI - Strengthening the case for organised trauma care. PMID- 11052608 TI - Cell death in myocardial infarction. PMID- 11052609 TI - Hereditary angioedema and normal C1-inhibitor activity in women. PMID- 11052610 TI - Continuous haemofiltration in acute renal failure. PMID- 11052611 TI - Continuous haemofiltration in acute renal failure. PMID- 11052612 TI - Continuous haemofiltration in acute renal failure. PMID- 11052613 TI - Hypertensive emergencies. PMID- 11052614 TI - Hypertensive emergencies. PMID- 11052615 TI - When trophoblastic disease is suspected. PMID- 11052616 TI - High-dose corticosteroids for asthma. PMID- 11052617 TI - Deferiprone for thalassaemia. PMID- 11052618 TI - Accuracy of references in five leading medical journals. PMID- 11052619 TI - HIV and AIDS, poverty, and causation. PMID- 11052620 TI - Traveller's diarrhoea. PMID- 11052621 TI - Breasts come out of retirement. PMID- 11052622 TI - Decreased cisplatin/DNA adduct formation is associated with cisplatin resistance in human head and neck cancer cell lines. AB - PURPOSE: To evaluate the correlation between cisplatin sensitivity, intracellular glutathione, and platinum/DNA adduct formation (measured by atomic absorption spectroscopy) in a series of seven head and neck cancer cell lines, and to evaluate the effect of biochemical modulation of glutathione on platinum/DNA adduct formation and repair. METHODS: Cisplatin/DNA adducts were measured by atomic absorption spectroscopy. Glutathione content was measured by enzymatic assay and was modulated with buthionine sulfoximine. Apoptosis was measured by double-labeled flow cytometry. RESULTS: Intracellular glutathione concentration was strongly correlated with cisplatin resistance (P = 0.002, R2 = 0.7). There was also a statistically significant inverse correlation between cisplatin/DNA adduct formation and the IC50 for cisplatin in these cell lines. (P = 0.0004, R2 = 0.67). In addition, resistant cells were able to repair approximately 70% of cisplatin/DNA adducts at 24 h, while sensitive cells repaired less than 28% of adducts in the same period. However, despite the positive correlation between cellular glutathione and cisplatin resistance, there was no direct correlation between intracellular glutathione concentration and platinum/DNA adduct formation. Further, depletion of intracellular glutathione by buthionine sulfoximine did not dramatically alter formation of cisplatin/DNA adducts even though it resulted in marked increase in cisplatin cytotoxicity and was associated with increased apoptosis. CONCLUSIONS: These results suggest that glutathione has multiple effects not directly related to formation of cisplatin/DNA adducts, but may also be an important determinant of the cell's ability to repair cisplatin-induced DNA damage and resist apoptosis. PMID- 11052624 TI - Combination of nedaplatin and vindesine for treatment of relapsed or refractory non-small-cell lung cancer. AB - PURPOSE: A phase II study of nedaplatin and vindesine was conducted to evaluate their efficacy and safety for treatment of relapsed or refractory non-small-cell lung cancer (NSCLC). METHODS: Between August 1996 and September 1998, 48 patients who had previously received chemotherapy, thoracic radiotherapy, and/or surgery were enrolled in the study. Patients were required to have an Eastern Cooperative Oncology Group performance status of 0 to 2 and an age between 20 and 79 years. Treatment consisted of nedaplatin (80 mg/m2, day 1) and vindesine (3 mg/m2, days 1 and 8) every 3 to 4 weeks. RESULTS: Of 48 patients, 7 (14.6%) exhibited an objective response. Four (50%) of eight chemotherapy-naive patients had a partial response. However, of the 40 patients who had received prior chemotherapy, a partial response was observed in only 3 (7.5%). At a median follow-up time of 85.1 weeks, the median survival time was 43.6 weeks (95% confidence interval 34.4 52.7) for patients who had received chemotherapy, with a survival rate of 40% at 1 year. Grade 3 or 4 neutropenia occurred in 43 of 48 patients (90%), and neutropenic fever was observed in 3 of the 43 patients, one of whom died of sepsis. Pharmacokinetic and pharmacodynamic analyses of platinum were performed in 43 patients during the first cycle of chemotherapy. Percent reduction in absolute neutrophil count was correlated not only with the area under the plasma ultrafilterable platinum concentration versus time curve (r = 0.41, P = 0.007) but also with the duration of ultrafilterable platinum concentration above 1 microg/ml (r = 0.41, P = 0.007). Patients with progressive disease exhibited a shorter duration of ultrafilterable platinum concentration over 1 microg/ml (P = 0.046) than those with other responses. CONCLUSION: A combination of nedaplatin and vindesine was unsatisfactory as second-line chemotherapy for NSCLC, although the combination was well tolerated. The duration of ultrafilterable platinum concentration above 1 microg/ml was an important pharmacokinetic parameter for predicting both chemotherapy-induced neutropenia and treatment outcome. PMID- 11052623 TI - Camptothecin analogues with enhanced antitumor activity at acidic pH. AB - BACKGROUND: Camptothecin (CPT) is a specific inhibitor of the nuclear enzyme topoisomerase I, which is involved in cellular DNA replication and transcription. Topoisomerase I is therefore an attractive target for anticancer drug development, and two analogues of CPT, topotecan (TPT) and irinotecan (CPT-11), have demonstrated significant antitumor activity in the clinic. This activity is limited, however, by lability of the CPT E ring lactone, which forms the inactive hydroxy acid at physiological pH. The reaction is reversible at acidic pH, which provides a rationale for selectivity, because many solid tumors create an acidic extracellular environment while maintaining a normal intracellular pH. PURPOSE: To exploit the tumor-selective pH gradient to improve the efficacy of CPT-based chemotherapy. METHODS: CPT analogues were evaluated by growth inhibition assay in three human breast cancer cell lines that had been adapted to in vitro culture at acidic pH versus the respective cells cultured at physiological pH. The MCF-7, MDA-MB-231, and MCF-7/hc cell lines represent the hormone-dependent and hormone independent stages of the disease, and a MCF-7 variant that is resistant to the alkylating agent 4-hydroperoxycyclophosphamide (4-HC), respectively. Antiproliferative activity of SN-38 (the active metabolite of CPT-11), and TPT was compared to that of CPT and two CPT analogues, 10,11-methylenedioxy-CPT (MDC), and the alkylating derivative, 7-chloromethyl-10,11-MDC (CMMDC). RESULTS: In general, MDC was the most potent and TPT or CPT the least potent analogue, regardless of pH. However, if the comparison was based on magnitude of potentiation by pH, a different rank order emerged. CPT was modulated 4-fold; MDC, SN-38, and TPT were each modulated 5- to 6-fold, while the activity of CMMDC was increased 10- to 11-fold by acidic pH in MCF-7 lines, and 65-fold in MDA-MB 231 cells. Thus MDC was the superior CPT analogue based on potency, but CMMDC was the best candidate for pH modulation. Drug specificity was also observed. While the alkylating agent, 4-HC, was 2- to 3-fold more active at acidic pH, modulation was not observed for 5-fluorouracil, doxorubicin, or paclitaxel. Preliminary mechanism studies indicated that pH modulation of CPT analogues was directly correlated to intracellular levels of glutathione. In addition, protein associated DNA strand breaks were more rapidly induced at acidic pH. CONCLUSION: These results suggest that CPT-based drug development and resulting chemotherapy could benefit from evaluation of differential activity at acidic versus physiological pH. Analogues have been identified that could have improved therapeutic indices based on the pH gradient that selectively exists in human tumors. PMID- 11052625 TI - Liposomal daunorubicin plasmatic and renal disposition in patients with acute leukemia. AB - Liposomal formulations of anthracyclines have been developed to increase their delivery to solid tumors while reducing toxicity in normal tissues. DaunoXome (DNX, NeXstar) is a liposomal-encapsulated preparation of daunorubicin registered for treatment of Kaposi's sarcoma that during prior in vitro studies showed a toxicity to leukemic cells at least comparable to that of free daunorubicin. The aim of our study was to determine DNX pharmacokinetics in 11 poor-risk patients with acute leukemia treated with DNX 60 mg/m2 IV on days 1, 3, and 5. Blood and urine samples were collected at appropriate intervals after each of the three DNX administrations. The total amount of daunorubicin (free and entrapped) (t-DNR) and of its metabolite daunorubicinol (DNRol) was assayed by HPLC. The main pharmacokinetic parameters (t1/2alpha 4.54 +/- 0.87 h; VdSS 2.88 +/- 0.93 l/m2; Cl 0.47 +/- 0.26 l/h/m2) showed that in patients with acute leukemia liposomal entrapped daunorubicin pharmacokinetics greatly differed from that observed for the conventional formulation. In fact, DNX produced mean plasma AUC levels (t-DNR AUC0-infinity 456.27 +/- 182.64 microg/ml/h) about 100- to 200-fold greater than those reported for the free drug at comparable doses due to a very much lower total body clearance. Volume of distribution at steady state was 200-to 500-fold lower than for the free drug. Plasma AUC of DNRol (17.62 +/- 7.13 microg/ml x h) was similar to or even greater than that observed with free daunorubicin for comparable doses. Cumulative urinary excretion showed that about 6% and 12% of the total dose of DNX administered was excreted in urine as daunorubicin and daunorubicinol, respectively. No major toxicity was encountered. Therefore, pharmacokinetic characteristics suggest that DNX may be more convenient than free daunorubicin in the treatment of acute leukemia. In fact, liposomal formulation may allow a reduction of daunorubicin captation in normal tissues. thus minimizing toxicity at least for the parent drug, and guarantee an unimpeded access to leukemic cells in the bloodstream and bone marrow, thus theoretically improving efficacy. PMID- 11052626 TI - Selective modulation of P-glycoprotein's ATPase and anion efflux regulation activities with PKC alpha and PKC epsilon in Sf9 cells. AB - The modulation of P-glycoprotein's (Pgp) ATPase activity and its ability to regulate swelling-activated 125I efflux, by PKC alpha and PKC epsilon, was examined in insect cells. Recombinant baculovirus was used to express human Pgp in Sf9 cells and Pgp was also coexpressed with either PKC alpha or PKC epsilon. ATPase assays showed the enzyme activity of Pgp to be elevated during co expression with the Ca2+ dependent isoform PKC alpha, but not with the Ca2+ independent variant PKC epsilon. Furthermore, neither isoform, when co-expressed with Pgp, altered the swelling-activated efflux of 125I from Sf9 cells. However, in cells co-expressing Pgp/PKC (alpha or epsilon), pre-treatment with the phorbol ester TPA significantly reduced the swelling-activated 125I efflux with both PKC isoforms. Our results suggest that phosphorylation with the Ca2+ independent variant PKC epsilon does not regulate the ATPase activity of Pgp and that stimulation of PKC with TPA alters the swelling-activated efflux of anions from insect cells expressing Pgp. PMID- 11052627 TI - Expression of pRB, cyclin/cyclin-dependent kinases and E2F1/DP-1 in human tumor lines in cell culture and in xenograft tissues and response to cell cycle agents. AB - PURPOSE: Cell cycle regulatory components are interesting targets for cancer therapy. Expression of pRb, cyclin D1, cdk4, cyclin E, cdk2, E2F1 and DP-1 was determined in MCF-7 and MDA-MB-468 breast carcinoma cells, H460 and Calu-6 non small cell lung carcinoma cells, H82 and SW2 small cell lung carcinoma cells, HCT116 and HT29 colon carcinoma cells and LNCaP and DU-145 prostate carcinoma cells. METHODS: For Western blotting, the ratio with actin expression was used to normalize the data; all lines were run on the same gels. RESULTS: In cell culture, pRb was not detected in MB-468 and H82 was low in SW2 and DU-145 and highest in HCT116; in tumors, pRb was not detected in MB-468, H82, SW2, and DU 145 and was highest in LNCaP and Calu-6. Cyclin D1 was not detected in SW2 cells in culture, was low in MB-468 and H82, and was highest in LNCaP and H460; in tumors, cyclin D1 was low in MB-468, H460, SW2 and DU 145, and was highest in LNCaP. In cell culture, cdk4 was lowest in Calu-6, HCT116, HT29 and DU-145 and highest in H82 and SW2; in tumors, cdk4 was low in MCF-7, MB-468, H460, Calu-6 and HCT116 and was very high in the SW2. Expression of cyclin E was very low in MCF-7 and HT29 and high in H460 in culture and was very low in MCF-7, H460, Calu 6, H82, HT29 and DU-145 in tumors and high in HCT116 and LNCaP. In cell culture, E2F1 was lowest in MB-468, Calu-6, HT29 and DU-145 cells and highest in LNCaP cells; in tumors, E2F1 was lowest in MCF-7, MB-468 and Calu-6 and highest in LNCaP. In cell culture, DP-1 was lowest in MB-468, HCT116 and HT29 and highest in SW2. The MCF-7 and MB-468 lines were most resistant to flavopiridol and olmoucine and the H460 and Calu-6 lines were most resistant to genistein. The SW2 tumor was most responsive to flavopiridol and olomoucine. CONCLUSIONS: There is a high degree of variability in the expression of cell cycle components in human tumor cell lines, resulting in complexity in predicting response to cell cycle directed agents. PMID- 11052628 TI - In vitro evaluation of newly developed chalcone analogues in human cancer cells. AB - PURPOSE: Among flavonoids, chalcones have been identified as interesting compounds having chemopreventive and antitumor properties. We studied a panel of newly developed chalcone analogues (S1-S10) using MDA-MB 231 and MCF-7 ADRr breast cancer cells and the T-leukemic Jurkat cell line. Quercetin was used as the reference compound. METHODS: Antiproliferative activity was evaluated by cell counts performed after 72 h of exposure to the drugs. DNA analysis and redox activity were evaluated using flow cytometry. Apoptosis was assessed by morphological analysis, using YOYO-1 as DNA dye; p-glycoprotein function was ascertained by quantitating the efflux of rhodamine 123. RESULTS: All cells were sensitive to chalcone analogues yielding IC50 in micromolar concentrations with the following order regardless of the multidrug resistance (MDR) status: S1 > S2 > quercetin. S1 and S2, the most active compounds, were selected to evaluate their effect on the cell cycle, apoptosis, redox activity, and modulation of the p-glycoprotein function. No significant perturbation in cell cycle was seen with concentration up to 1 microM after 24 h. After 72 h a slight increase in G2/M block and DNA fragmentation occurred at 10 microM. Morphological analysis of apoptosis showed that chalcone analogues induced apoptosis to a higher extent than quercetin. Redox analysis demonstrated that all substances were able to increase intracellular thiol levels, which returned to baseline value after 24 h for all drugs except quercetin. Production of reactive oxygen species was essentially unaffected by all compounds. Finally, in MDR-positive MCF-7 ADRr cells chalcone analogues were unable to modulate p-glycoprotein function while quercetin was able to. CONCLUSIONS: Newly developed S1 and S2 chalcones have a different but higher antitumor activity than quercetin and could be considered as potential new anticancer drugs. PMID- 11052629 TI - Inhaled aerosolization of all-trans-retinoic acid for targeted pulmonary delivery. AB - Retinoids have shown promising activity for both cancer chemoprevention and as a treatment for emphysema. However, chronic oral administration of these drugs is limited by systemic side effects, including hepatic dysfunction, skeletal malformations, hyperlipidemia. hypercalcemia, and other reactions. In order to improve the pulmonary targeting of this potentially useful therapy, we developed a system for aerosolization of retinoids that substantially increased their local bioavailability. We compared the biodistribution and pharmacokinetics of an inhaled formulation of all-trans-retinoic acid (all-trans-RA), which was packaged in a metered dose inhaler, following both intratracheal (IT) and intravenous (IV) administration in male Sprague-Dawley rats. After drug administration, anesthetized animals were killed at 5 min, and at 1, 2, 4, 6 and 24 h. Plasma and emulsified samples of liver and lung tissues were dissected, extracted, and frozen prior to measurement of all-trans-RA concentration by high-performance liquid chromatography (HPLC). Aerosolization and IT injection of all-trans-RA resulted in a significantly longer pulmonary half-life of the drug (both 5-17 h), lower peak serum concentrations (aerosol 71 +/- 31 ng/ml, IT 68 +/- 50 ng/ml), and lower liver levels (aerosol 111 +/- 28 ng/g, IT 753 +/- 350 ng/g) than the same dose administered IV (2 h, 838 +/- 56 ng/ml, 4,258 +/- 1,006 ng/g, respectively; P < 0.05 for each comparison). Histologic examination of lungs and trachea showed no focal irritation attributable to the drug after single-dose administration. These results suggest that aerosolization of retinoids may offer a practical alternative to systemic oral administration for chemoprevention trials or treatment of lung diseases. This method may substantially increase the therapeutic index of these compounds by reducing systemic complications associated with long-term dosing. PMID- 11052631 TI - Multiple factors other than p53 influence colon cancer sensitivity to paclitaxel. AB - PURPOSE: To determine factors which influence the sensitivity of human colorectal carcinoma cell lines to paclitaxel. METHODS: The paclitaxel sensitivity of ten human colorectal carcinoma cell lines, and a panel of RKO colon carcinoma cell lines, isogenic except for p53 status, were studied. The inhibitory concentrations causing a 50% decrease in growth (IC50) were assayed after 3, 24, and 96 h after paclitaxel exposure. The doubling time (DT) and cell cycle parameters of cells were also measured. The expression of the multidrug resistance glycoprotein-1 (MDR-1), bcl-2 and bax was quantitatively assessed by immunoblotting. RESULTS: Mean IC50 values at 24 and 96 h drug exposure were about 1.5 logs lower than the IC50 values at 3 h, regardless of the p53 status. No difference was found between the IC50 values of wild-type and mutant p53 cells, or among the RKO panel of cells. Correlation analysis showed that: (1) resistance was associated with longer DTs, but this was generally abated by a 96-h exposure; (2) with a 3-h exposure, the combination of MDR, bcl-2 and bax parameters with DT (DT + MDR + bcl-2 bax) best correlated with IC50 values (r = 0.77); (3) with a 96 h exposure, in spite of the generally decreased IC50 values, a combination of MDR 1, bcl-2 and bax parameters (MDR + bcl-2-bax) best correlated with the IC50 values (r = 0.71). CONCLUSIONS: These results suggest that the exposure duration, DT, and expression of MDR-1, bcl-2 and bax each contribute to paclitaxel sensitivity of human colorectal carcinoma cells. In assessing paclitaxel drug resistance, multiple factors should always be considered. There may be a therapeutic window for taxanes in colon cancer by optimizing pharmacokinetics and modulating MDR-1 and bcl-2 resistance factors. PMID- 11052630 TI - A phase I clinical and pharmacokinetic study of the dolastatin analogue cemadotin administered as a 5-day continuous intravenous infusion. AB - PURPOSE: The dolastatins are a class of naturally occurring cytotoxic peptides which function by inhibiting microtubule assembly and tubulin polymerization. Cemadotin is a synthetic analogue of dolastatin 15 with potent antiproliferative and preclinical antitumor activity. This report describes a phase I study to evaluate the administration of cemadotin to adult cancer patients by a 5-day continuous intravenous (CIV) infusion. METHODS: All patients had histologically confirmed refractory solid tumors. The dose was escalated from an initial level of 2.5 mg/m2 (0.5 mg/m2 daily) according to a modified Fibonacci algorithm. A minimum of three patients was evaluated at each dose level until the maximum tolerated dose (MTD) was established. Treatment was repeated every 21 days until patients were removed from the study due to toxicity or disease progression. Drug related toxicities were evaluated and graded by the U.S. National Cancer Institute's Common Toxicity Criteria. A radioimmunoassay (RIA) that detected both the parent drug and its metabolites with an intact N-terminal region of the molecule was used for pharmacokinetic studies. RESULTS: Twenty heavily pretreated patients received a total of 40 courses of cemadotin over five dose levels ranging from 2.5 to 17.5 mg/m2. Reversible dose-related neutropenia was the principal dose-limiting toxicity and 12.5 mg/m2 was established as the MTD. Nonhematologic toxicities attributed to the drug were moderate, and there was no evidence of the cardiovascular toxicity noted in the prior phase I studies of cemadotin given IV as a 5-min injection or 24-h infusion. There were no objective antitumor responses. Time courses of the cemadotin RIA equivalent concentration in whole blood were defined in 14 patients during the first cycle of therapy. The RIA-detectable species exhibited apparent first-order pharmacokinetics across the entire range of doses. The mean +/- SD of the observed steady-state blood concentration at the 12.5 mg/m2 MTD was 282 +/- 7 nM (n = 3). Blood levels decayed monoexponentially following the end of the infusion, with a mean half life of 13.2 +/- 4.3 h (n = 14) in all patients. Mean values (n = 14) of the total blood clearance and apparent volume of distribution at steady state were 0.52 +/- 0.09 lh/m2 and 9.9 +/- 3.3 l/m2, respectively. CONCLUSIONS: The cardiotoxic effects of cemadotin were completely avoided by administering it as a 120-h CIV infusion. Thus. cardiovascular toxicity appears to be associated with the magnitude of the peak blood levels of the parent drug or its metabolites, whereas myelotoxicity is related to the duration of time that blood levels exceed a threshold concentration. Nevertheless, the data acquired during the extensive clinical experience with cemadotin requires careful examination to assess whether advancing this compound into disease-oriented efficacy studies is merited. PMID- 11052632 TI - A phase I study of prolonged ambulatory infusion carboplatin with oral etoposide showing activity in prostate cancer. AB - PURPOSE: This phase I study aimed to establish the dose for phase II trials of a dose-intense outpatient regimen of ambulatory carboplatin and oral etoposide. PATIENTS AND METHODS: Cohorts of three patients received escalating doses of carboplatin 15, 20, and 23 mg/m2/day as a 3-week continuous ambulatory infusion with oral etoposide initially at 50 mg/day. Patients entered had prostate, colon, head and neck, breast, unknown primary cancers and mesothelioma. RESULTS: At 23 mg/m2 of carboplatin, two patients had WHO grade 3 lethargy and myelosuppression, which were the dose-limiting toxicities. Six patients were entered at the dose recommended for phase II studies, carboplatin 20 mg/ m2/day and etoposide 50 mg/day for 21 days repeated every 6 weeks. This was well tolerated except for one patient with multiple bone metastases from prostate cancer experiencing grade 4 myelosuppression and a single patient with grade 3 constipation. Seven patients with hormone-resistant prostate cancer were entered into the study, one at 15 mg/m2, four at 20 mg/m2 and two at 23 mg/m2 of carboplatin, and received a median of four cycles of treatment. The only responses were seen in prostate cancer where there were two partial responses in patients with soft tissue predominant disease. Five patients who could be evaluated with initially elevated PSA exhibited falls of > or 50% after receiving the chemotherapy. All but one patient with prostate cancer experienced significant reduction in pain levels. The median time to progression of the patients with prostate cancer was 4 months. CONCLUSIONS: Ambulatory infusion carboplatin and oral etoposide is a tolerable dose-intense outpatient regimen which warrants further testing in phase II trials including hormone-resistant prostate cancer. PMID- 11052633 TI - Audiovisual sexual stimulation by virtual glasses is effective in inducing complete cavernosal smooth muscle relaxation: a pharmacocavernosometric study. AB - Audiovisual sexual stimulation (AVSS) is frequently employed to promote cavernosal smooth muscle relaxation (SMR) in hemodynamic diagnostic settings for erectile dysfunction. Our aim has been to adapt conventional AVSS to the particular test conditions of pharmacocavernosometry and pharmacocavernosography (DICC), by the use of virtual glasses. Thirty-seven consecutive patients undergoing DICC were randomized in two groups: no-AVSS and AVSS through commercially available virtual glasses (VG-AVSS) with tri-dimensional capabilities and stereophonic headphones. Such device partially excludes the patient from the surrounding environment. In both groups a standard dose of vasoactive agents was intracavernosally administered, and possibly repeated (re dosing), until complete SMR was obtained (3 doses/patient maximum). Psychometric tests (State Trait Anxiety Inventory and ad hoc visual analogue scales for embarrassment, stress and pain) were administered before and after DICC. The no AVSS group consisted of 18 patients, the AVSS group of 19. Number of needed vasoactive agent doses: in the no-AVSS group 6 patients needed 1 dose, 3 patients 2, 9 patients 3 (mean dose number: 2.17); in the AVSS group 15 patients needed 1 dose, 1 patient 2, 3 patients 3 (mean dose number: 1.37). The difference in the number of doses used in the two groups was statistically significant (Student's t test P = 0.007). Complete SMR, regardless of the number of used doses: in the no AVSS group 9 patients (50%) achieved complete SMR, in the AVSS group 16 patients (84.2%). The difference in the two groups was statistically significant (chi square P = 0.026). From evaluated psychometric measures no statistically significant difference between the two groups was detected. VG-AVSS significantly promotes complete SMR without increasing test related stress or anxiety. Its induced arousal suggests the possibility of performing dynamic evaluations of the erectile function with the oral agent sildenafil in place of intracavernosally administered vasoactive agents. VG-AVSS furthermore constitutes a promising tool for the investigation of normal physiology and pathophysiology of female sexual function. PMID- 11052634 TI - The conditioned response erection (CRE)--a new approach to modelling erectile responses in the rat. AB - Several animal models are currently used in erectile (dys)function research; these models fail to account for the conditions involving the more spontaneous erections in humans. Recently, we observed an increase in the number of 'spontaneously' occurring erections in rats with previous exposure to apomorphine (APO), a centrally acting drug that initiates penile erections and yawns. Based on this observation, we designed a series of experiments to characterize the development of enhanced, non-apomorphine-induced erections or 'spontaneous' erectile responses to vehicle administration in rats with previous exposure to APO. We further examined the effects of castration on these conditioned erections. Naive (ie never received APO) rats were administered vehicle (1 ml/kg saline) to determine the frequency of baseline erections and yawns. An alternating series of APO (80 microg/kg s.c.) and vehicle administrations were performed over several days and subsequent erectile and yawning responses were recorded. Following 3 sets of 3 APO administrations (with vehicle administered between sets), and the 3rd vehicle administration, these rats were then surgically castrated and allowed 30 days to recover. Following this, APO was administered 3 times to determine erectile and yawning responses post-castration, followed by vehicle administration to determine the effects of castration on conditioned APO responses. The major findings were: (1) that although naive rats had a basal spontaneous erectile response (0.75 +/- 0.88; 4 of 8 rats with at least one erection), repetitive administration (up to 22 treatments) of the central initiator apomorphine significantly increased the number of erections (1.8 +/- 0.7; 7 of 8 rats with at least one erection) and yawning (2.5 +/- 2.47) responses to vehicle administration; and (2) both spontaneous yawning and erectile responses were found to be androgen dependent since castration dramatically lowered the number of erections (0.13 +/- 0.35; 1 of 8 rats with at least one erection) and yawns (0). Therefore, this method of producing erections without a pharmacological manipulation provides an additional animal model which can be used in conjunction with the APO-induced erections in characterizing the physiology and pathophysiology of erectile function in conscious rats. PMID- 11052635 TI - Multicenter study of the treatment of erectile dysfunction with transurethral alprostadil (MUSE) in Korea. AB - A Korean multicenter study was conducted to assess the effectiveness of transurethral alprostadil with MUSE in 334 subjects with chronic erectile dysfunction (ED) who were enrolled in 21 clinical centers. Patients with psychogenic impotence comprised about 30% of subjects. Intraurethral alprostadil was titrated in a stepwise fashion in the clinics from 250 to 500 or 1000 mcg based on erectile response and tolerability. The erectile responses were evaluated using an erection assessment scale (score of 1-5). The dose that produced a maximal penile response of score 5 (full rigid erection) or 4 (full tumescence, partial rigidity) was selected for home treatment. Patients who showed partial erection (score of 3) with 1000 mcg were also included in the home treatment group. In-clinic phase: 198 men (59.3%) had maximal penile responses of score 4 or 5. The rate of maximal responses was not related to patient age, etiology or duration of the ED. A total of 228 (68.3%) men progressed to home treatment. The overall level of comfort of the transurethral alprostadil was rated as uncomfortable or very uncomfortable in 12%. Home phase: During the two month period of home treatment, 178 (78.1%) men had successful sexual intercourse at least once, and 78.2% of administrations (1976) resulted in successful intercourse. The main causes of drop-out were insufficient erectile response in 27 men (11.8%), adverse reactions (mostly penile or urethral pain) in 7 (3.1%) or both in 7 (3.1%). In conclusion, transurethral alprostadil could be a suitable treatment option for patients with ED regardless of age and etiology of ED. Efficacy in an Asian population (Korea) is comparable to that reported previously in Caucasians. PMID- 11052636 TI - The value of testing pudendal nerve conduction in evaluating erectile dysfunction in diabetics. AB - We conducted an open prospective study on the value of testing pudendal nerve conduction (PNC) in 45 diabetic and 32 nondiabetic men with documented erectile dysfunction (ED) of at least six months duration. All subjects underwent PNC by the same investigator using the Medcelec/TECA Sapphire device with calibration parameters of sweep 10 ms/div an amplitude of 200 microV/div. No statistically significant differences was found in the mean bulbocavernosus reflex (BCR) latencies between the nondiabetics (33.6 ms/div +/- 4.1) and the diabetics (37.5 ms/div +/- 9.1) (P > 0.05). Our results show that electrophysiological measurement of the BCR in diabetics is not a useful diagnostic test and emphasize the importance of a multifactorial evaluation of diabetic ED. PMID- 11052637 TI - Misoprostol induces relaxation of human corpus cavernosum smooth muscle: comparison to prostaglandin E1. AB - Prostaglandin E1 (PGE1) relaxes trabecular smooth muscle by interacting with specific G-protein coupled receptors on human corpus cavernosum smooth muscle and increasing intracellular synthesis of cAMP. Misoprostol (Cytotec), is an oral prostaglandin E analogue. The purpose of this study was to compare the functional activity of misoprostol with PGE1 in human corpus cavernosum and cultured human corpus cavernosum smooth muscle cells. Misoprostol, misoprostol free acid or PGE1 induced dose-dependent relaxations in strips of human corpus cavernosum. At concentrations greater than 10(-6) M, tissue recontraction was observed with all three agents. This was abrogated by pretreatment with the thromboxane A2 receptor antagonist SQ29,548. From these observations, we conclude that misoprostol is activated by human corpus cavernosum in situ and relaxes phenylephrine precontrated tissue strips in vitro. This relaxation response is mediated by the increased cAMP synthesis by these agents. PMID- 11052639 TI - Horizontal plication after vertical tunical incisions for the correction of congenital penile curvature. AB - The objective of this retrospective study is to evaluate the surgical outcome of correction of congenital penile curvature, via multiple vertical incisions in the tunica albuginea that are sutured horizontally using simple inverted 2-0 PDS sutures. The study included 22 men with congenital penile curvature. The surgeries were performed in three general hospitals. The procedures straightened the penile shaft in all cases but a degree of curvature recurred in three cases. No operative or postoperative complications occurred and no reoperations were needed. Four patients complained of penile shortening. We conclude that horizontal plication after vertical corporal incisions is safe and effective in the treatment for congenital penile curvature without hypospadius. PMID- 11052638 TI - Atherosclerosis-induced chronic arterial insufficiency causes clitoral cavernosal fibrosis in the rabbit. AB - In our previous studies we found that aging-associated fibrosis of clitoral cavernosal tissue correlated with the prevalence of cardiovascular disease in elderly women. The aim of this study was to determine specifically, arterial insufficiency-related structural changes of clitoral cavernosal tissue in a rabbit model. New Zealand white female rabbits were divided into clitoral cavernosal ischemia (CCI, n = 5) and control (n = 5) groups. The CCI group underwent balloon endothelial injury of the iliac arteries and received 0.5% cholesterol diet. The control group received a regular diet. After 16 weeks, arteriography was performed then the animals were sacrificed. The iliac arteries and the entire clitoris were removed. Cross-sections of the iliac arteries and clitoris were processed for histologic evaluation The percentage of smooth muscle and connective tissue in trichrome stained sections of clitoral cavernosal tissue was determined by computer-assisted histomorphometry. Arteriography revealed diffused occlusive disease in the common iliac, internal iliac and pudendal arteries in the CCI group. Histology showed that arterial occlusive disease spreads from the site of balloon injury to the smaller branches involving the clitoral cavernosal arteries. Diffuse fibrosis was observed in the clitoral cross sections of the CCI group. The percentage of clitoral cavernosal smooth muscle (mean +/- standard error) in the CCI group (53% +/- 0.9%) was significantly decreased compared with the control group (62% +/- 0.8%) (P = 0.0001). Chronic clitoral cavernosal ischemia causes significant fibrosis and loss of smooth muscle in the clitoral cavernosal tissue. These findings suggest that chronic clitoral cavernosal arterial insufficiency may play a role in the pathophysiology of female sexual arousal disorders. PMID- 11052640 TI - Kallmann's syndrome: clues to clinical diagnosis. AB - Hypogonadotropic patients may visit pediatricians, general practitioners, endocrinologists or urologists, presenting with microphallus, cryptochidism or pubertas tarda and delayed bone maturation. Congenital hypogonadotropic hypogonadism is characterized, apart from small testes, by the constellation of low serum levels of testosterone, LH and FSH. Kallman's syndrome is characterized by congenital hypogonadotropic hypogonadism with midline defects such as anosmia (a deficiency of the sense of smell). The first case report dates back to 1856, and genetic defects causing the syndrome have been recently described. The diagnosis can be clinically suspected and is established by confirming hormonal studies. PMID- 11052641 TI - Penile fracture repair: assessment of early results and complications using color Doppler ultrasound. AB - The aim of this study was to determine early results and complications of penile fracture treated with immediate surgical repair by means of color Doppler ultrasound study. Four patients with the clinical features of penile fracture were submitted to immediate surgical exploration via a subcoronal incision with repair of the torn cavernosal albuginea (unilateral in three cases, bilateral in one case) and anastomosis of the transected urethra (one case). Color Doppler ultrasound (CDUS) was performed by means of an Acuson 128XP/10 using a 7-10 MHz extended frequency linear array transducer. Erectile function at five months follow-up was reported as normal by two patients (age 59 and 55 y), slightly decreased in one case (bilateral partial cavernous fracture + total urethral transection in a 32 y old) and weak in one case (51 y old). In the latter two, the investigation included a dynamic phase following a 10 mcg PGE injection. B mode ultrasound showed no fibrotic changes in relation to the long-term absorbable suture material. Baseline CDUS demonstrated full length integrity of the cavernous arteries in all patients. The CDUS dynamic study was entirely normal in the patient with weak erection while showed a continuous venous leak in the patient with bilateral cavernosal rupture and transected urethra. We conclude that despite the onset of erectile failure in two out of four patients, there was no evidence of arteriogenic impotence in any patients with major penile fracture and thus we advocate early simple repair without any microsurgical exploration of the cavernosal arteries. PMID- 11052642 TI - Intracavernosal therapy for erectile failure, impact of treatment and reasons for drop-out and dissatisfaction. PMID- 11052643 TI - Vasculogenic impotence and cavernosal oxygen tension. PMID- 11052644 TI - Stimulation of collagen production in an in vitro model for Peyronie's disease. PMID- 11052645 TI - Re-dosing of prostaglandin-E1 versus prostaglandin-E1 plus phentolamine in male erectile dysfunction: a dynamic color power Doppler study. PMID- 11052646 TI - Immune aspects of Bartonella. AB - Bartonella species have been recognized as important human pathogens only recently. Until the early 1990s, this genus was represented by one species, Bartonella bacilliformis. The recent identification of other Bartonella species as the agents of cat-scratch disease and bacillary angiomatosis has left little doubt of their emerging importance as opportunistic human pathogens. Over the last decade, extensive research has been performed on Bartonella species, resulting in an explosion in our knowledge of the genetic diversity of this genus. Unusual aspects of disease sequelae have fueled worldwide interest in defining the natural history, pathology, and molecular biology of Bartonella species. While much information about these interests has been presented, the advancement of immunological knowledge regarding Bartonella species has been slow. This review discusses immunological data on Bartonella species, focusing on the three primary human pathogens of this genus: B. bacilliformis, B. quintana, and B. henselae. PMID- 11052647 TI - Cork taint in wine: scientific knowledge and public perception: a critical review. AB - The manufacturing process of cork stoppers includes a stabilization period of the cork slabs, following boiling, during which mold growth completely covers the cork slabs. This process has been used traditionally for several decades; however, due to the possibility of certain molds isolated from cork to produce off flavor compounds, especially 2,4,6-trichloroanisole and 2,3,4,6 tetrachloroanisole, recently cork stoppers are being unsoundly targeted with the accusation of inducing cork taint in wine. This article reviews the manufacturing process of cork stoppers, the diversity of microorganisms associated with cork, and finally the diversity and origins of the compounds associated with cork taint in wine, focusing on those currently considered as more important. Some important results recently obtained by the authors are also included. The current idea of suppressing mold growth during cork stopper manufacturing is discussed, as well as the erroneous idea of imputing, directly and exclusively, to cork the responsibility of the so-called cork taint in wine. PMID- 11052648 TI - Biodegradation of methyl tertiary-butyl ether (MTBE) and other fuel oxygenates. AB - Methyl tertiary-butyl ether (MTBE) has been added to gasoline in the U.S. for the last decade in order to meet the mandates of the 1990 Clean Air Act. This law decreed that gasoline sold in many locations must contain oxygenates to improve combustion and minimize air pollution. Unfortunately, this widespread use has led to the contamination of some soils and aquifers, and remediation is now required. Bioremediation has proven to be an environmentally responsible and cost-effective approach to remediating petroleum spills; this article reviews the potential that bioremediation may also be appropriate for remediating MTBE contamination. There is now good evidence that MTBE can be degraded by bacteria and fungi under aerobic conditions, and promising indications that the process also occurs under methanogenic and ferric iron-reducing conditions. Yet, apparently it is not a widespread phenomenon. The challenge is to find effective bioremediation strategies that maximize this biodegradation so that it can be used reliably in cleaning contaminated sites. Both simple biostimulation and more complex bioaugmentation protocols are being developed to meet this pressing need. PMID- 11052649 TI - Iraq's biological warfare agents: a comprehensive analysis. PMID- 11052650 TI - Being prepared for class: challenging taken-for-granted assumptions. PMID- 11052651 TI - Statewide nursing articulation model design: politics or academics? AB - Designing statewide nursing articulation models is more about politics than it is about academics. Resolution of the articulation question lies in collaboration, establishment of trust, dialogue, and compromise that identifies a common ground tolerable for all stakeholders in the region or state. Achieving satisfactory articulation outcomes frequently involves negotiation of conflicts, assumptions, beliefs, goals, and sometimes program survival. This article reviews factors that influence the process of articulation model design, describes steps in the process, discusses issues essential for successful implementation, and briefly describes research evaluating the effectiveness of statewide or regional articulation models. The nursing profession continues to face many workforce issues and must engage in effective strategic planning to achieve an adequate workforce. One important strategy is to build a strong, flexible educational infrastructure that includes a quality, efficient statewide articulation that enhances educational mobility for nurses with diverse backgrounds and levels of preparation. PMID- 11052652 TI - Performance expectations of the associate degree nurse graduate within the first six months. AB - The Oklahoma Associate Degree Nursing Directors Council was determined to be proactive in defining the role of the associate degree registered nurse (ADN) within the evolving health care delivery system. A task force was formed by the Council to design and implement strategies for defining the emerging roles. Strategies included surveying health care employers on the performance expectations of the ADN graduates within the first six months of employment. The employers were asked to determine the importance of various functions and abilities that the ADN graduates may or may not possess. Surveys returned were representative of a variety of rural and urban agencies throughout the state and bordering areas. Responses were grouped into 21 categories. Ranked highest was the ability to demonstrate verbal and written communication effectively with accountability to the employer and personal attributes such as open-mindedness, flexibility, and receptiveness to criticism ranked second and third. The lowest rankings received were for competency with fiscal management. The results of the survey highlight the continued need to stress strong communication skills, as well as professional accountability throughout the educational process. PMID- 11052653 TI - Health habits of nursing versus non-nursing students: a longitudinal study. AB - As our culture shifts from a sickness repair system to a health promotion and disease prevention system, nurses need to take more responsibility for practicing positive health behaviors. The problem addressed in this study was "Does exposure to nursing theory content and client interactions make any difference in the regular practice of positive health behaviors in nursing students when compared to non-nursing students?" The purpose of this study was to determine if nursing students practice healthy life styles that would help prepare them to be effective advocates for health promotion and disease prevention. The Health Habits Inventory (HHI) was used in this two-year longitudinal study to compare health habits between 71 nursing and 83 non-nursing students. There was a statistically significant difference between nursing and non-nursing students in time 1 (t = 4.91, p < .001) and time 2 (t = 3.59, p < .001) with nursing students scoring higher in health habits. Nursing students improved significantly from time 1 to time 2 (t = 2.05, p = .021) whereas nonnursing students did not improve (t = .94, p = .175). In specific behaviors, nursing students improved in eating breakfast regularly, performing monthly self breast and testicular exams, reading food labels, wearing seatbelts, and exercising at least three times a week. Implications include the importance of emphasizing self health care in nursing curricula to promote healthy life styles of nursing students who can subsequently become role models in their professional practice. PMID- 11052654 TI - Factors that predict success on the NCLEX-PN. PMID- 11052655 TI - Baccalaureate nursing students' reflections on a nontraditional mental health experience: learning outcomes. PMID- 11052656 TI - The experience of choosing nursing as a career. PMID- 11052657 TI - A pilot study exploring the perceptions of ADN graduates on their undergraduate gerontological nursing course and their nursing jobs. PMID- 11052658 TI - The effect of changes in test item sequence on student performance in multiple choice tests for baccalaureate nursing students. PMID- 11052659 TI - Using focus groups to evaluate the impact of a masters in nursing distance education program. PMID- 11052660 TI - Getting your nursing program in the "news". AB - Getting in the news usually needs to be a planned event with intentional actions and strategies. In most cases, news from the nursing program needs to be an administrative or faculty goal with a strategy for how this will occur. There must be a commitment to ensure that timely information is provided so that a news item can actually become reality. Most importantly, when newspapers and the media are provided with this information in a news release it must be written in a style that piques the interest of the editor and causes it to be selected as news (Perloff, 1997). If time is taken to interact with reporters and to understand how the media works, it can provide support to achieve the goals and outcomes of a nursing program. PMID- 11052661 TI - Heme structure of hemoglobin M Iwate [alpha 87(F8)His-->Tyr]: a UV and visible resonance Raman study. AB - Heme structures of a natural mutant hemoglobin (Hb), Hb M Iwate [alpha87(F8)His- >Tyr], and protonation of its F8-Tyr were examined with the 244-nm excited UV resonance Raman (UVRR) and the 406.7- and 441.6-nm excited visible resonance Raman (RR) spectroscopy. It was clarified from the UVRR bands at 1605 and 1166 cm(-)(1) characteristic of tyrosinate that the tyrosine (F8) of the abnormal subunit in Hb M Iwate adopts a deprotonated form. UV Raman bands of other Tyr residues indicated that the protein takes the T-quaternary structure even in the met form. Although both hemes of alpha and beta subunits in metHb A take a six coordinate (6c) high-spin structure, the 406.7-nm excited RR spectrum of metHb M Iwate indicated that the abnormal alpha subunit adopts a 5c high-spin structure. The present results and our previous observation of the nu(Fe)(-)(O(tyrosine)) Raman band [Nagai et al. (1989) Biochemistry 28, 2418-2422] have proved that F8 tyrosinate is covalently bound to Fe(III) heme in the alpha subunit of Hb M Iwate. As a result, peripheral groups of porphyrin ring, especially the vinyl and the propionate side chains, were so strongly influenced that the RR spectrum in the low-frequency region excited at 406.7 nm is distinctly changed from the normal pattern. When Hb M Iwate was fully reduced, the characteristic UVRR bands of tyrosinate disappeared and the Raman bands of tyrosine at 1620 (Y8a), 1207 (Y7a), and 1177 cm(-)(1) (Y9a) increased in intensity. Coordination of distal His(E7) to the Fe(II) heme in the reduced alpha subunit of Hb M Iwate was proved by the observation of the nu(Fe)(-)(His) RR band in the 441.6-nm excited RR spectrum at the same frequency as that of its isolated alpha chain. The effects of the distal-His coordination on the heme appeared as a distortion of the peripheral groups of heme. A possible mechanism for the formation of a Fe(III) tyrosinate bond in Hb M Iwate is discussed. PMID- 11052662 TI - Alignment of lysine-anchored membrane peptides under conditions of hydrophobic mismatch: a CD, 15N and 31P solid-state NMR spectroscopy investigation. AB - The secondary structure and alignment of hydrophobic model peptides in phosphatidylcholine membranes were investigated as a function of hydrophobic mismatch by CD and oriented proton-decoupled (15)N solid-state NMR spectroscopies. In addition, the macroscopic phase and the orientational order of the phospholipid headgroups was analyzed by proton-decoupled (31)P NMR spectroscopy. Both, variations in the composition of the polypeptide (10-30 hydrophobic residues) as well as the fatty acid acyl chain of the phospholipid (10-22 carbons) were studied. At lipid-to-peptide ratios of 50, the peptides adopt helical conformations and bilayer macroscopic phases are predominant. The peptide and lipid maintain much of their orientational order even when the peptide is calculated to be 3 A too short or 14 A too long to fit into the pure lipid bilayer. A continuous decrease in the (15)N chemical shift obtained from transmembrane peptides in oriented membranes suggests an increasing helical tilt angle when the membrane thickness is reduced. This response is, however, insufficient to account for the full hydrophobic mismatch. When the helix is much too long to span the membrane, both the lipid and the peptide order are perturbed, an indication of changes in the macroscopic properties of the membrane. In contrast, sequences that are much too short show little effect on the phospholipid headgroup order, but the peptides exhibit a wide range of orientational distributions predominantly close to parallel to the membrane surface. A thermodynamic formalism is applied to describe the two-state equilibrium between in-plane and transmembrane peptide orientations. PMID- 11052663 TI - Crystal structure of oxidized Bacillus pasteurii cytochrome c553 at 0.97-A resolution. AB - This article reports the first X-ray structure of the soluble form of a c-type cytochrome isolated from a Gram-positive bacterium. Bacillus pasteurii cytochrome c(553), characterized by a low reduction potential and by a low sequence homology with cytochromes from Gram-negative bacteria or eukaryotes, is a useful case study for understanding the structure-function relationships for this class of electron-transfer proteins. Diffraction data on a single crystal of cytochrome c(553) were obtained using synchrotron radiation at 100 K. The structure was determined at 0.97-A resolution using ab initio phasing and independently at 1.70 A in an MAD experiment. In both experiments, the structure solution exploited the presence of a single Fe atom as anomalous scatterer in the protein. For the 0.97 A data, the phasing was based on a single data set. This is the most precise structure of a heme protein to date. The crystallized cytochrome c(553) contains only 71 of the 92 residues expected from the intact protein sequence, lacking the first 21 amino acids at the N-terminus. This feature is consistent with previous evidence that this tail, responsible for anchoring the protein to the cytoplasm membrane, is easily cleaved off during the purification procedure. The heme prosthetic group in B. pasteurii cytochrome c(553) is surrounded by three alpha helices in a compact arrangement. The largely exposed c-type heme group features a His-Met axial coordination of the Fe(III) ion. The protein is characterized by a very asymmetric charge distribution, with the exposed heme edge located on a surface patch devoid of net charges. A structural search of a representative set of protein structures reveals that B. pasteurii cytochrome c(553) is most similar to Pseudomonas cytochromes c(551), followed by cytochromes c(6), Desulfovibrio cytochrome c(553), cytochromes c(552) from thermophiles, and cytochromes c from eukaryotes. Notwithstanding a low sequence homology, a structure-based alignment of these cytochromes shows conservation of three helical regions, with different additional secondary structure motifs characterizing each protein. In B. pasteurii cytochrome c(553), these motifs are represented by the shortest interhelix connecting fragments observed for this group of proteins. The possible relationships between heme solvent accessibility and the electrochemical reduction potential are discussed. PMID- 11052664 TI - Evidence that familial hypercholesterolemia mutations of the LDL receptor cause limited local misfolding in an LDL-A module pair. AB - Mutations at conserved sites within the ligand-binding LDL-A modules of the LDL receptor cause the genetic disease familial hypercholesterolemia (FH), and several of these FH mutations in modules five and six prevent the isolated single modules from folding properly to a nativelike three-dimensional structure. Because LDL-A modules occur as a series of contiguous repeats in the LDLR and related proteins, we investigated the impact of two FH mutations in LDL-A module five (D203G and D206E) and two mutations in module six (E219K and D245E) in the context of the covalently connected module five-six pair. HPLC chromatography of the products formed under conditions that efficiently refold the native module five-six pair demonstrate that, for each mutation, a folding defect persists in the module pair. NMR spectroscopy and calcium affinity measurements of the ensemble of misfolded products demonstrate that the unaltered module of each pair can fold to its native structure regardless of the range of misfolded conformations adopted by its mutated neighbor. These findings lend additional support to a model in which individual LDL-A modules of the LDL receptor act as independent structural elements. PMID- 11052665 TI - Ancient adaptation of the active site of tryptophanyl-tRNA synthetase for tryptophan binding. AB - The amino acid binding domains of the tryptophanyl (TrpRS)- and tyrosyl-tRNA synthetases (TyrRS) of Bacillus stearothermophilus are highly homologous. These similarities suggest that conserved residues in TrpRS may be responsible for both determining tryptophan recognition and discrimination against tyrosine. This was investigated by the systematic mutation of TrpRS residues based upon the identity of homologous positions in TyrRS. Of the four residues which interact directly with the aromatic side chain of tryptophan (Phe5, Met129, Asp132, and Val141) replacements of Asp132 led to significant changes in the catalytic efficiency of Trp aminoacylation (200-1250-fold reduction in k(cat)/K(M)) and substitution of Val141 by the larger Glu side chain reduced k(cat)/K(M) by 300-fold. Mutation of Pro127, which determines the position of active-site residues, did not significantly effect Trp binding. Of the mutants tested, D132N TrpRS also showed a significant reduction in discrimination against Tyr, with Tyr acting as a competitive inhibitor but not a substrate. The analogous residue in B. stearothermophilusTyrRS (Asp176) has also been implicated as a determinant of amino acid specificity in earlier studies [de Prat Gay, G., Duckworth, H. W., and Fersht, A. R. (1993) FEBS Lett. 318, 167-171]. This striking similarity in the function of a highly conserved residue found in both TrpRS and TyrRS provides mechanistic support for a common origin of the two enzymes. PMID- 11052666 TI - Dissociative mechanism of thermal denaturation of rabbit skeletal muscle glycogen phosphorylase b. AB - The thermal stability of rabbit skeletal muscle glycogen phosphorylase b was characterized using enzymological inactivation studies, differential scanning calorimetry, and analytical ultracentrifugation. The results suggest that denaturation proceeds by the dissociative mechanism, i.e., it includes the step of reversible dissociation of the active dimer into inactive monomers and the following step of irreversible denaturation of the monomer. It was shown that glucose 1-phosphate (substrate), glucose (competitive inhibitor), AMP (allosteric activator), FMN, and glucose 6-phosphate (allosteric inhibitors) had a protective effect. Calorimetric study demonstrates that the cofactor of glycogen phosphorylase-pyridoxal 5'-phosphate-stabilizes the enzyme molecule. Partial reactivation of glycogen phosphorylase b preheated at 53 degrees C occurs after cooling of the enzyme solution to 30 degrees C. The fact that the rate of reactivation decreases with dilution of the enzyme solution indicates association of inactive monomers into active dimers during renaturation. The allosteric inhibitor FMN enhances the rate of phosphorylase b reactivation. PMID- 11052667 TI - Localization of the binding site for the oligosaccharide moiety of Gb3 on verotoxin 1 using NMR residual dipolar coupling measurements. AB - By use of NMR residual dipolar coupling measurements in a dilute liquid crystalline solvent, the solution structure has been determined of the complex between the oligosaccharide moiety of globotriaosylceramide (Gb(3)-OS) and the B subunit homopentamer of verotoxin 1 (VTB). The dipolar coupling data indicate that Gb(3)-OS binds in a single binding site per monomer, which is identical to one of three sites inferred from the X-ray structure of the same complex. We find no evidence within experimental error for occupancy at either of the two additional binding sites observed per monomer in the crystal structure. PMID- 11052668 TI - Rapid phosphotransfer to CheY from a CheA protein lacking the CheY-binding domain. AB - The histidine protein kinase CheA plays a central role in the bacterial chemotaxis signal transduction pathway. Autophosphorylated CheA passes its phosphoryl group to CheY very rapidly (k(cat) approximately 750 s(-)(1)). Phospho CheY in turn influences the direction of flagellar rotation. The autophosphorylation site of CheA (His(48)) resides in its N-terminal P1 domain. The adjacent P2 domain provides a high-affinity binding site for CheY, which might facilitate the phosphotransfer reaction by tethering CheY in close proximity to the phosphodonor located in P1. To explore the contribution of P2 to the CheA --> CheY phosphotransfer reaction in the Escherichia coli chemotaxis system, we examined the transfer kinetics of a mutant CheA protein (CheADeltaP2) in which the 98 amino acid P2 domain had been replaced with an 11 amino acid linker. We used rapid-quench and stopped-flow fluorescence experiments to monitor phosphotransfer to CheY from phosphorylated wild-type CheA and from phosphorylated CheADeltaP2. The CheADeltaP2 reaction rates were significantly slower and the K(m) value was markedly higher than the corresponding values for wild-type CheA. These results indicate that binding of CheY to the P2 domain of CheA indeed contributes to the rapid kinetics of phosphotransfer. Although phosphotransfer was slower with CheADeltaP2 (k(cat)/K(m) approximately 1.5 x 10(6) M(-)(1) s(-)(1)) than with wild-type CheA (k(cat)/K(m) approximately 10(8) M(-)(1) s(-)(1)), it was still orders of magnitude faster than the kinetics of CheY phosphorylation by phosphoimidazole and other small molecule phosphodonors (k(cat)/K(m) approximately 5-50 M(-)(1) s(-)(1)). We conclude that the P1 domain of CheA also makes significant contributions to phosphotransfer rates in chemotactic signaling. PMID- 11052669 TI - Structural and functional implications of tau hyperphosphorylation: information from phosphorylation-mimicking mutated tau proteins. AB - Abnormal tau-immunoreactive filaments are a hallmark of tauopathies, including Alzheimer's disease (AD). A higher phosphorylation ("hyperphosphorylation") state of tau protein may represent a critical event. To determine the potential role of tau hyperphosphorylation in these disorders, mutated tau proteins were produced where serine/threonine residues known to be highly phosphorylated in tau filaments isolated from AD patients were substituted for glutamate to simulate a paired helical filament (PHF)-like tau hyperphosphorylation. We demonstrate that, like hyperphosphorylation, glutamate substitutions induce compact structure elements and SDS-resistant conformational domains in tau protein. Hyperphosphorylation-mimicking glutamate-mutated tau proteins display a complete functional loss in its ability to promote microtubule nucleation which can partially be overcome by addition of the osmolyte trimethylamine N-oxide (TMAO), which is similar to phosphorylated tau. In addition, glutamate-mutated tau proteins fail to interact with the dominant brain protein phosphatase 2A isoform ABalphaC, and exhibit a reduced ability to assemble into filaments. Interestingly, wild-type tau and phosphorylation-mimicking tau similarly bind to microtubules when added alone, but the mutated tau is almost completely displaced from the microtubule surface by equimolar concentrations of wild-type tau. The data indicate that glutamate-mutated tau proteins provide a useful model for analyzing the functional consequences of tau hyperphosphorylation. They suggest that several mechanisms contribute to the abnormal tau accumulation observed during tauopathies, in particular a selective displacement of hyperphosphorylated tau from microtubules, a functional loss in promoting microtubule nucleation, and a failure to interact with phosphatases. PMID- 11052670 TI - Interdependence of profilin, cation, and nucleotide binding to vertebrate non muscle actin. AB - The interaction of profilin and non-muscle beta,gamma-actin prepared from bovine spleen has been investigated under physiologic ionic conditions. Profilin binding to actin decreases the affinity of actin for MgADP and MgATP by about 65- and 13 fold, respectively. Kinetic measurements indicate that profilin binding to actin weakens the affinity of actin for nucleotides primarily due to an increased nucleotide dissociation rate constant, but the nucleotide association rate constant is also increased about 2-fold. Removal of the actin-bound nucleotide and divalent cation produces the labile intermediate species in the nucleotide exchange reaction, nucleotide free actin (NF-actin), and increases the affinity of actin for profilin about 10-fold. Profilin binds NF-actin with high affinity, K(D) = 0.013 microM, and slows the observed denaturation rate of NF-actin. Addition of ATP to NF-actin weakens the affinity for profilin and addition of Mg(2+) to ATP-actin further weakens the affinity for profilin. The high-affinity Mg(2+) of actin regulates binding of both nucleotide and profilin to actin and is important for actin interdomain coupling. The data suggest that profilin binding to actin weakens nucleotide binding to actin by disrupting Mg(2+) coordination in the actin central cleft. PMID- 11052671 TI - Evidence for a perturbation of arginine-82 in the bacteriorhodopsin photocycle from time-resolved infrared spectra. AB - Arginine-82 (R82) of bacteriorhodopsin (bR) has long been recognized as an important residue due to its absolute conservation in the archaeal rhodopsins and the effects of R82 mutations on the photocycle and proton release. However, the nature of interactions between R82 and other residues of the protein has remained difficult to decipher. Recent NMR studies showed that the two terminal nitrogens of R82 experience a highly perturbed asymmetric environment during the M state trapped at cryogenic temperatures [Petkova et al. (1999) Biochemistry 38, 1562 1572]. Although previous low-temperature FT-IR spectra of wild-type and mutant bR samples have demonstrated effects of R82 on vibrations of other amino acid side chains, no bands in these spectra were assignable to vibrations of R82 itself. We have now measured time-resolved FT-IR difference spectra of bR intermediates in the wild-type and R82A proteins, as well as in samples of the R82C mutant with and without thioethylguanidinium attached via a disulfide linkage at the unique cysteine site. Several bands in the bR --> M difference spectrum are attributable to guanidino group vibrations of R82, based on their shift upon isotope substitution of the thioethylguanidinium attached to R82C and on their disappearance in the R82A spectrum. The frequencies and intensities of these IR bands support the NMR-based conclusion that there is a significant perturbation of R82 during the bR photocycle. However, the unusually low frequencies attributable to R82 guandino group vibrations in M, approximately 1640 and approximately 1545 cm(-)(1), would require a reexamination of a previously discarded hypothesis, namely, that the perturbation of R82 involves a change in its ionization state. PMID- 11052672 TI - Reactivity of horseradish peroxidase compound II toward substrates: kinetic evidence for a two-step mechanism. AB - Transient kinetic analysis of biphasic, single turnover data for the reaction of 2,2'-azino-bis[3-ethylbenzthiazoline-6-sulfonic acid] (ABTS) with horseradish peroxidase (HRPC) compound II demonstrated preequilibrium binding of ABTS (k(+5) = 7.82 x 10(4) M(-)(1) s(-)(1)) prior to rate-limiting electron transfer (k(+6) = 42.1 s(-)(1)). These data were obtained using a stopped-flow method, which included ascorbate in the reaction medium to maintain a low steady-state concentration of ABTS (pseudo-first-order conditions) and to minimize absorbance changes in the Soret region due to the accumulation of ABTS cation radicals. A steady-state kinetic analysis of the reaction confirmed that the reduction of HRPC compound II by this substrate is rate-limiting in the complete peroxidase cycle. The reaction of HRPC with o-diphenols has been investigated using a chronometric method that also included ascorbate in the assay medium to minimize the effects of nonenzymic reactions involving phenol-derived radical products. This enabled the initial rates of o-diphenol oxidation at different hydrogen peroxide and o-diphenol concentrations to be determined from the lag period induced by the presence of ascorbate. The kinetic analysis resolved the reaction of HRPC compound II with o-diphenols into two steps, initial formation of an enzyme-substrate complex followed by electron transfer from the substrate to the heme. With o-diphenols that are rapidly oxidized, the heterolytic cleavage of the O-O bond of the heme-bound hydrogen peroxide (k(+2) = 2.17 x 10(3) s(-)(1)) is rate-limiting. The size and hydrophobicity of the o-diphenol substrates are correlated with their rate of binding to HRPC, while the electron density at the C-4 hydroxyl group predominantly influences the rate of electron transfer to the heme. PMID- 11052673 TI - Amplification of pepleomycin-mediated DNA cleavage and apoptosis by unfused aromatic cations. AB - An important approach to improve chemotherapy of members of the bleomycin (BLM) family of antibiotics is to find compounds (amplifiers) that enhance the activity of BLM-mediated DNA cleavage and apoptosis. Using a DNA-sequencing technique and pulsed field gel electrophoresis, we have investigated whether BLM-mediated cleavage of isolated and cellular DNA is amplifed by three compounds (RW-12, LS 20, 1S-5Me) which have a conformationally flexible, unfused polyaromatic system and cationic side chain in the molecules. RW-12 enhanced most effectively both pepleomycin (PEM)-induced cytotoxicity and apoptosis. The order of the maximum enhancing effect of amplifiers on PEM-mediated DNA damage is RW-12 > LS-20 > 1S 5Me. RW-12 amplified PEM-mediated DNA cleavage most effectively not only in vitro but also in cultured cells. We have reported that the order of the DNA binding constants of these compounds is RW-12 > LS-20 > 1S-5Me. In this study, we found a good correlation between PEM-mediated cleavage of isolated DNA and cellular DNA. These results suggest that BLM amplifiers bind to DNA and by doing so enhance drug-mediated DNA degradation, ultimately leading to apoptosis. The present study on amplifiers of anticancer agents shows a novel approach to the potentially effective anticancer therapy. PMID- 11052674 TI - Role of phosphatidylethanolamine in assembly and function of the factor IXa factor VIIIa complex on membrane surfaces. AB - Phospholipid membranes play a significant role during the proteolytic activation of blood coagulation proteins. This investigation identifies a role for phosphatidylethanolamine (PE) during the activation of factor X by the tenase complex, an enzymatic complex composed of the serine protease, factor IXa, a protein cofactor, factor VIIIa, a phospholipid membrane, and Ca(2+). Phospholipid vesicles composed of PE, phosphatidylserine (PS), and phosphatidylcholine support factor Xa generation. The K(m) and k(cat) for factor X activation by the tenase complex are independent of PE in the presence of 20% PS. At lower PS concentrations, the presence of 20 or 35% PE lowers the K(m) and increases the k(cat) as compared to those in vesicles without PE. The effect of PE on the k(cat) of the tenase complex is mediated through factor VIIIa. PE also enhances factor Xa generation by facilitating tenase complex formation; PE lowers the K(d(app)) of factor IXa for both phospholipid/Ca(2+) and phospholipid/Ca(2+)/factor VIIIa complexes in the presence of suboptimal PS. In addition, the K(d)s of factor IXa and factor X were lower for phospholipid vesicles containing PE. N-Methyl-PE increased the k(cat) and decreased the K(d(app)), whereas N,N-dimethyl-PE had no effect on either parameter, indicating the importance of headgroup size. Lyso-PE had no effect on kinetic parameters, indicating the sn-2 acyl chain dependence of the PE effect. Together, these results demonstrate a role for PE in the assembly and activity of the tenase complex and further extend the understanding of the importance of PE-containing membranes in hemostasis. PMID- 11052675 TI - Biosynthesis of triphosphoribosyl-dephospho-coenzyme A, the precursor of the prosthetic group of malonate decarboxylase. AB - Malonate decarboxylase from Klebsiella pneumoniae consists of four subunits MdcA, D, E, and C and catalyzes the cleavage of malonate to acetate and CO(2). The smallest subunit MdcC is an acyl carrier protein to which acetyl and malonyl thioester residues are bound via a 2'-(5' '-phosphoribosyl)-3'-dephospho-CoA prosthetic group and turn over during the catalytic mechanism. We report here on the biosynthesis of holo acyl carrier protein from the unmodified apoprotein. The prosthetic group biosynthesis starts with the MdcB-catalyzed condensation of dephospho-CoA with ATP to 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA. In this reaction, a new alpha (1' ' --> 2') glycosidic bond between the two ribosyl moieties is formed, and thereby, the adenine moiety of ATP is displaced. MdcB therefore is an ATP:dephospho-CoA 5'-triphosphoribosyl transferase. The second protein involved in holo ACP synthesis is MdcG. This enzyme forms a strong complex with the 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA prosthetic group precursor. This complex, called MdcG(i), is readily separated from free MdcG by native polyacrylamide gel electrophoresis. Upon incubation of MdcG(i) with apo acyl carrier protein, holo acyl carrier protein is synthesized by forming the phosphodiester bond between the 2'-(5' '-phosphoribosyl)-3'-dephospho-CoA prosthetic group and serine 25 of the protein. MdcG corresponds to a 2'-(5' ' triphosphoribosyl)-3'-dephospho-CoA:apo ACP 2'-(5' '-phosphoribosyl)-3'-dephospho CoA transferase. In absence of the prosthetic group precursor, MdcG catalyzes at a low rate the adenylylation of apo acyl carrier protein using ATP as substrate. The adenylyl ACP thus formed is an unphysiological side product and is not involved in the biosynthesis of holo ACP. The 2'-(5' '-triphosphoribosyl)-3' dephospho-CoA precursor of the prosthetic group has been purified and its identity confirmed by mass spectrometry and enzymatic analysis. PMID- 11052676 TI - Identification of the active site of phosphoribosyl-dephospho-coenzyme A transferase and relationship of the enzyme to an ancient class of nucleotidyltransferases. AB - Malonate decarboxylase from Klebsiella pneumoniae contains an acyl carrier protein (MdcC) to which a 2'-(5' '-phosphoribosyl)-3'-dephospho-CoA prosthetic group is attached via phosphodiester linkage to serine 25. We have shown in the preceding paper in this issue that the formation of this phosphodiester bond is catalyzed by a phosphoribosyl-dephospho-coenzyme A transferase MdcG with the substrate 2'-(5' '-triphosphoribosyl)-3'-dephospho-CoA that is synthesized from ATP and dephospho-coenzyme A by the triphosphoribosyl transferase MdcB. The reaction catalyzed by MdcG is related to nucleotidyltransfer reactions, and the enzyme indeed catalyzes unphysiological nucleotidyltransfer, e.g., adenylyltransfer from ATP to apo acyl carrier protein (ACP). These unspecific side reactions are favored at high Mg(2+) concentrations. A sequence motif including D134 and D136 of MdcG is a signature of all nucleotidyltransferases. It is known from the well-characterized mammalian DNA polymerase beta that this motif is at the active site of the enzyme. Site-directed mutagenesis of D134 and/or D136 of MdcG to alanine abolished the transfer of the prosthetic group to apo ACP, but the binding of triphosphoribosyl-dephospho-CoA to MdcG was not affected. Evidence is presented that similar to MdcG, MadK encoded by the malonate decarboxylase operon of Malonomonas rubra and CitX from the operon encoding citrate lyase in Escherichia coli are phosphoribosyl-dephospho-CoA transferases catalyzing the attachment of the phosphoribosyl-dephospho-CoA prosthetic group to their specific apo ACPs. PMID- 11052677 TI - Raman spectroscopy of uracil DNA glycosylase-DNA complexes: insights into DNA damage recognition and catalysis. AB - Using off-resonance Raman spectroscopy, we have examined each complex along the catalytic pathway of the DNA repair enzyme uracil DNA glycosylase (UDG). The binding of undamaged DNA to UDG results in decreased intensity of the DNA Raman bands, which can be attributed to an increased level of base stacking, with little perturbation in the vibrational modes of the DNA backbone. A specific complex between UDG and duplex DNA containing 2'-beta-fluorodeoxyuridine shows similar increases in the level of DNA base stacking, but also a substrate directed conformational change in UDG that is not observed with undamaged DNA, consistent with an induced-fit mechanism for damage site recognition. The similar increases in the level of DNA base stacking for the nonspecific and specific complexes suggest a common enzyme-induced distortion in the DNA, potentially DNA bending. The difference spectrum of the extrahelical uracil base in the substrate analogue complexes reveals only a small electron density reorganization in the uracil ring for the ground state complex, but large 34 cm(-)(1) downshifts in the carbonyl normal modes. Thus, UDG activates the uracil ring in the ground state mainly through H bonds to its C=O groups, without destroying its quasi aromaticity. This result is at variance with the conclusion from a recent crystal structure, in which the UDG active site significantly distorts the flipped-out pseudouridine analogue such that a change in hybridization at C1 occurs [Parikh, S. S., et al. (2000) Proc. Natl. Acad. Sci. USA 97, 5083]. The Raman vibrational signature of the bound uracil product differs significantly from that of free uracil at neutral pH, and indicates that the uracil is anionic. This is consistent with recent NMR results, which established that the enzyme stabilizes the uracil anion leaving group by 3.4 pK(a) units compared to aqueous solution, contributing significantly to catalysis. These observations are generally not apparent from the high-resolution crystal structures of UDG and its complexes with DNA; thus, Raman spectroscopy can provide unique and valuable insights into the nature of enzyme-DNA interactions. PMID- 11052678 TI - Determination of the binding specificity of the SH2 domains of protein tyrosine phosphatase SHP-1 through the screening of a combinatorial phosphotyrosyl peptide library. AB - A method for the rapid identification of high-affinity ligands to Src homology-2 (SH2) domains is reported. A phosphotyrosyl (pY) peptide library containing completely randomized residues at positions -2 to +3 relative to the pY was synthesized on TentaGel resin, with a unique peptide sequence on each resin bead (total 2.5 x 10(6) different sequences). The library was screened against the biotinylated N- and C-terminal SH2 domains of protein tyrosine phosphatase SHP-1, and the beads that carry high-affinity ligands of the SH2 domains were identified using an enzyme-linked assay involving a streptavidin-alkaline phosphatase conjugate. Peptide ladder sequencing of the selected beads using matrix-assisted laser desorption ionization mass spectrometry revealed consensus sequences for both SH2 domains. The N-terminal SH2 domain strongly selects for peptides with a leucine at the -2 position; at the C-terminal side of the pY residue, it can recognize two distinct classes of peptides with consensus sequences of LXpY(M/F)X(F/M) and LXpYAXL (X = any amino acid), respectively. The C-terminal SH2 domain exhibits almost exclusive selectivity for peptides of the consensus sequence, (V/I/L)XpYAX(L/V). Several representative sequences selected from the library were individually synthesized and tested for binding to the SH2 domains by surface plasmon resonance and for their ability to stimulate the catalytic activity of SHP-1. Both experiments have demonstrated that the selected peptides are capable of binding to the SH2 domains with dissociation constants (K(D)) in the low micromolar range. PMID- 11052679 TI - Enzyme inhibition assays using fluorescence correlation spectroscopy: a new algorithm for the derivation of kcat/KM and Ki values at substrate concentrations much lower than the Michaelis constant. AB - A new mathematical formalism is deduced which allows for the calculation of the k(cat) over K(M) ratio based on measurements of the enzyme kinetics with substrate concentrations much lower than K(M). The equations are also applied on the action of an inhibitor on enzyme activity yielding the binding constant, K(i), of an inhibitor molecule. For practical evaluation of the new theoretical approach, the enzymatic reaction of CD45 phosphatase was used as a well characterized model system with known inhibitors for testing the K(i) value determination scheme. The k(cat)/K(M) ratio was calulated to be 4.7 x 10(5) M( )(1) s(-)(1), the K(i) of the inhibitor molecule PKF52-524 was estimated to be (1 2) x 10(-)(7) M and the association rate of the inhibitor PKF52-524 to CD45 phosphatase was estimated to be 59 M(-)(1) s(-)(1). PMID- 11052680 TI - Molecular structure of a fibrillar Alzheimer's A beta fragment. AB - Amyloid-beta (Abeta) peptide deposition as fibrillar senile plaques is a key element in the pathology of Alzheimer's disease. Here we present a high resolution structure of an Abeta amyloid fibril using magnetically aligned preparations of a central Abeta domain which forms representative amyloid fibrils. Diffraction analysis of these samples revealed Bragg reflections on layer lines consistent with a preferred orientation, as opposed to the typical symmetry associated with fibers. These crystalline properties permitted a molecular replacement approach based upon a beta-hairpin motif resulting in a structure of the fibrillar Abeta peptide. This detailed molecular structure of Abeta in its fibrous state provides clues as to the mechanism of amyloid assembly and identifies potential targets for controlling the aggregation process. PMID- 11052681 TI - Catalytic assessment of the glycine-rich loop of the v-Fps oncoprotein using site directed mutagenesis. AB - The three glycine residues in the glycine-rich loop of the oncoprotein, v-Fps, were mutated to determine the function of these highly conserved residues in catalysis. The kinase domains of six mutants (G928A,S, G930A,S, and G933A,S) and the wild-type enzyme were expressed and purified as fusion proteins of glutathione-S-transferase in Escherichia coli, and their catalytic properties were assessed using steady-state kinetic, inhibition, viscosity and autophosphorylation studies. Although both G928A and G930A had no detectable activity toward the substrate peptide (EAEIYEAIE), the other mutants had apparent, but varying activities. G930S lowered the rate of phosphoryl transfer by 130-fold while G928S and G933S had smaller (6-9-fold) reductions in this step. These effects on catalytic function parallel the reductions in turnover and autophosphorylation but, for G933S and G933A, net product release is still rate limiting at saturating substrate and ATP concentrations. On the basis of K(I) measurements, the effects on turnover for these mutants may be due to improved ADP affinity. While ADP affinity is reduced 2- and 3-fold for G928S and G930S, the affinity of this product is increased by 22- and 7-fold for G933S and G933A. In contrast, ATP affinity is enhanced by 5-fold for G928S and G933S and is reduced by less than 2-fold for G930S. These complex, differential effects on nucleotide binding indicate that the glycines influence the relative affinities of ADP and ATP. On the basis of the results of serine replacements, Gly-928 and Gly-930 enhance ADP affinity by 9- and 2-fold compared to ATP affinity whereas Gly-933 diminishes ADP affinity by approximately 4-fold compared to ATP affinity. These findings demonstrate that the functions of the loop lie not only in modulating the rate of the phosphoryl transfer step but also in balancing the relative affinities of ATP and ADP. These effects on nucleotide specificity may be a contributing element for the stabilization of the phosphoryl transition state and may also facilitate quick release of bound products. PMID- 11052682 TI - Enhancement of the enzymatic activity of ribonuclease HI from Thermus thermophilus HB8 with a suppressor mutation method. AB - A genetic method for isolating a mutant enzyme of ribonuclease HI (RNase HI) from Thermus thermophilus HB8 with enhanced activity at moderate temperatures was developed. T. thermophilus RNase HI has an ability to complement the RNase H dependent temperature-sensitive (ts) growth phenotype of Escherichia coli MIC3001. However, this complementation ability was greatly reduced by replacing Asp(134), which is one of the active site residues, with His, probably due to a reduction in the catalytic activity. Random mutagenesis of the gene encoding the resultant D134H enzyme, followed by screening for second-site revertants, allowed us to isolate three single mutations (Ala(12) --> Ser, Lys(75) --> Met, and Ala(77) --> Pro) that restore the normal complementation ability to the D134H enzyme. These mutations were individually or simultaneously introduced into the wild-type enzyme, and the kinetic parameters of the resultant mutant enzymes for the hydrolysis of a DNA-RNA-DNA/DNA substrate were determined at 30 degrees C. Each mutation increased the k(cat)/K(m) value of the wild-type enzyme by 2.1-4.8 fold. The effects of the mutations on the enzymatic activity were roughly cumulative, and the combination of these three mutations increased the k(cat)/K(m) value of the wild-type enzyme by 40-fold (5.5-fold in k(cat)). Measurement of thermal stability of the mutant enzymes with circular dichroism spectroscopy in the presence of 1 M guanidine hydrochloride and 1 mM dithiothreitol showed that the T(m) value of the triple mutant enzyme, in which all three mutations were combined, was comparable to that of the wild-type enzyme (75.0 vs 77.4 degrees C). These results demonstrate that the activity of a thermophilic enzyme can be improved without a cost of protein stability. PMID- 11052683 TI - HAI-RTP determination of carbaryl pesticide in different irrigation water samples of south Spain. AB - In this work, a widely used pesticide named carbaryl, present in numerous water supplies, has been determined by the so-called heavy atom induced room temperature phosphorescence (HAI-RTP) methodology. A detailed study of numerous instrumental variables such as sensitivity, slits, decay time, and gate time, and those of experimental type such as heavy atoms, oxygen scavenger, temperature, buffer solutions, and organic solvents, have been carried out. The detection limit was 2.8 ng mL(-)(1) with a relative standard deviation of 2.12% at the 150 ng mL(-)(1) level. Spiked irrigation water samples taken from different places near agricultural fields gave mean percentage recoveries of 95.7%. The results obtained in this study indicate that the proposed method is suitable for the determination of residues of carbaryl pesticide in water samples with good reproducibility and sensitivity for the analysis of this compound being rapid and very simple for routine analysis. PMID- 11052684 TI - Contamination of Italian citrus essential oils: presence of chloroparaffin. AB - In this study, the contamination by chloroparaffin of Sicilian and Calabrian citrus essential oils, produced in the crop years 1994-1996, was investigated. The analyses were carried out on 102 lemon oils, 98 orange oils, and 96 mandarin oils, using a dual-channel GC-ECD. It was found that 53% of lemon oil, 33% of orange oil, and 38% of mandarin oil samples were contaminated. The mean contamination levels were 7.1 ppm (lemon), 2.5 ppm (orange), and 5.3 ppm (mandarin). The highest concentration of chloroparaffin found was 60 ppm in a lemon oil sample. PMID- 11052685 TI - Novel method for the discrimination of tuna (Thunnus thynnus) and bonito (Sarda sarda) DNA. AB - A novel method for the discrimination of bluefin tuna (Thunnus thynnus) from Atlantic bonito (Sarda sarda) was developed, based on species-specific amplification of a region of the mitochondrial cytochrome b gene by Polymerase Chain Reaction (PCR). The method, which uses a one-step amplification reaction, is more rapid to perform than any of the currently described techniques for species determination in fish. The species of origin of the DNA is indicated by the distinctive size of the PCR product on electrophoresis, but the test could readily be adapted to other forms of electrophoresis or fluorescence-based systems for quantification. Given the possibility of intraspecific variability in mitochondrial DNA and the consequent desirability of performing two independent tests, the new method constitutes a valuable addition to the range of tuna speciation methodologies currently available. PMID- 11052686 TI - Factors affecting the collection and fitting of nuclear magnetic cross-relaxation spectroscopy data with application to waxy corn starch. AB - An examination of the methods for nuclear magnetic cross-relaxation spectroscopy (CRS) data collection and analysis was conducted using water and an aqueous waxy corn starch suspension to better perform and interpret the results obtained using CRS. The CRS data collection properties evaluated were the time to achieve steady state saturation, the direct saturation of liquid protons, generation of transverse magnetization, and dependence of the offset frequency and radio frequency (RF) field strength of longitudinal relaxation in the presence of RF saturation. Effects were evaluated for variations of input values of RF saturation field strength, apparent cross-relaxation rate, and solid longitudinal relaxation rate on the results for solid content and solid internal mobility from fitting NMR data to modified theoretical expressions. Discrepancies between fitted and stoichiometric values for the solid to liquid proton ratio were investigated. The fitting procedure used a Gaussian line shape for RF saturation of the solid-like spin system and a Lorentzian line shape for RF saturation of the liquid-spin system. Conditions under which acceptable results can be obtained with limited data sets are discussed. PMID- 11052687 TI - Determination of total dietary fiber of intact cereal food products by near infrared reflectance. AB - Near-infrared reflectance spectra of cereal food products were acquired with a commercial dual-diode-array (Si, InGaAs) spectrometer customized to allow rapid acquisition of scans of intact breakfast cereals, snack foods, whole grains, and milled products. Substantial gains in the performance of multivariate calibration models generated from these data were obtained by a computational strategy that systematically analyzed the performance of various spectral windows. The calibration model based on 137 cereal food products determined the total dietary fiber (TDF) content of a test set of 45 intact diverse cereal food products with root-mean-squared error of cross-validation of between 1.8 and 2.0% TDF, relative to the laborious enzymatic-gravimetric reference method. The calibration performance is adequate to estimate TDF over the range of values found in diverse types of cereal food products (0.7-50.1%). The method requires no sample preparation and is relatively unaffected by specimen moisture content. PMID- 11052688 TI - Determination of glucosinolates in canola seeds using anion exchange membrane extraction combined with the high-pressure liquid chromatography detection. AB - A rapid, simple, and reliable method for the determination of individual glucosinolates in canola seeds was developed using a semiquantitative extraction of glucosinolates with anion exchange membranes and HPLC detection. In this one step extraction procedure, a membrane (7 cm(2)) is placed in the seed suspension prepared by grinding and boiling 0.8 g of seeds in 20 mL of water. After 10 min of shaking on the mechanical shaker, the membrane is removed from the suspension, washed, and transferred to a vial containing 5 mL of 1 N tetramethylammonium chloride. The glucosinolates are eluted from the membrane by shaking the membrane for 10 min with the eluting solvent. The glucosinolate content in membrane eluates is determined by HPLC using sinigrin standards. A coefficient of variation ranging from 1.9 to 7.6% for aliphatic glucosinolates indicated very good reproducibility of the method. Because of the instability of 4 hydroxyglucobrassicin, the coefficient of variation for the determination of this indolyl glucosinolate was 13.9%. To verify the results of the membrane extraction/HPLC detection, this new method was compared with the existing colorimetric and GC procedures. Very good correlation (R(2) = 0.98) was obtained between the total glucosinolates determined by the membrane extraction/HPLC method and the palladate colorimetric procedure for 17 canola varieties. Concentrations of individual glucosinolates in five canola varieties were compared with the GC data. Very good agreement between these two methods was obtained for aliphatic glucosinolates. However, the membrane extraction/HPLC method yielded slightly higher values for 4-hydroxyglucobrassicin than the GC method, possibly indicating that the decomposition of this glucosinolate was reduced during the sample extraction with the membranes. The simplicity and low cost of the membrane extraction/HPLC method make it an attractive alternative to the existing procedures for glucosinolate analysis in canola seeds. PMID- 11052689 TI - Hapten synthesis for the development of a competitive inhibition enzyme immunoassay for thiram. AB - An enzyme-linked immunosorbent assay (ELISA) was developed for the fungicide thiram. Two types of haptens were synthesized. The first type exhibits the two symmetrical N-alkyl dithiocarbamate patterns of thiram with a spacer arm linked to one of the N-methyl terminal group. The second type exhibits one of the two symmetrical N-alkyl dithiocarbamate patterns of thiram with a variable-length spacer arm linked to one sulfur atom. Polyclonal antibodies suitable for thiram detection were obtained from immunization with an hapten of the first type, while haptens of the second type were used as coating antigens to develop a competitive ELISA against thiram. The IC(50) value for thiram was estimated to be 0.24 microg/mL, with a detection limit of 0.03 microg/mL. The assay seems to be thiram specific since no or little cross-reaction with other dithiocarbamates were observed. PMID- 11052690 TI - Analytical method for the determination of atrazine and its dealkylated chlorotriazine metabolites in water using gas chromatography/mass selective detection. AB - A multiresidue method is reported for the determination of atrazine and its dealkylated chlorotriazine metabolites in water. Water samples are buffered to pH 10 and partitioned in ethyl acetate. Final analysis is accomplished using gas chromatography/mass selective detection (GC/MSD) in the selected ion monitoring (SIM) mode. The limit of detection (LOD) is 0.050 ng and the limit of quantification (LOQ) is 0.10 ppb for 2-chloro-4-(ethylamino)-6-(isopropylamino)-s triazine (atrazine), 2-amino-4-chloro-6-(isopropylamino)-s-triazine (G-30033), 2 amino-4-chloro-6-(ethylamino)-s-triazine (G-28279), and 2, 4-diamino-6-chloro-s triazine (G-28273). The mean procedural recoveries were 90, 92, 98, and 85% and the standard deviations were 12, 13, 16, and 20% for atrazine, G-30033, G-28279, and G-28273, respectively (n = 30). The study was conducted under U.S. EPA FIFRA Good Laboratory Practice Guidelines 40 CFR 160 for method validation. The reported procedure accounts for residues of G-28273 in water that are not included in EPA Method 507. PMID- 11052691 TI - Rapid and sensitive determination of 4-nitrophenol, 3-methyl-4-nitrophenol, 4,6 dinitro-o-cresol, parathion-methyl, fenitrothion, and parathion-ethyl by liquid chromatography with electrochemical detection. AB - Liquid chromatography with electrochemical detection has been used to determine various nitropesticides, DNOC, fenitrothion, and parathion (methyl and ethyl), and some of their main metabolites, 4-nitrophenol for parathion (methyl and ethyl) and 3-methyl-4-nitrophenol for fenitrothion, by using indirect detection. Analysis of them in river water samples has been performed without a preconcentration step. The recovery efficiencies of the tested compounds yielded values between 96 and 112% at the fortification level of 0.5 ppb in a river water sample, and their relative standard deviations were between 1 and 15%. The detection limits of these compounds ranged between 0.05 and 0.14 ppb. PMID- 11052692 TI - Continuous sorbent preconcentration for the electrothermal atomic absorption spectrometric determination of ultratrace amounts of cobalt in milled wheat fractions. AB - A method for the rapid determination of cobalt at ultratrace levels was applied in flour and flour byproducts (shorts and bran) obtained from various types of wheat that includes on-line preconcentration and electrothermal atomic absorption spectrometry is proposed. Solutions in 0.1 mol/L HNO(3) of milled wheat fractions subjected to wet ashing are preconcentrated in a straightforward flow-injection system by sorption on a RP-C(18) column following chelation. The eluent, ethanol, is carried by an air stream, and the chelate is eluted and collected in a 500 microL PTFE autosampler cup. The determination of cobalt features a precision (RSD) of 6% for a concentration of 0.2 ng/mL and a sensitivity (slope of the calibration graph) of 0.38 +/- 0.03 A s x ng/mL. The cobalt content in each type of wheat was found to be influenced by its geographical origin and texture. Also, it was found to depend on the amount of bran present in the milled wheat fraction. On the other hand, it is not significantly affected by the technological processes involved in wheat milling at a flour-producing factory. PMID- 11052693 TI - Development of enzyme-linked immunosorbent assays for toxic larkspur (Delphinium spp.) alkaloids. AB - Larkspur (Delphinium spp.) poisons thousands of cattle on western rangelands each year. Because poisoning does not cause specific lesions, and poisoned animals are rarely found before they die, definitively identifying poisoned animals is difficult. Additionally, toxin concentrations in larkspur plants vary with environment, plant, and location. Rapid, sensitive, and specific diagnostic techniques are needed to identify poisoned animals and to determine when and what plants are likely to poison livestock. In this study, three competitive inhibition enzyme-linked immunosorbent assays (CI-ELISA) for toxic larkspur alkaloids were developed. One assay is class-specific toward the N (methylsuccinimido)anthranoyllycoctonine (MSAL) alkaloids, and two assays are specific for individual alkaloids. The assay with the lowest limit of detection had an I(50) of 191 pg with a limit of detection of 30.5 pg for methyllycaconitine. Spike and recovery studies using bovine blood and brain tissue ranged from 52 to 89%. These findings suggest that with additional development these techniques are likely to be excellent tools for diagnosing poisoned animals and identifying highly toxic plants. PMID- 11052694 TI - Cleavage of the diketonitrile derivative of the herbicide isoxaflutole by extracellular fungal oxidases. AB - Isoxaflutole is a herbicide activated in soils and plants to its diketonitrile derivative, the active herbicide principle. The diketonitrile derivative undergoes cleavage to the inactive benzoic acid analogue. In this paper, it is established that an oxidative mechanism implicating two successive reactions in the presence of dimethyldioxirane can chemically initiate the cleavage of the diketonitrile. It is also shown that two white rot strains, Phanerochaete chrysosporium and Trametes versicolor, are able to convert the diketonitrile to the acid when cultured in liquid media. This main metabolite amounts to 24.6 and 15.1% of initial herbicide content after 12-15 days of culture. Another polar metabolite represents <3.7% of the parent compound amount during the same period. Oxidative enzymes produced by the fungi show a time course similar to that of diketonitrile degradation. Purified laccase (EC 1. 10.3.2), in the presence of 2 mM 2, 2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid) acting as a redox mediator at pH 3 supports the reaction with rates of 0.3-0.4 nmol h(-)(1) unit( )(1). PMID- 11052695 TI - Irreversible unfolding of myoglobin in an aqueous solution by supercritical carbon dioxide. AB - The conformational changes in myoglobin, treated by microbubbling of supercritical carbon dioxide (SC-CO(2)), were investigated by measuring the circular dichroism spectra in the ultraviolet range and compared with those in other proteins (ovoalbumin, bovine serum albumin, and beta-lactoglobulin). Irreversible unfoldings were observed after the microbubbling of SC-CO(2) at 35 degrees C and 30 MPa for 30 min. The degree of unfolding depended on the number of intramolecular S-S bonds. alpha-Helix contents of myoglobin decreased with increasing density of SC-CO(2). Unfoldings of myoglobin induced by heating, pH lowering, and the addition of a denaturant were reversible. The irreversible unfolding of myoglobin was also observed by the bubbling of gaseous CO(2) under atmospheric pressure, but heating was required. PMID- 11052696 TI - Microassay for rapid screening of alpha-amylase activity. AB - A microassay was developed for measuring the activity of alpha-amylases in the nanogram enzyme concentration range, based on the use of dye-labeled cross-linked starch as the substrate, and the release of soluble colored fragments formed in enzyme hydrolysis. Reaction conditions were optimized to generate a linear correlation between the increase in absorbance and a reaction time of 0-10 min, as well as enzyme concentrations in the range of 0-50 ng. A standard curve for the conversion of absorbance to enzyme activity units was constructed. The protocol developed was applied to monitoring the production of ultralow concentrations of recombinant barley alpha-amylase in yeast cells. PMID- 11052697 TI - Thermal dependence of malate synthase activity and its relationship to the thermal dependence of seedling emergence. AB - Cotton yields are often reduced by low temperatures at early planting dates. Improved seedling metabolism at low temperatures may enhance seedling performance. The glyoxylate cycle plays a role in the metabolism of stored lipids, and thus thermal limitations on the function of malate synthase (EC 4.1.3.2) may be involved in low-temperature limitations on seedlings. The thermal dependencies of the apparent K(M) and maximal velocity for the malate synthases from cotton (Gossypium hirsutum L.) and sunflower (Helianthus annuus L.) were determined across a 15-45 degrees C thermal range and used to estimate the thermal dependence of reaction velocity. The thermal dependence of seedling emergence was monitored for both species. The thermal dependencies of predicted reaction velocity and the measured rates of seedling emergence are correlated (cotton r(2) = 0.9, sunflower r(2) = 0.76) and suggest that the thermal dependencies of enzymes predicted from basic kinetic parameters may be useful indicators of the thermal dependence of more complex whole-plant processes. PMID- 11052698 TI - Conversion to docosahexaenoic acid-containing phosphatidylserine from squid skin lecithin by phospholipase D-mediated transphosphatidylation. AB - Phospholipase D (PLD)-mediated transphosphatidylation of squid skin lecithin with L-serine was examined to prepare docosahexaenoic acid-containing phosphatidylserine (DHA-PS). When a biphasic system with organic solvent and 0.2 M acetate buffer (pH 5.5) was used, PS synthesis was significantly affected by the amount of 3.4 M L-serine-containing acetate buffer. L-Serine concentration in the acetate buffer and choice of organic solvent were also crucial. In a typical reaction with 0.8 unit of PLD (Streptomyces sp.), 2.5 mL of ethyl acetate substrate solution containing 30 mg of squid skin lecithin in combination with 3 mL of 3.4 M L-serine-containing 0.2 M acetate buffer (pH 5.5), PS content in the recovered phospholipid fraction increased to 43.1% after 24 h. DHA composed 37.6% of fatty acids in the converted PS. This was the same DHA level as in the substrate. Phosphatidylcholine (squid skin PC, DHA 44.2%) in the squid skin lecithin was more effectively converted to PS than phosphatidylethanolamine. PMID- 11052699 TI - Immunochemical evaluation of bovine beta-casein and its 1-28 phosphopeptide in cheese during ripening. AB - Polyclonal antibodies raised against the plasmin-released 1-28 phosphopeptide from bovine beta-casein [i.e., beta-CN(f1-28)4P] specifically recognized the tryptic beta-casein 1-25 and 2-25 peptides, whatever the degree of phosphorylation, but were unresponsive to the shortened beta-casein 16-22 phosphopeptide. These antibodies were able to recognize the parent bovine beta casein as well as the homologous water buffalo protein, but they could not detect the homologous counterparts from ovine and caprine milks. Such antibodies were used in competitive enzyme-linked immunosorbent assays to monitor the plasmin mediated release of the 1-28 phosphopeptide from beta-casein and to evaluate the residual native beta-casein in bovine cheese sampled during ripening. Applications of these polyclonal antibodies are suggested mainly for estimating the age of hard cheeses and, possibly, for tracing the presence of bovine casein in fresh ovine and caprine cheeses. PMID- 11052700 TI - Antifungal and peroxidative activities of nonheme chloroperoxidase in relation to transgenic plant protection. AB - Nonheme chloroperoxidase (CPO-P) of Pseudomonas pyrrocinia catalyzes the oxidation of alkyl acids to peracids by hydrogen peroxide. Alkyl peracids possess potent antifungal activity as found with peracetate: 50% killing (LD(50)) of Aspergillus flavus occurred at 25 microM compared to 3.0 mM for the hydrogen peroxide substrate. To evaluate whether CPO-P could protect plants from fungal infection, tobacco was transformed with a gene for CPO-P from P. pyrrocinia and assayed for antifungal activity. Leaf extracts from transformed plants inhibited growth of A. flavus by up to 100%, and levels of inhibition were quantitatively correlated to the amounts of CPO-P activity expressed in leaves. To clarify if the peroxidative activity of CPO-P could be the basis for the increased resistance, the antifungal activity of the purified enzyme was investigated. The LD(50) of hydrogen peroxide combined with CPO-P occurred at 2.0 mM against A. flavus. Because this value was too small to account for the enhanced antifungal activity of transgenic plants, the kinetics of the enzyme reaction was examined and it was found that the concentration of hydrogen peroxide needed for enzyme saturation (K(m) = 5.9 mM) was already lethal. Thus, the peroxidative activity of CPO-P is not the basis for antifungal activity or enhanced resistance in transgenic plants expressing the gene. PMID- 11052701 TI - Ellagic acid, vitamin C, and total phenolic contents and radical scavenging capacity affected by freezing and frozen storage in raspberry fruit. AB - The ellagic acid, total phenolic, and vitamin C contents in four raspberry cultivars (Heritage, Autumn Bliss, Rubi, and Zeva) grown in Spain were detected and quantified by HPLC in fresh, just frozen, and stored fruits at -20 degrees C for a one year period. Ellagic acid [207-244 mg kg(-)(1) of fresh weight (fw)], total phenolic (137-1776 mg kg(-)(1) of fw), and vitamin C (221-312 mg kg(-)(1) of fw) contents in raw material were higher in the late cultivars Zeva and Rubi than in the early cultivars Autumn Bliss and Heritage. The freezing process slightly affected the values of extracted ellagic acid, total phenolic, and vitamin C content. At the end of long-term frozen storage (12 months), no significant change of total phenolic content extracted was observed, but significant decreases of 14-21% in ellagic acid and of 33-55% in vitamin C were quantified. Free radical scavenging capacity measured as antiradical efficiency (AE) depends on the seasonal period of harvest. Late cultivars, Rubi (6.1 x 10( )(4)) and Zeva (10.17 x 10(-)(4)), showed higher AE than early cultivars, Heritage (4.02 x 10(-)(4)) and Autumn Bliss (4.36 x 10(-)(4)). The freezing process produced a decrease of AE values in the four cultivars ranging between 4 and 26%. During the frozen storage, the AE values reached after the freezing process remained unchanged. PMID- 11052702 TI - Ultraviolet C irradiation at 0.5 kJ.m(-)(2) reduces decay without causing damage or affecting postharvest quality of star ruby grapefruit (C. paradisi Macf.). AB - Star Ruby grapefruit [Citrus paradisi (Macf.)] were harvested in November, February, and May, treated with ultraviolet C (UV-C) light at 0.5, 1.5, or 3.0 kJ.m(-)(2), and then stored at 7 degrees C and 90-95% relative humidity (RH) for 4 weeks with 1 additional week at 20 degrees C and approximately 80% RH. Untreated fruits were used as control. UV-C irradiation at 0.5 kJ.m(-)(2) effectively reduced decay development as compared to nontreated fruit without causing damage. Irradiation at dosages >0.5 kJ.m(-)(2) did not further improve decay control and caused rind browning and necrotic peel, the extent of damage depending on treatment dosage and harvest date. The percentage of damaged fruit after irradiation at the higher UV-C dosages was significantly higher in fruit harvested in November; differences between fruits harvested in February and May were negligible. After UV-C irradiation, the phytoalexins scoparone and scopoletin accumulated in flavedo tissue, their amounts depending on harvest date and UV-C dosage. Both phytoalexins showed similar accumulation patterns, although the concentrations of scoparone were much lower than those of scopoletin. Phytoalexin levels increased in most samples as the treatment dosage increased. No detectable levels of scoparone and scopoletin could be found in nonirradiated fruit. The influence of UV-C treatments on soluble solids concentration and titratable acidity of juice was negligible. PMID- 11052703 TI - Effect of enzymatic and chemical oxidation on the antioxidant capacity of catechin model systems and apple derivatives. AB - Changes in the chain-breaking activity of catechin model systems and apple derivatives were studied as a consequence of enzymatic and chemical oxidation. Although in different time scales, both enzymatic and chemical oxidations of catechin promoted an initial increase and a following decrease in the chain breaking activity. The latter was associated with the formation of brown pigments. Similar changes were detected during storage at -18 degrees C of nonoxidized and enzymatically oxidized apple purees. The initial increase in antioxidant activity upon oxidation was attributed to the formation of procyanidins with larger aromatic structure and to synergy effects among them. The formation of tannins, the reactivity of which is sterically hindered, was proposed as a possible explanation for the decrease in antioxidant capacity in the advanced phases of oxidation. PMID- 11052704 TI - Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing. AB - The antioxidant activity of pomegranate juices was evaluated by four different methods (ABTS, DPPH, DMPD, and FRAP) and compared to those of red wine and a green tea infusion. Commercial pomegranate juices showed an antioxidant activity (18-20 TEAC) three times higher than those of red wine and green tea (6-8 TEAC). The activity was higher in commercial juices extracted from whole pomegranates than in experimental juices obtained from the arils only (12-14 TEAC). HPLC-DAD and HPLC-MS analyses of the juices revealed that commercial juices contained the pomegranate tannin punicalagin (1500-1900 mg/L) while only traces of this compound were detected in the experimental juice obtained from arils in the laboratory. This shows that pomegranate industrial processing extracts some of the hydrolyzable tannins present in the fruit rind. This could account for the higher antioxidant activity of commercial juices compared to the experimental ones. In addition, anthocyanins, ellagic acid derivatives, and hydrolyzable tannins were detected and quantified in the pomegranate juices. PMID- 11052705 TI - Contribution of periderm material and blanching time to the quality of pasteurized peach puree. AB - Fresh peaches were blanched for either a long (20 min) or short (2 min) time, with and without periderm material. Samples were then macerated into purees and pasteurized in boiling water for 30 min. Samples were subsequently stored at 40 degrees C for 4 weeks to determine physicochemical and sensory changes affecting overall quality and nutritional content. Purees containing periderm had higher antioxidant activity (AOX) and individual phenolic acid content after processing and storage, with good correlation to AOX observed with chlorogenic acid (r = 0.82). Long blanch times resulted in increased levels of extraction and retention of both total soluble phenolics and ascorbic acid. Quantitative descriptive analysis demonstrated increased aromatic perception of skin in samples containing periderm material, but the remaining sensory attributes were indistinguishable between treatments. Macerating peaches without periderm removal was demonstrated to increase levels of bioactive phytochemicals and increase the processing yield by 7. 6%, without significantly impacting product quality, color, or sensory attributes. PMID- 11052706 TI - Effects of pulsed electric fields on the quality of orange juice and comparison with heat pasteurization. AB - Effects of pulsed electric fields (PEF) at 35 kV/cm for 59 micros on the quality of orange juice were investigated and compared with those of heat pasteurization at 94.6 degrees C for 30 s. The PEF treatment prevented the growth of microorganisms at 4, 22, and 37 degrees C for 112 days and inactivated 88% of pectin methyl esterase (PME) activity. The PEF-treated orange juice retained greater amounts of vitamin C and the five representative flavor compounds than the heat-pasteurized orange juice during storage at 4 degrees C (p < 0.05). The PEF-treated orange juice had lower browning index, higher whiteness (L), and higher hue angle (theta) values than the heat-pasteurized orange juice during storage at 4 degrees C (p < 0. 05). The PEF-treated orange juice had a smaller particle size than the heat-pasteurized orange juice (p < 0.05). degrees Brix and pH values were not significantly affected by processing methods (p > 0. 05). PMID- 11052707 TI - Effect of postharvest ultraviolet irradiation on resveratrol and other phenolics of cv. Napoleon table grapes. AB - In the skin of cv. Napoleon table grapes, the anthocyanins malvidin 3-glucoside (and its acetyl and p-coumaroyl derivatives), cyanidin 3-glucoside, peonidin 3 glucoside, cyanidin 3-glucoside, petunidin 3-glucoside, and delphinidin 3 glucoside were identified by HPLC-DAD-MS. In addition, quercetin 3-glucoside and 3-glucuronide, caffeoyltartaric, piceid, and resveratrol were also detected. The content of most phenolics remained quite constant during postharvest refrigerated storage (10 days at 0 degrees C) while the resveratrol derivatives increased 2 fold. Postharvest treatments of grapes with UVC and UVB light induced a large increase in resveratrol derivatives (3- and 2-fold, respectively). This means that a serving of mature Napoleon grapes (200 g) provides approximately 1 mg of resveratrol, which is in the range of the amount supplied by a glass of red wine. This can be increased to 2 or 3 mg of resveratrol per serving in grapes that have been irradiated with UVB or UVC, respectively. These results show that refrigerated storage and UV irradiation of table grapes can be beneficial in terms of increasing the content of potentially health-promoting phenolics. PMID- 11052708 TI - Behavior of monosaccharides, phenolic compounds, and color of red wines aged in used oak barrels and in the bottle. AB - A red wine with appropriate basic quality characteristics for aging was stored in oak barrels for 12 months and then bottled and aged for a further 6 months. The same ambient conditions of temperature and humidity were maintained throughout the entire aging process. The barrels used were made from three different species of oak by four different cooperages and had been used for at least two years. Analysis of variance and principal component analysis were run on the values for hexoses, pentoses, total anthocyanins, ortho-diphenols, low- and high-polymer polyphenols, and color parameters to study the behavior of the monosaccharides and polyphenols in response to the factors of aging time, the oak variety employed, and the source cooperage where the barrels had been made. Time trends for all the phenolic components were directly related to aging time, with low polymeric polyphenols (LPPs) being the most affected by wood type and source cooperage. Wine color was defined by a basic red color which decreased with aging time in the barrel and was altered by yellowish pigment components differing for each of the barrels in which oxidative aging took place and by increased stability of the blue copigments. Principal component analysis showed that samples of the same source wine aged in different barrels tended to be grouped together according to each of the aging intervals considered. PMID- 11052709 TI - Fining treatments of white wines by means of polymeric adjuvants for their stabilization against browning. AB - Browning and maderization represent important problems for white wine stability. Essentially, this is due to polyphenol oxidation in the wine. The problem has been remedied by adsorption of polyphenol compounds with polymeric adjuvants (chitosans, scleroprotein, and polylactic acid) not used traditionally in wine making. In particular, some chitosans reduced the polyphenol content and stabilized two Italian white wines (Trebbiano and Albana) to the same extent as did potassium caseinate, an adjuvant normally used in enology. Moreover, chitosans could be reused after a simple regeneration process. PMID- 11052710 TI - Estrogenic effects of extracts from cabbage, fermented cabbage, and acidified brussels sprouts on growth and gene expression of estrogen-dependent human breast cancer (MCF-7) cells. AB - Cruciferous vegetable extracts from freeze-dried cabbage (FDC), freeze-dried fermented cabbage (FDS), and acidified Brussels sprouts (ABS) were prepared by exhaustive extraction with ethyl acetate. Estrogenic and antiestrogenic effects of these extracts were analyzed. To identify whether the extracts are potential estrogen receptor (ER) ligands that can act as agonists or antagonists, the binding affinity of extracts for the ER was measured using a competitive radiometric binding assay. The extracts bound with low affinity to the ER, and the relative binding affinity is estradiol > FDS > FDC > ABS. These extracts were evaluated for their estrogenic and antiestrogenic activities in estrogen dependent human breast cancer (MCF-7) cells using as endpoints proliferation and induction of estrogen-responsive pS2 gene expression, which was analyzed using Northern blot assay. At low concentrations (5-25 ng/mL) all of the extracts reduced 1 nM estradiol-induced MCF-7 cell proliferation. Extracts at 25 ng/mL also inhibited estradiol-induced pS2 mRNA expression. At higher extract concentrations (50 ng/mL-25 microg/mL), however, increased proliferation in MCF-7 cells was observed. Similarly, expression of the pS2 gene was induced by higher extract concentrations (0.25-25 microg/mL). The pure estrogen antagonist, ICI 182,780, suppressed the cell proliferation induced by the extracts as well as by estradiol and also the induction of pS2 expression by the extracts. The ER subtype-selective activities of FDC and FDS were analyzed using a transfection assay in human endometrial adenocarcinoma (HEC-1) cells. FDS acted as an ERalpha selective agonist while FDC fully activated both ER-alpha and ER-beta. Growth of the ER-negative MDA-231 cells was not affected by the extracts or by estradiol. This study demonstrates that cruciferous vegetable extracts act bifunctionally, like an antiestrogen at low concentrations and an estrogen agonist at high concentrations. PMID- 11052711 TI - Autoxidation of packed almonds as affected by maillard reaction volatile compounds derived from roasting. AB - Shelled almonds of two Italian varieties, Romana and Pizzuta, peeled and unpeeled, were roasted and packed under different conditions: air (control), vacuum, and Maillard reaction volatile compounds (MRVc) derived from the roasting process. Samples were stored for approximately 8 months at room temperature, without light, and, at regular intervals, were collected and analyzed to evaluate the progress of lipid oxidation. Peroxide values, triglyceride oligopolymers, and oxidized triglycerides were evaluated during the storage time. Results showed that, although the MRVc atmosphere did not protect the lipid fraction of almonds as well as the vacuum condition; nevertheless, it was more protective than the control atmosphere, showing an antioxidant effect. The effect of the natural coating was a strong protection against lipid oxidation; in fact, only the unpeeled samples showed peroxide values lower than the threshold of acceptability (25 milliequiv of O(2)/kg of oil). Moreover, at the end of the storage period, Pizzuta almonds showed a greater deterioration than those of the Romana variety. PMID- 11052712 TI - Biological response of rats fed with tofu treated with high hydrostatic pressure. AB - Emerging technologies for food preservation have arisen in recent years, such as high-pressure (HP) hydrostatic treatment, and the biological response for this kind of food preservation is not well-known. Forty female rats (six weeks old) were used in the experiment to evaluate the biological effects of HP treatment of tofu. The animals were divided into groups that were fed with tofu (untreated), tofu treated with HP, and conventional food (as control) for 28 days. The glucose level, mineral content (calcium, potassium, zinc, and magnesium), shinbone maximum shear force, weight of the body, and weight of organs (heart, liver, spleen, and kidneys) were analyzed. The biological response for the rats was that significant differences were found in the calcium amount determined on the serum of the rats fed with untreated tofu and those fed with tofu treated with HP, and the calcium amount was lower on the rats fed with tofu treated with HP. Also, there were significant differences in the weight of the liver, and it was lower in the rats fed with tofu treated with HP. It was quite remarkable how the weight of the body and organs were smaller in the rats fed with tofu in comparison to the weight of the control rats. In the other components assayed no significant differences were found. HP produces a potential effect on tofu as it is observed in the rats response to the tofu treated with HP. PMID- 11052713 TI - Optimal conditions for phytate degradation, estimation of phytase activity, and localization of phytate in barley (cv. Blenheim). AB - Using a multivariate experimental design, optimal conditions for phytate degradation were found to be pH 4.8 and 57 degrees C in barley flour (cv. Blenheim) and pH 5.2 and 47 degrees C in a crude extracted phytase from barley. Three methods for measuring phytase activity in raw and hydrothermally processed barley were compared. Incubation at pH 5 and 55 degrees C for 60 min did not give significantly different results (p > 0.05), whereas incubation at pH 5 and 50 degrees C for 10, 20, 30, and 60 min gave significantly different results (p < 0.001) between methods. The change in microstructure of phytate globoids during hydrothermal processing showed that the degradation was highest in the scutellum cells and less in the aleurone layer. PMID- 11052714 TI - Novel procedure for identification of compounds inhibitory to transcription of genes involved in mycotoxin biosynthesis. AB - A novel assay is described for the identification and isolation of compounds that inhibit the transcription of genes involved in mycotoxin biosynthesis. The thin layer chromatography-based assay was used to screen plant extracts for compounds that would inhibit the expression of the beta-glucuronidase reporter gene under the control of an aflatoxin biosynthesis gene promoter in Aspergillus parasiticus. The assay was used to track purification of an inhibitory compound, cp2, from extracts of black pepper (Piper nigrum). Cp2 did not inhibit mycelial growth or the expression of the beta-tubulin gene but did inhibit aflatoxin biosynthesis at the transcriptional level. Applications of cp2 to the control of mycotoxins are discussed. PMID- 11052715 TI - Elucidation of fipronil photodegradation pathways. AB - The phenylpyrazole insecticide fipronil (I) photolyzes to its desthio product (II) in aqueous solution. However, the necessity of an intervening oxidation to a sulfone intermediate (III) has not been resolved, and the photodegradation products of II have not been identified. Using GC-MS, HPLC-UV/vis, electrospray MS, (19)F NMR, and GC-TSD, our objective was to characterize the photodegradation pathways of I, which would clarify the role of III, identify products of II, and explain unbalanced mass accounts in previous studies. Findings showed that II is formed directly and photochemically from I, confirmed by the greater stability of III (t(1/2) 112 h), and that successive oxidations of I to III and then a sulfonate (IV) comprise a second pathway. Compound II underwent photodechlorination, substitution of chlorine by trifluoromethyl, and pyrazole ring cleavage. This work is significant to understanding the photochemistry of novel phenylpyrazole pesticides in the environment. PMID- 11052716 TI - Reduction of pesticide residues on produce by rinsing. AB - In 1997 this laboratory initiated a research program with the objective of examining the effect that rinsing of produce with tap water would have on pesticide residues. Samples were obtained from local markets and/or grown at our experimental farm. Because approximately 35% of produce from retail sources contains pesticide residues, growing and treating produce at an experimental farm had the advantage that all such samples contain pesticide residues. Pesticides were applied under normal field conditions to a variety of food crops and the vegetation was allowed to undergo natural weathering prior to harvest. The resulting samples contained field-incurred or "field-fortified" residues. This experimental design was employed to mimic as closely as possible real world samples. Crops were treated, harvested, and divided into equal subsamples. One subsample was processed unwashed, whereas the other was rinsed under tap water. The extraction and analysis method used was a multi-residue method developed in our laboratory. Twelve pesticides were included in this study: the fungicides captan, chlorothalonil, iprodione, and vinclozolin; and the insecticides endosulfan, permethrin, methoxychlor, malathion, diazinon, chlorpyrifos, bifenthrin, and DDE (a soil metabolite of DDT). Statistical analysis of the data using the Wilcoxon signed-rank test showed that rinsing removed residues for nine of the twelve pesticides studied. Residues of vinclozolin, bifenthrin, and chlorpyrifos were not reduced. The rinsability of a pesticide is not correlated with its water solubility. PMID- 11052717 TI - Simultaneous derivatization and trapping of volatile products from aqueous photolysis of thiamethoxam insecticide. AB - An aqueous photolysis study was conducted with radiolabeled thiamethoxam, 4H 1,3,5-oxadiazin-2-imine, 3-[(2-chloro-5-thiazolyl)methyl]tetrahydro-5-methyl-N nitro, to establish the relevance of aqueous photolysis as a transformation process for (14)C-[thiazolyl]-thiamethoxam. (14)C-[thiazolyl]-thiamethoxam was applied to sterile sodium acetate pH 5 buffer solution at a dose rate of approximately 10 ppm. The resulting samples were incubated for up to 30 days at 25 degrees C under irradiated and nonirradiated conditions. The irradiated samples were exposed to a 12-hour-on and 12-hour-off light cycle. Volatile fractions accounted for up to an average of 56.76% of the total dose for the irradiated incubations and <0.08% for the nonirradiated incubations. These fractions were proposed to be a mixture of carbonyl sulfide (COS) and isocyanic acid (CONH). Verification of these components was accomplished by trapping with cyclohexylamine and formation of the thiocarbamate and the isocyanic acid derivatives. A similar method of trapping thiocarbamate metabolites was reported (Chen and Casida, 1978) where filter paper saturated with isobutylamine in methanol was arranged to trap (14)COS and (14)CO(2) under a positive flow of O(2) at 25 degrees C. Mass spectroscopy of the derivatized components confirmed the presence of carbonyl sulfide as the cyclohexylamine thiocarbamate and of isocyanic acid as its cyclohexylamine derivative. Evidence from this study indicates that thiamethoxam degrades significantly under photolytic conditions. PMID- 11052718 TI - In vitro and in situ inhibition of carotenoid biosynthesis in Capsicum annuum by bleaching herbicides. AB - Pepper leaves treated with the herbicide J852 show an accumulation of phytoene and zeta-carotene, whereas treatment with norflurazon led to an accumulation of only phytoene. The effects of these herbicides were examined in vitro after the expression of carotenoid desaturases in Escherichia coli. Whereas norflurazon is a potent inhibitor of phytoene desaturase (PDS) (I(50) = 0.12 microM) but not of zeta-carotene desaturase (ZDS) (I(50) = 144 microM), J852 inhibits both PDS (I(50) = 23 microM) and ZDS (I(50) = 49 microM). The influence of PDS/ZDS inhibition on gene expression was examined by comparative RT-PCR. None of the examined genes, namely, encoding phytoene synthase, PDS, ZDS, or the terminal oxidase associated with phytoene desaturation, were induced upon herbicide treatment in pepper leaves or seedlings. This was unexpected because inhibition of carotene desaturation led to an up-regulation of the carotenoid biosynthetic capacity (higher amounts of accumulating precursors plus remaining colored carotenoids are present in treated tissues versus control). PMID- 11052719 TI - Influence of metam sodium on the dissipation and residual biological activity of the herbicides EPTC and pebulate in surface soil under black plastic mulch. AB - Metam sodium is a potential replacement for methyl bromide, which is used to control soil pests. Metam sodium rapidly breaks down in the soil to form methylisothiocyanate (MITC). Dissipation of the herbicides EPTC and pebulate in a silt loam soil under plastic mulch in the absence and presence of metam sodium was examined in field experiments in 1998 and 1999 at Knoxville, Tennessee. EPTC half-life (DT(50)) was 9 d, but when applied in conjunction with metam sodium DT(50) increased to 22 d. Similarly, average pebulate DT(50) was 8 d and increased to 23 d when applied in conjunction with metam sodium. This increase in herbicide DT(50) with the addition of metam sodium is thought to be due to a reduction in soil microorganisms that degrade EPTC and pebulate. EPTC applied with metam sodium injured tomato plants and reduced total crop yield more than EPTC, pebulate, or pebulate with metam sodium. The increased tomato injury may have been related to the greater and prolonged activity of EPTC and slower EPTC dissipation in the presence of metam sodium or MITC. PMID- 11052720 TI - Isolation and characterization of fungal inhibitors from Epichloe festucae. AB - A series of studies was conducted to test the antifungal activity of clavicipitaceous endophytes and to identify potential fungal inhibitors in this symbiotic infection. A diverse group of endophytes was screened for antifungal activity using organic extracts from liquid fermentation cultures. Fungal inhibitors were purified from fermentation cultures of Epichloe festucae using a bioassay-directed extraction with Cryphonectria parasitica as the test organism. Compounds shown to have antifungal activity were subsequently identified using NMR and GC-MS. Extracts from a wide range of fungal isolates had various degrees of antifungal activity, but the greatest antifungal activity was observed in E. festucae and Neotyphodium tembladerae. Three types of inhibitors were isolated from a batch culture of E. festucae, including several indole derivatives, a sesquiterpene, and a diacetamide. Among the indole derivatives, indole-3-acetic acid and indole-3-ethanol were identified as the major indoles. These compounds were previously reported in endophytic fungi, and this study suggests a role in host disease resistance against other pathogens. The diversity in fungal inhibitors produced by this endophyte also suggests that fungal inhibitors may act additively or synergistically to reduce colonization of endophyte-infected hosts by potential fungal competitors. PMID- 11052721 TI - Identification of metabolites of [1,2,3-(13)C]Propargyl alcohol in mouse urine by (13)C NMR and mass spectrometry and comparison to rat. AB - Species differences in the metabolism of acetylenic compounds commonly used in the formulation of pharmaceuticals and pesticides have not been investigated. To better understand the in vivo reactivity of this bond, the metabolism of propargyl alcohol (PA), 2-propyn-1-ol, was examined in rats and mice. An earlier study (Banijamali, A. R.; Xu, Y.; Strunk, R. J.; Gay, M. H.; Ellis, M. C.; Putterman, G. J. J. Agric. Food Chem. 1999, 47, 1717-1729) in rats revealed that PA undergoes extensive metabolism primarily via glutathione conjugation. The current research describes the metabolism of PA in CD-1 mice and compares results for the mice to those obtained for rats. [1,2,3-(13)C;2,3-(14)C]PA was administered orally to the mice. Approximately 60% of the dose was excreted in urine by 96 h. Metabolites were identified, directly, in whole urine by 1- and 2 D (13)C NMR and HPLC/MS and by comparison with the available reference compounds. The proposed metabolic pathway involves glucuronide conjugation of PA to form 2 propyn-1-ol-glucuronide as well as oxidation of PA to the proposed intermediate 2 propynal. The aldehyde undergoes conjugation with glutathione followed by further metabolism to yield as final products 3,3-bis[(2-acetylamino-2-carboxyethyl)thio] 1-propanol, 3-[(2-acetylamino-2-carboxyethyl)thio]-3-[(2-amino-2-carboxyethyl)thi o]-1-propanol, 3,3-bis[(2-amino-2-carboxyethyl)thio]-1-propanol, 3-[(2-amino-2 carboxyethyl)thio]-2-propenoic acid, and 3-[(2-formylamino-2-carboxyethyl)thio]-2 propenoic acid. A small portion of 2-propynal is also oxidized to result in the excretion of 2-propynoic acid. On the basis of urinary metabolite data, qualitative and quantitative differences are noted between rats and mice in the formation of the glucuronide conjugate of PA and in the formation of 2-propynoic acid and metabolites derived from glutathione. These metabolites represent further variation on glutathione metabolism following its addition to the carbon carbon triple bond compared to those described for the rat. PMID- 11052722 TI - Effects of pH on chemical stability and de-esterification of fenoxaprop-ethyl by purified enzymes, bacterial extracts, and soils. AB - De-esterification is an initial step in the metabolism of certain herbicides, for example, fenoxaprop-ethyl [(+/-)-ethyl 2-[4-[(6-chloro-2 benzoxaolyl)oxy]phenoxy]propanoate] (FE). The ethyl-ester bond cleavage of FE to fenoxaprop acid (FA) by purified enzymes, crude bacterial enzyme preparations, and soils was investigated. In similar experiments fluorescein diacetate (FDA) was used as an alternative substrate. FE stability was pH sensitive in acidic buffered solutions; that is, below pH 4.6, rapid nonenzymatic hydrolysis of the benzoxazolyl-oxy-phenoxy ether linkage occurred, forming 6-chloro-2,3-dihydro benzoxazol-2-one (CDHB) and ethyl 4-hydroxyphenoxypropanoate or 4 hydroxyphenoxypropanoate. With porcine esterase and cell-free Pseudomonas fluorescens extracts, activity on FE and FDA was most rapid at pH 7.6-8.6 but decreased 80-90% at pH 5.6. Yeast (Candida cylindrica) lipase-mediated de esterification of FE and FDA was not as sensitive to pH; that is, activity at pH 4.6 was 70% of that at pH 7.6. Short-term incubations (20 h) were conducted in eight soils (pH 4.5-6.9) treated with (14)C-chlorophenyl ring-labeled FE (2 mg kg(-)(1)). In the most acidic soils (pH 4.4-4.5) 25% of the (14)C was recovered as FA, versus 30-40% in moderately acid soils (pH 5.0-5.6) and 55% in neutral soils (pH 6.8-6.9). There was a similar correlation between soil pH and FDA de esterification. CDHB was formed in all acidic soils with levels 4-fold greater in pH 4.4-4.5 soils than in pH 5. 0-5.6 soils. CDHB was not formed in neutral soils. Results demonstrate some chemical hydrolysis (benzoxazolyl-oxy-phenoxy ether linkage) of FE in acid soils, the sensitivity of enzymatic de-esterification of FE to pH, and the potential of FDA as a colorimetric indicator for esterase hydrolysis of FE. PMID- 11052723 TI - Comparative toxicity of selected organophosphate insecticides against resistant and susceptible clones of the greenbug, Schizaphis graminum (Homoptera: aphididae). AB - Comparative toxicity of selected organophosphate (OP) insecticides against resistant and susceptible clones of the greenbug, Schizaphis graminum, were studied both in vitro and in vivo. Two resistant (OR-1 and OR-2) clones of the greenbug showed marginal to high levels of resistance to all seven OPs tested, ranging from 11- to 327-fold greater than those of a susceptible (OSS) clone. The OR-1 clone showed lower levels of resistance to phenyl (parathion and parathion methyl) and heterocyclic (chlorpyrifos) OPs than to aliphatic OPs (dimethoate, omethoate, disulfoton, and demeton-S-methyl), whereas the OR-2 clone showed a rather broad spectrum of resistance to nearly all OP insecticides examined. In vitro inhibition of acetylcholinesterase (AChE) using six selected OP oxon analogues showed that alterations of AChE were involved in resistance to all OP compounds examined in both the OR-1 and OR-2 clones. Although the levels of insensitivity of AChE to these OPs were relatively low, ranging from 1.1- to 3.8 fold, the insensitivity spectrum of AChE to different OPs was rather broad. The general esterase activity in the OR-1 and OR-2 clones was 1.3-8. 4-fold higher than that in the OSS clone, depending on the substrates used. The AChE activity in both the OR-1 and OR-2 clones was 1.8-fold higher than that in the OSS clone. High resistance levels of the OR-2 clone to phenyl and heterocyclic OPs appeared to be associated with the ability of the esterases to hydrolyze beta-naphthyl acetate and more hydrophobic substrates. PMID- 11052724 TI - Antioxidative activity and carotenoid and tomatine contents in different typologies of fresh consumption tomatoes. AB - The phytonutrient intake associated with tomato consumption depends also on cultivar and fruit ripening stage. This work associates the antioxidative ability, the level of carotenoids, and the amount of glycoalkaloids to the main carpometric characteristics of four different typologies of tomatoes: "cherry", "cluster", "elongated," and "salad". These typologies have different weights and shapes, and they are usually consumed in the Mediterranean area at different ripening stages. Results showed that the considered tomato typologies also differ in their antioxidative ability and their carotenoid and glycoalkaloid contents. Growing conditions are also important in determining fruit characteristics: the analysis of the same cultivar of cherry tomato produced under the influence of moderate salt stress showed increases in the lipophilic antioxidative ability and the amount of carotenoid, whereas the level of glycoalkaloid decreased. PMID- 11052725 TI - Phlorin screening in various citrus species and varieties. AB - Phlorin (3,5-dihydroxyphenyl beta-D-glucopyranoside), an orange peel marker, has been searched in 45 species and varieties of Citrus. The phlorin content was determined by high-pressure liquid chromatography in juices and aqueous peel extracts of these different fruits. The phlorin content in C. reticulata peel extract varies from 0 to 1012 mg L(-)(1) with a mean of 162 mg L(-)(1). On the contrary, phlorin was not found in mandarin and clementine juices except for mandarin "Commune" and "Beauty" (33 and 30 mg L(-)(1), respectively). In the 14 species of oranges and varieties, phlorin was detected in juices and peel extracts with a mean of 22 and 492 mg L(-)(1), respectively, while for grapefruits, means were 108 mg L(-)(1) in juices and 982 mg L(-)(1) for peel extracts. Tangors and tangelos which are hybrids (C. reticulata x C. sinensis and C. reticulata x C. paradisi, respectively) are characterized by the systematic presence of phlorin in peels (mean: 406 and 659 mg L(-)(1), respectively) while in juices its presence could be variable (0-73 mg L(-)(1)). These heterogeneity and values may be explained by the genetic variability of these hybrids and the phlorin content of their parentage group. PMID- 11052726 TI - Identification by HPLC-DAD and HPLC-MS analyses and quantification of constituents of fennel teas and decoctions. AB - Qualitative and quantitative differences among the constituents in various fennel (Foeniculum vulgare Mill., family Apiaceae) teas prepared by classical infusion, microwave decoction, and dissolution are reported. Different commercial starting materials, such as fruit (unbroken and crushed), four herbal teas, and two instant herbal teas were evaluated. Chlorogenic acid (1), quercetin-3-O-beta-D glucuronide (2), p-anisaldehyde (3), and trans-anethole (4) were identified by HPLC-DAD and HPLC-MS as constituents of fennel teas. No coumarins, which are characteristic constituents of plants of Apiaceae family, were found. Trans anethole (4), the main constituent of the essential oil, was present in all teas. In addition p-anisaldehyde (3), a degradation product of trans-anethole, was also identified in all teas with the exception of two samples. Chlorogenic acid (1) and quercetin-3-O-beta-D-glucuronide (2) were also present in all teas. In addition, minor unidentified flavonol constituents were found in two teas. Quality, activity, and safety of the content of the investigated preparations are also discussed. PMID- 11052727 TI - Long-chain aliphatic beta-diketones from epicuticular wax of Vanilla bean species. Synthesis of nervonoylacetone. AB - Analysis of the neutral lipids from Vanilla fragrans and Vanilla tahitensis (Orchidaceae) without saponification resulted in the isolation and identification of a new product family in this plant: beta-dicarbonyl compounds. The compound structures are composed of a long aliphatic chain with 2,4-dicarbonyl carbons and a cis double bond at the n-9 position. They represent approximately 28% of the neutral lipids, that is, 1.5%, in immature beans, and approximately 10% of the neutral lipids, that is, 0.9%, in mature beans. Using retention indices, gas chromatography-mass spectrometry, derivatization reactions, and chemical degradation, five beta-dicarbonyl compounds have been identified including 16 pentacosene-2,4-dione, 18-heptacosene-2,4-dione, 20-nonacosene-2, 4-dione, 22 hentriacontene-2,4-dione, and 24-tritriacontene-2, 4-dione. Among them (Z)-18 heptacosene-2,4-dione, or nervonoylacetone, has been synthesized in two steps starting from nervonic acid. The major constituent, nervonoylacetone, represented 74.5% of the beta-dicarbonyl fraction. The range of these compounds has been studied in relation with bean maturity for V. fragrans and V. tahitensis species. This compound family has not been found in the leaves or stems of any of the three vanilla species studied and is markedly absent in the beans of V. madagascariensis. PMID- 11052728 TI - Flavonoids in monospecific eucalyptus honeys from Australia. AB - The HPLC analyses of Australian unifloral Eucalyptus honeys have shown that the flavonoids myricetin (3,5,7,3',4', 5'-hexahydroxyflavone), tricetin (5,7,3',4',5' pentahydroxyflavone), quercetin (3,5,7,3',4'-pentahydroxyflavone), luteolin (5,7,3', 4'-tetrahydroxyflavone), and kaempferol (3,5,7, 4'-tetrahydroxyflavone) are present in all samples. These compounds were previously suggested as floral markers of European Eucalyptus honeys. The present results confirm the use of flavonoid analysis as an objective method for the botanical origin determination of eucalyptus honey. Honeys from E. camaldulensis (river red gum honey) contain tricetin as the main flavonoid marker, whereas in honeys from E. pilligaensis (mallee honey), luteolin is the main flavonoid marker, suggesting that species specific differences can be detected with this analysis. The main difference between the flavonoid profiles of Australian and European Eucalyptus honeys is that in the Australian honeys, the propolis-derived flavonoids (pinobanksin (3,5, 7-trihydroxyflavanone), pinocembrin (5,7-dihydroxyflavanone), and chrysin (5,7 dihydroxyflavone)) are seldom found and in much smaller amounts. PMID- 11052729 TI - Foliar application of selenite and selenate to potato (Solanum tuberosum): effect of a ligand agent on selenium content of tubers. AB - The effect of a foliar spray of selenium on potatoes was investigated for 2 years. Amounts of 0, 50, and 150 g of Se ha(-)(1) were applied both as sodium selenate and as sodium selenite in water, either pure or with the addition of 0.15% of soluble leonardite as a source of humic acids (pH 7). Tuber selenium concentration increased with the application levels, both with sodium selenate and with sodium selenite, when only aqueous solutions were used. When humic acids were added, the tuber selenium level rose more markedly after the application of sodium selenate as compared to the case of the aqueous solutions; however, in the case of sodium selenite, the level showed a large increase only after the application of 50 g of Se ha(-)(1). Kinetics showed that humic acids raised the selenate availability, but no differences were found in the distribution of selenium in the tuber fractions. Foliar application of selenium with humic acids was proven to be a good way to increase the selenium content of potatoes, but the assimilation process of selenium was simpler with selenate than with selenite. PMID- 11052730 TI - Behavior of four sulfonylurea herbicides in the presence of hydroxy compounds. AB - The behavior of four sulfonylurea herbicides (metsulfuron methyl, chlorsulfuron, chlorimuron ethyl, and bensulfuron methyl) was studied in the presence of various hydroxy compounds. When dissolved at 30 degrees C in simple primary, secondary, or tertiary alcohols (methanol, ethanol, isopropyl alcohol, and tert-butyl alcohol) and in glycerol or in poly(ethylene glycol), most of these herbicides underwent rapid alcoholysis involving the breakdown of the urea part of the molecule. The corresponding sulfonyl carbamate is recovered in high yields, along with a small amount of sulfonylamide formed in the concomitant hydrolysis. Degradation rate constants and the selectivity of conversion were established. The addition of buffered water (pH 7.0) inhibited the alcoholysis reaction, leaving only hydrolysis, as already observed with concentrated saccharide solutions. In phenol solution, slight herbicide hydrolysis was primarily observed. It appeared that alcoholysis reactions only occurred under very particular conditions when sulfonylurea herbicides are dissolved in pure alcohols, without buffered water. These results led to the conclusion that in soil, similar alcoholysis reactions seem unlikely. PMID- 11052731 TI - Effect of temperature and moisture on the degradation and sorption of florasulam and 5-hydroxyflorasulam in soil. AB - The degradation rate and sorption characteristics of the triazolopyrimidine sulfonanilide herbicide florasulam and its principal degradation product 5 hydroxyflorasulam (5-OH-florasulam) were determined as a function of temperature and moisture in three different soils. The half-life for degradation of florasulam ranged from 1.0 to 8.5 days at 20-25 degrees C and from 6.4 to 85 days at 5 degrees C. The half-life for degradation of 5-OH-florasulam ranged from 8 to 36 days at 20-25 degrees C and from 43 to 78 days at 5 degrees C. The degradation rate of both compounds was strongly influenced by temperature, with activation energies ranging from 57 to 95 kJ/mol for florasulam and from 27 to 74 kJ/mol for 5-OH florasulam. Soil moisture content had negligible impact on the degradation rate. Apparent (nonequilibrium) sorption coefficients for florasulam and 5-OH florasulam at 0 days after treatment (DAT) were 0.1-0.6 L/kg and increased linearly with time for both florasulam and 5-OH-florasulam (r(2) > 0.90) to levels as high as 12-23 L/kg. Heats of adsorption were calculated on one soil as a function of time. Heat of adsorption values for both florasulam and 5-OH florasulam increased as incubation time increased and the amount of each compound decreased; values were near 0 kJ/mol initially and increased to a maximum of 91 and 66 kJ/mol for florasulam and 5-OH-florasulam, respectively. PMID- 11052732 TI - Organo-clay formulations of the hydrophobic herbicide norflurazon yield reduced leaching. AB - The study aimed to reduce leaching of the hydrophobic herbicide norflurazon (4 chloro-5-methylamino-2-(alpha,alpha, alpha)-trifluoro-m-tolylpyridazin-3-(2H) one) by adsorbing it on clays or organo-clays. The surface of the clay mineral montmorillonite was modified from hydrophilic to hydrophobic by preadsorbing it with organic cations, of which thioflavin-T (TFT) at a loading corresponding to (5)/(8) of the cation-exchange capacity of the clay mineral yielded the highest affinity of adsorption of norflurazon. Pillared clay (PC) used without organic cations exhibited enhanced affinity for norflurazon adsorption, much higher than that of montmorillonite or sepiolite. Fourier transform infrared (FTIR) results showed interactions between aromatic moieties of preadsorbed TFT and the herbicide. Stronger interaction of the herbicide with a clay mineral or organo clay corresponded to its slower release. Formulations prepared on the basis of montmorillonite-TFT and PC were more effective in reducing herbicide leaching in soil columns in comparison to the commercial formulation, whereas the herbicidal efficiencies were comparable. PMID- 11052734 TI - Adsorption-desorption studies of selected herbicides in soil-Fly ash mixtures. AB - Fly ash and soil mixtures with a range of fly ash content from 0 to 100% were used to study the adsorption and desorption of herbicides atrazine, propazine, prometryne, propanil, and molinate in batch experiments. The isotherms shapes according to Giles classification (Giles et al., 1960) were S, L, and H as the substrate changed from sandy clay loam (SCL) to fly ash, depending on the percent of fly ash in the mixture. The adsorption isotherms fit the Freundlich equation x/m = K(f) C(1/)(n)(). The K(f) values increase with the increase of the fly ash content. The mean percent amounts of herbicides, for a range of concentration 1 20 mg L(-)(1), adsorbed on the soil were 21.9% for atrazine, 50.7% for propazine, 29.04% for prometryne, 43.14% for molinate, 31.35% for propachlor, and 46.34% for propanil. Mass balance estimations show that the adsorbed amounts of the herbicides increase along with the fly ash content in the sorbent mixture and reach the 99% in the "pure" fly ash. In contrast, the amounts desorbed with water decrease as the fly ash content increases. The n values ranged from 0.82 to 3.05 indicating that the carbon content of fly ash plays a significant role during the sorption process and an increase of heterogenity of solid substrate. The increase of the amounts desorbed with acetone indicates that the sorption of organic compounds onto fly ash is believed to occur principally via the weak induction forces of London or dispersion forces which are characteristic of the physical adsorption process. The results of this research demonstrate that the fly ash shows a significant capacity for adsorption of organic compounds from aqueous solution. PMID- 11052733 TI - Photostabilization of the herbicide norflurazon by using organoclays. AB - Photostable formulations of the herbicide norflurazon [4-chloro-5-(methylamino)-2 (alpha,alpha, alpha-trifluoro-m-tolyl)pyridazin-3-(2H)-one] were achieved by adsorbing it on pillared clay or on montmorillonite preadsorbed with the organic cation thioflavin T (TFT). Diffuse reflectance Fourier transform infrared spectra showed the existence of strong interactions between the aromatic moieties of preadsorbed TFT and the herbicide, particularly after irradiation. The photostabilization of norflurazon obtained with TFT-clay was mainly due to energy transfer from the herbicide to the organic cation via pi-pi interactions. An additional mechanism is the lower production of radicals from the clay when the clay mineral surface is covered with the organic cation. These radicals are responsible for the enhanced photodegradation observed when norflurazon was irradiated in the presence of untreated montmorillonite. PMID- 11052735 TI - Montmorillonite-phenyltrimethylammonium yields environmentally improved formulations of hydrophobic herbicides. AB - This study aimed to design formulations of hydrophobic herbicides, alachlor and metolachlor, by adsorbing them on the clay mineral montmorillonite preadsorbed by the small organic cation phenyltrimethylammonium (PTMA). An adsorption model that considers electrostatics and specific binding and the possibility of cation adsorption above the cation exchange capacity (CEC) could explain and yield predictions for PTMA adsorption in the presence of NaCl concentrations from 0 to 500 mM. Adsorption of alachlor and metolachlor from aqueous solution on a clay mineral preadsorbed by PTMA was determined by GC and modeled by Langmuir equation. Herbicide interactions with the organoclay were studied by Fourier transform infrared spectroscopy. Leaching of herbicides was determined by a bioassay using a column technique and Setaria viridis as a test plant. The adsorbed amounts of alachlor and metolachlor on montmorillonite preadsorbed by PTMA at a loading of 0. 5 mol/kg (Mont-PTMA0.5) were higher than at a loading up to the CEC, that is, 0.8 mol/kg, and were higher than those obtained by using several other organic cations. Herbicide formulations based on Mont-PTMA0.5 yielded the largest shifts of the infrared peaks of the herbicides. These formulations based on Mont-PTMA0.5 gave slower release and showed improved weed control in comparison with formulations based on other organoclays. These formulations maintained herbicidal activity in the topsoil and yielded the most significant reduction in herbicide leaching. PMID- 11052736 TI - Aroma potential of two bairrada white grape varieties: Maria Gomes and Bical. AB - Maria Gomes and Bical are the main white grape varieties in Portuguese Bairrada Appellation, which represent 80% and 15%, respectively, of white vineyard. To estimate their aroma potentialities, free and potential volatile components from the musts were examined. The free volatile components were extracted using a liquid-liquid continuous method and were analyzed by gas chromatography-mass spectrometry. The potential volatile compounds were determined after elimination of the free components by heat and enzymatic treatments. Principal component analysis was used to establish relations between the compounds and the varieties and also the form (free or in potential). Maria Gomes has 11.5 mg/L volatile compounds, of which 33% are in free form; Bical has 4.4 mg/L volatile compounds, of which 46% are in free form. A total of 59 compounds was identified and quantified. In Maria Gomes, the sum of the terpenoids is within the perception limits for hotrienol (0.21 mg/L) and linalool (0.20 mg/L). In Bical, benzyl alcohol and phenylethylethanol represent 20% of the volatile compounds. Considering that the volatile composition pattern of Maria Gomes and Bical varieties are different, wine-making technologies should be developed specifically for each variety. PMID- 11052737 TI - Structured fluids as microreactors for flavor formation by the Maillard reaction. AB - Thermal reactions of cysteine/furfural and cysteine/ribose mixtures were studied in model systems to gain more insight into the influence of structured fluids such as L(2) microemulsions and cubic phases on the generation of aroma compounds. Formation of 2-furfurylthiol from cysteine/furfural was particularly efficient in L(2) microemulsions and cubic phases compared to aqueous systems. The reaction led to the formation of two new sulfur compounds, which were identified as 2-(2-furyl)thiazolidine and, tentatively, N-(2-mercaptovinyl)-2-(2 furyl)thiazolidine. Similarly, generation of 2-furfurylthiol and 2-methyl-3 furanthiol from cysteine/ribose mixtures was strongly enhanced in structured fluids. The cubic phase was shown to be even more efficient in flavor generation than the L(2) microemulsion. It was denoted "cubic catalyst" or "cubic selective microreactor". The obtained results are interpreted in terms of a surface and curvature control of the reactions defined by the structural properties of the formed surfactant associates. PMID- 11052738 TI - Determination of antioxidant properties of aroma extracts from various beans. AB - Aroma extracts from fresh soybeans, mung beans, kidney beans, and azuki beans were prepared using simultaneous steam distillation and solvent extraction (SDE) under mild conditions (55 degrees C and 95 mmHg). Extracts were examined for antioxidative activities in two different assays. The aroma extracts isolated from all beans inhibited the oxidation of hexanal for nearly one month at a level of 250 microL/mL. Mung bean and soybean extracts inhibited malonaldehyde (MA) formation from cod-liver oil by 86% and 88%, respectively, at the 250 microL/mL level. Azuki and kidney bean extracts inhibited MA formation from cod-liver oil by 76% and 53%, respectively, at the 250 microL/mL level. The antioxidative activities of mung bean and soybean extracts were comparable with that of the natural antioxidant, alpha-tocopherol (vitamin E). PMID- 11052739 TI - 9-Hydroxypiperitone beta-D-glucopyranoside and other polar constituents from dill (Anethum graveolens L.) herb. AB - A methanolic extract from dill (Anethum graveolens) herb was subjected to XAD-2 adsorption chromatography. The methanolic eluate was fractionated with the all liquid chromatographic technique of multilayer coil countercurrent chromatography (MLCCC). After acetylation of MLCCC subfractions and flash chromatography, final purification of dill herb constituents was achieved by preparative and/or analytical HPLC. Nine compounds were obtained in pure form, including the beta-D glucopyranosides of 9-hydroxypiperitone, p-menth-2-ene-1,6-diol, and 8 hydroxygeraniol. Structure elucidation is based on electrospray ionization ion trap multiple mass spectrometry (ESI-MS/MS) as well as one- and two-dimensional nuclear magnetic resonance spectroscopy. PMID- 11052740 TI - Biotransformation of (-)-dihydromyrcenyl acetate using the plant parasitic fungus Glomerella cingulata as a biocatalyst. AB - The microbial transformation of (-)-dihydromyrcenyl acetate was investigated using the plant parasitic fungus Glomerella cingulata. As a result, (-) dihydromyrcenyl acetate was converted to dihydromyrcenol, 3,7-dihydroxy-3,7 dimethyl-1-octene-7-carboxylate, 3,7-dihydroxy-3,7-dimethyl-1-octene, 3,7 dimethyloctane-1,2, 7-triol-7-carboxylate, and 3,7-dimethyloctane-1,2,7-triol. In addition, microbial transformation of dihydromyrcenol by G. cingulata was carried out. The metabolic pathway of (-)-dihydromyrcenyl acetate is discussed. PMID- 11052741 TI - Organoleptic impact of 2-methoxy-3-isobutylpyrazine on red bordeaux and loire wines. Effect of environmental conditions on concentrations in grapes during ripening. AB - The 2-methoxy-3-isobutylpyrazine content in grapes and red wines was assayed by stable isotope dilution gas chromatography-mass spectrometry, following vapor extraction and purification on a cation resin microcolumn. The threshold beyond which the green bell pepper character is marked in wines has been determined. From a comparison of the 2-methoxy-3-isobutylpyrazine concentrations of 50 red Bordeaux and Loire wines from different vintages and grape varieties (Cabernet Sauvignon, Cabernet franc, and Merlot) with the intensity of the green bell pepper character as perceived on tasting, the threshold value was estimated to be 15 ng/L. Statistical analysis of the 2-methoxy-3-isobutylpyrazine concentrations of 89 red Bordeaux wines showed that Cabernet wines were more commonly affected by this vegetative character. Changes in the 2-methoxy-3-isobutylpyrazine concentration as the grapes ripen are affected by the environmental and cultural conditions (soil, climate, training system, etc.). A very good correlation was shown between the breakdown of malic acid and 2-methoxy-3-isobutylpyrazine as the grapes ripened, irrespective of grape variety, type of soil, or weather conditions. PMID- 11052742 TI - Identification and quantification of geosmin, an earthy odorant contaminating wines. AB - Musty, earthy odors are highly detrimental to the aromatic quality of wines. A characteristic aroma of freshly tilled earth, damp cellar was studied in some red and white wines of different origins. The extraction and purification of the wines marked by this odor have shown after analysis by gas chromatography olfactometry a unique strong odorous zone having the same odor as the one perceived at tasting. The compound responsible for this odorous zone was identified by gas chromatography-mass spectrometry as geosmin (trans-1,10 dimethyl-trans-9-decalol), which possesses a distinctive earthy odor. Geosmin may be present in wines at levels higher than the racemic geosmin olfactory perception threshold, thus suggesting its contribution to their off-aroma. Moreover, the presence of this compound in juice taken from freshly crushed grapes suggests that microorganisms that develop on the grapes may contribute to the presence of this compound in wines. PMID- 11052743 TI - Development and characterization of a flavoring agent from oyster cooker effluent. AB - The general composition of concentrated oyster cooker effluent (OCE) was 80% moisture, 6.7% total nitrogen, 2.4% glycogen, and 8.5% ash. Optimum conditions for enzymatic hydrolysis of OCE were 50 degrees C, 2 h of reaction time, 0.1% amylase mixture (alpha-amylase plus glucoamylase), and 0.2% protease NP. Hydrolysis of OCE led to an increase in free amino acids, with taurine comprising approximately 20% of the total. Inosine monophosphate was predominant (456 mg/100 g) among nucleotides and related compounds. Enzyme hydrolysis increased extractable nitrogen by approximately 2-fold. Trimethylamine, trimethylamine oxide, and total creatinine levels were not affected by enzyme treatment. Predominant aroma-active components of enzyme-hydrolyzed OCE included 2-acetyl-1 pyrroline and 3-(methylthio)propanal. Results of this study may help alleviate the wastewater disposal problem currently caused by OCE. PMID- 11052744 TI - Volatile compounds in the production of liquid beet sugar. AB - Samples from different parts of a beet sugar factory and refinery were analyzed with respect to volatile compounds by means of liquid-liquid extraction followed by gas chromatography-mass spectrometry (GC-MS). A limited number of the samples were analyzed by means of gas phase extraction (headspace) followed by GC-MS. Selected compounds were followed through the sugar manufacturing process. The behavior of different compounds varied greatly throughout the process, with some compounds such as geosmin (trans-1, 10-dimethyl-trans-9-decalol), dimethyl disulfide, and propionic and hexanoic acid present at the beginning of the process but disappearing rapidly after further processing. Other compounds, such as indole, dihydrobenzofuran, and 2-phenylethanol, were not detected at the start of the process but were formed later on and removed in the final product. In the final product, three pyrazines remained at fairly low concentrations, together with 3-methylcyclopentadione, ethylhexanol, and methyl pyrrole ketone. PMID- 11052745 TI - Aroma of fresh oysters Crassostrea gigas: composition and aroma notes. AB - In contrast to many foods, very little is known about the aroma of fresh oysters. This study deals with the relationship between extracted volatiles of oysters and their olfactory properties. Nearly 50 volatiles were identified: most of them were principally related to fatty acid oxidation (86%) and particularly to n-3 polyunstaturated fatty acid oxidation (66%). Only one volatile arose from amino acid degradation. Panelists detected 42 odors by sniffing. Among them, only 12 odors were definitely attributed to identified volatile. These odors were green/sulfur/crustacean, mushroom/citrus, and marine/cucumber notes and were attributed to dimethyl sulfide, 1-penten-3-one, hexanal, (2,4)-E,E-heptadienal, 1 octen-3-one, 1-octen-3-ol, 6-methyl-5-hepten-3-one, octanal, (E,Z)-2,6 nonadienal, (E)-2-octenal, and decanal, respectively. PMID- 11052746 TI - Dynamic headspace gas chromatography/mass spectrometry characterization of volatiles produced in fish oil enriched mayonnaise during storage. AB - Protection against lipid oxidation and formation of unpleasant fishy and rancid off-flavors in oil-in-water food emulsions, such as fish oil enriched mayonnaise, is difficult to achieve. Volatile profiles from stored mayonnaises with different oil phase compositions were collected using a developed dynamic headspace sampling technique, in which interfering acetic acid was removed in situ with potassium hydroxide, and subsequently 148 volatiles were characterized and monitored by gas chromatography/mass spectrometry. Multivariate statistics showed correlation between the concentration of 62 volatiles and the fish oil and storage parameters, indicating the formation of lipid oxidation products, which impose fishy off-flavors. Further verification was obtained by gas chromatography/olfactometry, by which, among 78 odors, cis-4-heptenal and trans,cis-2,4-heptadienal were detected as distinct fishy notes. In total, 27 volatiles, including 1-penten-3-one, cis-2-penten-1-ol, cis-3-hexenal, cis-4 heptenal, 1-octen-3-one, 1,cis-5-octadien-3-one, 1-octen-3-ol, trans,cis-2, 4 heptadienal, and trans,cis-2,6-nonadienal, were suggested to contribute to the developed unpleasant fishy and rancid off-flavors. PMID- 11052747 TI - Composition of the essential oil of Citrus tamurana Hort. ex Tanaka (Hyuganatsu). AB - The composition of the essential oil of Citrus tamurana Hort. ex Tanaka (Hyuganatsu), isolated by the cold-pressing method, was investigated by capillary GC and GC-MS. The effects of harvesting time, degree of freshness, and size of fruits on the composition of Hyuganatsu peel oils were also determined. A total of 126 volatile constituents were confirmed in the Hyuganatsu oils. The Hyuganatsu oils contained hydrocarbons (95.95-96.95%), aldehydes (0.33-0.62%), alcohols (1.91%-2.64%), ketones (0.40-0.62%), esters (0.28-0.39%), oxides (0.04 0.06%), acids (0.01%), and trace amounts of fugenol methyl ether. Monoterpene hydrocarbons were predominant. Limonene (80.35-82.39%), gamma-terpinene (7.71 9.03%), myrcene (2.11-2.28%), linalol (1.37-2.01%), and alpha-pinene (1.17-1.43%) were the most abundant components in Hyuganatsu oils. The principal sesquiterpene hydrocarbon was trans-beta-farnesene (0.60-1.04%), and its content in Hyuganatsu oils was higher than in oils of other citrus fruits. The number of ketones and the content of l-carvone in Hyuganatsu oils were higher than in other citrus oils. PMID- 11052748 TI - Activities of antioxidants are affected by colloidal properties of oil-in-water and water-in-oil emulsions and bulk oils. AB - The activity of alpha-tocopherol, Trolox, propyl gallate, gallic acid, methyl carnosoate, and carnosic acid was studied in two oil-in-water (o/w) emulsions, in two water-in-oil (w/o) emulsions, and in bulk oil with and without added emulsifiers. All antioxidants had either moderate or higher activity in bulk oil than in the emulsions. In most emulsions, the most polar antioxidants, propyl gallate and gallic acid, exhibited either prooxidant activity or no antioxidant activity. Methyl carnosoate was the most active antioxidant in w/o emulsions but was less active than Trolox in o/w emulsions. alpha-Tocopherol was less active in bulk oil than in emulsions, but its activity in bulk oil was markedly enhanced by the addition of o/w emulsifiers. Partitioning of antioxidants, hydrogen bonding, interphase transport, surface accessibility, and interaction of emulsifier with antioxidants are considered to be important parameters that determine antioxidant activity in lipid-containing systems. PMID- 11052749 TI - Simple rapid procedure for preparation of large quantities of ovalbumin. AB - A simple rapid procedure for preparation of large quantities of highly purified homogeneous ovalbumin from egg white by using an anion exchanger is described. It is based on the principle of frontal chromatography. The volume of "mucin-free" egg white loaded onto the column was determined in order to exceed resin capacity. Thus, competition between proteins for resin sites was created. Owing to its high negative charge density, ovalbumin drives other egg white proteins from the column progressively. Two hundred and fifty milliliters of Q-Sepharose FF gel eluted isocratically with 0. 5 M NaCl extracted 9.55 g of ovalbumin with a purity rate of 83%. A 6.75 g amount of ovalbumin, with a purity rate of 94%, was recovered with an isocratic elution program using 0.14 M NaCl. Purified ovalbumins were compared by electrophoresis and analytical chromatography with other ovalbumin preparations. PMID- 11052750 TI - Approaches to wine aroma: release of aroma compounds from reactions between cysteine and carbonyl compounds in wine. AB - Under conditions close to those of wine, that is, low pH, aqueous medium, and low temperatures, this work examines the role of carbonyl (acetoin and acetol) and dicarbonyl (glyoxal, methylglyoxal, diacetyl, and pentane-2,3-dione) compounds associated with cysteine in the formation of odorous products. In particular, thiazole, 4-methylthiazole, 2-acetylthiazole, and trimethyloxazole and two sulfur and oxygenated heterocyclic compounds, 2-furanmethanethiol and thiophene-2-thiol, are examined. For thiophene-2-thiol, the reactional mechanism is proposed. Attempts were made to detect these compounds in wines from various origins. Certain molecules were identified for the first time in wine. PMID- 11052751 TI - Effect of storage temperature on the degradation of dimethoate in fortified orange and peach juices. AB - The effect of storage temperature on dimethoate degradation in fortified orange and peach juices was studied. The insecticide was aseptically injected into packed orange and peach juices and stored at 40, 15, and 0 degrees C. Samples were taken at regular time intervals and were examined for dimethoate residues. The residues were determined with a simple gas chromatographic method; the recoveries of dimethoate from orange and peach juices were found to be from 88 to 114% for both products. The limits of determination were 0.004 and 0.003 mg/kg, respectively. From the experimental data, rate constants, half-lives, and activation energies for the decomposition of dimethoate in orange and peach juices were evaluated. During the storage of fruit juices in refrigerated rooms (0 degrees C) half-lives of dimethoate were found to be largely extended, being 1733 days for orange juice and 2310 days for peach juice. Corresponding times for storage at 15 degrees C were 533 days for both juices and for storage at 40 degrees C 24 days for orange juice and 24.6 days for peach juice. The activation energy for dimethoate in orange juice was 22.3 kcal/mol and for peach juice, 21. 2 kcal/mol. PMID- 11052752 TI - Tetrahydro-beta-carbolines, potential neuroactive alkaloids, in chocolate and cocoa. AB - Tetrahydro-beta-carbolines (THbetaCs), potential neuroactive alkaloids, were found in chocolate and cocoa. 6-Hydroxy-1-methyl-1,2, 3,4-tetrahydro-beta carboline (6OHMTHbetaC), 1,2,3, 4-tetrahydro-beta-carboline-3-carboxylic acid (THCA), 1-methyl-1,2,3, 4-tetrahydro-beta-carboline-3-carboxylic acid (MTCA) in both diastereoisomers (1S,3S and 1R,3S), and 1-methyl-1,2,3, 4-tetrahydro-beta carboline (MTHbetaC), besides serotonin and tryptamine biogenic amines, were identified and quantified in dark chocolate, milk chocolate, cocoa, and chocolate containing cereals by RP-HPLC-fluorescence and HPLC-MS. For each THbetaC, the concentration ranges were determined: 6OHMTHbetaC (0.16-3.92 microg/g), THCA (0.01-0.85 microg/g), 1S,3S-MTCA (0.35-2 microg/g), 1R,3S-MTCA (0.14-0.88 microg/g), and MTHbetaC (nd-0.21 microg/g). The highest content was generally found in chocolates and cocoas, but cereals containing chocolate also showed an appreciable amount of THbetaCs. The possible biological implications of this novel group of alkaloids in chocolate are discussed. PMID- 11052753 TI - Early structural changes in myosin rod upon heating of carp myofibrils. AB - Upon heating carp myofibrils at 40 degrees C, the amount of myosin that is soluble and monomeric dropped very quickly, roughly 5 times faster than the ATPase inactivation. This rapid decrease of solubility was well explained by a rapid denaturation of the rod portion as measured by chymotryptic digestibility. Chymotryptic digestion of heated myofibrils in a low-salt medium with EDTA generated a reduced amount of rod and subfragment-1 (S-1). The decrease of S-1 produced from the heated myofibrils was consistent with the ATPase inactivation. The decrease of rod produced from the heated myofibrils was explained by the increased susceptibility of the heavy meromyosin (HMM)/light meromyosin (LMM) junction to chymotryptic. It was, therefore, concluded that the fastest event occurring in the myosin molecule upon heating of myofibrils is the irreversible exposure of the HMM/LMM junction. PMID- 11052754 TI - Inhibition of formation of oxidative volatile components in fermented cucumbers by ascorbic acid and turmeric. AB - Two naturally occurring antioxidants, ascorbic acid and turmeric, were effective in inhibiting formation of hexanal, (E)-2-penenal, (E)-2-hexenal, (E)-2-heptenal, and (E)-2-octenal when slurries of fermented cucumber tissue were exposed to oxygen. Added ascorbic acid prevented formation of most of these oxidative aldehydes at 175 ppm or greater. Turmeric, which is used commercially as a yellow coloring in cucumber pickle products, was found to almost completely prevent aldehyde formation at 40 ppm. PMID- 11052755 TI - Tryptophan-N-glucoside in fruits and fruit juices. AB - In extracts prepared from various fruits as well as in fruit juices a single tryptophan glycoconjugate was detected by HPLC-MS analysis. Product ion spectra demonstrated the N-glycosidic linkage of a hexose moiety to the indole nitrogen. For structure elucidation, the novel tryptophan glycoside was isolated from pear juice and identified as N(1)-(beta-D-glucopyranosyl-(4)C(1))-L-tryptophan by (1)H, HH-COSY and (13)C NMR spectroscopy. Finally, we disclosed the biosynthetic origin of the novel tryptophan metabolite by demonstrating the enzymatic glycosylation of deuterium-labeled tryptophan, which was applied to pear fruit. PMID- 11052756 TI - Separation of thermostable pectinmethylesterase from marsh grapefruit pulp. AB - A rapid, ion-exchange method that isolates thermostable pectinmethylesterase (TS PME) from Marsh grapefruit pulp is presented. TS-PME was selectively extracted with 1 M NaCl and equilibrated at low pH (3.1 +/- 0.04). After dilution to a final concentration of 50 mM acetate buffer, pH 5.5, and 0.1, 0.25, or 0.5 M NaCl, the extract was applied to an ion-exchange column without ammonium sulfate precipitation and dialysis. The percent yield varied from an average of 70% (0.1 M NaCl) to 14% (0.5 M NaCl), with a specific activity of >400 U/mg protein in the latter sample. TS-PME activity of some column fractions varied compared with crude extracts before column separation. Some fractions lost thermostability after separation if loaded at 0.1 M or 0.25 M NaCl. In extracts that were loaded at 0.5 M NaCl, the TS-PME activity was significantly higher than that observed in the heated crude extracts. Ion-exchange chromatography may have separated an unidentified protective factor in PME fractions. PMID- 11052757 TI - Antioxidative activity in the pericarp and seed of Japanese pepper (Xanthoxylum piperitum DC). AB - The antioxidative activity of the dried pericarp and seed of Japanese pepper was studied. The ethyl acetate extract from the pericarp and the methanol extract from the seed showed strong antioxidative activity against linoleic acid by the ferric thiocyanate and thiobarbituric acid (TBA) methods. Japanese pepper contained 3.9 and 2.9 mg/100 g of dry weight (dw) of tocopherols in the pericarp and seed, respectively, alpha-Toc in the former constituting 82% of total tocopherol and gamma-Toc in the latter constituting 96%. Arbutin and magnoflorine were isolated as antioxidants and their chemical structures determined by instrumental analyses. The contents of arbutin evaluated as the trifluoroacetate derivative by GC-MS were 35 and 3.0 mg/100 g of dw in the pericarp and seed, respectively. Magnoflorine was present only in the seed, and not in the pericarp. Both arbutin and magnoflorine exhibited antioxidative activity against linoleic acid and radical-scavenging activity against the DPPH radical. PMID- 11052758 TI - beta-glucosidases from five black Aspergillus species: study of their physico chemical and biocatalytic properties. AB - Five black Aspergillus strains (A. aculeatus, A. foetidus, A. japonicus, A. niger, and A. tubingensis) were cultivated on crude wheat arabinoxylan as the carbon source under defined pH, temperature, and oxygen conditions. Protein and beta-glucosidase content differed remarkably within the obtained culture filtrates, of which eleven beta-glucosidases were isolated. Seven beta glucosidases were purified to apparent electrophoretic homogeneity using anion exchange and gel-permeation chromatography. They were found to be acidic proteins and most of them appeared to be glycoproteins with a molecular mass between 93 and 142 kDa. Classification of the beta-glucosidases into four groups (I-A, I-B, II, and III) is suggested according to their physicochemical and biocatalytic properties. The major beta-glucosidases were assigned to groups I-A and I-B, the minor beta-glucosidases to groups II and III, comprising acid-tolerant and glucose-tolerant enzymes, respectively. PMID- 11052759 TI - Solubility, tensile, and color properties of modified soy protein isolate films. AB - Protein solubility (PS) values of different soy protein isolate (SPI) films were determined in water, 0.01 N HCl, 0.01 N NaOH, 4 M urea, and 0.2 M 2 mercaptoethanol. Tensile and color (L, a, and b values) properties of films also were determined. Control films were cast from heated (70 degrees C for 20 min), alkaline (pH 10) aqueous solutions of SPI (5 g/100 mL of water) and glycerin (50% w/w of SPI). Additional films were cast after incorporation of dialdehyde starch (DAS) at 10% w/w of SPI or small amounts of formaldehyde in the film-forming solutions. Also, control film samples were subjected to heat curing (90 degrees C for 24 h), UV radiation (51.8 J/m(2)), or adsorption of formaldehyde vapors. PS of control films was highest (P < 0.05) in 2-mercaptoethanol, confirming the importance of disulfide bonds in SPI film formation. All treatments were effective in reducing (P < 0.05) film PS in all solvents. Both DAS and adsorbed formaldehyde rendered the protein in films practically insoluble in all solvents. Adsorption of formaldehyde vapors and heat curing also substantially increased (P < 0.05) film tensile strength from 8.2 to 15.8 or 14.7 MPa, respectively. However, heat curing decreased (P < 0.05) film elongation at break from 30 to 6%. Most treatments had small but significant (P < 0.05) effects on b color values, with DAS-containing films having the greatest (P < 0. 05) mean b value (most yellowish). Also, DAS-containing, heat-cured, and UV-irradiated films were darker, as evidenced by their lower (P < 0.05) L values, than control films. It was demonstrated that PS of SPI films can be notably modified through chemical or physical treatments prior to or after casting. PMID- 11052760 TI - Reaction rate modeling in cryoconcentrated solutions: alkaline phosphatase catalyzed DNPP hydrolysis. AB - The hydrolysis of disodium p-nitrophenyl phosphate catalyzed by alkaline phosphatase was chosen as a model to study the kinetics of changes in frozen food products. The initial reaction rate was determined in concentrated sucrose solutions down to -24 degrees C, and the enzymatic characteristics K(M) and V(max) were calculated. The experimental data were compared to the kinetics predicted by assuming that the reaction was viscosity dependent. Indeed, an analysis of the enzymatic reaction demonstrated that both the diffusion of the substrate and the flexibility of the enzyme segments were controlled by the high viscosity of the media. When the temperature was too low for the viscosity to be measured simply, the Williams-Landel-Ferry equation was used to predict the viscosity, taking, as reference temperature, the glass transition temperature (T(g)) corresponding to the concentration of the freeze-concentrated phase at the test temperature. Predicted values of the reaction rate were very close to the experimental ones in the studied temperature range. PMID- 11052761 TI - Molecular characterization around a glassy transition of starch using (1)H cross relaxation nuclear magnetic resonance. AB - The aim of this work was to characterize the glassy-rubbery transition in starch gels using molecular (NMR) techniques. Proton cross-relaxation ((1)H CR) NMR spectra of gelatinized starch ( approximately 50% mc) were obtained by cooling stepwise from 20 to -30 degrees C. A significant line broadening was observed in the CR spectra between 0 and -10 degrees C. Deconvolution of the spectra into its component curves (broad and narrow) yielded a peak amplitude, width at half height, and peak area for each curve. Between 0 and -10 degrees C (temperatures around T(g)), a significant line width change in the broad component (rigid solid) was apparent. These observed qualitative changes may be evidence of a glassy-rubbery transition at a molecular (short-range) level which are strengthened by a similar transition temperature range found previously with (13)C CP-MAS and DMA tan delta(T) measurements. However, the increase in the relative quantity of rigid protons observed by (1)H CR NMR spectra could also be attributed to ice. The (1)H CR NMR method showed its potential application for probing solid components in gels using a simple and economical NMR spectrometer, without the need for a solid-state instrument. PMID- 11052762 TI - Fluorescence of raw cane sugars evaluated by chemometrics. AB - In a fluorescence study of raw cane sugar samples, two-way and three-way chemometric methods have been used to extract information about the individual fluorophores in the sugar from fluorescence excitation-emission landscapes. A sample set of 47 raw sugar samples representing a varied selection was analyzed, and three individual fluorophores with (275, 350) nm, (340, 420) nm, and (390, 460) nm as their approximate excitation and emission maxima were found. The spectral profiles of the fluorophores were estimated with the three-way decomposition model PARAFAC. Two-way principal component analysis (PCA) of unfolded fluorescence landscapes confirmed the PARAFAC results and showed patterns of samples related to time of storage. Partial least squares (PLS) calibration models of color at 420 nm had a high model error due to the very high color range of the raw sugars, but variable selection performed on the fluorescence data revealed that all three fluorophores were correlated to color. The (275, 350) nm fluorophore is considered as a color precursor to the color developed on storage and the (340, 420) nm and (390, 460) nm fluorophores show colorant polymer characteristics. PMID- 11052763 TI - Oxidative in vitro metabolism of the soy phytoestrogens daidzein and genistein. AB - The oxidative metabolism of the major soy isoflavones daidzein and genistein was investigated using liver microsomes from Aroclor-treated male Wistar rats. Both daidzein and genistein were extensively metabolized and are therefore excellent substrates for cytochrome P450 enzymes. The identity of the metabolites was elucidated using high-performance liquid chromatography (HPLC) with diode array detection, gas chromatography-mass spectrometry (GC/MS), and HPLC/atmospheric pressure ionization electrospray mass spectrometry (API-ES MS) as well as reference substances. Daidzein was converted to nine metabolites, comprising four monohydroxylated, four dihydroxylated, and one trihydroxylated metabolite. Genistein was metabolized to four monohydroxylated and two dihydroxylated products. With both isoflavones the additional hydroxy groups are exclusively introduced into the ortho positions of existing phenolic hydroxy groups. The major metabolites of daidzein were identified as 6,7,4'-trihydroxyisoflavone, 6,7,3',4'-tetrahydroxyisoflavone, 7,8, 4'-trihydroxyisoflavone, and 5,6,7,4' tetrahydroxyisoflavone. The main microsomal metabolites of genistein were 5,6,7, 4'-tetrahydroxyisoflavone and 5,7,8,4'-tetrahydroxyisoflavone. Furthermore, the GC/MS and HPLC/API-ES MS analysis support the conclusion that one monohydroxylated metabolite of daidzein and genistein is hydroxylated at the aliphatic position C-2 of the C-ring. The UV-vis, GC/MS, and HPLC/MS data of all detected metabolites as well as the derived chemical structure of the metabolites are presented. Most metabolites are reported in this paper for the first time. On the basis of these findings it is suggested that hydroxylation reactions may also play an important role in the in vivo metabolism of the soy isoflavones daidzein and genistein. PMID- 11052764 TI - Lactones. 9. Synthesis of terpenoid lactones-active insect antifeedants. AB - Starting from (+)- and (-)-perillyl alcohols, via Claisen rearrangement and iodolactonization, four enantiomeric pairs of gamma-lactones were obtained. The structures of compounds were established by both spectroscopic and crystallographic methods. The lactones were tested for antifeeding activity toward grain storage pests-the granary weevil beetle (Sitophilus granarius L.), the khapra beetle (Trogoderma granarium Ev.), and the confused flour beetle (Tribolium confusum Duv.). The results of the tests proved that two compounds, (1R,4R, 6R)-(-)-4-(1-methylethenyl)-9-oxabicyclo[4.3.0]nonan-8-one (8a) and its enantiomer (8b), are very active antifeedants against all of the above tested species. The lactone 8b is also active against the peach-potato aphid (Myzus persicae Sulz.). The activity depends on the absolute configurations of compound chiral centers. Additionally, the lactones 8a and 8b are characterized by interesting fragrances. PMID- 11052765 TI - Glutathione reductase in wheat grain. 1. Isolation and characterization. AB - Durum wheat (Triticum durum, Desf.) endosperm of mature kernels contained a single form of glutathione reductase (GR); it appeared about the 18th day after anthesis while another isoform, present at the early stages of grain development, disappeared between the 20th and 30th days after flowering. The form that was present at grain maturity was isolated and characterized. It was composed of two monomers, each one having an apparent molecular mass of about 60 kDa. The K(m) values for NADPH and for GSSG were 3.7 and 9.1 microM, respectively, and the V(m) values for NADPH and for GSSG were 594 and 575 microkat.mg(-)(1) protein, respectively. The pH(i) of the enzyme was situated between pH 4.4 and 4.5. At a constant temperature of 25 degrees C, the optimum GR activity was found to be between pH 7.5 and 8.0. It was relatively resistant to high temperatures and was very resistant to very low temperatures. PMID- 11052766 TI - Lactoferrin in infant formulas: effect on oxidation. AB - Lactoferrin is an iron transport protein present in human milk at an average concentration of 1.4 mg/mL. Commercially modified infant formulas based on cow's milk contain much lower amounts of lactoferrin (0.1 mg/mL lactoferrin) and soy based formulas have none. In addition to its role in iron transport, lactoferrin has bacteriostatic and bactericidal activities. Infant formulas are supplemented with relatively large amounts of iron (up to 12 mg/L). The effect of various concentrations of added lactoferrin and supplemental iron on lipid oxidation was tested in two different infant formulas. The extent of oxidation in the formulas as a function of time was determined by formation of hydroperoxides, production of hexanal, and fluorescence. On the basis of all three of these determinations, lactoferrin acted as an antioxidant in the absence and presence of different concentrations of supplemented iron. Lactoferrin inhibited oxidation in a concentration-dependent manner even at concentrations beyond its capacity to bind iron at its two high affinity binding sites. Lactoferrin can be used, therefore, as a dual purpose additive in infant formulas and similar food products for its antioxidant and its antimicrobial properties. PMID- 11052767 TI - Purification and characterization of an esterase conferring resistance to fenitrothion in Oryzaephilus surinamensis (L.) (insecta, coleoptera, silvanidae). AB - Esterases from a fenitrothion-resistant strain (VOSF) of the saw-toothed grain beetle, Oryzaephilus surinamensis (L.), are presumed to play a role in conferring resistance to malathion, fenitrothion, and chlorpyrifos-methyl. Colorimetric assays showed a significant positive correlation between increased resistance to fenitrothion in strains of O. surinamensis examined and elevated esterase hydrolytic activity to substrates of p-nitrophenyl acetate, alpha-naphthyl acetate, and beta-naphthyl acetate. Esterase zymograms showed different banding patterns between VOSF and an insecticide-susceptible strain, VOS48. A major esterase in the VOSF strain, not detected in VOS48, was purified and characterized by chromatographic and electrophoretic techniques. On the basis of SDS-polyacrylamide gel eletrophoresis, the molecular mass of the purified esterase from VOSF was 130 kDa and consisted of two 65 kDa subunits. Additional properties of this enzyme are discussed. PMID- 11052768 TI - Honeys from different floral sources as inhibitors of enzymatic browning in fruit and vegetable homogenates. AB - Honeys from different floral sources were evaluated for their antioxidant content and for their ability to inhibit enzymatic browning in fruits and vegetables. Antioxidant contents of honeys vary widely from different floral sources, as do their abilities to protect against enzymatic browning. Polyphenol oxidase (PPO) activity was reduced over a range of approximately 2-45% in fruit and vegetable homogenates, corresponding to a reduction in browning index by 2.5-12 units. Soy honey was particularly effective when compared to clover honey, which had a similar antioxidant content. When compared to commercial inhibitors of browning, honeys were less effective; however, in combination they added to the effectiveness of metabisulfite and ascorbic acid. Honey has great potential to be used as a natural source of antioxidants to reduce the negative effects of PPO browning in fruit and vegetable processing. PMID- 11052769 TI - Formation of disulfide bonds in acid-induced gels of preheated whey protein isolate. AB - Cold gelation of whey proteins is a two-step process. First, protein aggregates are prepared by a heat treatment of a solution of native proteins in the absence of salt. Second, after cooling of the solution, gelation is induced by lowering the pH at ambient temperature. To demonstrate the additional formation of disulfide bonds during this second step, gelation of whey protein aggregates with and without a thiol-blocking treatment was studied. Modification of reactive thiols on the surface of the aggregates was carried out after the heat-treatment step. To exclude specific effects of the agent itself, different thiol-blocking agents were used. Dynamic light scattering and SDS-agarose gel electrophoresis were used to show that the size of the aggregates was not changed by this modification. The kinetics of gelation as determined by the development of pH and turbidity within the first 8 h of acidification were not affected by blocking thiol groups. During gelation, formation of large, covalently linked, aggregates occurred only in the case of unblocked WPI aggregates, which demonstrates that additional disulfide bonds were formed. Results of permeability and confocal scanning laser microscope measurements did not reveal any differences in the microstructure of networks prepared from treated or untreated whey protein aggregates. However, gel hardness was decreased 10-fold in gels prepared from blocked aggregates. Mixing different amounts of blocked and unblocked aggregates allowed gel hardness to be controlled. It is proposed that the initial microstructure of the gels is primarily determined by the acid-induced noncovalent interactions. The additional covalent disulfide bonds formed during gelation are involved in stabilizing the network and increase gel strength. PMID- 11052770 TI - Effects of ozone and oxygen on the degradation of carotenoids in an aqueous model system. AB - The effects of ozone and oxygen on the degradation of carotenoids in an aqueous model system were studied. All-trans beta-carotene, 9-cis beta-carotene, beta cryptoxanthin, and lycopene were adsorbed onto a C(18) solid phase and exposed to a continuous flow of water saturated with oxygen or ozone at 30 degrees C. Carotenoids were analyzed using HPLC with a C(30) column and a photodiode array detector. Approximately 90% of all-trans beta-carotene, 9-cis beta-carotene, and beta-cryptoxanthin were lost after exposure to ozone for 7 h. A similar loss of lycopene occurred in only 1 h. When exposed to oxygen, all carotenoids, except beta-cryptoxanthin, degraded at lower rates. The degradation of all the carotenoids followed zero-order reaction kinetics with the following relative rates: lycopene > beta-cryptoxanthin > all-trans beta-carotene > 9-cis beta carotene. The major degradation products of beta-carotene were tentatively identified on the basis of their elution on the HPLC column, UV-Vis spectra, and electrospray LC-MS. Predominant isomers of beta-carotene were 13-cis, 9-cis, and a di-cis isomer. Products resulting from cleavage of the molecule were beta-apo 13-carotenone and beta-apo-14'-carotenal, whereas epoxidation yielded beta carotene 5,8-epoxide and beta-carotene 5, 8-endoperoxide. PMID- 11052771 TI - Purification and biochemical properties of dipeptidyl peptidase I from porcine skeletal muscle. AB - Dipeptidyl peptidase I (DPP I; EC 3.4.14.1) was purified from porcine skeletal muscle after several steps such as heat treatment, ammonium sulfate fractionation, gel filtration chromatography, and HPLC anion exchange chromatography. The purified enzyme showed a native molecular mass of approximately 200 kDa on Sephacryl S-200 column chromatography. Two protein bands of 65 and 42 kDa were obtained by SDS-PAGE, indicating its oligomeric nature. Maximum activity was reached at pH 5.5 and 55 degrees C. DPP I shared some common substrate specificities, both on synthetic derivatives and on real peptides, with porcine muscle DPP III. The enzyme required reducing agents for full activation, although the halide requirement was not proved. DPP I was inhibited by the assayed cysteine peptidase inhibitors except p-CMB. The serine peptidase inhibitor 3, 4-DCI also inhibited the enzyme as did the divalent cations Co(2+), Mn(2+), and Zn(2+). On the basis of its properties, DPP I may contribute to the generation of dipeptides during the processing of meat and/or meat products, including cooked ham. PMID- 11052772 TI - Purification and structural characterization of LTP1 polypeptides from beer. AB - We report on the purification of lipid transfer proteins (LTP) from barley seeds and beer with the aim of investigating the chemical modifications that occur during the brewing process. In seeds, the well-known LTP of 9 kDa (LTP1) has been found together with a second form named LTPb that displays comparable amino acid composition but was not fully sequenced. These two forms have been recovered in beer with marked chemical modifications including disulfide bond reduction and rearrangement and especially glycation by Maillard reaction. The glycation is heterogeneous with variable amounts of hexose units bound to LTPs. Circular dichroism shows that glycated LTP1 having all their disulfide bridges reduced are totally unfolded. These results provide a first basis for understanding how barley LTPs become foam-promoting agents during the malting and brewing process. PMID- 11052773 TI - Comparison of antioxidant and antimicrobial activities of tilia (Tilia argentea Desf ex DC), sage (Salvia triloba l.), and black tea (Camellia sinensis) extracts. AB - The antioxidant activity of the water extract of Tilia argentea Desf ex DC was determined by the thiocyanate method. The antioxidant activity of the water extract increased with the increasing amount of lyophilized extract (50-400 microg) added into the linoleic acid emulsion. Statistically significant effect was determined in 100 microg and higher amounts. Antioxidant activities of water extracts of tilia (Tilia argentea Desf ex DC), sage (Salvia triloba L.), and two Turkish black teas commercially called Rize tea and young shoot tea (Camellia sinensis) were compared. For comparison studies, 100 microg portions of extracts were added into test samples. All samples were able to show statistically significant antioxidant effect. Both of the tea extracts showed highest antioxidant activities, nevertheless, differences between tilia and sage and tilia and tea were not statistically significant (for both cases p > 0.05). Like antioxidant activity, the reducing power of water extract of Tilia argentea Desf ex DC was also concentration dependent. Even in the presence of 50 microg of extract, the reducing power was significantly higher than that of the control (p < 0.05) in which there was no extract. Unlike antioxidant activity, the highest reducing power activity was shown by sage extract. Among the tea extracts, young shoot extract was the most effective one, however, it had significantly lower activity than sage (p < 0.05). Although tea flower had the lowest reducing power activity, it was higher than that of tilia. But this difference was not statistically significant (p > 0.05). From these results, we could suggest that although the reducing power of a substance may be an indicator of its potential antioxidant activity, there may not always be a linear correlation between these two activities. In addition, antimicrobial activities of each of the above extracts were studied by disk diffusion methods on different test microorganisms. None of the extracts showed antibacterial activity on the studied microorganisms. PMID- 11052774 TI - Toward the authentication of varietal wines by the analysis of grape (Vitis vinifera L.) residual DNA in must and wine using microsatellite markers. AB - In an attempt to develop a technique for the identification of grape cultivars in commercial wines, a method for the extraction of DNA from must and experimental wines was adapted and optimal PCR conditions for the amplification of this DNA were established. DNA was analyzed during the fermentation process for six cultivars (Chardonnay, Clairette blanche, Grenache noir, Merlot, Muscat blanc a petits grains, and Syrah). The extractions were performed on solid parts in suspension as well as on the aqueous fraction. Expected profiles for these cultivars were obtained with DNA extracted from the solid parts during all of the fermentation process and for the wine. The analysis of DNA extracted from aqueous fractions was less reproducible, and microsatellite amplifications were obtained only in the case of Clairette blanche, Merlot, and Syrah wines. Results demonstrate that the purification process is adequate for the analysis but that the DNA concentration represents the main limiting factor. Technical improvements of the method are discussed. PMID- 11052775 TI - Latent polyphenol oxidases from sago log (Metroxylon sagu): partial purification, activation, and some properties. AB - Latent polyphenol oxidase (LPPO), an enzyme responsible for the browning reaction of sago starches during processing and storage, was investigated. The enzyme was effectively extracted and partially purified from the pith using combinations of nonionic detergents. With Triton X-114 and a temperature-induced phase partitioning method, the enzyme showed a recovery of 70% and purification of 4. 1 fold. Native PAGE analysis of the partially purified LPPO revealed three activity bands when stained with catechol and two bands with pyrogallol. The molecular masses of the enzymes were estimated by SDS-PAGE to be 37, 45, and 53 kDa. The enzyme showed optimum pH values of 4.5 with 4-methylcatechol as a substrate and 7.5 with pyrogallol. The LPPO was highly reactive toward diphenols and triphenols. The activity of the enzyme was greatly enhanced in the presence of trypsin, SDS, ethanol, and linoleic acid. PMID- 11052776 TI - Effects of sugars on whey protein isolate gelation. AB - Whey protein isolate (WPI) gels were prepared from solutions containing ribose or lactose at pH values ranging from 6 to 9. The gels with added lactose had no color development, whereas the gels with added ribose were orange/brown. Lactose stabilized the WPI to denaturation, which increased the time and temperature required for gelation, thus decreasing the fracture modulus of the gel compared to the gels with added ribose and the gels with no sugar added. Ribose, however, favored the Maillard reaction and covalent cross-linking of proteins, which increased gel fracture modulus. The decreased pH caused by the Maillard reaction in the gels containing ribose occurred after protein denaturation and gelation, thus having little if any effect on the gelation process. PMID- 11052777 TI - Addition of sugars influences color of oil-in-water emulsions. AB - The influence of glucose, fructose, lactose, and glycerol on the color and appearance of surfactant-stabilized oil-in-water emulsions containing a red dye was investigated. A stabilized (Tween-20) oil-in-water emulsion was diluted into sugar solutions to give a range of oil droplet and sugar concentrations. Tristimulus coordinates (L, a, b) and reflectance spectra were measured using a spectrophotometer. With increasing sugar concentration, reflectance spectra shifted to lower reflectance values. Tristimulus coordinates were reduced by approximately 50% for emulsions containing high concentrations of sugar. Adding fructose to emulsions reduced L, a, b values more significantly than adding glucose, lactose, or glycerol. Tristimulus coordinates remained constant when the temperature was raised from 20 to 80 degrees C. The experimental results were explained in terms of the change of relative refractive index at the water-oil interface. The results have important implications for the food industry as they offer a new means to control and optimize the color of food emulsions. PMID- 11052778 TI - Decreased superoxide anion production in cultured human promonocyte cells (THP-1) due to polyphenol mixtures from olive oil processing wastewaters. AB - The purpose of this study was to examine whether human monocytic line THP-1 after differentiation into adherent macrophages, taken as a model of human macrophages implicated in atheroma, is able to produce lower quantities of O(2)(*)(-) either in the presence of polyphenol-rich olive oil wastewater (OWW) fractions or after OWW preincubation and withdrawal from the medium. In these respective conditions, the purpose was to examine the scavenging activity and the cell action of OWW toward O(2)(*)(-) production. It was clearly seen that OWW fractions lowered the O(2)(*)(-) production in both conditions, leading to the conclusion that they were able to scavenge O(2)(*)(-) and to depress O(2)(*)(-) production in the cell. Given the role of O(2)(*)(-) in LDL oxidation and oxidized LDL in atheroma, these results support an antiatherogenic role of OWW and its potential utilization as a food complement. PMID- 11052779 TI - Antioxidative activity of green tea treated with radical initiator 2, 2'-azobis(2 amidinopropane) dihydrochloride. AB - This study investigated the antioxidative activity of green tea extract, and a green tea tannin mixture and its components, under conditions of radical generation using the hydrophilic azo compound, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) to generate peroxyl radicals at a constant and measurable rate in the cultured renal epithelial cell line, LLC-PK(1), which is susceptible to oxidative damage. Treatment with AAPH decreased cell viability and increased the formation of thiobarbituric acid-reactive substances. However, green tea extract, and the tannin mixture and its components, comprising (-) epigallocatechin 3-O-gallate (EGCg), (-)-gallocatechin 3-O-gallate (GCg), (-) epicatechin 3-O-gallate (ECg), (-)-epigallocatechin (EGC), (+)-gallocatechin (GC), (-)-epicatechin (EC), and (+)-catechin (C), showed protective activity against AAPH-induced cellular damage. The tannin mixture and its components exhibited higher antioxidative activity than the green tea extract. Furthermore, EGCg and GCg had higher activity than EGC and GC, respectively. In particular, EGCg exerted the most significant cellular protective activity against AAPH. These results indicate that green tea tannin may inhibit cellular loss and lipid peroxidation resulting from the peroxyl radical generated by AAPH, and that the chemical structure of tannin is also involved in the activity, suggesting that the O-dihydroxy structure in the B ring and the galloyl groups are important determinants for radical scavenging and antioxidative potential. PMID- 11052780 TI - Antimutagenic activity of cacao: inhibitory effect of cacao liquor polyphenols on the mutagenic action of heterocyclic amines. AB - We investigated the effect of polyphenols derived from cacao liquor on the mutagenic action of heterocyclic amines (HCAs) in vitro and ex vivo. In the Ames test, the cacao liquor polyphenols showed antimutagenic effects in bacteria treated with HCA in the presence of an S-9 mixture; however, they showed less efficacy than quercetin. On the other hand, the cacao liquor polyphenols showed potent antimutagenic activity in bacteria treated with activated forms of HCA, compared with quercetin. We also evaluated the effect of these compounds on enzymatic activation of HCA. They weakly suppressed the production of activated HCA. In the host-mediated assay in mice, a method used to estimate the potential carcinogenicity of chemicals ex vivo, oral administration of the cacao liquor polyphenols, reduced the number of colonies of revertant bacteria recovered from the liver. These data suggest that the cacao liquor polyphenols have an antimutagenic effect not only in vitro, but also ex vivo. PMID- 11052781 TI - Production of polyclonal antibodies against ochratoxin A and its detection in chilies by ELISA. AB - Polyclonal antibodies were produced for Ochratoxin A (OA) by injecting OA-bovine serum albumin (BSA) conjugate subcutaneously at multiple sites into a New Zealand White inbred rabbit. Antiserum could be used at a dilution exceeding 1:100 000 in an indirect competitive enzyme-linked immunosorbent assay (ELISA), and detected OA concentrations up to 0.1 ng/mL. The 50% inhibition binding (I(50)) of OA was 5 ng/mL. Antibodies did not react with ochratoxin B, coumarin, 4-hydroxycoumarin, L phenylalanine, and aflatoxin B1. OA contamination in chilies (Capsicum annum L.) collected from commercial markets and cold storage units was determined. The mean recoveries from OA-free chilies spiked with 1 to100 microg of OA per kg of chili sample were 90-110% with a standard deviation of <10%. Of 100 chili samples tested, 26 were found to contain over 10 microg/kg of OA. In 12 samples the OA concentration varied from 10 to 30 microg/kg, in 10 samples from 30 to 50 microg/kg, in 3 samples from 50 to100 microg/kg, and in one sample it was 120 microg/kg. This is the first record in India of OA in chilies, a major component of cooked foods in this country, and it is noteworthy that OA contamination exceeded the permissible limit for human consumption of less than 20 microg/kg in over 26% of the market samples tested. PMID- 11052782 TI - Polycyclic aromatic hydrocarbons in liquid smoke flavorings obtained from different types of wood. Effect of storage in polyethylene flasks on their concentrations. AB - Smoke flavorings are widely used as an alternative to the traditional smoking techniques. Smoke generation conditions can determine the level of polycyclic aromatic hydrocarbons (PAHs) in the smoke and, consequently, in these preparations. In this paper, the influence of the wood source on the formation of PAHs is studied. For this purpose, five liquid smoke flavorings, obtained from different types of wood, were used. Sample aliquots, including deuterated internal standards, were subjected to an alkaline treatment, extracted by liquid liquid partition and cleaned up by means of silica tubes, followed by gas chromatography-mass spectrometry analysis. The results reveal that the flavoring obtained from poplar wood presents the highest number and concentrations of both total and carcinogenic PAHs, even though the levels of these latter are very low. It has also been observed that the storage of smoke flavorings in polyethylene flasks reduces the concentration of some PAHs. PMID- 11052783 TI - Investigation on the possible formation of N-nitroso-N-methylurea by nitrosation of creatinine in model systems and in cured meats at gastric pH. AB - N-Nitroso-N-methylurea (NMU) is a highly potent direct-acting carcinogen that has been shown to induce cancer in a number of animal species. Although previous research has indicated that nitrosation of creatinine (CRN), a common constituent of meats, dried fish, and seafoods, can form traces of NMU, there is uncertainty as to (1) the yield of NMU and (2) whether detectable amounts of NMU can be formed from cured meats following nitrosation under acidic conditions given the low residual levels of nitrite found in cured meats at the present time. Lack of sensitive and specific analytical methods most likely has hindered progress in research in these areas. An HPLC postcolumn denitrosation-thermal energy analyzer technique and a GC-MS confirmation technique were developed for the determination of NMU in cured meats. Both techniques are highly sensitive (0.5 and 0.03 ppb, respectively) and specific. The optimum pH for NMU formation from CRN ranged between pH 1 and pH 3, and the yields of NMU under variable reactant concentrations ranged between 0.00004 and 0.0046%. When 27 samples of various cured meats (10 g aliquots each) were acidified with HCl (final pH values of 0.8 2.5) and incubated at room temperature for 2 h, without any additional nitrite, 24 gave results below detectable levels but 3 formed 2-26 ng of NMU/10 g of meat. Incubation of the negative meats with additional nitrite (50-500 microg/g of meat) formed 0.6-176 ng of NMU/10 g of sample. Although the amounts of NMU formed were extremely small, this seems to be the first reported formation of NMU from cured meats with and without additional nitrite. PMID- 11052784 TI - Structure-based design of novel bicyclic nonpeptide inhibitors for the src SH2 domain. PMID- 11052785 TI - Hunting the emesis and efficacy targets of PDE4 inhibitors: identification of the photoaffinity probe 8-(3-azidophenyl)-6- [(4-iodo-1H-1 imidazolyl)methyl]quinoline (APIIMQ). PMID- 11052786 TI - 7-Chloro-4,5-dihydro-8-(1,2,4-triazol-4-yl)-4-oxo-1,2,4-triazolo[1, 5 a]quinoxaline-2- carboxylates as novel highly selective AMPA receptor antagonists. PMID- 11052787 TI - Synthesis of a substance P antagonist with a somatostatin scaffold: factors affecting agonism/antagonism at GPCRs and the role of pseudosymmetry. PMID- 11052788 TI - Discovery of a potent, orally bioavailable beta(3) adrenergic receptor agonist, (R)-N-[4-[2-[[2-hydroxy-2-(3-pyridinyl)ethyl]amino]ethyl]phenyl]-4-[4 -[4 (trifluoromethyl)phenyl]thiazol-2-yl]benzenesulfonamide. AB - As part of our investigation into the development of orally bioavailable beta(3) adrenergic receptor agonists, we have identified a series of pyridylethanolamine analogues possessing a substituted thiazole benzenesulfonamide pharmacophore that are potent human beta(3) agonists with excellent selectivity against other human beta receptor subtypes. Several of these compounds also exhibited an improved pharmacokinetic profile in dogs. For example, thiazole sulfonamide 2e (R = 4 F(3)C-C(6)H(4)) is a potent full beta(3) agonist (EC(50) = 3.6 nM, 94% activation) with >600-fold selectivity over the human beta(1) and beta(2) receptors, which also displays good oral bioavailability in several mammalian species, as well as an extended duration of action. PMID- 11052789 TI - Design, synthesis, and X-ray crystal structure of a potent dual inhibitor of thymidylate synthase and dihydrofolate reductase as an antitumor agent. AB - A novel N-?2-amino-4-methyl[(pyrrolo[2, 3-d]pyrimidin-5-yl)ethyl]benzoyl?-L glutamic acid (3a) was designed and synthesized as a potent dual inhibitor of thymidylate synthase (TS) and dihydrofolate reductase (DHFR) and as an antitumor agent. Compound 3b, the N7-benzylated analogue of 3a, was also synthesized as an antitumor agent. The synthesis of 3a was accomplished via a 12-step sequence which involved the synthesis of 2-amino-4-methylpyrrolo[2,3-d]pyrimidine (10) in 5 steps from 2-acetylbutyrolactone. Protection of the 2-amino group of 10 and regioselective iodination at the 5-position followed by palladium-catalyzed coupling afforded intermediate 14 which was converted to 3a by reduction and saponification. Similar synthetic methodology was used for 3b. X-ray crystal structure of the ternary complex of 3a, DHFR, and NADPH showed that the pyrrolo[2, 3-d]pyrimidine ring binds in a "2,4-diamino mode" in which the pyrrole nitrogen mimics the 4-amino moiety of 2,4-diaminopyrimidines. This is the first example of a classical pyrrolo[2,3-d]pyrimidine antifolate shown to have this alternate mode of binding to DHFR. Compounds 3a and 3b were more inhibitory than LY231514 against TS from Lactobacillus casei and Escherichia coli. Analogue 3a was also more inhibitory against DHFR from human, Toxoplasma gondii, and Pneumocystis carinii. Evaluation of 3a against methotrexate (MTX)-resistant cell lines with defined mechanisms indicated that cross-resistance of 3a was much lower than that of MTX. Metabolite protection studies and folylpoly-gamma glutamate synthetase studies suggest that the antitumor activity of 3a against the growth of tumor cells in culture is a result of dual inhibition of TS and DHFR. Compound 3a inhibited the growth of CCRF-CEM and FaDu cells in culture at ED(50) values of 12.5 and 7.0 nM, respectively, and was more active against FaDu cells than MTX. In contrast, compound 3b was inactive against both cell lines. Compound 3a was evaluated in the National Cancer Institute in vitro preclinical antitumor screening program and afforded IG(50) values in the nanomolar range against a number of tumor cell lines. PMID- 11052790 TI - Computational predictions of binding affinities to dihydrofolate reductase: synthesis and biological evaluation of methotrexate analogues. AB - The relative binding affinities to human dihydrofolate reductase of four new potential antifolates, containing ester linkages between the two aromatic systems, were estimated by free energy perturbation simulations. The ester analogue, predicted to exhibit the highest binding affinity to human dihydrofolate reductase, and a reference ester (more structurally related to methotrexate) were synthesized. As deduced from the measured IC(50) values, the calculated ranking of the ligands was correct although a greater difference in affinity was indicated by the experimental measurements. Among the new antifolates the most potent inhibitor exhibited a similar pharmacokinetic profile to methotrexate but lacked activity in a complex antiarthritic model in rat in vivo. PMID- 11052791 TI - Identification of novel inhibitors of urokinase via NMR-based screening. AB - Using an NMR-based screen, a novel class of urokinase inhibitors were identified that contain a 2-aminobenzimidazole moiety. The inhibitory potency of this family of inhibitors is similar to that of inhibitors containing a guanidine or amidine group. However, unlike previously described guanidino- or amidino-based inhibitors which have pK(a) values greater than 9.0, urokinase inhibitors containing a 2-aminobenzimidazole have pK(a) values of 7.5. Thus, 2 aminobenzimidazoles may have improved pharmacokinetic properties which could increase the bioavailability of inhibitors which contain this moiety. A crystal structure of one of the lead inhibitors, 2-amino-5-hydroxybenzimidazole, complexed with urokinase reveals the electrostatic and hydrophobic interactions that stabilize complex formation and suggests nearby subsites that may be accessed to increase the potency of this new series of urokinase inhibitors. PMID- 11052792 TI - Prediction of drug absorption using multivariate statistics. AB - Literature data on compounds both well- and poorly-absorbed in humans were used to build a statistical pattern recognition model of passive intestinal absorption. Robust outlier detection was utilized to analyze the well-absorbed compounds, some of which were intermingled with the poorly-absorbed compounds in the model space. Outliers were identified as being actively transported. The descriptors chosen for inclusion in the model were PSA and AlogP98, based on consideration of the physical processes involved in membrane permeability and the interrelationships and redundancies between available descriptors. These descriptors are quite straightforward for a medicinal chemist to interpret, enhancing the utility of the model. Molecular weight, while often used in passive absorption models, was shown to be superfluous, as it is already a component of both PSA and AlogP98. Extensive validation of the model on hundreds of known orally delivered drugs, "drug-like" molecules, and Pharmacopeia, Inc. compounds, which had been assayed for Caco-2 cell permeability, demonstrated a good rate of successful predictions (74-92%, depending on the dataset and exact criterion used). PMID- 11052793 TI - New diarylmethylpiperazines as potent and selective nonpeptidic delta opioid receptor agonists with increased In vitro metabolic stability. AB - Nonpeptide delta opioid agonists are analgesics with a potentially improved side effect and abuse liability profile, compared to classical opioids. Andrews analysis of the NIH nonpeptide lead SNC-80 suggested the removal of substituents not predicted to contribute to binding. This approach led to a simplified lead, N, N-diethyl-4-[phenyl(1-piperazinyl)methyl]benzamide (1), which retained potent binding affinity and selectivity to the human delta receptor (IC(50) = 11 nM, mu/delta = 740, kappa/delta > 900) and potency as a full agonist (EC(50) = 36 nM) but had a markedly reduced molecular weight, only one chiral center, and increased in vitro metabolic stability. From this lead, the key pharmacophore groups for delta receptor affinity and activation were more clearly defined by SAR and mutagenesis studies. Further structural modifications on the basis of 1 confirmed the importance of the N, N-diethylbenzamide group and the piperazine lower basic nitrogen for delta binding, in agreement with mutagenesis data. A number of piperazine N-alkyl substituents were tolerated. In contrast, modifications of the phenyl group led to the discovery of a series of diarylmethylpiperazines exemplified by N, N-diethyl-4-[1-piperazinyl(8 quinolinyl)methyl]benzamide (56) which had an improved in vitro binding profile (IC(50) = 0.5 nM, mu/delta = 1239, EC(50) = 3.6 nM) and increased in vitro metabolic stability compared to SNC-80. PMID- 11052794 TI - N,N-Diethyl-4-(phenylpiperidin-4-ylidenemethyl)benzamide: a novel, exceptionally selective, potent delta opioid receptor agonist with oral bioavailability and its analogues. AB - The design, synthesis, and pharmacological evaluation of a novel class of delta opioid receptor agonists, N, N-diethyl-4-(phenylpiperidin-4 ylidenemethyl)benzamide (6a) and its analogues, are described. These compounds, formally derived from SNC-80 (2) by replacing the piperazine ring with a piperidine ring containing an exocyclic carbon carbon double bond, were found to bind with high affinity and exhibit excellent selectivity for the delta opioid receptor as full agonists. 6a, the simplest structure in the class, exhibited an IC(50) = 0.87 nM for the delta opioid receptors and extremely high selectivity over the mu receptors (mu/delta = 4370) and the kappa receptors (kappa/delta = 8590). Rat liver microsome studies on a selected number of compounds show these olefinic piperidine compounds (6) to be considerably more stable than SNC-80. This novel series of compounds appear to interact with delta opioid receptors in a similar way to SNC-80 since they demonstrate similar SAR. Two general approaches have been established for the synthesis of these compounds, based on dehydration of benzhydryl alcohols (7) and Suzuki coupling reactions of vinyl bromide (8), and are herewith reported. PMID- 11052795 TI - Synthesis and antiviral activity of oxaselenolane nucleosides. AB - As dioxolane and oxathiolane nucleosides have exhibited promising antiviral and anticancer activities, it was of interest to synthesize isoelectronically substituted oxaselenolane nucleosides, in which the 3'-CH(2) is replaced by a selenium atom. To study structure-activity relationships, various pyrimidine and purine oxaselenolane nucleosides were synthesized from the key intermediate, (+/ )-2-benzoyloxymethyl-1,2-oxaselenolane 5-acetate (6). Among the synthesized racemic nucleosides, cytosine and 5-fluorocytosine analogues exhibited potent anti-HIV and anti-HBV activities. It was of interest to obtain the enantiomerically pure isomers to determine if they have differential antiviral activities. However, due to the difficult and time-consuming nature of enantiomeric synthesis, a chiral HPLC separation was performed to obtain optical isomers from the corresponding racemic mixtures. Each pair of enantiomers of Se ddC and Se-FddC was separated by an amylose chiral column using a mobile phase of 100% 2-propanol. The results indicate that most of the anti-HIV activity of both cytosine and fluorocytosine nucleosides resides with the (-)-isomers. PMID- 11052797 TI - trans-1-[(2-Phenylcyclopropyl)methyl]-4-arylpiperazines: mixed dopamine D(2)/D(4) receptor antagonists as potential antipsychotic agents. AB - The dopaminergic receptor profile of a series of trans-1-[(2 phenylcyclopropyl)methyl]-4-arylpiperazines was examined. Aromatic substitution patterns were varied with the goal of identifying a compound having affinities for the D(2) and D(4) receptors in a ratio similar to that observed for the atypical neuroleptic clozapine. The compounds (1S, 2S)-trans-1-[(2 phenylcyclopropyl)methyl]-4-(2, 4-dichlorophenyl)piperazine (5m) and (1S, 2S) trans-1-[(2-phenylcyclopropyl)methyl]-4-(2, 4-dimethylphenyl)piperazine (5t) were selected for functional antagonists at D(2) and D(4) receptors and had a D(2)/D(4) ratio approximating that of clozapine; they proved inactive in behavioral tests of antipsychotic activity. PMID- 11052796 TI - Radioiodinated N-(2-diethylaminoethyl)benzamide derivatives with high melanoma uptake: structure-affinity relationships, metabolic fate, and intracellular localization. AB - Several radioiodinated N-(dialkylaminoalkyl)benzamides have been used for planar scintigraphy and single-photon emission computed tomography (SPECT) of melanoma metastases. In a quest for improved melanoma uptake and tissue selectivity, structure-activity studies for N-(2-diethylaminoethyl)benzamides with variation of phenyl substituents were performed using C57Bl/6 mice bearing B16 melanoma. Compounds 2 (4-amino-5-bromo-N-(2-diethylaminoethyl)-3-[(131)I]iodo-2-methoxybenz amide) and 6 (4-acetamido-N-(2-diethylaminoethyl)-5-[(131)I]iodo-2 methoxybenzamid e) showed at 6 h post iv injection, for example, melanoma uptake of 16.6 and 23.2% ID/g, respectively (mean values, n = 3). Uptake was 3-5 times higher (P < 0.01) than observed with benzamides known from the literature and was probably facilitated by the relatively slow urinary excretion of 2 or 6. In contrast, analogues lacking either the MeO, Ac, AcNH, or Br substituents exhibited reduced tumor uptake and high urinary excretion of radioactivity in various benzamide metabolites. Uptake of radioiodinated benzamides in B16 melanoma is not mediated by a specific mechanism such as sigma-receptor binding. 2 and 6 exhibited similar melanoma uptake values but quite different sigma(1) receptor affinities of K(i) = 0.278 +/- 0.018 and 5.19 +/- 0.40 microM, respectively. Uptake studies with IMBA (N-(2-diethylaminoethyl)-3-[(131)I]iodo-4 methoxybenzamide) or BZA (N-(2-diethylaminoethyl)-4-[(131)I]iodobenzamide) showed that with increasing dose of unlabeled compound the measured uptake of label was unchanged (IMBA) or even enhanced (BZA) while receptor binding of label decreased. Differential and equilibrium density-gradient centrifugation revealed that most of the radioactivity from labeled IMBA was associated with fractions containing melanin granules. Thus, structure-activity studies indicate that blood clearance rates and metabolic stability are the main determinants for benzamide uptake in melanoma. The high uptake and slow clearance of 6 offer considerable potential for melanoma imaging in patients, and this compound may also prove to be useful for radionuclide therapy. PMID- 11052798 TI - Sequential cytotoxicity: a theory evaluated using novel 2-[4-(3-aryl-2 propenoyloxy)phenylmethylene]cyclohexanones and related compounds. AB - Five series of novel compounds were synthesized in order to evaluate the theory of sequential cytotoxicity which seeks to exploit the view that various cancer cells are particularly susceptible to successive attacks by cytotoxic agents. The compounds prepared were various 2-[4-(3-aryl-2 propenoyloxy)phenylmethylene]cyclohexanone s 1 and the related Mannich bases 2. In addition the analogues 3-5 lacking an olefinic bond in the ester group were also synthesized, which were predicted to be less cytotoxic than the compounds of series 1 and 2. The atomic charges at the potential sites for interaction with cellular constituents were determined by molecular modeling calculations. The biodata obtained from murine and human neoplastic cells revealed that the predictions made regarding the viability of the theory were fulfilled in approximately two-thirds of the cases indicating that further investigation of this hypothesis is warranted. In addition, the significant potencies of some of the Mannich bases toward human tumor cell lines, in particular coupled to their selective toxicity toward human leukemic and colon cancer cells, confirms their usefulness in serving as lead molecules for further development. A preliminary investigation into the mode of action of representative compounds revealed their ability to induce apoptosis and inhibit the biosyntheses of ribonucleic acid and proteins. PMID- 11052799 TI - Synthesis and evaluation of N,N-dialkyl enkephalin-based affinity labels for delta opioid receptors. AB - To develop affinity labels for delta opioid receptors based on peptide antagonists, the Phe(4) residues of N,N-dibenzylleucine enkephalin and N,N diallyl[Aib(2),Aib(3)]leucine enkephalin (ICI-174, 864) were substituted with either Phe(p-NCS) or Phe(p-NHCOCH(2)Br). A general synthetic method was developed for the conversion of small peptide substrates into potential affinity labels. The target peptides were synthesized using Phe(p-NH(2)) and a Boc/Fmoc orthogonal protection strategy which allowed for late functional group conversion of a p amine group in the peptides to the desired affinity labeling moieties. A key step in the synthesis was the selective deprotection of a Boc group in the presence of a tert-butyl ester using trimethylsilyl trifluoromethanesulfonate (TMS-OTf). The target peptides were evaluated in radioligand binding experiments in Chinese hamster ovary (CHO) cells expressing delta or mu opioid receptors. The delta receptor affinities of the N, N-dibenzylleucine enkephalin analogues were 2.5-10 fold higher than those for the corresponding ICI-174,864 analogues. In general, substitution at the para position of Phe(4) decreased binding affinity at both delta and mu receptors in standard radioligand binding assays; the one exception was N, N-dibenzyl[Phe(p-NCS)(4)]leucine enkephalin (2) which exhibited a 2-fold increase in affinity for delta receptors (IC(50) = 34.9 nM) compared to N,N dibenzylleucine enkephalin (IC(50) = 78.2 nM). The decreases in mu receptor affinities were greater than in delta receptor affinities so that all of the analogues tested exhibited significantly greater delta receptor selectivity than the unsubstituted parent peptides. Of the target peptides tested, only N, N dibenzyl[Phe(p-NCS)(4)]leucine enkephalin (2) exhibited wash-resistant inhibition of radioligand binding to delta receptors. To our knowledge, 2 represents the first peptide-based affinity label to utilize an isothiocyanate group as the electrophilic affinity labeling moiety. As a result of this study, enkephalin analogue 2 emerges as a potential affinity label useful for the further study of delta opioid receptors. PMID- 11052800 TI - Synthesis and antiviral activity of 4-benzyl pyridinone derivatives as potent and selective non-nucleoside human immunodeficiency virus type 1 reverse transcriptase inhibitors. AB - Several 4-benzyl analogues of 5-ethyl-6-methyl-4-(phenylthio)pyridin-2(1H)-ones were synthesized and evaluated for their anti-HIV-l activities. Key transformations include metalation at the 4-C-position of 5-ethyl-2-methoxy-6 methyl-3-pivaloylaminopyridine (5) and its coupling with benzyl bromide or benzaldehyde derivatives. Biological studies revealed that some of the new 4 benzylpyridinones show potent HIV-1 specific reverse transcriptase inhibitory properties. Compounds 14, 19, and 27, which inhibit the replication of HIV-1 in CEM-SS cells, with IC(50) values ranging from 0.2 to 6 nM are the most active compounds in this series. Biochemical studies showed that compound 27 strongly inhibited the activity of a recombinant HIV-1 RT. Moreover, the infectivity of isolated HIV-1 particles was severely decreased after exposure to compound 27. Although cross resistance is frequently observed between non-nucleoside reverse transcriptase inhibitors, compound 27 was capable of inhibiting a virus resistant to nevirapine with an IC(50) of 40 nM. PMID- 11052801 TI - Novel 7-substituted camptothecins with potent antitumor activity. AB - The natural alkaloid camptothecin is the lead compound of a new class of antitumor agents with a unique mechanism of action (i.e. inhibition of DNA topoisomerase I). The pharmacological interest of these agents has generated a large number of derivatives and analogues endowed with potent cytotoxic activity, two of them being in clinical use as antitumor drugs. We have synthesized a new series of camptothecins substituted in position 7 with an alkyl or alkenyl chain bearing cyano and/or carbethoxy groups. These compounds showed potent cytotoxic activity in vitro against the human non-small-cell lung carcinoma H460 cell line, most of them exhibiting IC(50) values in the 0.05-1 microM range, more active than topotecan used as a reference compound. In particular 7-cyano-20S camptothecin (5a) showed high in vitro cytotoxicity against a topotecan-resistant H460 cell subline (H460/TPT) and a cisplatin-resistant ovarian carcinoma subline (IGROV-1/Pt 1). In an in vivo evaluation of the antitumor activity, 5a appeared significantly more effective than topotecan in the H460 tumor model and comparable with topotecan in a small-cell lung carcinoma model and a colon carcinoma model. The efficacy and good tolerability of this compound increase interest for further preclinical development. PMID- 11052802 TI - The novel silatecan 7-tert-butyldimethylsilyl-10-hydroxycamptothecin displays high lipophilicity, improved human blood stability, and potent anticancer activity. AB - We describe the rational design and synthesis of B- and A, B-ring-modified camptothecins. The key alpha-hydroxy-delta-lactone pharmacophore in 7-tert butyldimethylsilyl-10-hydroxycamptothecin (DB-67, 14) displays superior stability in human blood when compared with clinically relevant camptothecin analogues. In human blood 14 displayed a t(1/2) of 130 min and a percent lactone at equilibrium value of 30%. The tert-butyldimethylsilyl group renders the new agent 25-times more lipophilic than camptothecin, and 14 is readily incorporated, as its active lactone form, into cellular and liposomal bilayers. In addition, the dual 7 alkylsilyl and 10-hydroxy substitution in 14 enhances drug stability in the presence of human serum albumin. Thus, the net lipophilicity and the altered human serum albumin interactions together function to promote the enhanced blood stability. In vitro cytotoxicity assays using multiple different cell lines derived from eight distinct tumor types indicate that 14 is of comparable potency to camptothecin and 10-hydroxycamptothecin, as well as the FDA-approved camptothecin analogues topotecan and CPT-11. In addition, cell-free cleavage assays reveal that 14 is highly active and forms more stable top1 cleavage complexes than camptothecin or SN-38. The impressive blood stability and cytotoxicity profiles for 14 strongly suggest that it is an excellent candidate for additional in vivo pharmacological and efficacy studies. PMID- 11052803 TI - Role of hydrophobic interactions in binding S-(N-aryl/alkyl-N hydroxycarbamoyl)glutathiones to the active site of the antitumor target enzyme glyoxalase I. AB - Hydrophobic interactions play an important role in binding S-(N-aryl/alkyl-N hydroxycarbamoyl)glutathiones to the active sites of human, yeast, and Pseudomonas putida glyoxalase I, as the log K(i) values for these mechanism-based competitive inhibitors decrease linearly with increasing values of the hydrophobicity constants (pi) of the N-aryl/alkyl substituents. Hydrophobic interactions also help to optimize polar interactions between the enzyme and the glutathione derivatives, given that the K(i) value for S-(N hydroxycarbamoyl)glutathione (pi = 0) with the human enzyme is 35-fold larger than the interpolated value for this compound obtained from the log K(i) versus pi plot. Computational studies, in combination with published X-ray crystallographic measurements, indicate that human glyoxalase I binds the syn conformer of S-(N-aryl-N-hydroxycarbamoyl)glutathiones in which the N-aryl substituents are in their lowest-energy conformations. These studies provide both an experimental and a conceptual framework for developing better inhibitors of this antitumor target enzyme. PMID- 11052804 TI - Novel histamine H(3)-receptor antagonists with carbonyl-substituted 4-(3 (phenoxy)propyl)-1H-imidazole structures like ciproxifan and related compounds. AB - Novel histamine H(3)-receptor antagonists possessing a 4-(3-(phenoxy)propyl)-1H imidazole structure generally substituted in the para-position of the phenyl ring have been synthesized according to Mitsunobu or S(N)Ar reactions. With in vitro and in vivo screening for H(3)-receptor antagonist potency, the carbonyl substituted derivatives proved to be highly active compounds. A number of compounds showed in vitro affinities in the subnanomolar concentration range, and the 4-hexanoyl (10) and 4-acetyl-3-methyl (29) substituted derivatives showed in vivo antagonist potencies of about 0.1 mg/kg after po administration. Many proxifans were also tested for their affinities at other histamine receptor subtypes thereby demonstrating their pronounced H(3)-receptor subtype selectivity. Since the cyclopropyl ketone derivative 14 (ciproxifan) had high affinity in vitro as well as high potency in vivo, it was selected for further studies in monkeys. It showed good oral absorption and long-lasting, dose dependent plasma levels making it a promising compound for drug development. PMID- 11052805 TI - Inhibitors of NF-kappaB and AP-1 gene expression: SAR studies on the pyrimidine portion of 2-chloro-4-trifluoromethylpyrimidine-5-[N-(3', 5' bis(trifluoromethyl)phenyl)carboxamide]. AB - We investigated the structure-activity relationship studies of N-[3, 5 bis(trifluoromethyl)phenyl][2-chloro-4-(trifluoromethyl)pyrimidin-5 yl]carboxamide (1), an inhibitor of transcription mediated by both NF-kappaB and AP-1 transcription factors, with the goal of improving its potential oral bioavailability. Compounds were examined for cell-based activity, were fit to Lipinski's rule of 5, and were examined for potential gastrointestinal permeability using the intestinal epithelial cell line, Caco-2. Selected groups were substituted at the 2-, 4-, and 5-positions of the pyrimidine ring using solution-phase combinatorial methodology. The introduction of a fluorine in the place of 2-chlorine of 1 resulted in a compound with comparable activity. However, other substitutions at the 2-position resulted in a loss of activity. The trifluoromethyl group at the 4-position could be replaced with a methyl, ethyl, chlorine, or phenyl without a substantial loss of activity. The carboxamide group at the 5-position is critical for activity. If it was moved to the 6-position, the activity was lost. The 2-methyl analogue of 1 (81) showed comparable in vitro activity and improved Caco-2 permeability compared to 1. PMID- 11052806 TI - Nitrosothiol esters of diclofenac: synthesis and pharmacological characterization as gastrointestinal-sparing prodrugs. AB - Despite its widespread use, diclofenac has gastrointestinal liabilities common to nonsteroidal antiinflammatory drugs (NSAIDs) that might be reduced by concomitant administration of a gastrointestinal cytoprotectant such as nitric oxide (NO). A series of novel diclofenac esters containing a nitrosothiol (-S-NO) moiety as a NO donor functionality has been synthesized and evaluated in vivo for bioavailability, pharmacological activity, and gastric irritation. All S-NO diclofenac derivatives acted as orally bioavailable prodrugs, producing significant levels of diclofenac in plasma within 15 min after oral administration to mice. At equimolar oral doses, S-NO-diclofenac derivatives (20a 21b) displayed rat antiinflammatory and analgesic activities comparable to those of diclofenac in the carrageenan-induced paw edema test and the mouse phenylbenzoquinone-induced writhing test, respectively. All tested S-NO diclofenac derivatives (20a-21b) were gastric-sparing in that they elicited markedly fewer stomach lesions as compared to the stomach lesions caused by a high equimolar dose of diclofenac in the rat. Nitrosothiol esters of diclofenac comprise a novel class of NO-donating compounds having therapeutic potential as nonsteroidal antiinflammatory agents with an enhanced gastric safety profile. PMID- 11052807 TI - [GdPCP2A(H(2)O)(2)](-): a paramagnetic contrast agent designed for improved applications in magnetic resonance imaging. AB - A novel ligand based on a pyridine-containing macrocycle bearing two acetic and one methylenephosphonic arms (PCP2A) has been synthesized. An efficient synthesis of PCP2A is based on the macrocyclization reaction between 2,6 bis(chloromethyl)pyridine and a 1,4, 7-triazaheptane derivative bearing a methylenephosphonate group on N-4. The Gd(III) complex of PCP2A displays characteristic properties which make it a very promising contrast agent for improved applications in magnetic resonance imaging. In fact it shows (i) a very high stability constant (log K(GdPCP2A) = 23.4) which should guarantee against the in vivo release of toxic free Gd(III) ions and free ligand molecules and (ii) a relaxivity that is about 2 times higher than the values reported for contrast agents currently used in the clinical practice. Its high relaxivity is the result of the presence of two water molecules in the inner coordination sphere and a significant contribution from water molecule(s) hydrogen bonded to the phosphonate group. Moreover, the inner sphere water molecules are involved in an exchange with the bulk water which is relatively fast. This property is important for the attainment of an even higher relaxivity once the molecular reorientation rate of the [GdPCP2A(H(2)O)(2)](-) moiety is lengthened by means of conjugation to a macromolecular substrate. PMID- 11052808 TI - Discovery of novel p-arylthio cinnamides as antagonists of leukocyte function associated antigen-1/intracellular adhesion molecule-1 interaction. 1. Identification of an additional binding pocket based on an anilino diaryl sulfide lead. AB - The interaction between leukocyte function-associated antigen-1 (LFA-1), a member of the beta(2)-integrin family of adhesion molecules, and intracellular adhesion molecule ICAM-1 (cd54) is thought to play a critical role in the inflammatory process. On the basis of an anilino diaryl sulfide screening lead 1, in combination with pharmacophore analysis of other screening hits, we have identified an adjacent binding pocket. Subsequently, a p-ethenylcarbonyl linker was discovered to be optimal for accessing this binding site. Solution-phase parallel synthesis enabled rapid optimization of the cinnamides for this pocket. In conjunction with fine-tuning of the diaryl substituents, we discovered a novel series of potent, nonpeptide inhibitors of LFA-1/ICAM-1 interaction, exemplified by A-286982 (28h), which has IC(50) values of 44 and 35 nM in an LFA-1/ICAM-1 binding assay and LFA-1-mediated cellular adhesion assay, respectively. PMID- 11052809 TI - Transcriptional regulation of the human interleukin 1beta gene by fibronectin: role of protein kinase C and activator protein 1 (AP-1). AB - Interleukin 1beta (IL-1beta) is a multifunctional polypeptide considered a key cytokine during inflammation. Fibronectin (FN), a matrix glycoprotein highly expressed in injured tissues, can induce expression of IL-1beta in human blood monocytic cells. Herein, we explore the intracellular signals and transcriptional mechanisms responsible for IL-1beta induction by FN using human promonocytic U937 cells transfected with the human IL-1beta promoter connected to a reporter gene. Exposure of transfected U937s to FN resulted in increased expression of the full length IL-1beta promoter. This effect, mediated via the alpha5beta1 integrin, was associated with activation of mitogen-activated protein kinases (MAPKs) and was abolished by pre-treatment of cells with Calphostin C, a specific inhibitor of protein kinase C (PKC) activation. Deletion analysis and co-transfection studies using consensus activator protein 1 (AP-1) oligonucleotides suggested that an AP 1 site present in the 5' end of the IL-1beta promoter was involved in the FN induced response. Finally, electrophoretic mobility shift assays showed that FN induced binding of AP-1, but not NF-kappaB. Together, these experiments demonstrate that FN binding to the alpha5beta1 integrin activates MAPK-dependent signal pathways, and results in the transcription of the IL-1beta promoter in U937 cells by activating PKC and inducing AP-1. PMID- 11052810 TI - Expression analysis and characterization of alternatively spliced transcripts of human IL-7Ralpha chain encoding two truncated receptor proteins in relapsed childhood all. AB - In the family of cytokines and cytokine receptors, alternative splicing of pre mRNA is a frequently observed process that generates different protein isoforms from a single genetic locus. The splicing-derived cytokine receptor protein isoforms are mostly soluble receptors or show alterations in their cytoplasmic domain. It is possible that receptor abnormalities or a pathological ratio of different isoforms may contribute to leukaemia by circumventing normal growth factor control or altering the balance of proliferation and differentiation. IL-7 plays a critical role in early stages of both B and T cell maturation. Moreover, it stimulates the expansion of mature T cells including anti-tumour reactive cells as well as a number of T and B cell malignancies underlining its potential importance for deregulated lymphoid proliferation and leukaemogenesis. Here, we present detailed data on the expression of the interleukin 7 receptor alpha chain (IL-7Ralpha) in leukaemic cells from 210 children with acute lymphoblastic leukaemia (ALL) and describe two novel alternatively spliced transcripts of human IL-7Ralpha coding for truncated receptor proteins which are still capable of binding IL-7. IL-7Ralpha mRNA expression was more frequent in more mature pre-B ALL [91% (30/33)] than in common [81% (81/100)] or pro-B ALL [64% (18/28)], or even in T ALL [64% (29/45)]. These results are in concordance with flow cytometric analyses on the proportion of IL-7Ralpha bearing cells among total blast cell population. Our results lead us to assume that splicing derived IL 7Ralpha isoforms play a potential role in modulating IL-7 signal transduction and might be important for the pathogenesis of leukaemia. PMID- 11052812 TI - A neutralizing monoclonal antibody specific for the dimer interface region of IL 8. AB - We have generated two mAbs, 6G4.2.5 and A5.12.14, that are similarly capable of neutralizing the biologic activity of wild-type IL-8. To characterize these antibodies further, their reactivity against a series of engineered IL-8 monomer and dimer variants was examined using a neutrophil degranulation assay. While 6G4.2.5 was found to block effectively the biologic activity of all variants regardless of their dimerization status, the results for A5.12.14 differed dramatically. A5.12.14 fully inhibited the agonist activity of one of the monomer variants, partially blocked the activity of another, and had no effect on the activity of two other variants. These results suggested that the binding epitope of A5.12.14 was being affected by the particular amino acid substitutions introduced into the dimer interface region of the variants to disfavor dimerization. If A5.12.14 indeed binds to the dimer interface region of IL-8, it could be predicted that this mAb would be unable to inhibit the activity of dimeric IL-8. This was confirmed in studies which showed that A5.12.14 had no demonstrable effect on the activity of a constitutively dimeric IL-8 variant. These studies represent the first example of a mAb specific for the dimerization status of IL-8. PMID- 11052811 TI - Regulation of IL-1-induced gingival collagenase gene expression by activator protein-1 (c-Fos/c-Jun). AB - Matrix metalloproteinase-1 is probably involved in the progression of periodontal disease. The aim of this study was to investigate whether IL-1beta stimulates the expression of the activator protein 1 (AP-1) transcription factor and, consequently, if the AP-1 transcription factor participates in the regulation of collagenase gene expression in human gingival fibroblast cells. In this study, we demonstrate that the concentration of the protein components of AP-1 transcription factor, c-Fos and c-Jun, is enhanced by IL-1beta both at mRNA and protein levels, utilizing Northern blot analysis, electrophoretic mobility gel shift assay and Western blot analysis. The IL-1beta stimulated the collagenase CAT and AP-1-CAT activities in a dose dependent manner with respect to the amount of DNA used in transfections. Further, overexpression of c-Fos and c-Jun proteins revealed a dose-dependent transcriptional activation of the collagenase promoter. These findings, coupled with the existence of AP-1 consensus DNA binding sites on the collagenase gene promoter, show that regulation of collagenase gene expression by IL-1beta involves the transcription factor AP-1 in gingival fibroblasts. PMID- 11052813 TI - Stimulatory effects of cartilage-derived morphogenetic proteins 1 and 2 on osteogenic differentiation of bone marrow stromal cells. AB - Cartilage-derived morphogenetic proteins 1 and 2 (CDMP-1 and CDMP-2) are members of the bone morphogenetic protein (BMP) family which play an important role in embryonic skeletal development. Throughout adult life, bone marrow-derived precursor cells maintain their ability to differentiate into osteoblasts in response to local growth factors. This study examines the osteogenic potential of CDMP-1, CDMP-2, BMP-6 and osteogenic protein 1 (OP-1) in bone marrow stromal cells (BMSC) and investigates the endogenous expression of CDMPs/BMPs and their respective activin receptor-like kinase (ALK) receptors. A 4-day exposure of BMSC to CDMP-1, CDMP-2, BMP-6, and OP-1 under serum-free conditions stimulated the progression of the osteogenic lineage in a dose-dependent manner as evaluated by alkaline phosphatase activity and osteocalcin synthesis. In contrast to the BMPs, CDMP-1 and especially CDMP-2 were significantly less osteogenic, as confirmed by Northern blot analysis. Moreover, BMSC were shown to express endogenously CDMP-2, BMP-2 to -6 and ALK-1, -2, -3, -5 and -6. Phenotypic characterization of BMSC by RT-PCR showed transcripts of the fat marker adipsin and the prechondrocytic marker procollagen type IIA; however, we were unable to detect the mature cartilage markers, procollagen type IIB and aggrecan, even after growth factor treatment. Our data indicate that CDMP-1, CDMP-2, BMP-6 and OP-1 enhance the osteogenic phenotype in BMSC, with CDMPs being clearly less osteogenic than BMPs. The endogenous expression of a variety of CDMPs/BMPs and their respective ALK receptors, suggests a possible involvement of these growth factors in the osteogenic differentiation of bone marrow progenitor cells. PMID- 11052814 TI - Role of type I interferons during macrophage activation by lipopolysaccharide. AB - Activation of macrophages by bacterial lipopolysaccharide (LPS) is accompanied by the secretion of type I interferons (IFNs) which can act in an autocrine manner. We examined the role of type I IFNs in macrophage responses to LPS using bone marrow-derived macrophages (BMM) from IFNAR1-/- mice, which lack a component of the type I IFN receptor and do not respond to type I IFNs. We found that, unlike wild-type (WT) BMM, LPS-treated IFNAR1-/- cells failed to produce nitric oxide (NO), or express inducible NO synthase (iNOS), indicating that type I IFNs are essential for all LPS-stimulated NO production in BMM. Exogenously added type II IFN (IFNgamma) rescued these responses in LPS-treated IFNAR1-/- BMM. In contrast to effects on NO, type I IFNs negatively regulated respiratory burst activity in LPS-primed BMM. We also found that while type I IFNs mediated the anti proliferative effects of lower concentrations of LPS, at higher concentrations LPS acted in a type I IFNs-independent manner. Finally, we report that type I IFNs are a survival factor for BMM. Despite this, the ability of LPS to also prevent apoptosis in BMM was independent of type I IFNs. These findings highlight the diverse roles of type I IFNs in mediating LPS-stimulated macrophage responses. PMID- 11052815 TI - Oncostatin M activates stat DNA binding and transcriptional activity in primary human fetal astrocytes: low- and high-passage cells have distinct patterns of stat activation. AB - In this study we explored the activation of the JAK/Stat pathway by gp 130 family cytokines in primary human astrocytes. We report that of four gp 130 cytokines tested, only oncostatin M (OnM) resulted in the activation of Stat molecules. To test that the induced molecules were transcriptionally active, transcription from a Stat-responsive reporter plasmid (from the acute-phase gene alpha-2 macroglobulin) transiently transfected into astrocytes was assessed after activation by OnM and was blocked by cotransfection with dominant-negative Stat3 encoding plasmids strongly suggesting that the activation was Stat-mediated. While DNA binding complexes comprised of both Stat1 and Stat3 were induced in low passage cells, only those containing Stat3 were formed by extracts from high passage cells. Stat1 protein was detected in the cytoplasm of high-passage cells indicating that the inability to form SIF-B and -C complexes was due to a lack of activation of Stat1 rather than a lack of expression. These results indicate a fundamental difference between low- and high-passage astrocytes in response to cytokine treatment that might result in distinct patterns of gene expression through altered ratios of activated Stat3 and Stat1. PMID- 11052816 TI - In vivo expression of the interleukin 4 receptor alpha by astrocytes in epilepsy cerebral cortex. AB - We reported previously that non-neoplastic astrocytes (derived from brain tissues of patients with epilepsy) expressed interleukin 4 receptor alpha (IL-4Ralpha) and responded to interleukin 4 (IL-4) in culture. To determine whether reactivity of cultured astrocytes was relevant to primary tissue, we investigated IL-4Ralpha expression in specimens of non-neoplastic cerebral cortex removed for surgical treatment of intractable epilepsy compared to specimens of glial tumours, which have been reported to contain IL-4Ralpha. Freshly frozen tissues from eight cases (four epilepsy, four malignant astrocytoma) were evaluated for IL-4Ralpha expression by reverse-transcriptase polymerase chain reaction (RT-PCR), Southern blotting, and double-labelled immunohistochemistry with antibodies to IL-4Ralpha and glial fibrillary acidic protein (GFAP). IL-4Ralpha mRNA was detectable in both non-neoplastic and neoplastic tissues, whereas interleukin 2 receptor gamma chain (IL-2Rgammac) mRNA was not found. By immunohistochemistry, IL-4Ralpha protein co-localized to cells displaying GFAP and astrocytic morphology in epilepsy tissues. As anticipated, IL-4Ralpha was detectable in astrocytoma, but, surprisingly, was also observed in GFAP-positive, non-neoplastic "reactive" astrocytes adjacent to tumour. Results are consistent with the concept that non neoplastic epilepsy astrocytes express IL-4Ralpha in situ, thus confirming in vitro studies and implying IL-4 sensitivity in vivo. PMID- 11052817 TI - Role of cytokine-induced neutrophil chemoattractant-2 (CINC-2) alpha in a rat model of chronic bronchopulmonary infections with Pseudomonas aeruginosa. AB - In order to investigate the role of the cytokine-induced neutrophil chemoattractant (CINC) in chronic bronchopulmonary infection, we developed a rat model of bronchopulmonary infection with Pseudomonas aeruginosa by using the agar bead method, and determined the kinetics of bacterial and cell number, as well as the concentrations of CINC-1, CINC-2, and CINC-3 in bronchoalveolar lavage (BAL) fluids in this model. The bacterial number in the lung rapidly increased from days 1 to 4, and declined 14 days after challenge. Neutrophil number in BAL fluid increased up to one day after challenge, and then slowly decreased during 14 days post-challenge. Among the CINCs, the local production of CINC-2 alpha sharply increased at day 1 and then decreased until day 4 post-challenge, while the local production of CINC-1 slightly increased at day 1 post-challenge. Neither CINC-2 beta nor CINC-3 were detected during the entire course of the infection. Increased CINC-2 mRNA expression in the lung tissue after challenge was associated with CINC-2 alpha production in BAL fluid. Moreover, an immunohistochemical study demonstrated the localization of CINC-1 and CINC-2 alpha primarily in alveolar macrophages and, to a much lesser extent, in bronchial epithelium of infected lung tissues, whereas CINC-2 beta and CINC-3 were not detected. When anti-CINC-1 or anti-CINC-2 alpha polyclonal antibodies were used for neutralizing neutrophil chemotactic activities in BAL fluids, the anti-CINC-2 alpha antibody inhibited 70% of the chemotactic activity in BAL fluids from infected rats at day 1 after challenge. No inhibition was observed by anti-CINC-1 antibody. These data indicate that CINC-2 alpha, which is produced by alveolar macrophages and bronchial epithelial cells, plays a pivotal role in neutrophil accumulation in the airway of a rat model of chronic bronchopulmonary infection with P. aeruginosa. PMID- 11052818 TI - The role of interleukin 6 in interferon-gamma production in thermally injured mice. AB - Following traumatic injury, patients suffer from compromised immunity increasing their susceptibility to infection. Previous studies from this laboratory demonstrated that female BALB/c mice subjected to a 15% total body surface area (TBSA) scald injury exhibit a decrease in cell-mediated immunity 10 days post burn. Studies described herein revealed that concanavalin A (Con A; 2 microg/ml) stimulated splenocytes from sham treated animals produced 3557+/-853 pg/ml of IFN gamma while splenocytes from burn injured animals released two-fold more cytokine (P<0.05). To determine whether leukocyte production of IFN-gamma was under the influence of macrophages, splenic macrophage supernatants generated from burned animals were incubated with splenic lymphocytes from sham and burn animals. The amount of IFN-gamma released by lymphocytes from sham animals increased when cultured with macrophages from burned mice (P<0.05). This suggests that the increase in IFN-gamma production by unfractionated splenocytes in burned mice relative to sham treated animals is macrophage-dependent. Macrophage supernatants from burned mice released twice as much IL-6 as supernatants from sham animals (P<0.05), and when IL-6 was blocked in vivo, the amount of IFN-gamma production in burned mice decreased to sham levels (P<0.05). Thus, IL-6 mediates IFN-gamma production following burn. PMID- 11052819 TI - Effects of leukaemia inhibitory factor on embryo implantation in the mouse. AB - Leukaemia inhibitory factor (LIF) is a pleiotrophic cytokine. Recent reports indicate that LIF is relevant to murine embryo implantation. In this work, results of indirect immunofluorescence under a confocal microscope illustrated that LIF was mainly located in the uterine lumen and uterine epithelial cells in pregnant mice on day 4. The number of embryos implanted in pregnant mice on day 8 decreased significantly after injection of 3 microg LIF antibodies into a uterine horn (P<0.001), which demonstrated again that LIF is a critical factor for embryo implantation. In a co-culture system, LIF (0.1 ng/ml, 1 ng/ml, 10 ng/ml and 100 ng/ml) significantly enhanced the blastocyst outgrowth after 24, 48 or 72 h of co culture, and outgrowth areas after 72 h of co-culture. Conversely, 5 microg/ml and 10 microg/ml, but not 1 microg/ml, LIF antibodies decreased the percentage of blastocysts with outgrowth; only 10 microg/ml LIF antibody inhibited blastocyst outgrowth area significantly (P<0.001). However, neither LIF nor its antibodies changed embryo attachment. Analysis of correlation showed that the effects of LIF or its antibodies on the blastocyst outgrowth were dose-dependent. In summary, different pathways may exist to regulate the blastocyst attachment and outgrowth on a monolayer of uterine epithelial cells. LIF protein from the maternal uterus exerts an essential role in embryo implantation in the mouse, which is mediated by stimulating trophoblast outgrowth, but not by promoting the attachment. PMID- 11052820 TI - Macrophage activation for the production of immunostimulatory cytokines by delivering interleukin 1 via biodegradable microspheres. AB - Interleukin 1alpha (IL-1alpha), a pleiotropic cytokine with multiple anti-tumour activities, has been investigated in our laboratory for its potential to serve as an immunotherapeutic agent. In the present study, an attempt was made to direct IL-1alpha to macrophages, in order to induce their immunoregulatory activities. For that purpose, IL-1alpha was encapsulated within biodegradable poly(lactic/glycolic acid) microspheres, 1-5 microm diameter in size. The microspheres were efficiently taken-up by macrophages in culture and after intraperitoneal injection into mice. In culture, phagocytosis of the microspheres reached saturation within 3 h and there was no apparent effect of polymer type on the extent of uptake. In vivo uptake of human IL-1alpha-microspheres by the macrophages lead to cell activation, as evidenced by the enhanced production of murine IL-1alpha, IL-6 and IL-12. Control microspheres, containing bovine serum albumin, induced only background to low levels of cytokine production. These cytokines, when expressed by or secreted from macrophages, may stimulate in situ diverse immune and inflammatory responses, including T cell-mediated immune responses, such as the development of Th(1)cells and cytotoxic lymphocytes. Thus, directing IL-1alpha into macrophages, via the appropriate microspheres, may serve as a unique mean to activate these cells to participate in anti-tumour immune responses in situ. PMID- 11052821 TI - Biopharmaceutics of liposomal interleukin 2, oncolipin. AB - Oncolipin is a multilamellar liposomal (dimyristoyl phosphatidylcholine) formulation of interleukin 2 (IL-2) and human serum albumin (HSA) with distinct surface characteristics which may influence its biological activities. IL-2 and HSA were detected on the surface of the liposomes using specific antibody staining. Surface expression of IL-2 was also demonstrated by the observation that Oncolipin bound to cells expressing IL-2 receptors (IL-2R) containing alphabetagamma or betagamma subunits. Binding and internalization of Oncolipin by cells expressing alphabetagamma or betagamma receptor subunits was blocked by excess free IL-2 or a neutralizing antibody against the beta chain. The display of surface IL-2 on Oncolipin's liposomes was maintained in vivo after intravenous injection into mice. IL-2 was also present between the lipid bilayers of the multilamellar liposomes based on the unique physical characteristics detected by freeze fracture electron microscopy. The bulk of the liposome-associated IL-2 was released from the liposomes upon incubation at 37 degrees C in medium containing serum, indicating that the IL-2 was not irreversibly entrapped on or in the liposome structure. Thus, Oncolipin is receptor-targeted to activated T and NK cells by virtue of its surface expression of IL-2 and has the potential to release IL-2 following deposition within lymphoid organs. These properties may confer distinct advantages over soluble IL-2 for immunotherapy of cancer and viral diseases. PMID- 11052822 TI - A comparison of biodistribution of liposomal and soluble IL-2 by a new method based on time-resolved fluorometry of europium. AB - A novel method was developed to determine the pharmacokinetics and biodistribution of cytokines and lymphokines based on time-resolved fluorometry (TRF) of europium (Eu). The comparison of two formulations of IL-2 was used to illustrate the sensitivity and applicability of this method as well as to extend the information on the pharmacokinetics of liposomal IL-2 and soluble IL-2. The blood kinetics and biodistribution of liposomal and soluble IL-2 in lymphoid organs and kidneys as measured by TRF were similar to those determined by the radioisotopic method. In both instances, the formulation of IL-2 into liposomes increased its serum half-life and accumulation in reticuloendothelial and lymphoid organs. The increased sensitivity of the Eu/TRF method permitted the extension of observational time points and the analysis of biodistribution in organs such as lymph nodes and bone marrow. These results suggest that Eu labelled proteins in conjunction with TRF offer a suitable alternative to radiolabelled proteins for pharmacokinetics and tissue distribution studies in animals. This method offers distinct advantages over traditional techniques employing radioistopes since it has greater sensitivity, no half-life limitations and no radioactive or hazardous waste disposal. PMID- 11052823 TI - Effects of sample handling on the stability of interleukin 6, tumour necrosis factor-alpha and leptin. AB - Detected levels of IL-6, TNF-alpha and leptin may be affected by methods of storage, anticoagulant or repeated freezing-thawing. Blood samples from 22 healthy subjects were: (i) allowed to stand for 1, 2, 4 or 6 h prior to, or after separation, before freezing at -70 degrees C; (ii) taken into tubes with lithium heparin, sodium citrate, EDTA or no anticoagulant, separated and frozen; and (iii) separated, and plasma repeatedly freeze-thawed for up to six cycles prior to assay. Leptin was assayed by RIA, and IL-6 and TNF-alpha by high-sensitivity ELISA. (i) IL-6 and TNF-alpha levels were not altered significantly in separated samples but IL-6 declined by mean (SEM) 14.3% (3.7%) and TNF-alphaincreased by 9.6% (2.3%) in samples left unseparated for 4 h (P=0.003 and 0.002, respectively). Leptin remained unchanged. (ii) Serum and EDTA-plasma samples gave comparable results for all three cytokines, but levels in the other anticoagulant samples were highly variable. (iii) IL-6 and leptin levels were not altered by up to 6 cycles of freeze-thawing, but TNF-alpha increased by 17.0% (3.7%) after 3 cycles. Concentrations of these molecules are significantly altered by storage conditions, therefore they need to be standardized for epidemiological and clinical studies, and between-study comparisons of levels may not be reliable. PMID- 11052824 TI - Prostaglandin E(2) and tumour necrosis factor-alpha release by monocytes are modulated by phospholipids. AB - The regulation of pro- and anti-mediator release from cells within the alveolar space would represent a desirable mechanism serving to protect this delicate gas exchanging region of the lung. This study investigates the effect of alveolar surfactant lipids on the regulation of tumour necrosis factor alpha (TNF-alpha), a potent inflammatory cytokine, and prostaglandin E(2)(PGE(2)), a lipid mediator with anti-inflammatory properties. The results of this investigation reveal a marked effect on the release of these two important mediators from a monocytic cell line, MonoMac 6 (MM6), by phosphatidylcholine (PC), phosphatidylethanolamine (PE), cholesterol (Chol) and sphingomyelin (SM). PC, PE and Chol demonstrated marked downregulation of TNF-alpha production at lipid concentrations of 125 and 250 microg/ml. Interestingly, SM significantly up regulated the release of TNF alpha at these concentrations. However, the release of PGE(2)in MM6 cells incubated with the same lipids was significantly increased with PC and Chol, and significantly decreased in cells pre-treated with SM. This indicates a role for these lipids in alveolar immunoregulation. PMID- 11052825 TI - Glutamine-induced cell swelling is not involved in the stimulatory effect of glutamine on cytokine production in rat peritoneal macrophages. PMID- 11052826 TI - Intrinsic capacity of monocytes to produce cytokines ex vivo in patients with acute lymphoblastic leukaemia. AB - Monocytic cytokine profiles of fifteen children with acute lymphoblastic leukaemia (ALL) were included to determine whether malignancy per se contributes to impaired cytokine profiles in vivo and ex vivo. The ex vivo tumour necrosis factor-alpha (TNF-alpha) and interleukin 1beta (IL-1beta) production was positively correlated with the monocyte number and with the number of intracellular TNF-alpha or IL-1beta positive cells in lipopolysaccharide (LPS) stimulated MNC cultures. The mean ex vivo TNF-alpha and IL-1beta production per 1x10(4)monocytes in these cultures was not significantly different in children at diagnosis of ALL, at remission or in controls. High IL-10 plasma levels at diagnosis of ALL had no effect on the ex vivo TNF-alpha and IL-1beta production of monocytes in LPS stimulated MNC cultures. These results show that monocytes of ALL patients have a normal intrinsic capacity to produce cytokines ex vivo. However, the decreased monocyte number is responsible for the lower TNF-alpha and IL-1beta concentrations ex vivo upon LPS stimulation. PMID- 11052827 TI - Modulation by soluble factors of gelatinase activities released by osteoblastic cells. AB - This study investigated the ability of normal human osteoblasts (hOb) and osteogenic sarcoma cells (MG-63 and SaOS2) to produce gelatinases and undergo modulation by interleukin 1beta (IL-1beta), interleukin 6 (IL-6), oncostatin M (OSM), leukaemia inhibitory factor (LIF), growth hormone (GH) and insulin-like growth factor-I (IGF-I). Gelatinase activities were determined by zymogaphy, and a quantitative analysis was performed by ELISA. The MMP-2 activities of the three cell lines were significantly increased in the presence of IL-1beta and IL-6, but no modulation of MMP-2 activities was observed in the presence of OSM, LIF and GH. IGF-I increased the activity released by SaOS2 and hOb, but no modulation was detectable in MG-63 cell conditioned medium. An upmodulation of pro-MMP-2 secretion by SaOS2 and hOb was observed for all soluble factors used, whereas an upmodulation of pro-MMP-2 secretion by MG-63 was observed only in the presence of IL-1beta, IL-6 and IGF-I. Thus, osteoblastic cells modulated by cytokines can be involved in bone resorption as a result of the protease activities released. PMID- 11052828 TI - TNF-alpha in the regulation of MUC5AC secretion: some aspects of cytokine-induced mucin hypersecretion on the in vitro model. AB - TNF-alpha has been implicated in the aetiology of otitis media with effusion (OME), where goblet cells proliferate in a modified respiratory epithelium, leading to the accumulation of a mucin-rich effusion in the middle-ear cleft. The MUC5AC mucin gene product has been identified as a component of these effusions. Here we have used the HT29-MTX goblet cell line, which secretes MUC5AC mucin, as a model to study the effect of TNF-alpha on goblet cells. MUC5AC mucin was identified and quantitated with a monoclonal antibody NCL-HGM-45M1. TNF-alpha stimulates MUC5AC mucin secretion in a dose-dependent manner, with 20 ng/ml producing maximal stimulation. Both pre-confluent and confluent cells showed peak stimulation after 7 h, however the pre-confluent cells showed twice the level of mucin hypersecretion. These results suggest that TNF-alpha stimulation of mucin secretion could play an important role in the early acute phase of the development of OME. This hypersecretion of mucin could then lead to the failure of the mucociliary clearance system, resulting in the accumulation of a mucin rich effusion in the middle ear and the movement to a more chronic phase of the disease. PMID- 11052829 TI - Effect of cytokines and colony-stimulating factors on passive polymorphonuclear leukocyte deformability in vitro. AB - Cytokines are potent polymorphonuclear leukocyte (PMN) activators and can decrease their deformability. We evaluated passive PMN deformability using the micropipette method after incubation with different concentrations of lipopolysaccharide (LPS), interleukins (IL-) 1, 6, 8 and 10, tumour necrosis factor (TNF), granulocyte (G) and granulocyte-macrophage (GM) colony-stimulating factors (CSF). TNF, IL-1, G-CSF, GM-CSF and, to a lesser degree, IL-6 significantly and in a dose-dependent fashion decrease PMN deformability. LPS had no direct effect on PMN deformability. When cytokines at concentrations with no effect on deformability were combined they increased PMN rigidity. The findings suggest that several cytokines and CSF impair directly, and not by activation alone, PMN deformability. PMID- 11052830 TI - Cardiotrophin-1 reduces stress-induced heat shock protein production in cardiac myocytes. AB - Cardiotrophin-1 (CT-1) can induce expression of the protective heat shock proteins (hsps) in cardiac cells. We show here that, unlike the stress induced accumulation of hsps, the effect of CT-1 is not accompanied by increased hsp mRNA levels and is insensitive to the RNA synthesis inhibitor actinomycin D, suggesting that it occurs at the post-transcriptional level. Pre-treatment with CT-1 reduces the ability of heat shock to induce hsp expression and this effect occurs at the transcriptional level. Hence, CT-1 and stress induce the hsps via different pathways which can antagonise one another. The mechanisms of these effects and their potential impact on the use of CT-1 as a cardioprotective agent are discussed. PMID- 11052831 TI - Coronary stent implantation: a panacea for the interventional cardiologist? PMID- 11052832 TI - Rotational atherectomy revisited in the era of stenting. PMID- 11052833 TI - Intracoronary drug delivery: mechanically too rough, pharmacologically too weak? PMID- 11052834 TI - Collateral flow and restenosis: appreciating hydraulics and outcomes of percutaneous coronary intervention. PMID- 11052835 TI - Small vessel stenting is safe, but still waiting for a well-proven antirestenotic effect. PMID- 11052836 TI - Blood glucose and coronary heart disease. PMID- 11052837 TI - Can the laser light at the end of the tunnel be used to get to the end of the tunnel? PMID- 11052838 TI - Coronary in-stent restenosis - predictors, treatment and prevention. PMID- 11052839 TI - Influence of diabetes mellitus on clinical outcomes across the spectrum of acute coronary syndromes. Findings from the GUSTO-IIb study. GUSTO IIb Investigators. AB - AIMS: We examined the characteristics, outcomes, and effects of hirudin vs heparin treatment of diabetic patients across the spectrum of acute coronary syndromes. METHODS AND RESULTS: We studied the 12,142 patients enrolled in the randomized GUSTO-IIb study. Diabetic patients (n=2175) were older, more often female, more often had prior cardiovascular disease, hypertension, and hyperlipidaemia, and less often were current smokers. Diabetic patients had a higher overall incidence of death or (re)infarction at 30 days (13.1% vs 8.5%, P=0.0001), whether they presented with ST-segment elevation (13.9% vs 9.9%, P=0.0017) or not (12.8% vs 7.8%, P=0.0001), and at 6 months (18.8% vs 11.4%, P=0.0001). Among diabetic patients, hirudin was associated with a tendency toward a lower risk of death or (re)infarction at 30 days (12.2% vs 13.9% with heparin) and 6 months (17.8% vs 20.2%). Diabetic patients had more major bleeding, stroke, heart failure, shock, atrioventricular block, and atrial arrhythmias, but no increased risk for ocular bleeding. CONCLUSIONS: Diabetic patients with acute coronary syndromes had worse 30-day and 6-month outcomes, particularly those without ST-segment elevation. The statistically non-significant trend toward improved outcomes with hirudin was similar among patients with and without diabetes, with a greater point estimate for the absolute difference in patients with diabetes. PMID- 11052840 TI - A randomized comparison of balloon angioplasty versus rotational atherectomy in complex coronary lesions (COBRA study). AB - AIMS: Rotablation is a widely used technique for the treatment of complex coronary artery lesions but is so far only poorly supported by controlled studies. The Comparison of Balloon-Angioplasty versus Rotational Atherectomy study (COBRA) is a multicentre, prospective, randomized trial to compare short- and long-term effects of percutaneous transluminal coronary angioplasty (PTCA) and rotablation in patients with angiographically pre-defined complex coronary artery lesions. METHODS AND RESULTS: At seven clinical sites 502 patients with pre-defined complex coronary artery lesions were assigned to either PTCA (n=250) or rotablation (n=252). Primary end-points were procedural success, 6-month restenosis rates in the treated segments, and major cardiac events during follow up. Procedural success was achieved in 78% (PTCA), and 85% (rotablation) (P=0.038) of cases. Crossover from PTCA to rotablation was 4% and 10% vice versa (P=0.019). There was no difference between PTCA and rotablation with respect to procedure-related complications such as Q wave infarctions (2.4% each), emergency bypass surgery (1.2% versus 2.4%), and death (1.6% versus 0.4%). However, more stents were required after PTCA (14.9% versus 6.4%, P<0.002), predominantly for bailout or unsatisfactory results. Including bail-out stents as an end-point, the procedural success rates were 73% for angioplasty and 84% for rotablation (P=0.006). At 6 months, symptomatic outcome, target vessel reinterventions and restenosis rates (PTCA 51% versus rotablation 49%, P=0.33) were not different. CONCLUSION: Complex coronary artery lesions can be treated with a high level of success and low complication rates either by PTCA with adjunctive stenting or rotablation. The long-term clinical and angiographic outcome is comparable. PMID- 11052841 TI - Local delivery of nadroparin for the prevention of neointimal hyperplasia following stent implantation: results of the IMPRESS trial. A multicentre, randomized, clinical, angiographic and intravascular ultrasound study. AB - BACKGROUND: We hypothesized that intramural delivery of nadroparin, a low molecular weight heparin, would prevent in-stent restenosis by inhibiting neointimal hyperplasia in an angioplasty model free of arterial remodelling. METHODS AND RESULTS: In a prospective randomized multicentre trial, 250 patients submitted to balloon angioplasty followed by stent implantation were randomized into control group (no local drug delivery) or intramural delivery of nadroparin (2 ml of 2500 anti-Xa-units/ml with a microporous catheter). An ancillary intravascular ultrasound substudy was performed to supplement angiographic data with specific measurements of in-stent neointimal hyperplasia. The primary end point was the late loss in minimal luminal diameter on the 6 month follow-up angiogram. Secondary end-points included feasibility and safety of local nadroparin delivery, and major adverse cardiac events at 8 weeks and 6 months follow-up. Local delivery of nadroparin was successful in 124 patients (99.2% success rate) and was not associated with an increase in stent thrombosis, coronary artery dissection, side branch occlusion, distal embolization or abrupt arterial closure. At angiographic follow-up, the late loss in lumen diameter was 0.84 +/- 0.62 mm in the control group compared to 0.88 +/- 0.63 mm in the nadroparin group (P=0.56). Angiographic restenosis rate (defined as a >50% diameter stenosis) did not differ in the control group (20%) compared to the nadroparin group (24%). The average area of neointimal tissue within the stent was 2.86 +/- 0.64 mm(2) vs 2.90 +/- 0.53 mm(2) (P=0.57), control vs nadroparin groups. There was no difference in major adverse cardiac events at any time (88.8% vs 89.6% event free survival at 6 months, control vs nadroparin). CONCLUSION: Intramural delivery of nadroparin with a microporous catheter after stent deployment was feasible and safe but had no effect in reducing restenosis or the occurrence of major adverse clinical events over 6 months. PMID- 11052842 TI - Quantitatively assessed coronary collateral circulation and restenosis following percutaneous revascularization. AB - AIMS: A high degree of collateral supply to a vascular area where a percutaneous transluminal coronary angioplasty (PTCA) has been performed represents a haemodynamic force competing with the antegrade flow through the dilated lesion. Therefore, our purpose was to determine whether patients with restenosis following PTCA have a higher collateral flow to the recipient vessel than patients without restenosis. METHODS AND RESULTS: In 200 consecutive PTCA patients, an intracoronary pressure-derived collateral flow index (CFI) was determined quantitatively during balloon occlusion, using simultaneous measurements of the mean aortic pressure (P(ao)) and of the intracoronary pressure distal to the occluded stenosis (P(occl)), as well as the estimated central venous pressure (CVP=5 mmHg): CFI=(P(occl)-CVP)/(P(ao)-CVP). Sixty-four patients had an angiographic follow-up examination after at least 2 months, and were subdivided into patients with restenosis (>50% diameter stenosis, n=34) and patients without restenosis (n=30). Patients with restenosis had a significantly higher collateral flow index at the initial coronary angiography than patients without restenosis (0.26 +/- 0.14 vs 0.12 +/- 0.09; P<0.0001). CONCLUSIONS: Patients with restenosis after PTCA show a more extended collateral supply to this recipient area than patients without restenosis. Well developed collaterals to a revascularized region are a risk factor for restenosis of the treated lesion. PMID- 11052843 TI - Randomized comparison of coronary stenting with optimal balloon angioplasty for treatment of lesions in small coronary arteries. AB - AIMS: Angioplasty of lesions in small coronary arteries remains a significant problem because of the increased risk of restenosis. The aim of this study was to compare the efficacy of elective coronary stent placement and optimal balloon angioplasty in small vessel disease. METHODS: One hundred and twenty patients with lesions in small coronary arteries (de novo, non-ostial lesion and reference diameter <3 mm) were randomly assigned to either balloon angioplasty or elective stent placement (7-cell NIR stent). The primary end-point was restenosis at 6 months follow-up. Optimal balloon angioplasty was defined as diameter stenosis less than or = 30% and the absence of major dissection after the angioplasty, and crossover to stenting was allowed. RESULTS: Baseline clinical and angiographic characteristics were similar in the two groups. Procedure was successful in all patients, and in-hospital events did not occur in any patient. However, 12 patients in the angioplasty group were stented because of suboptimal results or major dissection. Postprocedural lumen diameter was significantly larger in the stent group than in the angioplasty group (2.44 +/- 0.36 mm vs 2.14 +/- 0.36, P<0.05, respectively), but late loss was greater in the stent group (1.12 +/- 0.67 mm vs 0.63 +/- 0.48, P<0.01, respectively). The angiographic restenosis rate was 30.9% in the angioplasty group, and 35.7% in the stent group (P = ns). Clinical follow-up was available in all patients (15.9 +/- 5.7 months) and clinical events during the follow-up were similar in both groups. CONCLUSIONS: These results suggest that optimal balloon angioplasty with provisional stenting may be a reasonable approach for treatment of lesions in small coronary arteries. PMID- 11052844 TI - Abnormal glucose tolerance, not small vessel diameter, is a determinant of long term prognosis in patients treated with balloon coronary angioplasty. AB - AIMS: We sought to find out what factors are important for long-term prognosis, the small vessel itself or abnormal glucose tolerance, in patients treated with coronary angioplasty. BACKGROUND: Patients with coronary artery disease with diabetes mellitus often show diffuse and small coronary artery narrowing. Impaired glucose tolerance has also been reported to be a risk factor for cardiovascular disease. METHODS: Among 584 patients who underwent first elective balloon coronary angioplasty, diabetes mellitus and impaired glucose tolerance were present in 197 patients. Large and small vessels were defined by reference vessel diameter before coronary angioplasty as either larger or smaller than 2.5 mm. Patients were categorized into the following four groups: 175 patients with normal glucose tolerance and reference diameter <2.5 mm (group SN), 212 patients with normal glucose tolerance and reference diameter greater than or = 2.5 mm (group LN), 101 patients with abnormal glucose tolerance and reference diameter <2.5 mm (group SD), and 96 patients with abnormal glucose tolerance and reference diameter greater than or = 2.5 mm (Group LD). The cardiac events were compared for a period of 8 years after coronary angioplasty among the four groups. RESULTS: There was no difference in the percentage diameter stenosis immediately after coronary angioplasty among the four groups. However, group SD showed unfavourable prognosis despite similar minimal lumen diameter after coronary angioplasty compared with group SN. Event-free survival curve of group LD showed a sudden drop approximately 5 years after the coronary angioplasty. In multivariate analysis, the cardiac events were associated with the presence or absence of abnormal glucose tolerance. Furthermore, patients with bad glycaemic control (HbA1c>6.0%) at index coronary angioplasty showed worse event free survival than those with good glycaemic control. CONCLUSIONS: An important determinant for long-term prognosis after coronary angioplasty is a presence of abnormal glucose tolerance per se and not small vessel diameter. PMID- 11052845 TI - Total occlusion trial with angioplasty by using laser guidewire. The TOTAL trial. AB - AIMS: A randomized trial was performed to assess the safety and efficacy of a laser guidewire, in the treatment of chronic coronary occlusions. METHODS AND RESULTS: In 18 European centres, 303 patients with a chronic coronary occlusion were randomized to treatment with either the laser guidewire (n=144) or conventional guidewires (mechanical guidewire, n=159). The primary end-point of the study was treatment success, defined as reaching the true lumen distal to the occlusion by the allocated wire within 30 min of fluoroscopic time: laser guidewire vs mechanical guidewire; 52.8% (n=76) vs 47.2% (n=75), P=0.33. Serious adverse events following the initial guidewire attempt were 0% (laser guidewire) and 0.6% (mechanical guidewire), respectively. Angioplasty (performed following successful guidewire crossing) was successful in 179 patients (91%, laser guidewire n=79, mechanical guidewire n=100), followed by stent implantation in 149 (79%). At the 6-month angiographic follow-up, the difference in binary restenosis rate (laser guidewire vs mechanical guidewire; 45.5% vs 38.3 %, P=0.72) or reocclusion rate (25.8% vs 16.1%, P=0.15) did not reach statistical significance. At 1, 6 and 12 months, angina and event-free survival were 69%, 35% and 24% (laser guidewire) vs 74%, 40% and 31% (mechanical guidewire). CONCLUSION: Although laser guidewire technology was safe, the increase in crossing success did not reach statistical significance. PMID- 11052846 TI - ESC news and appointments. ProCOR: using information technology to address cardiovascular diseases in developing countries. PMID- 11052847 TI - Home-based intervention: the next step in treatment of chronic heart failure? PMID- 11052848 TI - Genes and acquired disease: beta-adrenoceptor polymorphisms and heart failure. PMID- 11052849 TI - Semicarbazide-sensitive amine oxidase and mortality in chronic heart failure. PMID- 11052850 TI - Prognostic stratification in heart failure: what's the point? PMID- 11052851 TI - Genetics and dilated cardiomyopathy: limitations of candidate gene strategies. PMID- 11052852 TI - Parallel realities of guidelines and practice. PMID- 11052853 TI - The Tei index - a role in the diagnosis of heart failure? PMID- 11052854 TI - Familial dilated cardiomyopathy: from clinical presentation to molecular genetics. PMID- 11052855 TI - Trends in case-fatality in 117 718 patients admitted with acute myocardial infarction in Scotland. AB - OBJECTIVES: To analyse short- and long-term case-fatality trends following admission to hospital with a first acute myocardial infarction, in men and women between 1986 and 1995, after adjusting for risk factors known to influence survival. DESIGN: A Scottish-wide retrospective cohort study. SETTING: The Linked Scottish Morbidity Record Database was analysed. This contains accurate data on all hospital admissions since 1981, for the Scottish population of 5.1 million. It is linked to the Registrar General's death certificate data. SUBJECTS: All 117 718 patients admitted to Scottish hospitals with a principal diagnosis of first acute myocardial infarction (ICD-9 code 410) between 1986 and 1995. MAIN OUTCOME MEASURES: The outcome was death, both in and out of hospital, from any cause, at 30 days, 1 year, 5 and 10 years. RESULTS: Overall case-fatality following hospital admission with acute myocardial infarction was 22. 2%, 31.4%, 51.1% and 64.0% at 1 month, 1 year, 5 and 10 years, respectively. Multivariate analyses identified statistically significant independent prognostic factors. Thirty day mortality increased twofold for each decade of increasing age, and increased with any prior admission to hospital. When comparing the most deprived category to that of the most affluent, men had a 10% increased mortality (P<0.01), whilst women had an increased mortality of 4% (not significant). After adjustment for age, sex, deprivation and prior admission to hospital, case-fatality rates fell significantly between 1986 and 1995. Short-term case-fatality fell by 46% in men (27% in women) and long-term by 34% in men (30% in women) (both P<0.001). CONCLUSIONS: Population-based case-fatality rates in Scotland have fallen dramatically since 1986, particularly in men. The increasing survival in patients admitted to hospital suggests that the trial-based efficacy of modern therapies is now translating into population-based effectiveness. However, an individual's life expectancy still halves after a diagnosis of acute myocardial infarction. Of the variables that we could examine, age was the most powerful predictor of prognosis. PMID- 11052856 TI - The association between mortality from ischaemic heart disease and mortality from leading chronic diseases. AB - AIMS: Coronary risk factors raise the risk of other chronic disorders. We therefore tested the hypothesis that the geographic distribution of ischaemic heart disease mortality is associated with that of other chronic diseases with which it shares risk factors. METHODS AND RESULTS: For the 50 provinces of Spain, we collected mortality data for the period 1980-1995 from the national vital statistics. We calculated age-adjusted mortality rates for the leading causes of death in quintiles of provincial distribution of ischaemic heart disease mortality, and correlation coefficients with respect to provincial ischaemic heart disease mortality. As expected, because they share risk factors with ischaemic heart disease, mortality from cerebrovascular disease, malignant tumours, lung cancer, respiratory diseases, chronic obstructive pulmonary disease, diseases of the digestive system, cirrhosis of the liver and all causes, increase with the rise from lower to higher quintiles of ischaemic heart disease mortality. Ischaemic heart disease mortality registered correlations over 0.5 (P<0.001) with mortality from many of the above diseases in the periods 1980-1984 and 1991-1995. Expectations were similarly borne out for disorders not sharing risk factors with ischaemic heart disease, in that mortality from prostate and breast cancer, injury and poisoning, traffic accidents and ill-defined causes in most cases did not show a provincial association with ischaemic heart disease mortality. In general, these results were observed for both sexes and across all age groups. CONCLUSION: Ischaemic heart disease mortality is associated with mortality from chronic diseases which share coronary risk factors, across provinces of Spain over the period 1980-1995. This suggests that the geographic variation in such chronic diseases is due to common factors, potentially susceptible to similar preventive interventions. PMID- 11052857 TI - A novel polymorphism in the gene coding for the beta(1)-adrenergic receptor associated with survival in patients with heart failure. AB - AIMS: The adrenergic nervous system is of major importance in congestive heart failure. No genetic polymorphism has previously been identified in the beta(1) adrenergic receptor gene. The aim of this study was to find possible mutations in this gene and to relate such findings to morbidity and prognosis in heart failure. METHODS AND RESULTS: Genomic DNA was extracted from blood leukocytes from patients with congestive heart failure (n=184) and from age-matched controls (n=77). The part of the beta(1)-adrenergic receptor gene corresponding to nucleotide 1-255 was amplified by polymerase chain reaction and analysed by automated sequencing. The patients were investigated by echocardiography and followed regarding symptoms and survival for 5 years. A missense mutation was identified at nucleotide position 145 in the beta(1)-adrenergic receptor gene, which predicted an amino acid substitution at position 49 (Ser49Gly). The allele frequency of the Gly49 variant was 0.13 in controls and 0.18 in patients (P=0.19). At the time of the 5-years follow-up, 62% of the patients with the wild type gene and 39% of the patients with the Ser49Gly variant had died or had experienced hospitalization (P=0.005). Patients without the mutation had significantly poorer survival compared to those with the mutation, risk ratio 2.34 (95% CI 1.30-4.20), P=0.003. In a mulivariate analysis, the risk ratio was 2.03 (95% CI 0.99-4.16) P=0.05. CONCLUSION: A novel missense mution in the beta(1)-adrenergic receptor gene was associated with a decreased mortality risk in patients with congestive heart failure. These data suggest that the beta(1) receptor Ser49Gly variant might be associated with altered receptor function, resulting in myocardial protection in patients with heart failure. PMID- 11052858 TI - Plasma semicarbazide-sensitive amine oxidase (SSAO) is an independent prognostic marker for mortality in chronic heart failure. AB - AIMS: Experimental evidence has suggested that semicarbazide-sensitive amine oxidase is involved in vascular endothelial damage and in the process of atherosclerosis, through the formation of reactive aldehydes, hydrogen peroxide and ammonia from endogenous substrates. Recent evidence indicates that semicarbazide-sensitive amine oxidase may be identical with the vascular adhesion protein-1. In patients with diabetes mellitus and chronic heart failure the plasma activity is raised relative to the severity of the disease. The prognostic value of plasma semicarbazide-sensitive amine oxidase is not known. METHODS AND RESULTS: Plasma semicarbazide-sensitive amine oxidase activity was measured at baseline in patients with moderate to severe chronic heart failure who participated in a large European study (PRIME-II). The 372 patients who took part in a pre-defined substudy in The Netherlands were investigated and a survival follow-up (maximum 5.4 years, mean 3.4 years) was carried out. Within the follow up period 195 patients died. Plasma semicarbazide-sensitive amine oxidase was higher at baseline in those who died than in the survivors (653+/-258 vs 540+/ 242 mU. l(-1), P<0.001). Dividing the patients into two groups according to plasma values above or below the median value of 550 mU. l(-1), semicarbazide sensitive amine oxidase was found to be a prognostic parameter for survival, both in univariate (P<0.0001) and in multivariate (P=0.0106) analysis. Semicarbazide sensitive amine oxidase values >550 mU. l(-1)had a 1. 50 (95% CI, 1.10-2.04) times increased risk of death. CONCLUSION: The finding that plasma semicarbazide sensitive amine oxidase is an independent prognostic marker for mortality in chronic heart failure supports the concept that an elevated plasma semicarbazide sensitive amine oxidase level has deleterious effects, possibly due to vascular endothelial damage. PMID- 11052859 TI - Comparison of the prognostic value of left ventricular filling and peak oxygen uptake in patients with systolic heart failure. AB - AIM: The aim of this prospective study was to compare the prognostic value of the mitral inflow pattern and peak oxygen uptake in patients with systolic heart failure. BACKGROUND: Peak oxygen uptake is a major prognostic parameter in heart failure. It is not known whether a restrictive mitral inflow pattern has similar prognostic value. METHODS: One hundred heart failure patients (ejection fraction <45%) underwent exercise testing after Doppler evaluation; prognosis was assessed after a mean follow-up of 17 months. RESULTS: The ejection fraction was larger in group 1 (non-restrictive pattern: E/A mitral wave ratio <1 or between 1 and 2 with E wave deceleration time >/=140 ms, n=45) than in group 2 (restrictive pattern: E/A ratio >2 or between 1 and 2 with E deceleration time <140 ms, n=40) (29+/-9 vs 22+/-10%, P<0.05). Peak oxygen uptake was lower in group 2 (17+/-4 vs 22+/-5 ml. min(-1). kg(-1)57+/-11 vs 75+/-15% of predicted values;P<0.05 for both comparisons). Univariate analysis showed that the deceleration time (r=0.65), E/A ratio (r=-0.50) and heart rate increment (r=0.47) correlated best with peak oxygen uptake. A third group of patients with persistent fusion of the E and A waves (n=15) had exercise responses similar to those of group 2 patients. A short deceleration time (P=0.006), a restrictive or a fusion pattern (P=0.04) were associated with a poor outcome; the prognostic value of these Doppler variables was greater than that of ejection fraction, but remained less than peak oxygen uptake indexed by predicted values (P=0.0004). CONCLUSION: The left ventricular filling pattern is a strong predictor of exercise capacity, and outcome, in patients with systolic heart failure and is independent of the left ventricular ejection fraction. Peak oxygen uptake remains a more powerful prognostic variable. PMID- 11052860 TI - Epidemiology of desmin and cardiac actin gene mutations in a european population of dilated cardiomyopathy. AB - AIMS: Although dilated cardiomyopathy is the most frequent form of cardiomyopathy, its aetiology is still poorly understood. In about 20-30% of cases the disease is familial with a large predominance of autosomal dominant transmission. Ten different chromosomal loci have been described for autosomal dominant forms of dilated cardiomyopathy. Only two genes have been associated with pure forms (without myopathy and/or conduction disorders) of the disease, the cardiac actin and the desmin genes. Our aim was to determine the proportion of dilated cardiomyopathy affected individuals carrying a mutation in one of these two genes. METHODS AND RESULTS: We performed (1) a systematic polymerase chain reaction-SSCP-sequencing screening of the coding sequences of cardiac actin on DNA samples from 43 probands of dilated cardiomyopathy families and 43 sporadic cases; (2) a systematic polymerase chain reaction-SSCP-sequencing screening of the coding sequences of desmin combined with a search for the described missense mutation (Ile451Met) by restriction fragment length polymorphism analysis on DNA samples from 41 probands of dilated cardiomyopathy families and 22 sporadic cases. CONCLUSION: None of the patients presents a mutation in any of these two genes. Consequently, the proportion of European dilated cardiomyopathy affected individuals bearing a mutation in (1) the cardiac actin gene is less than 1.2%, (2) the desmin gene is less than 1.6%. PMID- 11052861 TI - European survey of primary care physician perceptions on heart failure diagnosis and management (Euro-HF). AB - AIMS: To survey a random sample of primary care physicians across six European countries regarding their perceptions of diagnostic and prescribing issues in heart failure, and to consider factors that might be associated with physician under-performance. METHODS AND RESULTS: Qualitative, postal questionnaire-based, validated survey in the native tongue of a random sample of 200 primary care physicians in each of five European countries (France, Germany, Italy, The Netherlands and Spain) and of 250 U.K. primary care physicians. Respondents provided: details of practice characteristics; the usual way a diagnosis of heart failure was established; access to investigations; names of drugs prescribed in heart failure, with estimates of the proportion of patients supplied with particular classes; and physician attitudes regarding the evidence base (in terms of benefits and risks) for treatments used. Outcomes were physician perceptions and attitudes about heart failure diagnosis and treatment. Adjusted response rates varied from 17% (France) to 56% (Britain). Primary care physicians underestimate the prevalence of heart failure. Most patients are diagnosed on symptoms and signs alone, with only 32% having further investigations or referral. Although most primary care physicians stated they prescribe ACE inhibitors in heart failure, this was for only 47-62% of patients, and at doses below those identified as effective in trials. Most prescribing doctors (91%) believe there is strong evidence of reduced mortality in heart failure patients using ACE inhibitors, but 51% also consider ACE inhibitors have substantial risks with their use. CONCLUSION: Limitations of the data include the general problem of questionnaires, whether responses accord with actual clinical practice, and, specific to these data, the low response rate in some countries (although the study does provide information from nearly 300 randomly selected primary care physicians across Europe). New preliminary insights include exposition of the 'low tech' approach to heart failure diagnosis across Europe: doctors report the use of symptoms and signs alone; the lack of direct (open) access to objective investigations, such as echocardiography, which almost guarantees that misdiagnoses will occur; and the under-utilization and under-dosing with ACE inhibitors. The main factor influencing under-use would appear to be the exaggerated perceptions of treatment risk amongst primary care physicians that dominate the widespread and accurate knowledge of treatment benefits. PMID- 11052862 TI - Tei-index in patients with mild-to-moderate congestive heart failure. AB - BACKGROUND: Congestive heart failure is related to contraction and relaxation abnormalities of the ventricle. Isolated analysis of either mechanism may not be reflective of overall cardiac dysfunction. A combined myocardial performance index (isovolumic contraction time plus isovolumic relaxation time divided by ejection time, 'Tei-Index') has been described which may be more effective for analysis of global cardiac dysfunction than systolic and diastolic measures alone. It was the aim of the present investigation to evaluate the Tei-Index against invasive examination. METHODS AND RESULTS: Eighty-one subjects were included in a consecutive manner, among 125 patients undergoing left heart catheterization for invasive measurement of left ventricular end-diastolic pressure; 43 patients had congestive heart failure (35 male, 8 female, 68+/-6 years) defined by NYHA functional class >/=2 (mean 2.5+/-0.5) and left ventricular end-diastolic pressure >/=16 mmHg (mean 20+/-4) and 38 subjects (32 male, 6 female, 66+/-5 years) without symptoms of heart failure (NYHA functional class I) and with normal left ventricular end-diastolic pressure (mean 12+/-3 mmHg) served as a control group. Using conventional echo-Doppler methods, parameters assessed were: ejection fraction, peak velocities of early (E) and late (A) diastolic filling, the E/A ratio, deceleration time, isovolumic contraction time, isovolumic relaxation time and ejection time. The Tei-Index was obtained by subtracting ejection time from the interval between cessation and onset of the mitral flow. The control group and patients with congestive heart failure did not differ with respect to the E/A ratio (0.86+/-0.27 vs 0.90+/-0.44, P=ns), deceleration time (203+/-42 ms vs 206+/-36 ms, P=ns) and isovolumic relaxation time (97+/-16 ms vs 94+/-26 ms, P=ns). The ejection fraction was slightly reduced in patients with congestive heart failure (46+/-11% vs 55+/-8%, P<0.05). The Tei-Index was easily and reproducibly measured in all subjects. The mean value of the Tei-Index was significantly different between the control group and patients with congestive heart failure (0.39+/-0.10 vs 0.60+/-0.18, P<0.001). Receiver operating characteristic curve analysis for the Tei-Index yielded an area under the curve of 0.88+/-0.038. Using a Tei-Index >/=0.47 as the cutpoint, congestive heart failure was identified with a sensitivity of 86% and a specificity of 82%. No correlation was observed between the Tei-Index and heart rate (r=0.22, P=ns), systolic blood pressure (r=0.16, P=ns) or diastolic blood pressure (r=0.08, P=ns). The Tei-Index was significantly related to left ventricular end-diastolic pressure (r=0.46, P<0.01). CONCLUSION: The Tei-Index is a sensitive indicator of overall cardiac dysfunction in patients with mild-to moderate congestive heart failure. The Tei-Index is easily obtained and may be used in the work-up of patients with suspected cardiac dysfunction. PMID- 11052863 TI - ESC news and appointments PMID- 11052864 TI - Trypanosoma cruzi: infectivity of clonal genotype infections in acute and chronic phases in mice. AB - Eight Trypanosoma cruzi stocks pertaining to the clonal genotypes 19/20, 32, and 39 have been characterized for three experimental parameters of infectivity in Balb/c mice: (i) percentage of mice with a patent parasitemia (% MPP), (ii) maximum parasitemia (MP), and (iii) percentage of mice with positive hemoculture (% MPH). By order of decreasing values, the values recorded for the clonal genotypes ranked as follows: 19/20, 32, and 39, except for the % MPP parameter, for which 19/20 and 32 were not statistically different. The rate of successful reisolation after infection in mice, analyzed by multilocus enzyme electrophoresis and random amplified polymorphic DNA typing, was statistically different according to the clonal genotype and was different for uniclonal infections and for mixed infections by two different clonal genotypes. These results confirm that T. cruzi clonal genotypes differ significantly in their infectivity in mice. PMID- 11052865 TI - Trypanosoma brucei: identification of trypanosomes with genotypic similarity to human infective isolates in tsetse isolated from a region free of human sleeping sickness. AB - In previous work, we have developed a molecular method that defines genotypes of Trypanosoma brucei and allows distinction of the human-infective subspecies T. b. rhodesiense from the non-human-infective T. b. brucei without recourse to measurement of resistance to lysis by human serum. Using this approach, we are also able to determine the geographical range of specific genotypes associated with a particular focus. In this study, we have characterised T. brucei isolates collected from tsetse in a region where human sleeping sickness has never been reported and which is some 500 km from the Busoga sleeping sickness focus of Uganda. We show that some of the trypanosome isolates taken from tsetse in this region have considerable genotypic similarity to trypanosomes from the Busoga focus, demonstrating a surprisingly wide dispersal of these trypanosome genotypes. Furthermore, the similarity of these genotypes to human-infective trypanosomes in the Busoga focus suggest the possible circulation of human infective trypanosomes in this location. We also demonstrate that the genetic diversity in trypanosomes isolated from tsetse is significantly higher than that in those isolated from humans, confirming other studies that show that there exists a significant restriction in the range of genotypes that can be transmitted to humans. PMID- 11052866 TI - Role of nonspecific cytotoxic cells in the induction of programmed cell death of pathogenic protozoans: participation of the Fas ligand-Fas receptor system. AB - Numerous different species of parasites and pathogenic microorganisms produce programmed cell death (PCD) and apoptosis in eukaryotic targets. How ever, only a few studies have demonstrated that effector cells, cytokines, growth factors, or soluble apoptosis-inducing factors are capable of initiating apoptosis in protozoan parasites. Certain Tetrahymena spp. in teleosts are opportunistic pathogens. In the present study these pathogenic protozoans were developed as a model system to describe the potential role of the Fas ligand (FasL)-Fas receptor (FasR) system as a means of innate immunity in teleosts. Nonspecific cytotoxic cells (NCC) constitutively express soluble FasL (sFasL). Binding of the antigen receptor (i.e., NCCRP-1) on NCC to target cells caused the release of sFasL into the milieu. The presence of functional sFasL in these supernatants was determined by Western blot analysis and by demonstrating the lysis of FasR(+) HL-60 but not IM-9 (FasR(-)) targets. Soluble FasL containing supernatants generated by tumor cell-activated NCC also produced a reduction in 2 N DNA (i.e., DNA hypoploidy) of T. furgasoni. The induction of DNA hypoploidy by NCC supernatants could be neutralized by adsorption of the supernatants with anti-FasL antibody (but not with an isotype control). Experiments were next done to determine the expression of FasR on Tetrahymena and study the effects of anti-FasR monoclonal crosslinkage and treatment with soluble human recombinant FasL (huFasL) on initiation of PCD in Tetrahymena. Cell cycle analysis revealed that both crosslinkage and soluble huFasL binding to Tetrahymena produced DNA hypoploidy. The reduction in diploid DNA was confirmed by observing oligonucleosome fragmentation (DNA laddering) following anti-FasR treatment. Additional evidence for FasR expression on Tetrahymena was obtained using fluorescence microscopy and flow cytometry. Both methods showed that all Tetrahymena examined (three species consisting of four isolates) expressed membrane FasR. These studies demonstrated the potential of the FasL-FasR system in teleosts for initiation of antiparasite innate immunity. Effector NCC may initiate PCD of Tetrahymena that express a FasR-like protein. Induction of apoptosis may be a major mechanism of homeostatic control of protozoan parasite infestations/infections. PMID- 11052867 TI - Toxoplasma gondii: identification of a developmentally regulated family of genes related to SAG2. AB - Previous studies have shown the surface of Toxoplasma gondii to be dominated by a family of proteins closely related to SAG1. In this study, we report the existence of a second family of genes defined by homology to SAG2. The predicted amino acid sequences of these three new proteins suggests that they are all glycosylphosphatidylinositol-linked surface antigens. All three also contain N linked glycosylation sites, although their use has yet to be demonstrated. One of these SAG2-related antigens, SAG2B, is expressed in tachyzoites with an apparent size of 23 kDa. It is distinct, however, from the previously identified P23. In contrast to SAG2B, SAG2C and SAG2D appear to be expressed exclusively on the surface of bradyzoites. Analysis of the SAG2 family shows it to have weak but significant homology to the SAG1 family. Thus, all of the sequenced surface antigens of tachyzoites and many of those of bradyzoites fall into one large superfamily that can be divided into two subgroups defined by the prototypic and highly immunogenic SAG1 and SAG2, respectively. PMID- 11052868 TI - Trypanosoma cruzi: acute infection affects expression of alpha-2-macroglobulin and A2MR/LRP receptor differently in C3H and C57BL/6 mice. AB - Although a complete cellular and humoral immune response is elicited in Chagas' disease, recent data suggest that other natural elements of innate immunity may also contribute to the initial host primary defense. alpha-Macroglobulins are a family of plasma proteinase inhibitors that are acute-phase reactants in Trypanosoma cruzi-infected mice and humans. Mice contain a tetrameric alpha-2 macroglobulin (MAM) and a monomeric murinoglobulin (MUG). Heterogeneity in their reactions was observed in murine T. cruzi-infected plasma A2M levels despite an overall increase. In addition, up-regulation of the A2M receptor (A2MR/LRP) was observed in peritoneal macrophages during T. cruzi infection. Here, we show that during T. cruzi infection (Y strain), the MAM and MUG hepatic mRNA levels and the corresponding plasma protein levels were up-regulated in C3H and C57BL/6 (B6) mice, but with different kinetics. On the contrary, A2MR/LRP mRNA levels increased in acutely infected C3H mice, but decreased in B6 mice, in both liver and heart. Immunocytochemistry of infected B6 heart cryosections confirmed a less intense endothelium labeling by the fluoresceinated ligand for A2MR/LRP. On the other hand, infected B6 spleen cells displayed higher F-A2M-FITC binding and MAC1 expression, confirming higher A2MR/LRP expression in macrophages. In uninfected mice, as well as after T. cruzi infection, higher A2M plasma levels were measured in C3H mice than in B6 mice. The lower tissue T. cruzi parasitism found in C3H infected mice could reflect an inhibitory effect of A2M on parasite invasion. Our present data further contribute to clarifying aspects of the role of A2MR/LRP in a model of acute Chagas' disease in different mouse strains. PMID- 11052869 TI - Leishmania infantum: gene cloning of the GRP94 homologue, its expression as recombinant protein, and analysis of antigenicity. AB - The complete nucleotide sequence for the Leishmania infantum homologue to the glucose-regulated protein 94 (GRP94) gene was determined from the isolation and characterization of a genomic clone. Like the mammalian and plant GRP94s, the L. infantum GRP94 sequence possesses both an N-terminal signal peptide and a putative endoplasmic reticulum retention signal, consisting of the C-terminal tetrapeptide EDDL. Thus, L. infantum is the first protozoan organism in which GRP94 has been identified. Southern blot analysis has indicated that this protein is encoded by a single-copy gene. The L. infantum GRP94 gene was expressed in Escherichia coli and the recombinant protein used to evaluate its antigenicity and immunogenicity. Eighty-four percent of sera from dogs with visceral leishmaniasis reacted with the protein, indicating that GRP94 is a potent immunogen during Leishmania infection. Given the immunogenic and antigenic properties shown by the L. infantum GRP94, we think that this protein constitutes a valuable molecule for diagnostic purposes and a potential candidate for studies of protective immunogenicity. PMID- 11052870 TI - Entamoeba histolytica: diminution of erythrophagocytosis, phospholipase A(2), and hemolytic activities is related to virulence impairment in long-term axenic cultures. PMID- 11052871 TI - Saw tooth patello--femoral arthritis. AB - Four patients with an unusual form of patello-femoral arthritis are described. The characteristic feature of the condition is an erosive 'saw tooth' pattern characteristically seen on both sides of the joint. Radiologically, this pattern is best seen on the skyline view. The other principal features are a lamellar like pattern on slightly oblique lateral views, a smooth supra patellar erosion of the femur seen on the lateral view and cortical ridging of the lateral femoral condyle seen 'en face' on the AP projection. A possible mechanism to account for the radiological features is proposed. Anbarasu, A., Loughran, C. F. (2000). Clinical Radiology55, 767-769. PMID- 11052872 TI - The clinical utility of spiral CT cholangiography. AB - Spiral CT cholangiography has received little attention, yet in a single breath hold spiral and with limited manipulation at the workstation it can yield high resolution images of the biliary tract. In addition it can clearly demonstrate periampullary detail and contribute some dynamic information regarding biliary excretion. The clinical utility of this technique is illustrated and discussed. Breen, D. J., Nicholson, A. A. (2000). Clinical Radiology55, 733-739. PMID- 11052873 TI - Radiology of the retropharyngeal space. AB - The retropharyngeal space extends from the skull base to the T4 vertebral level and contains the retropharyngeal nodes superiorly and fatty tissue elsewhere. This space is important as it is a potential route for the spread of infection and malignancy. This pictorial essay outlines the anatomy of the retropharyngeal space and illustrates the various disease processes that may be seen in region.Chong, V. F. H., Fan, Y. F. (2000). Clinical Radiology55, 740-748. PMID- 11052874 TI - Treatment of iatrogenic femoral artery pseudoaneurysms using ultrasound-guided injection of thrombin. AB - AIM: To evaluate the use of ultrasound-guided percutaneous injection of thrombin for treatment of femoral artery pseudoaneurysms. METHOD: Nine patients with a confirmed femoral false aneurysm were included in the study. 0.5-1 ml of a 2000 U/ml solution of activated bovine thrombin was injected under ultrasound visualization into the neck of the aneurysm to induce thrombosis. The parent artery and adjacent major vessels were checked during and after the procedure to exclude propagation of thrombus. A check ultrasound examination was undertaken on the following day. RESULTS: Eight patients were successfully treated by a single injection. One patient required a second injection due to recurrence of their pseudoaneurysm 4 days after the initial treatment. The procedure was well tolerated in all cases and no complications were encountered. CONCLUSION: This small series provides further evidence that ultrasound-guided thrombin injection is a promising new method for the treatment of femoral false aneurysms.Hughes, M. J. et al. (2000). Clinical Radiology55, 749-751. PMID- 11052875 TI - MRI changes in myocarditis--evaluation with spin echo, cine MR angiography and contrast enhanced spin echo imaging. AB - AIM: Myocarditis is probably under-diagnosed with clinical criteria generally used for diagnosis. Magnetic resonance imaging (MRI) has shown promise in detecting heart muscle disorders and we set out to assess the role of cine magnetic resonance angiography (MRA) and contrast enhancement in myocarditis, as there is a need for a non-invasive tool that can aid prognosis and follow-up. MATERIALS AND METHODS: Twenty patients were evaluated with T1 SE pre- and post gadolinium enhancement and cine MRA. Four patients were histologically proven to have myocarditis, eight others were diagnosed as having myocarditis by clinical criteria and eight did not have myocarditis. Images were evaluated in a blinded fashion for regional wall motion abnormality and contrast enhancement pattern. Analysis of contrast enhancement by signal intensity measurement was also performed. RESULTS: Focal myocardial enhancement with associated regional wall motion abnormality correlated with myocarditis in 10 out of 12 patients, two patients with abnormal focal enhancement alone also clinically had myocarditis. None of the non-myocarditis patients showed abnormal focal enhancement. Enhancement analysis suggests that focal corrected myocardial enhancement of > 40% is abnormal. CONCLUSION: In the correct clinical context, focal myocardial enhancement on spin echo MRI strongly supports a diagnosis of myocarditis, especially when associated with regional wall motion abnormality.Roditi, G. H. (2000). Clinical Radiology55, 752-758. PMID- 11052876 TI - The role of colour flow Doppler in the investigation of the salivary gland tumour. AB - AIMS: Ultrasound is a highly effective imaging technique to determine salivary gland tumours and may help to identify many benign lesions. The aim of this study is to evaluate whether colour Doppler is able to further differentiate the malignant tumour. METHODS: Fifty-six patients with salivary gland lesions were prospectively assessed using ultrasound imaging with colour flow and power Doppler. The peak systolic velocity (PSV) was measured and the pulsatility index (PI) and resistive index (RI) calculations were performed on the pulsed wave traces. The real time ultrasound morphology and the Doppler information were correlated with the histology. RESULTS: In 18 of the 56 patients, no internal colour flow or power Doppler changes could be detected. The real time ultrasound morphology diagnosed benign disease with sensitivity of 89.7% with specificity of 57.1%. The positive predictive value was 93.6%. There were no significant differences in the colour Doppler appearances in terms of vessel type or intratumour distribution which could separate benign from malignant conditions. However, there was statistical discrimination for PI and RI values (P = 0.0006, P = 0.0002, respectively). No malignant lesions were seen when the PI was less than 1.8 and RI was less than 0.8. The PSV was elevated in several cases (> 50 cm per s) but there was no statistical correlation with malignancy. CONCLUSION: The risk of malignancy increases by a third when the colour Doppler demonstrates increased intratumour vascular resistance (RI > 0.8 and PI > 1.8), with positive predictive value of 97.3% (sensitivity 75.5%, specificity 85.7%).Bradley, M. J. (2000). Clinical Radiology55, 759-762. PMID- 11052877 TI - Multiple 14G stereotactic core biopsies in the diagnosis of mammographically detected stellate lesions of the breast. AB - AIM: The aim of this retrospective study was to measure the accuracy of stereotactic guided 14 gauge core biopsy in distinguishing between benign and malignant causes of a mammographically detected stellate breast lesion and to assess the impact of the number of core samples taken on the sensitivity for detection of malignancy. MATERIALS AND METHODS: Seventy-two patients with mammographically detected stellate lesions of the breast formed the study group. All patients in the study group underwent multiple 14 gauge core biopsies using prone stereotactic breast biopsy equipment. The diagnostic accuracy of the technique was measured by retrospectively comparing the outcome with the core biopsy results. The result of each core sample was separately recorded to allow analysis of the effect of increasing the number of samples on accuracy. RESULTS: Nine of 72 (12%) did not have surgery. Forty of 72 (56%) had a benign surgical outcome and 23/72 (32%) a malignant surgical outcome [7/72 (10%) non-invasive, 16/72 (22%) invasive carcinoma]. The absolute sensitivity for multiple stereotactic guided core biopsies of stellate lesions for the detection of malignancy was 78% with a complete sensitivity of 100%. The sensitivity for the detection of invasive carcinoma was 94% (15 out of 16 patients). No statistically significant improvement in sensitivity was shown for multiple samples vs one sample, but in two patients, malignant tissue was only found in core samples 6-9, the first five cores showing atypia only. CONCLUSION: Multiple stereotactic guided 14 gauge core biopsies accurately distinguish malignant from benign causes of stellate breast lesions. When core biopsy histology is malignant, therapeutic surgery can be planned. When the core biopsy shows typical features of a benign radial scar, diagnostic surgical excision may not be required to confirm the diagnosis.Kirwan, S. E., (2000). Clinical Radiology55, 763-766. PMID- 11052878 TI - Sonographic evaluation of thyroglossal duct cysts in children. AB - BACKGROUND AND AIMS: Thyroglossal duct cysts (TDC) in children have a variable sonographic appearance. Some reports have suggested that TDCs appear on ultrasound as well defined, cystic masses with thin walls and posterior enhancement, whereas others have documented a heterogeneous echopattern within these lesions. In our experience, although TDCs in children have a variable ultrasound appearance, the most common appearance is that of a pseudosolid mass closely related to the hyoid bone. In this study we report on 23 patients with thyroglossal duct cysts and document the ultrasonic patterns. PATIENTS AND METHODS: All patients in whom the diagnosis of TDC was made clinically (by at least two head and neck surgeons) and in whom ultrasound detected a cystic mass related to the hyoid bone, were included in this study. Sonograms of 23 children with TDCs were reviewed. The features evaluated included their location, internal echogenicity, posterior enhancement, the presence of septa, a solid component and a fistulous tract. The echopattern was not correlated with the biopsy results. RESULTS: Three patterns of TDCs were identified: anechoic (13%); pseudosolid (56.5%); and a heterogeneous pattern (30.5%). The majority were midline (82.6%), showed posterior enhancement (56.5%), and had thin walls (82.6%). CONCLUSION: On ultrasound, TDCs in children are not simple cysts but have a complex pattern ranging from a typical anechoic cyst to a pseudosolid appearance (most common).Ahuja, A. T. (2000). Clinical Radiology55, 770-774. PMID- 11052879 TI - Complications after laryngeal surgery: videofluoroscopic evaluation of 120 patients. AB - AIM: Videofluoroscopic assessment of the spectrum and incidence of swallowing complications after state-of-the-art laryngeal cancer surgery. MATERIALS AND METHODS: We retrospectively studied videofluoroscopic examinations of 120 patients (94 men, 26 women; mean age, 58 years) with suspected complications after laryngeal resection (partial laryngectomy, 65; total laryngectomy, 55). Swallowing function (i.e., oral bolus control, laryngeal elevation and closure, presence of pharyngeal residue, aspiration) and structural abnormalities such as strictures, fistulas and tumour recurrence were assessed by videofluoroscopy. RESULTS: Abnormalities were found in 110 patients, including strictures in nine, fistulas in six and mass lesions in 13 patients. Aspiration was found in 63 patients overall (partial laryngectomy, 61/65; total laryngectomy, 2/55), occurring before swallowing in five, during swallowing in 34, after swallowing in nine and at more than one phase in 15 patients. Pharyngeal paresis was detected in three and pharyngeal weakness in 19 patients. Pharyngo-oesophageal sphincter dysfunction was observed in 10 cases. CONCLUSION: Aspiration is a very common complication after partial laryngeal resection. It is mainly caused by incomplete laryngeal closure, sphincter dysfunction or pharyngeal pooling. Videofluoroscopy is the only radiological technique able to identify both disordered swallowing function and structural changes after laryngeal resection. Detection of these complications is crucial for appropriate further therapy.Kreuzer, S. H. (2000). Clinical Radiology55, 775-781. PMID- 11052880 TI - A comparison of digital and screen-film mammography using quality control phantoms. AB - AIM: To compare the performance of a direct digital mammography system with normal-view and magnified-view conventional screen-film methods using quality control phantoms. MATERIALS AND METHODS: Using a Siemens Mammomat((R))3000 and an Opdima((R))digital spot imaging and biopsy attachment, film and direct digital images of two phantoms [DuPont and TOR (MAM)] were obtained under normal operating conditions. These were assessed by three groups of observers with differing expertise - radiologists, radiographers and medical physicists. Each observer was asked to compare the direct digital image with films taken in standard view and magnified view, providing scores for object visibility and confidence. For the digital images, observers were allowed to vary the image presentation parameters. RESULTS: Both phantoms showed that overall the direct digital view and the magnified view film performed significantly better (P < 0.05) than standard view film. For certain small or low contrast objects the differences became very highly significant (P < 0.001). CONCLUSION: Only the TOR (MAM) phantom showed any significant difference between digital and magnified modalities, with magnified views performing better for fine, faint filaments and digital acquisition better for low contrast objects. Almost no difference existed between the three observer groups. Undrill, P. E. (2000). Clinical Radiology53, 782-790. PMID- 11052881 TI - The effect of resecting the primary tumour on the Doppler Perfusion Index in patients with colorectal cancer. AB - AIM: In patients undergoing apparently curative resection for colorectal cancer, an elevated Doppler Perfusion Index (DPI; ratio of hepatic arterial to total liver blood flow) before surgery is associated with a high risk of recurrence. The role of the primary tumour in inducing and sustaining these blood flow changes is unknown. The aim of this study was to assess the impact of removing the primary tumour on the DPI. MATERIALS AND METHODS: Using duplex/colour Doppler sonography, the DPI was measured both before and after surgery (median, 9 months) in 14 patients undergoing apparently curative resection for colorectal cancer. RESULTS: In the five patients with a normal pre-operative DPI (< 0.30), there was no significant change following surgery. In the nine patients with an abnormal pre-operative DPI, there was a small but significant fall from 0.38 (SEM 0.02) to 0.33 (0.02) following surgery (P = 0.04). However, DPI values remained abnormally elevated in seven of these nine patients. CONCLUSION: These results suggest that the primary tumour plays a relatively minor role in inducing an abnormally elevated DPI in patients undergoing apparently curative resection for colorectal cancer.Oppo, K. (2000). Clinical Radiology55, 791-793. PMID- 11052882 TI - Subdiaphragmatic pulmonary sequestrian simulating metastatic testicular cancer. PMID- 11052883 TI - CT demonstration of an unusual cause of biliary obstruction in a patient with peripheral neurofibromatosis. PMID- 11052884 TI - Spontaneous pneumopericardium secondary to penetrating benign gastric ulcer. PMID- 11052885 TI - Carbon dioxide and gadodiamide as the contrast agents for diagnosis and embolization of a post-biopsy arteriovenous fistula in a renal allograft. PMID- 11052886 TI - Hepatic infarction due to thrombotic angiitis; MR appearance. PMID- 11052887 TI - Blunt cardiac rupture: value of contrast-enhanced spiral CT. PMID- 11052888 TI - An Atlas of Diagnostic Radiology in Gastroenterology. AB - Nolan, D. J. (2000). Clinical Radiology55, 809. Copyright 2000 The Royal College of Radiologists. PMID- 11052889 TI - Monitoring biomolecular interactions by time-lapse atomic force microscopy. AB - The atomic force microscope (AFM) is a unique imaging tool that enables the tracking of single macromolecule events in response to physiological effectors and pharmacological stimuli. Direct correlation can therefore be made between structural and functional states of individual biomolecules in an aqueous environment. This review explores how time-lapse AFM has been used to learn more about normal and disease-associated biological processes. Three specific examples have been chosen to illustrate the capabilities of this technique. In the cell, actin polymerizes into filaments, depolymerizes, and undergoes interactions with numerous effector molecules (i.e., severing, capping, depolymerizing, bundling, and cross-linking proteins) in response to many different stimuli. Such events are critical for the function and maintenance of the molecular machinery of muscle contraction and the dynamic organization of the cytoskeleton. One goal is to use time-lapse AFM to examine and manipulate some of these events in vitro, in order to learn more about how these processes occur in the cell. Aberrant protein polymerization into amyloid fibrils occurs in a multitude of diseases, including Alzheimer's and type 2 diabetes. Local amyloid deposits may cause organ dysfunction and cell death; hence, it is of interest to learn how to interfere with fibril formation. One application of time-lapse AFM in this area has been the direct visualization of amyloid fibril growth in vitro. This experimental approach holds promise for the future testing of potential therapeutic drugs, for example, by directly visualizing at which level of fibril assembly (i.e., nucleation, elongation, branching, or lateral association of protofibrils) a given active compound will interfere. Nuclear pore complexes (NPCs) are large supramolecular assemblies embedded in the nuclear envelope. Transport of ions, small molecules, proteins, RNAs, and RNP particles in and out of the nucleus occurs via NPCs. Time-lapse AFM has been used to structurally visualize the response of individual NPC particles to various chemical and physical effectors known to interfere with nucleocytoplasmic transport. Taken together, such time lapse AFM studies could provide novel insights into the molecular mechanisms of fundamental biological processes under both normal and pathological conditions at the single molecule level. PMID- 11052890 TI - Crystallisation of CP43, a chlorophyll binding protein of photosystem II: an electron microscopy analysis of molecular packing. AB - Microcrystals of the chlorophyll binding protein, CP43, isolated from spinach thylakoid membranes have been studied by electron microscopy both in negative stain and in vitreous ice. Image analyses of three characteristic views show that the crystals are built of five different layers perpendicular to the c-axis. Each layer consists of different orientations of the CP43 protein. The unit cell derived from the end-on view (looking down the c-axis) shows an angle of 120 degrees, suggesting a threefold rotational symmetry. Both negative staining and cryo data are consistent with a hexagonal crystal lattice. Interpretation of the arrangement of the CP43 protein within this crystal lattice can be made based on 8- and 9-A electron crystallographic structures previously published that provide a model for the organisation of the transmembrane helices of CP43. Overall the analysis presented is consistent with X-ray diffraction data obtained from larger CP43 crystals and forms a framework on which to base further structural studies of this chlorophyll binding protein. PMID- 11052891 TI - Interaction of myosin LYS-553 with the C-terminus and DNase I-binding loop of actin examined by fluorescence resonance energy transfer. AB - Fluorescence resonance energy transfer (FRET) experiments were carried out in the absence of nucleotide (rigor) or in the presence of MgADP between fluorescent donor probes (IAEDANS (5((((2-iodoacetyl)amino)ethyl)amino)-naphthalene-1 sulfonic acid) at Cys-374 or DANSYL (5-dimethylamino naphthalene-1-(N-(5 aminopentyl))sulfonamide) at Gln-41 of actin and acceptor molecules (FHS (6 [fluorescein-5(and 6)-carboxamido] hexanoic acid succinimidyl ester) at Lys-553 of skeletal muscle myosin subfragment 1. The critical Forster distance (R(0)) was determined to be 44 and 38 A for the IAEDANS-FHS and DANSYL-FHS donor-acceptor pairs, respectively. The efficiency of energy transfer between the acceptor molecules at Lys-553 of myosin and donor probes at Cys-374 or Gln-41 of actin was calculated to be 0.78 +/- 0.01 or 0.94 +/- 0.01, respectively, corresponding to distances of 35.6 +/- 0.4 A and 24.0 +/- 1.6 A, respectively. MgADP had no significant effect on the distances observed in rigor. Thus, rearrangements in the acto-myosin interface are likely to occur elsewhere than in the lower 50-kDa subdomain of myosin as its affinity for actin is weakened by MgADP binding. PMID- 11052892 TI - Polymerization of deoxy-sickle cell hemoglobin in high-phosphate buffer. AB - Deoxy-sicklecell hemoglobin (HbS) polymerizes in 0.05 M phosphate buffer to form long helical fibers. The reaction typically occurs when the concentration of HbS is about 165 mg/ml. Polymerization produces a variety of polymorphic forms. The structure of the fibers can be probed by using site-directed mutants to examine the effect of altering the residues involved in intermolecular interactions. Polymerization can also be induced in the presence of 1.5 M phosphate buffer. Under these conditions polymerization occurs at much lower concentrations (ca. 5 mg/ml), which is advantageous when site-directed mutants are being used because only small quantities of the mutants are available. We have characterized the structure of HbS polymers formed in 1.5 M phosphate to determine how their structures are related to the polymers formed under more physiological conditions. Under both sets of conditions fibers are the first species to form. At pHs between 6.7 and 7.3 fibers initially form bundles and then crystals. At lower pHs fibers form macrofibers and then crystals. Fourier transforms of micrographs of the polymers formed in 1.5 M phosphate display the 32- and 64-A( 1) periodicity characteristic of fibers formed in 0.05 M phosphate buffer. The 64 A(-1) layer line is less prominent in Fourier transforms of negatively stained fibers formed in 1.5 M phosphate possibly because salt interferes with staining of the fibers. However, micrographs and Fourier transforms of frozen hydrated fibers formed in high and low phosphate display the same periodicities. Under both sets of reaction conditions HbS polymers form crystals with the same unit cell parameters as Wishner-Love crystals (a = 64 A, b = 185 A, c = 53 A). Some of the polymerization intermediates were examined in the frozen-hydrated state in order to determine whether their structures were significantly perturbed by negative staining. We have also carried out reconstructions of the frozen hydrated fibers in high and low phosphate to compare their molecular coordinates. The helical projection of the reconstructions in low phosphate shows the expected 14-strand structure. In high phosphate the 14-strand fibers are also formed and their molecular coordinates are the same (within experimental error) as those of fibers formed in 0.05 M phosphate. In addition, the reconstructions of high phosphate fibers reveal a new minor variant of fiber containing 10 strands. The polymerization products in 1.5 M phosphate buffer were generally indistinguishable from those formed in 0.05 M phosphate buffer. Micrographs of frozen hydrated specimens have facilitated the interpretation of previously published micrographs using negative staining. PMID- 11052893 TI - The structure of ferritin cores determined by electron nanodiffraction. AB - Electron nanodiffraction, with a 100-keV electron beam less than 1 nm in diameter, has been used to obtain single-crystal diffraction patterns from individual iron-containing cores of ferritin molecules. We show that, while a majority of the cores have a hexagonal structure somewhat similar to the major phase in the mineral ferrihydrite, as previously assumed, several minor phases are present including some that are similar in structure to the iron oxides magnetite and hematite and also some composed of highly disordered material. In general, each core consists of one single crystal of one phase. PMID- 11052894 TI - MARCKS-related protein binds to actin without significantly affecting actin polymerization or network structure. Myristoylated alanine-rich C kinase substrate. AB - Actinis a 42-kDa protein which, due to its ability to polymerize into filaments (F-actin), is one of the major constituents of the cytoskeleton. It has been proposed that MARCKS (an acronym for myristoylated alanine-rich C kinase substrate) proteins play an important role in regulating the structure and mechanical properties of the actin cytoskeleton by cross-linking actin filaments. We have recently reported that peptides corresponding to the effector domain of MARCKS proteins promote actin polymerization and cause massive bundling of actin filaments. We now investigate the effect of MARCKS-related protein, a 20-kDa member of the MARCKS family, on both filament structure and the kinetics of actin polymerization in vitro. Our experiments document that MRP binds to F-actin with micromolar affinity and that the myristoyl chain at the N-terminus of MRP is not required for this interaction. In marked contrast to the effector peptide, binding of MRP is not accompanied by an acceleration of actin polymerization kinetics, and we also could not reliably observe an actin cross-linking activity of MRP. PMID- 11052895 TI - Truncation of vertebrate striated muscle myosin light chains disturbs calcium induced structural transitions in synthetic myosin filaments. AB - Electron microscopy and negative staining techniques have been used to show that the proteolytic removal of 13 amino acids from the N-terminus of essential light chain 1 and 19 amino acids from the N-terminus of the regulatory light chain of rabbit skeletal and cardiac muscle myosins destroys Ca(2+)-induced reversible movement of subfragment-2 (S2) with heads (S1) away from the backbone of synthetic myosin filaments observed for control assemblies of the myosin under near physiological conditions. This is the direct demonstration of the contribution of the S2 movement to the Ca(2+)-sensitive structural behavior of rabbit cardiac and skeletal myosin filaments and of the necessity of intact light chains for this movement. In muscle, such a mobility might play an important role in proper functioning of the myosin filaments. The impairment of the Ca(2+) dependent structural behavior of S2 with S1 on the surface of the synthetic myosin filaments observed by us may be of direct relevance to some cardiomyopathies, which are accompanied by proteolytic breakdown or dissociation of myosin light chains. PMID- 11052896 TI - Surface topography of the p3 and p6 annexin V crystal forms determined by atomic force microscopy. AB - Annexin V is a member of a family of structurally homologous proteins sharing the ability to bind to negatively charged phospholipid membranes in a Ca(2+) dependent manner. The structure of the soluble form of annexin V has been solved by X-ray crystallography, while electron crystallography of two-dimensional (2D) crystals has been used to reveal the structure of its membrane-bound form. Two 2D crystal forms of annexin V have been reported to date, with either p6 or p3 symmetry. Atomic force microscopy has previously been used to investigate the growth and the topography of the p6 crystal form on supported phospholipid bilayers (Reviakine et al., 1998). The surface structure of the second crystal form, p3, is presented in this study, along with an improved topographic map of the p6 crystal form. The observed topography is correlated with the structure determined by X-ray crystallography. PMID- 11052897 TI - Crystallization and initial X-ray diffraction characterization of complexes of FxFG nucleoporin repeats with nuclear transport factors. AB - NTF2 and importin-beta are transport factors that mediate nuclear protein import and which interact with nuclear pore proteins (nucleoporins) during translocation from the cytoplasm to the nucleus through nuclear pore complexes. We employed a native gel electrophoresis method to assess the interaction of nucleoporin constructs that contain FxFG sequence repeats with NTF2 and truncation mutants of importin-beta to determine suitable fragments for crystallization. Based on these data, we obtained crystals of complexes between yeast NTF2 and a construct containing five FxFG nucleoporin repeats from the yeast nucleoporin Nsp1p and between a construct containing residues 1-442 of human importin-beta and the same nucleoporin construct. The yeast NTF2-nucleoporin crystals have trigonal symmetry and diffract past 2.8 A resolution using synchrotron radiation, whereas the importin-beta-nucleoporin complex crystals have P2(1)2(1)2 orthorhombic symmetry and diffract past 3.2 A resolution. PMID- 11052898 TI - Fads and fashion. PMID- 11052899 TI - The role of physiotherapy in the prevention and treatment of osteoporosis. AB - Osteoporosis is an increasing public health problem that causes loss of life and reduced quality of life in sufferers. Strategies to improve bone density and reduce the likelihood of falls are important in the prevention of osteoporosis. Physiotherapists have a role to play in this condition through exercise prescription, therapeutic modalities, specific techniques and education. Appropriate treatment goals can be established following a thorough assessment of signs and symptoms, risk factors for osteoporosis and functional status. Levels of bone density measured from dual energy X-ray absorptiometry can help guide patient management. Since the aim is to maximize peak bone mass in children and adolescents, participation in a variety of high-impact activities should be encouraged. In the middle adult years, small increases in bone mass may be achieved by structured weight-training and weight-bearing exercise. In the older adult years, particularly if osteopenia or osteoporosis is present, the aim is to conserve bone mass, reduce the risk of falls, promote extended posture, reduce pain, and improve mobility and function. PMID- 11052900 TI - An investigation to compare the effectiveness of carpal bone mobilisation and neurodynamic mobilisation as methods of treatment for carpal tunnel syndrome. AB - Carpal tunnel syndrome is the most common peripheral entrapment neuropathy. There is little literature available that addresses the management of this condition, which may partly explain why physiotherapy is often overlooked as a treatment approach in its management. This study investigated the effects of two manual therapy techniques in the treatment of patients experiencing carpal tunnel syndrome. An experimental different subject design compared three groups of subjects in three different conditions (two treatment interventions and one control group). Each group consisted of seven patients. The objectives of the study were: (1) to investigate differences between treated and untreated groups; (2) to investigate differences in the effectiveness of treatment I (median nerve mobilization) compared with treatment II (carpal bone mobilization). Measurements were taken applying several measurement tools, including active range of wrist movement (ROM flexion and extension), upper limb tension test with a median nerve bias (ULTT2a), three different scales to evaluate pain perception and function, and lastly numbers of patients continuing to surgery in each group were compared. In visual terms a clear trend was demonstrated between subjects who received treatment compared to those who were not treated, in particular the descriptive analysis of results for ULTT2a and numbers of patients continuing to surgery. When analysed statistically, less could be concluded. Only scores on a Pain Relief Scale (P<0.01) demonstrated highly significant differences between the three groups when analyzed using Kruskal-Wallis Test. In exploring the results of the two intervention groups, no statistically significant difference in effectiveness of treatment was demonstrated between carpal bone mobilization and median nerve mobilization. PMID- 11052901 TI - PEDro. A database of randomized trials and systematic reviews in physiotherapy. AB - This paper describes PEDro, the Physiotherapy Evidence Database. PEDro is a web based database of randomized controlled trials and systematic reviews in physiotherapy. It can be accessed free of charge at http://ptwww.cchs.usyd.edu.au/pedro. The database contains bibliographic details and abstracts of most English-language randomized trials and systematic reviews in physiotherapy, and of many trials and reviews in other languages. Trials on the database are rated on the basis of their methodological quality so that users of the database can quickly identify trials of high quality. Trials and systematic reviews are extensively indexed to facilitate searching. PEDro provides an important information resource to support evidence-based clinical practice. PMID- 11052902 TI - Bilateral buttock pain caused by aortic stenosis: a case report of claudication of the buttock. PMID- 11052904 TI - Diary of events PMID- 11052903 TI - A newly developed spinal simulator. AB - A number of studies indicate poor intra-therapist and inter-therapist reliability in the performance of graded, passive oscillatory movements to the lumbar spine. However, it has been suggested that therapists can be trained to be more consistent in their performance of these techniques if given reliable quantitative feedback. The intention of this study was to develop equipment, analogous to the lumbar spine that could be used for both teaching and research purposes. Equipment has been updated and connected to a personal IBM compatible computer. Custom designed software allows concurrent and accurate feedback to students on their performance and in a form suitable for advanced data analysis using statistical packages. The uses and implications of this equipment are discussed. PMID- 11052905 TI - Evidence based practice-review and clinical trials. PMID- 11052906 TI - Pulmonary surfactant in innate immunity and the pathogenesis of tuberculosis. AB - Components of the innate immune system serve to protect the host from invading pathogens prior to the generation of a directed immune response, and influence the manner in which the directed immune response develops. The pulmonary surfactant system consists of a complex array of proteins and lipids that reduce surface tension of the alveoli, and appears to play an essential role in innate immunity. Investigators have recently gained insight into the interactions between components of the surfactant system and the respiratory pathogen Mycobacterium tuberculosis. It is likely that pulmonary surfactant and other innate immune determinants play significant roles in the pathogenesis of tuberculosis. PMID- 11052907 TI - An esat6 knockout mutant of Mycobacterium bovis produced by homologous recombination will contribute to the development of a live tuberculosis vaccine. AB - SETTING: Strains of the Mycobacterium tuberculosis complex are being rationally attenuated in order to develop better tuberculosis vaccines than BCG, and it would be helpful if new vaccines lacked an immunogenic protein which could be used as a skin test reagent for determining infection status. OBJECTIVE: To delete the esat6 gene from a virulent Mycobacterium bovis strain and determine (i) whether this mutant sensitizes guinea pigs to a skin test based on ESAT6 and (ii) what effect this has on the virulence of M. bovis. DESIGN: An homologous recombination technique was used to produce an esat6 knockout mutant of a virulent strain of M. bovis. Guinea pigs were inoculated with either the mutant or parent strain and their reactivity in intradermal skin tests was determined to bovine purified protein derivative (PPD) and recombinant ESAT6 protein. RESULTS: Production of an esat6 knockout strain was demonstrated by Southern blot hybridization and the polymerase chain reaction. Guinea pigs inoculated with either the esat6 knockout strain or its virulent parent had positive skin test reactions to PPD but only animal inoculated with the parent strain had positive skin test reactions to ESAT6. Gross pathology, histopathology and mycobacterial culture of tissues indicated that the knockout strain was less virulent than its parent. CONCLUSION: If an effective live tuberculosis vaccine can be produced by inactivation of virulence genes in M. bovis, then prior or subsequent knockout of the esat6 gene could contribute to the loss of virulence and enable the development of a test to distinguish between vaccinated and infected animals. PMID- 11052908 TI - TB PCR in the early diagnosis of tuberculous meningitis: evaluation of the Roche semi-automated COBAS Amplicor MTB test with reference to the manual Amplicor MTB PCR test. . AB - SETTING: Cecilia Makiwane Hospital, Mdantsane, Eastern Cape, Republic of South Africa. OBJECTIVE: To assess the role of the semi-automated Roche COBAS AMPLICOR(TM)Mycobacterium tuberculosis PCR test in the diagnosis of tuberculous meningitis (TBM). DESIGN: Eighty-three specimens of cerebrospinal fluid (CSF) were collected prospectively from 69 patients with suspected TBM. The COBAS AMPLICOR TB PCR test was compared with the manual AMPLICOR(TM)TB PCR test, clinical and cerebrospinal fluid (CSF) findings, direct ZN smear and radiometric TB culture. RESULTS: CSF from 7/40 (17.5%) patients treated for TBM were positive by TB COBAS AMPLICOR(TM). The sensitivity of the test was not significantly different (p=0.375) from the manual TB AMPLICOR(TM)PCR test. The comparative sensitivities of the TB COBAS AMPLICOR(TM)PCR and the manual AMPLICOR PCR for detecting cases of definite and probable TBM from CSF collected within 9 days of commencing antituberculosis treatment were 40% and 60% respectively. All 29 patients not treated for TBM were negative by COBAS AMPLICOR(TM), giving a specificity of 100%. CONCLUSION: The COBAS AMPLICOR(TM)TB PCR test is a rapid and highly specific diagnostic test for TBM. However, there was a non-significant trend favouring slightly greater sensitivity using the manual AMPLICOR(TM)TB PCR test. PMID- 11052909 TI - Monocyte-derived macrophage cytokine responses induced by M. bovis BCG. AB - SETTING: One important aspect of macrophage function is the production of inflammatory and anti-inflammatory cytokines, which in turn affect the survival of intracellular organisms such as mycobacteria. OBJECTIVE: To determine the relationship between phagocytosis of mycobacteria and expression of intracellular cytokines. DESIGN: Phagocytosis and cytokine production were studied simultaneously within human monocyte-derived macrophages (MDMs) from healthy donors using fluorescent labelling of M. bovis BCG and flow cytometry. RESULTS: At a range of infection ratios (5:1, 1:1, 0.2:1) TNF- alpha, IL-10, IL-6 and IL 12 were all produced in a dose-dependent manner. At an infection ratio representative of the in vivo situation (1:1), cytokine production was induced in both MDMs containing intracellular M. bovis BCG and in uninfected bystander MDMs. Phagocytosis increased over time, but there was considerable donor variation: the proportions of cells containing one or more mycobacterium were 15.4+/-14.8% (mean+/-SD) at 4 h and 32.7+/-21.1% at 24 h (n=19). Analysis of cytokine production by MDMs not containing mycobacteria (bystander cells) at 4 h revealed that these uninfected cells produced 79+/-6.6% of the TNF- alpha, 53.9+/-40.0% of the IL-10 and 64.2+/-12.4% of the IL-12. By 20 h these proportions had decreased to 57+/-13.5%, 30.9+/-7.4% and 45.5+/-13.3% respectively. CONCLUSION: Both infected and bystander MDMs can be stimulated to produce cytokines in response to M. bovis BCG, indicating that the ability of MDMs to produce cytokines is not necessarily dependent on the ability to phagocytose mycobacteria. PMID- 11052910 TI - Efficacy of Amplicor PCR for the diagnosis of tuberculosis in respiratory specimens other than sputum. AB - A total of 832 respiratory specimens not including the sputum (402 bronchial lavages, 241 bronchial brushing specimens, 136 pumping lavages, 41 pleural effusions, and 12 others) from 462 patients were assayed using the Roche Amplicor Mycobacterium tuberculosis test for amplification and identification of M. tuberculosis, M. avium and M. intracellulare (Amplicor PCR). The results were compared with those obtained using conventional microscopy and cultivation methods. Each patient had little or no sputum and showed an abnormal chest X-ray shadowing of unknown cause. No patients had previously undergone antituberculous therapy. Of the specimens obtained, 24 were both PCR and culture positive, 786 were both PCR and culture negative, 11 were PCR positive and culture negative, and 11 were PCR negative and culture positive. Based on these results, the sensitivity and specificity of Amplicor PCR were determined to be 68.67% and 98.6%, respectively, when compared with culture of respiratory specimens not including the sputum. After correcting for discrepancies due to differences in patient clinical data, the sensitivity of Amplicor PCR was found to be 68.6%, and the specificity to be 99.9%; the corresponding values for culture were 66.7% and 100%, and those for smear were 9.8% and 100%. Thus, Amplicor PCR was shown to possess a similar sensitivity to culture and to be a highly specific technique for the diagnosis of tuberculosis in the respiratory system using non-sputum specimens within hours in patients showing little or no sputum and abnormal chest X-ray shadowing of an indeterminant cause. PMID- 11052911 TI - Influence of relative humidity on particle size and UV sensitivity of Serratia marcescens and Mycobacterium bovis BCG aerosols. AB - SETTING: A study of Serratia marcescens and BCG aerosols. OBJECTIVE: To evaluate the effect of relative humidity (RH) on (1) the particle size and (2) sensitivity of 254nm germicidal ultraviolet (UV) irradiation. METHODS: We built a RH controlled experimental chamber into which bacteria were aerosolized, exposed to varying amounts of UV irradiance over measured time periods, and quantitatively evaluated for viability. Aerosolized Serratia marcescens and bacille Calmette Guerin (BCG) were subject to UV doses ranging from 57-829 microW. sec/cm(2), and sampled with a six-stage Andersen culture plate impactor at RHs ranging from 25 95%. RESULTS: Percent survival for both organisms was inversely related to UV dose. Serratia marcescens was more susceptible to UV than BCG under all conditions. More than 95% of the bacterial aerosol particles were 1.1-4.7 microm in aerodynamic diameter, and particles sizes increased from low (25-36%) to high (85-95%) RH. The count median diameter ranged from 1.9-2.6 microm for Serratia marcescens and from 2.2-2.7 microm for BCG as RH increased. For both Serratia marcescens and BCG, resistance to UV increased as RH increased. The UV resistance of both Serratia marcescens and BCG aerosols dramatically increased at RH higher than 85%. CONCLUSIONS: Our results indicate that differences in UV dose, kinds of microorganisms, airborne particle size and RH affect UV susceptibility. PMID- 11052912 TI - The development of wildlife control strategies for eradication of tuberculosis in cattle in Ireland. AB - Wildlife species, such as badgers, act as maintenance hosts for Mycobacterium bovis and contribute to the spread and persistence of tuberculosis in associated cattle populations. In areas in which there is a tuberculosis problem affecting a number of herds, the involvement of infected wildlife in the introduction of M. bovis infection into herds act as a constraint to eradication of the disease. Epidemiological evidence demonstrates a high prevalence of tuberculosis in badgers, and controlled studies involving comprehensive badger removal have shown that this strategy can serve to significantly reduce cattle reactor rates in the targeted areas. However, as the badger is a protected wildlife species, alternative strategies are required to combat the disease. Targeted vaccination of wildlife species against tuberculosis is an option which, if successfully employed, could directly facilitate the advancement of bovine tuberculosis eradication in affected areas. Any proposed vaccination programme would need to be undertaken against the background of an exhaustive investigation of the cattle and herd management-related factors, and take account of environmental issues. PMID- 11052913 TI - Deletion of the putative antioxidant noxR1 does not alter the virulence of Mycobacterium tuberculosis H37Rv. AB - SETTING: The cloned M. tuberculosis noxR1 gene has been shown to confer resistance to reactive nitrogen intermediates (RNI) and reactive oxygen intermediates (ROI) upon Escherichia coli and Mycobacterium smegmatis. OBJECTIVE: To investigate the role of noxR1 in resistance to RNI and virulence of M. tuberculosis. DESIGN: The noxR1 gene was deleted from M. bovis BCG and M. tuberculosis H37Rv by allelic exchange. The mutants were compared to wild type strains with respect to resistance to chemically generated RNI. The virulence of the M. tuberculosis mutant was investigated in a murine model of infection. RESULTS: The NoxR1 mutants grew normally in Sautons and 7H9 broths. The BCG mutant demonstrated decreased resistance to in vitro generated RNI compared to the wild type. Resistance to RNI could be restored to the mutant by reintroduction of the noxR1 locus on a replicating plasmid. However, deletion of noxR1 from M. tuberculosis H37Rv did not result in decreased resistance to RNI nor a difference in growth and survival of the bacterium during murine infection. CONCLUSION: The noxR1 gene locus in M. bovis BCG bestows ability to resist RNI generated in vitro. In M. tuberculosis H37Rv, however, noxR1 is either not involved in RNI resistance and virulence or is better compensated for by other mechanisms. PMID- 11052914 TI - Neurochemical quantification of human organ-specific sympathetic nervous system activity. PMID- 11052915 TI - Effects of insulin per se on neuroendocrine and metabolic counter-regulatory responses to hypoglycaemia. AB - We examined and compared findings from studies aimed at detecting and quantifying an effect of insulin per se on counter-regulatory responses to hypoglycaemia. The experimental protocols used in many of these studies were very different with regard to study design and patient population, resulting at times in inconsistencies and discrepancies. Taken together, the results from this extensive body of work clearly indicate that, at similar levels of hypoglycaemia, greater hyperinsulinaemia results in enhanced counter-regulatory responses. This enhancement includes higher circulating levels of counter-regulatory hormones (adrenaline, noradrenaline, cortisol and growth hormone, but not glucagon), more intense activation of hypoglycaemic symptoms (both neural-sympathetic and adrenal sympathetic), and greater deterioration of neuropsychological skills. The insulin induced enhancement of counter-regulatory responses is not influenced by gender, is present in several animal species, and applies to healthy subjects as well as to patients with Type I diabetes. The underlying mechanisms remain speculative, and possibly include a direct neuromodulatory effect and/or suppression of glucose utilization in various areas of the brain, which either independently or in a hierarchical fashion trigger the sequence of downstream counter-regulatory events. PMID- 11052916 TI - Comparison of two indices for forearm noradrenaline release in humans. AB - Although there is as yet no method which measures directly the neuronal release of noradrenaline in humans in vivo, the isotope dilution technique with [(3)H]noradrenaline has been applied to estimate forearm neuronal noradrenaline release into plasma. Two different equations have been developed for this purpose: one to estimate the spillover of noradrenaline into the venous effluent, and a modified formula (often referred to as the appearance rate) which may reflect more closely changes in the neuronal release of noradrenaline into the synaptic cleft, particularly during interventions that alter forearm blood flow. The present study was performed to compare the effects of two interventions known to exert contrasting actions on neuronal forearm noradrenaline release and forearm blood flow. Intra-arterial infusion of sodium nitroprusside at doses without systemic effect increases forearm blood flow, but not neuronal noradrenaline release. In contrast, lower-body negative pressure at -25 mm Hg causes forearm vasoconstriction by stimulating neuronal noradrenaline release. During sodium nitroprusside infusion, forearm noradrenaline spillover increased from 1.1+/-0.3 to 2.2+/-1.0 pmol x min(-1) x 100 ml(-1) (P<0.05), whereas the forearm noradrenaline appearance rate was unchanged. Lower-body negative pressure did not affect the forearm noradrenaline spillover rate, but increased the forearm noradrenaline appearance rate from 3.4+/-0.4 pmol x min(-1) x 100 ml(-1) at baseline to 5.0+/-0.9 pmol x min(-1) x 100 ml(-1) (P<0.05). These results indicate that the noradrenaline appearance rate provides the better approximation of changes in forearm neuronal noradrenaline release in response to stimuli which alter local blood flow. PMID- 11052917 TI - Baroreflex sensitivity in the elderly: influence of age, breathing and spectral methods. AB - Baroreflex sensitivity (BRS) has been proposed as a diagnostic parameter for neurological disorders and as a survival-prognosis parameter in diabetic and cardiac patients. Therefore reference values and the reproducibility of BRS were assessed, taking into account the possible influence of age, gender, test conditions and some analysis variants. Healthy subjects (n=191) were randomly selected from the 50-75-year-old general population (the Hoorn Study). Variations in blood pressure and heart rate were recorded non-invasively during three breathing modes: spontaneous (3 min), slow metronome (1 min; 6 breaths/min=0.1 Hz) and fast metronome (1 min; 15 breaths/min=0.25 Hz), all in a supine position. From these recordings, BRS was assessed as the transfer gain between changes in blood pressure and heart period, and as the alpha coefficient. BRS values ranged from 5.0 to 8.9 ms.mmHg(-1). Slow metronome breathing resulted in higher BRS values than fast breathing, while during spontaneous breathing BRS in the low frequency band was lower than that in the high-frequency band (respiratory origin). BRS values decreased with lower coherence criteria. BRS-alpha was significantly higher than BRS-gain. While regression analysis showed no gender differences, BRS decreased with age. Therefore age-specific reference values were calculated. The reproducibility of BRS values was in general moderate, with reliability coefficients ranging from 43 to 81% and coefficients of variation ranging from 34 to 59%. In conclusion, this study shows age, breathing mode, frequency and coherence threshold to affect measures of BRS. Therefore these factors should be considered in clinical studies; appropriate reference values are given. PMID- 11052918 TI - Vasodilator prostanoids, but not nitric oxide, may account for skeletal muscle hyperaemia in Type I diabetes mellitus. AB - We and others have previously documented increased resting and exercise-induced skeletal muscle blood flow in young subjects with Type I (insulin-dependent) diabetes mellitus compared with healthy controls. Both NO and prostanoids are important regulators of vascular tone and may therefore contribute to this hyperaemia. The aim of the present study was to determine the contribution of NO and vasodilator prostanoids to this skeletal muscle hyperaemia in diabetes. We assessed the effects of infusion into the intrabrachial artery of the cyclo oxygenase inhibitor acetylsalicylic acid (ASA; aspirin) and of the L-arginine analogue N(G)-monomethyl-L-arginine (L-NMMA) on skeLetal muscle blood flow in subjects with Type I diabetes mellitus (DM subjects) and control subjects. Blood flow was measured by venous occlusion plethysmography. Isotonic forearm exercise involved 2 min of wrist flexion and extension. Resting flow (forearm blood flow; FBF) was augmented in DM subjects, as was peak exercise-related blood flow (PFBF) and the volume repaid to the forearm 5 min after exercise (AUC 5, where AUC is area under the flow-time curve) (P<0.05), even when accounting for differences in basal flow. Infusion of L-NMMA reduced resting flow by 48% in controls (P<0.005) and by 12% in DM subjects (not significant). L-NMMA reduced PFBF and AUC 5 by 29% (P<0.05) and 39% (P<0.0005) respectively in controls, but had no significant effect on these parameters in DM subjects. Infusion of ASA reduced FBF, PFBF and AUC 5 in both DM (P<0.05) and control (P<0.05) subjects, but the magnitude of this reduction was greater in DM than in control subjects (ANOVA, P<0.05), even when differences in resting FBF were accounted for. Indeed, ASA eliminated the differences in FBF, PFBF and AUC 5 between DM and control subjects. Thus increased release of vasodilator prostanoids, rather than of NO, appears to account for skeletal muscle hyperaemia in Type I diabetes. PMID- 11052919 TI - Comparison of pulse-wave velocity in different aortic regions in relation to the extent and severity of atherosclerosis between young and older Kurosawa and Kusanagi-hypercholesterolemic (KHC) rabbits. AB - The present study was performed to investigate the effects of the development of atherosclerosis on foot-to-foot pulse-wave velocity (PWV) from the ascending aorta to different positions along the aorta in Kurosawa and Kusanagi Hypercholesterolemic (KHC) rabbits aged 10-12 and 22-24 months old, in relation to the percentage fractional lesioned area (PFLA) in different aortic regions through which the pulse wave travels, as well as the rheological and pathohistological properties of the aortic wall. PWV, when measured in the KHC rabbit from the ascending aorta to each aortic position, showed the highest value on passage through the aortic arch, decreased with conduction to the distal thoracic aorta, reached the minimal value on passage to the distal thoracic aorta or to the middle abdominal aorta in the 10-12- and 22-24-month-old animals respectively, and increased gradually on conduction to the iliac artery. PWV at all aortic regions examined was significantly greater in the 22-24-month-old than in the 10-12-month-old KHC rabbits. PFLA, when measured in the aortic region from the ascending aorta to each aortic position, was maximal in the aortic arch and decreased gradually towards the peripheral aorta in both age groups. PFLA in the 22-24-month-old group was significantly greater than that in the 10-12-month-old group in all aortic regions examined. The atherosclerotic aortic wall showed a significantly lower elastic modulus in the young KHC rabbits compared with age matched normal rabbits. A significantly higher elastic modulus was observed in the older KHC rabbits compared with that in the younger animals of both strains, associated with the progression of sclerosis. These findings indicate that the increase in PWV is due mainly to an increase with aging in the extent and severity of atherosclerosis in vessels in which the pulse wave travels. PMID- 11052920 TI - Effect of ciprofloxacin on the activation of the transcription factors nuclear factor kappaB, activator protein-1 and nuclear factor-interleukin-6, and interleukin-6 and interleukin-8 mRNA expression in a human endothelial cell line. AB - Quinolone antibiotics such as ciprofloxacin modify immune and inflammatory responses in some cells. We have shown previously that ciprofloxacin decreases the accumulation of interleukin (IL)-6 protein from a human endothelial cell line, whilst IL-8 protein production was increased. It is not known whether this occurs through effects on transcription and mRNA expression. We therefore investigated the effect of ciprofloxacin on mRNA for IL-6 and IL-8, and on three transcription factors known to be involved in the regulation of these cytokines. We investigated the effect of ciprofloxacin on tumour necrosis factor alpha- and IL-1beta-mediated activation of the transcription factors nuclear factor kappaB (NFkappaB), activator protein-1 (AP-1) and nuclear factor IL-6 (NF-IL-6) using an electrophoretic mobility shift assay, and the effect on expression of mRNA for IL 6 and IL-8 by reverse transcriptase-PCR in the EAhy926 endothelial cell line. Ciprofloxacin decreased IL-6 mRNA (P<0.05) and increased IL-8 mRNA (P<0.05) expression. Ciprofloxacin did not modulate activation of NFkappaB or AP-1. However, NF-IL-6 binding was decreased in the presence of 100 microg/ml ciprofloxacin (P<0.05). The study shows that ciprofloxacin-mediated decreased IL 6 release by a human endothelial cell line is reflected by decreased mRNA expression and decreased NF-IL-6 but not NFkappaB or AP-1 activation. Increased IL-8 mRNA in response to ciprofloxacin was not reflected by altered transcription factor activation and may represent increased mRNA stability. PMID- 11052921 TI - Left atrial thrombin generation and prothrombin fragment 1+2. PMID- 11052922 TI - Dexamethasone and pentastarch produce additive attenuation of ischaemia/reperfusion lung injury. AB - The choice of an intravenous solution for the attenuation of ischaemia/reperfusion (I/R) lung injury is still a difficult one. Although 10% (w/v) pentastarch has been used in ICU settings, its use in I/R lung injury has not been well explored. We hypothesized that this synthetic colloid substance, which maintains colloid osmotic pressure and potentially 'seals' capillary leaks, in combination with an anti-inflammatory agent (i.e. dexamethasone), would ameliorate I/R lung injury. After 60 min of lung ischaemia in an isolated rat lung model, lungs were reperfused for 60 min in a closed circulating system with one of the following solutions: (1) NS (0.9% normal saline), (2) NS+Dex (dexamethasone), (3) NS+Penta (pentastarch), or (4) NS+Penta+Dex. Haemodynamic changes, lung weight gain (LWG), capillary filtration coefficient (K(fc)) and lung pathology were analysed. Results showed significantly lower values of K(fc) and LWG in pentastarch- or dexamethasone-perfused groups as compared with those in the NS group. Dexamethasone as an additive to NS+Penta further decreased K(fc) and LWG. Histopathological studies showed similar decreases in injury profiles. We conclude that reperfusion with dexamethasone and pentastarch can attenuate I/R lung injury, and that dexamethasone and pentastarch have additive effects. Our data thus suggest that the combination of a colloid substance with 'sealing effects' and an anti-inflammatory agent may provide a better reperfusion solution for patients with I/R lung injury or for lungs stored for transplant. PMID- 11052923 TI - Effects of the Japanese herbal medicine 'Inchinko-to' (TJ-135) on concanavalin A induced hepatitis in mice. AB - Inchinko-to (TJ-135) is a herbal medicine consisting of three kinds of crude drugs, and in Japan it is administered mainly to patients with cholestasis. The present study evaluated the effects of TJ-135 on concanavalin A (con A)-induced hepatitis in mice in vivo and con A-induced cytokine production in vitro. When mice were pretreated with oral TJ-135 for 1 week before intravenous con A injection, the activities of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were significantly decreased 8 h after con A administration (-82%, -96% and -66% respectively). In histological investigations, sub-massive hepatic necrosis accompanying inflammatory cell infiltration was not observed in mice pretreated with TJ-135. Serum levels of interleukin-12 (IL-12), interferon-gamma (IFN-gamma) and IL-2 were significantly lower in mice pretreated with TJ-135 compared with controls, while IL-10 levels were higher in these mice. Intrasplenic IL-12 levels were significantly lower in mice pretreated with TJ-135, while intrasplenic IL-10 levels were higher in these mice. In vitro, IL-10 production by splenocytes was increased by the addition of TJ-135 to the culture medium, whereas the production of IL-12 and IFN-gamma was inhibited. These results suggest that con A-induced hepatitis was ameliorated by pretreatment with TJ-135. With regard to the mechanism of these effects of TJ-135, we speculate that TJ-135 inhibits the production of inflammatory cytokine and enhances the production of anti inflammatory cytokines. Therefore administration of TJ-135 may be useful in patients with severe acute hepatitis accompanying cholestasis or in those with autoimmune hepatitis. PMID- 11052924 TI - Pharmacokinetics of retinyl palmitate and retinol after intramuscular retinyl palmitate administration in severe malaria. AB - Retinol (vitamin A alcohol) is an accepted adjunctive treatment in infections such as measles. There is also indirect evidence from in vitro, animal and human studies that retinol supplementation may be beneficial in severe malaria. There have, however, been no studies that have examined the pharmacokinetics of acute retinol supplementation in severe illness. To establish whether mobilization of intramuscular retinyl palmitate (RP) and its availability as retinol are adequate in complicated falciparum malaria, we administered a single dose of 400000 i.u. of RP to six Vietnamese adults with severe malaria. Another 28 patients were not given RP. All patients had blood samples taken over 96 h for RP and retinol assay using HPLC, and received conventional anti-malarial and supportive therapy. Admission serum retinol concentrations were below the lower limit of the reference range (<1.0 micromol/l) in 74% of the 34 patients. In supplemented patients, analysis of serum RP between 0 and 96 h using a multi-compartmental model revealed a median (range) delay in mobilization of 6.9 h (0.7-15.1 h), a bioavailability of 55% (19-100%) and an elimination half-life of 13.5 h (4.2-23.7 h). The area under the serum retinol curve expressed as an absolute or percentage change from baseline was greater in supplemented than in unsupplemented patients (P<0.05). The separation in median serum retinol concentrations in the two groups was maximal at 48 h. The model-derived retinol half-life [1.5 (0.7-15.8) h] suggested rapid uptake, metabolism and/or excretion. In conclusion, there is variable RP bioavailability in severe malaria, but a significant if delayed increase in serum retinol over that associated with recovery from the infection. In severe infections, RP supplementation appears simple, well tolerated and of potential benefit once anti-microbial and supportive therapy have been established. PMID- 11052925 TI - Characterization of an in vitro model for the study of the short and prolonged effects of myocardial ischaemia and reperfusion in man. AB - The mechanisms underlying myocardial ischaemia and reperfusion-induced injury have been investigated, mainly by using animal experimental preparations in vitro and in vivo, but little is known of the process in human myocardium. The present studies characterize an in vitro model using human myocardium for the study of early and delayed effects of ischaemia and reperfusion. The right atrial appendage was manually sliced and incubated in buffer through which was bubbled O(2)/CO(2) (19:1, v/v) for various time periods. Lactate dehydrogenase (LDH) leakage, 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl-2H-tetrazolium bromide (MTT) reduction, oxygen consumption, nucleotide levels and tissue morphology were all investigated as markers of myocardial injury. The specimens remained stable and viable up to 24 h, but had significantly deteriorated by 48 h. The preparation responded to ischaemia in a time-related manner. Tissue viability was reduced by 25% after 30 min ischaemia, declined to 60% after 60 min ischaemia and to 75% after 120 min ischaemia. Interestingly, the tissue was more susceptible when ischaemia was induced after 24 h of aerobic incubation. The effects of the duration of reperfusion were investigated after a fixed 60 min ischaemic insult. The results of LDH leakage suggest that reperfusion injury is mainly sustained within the first 2 h of reperfusion. However, the results of MTT reduction show that there is a progressive decrease in tissue viability over the 24 h reperfusion period, possibly reflecting the occurrence of tissue necrosis and apoptosis at different reperfusion times. In conclusion, the data provide evidence that the incubation of human atrial tissue in vitro is stable, and slices are viable for at least 24 h, which permits the study of early and delayed consequences of ischaemia and reperfusion in the human myocardium. PMID- 11052926 TI - Mesenteric hyporesponsiveness in cirrhotic rats with ascites: role of cGMP and K+ channels. AB - The mechanisms that mediate hyporesponsiveness to vasoconstrictors in liver cirrhosis are not completely established. In the present study we have explored the role of NO and potassium channels by studying the pressor response to methoxamine in rats with carbon tetrachloride-induced cirrhosis with ascites. Experiments were performed in the isolated and perfused mesenteric arterial bed of control rats and of cirrhotic rats with ascites. Pressor responses to methoxamine, an alpha-adrenergic agonist, were analysed under basal conditions, after inhibition of guanylate cyclase with Methylene Blue (MB; 10 microM), after inhibition of NO synthesis with N(G)-nitro-L-arginine (L-NNA; 100 microM) and after blockade of potassium channels with tetraethylammonium (TEA; 3 mM). Compared with those from controls, preparations from cirrhotic rats showed a lower pressor response to methoxamine (maximum: controls, 114.4+/-6.8 mmHg; cirrhotic rats, 74.7+/-7.3 mmHg). Pretreatment with MB or L-NNA increased the responses in both groups, but without correcting the lower than normal response of the cirrhotic rats. Pretreatment with TEA alone did not modify the responses as compared with the untreated groups. Pretreatment with TEA plus MB or TEA plus L-NNA also potentiated the responses, and the responses of the cirrhotic animals were greater than those of the groups treated with MB or L-NNA alone. However, no treatment completely normalized the lower response of the mesenteries from cirrhotic animals, suggesting that factors other than NO and potassium channels also participate, although to a lesser degree, in the lower pressor response of the mesenteric arterial bed of animals with cirrhosis. These results confirm that NO and potassium channels are important mediators of the lower vascular pressor response of the mesenteric bed of cirrhotic rats with ascites. This effect seems to be mediated by the NO-dependent formation of cGMP and by the NO-dependent and independent activation of potassium channels. PMID- 11052927 TI - Plasma bradykinin levels in human chronic congestive heart failure. AB - Induction of congestive heart failure by high-frequency pacing has been reported to increase plasma levels of immunoreactive kinins in dogs. In the present study, we evaluated plasma bradykinin levels in human heart failure. Utilizing a recently developed method, we specifically measured plasma levels of bradykinin (1-9) nonapeptide in 21 patients with chronic congestive heart failure [New York Heart Association (NYHA) stages III and IV). At the same time, we measured plasma atrial natriuretic peptide levels and plasma renin activity, and, as a marker of inflammation, plasma levels of tumour necrosis factor. In addition, 18 healthy subjects matched for gender and age served as normal controls. Plasma bradykinin concentrations were not higher in patients with chronic congestive heart failure (median 2.1 fmol/ml) than in healthy subjects (2.6 fmol/ml). In contrast, plasma atrial natriuretic peptide levels were clearly higher (patients, 63 fmol/ml; controls, 24 fmol/ml; P<0.0001), despite diuretic treatment and in the presence of high plasma renin activity (patients, 13.0 ng x h(-1) x ml(-1); controls, 0.3 ng x h(-1) x ml(-1); P<0.0001). Tumour necrosis factor was elevated in heart failure patients in NYHA class IV only (27 pg/ml, compared with 21 pg/ml in controls; P=0.013). Bradykinin, atrial natriuretic peptide and plasma renin activity levels were not correlated with the severity of the disease, as assessed by NYHA classification. These results indicate that a rather selective cytokine activation, without concomitant stimulation of the kallikrein-kinin system, occurs in human chronic congestive heart failure. PMID- 11052928 TI - Circulating C-type natriuretic peptide is increased in orthotopic cardiac transplant recipients and associated with cardiac allograft vasculopathy. AB - C-type natriuretic peptide (CNP) is a potent, endothelial-derived relaxant and growth-inhibitory factor. Accelerated vascular disease is an important cause of morbidity in cardiac transplant recipients, and endothelial dysfunction is now well recognized in patients with cardiovascular disease. CNP has not previously been investigated following cardiac transplantation. We therefore studied plasma levels of immunoreactive CNP in patients early and late after heart transplantation, compared with levels in healthy subjects. We measured CNP in extracted human plasma using an antibody against human CNP-(1-22). CNP levels were significantly elevated in 13 cardiac recipients 2 weeks post-transplant [2.64+/-0.26 pmol/l (mean+/-S.E.M.)] compared with those in the normal healthy subjects (0.62+/-0.04 pmol/l; n=20, P<0.001). Plasma levels of CNP were also significantly elevated in a second group of established cardiac transplant recipients (1.15+/-0.07 pmol/l; n=46) studied 1-13 years post-transplant when compared with the healthy subjects (P<0.001). In the group studied later after transplantation, CNP levels were significantly associated with systolic blood pressure (P<0.05) and were higher in patients with angiographic post-transplant coronary artery disease (P=0.032). In conclusion, these findings clearly demonstrate that CNP is elevated soon after cardiac transplantation and remains raised in patients even several years post-transplant. CNP may be important as a circulating or local hormone involved in vascular contractile function and in the pathophysiology of cardiac allograft vasculopathy following heart transplantation. PMID- 11052929 TI - Diversity and dynamics of dendritic signaling. AB - Communication between neurons in the brain occurs primarily through synapses made onto elaborate treelike structures called dendrites. New electrical and optical recording techniques have led to tremendous advances in our understanding of how dendrites contribute to neuronal computation in the mammalian brain. The varied morphology and electrical and chemical properties of dendrites enable a spectrum of local and long-range signaling, defining the input-output relationship of neurons and the rules for induction of synaptic plasticity. In this way, diversity in dendritic signaling allows individual neurons to carry out specialized functions within their respective networks. PMID- 11052930 TI - Untangling dendrites with quantitative models. AB - Our understanding of the function of dendrites has been greatly enriched by an inspiring dialogue between theory and experiments. Rather than functionally ignoring dendrites, representing neurons as single summing points, we have realized that dendrites are electrically and chemically distributed nonlinear units and that this has important consequences for interpreting experimental data and for the role of neurons in information processing. Here, we examine the route to unraveling some of the enigmas of dendrites and highlight the main insights that have been gained. Future directions are discussed that will enable theory and models to keep shedding light on dendrites, where the most fundamental input output adaptive processes take place. PMID- 11052931 TI - Signal-processing machines at the postsynaptic density. AB - Dendrites of individual neurons in the vertebrate central nervous system are contacted by thousands of synaptic terminals relaying information about the environment. The postsynaptic membrane at each synaptic terminal is the first place where information is processed as it converges on the dendrite. At the postsynaptic membrane of excitatory synapses, neurotransmitter receptors are attached to large protein "signaling machines" that delicately regulate the strength of synaptic transmission. These machines are visible in the electron microscope and are called the postsynaptic density. By changing synaptic strength in response to neural activity, the postsynaptic density contributes to information processing and the formation of memories. PMID- 11052932 TI - Actin-based plasticity in dendritic spines. AB - The central nervous system functions primarily to convert patterns of activity in sensory receptors into patterns of muscle activity that constitute appropriate behavior. At the anatomical level this requires two complementary processes: a set of genetically encoded rules for building the basic network of connections, and a mechanism for subsequently fine tuning these connections on the basis of experience. Identifying the locus and mechanism of these structural changes has long been among neurobiology's major objectives. Evidence has accumulated implicating a particular class of contacts, excitatory synapses made onto dendritic spines, as the sites where connective plasticity occurs. New developments in light microscopy allow changes in spine morphology to be directly visualized in living neurons and suggest that a common mechanism, based on dynamic actin filaments, is involved in both the formation of dendritic spines during development and their structural plasticity at mature synapses. PMID- 11052934 TI - Quantum superposition of macroscopic persistent-current states. AB - Microwave spectroscopy experiments have been performed on two quantum levels of a macroscopic superconducting loop with three Josephson junctions. Level repulsion of the ground state and first excited state is found where two classical persistent-current states with opposite polarity are degenerate, indicating symmetric and antisymmetric quantum superpositions of macroscopic states. The two classical states have persistent currents of 0.5 microampere and correspond to the center-of-mass motion of millions of Cooper pairs. PMID- 11052933 TI - Neurodegeneration prevented by lentiviral vector delivery of GDNF in primate models of Parkinson's disease. AB - Lentiviral delivery of glial cell line-derived neurotrophic factor (lenti-GDNF) was tested for its trophic effects upon degenerating nigrostriatal neurons in nonhuman primate models of Parkinson's disease (PD). We injected lenti-GDNF into the striatum and substantia nigra of nonlesioned aged rhesus monkeys or young adult rhesus monkeys treated 1 week prior with 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). Extensive GDNF expression with anterograde and retrograde transport was seen in all animals. In aged monkeys, lenti-GDNF augmented dopaminergic function. In MPTP-treated monkeys, lenti-GDNF reversed functional deficits and completely prevented nigrostriatal degeneration. Additionally, lenti-GDNF injections to intact rhesus monkeys revealed long-term gene expression (8 months). In MPTP-treated monkeys, lenti-GDNF treatment reversed motor deficits in a hand-reach task. These data indicate that GDNF delivery using a lentiviral vector system can prevent nigrostriatal degeneration and induce regeneration in primate models of PD and might be a viable therapeutic strategy for PD patients. PMID- 11052935 TI - Triple vortex ring structure in superfluid helium II. AB - Superfluids such as helium II consist of two interpenetrating fluids: the normal fluid and the superfluid. The helium II vortex ring has generally been considered merely as a superfluid object, neglecting any associated motion of the normal fluid. We report a three-dimensional calculation of the coupled motion of the normal-fluid and superfluid components, which shows that the helium II vortex ring consists of a superfluid vortex ring accompanied by two coaxial normal-fluid vortex rings of opposite polarity. The three vortex rings form a coherent, dissipative structure. PMID- 11052936 TI - Phase evolution in a Kondo-correlated system. AB - We measured the phase evolution of electrons as they traverse a quantum dot (QD) formed in a two-dimensional electron gas that serves as a localized spin. The traversal phase, determined by embedding the QD in a double path electron interferometer and measuring the quantum interference of the electron wave functions manifested by conductance oscillation as a function of a weak magnetic field, evolved by pi radians, a range twice as large as theoretically predicted. As the correlation weakened, a gradual transition to the familiar phase evolution of a QD was observed. The specific phase evolution observed is highly sensitive to the onset of Kondo correlation, possibly serving as an alternative fingerprint of the Kondo effect. PMID- 11052937 TI - Moissanite: a window for high-pressure experiments. AB - We achieved a pressure of 52.1 gigapascals with moissanite anvils, which have optical, thermal, electric, magnetic, and x-ray properties that rival those of diamond. The mode-softening of D(2)O toward the pressure-induced hydrogen bond symmetrization and the Raman shifts of diamond under hydrostatic and nonhydrostatic compressions were studied with moissanite anvils in the spectral regions normally obscured by diamond anvils. Moissanite anvil cells allow maximum sample volumes 1000 times larger than those allowed by diamond anvil cells and may enable the next level of advancement in high-pressure experiments. PMID- 11052938 TI - Forster energy transfer in an optical microcavity. AB - By studying the transfer of excitation energy between dye molecules confined within an optical microcavity, we demonstrate experimentally that Forster energy transfer is influenced by the local photonic mode density. Locating donor and acceptor molecules at well-defined positions allows the transfer rate to be determined as a function of both mutual separation and cavity length. The results show that the Forster transfer rate depends linearly on the donor emission rate and hence photonic mode density, providing the potential to control energy transfer by modification of the optical environment. PMID- 11052939 TI - Hurricane disturbance and tropical tree species diversity. AB - The debate over the maintenance of high diversity of tree species in tropical forests centers on the role of tree-fall gaps as a primary source of disturbance. Using a 10-year data series accumulated since Hurricane Joan struck the Caribbean coast of Nicaragua in 1988, we examined the pattern of species accumulation over time and with increased sampling of individuals. Our analysis shows that the pattern after a hurricane differs from the pattern after a simple tree-fall disturbance, and we conclude that pioneers are limited in large disturbances and thus do not suppress other species the way they do in smaller disturbances. PMID- 11052940 TI - A low temperature transfer of ALH84001 from Mars to Earth. AB - The ejection of material from Mars is thought to be caused by large impacts that would heat much of the ejecta to high temperatures. Images of the magnetic field of martian meteorite ALH84001 reveal a spatially heterogeneous pattern of magnetization associated with fractures and rock fragments. Heating the meteorite to 40 degrees C reduces the intensity of some magnetic features, indicating that the interior of the rock has not been above this temperature since before its ejection from the surface of Mars. Because this temperature cannot sterilize most bacteria or eukarya, these data support the hypothesis that meteorites could transfer life between planets in the solar system. PMID- 11052941 TI - Impacts of climatic change and fishing on Pacific salmon abundance over the past 300 years. AB - The effects of climate variability on Pacific salmon abundance are uncertain because historical records are short and are complicated by commercial harvesting and habitat alteration. We use lake sediment records of delta15N and biological indicators to reconstruct sockeye salmon abundance in the Bristol Bay and Kodiak Island regions of Alaska over the past 300 years. Marked shifts in populations occurred over decades during this period, and some pronounced changes appear to be related to climatic change. Variations in salmon returns due to climate or harvesting can have strong impacts on sockeye nursery lake productivity in systems where adult salmon carcasses are important nutrient sources. PMID- 11052942 TI - Antiphase oscillation of the left and right suprachiasmatic nuclei. AB - An unusual property of the circadian timekeeping systems of animals is rhythm "splitting," in which a single daily period of physical activity (usually measured as wheel running) dissociates into two stably coupled components about 12 hours apart; this behavior has been ascribed to a clock composed of two circadian oscillators cycling in antiphase. We analyzed gene expression in the hypothalamic circadian clock, the suprachiasmatic nucleus (SCN), of behaviorally "split" hamsters housed in constant light. The results show that the two oscillators underlying the split condition correspond to the left and right sides of the bilaterally paired SCN. PMID- 11052943 TI - Integration of multiple signals through cooperative regulation of the N-WASP Arp2/3 complex. AB - The protein N-WASP [a homolog to the Wiskott-Aldrich syndrome protein (WASP)] regulates actin polymerization by stimulating the actin-nucleating activity of the actin-related protein 2/3 (Arp2/3) complex. N-WASP is tightly regulated by multiple signals: Only costimulation by Cdc42 and phosphatidylinositol (4,5) bisphosphate (PIP2) yields potent polymerization. We found that regulation requires N-WASP's constitutively active output domain (VCA) and two regulatory domains: a Cdc42-binding domain and a previously undescribed PIP(2)-binding domain. In the absence of stimuli, the regulatory modules together hold the VCA Arp2/3 complex in an inactive "closed" conformation. In this state, both the Cdc42- and PIP2-binding sites are masked. Binding of either input destabilizes the closed state and enhances binding of the other input. This cooperative activation mechanism shows how combinations of simple binding domains can be used to integrate and amplify coincident signals. PMID- 11052944 TI - Direct coupling between meiotic DNA replication and recombination initiation. AB - During meiosis in Saccharomyces cerevisiae, DNA replication occurs 1. 5 to 2 hours before recombination initiates by DNA double-strand break formation. We show that replication and recombination initiation are directly linked. Blocking meiotic replication prevented double-strand break formation in a replication checkpoint-independent manner, and delaying replication of a chromosome segment specifically delayed break formation in that segment. Consequently, the time between replication and break formation was held constant in all regions. We suggest that double-strand break formation occurs as part of a process initiated by DNA replication, which thus determines when meiotic recombination initiates on a regional rather than a cell-wide basis. PMID- 11052945 TI - Genomic analysis of gene expression in C. elegans. AB - Until now, genome-wide transcriptional profiling has been limited to single-cell organisms. The nematode Caenorhabditis elegans is a well-characterized metazoan in which the expression of all genes can be monitored by oligonucleotide arrays. We used such arrays to quantitate the expression of C. elegans genes throughout the development of this organism. The results provide an estimate of the number of expressed genes in the nematode, reveal relations between gene function and gene expression that can guide analysis of uncharacterized worm genes, and demonstrate a shift in expression from evolutionarily conserved genes to worm specific genes over the course of development. PMID- 11052946 TI - Song replay during sleep and computational rules for sensorimotor vocal learning. AB - Songbirds learn a correspondence between vocal-motor output and auditory feedback during development. For neurons in a motor cortex analog of adult zebra finches, we show that the timing and structure of activity elicited by the playback of song during sleep matches activity during daytime singing. The motor activity leads syllables, and the matching sensory response depends on a sequence of typically up to three of the preceding syllables. Thus, sensorimotor correspondence is reflected in temporally precise activity patterns of single neurons that use long sensory memories to predict syllable sequences. Additionally, "spontaneous" activity of these neurons during sleep matches their sensorimotor activity, a form of song "replay." These data suggest a model whereby sensorimotor correspondences are stored during singing but do not modify behavior, and off-line comparison (e.g., during sleep) of rehearsed motor output and predicted sensory feedback is used to adaptively shape motor output. PMID- 11052947 TI - Structure of murine CTLA-4 and its role in modulating T cell responsiveness. AB - The effective regulation of T cell responses is dependent on opposing signals transmitted through two related cell-surface receptors, CD28 and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4). Dimerization of CTLA-4 is required for the formation of high-avidity complexes with B7 ligands and for transmission of signals that attenuate T cell activation. We determined the crystal structure of the extracellular portion of CTLA-4 to 2.0 angstrom resolution. CTLA-4 belongs to the immunoglobulin superfamily and displays a strand topology similar to Valpha domains, with an unusual mode of dimerization that places the B7 binding sites distal to the dimerization interface. This organization allows each CTLA-4 dimer to bind two bivalent B7 molecules and suggests that a periodic arrangement of these components within the immunological synapse may contribute to the regulation of T cell responsiveness. PMID- 11052948 TI - Muscle glycogen content affects insulin-stimulated glucose transport and protein kinase B activity. AB - We investigated the possible regulatory role of glycogen in insulin-stimulated glucose transport and insulin signaling in skeletal muscle. Rats were preconditioned to obtain low (LG), normal, or high (HG) muscle glycogen content, and perfused isolated hindlimbs were exposed to 0, 100, or 10,000 microU/ml insulin. In the fast-twitch white gastrocnemius, insulin-stimulated glucose transport was significantly higher in LG compared with HG. This difference was less pronounced in the mixed-fiber red gastrocnemius and was absent in the slow twitch soleus. In the white gastrocnemius, insulin activation of insulin receptor tyrosine kinase and phosphoinositide 3-kinase was unaffected by glycogen levels, whereas protein kinase B activity was significantly higher in LG compared with HG. In additional incubation experiments on fast-twitch epitrochlearis muscles, insulin-stimulated cell surface GLUT-4 content was significantly higher in LG compared with HG. The data indicate that, in fast-twitch muscle, the effect of insulin on glucose transport and cell surface GLUT-4 content is modulated by glycogen content, which does not involve initial but possibly more downstream signaling events. PMID- 11052949 TI - Reduced GLP-1 and insulin responses and glucose intolerance after gastric glucose in GRP receptor-deleted mice. AB - By applying a newly developed ELISA technique for determining biologically active intact glucagon-like peptide [GLP-1, GLP-1-(7-36)amide] in mouse, plasma baseline GLP-1 in normal NMRI mice was found to be normally distributed (4.5 +/- 0.3 pmol/l; n = 72). In anesthetized mice, gastric glucose (50 or 150 mg) increased plasma GLP-1 levels two- to threefold (P < 0.01). The simultaneous increase in plasma insulin correlated to the 10-min GLP-1 levels (r = 0.36, P < 0.001; n = 12). C57BL/6J mice deleted of the gastrin-releasing peptide (GRP) receptor by genetic targeting had impaired glucose tolerance (P = 0.030) and reduced early (10 min) insulin response (P = 0.044) to gastric glucose compared with wild-type controls. Also, the GLP-1 response to gastric glucose was significantly lower in the GRP receptor-deleted mice than in the controls (P = 0.045). In conclusion, this study has shown that 1) plasma levels of intact GLP-1 increase dose dependently on gastric glucose challenge in correlation with increased insulin levels in mice, and 2) intact GRP receptors are required for normal GLP-1 and insulin responses and glucose tolerance after gastric glucose in mice. PMID- 11052950 TI - Norepinephrine-induced sustained inward current in brown fat cells: alpha(1) mediated by nonselective cation channels. AB - The nature of the sustained norepinephrine-induced depolarization in brown fat cells was examined by patch-clamp techniques. Norepinephrine (NE) stimulation led to a whole cell current response consisting of two phases: a first inward current, lasting for only 1 min, and a sustained inward current, lasting as long as the adrenergic stimulation was maintained. The nature of the sustained current was here investigated. It could be induced by the alpha(1)-agonist cirazoline but not by the beta(3)-agonist CGP-12177A. Reduction of extracellular Cl(-) concentration had no effect, but omission of extracellular Ca(2+) or Na(+) totally eliminated it. When unstimulated cells were studied in the cell-attached mode, some activity of approximately 30 pS nonselective cation channels was observed. NE perfusion led to a 10-fold increase in their open probability (from approximately 0.002 to approximately 0.017), which persisted as long as the perfusion was maintained. The activation was much stronger with the alpha(1) agonist phenylephrine than with the beta(3)-agonist CGP-12177A, and with the Ca(2+) ionophore A-23187 than with the adenylyl cyclase activator forskolin. We conclude that the sustained inward current was due to activation of approximately 30 pS nonselective cation channels via alpha(1)-adrenergic receptors and that the effect may be mediated via an increase in intracellular free Ca(2+) concentration. PMID- 11052951 TI - Whole body protein kinetics in women: effect of pregnancy and IDDM during anabolic stimulation. AB - The effects of pregnancy and type 1 diabetes [insulin-dependent diabetes mellitus (IDDM)] on protein metabolism are still uncertain. Therefore, six normal and five IDDM women were studied during and after pregnancy, using [(13)C]leucine and [(2)H(5)]phenylalanine with a hyperinsulinemic-euglycemic clamp and amino acid infusion. Fasting total plasma amino acids were lower in pregnancy in normal but not IDDM women (2,631 +/- 427 vs. 2,057 +/- 471 and 2,523 +/- 430 vs. 2,500 +/- 440 micromol/l, respectively). Whole body protein breakdown (leucine) increased in pregnancy [change in normal (delta N) and IDDM women (delta D) 0.59 +/- 0.40 and 0.48 +/- 0.26 g. kg(-1). day(-1), both P < 0.001], whereas reductions in protein breakdown due to insulin/amino acids (delta N -0.57 +/- 0.19, delta D 0.58 +/- 0.20 g. kg(-1). day(-1), both P < 0.001) were unaffected by pregnancy. Protein breakdown in IDDM women was not higher than normal, and neither pregnancy nor type 1 diabetes altered the insulin sensitivity of amino acid turnover. Nonoxidized leucine disposal (protein synthesis) increased in pregnancy (delta N 0.67 +/- 0.45, delta D 0.64 +/- 0.34 g. kg(-1). day(-1), both P < 0.001). Pregnancy reduced the response of phenylalanine hydroxylation to insulin/amino acids in both groups (delta N -1.14 +/- 0.74, delta D -1. 12 +/- 0.77 g. kg(-1). day(-1), both P < 0.05). These alterations may enable amino acid conservation for protein synthesis and accretion in late pregnancy. Well-controlled type 1 diabetes caused no abnormalities in the regulation of basal or stimulated protein metabolism. PMID- 11052952 TI - Growth hormone enhances effects of endurance training on oxidative muscle metabolism in elderly women. AB - The present study investigated whether recombinant human (rh) growth hormone (GH) combined with endurance training would have a larger effect on oxidative capacity, metabolism, and body fat than endurance training alone. Sixteen healthy, elderly women, aged 75 yr, performed closely monitored endurance training on a cycle ergometer over 12 wk. rhGH was given in a randomized, double blinded, placebo-controlled design in addition to the training program. GH administration resulted in a doubling of serum insulin-like growth factor I levels. With endurance training, peak oxygen uptake increased by approximately 18% in both groups, whereas the marked increase in muscle citrate synthase activity was 50% larger in the GH group compared with the placebo group. In addition, only the GH group revealed an increase in muscle L-3-hydroxyacyl-CoA dehydrogenase activity. Body weight remained unchanged in both groups, but the GH group showed significant changes in body composition with a decrease in fat mass and an increase in lean body mass. Twenty-four-hour indirect calorimetry performed in four subjects showed a marked increase in energy expenditure with increased relative and absolute fat combustion in the two subjects receiving rhGH. In conclusion, rhGH adds to the effects of endurance training on muscle oxidative enzymes and causes a reduction in body fat in elderly women. PMID- 11052953 TI - Determinants of glucose toxicity and its reversibility in the pancreatic islet beta-cell line, HIT-T15. AB - HIT-T15 cells, a clonal beta-cell line, were cultured and passaged weekly for 6 mo in RPMI 1640 media containing various concentrations of glucose. Insulin content decreased in the intermediate- and late-passage cells as a continuous rather than a threshold glucose concentration effect. In a second series of experiments, cells were grown in media containing either 0.8 or 16.0 mM glucose from passages 76 through 105. Subcultures of passages 86, 92, and 99 that had been grown in media containing 16.0 mM glucose were switched to media containing 0.8 mM glucose and also carried forward to passage 105. Dramatic increases in insulin content and secretion and insulin gene expression were observed when the switches were made at passages 86 and 92 but not when the switch was made at passage 99. These findings suggest that glucose toxicity of insulin-secreting cells is a continuous rather than a threshold function of glucose concentration and that the shorter the period of antecedent glucose toxicity, the more likely that full recovery of cell function will occur. PMID- 11052954 TI - Glucose-stimulated insulin secretion suppresses hepatic triglyceride-rich lipoprotein and apoB production. AB - The current study assessed in vivo the effect of insulin on triglyceride-rich lipoprotein (TRL) production by rat liver. Hepatic triglyceride and apolipoprotein B (apoB) production were measured in anesthetized, fasted rats injected intravenously with Triton WR-1339 (400 mg/kg). After intravascular catabolism was blocked by detergent treatment, glucose (500 mg/kg) was injected to elicit insulin secretion, and serum triglyceride and apoB accumulation were monitored over the next 3 h. In glucose-injected rats, triglyceride secretion averaged 22.5 +/- 2.1 microg.ml(-1).min(-1), which was significantly less by 30% than that observed in saline-injected rats, which averaged 32.1 +/- 1.4 microg.ml(-1).min(-1). ApoB secretion was also significantly reduced by 66% in glucose-injected rats. ApoB immunoblotting indicated that both B100 and B48 production were significantly reduced after glucose injection. Results support the conclusion that insulin acts in vivo to suppress hepatic very low density lipoprotein (VLDL) triglyceride and apoB secretion and strengthen the concept of a regulatory role for insulin in VLDL metabolism postprandially. PMID- 11052955 TI - Responses of adipose tissue lipoprotein lipase to weight loss affect lipid levels and weight regain in women. AB - This study determines whether changes in abdominal (ABD) and gluteal (GLT) adipose tissue lipoprotein lipase (LPL) activity in response to a 6-mo weight loss intervention, comprised of a hypocaloric diet and low-intensity walking, affect changes in body composition, fat distribution, lipid metabolism, and the magnitude of weight regain in 36 obese postmenopausal women. Average adipose tissue LPL activity did not change with an average 5.6-kg weight loss, but changes in LPL activity were inversely related to baseline LPL activity (ABD: r = -0.60, GLT: r = -0.48; P < 0.01). The loss of abdominal body fat and decreases in total and low-density lipoprotein cholesterol were greater in women whose adipose tissue LPL activity decreased with weight loss despite a similar loss of total body weight and fat mass. Moreover, weight regain after a 6-mo follow-up was less in women whose adipose tissue LPL activity decreased than in women whose LPL increased (ABD: 0.9 +/- 0.5 vs. 2.8 +/- 0.6 kg, P < 0.05; GLT: 0.2 +/- 0.5 vs. 2.8 +/- 0.5 kg, P < 0.01). These results suggest that a reduction in adipose tissue LPL activity with weight loss is associated with improvements in lipid metabolic risk factors with weight loss and with diminished weight regain in postmenopausal women. PMID- 11052956 TI - Prior exercise and postprandial substrate extraction across the human leg. AB - Prior exercise decreases postprandial plasma triacylglycerol (TG) concentrations, possibly through changes to skeletal muscle TG extraction. We measured postprandial substrate extraction across the leg in eight normolipidemic men aged 21-46 yr. On the afternoon preceding one trial, subjects ran for 2 h at 64 +/- 1% of maximal oxygen uptake (exercise); before the control trial, subjects had refrained from exercise. Samples of femoral arterial and venous blood were obtained, and leg blood flow was measured in the fasting state and for 6 h after a meal (1.2 g fat, 1.2 g carbohydrate/kg body mass). Prior exercise increased time averaged postprandial TG clearance across the leg (total TG: control, 0.079 +/- 0.014 ml.100 ml tissue(-1).min(-1) ; exercise, 0.158 +/- 0.023 ml.100 ml tissue(-1).min(-1), P <0.01), particularly in the chylomicron fraction, so that absolute TG uptake was maintained despite lower plasma TG concentrations (control, 1.53 +/- 0.13 mmol/l; exercise, 1.01 +/- 0.16 mmol/l, P < 0.001). Prior exercise increased postprandial leg blood flow and glucose uptake (both P < 0.05). Mechanisms other than increased leg TG uptake must account for the effect of prior exercise on postprandial lipemia. PMID- 11052957 TI - Impaired myocardial protein synthesis induced by acute alcohol intoxication is associated with changes in eIF4F. AB - The purpose of the present study was to examine potential mechanisms for the known inhibitory effect of acute alcohol exposure on myocardial protein synthesis. Rats were injected intraperitoneally with either ethanol (75 mmol/kg) or saline, and protein synthesis was measured in vivo 2.5 h thereafter by use of the flooding-dose L-[(3)H]phenylalanine technique. Rates of myocardial protein synthesis and translational efficiency in alcohol-treated rats were decreased compared with control values. Free (nonpolysome bound) 40S and 60S ribosomal subunits were increased 50% after alcohol treatment, indicating an impaired peptide-chain initiation. To identify mechanisms responsible for this impairment, several eukaryotic initiation factors (eIF) were analyzed. Acute alcohol intoxication did not significantly alter the myocardial content of eIF2 alpha or eIF2B epsilon, the extent of eIF2 alpha phosphorylation, or the activity of eIF2B. Acute alcohol exposure increased the binding of 4E-binding protein 1 (4E BP1) to eIF4E (55%), diminished the amount of eIF4E bound to eIF4G (70%), reduced the amount of 4E-BP1 in the phosphorylated gamma-form (40%), and decreased the phosphorylation of p70S6 kinase and the ribosomal protein S6. There was no significant difference in either the plasma insulin-like growth factor (IGF) I concentration (total or free) or expression of IGF-I or IGF-II mRNA in heart between the two groups. These data suggest that the acute alcohol-induced impairment in myocardial protein synthesis results, in part, from an inhibition in peptide-chain initiation, which is associated with marked changes in eIF4E availability and p70S6 kinase phosphorylation but is independent of changes in the eIF2/2B system and IGFs. PMID- 11052958 TI - Lipid oxidation is reduced in obese human skeletal muscle. AB - The purpose of this study was to discern cellular mechanisms that contribute to the suppression of lipid oxidation in the skeletal muscle of obese individuals. Muscle was obtained from obese [body mass index (BMI), 38.3 +/- 3.1 kg/m(2)] and lean (BMI, 23.8 +/- 0.9 kg/m(2)) women, and fatty acid oxidation was studied by measuring (14)CO(2) production from (14)C-labeled fatty acids. Palmitate oxidation, which is at least partially dependent on carnitine palmitoyltransferase-1 (CPT-1) activity, was depressed (P < 0.05) by approximately 50% with obesity (6.8 +/- 2.2 vs. 13.7 +/- 1.4 nmole CO(2).g(-1).h( 1)). The CPT-1-independent event of palmitoyl carnitine oxidation was also depressed (P < 0.01) by approximately 45%. There were significant negative relationships (P < 0.05) for adiposity with palmitate (r = -0.76) and palmitoyl carnitine (r = -0.82) oxidation. Muscle CPT-1 and citrate synthase activity, an index of mitochondrial content, were also significantly (P < 0.05) reduced ( approximately 35%) with obesity. CPT-1 (r = -0.48) and citrate synthase (r = 0.65) activities were significantly (P < 0.05) related to adiposity. These data suggest that lesions at CPT-1 and post-CPT-1 events, such as mitochondrial content, contribute to the reduced reliance on fat oxidation evident in human skeletal muscle with obesity. PMID- 11052959 TI - Retinoic acid and host-pathogen interactions: effects on inducible nitric oxide synthase in vivo. AB - Vitamin A and its metabolite retinoic acid modulate the host response to pathogens through poorly characterized mechanisms. In vitro studies have suggested that retinoic acid decreases inducible NO synthase (NOS2, or iNOS) expression, a component of innate immunity, in several cell types stimulated with lipopolysaccharide (LPS) or cytokines. This study investigated the effect of retinoic acid on LPS-stimulated NOS2 expression in vivo. Wistar-Kyoto rats received all-trans retinoic acid (RA, 10 mg/kg) or vehicle intraperitoneally daily for 5 days followed by LPS (4 mg/kg) or saline intraperitoneally and were killed 6 h later. NOS2 activation was estimated by mRNA (RT-PCR) and protein (Western-blot) expression and plasma nitrate/nitrite accumulation. In sharp contrast to previous in vitro study reports, RA significantly enhanced NOS2 mRNA, protein expression, and plasma nitrate/nitrite concentration in LPS-injected rats but not in saline-injected rats. This was associated with increased expression of interleukin-2, interferon (IFN)-gamma and IFN regulatory factor-1 mRNAs in several organs and increased IFN-gamma plasma concentration. RA significantly increased mortality in LPS-injected rats. The NOS inhibitor aminoguanidine (50 mg/kg before LPS injection) significantly attenuated the RA-mediated increase in mortality. These results demonstrate for the first time that RA supplementation in vivo enhances activation of the LPS-triggered NOS2 pathway. PMID- 11052960 TI - Differential gender responses to hypoglycemia are due to alterations in CNS drive and not glycemic thresholds. AB - The aims of this study were 1) to determine whether differential glycemic thresholds are the mechanism responsible for the sexual dimorphism present in neuroendocrine responses during hypoglycemia and 2) to define the differences in counterregulatory physiological responses that occur over a range of mild to moderate hypoglycemia in healthy men and women. Fifteen (8 male, 7 female) lean healthy adults underwent four separate randomized 2-h hyperinsulinemic (1.5 mU. kg(-1).min(-1)) glucose clamp studies at euglycemia (90 mg/dl) or hypoglycemia of 70, 60, or 50 mg/dl. Plasma insulin levels were similar during euglycemic and hypoglycemic studies (91-96 +/- 8 microU/ml) in men and women. Hypoglycemia of 70, 60, and 50 mg/dl all resulted in significant increases (P < 0.05, P < 0.01) in epinephrine, glucagon, growth hormone, cortisol, and pancreatic polypeptide levels compared with euglycemic studies in men and women. Plasma norepinephrine levels were increased (P < 0.05) only relative to euglycemic studies at a hypoglycemia of 50 mg/dl. Muscle sympathetic nerve activity (MSNA) increased significantly during hyperinsulinemic-euglycemic control studies. Further elevations of MSNA did not occur until hypoglycemia of 60 mg/dl in both men and women. Plasma epinephrine, glucagon, growth hormone, and pancreatic polypeptide were significantly increased in men compared with women during hypoglycemia of 70, 60, and 50 mg/dl. MSNA, heart rate, and systolic blood pressure responses were also significantly increased in men at hypoglycemia of 60 and 50 mg/dl. In summary, these studies have demonstrated that, in healthy men and women, the glycemic thresholds for activation of epinephrine, glucagon, growth hormone, cortisol, and pancreatic polypeptide occur between 70 and 79 mg/dl. Thresholds for activation of MSNA occur between 60 and 69 mg/dl, whereas norepinephrine is not activated until glycemia is between 50 and 59 mg/dl. We conclude that 1) differential glycemic thresholds are not the cause of the sexual dimorphism present in counterregulatory responses to hypoglycemia; 2) reduced central nervous system efferent input appears to be the mechanism responsible for lowered neuroendocrine responses to hypoglycemia in women; and 3) physiological counterregulatory responses (neuroendocrine, cardiovascular, and autonomic nervous system) are reduced across a broad range of hypoglycemia in healthy women compared with healthy men. PMID- 11052961 TI - Limitations to basal and insulin-stimulated skeletal muscle glucose uptake in the high-fat-fed rat. AB - Rats fed a high-fat diet display blunted insulin-stimulated skeletal muscle glucose uptake. It is not clear whether this is due solely to a defect in glucose transport, or if glucose delivery and phosphorylation are also impaired. To determine this, rats were fed standard chow (control rats) or a high-fat diet (HF rats) for 4 wk. Experiments were then performed on conscious rats under basal conditions or during hyperinsulinemic euglycemic clamps. Rats received primed constant infusions of 3-O-methyl-[(3)H]glucose (3-O-MG) and [1-(14)C]mannitol. Total muscle glucose concentration and the steady-state ratio of intracellular to extracellular 3-O-MG concentration [which distributes based on the transsarcolemmal glucose gradient (TSGG)] were used to calculate glucose concentrations at the inner and outer sarcolemmal surfaces ([G](im) and [G](om), respectively) in soleus. Total muscle glucose was also measured in two fast twitch muscles. Muscle glucose uptake was markedly decreased in HF rats. In control rats, hyperinsulinemia resulted in a decrease in soleus TSGG compared with basal, due to increased [G](im). In HF rats during hyperinsulinemia, [G](im) also exceeded zero. Hyperinsulinemia also decreased muscle glucose in HF rats, implicating impaired glucose delivery. In conclusion, defects in extracellular and intracellular components of muscle glucose uptake are of major functional significance in this model of insulin resistance. PMID- 11052962 TI - Fatty acid transport protein-1 mRNA expression in skeletal muscle and in adipose tissue in humans. AB - Fatty acid transporter protein (FATP)-1 mRNA expression was investigated in skeletal muscle and in subcutaneous abdominal adipose tissue of 17 healthy lean, 13 nondiabetic obese, and 16 obese type 2 diabetic subjects. In muscle, FATP-1 mRNA levels were higher in lean women than in lean men (2.2 +/- 0.1 vs. 0.6 +/- 0.2 amol/microg total RNA, P < 0.01). FATP-1 mRNA expression was decreased in skeletal muscle in obese women both in nondiabetic and in type 2 diabetic patients (P < 0.02 vs. lean women in both groups), and in all women there was a negative correlation with basal FATP-1 mRNA level and body mass index (r = -0.74, P < 0.02). In men, FATP-1 mRNA was expressed at similar levels in the three groups both in skeletal muscle (0.6 +/- 0.2, 0.6 +/- 0.2, and 0.8 +/- 0.2 amol/microg total RNA in lean, obese, and type 2 diabetic male subjects) and in adipose tissue (0.9 +/- 0.2 amol/microg total RNA in the 3 groups). Insulin infusion (3 h) reduced FATP-1 mRNA levels in muscle in lean women but not in lean men. Insulin did not affect FATP-1 mRNA expression in skeletal muscle in obese nondiabetic or in type 2 diabetic subjects nor in subcutaneous adipose tissue in any of the three groups. These data show a gender-related difference in the expression of the fatty acid transporter FATP-1 in skeletal muscle of lean individuals and suggest that changes in FATP-1 expression may not contribute to a large extent to the alterations in fatty acid uptake in obesity and/or type 2 diabetes. PMID- 11052963 TI - Feeding stimulates protein synthesis in muscle and liver of neonatal pigs through an mTOR-dependent process. AB - Protein synthesis is repressed in both skeletal muscle and liver after a short term fast and is rapidly stimulated in response to feeding. Previous studies in rats and pigs have shown that the feeding-induced stimulation of protein synthesis is associated with activation of the 70-kDa ribosomal protein S6 kinase (S6K1) as well as enhanced binding of eukaryotic initiation factor eIF4E to eIF4G to form the active eIF4F complex. In cells in culture, hormones and nutrients regulate both of these events through a protein kinase termed the mammalian target of rapamycin (mTOR). In the present study, the involvement of mTOR in the feeding-induced stimulation of protein synthesis in skeletal muscle and liver was examined. Pigs at 7 days of age were fasted for 18 h, and then one-half of the animals were fed. In addition, one-half of the animals in each group were administered rapamycin (0.75 mg/kg) 2 h before feeding. The results reveal that treating 18-h fasted pigs with rapamycin, a specific inhibitor of mTOR, before feeding prevented the activation of S6K1 and the changes in eIF4F complex formation observed in skeletal muscle and liver after feeding. Rapamycin also ablated the feeding-induced stimulation of protein synthesis in liver. In contrast, in skeletal muscle, rapamycin attenuated, but did not prevent, the stimulation of protein synthesis in response to feeding. The results suggest that feeding stimulates hepatic protein synthesis through an mTOR-dependent process involving enhanced eIF4F complex formation and activation of S6K1. However, in skeletal muscle, these two processes may account for only part of the stimulation of protein synthesis, and thus additional steps may be involved in the response. PMID- 11052964 TI - Leptin responses to glucose infusions in obesity-prone rats. AB - The secretion of leptin is dually regulated. In fasting animals, plasma leptin concentrations reflect body fat stores, whereas the incremental leptin response to fasting or refeeding most likely reflects insulin-mediated energy flux and metabolism within adipocytes. Impaired secretion of leptin in either pathway could result in obesity. We therefore measured plasma leptin concentrations in fasted animals and plasma leptin concentrations after an intravenous glucose infusion in a rat model of obesity. Young Sprague-Dawley (S-D) and Fischer 344 (F344) rats had similar percent body fat and fasting glucose and fasting leptin concentrations. However, F344 animals had higher insulin concentrations and leptin responses to intravenous glucose than did the S-D animals. The animals were then fed a control or high-fat diet for 6 wk. High-fat fed animals gained more weight and body fat than did the control fed animals. Control and high-fat fed F344 animals gained approximately 40% (P < 0.0001) more weight and >100% (P < 0.01) more body fat than did the S-D animals. Fasting leptin concentrations and leptin concentrations after intravenous glucose infusions and feeding were more than double (P < 0.05) in F344 animals compared with S-D animals. Whether an animal is fed a control or high-fat diet had little effect on the leptin response to intravenous glucose. In conclusion, young, lean F344 animals, before the onset of obesity, demonstrated a greater acute leptin response to intravenous glucose than similarly lean S-D animals. After a 6-wk diet, F344 animals had a greater percent increase in body weight and insulin resistance and exhibited higher fasting leptin concentrations and a greater absolute leptin response to intravenous glucose compared with the S-D animals. The chronic diet (control or high fat) had little impact on the acute leptin response to intravenous glucose. F344 animals exhibit leptin resistance in young, lean animals and after aging and fat accumulation. PMID- 11052965 TI - Estimations of muscle interstitial insulin, glucose, and lactate in type 2 diabetic subjects. AB - Previous measurement of insulin in human muscle has shown that interstitial muscle insulin and glucose concentrations are approximately 30-50% lower than in plasma during hyperinsulinemia in normal subjects. The aims of this study were to measure interstitial muscle insulin and glucose in patients with type 2 diabetes to evaluate whether transcapillary transport is part of the peripheral insulin resistance. Ten patients with type 2 diabetes and ten healthy controls matched for sex, age, and body mass index were investigated. Plasma and interstitial insulin, glucose, and lactate (measured by intramuscular in situ-calibrated microdialysis) in the medial quadriceps femoris muscle were analyzed during a hyperinsulinemic euglycemic clamp. Blood flow in the contralateral calf was measured by vein plethysmography. At steady-state clamping, at 60-120 min, the interstitial insulin concentration was significantly lower than arterial insulin in both groups (409 +/- 86 vs. 1,071 +/- 99 pmol/l, P < 0.05, in controls and 584 +/- 165 vs. 1, 253 +/- 82 pmol/l, P < 0.05, in diabetic subjects, respectively). Interstitial insulin concentrations did not differ significantly between diabetic subjects and controls. Leg blood flow was significantly higher in controls (8.1 +/- 1.2 vs. 4.4 +/- 0.7 ml. 100 g(-1).min(-1) in diabetics, P < 0.05). Calculated glucose uptake was less in diabetic patients compared with controls (7.0 +/- 1.2 vs. 10.8 +/- 1.2 micromol. 100 g(-1).min(-1), P < 0.05, respectively). Arterial and interstitial lactate concentrations were both higher in the control group (1.7 +/- 0.1 vs. 1.2 +/- 0.1, P < 0. 01, and 1.8 +/- 0.1 vs. 1.2 +/- 0.2 mmol/l, P < 0.05, in controls and diabetics, respectively). We conclude that, during hyperinsulinemia, muscle interstitial insulin and glucose concentrations did not differ between patients with type 2 diabetes and healthy controls despite a significantly lower leg blood flow in diabetic subjects. It is suggested that decreased glucose uptake in type 2 diabetes is caused by insulin resistance at the cellular level rather than by a deficient access of insulin and glucose surrounding the muscle cell. PMID- 11052966 TI - Altered metabolism causes cardiac dysfunction in perfused hearts from diabetic (db/db) mice. AB - Contractile function and substrate metabolism were characterized in perfused hearts from genetically diabetic C57BL/KsJ-lepr(db)/lepr(db) (db/db) mice and their non-diabetic lean littermates. Contractility was assessed in working hearts by measuring left ventricular pressures and cardiac power. Rates of glycolysis, glucose oxidation, and fatty acid oxidation were measured using radiolabeled substrates ([5-(3)H]glucose, [U-(14)C]glucose, and [9,10-(3)H]palmitate) in the perfusate. Contractile dysfunction in db/db hearts was evident, with increased left ventricular end diastolic pressure and decreased left ventricular developed pressure, cardiac output, and cardiac power. The rate of glycolysis from exogenous glucose in diabetic hearts was 48% of control, whereas glucose oxidation was depressed to only 16% of control. In contrast, palmitate oxidation was increased twofold in db/db hearts. The hypothesis that altered metabolism plays a causative role in diabetes-induced contractile dysfunction was tested using perfused hearts from transgenic db/db mice that overexpress GLUT-4 glucose transporters. Both glucose metabolism and palmitate metabolism were normalized in hearts from db/db-human insulin-regulatable glucose transporter (hGLUT-4) hearts, as was contractile function. These findings strongly support a causative role of impaired metabolism in the cardiomyopathy observed in db/db diabetic hearts. PMID- 11052967 TI - Skeletal muscle apoptosis after burns is associated with activation of proapoptotic signals. AB - Critical illness is associated with muscle wasting and muscle weakness. Using burn injury as a model of local and systemic inflammatory response, we tested the hypothesis that thermal injury causes apoptosis in muscle. After a 40% body surface area burn to rats, abdominal muscles beneath the burn and limb muscles distant from the burn were examined for apoptosis at varying times after burn. Ladder assay, ELISA, and histological methods showed evidence of apoptosis in the abdominal muscles within 4-12 h with peak changes occurring at 3-7 days. Maximal apoptosis was also evident at distant limb muscles at 3-7 days. Investigation of proapoptotic pathways indicated mitochondrial membrane potential to be altered by 1 h after burn. Starting at 15 min after burn, cytochrome c was released from the mitochondria into the cytosol, followed by increased activity of caspase-3, starting at 6 h after burn. These studies suggest that mitochondria and caspase mediated apoptotic pathways may be an additional mechanism of muscle weight loss in burns and may be potential therapeutic targets for prevention of muscle wasting. PMID- 11052968 TI - Lumped constant for [(18)F]fluorodeoxyglucose in skeletal muscles of obese and nonobese humans. AB - Quantitative 2-[(18)F]fluoro-2-deoxy-D-glucose ([(18)F]FDG) positron emission tomography (PET) has been widely used to calculate glucose utilization in skeletal muscle. FDG-PET results depend partly on the lumped constant (LC), which accounts for the differences in the transport and phosphorylation between [(18)F]FDG and glucose. In this study, we estimated the LC for [(18)F]FDG directly in normal and in insulin-resistant obese subjects by combining FDG PET with the microdialysis technique. Eight obese [age 29.4 +/- 1.0 yr, body mass index (BMI) 33.6 +/- 1.0 kg/m(2)] and eight nonobese (age 25.0 +/- 1.0 yr, BMI 23.1 +/- 1.0 kg/m(2)) males were studied during euglycemic hyperinsulinemia (1 mU. kg(-1).min(-1) for 150 min). Muscle blood flow was measured using (15)O labeled water and PET. Muscle [(18)F]FDG uptake (rGU(FDG)) was calculated with Patlak graphic analysis. Interstitial glucose concentration of the quadriceps femoris muscle was measured simultaneously with [(18)F]FDG scanning using microdialysis. Muscle glucose uptake (by microdialysis, rGU(MD)) was calculated by multiplying glucose extraction by regional muscle blood flow. A significant correlation was found between rGU(MD) and rGU(FDG) (r = 0.78, P < 0.01). The LC was determined as the ratio of the rGU(FDG) to the rGU(MD). The LC averaged 1.16 +/- 0.16 and was similar in the obese and nonobese subjects (1.15 +/- 0.11 vs. 1.16 +/- 0.07, respectively, not significant). In conclusion, the microdialysis technique can be reliably combined with FDG PET to measure glucose uptake in skeletal muscle. Direct measurements with these two independent techniques suggest an LC value of 1.2 for [(18)F]FDG in human skeletal muscle during insulin stimulation, and the LC appears not to be sensitive to insulin resistance. PMID- 11052969 TI - Isoform-specific regulation of the lactate transporters MCT1 and MCT4 by contractile activity. AB - We examined the isoform-specific regulation of monocarboxylate transporter (MCT)1 and MCT4 expression by contractile activity in red and white tibialis anterior muscles. After 1 and 3 wk of chronic muscle stimulation (24 h/day), MCT1 protein expression was increased in the red muscles (+78%, P < 0.05). In the white muscles, MCT1 was increased after 1 wk (+191%) and then was decreased after 3 wk. In the red muscle, MCT1 mRNA accumulation was increased only after 3 wk (+21%; P < 0.05). In the white muscle, MCT1 mRNA was increased after 1 wk (+30%; P < 0.05) and 3 wk (+15%; P < 0.05). MCT4 protein was not altered in either the red or white muscles after 1 or 3 wk. MCT4 mRNA was transiently lowered (approximately 15%) in both muscles in the 1st wk, but MCT4 mRNA levels were back to control levels after 3 wk. In conclusion, chronic contractile activity induces the expression of MCT1 but not MCT4. This increase in MCT1 alone was sufficient to increase lactate uptake from the circulation. PMID- 11052970 TI - Age-induced changes in pancreatic islet blood flow: evidence for an impaired regulation in diabetic GK rats. AB - The present study aimed to compare longitudinal variations in islet blood perfusion in rats with different degrees of impairment of glucose metabolism. For this purpose, mildly diabetic Goto-Kakizaki (GK) rats, glucose intolerant F(1) hybrids of GK and Wistar (W) rats (H), and control W rats were examined at 5 wk, 12 wk, or 1 yr of age, using the microsphere technique for blood flow measurements. W rats showed progressively increasing islet blood flow (IBF) throughout the experiment. Both GK and H rats demonstrated increasing IBF between 5 and 12 wk. However, H rats showed no further increment in IBF at 1 yr, whereas GK rats displayed a pronounced decrease in IBF between 12 wk and 1 yr of age. The augmented IBF seen in older W rats may constitute an adaptation to the increasing demand for insulin secretion in aging rats. The inability to adapt to the increased demand for insulin secretion by upregulation of islet blood flow could contribute to the progressive deterioration of glucose metabolism seen in the aging GK rat. PMID- 11052971 TI - IGF-I/IGFBP-3 binary complex modulates sepsis-induced inhibition of protein synthesis in skeletal muscle. AB - The present study evaluated the ability of insulin-like growth factor I (IGF-I) complexed with IGF binding protein-3 (IGFBP-3) to modulate the sepsis-induced inhibition of protein synthesis in gastrocnemius. Beginning 16 h after the induction of sepsis, either the binary complex or saline was injected twice daily via a tail vein, with measurements made 3 and 5 days later. By day 3, sepsis had reduced plasma IGF-I concentrations approximately 50% in saline-treated rats. Administration of the binary complex provided exogenous IGF-I to compensate for the sepsis-induced diminished plasma IGF-I. Sepsis decreased rates of protein synthesis in gastrocnemius relative to controls by limiting translational efficiency. Treatment of septic rats with the binary complex for 5 days attenuated the sepsis-induced inhibition of protein synthesis and restored translational efficiency to control values. Assessment of potential mechanisms regulating translational efficiency showed that neither the sepsis-induced change in gastrocnemius content of eukaryotic initiation factor 2B (eIF2B), the amount of eIF4E associated with 4E binding protein-1 (4E-BP1), nor the phosphorylation state of 4E-BP1 or eIF4E were altered by the binary complex. Overall, the results are consistent with the hypothesis that decreases in plasma IGF-I are partially responsible for enhanced muscle catabolism during sepsis. PMID- 11052972 TI - Effects of high-dose estrogen on murine hematopoietic bone marrow precede those on osteogenesis. AB - High-dose estrogen both stimulates new medullary bone formation and suppresses hematopoiesis in mouse long bones. To determine whether the latter response is a direct consequence of the former, we compared the time course of estrogen's effects on osteogenesis and hematopoietic bone marrow. Flow cytometry was employed to measure hematopoietic subpopulations in bone marrow from femurs of female mice killed at different times after commencing 0.5 mg estradiol/wk to each animal. Estrogen markedly reduced the number of leucocytes (CD11a positive), which had already diminished by 75% after 4 days and had virtually disappeared by 18 days. Specific populations showed a similar pattern of decline after estrogen, including B lymphocytes, monocytes, and endothelial cells. In contrast, the osteogenic precursor population showed a marked increase after estrogen treatment, as assessed by assaying alkaline phosphatase-positive colony-forming units (fibroblastic) ex vivo. However, this rise did not reach significance until 8 days after estrogen administration, suggesting that it follows rather than precedes estrogen's effects on hematopoiesis. We conclude that estrogen does not suppress hematopoiesis in mouse long bones as a direct consequence of its effects on osteogenesis. PMID- 11052974 TI - Effect of sepsis on eIE4E availability in skeletal muscle. AB - Chronic septic abscess formation causes an inhibition of protein synthesis in gastrocnemius that is not observed in rats with a sterile abscess. The inhibition is associated with an impaired translation initiation. The present study was designed to investigate the effects of sepsis on phosphorylation and availability of eukaryotic initiation factor (eIF)4E in gastrocnemius 5 days after induction of a sterile or septic abscess. Neither sepsis nor sterile inflammation altered the extent of eIF4E phosphorylation. Moreover, no changes in the amount of the binding protein 4E-BP1 associated with eIF4E or in the phosphorylation of 4E-BP1 were observed during sepsis or sterile inflammation. In contrast, sepsis and sterile inflammation caused a reduction in the relative amount of eIF4G bound to eIF4E compared with controls. The diminished amount of eIF4G bound to eIF4E was not the result of a reduced abundance of eIF4E. Sepsis, but not sterile inflammation, caused an increase in the cellular abundance of eIF4E. The results provide evidence that alterations in the eIF4E system are probably not rate controlling for the synthesis of total, mixed proteins in gastrocnemius during sepsis. Instead, on the basis of our previous studies, changes in eIF2B appear to be responsible for limiting protein synthesis in skeletal muscle during sepsis. PMID- 11052973 TI - Alterations in basal glucose metabolism during late pregnancy in the conscious dog. AB - We assessed basal glucose metabolism in 16 female nonpregnant (NP) and 16 late pregnant (P) conscious, 18-h-fasted dogs that had catheters inserted into the hepatic and portal veins and femoral artery approximately 17 days before the experiment. Pregnancy resulted in lower arterial plasma insulin (11 +/- 1 and 4 +/- 1 microU/ml in NP and P, respectively, P < 0.05), but plasma glucose (5.9 +/- 0.1 and 5.6 +/- 0.1 mg/dl in NP and P, respectively) and glucagon (39 +/- 3 and 36 +/- 2 pg/ml in NP and P, respectively) were not different. Net hepatic glucose output was greater in pregnancy (42.1 +/- 3.1 and 56.7 +/- 4.0 micromol. 100 g liver(-1).min(-1) in NP and P, respectively, P < 0.05). Total net hepatic gluconeogenic substrate uptake (lactate, alanine, glycerol, and amino acids), a close estimate of the gluconeogenic rate, was not different between the groups (20.6 +/- 2.8 and 21.2 +/- 1.8 micromol. 100 g liver(-1). min(-1) in NP and P, respectively), indicating that the increment in net hepatic glucose output resulted from an increase in the contribution of glycogenolytically derived glucose. However, total glycogenolysis was not altered in pregnancy. Ketogenesis was enhanced nearly threefold by pregnancy (6.9 +/- 1.2 and 18.2 +/- 3.4 micromol. 100 g liver(-1).min(-1) in NP and P, respectively), despite equivalent net hepatic nonesterified fatty acid uptake. Thus late pregnancy in the dog is not accompanied by changes in the absolute rates of gluconeogenesis or glycogenolysis. Rather, repartitioning of the glucose released from glycogen is responsible for the increase in hepatic glucose production. PMID- 11052975 TI - Glucocorticoids oppose translational control by leucine in skeletal muscle. AB - Glucocorticoids comprise an important class of hormonal mediators of fuel and protein homeostasis in normal and pathological scenarios. In skeletal muscle, exposure to glucocorticoids is characterized by a reduction in protein synthetic rate coincident with hampered translation initiation. However, it is unclear whether this involves attenuation of anabolic stimuli or is simply due to inhibition of the basally activated translational apparatus. Therefore, this inquiry was designed to determine whether leucine, administered orally, could rescue the translational inhibition induced by glucocorticoids. Dexamethasone, injected intraperitoneally, acutely diminished protein synthetic rates to 80% of control values in skeletal muscle from rat hindlimb. The eukaryotic initiation factor (eIF)4 regulatory element was simultaneously and negatively impacted via sequestration of eIF4E by the hypophosphorylated form of the translational suppressor, eIF4E binding protein 1 (4E-BP1). The 70-kDa ribosomal protein S6 kinase (S6K1) was also dephosphorylated, notably at T389, in response to glucocorticoids. Leucine, administered orally, effectively restored each aforementioned translational parameter to control levels. Inasmuch as leucine's potency in modulation of the translational machinery, and indeed of protein turnover in general, is widely appreciated, this amino acid may prove useful in normalizing the impairment of mRNA translation associated with various muscle wasting pathologies, such as glucocorticoid excess. PMID- 11052976 TI - Induction of GLUT-1 protein in adult human skeletal muscle fibers. AB - Prompted by our recent observations that GLUT-1 is expressed in fetal muscles, but not in adult muscle fibers, we decided to investigate whether GLUT-1 expression could be reactivated. We studied different stimuli concerning their ability to induce GLUT-1 expression in mature human skeletal muscle fibers. Metabolic stress (obesity, non-insulin-dependent diabetes mellitus), contractile activity (training), and conditions of de- and reinnervation (amyotrophic lateral sclerosis) could not induce GLUT-1 expression in human muscle fibers. However, regenerating muscle fibers in polymyositis expressed GLUT-1. In contrast to GLUT 1, GLUT-4 was expressed in all investigated muscle fibers. Although the significance of GLUT-1 in adult human muscle fibers appears limited, GLUT-1 may be of importance for the glucose supplies in immature and regenerating muscle. PMID- 11052977 TI - Acute reversal of lipid-induced muscle insulin resistance is associated with rapid alteration in PKC-theta localization. AB - Muscle insulin resistance in the chronic high-fat-fed rat is associated with increased membrane translocation and activation of the novel, lipid-responsive, protein kinase C (nPKC) isozymes PKC-theta and -epsilon. Surprisingly, fat induced insulin resistance can be readily reversed by one high-glucose low-fat meal, but the underlying mechanism is unclear. Here, we have used this model to determine whether changes in the translocation of PKC-theta and -epsilon are associated with the acute reversal of insulin resistance. We measured cytosol and particulate PKC-alpha and nPKC-theta and -epsilon in muscle in control chow-fed Wistar rats (C) and 3-wk high-fat-fed rats with (HF-G) or without (HF-F) a single high-glucose meal. PKC-theta and -epsilon were translocated to the membrane in muscle of insulin-resistant HF-F rats. However, only membrane PKC-theta was reduced to the level of chow-fed controls when insulin resistance was reversed in HF-G rats [% PKC-theta at membrane, 23.0 +/- 4.4% (C); 39.7 +/- 3.4% (HF-F, P < 0.01 vs. C); 22.5 +/- 2.7% (HF-G, P < 0.01 vs. HF-F), by ANOVA]. We conclude that, although muscle localization of both PKC-epsilon and PKC-theta are influenced by chronic dietary lipid oversupply, PKC-epsilon and PKC-theta localization are differentially influenced by acute withdrawal of dietary lipid. These results provide further support for an association between PKC-theta muscle cellular localization and lipid-induced muscle insulin resistance and stress the labile nature of high-fat diet-induced insulin resistance in the rat. PMID- 11052978 TI - AMPK signaling in contracting human skeletal muscle: acetyl-CoA carboxylase and NO synthase phosphorylation. AB - AMP-activated protein kinase (AMPK) is a metabolic stress-sensing protein kinase responsible for coordinating metabolism and energy demand. In rodents, exercise accelerates fatty acid metabolism, enhances glucose uptake, and stimulates nitric oxide (NO) production in skeletal muscle. AMPK phosphorylates and inhibits acetyl coenzyme A (CoA) carboxylase (ACC) and enhances GLUT-4 translocation. It has been reported that human skeletal muscle malonyl-CoA levels do not change in response to exercise, suggesting that other mechanisms besides inhibition of ACC may be operating to accelerate fatty acid oxidation. Here, we show that a 30-s bicycle sprint exercise increases the activity of the human skeletal muscle AMPK-alpha1 and -alpha2 isoforms approximately two- to threefold and the phosphorylation of ACC at Ser(79) (AMPK phosphorylation site) approximately 8.5-fold. Under these conditions, there is also an approximately 5.5-fold increase in phosphorylation of neuronal NO synthase-mu (nNOSmu;) at Ser(1451). These observations support the concept that inhibition of ACC is an important component in stimulating fatty acid oxidation in response to exercise and that there is coordinated regulation of nNOSmu to protect the muscle from ischemia/metabolic stress. PMID- 11052979 TI - Fibrogenesis. V. TGF-beta signaling pathways. AB - Transforming growth factor (TGF)-beta is a multifunctional peptide growth factor with a wide range of potential effects on growth, differentiation, extracellular matrix deposition, and the immune response. General TGF-beta signaling pathways have been described in detail over the last several years, but factors that determine the nature of the TGF-beta response are poorly understood. In particular, signaling pathways that specifically mediate the matrix effects of TGF-beta have received little attention, although they will be important therapeutic targets in the treatment of pathological fibrosis. This themes article focuses on TGF-beta signaling and highlights potential points for generating matrix-specific responses. PMID- 11052980 TI - Molecular physiology and pathophysiology of tight junctions. IV. Regulation of tight junctions by extracellular stimuli: nutrients, cytokines, and immune cells. AB - The epithelial lining of the gastrointestinal tract forms a regulated, selectively permeable barrier between luminal contents and the underlying tissue compartments. Permeability across the epithelium is, in part, determined by the rate-limiting barrier of the paracellular pathway-the most apical intercellular junction referred to as the tight junction (TJ). The TJ is composed of a multiprotein complex that affiliates with the underlying apical actomyosin ring. TJ structure and function, and therefore epithelial permeability, are influenced by diverse physiological and pathological stimuli; here we review examples of such stimuli that are detected at the cell surface. For example, luminal glucose induces an increase in paracellular permeability to small molecules. Similarly, but by other means, cytokines and leukocytes in the vicinity of the epithelium also regulate TJ structure and paracellular permeability by influencing the TJ protein complex and/or its association with the underlying actin cytoskeleton. PMID- 11052981 TI - Disruption of cyclooxygenase-1 gene results in an impaired response to radiation injury. AB - Prostaglandins may play an important role in regulating normal renewal of gastrointestinal epithelium, epithelial injury repair, and initiation or progression of intestinal neoplasia. Synthesis of prostaglandins is catalyzed by either of two cyclooxygenase isoforms, Cox-1 and Cox-2. Cox-1 is the predominant cyclooxygenase isoform found in the normal intestine. In contrast, Cox-2 is present at low levels in normal intestine but is elevated at sites of inflammation and in adenomas and carcinomas. To determine directly whether prostaglandins synthesized by Cox-1 or Cox-2 regulate crypt epithelial cell fate after genotoxic or cytotoxic injury, we examined apoptosis, prostaglandin synthesis, and crypt stem cell survival after gamma-irradiation in Cox-1(-/-) and Cox-2(-/-) mice. Cox-1(-/-) mice had increased crypt epithelial cell apoptosis and decreased clonogenic stem cell survival compared with wild-type littermates. PGE(2) synthesis was also diminished in Cox-1(-/-) mice compared with wild-type controls in unstressed intestine and after radiation injury. In contrast, apoptosis, stem cell survival, and intestinal PGE(2) synthesis in Cox-2(-/-) mice after irradiation were the same as in wild-type littermates. Crypt stem cell survival after irradiation was inhibited by a highly specific neutralizing antibody to PGE(2), suggesting that this prostaglandin mediates stem cell fate in vivo. These data suggest that prostaglandins synthesized by Cox-1 regulate multiple steps that determine the fate of crypt epithelial cell after genotoxic or cytotoxic injury. PMID- 11052982 TI - A primary culture of guinea pig gallbladder epithelial cells that is responsive to secretagogues. AB - We have developed a cell culture of guinea pig gallbladder epithelial cells with which to study ion transport. When grown on permeable supports, the cultured epithelia developed a transepithelial resistance (R(t)) of approximately 500 Omega. cm(2). The epithelial cell origin of the cell culture was further confirmed by immunocytochemical localization of cytokeratin. Ionomycin and forskolin increased transepithelial voltage and short-circuit current (I(sc)) and decreased R(t). The response to ionomycin was transient, whereas that to forskolin was sustained. Both were attenuated by replacement of Cl(-) and/or HCO(3)(-). Mucosal addition of the anion transport inhibitors DIDS or diphenylamine-2-carboxylic acid (DPC) blocked the response to ionomycin. The response to forskolin was blocked by DPC but not by DIDS. Ionomycin, but not forskolin, increased intracellular Ca(2+) concentration in fura 2-loaded cells. PGE(2), histamine, vasoactive intestinal polypeptide, and secretin elicited a sustained increase in I(sc). Responses to ATP and CCK were transient. Thus cultured guinea pig gallbladder epithelia display the range of responses observed in the native tissue and are an appropriate model for studies of ion transport in gallbladder and intestinal epithelia. PMID- 11052983 TI - Cytochalasin B modulation of Caco-2 tight junction barrier: role of myosin light chain kinase. AB - The intracellular mechanisms that mediate cytochalasin-induced increase in intestinal epithelial tight junction (TJ) permeability are unclear. In this study, we examined the involvement of myosin light chain kinase (MLCK) in this process, using the filter-grown Caco-2 intestinal epithelial monolayers. Cytochalasin B (Cyto B) (5 microg/ml) produced an increase in Caco-2 MLCK activity, which correlated with the increase in Caco-2 TJ permeability. The inhibition of Cyto B-induced MLCK activation prevented the increase in Caco-2 TJ permeability. Additionally, myosin-Mg(2+)-ATPase inhibitor and metabolic inhibitors (which inhibit MLCK induced actin-myosin contraction) also prevented the Cyto B-induced increase in Caco-2 TJ permeability. Cyto B caused a late-phase (15-30 min) aggregation of actin fragments into large actin clumps, which was also inhibited by MLCK inhibitors. Cyto B produced a morphological disturbance of the ZO-1 TJ proteins, visually correlating with the functional increase in Caco-2 TJ permeability. The MLCK and myosin-Mg(2+)-ATPase inhibitors prevented both the functional increase in TJ permeability and disruption of ZO-1 proteins. These findings suggested that Cyto B-induced increase in Caco-2 TJ permeability is regulated by MLCK activation. PMID- 11052984 TI - Portal hypertension induces sodium channel expression in colonocytes from the distal colon of the rat. AB - Cellular mechanisms for Na(+) retention in portal hypertension are undefined, but epithelial Na(+) channels (ENaC) may be involved. Under high-salt diet, ENaC are absent from distal colon of rat but can be induced by mineralocorticoids such as aldosterone. Presence of rat ENaC was determined by amiloride inhibition of (22)Na(+) uptake in surface colonocytes 7 and 14 days after partial portal vein ligation (PVL) or sham surgery. At both times, uptake inhibition was significantly increased in PVL rats. Presence of mRNA transcripts, determined by RT-PCR, demonstrated that channel alpha- and gamma-subunits were similarly expressed in both groups but that beta-subunit mRNA was increased in PVL rats. This confirms that there was induction of rat ENaC and indicates that beta subunit has a regulatory role. Urinary Na(+) was decreased for 3 days after PVL but was not different at other times, and serum aldosterone levels were elevated at 7 days, at a time when urinary Na(+) output was similar to that of sham operated rats. We conclude that PVL leads to induction of ENaC in rat distal colon. An increase in aldosterone levels may prevent natiuresis and is probably one of several control mechanisms involved in Na(+) retention in portal hypertension. PMID- 11052985 TI - Actions of reactive oxygen species on AH/type 2 myenteric neurons in guinea pig distal colon. AB - With conventional intracellular recording methods, we investigated the mechanism of actions of reactive oxygen species (ROS) derived from hypoxanthine and xanthine oxidase (HX/XO) reactions on AH/type 2 myenteric neurons in the guinea pig distal colon. Of the 54 neurons to which HX/XO was applied, 32 neurons showed a transient membrane hyperpolarization(s) followed by a long-lasting membrane depolarization. Two additional groups of 10 myenteric neurons exhibited only a membrane hyperpolarization(s) or a late-onset membrane depolarization, respectively, and the remaining two neurons did not show any response to HX/XO. Analysis of changes of the input resistance induced by HX/XO indicated that suppression and augmentation of the conductance of Ca(2+)-dependent K(+) channels are the ionic mechanisms underlying the membrane hyperpolarization and depolarization, respectively. The effects of HX/XO on myenteric neurons were mimicked by application of caffeine or H(2)O(2). The results suggest that OH(.), but neither H(2)O(2) nor O(2)(.-), is responsible for HX/XO-induced responses. The intracellular Ca(2+) store may be the acting site of ROS in colonic AH/type 2 neurons. PMID- 11052986 TI - Metabolism and acid secretory effect of sulfated and nonsulfated gastrin-6 in humans. AB - The antral hormone gastrin is synthesized by processing progastrin into different peptides that stimulate gastric secretion. The effect on acid secretion depends mainly on the metabolic clearance rate of the peptides, but some of them may differ in potency and maximum acid output at similar concentrations in plasma. Sulfated and nonsulfated gastrin-6 are the smallest circulating bioactive gastrins in humans. Their effect and metabolism have now been investigated in nine normal subjects and compared with nonsulfated gastrin-17, a main product of progastrin. Maximum acid output after stimulation with gastrin-17, sulfated gastrin-6, and nonsulfated gastrin-6 were 28.3 +/- 2.0, 24.5 +/- 2.0 (P < 0.02), and 19.3 +/- 2. 3 (P < 0.05) mmol H(+)/50 min, respectively, and the corresponding EC(50) values were 43 +/- 6, 24 +/- 2 (P < 0.01), and 25 +/- 2 (not significant) pmol/l. The half-life of gastrin-17 was 5.3 +/- 0.3 min, the metabolic clearance rate (MCR) was 16.5 +/- 1.3 ml. kg(-1). min(-1), and the apparent volume of distribution (V(d)) was 124.3 +/- 9.6 ml/kg. The half-lives of sulfated and nonsulfated gastrin-6 were 2.1 +/- 0.3 and 1.9 +/- 0.3 min, the MCRs were 42.8 +/- 3.7 and 139.4 +/- 9.6 ml kg(-1) min(-1) (P < 0.01), and the V(d) were 139.0 +/- 30.5 and 392.0 +/- 81.6 (P < 0.01) ml kg(-1). All pharmacokinetic parameters differed significantly from gastrin-17 (P < 0.01). We conclude that gastrin 6 has a higher potency but a lower efficacy than gastrin-17. The efficacy of gastrin-6 is increased by tyrosine O-sulfation, which also enhances the protection against elimination. PMID- 11052987 TI - Neural pathways regulating Brunner's gland secretion in guinea pig duodenum in vitro. AB - This study examined the neural pathways innervating Brunner's glands using a novel in vitro model of acinar secretion from Brunner's glands in submucosal preparations from the guinea pig duodenum. Neural pathways were activated by focal electrical stimulation and excitatory agonists, and videomicroscopy was used to monitor dilation of acinar lumen. Electrical stimulation of perivascular nerves evoked large dilations that were blocked by TTX (1 microM) or the muscarinic receptor antagonist 4-diphenylacetoxy-N-(2-chloroethyl)-piperidine hydrochloride (1 microM). The nicotinic agonist 1,1-dimethyl-4-phenylpiperazinium iodide (100 microM) had no effect, and the nerve-evoked responses were not inhibited by hexamethonium (200 microM). Dilations were abolished in preparations from chronically vagotomized animals. Activation of submucosal ganglia significantly dilated submucosal arterioles but not Brunner's glands. Effects of electrical stimulation of perivascular and submucosal nerves were not altered by guanethidine. Capsaicin and substance P also dilated arterioles but had no effect on Brunner's glands. Cholinergic (choline acetyltransferase-immunoreactive) nerve fibers were found in Brunner's glands. These findings demonstrate that Brunner's glands are innervated by cholinergic vagal fibers but not by capsaicin-sensitive or intrinsic enteric nerves. PMID- 11052988 TI - Butyrate upregulates stromelysin-1 production by intestinal mesenchymal cells. AB - Nutritional factors and resident bacteria participate in the pathogenesis of intestinal inflammation. However, the ways in which bacteria and complex diets might modulate matrix metalloproteinase (MMP) production are unknown. We hypothesized that butyrate might enhance production of MMPs, thus amplifying their response to signals in inflammatory conditions. Human mesenchymal cells were incubated with butyrate and then stimulated with cytokines. MMPs and inhibitors were studied by Western blotting and quantitative RT-PCR. Acetylation of histones was examined in Triton X acetic acid-urea gels by PAGE. We showed that butyrate selectively enhanced the protein production and mRNA expression of stromelysin-1 in tumor necrosis factor-alpha- or interleukin-1beta-stimulated mesenchymal cells. Butyrate alone did not induce any change in MMP production or mRNA expression. It increased the acetylation of histones in mesenchymal cells. Furthermore, acetylation of histones (induced by trichostatin A) reproduced the effects of butyrate. Although butyrate is a major source of nutrient for the colonic epithelial cells, it modulates intestinal inflammation through the secretion of stromelysin-1 in stimulated stromal cells via the inhibition of histone deacetylase. PMID- 11052989 TI - Ileal short-chain fatty acids inhibit gastric motility by a humoral pathway. AB - The aim of this study was to evaluate the nervous and humoral pathways involved in short-chain fatty acid (SCFA)-induced ileal brake in conscious pigs. The role of extrinsic ileal innervation was evaluated after SCFA infusion in innervated and denervated Babkin's ileal loops, and gastric motility was measured with strain gauges. Peptide YY (PYY) and glucagon-like peptide-1 (GLP-1) concentrations were evaluated in both situations. The possible involvement of absorbed SCFA was tested by using intravenous infusion of acetate. Ileal SCFA infusion in the intact terminal ileum decreased the amplitude of distal and terminal antral contractions (33 +/- 1.2 vs. 49 +/- 1.2% of the maximal amplitude recorded before infusion) and increased their frequency (1.5 +/- 0.11 vs. 1.3 +/- 0.10/min). Similar effects were observed during SCFA infusion in ileal innervated and denervated loops (amplitude, 35 +/- 1.0 and 34 +/- 0. 8 vs. 47 +/- 1.3 and 43 +/- 1.2%; frequency, 1.4 +/- 0.07 and 1.6 +/- 0.06 vs. 1.1 +/- 0.14 and 1.0 +/- 0.12/min). Intravenous acetate did not modify the amplitude and frequency of antral contractions. PYY but not GLP-1 concentrations were increased during SCFA infusion in innervated and denervated loops. In conclusion, ileal SCFA inhibit distal gastric motility by a humoral pathway involving the release of an inhibiting factor, which is likely PYY. PMID- 11052990 TI - Regulation of DRA and AE1 in rat colon by dietary Na depletion. AB - Two distinct Cl/anion exchange activities (Cl/HCO(3) and Cl/OH) identified in apical membranes of rat distal colon are distributed in cell type-specific patterns. Cl/HCO(3) exchange is expressed only in surface cells, whereas Cl/OH exchange is localized in surface and crypt cells. Dietary Na depletion substantially inhibits Cl/HCO(3) but not Cl/OH exchange. We determined whether anion exchange isoforms (AE) and/or downregulated in adenoma (DRA) are expressed in and related to apical membrane anion exchanges by examining localization of AE isoform-specific and DRA mRNA expression in normal and Na-depleted rats. Amplification of AE cDNA fragments by RT-PCR with colonic mRNA as template indicates that AE1 and AE2 but not AE3 mRNAs are expressed. In situ hybridization study revealed that AE1 mRNA is expressed predominantly in surface but not crypt cells. In contrast, AE2 polypeptide is expressed in basolateral membranes and DRA protein is expressed in apical membranes of both surface and crypt cells. AE1 mRNA is only minimally present in proximal colon, and DRA mRNA abundance is similar in distal and proximal colon. Dietary Na depletion reduces AE1 mRNA abundance but did not alter DRA mRNA abundance. This indicates that AE1 encodes surface cell-specific aldosterone-regulated Cl/HCO(3) exchange, whereas DRA encodes aldosterone-insensitive Cl/OH exchange. PMID- 11052991 TI - Activation of Na(+)- and Ca(2+)-dependent Mg(2+) extrusion by alpha(1)- and beta adrenergic agonists in rat liver cells. AB - The administration of selective alpha(1) (phenylephrine)-, beta (isoproterenol)-, or mixed (epinephrine) adrenergic agonists induces a marked Mg(2+) extrusion from perfused rat livers. In the absence of extracellular Ca(2+), phenylephrine does not induce a detectable Mg(2+) extrusion, isoproterenol-induced Mg(2+) mobilization is unaffected, and epinephrine induces a net Mg(2+) extrusion that is lower than in the presence of extracellular Ca(2+) and quantitatively similar to that elicited by isoproterenol. In the absence of extracellular Na(+), no Mg(2+) is extruded from the liver irrespective of the agonist used. Similar results are observed in perfused livers stimulated by glucagon or 8 chloroadenosine 3', 5'-cyclic monophosphate. In the absence of extracellular Na(+) or Ca(2+), adrenergic-induced glucose extrusion from the liver is also markedly decreased. Together, these results indicate that liver cells extrude Mg(2+) primarily via a Na(+)-dependent mechanism. This extrusion pathway can be activated by the increase in cellular cAMP that follows the stimulation by glucagon or a specific beta-adrenergic receptor agonist or, alternatively, by the changes in cellular Ca(2+) induced by the stimulation of the alpha(1) adrenoceptor. In addition, the stimulation of the alpha(1)-adrenoceptor appears to activate an auxiliary Ca(2+)-dependent Mg(2+) extrusion pathway. Finally, our data suggest that experimental conditions that affect Mg(2+) mobilization also interfere with glucose extrusion from liver cells. PMID- 11052992 TI - Inwardly rectifying K(+) channels in esophageal smooth muscle. AB - The whole cell patch-clamp technique was used to investigate whether there were inwardly rectifying K(+) (K(ir)) channels in the longitudinal muscle of cat esophagus. Inward currents were observable on membrane hyperpolarization negative to the K(+) equilibrium potential (E(k)) in freshly isolated esophageal longitudinal muscle cells. The current-voltage relationship exhibited strong inward rectification with a reversal potential (E(rev)) of -76.5 mV. Elevation of external K(+) increased the inward current amplitude and positively shifted its E(rev) after the E(k), suggesting that potassium ions carry this current. External Ba(2+) and Cs(+) inhibited this inward current, with hyperpolarization remarkably increasing the inhibition. The IC(50) for Ba(2+) and Cs(+) at -60 mV was 2.9 and 1.6 mM, respectively. Furthermore, external Ba(2+) of 10 microM moderately depolarized the resting membrane potential of the longitudinal muscle cells by 6.3 mV while inhibiting the inward rectification. We conclude that K(ir) channels are present in the longitudinal muscle of cat esophagus, where they contribute to its resting membrane potential. PMID- 11052993 TI - Retention of mutant alpha(1)-antitrypsin Z in endoplasmic reticulum is associated with an autophagic response. AB - Although there is evidence for specific subcellular morphological alterations in response to accumulation of misfolded proteins in the endoplasmic reticulum (ER), it is not clear whether these morphological changes are stereotypical or if they depend on the specific misfolded protein retained. This issue may be particularly important for mutant secretory protein alpha(1)-antitrypsin (alpha(1)AT) Z because retention of this mutant protein in the ER can cause severe target organ injury, the chronic hepatitis/hepatocellular carcinoma associated with alpha(1)AT deficiency. Here we examined the morphological changes that occur in human fibroblasts engineered for expression and ER retention of mutant alpha(1)ATZ and in human liver from three alpha(1)AT-deficient patients. In addition to marked expansion and dilatation of ER, there was an intense autophagic response. Mutant alpha(1)ATZ molecules were detected in autophagosomes by immune electron microscopy, and intracellular degradation of alpha(1)ATZ was partially reduced by chemical inhibitors of autophagy. In contrast to mutant CFTRDeltaF508, expression of mutant alpha(1)ATZ in heterologous cells did not result in the formation of aggresomes. These results show that ER retention of mutant alpha(1)ATZ is associated with a marked autophagic response and raise the possibility that autophagy represents a mechanism by which liver of alpha(1)AT-deficient patients attempts to protect itself from injury and carcinogenesis. PMID- 11052994 TI - Regulation of IGFBP-4 levels in human intestinal muscle by an IGF-I-activated, confluence-dependent protease. AB - Human intestinal smooth muscle cells in culture produce insulin-like growth factor-I (IGF-I), IGF binding protein-3 (IGFBP-3), IGFBP-4, and IGFBP-5, which can modulate the effects of IGF-I on growth. This study examined the role of IGFBP-4 on IGF-I-induced growth and the mechanisms regulating IGFBP-4 levels. IGFBP-4 inhibited IGF-I-induced [(3)H]thymidine incorporation. IGFBP-4 mRNA levels were not altered by IGF-I. IGF-I caused a concentration-dependent activation of an endogenous IGFBP-4 protease, resulting in time-dependent degradation of intact IGFBP-4 into inactive fragments. Protease activity was measured in a cell-free assay using smooth muscle cell conditioned medium containing the IGFBP-4 protease. The protease was inhibited by EDTA and benzamidine. Protease activity was highest in proliferating cells and lowest in postconfluent cells. The role of endogenous IGF-I in regulating IGFBP-4 degradation was confirmed by the ability of an IGF-I antagonist to inhibit IGF-I activated IGFBP-4 proteolysis in intact cells. We conclude that in human intestinal smooth muscle cells levels of secreted IGFBP-4 are determined by the confluence-dependent production of a cation-dependent serine protease that is activated by endogenous IGF-I. PMID- 11052995 TI - Somatostatin receptor subtype-5 mediates inhibition of peptide YY secretion from rat intestinal cultures. AB - Somatostatin-14 (S-14) and somatostatin-28 (S-28) bind to five distinct membrane receptors (SSTRs), but S-28 has higher affinity for SSTR-5. Whether S-28 acting through SSTR-5 regulates inhibition of peptide YY (PYY) secretion was tested in fetal rat intestinal cell cultures. S-28 and S-14 caused dose-dependent inhibition of PYY secretion stimulated by gastrin-releasing peptide, but S-28 was more potent than S-14 (EC(50) 0.04 vs. 13.2 nM). PYY was inhibited by two analogs with affinity for SSTR-5, BIM-23268 and BIM-23052, more potently than S-14 and as effectively as S-28. The SSTR-5 analog L-362855 suppressed PYY equivalent only to S-14, but the structurally related peptide L-372588 (Phe to Tyr at position 2) was equipotent to S-28, whereas L-372587 (Phe to Tyr at position 7) caused no inhibition. An SSTR-2 analog decreased PYY secretion similar to S-14, and an SSTR 3 analog was ineffective. PYY secretion stimulated by phorbol 12-myristate 13 acetate and by forskolin was also more potently suppressed by S-28 and the octapeptide SSTR-5 analogs. The results indicate that S-28 mediates inhibition of gastrin-releasing peptide-stimulated PYY secretion through activation of SSTR-5 and includes suppression of cAMP- and protein kinase C-dependent pathways. Substitution of a single hydroxyl group confers differences in SSTR-5 agonist properties, suggesting region specificity for the intrinsic activity of this receptor subtype. PMID- 11052996 TI - Effect of splanchnectomy on jejunal motility and fos expression in brain stem after intestinal anaphylaxis in rat. AB - This study was to determine whether alterations in jejunal motility observed after antigen challenge of sensitized rats occurred after extirpation of the celiac-superior mesenteric ganglia. Hooded-Lister rats were prepared with an intact or extirpated celiac-superior mesenteric ganglion, an isolated Thiry-Vella loop of ileum for instillation of antigen, and jejunal electrodes for myoelectric recording. Animals were sensitized by injection of 10 microg egg albumin (EA, ip), and specific anti-EA IgE titers were determined to be >1:64. In both control and splanchnectomized rats, normal fasting migrating myoelectric complexes (MMC) were observed before challenge with EA. MMCs were disrupted, and diarrhea was observed immediately after EA challenge of control but not splanchnectomized animals. Brain stems were removed and processed for Fos immunoreactivity. The absence of perivascular neuropeptide Y immunoreactivity in the submucosa was used to confirm the success of splanchnectomy. The number of Fos-immunoreactive neuronal nuclei was significantly reduced in the brain stem after splanchnectomy. Thus the mesenteric sympathetic ganglia are an integral part of the extramural neuronal pathways required for altered motility in this model of intestinal anaphylaxis. PMID- 11052997 TI - Expression of cell volume-regulated kinase h-sgk in pancreatic tissue. AB - Transcript levels of the human serine/threonine kinase h-sgk have been found to be highest in pancreas. In the present study, localization and regulation of h sgk transcription in pancreatic tissue were elucidated. As was apparent from radioactive in situ hybridization, most pancreatic acinar cells expressed high levels of h-sgk mRNA. h-sgk mRNA-positive cells were also found in ductal epithelia but not in pancreatic islets. In biopsy specimens from patients with pancreatitis, h-sgk mRNA levels were decreased in acinar cells but abundant in numerous mononuclear interstitial cells within areas of pancreatic necrosis and fibrosis. As shown by Northern blotting, h-sgk transcription in DAN-G pancreatic tumor cells is upregulated by osmotic cell shrinkage, serum, phorbol esters (phorbol 12,13-didecanoate), and Ca(2+) ionophore A-23187 and decreased by staurosporine and cAMP. In conclusion, h-sgk transcription is regulated not only by cell volume but also by serum, protein kinase C stimulation, cAMP, and increase of intracellular Ca(2+) activity. The kinase may participate not only in normal function of exocrine pancreas but also in fibrosing pancreatitis. PMID- 11052998 TI - Novel goblet cell gene related to IgGFcgammaBP is regulated in adapting gut after small bowel resection. AB - The loss of functional small bowel surface area leads to a well-described adaptive response in the remnant intestine. To elucidate its molecular regulation, a cohort of cDNAs were cloned using a rat gut resection model and subtractive/differential hybridization cloning techniques. This study reports a novel cDNA termed "ileal remnant repressed" (IRR)-219, which shares 80% nucleotide identity with the 3'end of a human intestinal IgG Fc binding protein (IgGFcgammaBP) and is homologous to human and rat mucins. IRR-219 mRNA is expressed in intestine and colon only. At 48 h after 70% intestinal resection, mRNA levels decreased two- to fivefold in the adaptive small bowel but increased two- to threefold in the colon. Expression of IRR-219 was suppressed in adaptive small bowel as late as 1 wk after resection. IRR-219 expression is also regulated during gut ontogeny. In situ hybridization revealed IRR-219 expression in small intestinal and colonic goblet cells only. Its unique patterns of expression during ontogeny and after small bowel resection suggest distinctive roles in small bowel and colonic adaptation. PMID- 11052999 TI - Keratinocyte growth factor-2 (FGF-10) promotes healing of experimental small intestinal ulceration in rats. AB - Keratinocyte growth factor-2 (KGF-2, repifermin) is a homolog of KGF-1 with epithelial mitogenic activities. We investigated the therapeutic role of KGF-2 in intestinal ulceration and its mechanisms of protection. KGF-2 (0.3-5 mg/kg) was administered before or after induction of small intestinal ulceration by indomethacin (Indo) in prevention and treatment protocols. In acute studies, KGF 2 was injected for up to 7 days before or daily for 5 days after Indo. In a 15 day chronic study, KGF-2 was injected intravenously daily beginning before or 7 days after Indo. Injury was evaluated by blinded macroscopic and microscopic inflammatory scores, epithelial BrdU staining, tissue IL-1beta, PGE(2), and hydroxyproline concentrations, and collagen type I RNA expression. In vitro effects of KGF-2 were evaluated by epithelial cellular proliferation, restitution of wounded monolayers, PGE(2) secretion, and expression of COX-2 and collagen mRNA. Intravenous KGF-2 significantly decreased acute intestinal injury by all parameters and significantly decreased chronic ulceration. Pretreatment, daily infusion, and delayed treatment were effective. KGF-2 promoted in vitro epithelial restitution with only modest effects on epithelial cell proliferation, stimulated COX-2 expression in cultured epithelial cells, and upregulated in vitro and in vivo PGE(2) production. KGF-2 did not affect in vivo fibrosis, although it induced collagen expression in cultured intestinal myofibroblasts. These results suggest that KGF-2 inhibits intestinal inflammation by stimulating epithelial restitution and protective PGs. PMID- 11053000 TI - Gene transfer of recombinant endothelial nitric oxide synthase to liver in vivo and in vitro. AB - Endothelial nitric oxide synthase (eNOS)-derived nitric oxide (NO) contributes to hepatic vascular homeostasis. The aim of this study was to examine whether delivery of an adenoviral vector encoding eNOS gene to liver affects vasomotor function in vivo and the mechanism of NO production in vitro. Rats were administered adenoviruses encoding beta-galactosidase (AdCMVLacZ) or eNOS (AdCMVeNOS) via tail vein injection and studied 1 wk later. In animals transduced with AdCMVLacZ, beta-galactosidase activity was increased in the liver, most prominently in hepatocytes. In AdCMVeNOS-transduced animals, eNOS protein levels and catalytic activity were significantly increased. Overexpression of eNOS diminished baseline perfusion pressure and constriction in response to the alpha(1)-agonist methoxamine in the perfused liver. Transduction of cultured hepatocytes with AdCMVeNOS resulted in the targeting of recombinant eNOS to a perinuclear distribution and binding with the NOS-activating protein heat shock protein 90. These events were associated with increased ionomycin-stimulated NO release. In summary, this is the first study to demonstrate successful delivery of the recombinant eNOS gene to liver in vivo and in vitro with ensuing NO production. PMID- 11053001 TI - Adenovirus-mediated human pancreatic lipase gene transfer to rat bile: gene therapy of fat malabsorption. AB - This study explored the potential of using the gene therapy approach, based on adenovirus-mediated expression of pancreatic lipase in the hepatobiliary tract, to increase lipid digestion in the intestinal lumen and promote lipid absorption through the gastrointestinal tract. Recombinant adenovirus containing the human pancreatic lipase cDNA (AdPL) was shown to transduce and mediate pancreatic lipase biosynthesis in rat IEC-6 epithelial cells in vitro. Retrograde infusion of recombinant adenovirus (3 x 10(8) plaque-forming units) containing the bacterial LacZ gene (AdLacZ) into the bile duct of rats resulted in positive X gal reaction products in the periportal liver cells 7 days after AdLacZ infusion. A high level of human pancreatic lipase was detected in bile after retrograde bile duct infusion of rats with AdPL but not in the bile of animals infused with AdLacZ. Triglyceride hydrolytic activity in the bile of AdPL-infused rats was equivalent to that present in pancreatic juice. In contrast, serum obtained from these animals did not contain any detectable pancreatic lipase activity. These results suggest that ectopic expression of pancreatic enzymes in the hepatobiliary tract may be an alternative therapeutic strategy for treating fat malabsorption due to pancreatic insufficiency. PMID- 11053002 TI - Selective sparing of goblet cells and paneth cells in the intestine of methotrexate-treated rats. AB - Proliferation, differentiation, and cell death were studied in small intestinal and colonic epithelia of rats after treatment with methotrexate. Days 1-2 after treatment were characterized by decreased proliferation, increased apoptosis, and decreased numbers and depths of small intestinal crypts in a proximal-to-distal decreasing gradient along the small intestine. The remaining crypt epithelium appeared flattened, except for Paneth cells, in which lysozyme protein and mRNA expression was increased. Regeneration through increased proliferation during days 3-4 coincided with villus atrophy, showing decreased numbers of villus enterocytes and decreased expression of the enterocyte-specific genes sucrase isomaltase and carbamoyl phosphate synthase I. Remarkably, goblet cells were spared at villus tips and remained functional, displaying Muc2 and trefoil factor 3 expression. On days 8-10, all parameters had returned to normal in the whole small intestine. No methotrexate-induced changes were seen in epithelial morphology, proliferation, apoptosis, Muc2, and TFF3 immunostaining in the colon. The observed small intestinal sparing of Paneth cells and goblet cells following exposure to methotrexate is likely to contribute to epithelial defense during increased vulnerability of the intestinal epithelium. PMID- 11053003 TI - Regulation of parietal cell calcium signaling in gastric glands. AB - The ligands interacting with enterochromaffin-like (ECL) and parietal cells and the signaling interactions between these cells were investigated in rabbit gastric glands using confocal microscopy. Intracellular calcium concentration ([Ca(2+)](i)) changes were used to monitor cellular responses. Histamine and carbachol increased [Ca(2+)](i) in parietal cells. Gastrin (1 nM) increased [Ca(2+)](i) in ECL cells and adjacent parietal cells. Only the increase of [Ca(2+)](i) in parietal cells was inhibited by H(2) receptor antagonists (H(2)RA). Gastrin (10 nM) evoked an H(2)RA-insensitive [Ca(2+)](i) increase in parietal cells. Carbachol produced large H(2)RA- and somatostatin-insensitive signals in parietal cells. Pituitary adenylate cyclase-activating peptide (PACAP, 100 nM) elevated [Ca(2+)](i) in ECL cells and adjacent parietal cells. H(2)RAs abolished the PACAP-stimulated [Ca(2+)](i) increase in adjacent parietal cells. Somatostatin did not inhibit the increase of [Ca(2+)](i) in parietal cells stimulated with histamine, high gastrin concentrations, or carbachol but abolished ECL cell calcium responses to gastrin or PACAP. Hence, rabbit parietal cells express histaminergic, muscarinic, and CCK-B receptors coupled to calcium signaling but insensitive to somatostatin, whereas rabbit and rat ECL cells express PACAP and CCK-B calcium coupled receptors sensitive to somatostatin. PMID- 11053004 TI - Human esophageal smooth muscle cells express muscarinic receptor subtypes M(1) through M(5). AB - Receptor characterization in human esophageal smooth muscle is limited by tissue availability. We used human esophageal smooth muscle cells in culture to examine the expression and function of muscarinic receptors. Primary cultures were established using cells isolated by enzymatic digestion of longitudinal muscle (LM) and circular muscle (CM) obtained from patients undergoing esophagectomy for cancer. Cultured cells grew to confluence after 10-14 days in medium containing 10% fetal bovine serum and stained positively for anti-smooth muscle specific alpha-actin. mRNA encoding muscarinic receptor subtypes M(1)-M(5) was identified by RT-PCR. The expression of corresponding protein for all five subtypes was confirmed by immunoblotting and immunocytochemistry. Functional responses were assessed by measuring free intracellular Ca(2+) concentration ([Ca(2+)](i)) using fura 2 fluorescence. Basal [Ca(2+)](i), which was 135 +/- 22 nM, increased transiently to 543 +/- 29 nM in response to 10 microM ACh in CM cells (n = 8). This response was decreased <95% by 0.01 microM 4-diphenylacetoxy-N methylpiperidine, a M(1)/M(3)-selective antagonist, whereas 0.1 microM methoctramine, a M(2)/M(4)-selective antagonist, and 0.1 microM pirenzepine, a M(1)-selective antagonist, had more modest effects. LM and CM cells showed similar results. We conclude that human smooth muscle cells in primary culture express five muscarinic receptor subtypes and respond to ACh with a rise in [Ca(2+)](i) mediated primarily by the M(3) receptor and involving release of Ca(2+) from intracellular stores. This culture model provides a useful tool for further study of esophageal physiology. PMID- 11053005 TI - Dietary iron induces rapid changes in rat intestinal divalent metal transporter expression. AB - The divalent metal transporter (DMT1, also known as NRAMP2 or DCT1) is the likely target for regulation of intestinal iron absorption by iron stores. We investigated changes in intestinal DMT1 expression after a bolus of dietary iron in iron-deficient Belgrade rats homozygous for the DMT1 G185R mutation (b/b) and phenotypically normal heterozygous littermates (+/b). Immunofluorescent staining with anti-DMT1 antisera showed that DMT1 was located in the brush-border membrane. Duodenal DMT1 mRNA and protein levels were six- and twofold higher, respectively, in b/b rats than in +/b rats. At 1.5 h after dietary iron intake in +/b and b/b rats, DMT1 was internalized into cytoplasmic vesicles. At 1.5 and 3 h after iron intake in +/b and b/b rats, there was a rapid decrease of DMT1 mRNA and a transient increase of DMT1 protein. The decrease of DMT1 mRNA was specific, because ferritin mRNA was unchanged. After iron intake, an increase in ferritin protein and decrease in iron-regulatory protein binding activity occurred, reflecting elevated intracellular iron pools. Thus intestinal DMT1 rapidly responds to dietary iron in both +/b and b/b rats. The internalization of DMT1 may be an acute regulatory mechanism to limit iron uptake. In addition, the results suggest that in the Belgrade rat DMT1 with the G185R mutation is not an absolute block to iron. PMID- 11053006 TI - A duodenum-specific enhancer regulates expression along three axes in the small intestine. AB - Adenosine deaminase (ADA) is expressed at high levels in the epithelium of proximal small intestine. Transgenic mice were used to characterize the regulatory region governing this activation. A duodenum-specific enhancer is located in intron 2 of the human ADA gene at the central site among a cluster of seven DNase I-hypersensitive sites present in duodenal DNA. Flanking DNA, including the remaining hypersensitive sites, is required for consistent high level enhancer function. The enhancer activates expression in a pattern identical to endogenous ADA along both the anterior-posterior axis of the small intestine and the crypt-villus differentiation axis of the intestinal epithelium. Timing of activation by the central enhancer mimics endogenous mouse ADA activation, occurring at 2-3 wk of age. However, two upstream DNA segments, one proximal and one distal, collaborate to change enhancer activation to a perinatal time point. Studies with duodenal nuclear extracts identified five distinct DNase I footprints within the enhancer. Protected regions encompass six putative binding sites for the transcription factor PDX-1, as well as proposed CDX, hepatocyte nuclear factor-4, and GATA-type sites. PMID- 11053007 TI - Superantigen immune stimulation activates epithelial STAT-1 and PI 3-K: PI 3-K regulation of permeability. AB - Signal transducers and activators of transcription (STATs) are critical intracellular signaling molecules for many cytokines. We compared the ability of T84 epithelial cells to activate STATs in response to cytokines [interferon-gamma (IFN-gamma), interleukin (IL)-4, IL-10, and tumor necrosis factor-alpha (10 ng/ml)] and conditioned medium from superantigen [Staphylococcus aureus enterotoxin B (SEB)]-activated peripheral blood mononuclear cells (PBMC) using electrophoretic mobility shift assays (EMSA). Of the cytokines tested, only IFN gamma caused a STAT-1 response. Exposure to SEB-PBMC-conditioned medium resulted in STAT-1 or STAT-1/3 activation, and inclusion of anti-IFN-gamma antibodies in the conditioned medium abolished the STAT-1 signal. Cells treated with transcription factor decoys, DNA oligonucleotides bearing the STAT-1 recognition motif, and then SEB-PBMC-conditioned medium displayed a reduced STAT-1 signal on EMSA, yet this treatment did not prevent the drop in transepithelial resistance (measured in Ussing chambers) caused by SEB-PBMC-conditioned medium. In contrast, the phosphatidylinositol 3'-kinase (PI 3-K) inhibitor LY-294002 significantly reduced the drop in transepithelial resistance caused by SEB-PBMC-conditioned medium. Thus data are presented showing STAT-1 (+/-STAT-3) and PI 3-K activation in epithelial cells in response to immune mediators released by superantigen immune activation. Although the involvement of STAT-1/-3 in the control of barrier function remains a possibility, PI-3K has been identified as a regulator of T84 paracellular permeability. PMID- 11053008 TI - Inhibition of enterotoxin-induced porcine colonic secretion by diarylsulfonylureas in vitro. AB - Muscle-stripped piglet colon was used to evaluate changes in short-circuit current (I(sc)) as an indicator of anion secretion. Mucosal exposure to Escherichia coli heat-stable (STa) or heat-labile enterotoxins (LT) stimulated I(sc) by 32 +/- 5 and 42 +/- 7 microA/cm(2), respectively. Enterotoxin-stimulated I(sc) was not significantly affected by either 4,4'-diaminostilbene-2, 2' disulfonic acid or CdCl(2), inhibitors of Ca(2+)-activated Cl(-) channels and ClC 2 channels, respectively. Alternatively, N-(4-methylphenylsulfonyl)-N'-(4 trifluoromethylphenyl)urea (DASU-02), a compound that inhibits cystic fibrosis transmembrane conductance regulator (CFTR)-mediated Cl(-) secretion, reduced I(sc) by 29 +/- 7 and 34 +/- 11 microA/cm(2), respectively. Two additional diarylsulfonylurea (DASU)-based compounds were evaluated for their effects on enterotoxin-stimulated secretion. The rank order of potency for inhibition by these three closely related DASU structures was identical to that observed for human CFTR. The degree of inhibition by each of these compounds was similar for both STa and LT. The structure- and concentration-dependent inhibition shown indicates that CFTR mediates both STa- and LT-stimulated colonic secretion. Similar structure-dependent inhibitory effects were observed in forskolin stimulated rat colonic epithelium. Thus DASUs compose a family of inhibitors that may be of therapeutic value for the symptomatic treatment of diarrhea resulting from a broad spectrum of causative agents across species. PMID- 11053009 TI - Time course inhibition of gastric and platelet COX activity by acetylsalicylic acid in humans. AB - Aspirin causes peptic ulcers predominately by reducing gastric mucosal cyclooxygenase (COX) activity and prostaglandin synthesis. Because aspirin circulates for only a few hours, we hypothesized that aspirin's inhibitory effect on gastric COX activity must be prolonged. We performed a placebo-controlled experiment in healthy humans to determine the duration of inhibition of aspirin on gastric mucosal COX activity (PGE(2) and PGF(2alpha) synthesis rates). Recovery of gastric COX activity after stopping aspirin was slow and linear. Seventy-two hours after 325-mg aspirin, gastric COX activity was still reduced by 57% (P < 0.001). Duration of inhibition of gastric COX activity was estimated to be 7-8 days after 325-mg aspirin and 5 days after 81-mg aspirin. Recovery of gastric prostaglandin synthesis after 325-mg but not after 81-mg aspirin occurred at slower rates in subjects with Helicobacter pylori-associated gastritis than in those with normal histology. In conclusion, aspirin inhibits gastric COX activity for much longer than predicted from its pharmacokinetic profile, explaining why aspirin at widely spaced intervals is ulcerogenic. PMID- 11053010 TI - Differential Ca(2+) signaling characteristics of inhibitory and excitatory myenteric motor neurons in culture. AB - Physiological studies on functionally identified myenteric neurons are scarce because of technical limitations. We combined retrograde labeling, cell culturing, and fluorescent intracellular Ca(2+) concentration ([Ca(2+)](i)) signaling to study excitatory neurotransmitter responsiveness of myenteric motor neurons. 1, 1-Didodecyl-3,3,3',3'-tetramethyl indocarbocyanine (DiI) was used to label circular muscle motor neurons of the guinea pig ileum. DiI-labeled neurons were easily detectable in cultures prepared from these segments. The excitatory neurotransmitters (10(-5) M) acetylcholine, substance P, and serotonin induced a transient rise in [Ca(2+)](i) in subsets of DiI-labeled neurons (66.7, 56.5, and 84. 3%, respectively). DiI-labeled motor neurons were either inhibitory (23.8%) or excitatory (76.2%) as assessed by staining for nitric oxide synthase or choline acetyltransferase. Compared with excitatory motor neurons, significantly fewer inhibitory neurons in culture responded to acetylcholine (0 vs. 69%) and substance P (12.5 vs. 69.2%). We conclude that combining retrograde labeling and Ca(2+) imaging allows identification of differential receptor expression in functionally identified neurons in culture. PMID- 11053011 TI - Structural determinants of antiproliferative activity of heparin on pulmonary artery smooth muscle cells. AB - In addition to its anticoagulant properties, heparin (HP), a complex polysaccharide covalently linked to a protein core, inhibits proliferation of several cell types including pulmonary artery smooth muscle cells (PASMCs). Commercial lots of HP exhibit varying degrees of antiproliferative activity on PASMCs that may due to structural differences in the lots. Fractionation of a potent antiproliferative HP preparation into high and low molecular weight components does not alter the antiproliferative effect on PASMCs, suggesting that the size of HP is not the major determinant of this biological activity. The protein core of HP obtained by cleaving the carbohydrate-protein linkage has no growth inhibition on PASMCs, demonstrating that the antiproliferative activity resides in the glycosaminoglycan component. Basic sugar residues of glucosamine can be replaced with another basic sugar, i.e., galactosamine, without affecting growth inhibition of PASMCs. N-sulfonate groups on these sugar residues of HP are not essential for growth inhibition. However, O-sulfonate groups on both sugar residues are essential for the antiproliferative activity on PASMCs. In whole HP, in contrast to an earlier finding based on a synthetic pentasaccharide of HP, 3-O sulfonation is not critical for the antiproliferative activity against PASMCs. The amounts and distribution of sulfonate groups on both sugar residues of the glycosaminoglycan chain are the major determinant of antiproliferative activity. PMID- 11053012 TI - Complement-mediated host defense in the lung. AB - Complement is a system of plasma proteins that aids in the elimination of pathogens from the body. We hypothesized that there is a functional complement system present in the lung that aids in the removal of pathogens. Western blot analysis revealed complement proteins of the alternative and classical pathways of complement in bronchoalveolar lavage fluids (BALF) from healthy volunteers. Functional classical pathway activity was detected in human BALF, but there was no significant alternative pathway activity in lavage fluid, a finding that correlates with the low level of the alternative pathway protein, factor B, in these samples. Although the classical pathway of complement was functional in lavage fluid, the level of the classical pathway activator C1q was very low. We tested the ability of the lung- specific surfactant proteins, surfactant protein A (SP-A) and surfactant protein D (SP-D), to substitute for C1q in classical pathway activation, since they have structural homology to C1q. However, neither SP-A nor SP-D restored classical pathway activity to C1q-depleted serum. These data suggest that the classical pathway of complement is functionally active in the lung where it may play a role in the recognition and clearance of bacteria. PMID- 11053013 TI - Synergistic and additive killing by antimicrobial factors found in human airway surface liquid. AB - Airway surface liquid contains multiple factors thought to provide a first line of defense against bacteria deposited in the airways. Although the antimicrobial action of individual factors has been studied, less is known about how they work in combination. We examined the combined action of six antimicrobial peptides found in airway surface liquid. The paired combinations of lysozyme-lactoferrin, lysozyme-secretory leukocyte protease inhibitor (SLPI), and lactoferrin-SLPI were synergistic. The triple combination of lysozyme, lactoferrin, and SLPI showed even greater synergy. Other combinations involving the human beta-defensins, LL 37, and tobramycin (often administered to cystic fibrosis patients by inhalation) were additive. Because the airway surface liquid salt concentration may be elevated in cystic fibrosis patients, we examined the effect of salt on the synergistic combinations. As the ionic strength increased, synergistic interactions were lost. Our data suggest that the antibacterial potency of airway surface liquid may be significantly increased by synergistic and additive interactions between antimicrobial factors. These results also suggest that increased salt concentrations that may exist in cystic fibrosis could inhibit airway defenses by diminishing these synergistic interactions. PMID- 11053014 TI - Characterization of rabbit SP-B promoter region responsive to downregulation by tumor necrosis factor-alpha. AB - Surfactant protein B (SP-B) is essential for the maintenance of biophysical properties and physiological function of pulmonary surfactant. Tumor necrosis factor-alpha (TNF-alpha), an important mediator of lung inflammation, inhibits surfactant phospholipid and surfactant protein synthesis in the lung. In the present study, we investigated the TNF-alpha inhibition of rabbit SP-B promoter activity in a human lung adenocarcinoma cell line (NCI-H441). Deletion experiments indicated that the TNF-alpha response elements are located within 236 bp of SP-B 5'-flanking DNA. The TNF-alpha response region contained binding sites for nuclear factor-kappa B (NF-kappa B), Sp1/Sp3, thyroid transcription factor (TTF)-1, and hepatocyte nuclear factor (HNF)-3 transcription factors. Inhibitors of NF-kappa B activation such as dexamethasone and N-tosyl-L phenylalanine chloromethyl ketone and mutation of the NF-kappa B element did not reverse TNF-alpha inhibition of SP-B promoter, indicating that TNF-alpha inhibition of SP-B promoter activity occurs independently of NF-kappa B activation. TNF-alpha treatment decreased the binding activities of TTF-1 and HNF 3 elements without altering the nuclear levels of TTF-1 and HNF-3 alpha proteins. Pretreatment of cells with okadaic acid reversed TNF-alpha inhibition of SP-B promoter activity. Taken together these data indicated that in NCI-H441 cells 1) TNF-alpha inhibition of SP-B promoter activity may be caused by decreased binding activities of TTF-1 and HNF-3 elements, 2) the decreased binding activities of TTF-1 and HNF-3 alpha are not due to decreased nuclear levels of the proteins, and 3) okadaic acid-sensitive phosphatases may be involved in mediating TNF-alpha inhibition of SP-B promoter activity. PMID- 11053015 TI - Store-operated calcium entry and increased endothelial cell permeability. AB - We hypothesized that myosin light chain kinase (MLCK) links calcium release to activation of store-operated calcium entry, which is important for control of the endothelial cell barrier. Acute inhibition of MLCK caused calcium release from inositol trisphosphate-sensitive calcium stores and prevented subsequent activation of store-operated calcium entry by thapsigargin, suggesting that MLCK serves as an important mechanism linking store depletion to activation of membrane calcium channels. Moreover, in voltage-clamped single rat pulmonary artery endothelial cells, thapsigargin activated an inward calcium current that was abolished by MLCK inhibition. F-actin disruption activated a calcium current, and F-actin stabilization eliminated the thapsigargin-induced current. Thapsigargin increased endothelial cell permeability in the presence, but not in the absence, of extracellular calcium, indicating the importance of calcium entry in decreasing barrier function. Although MLCK inhibition prevented thapsigargin from stimulating calcium entry, it did not prevent thapsigargin from increasing permeability. Rather, inhibition of MLCK activity increased permeability that was especially prominent in low extracellular calcium. In conclusion, MLCK links store depletion to activation of a store-operated calcium entry channel. However, inhibition of calcium entry by MLCK is not sufficient to prevent thapsigargin from increasing endothelial cell permeability. PMID- 11053016 TI - Hypoxia modifies the effect of PDGF on glycosaminoglycan synthesis by primary human lung cells. AB - Hypoxia, a consequence of interstitial lung diseases, may lead to secondary pulmonary hypertension and pulmonary vascular remodeling. Hypoxia induces activation and proliferation of lung cells and enhances the deposition of extracellular matrix including glycosaminoglycans (GAGs). To elucidate the cell biological mechanisms underlying the development of secondary pulmonary hypertension, we studied the effect of hypoxia on GAG synthesis by human lung cells. GAG synthesis was measured by incorporation of [(3)H]glucosamine; GAGs were isolated, purified, and characterized with GAG-degrading enzymes. Fibroblasts and vascular smooth muscle cells (VSMCs) synthesized hyaluronic acid, heparan sulfate, and chondroitin sulfates, whereas dermatan sulfate was found only in fibroblasts. Hypoxia did not influence the size or charge of the individual GAGs. However, hypoxia inhibited platelet-derived growth factor induced [(3)H]glucosamine incorporation in secreted GAGs, especially hyaluronic acid, in VSMCs. In contrast, it stimulated GAG secretion, specifically heparan sulfate, by fibroblasts. Our results indicate that hypoxia induces modifications in GAG synthesis by human lung VSMCs and fibroblasts that may be correlated to pathophysiological manifestations in lung diseases causing hypoxia. PMID- 11053017 TI - Contribution of R domain phosphoserines to the function of CFTR studied in Fischer rat thyroid epithelia. AB - The regulatory domain of cystic fibrosis transmembrane conductance regulator (CFTR) regulates channel activity when several serines are phosphorylated by cAMP dependent protein kinase. To further define the functional role of individual phosphoserines, we studied CFTR containing previously studied and new serine to alanine mutations. We expressed these constructs in Fischer rat thyroid epithelia and measured transepithelial Cl(-) current. Mutation of four in vivo phosphorylation sites, Ser(660), Ser(737), Ser(795), and Ser(813) (S-Quad-A), substantially decreased cAMP-stimulated current, suggesting that these four sites account for most of the phosphorylation-dependent response. Mutation of either Ser(660) or Ser(813) alone significantly decreased current, indicating that these residues play a key role in phosphorylation-dependent stimulation. However, neither Ser(660) nor Ser(813) alone increased current to wild-type levels; both residues were required. Changing Ser(737) to alanine increased current above wild type levels, suggesting that phosphorylation of Ser(737) may inhibit current in wild-type CFTR. These data help define the functional role of regulatory domain phosphoserines and suggest interactions between individual phosphoserines. PMID- 11053018 TI - Surfactant phospholipid catabolic rate is pool size dependent in mice. AB - We increased surfactant pool size by surfactant treatment in mice to test if the catabolism of the major component of surfactant, saturated phosphatidylcholine (Sat PC), was rate limited. By intratracheal instillation, we gave mice trace doses, doses of 45 or 110 micromol/kg, or three doses of 110 micromol/kg of Sat PC in surfactant that contained radiolabeled dipalmitoylphosphatidylcholine (DPPC) and a radiolabeled phospholipase A-resistant ether analog of DPPC. Two strains of mice with 2-fold differences in alveolar and total Sat PC pool sizes were used; the mice with the higher pool sizes had a 2.3-fold higher steady-state catabolic rate. Acute increases in alveolar surfactant given by intratracheal instillation increased catabolic rates approximately 2-fold over the steady-state rates in both strains. There was minimal loss of the ether analog of DPPC from the lungs, and the alveolar macrophages did not accumulate more than 10% of the ether analog. In these two strains of mice, the catabolism of Sat PC was not rate limited because catabolic rate increased when alveolar pool sizes were increased. PMID- 11053019 TI - Angiotensin IV-mediated pulmonary artery vasorelaxation is due to endothelial intracellular calcium release. AB - Angiotensin (ANG) IV stimulation of pulmonary artery (PA) endothelial cells (PAECs) but not of PA smooth muscle cells (PASMCs) resulted in significant increased production of cGMP in PASMCs. ANG IV receptors are not present in PASMCs, and PASMC nitric oxide synthase activity was not altered by ANG IV. ANG IV caused a dose-dependent vasodilation of U-46619-precontracted endothelium intact but not endothelium-denuded PAs, and this response was blocked by the ANG IV receptor antagonist divalinal ANG IV but not by ANG II type 1 and 2 receptor blockers. ANG IV receptor-mediated increased intracellular Ca(2+) concentration ([Ca(2+)](i)) release from intracellular stores in PAECs was blocked by divalinal ANG IV as well as by the G protein, phospholipase C, and phosphoinositide (PI) 3 kinase inhibitors guanosine 5'-O-(2-thiodiphosphate), U-73122, and LY-294002, respectively, and was regulated by both PI 3-kinase- and ryanodine-sensitive Ca(2+) stores. Basal and ANG IV-mediated vasorelaxation of endothelium-denuded PAs was restored by exogenous PAECs but not by exogenous PAECs pretreated with the intracellular Ca(2+) chelator 1,2-bis(o-aminophenoxy)ethane-N,N,N',N' tetraacetic acid-AM. These results demonstrate that ANG IV-mediated vasodilation of PAs is endothelium dependent and regulated by [Ca(2+)](i) release through receptor-coupled G protein-phospholipase C-PI 3-kinase signaling mechanisms. PMID- 11053020 TI - Chronic intrauterine pulmonary hypertension decreases calcium-sensitive potassium channel mRNA expression. AB - Calcium-sensitive potassium (K(Ca)) channels play a critical role in mediating perinatal pulmonary vasodilation. Because infants with persistent pulmonary hypertension of the newborn (PPHN) have blunted vasodilator responses to birth related stimuli, we hypothesized that lung K(Ca) channel gene expression is decreased in PPHN. To test this hypothesis, we measured K(Ca) channel gene expression in distal lung homogenates from both fetal lambs with severe pulmonary hypertension caused by prolonged compression of the ductus arteriosus and age matched, sham-operated animals (controls). After at least 9 days of compression of the ductus arteriosus, fetal lambs were killed. To determine lung K(Ca) channel mRNA levels, primers were designed against the known sequence of the K(Ca) channel and used in semiquantitative RT-PCR, with lung 18S rRNA content as an internal control. Compared to that in control lambs, lung K(Ca) channel mRNA content in the PPHN group was reduced by 26 +/- 6% (P < 0.02), whereas lung voltage-gated K(+) 2.1 mRNA content was unchanged. We conclude that lung K(Ca) channel mRNA expression is decreased in an ovine model of PPHN. Decreased K(Ca) channel gene expression may contribute to the abnormal pulmonary vascular reactivity associated with PPHN. PMID- 11053021 TI - Hoxa-5 in mouse developing lung: cell-specific expression and retinoic acid regulation. AB - Hoxa-5 is a homeobox gene that is highly expressed in the developing mouse lung. However, little is known about the molecular mechanisms controlling expression. We characterized the ontogeny of Hoxa-5 gene and protein expressions during lung development and then studied the cell-specific effects of retinoic acid (RA) on Hoxa-5 mRNA in fetal lung fibroblasts and MLE-12 mouse lung epithelial cells. Strong but constant Hoxa-5 gene and protein expressions were detected from mouse lung on embryonic day 13.5 to postnatal day 2. At baseline, the gene was strongly expressed in the fibroblasts of day 17.5 fetal mouse lungs. A very weak but reproducible expression was present in the MLE-12 cells. RA stimulated gene expression in both cell types in a time- and dose-dependent manner. Peak expression occurred much later in the MLE-12 cells compared with that in fibroblasts. Cycloheximide and actinomycin D treatment studies suggested that the differences in RA effect on each cell type may involve the presence of a repressor that can be overcome by RA. PMID- 11053022 TI - Inhibition of endotoxin-induced lung inflammation by interleukin-10 gene transfer in mice. AB - Interleukin (IL)-10 is an anti-inflammatory cytokine that has great potential for use in the treatment of inflammatory and immune illnesses. In this study, gene transfer was used to induce IL-10 transgene expression in murine lungs for treatment of endotoxin-induced lung inflammation. Gene transfer was performed with a cytomegalovirus (CMV)-IL-10 plasmid with the aid of the liposomal agents LipofectAMINE and N-[1-(2,3-dioleoyl)propyl]-N,N, N-trimethylammonium methylsulfate (DOTAP). Administration of the endotoxin caused a marked increase in lung inflammation as indicated by increased tumor necrosis factor (TNF)-alpha release and neutrophil count. Pretreatment of the mice with IL-10 plasmid with and without LipofectAMINE had no inhibitory effect on lung inflammation and IL-10 transgene expression. LipofectAMINE by itself induced lung inflammation, an effect that was not observed with DOTAP. IL-10 plasmid when codelivered with DOTAP expressed biologically active IL-10 protein and caused a reduction in endotoxin-induced inflammation. Transgene expression was observed as early as 3 h after administration, peaked at 12 h, and declined thereafter. We conclude that IL-10 gene transfer is a feasible approach for the treatment of lung inflammation. PMID- 11053023 TI - Free radical-mediated transgene inactivation of macrophages by endotoxin. AB - Endotoxin, the lipopolysaccharide component of gram-negative bacteria, is a common contaminant of plasmid DNA preparations. The present study investigated the effect of endotoxin on gene transfection efficiency and the role of reactive oxygen species (ROS) in this process. Gene transfection studies were performed in various cell types with cytomegalovirus-luciferase as a reporter plasmid and cationic liposome as a transfecting agent. The presence of endotoxin in plasmid DNA preparations severely limited transgene expression in macrophages but had little or no effect in other cell types tested. This decreased transfection was dependent on ROS-mediated cellular toxicity induced by endotoxin. Neutralizing the endotoxin by the addition of polymyxin B effectively increased transfection efficiency and reduced toxicity. Electron spin resonance studies confirmed the formation of ROS in endotoxin-treated cells and their inhibition by free radical scavengers. The ROS scavenger N-t-butyl-alpha-phenylnitrone, the H(2)O(2) scavenger catalase, and the.OH scavenger sodium formate effectively inhibited endotoxin-induced effects, whereas the O(2)(-) scavenger superoxide dismutase had lesser effects. These results indicate that multiple oxidative species are involved in the transfection inactivation process and that.OH formed by H(2)O(2) dependent, metal-catalyzed Fenton reaction play a major role in this process. PMID- 11053024 TI - L-type Ca(2+) channels, resting [Ca(2+)](i), and ET-1-induced responses in chronically hypoxic pulmonary myocytes. AB - In the lung, chronic hypoxia (CH) causes pulmonary arterial smooth muscle cell (PASMC) depolarization, elevated endothelin-1 (ET-1), and vasoconstriction. We determined whether, during CH, depolarization-driven activation of L-type Ca(2+) channels contributes to 1) maintenance of resting intracellular Ca(2+) concentration ([Ca(2+)](i)), 2) increased [Ca(2+)](i) in response to ET-1 (10(-8) M), and 3) ET-1-induced contraction. Using indo 1 microfluorescence, we determined that resting [Ca(2+)](i) in PASMCs from intrapulmonary arteries of rats exposed to 10% O(2) for 21 days was 293.9 +/- 25.2 nM (vs. 153.6 +/- 28.7 nM in normoxia). Resting [Ca(2+)](i) was decreased after extracellular Ca(2+) removal but not with nifedipine (10(-6) M), an L-type Ca(2+) channel antagonist. After CH, the ET-1-induced increase in [Ca(2+)](i) was reduced and was abolished after extracellular Ca(2+) removal or nifedipine. Removal of extracellular Ca(2+) reduced ET-1-induced tension; however, nifedipine had only a slight effect. These data indicate that maintenance of resting [Ca(2+)](i) in PASMCs from chronically hypoxic rats does not require activation of L-type Ca(2+) channels and suggest that ET-1-induced contraction occurs by a mechanism primarily independent of changes in [Ca(2+)](i). PMID- 11053025 TI - SB 239063, a p38 MAPK inhibitor, reduces neutrophilia, inflammatory cytokines, MMP-9, and fibrosis in lung. AB - The effects of a second generation p38 mitogen-activated protein kinase (MAPK) inhibitor, SB 239063 [trans-1-(4-hydroxycyclohexyl)-4-(4-fluorophenyl)-5-(2 methoxypyridim idi n-4-yl)imidazole; IC(50) = 44 nM vs. p38 alpha], were assessed in models that represent different pathological aspects of chronic obstructive pulmonary disease (COPD) [airway neutrophilia, enhanced cytokine formation and increased matrix metalloproteinase (MMP)-9 activity] and in a model of lung fibrosis. Airway neutrophil infiltration and interleukin (IL)-6 levels, assessed by bronchoalveolar lavage 48 h after lipopolysaccharide (LPS) inhalation, were inhibited dose dependently by 3-30 mg/kg of SB 239063 given orally twice a day. In addition, SB 239063 (30 mg/kg orally) attenuated IL-6 bronchoalveolar lavage fluid concentrations (>90% inhibition) and MMP-9 activity (64% inhibition) assessed 6 h after LPS exposure. In guinea pig cultured alveolar macrophages, SB 239063 inhibited LPS-induced IL-6 production (IC(50) of 362 nM). In a bleomycin induced pulmonary fibrosis model in rats, treatment with SB 239063 (2.4 or 4.8 mg/day via osmotic pump) significantly inhibited bleomycin-induced right ventricular hypertrophy (indicative of secondary pulmonary hypertension) and increases in lung hydroxyproline synthesis (indicative of collagen synthesis and fibrosis). Therefore, SB 239063 demonstrates activity against a range of sequelae commonly associated with COPD and fibrosis, supporting the therapeutic potential of p38 MAPK inhibitors such as SB 239063 in chronic airway disease. PMID- 11053026 TI - Inhibition of K(Ca) channels restores blunted hypoxic pulmonary vasoconstriction in rats with cirrhosis. AB - Rats with liver cirrhosis exhibit the hepatopulmonary syndrome composed of blunted hypoxic pulmonary vasoconstriction and arterial hypoxemia. The purpose of this study was to investigate the roles of nitric oxide (NO) and endothelin-1 (ET 1) in the blunted hypoxic pressor response (HPR) in rats with common bile duct ligation (CBDL). Lungs from CBDL rats exhibited markedly blunted HPR, increased endothelial NO synthase (NOS) protein expression, and decreased ET-1 mRNA and peptide expression. The blunted HPR was not reversed by sequential NOS and soluble guanylyl cyclase inhibition by nitro-L-arginine and 1H [1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ), respectively, or by NOS inhibition combined with ET-1 addition. The blunted HPR was not due to a generalized inability to vasoconstrict because perfusion pressure was equally elevated by increased perfusate KCl in CBDL and sham lungs. After KCl vasoconstriction, HPR was potentiated and did not differ between CBDL and sham lungs. Blunted HPR was also completely restored in CBDL lungs treated with nitro L-arginine, ODQ, and the Ca(2+)-activated K(+) channel blockers apamin and charybdotoxin. These results indicate that although CBDL-induced liver cirrhosis is associated with increased NO and decreased ET-1 in the lung, the blunted HPR is a result of additional factors and appears to involve Ca(2+)-activated K(+) channel activation. PMID- 11053027 TI - Selectivity properties of a Na-dependent amino acid cotransport system in adult alveolar epithelial cells. AB - We investigated the amino acid specificity of a Na-dependent amino acid cotransport system that contributes to transepithelial Na absorption in the apical membrane of cultured adult rat alveolar epithelial cell monolayers. Short circuit current was increased by basic, uncharged polar, and nonpolar amino acids but not by L-aspartic acid or L-proline. EC(50) values for L-lysine and L histidine were 0.16 and 0.058 mM, respectively. The L-lysine-stimulated short circuit current was Na dependent, with a concentration causing a half-maximal stimulation by Na of 44.24 mM. L-Serine, L-glutamine, and L-cysteine had EC(50) values of 0.095, 0.25, and 0.12 mM, respectively. L-Alanine had the highest affinity, with an EC(50) of 0.027 mM. We conclude that monolayer cultures of adult rat alveolar epithelial cells possess a broad-specificity Na-dependent amino acid cotransport system with properties consistent with system B(0,+). We suggest that this cotransport system plays a critical role in recycling of constituent amino acids that make up glutathione, thus ensuring efficient replenishment of this important antioxidant within the alveolar fluid. PMID- 11053028 TI - Vitronectin adsorption to chrysotile asbestos increases fiber phagocytosis and toxicity for mesothelial cells. AB - Biological modification of asbestos fibers can alter their interaction with target cells. We have shown that vitronectin (VN), a major adhesive protein in serum, adsorbs to crocidolite asbestos and increases fiber phagocytosis by mesothelial cells via integrins. Because chrysotile asbestos differs significantly from crocidolite in charge and shape, we asked whether VN would also adsorb to chrysotile asbestos and increase its toxicity for mesothelial cells. We found that VN, either from purified solutions or from serum, adsorbed to chrysotile but at a lower amount per surface area than to crocidolite. Nevertheless, VN coating increased the phagocytosis of chrysotile as well as of crocidolite asbestos. VN coating of both chrysotile and crocidolite, but not of glass beads, increased intracellular oxidation and apoptosis of mesothelial cells. The additional apoptosis could be blocked by integrin-ligand blockade with arginine-glycine-aspartic acid peptides, confirming a role for integrins in the fiber-induced toxicity. We conclude that VN increases the phagocytosis of chrysotile as well as of crocidolite asbestos and that phagocytosis is important in fiber-induced toxicity for mesothelial cells. PMID- 11053029 TI - Phagocytosis of particulate air pollutants by human alveolar macrophages stimulates the bone marrow. AB - Epidemiologic studies have shown an association between the level of ambient particulate matter < 10 microm (PM(10)) and cardiopulmonary mortality. We have shown that exposure of rabbits to PM(10) stimulates the bone marrow. In this study, we determined whether human alveolar macrophages (AMs) that phagocytose atmospheric PM(10) produce mediators capable of stimulating the bone marrow. AMs incubated with PM(10) for 24 h produced tumor necrosis factor-alpha in a dose dependent manner (86.8 +/- 53.29 pg/ml with medium alone; 1,087.2 +/- 257.3 pg/ml with 0.1 mg/ml of PM(10); P < 0.02). Instillation of the supernatants from AMs incubated with 0.1 mg/ml of PM(10) into the lungs of rabbits (n = 6) increased circulating polymorphonuclear leukocyte (PMN) and band cell counts as well as shortened the PMN transit time through the bone marrow (87.9 +/- 3.3 h) compared with unstimulated human AMs (104.9 +/- 2.4 h; P < 0.01; n = 5 rabbits). The supernatants from rabbit AMs incubated with 0.1 mg/ml of PM(10) (n = 4 rabbits) caused a similar shortening in the PMN transit time through the bone marrow (91.5 +/- 1.6 h) compared with human AMs. We conclude that mediators released from AMs after phagocytosis of PM(10) induce a systemic inflammatory response that includes stimulation of the bone marrow. PMID- 11053030 TI - p38 MAP kinase regulates IL-1 beta responses in cultured airway smooth muscle cells. AB - We have previously reported that interleukin (IL)-1 beta causes beta-adrenergic hyporesponsiveness in cultured human airway smooth muscle (HASM) cells by increasing cyclooxygenase (COX)-2 expression. The purpose of this study was to determine whether p38 mitogen-activated protein (MAP) kinase is involved in these events. IL-1 beta (2 ng/ml for 15 min) increased p38 phosphorylation fourfold. The p38 inhibitor SB-203580 (3 microM) decreased IL-1 beta-induced COX-2 by 70 +/ 7% (P < 0.01). SB-203580 had no effect on PGE(2) release in control cells but caused a significant (70-80%) reduction in PGE(2) release in IL-1 beta-treated cells. IL-1 beta increased the binding of nuclear proteins to the oligonucleotides encoding the consensus sequences for activator protein (AP)-1 and nuclear factor (NF)-kappa B, but SB-203580 did not affect this binding, suggesting that the mechanism of action of p38 was not through AP-1 or NF-kappa B activation. The NF-kappa B inhibitor MG-132 did not alter IL-1 beta-induced COX-2 expression, indicating that NF-kappa B activation is not required for IL-1 beta induced COX-2 expression in HASM cells. IL-1 beta attenuated isoproterenol induced decreases in HASM stiffness as measured by magnetic twisting cytometry, and SB-203580 abolished this effect. These results are consistent with the hypothesis that p38 is involved in the signal transduction pathway through which IL-1 beta induces COX-2 expression, PGE(2) release, and beta-adrenergic hyporesponsiveness. PMID- 11053031 TI - PKC, p42/p44 MAPK, and p38 MAPK are required for HGF-induced proliferation of H441 cells. AB - In this paper, we studied the signaling pathway used by hepatocyte growth factor/scatter factor (HGF) to stimulate mitosis. We show, using H441 cells, that 1) HGF activates membrane-associated protein kinase C (PKC); the activity is transient and peaks within 30 min; 2) HGF activates p42/p44 and p38 mitogen activated protein kinases (MAPKs); maximum activity in both is within 10 min; and 3) the activation of neither p38 nor p42/p44 MAPK is dependent on PKC, indicating that HGF uses separate and nonintersecting pathways to activate these two classes of kinase. However, phorbol 12-myristate 13-acetate also activates both MAPKs as well as PKC, but this activation is abolished in cells pretreated with the PKC inhibitor GF-109203X. HGF was found to significantly increase [(3)H]thymidine incorporation within 5 h; peak thymidine incorporation was observed at 16 h. However, when cells were pretreated with inhibitors of p42/p44 (PD-98059), p38 (SB-203580), or PKC (GF-109203X, Go-6983, or myristoylated inhibitor peptide(19 27)), HGF-induced thymidine uptake was diminished in a dose-dependent manner. Taken together, these results demonstrate that HGF activates PKC and both MAPKs simultaneously through parallel pathways and that the activation of the MAPKs does not depend on PKC. However, p38 and p42/p44 MAPKs and PKC may all be essential for HGF-induced proliferation of H441 cells. PMID- 11053032 TI - Surfactant components modulate fibroblast apoptosis and type I collagen and collagenase-1 expression. AB - During lung injury, fibroblasts migrate into the alveolar spaces where they can be exposed to pulmonary surfactant. We examined the effects of Survanta and surfactant protein A (SP-A) on fibroblast growth and apoptosis and on type I collagen, collagenase-1, and tissue inhibitor of metalloproteinase (TIMP)-1 expression. Lung fibroblasts were treated with 100, 500, and 1,000 microg/ml of Survanta; 10, 50, and 100 microg/ml of SP-A; and 500 microg/ml of Survanta plus 50 microg/ml of SP-A. Growth rate was evaluated by a formazan-based chromogenic assay, apoptosis was evaluated by DNA end labeling and ELISA, and collagen, collagenase-1, and TIMP-1 were evaluated by Northern blotting. Survanta provoked fibroblast apoptosis, induced collagenase-1 expression, and decreased type I collagen affecting mRNA stability approximately 10-fold as assessed with the use of actinomycin D. Collagen synthesis and collagenase activity paralleled the gene expression results. SP-A increased collagen expression approximately 2-fold and had no effect on collagenase-1, TIMP-1, or growth rate. When fibroblasts were exposed to a combination of Survanta plus SP-A, the effects of Survanta were partially reversed. These findings suggest that surfactant lipids may protect against intraluminal fibrogenesis by inducing fibroblast apoptosis and decreasing collagen accumulation. PMID- 11053033 TI - Anti-inflammatory effects of triptolide in human bronchial epithelial cells. AB - Triptolide (PG490, 97% pure) is a diterpenoid triepoxide with potent anti inflammatory and immunosuppressive effects in transformed human bronchial epithelial cells and T cells (Qiu D, Zhao G, Aoki Y, Shi L, Uyei A, Nazarian S, Ng JC-H, and Kao PN. J Biol Chem 274: 13443-13450, 1999). Triptolide, with an IC(50) of approximately 20-50 ng/ml, inhibits normal and transformed human bronchial epithelial cell expression of interleukin (IL)-6 and IL-8 stimulated by phorbol 12-myristate 13-acetate (PMA), tumor necrosis factor-alpha, or IL-1 beta. Nuclear runoff and luciferase reporter gene assays demonstrate that triptolide inhibits IL-8 transcription. Triptolide also inhibits the transcriptional activation, but not the DNA binding, of nuclear factor-kappa B. A cDNA array and clustering algorithm analysis reveals that triptolide inhibits expression of the PMA-induced genes tumor necrosis factor-alpha, IL-8, macrophage inflammatory protein-2 alpha, intercellular adhesion molecule-1, integrin beta(6), vascular endothelial growth factor, granulocyte-macrophage colony-stimulating factor, GATA 3, fra-1, and NF45. Triptolide also inhibits constitutively expressed cell cycle regulators and survival genes cyclins D1, B1, and A1, cdc-25, bcl-x, and c-jun. Thus anti-inflammatory, antiproliferative, and proapoptotic properties of triptolide are associated with inhibition of nuclear factor-kappa B signaling and inhibition of genes known to regulate cell cycle progression and survival. PMID- 11053034 TI - Fas ligand expression coincides with alveolar cell apoptosis in late-gestation fetal lung development. AB - Apoptosis plays a central role in the cellular remodeling of the developing lung. We determined the spatiotemporal patterns of the cell death regulators Fas and Fas ligand (FasL) during rabbit lung development and correlated their expression with pulmonary and type II cell apoptosis. Fetal rabbit lungs (25-31 days gestation) were assayed for apoptotic activity by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) and DNA size analysis. Fas and FasL expression were analyzed by RT-PCR, immunoblot, and immunohistochemistry. Type II cell apoptosis increased significantly on gestational day 28; the type II cell apoptotic index increased from 0.54 +/- 0.34% on gestational day 27 to 3.34 +/- 1.24% on day 28, P < 0.01 (ANOVA). This corresponded with the transition from the canalicular to the terminal sac stage of development. The day 28 rise in epithelial apoptosis was synchronous with a robust if transient 20-fold increase in FasL mRNA and a threefold increase in FasL protein levels. In contrast, Fas mRNA levels remained constant, suggestive of constitutive expression. Fas and FasL proteins were immunolocalized to alveolar type II cells and bronchiolar Clara cells. The correlation of this highly specific pattern of FasL expression with alveolar epithelial apoptosis and remodeling implicates the Fas/FasL system as a potentially important regulatory pathway in the control of postcanalicular alveolar cytodifferentiation. PMID- 11053035 TI - Secretion of extracellular superoxide dismutase in neonatal lungs. AB - Extracellular superoxide dismutase (EC-SOD), the only known enzymatic scavenger of extracellular superoxide, may modulate reactions of nitric oxide (NO) in the lungs by preventing reactions between superoxide and NO. The regulation of EC-SOD has not been examined in developing lungs. We hypothesize that EC-SOD plays a pivotal role in the response to increased oxygen tension and NO in the neonatal lung. This study characterizes rabbit EC-SOD and investigates the developmental regulation of EC-SOD activity, protein expression, and localization. Purified rabbit EC-SOD was found to have several unique biochemical attributes distinct from EC-SOD in other species. Rabbit lung EC-SOD contains predominantly uncleaved subunits that do not form disulfide-linked dimers. The lack of intersubunit disulfide bonds may contribute to the decreased heparin affinity and lower EC-SOD content in rabbit lung. EC-SOD activity in rabbit lungs is low before birth and increases soon after gestation. In addition, the enzyme is localized intracellularly in preterm and term rabbit lungs. Secretion of active EC-SOD into the extracellular compartment increases with age. The changes in EC-SOD localization and activity have implications for the neonatal pulmonary response to oxidative stress and the biological activity of NO at birth. PMID- 11053036 TI - Adoptive transfer of acute lung injury. AB - In this study, we describe a novel adoptive transfer protocol to study acute lung injury in the rat. We show that bronchoalveolar lavage (BAL) cells isolated from rats 5 h after intratracheal administration of lipopolysaccharide (LPS) induce a lung injury when transferred to normal control recipient rats. This lung injury is characterized by increased alveolar-arterial oxygen difference and extravasation of Evans blue dye (EBD) into lungs of recipient rats. Recipient rats receiving similar numbers of donor cells isolated from healthy rats do not show adverse changes in the alveolar-arterial oxygen difference or in extravasation of EBD. The adoptive transfer-induced lung injury is associated with increased numbers of neutrophils in the BAL, the levels of which are similar to the numbers observed in BAL cells isolated from rats treated for 5 h with LPS. As an indicator of BAL cell activation, donor BAL cell inducible nitric oxide synthase (iNOS) expression was compared with BAL cell iNOS expression 48 h after adoptive transfer. BAL cells isolated 5 h after LPS administration expressed iNOS immediately after isolation. In contrast, BAL cells isolated 48 h after adoptive transfer did not express iNOS immediately after isolation but expressed iNOS following a 24-h ex vivo culture. These findings indicate that the activation state of donor BAL cells differs from BAL cells isolated 48 h after adoptive transfer, suggesting that donor BAL cells may stimulate migration of new inflammatory cells into the recipient rats lungs. PMID- 11053037 TI - Hypercapnia induces injury to alveolar epithelial cells via a nitric oxide dependent pathway. AB - Ventilator strategies allowing for increases in carbon dioxide (CO(2)) tensions (hypercapnia) are being emphasized to ameliorate the consequences of inflammatory mediated lung injury. Inflammatory responses lead to the generation of reactive species including superoxide (O(2)(-)), nitric oxide (.NO), and their product peroxynitrite (ONOO(-)). The reaction of CO(2) and ONOO(-) can yield the nitrosoperoxocarbonate adduct ONOOCO(2)(-), a more potent nitrating species than ONOO(-). Based on these premises, monolayers of fetal rat alveolar epithelial cells were utilized to investigate whether hypercapnia would modify pathways of.NO production and reactivity that impact pulmonary metabolism and function. Stimulated cells exposed to 15% CO(2) (hypercapnia) revealed a significant increase in.NO production and nitric oxide synthase (NOS) activity. Cell 3 nitrotyrosine content as measured by both HPLC and immunofluorescence staining also increased when exposed to these same conditions. Hypercapnia significantly enhanced cell injury as evidenced by impairment of monolayer barrier function and increased induction of apoptosis. These results were attenuated by the NOS inhibitor N-monomethyl-L-arginine. Our studies reveal that hypercapnia modifies.NO-dependent pathways to amplify cell injury. These results affirm the underlying role of.NO in tissue inflammatory reactions and reveal the impact of hypercapnia on inflammatory reactions and its potential detrimental influences. PMID- 11053038 TI - Molecular and functional properties of two-pore-domain potassium channels. AB - The two-pore-domain K(+) channels, or K(2P) channels, constitute a novel class of K(+) channel subunits. They have four transmembrane segments and are active as dimers. The tissue distribution of these channels is widespread, and they are found in both excitable and nonexcitable cells. K(2P) channels produce currents with unusual characteristics. They are quasi-instantaneous and noninactivating, and they are active at all membrane potentials and insensitive to the classic K(+) channel blockers. These properties designate them as background K(+) channels. They are expected to play a major role in setting the resting membrane potential in many cell types. Another salient feature of K(2P) channels is the diversity of their regulatory mechanisms. The weak inward rectifiers TWIK-1 and TWIK-2 are stimulated by activators of protein kinase C and decreased by internal acidification, the baseline TWIK-related acid-sensitive K(+) (TASK)-1 and TASK-2 channels are sensitive to external pH changes in a narrow range near physiological pH, and the TWIK-related (TREK)-1 and TWIK-related arachidonic acid stimulated K(+) (TRAAK) channels are the first cloned polyunsaturated fatty acids activated and mechanogated K(+) channels. The recent demonstration that TASK-1 and TREK-1 channels are activated by inhalational general anesthetics, and that TRAAK is activated by the neuroprotective agent riluzole, indicates that this novel class of K(+) channels is an interesting target for new therapeutic developments. PMID- 11053039 TI - In vivo role of CLC chloride channels in the kidney. AB - Chloride channels in the kidney are involved in important physiological functions such as cell volume regulation, acidification of intracellular vesicles, and transepithelial chloride transport. Among eight mammalian CLC chloride channels expressed in the kidney, three (CLC-K1, CLC-K2, and CLC-5) were identified to be related to kidney diseases in humans or mice. CLC-K1 mediates a transepithelial chloride transport in the thin ascending limb of Henle's loop and is essential for urinary concentrating mechanisms. CLC-K2 is a basolateral chloride channel in distal nephron segments and is necessary for chloride reabsorption. CLC-5 is a chloride channel in intracellular vesicles of proximal tubules and is involved in endocytosis. This review will cover the recent advances in research on the CLC chloride channels of the kidney with a special focus on the issues most necessary to understand their physiological roles in vivo, i.e., their intrarenal and cellular localization and their phenotypes of humans and mice that have their loss-of-function mutations. PMID- 11053040 TI - A(2A) adenosine receptor-mediated inhibition of renal injury and neutrophil adhesion. AB - We sought to determine the mechanisms responsible for the reduced renal tissue injury by agonists of A(2A) adenosine receptors (A(2A)-ARs) in models of ischemia reperfusion (I/R) injury. DWH-146e, a selective A(2A)-AR agonist, was administered subcutaneously to Sprague-Dawley rats and C57BL/6 mice via osmotic minipumps, and animals were subjected to I/R. I/R led to an increase in plasma creatinine and kidney neutrophil infiltration. Infusion of DWH-146e at 10 ng. kg( 1). min(-1) produced a 70% reduction in plasma creatinine as well as a decrease in neutrophil density in outer medulla and cortex and myeloperoxidase activity in the reperfused kidney. Myeloperoxidase activity in kidney correlated with the degree of renal injury. P-selectin and intercellular adhesion molecule 1 (ICAM-1) immunoreactivity were most prominent in endothelial cells of peritubular capillaries and interlobular arteries of cortex and outer and inner medulla of vehicle-treated mice whose kidneys were subjected to I/R. DWH-146e treatment led to a pronounced decrease in P-selectin- and ICAM-1-like immunoreactivity. These data are consistent with our hypothesis that A(2A)-AR agonists limit I/R injury due to an inhibitory effect on neutrophil adhesion. PMID- 11053041 TI - Renin expression in COX-2-knockout mice on normal or low-salt diets. AB - Experiments were performed in mice to investigate whether cyclooxygenase-2 (COX 2) in epithelial cells near the tubulovascular contact point (macula densa and TAL cells) may regulate renin gene expression in juxtaglomerular granular cells. Renin activity, afferent arteriolar granularity, and renin mRNA were determined in wild-type mice and in COX-2-knockout mice on control and low-NaCl diets. Renin activity in microdissected glomeruli assessed as angiotensin I formation in the presence of excess substrate and afferent arteriolar granularity determined by direct visualization and immunostaining were significantly reduced in COX-2 -/- compared with wild-type animals. Similarly, renal cortical mRNA levels were lower in COX-2 -/- than in wild-type mice. Maintaining mice on a low-salt diet for 14 days induced an increase in renin mRNA, afferent arteriolar granularity, and renin activity in wild-type mice. In contrast, renin mRNA and renin granularity did not significantly increase in low-salt-treated COX-2 -/- mice, whereas the increase in juxtaglomerular renin enzyme activity was markedly attenuated, but not fully blocked. In additional experiments we found that COX-2 mRNA was increased in angiotensin type 1A receptor-knockout mice compared with wild-type mice. We conclude that COX-2 in the tubulovascular contact region is a critical determinant of renin synthesis in granular cells under resting conditions and that it participates in the stimulation of renin expression caused by a low-NaCl intake. PMID- 11053042 TI - Basolateral localization of organic cation transporter 2 in intact renal proximal tubules. AB - The localization of organic cation transporter 2 (OCT2) within renal cells is the subject of considerable controversy, resulting in marked uncertainty as to its function. To resolve this issue, we made an OCT2/green fluorescent protein (GFP) fusion construct (rOCT2-GFP) and determined its localization within Xenopus laevis oocytes and renal cells using confocal microscopy. Oocytes expressing rOCT2-GFP exhibited plasma membrane fluorescence as well as greatly increased specific, potential-driven uptake of [(14)C]tetraethylammonium (TEA). Polarized monolayers of renal epithelial cell lines [LLC-PK(1) and Madin-Darby canine kidney (MDCK)] transiently transfected with pEGFP-C3, which codes for a cytoplasmic GFP, showed a diffuse, evenly distributed cytoplasmic signal with no plasma membrane fluorescence. In contrast, cells transiently transfected with pEGFP-C3/rOCT2 (the vector coding for rOCT2-GFP) showed predominantly plasma membrane fluorescence, which was most prominent in the lateral membrane. MDCK cells stably expressing rOCT2-GFP (MDCK/rOCT2-GFP) maintained in long-term culture showed a greatly increased basal and lateral membrane fluorescence. When grown on porous supports, MDCK/rOCT2-GFP monolayers showed specific, potential driven TEA uptake from the basal side. Finally, expression and distribution of rOCT2-GFP were investigated in isolated killifish (Fundulus heteroclitus) renal proximal tubules. On expression of rOCT2-GFP, transfected tubules showed marked basal and lateral membrane fluorescence, with no detectable signal at the apical membrane. In contrast, tubules expressing a luminal sodium-dicarboxylate cotransporter (rbNaDC-1)-GFP construct showed apical membrane fluorescence, and tubules expressing cytoplasmic GFP had a diffuse cytoplasmic fluorescence. These results indicate that rOCT2 is basolateral in renal proximal tubule cells. PMID- 11053043 TI - Angiotensin II increases vasopressin-stimulated facilitated urea permeability in rat terminal IMCDs. AB - Angiotensin II receptors are present along the rat inner medullary collecting duct (IMCD), although their physiological role is unknown. Because urea is one of the major solutes transported across the terminal IMCD, we measured angiotensin II's effect on urea permeability. In the perfused rat terminal IMCD, angiotensin II had no effect on basal urea permeability but significantly increased vasopressin-stimulated urea permeability by 55%. Angiotensin II, both without and with vasopressin, also increased the amount of (32)P incorporated into urea transporter (UT)-A1 in inner medullary tissue exposed to these hormones ex vivo. Because angiotensin II activates protein kinase C, we tested the effect of staurosporine (SSP). In the absence of angiotensin II, SSP had no effect on vasopressin-stimulated urea permeability in the perfused terminal IMCD. However, SSP completely and reversibly blocked the angiotensin II-mediated increase in vasopressin-stimulated urea permeability. SSP and chelerythrine reduced the angiotensin II-stimulated (32)P incorporation into UT-A1 in inner medullary tissue exposed ex vivo. We conclude that angiotensin II increases vasopressin stimulated facilitated urea permeability and (32)P incorporation into the 97- and 117-kDa UT-A1 proteins via a protein kinase C-mediated signaling pathway. These data suggest that angiotensin II augments vasopressin-stimulated facilitated urea transport in the rat terminal IMCD and may play a physiological role in the urinary concentrating mechanism by augmenting the maximal response to vasopressin. PMID- 11053044 TI - Pathways for angiotensin-(1---7) metabolism in pulmonary and renal tissues. AB - Two of the primary sites of actions for angiotensin (ANG)-(1---7) are the vasculature and the kidney. Because little information exists concerning the metabolism of ANG-(1---7) in these tissues, we investigated the hydrolysis of the peptide in rat lung and renal brush-border membrane (BBM) preparations. Radiolabeled ANG-(1---7) was hydrolyzed primarily to ANG-(1---5) by pulmonary membranes. The ANG-converting enzyme (ACE) inhibitor lisinopril abolished the generation of ANG-(1---5), as well as that of smaller metabolites. Kinetic studies of the hydrolysis of ANG-(1---7) to ANG-(1---5) by somatic (pulmonary) and germinal (testes) forms of rat ACE yielded similar values, suggesting that the COOH-domain is responsible for the hydrolysis of ANG-(1---7). Pulmonary metabolism of ANG-(1---5) yielded ANG-(3---5) and was independent of ACE but may involve peptidyl or dipeptidyl aminopeptidases. In renal cortex BBM, ANG-(1---7) was rapidly hydrolyzed to mono- and dipeptide fragments and ANG-(1---4). Aminopeptidase (AP) inhibition attenuated the hydrolysis of ANG-(1---7) and increased ANG-(1---4) formation. Combined treatment with AP and neprilysin (Nep) inhibitors abolished ANG-(1---4) formation and preserved ANG-(1---7). ACE inhibition had no effect on the rate of hydrolysis or the metabolites formed in the BBM. In conclusion, ACE was the major enzymatic activity responsible for the metabolism of ANG-(1---7) in the lung, which is consistent with the ability of ACE inhibitors to increase the half-life of circulating ANG-(1---7) and raise endogenous levels of the peptide. An alternate pathway of metabolism was revealed in the renal cortex, where increased AP and Nep activities, relative to ACE activity, promote conversion of ANG-(1---7) to ANG-(1---4) and smaller fragments. PMID- 11053045 TI - Functional expression of novel peptide transporter in renal basolateral membranes. AB - We examined the peptide transport activity in renal basolateral membranes. [(14)C]glycylsarcosine (Gly-Sar) uptake in rat renal cortical slices was saturable and inhibited by excess dipeptide and aminocephalosporin cefadroxil. When several renal cell lines were screened for the basolateral peptide transport activity, Madin-Darby canine kidney (MDCK) cells were demonstrated to have the greatest transport activity. [(14)C]Gly-Sar uptake across the basolateral membranes of MDCK cells was inhibited by di- and tripeptide and decreased with decreases in extracellular pH from 7.4 to 5.0. The Michaelis-Menten constant value of [(14)C]Gly-Sar uptake across the basolateral membranes of MDCK cells was 71 microM. The basolateral peptide transporter in MDCK cells showed several different [(14)C]Gly-Sar transport characteristics in growth dependence, pH profile, substrate affinity, and sensitivities to chemical modifiers from those of the apical H(+)-peptide cotransporter of MDCK cells. The findings of the present investigation indicated that the peptide transporter was expressed in the renal basolateral membranes. In addition, from the functional characteristics, the renal basolateral peptide transporter was suggested to be distinguishable from known peptide transporters, i.e., H(+)-peptide cotransporters (PEPT1 and PEPT2) and the intestinal basolateral peptide transporter. PMID- 11053046 TI - Dynamic interaction between myogenic and TGF mechanisms in afferent arteriolar blood flow autoregulation. AB - The dynamic activity of afferent arteriolar diameter (AAD) and blood flow (AABF) responses to a rapid step increase in renal arterial pressure (100-148 mmHg) was examined in the kidneys of normal Sprague-Dawley rats (n = 11) before [tubuloglomerular feedback (TGF)-intact] and after interruption of distal tubular flow (TGF-independent). Utilizing the in vitro blood-perfused juxtamedullary nephron preparation, fluctuations in AAD and erythrocyte velocity were sampled by using analog-to-digital computerized conversion, video microscopy, image shearing, and fast-frame, slow-frame techniques. These assessments enabled dynamic characterization of the autonomous actions and collective interactions between the myogenic and TGF mechanisms at the level of the afferent arteriole. The TGF-intact and TGF-independent systems exhibited common initial (0-24 vs. 0 13 s, respectively) response slope kinetics (-0.53 vs. -0.47% DeltaAAD/s; respectively) yet different maximum vasoconstrictive magnitude (-11.28 +/- 0.1 vs. -7. 02 +/- 0.9% DeltaAAD; P < 0.05, respectively). The initial AABF responses similarly exhibited similar kinetics but differing magnitudes. In contrast, during the sustained pressure input (13-97 s), the maximum vasoconstrictor magnitude (-7.02 +/- 0.9% DeltaAAD) and kinetics (-0.01% DeltaAAD/s) of the TGF independent system were markedly blunted whereas the TGF-intact system exhibited continued vasoconstriction with slower kinetics (-0.20% DeltaAAD/s) until a steady-state plateau was reached (-25.9 +/- 0.4% DeltaAAD). Thus the TGF mechanism plays a role in both direct mediation of vasoconstriction and in modulation of the myogenic response. PMID- 11053047 TI - Mapping and functional analysis of an instability element in phosphoenolpyruvate carboxykinase mRNA. AB - Phosphoenolpyruvate carboxykinase (PEPCK) is a key regulatory enzyme of renal gluconeogenesis. The 3'-nontranslated region of the PEPCK mRNA contains an instability element that facilitates its rapid turnover and contributes to the regulation of PEPCK gene expression. Such processes are mediated by specific protein-binding elements. Thus RNA gel shift analysis was used to identify proteins in rat renal cortical cytosolic extracts that bind to the 3' nontranslated region of the PEPCK mRNA. Deletion constructs were then used to map the binding interactions to two adjacent RNA segments (PEPCK-6 and PEPCK-7). However, competition experiments established that only the binding to PEPCK-7 was specific. Functional studies were performed by cloning similar segments in a luciferase reporter construct, pLuc/Zeo. This analysis indicated that both PEPCK 6 and PEPCK-7 segments were necessary to produce a decrease in luciferase activity equivalent to that observed with the full-length 3'-nontranslated region. Thus the PEPCK-7 segment binds a specific protein that may recruit one or more proteins to form a complex that mediates the rapid decay of the PEPCK mRNA. PMID- 11053048 TI - Expression of aquaporins in the renal connecting tubule. AB - The renal connecting tubule (CNT) is a distinct segment that occurs between the distal convoluted tubule (DCT) and the cortical collecting duct. On the basis of its characterization in rabbit it is widely believed that connecting tubule cells have a low permeability to water and do not respond to vasopressin. Here we utilize segment-specific markers and specific aquaporin antibodies to characterize expression of water channels in CNT of the rat by immunocytochemistry. Colocalization of aquaporin 2 (AQP2), AQP3, and AQP4 with Na(+), Ca(2+) exchanger (NCX), a transporter characteristic of the connecting tubule, gave heterogeneous labeling. There was aquaporin labeling in many but not all regions labeled by NCX. Colocalization of AQP2 with AQP3 and with AQP4 showed that AQP3 and AQP4 labeling were always accompanied by AQP2. Immunogold labeling and electron microscopy showed that NCX-labeled cells with AQP2 labeling had the morphology of CNT cells, whereas NCX-labeled cells without AQP2 labeling were DCT cells. The latter regions were identified as the late region of the DCT known as DCT2. Additionally, regions of CNT lacking AQP2 labeling could be identified in Brattleboro rats not treated with vasopressin but not in such animals chronically treated with deamino-Cys(1),D-Arg(8)-vasopressin (dDAVP). Quantitative analysis of labeling was consistent with expression of AQP2 over a longer region of CNT after dDAVP exposure. PMID- 11053049 TI - Gentamicin traffics rapidly and directly to the Golgi complex in LLC-PK(1) cells. AB - To study the intracellular mechanisms of aminoglycoside toxicity, we used a 1:1 fluorescent conjugate of Texas Red and gentamicin (TRG) to quantify early uptake dynamics in renal epithelial (LLC-PK(1)) cells. Utilizing a protocol that quenches TRG fluorescence from lysosomes, the bulk of intracellular accumulation, we determined a portion rapidly trafficked directly to the Golgi complex when identified by a FITC-conjugated lectin from Lens culinaris agglutinin (LCA). A kinetic study over 120 min on cells showing total and quenched TRG fluorescence was then carried out, and the fluorescence intensity from the images was quantified. Trafficking of TRG to the Golgi complex occurred within 15 min and accounted for approximately 20% of total cellular accumulation in the kinetic study. Colocalization studies using compartment-specific markers, 6-[N-(7 nitrobenz-2-oxa-1,3-diazol-4-yl)amino]hexanoyl sphingosine (C6-NBD ceramide) and LCA, for the TGN trans-Golgi network, and the cis/medial-Golgi compartments, respectively, determined colocalization occurred with both Golgi compartments. These data support the existence of a pathway that directly and rapidly shuttles a portion of internalized gentamicin to the Golgi complex. We believe this pathway may be responsible for the early negative effects seen on protein synthesis in renal proximal epithelia after aminoglycoside administration. PMID- 11053050 TI - Matrix metalloproteinases and their inhibitors regulate in vitro ureteric bud branching morphogenesis. AB - Mammalian kidney development is initiated by the mutual interaction between embryonic metanephric mesenchyme (MM) and the ureteric bud (UB), leading to tightly controlled UB branching morphogenesis. In a three-dimensional cell culture model, which employs MM cell-derived conditioned medium (BSN-CM) to induce UB cell branching morphogenesis in extracellular matrix (ECM) gels (Sakurai H, Barros EJ, Tsukamoto T, Barasch J, and Nigam SK. Proc Natl Acad Sci USA 94: 6279-6284, 1997), branching morphogenesis was inhibited by both chemical agents (ilomastat and 1,10-orthophenanthroline) and a physiological protein factor [tissue inhibitor of metalloproteinases (TIMP)-2], known to act as matrix metalloproteinase (MMP) inhibitors. In addition, UB branching was inhibited in isolated UB culture (Qiao J, Sakurai H, and Nigam SK. Proc Natl Acad Sci USA 96: 7330-7335, 1999) by TIMP-2 and ilomastat, suggesting a direct role for MMPs in UB branching. Gelatin zymography and enzymatic measurement of MMP activity revealed that MMPs could originate from at least three different sources: the conditioned medium, the ECM, and the UB cells themselves. In the UB cells, transcription of several MMPs [gelatinase A (MMP2) and B (MMP9), stromelysin (MMP3), MT1-MMP] and TIMPs was altered by BSN-CM and changed as more complex branching structures formed. The ECM appeared to serve as both a reservoir for MMPs and modulated their expression because different ECM compositions altered the total MMP activity as well as specific subsets of MMPs expressed by the UB cells (as determined by zymography and Northern analysis). In the context of UB branching morphogenesis during kidney development, our data suggest a complex model in which soluble factors produced by the MM, in the context of specific ECM components, modulate the expression of specific subsets of MMPs and TIMPs in the UB, which alter as structures develop and the matrix environment changes. This suggests distinct roles for different subsets of MMPs and their inhibitors during different phases of branching morphogenesis. PMID- 11053051 TI - Immunolocalization of electroneutral Na-HCO(3)(-) cotransporter in rat kidney. AB - An electroneutral Na-HCO(3)(-) cotransporter (NBC(N)1) was recently cloned, and Northern blot analyses indicated its expression in rat kidney. In this study, we determined the cellular and subcellular localization of NBC(N)1 in the rat kidney at the light and electron microscopic level. A peptide-derived antibody was raised against the COOH-terminal amino acids of NBC(N)1. The affinity-purified antibody specifically recognized one band, approximately 180 kDa, in rat kidney membranes. Peptide-N-glycosidase F deglycosylation reduced the band to approximately 140 kDa. Immunoblotting of membrane fractions from different kidney regions demonstrated strong signals in the inner stripe of the outer medulla (ISOM), weaker signals in the outer stripe of the outer medulla and inner medulla, and no labeling in cortex. Immunocytochemistry demonstrated that NBC(N)1 immunolabeling was exclusively observed in the basolateral domains of thick ascending limb (TAL) cells in the outer medulla (strongest in ISOM) but not in the cortex. In addition, collecting duct intercalated cells in the ISOM and in the inner medulla also exhibited NBC(N)1 immunolabeling. Immunoelectron microscopy demonstrated that NBC(N)1 labeling was confined to the basolateral plasma membranes of TAL and collecting duct type A intercalated cells. Immunolabeling controls were negative. By using 2, 7-bis-carboxyethyl-5,6 caboxyfluorescein, intracellular pH transients were measured in kidney slices from ISOM and from mid-inner medulla. The results revealed DIDS-sensitive, Na- and HCO(3)(-)-dependent net acid extrusion only in the ISOM but not in mid-inner medulla, which is consistent with the immunolocalization of NBC(N)1. The localization of NBC(N)1 in medullary TAL cells and medullary collecting duct intercalated cells suggests that NBC(N)1 may be important for electroneutral basolateral HCO(3)(-) transport in these cells. PMID- 11053052 TI - Melatonin attenuates acute renal failure and oxidative stress induced by mercuric chloride in rats. AB - We evaluated the effect of melatonin (Mel), a potent scavenger of reactive oxygen species, in the course of HgCl(2)-induced acute renal failure. Rats received by gastric gavage 1 mg/kg of Mel (n = 21) or vehicle (n = 21), 30 min before the subcutaneous injection of HgCl(2) (2.5 mg/kg). Rats were killed at 24, 48, and 72 h, and plasma creatinine (S(cr)), renal histology, proliferative activity, apoptosis, and superoxide-producing cells were studied. We also determined the renal content of malondialdehyde (MDA) and glutathione (GSH) and the activities of glutathione peroxidase and catalase. Mel pretreatment (Mel plasma levels of 3.40 +/- 3.15 microgram/ml at the time of HgCl(2) injection) prevented the increment in S(cr) and reduced tubular necrosis from 41.0 +/- 10.5 to 4.2 +/- 5.1% of proximal tubules (P < 0.01). Apoptosis and postnecrotic proliferative activity were twice more intense in the group untreated with Mel. Increment in renal content of MDA and decrease in GSH resulting from HgCl(2) toxicity were prevented by Mel. Mel also induced an important reduction in superoxide-positive cells. In contrast to the beneficial effects of pretreatment with Mel, the administration of Mel in conjunction with HgCl(2) had no effect on the oxidative damage and did not prevent nephrotoxicity. We conclude that the beneficial effects of pharmacological doses of Mel are due to its antioxidant properties. PMID- 11053053 TI - PKA site mutations of ROMK2 channels shift the pH dependence to more alkaline values. AB - Close similarity between the rat native low-conductance K(+) channel in the apical membrane of renal cortical collecting duct principal cells and the cloned rat ROMK channel strongly suggest that the two are identical. Prominent features of ROMK regulation are a steep pH dependence and activation by protein kinase A (PKA)-dependent phosphorylation. In this study, we investigated the pH dependence of cloned renal K(+) channel (ROMK2), wild-type (R2-WT), and PKA site mutant channels (R2-S25A, R2-S200A, and R2-S294A). Ba(2+)-sensitive outward whole cell currents (holding voltage -50 mV) were measured in two-electrode voltage-clamp experiments in Xenopus laevis oocytes expressing either R2-WT or mutant channels. Intracellular pH (pH(i)) was measured with pH-sensitive microelectrodes in a different group of oocytes from the same batch on the same day. Resting pH(i) of R2-WT and PKA site mutants was the same: 7.32 +/- 0.02 (n = 22). The oocytes were acidified by adding 3 mM Na butyrate with external pH (pH(o)) adjusted to 7.4, 6.9, 6.4, or 5.4. At pH(o) 7.4, butyrate led to a rapid (tau: 163 +/- 14 s, where tau means time constant, n = 4) and stable acidification of the oocytes (DeltapH(i) 0.13 +/- 0. 02 pH units, where Delta means change, n = 12). Intracellular acidification reversibly inhibited ROMK2-dependent whole cell current. The effective acidic dissociation constant (pK(a)) value of R2-WT was 6.92 +/- 0.03 (n = 8). Similarly, the effective pK(a) value of the N-terminal PKA site mutant R2-S25A was 6.99 +/- 0.02 (n = 6). The effective pK(a) values of the two COOH-terminal PKA site mutant channels, however, were significantly shifted to alkaline values; i.e., 7.15 +/- 0.06 (n = 5) for R2-S200A and 7.16 +/- 0.03 (n = 8) for R2-S294A. The apparent DeltapH shift between the R2-WT and the R2-S294A mutant was 0.24 pH units. In excised inside-out patches, alkaline pH 8.5 activated R2-S294A channel current by 32 +/- 6.7%, whereas in R2-WT channel patches alkalinzation only marginally increased current by 6.5 +/- 1% (n = 5). These results suggest that channel phosphorylation may substantially influence the pH sensitivity of ROMK2 channel. Our data are consistent with the hypothesis that in the native channel PKA activation involves a shift of the pK(a) value of ROMK channels to more acidic values, thus relieving a H(+)-mediated inhibition of ROMK channels. PMID- 11053055 TI - Interaction of angiotensin II and atrial natriuretic peptide on pH(i) regulation in MDCK cells. AB - The effect of ANG II and atrial natriuretic peptide (ANP) on intracellular pH (pH(i)) and cytosolic free calcium concentration ([Ca(2+)](i)) was investigated in Madin-Darby canine kidney cells by using the fluorescent probes 2', 7'-bis(2 carboxyethyl)-5(6)-carboxyfluorescein-acetoxymethyl ester (AM) and fura 2-AM or fluo 4-AM. pH(i) recovery rate was examined in the first 2 min after the acidification of pH(i) with a NH(4)Cl pulse. In the control situation, the pH(i) recovery rate was 0.088 +/- 0.014 pH units/min (n = 14); in the absence of external Na(+), this value was decreased. ANG II (10(-12) or 10(-9) M) caused an increase in this value, but ANG II (10(-7) M) decreased it. ANP (10(-6) M) or dimethyl-1,2-bis(2-aminophenoxy)ethane-N,N,N', N'-tetraacetic acid (BAPTA)-AM (50 microM) alone did not affect this value but impaired both stimulatory and inhibitory effects of ANG II. ANG II (10(-12), 10(-9), or 10(-7) M) increased [Ca(2+)](i) progressively from 99 +/- 10 (n = 20) to 234 +/- 7 mM (n = 10). ANP or dimethyl-BAPTA-AM decreases [Ca(2+)](i), and the subsequent addition of ANG II caused a recovery of [Ca(2+)](i) but without reaching ANG II values found in the absence of these agents. The results indicate a role for [Ca(2+)](i) in regulating the process of pH(i) recovery mediated by the Na(+)/H(+) exchanger, stimulated/impaired by ANG II, and not affected by ANP or ANG II plus ANP. This hormonal interaction may represent physiologically relevant regulation in conditions of volume alterations in the intact animal. PMID- 11053054 TI - Anaerobic and aerobic pathways for salvage of proximal tubules from hypoxia induced mitochondrial injury. AB - We have further examined the mechanisms for a severe mitochondrial energetic deficit, deenergization, and impaired respiration in complex I that develop in kidney proximal tubules during hypoxia-reoxygenation, and their prevention and reversal by supplementation with alpha-ketoglutarate (alpha-KG) + aspartate. The abnormalities preceded the mitochondrial permeability transition and cytochrome c loss. Anaerobic metabolism of alpha-KG + aspartate generated ATP and maintained mitochondrial membrane potential. Other citric-acid cycle intermediates that can promote anaerobic metabolism (malate and fumarate) were also effective singly or in combination with alpha-KG. Succinate, the end product of these anaerobic pathways that can bypass complex I, was not protective when provided only during hypoxia. However, during reoxygenation, succinate also rescued the tubules, and its benefit, like that of alpha-KG + malate, persisted after the extra substrate was withdrawn. Thus proximal tubules can be salvaged from hypoxia-reoxygenation mitochondrial injury by both anaerobic metabolism of citric-acid cycle intermediates and aerobic metabolism of succinate. These results bear on the understanding of a fundamental mode of mitochondrial dysfunction during tubule injury and on strategies to prevent and reverse it. PMID- 11053056 TI - Spontaneous shift in transcriptional profile of explanted glomeruli via activation of the MAP kinase family. AB - To understand how isolation and explantation of glomeruli affect the function of resident cells, the present study investigated the transcriptional profile of explanted normal glomeruli. We found that ex vivo incubation of glomeruli spontaneously expressed monocyte chemoattractant protein-1 (MCP-1) and stromelysin, the genes regulated by activator protein-1 (AP-1). The expression was suppressed by heparin and quercetin, the drugs with anti-AP-1 activities. The gene expression was preceded by 1) induction of AP-1 components c-fos and c-jun and 2) phosphorylation of extracellular signal-regulated kinase (ERK), p38 mitogen-activated protein (MAP) kinase, and c-Jun NH(2)-terminal kinase (JNK), the upstream inducers/activators of AP-1. Suppression of ERK by PD098059 abrogated induction of c-fos and c-jun, and the p38 MAP kinase inhibitor SB203580 attenuated c-fos expression. Furthermore, treatment with either PD098059, SB203580, or the JNK-AP-1 inhibitor curcumin diminished the expression of MCP-1 and stromelysin. The transcriptional profile of glomerular cells thus alters dramatically after explantation of glomeruli. It is, at least in part, due to activation of multiple MAP kinases that lead to induction of AP-1-dependent gene expression. PMID- 11053057 TI - Method for measuring luminal efflux of fluorescent organic compounds in isolated, perfused renal tubules. AB - To examine directly in real time the efflux of organic compounds [e. g., organic anions (OAs) such as fluorescein (FL)] across the luminal membrane of isolated, perfused renal tubules during net secretion, we devised an approach utilizing a recently developed epifluorescence microscopy system for continuous monitoring of fluorescence in the collected perfusate. To illustrate this approach, we measured the luminal efflux rate of FL in mineral oil-covered, isolated, perfused S2 segments of rabbit renal proximal tubules. The washout profile of FL showed a deviation from linearity at time 0 when plotted on a semilog scale, indicating that the luminal efflux of FL was a saturable process. We were able for the first time to determine the kinetic parameters of luminal efflux [FL concentration at one-half maximal FL efflux (K(t)(lumen)) of approximately 560 microM and maximal rate of FL efflux across the luminal membrane (J(max)(lumen)) of approximately 635 fmol. min(-1). mm(-1)]. From the present study, we conclude that the transport step for OAs across the luminal membrane of OAs is a carrier-mediated process. This approach will work to measure luminal transport in real time for any secreted organic compound that is sufficiently fluorescent to be measured with commonly available, highly sensitive optical equipment. PMID- 11053059 TI - Concurrence of sarcoidosis and aortitis: case report and review of the literature. AB - Takayasu arteritis (TA) is a rare manifestation of systemic large vessel vasculitis which affects predominantly the aorta and its main branches, but often remains unrecognised owing to delayed diagnosis and non-characteristic clinical features. Sarcoidosis, too, is a systemic inflammatory disease which can affect virtually any organ system. Reports about the coincidence of both diseases have appeared. The case presented here is characterised by a significant time lag between detection of TA and appearance of clinical signs of sarcoidosis. The woman, now 39 years old, had erythema nodosum, circumscript alopecia, and recurrent uveitis, which dated back to 1980 and was attributed to sarcoidosis. At least 12 years later aortic valve insufficiency with progressive cardiac failure developed. Histology performed at the time of aortic valve prosthesis in 1997 disclosed a diagnosis of TA, which was confined to the aortic root. Incidentally, sarcoidosis was diagnosed in adjacent lymph nodes. A thorough check up failed to detect further manifestations of TA; thus, possibly, the patients had aortitis similar to, but not identical with, TA. Several related cases previously reported are discussed, suggesting that both diseases may be inherently related as they are characterised by certain non-specific, immunoinflammatory abnormalities. This case report suggests that the prevalence of TA, or related forms of arteritis, may be higher than expected and should be considered, especially in younger patients with non-characteristic cardiovascular symptoms and suspected systemic inflammatory disease. Moreover, the association with sarcoidosis in this and other previously described cases suggests that the two diseases may be related and that TA or TA-like vasculitis may even be a complication of sarcoidosis. PMID- 11053058 TI - Leflunomide: mode of action in the treatment of rheumatoid arthritis. AB - Leflunomide is a selective inhibitor of de novo pyrimidine synthesis. In phase II and III clinical trials of active rheumatoid arthritis, leflunomide was shown to improve primary and secondary outcome measures with a satisfactory safety profile. The active metabolite of leflunomide, A77 1726, at low, therapeutically applicable doses, reversibly inhibits dihydroorotate dehydrogenase (DHODH), the rate limiting step in the de novo synthesis of pyrimidines. Unlike other cells, activated lymphocytes expand their pyrimidine pool by approximately eightfold during proliferation; purine pools are increased only twofold. To meet this demand, lymphocytes must use both salvage and de novo synthesis pathways. Thus the inhibition of DHODH by A77 1726 prevents lymphocytes from accumulating sufficient pyrimidines to support DNA synthesis. At higher doses, A77 1726 inhibits tyrosine kinases responsible for early T cell and B cell signalling in the G(0)/G(1) phase of the cell cycle. Because the immunoregulatory effects of A77 1726 occur at doses that inhibit DHODH but not tyrosine kinases, the interruption of de novo pyrimidine synthesis may be the primary mode of action. Recent evidence suggests that the observed anti-inflammatory effects of A77 1726 may relate to its ability to suppress interleukin 1 and tumour necrosis factor alpha selectively over their inhibitors in T lymphocyte/monocyte contact activation. A77 1726 has also been shown to suppress the activation of nuclear factor kappaB, a potent mediator of inflammation when stimulated by inflammatory agents. Continuing research indicates that A77 1726 may downregulate the glycosylation of adhesion molecules, effectively reducing cell-cell contact activation during inflammation. PMID- 11053060 TI - Unusual and memorable. Reactive amyloidosis in patients receiving long term haemodialysis. PMID- 11053061 TI - Health impact of pain in the hip region with and without radiographic evidence of osteoarthritis: a study of new attenders to primary care. The PCR Hip Study Group. AB - OBJECTIVES: To assess the health impact of hip pain at the time of first presentation to primary care, and the influence on this of radiographic evidence of osteoarthritis. SUBJECTS AND METHODS: Cross sectional survey of 195 patients (63 male, 132 female), aged 40 years and over, presenting with a new episode of hip pain, recruited from 35 general practices across the UK. Health status at presentation was determined by a structured questionnaire on symptoms, healthcare use, and health related quality of life (SF-36). Pelvic radiographs were assessed blindly for hip osteoarthritis using standard scoring systems. RESULTS: The overall impact on health was substantial. Before their first consultation, three quarters of patients needed analgesics, half used topical creams or ointments, and one in eight used a walking stick. Most of these impact measures were, however, unrelated to the degree of radiographic change, though use of a walking stick was increased in those with the most severe damage. Health status, as judged by the SF-36, was also impaired for measures of physical function and pain, but the impact on the "mental health", "general health", and "vitality" dimensions was small. There was a weak relation between the SF-36 scores and radiographic change, with many domains unrelated to the severity of radiographic damage. CONCLUSIONS: This study is the first to show the therapeutic impact and pattern of impairment in health status resulting from hip pain at the time of first presentation to the healthcare services. Unlike many regional pain syndromes seen in primary care, such as back pain, hip pain does not impact on wider aspects of quality of life, such as general health status, mental health, or vitality. Furthermore, any impact of hip pain in this group is not markedly influenced by the degree of structural damage. Further follow up is required to determine whether such damage influences the persistence of any adverse impact. PMID- 11053062 TI - Inflammatory arthritis in children with osteochondrodysplasias. AB - Osteochondrodysplasias are a heterogeneous group of genetic skeletal dysplasias. Patients with these diseases commonly develop an early degenerative arthritis or osteoarthritis. Occasional observations of inflammatory arthritis have been made in this population but such observations are based on clinical grounds alone without confirmatory imaging studies. Four patients followed up in a paediatric rheumatology clinic with three different skeletal dysplasias, who had both clinical and radiological evidence of an inflammatory arthritis and coexistent degenerative arthritis, are described. PMID- 11053063 TI - Digital vascular responses and serum endothelin-1 concentrations in primary and secondary Raynaud's phenomenon. AB - OBJECTIVE: To determine circulating endothelin-1 levels (ET-1) in patients with primary or secondary associated Raynaud's phenomenon (RP) under resting conditions and in response to cold provocation. METHODS: Patients were categorised as primary RP (18) or scleroderma associated RP (14). Finger blood flow was measured by venous occlusion plethysmography at finger temperatures of 32 degrees C and 24 degrees C. Vasospasm was detected as a finger systolic pressure of 0 mm Hg after standardised provocative cooling. Severity of vasospasm was assessed by the level of cooling required to provoke spasm. Plasma ET-1 levels were measured in antecubital blood withdrawn under baseline conditions (finger 32 degrees C) and at the point of vasospasm. Measurements were also made in 19 matched control subjects. RESULTS: Finger blood flow was lower in patients with RP than in controls, with no difference between the two RP groups. Vasospasm occurred in all patients with RP but not in any control subjects and a grading system of severity was established. Baseline plasma ET-1 levels were similar in patients with RP and controls. Increases in ET-1 levels at the point of vasospasm in patients or corresponding timepoint in controls were also similar. There was no significant difference between the ET-1 levels in the two RP subgroups when the fingers were warm or when vasospasm was present. CONCLUSIONS: These results do not support the hypothesis that ET-1 plays a part in the pathogenesis of RP. Objective testing is a useful adjunct to the clinical diagnosis of RP and allows assignment of a severity grade. PMID- 11053064 TI - Fatigue in primary Sjogren's syndrome: is there a link with the fibromyalgia syndrome? AB - OBJECTIVE: To determine whether fibromyalgia (FM) is more common in patients with primary Sjogren's syndrome (pSS) who complain of fatigue. The association and prevalence of fatigue and FM was recorded in a group of patients with pSS and a control group of lupus patients, a subset of whom had secondary Sjogren's syndrome (sSS). METHODS: 74 patients with pSS and 216 patients with lupus were assessed with a questionnaire to identify the presence of fatigue and generalised pain. From the lupus group, in a subset of 117 lupus patients (from the Bloomsbury unit) those with sSS were identified. All patients were studied for the presence of FM. RESULTS: 50 of 74 patients with pSS (68%) reported fatigue-a prevalence significantly higher than in the lupus group (108/216 (50%); p<0.0087). Fatigue was present in 7/13 (54%) patients with SLE/sSS. FM was present in 9/74 patients with pSS (12%), compared with 11/216 lupus patients (5%), and in none of the patients with SLE/sSS. None of these values corresponds with previously reported figures of the incidence of FM in pSS. CONCLUSION: The results show that fatigue in patients with pSS and sSS is not due to the coexistence of FM in most cases. A lower incidence in the United Kingdom of FM in patients with pSS was found than has been previously reported. PMID- 11053065 TI - Comparison of the caudal and lumbar approaches to the epidural space. AB - OBJECTIVES: To investigate the accuracy of placement of epidural injections using the lumbar and caudal approaches. To identify which factors, if any, predicted successful placement. METHODS: 200 consecutive patients referred to a pain clinic for an epidural injection of steroid were randomly allocated to one of two groups. Group L had a lumbar approach to the epidural space and group C a caudal approach to the epidural space. Both groups then had epidurography performed using Omnipaque and an image intensifier to determine the position of the needle. RESULTS: Body mass index (BMI), grade of operator, and route of injection were predictors of a successful placement. 93% of lumbar and 64% of caudal epidural injections were correctly placed (p< 0.001). 97% of lumbar and 85% of caudal epidural injections clinically thought to be correctly placed were confirmed radiographically. For epidural injections where the clinical impression was "maybe", 91% of lumbar injections, but only 45% of caudal injections were correctly placed. Obesity was associated with a reduced chance of successful placement (odds ratio (OR) 0.34 (95% confidence interval (CI) 0.17 to 0.72) BMI >30 v BMI <30). A more senior grade of operator was associated with a reduced chance of successful placement (OR 0.16 (95% CI 0.03 to 0.89) consultant v other). However, small numbers may have accounted for the latter result. CONCLUSIONS: The weight of the patient and intended approach need to be considered when deciding the method used to enter the epidural space. In the non obese patient, lumbar epidural injections can be accurately placed without x ray screening, but caudal epidural injections, to be placed accurately, require x ray screening no matter what the weight of the patient. PMID- 11053066 TI - Recurrence risk modelling of the genetic susceptibility to ankylosing spondylitis. AB - OBJECTIVES: It has long been suspected that susceptibility to ankylosing spondylitis (AS) is influenced by genes lying distant to the major histocompatibility complex. This study compares genetic models of AS to assess the most likely mode of inheritance, using recurrence risk ratios in relatives of affected subjects. METHODS: Recurrence risk ratios in different degrees of relatives were determined using published data from studies specifically designed to address the question. The methods of Risch were used to determine the expected recurrence risk ratios in different degrees of relatives, assuming equal first degree relative recurrence risk between models. Goodness of fit was determined by chi(2) comparison of the expected number of affected subjects with the observed number, given equal numbers of each type of relative studied. RESULTS: The recurrence risks in different degrees of relatives were: monozygotic (MZ) twins 63% (17/27), first degree relatives 8.2% (441/5390), second degree relatives 1.0% (8/834), and third degree relatives 0. 7% (7/997). Parent-child recurrence risk (7.9%, 37/466) was not significantly different from the sibling recurrence risk (8.2%, 404/4924), excluding a significant dominance genetic component to susceptibility. Poor fitting models included single gene, genetic heterogeneity, additive, two locus multiplicative, and one locus and residual polygenes (chi(2) >32 (two degrees of freedom), p<10(-6) for all models). The best fitting model studied was a five locus model with multiplicative interaction between loci (chi(2)=1.4 (two degrees of freedom), p=0.5). Oligogenic multiplicative models were the best fitting over a range of population prevalences and first degree recurrence risk rates. CONCLUSIONS: This study suggests that of the genetic models tested, the most likely model operating in AS is an oligogenic model with predominantly multiplicative interaction between loci. PMID- 11053067 TI - Synovial fluid chondroitin sulphate epitopes 3B3 and 7D4, and glycosaminoglycan in human knee osteoarthritis after exercise. AB - OBJECTIVE: Walking exercise alleviates some symptoms, such as pain, in patients with mild to moderate knee osteoarthritis (OA). However, a major concern is that weightbearing exercise on osteoarthritic joints may exacerbate articular cartilage degradation. Loading of proteoglycan depleted articular cartilage in vitro increased expression of the chondroitin sulphate epitope 3B3, suggesting that loading may influence metabolism of osteoarthritic cartilage. This study aimed at evaluating the effects of walking exercise on articular cartilage metabolism in patients with knee OA, as reflected by changes in concentrations of synovial fluid markers. METHODS: Thirty elderly patients with knee OA (Kellgren Lawrence grades II to IV) were randomly allocated to control (n = 15) and 12 week exercise (n = 15) groups. Synovial fluid obtained from 21 of the patients at time zero and after 12 weeks was examined by enzyme linked immunosorbent assay (ELISA) for the chondroitin sulphate epitopes 3B3 and 7D4, and by a dye binding assay with 1, 9-dimethylmethylene blue for total sulphated glycosaminoglycan (GAG) concentrations. The 3B3/GAG and 7D4/GAG ratios were calculated. RESULTS: No significant changes in concentrations of 3B3, 7D4, GAG, 3B3/GAG, or 7D4/GAG between time zero and 12 weeks were found in either group. However, there were significant declines in 3B3 (p=0. 001), GAG (p=0.007), and the 3B3/GAG ratio (p=0.049) with aging. CONCLUSION: Twelve weeks of walking exercise had no demonstrable adverse effects on articular cartilage metabolism, as reflected by the concentrations of synovial fluid GAG or the chondroitin sulphate epitopes 3B3 and 7D4. PMID- 11053068 TI - Value of the time trade off method for measuring utilities in patients with rheumatoid arthritis. AB - OBJECTIVE: To assess the feasibility, reliability, and validity of the time trade off (TTO) in patients with rheumatoid arthritis (RA). METHODS: The TTO was applied in 194 patients with RA with increasing difficulty in performing activities of daily living. The test-retest reliability was determined in 35 of these patients and was calculated by the intraclass correlation coefficient (ICC). Construct validity was evaluated with the following sets of variables: measures of utility (rating scale), quality of life (RAND 36 item Health Status Survey (RAND-36) and RAQoL), functional status (Health Assessment Questionnaire, grip strength, and walk test), and disease activity (doctor's global assessment, disease activity score, pain, and morning stiffness). RESULTS: Ten patients (5%) did not complete the TTO. The median value of the TTO was 0.77 (range 0.03-1. 0). The test-retest ICC of the TTO was 0.85 (p<0.001). Construct validity testing of the TTO showed poor to moderate correlations (Spearman's r(s) between 0.19 and 0.36, p<0.01) with all outcome measures except for the subscale role limitation (physical problem) of the RAND-36, the walk test, the doctor's global assessment of disease activity, and morning stiffness. Multiple regression analysis showed that only 17% of the variance of the TTO scores could be explained. CONCLUSIONS: The TTO method appeared to be feasible and reliable in patients with RA. The poor to moderate correlations of the TTO with measures of quality of life, functional ability, and disease activity suggest that the TTO considers additional attributes of health status. This may have implications for the application of the TTO in clinical trials in patients with RA. PMID- 11053069 TI - Characterisation of autoantibodies to neutrophil granule constituents among patients with reactive arthritis, rheumatoid arthritis, and ulcerative colitis. AB - OBJECTIVE: To study the frequency and distribution of antineutrophil cytoplasmic autoantibodies (ANCA) among patients with reactive arthritis (ReA), rheumatoid arthritis (RA), and ulcerative colitis (UC) using different immunological methods. METHODS: Fifty serum samples from patients with reactive arthritis (26 with acute disease and 24 with chronic disease-that is disease of more than one year) were analysed for ANCA with indirect immunofluorescence, enzyme linked immunosorbent assay (ELISA) with six different neutrophil granule proteins as antigens, and immunoblotting on whole neutrophil extract and extracts of azurophil and specific granules. Thirty serum samples from patients with RA and UC served as controls in ELISA and indirect immunofluorescence. RESULTS: Sixteen per cent of patients with ReA were positive in immunofluorescence compared with 30% of patients with RA, and 70% of patients with UC. Thirty two per cent of patients with ReA were positive in ELISA. Antibodies directed against lactoferrin occurred in 20%, antibodies against bactericidal permeability increasing protein (BPI), elastase, cathepsin G, myeloperoxidase, and proteinase 3 were found in 8%, 2%, 2%, 8%, and 6%, respectively. Overall, 50% of RA sera and 53% of UC sera were positive in one or more ELISA assays, the corresponding figures for antibodies against individual antigens were for RA 7%, 3%, 0%, 13%, 47%, 17% and for UC 13%, 20%, 0%, 23%, 10%, and 17%. In immunoblotting, bands corresponding to lactoferrin and BPI were recognised in 44% and 22% of ReA sera. CONCLUSION: Antibodies against neutrophil granule antigens are often found in patients with ReA, primarily among those with chronic disease. The different methods detect various subsets of antibodies, with immunoblotting being the most and immunofluorescence the least sensitive. PMID- 11053070 TI - Lipoproteins and their subfractions in psoriatic arthritis: identification of an atherogenic profile with active joint disease. AB - OBJECTIVES: (a) To characterise the lipid profile in psoriatic arthritis and investigate whether there are similarities to the dyslipoproteinaemia reported in rheumatoid arthritis and other inflammatory forms of joint disease; (b) to investigate whether there is an atherogenic lipid profile in psoriatic arthritis, which may have a bearing on mortality. METHODS: Fasting lipids, lipoproteins, and their subfractions were measured in 50 patients with psoriatic arthritis and their age and sex matched controls. RESULTS: High density lipoprotein cholesterol (HDL cholesterol) and its third subfraction, HDL(3) cholesterol, were significantly reduced and the most dense subfraction of low density lipoprotein (LDL), LDL(3), was significantly increased in the patients with psoriatic arthritis. Twenty patients with active synovitis had significantly lower total cholesterol, LDL cholesterol, and HDL(3) cholesterol than their controls. 25% of the patients with psoriatic arthritis had raised Lp(a) lipoprotein levels (>300 mg/l) compared with 19% of controls, but this was not statistically significant. CONCLUSION: Raised levels of LDL(3) and low levels of HDL cholesterol are associated with coronary artery disease. Such an atherogenic profile in a chronic inflammatory form of arthritis is reported, which may be associated with accelerated mortality. PMID- 11053071 TI - A survey of phenotype II in familial Mediterranean fever. AB - OBJECTIVE: Phenotype II in familial Mediterranean fever (FMF) is the onset of amyloidosis before the onset of FMF with its typical attacks, or as an isolated finding in a member of an FMF family. Its presence was investigated by looking for proteinuria among the asymptomatic relatives of patients with FMF complicated by amyloidosis and among the asymptomatic relatives of patients with juvenile chronic arthritis (JCA) complicated by amyloidosis, used as controls. METHODS: The relatives of the index patients (13 with FMF and amyloidosis) and controls (6 with JCA and amyloidosis) were screened for proteinuria. Rectal biopsies were performed when proteinuria was significant (>/=300 mg/d). RESULTS: 461 relatives were screened in the FMF group and 269 among the controls. Two of the FMF relatives and one JCA relative had no symptoms of FMF but had significant proteinuria. Rectal biopsy for amyloidosis was negative in all instances of significant proteinuria. CONCLUSION: Phenotype II is uncommon among the relatives of patients with FMF and amyloidosis. PMID- 11053072 TI - Ciprofloxacin v placebo for treatment of Yersinia enterocolitica triggered reactive arthritis. AB - Patients with yersinia triggered reactive arthritis were double blind randomly allocated to receive treatment with ciprofloxacin 500 mg twice daily orally or placebo during three months. The diagnosis was made by serology (specific IgA and IgG antibodies to yersinia outer membrane proteins (yops)), positive culture, and/or demonstration of Yersinia enterocolitica antigen in colon biopsy specimens. Patients were evaluated monthly during and after treatment up to 12 months. Of 18 patients enrolled, all could be evaluated for safety, 16 for efficacy. There was a tendency towards faster remission and relief of pain in those receiving ciprofloxacin. Y enterocolitica was eliminated from the gut associated lymphoid tissue in six of seven patients receiving ciprofloxacin compared with none of nine patients receiving placebo. Patients receiving placebo had more and prolonged circulating IgA antibodies against yops than patients treated with ciprofloxacin. PMID- 11053073 TI - Corticosteroid injection for the treatment of carpal tunnel syndrome. AB - OBJECTIVE: To compare low and high dose, and short and long acting corticosteroids in the treatment of carpal tunnel syndrome. METHODS: A randomised, controlled, single blind trial with electromyographic and subjective outcome measures. RESULTS: 25 mg hydrocortisone is as effective as higher doses or long acting triamcinolone at a six week and six month follow up. CONCLUSION: As low dose steroid is as effective, and potentially less toxic, this should be the recommended dose for injection of carpal tunnel syndrome. PMID- 11053074 TI - Splenectomy for refractory Evans' syndrome associated with antiphospholipid antibodies: report of two cases. AB - The main haematological manifestations seen in patients with antiphospholipid antibodies (aPL) are thrombocytopenia, usually mild, and haemolytic anaemia with a positive Coombs test. Owing to the shared characteristics with idiopathic thrombocytopenic purpura, similar rules are followed in the treatment of these cytopenias. Two patients with severe aPL associated cytopenias, who required splenectomy after being refractory to steroids, immunosuppressive agents, and other treatments (intravenous gammaglobulin, danazol), are described, and previously reported cases are reviewed. PMID- 11053075 TI - What is new in systemic vasculitis? PMID- 11053077 TI - Updated consensus statement on tumour necrosis factor blocking agents for the treatment of rheumatoid arthritis (May 2000). PMID- 11053078 TI - Post-transcriptional regulation of tumour necrosis factor alpha production. PMID- 11053079 TI - Tumour necrosis factors receptor associated signalling molecules and their role in activation of apoptosis, JNK and NF-kappaB. AB - Tumour necrosis factor (TNF) is a pleiotropic cytokine that mediates apoptosis, cell proliferation, immunomodulation, inflammation, viral replication, allergy, arthritis, septic shock, insulin resistance, autoimmune diseases, and other pathological conditions. TNF transduces these cellular responses through two distinct receptors: type I, which are expressed on all cell types, and type II, which are expressed only on cells of the immune system and endothelial cells. At the cellular level, these receptors activate the pathways leading to the activation of transcription factors NF-kappaB and AP-1, apoptosis and proliferation, and mitogen activated protein kinases. None of these receptors exhibit any enzymatic activity but the signals are transmitted through the recruitment of more than a dozen different signalling proteins, which together form signalling cascades. Inhibitors of TNF signalling have therapeutic value as indicated by the approval of the soluble TNF receptors and anti-TNF antibodies for rheumatoid arthritis and for inflammatory bowl disease. PMID- 11053080 TI - Targeting interleukin 18 with interleukin 18 binding protein. AB - A novel, constitutively expressed and secreted interleukin 18 (IL18) binding protein (IL18BP) neutralises IL18. IL18BP shares many characteristics with soluble cytokine receptors of the IL1 family in that the protein exhibits specificity for IL18, belongs to the immunoglobulin-like class of receptors and has limited amino acid sequences with those of the IL1 receptor type II. However, unlike soluble cytokine receptors, IL18BP does not have a transmembrane domain and hence is not anchored to the cell membrane. IL18BP is a secreted protein and not cleaved from the cell surface. IL18BP is naturally occurring and was isolated from the urine of healthy subjects. Because IL18 is an important inducer of interferon gamma (IFNgamma), IL18BP suppresses the production of IFNgamma resulting in reduced T-helper type 1 immune responses. There are four human and two mouse isoforms-resulting from mRNA splicing and found in various cDNA libraries. Each of these IL18BP isoforms have been expressed, purified and assessed for binding and neutralisation of IL18 biological activities. Two human IL18BP isoforms exhibited the greatest affinity for IL18 with a rapid on-rate, a slow off-rate and a dissociation constant (kDa) of 399 pM. The two other isoforms with an incomplete immunoglobulin domain were unable to neutralise IL18. The two human isoforms that possess a complete immunoglobulin domain, neutralise >95% IL18 at a molar excess of two. Molecular modelling identified a large mixed electrostatic and hydrophobic binding site in the immunoglobulin domain of IL18BP, which could account for its high affinity binding to the ligand. These high affinity forms may be ideally suited for blocking IL18 in human disease. It is likely that preferential secretion of high affinity functional and non functional isoforms of IL18BP affect the immune response and the outcome of disease. PMID- 11053081 TI - Anti-interleukin 6 receptor antibody treatment in rheumatic disease. AB - Interleukin 6 (IL6) is a pleiotropic cytokine with a wide range of biological activities. IL6 transgene into mice gives rise to the abnormalities such as hypergammaglobulinaemia, thrombocytosis, infiltration of inflammatory cells into the tissues, mesangial cell proliferation of the kidney as well as splenomegaly and lymphadenopathy, which are predictable by the biological functions of IL6 shown in vitro. Continuous overproduction of IL6 is observed in patients with some immune-inflammatory diseases such as Castleman's disease and rheumatoid arthritis that are frequently associated with similar abnormalities to those of IL6 transgenic mice, strongly suggesting the involvement of IL6 in the human diseases. Successful treatment of the model animals for immune-inflammatory diseases with anti-IL6 receptor (IL6R) antibody thus indicates the possible application of IL6 blocking agents to treat the IL6 related immune-inflammatory diseases of humans. In this review, the new therapeutic strategy for Castleman's disease and RA using humanized antibody to human IL6 receptor, MRA, is discussed. PMID- 11053082 TI - Clinical outcome measures in rheumatoid arthritis. PMID- 11053083 TI - Efficacy and toxicity of old and new disease modifying antirheumatic drugs. PMID- 11053084 TI - Which post-marketing database and studies does EMEA require. PMID- 11053085 TI - The role of national agencies in the managed introduction of new drugs for arthritis. AB - The role of the various agencies involved in the managed introduction of new drugs in the United Kingdom is discussed, particularly with regard to the work of the National Institute for Clinical Excellence (NICE). The process by which the National Health Service in the UK identifies key new drug technologies of major clinical, financial and service significance is discussed. This includes the decision making process for selection of products for appraisal by NICE. The appraisal procedure and the impact of NICE guidance for the NHS will, it is hoped, encourage quality in clinical practice. All healthcare systems face budgetary constraints and the introduction of new technologies bring particular challenges. The launch of the new biological agents for the treatment of rheumatoid arthritis, into a speciality where previously relatively inexpensive agents were prescribed, raises questions concerning managed introduction and reimbursement. PMID- 11053086 TI - Treatment of rheumatoid arthritis with PEGylated recombinant human soluble tumour necrosis factor receptor type I: a clinical update. PMID- 11053087 TI - Update on D2E7: a fully human anti-tumour necrosis factor alpha monoclonal antibody. PMID- 11053088 TI - Etanercept (Enbrel): update on therapeutic use. AB - Tumour necrosis factor (TNF) is an important inflammatory disease mediator in a wide spectrum of articular diseases, including adult and juvenile rheumatoid arthritis (RA, JRA). Etanercept (Enbrel), approved in the United States and in Europe for use in patients with RA and JRA, is an effective inhibitor of TNF that has been shown to provide rapid and sustained improvement in both of these diseases. Long term studies continue to show that etanercept controls signs and symptoms of RA and JRA with no change in rate or type of adverse event over time. To demonstrate that etanercept is effective as first line treatment for patients with early active RA who have not been previously treated with methotrexate, and to examine the effect of etanercept on radiographic progression, a double blind, placebo controlled study was recently conducted, comparing etanercept with methotrexate (median dose 20 mg per week). Both etanercept 25 mg twice weekly and rapidly escalated methotrexate were effective in reducing the signs and symptoms of RA, and etanercept was significantly better than methotrexate in slowing the rate of radiographic erosions. In patients with severe psoriatic arthritis (PsA), a double blind, placebo controlled study demonstrated that etanercept was also effective in reducing disease activity in PsA. Etanercept has been well tolerated in all of these clinical trials and offers an important new treatment option to patients with inflammatory articular diseases. PMID- 11053089 TI - The pre-ligand binding assembly domain: a potential target of inhibition of tumour necrosis factor receptor function. AB - Signalling by the tumour necrosis factor receptors (TNFR) is thought to be mediated by the binding of the trimeric ligand TNF to three monomeric subunits of the receptor. This ligand induced trimerisation model of TNFR signalling is mainly supported by crystallographic data of the p60 TNFR-1 and TNFbeta complex in which the trimeric ligand interdigitates between the individual receptor chains and prevents the receptor subunits from interacting with each other. Recently, a domain NH(2)-terminal to the ligand binding domain in the extracellular region of p60 TNFR-1, p80 TNFR-2 and Fas was identified that mediates receptor self association before ligand binding. This pre-ligand binding assembly domain or PLAD is critical for assembly of functional receptor complexes on the cell surface and may provide a potential target in the design of future novel therapeutics against diseases mediated by members of the TNFR family of receptors. PMID- 11053090 TI - Adenoviral transgene delivery provides an approach to identifying important molecular processes in inflammation: evidence for heterogenecity in the requirement for NFkappaB in tumour necrosis factor production. AB - The success of anti-tumour necrosis factor (TNF) treatment, either using antibodies or soluble receptors, has defined TNF as a major factor of the inflammatory response in rheumatoid arthritis (RA). As a result of this success, much attention has been devoted to understanding the molecular mechanisms by which TNF expression and activity is elicited and controlled. By understanding these pathways, it is hoped that key elements of the molecular pathology associated with RA will be uncovered and, as a result, new targets for therapeutic intervention will be identified. However, studying the cell and molecular biology of model systems for RA, such as primary human macrophages, or tissue from rheumatoid joints may present technical problems. In an attempt to overcome this, we have investigated the use of adenovirus as a means of delivering transgenes by which different intracellular pathways can be modulated and examined. Our data show that adenovirus can be successfully used to efficiently deliver transgenes to primary human macrophages and RA joint tissue. Using a virus encoding IkappaBalpha, the natural inhibitor of NFkappaB, we show that the requirement for the transcription factor is not universal, but is dependent on the nature of the stimulus. Furthermore, while NFkappaB is of importance for the expression of TNF and other pro-inflammatory cytokines (for example, interleukin 6) and the destructive matrix metalloproteinases, this factor is not required for the expression of anti-inflammatory cytokines interleukin 10 and interleukin 1 receptor antagonist. PMID- 11053091 TI - Biological role of interleukin 1 receptor antagonist isoforms. AB - The interleukin 1 receptor antagonist (IL1Ra) family of molecules now includes one secreted isoform (sIL1Ra) and three intracellular isoforms (icIL1Ra1, 2, and 3). Extensive evidence indicates that the sole biological function of sIL1Ra seems to be to competitively inhibit IL1 binding to cell-surface receptors. Although intracellular IL1Ra1 may be released from keratinocytes under some conditions, the intracellular isoforms of IL1Ra may carry out additional as yet poorly defined roles inside cells. Maintenance of a balance between IL1 and IL1Ra is important in preventing the development or progression of inflammatory disease in certain organs. Both the secreted and intracellular isoforms of IL1Ra contribute to maintenance of this balance. An allelic polymorphism in intron 2 of the IL1Ra gene (IL1RN*2) predisposes to the development or severity of a variety of human diseases largely of epithelial cell origin. Both the impaired production of IL1Ra and the overproduction of IL1beta are related to the presence of this allele. Restoration of the balance between IL1Ra and IL1 through a variety of approaches is a therapeutic goal in specific chronic inflammatory diseases. PMID- 11053092 TI - The role of angiogenesis in rheumatoid arthritis: recent developments. PMID- 11053093 TI - The final pathogenetic steps in focal bone erosions in rheumatoid arthritis. PMID- 11053094 TI - Cartilage destruction and bone erosion in arthritis: the role of tumour necrosis factor alpha. PMID- 11053095 TI - Arguments for interleukin 1 as a target in chronic arthritis. AB - Tumour necrosis factor (TNF) and interleukin 1 (IL1) are considered as master cytokines in chronic, destructive arthritis. Therapeutic approaches in rheumatic arthritis (RA) patients so far mainly focused on TNF. Although TNF is a major inflammatory mediator in RA and a potent inducer of IL1, anti-TNF treatment is not effective in all patients, nor does it fully control the arthritic process in affected joints of good responders. Analysis of cytokine patterns in early synovial biopsy specimens of RA patients reveals prominent TNF staining in 50% of the patients, whereas IL1b staining was evident in 100%. This argues that TNF independent IL1 production occurs in some of the patients. Studies in a range of experimental arthritis models in mice make it clear that TNF is involved in early joint swelling. However, TNF alone is not arthritogenic nor destructive and exerts its arthritogenic potential through IL1 induction. Intriguingly, TNF independent IL1 production is found in many models. Its relevance is further underlined by the greater efficacy of anti-IL1 treatment as compared with anti TNF treatment and the total lack of chronic, erosive arthritis in IL1b deficient mice. IL1b is not necessarily involved in early joint swelling, but is a crucial mediator in chronic arthritis and cartilage erosion in all models studied so far. This makes ILb an attractive target in chronic, destructive arthritis. PMID- 11053096 TI - Treatment of spondyloarthropathies with antibodies against tumour necrosis factor alpha: first clinical and laboratory experiences. PMID- 11053097 TI - Activation of cytokines as a mechanism of disease progression in heart failure. PMID- 11053098 TI - Tumour necrosis factor and anti-tumour necrosis factor approach to inflammatory demyelinating diseases of the central nervous system. PMID- 11053099 TI - Role of interleukin 1 and interleukin 1 receptor antagonist in the mediation of rheumatoid arthritis. AB - Chronic arthritis is characterised by chronic joint inflammation and concurrent joint erosion and destruction. The inflammatory cytokine interleukin 1 (IL1) has been shown to be a key mediator in the autoimmune disease rheumatoid arthritis (RA). Interleukin 1 mediates bone resorption and cartilage destruction, but may not play as dominant a part in joint swelling and inflammation. Interleukin 1 receptor antagonist (IL1Ra) selectively inhibits the effects of IL1 by competing for the IL1 receptor on all surfaces of the synovium. In a randomised controlled trial in 472 patients with active disease, IL1Ra 30 mg/day, 75 mg/day or 150 mg/day given by subcutaneous injection significantly reduced the signs and symptoms of RA at 24 weeks. An American College of Rheumatology (ACR) 20% response was seen in 43% of the patients treated with 150 mg/day at 24 weeks. IL1Ra was well tolerated; injection site reactions were the most common adverse event. In another trial, in 419 patients with active RA treated concomitantly with methotrexate, there were ACR 20% responses after 24 weeks in 42% of the patients treated with 1 mg/kg/day by subcutaneous injection and in 35% of those treated with 2 mg/kg/day. I1Ra offers a unique selective, targeted mechanism of action to block the IL1 mediated effects of RA. PMID- 11053100 TI - Advances in interleukin 2 receptor targeted treatment. AB - T cell activation and cellular immune responses are modulated by interleukin 2 (IL2) through binding to its corresponding cell surface receptor. Three forms of the receptor are recognised based on IL2 binding affinity. The high affinity receptor is a heterotrimer composed of alpha, beta, and gamma(c)-polypeptide chains. The 55 kDa alpha-chain also known as the Tac (T cell activation) antigen or CD-25 is a unique subunit of the high affinity IL2 receptor (IL2Ralpha). Resting T cells express few IL2Ralpha, however, when activated, the expression of ILR2alpha rapidly increases. The IL2Ralpha is shed from the cell surface and is measurable in the serum as a 45 kDa soluble form (s-Tac or s-IL2Ralpha). Serum concentrations of s-Tac can be used as a surrogate marker for T cell activation and IL2Ralpha expression. IL2Ralpha is over expressed by T cells in a number of autoimmune diseases, allograft rejection and a variety of lymphoid neoplasms. IL2 induced proliferation of T cells can be inhibited by the murine monoclonal antibody (anti-Tac) directed against the alpha-chain of the IL2R. Through molecular engineering, murine anti-Tac has been humanised reducing its immunogenicity without changing its specificity. Humanised anti-Tac (HAT) has been shown to reduce the incidence of renal and cardiac allograft rejection as well as decrease the severity of graft versus host disease in patients undergoing HLA matched allogeneic bone marrow transplantation. IL2Ralpha targeted treatment with radioimmunoconjugates of anti-Tac and immunotoxins has shown promise in the treatment of CD25 expressing lymphomas. PMID- 11053101 TI - Jaks and stats as therapeutic targets. AB - Cytokines have critical functions in regulating immune responses. A large number of these factors bind related receptors termed the Type I and Type II families of cytokine receptors. These receptors activate Janus kinases (Jaks) and Stat family of transcription factors. The essential and specific function of Jaks and Stats is particularly well illustrated by human and mouse mutations. The possibility that these molecules could be targeted to produce novel immunosuppressive compounds is considered in this review. PMID- 11053102 TI - Future prospects for anti-cytokine treatment. AB - The era of anti-cytokine treatment in rheumatology has just begun. The first generation therapeutic agents, biological agents that block tumour necrosis factor alpha such as monoclonal antibodies or receptor Ig fusion proteins are safe and effective, and so this has generated much interest in how to increase the benefit or deliver it more cost effectively. This article provides a personal view of the coming trends in anti-cytokine treatment. Which of these will be realised in the future will be of interest. PMID- 11053103 TI - Bistability in the Ca(2+)/calmodulin-dependent protein kinase-phosphatase system. AB - A mathematical model is presented of autophosphorylation of Ca(2+)/calmodulin dependent protein kinase (CaMKII) and its dephosphorylation by a phosphatase. If the total concentration of CaMKII subunits is significantly higher than the phosphatase Michaelis constant, two stable steady states of the CaMKII autophosphorylation can exist in a Ca(2+) concentration range from below the resting value of the intracellular [Ca(2+)] to the threshold concentration for induction of long-term potentiation (LTP). Bistability is a robust phenomenon, it occurs over a wide range of parameters of the model. Ca(2+) transients that switch CaMKII from the low-phosphorylated state to the high-phosphorylated one are in the same range of amplitudes and frequencies as the Ca(2+) transients that induce LTP. These results show that the CaMKII-phosphatase bistability may play an important role in long-term synaptic modifications. They also suggest a plausible explanation for the very high concentrations of CaMKII found in postsynaptic densities of cerebral neurons. PMID- 11053104 TI - Excluded volume in solvation: sensitivity of scaled-particle theory to solvent size and density. AB - Changes in solvent environment greatly affect macromolecular structure and stability. To investigate the role of excluded volume in solvation, scaled particle theory is often used to calculate delta G(tr)(ev), the excluded-volume portion of the solute transfer free energy, delta G(tr). The inputs to SPT are the solvent radii and molarities. Real molecules are not spheres. Hence, molecular radii are not uniquely defined and vary for any given species. Since delta G(tr)(ev) is extremely sensitive to solvent radii, uncertainty in these radii causes a large uncertainty in delta G(tr)(ev)-several kcal/mol for amino acid solutes transferring from water to aqueous mixtures. This uncertainty is larger than the experimental delta G(tr) values. Also, delta G(tr)(ev) can be either positive or negative. Adding neutral crowding molecules may not necessarily reduce solubility. Lastly, delta G(tr)(ev) is very sensitive to solvent density, rho. A few percent error in rho may even cause qualitative deviations in delta G(tr)(ev). For example, if rho is calculated by assuming the hard-sphere pressure to be constant, then delta G(tr)(ev) values and uncertainties are now only tenths of a kcal/mol and are positive. Because delta G(tr)(ev) values calculated by scaled-particle theory are strongly sensitive to solvent radii and densities, determining the excluded-volume contribution to transfer free energies using SPT may be problematic. PMID- 11053105 TI - Models of post-translational protein translocation. AB - Organellar Hsp-70 is required for post-translational translocation into the endoplasmic reticulum and mitochondria. The functional role played by Hsp-70 is unknown. However, two operating principles have been suggested. The power stroke model proposes that Hsp-70 undergoes a conformational change, which pulls the precursor protein through the translocation pore, whereas, in the Brownian ratchet model, the role of Hsp-70 is simply to block backsliding through the pore. A mathematical analysis of both mechanisms is presented and reveals that qualitative differences between the models occur in the behavior of the mean velocity and effective diffusion coefficient as a function of Hsp-70 concentration. An experimental method is proposed for measuring these two quantities that only relies on current experimental techniques. PMID- 11053106 TI - On hydrophobicity correlations in protein chains. AB - We study the statistical properties of hydrophobic/polar model sequences with unique native states on the square lattice. It is shown that this ensemble of sequences differs from random sequences in significant ways in terms of both the distribution of hydrophobicity along the chains and total hydrophobicity. Whenever statistically feasible, the analogous calculations are performed for a set of real enzymes, too. PMID- 11053107 TI - An elastic analysis of Listeria monocytogenes propulsion. AB - The bacterium Listeria monocytogenes uses the energy of the actin polymerization to propel itself through infected tissues. In steady state, it continuously adds new polymerized filaments to its surface, pushing on its tail, which is made from previously cross-linked actin filaments. In this paper we introduce an elastic model to describe how the addition of actin filaments to the tail results in the propulsive force on the bacterium. Filament growth on the bacterial surface produces stresses that are relieved at the back of the bacterium as it moves forward. The model leads to a natural competition between growth from the sides and growth from the back of the bacterium, with different velocities and strengths for each. This competition can lead to the periodic motion observed in a Listeria mutant. PMID- 11053108 TI - Molecular dynamics of the anticodon domain of yeast tRNA(Phe): codon-anticodon interaction. AB - We have studied the effect of codon-anticodon interaction on the structure and dynamics of transfer RNAs using molecular dynamics simulations over a nanosecond time scale. From our molecular dynamical investigations of the solvated anticodon domain of yeast tRNA(Phe) in the presence and absence of the codon trinucleotides UUC and UUU, we find that, although at a gross level the structures are quite similar for the free and the bound domains, there are small but distinct differences in certain parts of the molecule, notably near the Y37 base. Comparison of the dynamics in terms of interatomic or inter-residual distance fluctuation for the free and the bound domains showed regions of enhanced rigidity in the loop region in the presence of codons. Because fluorescence experiments suggested the existence of multiple conformers of the anticodon domain, which interconvert on a much larger time scale than our simulations, we probed the conformational space using five independent trajectories of 500 ps duration. A generalized ergodic measure analysis of the trajectories revealed that at least for this time scale, all the trajectories populated separate parts of the conformational space, indicating a need for even longer simulations or enhanced sampling of the conformational space to give an unequivocal answer to this question. PMID- 11053109 TI - Effect of overall feedback inhibition in unbranched biosynthetic pathways. AB - We have determined the effects of control by overall feedback inhibition on the systemic behavior of unbranched metabolic pathways with an arbitrary pattern of other feedback inhibitions by using a recently developed numerical generalization of Mathematically Controlled Comparisons, a method for comparing the function of alternative molecular designs. This method allows the rigorous determination of the changes in systemic properties that can be exclusively attributed to overall feedback inhibition. Analytical results show that the unbranched pathway can achieve the same steady-state flux, concentrations, and logarithmic gains with respect to changes in substrate, with or without overall feedback inhibition. The analytical approach also shows that control by overall feedback inhibition amplifies the regulation of flux by the demand for end product while attenuating the sensitivity of the concentrations to the same demand. This approach does not provide a clear answer regarding the effect of overall feedback inhibition on the robustness, stability, and transient time of the pathway. However, the generalized numerical method we have used does clarify the answers to these questions. On average, an unbranched pathway with control by overall feedback inhibition is less sensitive to perturbations in the values of the parameters that define the system. The difference in robustness can range from a few percent to fifty percent or more, depending on the length of the pathway and on the metabolite one considers. On average, overall feedback inhibition decreases the stability margins by a minimal amount (typically less than 5%). Finally, and again on average, stable systems with overall feedback inhibition respond faster to fluctuations in the metabolite concentrations. Taken together, these results show that control by overall feedback inhibition confers several functional advantages upon unbranched pathways. These advantages provide a rationale for the prevalence of this control mechanism in unbranched metabolic pathways in vivo. PMID- 11053110 TI - Function-related regulation of the stability of MHC proteins. AB - Proteins must be stable to accomplish their biological function and to avoid enzymatic degradation. Constitutive proteolysis, however, is the main source of free amino acids used for de novo protein synthesis. In this paper the delicate balance of protein stability and degradability is discussed in the context of function of major histocompatibility complex (MHC) encoded protein. Classical MHC proteins are single-use peptide transporters that carry proteolytic degradation products to the cell surface for presenting them to T cells. These proteins fulfill their function as long as they bind their dissociable ligand, the peptide. Ligand-free MHC molecules on the cell surface are practically useless for their primary biological function, but may acquire novel activity or become an important source of amino acids when they lose their compact stable structure, which resists proteolytic attacks. We show in this paper that one or more of the stabilization centers responsible for the stability of MHC-peptide complexes is composed of residues of both the protein and the peptide, therefore missing in the ligand-free protein. This arrangement of stabilization centers provides a simple means of regulation; it makes the useful form of the protein stable, whereas the useless form of the same protein is unstable and therefore degradable. PMID- 11053111 TI - Depletion theory of protein transport in semi-dilute polymer solutions. AB - We consider the effect of polymer depletion on the transport (diffusion and electrophoresis) of small proteins through semi-dilute solutions of a flexible polymer. A self-consistent field theory may be set up in the important case of quasi-ideal interactions when the protein is small enough. Dynamic depletion, the reorganization of the depletion layer as the protein diffuses, is computed within a free-draining approximation. The transport of the dressed particle (protein + depletion layer) is tackled by extending Ogston's analysis of probe diffusion through fibrous networks to the case of a probe diffusing through a semi-dilute polymer inhomogeneous on the scale of the polymer correlation length. The resulting exponential retardation agrees almost quantitatively with that found in recent electrophoresis experiments of small proteins in polymer solutions that have been ascertained to be semi-dilute (S. P. Radko and A. Chrambach, Electrophoresis, 17:1094-1102, 1996; Biopolymers, 4:183-189, 1997). PMID- 11053112 TI - Calculations suggest a pathway for the transverse diffusion of a hydrophobic peptide across a lipid bilayer. AB - Alamethicin is a hydrophobic antibiotic peptide 20 amino acids in length. It is predominantly helical and partitions into lipid bilayers mostly in transmembrane orientations. The rate of the peptide transverse diffusion (flip-flop) in palmitoyl-oleyl-phosphatidylcholine vesicles has been measured recently and the results suggest that it involves an energy barrier, presumably due to the free energy of transfer of the peptide termini across the bilayer. We used continuum solvent model calculations, the known x-ray crystal structure of alamethicin and a simplified representation of the lipid bilayer as a slab of low dielectric constant to calculate the flip-flop rate. We assumed that the lipids adjust rapidly to each configuration of alamethicin in the bilayer because their motions are significantly faster than the average peptide flip-flop time. Thus, we considered the process as a sequence of discrete peptide-membrane configurations, representing critical steps in the diffusion, and estimated the transmembrane flip-flop rate from the calculated free energy of the system in each configuration. Our calculations indicate that the simplest possible pathway, i.e., the rotation of the helix around the bilayer midplane, involving the simultaneous burial of the two termini in the membrane, is energetically unfavorable. The most plausible alternative is a two-step process, comprised of a rotation of alamethicin around its C-terminus residue from the initial transmembrane orientation to a surface orientation, followed by a rotation around the N-terminus residue from the surface to the final reversed transmembrane orientation. This process involves the burial of one terminus at a time and is much more likely than the rotation of the helix around the bilayer midplane. Our calculations give flip-flop rates of approximately 10(-7)/s for this pathway, in accord with the measured value of 1.7 x 10(-6)/s. PMID- 11053113 TI - Molecular dynamics simulations predict a tilted orientation for the helical region of dynorphin A(1-17) in dimyristoylphosphatidylcholine bilayers. AB - The structural properties of the endogenous opioid peptide dynorphin A(1-17) (DynA), a potential analgesic, were studied with molecular dynamics simulations in dimyristoylphosphatidylcholine bilayers. Starting with the known NMR structure of the peptide in dodecylphosphocholine micelles, the N-terminal helical segment of DynA (encompassing residues 1-10) was initially inserted in the bilayer in a perpendicular orientation with respect to the membrane plane. Parallel simulations were carried out from two starting structures, systems A and B, that differ by 4 A in the vertical positioning of the peptide helix. The complex consisted of approximately 26,400 atoms (dynorphin + 86 lipids + approximately 5300 waters). After >2 ns of simulation, which included >1 ns of equilibration, the orientation of the helical segment of DynA had undergone a transition from parallel to tilted with respect to the bilayer normal in both the A and B systems. When the helix axis achieved a approximately 50 degrees angle with the bilayer normal, it remained stable for the next 1 ns of simulation. The two simulations with different starting points converged to the same final structure, with the helix inserted in the bilayer throughout the simulations. Analysis shows that the tilted orientation adopted by the N-terminal helix is due to specific interactions of residues in the DynA sequence with phospholipid headgroups, water, and the hydrocarbon chains. Key elements are the "snorkel model"-type interactions of arginine side chains, the stabilization of the N-terminal hydrophobic sequence in the lipid environment, and the specific interactions of the first residue, Tyr. Water penetration within the bilayer is facilitated by the immersed DynA, but it is not uniform around the surface of the helix. Many water molecules surround the arginine side chains, while water penetration near the helical surface formed by hydrophobic residues is negligible. A mechanism of receptor interaction is proposed for DynA, involving the tilted orientation observed from these simulations of the peptide in the lipid bilayer. PMID- 11053114 TI - GFP is a selective non-linear optical sensor of electrophysiological processes in Caenorhabditis elegans. AB - Electrophysiology of the nematode Caenorhabditis elegans has the potential to bridge the wealth of information on the molecular biology and anatomy of this organism with the responses of selected cells and cellular neural networks associated with a behavioral response. In this paper we report that the nonlinear optical phenomenon of second harmonic generation (SHG) can be detected using green fluorescent protein (GFP) chimeras expressed in selected cells of living animals. Alterations in the SHG signal as a result of receptor ligand interactions and mechanical stimulation of the mechanosensory cells indicate that this signal is very sensitive to membrane potential. The results suggest that this approach to membrane potential measurements in C. elegans and in other biological systems could effectively couple data on selective locations within specific cells with functional responses that are associated with behavioral and sensory processes. PMID- 11053115 TI - Cell mechanics studied by a reconstituted model tissue. AB - Tissue models reconstituted from cells and extracellular matrix (ECM) simulate natural tissues. Cytoskeletal and matrix proteins govern the force exerted by a tissue and its stiffness. Cells regulate cytoskeletal structure and remodel ECM to produce mechanical changes during tissue development and wound healing. Characterization and control of mechanical properties of reconstituted tissues are essential for tissue engineering applications. We have quantitatively characterized mechanical properties of connective tissue models, fibroblast populated matrices (FPMs), via uniaxial stretch measurements. FPMs resemble natural tissues in their exponential dependence of stress on strain and linear dependence of stiffness on force at a given strain. Activating cellular contractile forces by calf serum and disrupting F-actin by cytochalasin D yield "active" and "passive" components, which respectively emphasize cellular and matrix mechanical contributions. The strain-dependent stress and elastic modulus of the active component were independent of cell density above a threshold density. The same quantities for the passive component increased with cell number due to compression and reorganization of the matrix by the cells. PMID- 11053116 TI - A cell-based constitutive relation for bio-artificial tissues. AB - By using a combination of continuum and statistical mechanics we derive an integral constitutive relation for bio-artificial tissue models consisting of a monodisperse population of cells in a uniform collagenous matrix. This constitutive relation quantitatively models the dependence of tissue stress on deformation history, and makes explicit the separate contribution of cells and matrix to the mechanical behavior of the composite tissue. Thus microscopic cell mechanical properties can be deduced via this theory from measurements of macroscopic tissue properties. A central feature of the constitutive relation is the appearance of "anisotropy tensors" that embody the effects of cell orientation on tissue mechanics. The theory assumes that the tissues are stable over the observation time, and does not in its present form allow for cell migration, reorientation, or internal remodeling. We have compared the predictions of the theory to uniaxial relaxation tests on fibroblast-populated collagen matrices (FPMs) and find that the experimental results generally support the theory and yield values of fibroblast contractile force and stiffness roughly an order of magnitude smaller than, and viscosity comparable to, the corresponding properties of active skeletal muscle. The method used here to derive the tissue constitutive equation permits more sophisticated cell models to be used in developing more accurate representations of tissue properties. PMID- 11053117 TI - Mapping the growth of fungal hyphae: orthogonal cell wall expansion during tip growth and the role of turgor. AB - By computer-enhanced videomicroscopy, we mapped the trajectory of external and internal cell surface markers in growing fungal hyphae to determine the pattern of cell wall expansion during apical growth. Carbon particles (India ink) were chosen as external markers for tip expansion of Rhizoctonia solani hyphae. Irregularities in the growing apical walls of R. solani served as internal markers. Marker movement was traced in captured frames from the videotaped sequences. External and internal markers both followed orthogonal trajectories; i.e., they moved perpendicular to the cell surface regardless of their initial position in the hyphal apex. We found no evidence that the tip rotates during elongation. The discovery that the cell wall of a growing hypha expands orthogonally has major repercussions on two fronts: 1) It supports the long-held view that turgor pressure is the main force driving cell wall expansion. 2) It provides crucial information to complete the mathematical derivation of a three dimensional model of hyphal morphogenesis based on the vesicle supply center concept. In three dimensions, the vesicle gradient generated by the vesicle supply center is insufficient to explain shape; it is also necessary to know the manner in which the existing surface is displaced during wall expansion. PMID- 11053118 TI - Cell-free rolling mediated by L-selectin and sialyl Lewis(x) reveals the shear threshold effect. AB - The selectin family of adhesion molecules mediates attachment and rolling of neutrophils to stimulated endothelial cells. This step of the inflammatory response is a prerequisite to firm attachment and extravasation. We have reported that microspheres coated with sialyl Lewis(x) (sLe(x)) interact specifically and roll over E-selectin and P-selectin substrates (Brunk et al., 1996; Rodgers et al 2000). This paper extends the use of the cell-free system to the study of the interactions between L-selectin and sLe(x) under flow. We find that sLe(x) microspheres specifically interact with and roll on L-selectin substrates. Rolling velocity increases with wall shear stress and decreases with increasing L selectin density. Rolling velocities are fast, between 25 and 225 microm/s, typical of L-selectin interactions. The variability of rolling velocity, quantified by the variance in rolling velocity, scales linearly with rolling velocity. Rolling flux varies with both wall shear stress and L-selectin site density. At a density of L-selectin of 800 sites/microm(2), the rolling flux of sLe(x) coated microspheres goes through a clear maximum with respect to shear stress at 0.7 dyne/cm(2). This behavior, in which the maintenance and promotion of rolling interactions on selectins requires shear stress above a threshold value, is known as the shear threshold effect. We found that the magnitude of the effect is greatest at an L-selectin density of 800 sites/microm(2) and gradually diminishes with increasing L-selectin site density. Our study is the first to reveal the shear threshold effect with a cell free system and the first to show the dependence of the shear threshold effect on L-selectin site density in a reconstituted system. Our ability to recreate the shear threshold effect in a cell-free system strongly suggests the origin of the effect is in the physical chemistry of L-selectin interaction with its ligand, and largely eliminates cellular features such as deformability or topography as its cause. PMID- 11053119 TI - Reversal of the gating polarity of gap junctions by negative charge substitutions in the N-terminus of connexin 32. AB - Intercellular channels formed by connexins (gap junctions) are sensitive to the application of transjunctional voltage (V(j)), to which they gate by the separate actions of their serially arranged hemichannels (Harris, A. L., D. C. Spray, and M. V. L. Bennett. 1981. J. Gen. Physiol. 77:95-117). Single channel studies of both intercellular and conductive hemichannels have demonstrated the existence of two separate gating mechanisms, termed "V(j)-gating" and "loop gating" (Trexler, E. B., M. V. L. Bennett, T. A. Bargiello, and V. K. Verselis. 1996. Proc. Natl. Acad. Sci. U.S.A. 93:5836-5841). In Cx32 hemichannels, V(j)-gating occurs at negative V(j) (Oh, S., J. B. Rubin, M. V. L. Bennett, V. K. Verselis, and T. A. Bargiello. 1999. J. Gen. Physiol. 114:339-364; Oh, S., C. K. Abrams, V. K. Verselis, and T. A. Bargiello. 2000. J. Gen. Physiol. 116:13-31). A negative charge substitution at the second amino acid position in the N-terminus reverses the polarity of V(j)-gating of Cx32 hemichannels (Verselis, V. K., C. S. Ginter, and T. A. Bargiello. 1994. Nature. 368:348-351;. J. Gen. Physiol. 116:13-31). We report that placement of a negative charge at the 5th, 8th, 9th, or 10th position can reverse the polarity of Cx32 hemichannel V(j)-gating. We conclude that the 1st through 10th amino acid residues lie within the transjunctional electric field and within the channel pore, as in this position they could sense changes in V(j) and be largely insensitive to changes in absolute membrane potential (V(m)). Conductive hemichannels formed by Cx32*Cx43E1 containing a negatively charged residue at either the 8th or 10th position display bi-polar V(j)-gating; that is, the open probability of hemichannels formed by these connexins is reduced at both positive and negative potentials and is maximal at intermediate voltages. In contrast, Cx32*Cx43E1 hemichannels with negative charges at either the 2nd or 5th positions are uni-polar, closing only at positive V(j). The simplest interpretation of these data is that the Cx32 hemichannel can adopt at least two different open conformations. The 1st-5th residues are located within the electric field in all open channel conformations, while the 8th and 10th residues lie within the electric field in one conformation and outside the electric field in the other conformation. PMID- 11053120 TI - A high-Na(+) conduction state during recovery from inactivation in the K(+) channel Kv1.5. AB - Na(+) conductance through cloned K(+) channels has previously allowed characterization of inactivation and K(+) binding within the pore, and here we have used Na(+) permeation to study recovery from C-type inactivation in human Kv1.5 channels. Replacing K(+) in the solutions with Na(+) allows complete Kv1.5 inactivation and alters the recovery. The inactivated state is nonconducting for K(+) but has a Na(+) conductance of 13% of the open state. During recovery, inactivated channels progress to a higher Na(+) conductance state (R) in a voltage-dependent manner before deactivating to closed-inactivated states. Channels finally recover from inactivation in the closed configuration. In the R state channels can be reactivated and exhibit supernormal Na(+) currents with a slow biexponential inactivation. Results suggest two pathways for entry to the inactivated state and a pore conformation, perhaps with a higher Na(+) affinity than the open state. The rate of recovery from inactivation is modulated by Na(+)(o) such that 135 mM Na(+)(o) promotes the recovery to normal closed, rather than closed-inactivated states. A kinetic model of recovery that assumes a highly Na(+)-permeable state and deactivation to closed-inactivated and normal closed states at negative voltages can account for the results. Thus these data offer insight into how Kv1. 5 channels recover their resting conformation after inactivation and how ionic conditions can modify recovery rates and pathways. PMID- 11053121 TI - Variable ratio of permeability to gating charge of rBIIA sodium channels and sodium influx in Xenopus oocytes. AB - Whole-cell gating current recording from rat brain IIA sodium channels in Xenopus oocytes was achieved using a high-expression system and a newly developed high speed two-electrode voltage-clamp. The resulting ionic currents were increased by an order of magnitude. Surprisingly, the measured corresponding gating currents were approximately 5-10 times larger than expected from ionic permeability. This prompted us to minimize uncertainties about clamp asymmetries and to quantify the ratio of sodium permeability to gating charge, which initially would be expected to be constant for a homogeneous channel population. The systematic study, however, showed a 10- to 20-fold variation of this ratio in different experiments, and even in the same cell during an experiment. The ratio of P(Na)/Q was found to correlate with substantial changes observed for the sodium reversal potential. The data suggest that a cytoplasmic sodium load in Xenopus oocytes or the energy consumption required to regulate the increase in cytoplasmic sodium represents a condition where most of the expressed sodium channels keep their pore closed due to yet unknown mechanisms. In contrast, the movements of the voltage sensors remain undisturbed, producing gating current with normal properties. PMID- 11053122 TI - NMDA channel gating is influenced by a tryptophan residue in the M2 domain but calcium permeation is not altered. AB - N-Methyl-D-aspartate (NMDA) receptors are susceptible to open-channel block by dizolcipine (MK-801), ketamine and Mg(2+) and are permeable to Ca(2+). It is thought that a tryptophan residue in the second membrane-associated domain (M2) may form part of the binding site for open-channel blockers and contribute to Ca(2+) permeability. We tested this hypothesis using recombinant wild-type and mutant NMDA receptors expressed in HEK-293 cells. The tryptophan was mutated to a leucine (W-5L) in both the NMDAR1 and NMDAR2A subunits. MK-801 and ketamine progressively inhibited currents evoked by glutamate, and the rate of inhibition was increased by the W-5L mutation. An increase in open channel probability accounted for the acceleration. Fluctuation analysis of the glutamate-evoked current revealed that the NMDAR1 W-5L mutation increased channel mean open time, providing further evidence for an alteration in gating. However, the equilibrium affinities of Mg(2+) and ketamine were largely unaffected by the W-5L mutation, and Ca(2+) permeability was not decreased. Therefore, the M2 tryptophan residue of the NMDA channel is not involved in Ca(2+) permeation or the binding of open channel blockers, but plays an important role in channel gating. PMID- 11053123 TI - Plasmon resonance studies of agonist/antagonist binding to the human delta-opioid receptor: new structural insights into receptor-ligand interactions. AB - Structural changes accompanying the binding of ligands to the cloned human delta opioid receptor immobilized in a solid-supported lipid bilayer have been investigated using coupled plasmon-waveguide resonance spectroscopy. This highly sensitive technique directly monitors mass density, conformation, and molecular orientation changes occurring in anisotropic thin films and allows direct determination of binding constants. Although both agonist binding and antagonist binding to the receptor cause increases in molecular ordering within the proteolipid membrane, only agonist binding induces an increase in thickness and molecular packing density of the membrane. This is a consequence of mass movements perpendicular to the plane of the bilayer occurring within the lipid and receptor components. These results are consistent with models of receptor function that involve changes in the orientation of transmembrane helices. PMID- 11053124 TI - The interaction of Na(+) and K(+) in the pore of cyclic nucleotide-gated channels. AB - The permeability ratio between K(+) and Na(+) ions in cyclic nucleotide-gated channels is close to 1, and the single channel conductance has almost the same value in the presence of K(+) or Na(+). Therefore, K(+) and Na(+) ions are thought to permeate with identical properties. In the alpha-subunit from bovine rods there is a loop of three prolines at positions 365 to 367. When proline 365 is mutated to a threonine, a cysteine, or an alanine, mutant channels exhibit a complex interaction between K(+) and Na(+) ions. Indeed K(+), Rb(+) and Cs(+) ions do not carry any significant macroscopic current through mutant channels P365T, P365C and P365A and block the current carried by Na(+) ions. Moreover in mutant P365T the presence of K(+) in the intracellular (or extracellular) medium caused the appearance of a large transient inward (or outward) current carried by Na(+) when the voltage command was quickly stepped to large negative (or positive) membrane voltages. This transient current is caused by a transient potentiation, i.e., an increase of the open probability. The permeation of organic cations through these mutant channels is almost identical to that through the wild type (w.t.) channel. Also in the w.t. channel a similar but smaller transient current is observed, associated to a slowing down of the channel gating evident when intracellular Na(+) is replaced with K(+). As a consequence, a rather simple mechanism can explain the complex behavior here described: when a K(+) ion is occupying the pore there is a profound blockage of the channel and a potentiation of gating immediately after the K(+) ion is driven out. Potentiation occurs because K(+) ions slow down the rate constant K(off) controlling channel closure. These results indicate that K(+) and Na(+) ions do not permeate through CNG channels in the same way and that K(+) ions influence the channel gating. PMID- 11053125 TI - RYR1 and RYR3 have different roles in the assembly of calcium release units of skeletal muscle. AB - Calcium release units (CRUs) are junctions between the sarcoplasmic reticulum (SR) and exterior membranes that mediates excitation contraction (e-c) coupling in muscle cells. In skeletal muscle CRUs contain two isoforms of the sarcoplasmic reticulum Ca(2+)release channel: ryanodine receptors type 1 and type 3 (RyR1 and RyR3). 1B5s are a mouse skeletal muscle cell line that carries a null mutation for RyR1 and does not express either RyR1 or RyR3. These cells develop dyspedic SR/exterior membrane junctions (i.e., dyspedic calcium release units, dCRUs) that contain dihydropyridine receptors (DHPRs) and triadin, two essential components of CRUs, but no RyRs (or feet). Lack of RyRs in turn affects the disposition of DHPRs, which is normally dictated by a linkage to RyR subunits. In the dCRUs of 1B5 cells, DHPRs are neither grouped into tetrads nor aligned in two orthogonal directions. We have explored the structural role of RyR3 in the assembly of CRUs in 1B5 cells independently expressing either RyR1 or RyR3. Either isoform colocalizes with DHPRs and triadin at the cell periphery. Electron microscopy shows that expression of either isoform results in CRUs containing arrays of feet, indicating the ability of both isoforms to be targeted to dCRUs and to assemble in ordered arrays in the absence of the other. However, a significant difference between RyR1- and RyR3-rescued junctions is revealed by freeze fracture. While cells transfected with RyR1 show restoration of DHPR tetrads and DHPR orthogonal alignment indicative of a link to RyRs, those transfected with RyR3 do not. This indicates that RyR3 fails to link to DHPRs in a specific manner. This morphological evidence supports the hypothesis that activation of RyR3 in skeletal muscle cells must be indirect and provides the basis for failure of e-c coupling in muscle cells containing RyR3 but lacking RyR1 (see the accompanying report, ). PMID- 11053126 TI - Divergent functional properties of ryanodine receptor types 1 and 3 expressed in a myogenic cell line. AB - Of the three known ryanodine receptor (RyR) isoforms expressed in muscle, RyR1 and RyR2 have well-defined roles in contraction. However, studies on mammalian RyR3 have been difficult because of low expression levels relative to RyR1 or RyR2. Using the herpes simplex virus 1 (HSV-1) helper-free amplicon system, we expressed either RyR1 or RyR3 in 1B5 RyR-deficient myotubes. Western blot analysis revealed that RyR1- or RyR3-transduced cells expressed the appropriate RyR isoform of the correct molecular mass. Although RyR1 channels exhibited the expected unitary conductance for Cs(+) in bilayer lipid membranes, 74 of 88 RyR3 channels exhibited pronounced subconductance behavior. Western blot analysis with an FKBP12/12.6-selective antibody reveals that differences in gating behavior exhibited by RyR1 and RyR3 may be, in part, the result of lower affinity of RyR3 for FKBP12. In calcium imaging studies, RyR1 restored skeletal-type excitation contraction coupling, whereas RyR3 did not. Although RyR3-expressing myotubes were more sensitive to caffeine than those expressing RyR1, they were much less sensitive to 4-chloro-m-cresol (CMC). In RyR1-expressing cells, regenerative calcium oscillations were observed in response to caffeine and CMC but were never seen in RyR3-expressing 1B5 cells. In [(3)H]ryanodine binding studies, only RyR1 exhibited sensitivity to CMC, but both RyR isoforms responded to caffeine. These functional differences between RyR1 and RyR3 expressed in a mammalian muscle context may reflect differences in association with accessory proteins, especially FKBP12, as well as structural differences in modulator binding sites. PMID- 11053127 TI - Desformylgramicidin: a model channel with an extremely high water permeability. AB - The water conductivity of desformylgramicidin exceeds the permeability of gramicidin A by two orders of magnitude. With respect to its single channel hydraulic permeability coefficient of 1.1.10(-12) cm(3) s(-1), desformylgramicidin may serve as a model for extremely permeable aquaporin water channel proteins (AQP4 and AQPZ). This osmotic permeability exceeds the conductivity that is predicted by the theory of single-file transport. It was derived from the concentration distributions of both pore-impermeable and permeable cations that were simultaneously measured by double barreled microelectrodes in the immediate vicinity of a planar bilayer. From solvent drag experiments, approximately five water molecules were found to be transported by a single-file process along with one ion through the channel. The single channel proton, potassium, and sodium conductivities were determined to be equal to 17 pS (pH 2.5), 7 and 3 pS, respectively. Under any conditions, the desformyl-channel remains at least 10 times longer in its open state than gramicidin A. PMID- 11053128 TI - Two mechanisms of K(+)-dependent potentiation in Kv2.1 potassium channels. AB - Elevation of external [K(+)] potentiates outward K(+) current through several voltage-gated K(+) channels. This increase in current magnitude is paradoxical in that it occurs despite a significant decrease in driving force. We have investigated the mechanisms involved in K(+)-dependent current potentiation in the Kv2.1 K(+) channel. With holding potentials of -120 to -150 mV, which completely removed channels from the voltage-sensitive inactivated state, elevation of external [K(+)] up to 10 mM produced a concentration-dependent increase in outward current magnitude. In the absence of inactivation, currents were maximally potentiated by 38%. At more positive holding potentials, which produced steady-state inactivation, K(+)-dependent potentiation was enhanced. The additional K(+)-dependent potentiation (above 38%) at more positive holding potentials was precisely equal to a K(+)-dependent reduction in steady-state inactivation. Mutation of two lysine residues in the outer vestibule of Kv2.1 (K356 and K382), to smaller, uncharged residues (glycine and valine, respectively), completely abolished K(+)-dependent potentiation that was not associated with inactivation. These mutations did not influence steady-state inactivation or the K(+)-dependent potentiation due to reduction in steady-state inactivation. These results demonstrate that K(+)-dependent potentiation can be completely accounted for by two independent mechanisms: one that involved the outer vestibule lysines and one that involved K(+)-dependent removal of channels from the inactivated state. Previous studies demonstrated that the outer vestibule of Kv2.1 can be in at least two conformations, depending on the occupancy of the selectivity filter by K(+) (Immke, D., M. Wood, L. Kiss, and S. J. Korn. 1999. J. Gen. Physiol. 113:819-836; Immke, D., and S. J. Korn. 2000. J. Gen. Physiol. 115:509-518). This change in conformation was functionally defined by a change in TEA sensitivity. Similar to the K(+)-dependent change in TEA sensitivity, the lysine-dependent potentiation depended primarily (>90%) on Lys 356 and was enhanced by lowering initial K(+) occupancy of the pore. Furthermore, the K(+)-dependent changes in current magnitude and TEA sensitivity were highly correlated. These results suggest that the previously described K(+)-dependent change in outer vestibule conformation underlies the lysine-sensitive, K(+) dependent potentiation mechanism. PMID- 11053129 TI - G(i)-dependent localization of beta(2)-adrenergic receptor signaling to L-type Ca(2+) channels. AB - A plausible determinant of the specificity of receptor signaling is the cellular compartment over which the signal is broadcast. In rat heart, stimulation of beta(1)-adrenergic receptor (beta(1)-AR), coupled to G(s)-protein, or beta(2)-AR, coupled to G(s)- and G(i)-proteins, both increase L-type Ca(2+) current, causing enhanced contractile strength. But only beta(1)-AR stimulation increases the phosphorylation of phospholamban, troponin-I, and C-protein, causing accelerated muscle relaxation and reduced myofilament sensitivity to Ca(2+). beta(2)-AR stimulation does not affect any of these intracellular proteins. We hypothesized that beta(2)-AR signaling might be localized to the cell membrane. Thus we examined the spatial range and characteristics of beta(1)-AR and beta(2)-AR signaling on their common effector, L-type Ca(2+) channels. Using the cell attached patch-clamp technique, we show that stimulation of beta(1)-AR or beta(2) AR in the patch membrane, by adding agonist into patch pipette, both activated the channels in the patch. But when the agonist was applied to the membrane outside the patch pipette, only beta(1)-AR stimulation activated the channels. Thus, beta(1)-AR signaling to the channels is diffusive through cytosol, whereas beta(2)-AR signaling is localized to the cell membrane. Furthermore, activation of G(i) is essential to the localization of beta(2)-AR signaling because in pertussis toxin-treated cells, beta(2)-AR signaling becomes diffusive. Our results suggest that the dual coupling of beta(2)-AR to both G(s)- and G(i) proteins leads to a highly localized beta(2)-AR signaling pathway to modulate sarcolemmal L-type Ca(2+) channels in rat ventricular myocytes. PMID- 11053130 TI - Na(+) transport, and the E(1)P-E(2)P conformational transition of the Na(+)/K(+) ATPase. AB - We have used admittance analysis together with the black lipid membrane technique to analyze electrogenic reactions within the Na(+) branch of the reaction cycle of the Na(+)/K(+)-ATPase. ATP release by flash photolysis of caged ATP induced changes in the admittance of the compound membrane system that are associated with partial reactions of the Na(+)/K(+)-ATPase. Frequency spectra and the Na(+) dependence of the capacitive signal are consistent with an electrogenic or electroneutral E(1)P <--> E(2)P conformational transition which is rate limiting for a faster electrogenic Na(+) dissociation reaction. We determine the relaxation rate of the rate-limiting reaction and the equilibrium constants for both reactions at pH 6.2-8.5. The relaxation rate has a maximum value at pH 7.4 (approximately 320 s(-1)), which drops to acidic (approximately 190 s(-1)) and basic (approximately 110 s(-1)) pH. The E(1)P <--> E(2)P equilibrium is approximately at a midpoint position at pH 6.2 (equilibrium constant approximately 0.8) but moves more to the E(1)P side at basic pH 8.5 (equilibrium constant approximately 0.4). The Na(+) affinity at the extracellular binding site decreases from approximately 900 mM at pH 6.2 to approximately 200 mM at pH 8.5. The results suggest that during Na(+) transport the free energy supplied by the hydrolysis of ATP is mainly used for the generation of a low-affinity extracellular Na(+) discharge site. Ionic strength and lyotropic anions both decrease the relaxation rate. However, while ionic strength does not change the position of the conformational equilibrium E(1)P <--> E(2)P, lyotropic anions shift it to E(1)P. PMID- 11053131 TI - Mechanism generating endocochlear potential: role played by intermediate cells in stria vascularis. AB - The endocochlear DC potential (EP) is generated by the stria vascularis, and essential for the normal function of hair cells. Intermediate cells are melanocytes in the stria vascularis. To examine the contribution of the membrane potential of intermediate cells (E(m)) to the EP, a comparison was made between the effects of K(+) channel blockers on the E(m) and those on the EP. The E(m) of dissociated guinea pig intermediate cells was measured in the zero-current clamp mode of the whole-cell patch clamp configuration. The E(m) changed by 55.1 mV per 10-fold changes in extracellular K(+) concentration. Ba(2+), Cs(+), and quinine depressed the E(m) in a dose-dependent manner, whereas tetraethylammonium at 30 mM and 4-aminopyridine at 10 mM had no effect. The reduction of the E(m) by Ba(2+) and Cs(+) was enhanced by lowering the extracellular K(+) concentration from 3.6 mM to 1.2 mM. To examine the effect of the K(+) channel blockers on the EP, the EP of guinea pigs was maintained by vascular perfusion, and K(+) channel blockers were administered to the artificial blood. Ba(2+), Cs(+) and quinine depressed the EP in a dose-dependent manner, whereas tetraethylammonium at 30 mM and 4-aminopyridine at 10 mM did not change the EP. A 10-fold increase in the K(+) concentration in the artificial blood caused a minor decrease in the EP of only 10.6 mV. The changes in the EP were similar to those seen in the E(m) obtained at the lower extracellular K(+) concentration of 1.2 mM. On the basis of these results, we propose that the EP is critically dependent on the voltage jump across the plasma membrane of intermediate cells, and that K(+) concentration in the intercellular space in the stria vascularis may be actively controlled at a concentration lower than the plasma level. PMID- 11053132 TI - Inclusion-induced bilayer deformations: effects of monolayer equilibrium curvature. AB - The energetics of protein-induced bilayer deformation in systems with finite monolayer equilibrium curvature were investigated using an elastic membrane model. In this model the bilayer deformation energy delta G(def) has two major components: a compression-expansion component and a splay-distortion component, which includes the consequences of a bilayer curvature frustration due to a monolayer equilibrium curvature, c(0), that is different from zero. For any choice of bilayer material constants, the value of delta G(def) depends on global bilayer properties, as described by the bilayer material constants, as well as the energetics of local lipid packing adjacent to the protein. We introduce this dependence on lipid packing through the contact slope, s, at the protein-bilayer boundary. When c(0) = 0, delta G(def) can be approximated as a biquadratic function of s and the monolayer deformation at the protein/bilayer boundary, u(0): delta G(def) = a(1)u(0)(2) + a(2)u(0)s + a(3)s(2), where a(1), a(2), and a(3) are functions of the bilayer thickness, the bilayer compression-expansion and splay-distortion moduli, and the inclusion radius (this expression becomes exact when the Gaussian curvature component of delta G(def) is negligible). When c(0) not equal 0, the curvature frustration contribution is determined by the choice of boundary conditions at the protein-lipid boundary (by the value of s), and delta G(def) is the sum of the energy for c(0) = 0 plus the curvature frustration-dependent contribution. When the energetic penalty for the local lipid packing can be ignored, delta G(def) will be determined only by the global bilayer properties, and a c(0) > 0 will tend to promote a local inclusion-induced bilayer thinning. When the energetic penalty for local lipid packing is large, s will be constrained by the value of c(0). In a limiting case, where s is determined only by geometric constraints imposed by c(0), a c(0) > 0 will impede such local bilayer thinning. One cannot predict curvature effects without addressing the proper choice of boundary conditions at the protein-bilayer contact surface. PMID- 11053133 TI - Domain growth, shapes, and topology in cationic lipid bilayers on mica by fluorescence and atomic force microscopy. AB - Domain formation in mica-supported cationic bilayers of dipalmitoyltrimethylammoniumpropane (DPTAP) and dimyristoyltrimethylammoniumpropane (DMTAP), fluorescently doped with an NBD (((7 nitro-2-1, 3-benzoxadiazol-4-yl)amino)caproyl) phospholipid, was investigated with fluorescence microscopy and atomic force microscopy. Heating above the acyl chain melting temperature and cooling to room temperature resulted in nucleation and growth of domains with distinguishable patterns. Fractal patterns were found for DPTAP, whereas DMTAP domains were elongated and triangular with feathery edges. Reducing the cooling rate or probe concentration for DPTAP bilayers resulted in larger, filled-in domains with more rounded edges. However, for DMTAP, cooling rates mainly affected size and only slightly modified domain morphology. In a saline environment, the domains were dark, and the surrounding continuous region was bright and thus contained the fluorescent probe. However, as the salt concentration was decreased, the dark regions percolated (connected), resulting in bright domains. Atomic force microscopy scans along domain edges revealed that the dark regions in fluorescence images were approximately 1.4 nm thicker than the light regions. Additionally, the dark regions were of bilayer thickness, approximately 4 nm. Comparison of these results in bilayers to well documented behavior in Langmuir monolayers has revealed many similarities (and some differences) and is therefore useful for understanding our observations and identifying possible growth mechanisms that may occur in domain formation in cell membranes or supported membrane systems. PMID- 11053134 TI - The structure of D-erythro-C18 ceramide at the air-water interface. AB - X-ray reflectivity (XR) and diffraction at grazing angles of incidence (GID) were conducted to determine the structure of synthetic D-erythro C18-ceramide films at the air-water interface at various surface pressures (pi). Analysis of the GID reveals that the monomolecular film, at the crystalline phase (pi > 0 mN/m), is predominantly hexagonal. In this crystalline phase, the analysis of the reflectivity yields an electron density profile that consists of three distinct homogeneous slabs, one associated with the headgroup region and the other two with the hydrocarbon chains. At large molecular areas (pi approximately 0), isolated crystalline domains coexist with two-dimensional gas phase. Within the crystalline domains, we find an orthorhombic arrangement of the chains that coexists with the hexagonal symmetry. It is argued that the two-dimensional orthorhombic crystals are induced by hydrogen bonding between headgroups even at very low surface pressures. Although their structure is incommensurate with the simple hexagonal arrangement, they act as nucleation centers for the conventional hexagonal phase which dominates at high pi. PMID- 11053135 TI - PMP1 18-38, a yeast plasma membrane protein fragment, binds phosphatidylserine from bilayer mixtures with phosphatidylcholine: a (2)H-NMR study. AB - PMP1 is a 38-residue plasma membrane protein of the yeast Saccharomyces cerevisiae that regulates the activity of the H(+)-ATPase. The cytoplasmic domain conformation results in a specific interfacial distribution of five basic side chains, thought to strongly interact with anionic phospholipids. We have used the PMP1 18-38 fragment to carry out a deuterium nuclear magnetic resonance ((2)H NMR) study for investigating the interactions between the PMP1 cytoplasmic domain and phosphatidylserines. For this purpose, mixed bilayers of 1-palmitoyl, 2 oleoyl-sn-glycero-3-phosphocholine (POPC) and 1-palmitoyl, 2-oleoyl-sn-glycero-3 phosphoserine (POPS) were used as model membranes (POPC/POPS 5:1, m/m). Spectra of headgroup- and chain-deuterated POPC and POPS phospholipids, POPC-d4, POPC d31, POPS-d3, and POPS-d31, were recorded at different temperatures and for various concentrations of the PMP1 fragment. Data obtained from POPS deuterons revealed the formation of specific peptide-POPS complexes giving rise to a slow exchange between free and bound PS lipids, scarcely observed in solid-state NMR studies of lipid-peptide/protein interactions. The stoichiometry of the complex (8 POPS per peptide) was determined and its significance is discussed. The data obtained with headgroup-deuterated POPC were rationalized with a model that integrates the electrostatic perturbation induced by the cationic peptide on the negatively charged membrane interface, and a "spacer" effect due to the intercalation of POPS/PMP1f complexes between choline headgroups. PMID- 11053136 TI - Cholesterol dependent recruitment of di22:6-PC by a G protein-coupled receptor into lateral domains. AB - Bovine rhodopsin was reconstituted into mixtures of didocosahexaenoylphosphatidylcholine (di22:6-PC), dipalmitoylphosphatidylcholine (di16:0-PC), sn-1-palmitoyl-sn-2-docosahexaenoylphosphatidylcholine (16:0, 22:6 PC) and cholesterol. Rhodopsin denaturation was examined by using high sensitivity differential scanning calorimetry. The unfolding temperature was increased at lower levels of lipid unsaturation, but the highest temperature was detected for native disk membranes: di22:6-PC < 16:0,22:6-PC < di16:0,18:1-PC < native disks. The incorporation of 30 mol% of cholesterol resulted in 2-4 degrees C increase of denaturation temperature in all reconstituted systems examined. From the analysis of van't Hoff's and calorimetric enthalpies, it was concluded that the presence of cholesterol in di22:6-PC-containing bilayers induces a level of cooperativity in rhodopsin unfolding. Fluorescence resonance energy transfer (FRET), using lipids labeled at the headgroup with pyrene (Py) as donors and rhodopsin retinal group as acceptor of fluorescence, was used to study rhodopsin association with lipids. Higher FRET efficiencies detected for di22:6-PE-Py, compared to di16:0-PE-Py, in mixed di22:6-PC-di16:0-PC-cholesterol bilayers, indicate preferential segregation of rhodopsin with polyunsaturated lipids. The effective range of the rhodopsin-lipid interactions facilitating cluster formation exceeds two adjacent lipid layers. In similar mixed bilayers containing no cholesterol, cluster formation was absent at temperatures above lipid phase transition, indicating a crucial role of cholesterol in microdomain formation. PMID- 11053137 TI - The topology of lysine-containing amphipathic peptides in bilayers by circular dichroism, solid-state NMR, and molecular modeling. AB - In order to better understand the driving forces that determine the alignment of amphipathic helical polypeptides with respect to the surface of phospholipid bilayers, lysine-containing peptide sequences were designed, prepared by solid phase chemical synthesis, and reconstituted into membranes. CD spectroscopy indicates that all peptides exhibit a high degree of helicity in the presence of SDS micelles or POPC small unilamellar vesicles. Proton-decoupled (31)P-NMR solid state NMR spectroscopy demonstrates that in the presence of peptides liquid crystalline phosphatidylcholine membranes orient well along glass surfaces. The orientational distribution and dynamics of peptides labeled with (15)N at selected sites were investigated by proton-decoupled (15)N solid-state NMR spectroscopy. Polypeptides with a single lysine residue adopt a transmembrane orientation, thereby locating this polar amino acid within the core region of the bilayer. In contrast, peptides with > or = 3 lysines reside along the surface of the membrane. With 2 lysines in the center of an otherwise hydrophobic amino acid sequence the peptides assume a broad orientational distribution. The energy of lysine discharge, hydrophobic, polar, and all other interactions are estimated to quantitatively describe the polypeptide topologies observed. Furthermore, a molecular modeling algorithm based on the hydrophobicities of atoms in a continuous hydrophilic-hydrophobic-hydrophilic potential describes the experimentally observed peptide topologies well. PMID- 11053138 TI - Pulmonary surfactant protein A interacts with gel-like regions in monolayers of pulmonary surfactant lipid extract. AB - Epifluorescence microscopy was used to investigate the interaction of pulmonary surfactant protein A (SP-A) with spread monolayers of porcine surfactant lipid extract (PSLE) containing 1 mol % fluorescent probe (NBD-PC) spread on a saline subphase (145 mM NaCl, 5 mM Tris-HCl, pH 6.9) containing 0, 0.13, or 0.16 microg/ml SP-A and 0, 1.64, or 5 mM CaCl(2). In the absence of SP-A, no differences were noted in PSLE monolayers in the absence or presence of Ca(2+). Circular probe-excluded (dark) domains were observed against a fluorescent background at low surface pressures (pi approximately 5 mN/m) and the domains grew in size with increasing pi. Above 25 mN/m, the domain size decreased with increasing pi. The amount of observable dark phase was maximal at 18% of the total film area at pi approximately 25 mN/m, then decreased to approximately 3% at pi approximately 40 mN/m. The addition of 0.16 microg/ml SP-A with 0 or 1.64 mM Ca(2+) in the subphase caused an aggregation of dark domains into a loose network, and the total amount of dark phase was increased to approximately 25% between pi of 10-28 mN/m. Monolayer features in the presence of 5 mM Ca(2+) and SP-A were not substantially different from those spread in the absence of SP-A, likely due to a self-association and aggregation of SP-A in the presence of higher concentrations of Ca(2+). PSLE films were spread on a subphase containing 0.16 microg/ml SP-A with covalently bound Texas Red (TR-SP-A). In the absence of Ca(2+), TR-SP-A associated with the reorganized dark phase (as seen with the lipid probe). The presence of 5 mM Ca(2+) resulted in an appearance of TR-SP-A in the fluid phase and of aggregates at the fluid/gel phase boundaries of the monolayers. This study suggests that SP-A associates with PSLE monolayers, particularly with condensed or solid phase lipid, and results in some reorganization of rigid phase lipid in surfactant monolayers. PMID- 11053139 TI - Perturbed equilibria of myosin binding in airway smooth muscle: bond-length distributions, mechanics, and ATP metabolism. AB - We carried out a detailed mathematical analysis of the effects of length fluctuations on the dynamically evolving cross-bridge distributions, simulating those that occur in airway smooth muscle during breathing. We used the latch regulation scheme of Hai and Murphy (Am. J. Physiol. Cell Physiol. 255:C86-C94, 1988) integrated with Huxley's sliding filament theory of muscle contraction. This analysis showed that imposed length fluctuations decrease the mean number of attached bridges, depress muscle force and stiffness, and increase force-length hysteresis. At frequencies >0.1 Hz, the bond-length distribution of slowly cycling latch bridges changed little over the stretch cycle and contributed almost elastically to muscle force, but the rapidly cycling cross-bridge distribution changed substantially and dominated the hysteresis. By contrast, at frequencies <0.033 Hz this behavior was reversed: the rapid cycling cross-bridge distribution changed little, effectively functioning as a constant force generator, while the latch bridge bond distribution changed substantially and dominated the stiffness and hysteresis. The analysis showed the dissociation of force/length hysteresis and cross-bridge cycling rates when strain amplitude exceeds 3%; that is, there is only a weak coupling between net external mechanical work and the ATP consumption required for cycling cross-bridges during the oscillatory steady state. Although these results are specific to airway smooth muscle, the approach generalizes to other smooth muscles subjected to cyclic length fluctuations. PMID- 11053140 TI - Distribution of proteins implicated in excitation-contraction coupling in rat ventricular myocytes. AB - We have examined the distribution of ryanodine receptors, L-type Ca(2+) channels, calsequestrin, Na(+)/Ca(2+) exchangers, and voltage-gated Na(+) channels in adult rat ventricular myocytes. Enzymatically dissociated cells were fixed and dual labeled with specific antibodies using standard immunocytochemistry protocols. Images were deconvolved to reverse the optical distortion produced by wide-field microscopes equipped with high numerical aperture objectives. Every image showed a well-ordered array of fluorescent spots, indicating that all of the proteins examined were distributed in discrete clusters throughout the cell. Mathematical analysis of the images revealed that dyads contained only ryanodine receptors, L type Ca(2+) channels, and calsequestrin, and excluded Na(+)/Ca(2+) exchangers and voltage-gated Na(+) channels. The Na(+)/Ca(2+) exchanger and voltage-gated Na(+) channels were distributed largely within the t-tubules, on both transverse and axial elements, but were not co-localized. The t-tubule can therefore be subdivided into at least three structural domains; one of coupling (dyads), one containing the Na(+)/Ca(2+) exchanger, and one containing voltage-gated Na(+) channels. We conclude that if either the slip mode conductance of the Na(+) channel or the reverse mode of the Na(+)/Ca(2+) exchanger are to contribute to the contractile force, the fuzzy space must extend outside of the dyad. PMID- 11053141 TI - Multimerization-cyclization of DNA fragments as a method of conformational analysis. AB - Ligation of short DNA fragments results in the formation of linear and circular multimers of various lengths. The distribution of products in such a reaction is often used to evaluate fragment bending caused by specific chemical modification, by bound ligands or by the presence of irregular structural elements. We have developed a more rigorous quantitative approach to the analysis of such experimental data based on determination of j-factors for different multimers from the distribution of the reaction products. j-Factors define the effective concentration of one end of a linear chain in the vicinity of the other end. To extract j-factors we assumed that kinetics of the reaction is described by a system of differential equations where j-factors appear as coefficients. The assumption was confirmed by comparison with experimental data obtained here for DNA fragments containing A-tracts. At the second step of the analysis j-factors are used to determine conformational parameters of DNA fragments: the equilibrium bend angle, the bending rigidity of the fragment axis, and the total twist of the fragments. This procedure is based on empirical equations that connect the conformational parameters with the set of j-factors. To obtain the equations, we computed j-factors for a large array of conformational parameters that describe model fragments. The approach was tested on both simulated and actual experimental data for DNA fragments containing A-tracts. A-tract DNA bend angle determined here is in good agreement with previously published data. We have established a set of experimental conditions necessary for the data analysis to be successful. PMID- 11053142 TI - Characterization of the proton-transporting photocycle of pharaonis halorhodopsin. AB - The photocycle of pharaonis halorhodopsin was investigated in the presence of 100 mM NaN(3) and 1 M Na(2)SO(4). Recent observations established that the replacement of the chloride ion with azide transforms the photocycle from a chloride-transporting one into a proton-transporting one. Kinetic analysis proves that the photocycle is very similar to that of bacteriorhodopsin. After K and L, intermediate M appears, which is missing from the chloride-transporting photocycle. In this intermediate the retinal Schiff base deprotonates. The rise of M in halorhodopsin is in the microsecond range, but occurs later than in bacteriorhodopsin, and its decay is more accentuated multiphasic. Intermediate N cannot be detected, but a large amount of O accumulates. The multiphasic character of the last step of the photocycle could be explained by the existence of a HR' state, as in the chloride photocycle. Upon replacement of chloride ion with azide, the fast electric signal changes its sign from positive to negative, and becomes similar to that detected in bacteriorhodopsin. The photocycle is enthalpy-driven, as is the chloride photocycle of halorhodopsin. These observations suggest that, while the basic charge translocation steps become identical to those in bacteriorhodopsin, the storage and utilization of energy during the photocycle remains unchanged by exchanging chloride with azide. PMID- 11053143 TI - Kinetics of local helix formation in poly-L-glutamic acid studied by time resolved photoacoustics: neutralization reactions of carboxylates in aqueous solutions and their relevance to the problem of protein folding. AB - Photoactivatable caged protons have been used to trigger proton transfer reactions in aqueous solutions of acetate, glutamate, and poly-L-glutamic acid, and the volumetric and enthalpic changes have been detected and characterized by means of time-resolved photoacoustics. Neutralization of carboxylates in aqueous solutions invariably results in an expansion of the solution due to the disappearance of two charges and is accompanied by little enthalpic change. The reactions occur with thermally activated, apparent bimolecular rates on the order of 10(10) M(-1)s(-1). In the case of aqueous solutions of poly-L-glutamic acid at pH around the pK(a) of the coil-to-helix transition, diffusional binding of a proton by carboxylates is followed by a sequential reaction with rate 1.06 (+/- 0.05) x 10(7)s(-1). This step is not thermally activated in the temperature range we have investigated and is likely related to local formation of hydrogen bonds near the protonation site. This structural event may constitute a rate-limiting step in helix propagation. PMID- 11053144 TI - Pathways in two-state protein folding. AB - Thermodynamic measurements of proteins indicate that the folding to the native state takes place either through stable intermediates or through a two-state process without intermediates. The rather short folding times of proteins indicate that folding is guided through some sequence of contact bindings. We discuss the possibility of reconciling a two-state folding event with a sequential folding process in a schematic model of protein folding. We propose a new dynamical transition temperature that is lower than the temperature at which proteins in equilibrium unfold. This is in qualitative agreement with observations of in vivo protein folding activity quantified by chaperone concentration in Escherichia coli. Finally, we discuss our framework in connection with the unfolding of proteins at low temperatures. PMID- 11053145 TI - Molecular dynamics of solid-state lysozyme as affected by glycerol and water: a neutron scattering study. AB - Glycerol has been shown to lower the heat denaturation temperature (T(m)) of dehydrated lysozyme while elevating the T(m) of hydrated lysozyme (. J. Pharm. Sci. 84:707-712). Here, we report an in situ elastic neutron scattering study of the effect of glycerol and hydration on the internal dynamics of lysozyme powder. Anharmonic motions associated with structural relaxation processes were not detected for dehydrated lysozyme in the temperature range of 40 to 450K. Dehydrated lysozyme was found to have the highest T(m) by. Upon the addition of glycerol or water, anharmonicity was recovered above a dynamic transition temperature (T(d)), which may contribute to the reduction of T(m) values for dehydrated lysozyme in the presence of glycerol. The greatest degree of anharmonicity, as well as the lowest T(d), was observed for lysozyme solvated with water. Hydrated lysozyme was also found to have the lowest T(m) by. In the regime above T(d), larger amounts of glycerol lead to a higher rate of change in anharmonic motions as a function of temperature, rendering the material more heat labile. Below T(d), where harmonic motions dominate, the addition of glycerol resulted in a lower amplitude of motions, correlating with a stabilizing effect of glycerol on the protein. PMID- 11053146 TI - A flash photolysis method to characterize hexacoordinate hemoglobin kinetics. AB - A flash photolysis method is described for analyzing ligand binding to the new and growing group of hemoglobins which are hexacoordinate in the unligated, ferrous state. Simple analysis of a two exponential fit to time courses for CO rebinding at varying CO concentrations yields rate constants for formation and dissociation of the hexacoordinate complex, and the bimolecular rate constant for CO binding. This method was tested with a nonsymbiotic plant hemoglobin from rice for which these values had not previously been determined. For this protein, dissociation and rebinding of the hexacoordinating amino acid side chain, His(73), is rapid and similar to the rate of CO binding at high CO concentrations. These results indicate that hexacoordination must be taken into account when evaluating the affinity of hexacoordinate hemoglobins for ligands. PMID- 11053147 TI - A systematic study of the vibrational free energies of polypeptides in folded and random states. AB - Molecular vibrations, especially low frequency motions, may be used as an indication of the rigidity or the flatness of the protein folding energy landscape. We have studied the vibrational properties of native folded as well as random coil structures of more than 60 polypeptides. The picture we obtain allows us to perceive how and why the energy landscape progressively rigidifies while still allowing potential flexibility. Compared with random coil structures, both alpha-helices and beta-hairpins are vibrationally more flexible. The vibrational properties of loop structures are similar to those of the corresponding random coil structures. Inclusion of an alpha-helix tends to rigidify peptides and so called building blocks of the structure, whereas the addition of a beta-structure has less effect. When small building blocks coalesce to form larger domains, the protein rigidifies. However, some folded native conformations are still found to be vibrationally more flexible than random coil structures, for example, beta(2) microglobulin and the SH3 domain. Vibrational free energy contributes significantly to the thermodynamics of protein folding and affects the distribution of the conformational substates. We found a weak correlation between the vibrational folding energy and the protein size, consistent with both previous experimental estimates and theoretical partition of the heat capacity change in protein folding. PMID- 11053148 TI - Serine and threonine residues bend alpha-helices in the chi(1) = g(-) conformation. AB - The relationship between the Ser, Thr, and Cys side-chain conformation (chi(1) = g(-), t, g(+)) and the main-chain conformation (phi and psi angles) has been studied in a selection of protein structures that contain alpha-helices. The statistical results show that the g(-) conformation of both Ser and Thr residues decreases their phi angles and increases their psi angles relative to Ala, used as a control. The additional hydrogen bond formed between the O(gamma) atom of Ser and Thr and the i-3 or i-4 peptide carbonyl oxygen induces or stabilizes a bending angle in the helix 3-4 degrees larger than for Ala. This is of particular significance for membrane proteins. Incorporation of this small bending angle in the transmembrane alpha-helix at one side of the cell membrane results in a significant displacement of the residues located at the other side of the membrane. We hypothesize that local alterations of the rotamer configurations of these Ser and Thr residues may result in significant conformational changes across transmembrane helices, and thus participate in the molecular mechanisms underlying transmembrane signaling. This finding has provided the structural basis to understand the experimentally observed influence of Ser residues on the conformational equilibrium between inactive and active states of the receptor, in the neurotransmitter subfamily of G protein-coupled receptors. PMID- 11053149 TI - The influence of water on the nanomechanical behavior of the plant biopolyester cutin as studied by AFM and solid-state NMR. AB - Atomic force microscopy and solid-state nuclear magnetic resonance have been used to investigate the effect of water absorption on the nanoscale elastic properties of the biopolyester, cutin, isolated from tomato fruit cuticle. Changes in the humidity and temperature at which fruits are grown or stored can affect the plant surface (cuticle) and modify its susceptibility to pathogenic attack by altering the cuticle's rheological properties. In this work, atomic force microscopy measurements of the surface mechanical properties of isolated plant cutin have been made as a first step to probing the impact of water uptake from the environment on surface flexibility. A dramatic decrease in surface elastic modulus (from approximately 32 to approximately 6 MPa) accompanies increases in water content as small as 2 wt %. Complementary solid-state nuclear magnetic resonance measurements reveal enhanced local mobility of the acyl chain segments with increasing water content, even at molecular sites remote from the covalent cross-links that are likely to play a crucial role in cutin's elastic properties. PMID- 11053150 TI - Distribution of ganglioside GM1 in L-alpha dipalmitoylphosphatidylcholine/cholesterol monolayers: a model for lipid rafts. AB - The distribution of low concentrations of ganglioside GM1 in L-alpha dipalmitoylphosphatidylcholine (DPPC) and DPPC/cholesterol monolayers supported on mica has been studied using atomic force microscopy (AFM). The monolayers studied correspond to a pure gel phase and a mixture of liquid-expanded (LE) and liquid-condensed (LC) phases for DPPC and to a single homogeneous liquid-ordered phase for 2:1 DPPC/cholesterol. The addition of 2.5-5% GM1 to phase-separated DPPC monolayers resulted in small round ganglioside-rich microdomains in the center and at the edges of the LC domains. Higher amounts of GM1 (10%) give numerous filaments in the center of the LC domains and larger patches at the edges. A gel phase DPPC monolayer containing GM1 showed large domains containing a network of GM1-rich filaments. The addition of GM1 to a liquid-ordered 2:1 DPPC/cholesterol monolayer gives small, round domains that vary in size from 50 to 150 nm for a range of surface pressures. Larger amounts of GM1 lead to coalescence of the small, round domains to give longer filaments that cover 30 40% of the monolayer surface for 10 mol % GM1. The results indicate that biologically relevant GM1 concentrations lead to submicron-sized domains in a cholesterol-rich liquid-ordered phase that is analogous to that found in detergent-insoluble membrane fractions, and are thought to be important in membrane microdomains or rafts. This demonstrates that AFM studies of model monolayers and bilayers provide a powerful method for the direct detection of microdomains that are too small for study with most other techniques. PMID- 11053151 TI - Using patient-identifiable data for observational research and audit. PMID- 11053152 TI - The current status of psychological debriefing. PMID- 11053153 TI - Driving after repair of groin hernia. PMID- 11053154 TI - Time to talk about rape. PMID- 11053155 TI - Bicycle helmets: it's time to use them. PMID- 11053156 TI - Ebola virus claims more lives in Uganda. PMID- 11053157 TI - Phenylpropanolamine in drugs could be a risk for stroke. PMID- 11053158 TI - "Labour" think tank proposes privatisation of clinical services. PMID- 11053159 TI - VSO launches campaign to increase access to AIDS drugs. PMID- 11053160 TI - In brief PMID- 11053161 TI - Biotechnology company's shares dive 67% after drug failure. PMID- 11053163 TI - Statins may reduce risk of Alzheimer's disease PMID- 11053162 TI - No advantage in screening for endometrial cancer PMID- 11053164 TI - "Designer baby" cures sister. PMID- 11053165 TI - Greenpeace acts on life threatening poisons in Gujara. PMID- 11053166 TI - Researchers able to check chromosome numbers in embryos PMID- 11053167 TI - Early exposure to cows' milk raises risk of diabetes in high risk children PMID- 11053168 TI - Transplanted cells revive heart muscle. PMID- 11053169 TI - Genetically engineered sweetcorn may be in US food chain. PMID- 11053170 TI - Abortion pill withdrawn in germany after financial losses. PMID- 11053171 TI - UK is losing market share in pharmaceutical research. PMID- 11053172 TI - Of pills and ills. PMID- 11053173 TI - Randomised controlled trial of midwife led debriefing to reduce maternal depression after operative childbirth. AB - OBJECTIVE: To assess the effectiveness of a midwife led debriefing session during the postpartum hospital stay in reducing the prevalence of maternal depression at six months postpartum among women giving birth by caesarean section, forceps, or vacuum extraction. DESIGN: Randomised controlled trial. SETTING: Large maternity teaching hospital in Melbourne, Australia. PARTICIPANTS: 1041 women who had given birth by caesarean section (n= 624) or with the use of forceps (n= 353) or vacuum extraction (n= 64). MAIN OUTCOME MEASURES: Maternal depression (score >/=13 on the Edinburgh postnatal depression scale) and overall health status (comparison of mean scores on SF-36 subscales) measured by postal questionnaire at six months postpartum. RESULTS: 917 (88%) of the women recruited responded to the outcome questionnaire. More women allocated to debriefing scored as depressed six months after birth than women allocated to usual postpartum care (81 (17%) v 65 (14%)), although this difference was not significant (odds ratio=1.24, 95% confidence interval 0.87 to 1.77). They were also more likely to report that depression had been a problem for them since the birth, but the difference was not significant (123 (28%) v 94 (22%); odds ratio=1. 37, 1.00 to 1.86). Women allocated to debriefing had poorer health status on seven of the eight SF-36 subscales, although the difference was significant only for role functioning (emotional): mean scores 73.32 v 78.98, t= -2.31, 95% confidence interval -10.48 to -0.84). CONCLUSIONS: Midwife led debriefing after operative birth is ineffective in reducing maternal morbidity at six months postpartum. The possibility that debriefing contributed to emotional health problems for some women cannot be excluded. PMID- 11053174 TI - Randomised comparison of the effectiveness and costs of community and hospital based mental health services for children with behavioural disorders. AB - OBJECTIVE: To test the hypothesis that a community based intervention by secondary child and adolescent mental health services would be significantly more effective and less costly than a hospital based intervention. DESIGN: Open study with two randomised parallel groups. SETTING: Two health districts in the north of England. PARTICIPANTS: Parents of 3 to 10 year old children with behavioural disorder who had been referred to child and adolescent mental health services. INTERVENTION: Parental education groups. MAIN OUTCOME MEASURES: Parents' and teachers' reports of the child's behaviour, parental depression, parental criticism of the child, impact of the child's behaviour on the family. RESULTS: 141 subjects were randomised to community (n=72) or hospital (n=69) treatment. Primary outcome data were obtained on 115 (82%) cases a year later. Intention to treat analyses showed no significant differences between the community and hospital based groups on any of the outcome measures, or on costs. Parental depression was common and predicted the child's outcome. CONCLUSIONS: Location of child mental health services may be less important than the range of services that they provide, which should include effective treatment for parents' mental health problems. PMID- 11053176 TI - The doctor is on your side PMID- 11053175 TI - Qualitative analysis of stroke patients' motivation for rehabilitation. AB - OBJECTIVE: To explore the attitudes and beliefs of stroke patients identified by professionals as having either "high" or "low" motivation for rehabilitation. DESIGN: Qualitative study with semistructured interviews. SETTING: The stroke unit of an inner city teaching hospital. PARTICIPANTS: 22 patients with stroke who were undergoing rehabilitation; 14 with high motivation for rehabilitation and eight with low motivation. RESULTS: All patients thought rehabilitation was important for recovery. High motivation patients were more likely to view rehabilitation as the most important means of recovery and to accord themselves an active role in rehabilitation. These patients were also more likely to understand rehabilitation and in particular to understand the specialist role of the nursing staff. Many patients reported independence at home as a personal goal, though few low motivation patients related this goal to success in rehabilitation. Information from professionals about rehabilitation, favourable comparisons with other stroke patients, and the desire to leave hospital had a positive effect on motivation. Conversely, overprotection from family members and professionals, lack of information or the receipt of "mixed messages" from professionals, and unfavourable comparisons with other patients had a negative effect. CONCLUSIONS: There are some differences in beliefs between stroke patients identified as having low or high motivation for rehabilitation. These beliefs seem to be influenced by the environment in which the patient is rehabilitated. Professionals and carers should be made aware of the ways in which their behaviour can positively and negatively affect motivation. PMID- 11053177 TI - Trends in serious head injuries among cyclists in England: analysis of routinely collected data. PMID- 11053178 TI - Advice on driving after groin hernia surgery in the United Kingdom: questionnaire survey. PMID- 11053179 TI - Food for thought PMID- 11053180 TI - Cross sectional study of primary care groups in London: association of measures of socioeconomic and health status with hospital admission rates. AB - OBJECTIVES: To calculate socioeconomic and health status measures for the primary care groups in London and to examine the association between these measures and hospital admission rates. DESIGN: Cross sectional study. SETTING: 66 primary care groups in London, total list size 8.0 million people. MAIN OUTCOME MEASURES: Elective and emergency standardised hospital admission ratios; standardised admission rates for diabetes and asthma. RESULTS: Standardised hospital admission ratios varied from 74 to 116 for total admissions and from 50 to 124 for emergency admissions. Directly standardised admission rates for asthma varied from 152 to 801 per 100 000 (mean 364) and for diabetes from 235 to 1034 per 100 000 (mean 538). There were large differences in the mortality, socioeconomic, and general practice characteristics of the primary care groups. Hospital admission rates were significantly correlated with many of the measures of chronic illness and deprivation. The strongest correlations were with disability living allowance (R=0.64 for total admissions and R=0.62 for emergency admissions, P<0.0001). Practice characteristics were less strongly associated with hospital admission rates. CONCLUSIONS: It is feasible to produce a range of socioeconomic, health status, and practice measures for primary care groups for use in needs assessment and in planning and monitoring health services. These measures show that primary care groups have highly variable patient and practice characteristics and that hospital admission rates are associated with chronic illness and deprivation. These variations will need to be taken into account when assessing performance. PMID- 11053181 TI - Science, medicine, and the future: susceptibility to infection. PMID- 11053182 TI - Lesson of the week: cholesterol emboli syndrome. PMID- 11053183 TI - On the receiving end PMID- 11053184 TI - ABC of colorectal cancer: the role of primary care. PMID- 11053186 TI - Why journals should not publish articles funded by the tobacco industry. PMID- 11053185 TI - Is clinical breast examination an acceptable alternative to mammographic screening? PMID- 11053187 TI - Why do patients need info? PMID- 11053188 TI - Obituaries PMID- 11053189 TI - Impact of NHS direct on demand for immediate care. Target communities show poor awareness of NHS direct. PMID- 11053190 TI - Impact of NHS direct on demand for immediate care. Service has not decreased attendance at one paediatric A and E department. PMID- 11053191 TI - Evidence on endometriosis. Elitism about randomised controlled trials is inappropriate. PMID- 11053192 TI - Seeing what you want to see in randomised controlled trials. Authors' choice of study was ill informed. PMID- 11053193 TI - Seeing what you want to see in randomised controlled trials. Meta- analyses may suffer from interpretation bias too. PMID- 11053194 TI - Thyroid function tests. Tests must still be done in possible thyroid dysfunction. PMID- 11053195 TI - Thyroid function tests. Thyroid stimulating hormone outside the normal range has important implications. PMID- 11053196 TI - Thyroid function tests. Accurate diagnosis depends on both clinical judgment and results of tests. PMID- 11053197 TI - Thyroid function tests. Doing more tests is not always better. PMID- 11053198 TI - Thyroid function tests. Thyroid function testing means different things to different people. PMID- 11053199 TI - Incidence and remission of lower urinary tract symptoms. Authors should have used standardised questionnaire. PMID- 11053200 TI - DoH explains thinking behind national service framework for coronary heart disease. PMID- 11053201 TI - Long standing heart disease should be better screened. PMID- 11053203 TI - Advice on driving after hernia surgery is inconsistent PMID- 11053202 TI - Discontinuation rates for use of statins are high. PMID- 11053204 TI - Reducing stress among junior doctors PMID- 11053205 TI - Inhibition of neuroeffector transmission in human vas deferens by sildenafil. AB - Sildenafil (0.1 - 30 microM), a cyclic GMP phosphodiesterase 5 (PDE 5) inhibitor, induced inhibition of electrically evoked contractions of ring segments of human vas deferens from 34 vasectomies. Zaprinast (0.1 - 100 microM), another PDE 5 inhibitor, and the nitric oxide (NO) donor sodium nitroprusside (SNP) (0.1 - 100 microM) had no effect on neurogenic contractions. The inhibition induced by sildenafil was not modified by the inhibitor of guanylate cyclase 1H [1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ) (1 - 30 microM) but it was abolished by the K(+) channel blockers tetraethylammonium (TEA, 1 mM), iberiotoxin (0.1 microM) and charybdotoxin (0.1 microM). Sildenafil, zaprinast and SNP did not affect the contractions induced by noradrenaline. SNP (10 microM) caused elevation of cyclic GMP levels that was potentiated by sildenafil (10 microM) and zaprinast (100 microM). ODQ (10 microM) inhibited the increase in cyclic GMP. Sildenafil inhibits adrenergic neurotransmission in human vas deferens. The inhibition is not related to accumulation of cyclic GMP but is probably due to activation of prejunctional large-conductance Ca(2+)-activated K(+) channels. PMID- 11053206 TI - Blockade and reversal of spinal morphine tolerance by peptide and non-peptide calcitonin gene-related peptide receptor antagonists. AB - This study examined the effects of the peptide CGRP receptor antagonist CGRP(8 37) and the newly-developed non-peptide CGRP receptor antagonist BIBN4096BS for their potential to both inhibit the development and reverse tolerance to the antinociceptive action of morphine. Repeated administration of intrathecal morphine (15 microg), once daily, produced a progressive decline of antinociceptive effect and an increase in the ED(50) value in the tailflick and paw pressure tests. Co-administration of CGRP(8-37) (4 microg) or BIBN4096BS (0.05, 0.1 microg) with morphine (15 microg) prevented the decline of antinociceptive effect and increase in ED(50) value in the tailflick test. Intrathecal administration of the CGRP receptor antagonists did not alter the baseline responses in either tests. Acute CGRP(8-37) also did not potentiate the acute actions of spinal morphine. In animals rendered tolerant to intrathecal morphine, subsequent administration of CGRP(8-37) (4 microg) with morphine (15 microg) partially restored the antinociceptive effect and ED(50) value of acute morphine, reflecting the reversal of tolerance. Animals tolerant to intrathecal morphine expressed increased CGRP and substance P-like immunostaining in the dorsal horn of the spinal cord. The increase in CGRP, but not substance P-like immunostaining, was blocked by a co-treatment with CGRP(8-37) (4 microg). In animals already tolerant to morphine, the increase in CGRP but not substance P like immunostaining was partially reversed by CGRP(8-37) (4 microg). These data suggest that activation of spinal CGRP receptors contributes to both the development and expression of spinal opioid tolerance. CGRP receptor antagonists may represent a useful therapeutic approach for preventing as well as reversing opioid tolerance. PMID- 11053207 TI - Non-competitive pharmacological antagonism at the rabbit B(1) receptor. AB - The B(1) receptor for kinins, stimulated by kinin metabolites without the C terminal Arg residue (e.g., des-Arg(9)-bradykinin (BK) and Lys-des-Arg(9)-BK), is an increasingly recognized molecular target for the development of analgesic and anti-inflammatory drugs. Recently developed antagonists of this receptor were compared to a conventional antagonist, Ac-Lys-[Leu(8)]-des-Arg(9)-BK, in pharmacological assays based on the rabbit B(1) receptor. B-9858 (Lys-Lys [Hyp(3), Igl(5), D-Igl(7), Oic(8)]des-Arg(9)-BK) and three other analogues possessing the alpha-2-indanylglycine(5) (Igl(5)) residue (order of potency B 9858 approximately B-10146>B-10148>B-10050) were partially insurmountable antagonists of des-Arg(9)-BK in the contractility assay based on rabbit aortic rings. B-9858-induced depression of the maximal effect was more pronounced in tissues treated with the protein synthesis inhibitor cycloheximide to block the spontaneous increase of response attributed to the post-isolation formation of B(1) receptors, and only partly reversible on washing. By comparison, Ac-Lys [Leu(8)]des-Arg(9)-BK was a surmountable antagonist (pA(2) 7. 5), even in cycloheximide-treated tissues. B-9958 (Lys-[Hyp(3), CpG(5), D-Tic(7), CpG(8)]des Arg(9)-BK) was also surmountable (pA(2) 8.5). The binding of [(3)H]-Lys-des Arg(9)-BK to recombinant rabbit B(1) receptors expressed in COS-1 cells was influenced by two of the antagonists: while Ac-Lys-[Leu(8)]des-Arg(9)-BK competed for the radioligand binding without affecting the B(max), B-9858 decreased the B(max) in a time-dependent and washout-resistant manner. B-9858 and analogues possessing Igl(5) are the first reported non-competitive, non-equilibrium antagonists of the kinin B(1) receptor. PMID- 11053208 TI - Pharmacological examination of contractile responses of the guinea-pig isolated ileum produced by mu-opioid receptor antagonists in the presence of, and following exposure to, morphine. AB - We have assessed the potential of several mu-opioid receptor antagonists to elicit a response in the guinea-pig isolated ileum in the presence of, and following overnight exposure to, morphine. Naloxone, D-Phe-Cys-Tyr-D-Trp-Orn-Thr Pen-Thr-NH(2) (CTOP), (-)-5, 9alpha-diethyl-2-(3-furyl-methyl)-2'-hydroxy-6,7 benzomorphan (MR2266), but not D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH(2) (CTAP), produced a transient inhibition of electrically-evoked contractions of the guinea pig ileum. The effect of 1 microM CTOP, but not that to MR2266, was inhibited by 1 microM somatostatin. Naloxone (0.3 microM), CTOP (3 microM), CTAP (3 microM) and MR2266 (0.3 microM) antagonized the inhibitory effect of morphine on electrically-evoked contractions of the guinea-pig to a similar degree and, following 60 min exposure to morphine, produced non-sustained contractions. The response to 3 microM CTOP was significantly smaller than that to 3 microM CTAP. None of the antagonists produced a response in the absence of morphine. Following overnight exposure of the ileum to 0.3 microM morphine (4 degrees C), and repeated washing to remove the agonist, all four antagonists elicited non sustained contractions. However, the responses to 3 microM CTOP and 0.3 microM MR2266 were significantly smaller than those elicited by 0.3 microM naloxone and 3 microM CTAP. Somatostatin (1 microM) significantly reduced naloxone-induced contractions, but not those to CTAP. While all four mu-opioid antagonists elicited contractions in the presence of, and following prolonged exposure to, morphine, differences between them were noted which may be a consequence of non opioid actions. PMID- 11053209 TI - (8-Naphthalen-1-ylmethyl-4-oxo-1-phenyl-1,3,8-triaza-spiro[4. 5]dec-3-yl)-acetic acid methyl ester (NNC 63-0532) is a novel potent nociceptin receptor agonist. AB - Spiroxatrine was identified as a moderately potent (K:(i)=118 nM) but non selective agonist at the human nociceptin/orphanin FQ receptor, ORL1. This compound was subject to chemical modification and one of the resulting compounds, (8-naphthalen-1-ylmethyl-4-oxo-1-phenyl-1,3,8-triaza-s piro[4. 5]dec-3-yl)-acetic acid methyl ester (NNC 63-0532) was shown to have high affinity for ORL1 (K:(i)=7.3 nM). NNC 63-0532 showed only moderate affinity for the following receptors (K:(i) values in parentheses): mu-opioid (140 nM), kappa-opioid (405 nM), dopamine D(2S) (209 nM), dopamine D(3) (133 nM) and dopamine D(4.4) (107 nM) out of 75 different receptors, ion-channels and transporters. In functional assays, NNC 63-0532 was shown to be an agonist at ORL1 (EC(50)=305 nM), a much weaker agonist at the mu-opioid receptor (EC(50)>10 microM) and an antagonist or weak partial agonist at dopamine D(2S) (IC(50)=2830 nM). Thus, NNC 63-0532 is a novel non-peptide agonist with approximately 12 fold selectivity for ORL1 and may be useful for exploring the physiological roles of this receptor owing to its brain-penetrating properties. PMID- 11053210 TI - Release of a soluble ATPase from the rabbit isolated vas deferens during nerve stimulation. AB - The properties of the ATPase released during electrical field stimulation (EFS) (8 Hz, 25 s) of the sympathetic nerves of the superfused rabbit isolated vas deferens were investigated. Superfusate collected during EFS rapidly metabolised exogenous ATP (100 microM) and 50% was broken down in 5.67+/-0.65 min. The main metabolite was ADP, virtually no AMP was produced and adenosine was absent. No enzyme activity was seen in samples collected in the absence of EFS. Lineweaver Burke analysis of the initial rates of ATP hydrolysis gave a K(M) of 40 microM and V(max) of 20.3 nmol ATP metabolized min(-1) ml(-1) superfusate. ATPase activity was unaffected by storage at room temperature for 24 h, but was abolished at pH4 or by heating at 80 degrees C for 10 min. ARL 67156 inhibited ATP breakdown in a concentration-dependent manner (IC(50)=25 microM (95% confidence limits=22-27 microM), Hill slope=-1.06+/-0.04). When EFS was applied three times at 30 min intervals, ATP metabolism was 20-30% less in superfusate collected during the second and third stimulation periods compared with the first. ATPase activity was released in a frequency-dependent manner, with significantly greater activity seen after stimulation at 4 and 8 Hz than at 2 Hz. In conclusion, EFS of the sympathetic nerves in the rabbit vas deferens causes release of substantial ATPase, but little ADPase activity into the extracellular space. This contrasts with the guinea-pig vas deferens, which releases enzymes that degrade ATP to adenosine. Thus, the complement of enzymes released by nerve stimulation is species-dependent. PMID- 11053211 TI - Beta-agonist-induced constitutive beta(2)-adrenergic receptor activity in bovine tracheal smooth muscle. AB - According to the two state receptor model, the beta(2)-adrenergic receptor (beta(2)-AR) isomerizes between an inactive state and a constitutively active state, which couples to the stimulatory G-protein in the absence of agonist. In bovine tracheal smooth muscle (BTSM), we investigated the effect of short and long term beta(2)-AR activation by fenoterol on constitutive receptor activity. Preincubation of the BTSM strips for 5 min, 30 min and 18 h with 10 microM fenoterol, followed by extensive washout (3 h, 37 degrees C), caused a rapid and time-dependent inhibition of KCl-induced contraction, reaching 68+/-10, 51+/-6 and 46+/-4% of control, respectively, at 40 mM KCl (P:<0.05 all). At all time points, the EC(50) values to KCl were significantly reduced as well. Preincubation of BTSM with 0.1, 1.0 and 10 microM fenoterol during 18 h caused a concentration-dependent decrease of the 40 mM KCl response to 70+/-5, 47+/-12 and 43+/-9% of control, respectively (P:<0.05 all). The reduced KCl contractions were reversed in the presence of 1 microM timolol. Moreover, the sensitivity to KCl in the presence of timolol was enhanced after fenoterol incubation. Inverse agonism was also found for other beta-blockers, with a rank order of efficacy of pindolol >/=timolol=propranolol>alprenolol>/=sotalol>labetalol. At 25 mM KCl-induced tone, the contraction induced by cumulative timolol administration was competitively antagonized by the less efficacious inverse agonist labetalol, indicating that the fenoterol-induced effects cannot be explained by residual beta-agonist binding. In conclusion, fenoterol treatment of BTSM causes a time- and concentration-dependent development of constitutive beta(2)-AR activity, which can be reversed by various inverse agonists. The beta-agonist-induced changes could represent a novel regulation mechanism of beta(2)-AR activity. PMID- 11053212 TI - Antidystonic efficacy of nitric oxide synthase inhibitors in a rodent model of primary paroxysmal dystonia. AB - In a hamster model (genetic symbol dt(sz)) of primary paroxysmal non-kinesiogenic dystonic choreoathetosis, recent studies have shown beneficial effects of glutamate and dopamine receptor antagonists. Nitric oxide (NO), synthesized from L-arginine by NO synthase in response to glutamate receptor activation, elicits cyclic GMP and modulates glutamate-mediated processes and striatal dopamine release. Therefore, the effects of NO synthase inhibitors and of L-arginine on severity of dystonia were investigated in dt(sz) hamsters in which dystonic attacks, characterized by twisting movements and postures, can be induced by stress. The NO synthase inhibitors N(G)-nitro-L-arginine (L-NNA), N(G)-nitro-L arginine methyl ester (L-NAME) and 7-nitroindazole significantly reduced the severity of dystonia. At antidystonic effective doses neither L-NNA nor L-NAME caused observable side effects, whereas 7-nitroindazole exerted moderate reduction of locomotor activity. The antidystonic effect of L-NAME was reversed by co-administration of the NO precursor L-arginine. However, L-arginine administered alone did not exert any effect on severity of dystonia. Cerebellar cyclic GMP levels in brains of mutant hamsters in comparison to non-dystonic control hamsters did not significantly differ, but the cerebellar cyclic GMP levels tended to be increased in dt(sz) hamsters during a dystonic attack. L-NAME significantly decreased the cerebellar cyclic GMP levels in both dt(sz) and control hamsters. Although an overproduction of NO is probably not critically involved in the pathogenesis of paroxysmal dystonia, it may contribute to the manifestation of dystonic attacks, as indicated by the antidystonic effects of NO synthase inhibitors. Peripheral side effects may limit the clinical use of NO synthase inhibitors, but more selective inhibitors of the neuronal NO synthase should be considered as interesting candidates for the treatment of paroxysmal dystonia. PMID- 11053213 TI - 5-HT(4) receptors on cholinergic nerves involved in contractility of canine and human large intestine longitudinal muscle. AB - 5-HT(4) receptors mediate circular muscle relaxation in both human and canine large intestine, but this phenomenon alone can not explain the improvement in colonic motility induced by selective 5-HT(4) receptor agonists in vivo. We set out to characterize 5-HT(4) receptor-mediated effects in longitudinal muscle strips of canine and human large intestine. Electrical field stimulation (EFS) was applied providing submaximal isotonic contractions. L-NOARG (0.1 mM) was continuously present in the organ bath to preclude nitric oxide-induced relaxation to EFS. The selective 5-HT(4) receptor agonist prucalopride (0.3 microM) enhanced EFS-evoked contractions, that were antagonized in both preparations by the selective 5-HT(4) receptor antagonist GR 113808 (0.1 microM). The prucalopride-induced increase was present in canine ascending and descending colon, but absent in rectum. Regional differences in response to prucalopride were not observed in human ascending and sigmoid colon and rectum. Incubation with atropine (1 microM) or tetrodotoxin (0.3 microM) inhibited EFS-induced contractions, which were then unaffected by prucalopride (0.3 microM) in both tissues. In the presence of methysergide (3 microM; both tissues) and granisetron (0.3 microM; only human tissues), 5-HT (0.3 microM) enhanced EFS-induced contractions, an effect that was antagonized by GR 113808 (0.1 microM). In the presence of atropine or tetrodotoxin, EFS-induced contractions were inhibited, leaving 5-HT (0.3 microM) ineffective in both preparations. This study demonstrates for the first time that in human and canine large intestine, 5-HT(4) receptors are located on cholinergic neurones, presumably mediating facilitating release of acetylcholine, resulting in enhanced longitudinal muscle contractility. This study and previous circular muscle strip studies suggest that 5-HT(4) receptor agonism facilitates colonic propulsion via a coordinated combination of inhibition of circumferential resistance and enhancement of longitudinal muscle contractility. PMID- 11053214 TI - A comparison of agonist-specific coupling of cloned human alpha(2)-adrenoceptor subtypes. AB - The agonist-specific coupling properties of the three cloned human alpha(2) adrenoceptor subtypes have been compared, when expressed at similar levels in Chinese hamster ovary (CHO) cell lines, using noradrenaline and (+/-)-meta octopamine as agonists. Noradrenaline can couple the receptor to both the inhibition and stimulation of forskolin-stimulated cyclic AMP production in all three receptor subtypes, with the relative strength of the coupling to the pathways varying for each of the receptor subtypes. meta-Octopamine selectively couples the alpha(2A)-adrenoceptor only to the inhibition of forskolin-stimulated cyclic AMP production. However, meta-octopamine couples the alpha(2B)- and alpha(2C)-adrenoceptors to both the inhibition and stimulation of forskolin stimulated cyclic AMP production. The relative potency of meta-octopamine to noradrenaline varies between the different alpha(2)-adrenoceptor subtypes. The effects of meta-octopamine are around two orders of magnitude less potent than those of noradrenaline on both the alpha(2A)- and alpha(2B)-adrenoceptor subtypes. In contrast, in the case of the alpha(2C)-adrenoceptor, meta-octopamine is only one order of magnitude less potent than noradrenaline in the stimulation of forskolin-stimulated cyclic AMP production and, in addition, is equipotent with noradrenaline in the inhibition of forskolin-stimulated cyclic AMP production and has an increased maximal response. This raises the possibility that meta-octopamine may have physiologically important actions via alpha(2C) adrenoceptors in vivo. The results show that the modulation of cyclic AMP production occurs in both a subtype- and agonist-specific manner for alpha(2A) adrenoceptors and in a subtype specific manner for alpha(2B)- and alpha(2C) adrenoceptors. PMID- 11053215 TI - Increase in participation of vasoactive intestinal peptide in relaxation of the distal colon of Wistar rats with age. AB - Changes in participation of vasoactive intestinal peptide (VIP) in nonadrenergic noncholinergic (NANC) relaxation of longitudinal muscle of the distal colon with age were studied in 2- to 50-week-old Wistar rats in vitro. The extent of the VIP mediated component of the relaxation induced by electrical field stimulation (EFS) was determined by the effect of VIP(10 - 28), a VIP receptor antagonist. In 2-week-old rats, the extent of the VIP-mediated component of the relaxation was scarce, about 10%, whereas the component gradually increase with age and reached the maximum extent 66% at 50-week-old. Since our previous results suggest that VIP induces NANC relaxation via activation of charybdotoxin (ChTx, a blocker of large conductance Ca(2+)-activated K(+) channel)-sensitive K(+) channels with concomitant slow hyperpolarization in the muscle cells, we next studied whether ChTx-sensitive component and slow hyperpolarization changes with age. Extent of ChTx-sensitive component of the relaxation increased with age, showing a very similar pattern to VIP-mediated one. EFS induced monophasic inhibitory junction potentials (i.j.ps) in longitudinal muscle cells of the distal colon of 2- and 4 week-old. EFS also induced biphasic i.j.ps in many longitudinal muscle cells of 8 and 50-week-old: rapid and subsequent slow hyperpolarization. A VIP receptor antagonist selectively inhibited the slow hyperpolarization. Exogenously added VIP induced no appreciable change in the membrane potential of longitudinal muscle cells of 2-week-old, whereas it induced slight slow hyperpolarization of the cell membrane in 4-week-old and magnitude of the hyperpolarization increased with age. On the other hand, relaxant response of the longitudinal muscle to exogenously added VIP was high in younger rats. The present results suggest that the role of VIP in mediating NANC relaxation of longitudinal muscle of the Wistar rat distal colon is very little at neonatal stage, but it increases with age. PMID- 11053216 TI - Molecular cloning, sequence analysis and pharmacological properties of the porcine 5-HT(1D) receptor. AB - A cDNA encoding the full-length 5-HT(1D) receptor derived from porcine cerebral cortex was amplified, cloned and sequenced, using guinea-pig 5-HT(1D) receptor coding sequence oligonucleotide primers in reverse transcription-polymerase chain reaction (RT - PCR). The 5' and 3' ends of the porcine 5-HT(1D) receptor cDNA were verified by inverse PCR. Sequence analysis of porcine 5-HT(1D) receptor cDNA revealed an open reading frame of 1134 nucleotides encoding a polypeptide of 377 amino acids having 92% homology with the human 5-HT(1D) receptor and 88 - 90% homology with other species homologues. The porcine 5-HT(1D) receptor cDNA was further subcloned into a mammalian expression vector pcDNA3 and expressed in monkey Cos-7 cells. Radioligand binding assays using either [(3)H]-5-CT or [(3)H] GR125743 on Cos-7 cell membranes showed that pK(i) values of 14 serotonin ligands were highly correlated with those obtained with the human 5-HT(1D) receptor. Nonetheless, a selective antagonist at the human 5-HT(1D) receptor, BRL15572, only poorly recognized the porcine homologue. Using membranes from cells co expressing the porcine 5-HT(1D) receptor and rat G(alphail)Cys(351) Ile protein, it was shown that 5-HT and zolmitriptan increased, while ketanserin decreased basal [(35)S]-GTPgammaS binding. The potency of zolmitriptan in the [(35)S] GTPgammaS binding assay (pEC(50): 8. 46+/-0.08) agreed with its affinity in displacing the radioligands [(3)H]-5-CT and [(3)H]-GR125743 (pK(i): 8.38+/-0.15 and 8.67+/-0.08, respectively). In conclusion, we have established the cDNA sequence and pharmacology of the cloned porcine 5-HT(1D) receptor. This information would be useful in exploring the role of divergent amino acid residues in the receptor-ligand interaction as well as the role of 5-HT(1D) receptor in pathophysiological processes relevant for novel drug discovery in diseases such as migraine. PMID- 11053217 TI - Tetrahydrobiopterin impairs the action of endothelial nitric oxide via superoxide derived from platelets. AB - The mechanism by which exogenous tetrahydrobiopterin (BH(4)) impairs the action of endothelial nitric oxide (NO) in the presence of platelets was investigated. The endothelial NO generated by shear stress was determined by the anti aggregating activity of indomethacin-treated endothelial cells and the cyclic GMP concentration in platelets. The inhibitory effect of exogenous BH(4) was suppressed by superoxide dismutase (SOD), or diclofenac sodium at concentrations inhibiting O(2)(-) generation, but not by allopurinol, a xanthine oxidase inhibitor. BH(4) similarly inhibited the anti-aggregatory effect of sodium nitroprusside (SNP), a NO donor. The inhibitory effect was suppressed by diphenyleneiodonium, a specific inhibitor of NADPH oxidase. Six(S)-BH(4), an inactive diastereoisomer of 6(R)-BH(4), and the 5,6,7,8-tetrahydropterin compounds inhibited the endothelial NO action, whereas sepiapterin and 7,8 dihydrobiopterin (BH(2)), 5,6-double bond pterins, were inactive. These tetrahydropterins, but not sepiapterin and BH(2), scavenged superoxide (O(2)(-)) generated by the hypoxanthine-xanthine oxidase reaction, possibly due to electron transfer during oxidation to its quinonoid-form. BH(4) markedly stimulated the O(2)(-) generation from platelets, in the presence of NADH, rather than that of NADPH. These findings suggest that BH(4) stimulates platelet NAD(P)H oxidase to generate O(2)(-), and inhibits the anti-aggregating effect of NO. SOD activity in the local environment may modify the effect of BH(4) on the endothelial NO activity. PMID- 11053218 TI - An evaluation of potassium ions as endothelium-derived hyperpolarizing factor in porcine coronary arteries. AB - In the rat hepatic artery, the endothelium-derived hyperpolarizing factor (EDHF) was identified as potassium. Potassium hyperpolarizes the smooth muscles by gating inward rectified potassium channels and by activating the sodium-potassium adenosine triphosphatase (Na(+)-K(+)ATPase). Our goal was to examine whether potassium could explain the EDHF in porcine coronary arteries. On coronary strips, the inhibition of calcium-dependent potassium channels with 100 nM apamin plus 100 microM charibdotoxin inhibited the endothelium-dependent relaxations, produced by 10 nM substance P and 300 nM bradykinin and resistant to nitro-L arginine and indomethacin. The scavenging of potassium with 2 mM Kryptofix 2.2.2 abolished the endothelium-dependent relaxations produced by the kinins and resistant to nitro-L-arginine and indomethacin. Forty microM 18alpha glycyrrethinic acid or 50 microM palmitoleic acid, both uncoupling agents, did not inhibit these kinin relaxations. Therefore, EDHF does not result from an electrotonic spreading of an endothelial hyperpolarization. Barium (0.3 nM) did not inhibit the kinin relaxations resistant to nitro-L-arginine and indomethacin. Therefore, EDHF does not result from the activation of inward rectified potassium channels. Five hundred nM ouabain abolished the endothelium-dependent relaxations resistant to nitro-L-arginine and indomethacin without inhibiting the endothelium derived NO relaxation. The perifusion of a medium supplemented with potassium depolarized and contracted a coronary strip; however, the short application of potassium hyperpolarized the smooth muscles. These results are compatible with the concept that, in porcine coronary artery, the EDHF is potassium released by the endothelial cells and that this ion hyperpolarizes and relaxes the smooth muscles by activating the Na(+)-K(+)ATPase. PMID- 11053219 TI - Effects of a selective 5-HT(1B/1D) receptor agonist on spinal and trigeminal reflexes in the anaesthetized rabbit. AB - The effects of the 5-HT(1B/1D) receptor agonist L-741,604 on a trigeminally mediated (jaw depressor) reflex and a spinally-mediated (flexion withdrawal) reflex have been compared between spinalized and intact, anaesthetized rabbits. L 741,604 depressed the jaw depressor reflex dose-dependently in all animals, to a median of 5% (inter-quartile range, IQR, 3 - 28%, n=18) of pre-drug levels after a cumulative dose of 3.1 micromol kg(-1) i.v. This effect was reversed by the 5 HT(1B/1D) antagonist GR 127,935 (1 - 2 micromol kg(-1) i.v.). The flexion withdrawal reflex was depressed by L-741, 604 in non-spinalized animals, to a median of 22% (IQR 10 - 36%, n=10) of pre-drug levels after the highest dose, an action that was reversed by GR 127,935. In spinalized rabbits, L-741,604 up to 0.3 micromol kg(-1) i.v. cumulative increased the flexion reflex to a median of 189% (IQR 169 - 198%, n=8) of pre-drug controls. With higher doses the reflex decreased, so that after 3.1 micromol kg(-1) it was 75% (IQR 55 - 96%) of pre drug levels. Subsequent GR 127,935 increased reflexes to a median of 180% (IQR 136 - 219%) of controls. L-741,604 increased arterial blood pressure and decreased heart rate in both preparations, effects that were reversed by GR 127,935. Thus, when the spinal cord was intact L-741,604 inhibited spinal and trigeminal reflexes in the same way. Although spinalization enabled a non-5 HT(1B/1D)-mediated excitatory effect of L-741,604 on spinal reflexes, there was a clear inhibitory effect of the drug at high doses. These data suggest that L 741,604 inhibits spinal reflexes by increasing descending inhibition and by a direct action in the cord. The same processes could apply to inhibition of trigeminally-mediated events. PMID- 11053220 TI - Extracellular signal-regulated kinase 1/2 control Ca(2+)-independent force development in histamine-stimulated bovine tracheal smooth muscle. AB - The role of extracellular signal-regulated kinase (ERK)-1 and ERK-2 in controlling histamine-induced tone in bovine trachealis was investigated. PD 098059, an inhibitor of mitogen-activated protein kinase kinase (MKK)-1, had no effect on the histamine concentration-response relationship that described contraction. However, in the presence of EGTA, PD 098059 produced a parallel 5 fold rightwards shift of the histamine concentration-response curve without reducing the maximum response. The beta(2)-adrenoceptor agonist, procaterol, also displaced the histamine-concentration response curve to the right but the effect was much greater than that evoked by PD 098059, non-competitive and seen in the absence and presence of EGTA. A low basal level of pERK-1 and pERK-2 was always detected in untreated trachealis, which was significantly higher in EGTA-treated tissues and inhibited by PD 098059 and procaterol. Histamine markedly enhanced the phosphorylation of ERK-1 and ERK-2 by a mechanism that was also enhanced by EGTA and significantly attenuated by procaterol and PD 098059. Neither cholera toxin nor SP:-8-Br-cAMPS mimicked the ability of procaterol to dephosphorylate ERK. Similarly, neither pertussis toxin (PTX) nor RP:-8-Br-cAMPS, an inhibitor of cyclic AMP-dependent protein kinase (PKA), affected basal pERK levels or antagonized the inhibitory effect of procaterol. These data implicate the MKK 1/ERK signalling cascade in Ca(2+)-independent, histamine-induced contraction of bovine trachealis. In addition, the ability of procaterol to dephosphorylate ERK in an RP:-8-Br-cAMPS- and PTX-insensitive manner suggests that this may contribute to the anti-spasmogenic activity of beta(2)-adrenoceptor agonists by activating a novel PKA-independent pathway. PMID- 11053221 TI - Chronic, but not acute, dosing of antipsychotic drugs alters neurotensin binding in rat brain regions. AB - The present study compared high affinity neurotensin (NT) binding in rat brain following acute or chronic treatment with the classical antipsychotic, haloperidol, and the newer antipsychotic drugs, clozapine and zotepine. Drugs were given orally, as an acute treatment (1 dose) or chronically (21 day dosing) and binding to the NT high affinity receptor was examined in three brain regions; striatum, nucleus accumbens/olfactory tubercle and frontal cortex. Acute dosing with either vehicle, haloperidol, clozapine or zotepine produced no significant changes in NT binding from controls (naive rats). Chronic (21 day) dosing resulted in an increase in the K:(D:) and B(max) of high affinity receptors in the striatum following haloperidol, but not clozapine, zotepine or vehicles. In contrast, the newer antipsychotics, clozapine and zotepine but not haloperidol or vehicles, significantly altered NT binding in the nucleus accumbens/olfactory tubercle by decreasing the K:(D:) and B(max). Further differentiation between the two newer antipsychotic drugs occurred in the frontal cortex. Clozapine had no significant effect on NT binding, whereas zotepine significantly reduced the K:(D:) of the high affinity receptor with no alteration in B(max). The antipsychotic drugs tested did not interact directly with the NT high affinity receptor. Therefore, they must be acting indirectly via an alternative receptor mechanism to alter NT high affinity binding. In accordance with previously reported NT/dopamine receptor interactions, this would suggest cross-talk between these systems. Overall, these data demonstrate that chronic, but not acute, administration of antipsychotic drugs alters NT binding in the rat brain. In addition, anatomical differences in NT binding arise according to the antipsychotic drug under test. This may be predictive of drug side-effect profile, antipsychotic efficacy or atypicality. PMID- 11053222 TI - Mecamylamine but not the alpha7 receptor antagonist alpha-bungarotoxin blocks sensitization to the locomotor stimulant effects of nicotine. AB - The involvement of alpha7 receptors in the locomotor stimulant effects of nicotine has been examined by determining the ability of intracerebroventricular (i.c.v.) administration of the alpha7 receptor antagonist alpha-bungarotoxin (alpha-bgt) to modify sensitization to the locomotor activating effects of chronic nicotine. Intracerebroventricular administration of alpha-bgt (0.02 - 8 nmoles) produced a dose dependent increase in convulsive behaviour. At doses less than 1.0 nmole, minimal convulsive behaviour occurred but larger doses evoked convulsions in all rats which displayed a more rapid onset time as the dose increased. The binding distribution of alpha7 receptors 20 min and 3 h following an i.c.v. administration of [(125)I]-alpha-bgt (0.02 nmoles) revealed clear binding in the hippocampus, cingulate cortex and hypothalamus which was more intense after 3 h. Rats chronically treated with nicotine (0.4 mg kg(-1)) and exposed to the locomotor activity apparatus daily acquired an increase in locomotor activity relative to the control group after 3 days of treatment which reached a maximum after 7 days of treatment and was maintained for the 2 week treatment period. Pre-treatment with mecamylamine (1 mg kg(-1)) prevented the expression of the locomotor stimulant effects of nicotine but pre-treatment with i.c.v. alpha-bgt (0.02 nmoles) did not affect nicotine-induced changes in locomotor activity. The results of this study support the conclusion that nicotinic receptors of the alpha4beta2 subtype rather than the alpha7 subtype are important in mediating the expression of the locomotor stimulant effects of nicotine. PMID- 11053223 TI - Transforming growth factor-alpha stimulates prostaglandin generation through cytosolic phospholipase A(2) under the control of p11 in rat gastric epithelial cells. AB - The regulatory effects of transforming growth factor (TGF)-alpha on phospholipase A(2) (PLA(2)) isozymes contributing to prostaglandin generation in rat gastric epithelial RGM1 cells were examined. Stimulation with TGF-alpha for 24 h time dependently induced prostaglandin E(2) generation with an increase in cyclo oxygenase-2 protein. The TGF-alpha-induced prostaglandin E(2) generation was suppressed by NS-398, a cyclo-oxygenase-2 inhibitor. TGF-alpha stimulated the activity and the protein synthesis of cytosolic PLA(2) (cPLA(2)). A time dependent increase in cPLA(2) protein occurred in parallel with PGE(2) generation, which was inhibited by methyl arachidonyl fluorophosphonate (MAFP), a cPLA(2) inhibitor. However, no change in activity of secretory PLA(2) or Ca(+2) independent PLA(2) was observed in the TGF-alpha-stimulated cells. Stimulation with the Ca(2+) ionophore A23187 for 10 min induced MAFP-sensitive arachidonic acid liberation. Interestingly, preincubation with TGF-alpha for 24 h diminished A23187-stimulated arachidonic acid liberation despite the increase in cPLA(2) protein. Under the conditions, TGF-alpha was found to increase p11, an endogenous cPLA(2) suppressor, also known as annexin II light chain. The TGF-alpha-induced increase in p11 was suppressed by tyrphostin AG1478, an inhibitor of tyrosine kinase of epidermal growth factor receptor, which was also found to restore the inhibition by TGF-alpha of A23187-stimulated arachidonic acid liberation. However, TGF-alpha did not alter protein levels of annexin II heavy chain. These results suggest that TGF-alpha stimulates prostaglandin generation through an increase in cPLA(2), the hydrolytic action of which may be under the control of p11. PMID- 11053224 TI - Activation of phospholipase D by metabotropic glutamate receptor agonists in rat cerebrocortical synaptosomes. AB - The pharmacological profile of metabotropic glutamate receptor (mGluR) activation of phospholipase D (PLD), and the associated signalling pathways, were examined in rat cerebrocortical synaptosomes. The assay was conducted using a transphosphatidylation reaction in synaptosomes which were pre-labelled with either [(3)H]-arachidonic acid or [(32)P]-orthophosphate. The mGluR agonists (1S,3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S, 3R-ACPD) and (RS)-3,5 dihydroxyphenylglycine (DHPG), both activated PLD, while phorbol 12,13-dibutyrate (PDBu) treatment caused receptor-independent activation of PLD and had an additive effect on 1S,3R-ACPD induced PLD activity. A protein kinase C (PKC) inhibitor, GF109203X, failed to antagonize mGluR receptor-coupled PLD activity. We could not detect any increase in the products of PI (phosphoinositide) specific phospholipase C (PI-PLC), inositol(1,4, 5)trisphosphate or diacylglycerol, by 1S, 3R-ACPD at 15 s. However, diacylglycerol increased monophasically in response to mGluR agonists and remained elevated for at least 15 min. Phosphatidic acid phosphohydrolase (PAP) activity, which converts PA to DAG, was present in the synaptosomes. These data suggest that, in rat cerebrocortical synaptosomes, the 1S,3R-ACPD-sensitive mGluR is coupled to PLD through a mechanism that is independent of both PKC and PI-PLC. PMID- 11053226 TI - Corrigendum PMID- 11053225 TI - Endothelial nitric oxide synthase is a site of superoxide synthesis in endothelial cells treated with glyceryl trinitrate. AB - Tolerance to glyceryl trinitrate (GTN) involves superoxide (O(2)(*-)) production by endothelial cells. Nitric oxide synthase (NOS) produces O(2)(*-) when L arginine (L-arg) is limited. The purpose of this study was to test the hypothesis that GTN stimulates NOS to increase O(2)(*-) synthesis in endothelial cells when L-arg is limited. Production of O(2)(*-) by bovine aortic endothelial cells (BAEC, passages 3 - 5) was determined by spectrophotometrically measuring superoxide dismutase-inhibited reduction of ferricytochrome C to ferrocytochrome C. Cells were incubated in buffer without L-arg. O(2)(*-) production was measured using BAEC either untreated or treated with L-NAME or L-arg alone or following treatment with GTN (10(-9) to 10(-6) M) for 30 min or DPTA NONOate (10(-7) and 10(-6) M) alone or with GTN or DPTA NONOate after pretreatment with nitro-L arginine methyl ester (L-NAME), L-arg or their inactive enantiomers, D-NAME or D arg (all 5 x 10(-4) M) (n=6 - 7/group). L-NAME alone produced a 69% reduction in O(2)(*-) levels. Treatment with L-arg alone had no effect. Cells treated with GTN alone exhibited an increase in O(2)(*-). This effect was prevented by pretreatment with either L-NAME or L-arg, and was unaffected by D-NAME or D-arg. We observed a dose-response relationship in O(2)(*-) production to GTN over a range of 10(-9) to 10(-7) M. The NO donor, DPTA-NONOate, unlike GTN, did not have a significant effect on O(2)(*-) production. In conclusion, endothelial NOS is a site of O(2)(*-) synthesis in endothelial cells activated by GTN. PMID- 11053227 TI - The multiple roles of visual cortical areas MT/MST in remembering the direction of visual motion. AB - Although the role of cortical areas MT and MST (MT/MST) in the processing of directional motion information is well established, little is known about the way these areas contribute to the execution of complex behavioral tasks requiring the use of such information. We tested monkeys with unilateral lesions of these areas on a visual working memory task in which motion signals not only had to be encoded, but also stored for brief periods of time and then retrieved. The monkeys compared the directions of motion of two random-dot stimuli, sample and test, separated by a temporal delay. By increasing the temporal delay and spatially separating the two stimuli, placing one in the affected visual field and the other in the intact visual field, we were able to assess the contribution of MT/MST to specific components of the task: encoding (sample), retention (delay) and encoding/retrieval/comparison (test). We found that the effects of MT/MST lesions on specific components depended upon the demands of the task and the nature of the visual motion stimuli. Whenever stimuli consisted of random dots moving in a broad range of directions, MT/MST lesions appeared to affect encoding. Furthermore, when the lesions affected encoding of the sample, retention of the direction of stimulus motion was also affected. However, when the stimulus was coherent and the emphasis of the task was on the comparison of small direction differences, the absence of MT/MST had major impact on the retrieval/comparison component of the task and not on encoding or storage. PMID- 11053228 TI - Translaminar differentiation of visually guided behaviors revealed by restricted cerebral cooling deactivation. AB - The purpose of the present study was to test the hypothesis that superficial and deep layers within a single cerebral region influence cerebral functions and behaviors in different ways. For this test, we selected posterior middle suprasylvian (pMS) sulcal cortex of the cat, a suspected homolog of the area V5 complex of primates, because the region has been implicated in several visually guided behaviors. Cats were trained on three tasks: (1) discrimination of direction of motion; (2) discrimination of static patterns partially obscured by static or moving masks; and (3) visual detection and orienting. Cooling of cryoloops in contact with pMS sulcal cortex to 8+/-1 degrees C selectively and completely impaired performance on the two motion discrimination tasks (1 and 2), while leaving the detection and orienting task (task 3) unimpaired. Further cooling to 3 degrees C resulted in an additional complete impairment of task 3. The 8 degrees C temperature resulted in silencing of neuronal activity in the supragranular layers (I-III) and the 3 degrees C temperature silenced activity throughout the thickness of pMS sulcal cortex. The variation in behavioral performance with covariation of cryoloop temperature and vertical, but not lateral, spread of deactivation shows that deactivation of superficial cerebral layers alone was sufficient to completely impair performance on the two motion discrimination tasks, whereas additional deactivation of the deep layers was essential to block performance on the detection and orienting task. Thus, these results show a functional bipartite division of labor between upper and lower cortical layers that is supported by efferent connectional anatomy. Similar bipartite division into upper and lower layers may be a general feature of cerebral cortical architecture, signal processing and guidance of behavior. PMID- 11053229 TI - Physiological dysfunction of the dorsolateral prefrontal cortex in schizophrenia revisited. AB - Evidence implicates subtle neuronal pathology of the prefrontal cortex (PFC) in schizophrenia, but how this pathology is reflected in physiological neuroimaging experiments remains controversial. We investigated PFC function in schizophrenia using functional magnetic resonance imaging (fMRI) and a parametric version of the n-back working memory (WM) task. In a group of patients who performed relatively well on this task, there were three fundamental deviations from the 'healthy' pattern of PFC fMRI activation to varying WM difficulty. The first characteristic was a greater magnitude of PFC fMRI activation in the context of slightly impaired WM performance (i.e. physiological inefficiency). The second was that the significant correlations between behavioral WM performance and dorsal PFC fMRI activation were in opposite directions in the two groups. Third, the magnitude of the abnormal dorsal PFC fMRI response was predicted by an assay of N-acetylaspartate concentrations (NAA) in dorsal PFC, a measure of neuronal pathology obtained using proton magnetic resonance spectroscopy. Patients had significantly lower dorsal PFC NAA than controls and dorsal PFC NAA inversely predicted the fMRI response in dorsal PFC (areas 9, 46) to varying WM difficulty supporting the assumption that abnormal PFC responses arose from abnormal PFC neurons. These data suggest that under certain conditions the physiological ramifications of dorsal PFC neuronal pathology in schizophrenia includes exaggerated and inefficient cortical activity, especially of dorsal PFC. PMID- 11053230 TI - Partially overlapping neural networks for real and imagined hand movements. AB - Neuroimagery findings have shown similar cerebral networks associated with imagination and execution of a movement. On the other hand, neuropsychological studies of parietal-lesioned patients suggest that these networks may be at least partly distinct. In the present study, normal subjects were asked to either imagine or execute auditory-cued hand movements. Compared with rest, imagination and execution showed overlapping networks, including bilateral premotor and parietal areas, basal ganglia and cerebellum. However, direct comparison between the two experimental conditions showed that specific cortico-subcortical areas were more engaged in mental simulation, including bilateral premotor, prefrontal, supplementary motor and left posterior parietal areas, and the caudate nuclei. These results suggest that a specific neuronal substrate is involved in the processing of hand motor representations. PMID- 11053231 TI - Dynamics of striate cortical activity in the alert macaque: I. Incidence and stimulus-dependence of gamma-band neuronal oscillations. AB - Using single and multiunit recordings in the striate cortex of alert macaque monkeys, we find that gamma-band (20-70 Hz) oscillations in neuronal firing are a prominent feature of V1 neuronal activity. The properties of this rhythmic activity are very similar to those previously observed in the cat. Gamma-band activity is strongly dependent on visual stimulation, largely absent during spontaneous activity and, under the conditions of our experiment, not time-locked to the vertical refresh of the computer monitor (80 Hz) used to present the stimuli. In our sample, 61% of multiunit activity (MUA) and 46% of single-unit activity (SUA) was significantly oscillatory, with mean frequencies of 48+/-9 and 42+/-13 Hz, respectively. Gamma-band activity was most likely to occur when cells were activated by their optimal stimuli, but still occurred, although less often and with lower amplitude, in response to nonoptimal stimuli. The frequency of gamma-band activity also reflected stimulus properties, with drifting gratings evoking higher-frequency oscillations than stationary gratings. As in the cat, the spike trains of single cells showing gamma-band oscillations often displayed a pattern of repetitive burst firing, with intraburst firing rates of 300-800 Hz. The overall similarity of rhythmic neuronal activity in the primary visual cortex of cats and monkeys suggests that the phenomenon is not species-specific. The stimulus-dependence of the rhythmic activity is consistent with a functional role in visual perception. PMID- 11053232 TI - Dynamics of striate cortical activity in the alert macaque: II. Fast time scale synchronization. AB - Synchronous neuronal activity with millisecond precision has been postulated to contribute to the process of visual perceptual grouping. We have performed multineuron recordings in striate cortex of two alert macaque monkeys to determine if the occurrence and properties of this form of activity are consistent with the minimal requirements of this theory. We find that neuronal synchronization with millisecond precision is a prevalent and robust feature of stimulus-evoked activity in striate cortex. It occurs among adjacent cells recorded by the same electrode (<120 microm), among cells recorded at separate but nearby sites (300-400 microm) and between cells recorded at locations separated by 3-4 mm. The magnitude and probability of synchronous firing is inversely related to the spatial separation between the cells and it occurs within and between groups of cells that are both tuned and untuned for stimulus orientation and direction. Among those tuned for orientation, cell pairs separated by <400 microm showed no clear dependence of correlated firing on orientation preference. The occurrence of gamma-band (20-70 Hz) oscillations in the cellular firing patterns was a strong predictor of synchronous firing at each of the spatial scales. Nearly 90% of the cell pairs showing significant correlation also showed oscillatory firing in one or both cells of the pair. These results are consistent with some, but not all, of the previous reports of synchronous activity in striate cortex of both cat and primates. The similarities in the properties of synchronous oscillations in the monkey and cat suggest that this form of neuronal activity is a general property of mammalian striate cortex. The relation between correlation and oscillation suggests that neuronal rhythmicity is an important mechanism contributing to synchronization. PMID- 11053233 TI - Glutamate, GABA and precursor amino acids in adult mouse neocortex: cellular diversity revealed by quantitative immunocytochemistry. AB - Glutamate is an important amino acid in the neocortex for metabolic and neurotransmitter functions. The objective of this study was to detect variations in cellular glutamate content using quantitative immunocytochemistry. We show that glutamate is present in almost all cortical cells and coexists with other amino acids such as aspartate, glutamine or gamma-aminobutyric acid (GABA). The patterns of aspartate and glutamine content suggests that there are no purely aspartatergic or glutaminergic neurons. GABAergic neurons showed variable levels of the precursors such as glutamate, glutamine and aspartate. Comparison of immunoreactive patterns between two cortical areas did not detect any statistically significant differences. The mean cellular intensity for GABA and glutamate was constant across different layers. Surprisingly, we found that GABAergic neurons could coexist with either low or high levels of glutamate, suggesting that metabolic levels of glutamate in these neurons could be variable. Alternatively, some GABA neurons may utilize both GABA and glutamate for neurotransmission. We show that when variations in amino acid content are separately mapped onto individual cells, co-registration is a useful technique for reporting heterogeneity among cortical cells. PMID- 11053234 TI - Differential short-term synaptic plasticity and transmission of complex spike trains: to depress or to facilitate? AB - Experimental studies have revealed conspicuous short-term facilitation and depression that are expressed differentially at distinct classes of cortical synapses. To explore computational implications of synaptic dynamics, we investigated transmission of complex spike trains through a stochastic model of cortical synapse endowed with short-term facilitation and vesicle depletion. Inputs to the synapse model were either real spike train data recorded from the visual and prefrontal cortices of behaving monkeys, or were generated numerically with prescribed temporal statistics. We tested the hypothesis that short-term facilitation could enable synapses to filter out single spikes and favor bursts of action potentials. We found that the ratio between release probabilities for a burst spike and an isolated spike grows monotonically with increasing number of spikes per burst, and with increasing interval between isolated spikes. Burst detection is optimal when the facilitation time constant matches the average burst duration. Using fractal-like spike patterns characterized by long-term power-law temporal correlations and similar to those seen in sensory neurons, we found that facilitation increases correlation at short time scales. In contrast, depression leads to a dramatic reduction in temporal correlations at all time scales, and to a flat ('whitened') power spectrum, thereby decorrelating natural input signals. PMID- 11053235 TI - Control of the eukaryotic cell cycle by MAP kinase signaling pathways. AB - In an often rapidly changing environment, cells must adapt by monitoring and reacting quickly to extracellular stimuli detected by membrane-bound receptors and proteins. Reversible phosphorylation of intracellular regulatory proteins has emerged as a crucial mechanism effecting the transmission and modulation of such signals and is determined by the relative activities of protein kinases and phosphatases within the cell. These are often arranged into complex signaling networks that may function independently or be subject to cross-regulation. Recently, genetic and biochemical analyses have identified the universally conserved mitogen-activated protein (MAP) kinase cascade as one of the most ubiquitous signal transduction systems. This pathway is activated after a variety of cellular stimuli and regulates numerous physiological processes, particularly the cell division cycle. Progression through the cell cycle is critically dependent on the presence of environmental growth factors and stress stimuli, and failure to correctly integrate such signals into the cell cycle machinery can lead to the accumulation of genetic damage and genomic instability characteristic of cancer cells. Here we focus on the MAP kinase cascade and discuss the molecular mechanisms by which these extensively studied signaling pathways influence cell growth and proliferation. PMID- 11053236 TI - Transgenic study of energy homeostasis equation: implications and confounding influences. AB - Recently novel molecular mediators and regulatory pathways for feeding and body weight regulation have been identified in the brain and the periphery. Mice lacking or overexpressing these mediators or receptors have been produced by molecular genetic techniques, and observations on mutant mice have shed new light on the role of each element in the homeostatic loop of body weight regulation. However, the interpretation of the phenotype is under the potential influence of developmental compensation and other genetic and environmental confounds. Specific alterations of the mediators and the consequences of the altered expression patterns are reviewed here and discussed in the context of their functions as suggested from conventional pharmacological studies. Advanced gene targeting strategies in which genes can be turned on or off at desired tissues and times would undoubtedly lead to a better understanding of the highly integrated and redundant systems for energy homeostasis equation. PMID- 11053237 TI - Subcellular localization of presenilins during mouse preimplantation development. AB - The genes defective in familial Alzheimer's disease encode the proteins presenilin 1 and 2 (PS1 and 2). Expression of presenilins (PSs) and their proteolytic processing are regulated during neuronal development. Even though these proteins are detected and regulated mainly in Golgi and endoplasmic reticulum, their subcellular distribution during the development is not known. The present study aimed to investigate the localization of PSs and their role during early developmental stage using mouse embryo model. At preimplantation stage, PSs were detected not only in cytoplasm, but also in the nucleus from oocyte to 2.5 dpc (day postcoitum), then disappeared in the nucleus at blastocyst stage (3.5 dpc). Antisense against PS1 and PS2 decreased the transition to blastocyst stage, whereas each antisense alone had no effect. Treatment with lactacystin (26S proteosome inhibitor), which arrest cell cycle at M phase, redistributed PSs into centrosome-kinetochore microtubule. PS2 overexpression in HEK 293 cell arrested cell cycle at S phase. These data suggest that PSs play key roles in cell division and differentiation during early development. PMID- 11053238 TI - Allergen mimotopes for 3-dimensional epitope search and induction of antibodies inhibiting human IgE. AB - There is no definite information available on the structural characteristics of IgE binding epitopes on allergenic molecules, although it is widely accepted that most of them are conformational. In the current study we aimed to characterize the IgE epitope of Bet v 1, the major birch pollen allergen, by the application of phage display peptide libraries. We purified IgE specific for Bet v 1 from allergic patients' sera to select mimotopes representing artificial IgE epitopes by biopanning of phage libraries. By linear alignment, it was not possible to attribute mimotope sequences to the primary structure of Bet v 1. We developed a computer-aided, 3-dimensional coarse-grained epitope search. The 3-dimensional search, followed by statistical analysis, revealed an exposed area on the Bet v 1 molecule (located between residues 9-22 and 104-123) as the IgE binding structure. The IgE epitope was located at a 30 A distance from a previously described IgG epitope and the respective mimotope, designated Bet mim E. Such mimotopes could potentially be used for the induction of IgG capable of interfering with the IgE/allergen interaction. To test this hypothesis, we immunized BALB/c mice with the phage-displayed Bet mim E. Immunizations resulted in the induction of Bet v 1-specific IgG, which was able to block the IgE binding to Bet v 1 in vitro. Based on these observations, we propose that immunotherapy with IgE mimotopes generated by biopannings result in formation of blocking IgG. We conclude that mimotope immunotherapy may represent a new and promising concept for treatment of type I allergic disease. PMID- 11053239 TI - Na+/H+ exchanger-dependent intracellular alkalinization is an early event in malignant transformation and plays an essential role in the development of subsequent transformation-associated phenotypes. AB - In this study we investigate the mechanism of intracellular pH change and its role in malignant transformation using the E7 oncogene of human papillomavirus type 16 (HPV16). Infecting NIH3T3 cells with recombinant retroviruses expressing the HPV16 E7 or a transformation deficient mutant we show that alkalinization is transformation specific. In NIH3T3 cells in which transformation can be turned on and followed by induction of the HPV16 E7 oncogene expression, we demonstrate that cytoplasmic alkalinization is an early event and was driven by stimulation of Na+/H+ exchanger activity via an increase in the affinity of the intracellular NHE-1 proton regulatory site. Annulment of the E7-induced cytoplasmic alkalinization by specific inhibition of the NHE-1, acidification of culture medium, or clamping the pHi to nontransformed levels prevented the development of later transformed phenotypes such as increased growth rate, serum-independent growth, anchorage-independent growth, and glycolytic metabolism. These findings were verified in human keratinocytes (HPKIA), the natural host of HPV. Results from both NIH3T3 and HPKIA cells show that alkalinization acts on pathways that are independent of the E2F-mediated transcriptional activation of cell cycle regulator genes. Moreover, we show that the transformation-dependent increase in proliferation is independent of the concomitant stimulation of glycolysis. Finally, treatment of nude mice with the specific inhibitor of NHE-1, DMA, delayed the development of HPV16-keratinocyte tumors. Our data confirm that activation of the NHE-1 and resulting cellular alkalinization is a key mechanism in oncogenic transformation and is necessary for the development and maintenance of the transformed phenotype. PMID- 11053240 TI - Acceleration of phosphatidylcholine synthesis and breakdown by inhibitors of mitochondrial function in neuronal cells: a model of the membrane defect of Alzheimer's disease. AB - Brain cells in Alzheimer's disease (AD) exhibit a membrane defect characterized by accelerated phospholipid turnover. The mechanism responsible for this defect remains unknown. Recent studies indicate that impairment of mitochondrial function is frequently observed in AD and may be responsible for certain aspects of its pathophysiology. We show that when PC12 cells are exposed to inhibitors of mitochondrial bioenergetics, the turnover of their major membrane phospholipid, phosphatidylcholine, is accelerated, producing a pattern of metabolic changes that mimics that observed in brains of AD patients. Abnormalities of mitochondrial function may therefore underlie the membrane defect in AD. PMID- 11053241 TI - Disruption of lens fiber cell architecture in mice expressing a chimeric AQP0-LTR protein. AB - Aquaporin-0 (AQP0) is the major intrinsic protein of lens fiber cells and the founder member of the water channel gene family. Here we show that disruption of the AQP0 gene by an early transposon (ETn) element results in expression of a chimeric protein, comprised of approximately 75% AQP0 and approximately 25% ETn long terminal repeat (LTR) sequence, in the cataract Fraser (CatFr) mouse lens. Immunoblot analysis showed that mutant AQP0-LTR was similar in mass to wild-type AQP0. However, immunofluorescence microscopy revealed that AQP0-LTR was localized to intracellular membranes rather than to plasma membranes of lens fiber cells. Heterozygous CatFr lenses were similar in size to wild-type but displayed abnormal regions of translucence and light scattering. Scanning electron microscopy further revealed that mature fiber cells within the core of the heterozygous CatFr lens failed to stratify into uniform, concentric growth shells, suggesting that the AQP0 water channel facilitates the development of the unique cellular architecture of the crystalline lens. PMID- 11053242 TI - Impaired vascular contractility and blood pressure homeostasis in the smooth muscle alpha-actin null mouse. AB - The smooth muscle (SM) alpha-actin gene activated during the early stages of embryonic cardiovascular development is switched off in late stage heart tissue and replaced by cardiac and skeletal alpha-actins. SM alpha-actin also appears during vascular development, but becomes the single most abundant protein in adult vascular smooth muscle cells. Tissue-specific expression of SM alpha-actin is thought to be required for the principal force-generating capacity of the vascular smooth muscle cell. We wanted to determine whether SM alpha-actin gene expression actually relates to an actin isoform's function. Analysis of SM alpha actin null mice indicated that SM alpha-actin is not required for the formation of the cardiovascular system. Also, SM alpha-actin null mice appeared to have no difficulty feeding or reproducing. Survival in the absence of SM alpha-actin may result from other actin isoforms partially substituting for this isoform. In fact, skeletal alpha-actin gene, an actin isoform not usually expressed in vascular smooth muscle, was activated in the aortas of these SM alpha-actin null mice. However, even with a modest increase in skeletal alpha-actin activity, highly compromised vascular contractility, tone, and blood flow were detected in SM alpha-actin-defective mice. This study supports the concept that SM alpha actin has a central role in regulating vascular contractility and blood pressure homeostasis, but is not required for the formation of the cardiovascular system. PMID- 11053243 TI - Progesterone facilitates chromosome instability (aneuploidy) in p53 null normal mammary epithelial cells. AB - Mammary epithelial cells from p53 null mice have been shown recently to exhibit an increased risk for tumor development. Hormonal stimulation markedly increased tumor development in p53 null mammary cells. Here we demonstrate that mammary tumors arising in p53 null mammary cells are highly aneuploid, with greater than 70% of the tumor cells containing altered chromosome number and a mean chromosome number of 56. Normal mammary cells of p53 null genotype and aged less than 14 wk do not exhibit aneuploidy in primary cell culture. Significantly, the hormone progesterone, but not estrogen, increases the incidence of aneuploidy in morphologically normal p53 null mammary epithelial cells. Such cells exhibited 40% aneuploidy and a mean chromosome number of 54. The increase in aneuploidy measured in p53 null tumor cells or hormonally stimulated normal p53 null cells was not accompanied by centrosome amplification. These results suggest that normal levels of progesterone can facilitate chromosomal instability in the absence of the tumor suppressor gene, p53. The results support the emerging hypothesis based both on human epidemiological and animal model studies that progesterone markedly enhances mammary tumorigenesis. PMID- 11053244 TI - Biodistribution of adenoviral vector to nontarget tissues after local in vivo gene transfer to arterial wall using intravascular and periadventitial gene delivery methods. AB - Expression of transgene other than in the target tissue may cause side effects and safety problems in gene therapy. We analyzed biodistribution of transgene expression after intravascular and periadventitial gene delivery methods using the first generation nuclear-targeted lacZ adenovirus. RT-PCR and X-Gal stainings were used to study transgene expression 14 days after the gene transfer. After intravascular catheter-mediated gene transfer to rabbit aorta mimicking angioplasty procedure, the target vessel showed 1.1% +/- 0. 5 gene transfer efficiency. Other tissues showed varying lacZ gene expression indicating a systemic leakage of the vector with the highest transfection efficiency in hepatocytes (0.7% +/- 0.5). X-Gal staining of blood cells 24 h after the intravascular gene transfer indicated that a significant portion (1.8% +/- 0.8) of circulating monocytes was transfected. X-Gal-positive cells were also found in testis. After periadventitial gene transfer using a closed silicon capsule placed around the artery, 0.1% +/- 0.1 lacZ-positive cells were detected in the artery wall. Positive cells were also found in the liver and testis (<0.01%), indicating that the virus escapes even from the periadventitial space, although less extensively than during the intravascular application. We conclude that catheter mediated intravascular and, to a lesser extent, periadventitial gene transfer lead to leakage of adenovirus to systemic circulation, followed by expression of the transgene in several tissues. Possible consequences of the ectopic expression of the transgene should be evaluated in gene therapy trials even if local gene delivery methods are used. PMID- 11053245 TI - The role of differential activation of p38-mitogen-activated protein kinase in preconditioned ventricular myocytes. AB - Activation of protein kinase C (PKC) and more recently mitogen-activated protein kinases (MAPKs) have been associated with the cardioprotective effect of ischemic preconditioning. We examined the interplay between these kinases in a characterized model of ischemic preconditioning in cultured rat neonatal ventricular cardiocytes where ectopic expression of active PKC-delta results in protection. Two members of the MAPK family, p38 and p42/44, were activated transiently during preconditioning by brief simulated ischemia/reoxygenation. Overexpression of active PKC-delta, rather than augmenting, completely abolished this activation. We therefore determined whether a similar process occurred during lethal prolonged simulated ischemia. In contrast to ischemia, brief, lethal-simulated ischemia activated only p38 (2.8+/-0.45 vs. basal, P<0.01), which was attenuated by expression of active PKC-delta or by preconditioning (0.48+/-0.1 vs. ischemia, P<0.01). To determine whether reduced p38 activation was the cause or an effect of protection, we used SB203580, a p38 inhibitor. SB203580 reduced ischemic injury (CK release 38.0+/-3.1%, LDH release 77.3+/ 4.0%, and MTT bioreduction 127.1+/-4.8% of control, n=20, P<0.05). To determine whether p38 activation was isoform selective, myocytes were infected with adenoviruses encoding wild-type p38alpha or p38beta. Transfected p38alpha and beta show differential activation (P<0.001) during sustained simulated ischemia, with p38alpha remaining activated (1.48+/-0.36 vs. basal) but p38beta deactivated (0.36+/-0.1 vs. basal, P<0.01). Prior preconditioning prevented the activation of p38alpha (0.65+/-0.11 vs. ischemia, P<0.05). Moreover, cells expressing a dominant negative p38alpha, which prevented ischemic p38 activation, were resistant to lethal simulated ischemia (CK release 82.9+/-3.9% and MTT bioreduction 130.2+/-6.5% of control, n=8, P<0.05). Thus, inhibition of p38alpha activation during ischemia reduces injury and may contribute to preconditioning induced cardioprotection in this model. PMID- 11053246 TI - Quantitative trait loci modulate neutrophil infiltration in the liver during LPS induced inflammation. AB - A crucial aspect of the inflammatory response is the recruitment of activated neutrophils (PMN) to the site of damage. Lytic enzymes and oxygen radicals released by PMN are important in clearing an infection or cellular debris, but can also produce host tissue damage. Failure to properly regulate the inflammatory response contributes to a variety of human diseases like sepsis and multiple organ dysfunction syndrome, the leading cause of morbidity and mortality in surgical intensive care units. Many aspects of human disease pathology, including hepatic PMN infiltration, can be recapitulated in mice using an endotoxic shock model. Six quantitative trait loci that predispose to high infiltration of PMN in hepatic sinusoids after high-dose endotoxin administration were provisionally identified. Two of these loci, Hpi1 and Hpi2 on mouse chromosomes 5 and 13, were mapped to the significant and highly significant level using a low-resolution genome scan on 122 intercross animals. These loci interact epistatically to produce a high degree of PMN infiltration. Intercross and recombinant inbred strain mice with a specific genotype at these loci always had a high infiltration response, indicating that genotype analysis at just these two loci can accurately predict a high PMN infiltration response. Genetic predisposition to the degree of PMN infiltration in the inflammatory response in mice suggests that analogous genetic mechanisms occur in human beings that could be used for diagnostic purposes. PMID- 11053247 TI - Sphingosine 1-phosphate released from platelets during clotting accounts for the potent endothelial cell chemotactic activity of blood serum and provides a novel link between hemostasis and angiogenesis. AB - Recent studies have identified factors responsible for angiogenesis within developing tumors, but mediators of vessel formation at sites of trauma, injury, and wound healing are not clearly established. Here we show that sphingosine 1 phosphate (S1P) released by platelets during blood clotting is a potent, specific, and selective endothelial cell chemoattractant that accounts for most of the strong endothelial cell chemotactic activity of blood serum, an activity that is markedly diminished in plasma. Preincubation of endothelial cells with pertussis toxin inhibited this effect of S1P, demonstrating the involvement of a Galphai-coupled receptor. After S1P-induced migration, endothelial cells proliferated avidly and differentiated forming multicellular structures suggestive of early blood vessel formation. S1P was strikingly effective in enhancing the ability of fibroblast growth factor to induce angiogenesis in the avascular mouse cornea. Our results show that blood coagulation initiates endothelial cell angiogenic responses through the release of S1P, a potent endothelial cell chemoattractant that exerts its effects by activating a receptor dependent process. PMID- 11053248 TI - H2O2-induced block of glycolysis as an active ADP-ribosylation reaction protecting cells from apoptosis. AB - H2O2 treatment on U937 cells leads to the block of glycolytic flux and the inactivation of glyceraldehyde-3-phosphate-dehydrogenase by a posttranslational modification (possibly ADP-ribosylation). Glycolysis spontaneously reactivates after 2 h of recovery from oxidative stress; thereafter cells begin to undergo apoptosis. The specific ADP-ribosylation inhibitor 3-aminobenzamide inhibits the stress-induced inactivation of glyceraldehyde-3-phosphate-dehydrogenase and the block of glycolysis; concomitantly, it anticipates and increases apoptosis. Exogenous block of glycolysis (i.e., by culture in glucose-free medium or with glucose analogs or after NAD depletion), turns the transient block into a stable one: this results in protection from apoptosis, even when downstream cell metabolism is kept active by the addition of pyruvate. All this evidence indicates that the stress-induced block of glycolysis is not the result of a passive oxidative damage, but rather an active cell reaction programmed via ADP ribosylation for cell self-defense. PMID- 11053249 TI - Expression of endothelin 1 and endothelin A receptor in HPV-associated cervical carcinoma: new potential targets for anticancer therapy. AB - Human papillomaviruses (HPV) are associated with cervical cancer and interact with growth factors that may enhance malignant transformation of cervical carcinoma cells. Endothelin-1 (ET-1) is released from HPV transfected keratinocytes and induces increased growth response in these cell lines in comparison with normal cells. In the present study several cervical carcinoma cell lines have been analyzed to investigate the expression of ET-1 and its receptors as well as their involvement in tumor growth. All HPV-positive cancer cells secreted ET-1 and expressed mRNA for ET-1 and its receptors, whereas a HPV negative carcinoma cell line expressed only the ETBR mRNA and didn't secrete ET 1. Binding studies showed that HPV-associated cells expressed an increased number of functional ETAR. ET-1 stimulated a marked dose-dependent increase in [3H] thymidine incorporation with respect to the normal cells whereas ET-3 and ETBR agonists had no effect. In HPV-positive cancer cells, a specific antagonist of ETAR inhibited the proliferation induced by ET-1 and substantially reduced the basal growth rate of unstimulated cervical tumor cells, whereas the ETBR antagonist had no effect. These results demonstrate that ET-1 participates in the progression of neoplastic growth in HPV-associated carcinoma, in which ETAR are increased and could be targeted for antitumor therapy. PMID- 11053250 TI - Lyn and syk tyrosine kinases are not activated in B-lineage lymphoid cells exposed to low-energy electromagnetic fields. AB - Exposure of B-lineage lymphoid cells to a 100 microT 60 Hz AC magnetic field has been reported to stimulate the rapid activation of Lyn and Syk tyrosine kinases and the induction of protein tyrosine phosphorylation. These findings are significant because of the critical role played by these B cell signaling events in the control of growth and differentiation, and therefore the potential of electromagnetic field (EMF) exposure to induce cancer. We report the first study carried out with the aim of reproducing the reported EMF effects on Lyn and Syk tyrosine kinases. The system used enabled EMF exposure conditions to be carefully controlled and also allowed experiments to be performed blind. The effects of a 100 microT 60 Hz AC magnetic field on protein tyrosine phosphorylation and on Lyn and Syk tyrosine kinase activities were investigated in Nalm-6 and DT40 B cells in the absence and presence of a 46 microT DC magnetic field. However, no significant effects of low-energy electromagnetic fields on tyrosine kinase activities or protein phosphorylation were observed. PMID- 11053251 TI - A gain of function p53 mutant promotes both genomic instability and cell survival in a novel p53-null mammary epithelial cell model. AB - Approximately 40% of human breast cancers contain alterations in the tumor suppressor p53. The p53 172R-H gain-of-function mutant (equivalent to the common 175R-H human breast cancer mutant) has been shown to promote aneuploidy and tumorigenesis in the mammary gland in transgenic mice and may affect genomic stability in part by causing centrosome abnormalities. The precise mechanism of action of these gain-of-function mutants is not well understood, and has been studied primarily in fibroblast cell lines. A novel p53-null mouse mammary epithelial cell line developed from p53-null mice has been used in adenovirus mediated transient transfection experiments to study the properties of this p53 mutant. Marked centrosome amplification and an increased frequency of aberrant mitoses were observed within 72 h of introduction of p53 172R-H. However, few cells with aberrant centrosome numbers were observed in cells stably expressing the p53 172R-H mutant. Furthermore, stable expression of this p53 mutant reduced both basal and DNA damage-induced apoptosis. This result may be mediated in part through abrogation of p73 function. The p53 172R-H mutant, therefore, appears to influence tumorigenesis at the molecular level in two distinct ways: promoting the development of aneuploidy in cells while also altering their apoptotic response after DNA damage. PMID- 11053252 TI - Uncoupling of betaIIPKC from its targeting protein RACK1 in response to ethanol in cultured cells and mouse brain. AB - Protein kinase C (PKC) is involved in many neuroadaptive responses to ethanol in the nervous system. PKC activation results in translocation of the enzyme from one intracellular site to another. Compartmentalization of PKC isozymes is regulated by targeting proteins such as receptors for activated C kinase (RACKs). It is possible, therefore, that ethanol-induced changes in the function and compartmentalization of PKC isozymes could be due to changes in PKC targeting proteins. Here we study the response of the targeting protein RACK1 and its corresponding kinase betaIIPKC to ethanol, and propose a novel mechanism to explain how ethanol modulates signaling cascades. In cultured cells, ethanol induces movement of RACK1 to the nucleus without affecting the compartmentalization of betaIIPKC. Ethanol also inhibits betaIIPKC translocation in response to activation. These results suggest that ethanol inhibition of betaIIPKC translocation is due to miscompartmentalization of the targeting protein RACK1. Similar events occurred in mouse brain. In vivo exposure to ethanol caused RACK1 to localize to nuclei in specific brain regions, but did not affect the compartmentalization of betaIIPKC. Thus, some of the cellular and neuroadaptive responses to ethanol may be related to ethanol-induced movement of RACK1 to the nucleus, thereby preventing the translocation and corresponding function of betaIIPKC. PMID- 11053253 TI - De novo-synthesized ceramide signals apoptosis in astrocytes via extracellular signal-regulated kinase. AB - Recent observations support the importance of ceramide synthesis de novo in the induction of apoptosis. However, the downstream targets of de novo-synthesized ceramide are unknown. Here we show that palmitate incorporated into ceramide and induced apoptotic DNA fragmentation in astrocytes. These effects of palmitate were exacerbated when fatty acid breakdown was uncoupled and were not evident in neurons, which show a very low capacity to take up and metabolize palmitate. Palmitate-induced apoptosis of astrocytes was prevented by L-cycloserine and fumonisin B1, two inhibitors of ceramide synthesis de novo, and by PD098059, an inhibitor of the extracellular signal-regulated kinase (ERK) cascade. Accordingly, palmitate activated ERK by a process that was dependent on ceramide synthesis de novo and Raf-1, but independent of kinase suppressor of Ras. Other potential targets of ceramide in the control of cell fate, namely, c-Jun amino terminal kinase, p38 mitogen-activated protein kinase, and protein kinase B, were not significantly affected in astrocytes exposed to palmitate. Results show that the Raf-1/ERK cascade is the selective downstream target of de novo-synthesized ceramide in the induction of apoptosis in astrocytes and also highlight the importance of ceramide synthesis de novo in apoptosis of astrocytes, which might have pathophysiological relevance. PMID- 11053254 TI - Human respiratory syncytial virus vaccine antigen produced in plants. AB - Human respiratory syncytial virus (RSV) is the primary cause of respiratory infection in infants worldwide. Currently there is no available vaccine, although studies in animal models have demonstrated protective immunity induced by an epitope of the RSV G-protein representing amino acids 174-187. Two peptides containing amino acids 174-187 of the G-protein of the human RSV A2 strain (NF1 RSV/172-187 and NF2-RSV/170-191) were separately engineered as translational fusions with the alfalfa mosaic virus coat protein and individually expressed in Nicotiana tabacum cv. Samsun NN plants through virus infection. RSV G-protein peptides were expressed in infected plant tissues at significant levels within 2 wk of inoculation and purified as part of recombinant alfalfa mosaic virions. BALB/c mice immunized intraperitoneally with three doses of the purified recombinant viruses showed high levels of serum antibody specific for RSV G protein and were protected against infection with RSV Long strain. PMID- 11053255 TI - Leptin promotes invasiveness of kidney and colonic epithelial cells via phosphoinositide 3-kinase-, rho-, and rac-dependent signaling pathways. AB - Leptin plays a key role regulating food intake, body weight and fat mass. These critical parameters are associated with an increased risk for digestive and mammary gland cancer in the Western population. Here we determined whether leptin contributes to the invasive phenotype of colonic and kidney epithelial cells at various stages of the neoplastic progression. First, leptin potently (EC50 = 10 30 ng/ml) induces invasion of collagen gels by premalignant familial adenomatous colonic cells PC/AA/C1 and nontumorigenic MDCK kidney epithelial cells, their src transformed counterparts, and the human adenocarcinoma colonic cells LoVo and HCT 8/S11. Leptin and its Ob-Rb receptors were consistently identified by RT-PCR and immunoblotting in these cell lines, as well as in human colonic epithelial crypts, polyps, colonic tumor resections, and adjacent mucosa. Leptin-induced invasion was effectively blocked by pharmacological inhibitors of several downstream signaling pathways involved in cell transformation, namely, JAK2 tyrosine kinase (AG490), phosphoinositide PI3'-kinase (wortmannin and LY294002), mTOR kinase (rapamycin), and protein kinases C (GF109203X, Go6976). Accordingly, leptin induces transient elevation of the PI3'-kinase lipid products in JAK2 immunoprecipitates prepared from parental MDCK cells. The leptin effect on invasion was potentiated by the activated form of the small GTPase RhoA and was abrogated by dominant negative mutants of RhoA, Rac1, and the p110alpha of PI3' K. Our data indicate that leptin may exert a local and beneficial effect on migration of normal colonic epithelial cells and reparation of the inflamed or wounded digestive mucosa. We also emphasize a new role for leptin, linking the nutritional and body fat status to digestive cancer susceptibility by stimulating the invasive capacity of colonic epithelial cells at early stages of neoplasia. This finding has potential clinical implications for colon cancer progression and management of obesity. PMID- 11053256 TI - Thioguanine substitution alters DNA cleavage mediated by topoisomerase II. AB - Thiopurines and topoisomerase II-targeted drugs (e.g., etoposide) are widely used anticancer drugs. However, topoisomerase II-targeted drugs can cause acute myeloid leukemia, with the risk of this secondary leukemia linked to a genetic defect in thiopurine catabolism. Chronic thiopurines result in thioguanine substitution in DNA. The effect of these substitutions on DNA topoisomerase II activity is not known. Our goal was to determine whether deoxythioguanosine substitution alters DNA cleavage stabilized by human topoisomerase II. We studied four variations of a 40 mer oligonucleotide with a topoisomerase II cleavage site, each with a single deoxythioguanosine in a different position relative to the cleavage site (-1 or +2 in the top and +2 or +4 in the bottom strand). Deoxythioguanosine substitution caused position-dependent quantitative effects on cleavage. With the -1 or +2 top and +2 or +4 bottom substitutions, mean topoisomerase II-induced cleavage was 0.6-, 2.0-, 1.1-, and 3.3-fold that with the wild-type substrate (P=0. 011, < 0.008, 0.51, and < 0.001, respectively). In the presence of 100 microM etoposide, cleavage was enhanced for wild-type and all thioguanosine-modified substrates relative to no etoposide, with the +4 bottom substitution showing greater etoposide-induced cleavage than the wild-type substrate (P=0.015). We conclude that thioguanine incorporation alters the DNA cleavage induced by topoisomerase II in the presence and absence of etoposide, providing new insights to the mechanism of thiopurine effect and on the leukemogenesis of thiopurines, with or without topoisomerase inhibitors. PMID- 11053257 TI - Ca2+-activated Cl- channels can substitute for CFTR in stimulation of pancreatic duct bicarbonate secretion. AB - This study addresses the mechanisms by which a defect in CFTR impairs pancreatic duct bicarbonate secretion in cystic fibrosis. We used control (PANC-1) and CFTR deficient (CFPAC-1; DeltaF508 mutation) cell lines and measured HCO3- extrusion by the rate of recovery of intracellular pH after an alkaline load and recorded whole cell membrane currents using patch clamp techniques. 1) In PANC-1 cells, cAMP causes parallel activation of Cl- channels and of HCO3- extrusion by DIDS sensitive and Na+-independent Cl-/HCO3- exchange, both effects being inhibited by Cl- channel blockers NPPB and glibenclamide. 2) In CFPAC-1 cells, cAMP fails to stimulate Cl-/HCO3- exchange and Cl- channels, except after promoting surface expression of DeltaF508-CFTR by glycerol treatment. Instead, raising intracellular Ca2+ concentration to 1 micromol/l or stimulating purinergic receptors with ATP (10 and 100 micromol/l) leads to parallel activation of Cl- channels and HCO3- extrusion. 3) K+ channel function is required for coupling cAMP- and Ca2+-dependent Cl- channel activation to effective stimulation of Cl /HCO3- exchange in control and CF cells, respectively. It is concluded that stimulation of pancreatic duct bicarbonate secretion via Cl-/HCO3- exchange is directly correlated to activation of apical membrane Cl- channels. Reduced bicarbonate secretion in cystic fibrosis results from defective cAMP-activated Cl channels. This defect is partially compensated for by an increased sensitivity of CF cells to purinergic stimulation and by alternative activation of Ca2+ dependent Cl- channels, mechanisms of interest with respect to possible treatment of cystic fibrosis and of related chronic pancreatic diseases. PMID- 11053258 TI - C-peptide inhibits leukocyte-endothelium interaction in the microcirculation during acute endothelial dysfunction. AB - C-peptide is a cleavage product that comes from processing proinsulin to insulin that induces nitric oxide (NO) -mediated vasodilation. NO modulates leukocyte endothelium interaction. We hypothesized that C-peptide might inhibit leukocyte endothelium interaction via increased release of endothelial NO. Using intravital microscopy of the rat mesentery, we measured leukocyte-endothelium interactions after administration of C-peptide to the rat. Superfusion of the rat mesentery with either thrombin or L-NAME consistently and significantly increased the number of rolling, adhering, and transmigrated leukocytes. C-peptide significantly attenuated either thrombin- or L-NAME-induced leukocyte-endothelium interactions in rat mesenteric venules. A control scrambled sequence of C-peptide characterized by the same amino acid composition in a randomized sequence failed to inhibit leukocyte-endothelium interactions. These effects of C-peptide were associated with decreased surface expression of the cell adhesion molecules P selectin and ICAM-1 on the microvascular endothelium. Endothelial nitric oxide synthase (eNOS) mRNA levels were increased in rats injected with C-peptide. This enhanced eNOS expression was associated with a marked increase in basal NO release from the aorta of C-peptide-treated rats. We conclude that C-peptide is a potent inhibitor of leukocyte-endothelium interaction and that this effect is specifically related to inhibition of endothelial cell adhesion molecules via maintenance of NO release from the vascular endothelium. PMID- 11053259 TI - Phylogenomic analysis of 16S rRNA:(guanine-N2) methyltransferases suggests new family members and reveals highly conserved motifs and a domain structure similar to other nucleic acid amino-methyltransferases. AB - The sequences of known Escherichia coli 16S rRNA:m2G1207 methyltransferase (MTase) RsmC and hypothetical 16S rRNA:m2G966 MTase encoded by the ygjo open reading frame were used to carry out a database search of other putative m2G generating enzymes in finished and unfinished genomic sequences. Sequence comparison and phylogenetic analysis of 21 close homologs of RsmC and YgjO revealed the presence of the third paralogous lineage in E. coli and other gamma Proteobacteria, which might correspond to the subfamily of MTases specific for G1516 in 16S rRNA. In addition, the comparative sequence analysis supported by sequence/structure threading suggests that rRNA:m2G MTases are very closely related to RNA and DNA:m6A MTases and that these two enzyme families share common architecture of the active site and presumably a similar mechanism of methyl group transfer onto the exocyclic amino group of their target bases. PMID- 11053261 TI - IkappaB kinase alpha is essential for development of the mammalian cornea and conjunctiva. AB - PURPOSE: To determine the requirement of IkappaB kinase alpha (Ikkalpha) for differentiation of the mammalian cornea and conjunctiva. METHODS: Newborn mice or surgically removed embryonic day (E)18 to E19 fetuses of wild-type and IKK:alpha( /-) mice were analyzed by light microscopy and electron microscopy or immunocytochemistry using anti-keratin (K)12, K4, K5, IkappaB, or nuclear factor (NF)-kappaB (p50) antibody. RESULTS: In the IKKalpha(-/-) eyes, the epithelium of the cornea and the conjunctiva consisted of poorly differentiated cells with round nuclei. K5 was much stronger in the conjunctiva of the IKKalpha(-/-) mice. Expression of K12 in the cornea and K4 in the conjunctiva was impaired in the IKKalpha(-/-) mice. IkappaB expression was low in epithelium of the cornea and conjunctiva of the wild type mice but was very strong in that of the IKKalpha(-/ ) mice. During normal development of the conjunctiva, nuclear localization of p50 was seen in areas where basal undifferentiated cells give rise to differentiated cell types, marked by expression of cK4. However, in the IKK++alpha(-/-) tissues, no nuclear p50 staining was detected. CONCLUSIONS: IKKalpha is specifically required for formation of cornea and conjunctiva. This function may be exerted through an effect on NF-kappaB activity. PMID- 11053260 TI - British Society of Gastroenterology guidelines for the management of the irritable bowel syndrome. PMID- 11053262 TI - Decreased GlcNAc 6-O-sulfotransferase activity in the cornea with macular corneal dystrophy. AB - PURPOSE: Macular corneal dystrophy (MCD) is an autosomal recessive inherited disorder that is accompanied by corneal opacity. Explants from MCD-affected corneas have been reported to synthesize low-sulfated KS, suggesting that sulfate groups attached to KS may play critical roles in maintaining corneal transparency. To clear the biosynthetic defect in the MCD cornea, sulfotransferase activities were determined that are presumably involved in the biosynthesis of KS: galactose-6-sulfotransferase (Gal6ST) activity and N acetylglucosamine 6-O-sulfotransferase (GlcNAc6ST) activity. METHODS: Gal6ST and GlcNAc6ST activities, which were contained in the corneal extracts from corneas affected by MCD and keratoconus and from normal control corneas, were determined by measuring the transfer of (35)SO(4) from [(35)S]3'-phosphoadenosine 5' phosphosulfate into the Gal residue of partially desulfated KS and the nonreducing terminal GlcNAc residue of GlcNAcbeta1-3Galbeta1-4GlcNAc (oligo A), respectively. RESULTS: The level of Gal6ST activity in corneal extracts from eyes with MCD, which was measured by using partially desulfated KS as an acceptor, was nearly equal to that in eyes with keratoconus and normal control eyes. In contrast, GlcNAc6ST activity in the extracts from MCD-affected corneas, which was measured by using oligo A as an acceptor, was much lower than in those in corneas with keratoconus and in normal control corneas. CONCLUSIONS: The decrease in GlcNAc6ST activity in the cornea with MCD may result in the occurrence of low- or nonsulfated KS and thereby cause corneal opacity. PMID- 11053263 TI - Characterization of gene expression in human trabecular meshwork using single pass sequencing of 1060 clones. AB - PURPOSE: To study the gene expression profile of the human trabecular meshwork (HTM). METHODS: A polymerase chain reaction (PCR)-amplified cDNA library was constructed using RNA from the TM of a 67-year-old normal, perfused human eye. A total of 1060 clones were randomly selected for sequencing of one end. These sequences were searched against nonredundant GenBank and dbEST databases for similarity comparison by using a FASTA file and the BLASTcl3 program. Relative expression patterns of those clones that matched other expressed sequence tags (ESTs) were determined using the National Center for Biotechnology Information (NCBI) Unique Human Gene Sequence Collection (UniGene) database. RESULTS: Of the 1060 clones analyzed, 519 (48.9%) had sequences identical with known genes, 125 (11.8%) matched ESTs, and 189 (17.8%) did not match any database sequences. Of the remaining clones, 31 (3%) corresponded to mitochondrial transcripts and 196 (18.5%) to repetitive and noninformative sequences. It is notable that some of the genes highly represented in this library are not ubiquitously expressed in other tissues, which suggests a potentially important role in the HTM. As evidence for the presence of true novel genes in the library, one of the clones was fully sequenced. This clone comprised a complete open reading frame of 966 nucleotides, and its deduced amino acid sequence corresponded to a protein 33% similar to the MAS-related G-protein-coupled receptor. CONCLUSIONS: The identification of the more highly expressed genes in HTM and the discovery of novel genes expressed in this tissue provides basic information for further research on the physiology of the TM and for the identification of glaucoma candidate genes. PMID- 11053264 TI - Circadian-dependent retinal light damage in rats. AB - PURPOSE: To determine the relative susceptibility of rats to retinal light damage at different times of the day or night. METHODS: Rats maintained in a dim cyclic light or dark environment were exposed to a single dose of intense green light beginning at various times. Normally, light exposures were for 8 or 3 hours, respectively, although longer and shorter periods were also used. Some animals were treated with the synthetic antioxidant dimethylthiourea (DMTU) before or after the onset of light. The extent of visual cell loss was estimated from measurements of rhodopsin and retinal DNA levels 2 weeks after light treatment. The time course of retinal DNA fragmentation, and the expression profiles of heme oxygenase-1 (HO-1) and interphotoreceptor retinol binding protein (IRBP) were determined 1 to 2 days after exposure. RESULTS: When dark-adapted, cyclic light reared or dark-reared rats were exposed to intense light during normal nighttime hours (2000-0800) the loss of rhodopsin or photoreceptor cell DNA was approximately twofold greater than that found in rats exposed to light during the day (0800-2000). The relative degree of light damage susceptibility persisted in cyclic light-reared rats after dark adaptation for up to 3 additional days. For rats reared in a reversed light cycle, the light-induced loss of rhodopsin was also reversed. Longer duration light treatments revealed that dim cyclic light reared rats were three- to fourfold more susceptible to light damage at 0100 than at 1700 and that dark-reared animals were approximately twofold more susceptible. Intense light exposure at 0100 resulted in greater retinal DNA fragmentation and the earlier appearance of apoptotic DNA ladders than at 1700. The extent of retinal DNA damage also correlated with an induction of retinal HO-1 mRNA and with a reduction in IRBP transcription. Antioxidant treatment with DMTU was effective in preventing retinal light damage when given before but not after the onset of light. CONCLUSIONS: These results confirm earlier work showing greater retinal light damage in rats exposed at night rather than during the day and extend those findings by demonstrating that a single, relatively short, intense light exposure causes a circadian-dependent, oxidatively induced loss of photoreceptor cells. The light-induced loss of photoreceptor cells is preceded by DNA fragmentation and by alterations in the normal transcriptional events in the retina and within the photoreceptors. The expression profile of an intrinsic retinal factor(s) at the onset of light exposure appears to be important in determining light damage susceptibility. PMID- 11053265 TI - Localization of the pathogenic gene of Behcet's disease by microsatellite analysis of three different populations. AB - PURPOSE: Behcet's disease (BD) is known to be associated with HLA-B51 in many ethnic groups. However, the pathogenic gene responsible for BD is as yet unknown. To localize the critical region of the pathogenic gene, microsatellite markers distributed around the HLA-B gene were investigated. The BD patients studied were of three ethnic origins: Japanese, Greek, or Italian. METHODS: The total group consisted of 172 BD patients, of whom were 95 Japanese, 55 Greek, and 22 Italian. Eight polymorphic microsatellite markers distributed within 1100 kb of the HLA-B gene were analyzed using PCR and subsequent automated fragment detection by fluorescent-based technology. RESULTS: Among the eight markers, allele 348 of the MIB microsatellite was remarkably common in all three BD populations (Japanese, PC: = 0.000014; Greek, PC: = 0. 00047; Italian, PC: = 0.11). However, HLA-B51 was found to be the marker most strongly associated with BD in each population (Japanese, PC: = 0.000000000017; Greek, PC: = 0.00000032; Italian, PC: = 0. 0074). In genotypic differentiation between the patients and controls, only HLA B51 was found to be significantly associated with BD in all three populations. Stratification analysis suggested that significant associations of BD with MICA and other microsatellites resulted from a linkage disequilibrium with HLA-B51. CONCLUSIONS: These results suggest that the pathogenic gene of BD is HLA-B51 itself and not other genes located in the vicinity of HLA-B. PMID- 11053266 TI - Novel locus for autosomal recessive cone-rod dystrophy CORD8 mapping to chromosome 1q12-Q24. AB - PURPOSE: To map the disease locus of a two-generation, consanguineous Pakistani family with autosomal recessive cone-rod dystrophy (arCRD). All affected individuals had night blindness, deterioration of central vision, photophobia, epiphora in bright light, and problems with color distinction. Fundoscopy revealed marked macular degeneration and attenuation of retinal vessels. Mild pigmentary changes were present in the periphery. METHODS: Genomic DNA was amplified across the polymorphic microsatellite poly-CA regions identified by markers. Alleles were assigned to individuals that allowed calculation of LOD scores using the Cyrillic (Cherwell Scientific, Oxford, UK) and MLINK (accessed from ftp://linkage. rockefeller.edu/softeware/linkage/) software programs. The cellular retinoic acid-binding protein 2 (CRABP2), cone transducin alpha-subunit (GNAT2), potassium inwardly rectifying channel, subfamily J, member 10 (KCNJ10), genes were analyzed by heteroduplex analysis and direct sequencing for mutations. RESULTS: A new locus for arCRD (CORD8) has been mapped to chromosome 1q12-q24. A maximum two-point LOD score of 4.22 was obtained with marker D1S2635 at recombination fraction of theta = 0.00. Two critical recombinations in the pedigree positioned this locus to a region flanked by markers D1S457 and D1S2681. A region of homozygosity was observed within the loci D1S442 and D1S2681, giving a probable critical disease interval of 21 cM. Mutation screening of the three candidate genes CRABP2, GNAT2, and KCNJ10 revealed no disease-associated mutations. CONCLUSIONS: The findings therefore suggest that this phenotype maps to a new locus and is due to an as yet uncharacterized gene within the 1q12-q24 chromosomal region. PMID- 11053267 TI - Scleral remodeling during the development of and recovery from axial myopia in the tree shrew. AB - PURPOSE: Recent investigations have suggested that scleral thinning in mammalian eyes with axial myopia is a consequence of the loss of scleral tissue, rather than the redistribution of existing tissue as the eye enlarges. The present study investigated whether further changes in the distribution and metabolism of scleral tissue occur during the process of recovery from axial myopia. Scleral glycosaminoglycan (GAG) synthesis and content as well as scleral dry weight changes were monitored as indicators of remodeling in myopic and recovering tree shrew sclerae. METHODS: Myopia was induced in tree shrews by monocularly depriving them of pattern vision. Some animals then had the occluder removed and were allowed to recover from the induced myopia for periods of 1, 3, 5, 7, and 9 days. Newly synthesized GAGs were radiolabeled in vivo with [(35)S]sulfate. Sulfate incorporation and total GAG content in the sclera was measured through selective precipitation of GAGs from proteinase K digests with alcian blue dye. Dry weights of the sclerae were also determined. Changes in ocular refraction and eye size were monitored using retinoscopy, keratometry, and ultrasonography. RESULTS: Eyes developing myopia showed a significant reduction in scleral GAG synthesis, particularly in the region of the posterior pole (-36% +/- 7%) compared with contralateral control eyes. Scleral dry weight was also significantly reduced in these eyes (-3.7% +/- 1.2%). In recovering eyes, significant changes in GAG synthesis were apparent after 24 hours of recovery. After 3 days of recovery, significantly elevated levels of GAG synthesis were found (+79% +/- 15%), returning to contralateral control eye values after 9 days of recovery. Interocular differences in scleral dry weight were shown to follow a similar pattern to that observed for GAG synthesis. CONCLUSIONS: Active remodeling, resulting in either the loss or replacement of scleral tissue and not passive redistribution of scleral tissue, is associated with changes in eye size during both myopia development and recovery. Regulatory changes in scleral metabolism can be rapidly evoked by a change in visual conditions and the direction of regulation is related to the direction of change in eye size. PMID- 11053268 TI - Attributable risk estimates for cataract to prioritize medical and public health action. AB - PURPOSE: Cataract is the most common cause of blindness in the world. The purpose of this study was to estimate the population attributable risk associated with identified risk factors for cortical, nuclear, and posterior subcapsular (PSC) cataract in a representative sample of the Victorian population aged 40 years and older. METHODS: Cluster, stratified sampling was used and participants were recruited through a household census. At locally established test sites, standardized clinical examinations were performed to assess cataract and personal interviews were conducted to quantify potential risk factors. Multivariate logistic regression was used to determine the independent risk factors associated with the three types of cataract, and the population attributable risk was calculated. RESULTS: A total of 3271 (83% of eligible) of the urban residents and 1473 (92%) rural residents participated. The urban residents ranged in age from 40 to 98 years (mean, 59 years), and 1511 (46%) were men. The rural residents ranged in age from 40 to 103 years (mean, 60 years), and 701 (48%) were men. The overall prevalence of cortical cataract was 12.1% (95% CL 10.5, 13.8), nuclear cataract 12.6% (95% CL 9.61, 15.7), and PSC cataract 4.93% (95% CL 3.68, 6.17). Significant risk factors for cortical cataract included age, female gender, diabetes for greater than 5 years, gout for greater than 20 years, arthritis, myopia, average annual ocular UV-B exposure, and family history of cataract (parents or siblings). Significant risk factors for nuclear cataract included age, female gender, rural residence, age-related maculopathy, diabetes for greater than 5 years, smoker for greater than 30 years, and myopia. The significant risk factors for PSC cataract were age, rural residence, thiazide diuretic use, and myopia. Of the modifiable risk factors, ocular UV-B exposure explains 10% of the cortical cataract in the community, and cigarette smoking accounts for 17% of the nuclear cataract. CONCLUSIONS: Because of the near universal exposure to UV-B in the environment, ocular protection has one of the highest modifiable attributable risks for cortical cataract and would therefore be an ideal target for public health intervention. Quit smoking campaigns can be expanded to incorporate information about the excess cataract in the community associated with long-term smoking. Nonmodifiable risk factors such as age, gender, and long-term medication use have implications for the timely referral and treatment for those at higher risk of cataract. PMID- 11053269 TI - Normal emmetropization in infants with spectacle correction for hyperopia. AB - PURPOSE: The development of emmetropic refraction is known to be under visual control. Does partial spectacle correction of infants' refractive errors, which has been shown to have beneficial effects in reducing strabismus and amblyopia, impede emmetropization? The purpose of the present study was to perform the first longitudinal controlled trial to investigate this question in human subjects. METHODS: Children identified as having significant hyperopia in a population screening program at age 8 to 9 months were assigned to treated (partial spectacle correction) or untreated groups. A control group of infants with no significant refractive errors at screening was also recruited. Measurements of retinoscopic refraction under cycloplegia were taken at 4- to 6-month intervals up to the age of 36 months, and changes in refraction of 148 subjects were analyzed longitudinally. RESULTS: Refractive error decreased toward low hyperopic values between 9 and 36 months in both hyperopic groups. By 36 months, this reduction of hyperopia showed no overall difference between children who were treated with partial spectacle correction and those who were not. Despite the improvement, both hyperopic groups' mean refractive error at 36 months remained higher than that of the control group. When infants in all three groups were considered together, the rate of reduction of refractive error was, on average, a linear function of the initial level of hyperopia. CONCLUSIONS: The benefits of spectacle correction for infants with hyperopia can be achieved without impairing the normal developmental regulation of refraction. PMID- 11053270 TI - Androgen influence on the meibomian gland. AB - PURPOSE: The hypothesis in the study was that androgens control meibomian gland function, regulate the quality and/or quantity of lipids produced by this tissue, and promote the formation of the tear film's lipid layer. To test this hypothesis, a study was conducted to determine whether androgen receptor protein exists in the epithelial cell nuclei of rat meibomian glands and, in addition, whether androgen deficiency and/or treatment influences the gross morphology, neutral lipid content, and fatty acid profile of the rabbit meibomian gland, as well as the appearance of the tear film lipid layer. METHODS: Rat lids were obtained and processed for immunohistochemistry. Meibomian glands from intact, androgen- and/or placebo-treated rabbits were analyzed by histology, and glandular lipids were evaluated by gas chromatography, high-performance liquid chromatography (HPLC), and mass spectrometry. The rabbit tear film lipid layer was assessed by interferometry. RESULTS: In the current study androgen receptor protein existed within acinar epithelial cell nuclei of rat meibomian glands; androgen deficiency was associated with alterations in the lipid content of the rabbit meibomian gland; 19-nortestosterone treatment modulated the fatty acid profile in the total and neutral lipid fractions of the rabbit meibomian gland; and androgens did not appear to influence the gross morphology of meibomian tissue or to exert a demonstrable effect on the rabbit tear film lipid layer. CONCLUSIONS: The findings show that the meibomian gland is an androgen target organ and that androgens influence the lipid profile within this tissue. However, the extent to which androgens regulate the production of these lipids and whether this action may impact tear film stability remain to be determined. PMID- 11053271 TI - Plasminogen activator activity in tears after excimer laser photorefractive keratectomy. AB - PURPOSE: To quantify changes of plasminogen activator activity in tear fluid during corneal re-epithelialization after excimer laser photorefractive keratectomy (PRK). METHODS: Tear samples were collected with glass capillaries from 77 eyes of 42 patients immediately before and immediately after PRK treatment and on postoperative days 3 and 5. In 20 patients, the contralateral eye was similarly sampled to serve as control. Plasminogen activator activity in the tear samples was measured by a spectrophotometric method using human plasminogen and chromogenic peptide substrate, D-valyl-L-leucyl-L-lysine-p nitroanilide (S-2251). RESULTS: In tears of all eyes that underwent PRK, the plasminogen activator activities were lower immediately after PRK than were the preoperative values. For patient eyes with normal wound healing, tear plasminogen activator activities were significantly elevated above the preoperative level on the third postoperative day and then returned to the preoperative level by the fifth postoperative day. In contrast, tear plasminogen activator activities remained low through the third postoperative day in all (six) eyes in which haze developed after 3 to 6 months. The contralateral control eyes showed no appreciable change in plasminogen activator activity over the 5-day period. CONCLUSIONS: Plasminogen activator activity levels measured in tears of excimer laser PRK-treated eyes may serve as a predictor of wound healing. Extended low levels of plasminogen activator activity through the third postoperative day correlate with the development of corneal healing abnormalities (haze). The low plasminogen activator activity could be not only an accompanying sign but also a cause of defective corneal wound healing. PMID- 11053272 TI - Tear meniscus changes during cotton thread and Schirmer testing. AB - PURPOSE: To elucidate the effect of the cotton thread test (CT-T) and Schirmer test (S-T) on the tear reservoir by evaluating the radius of tear meniscus curvature. METHODS: The radii (R) of the central lower tear menisci were measured by a newly developed video meniscometer in 11 eyes of 11 normal volunteers (6 men, 5 women; mean age, 27.7 +/- 3.6 years [SD]) and 9 eyes of 9 patients with tear deficiency and severe dry eye in whom the puncta had been therapeutically occluded (9 women; mean age, 50.6 +/- 10.4 years). In this dry eye group, the absence of reflex tearing, coupled with the absence of lacrimal drainage due to punctal occlusion allowed more precise observation of the removal of tears from the meniscus. A 1-minute CT-T was performed, followed after an interval of 10 minutes by a 1-minute S-T. Tear meniscus curvature was documented before (R:(0)) and during the tests at 30 seconds (R(30)) and 60 seconds (R:(60)). RESULTS: In the normal group, respective R values (CT-T; S-T; mean +/- SD mm) were R(0) (0.26 +/- 0.11; 0.26 +/- 0. 07), R(30) (0.27 +/- 0.16; 0.20 +/- 0.13), and R(60) (0.29 +/- 0.15; 0.23 +/- 0.21); and in the dry eye group, respective R: values (CT-T; S T) were R(0) (0.59 +/- 0.23; 0.51 +/- 0.19), R(30) (0.52 +/- 0.25; 0.22 +/- 0.09), and R(60) (0.51 +/- 0.19; 0.21 +/- 0.08). It was demonstrated in the dry eye group that R was diminished more by the S-T than by the CT-T in the time course of the measurement (P = 0.01). In the dry eye group alteration of R occurred within the first 30 seconds, and in this group significant correlation was found between R(0) and the S-T result (r = 0.67; P = 0.05), and between R(60) R(0) and the S-T result (r = -0.81; P = 0.01). Also, there was a significant correlation between R(60)- R(0) and the S-T result in the normal group (r = 0.71; P = 0.02). There were no significant correlations between R(0) or R(60)- R(0) and the CT-T results in either group. CONCLUSIONS: These studies afford some insight into the dynamics of the Schirmer test, suggesting that wetting is influenced by the negative hydrostatic pressure within the tear meniscus. With the protocol used, no conclusion could be drawn about the relation between meniscus radius and wetting of the cotton thread. PMID- 11053273 TI - Temporal stimulation of corneal fibroblast wound healing activity by differentiating epithelium in vitro. AB - PURPOSE: To determine whether differentiating corneal epithelium can temporally stimulate fibroblast activity. METHODS: Corneal epithelial cells were cultured to confluence and then stimulated to mature into multilayered epithelia with addition of serum-containing medium. Differentiation was assessed morphologically and immunocytochemically using a monoclonal antibody to cytokeratin 3. At intervals after onset of differentiation serum-free conditioned medium was collected up to 28 days. Preliminary experiments deduced the optimum concentration of conditioned medium to be used for assessing fibroblast activity. Conditioned medium (25% vol/vol) was added to donor-matched corneal fibroblasts in migration chambers, WST-1 reagent proliferation assays, and fibroblast populated collagen gel contraction assays. Platelet-derived growth factor (PGDF) AB and interleukin (IL)-1beta in conditioned media were quantified by enzyme linked immunosorbent assay (ELISA). Fibroblast migration and collagen contraction assays were performed with concentrations of PDGF-AB. RESULTS: Conditioned medium collected from differentiating epithelium stimulated fibroblast migration and collagen gel contraction, with activity peaks occurring with medium collected on day 14 (P < 0.05). No significant difference was detected between fibroblast growth rates in each of the conditioned media. Levels of PDGF-AB increased during epithelial culture up to 22 days (up to approximately 360 pg/ml) with a subsequent decrease by 28 days. IL-1beta inversely correlated with fibroblast activity induced by conditioned medium, with a trough in concentration (2 pg/ml) occurring at 14 days. Both fibroblast migration and collagen contraction were stimulated in a dose-dependent manner by PDGF-AB. CONCLUSIONS: Corneal epithelium is capable of temporally stimulating fibroblast wound-healing characteristics during its differentiation. One of the growth factors potentially involved in this epithelial-stromal interaction is PDGF. This work demonstrated that developing epithelium (possibly similar to repairing epithelium in vivo) regulated fibroblast behavior and may indicate a mechanism of fibroblast recruitment to a wound after epithelial closure. PMID- 11053274 TI - Context-specific adaptation of pursuit initiation in humans. AB - PURPOSE: To determine if multiple states for the initiation of pursuit, as assessed by acceleration in the "open-loop" period, can be learned and gated by context. METHODS: Four normal subjects were studied. A modified step-ramp paradigm for horizontal pursuit was used to induce adaptation. In an increasing paradigm, target velocity doubled 230 msec after onset; in a decreasing paradigm, it was halved. In the first experiment, vertical eye position (+/-5 degrees ) was used as the context cue, and the training paradigm (increasing or decreasing) changed with vertical eye position. In the second experiment, with vertical position constant, when the target was red, training was decreasing, and when green, increasing. The average eye acceleration in the first 100 msec of tracking was the index of open-loop pursuit performance. RESULTS: With vertical position as the cue, pursuit adaptation differed between up and down gaze. In some cases, the direction of adaptation was in exact accord with the training stimuli. In others, acceleration increased or decreased for both up and down gaze but always in correct relative proportion to the training stimuli. In contrast, multiple adaptive states were not induced with color as the cue. CONCLUSIONS: Multiple values for the relationship between the average eye acceleration during the initiation of pursuit and target velocity could be learned and gated by context. Vertical position was an effective contextual cue but not target color, implying that useful contextual cues must be similar to those occurring naturally, for example, orbital position with eye muscle weakness. PMID- 11053275 TI - Cross-bridge kinetics of rabbit single extraocular and limb muscle fibers. AB - PURPOSE: To gain insights into the functional significance of myosin heavy-chain (MyHC) heterogeneity by comparing the mechanical kinetic properties of single rabbit extraocular muscle (EOM) fibers with those of limb fibers. EOMs are known to contain developmental and EOM-specific MyHCs in addition to those present in limb muscles, and MyHCs profoundly influence muscle mechanics. METHODS: Isometric cross-bridge kinetics were analyzed in Ca(2+)-activated single glycerinated fibers from rabbit EOM and limb fast and slow muscles at 15 degrees C by means of mechanical perturbation analysis. The plots of stiffness and phase against frequency display a characteristic frequency, f(min), at which stiffness is minimum, and phase shift is zero. The value of f(min) is independent of Ca(2+) or force level but reflects the kinetics of cross-bridge cycling. RESULTS: Analysis of 121 limb fast fibers gave f(min) values ranging from 10 to 26 Hz. f(min) for the 10 slow soleus fibers was 0.5 Hz. Analysis of 170 EOM fibers gave f(min) values in the range for fast limb fibers, but in addition yielded f(min) values below (4-9 Hz) and above (27-33 Hz) this range. CONCLUSIONS: The wider range of mechanical kinetic characteristics in EOM fibers compared with limb fibers is likely due to the expression of developmental (low f(min)) and EOM-specific (high f(min)) MyHCs in addition to isoforms present in adult limb muscles. The considerable diversity of functional characteristics in EOM fibers is likely to be important for rotating the eyeball at various speeds during tracking and for executing saccades over a wide range of angles. PMID- 11053276 TI - The degree of image degradation and the depth of amblyopia. AB - PURPOSE: To determine whether the depth of monocular form-deprivation amblyopia is dependent on the degree of retinal image degradation. METHODS: Chronic monocular form deprivation was produced in nine infant rhesus monkeys by securing one of three different strengths of diffuser spectacle lenses in front of the treated eye and a clear zero-powered lens in front of the fellow eye. Three infant monkeys reared with plano lenses in front of both eyes provided control data. The treatment lenses were worn continuously from approximately 3 weeks of age for periods ranging between 11 and 19 weeks. When the monkeys were approximately 18 months of age, psychophysical procedures were used to measure the effects of the rearing procedures on the spatial contrast sensitivity function for each eye. RESULTS: The treated eyes of all nine diffuser-reared monkeys showed contrast sensitivity deficits that were indicative of amblyopia. On average, the interocular grating acuity difference increased systematically from 0.6 octaves for the weakest diffuser lens to 2.3 +/- 0.7 and 3.5 +/- 0.8 octaves for the intermediate and strongest diffuser lenses, respectively. There was a close correspondence between the magnitude of the amblyopic deficits and the reduction in retinal image contrast produced by the diffuser lenses. CONCLUSIONS: The results demonstrate that the depth of monocular, nonstrabismic amblyopia is strongly influenced by the degree of retinal image degradation experienced early in life. PMID- 11053277 TI - Spatial localization in esotropia: does extraretinal eye position information change? AB - PURPOSE: To investigate the accuracy of spatial localization in children with a specific type of convergent strabismus, fully accommodative esotropia. METHODS: Two groups of children, with right and left fully accommodative esotropia, respectively, pointed at targets located centrally and eccentrically on a computer touchscreen without being able to see their hands. The size and the direction of the horizontal pointing responses were recorded under two conditions: when their eyes were aligned (wearing spectacles) and when they were squinting (not wearing spectacles). A group of children without strabismus but with hypermetropia were assessed as controls. RESULTS: For both fully accommodative groups, the pointing responses to the central target shifted in the direction of the nonsquinting eye when deviations were manifest. No difference was found for the eccentric targets. No difference was found for the hypermetropia group with any target. CONCLUSIONS: These results are consistent with an alteration in the extraretinal eye position information (efference copy, extraocular muscle proprioception, or both) that is used in spatial localization. PMID- 11053278 TI - Three-dimensional location of human rectus pulleys by path inflections in secondary gaze positions. AB - PURPOSE: Connective tissue pulleys serve as the functional mechanical origins of the extraocular muscles (EOMs). Anterior to these pulleys, EOM paths shift with gaze to follow the scleral insertions, whereas posterior EOM paths are stable in the orbit. Inflections in EOM paths produced by gaze shifts can be used to define the functional location of pulleys in three dimensions (3-D). METHODS: Contiguous magnetic resonance images in planes perpendicular to the orbital axis spanned the anteroposterior extents of 22 orbits of 11 normal adults with the eyes in central gaze, elevation, depression, abduction, and adduction. Mean EOM cross-sectional area centroids represented in a normalized, oculocentric coordinate system were plotted over the length of each EOM to determine paths. Path inflections were identified to define pulley locations in 3-D. RESULTS: All rectus EOM paths exhibited in secondary gaze positions distinct inflections 3 to 9 mm posterior to globe center, which were consistent across subjects. The globe center and the lateral rectus pulley translated systematically in the orbit with lateral gaze, whereas other pulleys remained stable relative to the orbit. CONCLUSIONS: Distinct inflections in rectus EOM paths in secondary gaze positions confirm the existence of pulleys and define their locations in 3-D. The globe and lateral rectus pulley translate systematically with gaze position. The EOM pulleys may simplify neural control of eye movements by implementing a commutative ocular motor plant in which commands for 3-D eye velocity are effectively independent of eye position. PMID- 11053279 TI - Visual control of postural orientation and equilibrium in congenital nystagmus. AB - PURPOSE: To investigate how humans with congenital nystagmus (CN) use visual information to stabilize and orient their bodies in space. METHODS: Center of foot pressure (COP) and head displacements in the lateral plane were recorded using a sway platform and Schottky barrier photodetector, respectively. In experiment 1, a comparison was made of the oscillatory characteristics of body sway with eyes open compared with eyes closed. Experiment 2 studied the postural readjustments made in response to absolute or relative motion (motion parallax) of objects in the visual scene, generated by lateral displacement of background scenery. RESULTS: Experiment 1 revealed that subjects with CN were not able to use visual information to stabilize COP but were able to stabilize the head at frequencies lower than 1 Hz. Experiment 2 showed that in response to the displacement of a visual display, for both absolute motion and motion parallax, subjects with CN reoriented their body in space in a manner similar to control subjects. CONCLUSIONS: The results suggest that despite involuntary eye movements, subjects with CN use orientation cues to control their posture, but not dynamic cues useful to control the rapid oscillations that are particularly important at the level of COP. These findings suggest that in CN, visual control of posture is restricted by low-frequency sampling of the visual scene. PMID- 11053280 TI - Waveform characteristics of manifest latent nystagmus. AB - PURPOSE: To examine the waveform characteristics of 37 subjects with manifest latent nystagmus (MLN) and determine the manner in which visual feedback influences the nature of the waveform. METHODS: Binocular recordings of the eye movements of all subjects were undertaken using an infrared tracking system. Subjects viewed the target binocularly and monocularly in primary gaze. The effect of visual feedback on the nature of the MLN waveform was examined by either removing the fixation target or by progressively stabilizing the target in relation to the retina. This progressive stabilization was achieved by feeding back the eye movement signal to move an otherwise stationary target. RESULTS: Four types of MLN were distinguished on the basis of the fixation characteristics seen during binocular and monocular viewing. First, under binocular viewing conditions, subjects could theoretically exhibit stable fixation (type 1 MLN). In addition, three other MLN types were recorded during binocular fixation: conjugate horizontal square-wave jerks (type 2 MLN), conjugate torsional nystagmus (type 3 MLN) and conjugate horizontal jerk MLN waveforms (type 4 MLN). Monocular viewing always gave rise to a conjugate horizontal jerk MLN waveform for each of the four types of MLN. More than 80% of the subjects exhibited either type 3 or type 4 MLN, both of which conform with previous classic descriptions of MLN. Much less common was type 2 MLN. Type 1 MLN (conventionally referred to as a latent nystagmus) appeared to be a rare occurrence. In addition to the two classic linear and decelerating MLN slow phases, four additional slow-phase shapes with either saccadic or pendular elements were recorded and described. Removing visual feedback generally reduced the mean slow-phase velocity and the number of fast phases. For each subject some variability of the slow-phase class was documented from session to session. CONCLUSIONS: Four types of MLN have been described. Their differences are based on their binocular oculomotor behavior, and it is proposed that type 1 MLN and type 4 MLN represent the absolute states and types 2 and 3 the intermediate levels of the MLN spectrum. All types of MLN appear to be strongly visually driven and are largely dependent on the attentional state of the subject and the target conditions. Six different classes of slow phase were found among the four MLN types. The introduction of visual feedback had an immediate effect on the subsequent slow phase or fast phase. It is likely that adaptation mechanisms are in play after a period of visual feedback. PMID- 11053281 TI - A spatial frequency-doubling illusion-based pattern electroretinogram for glaucoma. AB - PURPOSE: A pattern electroretinogram (PERG) in which stimuli displaying the frequency-doubling (FD) illusion are presented simultaneously to multiple parts of the visual field was evaluated for its ability to diagnose glaucoma. This multiregion FD PERG is referred to in the current study as the MFP. METHODS: The nine stimulus regions were temporally modulated at incommensurate frequencies typically producing an FD percept. Two other spatial scales of the stimuli were also investigated. The sensitivity and specificity of MFP were examined using linear and quadratic discriminant methods. RESULTS: Even with the simpler linear discriminant classification, sensitivities and specificities of 100% were obtained in eyes with moderate to severe glaucoma. Of eyes with glaucoma strongly suspected, 67% were classified as being glaucomatous. Stimulus patterns having differing spatial scales produced different PERG visual field dependencies. CONCLUSIONS: The differing results for the 16-fold change in spatial scale may reflect the accessing of different mechanisms. The MFP method appears to have significant value for the diagnosis of glaucoma. PMID- 11053282 TI - Comparing a parallel PERG, automated perimetry, and frequency-doubling thresholds. AB - PURPOSE: A pattern electroretinogram (PERG) simultaneously displaying the frequency-doubling (FD) illusion in nine parts of the visual field was compared with two other methods for ability to detect glaucoma. This multiregion FD PERG (MFP) was compared with results from achromatic automated perimetry and psychophysical tests using FD stimuli. METHODS: MFP data were compared with that from the Humphrey Field Analyser (HFA; Humphrey, San Leandro, CA) 24-2 program. Contrast thresholds were also determined in different visual field locations for FD stimuli. Thin-plate spline methods were used to derive comparisons from the tests, each of which sampled the visual field differently. RESULTS: Significant correlation with HFA could be obtained, providing seven to nine (of nine) MFP amplitudes were themselves significant. Evidence showed that both the psychophysical tests using FD stimuli and the MFP detect glaucomatous damage not detected by the HFA. CONCLUSIONS: The comparisons between HFA perimetry, the MFP, and FD thresholds indicate that both FD-based tests quantify a form of diffuse loss in early glaucoma as well as the scotomas of later glaucoma. PMID- 11053283 TI - Human trabecular meshwork cells secrete neurotrophins and express neurotrophin receptors (Trk). AB - PURPOSE: The purpose of this study was to compare the mRNA expression of neurotrophins (NTs) and NT receptors (Trk) in cultured human trabecular meshwork (HTM) cells and ex vivo HTM tissues, to immunolocalize both NT and Trk receptors in cultured HTM cells, and to demonstrate secretion of NTs by HTM cells. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of NT and Trk receptor mRNAs in early-passaged, cultured HTM cells from donors of several ages. RT-PCR was used on ex vivo HTM tissues from donors to compare and contrast mRNA expression with cell culture results. In addition, immunohistochemistry was used to localize the translated NT and low- (p75) and high- (Trk) affinity NT receptor proteins within cultured HTM cells and trabecular meshwork tissues. Last, enzyme-linked immunoassay (ELISA) was used to demonstrate secretion of NTs by HTM cells. RESULTS: Amplification products of the expected size for NTs were detected in both cultured HTM cells and ex vivo HTM tissues. Specifically identified were amplification products for the following NTs: NGF, BDNF, NT-3, and NT-4. Amplification products for the full-length Trk A and Trk C high-affinity receptor were observed, as well as truncated isoforms for Trk B and Trk C. No amplification products were produced for the full-length Trk B receptor nor for the low-affinity p75 receptor. Immunohistochemistry indicated that proteins for the various NTs and full-length and truncated Trk receptors were translated by cultured HTM cells and tissues. Immunoassays (ELISA) detected BDNF, NT-4, NGF, and NT-3 in the culture media from HTM cells. CONCLUSIONS: The results demonstrate, for the first time, mRNA expression for NT and Trk receptors by both cultured HTM cells and ex vivo HTM tissues. NTs were immunolocalized in HTM tissues and cultured HTM cells are capable of secreting NTs. Specific NTs acting through high-affinity Trk receptors within the HTM may be involved in maintaining the normal function of this complex tissue. PMID- 11053284 TI - Localization of myocilin to the golgi apparatus in Schlemm's canal cells. AB - PURPOSE: Biochemical and genetic evidence suggests that overexpression of or mutations in myocilin within the cells of the aqueous humor outflow pathway play a significant role in the development of steroid-induced and several other open angle glaucomas. As a baseline to understanding the normal and pathologic function of myocilin, we determined the subcellular localization of myocilin in steroid-treated human Schlemm's canal endothelial (SC) cells. METHODS: SC cells were grown to confluence, treated with dexamethasone for 10 days, and then stained using antibodies against myocilin, tubulin, or beta-COP (a specific golgi protein) or vital stains for endoplasmic reticulum (ER) and golgi. Brefeldin A (BFA) and nocodazol (NZ) were used to disrupt the golgi or microtubules. RESULTS: The authors found that myocilin staining was (a) always centered around the centrosome, (b) very similar to the pattern seen with NBD-ceramide, (c) was disrupted in characteristic ways by BFA and NZ and (d) showed extensive colocalization with beta-COP. CONCLUSIONS: Results indicate that myocilin is localized to the golgi in SC cells. Such localization is consistent with myocilin being processed for secretion but is also consistent with sequence analysis and other data that suggest that myocilin or myocilin mutations might be targeted to the cytoplasmic face of the golgi, and under some circumstances play a role in or interfere with golgi or vesicle function. How such interference could eventually lead to open angle glaucoma is discussed. PMID- 11053285 TI - Prevention of acute allergic conjunctivitis and late-phase inflammation with immunostimulatory DNA sequences. AB - PURPOSE: To evaluate the therapeutic potential of immunostimulatory sequence oligodeoxynucleotides (ISS-ODN) administration in ocular allergy, using a mouse model of ragweed-specific conjunctivitis. METHODS: A murine model of allergic conjunctivitis involving SWR/J mice sensitized and challenged with short ragweed was used to test immunostimulatory DNA sequences for therapeutic potential. ISS ODN or control ODN (0.1 mg/mouse) was administered intraperitoneally or topically to the conjunctiva 3 days before final allergen challenge. Multiple parameters of clinical symptoms evident during the acute-phase reaction and the cellular components of the late-phase reaction were evaluated in both groups of mice. RESULTS: All parameters of clinical symptoms were markedly inhibited after intraperitoneal injection of ISS-ODN, whereas topical application to the conjunctiva did not inhibit clinical symptoms significantly. Remarkably, a single topical treatment with ISS-ODN (as well as by intraperitoneal injection) completely inhibited both eosinophilia and neutrophilia in the late-phase reaction. CONCLUSIONS: Systemic or conjunctival administration of ISS-ODN was shown to significantly inhibit allergic responses in this mouse model. This indicates that ISS-ODN may be an effective form of immunotherapy for this class of allergic disease. PMID- 11053286 TI - Impaired eosinophil recruitment to the cornea in P-selectin-deficient mice in Onchocerca volvulus keratitis (River blindness). AB - PURPOSE: A murine model of helminth-induced keratitis (river blindness) that is characterized by a biphasic recruitment of neutrophils (days 1-3) and eosinophils (days 3+) to the cornea has been developed. The purpose of this study was to determine the relative contribution of P- and E-selectin in recruitment of these inflammatory cells from limbal vessels to the corneal stroma. METHODS: P- and E selectin gene knockout (-/-) mice were immunized with antigens extracted from the parasitic helminth Onchocerca volvulus. One week after the last immunization, parasite antigens were injected directly into the corneal stroma. Mice were killed on days 1 and 3 postchallenge, and eyes were immunostained with either anti-eosinophil major basic protein (MBP) or with anti-neutrophil Ab. The number of cells in the cornea was determined by direct counting. RESULTS: Recruitment of eosinophils to the cornea was significantly impaired in P-selectin(-/-) mice (63.9% fewer eosinophils on day 1 [P: = 0.0015], and 61% fewer on day 3 [P: < 0.0001]) compared with control C57BL/6 mice. In contrast, P-selectin deficiency had no effect on neutrophil recruitment to the cornea. There was no inhibition of eosinophil and neutrophil migration to the corneas of E-selectin(-/-) mice, indicating that there is no direct role for this adhesion molecule in helminth induced keratitis. CONCLUSIONS: The present study demonstrates that P-selectin is an important mediator of eosinophil recruitment to the cornea. P-selectin interactions may therefore be potential targets for immunotherapy in eosinophil mediated ocular inflammation. PMID- 11053287 TI - Participation of pigment epithelium of iris and ciliary body in ocular immune privilege. 2. Generation of TGF-beta-producing regulatory T cells. AB - PURPOSE: To determine whether T cells exposed to cultured iris and ciliary body pigment epithelial (I/CB PE) cells acquire the capacity to modify the activation, differentiation, and effector functions of bystander T cells, and if so, to identify the mechanism. METHODS: T cells from naive BALB/c mice were cultured with I/CB PE cells, x-irradiated, and used as regulators (a) of T-cell activation in vitro and (b) of delayed hypersensitivity expression in vivo. Neutralizing anti-TGF-beta and -IL-10 antibodies were used to abolish regulatory function. T cell activation was assessed for proliferation by [(3)H]thymidine incorporation and for IL-2, IFN-gamma, IL-4, and IL-10 production by semi-quantitative RT-PCR for mRNA and by supernatant analysis by ELISA. I/CB PE-exposed T cells were evaluated for mRNA content of IFN-gamma, IL-4, TNF-alpha, TGF-beta1, TGF-beta2, and IL-10, and their supernatants were analyzed for content of TGF-beta. RESULTS: T cells exposed to I/CB PE cells inhibited anti-CD3-driven activation of bystander naive T cells in vitro and suppressed the expression of delayed hypersensitivity in vivo. Bystander T cells cocultured with I/CB PE-exposed T cells failed to proliferate and secreted high levels of IL-4 and IL-10 but low amounts of IL-2 and IFN-gamma. Regulation of bystander T-cell activation was mediated via enhanced secretion of TGF-beta by I/CB PE-exposed T cells. CONCLUSIONS: T cells exposed to cultured I/CB PE cells were induced to secrete active and latent TGF-beta, which conferred on the T cells the capacity to inhibit the differentiation as well as the effector function of Th1-type cells. PMID- 11053288 TI - Substance P differentially stimulates IL-8 synthesis in human corneal epithelial cells. AB - PURPOSE: To determine whether substance P (SP), a neuropeptide with proinflammatory properties, specifically interacts with human corneal epithelial cells to stimulate synthesis of the chemokines interleukin (IL)-8, monocyte chemo attractant protein (MCP)-1, and regulated on activation normal T-cell expressed and secreted (RANTES) protein. METHODS: Primary cultures of human corneal epithelial cells were established from human corneas. Expression of the SP receptor neurokinin (NK)-1 was determined by both the reverse transcription polymerase chain reaction (RT-PCR) and radiolabeled saturation binding experiments. Synthesis of chemokine-specific RNA in cells stimulated with SP was analyzed by RT-PCR, and quantitation of chemokine protein synthesis was achieved by enzyme-linked immunosorbent assay. RESULTS: Human corneal epithelial cells expressed NK-1 mRNA and bound SP with a K:(d) characteristic of NK-1. Exposure of cells to SP had no effect on IL-8-specific mRNA synthesis, whereas it increased the half-life of IL-8 transcripts by more than twofold, resulting in significant enhancement of IL-8 synthesis. The capacity of SP to bind to corneal epithelial cells and to induce IL-8 synthesis was abrogated in the presence of a specific NK 1 receptor antagonist. In contrast to IL-8, exposure of cells to SP did not stimulate synthesis of MCP-1 or RANTES. CONCLUSIONS: The results suggest that human corneal cells express NK-1 receptors that specifically bind SP and induce IL-8 synthesis by stabilizing the chemokine's transcripts. PMID- 11053289 TI - MRI of the human eye using magnetization transfer contrast enhancement. AB - PURPOSE: To determine the feasibility of using magnetization transfer contrast enhanced magnetic resonance imaging (MRI) to track cataractous lens changes. METHODS: A fast spin-echo sequence was modified to include a magnetization transfer contrast (MTC) preparation pulse train. This consisted of twenty 8.5 msec sinc pulses, 1200 Hz upfield from the water resonance and 1.2-Hz power. The MTC preparation pulse was followed by acquisition through fast spin-echo imaging. The imaging parameters were number of excitations (NEX) = 1, echo time (TE) = 14 msec, recovery time (TR) = 2 sec, echo train length of eight echos, and a matrix size of 256 x 160. To reduce motion artifacts, the volunteers were asked to fixate on a blinking LED. Normal and MTC-enhanced images were acquired from normal volunteers and volunteers with nuclear or cortical cataracts. RESULTS: The eye was adequately imaged, with few motion artifacts appearing. The lens was well resolved, despite the short T(2). The cornea and ciliary body were also clearly visible. In the lens, resolution of the epithelium and cortex were enhanced with MTC. In addition, contrast-to-noise ratios were measured for each image. Examination of the contrast-to-noise ratio confirmed that MTC increased the contrast between the nucleus and cortex. Unenhanced MRIs showed significant differences between the cortex of normal volunteers and volunteers with cataracts. MTC-enhanced images improved the sensitivity to changes in the nucleus. CONCLUSIONS: In this preliminary study, we were able to use MTC-enhanced MRI to obtain high-contrast images of the human lens. Regular and enhanced MRIs detected statistically significant differences between normal and cataractous lenses. PMID- 11053290 TI - Quantification of posterior capsular opacification in digital images after cataract surgery. AB - PURPOSE: To describe a software program developed to provide an objective assessment of the amount of posterior capsular opacification (PCO) in high resolution digital images of the posterior capsule after cataract surgery. METHODS: Images are analyzed by a set protocol of defining the area of the posterior capsule, removing the Purkinje light reflexes by intensity segmentation, contrast enhancement, filtering to enhance low-density PCO, and variance analysis using a co-occurrence matrix to assess texture. The accuracy of the system was tested for validity and repeatability. RESULTS: The software developed has been demonstrated to be an objective method of quantifying PCO. In validation tests, the image analysis-derived measure of PCO showed good agreement with clinically derived measures of PCO. Clinicians assessed PCO on a computer screen image and also under slit lamp examination (Pearson correlation coefficient for both methods >0.92). The entire acquisition and analysis system was demonstrated to have a confidence limit for 2 SDs of 9.8% for group data. CONCLUSIONS: This system is capable of producing an accurate and reproducible measure of PCO that is relevant to assessing techniques of PCO prevention. PMID- 11053291 TI - In vitro filament-like formation upon interaction between lens alpha-crystallin and betaL-crystallin promoted by stress. AB - PURPOSE: To determine whether alpha-crystallin is capable of forming filament like structures with other members of the crystallin family. METHODS: Water soluble crystallins were isolated from calf lenses and fractionated into alpha-, betaH-, betaL-, and gamma-crystallins according to standard procedures. Chaperone like activity of alpha-crystallin was determined in control and UV-A-irradiated lenses by the heat-induced aggregation assay of betaL-crystallin. Protein samples from this assay were analyzed by electron microscopy. In vitro filament formation was examined by transmission immunoelectron microscopy using specific antibodies directed against the crystallins. Involvement of intermediate filament constituents was excluded by the results of Western blot analysis, which were all negative. Moreover, the in vitro amyloid fibril interaction test using thioflavin T (ThT) was also performed. RESULTS: At the supramolecular level heating at 60 degrees C has no effect on the morphologic appearance of alpha-crystallin as observed by transmission electron microscopy. Moreover alpha-crystallin obtained from UV-A-irradiated lenses shows a virtually identical shape. However, heating in the presence of betaL-crystallin results in the formation of filament-like alphabeta-hybrids as demonstrated by immunoelectron microscopy using specific antibodies directed either against alpha- or betaL-crystallin. Parallel experiments with alpha-crystallin derived from UV-A-irradiated lenses showed even more pronounced filamentous structures, compared with the controls. Nonetheless, we were able to show that the UV-light treatment affected the chaperone-like capacity of alpha-crystallin, as revealed by a diminished ability to inhibit in vitro denaturation of betaL-crystallin. To exclude the presence of cytoskeletal contamination in the crystallin preparations, vimentin antibodies were also tested. These latter experiments were negative. The filamentous nature of the hybrids was further confirmed by the results obtained with the ThT assay earlier applied for the detection of amyloid fibrils. CONCLUSIONS: Crystallin hybrids have previously been detected in the water-soluble lens crystallin fraction. Our findings indicate that such endogenous hybrids, formerly called "rods," may result from stress-induced interaction between alpha-crystallin and other lens constituents such as betaL-crystallin. Because the hybrid formation is enhanced when alpha-crystallin from UV-A-irradiated lenses is used as one of the two components of the hybrid, one can only speculate that this formation may be one of the factors leading to UV-A cataract. PMID- 11053292 TI - Growth and differentiation of human lens epithelial cells in vitro on matrix. AB - PURPOSE: To characterize the growth and maturation of nonimmortalized human lens epithelial (HLE) cells grown in vitro. METHODS: HLE cells, established from 18 week prenatal lenses, were maintained on bovine corneal endothelial (BCE) extracellular matrix (ECM) in medium supplemented with basic fibroblast growth factor (FGF-2). The identity, growth, and differentiation of the cultures were characterized by karyotyping, cell morphology, and growth kinetics studies, reverse transcription-polymerase chain reaction (RT-PCR), immunofluorescence, and Western blot analysis. RESULTS: HLE cells had a male, human diploid (2N = 46) karyotype. The population-doubling time of exponentially growing cells was 24 hours. After 15 days in culture, cell morphology changed, and lentoid formation was evident. Reverse transcription-polymerase chain reaction (RT-PCR) indicated expression of alphaA- and betaB2-crystallin, fibroblast growth factor receptor 1 (FGFR1), and major intrinsic protein (MIP26) in exponential growth. Western analyses of protein extracts show positive expression of three immunologically distinct classes of crystallin proteins (alphaA-, alphaB-, and betaB2-crystallin) with time in culture. By Western blot analysis, expression of p57(KIP2), a known marker of terminally differentiated fiber cells, was detectable in exponential cultures, and levels increased after confluence. MIP26 and gamma-crystallin protein expression was detected in confluent cultures, by using immunofluorescence, but not in exponentially growing cells. CONCLUSIONS: HLE cells can be maintained for up to 4 months on ECM derived from BCE cells in medium containing FGF-2. With time in culture, the cells demonstrate morphologic characteristics of, and express protein markers for, lens fiber cell differentiation. This in vitro model will be useful for investigations of radiation-induced cataractogenesis and other studies of lens toxicity. PMID- 11053293 TI - Dystrobrevin localization in photoreceptor axon terminals and at blood-ocular barrier sites. AB - PURPOSE: Dystrobrevin is a newly discovered dystrophin-associated protein with multiple sites for phosphorylation on tyrosine residues. In the present study, the cellular distribution and subcellular localization of dystrobrevin were examined in the adult rat retina, cornea, lens, iris, ciliary body, and cultured Muller cells. METHODS: Immunoblot analysis, confocal laser scanning microscopy, and immunoelectron microscopy were used to examine dystrobrevin expression. RESULTS: Immunoblot analysis showed that an approximately 87-kDa band was expressed predominantly in the lens, retina, iris and ciliary body, whereas an approximately 60-kDa band was expressed in cultured Muller cells, cornea, retina, iris, and ciliary body. Confocal microscopy demonstrated dystrobrevin in the inner limiting membrane, outer plexiform layer, and retinal pigment epithelium and around blood vessels in the retina. At the ultrastructural level, dystrobrevin was localized under cell membranes of rod spherules and cone pedicles of photoreceptor cell terminals but often was found in the cytoplasm of endothelial cells and Muller cells. Furthermore, dystrobrevin was colocalized with beta-dystroglycan in corneal endothelium; lens, iris, and ciliary epithelia; and cultured Muller cells. CONCLUSIONS: The present study demonstrates that dystrobrevin is expressed in neurons, glia, and endothelial cells in the rat retina. In addition, dystrobrevin is localized at the blood-ocular barrier sites in extraocular tissue. These data suggest that dystrobrevin plays an important role in visual function. PMID- 11053294 TI - EIU in the rat promotes the potential of syngeneic retinal cells injected into the vitreous cavity to induce PVR. AB - PURPOSE: To determine whether syngeneic retinal cells injected in the vitreous cavity of the rat are able to initiate a proliferative process and whether the ocular inflammation induced in rats by lipopolysaccharide (LPS) promotes this proliferative vitreoretinopathy (PVR). METHODS: Primary cultured differentiated retinal Muller glial (RMG) and retinal pigmented epithelial (RPE) cells isolated from 8 to 12 postnatal Lewis rats were injected into the vitreous cavity of 8- to 10-week-old Lewis rats (10(5) cells/eye in 2 microlieter sterile saline), with or without the systemic injection of 150 microgram LPS to cause endotoxin-induced uveitis (EIU). Control groups received an intravitreal injection of 2 microliter saline. At 5, 15, and 28 days after cell injections, PVR was clinically quantified, and immunohistochemistry for OX42, ED1, vimentin (VIM), glial fibrillary acidic protein (GFAP), and cytokeratin was performed. RESULTS: The injection of RMG cells, alone or in combination with RPE cells, induced the preretinal proliferation of a GFAP-positive tissue, that was enhanced by the systemic injection of LPS. Indeed, when EIU was induced at the time of RMG cell injection into the vitreous cavity, the proliferation led to retinal folds and localized tractional detachments. In contrast, PVR enhanced the infiltration of inflammatory cells in the anterior segment of the eye. CONCLUSIONS: In the rat, syngeneic retinal cells of glial origin induce PVR that is enhanced by the coinduction of EIU. In return, vitreoretinal glial proliferation enhanced the intensity and duration of EIU. PMID- 11053295 TI - A high association with cone dystrophy in Fundus albipunctatus caused by mutations of the RDH5 gene. AB - PURPOSE: To analyze the RDH5 gene in patients with fundus albipunctatus with and without cone dystrophy and to determine whether the disease is stationary or progressive and whether the cone dystrophy is a part of fundus albipunctatus or a separate disease. METHODS: Fourteen patients from 12 separate Japanese families with fundus albipunctatus were examined. Six of the patients from 6 families also had a cone dystrophy. Genomic DNA was extracted from leukocytes of the peripheral blood, and exons 2, 3, 4, and 5 of the RDH5 gene were amplified by polymerase chain reaction and were directly sequenced. A complete ophthalmic examination was performed including best-corrected visual acuity, slit-lamp examination, indirect ophthalmoscopy, fundus photography, and electroretinography. RESULTS: In all the patients, either a homozygous mutation or compound heterozygous mutations in the RDH5 gene were identified. The identified mutations were nucleotide position (nt) 103 G to A (Gly35Ser), nt 319 G to C (Gly107Arg), nt 394 G to A (Val132Met), nt 719 G insertion (frame shift), nt 839 G to A (Arg280His), nt 841 T to C (Tyr281His), and nt 928 C to GAAG (Leu310 to GluVal). All these mutations except the Arg280His were new. The nt 928 C to GAAG mutation was detected in patients with and without cone dystrophy. Cone dystrophy was most frequently seen in patients over 40 years old. CONCLUSIONS: Fundus albipunctatus either with or without cone dystrophy is caused by mutations of the RDH5 gene. Cone dystrophy is frequently observed in elderly patients with fundus albipunctatus. The conclusion was reached that the mutations of the RDH5 gene caused a progressive cone dystrophy as well as night blindness. PMID- 11053296 TI - Mutations in the 11-cis retinol dehydrogenase gene in Japanese patients with Fundus albipunctatus. AB - PURPOSE: To detect mutations in the RDH5 gene encoding 11-cis retinol dehydrogenase in patients from Japan with fundus albipunctatus. METHODS: Polymerase chain reaction and direct genomic sequencing techniques were used to detect mutations of the RDH5 coding exons (exons 2-5) in two unrelated patients with fundus albipunctatus. Selected alleles that altered the coding region or intron splice sites were evaluated further through segregation analysis in the families of the index cases. RESULTS: Two novel RDH5 mutations were identified. One of these was a missense mutation Val264Gly in exon 5, and the other was an in frame insertion of 3 bp in exon 5. CONCLUSIONS: The data indicate that mutations in RDH5 are the primary cause of fundus albipunctatus. PMID- 11053297 TI - Age-related retinal pigment epithelium and Bruch's membrane degeneration in senescence-accelerated mouse. AB - PURPOSE: To investigate age-related changes in the retinal pigment epithelium (RPE), Bruch's membrane, and choriocapillaris in the senescence-accelerated mouse (SAM). METHODS: The external and eyecup features and light and electron microscopic findings were examined in three male and two female mice of a senescence-prone mouse strain (SAM P(8)) monthly for 12 months. These results were compared with those in age-matched mice of similar background but senescence resistant (SAM R(1)). Choroidal vascular casts were prepared at 12 months in seven mice each of the SAM P(8) and SAM R(1) strains. Quantitative analysis of area of choriocapillaris was performed by automated image analysis, and the results were analyzed by paired Student's t-test. RESULTS: We found in the SAM P(8) strain that hair loss, coarseness of hair texture, and ulceration of skin appeared and increased as the age advanced (at approximately 5-9 months). Eyecup examination showed no differences. Light and electron microscopy revealed progressively more prominent abnormalities in the RPE and Bruch's membrane mice older than 10 months. Two of the five SAM P(8) mice older than 11 months showed what appeared to be intra-Bruch's membrane choroidal neovascularization. The RPE and Bruch's membrane appeared normal in the SAM R(1) strain. In the SAM P(8), vascular casts of the choriocapillaris showed a mild but significant decrease in vascular area when compared with the SAM R(1) strain at 12 months (P = 0.011). CONCLUSIONS: Senescence accelerated mice develop progressive age-related changes in the RPE-Bruch's-choriocapillaris complex that have features that may be relevant in the study of age-related macular changes in humans. PMID- 11053298 TI - Cataractogenic lens injury prevents traumatic ganglion cell death and promotes axonal regeneration both in vivo and in culture. AB - PURPOSE: To examine and quantify neuroprotective and neurite-promoting activity on retinal ganglion cells (RGCs) after injury of the lens. METHODS: In adult albino rats, penetrating lens injury was performed by intraocular injection. To test for injury-induced neuroprotective effects in vivo, fluorescence-prelabeled RGCs were axotomized by subsequent crush of the optic nerve (ON) with concomitant lens injury to cause cataract. The numbers of surviving RGCs were determined in retinal wholemounts and compared between the different experimental and control groups. To examine axonal regeneration in vivo, the ON was cut and replaced with an autologous piece of sciatic nerve (SN). Retinal ganglion cells with axons that had regenerated within the SN under lens injury or control conditions were retrogradely labeled with a fluorescent dye and counted on retinal wholemounts. Neurite regeneration was also studied in adult retinal explants obtained either after lens injury or without injury. The numbers of axons were determined after 1 and 2 days in culture. Putative neurotrophins (NTs) were studied within immunohistochemistry and Western blot analysis. RESULTS: Cataractogenic lens injury performed at the same time as ON crush resulted in highly significant rescue of 746 +/- 126 RGCs/mm(2) (mean +/- SD; approximately 39% of total RGCs) 14 days after injury compared with controls without injury or with injection of buffer into the vitreous body (30 +/- 18 RGCs/mm(2)). When lens injury was performed with a delay of 3 days after ON crush, 49% of RGCs survived, whereas delay of 5 days still rescued 45% of all RGCs. In the grafting paradigm virtually all surviving RGCs after lens injury appeared to have regenerated an axon within the SN graft (763 +/- 114 RGCs/mm(2) versus 79 +/- 17 RGCs/mm(2) in controls). This rate of regeneration corresponds to approximately 40% of all RGCs. In the regeneration paradigm in vitro preceding lens injury and ON crush 5 days previous resulted in a maximum of regeneration of 273 +/- 39 fibers/explant after 1 day and 574 +/- 38 fibers/explant after 2 days in vitro. In comparison, in control retinal pieces without lens injury 28 +/- 13 fibers/explant grew out at 1 day, and 97 +/- 37 fibers/explant grew out at 2 days in culture. Immunohistochemical and Western blot analysis of potential NTs in the injured lens revealed no expression of ciliary neurotrophic factor (CNTF), brain-derived neurotrophic factor (BDNF), NT-4, nerve growth factor (NGF), and basic fibroblast growth factor (bFGF). CONCLUSIONS: The findings indicate that the lens contains high neuroprotective and neuritogenic activity, which is not caused by NT. Compared with the data available in the literature, this neuroprotection is quantitatively among the highest ever reported within the adult rat visual system. PMID- 11053299 TI - Endothelin-3 regulation of retinal hemodynamics in nondiabetic and diabetic rats. AB - PURPOSE: To investigate the mechanisms of action of endothelin (ET)-3 on the regulation of retinal hemodynamics in diabetic and nondiabetic rats. METHODS: Retinal blood flow changes were measured using video fluorescein angiography. Measurements were made before and after intravitreal injections of different ET-3 concentrations in nondiabetic rats and rats with streptozotocin (STZ)-induced diabetes. The effect of ET-3 on retinal blood flow was also investigated in nondiabetic rats after pretreatment with N:(G)-monomethyl-L-arginine (L-NMMA), a nitric oxide synthase (NOS) inhibitor; BQ-788, an ET receptor B (ETB) antagonist; and BQ-123, an ET receptor A (ETA) antagonist. Control animals were injected intravitreally with vehicle alone. RESULTS: In nondiabetic rats, ET-3 induced a dose-dependent rapid increase in retinal blood flow 2 minutes after intravitreal injection (maximal at 10(-)(8) M, P < 0. 01) followed 15 and 30 minutes after ET 3 injection by dose-dependent decreases in retinal blood flow (maximal effect at 10(-)(6) M, P < 0.05). The ET-3-stimulated retinal blood flow increase was inhibited by 10(-)(4) M BQ-788 (P < 0.01) and 10(-)(3) M L-NMMA (P < 0.05). The ET-3-stimulated decrease in retinal blood flow at later times (15 minutes) was inhibited (P < 0.03) by 10(-4) M BQ-123. In diabetic rats, baseline retinal blood flows were decreased compared with nondiabetic rats (P < 0.01), showed dose dependent increases 2 minutes after ET-3 injection (P < 0.03), and at later times remained significantly increased (P < 0.05) in contrast to flows in nondiabetic rats. CONCLUSIONS: The ET-3-induced initial rapid retinal blood flow increase in nondiabetic rats is mediated by the ET-3/ETB and NOS action. The subsequent retinal blood flow decrease is mediated by ET-3/ETA action. Diabetic rats showed comparable ET-3-induced retinal blood flow increases indicating normal ET-3/ETB action. However, at later times, retinal blood flow remained increased, suggesting an abnormal ET-3/ETA action. PMID- 11053300 TI - Photodynamic therapy using Lu-Tex induces apoptosis in vitro, and its effect is potentiated by angiostatin in retinal capillary endothelial cells. AB - PURPOSE: To examine the effect of combining angiostatin with photodynamic therapy (PDT) using Lutetium Texaphyrin (Lu-Tex; Alcon, Fort Worth, TX) as a photosensitizer in bovine retinal capillary endothelial (BRCE) and retinal pigment epithelial (RPE) cells and to determine the mode of PDT-induced cell death in these cell lines. METHODS: Cultured BRCE and RPE cells were incubated with angiostatin (500 ng/ml) for 18 hours and subjected to Lu-Tex/PDT, using treatment parameters previously optimized (3 microgram/ml Lu-Tex for 30 minutes followed by timed irradiation at 732 nm). Cellular survival was assessed after a 1-week cellular proliferation. Data were analyzed using Student's t-test. Caspase 3 activity was monitored in cells after PDT using a fluorogenic substrate, (Asp Glu-Val-Asp)-AFC (7-amino-4-trifluoromethyl coumarin) [DEVD-AFC], of caspase 3. After PDT, expression of Bcl-2, Bcl-x(L), Bax, and Bak was also examined in cell lysates by Western blot analysis. RESULTS: A synergistic cytotoxic effect of angiostatin and Lu-Tex/PDT was observed in BRCE cells at all fluences used (5, 10, and 20 J/cm(2); P 0.95) for 64 h. In contrast to controls, enterally fed hyperoxic animals did not lose weight and had smaller pleural effusions and wet to-dry weight ratios (a measure of lung edema) that were not different from room air controls. Enterally fed rats exposed to hyperoxia had increased levels of mRNA for the Na(+)-K(+)-ATPase alpha(1)- and beta(1)-subunits and glutathione peroxidase. These findings suggest that maintenance of nutrition during an oxidative lung injury reduces lung edema, perhaps by allowing for continued expression and function of protective proteins such as the Na(+)-K(+)-ATPase. PMID- 11053324 TI - Simplified rat intubation using a new oropharyngeal intubation wedge. AB - Our new oropharyngeal intubation wedge made from a plastic 3-ml syringe has been used successfully for the expansion of the oropharyngeal cavity and visualization of vocal cords for endotracheal intubation in the rat. All the animals we used tolerated the intubation and ventilation procedures in a series of experiments. After the proper setting of the respirator, vital signs were maintained within normal range. The postmortem examination and measurements in the upper airway confirmed that the endotracheal tube was properly sited and also demonstrated the precise size of the device that should be used. The main advantages of this method include low cost, simplicity, and reliability. Furthermore, because no expensive, elaborate, difficult-to-operate, or hard-to-get special equipment is needed, this technique can be used in every laboratory. PMID- 11053325 TI - Initial ventilatory and circulatory responses to dynamic exercise are slowed in the elderly. AB - To elucidate the characteristics of ventilatory and circulatory responses at the onset of brief and light exercise in the elderly, 13 healthy, elderly men, aged 66.8 yr (mean), exerted bilateral leg extension-flexion movements for only 20 s with a weight around each ankle, with each weight being approximately 2.5% of their body mass. Similar movements were passively performed on the subjects by the experimenters. These results were compared with those of 13 healthy, young men (22.9 yr). Minute ventilation increased at the onset of voluntary exercise and passive movements in both groups but showed a slower increase in the elderly. Heart rate also increased in both groups but showed less change in the elderly. Mean blood pressure temporarily decreased in both groups but less in the elderly. The magnitude of relative change (gain) of heart rate in the elderly was significantly smaller than that in the young, whereas the increasing rate to reach one-half of the gain (response time) of ventilation in the elderly was significantly slower than that in the young. Similar tendencies were observed in the passive movements. It is concluded that the elderly show slower ventilatory response and attenuated circulatory response at the onset of dynamic voluntary exercise and passive movements. PMID- 11053326 TI - Effects of sevoflurane on the contractility of ferret ventricular myocardium. AB - Isotonic and isometric variables of contractility and relaxation of isolated ferret right ventricular papillary muscles were measured before and during exposure to incremental concentrations of sevoflurane (0-4.9% vol/vol) (30 degrees C) (n = 9). In a second group of muscles (n = 8), effects of sevoflurane were compared with those of low [Ca(2+)](o) (0.45-2.25 mM in steps of 0.45 mM). Sevoflurane caused a reversible concentration-dependent decrease in contractility (ED(50) of developed force 4.6+/-0.9% vol/vol). When compared with twitches of equal amplitude in low extracellular Ca(2+) concentration, sevoflurane accelerated both isometric and isotonic relaxation. The myocardial depressant effect of sevoflurane is less than that of isoflurane and results mainly from a decrease of intracellular Ca(2+) availability. The abbreviated isometric relaxation likely reflects a decrease in Ca(2+) sensitivity and the faster isotonic relaxation may reflect a mild stimulation of Ca(2+) uptake by the sarcoplasmic reticulum. PMID- 11053327 TI - Effect of gravity on aerosol dispersion and deposition in the human lung after periods of breath holding. AB - To determine the extent of the role that gravity plays in dispersion and deposition during breath holds, we performed aerosol bolus inhalations of 1 microm-diameter particles followed by breath holds of various lengths on four subjects on the ground (1G) and during short periods of microgravity (microG). Boluses of approximately 70 ml were inhaled to penetration volumes (V(p)) of 150 and 500 ml, at a constant flow rate of approximately 0.45 l/s. Aerosol concentration and flow rate were continuously measured at the mouth. Aerosol deposition and dispersion were calculated from these data. Deposition was independent of breath-hold time at both V(p) in microG, whereas, in 1G, deposition increased with increasing breath hold time. At V(p) = 150 ml, dispersion was similar at both gravity levels and increased with breath hold time. At V(p) = 500 ml, dispersion in 1G was always significantly higher than in microG. The data provide direct evidence that gravitational sedimentation is the main mechanism of deposition and dispersion during breath holds. The data also suggest that cardiogenic mixing and turbulent mixing contribute to deposition and dispersion at shallow V(p). PMID- 11053328 TI - Skeletal muscle metabolic and ionic adaptations during intense exercise following sprint training in humans. AB - The effects of sprint training on muscle metabolism and ion regulation during intense exercise remain controversial. We employed a rigorous methodological approach, contrasting these responses during exercise to exhaustion and during identical work before and after training. Seven untrained men undertook 7 wk of sprint training. Subjects cycled to exhaustion at 130% pretraining peak oxygen uptake before (PreExh) and after training (PostExh), as well as performing another posttraining test identical to PreExh (PostMatch). Biopsies were taken at rest and immediately postexercise. After training in PostMatch, muscle and plasma lactate (Lac(-)) and H(+) concentrations, anaerobic ATP production rate, glycogen and ATP degradation, IMP accumulation, and peak plasma K(+) and norepinephrine concentrations were reduced (P<0.05). In PostExh, time to exhaustion was 21% greater than PreExh (P<0.001); however, muscle Lac(-) accumulation was unchanged; muscle H(+) concentration, ATP degradation, IMP accumulation, and anaerobic ATP production rate were reduced; and plasma Lac(-), norepinephrine, and H(+) concentrations were higher (P<0.05). Sprint training resulted in reduced anaerobic ATP generation during intense exercise, suggesting that aerobic metabolism was enhanced, which may allow increased time to fatigue. PMID- 11053329 TI - Bronchial reactivity of healthy subjects: 18-20 h postexposure to ozone. AB - Exposure of humans to ambient levels of ozone (O(3)) causes inflammatory changes within lung tissues. These changes have been reported for the "initial" (1- to 3 h) and "late" (18- to 20-h) postexposure periods. We hypothesized that at the late period, when protein and cellular markers of inflammation at the airway surface remain abnormal and the integrity of the epithelial barrier is compromised, bronchial reactivity would be increased. To test this, we measured airway responsiveness to cumulative doses of methacholine (MCh) aerosol in healthy subjects 19+/-1 h after a single exposure to O(3) (130 min at ambient levels between 120 and 240 parts/billion and alternate periods of rest and moderate exercise) or filtered air. Exposures were conducted at two temperatures: mild (22 degrees C) and moderate (30 degrees C). At the late period, bronchial reactivity to MCh increased, i.e., interpolated dose of MCh leading to a 50% fall in specific airway conductance (PC(50)) was less after O(3) than after filtered air. PC(50) for O(3) at 22 degrees C was 27 mg/ml (20% less than the PC(50) after filtered air), and for O(3) at 30 degrees C it was 19 mg/ml (70% less than the PC(50) after filtered air). The forced expiratory volume in 1 s (FEV(1)) at the late time point after O(3) was slightly but significantly reduced (2.3%) from the preexposure level. There was no relationship found between the functional changes observed early after exposure to O(3) and subsequent changes in bronchial reactivity or FEV(1) at the late time point. These results suggest that bronchial reactivity is significantly altered approximately 1 day after O(3); this injury may contribute to the respiratory morbidity that is observed 1-2 days after an episode of ambient air pollution. PMID- 11053330 TI - Effect of L-arginine on endothelial injury and hemostasis in rabbit endotoxin shock. AB - To investigate whether impaired endothelial function was related to alteration of nitric oxide (NO) formation during endotoxic shock, we studied the effects of supplementation of L-arginine (L-Arg), D-arginine (D-Arg), and N(G)-nitro-L arginine methyl ester (L-NAME), on endothelial function and structure in a rabbit model. Endotoxic shock was induced by a single lipopolysaccharide bolus (0.5 mg/kg i.v., Escherichia coli endotoxin). Coagulation factors and expression of monocyte tissue factor were determined by functional assays. Endothelium dependent vascular relaxation was assessed by in vitro vascular reactivity. Immunohistochemical staining (CD31) was performed to assess damaged endothelial cell surface of the abdominal aorta. These parameters were studied 5 days after the onset of endotoxic shock and were compared under three conditions: in absence of treatment, with L-Arg or D-Arg supplementation, or with L-NAME. Both L-Arg and D-Arg significantly improved endothelium-dependent relaxation and endothelial morphological injury. L-NAME did not alter endothelial histological injury induced by lipopolysaccharide. These data indicate that arginine supplementation nonspecifically prevents endothelial dysfunction and histological injury in rabbit endotoxic shock. Moreover, L-Arg has no effect on coagulation activation and expression of monocyte tissue factor induced by endotoxic shock. PMID- 11053331 TI - Impaired interval exercise responses in elite female cyclists at moderate simulated altitude. AB - The effect of hypoxia on the response to interval exercise was determined in eight elite female cyclists during two interval sessions: a sustained 3 x 10-min endurance set (5-min recovery) and a repeat sprint session comprising three sets of 6 x 15-s sprints (work-to-relief ratios were 1:3, 1:2, and 1:1 for the 1st, 2nd, and 3rd sets, respectively, with 3 min between each set). During exercise, cyclists selected their maximum power output and breathed either atmospheric air (normoxia, 20.93% O(2)) or a hypoxic gas mix (hypoxia, 17.42% O(2)). Power output was lower in hypoxia vs. normoxia throughout the endurance set (244+/-18 vs. 226+/-17, 234+/-18 vs. 221+/-25, and 235+/-18 vs. 221+/-25 W for 1st, 2nd, and 3rd sets, respectively; P< 0.05) but was lower only in the latter stages of the second and third sets of the sprints (452+/-56 vs. 429+/-49 and 403+/-54 vs. 373+/- 43 W, respectively; P<0.05). Hypoxia lowered blood O(2) saturation during the endurance set (92.9+/-2.9 vs. 95.4+/-1.5%; P<0.05) but not during repeat sprints. We conclude that, when elite cyclists select their maximum exercise intensity, both sustained (10 min) and short-term (15 s) power are impaired during hypoxia, which simulated moderate ( approximately 2,100 m) altitude. PMID- 11053332 TI - Controlled 5-mo aerobic training improves heart rate but not heart rate variability or baroreflex sensitivity. AB - Endurance-trained athletes have increased heart rate variability (HRV), but it is not known whether exercise training improves the HRV and baroreflex sensitivity (BRS) in sedentary persons. We compared the effects of low- and high-intensity endurance training on resting heart rate, HRV, and BRS. The maximal oxygen uptake and endurance time increased significantly in the high-intensity group compared with the control group. Heart rate did not change significantly in the low intensity group but decreased significantly in the high-intensity group (-6 beats/min, 95% confidence interval; -10 to -1 beats/min, exercise vs. control). No significant changes occurred in either the time or frequency domain measures of HRV or BRS in either of the exercise groups. Exercise training was not able to modify the cardiac vagal outflow in sedentary, middle-aged persons. PMID- 11053333 TI - Effect of systemic nitric oxide synthase inhibition on postexercise hypotension in humans. AB - An acute bout of aerobic exercise results in a reduced blood pressure that lasts several hours. Animal studies suggest this response is mediated by increased production of nitric oxide. We tested the extent to which systemic nitric oxide synthase inhibition [N(G)-monomethyl-L-arginine (L-NMMA)] can reverse the drop in blood pressure that occurs after exercise in humans. Eight healthy subjects underwent parallel experiments on 2 separate days. The order of the experiments was randomized between sham (60 min of seated upright rest) and exercise (60 min of upright cycling at 60% peak aerobic capacity). After both sham and exercise, subjects received, in sequence, systemic alpha-adrenergic blockade (phentolamine) and L-NMMA. Phentolamine was given first to isolate the contribution of nitric oxide to postexercise hypotension by preventing reflex changes in sympathetic tone that result from systemic nitric oxide synthase inhibition and to control for alterations in resting sympathetic activity after exercise. During each condition, systemic and regional hemodynamics were measured. Throughout the study, arterial pressure and vascular resistances remained lower postexercise vs. postsham despite nitric oxide synthase inhibition (e.g., mean arterial pressure after L-NMMA was 108.0+/-2.4 mmHg postsham vs. 102.1+/-3.3 mmHg postexercise; P<0.05). Thus it does not appear that postexercise hypotension is dependent on increased production of nitric oxide in humans. PMID- 11053334 TI - Comparison of caffeine and theophylline ingestion: exercise metabolism and endurance. AB - This two-part investigation compared the ergogenic and metabolic effects of theophylline and caffeine. Initially (part A), the ergogenic potential of theophylline on endurance exercise was investigated. Eight men cycled at 80% maximum O(2) consumption to exhaustion 90 min after ingesting either placebo (dextrose), caffeine (6 mg/kg; Caff), or theophylline (4.5 mg/kg Theolair; Theo). There was a significant increase in time to exhaustion in both the Caff (41.2+/ 4.8 min) and Theo (37.4+/-5.0 min) trials compared with placebo (32.6+/-3.4 min) (P<0.05). In part B, the effects of Theo on muscle metabolism were investigated and compared with Caff. Seven men cycled for 45 min at 70% maximum O(2) consumption (identical treatment protocol as in part A). Neither methylxanthines (MX) affected muscle glycogen utilization (P>0.05). Only Caff increased plasma epinephrine (P<0.05), but both MX increased blood glycerol levels (P<0.05). Muscle cAMP was increased (P<0.05) by both MX at 15 min and remained elevated at 45 min with Theo. This demonstrates that both MX are ergogenic and that this can be independent of muscle glycogen. PMID- 11053335 TI - Preexercise carbohydrate ingestion, glucose kinetics, and muscle glycogen use: effect of the glycemic index. AB - Eight trained men cycled at 70% peak oxygen uptake for 120 min followed by a 30 min performance cycle after ingesting either a high-glycemic index (HGI), low glycemic index (LGI), or placebo (Con) meal 30 min before exercise. Ingestion of HGI resulted in an elevated (P<0.01) blood glucose concentration compared with LGI and Con. At the onset of exercise, blood glucose fell (P<0.05) such that it was lower (P<0.05) in HGI compared with LGI and Con at 15 and 30 min during exercise. Plasma insulin concentration was higher (P<0.01) throughout the rest period after ingestion of HGI compared with LGI and Con. Plasma free fatty acid concentrations were lower (P<0.05) throughout exercise in HGI compared with LGI and Con. The rates of [6,6-(2)H]glucose appearance and disappearance were higher (P<0.05) at rest after ingestion and throughout exercise in HGI compared with LGI and Con. Carbohydrate oxidation was higher (P<0.05) throughout exercise, whereas glycogen use tended (P = 0.07) to be higher in HGI compared with LGI and Con. No differences were observed in work output during the performance cycle when comparing the three trials. These results demonstrate that preexercise carbohydrate feeding with a HGI, but not a LGI, meal augments carbohydrate utilization during exercise but does not effect exercise performance. PMID- 11053336 TI - Inhaled budesonide inhibits OVA-induced airway narrowing, inflammation, and cys LT synthesis in BN rats. AB - The objective of the present investigation was to examine the effects of an inhaled glucocorticoid, budesonide, on antigen-induced production of cysteinyl leukotrienes (cys-LTs) and pulmonary inflammatory cell infiltration in the Brown Norway rat, an animal model of asthma. Two weeks after sensitization to ovalbumin, rats were treated with budesonide (2.5 mg/kg) 18 and 1 h before challenge with antigen. Budesonide abolished the late response to ovalbumin (P<0.02) and strongly inhibited the in vivo synthesis of N-acetyl-leukotriene E(4), an indicator of cys-LT synthesis, during this period (P<0.005). Both total bronchoalveolar lavage (BAL) cells (P<0.01) and BAL macrophages (P<0.005) were markedly reduced to approximately 25% of their control levels after treatment with budesonide. It can be concluded that inhibition of the antigen-induced late response in Brown Norway rats by budesonide is associated with reductions in both BAL macrophages and cys-LT synthesis. It is possible that the effect of budesonide on cys-LT synthesis is related to its effects on pulmonary macrophages. PMID- 11053337 TI - A human acinar structure for simulation of realistic alveolar plateau slopes. AB - We simulated the intra-acinar contribution to phase III slope (S(acin)) for gases of differing diffusivities (He and SF(6)) by solving equations of diffusive and convective gas transport in multi-branch-point models (MBPM) of the human acinus. We first conducted a sensitivity study of S(acin) to asymmetry and its variability in successive generations. S(acin) increases were greatest when asymmetry and variability of asymmetry were increased at the level of the respiratory bronchioles (generations 17-18) for He and at the level of the alveolar ducts (generations 20-21) for SF(6), corresponding to the location of their respective diffusion fronts. On the basis of this sensitivity study and in keeping with reported acinar morphometry, we built a MBPM that actually reproduced experimental S(acin) values obtained in normal subjects for He, N(2), and SF(6). Ten variants of such a MBPM were constructed to estimate intrinsic S(acin) variability owing to peripheral lung structure. The realistic simulation of S(acin) in the normal lung and the understanding of how asymmetry affects S(acin) for different diffusivity gases make S(acin) a powerful tool to detect structural alterations at different depths in the lung periphery. PMID- 11053338 TI - Regional blood flow during exercise in humans measured by near-infrared spectroscopy and indocyanine green. AB - Using near-infrared spectroscopy (NIRS) and the tracer indocyanine green (ICG), we quantified blood flow in calf muscle and around the Achilles tendon during plantar flexion (1-9 W). For comparison, blood flow in calf muscle was determined by dye dilution in combination with magnetic resonance imaging measures of muscle volume, and, for the peritendon region, blood flow was measured by (133)Xe washout. From rest to a peak load of 9 W, NIRS-ICG blood flow in calf muscle increased from 2.4+/-0.2 to 74+/-5 ml x 100 ml tissue(-1) x min(-1), similar to that measured by reverse dye (77+/-6 ml x 100 ml tissue(-1) x min(-1)). Achilles peritendon blood flow measured by NIRS-ICG rose with exercise from 2.2+/-0.5 to 15.1+/-0.2 ml x 100 ml(-1) x min(-1), which was similar to that determined by (133)Xe washout (2.0+/-0.6 to 14.6+/-0.3 ml x 100 ml tissue(-1) x min(-1)). This is the first study using NIRS and ICG to quantify regional tissue blood flow during exercise in humans. Due to its high spatial and temporal resolution, the technique may be useful for determining regional blood flow distribution and regulation during exercise in humans. PMID- 11053339 TI - Stability and change in genetic and environmental influences on hand-grip strength in older male twins. AB - The aim of this study was to investigate aging-related changes in the contribution of genetic and environmental influences to hand-grip strength in late adulthood. Subjects in this study are 152 intact twin pairs (77 monozygotic and 75 dizygotic pairs) from the National Heart, Lung, and Blood Institute Twin Study assessed repeatedly for hand-grip strength at mean ages of 63 and 73 yr. Structural equation genetic modeling was used to investigate stability and change in the genetic and environmental components of variance of hand-grip strength in late adulthood. Average decline in strength over the 10 yr of follow-up was 1.05+/-6.8 (SD) kg and was highly significant (P = 0.003). The test-retest correlation between baseline and follow-up grip strength was 0.62 (P<0.001). Bivariate genetic modeling found significant genetic and shared environmental stability in hand-grip strength over the 10 yr of follow-up, with genetic and shared environmental influences accounting for 35 and 48%, respectively, of the test-retest phenotypic correlation. We conclude from these results that stability in hand-grip strength in late adulthood is due primarily to continuity of genetic and familial influences. PMID- 11053340 TI - Effects of phrenicotomy and exercise on hypoxia-induced changes in phrenic motor output. AB - To investigate models of plasticity in respiratory motor output, we determined the effects of chronic unilateral phrenicotomy and/or exercise on time-dependent responses to episodic hypoxia in the contralateral phrenic nerve. Anesthetized (urethane), ventilated, and vagotomized rats were presented with three, 5-min episodes of isocapnic hypoxia (11% O(2)), separated by 5 min of hyperoxia (50% O(2)). Integrated phrenic (and hypoglossal) nerve discharge were recorded before and during each hypoxic episode, for the first 5 min after the first hypoxic episode, and at 30 and 60 min after the final episode. Of 36 rats, one-half were sedentary while the other one-half had free access to a running wheel; each of these groups was split into three subgroups: 1) unoperated, 2) chronic left phrenicotomy (27-37 days), and 3) sham operated. Neither unilateral phrenicotomy nor running wheel activity influenced the short-term hypoxic phrenic response (during hypoxia) or long-term facilitation (posthypoxia). Posthypoxia frequency decline was exaggerated in phrenicotomized-sedentary rats relative to unoperated sedentary rats (change in burst frequency = -23+/-4 vs. -11 +/-5 bursts/min, respectively; 5 min posthypoxia; P<0.05), an effect that was eliminated by spontaneous exercise. The results indicate that neither voluntary running nor unilateral phrenicotomy has major effects on time-dependent hypoxic phrenic responses, with the exception of an unexpected effect of phrenicotomy on posthypoxia frequency decline in sedentary rats. PMID- 11053341 TI - Feedforward sympathetic coronary vasodilation in exercising dogs. AB - The hypothesis that exercise-induced coronary vasodilation is a result of sympathetic activation of coronary smooth muscle beta-adrenoceptors was tested. Ten dogs were chronically instrumented with a flow transducer on the circumflex coronary artery and catheters in the aorta and coronary sinus. During treadmill exercise, coronary venous oxygen tension decreased with increasing myocardial oxygen consumption, indicating an imperfect match between myocardial blood flow and oxygen consumption. This match was improved after alpha-adrenoceptor blockade with phentolamine but was significantly worse than control after alpha + beta adrenoceptor blockade with phentolamine plus propranolol. The response after alpha-adrenoceptor blockade included local metabolic vasodilation plus a beta adrenoceptor vasodilator component, whereas the response after alpha + beta adrenoceptor blockade contained only the local metabolic vasodilator component. The large difference in coronary venous oxygen tensions during exercise between alpha-adrenoceptor blockade and alpha + beta-adrenoceptor blockade indicates that there is significant feedforward beta-adrenoceptor coronary vasodilation in exercising dogs. Coronary venous and estimated myocardial interstitial adenosine concentrations did not increase during exercise before or after alpha + beta adrenoceptor blockade, indicating that adenosine levels did not increase to compensate for the loss of feedforward beta-adrenoceptor-mediated coronary vasodilation. These results indicate a meaningful role for feedforward beta receptor-mediated sympathetic coronary vasodilation during exercise. PMID- 11053342 TI - Quantitative analysis of feedforward sympathetic coronary vasodilation in exercising dogs. AB - Recent experiments demonstrate that feedforward sympathetic beta-adrenoceptor coronary vasodilation occurs during exercise. The present study quantitatively examined the contributions of epinephrine and norepinephrine to exercise coronary hyperemia and tested the hypothesis that circulating epinephrine causes feedforward beta-receptor-mediated coronary dilation. Dogs (n = 10) were chronically instrumented with a circumflex coronary artery flow transducer and catheters in the aorta and coronary sinus. During strenuous treadmill exercise, myocardial oxygen consumption increased by approximately 3.9-fold, coronary blood flow increased by approximately 3.6-fold, and arterial plasma epinephrine concentration increased by approximately 2.4-fold over resting levels. At arterial concentrations matching those during strenuous exercise, epinephrine infused at rest (n = 6) produced modest increases (18%) in flow and myocardial oxygen consumption but no evidence of direct beta-adrenoceptor-mediated coronary vasodilation. Arterial norepinephrine concentration increased by approximately 5. 4-fold during exercise, and coronary venous norepinephrine was always higher than arterial, indicating norepinephrine release from cardiac sympathetic nerves. With the use of a mathematical model of cardiac capillary norepinephrine transport, these norepinephrine concentrations predict an average interstitial norepinephrine concentration of approximately 12 nM during strenuous exercise. Published dose-response data indicate that this norepinephrine concentration increases isolated coronary arteriolar conductance by approximately 67%, which can account for approximately 25% of the increase in flow observed during exercise. It is concluded that a significant portion of coronary exercise hyperemia ( approximately 25%) can be accounted for by direct feedforward beta adrenoceptor coronary vascular effects of norepinephrine, with little effect from circulating epinephrine. PMID- 11053343 TI - Total power output generated during dynamic knee extensor exercise at different contraction frequencies. AB - A novel approach has been developed for the quantification of total mechanical power output produced by an isolated, well-defined muscle group during dynamic exercise in humans at different contraction frequencies. The calculation of total power output comprises the external power delivered to the ergometer (i.e., the external power output setting of the ergometer) and the "internal" power generated to overcome inertial and gravitational forces related to movement of the lower limb. Total power output was determined at contraction frequencies of 60 and 100 rpm. At 60 rpm, the internal power was 18+/- 1 W (range: 16-19 W) at external power outputs that ranged between 0 and 50 W. This was less (P<0.05) than the internal power of 33+/-2 W (27-38 W) at 100 rpm at 0-50 W. Moreover, at 100 rpm, internal power was lower (P<0.05) at the higher external power outputs. Pulmonary oxygen uptake was observed to be greater (P<0.05) at 100 than at 60 rpm at comparable total power outputs, suggesting that mechanical efficiency is lower at 100 rpm. Thus a method was developed that allowed accurate determination of the total power output during exercise generated by an isolated muscle group at different contraction frequencies. PMID- 11053344 TI - State influences on ventral medullary surface and physiological responses to sodium cyanide challenges. AB - Intravenous sodium cyanide (NaCN) administration lowers ventral medullary surface (VMS) activity in anesthetized cats. Sleep states modify spontaneous and blood pressure-evoked VMS activity and may alter VMS responses to chemoreceptor input. We studied VMS activation during peripheral chemoreceptor stimulation by intravenous NaCN using optical procedures in six cats instrumented for recording sleep physiology during sham saline and control site trials. Images of scattered 660-nm light were collected at 50 frames/s with an optical device after 80-100 microg total bolus intravenous NaCN delivery during waking and sleep states. Cyanide elicited an initial ventilatory decline, followed by large inspiratory efforts and an increase in respiratory rate, except in rapid eye movement sleep, in which an initial breathing increase occurred. NaCN evoked a pronounced decrease in VMS activity in all states; control sites and sham injections showed little effect. The activity decline was faster in rapid eye movement sleep, and the activity nadir occurred later in waking. Sleep states alter the time course but not the extent of decline in VMS activity. PMID- 11053345 TI - Downhill running preferentially increases CGRP in fast glycolytic muscle fibers. AB - Calcitonin gene-related peptide (CGRP) is present in some spinal cord motoneurons and at neuromuscular junctions in skeletal muscle. We previously reported increased numbers of CGRP-positive (CGRP+) motoneurons supplying hindlimb extensors after downhill exercise (Homonko DA and Theriault E, Inter J Sport Med 18: 1-7, 1997). The present study identifies the responding population with respect to muscle and motoneuron pool and correlates changes in CGRP with muscle fiber type-identified end plates. Twenty seven rats were divided into the following groups: control and 72 h and 2 wk postexercise. FluoroGold was injected into the soleus, lateral gastrocnemius, and the proximal (mixed fiber type) or distal (fast-twitch glycolytic) regions of the medial gastrocnemius (MG). Untrained animals ran downhill on a treadmill for 30 min. The number of FluoroGold/CGRP+ motoneurons within proximal and distal MG increased by 72 h postexercise (P<0.05). No significant changes were observed in soleus or lateral gastrocnemius motoneurons postexercise. The number of alpha-bungarotoxin/CGRP+ motor end plates in the MG increased exclusively at fast-twitch glycolytic muscle fibers 72 h and 2 wk postexercise (P<0.05). One interpretation of these results is that unaccustomed exercise preferentially activates fast-twitch glycolytic muscle fibers in the MG. PMID- 11053346 TI - Expression of hypoxia-inducible factor-1alpha in the brain of rats during chronic hypoxia. AB - Hypoxia-inducible factor-1 (HIF-1) is a transcription factor that regulates adaptive responses to the lack of oxygen in mammalian cells. HIF-1 consists of two proteins, HIF-1alpha and HIF-1beta. HIF-1alpha accumulates under hypoxic conditions, whereas HIF-1beta is constitutively expressed. HIF-1alpha and HIF 1beta expression were measured during adaptation to hypobaric hypoxia (0.5 atm) in rat cerebral cortex. Western blot analyses indicated that HIF-1alpha rapidly accumulated during the onset of hypoxia and did not fall for 14 days but fell to normal by 21 days despite the continuous low arterial oxygen tension. Immunostaining showed that neurons, astrocytes, ependymal cells, and possibly endothelial cells were the cell types expressing HIF-1alpha. Genes with hypoxia responsive elements were activated under these conditions, as evidenced by elevated vascular endothelial growth factor and glucose transporter-1 mRNA levels. When 21-day-adapted rats were exposed to a more severe hypoxic challenge (8% oxygen), HIF-1alpha accumulated again. On the basis of these results, we speculate that the vascular remodeling and metabolic changes triggered during prolonged hypoxia are capable of restoring normal tissue oxygen levels. PMID- 11053347 TI - Role of positive airway pressure on pulmonary acinar perfusion heterogeneity. AB - Perfusion of the pulmonary acinus has been shown to be generally homogeneous, but there is a significant component that is heterogeneous. To investigate the contribution of the alveolar septal capillary network to acinar perfusion heterogeneity, the passage of fluorescent dye boluses through the subpleural microcirculation of isolated dog lung lobes was videotaped using fluorescence microscopy. As the videotapes were replayed, dye-dilution curves were recorded from each of the tributary branches of Y-shaped venules that drained single acini. For each Y-shaped venule, the mean appearance time difference between the pair of tributary branches was calculated from the dye curves. When the complex septal capillary networks were derecruited by high positive airway pressure, venular perfusion became proportionally more homogeneous. This result shows that septal capillary resistance and pathlength differences are important contributors to intra-acinar perfusion heterogeneity. PMID- 11053348 TI - Eicosanoid and muscarinic receptor blockade abolishes hyperventilation-induced bronchoconstriction. AB - This study was designed to test the hypothesis that hyperventilation-induced bronchoconstriction (HIB) results from the combined effects of prostanoid and leukotriene metabolism. A bronchoscope was used in anesthetized dogs to record peripheral airway resistance and HIB before and after combined treatment with inhibitors of cyclooxygenase (indomethacin) and 5-lipoxygenase (MK-0591). Bronchoalveolar lavage fluid (BALF) cells and mediators from hyperventilated and control airways were also measured. Pretreatment with MK-0591 and indomethacin significantly attenuated, but did not abolish, HIB. However, addition of atropine nearly eliminated the residual response. Blockade of eicosanoid metabolism markedly reduced the concentrations of eicosanoids recovered in BALF after hyperventilation. Positive correlations between posthyperventilation BALF prostanoid and epithelial cell concentrations are suggestive of mucosal injury induced mediator production and release. We conclude that HIB is prevented in the presence of eicosanoid and muscarinic-receptor blockade and that both classes of eicosanoids contribute similarly to the development of HIB. PMID- 11053349 TI - Arterial properties of the carotid and femoral artery in endurance-trained and paraplegic subjects. AB - In humans, the relationships of blood flow changes to structure, function, and shear rate of conducting arteries have not been thoroughly examined. Therefore, the purpose of this study was to investigate these parameters of the elastic type, common carotid artery (CCA) and the muscular-type, common femoral artery (CFA) in long-term highly active and extremely inactive individuals, assuming that the impact of activity-induced blood flow changes on conduit arteries, if any, should be seen in these subjects. We examined 21 highly endurance-trained athletes (A), 10 paraplegic subjects (P), and 20 sedentary subjects (S) by means of noninvasive ultrasound. As a result, the CFA diameter and compliance were highest in A (9.7+/-0.81 mm; 1.84 +/-0.54 mm(2)/kPa) and lowest in P (5.9+/-0.7 mm; 0.54+/-0.27 mm(2)/kPa) compared with S (8.3+/-1.0 mm; 0.92+/-0.48 mm(2)/kPa) with P <0.01 among the groups. Both parameters correlated with each other (r = 0.62; P<0.01). Compared with A (378+/-84 s(-1); 37+/-15 s(-1)) and S (356+/-113 s(-1); 36+/-20 s(-1)), the peak and mean shear rates of the CFA were almost or more than doubled in P (588+/-120 s(-1); 89+/-26 s(-1)). In the CCA, only the compliance and peak shear rate showed significant differences among the groups (A: 1.28+/-0.47 mm(2)/kPa, 660+/-138 s(-1); S: 1.04+/-0.27 mm(2)/kPa, 588+/-109 s(-1); P: 0.65+/- 0.22 mm(2)/kPa, 490+/-149 s(-1); P<0.05). In conclusion, the results suggest a structural and functional adaptation in the CFA and a predominantly functional adaptation of the arterial wall properties to differences in the physical activity level and associated exercise-induced blood flow changes in the CCA. The results for humans confirm those from animal experiments. Similar shear rate values of S and P in the CFA support the hypothesis of constant shear stress regulation due to local blood flow changes in humans. On the other hand, the increased shear rate in the CFA in P indicates an at least partially nonphysiological response of the arterial wall in long-term chronic sympathectomy due to a change in local blood flow. PMID- 11053350 TI - Submandibular secretory and vascular responses to stimulation of the parasympathetic innervation in anesthetized cats. AB - Submandibular secretory responses to stimulation of the parasympathetic chorda lingual nerve in anaesthetized cats have been investigated before, during, and after intracarotid infusion of endothelin-1 (ET-1), which reduced blood flow through the gland by 64+/-7%. Stimulation at different frequencies (2, 4, 8, and 16 Hz) evoked a frequency-dependent increase in the flow of submandibular saliva, sodium concentration and output, and output of both potassium and protein. The reduction in submandibular blood flow, which occurred in response to the infusion of ET-1, was associated with a decreased flow of saliva and a diminished output of both sodium and protein. The flow of saliva was linearly related to submandibular blood flow both in the presence and absence of ET-1. It is concluded that submandibular secretory responses to electrical stimulation of the parasympathetic innervation can be significantly attenuated by reducing the blood flow through the gland by ET-1 infusion, just as it is when the blood flow is reduced by hypotension. PMID- 11053351 TI - Airway function after cyclooxygenase inhibition during hyperpnea-induced bronchoconstriction in guinea pigs. AB - Airway function deteriorates significantly on cessation of exercise or isocapnic hyperventilation challenges but is largely preserved during the challenge in humans and guinea pigs. PGE(2), an endogenous bronchodilator, might be responsible for the preservation of lung function during hyperventilation (HV). We hypothesized that PGE(2) might have a protective effect during HV, partially explaining the minimal changes in respiratory system resistance (Rrs) usually seen during HV in humans and guinea pigs. Therefore, changes in Rrs were measured during and after HV in anesthetized, mechanically ventilated guinea pigs treated with flurbiprofen (FBN) or placebo. With HV, there was an initial bronchodilation that was unaffected by FBN. Rrs then increased with time during HV, an effect that was blocked by FBN. After HV, Rrs increased further in all groups, but the increase in Rrs was less in the FBN-treated groups. FBN treatment reduced the PGE(2) concentration slightly in lung lavage fluid compared with placebo. We found no enhancement or refractoriness of the Rrs response to repeat bouts of HV and no effect of FBN treatment on the response of Rrs to repeat HV. These results suggest that a constrictor PG is released during and possibly after HV and that the post-HV increase in Rrs is the sum of effects of the PG released during HV and a second constrictor mechanism operating after HV. We found no evidence for bronchodilator PG during or after HV in the guinea pig. PMID- 11053352 TI - Counteraction of aortic baroreflex to carotid sinus baroreflex in a neck suction model. AB - Although neck suction has been widely used in the evaluation of carotid sinus baroreflex function in humans, counteraction of the aortic baroreflex tends to complicate any interpretation of observed arterial pressure (AP) response. To determine whether a simple linear model can account for the AP response during neck suction, we developed an animal model of the neck suction procedure in which changes in carotid distension pressure during neck suction were directly imposed on the isolated carotid sinus. In six anesthetized rabbits, a 50-mmHg pressure perturbation on the carotid sinus decreased AP by -27.4+/-4.8 mmHg when the aortic baroreflex was disabled. Enabling the aortic baroreflex significantly attenuated the AP response (-21.5+/-3.8 mmHg, P<0.01). The observed closed-loop gain during simulated neck suction was well predicted by the open-loop gains of the carotid sinus and aortic baroreflexes using the linear model (-0.43+/-0.13 predicted vs. -0.41 +/-0.10 measured). We conclude that the linear model can be used as the first approximation to interpret AP response during neck suction. PMID- 11053353 TI - Mutagenesis analysis of human SM22: characterization of actin binding. AB - SM22 is a 201-amino acid actin-binding protein expressed at high levels in smooth muscle cells. It has structural homology to calponin, but how SM22 binds to actin remains unknown. We performed site-directed mutagenesis to generate a series of NH(2)-terminal histidine (His)-tagged mutants of human SM22 in Escherichia coli and used these to analyze the functional importance of potential actin binding domains. Purified full-length recombinant SM22 bound to actin in vitro, as demonstrated by cosedimentation assay. Binding did not vary with calcium concentration. The COOH-terminal domain of SM22 is required for actin affinity, because COOH terminally truncated mutants [SM22-(1-186) and SM22-(1-166)] exhibited markedly reduced cosedimentation with actin, and no actin binding of SM22-(1-151) could be detected. Internal deletion of a putative actin binding site (154-KKAQEHKR-161) partially prevented actin binding, as did point mutation to neutralize either or both pairs of positively charged residues at the ends of this region (KK154LL and/or KR160LL). Internal deletion of amino acids 170-180 or 170-186 also partially or almost completely inhibited actin cosedimentation, respectively. Of the three consensus protein kinase C or casein kinase II phosphorylation sites in SM22, only Ser-181 was readily phosphorylated by protein kinase C in vitro, and such phosphorylation greatly decreased actin binding. Substitution of Ser-181 to aspartic acid (to mimic serine phosphorylation) also reduced actin binding. Immunostains of transiently transfected airway myocytes revealed that full-length NH(2)-terminal FLAG-tagged SM22 colocalizes with actin filaments, whereas FLAG-SM22-(1-151) does not. These data confirm that SM22 binds to actin in vitro and in vivo and, for the first time, demonstrate that multiple regions within the COOH-terminal domain are required for full actin affinity. PMID- 11053354 TI - Faster top running speeds are achieved with greater ground forces not more rapid leg movements. AB - We twice tested the hypothesis that top running speeds are determined by the amount of force applied to the ground rather than how rapidly limbs are repositioned in the air. First, we compared the mechanics of 33 subjects of different sprinting abilities running at their top speeds on a level treadmill. Second, we compared the mechanics of declined (-6 degrees ) and inclined (+9 degrees ) top-speed treadmill running in five subjects. For both tests, we used a treadmill-mounted force plate to measure the time between stance periods of the same foot (swing time, t(sw)) and the force applied to the running surface at top speed. To obtain the force relevant for speed, the force applied normal to the ground was divided by the weight of the body (W(b)) and averaged over the period of foot-ground contact (F(avge)/W(b)). The top speeds of the 33 subjects who completed the level treadmill protocol spanned a 1.8-fold range from 6.2 to 11.1 m/s. Among these subjects, the regression of F(avge)/W(b) on top speed indicated that this force was 1.26 times greater for a runner with a top speed of 11.1 vs. 6.2 m/s. In contrast, the time taken to swing the limb into position for the next step (t(sw)) did not vary (P = 0.18). Declined and inclined top speeds differed by 1.4-fold (9.96+/-0.3 vs. 7.10+/-0.3 m/s, respectively), with the faster declined top speeds being achieved with mass-specific support forces that were 1.3 times greater (2.30+/- 0.06 vs. 1.76+/-0.04 F(avge)/ W(b)) and minimum t(sw) that were similar (+8%). We conclude that human runners reach faster top speeds not by repositioning their limbs more rapidly in the air, but by applying greater support forces to the ground. PMID- 11053355 TI - Gastrointestinal tract, hepatic, hindlimb, and renal recovery of CO(2) in vivo. AB - Whole body oxidative rates of labeled substrates are often measured by collecting expired air and determining the amount of labeled CO(2) that is produced. However, the CO(2) produced may not be completely recovered under all circumstances, and there is a wide variation in values reported under different experimental conditions ( approximately 50-100%). The potential contribution of specific organs to this variation has not been defined. In vivo studies using healthy, postabsorptive, multicatheterized conscious canines were conducted to determine gastrointestinal tract, hepatic, hindlimb, and renal recoveries of NaH(14)CO(3) during a 180-min constant infusion [0.022+/-0.002 (SE) microCi x kg( 1) x min(-1)]. Before the constant infusion period, a bolus infusion of NaH(14)CO(3) (1.76+/-0.16 microCi/kg) was given, and the rate of decay in blood was measured over a 15-min period to determine pool size. The pool size for the distribution of (14)CO(2) was approximately 80% of the total body pool (16.0+/ 1.7 liters). Whole body recovery was 97.2+/-6.7%. The recoveries across the liver, gut, leg, and kidney were 99.9+/-1.3, 98.0 +/-1.4, 96.7+/-2.6, and 99.9+/ 2.1%, respectively. In conclusion, hepatic, gastrointestinal tract, hindlimb, and renal recoveries of CO(2) in vivo were near 100%, suggesting that CO(2) loss is not greater in gluconeogenic organs and that corrections for incomplete recovery of CO(2), when measuring oxidation of substrates across these organs under normal postabsorptive conditions, would be very minor. PMID- 11053356 TI - Effect of expiratory resistive loading on the noninvasive tension-time index in COPD. AB - Expiratory resistive loading (ERL) is used by chronic obstructive pulmonary disease (COPD) patients to improve respiratory function. We, therefore, used a noninvasive tension-time index of the inspiratory muscles (TT(mus) = I/PI(max) x TI/TT, where I is mean inspiratory pressure estimated from the mouth occlusion pressure, PI(max) is maximal inspiratory pressure, TI is inspiratory time, and TT is total respiratory cycle time) to better define the effect of ERL on COPD patients. To accomplish this, we measured airway pressures, mouth occlusion pressure, respiratory cycle flow rates, and functional residual capacity (FRC) in 14 COPD patients and 10 normal subjects with and without the application of ERL. TT(mus) was then calculated and found to drop in both COPD and normal subjects (P<0.05). The decline in TT(mus) in both groups resulted solely from a prolongation of expiratory time with ERL (P<0.001 for COPD, P<0.05 for normal subjects). In contrast to the COPD patients, normal subjects had an elevation in I and FRC, thus minimizing the decline in TT(mus). In conclusion, ERL reduces the potential for inspiratory muscle fatigue in COPD by reducing TI/TT without affecting FRC and I. PMID- 11053357 TI - Aminophylline modulation of the mouse respiratory network changes during postnatal maturation. AB - Aminophylline is a respiratory stimulant commonly used for the treatment of central apnea. Experiences from clinical practice, however, revealed that aminophylline is not reliably effective in preterm infants, whereas it is normally effective in infants and mature patients. In an established animal model for postnatal development of respiratory control mechanisms, we therefore examined the hypothesis that the clinical observations reflect a developmental change in the sensitivity of the central respiratory network to methylxanthines. The medullary respiratory network was isolated at different postnatal ages (postnatal days 1-13; P1-P13) in a transverse mouse brain stem slice preparation. This preparation contains the pre-Botzinger complex (PBC), a region that is critical for generation of respiratory rhythm. Spontaneous rhythmic respiratory activity was recorded from the hypoglossal (XII) rootlets and from neurons in the PBC by using the whole cell patch clamp technique. Bath-applied aminophylline [20 microM] increased the frequency (+41%) in neonatal animals (P1-P6) without affecting the amplitude of respiratory burst activity in XII rootlets. The same concentration of aminophylline did not have any significant effect on the frequency of respiratory XII bursts but increased the amplitude (+31%) in juvenile animals (P7-P13). In the same age group, aminophylline also augmented the amplitude and the duration of respiratory synaptic drive currents in respiratory PBC neurons. The data demonstrate that augmentation of the respiratory output is due to direct enhancement of central respiratory network activity and increase of synaptic drive of hypoglossal motoneurons in juvenile, but not neonatal, animals. This indicates a developmental change in the efficacy of aminophylline to reinforce central respiratory network activity. Therefore, we believe that the variable success in treating respiratory disturbances in premature infants reflects maturational changes in the expression of receptors and/or intracellular signal pathways in the central respiratory network. PMID- 11053358 TI - Kinetics of respiratory system elastance after airway challenge in dogs. AB - We compared the time courses of lung mechanical changes with intravenous (iv) injection vs. aerosol administration of histamine, methacholine, and ACh in dogs. We interpret these results in terms of a spring-and-dashpot model of airway smooth muscle receiving activation via a tissue compartment when agonist is delivered by the iv route and through an additional airway wall compartment when it is delivered by the aerosol route. The model accurately accounts for the principal features of the respiratory system elastance response curves. It also accounts for the differences between iv and aerosol responses, supporting the notion that agonist delivered by aerosol has to traverse a longer pathway to the airway smooth muscle than does agonist delivered iv. The time constants representing diffusive exchange of agonist between compartments were not significantly different for the three agonists, suggesting that the three agonists shared a common principal means of clearance, which was presumably blood flow. PMID- 11053359 TI - Size distribution of recruited alveolar volumes in airway reopening. AB - In 11 isolated dog lung lobes, we studied the size distribution of recruited alveolar volumes that become available for gas exchange during inflation from the collapsed state. Three catheters were wedged into 2-mm-diameter airways at total lung capacity. Small-amplitude pseudorandom pressure oscillations between 1 and 47 Hz were led into the catheters, and the input impedances of the regions subtended by the catheters were continuously recorded using a wave tube technique during inflation from -5 cm H(2)O transpulmonary pressure to total lung capacity. The impedance data were fit with a model to obtain regional tissue elastance (Eti) as a function of inflation. First, Eti was high and decreased in discrete jumps as more groups of alveoli were recruited. By assuming that the number of opened alveoli is inversely proportional to Eti, we calculated from the jumps in Eti the distribution of the discrete increments in the number of opened alveoli. This distribution was in good agreement with model simulations in which airways open in cascade or avalanches. Implications for mechanical ventilation may be found in these results. PMID- 11053360 TI - Influence of dual ET(A)/ET(B)-receptor blockade on coronary responses to treadmill exercise in dogs. AB - We hypothesized that endothelin (ET) release during exercise may be triggered by alpha-adrenergic-receptor activation and thereby influence coronary hemodynamics and O(2) metabolism in dogs. Exercise resulted in coronary blood flow increases (to 1.88+/-0.26 from 1.10+/- 0.12 ml x min(-1) x g(-1)) and in a fall (P<0.01) in coronary sinus O(2) saturation (17.4+/-1.5 to 9.6+/-0.7 vol%), whereas myocardial O(2) consumption (MVO(2)) increased (109+/-13% from 145+/-16 microl O(2) min(-1) x g(-1)). Tezosentan, a dual ET(A)/ET(B)-receptor blocker, slightly reduced mean arterial pressure (MAP) and increased heart rate throughout exercise. The relationship between coronary sinus O(2) saturation and MVO(2) was shifted upward (P<0.05) after tezosentan administration; i.e., as MVO(2) increased during exercise, coronary sinus O(2) saturation was disproportionately higher after ET receptor blockade. After propranolol, tezosentan resulted in significant decreases (P<0.05) in left ventricular pressure, the first derivative of left ventricular pressure over time, and MAP during exercise. As MVO(2) increased during exercise, coronary sinus O(2) saturation levels after tezosentan became superimposable over those observed before ET-receptor blockade. Thus dual blockade of ET(A)/ET(B) receptors alters coronary hemodynamics and O(2) metabolism during exercise, but ET activity failed to increase beyond baseline levels. PMID- 11053361 TI - Postprandial lipemia in young men and women of contrasting training status. AB - This study compared the postprandial triacylglycerol (TAG) response to a high-fat meal in trained and untrained normolipidemic young adults after 2 days' abstinence from exercise. Fifty-three subjects (11 endurance-trained men, 9 endurance-trained women, 10 sprint/strength-trained men, 11 untrained men, 11 untrained women) consumed a meal (1.2 g fat, 1.1 g carbohydrate, 66 kJ per kg body mass) after a 12-h fast. Venous blood samples were obtained in the fasted state and at intervals until 6 h. Postprandial responses were the areas under the plasma or serum concentration-vs.-time curves. Neither fasting TAG concentrations nor the postprandial TAG response differed between trained and untrained subjects. The insulinemic response was 29% lower in endurance-trained men than in untrained men [mean difference -37.4 (95% confidence interval -62.9 to -22.9) microIU/ml x h, P = 0.01]. Responses of plasma glucose, serum insulin, and plasma nonesterified fatty acids were all lower for endurance-trained men than for untrained men. These findings suggest that, in young adults, no effect of training on postprandial lipemia can be detected after 60 h without exercise. The effect on postprandial insulinemia may persist for longer. PMID- 11053362 TI - Kinetics of intramuscular triglyceride fatty acids in exercising humans. AB - A pulse ([(14)C]palmitate)-chase ([(3)H]palmitate) approach was used to study intramuscular triglyceride (imTG) fatty acid and plasma free fatty acid (FFA) kinetics during exercise at approximately 45% peak O(2) consumption in 12 adults. Vastus lateralis muscle was biopsied before and after 90 min of bicycle exercise; (3)H(2)O production, breath (14)CO(2) excretion and lipid oxidation (indirect calorimetry) rates were measured during exercise. RESULTS: during exercise, 8.2+/ 1.2 and 8.4+/-0.7 micromol x kg(-1) x min(-1) of imTG fatty acids and plasma FFA, respectively, were oxidized according to isotopic measurements. The sum of these two values was not different (P = 0.6) from lipid oxidation by indirect calorimetry (15.4 +/-1.6 micromol x kg(-1) x min(-1)); the isotopic and indirect calorimetry values were correlated (r = 0.79, P<0.005). During exercise, imTG turnover rate was 0.32+/-0.07%/min (6.0+/-2.0 micromol of imTG x kg wet muscle( 1) x min(-1)) and plasma FFA were incorporated into imTG at a rate of 0.7+/-0.1 micromol x kg wet muscle(-1) x min(-1). The imTG pool size did not change during exercise. This pulse-chase, dual tracer appears to be a reasonable approach to measure oxidation and synthesis kinetics of imTG. PMID- 11053363 TI - Response of arterial smooth muscle to length perturbation. AB - The ability of arterial smooth muscle to generate tension is influenced by muscle length. An unsettled question is whether the length-tension relationship is a simple reflection of the contractile filament overlap, as it is in skeletal muscle. There are several factors that could potentially affect tension generation in arterial smooth muscle; these include stretch-induced myogenic response and length-oscillation-induced disruption of the contractile filament organization. In this study, in which rabbit carotid arterial preparations were used, we found that different length-tension curves could be obtained at different times after a length change. In addition, length oscillation at a frequency of normal pulse rate and with small to moderate oscillation amplitude was found to potentiate tension generation but reduced tension at large amplitudes. The observed response could be attributed to adaptation of the muscle to length change over time and to myogenic potentiation associated with stretching of the muscle. PMID- 11053364 TI - The effects of hindlimb unweighting on the capacitance of rat small mesenteric veins. AB - Microgravity is associated with an impaired cardiac output response to orthostatic stress. Mesenteric veins are critical in modulating cardiac filling through venoconstriction. The purpose of this study was to determine the effects of simulated microgravity on the capacitance of rat mesenteric small veins. We constructed pressure-diameter relationships from vessels of 21-day hindlimb unweighted (HLU) rats and control rats by changing the internal pressure and measuring the external diameter. Pressure-diameter relationships were obtained both before and after stimulation with norepinephrine (NE). The pressure-diameter curves of HLU vessels were shifted to larger diameters than control vessels. NE (10(-4) M) constricted veins from control animals such that the pressure-diameter relationship was significantly shifted downward (i.e., to smaller diameters at equal pressure). NE had no effect on vessels from HLU animals. These results indicate that, after HLU, unstressed vascular volume may be increased and can no longer decrease in response to sympathetic stimulation. This may partially underlie the mechanism leading to the exaggerated fall in cardiac output and stroke volume seen in astronauts during an orthostatic stress after exposure to microgravity. PMID- 11053365 TI - Invited review: mechanochemical signal transduction in the fetal lung. AB - Growth and maturation of fetal lungs are regulated by both humoral and physical factors. Mechanical stretch stimulates fetal lung cell proliferation and affects fetal lung maturation by influencing the production of extracellular matrix molecules and the expression of specific genes of fetal lung cells. These effects are mediated through special signal transduction pathways in fetal lung cells. Various in vivo and in vitro model systems have been developed to investigate the mechanotransduction process. The diversity and discrepancy of these studies have raised many questions. We will briefly summarize mechanical force-induced signals in fetal lung cell proliferation and differentiation and then discuss several important issues related to these studies. PMID- 11053367 TI - commentary PMID- 11053366 TI - Invited review: engineering approaches to cytoskeletal mechanics. AB - An outstanding problem in cell biology is how cells sense mechanical forces and how those forces affect cellular functions. Various biophysical and biochemical mechanisms have been invoked to answer this question. A growing body of evidence indicates that the deformable cytoskeleton (CSK), an intracellular network of interconnected filamentous biopolymers, provides a physical basis for transducing mechanical signals into biochemical signals. Therefore, to understand how mechanical forces regulate cellular functions, it is important to know how cells respond to changes in the CSK force balance and to identify the underlying mechanisms that control transmission of mechanical forces throughout the CSK and bring it to equilibrium. Recent developments of new experimental techniques for measuring cell mechanical properties and novel theoretical models of cellular mechanics make it now possible to identify and quantitate the contributions of various CSK structures to the overall balance of mechanical forces in the cell. This review focuses on engineering approaches that have been used in the past two decades in studies of the mechanics of the CSK. PMID- 11053368 TI - Selected contribution: mechanical strain increases force production and calcium sensitivity in cultured airway smooth muscle cells. AB - Cultured airway smooth muscle cells subjected to cyclic deformational strain have increased cell content of myosin light chain kinase (MLCK) and myosin and increased formation of actin filaments. To determine how these changes may increase cell contractility, we measured isometric force production with changes in cytosolic calcium in individual permeabilized cells. The pCa for 50% maximal force production was 6.6+/-0.4 in the strain cells compared with 5.9+/-0.3 in control cells, signifying increased calcium sensitivity in strain cells. Maximal force production was also greater in strain cells (8.6+/-2.9 vs. 5.7+/-3.1 microN). The increased maximal force production in strain cells persisted after irreversible thiophosphorylation of myosin light chain, signifying that increased force could not be explained by differences in myosin light chain phosphorylation. Cells strained for brief periods sufficient to increase cytoskeletal organization but insufficient to increase contractile protein content also produced more force, suggesting that strain-induced cytoskeletal reorganization also increases force production. PMID- 11053369 TI - Selected contribution: axial stretch increases spontaneous pacemaker activity in rabbit isolated sinoatrial node cells. AB - Isolated, spontaneously beating rabbit sinoatrial node cells were subjected to longitudinal stretch, using carbon fibers attached to both ends of the cell. Their electrical behavior was studied simultaneously in current-clamp or voltage clamp mode using the perforated patch configuration. Moderate stretch ( approximately 7%) caused an increase in spontaneous beating rate (by approximately 5%) and a reduction in maximum diastolic and systolic potentials (by approximately 2.5%), as seen in multicellular preparations. Mathematical modeling of the stretch intervention showed the experimental results to be compatible with stretch activation of cation nonselective ion channels, similar to those found in other cardiac cell populations. Voltage-clamp experiments validated the presence of a stretch-induced current component with a reversal potential near -11 mV. These data confirm, for the first time, that the positive chronotropic response of the heart to stretch is, at least in part, encoded on the level of individual sinoatrial node pacemaker cells; all reported data are in agreement with a major contribution of stretch-activated cation nonselective channels to this response. PMID- 11053370 TI - The histidine kinase domain of UhpB inhibits UhpA action at the Escherichia coli uhpT promoter. AB - The histidine kinase (HK) component of many two-component regulatory systems exhibits regulated ability to phosphorylate itself and to participate in transfer of phosphate to and from its cognate response regulator. The signaling system that controls expression of the UhpT sugar phosphate transporter in Escherichia coli in response to external glucose 6-phosphate includes the HK protein UhpB and the polytopic membrane protein UhpC, a UhpT homolog which is required for responsiveness to an inducer and activation of UhpB. The existence of a UhpBC signaling complex is suggested by the requirement for UhpC for the activity of certain constitutively active variants of UhpB, the dominance and epistasis relationships of uhp alleles, and the finding that expression of UhpB in excess of UhpC has a strong dominant-negative effect. Expression of a hybrid protein containing the cytoplasmic C-terminal half of UhpB fused to glutathione S transferase (GST) also interfered with Uhp signaling. This interference phenotype could not result solely from the phosphatase activity of UhpB, because interference affected both overexpressed UhpA and UhpA variants which are active in the absence of phosphorylation. Variant forms of UhpB which were active in the absence of UhpC carried amino acid substitutions near motifs conserved in HK proteins. The GST fusion protein inhibited the ability of UhpA to bind and activate transcription at the uhpT promoter. Unlike the wild-type situation, a GST fusion variant carrying one of the UhpB-activating substitutions, R324C, displayed autokinase activity and phosphate transfer to UhpA but retained the ability to sequester UhpA when it was altered in the conserved residues important for phosphate transfer. Thus, the default state of UhpB is kinase off, and activation of its phosphate transfer activity requires either the action of UhpC or the occurrence of certain mutations in UhpB. The interference phenotype shown by UhpB in excess of UhpC appears to include the binding and sequestration of UhpA. PMID- 11053371 TI - radC102 of Escherichia coli is an allele of recG. AB - The radC102 mutation causes mild UV and X-ray sensitivity and was mapped previously to near pyrE and recG at 82 min on the Escherichia coli chromosome (I. Felzenszwalb, N. J. Sargentini, and K. C. Smith, Radiat. Res. 97:615-625, 1984). We report that radC102 has two striking phenotypes characteristic of recG mutations. First, it causes dramatically increased RecA-dependent mutation in a stationary-phase mutation assay. Second, it causes extreme UV sensitivity in combination with ruv mutations affecting the RuvABC Holliday junction resolution system. DNA sequencing of the radC and recG genes in radC102 strains revealed that the radC102 mutation creates a stop codon in recG that is predicted to truncate the RecG protein at 410 of 603 amino acids. A low-copy-number plasmid carrying the radC(+) gene did not affect the UV sensitivity of a wild-type strain, a radC102 strain, or a recG258::Tn10mini-kan strain. We conclude that radC102 is an allele of recG and that the function of the RadC protein remains to be determined. PMID- 11053372 TI - Maltose and maltodextrin transport in the thermoacidophilic gram-positive bacterium Alicyclobacillus acidocaldarius is mediated by a high-affinity transport system that includes a maltose binding protein tolerant to low pH. AB - We have studied the uptake of maltose in the thermoacidophilic gram-positive bacterium Alicyclobacillus acidocaldarius, which grows best at 57 degrees C and pH 3.5. Under these conditions, accumulation of [(14)C]maltose was observed in cells grown with maltose but not in those grown with glucose. At lower temperatures or higher pH values, the transport rates substantially decreased. Uptake of radiolabeled maltose was inhibited by maltotetraose, acarbose, and cyclodextrins but not by lactose, sucrose, or trehalose. The kinetic parameters (K(m) of 0.91 +/- 0.06 microM and V(max) ranging from 0.6 to 3.7 nmol/min/mg of protein) are consistent with a binding protein-dependent ATP binding cassette (ABC) transporter. A corresponding binding protein (MalE) that interacts with maltose with high affinity (K(d) of 1.5 microM) was purified from the culture supernatant of maltose-grown cells. Immunoelectron microscopy revealed distribution of the protein throughout the cell wall. The malE gene was cloned and sequenced. Five additional open reading frames, encoding components of a maltose transport system (MalF and MalG), a putative transcriptional regulator (MalR), a cyclodextrinase (CdaA), and an alpha-glucosidase (GlcA), were identified downstream of malE. The malE gene lacking the DNA sequence that encodes the signal sequence was expressed in Escherichia coli. The purified wild type and recombinant proteins bind maltose with high affinity over a wide pH range (2.5 to 7) and up to 80 degrees C. Recombinant MalE cross-reacted with an antiserum raised against the wild-type protein, thereby indicating that the latter is the product of the malE gene. The MalE protein might be well suited as a model to study tolerance of proteins to low pH. PMID- 11053373 TI - The bulged nucleotide in the Escherichia coli minimal selenocysteine insertion sequence participates in interaction with SelB: a genetic approach. AB - The UGA codon, which usually acts as a stop codon, can also direct the incorporation into a protein of the amino acid selenocysteine. This UGA decoding process requires a cis-acting mRNA element called the selenocysteine insertion sequence (SECIS), which can form a stem-loop structure. In Escherichia coli, selenocysteine incorporation requires only the 17-nucleotide-long upper stem-loop structure of the fdhF SECIS. This structure carries a bulged nucleotide U at position 17. Here we asked whether the single bulged nucleotide located in the upper stem-loop structure of the E. coli fdhF SECIS is involved in the in vivo interaction with SelB. We used a genetic approach, generating and characterizing selB mutations that suppress mutations of the bulged nucleotide in the SECIS. All the selB suppressor mutations isolated were clustered in a region corresponding to 28 amino acids in the SelB C-terminal subdomain 4b. These selB suppressor mutations were also found to suppress mutations in either the loop or the upper stem of the E. coli SECIS. Thus, the E. coli SECIS upper stem-loop structure can be considered a "single suppressible unit," suggesting that there is some flexibility to the nature of the interaction between this element and SelB. PMID- 11053374 TI - Genetics of swarming motility in Salmonella enterica serovar typhimurium: critical role for lipopolysaccharide. AB - Salmonella enterica serovar Typhimurium can differentiate into hyperflagellated swarmer cells on agar of an appropriate consistency (0.5 to 0.8%), allowing efficient colonization of the growth surface. Flagella are essential for this form of motility. In order to identify genes involved in swarming, we carried out extensive transposon mutagenesis of serovar Typhimurium, screening for those that had functional flagella yet were unable to swarm. A majority of these mutants were defective in lipopolysaccharide (LPS) synthesis, a large number were defective in chemotaxis, and some had defects in putative two-component signaling components. While the latter two classes were defective in swarmer cell differentiation, representative LPS mutants were not and could be rescued for swarming by external addition of a biosurfactant. A mutation in waaG (LPS core modification) secreted copious amounts of slime and showed a precocious swarming phenotype. We suggest that the O antigen improves surface "wettability" required for swarm colony expansion, that the LPS core could play a role in slime generation, and that multiple two-component systems cooperate to promote swarmer cell differentiation. The failure to identify specific swarming signals such as amino acids, pH changes, oxygen, iron starvation, increased viscosity, flagellar rotation, or autoinducers leads us to consider a model in which the external slime is itself both the signal and the milieu for swarming motility. The model explains the cell density dependence of the swarming phenomenon. PMID- 11053375 TI - Comparative genetic analysis of Mycobacterium ulcerans and Mycobacterium marinum reveals evidence of recent divergence. AB - Previous studies of the 16S rRNA genes from Mycobacterium ulcerans and Mycobacterium marinum have suggested a very close genetic relationship between these species (99.6% identity). However, these organisms are phenotypically distinct and cause diseases with very different pathologies. To investigate this apparent paradox, we compared 3,306 nucleotides from the partial sequences of eight housekeeping and structural genes derived from 18 M. ulcerans strains and 22 M. marinum strains. This analysis confirmed the close genetic relationship inferred from the 16S rRNA data, with nucleotide sequence identity ranging from 98.1 to 99.7%. The multilocus sequence analysis also confirmed previous genotype studies of M. ulcerans that have identified distinct genotypes within a geographical region. Single isolates of both M. ulcerans and M. marinum that were shown by the sequence analysis to be the most closely related were then selected for further study. One- and two-dimensional pulsed-field gel electrophoresis was employed to compare the architecture and size of the genome from each species. Genome sizes of approximately 4.4 and 4.6 Mb were obtained for M. ulcerans and M. marinum, respectively. Significant macrorestriction fragment polymorphism was observed between the species. However, hybridization analysis of DNA cleaved with more frequently cutting enzymes identified significant preservation of the flanking sequence at seven of the eight loci sequenced. The exception was the 16S rRNA locus. Two high-copy-number insertion sequences, IS2404 and IS2606, have recently been reported in M. ulcerans, and significantly, these elements are not present in M. marinum. Hybridization of the AseI restriction fragments from M. ulcerans with IS2404 and IS2606 indicated widespread genome distribution for both of these repeated sequences. Taken together, these data strongly suggest that M. ulcerans has recently diverged from M. marinum by the acquisition and concomitant loss of DNA in a manner analogous to the emergence of M. tuberculosis, where species diversity is being driven mainly by the activity of mobile DNA elements. PMID- 11053376 TI - A new thermoactive pullulanase from Desulfurococcus mucosus: cloning, sequencing, purification, and characterization of the recombinant enzyme after expression in Bacillus subtilis. AB - The gene encoding a thermoactive pullulanase from the hyperthermophilic anaerobic archaeon Desulfurococcus mucosus (apuA) was cloned in Escherichia coli and sequenced. apuA from D. mucosus showed 45.4% pairwise amino acid identity with the pullulanase from Thermococcus aggregans and contained the four regions conserved among all amylolytic enzymes. apuA encodes a protein of 686 amino acids with a 28-residue signal peptide and has a predicted mass of 74 kDa after signal cleavage. The apuA gene was then expressed in Bacillus subtilis and secreted into the culture fluid. This is one of the first reports on the successful expression and purification of an archaeal amylopullulanase in a Bacillus strain. The purified recombinant enzyme (rapuDm) is composed of two subunits, each having an estimated molecular mass of 66 kDa. Optimal activity was measured at 85 degrees C within a broad pH range from 3.5 to 8.5, with an optimum at pH 5.0. Divalent cations have no influence on the stability or activity of the enzyme. RapuDm was stable at 80 degrees C for 4 h and exhibited a half-life of 50 min at 85 degrees C. By high-pressure liquid chromatography analysis it was observed that rapuDm hydrolyzed alpha-1,6 glycosidic linkages of pullulan, producing maltotriose, and also alpha-1,4 glycosidic linkages in starch, amylose, amylopectin, and cyclodextrins, with maltotriose and maltose as the main products. Since the thermoactive pullulanases known so far from Archaea are not active on cyclodextrins and are in fact inhibited by these cyclic oligosaccharides, the enzyme from D. mucosus should be considered an archaeal pullulanase type II with a wider substrate specificity. PMID- 11053377 TI - BenR, a XylS homologue, regulates three different pathways of aromatic acid degradation in Pseudomonas putida. AB - Pseudomonas putida converts benzoate to catechol using two enzymes that are encoded on the chromosome and whose expression is induced by benzoate. Benzoate also binds to the regulator XylS to induce expression of the TOL (toluene degradation) plasmid-encoded meta pathway operon for benzoate and methylbenzoate degradation. Finally, benzoate represses the ability of P. putida to transport 4 hydroxybenzoate (4-HBA) by preventing transcription of pcaK, the gene encoding the 4-HBA permease. Here we identified a gene, benR, as a regulator of benzoate, methylbenzoate, and 4-HBA degradation genes. A benR mutant isolated by random transposon mutagenesis was unable to grow on benzoate. The deduced amino acid sequence of BenR showed high similarity (62% identity) to the sequence of XylS, a member of the AraC family of regulators. An additional seven genes located adjacent to benR were inferred to be involved in benzoate degradation based on their deduced amino acid sequences. The benABC genes likely encode benzoate dioxygenase, and benD likely encodes 2-hydro-1,2-dihydroxybenzoate dehydrogenase. benK and benF were assigned functions as a benzoate permease and porin, respectively. The possible function of a final gene, benE, is not known. benR activated expression of a benA-lacZ reporter fusion in response to benzoate. It also activated expression of a meta cleavage operon promoter-lacZ fusion inserted in an E. coli chromosome. Third, benR was required for benzoate-mediated repression of pcaK-lacZ fusion expression. The benA promoter region contains a direct repeat sequence that matches the XylS binding site previously defined for the meta cleavage operon promoter. It is likely that BenR binds to the promoter region of chromosomal benzoate degradation genes and plasmid-encoded methylbenzoate degradation genes to activate gene expression in response to benzoate. The action of BenR in repressing 4-HBA uptake is probably indirect. PMID- 11053378 TI - Silencing and activation of ClyA cytotoxin expression in Escherichia coli. AB - Cytolysin A (ClyA) is a pore-forming cytotoxic protein encoded by the clyA gene of Escherichia coli K-12. Genetic analysis suggested that clyA is silenced by the nucleoid protein H-NS. Purified H-NS protein showed preferential binding to clyA sequences in the promoter region, as evidenced by DNase I footprinting and gel mobility shift assays. Transcriptional derepression and activation of a chromosomal clyA::luxAB operon fusion were seen under conditions of H-NS deficiency and SlyA overproduction, respectively. In H-NS-deficient bacteria neither the absence nor the overproduction of SlyA affected the derepressed ClyA expression any further. Therefore, we suggest that overproduction of SlyA in hns(+) E. coli derepresses clyA transcription by counteracting H-NS. The cyclic AMP receptor protein (CRP) was required for ClyA expression, and it interacted with a predicted, albeit suboptimal, CRP binding site in the clyA upstream region. Site-specific alterations of the CRP binding site to match the consensus resulted in substantially higher levels of ClyA expression, while alterations that were predicted to reduce CRP binding reduced ClyA expression. During anaerobic growth the fumarate and nitrate reduction regulator (FNR) was important for ClyA expression, and the clyA gene could be activated by overexpression of FNR. A major clyA transcript having its 5' end (+1) located 72 bp upstream of the translational start codon and 61 bp downstream of the CRP-FNR binding site was detected in the absence of H-NS. The clyA promoter was characterized as a class I promoter that could be transcriptionally activated by CRP and/or FNR. According to DNA bending analyses, the clyA promoter region has high intrinsic curvature. We suggest that it represents a regulatory region which is particularly susceptible to H-NS silencing, and its features are discussed in relation to regulation of other silenced operons. PMID- 11053379 TI - Detection of mRNA transcripts and active transcription in persistent Mycobacterium tuberculosis induced by exposure to rifampin or pyrazinamide. AB - Mycobacterium tuberculosis can persist in an altered physiological state for many years after initial infection, and it may reactivate to cause active disease. An analogous persistent state, possibly consisting of several different subpopulations of bacteria, may arise during chemotherapy; this state is thought to be responsible for the prolonged period required for effective chemotherapy. Using two models of drug-induced persistence, we show that both microaerophilic stationary-phase M. tuberculosis treated with a high dose of rifampin in vitro and pyrazinamide-induced persistent bacteria in mice are nonculturable yet still contain 16S rRNA and mRNA transcripts. Also, the in vitro persistent, plate culture-negative bacteria incorporate radioactive uridine into their RNA in the presence of rifampin and can rapidly up-regulate gene transcription after the replacement of the drug with fresh medium and in response to heat shock. Our results show that persistent M. tuberculosis has transcriptional activity. This finding provides a molecular basis for the rational design of drugs targeted at persistent bacteria. PMID- 11053380 TI - Role of the carboxy terminus of Escherichia coli FtsA in self-interaction and cell division. AB - The role of the carboxy terminus of the Escherichia coli cell division protein FtsA in bacterial division has been studied by making a series of short sequential deletions spanning from residue 394 to 420. Deletions as short as 5 residues destroy the biological function of the protein. Residue W415 is essential for the localization of the protein into septal rings. Overexpression of the ftsA alleles harboring these deletions caused a coiled cell phenotype previously described for another carboxy-terminal mutation (Gayda et al., J. Bacteriol. 174:5362-5370, 1992), suggesting that an interaction of FtsA with itself might play a role in its function. The existence of such an interaction was demonstrated using the yeast two-hybrid system and a protein overlay assay. Even these short deletions are sufficient for impairing the interaction of the truncated FtsA forms with the wild-type protein in the yeast two-hybrid system. The existence of additional interactions between FtsA molecules, involving other domains, can be postulated from the interaction properties shown by the FtsA deletion mutant forms, because although unable to interact with the wild-type and with FtsADelta1, they can interact with themselves and cross-interact with each other. The secondary structures of an extensive deletion, FtsADelta27, and the wild-type protein are indistinguishable when analyzed by Fourier transform infrared spectroscopy, and moreover, FtsADelta27 retains the ability to bind ATP. These results indicate that deletion of the carboxy-terminal 27 residues does not alter substantially the structure of the protein and suggest that the loss of biological function of the carboxy-terminal deletion mutants might be related to the modification of their interacting properties. PMID- 11053381 TI - Complementary metal ion specificity of the metal-citrate transporters CitM and CitH of Bacillus subtilis. AB - Citrate uptake in Bacillus subtilis is stimulated by a wide range of divalent metal ions. The metal ions were separated into two groups based on the expression pattern of the uptake system. The two groups correlated with the metal ion specificity of two homologous B. subtilis secondary citrate transporters, CitM and CitH, upon expression in Escherichia coli. CitM transported citrate in complex with Mg(2+), Ni(2+), Mn(2+), Co(2+), and Zn(2+) but not in complex with Ca(2+), Ba(2+), and Sr(2+). CitH transported citrate in complex with Ca(2+), Ba(2+), and Sr(2+) but not in complex with Mg(2+), Ni(2+), Mn(2+), Co(2+), and Zn(2+). Both transporters did not transport free citrate. Nevertheless, free citrate uptake could be demonstrated in B. subtilis, indicating the expression of at least a third citrate transporter, whose identity is not known. For both the CitM and CitH transporters it was demonstrated that the metal ion promoted citrate uptake and, vice versa, that citrate promoted uptake of the metal ion, indicating that the complex is the transported species. The results indicate that CitM and CitH are secondary transporters that transport complexes of divalent metal ions and citrate but with a complementary metal ion specificity. The potential physiological function of the two transporters is discussed. PMID- 11053382 TI - The malonate decarboxylase operon of Acinetobacter calcoaceticus KCCM 40902 is regulated by malonate and the transcriptional repressor MdcY. AB - A regulatory gene-like open reading frame oriented oppositely to mdcL, coined mdcY, was found upstream from the structural genes of the mdcLMACDEGBH operon in Acinetobacter calcoaceticus KCCM 40902. To elucidate the function of this gene, mdcY was expressed in Escherichia coli, and the MdcY protein was purified to homogeneity. Its DNA binding activity and binding site were examined by gel retardation and footprinting assays in vitro and by site-directed mutagenesis of the binding sites in vivo. The regulator bound target DNA regardless of the presence of malonate, and the binding site was found centered at -65 relative to the mdcL transcriptional start site and contains a 12-bp palindromic structure (5'-ATTGTA/TACAAT-3'). Using a promoter fusion to the reporter gene luc, we found that the promoter P(mdcY) is negatively regulated by MdcY independent of malonate. However, the promoter P(mdcL) recovered its activity in the presence of malonate. When mdcY was introduced into A. calcoaceticus KCCM 40902 in which the gene is inactivated by an IS3 family element, malonate decarboxylase was significantly repressed in cultures growing in acetate, succinate, or Luria Bertani medium. However, in cells growing in malonate, malonate decarboxylase was induced, indicating that MdcY is a transcriptional repressor and that malonate or a product resulting from malonate metabolism should be the intracellular inducer of the mdc operon. PMID- 11053383 TI - H-NS controls pap and daa fimbrial transcription in Escherichia coli in response to multiple environmental cues. AB - A comparative study was completed to determine the influence of various environmental stimuli on the transcription of three different fimbrial operons in Escherichia coli and to determine the role of the histone-like protein H-NS in this environmental regulation. The fimbrial operons studied included the pap operon, which encodes pyelonephritis-associated pili (P pili), the daa operon, which encodes F1845 fimbriae, and the fan operon, which encodes K99 fimbriae. Using lacZYA transcriptional fusions within each of the fimbrial operons, we tested temperature, osmolarity, carbon source, rich medium, oxygen levels, pH, amino acids, solid medium, and iron concentration for their effects on fimbrial gene expression. Low temperature, high osmolarity, glucose as a carbon source, and rich medium repressed transcription of all three operons. High iron did not alter transcription of any of the operons tested, whereas the remaining stimuli had effects on individual operons. For the pap and daa operons, introduction of the hns651 mutation relieved the repression, either fully or partially, due to low temperature, glucose as a carbon source, rich medium, and high osmolarity. Taken together, these data indicate that there are common environmental cues that regulate fimbrial transcription in E. coli and that H-NS is an important environmental regulator for fimbrial transcription in response to several stimuli. PMID- 11053384 TI - The Pseudomonas aeruginosa lectins PA-IL and PA-IIL are controlled by quorum sensing and by RpoS. AB - In Pseudomonas aeruginosa, many exoproduct virulence determinants are regulated via a hierarchical quorum-sensing cascade involving the transcriptional regulators LasR and RhlR and their cognate activators, N-(3-oxododecanoyl)-L homoserine lactone (3O-C12-HSL) and N-butanoyl-L-homoserine lactone (C4-HSL). In this paper, we demonstrate that the cytotoxic lectins PA-IL and PA-IIL are regulated via quorum sensing. Using immunoblot analysis, the production of both lectins was found to be directly dependent on the rhl locus while, in a lasR mutant, the onset of lectin synthesis was delayed but not abolished. The PA-IL structural gene, lecA, was cloned and sequenced. Transcript analysis indicated a monocistronic organization with a transcriptional start site 70 bp upstream of the lecA translational start codon. A lux box-type element together with RpoS (sigma(S)) consensus sequences was identified upstream of the putative promoter region. In Escherichia coli, expression of a lecA::lux reporter fusion was activated by RhlR/C4-HSL, but not by LasR/3O-C12-HSL, confirming direct regulation by RhlR/C4-HSL. Similarly, in P. aeruginosa PAO1, the expression of a chromosomal lecA::lux fusion was enhanced but not advanced by the addition of exogenous C4-HSL but not 3O-C12-HSL. Furthermore, mutation of rpoS abolished lectin synthesis in P. aeruginosa, demonstrating that both RpoS and RhlR/C4-HSL are required. Although the C4-HSL-dependent expression of the lecA::lux reporter in E. coli could be inhibited by the presence of 3O-C12-HSL, this did not occur in P. aeruginosa. This suggests that, in the homologous genetic background, 3O C12-HSL does not function as a posttranslational regulator of the RhlR/C4-HSL dependent activation of lecA expression. PMID- 11053385 TI - Spore photoproduct (SP) lyase from Bacillus subtilis specifically binds to and cleaves SP (5-thyminyl-5,6-dihydrothymine) but not cyclobutane pyrimidine dimers in UV-irradiated DNA. AB - The predominant photolesion in the DNA of UV-irradiated dormant bacterial spores is the thymine dimer 5-thyminyl-5,6-dihydrothymine, commonly referred to as spore photoproduct (SP). A major determinant of SP repair during spore germination is its direct reversal by the enzyme SP lyase, encoded by the splB gene in Bacillus subtilis. SplB protein containing an N-terminal tag of six histidine residues [(6His)SplB] was purified from dormant B. subtilis spores and shown to efficiently cleave SP but not cyclobutane cis,syn thymine-thymine dimers in vitro. In contrast, SplB protein containing an N-terminal 10-histidine tag [(10His)SplB] purified from an Escherichia coli overexpression system was incompetent to cleave SP unless the 10-His tag was first removed by proteolysis at an engineered factor Xa site. To assay the parameters of binding of SplB protein to UV-damaged DNA, a 35-bp double-stranded oligonucleotide was constructed which carried a single pair of adjacent thymines on one strand. Irradiation of the oligonucleotide in aqueous solution or at 10% relative humidity resulted in formation of cyclobutane pyrimidine dimers (Py lozengePy) or SP, respectively. (10His)SplB was assayed for oligonucleotide binding using a DNase I protection assay. In the presence of (10His)SplB, the SP-containing oligonucleotide was selectively protected from DNase I digestion (half-life, >60 min), while the Py lozengePy-containing oligonucleotide and the unirradiated oligonucleotide were rapidly digested by DNase I (half-lives, 6 and 9 min, respectively). DNase I footprinting of (10His)SplB bound to the artificial substrate was carried out utilizing the (32)P end-labeled 35-bp oligonucleotide containing SP. DNase I footprinting showed that SplB protected at least a 9-bp region surrounding SP from digestion with DNase I with the exception of two DNase I-hypersensitive sites within the protected region. (10His)SplB also caused significant enhancement of DNase I digestion of the SP-containing oligonucleotide for at least a full helical turn 3' to the protected region. The data suggest that binding of SP lyase to SP causes significant bending or distortion of the DNA helix in the vicinity of the lesion. PMID- 11053386 TI - The yeast model for batten disease: mutations in BTN1, BTN2, and HSP30 alter pH homeostasis. AB - The BTN1 gene product of the yeast Saccharomyces cerevisiae is 39% identical and 59% similar to human CLN3, which is associated with the neurodegenerative disorder Batten disease. Furthermore, btn1-Delta strains have an elevated activity of the plasma membrane H(+)-ATPase due to an abnormally high vacuolar acidity during the early phase of growth. Previously, DNA microarray analysis revealed that btn1-Delta strains compensate for the altered plasma membrane H(+) ATPase activity and vacuolar pH by elevating the expression of the two genes HSP30 and BTN2. We now show that deletion of either HSP30 or BTN2 in either BTN1(+) or btn1-Delta strains does not alter vacuolar pH but does lead to an increased activity of the vacuolar H(+)-ATPase. Deletion of BTN1, BTN2, or HSP30 does not alter cytosolic pH but diminishes pH buffering capacity and causes poor growth at low pH in a medium containing sorbic acid, a condition known to result in disturbed intracellular pH homeostasis. Btn2p was localized to the cytosol, suggesting a role in mediating pH homeostasis between the vacuole and plasma membrane H(+)-ATPase. Increased expression of HSP30 and BTN2 in btn1-Delta strains and diminished growth of btn1-Delta, hsp30-Delta, and btn2-Delta strains at low pH reinforce our view that altered pH homeostasis is the underlying cause of Batten disease. PMID- 11053387 TI - A DNA ligase from a hyperthermophilic archaeon with unique cofactor specificity. AB - A gene encoding DNA ligase (lig(Tk)) from a hyperthermophilic archaeon, Thermococcus kodakaraensis KOD1, has been cloned and sequenced, and its protein product has been characterized. lig(Tk) consists of 1,686 bp, corresponding to a polypeptide of 562 amino acids with a predicted molecular mass of 64,079 Da. Sequence comparison with previously reported DNA ligases and the presence of conserved motifs suggested that Lig(Tk) was an ATP-dependent DNA ligase. Phylogenetic analysis indicated that Lig(Tk) was closely related to the ATP dependent DNA ligase from Methanobacterium thermoautotrophicum DeltaH, a moderate thermophilic archaeon, along with putative DNA ligases from Euryarchaeota and Crenarchaeota. We expressed lig(Tk) in Escherichia coli and purified the recombinant protein. Recombinant Lig(Tk) was monomeric, as is the case for other DNA ligases. The protein displayed DNA ligase activity in the presence of ATP and Mg(2+). The optimum pH of Lig(Tk) was 8.0, the optimum concentration of Mg(2+), which was indispensable for the enzyme activity, was 14 to 18 mM, and the optimum concentration of K(+) was 10 to 30 mM. Lig(Tk) did not display single-stranded DNA ligase activity. At enzyme concentrations of 200 nM, we observed significant DNA ligase activity even at 100 degrees C. Unexpectedly, Lig(Tk) displayed a relatively small, but significant, DNA ligase activity when NAD(+) was added as the cofactor. Treatment of NAD(+) with hexokinase did not affect this activity, excluding the possibility of contaminant ATP in the NAD(+) solution. This unique cofactor specificity was also supported by the observation of adenylation of Lig(Tk) with NAD(+). This is the first biochemical study of a DNA ligase from a hyperthermophilic archaeon. PMID- 11053388 TI - Heterologous NNR-mediated nitric oxide signaling in Escherichia coli. AB - The transcription factor NNR from Paracoccus denitrificans was expressed in a strain of Escherichia coli carrying a plasmid-borne fusion of the melR promoter to lacZ, with a consensus FNR-binding site 41.5 bp upstream of the transcription start site. This promoter was activated by NNR under anaerobic growth conditions in media containing nitrate, nitrite, or the NO(+) donor sodium nitroprusside. Activation by nitrate was abolished by a mutation in the molybdenum cofactor biosynthesis pathway, indicating a requirement for nitrate reductase activity. Activation by nitrate was modulated by the inclusion of reduced hemoglobin in culture media, because of the ability of hemoglobin to sequester nitric oxide and nitrite. The ability of nitrate and nitrite to activate NNR is likely due to the formation of NO (or related species) during nitrate and nitrite respiration. Amino acids potentially involved in NNR activity were replaced by site-directed mutagenesis, and the activities of NNR derivatives were tested in the E. coli reporter system. Substitutions at Cys-103 and Tyr-35 significantly reduced NNR activity but did not abolish the response to reactive nitrogen species. Substitutions at Phe-82 and Tyr-93 severely impaired NNR activity, but the altered proteins retained the ability to repress an FNR-repressible promoter, so these mutations have a "positive control" phenotype. It is suggested that Phe-82 and Tyr-93 identify an activating region of NNR that is involved in an interaction with RNA polymerase. Replacement of Ser-96 with alanine abolished NNR activity, and the protein was undetectable in cell extracts. In contrast, NNR in which Ser-96 was replaced with threonine retained full activity. PMID- 11053389 TI - Characterization of the collagen-binding S-layer protein CbsA of Lactobacillus crispatus. AB - The cbsA gene of Lactobacillus crispatus strain JCM 5810, encoding a protein that mediates adhesiveness to collagens, was characterized and expressed in Escherichia coli. The cbsA open reading frame encoded a signal sequence of 30 amino acids and a mature polypeptide of 410 amino acids with typical features of a bacterial S-layer protein. The cbsA gene product was expressed as a His tag fusion protein, purified by affinity chromatography, and shown to bind solubilized as well as immobilized type I and IV collagens. Three other Lactobacillus S-layer proteins, SlpA, CbsB, and SlpnB, bound collagens only weakly, and sequence comparisons of CbsA with these S-layer proteins were used to select sites in cbsA where deletions and mutations were introduced. In addition, hybrid S-layer proteins that contained the N or the C terminus from CbsA, SlpA, or SlpnB as well as N- and C-terminally truncated peptides from CbsA were constructed by gene fusion. Analysis of these molecules revealed the major collagen-binding region within the N-terminal 287 residues and a weaker type I collagen-binding region in the C terminus of the CbsA molecule. The mutated or hybrid CbsA molecules and peptides that failed to polymerize into a periodic S layer did not bind collagens, suggesting that the crystal structure with a regular array is optimal for expression of collagen binding by CbsA. Strain JCM 5810 was found to contain another S-layer gene termed cbsB that was 44% identical in sequence to cbsA. RNA analysis showed that cbsA, but not cbsB, was transcribed under laboratory conditions. S-layer-protein-expressing cells of strain JCM 5810 adhered to collagen-containing regions in the chicken colon, suggesting that CbsA mediated collagen binding represents a true tissue adherence property of L. crispatus. PMID- 11053390 TI - Novel toluene elimination system in a toluene-tolerant microorganism. AB - In studies of Pseudomonas putida IH-2000, a toluene-tolerant microorganism, membrane vesicles (MVs) were found to be released from the outer membrane when toluene was added to the culture. These MVs were found to be composed of phospholipids, lipopolysaccharides (LPS), and very low amounts of outer membrane proteins. The MVs also contained a higher concentration of toluene molecules (0.172 +/- 0. 012 mol/mol of lipid) than that found in the cell membrane. In contrast to the wild-type strain, the toluene-sensitive mutant strain 32, which differs from the parent strain in LPS and outer membrane proteins, did not release MVs from the outer membrane. The toluene molecules adhering to the outer membrane are eliminated by the shedding of MVs, and this system appears to serve as an important part of the toluene tolerance system of IH-2000. PMID- 11053391 TI - Characterization of a novel outer membrane hemin-binding protein of Porphyromonas gingivalis. AB - Porphyromonas gingivalis is a gram-negative, anaerobic coccobacillus that has been implicated as a major etiological agent in the development of chronic periodontitis. In this paper, we report the characterization of a protein, IhtB (iron heme transport; formerly designated Pga30), that is an outer membrane hemin binding protein potentially involved in iron assimilation by P. gingivalis. IhtB was localized to the cell surface of P. gingivalis by Western blot analysis of a Sarkosyl-insoluble outer membrane preparation and by immunocytochemical staining of whole cells using IhtB peptide-specific antisera. The protein, released from the cell surface, was shown to bind to hemin using hemin-agarose. The growth of heme-limited, but not heme-replete, P. gingivalis cells was inhibited by preincubation with IhtB peptide-specific antisera. The ihtB gene was located between an open reading frame encoding a putative TonB-linked outer membrane receptor and three open reading frames that have sequence similarity to ATP binding cassette transport system operons in other bacteria. Analysis of the deduced amino acid sequence of IhtB showed significant similarity to the Salmonella typhimurium protein CbiK, a cobalt chelatase that is structurally related to the ATP-independent family of ferrochelatases. Molecular modeling indicated that the IhtB amino acid sequence could be threaded onto the CbiK fold with the IhtB structural model containing the active-site residues critical for chelatase activity. These results suggest that IhtB is a peripheral outer membrane chelatase that may remove iron from heme prior to uptake by P. gingivalis. PMID- 11053392 TI - Bacillus subtilis SMC is required for proper arrangement of the chromosome and for efficient segregation of replication termini but not for bipolar movement of newly duplicated origin regions. AB - SMC protein is required for chromosome condensation and for the faithful segregation of daughter chromosomes in Bacillus subtilis. The visualization of specific sites on the chromosome showed that newly duplicated origin regions in growing cells of an smc mutant were able to segregate from each other but that the location of origin regions was frequently aberrant. In contrast, the segregation of replication termini was impaired in smc mutant cells. This analysis was extended to germinating spores of an smc mutant. The results showed that during germination, newly duplicated origins, but not termini, were able to separate from each other in the absence of SMC. Also, DAPI (4',6'-diamidino-2 phenylindole) staining revealed that chromosomes in germinating spores were able to undergo partial or complete replication but that the daughter chromosomes were blocked at a late stage in the segregation process. These findings were confirmed by time-lapse microscopy, which showed that after duplication in growing cells the origin regions underwent rapid movement toward opposite poles of the cell in the absence of SMC. This indicates that SMC is not a required component of the mitotic motor that initially drives origins apart after their duplication. It is also concluded that SMC is needed to maintain the proper layout of the chromosome in the cell and that it functions in the cell cycle after origin separation but prior to complete segregation or replication of daughter chromosomes. It is proposed here that chromosome segregation takes place in at least two steps: an SMC-independent step in which origins move apart and a subsequent SMC-dependent step in which newly duplicated chromosomes condense and are thereby drawn apart. PMID- 11053393 TI - Sequence of the genome of Salmonella bacteriophage P22. AB - The sequence of the nonredundant region of the Salmonella enterica serovar Typhimurium temperate, serotype-converting bacteriophage P22 has been completed. The genome is 41,724 bp with an overall moles percent GC content of 47.1%. Numerous examples of potential integration host factor and C1-binding sites were identified in the sequence. In addition, five potential rho-independent terminators were discovered. Sixty-five genes were identified and annotated. While many of these had been described previously, we have added several new ones, including the genes involved in serotype conversion and late control. Two of the serotype conversion gene products show considerable sequence relatedness to GtrA and -B from Shigella phages SfII, SfV, and SfX. We have cloned the serotype-converting cassette (gtrABC) and demonstrated that it results in Salmonella serovar Typhimurium LT2 cells which express antigen O1. Many of the putative proteins show sequence relatedness to proteins from a great variety of other phages, supporting the hypothesis that this phage has evolved through the recombinational exchange of genetic information with other viruses. PMID- 11053394 TI - Development and dynamics of Pseudomonas sp. biofilms. AB - Pseudomonas sp. strain B13 and Pseudomonas putida OUS82 were genetically tagged with the green fluorescent protein and the Discosoma sp. red fluorescent protein, and the development and dynamics occurring in flow chamber-grown two-colored monospecies or mixed-species biofilms were investigated by the use of confocal scanning laser microscopy. Separate red or green fluorescent microcolonies were formed initially, suggesting that the initial small microcolonies were formed simply by growth of substratum attached cells and not by cell aggregation. Red fluorescent microcolonies containing a few green fluorescent cells and green fluorescent microcolonies containing a few red fluorescent cells were frequently observed in both monospecies and two-species biofilms, suggesting that the bacteria moved between the microcolonies. Rapid movement of P. putida OUS82 bacteria inside microcolonies was observed before a transition from compact microcolonies to loose irregularly shaped protruding structures occurred. Experiments involving a nonflagellated P. putida OUS82 mutant suggested that the movements between and inside microcolonies were flagellum driven. The results are discussed in relation to the prevailing hypothesis that biofilm bacteria are in a physiological state different from planktonic bacteria. PMID- 11053395 TI - Characterization of SepL of enterohemorrhagic Escherichia coli. AB - The sepL gene is expressed in the locus of enterocyte effacement and therefore is most likely implicated in the attaching and effacing process, as are the products encoded by open reading frames located up- and downstream of this gene. In this study, the sepL gene of the enterohemorrhagic Escherichia coli (EHEC) strain EDL933 was analyzed and the corresponding polypeptide was characterized. We found that sepL is transcribed monocistronically and independently from the esp operon located downstream, which codes for the secreted proteins EspA, -D, and -B. Primer extension analysis allowed us to identify a single start of transcription 83 bp upstream of the sepL start codon. The analysis of the upstream regions led to the identification of canonical promoter sequences between positions -5 and 36. Translational fusions using lacZ as a reporter gene demonstrated that sepL is activated in the exponential growth phase by stimuli that are characteristic for the intestinal niche, e.g., a temperature of 37 degrees C, a nutrient-rich environment, high osmolarity, and the presence of Mn(2+). Protein localization studies showed that SepL was present in the cytoplasm and associated with the bacterial membrane fraction. To analyze the functional role of the SepL protein during infection of eukaryotic cells, an in-frame deletion mutant was generated. This sepL mutant was strongly impaired in its ability to attach to HeLa cells and induce a local accumulation of actin. These defects were partially restored by providing the sepL gene in trans. The EDL933DeltasepL mutant also exhibited an impaired secretion but not biosynthesis of Esp proteins, which was fully complemented by providing sepL in trans. These results demonstrate the crucial role played by SepL in the biological cycle of EHEC. PMID- 11053396 TI - Evolutionary conservation of methyl-accepting chemotaxis protein location in Bacteria and Archaea. AB - The methyl-accepting chemotaxis proteins (MCPs) are concentrated at the cell poles in an evolutionarily diverse panel of bacteria and an archeon. In elongated cells, the MCPs are located both at the poles and at regions along the length of the cells. Together, these results suggest that MCP location is evolutionarily conserved. PMID- 11053397 TI - Transcription initiation-defective forms of sigma(54) that differ in ability To function with a heteroduplex DNA template. AB - Transcription by sigma(54)-RNA polymerase holoenzyme requires an activator that catalyzes isomerization of the closed promoter complex to an open complex. We examined mutant forms of Salmonella enterica serovar Typhimurium sigma(54) that were defective in transcription initiation but retained core RNA polymerase- and promoter-binding activities. Four of the mutant proteins allowed activator independent transcription from a heteroduplex DNA template. One of these mutant proteins, L124P V148A, had substitutions in a sequence that had not been shown previously to participate in the prevention of activator-independent transcription. The remaining mutants did not allow efficient activator independent transcription from the heteroduplex DNA template and had substitutions within a conserved 20-amino-acid segment (Leu-179 to Leu-199), suggesting a role for this sequence in transcription initiation. PMID- 11053398 TI - Escherichia coli TehB requires S-adenosylmethionine as a cofactor to mediate tellurite resistance. AB - The Escherichia coli chromosomal determinant for tellurite resistance consists of two genes (tehA and tehB) which, when expressed on a multicopy plasmid, confer resistance to K(2)TeO(3) at 128 microg/ml, compared to the MIC of 2 microg/ml for the wild type. TehB is a cytoplasmic protein which possesses three conserved motifs (I, II, and III) found in S-adenosyl-L-methionine (SAM)-dependent non nucleic acid methyltransferases. Replacement of the conserved aspartate residue in motif I by asparagine or alanine, or of the conserved phenylalanine in motif II by tyrosine or alanine, decreased resistance to background levels. Our results are consistent with motifs I and II in TehB being involved in SAM binding. Additionally, conformational changes in TehB are observed upon binding of both tellurite and SAM. The hydrodynamic radius of TehB measured by dynamic light scattering showed a approximately 20% decrease upon binding of both tellurite and SAM. These data suggest that TehB utilizes a methyltransferase activity in the detoxification of tellurite. PMID- 11053399 TI - Second-site suppressor mutations of inactivating substitutions at gly247 of the tetracycline efflux protein, Tet(B). AB - An Asp or Asn substitution for Gly247 in transmembrane helix 8 (TM-8) of Tet(B), the tetracycline efflux protein, eliminated tetracycline resistance. Second site suppressor mutations which partially restored resistance were located in TM-5, 8, -10, or -11 or in cytoplasmic loop 8-9 or loop 10-11. These results indicate physical proximity or functional relationships between TM-8 and these other regions of Tet(B). PMID- 11053400 TI - Exfoliatin-producing strains define a fourth agr specificity group in Staphylococcus aureus. AB - The staphylococcal virulon is activated by the density-sensing agr system, which is autoinduced by a short peptide (autoinducing peptide [AIP]) processed from a propeptide encoded by agrD. A central segment of the agr locus, consisting of the C-terminal two-thirds of AgrB (the putative processing enzyme), AgrD, and the N terminal half of AgrC (the receptor), shows striking interstrain variation. This finding has led to the division of Staphylococcus aureus isolates into three different agr specificity groups and to the division of non-aureus staphylococci into a number of others. The AIPs cross-inhibit the agr responses between groups. We have previously shown that most menstrual toxic shock strains belong to agr specificity group III but that no strong clinical identity has been associated with strains of the other two groups. In the present report, we demonstrate a fourth agr specificity group among S. aureus strains and show that most exfoliatin-producing strains belong to this group. A striking common feature of group IV strains is activation of the agr response early in exponential phase, at least 2 h earlier than in strains of the other groups. This finding raises the question of the biological significance of the agr autoinduction threshold. PMID- 11053401 TI - A gene of Synechocystis sp. Strain PCC 6803 encoding a novel iron transporter. AB - A mutant of Synechocystis sp. strain PCC 6803 disrupted for sll1878 exhibited greatly reduced Fe(3+) transport activity. The K(m) value of sll1878-dependent Fe(3+) transport in cells grown in iron-replete medium was 0.5 microM. Both the maximal rate and K(m) value were increased in iron-starved cells. PMID- 11053402 TI - Cholic acid is accumulated spontaneously, driven by membrane deltapH, in many lactobacilli. AB - Many lactobacilli from various origins were found to apparently lack cholic acid extrusion activity. Cholic acid was accumulated spontaneously, driven by the transmembrane proton gradient. Accumulation is a newly identified kind of interaction between intestinal microbes and unconjugated bile acids and is different from extrusion and modification, which have been described previously. PMID- 11053403 TI - Genes for the type IV secretion system in an intracellular symbiont, Wolbachia, a causative agent of various sexual alterations in arthropods. AB - Wolbachia species are intracellular bacteria known to cause reproductive abnormalities in their hosts. In this study, we identified Wolbachia genes encoding homologs to the type IV secretion system by which many pathogenic bacteria secrete macromolecules. The genes identified encoded most of the essential components of the secretion system and were cotranscribed as an operon. PMID- 11053404 TI - Analysis of the role of recA in phenotypic switching of Pseudomonas tolaasii. AB - Switching between the pathogenic smooth (1116S) and nonpathogenic rough (1116R) forms of Pseudomonas tolaasii occurs due to the reversible duplication of a 661 bp element within the pheN locus. Disruption of the chromosomal recA locus of 1116S and 1116R produced strains 1116SrecA and 1116RrecA, respectively, which showed typical loss of UV resistance. Switching from the smooth to the rough form was virtually eliminated in the 1116SrecA strain, whereas the extent of switching from the rough to the smooth form was almost identical in 1116R and 1116RrecA. It is concluded that phenotypic switching from 1116S to 1116R is recA dependent whereas that from 1116R to 1116S is recA independent. PMID- 11053405 TI - Identification of an ancillary protein, YabF, required for activity of the KefC glutathione-gated potassium efflux system in Escherichia coli. AB - A new subunit, YabF, for the KefC K(+) efflux system in Escherichia coli has been identified. The subunit is required for maximum activity of KefC. Deletion of yabF reduces KefC activity 10-fold, and supply of YabF in trans restores activity. IS2 and IS10R insertions in yabF can be isolated as suppressors of KefC activity consequent upon the V427A and D264A KefC mutations. PMID- 11053406 TI - DAX-1 functions as an LXXLL-containing corepressor for activated estrogen receptors. AB - We have discovered that the orphan receptor DAX-1 (NROB1) interacts with the estrogen receptors ERalpha and ERbeta. Interaction occurs with ligand-activated ERs in solution and on DNA and is mediated by the unique DAX-1 N-terminal repeat domain. Each of the three repeats contains a leucine-rich receptor-binding motif, known as the LXXLL motif, which is usually found in nuclear receptor coactivators. We have demonstrated that DAX-1 functions as an inhibitor of ER activation in mammalian cells and suggest a mechanism involving two sequential events, occupation of the ligand-induced coactivator-binding surface and subsequent recruitment of corepressors. Accordingly, we propose that DAX-1 itself acts as a corepressor for ERs. Because DAX-1 is coexpressed with ERs in reproductive tissues, these interactions could play significant roles by influencing estrogen signaling pathways. Our results point at functional similarities between DAX-1 and the orphan receptor SHP (NROB2) in that they have acquired features of transcriptional coregulators that are unique for members of the nuclear receptor family. PMID- 11053407 TI - The Lys1010-Lys1325 fragment of the Wilson's disease protein binds nucleotides and interacts with the N-terminal domain of this protein in a copper-dependent manner. AB - Wilson's disease, an autosomal disorder associated with vast accumulation of copper in tissues, is caused by mutations in a gene encoding a copper transporting ATPase (Wilson's disease protein, WNDP). Numerous mutations have been identified throughout the WNDP sequence, particularly in the Lys(1010) Lys(1325) segment; however, the biochemical properties and molecular mechanism of WNDP remain poorly characterized. Here, the Lys(1010)-Lys(1325) fragment of WNDP was overexpressed, purified, and shown to form an independently folded ATP binding domain (ATP-BD). ATP-BD binds the fluorescent ATP analogue trinitrophenyl ATP with high affinity, and ATP competes with trinitrophenyl-ATP for the binding site; ADP and AMP appear to bind to ATP-BD at the site separate from ATP. Purified ATP-BD hydrolyzes ATP and interacts specifically with the N-terminal copper-binding domain of WNDP (N-WNDP). Strikingly, copper binding to N-WNDP diminishes these interactions, suggesting that the copper-dependent change in domain-domain contact may represent the mechanism of WNDP regulation. In agreement with this hypothesis, N-WNDP induces conformational changes in ATP-BD as evidenced by the altered nucleotide binding properties of ATP-BD in the presence of N-WNDP. Significantly, the effects of copper-free and copper-bound N WNDP on ATP-BD are not identical. The implications of these results for the WNDP function are discussed. PMID- 11053408 TI - Coupling of heterotrimeric Gi proteins to the erythropoietin receptor. AB - To identify new proteins involved in erythropoietin (Epo) signal transduction, we purified the entire set of proteins reactive with anti-phosphotyrosine antibodies from Epo-stimulated UT7 cells. Antisera generated against these proteins were used to screen a lambdaEXlox expression library. One of the isolated cDNAs encodes Gbeta2, the beta2 subunit of heterotrimeric GTP-binding proteins. Gbeta and Galpha(i) coprecipitated with the Epo receptor (EpoR) in extracts from human and murine cell lines and from normal human erythroid progenitor cells. In addition, in vitro Gbeta associated with a fusion protein containing the intracellular domain of the EpoR. Using EpoR mutants, we found that the distal part of the EpoR (between amino acids 459-479) was required for Gi binding. Epo activation of these cells induced the release of the Gi protein from the EpoR. Moreover in isolated cell membranes, Epo treatment inhibited ADP-ribosylation of Gi and increased the binding of GTP. Our results show that heterotrimeric Gi proteins associate with the C-terminal end of the EpoR. Receptor activation leads to the activation and dissociation of Gi from the receptor, suggesting a functional role of Gi protein in Epo signal transduction. PMID- 11053409 TI - JAM-2, a novel immunoglobulin superfamily molecule, expressed by endothelial and lymphatic cells. AB - Cell-cell contacts are essential for morphogenesis and tissue function and play a vital role in mediating endothelial cohesion within the vascular system during vessel growth and organization. We identified a novel junctional adhesion molecule, named JAM-2, by a selective RNA display method, which allowed identification of transcripts encoding immunoglobulin superfamily molecules regulated during coculture of endothelial cells with tumor cells. The JAM-2 transcript is highly expressed during embryogenesis and is detected in lymph node and Peyer's patches RNA of adult mice. Accordingly, antibodies specific for JAM-2 stain high endothelial venules and lymphatic vessels in lymphoid organs, and vascular structures in the kidney. Using real time video microscopy, we show that JAM-2 is localized within minutes to the newly formed cell-cell contact. The role of the protein in the sealing of cell-cell contact is further suggested by the reduced paracellular permeability of cell monolayer transfected with JAM-2 cDNA, and by the localization of JAM-2 to tight junctional complexes of polarized cells. Taken together, our results suggest that JAM-2 is a novel vascular molecule, which participates in interendothelial junctional complexes. PMID- 11053410 TI - Reaction of human myoglobin and nitric oxide. Heme iron or protein sulfhydryl (s) nitrosation dependence on the absence or presence of oxygen. AB - The amino acid sequence of human myoglobin (Mb) is similar to other mammalian Mb except for a unique cysteine residue at position 110 (Cys(110)). Anaerobic treatment of ferrous forms of wild-type human Mb, the C110A variant of human Mb or horse heart Mb, with either authentic NO or chemically derived NO in vitro yields heme-NO complexes as detected by electron paramagnetic resonance spectroscopy (EPR). By contrast, no EPR-detectable heme-NO complex was observed from the aerobic reactions of NO and either the ferric or oxy-Mb forms of wild type human or horse heart myoglobins. Mass analyses of wild-type human Mb treated aerobically with NO indicated a mass increase of approximately 30 atomic mass units (i.e., NO/Mb = 1 mol/mol). Mass analyses of the corresponding apoprotein after heme removal showed that NO was associated with the apoprotein fraction. New electronic maxima were detected at A(333 nm) (epsilon = 3665 +/- 90 mol(-)(1) cm(-)(1); mean +/- S.D.) and A(545 nm) (epsilon = 44 +/- 3 mol(-)(1) cm(-)(1)) in solutions of S-nitrosated wild-type human Mb (similar to S-nitrosoglutathione). Importantly, the sulfhydryl S-H stretch vibration for Cys(110) measured by Fourier transform infrared (nu approximately 2552 cm(-)(1)) was absent for both holo- and apo- forms of the wild-type human protein after aerobic treatment of the protein with NO. Together, these data indicate that the reaction of wild-type human Mb and NO yields either heme-NO or a novel S-nitrosated protein dependent on the oxidation state of the heme iron and the presence or absence of dioxygen. PMID- 11053411 TI - Most pathogenic mutations do not alter the membrane topology of the prion protein. AB - The prion protein (PrP), a glycolipid-anchored membrane glycoprotein, contains a conserved hydrophobic sequence that can span the lipid bilayer in either direction, resulting in two transmembrane forms designated (Ntm)PrP and (Ctm)PrP. Previous studies have shown that the proportion of (Ctm)PrP is increased by mutations in the membrane-spanning segment, and it has been hypothesized that (Ctm)PrP represents a key intermediate in the pathway of prion-induced neurodegeneration. To further test this idea, we have surveyed a number of mutations associated with familial prion diseases to determine whether they alter the proportions of (Ntm)PrP and (Ctm)PrP produced in vitro, in transfected cells, and in transgenic mice. For the in vitro experiments, PrP mRNA was translated in the presence of murine thymoma microsomes which, in contrast to the canine pancreatic microsomes used in previous studies, are capable of efficient glycolipidation. We confirmed that mutations within or near the transmembrane domain enhance the formation of (Ctm)PrP, and we demonstrate for the first time that this species contains a C-terminal glycolipid anchor, thus exhibiting an unusual, dual mode of membrane attachment. However, we find that pathogenic mutations in other regions of the molecule have no effect on the amounts of (Ctm)PrP and (Ntm)PrP, arguing against the proposition that transmembrane PrP plays an obligate role in the pathogenesis of prion diseases. PMID- 11053412 TI - The BRN-3A transcription factor protects sensory but not sympathetic neurons from programmed cell death/apoptosis. AB - Inactivation of the gene encoding the POU domain transcription factor BRN-3A results in the absence of specific neurons in knockout mice. Here we demonstrate for the first time a direct effect of BRN-3A on the survival of neuronal cells. Specifically, overexpression of BRN-3A in cultured trigeminal ganglion or dorsal root ganglion sensory neurons enhanced their survival following the withdrawal of nerve growth factor. Moreover, reduction of BRN-3A levels impaired the survival of these neurons. The survival of sympathetic neurons was not affected by either approach. Similarly, overexpression of BRN-3A activated the endogenous Bcl-2 gene in trigeminal neurons, but not in sympathetic neurons. The protective effect of BRN-3A on trigeminal neuron survival following nerve growth factor withdrawal significantly increased during embryonic development. In contrast, overexpression of the related factor BRN-3B enhanced survival of trigeminal neurons only at an early stage of embryonic development. Thus, BRN-3A (and in some circumstances, BRN-3B) can promote the survival of nerve growth factor-dependent sensory but not sympathetic neurons, allowing it to play a direct role in the survival of some (but not all) neuronal populations in the developing and adult nervous systems. PMID- 11053413 TI - Caspase-3-mediated processing of poly(ADP-ribose) glycohydrolase during apoptosis. AB - Poly(ADP-ribose) glycohydrolase (PARG) is responsible for the catabolism of poly(ADP-ribose) synthesized by poly(ADP-ribose) polymerase (PARP-1) and other PARP-1-like enzymes. In this work, we report that PARG is cleaved during etoposide-, staurosporine-, and Fas-induced apoptosis in human cells. This cleavage is concomitant with PARP-1 processing and generates two C-terminal fragments of 85 and 74 kDa. In vitro cleavage assays using apoptotic cell extracts showed that a protease of the caspase family is responsible for PARG processing. A complete inhibition of this cleavage was achieved at nanomolar concentrations of the caspase inhibitor acetyl-Asp-Glu-Val-Asp-aldehyde, suggesting the involvement of caspase-3-like proteases. Consistently, recombinant caspase-3 efficiently cleaved PARG in vitro, suggesting the involvement of this protease in PARG processing in vivo. Furthermore, caspase-3-deficient MCF-7 cells did not show any PARG cleavage in response to staurosporine treatment. The cleavage sites identified by site-directed mutagenesis are DEID(256) downward arrow V and the unconventional site MDVD(307) downward arrow N. Kinetic studies have shown similar maximal velocity (V(max)) and affinity (K(m)) for both full length PARG and its apoptotic fragments, suggesting that caspase-3 may affect PARG function without altering its enzymatic activity. The early cleavage of both PARP-1 and PARG by caspases during apoptosis suggests an important function for poly(ADP-ribose) metabolism regulation during this cell death process. PMID- 11053414 TI - Requirement of homocitrate for the transfer of a 49V-labeled precursor of the iron-vanadium cofactor from VnfX to nif-apodinitrogenase. AB - A vanadium- and iron-containing cluster has been shown previously to accumulate on VnfX in the Azotobacter vinelandii mutant strain CA11.1 (DeltanifHDKvnfDGK::spc). In the present study, we show the homocitrate-dependent transfer of (49)V label from VnfX to nif-apodinitrogenase in vitro. This transfer of radiolabel correlates with acquisition of acetylene reduction activity. Acetylene is reduced both to ethylene and ethane by the hybrid holodinitrogenase so formed, a feature characteristic of alternative nitrogenases. Structural analogues of homocitrate prevent the acetylene reduction ability of the resulting dinitrogenase. Addition of NifB cofactor (-co) or a source of vanadium (Na(3)VO(4) or VCl(3)) does not increase nitrogenase activity. Our results suggest that there is in vitro incorporation of homocitrate into the V-Fe-S cluster associated with VnfX and that the completed cluster can be inserted into nif-apodinitrogenase. The homocitrate incorporation reaction and the insertion of the cluster into nif-apodinitrogenase (alpha(2)beta(2)gamma(2)) do not require MgATP. Attempts to achieve FeV-co synthesis using extracts of other FeV-co negative mutants were unsuccessful, showing that earlier steps in FeV-co synthesis, such as the steps requiring VnfNE or VnfH, do not occur in vitro. PMID- 11053415 TI - Tumor necrosis factor-alpha activation of the c-Jun N-terminal kinase pathway in human neutrophils. Integrin involvement in a pathway leading from cytoplasmic tyrosine kinases apoptosis. AB - The intensity and duration of an inflammatory response depends on the balance of factors that favor perpetuation versus resolution. At sites of inflammation, neutrophils adherent to other cells or matrix components are exposed to tumor necrosis factor-alpha (TNFalpha). Although TNFalpha has been implicated in induction of pro-inflammatory responses, it may also inhibit the intensity of neutrophilic inflammation by promoting apoptosis. Since TNFalpha is not only an important activator of the stress-induced pathways leading to p38 MAPk and c-Jun N-terminal kinase (JNK) but also a potent effector of apoptosis, we investigated the effects of TNFalpha on the JNK pathway in adherent human neutrophils and the potential involvement of this pathway in neutrophil apoptosis. Stimulation with TNFalpha was found to result in beta2 integrin-mediated activation of the cytoplasmic tyrosine kinases Pyk2 and Syk, and activation of a three-part MAPk module composed of MEKK1, MKK7, and/or MKK4 and JNK1. JNK activation was attenuated by blocking antibodies to beta2 integrins, the tyrosine kinase inhibitors, genistein, and tyrphostin A9, a Pyk2-specific inhibitor, and piceatannol, a Syk-specific inhibitor. Exposure of adherent neutrophils to TNFalpha led to the rapid onset of apoptosis that was demonstrated by augmented annexin V binding and caspase-3 cleavage. TNFalpha-induced increases in annexin V binding to neutrophils were attenuated by blocking antibodies to beta2 integrins, and the caspase-3 cleavage was attenuated by tyrphostin A9. Hence, exposure of adherent neutrophils to TNFalpha leads to utilization of the JNK-signaling pathways that may contribute to diverse functional responses including induction of apoptosis and subsequent resolution of the inflammatory response. PMID- 11053416 TI - Camel single-domain antibodies as modular building units in bispecific and bivalent antibody constructs. AB - Single-domain antibodies against various antigens are isolated from the unique heavy-chain antibodies of immunized camels and llamas. These minimal sized binders are very robust and bind the antigen with high affinity in a monomeric state. We evaluated the feasibility to produce soluble, functional bispecific and bivalent antibodies in Escherichia coli with camel single-domain antibody fragments as building blocks. Two single-domain antibody fragments were tethered by the structural upper hinge of a natural antibody to generate bispecific molecules. This linker was chosen for its protease resistance in serum and its natural flexibility to reorient the upstream and downstream located domains. The expression levels, ease of purification, and the solubility of the recombinant proteins were comparable with those of the constituent monomers. The individual moieties fully retain the binding capacity and the binding characteristics within the recombinant bispecific constructs. The easy generation steps and the biophysical properties of these bispecific and bivalent constructs based on camel single-domain antibody fragments makes them particularly attractive for use in therapeutic or diagnostic programs. PMID- 11053417 TI - Domain effects on the DNA-interactive properties of bacteriophage T4 gene 32 protein. AB - Bacteriophage T4 gene 32 protein, a model for single-strand specific nucleic acid binding proteins, consists of three structurally and functionally distinct domains. We have studied the effects of the N and C domains on the protein structure and its nucleic acid-interactive properties. Although the presence of the C domain decreases the proteolytic susceptibility of the core (central) domain, quenching of the core tryptophan fluorescence by iodide is unaltered by the presence of the terminal domains. These results suggest that the overall conformation of the core domain remains largely independent of the flanking domains. Removal of the N or the C terminus does not abolish the DNA renaturation activity of the protein. However, intact protein and its three truncated forms differ in DNA helix-destabilizing activity. The C domain alone is responsible for the kinetic barrier to natural DNA helix destabilization seen with intact protein. Intact protein and core domain potentiate the DNA helix-destabilizing activity of truncated protein lacking only the C domain (*I), enhancing the observed hyperchromicity while increasing the melting temperature. Proteolysis experiments suggest that the affinity of core domain for single-stranded DNA is increased in the presence of *I. We propose that *I can "mingle" with intact protein or core domain while bound to single-stranded DNA. PMID- 11053418 TI - Cell cycle-dependent and DNA damage-inducible nuclear localization of FEN-1 nuclease is consistent with its dual functions in DNA replication and repair. AB - Flap endonuclease-1 (FEN-1), a 43-kDa protein, is a structure-specific and multifunctional nuclease. It plays important roles in RNA primer removal of Okazaki fragments during DNA replication, DNA base excision repair, and maintenance of genome stability. Three functional motifs of the enzyme were proposed to be responsible for its nuclease activities, interaction with proliferating cell nuclear antigen, and nuclear localization. In this study, we demonstrate in HeLa cells that a signal located at the C terminus (the nuclear localization signal (NLS) motif) facilitates nuclear localization of the enzyme during S phase of the cell cycle and in response to DNA damage. Truncation of the NLS motif prevents migration of the protein from the cytoplasm to the nucleus, while having no effect on the nuclease activities and its proliferating cell nuclear antigen interaction capability. Site-directed mutagenesis further revealed that a mutation of the KRK cluster to three alanine residues completely blocked the localization of FEN-1 into the nucleus, whereas mutagenesis of the KKK cluster led to a partial defect of nuclear localization in HeLa cells without observable phenotype in yeast. Therefore, the KRKXXXXXXXXKKK motif may be a bipartite NLS driving the protein into nuclei. Yeast RAD27Delta cells transformed with human mutant M(krk) survived poorly upon methyl methanesulfonate treatment or when they were incubated at an elevated temperature. PMID- 11053419 TI - An ephrin-A-dependent signaling pathway controls integrin function and is linked to the tyrosine phosphorylation of a 120-kDa protein. AB - The Eph family of receptor tyrosine kinases and their ligands, the ephrins, have been implicated in the development of the retinotectal projection. Here, glycosylphosphatidylinositol-anchored A-ephrins are not only expressed in the tectum but also on retinal axons, raising the possibility that they function in this context as receptors. We now show that activation of ephrin-A2 or ephrin-A5 by one of their receptors, ephA3, results in a beta 1-integrin-dependent increased adhesion of ephrin-A-expressing cells to laminin. In the search for an ephrin-A-dependent signaling pathway controlling integrin activation, we identified a 120-kDa raft membrane protein that is tyrosine-phosphorylated specifically after ephrin-A activation. Tyrosine phosphorylation of this protein is not seen after stimulating ephrin-A2-expressing cells with basic fibroblast growth factor, epidermal growth factor, insulin growth factor, or fetal calf serum containing a large set of different growth factors. The role of p120 as a mediator of an ephrin-A-integrin coupling is supported by the finding that inhibiting tyrosine phosphorylation of p120 correlates with an abolishment of the beta 1-dependent cell adhesion. PMID- 11053420 TI - Dimers of beta 2-glycoprotein I mimic the in vitro effects of beta 2-glycoprotein I-anti-beta 2-glycoprotein I antibody complexes. AB - Anti-beta(2)-glycoprotein I antibodies are thought to cause lupus anticoagulant activity by forming bivalent complexes with beta(2)-glycoprotein I (beta(2)GPI). To test this hypothesis, chimeric fusion proteins were constructed of the dimerization domain (apple 4) of factor XI and beta(2)GPI. Both a covalent (apple 4-beta(2)GPI) and a noncovalent (apple 4-C321S-beta(2)GPI) chimer were constructed. As controls, apple 2-beta(2)GPI and apple 4-C321S-beta(2)GPI-W316S, in which beta(2)GPI-W316S is not able to bind to phospholipids, were made. In a phospholipid binding assay, apple 4-beta(2)GPI and apple 4-C321S-beta(2)GPI were able to bind to phospholipids with an affinity 35 times higher than that of plasma-derived beta(2)GPI and apple 2-beta(2)GPI. Apple 4-C321S-beta(2)GPI-W316S did not bind at all. Only apple 4-beta(2)GPI and apple 4-C321S-beta(2)GPI were able to bind to adhered platelets as shown by immunofluorescence. Using the prothrombin time, which was the most responsive coagulation assay, the clotting time was approximately doubled when 200 microg/ml apple 4-beta(2)GPI or apple 4 C321S-beta(2)GPI was added. Addition of 200 microg/ml plasma-derived beta(2)GPI, apple 2-beta(2)GPI, or apple 4-C321S-beta(2)GPI-W316S did not affect clotting time. Clotting time could be corrected with the addition of extra phospholipids, which is indicative for lupus anticoagulant activity. An additional increase in clotting times for apple 4-beta(2)GPI or apple 4-C321S-beta(2)GPI was achieved by the addition of monoclonal antibodies against beta(2)GPI. In conclusion, dimerization of beta(2)GPI explains the in vitro observed effects of beta(2)GPI anti-beta(2)GPI antibody complexes. PMID- 11053421 TI - Syncoilin, a novel member of the intermediate filament superfamily that interacts with alpha-dystrobrevin in skeletal muscle. AB - Dystrophin coordinates the assembly of a complex of structural and signaling proteins that are required for normal muscle function. A key component of the dystrophin protein complex is alpha-dystrobrevin, a dystrophin-associated protein whose absence results in neuromuscular junction defects and muscular dystrophy. To gain further insights into the role of alpha-dystrobrevin in skeletal muscle, we used the yeast two-hybrid system to identify a novel alpha-dystrobrevin binding partner called syncoilin. Syncoilin is a new member of the intermediate filament superfamily and is highly expressed in skeletal and cardiac muscle. In normal skeletal muscle, syncoilin is concentrated at the neuromuscular junction, where it colocalizes and coimmunoprecipitates with alpha-dystrobrevin-1. Expression studies in mammalian cells demonstrate that, while alpha-dystrobrevin and syncoilin associate directly, overexpression of syncoilin does not result in the self-assembly of intermediate filaments. Finally, unlike many components of the dystrophin protein complex, we show that syncoilin expression is up-regulated in dystrophin-deficient muscle. These data suggest that alpha-dystrobrevin provides a link between the dystrophin protein complex and the intermediate filament network at the neuromuscular junction, which may be important for the maintenance and maturation of the synapse. PMID- 11053422 TI - The effector domain of myristoylated alanine-rich C kinase substrate binds strongly to phosphatidylinositol 4,5-bisphosphate. AB - Both the myristoylated alanine-rich protein kinase C substrate protein (MARCKS) and a peptide corresponding to its basic effector domain, MARCKS-(151-175), inhibit phosphoinositide-specific phospholipase C (PLC)-catalyzed hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP(2)) in vesicles (Glaser, M., Wanaski, S., Buser, C. A., Boguslavsky, V., Rashidzada, W., Morris, A., Rebecchi, M., Scarlata, S. F., Runnels, L. W., Prestwich, G. D., Chen, J., Aderem, A., Ahn, J., and McLaughlin, S. (1996) J. Biol. Chem. 271, 26187-26193). We report here that adding 10-100 nm MARCKS-(151-175) to a subphase containing either PLC-delta or beta inhibits hydrolysis of PIP(2) in a monolayer and that this inhibition is due to the strong binding of the peptide to PIP(2). Two direct binding measurements, based on centrifugation and fluorescence, show that approximately 10 nm PIP(2), in the form of vesicles containing 0.01%, 0.1%, or 1% PIP(2), binds 50% of MARCKS (151-175). Both electrophoretic mobility measurements and competition experiments suggest that MARCKS-(151-175) forms an electroneutral complex with approximately 4 PIP(2). MARCKS-(151-175) binds equally well to PI(4,5)P(2) and PI(3,4)P(2). Local electrostatic interactions of PIP(2) with MARCKS-(151-175) contribute to the binding energy because increasing the salt concentration from 100 to 500 mm decreases the binding 100-fold. We hypothesize that the effector domain of MARCKS can bind a significant fraction of the PIP(2) in the plasma membrane, and release the bound PIP(2) upon interaction with Ca(2+)/calmodulin or phosphorylation by protein kinase C. PMID- 11053423 TI - Adrenodoxin reductase-adrenodoxin complex structure suggests electron transfer path in steroid biosynthesis. AB - The steroid hydroxylating system of adrenal cortex mitochondria consists of the membrane-attached NADPH-dependent adrenodoxin reductase (AR), the soluble one electron transport protein adrenodoxin (Adx), and a membrane-integrated cytochrome P450 of the CYP11 family. In the 2.3-A resolution crystal structure of the Adx.AR complex, 580 A(2) of partly polar surface are buried. Main interaction sites are centered around Asp(79), Asp(76), Asp(72), and Asp(39) of Adx and around Arg(211), Arg(240), Arg(244), and Lys(27) of AR, respectively. In particular, the region around Asp(39) defines a new protein interaction site for Adx, similar to those found in plant and bacterial ferredoxins. Additional contacts involve the electron transfer region between the redox centers of AR and Adx and C-terminal residues of Adx. The Adx residues Asp(113) to Arg(115) adopt 3(10)-helical conformation and engage in loose intermolecular contacts within a deep cleft of AR. Complex formation is accompanied by a slight domain rearrangement in AR. The [2Fe-2S] cluster of Adx and the isoalloxazine rings of FAD of AR are 10 A apart suggesting a possible electron transfer route between these redox centers. The AR.Adx complex represents the first structure of a biologically relevant complex between a ferredoxin and its reductase. PMID- 11053424 TI - Cross-talk between G-protein and protein kinase C modulation of N-type calcium channels is dependent on the G-protein beta subunit isoform. AB - The modulation of N-type calcium current by protein kinases and G-proteins is a factor in the fine tuning of neurotransmitter release. We have previously shown that phosphorylation of threonine 422 in the alpha(1B) calcium channel domain I II linker region resulted in a dramatic reduction in somatostatin receptor mediated G-protein inhibition of the channels and that the I-II linker consequently serves as an integration center for cross-talk between protein kinase C (PKC) and G-proteins (Hamid, J., Nelson, D., Spaetgens, R., Dubel, S. J., Snutch, T. P., and Zamponi, G. W. (1999) J. Biol. Chem. 274, 6195-6202). Here we show that opioid receptor-mediated inhibition of N-type channels is affected to a lesser extent compared with that seen with somatostatin receptors, hinting at the possibility that PKC/G-protein cross-talk might be dependent on the G protein subtype. To address this issue, we have examined the effects of four different types of G-protein beta subunits on both wild type and mutant alpha(1B) calcium channels in which residue 422 has been replaced by glutamate to mimic PKC dependent phosphorylation and on channels that have been directly phosphorylated by protein kinase C. Our data show that phosphorylation or mutation of residue 422 antagonizes the effect of Gbeta(1) on channel activity, whereas Gbeta(2), Gbeta(3), and Gbeta(4) are not affected. Our data therefore suggest that the observed cross-talk between G-proteins and protein kinase C modulation of N-type channels is a selective feature of the Gbeta(1) subunit. PMID- 11053425 TI - CARD9 is a novel caspase recruitment domain-containing protein that interacts with BCL10/CLAP and activates NF-kappa B. AB - BCL10/CLAP is an activator of apoptosis and NF-kappaB signaling pathways and has been implicated in B cell lymphomas of mucosa-associated lymphoid tissue. Although its role in apoptosis remains to be determined, BCL10 likely activates NF-kappaB through the IKK complex in response to upstream stimuli. The N-terminal caspase recruitment domain (CARD) of BCL10 has been proposed to function as an activation domain that mediates homophilic interactions with an upstream CARD containing NF-kappaB activator. To identify upstream signaling partners of BCL10, we performed a mammalian two-hybrid analysis and identified CARD9 as a novel CARD containing protein that interacts selectively with the CARD activation domain of BCL10. When expressed in cells, CARD9 binds to BCL10 and activates NF-kappaB. Furthermore, endogenous CARD9 is found associated with BCL10 suggesting that both proteins form a pre-existing signaling complex within cells. CARD9 also self associates and contains extensive coiled-coil motifs that may function as oligomerization domains. We propose here that CARD9 is an upstream activator of BCL10 and NF-kappaB signaling. PMID- 11053426 TI - DNA binding specificity of different STAT proteins. Comparison of in vitro specificity with natural target sites. AB - STAT transcription factors are expressed in many cell types and bind to similar sequences. However, different STAT gene knock-outs show very distinct phenotypes. To determine whether differences between the binding specificities of STAT proteins account for these effects, we compared the sequences bound by STAT1, STAT5A, STAT5B, and STAT6. One sequence set was selected from random oligonucleotides by recombinant STAT1, STAT5A, or STAT6. For another set including many weak binding sites, we quantified the relative affinities to STAT1, STAT5A, STAT5B, and STAT6. We compared the results to the binding sites in natural STAT target genes identified by others. The experiments confirmed the similar specificity of different STAT proteins. Detailed analysis indicated that STAT5A specificity is more similar to that of STAT6 than that of STAT1, as expected from the evolutionary relationships. The preference of STAT6 for sites in which the half-palindromes (TTC) are separated by four nucleotides (N(4)) was confirmed, but analysis of weak binding sites showed that STAT6 binds fairly well to N(3) sites. As previously reported, STAT1 and STAT5 prefer N(3) sites; however, STAT5A, but not STAT1, weakly binds N(4) sites. None of the STATs bound to half-palindromes. There were no specificity differences between STAT5A and STAT5B. PMID- 11053427 TI - Insect immunity. Constitutive expression of a cysteine-rich antifungal and a linear antibacterial peptide in a termite insect. AB - Two novel antimicrobial peptides, which we propose to name termicin and spinigerin, have been isolated from the fungus-growing termite Pseudacanthotermes spiniger (heterometabole insect, Isoptera). Termicin is a 36-amino acid residue antifungal peptide, with six cysteines arranged in a disulfide array similar to that of insect defensins. In contrast to most insect defensins, termicin is C terminally amidated. Spinigerin consists of 25 amino acids and is devoid of cysteines. It is active against bacteria and fungi. Termicin and spinigerin show no obvious sequence similarities with other peptides. Termicin is constitutively present in hemocyte granules and in salivary glands. The presence of termicin and spinigerin in unchallenged termites contrasts with observations in evolutionary recent insects or insects undergoing complete metamorphosis, in which antimicrobial peptides are induced in the fat body and released into the hemolymph after septic injury. PMID- 11053428 TI - The kinase activation loop is the key to mixed lineage kinase-3 activation via both autophosphorylation and hematopoietic progenitor kinase 1 phosphorylation. AB - We have demonstrated previously that Cdc42 induced MLK-3 homodimerization leads to both autophosphorylation and activation of MLK-3 and postulated that autophosphorylation is an intermediate step of MLK-3 activation following its dimerization. In this report we sought to refine further the mechanism of MLK-3 activation and study the role of the putative kinase activation loop in MLK-3 activation. First we mutated the three potential phosphorylation sites in MLK-3 putative activation loop to alanine in an effort to abrogate MLK-3 autophosphorylation. Mutant T277A displayed almost no autophosphorylation activity and was nearly nonfunctional; mutant S281A, that displayed a low level of autophosphorylation, only slightly activated its downstream targets, whereas the T278A mutant, that exhibited autophosphorylation comparable to that of the wild type, was almost fully functional. Thus, these residues within the activation loop are critical for MLK-3 autophosphorylation and activation. In addition, when the Thr277 and Ser281 residues were mutated to negatively charged glutamic acid to mimic phosphorylated serine/threonine residues, the resulting mutants were fully functional, implying that these two residues may serve as the autophosphorylation sites. Interestingly, HPK1 also phosphorylated MLK-3 activation loop in vitro, and Ser281 was found to be the major phosphorylation site, indicating that HPK1 also activates MLK-3 via phosphorylation of the kinase activation loop. PMID- 11053429 TI - Orf135 from Escherichia coli Is a Nudix hydrolase specific for CTP, dCTP, and 5 methyl-dCTP. AB - Orf135 from Escherichia coli is a new member of the Nudix (nucleoside diphosphate linked to some other moiety, x) hydrolase family of enzymes with substrate specificity for CTP, dCTP, and 5-methyl-dCTP. The gene has been cloned for overexpression, and the protein has been overproduced, purified, and characterized. Orf135 is most active on 5-methyl-dCTP (k(cat)/K(m) = 301,000 M( 1) s(-1)), followed by CTP (k(cat)/K(m) = 47,000 M(-1) s(-1)) and dCTP (k(cat)/K(m) = 18,000 M(-1) s(-1)). Unlike other nucleoside triphosphate pyrophophohydrolases of the Nudix hydrolase family discovered thus far, Orf135 is highly specific for pyrimidine (deoxy)nucleoside triphosphates. Like other Nudix hydrolases, the enzyme cleaves its substrates to produce a nucleoside monophosphate and inorganic pyrophosphate, has an alkaline pH optimum, and requires a divalent metal cation for catalysis, with magnesium yielding optimal activity. Because of the nature of its substrate specificity, Orf135 may play a role in pyrimidine biosynthesis, lipid biosynthesis, and in controlling levels of 5-methyl-dCTP in the cell. PMID- 11053430 TI - Molecular properties of Zic proteins as transcriptional regulators and their relationship to GLI proteins. AB - Zic family genes encode zinc finger proteins, which play important roles in vertebrate development. The zinc finger domains are highly conserved between Zic proteins and show a notable homology to those of Gli family proteins. In this study, we investigated the functional properties of Zic proteins and their relationship to the GLI proteins. We first established an optimal binding sequence for Zic1, Zic2, and Zic3 proteins by electrophoretic mobility shift assay-based target selection and mutational analysis. The selected sequence was almost identical to the GLI binding sequence. However, the binding affinity was lower than that of GLI. Consistent results were obtained in reporter assays, in which transcriptional activation by Zic proteins was less dependent on the GLI binding sequence than GLI1. Moreover, Zic proteins activated a wide range of promoters irrespective of the presence of a GLI binding sequence. When Zic and GLI proteins were cotransfected into cultured cells, Zic proteins enhanced or suppressed sequence-dependent, GLI-mediated transactivation depending on cell type. Taken together, these results suggest that Zic proteins may act as transcriptional coactivators and that their function may be modulated by the GLI proteins and possibly by other cell type-specific cofactors. PMID- 11053431 TI - Crystal structure of paprika ferredoxin-NADP+ reductase. Implications for the electron transfer pathway. AB - cDNA of Capsicum annuum Yolo Wonder (paprika) has been prepared from total cellular RNA, and the complete gene encoding paprika ferredoxin-NADP(+) reductase (pFNR) precursor was sequenced and cloned from this cDNA. Fusion to a T7 promoter allowed expression in Escherichia coli. Both native and recombinant pFNR were purified to homogeneity and crystallized. The crystal structure of pFNR has been solved by Patterson search techniques using the structure of spinach ferredoxin NADP(+) reductase as search model. The structure was refined at 2.5-A resolution to a crystallographic R-factor of 19.8% (R(free) = 26.5%). The overall structure of pFNR is similar to other members of the ferredoxin-NADP(+) reductase family, the major differences concern a long loop (residues 167-177) that forms part of the FAD binding site and some of the variable loops in surface regions. The different orientation of the FAD binding loop leads to a tighter interaction between pFNR and the adenine moiety of FAD. The physiological redox partners [2Fe 2S]-ferredoxin I and NADP(+) were modeled into the native structure of pFNR. The complexes reveal a protein-protein interaction site that is consistent with existing biochemical data and imply possible orientations for the side chain of tyrosine 362, which has to be displaced by the nicotinamide moiety of NADP(+) upon binding. A reasonable electron transfer pathway could be deduced from the modeled structures of the complexes. PMID- 11053432 TI - The delta subunit of type 6 phosphodiesterase reduces light-induced cGMP hydrolysis in rod outer segments. AB - The delta subunit of the rod photoreceptor PDE has previously been shown to copurify with the soluble form of the enzyme and to solubilize the membrane-bound form (). To determine the physiological effect of the delta subunit on the light response of bovine rod outer segments, we measured the real time accumulation of the products of cGMP hydrolysis in a preparation of permeablized rod outer segments. The addition of delta subunit GST fusion protein (delta-GST) to this preparation caused a reduction in the maximal rate of cGMP hydrolysis in response to light. The maximal reduction of the light response was about 80%, and the half maximal effect occurred at 385 nm delta subunit. Several experiments suggest that this effect was not due to the effects of delta-GST on transducin or rhodopsin kinase. Immunoblots demonstrated that exogenous delta-GST solubilized the majority of the PDE in ROS but did not affect the solubility of transducin. Therefore, changes in the solubility of transducin cannot account for the effects of delta-GST in the pH assay. The reduction in cGMP hydrolysis was independent of ATP, which indicates that it was not due to effects of delta-GST on rhodopsin kinase. In addition to the effect on cGMP hydrolysis, the delta-GST fusion protein slowed the turn-off of the system. This is probably due, at least in part, to an observed reduction in the GTPase rate of transducin in the presence of delta-GST. These results demonstrate that delta-GST can modify the activity of the phototransduction cascade in preparations of broken rod outer segments, probably due to a functional uncoupling of the transducin to PDE step of the signal transduction cascade and suggest that the delta subunit may play a similar role in the intact outer segment. PMID- 11053433 TI - Transcriptional regulation of apolipoprotein C-III gene expression by the orphan nuclear receptor RORalpha. AB - Triglyceride-rich remnant lipoproteins are considered as major risk factors contributing to the pathogenesis of atherosclerosis. Because apolipoprotein (apo) C-III is a major determinant of plasma triglyceride and remnant lipoprotein metabolism, it is important to understand how the expression of this gene is regulated. In the present study, we identified the orphan nuclear receptor RORalpha1 as a regulator of human and mouse apo C-III gene expression. Plasma triglyceride and apo C-III protein concentrations in staggerer (sg/sg) mice, homozygous for a deletion in the RORalpha gene, were significantly lower than in wild type littermates. The lowered plasma apo C-III levels were associated with reduced apo C-III mRNA levels in liver and intestine of sg/sg mice. Transient transfection experiments in human hepatoma HepG2, human colonic CaCO2, and rabbit kidney RK13 cells demonstrated that overexpression of the human RORalpha1 isoform specifically increases human apo C-III promoter activity, indicating that RORalpha1 enhances human apo C-III gene transcription. RORalpha1 response elements were mapped by promoter deletion analysis and gel shift experiments to two AGGTCA half-sites located at positions -83/-78 (within the C3P site) and -23/ 18 (downstream of the TATA box) in the human apo C-III promoter, with the -23/-18 site exhibiting the highest binding affinity. Transfection of site-directed mutated constructs in HepG2 cells indicated that the RORalpha1 effect is predominantly mediated by the -23/-18 site. This site is conserved in the mouse apo C-III gene promoter. Moreover, RORalpha binds to the equivalent mouse site and activates constructs containing three copies of the mouse site cloned in front of an heterologous promoter. Taken together, our data identify RORalpha as a transcriptional regulator of apo C-III gene expression, providing a novel, physiological role for RORalpha1 in the regulation of genes controlling triglyceride metabolism. PMID- 11053434 TI - The steroidogenic acute regulatory protein homolog MLN64, a late endosomal cholesterol-binding protein. AB - MLN64 is a transmembrane protein that shares homology with the cholesterol binding domain (START domain) of the steroidogenic acute regulatory protein. The steroidogenic acute regulatory protein is located in the inner membrane of mitochondria, where it facilitates cholesterol import into the mitochondria. Crystallographic analysis showed that the START domain of MLN64 is a cholesterol binding domain. The present work was undertaken to determine which step of the intracellular cholesterol pathway MLN64 participates in. Using immunocytofluorescence, MLN64 colocalizes with LBPA, a lipid found specifically in late endosomes. Electron microscopy indicates that MLN64 is restricted to the limiting membrane of late endosomes. Microinjection or endocytosis of specific antibodies shows that the START domain of MLN64 is cytoplasmic. Deletion and mutagenesis experiments demonstrate that the amino-terminal part of MLN64 is responsible for its addressing. Although this domain does not contain conventional dileucine- or tyrosine-based targeting signals, we show that a dileucine motif (Leu(66)-Leu(67)) and a tyrosine residue (Tyr(89)) are critical for the targeting or the proper folding of the molecule. Finally, MLN64 colocalizes with cholesterol and Niemann Pick C1 protein in late endosomes. However, complementation assays show that MLN64 is not involved in the Niemann Pick C2 disease which, results in cholesterol lysosomal accumulation. Together, our results show that MLN64 plays a role at the surface of the late endosomes, where it might shuttle cholesterol from the limiting membrane to cytoplasmic acceptor(s). PMID- 11053435 TI - Role of mitochondria and caspases in vitamin D-mediated apoptosis of MCF-7 breast cancer cells. AB - Vitamin D(3) compounds are currently in clinical trials for human breast cancer and offer an alternative approach to anti-hormonal therapies for this disease. 1alpha,25-Dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)), the active form of vitamin D(3), induces apoptosis in breast cancer cells and tumors, but the underlying mechanisms are poorly characterized. In these studies, we focused on the role of caspase activation and mitochondrial disruption in 1alpha,25(OH)(2)D(3)-mediated apoptosis in breast cancer cells (MCF-7) in vitro. The effect of 1alpha,25(OH)(2)D(3) on MCF-7 cells was compared with that of tumor necrosis factor alpha, which induces apoptosis via a caspase-dependent pathway. Our major findings are that 1alpha,25(OH)(2)D(3) induces apoptosis in MCF-7 cells by disruption of mitochondrial function, which is associated with Bax translocation to mitochondria, cytochrome c release, and production of reactive oxygen species. Moreover, we show that Bax translocation and mitochondrial disruption do not occur after 1alpha,25(OH)(2)D(3) treatment of a MCF-7 cell clone selected for resistance to 1alpha,25(OH)(2)D(3)-mediated apoptosis. These mitochondrial effects of 1alpha,25(OH)(2)D(3) do not require caspase activation, since they are not blocked by the cell-permeable caspase inhibitor z-Val-Ala-Asp fluoromethylketone. Although caspase inhibition blocks 1alpha,25(OH)(2)D(3) mediated events downstream of mitochondria such as poly(ADP-ribose) polymerase cleavage, external display of phosphatidylserine, and DNA fragmentation, MCF-7 cells still execute apoptosis in the presence of z-Val-Ala-Asp fluoromethylketone, indicating that the commitment to 1alpha,25(OH)(2)D(3) mediated cell death is caspase-independent. PMID- 11053436 TI - Inhibition of angiogenesis by a mouse sprouty protein. AB - Sprouty negatively modulates branching morphogenesis in the Drosophila tracheal system. To address the role of mammalian Sprouty homologues in angiogenesis, another form of branching morphogenesis, a recombinant adenovirus engineered to express murine Sprouty-4 selectively in endothelial cells, was injected into the sinus venosus of embryonic day 9.0 cultured mouse embryos. Sprouty-4 expression inhibited branching and sprouting of small vessels, resulting in abnormal embryonic development. In vitro, Sprouty-4 inhibited fibroblast growth factor and vascular endothelial cell growth factor-mediated cell proliferation and migration and prevented basic fibroblast growth factor and vascular endothelial cell growth factor-induced MAPK phosphorylation in endothelial cells, indicating inhibition of tyrosine kinase-mediated signaling pathways. The ability of constitutively activated mutant Ras(L61) to rescue Sprouty-4 inhibition of MAPK phosphorylation suggests that Sprouty inhibits receptor tyrosine kinase signaling upstream of Ras. Thus, Sprouty may regulate angiogenesis in normal and disease processes by modulating signaling by endothelial tyrosine kinases. PMID- 11053437 TI - Evidence for direct interaction between Sprouty and Cbl. AB - Sprouty (SPRY) was first identified in a genetic screen in Drosophila as an antagonist of fibroblast and epidermal growth factor receptors and Sevenless signaling, seemingly by inhibiting the receptor tyrosine kinase (RTK)/Ras/MAPK pathway. To date, four mammalian Sprouty genes have been identified; the primary sequences of the gene products share a well conserved cysteine-rich C-terminal domain with their Drosophila counterpart. The N-terminal regions do not, however, exhibit a large degree of homology. This study was aimed at identifying proteins with which human SPRY2 (hSPRY2) interacts in an attempt to understand the mechanism by which Sprouty proteins exert their down-regulatory effects. Here, we demonstrate that hSPRY2 associates directly with c-Cbl, a known down-regulator of RTK signaling. A short sequence in the N terminus of hSPRY2 was found to bind directly to the Ring finger domain of c-Cbl. Parallel binding was apparent between the Drosophila homologs of Sprouty and Cbl, with cross-species associations occurring at least in vitro. Coexpression of hSPRY2 abrogated an increase in the rate of epidermal growth factor receptor internalization induced by c-Cbl, whereas a mutant hSPRY2 protein unable to bind c-Cbl showed no such effect. Our results suggest that one function of hSPRY2 in signaling processes downstream of RTKs may be to modulate c-Cbl physiological function such as that seen with receptor-mediated endocytosis. PMID- 11053438 TI - Intracellular calcium mobilization induces immediate early gene pip92 via Src and mitogen-activated protein kinase in immortalized hippocampal cells. AB - Regulation of intracellular calcium levels plays a central role in cell survival, proliferation, and differentiation. A cell-permeable, tumor-promoting thapsigargin elevates the intracellular calcium levels by inhibiting endoplasmic reticulum Ca(2+)-ATPase. The Src-tyrosine kinase family is involved in a broad range of cellular responses ranging from cell growth and cytoskeletal rearrangement to differentiation. The immediate early gene pip92 is induced in neuronal cell death as well as cell growth and differentiation. To resolve the molecular mechanism of cell growth by intracellular calcium mobilization, we have examined the effect of thapsigargin and subsequent intracellular calcium influx on pip92 expression in immortalized rat hippocampal H19-7 cells. An increase of intracellular calcium ion levels induced by thapsigargin stimulated the expression of pip92 in H19-7 cells. Transient transfection of the cells with kinase-inactive mitogen-activated protein kinase kinase (MEK) and Src kinase or pretreatment with the chemical MEK inhibitor PD98059 significantly inhibited pip92 expression induced by thapsigargin. When constitutively active v-Src or MEK was overexpressed, the transcriptional activity of the pip92 gene was markedly increased. Dominant inhibitory Raf-1 blocked the transcriptional activity of pip92 induced by thapsigargin. The transcription factor Elk1 is activated during thapsigargin-induced pip92 expression. Taken together, these results suggest that an increase of intracellular calcium ion levels by thapsigargin stimulates the pip92 expression via Raf-MEK-extracellular signal-regulated protein kinase- as well as Src kinase-dependent signaling pathways. PMID- 11053439 TI - Crystal structure of the iron-dependent regulator from Mycobacterium tuberculosis at 2.0-A resolution reveals the Src homology domain 3-like fold and metal binding function of the third domain. AB - Iron-dependent regulators are primary transcriptional regulators of virulence factors and iron scavenging systems that are important for infection by several bacterial pathogens. Here we present the 2.0-A crystal structure of the wild type iron-dependent regulator from Mycobacterium tuberculosis in its fully active holorepressor conformation. Clear, unbiased electron density for the Src homology domain 3-like third domain, which is often invisible in structures of iron dependent regulators, was revealed by density modification and averaging. This domain is one of the rare examples of Src homology domain 3-like folds in bacterial proteins, and, in addition, displays a metal binding function by contributing two ligands, one Glu and one Gln, to the pentacoordinated cobalt atom at metal site 1. Both metal sites are fully occupied, and tightly bound water molecules at metal site 1 ("Water 1") and metal site 2 ("Water 2") are identified unambiguously. The main chain carbonyl of Leu4 makes an indirect interaction with the cobalt atom at metal site 2 via Water 2, and the adjacent residue, Val5, forms a rare gamma turn. Residues 1-3 are well ordered and make numerous interactions. These ordered solvent molecules and the conformation and interactions of the N-terminal pentapeptide thus might be important in metal dependent activation. PMID- 11053440 TI - Urokinase-type plasminogen activator receptor (CD87) is a ligand for integrins and mediates cell-cell interaction. AB - Binding of urokinase-type plasminogen activator (uPA) to its receptor (uPAR/CD87) regulates cellular adhesion, migration, and tumor cell invasion. However, it is unclear how glycosyl phosphatidylinositol-anchored uPAR, which lacks a transmembrane structure, mediates signal transduction. It has been proposed that uPAR forms cis-interactions with integrins as an associated protein and thereby transduces proliferative or migratory signals to cells upon binding of uPA. We provide evidence that soluble uPAR (suPAR) specifically binds to integrins alpha4beta1, alpha6beta1, alpha9beta1, and alphavbeta3 on Chinese hamster ovary cells in a cation-dependent manner. Anti-integrin and anti-uPAR antibodies effectively block binding of suPAR to these integrins. Binding of suPAR to alpha4beta1 and alphavbeta3 is blocked by known soluble ligands and by the integrin mutations that inhibit ligand binding. These results suggest that uPAR is an integrin ligand rather than, or in addition to, an integrin-associated protein. In addition, we demonstrate that glycosyl phosphatidylinositol-anchored uPAR on the cell surface specifically binds to integrins on the apposing cells, suggesting that uPAR-integrin interaction may mediate cell-cell interaction (trans-interaction). These previously unrecognized uPAR-integrin interactions may allow uPAR to transduce signals through the engaged integrin without a hypothetical transmembrane adapter and may provide a potential therapeutic target for control of inflammation and cancer. PMID- 11053441 TI - The toxoplasma micronemal protein MIC4 is an adhesin composed of six conserved apple domains. AB - The initial stage of invasion by apicomplexan parasites involves the exocytosis of the micronemes-containing molecules that contribute to host cell attachment and penetration. MIC4 was previously described as a protein secreted by Toxoplasma gondii tachyzoites upon stimulation of micronemes exocytosis. We have microsequenced the mature protein, purified after discharge from micronemes and cloned the corresponding gene. The deduced amino acid sequence of MIC4 predicts a 61-kDa protein that contains 6 conserved apple domains. Apple domains are composed of six spacely conserved cysteine residues which form disulfide bridges and are also present in micronemal proteins from two closely related apicomplexan parasites, Sarcocystis muris and Eimeria species, and several mammalian serum proteins, including kallikrein. Here we show that MIC4 localizes in the micronemes of all the invasive forms of T. gondii, tachyzoites, bradyzoites, sporozoites, and merozoites. The protein is proteolytically processed both at the N and the C terminus only upon release from the organelle. MIC4 binds efficiently to host cells, and the adhesive motif maps in the most C-terminal apple domain. PMID- 11053442 TI - Degradation of nucleosome-associated centromeric histone H3-like protein CENP-A induced by herpes simplex virus type 1 protein ICP0. AB - Cells infected by herpes simplex virus type 1 in the G2 phase of the cell cycle become stalled at an unusual stage of mitosis defined as pseudoprometaphase. This block correlates with the viral immediate-early protein ICP0-induced degradation of the centromere protein CENP-C. However, the observed pseudoprometaphase phenotype of infected mitotic cells suggests that the stability of other centromere proteins may also be affected. Here, we demonstrate that ICP0 also induces the proteasome-dependent degradation of the centromere protein CENP-A. By a series of Western blot and immunofluorescence experiments we show that the endogenous 17-kDa CENP-A and an exogenous tagged version of CENP-A are lost from centromeres and degraded in infected and transfected cells as a result of ICP0 expression. CENP-A is a histone H3-like protein associated with nucleosome structures in the inner plate of the kinetochore. Unlike fully transcribed lytic viral DNA, the transcriptionally repressed latent herpes simplex virus type 1 genome has been reported to have a nucleosomal structure similar to that of cellular chromatin. Because ICP0 plays an essential part in controlling the balance between the lytic and latent outcomes of infection, the ICP0-induced degradation of CENP-A is an intriguing feature connecting different aspects of viral and/or cellular genome regulation. PMID- 11053443 TI - A role for the polyproline domain of p53 in its regulation by Mdm2. AB - The p53 protein plays a key role in the cellular response to stress by inducing cell growth arrest or apoptosis. The polyproline region of p53 has been shown to be important for its growth suppression activity. p53 protein lacking the polyproline region has impaired apoptotic activity and altered specificity for certain apoptotic target genes. Here we describe the role of this region in the regulation of p53 by its inhibitor Mdm2. p53 lacking the polyproline region was identified to be more susceptible to inhibition by Mdm2. Furthermore, the absence of this region renders p53 more accessible to ubiquitination, nuclear export, and Mdm2-mediated degradation. This increased sensitivity to Mdm2 results from an enhanced affinity of Mdm2 toward p53 lacking the polyproline region. Our results provide a new explanation for the impaired growth suppression activity of p53 lacking this region. The polyproline region is proposed to be important in the modulation of the inhibitory effects of Mdm2 on p53 activities and stability. PMID- 11053444 TI - Crystal structure of the ligand-binding domain of the ultraspiracle protein USP, the ortholog of retinoid X receptors in insects. AB - The major postembryonic developmental events happening in insect life, including molting and metamorphosis, are regulated and coordinated temporally by pulses of ecdysone. The biological activity of this steroid hormone is mediated by two nuclear receptors: the ecdysone receptor (EcR) and the Ultraspiracle protein (USP). The crystal structure of the ligand-binding domain from the lepidopteran Heliothis virescens USP reported here shows that the loop connecting helices H1 and H3 precludes the canonical agonist conformation. The key residues that stabilize this unique loop conformation are strictly conserved within the lepidopteran USP family. The presence of an unexpected bound ligand that drives an unusual antagonist conformation confirms the induced-fit mechanism accompanying the ligand binding. The ligand-binding pocket exhibits a retinoid X receptor-like anchoring part near a conserved arginine, which could interact with a USP ligand functional group. The structure of this receptor provides the template for designing inhibitors, which could be utilized as a novel type of environmentally safe insecticides. PMID- 11053445 TI - Phosphorylation of Wzc, a tyrosine autokinase, is essential for assembly of group 1 capsular polysaccharides in Escherichia coli. AB - Wzc proteins are tyrosine autokinases. They are found in some important bacterial pathogens of humans and livestock as well as plant-associated bacteria, and are often encoded within gene clusters determining synthesis and assembly of capsular and extracellular polysaccharides. Autophosphorylation of Wzc(cps) is essential for assembly of the serotype K30 group 1 capsule in Escherichia coli O9a:K30, although a genetically unlinked Wzc(cps)-homologue (Etk) can also participate with low efficiency. While autophosphorylation of Wzc(cps) is required for assembly of high molecular weight K30 capsular polysaccharide, it is not essential for either the synthesis of the K30 repeat units or for activity of the K30 polymerase enzyme. Paradoxically, the cognate phosphotyrosine protein phosphatase for Wzc(cps), Wzb(cps), is also required for capsule expression. The tyrosine-rich domain at the C terminus of Wzc(cps) was identified as the site of phosphorylation and autophosphorylation of Wzc requires a functional Walker A motif. Intermolecular transphosphorylation of Wzc(cps) was detected in strains expressing a combination of mutant Wzc(cps) derivatives. The N- and C-terminal domains of Wzc(cps) were expressed independently to mimic the situation found naturally in Gram-positive bacteria. In this format, both domains were required for phosphorylation of the Wzc(cps) C terminus, and for capsule assembly. Regulation by a post-translational phosphorylation event represents a new dimension in the assembly of bacterial cell-surface polysaccharides. PMID- 11053446 TI - Sp1 phosphorylation regulates apoptosis via extracellular FasL-Fas engagement. AB - Apoptosis of smooth muscle cells (SMC) in atherosclerotic vessels can destabilize the atheromatus plaque and result in rupture, thrombosis, and sudden death. In efforts to understand the molecular processes regulating apoptosis in this cell type, we have defined a novel mechanism involving the ubiquitously expressed transcription factor Sp1. Subtypes of SMC expressing abundant levels of Sp1 produce the death agonist, Fas ligand (FasL) and undergo greater spontaneous apoptosis. Sp1 activates the FasL promoter via a distinct nucleotide recognition element whose integrity is crucial for inducible expression. Inducible FasL promoter activation is also inhibited by a dominant-negative form of Sp1. Increased SMC apoptosis is preceded by Sp1 phosphorylation, increased FasL transcription, and the autocrine/paracrine engagement of FasL with its cell surface receptor, Fas. Inducible FasL transcription and apoptosis are blocked by dominant-negative protein kinase C-zeta, whose wild-type counterpart phosphorylates Sp1. Thus, Sp1 phosphorylation is a proapoptotic transcriptional event in vascular SMC and, given the wide distribution of this housekeeping transcription factor, may be a common regulatory theme in apoptotic signal transduction. PMID- 11053447 TI - The Fe/S assembly protein IscU behaves as a substrate for the molecular chaperone Hsc66 from Escherichia coli. AB - IscU, a NifU-like Fe/S-escort protein, binds to and stimulates the ATPase activity of Hsc66, a hsp70-type molecular chaperone. We present evidence that stimulation arises from interactions of IscU with the substrate-binding site of Hsc66. IscU inhibited the ability of Hsc66 to suppress the aggregation of the denatured model substrate proteins rhodanese and citrate synthase, and calorimetric and surface plasmon resonance measurements showed that ATP destabilizes Hsc66.IscU complexes in a manner expected for hsp70-substrate complexes. Studies on the interaction of IscU with Hsc66 truncation mutants further showed that IscU does not bind the isolated ATPase domain of Hsc66 but does bind and stimulate a mutant containing the ATPase domain and substrate binding beta-sandwich subdomain. These results support a role for IscU as a substrate for Hsc66 and suggest a specialized function for Hsc66 in the assembly, stabilization, or transfer of Fe/S clusters formed on IscU. PMID- 11053448 TI - Identification of SWI.SNF complex subunit BAF60a as a determinant of the transactivation potential of Fos/Jun dimers. AB - Fos family proteins form stable heterodimers with Jun family proteins, and each heterodimer shows distinctive transactivating potential for regulating cellular growth, differentiation, and development via AP-1 binding sites. However, the molecular mechanism underlying dimer specificity and the molecules that facilitate transactivation remain undefined. Here, we show that BAF60a, a subunit of the SWI.SNF chromatin remodeling complex, is a determinant of the transactivation potential of Fos/Jun dimers. BAF60a binds to a specific subset of Fos/Jun heterodimers using two different interfaces for c-Fos and c-Jun, respectively. Only when the functional SWI.SNF complex is present, can c-Fos/c Jun (high affinity to BAF60a) but not Fra-2/JunD (no affinity to BAF60a) induce the endogenous AP-1-regulated genes such as collagenase and c-met. These results indicate that a specific subset of Fos/Jun dimers recruits SWI.SNF complex via BAF60a to initiate transcription. PMID- 11053449 TI - Triptolide and chemotherapy cooperate in tumor cell apoptosis. A role for the p53 pathway. AB - Triptolide (PG490), a diterpene triepoxide, is a potent immunosuppressive agent extracted from the Chinese herb Tripterygium wilfordii. We have previously shown that triptolide blocks NF-kappaB activation and sensitizes tumor necrosis factor (TNF-alpha)-resistant tumor cell lines to TNF-alpha-induced apoptosis. We show here that triptolide enhances chemotherapy-induced apoptosis. In triptolide treated cells, the expression of p53 increased but the transcriptional function of p53 was inhibited, and we observed a down-regulation of p21(waf1/cip1), a p53 responsive gene. The increase in levels of the p53 protein was mediated by enhanced translation of the p53 protein. Additionally, triptolide induced accumulation of cells in S phase and blocked doxorubicin-mediated accumulation of cells in G(2)/M and doxorubicin-mediated induction of p21. Our data suggest that triptolide, by blocking p21-mediated growth arrest, enhances apoptosis in tumor cells. PMID- 11053450 TI - Characterization of tumor-associated Chk2 mutations. AB - The integrity of the DNA damage response pathway is essential for prevention of neoplastic transformation. Several proteins involved in this pathway including p53, BRCA1, and ATM are frequently mutated in human cancer. Checkpoint kinase 2 (Chk2) is a DNA damage-activated protein kinase that lies downstream of ATM in this pathway. Recently, heterozygous germline mutations in Chk2 have been identified in a subset of patients with Li-Fraumeni syndrome, a highly penetrant familial cancer phenotype, suggesting that Chk2 is a tumor suppressor gene. In this study, we have reported the biochemical characterization of the four tumor associated Chk2 mutants. Two of the reported Chk2 mutations identified in Li Fraumeni syndrome result in loss of Chk2 kinase activity. Whereas one mutation within the Chk2 forkhead homology-associated (FHA) domain, R145W, retains some basal kinase activity, this mutant cannot be phosphorylated at an ATM-dependent phosphorylation site (Thr-68) and cannot be activated following gamma radiation. Wild-type Chk2 exists mainly in a protein complex of M(r) approximately 200,000 whereas the R145W mutant forms a larger, presumably inactive complex in the cell. The other FHA domain mutant, I157T, behaves as wild-type Chk2 in all the assays used here. Because the FHA domain is involved in protein-protein interactions, this mutation may affect associations of Chk2 with other proteins. Additionally, we have shown that Chk2 can also be inactivated by down-regulation of its expression in cancer cells. Thus, Chk2 may be inactivated by multiple mechanisms in the cell. PMID- 11053451 TI - DNA gyrase-mediated wrapping of the DNA strand is required for the replication fork arrest by the DNA gyrase-quinolone-DNA ternary complex. AB - The ability of DNA gyrase (Gyr) to wrap the DNA strand around itself allows Gyr to introduce negative supercoils into DNA molecules. It has been demonstrated that the deletion of the C-terminal DNA-binding domain of the GyrA subunit abolishes the ability of Gyr to wrap the DNA strand and catalyze the supercoiling reaction (Kampranis, S. C., and Maxwell, A. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 14416-14421). By using this mutant Gyr, Gyr (A59), we have studied effects of Gyr-mediated wrapping of the DNA strand on its replicative function and its interaction with the quinolone antibacterial drugs. We find that Gyr (A59) can support oriC DNA replication in vitro. However, Gyr (A59)-catalyzed decatenation activity is not efficient enough to complete the decatenation of replicating daughter DNA molecules. As is the case with topoisomerase IV, the active cleavage and reunion activity of Gyr is required for the formation of the ternary complex that can arrest replication fork progression in vitro. Although the quinolone drugs stimulate the covalent Gyr (A59)-DNA complex formation, the Gyr (A59) quinolone-DNA ternary complexes do not arrest the progression of replication forks. Thus, the quinolone-induced covalent topoisomerase-DNA complex formation is necessary but not sufficient to cause the inhibition of DNA replication. We also assess the stability of ternary complexes formed with Gyr (A59), the wild type Gyr, or topoisomerase IV. The ternary complexes formed with Gyr (A59) are more sensitive to salt than those formed with either the wild type Gyr or topoisomerase IV. Furthermore, a competition experiment demonstrates that the ternary complexes formed with Gyr (A59) readily disassociate from the DNA, whereas the ternary complexes formed with either the wild type Gyr or topoisomerase IV remain stably bound. Thus, Gyr-mediated wrapping of the DNA strand is required for the formation of the stable Gyr-quinolone-DNA ternary complex that can arrest replication fork progression. PMID- 11053452 TI - Plant uptake of radiocaesium: a review of mechanisms, regulation and application. AB - Soil contamination with radiocaesium (Cs) has a long-term radiological impact because it is readily transferred through food chains to human beings. Plant uptake is the major pathway for the migration of radiocaesium from soil to human diet. The plant-related factors that control the uptake of radiocaesium are reviewed. Of these, K supply exerts the greatest influence on Cs uptake from solution. It appears that the uptake of radiocaesium is operated mainly by two transport pathways on plant root cell membranes, namely the K(+) transporter and the K(+) channel pathway. Cationic interactions between K and Cs on isolated K channels or K transporters are in agreement with studies using intact plants. The K(+) transporter functioning at low external potassium concentration (often <0.3 mM) shows little discrimination against Cs(+), while the K(+) channel is dominant at high external potassium concentration with high discrimination against Cs(+). Caesium has a high mobility within plants. Although radiocaesium is most likely taken up by the K transport systems within the plant, the Cs:K ratio is not uniform within the plant. Difference in internal Cs concentration (when expressed on a dry mass basis) may vary by a factor of 20 between different plant species grown under similar conditions. Phytoremediation may be a possible option to decontaminate radiocaesium-contaminated soils, but its major limitation is that it takes an excessively long time (tens of years) and produces large volumes of waste. PMID- 11053453 TI - Increased leakiness of the tetracycline-inducible Triple-Op promoter in dividing cells renders it unsuitable for high inducible levels of a dominant negative CDC2aAt gene. AB - A tetracycline-inducible promoter system was used to generate transgenic tobacco plants that confer inducible expression of the wild type or a dominant negative allele of the gene coding for the cyclin-dependent kinase (CDK) of Arabidopsis thaliana CDC2aAt. Although the total extractable CDK activity was doubled, the induced expression of the wild-type CDC2aAt did not correlate with any change of the cell cycle kinetics. An increase of CDK activity upon CDC2aAt expression was only seen in dividing cell populations, demonstrating that CDC2aAt expression itself is not sufficient to induce CDK activation. Induced expression of the dominant negative CDC2aAt.N146 correlated with a reduction of CDK activity to 66% of the level found in non-induced cells. This decrease was not sufficient to block cell division. The isolation of plants showing only low inducible levels of CDC2aAt.N146 suggests that a counterselection against strong inducible lines had occurred. Accordingly, Triple-Op promoter activity was found in dividing cells in the absence of tetracycline. PMID- 11053454 TI - The translational apparatus of Tortula ruralis: polysomal retention of transcripts encoding the ribosomal proteins RPS14, RPS16 and RPL23 in desiccated and rehydrated gametophytes. AB - Tortula ruralis (Syntrichia ruralis) is an important model system for the study of plant vegetative desiccation tolerance. One of the most intriguing aspects of desiccation-tolerant plants is the maintenance of key cellular components in stable and viable forms in the desiccated state, particularly those related to the translational apparatus (i.e. ribosomes and ribosomal RNAs). This study investigated the third integral component of the translational apparatus, the ribosomal proteins. Three T. ruralis cDNAs encoding predicted polypeptides with significant similarity to ribosomal proteins were isolated from a cDNA expression library derived from the polysomal, messenger ribonucleoprotein particle (mRNP) fraction of desiccated gametophytes; Rps14 and Rps16 encode the small-subunit ribosomal proteins RPS14 and RPS16, respectively, and Rpl23 encodes the large subunit ribosomal protein RPL23. RPS14, RPS16 and RPL23, the deduced polypeptides, have predicted molecular masses of 14.4 kDa, 16.2 kDa and 14.9 kDa and predicted pI's of 11.08, 10.34 and 10. 67, respectively. Phylogenetic analysis of the deduced amino acid sequences demonstrated that each of the T. ruralis proteins is most similar to ribosomal proteins from higher plants even though RPS14 and RPL23 show high divergence from their other plant counterparts. RNA blot hybridizations of RNAs present within the polysomal mRNP fraction (i.e. the 100 Kxg pellet) demonstrated that Rps14, Rps16 and Rpl23 are expressed in moss gametophytes during a desiccation-rehydration cycle and, according to the prior cDNA classification scheme in T. ruralis, are constitutive clones. These findings clearly demonstrated that Rps14, Rps16 and Rpl23 transcripts are retained within the polysomal fractions of desiccated gametophytes. PMID- 11053455 TI - Transport of amino acids (L-valine, L-lysine, L-glutamic acid) and sucrose into plasma membrane vesicles isolated from cotyledons of developing pea seeds. AB - Transport of the amino acids L-valine, L-lysine, and L-glutamic acid and of sucrose was studied in plasma membrane vesicles isolated from developing cotyledons of pea (Pisum sativum L. cv. Marzia). The vesicles were obtained by aqueous polymer two-phase partitioning of a microsomal fraction and the uptake was determined after the imposition of a H(+)-gradient (DeltapH, inside alkaline) and/or an electrical gradient (Deltapsi, inside negative) across the vesicle membrane. In the absence of gradients, a distinct, time-dependent uptake of L valine was measured, which could be enhanced about 2-fold by the imposition of DeltapH. The imposition of Deltapsi stimulated the influx of valine by 20%, both in the absence and in the presence of DeltapH. Uptake of L-lysine was more strongly stimulated by Deltapsi than by DeltapH, and its DeltapH-dependent uptake was enhanced about 6-fold by the simultaneous imposition of Deltapsi. In the absence of gradients the uptake of L-glutamic acid was about 2-fold higher than that of L-valine, but it was not detectably affected by DeltapH or Deltapsi. Although the transport of sucrose was very low, a stimulating effect of DeltapH could be clearly demonstrated. The results lend further support to the contention that during seed development cotyledonary cells employ H(+)-symporters for the active uptake of sucrose and amino acids. PMID- 11053456 TI - A plasma membrane-enriched fraction isolated from the coats of developing pea seeds contains H(+)-symporters for amino acids and sucrose. AB - Aqueous polymer two-phase partitioning was used to obtain a plasma membrane enriched fraction from coats of developing pea (Pisum sativum L.) seeds in the filling stage. Uptake of amino acids and sucrose by vesicles from this fraction was determined after imposition of gradients of proton concentration (DeltapH, inside alkaline) and electrical potential (Deltapsi, inside negative) across the vesicle membrane. The uptake of sucrose and the amino acids L-valine, L-lysine, and L-glutamic acid was stimulated by the imposition of DeltapH. The imposition of Deltapsi, either in the presence or in the absence of DeltapH, stimulated the uptake of L-valine and L-lysine, but had no detectable effect on the uptake of sucrose and L-glutamic acid. The proton-motive-force-driven uptake of all four substrates was abolished by the protonophore carbonylcyanide m-chlorophenyl hydrazone (CCCP). The results demonstrate the presence of H(+)-symporters for sucrose and amino acids in pea seed coats. This is running counter to the previously reported finding that their uptake by isolated pea seed coats was insensitive to CCCP, and that the uptake of sucrose, L-valine, and L-glutamic acid displayed linear kinetics. Possible causes of this discrepancy will be discussed. PMID- 11053457 TI - Imaging spatial and cellular characteristics of low temperature calcium signature after cold acclimation in Arabidopsis. AB - Cooling-induced 'calcium signatures' were imaged in aequorin-expressing Arabidopsis plants after cold acclimation or growth at ambient temperature. In all tissues, signatures were altered after acclimation. Characterization of the components generating this response indicates that cold acclimation increases cold-induced vacuolar Ca(2+) release, but does not affect the influx of extracellular calcium. PMID- 11053458 TI - Rubisco activation state decreases with increasing nitrogen content in apple leaves. AB - Based on the curvilinear relationship between leaf nitrogen content and the initial slope of the response of CO(2) assimilation (A:) to intercellular CO(2) concentrations (C:(i)) in apple, it is hypothesized that Rubisco activation state decreases with increasing leaf N content and this decreased activation state accounts for the curvilinear relationship between leaf N and CO(2) assimilation. A range of leaf N content (1.0-5.0 g m(-2)) was achieved by fertilizing bench grafted Fuji/M.26 apple (Malus domestica Borkh.) trees for 45 d with different N concentrations, using a modified Hoagland's solution. Analysis of A:/C:(i) curves under saturating light indicated that CO(2) assimilation at ambient CO(2) fell within the Rubisco limitation region of the A:/C:(i) curves, regardless of leaf N status. Initial Rubisco activity showed a curvilinear response to leaf N. In contrast, total Rubisco activity increased linearly with increasing leaf N throughout the leaf N range. As a result, Rubisco activation state decreased with increasing leaf N. Both light-saturated CO(2) assimilation at ambient CO(2) and the initial slope of the A:/C:(i) curves were linearly related to initial Rubisco activity, but curvilinearly related to total Rubisco activity. The curvatures in the relationships of both light-saturated CO(2) assimilation at ambient CO(2) and the initial slope of the A:/C:(i) curves with total Rubisco activity were more pronounced than in their relationships with leaf N. This was because the ratio of total Rubisco activity to leaf N increased with increasing leaf N. As leaf N increased, photosynthetic N use efficiency declined with decreasing Rubisco activation state. PMID- 11053459 TI - Rapid recovery of photosystems on rewetting desiccation-tolerant mosses: chlorophyll fluorescence and inhibitor experiments. AB - In the mosses Racomitrium lanuginosum, Anomodon viticulosus and Rhytidiadelphus loreus, after a few days air dry, F:(v)/F:(m) reached, within the first minute of remoistening in the dark, two-thirds or more of the value attained after 40 min. A fast initial phase of recovery was completed within 10-20 min after which further change was slow. Initial recovery of Phi(PSII) in the light was somewhat slower, but was generally substantially complete within a similar time. Remoistening with 0.3 mM cycloheximide (CHX) or 3 mM dithiothreitol (DTT) made little difference to this short-term (40 min) recovery of either F:(v)/F:(m) or Phi(PSII); 3 mM chloramphenicol (CMP) had little effect on recovery of F:(v)/F:(m), but resulted in substantial (though not total) depression of Phi(PSII) and (14)CO(2) uptake. Effects of the protein-synthesis inhibitors and DTT were much more clearly apparent in longer-term experiments (>20 h) but only in the light. In the dark, the three inhibitors had at most only slight effects over periods of 60-100 h. In the light, CMP-treated samples of all three species showed a progressive decline of dark-adapted F:(v)/F:(m), falling to zero within 1-5 d (possibly due to blocking of the turnover of the D1 protein of PSII) and accelerated by DTT. CHX-treated samples showed a similar but slower decline. In the shade-adapted and relatively desiccation-sensitive Rhytidiadelphus loreus, slow recovery of F:(v)/F:(m) continued in the dark even in the presence of CMP and CHX for much of the 142 h of the experiment. The results indicate that in desiccation-tolerant bryophytes recovery of photosynthesis after periods of a few days air dry requires only limited chloroplast protein synthesis and is substantially independent of protein synthesis in the cytoplasm. PMID- 11053460 TI - Architectural and physiological heterogeneity within the synflorescence of the pseudoviviparous grass Poa alpina var. vivipara L. AB - Many biotypes of the northern-hemisphere Arctic-Alpine grass Poa alpina L. reproduce asexually via prolification of the spikelet axis to produce dehiscing shoots. Although capable of photosynthesis, the source-sink characteristics of these synflorescence systems are unknown, including the degree to which plantlets from different regions of the synflorescence are capable of providing for their own carbon requirements, or contributing to other sinks. Photosynthetic rates within the paracladial zone, as determined by infrared gas analysis (IRGA), exceeded respiratory rates by 3-4-fold. (14)CO(2) tracer studies determined that the paracladial zone was not only as efficient at fixing carbon as the youngest fully expanded leaf (per unit dry weight), but that both organs exported carbon mainly basipetally (cf. extensive acropetal export from this leaf in seminiferous grasses). Distal plantlets of the paracladial zone fixed approximately 20% more (14)CO(2) than did proximal plantlets. This was by virtue of their greater dry weight. At dehiscence, 'distal' plantlets were more likely to become established, and possessed relative growth rates more than 10 times those of 'proximal' plantlets. Paracladial heterogeneity was also apparent as an increased proportion of aborted spikelets on proximal paracladia. The possible causes of this heterogeneity are discussed. PMID- 11053461 TI - Marking cell layers with spectinomycin provides a new tool for monitoring cell fate during leaf development. AB - Spectinomycin, an inhibitor of plastid protein synthesis, can be used to mark specific cell layers in the shoot meristem of Brassica napus. Pale yellow-green (YG) plants resulting from spectinomycin-treatment can be propagated indefinitely in vitro. Microscopic examination showed that YG-plants result from inactivation of plastids in the L2 and L3 layers and are composed of a pale green epidermis covering a white mesophyll layer. Epidermal cells of YG and normal green plants are similar and contain 10-20 small pale green plastids. YG plants are equivalent to periclinal chimeras with the important distinction that there is no genotypic difference between the white and green cell layers. Periclinal divisions of epidermal cells take place at all stages of leaf development to produce invaginations of green mesophyll located in sectors of widely varying sizes. A periclinal division rate of 1 in 3000-4000 anticlinal divisions for the adaxial epidermis, was 2-3-fold higher than that estimated for the abaxial epidermis. Analysis of white and green mesophyll showed that chloroplasts are essential for palisade cell differentiation and this requirement is cell-autonomous. Stable marking of cell lineages with spectinomycin is simple, rapid and reveals the requirement for functional plastids in cellular differentiation. PMID- 11053462 TI - Solute balance of a maize (Zea mays L.) source leaf as affected by salt treatment with special emphasis on phloem retranslocation and ion leaching. AB - Strategies for avoiding ion accumulation in leaves of plants grown at high concentration of NaCl (100 mol m(-3)) in the rooting media, i.e. retranslocation via the phloem and leaching from the leaf surface, were quantified for fully developed leaves of maize plants cultivated hydroponically with or without salt, and with or without sprinkling (to induce leaching). Phloem sap, apoplastic fluid, xylem sap, solutes from leaf and root tissues, and the leachate were analysed for carbohydrates, amino acids, malate, and inorganic ions. In spite of a reduced growth rate Na(+) and Cl(-) concentrations in the leaf apoplast remained relatively low (about 4-5 mol m(-3)) under salt treatment. Concentrations of Na(+) and Cl(-) in the phloem sap of salt-treated maize did not exceed 12 and 32 mol m(-3), respectively, and thus remained lower than described for other species. However, phloem transport rates of these ions were higher than reported for other species. The relatively high translocation rate of ions found in maize may be due to the higher carbon translocation rate observed for C(4) plants as opposed to C(3) plants. Approximately 13-36% of the Na(+) and Cl(-) imported into the leaves through the xylem were exported by the phloem. It is concluded that phloem transport plays an important role in controlling the NaCl content of the leaf in maize. Surprisingly, leaching by artificial rain did not affect plant growth. Ion concentrations in the leachate were lower than reported for other plants but increased with NaCl treatment. PMID- 11053463 TI - Effect of different daytime and night-time temperature regimes on the foliar respiration of Pinus taeda: predicting the effect of variable temperature on acclimation. AB - The objectives of this study were to determine the acclimation of loblolly pine (Pinus taeda L.) foliar respiration to different night-time low temperatures, daytime high temperatures, and daily mean temperatures, and then to use the responses of temperature acclimation to various temperature regimes to predict acclimation under fluctuating temperatures. Experiments were conducted on two year-old seedlings in growth chambers using different combinations of day and night-time temperatures. The first experiment exposed trees to 22/22, 29/22, 22/15, and 29/15 degrees C day/night (d/n). When measured at a common temperature (15, 22 or 29 degrees C), respiration rates were lower for trees exposed to higher treatment temperatures and acclimation was influenced by both day and night-time temperature. However, the extent of acclimation did not relate to mean temperature, i.e. respiration rates measured at a common temperature ranked as follows for seedlings exposed to different temperature regimes, 22/15>22/22>29/15 congruent with29/22 degrees C d/n. Rather, acclimation of foliar respiration was linearly related to mean daily respiration rate, where mean daily respiration rate is the average of the respiration rates measured at the day and night-time treatment temperatures. The discrepancy between mean daily respiration rate and mean daily temperature occurred because respiration increased exponentially with increasing temperature. In a second experiment, the same seedlings were exposed to 22/22, 15/15, 25.5/18.5, and 25.5/15 degrees C d/n to test the relationship between mean daily respiration rate and acclimation. As in the first experiment, acclimation was linearly related to mean daily respiration rate. The concept of effective acclimation temperature, which is the temperature at which the mean daily respiration rate occurs, was derived from these results as a means to predict the extent that foliar respiration acclimates to treatment temperature. PMID- 11053464 TI - In situ observation of stomatal movements and gas exchange of Aegopodium podagraria L. in the understorey. AB - Observations of stomata in situ while simultaneously measuring CO(2) gas exchange and transpiration were made in field experiments with Aegopodium podagraria in a highly variable light climate in the understorey of trees. The low background photosynthetic photon flux density (PPFD) caused a slight opening of the stomata and no visible response to sporadic lightflecks. However, if lightflecks were frequent and brighter, slow opening movements were observed. Small apertures were sufficient to allow maximal photosynthetic rates. Therefore, the small apertures observed in low light usually only caused minor stomatal limitations of lightfleck photosynthesis. The response of stomata to step-wise changes in PPFD under different levels of leaf to air vapour pressure difference (Delta(W)) was observed under controlled conditions. High Delta(W) influenced the stomatal response only slightly by reducing stomatal aperture in low light and causing a slight reduction in the initial capacity to utilize high PPFD levels. Under continuous high PPFD, however, stomata opened to the same degree irrespective of Delta(W). Under high Delta(W), opening and closing responses to PPFD-changes were faster, which enabled a rapid removal of the small stomatal limitations of photosynthesis initially present in high Delta(W) after longer periods in low light. It is concluded that A. podagraria maintains a superoptimal aperture in low light which leads to a low instantaneous water use efficiency, but allows an efficient utilization of randomly occurring lightflecks. PMID- 11053465 TI - Quantification of water transport in plants with NMR imaging. AB - A new nuclear magnetic resonance imaging (NMRi) method is described to calculate the characteristics of water transport in plant stems. Here, dynamic NMRi is used as a non-invasive technique to record the distribution of displacements of protons for each pixel in the NMR image. Using the NMR-signal of the stationary water in a reference tube for calibration, the following characteristics can be calculated per pixel without advance knowledge of the flow-profile in that pixel: the amount of stationary water, the amount of flowing water, the cross-sectional area of flow, the average linear flow velocity of the flowing water, and the volume flow. The accuracy of the method is demonstrated with a stem segment of a chrysanthemum flower by comparing the volume flow, measured with NMR, with the actual volumetric uptake, measured with a balance. NMR measurements corresponded to the balance uptake measurements with a rms error of 0.11 mg s(-1) in a range of 0 to 1.8 mg s(-1). Local changes in flow characteristics of individual voxels of a sample (e.g. intact plant) can be studied as a function of time and of any conceivable changes the sample experiences on a time-scale, longer than the measurement time of a complete set of pixel-propagators (17 min). PMID- 11053466 TI - CEF, a sec24 homologue of Arabidopsis thaliana, enhances the survival of yeast under oxidative stress conditions. AB - Budding yeast strains that produced the Arabidopsis thaliana protein CEF or its amino-terminal proline-rich domain were more tolerant to hydroperoxides. CEF is homologous to animal and yeast Sec24 proteins. These data suggest that CEF plays a protective role through protein transport during growth under pro-oxidant conditions. PMID- 11053467 TI - Oxidative stress, heat shock and drought differentially affect expression of a tobacco protein phosphatase 2C. AB - A protein phosphatase 2C (PP2C)-homologous cDNA was isolated from Nicotiana tabacum (NtPP2C1). The deduced protein sequence of 416 amino acids showed the highest degree of similarity to the PP2C of Arabidopsis thaliana (AtPP2CA) implicated in abscisic acid signalling. The expression of NtPP2C1 was strongly induced by drought, but repressed by oxidative stress and heat shock. It is suggested that NtPP2C1 operates at the junction of drought, heat shock and oxidative stress. PMID- 11053469 TI - A comparison of proteins from the developing xylem of compression and non compression wood of branches of sitka spruce (Picea sitchensis) reveals a differentially expressed laccase PMID- 11053468 TI - Characterization of auxin-induced ARRO-1 expression in the primary root of Malus domestica. AB - ARRO-1, a novel 2-oxoacid-dependent dioxygenase (2-ODD) is up-regulated during IBA-induced adventitious root formation in stem discs of Malus domestica. Analysis of ARRO-1's expression profile in the primary root of apple seedlings indicates that it is also highly up-regulated in the root in response to both IAA and IBA, but not 2, 4-D. Auxin-derived evolution of ethylene can be discounted as the source of ARRO-1 induction as ARRO-1 is not induced in the root following treatment with the ethylene precursor ACC. Constitutive expression in the primary root further suggests that ARRO-1's role may be linked to the regulation of natural auxin levels within plant tissues. Significantly, orthologues of ARRO-1 have been identified in Arabidopsis thaliana by means of DNA database analysis which will enable the further molecular characterization of this class of 2-ODD. PMID- 11053470 TI - PTH-Induced downregulation of the type IIa Na/P(i)-cotransporter is independent of known endocytic motifs. AB - Parathyroid hormone (PTH)-induced inhibition of renal proximal tubular Na/P(i) cotransport involves two consecutive steps: endocytosis followed by lysosomal degradation of the type IIa Na/P(i) cotransporter. Tyrosine-, dileucine-, and diacidic-based motifs are suggested to be involved in endocytosis and/or lysosomal targeting of different plasma membrane proteins. The rat type IIa cotransporter (NaPi2) contains two cytoplasmic tyrosine residues (Y) within sequences highly homologous to tyrosine-based motifs (GY(402)FAM and Y(509)RWF), three cytoplasmic dileucine (LL(101), LL(374), and LI(591)) and two cytoplasmic diacidic motifs (EE(81) and EE(616)). We studied the role of these motifs on the PTH-induced retrieval and lysosomal degradation of the NaPi2 cotransporter. To follow its trafficking in vivo, the NaPi2 protein was fused to the carboxyl terminal end of the enhanced green fluorescence protein. This fusion did not impair the apical targeting or the PTH-induced endocytosis of the wild-type cotransporter when transfected in opossum kidney cells. Single and multiple Y and LL mutants retained the apical targeting and the PTH-induced degradation. Mutations of the diacidic motifs were also without effect. These data suggest that the above three motifs are not required for the PTH-induced internalization and/or degradation of the cotransporter. PMID- 11053471 TI - Effect of glucose on intercellular junctions of cultured human peritoneal mesothelial cells. AB - During continuous ambulatory peritoneal dialysis, the peritoneum is directly and continuously exposed to unphysiologic peritoneal dialysis fluid; the resulting mesothelial damage has been suggested to cause loss of ultrafiltration and dialysis efficacy. The present study investigated the effect of a high glucose concentration on cultured human peritoneal mesothelial cells to clarify the cause of decreased dialysis efficacy during prolonged peritoneal dialysis. High glucose caused a concentration-dependent decrease in cell proliferation, damage to the intercellular junctions, and excess production of transforming growth factor-beta (TGF-beta). The levels of intercellular junctional proteins (ZO-1, E-cadherin, and beta-catenin) were decreased, and immuno-staining by anti-ZO-1 and anti- beta catenin antibodies became weaker and often discontinuous along the cell contour. Mannitol had similar but weaker effects at the same osmolality, and an anti-TGF beta neutralizing antibody reduced the effects of high glucose. Therefore, these effects were induced not only by glucose itself but also by hyperosmolality and by a glucose-induced increase of TGF-beta. These findings suggest that the peritoneal mesothelium is damaged by prolonged peritoneal dialysis using high glucose dialysate and that impairment of the intercellular junctions of peritoneal mesothelial cells by high glucose dialysate induces peritoneal hyperpermeability and a progressive reduction in dialysis efficacy. PMID- 11053472 TI - Differential expression of individual UT-A urea transporter isoforms in rat kidney. AB - The rat renal urea transporter UT-A includes four mRNA isoforms: UT-A1, UT-A2, UT A3, and UT-A4. This study detected by rapid amplification of cDNA ends (RACE), primer extension, and ribonuclease protection assay (RPA) a single transcription start site for UT-A1, UT-A3, and UT-A4, distinct from the one for UT-A2 and identified by 3'-RACE new transcripts of UT-A1, UT-A2, and UT-A3, characterized by alternative 3' untranslated sequences (UTR). Expression of an alternative 3'UTR resulted in UT-A1 and UT-A2 transcripts that are approximately 400 bp shorter than the original cDNA. These mRNA isoforms (UT-A1b and UT-A2b) were present in low abundance in the inner medulla. Expression of an alternative 3'UTR for UT-A3 resulted in a 3.5 kb transcript (UT-A3b), which is 1.5 kb longer than the original UT-A3 cDNA. UT-A3b mRNA was easily detected by Northern hybridization in the inner medulla. This study examined whether different states of hydration induce homogeneous changes in mRNA expression of individual UT-A isoforms in the kidney. Analysis of UT-A1, UT-A1b, UT-A2, UT-A2b, UT-A3, and UT A3b mRNA expression in rat kidney revealed that water deprivation markedly increases the relative abundance of UT-A2, UT-A2b, UT-A3, and UT-A3b mRNA in renal inner medulla, whereas UT-A1 and UT-A1b remain almost unchanged. The conclusion is that differential expression of individual UT-A mRNA isoforms occurs in the kidney and probably involves multiple regulatory mechanisms. PMID- 11053473 TI - Localization of inward rectifier potassium channel Kir7.1 in the basolateral membrane of distal nephron and collecting duct. AB - Inward rectifier potassium channels (Kir) play an important role in the K(+) secretion from the kidney. Recently, a new subfamily of Kir, Kir7.1, has been cloned and shown to be present in the kidney as well as in the brain, choroid plexus, thyroid, and intestine. Its cellular and subcellular localization was examined along the renal tubule. Western blot from the kidney cortex showed a single band for Kir7.1 at 52 kD, which was also observed in microdissected segments from the thick ascending limb of Henle, distal convoluted tubule (DCT), connecting tubule, and cortical and medullary collecting ducts. Kir7.1 immunoreactivity was detected predominantly in the DCT, connecting tubule, and cortical collecting duct, with lesser expression in the thick ascending limb of Henle and in the medullary collecting duct. Kir7.1 was detected by electron microscopic immunocytochemistry on the basolateral membrane of the DCT and the principal cells of cortical collecting duct, but neither type A nor type B intercalated cells were stained. The message levels and immunoreactivity were decreased under low-K diet and reversed by low-K diet supplemented with 4% KCl. By the double-labeling immunogold method, both Kir7.1 and Na(+), K(+)-ATPase were independently located on the basolateral membrane. In conclusion, the novel Kir7.1 potassium channel is located predominantly in the basolateral membrane of the distal nephron and collecting duct where it could function together with Na(+), K(+)-ATPase and contribute to cell ion homeostasis and tubular K(+) secretion. PMID- 11053474 TI - Deoxycorticosterone suppresses the effects of losartan in nitric oxide-deficient hypertensive rats. AB - Chronic inhibition of the renin angiotensin system prevents increased BP and renal injury in N(G)-nitro-L-arginine methyl ester (L-NAME) hypertension. However, a relationship between plasma renin activity and the protective effect of chronic angiotensin II (Ang II) blockade has not been established. With this background, this study was undertaken to evaluate how the chronic administration of deoxycortisone acetate (DOCA) modifies the effects of losartan on BP, renal injury, and other variables in L-NAME hypertensive rats. The following groups were used: Control, DOCA, L-NAME, L-NAME + losartan, L-NAME + DOCA, and L-NAME + DOCA + losartan. Tail systolic BP was measured twice a week. After 4-wk evolution, mean arterial pressure and metabolic, morphologic, and renal variables were measured. The final mean arterial pressure values were 116 +/- 6 mmHg for control, 107 +/- 2 mmHg for DOCA, 151 +/- 5 mmHg for L-NAME, 123 +/- 2 mmHg for L NAME + losartan, 170 +/- 3 mmHg for L-NAME + DOCA, and 171 +/- 5.5 mmHg for L NAME + DOCA + losartan. Losartan prevented microalbuminuria, hyaline arteriopathy, and glomerulosclerosis of L-NAME hypertension but was ineffective in L-NAME + DOCA-treated rats. Plasma protein was significantly reduced in the L NAME + DOCA group when compared with control and L-NAME groups, whereas no significant differences were observed in the other groups. Plasma renin activity was suppressed in the DOCA (0.55 +/- 0.2) and L-NAME + DOCA (0.60 +/- 10.2) groups but unsuppressed in the L-NAME + DOCA + losartan group (5.8 +/- 1). The conclusion is that DOCA blocks the preventive effect of losartan on the increased BP and renal injury of L-NAME hypertension, which suggests that DOCA transforms L NAME hypertension into an Ang II-independent model of hypertension. These data also suggest that losartan prevents L-NAME hypertension by blocking the activity of systemic Ang II. PMID- 11053475 TI - Opposing effects of angiotensin II on muscle and renal blood flow under euglycemic conditions. AB - Angiotensin II (Ang II) enhances insulin sensitivity in humans, and this is associated with a paradoxical increase in skeletal muscle blood flow. It is unclear whether these effects are mediated via subtype 1 receptors of Ang II, because these receptors are thought to mediate vasoconstriction. Insulin stimulated glucose uptake (euglycemic clamp technique) and leg muscle blood flow (plethysmography) were measured in nine healthy male volunteers (mean age, 24 +/- 2 yr) on three occasions using a double-blind, placebo-controlled study design. The subjects were allocated in random order to (1) placebo premedication per os plus placebo infusion, (2) placebo premedication per os plus infusion of 5 ng Ang II/kg per min, and (3) premedication with 300 mg of the angiotensin II-1-receptor antagonist irbesartan per os plus infusion of 5 ng Ang II/kg per min. In addition, GFR and effective renal plasma flow were assessed using the steady state inulin- and paraaminohippurate clearance. Insulin sensitivity (i.e., M value) and muscle blood flow after infusion of Ang II (9.3 +/- 1.8 mg/kg per min; 17.7 +/- 2.1 ml/100 g per min) were significantly higher than after placebo infusion (7.2 +/- 1.6 mg/kg per min, P: < 0.02; 13.5 +/- 1.8 ml/100 g per min, P: < 0.01). In contrast, after premedication with irbesartan, they were not significantly different (7.5 +/- 1.7 mg/kg per min; 14.3 +/- 1.9 ml/100 g per min) as compared with placebo infusion. Mean GFR and effective renal plasma flow were significantly lower (P: < 0.01), and renal vascular resistance was significantly higher (P: < 0.01) with Ang II infusion as compared with the placebo infusion study. Premedication with irbesartan almost completely blocked the vasoconstrictive effect of Ang II on renal vasculature. Under hyperinsulinemic euglycemic conditions, infusion of Ang II has opposing effects on regional arterial blood flow, i.e., an increase in skeletal muscle blood flow, but vasoconstriction of renal vasculature. Both effects are antagonized by blockade of subtype 1 Ang II receptors. PMID- 11053476 TI - AP-1 proteins mediate hyperglycemia-induced activation of the human TGF-beta1 promoter in mesangial cells. AB - Hyperglycemia-induced overproduction of the prosclerotic cytokine transforming growth factor-beta1 (TGF-beta1) has been implicated in the pathogenesis of diabetic nephropathy. Because high glucose and phorbol esters (PMA) increase TGF beta1 mRNA levels in mesangial cells, this study was designed to characterize these effects on the human TGF-beta1 promoter activity. With the use of luciferase reporter gene constructs containing TGF-beta1 5'-flanking sequence (from -453 to +11 bp) transfected into mesangial cells, it was found that 30 mM glucose induced a nearly twofold increase in TGF-beta1 promoter activity after 24 h of incubation in human and porcine mesangial cells. Stimulation by PMA was more effective (2.3-fold). Mutagenesis in either one of the two or both activating protein-1 (AP-1) binding sites abolished the high glucose and the PMA effect. Furthermore, addition of the AP-1 inhibitor curcumin obliterated the glucose response. Corresponding experiments revealed that the transcription factor stimulating protein 1 was not involved in mediating the glucose effect. The high glucose-induced TGF-beta1 promoter activation was also prevented by inhibitors of protein kinase C and p38 mitogen-activated proteinkinase. Electrophoretic mobility shift assays with oligonucleotides containing one of the two AP-1 binding sites showed that glucose treatment markedly enhanced the binding activity of nuclear proteins of mesangial cells, particularly to box B. Supershift assays demonstrated that JunD and c-Fos were present in the protein DNA complexes under control and hyperglycemic conditions. The functional and structural results show that glucose regulates human TGF-beta1 gene expression through two adjacent AP-1 binding sites and gives rise to the involvement of protein kinase C and p38 mitogen-activated protein kinase in hyperglycemia induced TGF-beta1 gene expression. PMID- 11053477 TI - Leptospira outer membrane protein activates NF-kappaB and downstream genes expressed in medullary thick ascending limb cells. AB - Tubulointerstitial nephritis is the main manifestation of acute renal damage caused by leptospirosis, but the mechanism remains unexplored. Patients infected with LEPTOSPIRA: shermani in Taiwan disclosed tubular dysfunction particularly in the medullary thick ascending limb of loop of Henle (mTAL), and the related renal damage seems to be underestimated. To elucidate the mechanism of tubular damage, outer membrane protein extract from LEPTOSPIRA: was administered to a model of cultured mTAL cells derived from normal mice. The addition of outer membrane protein extract from L. shermani to cultured mTAL cells induced a significant nuclear DNA binding of the NF-kappa B transcription factor by electrophoresis mobility shift assay. Forty-eight h after adding the outer membrane protein extract (0.2 microg/ml) to the cultured cells, the expression of inducible nitric oxide mRNA increased by 4.2-fold, monocyte chemoattractant protein-1 by 3-fold, and tumor necrosis factor-alpha by 2.4-fold when compared with untreated cells examined by reverse transcription competitive-PCR. Supernatant nitrite, monocyte chemoattractant protein-1, and tumor necrosis factor-alpha protein levels also increased by 1.8-, 7.1-, and 5-fold, respectively. An antiserum raised against L. shermani largely prevented these effects. Outer membrane protein extract from L. bratislava induced fewer effects than L. shermani, and the avirulent nonpathogenic L. biflexa serovar patoc did not induce significant effects in the mTAL cells. In conclusion, L. shermani infection may cause mTAL cell damage and inflammation through the NF-kappa B-associated pathway. Findings of this study may be important in understanding the pathogenesis of tubulointerstitial nephritis caused by these organisms. PMID- 11053478 TI - JB3, an IGF-I receptor antagonist, inhibits early renal growth in diabetic and uninephrectomized rats. AB - Biochemical evidence suggests that insulin-like growth factor I (IGF-I) may play an important role as a mediator of kidney growth. In the present study, an IGF-I receptor antagonist (JB3) was synthesized, and its effect on the renal growth that follows the induction of diabetes or unilateral nephrectomy (UNx) was examined. JB3 was generated by solid phase peptide synthesis. Its activity as an IGF-I antagonist was confirmed in an opossum kidney cell line from its inhibitory effect on the increase in thymidine incorporation into DNA induced by recombinant human IGF-I. Male Wistar rats were anesthetized with halothane and subjected to either the induction of diabetes by streptozotocin (intravenous 60 mg/kg) for 4 d (control animals received citrate buffer) or UNx for 11 d (control animals were sham operated). JB3 was delivered by subcutaneous infusion using an osmotic minipump implanted 3 d before the induction of diabetes or UNx. Kidney wet weight, DNA, and protein all were significantly higher 4 d after the induction of diabetes (24%) or 11 d after UNx (55%). Dose-response studies (1 to 30 microg/kg per day) showed JB3 administration to inhibit the increase in kidney growth in both diabetic and UNx rats. The increase in kidney wet weight, DNA, and protein was significantly lower in UNx rats that were treated with JB3 10 microg/kg per day (P: < 0.05) than in saline vehicle controls but was abolished in diabetic rats that were treated with JB3 3 microg/kg per day (P: < 0. 01). Increasing the dose of JB3 to 30 microg/kg per day was associated with a decrease in its inhibitory effect, resulting in bell-shaped dose-response curves. JB3 administration had no effect on the blood glucose concentration or food consumption by either diabetic or nondiabetic animals. The results support the concept of IGF-I as an important mediator of the early renal growth that follows the induction of diabetes or UNx in the rat. PMID- 11053479 TI - Glomerular monocyte-macrophage features in ANCA-positive renal vasculitis and cryoglobulinemic nephritis. AB - Although it is widely known that many macrophages are present in glomeruli of antineutrophil cytoplasmic antibody (ANCA)-positive renal vasculitis (ANCA + RV) and are believed to contribute to necrotizing extracapillary damage, their precise role is not yet completely understood, especially in humans. The goal of this study was to provide evidence of glomerular macrophage properties in human vasculitis. Twenty-five renal biopsies of ANCA + RV and 18 cases of cryoglobulinemic glomerulonephritis (cryoGN), a disease characterized by massive glomerular macrophage infiltration but absence of necrotizing extracapillary lesions, were selected, and macrophage number, adhesion, acute activation, proliferation, and apoptosis were analyzed by immunohistochemistry and in situ hybridization. Accumulation of macrophages in ANCA + RV was found in areas of glomerular active lesions, whereas in cryoGN, they homogeneously occupied the entire glomerular tuft. Considering the areas of accumulation, comparable macrophage numbers were detected in both diseases. Glomerular vascular cell adhesion molecule-1 was found only in ANCA + RV and only in areas of active lesions. Acute macrophage activation (HLA class II, 27E10) and proinflammatory cytokine production (tumor necrosis factor-alpha, interleukin-1alpha) were prominent in ANCA + RV, whereas in cryoGN, 30% of glomerular macrophages seemed activated and cytokine expression was limited to a few glomerular cells (P: = 0.01). Moreover, only in ANCA + RV proliferative markers were shown on glomerular macrophages and apoptotic macrophages were found. From the data, it seems that ANCA + RV and cryoGN differ profoundly in macrophage properties, namely adhesion, proliferation, and apoptotic clearance. Moreover, acute activation and cytokine production seem to be present in a greater number of macrophages in ANCA + RV, giving this disease a stronger severity that could be taken into account for therapeutic strategies. PMID- 11053480 TI - Interleukin-17 and CD40-ligand synergistically enhance cytokine and chemokine production by renal epithelial cells. AB - Renal allograft rejection is characterized by an influx of inflammatory cells. Interaction between infiltrating T cells and resident parenchymal cells might play an important role in the ongoing inflammatory response. The present study demonstrates that CD40L, a product of activated T cells, is locally expressed in kidneys undergoing rejection. Furthermore, during rejection, CD40 expression not only is present on most graft infiltrating cells but also is increased on resident tubular epithelial cells (TEC). To obtain more detailed insight in the consequences of T cell/TEC interaction, we analyzed the production of chemokines, including interleukin-8 (IL-8), monocyte chemoattractant protein-1 (MCP-1) and regulated upon activation, normal T cell expressed and secreted (RANTES), and the production of IL-6 by cultured human primary TEC in response to activation with CD40L in vitro. In addition, we studied the interaction with IL-17, a T-cell specific cytokine previously demonstrated to be present during renal allograft rejection. The results, obtained by enzyme-linked immunosorbent assay, indicate that simultaneous activation of TEC with IL-17 and CD40L synergistically enhances production of IL-6 (2.1-fold higher than sum of single stimulations) and the chemokines IL-8 (15-fold) and RANTES (5.8-fold) as demonstrated by statistical analysis (P: < 0.05), whereas effects on MCP-1 (1.4-fold) are additive. Part of the synergy can be explained by increased CD40 expression on TEC upon IL-17 stimulation. The synergy is not unique for TEC, because similar responses were found with human synoviocytes and a foreskin fibroblast cell line (FS4). Stimulation of TEC with CD40L results in activation of NF-kappaB and induction of cytokine production by IL-17 and CD40L is prevented by addition of the NF-kappaB inhibitor pyrrolidine dithiocarbamate. These data suggest an important role for T cells in renal allograft rejection by acting on parenchymal cells via both soluble mediators and direct cellular contact. PMID- 11053481 TI - Rats transgenic for human renin and human angiotensinogen as a model for gestational hypertension. AB - Animal models of gestational hypertension are problematic. A novel mouse model was described earlier. The dams in that study were transgenic for human angiotensinogen and the sires for human renin; human renin was expressed in and produced by the placenta. This model was adapted to the rat, which has greater utility in terms of chronic instrumentation and physiologic measurements. Female rats transgenic for human angiotensinogen were mated with rats transgenic for human renin. Telemetry BP increased on day 5 of pregnancy from 110/80 mmHg to as high as 180/140 mmHg, while heart rate increased slightly. The renin transgene was expressed in the placenta, which resulted in increased human plasma renin concentration from 0 to 937 +/- 800 ng angiotensin I ml/h; the values returned to 0 after delivery. Female rats transgenic for human renin that were mated with male rats transgenic for human angiotensinogen in contrast exhibited a decrease in BP. In these rats, human angiotensinogen in plasma remained undetectable. Double transgenic offspring of these transgenic rats developed hypertension and end-organ damage, regardless of the source of the transgenes. The conclusion is that transgenic rats that bear human renin and angiotensinogen genes make an attractive model for gestational hypertension. The rat model will have greater utility than the mouse model. PMID- 11053482 TI - Polymorphism of angiotensin converting enzyme, angiotensinogen, and angiotensin II type 1 receptor genes and end-stage renal failure in IgA nephropathy: IGARAS- a study of 274 Men. AB - The impact of renin-angiotensin system (RAS) gene polymorphism on the prognosis of IgA nephropathy (IgAN) is still debated. A longitudinal study of renal prognosis in patients with IgAN was conducted to search retrospectively for a genotype-phenotype association between RAS polymorphisms and end-stage renal failure (ESRF). A classification based on serum creatinine (S(cr)) and 24-h proteinuria (24-P) measured at the time of renal biopsy was used to estimate the risk of ESRF in IgAN: stage 1 (S(cr) 150 micromol/L and 24-P < 1 g or S(cr) < or = 150 micromol/L and 24-P > or = 1 g), stage 3 (S(cr) > 150 micromol/L and 24-P > or = 1 g). Deletion/insertion polymorphism (D/I) of the angiotensin I converting enzyme gene, M235T polymorphism (T/M) of the angiotensinogen gene and A1166C polymorphism (C/A) of the angiotensin II type 1 receptor gene were determined in 274 Caucasian men with biopsy-proven IgAN (n = 86, 112, and 76 in stages 1, 2, and 3, respectively). Mean global follow-up was 6 +/- 5 yr after renal biopsy. For stages 1, 2, and 3, ESRF developed in 7 (8. 1%), 39 (34.8%), and 49 (64.4%) cases (P: < 0.0001), 11.7 +/- 4, 5.4 +/- 4, and 2 +/- 2 yr, respectively, after renal biopsy (P: < 0.001). The distributions of the three genotypes into the three stages were similar. Different distributions were observed when patients were grouped by stage and genotype: ID+DD: 72% in stage 1 versus 84.6% in stages 2 + 3 (P: = 0.02; kappa = 0.14); MT+TT: 66.2% in stages 1 + 2 versus 78.9% in stage 3 (P: = 0.04; kappa = 0.09); and AA+AC: 89.9% in stages 1 + 2 versus 97.4% in stage 3 (P: = 0.04; kappa = -0.1). However, with the use of the Cox proportional hazard model, none of the three genotypes was found to have predictive value for renal survival. Compared with S(cr) and 24-P, genotypes DD, TT, and AA are unlikely to serve as clinically useful predictors of ESRF in IgAN. PMID- 11053483 TI - Evidence of gene-gene interactions in the genetic susceptibility to renal impairment after unilateral nephrectomy. AB - The number of patients with hypertension-associated end-stage renal failure (ESRF) continues to increase despite improved antihypertensive management and early detection programs. Variation for the development of renal complications in hypertension may reflect independent genetic susceptibility to ESRF. The genetically hypertensive fawn-hooded rat is characterized by the early presence of systolic hypertension, glomerular hypertension, progressive proteinuria (UPV), and focal glomerulosclerosis (FGS), resulting in premature death as a result of renal failure. In the present study, the genetic basis of hypertension-associated ESRF in an F2 intercross consisting of 337 animals, in which systolic BP, UPV, albuminuria, and FGS, were studied at 8 wk after a unilateral nephrectomy performed at 5 to 6 wk of age. A total genome scan, consisting of 418 markers, was used to identify regions that contribute to the pathogenesis of UPV and FGS. Linkage analysis revealed five loci involved in the development of renal impairment. Of these five, two (Rf-1, Rf-2) had been identified previously. There seems to be strong interactive effects between the various loci and their impact on UPV and the other parameters of renal impairment, as well as an interaction with BP. In particular, Rf-1 seems to play a major role in determining the severity of the disease. This study is the first to report the interaction of more than two loci to produce progressive renal failure, suggesting that the genetic dissection of renal failure in humans will require understanding of how multiple genes interact with each other and BP to produce ESRF. PMID- 11053484 TI - Renal epithelium is a previously unrecognized site of HIV-1 infection. AB - The striking emergence of an epidemic of HIV-related renal disease in patients with end-stage renal disease provided the rationale for the exploration of whether HIV-1 directly infects renal parenchymal cells. Renal glomerular and tubular epithelial cells contain HIV-1 mRNA and DNA, indicating infection by HIV 1. In addition, circularized viral DNA, a marker of recent nuclear import of full length, reverse-transcribed RNA, was detected in the biopsies, suggesting active replication in renal tissue. Infiltrating infected leukocytes harbored more viral mRNA than renal epithelium. Identification of this novel reservoir suggests that effectively targeting the kidney with antiretrovirals may be critical for patients who are seropositive with renal disease. Thus, renal epithelium constitutes a unique and previously unrecognized cell target for HIV-1 infection. PMID- 11053485 TI - Calcemic activity of 19-Nor-1,25(OH)(2)D(2) decreases with duration of treatment. AB - 19-Nor-1,25(OH)(2)D(2) (19-norD(2)) has been shown to suppress parathyroid hormone effectively, but with lower calcemic activity than 1,25(OH)(2)D(3). The present study investigated potential mechanisms to explain the reduced calcemic response to 19-norD(2). Tissue localization of [(3)H]19-norD(2) or[(3)H]1,25(OH)(2)D(3) after a single injection was not different. Intestinal calcium absorption and bone mobilization, measured in vitamin D-deficient rats 24 h after single injections of 60 or 600 pmol of 19-norD(2) or 1,25(OH)(2)D(3), were enhanced to a similar degree by the two compounds. However, when normal rats were treated every other day with 240 pmol of 19-norD(2) or 1,25(OH)(2)D(3), increases in serum calcium were identical 24 h after the first injection but diverged thereafter with significantly lower serum calcium in the 19-norD(2) treated rats by 5 d. Intestinal calcium absorption and bone calcium mobilization were reassessed in vitamin D-deficient rats after seven daily injections of 600 pmol of 19-norD(2) or 1, 25(OH)(2)D(3), and both parameters were significantly lower in the 19-norD(2)-treated rats. Pharmacokinetic analysis after seven daily injections of 600 pmol of 19-norD(2) or 1,25(OH)(2)D(3) showed similar localization to the intestine and bone. In addition, intestinal vitamin D receptor levels were not different after 1 wk of treatment with 19-norD(2) or 1,25(OH)(2)D(3). In conclusion, the low calcemic activity of 19-norD(2) seems to be due to an acquired, postreceptor resistance of the intestine and bone to chronic treatment with the analog. PMID- 11053486 TI - Glomerular permselectivity at the onset of nephropathy in type 2 diabetes mellitus. AB - The development of microalbuminuria in individuals with type 2 diabetes mellitus is associated with a 10-fold increase in the risk of progression to overt nephropathy and eventual end-stage renal failure. The present study reports long term (up to 8 yr) follow-up of 43 Pima Indians with type 2 diabetes detected on screening to have microalbuminuria. The natural history of albuminuria in these individuals included progression to overt proteinuria (urinary albumin excretion > or = 300 mg/d) in half of the participants by 7 yr of follow-up. The size selectivity of the glomerular barrier was also investigated using dextran sieving and pore theory. Whereas a comparison group of macroalbuminuric Pima Indians had an excess of large pores that served as a macromolecular "shunt," individuals with microalbuminuria had a shunt size no different from long-term diabetic, normoalbuminuric control subjects. An abrupt transition from little or no relationship to a highly significant and positive relationship between increasing albuminuria and shunt size occurred at an albumin-to-creatinine ratio of approximately 3000 mg/g. Shunt size in macroalbuminuric individuals correlated with the extent of foot process broadening. Podocyte foot processes in microalbuminuric participants were not different from those in control subjects. In conclusion, although microalbuminuria in type 2 diabetic Pima Indians often heralds progressive glomerular injury, it is not the result of defects in the size permselectivity of the glomerular barrier but rather of changes in either glomerular charge selectivity or tubular handling of filtered proteins or of a combination of these two factors. PMID- 11053487 TI - Effects of growth hormone on leptin metabolism and energy expenditure in hemodialysis patients with protein-calorie malnutrition. AB - The relationships among growth hormone (GH), leptin, and resting energy expenditure (REE) are not understood. It has been reported that in malnourished hemodialysis patients, GH increases muscle protein synthesis, a process that requires energy. The present study evaluated the arterial levels and the forearm exchange of leptin, as well as the REE of the same patients during their participation in the same study, in four sequential 6-wk periods: I, baseline; II, GH treatment; III, washout; and IV, GH + intradialytic parenteral nutrition. During periods II and IV, patients received GH (5 mg three times per week). REE rose by 5% in period II, declined during period III, and rose by 7% during period IV. Basal leptin levels were low (2.0 +/- 0.19 ng/L). Insulin and leptin levels, as well as leptin release from the forearm, were unchanged during periods I through III but rose (+ 36%; P: < 0.05) during period IV. Changes in arterial leptin were directly related to changes in forearm leptin release (P: < 0.002), indicating a role of leptin production by peripheral tissues on leptinemia. Changes in leptin release were directly related to insulin (P: < 0.001) and, less consistently, to insulin-like growth factor-binding protein-1 levels (P: < 0.02). Similarly, variations in leptin levels were directly related to insulin (P: < 0.01). Variations in REE were not related to variations in leptin or insulin levels but to changes in muscle protein synthesis (P: < 0.025). The data show that in malnourished hemodialysis patients, treatment with GH is not invariably associated with an increase in leptin production. An increase in leptin release by peripheral tissues and leptin levels occurs only in the setting of hyperinsulinemia. The increase in REE that is induced by treatment with GH is not dependent on changes in leptin but is largely accounted for by the energy cost of the stimulation of muscle protein synthesis. PMID- 11053488 TI - Interleukin-1 receptor antagonist synthesis by peripheral blood mononuclear cells: a novel predictor of morbidity among hemodialysis patients. AB - Proinflammatory cytokines have been implicated in the short- and long-term morbidity experienced by hemodialysis (HD) patients. The present study, which is based on long-term follow-up of a cohort of 37 patients, relates peripheral blood mononuclear cell (PBMC) interleukin-1 receptor antagonist (IL-1Ra) synthesis (a reliable marker of IL-1beta synthesis in HD patients) and plasma levels of an acute phase reactant, lipopolysaccharide binding protein (LBP), to clinical outcomes. In July 1993, predialysis blood samples from these patients were collected and IL-1Ra synthesis by PBMC and plasma LBP was measured. Hospital records were reviewed and patient follow-up data were obtained until December 1997 (54 mo) or death, whichever occurred earlier. The effect of age, diabetes, endotoxin- and IgG-stimulated IL-1Ra synthesis, and plasma LBP levels on mortality was assessed using the Cox proportional hazard regression model. Poisson regression was used to determine potential relationships between the number of outcome events and each continuous risk factor. Twenty-two patients (59%) died during the follow-up period. Mortality was unrelated to IL-1Ra synthesis but did increase with age (relative risk, 1.05/yr; P: = 0.01) and diabetes (relative risk, 3.00/yr; P: = 0.03). Cardiovascular event rates were higher among older individuals and in those with higher endotoxin-stimulated PBMC IL-1Ra synthesis. Cardiovascular events increased with plasma LBP levels in the range of 9,000 to 12,000 pg/ml but then seemed to decrease. In contrast, older age and low IgG-stimulated IL-1Ra synthesis were associated with an increased risk of infectious events. The results of this study demonstrate an interesting link between stimulus-dependent variability in IL-1Ra synthesis by PBMC and clinical outcomes among patients on chronic HD and provide interesting targets for therapeutic interventions in this vulnerable patient population. PMID- 11053489 TI - Tunneled-cuffed catheter associated infections in hemodialysis patients who are seropositive for the human immunodeficiency virus. AB - Infection rates in tunneled-cuffed catheters (TCC) are reported to be higher in immunocompromised patients. The purpose of this study was to evaluate TCC associated infection rates in patients with HIV infection (HIV+). Data were collected in 40 HIV + patients and 41 controls (C), and in 118 TCC (HIV+, 58; C, 60) for 28,146 catheter days (HIV+, 16,227; C, 11,919). There were no significant differences in the TCC bacteremia rates (HIV+, 2.23 versus C, 2.53 per 1000 TCC days, P: = NS) or in the TCC exit site infection rates (HIV+, 2.20 versus C, 2.24 per 1000 TCC days, P: = NS) between the groups. The number of TCC removed due to infection was also similar, (HIV+ versus C: 17 versus 15%, P: = NS). In the HIV+ group, the association of hepatitis B surface antigenemia with TCC exit site infection was dependent on the history of injection drug use. Black race was a significant risk factor for higher TCC exit site infection rates, whereas prophylactic antibiotic use and high CD4 count were significantly associated with lower TCC exit site infection rates. None of the factors significantly predicted bacteremia rate in either group (HIV+ or C). In comparison to controls, HIV+ patients had a fivefold increased risk of having a Gramnegative organism (P: = 0.02) and a sevenfold increased risk of a fungal isolate (P: = 0.08), although the latter finding was not statistically significant. HIV infection is not a significant risk factor for TCC-associated infection but is associated with a higher prevalence of Gram-negative and fungal species. PMID- 11053490 TI - Detection of Na(+) transporter proteins in urine. AB - Previous studies have established that the vasopressin-regulated water channel of the collecting duct, aquaporin-2, is excreted in the urine, providing a means for assessment of regulation and dysregulation of aquaporin-2 in humans. This article addresses the hypothesis that membrane transporters from upstream nephron segments are normally detectable in urine. The experiments employed rabbit polyclonal antibodies against the major Na transporters of the proximal tubule (the type 3 Na-H exchanger [NHE3]), the thick ascending limb of Henle's loop (the bumetanide-sensitive Na-K-2Cl cotransporter [NKCC2]), and the distal convoluted tubule (the thiazide-sensitive Na-Cl cotransporter [NCC]) in immunoblotting experiments. All three of these transporters were readily detectable as high molecular weight complexes present in lowdensity membrane fractions from urine of normal rats. Cross linking studies of NHE3, NKCC2, and NCC revealed that high molecular weight complexes are normally present in renal tissue. The molecular weights of the complexes in urine matched those of the cross-linked complexes in native kidney tissue. The presence in urine of integral membrane proteins representative of each nephron segment raises the possibility that limited or comprehensive proteomic analysis of urine samples may be useful in clinical settings. PMID- 11053491 TI - Desensitization of soluble guanylate cyclase in renal cortex during endotoxemia in mice. AB - Acute endotoxemic renal failure involves renal vasoconstriction, which presumably occurs despite increased nitric oxide (NO) generation by inducible NO synthase in the kidney. The present study examined the hypothesis that the renal vasoconstriction during endotoxemia occurs in part because of desensitization of soluble guanylate cyclase (sGC). Endotoxic shock was induced in male B6/129F2/J mice by an intraperitoneal injection of Escherichia coli lipopolysaccharide. The endotoxemia resulted in shock and renal failure as evidenced by a decrease in mean arterial pressure and an increase in serum creatinine and urea nitrogen. Serum NO increased in a time-dependent manner, reaching the highest levels at 24 h, in parallel with induction of inducible NO synthase protein in the renal cortex. In renal cortical slices obtained from endotoxemic mice, cyclic guanosine monophosphate (cGMP) increased significantly at 6 h and 15 h as compared with control but normalized at 24 h after injection of lipopolysaccharide. Incubation of renal cortical slices in the presence of a phosphodiesterase inhibitor isobutylmethylxantine did not alter the pattern of changes in cGMP. Incubation of renal cortical slices with 2 mM sodium nitroprusside resulted in a similar accumulation of cGMP in slices taken from control and endotoxemic mice at 6 h and 15 h. However, in slices from 24-h endotoxemic mice, accumulation of cGMP in response to sodium nitroprusside was significantly lower. This lower stimulability of sGC was not paralleled by a decrease in its abundance in renal cortex on immunoblot. Taken together, these results demonstrate a desensitization of sGC in renal cortex during endotoxemia, which may contribute to the associated renal vasoconstriction. PMID- 11053492 TI - HIV: much at stake with kidneys? PMID- 11053493 TI - The immune tolerance network: the "Holy Grail" comes to the clinic. PMID- 11053494 TI - Malarial acute renal failure. PMID- 11053495 TI - Endogenous production of fixed acid and the measurement of the net balance of acid in normal subjects. PMID- 11053496 TI - The biology of somatotropin in adipose tissue growth and nutrient partitioning. AB - During the past 20 years, much has been learned about how porcine somatotropin (pST) affects growth and nutrient partitioning in growing pigs. The development of techniques to produce large quantities of recombinantly derived pST enabled numerous long-term studies to be conducted in which the effects of daily pST administration could be evaluated. Collectively, these studies established that treatment of growing pigs with pST markedly stimulated muscle growth and, concurrently, reduced fat deposition. In growing pigs, maximally effective doses of pST increase average daily gain as much as 10-20%, improve feed efficiency 15 30%, decrease adipose tissue mass and lipid accretion rates by as much as 50-80% and concurrently increase protein deposition by 50%. These effects are associated with a decrease in feed intake of approximately 10-15%. These responses occur because pST has a wide array of biological effects that modulate nutrient partitioning between adipose tissue and skeletal muscle. The decrease in adipose tissue growth is due to a reduction in lipogenesis that is the consequence of pST blunting the effects of many insulin-dependent events. With respect to fatty acid synthase (FAS), a pace-setting enzyme in the lipogenic pathway, enzyme activity is markedly reduced by pST. This is the result of a pST-mediated decrease in FAS mRNA levels that occurs because FAS gene transcription is decreased. The consequence of the decrease in lipid synthesis is that adipocyte hypertrophy is impaired and, hence, tissue growth. This review will provide an overview of some of the biological effects of pST in adipose tissue and will discuss what is known about the underlying mechanisms that account for these effects. PMID- 11053497 TI - Arginine nutrition and cardiovascular function. AB - L-Arginine (Arg) is the substrate for the synthesis of nitric oxide (NO), the endothelium-derived relaxing factor essential for regulating vascular tone and hemodynamics. NO stimulates angiogenesis, but inhibits endothelin-1 release, leukocyte adhesion, platelet aggregation, superoxide generation, the expression of vascular cell adhesion molecules and monocyte chemotactic peptides, and smooth muscle cell proliferation. Arg exerts its vascular actions also through NO independent effects, including membrane depolarization, syntheses of creatine, proline and polyamines, secretion of insulin, growth hormone, glucagon and prolactin, plasmin generation and fibrinogenolysis, superoxide scavenging and inhibition of leukocyte adhesion to nonendothelial matrix. Compelling evidence shows that enteral or parenteral administration of Arg reverses endothelial dysfunction associated with major cardiovascular risk factors (hypercholesterolemia, smoking, hypertension, diabetes, obesity/insulin resistance and aging) and ameliorates many common cardiovascular disorders (coronary and peripheral arterial disease, ischemia/reperfusion injury, and heart failure). Dietary Arg supplementation may represent a potentially novel nutritional strategy for preventing and treating cardiovascular disease. PMID- 11053498 TI - Stimulation of in vitro rat muscle protein synthesis by leucine decreases with age. AB - Aging is characterized by a decrease of muscle mass associated with a decrease in postprandial anabolism. This study was performed to gain a better understanding of the intracellular mechanisms involved in the stimulation of muscle protein synthesis by amino acids and their role in the decrease of muscle sensitivity to food intake during aging. The effects of amino acids or leucine alone were assessed in vitro on epitrochlearis muscle from young, adult and old rats. Protein synthesis was assessed by incorporation of radiolabeled phenylalanine into protein and p70 S6 kinase activity by incorporation of (32)P into a synthetic substrate. Amino acids, at physiologic concentrations, stimulated muscle protein synthesis (P < 0.05) and leucine reproduced this effect. The intracellular targets of amino acids were phosphatidylinositol 3' kinase and the rapamycin-sensitive pathways mammalian target of rapamycin (mTOR)/p70 S6 kinase. In old rats, the sensitivity of muscle protein synthesis to leucine was lower than in adults (P < 0.05) and this paralleled the lesser ability of leucine to stimulate the rapamycin-sensitive pathways (P < 0.05). We demonstrated that amino acids and leucine stimulate muscle protein synthesis and that aging is associated with a decrease in this effect. However, because aged rats are still able to respond normally to high leucine concentrations, we hypothesize that a nutritional manipulation increasing the availability of this amino acid to muscle could be beneficial in maintaining the postprandial stimulation of protein synthesis. PMID- 11053499 TI - Medium- and long-chain fatty acids differentially modulate interleukin-8 secretion in human fetal intestinal epithelial cells. AB - The primary therapeutic effects of enteral nutrition in patients with Crohn's disease have been reported previously. Although the quantity and type of fat in enteral nutrition are considered to be important, it is unclear how fat modulates mucosal inflammatory responses in the intestine. In the present study, we evaluated the effects of medium-chain and long-chain fatty acids (MCFA and LCFA) on interleukin (IL)-8 secretion in a fetal intestinal epithelial cell line, intestine-407 cells. IL-8 expression was evaluated at the protein and mRNA levels. The activation of nuclear factor-kappaB was assessed with an electrophoretic gel mobility shift assay. The addition of oleic acid (LCFA) micelles, but not octanoic acid (MCFA) micelles, weakly but significantly enhanced basal IL-8 secretion in the intestine-407 cells. The addition of MCFA (5 mmol/L) induced a 40% increase in IL-1beta-induced IL-8 secretion and a 35% increase in tumor necrosis factor (TNF)-alpha-induced IL-8 secretion, respectively. The addition of LCFA (5 mmol/L) induced a 140% increase in IL-1beta induced IL-8 secretion and a 110% increase in TNF-alpha-induced IL-8 secretion, respectively. These responses were also observed at the mRNA levels. The electrophoretic gel mobility shift assay indicated that both MCFA and LCFA enhanced IL-1beta- and TNF-alpha-induced nuclear factor-kappaB activation. We demonstrated the proinflammatory activities of MCFA and especially LCFA. It is likely that medium-chain triglycerides may be more suitable than long-chain triglycerides as an energy source in enteral diets in the treatment of patients with Crohn's disease. PMID- 11053500 TI - Soy protein isolate reduces the oxidizability of LDL and the generation of oxidized LDL autoantibodies in rabbits with diet-induced atherosclerosis. AB - The incidence of atherosclerosis can be modified by diet, and plant-derived proteins have a beneficial effect, but the underlying mechanisms remain unclear. It has been suggested that oxidized LDL (oxLDL) and autoantibodies against oxLDL are important in the development of atherosclerosis. We analyzed these factors in rabbits fed a nonpurified diet supplemented with high cholesterol (10.0 g/kg) containing either 270.0 g/kg casein (CAS, n = 10) or 270.0 g/kg soy protein isolate (SPI, n = 10) for 2 mo. Plasma and purified serum LDL from rabbits were analyzed at d 0, 15, 30, 45 and 60 of treatment, and the size of atherosclerotic lesions was evaluated at d 60 of treatment. CAS-fed rabbits had significantly higher plasma cholesterol at d 15-45 and LDL cholesterol levels at d 15 and 30. Levels of trilinolein and phosphatidylcholine hydroperoxides were higher in the LDL fraction of rabbits fed CAS than in those fed SPI. Also, CAS-fed rabbits had higher levels of highly oxidized LDL [monoclonal antibody (mAb) 24-reactive oxLDL] in plasma at d 60, whereas SPI-fed rabbits had higher levels of minimally oxidized LDL (mAb 28-reactive oxLDL) at d 45. These results were consistent with the earlier formation of anti-oxLDL antibodies and the presence of a larger area of atherosclerotic lesion in rabbits fed the CAS diet. These data indicate the importance of both the type of dietary protein used in the induction of atherosclerosis and the relevance of immunologic mechanisms in addition to biochemical and physiologic factors in the pathogenesis of atherosclerosis. PMID- 11053501 TI - 1,25-Dihydroxycholecalciferol prevents and ameliorates symptoms of experimental murine inflammatory bowel disease. AB - Anecdotal data suggest that the amount of vitamin D available in the environment either from sunshine exposure or diet may be an important factor affecting the development of inflammatory bowel disease (IBD) in humans. We tested the vitamin D hypothesis in an experimental animal model of IBD. Interleukin (IL)-10 knockout (KO) mice, which spontaneously develop symptoms resembling human IBD, were made vitamin D deficient, vitamin D sufficient or supplemented with active vitamin D (1,25-dihydroxycholecalciferol). Vitamin D-deficient IL-10 KO mice rapidly developed diarrhea and a wasting disease, which induced mortality. In contrast, vitamin D-sufficient IL-10 KO mice did not develop diarrhea, waste or die. Supplementation with 50 IU of cholecalciferol (5.0 microgram/d) or 1, 25 dihydroxycholecalciferol (0.005 microgram/d) significantly (P < 0. 05) ameliorated symptoms of IBD in IL-10 KO mice. 1, 25-Dihydroxycholecalciferol treatment (0.2 microgram/d) for as little as 2 wk blocked the progression and ameliorated (P < 0.05) symptoms in IL-10 KO mice with already established IBD. PMID- 11053502 TI - Fluctuations in dietary methionine intake do not alter plasma homocysteine concentration in healthy men. AB - A moderate elevation in plasma total homocysteine (tHcy) has been established as an independent risk factor for vascular disease. An important exogenous source of homocysteine is methionine found in foods rich in animal protein. We investigated the response of tHcy to fluctuations in methionine intake in a cross-over intervention trial (two arms). Healthy men (n = 52; 19-29 y) were screened for habitual methionine intake using a food-frequency questionnaire. Subjects in the top quartile for methionine intake (n = 13), with a baseline fasting tHcy of 7.01 +/- 1.84 micromol/L (mean +/- SD), were randomly assigned to receive either a low methionine intervention diet for 1 wk followed by a control diet for 1 wk or vice versa. Simultaneously, those in the bottom quartile for methionine intake (n = 11), with a fasting plasma tHcy of 9.79 +/- 7. 20 micromol/L (mean +/- SD), received either a high methionine intervention diet for 1 wk followed by a control diet or vice-versa. All subjects had serum folate, red-cell folate, serum vitamin B-12 and plasma pyridoxal phosphate (PLP) concentrations within normal ranges. During the intervention, subjects in the top quartile for methionine intake reduced their daily methionine intake 79%, from 1969 +/- 639 to 407 +/- 83 mg/d (P: 28 wk), symptoms of nausea, faintness and night blindness were reduced with vitamin A, but not beta-carotene, supplementation. Vitamin A supplementation shortened the length of labor by 1.5 h 50 min among nulliparous and multiparous women, respectively. Both interventions reduced the postpartum prevalence of at least four loose stools and night blindness. beta-Carotene supplementation also reduced symptoms of high fever postpartum. The mean number of days of any reported illness symptoms was 3-4 per wk throughout pregnancy. Among women receiving vitamin A, the total number of days of illness symptoms accrued over the last 12 wk of pregnancy was lower by 5 d compared with the placebo recipients. We found the burden of pregnancy-related illness symptoms to be high in this rural area of Nepal where antenatal care is poor and most deliveries occur at home. Maternal vitamin A or beta-carotene supplementation resulted in a reduction in the prevalence of selected illness symptoms during late pregnancy, at the time of birth and during 6 mo postpartum, suggesting that a diet adequate in vitamin A may be important for improving women's reproductive health. PMID- 11053507 TI - Premature complementary feeding is associated with poorer growth of vietnamese children. AB - The objective of this longitudinal study was to investigate the association between the premature initiation of complementary feeding and physical growth of children. Four cohorts of newborn children were included, consisting of 90 infants born in 1981, 90 in 1982, 60 infants in 1983 and 60 in 1984. The weights and heights of children were measured monthly up to 1 y, then every 3 mo for y 2 and 3, and once every 6 mo in y 4. Information on feeding practices and diseases of the children was obtained by interviewing the mothers at each home visit. All but three children (98.6%) were breast-fed. Although 87.1% of the mothers breast fed their children for at least 1 y, only 3.3% of the infants were breast-fed exclusively at the age of 4 mo. In the analyses of growth, care was taken to address the biases of reverse causality, regression to the mean and confounding. There was little association between feeding pattern at 15 d and growth in length in mo 1. However, partially breast-fed and weaned infants gained weight more slowly than those exclusively or predominantly breast-fed. From 1 to 3 mo, exclusively breast-fed infants grew more quickly in both weight and length, followed by predominantly breast-fed infants. From 3 to 6 mo, exclusively breast fed infants gained more weight compared with the other groups, but there was a slight difference (P = 0.047) in length gain only between exclusively and partially breast-fed infants. In the older period (6-12 mo), exclusively and predominantly breast-fed infants grew in length more quickly than partially breast-fed and weaned groups. However, there was no difference in weight gain among groups. Morbidity from diarrhea and acute respiratory infections was significantly lower for the >/=3 mo exclusively breast-fed group (chi(2) and Fisher-Exact Test). Over nearly the whole age range from 1 mo to 4 y, Z-scores for all indices (weight-for-age, height-for-age and weight-for-height) of the children who received complementary food were significantly lower than those of children who were exclusively breast-fed for at least 3 mo (repeated measures ANOVA, adjusted for sex, family size, maternal education and family income). These results show a long-term deterioration of physical growth in infants who received premature complementary feeding and confirm the importance of exclusive breast-feeding for infants for at least 3 mo. PMID- 11053508 TI - Supplemental vitamin A improves anemia and growth in anemic school children in Tanzania. AB - We conducted a randomized controlled trial of the effects of dietary supplements on anemia, weight and height in 136 anemic school children from a low socioeconomic background in Bagamoyo District schools in Tanzania. The aim of the current study was to investigate the impact of dietary supplements on anemia and anthropometric indices of anemic school children. The supplements were vitamin A alone, iron and vitamin A, iron alone or placebo, administered in a double blinded design for 3 mo. All supplements were provided with local corn meals. Hemoglobin concentration, body weight and height were measured at baseline and at follow-up after supplementation. Vitamin A supplementation increased the mean hemoglobin concentration by 13.5 g/L compared with 3.5 g/L for placebo [P < 0.0001, 95% confidence interval (CI) 6.19-13.57), the mean body weight by 0.6 kg compared with 0.2 kg for placebo (P < 0.0001, 95% CI 0.19-0.65) and the mean height by 0.4 cm compared with 0.1 cm for placebo (P = 0.0009, 95% CI 0.08-0.42). However, the group of children who received combined vitamin A and iron supplementation had the greatest improvements in all indicators compared with placebo (18.5 g/L, P < 0.0001, 95% CI 14.81-22.23; 0.7 kg, P < 0. 0001, 95% CI 0.43-0.88 and 0.4 cm, P < 0.0001, 95% CI 0.22-0.56 for hemoglobin, weight and height, respectively). It is likely that vitamin A supplementation may have a useful role in combating the problems of vitamin A deficiency and anemia, as well as in improving children's growth, in developing countries. PMID- 11053509 TI - Daily iron supplementation is more effective than twice weekly iron supplementation in pregnant women in Pakistan in a randomized double-blind clinical trial. AB - In the context of limited effectiveness of iron supplementation programs, intermittent iron supplementation is currently under debate as a possible alternative strategy that may enhance the effectiveness of operational programs. This field-based trial assessed the outcome of twice weekly iron supplementation compared to daily in Pakistan. A double-blind, randomized, clinical trial was conducted in Northern Pakistan. Anemic pregnant women (n = 191) were assigned to receive daily (200 mg ferrous sulfate) or twice weekly (2 x 200 mg ferrous sulfate) iron supplementation. Hemoglobin was measured at baseline and at 4-wk intervals for up to 12 wk. Serum ferritin was measured at baseline and 8 or 12 wk. Analysis was by intention to treat. The two groups did not differ in age, parity, sociodemographic characteristics, hemoglobin or serum ferritin concentrations at baseline. Women who received iron daily had a greater rise in hemoglobin compared with women who received iron twice weekly (17.8 +/- 1.8 vs. 3.8 +/- 1.2 g/L, P < 0.001). The serum ferritin concentrations increased by 17.7 +/- 3.9 microgram/L (P < 0.001) in the daily supplemented group and did not change in the twice weekly group. Daily iron supplementation remained superior to twice weekly supplementation after controlling initial hemoglobin Z-scores and duration of treatment. The body mass index (BMI) modified the effect of daily versus twice weekly iron supplementation. For every unit increase in BMI, the difference between the two treatment groups was reduced by 0.0014 (final hemoglobin Z-score; P = 0.027). We recommend continuation of daily iron supplementation as opposed to intermittent iron supplementation in pregnant women in developing countries. PMID- 11053510 TI - Anemia and deficiencies of folate and vitamin B-6 are common and vary with season in Chinese women of childbearing age. AB - Little is known about the micronutrient status of Chinese women of childbearing age. We assessed nonfasting plasma concentrations of folic acid, vitamin B-12, vitamin B-6 (as pyridoxal-5'-phosphate), hemoglobin (Hb), ferritin and transferrin receptor (TfR) in 563 nonpregnant textile workers aged 21-34 y from Anqing, China. All women had obtained permission to become pregnant and were participating in a prospective study of pregnancy outcomes. Mean (SD) plasma concentrations were 9.7 (4.1) nmol/L folic acid, 367 (128) pmol/L vitamin B-12, 40.2 (15.8) nmol/L vitamin B-6, 108 (12. 9) g/L Hb, 42.6 (34.2) microgram/L ferritin and 5.2 (2.7) mg/L TfR. Twenty-three percent of women had biochemical evidence of folic acid deficiency, 26% were deficient in vitamin B-6 and 10% had low vitamin B-12. Overall, 44% of women were deficient in at least one B vitamin. Although anemia (Hb < 120 g/L) was detected in 80% of women, only 17% had depleted iron stores (ferritin < 12 microgram/L); 11% had elevated TfR concentrations. Distinct seasonal trends were observed in the prevalence of moderate anemia (Hb < 100 g/L) and deficiencies of folic acid and vitamin B-6, with significantly lower concentrations of folate and Hb occurring in summer and lower concentrations of vitamin B-6 occurring in winter and spring than in other seasons. We conclude that deficiencies of folic acid, vitamin B-6 and iron were relatively common in this sample of Chinese women of childbearing age and were contributing to the high prevalence of anemia. Without appropriate supplementation, these deficiencies could jeopardize the women's health and increase their risk of adverse pregnancy outcomes. PMID- 11053511 TI - Food stamps are associated with food security and dietary intake of inner-city preschoolers from Hartford, Connecticut. AB - The goal of the present study was to examine the association of the Food Stamp Program with the food security and dietary intake of low-income children from Hartford, CT, who were enrolled in the Supplemental Food Program for Women, Infants, and Children (WIC). We compared the food and nutrition situation of low income preschoolers who received food stamps (FS, n = 59) with that of those who did not receive food stamps (NFS, n = 40). Children were an average age of 2.7 +/ 0.6 y, and 95% were receiving WIC benefits at the time of the study. Groups were comparable in demographic characteristics, but the socioeconomic status of the FS group was lower than that of the NFS group (P < 0.05). Food security was assessed with the Radimer/Cornell hunger scale, and dietary intake was assessed with a single 24-h recall and a 14-item food frequency questionnaire. Multivariate analyses within the FS group indicated that a monthly duration of food stamps of <4 wk was a predictor of household food security (odds ratio 0.10, 95% confidence interval 0.02-0.56). Food stamp use was associated with above-median energy adjusted intakes of vitamin B-6 (3.13, 1.16-8.45), folate (2.92, 1.09-7.81) and iron (3.72, 1.31-10.54). The NFS children were more likely to consume <8 mg iron/d (3.73, 1.09-12.80). These results suggest that the Food Stamp Program is associated with food security and preschoolers' micronutrient intake. PMID- 11053512 TI - Biomarkers of human colonic cell growth are influenced differently by a history of colonic neoplasia and the consumption of acarbose. AB - The nutritional effects of butyrate on the colonic mucosa and studies of transformed cells suggest that butyrate has anti-colon cancer effects. If butyrate has antineoplastic effects, mucosal growth contrasts between normal subjects and those with a history of colonic neoplasia would parallel changes in growth characteristics caused by butyrate in a colon neoplasia population. To test this hypothesis, rectal biopsies from a survey of colonoscopy patients (n = 50) with and without a history of colonic neoplasia (controls) were compared. Similarly, rectal biopsies were compared from subjects (n = 44) with a colon neoplasia history in an acarbose-placebo crossover trial. Control subjects in the colonoscopy survey had higher bromodeoxyuridine (BrdU) uptake than subjects with a history of neoplasia (P = 0.05). The control subjects also had a higher correlation of BrdU and Ki-67 labeling (P = 0.003). Both findings were paralleled by acarbose use. Acarbose augmented BrdU uptake (P = 0.0001) and improved the correlation of BrdU and Ki-67 labeling (P = 0.013). Acarbose also augmented fecal butyrate (P = 0.0001), which was positively correlated with Ki-67 labeling (P = 0.003). p52 antigen had an earlier pattern of crypt distribution in subjects with a history of colon neoplasia but was not affected by acarbose use. Lewis-Y antigen was expressed earlier in the crypt with acarbose but had similar expression in the colonoscopy survey groups. The use of acarbose to enhance fecal butyrate concentration produced mucosal changes paralleling the findings in control subjects as opposed to those with neoplasia, supporting the concept of an antineoplastic role for butyrate. PMID- 11053513 TI - Prouroguanylin overproduction and localization in the intestine of zinc-deficient rats. AB - Identification of the upregulation of preprouroguanylin mRNA in the rat small intestine during zinc deficiency provides a potential mechanistic link between production of the intestinal hormone uroguanylin and the diarrhea that may accompany zinc deficiency. In the current study, in situ hybridization demonstrated that the number of preprouroguanylin mRNA-expressing cells was significantly higher in zinc-deficient rats than in zinc-adequate rats. Immunohistochemical studies, with a uroguanylin peptide affinity-purified antibody, demonstrated that immunoreactivity was localized to the tips of villi of the duodenum and jejunum in zinc-adequate rats. However, positive cells were scattered throughout the villus of zinc-deficient rats. A subset of cells, perhaps enterochromaffin cells, exhibited the predominant staining, whereas no specific staining was found in goblet cells or lymphocytes of the lamina propria. Western blotting demonstrated that the expression of prouroguanylin in both duodenum and jejunum was elevated by dietary zinc depletion. These results show that dietary zinc deficiency upregulates prouroguanylin in intestinal cells, which is consistent with a role for uroguanylin in the etiology of diarrhea observed in human zinc deficiency. PMID- 11053514 TI - Polymeric proanthocyanidins are catabolized by human colonic microflora into low molecular-weight phenolic acids. AB - Polymeric proanthocyanidins are common constituents of many foods and beverages. Their fate in the human body remains largely unknown. Their metabolism by human colonic microflora incubated in vitro in anoxic conditions has been investigated using nonlabeled and (14)C-labeled purified proanthocyanidin polymers. Polymers were almost totally degraded after 48 h of incubation. Phenylacetic, phenylpropionic and phenylvaleric acids, monohydroxylated mainly in the meta or para position, were identified as metabolites by gas chromatography coupled to mass spectrometry (GC-MS). Yields were similar to those previously reported for flavonoid monomers. These results provide the first evidence of degradation of dietary phenolic polymers into low-molecular-weight aromatic compounds. To understand the nutritional properties of proanthocyanidins, it is therefore essential to consider the biological properties of these metabolites. PMID- 11053515 TI - Rat jejunum controls luminal thiol-disulfide redox. AB - The control of luminal thiol-disulfide redox state may be important for several intestinal functions, including absorption of iron or selenium and maintenance of mucus fluidity. Disulfides are present in the diet, and although luminal thiols are supplied in bile, little is known about the ability of the small intestine to reduce disulfides to maintain the luminal thiol-disulfide redox state. The objective of the current study was to determine whether the isolated, vascularly perfused jejunum, free from biliary thiols, could reduce intraluminal glutathione disulfide (GSSG) to glutathione (GSH). GSSG was introduced in a deoxygenated solution to inhibit the reoxidation of any GSH formed, and preparations were pretreated with acivicin to inhibit the degradation of GSH by gamma glutamyltransferase. GSSG (250 micromol/L) was reduced to GSH, with the luminal redox potential (E(h)) for GSSG/2GSH changing from >0 to -111, -132 and -143 mV at 10, 20 and 30 min, respectively. The E(h) for luminal cystine/2cysteine was approximately 20 mV more reducing than that for GSSG/2GSH at each time point, suggesting that cysteine could function in the reduction of GSSG in the lumen. Measurements in specific regions showed that GSSG reduction was more rapid in the duodenum and proximal jejunum than in the distal jejunum. Preparations without acivicin treatment showed that E(h) values were unaffected by inhibition of gamma glutamyltransferase despite differences in GSH and cysteine pool sizes. Rat intestine has a mechanism to adjust the luminal thiol-disulfide redox. In principle, dysfunction of this mechanism could contribute to malabsorption or other nutritional disorders. PMID- 11053516 TI - Temporal response of hepatic threonine dehydrogenase in chickens to the initial consumption of a threonine-imbalanced diet. AB - Amino acid imbalances contribute to higher requirements of amino acids than would occur if the dietary profile of amino acids perfectly matched the requirements. The mechanisms of imbalances have not been fully elucidated. Because threonine dehydrogenase (TDH) activity in liver mitochondria increases in chicks and rats subjected to threonine imbalance, the current study was carried out to determine whether the change in TDH activity occurs rapidly enough after the consumption of an imbalanced diet to be considered a possible primary metabolic response. In a series of experiments, Leghorn chicks were allowed free access to a semipurified basal diet marginally limited in threonine or the same diet containing a mixture of indispensable amino acids (IAA) lacking threonine to cause a threonine imbalance. In the first experiment, dietary supplements of 5.5 and 11.1% IAA were used to determine a level of supplement that would cause a robust response in the specific activity of TDH. Feed intake, body weight gains and efficiency of feed utilization were lower and specific activities of TDH were higher in chicks fed 11.1% IAA than in those fed 5.5% IAA. In subsequent experiments, hepatic TDH activities and plasma amino acid profiles of the control and experimental groups were determined at 1. 5, 3, 6, 12 and 24 h after the first offering of the diet containing 11.1% IAA. The specific activities of TDH in chicks fed the IAA supplement were 40-150% higher (P < 0.05) and plasma threonine concentrations were 42-53% lower (P < 0.05) than in chicks fed the basal diet at all times except 1.5 h. These results indicate that changes in the capacity for threonine degradation via TDH may occur in the liver within a few hours after the consumption of a threonine-imbalanced diet and suggest the possibility that altered TDH activity may contribute to the increased threonine requirement associated with threonine imbalance. PMID- 11053517 TI - Cholesterol reduction by glucomannan and chitosan is mediated by changes in cholesterol absorption and bile acid and fat excretion in rats. AB - Glucomannan, a viscous polysaccharide, and chitosan, a derivative of chitin, have both been demonstrated to lower cholesterol in animals. However, the mechanism of cholesterol lowering has not been established for either material. This study was conducted to determine the effect of glucomannan (G), chitosan (CH), or an equal mixture of the two (G + CH) on cholesterol absorption and fat and bile acid excretion. Rats were fed a modified AIN-93G diet for 18 d containing 0.125 g/100 g cholesterol and initially 10 g/100 g of the test materials or cellulose (C) as the control. However, the concentration of test materials and cellulose was reduced to 7.5 g/100 g after 1 wk due to lower weight gain compared with controls. Total liver cholesterol was significantly reduced in G, CH and G + CH groups compared with the C group. The intestinal contents supernatant viscosity of the C and the CH groups was negligible, whereas both G and G + CH produced high viscosities. Cholesterol absorption, measured by the fecal isotope ratio method, was significantly reduced from 37.5% in the C group to 20.2% in G, 18.2% in G + CH and 9.4% in CH. Daily fecal fat excretion did not differ between the C and G groups, but was significantly greater in G + CH and CH compared with the C and G groups. Daily fecal bile acid excretion was significantly greater in the CH and G + CH groups compared with the C and G groups. These results suggest that G lowered liver cholesterol by a viscosity-mediated interference of cholesterol absorption. In contrast, CH appears to lower cholesterol through a different mechanism. PMID- 11053518 TI - Platycodi radix affects lipid metabolism in mice with high fat diet-induced obesity. AB - An aqueous extract of Platycodi radix inhibited the hydrolysis of triolein emulsified with phosphatidylcholine by pancreatic lipase in vitro and it reduced the elevation of rat plasma triacylglycerol level 2-4 h after oral administration of a lipid emulsion containing corn oil. These preliminary results suggested that the aqueous extract of Platycodi radix may inhibit the intestinal absorption of dietary fat by inhibiting its hydrolysis. Therefore, we examined the antiobesity activity of the aqueous extract of Platycodi radix by testing whether the extract prevented the obesity induced by feeding a high fat diet to mice for 8 wk. Body weights at 3-8 wk and the final parametrial adipose tissue weights were significantly lower in mice fed the high fat diet containing 5% aqueous extract of Platycodi radix than in the controls fed the high fat diet. The aqueous extract of Platycodi radix also significantly reduced hepatic triacylglycerol concentrations that were elevated in mice fed the high fat diet alone. Inulin, which is a major component of Platycodi radix, had no effect on the hydrolysis of triolein emulsified with phosphatidylcholine by pancreatic lipase in vitro, and did not prevent obesity or the fatty liver induced by the high fat diet. On the other hand, the total saponin fraction of the aqueous extract inhibited pancreatic lipase activity in vitro. Therefore, the antiobesity effect of the aqueous extract of Platycodi radix in mice fed a high fat diet may be due in part to the inhibition of intestinal absorption of dietary fat by the saponins of Platycodi radix. PMID- 11053519 TI - Intestinal transport of quercetin glycosides in rats involves both deglycosylation and interaction with the hexose transport pathway. AB - Flavonoids are polyphenolic plant secondary metabolites with antioxidant and other biological activities potentially beneficial to health. Food-borne flavonoids occur mainly as glycosides, some of which can be absorbed in the human small intestine; however, the mechanism of uptake is uncertain. We used isolated preparations of rat small intestine to compare the uptake of the quercetin aglycone with that of some quercetin glucosides commonly found in foods, and investigated interactions between quercetin-3-glucoside and the intestinal hexose transport pathway. The nature of any metabolism of quercetin and its glucosides during small intestinal transport in vitro was determined by HPLC. The presence of quercetin-3-glucoside in the mucosal medium suppressed the uptake of labeled galactose by competitive inhibition and stimulated the efflux of preloaded galactose. Quercetin-3-glucoside and quercetin-4'-glucoside, but not quercetin 3,4'-diglucoside, were transported into everted sacs significantly more quickly than quercetin aglycone. Intact quercetin glucosides were not detected in mucosal tissue or within the serosal compartment, but both free quercetin and its metabolites were present, mainly as quercetin-3-glucuronide and quercetin-7 glucuronide. Evidently, quercetin derived from quercetin-3-glucoside passes across the small intestinal epithelium more rapidly than free quercetin aglycone. Monoglucosides of quercetin interact with the sodium-dependent glucose transporter. During passage across the epithelium, quercetin-3-glucoside is rapidly deglycosylated and then glucuronidated. PMID- 11053520 TI - Transport mechanisms of the imino acid L-proline in the human intestinal epithelial caco-2 cell line. AB - The intestinal transport of L-proline (L-Pro) has been investigated in various animal species with the use of different tissue preparations. Because major qualitative differences have been observed among the species, it is difficult to extent the results obtained with animal models to humans. In addition, studies on human tissue are lacking because of difficulties in obtaining material for experiments. To characterize the mechanisms involved in the intestinal absorption of L-Pro in humans, the transport of this nonessential imino acid was studied in monolayers of human intestinal Caco-2 cells that were cultivated on microporous membranes. In this model, L-Pro was transported selectively in the apical (AP)-to basolateral (BL) direction. This transport was significantly reduced by metabolic inhibitors and by an incubation at 4 degrees C; it was Na(+) dependent and showed competition with (methylamino)-alpha-isobutyric acid and L-hydroxyproline. By contrast, transport in the BL-to-AP direction resulted to a large extent from passive movement (paracellular passage and transcellular diffusion). L-Pro accumulation by Caco-2 cells was significantly greater from the AP pole than from the BL pole. About 30-50% of the accumulated molecules were incorporated into newly synthesized proteins in a process inhibited by cycloheximide, whereas the remainder were extensively metabolized into non-amino acid compounds. L-Pro accumulations from the AP and BL poles were both Na(+) dependent, but they exhibited different characteristics. AP accumulation was inhibited by competition with (methylamino)-alpha-isobutyric acid, L-hydroxyproline and, to a lesser extent, D-Pro, whereas BL accumulation was inhibited by competition with L hydroxyproline, (methylamino)-alpha-isobutyric acid, alpha-aminoisobutyric acid, L-histidine and small neutral amino acids. The results indicate that AP-to-BL transport and AP accumulation of L-Pro exhibited very different characteristics than BL-to-AP transport and BL accumulation. PMID- 11053521 TI - Transport mechanisms of the large neutral amino acid L-phenylalanine in the human intestinal epithelial caco-2 cell line. AB - The transepithelial transport and the intracellular accumulation of the large neutral amino acid L-phenylalanine (L-Phe) were studied in monolayers of Caco-2 cells, cultivated in a bicameral insert system, to characterize the mechanisms involved in the absorption of this essential amino acid by the human intestinal mucosa. In our model, L-Phe was transported selectively in the apical (AP)-to basolateral (BL) direction. AP-to-BL transport of L-Phe was temperature dependent and Na(+) independent, increased in the absence of protein synthesis and showed competition with large neutral and cationic amino acids. By contrast, transport in the BL-to-AP direction mainly resulted from passive movement (probably paracellular passage and transcellular diffusion). L-Phe accumulation into Caco-2 cells was higher from the BL pole than from the AP pole and characterized by the incorporation of most of the accumulated molecules into newly synthesized proteins. In addition, L-Phe accumulation was Na(+) dependent from both poles, whereas only accumulation from the AP pole was sensitive to inhibition by both large neutral and cationic amino acids. These results suggest that the processes involved in AP-to-BL transport and AP accumulation of this amino acid are very different from those involved in BL-to-AP transport and BL accumulation. PMID- 11053522 TI - Amount of dietary fat and type of soluble fiber independently modulate postabsorptive conversion of beta-carotene to vitamin A in mongolian gerbils. AB - Current dietary guidelines recommend a decrease in fat intake and an increase in fiber consumption. Decreased bioavailability (BV) of carotenoids is thought to be associated with both of these recommendations. A 2 x 4 factorial design was used to test the effects of dietary fat level at 10 or 30% of total energy and fiber type using no fiber, silica, citrus pectin or oat gum (7 g/100 g) on beta carotene (betaC) BV in 4- to 5-wk-old Mongolian gerbils. We assessed BV as both accumulation of betaC and bioconversion of betaC to vitamin A (VA) in tissues. A VA- and betaC-deficient diet was fed for 1 wk followed by one of eight isocaloric, semipurified diets supplemented with carrot powder [ approximately 1 microgram betaC, 0.5 microgram alpha-carotene (alphaC)/kJ diet] for 2 wk (n = 12/group). Increasing dietary fat resulted in higher VA (P: = 0.074) and lower betaC (P: = 0.0001) stores in the liver, suggesting that consumption of high fat diets enhances conversion of betaC to VA. The effect of soluble fiber on hepatic VA storage was dependent on fiber type. Consumption of citrus pectin resulted in lower hepatic VA stores and higher hepatic betaC stores compared with all other groups, suggesting less conversion of betaC to VA. In contrast, consumption of oat gum resulted in hepatic VA and betaC stores that were higher (P = 0.012) and lower (P = 0.022), respectively, than those of citrus pectin-fed gerbils. The level of dietary fat consumed with soluble fiber had no interactive effects on hepatic VA, betaC or alphaC stores. Results demonstrate that betaC BV is independently affected by dietary fat level and type of soluble fiber, and suggest that these dietary components modulate postabsorptive conversion of betaC to VA. This study confirms the negative effects of citrus pectin on betaC BV, and suggests that oat gum does not adversely affect betaC BV. PMID- 11053523 TI - Glucose is absorbed in a sodium-dependent manner from forestomach contents of sheep. AB - Intraruminal glucose is thought to be completely converted to short-chain fatty acids (SCFA) by symbiotic microorganisms. Nevertheless, earlier in vitro studies evidenced the expression of the sodium glucose-linked transporter (SGLT)-1, in the ovine ruminal epithelium. The present study aimed to determine whether the ruminal SGLT-1 is functionally important in vivo. In a first experimental series using the emptied, washed, and isolated reticulorumen of sheep, 6.3% of glucose was absorbed from an intraruminal buffer solution (2 L, 128 mmol/L Na(+), 0.5 mmol/L glucose, 0 mmol/L galactose) within 30 min (P < 0.001). Reducing Na(+) concentration to 10 mmol/L resulted in complete inhibition of glucose absorption, and the addition of 10 mmol/L galactose (at 128 mmol/L Na(+)) induced a small but insignificant inhibition. In a second experimental series, the addition of 12 mmol/L glucose to an initially glucose-free buffer led to an increase in the transruminal potential difference from 34.4 to 37.1 mV within 4 min (P < 0.001). From the 12 mmol/L glucose-containing buffer, 11.0% of glucose was absorbed within 30 min (P < 0.05). In all experiments, microbial glucose degradation in the reticulorumen was prevented by adding cefuroxime (100 mg/L) and colistin methanesulfonate (25 mg/L) to the buffer solution. The effectiveness of antimicrobial treatment was verified by ex vivo incubations of buffer samples drawn from the reticulorumen. We conclude that glucose is absorbed in a sodium dependent manner from the reticulorumen at low and high glucose concentrations. Absorption at high glucose concentrations is of nutritional importance because it counteracts the genesis of ruminal lactic acidosis. PMID- 11053524 TI - Structured lipids improve fat absorption in normal and malabsorbing rats. AB - The presence of medium-chain fatty acids in dietary fatty acid as well as the triacylglycerol structure may influence the absorption and lymphatic transport of fatty acids. We compared the lymphatic transport and recovery of fatty acids from four intragastrically administered fats based on rapeseed oil and decanoic acid in two rat models of normal absorption and malabsorption, respectively. The fats were: 1) a fat with a regiospecific structure, 2) a similar fat but with a random distribution of fatty acids in the triacylglycerol molecule, 3) a physical mixture of tridecanoin and rapeseed oil and 4) rapeseed oil as control. Lymph samples were collected for 24 h. Significantly higher recoveries were observed of total fatty acids, oleic acid, linoleic acid and linolenic acid from the specific oil in malabsorbing rats and of linoleic acid in normal rats fed specific oil compared with those fed rapeseed oil. Furthermore, the recoveries of oleic acid and linolenic acid from the specific oil in normal rats were higher than those from the other oils. In malabsorbing rats, the transport of all fats was approximately 90% less than that of normal rats. The present study demonstrates improved hydrolysis and absorption of the specific oil compared with the other oils examined both in rats with normal absorption and in rats with malabsorption. PMID- 11053525 TI - Free amino acids are absorbed faster and assimilated more efficiently than protein in postlarval Senegal sole (Solea senegalensis). AB - To improve the formulation of diets for the early stages of marine fish, assimilation rates of free amino acids (FAA) and protein in postlarval Senegal sole (Solea senegalensis) were determined. Fish (2.45 +/- 0.87 mg dry weight) were tube fed 36 nL of a diet of FAA containing L-[(35)S] methionine (FAA diet) or bovine serum albumin, containing L-[methylated-(14)C]bovine serum albumin (Prot-diet), both at a concentration of 4.08 g/L. A time series was performed, and the amounts of label in incubation water, liver, gut and body carcass were quantified. The FAA diet was absorbed with a 3.5-times-higher transfer rate (P < 0.001) from the gut into the larval body tissues compared with the Prot-diet. The FAA diet also was assimilated with greater efficiency than the Prot-diet (80% versus 58%, P: = 0.001). If we assume that the label present in the gut represents amino acids incorporated into the intestinal tissue, the assimilation efficiencies for the two diets were 89 and 64%. Therefore, FAA seems to be superior to protein as a dietary source of amino acids in Senegal sole postlarvae. However, because the absorption dynamics of protein and FAA differ, care should be taken when using the sources together to avoid amino acid imbalance. PMID- 11053526 TI - Leptin rapidly inhibits hypothalamic neuropeptide Y secretion and stimulates corticotropin-releasing hormone secretion in adrenalectomized mice. AB - Leptin may rapidly inhibit food intake by altering the secretion of hypothalamic neuropeptides such as neuropeptide Y (NPY), a stimulator of food intake, and/or corticotropin-releasing hormone (CRH), an inhibitor of food intake. We measured concentrations of NPY and CRH in specific hypothalamic regions [i.e., arcuate nucleus (ARC), paraventricular nucleus (PVN), ventromedial nucleus and dorsomedial nucleus] of 7- to 8-wk-old lean and ob/ob mice at 1 or 3 h after intracerebroventricular leptin administration. No rapid-onset effects of leptin on hypothalamic NPY or CRH concentrations were observed in intact mice. The addition of leptin to hypothalamic preparations from intact mice also did not alter NPY or CRH secretion. Glucocorticoids may oppose leptin actions. Consistent with this, leptin administration to adrenalectomized mice markedly reduced CRH concentrations in the ARC within 3 h after injection. This rapid reduction in CRH concentration in the ARC after leptin administration is more likely due to stimulated CRH release from this region than to decreased synthesis/transport from the PVN because leptin stimulates CRH synthesis in the PVN. Within 20 min after exposure to leptin, NPY secretion from hypothalamic preparations obtained from adrenalectomized mice was lowered by 27% and CRH secretion was elevated by 51%. The current study demonstrates that leptin rapidly influences the secretion of hypothalamic NPY and CRH and that these actions of leptin within the hypothalamus are restrained by the presence of endogenous corticosterone. PMID- 11053527 TI - Chronic marginal iron intakes during early development in mice result in persistent changes in dopamine metabolism and myelin composition. AB - Marginal iron (Fe) deficiency is prevalent in children worldwide, yet the behavioral and biochemical effects of chronic marginal Fe intakes during early development are not well characterized. Using a murine model, previous work in our laboratory demonstrated persistent behavioral disturbances as a consequence of marginal Fe intakes during early development. In the present study, Swiss Webster mice fed a control Fe diet (75 microgram Fe/g diet, n = 13 litters) or marginal Fe diet (14 microgram Fe/g diet, n = 16 litters) during gestation and through postnatal day (PND) 75 were killed on PND 75 for assessment of tissue mineral concentrations, dopamine metabolism, myelin fatty acid composition, and c and m-aconitase activities. In addition, these outcomes were assessed in a group of offspring (n = 13 litters) fed a marginal Fe diet during gestation and lactation and then fed a control diet from PND 21-75. Marginal Fe mice demonstrated significant differences in brain iron concentrations, dopamine metabolism and myelin fatty acid composition relative to control mice; however, no difference in c- or m-aconitase activity was demonstrated in the brain. The postnatal consumption of Fe-adequate diets among marginal Fe offspring did not fully reverse all of the observed biochemical disturbances. This study demonstrates that chronic marginal Fe intakes during early development can result in significant changes in brain biochemistry. The persistence of some of these biochemical changes after postnatal Fe supplementation suggests that they are an irreversible consequence of developmental Fe restriction. PMID- 11053528 TI - Iron deficiency alters dopamine transporter functioning in rat striatum. AB - Iron deficiency anemia in early life produces profound changes in both in vivo and in vitro evaluations of dopamine (DA) functioning. This study employed both behavioral and biochemical approaches to examine the biological bases of alterations in striatal DA metabolism seen in iron-deficient rats. The purpose was to determine whether the DA transporter (DAT) was functionally altered in postweaning iron deficiency. Male and female 21-d-old Sprague-Dawley rats (n = 40) were fed either an iron-deficient (ID) diet (3 mg Fe/kg diet) or a control (CN) diet (35 mg Fe/kg diet) for 4 wk before behavioral testing. Motor activity responses to graded doses (3.75-30 mg/kg body) of the DA uptake inhibitor, cocaine, were significantly blunted in iron-deficient rats with a 50% higher half maximal effective dose (ED(50)) in both males and females (CN-female, 7.1 +/- 0.9 mg/kg; ID-female, 11.2 +/-1.3 mg/kg; CN-male, 12.0 +/- 0.7 mg/kg; and ID-male, 17.0 +/- 1.8 mg/kg). Radioligand binding assays with (3)H-1-(2-(diphenylmethoxy) ethyl)-4-(3-phenylpropyl) piperazine ((3)H-GBR12935) demonstrated that iron deficiency did not alter the affinity of the ligand for the DAT but did significantly decrease the density of the transporter by 30% in caudate putamen and 20% in nucleus accumbens. Iron deficiency also significantly decreased (3)H DA uptake into striatal synaptosomes, but did not affect release of DA with potassium chloride stimulation. These experiments provide supporting evidence that elevated levels of extracellular DA in the striatum of iron-deficient rats is likely to be the result of decreased DAT functioning and not increased rates of release. PMID- 11053529 TI - Zinc and copper intakes and their major food sources for older adults in the 1994 96 continuing survey of food intakes by individuals (CSFII). AB - Zinc and copper are two trace minerals essential for important biochemical functions and necessary for maintaining health throughout life. Several national food surveys revealed marginally to moderately low contents of both nutrients in the typical American diet. Using data from the respondents >/= 60 y old in the 1994-96 Continuing Survey of Food Intakes by Individuals (CSFII), we examined average dietary intakes of zinc, copper and relevant dietary factors; primary dietary contributors of zinc and copper; and Zn:Cu ratios of the primary dietary contributors. Data were analyzed with the use of a chi(2) test, Student's t test and multivariate analysis of covariance with Bonferroni correction. The daily zinc intake was 12 +/- 6.4 mg for men and 8.0 +/- 4.0 mg for women (P < 0.05); the daily copper intake was 1.3 +/- 0.7 mg for men and 1.0 +/- 0.5 mg for women (P < 0.05). Foods such as beef, ground beef, legumes, poultry, ready-to-eat and hot cereals, and pork constituted the major sources of zinc. Copper consumption was contributed mainly by legumes, potato and potato products, nuts and seeds, and beef. The less-than-recommended intakes of zinc and copper by the elderly were likely associated with age, low income and less education. The intakes of zinc and copper could be improved by more frequent consumption of food sources rich in these minerals. An inherent limitation of this study was the use of the 24-h dietary recall method, which may underestimate usual dietary intakes. Nonetheless, this study affirms the need for assessment of zinc and copper nutriture in the elderly. PMID- 11053530 TI - Vitamin A status assessment in rats with (13)C(4)-retinyl acetate and gas chromatography/combustion/isotope ratio mass spectrometry. AB - Vitamin A assessment methods that indirectly determine liver reserves are still in development. The deuterated vitamin A assay has been successfully applied in several population groups, but large doses of vitamin A must be used and the gas chromatography/mass spectrometry analysis is not very sensitive. Therefore, 10,11,14,15-(13)C(4)-retinyl acetate was synthesized using a modified Wittig Horner procedure. Thereafter, female Sprague-Dawley rats (n = 47) were fed a vitamin A-deficient diet and divided into three groups: low (L), moderate (M) and high (H) vitamin A. Groups L, M and H were supplemented with 35, 70 and 350 nmol of unlabeled retinyl acetate/d for 17 d. On d 18, three rats from each group were killed to determine baseline (13)C levels. Serum was prepared, and livers were collected and stored at -70 degrees C until analyzed with HPLC and gas chromatography/combustion/isotope ratio mass spectrometry. The remaining rats were supplemented with 52 nmol of (13)C(4)-retinyl acetate. Rats were killed on d 1, 2, 4 and 10. The calculated and measured values of total body reserves (TBR) of vitamin A were within 7% of each other overall, and the relationship was linear (r = 0.98, P < 0.0001). The calculated mean TBR were 0.49 +/- 0.03, 0.82 +/- 0.007 and 3.72 +/- 0.40 micromol, and the measured mean TBR were 0.50 +/- 0.045, 0.69 +/- 0.10 and 3.6 +/- 0.29 micromol for groups L, M and H, respectively. In contrast, serum retinol concentrations did not show a difference among the dietary groups: 1.32 +/- 0.14, 1.35 +/- 0.17 and 1.28 +/- 0.15 micromol/L for groups L, M and H, respectively (P = 0.25). In conclusion, this method offers more sensitivity than traditional methods and may be applicable to human vitamin A status assessment when TBR estimations are desired. PMID- 11053531 TI - Analysis of factors influencing the comparison of homocysteine values between the Third National Health and Nutrition Examination Survey (NHANES) and NHANES 1999+. AB - Two important changes occurred in the time between the Third National Health and Nutrition Examination Survey (NHANES III) (1991-1994) and the later survey (NHANES 1999+) regarding total homocysteine (tHcy), i.e., a change in matrix from serum to plasma and a change in analytical methods. The goals of this study were to determine the magnitude of potential differences between plasma and serum with regard to tHcy concentrations, and between the two analytical methods used in these surveys. Optimally prepared plasma, serum allowed to clot for 30 and 60 min at room temperature and serum allowed to clot for 30 and 60 min and subjected to four freeze-thaw cycles, prepared from blood samples collected from 30 healthy people, were analyzed by both methods. Serum samples had significantly higher tHcy concentrations than plasma samples, and the difference increased with longer clotting time. Freeze-thaw cycles had little or no effect on the variability or bias in the serum sample results. The tHcy results produced by the two analytical methods were significantly different, but consistent across sample types. On average, the results of the method used in NHANES III were lower by 0.64 micromol/L; however, the relative bias varied with tHcy concentration. The tHcy results determined in surplus serum from NHANES III overestimated tHcy concentrations by approximately 10% compared with optimally prepared plasma. The average method bias was 6% between the two analytical methods. On the basis of changes in matrix and methodology, direct comparison of tHcy results between the two surveys is inappropriate. PMID- 11053532 TI - Low arachidonic acid rather than alpha-tocopherol is responsible for the delayed postnatal development in offspring of rats fed fish oil instead of olive oil during pregnancy and lactation. AB - This study was designed to compare in rats the effects of dietary fish oil and olive oil during pregnancy and lactation on offspring development, fatty acid profile and vitamin E concentration. From d 0 of pregnancy, female Sprague-Dawley rats were divided into two groups that were fed purified diets that differed only in their nonvitamin lipid components. One diet contained 10 g fish oil/100 g diet (FOD), whereas the other contained 10 g olive oil/100 g diet (OOD). At d 20 of gestation, maternal adipose tissue fatty acid profile did not differ between rats fed the two diets, whereas both maternal and fetal plasma and liver arachidonic acid (AA) contents were proportionally lower and eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid contents were higher in the FOD group than in the OOD group. alpha-Tocopherol concentration was lower in maternal and fetal plasma, liver and brain in the FOD group than in the OOD group. The postnatal increase in body weight and length was less and body and psychomotor maturation indices were delayed in pups from FOD-fed dams compared with those from OOD-fed dams. This difference was maintained when pups were cross-fostered at birth, with the delay in postnatal development present in the pups suckling dams fed FOD during lactation. At age 21 d, pups suckling dams fed FOD had lower AA and higher EPA and DHA concentrations in brain phospholipids. Although alpha-tocopherol in plasma and liver was lower in pups suckling dams fed FOD rather than OOD, brain alpha-tocopherol concentrations did not differ. Milk yield and milk alpha tocopherol and AA concentrations were lower and EPA and DHA were higher in the milk of dams fed FOD compared with those fed OOD. Postnatal development indices and the proportion of plasma, liver and brain AA concentrations, although not plasma, liver and brain alpha-tocopherol concentrations, recovered to the values found in dams fed OOD when the FOD was supplemented with gamma-linolenic acid. However, postnatal development indices were not recovered when the FOD was supplemented with sufficient exogenous vitamin E to increase plasma and liver alpha-tocopherol concentrations above those in dams fed OOD. Thus, although feeding FOD during pregnancy and lactation decreases both alpha-tocopherol and AA concentrations, the latter deficiency rather than the former seems to be responsible for delayed postnatal development of rat pups. PMID- 11053533 TI - Assessing dermal absorption. AB - The article highlighted in this issue is "Comparative in Vitro-in Vivo Percutaneous Absorption of the Pesticide Propoxur" by Johannes J. M. van de Sandt, Wim J. A. Meuling, Graham R. Elliott, Nicole H. P. Cnubben, and Betty C. Hakkert (pp 15-22). PMID- 11053534 TI - Cytochrome P450, the oxygen-activating enzyme in xenobiotic metabolism. PMID- 11053535 TI - Latex allergy in the workplace. AB - While less than 1% of the general population is sensitized to latex, the U.S. Occupational Safety and Health Administration estimates that 8-12% of health-care workers are sensitized. The major source of workplace exposure is powdered natural rubber latex (NRL) gloves. NRL is harvested from HEVEA: brasiliensis trees and ammoniated to prevent coagulation resulting in the hydrolysis of the latex proteins. Prior to use in manufacturing, the latex is formulated by the addition of multiple chemicals. Thus, human exposure is to a mixture of residual chemicals and hydrolyzed latex peptides. Clinical manifestations include irritant contact dermatitis, allergic contact dermatitis (type IV), and type I immediate hypersensitivity response. Type I (IgE-mediated) NRL allergy includes contact urticaria, systemic urticaria, angioedema, rhinitis, conjunctivitis, bronchospasm, and anaphylaxis. Taking an accurate history, including questions on atopic status, food allergy, and possible reactions to latex devices makes diagnosis of type-I latex allergy possible. To confirm a diagnosis, either in vivo skin prick testing (SPT) or in vitro assays for latex-specific IgE are performed. While the SPT is regarded as a primary confirmatory test for IgE mediated disease, the absence of a U.S. Food and Drug Administration-licensed HEVEA: brasiliensis latex extract has restricted its use in diagnosis. Serological tests have, therefore, become critically important as alternative diagnostic tests. Three manufacturers currently have FDA clearance for in vitro tests, to detect NRL-specific IgE. The commercially available assays may disagree on the antibody status of an individual serum, which may be due to the assay's detecting anti-NRL IgEs to different allergenic NRL proteins. Sensitized individuals produce specific IgE antibody to at least 10 potent HEVEA: allergens, Hev b 1-Hev b 10, each of which differs in its structure, size, and net charge. The relative content and ratios of Hevs in the final allergen preparation most probably could effect diagnostic accuracy. The Hev proteins have been cloned and expressed as recombinant proteins. Sequencing demonstrates both unique epitopes and sequences commonly found in other plant proteins. Sequence homology helps to explain the cross reactivity to a variety of foods experienced by latex allergic individuals. The development of recombinant allergens provides reagents that should improve the diagnostic accuracy of tests for latex allergy. Although clinical and exposure data have been gathered on the factors affecting response in latex-allergic individuals, less is known regarding the development of sensitization. Coupled with in vitro dermal penetration studies, murine models have been established to investigate the route of exposure in the development of latex sensitization. Time-course and dose-response studies have shown subcutaneous, intratracheal, or topical administrations of non-ammoniated latex proteins to induce IgE production. Both in vitro penetration and in vivo studies highlight the importance of skin condition in the development of latex allergy, with enhanced penetration and earlier onset of IgE production seen with experimentally abraded skin. The diagnosis of latex allergy is complicated by these variables, which in turn hinder the development of intervention strategies. Further epidemiological assessment is needed to more explicitly define the scope, trends, and demographics of latex allergy. Diagnostic accuracy can be improved through greater knowledge of proteins involved in the development of latex allergy, and better documentation of the presently available diagnostic tests. In vivo and in vitro models can elucidate mechanisms of sensitization and provide an understanding of the role of the exposure route in latex allergy-associated diseases. Together, these efforts can lead to intervention strategies for reducing latex allergy in the workplace. PMID- 11053536 TI - Comparative in vitro-in vivo percutaneous absorption of the pesticide propoxur. AB - In vitro and in vivo skin absorption of the pesticide propoxur (2 isopropoxyphenyl N-methyl carbamate, commercially Baygon(TM) and Unden (TM); log Po/w 1.56, MW 209.2) was investigated. In vivo studies were performed in rats and human volunteers, applying the test compound to the dorsal skin and the volar aspect of the forearm, respectively. In vitro experiments were carried out in static diffusion cells using viable full-thickness skin membranes (rat and human), non-viable epidermal membranes (rat and human) and a perfused-pig-ear model. Percutaneous penetration of propoxur in human volunteers was measured by analysis of its metabolite (2-isopropoxyphenol) in blood and urine; in all other studies radiolabeled propoxur ([ring-U-(14)C]propoxur) was used. In order to allow for direct comparison, experimental conditions were standardized with respect to dose (150 microg propoxur per cm(2)), vehicle (60% aqueous ethanol) and exposure time (4 h). In human volunteers, it was found that approximately 6% of the applied dose was excreted via the urine after 24 h, while the potential absorbed dose (amount applied minus amount washed off) was 23 microg/cm(2). In rats these values were 21% and 88 microg/cm(2), respectively. Data obtained in vitro were almost always higher than those obtained in human volunteers. The most accurate in vitro prediction of the human in vivo percutaneous absorption of propoxur was obtained on the basis of the potential absorbed dose. The absorbed dose and the maximal flux in viable full-thickness skin membranes correlated reasonably well with the human in vivo situation (maximal overestimation by a factor of 3). Epidermal membranes overestimated the human in vivo data up to a factor of 8, but the species-differences observed in vivo were reflected correctly in this model. The data generated in the perfused-pig-ear model were generally intermediate between viable skin membranes and epidermal membranes. PMID- 11053537 TI - In vitro kinetics of coumarin 3,4-epoxidation: application to species differences in toxicity and carcinogenicity. AB - Coumarin, a natural product and fragrance ingredient, is a well recognized rat liver toxicant, and dietary administration at toxic dosages increased the incidence of rat cholangiocarcinomas and parenchymal liver-cell tumors in a chronic bioassay. Hepatotoxicity in rats is site- and species-specific, and is thought to result from the formation of coumarin 3,4-epoxide and its rearrangement product, o-hydroxyphenylacetaldehyde (o-HPA). The goals of the current study were to describe the in vitro kinetics of the metabolic activation of coumarin, and determine whether species differences in susceptibility to liver injury correlate with coumarin bioactivation determined in vitro. Coumarin 3,4 epoxidation was quantified via the formation of o-HPA in pooled hepatic microsomes from female B6C3F1 mice, male F344 rats, and individual humans (n = 12 subjects), and the apparent kinetic constants for o-HPA production were calculated using nonlinear regression and fitting to either a one-enzyme or two enzyme model. Eadie-Hofstee analyses indicated that o-HPA formation was biphasic in both rat and mouse liver. Although the apparent high affinity K:(m) in rat and mouse liver microsomes was 38.9 and 47.2 microM, respectively, the overall rate of o-HPA formation was far greater in mouse than in rat liver microsomes. Furthermore, the total clearance (CL(int)) of coumarin via o-HPA formation in mouse liver microsomes was 4-fold greater than in rat liver microsomes. Since mice are relatively resistant to hepatotoxicity, the data indicated that rates of o-HPA formation in rat and mouse liver microsomes were not directly predictive of liver toxicity in vivo, and further suggested that o-HPA detoxification played a role in modulating coumarin-mediated toxicity. The current studies also indicated that coumarin 3,4-epoxidation in human hepatic microsomes was minimal. In human liver microsomes (n = 12), the kinetics of o-HPA formation were best described by a single enzyme model, with the K(m) for o-HPA formation ranging from 1320-7420 microM. In the most active human sample, the intrinsic clearance of coumarin via the 3,4-epoxidation pathway was 1/9 and 1/38 that of the rat and mouse, respectively. The in vitro kinetics of o-HPA formation, and in particular, the large quantities of coumarin required for o-HPA production in human liver microsomes, strongly suggest that humans are unlikely to produce toxicologically relevant concentrations of this metabolite following low level coumarin exposures. PMID- 11053538 TI - Reevaluating cancer risk estimates for short-term exposure scenarios. AB - Estimates of cancer risk from short-term exposure to carcinogens generally rely on cancer potency values derived from chronic, lifetime-exposure studies and assume that exposures of limited duration are associated with a proportional reduction in cancer risk. The validity of this approach was tested empirically using data from both chronic lifetime and stop-exposure studies of carcinogens conducted by the National Toxicology Program. Eleven compounds were identified as having data sufficient for comparison of relative cancer potencies from short term versus lifetime exposure. The data were modeled using the chronic data alone, and also using the chronic and the stop-exposure data combined, where stop exposure doses were adjusted to average lifetime exposure. Maximum likelihood estimates of the dose corresponding to a 1% added cancer risk (ED(01)) were calculated along with their associated 95% upper and lower confidence bounds. Statistical methods were used to evaluate the degree to which adjusted stop exposures produced risks equal to those estimated from the chronic exposures. For most chemical/cancer endpoint combinations, inclusion of stop-exposure data reduced the ED(01), indicating that the chemical had greater apparent potency under stop-exposure conditions. For most chemicals and endpoints, consistency in potency between continuous and stop-exposure studies was achieved when the stop exposure doses were averaged over periods of less than a lifetime-in some cases as short as the exposure duration itself. While the typical linear adjustments for less-than-lifetime exposure in cancer risk assessment can theoretically result in under- or overestimation of risks, empirical observations in this analysis suggest that an underestimation of cancer risk from short-term exposures is more likely. PMID- 11053539 TI - Modifying effects of ethinylestradiol but not methoxychlor on N-ethyl-N nitrosourea-induced uterine carcinogenesis in heterozygous p53-deficient CBA mice. AB - It is unknown whether endocrine-disrupting chemicals (EDCs) with estrogenic activities have any modifying effects on uterine carcinogenesis. In our previous study, we established a uterine-carcinogenesis model that is useful for detecting tumor-modifying effects of EDCs by the administration of N-ethyl-N-nitrosourea (ENU) to female heterozygous p53-deficient CBA mice [p53 (+/-) mice]. To investigate the effects of ethinylestradiol (EE) and methoxychlor (MXC) on development of ENU-induced uterine tumors, female p53 (+/-) mice and their wild type littermates [p53 (+/+) mice] received an intraperitoneal injection of 120 mg/kg body weight (bw) of ENU, followed, in Group 1, by no further treatment; in Group 2, by a diet containing 1 ppm EE; in Group 3, by a diet containing 5 ppm EE for 4 weeks and 2.5 ppm EE thereafter; and in Group 4, by a diet containing 2000 ppm MXC for 26 weeks. Uterine proliferative lesions that were induced were composed of both endometrial-stromal and epithelial-cell types. Endometrial stromal sarcomas were induced in p53 (+/-) mice of Groups 1 to 4, and the incidence (87%) in Group 3 was significantly increased compared to Group 1 (47%). Atypical hyperplasias (clear-cell type) of the endometrial gland in p53 (+/-) mice were seen at incidences of 0, 14, 60, and 0% in Groups 1, 2, 3, and 4, respectively, while their incidence in p53 (+/+) mice was 0, 7, 53, and 0%, respectively, with a significant difference between Groups 1 and 3 in both cases. One p53 (+/-) mouse in Group 3 also had an adenocarcinoma consisting of clear cells, and the PCNA labeling indices of the clear-cell atypical hyperplasias, and this endometrial adenocarcinoma, were higher than those of glandular hyperplasias. The present study suggests that 2.5 ppm EE, but not MXC, exerts tumor-promoting effects on stromal and epithelial proliferative lesions of the uteri in p53 (+/-) mice initiated with ENU. PMID- 11053540 TI - The effect of atrazine on puberty in male wistar rats: an evaluation in the protocol for the assessment of pubertal development and thyroid function. AB - Since atrazine (ATR), a chlorotriazine herbicide, has been shown previously to alter the secretion of luteinizing hormone (LH) and prolactin (PRL) through a direct effect on the central nervous system (CNS), we hypothesized that exposure to ATR in the EDSTAC male pubertal protocol (juvenile to peripubertal) would alter the development of the male rat reproductive system. We dosed intact male Wistar rats from postnatal day (PND) 23 to 53 and examined several reproductive endpoints. ATR (0, 12.5, 25, 50, 100, 150, or 200 mg/kg) was administered by gavage and an additional pair-fed group was added to compare the effects of any decreased food consumption in the high dose group. Preputial separation (PPS) was significantly delayed in the 12.5, 50, 100, 150, and 200 mg/kg ATR dose groups. PPS was also delayed in the pair-fed group, although significantly less than in the high dose-ATR group. The males were killed on PND 53 or 54, and pituitary, thyroid, testes, epididymides, seminal vesicles, and ventral and lateral prostates were removed. ATR (50 to 200 mg/kg) treatment resulted in a significant reduction in ventral prostate weights, as did the reduced food consumption of the pair-fed group. Testes weights were unaffected by atrazine treatment. Seminal vesicle and epididymal weights were decreased in the high dose-ATR group and the control pair-fed group. However, the difference in epididymal weights was no longer significantly different when body weight was entered as a covariable. Intratesticular testosterone was significantly decreased in the high dose-ATR group on PND 45, but apparent decreases in serum testosterone were not statistically significantly on PND 53. There was a trend for a decrease in luteinizing hormone (LH) as the dose of ATR increased; however, dose group mean LH was not different from controls. Due to the variability of serum prolactin concentrations on PND 53, no significant difference was identified. Although prolactin is involved in the maintenance of LH receptors prior to puberty, we observed no difference in LH receptor number at PND 45 or 53. Serum estrone and estradiol showed dose-related increases that were significant only in the 200 mg/kg-ATR group. No differences were observed in thyroid stimulating hormone (TSH) and thyroxine (T4) between the ATR groups and the control; however triiodothyronine (T3) was elevated in the high dose-ATR group. No differences in hormone levels were observed in the pair-fed animals. These results indicate that ATR delays puberty in the male rat and its mode of action appears to be altering the secretion of steroids and having subsequent effects on the development of the reproductive tract, which appear to be due to ATR's effects on the CNS. Thus, ATR tested positive in the pubertal male screen that the Endocrine-Disrupter Screening and Testing Advisory Committee (EDSTAC) is considering as an optional screen for endocrine disrupters. PMID- 11053541 TI - A small-volume bioassay for quantification of the esterase inhibiting potency of mixtures of organophosphate and carbamate insecticides in rainwater: development and optimization. AB - The goal of this study was to develop a sensitive in vitro bioassay for quantification of the total esterase inhibiting potency of low concentrations of organophosphate and carbamate insecticides in relatively small rainwater samples. Purified acetylcholinesterase (AChE) from electric eel (Electrophorus electricus) and carboxylesterases from a homogenate of honeybee heads (Apis mellifera) were used as esterases, each having different affinities for the substrates S acetylthiocholine-iodide (ATC) and N-methylindoxylacetate (MIA). MIA hydrolysis by honeybee homogenate was more sensitive to inhibition by organophosphate insecticides than ATC hydrolysis by purified AChE, although the latter parameter is often used for in vitro monitoring of esterase inhibitors. The higher sensitivity of carboxylesterases is attributed to the instant formation of a reversible Michaelis-Menten complex with the inhibitor, which competes with MIA for the active sites of the free enzymes. This dose-dependent instant inhibition can be quantified with kinetics for competitive inhibition at dichlorvos concentrations < 16 nM. At similar concentrations, purified AChE was not instantly inhibited, whereas both AChE and carboxylesterases were irreversibly and progressively inhibited at higher dichlorvos concentrations (IC50(10min) >/= 0.1 microM). Honeybee homogenate mediated MIA hydrolysis was applied as the most sensitive enzyme-substrate combination for experiments with fractionated extracts of 4 rainwater samples collected in a natural conservation area. Most esterase inhibiting potency was found in the polar methanol fraction, with recalculated concentrations equivalent to 12-125 ng dichlorvos per liter rainwater. PMID- 11053542 TI - Comparative metabolism and excretion of benzo(a)pyrene in 2 species of ictalurid catfish. AB - Differential susceptibility of polycyclic aromatic hydrocarbon (PAH)-mediated liver cancer exists in two related species of Ictalurid catfish. Two hypotheses are addressed in this study to explain this difference. Specifically, the relatively insensitive channel catfish 1) do not produce mutagenic PAH metabolites, and/or 2) they more quickly eliminate PAHs due to greater Phase II enzyme activities than the more sensitive brown bullhead. Livers and bile were collected from each species 6, 24, 72, and 168 h after a single 10 mg/kg i.p. benzo(a)pyrene (BaP) exposure. BaP treatment had no significant effect on cytosolic 1-chloro-2,4-dinitrobenzene or ethacrynic acid (EA)-glutathione-S:- transferase (GST) and cis-stilbene oxide-microsomal epoxide hydrolase (EH) activities of either species. Channel catfish EH and GST activities were 1.2-fold higher than brown bullhead activities (p = 0.058 and p < 0.002, respectively). HPLC-APCI-MS of extracted bile and bile enzymatically digested to detect glucuronyl transferase (GT), GST, and sulfotransferase (ST) conjugated metabolites indicated no species differences in elimination or profiles of total biliary metabolites. GT conjugates predominated; ST and GST conjugates were minimal. BaP-diones accounted for the majority of metabolites in both species. Overall, these results indicated that brown bullhead preferentially formed BaP 7,8-dihydrodiol, a precursor to the DNA-reactive BaP-7, 8-dihydrodiol-9,10 epoxide (BPDE), which may be linked to the increased PAH susceptibility in this species. PMID- 11053543 TI - Modeling and predicting selected immunological effects of a chemical stressor (3,4-dichloropropionanilide) using the area under the corticosterone concentration versus time curve. AB - Many chemicals and drugs can induce a neuroendocrine stress response that can be immunosuppressive. Mathematical models have been developed that allow prediction of the immunological impact of such stress responses in mice on the basis of exposure to the important stress-related mediator corticosterone. The area under the corticosterone concentration vs. time curve (AUC) has been used as an indicator of cumulative corticosterone exposure in these modeling studies. In the present study, an immunotoxicant known to induce a stress response, 3,4 dichloropropionanilide (propanil), was evaluated to determine if corticosterone AUC values are related to suppression of immunological parameters in mice treated with this chemical. Linear relationships between corticosterone AUC values and suppression of the following parameters were noted in B6C3F1 female mice: thymus cellularity and thymus subpopulation percentages, splenic subpopulation percentages, natural killer cell activity, MHC class II protein expression, and IgG1 and IgG2a antibody responses to antigen. Linear models derived in previous studies using mice treated with exogenous corticosterone or with restraint stress effectively predicted the immunological effects of 3, 4-dichloropropionanilide on the basis of corticosterone AUC values. The models derived using immobilization stress were more effective (r(2) for observed vs. predicted = 0.90) than the models derived using mice treated with exogenous corticosterone (r(2) for observed vs. predicted = 0.65). This was expected, because most stressors induce a variety of immunomodulatory mediators, not just corticosterone. These findings have implications for risk assessment in immunotoxicology. PMID- 11053544 TI - A single dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin produces a time- and dose dependent alteration in the murine bone marrow B-lymphocyte maturation profile. AB - The halogenated aromatic hydrocarbon, 2,3,7, 8-tetrachlorodibenzo-p-dioxin (TCDD), is a ubiquitous, highly toxic environmental contaminant shown to produce immunotoxic effects in mammals. Although its immunotoxicity has been widely reported, little is known regarding its effect upon the development of immune system cells, especially the B lymphocyte. The present study's purpose was to assess the effect that a single-dose administration of TCDD has, over time, upon bone marrow B-cell progenitors and pro/pre-B-, immature B-, and mature B-cell subpopulations, and to establish a dose-response relationship for these changes. Results showed that the mature B-lymphocyte subpopulation varied in a time dependent manner, with a significant increase one day following TCDD treatment (30 microg/kg body weight [bw]), followed by a significant decrease at day 9 and a return to near-vehicle levels by day 31. Developing and less mature subpopulations were significantly decreased at days 6 and 9. The earliest B cell progenitor subpopulation increased until day 9 and then decreased to vehicle treated levels. Dose response (30, 15, 9, 6, 3, and 0.3 microg TCDD/kg bw) results at 2 days following treatment showed that only the mature-B subpopulation was affected at these doses, and below 6 microg/kg bw no effect was observed. These data suggest that the primary effect of TCDD is on those cells entering, and/or within, the mature B-lymphocyte subpopulation, and the alteration observed in the earlier maturation stages is a compensatory response to the effect on these mature cells. PMID- 11053545 TI - Inhibition of CFU-E/BFU-E by 3'-azido-3'-deoxythymidine, chlorpropamide, and protoporphirin IX zinc (II): a comparison between direct exposure of progenitor cells and long-term exposure of bone marrow cultures. AB - Erythropoiesis occurs in two stages: proliferation amplifies cell number, and differentiation stimulates the acquisition of the functional properties of red blood cells. The erythroid colony-forming unit (CFU-E) amplifies the differentiation process in response to erythropoietic stress in vitro, whereas the burst-forming unit (BFU-E), which is not particularly sensitive to erythropoietin stimulation, gives rise to the CFU-E and, when stimulated, produces morphologically-identifiable erythroid colonies. The aim of this work was to evaluate the toxic effects of the antiviral agent, 3'-azido-3' deoxythymidine (AZT), the antidiabetic drug, chlorpropamide (CLP), and the heme analogous compound, protophorphirin IX zinc (II) (ZnPP), on the proliferation of erythroblastic progenitors by using human umbilical-cord blood cells and murine progenitors from long-term bone marrow cultures. All these agents may interfere with the hemopoietic process, causing myelotoxicity as an adverse effect via different mechanisms. Our results showed selective toxicity of the three drugs on the erythroid progenitors (IC(50): AZT 0.35 +/- 0.13 microM, ZnPP 23.34 +/- 1.16 microM, CLP 1.07 +/- 0.27 mM), with respect to the myeloid progenitors (IC(50): AZT 0.8 microM, ZnPP 103.9 +/- 3.9 microM and CLP > 2800 microM). The IC(50) values were well correlated with peak plasma levels reached in vivo by the drugs. There was a marked similarity between the drug sensitivities of the human and murine progenitors but differences in toxicity exerted by the drugs on the basis of the time of exposure. Drug treatment of long-term cultures, followed by the clonogenic assay of progenitors collected from them in the absence of the drugs, generally resulted in a lower hematotoxicity. PMID- 11053546 TI - Induction of micronuclei following exposure to methylene di-phenyl diisocyanate: potential genotoxic metabolites. AB - Methylene di-phenyl diisocyanate (MDI) is used to make polyurethane products. The predominant occupational disease attributed to diisocyanates, including MDI, is asthma; however, the potential for genotoxicity has also been of concern. Diisocyanates are very reactive compounds that can undergo nonenzymatic hydrolysis to form methylenedianiline (MDA), or react under physiological conditions with primary amines to form ureas and/or with thiols to form labile thiol acid esters. MDA is a carcinogen in animals and a suspected carcinogen in humans. Brown Norway rats (BNR) were exposed to either 7 or 113 mg/m(3) MDI aerosol for 1 h/week x3 weeks and sacrificed 1 week later. Micronuclei (MN) formation was assessed from bone marrow polychromatic erythrocytes (PCE). A dose dependent increase in the frequency of micronucleated polychromatic erythrocytes (MN-PCEs) was noted. In vitro exposure of Chinese hamster lung fibroblasts (V79) to MDA or MDI-thiol conjugates, but not to MDI, significantly increased the frequency of MN. MDI-thiol conjugate-exposed cell cultures did not have detectable levels of MDA. A significant increase in the number of V79 cells in metaphase, as well as the number of cells with precipitants within both the cytoplasm and nuclei, were noted in MDI-glutathione-exposed cultures. The results of this study indicate that MDI aerosol exposure can cause MN formation through either the hydrolysis of MDI to MDA or possibly the formation of thiol conjugates. PMID- 11053547 TI - Protection against Fas receptor-mediated apoptosis in hepatocytes and nonparenchymal cells by a caspase-8 inhibitor in vivo: evidence for a postmitochondrial processing of caspase-8. AB - Lymphocytes can kill target cells including hepatocytes during various inflammatory diseases by Fas receptor-mediated apoptosis. Caspase-8 is activated at the receptor level, thereby initiating the processing of downstream effector caspases. The aim of this study was to investigate the time course of caspase-8 activation and to evaluate the efficacy of the caspase-8 inhibitor IETD-CHO in a model of Fas-induced apoptosis in vivo. C3Heb/FeJ mice were treated with the anti Fas antibody Jo-2 (0.6 mg/kg). Western blot analysis demonstrated increased cytochrome c in the cytosol (20 min), which was followed by the progressive activation of caspase-3, -9 (40-120 min), and caspase-8 (120 min). At 90 and 120 min, extensive hemorrhage was observed, indicating damage to sinusoidal lining cells. In addition, high plasma ALT levels (997 +/- 316 U/L) and histological evaluation indicated severe parenchymal cell injury. Parenchymal and nonparenchymal cells showed a similar increase in caspase-3 activity and DNA fragmentation. Treatment with IETD-CHO (10 mg/kg) attenuated the increase in caspase-3 activity and DNA fragmentation by 80-90% and completely prevented hemorrhage and parenchymal cell damage. IETD-CHO also prevented the early release of mitochondrial cytochrome c and the processing of caspase-3, -8, and -9. Thus, our data support the hypothesis that Fas-mediated apoptosis is dependent on caspase-8 activation in hepatocytes and nonparenchymal cells. However, the bulk of procaspase-8 is processed late, suggesting that only a small amount of procaspase-8 may actually be activated at the Fas receptor. This initial signal may be amplified by further activation of caspase-8 by effector caspases, i.e., after mitochondrial activation. Caspase-8 is a promising therapeutic target for inhibition of Fas-mediated apoptosis. PMID- 11053548 TI - Interactions of the organophosphates paraoxon and methyl paraoxon with mouse brain acetylcholinesterase. AB - The mechanism of acute toxicity of the organophosphorus insecticides has been known for many years to be inhibition of the critical enzyme acetylcholinesterase (EC 3.1.1.7), with the resulting excess acetylcholine accumulation leading to symptoms of cholinergic excess. The bimolecular inhibition rate constant k(i) has been used for decades to describe the inhibitory capacity of organophosphates toward acetylcholinesterase. In the current study, a new approach based on continuous systems modeling was used to determine the appk(i)s of paraoxon and methyl paraoxon towards mouse brain acetylcholinesterase over a wide range of oxon concentrations. These studies revealed that the bimolecular inhibition rate constants for paraoxon and methyl paraoxon appeared to change as a function of oxon concentrations. For example, the appk(i) found with a paraoxon concentration of 1000 nM was 0.16 nM-1h-1, whereas that for 0.1 nM paraoxon was 1.60 nM-1h-1, indicating that the efficiency of phosphorylation appeared to decrease as the paraoxon concentration increased. These data suggested that the current understanding of how these organophosphates interact with acetylcholinesterase is incomplete. Modeling studies using several different kinetic schemes, as well as studies using recombinant monomeric mouse brain acetylcholinesterase, suggested the existence of a second binding site in addition to the active site of the enzyme, to which paraoxon and methyl paraoxon bound, probably in a reversibly manner. Occupation of this site likely rendered more difficult the subsequent phosphorylation of the active site by other oxon molecules, probably by steric hindrance or allosteric modification of the active site. It cannot be ascertained from the current study whether the putative second binding site is identical to or shares common elements with the well-characterized propidium-specific peripheral binding site of acetylcholinesterase. PMID- 11053549 TI - Various nitric oxide donors protect chick embryonic neurons from cyanide-induced apoptosis. AB - The discovery of numerous biochemical effects of cyanide not directly related to the inhibition of the respiratory chain, including the involvement of apoptosis, has challenged the basis of traditional antidote treatment, which primarily depends on nitrite-induced conversion of hemoglobin into methemoglobin, releasing the blockade of cytochrome c oxidase by high-affinity binding of cyanide as cyanmethemoglobin. The fact that amyl nitrite has antidotal effects not related to methemoglobin formation has unfolded new mechanism of actions of nitrites including release of nitric oxide (NO). In this study, we characterized the effect of various NO donor compounds on cyanide-induced cell death in cultured chick embryonic neurons. Apoptosis was induced by treating the neuronal cultures with 1 mM NaCN for 1 h, followed by a cyanide-free incubation period of 23 h. Using this treatment protocol, we showed that cyanide-induced apoptosis was blocked in the presence of the different NO donors sodium nitroprusside, S nitrosoglutathione, S-nitroso-N-acetylpenicillamin, nitroglycerin, 3 morpholinosydnonimine, and diethylamine nitric oxide, indicating independence of the redox-related species of NO released. The effect was confirmed to be mediated by NO, since exhausted NO donors did not afford protection, and the mechanism likely involved chemical modification of thiol groups, since the effect was completely reversed by dithiothreitol. Furthermore, NMDA antagonists protected against cyanide-induced cell death, whereas inhibitors of nitric oxide synthase increased cyanide-induced apoptotic damage, indicating a protective effect of endogenously generated NO, at least in cell cultures. PMID- 11053550 TI - Effect of atrazine on implantation and early pregnancy in 4 strains of rats. AB - Atrazine (ATR) is an herbicide that has been shown to have adverse reproductive effects including alterations in levels of pituitary hormones such as prolactin (prl) and luteinizing hormone (LH) in female LE rats when administered at doses of 200 mg/kg/day for 1 and 3 days. Because the action of prl in promotion of progesterone secretion is essential for the initiation of pregnancy in rats, this study was designed to examine the effect of exposure to ATR during early pregnancy on implantation and short-term pregnancy maintenance. Rats were divided into two groups representing periods of dosing with ATR prior to the diurnal or nocturnal surges of prl. Within each group, four groups consisting of four strains of rats [Holtzman (HLZ); Sprague Dawley (SD); Long Evans (LE); Fischer 344 (F344)] were each further subdivided into four ATR dosages. Rats were dosed by gavage with 0, 50, 100, or 200 mg/kg/day ATR on days 1-8 of pregnancy (day 0 = sperm +). All animals were necropsied on day 8 or 9 of pregnancy. The 200 mg/kg dose of ATR reduced body weight gain in all but one group. Two groups of animals dosed at 100 and 200 mg/kg/day in the nocturnal dosing period showed an increase in percent preimplantation loss, and both of these were F344 rats. HLZ rats were the only strain to show a significant level of postimplantation loss and a decrease in serum progesterone at 200 mg/kg/day both following diurnal and nocturnal dosing. Doses of 100 mg/kg/day also produced postimplantation loss following diurnal and nocturnal dosing, but progesterone levels were decreased only after nocturnal dosing. Alterations in serum LH were seen in several groups. Serum estradiol was significantly increased only in SD rats dosed at the diurnal interval with 200 mg/kg ATR. We conclude that F344 rats are most susceptible to preimplantation effects of ATR and that HLZ rats appear most sensitive to the postimplantation effects of the chemical. LE and SD rats were least sensitive to effects of ATR during very early pregnancy. PMID- 11053551 TI - Analysis of the combined osteolathyritic effects of beta-aminopropionitrile and diethyldithiocarbamate on xenopus development. AB - In order to examine the mechanistic basis between combined effects and mechanisms of action, two osteolathyrogens, beta-aminopropionitrile (betaAPN) and diethyldithiocarbamate (DTC), were tested together on Xenopus embryos. In a separate test, DTC was also tested with copper sulfate to determine the importance of copper in DTC-induced osteolathyrism. Frog embryos (Xenopus laevis) were exposed for 96 h, with daily solution removal and replacement. Preserved tadpoles were evaluated for osteolathyritic lesions. For the betaAPN:DTC test, a 1.2-factor matrix design was used, producing two single chemical and seven mixture-response curves. The chi(2) goodness-of-fit test was used to compare the experimental mixture-response curves with theoretical effects for two combined effects models, dose-addition and independence. All seven mixture curves were consistent with expected results for dose-addition, but the correlations were generally not high. For the DTC:copper test, the three mixture-response curves generated showed that added copper increased the DTC-alone EC(50), but there was no corresponding right shift at the top of the response curves, as observed previously with betaAPN and copper. In the betaAPN:DTC and DTC:copper tests, DTC alone showed a biphasic concentration-osteolathyrism curve, and the slope of the response curve for DTC alone in each test was statistically different than the slope for the betaAPN alone response curve. Taken together, the results suggest the potential for a second osteolathyritic effect of DTC that affected the combined toxicity enough to produce a dose-addition correlation without the chemicals necessarily having the same mechanism. PMID- 11053552 TI - Efficient protection of human bronchial epithelial cells against sulfur and nitrogen mustard cytotoxicity using drug combinations. AB - The aim of this study was to test the efficacy of several candidate molecules against sulfur mustard (SM) and nitrogen mustard (HN2) using a human bronchial epithelial cell line (16HBE14o-). Candidate molecules were chosen on the basis of the known cytotoxicity mechanisms of mustards or their efficacy previously observed on other cellular models. It included the sulfhydryl-containing molecules N-acetyl-cysteine (NAC) and WR-1065, the nucleophile hexamethylenetetramine (HMT), the energy-level stabilizer niacinamide (NC), the antioxidant dimethylthiourea (DMTU), L-arginine analogues such as L thiocitrulline (L-TC) and L-nitroarginine methyl ester (L-NAME), and the anti gelatinase doxycycline (DOX). Their efficacy was determined using 2-(4-[3 iodophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2Htetrazolium (WST-1) reduction by viable cells 24 h after initial exposure to 100 microM HN2 or SM. On individual immediate cotreatment, some molecules exhibited selective protection against only one mustard, such as DMTU and WR-1065 against HN2 and DOX against SM, whereas NAC and L-TC were effective against both SM and HN2 cytotoxicity. However, as the level of protection against SM was always weak compared to HN2, several combinations were investigated against SM to improve the protection. The effective combinations (L-TC + DOX, NAC + DOX, NAC + DMTU, NAC + HMT, NC + DOX) combined agents, reducing the bioavailability of the mustard with compounds possibly acting on the consequences of alkylation. One of these combinations, NAC + DOX, appeared to be the most interesting, as these agents are already used in human therapy. It exhibited good efficacy in delayed cotreatment (up to 90 min) against SM. PMID- 11053553 TI - Disposition of [Ring-U-(14)C]styrene in rats and mice exposed by recirculating nose-only inhalation. AB - The disposition of styrene was studied in a group of 12 Sprague Dawley rats and two groups of 30 CD1 mice exposed separately to 160 ppm [ring-U-(14)C]styrene of high specific radioactivity of 1.92 TBq x mol(-1) (52 Ci x mol(-1)) for 6 h. A nose-only exposure system was successfully adapted to (1) recirculate a portion of the flow to limit the amount of (14)C-styrene required, and (2) avoid any polymerization of the compound. The mean uptake of styrene in rats was 113 +/- 7 micromol x kg(-1) x h(-1) and stable over time. The mean uptake in mice was higher, 189 +/- 53 and 183 +/- 76 micromol x kg(-1) x h(-1), for the first and second mouse inhalation experiment, but decreased steadily over time. Some of the mice, but none of the rats, showed signs of overt toxicity. The overall excretion of styrene and its metabolites was quantitatively similar in rats and mice. Urinary excretion was the primary route of excretion while fecal excretion accounted for only a very small part of the radioactivity. There was, however, a significant difference between mice and rats in the exhalation of (14)CO(2), which must have resulted from opening and subsequent breakdown of the aromatic ring. In mice the exhalation of (14)CO(2) accounted for 6.4 +/- 1.0 and 8. 0 +/- 0.5% of the styrene retained during the first and second mouse inhalation experiment. In rats, exhalation of (14)CO(2) accounted for only 2.0 +/- 0.7% of the retained styrene. Together with the results from the quantitative whole-body autoradiography (showing significantly higher binding in mouse lung and nasal passages compared to rat) the larger production of (14)CO(2) might be indicative of the formation of reactive ring-opened metabolites in the mouse lung, which, in turn, might be related to the observed development of bronchioalveolar tumors and nasal effects in mice exposed to styrene. PMID- 11053554 TI - Inhalation toxicity of 1,6-hexamethylene diisocyanate homopolymer (HDI-IC) aerosol: results of single inhalation exposure studies. AB - The early acute pulmonary response of female Wistar rats exposed nose-only to a mixture of 1,6-hexamethylene diisocyanate homopolymer (HDI-IC) aerosol was examined. This study was designed to investigate the time course of the relationship between acute pulmonary irritation and ensuing disturbances of the air/blood barrier in rats exposed to concentrations of 3.9, 15.9, 54.3, or 118. 1 mg HDI-IC/m(3). The duration of exposure was 6 h, followed by serial sacrifices 0 h, 3 h, 1 day, 3 days, and 7 days postexposure. Concentrations were selected based on the results of a 4-h acute inhalation study in rats (LC(50) = 462 mg/m(3)). Bronchoalveolar lavage (BAL) fluid was analyzed for markers indicative of injury of the bronchoalveolar region, including phospholipids as proxy of altered surfactant homeostasis. Glutathione (GSH) was determined in BAL fluid and lung tissue. BAL cells with increased intracellular phospholipids were observed on day 1 and especially day 3, with some residual increase on day 7. Increased intracellular phospholipids and activity of acid phosphatases appear to suggest that phagocytized phospholipids may transiently affect lysosomal function. Following exposure to 15.9 mg/m(3), changes returned almost entirely to the level of the air-exposed control on day 7. Especially at higher exposure concentrations, lung weights and total number of cells in BAL were still statistically significantly elevated at this time point. Experimental evidence suggests that markers indicative of a dysfunction of the air/blood barrier, such as angiotensin-converting enzyme, total protein, and phospholipids engulfed by alveolar macrophages, were most sensitive to probe this type of changes. Although GSH in BALF was increased following exposure, there was an apparent depletion of tissue GSH immediately after cessation of exposure. In summary, this study suggests that respirable HDI-IC aerosol appears to cause a transient dysfunction of the air/blood barrier indicated by an increased extravasation of plasma constituents. Despite the remarkable extent of effects observed, most changes were reversible within a postexposure period as short as 7 days. First evidence of increased leakage of pulmonary epithelial barrier was observed at 3.9 mg/m(3). With respect to changes of early markers of pulmonary epithelial barrier dysfunction, approximately 3 mg HDI-IC/m(3) was considered to be the threshold concentration for acute pulmonary irritation. PMID- 11053555 TI - Upper respiratory tract toxicity of inhaled methylvinyl ketone in F344 rats and B6C3F1 mice. AB - The National Toxicology Program is conducting a chemical class study to investigate the structure-activity relationships for the toxicity of alpha,beta unsaturated ketones. Methylvinyl ketone (MVK) was selected for study because it is a representative straight-chain aliphatic alpha,beta-unsaturated ketone and because of its extensive use and widespread exposure. Short-term inhalation studies of MVK were conducted to provide toxicity data for comparison with other alpha,beta-unsaturated ketones and for use in designing chronic toxicity and carcinogenicity studies. In 2-week studies, rats and mice were exposed to 0, 0.25, 0.5, 1, 2, 4, or 8 ppm MVK 6 h/day, 5 days/week for 12 exposures. Morbidity and early deaths occurred in all male and female rats after 1 exposure and in 2 male mice after 10 exposures to 8 ppm. Rats exhibited nasal cavity toxicity and lung necrosis at 4 ppm. No toxicity was observed in animals exposed to less than 2 ppm. Based on these results a 13-week study was conducted at 0, 0.5, 1, and 2 ppm MVK. As observed in the 2-week study, the nasal cavity was the main target organ and rats were more sensitive than mice. Respiratory and olfactory epithelial necrosis were prominent by day 21 in the rat. At study termination these lesions were still evident but not as severe as noted earlier. Additionally, changes such as olfactory epithelial regeneration and metaplasia (respiratory) as well as respiratory epithelial hyperplasia and metaplasia (squamous) were clearly evident. Nasal lesions in mice were limited to a subtle squamous metaplasia of transitional and/or respiratory epithelium covering predominantly the tips of naso- and maxilloturbinates in Levels I and II. A transient, leukopenia was observed in rats exposed to 2 ppm, however, this effect was not present after 13 weeks of exposure. In mice, leukocyte counts were significantly decreased at all exposure concentrations after 13 weeks of exposure. Absolute testicular and epididymal weights and sperm counts were decreased at the high dose only. MVK can be characterized as a reactive, direct acting gaseous irritant. MVK exposure causes the same nasal cavity lesions as the cyclic alpha,beta-unsaturated ketone, 2-cyclohexen-1-one, although at lower exposure concentrations. PMID- 11053556 TI - The effects of diet, ad Libitum feeding, and moderate and severe dietary restriction on body weight, survival, clinical pathology parameters, and cause of death in control Sprague-Dawley rats. AB - A 2-year study was conducted in Sprague-Dawley rats to compare the effects of ad libitum (AL) feeding and dietary restriction (DR) on body weight, survival, cause of death, and clinical pathology parameters. Three groups of 120 rats/sex each received the following daily rations of a maintenance rodent diet: ad libitum (AL group); 75% of adult AL food consumption (25% DR group); and 45% of adult AL food consumption (55% DR group). Among the 3 groups, there were generally no differences in relative (food intake per gram of body weight) food consumption. Compared to the AL group, decreased body weight gain occurred in DR groups and was associated with an increase in survival proportional to the DR rate. The main cause of death was pituitary adenomas in all groups. Decreases in total leukocyte, segmented neutrophil, lymphocyte, and platelet counts occurred in the 55% DR group. In serum biochemistry, there were decreases in total protein, albumin, total and HDL cholesterol, and total calcium, and increases in alkaline phosphatase activities and chloride in 55% DR females, as well as decreases in triglycerides in the 55% DR group and in 25% DR females. Results of urinalyses showed decreases in urine volume and protein, and increases in urinary pH in both DR groups. In conclusion, a DR rate of approximately 25% appears to be appropriate for Sprague-Dawley rats in toxicity and carcinogenicity assays to improve survival without impairing growth and routine clinical pathology parameters. PMID- 11053557 TI - Lipopolysaccharide augments aflatoxin B(1)-induced liver injury through neutrophil-dependent and -independent mechanisms. AB - Exposure to small, noninjurious doses of the inflammagen, bacterial endotoxin (lipopolysaccharide, LPS) augments the toxicity of certain hepatotoxicants including aflatoxin B(1) (AFB(1)). Mediators of inflammation, in particular neutrophils (PMNs), are responsible for tissue injury in a variety of animal models. This study was conducted to examine the role of PMNs in the pathogenesis of hepatic injury after AFB(1)/LPS cotreatment. Male, Sprague-Dawley rats (250 350 g) were treated with either 1 mg AFB(1)/kg, ip or its vehicle (0.5% DMSO/saline), and 4 h later with either E. coli LPS (7. 4 x 10(6) EU/kg, iv) or its saline vehicle. Over a course of 6 to 96 h after AFB(1) administration, rats were killed and livers were stained immunohistochemically for PMNs. LPS resulted in an increase in PMN accumulation in the liver that preceded the onset of liver injury. To assess if PMNs contributed to the pathogenesis, an anti-PMN antibody was administered to reduce PMN numbers in blood and liver, and injury was evaluated. Hepatic parenchymal cell injury was evaluated as increased alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities in serum and from histologic examination of liver sections. Biliary tract alterations were evaluated as increased concentration of serum bile acids and activities of gamma glutamyltransferase (GGT), alkaline phosphatase (ALP), and 5'-nucleotidase (5' ND) in serum. Neutrophil depletion protected against hepatic parenchymal cell injury caused by AFB(1)/LPS cotreatment but not against markers of biliary tract injury. This suggests that LPS augments AFB(1) hepatotoxicity through two mechanisms: one of which is PMN-dependent, and another that is not. PMID- 11053581 TI - First hemodialysis access selection varies with patient acuity. AB - Timely placement of a reliable permanent vascular access is essential for hemodialysis care quality; National Kidney Foundation Dialysis Outcomes Quality Improvement (NKF-DOQI) guidelines emphasize native arterio-venous (AV) fistulae as preferred access for incident patients. As part of Network One's Vascular Access Quality Improvement Project (QIP) we investigated whether patients' course to end-stage renal disease (ESRD) influenced vascular access selection. Baseline information was obtained for incident (1998) dialysis patients from 6 centers participating in the Network QIP. Patients were subdivided into 3 predefined clinical groups: KNOWN (known chronic renal disease, seen by a nephrologist, with predictable progression to ESRD), CRISIS (KNOWN but with unanticipated medical crisis precipitating ESRD), and UNKNOWN (not known to have chronic renal insufficiency or never seen by a nephrologist before developing ESRD). Two hundred forty patients were identified (median age 69.9, 42% diabetic). Only 43% of the entire population experienced an orderly progression to renal insufficiency. The most frequent initial access was a catheter (54%), followed by a fistula (29%) and a graft (16%), but selection of initial access differed significantly by patient group, with 46% of KNOWN patients receiving a fistula (P <.001). After 2 months of dialysis, the initial access supported dialysis in only 53.7% of the KNOWN patients, and in 59.4% and 45.7% of the CRISIS and UNKNOWN patients, respectively. We conclude that unpredicted, new ESRD patients are common and are less likely to receive a fistula as initial hemodialysis access. Studies should define optimum access management when dialysis requirement is unforeseen. PMID- 11053582 TI - Barriers to the delivery of adequate hemodialysis in ESRD Network 4. AB - Dialysis dose has been established as a determinant of morbidity and mortality in chronic hemodialysis patients. To identify remediable barriers to the delivery of adequate hemodialysis, we examined factors that affected adherence to prescribed dialysis dose. End-Stage Renal Disease (ESRD) Network 4 facilities that fell into the lowest quintile in measures of dialysis adequacy were studied. At the time of this study, Network 4 was composed of 178 dialysis facilities in Delaware and Pennsylvania. Those 29 facilities had an average delivered urea reduction ratio (URR) of <0.67 and/or 71% of patients with a URR of 0.65. (The mean URR value of Network 4 was 0. 699 with a compliance ratio of 80%.) Dialysis treatment sheets were reviewed for all patients in the 29 facilities for all treatments during a calendar week. Predialysis and postdialysis blood urea nitrogen (BUN) values from 1 treatment during this week were used to calculate URR and Kt/V. A total of 1,339 patients with a mean age of 61.9 +/- 15.1 years and a mean duration of ESRD of 3.4 +/- 3.3 years were dialyzed in the 29 units. Mean prescribed duration of dialysis (T) was 219 +/- 26 min. with a mean blood flow rate (BFR) of 393 +/- 62 mL/min. Concordance between the prescribed and delivered T (-5 min), BFR (-50 mL/min), and hemodialyzer were assessed, by patient, for each treatment (Tx). Characteristics of a delivered Kt/V < 1.2 were duration <4 hours, BFR < 350 mL/min, patient weight > 100 kg, and delivered BFR 50 mL/min less than prescribed BFR. Multivariate analysis of the relationship between delivered dose of dialysis and patients and treatment characteristics identified black race, male gender, and younger age as demographic factors associated with low delivered dose. Potential remediable barriers identified by this analysis included reduced treatment time (>10%) and use of catheters for angioaccess. These data suggest components of the dialysis process that might be targeted in future quality improvement projects to improve the adequacy of dialysis delivery. PMID- 11053583 TI - Improving compliance with the dialysis prescription as a strategy to increase the delivered dose of hemodialysis: an ESRD Network 4 quality improvement project. AB - Delivery of an inadequate dose of hemodialysis is associated with a significant increase in the relative risk of both hospitalization and death. We hypothesized that noncompliance with the dialysis prescription, defined as failure to achieve the prescribed blood flow, failure to dialyze for the prescribed duration, or failure to use the prescribed dialyzer, was a significant factor in patients not achieving a urea reduction ratio (URR) of > or =0.65. We identified the 29 dialysis facilities in ESRD Network 4 that had the lowest average URR and/or lowest percent of patients with a URR > or =0.65 based on quarterly data reports. Each facility was surveyed by review of all dialysis treatment sheets from a single week by network staff to evaluate for noncompliance with the dialysis prescription. Facility-specific data were reported back to each facility. Each facility was required to develop a facility-specific quality improvement plan after receiving intensive education on the quality improvement process. After 9 months the facilities were resurveyed. Although the compliance with the dialysis prescription decreased from 54.0% to 53.6% (P =.026), the delivered URR increased from 0.679 +/- 0.072 to 0.688 +/- 0.070 (P =.026) with an increase in the percentage of patients with a URR > or = 0.65 from 69.7% to 75% (P =.0096). Kt/V increased from 1.37 +/- 0.26 to 1.41 +/- 0.27 (P =. 0001). Analysis of the process changes instituted by the individual facilities showed an increase in the prescribed dose of dialysis. Thus, although the process goal of improved compliance with the dialysis prescription was not achieved, the outcome goal of an increased delivered dose of dialysis was met through an alternative process change of an augmented dialysis prescription. PMID- 11053584 TI - 1999 facility Profile Reports. AB - In November 1999, the Mid-Atlantic Renal Coalitions (MARC) developed and distributed its first version of a facility-specific Profile Report to the medical directors and head nurses of dialysis facilities in Washington, DC, Maryland, Virginia, and West Virginia. The profiles were distributed to support the facilities' internal quality improvement efforts and were not intended to be released as public documents. Facility characteristic and profiling indicators were selected based on the availability, reliability, and value of the data. The Profile Report consisted of 2 data displays with corresponding definition tables. An evaluation mechanism was discussed, but it was determined that the initial report would not be formally evaluated and that this would be included as a feature of future versions. Future versions will be expanded, as additional data become available, and will include rankings and data trending where appropriate. PMID- 11053585 TI - Adequacy of peritoneal dialysis: a quality improvement project of The Renal Network, Inc (Illinois, Indiana, Kentucky, and Ohio). AB - The Renal Network, Inc, (end-stage renal disease [ESRD] Networks 9 and 10) from the states of Illinois, Indiana, Kentucky, and Ohio includes approximately 4,250 peritoneal dialysis patients. To better assess the state of peritoneal dialysis in Networks 9 and 10, we undertook a peritoneal dialysis prescription adequacy quality improvement project. Adequacy and facility practice information were obtained from a 100% sample of peritoneal dialysis patients in the Networks. The mean total Kt/V urea for those facilities with a written policy for adequacy measurement was 2.18 +/- 0.66 versus a mean of 2.13 +/- 0.70 for those facilities without a written policy (P = not significant [NS]). The mean total Kt/V urea for those facilities with a written procedure was 2.17 +/- 0.66 versus a mean of 2.12 +/- 0.70 for those facilities without a written procedure (P = NS). There was a significant positive relationship between the observed total Kt/V urea and the frequency of measurement (P <.001). No relationship existed between the value of the D/P creatinine ratio and the weekly dialysis Kt/V. This study supports the recommendation of frequent measurement of peritoneal dialysis adequacy. PMID- 11053586 TI - Use of transplant status codes to monitor access to kidney transplantation. AB - The kidney transplantation process involves a series of steps related to interest in transplantation, medical suitability, pretransplant workup, and eventual transplantation. We developed transplant status codes to monitor the progress of dialysis patients in Ohio, Kentucky, Indiana, and Illinois as they move through these steps. Of the first 2,137 dialysis patients monitored with these new codes, 29% were interested in receiving a transplant, 31% were medically suitable to receive a kidney transplant, 15% had completed a pretransplant workup, 13% were on a cadaveric transplant waiting list, and 2% were awaiting a living-related transplant. Examination of these steps by gender showed differences in interest (men 32% v women 26%, P =.01) and medical suitability (men 33% v women 28%, P =.01). The proportion of patients successfully completing each step differed across facilities. In conclusion, transplant status codes are a useful method to monitor access to transplantation. As part of a facility-specific quality improvement program, such codes may be used to develop targeted interventions to help patients move through the transplant process as expeditiously as possible. PMID- 11053587 TI - Managing the lifeline: preemptive access management for better outcomes for hemodialysis patients and programs. Medical Review Board of The Renal Network, Inc. AB - Problems with vascular access for hemodialysis patients contribute substantially to the morbidity, mortality, and high costs associated with renal replacement therapy. Data from the Network 9/10 Regional Hemodialysis Vascular Access Quality Improvement Project show evidence of incremental improvements, with more native vein fistulae and fewer grafts. However, increased numbers of catheters were observed, and still fully 32% of the catheters in place were associated with no internal access created after more than 90 days on dialysis. This article reviews recent contributions to understanding patterns of access-related care, pathophysiology of vascular access complications, and new approaches to achieving the preferred native vein arteriovenous fistulae. A financial analysis shows that dialysis units that employ dedicated access management personnel can expect bottom-line benefits that will easily cover the added expense. These benefits will be in addition to improvements in morbidity and mortality for patients. PMID- 11053588 TI - Wheels within wheels: creating a circle of knowledge through communication. AB - For end-stage renal disease patients, patient education is a critical aspect of treatment and wellness. A successful patient education program depends on knowing the target audience and choosing materials and media according to what they already know and what they want and need to learn. Understanding how people learn and selecting an educational model to guide the creation of an educational program provide the underpinnings for an effective presentation. Selection of materials and medium and determining who, when, where, and how the program is conducted optimize the potential for learning. This article presents and illustrates part of The Renal Network's (ESRD Networks 9/10) outreach, communication, and programming efforts to bring all of these elements together in educational activities suitable for a diverse and significantly older population of patients across a 4-state area. It highlights The Renal Network's use and support of the Patient Advisory Council Representative as a patient leader undertaking activities on the facility level to provide patient-to-patient assistance, information sharing, and educational opportunities for fellow patients at their facility. This article shows how the Network initiated a consistent flow of information to facilitate the identification, assessment, selection, media, and means for its patient education activities. PMID- 11053589 TI - Unraveling the realities of vascular access: the Network 11 experience. AB - Arteriovenous (AV) fistulae are well recognized as the preferred vascular access for hemodialysis, yet national data show that only 23% of patients used an AV fistula in 1997. To identify barriers to the placement of native AV fistulae, the Renal Network of the Upper Midwest, Inc. (End-Stage Renal Disease [ESRD] Network 11) initiated a vascular access project to look at the process of referral for patients beginning hemodialysis in the first 6 months of 1999. Of these patients, 63% began hemodialysis with a catheter as the only access, 22% had an AV fistula placed (but only 14% used an AV fistula for their first dialysis treatment), and 15% began with a graft. About 40% of patients were referred to a nephrologist less than 1 month before dialysis, allowing little chance for permanent access placement. Yet 27% of patients used a catheter on the first hemodialysis treatment and were seen by a nephrologist more than 1 month before starting dialysis, indicating the presence of an opportunity to improve. At 6 months after initiation of dialysis, 25% of patients who began dialysis using a catheter were using an AV fistula and 35% were using a graft. Network 11 plans to use this information to promote early referral of patients to a nephrologist and subsequent prompt referral of such patients to a vascular surgeon. Other activities to improve vascular access management are also indicated. PMID- 11053590 TI - Network 12 hepatitis B vaccination quality improvement program: an educational program directed at physicians, staff, and patients. AB - Hepatitis B is easily spread via contact with infected blood. Hemodialysis patients and staff are particularly at risk for acquiring hepatitis B. Consequently, vaccination of hemodialysis patients and staff is strongly recommended. However, the vaccination rate among dialysis patients in this country remains below 50%. End-Stage Renal Disease (ESRD) Network 12 developed a quality improvement project directed at increasing patient vaccination by regular surveys, reports, and education of physicians, staff, and patients. Seventy-seven percent of facilities in the 4-state Network participated. Overall vaccination rate increased from 66.9% to 73.2% over 18 months (P <.05). The greatest improvement was seen among units with less than 60% of patients vaccinated initially, with mean facility vaccination rate increasing from 31.2 +/- 20.5% to 57.5 +/- 30.1% in the last available data period (P <.001). Only 3 of these 29 units failed to improve. The 90 units that had 60% to 97% vaccination rates initially improved significantly from 79.8 +/- 9.6% to 82.4 +/- 15% (P =.015). Three quarters of these units showed improvement. Only units with 100% vaccination deteriorated, but still maintained vaccination rates of 74.5 +/- 25.6%. An education-based quality improvement project can improve the hepatitis B vaccination rate of hemodialysis patients. PMID- 11053591 TI - Mission possible: vascular access-decreasing the use of catheters in the Texas hemodialysis community. AB - In 1999, the End-Stage Renal Disease Network of Texas, Inc, #14 (NW14) initiated a quality improvement project in support of the Health Care Finance Administration's End-Stage Renal Disease Health Care Quality Improvement Program. The project, titled "Mission Possible: Vascular Access: Decreasing the Utilization of Catheters in the Texas Hemodialysis Community," aims to assist the Texas dialysis community decrease the use of hemodialysis catheters. This report reviews the goal of the project, baseline vascular access survey results, and quality improvement activities initiated to help dialysis professionals implement quality management processes that will improve the quality of care for Texas dialysis patients. PMID- 11053592 TI - Influenza immunization rates in the Intermountain End-Stage Renal Disease Network (Network 15). AB - Billing data from the Health Care Financing Administration (HCFA) indicated that the influenza immunization rates for dialysis patients in the United States do not meet the goal of 60% set by Healthy People 2000, and fall significantly short of the goal of 90% of all Medicare beneficiaries as outlined in Healthy People 2010. Influenza and pneumonia together are the sixth leading cause of death in the United States. Despite the known benefits of influenza vaccination in reducing morbidity and mortality, only 40% to 50% of high-risk patients are immunized. Although HCFA/Medicare billing data may not provide the best measurement of actual practice, it is currently the only measure available from any national source. The data suggest that there is a need for improvement. Because the HCFA/Medicare rates were based only on those immunizations for which Medicare was billed, End-Stage Renal Disease Network 15 embarked on a project to determine a more accurate rate of immunization within the Network based on information provided by the dialysis facilities. Influenza vaccination rates for the winter 1998 flu season ranged from 51.5% to 84.9% for the states in the Network; the rate for the whole Network was 74.6%. The HCFA/Medicare billed influenza immunization rates were 26.5 to 45.6 percentage points lower. PMID- 11053593 TI - Improving adequacy of hemodialysis in Northern California ESRD patients: a final project report. Provider Participants and Medical Review Board of the TransPacific Renal Network. AB - The National Core Indicators Project, initiated in 1994, has brought progressive changes in adequacy of dialysis for end-stage renal disease (ESRD) patients in the TransPacific Renal Network and across the United States. The 1998 Core Indicator Project showed each Network's standing for percentage of patients with urea reduction ratio (URR) > or = 0.65 and average URR. The TransPacific Renal Network ranked 12(th) among the 18 Networks for this adequacy measure. The goals of this project were to improve the Network standing in the United States for the percent of patients with URR > or = 0.65, eliminate or reduce the barriers to achieving adequate dialysis, and evaluate URR versus KT/V data and the variances occurring with these measures. In January 1999, data were collected from all 113 Northern California hemodialysis facilities for quarter 4, 1998, to evaluate adequacy. Each facility provided patient population (N) for KT/V and URR samples, facility averages for KT/V and URR, number of patients with KT/V > or = 1.2 and URR > or = 0.65, and data on post-blood-urea-nitrogen (BUN) sampling methods. A random selection of 10% (12) providers with data below the US and Network standards was selected for an intensive assessment. Using baseline measurements, on-site data were collected from a random selection of the patient population. Chart data were reviewed, analyzed, and discussed in an exit interview with the facility management. On-site visits were performed in July/June 1999. The primary focus included adequacy data and process of care that affect adequacy outcomes, concurrent review of patients receiving treatment at the time of the site visit, and general medical record review. In Phase I, only 12 facilities showed an average URR below 0.65. All facilities reported an average KT/V greater than the DOQI target of 1.2. Forty-two facilities had their percentage of patients with a URR below the national benchmark; only 18 facilities had their percentage of patients with a KT/V below the national benchmark. Only 9% (n = 8) of the 113 providers had a variance in post-BUN sampling methodologies that could be related to the clinical measure of adequacy. In Phase II, a random selection of 12 providers with data below US and Network standards was made for an intensive assessment. A total of 217 patient records were reviewed from a population of 1,027. In addition to comparison of baseline data, each facility was assessed for barriers to achieving adequacy outcomes. The number of problems was extensive and specific to each facility; however, a common reoccurring theme in the majority of events was the lack of supporting documentation for changes to the plan of care when variances occur. The most common occurrences were incorrect blood flow and dialysate flow with no supporting documentation on record for the prescription not being met. In Phase III, Network interventions for facilities not meeting US and Network standards for adequacy as measured by URR and KT/V included required quarterly reporting on their facility-specific quality improvement programs for adequacy. In addition the 12 facilities that participated in the intensive assessment had additional interventions that included an educational "tool box" focused on documentation, legal implications of charting, and general medical records management, and an educational program to review information to be shared with facility staff. All on-site facilities reported ongoing quality improvement programs. In some facilities they did provide a focus on processes and not only a measurement of an indicator. All facilities reported a team concept of some type used in their program. Although there were similarities in the facilities, each facility presented with a unique combination of barriers. In addition to a large patient-to-RN ratio, the lack of technical education for the unlicensed assistive personnel on processes and outcomes appears to play a significant role in the achievement of PMID- 11053594 TI - Improved hepatitis B vaccination rates in ESRD patients in California. AB - According to the Centers for Disease Control (CDC) Survey of Dialysis Associated Diseases, California, which includes Network 17 and 18, had one of the lowest hepatitis B vaccination rates in the country for 1994, 1995, and 1996. With 3 outbreaks of hepatitis B (HBV) in California in 1994, hepatitis B vaccination was chosen as a quality improvement project in both Network 17 and 18. With input from both Medical Review Boards and HCFA Region X, a project was formulated which focused on the improvement of the number of facilities which had hepatitis B vaccination rates which are greater than 50%. The overall purpose of both projects was to: (1) achieve access to preventative services for end-stage renal disease (ESRD) Medicare beneficiaries; (2) increase the number of ESRD patients in California who are vaccinated for HBV; (3) eliminate dialysis in California as an independent risk factor for contracting HBV; (4) decrease the number of ESRD facilities with HBV vaccination rates of 0%; and (5) increase the number of ESRD facilities with HBV vaccination rates greater than 50%. In 1998, both Network 17 and 18 denominators were adjusted to reflect the population which is eligible for vaccination. Because of historically low vaccination rate in California, the 1998 data collection sought to ascertain precise numbers for the ESRD patient population. Data were used from the 1996 and 1997 CDC Survey of Dialysis Associated Diseases from baseline measurements of HBV vaccination rates for all facilities in both Network 17 and 18. The CDC did not conduct a survey in 1998, however, Network 17 and 18 conducted a survey of dialysis associated diseases for all of California ESRD facilities. A data collection tool was designed to gather information on processes and outcomes in each facility. This allowed a continuous quality improvement (CQI)-based approach to analyze the problem, where tools like cause/effect and Pareto diagrams provided information on factors and issues affecting low HBV vaccination rates. Interventions were designed to target those specific factors. Interventions included creation of the "Hepatitis Booklet" (Network 18) and the "Hepatitis Resource Guide" (Network 17); mailing of the resource material to all providers (Network 18), and with vaccination rates less than 50% (Network 17); development of facility specific profiles; and policy statements by both Medical Review Boards on Hepatitis B Vaccination. The number of ESRD patients in California who are vaccinated for HBV increased to 53% or 11,412 patients of 21,617 eligible patients in both Networks. The number of ESRD patients in California who are vaccinated plus those in the process of receiving the series brought the California vaccination rate of 72% or 15,653 for 21,617 eligible patients in both Networks. The number of ESRD facilities in California with HBV vaccination rates of 0% decreased to 10 facilities in 1998, from 75 facilities in 1997, and 135 facilities in 1996. The number of ESRD facilities in California with HBV vaccination rates more than 50% increased to 175 facilities, from 87 facilities in 1997, and 52 in 1996. The number of patients developing antibodies post-vaccine was 62% (Network 18). Facilities in Network 17 with vaccination rates exceeding 50% who did not receive the Hepatitis B Resource Guide vaccinated 44% of all patients vaccinated or in progress in Network 17 in 1998. Facilities in Network 17 with vaccination rates less than 50% who did receive the Hepatitis B Resource Guide vaccinated 57% of all patients vaccinated or in progress in Network 17. For the first time, vaccination rates were collected on peritoneal dialysis (PD) patients. In Network 17, 51% of PD patients are vaccinated versus 59% of hemodialysis patients. In Network 18, 48% of PD patients are vaccinated versus 48% of hemodialysis patients. Resource material and feedback reports developed by both Networks facilitated improvements in Hepatitis B vaccination of ESRD patients in Ca PMID- 11053595 TI - Patient safety: the challenges and opportunities for the ESRD program. AB - The Institute of Medicine's report, issued December 1999, entitled "To Err is Human; Building a Safer System," describes the magnitude of the problem of errors in medicine and carts an agenda for improving patient safety. The essential features include establishing patient safety as a national focus, identifying and learning from errors, passing legislation to protect reporting, and adopting the patient safety sciences. The presidential report of February 2000, issued by the Quality Interagency Task Force (QuIC), formulates a federal government response across all federal agencies. The challenges and opportunities facing the end stage renal disease (ESRD) Program and the ESRD Network Organizations include taking a leadership role, raising awareness, conducting educational programs, and facilitating making errors visible, for the purposes of learning and improvement. PMID- 11053596 TI - A pilot study: stability of psychiatric diagnoses over 6 months in burning mouth syndrome. AB - Ten people with burning mouth syndrome (BMS) were interviewed by a psychiatrist using the Schedules of Clinical Assessment in Neuropsychiatry (SCAN) at initial presentation and at 6-month follow-up. A range of psychiatric disorders from the neurotic spectrum was identified using the International Classification of Diseases (ICD-10) criteria, but the diagnoses were unstable. Six of the ten individuals received a psychiatric diagnosis, suggesting that the prevalence of psychiatric morbidity is high in this common dental syndrome. Psychiatric aspects of BMS require further investigation. PMID- 11053597 TI - The timing of acts of deliberate self-harm: is there any relation with suicidal intent, mental disorder or psychiatric management? AB - OBJECTIVE: To investigate the common perception that more serious suicide attempts tend to occur earlier in the day. METHODS: Prospective study of 158 adults referred for psychiatric assessment from the general hospital following an episode of deliberate self-harm. The main outcome measures used were Beck's Suicide Intent score, ICD-10 psychiatric diagnosis, alcohol consumption at the time of the attempt, and follow-up decision recorded by the interviewing duty psychiatrist. The patient also completed a checklist of current precipitating problems. RESULTS: A marked circadian variation in timing of the act was found, peaking between 2200 and 2400 h. "Early" acts (0300-1459 h) were significantly less likely to involve alcohol consumption, more likely to lead to admission to a medical ward, and involved more patient-identified problems than "late" acts. People who took overdoses early in the day were more likely to have concerns about their own mental health. Compared to earlier acts of self-harm, late evening (2200-2359 h) cases were less likely to be diagnosed as depressed or offered psychiatric follow up. No relation was found between time of day of self harm and Beck's Suicide Intent score. CONCLUSIONS: Implications arise regarding clinical risk assessment and current staffing levels in the accident and emergency department. The interviewing psychiatrist could concentrate on excluding depression and teaching problem solving to those who self-harm in the morning or afternoon, and on the detection and treatment of alcohol dependence for late evening cases. PMID- 11053598 TI - Organic syndromes diagnosed as conversion disorder: identification and frequency in a study of 85 patients. AB - BACKGROUND: The percentage of patients initially diagnosed with a conversion disorder and later identified as having an organic disorder has been decreasing in recent studies. METHOD: Consecutive patients with a diagnosis of conversion disorder were referred for psychiatric diagnosis and treatment. Research questions were: (1) What incidence of neurological disorder is revealed by neurological reassessment and by which diagnostic technique is the final diagnosis established? (2) What differences can be observed between true-positive and the false-positive results? RESULTS: Ten (11.8%) of the 85 patients examined appeared to suffer from a neurological disorder. In this sample, variables discriminating between the true positives and false positives were: (1) prior suspicion of neurological disorder; (2) older age at referral; (3) older age at onset of symptoms; (4) longer duration of symptoms; and (5) use of medication. Three variables contributed significantly to the prediction of organic disorder: prior suspicion of neurological disorder; age at onset of symptoms; and duration of symptoms. CONCLUSIONS: Although our results are in line with those of other recent studies, the percentage of false positives was still high. The data further emphasize the dangers of making a diagnosis of conversion disorder in the absence of positive evidence. It is important to continue to provide follow up for patients with a diagnosis of conversion disorder. Unfortunately, unreliable psychiatric indications, like certain behavioral characteristics, are still used in the diagnostic process. The results show that a general neurological examination is still a valuable diagnostic instrument in addition to modern diagnostic techniques. PMID- 11053599 TI - Men with prostate cancer: influence of psychological factors on informational needs and decision making. AB - OBJECTIVES: Studies indicate that men with prostate cancer (MPC) adopt passive roles in cancer management; however, increasing public awareness of prostate cancer and advocacy by MPC and their allies suggest otherwise. This study looks at the information that is important to MPC; their preferred participation in decision making; and the influence of sociodemographic, disease, and psychological factors on information needs and decision preferences. METHOD: Consecutive men diagnosed with prostate cancer and attending two tertiary care cancer clinics completed questionnaires on information needs and decision preferences. Questions included demographic information, health and disease status, psychosocial functioning, optimism, and decisional preferences and information preferences for content, type, focus, format, and amount. RESULTS: Questionnaires were completed by 101 MPC. Their mean age was 70 years and most were married and well-educated. Over 70% wanted detailed information at all illness stages focusing on their disease, treatment, survival, self-care, and empowerment. Over 60% of MPC wanted shared decision making with their physician. Psychological variables were found to influence information needs but not involvement in decision making. CONCLUSION: These results represent a challenge to health-care providers for accomodating the informational needs and decision preferences of individual MPC. PMID- 11053600 TI - The question of symptom lateralization in conversion disorder. AB - OBJECTIVE: The purpose of this study was to determine whether or not conversion symptoms are lateralized. Studies have shown a predominant left-oriented manifestation of symptoms for most somatoform disorders. The reports in the literature on the lateralization of conversion symptoms, however, are rather conflicting. They show left-sided, right-sided, or no symptom lateralization in conversion disorders. METHODS: One hundred fourteen patients with conversion disorder were screened for symptom lateralization. RESULTS: Those patients with unilateral symptoms (32.5%) showed no significant bias toward left or right symptom presentation. CONCLUSION: Based on these results, and the conflicting findings from previous studies, we conclude that there is insufficient support for lateralization theories in conversion disorder. PMID- 11053601 TI - Anxiety and depression in cancer patients: relation between the Hospital Anxiety and Depression Scale and the European Organization for Research and Treatment of Cancer Core Quality of Life Questionnaire. AB - BACKGROUND: The emotional functioning (EF) dimension of the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ C33) and the Hospital Anxiety and Depression Scale (HADS) evaluate anxiety and depression. We wanted to compare cancer patients' responses to EF with those to HADS, as well as the impact of anxiety and depression on the quality of life (QL) dimensions of the EORTC QLQ C33. METHOD: A total of 568 cancer patients completed both the EORTC QLQ C33 and HADS at the same occasion. The association between the patients' EF scorings and their HADS scores was analyzed by multiple linear regression. Gender and age were included as covariates. RESULTS: Statistically significant negative relations were found between EF and HADS-A (anxiety), HADS-D (depression) and HADS-T (total score), respectively, with the highest correlation coefficient for HADS-A. Older patients and males reported less emotional distress assessed by the EF scale than younger ones and females with comparable HADS-T or HADS-D scores. Both HADS-A and HADS-D were significantly related to other QL dimensions, and depression was a stronger predictor for reduced QL than anxiety. CONCLUSION: The EF dimension of EORTC QLQ C33 predominantly assesses anxiety, whereas depression is rated to a lesser degree. Combined with significant age and gender relations, this implies a risk of underdiagnosed depression, if the EORTC QLQ C33 is used as the only instrument to screen for psychological distress in cancer patients. As depression has a stronger impact on global QL of cancer patients than anxiety, the use of an additional instrument is recommended for assessment of depression. PMID- 11053602 TI - Positive and negative life events: the relationship with coronary heart disease risk factors in young adults. AB - OBJECTIVE: to investigate the influence of positive and negative life events (including daily uplifts and daily hassles) on several biological and lifestyle coronary heart disease (CHD) risk factors. METHODS: from the Amsterdam Growth and Health Longitudinal Study (AGHLS), a cross-section sample of 207 males and 231 females aged 32/33 years was used. RESULTS: hardly any associations were found between both positive and negative life events and biological CHD risk factors. On the other hand, daily uplifts and positive life events were positively related to lifestyle. For both positive and negative life events coping behaviour played a role in these relationships. Furthermore, it was shown that the associations of health-related variables with daily uplifts and hassles were comparable to those found for major positive and negative life events. CONCLUSION: This study could not fully determine whether or not different mechanisms play a role in the health benefits of positive life events compared to the health burdens of negative life events. PMID- 11053603 TI - Are alexithymia, depression, and anxiety distinct constructs in affective disorders? AB - OBJECTIVE: the present study was undertaken to gain a better insight into the relationship between alexithymia, anxiety, and depression. Two hypotheses were tested: (1) whether a depressive or anxiety disorder is associated with an elevation of one or more dimensions of alexithymia; and (2) whether alexithymia is an independent construct from depression and anxiety in patients with depressive or anxiety disorders. METHOD: a total of 113 patients with depressive or anxiety disorders (DSM-IV) and 113 control subjects completed the 20-item version of the Toronto alexithymia scale (TAS-20) and the hospital anxiety and depression scale (HADS). RESULTS: the TAS-20 total score was higher in depressed and anxious patients than in controls. This finding mainly depended on an increased score for "difficulty identifying feelings"(DIF), and (only in depressed patients) on an increased score for "difficulty communicating feelings" (DCF). The factor analysis of the TAS-20 and HADS items showed that depression is a construct different from alexithymia, whereas some overlap exists between anxiety and DIF dimension. CONCLUSION: our results suggest that in depressive and anxiety disorders, alexithymia and depression are separate constructs that may be closely related; in contrast, there are some overlaps between the DIF dimension and anxiety. PMID- 11053604 TI - Psychological and physical health correlates of body cell mass depletion among HIV+ men. AB - OBJECTIVE: To assess whether significant body cell mass depletion related to HIV is associated with declines in physical health and psychological well-being. METHODS: As part of a 2-year prospective HIV study, semiannual assessments included measures of body composition, psychological status, and physical health. RESULTS: As measured by bioelectric impedance analysis, 58 (31%) of 187 enrolled HIV+ men had significant body cell mass depletion at some point during the study, of who 23 subsequently lost at least an additional 5% of body cell mass in the 6 months between any two consecutive study visits. This additional body cell mass depletion was associated with significant increase in fatigue, global distress and depressive symptomatology, and reduced life satisfaction. CONCLUSION: These data illuminate the importance of monitoring body weight and body cell mass, and the need for awareness of the association between malnutrition, mental health, and quality of life. PMID- 11053605 TI - Negative affectivity, restriction of emotions, and site of metastases predict mortality in recurrent breast cancer. AB - OBJECTIVE: To assess whether negative affectivity and restriction of emotions predict survival time with recurrent breast cancer. METHODS: Thirty-two patients with recurrent breast cancer, diagnosed 6-19 months earlier and stabilized using surgical, medical and/or radiation therapies, were enrolled. Cox regression survival analyses, including initial severity of metastases (RR=4.3 [1.3-14.3]; p=0.02), were used to explore the association of psychological variables with survival. RESULTS: Low chronic anxiety in the context of low emotional constraint predicted low mortality (RR 0.07 [0.01-0.52]; p=0.007). However, patients with low chronic anxiety scores but with high constraint had higher mortality (RR=3.7 [1.2-11.5; p=0.02). High chronic anxiety, with or without high constraint, also predicted earlier death, as did high control of feelings. CONCLUSION: An integrated model of negative affectivity in the context of restriction of emotions appears to strengthen the prediction of survival based on severity of breast cancer metastases. PMID- 11053606 TI - Anger, depression, and coping interactions in headache activity and adjustment: a controlled study. AB - OBJECTIVE: To determine if individuals in the general community with chronic headache or migraine differ in terms of anger, depression, and coping strategies and from headache-free individuals in terms of anger and depression. METHOD: A community sample comprising 16 chronic tension-type headache (CTH), 28 migraine headache (MH), and 38 headache-free control subjects (CNT) were compared on measures of anger, depression, and use of various coping strategies. Affective and coping measures, recorded during a headache-free period, were regressed on headache activity measured in a daily diary over the following 2 weeks. Relationships between anger, depression, and coping were also examined in each of the headache groups. RESULTS: The MH subjects were found to use less effective coping strategies than controls and CTH, while the CTH group had higher levels on depression and various anger scales compared to controls and MH. Direct positive relationships were observed between suppressed anger and depression in the MH group, and between trait anger and depression in the CTH group. Anger and coping were predictive of headache activity in the following 2 weeks for both MH and CTH groups, while depression and coping, compared to coping only, were predictive of lifestyle interference from head pain in MH and CTH, respectively. CONCLUSION: The results support a relationship between affective and coping factors in headache activity and adjustment. PMID- 11053607 TI - Job strain, Type A behavior pattern, and the prevalence of coronary atherosclerosis in Japanese working men. AB - OBJECTIVE: To examine the relation of type A behavior pattern and job strain to angiographically documented coronary stenosis. METHODS: Subjects were 197 male Japanese patients with a full-time job. A questionnaire-based interview elicited psychosocial and other factors. Type A behavior pattern was measured by 12 questions, and job strain by the method of Karasek. Significant coronary stenosis was defined when a 75% or greater luminal narrowing occurred at one or more major coronary arteries or when a 50% or greater narrowing occurred at the left main artery. Logistic regression analysis was used to calculate odds ratio (OR) and 95% confidence interval (CI) with adjustment for traditional coronary risk factors and job type. RESULTS: Type A behavior pattern was related to a statistically non-significant lower prevalence of the coronary stenosis especially in the absence of job strain (adjusted OR 0.6, 95% CI 0.3-1.2). Job strain was non-significantly associated with a modestly increased prevalence of coronary stenosis (OR 1.7, 95% CI 0.6-5.2). CONCLUSION: These findings suggest that both the behavioral pattern and psychosocial work environment may be related to coronary artery stenosis. PMID- 11053608 TI - Attachment and loss experiences during childhood are associated with adult hostility, depression, and social support. AB - OBJECTIVES: This study examined developmental antecedents to psychosocial traits in adulthood that have been linked in prior studies to increased risk of heart disease. The hypothesis was tested that early parental loss coupled with poor quality family relationships (FR) during childhood would be associated with increased hostility and depression, and lower social support in adulthood. METHODS: Participants included 30 university students who experienced the death of one parent before the age of 16, and 31 control participants. Questionnaires were completed measuring current social support, hostility, depression, and the quality of FR. RESULTS: Multivariate analysis of variance (MANOVA) supported the hypothesis of maladaptive psychosocial characteristics in loss participants reporting poorer-quality FR. Significant interactions of loss and FR were found for individual variables of depressive symptoms, social support, and hostility. CONCLUSION: These results provide evidence that parental loss in childhood is associated with health-damaging psychosocial characteristics in adulthood only if the quality of the surviving FR is poor. PMID- 11053609 TI - Stress, coping, and depression in non-ulcer dyspepsia patients. AB - Thirty adults with upper gastrointestinal symptoms in the absence of structural organic disease diagnosed with non-ulcer dyspepsia (NUD) were compared to 30 healthy adults who had visited the hepatobiliary clinic for medical evaluation of non-organic complaints without NUD. Medical investigation in both groups were negative. Before independent gastrointestinal physicians conducted diagnostic evaluations, all subjects were evaluated for anxiety and depressive symptoms, stressful life events, coping style, and social support. The measures included Symptom Checklist 90-Revised (SCL-90-R), Beck Depression Inventory (BDI), Spielberger State-Trait Anxiety Inventory (STAI), Ways of Coping Checklist, and Interpersonal Support Evaluation List, and a self-report questionnaire, which measured the quantity of perceived stressful life events. The NUD patients reported significantly more symptoms of depression, more perceived stressful life events, less problem-focused coping, and less social support than the control subjects. Depressive symptoms were negatively correlated with interpersonal support, whereas, problem-focused coping was positively correlated with interpersonal support in the NUD patients. The two groups did not differ significantly in terms of anxiety and emotion-focused coping. The implications of these findings for the diagnosis and treatment of NUD are discussed. PMID- 11053610 TI - Thrombosis research 30 years. PMID- 11053611 TI - Plasma level of triglyceride-rich lipoprotein remnants is closely associated with the activation of coagulation factor VII in patients with myocardial infarction. AB - Remnant-like particles, which have been recognized to be atherogenic derivatives of chylomicrons and very low density lipoproteins, can be measured using a new assay kit. The purpose of the present study was to investigate the association of remnant-like particles with the coagulation system that has an important role in the pathogenesis of myocardial infarction. We assayed blood levels of total cholesterol, triglyceride, HDL-cholesterol, apolipoproteins, remnant-like particles-cholesterol, remnant-like particles-triglyceride, fibrinogen, factor VII antigen, activated factor VII, and tissue factor in 111 patients with a history of myocardial infarction and 128 control subjects. In simple regression analysis, plasma levels of remnant-like particles-cholesterol and remnant-like particles-triglyceride showed a significant positive correlation with the levels of activated factor VII (r=0.319, p<0. 001, and r=0.286, p=0.002, respectively) and the activated factor VII/factor VII antigen ratio (r=0.241, p=0.011, and r=0.249, p=0.008, respectively) in patients with myocardial infarction. In contrast, there were no significant differences between remnant-like particles and activated factor VII in control subjects. In stepwise multivariate regression analysis, the significant determinants of activated factor VII were remnant-like particles-cholesterol (10.2%), apolipoproteins A-I (5.1%), and E (7.1%); for the activated factor VII/factor VII antigen ratio, remnant-like particles triglyceride (6. 2%), age at blood sampling (5.1%), and apolipoprotein A-I (4.0%) in patients with myocardial infarction. However, the significant determinants of activated factor VII and the activated factor VII/factor VII antigen ratio were HDL-cholesterol (9.9% and 9.2%, respectively) in control subjects. It is concluded that remnant-like particles may be a risk factor for myocardial infarction by activating the extrinsic coagulation pathway. PMID- 11053612 TI - Hemostatic disturbances in patients with systemic inflammatory response syndrome (SIRS) and associated acute renal failure (ARF). AB - Endothelial damage plays a central role in the development of an SIRS-related Multiple Organ Dysfunction Syndrome (MODS) as a consequence of the establishment of a hemostatic imbalance between coagulation and fibrinolysis systems. Until now, sepsis is the SIRS model that has been most studied. The aim of this study was to assess the endothelial damage and the hemostatic imbalance in early stages of an SIRS of different origins, and to study if there are any differences in these disturbances between infectious and noninfectious SIRS. The endothelial damage and hemostatic changes were studied in 40 patients with SIRS (with less than 12 h of evolution) and an acute renal failure. Infectious SIRS was diagnosed in 19 cases and noninfectious SIRS in the remaining 21 patients. Patients with SIRS presented significantly higher values (p<0.001) for factors related to endothelial damage [von Willebrand factor (vWF), thrombomodulin, tissue plasminogen activator (t-PA), and plasminogen activator inhibitor type 1 (PAI-1) antigen], hypercoagulability [prothrombin fragment 1+2 (F1+2) and thrombin antithrombin complexes (TAT)], and fibrinolysis (D-dimer and PAI activity) with respect to the control group. However, although the group with infectious SIRS presented higher values for all the factors except for the t-PA and D-dimer with respect to SIRS of other origins, none of these differences reached statistical significance (p>0.05). Our data show that patients with SIRS and associated acute renal failure, irrespective of the origin (infectious or noninfectious), show signs of intense endothelial damage and hypercoagulability throughout the process. PMID- 11053613 TI - Platelet glycoprotein expression in patients with myelodysplastic syndrome. AB - Myelodysplastic syndromes (MDS) are characterized by a haematopoetic insufficiency that can lead to acute leukemia. A multistep pathogenesis caused by a clonal stem cell defect affecting several differentiation pathways has been proposed for MDS. Contrary to the better characterized alteration of lymphoid and myeloid differentiation, defects in thrombocytopoesis in MDS remain less clear. In the present study, we analyzed the expression of platelet glycoprotein (GP) Ia/IIa, IIb/IIIa, Ib/IX, and IV in 21 MDS patients (12 RA, 2 RARS, 4 RAEB, 1 RAEB T, 2 CMML) and healthy controls by flowcytometric analysis and quantitation of platelet GP RNA using fluorescence-based PCR. We observed a reduced cell surface expression of GPIb (p<0.01) and GPIIb/IIIa (p<0.01), while GPIa/IIa and GPIV expression was only marginally different between patients and controls. In contrast, there was a two-fold increase of platelet GPIb and GPIIb RNA and a three-fold increase of GPIV RNA among MDS patients. Increased levels of platelet GPIb and GPIIb RNA were significantly more prominent among patients with RAEB( T)/CMML (p<0. 05) in comparison to patients with RA/RARS. In conclusion, we demonstrate alterations in the cell surface expression and RNA content of platelet GPs in MDS patients. These data are consistent with dysmegakaryocytopoiesis and a defect in thrombocytopoiesis among MDS patients resulting from the clonal stem cell defect in MDS. PMID- 11053614 TI - The effect of vitamin C supplementation on coagulability and lipid levels in healthy male subjects. AB - Although dietary intake and plasma levels of vitamin C have been inversely associated with cardiovascular disease, the mechanism through which it may exert its effect has not been fully explained. Since thrombosis plays an important role in the onset of cardiovascular disease, we investigated the effect of vitamin C on measures of hemostasis that have been associated with cardiovascular risk. The effect of vitamin C on lipid levels was also evaluated. In a randomized, placebo controlled, crossover study, we determined the effect of 2 g daily of vitamin C supplementation on platelet adhesion and aggregation, levels of tissue plasminogen activator antigen, plasminogen activator inhibitor, fibrinogen, plasma viscosity, von Willebrand factor, and lipid levels in 18 healthy male volunteers with low normal vitamin C levels. No striking effects of vitamin C on the hemostatic measures were observed, although tissue plasminogen activator antigen levels were inversely related to Vitamin C levels. Von Willebrand factor levels were slightly higher with vitamin C, although within the normal range. Total cholesterol levels were 10% lower when subjects were receiving vitamin C compared to placebo (167+/-7 mg/dL vs. 184+/-7 mg/dL), P=0. 007), although the total cholesterol/HDL ratio was not significantly different. Higher levels of tissue plasminogen activator antigen, which in the present study were associated with lower vitamin C levels, have been shown in prospective studies to convey an increased risk of cardiovascular events. Further studies of the effect of vitamin C on hemostatic measures are required in higher risk populations or those with known cardiovascular disease. PMID- 11053615 TI - Angiotensin receptor antagonists during dextran sulfate LDL-apheresis are safe. PMID- 11053616 TI - Human anti-heparin-platelet factor 4 antibodies are capable of activating primate platelets: towards the development of a HIT model in primates. AB - In the first step to establish an animal model of heparin-induced thrombocytopenia (HIT) that is physiologically relevant to humans, studies were undertaken to determine the similarities or differences between human and non human primate (Macaca mulatta) platelets in HIT assay systems. The collagen-, ADP , and TRAP-induced platelet aggregation, and flow cytometric analysis of P selectin expression and microparticle formation were similar for both species platelets (p>0.1, n=18 each). The classical HIT assays using platelet-rich plasma (PRP) as well as a flow cytometric assay revealed the activation/aggregation and serotonin release assay (SRA) profiles for both primate and human platelets were similar in response to human HIT positive sera. All assays were heparin concentration-dependent; heparin, at 0.1 U/mL, produced maximum and similar platelet activation/aggregation and SRA responses with both primate (76+/-7%, n=18) and human (68+/-11%, n=20; p>0.1) platelets. At concentrations > or =10 U/mL, heparin suppressed the platelet aggregation and SRA responses in both systems. Primate and human platelets displayed similar behavior to low molecular weight heparin and pentasaccahride in HIT assay systems. Immunoglobulins isolated from serum of patients with HIT caused activation/aggregation of human (65+/-18%, n=10 donors) and primate (79+/-12%, n=6 monkeys, p>0.08) platelets. Unlike human platelets, the primate platelets exhibited a more consistent aggregation/release response (15 out of 18 primate platelets reactive). In contrast, human donors showed wide variations in the activation/release response (4 out of 10 reactive). These observations suggest that primate platelets are activatable by anti-H-PF4 antibodies, and support the hypothesis that primates can be used to develop an animal model to study the pathogenesis of HIT. PMID- 11053617 TI - Hirudin determination in plasma can be strongly influenced by the prothrombin level. AB - Recombinant hirudin is increasingly used for therapeutic and prophylactic anticoagulation. Several laboratory methods are available to measure r-hirudin, including clot-based and amidolytic methods. The snake venom ecarin converts prothrombin to meizothrombin. Hirudin inhibits meizothrombin, causing a prolongation of the ecarin clotting time (ECT). Because the ECT depends on prothrombin levels in plasma, it was compared with a chromogenic substrate assay (CSA) for the determination of r-hirudin levels in prothrombin deficient plasma samples. R-hirudin (0.0-2.0 microg/mL) was added to plasma samples with decreasing prothrombin concentrations (100-0%). Using the ECT, false high r hirudin levels were observed even in r-hirudin-free plasma, when prothrombin levels were below 50%. This effect was more pronounced with increasing r-hirudin levels. Additionally, r-hirudin (0.5 microg/mL) was added to plasma of patients with acquired prothrombin deficiency due to oral anticoagulation. Hirudin levels were also overestimated in these plasma samples using ECT. In plasma samples of patients treated with r-hirudin, because of suspected heparin-induced thrombocytopenia (HIT), hirudin levels were already measured falsely high, when the prothrombin levels were below 70%. The chromogenic substrate assay (CSA) determined correct values in all prothrombin-deficient plasma samples. Therefore, the CSA should be used for hirudin level determination, if overestimation due to prothrombin deficiency should be avoided. PMID- 11053618 TI - Activation of coagulation in C57BL/6 mice given verotoxin 2 (VT2) and the effect of co-administration of LPS with VT2. AB - To obtain better insight into the pathogenesis of verotoxin-producing Escherichia coli-associated diseases, in this study, we explored the effect of verotoxin 2 (VT2) on coagulation in an animal model. After being given VT2 (50 ng/kg, lethal dose), C57BL/6 mice showed progressively increasing expression of TF mRNA in the kidney and brain and elevated plasma levels of thrombin-antithrombin III complex (TAT), normotest, fibrinogen, and PAI-1 paralleling the disease course over 24 hours; platelet counts were decreased at 48 hours with hemorrhage in the kidney and brain. Co-administration of lipopolysaccharide (LPS, 0.5 mg/kg) with VT2 (50 ng/kg) exhibited more prominant and/or prolonged increase in not only expression of TF and PAI-1 mRNAs in the kidney and brain but also plasma levels of TAT, fibrinogen, and PAI-1 and was associated with more remarkable hemorrhage in the tissues. Although VT2 (5 ng/kg) was not a lethal dose, co-administration of LPS (0.5 mg/kg) with VT2 (5 ng/kg) enhanced the susceptibility to VT2, resulting in more prolonged elevation of TAT levels during the first 24 hours than that in the LPS group and a second elevation at 72 hours, followed by death. Plasma IL-1beta level reached a maximum at 24 hours after VT2 (50 ng/kg) injection prior to the increase in TAT levels, whereas the increase in TNFalpha level immediately after injection was associated with the increase in PAI-1 mRNA. These observations indicate that the activation of coagulation by VT2 may occur through a mechanism different from that used by LPS, since plasma TAT levels rose in the mice immediately after LPS injection and returned to normal over 36 hours. PMID- 11053619 TI - Comparing the antithrombotic efficacy of a humanized anti-factor IX(a) monoclonal antibody (SB 249417) to the low molecular weight heparin enoxaparin in a rat model of arterial thrombosis. AB - A humanized inhibitory anti-factor IX(a) antibody (SB 249417) has been compared to enoxaparin (Lovenox) in a rat model of arterial thrombosis. Pretreatment of rats with either SB 249417 (3.0 mg/kg, i. v.) or enoxaparin (30.0 mg/kg, i.v. or s.c.) resulted in comparable and significant reductions in thrombus formation. However, the efficacious dose of enoxaparin resulted in >30-fold increase in the aPTT over baseline, while the efficacious dose of SB 249417 prolonged the aPTT by only approximately 3-fold. Additionally, pretreatment with SB 249417 resulted in sustained blood flow and arterial patency throughout the experiment in >80% of rats treated. In contrast, <30% of rats pretreated with enoxaparin remained patent throughout the experiment. The data in this report indicate that the selective inhibition of factor IX(a) with the monoclonal antibody SB 249417 produces a superior antithrombotic profile to that of the low molecular weight heparin enoxaparin. PMID- 11053620 TI - Expression and characterization of recombinant protein S with the Ser 460 Pro mutation. AB - To characterize the putative biochemical modifications induced by the Ser 460 to Pro (Heerlen) mutation in protein S (PS), we expressed both wild-type (wt) and mutated recombinant PS in HEK cells. In SDS-polyacrylamide gels, r-PS Heerlen migrated at 71 kDa whereas r-wt PS migrated at 73 kDa, a difference abolished after deglycosylation by N-glycosidase, suggesting that the Ser 460 Pro mutation abolishes N-glycosylation of Asn 458. The affinity of r-wt PS and r-PS Heerlen for C4b-binding protein (C4b-BP) and for phospholipid vesicles was similar. Neither the enhancement of APC-dependent prolongation of the APTT, nor the specific enhancement of FVa and FVIIIa proteolysis by APC in purified systems was affected by the mutation. However, the Ser 460 Pro mutation induced a slight conformational change in the SHBG domain of the PS molecule, as shown by reduced binding affinity for monoclonal antibodies. The type III phenotype associated with the Heerlen mutation might thus result from a slightly modified rate of synthesis or catabolism. The resulting moderate decrease in the circulating PS concentration may modify the equilibrium between free PS and C4b-BP/PS complexes. PMID- 11053621 TI - Inhibition of arterial thrombosis by a peptide ligand of the thrombin receptor. AB - Thrombin plays an important role in promoting arterial thrombosis by platelet activation and by catalyzing fibrin formation. Use of thrombin inhibitors that block both the platelet-activating and fibrin formation properties of thrombin are associated with hemostasis. This problem might be overcome by developing agents that block only the platelet-activating property of thrombin. Because the platelet-activating property of thrombin is mediated by the thrombin receptor, antagonists of the thrombin receptor might be efficacious and potentially safer with regard to bleeding complications. We investigated whether a peptide ligand (AFLARAA) of the thrombin receptor that blocked alpha-thrombin and thrombin receptor activating peptide-induced platelet aggregation could inhibit thrombosis. A partially occlusive thrombus was generated by application of electric current in rabbit carotid artery. In control animals, the artery was completely occluded within 42+/-12 min after the current was discontinued. When the thrombin receptor activating peptide antagonist was given (100 micromol/kg as an IV bolus followed by 900 micromol/kg infusion for a period of 180 min) starting at the time the current was stopped, blood flow remained patent throughout the infusion period and for an additional 60 min after the infusion was stopped. The antithrombotic effect of the antagonist peptide was not associated with increased bleeding tendency, as judged by the amount of blood adsorbed by a gauze pad placed in a surgical incision extending to the muscle tissue and by a standard template bleeding time. These results indicate that thrombin receptor antagonist peptides can be used as antithrombotic agents. PMID- 11053622 TI - In vitro and in vivo studies of AT-1362, a newly synthesized and orally active inhibitor of thrombin. AB - AT-1362 was found to be a potent, selective, and competitive inhibitor of thrombin, with a Ki value of 6.7 nM. In a rat model of venous thrombosis induced by partial stasis and endothelial disruption, the ID(50) values (a dose required to obtain 50% inhibition of thrombus formation over each vehicle group) of AT 1362 and argatroban were 0.03 mg/kg i.v. plus 0.5 microg/kg/minute and 0. 13 mg/kg i.v. plus 8.7 microg/kg/minute, respectively, and the antithrombotic effect of AT-1362 without prolongation of bleeding time lasted for 2 hours and disappeared 4 hours after oral administration of 30 mg/kg. In the rat tail transection model, the BT(2) values (a dose causing two-fold prolongation of the bleeding time over each vehicle group) of AT-1362 and argatroban were 0.56 mg/kg i.v. plus 9.3 microg/kg/minute and 1.1 mg/kg i.v. plus 73.3 microg/kg/minute, respectively. The reduction of thrombus formation and the prolongation of bleeding time were correlated with an ex vivo activated partial thromboplastin time (APTT) for both drugs. AT-1362 at 0.3 mg/kg i.v. plus 5 microg/kg/minute and argatroban at 0.6 mg/kg i.v. plus 40 microg/kg/minute significantly (p<0.05 and p<0.01, respectively) improved the vessel patency in a FeCl(2)-induced carotid artery thrombosis model in rats. These results suggest that AT-1362 may be a potent antithrombotic agent for the treatment of thrombotic diseases. PMID- 11053623 TI - A novel type of mutation at the propeptide cleavage site (AlA+1Thr) causing symptomatic protein C type II deficiency. PMID- 11053624 TI - Low molecular weight heparins: are they superior to unfractionated heparins to prevent and to treat deep vein thrombosis? AB - In many countries, low molecular weight heparins (LMWHs) have replaced unfractionated heparin (UH) for prevention and treatment of venous thromboembolism. The present paper reviews the possible advantages of LMWHs over UH. In spite of their lower molecular weight distribution, LMWHs are functionally more heterogeneous than UH. Their anti-Xa/anti-IIa ratio varies significantly, and the injection of the same dose generates different anti-Xa activities and activated partial thromboplastin time (APTT) prolongations. Their pharmacodynamic properties account for their more convenient use in comparison with UH; however, there is a risk of accumulation in case of renal insufficiency. Even if they are less anticoagulant on the basis of the APTT prolongation, they are not less prohemorrhagic than UH. LMWHs are probably less immunogenic and probably induce less osteoporosis. Several meta-analyses published between 1992 and 1999 indicate that LMWHs are as efficient as UH in preventing postoperative deep vein thrombosis (DVT) in general surgery and more efficient than UH in preventing DVT in orthopedic surgery and treating established DVT. PMID- 11053625 TI - For the initial treatment of venous thromboembolism: are all low-molecular-weight heparin compounds the same? AB - Low-molecular-weight heparin compounds have been used in the treatment of patients with venous thromboembolism for approximately 15 years. Ever since their introduction, there has been discussion about whether low-molecular-weight heparin compounds differ in their efficacy and safety. The best answer would be provided by direct comparison of different low-molecular-weight heparin preparations; however, these trials have not been conducted. Classical meta analysis has its limitations for such a comparison since only a very small number of trials with the respective low-molecular-weight heparin compounds are available. The objective of the present analysis has been the use of meta regression to compare the efficacy and safety of different low-molecular-weight heparin compounds in the initial treatment of patients with venous thromboembolism. We used computerized literature searches to identify studies that compared dose-adjusted unfractionated heparin treatment with fixed dose subcutaneous low-molecular-weight heparin treatment in patients with established venous thromboembolism. The individual odds ratios of the studies were plotted against the absolute percentage of the major outcomes in the unfractionated heparin control group. Linear regression was used to find differences between different low-molecular-weight heparin compounds. There appears to be some variation in efficacy and safety among the currently available low-molecular weight heparin preparations. PMID- 11053626 TI - The role of low-molecular-weight heparins in arterial diseases: optimizing antithrombotic therapy. AB - On the basis of current evidence, all patients with acute coronary syndromes should receive optimized medical therapy, whether or not they ultimately undergo an invasive revascularization procedure, to improve both clinical outcomes and cost effectiveness. While standard aspirin and unfractionated heparin (UFH) have improved short-term outcomes, they do not eliminate the risk of recurrent ischemic episodes. The recent introduction of platelet fibrinogen receptor antagonists and low-molecular-weight heparins (LMWHs) has offered an opportunity to develop more aggressive antithrombotic regimens. The LMWHs have been thoroughly evaluated in unstable angina and non-Q wave myocardial infarction (UA/NQMI), and have demonstrated improved efficacy compared to standard UFH, without an increase in major complications caused by bleeding. Experience has also been gathered using LMWHs in other arterial diseases (such as pregnant patients with prosthetic heart valves) and as an adjunctive therapy with thrombolytics for acute myocardial infarction. Lastly, studies are currently underway evaluating LMWHs in patients with atrial fibrillation. PMID- 11053627 TI - Comparison of in vitro immunostimulatory potential of live and inactivated influenza viruses. AB - Live influenza viruses, heat-inactivated virus, and a trivalent formalin inactivated influenza vaccine were analyzed for their in vitro stimulatory properties on immune cells from healthy donors. Lymphocyte proliferation induced by each influenza antigen was comparable. Influenza vaccine stimulated significantly lower production of interferon-gamma (IFN-gamma) compared with live and heat inactivated viruses, whereas both vaccine and heat-inactivated influenza induced lower levels of IFN-alpha compared with live virus. Furthermore, only live virus generated influenza-specific cytotoxic T lymphocyte (CTL) activity. A significant increase in monocyte expression of CD80, CD86, CD40, and human leukocyte antigen-DR (HLA-DR) was also induced by live influenza virus. Our results suggest that immunization with live influenza vaccines might induce immune responses that would not be induced by conventional inactivated vaccines, including CTL generation, antiviral IFN-gamma and IFN-alpha cytokine production, and increased antigen presentation and costimulatory capacity on antigen presenting cells (APC). PMID- 11053628 TI - Lack of CIITA expression is central to the absence of antigen presentation functions of trophoblast cells and is caused by methylation of the IFN-gamma inducible promoter (PIV) of CIITA. AB - Lack of MHC-mediated antigen presenting functions of fetal trophoblast cells is an important mechanism to evade maternal immune recognition. In this study we demonstrated that the deficiency in MHC expression and antigen presentation in the trophoblast cell lines JEG-3 and JAR is caused by lack of class II transactivator (CIITA) expression due to hypermethylation of its interferon-gamma (IFN-gamma)-responsive promoter (PIV). Circumvention of this lack of CIITA expression by introduction of exogenous CIITA induced cell surface expression of HLA-DR, -DP, and -DQ, leading to an acquired capacity to present antigen to antigen-specific T cells. Transfection of CIITA in JEG-3 cells also upregulated functional HLA-B and HLA-C expression. Noteworthy, this lack of IFN-gamma mediated induction of CIITA was also found to exist in normal trophoblast cells expanded from chorionic villus biopsies. Together, these observations demonstrate that lack of CIITA expression is central to the absence of antigen presentation functions of trophoblast cells. PMID- 11053629 TI - A single nucleotide polymorphism in the first intron of the human IFN-gamma gene: absolute correlation with a polymorphic CA microsatellite marker of high IFN gamma production. AB - We have described previously a variable length CA repeat sequence in the first intron of the human IFN-gamma gene and showed that allele #2 is associated with high in vitro IFN-gamma production. In a consecutive study, allele #2 was found to be associated with allograft fibrosis in lung transplant patients, confirming its role as a marker of high IFN-gamma production, both in vivo and in vitro. In this study we have sequenced 50 PCR products that had been typed previously by PAGE for the identification of CA microsatellite alleles. We report on a novel single nucleotide polymorphism, T to A, at the 5' end of the CA repeat region in the first intron of the human IFN-gamma gene (+874*T/A). There is an absolute correlation between the presence of T allele and the presence of the high producing microsatellite allele #2. This T to A polymorphism coincides with a putative NF-kappa B binding site which might have functional consequences for the transcription of the human IFN-gamma gene. Therefore, the T to A polymorphism could directly influence the level of IFN-gamma production associated with the CA microsatellite marker. PMID- 11053630 TI - A 70 kDa MHC class I associated protein (MAP-70) identified as a receptor molecule for Coxsackievirus A9 cell attachment. AB - One of the major categories of disease-causing micro-organisms are viruses. New studies on many different viruses have shown that virus attachment and cell entry is often a multistep process, requiring many interactions between the virus and cell surface molecules. In this study, we have attempted to identify the cell surface molecules involved in Coxsackievirus A9 (CAV-9), a common human pathogen and a member of the Picornavirus family, infectious process. GMK cells susceptible to virus infection were surfaced labeled with biotin and then solubilized in non-ionic and zwiterionic detergents. Free CAV-9 virions were used as an affinity surface, allowing the virus to bind to the solubilized receptors. The virus-receptor complexes were then immunoprecipitated by an anti CAV-9 serum and protein-A sepharose beads. SDS-PAGE and two-dimensional electrophoresis revealed the presence of integrin alpha v beta 3 molecules and a 70 kDa protein with apparent isoelectric point (pI) 5.5. The identity of the integrin alpha v beta 3 molecules was confirmed by immunoprecipitation and Western blotting; whereas the 70 kDa protein was also found to co-immunoprecipitate with MHC class I molecules in non-stringent conditions. Sequential immunoprecipitation experiments confirmed that the MHC class I associated protein (MAP-70) and the 70 kDa protein utilized by CAV-9 were identical. The role of MAP-70 in CAV-9 infectious process is discussed. PMID- 11053631 TI - In vitro CTL precursor frequencies do not reflect a beneficial effect of cross reactive group (CREG) matching. AB - Adjustment of histocompatibility-based allocation criteria in kidney transplantation from HLA matching to matching on the basis of cross-reactive groups (CREG), was recently suggested to be a good alternative to transplant with more "well-matched" kidneys, without negatively influencing graft survival. Because graft rejection is often mediated by cytotoxic T cells (CTLs), we investigated whether a beneficial effect of CREG matching is reflected in vitro by lower CTL precursor frequencies (CTLpf). Therefore, CTLpf were determined in a group of healthy individuals and analyzed with respect to the number of HLA and CREG mismatches. A clear correlation was found between the number of HLA mismatches and the CTLpf, that is, the lowest mean frequency in case of 0 HLA-A, B mismatches (66 CTL precursors per 10(6) cells) and the highest in combinations with 4 HLA mismatches (mean = 303 CTLp/10(6) cells). The situation was different in the case of CREG mismatches. Although the highest frequency was found in the group of 4 CREG mismatches, no significant differences were observed between 0, 1, and 2 CREG mismatches. High CTLpf, up to 430/10(6), were even seen in the case of 0 CREG mismatches. Also within a well-defined group of single HLA-A or HLA-B mismatches no difference in CTLpf were observed between the subgroups with 0 vs. 1 CREG mismatches. The present study showed that in vitro the CTLpf correlates better with HLA than with CREG matching. These data are consistent with findings reported by several groups that matching for the CREG does not benefit transplant outcome. PMID- 11053632 TI - Switch from cyclosporine A to tacrolimus in renal transplant recipients: impact on Th1, Th2, and monokine responses. AB - We showed previously that pretransplant CD4 helper defects and low in-vitro IL-10 responses predict a low risk of acute kidney graft rejection. To compare the effect of tacrolimus (Tacr) and cyclosporine A (CsA) on the humoral immune response we assessed T helper function, B cell/monocyte responses and in-vitro cytokine responses (TNF-alpha, GM-CSF, IL-1 beta, IL-2, IL-4, IL-6, IL-10) in 20 renal transplant recipients before and 3 months after they were switched from CsA to Tacr because of hyperlipoproteinemia, hirsutism, or gum hyperplasia. T helper function was assessed using a PWM-driven allogeneic coculture system of patient T cells together with control B cells. B cell/monocyte responses were determined using a PWM-stimulated allogeneic coculture system, SAC I-stimulated B-cell cultures and LPS-stimulated monocyte cultures. Immunoglobulin-secreting cell (ISC) responses were assessed in a reverse hemolytic plaque assay, and ELISA were used to determine cytokine secretion. Treatment with Tacr resulted in a decreased expression of costimulatory ligands and adhesion molecules (T cells: CD40L, p < 0.05; CD28 and CD54, p < or = 0.01; B cells: CD25, p = 0.05; CD40, p < 0.001; monocytes: CD40, p < 0.05), which coincided with decreased PHA-stimulated T cell IL-2 responses (398 +/- 153 versus 43 +/- 15 pg/ml, p < 0.05), impaired CD4 helper activity (117% +/- 22% versus 73% +/- 19%, p < 0.05) and increased CD4 suppressor activity (-120% +/- 28% versus -18% +/- 27%, p = 0.02). We observed enhanced CD4 IL-10 responses (p < 0.01) and LPS-stimulated monocyte responses (TNF-alpha, IL-1 beta, and IL-6, p < 0.005; IL-10, p < 0.05), indicating an increased humoral immune responsiveness under treatment with tacrolimus. Our data show that switching of immunosuppressive therapy from CsA to tacrolimus results in suppression of costimulatory ligands, adhesion molecules, Th1 responses and CD4 helper activity. However, enhanced humoral immune responses, Th2 and monokine responses, might have a negative impact on long-term graft function. PMID- 11053633 TI - T cell immune reconstitution after allogeneic bone marrow transplantation in bare lymphocyte syndrome. AB - To study the impact of an MHC class II-negative environment on T cell immune reconstitution, we have analyzed the phenotypical and functional characteristics of FACS-sorted cultured CD4(+) and CD8(+) T cells in two Bare Lymphocyte Syndrome (BLS) patients before and after allo-BMT. A similar analysis was performed in two MHC class II expressing pediatric leukemia patients after treatment with an allo BMT who were included in our study as control. It was observed that CD4(+) T cells displayed cytolytic alloreactivity in both BLS patients prior to and within the first year after allo-BMT, whereas such cells were absent at a later time point, in the donors and pediatric leukemia controls. In addition, reduced MHC class II expression was observed in CD8(+) T cells of both recipients early after allo-BMT, irrespective of the T cell chimerism pattern. Lack of endogenous MHC class II expression in BLS patients, therefore, results in aberrant T cell selection within the first year after allo-BMT, analogous to T cell selection before transplantation. These T cell selection processes seem to be normalized at a later time point after allo-BMT probably due to migration and integration of graft-derived MHC class II-positive antigen presenting cells to sites of T cell selection. PMID- 11053634 TI - Functional alteration of granulocytes, NK cells, and natural killer T cells in centenarians. AB - The immune system in centenarians was characterized as elevated levels in the proportion and number of granulocytes, NK cells, and extrathymic T cells (including NKT cells) in the peripheral blood. Conventional T cells, abundant in youth, were decreased in proportion and number. In addition to this numerical change in centenarians, the function was significantly altered in comparison with that in middle-aged subjects. The phagocytic function and cytokine production of granulocytes in centenarians increased whereas the production of superoxides from granulocytes decreased. This tendency was almost the same in both healthy and unhealthy centenarians. IFN gamma production by NK and extrathymic T cells in centenarians seemed to be augmented and resulted in an elevated level of serum IFN gamma. Possibly due to the effect of this endogenous IFN gamma, the proportion of CD64(+) (Fc gamma RI) cells among granulocytes was elevated. The expansion of CD64 antigens on granulocytes is known to be regulated by IFN gamma and to be associated with their induction of phagocytosis. These results suggest that the immune system of centenarians is not merely impaired, but altered in terms of the number and functions of granulocytes, NK cells, NKT cells. PMID- 11053635 TI - Identification of MICA as a new polymorphic alloantigen recognized by antibodies in sera of organ transplant recipients. AB - MHC class I-related chain A (MICA) is an HLA-related, polymorphic gene the product of which may be recognized by a subpopulation of intestinal gamma delta T cells and may play a role in the activation of a subpopulation of natural killer cells. Using anti-MICA specific rabbit sera we previously demonstrated that freshly isolated monocytes, keratinocytes, fibroblasts, and endothelial cells express MICA. To analyze whether MICA may be a target for specific antibodies in sera of transplanted patients, we produced three recombinant MICA proteins consisting of the alpha 1, alpha 2, and alpha 3 domains, and used them in an enzyme-linked immunosorbent assay. We found that several patients had specific antibodies against MICA. Most of them were detected in serum samples collected at different times after organ rejection. Although this finding raises the question of how these patients became immunized, the fact that the polymorphic, HLA-like MICA molecule, expressed at the cell surface of endothelial cells, is recognized by specific antibodies in sera of transplanted patients, suggests the MICA may be a target molecule in allograft rejection. PMID- 11053636 TI - HLA-C(*)03 is a risk factor for cardiomyopathy in Chagas disease. AB - Previous studies have shown the effect of class 1 as detected by serology or class 2 HLA genes by oligotyping upon susceptibility or resistance to the cardiomyopathy that develops in approximately one third of the Trypanosoma cruzi chronically infected patients. Low and intermediate resolution DNA typing of class 1 alleles was performed in a sample of 113 serologically positive individuals with and without cardiomyopathy. A polymerase chain reaction-sequence specific oligonucleotide probe method using primers and probes from the British Society of Histocompatibility and Immunogenetics as modified for the VII Latin American Histocompatibility Workshop by D. Middleton, and LiPA kits from Innogenetics were used. Several alleles (A(*)11, A(*)31, B(*)15, B(*)35, B(*)45, B(*)49, B(*)51, and C(*)03) showed increased frequencies among patients with cardiac damage versus the asymptomatic group, but only the last one remained significant after correction of the p value (OR = 5.8, p(c) = 0.03). HLA-C(*)03 showed linkage disequilibrium with B(*)40 and B(*)15 and although both haplotypes were increased in cardiopathic patients compared with asymptomatic individuals, the difference is not significant. These results suggest that the HLA-C*03 allele could confer susceptibility to the development of cardiomyopathy among Venezuelan T. cruzi seropositive individuals and contrast with the protective effect conferred by the HLA B40 Cw3 haplotype among Chilean chagasic patients. Further studies will be needed to confirm the role of this allele on the cardiomyopathy of Chagas disease. PMID- 11053637 TI - HLA-DRB1 and DQB1 polymorphisms in southern France and genetic relationships with other Mediterranean populations. AB - This study presents the results of HLA-DRB1 and DQB1 sequence-specific oligonucleotide probe (SSOP) typing for a population sample of 181 individuals originating from southern France. On the basis of allele and haplotype frequencies, we compared our population with others from the Mediterranean area. Allele frequencies are comparable to those found in other western European populations (France, Portugal, Spain) and indicate neighboring exchanges. The haplotype frequencies showed relationships with North Africans and Jewish populations, as well as the common origin of Moroccan and Lebanese Jews. Therefore, allele frequencies seem to be more able to show recent exchanges while haplotype frequencies might show ancestral relationships. These results may serve as references for future studies of HLA and disease in southern France. PMID- 11053638 TI - HLA-DR2-associated DRB1 and DRB5 alleles and haplotypes in Koreans. AB - There are considerable racial differences in the distribution of HLA-DR2 associated DRB1 and DRB5 alleles and the characteristics of linkage disequilibrium between these alleles. In this study, the frequencies of DR2 associated DRB1 and DRB5 alleles and related haplotypes were analyzed in 186 DR2 positive individuals out of 800 normal Koreans registered for unrelated bone marrow donors. HLA class I antigen typing was performed by the serological method and DRB1 and DRB5 genotyping by the PCR-single strand conformational polymorphism method. Only 3 alleles were detected for DR2-associated DRB1 and DRB5 genes, respectively: DRB1(*)1501 (gene frequency 8.0%), (*)1502 (3.2%), (*)1602 (0.9%); DRB5(*)0101 (8.0%), (*)0102 (3.2%), and (*)0202 (0.9%). DRB1-DRB5 haplotype analysis showed an exclusive association between these alleles: DRB1*1501 DRB5*0101 (haplotype frequency 8.0%), DRB1(*)1502-DRB5(*)0102 (3.2%), and DRB1(*)1602-DRB5(*)0202 (0.9%). The 5 most common DR2-associated A-B-DRB1 haplotypes occurring at frequencies of > or = 0.5% were A24-B52-DRB1(*)1502 (1.8%), A2-B62-DRB1(*)1501, A2-B54-DRB1(*)1501, A26-B61-DRB1(*)1501, and A24-B51 DRB1(*)1501. The remarkable homogeneity in the haplotypic associations between DR2-associated DRB1 and DRB5 alleles in Koreans would be advantageous for organ transplantation compared with other ethnic groups showing considerable heterogeneity in the distribution of DRB1-DRB5 haplotypes. PMID- 11053639 TI - HLA, aging, and longevity: a critical reappraisal. AB - Despite a large number of studies, available data do not allow at present to reach definitive and clear conclusions on role of HLA on longevity, owing to major methodological problems, such as serological and molecular typing of different loci, insufficient sample sizes, different inclusion criteria and age cut-off, inappropriate mixing of data referred to people from 58 to over 100 years of age, inappropriate control matching, and neglected consideration of sex related effects and the different genetic make-up of studied populations. However, within this confused scenario, some data emerge. First, two studies that do not fit the biases above discussed show that some HLA alleles are associated with longevity. However, some of these alleles may confer an increased risk to undergo a variety of diseases. Second, longevity may be associated with an increased homozygosity at HLA loci. Third, an intriguing association between longevity and the 8.1 ancestral haplotype (AH), which has been proven to be associated with a variety of immune dysfunctions and autoimmune diseases, apparently emerges. This association appears to be a sex-specific (males) longevity contributor, and it is particularly interesting, taking into account that a type 2 (early infancy) --> type 1 (adulthood) --> type 2 (aging) shift of cytokine profile occurs lifelong, and that individuals bearing this haplotype show a type 2 immune responsiveness (note that type 1 cytokines mainly enhance cellular responses, whereas type 2 cytokines predominantly enhance humoral responses). On the whole, the (sex specific) association of longevity with alleles or haplotypes of several genes related to risk factors for a variety of diseases (cardiovascular diseases, cancer), including HLA alleles and haplotypes, is not unexpected on the basis of previous studies on the genetics of longevity in centenarians. This association can be interpreted under the perspective of a well known evolutionary theory of aging (antagonistic pleiotropy). This theory predicts that the same gene (or allele or haplotype) can have different roles (positive or negative) in different periods of the life span. Thus, the 8.1 AH should exert a positive effect during the infancy and aging but not in adulthood, when, indeed it is associated to susceptibility to a variety of diseases. PMID- 11053640 TI - Nomenclature for factors of the HLA system, update June 2000. WHO Nomenclature Committee for Factors of the HLA system. PMID- 11053641 TI - Iron, alpha-tocopherol, oxidative damage and micronucleus formation in rat splenocytes. AB - The influence of low and high alpha-tocopherol diets in concert with a high polyunsaturated fat content and a modest increase in dietary iron has been studied. Iron supplementation at five times the recommended dietary level was not associated with any increased sensitivity of the splenocytes to any of the oxidative challenges. Despite the significantly higher alpha-tocopherol concentrations in the plasma and liver of animals supplemented with this vitamin, there was no apparent protection against oxidative genotoxicity, as judged by the formation of micronuclei in splenocytes subjected to oxidative stress ex vivo. These results add to the evidence that vitamin E supplementation has little effect against oxidative genomic damage, at least as demonstrated by an increase in micronucleus frequency. PMID- 11053642 TI - In vivo gene expression profile analysis of metallothionein in renal cell carcinoma. AB - The antiapoptotic and mitogenic responses of metallothionein (MT) have been well documented in vitro. While MT protein overexpression, frequently encountered in a number of human primary tumors, has been shown to be correlated with disease progression, little information is available on the in vivo isoform expression of MT. In this study we have demonstrated the occurrence of MT proteins and further defined their differential expression profile in human primary renal cell carcinoma (RCC). Pooled normal human kidney RNA and paired biopsy specimens (tumor and control) obtained from 11 patients diagnosed with RCC with tumor grade ranging from 1-3 and a pathological staging of T2-T3 (N0M0) were used for the study. Samples were analyzed for the presence of MT protein using immunohistochemical (IHC) analysis and for MT isoform-specific mRNA expression by reverse transcriptase polymerase chain reaction. Metallothionein protein assumed both cytoplasmic and nuclear staining in cancer cells and was detected in eight of 11 samples (72%) with polyclonal antibodies. The immunoreactivity of MT protein, but not its cellular localization, in RCC specimens suggests a relationship between and advanced disease. While alterations in the basal level of expression of MT-1E, MT-1F and MT-1X genes remained unchanged, significant up regulation of MT-2A and down-regulation of MT-1A and MT-1G transcripts was observed in RCC tissue specimens when compared with controls. Intriguingly, the paired RCC biopsy specimens had lower MT-1H transcripts than pooled normal human controls. We here provide the first report of the differential expression of MT isoforms in human RCC and that this data further support the role of MT-2A in tumorigenesis. PMID- 11053643 TI - Modifying effects of 4-phenylbutyl isothiocyanate on N-nitrosobis(2 oxopropyl)amine-induced tumorigenesis in hamsters. AB - The modifying effects of dietary 4-phenylbutyl isothiocyanate (PBITC), given during the initiation stage of carcinogenesis, were investigated in hamsters treated with N-nitrosobis(2-oxopropyl)amine (BOP). A total of 120 female 5-week old hamsters were divided into six groups. Animals in groups 1-3, each consisting of 30 hamsters, were given BOP by two subcutaneous injections, 1 week apart, at a dose of 20 mg/kg body weight, plus 0, 10 or 100 micromol/animal of PBITC in corn oil by gavage 2 h prior to each carcinogen treatment. Ten animals in group 4 served as a vehicle control, and animals in groups 5 and 6, each consisting of ten hamsters, were given 10 and 100 micromol of PBITC alone in corn oil. Sacrifice was 52 weeks after the first BOP injection. The PBITC treatments significantly (P<0.05) inhibited the development of pancreatic ductal dysplasias and adenocarcinomas. Also, lung tumors (adenomas and adenocarcinomas) were significantly (P<0.05) reduced in a dose-dependent manner. In contrast, both hepatocellular and cholangiocellular tumors (adenomas and carcinomas) tended to be or were significantly increased by PBITC. These results, taken together with our previous findings, indicate that the natural isothiocyanate, phenethyl isothiocyanate (PEITC), has a more potent chemopreventive action against BOP induced tumorigenesis than synthetic isothiocyanates with longer alkyl chains, such as 3-phenylpropyl isothiocyanate (PPITC) and PBITC. Thus, their lipophilicity does not necessarily reflect the chemopreventive potential because the strength of lipophilicity is PEITCMFII-->MFIII) in the precursor compartment and three types of postmitotic fibrocytes (PMFIV-->PMFV-->PMFVI) in the functional compartment has been identified previously. In the present study, we show that replenishment of fibrocytes lost from the functional compartment is not expected to change the distribution of differentiation types in a steady state population, provided cell loss occurs at the end of a long sequence of cell divisions only. However, premature terminal differentiation of progenitor fibroblasts to postmitotic fibrocytes can be induced by ionising radiation and other cell stressors. Furthermore, even a low dose of 1Gy causes a change in the distribution of surviving MF progenitor cells towards later differentiation stages within the precursor compartment. The role of autocrine transforming growth factor-beta1 production by fibroblasts in mediating terminal differentiation was investigated. We propose that cell stress and DNA damaging agents may contribute to progression of the differentiation state with age and that individual variation may be related to differences in the rate of induced differentiation. PMID- 11053666 TI - Comparison of the protective activities generated by two survival proteins: Bcl-2 and Hsp27 in L929 murine fibroblasts exposed to menadione or staurosporine. AB - Hsp27 and Bcl-2 are survival proteins that protect against cell death. We have compared the specific protective activity (protection per number of molecules expressed) mediated by these proteins when they are expressed in L929 murine fibroblasts. We found that Hsp27 and Bcl-2 efficiently delayed the cytotoxicity generated by menadione. Both proteins interfered with the mitochondria membrane potential collapse, the reactive oxygen species (ROS) burst and the decrease in glutathione level induced by this oxidant. In untreated cells, both proteins decreased the ROS levels and raised the glutathione cellular content. Taking their levels of expression into account, we concluded that Bcl-2 was much more active than Hsp27 for counteracting the above-mentioned menadione effects, and for modulating the ROS and glutathione levels in untreated cells. Both Hsp27 and Bcl-2 also conferred cellular resistance to staurosporine, a kinase inhibitor that induces apoptosis without generating an oxidative stress. In this case, Bcl 2 was again much more active than Hsp27. Fractionation studies indicated that, in L929 cells, Hsp27 is essentially present in the cytosol while Bcl-2 is membrane and mitochondria-associated. Hence, despite some similar cellular effects resulting from their expression, Bcl2 and Hsp27 polypeptides protect against oxidative stress and apoptosis with different efficiencies and by using different mechanisms. PMID- 11053667 TI - Age-related alterations of proteasome structure and function in aging epidermis. AB - Recent studies on the effect of aging in epidermal cells have evidenced a decrease of proteasome activity and content, suggesting that proteasome is down regulated in aged cells. The 20S proteasome is the major proteolytic system that has been implicated in removal of abnormal and oxidatively damaged proteins. Therefore, a decreased proteasome content may explain, at least in part, the well documented age-related accumulation of oxidized proteins. To gain further insight in other mechanisms that may be implicated in a decreased activity of the proteasome with age, 20S proteasome has been purified from the epidermis from donors of different ages: young, middle-aged and old. The patterns of proteasome subunits have been analyzed by 2D gel electrophoresis to determine whether its structure is also affected with age. The 2D gel pattern of proteasome subunits was found to be modified for four subunits, indicating that the observed decline in proteasome activity with age may also be related to alterations of its subunits. These subunit alterations are likely to be involved in the age-related decrease of proteasome activity since the specific peptidase activities of the purified proteasome were found to be decreased with age. PMID- 11053668 TI - Proteolysis of oxidised proteins and cellular senescence. AB - Aerobe living is consequently connected with a permanent oxidation of cellular proteins. Free radicals and other oxidants damage the normal intracellular protein pool. Therefore, the prevention of accumulation of oxidised cellular proteins is one of the major functions of the proteolytic machinery of mammalian cells. It is known that the multicatalytic proteinase complex, the proteasome, is the major protease that is able to recognise and degrade oxidised proteins. Cellular models are most useful to investigate biochemical changes of protein catabolism during senescence. Unfortunately, little is known about the protein turnover and the regulation of the proteasomal system as well as under oxidative stress conditions as during senescence. The proteasomal regulation during oxidative stress, protein oxidation and the changes of these processes during the ageing process are highlighted in this review. PMID- 11053669 TI - Modulating cellular aging in vitro: hormetic effects of repeated mild heat stress on protein oxidation and glycation. AB - Intracellular and extracellular proteins are subject to a variety of spontaneous non-enzymatic modifications which affect their structure, function and stability. Protein oxidation and glycation are tightly linked and are implicated in the development of many pathological consequences of aging. Although multiple endogenous pathways in the cell can prevent the formation of oxidized and glycated proteins, and repair and degrade abnormal proteins, such abnormal proteins do accumulate during aging. The heat shock response involving the family of stress-proteins or the so-called heat shock proteins (HSP), represents the quickest and highly conserved response to proteotoxic insults. Since repeated mild heat stress is able to prevent the onset of various age-related changes during cellular aging in vitro, we suggest that treatments which increase HSP expression should reduce the extent of accumulation of abnormal proteins during aging. Such modulation of aging is an example of hormesis, which is characterized by the beneficial effects resulting from the cellular responses to mild repeated stress. PMID- 11053670 TI - Does a retrograde response in human aging and longevity exist? AB - The retrograde response (RR) is a compensatory mechanism by which mutant strains of yeast are able to cope with mitochondrial DNA (mtDNA) impairments by up regulating the expression of the stress-responder nuclear genes and significantly increasing lifespan. Starting from the observation that both mtDNA variability and Tyrosine hydroxylase (THO, stress-responder gene) variability are correlated with human longevity, we asked ourselves whether mechanisms similar to RR may exist in humans. As a first investigative step we have analyzed the distribution of the mtDNA inherited variants (haplogroups) according to THO genotypes in three sample groups of increasing ages (20-49 years; 50-80 years; centenarians). We found that the mtDNA haplogroups and the THO genotypes are associated randomly in the first group, while in the second group, and particularly in the centenarians, a non-random association is observed between the mtDNA and nuclear DNA variability. Moreover, in centenarians the U haplogroup is over-represented (p=0.012) in subjects carrying the THO genotype unfavorable to longevity. On the whole these findings are in line with the hypothesis that longevity requires particular interactions between mtDNA and nuclear DNA and do not exclude the possibility that an RR has been maintained throughout evolution and it is present in higher organisms. PMID- 11053671 TI - Current status and perspectives of proteomics in aging research. AB - The accumulation of non-enzymatic modifications on both DNA and protein molecules under the attack of reactive oxygen species (ROS), is one of the most possible factors responsible for the functional deterioration in aged cells. Direct protein modifications as well as DNA damages may be detectable, in part, by proteome analysis if the gene expression is affected by the damages on DNA. The novel term "proteome", which is a compound of "protein" and "genome", means a whole set of proteins expressed in a tissue or a cell strain to be investigated. Proteomics is a methodology for analyzing proteomes. In proteomics, two dimensional gel electrophoresis is performed primarily to separate constitutive proteins, followed by mass spectrometry to identify each protein of interest and to determine a possible post-translational modification. Proteomics has offered us an innovative tool for investigating the molecular mechanisms of cellular aging. PMID- 11053672 TI - Uncoupling to survive? The role of mitochondrial inefficiency in ageing. AB - Mitochondria are incompletely coupled, and during oxidative phosphorylation some of the redox energy in substrates is lost as heat. Incomplete coupling is mostly due to a natural leak of protons across the mitochondrial inner membrane. In rat hepatocytes the futile cycle of proton pumping and proton leak is responsible for 20-25% of respiration; in perfused rat muscle the value is 35-50%. Mitochondrial proton cycling is estimated to cause 20-25% of basal metabolic rate in rats. Proton cycling is equally prominent in hepatocytes from several different mammalian and ectotherm species, so it may be a general pathway of ecologically significant energy loss in all aerobes. Because it occurs in ectotherms, thermogenesis cannot be its primary function. Instead, an attractive candidate for the function of the universal and expensive energy-dissipating proton cycle is to decrease the production of superoxide and other reactive oxygen species (ROS). This could be important in helping to minimise oxidative damage to DNA and in slowing ageing. Mitochondria are the major source of cellular ROS, and increased mitochondrial proton conductance leads to oxidation of ubiquinone and decreased ROS production in isolated mitochondria. However, to date there is no direct evidence in cells or organisms that mitochondrial proton cycling lowers ROS production or oxidative damage or that it increases lifespan. PMID- 11053673 TI - Heme oxygenase-1: role in brain aging and neurodegeneration. AB - The mechanisms responsible for excessive iron deposition and mitochondrial insufficiency in the aging and degenerating nervous system remain poorly understood. Heme oxygenase-1 (HO-1) is a 32kDa stress protein that degrades heme to biliverdin, free iron and carbon monoxide. Our laboratory has shown that cysteamine, dopamine, beta-amyloid, IL-1beta and TNF-alpha up-regulate HO-1 followed by mitochondrial sequestration of non-transferrin-derived 55Fe in cultured rat astroglia. In these cells and in rat astroglia transfected with the human HO-1 gene, mitochondrial iron trapping is abrogated by the HO-1 inhibitors, tin-mesoporphyrin and dexamethasone. We determined that HO-1 immunoreactivity is enhanced greatly in neurons and astrocytes of the hippocampus and cerebral cortex of Alzheimer subjects and co-localizes to senile plaques and neurofibrillary tangles (NFT). HO-1 staining is also augmented in astrocytes and decorates neuronal Lewy bodies in the Parkinson nigra. Collectively, our findings suggest that HO-1 over-expression contributes to the pathological iron deposition and mitochondrial damage documented in these aging-related neurodegenerative disorders. We recently observed that, paradoxically, HO-1 mRNA levels are markedly suppressed in peripheral lymphocytes of patients with early sporadic Alzheimer disease and may thus provide a useful biological marker of this condition. PMID- 11053674 TI - Modeling Alzheimer's disease in transgenic mice: effect of age and of presenilin1 on amyloid biochemistry and pathology in APP/London mice. AB - In transgenic mice that overexpress mutant Amyloid Precursor Protein [V717I], or APP/London (APP/Lo) (1999a. Early phenotypic changes in transgenic mice that overexpress different mutants of Amyloid Precursor Protein in brain. J. Biol. Chem. 274, 6483-6492; 1999b. Premature death in transgenic mice that overexpress mutant Amyloid precursor protein is preceded by severe neurodegeneration and apoptosis. Neuroscience 91, 819-830) the AD related phenotype of plaque and vascular amyloid pathology is late (12-15 months). This typical and diagnostic pathology is thereby dissociated in time from early symptoms (3-9 months) that include disturbed behavior, neophobia, aggression, glutamate excitotoxicity, defective cognition and decreased LTP. The APP/Lo transgenic mice are therefore a very interesting model to study early as well as late pathology, including the effect of age. In ageing APP*Lo mice, brain soluble and especially "insoluble" amyloid peptides dramatically increased, while normalized levels of secreted APPsalpha and APPsbeta, as well as cell-bound beta-C-stubs, remained remarkably constant, indicating normal alpha- and beta-secretase processing of APP. In double transgenic mice, i.e. APP/LoxPS1, clinical mutant PS1[A246E] but not wild type human PS1 increased Abeta, and plaques and vascular amyloid developed at age 6-9 months. The PS1 mutant caused increasing Abeta42 production, while ageing did not. Amyloid deposits are thus formed, not by overproduction of Abeta, but by lack of clearance and/or degradation in the brain of ageing APP/Lo transgenic mice. The clearance pathways of the cerebral amyloid peptides are therefore valuable targets for fundamental research and for therapeutic potential. Although hyper-phosphorylated protein tau was evident in swollen neurites around the amyloid plaques, neurofibrillary pathology is not observed and the "tangle" aspect of AD pathology is therefore still missing from all current transgenic "amyloid" models. Also the "ApoE4" risk for late onset AD remains a problem for modeling in transgenic mice. We have generated transgenic mice that overexpress human ApoE4 (2000. Expression of Human Apolipoprotein E4 in neurons causes hyperphosphorylation of Protein tau in the brains of transgenic mice. Am. J. Pathol. 156 (3) 951-964) or human protein tau (1999. Prominent axonopathy in the brain and spinal cord of transgenic mice overexpressing four-repeat human tau protein. Am. J. Pathol. 155, 2153-2165) in their neurons. Both develop a similar although not identical axonopathy, with progressive degeneration of nerves and with muscle wasting resulting in motoric problems. Remarkably, ApoE4 transgenic mice are, like the tau transgenic mice, characterized by progressive hyper phosphorylation of protein tau also in motor neurons which explains the motoric defects. Further crossing with the APP/Lo transgenic mice is ongoing to yield "multiple" transgenic mouse strains to study new aspects of amyloid and tau pathology. PMID- 11053675 TI - The processing and biological function of the human amyloid precursor protein (APP): lessons from different cellular models. AB - One of the major neuropathological hallmarks of Alzheimer's disease is the presence of senile plaques in vulnerable regions of CNS. These plaques are formed of aggregated amyloid peptide. Amyloid peptide is released by the cleavage of its precursor (APP). The establishment of cell lines expressing human APP allowed to characterize both amyloidogenic and non-amyloidogneic pathways of APP catabolism and to identify some of the proteins involved in this processing (known as secretases). This led to a better comprehension of amyloid peptide production, which needs to be further characterized since gamma-secretase is as yet not identified; moreover, we still lack a clear overview of the interactions between APP and other proteins promoting Alzheimer's disease (tau, presinilinsellipsis). An important limitation of these cell lines for studying the mechanisms involved in Alzheimer's disease is supported by the observation that human APP expression does not modify transfected cells survival. The infection of primary neuronal cultures with full-length human APP indicates that APP expression induces neuronal apoptosis by itself; this neurotoxicity does not rely on extracellular production of APP derivatives (secreted APP, amyloid peptide). It is now essential to understand, in neuronal models, the production, localization and involvement of amyloid peptide in neurodegenerative processes. PMID- 11053676 TI - Density profiles of Alzheimer disease regional brain pathology for the huddinge brain bank: pattern recognition emulates and expands upon Braak staging. AB - Density profiles of Alzheimer's disease (AD) regional brain pathology were constructed for 249 subjects in the Huddinge Brain Bank. Counts per square millimeter for neurofibrillary tangles (NFT), diffuse plaques (DP), and neuritic plaques (NP) in 38 areas were investigated using a pattern recognition technique called GoM. The seven distributional profiles of AD neuropathology emulated and expanded upon Braak staging illustrating induction (Groups 1-3) and clinical progression (Groups 4-7). Normal aging represented limited AD changes, few NFT in the entorhinal cortex and hippocampal CA1 (Group 1). The threshold for possible AD was NFT in the subiculum (Group 2), found with DP in the neocortex. Temporal medial NFT was the threshold for probable AD (Group 4). The 'oldest-old', often demented without brain atrophy, had extensive entorhinal/CA1 NFT and cortical DP, but few cortical NFT or NP (Group 5). A second subtype 'disconnection' (Group 6) lacked AD pathology for a specific set of subcortical and cortical areas. Accumulation of NFT in first-affected areas continued through end-stage disease (Group 7), with apparent rapid transition of DP to NP in the cortex during clinical progression. The evolution of AD is a highly ordered sequential process. Pattern recognition approaches such as GoM may be useful in better defining the process. PMID- 11053677 TI - Common gene variants, mortality and extreme longevity in humans. AB - Genetic factors influence variation in human life span. The fast technological advancements in genome research and the methodology for statistical analysis of complex traits provided new tools to unravel these genetic influences. Most of the genetic epidemiology and quantitative genetics is focused on the dissection of the genetic component of specific diseases rather than of human life span. Nevertheless, common variants of 22 genes have been tested for their contribution to mortality in the general population and extreme longevity in one or more studies. These studies provide indications as to the nature of biological pathways that might play a role in human ageing. Perhaps even more important at this time is the fact that they give valuable insights in the strengths and weaknesses of current strategies to identify gene variants affecting human life span and point at more powerful approaches. PMID- 11053678 TI - The network and the remodeling theories of aging: historical background and new perspectives. AB - Two general theories, i.e. "the network theory of aging" (1989) and "the remodeling theory of aging" (1995), as well as their implications, new developments, and perspectives are reviewed and discussed. Particular attention has been paid to illustrate: (i) how the network theory of aging fits with recent data on aging and longevity in unicellular organisms (yeast), multicellular organisms (worms), and mammals (mice and humans); (ii) the evolutionary and experimental basis of the remodeling theory of aging (immunological, genetic, and metabolic data in healthy centenarians, and studies on the evolution of the immune response, stress and inflammation) and its recent development (the concepts of "immunological space" and "inflamm-aging"); (iii) the profound relationship between these two theories and the data which suggest that aging and longevity are related, in a complex way, to the capability to cope with a variety of stressors. PMID- 11053679 TI - Congenital muscular dystrophy associated with calf hypertrophy, microcephaly and severe mental retardation in three Italian families: evidence for a novel CMD syndrome. AB - We describe four Italian patients (aged 3, 4, 12, and 13 years ) affected by a novel autosomal form of recessive congenital muscular dystrophy. These patients were from three non-consanguineous families and presented an almost identical phenotype. This was characterized by hypotonia at birth, joint contractures associated with severe psychomotor retardation, absent speech, inability to walk and almost no interest in their surroundings. In addition, all patients had a striking enlargement of the calf and quadriceps muscles. Ophthalmologic examination revealed no structural ocular abnormalities in any of the children; one patient had severe myopia. In all cases a magnetic resonance imaging of the brain showed an abnormal posterior cranial fossa with enlargement of the cisterna magna and variable hypoplasia of the vermis of the cerebellum. Abnormality of the white matter was also present in all patients, in the form of patchy signal most evident in the periventricular areas. Serum CK was grossly elevated in all. The muscle biopsy from all cases showed dystrophic changes compatible with congenital muscular dystrophy. Immunofluorescence studies showed mild to moderate partial deficiency of laminin alpha 2 chain. Linkage analysis in the only informative family excluded the known loci for congenital muscular dystrophy, including laminin alpha 2 chain on chromosome 6q2, the Fukuyama congenital muscular dystrophy locus on 9q3 and the muscle-eye-brain disease on chromosome 1p3. We propose that this represent a novel severe variant of congenital muscular dystrophy, with associated central nervous system involvement. PMID- 11053680 TI - Merosin-deficient congenital muscular dystrophy with mental retardation and cerebellar cysts unlinked to the LAMA2, FCMD and MEB loci. AB - We report a case of congenital muscular dystrophy with secondary merosin deficiency, structural involvement of the central nervous system and mental retardation in an 8-year-old girl from a consanguineous family. She had early onset hypotonia, generalized muscle wasting, with weakness especially of the neck muscles, joint contractures, mental retardation and high creatine kinase. Muscle biopsy showed dystrophic changes with partial deficiency of the laminin alpha(2) chain. Cranial magnetic resonance imaging revealed multiple small cysts in the cerebellum, without cerebral cortical dysplasia or white matter changes. The laminin alpha(2) chain (6q2), Fukuyama type congenital muscular dystrophy (9q31 q33) and muscle-eye-brain disease (1p32-p34) loci were all excluded by linkage analysis. We suggest that this case represents a new entity in the nosology of congenital muscular dystrophy. PMID- 11053681 TI - Secondary reduction in calpain 3 expression in patients with limb girdle muscular dystrophy type 2B and Miyoshi myopathy (primary dysferlinopathies). AB - Dysferlin is the protein product of the gene (DYSF) that is defective in patients with limb girdle muscular dystrophy type 2B and Miyoshi myopathy. Calpain 3 is the muscle-specific member of the calcium activated neutral protease family and primary mutations in the CAPN3 gene cause limb girdle muscular dystrophy type 2A. The functions of both proteins remain speculative. Here we report a secondary reduction in calpain 3 expression in eight out of 16 patients with a primary dysferlinopathy and clinical features characteristic of limb girdle muscular dystrophy type 2B or Miyoshi myopathy. Previously CAPN3 analysis had been undertaken in three of these patients and two showed seemingly innocuous missense mutations, changing calpain 3 amino acids to those present in the sequences of calpains 1 and 2. These results suggest that there may be an association between dysferlin and calpain 3, and further analysis of both genes may elucidate a novel functional interaction. In addition, an association was found between prominent expression of smaller forms of the 80 kDa fragment of laminin alpha 2 chain (merosin) and dysferlin-deficiency. PMID- 11053682 TI - Evaluation of heart involvement in gamma-sarcoglycanopathy (LGMD2C). A study of ten patients. AB - Thorough non-invasive cardiovascular studies were conducted in a series of ten gamma-sarcoglycanopathy Gypsy patients with the founder C283Y mutation in 13q12. Results were compared with those obtained in an age-matched group of normal boys and girls. The studies included electrocardiographic and echocardiographic evaluations using pulsed wave Doppler tissue imaging to assess regional diastolic function and myocardial velocities at various levels. This study confirms the significant electrocardiographic abnormalities described in previous studies. Furthermore, measurement of myocardial velocity at different levels demonstrated an abnormal relaxation pattern in the tricuspid annulus in four of the oldest patients, which strongly suggests intrinsic myocardial involvement of the right ventricle. To our knowledge, these specific studies have not been previously performed in a clinically and genetically homogeneous group of gamma sarcoglycanopathy patients and suggest primary myocardial involvement probably due to gamma-sarcoglycan deficiency in cardiac muscle fibres. Our results could be of interest in the follow-up of these patients and the prevention and treatment of late cardiological complications. PMID- 11053683 TI - Unusual expression of emerin in a patient with X-linked Emery-Dreifuss muscular dystrophy. AB - We report on a patient with the typical clinical findings of Emery-Dreifuss muscular dystrophy due to a mutation in the emerin gene that should have produced a higher molecular weight protein. Immunohistochemical analysis showed emerin localized only in the cytoplasm of muscle fibres and lymphoblastoid cells. The emerin molecule contained the nucleoplasmic domain and the transmembrane domain responsible for nuclear membrane targeting, so its incorrect localization and lack of function could be due to abnormal folding resulting in rapid degradation or inability to bind other nuclear proteins. PMID- 11053684 TI - Heterozygous myogenic factor 6 mutation associated with myopathy and severe course of Becker muscular dystrophy. AB - Myogenic factors (MYF) belong to the basic helix-loop-helix (bHLH) transcription factor family and regulate myogenesis and muscle regeneration. The physiological importance of both functions was demonstrated in homozygous Myf knockout mice and mdx mice. Myf5 and Myod are predominantly expressed in proliferating myoblasts while Myf4 and Myf6 are involved in differentiation of myotubes. In a boy with myopathy and an increase of muscle fibres with central nuclei we detected a heterozygous 387G-->T nucleotide transversion in the MYF6 gene (MIM*159991). Protein-protein interaction of mutant MYF6 was reduced, and DNA-binding potential and transactivation capacity were abolished, thus demonstrating MYF6 haploinsufficiency. The boy's father carried the identical mutation and, in addition, an in-frame deletion of exons 45-47 in his dystrophin gene. This mutation is normally associated with a mild to moderate course of Becker muscular dystrophy but the father suffered from a severe course of Becker muscular dystrophy suggesting MYF6 as a modifier. PMID- 11053685 TI - Hereditary motor and sensory neuropathy-Lom (HMSNL) in a Spanish family: clinical, electrophysiological, pathological and genetic studies. AB - The clinical, electrophysiological, pathological and genetic findings are described in the first Spanish family diagnosed with hereditary motor and sensory neuropathy type Lom (HMSNL) initially identified by Kalaydjeva et al. in 1996. The three affected patients belong to a non-consanguineous family with Gypsy background that were followed up over 10 years. Serial clinical and neurophysiological examinations and genetic analysis were undertaken in every patient. Sural nerve biopsy was performed in the oldest patient. The clinical features are similar to those previously described in HMSNL and all of them showed abnormal brain auditory evoked potentials. The oldest brother developed sensorineural deafness at the age of 20. Conduction velocities were unobtainable in all patients and nerves tested except for the median nerve in the youngest child in whom conduction was severely slowed. Neuropathological examination revealed a severely depleted nerve with very few surviving myelinated fibers which possessed thin myelin sheaths. Schwann cell processes were arranged in circular configurations without typical onion bulb configuration. Genetic analysis showed that the maternal chromosome inherited by all three affected siblings displayed a very unusual haplotype. Our patients show the characteristic clinical, electrophysiological and pathological findings described in HMSNL and represent the first reported Spanish family affected from the disease. The genetic findings in this family have contributed to refine the HMSNL critical linkage region. PMID- 11053686 TI - Hereditary motor and sensory neuropathy--Lom (HMSNL): refined genetic mapping in Romani (Gypsy) families from several European countries. AB - Hereditary motor and sensory neuropathy type Lom, initially identified in Roma (Gypsy) families from Bulgaria, has been mapped to 8q24. Further refined mapping of the region has been undertaken on DNA from patients diagnosed across Europe. The refined map consists of 25 microsatellite markers over approximately 3 cM. In this collaborative study we have identified a number of historical recombinations resulting from the spread of the hereditary motor and sensory neuropathy type Lom gene through Europe with the migration and isolation of Gypsy groups. Recombination mapping and the minimal region of homozygosity reduced the original 3 cM hereditary motor and sensory neuropathy type Lom region to a critical interval of about 200 kb. PMID- 11053687 TI - Charcot-Marie-Tooth 2-like presentation of an Algerian family with giant axonal neuropathy. AB - Giant axonal neuropathy is a rare autosomal recessive childhood disorder characterized by a peripheral neuropathy and features of central nervous system involvement. We describe four patients belonging to a consanguineous Algerian family with late onset (6-10 years) slowly progressive autosomal recessive giant axonal neuropathy. The propositus presented with a Charcot-Marie-Tooth 2-like phenotype with foot deformity, distal amyotrophy of lower limbs, areflexia and distal lower limb hypoesthesia. Central nervous system involvement occurred 10 years later with mild cerebellar dysarthria and nystagmus in the propositus and 16 years after onset, a spastic paraplegia in the oldest patient. The two youngest patients (13 and 8 years old) do not present any signs of central nervous involvement. Magnetic resonance imaging showed cerebellar atrophy in the two older. Nerve biopsy showed moderate axonal loss with several giant axons filled with neurofilaments. Genetic study established a linkage to chromosome 16q locus. This clinical presentation differs from the classical form of giant axonal neuropathy. PMID- 11053688 TI - Frequent mutations in Japanese patients with acid maltase deficiency. AB - We screened 22 Japanese patients with acid maltase deficiency (seven with the infantile type, eight with the juvenile type and seven with the adult type) for three previously described mutations, D645E, S529V and R672Q, and a novel mutation, R600C. Although D645E has been reported to be common in Chinese patients with the infantile type, only three of 44 alleles (two of 14 infantile type alleles) from Japanese patients harbored the D645E mutation. The S529V mutation was identified in six of 14 alleles from adult-onset patients. None of the infantile or juvenile patients harbored the S529V mutation. Therefore, S529V apparently results in the adult type disease and is common in Japanese adult onset patients. R672Q was identified in two pairs of siblings with the juvenile type. A novel mutation, R600C, was identified in eight of 22 patients (nine of 44 alleles). Therefore, R600C is another common Japanese mutation occurring at a CpG dinucleotide "hot spot". Homozygosity for this mutation apparently results in the infantile phenotype. Genetic diagnosis by detecting these four mutations might be feasible for most Japanese patients with acid maltase deficiency. PMID- 11053689 TI - Mitochondrial DNA variants in inclusion body myositis. AB - Mitochondrial DNA variants have been shown to be associated with many diseases. Mutations at mitochondrial DNA nucleotide positions 3192, 3196, 3397 and 4336 have been described in association with late-onset Alzheimer's disease. The pathological similarities between inclusion body myositis and Alzheimer's disease prompted an analysis of the relationship between the reported mutations and sporadic inclusion body myositis. The 4336G variant was not significantly increased in patients with inclusion body myositis or Alzheimer's disease when compared to controls. None of the patients with inclusion body myositis carried mutations at nucleotide positions 3192, 3196 and 3397. A transition at nucleotide position 4580 was detected in some patients with inclusion body myositis and Alzheimer's disease but was not significantly higher in frequency when compared to controls. Phylogenetic analysis showed that the 4336G and 4580A variants clustered together in their respective group. A group of patients with inclusion body myositis also clustered together on a separate branch of the phylogenetic tree. Closer investigation of this group revealed a common polymorphism at nucleotide position 16311. The frequency of the 16311C variant was higher in inclusion body myositis than in Alzheimer's disease and controls, although when only caucasian patients were considered the increased frequency was not statistically significant. Further studies will be required to determine whether this variant plays a role in the pathogenesis of inclusion body myositis. PMID- 11053690 TI - Altered pathological progression of diaphragm and quadriceps muscle in TNF deficient, dystrophin-deficient mice. AB - We have previously demonstrated a role for T cells in Duchenne muscular dystrophy (DMD) using the mdx mouse and have shown that T cell killing of dystrophic muscle can occur through perforin-dependent and perforin-independent mechanisms. In this investigation, we explore the possibility that one perforin-independent mechanism utilized by the T cells is cytokine-based killing, specifically by tumor necrosis factor (TNF). We tested this hypothesis by generating mice that are TNF-deficient and dystrophin-deficient (TNF-/mdx). Body mass and muscle mass of the TNF-/mdx mice were significantly less than TNF+/mdx mice at 8 weeks of age. Creatine kinase levels and overall muscle strength were unchanged. Histopathology measurements showed different results in the diaphragm and quadriceps muscles. These data suggest that removal of TNF in vivo in dystrophic mice has differential effects on diaphragm and quadriceps suggesting that TNF is an unfavorable target for immunotherapy for DMD. PMID- 11053691 TI - Relatedness, class, and social organization in a village in southern Thailand. AB - Darwinists still do not have a full theory of human social organization. Relatedness is a key idea in evolutionary theory. Yet, despite some pioneering studies, few Darwinists have used formal measures of relatedness, and even fewer have used them to describe social organization. Nobody has used relatedness to explain the rise, coherence, and decline of classes, class composition, and class relations. This article does that for one village in southern Thailand over a period of about 120 years. Each class had its own characteristic family type and relations between families, which varied over time. This article also uses the incidence of marriage between kin to show the need to take into account different perspectives on relatedness and social organization. PMID- 11053692 TI - Scientific discoveries as cultural displays: a further test of Miller's courtship model. AB - The biographies of 280 scientists demonstrate that the age distribution of their career peaks is identical to those of jazz musicians, painters, and writers, and that this universal age profile holds only among scientists who were married some time in their lives. I interpret this as support for Miller's cultural display model in an evolutionarily novel environment. PMID- 11053693 TI - Number of children desired and preferred spousal age difference: context-specific mate preference patterns across 37 cultures. AB - Men universally express a preference for youth in a long-term mate, presumably an evolved desire originating from the close and recurrent statistical association between a woman's age and her residual reproductive value (future reproductive potential). As a consequence, we hypothesized a positive correlation for men (but not women) between the number of children desired and preferred spousal age difference - a context-specific shift in mate preference depending on whether the man is pursuing a "quality" or "quantity" reproductive strategy. We tested this hypothesis with data provided by 9809 participants from 37 cultures located in six continents and five islands. Between-culture analyses confirmed the hypothesis, even after statistically controlling for preferred age at first marriage, current age of participant, and current marital status. Discussion notes limitations and focuses on other possible context-sensitive shifts in mate preferences. PMID- 11053694 TI - The ratio of 2nd to 4th digit length and male homosexuality. AB - Sexual orientation may be influenced by prenatal levels of testosterone and oestrogen. There is evidence that the ratio of the length of 2nd and 4th digits (2D:4D) is negatively related to prenatal testosterone and positively to oestrogen. We report that (a) 2D:4D was lower in a sample of 88 homosexual men than in 88 sex- and age-matched controls recruited without regard to sexual orientation, (b) within the homosexual sample, there was a significant positive relationship between mean 2D:4D ratio and exclusive homosexuality, (c) overall, there was a decrease in 2D:4D from controls to homosexual men to bisexual men and (d) fraternal birth order, a positive predictor of male homosexuality, was not associated with 2D:4D in a sample of 240 Caucasian men recruited without regard to sexual orientation and 45 homosexual men.Further work is needed to confirm the relationships between 2D:4D and sexual orientation. However, these and other recent data tend to support an association between male homosexuality and high fetal testosterone. Very high testosterone levels may be associated with a sexual preference for both men and women. PMID- 11053695 TI - Diabetes mellitus, impaired fasting glucose, atherosclerotic risk factors, and prevalence of coronary heart disease. AB - Patients with diabetes mellitus (DM), both diagnosed (history of) and undiagnosed (by fasting glucose [FG] only), as well as impaired FG have an increased risk of coronary heart disease (CHD), compared with those with normal FG. Elevations in FG levels, even in normoglycemic subjects (<110 mg/dl), may be significantly related to CHD morbidity and mortality. Improving lipid profiles and blood pressure can decrease both CHD morbidity and mortality in these patients. We evaluated the relation of glucose status to lipid levels, other risk factors, and prevalence of CHD using the 1997 American Diabetes Association diagnostic criteria in a representative sample of United States adults studied in the Third National Health and Nutrition Examination Survey from 1988 to 1994. Impaired FG, diagnosed DM, and undiagnosed DM were more prevalent in older age groups; those > or =65 years had increased prevalence compared with those <50 years old (rate ratios for IFG, DM-FG, and history of DM were 3.5, 4.8, and 10.8, respectively). Glycosylated hemoglobin levels were increased by glucose status. The frequency of known CHD risk factors also increased with worsening glucose status. Age-adjusted CHD prevalence was increased with impaired FG (rate ratio 1.47), DM-FG (rate ratio 1.56), and history of DM (rate ratio 1.72), compared with normal FG. Adjusting for age and other CHD risk factors, hyperglycemia was no longer significantly associated with CHD prevalence. Lipid values, especially high density lipoprotein cholesterol, hypertension, and other CHD risk factors were more strongly associated with CHD than glucose status. Thus, patients with impaired FG, DM-FG, and history of DM should be considered at higher risk for CHD morbidity and mortality. However, hyperglycemia, per se, does not explain the excess risk. In addition to glucose, lipid profiles and blood pressure should be periodically monitored and appropriate treatment provided to reduce morbidity and mortality from CHD. PMID- 11053696 TI - Prognostic significance of mild mitral regurgitation by color Doppler echocardiography in acute myocardial infarction. AB - Mitral regurgitation (MR) complicating acute myocardial infarction (AMI) is associated with increased mortality. The prognostic significance of only mild MR detected by echocardiography in patients with AMI is unknown. This study assessed the long-term risk associated with mild MR detected by color Doppler echocardiography within the first 48 hours of admission in 417 consecutive patients with AMI. No MR was detected in 271 patients (65%), mild MR was seen in 121 patients (29%), and moderate or severe MR was noted in 25 patients (6%). One year mortality rates were 4.8%, 12.4%, and 24%, respectively (p<0.001). Multivariate analysis revealed that mild MR was independently associated with increased 1-year mortality (p<0.05) after adjustment for age, gender, previous myocardial infarction, diabetes mellitus, systemic hypertension, Killip grade > or =2 on admission, and left ventricular ejection fraction < or =40%. The hazard ratio for 1-year mortality was 2.31 (95% confidence interval 1.03 to 5.20) for mild MR and 2.85 (95% confidence interval 0.95 to 8.51) for moderate or severe MR. Thus, mild MR detected by color Doppler echocardiography within the first 2 days of admission in patients with AMI is a significant independent risk predictor for 1-year all-cause mortality. PMID- 11053697 TI - Clinical trials of unfractionated heparin and low-molecular-weight heparin in addition to aspirin for the treatment of unstable angina pectoris: do the results apply to all patients? AB - We sought to determine how the results of available randomized controlled trials of intravenous unfractionated heparin (UH) and low-molecular weight heparin (LMWH) apply to unselected patients with unstable angina pectoris (UAP). Although UH is widely used in addition to aspirin for treatment of UAP, the evidence is weak for a net benefit over aspirin alone. LMWH preparations may confer a net benefit over UH for the treatment of UAP in clinical trials. It is not clear, however, how trial results are generalized to unselected patients with UAP. Using criteria from the Agency for Health Care Policy and Research Unstable Angina Clinical Practice Guideline, we identified 277 consecutive patients with primary UAP. Exclusion criteria were applied from 6 trials of UH in addition to aspirin and 5 trials of LMWH in addition to aspirin for the treatment of UAP. Clinical outcomes were compared among ineligible and eligible patients for trial enrollment. Patients meeting exclusion criteria were older and had more extensive coexisting medical illness than eligible patients for trial enrollment. Thirty eight percent to 42% of our study population met > or = 1 exclusion criteria for each of the 6 trials of UH, and 14% to 46% met > or = 1 exclusion criteria for each of the 5 LMWH trials. The 1-year all-cause death rate was higher in UH ineligible patients compared with UH eligible patients (16% vs 4%, p = 0.003) and in LMWH ineligible patients compared with LMWH eligible patients (16% vs 7%, p = 0.005). Thus, clinical trials of UH and LMWH may have limited generalizability to unselected patients with UAP, many of whom have characteristics that would exclude them from trial enrollment and put them at risk for adverse outcomes. PMID- 11053698 TI - Sites of interleukin-6 release in patients with acute coronary syndromes and in patients with congestive heart failure. AB - This study examines the source of elevated interleukin-6 (IL-6) levels in patients with acute coronary syndrome (ACS) and congestive heart failure (CHF). IL-6 is elevated in the peripheral blood of patients with ACS and CHF, but it is not known if this proinflammatory cytokine is from a cardiac or extracardiac source. Blood samples were obtained from the femoral artery, femoral vein, left main coronary artery, and coronary sinus in 57 patients during cardiac catheterization. IL-6 levels from 12 patients with ACS and 12 patients with CHF were compared with the IL-6 levels in 33 patients who had neither of these clinical conditions. Median IL-6 levels in the peripheral and coronary circulation were a minimum fivefold higher in patients with ACS or CHF relative to control patients. An elevated transcardiac IL-6 gradient (coronary sinus-left main level) was present in patients with ACS (median 5.2; 25th and 75th percentiles 3.9 and 29.3 pg/ml, respectively) compared with control patients (median 0, -0.7 and 0.5 pg/ml; p < 0.001), but not in patients with CHF (median 0.4, -0.7 and 3.5 pg/ml; p = NS). Elevated IL-6 levels in patients with ACS derive from a cardiac source, presumably from "inflamed" coronary plaques and areas of myocardial necrosis, whereas elevated levels in patients with CHF are most likely the result of extracardiac production. PMID- 11053699 TI - Transthoracic Doppler echocardiographic comparison of left internal mammary grafts to left anterior descending coronary artery with ungrafted right internal mammary arteries in patients with and without myocardial ischemia by dobutamine stress echocardiography. AB - To characterize Doppler flow patterns of the grafted left internal mammary artery (LIMA) in patients with and without dobutamine stress induced wall motion abnormalities in the graft distribution, we studied 29 patients who underwent coronary artery bypass surgery using LIMA grafts to the left anterior descending coronary artery (LAD). The ungrafted right internal mammary artery (RIMA) was used as a control. RIMA Doppler flow pattern was predominantly systolic in all patients. In patients without ischemia in the LAD distribution, LIMA flow was predominantly diastolic. In patients with ischemia, LIMA flow was predominantly systolic. In the grafted LIMA, a ratio of diastolic to systolic time-velocity integral of > 1.5 best showed absence of ischemia in the graft distribution. In summary, characterization of the Doppler flow pattern in the internal mammary arteries is feasible. In the grafted LIMA, ratios of diastolic to systolic flow are less in patients with an ischemic response in the subtended vascular bed than in those without ischemia. PMID- 11053700 TI - Assessing a strategy of initial stand-alone extractional atherectomy followed by staged stent placement in degenerated saphenous vein graft lesions. AB - To assess whether a staged strategy (initial stand alone transluminal extraction atherectomy and coumadin therapy followed by stenting six weeks later) could reduce ischemic complications in degenerated saphenous vein graft (SVG) interventions, we studied 72 patients undergoing percutaneous interventions of degenerated SVG. Patients were divided into two groups; 28 were treated with a staged strategy (group I) and 44 with similar lesion characteristics were treated with a definitive initial procedure with transluminal extraction atherectomy +/- adjunctive balloon angioplasty and stenting (group II). Procedural success, major in-hospital complications (death, Q-wave myocardial infarction, and emergent coronary bypass surgery), and incidence of distal embolization were compared between the 2 groups. Procedural success was lower (92% vs 100%, p = 0.14) and major in-hospital complications were higher (0% vs 11%, p = 0.14) in group II. Distal embolization occurred in 11% of the patients in group I compared with 23% of the patients in group II (p = 0.19). At 6 week follow-up (group I), 9 patients (33%) had negative symptoms, 11 (41%) underwent stent implantation, 3 (11%) did not require any further therapy (without significant stenosis), and 4 (14%) had total occlusions. We therefore conclude that this staged strategy in degenerated SVG appears to reduce distal embolization but most importantly avoids major in hospital complications, including any deaths either at the time of initial procedure or during the 6-week follow-up period. PMID- 11053701 TI - Long-term follow-up after coronary stenting and intravascular red laser therapy. AB - A high restenosis rate remains a limiting factor for coronary angioplasty and stenting. Recently, use of intravascular red light therapy (IRLT) has been shown to be effective in different animal models and in humans in reducing the restenosis rate. Sixty-eight patients were treated with IRLT in conjunction with coronary stenting procedures. Mean age was 64 +/- 9 years. Treated lesions were type A (11), type B (42), and type C (18) with a mean lesion length of 16.5 +/- 2.4 mm. Reference vessel diameter and minimal lumen diameter (MLD) before therapy were 2.90 +/- 0.15 and 1.12 +/- 0.36 mm, respectively. After stenting and laser irradiation, MLD was 2.76 +/- 0.39 mm. No procedural complications or in-hospital adverse events occurred. All patients were followed up as depicted in the protocol. Sixty-one patients underwent angiographic restudy, which revealed restenosis in 9 patients (14.7%). Observed restenosis rate by artery size was > 3 mm (n = 21, 0%), 2.5 to 3.0 mm (n = 28, 14.2%), and <2.5 mm (n = 12, 41.6%). We conclude that IRLT is safe and feasible and reduces the expected restenosis rate in patients after coronary stenting in arteries of >2.5 mm. PMID- 11053702 TI - Comparison of percutaneous interventions for unstable angina pectoris in patients with and without previous coronary artery bypass grafting. AB - An increasing number of patients who have undergone previous coronary artery bypass grafting (CABG) are referred for percutaneous coronary revascularization. We identified patients who underwent percutaneous intervention for unstable angina from 1990 to 1998 at our institution and assigned them into 2 groups based on whether or not they had undergone previous CABG. There were 1,431 patients with and 4,629 patients without previous CABG. Previous CABG patients were older, had more atherosclerotic risk factors, more heart failure, lower ejection fraction, more multivessel disease, more multilesion treatment, more complex lesions, and less complete revascularization. Adjusting for baseline differences, previous CABG was associated with worse long-term mortality (RR 1.47, 95% confidence intervals [CI] 1.22 to 1.77, p < 0.001) and death, myocardial infarction, and/or revascularization (RR 1.16, 95% CI 1.04 to 1.30, p = 0.01); treatment of native lesions in patients with previous CABG versus treatment of vein graft lesions was associated with a reduction in this composite end point (RR 0.75, 95% CI 0.65 to 0.87, p < 0.001). Post-CABG patients treated between 1995 and 1998 had lower long-term mortality (RR 0.76, 95% CI 0.59 to 0.99, p = 0.04) and death, myocardial infarction, and/or revascularization (RR 0.76, 95% CI 0.66 to 0.88, p < 0.001) compared with those treated between 1990 and 1994. Thus, in patients with unstable angina referred for percutaneous revascularization, previous CABG is associated with reduced event-free survival, although the outcome of post-CABG patients treated from 1995 to 1998 is superior to that observed in patients treated from 1990 to 1994. In patients who underwent previous CABG, treatment of native lesions affords better long-term outcome than vein graft intervention. PMID- 11053703 TI - Relation between coronary artery disease, baseline clinical variables, revascularization mode, and mortality. CABRI Participants. Coronary Angioplasty vs. Bypass Revascularisation Investigation. AB - The Coronary Angioplasty vs. Bypass Revascularisation Investigation (CABRI) trial comparing percutaneous transluminal coronary angioplasty (PTCA) with coronary artery bypass grafting did not show a difference in mortality with either procedure. Nonrandomized studies suggest that coronary artery disease (CAD) severity and distribution influences outcome. In the present study we explored the effect of prerevascularization CAD on 1-year mortality in the CABRI population, while adjusting for other baseline variables. Of the 1,054 patients recruited, there were sufficient angiographic results to derive the CAD scores in 974 (92.4%). Of these 974, there were 32 deaths. A number of CAD scores, both weighted for proximal disease (Duke and Leaman) and nonweighted, were used. These scores were then cross-tabulated against mortality. Demographic and clinical variables were also cross-tabulated against mortality and used to derive an initial logistic regression model to predict mortality. The effect of adding each of the CAD scores to this initial model was then assessed. After inclusion of the CAD scores, the best model was: (1) presence of peripheral vascular disease (odds ratio [OR] 3.89, p = 0.0025), (2) previous cerebrovascular accident (OR 2.86, p = 0.043), (3) older age (OR 1.05, p = 0.039), (4) a higher Duke score (OR 2.84, p = 0.0061), and (5) having undergone PTCA (OR 2.12, p = 0.047). In the CABRI population, adjustment for baseline variables, including prerevascularization CAD, revealed significantly higher mortality in those who underwent PTCA than in those who underwent coronary artery bypass grafting. PMID- 11053704 TI - Evidence that triglycerides are an independent coronary heart disease risk factor. AB - In the past, the relation between hypertriglyceridemia and coronary heart disease (CHD) has been uncertain. However, a recent multivariate analysis of 8-year follow-up data from the large-scale Prospective Cardiovascular Munster study found hypertriglyceridemia to be an independent risk factor for major coronary events after controlling for low-density lipoprotein (LDL) and high-density lipoprotein (HDL) cholesterol. Hypertriglyceridemia combined with elevated LDL cholesterol and high LDL:HDL cholesterol ratio (>5) increased the CHD event risk by approximately sixfold. Similarly, a large meta-analysis of 17 prospective trials reported hypertriglyceridemia to be an independent risk factor for cardiovascular disease. In this study, an 88 mg/dl (1.0 mmol/L) increase in plasma triglyceride levels significantly increased the relative risk of cardiovascular disease by approximately 30% in men and 75% in women; the corresponding rates were somewhat lower (14% and 37%) but still statistically significant after adjustment for HDL cholesterol level. These data and observations from patients in the Helsinki Heart Study and the Stockholm Ischemic Heart study, that the greatest coronary benefit during lipid-lowering drug therapy occurred among hypertriglyceridemic patients, argue strongly for an independent role for hypertriglyceridemia in CHD risk. In the recent Veterans Affairs Cooperative Studies Program High-Density Lipoprotein Cholesterol Intervention Trial, the use of gemfibrozil to raise HDL cholesterol levels and lower levels of triglycerides without lowering LDL cholesterol levels reduced coronary events in men with established CHD, whereas preliminary results from the Bezafibrate Infarction Prevention Trial indicate a reduction in coronary end points in patients with elevated baseline triglyceride levels. To achieve the greatest possible reduction in CHD risk, antihyperlipidemic treatment strategies should also be aimed at reducing elevated triglycerides. PMID- 11053705 TI - Comparison of intravenous flecainide, propafenone, and amiodarone for conversion of acute atrial fibrillation to sinus rhythm. AB - In a prospective, single-blind trial, we randomized 150 consecutive symptomatic patients with acute (< or = 48 hours' duration) atrial fibrillation to receive intravenous flecainide, propafenone, or amiodarone. Flecainide and propafenone were administered as a bolus dose of 2 mg/kg in 20 minutes. A second bolus dose of 1 mg/kg in 20 minutes was administered if conversion to sinus rhythm was not achieved after 8 hours. Amiodarone was administered as a bolus of 5 mg/kg in 20 minutes followed by a continuous infusion of 50 mg/hour. By the end of a 12-hour observation period, conversion to sinus rhythm was achieved in 45 patients (90%) in the flecainide group, 36 (72%) in the propafenone group, and 32 (64%) in the amiodarone group (p = 0.008 for the overall comparison, p = 0.002 for flecainide vs amiodarone, p = 0.022 for flecainide vs propafenone, and p = 0.39 for propafenone vs amiodarone). When compared with amiodarone, this higher reversion rate with flecainide was present from the first hour of the study period. However, only after administering the second bolus was there a significant difference between flecainide and propafenone. Median time to conversion to sinus rhythm was different among groups (p < 0.001), and it was lower in the flecainide (25 minutes; range 4 to 660) and propafenone (30 minutes; range 10 to 660) groups than in the amiodarone group (333 minutes; range 15 to 710; p < 0.001 for both comparisons). Flecainide, at the doses administered in this study, is more effective than propafenone and amiodarone for conversion of acute atrial fibrillation to sinus rhythm. Propafenone and amiodarone have similar conversion rates, although propafenone was faster in achieving the conversion to sinus rhythm. PMID- 11053706 TI - Relation of autonomic modulation to recurrence of atrial fibrillation following cardioversion. AB - The aim of this study was to investigate the time course of changes in autonomic nervous system activity in patients with long-standing atrial fibrillation (AF) following internal electrical conversion to sinus rhythm and to look for differences between patients who do and do not relapse. Time-domain indexes of heart rate variability were calculated from 24-hour Holter recordings on the day of conversion and 1 day and 1 month afterward for 22 patients with chronic (> 3 months) AF. Ten healthy subjects served as a control group. During the day of cardioversion the mean RR interval and its circadian variation differed significantly between controls and patients. The mean values of successive RR intervals that deviated by > 50% from the prior RR interval and the root-mean square of successive RR interval differences--indexes of vagal modulation--were initially significantly higher in patients than in controls but showed a decrease (p < 0.05) by the second day (from 12.4 +/- 7% to 8.1 +/- 5% to 7.3 +/- 5% and from 49 +/- 9 to 39 +/- 12 to 41 +/- 11 ms, respectively) to levels similar to those of the controls (7.6 +/- 5% and 40 +/- 17 ms, respectively). Only these 2 indexes contained significant prognostic information about relapse: patients who later relapsed had higher initial values than those who did not, and these values remained high during the 2 days after conversion. In conclusion, this study provides data confirming that spontaneous chronic AF in humans results in a significant increase in vagal tone that is reversed with time after restoration of sinus rhythm. Persistently higher values of vagal tone are observed in patients who relapse, and are probably a predictor for recurrence. PMID- 11053707 TI - Heart rate variability patterns before ventricular tachycardia onset in patients with an implantable cardioverter defibrillator. Participating Investigators of ICD-HRV Italian Study Group. AB - Time- and frequency-domain analysis of heart rate variability (HRV) has been proven effective in describing alteration of autonomic control mechanisms and in identifying patients with increased cardiac and arrhythmic mortality. Patients with implantable cardioverter defibrillators offer the opportunity to evaluate HRV patterns before ventricular tachycardia (VT) and under control conditions. We therefore analyzed time- and frequency-domain parameters of short-term HRV and power-law behavior of RR interval time series at rest, at 15 to 30 minutes, and immediately before VT. In comparison to control conditions, lower values of mean cycle length duration and total power were observed before VT. Spectral analysis indicated that the low- to high-frequency ratio was significantly higher (5.5 +/- 0.6 vs 2.8 +/- 0.3) immediately before VT than during rest. Both findings were consistent with the shift of sympathovagal balance toward sympathetic predominance and reduced vagal tone. Before VT, a more negative value of the scaling exponent beta of the power-frequency relation (-1.57 +/- 0.04 vs -1.33 +/ 0.04) also confirmed the presence of an altered HRV pattern in comparison to controls. Thus, both abnormal autonomic modulation and dynamic patterns of HRV seem to characterize the minutes before arrhythmia onset in these patients. PMID- 11053708 TI - Two-dimensional echocardiographic assessment of right ventricular function as a predictor of outcome in hypoplastic left heart syndrome. AB - This study was undertaken to assess the importance of right ventricular function at the time of initial presentation on early and intermediate outcome in patients with hypoplastic left heart syndrome (HLHS). Several studies have attempted to define physiologic risk factors for poor early outcome following the Norwood palliation for HLHS. No clinical or hemodynamic factors including right ventricular function have been found to reliably predict Norwood I operative survival. The relation between initial ventricular function and later survival has not been investigated. To assess the importance of right ventricular (RV) function at the time of initial presentation on outcome in patients with HLHS, systolic function was determined by qualitative and quantitative methods in 60 consecutive patients before surgical intervention. The effects on stage I operative survival, survival to stage II, and overall survival were analyzed. Initial RV function did not impact on stage I survival. However, analysis of later outcome of the stage I survivors showed that those with prestage I RV dysfunction had significantly greater mortality before stage II. Actuarial survival 18 months after Norwood surgery was 93% for patients with initially normal RV function compared with 47% for those with abnormal function (p = <0.005). The relative risk for later mortality was approximately 11 times greater for patients with initial RV dysfunction. Thus, RV dysfunction identifiable soon after initial presentation does not impact on early survival after Norwood I operation for HLHS. Intermediate and overall survival, however, is significantly decreased in patients with initially diminished RV function. PMID- 11053709 TI - Location of acutely successful radiofrequency catheter ablation of intraatrial reentrant tachycardia in patients with congenital heart disease. AB - Intraatrial reentrant tachycardia (IART) is common after surgery for congenital heart disease (CHD). Radiofrequency (RF) catheter ablation of IART targets anatomic areas critical to the maintenance of the arrhythmia circuit, areas that have not been well defined in this patient population. The purpose of this study was to determine the anatomic areas critical to IART circuits, defined by activation mapping and confirmed by an acutely successful RF ablation at the site. A total of 110 RF ablation procedures in 88 patients (median age 23.4 years, range 0.1 to 62.7) with CHD were reviewed. Patients were grouped according to surgical intervention: Mustard/Senning (n = 15), other biventricular repaired CHD (n = 24), Fontan (n = 43), and palliated CHD (n = 6). In first-time ablation procedures, > or = 1 IART circuits were acutely terminated in 80% of Mustard/Senning, 71% of repaired CHD, and 72% of Fontan (p = NS). The palliated CHD group underwent 1 of 6 successful procedures (17%), and this patient was excluded. The locations of acutely successful RF applications in Mustard/Senning patients (n = 14 sites) were at the tricuspid valve isthmus (57%) and at the lateral right atrial wall (43%). In patients with repaired CHD (n = 18 sites), successful RF sites were at the isthmus (67%) and the lateral (22%) and anterior (11%) right atria. In the Fontan group (n = 40 sites), successful RF sites included the lateral right atrial wall (53%), the anterior right atrium (25%), the isthmus area (15%), and the atrial septum (7%). Location of success was statistically different for the Fontan group (p = .002). In conclusion, the tricuspid valve isthmus is a critical area for ablation of IART during the Mustard/ Senning procedure and in patients with repaired CHD. IART circuits in Fontan patients are anatomically distinct, with the lateral right atrial wall being the more common area for successful RF applications. This information may guide RF and/or surgical ablation procedures in patients with CHD and IART. PMID- 11053710 TI - Relation of regional sympathetic denervation and myocardial perfusion disturbance to wall motion impairment in Chagas' cardiomyopathy. AB - Impairment of sinus node autonomic control and myocardial perfusion disturbances have been described in patients with chronic Chagas' cardiomyopathy. However, it is not clear how these conditions contribute to myocardial damage. In this investigation, iodine-123 (I-123) meta-iodobenzylguanidine (MIBG) and thallium 201 myocardium segmental uptake were studied in correlation with the severity of left ventricular (LV) dysfunction detected in various phases of Chagas' heart disease. Group I consisted of 12 subjects (43 +/- 4 years, 7 men) with no symptoms and no cardiac involvement on electrocardiogram (ECG) or echocardiography; group II consisted of 13 patients (48 +/- 3 years, 9 men) with abnormal resting ECG and/or echocardiographic segmental abnormalities, and LV ejection fraction of > or = 0.5; group III was comprised of 12 patients (59 +/- 3 years, 10 men) with more severe heart disease, LV dilation, and LV ejection fraction of < 0.5. Eighteen control volunteers (38 +/- 3 years, 9 men) were also included in the study. I-123 MIBG single-photon emission computed tomographic (SPECT) segmental uptake defects were observed in group I (33%), group II (77%), and group III (92%). Quantitative analysis showed mean areas of reduced LV I-123 MIBG uptake: group I was 3.7 +/- 2.1%; group II was 8.3 +/- 2.3%; and group III was 19.0 +/- 3.3%. The differences between group I and both groups II and III were statistically significant (p < 0.001, analysis of variance test). Myocardial perfusion defects (reversible, fixed, and paradox) were observed in group I (83%), group II (69%), and group III (83%). A marked topographic association between perfusion, innervation, and wall motion abnormalities (assessed by gated SPECT perfusion studies) was observed in all the groups. Defects predominated in the inferior, posterior lateral, and apical LV regions. Thus, extensive impairment of cardiac sympathetic function at the ventricular level occured early in the course of Chagas' cardiomyopathy and was related to regional myocardial perfusion disturbances, before wall motion abnormalities. Both conditions are associated with progression of ventricular dysfunction. PMID- 11053711 TI - Hemodynamic performance during maximum exercise in adult patients with the ross operation and comparison with normal controls and patients with aortic bioprostheses. AB - This study examines the resting and exercise hemodynamic performance of the pulmonary autografts in the aortic position as well as of the homografts used for right ventricular outflow reconstruction in patients undergoing the Ross operation. Previous studies have reported excellent resting hemodynamics in patients who underwent aortic valve replacement with a pulmonary autograft. However, there are very few studies of their hemodynamic performance during exercise. Twenty adult subjects who underwent the Ross operation and 12 normal control subjects were submitted to maximum romp bicycle exercise. The valve effective orifice areas and transvalvular gradients of both aortic (autograft) and pulmonary (homograft) valves were measured at rest and at peak of maximum exercise using Doppler echocardiography. Valve areas were indexed for body surface area. The hemodynamics of the aortic valve were very similar in Ross subjects and in control subjects at rest and during exercise. However, the indexed valve area of the pulmonary valve at rest was significantly (p < 0.001) lower in the Ross subjects (1.10 +/- 0.46 cm2/ m2) than in the control subjects (1.95 +/- 0.41 cm2/m2), resulting in higher (p = 0.004) mean gradients at rest (Ross: 9 +/- 7 mm Hg vs control: 2 +/- 1 mm Hg) and at peak exercise (Ross: 21 +/ 14 mm Hg vs control: 7 +/- 2 mm Hg). The pulmonary autograft provided excellent hemodynamics in the aortic position either at rest or during maximum exercise, whereas moderately high gradients were found during exercise across the homograft implanted in the pulmonary valve position. Future improvement of the Ross procedure should be oriented toward the search of new methods to prevent the deterioration of the homografts. PMID- 11053712 TI - Harvey Stanley Hecht, MD: a conversation with the editor. PMID- 11053713 TI - Comparison of in-hospital outcomes after coronary angioplasty with or without stent placement for acute myocardial infarction. AB - This study compared the in-hospital outcomes of patients treated with or without stent placement during mechanical revascularization for acute myocardial infarction. After correction for differences in baseline characteristics, patients treated with stent placement had lower in-hospital mortality. PMID- 11053714 TI - Meta-analysis of the association of platelet glycoprotein IIIa PlA1/A2 polymorphism with myocardial infarction. AB - This meta-analysis examined all the published reports up to October 1999 that studied the association between PlA2 polymorphism of platelet glycoprotein IIIa gene and myocardial infarction. The PlA2 polymorphism was not found to be associated with an increased risk of myocardial infarction, either overall or in selected subgroups, which were patients with premature disease onset (age < or = 60 years), first acute myocardial infarction, and patients who were men, women, and exclusively Caucasian. PMID- 11053715 TI - Homocysteine, a risk factor for coronary artery disease or not? A meta-analysis. AB - This meta-analysis suggests that homocysteine may not be as harmful for the heart as it seems. At the same time, however, homocysteine may be an indicator for unhealthy lifestyles, and therefore, an important variable for cardiologists to take into account when assessing coronary artery disease. PMID- 11053716 TI - Comparison of modes of death and cardiac necropsy findings in fatal acute myocardial infarction in men and women >75 years of age. AB - In comparing the cause of death and other cardiac morphologic findings among 60 women and 40 men aged >75 years who died of acute myocardial infarction, we found that women died more often from mechanical complications than left ventricular pump failure. Women had cardiomegaly, nonanterior location of acute myocardial infarction, healed myocardial infarcts, and dilated left ventricular cavity less often than men. PMID- 11053717 TI - Use of fractional myocardial flow reserve to assess the functional significance of intermediate coronary stenoses. AB - The goal of the present study was to compare the use of pressure-derived myocardial fractional flow reserve for detecting ischemia with nuclear stress imaging in patients undergoing stent placement for intermediate coronary lesions. We demonstrated that myocardial fractional flow reserve detects ischemia in intermediate coronary lesions accurately when compared with nuclear stress imaging. PMID- 11053718 TI - Serial intravascular ultrasound and quantitative coronary angiography after self expandable wallstent coronary artery implantation. AB - Serial intracoronary ultrasound studies revealed significant postprocedural stent expansion accompanied by significant stent shortening during long-term follow-up. The disadvantageous lumen loss by neointimal formation could be balanced by late stent expansion. PMID- 11053719 TI - Causes of early reintervention after successful coronary artery stenting. AB - Acute reintervention was performed in 26 of 1,620 patients after coronary stenting (1.6%). Half of the patients had stent thrombosis and the other half residual anatomic problems. The mean time for reintervention was shorter in patients with stent thrombosis. All patients with stent thrombosis had a sudden recurrence of chest pain. Electrocardiographic changes were more common with stent thrombosis. Composite end point occurred in 10 patients (77%) with stent thrombosis versus 5 (39%) in the other group (p = 0.04). PMID- 11053720 TI - Effect of off-pump coronary artery bypass grafting on morbidity. AB - Three hundred three patients who underwent off-pump coronary bypass were compared with 483 patients who underwent standard on-pump coronary bypass. There was a significant reduction in intraoperative blood transfusion requirements, as well as a significant reduction in the incidence of neurologic, renal, and prolonged ventilatory complications in the off-pump group. PMID- 11053721 TI - Time course of recovery of left atrial mechanical dysfunction after cardioversion of spontaneous atrial fibrillation with the implantable atrial defibrillator. AB - The effect of the timing of cardioversion of atrial fibrillation on left atrial mechanical function was studied in 11 patients treated with the implantable atrial defibrillator. Results of this study suggested that prompt cardioversion of spontaneous episodes of atrial fibrillation within 48 hours after onset was associated with early resolution of left atrial mechanical dysfunction seen after cardioversion. PMID- 11053722 TI - Left ventricular diastolic dysfunction in lone atrial fibrillation determined by Doppler tissue imaging of mitral annular motion. AB - In this study, we sought evidence for an underlying atrial or ventricular myopathy in patients with paroxysmal lone atrial fibrillation using standard echocardiographic parameters in addition to Doppler tissue imaging of mitral annular motion. No impairment in atrial contractile function was found, but there was evidence for impaired diastolic function in these patients. PMID- 11053723 TI - Comparison of effectiveness of an 8-mm versus a 4-mm tip electrode catheter for radiofrequency ablation of typical atrial flutter. AB - An 8-mm catheter does not appear superior to 4-mm tip electrode for atrial flutter ablation. The potential advantage of allowing higher energy delivery on a larger surface is compensated by the lack of consistent contact with the endocardial surface. PMID- 11053724 TI - Effect of concomitant digoxin and carvedilol therapy on mortality and morbidity in patients with chronic heart failure. AB - We retrospectively performed stepwise logistic regression analysis on 1,509 patients with chronic heart failure in 4 multicenter United States studies and 1 Australia-New Zealand study to examine the effect of digoxin in patients randomized to carvedilol or placebo. Patients receiving digoxin had more advanced heart failure, the incidence of hospitalization for any cause and the combination of all-cause death and all-cause hospitalization were the same in the digoxin versus no-digoxin groups. PMID- 11053725 TI - Plasma levels of A- and B-type natriuretic peptides in patients with hypertrophic cardiomyopathy or idiopathic dilated cardiomyopathy. AB - We investigated the relation between left ventricular structure and the secretion patterns of A- and B-type natriuretic peptides (ANP and BNP) by comparing their plasma levels in patients with hypertrophic cardiomyopathy (HC) and patients with idiopathic dilated cardiomyopathy (IDC). The secretion of ANP and BNP was much higher in patients with HC than in those with IDC; this shows that left ventricular cavity size is a key factor that regulates the secretion of ANP and BNP. PMID- 11053726 TI - Left ventricular mass and systolic function in human immunodeficiency virus infected patients after weight loss. AB - Transthoracic echocardiography was performed on 27 patients with human immunodificiency virus after weight loss and in 20 lean controls. Left ventricular mass index was significantly higher and left ventricular fractional shortening was significantly lower in patients with human immunodificiency virus after weight loss than in lean, normal controls. PMID- 11053727 TI - Predictors of left ventricular outflow obstruction following single-stage repair of interrupted aortic arch and ventricular septal defect. AB - This study looked at echocardiographic predictors of left ventricular outflow obstruction after primary neonatal repair of interrupted aortic arch and ventricular septal defect. Results of this study indicate that the only significant independent predictor of left ventricular outflow obstruction is aortic valve diameter; all patients with an aortic valve diameter <4.5 mm (Z score <-5) subsequently developed obstruction, whereas patients with annuli >4.5 mm (Z score >-5) remained free from obstruction. PMID- 11053728 TI - Comparison of mechanical properties of the left ventricle in patients with severe coronary artery disease by nonfluoroscopic mapping versus two-dimensional echocardiograms. AB - In 40 patients, we compared linear local shortening assessed with nonfluoroscopic electromechanical mapping as a function of regional wall motion with echocardiographic data in a subset of patients with severe coronary artery disease and subsequently decreased left ventricular function. Our study showed that nonfluoroscopic electromechanical mapping can accurately assess regional wall motion. In addition, this study showed a significant decrease in unipolar voltages among segments with declining regional function. PMID- 11053729 TI - Successful use of a biphasic waveform automated external defibrillator in a high risk child. PMID- 11053730 TI - Measuring growth of a phenanthrene-degrading bacterial inoculum in soil with a quantitative competitive polymerase chain reaction method. AB - We measured growth of a phenanthrene-degrading bacterium, Arthrobacter, strain RP17, in Forbes soil, amended with 500 ug g(-1) phenanthrene using a quantitative competitive polymerase chain reaction method. The inoculum, which was not indigenous to Forbes soil, grew from 5.55x10(5) colony forming units (cfu) g(-1) to 1.97x10(7) cfu g(-1) within 100 h after the cells were added to the soil. Maximum population density was reached before the highest degradation rate was observed 150 h after the cells were added to soil. Population density remained stable even after 56% of the phenanthrene had mineralized. This study is one of the few documented examples of growth by a non-indigenous bacterium in a non sterile soil amended with a pollutant. PMID- 11053731 TI - Characterization of halotolerant rhizobia isolated from root nodules of Canavalia rosea from seaside areas. AB - Twelve nodule isolates from Canavalia rosea, an indigenous leguminous halophyte growing in the seaside areas of southern Taiwan, were effective symbionts for the original host and able to grow at NaCl concentrations up to 3-3.5% (w/v). The taxonomy of these isolates was investigated using a polyphasic approach, including phenotypic characteristics, banding patterns of total proteins from sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), genomic fingerprint patterns from random amplified polymorphic DNA (RAPD) analysis, pulsed-field gel electrophoresis (PFGE) analysis, amplified 16S rDNA restriction analysis (ARDRA), 16S rRNA gene sequencing, and nifH gene sequencing. Based on the SDS-PAGE, RAPD, PFGE and ARDRA results, the 12 isolates are highly diverse. The 16S rRNA and nifH gene sequences were determined for isolates with distinct ARDRA patterns and compared with other members of the rhizobial species. We propose these isolates should be classified into the genus Sinorhizobium and distinguished from the current species of this genus. PMID- 11053732 TI - Symbiotic spirochetes in the termite hindgut: phylogenetic identification of ectosymbiotic spirochetes of oxymonad protists. AB - Some species of protists inhabiting the hindgut of lower-termites have a large number of ectosymbiotic spirochetes on the cell surface. The phylogenetic positions of the ectosymbiotic spirochetes of three oxymonad protists, Dinenympha porteri in the gut of Reticulitermes speratus, and Pyrsonympha sp. and Dinenympha sp. in Hodotermopsis sjoestedti, were investigated without cultivation of these organisms. Protist fractions carefully collected with a micromanipulator were used as templates for the amplification of small subunit ribosomal RNA genes (SSU rDNA). The phylogenetic tree inferred from the nucleotide sequences of the SSU rDNA showed that they were affiliated with the Treponema cluster of spirochetes and they were divided into two clusters. One was grouped together with the spirochetal sequences reported previously from the gut of termites and the other was related to the Treponema bryantii subgroup of treponemes (denoted as termite Treponema clusters I and II, respectively). Whole-cell in situ hybridization using a fluorescent-labeled oligonucleotide probe specific for the group of sequences in cluster II identified most of the ectosymbiotic spirochetes of the oxymonad protists in the gut of R. speratus and H. sjoestedti. However, not all of the ectosymbiotic spirochetes could be detected by means of this cluster II group-specific probe and the population of ectosymbiotic spirochetes of cluster II was different among the oxymonad species. In the case of D. porteri, an oligonucleotide probe specific for one member of cluster II recognized a portion of the ectosymbiotic spirochetes of cluster II, and their population was also different depending on the cell-type of D. porteri in terms of the attachment of ectosymbiotic spirochetes. The results indicate that the spirochetes of cluster II and probably those of a part of cluster I can be assigned to ectosymbiotic species of oxymonad protists and that the population of ectosymbiotic spirochetes associated with a single protist consists of at least three species of phylogenetically distinct spirochetes. PMID- 11053733 TI - Selective inhibition of reactions involved in methanogenesis and fatty acid production on rice roots. AB - Washed excised roots of rice (Oryza sativa) produced H(2), CH(4) and fatty acids (millimolar concentrations of acetate, propionate, butyrate; micromolar concentrations of isovalerate, valerate) when incubated under anoxic conditions. Surface sterilization of the root material resulted in the inactivation of the production of CH(4), a strong reduction of the production of fatty acids and a transient (75 h) but complete inhibition of the production of H(2). Radioactive bicarbonate was incorporated into CH(4), acetate, propionate and butyrate. About 20-40% of the fatty acid carbon originated from CO(2) reduction. In the presence of phosphate, CH(4) was exclusively produced from H(2)/CO(2), since phosphate selectively inhibited acetoclastic methanogenesis. Acetoclastic methanogenesis was also selectively inhibited by methyl fluoride, while chloroform or 2 bromoethane sulfonate inhibited CH(4) production completely. Production of CH(4), acetate, propionate and butyrate from H(2)/CO(2) was always exergonic with Gibbs free energies <-20 kJ mol(-1) product. Chloroform inhibited the production of acetate and the incorporation of radioactive CO(2) into acetate. Simultaneously, H(2) was no longer consumed and accumulated, indicating that acetate was produced from H(2)/CO(2). Chloroform also resulted in increased production of propionate and butyrate whose formation from CO(2) became more exergonic upon addition of chloroform. Nevertheless, the incorporation of radioactive CO(2) into propionate and butyrate was inhibited by chloroform. The accumulation of propionate and butyrate in the presence of chloroform probably occurred by fermentation of organic matter, rather than by reduction of acetate and CO(2). [U-(14)C]Glucose was indeed converted to acetate, propionate, butyrate, CO(2) and CH(4). Radioactive acetate, CO(2) and CH(4) were also products of the degradation of [U (14)C]cellulose and [U-(14)C]xylose. Addition of chloroform and methyl fluoride did not affect the product spectrum of [U-(14)C]glucose degradation. The application of combinations of selective inhibitors may be useful to elucidate anaerobic metabolic pathways in mixed microbial cultures and natural microbial communities. PMID- 11053734 TI - Fitness in soil and rhizosphere of Pseudomonas fluorescens C7R12 compared with a C7R12 mutant affected in pyoverdine synthesis and uptake. AB - Fluorescent pseudomonads have evolved an efficient strategy of iron uptake based on the synthesis of the siderophore pyoverdine and its relevant outer membrane receptor. The possible implication of pyoverdine synthesis and uptake on the ecological competence of a model strain (Pseudomonas fluorescens C7R12) in soil habitats was evaluated using a pyoverdine minus mutant (PL1) obtained by random insertion of the transposon Tn5. The Tn5 flanking DNA was amplified by inverse PCR and sequenced. The nucleotide sequence was found to show a high level of identity with pvsB, a pyoverdine synthetase. As expected, the mutant PL1 was significantly more susceptible to iron starvation than the wild-type strain despite its ability to produce another unknown siderophore. As with the wild-type strain, the mutant PL1 was able to incorporate the wild-type pyoverdine and five pyoverdines of foreign origin, but at a significantly lower rate despite the similarity of the outer membrane protein patterns of the two strains. The survival kinetics of the wild-type and of the pyoverdine minus mutant, in bulk and rhizosphere soil, were compared under gnotobiotic and non-gnotobiotic conditions. In gnotobiotic model systems, both strains, when inoculated separately, showed a similar survival in soil and rhizosphere, suggesting that iron was not a limiting factor. In contrast, when inoculated together, the bacterial competition was favorable to the pyoverdine producer C7R12. The efficient fitness of PL1 in the presence of the indigenous microflora, even when coinoculated with C7R12, is assumed to be related to its ability to uptake heterologous pyoverdines. Altogether, these results suggest that pyoverdine mediated iron uptake is involved in the ecological competence of the strain P. fluorescens C7R12. PMID- 11053735 TI - Identification of novel Archaea in bacterioplankton of a boreal forest lake by phylogenetic analysis and fluorescent in situ hybridization(1). AB - We report here on novel groups of Archaea in the bacterioplankton of a small boreal forest lake studied by the culture-independent analysis of the 16S rRNA genes amplified directly from lake water in combination with fluorescent in situ hybridization (FISH). Polymerase chain reaction products were cloned and 28 of the 160 Archaea clones with around 900-bp-long 16S rRNA gene inserts, were sequenced. Phylogenetic analysis, including 642 Archaea sequences, confirmed that none of the freshwater clones were closely affiliated with known cultured Archaea. Twelve Archaea sequences from lake Valkea Kotinen (VAL) belonged to Group I of uncultivated Crenarchaeota and affiliated with environmental sequences from freshwater sediments, rice roots and soil as well as with sequences from an anaerobic digestor. Eight of the Crenarchaeota VAL clones formed a tight cluster. Sixteen sequences belonged to Euryarchaeota. Four of these formed a cluster together with environmental sequences from freshwater sediments and peat bogs within the order Methanomicrobiales. Five were affiliated with sequences from marine sediments situated close to marine Group II and three formed a novel cluster VAL III distantly related to the order Thermoplasmales. The remaining four clones formed a distinct clade within a phylogenetic radiation characterized by members of the orders Methanosarcinales and Methanomicrobiales on the same branch as rice cluster I, detected recently on rice roots and in anoxic bulk soil of flooded rice microcosms. FISH with specifically designed rRNA-targeted oligonucleotide probes revealed the presence of Methanomicrobiales in the studied lake. These observations indicate a new ecological niche for many novel 'non extreme' environmental Archaea in the pelagic water of a boreal forest lake. PMID- 11053736 TI - Geostatistical analysis of the distribution of NH(4)(+) and NO(2)(-)-oxidizing bacteria and serotypes at the millimeter scale along a soil transect. AB - Soil is known to be heterogeneous for different activities at several spatial scales. Most studies have focused on macro- and meso-scales but micro-scales are rarely addressed. Hence, the spatial structure of NH(4)(+)- and NO(2)(-) oxidizers and of various serotypes of the latter was studied along two transects of approximately 10 cm, with two micro-samples taken from each millimeter. The presence of NH(4)(+)- and NO(2)(-)-oxidizers in a micro-sample was detected using colorimetric tests for the presence or absence of NO(2)(-) in cultures of the micro-samples. Geostatistics was used to determine the range of spatial influence of the bacterial types. For both types, semi-variograms indicated a non-random spatial pattern. The spatial dependence ranged from 2 to 4 mm for NO(2)(-)- and NH(4)(+)-oxidizers respectively, and the two bacterial types were not independently spatially located. Among the six serotypes of NO(2)(-)-oxidizers, only one exhibited a spatial dependence. The existence of a spatial structure at the millimeter scale suggests that micro-scale sampling should be employed for soil studies. Therby, data on bacterial populations and activities can be referred to a spatial scale which is meaningful to these organisms. PMID- 11053737 TI - Use of a chiA probe for detection of chitinase genes in bacteria from the Chesapeake Bay(1). AB - PCR primers specific for the chiA gene were designed by alignment and selection of highly conserved regions of chiA sequences from Serratia marcescens, Alteromonas sp., Bacillus circulans and Aeromonas caviae. These primers were used to amplify a 225 bp fragment of the chiA gene from Vibrio harveyi to produce a chiA gene probe. The chiA PCR primers and probe were used to detect the presence of the chiA gene in an assemblage of 53 reference strains and gave consistent results. Selected chiA fragments amplified by PCR were cloned and sequenced from nine known strains and from Chesapeake Bay isolates 6d and 11d. This confirmed the specificity and utility of the primers for detection of chiA-positive environmental strains. Over 1000 bacterial isolates from Chesapeake Bay water samples were tested for the presence of the chiA gene which was found to be present in 5-41% (average 21%) of the culturable bacterial community. The approach developed in this study was valuable for isolation and enumeration of chiA-positive bacteria in environmental samples. PMID- 11053738 TI - Detection of Desulfotomaculum in an Italian rice paddy soil by 16S ribosomal nucleic acid analyses. AB - Two specific primers were developed for the amplification of 16S rRNA genes of Desulfotomaculum lineage 1 to detect members of the genus Desulfotomaculum in rice field soil. The combination of both primers in PCR allowed the specific amplification and cloning of ten 16S rDNA sequences of this group from rice paddy soil DNA extracts. The phylogenetic analysis showed that these sequences formed a deeply branching cluster within Desulfotomaculum lineage 1, together with two sequences from the database and two sequences from a hydrocarbon-contaminated aquifer. Dissimilarity values to validly described species, including recently isolated strains of Desulfotomaculum from rice paddy microcosms, were higher than 12%. Within the new cluster the cloned sequences formed three separate groups which were each represented by at least two sequences with identities of >/=99% while one sequence represented an additional group. The sequences should represent sulfate-reducing organisms because they clearly fell into the physiologically coherent group of Gram-positive sulfate reducers. The relative abundance of bacteria of the Desulfotomaculum lineage 1 in rice paddy soil and root samples was estimated with rRNA dot blot hybridizations of extracted RNA. The relative RNA content of Desulfotomaculum lineage 1 was 0.55% in the bulk soil and 1% in the rice root samples, respectively, of the total 16S rRNA content (probe Eub338). Hybridization of rRNA with a probe targeting the new cluster represented by the cloned sequences confirmed the high abundance of 16S rRNA sequences from this cluster in the rice paddy field samples. Another hybridization probe detecting Desulfotomaculum acetoxidans and two closely related Desulfotomaculum isolates from rice paddy soil indicated that these bacteria were less abundant. PMID- 11053739 TI - Detection and distribution of insertion sequence 1 (IS1)-containing bacteria in the freshwater environment(1). AB - The distribution of insertion sequence 1 (IS1)-containing bacteria was investigated in Windermere (Cumbria, UK), a freshwater body impacted by treated sewage discharge and run-off from the surrounding catchment. Culturable IS1 containing bacteria were recovered from the water column at three depths in Windermere North Basin (WNB) and South Basin (WSB), and from sediment at both sites (at the sediment surface in WSB and to a depth of 12-13 cm in WNB). Polymerase chain reaction amplification of IS1 and the Escherichia coli/Shigella sp. specific gene uidA, from community DNA from shallow sediments, extended the detection limit beyond that of culture at both sites. This detection was extended further into deep sediment extracted from WNB as IS1 and uidA were detected in sub-samples to a depth of 4.7 and 2.3 m, respectively. Analysis of a representative subset of 90 IS1-carrying isolates recovered from water and sediment at both sites demonstrated 21 heterogeneous IS1 profiles with estimated copy numbers ranging from 1 to 16. Identification of the host bacteria showed that the element was confined mainly to Enterobacter spp. However, this study showed IS1 to be present in Citrobacter freundii for the first time. Plasmids were carried by 75.3% of enterobacterial isolates and four plasmids (2.6%) carried IS1. DNA sequence analysis of five IS1 clones demonstrated that IS1 isoforms from this study were similar (>89% nucleotide identity) to known IS1 isoforms. Two isoforms of IS1 from a single Enterobacter cloacae isolate differed by 6.7% at the nucleotide level suggesting that they had been acquired independently. PMID- 11053740 TI - Three-dimensional echocardiography paves the way toward virtual reality. AB - The heart is a three-dimensional (3-D) object and, with the help of 3-D echocardiography (3-DE), it can be shown in a realistic fashion. This capability decreases variability in the interpretation of complex pathology among investigators. Therefore, it is likely that the method will become the standard echocardiography examination in the future. The availability of volumetric data sets allows retrieval of an infinite number of cardiac cross-sections. This results in more accurate and reproducible measurements of valve areas, cardiac mass and cavity volumes by obviating geometric assumptions. Typical 3-DE parameters, such as ejection fraction, flow jets, myocardial perfusion and LV wall curvature, may become important diagnostic parameters based on 3-DE. However, the freedom of an infinite number of cross-sections of the heart can result in an often-encountered problem of being "lost in space" when an observer works on a 3-DE image data set. Virtual reality computing techniques in the form of a virtual heart model can be useful by providing spatial "cardiac" information. With the recent introduction of relatively low cost portable echo devices, it is envisaged that use of diagnostic ultrasound (US) will be further boosted. This, in turn, will require further teaching facilities. Coupling of a cardiac model with true 3-D echo data in a virtual reality setting may be the answer. PMID- 11053741 TI - Reproducibility of calcified lesion quantification: a longitudinal intravascular ultrasound study. AB - In view of a prospective intravascular ultrasound (IVUS) study, the reproducibility of the extent of the calcified lesion in IVUS images derived from separate pull-back maneuvers was assessed. Patients (n = 34) were imaged with IVUS before and after percutaneous transluminal angioplasty (PTA) and at 1-y follow-up. In the presence of a calcified lesion, the largest arc and the length of the matched calcified lesions was assessed. Interobserver differences in arc measurements were low (< or = 0.7%), with low coefficients of variation (< or = 5.8%). Similarly, interexamination differences in arc and length measurements were small (< or = 1.1%), with low coefficients of variation (< or = 3.2%). At follow-up, a nonsignificant increase in both the arc (1.9%) and length (1.7%) of the calcified lesion was observed. This study showed that measurements of the calcified lesion are highly reproducible; changes seen at 1-y follow-up were not significant. We conclude that IVUS may be used to monitor the effect of medical intervention on the extent of the calcified lesion in a longitudinal study. PMID- 11053742 TI - Ultrasonographic assessment for response to radiochemotherapy of metastatic cervical lymph nodes in head and neck cancer: usefulness of grey-scale and color doppler sonography. AB - To predict the response of lymph node metastasis to preoperative radiochemotherapy sonographically, the correlation between ultrasonographs and histologic features was retrospectively examined in 43 metastatic cervical lymph nodes from 24 patients with squamous cell carcinoma in the oral and maxillofacial region. Ultrasonographs were compared among poor-, good-, and complete-response lymph nodes. Before radiochemotherapy, hypoechoic internal echo and intranodal blood perfusion demonstrated many complete-response nodes; in contrast, most poor response nodes showed peripheral blood perfusion and an avascular pattern, but did not have specific internal echo intensity. Complete-response nodes showed a significant reduction in their maximum and minimum diameters after radiochemotherapy. These results indicate that ultrasonography is useful for predicting the response of cervical lymph node metastasis to radiochemotherapy. PMID- 11053743 TI - Pathological angioarchitecture in lymph nodes: underlying histopathologic findings. AB - Pathologic changes of the intranodal angioarchitecture, as displayed by colour Doppler sonography, were used in recent studies to predict malignant infiltration of superficial lymph nodes. We searched for the underlying histopathologic findings in a prospective study including 100 lymph nodes in 86 patients. In the histopathologic specimens, we evaluated tumour infiltration, distribution of intranodal vessels, hilar structures, thickness of the capsule and tissue alterations (necrosis, sclerosis, lipo-/fibromatosis). Using the only prospectively tested classification, a pathologic angioarchitecture was described as displacement, aberrant course, avascular foci or subcapsular vessels. Testing these four criteria by multivariate regression analysis, the most important correlations were found between the following pairs of sonographic and histopathologic findings: displacement with perinodal tumour spread, aberrant vessels with intranodal sclerosis, avascular foci with intranodal necroses and subcapsular vessels with intranodal necroses. In conclusion, the criteria describing a pathologic angioarchitecture correlate with histopathologic findings that are frequently seen in malignant lymph nodes. PMID- 11053744 TI - Tissue Doppler imaging in detection of myocardial dysfunction in survivors of childhood cancer treated with anthracyclines. AB - The applicability of tissue Doppler imaging (TDI) was investigated for estimating cardiac function in long-term survivors of childhood cancer treated with anthracyclines. A total of 63 children (age range 7.8-17.3 y) underwent standard echo Doppler cardiographic studies of blood flow velocities, left ventricular dimensions and fractional shortening, followed by measurements of peak myocardial velocities and direction using the noninvasive tissue Doppler imaging (TDI) technique. All 63 were late survivors (median 7.1 y, range 3.5-13.5 y after end of therapy) who had received mean (+/- SD) cumulative dose of 242 (+/- 141) mg/m(2) of anthracyclines. The control group consisted of 160 healthy subjects (age range 4 to 17.9 y). Standard echo-Doppler anatomical parameters that were found significantly (p < 0.01) different for the study group are: RV wall thickness (decreased); LV diameter (increased); and LV fractional shortening (decreased). Studied hemodynamic parameters were not found to be different between the two groups. Quantitative TDI parameters: peak late diastolic myocardial velocities, as well as transmyocardial systolic and diastolic velocity differences, were significantly lower in late survivors than in the healthy pediatric population (p < 0.01). Qualitative local functional impairment of the movement of the left ventricular walls was detected in 20% of the patients. TDI might become a useful noninvasive method for detecting subclinical myocardial damage in apparently healthy children who received moderate doses of anthracyclines for treatment of childhood malignancy. Prospective studies with TDI for the detection of regional myocardial abnormalities are recommended. PMID- 11053745 TI - Contrast-enhanced power Doppler harmonic imaging--influence on visualization of renal vasculature. AB - To compare contrast-enhanced power Doppler (PD) harmonic imaging (CHI) with contrast-enhanced power Doppler fundamental imaging (CPD) in the depiction of renal cortical vessels, 20 healthy volunteers were subjected to PD imaging and HI assessment of the kidney after bolus injection of Levovist(R) (SH U 508A). System settings were standardized and the pulse-repetition frequencies (PRF) systematically toggled from 750 to 500 and 250. Videotapes were independently reviewed by three readers with regard to the presence of artefacts, the degree of Doppler signal enhancement, demarcation of vessels and the extent of visualization. The assessments were graded separately for each PRF in accordance with a multistage scoring system. In comparison to contrast-enhanced PD, artefacts were significantly lower with CHI for all PRF (p = 0.0001). Vessels were better visualized (p = 0.002) and less blurred (p = 0.006) with CHI than with CPD. There was no significant difference in the extent of Doppler signal increase between CPD and the contrast-enhanced harmonic mode. Combination of the contrast-enhanced harmonic method and PD allows the PRF to be lowered and, by balancing the greater susceptibility of PD to interference from clutter, increases the likelihood of detection of flow in small vessels. PMID- 11053746 TI - The DopFet system: a new ultrasonic Doppler system for monitoring and characterization of fetal movement. AB - To enable the investigation of fetal movement in a manner similar to fetal heart rate (FHR) monitoring we have developed an apparatus (the DopFet system) that consists of a pair of miniature sensors, a 2-MHz continuous-wave directional Doppler electronic module and a laptop personal computer. One of the sensors is aimed at the fetal limbs and the other at the thorax to detect heart and upper body movements. The signals are analyzed, presented in real-time and postprocessed by software developed by us. The postprocessing software computes a number of parameters (the DopFet parameters) describing fetal movement. These parameters can be divided into two categories: parameters that describe the quantity of fetal movement (i.e., number of movements) and parameters that describe qualitative aspects of fetal movement (i.e., average movement duration). Future studies using the DopFet system will be aimed at discovering which of these parameters or combination of parameters is the best indicator of fetal well being. We present an example of a 0.5 h recording and the results of testing on 23 volunteer mothers. These results show good sensitivity of the system compared to real-time ultrasound (US). The system detects 96% of rolling movements, 100% of flexion movements and 97% of leg movements. PMID- 11053747 TI - Errors in ultrasound digital image distance measurements. AB - Digital image distance measurements have two sources of error: they are the inherent image pixelation error and modality-specific registration errors. A theoretical analysis was performed to determine the pixelation error uncertainty in ultrasound (US) clinical and phantom digital image distance measurements. For horizontal or vertical phantom image distance measurements, the average absolute image pixelation error was found to be 0.5 pixel width. In the clinical horizontal case, the combined absolute distance measurement error due to pixel size and operator cursor placement errors was found to be 1.5 pixel widths (for a cursor placement error of 1 pixel width). For image clinical distance measurements, a formula is given for the dependence of the measurement error on the inclination of the measured structure in the pixel matrix. A protocol is recommended that minimizes all sources of error in clinical image distance measurements. PMID- 11053748 TI - The effect of acoustic velocity on phantom measurements. AB - Urethane rubber ultrasound (US) phantoms have a much lower acoustic velocity (1430-1450 m/s) than the accepted soft tissue average of 1540 m/s. Two important questions arise: can the rod positions in these rubber phantoms be adjusted so that they may be used to test equipment distance measurement accuracy for all types of multielement transducers, and can they be used to measure beam focus (using the spread of the rod blur patterns)? These questions were addressed for linear-, phased-, convex- and vector-array transducers. Theoretical predictions for the different transducers' distance measurement errors agreed with careful measurements obtained with a specially designed array of stainless-steel rods immersed in paraffin oil (1447 m/s). The conclusions of this study are that phantoms with acoustic velocities different from 1540 m/s cannot be used to check distance measurement accuracies of all the types of real-time transducers, nor to predict a transducer's focusing performance in clinical scans. PMID- 11053749 TI - Transcranial Doppler blood velocity measurement--the effect of changes in velocity profile. AB - Transcranial Doppler (TCD) units measure blood velocity in the middle cerebral artery (MCA) and are used to examine the effects of pharmacological agents. The units actually measure the average of the maximum blood velocity envelope (aveV(max)) and it is assumed that changes in aveV(max) follow changes in the true mean velocity (aveV(mean)). This may not be true if there are changes in velocity profile. Results from previous TCD studies using acetazolamide (ACZ) and caffeine were examined for evidence for changes in velocity profile. ACZ increased aveV(max) by 21% (95% CI 13 to 29%) and aveV(mean) by 14% (95% CI 9 to 19%). Caffeine decreased aveV(max) by 8% (95% CI 4 to 12%) and aveV(mean) by 5% (95% CI 4% increase to 13% decrease). In both cases, the true change, measured using aveV(mean) was lower, indicating possible changes in velocity profile. We conclude that the possibility of changes in velocity profile must be considered when using TCD to quantify changes in blood velocity. PMID- 11053750 TI - Cavitational mechanisms in ultrasound-accelerated thrombolysis at 1 MHz. AB - Inertial cavitation is hypothesized to be a mechanism by which ultrasound (US) accelerates the dissolution of human blood clots when the clot is exposed to a thrombolytic agent such as tissue plasminogen activator (t-PA). To test this hypothesis, radiolabeled fibrin clots were exposed or sham-exposed in vitro to 1 MHz c.w. US in a rotating sample holder immersed in a water-filled tank at 37 degrees C. Percent clot dissolution after 60 min of US exposure was assessed by removing the samples, centrifuging, and measuring the radioactivity of the supernatant fluid relative to the pelletized material. To suppress acoustic cavitation, the exposure tank was contained within a hyperbaric chamber capable of pneumatic pressurization to 10 atmospheres (gauge). Various combinations of static pressure (0, 2, 5, and 7.5 atm gauge), US (0 or 4 W/cm(2) SATA), and t-PA (0 or 10 microg/mL) were employed, showing statistically significant reductions in thrombolytic activity as static pressure increased. To gain further insight, an active cavitation detection scheme was employed in which 1-micros duration tonebursts of 20-MHz US (< 1 kPa peak negative pressure, 1 Hz PRF) were used to interrogate clots subjected to US and static pressure. Results of this cavitation detection scheme showed that scattering from within the clot and broadband acoustic emissions that were both present during insonification were significantly reduced with application of static pressure. However, only about half of the acceleration of thrombolysis due to US could be removed by static pressure, suggesting the possibility of other mechanisms in addition to inertial cavitation. PMID- 11053751 TI - Stimulated acoustic emission: pseudo-Doppler shifts seen during the destruction of nonmoving microbubbles. AB - The purpose of this study was to evaluate the appearance and the characteristics of stimulated acoustic emission (SAE) as an echo contrast-specific color Doppler phenomenon with impact on myocardial contrast echocardiography (MCE). Stationary microbubbles of the new contrast agent SH-U 563A (Schering AG) were embedded within a tissue-mimicking gel material. Harmonic power Doppler imaging (H-PDI), color Doppler and pulse-wave Doppler data were acquired using an HDI-5000 equipped with a phased-array transducer (1.67/3.3 MHz). In color Doppler mode, bubble destruction resulted in random noise like Doppler signals. PW-Doppler revealed short "pseudo-Doppler" shifts with a broadband frequency spectrum. Quantification of SAE events by H-PDI demonstrated an exponential decay of signal intensities over successive frames. A strong linear relationship was found between bubble concentration and the square root of the linearized H-PDI signal for a range of concentrations of more than two orders of magnitude (R = 0.993, p < 0.0001). Intensity of the H-PDI signals correlated well with emission power (R = 0.96, p = 0.0014). SAE results from disintegration of microbubbles and can be demonstrated by all Doppler imaging modalities, including H-PDI. Intensity of SAE signals is influenced by the applied acoustic power and correlates highly with the concentration of microbubbles. Because intensity of SAE signals correlates highly with echo contrast concentrations, analysis of SAE signals might be used for quantitative MCE. PMID- 11053752 TI - Experimental study of the effects of Optison concentration on sonoporation in vitro. AB - Lethal sonoporation and reparable sonoporation were observed in Jurkat lymphocytes in suspension with the addition of varying amounts of Optison, a commercially available bubble-based contrast agent. For given ultrasound (US) exposure conditions (spatial peak-pressure amplitude of 0.2 MPa, duty cycle 10% and 2-MHz frequency), sonoporation was directly related to the bubble-to-cell ratio (in a range from 0 to 230). It was found that the nearest bubble-cell spacing was also related to the occurrence frequency of bioeffects. A constant bubble-to-cell ratio often provided very different results for two different initial cell concentrations (200,000 cells/mL and 600,000 cells/mL), with the higher cell concentration generally exhibiting higher levels of sonoporation. In contrast, a constant bubble-to-cell spacing provided similar results between the two initial cell concentrations. The frequency of reparable and lethal sonoporation was seen to decay as the inverse-cube power of the nearest bubble cell spacing. Significant reparable sonoporation was observed at a bubble-cell spacing that was 10 microm larger than the minimum spacing at which significant lethal sonoporation was observed. Preliminary analysis also suggests the possibility of a step-wise increase in lethal sonoporation as spacing decreases; further experiment is needed. PMID- 11053753 TI - Acoustic droplet vaporization for therapeutic and diagnostic applications. AB - A phase shift droplet emulsion is introduced as an aid to unusual ultrasound (US) applications. The transpulmonary droplet emulsion (90% < 6 microm diameter) is made by mixing saline, bovine albumin and dodecafluoropentane. It has been observed that an acoustic pressure threshold exists, above which the droplets vaporize into bubbles approximately 25 times the original diameter. For frequencies between 1.5 and 8 MHz, the threshold decreases from 4.5 to 0.75 MPa peak rarefactional pressure. This paper presents preliminary results for droplet preparation and their evaporation as a function of applied acoustic pressure and frequency, as well as simulations of the lifetime of these gas bubbles based on gas diffusion. In vivo experiments were simulated by the evaporation of droplets in blood flowing under attenuating material. We propose that this agent might be useful for tissue occlusion in cancer treatment, as well as for phase aberration corrections in acoustic imaging. PMID- 11053754 TI - Physical properties of the mitral valve tissue assessed by tissue sound speed in cardiac amyloidosis: relationship to the severity of mitral regurgitation. AB - Cardiac amyloidosis has been documented to show mitral regurgitation (MR) and a thickened mitral valve (MV) due to amyloid deposits. However, the changes in the physical properties of the thickened MV tissue in cardiac amyloidosis, which may be a causative factor of the MR, have not been described. Physical properties of the tissue, which are expressed by the elastic bulk modulus, can be evaluated by tissue sound speed. If biological tissue is assumed to be fluid-like, the tissue sound speed may be given by c= square root of K/rho, where c is the tissue sound speed, K is the elastic bulk modulus, and rho is the density. A reduction in tissue sound speed indicates a reduction in the elastic bulk modulus of the tissue, assuming that there is little change in rho. This suggests that the tissue is less elastic. The purpose of this study was to assess the physical properties of MV tissue by evaluating the sound speed of the MV tissue in cardiac amyloidosis. MV specimens were obtained at autopsy from 20 control adults without cardiovascular diseases and from 20 patients with cardiac amyloidosis. An acoustic microscope operating at 450 MHz was used to measure the tissue sound speed in the tip and basal portions of the MV tissue. The density of MV tissue was measured by microgravimetry. The severity of the MR had been evaluated by Doppler echocardiography before death, and it was compared with the tissue sound speed measured after death. In cardiac amyloidosis showing mild MR, the tissue sound speed of the MV in the tip portion (1605 +/- 19 m/s) and in the basal portion (1791 +/- 64 m/s) were lower than the corresponding values in control subjects (1637 +/- 42 m/s and 1851 +/- 62 m/s). However, these differences were not statistically significant. In cardiac amyloidosis showing moderate MR, the tissue sound speed of MV in the tip portion (1563 +/- 17 m/s) and in the basal portion (1654 +/- 59 m/s) were significantly lower than the corresponding values in the control subjects (p < 0.001) and the patients with mild MR (p < 0.05). No significant differences were observed in the density of MV tissue among the three groups. Therefore, the low value of the MV tissue sound speed in patients with cardiac amyloidosis indicated a reduced elastic bulk modulus, suggesting the less elasticity of the MV tissue. Furthermore, the patients with moderate MR demonstrated the greater reduction in the tissue sound speed than the patients with mild MR. The data suggest that the changes in physical properties of the MV tissue may be one of the causes of MR in cardiac amyloidosis. PMID- 11053755 TI - Concerning physiological interpretation of an impedance index derived from transcranial Doppler study. PMID- 11053756 TI - Response from dr. giller PMID- 11053758 TI - The more drinking, the more fun; but is there a calculus of fun, and should it drive policy? PMID- 11053757 TI - Exploring psychological benefits associated with moderate alcohol use: a necessary corrective to assessments of drinking outcomes? AB - The aim of this paper is to identify positive psychological concomitants of moderate alcohol consumption. Current research and public-health perspectives on alcohol emphasize harms disproportionately relative to benefits. The major exception is research establishing beneficial effects of moderate drinking on cardiovascular health and overall mortality. In addition, much observational and experiential data suggest the widespread prevalence of positive drinking experiences. This paper is one of the first attempts since 1985 to codify such benefits in epidemiological terms. Methodological difficulties in accomplishing this include defining moderate drinking, controlling for confounding variables, and establishing causality. Nonetheless, evidence of psychological benefits has been found in experimental, observational, interview, self-report, correlational, and some prospective research. These positive findings are in the areas of subjective health, mood enhancement, stress reduction, sociability, social integration, mental health, long-term cognitive functioning, and work income/disability. Problem drinkers and alcoholics also seek mood and other benefits from alcohol, but are more likely to drink to counteract negative feelings and to support their egos than are social drinkers. It is as yet impossible to determine to what extent moderate alcohol consumption causes positive psychological outcomes and to what extent it is part of a complex pattern of mutually reinforcing variables. PMID- 11053759 TI - Clusters of drug involvement in Panama: results from Panama's 1996 National Youth Survey. AB - This report is based on the first epidemiological investigation of clustering of tobacco, alcohol, inhalant, and other drug involvement within individual schools using data from Panama's 1996 National Youth Survey on Alcohol and Drug Use. Clustering was estimated with the Alternating Logistic Regression method. Adjusted estimates of pair-wise cross-product ratios (PWCPR), a measure of clustering, show modest clustering (i.e. PWCPR>1.0) at the school level for tobacco smoking (PWCPR=1.41; 95% confidence interval, CI=1.22-1.64), alcohol consumption (PWCPR=1.33; 95% CI=1.22-1.45), use of inhalants, (PWCPR=1.35; 95% CI=1.07-1.69), and other drug use (PWCPR=1.38; 95% CI=1.14-1.68). These findings provide preliminary evidence that the odds of drug use among school-attending youths increase when another youth in the same school uses drugs, and suggest a new line of research on within-school diffusion that should include the identification of school-level factors that contribute to student drug use. PMID- 11053760 TI - Needle sharing in opioid-dependent outpatients: psychological processes underlying risk. AB - Needle sharing contributes to the spread of the human immunodeficiency virus and other health concerns and remains a persistent problem among injection drug users. We determined whether needle sharing may be related to the discounting of the value of delayed outcomes. Outpatients in treatment for heroin dependence indicated preference for immediate versus delayed hypothetical monetary and heroin outcomes in a titration procedure that determined indifference points at various delays. The degree to which the delayed outcomes lost value was estimated with a nonlinear decay model. Participants who agreed to share a needle in a scenario (N=15) discounted delayed money more steeply than did the nonsharing group (N=17). Both groups discounted delayed heroin more steeply than delayed money. Persistent needle sharing may be related to the relative inability of delayed outcomes to impact current behavior. Training to mitigate the effect of delay on outcome value may offer reductions in needle sharing and drug abuse. PMID- 11053761 TI - The relationship between psychopathology and smoking cessation treatment response. AB - The study evaluated the relationship of psychopathology to treatment response of 208 smokers prescribed transdermal nicotine (8 weeks). Participants were relatively high functioning (DSM-IV axis V score) outpatients in a university based clinic. The primary study objective was to determine whether patients with a history of either a DSM-IV axis I or II diagnosis would have poorer during treatment response (patch adherence, smoking) and lower rates of smoking cessation at post-patch follow-up (study weeks 9, 26, 52) than those without a diagnosis. While there was some indication that patients with a history of psychopathology wore the patch less frequently, psychopathology was not associated with during- and post-treatment smoking. PMID- 11053762 TI - Patterns of improvement after methadone treatment: 1 year follow-up results from the National Treatment Outcome Research Study. AB - One year outcomes for substance use behaviours, health and criminal behaviour, and variation in treatment response, are reported for patients recruited to methadone maintenance and methadone reduction treatment programmes as part of NTORS. Significant reductions in the use of all illicit target drugs were found at follow-up for patients recruited to the methadone maintenance and methadone reduction modalities. Because of similarities in the treatments received by clients in the two modalities we caution against interpreting these findings as showing that methadone maintenance and reduction treatments lead to similar outcomes. At this stage, it is suggested that these outcomes be regarded as reflective of exposure to some general methadone substitution treatment. Further investigation of the outcomes for the two modalities will be conducted. Cluster analyses were used to classify patients according to level of improvement in drug use. Four groups were identified. Two groups (59% of cases) showed substantial reductions in their illicit drug use and criminality as well as reduced physical and psychological symptoms. Twenty two percent of cases showed poor outcomes across a range of measures. Results for alcohol consumption were less satisfactory for patients in all groups. A majority of patients achieved widespread improvements across a range of outcome measures after treatment in existing methadone treatment services. These changes represent important clinical benefits to the individual clients, to their families and to society. PMID- 11053763 TI - Theoretical and observed subtypes of DSM-IV alcohol abuse and dependence in a general population sample. AB - The purpose of this study was to quantify the degree of heterogeneity of the Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition (DSM-IV) alcohol abuse and dependence categories by comparing the number of theoretically predicted subtypes of each category with those observed in a nationally representative sample of the US general population. Among respondents classified with a past year diagnosis of abuse, only 11 (47.8%) of the 23 theoretically predicted subtypes of abuse were observed, while 53 (53.5%) of the 99 theoretically predicted subtypes of dependence were observed in this general population sample. Approximately 90% of the respondents classified with abuse could be represented by three subtypes of abuse and 70% of the respondents with current diagnoses of dependence could be characterized by six subtypes of dependence, indicating the relative homogeneity of both diagnostic categories. Sociodemographic differentials were also observed including the reduction in the number of observed subtypes of abuse and dependence with age as well as the larger numbers of subtypes associated with males and whites relative to females and blacks, respectively. Implications of these results are discussed in terms of increased physical morbidity and disruption of family life as persons with alcohol use disorders age, the potential role of physiological and impaired control over drinking indicators of dependence as critical features of the disorder, and the future need to examine the conceptual basis of the abuse category and to conduct longitudinal epidemiological research. PMID- 11053764 TI - Using cluster analysis of alcohol use disorders to investigate 'diagnostic orphans': subjects with alcohol dependence symptoms but no diagnosis. AB - The purpose of this paper is to examine the characteristics of community subjects with one or two alcohol dependence symptoms who did not satisfy the criteria for Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition (DSM-IV) alcohol dependence or abuse (sub-diagnostic group or diagnostic orphans). Variables not included in the DSM-IV alcohol abuse and dependence criteria are used to compare the different alcohol statuses. The results indicate that the sub diagnostic group 'diagnostic orphans' formed a cluster distinct from that of the non-problem drinkers group, and appeared to be closer to those with alcohol abuse than to those with alcohol dependence. The diagnosis of DSM-IV alcohol dependence (with three, four, or five or more symptoms) appeared to be its own entity. The findings lend credence to the requirement of three symptoms (in any 12 months) for the diagnosis of alcohol dependence in DSM-IV. However, those with one or two symptoms of alcohol dependence should be considered along with alcohol abuse as an entity in future DSM classifications. PMID- 11053765 TI - Analysis of the distribution of peripheral blood lymphocyte subsets associated with chronic ethanol treatment in rats with a disturbed circadian cycle. AB - The present study examined the composition of lymphoid subsets in the peripheral blood of alcohol-preferring (PRF) and non-preferring (NPF) rats, in an experimental model of alcoholism involving the disruption of the circadian cycle. The absolute and relative number of lymphocytes in specific subsets (CD3, CD4, CD8, NK, CD45RA) were measured using the flow cytometry method. When control animals with a disrupted circadian cycle (KN) were compared with a normal diurnal cycle group (KD), it was noticed that this disruption led to an increase in the absolute number of lymphocytes T (CD3(+), CD4(+) and CD8(+) cells) and lymphocytes B (CD45RA(+)). After the period of time when the alcohol preference was seen, there was a change in response - as measured by the numbers and the percentage of lymphoid subsets in NPF rats - involving a lowering of NK and CD45RA(+) cells. It seems that these animals exhibit higher sensitivity towards prolonged ethanol intoxication. However, the PRF animals - for whom the analysed values were close to those of the control group (KN) - tolerated the toxic effects of ethanol better and this may be related to their genetic predisposition. PMID- 11053766 TI - Gonadal hormone levels in injection drug users. AB - Levels of serum sex hormones, particularly testosterone, luteinizing hormone, and follicle-stimulating hormone, have been shown to change in various disease states. Hypogonadism has been well-documented in patients with HIV infection. As HIV infection and injection drug use are closely linked, we examined whether injection drug users (IDU's) exhibit hormone abnormalities. We evaluated men participating in the ALIVE study (AIDS Linked to Intravenous Experiences), a large cohort study conducted in Baltimore, MD. We found that 20% of 40 IDU subjects (20 HIV+, 20 HIV-) with a mean age 41.5+/-0.9, had low serum total testosterone concentration. We were unable to detect a direct correlation between drug use and hormone levels. Further study is needed on the hormonal milieu of the IDU patient. PMID- 11053767 TI - Drinking and heavy drinking by students in 18 countries. AB - This paper examines how levels of drinking, daily drinking and heavy drinking are related in 18 countries. There is wide national variation in consumption patterns. The proportions of students drinking and the proportion drinking at least six times in the past month correlated with per capita consumption in the total adult population. Some student drinking variables correlated with each other. Most countries (n=14) could be classified into one of two distinct groups based on the percentage of students who reported drinking at least six times per month, the percentage reporting drinking at least five drinks per day at any time in the last month and per capita alcohol consumption. Some countries in these groups were geographically close but others were not. Further research is needed to understand differences and similarities among countries with respect to student drinking. PMID- 11053768 TI - Drinking and smoking as concurrent predictors of illicit drug use and positive drug attitudes in adolescents. AB - The study investigates the relationship between smoking and drinking, and the use of illicit drugs in a cohort of London adolescents. A high prevalence of drug experimentation and positive attitudes to illicit drug use were characteristic of those who both drank alcohol and smoked cigarettes on a regular basis. There was then a clear hierarchy in which lower prevalence of use and more negative attitudes marked those who only smoked, then those who only drank, while non smokers and non-drinkers (the largest group) had lowest lifetime and recent drug use prevalence and the most negative attitudes about drug use. PMID- 11053769 TI - Erratum to 'Inter-rater reliability of the SCID alcohol and substance use disorders section among adolescents'. PMID- 11053770 TI - Chronic myocardial hypoxia increases nitric oxide synthase and decreases caveolin 3. AB - Nitric oxide synthase (NOS) is believed to play an important role in protecting the myocardium against ischemia. Chronic hypoxia from birth increases NOS activity in the myocardium resulting in enhanced nitric oxide production and increased resistance to ischemia. We examined the effects of chronic hypoxia on NOS gene and protein expression and on NOS protein association with caveolin-3. Rabbits were raised from birth in a normoxic (F(I)O(2) = 0.21) or a hypoxic (F(I)O(2) = 0.12) environment for 9 d, and then the hearts were isolated. Ribonuclease protection assays revealed that chronic hypoxia did not alter NOS transcript levels for NOS1, NOS2, or NOS3. The most abundant transcript was NOS3. Western analysis revealed NOS3 was the only isoform detected. Immunoblots of NOS3 immunoprecipitates showed that chronic hypoxia increases NOS3 protein by 2.0 +/- 0.4-fold and decreases the amount of caveolin-3 that can be coprecipitated with NOS3 by 5.5 +/- 0.9-fold. Immunoblots of normoxic and hypoxic hearts showed that chronic hypoxia decreases the amount of caveolin-3 in heart homogenates by 2. 2 +/- 0.5-fold. These data suggest that a decrease in caveolin-3 plays a role in the mechanisms by which chronic hypoxia increases NOS3 activity in the myocardium. PMID- 11053771 TI - BCL-2 protects peroxynitrite-treated thymocytes from poly(ADP-ribose) synthase (PARS)-independent apoptotic but not from PARS-mediated necrotic cell death. AB - In thymocytes, peroxynitrite induces poly(ADP-ribose) synthetase (PARS) activation, which results in necrotic cell death. In the absence of PARS, however, peroxynitrite-treated thymocytes die by apoptosis. Because Bcl-2 has been reported to inhibit not only apoptotic but also some forms of necrotic cell death, here we have investigated how Bcl-2 regulates the peroxynitrite-induced apoptotic and necrotic cell death. We have found that Bcl-2 did not provide protection against peroxynitrite-induced necrotic death, as characterized by propidium iodide uptake, mitochondrial membrane potential decrease, secondary superoxide production, and cardiolipin loss. In the presence of a PARS inhibitor, peroxynitrite-treated thymocytes from Bcl-2 transgenic mice showed no caspase activation or DNA fragmentation and displayed smaller mitochondrial membrane potential decrease. These data show that Bcl-2 protects thymocytes from peroxynitrite-induced apoptosis at a step proximal to mitochondrial alterations but fails to prevent PARS-mediated necrotic cell death. Activation of tissue transglutaminase (tTG) occurs in various forms of apoptosis. Peroxynitrite did not induce transglutaminase activity in thymocytes and did not have a direct inhibitory effect on the purified tTG. Basal tTG was not different in Bcl-2 transgenic and wild type cells. PMID- 11053772 TI - Acrolein, a product of lipid peroxidation, inhibits glucose and glutamate uptake in primary neuronal cultures. AB - Oxidative stress has been implicated in the pathogenesis of several neurodegenerative disorders including Alzheimer's disease (AD). Increased lipid peroxidation, decreased levels of polyunsaturated fatty acids, and increased levels of 4-hydroxynonenal (HNE), F(2)-isoprostanes, and F(4)-neuroprostanes are present in the brain in patients with AD. Acrolein, an alpha,beta-unsaturated aldehydic product of lipid peroxidation has been demonstrated to be approximately 100 times more reactive than HNE and is present in neurofibrillary tangles in the brain in AD. We recently demonstrated statistically significant elevated concentrations of extractable acrolein in the hippocampus/parahippocampal gyrus and amygdala in AD compared with age-matched control subjects. Concentrations of acrolein were two to five times those of HNE in the same samples. Treatment of hippocampal cultures with acrolein led to a time- and concentration-dependent decrease in cell survival as well as a concentration-dependent increase in intracellular calcium. In cortical neuron cultures, we now report that acrolein causes a concentration-dependent impairment of glutamate uptake and glucose transport in cortical neuron cultures. Treatment of cortical astrocyte cultures with acrolein led to the same pattern of impairment of glutamate uptake as observed in cortical neuron cultures. Collectively, these data demonstrate neurotoxicity mechanisms of arolein that might be important in the pathogenesis of neuron degeneration in AD. PMID- 11053773 TI - Metabolism of acetaldehyde to methyl and acetyl radicals: in vitro and in vivo electron paramagnetic resonance spin-trapping studies. AB - Acetaldehyde oxidation by enzymes and cellular fractions has been previously shown to produce radicals that have been characterized as superoxide anion, hydroxyl, and acetyl radicals. Here, we report that acetaldehyde metabolism by xanthine oxidase, submitochondrial particles and whole rats produces both the acetyl and the methyl radical, although only the latter was unambiguously identified in vivo. Electron paramagnetic resonance (EPR) characterization of both radicals was possible by the use of two spin traps, 5,5-dimethyl 1-pyrroline N-oxide (DMPO) and alpha-(4-pyridyl 1-oxide)-N-t-butylnitrone (POBN), and of acetaldehyde labeled with (13)C. The POBN-acetyl radical adduct proved to be unstable, but POBN was employed to monitor acetaldehyde metabolism by Sprague Dawley rats because previous studies have shown its usefulness for in vivo spin trapping. EPR analysis of the bile collected from treated and control rats showed the presence of the POBN-methyl and of an unidentified, biomolecule-derived, POBN adduct. Because decarbonylation of the acetyl radical is one of the routes for methyl radical formation from acetaldehyde, detection of the latter in bile provides strong evidence for the production of both radicals in vivo. The results may be relevant to understanding the toxic effects of acetaldehyde itself and of its more relevant biological precursor, ethanol. PMID- 11053774 TI - Cellular dissociation of NF-kappaB and inducible nitric oxide synthase in Helicobacter pylori infection. AB - The transcription factor nuclear factor kappaB (NF-kappaB) regulates the expression of inducible nitric oxide synthase (iNOS). We hypothesized that induction of iNOS in Helicobacter pylori gastritis may be due to NF-kappaB activation. Antral biopsy specimens from Helicobacter pylori-infected gastritis patients were collected before (n = 30) and after antimicrobial therapy to clear the infection (n = 22). Biopsies were assessed for NF-kappaB by immunohistochemistry (p65). The mRNA and protein of iNOS were localized by in situ RT-PCR and immunohistochemistry. Both of iNOS protein and mRNA were evident in stromal inflammatory cells, but absent in epithelia. Antimicrobial therapy resulted in a 73% reduction in iNOS levels (protein, p <.002). Nuclear staining for NF-kappaB p65 was evident in epithelial cells, especially in the neck region of gastric glands, and inflammatory cells. Treatment to clear H. pylori infection resulted in a 74% reduction in the epithelial staining for NF-kappaB p65 (p =.0001), whereas the lamina propria staining was unaltered. In conclusion, H. pylori infection activates NF-kappaB and iNOS expression. However, as the changes in NF-kappaB and iNOS with H. pylori clearance occurred in different cell types (epithelial vs. inflammatory), it appears that a NF-kappaB-dependent epithelial derived mediator may be responsible for the induction of iNOS expression. PMID- 11053775 TI - H(2)O(2)-induced egr-1, fra-1, and c-jun gene expression is mediated by tyrosine kinase in aortic smooth muscle cells. AB - Hydrogen peroxide (H(2)O(2)) has recently been shown to have a dual effect on cell growth by stimulating proliferation and triggering apoptosis. Apoptosis induced by H(2)O(2) is a direct consequence of oxidant injury, while the proliferative response to H(2)O(2) is thought to be a protective mechanism against oxidant injury. Signaling of the H(2)O(2)-induced proliferative effect has been proposed to occur via the activation of mitogen-activated protein kinase (MAPK) and increase in expression of transcription factors. In the present study, H(2)O(2)-induced mitogenic signaling in aortic smooth muscle cells (ASMC) was investigated with a specific focus on the roles of tyrosine kinase and tyrosine phosphatase in the regulation of the H(2)O(2)-stimulated egr-1, fra-1, and c-jun transcription. The results show that H(2)O(2)-induced increases in egr-1, fra-1, and c-jun mRNA levels, as measured by Northern blot analysis, are time and dose dependent with the peak of the response within 2 h. Tyrosine kinase inhibitors (genistein, amino-genistein, and tyrphostin 51) significantly attenuated H(2)O(2) induced expression of these genes and a tyrosine phosphatase inhibitor (perox vanadate) stimulated their expression. H(2)O(2) stimulated tyrosine kinase activities and caused protein tyrosine phosphorylation, which was blocked by tyrphostin 51. H(2)O(2) also caused tyrosine phosphorylation of platelet derived growth factor (PDGF) receptor. These data show that H(2)O(2) increases egr-1, fra 1, and c-jun mRNA levels in vascular smooth muscle cells, and the increase in expression of these genes is mediated by activation of tyrosine kinase. Our data also provide evidence that the H(2)O(2)-induced mitogenic response is, in part, mediated through the receptor tyrosine kinase, PDGF receptor. PMID- 11053776 TI - Oxidized low-density lipoprotein induces calcium influx in polymorphonuclear leukocytes. AB - Polymorphonuclear leukocytes (PMN) have been suggested to play a role in atherosclerosis, but intracellular signaling after stimulation with oxidized low density lipoprotein (LDL) is unknown. We investigated mechanistic aspects of oxidized LDL-induced superoxide production by human PMN, with special emphasis on intracellular Ca(2+) concentration ([Ca(2+)](i)). Oxidized LDL, but not native LDL, evoked an early but sustained increase in [Ca(2+)](i) and a delayed production of superoxide. The increase in [Ca(2+)](i) could be reduced by fucoidan and completely prevented by U73122, suggesting involvement of the scavenger receptor and coupling to the phospholipase C signal transduction pathway. Furthermore, we provide evidence that the increase in [Ca(2+)](i) partly results from protein kinase C-dependent Ca(2+) influx. The relevance of this Ca(2+) entry for oxidized LDL-stimulated effects is illustrated by the finding that superoxide production was markedly reduced in the absence of external Ca(2+). Finally, inhibition of phagocytosis by cytochalasin B abolished oxidized LDL-stimulated superoxide production without affecting, however, the Ca(2+) mobilization. These effects of oxidized LDL on [Ca(2+)](i) and on respiratory burst of PMN may underlie the occurrence of elevated levels of [Ca(2+)](i) of resting PMN in hypercholesterolemia and represent a mechanism by which PMN can amplify processes in the early phase of atherosclerosis. PMID- 11053777 TI - The involvement of the intracellular superoxide production system in hepatic ischemia-reperfusion injury. In vivo and in vitro experiments using transgenic mice manifesting excessive CuZn-SOD activity. AB - In vivo and in vitro studies were conducted using transgenic mice with 1.8-fold increased SOD activity in the cytoplasmic fraction compared to normal mice in order to evaluate the role of cytoplasmic superoxide dismutase (SOD) in hepatic ischemia-reperfusion injury. In the in vivo study, after inducing 15 min 70% partial hepatic ischemia followed by 45 min reperfusion, we determined the plasma levels of ALT, hyaluronic acid, and phosphatidylcholine hydroperoxide (PCOOH) as the membranous lipoperoxide of the hepatic tissue. In addition, in vitro ischemia reperfusion studies for cultured hepatocytes were conducted in an anaerobic chamber that could create a hypoxic or oxygen-rich environment in order to clarify the amelioration of reperfusion injuries in the SOD rich hepatocytes. High levels of ALT and PCOOH were found as a result of reperfusion in normal mice, while a suppression of the increase in these levels was noted in the transgenic mice. In both groups, the hyaluronic acid levels were not modified. These results suggest that intracellular superoxide production is involved in the mechanism of hepatic ischemia-reperfusion injury, and that an improvement of the ability to eliminate intracellular superoxide species can contribute to the prevention of reperfusion injury. PMID- 11053778 TI - Resuscitative effects of polynitroxylated alphaalpha-cross-linked hemoglobin following severe hemorrhage in the rat. AB - alphaalpha-Cross-linked hemoglobin (alphaalphaHb) is an example of a hemoglobin based oxygen carrier (HBOC) with significant cardiovascular activity. This may compromise the safety and efficacy of this HBOC by causing systemic hypertension and reducing blood flow to some organs. The present work is based on the hypothesis that incorporating antioxidant activity into an HBOC in the form of a covalently attached nitroxide may prevent these effects. We have tested this hypothesis by adding antioxidant activity to alphaalphaHb with 2,2,6,6 tetramethyl-piperidinyl-1-oxyl (Tempo) to create polynitroxylated alphaalphaHb (PN-alphaalphaHb). The new compound PN-alphaalphaHb acts as an antioxidant in our in vitro and in vivo assays. In this study urethane-anesthetized rats were hemorrhaged to a mean arterial pressure (MAP) of 35-40 mmHg and maintained for 30 min. Animals were resuscitated with solutions of (1) 10% PN-alphaalphaHb (43 mmHg), (2) 10% alphaalphaHb (43 mmHg), (3) 7.5% albumin (43 mmHg), (4) 300% Ringers lactate (RL), and (5) 0. 9% normal saline equal to the shed blood volume (SBV). Hemodynamics and regional blood circulation was measured at baseline, following hemorrhage, and at 30 and 60 min postresuscitation using a radioactive microsphere technique. Base deficit (BD) was measured at baseline, following hemorrhage, and at 60 min following resuscitative fluid infusion. Finally survival was determined as the time following resuscitation until secession of heart rhythm. Saline and 300% RL resuscitation did not improve BD, systemic hemodynamics, or regional blood circulation. PN-alphaalphaHb, alphaalphaHb, and albumin significantly improved these parameters, however, only PN-alphaalphaHb and alphaalphaHb improved survival. PN-alphaalphaHb was found to be less hypertensive than alphaalphaHb due to blunted increases in both cardiac output and systemic vascular resistance. This study demonstrates that, by using alphaalphaHb as a scaffold for polynitroxylation, improvement in vasoactivity and resuscitative efficacy may be possible. In conclusion, the addition of antioxidant activity in the form of polynitroxylation of a low molecular weight Hb (alphaalphaHb) may create a safe and efficacious resuscitative fluid. PMID- 11053779 TI - The reaction of superoxide radical with N-acetylcysteine. AB - The interaction of superoxide radicals with N-acetylcysteine (RSH) in an aqueous solution of pH 7 using the technique of steady state radiolysis has been investigated in this paper. The radiolytic yield of the products (G value) of RSH consumption and disulfide of N-acetylcysteine (RSSR) formation has been determined. The G value of the products is not dependent on the concentration of RSH (at the plateau of dilution curve) or on the inverse of the square root of the dose rate (dose rate)(-1/2), from which it is concluded that in this reaction there is no character of chain reaction. The disulfide of N-acetylcysteine is the only sulfur final product. Hydrogen peroxide is not a reaction product, and accordingly the reaction of O(2)(*-) with RSH does not proceed via hydrogen atom abstraction from RSH. A reaction mechanism is proposed, and an overall rate constant of 68 M(-1) s(-1) has been estimated. PMID- 11053780 TI - Products of the phospholipase A(2) pathway mediate the dihydrorhodamine fluorescence response evoked by endogenous and exogenous peroxynitrite in PC12 cells. AB - A short-term exposure of PC12 cells to tert-butylhydroperoxide promotes a rapid oxidation of dihydrorhodamine sensitive to nitric oxide synthase inhibitors and peroxynitrite scavengers. This response was not directly caused by peroxynitrite, but rather appeared to be mediated by peroxynitrite-dependent activation of phospholipase A(2). The following lines of evidence support this inference: (i) the peroxynitrite-dependent dihydrorhodamine fluorescence response was blunted by low concentrations of two structurally unrelated phospholipase A(2) inhibitors; (ii) under similar conditions, the phospholipase A(2) inhibitors prevented release of arachidonic acid; (iii) low levels of arachidonic acid restored the dihydrorhodamine fluorescence response in nitric oxide synthase- as well as phospholipase A(2)-inhibited cells; (iv) the dihydrorhodamine fluorescence response induced by authentic peroxynitrite was also blunted by phospholipase A(2) inhibitors and restored upon addition of reagent arachidonic acid. We conclude that endogenous, or exogenous, peroxynitrite does not directly oxidize dihydrorhodamine in intact cells. Rather, peroxynitrite appears to act as a signalling molecule promoting release of arachidonic acid, which in turn leads to formation of species causing the dihydrorhodamine fluorescence response. PMID- 11053781 TI - Alpha tocopherol supplementation decreases serum C-reactive protein and monocyte interleukin-6 levels in normal volunteers and type 2 diabetic patients. AB - Type 2 diabetic subjects have an increased propensity to premature atherosclerosis. Alpha tocopherol (AT), a potent antioxidant, has several anti atherogenic effects. There is scanty data on AT supplementation on inflammation in Type 2 diabetic subjects. The aim of the study was to test the effect of RRR AT supplementation (1200 IU/d) on plasma C-reactive protein (CRP) and interleukin 6 (IL-6) release from activated monocyte in Type 2 diabetic patients with and without macrovascular complications compared to matched controls. The volunteers comprised Type 2 diabetic subjects with macrovascular disease (DM2-MV, n = 23), Type 2 diabetic subjects without macrovascular complications (DM2, n = 24), and matched controls (C, n = 25). Plasma high sensitive CRP (Hs-CRP) and Monocyte IL 6 were assayed at baseline, following 3 months of supplementation and following a 2 month washout phase. DM2-MV subjects have elevated HsCRP and monocyte IL-6 compared to controls. AT supplementation significantly lowered levels of C reactive protein and monocyte interleukin-6 in all three groups. In conclusion, AT therapy decreases inflammation in diabetic patients and controls and could be an adjunctive therapy in the prevention of atherosclerosis. PMID- 11053782 TI - Electron paramagnetic resonance spectroscopy with N-methyl-D-glucamine dithiocarbamate iron complexes distinguishes nitric oxide and nitroxyl anion in a redox-dependent manner: applications in identifying nitrogen monoxide products from nitric oxide synthase. AB - Though a large number of studies indicate that nitric oxide synthase (NOS) is responsible for NO&z.rad; production in biological systems, controversy still remains concerning whether NOS directly produces NO&z.rad;. Schmidt et al. (PNAS 93:144492, 1996) proposed that NOS first synthesizes nitroxyl anion (NO(-)), which is then converted to NO&z.rad; by superoxide dismutase (SOD). With electron paramagnetic resonance spectroscopy using N-methyl-D-glucamine dithiocarbamate iron (Fe-MGD), we directly detected NO&z.rad; from purified NOS in the absence of SOD (Xia et al., PNAS 94:12705, 1997). We also showed that the requirement for SOD in the previous NO&z.rad; measurements appeared to be due to the high levels of exogenous superoxide production in their reaction system because of the presence of free FAD. However, it was recently questioned whether Fe-MGD can discriminate NO&z.rad; from NO(-) (Komarov et al., FRBM 28:739-742, 2000). In this study we examined the trapping specificity of different redox forms of Fe MGD. With Fe(2+)-MGD, NO&z.rad; generated characteristic triplet NO&z.rad;-Fe(2+) MGD signals (g = 2. 04, a(N) = 12.7 G), whereas NO(-) from Angeli's salt was EPR silent. Both NO&z.rad; and NO(-) gave rise to NO&z.rad;-Fe(2+)-MGD signals when Fe(3+)-MGD was used. Strong NO&z.rad; signals were measured from purified nNOS using the NO&z.rad; selective Fe(2+)-MGD and this was not affected by SOD. Thus, spin trapping with Fe-MGD can distinguish NO&z.rad; and NO(-) and this depends on the redox status of the iron. The detection of NO&z.rad; from purified NOS by Fe(2+)-MGD unambiguously reconfirms our previous report that NOS directly synthesizes NO&z.rad; but not NO(-). PMID- 11053783 TI - Febrile neutropenia: a necessary evil? PMID- 11053784 TI - Laboratory diagnosis of infections in febrile neutropenic or immunocompromised patients. AB - Laboratory diagnosis of infections in febrile neutropenic or immunocompromised patients is particularly challenging, and covers the most frequent clinical presentations such as bloodstream infections, lung, CNS and skin infections, as well as invasive fungal infections. Classic methods such as direct examination, culture and tests for susceptibility are being more complemented by molecular detection of microorganisms (PCR in particular) and antigen detection in various body fluids or tissues; two promising methods which offer the advantage of speed and high sensitivity even after starting antimicrobial therapy. PMID- 11053785 TI - Radiological and invasive diagnosis in the detection of pneumonia in fabrile neutropenia. AB - In the case of febrile neutropenia, a chest X-ray most often does not detect pneumonia and this may be related to the low sensitivity of this technique since high-resolution computed tomography (CT) has been shown to be suggestive of pneumonia in the majority of the cases when fever persists for more than 48 h. In such cases, early documentation of pneumonia is often possible. The pattern of pulmonary infiltrates on chest radiograph and CT scan can only be considered as indicative and confirmation of non-infectious or infectious processes is better achieved by bronchoalveolar lavage combined with transbronchial biopsy provided there is no contraindication. The results of the CT scan may also help to determine the site of sampling. PMID- 11053786 TI - Unusual presentations of infection in neutropenic patients. AB - Neutropenic patients may have unusual presentations of infection because of their inability to mount an adequate inflammatory response and their susceptibility to infection caused by less virulent organisms. About 60% of febrile episodes are associated with no other signs and symptoms and no infecting organism can be identified, yet most respond to antibacterial therapy. If not treated promptly, infection in neutropenic patients can progress rapidly. Unusual sites of infection include typhlitis, perirectal infections and atypical forms of cellulitis. PMID- 11053787 TI - New pathogens in neutropenic patients with cancer: an update for the new millennium. AB - As patients with malignant diseases are treated with increasingly potent agents it is likely that they will be subject to infection with an ever broadening array of microorganisms. As a result of the prompt institution of empirical antibiotics at the onset of fever in neutropenic patients, mortality has been reduced but new problems have emerged. First, there has been a shift in the type of infecting organisms responsible for bacteraemia in these patients from predominantly Gram negative organisms to Gram-positive cocci. Secondly, perhaps as a consequence of the effectiveness of antibiotics, there is increasing concern about infections with antibiotic-resistant organisms. As an example, viridans streptococci are becoming increasingly resistant to penicillin. Thirdly, organisms previously thought to be non pathogens or 'commensals' are now being reported as agents of serious invasive infections in neutropenic patients with cancer. This review will highlight these changes and discuss 'new' pathogens in these patients. PMID- 11053788 TI - Therapeutic guidelines for Pseudomonas aeruginosa infections. AB - Pseudomonas aeruginosa nowadays is encountered among the leading pathogen in (i) ICU pneumonia; (ii) nosocomial bacteremia and AIDS primary bacteremia; (iii) iv drug users endocarditis; (iv) exacerbations of cystis fibrosis; (v) malignant external otitis and 'swimmers's ear', and (vi) contact lenses keratitis and traumatic endophthalmitis. The most vulnerable nosocomial hosts are the neutropenics and the mechanically ventilated patients in whom mortality rate exceeds 30%. Virulence of P. aeruginosa is attributed to the elaboration of various enzymes and toxins. There is also worldwide emergence of multiresistant phenotypes to antipseudomonal antibiotics. Therapeutic guidelines should therefore be based on (i) continuous resistance surveillance; (ii) in vitro synergistic interactions of antibacterial agents; (iii) pharmacodynamic properties of antibiotics interpreted by optimal dosing and appropriate frequency of administration; and (iv) current information on the necessity for combination therapy using an aminoglycoside. PMID- 11053789 TI - Risk factors assessment in fabrile neutropenia. AB - It is now established that febrile neutropenic cancer patients constitute a heterogeneous population with a variable risk for serious medical complication development. Optimal patient management should take that risk into account by replacing, for instance, the classical, in-hospital administered, broad-spectrum intravenous antibiotics by newer therapeutic approaches including oral and/or outpatient therapeutic strategies for the 'low-risk' patients. The development of such approaches which have been shown safe and feasible, implies the existence of universally accepted, validated and reliable clinical prediction rules for the identification of these low-risk patients. Some prognostic factors predicting the response to the empiric treatment, the development of a bacteremia, and the final outcome of a febrile neutropenic episode have been established (such as duration and profoundness of neutropenia, acute leukemia, administration of chemotherapy for treatment of relapse, high temperature, shock and/or chills, inpatient status at fever onset) and some models combining them have already been proposed, firstly by Talcott and coworkers and more recently by the Multinational Association for Supportive Care in Cancer (MASCC) study section on infections. The sensitivity of these rules as a selection tool for identifying patients at low-risk of complication, however, needs to be improved and we have to assess their clinical usefulness, safety and/or reproducibility better in order to allow a more adequate choice between the therapeutic strategies, to continue to improve patients quality of life and to optimize the cost-effectiveness of the treatments. PMID- 11053790 TI - Prediction of neutropenia. AB - Newer treatment strategies for the management of febrile neutropenic patients are being developed. These include: (a) hospital based oral therapy; (b) early discharge after initial stabilization in-hospital, and (c) out-patient therapy. All strategies are likely to be more successful in patients with short-lived neutropenia (< or = 7 days) than in those with more prolonged neutropenia. Although risk-prediction rules and clinical criteria can help clinicians identify 'low-risk' neutropenic patients, the overall ability of clinicians to accurately predict the subsequent duration of neutropenia once a neutropenic patient becomes febrile needs to be improved upon considerably. One attempt to do so is the survey being conducted by the Infection Study Section of the Multinational Association of Supportive Care in Cancer (MASCC). Development of an accurate rule for the prediction of neutropenia will enable more appropriate, risk-based therapy to be administered to febrile neutropenic patients, and will represent a significant advance in the management of such patients. PMID- 11053791 TI - Colony stimulating factors in patients with fever and neutropenia. AB - Colony stimulating factors (CSFs), are essential for the regulation of the hematopoietic system and were developed by the pharmaceutical biotechnology industry in the early 1990s for the prevention of serious neutropenic complication after myelosuppressive chemotherapy. Both G-CSF and GM-CSF consistently lead to an increase in circulating white blood cells and a reduction in the incidence of fever and neutropenia after myelosuppressive chemotherapy in patients with solid tumors, hematologic malignancies, and those undergoing stem cell transplantation. Their role in improving response rates to chemotherapy and overall survival is less clear. PMID- 11053792 TI - Antibacterial prophylaxis. AB - Bacterial infections remain an important cause of morbidity and mortality in neutropenic patients. A number of prophylactic strategies have been used in order to reduce the risk of infection during severe granulocytopenia. The measures that have been investigated include isolation of the patient, granulocyte transfusion, active or passive immunisation, acceleration of granulocyte recovery and prophylactic use of antibacterial agents. However, many of these approaches have fallen out of favour, mostly because of concerns about the long lasting efficacy. This paper focuses on the available prophylactic strategies, with emphasis on the use of antibacterial agents. PMID- 11053793 TI - Febrile neutropenia: antifungal prophylaxis. AB - Despite being widely practised there is little or no evidence to support the use of antifungal prophylaxis in febrile nuetropenic patients. With the greater recognition of risk factor, the increasing readiness to use imaging techniques such as the high-resolution CT scan and new methods for detecting fungi such as Aspergillus galactomannan on the one hand and the widening choice of drugs on the other hand, it may be possible to move away from prophylaxis towards preemptive treatment when the prevalence of fungal disease is low. However, the case still has to be proven convincingly by conducting formal clinical trials. PMID- 11053794 TI - Empirical treatment of sepsis in neutropenic patients. AB - Febrile neutropenia remains a major cause of morbidity in cancer patients receiving chemotherapy. Although the mortality associated with febrile neutropenia has dramatically decreased over the last three decades, the overall death rate during and immediately after an episode of febrile neutropenia can be as high as 10% with half of the patients dying directly as a result of the infection itself. A series of developments has led to this marked reduction in mortality. Among them, a pivotal role has been played by the concept of hospital based empirical therapy with broad-spectrum combinations of antibiotics, aimed primarily against Gram-negative organisms, namely Pseudomonas aeruginosa PMID- 11053795 TI - Management of fungal infections in neutropenic patients: more doubts than certainties? AB - An evidence based approach to prophylaxis and therapy of invasive fungal infections depends on the knowledge of epidemiology and of risk factors for these infections, as well as on the appreciation of merits and limitation of the available clinical trials. A progressive increase in the incidence of systemic fungal infections, most often caused by Candida and Aspergillus, in patients with cancer and neutropenia has been observed in recent years. This increase of systemic fungal infections recognizes a multifactorial origin, including host defense impairment and type of underlying disease. The various combinations of these different risk factors make the patients affected by systemic fungal infections a non-homogeneous population and, therefore, the transferability of the results of many clinical trials from one population to another is difficult. Clinical trials on prophylaxis and treatment of systemic fungal infections moreover have many limitations: they are often of small size, are frequently non comparative, enrol population at different risk for infection, use different criteria to define success or failure of therapy. These limitations make the interpretation of the trial results difficult. As randomised clinical trials and metanalysis are considered the most valuable sources of information on new treatments, it dearly appears that the mentioned difficulties in interpreting available data from the literature may expose patients to an increased risk of receiving an inappropriate or non-optimal treatment. Better designed studies are needed to clarify the many controversial questions in antifungal prophylaxis and therapy. PMID- 11053796 TI - Modifications of therapy. AB - Rates of response to empirical therapy in the neutropenic patient with fever, range between 40 and 90%. Modifications of therapy are needed in patients who do not respond but may also be considered in patients who respond. In patients with unexplained fever and rapid defervescence, switch to oral therapy is an acceptable option and there is no need to continue the regimen until neutrophil recovery. Neutropenic patients with persistent undefined fever and those with progressive pneumonia benefit from the addition of antifungals while the empiric addition of a glycopeptide is unlikely to be effective. In patients with gram negative bacterial infection initially treated with monotherapy, response may be increased after the addition of an aminoglycoside. In cases of a defined etiology, the institution of narrow-spectrum antimicrobials in a persistent neutropenic patient carries a substantial risk for superinfection and is not generally recommended. Improved diagnostic tools and sensitive clinical risk assessment methods will allow selecting and targeting therapy modifications better. PMID- 11053797 TI - Standard antifungal therapy in neutropenic patients. AB - Fungal infections are life-threatening complications in patients with prolonged neutropenia. Amphotericin B, which is fungicidal and has a broad spectrum of antifungal activity, remains the current gold standard agent. However, because of its low therapeutic index, new lipid formulations of amphotericin B have been developed with better tolerance and less toxicity. The use of 5-fluorocytosine is waning because of the medullar toxicity of this agent. Fluconazole and itraconazole are both fungistatic and are more convenient for the treatment of fungal infections in haemodynamically stable patients. PMID- 11053798 TI - New antifungal agents and preparations. AB - In neutropenic patients amphotericin B remains the drug of choice for the treatment of systemic fungal infections. On the basis of a superior efficacy in combination with a lower toxicity, the triazoles have superseded the older azoles. Regularly, amphotericin B and a triazole are used simultaneously without any evidence from clinical trials that such a strategy is safe and efficacious. Liposomal preparation, lipid complex or colloidal dispersion of amphotericin B have been produced successfully to reduce toxicity. However, there is only one small randomised study that hints at the superiority of liposomal amphotericin B over amphotericin B deoxycholate. Promising new agents like candins, sordarins, high dose oral terbinafine, the third generation azoles, and liposomal nystatin are under development. The first phase II study on voriconazole in the treatment of pulmonary aspergillosis has produced encouraging results. The major promise of the new candins lies in the activity against Candida species, including those resistant to the azoles and polyenes, and in a mechanism of action totally different from the established antifungals. Cytokines and colony stimulating factors are theoretically very promising but there are no clinical studies that warrant routine use. PMID- 11053799 TI - Concepts in design of comparative clinical trials of antifungal therapy in neutropenic patients. AB - Fundamental to the successful implementation of antifungal compounds in neutropenic patients is the appropriate design of comparative clinical trials investigating their safety and efficacy. The key elements of comparative clinical trial design include issues of enrollment, stratification, randomization, blinding, administration of study drugs, monitoring of drug toxicity, definitions, and key statistical elements of end points, sample size, and tools for data analysis. The initial selection of compounds and the timing of initiation of antifungal therapy in comparative clinical trials are predicated to a large degree on the in vitro and in vivo activities, plasma pharmacokinetics, profiles of safety and toxicity of the study drugs. Phase I and II studies have a critical role in designing comparative clinical trials of antifungal therapy by providing data on safety, tolerance, and plasma pharmacokinetics of the investigational agent. As new antifungal agents are developed in response to the challenge of invasive fungal infections in immunocompromised patients with cancer, thoughtfully designed and carefully implemented clinical trials will be essential in determining the future utility of these promising compounds. PMID- 11053800 TI - Antiviral therapy of herpes simplex. AB - Herpes simplex virus (HSV) infections in immunocompromised patients are more severe and invasive than in non-immunocompromised patients. They are characterised by prolonged viral shedding and a tendency to heal more slowly. In addition, resistant viruses are exclusively isolated in immunocompromised patients, requiring other drugs with distinct mechanisms of action. The reference compound for the treatment of HSV infections is acyclovir (ACV) that selectively inhibits HSV DNA replication with low host-cell toxicity. Recently, two molecules, valaciclovir (VACV), the L-valyl ester of ACV and famciclovir (FCV), the diacetyl ester of 6-deoxy-penciclovir (PCV), another potent nucleoside analogue, were developed showing an increased oral bioavailability compared to the original compounds. Foscavir (PFA) and more recently cidofovir (CDV) are drugs that do not need the viral thymidine kinase (TK) to be activated and therefore are the appropriate candidates for the treatment of resistant viruses emerging under acyclovir or penciclovir. PMID- 11053801 TI - Prevention and management of mucositis in patients with cancer. AB - This review summarises the large number of locally and systemically applied preventive and therapeutic interventions of mucositis in patients with cancer. The need for further elucidation of the pathophysiology and for optimisation of trial methodology is emphasised. Data from trials in animal models and preliminary data in patients indicate that cytokines such as interleukin-1, interleukin-11, TGF-beta 3 and keratinocyte growth factor could reduce the incidence of mucositis. Other potentially useful agents are the angiogenesis inhibiting drug thalidomide, the cytoprotector amifostine and the pineal hormone melatonin. PMID- 11053802 TI - Intravenous access: related problems in oncology. AB - This article reviews the latest developments in the literature concerning two types of catheter widely used in oncology: Hickman central venous catheters (CVCs) and totally implanted venous ports. It now seems possible to diagnose catheter-related bloodstream infections (CRBI) without removing the device; recent studies showed that paired quantitative blood cultures are the most reliable and convenient method for diagnosis of CRBI. Can CRBI be cured and treated without device removal? A decision-tree/flow-chart resulting from analysis of international data by French International Experts is presented. Futures trends are reviewed: new techniques for prevention, including catheters impregnated with antimicrobials and, antiseptic hubs; flushing and antibiotic lock techniques that can be used both for prevention and treatment. PMID- 11053803 TI - Out-patient management of febrile neutropenia. AB - Fever and neutropenia commonly complicated cytotoxic cancer therapy. Although standard therapy is empirical in-patient broad-spectrum intravenous antibiotic therapy until fever and neutropenia resolve, different treatments based on patient risk have been reported over the last decade. The data identifying low risk patients, treatment strategies for low risk patients, and additional information required before recommending out-patient management of fever and neutropenia widely are described. PMID- 11053804 TI - Febrile neutropenia in children. AB - Fever is frequent in neutropenic patients and often related to infection. Two major concepts, have contributed to the marked mortality decrease of those patients by the end of the 1960s: firstly, the duration and severity of neutropenia were the most important variables linked to infection and secondly, prompt administration of broad-spectrum antimicrobials empirically, was life saving. At the same time it was universally admitted that a careful daily examination of all portals of entry for micro-organisms was mandatory and that laboratory and imaging investigations were needed at regular intervals, keeping constantly in mind the individual type and stage of immunosuppression. Through many studies, paediatricians contributed markedly in standardisation of management of febrile neutropenic patients. Neutropenic patients are not equally prone to infections, partly due to the underlying cancer, chemotherapy and co morbidity factors. Neutropenic children are not only vulnerable to bacteria, fungi and viruses commonly encountered in adults, but also to common viruses and bacteria. Very few studies included a viral work-up. Epidemiological new trends are observed: Gram-positive bacteria and fungi are on the rise. Simplifying and shortening antibiotic regimens were made possible because new potent antibiotics were launched. Since the mid-1980s, many paediatric centres commonly discharge patients before complete bone marrow recovery, provided that patients meet certain low-risk criteria and do not exhibit any clinical or biological evidence of bacterial infection. However, a few prospective randomised studies have been conducted for assessing the safety of early antibiotics discontinuation and safe early discharge. The choice of oral agents up to now was complicated by the reluctance using fluoroquinolones in children. New challenges are numerous in terms of diagnostic tools, detection of epidemiological trends and emerging pathogens, identification and control of nosocomial threats including drug resistance, assessment of the real impact of prophylaxis, evaluation of new agents, the need for more accurate risk scoring systems, outpatient management and the necessity for an optimal use of resources. PMID- 11053805 TI - Febrile neutropenia in allogeneic transplantation. AB - Infections post allogeneic bone marrow transplant (BMT) are a major problem. Post BMT, three periods with infectious complications are discerned: pre-engraftment and early recovery, mid recovery and late recovery. In the first period mucosal damage and neutropenia are the major host defence deficits. Bacterial infections with Gram-positive and Gram-negative organisms, and fungal infections are seen in this period. In the mid recovery phase graft versus host and its treatment contribute to diminished host defences. Viral infections and fungal infections predominate. In the late recovery phase chronic graft versus host reaction impairs the monocyte macrophage function and CD4 counts are low. In this phase patients are at risk for infections with encapsulated bacteria, fungi, Pneumocystis carinii and Toxoplasma. Strategies for the management of febrile neutropenia are similar to those in 'high risk' neutropenic patients: immediate broad spectrum I.V. antibiotics (3rd or 4th generation cephalosporin+/ aminoglycoside or carbapenem) and early amphothericin B (lipid formulation) if fever persists beyond 5 days despite adequate I.V. antibiotics. Cytomegalovirus (CMV) prophylaxis or better preemptive therapy guided by viraemia is accepted practice. Prevention of infection measures, antimicrobial, antifungal and viral prophylaxis are generally accepted strategies but would differ from center to center. The post transplant infection history will change with different transplant techniques and evolving prophylactic and preemptive treatments. PMID- 11053806 TI - Medical economic considerations of supportive cancer care. AB - In an era of increasing cost containment in health care, it has become extremely important to allocate resources as efficiently as possible. This implies that formal health economic considerations are taken into account. One of increasing importance is the field of economic appraisal, which allows to quantify the value for money of medical interventions. This research is extremely useful in the field of supportive care of cancer patients. Economic considerations, social, political and ethical issues will also have to be addressed. The sum of these activities will enable us to make better choices in health care and ensure that sufficient resources are allocated to supportive cancer treatment. Cancer patients have physical, social, spiritual and emotional needs. They may suffer from severe physical symptoms, from social isolation, spiritual abandonment, and emotions such as sadness and anxiety, or feelings of deception, helplessness, anger and guilt. In some of them, the disease is rapidly progressing and ultimately they die. Their demanding care evokes intense feelings in health care providers, all the more since these incurable patients represent a challenge, which could be condensed under the heading "the challenge of medical omnipotence". The complexity of the matter, the interdisciplinary approach and the emergence of an increasingly cost conscience health care environment put supportive cancer care in a difficult realm. PMID- 11053807 TI - Multinational cooperation in trials and guidelines dealing with febrile neutropenia. AB - Multinational cooperation has lead to many of the advances in the therapy of cancer patients with febrile neutropenia over the last two decades and will likely continue to do so. Some of the groups and societies that have contributed to this success include the European Organization for Research and Treatment of Cancer (EORTC) Antimicrobial Group, The Immunocompromised Host Society (ICHS), the Multinational Association for Supportive Care in Cancer (MASCC) infection subcommittee and the International Disease Society of America (IDSA). Some of these contributions are described in this manuscript. Without this approach the data from single institutions may not have been so well accepted in centers all over the world. Multinational cooperation in this field along with better antimicrobials have contributed significantly to the drop in mortality in 'high risk patients' from >50% in the 1950s and 1960s to 5-10% noted in most recently published studies. PMID- 11053808 TI - The role of airway stents in the management of pediatric tracheal, carinal, and bronchial disease. AB - OBJECTIVE: A variety of stents are available to aid in the management of complex tracheal, carinal and bronchial stenoses. We reviewed our multi-institutional experience with airway stents in children. METHODS: Thirty-three children (age, 13 days-18 years) from four institutions have had a total of 40 stents placed to aid in the management of complex airway stenoses. Three stent types were utilized: 29 silastic stents, five expandable metal stents and six customized carinal stents (four patients had two stents and one patient had four stents). Thirty children had tracheal stents, six children had bronchial stents, and two infants had carinal stents (three children had stenting of more than one area and two had stenting of all three locations). Twenty-eight patients (age, 5 months-18 years; mean, 8.06 years; SEM, 1.13 years) had stents placed after a variety of airway reconstructive procedures. Four underwent stenting in a non-operative setting and one as preoperative stabilization. RESULTS: Twenty-seven patients survived. One patient died early due to bleeding. Five patients died late: two due to bleeding, one from mediastinitis, and two patients with functional airways died late from unrelated problems. Complications are related to stent type and location. Carinal stents can migrate; several techniques are available to help manage this problem. Wire stents are essentially non-removable requiring periodic dilation. Silastic stents stimulate granulation tissue formation requiring periodic bronchoscopic removal. CONCLUSION: Tracheal stenting can aid in the management of pediatric airway problems. Complications are common, but can be managed with appropriate intervention. PMID- 11053809 TI - Factors affecting long-term survival after en-bloc resection of lung cancer invading the chest wall. AB - OBJECTIVE: Several reports emphasize the importance of en-bloc resection as the optimal surgical treatment of lung cancer with chest wall invasion. We investigated possible factors which could affect long-term survival following radical resection of these tumors. METHODS: Between 1981 and 1998, 100 patients (90 male; ten female), with a median age of 60 years (36-84), underwent radical en-bloc resection of non-small cell lung cancer (NSCLC) with chest wall involvement. Patients with superior sulcus tumors invading the thoracic inlet were excluded from this series. There were 43 squamous and 57 non-squamous tumors. The median number of resected ribs was three (1-5). Lung resection included 73 lobectomies, two bilobectomies, 18 pneumonectomies and seven segmentectomies. Chest wall resection also extended to the sternum in one patient, the transverse process in one, the costotransverse foramen and hemivertebrae in two. All patients had a complete resection. Sixty-three patients received postoperative radiotherapy and 12 received chemotherapy. Histological data, including differentiation and depth of chest wall invasion, were carefully reviewed. The effect of various factors on survival were studied. RESULTS: There were four in-hospital deaths. Lymph node involvement was negative on surgical specimens in 65 patients, and 28 patients had positive N1 nodes; the final histology revealed seven N2 diseases. Chest wall invasion was limited to the parietal pleura in 29 patients and included intercostal muscles, bones and extrathoracic muscles in 67, 24 and seven cases, respectively. The overall 2-year survival rate was 41%. The 5-year survival for patients with N0, N1 and N2 disease was 22, 9 and 0%, respectively. A local recurrence occurred in 13 patients, with four having a new resection and 45 patients developing systemic metastases. The nodal status (N0-1 vs. N2; P=0. 026) and the number of resected ribs(<2 vs. >2; P=0.03) were survival predictors in univariate analysis. By multivariate analysis, the two independent factors affecting long-term survival were the histological differentiation (well vs. poorly differentiated; P=0. 01) and the depth of chest wall invasion (parietal pleura vs. others; P=0.024). CONCLUSIONS: Histological differentiation and depth of chest wall involvement were the main factors affecting long-term survival in this series. The role of induction chemotherapy for tumors with poor prognosis should be investigated. PMID- 11053810 TI - Analysis of risk factors in bronchopleural fistula after pulmonary resection for primary lung cancer. AB - OBJECTIVES: Despite the advances in surgical technology, bronchopleural fistulas (BPFs) still occur and are often fatal. We studied the risk factors for BPF formation after lung cancer operation to clarify the indication of preventive bronchial stump coverage. In addition, the reliability of our methods of bronchial closure was evaluated. METHODS: We reviewed 557 consecutive bronchial stumps, corresponding to 547 patients without any coverage in pulmonary resection for lung cancer between 1989 and 1998. According to nine variables, stumps that made dehiscence were compared with uneventful ones using contingency table analysis. The incidence of BPFs according to each method of bronchial closure was calculated. RESULTS: BPFs developed in ten patients (1.8%). Compared with the lobar bronchus (LB), the main bronchus (MB; P<0.01; odds ratio, 23.0) and the intermediate bronchus (IB; P=0.03; odds ratio, 10.7) carried a high risk. Previous ipsilateral thoracotomy (P<0.01; odds ratio, 37.9) and preoperative chemotherapy and/or radiotherapy (P=0.02; odds ratio, 13.2) increased the risk. The incidence of BPFs with manual suture, stapling devices only, reinforcement suture at the distal side of staplers, or reinforcement suture at the proximal side of staplers was 1.8, 5.0, 1.9 and 1.0%, respectively. CONCLUSIONS: The main and intermediate bronchial stumps, and the stumps in cases with previous ipsilateral thoracotomy or receiving induction therapy are prone to BPFs. Preventive coverage should be considered for these stumps. Our methods for reinforcement of stapled stumps are thought to be reliable. PMID- 11053811 TI - Lung cancer following previous extrapulmonary malignancy. AB - OBJECTIVE: Having demonstrated a poor prognosis of operable lung cancer in patients with previous head and neck malignancies, we intended to evaluate prognosis of lung cancer in patients with a history of extrapulmonary and extracervical malignancies. METHODS: The population of this study included 55 patients; these were 40 males and 15 females, with a mean age 64.4+/-8.6 years. The previous malignancy was considered tobacco-induced in 15 patients (kidney, two; bladder, ten; esophagus, three), hormone-dependant in 18 (breast, six; female genital, eight; prostate, four), and miscellaneous in 22 (leukemia, four; skin, seven; colon, 11). Following complete resection, 25 patients were classified stage I, 13 were stage II, and 17 were stage IIIA. RESULTS: There were two early perioperative deaths (3.6%), and three during the second month owing to cardiovascular complications. At the conclusion of the study (July 1st, 1997), 32 further patients had died (58.2%): 25 had progression of lung cancer, one had progression of previous malignancy, and six were without evidence of disease. Five-year survival (Kaplan-Meier) was estimated 47+/-10.2% in stage I (median 44 months), 30.8+/-15.6% in stage II (median 26 months), and 16. 7+/-9.9% in stage IIIA (median 17 months). When excluding five early perioperative deaths, 5-year survival was 51.1+/-10.6% in stage I (median 93 months), 33.3+/-16.7% in stage II (median 36.5 months), and 19.0+/-11.2% in stage IIIA (median 20.5 months). Comparing the three groups defined according to location of previous malignancy, there was no significant difference neither in stage distribution (chi(2)=1.326; P=0.857), nor in 5-year survival estimates: 38.9+/-12. 9% (median 27 months) after tobacco-induced malignancies, 38.9+/-11. 5% (median 24 months) following hormone-dependant malignancies, and 28.4+/-10.2% (median 28 months) following miscellaneous cancers (chi(2)=0.059; P=0.9707). CONCLUSIONS: In opposition to data collected in patients with previous head and neck cancer, survival estimates according to stage were contained within the universally accepted range no high risk group has been identified. Resection of lung cancer with curative intent is a fair option in patients with previous extrapulmonary malignancy. PMID- 11053812 TI - Iterative surgical resections for local recurrent and second primary bronchogenic carcinoma. AB - OBJECTIVE: To report our experience with repeated pulmonary resection in patients with local recurrent and second primary bronchogenic carcinoma, to assess operative mortality and late outcome. METHODS: The medical records of all patients who underwent a second lung resection for local recurrent and second primary bronchogenic carcinoma from 1978 through 1998 were reviewed. RESULTS: There were 27 patients. They constituted 2.5% of 1059 patients who had undergone lung resection for bronchogenic carcinoma in the same period. Twelve patients (1.1%) (group 1) had a local recurrence that developed at a median interval of 24 months (range 4-83). The first pulmonary resection was lobectomy in ten patients and segmentectomy in two. The second operation consisted of completion pneumonectomy in ten cases, completion lobectomy in one and wedge resection of the right lower lobe after a right upper lobectomy in one. The other 15 patients (1.4%) (group 2) had a new primary lung cancer that developed at a median interval of 45 months (range 21-188). The first pulmonary resection was lobectomy in 12 patients, bilobectomy in one and pneumonectomy in two. The second pulmonary resection was controlateral lobectomy in seven patients, controlateral sleeve lobectomy in two, controlateral pneumonectomy in 1, controlateral wedge resection in four and completion pneumonectomy in one. Overall hospital mortality was 7.4%, including one intraoperative and one postoperative death in group 1 and 2, respectively. Five-year survival after the second operation was 15.5 and 43% with a median survival of 26 and 49 months in groups 1 and 2, respectively (P=ns). CONCLUSIONS: Long-term results justify complete work-up of patients with local recurrent and second primary bronchogenic carcinoma. Treatment should be surgical, if there is no evidence of distant metastasis and the patients are in good health. Early detection of second lesions is possible with an aggressive follow-up conducted maximally at 4 months intervals for the first 2 years and 6 months intervals thereafter throughout life. PMID- 11053813 TI - Surgical repair of the pulmonary trunk aneurysm. AB - OBJECTIVE: Aneurysm formation of the pulmonary trunk is rare and there is controversy about optimal treatment for this disease. The aim of this article is to report four patients with pulmonary trunk aneurysm which were managed by surgical repair. MATERIALS AND METHODS: From 1986 to 1997, we performed surgical repair for pulmonary trunk aneurysm in four patients. There was one male and three female patients with a mean age of 63.3 years (range: 54-78 years). Concomitant diseases were cardiac valvular disease in four patients, thoracic aortic dissection in two, atherosclerotic abdominal aortic aneurysm in two, and coronary artery disease in one. All patients were in New York Heart Association functional class III preoperatively. Surgical procedures for the pulmonary trunk aneurysm included Dacron graft replacement in two patients and aneurysmorrhaphy in two. Associated procedures were cardiac valvular operation in three patients with four lesions and right ventricular outflow tract reconstruction (RVOTR) in one. RESULTS: There were no operative mortalities and no late deaths with a mean follow-up period of 6.6 years (range: 2.4-10.0 years). One female patient developed recurrent pulmonary trunk aneurysm 9.5 years after aneurysmorrhaphy, and underwent a second operation where Dacron graft replacement of the aneurysm including pulmonary valve replacement was performed successfully. All patients are now leading normal lives. CONCLUSIONS: Surgical management should be considered for large aneurysm of the pulmonary trunk regardless of its etiology and underlying disease to prevent possible rupture with fatal result if the patient has an acceptably low operative risk. PMID- 11053814 TI - Considerations in biventricular repair after the Norwood procedure. AB - OBJECTIVE: The Norwood procedure can be applicable as a first stage palliation in children who can eventually undergo a biventricular repair. Although usual management of these patients is a primary neonatal repair, in selected patients staged approach with a Norwood procedure in the neonatal period followed by a Rastelli procedure in the infancy for conversion to two-ventricle physiology has been used alternatively. METHODS: We report our experiences on two infants who underwent a previous palliation with the Norwood procedure for lesions other than hypoplastic left heart syndrome and converted to two-ventricle physiology by the use of a Rastelli-type procedure. This report examines considerations in biventricular repair after the Norwood procedure especially need for ventricular septal defect enlargement and approach to placement of the right ventricle to pulmonary artery conduit. RESULTS: Both of the infants who underwent staged approach with an initial Norwood procedure for lesions other than hypoplastic left heart syndrome survived the operations and were clinically well at mid-term follow-up. CONCLUSION: In selected patients, the staged approach is an alternative in management of malformations other than hypoplastic left heart syndrome which share the important physiologic features of aortic outlet obstruction and ductal dependency of systemic circulation. We recommend routine enlargement of ventricular septal defect and proper positioning of the conduit at the time of subsequent biventricular repair. PMID- 11053815 TI - Aortic arch repair using hypothermic circulatory arrest technique associated with pharmacological brain protection. AB - OBJECTIVE: Hypothermic circulatory arrest is a standard procedure for the treatment of aortic arch. However, there is a time limit for this procedure. There is now an urgent need to develop prophylactic measures to extend the time limit. We have used a pharmacological mixture of thiopental, nicardipine and mannitol for all patients undergoing circulatory arrest since 1991 to extend the safe limit. The purpose of this study was to analyze the neurological complications demonstrated by these patients and to evaluate the brain-protective effects of our measure. METHODS: The clinical records of 75 consecutive patients undergoing an aortic arch repair using a hypothermic circulatory arrest technique during the past 8 years were retrospectively reviewed. Systemic cooling was continued until a total disappearance of EEG activity. Prior to circulatory arrest, 15 or 30 mg/kg of thiopental, 20 mg of nicardipine and 300 ml of mannitol were infused into the venous reservoir of a cardiopulmonary bypass circuit. Graft replacement was performed in all patients and the extent of replacement was a total aortic arch in 43 patients, a distal aortic arch in 17, a hemiarch in 13 and a distal aortic arch and a total descending aorta in two. RESULTS: The duration of circulatory arrest ranged from 16 to 80 min (mean 41.5 min), and it exceeded 45 min in 37 patients. Operative mortality was 10.7% and two patients died of stroke. Three patients had permanent and three other patients had transient neural deficits. The incidence of stroke was 8.0% as a whole, and no correlation between the incidence of neurological complications and the duration of circulatory arrest was found. A multivariate analysis showed that the duration of circulatory arrest was determined as a predictor of neither operative mortality nor postoperative stroke. CONCLUSIONS: The findings of the present study suggest that our pharmacological brain protection appears to be effective for safely extending hypothermic circulatory arrest. PMID- 11053816 TI - Mid-term results of partial left ventriculectomy in end-stage heart disease. AB - OBJECTIVE: Immediate and mid-term effectiveness of partial left ventriculectomy (PLV) is assessed in 27 idiopathic dilated cardiomyopathy patients. METHODS: All patients were in New York Heart Association (NYHA) class III (17) or IV (ten). The average left ventricular ejection fraction (LVEF) was 19+/-4% by MUGA, and 23+/-4% by digital echocardiography. The mean end-systolic volume (LVESV) was 259+/-66 ml and the mean end-diastolic volume (LVEDV) was 342+/-83 ml. Mitral valve replacement was a routine part of the procedure. RESULTS: Operative mortality was 18.5%, a LVEDP>25 mmHg, left atrial diameter>55 mm, pulmonary artery systolic pressure>40 mmHg, congestive hepatomegaly and NYHA class IV being the mortality predictors. Three-year Kaplan-Meier survival was 64+/-10%, including operative mortality; freedom from congestive heart failure was 65+/ 11%. Functional status improved from 3.2+/-0.4 to 1.5+/-0.6 (P=0.0003). The mean LVEF was dramatically increased after PLV (to 40+/-4%, P=0.0001); LVESV was decreased to 90+/-30 ml (P<0.0001) and LVEDV to 160+/-49ml (P<0.0001). This improvement was sustained during the first 30 months. CONCLUSIONS: PLV is a reasonable approach for end-stage patients, providing sustained dramatic changes in ventricular geometry and functional capacity, especially in the absence of compromised right and diastolic left heart functions. Routine replacement of the mitral valve allows a more liberal ventriculectomy and eliminates mitral regurgitation, and this may help minimize ventricular distention. PMID- 11053817 TI - Assessment of changes in general health status using the short-form 36 questionnaire 1 year following coronary artery bypass grafting. AB - OBJECTIVE: The problem addressed in the study was to gain a greater understanding of the health benefits of coronary artery bypass grafting (CABG). The purpose of the study was to assess general health status, using the short-form (SF)-36 questionnaire, approximately 12 months following CABG, and to document any associations between pre-operative health status, level of social support, coronary artery disease (CAD) risk factors, CAD symptom severity and post operative health status. METHODS: The study was prospective and observational in design and included assessments at two time points, namely pre-operatively in a hospital outpatient department (1995-1996) and post-operatively at home (1996 1997). Two hundred and fourteen patients awaiting elective CABG were recruited a month before the expected date of operation. Pre-operative assessment included: (1), severity of symptoms; (2), CAD risk factors; (3), SF-36 questionnaire; and (4), social activities questionnaire. Post-operative assessment measured health status using the SF-36 instrument (mean, 16.4 months). Correlation and multiple linear regression analyses were used to identify factors associated with improved health status following CABG. RESULTS: Two hundred and fourteen patients were assessed pre-operatively and underwent CABG. There was a 4.8% 30-day mortality rate, and 183 patients were followed for a mean of 16.4 months after CABG. SF-36 scores following CABG were improved across all of the eight domains (P<0.001). A higher social network score and higher pre-operative health status were associated with improved health status. Patients with lower health levels (SF-36 scores) prior to CABG were less likely to gain improvement in health (SF-36 scores) following CABG. Lower SF-36 scores following operation were influenced by the presence of diabetes mellitus, cigarette smoking, younger age, a high socio economic deprivation category and higher alcohol intake. Many patients had uncorrected CAD risk factors at pre-operative assessment. CONCLUSIONS: The SF-36 instrument was shown to be a useful and sensitive tool to assess differences and changes in the general health status of patients before and following CABG. High levels of social support were associated with improved health status post operatively. Lower pre-operative general health status, the presence of diabetes mellitus and cigarette smoking were associated with poorer post-operative general health status. PMID- 11053818 TI - Tricuspid valve replacement with the St. Jude Medical valve: 19 years of experience. AB - OBJECTIVE: The choice of the valve substitute in the tricuspid position remains controversial. A St. Jude Medical valve is a choice of valve substitute and its lower thrombogenicity and excellent hemodynamic performance have been reported even in the tricuspid position. However, little is known of the long-term durability of the St. Jude Medical valve in the tricuspid position. Our long-term experience of tricuspid valve replacement showed the higher thrombogenicity than we had expected, therefore, this study was done to reconsider our strategy for valve choice. METHODS: This study reviewed 23 patient who underwent 25 tricuspid valve replacements with the St. Jude Medical valves from 1980 to 1997. The mean age was 40 years. Eleven patients (48%) were men. There were four in-hospital deaths (17%). The remaining 19 patients were all alive and followed from 2.2 to 19.0 years (mean 11.8 years). RESULTS: The overall survival, including hospital mortality, was 83%, 10 and 15 years after surgery. Valve thrombosis occurred in six patients. Freedom from valve thrombosis was 78 and 70%, 10 and 15 years after surgery, respectively. The linearized rate of the valve thrombosis was 2.9%/patient-years. Six patients required reoperation. The mean interval to reoperation was 9.5 years. Freedom from reoperation was 83% and 75%, 10 and 15 years after surgery, respectively. The linearized rate of the reoperation was 2.8%/patient-years. No structural valve deterioration was found. Echocardiographic study showed that the function of the St. Jude Medical valve without valve-related complications was well maintained. CONCLUSIONS: The higher thrombogenicity of the St. Jude Medical valve in the tricuspid position altered our choice of valve substitutes from the St. Jude Medical valve to a bioprosthesis which is lack of need for anticoagulant therapy except for juvenile patients who are able to maintain potent anticoagulant therapy. PMID- 11053819 TI - Impact of duration of chest tube drainage on pain after cardiac surgery. AB - OBJECTIVE: This study was designed to analyze the duration of chest tube drainage on pain intensity and distribution after cardiac surgery. METHODS: Two groups of 80 cardiac surgery adult patients, operated on in two different hospitals, by the same group of cardiac surgeons, and with similar postoperative strategies, were compared. However, in one hospital (long drainage group), a conservative policy was adopted with the removal the chest tubes by postoperative day (POD) 2 or 3, while in the second hospital (short drainage group), all the drains were usually removed on POD 1. RESULTS: There was a trend toward less pain in the short drainage group, with a statistically significant difference on POD 2 (P=0.047). There were less patients without pain on POD 3 in the long drainage group (P=0. 01). The areas corresponding to the tract of the pleural tube, namely the epigastric area, the left basis of the thorax, and the left shoulder were more often involved in the long drainage group. There were three pneumonias in each group and no patient required repeated drainage. CONCLUSIONS: A policy of early chest drain ablation limits pain sensation and simplifies nursing care, without increasing the need for repeated pleural puncture. Therefore, a policy of short drainage after cardiac surgery should be recommended. PMID- 11053820 TI - Adult cardiac surgery outcomes: role of the pump type. AB - OBJECTIVE: This study was carried out to evaluate whether the type of pump used for cardiopulmonary bypass (CPB; roller vs. centrifugal) can affect mortality or the neurological outcomes of adult cardiac surgery patients. METHODS: Between 1994 and June 1999, 4000 consecutive patients underwent coronary and/or valve surgery at our hospital; of these, 2213 (55.3%) underwent surgery with centrifugal pump use, while 1787 (44.7%) were operated on with a roller pump. The effect of the type of the pump and of 36 preoperative and intraoperative risk factors for perioperative death, permanent neurological deficit and coma were assessed using univariate and multivariate analyses. RESULTS: The overall in hospital mortality rate was 2.2% (88/4000), permanent neurological deficit occurred in 2.0% (81/4000) of patients, and coma in 1.3% (52/4000). There was no difference in hospital mortality between patients operated with the use of centrifugal pumps and those operated with roller pumps (50/2213 (2.3%) vs. 38/1787 (2.1%); P=0.86). On the other hand, patients who underwent surgery with centrifugal pumps had lower permanent neurological deficit (34/2213, (1.5%) vs. 47/1787 (2.6%); P=0.020) and coma (20/2213 (0.9%) vs. 32/1787 (1.8%); P=0.020) rates than patients operated with roller pumps. Multivariate analysis showed CPB time, previous TIA and age as risk factors for permanent neurological deficit, while centrifugal pump use emerged as protective. Multivariate risk factors for coma were CPB time, previous vascular surgery and age, while centrifugal pump use was protective. CONCLUSIONS: Centrifugal pump use is associated with a reduced rate of major neurological complications in adult cardiac surgery, although this is not paralleled by a decrease in in-hospital mortality. PMID- 11053821 TI - Does aprotinin reduce lung reperfusion damage after cardiopulmonary bypass? AB - OBJECTIVE: The role of aprotinin in the prevention of lung reperfusion injury was investigated in the patients undergoing cardio-pulmonary bypass (CPB) for coronary artery bypass grafting (CABG) operations. METHODS: The study was planned randomly and prospectively. Two hundred milliliters of physiological saline solution was added to the prime solution of patients in group I (n=10) whereas, 200 ml aprotinin (Trasylol, Bayer AG) was given to patients in group II (n=10). In order to measure lung tissue malondialdehyde (MDA) levels, glutathion peroxidase (GSH-Px) activity levels and polymorphonuclear leukocytes (PMNs) numbers, lung tissue samples were taken before CPB and 5 min after removing the cross clamp. In addition, alveolo-arterial oxygen difference (AaDO(2)) for tissue oxygenation was calculated by obtaining arterial blood gas samples. RESULTS: MDA levels before CPB increased from 41.72+/-21.00 nmol/g tissue to 66.71+/-13.44 nmol/g tissue in group I and from 43.44+/-5.16 nmol MDA/g tissue to 53.22+/-10.95 nmol MDA/g tissue in group II after cross clamp removal (P=0.001 and P=0.021, respectively). The increase in group II was found to be significantly lower than group I (P=0.048). With the initiation of reperfusion, GSH-Px activity decreased in group I from 3.05+/-0.97 to 2.31+/-0.46 U/mg protein (P=0.015) whereas GSH-Px activity in group II decreased from 3.18+/-1.01 to 2.74+/-0.81 U/mg protein (P=0. 055). This decrease in the group II was less than group I (P=0.049). AaDO(2) significantly increased in the group I and II (P=0.012 and P=0.020, respectively), but elevation in the group I was significant than in the Group II (P=0.049). In histopathological examination, it was observed that neutrophil counts in the lung parenchyma rose significantly following removal of cross clamp in both groups (P=0. 001). The increase in group I was significantly larger than in group II (P=0.050). CONCLUSION: Results represented in our study indicate that addition of aprotinin (2 million units) into the prime solution during CPB can reduce lung reperfusion injury. PMID- 11053822 TI - Endothelial apoptosis is induced by serum of patients after cardiopulmonary bypass. AB - OBJECTIVE: Increased serum levels of a multitude of mediators like interleukins, tumor necrosis factor, elastase, adhesion molecules, and endotoxin have been described following cardiopulmonary bypass (CPB). The biological consequences of this complex response are unclear. METHODS: Serum samples of nine patients scheduled for elective coronary artery bypass grafting were obtained preoperatively and 1, 6, and 12 h after weaning from CPB. Additional serum samples were obtained perioperatively from four patients undergoing major lung resection and from four healthy volunteers. The apoptosis-inducing activity of serum samples on endothelial cells was examined using a tissue culture assay system. Endothelial cells were derived from human umbilical cords and incubated for 48 h with serum samples in various dilutions during their second passage. The culture plates were fixed with methanol/acetone and stained with the DNA dye diamidinophenylindole. Apoptotic and normal cells were identified and counted using phase contrast and fluorescence microscopy. RESULTS: The proportion of apoptotic endothelial cells was 5.6-fold higher in culture plates incubated with diluted (30%) serum samples obtained at 6 h after weaning from CPB when compared to plates incubated with preoperative samples (P=0.0077). A smaller effect occurred already at 1 h in some patients, whereas at 12 h after weaning from CPB no increased endothelial apoptosis was observed. No proapoptotic activity was found in preoperative as well as in control samples from patients undergoing lung resection or from healthy volunteers. CONCLUSIONS: Serum of patients after CPB exerts a strong apoptosis inducing activity on human endothelial cells. Apoptotic death of endothelial cells following CPB may be responsible for postoperative vascular and bypass dysfunction including phenomena like increased capillary permeability. PMID- 11053823 TI - Interleukin-1, interleukin-6 and myocardial enzyme response after coronary artery bypass grafting - a prospective randomized comparison of the conventional and three minimally invasive surgical techniques. AB - OBJECTIVE: In order to evaluate the traumatic effects of median sternotomy and cardiopulmonary bypass (CPB) in conventional and minimally invasive coronary artery bypass grafting, inflammatory response was studied in a prospective randomized trial in patients referred to single-vessel coronary artery bypass grafting. METHODS: Four surgical techniques were compared: group 1, median sternotomy with CPB in ten patients (eight male, two female; aged 59.6+/-11.0 years (mean+/-SD)); group 2, median sternotomy and off-pump in ten patients (seven male, three female; aged 65.1+/-10.0 years); group 3, minithoracotomy with CPB in ten patients (seven male, three female, aged 61.2+/-10.4 years); group 4, minithoracotomy and off-pump in ten patients (nine male, one female, aged 62.9+/ 9.8 years). All patients received a left internal mammary artery graft to the left anterior descending artery (LAD). Clinical data, perioperative values of cytokines and cardiac enzymes were monitored. RESULTS: There were no major complications. Troponin-T and creatine kinase isoenzyme MB (CK-MB) levels were significantly higher in CPB procedures (P<0.0056; multivariate general linear model). Interleukin-6 (IL-6) levels were significantly higher in minithoracotomy procedures. Interleukin-1 (IL-1) was significantly increased in all patients compared with the preoperative values. CONCLUSIONS: The use of CPB is combined with higher levels of troponin-T and CK-MB as signs of myocardial damage. Surgical access was identified as a trigger of inflammatory response, as minithoracotomy is related to higher levels of IL-6. IL-1 increased in all procedures and this occurred independently of the surgical access or the use of CPB, which points out a potential relationship between inflammatory response and anesthesia. Neither CPB nor surgical access influenced the clinical outcome in the treatment of coronary artery single-vessel bypass grafting. PMID- 11053824 TI - Phosphorylcholine coating of extracorporeal circuits provides natural protection against blood activation by the material surface. AB - OBJECTIVE: The aim of this study is to evaluate the use of a new coating, mimicking the outer cell membrane, in paediatric cardiac surgery. METHODS: Two groups of ten patients with a body weight below 8 kg, undergoing elective cardiac operations for different congenital anomalies, were prospectively enrolled in this study. In one group the whole extracorporeal circuit, including the cannulas, was coated with phosphorylcholine (PC). In the second group the same circuit was used without coating. Platelet activation (thromboxane B2 (TXB2), beta-thromboglobulin (betaTG)), activation of the coagulation system (F1+2), leukocyte activation (CD11b/CD18) and terminal complement activation (TCC) were analyzed pre-cardiopulmonary bypass (CPB), at 15, 60 min of CPB, at the end of CPB, 20 min post CPB and at postoperative day 1 and 6. RESULTS: No statistical differences were found for F1+2 and CD11b/CD18. After onset of CPB mean levels of TCC remained stable in the PC group whereas an increase was observed in the control group. During CPB betaTG values in both groups increased to a maximum at the end of CPB. Within groups the increase in betaTG levels during CPB was statistically significant (P<0.05) from baseline in the control group starting from 60 min of CPB whereas no statistical difference was observed in the PC group. After the start of CPB TXB2 mean levels increased to 405+/-249 pg/ml in the PC group vs. 535+/-224 pg/ml in the control group. After this initial increase there was a small decline in the PC group with further increase. This was in contrast to the control group were TXB2 levels further increased up to a mean of 718+/-333 pg/ml at the end of CPB (P=0.016). CONCLUSIONS: Phosphorylcholine coating had a favourable effect on blood platelets, which is most obvious after studying the changes during cardiopulmonary bypass. A steady increase of TXB2 and betaTG was observed in the control group, whereas plateau formation was observed in the phosphorylcholine group. Clinically, this effect may contribute to reduced blood loss and less thromboembolic complications. Complement activation is lower in the coated group. PMID- 11053825 TI - Ruptured papillary muscle. PMID- 11053826 TI - Bilateral open treatment of spontaneous pneumothorax: a new access. AB - A new technique for bilateral apical bullectomy and pleurectomy via axillary minithoracotomy and transmediastinal access to the contralateral side, was used in 13 patients with bilateral apical blebs and/or pneumothorax. The contralateral space is reached at the posterior superior mediastinum, passing between the first thoracic vertebral bodies (T1-T4) and the oesophagus. The contralateral lung apex is then pulled into the thoracotomy side and apical bullectomy carried out by linear stapler. The obvious advantages of avoiding a second thoracotomy while providing complete solution to the clinical problem are particularly important in young patients with spontaneous pneumothorax caused by bilateral apical blebs. PMID- 11053827 TI - A simple method to correct aortic tube graft kinking without cardiopulmonary by pass and aortic clamping. AB - A method for correcting the tube kinking after ascending aortic replacement for acute dissection is described. Its main advantage is the no need for cardiopulmonary by-pass (CPB) and aortic clamping to solve the problem. PMID- 11053828 TI - Internal thoracic artery as a collateral source to the ischemic lower extremity. AB - Based on the superior long-term results, internal thoracic artery is widely used for coronary artery bypass grafting. However, the vessel can play an important role as a collateral source to the chronically ischemic lower limbs. We reported two cases who underwent simultaneous revascularization to the myocardium and lower limbs because this particular condition was anticipated. Selective angiography of internal thoracic artery was useful to determine its role before harvesting in our cases. Careful preoperative examinations and choice of surgical approach are required for such patients to avoid serious vascular complications. PMID- 11053829 TI - Atherosclerotic disruption of the aortic arch during coronary artery bypass operation. AB - A 70-year-old-man presented with a symptomatic three vessel coronary artery disease and was scheduled for myocardial revascularization. During extracorporeal circulation an intrathoracal bleeding occurred and aortic rupture was suspected. An iatrogenic plaque rupture in the concavity of the aortic arch was found due to cannulation attempts. The aortic arch was grafted in the so-called elephant trunk technique. Thereafter bypass grafts were anastomosed to the stenosed coronary arteries. The patient was discharged from hospital after 2 weeks in good condition. PMID- 11053830 TI - The primitive neuroectodermal tumor of the heart. AB - A young man was admitted to hospital with dyspnea, malaise, chest pain and night sweating. Investigative studies revealed a cystic mass lesion originating from the heart. Surgical exploration of the tumor showed that it was unresectable and pathology of the biopsy material was primitive neuroectodermal tumor. Medical literature concerning this unusual type of tumor is reviewed. PMID- 11053831 TI - Intercostal arteriovenous hemangioma. AB - We report a case of a 46-year-old man who presented with a chest wall tumor in the right hemithorax. He underwent thoracotomy to remove the mass, which was found to be an arteriovenous hemangioma arising from the intercostal muscle. Arteriovenous hemangioma is a rare tumor and chest wall is an extremely rare site for this tumor. This tumor should be considered in the differential diagnosis of the chest wall tumors. Complete surgical excision offers the best treatment. PMID- 11053832 TI - Cloning, sequence and crystallographic structure of recombinant iron superoxide dismutase from Pseudomonas ovalis. AB - The gene encoding the iron-dependent superoxide dismutase from Pseudomonas ovalis was cloned from a genomic library and sequenced. The ORF differs from the previously published protein sequence, which was used for the original structure determination, at 16 positions. The differences include three additional inserted residues, one deleted residue and 12 point substitutions. The gene was subcloned and the recombinant protein overexpressed, purified and crystallized in a trigonal space group. The structure was determined by molecular replacement and was refined to 2.1 A resolution. PMID- 11053833 TI - Structure of XynB, a highly thermostable beta-1,4-xylanase from Dictyoglomus thermophilum Rt46B.1, at 1.8 A resolution. AB - Microorganisms employ a large array of enzymes to break down the cellulose and hemicelluloses of plant biomass. These enzymes, especially those with high thermal stability, have many uses in biotechnology. We have solved the crystal structure of a beta-1, 4-xylanase, XynB, from the extremely thermophilic bacterium Dictyoglomus thermophilum, isolate Rt46B.1. The protein crystallized from 1.6 M ammonium sulfate, 0.2 M HEPES pH 7.2 and 10% glycerol, with unit-cell parameters a = b = 91.3, c = 44.9 A and space group P4(3). The structure was solved at high resolution (1.8 A) by X-ray crystallography, using the method of isomorphous replacement with a single mercury derivative, and refined to a final R factor of 18.3% (R(free) = 22.1%). XynB has the single-domain fold typical of family 11 xylanases, comprising a jelly roll of two highly twisted beta-sheets that create a deep substrate-binding cleft. The two catalytic residues, Glu90 and Glu180, occupy this cleft. Compared with other family 11 xylanases, XynB has a greater proportion of polar surface and has a slightly extended C-terminus that, combined with the extension of beta-strand A5, gives additional hydrogen bonding and hydrophobic packing. These factors may account for the enhanced thermal stability of the enzyme. PMID- 11053834 TI - Structure of the C123S mutant of dienelactone hydrolase (DLH) bound with the PMS moiety of the protease inhibitor phenylmethylsulfonyl fluoride (PMSF). AB - The structure of DLH (C123S) with PMS bound was solved to 2.5 A resolution (R factor = 15.1%). PMSF in 2-propanol was delivered directly to crystals in drops and unexpectedly caused the crystals to dissolve. New crystals displaying a different morphology emerged within 2 h in situ, a phenomenon that appears to be described for the first time. The changed crystal form reflected altered crystal packing arrangements elicited by structural changes to the DLH (C123S) molecule on binding inhibitor. The new unit cell remained in the P2(1)2(1)2(1) space group but possessed different dimensions. The structure showed that PMS binding in DLH (C123S) caused conformational changes in the active site and in four regions of the polypeptide chain that contain reverse turns. In the active site, residues with aromatic side chains were repositioned in an edge-to-face cluster around the PMS phenyl ring. Their redistribution prevented restabilization of the triad His202 side chain, which was disordered in electron-density maps. Movements of other residues in the active site were shown to be related to the four displaced regions of the polypeptide chain. Their implied synergy suggests that DLH may be able to accommodate and catalyse a range of compounds unrelated to the natural substrate owing to an inherent coordinated flexibility in its overall structure. Implications for mechanism and further engineering studies are discussed. PMID- 11053835 TI - Structures of recombinant native and E202Q mutant human acetylcholinesterase complexed with the snake-venom toxin fasciculin-II. AB - Structures of recombinant wild-type human acetylcholinesterase and of its E202Q mutant as complexes with fasciculin-II, a 'three-finger' polypeptide toxin purified from the venom of the eastern green mamba (Dendroaspis angusticeps), are reported. The structure of the complex of the wild-type enzyme was solved to 2.8 A resolution by molecular replacement starting from the structure of the complex of Torpedo californica acetylcholinesterase with fasciculin-II and verified by starting from a similar complex with mouse acetylcholinesterase. The overall structure is surprisingly similar to that of the T. californica enzyme with fasciculin-II and, as expected, to that of the mouse acetylcholinesterase complex. The structure of the E202Q mutant complex was refined starting from the corresponding wild-type human acetylcholinesterase structure, using the 2.7 A resolution data set collected. Comparison of the two structures shows that removal of the charged group from the protein core and its substitution by a neutral isosteric moiety does not disrupt the functional architecture of the active centre. One of the elements of this architecture is thought to be a hydrogen-bond network including residues Glu202, Glu450, Tyr133 and two bridging molecules of water, which is conserved in other vertebrate acetylcholinesterases as well as in the human enzyme. The present findings are consistent with the notion that the main role of this network is the proper positioning of the Glu202 carboxylate relative to the catalytic triad, thus defining its functional role in the interaction of acetylcholinesterase with substrates and inhibitors. PMID- 11053836 TI - Structure of human alpha-thrombin complexed with RWJ-51438 at 1.7 A: unusual perturbation of the 60A-60I insertion loop. AB - The three-dimensional structure of the ternary complex consisting of human alpha thrombin, hirugen and the active-site inhibitor RWJ-51438 has been determined at 1.7 A resolution. The crystals of the complex belong to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 62.98, b = 117.52, c = 47.99 A. The refined R and R(free) values are 0.196 and 0.232, respectively. The ketone carbonyl group of the inhibitor is covalently linked to the hydroxyl O atom of Ser195, forming a tetrahedral intermediate hemiketal structure; the benzothiazole ring N atom of RWJ-51438 forms a hydrogen bond with His57. Surprisingly, the carboxylate substituent on the benzothiazole group forms salt bridges with Lys60F NZ and the NZ of the symmetry-related residues Lys236 and Lys240, which introduces steric effects that perturb the 60A-60I insertion loop, especially at residues Trp60D and Phe60H. PMID- 11053837 TI - The atomic resolution structure of bucandin, a novel toxin isolated from the Malayan krait, determined by direct methods. AB - Bucandin is a novel presynaptic neurotoxin isolated from Bungarus candidus (Malayan krait). It has the unique property of enhancing presynaptic acetylcholine release and represents a family of three-finger toxins with an additional disulfide in the first loop. There are no existing structures from this sub-category of three-finger toxins. The X-ray crystal structure of bucandin has been determined by the Shake-and-Bake direct-methods procedure. The resulting electron-density maps were of outstanding quality and allowed the automated tracing of 61 of the 63 amino-acid residues, including their side chains, and the placement of 48 solvent molecules. The 0.97 A resolution full-matrix least squares refinement converged to a crystallographic R factor of 12.4% and the final model contains 118 solvent molecules. This is the highest resolution structure of any member of the three-finger toxin family and thus it can serve as the best model for other members of the family. Furthermore, the structure of this novel toxin will help in understanding its unique ability to enhance acetylcholine release. The unique structure resulting from the fifth disulfide bond residing in the first loop improves the understanding of other toxins with a similar arrangement of disulfide bonds. PMID- 11053838 TI - Crystallographic studies of the interaction between the ferredoxin-NADP+ reductase and ferredoxin from the cyanobacterium Anabaena: looking for the elusive ferredoxin molecule. AB - Ferredoxin-NADP(+) reductase (FNR) and its physiological electron donor ferredoxin (Fd) from the cyanobacterium Anabaena PCC7119 have been co crystallized. The unit-cell parameters are a = b = 63.72, c = 158.02 A and the space group is P2(1)2(1)2(1). The crystal structure has been solved with 2.4 A resolution synchrotron data by molecular replacement, anomalous dispersion and R(min) search methods. For the computations, the crystal was treated as a merohedral twin. The asymmetric unit contains two FNR molecules and one ferredoxin molecule. The packing of the FNR molecules displays a nearly tetragonal symmetry (space group P4(3)2(1)2), whereas the ferredoxin arrangement is orthorhombic. This study provides the first crystallographic model of a dissociable complex between FNR and Fd. PMID- 11053839 TI - Single-wavelength anomalous diffraction phasing revisited. AB - Multiwavelength anomalous diffraction (MAD) phasing has become a routinely used tool for determining new macromolecular structures. The MAD method has stringent data-collection requirements, typically necessitating radiation-resistant crystals and access to a tunable synchrotron beamline. In cases where synchrotron time, monochromator tunability or radiation damage is a concern or where high throughput structure determination is desired, phasing methods capable of producing interpretable electron-density maps from less data become attractive alternatives to MAD. The increasing availability of tunable synchrotron data collection facilities prompted the authors to revisit single-wavelength anomalous diffraction (SAD) phasing used in conjunction with a phase-ambiguity resolving method such as solvent flattening. The anomalous diffraction from seven different selenomethionine-labelled protein crystals has been analysed and it is shown that in conjunction with solvent flattening, diffraction data from the peak anomalous wavelength alone can produce interpretable electron-density maps of comparable quality to those resulting from full MAD phasing. Single-wavelength anomalous diffraction (SAD) phasing can therefore be a time-efficient alternative to MAD. The data also show that radiation damage can have a significant effect on the quality of SAD/MAD diffraction data. These results may be useful in the design of optimal strategies for collection of the diffraction data. PMID- 11053840 TI - Evaluation of phase accuracy via topological and geometrical analysis of electron density maps. AB - An empirical function is developed to measure the protein-like character of electron-density maps. The function is based upon a systematic analysis of numerous local and global map properties or descriptors. Local descriptors measure the occurrence throughout the unit cell of unique patterns on various defined templates, while global descriptors enumerate topological characteristics that define the connectivity and complexity of electron-density isosurfaces. We examine how these quantitative descriptors vary as error is introduced into the phase sets used to generate maps. Informative descriptors are combined in an optimal fashion to arrive at a predictive function. When the topological and geometrical analysis is applied to protein maps generated from phase sets with varying amounts of error, the function is able to estimate changes in average phase error with an accuracy of better than 10 degrees. Additionally, when used to monitor maps generated with experimental phases from different heavy-atom models, the analysis clearly distinguishes between the correct heavy-atom substructure solution and incorrect heavy-atom solutions. The function is also evaluated as a tool to monitor changes in map quality and phase error before and after density-modification procedures. PMID- 11053841 TI - Evaporative microdialysis: an effective improvement in an established method of protein crystallization. AB - Evaporative dialysis is a simple variant of conventional microdialysis in which the reservoir solution is allowed to evaporate slowly. The slow increase in precipitant concentration allows crystals to grow without increasing nucleation. The method is useful for proteins that have a very narrow metastable zone (the range of solution conditions under which crystals grow but nuclei do not form at an appreciable rate). The method is demonstrated with the coat protein of potato virus X. PMID- 11053842 TI - Crystallization and preliminary crystallographic study of an extremophile cytochrome c4 from Thiobacillus ferrooxidans. AB - Soluble periplasmic dihaemic cytochrome c(4), of 21 293 Da molecular mass, has been characterized from Thiobacillus ferrooxidans, an acidophilic bacteria. The native cytochrome has been purified from the bacteria using ion-exchange chromatography and crystallized using solution 27 of the Hampton Research Crystal Screen II. The crystals belong to the hexagonal space group P6(2)22 or P6(4)22, with unit-cell parameters a = 101.59, b = 101.59, c = 151.59 A. Frozen crystals diffract to 2.17 A resolution. The MAD method is currently being used (four Fe atoms per asymmetric unit) to solve the protein structure. PMID- 11053843 TI - Crystallization and preliminary X-ray diffraction analysis of a eumenine mastoparan toxin: a new class of mast-cell degranulating peptide in the wasp venom. AB - Mastoparans are tetradecapeptides found to be the major component of vespid venoms. A mastoparan toxin isolated from the venom of Anterhynchium flavomarginatum micado has been crystallized and X-ray diffraction data collected to 2.7 A resolution using a synchrotron-radiation source. Crystals were determined to belong to the space group P6(2)22 (P6(4)22). This is the first mastoparan to be crystallized and will provide further insights into the conformational significance of mastoparan toxins with respect to their potency and activity in G-protein regulation. PMID- 11053844 TI - Preliminary crystallographic studies of an extremely thermostable KDG aldolase from Sulfolobus solfataricus. AB - Crystals have been grown of 2-keto-3-deoxygluconate aldolase (KDG aldolase) from the hyperthermophilic archaeon Sulfolobus solfataricus that diffract to 2.2 A resolution. The enzyme catalyses the reversible aldol cleavage of 2-keto-3 dexoygluconate to pyruvate and glyceraldehyde, the third step of a modified non phosphorylated Entner-Doudoroff pathway of glucose oxidation. S. solfataricus grows optimally at 353 K and the enzyme itself has a half-life of 2.5 h at 373 K. Knowledge of the crystal structure of KDG aldolase will further understanding of the basis of protein hyperthermostability and create a target for site-directed mutagenesis of active-site residues, with the aim of altering substrate specificity. Three crystal forms have been obtained: orthorhombic crystals of space group P2(1)2(1)2(1), which diffract to beyond 2.15 A, monoclinic crystals of space group C2, which diffract to 2.2 A, and cubic crystals of space group P4(2)32, which diffract to 3.4 A. PMID- 11053845 TI - Crystallization and preliminary X-ray diffraction analysis of a recombinant cysteine-free mutant of crmA. AB - CrmA is an unusual serpin that has a reactive-center loop one residue shorter than other members of the superfamily. Most interestingly, crmA has inhibitory activity against both cysteine and serine proteinases involved in the regulation of cell apoptosis. The three-dimensional structure of crmA will give insight into the mechanism that this serpin employs to inhibit both cysteine and the serine proteinases, as well as help to explain the significance of the shorter reactive center loop. The monodisperse cysteine-free mutant of crmA was crystallized in the presence of phosphate salts. Crystals diffract to 2.90 A and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 42.67, b = 93.15, c = 101.63 A. PMID- 11053846 TI - Crystallization and preliminary X-ray analysis of glucose dehydrogenase from Bacillus megaterium IWG3. AB - Glucose dehydrogenase from Bacillus megaterium IWG3 has been crystallized in the presence of NAD(+) using the hanging-drop vapour-diffusion method with PEG 2000 as the precipitant. Crystals belong to space group C2 and have unit-cell parameters a = 120.8 (1), b = 66.7 (1), c = 119.6 (1) A, beta = 93.25 (3) degrees with standard deviations in parentheses. Assumption of four subunits in the asymmetric unit gave the most probable Matthews coefficient V(M) of 2.1 A(3) Da( 1) (solvent content 41.7% by volume). X-ray diffraction data were collected to 1.7 A on a synchrotron-radiation source. PMID- 11053847 TI - Crystallization and preliminary X-ray crystallographic studies of response regulator for cyanobacterial phytochrome, Rcp1. AB - The key response-regulator gene of light regulation, rcp1, from Synechocystis sp. has been overexpressed, purified and subsequently crystallized using ammonium sulfate as a precipitant in forms suitable for X-ray crystallographic studies. A native data set was collected to a resolution of 2.5 A at cryogenic temperature. The crystals belong to the hexagonal space group P6(3), with unit-cell parameters a = b = 89.04 (5), c = 60.29 (3) A. The Matthews parameter suggests that Rcp1 crystallizes with two molecules per asymmetric unit. PMID- 11053848 TI - Expression, crystallization and preliminary X-ray diffraction studies of recombinant Bacillus anthracis lethal factor. AB - The lethal factor (LF) produced by Bacillus anthracis is a Zn(2+)-dependent endopeptidase which specifically cleaves the N-terminal tail of several MAP kinase kinases (MAPKKs). The recombinant expression, purification and crystallization of LF and of an inactive mutant consisting of a single amino-acid substitution in the conserved catalytic site are reported here. Both proteins crystallize in the cubic space group I432. PMID- 11053849 TI - Crystallization and preliminary X-ray diffraction analysis of a functional form of pneumolysin, a virulence factor from Streptococcus pneumoniae. AB - Pneumolysin is a virulence factor from Streptococcus pneumoniae, a Gram-positive bacterial pathogen which causes human infections with a severe impact on mortality and morbidity worldwide. The enzyme belongs to a group of cholesterol dependent cytolysins and interacts with its cholesterol receptor on target cells, leading to pneumolysin insertion into target-cell membranes and subsequently to pore formation and cell lysis. Pneumolysin has been overexpressed, purified and crystallized for X-ray diffraction studies. Crystals have been obtained in the presence of cholesterol in an effort to produce a three-dimensional structure of pneumolysin in its fully functional form with the enzyme bound to its activator. This is the first report of the crystallization of a cholesterol-dependent cytolysin in the presence of bound cholesterol. The vapor-diffusion method using ammonium sulfate as a precipitation agent was used to grow crystals in the presence of n-octyl-beta-D-glucopyranoside and phosphatidylcholine. Crystals of this 53 kDa molecule complexed with cholesterol diffracted X-rays to 3.3 A. The crystal unit cell has parameters a = b = 191.45, c = 66.16 A, alpha = beta = 90.0, gamma = 120 degrees and belongs to the trigonal space group P3. The determination of the three-dimensional structure of this pneumococcal cytolysin is in progress. PMID- 11053850 TI - Crystallization and preliminary X-ray diffraction studies on the DNA-binding domain of the multidrug transporter activation protein (MtaN) from Bacillus subtilis. AB - The N-terminal DNA-binding domain of the multidrug transporter activation protein (MtaN) was crystallized by the hanging-drop vapour-diffusion method using lithium chloride as a precipitant. The crystals are orthorhombic and belong to the space group I2(1)2(1)2(1), with unit-cell parameters a = 49.4, b = 67.8, c = 115. 0 A. Diffraction data have been collected at 100 K to 2.75 A resolution at a synchrotron-radiation source. PMID- 11053851 TI - Crystallization and preliminary X-ray analysis of full-length annexin I comprising the core and N-terminal domain. AB - Annexin I, a member of the annexin family of Ca(2+)- and phospholipid-binding proteins, has been crystallized with the complete N-terminus. Annexins are structurally divided into a conserved protein core and an N-terminal domain that is variable in sequence and length. Three-dimensional structures of annexins comprising the protein core and a short N-terminal domain (annexins III, IV, V, VI, XII) or a truncated form almost completely lacking the N-terminal domain (annexins I and II) have been published so far. Here, the crystallization of annexin I comprising not only the core but also the complete N-terminal domain is reported. The crystals belong to the space group P2(1)2(1)2(1), with unit-cell parameters a = 63.6, b = 96.3, c = 127.4 A, and diffract to better than 2 A. Assuming a molecular weight of 38.7 kDa for annexin I and an average value of 2.5 A(3) Da(-1) for V(M), two molecules per asymmetric unit are present. PMID- 11053852 TI - Crystallization and preliminary X-ray analysis of a bacterial lysozyme produced by Streptomyces globisporus. AB - The extracellular bacteriolytic enzyme produced by Streptomyces globisporus shows a beta-1,4-N,6-O-diacetylmuramidase activity as well as a beta-1,4-N acetylmuramidase activity. Crystals of this enzyme have been obtained by the hanging-drop vapour-diffusion method using polyethylene glycol as a precipitant. They belong to the tetragonal space group P4(1)2(1)2, with unit-cell parameters a = 63.11 (4), c = 121.1 (1) A, diffract to at least 2.0 A resolution and are suitable for high-resolution structure analysis. The crystal structure was solved by molecular replacement using lysozyme produced by S. erythraeus as a search model. The structure refinement is now in progress. PMID- 11053853 TI - Expression, purification and crystallization of human tau-protein kinase I/glycogen synthase kinase-3beta. AB - Human tau-protein kinase I (TPK-I; also known as glycogen synthase kinase-3beta, GSK-3beta) is a serine/threonine protein kinase. Full-length TPK-I/GSK-3beta was expressed in Escherichia coli as a fusion protein with a 6xHis tag at the C terminus and was crystallized using the hanging-drop vapour-diffusion method. Prismatic crystals of dimensions 0.4 x 0.2 x 0.1 mm were obtained using 12 15%(w/v) polyethylene glycol 6000 as a precipitant at 278 K. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 82.9, b = 86.1, c = 178.1 A measured at 100 K, diffract to 2.3 A resolution and seem to contain two enzyme molecules per asymmetric unit. PMID- 11053854 TI - Crystallization and preliminary crystallographic studies of trichomaglin, a novel ribosome-inactivating protein. AB - Trichomaglin, a novel ribosome-inactivating protein, has been crystallized in two crystal forms using the hanging-drop vapour-diffusion method. The form A and form B crystals belong to the orthorhombic space group P2(1)2(1)2(1) and the hexagonal space group P6(1) (or P6(5)), respectively. X-ray data have been collected to 3.3 and 2.2 A resolution for the form A and B crystals, respectively. PMID- 11053855 TI - Crystallization and preliminary X-ray analysis of CTLA-4 (CD152) membrane external domain. AB - CTLA-4 (CD152) is involved in T-lymphocyte co-stimulatory pathways modulating both humoral and cellular immune response. The membrane-external domain has been prepared and crystallized. The unit-cell parameters are a = b = 43, c = 143 A with the symmetry of space group P3(1)21 or its enantiomer and the crystals diffract to 2. 7 A resolution at synchrotron beamlines. PMID- 11053856 TI - Crystallization and preliminary X-ray crystallographic studies of the thermoactive pullulanase type I, hydrolyzing alpha-1,6 glycosidic linkages, from Fervidobacterium pennivorans Ven5. AB - Crystals of the thermoactive recombinant F. pennivorans type I pullulanase, purified from the supernatant of a Bacillus subtilis culture, have been obtained by the vapour-diffusion method in the presence of the inhibitor beta-cyclodextrin (2 mM) by mixing protein (15 mg ml(-1)) with an equal volume of crystallization solution containing 0.1 M bis-tris propane pH 6.5, 50 mM MgCl(2) and 15% polyethylene glycol 3350. Crystals diffracted to 3.0 A using conventional Cu Kalpha radiation and belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 76.8, b = 96.2, c = 98. 5 A. The asymmetric unit contains one monomer. A preliminary 26% complete data set has been collected at 2.2 A resolution using synchrotron radiation. PMID- 11053857 TI - Crystallization and preliminary X-ray crystallographic studies on the bacteriophage phi6 RNA-dependent RNA polymerase. AB - The RNA-dependent RNA polymerase (P2) from bacteriophage Phi6 has been cloned and the protein overexpressed in Escherichia coli to produce an active enzyme. A fully substituted selenomethionyl version of the protein has also been produced. Crystals of both proteins have been grown; most belong to the monoclinic space group P2(1), with unit-cell parameters a = 105.9, b = 94.0, c = 140.9 A, beta = 101.4 degrees, but some are trigonal (space group P3(1) or P3(2)), with unit-cell parameters a = b = 110.1, c = 159.4 A, gamma = 120 degrees. Both crystal forms occur in the same crystallization drop and are morphologically indistinguishable. Native data sets have been collected from both types of crystals to better than 3 A resolution. PMID- 11053858 TI - Crystallization and preliminary X-ray analysis of the auxin receptor ABP1. AB - Auxin-binding protein (ABP1) is an important receptor for the plant hormone auxin that is involved with many growth and developmental responses in plants. The maize ABP1 gene has been expressed in insect cells, purified and crystallized. Type II crystals are monoclinic, with two glycosylated homodimers in the asymmetric unit, and diffract to 1.9 A using synchrotron radiation. PMID- 11053859 TI - Purification, crystallization and preliminary X-ray crystallographic analysis of human CCG1-interacting factor B. AB - A novel human factor CIB (CCG1-interacting factor B) has been isolated using the yeast two-hybrid system. The 22 kDa CIB protein has been expressed in Escherichia coli, purified to homogeneity and crystallized in a form suitable for crystallographic studies. The protein was crystallized in the orthogonal space group P2(1)2(1)2(1), with unit-cell parameters a = 43.60 (2), b = 44.45 (1), c = 110.70 (5) A and one molecule in the asymmetric unit. The crystal diffracted beyond 2.2 A resolution using synchrotron radiation. PMID- 11053860 TI - Crystallization and preliminary X-ray diffraction studies of a new crystal form of human secretory type IIA phospholipase A2. AB - Human synovial type IIA phospholipase A(2) (sPLA(2)-IIA) has been implicated in the pathogenesis of a number of inflammatory diseases and is a target for the development of therapeutically useful inhibitors. Biochemical evidence suggests a novel mechanism of inhibition for a series of peptide inhibitors originally derived from the primary sequence of the protein. On co-incubation with one of these inhibitors, single crystals of a hitherto unreported crystallographic form of sPLA2-IIA suitable for diffraction analysis were obtained. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 140.8, b = 38.9, c = 109.1 A, beta = 125.1 degrees, and diffraction at 2.4 A resolution has been observed. PMID- 11053861 TI - Nucleoside diphosphate kinase from the hyperthermophilic archaeon Methanococcus jannaschii: overexpression, crystallization and preliminary X-ray crystallographic analysis. AB - Nucleoside diphosphate (NDP) kinase is a key enzyme in maintaining cellular pools of all nucleoside triphosphates. NDP kinase from the hyperthermophilic archaebacterium Methanococcus jannaschii has been overexpressed in Escherichia coli and crystallized at 297 K using polyethylene glycol 4000 as precipitant. The crystal is hexagonal, belonging to the space group P6(3), with unit-cell parameters a = b = 72.89, c = 100.87 A. The asymmetric unit contains two subunits of NDP kinase, with a corresponding crystal volume per protein mass (V(M)) of 2.38 A(3) Da(-1) and a solvent content of 48.3%. Native X-ray diffraction data to 2.30 A resolution have been collected using synchrotron X-rays. PMID- 11053862 TI - Preliminary X-ray diffraction studies of rabbit muscle triose phosphate isomerase (TIM). AB - Triose phosphate isomerase (TIM) is responsible for the interconversion between GAP and DHAP in the glycolytic pathway. Two crystal forms belonging to space group P2(1)2(1)2(1) were obtained by the hanging-drop method and were designated A and B. Synchrotron X-ray diffraction data were collected for both forms. Form A had unit-cell parameters a = 65.14, b = 72.45, c = 93.24 A and diffracted to 2.25 A at 85 K, whereas form B had unit-cell parameters a = 73.02, b = 79.80, c = 172.85 A and diffracted to 2.85 A at room temperature. Molecular replacement was employed to solve the structures, using human TIM as a search model. Further refinement of both structures is under way and is expected to shed light on the recently reported conformational studies for rabbit TIM. PMID- 11053863 TI - Cloning, expression, purification and crystallization of dihydroxybutanone phosphate synthase from Magnaporthe grisea. AB - Dihydroxybutanone phosphate synthase (DS) catalyzes a commitment step in riboflavin biosynthesis where ribulose 5-phosphate is converted to dihydroxybutanone phosphate and formate. DS was cloned from the pathogenic fungus Magnaporthe grisea (using functional complementation of an Escherichia coli DS knockout mutant) and expressed in E. coli. The purified protein crystallized in space group P2(1)2(1)2. Diffraction data extending to 1.5, 1.0 and 1.8 A resolution were collected from crystals that were divalent cation free, soaked in Zn(2+) or soaked in Mg(2+), respectively. PMID- 11053864 TI - Crystallization and preliminary X-ray studies of oligandrin, a sterol-carrier elicitor from Pythium oligandrum. AB - Oligandrin is a 10 kDa acidic protein produced by the fungus micromycete Pythium oligandrum and is a member of the alpha-elicitin group, with sterol- and lipid carrier properties. Oligandrin has been crystallized at 290 K using PEG 4000 as a precipitant. A cholesterol complex was obtained under the same conditions. The space group of the crystals at low temperature (100 K) is C222, with unit-cell parameters a = 94.0, b = 171.1, c = 55.3 A. Four molecules are present in the asymmetric unit. Data from the free and cholesterol-complexed forms were recorded at synchrotron sources to resolutions of 2.4 (uncomplexed) and 1.9 A (complexed), respectively. PMID- 11053865 TI - The purification, crystallization and preliminary structural characterization of glucose-1-phosphate thymidylyltransferase (RmlA), the first enzyme of the dTDP-L rhamnose synthesis pathway from Pseudomonas aeruginosa. AB - Glucose-1-phosphate thymidylyltransferase (RmlA; E.C. 2.7.7.24) is the first of four enzymes involved in the biosynthesis of dTDP-L-rhamnose, the precursor of L rhamnose, a key component of the cell wall of many pathogenic bacteria. RmlA catalyses the condensation of thymidine triphosphate (dTTP) and alpha-D-glucose-1 phosphate (G1P), yielding dTDP-D-glucose. RmlA from Pseudomonas aeruginosa has been overexpressed and purified. Crystals of the enzyme have been grown using the sitting-drop vapour-diffusion technique with PEG 6000 and lithium sulfate as precipitant. Several diffraction data sets of single frozen crystals were collected to a resolution of 1.66 A. Crystals belonged to space group P1, with unit-cell parameters a = 71.5, b = 73.1, c = 134.7 A, alpha = 89.9, beta = 80.9, gamma = 81.1 degrees. The asymmetric unit contains eight monomers in the form of two RmlA tetramers with a solvent content of 51%. Selenomethionine-labelled protein has been obtained and crystallized. PMID- 11053866 TI - Crystallization and preliminary crystallographic studies of a new L-asparaginase encoded by the Escherichia coli genome. AB - A new Escherichia coli L-asparaginase belonging to the class of Ntn amidohydrolases has been crystallized using the vapour-diffusion method and PEG 4000 as the precipitant. The crystals belong to the orthorhombic space group P2(1)2(1)2(1) (unit-cell parameters a = 50. 3, b = 77.6, c = 148.2 A) and diffract to 1.65 A resolution. The structure has been solved by molecular replacement using aspartylglucosaminidase from Flavobacterium meningosepticum as the search model. The asymmetric unit contains four protein chains composed into a dimer of alphabeta heterodimers, where the subunits alpha and beta are the product of autoproteolytic cleavage of the immature protein. PMID- 11053867 TI - Purification, crystallization and preliminary diffraction study of beta galactosidase from Penicillium sp. AB - Crystals of an extracellular beta-galactosidase from Penicillium sp. (MW = 120 +/ 5 kDa) have been obtained from a sodium phosphate buffer using PEG as precipitant. The crystals belong to the tetragonal space group P4(1)or P4(3), with unit-cell parameters a = b = 110.82, c = 161.28 A, and diffract to 1.85 A resolution at a synchrotron source. PMID- 11053868 TI - Apotheosis, not apocalypse: methods in protein crystallography. PMID- 11053869 TI - Crystallization and preliminary X-ray diffraction studies of monomeric isocitrate dehydrogenase from corynrbacterium glutamicum. erratum AB - In the paper by Audette et al. [Acta Cryst. (1999), D55, 1584-1585] the postal code of one of the authors was printed incorrectly. The correct version is given above. Also the reaction catalyzed by isocitrate dehydrogenase was given incorrectly in the paper; the correct reaction is given below. Isocitrate + NADP(+) <--> Oxalosuccinate + NADPH. Oxalosuccinate <--> alpha-Ketoglutarate + CO(2). PMID- 11053870 TI - Crystallization and preliminary X-ray analysis of the conserved domain IV of escherichia coli 4.5S RNA. erratum AB - In the paper by Jovine et al. [Acta Cryst. (2000), D56, 1033-1037] the name of the second author was given incorrectly. The correct name should be Tobias Hainzl as given above. PMID- 11053871 TI - The integration of family planning and genitourinary medicine services. PMID- 11053872 TI - Peer review at the british journal of family planning PMID- 11053873 TI - Knowledge of genital Chlamydia trachomatis infection in family planning clinic attenders. AB - The purpose of this study was to determine the level of awareness of genital Chlamydia infection and level of knowledge related to this infection in family planning (FP) clinic attenders. Clients attending FP clinics during a 3 month study period were invited to complete an anonymous self-administered questionnaire. Five hundred and sixteen questionnaires from female attenders were analysed. Results showed that 54% of respondents had heard of Chlamydia. Subjective knowledge assessment for Chlamydia was low compared to that for other infections. Mean knowledge scores relating to genital chlamydial infection were low. There was no significant age-related trend in knowledge scores. The implications of these findings are discussed in relation to increased Chlamydia screening activity in FP clinics. PMID- 11053874 TI - Chlamydia trachomatis screening in young people in Merseyside. AB - OBJECTIVES: To evaluate the acceptability to young people of proactive Chlamydia trachomatis (CT) information and urine test. To discover the extent of CT infection and the practical implications for completing treatment and partner notification. DESIGN: Prospective screening with sexual health questionnaire. SETTING: Three family planning clinics for young people in Liverpool and South Sefton. PARTICIPANTS: Nine hundred and five women and 53 men had urine tests and answered the questionnaire. All aged 20 years or under attending the clinics were given information about CT and safer sex. MAIN OUTCOME MEASURES: The acceptability of proactive information and screening for CT using a urine test. Prevalence of CT infection. The time and effort incurred informing and managing those testing positive. RESULTS: The information and urine test were readily accepted. Prevalence of CT was 8.5% in women and 5.7% in men. More than three quarters of those testing positive were treated, but it took much time and effort, as follow-up attendance was poor. CONCLUSIONS: The prevalence of CT was high in this population. Young people participated in screening readily. They are interested in this health issue, but it was difficult to hold their attention long enough to complete the process of treatment and contact tracing. Completing this successfully either needs a huge input of resources or a new approach. These results have led to the piloting of an outreach health adviser administering treatment and carrying out partner notification at the screening site. Some of the questions raised by the CMO have been addressed. PMID- 11053875 TI - Feasibility of patient-collected vulval swabs for the diagnosis of Chlamydia trachomatis in a family planning clinic: a pilot study. AB - This pilot study set out to determine the feasibility of using patient-collected vulval swabs, instead of urine, for the diagnosis of female Chlamydia trachomatis infection. Main outcome measures included prevalence of infection and sensitivity, specificity, and acceptability of both test methods. An assessment was also made of those who declined to be tested. Consecutive women under 25 years of age attending a single urban family planning clinic were invited to participate. Sixty-eight percent (103/152) agreed to undergo testing. Overall prevalence was 11.7%. The sensitivity/specificity for the ligase chain reaction (LCR) assayed patient-collected vulval swabs and urine was 100%/100% and 92%/100%, respectively. The acceptability of self-collection was high with 93% characterising the test as 'not bad', 79% recommending it to a friend, and 79% choosing the test next time. Significantly more women, however, would choose urine for testing on a subsequent occasion (p < 0.001). Less than 1/5 of the patients who declined did not take part because of concerns regarding the vulval swab. Patient-collected vulval swabs assayed by LCR represent a non-invasive, sensitive, and acceptable way to detect genital C. trachomatis infection in women attending a family planning clinic. Compared with urine testing, benefits in terms of transport and processing should encourage more widespread use of this approach. PMID- 11053876 TI - An interface of chlamydia testing by community family planning clinics and referral to hospital genitourinary medicine clinics. AB - OBJECTIVES: To assess compliance with the protocol for the management of women with Chlamydia trachomatis diagnosed in community family planning (FP) clinics; to assess the rate of attendance at genitourinary medicine (GUM) clinics by these women; to assess the rate of adequate treatment and to assess the level of communication between GUM clinics and FP clinics. METHOD: Retrospective review of FP clinic records and case notes to identify all women with positive or equivocal Chlamydia results during a 6 month period, and a retrospective review of records from five local GUM clinics. RESULTS: One hundred and twelve women were identified from FP clinic records with positive or equivocal Chlamydia results. Eighty-nine (79.5%) were referred to a GUM clinic. Twelve out of 14 women not referred had equivocal results. The median delay from the test being taken to the results being seen by a doctor was 9 days, and to the woman being referred was 10 days. Fifty-eight (51.7%, n = l12) women definitely attended a local GUM clinic. The FP clinics provided a letter of referral in 76 (85.4%, n = 89) women and the GUM clinics provided a letter of reply in 21 (48.8%, n = 43) women who attended with a referral letter. Three months after testing, only 54 (48.2%) of the 112 women with positive or equivocal Chlamydia tests were known by the referring FP clinic to have been treated. CONCLUSIONS: The majority of women with positive or equivocal Chlamydia results were referred to a GUM clinic according to the protocol. Attendance at GUM clinics was disappointing, as only 51.7% of the 112 women with positive or equivocal results had documented evidence of having attended. This raises the question not whether community clinics should be testing, but whether they should be initiating treatment and partner notification. Collaborative work between GUM clinics and community clinics around partner notification is needed, as well as funding for training and additional pharmacy costs. Further collaborative work between GUM and FP and reproductive healthcare (RHC) to evaluate the role of community clinics in the diagnosis and management of chlamydial infection and other sexually transmitted infections (STIs) is needed. PMID- 11053877 TI - An exploratory study of information-giving used to promote chlamydial test seeking by students at a college family planning clinic. AB - Partnership working, involving workers in various aspects of sexual health and a large UK further education college, took place to give information about genital tract chlamydial infection in order to promote chlamydial urine testing (LC(x) Chlamydia trachomatis Assay Abbott Diagnosis Division) for a limited period at the college's family planning clinic. Female students were more likely to report awareness about the availability of testing and to access the testing service. Uptake of testing was largely contemporaneous with information-giving work and sharply declined after information-giving had ceased. A small population of test seekers (including partners of index cases) was generated, which harvested a rate of genital tract chlamydial infection similar to that found in family planning and genitourinary medicine clinics. PMID- 11053878 TI - Concepts of compliance: understandings and approaches. AB - This paper will take a critical look at the concepts of compliance and non compliance, both generally and in the specific context of contraceptive use. PMID- 11053879 TI - News online PMID- 11053880 TI - Integrating family planning with genitourinary medicine: developing an holistic sexual health clinic in Eastbourne. AB - The idea of providing family planning and genitourinary medicine under one roof has attracted much interest. The development of an integrated sexual health clinic in Eastbourne is described, from initial discussions between disparate parties to the emergence of a one-stop-shop, with a look to the future. PMID- 11053881 TI - Magnetic resonance imaging appearances of the Mirena and GyneFix intra-uterine contraceptive devices: a report of two cases. AB - We report here two cases which illustrate the magnetic resonance imaging (MRI) appearance of the Mirena levonorgestrel releasing intra-uterine system and the GyneFix copper intra-uterine contraceptive implant. The MRI appearance of these devices has not to our knowledge been reported to date, and as increasing numbers of women choose to use these devices for treatment of gynaecological conditions and contraception, it becomes increasingly important to recognise their appearance on pelvic imaging. PMID- 11053882 TI - First-line management of menorrhagia: findings from a survey of general practitioners in Forth Valley. AB - This study was undertaken to determine local GPs' current management of menorrhagia, prior to the release of the Royal College of Obs tetricians and Gynaecologists' (RCOG) guideline on the initial management of menorrhagia. A postal questionnaire was sent to 204 GPs in the Forth Valley area. An 84% (n = 173) response rate was recorded. Responses were compared with RCOG guideline recommendations after its release. Fifty-two percent of respondents measured a full blood count and 56% performed a pelvic examination. Two thirds of GPs chose the recommended treatments (mefenamic acid or tranexamic acid) as their first or second choice of treatment in women not requiring contraception. One third chose cyclical progestogens. Only 10% of GPs selected an ineffective treatment (cyclical progestogens) as their first or second choice of treatment in women requiring contraception. This survey demonstrates that the majority of GPs are already prescribing the treatments recommended in the RCOG guidelines. There is a need for education amongst a minority to improve their practice. Half of the GPs reported compliance with the recommended investigations of full blood count and pelvic examination. The point at which these investigations are performed in the initial management of menorrhagia may need further discussion between primary and secondary care clinicians to clarify their purpose. PMID- 11053884 TI - 'Last trance' baby experts register a first; FDA approves new contraception cap; licensed to chill; quitting smoking quantified; medical staff join the dot.com revolution PMID- 11053883 TI - Hysteroscopic management of intra-uterine devices with lost strings. AB - We report a series of 38 patients with intra-uterine devices with lost strings where hysteroscopic aid was required after routine retrieval procedures failed. Thirty-five intra-uterine devices could be removed easily with hysteroscope. In one patient a fragmented Lippes Loop was removed piecemeal hysteroscopically. Laparotomy was required in only one patient, for an extra-uterine Copper T. Hysteroscopy is thus a simple, safe and effective method for removing misplaced intra-uterine devices. PMID- 11053885 TI - My MFFP dissertation. AB - This paper describes how Amanda Smith set about her MFFP dissertation, from start to finish, and asks the question 'Was it all worth it?' PMID- 11053886 TI - Report from the faculty of family planning and reproductive health care AGM, May 2000. PMID- 11053887 TI - Combined oral contraception and cancer. PMID- 11053888 TI - Question sheet/ answer sheet PMID- 11053889 TI - Notes for contributors PMID- 11053890 TI - From the journals PMID- 11053892 TI - Meetings and courses PMID- 11053891 TI - Anti-D guidelines. PMID- 11053893 TI - Fees for DFFP practical training sessions. PMID- 11053894 TI - Amelioration of alloxan induced diabetes mellitus and oxidative stress in rats by oil of Eruca sativa seeds. AB - Clinical research has confirmed the efficacy of several plant extracts in the modulation of oxidative stress associated with diabetes mellitus (DM). Oil of Eruca sativa seeds (ESS) is tried for prevention and treatment of DM induced experimentally by alloxan injection. A single dose of alloxan (100 mg/kg) produced a decrease in insulin level, hyperglycemia, elevated total lipids, triglycerides and cholesterol, decreased high-density lipoprotein and hepatic glycogen contents and elevated hepatic glucose-6-phosphatase activity. Concurrent with these changes, there was an increase in the concentration of malondialdehyde and 4-hydroxynonenal in the liver. This oxidative stress was related to a decreased glutathione (GSH) content and superoxide dismutase activity in the liver of alloxan-diabetic rats. ESS oil (0.06 ml/kg) on its own increased significantly hepatic GSH. Daily oral administration of ESS oil 2 weeks before or after diabetes induction ameliorated hyperglycemia, improved lipid profile, blunted the increase in malondialdehyde and 4-hydroxynonenal and stimulated the GSH production in the liver of alloxan-treated rats. We suggested that ESS oil could be used as antidiabetic complement in case of DM. This may be related to its antioxidative properties and to the increase in hepatic GSH. PMID- 11053895 TI - Daytime alertness, mood, psychomotor performances, and oral temperature during Ramadan intermittent fasting. AB - During the month of Ramadan, Moslems abstain from drinking and eating daily between sunrise and sunset. This change of meals schedule is accompanied with changes in sleep habit, which may affect diurnal alertness. This study examined the effect of Ramadan intermittent fasting on the diurnal alertness and oral temperature in 10 healthy young subjects. The cognitive task battery including movement reaction time (MRT), critical flicker fusion (CFF) and visual analogue scale, was administered at 6 different times of the day: 09.00, 11.00, 13.00, 16.00, 20.00 and 23.00 h on the 6th, 15th, and 28th days of Ramadan. The baseline day was scheduled one week before Ramadan, and the recovery day 18 days after this month. Oral temperature was measured prior to each test session and at 00.00 h. During Ramadan oral temperature decreased at 09.00, 11.00, 13.00, 16.00 and 20.00 h and increased at 23.00 and 00.00 h. Subjective alertness decreased at 09.00 and 16.00 h and increased at 23.00 h. Mood decreased at 16.00 h. MRT was increased at the beginning of Ramadan (R6) and CFF was not changed. These results showed that daytime oral temperature, subjective alertness and mood were decreased during Ramadan intermittent fasting. PMID- 11053896 TI - Dietary habits and cardiovascular risk in the Spanish population: the DRECE study (I). Diet and Cardiovascular Events Risk in Spain. AB - BACKGROUNDS/AIMS: To investigate dietary habits and their evolution with regard to cardiovascular risk status in Spain. METHODS: Cross-sectional study performed in two phases in 1991 and 1996 in 43 primary care clinics. One thousand and two hundred people classified as 'with cardiovascular risk' and 600 'without risk' were studied. Each participant answered a food frequency questionnaire. RESULTS: The risk group did not change oil, cereals and dairy products consumption, decreased egg, legume and meat, and increased fish, fruits and vegetables intake. The control group differed in increasing dairy products and not decreasing eggs and vegetables consumption. A small decrease in energy intake happened, from 11,315. 1 to 10,941.5 kJ in the risk group (p < 0.05). Carbohydrates intake showed a not statistically significant falling trend from 41.3 to 40.6% in people at risk and 41.8 to 40.7% in those without risk. Protein intake increased in both groups up to 16.5% and fat consumption kept at around 42.9% in both groups. The decrease in saturated fat and increase in polyunsaturated fat were statistically significant in people at risk (p = 0.000). High cholesterol intakes were found. CONCLUSION: People with cardiovascular problems changed their dietary habits in a 'healthier' way than people without risk. PMID- 11053897 TI - Changes of HDL subfractions by eicosapentaenoic acid intake and physical load in 20- to 25-year-old men. AB - The effects of daily eicosapentaenoic acid (EPA) intake and physical activity on high-density lipoprotein (HDL) subfraction, which may be an index of health status, were examined. The HDL subfraction, triglyceride and T-chol levels in the serum of 10 male volunteers aged 20-25 were examined before, immediately after and 1 h after being subjected to a physical load by bicycle ergometer at 90 W for 20 min. Subjects were then given 1.25 g EPA/day for 2 weeks, and the above test was repeated. By EPA intake, the distribution of HDL3b and 3c decreased significantly by 16.8 and 15.3%, respectively, and that of 2b increased significantly by 17.9%. The rate of change of subfraction of the 29th part (2b) of 30 parts in the total range of HDL increased by 67%, and decreased by 47% in 7th part (3c). By physical load, the distribution of HDL2a increased significantly by 15.4%, while 3b tended to decrease. By physical load after EPA intake, the distribution of 2a decreased significantly by 9.7%, and those of 3b and 3c increased significantly by 20.5 and 5.4%, respectively, and that of the 7th part (3c) increased by 37%. Thus, the physical load after EPA intake is effective to prevent arteriosclerosis as increasing the rate of change of HDL3c and as showing the longevity pattern of the HDL subfraction. Concentration of TG in a modal HDL pattern group increased by 95% after EPA intake, but that of a bimodal group did not show any change. HDL-cholesterol level in the bimodal group was higher than that in the modal group, especially after EPA intake. Two type III subjects changed to type IV by the load and the EPA intake, respectively. Thus, it seemed that the transformation from a modal pattern to a bimodal pattern by a certain lifestyle, especially regular physical activity and proper food intake, is a very important trial for the prevention of cerebrovascular diseases. PMID- 11053898 TI - Effect of increasing sucrose concentrations on oral lactic acid production. AB - Oral lactic acid production was studied on 11 healthy dental student volunteers (5 males and 6 females) during clearance of five solutions containing 5, 10, 15, 20 and 30% of sucrose. Oral fluid samples were collected at times zero, immediately before taking the sucrose solutions ('baseline'), and 15 min following intake of the solutions. The samples were analyzed qualitatively and quantitatively for the presence of lactic acid and remaining sugars using high performance liquid chromatography (HPLC). Results indicate that the amount of lactic acid in oral fluid significantly increased with increasing sucrose concentrations, up to 15% sucrose. With the higher sucrose concentrations (20 and 30%) equivalent or lower amounts of lactic acid were obtained. The threshold level for maximum lactic acid production was found to be between 10 and 15% sucrose. Statistical analysis of the data (Student's t test) indicated a significant difference in lactic acid production between the 5 and 10% sucrose solutions versus the 15, 20 and 30% sucrose solutions tested (p < 0.05). CLINICAL SIGNIFICANCE: Ingestion of solutions with higher sucrose concentrations (>15%) produced similar amounts or less of lactic acid during oral clearance than solutions containing lower sucrose concentrations (<15%). PMID- 11053899 TI - Vitamin and mineral supplements for the use of children on the German market: products, nutrients, dosages. AB - A market survey of vitamin and mineral supplements was carried out considering various points of purchase. 37 of 146 producers offered a total of 110 supplements marketed 'directly' (42%) or 'indirectly' (58%) for the use of children. These products contained 31 different nutrients, vitamins more frequently than minerals. The number of nutrients per product ranged from 1 (23%) to over 10 (13%), maximum 26. In most cases, the daily dosage recommended by producers reached from 100 to 200% of the reference doses for nutrition labelling for vitamins and from 50 to 100% for minerals. In total, the findings point to an abundance of products, nutrients and dosages in the German market supply of vitamin and mineral supplements for the use of children. PMID- 11053900 TI - Plasma copper concentration as marker of copper intake from food. AB - Copper concentration in blood plasma is used mainly as an indicator for alimentary copper supply. This has some limitations because copper concentration in plasma fluctuates with age, exercise and health status and does not increase after a meal nor decrease during short-term fasting. The Austrian Study on Nutritional Status (ASNS) provides data on nutrient intake and corresponding plasma concentrations, permitting to evaluate the suitability of copper plasma concentration as a marker of copper intake. Copper intake and intake of relevant nutrients and food groups were evaluated from 7-day weighed food records from 2,400 preschool and schoolchildren between 4 and 19 years of age. Blood samples were collected from a subgroup of 1,400 of these children. The copper concentration in plasma was measured by graphite furnace atomic absorption spectrometry. Correlations between plasma concentrations and intake of nutrients and food groups were calculated using a stepwise linear regression model. The results showed that the determination of copper plasma concentrations are inadequate to predict copper intake from dietary records. A better indicator for copper supply may be the evaluation of those food items, which are the predominant sources of copper intake at the population level. The highest influence on copper plasma concentration was found for the intake of meat, representing not only the most important food group for copper supply but also the most important food group for fat and protein supply. The individual copper status can only be estimated accurately when information on characteristic enzyme activities under control of additional factors of influence or even body stores of copper are available. PMID- 11053901 TI - Homocysteine levels in vegetarians versus omnivores. AB - Vitamin B(12), folate, and vitamin B(6) are the main determinants of homocysteinemia. The vegan diet provides no vitamin B(12), but also less strict forms of alternative nutrition may suffer from a deficit of this vitamin. The plasma homocysteine level was measured in alternative nutrition groups of adults (lacto- and lactoovovegetarians, n = 62; vegans, n = 32) and compared with the levels in a group consuming traditional diet (n = 59), omnivores). In the group of vegetarians the average homocysteine level is 13.18 vs. 10.19 micromol/l in omnivores; the frequency of hyperhomocysteinemia is 29 vs. 5% in omnivores. In the group of vegans the average homocysteine value is 15.79 micromol/l (53% of the individual values exceeded 15 micromol/l). Omnivores consume the recommended amount of methionine; however, in individuals consuming an alternative diet, the intake of methionine is deficient (assessed by food frequency questionnaire; lower content of methionine in plant proteins). Under conditions of lower methionine availability the remethylation pathway prevails; therefore, vitamin B(12) and folate were evaluated in relation to the homocysteine level. The serum vitamin B(12) levels are significantly lower in the alternative nutrition groups (214.8 pmol/l in vegetarians, 140.1 pmol/l in vegans vs. 344.7 pmol/l in omnivores); a deficit (<179.0 pmol/l) was found in 26% of the vegetarians and in 78% of the vegans vs. 0% in omnivores. The serum folate levels were within the range of reference values in all groups; however, they were significantly lower in omnivores. The results show that the mild hyperhomocysteinemia in alternative nutrition is a consequence of vitamin B(12) deficiency. PMID- 11053902 TI - In vitro activity of moxifloxacin(BAY 12-8039) against respiratory tract pathogens from six Latin-American countries. AB - The in vitro antibacterial activity of moxifloxacin (BAY 12-8039) was evaluated against 636 isolates of respiratory tract pathogens. The isolates were collected from July 1997 to August 1998 in the frame of a multinational Latin American study. E-test strips calibrated to read moxifloxacin MIC ranges from 0.002 to 32 microg/ml were used in susceptibility testing. Weekly quality control tests in each laboratory ensured reproducibility. Laboratories from Argentina, Brazil, Chile, Colombia, Mexico and Uruguay participated. MIC(90) for moxifloxacin were as follows: Streptococcus pneumoniae (304 isolates) 0.25 microg/ml, Haemophilus influenzae (135 isolates) 0.125 microg/ ml, Streptococcus pyogenes (66 isolates) 0.25 microg/ml, Moraxella catarrhalis (62 isolates) 0. 25 microg/ml and methicillin-sensitive Staphylococcus aureus (69 isolates) 0.25 microg/ml. These results agreed with reports from other areas. Moxifloxacin showed excellent activity against respiratory pathogens from participant countries. PMID- 11053903 TI - Comparison of the bactericidal activity of trovafloxacin and ciprofloxacin, alone and in combination with cefepime, against Pseudomonas aeruginosa. AB - BACKGROUND: Although ciprofloxacin exhibits more intense microbiological activity against Pseudomonas aeruginosa than does trovafloxacin, the clinical relevance of this observation remains questionable, particularly when the agents are combined with another antipseudomonal agent. METHODS: To evaluate this further, we conducted a four-way crossover trial to compare the bactericidal activities of ciprofloxacin and trovafloxacin, alone and in combination with cefepime, against three clinical isolates of P. aeruginosa. Healthy subjects received the following regimens, dosed to steady state: trovafloxacin 300 mg/24 h; ciprofloxacin 400 mg/12 h; trovafloxacin 300 mg/24 h plus cefepime 2 g/12 h, and ciprofloxacin 400 mg/12 h plus cefepime 2 g/12 h. Serum bactericidal titers were performed with each regimen. RESULTS: As monotherapy, the area under the bactericidal curve for ciprofloxacin exceeded that of trovafloxacin for all isolates. No significant difference in the overall degree of bactericidal activity was noted for two of three P. aeruginosa isolates for the combination regimens. Additionally, both combination regimens provided bactericidal activity for 100% of the dosing interval for all isolates. CONCLUSION: These results indicate that, while in vitro differences exist among these quinolones for P. aeruginosa, when a fluoroquinolone is combined with a beta-lactam, this is likely to be of little clinical significance. PMID- 11053904 TI - In vitro susceptibility to itraconazole, clotrimazole, ketoconazole and terbinafine of 100 isolates of Trichophyton rubrum. AB - BACKGROUND: A reference method for dermatophyte in vitro susceptibility testing is lacking. With the advent of new antimycotics, susceptibility testing has received increasing attention as an important laboratory tool for aiding the selection of appropriate drug therapy. METHODS: One hundred strains of Trichophyton rubrum were tested against four antifungal agents, itraconazole, clotrimazole, ketoconazole and terbinafine, by using a modification of the proposed standard M38-P of the National Committee for Clinical Laboratory Standards and two types of standardized inocula, 1.4 x 10(4) and 5 x 10(3) CFU/ml. RESULTS: Terbinafine was revealed to be the most effective antifungal drug. Of the three azole derivatives tested, clotrimazole showed the highest antifungal activity, while the minimum inhibitory concentrations (MICs) of itraconazole and ketoconazole were similar. Inoculum size did not affect the MIC of any of the antifungal agents tested. CONCLUSION: Our preliminary data provide promising results for the development of a reference method for dermatophyte susceptibility testing based on the microdilution technique, although more dermatophytes should be tested and the method evaluated in different laboratories. PMID- 11053905 TI - In vitro susceptibility of Candida dubliniensis to current and new antifungal agents. AB - BACKGROUND: Candida dubliniensis is a recently described Candida species closely related to Candida albicans, which has been associated with oral candidiasis in HIV-infected patients. Fluconazole-resistant strains of C. dubliniensis are easily obtained in vitro and this fact could be a complication if this resistance develops during treatment with this drug. METHODS: In the present study, the in vitro antifungal susceptibilities of 36 C. dubliniensis clinical isolates and culture strains to current and new antifungal agents, such as amphotericin B (AMB), amphotericin B lipid complex (ABLC), amphotericin B colloidal dispersion (ABCD), 5-fluorocytosine (5FC), fluconazole (FLC), itraconazole (ITC), ketoconazole (KTC), liposomal amphoteri- cin B (LAMB), liposomal nystatin (LNYT), LY303366 (LY), SCH56592 (SCH), and voriconazole (VRC), were determined according to the National Committee for Clinical Laboratory Standards M27-A broth microdilution method for yeasts. RESULTS: Most isolates of C. dubliniensis were susceptible to both new and current antifungal drugs, with 75.9% isolates susceptible to KTC, 86.2% to FLC and to ITC, and approximately 100% to the other antifungal agents tested. The cross-resistance phenotypes are detailed. Four isolates were resistant (MIC > or =64 microg/ml) to FLC. These 4 isolates were also resistant to KTC, and 3 of them were also resistant to ITC (MIC > or =1 microg/ml for both agents). However, these isolates were highly susceptible to 5FC and all polyene formulations (AMB, ABLC, ABCD, LAMB, and LNYT), triazole (SCH and VRC) and echinocandin (LY) antifungal agents. CONCLUSION: The new liposomal and lipidic formulations of AMB, LNYT, and the new triazoles and echinocandins may provide new alternatives to FLC for the treatment of infections by C. dubliniensis. PMID- 11053906 TI - In vitro analysis of the change in resistance of Chlamydia trachomatis under exposure to sub-MIC levofloxacin for a therapeutic term. AB - BACKGROUND: While fluoroquinolone-resistant Chlamydia trachomatis strains have not been clinically isolated, they were isolated in an in vitro study recently. METHODS: To determine whether C. trachomatis strains develop resistance under sub MIC antibacterial exposure in a clinical therapeutic term, C. trachomatis strains were exposed to sub-MIC levofloxacin (LVFX) for about 2 weeks. The MIC of LVFX was measured and DNA fingerprinting was performed every 72 h by PCR using random primers. RESULTS: There was almost no change in the MIC under exposure to 0.125 microg/ml LVFX. However, some mutational changes in DNA fingerprints developed. CONCLUSIONS: In clinical therapeutic terms, resistant strains of C. trachomatis will probably not develop, even if sub-MIC LVFX is employed. PMID- 11053907 TI - Tunicamycin treatment reduces intracellular glutathione levels: effect on the metastatic potential of the rhabdomyosarcoma cell line S4MH. AB - Highly metastatic cells are known to overexpress certain Asn-linked oligosaccharides in the plasmatic membrane. Another phenotypic characteristic of malignant cells consists in the expression of high levels of intracellular glutathione (GSH). The aim of the present work was to demonstrate that the inhibition of N-glycosylation induces changes in intracellular GSH levels, and in turn participates in the inhibition of the metastatic potential of tumor cells by tunicamycin treatment. Firstly, we demonstrated that in comparison to the poorly metastatic cell line F21, the highly metastatic cells S4MH express a higher number of Asn-linked beta1-6 branched oligosaccharides and sialic acid (SA) and/or chitobiose oligosaccharides in glycoproteins involved in the regulation of the adhesion efficiency of tumor cells on endothelial cells and extracellular matrix. Our results showed that the decrease in S4MH cell adhesion efficiency on endothelial cells and extracellular matrix after the inhibition of N-glycan processing by tunicamycin treatment was caused by: (1) inhibition of the expression of N-glycan structures recognized by endothelial endogenous lectins, including beta1-6 branched oligosaccharides and SA and/or chitobiose oligosaccharides, and (2) redistribution of cell surface glycoproteins with beta1 6 branched oligosaccharides and/or SA and/or chitobiose oligosaccharides in their structures, caused by the depletion of intracellular GSH levels. The latter condition prevents the organization of these glycoproteins in the plasmatic membrane of S4MH cells necessary for anchoring to the substratum. PMID- 11053908 TI - Indomethacin inhibits the accumulation of tumor cells in mouse lungs and subsequent growth of lung metastases. AB - BACKGROUND: The interaction of cancer cells with blood cells and cell wall components evokes inflammatory responses and is a critical event in the metastatic process. Indomethacin is a potent inhibitor of the cyclooxygenase (COX) enzymes and has previously been shown to decrease the growth of primary tumors in vivo. Proinflammatory prostaglandins produced by the two COX enzymes may also play a role in the development of metastases. METHODS: To directly address this question, we tested the effect of indomethacin on the accumulation of circulating [(3)H]-uridine-labeled tumor cells in the lungs and on the subsequent development of lung tumors. RESULTS: We found that inhibition of COX activity in the recipient mice prior to the injection of tumor cells decreased the percentage of the cells arrested in the lungs. This effect was highly significant since it subsequently led to substantial attenuation of lung metastasis development. CONCLUSION: These data thus demonstrate the antimetastatic effect of indomethacin through a mechanism which involves a reduction in tumor cell uptake by the lungs. PMID- 11053909 TI - A randomized prospective study of oral versus intravenous antibiotic prophylaxis against postoperative infection after sagittal split ramus osteotomy of the mandible. AB - BACKGROUND: We investigated whether oral levofloxacin was as effective as intravenous cefazolin sodium for preventing postoperative infections in patients undergoing sagittal split ramus osteotomy of the mandible. METHODS: Forty-four patients were randomized to treatment with levofloxacin or cefazolin sodium. Levofloxacin (100 mg t.d.s.) was administered orally, and cefazolin sodium (1 g b.i.d.) by intravenous infusion. Both drugs were given until 5 days postoperatively. The concentrations of levofloxacin in the mandibular bone marrow and serum were measured. RESULTS: The mean levofloxacin concentration 4 h and 53.7 min after administration was 1.086 microg/ml in serum and 1.328 microg/ml in the mandibular bone marrow. The efficacy rate of levofloxacin was 95% and that of cefazolin sodium was 85.7%, with no significant difference between the two drugs (p = 0.322). CONCLUSION: Oral administration of levofloxacin is a simple, cost effective and safe alternative to intravenous antibiotic prophylaxis after mandibular surgery. PMID- 11053910 TI - Matting of hair: a multifactorial enigma. AB - For matting of hair, multifactorial mechanisms are responsible. Thereby physical conditions, chemical agents and behavioural factors play the main roles. It seems evident that hair matting is not a hair disease; consequently, cutting off the affected hair and avoiding all possible trigger factors are the therapy of choice. PMID- 11053911 TI - Cell-kinetic evidence for increased recruitment of cycling epidermal cells in psoriasis: the ratio of histone and Ki-67 antigen expression is constant. AB - BACKGROUND: One of the hallmarks of the psoriatic plaque is increased epidermal proliferation. Whether this is the result of an increased recruitment of cycling epidermal cells or a decrease in cell cycle time has been a matter of debate for years. OBJECTIVE: Calculating cell-kinetic information from the number of S phase cells in psoriasis by in situ hybridisation using a histone probe and the number of cycling epidermal cells by immunohistochemistry using the MIB-1 antibody. METHODS: Immunohistochemistry and non-isotopic in situ hybridisation were performed on serial sections of 33 untreated psoriatic samples and 14 tacalcitol treated samples. RESULTS: The labelling index (number of cells in S phase/number cycling cells per millimetre length of section) in psoriatic untreated as well as in treated plaques is 16%. The amount of S phase cells in our experiment is equal compared with the number of cells in S phase as determined by BrdU incorporation. CONCLUSION: Using this direct approach to study cell-kinetic behaviour of psoriatic skin, we reconfirm that the psoriatic abnormality is due to a defect in the G(0)-G(1) recruitment mechanism (by increased recruitment of G(0) cells), a decrease in apoptosis or an increase in the number of cell divisions in the transit-amplifying compartment, rather than a reduction in the cell cycle time. PMID- 11053912 TI - Investigation of annexin V binding to lymphocytes after extracorporeal photoimmunotherapy as an early marker of apoptosis. AB - BACKGROUND: Induction of programmed cell death is assumed to be a possible effect of extracorporeal photoimmunotherapy (ECPI). OBJECTIVE: In the present study lymphocytes of patients with cutaneous T cell lymphoma undergoing ECPI were investigated for early apoptotic events. METHODS: Annexin V, known for its selective affinity to phospholipids, was used to detect early phases of apoptosis. Simultaneous staining with propidium iodide binding to DNA allowed detection of late apoptotic/necrotic cells. RESULTS: At 1 h after ECPI, an increase in early apoptotic cells was found indicating a direct effect of ECPI. At 20 h after each ECPI session, a delayed increase in the number of apoptotic lymphocytes was observed in early apoptotic annexin-stained cells and in late apoptotic cells, whereas in nonirradiated cells no remarkable changes were found. Apoptosis was confirmed by altered light scattering properties and DNA fragmentation. CONCLUSION: The apoptotic cell death of reinfused lymphocytes is supposed to be a therapeutic effect of ECPI. PMID- 11053913 TI - Unraveling the patterns of subclinical pheomelanin-enriched facial hyperpigmentation: effect of depigmenting agents. AB - BACKGROUND: During photoaging, the density of melanin chromatophores is heterogeneous in the epidermis. AIMS: To define the patterns of pheomelanin enriched melanotic hypermelanosis of the face in phototype II subjects and to assess the effect of depigmenting agents. Azelaic acid and glycolic acid were tested as well as a soy extract, reported to reduce pigmentation through interaction with the protease-activated receptor 2 (PAR-2) of keratinocytes. METHOD: Evaluations were made by image analysis of high magnification pictures obtained by a video camera equipped with an internal ultraviolet-emitting unit (Visioscan((R))). RESULTS: Three patterns of subclinical facial hypermelanosis were recognized including the spotty perifollicular type, the accretive globular type and the elongated type of the sunny side of wrinkles. Azelaic acid and the soy extract led to significant skin lightening after a 3-week treatment. By contrast, glycolic acid showed an inconsistent effect. CONCLUSION: Sensitive fluorescence video recording combined with image analysis represents an advance in the noninvasive assessment of the mottled subclinical skin pigmentation. The depigmenting effect observed with the soy extract indicates that the inhibition of PAR-2 may be a novel way to approach certain pigmentary disorders of the skin. PMID- 11053914 TI - Differentiation between merkel cell carcinoma and malignant melanoma: An immunohistochemical study. AB - BACKGROUND: Although Merkel cell carcinoma (MCC) exhibits specific clinical and histologic features, differentiation from other cutaneous neoplasms, such as lymphoma, metastatic oat cell carcinoma and malignant melanoma (MM), may sometimes be difficult. OBJECTIVE: The aim of our study was to immunohistochemically differentiate MCC from MM. METHODS: Paraffin sections from 6 cases of primary MCC and 6 cases of primary MM were investigated. For immunostaining, the APAAP method was used. RESULTS: Neuron-specific enolase was positive in all cases of MCC, as well as in 2 cases of MM. Marked positivity for cytokeratins 18, 20 and chromogranin A was observed in the MCC group, whereas a complete absence of expression of these three markers was noted in the MM group. Immunostaining with HMB45 and NKI/C3 was positive in all cases of MM and negative in all cases of MCC. S-100 protein was positive in all but 1 case of MM. In contrast, only 1 case of MCC reacted with S-100 protein. CONCLUSION: Our results underline the role of immunohistochemistry in the diagnosis and differential diagnosis of MCC. In particular, the combination of neuron-specific enolase, cytokeratins 18, 20 and chromogranin A positivity for MCC and HMB45, NKI/C3 and S 100 protein positivity for MM is of great value in the distinction between these two cutaneous neoplasms. PMID- 11053915 TI - Contact urticaria from latex in healthcare workers. AB - BACKGROUND: Latex allergy is an important medical problem for an increasing number of patients. It has been documented as causing immediate hypersensitivity reactions ranging from mild urticaria to life-threatening anaphylaxis after cutaneous, mucosal or visceral exposure. Recent studies in northern Europe and the USA suggest that between 2.8 and 16.9% of healthcare workers are affected by latex hypersensitivity type I reactions. OBJECTIVES: To test the prevalence of contact urticaria from latex gloves in a group of healthcare workers, to examine the factors associated with latex allergy and to evaluate some diagnostic methods used in latex allergy. METHODS: A total of 929 employees of the surgical units who used latex gloves on a regular basis, at least once a day, were invited to participate in this study including administration of a questionnaire, a prick test with a commercial extract of latex, a prick test with latex glove eluate, a use test, RAST and an immunoblotting system; moreover, a prick test with a group of common inhalant allergens and a prick-by-prick test with fresh fruit (banana, kiwi, avocado, chestnut) were employed. RESULTS: Of the 929 staff sent questionnaires, 313 (33.5%) replied; of those who responded, 118 gave a history of hand problems such as itch, erythema, wheals when wearing gloves, dryness and irritation most marked on the backs of the hands. Among these 118 workers, 16 refused skin testing and examination of blood, so 102 subjects were studied for latex allergy; 21/118 (17.8%) healthcare workers were found to be latex allergic. Eighty-one staff members gave a history of hand problems worsened by wearing gloves but were not latex allergic on testing. Those healthcare workers who completed the questionnaire and answered negatively (195/313) were not tested for latex allergy. Prick tests with the commercial solution were positive in 11 of the 21 subjects studied; prick tests with the eluate of glove, RAST and the use test were positive in all workers; 10 of the 21 sera showed positive immunoblot results. Atopy and a preexisting irritant contact eczema of the hands were present in a high percentage of the workers. CONCLUSION: In this study of healthcare personnel, we found that allergic contact urticaria from latex was present in 21 workers of the 313 (6.7%) who responded to the questionnaire and of the 102 (20.5%) who were tested for latex allergy. Atopy and irritant contact eczema of the hands were frequent in these subjects. Skin prick testing with latex glove eluate and the use test seem to be more sensitive than in vitro testing, particularly immunoblotting, and are biologically more relevant; skin testing with glove eluate must be preferred to testing with a commercial extract. PMID- 11053916 TI - Pemphigus foliaceus induced by exposure to sunlight. Report of a case and analysis of photochallenge-induced lesions. AB - BACKGROUND: Although there are many reports on photo-induced pemphigus, careful analysis in the development of acantholytic blister have rarely been performed in the lesions induced by photochallenge. OBJECTIVE: The study was intended to elucidate the mechanisms by which ultraviolet light (UV) radiation causes the skin lesions of pemphigus foliaceus. METHODS: Photochallenge-induced lesions were examined histologically and immunohistochemically over the time course with three biopsy specimens. Neutrophil adhesion assay was performed using the specimens prepared from the photochallenged lesions as substrates. RESULTS: A small number of neutrophils were seen at 5 h after photochallenge in the dermis and epidermis, and slight acantholysis was detected at 72 h. In neutrophil adhesion assay, greater numbers of neutrophils adhered to the upper epidermis of the skin obtained at 5 h after challenge. CONCLUSIONS: Our results suggest that both enhanced binding of autoantibodies to the epidermis after UV radiation and preferential adhesion of neutrophils to the UV-irradiated epidermis contribute to the development of acantholysis in photo- induced pemphigus foliaceus. Intercellular adhesion molecule-1 (ICAM-1) expression on keratinocytes is not primarily responsible for the epidermal migration of neutrophils. PMID- 11053917 TI - The efficacy, safety and tolerance of calcitriol 3 microg/g ointment in the treatment of plaque psoriasis: a comparison with short-contact dithranol. AB - OBJECTIVE: A comparison of efficacy, safety and tolerance of a twice-daily application of calcitriol 3 microg/g ointment with dithranol cream. METHODS: The study was an 8-week, prospective, randomised, open, parallel-group trial with 114 patients. Subjects received either 3 microg/g calcitriol ointment (twice daily) or 0. 25-2% dithranol cream (once daily for 30 min). Results were measured using global improvement, global severity, PASI, quality of life (QOL) and Psoriasis Disability Index scores. Safety was determined from reports of adverse events and blood chemistry analysis. RESULTS: At final assessment, calcitriol and dithranol were comparably efficacious. Skin irritation was reported by 5% of calcitriol and 72% of dithranol patients. Patients rated QOL and overall acceptability of calcitriol more highly. CONCLUSIONS: Twice-daily calcitriol ointment (3 microg/g) is equally effective as a once-daily, short-contact dithranol regimen. However, calcitriol is better tolerated than dithranol, and provides a better QOL and a greater patient acceptability. PMID- 11053918 TI - High prevalence of seborrhoeic dermatitis on the face and scalp in mountain guides. AB - BACKGROUND: High incidence rates of seborrhoeic dermatitis (SD) have been reported in HIV-infected individuals, indicating immunosuppression to be involved in the pathogenesis. OBJECTIVE: To establish the prevalence of SD in mountain guides who have a high occupational exposure to solar UV radiation. PATIENTS AND METHODS: In November 1999, 283 mountain guides were physically examined on the face and scalp for symptoms of SD in Austria (n = 75), Switzerland (n = 123) and Germany (n = 85); they were 21.3-93.1 years of age (median age 41.4 years). RESULTS: Forty-six of 283 (16. 3%) mountain guides when examined clinically were found to have SD. The median age of mountain guides with SD was 41.2 years. There were similar incidence rates in all three countries. CONCLUSION: SD affects mountain guides in a clearly higher percentage as the general population. We suggest UV-induced immunosuppression due to occupational sun exposure as a pathogenetic factor. PMID- 11053919 TI - Multifocal fixed drug eruption to paracetamol, tropisetron and ondansetron induced by interleukin 2. AB - Fixed drug eruptions (FDEs) represent an uncommon subset of drug reactions where typically dusky red skin eruptions recur at the same site each time a drug is administered. Multifocal FDEs are defined by skin eruptions at more than one site. We describe a patient with metastatic melanoma who developed a multifocal FDE to paracetamol, tropisetron and ondansetron during chemoimmunotherapy with interleukin 2 (IL-2) and dacarbazine. This case is unique since to our knowledge there has been no previous report of such a drug reaction to tropisetron. Moreover, recurrences were induced by drugs which were chemically unrelated (i.e. tropisetron and paracetamol). We propose that this unusual skin reaction to multiple drugs was induced by IL-2 administered during immunochemotherapy for the metastatic melanoma. PMID- 11053920 TI - A case of pencil core granuloma with an unusual temporal profile. AB - The patient is a 47-year-old female with a bluish tumor resembling malignant melanoma at the macroscopic level on the medial aspect of her left big toe. The patient had crushed a pencil with this toe about 30 years previously and, since then, a bluish lesion had been present. About 15 years ago, the lesion had suddenly increased in size over the course of a few months. Subsequently, the size of the lesion had not changed noticeably. Ultrasonography, but not magnetic resonance imaging was a helpful preoperative examination to distinguish the lesion from malignant melanoma. The excised lesion contained a piece of material that resembled pencil lead and bluish mud. X-ray microanalysis of the lead-like material revealed that its composition was similar to that of pencil lead. Histologic examination showed features of foreign-body reaction, except for necrotic change and few histiocytes. PMID- 11053921 TI - Cutaneous ulceration after injection of polyethylene-glycol-modified interferon alpha associated with visual disturbances in a melanoma patient. AB - Interferons are used in the therapy of multiple sclerosis, Kaposi's sarcoma, hepatitis and melanoma. Their short half-life that requires frequent injections can be increased by polyethylene glycol (PEG) modification. A 50-year-old patient was diagnosed as having an acrolentiginous melanoma (Breslow >5 mm, Clark level IV) and inguinal lymph node metastases. After surgical excision and lymphadenectomy, immune therapy with 6.0 microg pegylated interferon alpha(2b)/kg body weight, s.c., was started. Cutaneous ulcerations at the injection sites developed 9 months after treatment initiation. The patient also developed blurred vision and presented with binasal scotomas and pathological visually evoked potentials and electroretinogram. The cutaneous ulcerations slowly healed under local therapy and reduction of the concentration of the PEG-modified interferon from 0.86 to 0.43 mg/ml. The dosage was maintained. Two months later, the therapy was stopped due to disease progression. Vision subsequently recovered. Cutaneous reactions evolved at the sites of subcutaneous injections of PEG-modified interferon alpha(2b). Changes in vision can probably be attributed to immunotherapy. PMID- 11053922 TI - Lack of TP53 mutations in a case of porokeratosis palmaris, plantaris et disseminata. AB - A 73-year-old man with porokeratosis palmaris, plantaris et disseminata is presented. He had punctate, guttate and annular hyperkeratotic papular lesions widespread on his body with thorn-like hyperkeratosis on the palms and soles. Lesional skin did not show mutations of TP53 exons 5-6, 7, 8. PMID- 11053923 TI - Two cases of Kimura's disease with unusual clinical manifestations. AB - We report 2 cases of Kimura's disease with unusual clinical presentation. Lesions were itching nodules located at the elbow and the dorsum of the hand, and were associated with unusual laboratory values. These features may lead to problems in clinical differential diagnosis. PMID- 11053924 TI - Carpal tunnel syndrome with ulcerous skin lesions. AB - We report on a patient suffering from an advanced stage of carpal tunnel syndrome (CTS) with entire loss of cutaneous sensibility and impaired motor function who developed ulcerous skin lesions. In the present case, we would like to focus the attention of the dermatologist on CTS, when skin changes, i.e. atrophy, anhidrosis and ulcerations are found in the sensory zones of the median nerve. The necessity to explore a potential trauma is stressed. PMID- 11053925 TI - A case of lipedematous alopecia occurring in a male patient. AB - A 30-year-old Japanese man presented with a thickening of the scalp and diffuse alopecia on the vertex. A skin biopsy specimen of the thickened scalp showed an increased thickness of subcutaneous adipose tissue and a marked decrease in the number of hair follicles. By an MRI examination, the irregularly thickened subcutaneous fatty tissue was visualized. These findings indicated that the patient had been suffering from lipedematous alopecia. This is a rare condition characterized by diffuse alopecia due to the thickening of the layer of adipose tissue covering the scalp. Only 4 female cases of lipedematous alopecia have been reported previously. MRI is thought to be useful for the differentiation of lipedematous alopecia from lipoma. The present case is considered to present the first male patient with lipedematous alopecia. PMID- 11053926 TI - Prolonged photosensitivity following contact photoallergy to ketoprofen. AB - We report the third case of prolonged photosensitivity secondary to contact photoallergy to topical ketoprofen, a 2-arylpropionic acid derivative. The patient suffered from persistent photosensitivity for more than 1 year after the withdrawal of ketoprofen with recurrent eruptions on sun-exposed skin areas. This photosensitivity was associated with a persistent decrease in polychromatic and UVA minimal erythemal doses. Photobiological testing revealed cross-reactivity with fenofibrate and benzophenones. Photoallergy to ketoprofen is due to the benzophenone structure or to the very similar thiophene phenylketone of tiaprofenic acid, but not to the arylpropionic function. Thus, fenofibrate, tiaprofenic acid and benzophenones should be avoided by patients with a positive history of photocontact dermatitis to ketoprofen. PMID- 11053927 TI - Successful treatment of discoid lupus erythematosus with argon laser. AB - Vascular lesions with telangiectasias on visible areas, such as the face, are common in discoid lupus erythematosus (DLE); however, an efficient management of these skin lesions can sometimes be difficult. Since argon laser light is able to specifically coagulate vascular structures, it has been used in the treatment of various vascular skin malformations. Therefore, we addressed the issue whether argon laser treatment could be a therapeutic alternative for this disease. Here, we report on a patient with DLE, who suffered from long-standing erythematous, telangiectatic plaques on the face refractory to standard regimens of therapy. After 2 laser applications, a significant improvement was observed and after 5 sessions of argon laser therapy the treated skin lesions had completely resolved with an excellent cosmetic result. The patient tolerated the laser treatment well without any short-term side effects. These data indicate that argon laser therapy might be a powerful alternative approach in the treatment of vascular skin lesions of DLE. PMID- 11053928 TI - Extramammary Paget's disease in siblings. PMID- 11053929 TI - Treatment of plantar hyperhidrosis with dermojet injections of botulinum toxin. PMID- 11053930 TI - Psoriasis associated with HCV and exacerbated by interferon alpha: complete clearance with acitretin during interferon alpha treatment for chronic active hepatitis. PMID- 11053931 TI - Multiple anemic macules on the arms: a variant form of nevus anemicus? PMID- 11053932 TI - How unstable is the concept of stability in surgical repigmentation of vitiligo? PMID- 11053933 TI - Leucoderma associated with the use of topical minoxidil: a report of two cases. PMID- 11053934 TI - 'Plica neuropathica': matting of hair. PMID- 11053935 TI - Rheumatoid neutrophilic dermatitis/sweet's syndrome in a patient with seronegative rheumatoid arthritis. PMID- 11053936 TI - Mesenteric cysts. Toward less confusion? AB - OBJECTIVE: Mesenteric cysts are rare intra-abdominal masses generally omited or briefly reported in textbooks. Their rarity had fostered a lack of information and difficulty in classification. The goal of the study is to present a simple, comprehensive, and reproducible classification of mesenteric cysts. METHODS: A large review of the literature with particular attention to recent series was performed in order to identify and characterize the different groups of mesenteric cysts. RESULTS: Although cysts of lymphatic and mesothelial origin are those most frequently encountered, they were confounded or omitted in previous classifications. However, their distinction is important because their incidence and mode of presentation differ. Lymphangiomas predominate in male children, may cause acute abdominal pain, and frequently require resection of adjacent structures. In contrast, cysts of mesothelial origin such as benign cystic mesothelioma are seen more frequently in young and middle-aged women, usually present with more indolent symptoms, and often recurs. CONCLUSIONS: A new classification based essentially on histopathological features should include the 6 following groups: (1) cysts of lymphatic origin (simple lymphatic cyst and lymphangioma); (2) cysts of mesothelial origin (simple mesothelial cyst, benign cystic mesothelioma, and malignant cystic mesothelioma); (3) cysts of enteric origin (enteric cyst and enteric duplication cyst); (4) cysts of urogenital origin; (5) mature cystic teratoma (dermoid cysts), and (6) pseudocysts (infectious and traumatic cysts). PMID- 11053937 TI - Two-step selective clamping of IVC for removal of hepatocellular carcinoma with intracaval extension. AB - Hepatocellular carcinoma (HCC) with retrohepatic intracaval extensions are difficult to treat. HCC may sometimes extend into the inferior vena cava (IVC) through two routes: via the right hepatic vein and via the inferior right hepatic vein. In such cases, in which tumor emboli are located both above and below the confluence of the hepatic vein with the IVC, we first remove the upper embolus during THVE, and then remove the lower one while the IVC is clamped obliquely in order to preserve the residual liver circulation. PMID- 11053938 TI - Short-term hemodynamic effects of transjugular retrograde obliteration of gastric varices with gastrorenal shunt. AB - OBJECTIVES: The purpose of this study was to investigate the short-term effects on portal hemodynamics of transjugular retrograde obliteration (TJO) of gastric varices with gastrorenal shunt. METHODS: Thirty patients with gastric varices and a gastrorenal shunt were included in this study. The patients ranged in age from 42 to 75 years (16 men and 14 women), and according to Child's classification, class A, B and C cirrhosis was seen in 1, 13 and 16 patients, respectively. The portal blood flow was measured by an ultrasonic duplex Doppler system, and the wedged hepatic venous pressure was measured by hepatic venous catheterization, before and after TJO. RESULTS: Complete obliteration of the gastrorenal shunt and gastric varices was revealed by retrograde inferior phrenic venography and computed tomography after TJO in all cases. The wedged hepatic venous pressure was significantly increased the day after TJO compared with that before therapy (257 +/- 71 vs. 307 +/- 73 mm H(2)O, p < 0.01). The portal venous flow was significantly increased 1 week after TJO compared with that before therapy (744 +/- 190 vs. 946 +/- 166 ml/min, p < 0.01). The serum albumin levels before and after TJO were 3.0 +/- 0.4 and 3.1 +/- 0.5 g/dl, respectively, and the total bilirubin levels were 1.5 +/- 0.7 and 1. 5 +/- 0.8 mg/dl, respectively, neither of these parameters changing significantly. The plasma ammonia levels before and after TJO were 109 +/- 62 and 67 +/- 31 microg/dl, and the indocyanine green retention rates at 15 min were 31 +/- 13 and 24 +/- 13%, both showing a significant change (p < 0.01 and p < 0.05, respectively). CONCLUSIONS: We conclude that TJO increases portal blood flow which contributes to the decrease in plasma ammonia levels and the indocyanine green retention rate, although increasing the wedged hepatic venous pressure. PMID- 11053939 TI - Effects of perioperative protease inhibitor on inflammatory cytokines and acute phase proteins in patients with hepatic resection. AB - AIM: The objective of this study was to examine the effects of perioperative administration of ulinastatin, or urinary trypsin inhibitor (UTI), on inflammatory cytokines and acute-phase proteins induced by inflammatory cytokines in patients who had undergone hepatic resection. METHOD: Twenty patients admitted to the hospital for hepatic resection were equally randomized to one of two groups: the UTI group, those who were administered perioperative UTI, and the control group. RESULTS: The UTI group had no adverse effects from using UTI. Production of serum interleukin-6 (IL-6) tended to be attenuated in the UTI group when compared with the control group. Moreover, the UTI group had significantly decreased positive acute-phase C-reactive protein (p < 0.05) and significantly increased negative acute-phase protein prealbumin and retinol-binding protein (p < 0.05). Serum IL-6 levels significantly correlated with serum C-reactive protein levels on postoperative day 1 (r = 0.70, p < 0.01). CONCLUSION: These results suggest that perioperative administration of UTI might deserve further assessment for use in modulating acute-phase responses without adverse effects in patients who have undergone hepatic resection. PMID- 11053940 TI - Laparoscopic cholecystectomy to treat patients with asymptomatic gallstones. AB - BACKGROUND/AIMS: The management of patients with silent or asymptomatic gallstones remains controversial. The objective of this study is to determine the complications of laparoscopic cholecystectomy performed to treat patients under 50 years of age with asymptomatic gallstones. METHOD: 207 patients, 13-49 years of age with asymptomatic gallstones, were subjected to laparoscopic cholecystectomy. There were 161 (78%) women and 46 (22%) men. RESULTS: Operative time varied from 25 to 95 min, with an average of 42 min. The length of hospitalization varied from 12 h to 2 days. In no patient was the operation converted to laparotomy. There was neither mortality nor major complications. The following minor complications were observed: vomiting in 43 patients (20.7%), fever in 16 (7.7%), minor skin infection in 7 (3.3%), and keloid in 2 (0. 9%). CONCLUSION: It is concluded from this study that laparoscopic cholecystectomy is a safe procedure in the treatment of patients under 50 years of age with asymptomatic gallstones. PMID- 11053941 TI - Drainage of the common bile duct by the axial transomphalic extraperitoneal route. AB - AIM: A retrospective analysis was made of results obtained in 169 patients with axial transomphalic external biliary drainage, operated between 1984 and 1998. However, the authors' experience with this method covers a total of 773 cases in which this type of drainage was used between 1966 and 1998. METHOD: The technique is described in detail. The use of the omphalic ligament provides a completely extraperitoneal trajectory for biliary drainage tubing inserted between the two peritoneal layers of the ligament. RESULTS: Postoperative mortality and morbidity in the patients presented here were not related to the drainage procedure, but were related to the background illness, associated disturbances and the surgical procedures applied. Axial transomphalic biliary drainage has many indisputable advantages in comparison with other types of external biliary drainage. Indications and counterindications of the method are summarized. CONCLUSION: The technique is especially valuable because it protects biliary-digestive anastomoses and common biliary duct sutures, facilitating implantation of prostheses and reconstruction of the main biliary pathway, as well as benign and malignant stenoses. PMID- 11053942 TI - Splenic abscess. An old disease with new interest. AB - BACKGROUND/AIMS: To study the demographics, signs and symptoms, causes, risk factors, imaging findings, bacteriologic profile, treatment and outcome of patients with splenic abscess. METHOD: The medical records of 17 patients with splenic abscess at two tertiary-care hospitals between 1989 and 1997 were retrospectively reviewed. The demographic data, physical and radiological findings, treatment, bacteriology reports and outcome of treatment were reviewed. RESULTS: The mean age of patients was 43 years (range 7-79 years). Fever and abdominal pain were the most prominent signs. Seven patients were immunocompromised, three had abscessed hydatic cysts, two were drug users and three suffered from splenic trauma, infarction, and endocarditis, respectively. No predisposing factor was identified in 2 patients. In all cases, CT demonstrated the splenic lesion(s). Staphylococcus species and Bacteriodes were the most common microbes, identified in the blood and abscess cultures. Thirteen patients underwent splenectomy, two medical therapy and two no therapy with respective survival rates of 92, 100 and 0%. CONCLUSION: Splenic abscess is a rare surgical entity encountered mostly in immunocompromised patients. CT scan is the gold standard for the definite diagnosis. Splenectomy is the treatment of choice, while medical therapy should be reserved for unusual pathogens provided that an effective antimicrobial agent is available. PMID- 11053943 TI - Ampullary but not periampullary duodenal diverticula are an etiologic factor for chronic pancreatitis. AB - BACKGROUND: Duodenal diverticula are common incidental findings. Although a definite treatment is rarely required, an association with biliary and pancreatic diseases is often suggested. Our aim was to determine the frequency of complications in relation to the location of the diverticulum. METHODS: We reviewed 64 patients with extraluminal duodenal diverticula. In 24 of these patients the diverticulum was treated surgically. The incidence of symptoms and complications is analyzed and follow-up in 88% of operated patients is presented. RESULTS: Thirty-one patients presented with ampullary, 36 patients with periampullary diverticula. Three patients had both types of diverticula. The indication for surgery in 24 patients was chronic pancreatitis (n = 12), chronic pain (n = 6), diverticular perforation (n = 3), bleeding (n = 2), or jaundice (n = 1). In 7 out of 31 patients with ampullary and none out of 36 patients with periampullary diverticula, chronic pancreatitis was considered to be induced by the diverticulum. Morbidity in 24 operated patients was 17%, no patient died. At follow-up all patients were free of symptoms. CONCLUSION: Extraluminal duodenal diverticula are frequently found. They rarely cause symptoms and need no surgical treatment. While ampullary duodenal diverticula can cause chronic pancreatitis, periampullary duodenal diverticula are no etiologic factor. PMID- 11053944 TI - The history is important in patients with suspected acute appendicitis. AB - BACKGROUND/AIMS: The clinical diagnosis of acute appendicitis is incorrect in 20 30% of patients undergoing surgery. We analysed the clinical importance of nine commonly used symptoms and signs in 544 consecutive patients with regard to the correct diagnosis of acute appendicitis. METHODS: Open population-based prospective study. The degree of the surgeon's certainty of the preoperative diagnosis was assessed. The final diagnosis was based on histology. Logistic regression was used to analyze the independent value of nine symptoms and signs to predict acute appendicitis by calculating odds ratio (OR) with 95% confidence intervals (CI). RESULTS: In 434 of 544 patients (80%) acute appendicitis was confirmed. A history of nausea or vomiting (OR = 2.3; CI = 1.11 to 4.76) and pain migration to right iliac fossa (OR = 1.9; CI = 1.12 to 3.22) were significant predictors of acute appendicitis. Pain migration was found to be an independent predictor in females and nausea or vomiting in males. In the group of patients (29%) with an uncertain preoperative diagnosis, pain migration predicted a correct diagnosis in females (OR = 4.7; CI = 1.2 to 18), while tenderness over McBurney's point was a significant predictor in males (OR = 8.3; CI = 1.1 to 63). CONCLUSIONS: A history of pain migration and nausea or vomiting were independent predictors for the correct diagnosis of acute appendicitis in patients undergoing surgery. Thus, patient history is important in this patient group. PMID- 11053945 TI - Prospective multicenter study of antibiotic prophylaxis in operative treatment of appendicitis. AB - BACKGROUND/AIMS: A prospective, multicenter observation study was conducted to investigate the effectiveness of antibiotic prophylaxis in the operative treatment of appendicitis. METHODS: Between June 1996 and May 1997, a total of 4,968 patients underwent an operation for appendicitis at 34 East German hospitals. 41.4% (n = 2,424) received perioperative antibiotic prophylaxis whereas in 58.9% of the cases this was not given. The patients who received antibiotic prophylaxis constituted a negative selection, both with regard to risk factors and to the stage of inflammation. RESULTS: In the total test group, the rate for septic disorders of wound healing amounted to 2.5% (n = 120). The use of antibiotic prophylaxis makes it possible to significant lower the incidence of postoperative septic disorders of wound healing (p < 0.001). In both conventional (0.7% with prophylaxis vs. 3.8% without) and in laparoscopic appendectomy (0 vs. 1.0%) the rate for septic disorders of wound healing can be reduced (p < 0.001). The global comparison irrespective of possible antibiotic prophylaxis shows a significant advantage for appendectomies which started with laparoscopy (p = 0.008), but in the subgroups with prophylaxis this advantage is completely neutralized (p = 0.78). CONCLUSION: From this one can deduce that conventional appendectomy with antibiotic prophylaxis comprises no higher risk of wound infection than laparoscopy, since the risk of conversion exists for every operation begun by laparoscopy. These results would lead to the conclusion that antibiotic prophylaxis should be given before every appendectomy, whether by laparoscopy or conventional methods. PMID- 11053946 TI - Effects of N-acetylcysteine on pulmonary macrophage activity after intestinal ischemia and reperfusion in rats / with invited commentaries. AB - BACKGROUND/AIMS: Intestinal ischemia and reperfusion (I/R) is considered to be a critical and triggering event in the development of distal organ dysfunction after a variety of insults. It appears that activated leukocytes, especially polymorphonuclear granulocytes (PMNs), and reactive oxygen species are important mediators in the process. In the present study, the aim was to evaluate the behavior of pulmonary macrophages, acute lung injury and pulmonary endothelial permeability after intestinal I/R, together with potential alterations in pulmonary endothelial and epithelial ultrastructure and cellular membrane system integrity. METHODS: Intestinal ischemia for 40 min was followed by reperfusion for 12 h in the rat. Macrophage uptake of radiolabeled bacteria, levels of pulmonary blood content assessed by radiolabeled red blood cells and pulmonary endothelial permeability of radiolabeled albumin, as well as pulmonary endothelial and epithelial ultrastructure and cellular membrane system integrity by the use of scanning electron microscopy and a tracer was evaluated after 12 h reperfusion. Treatment with the free radical scavenger N-acetylcysteine (NAC) administered prior to reperfusion was evaluated. RESULTS: Overactivation of pulmonary macrophages was noted after intestinal I/R, as was a significant decrease in pulmonary blood content. No increase in pulmonary albumin leakage or increase in pulmonary water content was found after intestinal I/R as compared to controls. Treatment with NAC prevented against intestinal I/R-induced overactivation of pulmonary macrophages and a decrease in pulmonary blood content. CONCLUSION: Reactive oxygen species may be involved in the regulation of pulmonary macrophage function and pulmonary circulation after intestinal I/R. PMID- 11053947 TI - Long-term results of anterior anal sphincter repair for fecal incontinence due to obstetric injury / with invited commentaries. AB - AIMS: This study is to report short- and long-term results of anterior sphincter repair for fecal incontinence due to obstetric injury and factors predicting an unsuccessful outcome. METHODS: Thirty-nine consecutive patients, mean age 51 years (range 29-74), who underwent anterior sphincter repair for fecal incontinence due to obstetric injury were investigated. Duration of symptoms ranged from 9 months to 34 years. All patients underwent an anterior overlapping sphincter muscle reconstruction and in most cases a puborectal muscle plasty. RESULTS: Three months after surgery 77% of the patients had regained continence (Parks score of 1 or 2), at 9 months 67% were continent and after 12 months or more (mean, range 12-114) only 62%. Patients with prolonged pudendal latency (>2.2 ms) did significantly worse than patients without it (p < 0.05). Patients who had had lateral episiotomy during labor had significantly better outcome than those without it (p < 0.05). CONCLUSION: The outcome of anterior sphincter repair deteriorates with time after surgery. Assessment should be done at least 1 year after surgery to evaluate the final results of anterior sphincter repair. Prolonged pudendal latency predicts a poor outcome of anterior sphincter repair, and a prior lateral episiotomy is possibly a good prognostic factor. PMID- 11053948 TI - Biliary stricture due to hypertrophied liver rotation after right hepatic lobectomy. AB - BACKGROUND: After partial hepatectomy, liver regeneration occurs with the hepatocyte mass to its almost previous size. Biliary stricture due to hypertrophied liver rotation following partial hepatectomy is uncommon and its management still remains controversial. We encountered extrahepatic biliary stricture due to hypertrophied left lobe rotation in 2 patients who had right hepatic lobectomy for liver trauma. Herein, we present our experience with the modified Longmire procedure for the management of this complication. RESULTS: There were no complications during the early postoperative period in either of the patients. After the Longmire procedure, the first patient was followed for 5 months with some elevation in liver enzymes and the other was followed for 14 months with normal liver function tests. Both of the patients are completely symptom free postoperatively. CONCLUSION: The present experience suggests that the modified Longmire procedure is the most promising surgical approach to the management of biliary stricture due to hypertrophied left lobe rotation after right hepatic lobectomy. PMID- 11053949 TI - Association of transjugular intrahepatic portosystemic shunt with embolization in the treatment of bleeding duodenal varix refractory to sclerotherapy. AB - BACKGROUND: Bleeding from duodenal varices are often severe (mortality as high as 40%), and more difficult to sclerose than esophageal varices. We report a patient with a bleeding duodenal varix, refractory to sclerotherapy, successfully treated by the association of portosystemic shunt placement and varix embolization, via the same transjugular intrahepatic route. METHODS: A 40-year-old Black male underwent emergency TIPS and duodenal varix embolization after failure of endoscopic sclerotherapy. The portosystemic pressure gradient droped from 16 to 9 mm Hg following TIPS. At 5 months from TIPS, the patient is well, with a patent shunt at Doppler ultrasound. CONCLUSION: The present report of successful control of duodenal varix, actively bleeding and refractory to sclerotherapy, by means of combined TIPS and embolization, supports the role of TIPS and suggests that its association to embolization can be valuably considered in the difficult setting of portal hypertension with bleeding duodenal varices. PMID- 11053950 TI - Fatal intestinal perforation secondary to fragmentation of a Celestin tube. AB - BACKGROUND/AIMS: Oesophageal intubation remains one of the principal methods of palliation for an obstructing oesophageal carcinoma. We present a case which illustrates a rare but fatal complication of this procedure. METHODS: A 60-year old female with oesophageal cancer presented with total dysphagia 9 months following insertion of a Celestin tube for palliation. Oesophagoscopy revealed a bolus obstruction which was successfully cleared. Two days later she developed generalised peritonitis and subsequently died. RESULTS: A post-mortem examination demonstrated fragmentation and displacement of the distal part of the Celestin tube resulting in perforation of the small bowel. CONCLUSION: Celestin tube disintegration is a risk associated with long-term use, and routine replacement is indicated in patients with a prolonged survival to avoid this complication. PMID- 11053951 TI - Noninfectious gas accumulation in an infarcted spleen. AB - BACKGROUND: Intentional selective occlusion of the arterial blood supply to tumors of abdominal organs is a well established therapeutic procedure. Several reports described gas accumulation at the infarcted sites. These gas collections are usually nonsuppurative; however, the differential diagnosis should include abscess formation. CASE REPORT: We present a 59-year-old patient in whom the splenic artery was accidentally ligated during gastrectomy surgery, with resultant splenic infarction. Gas accumulation was diagnosed by sonography and CT studies. To the best of our knowledge this is the first report ever published in the English medical literature describing nonsuppurative gas formation within an abdominal organ, caused by accidental ligation of its main arterial supply during surgery. SUMMARY: Possible theories regarding this noninfectious gas accumulation are discussed and the differential diagnosis between abscess formation and noninfectious gas accumulation is emphasized. Establishing the correct diagnosis is of big clinical importance as the treatment of choice is completely different in each one of these entities although the imaging features, in ultrasound as well as in CT, are somewhat similar. PMID- 11053952 TI - Combined hepatic artery and segmental portal vein occlusion in antiphospholipid syndrome. AB - Antiphospholipid syndrome can have various clinical presentations, two of the most common being arterial and venous thrombosis. It is, however, unusual for them to occur in combination. We report here a case of combined hepatic artery and segmental portal venous occlusion in a 32-year-old patient who was shown to have a lupus anticoagulant. There have been no previous reports of thrombosis occurring simultaneously in the coeliac axis and the portal vein. Computerised tomography, Doppler ultrasound scanning and selective visceral angiography were used to demonstrate the anatomical lesions. The patient was treated medically with unfractionated heparin leading to a favourable clinical outcome. The diagnosis and management of this case is discussed with reference to the current literature on visceral thrombosis and antiphospholipid antibody syndrome. PMID- 11053953 TI - Gastrointestinal autonomic nerve sarcoma presenting as a giant intra-abdominal cyst. AB - Gastrointestinal autonomic nerve sarcomas are rare tumours. A 36-year-old man presented with abdominal pain, distension and constipation. Abdominal ultrasound and CT scanning demonstrated a giant cystic intra-abdominal mass. Laparotomy confirmed a large cystic mass arising from the ileum with multiple metastases. Immunohistochemical staining was positive for vimentin, neuronal-specific enolase and PGP9.5. This is the first reported case to present as a giant intra-abdominal cyst. Specialist histopathological and immunohistochemical analysis is essential to establish the diagnosis of this rare tumour. PMID- 11053954 TI - Surgical treatment of leiomyosarcoma of the distal duodenum. AB - Malignant tumors of the small intestine are rare. An uncommon finding of leiomyosarcoma located in the fourth part of the duodenum was diagnosed by gastrointestinal contrast studies, CT and angiography. Although malignant lesions of the small bowel are usually diagnosed late and thus are far advanced, curative resection was possible in our case. The location and histology of the tumor permitted a 'pancreas-preserving segmental duodenectomy'. The operative approach and exposure using the Cattell maneuver is described. It is emphasized that the more extensive pancreatoduodenectomy should be reserved for adenocarcinomas or lesions situated in the proximal part of the duodenum. Thirteen years following the operation, the patient is asymptomatic while CT and gastrointestinal contrast studies reveal no evidence of disease. PMID- 11053955 TI - Needle perforation of the appendix. AB - BACKGROUND/AIMS: Needles are among the most frequently swallowed foreign bodies. In most cases they are excreted per vias naturales, but in some cases needles can lead to perforation of the stomach or duodenum in adults. METHODS: We report a case of acute appendicitis after perforation of the appendix by a swallowed needle. An appendectomy was performed without any knowledge of the reason for perforation. RESULTS: Inspection of the resected appendix demonstrated a needle of 1.5 cm length inside the lumen of the appendix, which had caused a perforation of the distal end of the appendix. CONCLUSION: As seen in this case a longer lasting nondiagnosed perforation can lead to extensive local inflammation which could have been avoided by early surgical treatment which should also be considered if the patient has very few symptoms, as in this case. PMID- 11053956 TI - Pelvic actinomycosis presenting as ureteric and rectal stricture. AB - BACKGROUND/AIMS: Simultaneous ureteric and rectal stricture due to pelvic actinomycosis is very rare and only a few cases of either rectal or ureteric stricture have been reported. Our aim is to report a case of stricture of the rectum and the right ureter due to pelvic actinomycosis infection in a 63-year old man. METHODS: Explorative laparotomy and biopsies of the inflammatory pelvic mass were the only procedures that led to the definitive diagnosis of actinomycosis. Temporary diverting colostomy, drainage of the right ureter by a pigtail catheter and postoperative treatment with appropriate antibiotics were successful in eradicating the inflammatory process. CONCLUSIONS: Extensive pelvic masses involving pelvic viscera should be biopsied before undertaking any major surgery because of the possibility of pelvic actinomycosis. PMID- 11053957 TI - Pseudo-Meigs' syndrome caused by secondary ovarian tumors from gastrointestinal cancer. A case report and review of the literature. AB - BACKGROUND: Pseudo-Meigs' syndrome is a condition characterized by nonmalignant ascites and/or pleural effusion caused by pelvic tumors other than solid benign ovarian tumors. This syndrome has only rarely occurred in association with gastrointestinal cancers. METHOD: We treated a 53-year-old woman who developed this syndrome due to ovarian metastasis from colon cancer. Diagnostic work-up for abdominal distension disclosed a sigmoid colon cancer and bilateral ovarian masses. Ultrasonography demonstrated massive ascites and a right pleural effusion. Repeated cytologic examinations of both effusions revealed no malignant cells. Laparotomy disclosed no peritoneal dissemination. A radical sigmoidectomy and hysterectomy with bilateral salpingo-oophorectomy were performed. RESULTS: Histologic examination confirmed ovarian metastases from the colonic primary tumor. After resection, both effusions disappeared promptly, confirming a diagnosis of pseudo-Meigs' syndrome caused by sigmoid colon cancer. The patient remains alive with disease after 52 months. CONCLUSION: Among 6 reported occurrences with gastrointestinal tumors including our case, the primary site was the colon or rectum in 5 and the stomach in 1. Two cases were due to Krukenberg tumors. Three patients with documented outcomes were alive 108, 52, and 12 months after resection. Clinicians should note that gastrointestinal cancers, especially colorectal tumors, rarely may cause pseudo-Meigs' syndrome and resection may provide long-term palliation. PMID- 11053959 TI - Periodic arm movements in patients with the restless legs syndrome. AB - A high proportion of patients with restless legs syndrome (RLS) also complain of arm paresthesia but the presence of periodic arm movements (PAM) has never been documented in a sleep laboratory in these patients. We investigated the prevalence of PAM during nocturnal sleep and awakenings in 22 RLS patients. Fifteen patients had a PAM index >5 movements per hour during wakefulness and among them only 3 had a PAM index >5 during sleep. Twenty patients had a periodic leg movement (PLM) index >5 during wakefulness and 17 had a PLM index >5 during sleep. In 42.8% of cases, PAM showed temporal relationship with PLM during wakefulness. These results show that PAM is frequent in RLS and suggest that the basic neurological dysfunction responsible for RLS is probably not located exclusively at the level of the lumbar spinal cord but involves neuronal systems located at upper levels. PMID- 11053960 TI - Remote effects of chronic botulinum toxin treatment: electrophysiologic results Do not indicate subclinical remodelling of noninjected muscles. AB - This study investigates the remote effects of botulinum toxin injections by examining the motor unit architecture of noninjected distant muscles. In 21 dystonia patients treated with botulinum toxin (n = 11, mean cumulative dose = 815 mU; n = 10, mean cumulative dose = 7,207 mU) and 10 control individuals, a blinded single-fiber electromyography of the vastus lateralis muscle was performed. The main outcome measure was fiber density (FD), thus measuring the effect of different cumulative doses on remote reinnervation. FD was normal in all patients treated with botulinum toxin. FD did not differ between the three groups studied. No relationship was found between FD and cumulative dose. Therefore, in this specific patient population, muscles remote to the site of injection showed no FD change months after the injection. We conclude that there was no evidence of remote reinnervation and remodelling of motor units with cumulative chemodenervation. PMID- 11053961 TI - Long-term remission of idiopathic cervical dystonia after treatment with botulinum toxin. AB - Botulinum toxin type A (BTX-A) treatment for cervical dystonia is traditionally considered a purely symptomatic treatment. BTX-A blocks acetyl choline exocytosis for 3-6 months and most patients require reinjection after this period. We report on 6 patients (mean age 41.6 years, range 18-69) with idiopathic cervical dystonia who were treated with BTX-A injections and became asymptomatic for 2-4 years. Four patients showed remission after the first BTX-A treatment, 1 patient after the second set of injections and 1 after the third session. Amelioration of neck dystonia was observed within 1-4 weeks after the last BTX-A treatment and all 6 patients are symptom-free, off antidystonic medications for over 2 years. The possibility that BTX-A treatment may increase the chances of development of clinical remission in patients with idiopathic cervical dystonia is discussed. PMID- 11053962 TI - Effect of bilateral deep-brain stimulation on oral control of patients with parkinsonism. AB - We assessed the oral system of parkinsonian patients treated with bilateral chronic stimulation of the ventral intermediate nucleus of the thalamus (Vim), and of the subthalamic nucleus (STN) to compare the effects of these procedures on the articulatory organs. Force transducers were used to sample ramp-and-hold force contractions generated by the upper lip, lower lip and tongue at 1- and 2 newton target force levels, as well as maximal force. Fourteen healthy control subjects and 14 parkinsonian patients participated in this study. After an overnight fast the patients were evaluated under two conditions: during bilateral stimulation and 1 h after stopping stimulation. With Vim stimulation, dynamic and static control of the articulatory organs was worsened while with STN stimulation it was greatly improved. The target of stimulation differently influences the oral system and modulates neuronal structures involved in speech. PMID- 11053963 TI - Posttraumatic focal dystonia of the shoulder. AB - Posttraumatic movement disorders remain a controversial issue with focal dystonia being a prominent representative. Focal dystonia of the shoulder without concomittant cervical dystonia is a rare event. We describe 2 patients who, after minor trauma, developed focal dystonia of the shoulder with severe chronic pain. Response to local botulinum toxin A was favorable in 1 patient. PMID- 11053964 TI - 'Iatrogenic' Wernicke's encephalopathy in Japan. AB - 'Iatrogenic' Wernicke's encephalopathy has appeared to occur more frequently in Japan, probably induced by the change of our Japanese national health insurance policy in 1992. We report 4 nonalcoholic patients with such Wernicke's encephalopathy, which occurred during the early postoperative oral food intake period following intravenous nutrition without vitamin supplements. We analyzed the medical records of 4 patients, 3 men and 1 woman, aged between 55 and 71 years, who were admitted to our hospital between 1992 and 1995. Three patients underwent gastrointestinal surgery and 1 suffered chronic pyothorax. We diagnosed our patients as having Wernicke's encephalopathy based on typical neurological abnormalities, in addition to typical cranial magnetic resonance image findings, low serum vitamin B(1) levels, or both. Although all of the patients were treated with vitamin B(1) and showed some improvement, 1 patient developed Korsakoff syndrome, 2 made incomplete neurological recovery, and 1 died. We speculated that the body vitamin B(1) stores had been decreasing in our patients who did not receive any vitamin supplements during intravenous hyperalimentation or hydration. Subsequent administration of high calorie and high carbohydrate oral diets increased the demand for vitamin B(1), further depleting the vitamin stores, thereby causing 'iatrogenic' Wernicke's encephalopathy. The change of our national health insurance policy in 1992 discouraged the routine administration of vitamins, probably causing Wernicke's encephalopathy in our patients. PMID- 11053965 TI - Lymphocyte subset proportions in Guillain-Barre syndrome patients treated with plasmapheresis. AB - We examined the changes in the proportions of certain T cell subsets in 18 Guillain-Barre syndrome patients treated with plasmapheresis (PP). PP was performed within 8 days after the onset using a continuous-flow blood cell separator or immunoadsorption. The proportions of CD3+, CD4+ and CD4+CD45RA+ cells at the onset were decreased and that of CD19+ was increased compared with those of 25 normal controls. However, the proportions of CD4+CD45R-, CD4+HLADR+, CD4+CD25+ cells and the CD8+ subpopulations were similar in patients and controls. After PP, the altered subsets tended to normalize, particularly in the group that improved after PP. PMID- 11053966 TI - Spinocerebellar ataxia in Singapore: predictive features of a positive DNA test? AB - Significant differences in frequency of the different spinocerebellar ataxia (SCA) subtypes have been described to occur in different populations. A 'blunderbuss' diagnostic DNA testing approach would entail unnecessary healthcare cost. In this study, we determine the prevalence of SCA subtypes and predictive features of a positive DNA test in consecutive clinically diagnosed SCA cases in Singapore. Twenty-one consecutive patients from 14 families were evaluated over a 3-year period. Thirteen patients (61.9%) from 6 families had a positive DNA test. Eleven of these (all ethnic Chinese) had SCA 3 (abnormal CAG size ranged from 61 to 71), and 2 ethnic Malays had SCA 2 (abnormal CAG size of 39). Clinical features which were highly predictive of a positive DNA SCA test in our population included presence of a positive family history, chorea and dystonia, muscle and tongue fasciculations, gaze-evoked nystagmus, and hypertonia. Our study draws attention to the observation that knowledge of relatively specific features of the most common SCA subtype in a local population can greatly enhance the practical accuracy of the choice of which SCA DNA test to order. PMID- 11053967 TI - Alterations in trk A, trk B and trk C receptor immunoreactivities in parietal cortex and cerebellum in Alzheimer's disease. AB - The neurotrophins nerve growth factor, brain-derived neurotrophic factor and neurotrophin-3 bind to the tyrosine kinase (trk) receptors trk A, trk B and trk C, respectively, with high affinity. We investigated the expression of the trk receptors in the parietal cortex (PC) and cerebellum of patients with Alzheimer's disease (AD) and age-matched controls. Cortical layers II-VI displayed a distinct cellular immunoreactivity for trk A and C with an emphasis in the pyramidal neurons of layers III and V. Trk B immunoreactivity was primarily located in the deeper cortical layers with a predominance in layer V. There was a decrease in trk A and C immunoreactivity in the PC of AD cases, while trk B density appeared to be unchanged. In addition, cerebellar Purkinje cells revealed a distinct immunoreactivity for trk C both in control and AD cases, suggesting trk C may be important in the maintenance of these cells in the aged brain. PMID- 11053968 TI - Holocord syringomyelia as the dominant feature in Noonan's syndrome. PMID- 11053969 TI - Acute disseminated encephalomyelitis following Legionella pneumophila infection. PMID- 11053970 TI - Successful treatment with alpha-interferon of a patient with chronic measles infection of the brain and parkinsonism. PMID- 11053971 TI - Paroxysmal leg weakness and hearing loss in a patient with subarachnoid hemorrhage. PMID- 11053972 TI - Neurological involvement in Werner's syndrome: clinical and biopsy study of a familial case. PMID- 11053973 TI - A small 55-repeat MJD1 CAG allele in a patient with Machado-Joseph disease and abnormal eye movements. PMID- 11053974 TI - Genetic models of hypertension in experimental animals. AB - Genetic animal models are central to ongoing efforts to elucidate the pathophysiology and genetic basis of hypertension. The rat is the leading species in experimental hypertension. Several rat models of hypertension are available for research, including inbred strains, congenic lines, transgenic animals and recombinant inbred strains. Each of these models has been designed to express different phenotypes, including spontaneous hypertension, salt sensitivity, stress sensitivity and susceptibility to end-organ damage. All these models have been extremely useful in the search for the physiological mechanisms that underlie hypertension, but some of them have been specifically designed for detecting the hypertension genes. This latter task is extremely complex in spontaneous hypertension, but genetic animal models may simplify the task by enabling to focus on specific phenotypes. Despite intensive efforts over nearly 3 decades, the genetic basis of hypertension has not been unveiled so far in the rat or in other species. Recent dense mapping of the rat genome, the development of new strategies and technologies in molecular genetics including differential gene expression, expressed sequence tags and DNA biochips render hope that the formidable task of identification of new candidate genes in hypertension will move another major step forward. Once these genes are identified, their function and role in hypertension will have to be determined, utilizing functional genomic strategies and bioinformatics. Finally, the findings in genetic animal models of hypertension will have to be extrapolated to humans by homology and syntenic mapping strategies. PMID- 11053975 TI - Nephrotoxicity of angiotensin inhibition during the perinatal period. AB - As the only ex utero mechanism for the removal of nitrogenous waste, the mammalian kidney achieves some 50-fold increase in urine production during the perinatal period when the placental circulation becomes no longer available as a functional dialyzer. This urine is efficiently removed from the kidney by the renal pelvis, a smooth muscle structure unique to mammals, which develops during the perinatal period. We found that mutant mice completely devoid of angiotensinogen or its type 1 receptor, as well as wild-type neonates given an ACE inhibitor, fail to develop a renal pelvis or a ureteral peristaltic movement. These structural and functional defects in the urinary tract are followed by severe obstructive injury of the renal parenchyma. The ability of angiotensin to directly induce the pelvis is demonstrated in an organ culture system, in which treatment with angiotensin induces the characteristic smooth muscle layer in the wild type, but not in homozygous null mutants. Upregulation of both renal angiotensin content and type 1 receptor at the renal hilum are also demonstrated in the wild type during the transition from intra- to extra-uterine life. By inducing the timely development of the renal pelvis, angiotensin thus facilitates the removal from the renal parenchyma of the urine that promptly increases at birth, thereby effectively preventing a buildup of intrarenal pressure and a consequent development of dysmorphic kidney. PMID- 11053976 TI - Interactions between mitochondrial proteins and lipid peroxidation products in the maintenance of the glomerular filtration barrier in the in vitro perfused kidney. AB - BACKGROUND: The fourth complex of the mitochondrial respiratory chain, cytochrome c oxidase (CytC) consists of thirteen both mitochondrially and nuclearly encoded subunits, which are differently regulated in proteinuric kidneys. The effect of mitochondrial involvement on proteinuria is not known. METHODS: We set up an in vitro kidney perfusion model to study the direct effect of inhibitors of the mitochondrial respiratory chain, rotenone and antimycin A, on the glomerular filtration barrier by using immunohistochemistry and Northern blotting and quantitating the resulting proteinuria. RESULTS: Rapid onset of proteinuria and characteristic changes in CytC subunits were seen in the perfused kidneys. Urinary protein excretion increased significantly in the rotenone- and antimycin A-treated groups during perfusion. Downregulation of CytC subunits I and IV was similarly found in the groups treated with rotenone and antimycin A, while increases in the lipid peroxidation (LPO) products malondialdehyde and 4 hydroxynonenal which reflect mitochondrial damage, were observed. CONCLUSIONS: These data show rapid changes in mitochondrial proteins and induction of proteinuria in response to exposure to mitochondrial inhibitors. Together with the concomitant increase in local LPO products, these results suggest that continuous maintenance of a proper energy balance is important to maintain the glomerular filtration barrier. PMID- 11053977 TI - ACE inhibition preserves heparan sulfate proteoglycans in the glomerular basement membrane of rats with established adriamycin nephropathy. AB - The gradual onset of the antiproteinuric effects of ACE inhibition suggests that structural effects on the glomerular basement membrane (GBM) may be involved in their renoprotective action. To test this hypothesis, we studied the effects of lisinopril (5 mg/kg/24 h) on proteinuria, focal glomerulosclerosis (FGS) and glomerular heparan sulfate (HS) proteoglycan (HSPG) GBM staining in rats with established Adriamycin nephrosis. Treatment was started 6 weeks after disease induction. As expected, lisinopril reduced blood pressure, proteinuria and the FGS score. In control rats, Adriamycin nephrosis was associated with significantly impaired GBM staining for both HSPG core protein (assessed from BL 31 staining) and HS staining (assessed from JM-403 staining) 12 weeks after disease induction. In rats treated with lisinopril (5 mg/kg/24 h) GBM staining was significantly better preserved for HS as well as for HSPG core protein. These data suggest that structural effects on the GBM, improving glomerular permselectivity, may be involved in the renoprotective effects of ACE inhibition in proteinuria-induced renal damage. PMID- 11053978 TI - Oxidant stress activates AP-1 and heparin-binding epidermal growth factor-like growth factor transcription in renal epithelial cells. AB - Ischemia/reperfusion injury increases the expression of bioactive heparin-binding epidermal growth factor-like growth factor (HB-EGF) in the rat kidney, suggesting that oxidant stress or cell injury related to oxidant stress might affect HB-EGF expression in the injured renal parenchyma. We utilized a nontransformed rat renal epithelial cell line (NRK-52E cells) to investigate whether reactive oxygen species induced transcriptional activation of HB-EGF mRNA. Hypoxia/reoxygenation increased HB-EGF expression in NRK-52E cells, and at concentrations that induced sublethal cell injury, hydrogen peroxide (H(2)O(2)) increased HB-EGF mRNA expression 4.7-fold. The free radical scavengers, dimethylthiourea and N acetylcysteine inhibited HB-EGF mRNA induction. In contrast, another free radical scavenger, pyrrolidine thiocarbamate (PDTC), augmented H(2)O(2)-mediated HB-EGF expression. Since PDTC has been reported to augment AP-1-mediated transcriptional activation, we utilized an electrophoretic mobility shift assay to confirm that H(2)O(2) administration to NRK-52E cells did increase nuclear extract DNA-binding activity to a consensus AP-1 sequence. Using a CAT reporter assay coupled to the proximal 2,000 bp of the human HB-EGF 5'-untranslated region, we determined that H(2)O(2) administration increased CAT activity 5.5-fold. Truncation or deletion mutations of a putative AP-1-binding site reduced the H(2)O(2)-stimulated activity by >60%, and there was increased DNA binding of nuclear extracts from H(2)O(2)-treated cells to a 24-bp oligonucleotide containing this putative AP-1 site. Anti-fos and jun antibodies inhibited this binding, and there was no binding to an oligonucleotide in which the putative AP-1 site was mutated. The site of the residual activation was found to exist in the most proximal 5' untranslated region (-121 to +60), which contains two putative SP1 sites. Timing and localization of AP-1-binding activity from nuclear extracts from the post ischemic tissue correlated with HB-EGF mRNA expression. Therefore, in renal epithelial cells, oxidant stress increases HB-EGF expression, which appears to be mediated in part by an increase in AP-1 binding. This activation may play an important role in the induction of HB-EGF mRNA in response to tissue injury and may regulate early stages of recovery following ischemic damage. PMID- 11053979 TI - Renal and urinary glycosaminoglycans in an experimental model of chronic renal failure in rats. AB - The present paper reports the glomerular and renal individual glycosaminoglycan levels in an experimental model of chronic renal failure (CRF) that was induced in Wistar rats by five-sixths mass ablation. Glycemia, body weight, blood systolic pressure and urinary excretions of creatinine, albumin and glycosaminoglycans were measured for 12 weeks. At the end of the experiment, the weight and the glycosaminoglycan composition of the kidneys were determined. In control rats, heparan sulfate was the main glycosaminoglycan found both in whole kidney and isolated glomeruli, with trace amounts of dermatan sulfate. Isolated glomeruli presented higher heparan sulfate concentrations than whole kidney (expressed as mg/g dry weight). In CRF rats, albuminuria appeared from the 2 week on, and dermatan sulfate and chondroitin sulfate contents of the kidney increased, whereas heparan sulfate levels remained unaltered. Changes in urine glycosaminoglycans (heparan sulfate, chondroitin sulfate and dermatan sulfate) were not statistically significant. The increase in glomerular dermatan sulfate and chondroitin sulfate observed in this experimental model could be related to the mechanisms involved in the glomerulosclerosis and proteinuria that occur in CRF. PMID- 11053980 TI - Reduction in mean glomerular pore size coincides with the development of large shunt pores in patients with diabetic nephropathy. AB - The glomerular size selectivity was determined by neutral dextran clearance sieving technique and plasma cystatin C levels in two groups of patients with long-standing type I diabetes mellitus and different stages of albuminuria but normal glomerular filtration rate and in a group of healthy controls. The sieving characteristics of the glomerular filter were determined using a mathematical model of log normal pore size distribution. Patients with albuminuria above 200 microg/min exhibited a significant increase of cystatin c plasma concentrations and a significant reduction in mean glomerular filtration slit size. Only these patients exhibited large, unrestrictive pores in the glomerular filter (shunt). The plasma cystatin c levels correlated significantly with 26-angstrom neutral dextran plasma levels in microalbuminuric patients and in patients with albuminuria above 200 microg/min. We conclude that a reduction in average pore size of the glomerular filter does not occur earlier than the development of large shunt pores. The renal clearance of cystatin c in patients with proteinuric diabetic nephropathy but a normal glomerular filtration rate is reduced due to its molecular size. PMID- 11053981 TI - Low leptin mRNA level in adipose tissue and normoleptinemia in experimental chronic renal failure. AB - BACKGROUND: Anorexia and weight loss frequently accompany chronic renal failure (CRF). Although multiple metabolic changes occur during CRF, a bulk of evidence indicates that the decrease in caloric intake plays a major role in CRF-induced weight loss. Recently, it has been suggested that elevated plasma leptin concentrations could contribute to anorexia and to downregulation of leptin gene expression in CRF patients. However, in some CRF patients, plasma leptin concentrations have been found to be lower than one could expect. Thus we assumed that inhibition of leptin synthesis plays an important role in the regulation of plasma leptin concentrations in CRF patients. METHODS: To test this assumption, the leptin mRNA level in rat white adipose tissue from ad-libitum-fed control (sham operated), pair-fed control (sham operated) and rats with experimentally induced CRF has been measured by Northern blotting analysis. In addition, serum leptin concentration (by radioimmunoassay) was determined in all three groups of animals. RESULTS: The results of the present study indicate that in experimental CRF the leptin mRNA level is decreased by about 50% as compared to the sham operated animals (ad-libitum-fed and pair-fed controls). The mean serum leptin concentration in CRF rats was essentially similar to the leptin concentration in sham-operated ones. CONCLUSION: The data obtained suggest that in CRF animals the serum leptin concentration might be affected not only by the decrease in leptin removal in the kidney, but also by the decrease in leptin secretion from adipose tissue. Furthermore, the results of the study suggest that leptin may be only one of many factors involved in the pathogenesis of malnutrition associated with CRF. PMID- 11053982 TI - Intrarenal renin-angiotensin system contributes to tubular acidification adaptation following uninephrectomy. AB - BACKGROUND/AIMS: Reduction in renal mass by uninephrectomy induces a functional compensation in the remnant kidney. The activity of the angiotensin-converting enzyme (ACE) as well as renin mRNA in the proximal convoluted tubule (PCT) of uninephrectomized (UNx) rats increases. The aim of this work was to determine whether the increased activity of the local renin-angiotensin system (RAS) participates in the adaptation of renal function after uninephrectomy. METHOD: We utilized normal two-kidney (2K) and 3-week UNx rats to study the activity of the ACE in vesicles obtained from luminal membranes of proximal tubular cells and the acidification kinectics in PCTs using micropuncture techniques. RESULTS: The converting enzyme activity was significantly larger in UNx (5.87+/-0.69 nmol x min(-1) x mg protein(-1)) than in 2K rats (2.43+/-0.13 nmol x min(-1) x mg protein(-1); p<0.05). The acidification rate constant (kappa) in PCT of 2K rats was 0.18+/-0.02 s(-1) and in UNx rats 0.30+/-0.04 s(-1) (p<0.001). In UNx rats, microperfusion with 10(-5) M ramipril or 10(-5) M losartan decreased kappa to 0.19+/-0.02 and 0.18+/-0.02 s(-1), respectively, but had no effect on 2K rats. Luminal steady-state pH (pH(infinity)) was the same in 2K and UNx rats, and was not modified by addition of 10(-5) M ramipril or 10(-5) M losartan in both groups. The proximal H(+) flux (J(H(+))), calculated from pH(infinity) and kappa, was 1.12 nmol x cm(-2) x s(-1) in 2K rats and, 1.77 nmol. cm(-2). s(-1) in UNx rats (p<0.001). In 2K rats, this value was not changed by 10(-5) M ramipril or 10(-5) M losartan, but in UNx rats J(H(+)) decreased 25 and 30% with ramipril or losartan, respectively (p<0.001). CONCLUSIONS: These data suggest that the increase in the local RAS activity could be an adaptive change that contributes to maintain the homeostasis of body fluids after uninephrectomy. PMID- 11053983 TI - Temozolomide in patients with high grade gliomas. AB - The incidence of central nervous system neoplasias ranges from 3.8 to 5.1 cases per 100,000 inhabitants. First-line treatment of brain tumors consists of surgery associated with radiotherapy, and followed or not by chemotherapy. When used as an adjuvant therapy after surgery and radiotherapy, chemotherapy prolongs the time to progression and the median survival time. At relapse, chemotherapy is the common therapeutic approach. In recent years, new drugs for the treatment of brain tumors have been tested, and some of them, such as temozolomide, are very promising. PMID- 11053984 TI - Oxaliplatin-induced fever and release of IL-6. AB - BACKGROUND: Oxaliplatin is a novel cytotoxic agent with documented activity in colorectal cancer. Side effects are generally moderate, and include peripheral neuropathy along with mild bone marrow suppression and gastrointestinal side effects. To our knowledge, induction of febrile episodes by this agents has not been described in the literature. CASE REPORT: We present the case of a 74-year old male patient admitted to our institution for palliative treatment of metastatic colorectal carcinoma. Due to progression during treatment with 5 fluorouracil and leucovorin, chemotherapy consisting of oxaliplatin 85 mg/m(2) on days 1 + 15 plus mitomycin C 8 mg/m(2) on day 1 repeated every 28 days was initiated. The first cycle of this combination was tolerated without side effects, but the patient experienced fever up to 39 degrees C starting 2 h after oxaliplatin administration on day 15 of the second cycle, which persisted for 3 days. Fever again recurred at the same interval following administration of oxaliplatin on day 1 of the next cycle. Blood samples taken at regular intervals disclosed an increase in IL-6 serum levels parallel to the body temperature curve, with the peak corresponding to the highest temperature, while C-reactive protein values remained unchanged. In spite of intensive premedication with steroids, antipyretics and clarithromycin, fever promptly recurred during the third cycle of treatment. CONCLUSION: Our data suggest a clear- cut correlation between fever, the release of IL-6 and oxaliplatin administration. Whether IL-6 release is directly triggered by the application of oxaliplatin or is a bystander phenomenon, however, remains unclear at the moment. PMID- 11053985 TI - A unique case of eccrine porocarcinoma with pulmonary lymphangitis and pericardial involvement: biological characterization and clinical aggressiveness. AB - A unique case of eccrine porocarcinoma with pulmonary lymphangitis and pericardial involvement is reported. The clinical course was aggressive, leading to the death of the patient a few months after diagnosis. Certain pathologial markers of clinical aggressiveness were retrospectively investigated: p53 and Ki 67 expression were determined by means of immunohistochemistry. Angiogenesis was assessed by determination of intratumor microvessel density at the vascular 'hot spot' with the anti-CD34 monoclonal antibody and quantitative analysis using computerized image analyzer. Both primary tumor and metastatic lymph node presented immunostaining for p53 and Ki-67, with a higher degree of vascularization in the secondary lesions compared to the primary tumor. Our findings suggest a correlation between tumor vascularization and clinicopathological parameters of aggressiveness in malignant eccrine porocarcinoma. Taking into account the disappointing results of current treatments for metastatic eccrine porocarcinoma, the assay of microvessel density may be helpful in selecting the patients of high risk for recurrence or death who may benefit of anti-angiogenic therapies. PMID- 11053986 TI - Clinical investigation on pulmonary metastasis of head and neck carcinomas. AB - OBJECTIVE: To establish a treatment strategy for pulmonary metastases, we clinically investigated the characteristics of distant metastases from head and neck carcinomas. METHODS: In 636 head and neck carcinomas, the pathophysiology of distant metastases was investigated by charts, roentgenographies, computed tomographies and scintigraphies. RESULTS: Of the squamous cell carcinomas, oropharyngeal tumors were most highly metastatic, followed by lower gingiva, floor of the mouth, maxillary sinus, and tongue. In distant metastases, 30 (4.7%), 5 (0.8%), and 7 (1.1%) metastasized to the lungs only, lungs and other organs, and organs excluding the lungs, respectively. In pulmonary metastases, the right, left and both lungs were involved in 18, 5, and 8 patients, respectively, although details were not obtained for 4 patients. Pulmonary metastases consisted of 1, 2, and 3 or more tumors in 18, 4, and 6 patients, respectively. Diffuse cancer cell infiltration was observed in 3 patients. Of the 42 patients with distant metastases, 12 patients died of progressive pulmonary metastases, and 5 of these patients manifested only 1 pulmonary lesion throughout life. However, the metastatic pulmonary tumors were controlled surgically or conservatively in 3 patients. CONCLUSION: These results indicate that distant metastases from head and neck carcinomas involve the lungs most frequently and that chemoimmunotherapy and surgical removal of the metastatic tumors are recommended when indicated. PMID- 11053987 TI - Oral doxifluridine in advanced hepatocellular carcinoma: A phase II study. AB - Hepatocellular carcinoma (HCC) remains one of the most common neoplasms in the world. Doxifluridine is an oral fluoropyrimidine derivative activated to 5 fluorouracil by uridine phosphorylase which is more expressed in malignant cells. Therefore, we conducted a phase II study to evaluate the activity of oral doxifluridine in patients with advanced hepatocellular carcinoma. Twenty-five advanced hepatocellular carcinoma patients entered the study; doxifluridine was given orally at the initial daily total dose of 2,250 mg for 4 consecutive days every week. All patients are evaluable for toxicity: these included mainly grade 1-2 (WHO) diarrhea, stomatitis, nausea and vomiting; 4 patients (16%) experienced grade 3-4 diarrhea. Twenty-four patients are evaluable for response and 1 complete and 3 partial responses have been observed (response rat 17%, 95% confidence interval: 5-37). Oral doxifluridine at the dose and schedule we used, although having only modest activity in advanced HCC, may represent an alternative to other frequently used chemotherapeutic agents, because of its favorable toxicity profile and its simple route of administration. PMID- 11053988 TI - Analysis of genomic imprinting of insulin-like growth factor 2 in colorectal cancer. AB - Genomic imprinting of insulin-like growth factor 2 (IGF2) has been shown to play an important role in the development of Wilms' tumor and adult cancers including lung and esophageal cancer. Although IGF2 has been reported to be overexpressed in colorectal cancer, the status of this gene has not been fully elucidated. To clarify genomic imprinting of IGF2 in colorectal cancer, we examined loss of imprinting (LOI) and loss of heterozygosity (LOH) in surgically resected tissues by utilizing Apa1 polymorphism. Of 46 patients with colorectal cancer, 2 exhibited (2/15, 13%) LOH among 15 patients who were heterozygous for IGF2 DNA in nontumorous tissues. Four (4/12, 33%) patients showed LOI of IGF2 gene among informative patients. In these patients, the reverse transcription polymerase chain reaction and immunohistochemistry revealed that IGF2 mRNA is overexpressed in tumorous tissues, compared to nontumorous tissues. Of 15 patients in whom IGF2 was immunohistochemically more highly expressed in tumorous than in normal tissues, there were 2 with LOH and 4 with LOI. The results of the present study suggest that LOI of IGF2 plays an important role in the carcinogenesis of colorectal cancer. PMID- 11053989 TI - Fine needle aspiration may shed breast cells into peripheral blood as determined by RT-PCR. AB - OBJECTIVE: A diagnostic test applying reverse-transcriptase chain reaction (RT PCR) assay targeted against cytokeratin 19 (CK19), cytokeratin 20 (CK20) and the beta-subunit of human chorionic gonadotropin (beta-hCG) mRNAs was used to evaluate the impact of fine needle aspiration (FNA) on breast cell shedding into peripheral blood. METHODS: The sensitivity of this assay was based on the different degree of admix of MCF-7 breast cancer cell line with HL-60 leukemic cell line. For blood samples of 24 cases with benign breast diseases and 20 cases with malignant ones, 5 ml of peripheral blood was drawn before and within 10 min after puncture. Total RNA was extracted from peripheral blood mononuclear (PBMN) cells; beta-actin was used to assess the quality of cDNA. RT-PCR products were run in ethidium bromide gel and observed under ultraviolet. RT-PCR products for beta-hCG were digested with Sty I endonuclease to confirm the specificity. RESULTS: The sensitivity of RT-PCR assay was 1 MCF-7 cell in 10(5) HL-60 cells for CK19 and CK20, and 1 in 10(6) for beta-hCG. For 24 benign cases, none of the pre- FNA samples was positive for CK20 and beta-hCG, and 3 cases (12.5%) were positive for CK19. As for 20 malignant cases, 1 pre-FNA sample was positive for all three markers and 2 other samples were positive for CK19. After aspiration, 3/21 benign cases and 1/17 malignant case with pre-FNA negative signals became positive for CK19, while 3/19 malignant cases with pre-FNA negative signals were converted to a positive result for CK20 and beta-hCG. Of 6 pre-FNA positive cases, all cases remained positive for the respective marker. CONCLUSION: FNA to breast tumor may cause hematogenous dissemination of breast cells. PMID- 11053990 TI - Reduced expression of manganese superoxide dismutase mRNA may correlate with invasiveness in esophageal carcinoma. AB - Little is known about the expression and antioxidant function of manganese superoxide dismutase (Mn-SOD) in esophageal squamous cell carcinoma. To determine the significance of Mn-SOD in esophageal squamous cell carcinomas, Mn-SOD mRNA expression was examined in 45 esophageal squamous cell carcinomas and the corresponding normal mucosal tissues by reverse transcription-polymerase chain reaction. The tumor/normal (T/N) ratio of 45 patients with esophageal carcinoma was calculated, and the data were clinicopathologically analyzed. The T/N ratio of Mn-SOD mRNA expression was less than 0.5 in 11 (32.4%) of 34 esophageal carcinoma cases without any preoperative treatments, while none of 11 cases who underwent preoperative chemotherapy showed a T/N ratio of <0.5 (p < 0.05). There was an inverse correlation between the Mn-SOD expression level and the degree of venous invasion (p < 0.05) as well as lymphatic invasion (p < 0.05). Furthermore, poorly differentiated squamous cell carcinoma showed significantly lower Mn-SOD mRNA expression levels than well differentiated carcinoma (p < 0.05). Our results suggest that Mn-SOD mRNA was frequently reduced in esophageal carcinoma when compared to the normal mucosa and the reduced expression levels of Mn-SOD mRNA may lead to an accumulation of superoxide radicals in conjunction with the increased invasiveness of esophageal carcinoma. PMID- 11053991 TI - Growth characteristics of rectal carcinoid tumors. AB - PURPOSE: Tissue growth depends on both cell proliferation and cell death. This study was designed to examine the growth characteristics of rectal carcinoid tumors. METHODS: Fifty rectal carcinoid tumors were studied clinicopathologically and experimentally. Expression of Ki-67, TGF-alpha, p53, and bcl-2 was examined immunohistochemically, and apoptotic cells were identified by the in situ DNA nick end labeling method. EGF receptor expression was examined by a colorimetric in situ mRNA hybridization technique. RESULTS: The median Ki-67 labeling index (LI) in all lesions was 0.62 +/- 0.59%. Ki-67 LI was significantly (p < 0.01) higher in lesions larger than 5 mm than in lesions smaller than 5 mm. TGF-alpha was expressed more frequently (p < 0.01) in lesions larger than 5 mm (100%) than in lesions smaller than 5 mm (65.2%). Ki-67 LI was significantly (p < 0. 05) higher in lesions with TGF-alpha expression than in lesions without TGF-alpha expression. The in situ hybridization revealed EGF receptor expression in all 46 lesions with intact mRNA (100%), and coexpression of TGF-alpha and EGF receptor was found in 39 of the 46 (84.8%) lesions. The median apoptotic index (AI) in all lesions was 0.15 +/- 0.12%. AI has increased with tumor size and was significantly (p < 0.05) higher in lesions with a higher Ki-67 LI than in lesions with a lower Ki-67 LI. p53 protein was detected in only 1 patient who had liver metastases, and the gene mutation was confirmed by polymerase chain reaction and single-strand conformation polymorphism analysis. bcl-2 expression was absent in all lesions. CONCLUSIONS: The Ki-67 LI indicated a low cellular proliferative activity in rectal carcinoid tumors. AI was very low, and was significantly correlated with proliferative rate. Inhibition of apoptosis by mutated p53 or bcl 2 may not have occurred in most of these tumors. TGF-alpha/EGF receptor autocrine mechanisms may play a possible role in tumor growth, and the cellular proliferative activity may increase as tumors grow larger. PMID- 11053992 TI - Role of fibrinogen covalently associated with cell membrane in blood-borne lung tumor colony formation of murine mammary carcinoma cells. AB - The role of fibrinogen covalently associated with cell membrane in blood-borne lung tumor colony formation of murine mammary carcinoma cells in mice was studied. When mice were treated with prednisolone, their plasma fibrinogen levels profoundly increased. Hyperfibrinogenemia, induced by prednisolone treatment or plasma fibrinogen infusion of syngeneic mice, accelerated the coagulation time and significantly increased the number of lung tumor colonies of SCK tumor cells. Hypofibrinogenemia, induced by rabbit antisyngenic mouse fibrinogen immunoglobulin G or heparin infusion, markedly delayed coagulation time and prominently reduced the numbers of blood-borne lung tumor colonies of the tumor cells. SCK mammary carcinoma cells form a coating of fibrinogen on their surfaces in a medium containing fibrinogen. This coating is cross-linked in a manner characteristic of catalysis by tumor cell membrane-bound transglutaminase K. The fibrinogen coating on the surface of these tumor cells functions to protect against autologous lymphokine-activated killer cells. These results provide information on the impact of fibrin stability on blood-borne lung tumor colony formation of SCK mammary carcinoma cells. PMID- 11053993 TI - Suppression of invasion and MMP-9 expression in NIH 3T3 and v-H-Ras 3T3 fibroblasts by lovastatin through inhibition of ras isoprenylation. AB - Lovastatin, a hydroxymethylglutaryl coenzyme A reductase inhibitor, was found to block the synthesis of cholesterol and to affect posttranslational modification or isoprenylation, which is essential for membrane localization and biological activity of several proteins including Ras in the signal transduction pathway. Ras activates a multitude of downstream activities with roles in cellular processing, including invasion and metastasis. We investigated the anti-invasive activity of lovastatin in NIH 3T3 and v-H-Ras-transformed NIH 3T3 (v-H-Ras 3T3) cells. Lovastatin suppressed cell invasion in vitro in a dose-dependent manner. By zymographic assay, a decrease in matrix metalloproteinase-9 (MMP-9) activity but not matrix metalloproteinase-2 (MMP-2) activity by lovastatin was detected. RT-PCR demonstrated a reduction in gene expression of MMP-9 after treatment with lovastastin. To confirm the lovastatin-induced down-regulation of MMP-9 expression, we transfected an MMP-9/luciferase reporter vector, under MMP-9 promoter control, into both NIH 3T3 and v-H-Ras 3T3. A reduction in luciferase activity was observed with lovastatin treatment. In addition, lovastatin also reduced AP-1 and NFkappaB binding activities. These anti-invasive features were attenuated by the presence of mevalonate. These results suggest that down regulation of MMP-9 contributes to the anti-invasive activity of lovastatin. Furthermore, we added exogenous mevalonate, which enhances the potency of cell invasion, and Ras farnesyltransferase inhibitor (manumycin A), which inhibits the potency of cell invasion. In accordance, Western blot analysis showed that lovastatin decreased membrane localization of Ras proteins. These data indicate that the anti-invasion activity of lovastatin happens through a decrease in Ras isoprenylation and functions. PMID- 11053994 TI - Enhanced selective gene expression by adenovirus vector using Cre/loxP regulation system for human carcinoembryonic antigen-producing carcinoma. AB - Selective gene targeting using the carcinoembryonic antigen (CEA) promoter is useful in gene therapy for gastrointestinal cancer. However, the expression of the CEA promoter is not sufficient. In this study, we tried to enhance CEA promoter activity using the Cre/loxP system. The double infection of CEA producing cells such as MKN45 and LoVo with AxCEANCre and AxCALNLZ at a total multiplicity of infection (MOI) of 50 achieved 7-fold higher expression level of beta-galactosidase activity than single infection of those cells with AxCEALacZ at 50 MOI. On the other hand, the double infection of CEA-nonproducing cells such as MKN1 and HeLa cells showed a very low expression of beta-galactosidase activity. In the subcutaneous tumor models, the administration of AxCEANCre and AxCALNLZ into the CEA-producing tumor showed stronger expression of the LacZ gene in tumor tissue than that of AxCEALacZ. In the experiment using orthotopic models of CEA-producing gastric cancer, intraperitoneal double administration of AxCEANCre and AxCALNLZ caused evident LacZ gene expression in transplanted gastric tumors, but no LacZ gene expression in the normal stomach or liver. It was confirmed that enhanced tissue-specific gene transduction under control of CEA promoter using the Cre/loxP system was useful not only in vitro, but also in vivo, especially in orthotopic models. PMID- 11053995 TI - Diabetic retinopathy: surrogate outcomes for drug development for diabetic retinopathy. AB - Diabetic retinopathy remains the most frequent cause of new cases of blindness among adults aged 20-74 years. A number of large clinical trials have validated treatment methods now considered standard. However, the disease continues to progress in approximately 50% of the eyes treated by photocoagulation. Other forms of therapy targeted at the earliest stages of retinal disease are needed. The difficulties in defining and accepting surrogate outcomes appropriate to evaluate the earlier stages of retinopathy are discussed. Special attention is given to trial design and the most likely possibilities for surrogate outcomes: mean difference on the Early Treatment Diabetic Retinopathy Study retinopathy scale of at least 2 steps per eye, reduction in macular thickening and reduction in fluorescein leakage. PMID- 11053996 TI - Intubation of the lacrimal pathways under endoscopic control. AB - Described herein is a new modification of the lacrimal pathway intubation in children with efferent lacrimal pathway obstruction. For removal of the intubation set probe without olive tips from the inferior nasal meatus, a special titanium loop is used. An endoscopic technique allows us a basically more considerate performance of this procedure and to speed it up. The risk of iatrogenic damage to the mucous membrane of the inferior meatus is, thus, reduced too. From April 1994 to June 1998, lacrimal pathway intubation was performed in 235 children 283 times. Infraction of the inferior concha was performed 27 times. Insertion of a probe submucously into the inferior nasal meatus was found 35 times and the probe was released by focused incision of the mucous membrane. PMID- 11053997 TI - Arcuate relaxing incisions with a 5.00-mm optical zone for the correction of high postcataract astigmatism. AB - The purpose of this study was to investigate the efficacy and safety of arcuate cuts with a 5-mm optical zone to correct high postoperative astigmatism after extracapsular cataract extraction. We performed 5-mm optical zone arcuate cuts on 23 eyes of 23 patients with high postoperative astigmatism. Ophthalmic examination included uncorrected visual acuity (UCVA), best spectacle corrected visual acuity (BSCVA) and the amount of the refractive and keratometric cylinder before and 9 months after operation. Surgically induced refractive change was calculated in all cases. A significant reduction in astigmatism was achieved in all cases with minimal axis deviation. No case developed clinically significant irregular astigmatism. The mean magnitude of astigmatism of the surgically induced refractive change calculated from standard keratometry and refractive data was 3.73+/-0.72 and 3.70+/-0.77 dptr, respectively. The mean axis deviation calculated from the keratometric and refractive data was 1.18+/-2.33 and 1.32+/ 3.62 degrees, respectively. At the last examination, 78.2% of the eyes had UCVA of 20/40 or better. No eye lost more than two lines of vision, 6 eyes lost one line, 2 eyes gained one and 1 eye gained two lines of BSCVA. The above data show that the 5-mm optical zone arcuate astigmatic keratotomy is an effective and safe method of correcting high postoperative astigmatism. PMID- 11053998 TI - High hurdle of clinical trials to demonstrate efficacy of anticataractogenic drugs. AB - Recently, the rapid progression of cataract surgical technique has led cataract patients in industrialized countries to ignore the possibilities of drug therapy. Globally, however, it will be impossible in the near future to treat cataract by surgery alone, mainly due to medicoeconomic reasons. Preventative measures must be sought. As one of the these measures, the development of anticataractogenic drugs has reemerged as a focus in the lens research field. Although clinical trials of newly developed drugs are absolutely necessary before they enter the market, they have been considered to be a rather easy task. However, in order to gain accurate and reproducible data from trials, the trial program must be carefully prepared. The numbers of participants to the trial, the selection criteria of the subjects, the objective judgment of cataractous changes, follow up period, a high technical level for cataract documentation and image analysis are proposed. Although there still remain some difficulties concerning the methods for objective judgment, a scientifically acceptable examination must be conducted. PMID- 11053999 TI - Ocular findings in aplastic anemia. AB - OBJECTIVE: To analyze the ocular findings in aplastic anemia. DESIGN: Eighteen patients with aplastic anemia were examined. RESULTS: Ocular findings included cotton wool spots (38%), nerve fiber layer or preretinal hemorrhages (67%), vitreous hemorrhages (13%), a picture resembling central retinal vein occlusion (13%) and optic disk edema (6%). Preretinal hemorrhages were the presenting sign of aplastic anemia in 2 patients (13%). CONCLUSIONS: A blood profile is needed in patients with unexplained retinal hemorrhages. Patients with aplastic anemia need to avoid ocular massage and Valsalva maneuvers to decrease ocular morbidity. PMID- 11054000 TI - Visual acuity disturbances in chronic renal failure. AB - Patients affected by chronic renal failure often complain of blurred vision when submitted to hemodialysis. Refraction, visual acuity and lens transparency have been evaluated in 36 eyes of 18 patients who underwent hemodialysis, before and after the treatment. Student's t test did not prove any statistically significant difference between the considered parameters. However, a change in refraction was noted in 64% of the eyes, always in hyperopic mean. Corrective glasses had to be changed to ensure the same visual acuity as before the hemodialytic treatment. Particular care must be taken in lens prescription in those patients who could undergo dialysis for chronic renal failure. PMID- 11054001 TI - Idiopathic juxtafoveolar retinal telangiectasis: clinical pattern in 19 cases. AB - BACKGROUND: Idiopathic juxtafoveolar telangiectasis (IJT) is a rare retinal vasculopathy of adulthood that may cause central loss of visual acuity. The purpose of this paper is to evaluate the type and distribution of fundus changes in patients with this disorder. METHODS: Medical records of 19 patients with IJT using the Gass & Blodi classification were studied. Eight patients in group 1A (42%) and 11 in group 2A (58%) were analyzed. By projecting color photographs we divided the macula into four quadrants and studied the fundus changes in each one. RESULTS: All patients in group 1A presented unilateral disease with easily seen telangiectasis, lipid deposits and cystic macular edema. The mean age of this group was 55.8 years. Five (63%) patients were male and 3 (37%) were female. All patients in group 2A, 2 males (18%) and 9 females (82%), had bilateral disease evidenced by fluorescein angiography. The mean age of this group was 54.5 years. Fundoscopic findings included: intraretinal pigment plaques, grayish coloring of the macular retina, superficial crystalline deposits, right-angle venules, subretinal and intraretinal neovascularization. CONCLUSIONS: All patients in this survey were adults. In group 1A, men were affected 2 times more often than women, and the main affected macular quadrant was the inferior temporal, followed by the inferior nasal. In group 2A, women were affected 4 times more often than men and all eyes had inferior and superior temporal macular quadrant involvement. PMID- 11054002 TI - The Reykjavik Eye Study--prevalence of lens opacification with reference to identical Japanese studies. AB - PURPOSE: A population-based cataract epidemiological study, the Reykjavik Eye Study, was conducted to determine the prevalence and characteristics of lens opacities in Iceland. The results were compared with those from previous surveys in Japan to determine the influence of race and environmental factors. METHODS: 1,635 randomly selected residents of Reykjavik, Iceland, were enrolled, and among them, 1,045 responded and took part in the study. The subjects received ophthalmological examination of both the anterior and the posterior segments. Changes in the crystalline lens were examined and photographed under maximal pupillary dilation. Classification of the opacity types and grading of the extent of lens opacificiation were done using a standardized scheme, and the data were analyzed with Student's test, the chi(2) test and the Mantel-Haenszel test. RESULTS: The prevalence of lens opacities in subjects in their 50s, 60s, 70s and in those 80 years and older was 42.7, 61.0, 85.3 and 100%, respectively. Grade II and III lens opacities were found in 2. 2% of subjects in their 50s and in 10.5, 35.9 and 62.3% in their 60s, 70s, and over 80 years, respectively. Cortical opacities predominated in all age groups followed by nuclear opacities; subcapsular opacities, on the other hand, were quite rare. A unique type of opacity, i.e., granular opacity, seldom seen among the Japanese, was found in 6.6% of the Icelandic subjects. CONCLUSIONS: Our results show that cortical opacity is the major type of opacity in Icelandic subjects, although its prevalence was lower than that in the Japanese subjects. Interestingly, the first appearance of lens opacities of grades II and III in Icelanders was delayed by 10 years compared to the Japanese suggesting fewer or less severe risk factors for cataract formation in Iceland. PMID- 11054003 TI - Bilateral superior cervical ganglionectomy increases choroidal blood flow in the rabbit. AB - In this study, we evaluate the effect of bilateral superior cervical ganglionectomy on albino rabbit choroidal blood flow (CBF) in changes of perfusion pressure (PP). Twenty albino rabbits of either sex weighing between 2.0 and 3.0 kg were randomly divided into two groups. The experimental group (group S) included 10 rabbits (20 eyes) that received bilateral superior cervical ganglionectomy one week prior to the study. The other 10 rabbits (20 eyes) served as a control (group N): each received the same procedure as group S except that the superior cervical ganglion was preserved. By means of a laser Doppler flowmeter (Perimed PF4001), blood cell flux (PF), velocity (V), and concentration of moving blood cells (CMBC) were recorded simultaneously while intraocular pressure was increased linearly by a syringe pump. Blood pressure and intraocular pressure were continuously monitored. The laser beam focused on the posterior pole away from major retinal vessels. When PP decreased from 75 to 0 mm Hg, in group N, PF, V and CMBC decreased from 100% to 6.94+/-0.91%, 8.41+/-0.87%, and 19.38+/-1.11%; in group S, it was 17.75+/-2.58%, 16.78+/-1.48%, and 34.58+/ 4.42%, respectively. Group S poses higher PF, V, and CMBC than group N while PP decreased gradually. These results indicate that the superior cervical ganglion plays a role in CBF regulation. Bilateral sympathectomy led to a higher PF in the group S rabbits, indicating increased CBF. PF, V, and CMBC remained constant until PP <55 mm Hg demonstrated the existence of CBF autoregulation within a limited perfusion range in both groups. This autoregulation did not change after sympathectomy. PMID- 11054004 TI - Remission of superior oblique myokymia after microvascular decompression. AB - Superior oblique myokymia (SOM) is an ocular motility disorder characterized by oscillopsia, vertical or torsional diplopia, sometimes combined with pressure sensation. Although the pathophysiological basis is unclear, isolated case reports have documented its association with intracranial pathological processes. We present a case of SOM associated with a vascular compression of the fourth nerve at the root exit zone. Following microneurosurgical decompression, SOM completely resolved and paralysis of the fourth nerve occurred. This was less disturbing. PMID- 11054005 TI - Effective treatment with topical cyclosporin A of a patient with Cogan syndrome. AB - The purpose of this report is to describe the effective treatment of severe anterior segment inflammation due to Cogan syndrome through the use of topical administration of cyclosporin A. A 47-year-old female patient had been experiencing headaches and difficulties with her vision. Subsequent examination revealed the sudden onset of bilateral conjunctival injection and swelling of bilateral auricles. Despite the multiple treatment (systemic and topical corticosteroid and antibiotic therapy), necrotizing scleritis had appeared bilaterally and the scleral wall was thinning. Topical administration of 1% cyclosporin A was applied to both eyes 4 times a day. After 2 months of this therapy, the epithelial tissue covered the necrotizing tissue and her symptom of ocular pain was relieved and her corrected visual acuity was improved. This is the first case exhibiting that topical cyclosporin A is an effective treatment for severe anterior segment inflammation associated with Cogan syndrome. PMID- 11054006 TI - Clinical diagnosis of chronic canaliculitis by 20-MHz ultrasound. AB - The practical value of high-resolution ultrasound (transducer frequency of 20 MHz) in the study of the lacrimal canaliculi has been proven. It can also be used in the clinical diagnosis of chronic canaliculitis. If the classic symptoms are absent, the clinical diagnosis is often inaccurate, and treatment is insufficient. Representative images of normal cases and of chronic canaliculitis illustrate the potential of high-resolution ultrasound. In our patient, 20-MHz scanner images revealed pathological findings which were invisible during slitlamp examination. Ultrasonic images of chronic canaliculitis showed ectasia of the canaliculus and sulfur grains. High-resolution ultrasonic examination of the lacrimal drainage system demonstrated that the 20-MHz scanner used was able to show concrements (sulfur grains), measuring 1-2 mm in diameter. Such more reflective structures (like sulfur grains) are a pathognomonic sign of chronic canaliculitis. Our report confirms the efficiency of 20-MHz sonography in the diagnosis of canaliculitis without any side effects. PMID- 11054007 TI - Juvenile xanthogranuloma of the orbit in an adult. AB - We present a case of juvenile xanthogranuloma (JXG) with unilateral involvement of the orbit and eyelid and proptosis, histologically confirmed in a 32-year-old man with a 1-year history of a pansinusitis and dacryoadenitis with rhinitis. Nine months later an infiltration of the anterior upper part of the right orbit and right eyelid appeared. Computed tomography scan and magnetic resonance imaging studies confirmed the presence of pansinusitis and infiltration. The patient underwent a blepharoplasty and excision of the infiltrated tissues of the orbit, eyelid, and levator muscle. Hematoxylin-eosin and immunohistochemical studies revealed features consistent with a diagnosis of JXG (Touton giant cells). JXG, a non-Langerhans'-type benign proliferation, is a rare condition in adulthood. PMID- 11054008 TI - Pattern dystrophy of the retinal pigment epithelium in Crohn's disease. A case report. AB - The authors describe a case of bilateral pattern dystrophy of the retinal pigment epithelium in a man with Crohn's disease. The patient was examined every 6 months over a follow-up of 30 months. The right eye presented a macroreticular dystrophy while in the left eye a butterfly pattern dystrophy was diagnosed. During the follow-up period the retinal lesion changed; in the right eye the lesion increased in size, while in the left eye the morphology of the lesion passed from the butterfly to Sjogren's type. This report adds a new ocular manifestation of Crohn's disease, emphasizing the importance of the ophthalmological follow-up in the recognition of posterior segment complications associated with this inflammatory bowel disease. PMID- 11054009 TI - Controversies in the treatment of N(0) neck in laryngeal cancer: neck dissection, no surgery or sentinel lymph node biopsy? PMID- 11054010 TI - Stereoscopic display of a three-dimensional image of the larynx using high-speed helical scanning. AB - To evaluate the usefulness of stereoscopic images of larynges using helical CT in stereo mode, a retrospective review of the characteristics of stereoscopic viewing of larynges was made. The subjects were 3 patients with laryngeal cancer, 1 patient with laryngeal leiomyosarcoma and 1 patient with an advanced tongue carcinoma whose formalin-fixed larynx was extirpated. The larynges were scanned by high-speed helical CT using 1- to 2-mm slices. The reproduction of stereographic images was performed by the manipulation and rotation of three dimensional structures around the y-axis on the computer display. The three dimensional images of the complex structures, such as the arytenoid cartilage, aryepiglottic fold and pyriform sinus, were better observed by binocular images (stereograms) than by monocular images. Stereoscopic views of the larynx are useful in producing three-dimensional images of the unseen inner surface of the human body. PMID- 11054011 TI - Malignant salivary gland tumors in Quito, Ecuador. AB - OBJECTIVES: Malignant salivary gland tumors (MSGT) are uncommon. Age-standardized incidence rates are 0.5 and 0.3 per 100,000 in Quito, Ecuador; and 1.0 and 0.7 per 100,000 in the USA (SEER Program), for males and females, respectively. The goal of this study was to review a 16-year experience of a major general hospital in the treatment of these lesions. PATIENTS AND METHODS: From 1982 to 1998, 308 salivary gland tumors were surgically treated at the Hospital 'Carlos Andrade Marin' of the Ecuadorian Institute of Social Security in Quito, Ecuador, an Andean city of approximately 2 million inhabitants. Malignant lesions were found in 58 cases (19%): 37 out of 194 parotid gland tumors (19%), 7 out of 86 submandibular tumors (8%) and 14 out of 28 minor salivary gland tumors (50%). Adenoid cystic carcinoma and mucoepidermoid carcinoma were the most common histologic types. Twenty-two cases were classified as stage I, 13 as stage II, 1 as stage III and 20 as stage IV (UICC TNM staging classification). Thirty-one (53%) patients were treated by surgery alone; postoperative radiation therapy was additionally given to 22 (38%), and surgery, radiotherapy and chemotherapy were applied in 5 cases (9%). RESULTS: Local (LR) and/or regional recurrences were detected in 13 patients (22%). Twelve patients (21%) developed distant metastasis (DM; 2 in more than one site): 7 in the lungs, 2 in the brain, 2 in the bone and 1 each in the liver, subcutaneous tissue and pleura. Thirty-five patients are alive, 33 disease free. Twenty-three patients are deceased: 6 with LR, 7 with DM, 3 with both LR and DM, 1 with locoregional recurrence and DM, 2 with a second neoplasm, 3 with intercurrent disease and 1 from unknown causes. Five- and 10 year overall survival rates were 75 and 68%, respectively. There were no significant differences in mortality according to the site of the primary tumor or histologic type, but stage and involved surgical margins were important prognostic factors (p = 0.006 and 0.003). CONCLUSIONS: The surgical or multimodality treatment of MSGT has provided a good locoregional control (78%) and 68% 10-year survival in a series of patients treated at the oncology department of a general hospital in Quito, Ecuador. Stage and involved surgical margins were significant prognostic factors. PMID- 11054012 TI - Nasal challenge activates the mucociliary transport system on not only the ipsilateral but also the contralateral side of the nose in patients with perennial allergic rhinitis. AB - It is controversial whether the mucociliary transport system in the nasal mucosa of persons with allergic rhinitis is upregulated after an allergen challenge in vivo. The aim of this study was to assess the effect of a specific allergen challenge on the mucociliary transport system in allergic rhinitis. We used a challenge of house dust mites or a control substance on one side of the nose of 97 volunteers (54 with allergic rhinitis for house dust mites and 43 without allergies). The mucociliary transport system of the ipsilateral or contralateral side of the nose challenged was evaluated using the saccharin clearance time on the right side of the nose. The saccharin clearance time was significantly shortened after an allergen challenge not only on the ipsilateral (right) side of the nose (from 15.0 +/- 11.6 to 10. 9 +/- 9.2 min, mean +/- SD, p = 0.041) but also on the contralateral (left) side of the nose (from 14.7 +/- 10.1 to 8.1 +/- 7.9 min, p = 0.013) in the allergic group. A reduction of the saccharin clearance time was also seen in the allergic group after a control challenge [ipsilateral (right) challenge: from 13.0 +/- 10.0 to 11.6 +/- 10.6 min, contralateral (left) challenge: from 13.5 +/- 9.0 to 10.2 +/- 10.1 min]. There were no reductions of the saccharin clearance time after an allergen or control challenge in the nonallergic groups. These results suggest that a unilateral nasal challenge accelerates the mucociliary transport system bilaterally in the nose of subjects with a hypersensitivity such as allergic rhinitis. PMID- 11054013 TI - Efficacy of inhalation form of furosemide to prevent postsurgical relapses of rhinosinusal polyposis. AB - The aim of the work is to demonstrate the efficacy of furosemide to prevent relapses of rhinosinusal polyps after surgical treatment. Two groups of people with rhinosinusal polyposis were enrolled: the study group consisted of 64 patients and the control group of 40 subjects. After surgical treatment, the study group started the therapy with topical furosemide; the control group had no treatment administered after the operation. Six years after the operation only 4 cases of relapse were noticed in the study group (10%), while there were 12 relapses, 4 slight (6.4%) and 8 severe (20%), in the control group. In conclusion, furosemide could represent a valid therapeutic aid in the prevention of nasosinusal polyps. PMID- 11054014 TI - The relationship between traumatic tympanic membrane perforations and pneumatization of the mastoid. AB - We evaluated the possible relationship between tympanic membrane perforations resulting from blast trauma or slap and pneumatization of the mastoid cells. A total of 25 male patients with tympanic membrane perforations resulting from blast injury (n = 7), slap (n = 17), and football hit (n = 1) and 20 healthy male volunteers without any ear problem had temporal bone computed tomographic scans in the axial plane, parallel to the infraorbitomeatal line, with 2 mm slice thickness and 2-mm intervals using bone algorithm with a ProSpeed Spiral tomography machine. The area of air cells in each slice was measured using trace and area measurement functions of the tomography machine, and by multiplying the resulting area by slice thickness, the volume of each slice was calculated. For each ear, the total of volumes of air cells was calculated by adding the volumes of each slice containing air cells. The calculated volumes of mastoid cells were evaluated by comparing microscopic findings. Both patient and control groups consisted of males, and their ages ranged from 17 to 32 (mean 24.5) years. Microscopic examinations revealed that perforations were frequently located in the lower quadrants and that most of them were less than 3 mm. There were no pars flaccida and marginal perforations. Ossicular chain destruction was noted neither in temporal bone tomographic nor during intraoperative examinations. The mean (+/ SD) volumes of right and left ear mastoid air cells in patient and control groups were 6.92 +/- 2.45 vs. 7.00 +/- 2.59 cm(3) and 9.04 +/- 4.55 vs. 8.95 +/- 4.53 cm(3), respectively, and the differences were not statistically significant. It was found that the level of mastoid pneumatization has no statistically significant effect on tympanic membrane pathologies due to blast or other injuries. PMID- 11054015 TI - Middle ear imaging using virtual endoscopy and its application in patients with ossicular chain anomaly. AB - Virtual endoscopy (VE) is a recently developed technique to provide a realistic surface rendering of various organs, which can be applied to the use of three dimensional (3D) studies of several lesions. However, its advantages in otological disease have not been well investigated. In this study, we evaluated the application of VE in patients with ossicular chain anomalies. Virtual middle ear endoscopy was a time-saving method, however, we needed the appropriate technical procedures of algorithm and reconstruction spacing to generate accurate 3D images of ossicles. We obtained virtual surgical views of middle ear structures and related anomalies, and confirmed by intraoperative findings that these images were mostly compatible with the actual lesions of ossicles. VE allowed an identification of the anatomy of the ossicles and adjacent structures simultaneously. The elements of the stapedial crura were clearly visualized with VE images in 93.3% of normal ears. Pathological ossicular chain findings such as malleus or incus fixation, dislocation and disruption, except footplate fixation were investigated successfully. One possible procedure, using alterable CT value in the obtained VE images on the monitor, is proposed for further detection of fine lesions of the ossicles. These observations suggest that virtual middle ear simulations accurately represent major intraoperative findings. This technique may have an important role in preoperative planning, surgical training, and/or postoperative evaluation in otology. PMID- 11054016 TI - Speech understanding with the CIS and the n-of-m strategy in the MED-EL COMBI 40+ system. AB - Speech tests have been performed on 6 subjects for comparing the standard 12 channel continuous interleaved sampling (CIS) strategy (CIS12), the 7-channel CIS strategy (CIS7) and the 7-of-12 strategy in the MED-EL COMBI 40+ system. An ABAB experimental design was used whereby each strategy was reversed and replicated. Speech tests were performed in quiet (vowels, consonants, monosyllables, sentences) and noise (sentences). Results showed that for vowels, CIS12 is significantly superior to CIS7, for consonants and sentences CIS12, CIS7 and 7-of 12 performed equally well, and that for monosyllables 7-of-12 is significantly superior to both CIS12 and CIS7. In addition, 7-of-12 is superior to CIS7 by almost the same amount as CIS12, but in this case the difference is not significant. Further, all strategies have been found to be equally robust in noise with respect to sentence understanding. The differences between CIS12 and 7 of-12 on the one hand and CIS7 on the other hand may be attributed to decreased spectral resolution of the latter. The fact that - in contrast to what has been reported for the SPEAK strategy - 7-of-12 is equally robust in noise as the CIS strategies is explained by the use of higher stimulation rates, wider frequency bands and a higher percentage of channels stimulated in each cycle. PMID- 11054017 TI - Apoptosis in auditory brainstem neurons after a severe noise trauma of the organ of Corti: intracochlear GDNF treatment reduces the number of apoptotic cells. AB - We have studied the morphological and cellular changes in the cochlear nucleus (CN) after cochlear nerve degeneration and whether these changes can be prevented by rescuing the primary cochlear neurons from degeneration with local glial cell line derived neurotrophic factor (GDNF) treatment. Degeneration of spiral ganglion neurons was seen to lead to a reduction of the volume of the anteroventral cochlear nucleus (AVCN); the size of the cell nuclei in the AVCN also was reduced. No differences were observed in cell density. After intrascalar GDNF treatment the volume of the AVCN was significantly larger when compared to the untreated side, and the size of the cell nuclei in the AVCN was significantly larger on the treated side. After degeneration of spiral ganglion neurons, an increased number of apoptotic cell nuclei were seen in ipsilateral CN and superior olivary complex. This increase was significantly smaller after intrascalar GDNF treatment. Degeneration of primary cochlear neurons seems to lead to an increase in the number of CN neurons undergoing apoptotic cell death. This can be prevented partially by rescuing primary cochlear neurons from degeneration with local GDNF treatment. PMID- 11054018 TI - Antrolithiasis in the frontal sinus. AB - A very rare case of sinusitis with antrolithiasis in the left frontal sinus of a 63-year-old male patient is reported. Various conservative treatments had no effect on the decrease of his left frontal pain and of postnasal drip. Neither bacteria nor fungus were detected in the discharge. Computed tomographic scanning revealed several high-dense spots in an isodense shadow in the left frontal sinus. At first, endoscopic sinus surgery (ESS) was employed and a stony mass was detected in the nasofrontal duct and sinus. Patency of the nasofrontal duct was insured and a sticky paste and small masses were removed as well as possible. However, the flow of discharge from the frontal duct continued after surgery. We performed a second operation with extranasal approach and additional stones in the sinus were successfully removed. Most cases of antrolithiasis are caused by a foreign body or caseous sinusitis with fungus. The maxillary sinus is the most common site of disease in antrolithiasis. It is unknown why the present case of antrolithiasis was in the frontal sinus. In such cases of antrolithiasis or cases having pastelike contents in the frontal sinuses, we conclude that ESS may be an unsuccessful treatment and a classical surgical approach may be required. PMID- 11054019 TI - Benign and malignant oncocytoma of the salivary glands with an immunohistochemical evaluation of Ki-67. AB - We present two cases of benign oncocytoma derived from the parotid gland and minor salivary gland and one case of malignant oncocytoma from the parotid gland. The proliferative activity of the tumor cells was evaluated immunohistochemically for Ki-67. The average frequency of Ki-67-positive cells was 3.3% in the benign oncocytomas and 6.5% in the malignant oncocytoma. The higher frequency of Ki-67 positive cells in the malignant oncocytoma might reflect active cell proliferation. Ki-67 immunostaining may be useful in distinguishing a benign oncocytoma from a malignant oncocytoma. PMID- 11054020 TI - The versatility of cytokeratins as tumor markers. AB - Cytokeratins are particularly versatile tools in tumor biology both experimentally and clinically. Previously regarded as a rigid intracellular skeleton, the cytokeratins are now known to provide a more dynamic structural integrity within epithelial cells. This more flexible nature of cytokeratin function occurs through rapid shifts in assembly/disassembly states, geared by phosphorylations/dephosphorylations in the 'head' and 'tail' regions of the monomeric filaments. When released from proliferating or dying tumor cells, they provide a useful marker for epithelial malignancies, as evidenced by the number of available immunochemical assays for cytokeratins. Another significant area of interest that highlights their utility is the deposition of cytokeratins in the necrotic areas of tumors, providing an effective target for radioimmunotherapy and radioimmunolocalization. Moreover, the appearence of cytokeratin neodeterminants seems to be a very early event during apoptosis. Indeed, the involvement of specific cytokeratin degradation patterns resulting from the activity of caspases during apoptosis highlights yet another developing field of research devoted to these highly pleomorphic and complex structures. PMID- 11054021 TI - Immunohistochemical profiles of 30 monoclonal antibodies against cytokeratins 8, 18 and 19. Second report of the TD5 workshop. AB - In the first report of the TD5 workshop (TD5-1), the epitope specificities of 30 different monoclonal antibodies against cytokeratins 8, 18 and 19 were determined. This second report presents the immunohistochemical profiles of these antibodies using human appendix and normal skin for evaluation. Each antibody was tested by one or two different laboratories recruited from the Dutch Working Group on Immunohistochemistry and Cytochemistry. Eight different laboratories participated. The histological specimens were pretreated by the participants in three different ways for immunohistochemistry: microwave antigen retrieval in citrate buffer, enzymatic digestion to restore epitope exposure, no specific treatment (untreated paraffin-embedded samples), and tested blindly without knowledge of cytokeratin or epitope specificity of the antibodies at three different concentrations of 50, 10 and 1 microg/ml. Most of the tested antibodies (29/30) were useful in at least one pretreatment method, with microwave antigen retrieval being the most sensitive approach. For some antibodies, very high backgrounds were observed. Furthermore, it can be concluded that 11 MAbs performed well using all three staining protocols, including untreated paraffin embedded sections. Interestingly, all the antibodies with documented selected specificity towards cytokeratin 8 (i.e. 178, 191, 199, 202 and 206) are reactive with an immunodominant region corresponding to amino acids 340-365 on cytokeratin 8, which evidently is well-suited as target for immunohistochemical interactions. Similarly, three antibodies with the same capacity to react with untreated samples had specificity against cytokeratin 19 (i.e. 179, 197 and 204) in the corresponding region in this filament, i.e. amino acids 311-335, or the KS 19.1 epitope. None of the six antibodies against the other major cytokeratin 19 epitope (BM 19.21) were found useful for immunohistochemistry on untreated samples. The overall conclusions from the present investigation are that all cytokeratin-8-specific antibodies with defined epitope specificities were very useful. Only one of the major two epitopes on cytokeratin 19 seems to be available for efficient immunohistochemistry. Cytokeratin 18 exposes some epitopes outside the immunodominant region reactive with the antibodies 190, 203 and 205 which can be used for untreated samples. The implications of these findings are of significance both for diagnostic histopathology and for the biology of tumor marker epitope expression in tissues. PMID- 11054022 TI - Increased expression of protease M in ovarian tumors. AB - Proteases are known to play important roles in tumor invasion and metastasis. Protease M, which was originally identified by Anisowicz and colleagues in 1996, is a new member of the serine protease family. We also identified the protease M transcript in a differential PCR screen of ovarian tumors and have investigated its expression in 44 ovarian tumors (12 low malignant potential tumors, 32 carcinomas) and 10 normal ovaries using quantitative PCR. The PCR product was labeled with (32)P and a phosphoimager was used to determine the relative expression of the protease M gene compared to internal control beta-tubulin. mRNA expression levels of protease M were significantly elevated in 9 of 12 low malignant potential tumors and 30 of 32 carcinomas. Northern blot hybridization showed that the 1.7-kb protease M transcript was abundant in carcinoma but not detected in normal ovary. Immunohistochemical staining of normal ovary and ovarian tumor tissue sections with antibodies generated to protease M derived peptides corroborated the semi-quantitative PCR and Northern analysis data. Our results suggest that protease M is frequently overexpressed in ovarian tumors and may therefore contribute to the invasive nature or growth capacity of ovarian carcinomas. PMID- 11054023 TI - High levels of cytokeratin 19 fragments but no evidence of cytokeratins 1, 2, 10/11, 14 or filaggrin in the serum of squamous cell lung carcinoma patients. AB - The CYFRA 21-1 assay detects circulating fragments of cytokeratin 19, which is a sensitive marker for the diagnosis of lung cancers, particularly squamous cell carcinomas and adenocarcinomas. Epidermis-type proteins, such as cytokeratins 1, 2, 10/11 and 14 or filaggrin, are also expressed in squamous cell carcinomas. These could also be pertinent tumor markers, ideally as sensitive as CYFRA 21-1 and more specific for squamous cell lung cancer. To verify this hypothesis, using monoclonal antibodies produced in our laboratory, we developed immunoassays specific for these proteins. After optimization, the immunoassays were evaluated in sera from 91 controls and 138 patients with squamous cell lung cancer and compared to conventional tumor markers (CEA, SCC Ag and CYFRA 21-1). Less than 14% of the sera were above the lower limit of detection of the cytokeratin- and filaggrin-specific immunoassays. Moreover, part of these positive sera were induced by the presence of interfering heterophilic antibodies in sera. Thus, in patients with squamous cell lung cancer, we confirmed the high diagnostic sensitivity of CYFRA 21-1 (55.6%) but were unable to detect significant levels of epidermis-type cytokeratins or filaggrin. PMID- 11054024 TI - Comparison of Cyfra 21-1 and SCC assays in head and neck tumours. AB - Patients with head and neck tumours (HNT) have a high risk of early locoregional relapse that is difficult to diagnose. This study evaluated the usefulness of the serum Cyfra 21-1 assay compared to squamous cell carcinoma antigen (SCC) assay for monitoring such patients. Three hundred and twelve HNT patients, including 204 newly diagnosed patients, were followed up for a median of 446 days with serial serum assays for SCC and Cyfra 21-1. Untreated patients showed SCC and Cyfra 21-1 serum levels correlated with each other: concentration was correlated to clinical stage, tumour size (as T1 + T2 vs. T3 + T4) and nodal status. Cyfra 21-1, but not SCC, was related to the presence of metastases and the primary tumour site, with a univariate prognostic value for disease-free survival (p = 0.015). Cox's regression analysis showed that only Cyfra 21-1 was associated with a risk of relapse (p = 0.027). The random coefficient growth curve model applied to serial SCC and Cyfra 21-1 measurements of 111 patients showed that only Cyfra 21-1 exhibited a significant difference between patients with and without relapses. We found Cyfra 21-1 to be more closely related to initial clinical data and disease evolution than SCC, and therefore propose the use of Cyfra 21-1 for monitoring head and neck cancers. PMID- 11054025 TI - Selection of human single chain Fv antibody fragments binding and inhibiting Helicobacter pylori urease. AB - Single chain Fv antibody fragments (scFv) binding to purified Helicobacter pylori urease were selected from a nonimmune human antibody repertoire displayed on filamentous phage. After three rounds of screening on solid phase urease, 44 clones were found to bind the enzyme and four distinct scFv were identified by sequencing their heavy and light chain variable region genes (V(H) and V(L)). Two of the selected human scFv (scFv B4 and scFv D9) inhibited the activity of H. pylori urease with inhibitory constants (K(i)) of 7 and 2 microM, respectively. Their affinity (K(d)) for H. pylori urease as determined by surface plasmon resonance ranged from 17 to 42 nM. Both scFv were able to bind to urease present on the surface of living H. pylori organisms as demonstrated by flow cytometry analysis. The binding sites of scFv B4 and D9 were mapped by the use of two random hexapeptide libraries (X6 and CX6C) displayed on filamentous bacteriophage. The selected peptide sequences were shown to inhibit scFv binding to H. pylori urease and thus could be used in a vaccination strategy as epitopes mimicking (mimotopes) the region of urease recognized by these human scFv antibody fragments. PMID- 11054026 TI - Isoform expression of CD44 adhesion molecules, Bcl-2, p53 and Ki-67 proteins in lung cancer. AB - CD44, belongs to the cell adhesion molecule family and is expressed on cell surfaces in several isoforms which are generated by alternative splicing of messenger RNA. These splice variants have been shown in several cancer cell types and are thought to be involved in tumor progression. The aim of the current study was to evaluate the expression of selected CD44 variants on lung cancer cells of various histology and to compare these with other markers of tumor spread. Surgical samples of primary lung carcinoma of various histology were subjected to alkaline phosphatase-anti-alkaline phosphatase complex immunohistochemistry using a panel of monoclonal antibodies: anti-CD44 v5, v6, v7/8, v10, anti-Ki-67, anti Bcl-2 and anti-p53. Positive cells were scored in a semiquantitative way. The patients were subdivided into groups with and without metastases, as found during surgery. All CD44 variants tested could be demonstrated on lung cancer cells, but the incidence of particular isoforms varied, depending on lung cancer histology. In general, CD44 expression was highest in squamous cell tumors and lowest in anaplastic small cell carcinomas. Squamous cell cancers had high expression of v5 and v6 variants, while in anaplastic large cell and small cell carcinomas v10 was abundant. When Ki-67, Bcl-2 and p53 protein expression was compared to the incidence of CD44 variants, coincidence was found for v10 only. Most of the cases positive for v10 were also Ki-67 positive (p = 0.0146). In 12 cases with metastases, tumor cells had high v6 and Ki-67 expression, but these data were not significant compared to cases without metastases. Overall, these data suggest that v5 and v6 variants are of significance in squamous cell lung carcinoma, presumably in the promotion of metastasis, while in anaplastic small cell or large cell cancers only v10 expression seems to correlate with proteins associated with tumor growth and progression. PMID- 11054027 TI - Comparison of two prognostic markers for malignant melanoma: MIA and S100 beta. AB - It has recently been shown that the serum level of melanoma-inhibitory protein (MIA) provides useful information for the therapy and follow-up of patients with malignant melanoma. Previously, S100 beta has been described as a useful tumor marker for malignant melanoma. In this study, we compare the significance of the two markers in follow-up, therapy outcome and prognosis by measuring MIA and S100 beta serum levels in 50 melanoma patients. Serum levels were measured in patients with malignant melanomas of stages I-IV with at least 3 time points of measurement. Serial MIA and S100 beta measurements were obtained from 32 patients with stage IV disease in parallel to chemotherapy and from 18 patients with a history of stage I and stage II disease during follow-up. The response to chemotherapy in stage IV disease and relapse of melanoma during follow-up correlated with changes in MIA and S100 beta serum levels. In comparison, MIA revealed slightly higher specificity and sensitivity. In conclusion, both markers are useful for detection of progression from localized to metastatic disease during follow-up and for monitoring therapy of advanced melanomas. PMID- 11054028 TI - Assessment of voiding outcome, sexual function and quality of life two years following KTP/YAG hybrid laser prostatectomy. AB - OBJECTIVE: To assess the long-term outcome of patients undergoing KTP/YAG hybrid laser treatment for bladder outlet obstruction due to benign prostatic enlargement, in terms of symptomatic relief, complications, sexual function, patient satisfaction and acceptance of procedure. PATIENTS AND METHODS: The study recruited 148 patients prospectively. The hybrid laser treatment involved performing an initial bladder neck incision using KTP laser at 34 W followed by 4/6 point coagulation using NdYAG laser at 60 W. Patients were followed up till 2 years and assessed using uroflowmetry, International Prostate Symptom Score (IPSS), Patient Satisfaction Score (PSS), BPH impact index (BII) and the Danish Prostate Symptom Score (DAN-PSS) sexual function questionnaire. RESULTS: 137 patients were followed up for 2 years. There was a significant improvement in the maximum flow rate, IPSS and Quality of Life Scale (QLS). The mean BII (2.9) and the mean PSS (1.9) were low suggesting overall satisfaction with the procedure. The complications included urethral stricture (0.73%), bladder neck obstruction (2.15%) and retreatment (3.6%). 79.4% had a significant decrease in the ejaculate and 32.8% had a significant change in the strength of erections. On comparing the two groups (sexual function affected vs. not affected), the age, BII, IPSS and PSS were significantly higher (p < 0.05) in the group of patients that were affected. CONCLUSIONS: Following KTP/YAG hybrid laser prostatectomy the outcome for voiding is good and durable for up to 2 years. The patient satisfaction level following the procedure is high and the procedure well accepted. However, significant interference with sexual function occurs, which appears to be occurring in tandem with a poor voiding outcome. PMID- 11054029 TI - General state of health and psychological well-being in patients after surgery for urological malignant neoplasms. AB - OBJECTIVE: To evaluate the general state of health and the psychological well being in a group of 155 patients after surgery for urological malignant neoplasms. MATERIALS AND METHODS: Surgery was performed in 55 patients for renal cell carcinoma, in 54 for invasive bladder carcinoma, in 30 for adenocarcinoma of the prostate, and in 16 for squamous penile carcinoma. All patients were invited to self-compile the General Health Questionnaire (GHQ) - 12 items according to Goldberg and the Hospital Anxiety and Depression Scale. Results were compared with those in a group of patients who underwent retropubic prostatectomy for benign prostatic hyperplasia. RESULTS AND CONCLUSION: The general state of health was significantly more impaired in neoplastic patients than in the control group. Levels of anxiety were significantly higher but depression levels were similar in both groups. As far as the type of tumor is concerned, patients who underwent radical cystectomy for bladder carcinoma and those treated with partial penectomy for squamous penile carcinoma showed a significant impairment of the general state of health compared with controls. Higher levels of anxiety were observed in patients who underwent ileal conduit after radical cystectomy, in those treated with radical prostatectomy for prostate cancer and in those who underwent partial penectomy. Significantly higher levels of depression than in the control group were observed only in patients with ileal conduit. PMID- 11054030 TI - Combination therapy of imipramine with oxybutynin in children with enuresis nocturna. AB - PURPOSE: The treatment approach for enuresis is controversial due to the lack of consensus as to the exact causes of nocturnal enuresis. Despite various treatment modalities, pharmacotherapy still appears to be the common choice. The aim of this prospective study was the evaluation of the efficacy of combination therapy (imipramine and oxybutynin) in patients with enuresis nocturna. PATIENTS AND METHODS: This prospective study was done with 77 monosymptomatic nocturnal enuretics between July 1996 and December 1998. RESULTS: Even though there is no statistically significant difference between combination therapy (imipramine plus oxybutynin) and monotherapy, clinical data showed that combination therapy is more effective. CONCLUSIONS: We conclude that combination of imipramine with oxybutynin is a safe and new choice in the treatment of nocturnal enuresis. PMID- 11054031 TI - Expression of E-cadherin and catenins on testis tumor. AB - E-cadherin (ECD) is a homophilic Ca(2+)-dependent adhesion molecule associated with cell-to-cell interactions and normal growth. Recent reports have suggested that decrease or loss of ECD facilitates tumor progression and/or metastasis. ECD functions in a complex called an adherens junction, which includes several other proteins including alpha- and beta-catenin. In the present study, fresh-frozen sections from 32 testis cancers, 16 seminomas and 16 non-seminomatous germ cell tumors (NSGCT), were examined immunohistochemically. E-cadherin was not expressed on normal germ cells, but expressed on 3 (18.8%) of 16 seminomas and 10 (62.5%) of 16 NSGCTs, mainly on the epithelial component of teratoma cells. alpha-Catenin was detected on 0 (0%) of 13 seminomas and 4 (25%) of 16 NSGCTs. beta-Catenin was detected on 10 (71.4%) of 14 seminomas and 13 (81.2%) of 16 NSGCTs. ECD was detected significantly more frequently on NSGCTs than on seminomas. Immunoblot analysis confirmed the expression of ECD and beta-catenin in NSGCTs. Expression of ECD and catenins may reflect the degree of differentiation and provide some information on the character of the tumor. PMID- 11054032 TI - Effects of aging on mitochondrial enzyme activity of rat urinary bladder. AB - OBJECTIVES: Our previous study showed that aged rat bladders became fatigued faster than young bladders following repeated contraction induced by electrostimulation. One factor might be a lower energy-producing capability secondary to a decreased mitochondrial enzyme activity of the aged bladder. This study examined this possibility. MATERIALS AND METHODS: Mitochondria from 3- (n = 11) and 24-month-old (n = 10) Sprague-Dawley rats were isolated. Activities of the following enzymes were assayed: two key enzymes in the citric acid cycle, citrate synthase and malate dehydrogenase, and three enzymes in the respiratory chain reaction, NADH-cytochrome c reductase, succinate-cytochrome c reductase and cytochrome c oxidase. The concentration of phosphocreatine and ATP in the aged rat bladders and a separate group of young bladders (n = 12) was determined using high-performance liquid chromatography. RESULTS: (1) The aged bladders have a significantly lower level of phosphocreatine and ATP content than those of young bladders. (2) The activities of all five enzymes assayed were significantly lower in the aged bladders than in young bladders, especially for citrate synthase, which had only 46.8% of the activity of young bladders. CONCLUSIONS: Aging reduces the mitochondrial enzyme activity of the rat bladder resulting in a lower energy-production capability, which might explain some of the voiding dysfunctions found in the elderly. PMID- 11054033 TI - Effect of endothelin receptor antagonist (TAK-044) on autotransplanted perfused kidneys in dogs. AB - BACKGROUND: Endothelin-1 (ET-1) is involved in some diseases, including renal disease. Recently, the role of ET-1 in postrenal transplantation has been demonstrated in experimental and clinical studies. A new endothelin receptor antagonist, TAK-044, blocks both, ETA and ETB receptors, and was useful in treating acute renal failure in rats. In this study, we evaluated the effect of TAK-044 on autotransplanted kidneys with 18 h of perfusion. MATERIALS AND METHODS: TAK-044 was injected subcutaneously at 15 mg/kg/day for 2 weeks in one group of dogs, and blood analysis and renal function, were evaluated. A control group was given saline in the same manner as that used for the TAK-044 group. Histopathological examination and immunohistochemistry for ET-1 were performed in the two groups. RESULTS: In the control group, 5 of the 7 dogs died of renal failure within 2 weeks after autotransplantation of the kidney. In the TAK-044 group, 5 of the 7 dogs survived and 2 died of renal failure within the same period. Although the histological changes in the tubules in both groups were severe due to the 18 h of perfusion, TAK-044 ameliorated these changes. Immunohistochemical staining for ET-1 was seen in tubules in the control group. CONCLUSION: These findings suggest that TAK-044 effectively reduces damage in autotransplanted perfused kidneys in dogs, and may be useful in limiting damage to the kidney by acute tubular necrosis after renal transplantation in humans. PMID- 11054034 TI - Italian urological manpower: employment prospects and future scenarios. A panorama of employment opportunities for young urologists. AB - INTRODUCTION: Italy delayed limiting access to the Faculty of Medicine and the reform of schools of specialization was not accompanied by programming the number of scholarships, so employment expectations are often disappointing. The aim of this study is to analyze the employment prospects of specialists in urology through the development of possible scenarios for the 5-year period 2000-2004. MATERIALS AND METHODS: We recorded data received from Italian Schools of Specialization in Urology on specialists in the 5-year period 1994-1998. We also tried to obtain a picture of the national distribution of urologists and urological units. Statistical processing was done with SPSS for Windows 5.0. RESULTS: In the last 5 years, 501 urologists were licensed at an average age of 37 years; 535 urological units exist; 2,332 doctors practice urology (2,235 males and 97 females) for a ratio of 1 urologist to 24,500 inhabitants. By comparing the 'entrance' forecast with the potential 'exit', we can hypothesize an annual excess of 80 units. There is no significant correlation between the number of urologists in each structure and the number of inhabitants for each urologist. CONCLUSIONS: The present government programme does not take into account continual changes in employment and many other variables when defining the actual need for specialists. For valid predictions, the data we obtain must be updated for at least 5 years. PMID- 11054035 TI - Juxtaglomerular cell tumor of the kidney treated with nephron-sparing surgery. AB - A case of juxtaglomerular cell tumor of the right kidney is reported. A 30-year old woman visited us with a complaint of headaches. Severe hypertension and an elevated level of plasma renin activity was seen at the initial evaluation. Computerized tomographic angiography revealed tumor vessels in the low-density mass in the right kidney. The preoperative diagnosis was renin-secreting tumor of the kidney, and nephron-sparing surgery was performed. The pathological findings showed a juxtaglomerular cell tumor. Postoperatively, prompt normalization of blood pressure and a reduced plasma renin activity level were observed. PMID- 11054036 TI - Renomedullary interstitial cell tumor. AB - Renomedullary interstitial cell tumor was first introduced by Lerman and co workers. These lesions have been referred to as medullary fibromas. The ultrastructural studies showed that the spindle cells throughout the stroma have the features of medullary interstitial cells. These benign tumors with specific histology are rare since they are incidental findings at autopsies. We report here a case of renomedullary interstitial cell tumor in a 55-year-old woman. PMID- 11054037 TI - Malignant neurofibroma of the urinary bladder. AB - Malignant neurofibroma of the urinary bladder is a very rare entity and usually associated with von Recklinghausen's disease. We present the first case of sporadic malignant neurofibroma of the urinary bladder and a review of the literature. PMID- 11054038 TI - Carcinosarcoma in a bladder diverticulum. A case report and literature review. AB - Carcinosarcoma of the bladder is a rare neoplasm composed of a mixture of malignant epithelial and mesenchymal components. The presentation and clinical course of an infiltrating carcinosarcoma located in a bladder diverticulum coexisting with prostatic adenocarcinoma in a 69-year-old man are described. We believe this to be the fifth report of true carcinosarcoma appearing in a bladder diverticulum. PMID- 11054039 TI - Pheochromocytoma of the urinary bladder presenting only with macroscopic hematuria. AB - Pheochromocytoma of the bladder is an unusual tumor that typically presents with hypertensive crises related to micturition. We describe a case of bladder pheochromocytoma in a 42-year-old female in which macroscopic hematuria was the only alarming symptom. The diagnostic and operative issues of this type of tumor are discussed, along with the challenging treatment option of transurethral resection. Diagnosis, treatment and follow-up trends of this rare tumor are reviewed. PMID- 11054040 TI - Bladder pheochromocytoma: a 3-year follow-up after transurethral resection (TURB). AB - We report a case of a 75-year-old female with pheochromocytoma of the bladder. Clinical evaluation included ultrasonography, intravenous pyelography, CT scan, I MIBG scintiscan. A transurethral resection was performed for a exophytic tumor of 2 cm diameter. The histological result indicated the diagnosis of bladder pheochromocytoma. Three years later the patient remains disease free. Preoperative diagnosis is established by determination of blood and urine levels of catecholamines and their metabolites is a nonspecific diagnostic tool. The sensitivity of CT scan is 82%. Iodine-methyliodobenzylguanidine (I-MIBG), used by scintiscanning, specifically accumulates in pheochromocytomas. Life-long follow up is necessary to diagnose late recurrences. PMID- 11054041 TI - Ureteral displacement due to a migrated intrauterine contraceptive device. AB - Perforation of the uterus is rare but potentially fatal. During puerperium when the uterus is small and its wall is thin, the risk of perforation increases. We report a rare complication from an intrauterine contraceptive device (IUD) which caused deviation of the right ureter in a 31-year-old woman who presented with complaints of insomnia and abdominal pain. Our case shows that perforation of the uterus by an IUD can cause a silent urological complication. The possibility of ureteral involvement and displacement should be kept in mind in a woman in whom a missing IUD is encountered. PMID- 11054042 TI - PCR detects HCV RNA in a plasma pool contaminated by a single preseroconversion donation of genotype 5a. AB - BACKGROUND AND OBJECTIVES: To determine the prevalence of HCV-RNA-positive plasma pools in Belgium, to validate our PCR method and to increase the safety of the released blood products. MATERIALS AND METHODS: Plasma pools consisting each of about 5,000 donations from Belgian unpaid volunteer blood donors were analysed by PCR for the presence of HCV RNA. Two different extraction methods were compared and validated. RESULTS: Two out of 367 plasma pools were found to be HCV RNA positive and were discarded. For one of these two pools, the look-back procedure identified an anti-HCV-negative contaminated donation. The HCV genotype of both the contaminated pool and the donation was 5a, a genotype rare in Europe. The viral load of the preseroconverted donation was 2.9 x 10(7) gEq/ml according to the bDNA method. CONCLUSION: In the case of plasma derivatives, various important steps are already included to increase safety. Nucleic acid testing of manufacturing plasma pools ensures that viral load in the starting material is as low as possible. PMID- 11054043 TI - Retrospective analysis of blood transfusion recipients: evidence for post transfusion hepatitis E. AB - BACKGROUND AND OBJECTIVES: Of 200 volunteer blood donors we had screened earlier for hepatitis E virus (HEV) RNA, using reverse-transcription polymerase chain reaction, 3 were positive, raising the possibility of transfusion-associated hepatitis E in areas endemic for this virus. This retrospective study was to reassess the extent of post-transfusion hepatitis E among transfusion recipients, investigated in 1982. MATERIALS AND METHODS: We re-evaluated 56 recipients followed biweekly for 3 months after transfusion. The controls were 51 normal, healthy persons who gave blood at a 2-month interval, as well as 412 blood donors from whom blood was taken once in 1982. RESULTS: Of the 56 transfusion recipients, 19 were positive for IgG antibodies against HEV (anti-HEV) in the pretransfusion sample. Two of the 37 IgG anti-HEV-negative recipients seroconverted to IgM and IgG anti-HEV 5 and 4 weeks after transfusion, 1 with raised serum alanine aminotransferase levels. None showed symptoms of hepatitis. Attempts to detect HEV RNA in transfused blood, from aliquot units stored at -20 degrees C for over 17 years, were not successful. Of the controls, 17 out of 51 were IgG anti-HEV positive in the initial sample itself. None of the 34 IgG anti HEV-negative controls seroconverted during the 2-month follow-up. Of the blood donors, 154 out of 412 were IgG anti-HEV positive. None of the 412 donors had circulating IgM anti-HEV antibodies. A significantly higher (p<0.03) proportion of susceptible transfusion recipients were IgM anti-HEV positive as compared with susceptible blood donors. CONCLUSION: The results suggest that, in countries where HEV is endemic, the transmission of hepatitis E may be associated with blood transfusion. PMID- 11054044 TI - Gamma-ray-irradiated red blood cells stored in mannitol-adenine-phosphate medium: rheological evaluation and susceptibility to oxidative stress. AB - BACKGROUND AND OBJECTIVES: To evaluate the rheological properties and the oxidative susceptibility of gamma-ray-irradiated red blood cells (RBCs). MATERIALS AND METHODS: RBCs in mannitol-adenine-phosphate (MAP) medium were irradiated with 35 Gy and stored at 4 degrees C for 4 weeks. The deformability of the RBCs was examined under shear flow in relation to the morphological and biochemical changes. The RBCs were further exposed to 1 mM FeSO(4) and 5 mM ascorbate to examine the oxidative susceptibility. RESULTS: The RBC deformability was decreased during storage, and the impairment was further enhanced by the irradiation, which promoted cell shrinkage and intracellular hemoglobin condensation accompanying potassium loss. Lipid peroxidation and protein aggregation of the RBC membrane as well as echinocytosis were not enhanced by the irradiation. The exposure to free iron did not stimulate the oxidation of the irradiated RBC membrane. CONCLUSION: The decreased deformability of gamma-ray irradiated RBCs in MAP medium was mainly induced by dehydration due to potassium loss, and the membrane lipids and proteins were stably preserved against oxidative stress. PMID- 11054045 TI - Variable leukocyte composition of red blood cell concentrates prepared in top bottom systems: possible implications for pre-transplant blood transfusion. AB - BACKGROUND AND OBJECTIVES: The beneficial effect of blood transfusion on kidney graft survival requires the presence of leukocytes in the transfusate, but a minimal dose has not been defined, nor has the role of individual leukocyte subsets been investigated. In the Netherlands, a standard pre-transplant blood transfusion consists of a buffy coat (BC)-depleted red blood cell concentrate (RBCC) containing a maximum of 1.2x10(9) residual leukocytes per unit. However, leukocyte subset composition is not standardized. MATERIALS AND METHODS: Using FACS analysis, this study compared the residual leukocyte composition of RBCCs produced by Compomat((R)) and Optipress((R)), two currently used top-bottom systems. RESULTS: While the total leukocyte content of the RBCCs was equivalent in both press types (0.5x10(9)), the percentage of mononuclear cells (lymphocytes and monocytes) was significantly higher in the Compomat as compared with the Optipress system (p < 0. 0001), resulting in significantly higher numbers of transfused T cells, B cells, HLA-DR-positive cells, NK cells and stem cells. CONCLUSIONS: The leukocyte composition of a pre-transplant blood transfusion depends on the BC depletion method used; this might differentially affect the tolerizing or immunizing potential of a pre-transplant blood transfusion. PMID- 11054046 TI - Low leukocyte contamination without filtration by preparation of platelet concentrates in cylindrical bags with the buffy-coat method. AB - BACKGROUND AND OBJECTIVES: The buffy-coat (BC) method for platelet concentrate (PC) preparation was modified in order to obtain leukodepleted PCs from single BCs without filtration. MATERIALS AND METHODS: BCs were centrifuged in cylindrical BC bags and the optimal centrifugation conditions and optimal hematocrit were determined. RESULTS: With optimal conditions, a tenfold lower leukocyte contamination was obtained compared with the conventionally shaped, wide BC bag (0.3 +/- 0.19 versus 3.0 +/- 1.71 x 10(6) leukocytes per unit; 85-ml BCs). The platelet yield obtained with the cylindrical bag did not differ significantly from the yield obtained with the conventional bag (56 +/- 16.4 versus 61 +/- 15 x 10(9) platelets per PC). Furthermore, when PCs were prepared from 100-ml BCs in cylindrical bags, a leukocyte contamination of 0.2 +/- 0.11 x 10(6) and a platelet content of 61 +/- 13.5 x 10(9) per PC were obtained. CONCLUSION: The use of cylindrical BC bags reduced the leukocyte contamination in PCs to a level required for leuko-depletion without affecting platelet recovery. PMID- 11054047 TI - Automated collection of peripheral blood stem cells with the COBE spectra for autotransplantation. AB - BACKGROUND: A convenient, effective and safe peripheral blood stem cell (PBSC) apheresis procedure is desirable to cope with the increasing requirements for PBSC collections. We performed PBSC harvesting with the novel COBE Spectra AutoPBSC(TM) system using the default software configuration recommended by the manufacturer. We analyzed collection parameters and clinical efficiency of harvested autografts following high-dose chemotherapy (HDCT). PATIENTS AND METHODS: Eighty-one patients underwent 102 harvests after standard chemotherapy plus filgrastim (5-10 microg/kg/day) to obtain a target of >/=2.5 x 10(6) CD34+ cells/kg for autologous blood stem cell transplantation. Conventional-volume leukaphereses (median: 11 liters) were performed using the manufacturer's standard software default regarding inlet flow, harvest/chase volume (3/7 ml) and number of collection cycles. The ratio of ACD-A to whole blood was initially set at 1:12 (56 collections), later at 1:10 (46 aphereses). RESULTS: With respect to preapheresis counts of 93 (9-876) CD34+ cells/microl, 69 patients (85.2%) achieved >/=2.5 x 10(6) CD34+ cells/kg by the first apheresis. PBSC products contained medians of 5.0 x 10(6) (0.7-77.3) CD34+ cells/kg and 13.8 x 10(4) (2.3 105.0) CFU-GM/kg. A preapheresis count of >/=40 CD34+ cells/microl predicted a single-apheresis yield of >/=2.5 x 10(6) CD34+ cells/kg. Apheresis products showed a high mononuclear cell (MNC) purity of >/=89%. The median overall collection efficiency of CD34+ cells (CD34-CE) was 42.6% (12.2-87.4). The CD34-CE decreased significantly with increasing numbers of circulating CD34+ cells: 52.5% at CD34+ cells <40/microl versus 41.0% at CD34+ cells >/=40/microl (p 0.5 x 10(3)/microl, 10 (8-13) days for WBC >1.0 x 10(3)/microl and 11 (8-17) days for platelets >20 x 10(3)/microl. CONCLUSIONS: As a result of efficient PBSC mobilization, a single conventional volume leukapheresis with the COBE Spectra AutoPBSC system resulted in hematopoietic autografts with >/=2.5 x 10(6) CD34+ cells/kg in 85% of patients. Following the standard PBSC apheresis recommendations of the manufacturer, the AutoPBSC system assures PBSC products with a high MNC purity and a moderate CD34 CE that declines significantly at increasing levels of circulating CD34+ cells. Leukaphereses performed at an ACD-A to whole blood ratio of 1:10 should run without coagulation problems. PMID- 11054048 TI - A competitive enzyme-linked immunoassay using erythrocytes fixed to microtitre plates for anti-D quantitation in immunoglobulin products. AB - BACKGROUND AND OBJECTIVES: The batch control of anti-D immunoglobulin for prevention of haemolytic disease of the newborn necessitates assessment of its potency. Anti-D quantitation is usually performed using automated haemagglutination methodology although this can only be carried out in specialist centres. The aim of this study was to develop a simple and robust assay for anti D quantitation. METHODS: We developed a competitive enzyme-linked immunoassay (EIA) in which unlabelled anti-D immunoglobulin and a biotinylated monoclonal anti-D compete for red cell binding. Binding of biotinylated anti-D is detected using an alkaline-phosphatase-labelled avidin preparation. The assay is conveniently carried out using erythrocytes fixed to microtitre plates. RESULTS: The competitive EIA was specific for anti-D activity, highly reproducible and showed good correlation with manufacturers' potency estimates using automated haemagglutination. The assay was quick and simple to perform using freshly prepared or stored plates, and the biotinylated monoclonal anti-D could be lyophilized in ampoules for distribution as a standardized reagent. CONCLUSIONS: The competitive EIA described can be used for the specific quantitation of anti-D and provides a robust alternative method to automated haemagglutination. PMID- 11054049 TI - HLA-C expression on platelets: studies with an HLA-Cw1-specific human monoclonal antibody. AB - BACKGROUND AND OBJECTIVES: The expression of HLA-C on the surface of platelets is rarely studied due to the lack of proper alloantisera. We addressed this question using an IgM human monoclonal antibody directed against HLA-Cw1 (VP6G3). MATERIAL AND METHODS: Both flow cytometry and complement dependent cytotoxicity studies were used in the current analysis. RESULTS: The expression of the HLA-Cw1 antigen on platelets is lower than on peripheral blood lymphocytes as shown by flow cytometry. Variation in expression levels between individuals is also observed. Using this antibody in a complement-dependent cytotoxicity assay, we did not observe lysis using platelets as targets, whereas peripheral blood lymphocytes of the same blood donors were adequately lysed. CONCLUSIONS: These results confirm that platelets indeed express HLA-C. Furthermore, the results support the insignificant role of HLA-C in immunological platelet refractoriness. PMID- 11054050 TI - Transfusion-related acute lung injury associated with interdonor incompatibility for the neutrophil-specific antigen HNA-1a. AB - BACKGROUND AND OBJECTIVES: A patient transfused with two pooled platelet concentrates became breathless. Bilateral infiltrates were seen on chest X-ray. A diagnosis of transfusion-related acute lung injury (TRALI) was made. The patient received 100% oxygen and recovered after 5 days. MATERIALS AND METHODS: Antibody screening, cross-matching for granulocyte and lymphocyte antibodies and typing for granulocyte antigens was undertaken. RESULTS: The patient typed as HNA-1b/HNA 1b. Granulocyte and lymphocyte antibodies were not detected in the patient's serum or in any of the donor sera by cross-match. In antibody screening against typed panel granulocytes, complement-fixing anti-HNA-1a IgM antibodies were detected in the serum of one female donor. Two of the other donors who contributed to the pooled platelet concentrate containing the HNA-1a IgM antibodies typed as HNA-1a/HNA-1b. CONCLUSION: Anti-HNA-1a IgM antibodies may have formed immune complexes with white cell fragments or soluble FcgammaRIII from HNA-1a+ donors in the pooled platelet concentrate and initiated TRALI. PMID- 11054051 TI - New RhD(IVb) identified in Japanese. PMID- 11054052 TI - Parvovirus B19 in idiopathic thrombocytopenic purpura. PMID- 11054053 TI - Antibody to human foamy virus not detected in individuals treated with blood products or in blood donors. PMID- 11054054 TI - French national tissue banking activity. PMID- 11054055 TI - Guidelines for mitochondrial DNA typing. DNA Commission of the International Society for Forensic Genetics. PMID- 11054056 TI - Images in haematology. Osteitis fibrosa cystica generalisata with adult T-cell leukaemia: a case report. PMID- 11054057 TI - The history of blood transfusion. PMID- 11054058 TI - Dyskeratosis congenita in all its forms. PMID- 11054059 TI - The clinical and cellular pharmacology of vincristine, corticosteroids, L asparaginase, anthracyclines and cyclophosphamide in relation to childhood acute lymphoblastic leukaemia. PMID- 11054060 TI - Classification of ex vivo methotrexate resistance in acute lymphoblastic and myeloid leukaemia. PMID- 11054061 TI - Investigation of 9-O-acetylated sialoglycoconjugates in childhood acute lymphoblastic leukaemia. PMID- 11054062 TI - In vitro drug resistance profiles of adult versus childhood acute lymphoblastic leukaemia. AB - The difference in the current cure rates between adult and childhood acute lymphoblastic leukaemia (ALL) may be caused by differences in drug resistance. Earlier studies showed that in vitro cellular drug resistance is a strong independent adverse risk factor in childhood ALL. Knowledge about cellular drug resistance in adult ALL is still limited. The present study compared the in vitro drug resistance profiles of 23 adult ALL patients with that of 395 childhood ALL patients. The lymphoblasts were tested by the MTT assay. The group of adult ALL samples was significantly more resistant to cytosine arabinoside, L-asparaginase, daunorubicin, dexamethasone and prednisolone. The resistance ratio (RR) was highest for prednisolone (31.7-fold) followed by dexamethasone (6.9-fold), L asparaginase (6. 1-fold), cytosine arabinoside (2.9-fold), daunorubicin (2.5 fold) and vincristine (2.2-fold). Lymphoblasts from adult patients were not more resistant to mercaptopurine, thioguanine, 4-HOO-ifosfamide, mitoxantrone and teniposide. There were no significant differences in drug resistance between adult T-cell (T-) ALL (n = 11) and adult common/pre-B-cell (B-) ALL (n = 10). Additionally, adult T-ALL did not differ from childhood T-ALL (n = 69). There were significant differences between adult common/pre-B-ALL and childhood common/pre-B-ALL (n = 310) for prednisolone (RR = 302, P = 0.008), dexamethasone (RR = 20.9, P = 0.017) and daunorubicin (RR = 2.7, P = 0.009). Lymphoblasts from adults proved to be relatively resistant to drugs commonly used in therapy. This might contribute to the difference in outcome between children and adults with ALL. PMID- 11054063 TI - Activation of an ataxia telangiectasia mutation-dependent intra-S-phase checkpoint by anti-tumour drugs in HL-60 and human lymphoblastoid cells. AB - In yeast cells, the intra-S-phase checkpoint slows down the rate of DNA replication in response to DNA damage. Here we showed that a similar checkpoint mechanism is present and activated by anti-tumour drugs in HL-60 and Epstein-Barr virus (EBV)-transformed human lymphoblastoid cells. Using bromodeoxyuridine (BrdU) pulse labelling combined with two-dimensional flow cytometric analysis, we clearly visualized the cell-cycle progression of the BrdU-positive population (cells originally belonging to the S phase) and detected even subtle changes in S phase progression induced by mild drug treatment conditions free of apoptosis. The DNA topoisomerase II inhibitors, doxorubicin and etoposide (250 nmol/l and 400 nmol/l, respectively, for 8 h), retained the BrdU-positive HL-60 cells in the latter half of S and G2/M positions, and the pyrimidine analogue anti-metabolite, cytosine beta-D-arabinofuranose (Ara-C; 50 nmol/l), kept them in early-to-late S phase after 8 h of incubation. Because 10 micromol/l of caffeine added 2 h later attenuated the S-phase retardation by these drugs in HL-60 cells, slowing of the S-phase progression should be actively regulated. Furthermore, two ataxia telangiectasia (AT)-derived lymphoblastoid cell lines were impaired in the doxorubicin-induced S-phase retardation, which indicated that the process is at least partially dependent on ataxia telangiectasia mutated (ATM) gene product. The inhibitory mechanism on S-phase progression elicited by anti-tumour drugs in HL-60 and lymphoblastoid cells may therefore correspond to the intra-S-phase checkpoint of the yeast cells. PMID- 11054064 TI - Detection of different Ikaros isoforms in human leukaemias using real-time quantitative polymerase chain reaction. AB - The Ikaros gene is an essential regulator in development and haematopoiesis. Dysregulated Ikaros gene expression participates in leukaemic processes, as evidenced in animal models, and by analyses of blast-cell populations from leukaemic patients. We used real-time quantitative polymerase chain reaction (PCR) to evaluate the relative abundance of several Ikaros transcript isoforms in a variety of leukaemic-cell samples. Total RNA was isolated from bone-marrow or blood-cell samples collected at diagnosis in children or adult patients, 18 of whom had acute myeloblastic leukaemia (AML), 61 of whom had acute lymphoblastic leukaemia (ALL) and 11 of whom had chronic myeloid leukaemia (CML). The ratio (Ik1 + Ik2)/(Ik1 + Ik2 + Ik4 + Ik7 + Ik8) ranged from 13.5% to 85% and was lower (P < 0. 05) in samples from patients with m-bcr-abl ALL. An alternative splicing resulting in the deletion of 30 nucleotides at the end of exon 6 was observed in leukaemic samples, and in normal thymus and bone marrow. Our results are consistent with previous reports and suggest that the pattern of expression of the different human Ikaros isoforms are not homogeneous among different subsets of leukaemias. PMID- 11054065 TI - T-cell prolymphocytic leukaemia: antigen receptor gene rearrangement and a novel mode of MTCP1 B1 activation. AB - T-cell prolymphocytic leukaemia (T-PLL) is a sporadic, mature T-cell disorder in which there is usually an aberrant T-cell receptor alpha (TCRA) rearrangement that activates the TCL1 or MTCP1-B1 oncogenes. As mutations of the Ataxia Telangiectasia (A-T) gene, ATM, are frequent in T-PLL and as ATM seems to act as a tumour suppressor through a mechanism involving V(D)J recombination, we examined V(D)J recombination in T-PLL. Using Southern blotting and the polymerase chain reaction, two of 60 TCRG coding joints were abnormal. In all cases, both TCRD alleles were deleted, IGH was germline, and patterns of TCRB and TCRA rearrangement were normal. However, in a case harbouring t(X;7)(q28;q35), we identified TCRB segment J beta 2.7 juxtaposed to MTCP1 exon 1. This is the first time that TCRB has been implicated in MTCP1 B1 activation. The structure of the breakpoint supports a model in which translocation activates a cryptic MTCP1 promoter. This analysis of V(D)J recombination is consistent with it being a variable that is independent of ATM in T-PLL. PMID- 11054066 TI - The incidences of trisomy 8, trisomy 9 and D20S108 deletion in polycythaemia vera: an analysis of blood granulocytes using interphase fluorescence in situ hybridization. AB - We have used interphase fluorescence in situ hybridization (IFISH) to detect trisomy 8, trisomy 9 and 20q deletion in circulating granulocytes from patients with polycythaemia vera (PV). Out of 64 PV patients, 15 (23%) exhibited an abnormality. Two patients had trisomy 9, three had trisomy 8 and 10 patients had hemizygous deletion of D20S108 (a locus in the 20q common deleted region). Aberrant nuclei ranged from 10% to 80% in these 15 cases. There was no correlation between the presence of a marker and sex, age, interval between presentation and IFISH analysis, neutrophil or platelet count or therapy. Conventional marrow cytogenetic karyotype results were available in 23 cases and there was concurrence between these and blood IFISH in 16 cases (13 normal and three with 20q/D20S108 deletion by both methods). Three patients with D20S108 deletion by IFISH were normal by previous marrow cytogenetic testing and four cases with 20q deletion by previous marrow cytogenetics had normal blood granulocytes according to IFISH. Thus, we confirm that trisomies 8 and 9 and deletion of 20q are diagnostically useful markers of PV. IFISH analysis of blood granulocytes is a practical method for detecting these markers, but as an adjunct to, not as a substitute for, conventional marrow cytogenetics. PMID- 11054067 TI - Cytogenetic and molecular diagnosis of chromosome 5 deletions in myelodysplasia. AB - Deletions of chromosome 5, del(5q), are frequently observed in myelodysplasia (MDS). We evaluated molecular detection of loss of heterozygosity (LOH) as a diagnostic method to detect del(5q) in a series of 60 MDS cases at a single institution. LOH was compared to cytogenetics on the same clinical specimen, resolving ambiguous cases by fluorescent in situ hybridization (FISH) and additional LOH. There was poor concordance between molecular and cytogenetic results, but most discrepancies could be resolved by FISH and additional LOH. Molecular analysis was of low sensitivity because most cases contained a relatively high proportion of cells without del(5q), but it was accurate, while cytogenetics overestimated the proportion of cells with del(5q) and failed to detect some cases with complex rearrangements. Minor clones were detected both by FISH and LOH. Overall, we found an incidence of 23% (14 of 60 cases) for del(5q) in MDS. The results also suggest that there is a high degree of genetic heterogeneity in the cellular population of MDS. Although del(5q) is common in MDS, it may not be present in all cells, leading to diagnostic challenges. PMID- 11054068 TI - Diagnostic utility of fluorescence in situ hybridization in mantle-cell lymphoma. AB - Mantle-cell lymphoma (MCL) has a poorer prognosis than other small B-cell lymphomas, thus a definitive diagnosis is essential. The t(11;14)(q13;q32) associated with MCL juxtaposes portions of CCND1 (11q13) and IGH (14q32), resulting in over-expression of cyclin D1. In this study, a highly sensitive two colour fluorescence in situ hybridization (FISH) method was developed to detect t(11;14)(q13;q32) in nuclei isolated from paraffin-embedded tissue. Twenty-three MCLs, 13 normal controls and nine small B-cell lymphomas other than MCL were studied by FISH. Each MCL had been previously investigated to detect genomic IGH CCND1 fusion by polymerase chain reaction (PCR) using DNA extracted from frozen tissue. The IGH-CCND1 fusion detection rate in the MCLs was 96% by FISH compared with 35% by PCR. By FISH, one MCL and three small B-cell lymphomas other than MCL harboured abnormalities involving only IGH. Less than 1% of cells showed false positive IGH-CCND1 fusion in normal specimens by FISH. Thus, this highly sensitive FISH assay is very useful in confirming the diagnosis of MCL, has wide applicability as it may be performed on both paraffin-embedded and fresh tissue, and may also facilitate detection of translocations involving these loci in tumours other than MCL. PMID- 11054069 TI - Primary myelosarcomas are associated with a high rate of relapse: report on 34 children from the acute myeloid leukaemia-Berlin-Frankfurt-Munster studies. AB - Primary myelosarcomas are rare manifestations of acute myeloid leukaemia (AML) that precede bone marrow involvement. Out of 744 children observed during the AML Berlin-Frankfurt-Munster (BFM) studies 87 and 93, 34 children presented with extramedullar myelosarcomas and no blasts (n = 21; 2.8%), or a low blast count (n = 13; 1.7%) in the bone marrow. Owing to the initially mild and variable symptoms, in some children (n = 12) diagnostic procedures were delayed and treatment intensity was reduced. At 0.65 +/- 0.13, the cumulative incidence of relapse was significantly higher than in other AML patients (0.28 +/- 0.02). The 5-year event-free survival was 0.19 +/- 0.08 (compared with 0.48 +/- 0.02 in AML BFM studies 87/93; P(log rank) < 0.03). Overall, 18 out of 34 patients died from disease (estimated 5 year survival 0.44 +/- 0.09 compared with 0.55 +/- 0.02 in the AML-BFM-studies 87/93; P(log rank) = 0.35, n.s.). An early diagnostic workup is needed in children with unusual skin lesions or tumours, considering myelosarcoma as a primary manifestation of AML. Intensive AML-specific chemotherapy is recommended soon after diagnosis. PMID- 11054070 TI - Novel BCR-ABL transcript containing an intronic sequence insert in a patient with Philadelphia-positive acute lymphoblastic leukaemia. AB - In a patient with Philadelphia chromosome-positive acute lymphoblastic leukaemia (ALL), a novel variant of the chimaeric BCR-ABL mRNA transcript was detected by reverse transcription polymerase chain reaction (RT-PCR). Sequencing revealed the novel transcript to be a chimaeric mRNA produced by fusion of the BCR exon 14 (b3) to the ABL exon a2 with a 49-base pair (bp) insertion of an ABL intron 1b sequence between them. The insertion of the 49 bp introduced a stop codon. These data show that this variant of the chimaeric mRNA would not be translated into the p210 BCR-ABL protein. This could be one of the explanations as to why clinically the patient has responded well to therapy and continues to follow a mild clinical course. PMID- 11054071 TI - Acute leukaemia in acromegaly patients. AB - Acromegaly patients are known to have an increased risk of malignancies, especially colonic adenocarcinoma. This may be as a result of the growth stimulating effect of growth hormone (GH). The clustering of leukaemia in children treated with GH has also caused concern. There have been a few reports of leukaemia in acromegaly patients. We report two patients with acute lymphoblastic leukaemia and one patient with acute myeloid leukaemia among 106 acromegaly patients treated over a 15-year period. Two of the cases received radiotherapy as part of their treatment. Adjusted for age and follow-up years, the incidence of leukaemia in this cohort is significantly higher than the general population. The incidence is also higher than would be expected as a result of radiotherapy alone, suggesting that GH may play a synergistic role. PMID- 11054072 TI - Sight-threatening varicella zoster virus infection after fludarabine treatment. AB - Varicella zoster virus (VZV) infection involving the posterior segment of the eye after fludarabine treatment has not previously been described. Two patients, who had completed fludarabine treatment 3 and 18 months previously, presented with visual loss that had been preceded by a recent history of cutaneous zoster. The use of the polymerase chain reaction (PCR) for VZV DNA from ocular specimens allowed rapid confirmation of clinical diagnosis and treatment with a good outcome in one patient. With the increasing use of fludarabine and other purine analogues, an awareness of such complications is important because of their potentially sight-threatening consequences. PMID- 11054073 TI - Involvement of natural killer cells in patients with myelodysplastic syndrome carrying monosomy 7 revealed by the application of fluorescence in situ hybridization to cells collected by means of fluorescence-activated cell sorting. AB - Monosomy 7 is the most frequent chromosome abnormality among patients with secondary myelodysplastic syndrome (MDS). We used fluorescence in situ hybridization (FISH) and fluorescence-activated cell sorting (FACS) in order to clarify the lineage involvement. Four patients, three with de novo MDS and one with secondary MDS, were enrolled in this study. Monosomy 7 was observed in pluripotent stem cells (CD34(+)Thy-1(+)), and in B (CD34(+)CD19(+)) and T/natural killer (NK) progenitors (CD34(+)CD7(+)). The number of abnormal cells of B (CD19(+)) and T (CD3(+)) cells was below the cut-off value, but approximately 60% of the NK cells (CD3-CD56(+)) contained monosomy 7 in three of the patients. PMID- 11054074 TI - Nephrotic syndrome caused by protein thrombi in glomerulocapillary lumen in Waldenstrom's macroglobulinaemia. AB - Waldenstrom's macroglobulinaemia (WM) is described as a disorder of plasmacytoid lymphocytes. The renal complications of WM are less common and severe than those of multiple myeloma. We present a case of WM complicated by nephrotic syndrome. A biopsy specimen of the kidney revealed the intraglomerular thrombi of immunoglobulin M paraprotein. Corticosteroid pulse therapy and plasmapheresis were effective in improving proteinuria and reducing protein thrombi. The nephrotic syndrome caused by protein thrombi in WM may be reversible, at least in its early stage. PMID- 11054075 TI - Long-term remission after intensive chemotherapy in advanced myelodysplastic syndromes is generally associated with restoration of polyclonal haemopoiesis. AB - The clonality of peripheral blood cells was assessed in eight female patients with myelodysplastic syndrome (MDS) by means of the human androgen receptor gene based assay (HUMARA). The patients were in complete remission for a median follow up time of 83 months after intensive chemotherapy. X-chromosome inactivation patterns (XCIPs) indicated polyclonal haemopoiesis in five patients. Two patients had skewed lyonization (i.e. unbalanced XCIPs in both granulocytes and T cells) and one patient presented monoclonal granulocytes together with polyclonal T cells. We conclude that long-term remission in MDS following intensive chemotherapy is usually associated with polyclonal haemopoiesis. PMID- 11054076 TI - Feasibility and recent improvement of autologous stem cell transplantation for acute myelocytic leukaemia in patients over 60 years of age: importance of the source of stem cells. AB - A total of 193 patients with acute myelocytic leukaemia (AML) [147 in first complete remission (CR1)], ranging from 60 years to 75 years of age (median 63 years), were autografted between January 1984 and December 1998. The source of stem cells was peripheral blood (PB) in 128 patients, bone marrow in 51 patients and a combination of both in 14 patients. Total body irradiation (TBI) was used in 34 cases. Ninety-seven per cent of patients had successful engraftment of neutrophils on day 15 (range days 7-71) and of platelets on day 30 (range days 9 894). In patients autografted in CR1, the transplant-related mortality (TRM) was 15 +/- 4%, the relapse incidence (RI) was 58 +/- 5%, the leukaemia-free survival (LFS) was 36 +/- 5% and the overall survival was 47 +/- 5% at 3 years. The source and dose of stem cells were studied in particular; in patients transplanted in CR1, the RI was 44 +/- 11% in those receiving marrow compared with 63 +/- 6% in those receiving PB (P = 0.04). Patients autografted in CR1 who received higher granulocyte-macrophage colony-forming units (CFU-GM) doses (above the median) had a lower RI (47 +/- 11% vs. 79 +/- 9%, P = 0.009). There was a significant improvement in patients transplanted after March 1996; for those in CR1, the RI was 41 +/- 8% vs. 65 +/- 6% (P = 0.01), the LFS was 53 +/- 8% vs. 28 +/- 5% (P = 0.01) and the overall survival was 72 +/- 7% vs. 36 +/- 6% (P = 0.02). By multivariate analyses, significant factors for the outcome were the date of transplant with recent improvement and the source of stem cells, with a lower RI for marrow. Autologous stem cell transplantation (ASCT) is a potential therapeutic approach in patients with AML over 60 years of age; results have improved recently. PMID- 11054077 TI - Cytomegalovirus colitis after autologous transplantation for multiple myeloma. AB - Bone marrow or stem cell transplantation is an established therapy for haematological malignancies. We report a cytomegalovirus (CMV) IgG +ve 56-year old patient who underwent autologous rescue with CD34(+) selected peripheral blood stem cells as part of consolidation therapy for multiple myeloma and subsequently developed CMV colitis. In contrast to infection secondary to human immunodeficiency virus (HIV), CMV colitis has not previously been described in this context. We discuss this case and issues arising from it related to the use of CD34+ selected stem cells for transplantation. PMID- 11054078 TI - Allogeneic haematopoietic transplantation for Richter's syndrome. AB - This study evaluated the efficacy of allogeneic bone marrow transplantation (AlloBMT) in eight patients who had chronic lymphocytic leukaemia (CLL) in Richter's transformation. Five patients were in resistant relapse and three others were in sensitive or untreated relapse. Three out of eight patients (38%) were alive and in remission at 14 months, 47 months and 67 months respectively. Two of these three patients, including one with an unrelated-donor transplant and a prior, unsuccessful autologous transplantation, received a non-myeloablative preparative regimen. These data suggest that, compared with conventional chemotherapy, AlloBMT improves the prognosis for patients with Richter's syndrome. Further study of a larger number of patients is needed to confirm these results. PMID- 11054079 TI - Two-step immunoablative treatment with autologous peripheral blood CD34(+) cell transplantation in an 8-year-old boy with autoimmune haemolytic anaemia. AB - Life-threatening haemolysis in children with autoimmune haemolytic anaemia (AIHA) occurs rarely. Many cases of severe autoimmune disease are currently treated with immunosuppressive high-dose chemotherapy and autograft. We report here a case of a child with severe AIHA who did not respond to conventional treatments, but was cured with an autologous peripheral blood CD34(+) cell transplantation. After d 16 post autograft, no further red cell transfusions were required. At 20 months post autograft, haematological complete remission persists. PMID- 11054080 TI - Retrovirus-mediated transfer of the herpes simplex virus thymidine kinase and enhanced green fluorescence protein genes in primary T lymphocytes. AB - The EGFP-tk retroviral vector, encoding enhanced green fluorescent protein (EGFP) and the herpes simplex virus thymidine kinase (HSV-tk) packaged in a Phoenix amphotropic cell line, was used to transduce healthy donor T lymphocytes. Infection yielded a mean of 41.8 +/- 9.3% SD (range 31.1-48.4%) EGFP-positive cells and a mean of 92 +/- 2% SD (range 90-94%) after cell sorting. EGFP expression remained stable for 30 d after infection. The entire gene transfer procedure had no significant effect on lymphocyte subsets and slightly reduced clonogenicity. Ganciclovir (gcv) treatment (1 microg/ml x 10 d) killed all EGFP positive cells in the transduced and transduced/sorted populations, but had no effect on untransduced controls. Our results show that primary T lymphocytes can be transduced using an EGFP-tk vector that yields a homogeneous infected population without affecting lymphocyte subsets, function and clonogenicity. PMID- 11054081 TI - Glycoprotein Ib/IX complex is the target in rifampicin-induced immune thrombocytopenia. AB - Thrombocytopenia is a major adverse effect of several drug treatments. Rifampicin has been recognized as a cause of immune thrombocytopenia during intermittent high-dose therapy. We characterized the antibody of a patient who presented with purpura and thrombocytopenia during treatment of tuberculosis with rifampicin. Drug-dependent binding of the antibody to platelets was demonstrated by flow cytometry. In a glycoprotein-specific immunoassay, the binding epitope of the IgG antibody was found in the glycoprotein Ib/IX complex, using four different monoclonal antibodies (mAbs) against various epitopes on the GPIb/IX complex, as well as mAbs against GPIIb/IIIa, GPIa/IIa and GPIV. By immunoprecipitation of biotin-labelled platelets, reactivity of the antibody with GPIb/IX was found only in the presence of the drug. These findings clearly demonstrate that rifampicin induces the formation of drug-dependent antibodies capable of causing thrombocytopenia. The binding site of the rifampicin-dependent antibody, located in the GPIb/IX complex, seems to be a favoured target for antibodies induced by different drugs. PMID- 11054082 TI - Defective platelet response to arachidonic acid and thromboxane A(2) in subjects with Pl(A2) polymorphism of beta(3) subunit (glycoprotein IIIa). AB - The membrane complex alpha(IIb)beta(3) is the major receptor for fibrinogen and is involved in platelet adhesion and aggregation. Evidence has been presented that the Pl(A2) allele of the beta(3) Pl(A1/A2) gene polymorphism might be an independent risk factor for coronary thrombosis, but the matter is still controversial. We investigated the relationship between this polymorphism and possible alterations of platelet functions in vitro. The platelet adhesion to fibrinogen-coated microplate wells and the aggregation induced by several different agonists were tested in 63 healthy volunteers, among them, 49 subjects with Pl(A1/A1) polymorphism, 12 subjects with Pl(A1/A2) polymorphism and two subjects with (PlA2/A2) polymorphism. Subjects with PlA1/A2 polymorphism or with Pl(A2/A2) polymorphism showed significantly lower platelet responses as compared with Pl(A1/A1) subjects when either arachidonic acid or the thromboxane A(2) analogue, U46619, were used as agonists. In resting condition and after thrombin or ADP stimulation, platelet function was normal in all the subjects. An increased sensitivity to the anti-aggregatory effect of acetylsalicylic acid was observed in platelets from subjects with the Pl(A2) allele. Finally, using a flow cytometric evaluation and determining the beta-thromboglobulin plasma levels, we did not find any evidence of a Pl(A2) platelet hyper-reactivity ex vivo. Our findings are not consistent with the hypothesis that the purported increase of cardiovascular risk in these subjects may be as a result of platelet hyperactivation. On the contrary, the Pl(A2) allele is associated with a platelet functional deficiency, specifically linked to the activation of the fibrinogen receptor by thromboxane A(2). PMID- 11054083 TI - Bernard-Soulier syndrome in a patient doubly heterozygous for two frameshift mutations in the glycoprotein ib alpha gene. AB - We report here the genetic basis of Bernard-Soulier syndrome in a compound heterozygote for two mutant glycoprotein (GP) Ib alpha alleles. One allele contained a novel four base-pair deletion (TGAG) that eliminated the last base of the codon for Ser39 (AGT) and the entire codon for Glu40 (GAG), causing a reading frame shift that yielded a stretch of 51 amino acids before a premature stop codon. The other allele also contained a frame-shift mutation, caused by deletion of the last two bases of the codon for Tyr492 (TAT). This allele produced a truncated glycoprotein Ib alpha that, although not expressed on the surface of the patient's platelets, was detectable in the plasma. The second allele has been identified previously by our group and other investigators as the cause of Bernard-Soulier syndrome in patients of northern European ancestry. This allele carried a haplotype identical to those of the previously reported cases, with the following polymorphic markers: two tandem repeats in the VNTR region, C at nucleotide -5 from the ATG start codon and a substitution of G for A in the third base for codon Arg342. These findings suggest that this particular Bernard Soulier mutation occurred once on the background of a rare haplotype and has spread throughout the northern European population. PMID- 11054084 TI - The central role of the P(2T) receptor in amplification of human platelet activation, aggregation, secretion and procoagulant activity. AB - Adenosine diphosphate (ADP) is an important platelet agonist and ADP released from platelet dense granules amplifies responses to other agonists. There are three known subtypes of ADP receptor on platelets: P2X(1), P2Y(1) and P(2T) receptors. Sustained ADP-induced aggregation requires co-activation of P2Y(1) and P(2T) receptors. AR-C69931MX, a selective P(2T) receptor antagonist and novel antithrombotic agent, was studied to characterize further the function of the P(2T) receptor. The roles of the P2Y(1) receptor and thromboxane A(2) were assessed using the selective P2Y(1) antagonist A2P5P and aspirin respectively. Aggregation was measured by whole blood single-platelet counting and platelet rich plasma turbidimetry, using hirudin anticoagulation. Dense granule release was estimated using ([14)C]-5-hydroxytryptamine (HT)-labelled platelets. Ca(2+) mobilization, P-selectin expression, Annexin V binding and microparticle formation were determined by flow cytometry. P(2T) receptor activation amplified ADP-induced aggregation initiated by the P2Y(1) receptor, as well as amplifying aggregation, secretion and procoagulant responses induced by other agonists, including U46619, thrombin receptor-activating peptide (TRAP) and collagen, independent of thromboxane A(2) synthesis, which played a more peripheral role. P(2T) receptor activation sustained elevated cytosolic Ca(2+) induced by other pathways. These studies indicate that the P(2T) receptor plays a central role in amplifying platelet responses and demonstrate the clinical potential of P(2T) receptor antagonists. PMID- 11054085 TI - Fibrinogen polymorphisms are not associated with the risk of myocardial infarction. AB - In the Study of Myocardial Infarctions Leiden, we investigated the prevalence of three polymorphisms in the alpha- and beta-fibrinogen genes among 560 patients with a myocardial infarction and 646 control subjects. Secondly, we studied the relationships between these polymorphisms and fibrinogen activity and antigen levels. The TaqI, HaeIII and BclI polymorphisms in the fibrinogen gene were not associated with myocardial infarction. As we found an association of the rare B2 allele with fibrinogen levels and a similar, but weak, effect for the rare H2 allele, we conclude that a genetic propensity to high fibrinogen levels does not affect the risk of myocardial infarction. This is evidence against a causal role for fibrinogen levels in the aetiology of myocardial infarction. PMID- 11054086 TI - Factor V Leiden: the venous thrombotic risk in thrombophilic families. AB - Factor V Leiden (FVL) leads to a sevenfold increased risk of venous thrombosis and is present in 50% of individuals from families referred because of unexplained familial thrombophilia. We assessed the association of FVL with venous thromboembolism (VTE) in 12 thrombophilic families of symptomatic probands with FVL in a retrospective follow-up study. We screened 182 first- and second degree relatives of the 12 unrelated propositi for the FVL mutation and the occurrence of VTE. The incidence rate of VTE in carriers of FVL (0.56%/year) was about six times the incidence for the Dutch population (0.1%/year). The incidence rate in non-carriers also appeared to be higher (0.15% per year). At the age of 50 years, the probability of not being affected by VTE was reduced to 75% for carriers and to 93% for non-carriers (P = 0.009). Identification of carriers of FV Leiden may be worthwhile in young symptomatic individuals and their relatives with a strong positive family history of venous thromboembolism or a history of recurrent venous thrombosis who may be at risk (e.g. pregnancy, use of oral contraceptives). After adjustment for prothrombin G20210A (present in two families), even higher thrombotic incidence rates were found in carriers and non carriers of FVL. This makes the presence of other unknown prothrombotic risk factors more probable in these families. PMID- 11054087 TI - Haemostatic screening and identification of zebrafish mutants with coagulation pathway defects: an approach to identifying novel haemostatic genes in man. AB - Zebrafish were used as a model to study haemostasis, a vertebrate function of paramount importance. A limitation of the zebrafish model is the difficulty in assaying small amounts of blood to detect coagulation mutants. We report the use of a rapid total coagulation activity (TCA) assay to screen for coagulation defects in individual adult zebrafish. We screened the TCA in 1000 gynogenetic half-tetrad diploids derived from 86 clutches. Each clutch was from a single F1 female offspring of males mutagenized with ethylnitrosourea (ENU). We found 30 50% defective zebrafish among six clutches, consistent with a heritable defect. The assay developed here provided a rapid screen to detect overall coagulation defects. However, because of the limited amounts of plasma, we could not detect defects in specific pathways. Therefore, a novel, ultra-sensitive kinetic method was developed to identify specific pathway defects. To test whether the kinetic assay could be used as a screening tool, 1500 Florida wild-type zebrafish pairs were analysed for naturally occurring coagulation defects. We detected 30 fish with extrinsic pathway defects, but with intact common and intrinsic pathways. We conclude that it is now possible to identify specific coagulation pathway defects in zebrafish. PMID- 11054088 TI - Contrasting effects of HSP72 expression on apoptosis in human umbilical vein endothelial cells and an angiogenic cell line, ECV304. AB - The effect of overexpression of heat shock protein (HSP)72 on apoptosis induced by different stimuli in human umbilical vein endothelial cells (HUVECs) and the angiogenic cell line, ECV304, was studied. Transient overexpression of HSP72 was achieved using an adenoviral vector (Advhsp72) and apoptosis was induced by heat shock, tumour necrosis factor (TNF)-alpha with cycloheximide (CHX), lipopolysaccharide (LPS) with TNF-alpha and verocytotoxin (VT). Apoptosis induced by heat shock was reduced by HSP72 expression. However, HSP72 expression in HUVECs increased apoptosis induced by TNF-alpha/CHX, LPS and VT measured by flow cytometric analysis of propidium iodide (PI)-stained permeabilized cells. In contrast, apoptosis in ECV304 induced by the same stimuli was reduced by HSP72 expression. No difference was seen in cells transduced with a control adenoviral vector expressing beta-galactosidase. These data imply that induction of HSP72 in cells modulates responses to apoptotic stimuli, but that the nature of the response varies with the cell type. However, it is clear that in situations where apoptosis may be part of a pathological process, HSP72 induction, for example by reperfusion injury, may exacerbate the process. PMID- 11054089 TI - Relationship between bleeding time, aspirin and the PlA1/A2 polymorphism of platelet glycoprotein IIIa. AB - A single nucleotide T to C transition of the gene encoding glycoprotein IIIa leads to a common diallelic polymorphism Leu-33-->Pro (PLA1/A2). We studied the relationship between the PlA1/A2 polymorphism and platelet function in 80 healthy men, aged 20-25 years. Before aspirin ingestion, bleeding time (BT) was shorter in carriers of the PlA2 than in carriers of the PlA1/A1 allele. At 4 h after ingestion of 300 mg of aspirin, BT became prolonged, and the intergroup difference was enhanced. In seven out of 26 PLA2 allele carriers, aspirin shortened BT on average by 30 s, compared with only one among 54 subjects with the PlA1/A1 genotype. Thus, BT both at baseline and after aspirin depends on the PlA1/A2 polymorphism of glycoprotein IIIa. Carriers of the PlA2 allele appear to be more resistant to the antithrombotic action of aspirin. PMID- 11054090 TI - Th1 and Th2 cytokines in a patient with Evans' syndrome and profound lymphopenia. AB - A case of Evans' syndrome with IgM deficiency and lymphopenia was studied before and after splenectomy. The lymphopenia was as a result of profound reduction of CD4 and CD8 cells. Study of cytokine secretion before splenectomy revealed a spontaneous Th1- and Th2-type cytokine production, and complete suppression of transforming growth factor (TGF)-beta. After splenectomy, the patient achieved clinical remission, the natural killer (NK) cell number increased and the pattern of cytokine production showed normalization of interleukin (IL)-2, IL-4, IL-10, TGF-beta and abolition of interferon (IFN)-gamma production. We conclude that splenectomy had a beneficial effect owing to an increase in NK cells and an associated increase in TGF-beta production. PMID- 11054091 TI - Hepatic iron concentration combined with long-term monitoring of serum ferritin to predict complications of iron overload in thalassaemia major. AB - Clinical complications of transfusional iron overload are still common in patients with thalassaemia major (TM) and it is not clear how best to monitor body iron stores during long-term follow-up to anticipate tissue damage. In this study, we have reviewed a group of 32 patients who underwent liver biopsy between 1984 and 1986. We developed a method of assessing the trend in serum ferritin (TSF) during long-term monitoring and compared this with mean serum ferritin (MSF) and initial liver iron (LI) concentration to determine whether, individually or in combination, they were accurate in predicting clinical outcome. LI levels were low (< 7 mg/g), medium (7-15 mg/g) and high (> 15 mg/g dry weight) in 15, 7 and 10 patients respectively. MSF was low (< 1500 microg/l), medium (1500-2500 microg/l) and high (> 2500 microg/l) in 10, 14 and 8 patients. TSF was low, medium and high risk in 9, 9 and 11 out of 29 evaluable patients. During a median follow-up of 13.6 years (range 2.3-14.8 years) after biopsy, nine patients died and an additional three patients developed heart failure. Hypothyroidism developed in five, hypoparathyroidism in four, and diabetes mellitus in seven patients. Cirrhosis developed in four of 10 evaluable patients. The clinical end-point of death or cardiac failure was significantly associated with increasing iron load using all three means of assessment. Although numbers were insufficient for statistical analysis, MSF or TSF were more closely associated with complications of iron overload than LI. There was no clear additional value in combining LI with MSF or TSF. The data show that quantitation of liver iron from a single liver biopsy has little value in long-term monitoring of iron stores. Most complications can be avoided if ferritin levels can be brought down to <1500 microg/l. PMID- 11054092 TI - Relationship between TNF-alpha and iron metabolism in differentiating human monocytic THP-1 cells. AB - The human monocytic cell line THP-1 differentiates along the macrophage line after phorbol-12-myristate-13-acetate (PMA) supplementation and can be stimulated to secrete tumour necrosis factor alpha (TNF-alpha) by interferon gamma (IFN gamma) addition. We found that, in the early stage of differentiation (1-48 h), PMA induction elicited an upregulation of intracellular H ferritin and H ferritin binding sites and a downregulation of transferrin receptor. In addition, we found that iron administration to PMA-differentiating cells induced the expression of TNF-alpha mRNA and TNF-alpha secretion to levels even higher than those induced by IFN-gamma alone. The iron chelator desferrioxamine showed the opposite effect and reduced TNF-alpha release. In contrast, preincubation of the cells with iron before PMA induction resulted in a decrease of the TNF-alpha secretion induced by IFN-gamma, whereas the opposite was true after preincubation with desferrioxamine. The data support a co-ordinate interaction between iron and TNF alpha in monocyte macrophages, with an iron-mediated upregulation of TNF-alpha in the early phase of differentiation and an iron-mediated inhibition at later stages. This complex relationship has to be considered in evaluating the effects of iron on inflammation. PMID- 11054093 TI - Chelator-induced iron excretion in iron-overloaded marmosets. AB - In order to test new orally active iron chelators in a predictive way, a primate model has been developed. This model makes use of the marmoset monkey (Callithrix jacchus) and its overall design is similar to a previously reported monkey model. However, this new model enables a higher compound throughput and requires lower amounts of test compound because the animals are much easier to handle and have much lower body weights. The marmosets were iron-overloaded by three intraperitoneal injections of iron (III) hydroxide polyisomaltose. For the iron balance studies, the animals were kept in metabolic cages and were maintained on a low-iron diet in order to reduce faecal background. After compound administration, the excretion of iron in urine and faeces was followed for 2 d. A series of well-known chelators was tested for validation of the model. In particular, comparison of the iron-clearing properties of DFO, L1, CP94 and HBED in marmosets and humans demonstrated the predictive value of the model and justify our expectation that if iron chelators such as CGP65015, ICL670A and CGP75254A are active in marmosets, they will be active in humans as well. PMID- 11054094 TI - A new PKLR gene mutation in the R-type promoter region affects the gene transcription causing pyruvate kinase deficiency. AB - Mutations in the PKLR gene responsible for pyruvate kinase (PK)-deficient anaemia are mainly located in the coding regions: 11 are in the splicing sites and, recently, three mutations have been described in the promoter region. We now report a novel point mutation A-->G on nucleotide 72, upstream from the initiation codon of the PKLR gene, in four Portuguese PK-deficient patients. This new regulatory mutation occurs within the most proximal of the four GATA motifs (GATA-A element) in the R-type promoter region. In two patients who were homozygous for this mutation, a semiquantitative reverse transcription polymerase chain reaction (PCR) procedure was used to evaluate the amount of R-PK mRNA transcript in the reticulocytes. The mRNA level was about five times lower than in normal controls, demonstrating that the PKLR gene transcription is severely affected, most probably because the -72A-->G point mutation disables the binding of the erythroid transcription factor GATA-1 to the GATA-A element. Supporting these data, the two patients homozygous for the -72A-->G mutation had severe haemolytic anaemia and were transfusion dependent until splenectomy. Two other patients who were compound heterozygous for this mutation and the previously described missense mutation 1456C-->T had a mild condition. PMID- 11054095 TI - Short report: erythrocyte membranes from a patient with congenital dyserythropoietic anaemia type I (CDA-I) show identical, although less pronounced, glycoconjugate abnormalities to those from patients with CDA-II (HEMPAS). AB - Congenital dyserythropoietic anaemias (CDAs) are rare hereditary disorders characterized by ineffective erythropoiesis and multinuclearity of erythroblasts. Three main types of the disease have been described. Glycoconjugate abnormalities in erythrocyte membrane glycoconjugates, consisting of hypoglycosylation of band 3 and accumulation of certain glycosphingolipids including lactotriaosylceramide, neolactotriaosylceramide and polyglycosylceramides, have been described only in patients with CDA type II (CDA-II). We report on identical, although less pronounced, abnormalities in erythrocyte glycoconjugates from a patient with CDA I. A low degree of hypoglycosylation of band 3 in our patient with CDA-I suggests that hypoglycosylation is not a cause, but, most probably, a consequence of dyserythropoiesis. PMID- 11054096 TI - Childhood essential thrombocythaemia without evidence of myeloproliferation: how many investigations should be done? PMID- 11054097 TI - Rituximab salvage following relapse after allogeneic bone marrow transplantation for non-Hodgkin's lymphoma. PMID- 11054098 TI - Antenatal testing for haemoglobinopathies. PMID- 11054099 TI - Reply to telfer et al PMID- 11054100 TI - Transfection of receptor tyrosine kinase axl into human haematopoietic cells does not cause spontaneous cellular aggregation. PMID- 11054101 TI - Preretinal neovascularization in South-East Asian ovalocytosis. PMID- 11054102 TI - Amphioxus alcohol dehydrogenase is a class 3 form of single type and of structural conservation but with unique developmental expression. AB - The coding region of amphioxus alcohol dehydrogenase class 3 (ADH3) has been characterized from two species, Branchiostoma lanceolatum and Branchiostoma floridae. The species variants have residue differences at positions that result in only marginal functional distinctions. Activity measurements show a class 3 glutathione-dependent formaldehyde dehydrogenase, with kcat/Km values about threefold those of the human class 3 ADH enzyme. Only a single ADH3 form is identified in each of the two amphioxus species, and no ethanol activity ascribed to other classes is detectable, supporting the conclusion that evolution of ethanol-active ADH classes by gene duplications occurred at early vertebrate radiation after the formation of the amphioxus lineage. Similarly, Southern blot analysis indicated that amphioxus ADH3 is encoded by a single gene present in the methylated fraction of the amphioxus genome and northern blots revealed a single 1.4-kb transcript. In situ experiments showed that amphioxus Adh3 expression is restricted to particular cell types in the embryos. Transcripts were first evident at the neurula stage and then located at the larval ventral region, in the intestinal epithelium. This tissue-specific pattern contrasts with the ubiquitous Adh3 expression in mammals. PMID- 11054103 TI - Characterization of the internal motions of a chimeric protein by 13C NMR highlights the important dynamic consequences of the engineering on a millisecond time scale. AB - By transferring the central curaremimetic beta hairpin of the snake toxin alpha into the scaffold of the scorpion charybdotoxin, a chimeric protein was constructed that reproduced the three-dimensional structure and partially reproduced the function of the parent beta hairpin, without perturbing the three dimensional structure of the scaffold [1]. Picosecond to hour time scale motions of charybdotoxin and the engineered protein were observed, in order to evaluate the dynamic consequences of the six deletions and eight mutations differentiating the two molecules. The chimeric protein dynamics were also compared to that of toxin alpha, in order to examine the beta hairpin motions in both structural contexts. Thus, 13C R1, R1rho and 1H-->13C nOe were measured for all the CalphaHalpha and threonine CbetaHbeta vectors. As the proteins were not labeled, accordion techniques combined to coherence selection by pulsed field gradients and preservation of magnetization following equivalent pathways were used to considerably reduce the spectrometer time needed. On one hand, we observed that the chimeric protein and charybdotoxin are subjected to similar picosecond to nanosecond time scale motions except around the modified beta sheet region. The chimeric protein also exhibits an additional millisecond time scale motion on its whole sequence, and its beta structure is less stable on a minute to hour time scale. On the other hand, when the beta hairpin dynamics is compared in two different structural contexts, i.e. in the chimeric protein and the curaremimetic toxin alpha, the picosecond to nanosecond time scale motions are fairly conserved. However, the microsecond to millisecond time scale motions are different on most of the beta hairpin sequence, and the beta sheet seems more stable in toxin alpha than in the chimera. The slower microsecond to hour time scale motions seem to be extremely sensitive to the structural context, and thus poorly transferred from one protein to another. PMID- 11054104 TI - Mitochondrial membrane potential differentiates cells resistant to apoptosis in hybridoma cultures. AB - Previous research has implicated mitochondrial physiology and, by extension, respiratory capacity in the initiation and progress of apoptosis of cells in culture and tissue environments. This hypothesis was tested by separating a hybridoma cell population into subpopulations of varying mitochondrial membrane potential (MMP) using Rhodamine 123 stain and fluorescence-activated cell sorter analysis and subjecting them to two apoptosis inducers, rotenone and staurosporin. Apoptotic death was characterized morphologically through the determination of apoptosis-related chromatin condensation and biochemically through the measurement of caspase-3 enzymatic activity. We found dramatic differences in the apoptotic death kinetics for the subpopulations, with the high MMP cells showing higher resistance to apoptotic death. After incubation with 30 microM rotenone, the low MMP cells exhibited one-third of the viability of the high MMP cells and a three-fold increase in the capsase-3 enzymatic activity. No changes were observed in the DNA content or the cell cycle distributions of the two cell subpopulations, which maintained their mean MMP difference after 20 generations. These results suggest that heterogeneity exists in mammalian cell populations with respect to mitochondrial physiology, which correlates with resistance to apoptotic death. PMID- 11054105 TI - Enzymes of hydrogen metabolism in Pyrococcus furiosus. AB - The genome of Pyrococcus furiosus contains the putative mbhABCDEFGHIJKLMN operon for a 14-subunit transmembrane complex associated with a Ni-Fe hydrogenase. Ten ORFs (mbhA-I and mbhM) encode hydrophobic, membrane-spanning subunits. Four ORFs (mbhJKL and mbhN) encode putative soluble proteins. Two of these correspond to the canonical small and large subunit of Ni-Fe hydrogenase, however, the small subunit can coordinate only a single iron-sulfur cluster, corresponding to the proximal [4Fe-4S] cubane. The structural genes for the small and the large subunits, mbhJ and mbhL, are separated in the genome by a third ORF, mbhK, encoding a protein of unknown function without Fe/S binding. The fourth ORF, mbhN, encodes a 2[4Fe-4S] protein. With P. furiosus soluble [4Fe-4S] ferredoxin as the electron donor the membranes produce H2, and this activity is retained in an extracted core complex of the mbh operon when solubilized and partially purified under mild conditions. The properties of this membrane-bound hydrogenase are unique. It is rather resistant to inhibition by carbon monoxide. It also exhibits an extremely high ratio of H2 evolution to H2 uptake activity compared with other hydrogenases. The activity is sensitive to inhibition by dicyclohexylcarbodiimide, an inhibitor of NADH dehydrogenase (complex I). EPR of the reduced core complex is characteristic for interacting iron-sulfur clusters with Em approximately -0.33 V. The genome contains a second putative operon, mbxABCDFGHH'MJKLN, for a multisubunit transmembrane complex with strong homology to the mbh operon, however, with a highly unusual putative binding motif for the Ni-Fe-cluster in the large hydrogenase subunit. Kinetic studies of membrane-bound hydrogenase, soluble hydrogenase and sulfide dehydrogenase activities allow the formulation of a comprehensive working hypothesis of H2 metabolism in P. furiosus in terms of three pools of reducing equivalents (ferredoxin, NADPH, H2) connected by devices for transduction, transfer, recovery and safety-valving of energy. PMID- 11054106 TI - Specificities of functionally expressed chalcone and acridone synthases from Ruta graveolens. AB - The common rue, Ruta graveolens L., expresses two types of closely related polyketide synthases that condense three malonyl-CoAs with N-methylanthraniloyl CoA or 4-coumaroyl-CoA to produce acridone alkaloids and flavonoid pigments, respectively. Two acridone synthase cDNAs (ACS1 and ACS2) have been cloned from Ruta cell cultures, and we report now the cloning of three chalcone synthase cDNAs (CHS1 to CHS3) from immature Ruta flowers. The coding regions of these three cDNAs differ only marginally, and the translated polypeptides show about 90% identity with the CHSs from Citrus sinensis but less than 75% with the Ruta endogeneous ACSs. CHS1 was functionally expressed in Eschericha coli and its substrate specificity compared with those of the recombinant ACS1 and ACS2. 4 Coumaroyl-CoA was the preferred starter substrate for CHS1, but cinnamoyl-CoA and caffeoyl-CoA were also turned over at significant rates. However, N methylanthraniloyl-CoA was not accepted. In contrast, highly active preparations of recombinant ACS1 or ACS2 showed low, albeit significant, CHS side activities with 4-coumaroyl-CoA, which on average reached 16% (ACS1) and 12% (ACS2) of the maximal activity determined with N-methylanthraniloyl-CoA as the starter substrate, while the conversion of cinnamoyl-CoA was negligible with both ACSs. The condensation mechanism of the acridone ring system differs from that of chalcone/flavanone formation. Nevertheless, our results suggest that very minor changes in the sequences of Ruta CHS genes are sufficient to also accommodate the formation of acridone alkaloids, which will be investigated further by site directed mutagenesis. PMID- 11054107 TI - Simple formal kinetics for the reversible uptake of molecular hydrogen by [Ni-Fe] hydrogenase from Desulfovibrio gigas. AB - Enzymatic electrocatalysis, triggered and monitored by means of cyclic voltammetry, enabled us to achieve quantitative analysis of the kinetics of the hydrogenase catalyzed process, in the 7.8-10.0 pH range, in the presence of an electrochemically generated redox mediator. The quantitative analysis can be carried out by use of a quite simple SRC model. The simplicity of the SRC model is compatible with the existence of multiple redox microstates, which can be combined in a potential adjustable triangular mechanism consisting of three catalytic cycles, which are formally identical from the kinetic point of view. The steps involved in the kinetic control of the reversible process are H2 uptake or production at the Ni-Fe catalytic site and the intermolecular electron transfer between the mediator and the distal [4Fe-4S] cluster. The related rate constants have been determined. For the two accompanying intramolecular electron transfers which proceed at equilibrium, the equilibrium constants were found to be in very good agreement with previously published data. PMID- 11054108 TI - Molecular cloning, functional complementation in Saccharomyces cerevisiae and enzymatic properties of phosphatidylinositol synthase from the protozoan parasite Toxoplasma gondii. AB - The obligate intracellular parasite Toxoplasma gondii, the causative agent of toxoplasmosis, switches between the rapidly dividing tachyzoite and the slowly replicating bradyzoite in intermediate hosts such as humans and domestic animals. We have recently identified a bradyzoite cDNA encoding a putative phosphatidylinositol (PtdIns) synthase using a subtractive library [Yahiaoui, B., Dzierszinski, F., Bernigaud, A., Slomianny, C., Camus, D., and Tomavo, S. (1999) Mol. Biochem. Parasitol. 99, 223-235]. Here, we report the cloning of another cDNA encoding PtdIns synthase that is exclusively expressed in the tachyzoite stage. The two transcripts are encoded by two different genes, which are stage specifically regulated. The deduced amino-acid sequence (258 amino acids with a calculated total molecular mass of 27.8 kDa) of the tachyzoite-specific cDNA shares a significant degree of identity (between 26.5 and 30.1%) to the PtdIns synthases from human, rat, Arabidopsis thaliana and yeast. Interestingly, the putative protein encompasses an N-terminal extension that is approximately 40 amino-acids longer than that of PtdIns synthases from other organisms. Functional complementation realized by tetrad analysis of segregants of a Saccharomyces cerevisiae PtdIns synthase-deficient mutant (PIS1/pis1:kanMX4) showed that only the T. gondii putative PtdIns synthase truncated at its N-terminal extension is able to restore the viability of the cells. We demonstrate that this protein expressed in yeast transformants is functionally active in the membrane preparation and requires manganese and magnesium ions for activity. To our knowledge, this is the first report on the molecular cloning and functional analysis of a gene encoding a PtdIns synthase in protozoan parasites. PMID- 11054109 TI - Effect of cadmium on the relationship between replicative and repair DNA synthesis in synchronized CHO cells. AB - Repair and replicative DNA synthesis were measured at different stages of the cell cycle in control and cadmium-treated Chinese hamster ovary (CHO-K1) cells. Cells were synchronized by counterflow centrifugal elutriation. Elutriation resulted in five repair and four replication subphases. On Cd treatment, repair synthesis was elevated in certain subphases. Replicative subphases were suppressed by Cd treatment, with some of the peaks almost invisible. The number of spontaneous strand breaks measured by random oligonucleotide primed synthesis assay showed a cell-cycle-dependent fluctuation in control cells and was greatly increased after Cd treatment throughout the S phase. Elevated levels of the oxidative DNA damage product, 8-oxodeoxyguanosine, were observed after Cd treatment, with the highest level in early S phase, which gradually declined as damaged cells progressed through the cell cycle. PMID- 11054110 TI - Interferon-gamma and lipopolysaccharide regulate the expression of Nramp2 and increase the uptake of iron from low relative molecular mass complexes by macrophages. AB - The natural resistance associated macrophage protein 2 (Nramp2) is a transporter that is involved in iron (Fe) uptake from transferrin (Tf) and low molecular mass Fe complexes. Here we describe the effect of the inflammatory mediators interferon-gamma (IFN-gamma) and lipopolysaccharide (LPS) on the expression of Nramp2 mRNA and Fe uptake by cells of the macrophage lineage. After incubation of the RAW264.7 macrophage cell line with LPS there was a sevenfold increase in the expression of the 2.3 kb Nramp2 mRNA transcript when compared with the control, but little effect on the Nramp2 3.1 kb transcript. These results indicate differential regulation of the two transcripts. Treatment with LPS resulted in an increase in 59Fe uptake from 59Fe-nitrilotriacetic acid, while transferrin receptor (TfR) mRNA levels and 59Fe uptake from 59Fe-Tf were decreased. Paradoxically, at the same time, an increase in iron regulatory protein (IRP)1 RNA-binding activity was observed. Incubation with IFN-gamma (50 U.mL-1) resulted in a marked decrease in TfR mRNA levels but had no effect on Nramp2 mRNA expression. Exposure of RAW264.7 cells to both IFN-gamma and LPS resulted in a fourfold increase in the Nramp2 2.3-kb transcript and a four to fivefold decrease in the 3.1-kb transcript when compared with the control. Furthermore, there was a decrease in TfR mRNA levels despite an increase in IRP1 RNA-binding activity and a marked increase in inducible nitric oxide synthase mRNA expression. Hence, TfR and Nramp2 mRNA expression did not appear to be regulated in a concerted manner. Similar responses to those found above for RAW264.7 cells were also observed in the J774 macrophage cell line and also for primary cultures of mouse peritoneal macrophages. These results are of interest as the TfR and Nramp2 are thought to act together during Fe uptake from Tf. This is the first report to demonstrate regulation of the Nramp2 mRNA transcripts by inflammatory mediators. PMID- 11054111 TI - Transcription inhibitors stimulate translation of 5' TOP mRNAs through activation of S6 kinase and the mTOR/FRAP signalling pathway. AB - We have analysed the effect of transcription inhibitors on the polysomal localization of 5' terminal oligopyrimidine (TOP-) mRNAs. It is known that, in vertebrates, the translation of this group of mRNAs is regulated according to the growth status of the cell. Mitogenic stimulation of quiescent cells induces a rapid recruitment of TOP mRNAs from translationally inactive light messenger ribonucleoprotein particles to polysomes. It was found that administration of transcription inhibitors to resting cells causes a similar collective translational activation of TOP mRNAs, without affecting global translation. A number of transcription inhibitors were tested in amphibian and mammalian cultured cells. Actinomycin D (act D), cordycepin, and 5, 6-dichloro-1-beta-D ribofuranosylbenzimidazole caused a similar activation whereas alpha-amanitin or low doses of act D did not induce the translational response. Concentrations of act D sufficient to induce TOP mRNA translation also induce 40S ribosomal protein S6 kinases 1 (S6K1) activation. Moreover at these concentrations of act D increased phosphorylation of 4E-BP1 was also observed, indicating the involvement of FRAP/mTOR. Consistent with this observation, pretreatment of resting cells with rapamycin suppresses the activation of TOP mRNA translation induced by act D. These results indicate that the effect of act D on translation is mediated by the S6Ks through FRAP/mTOR. PMID- 11054112 TI - Comparative functional characterization in vitro of heptosyltransferase I (WaaC) and II (WaaF) from Escherichia coli. AB - Heptosyltransferase II, encoded by the waaF gene of Escherichia coli, is a glycosyltransferase involved in the synthesis of the inner core region of lipopolysaccharide. The gene was subcloned from plasmid pWSB33 [Brabetz, W., Muller-Loennies, S., Holst, O. & Brade, H. (1997) Eur. J. Biochem. 247, 716-724] into a shuttle vector for the expression in the gram-positive host Corynebacterium glutamicum. The in vitro activity of the enzyme was investigated in comparison to that of heptosyltransferase I (WaaC) using as a source for the sugar nucleotide donor, ADP-LglyceroDmanno-heptose, a low molecular mass filtrate from a DeltawaaCF E. coli strain. Synthetic lipid A analogues varying in the acylation or phosphorylation pattern or both were tested as acceptors for the subsequent transfer of 3-deoxy-Dmanno-oct-2-ulosonic acid (Kdo) and heptose by successive action of Kdo transferase (WaaA), heptosyltransferase I (WaaC) and heptosyltransferase II (WaaF). The reaction products were characterized after separation by TLC and blotting with monoclonal antibodies specific for the acceptor, the intermediates and the final products. PMID- 11054113 TI - Conversion of the central [4Fe-4S] cluster into a [3Fe-4S] cluster leads to reduced hydrogen-uptake activity of the F420-reducing hydrogenase of Methanococcus voltae. AB - As in many other hydrogenases, the small subunit of the F420-reducing hydrogenase of Methanococcus voltae contains three iron-sulfur clusters. The arrangement of the three [4Fe-4S] clusters corresponds to the arrangement of [Fe-S] clusters in the [NiFeSe] hydrogenase of Desulfomicrobium baculatum. Many other hydrogenases contain two [4Fe-4S] clusters and one [3Fe-4S] cluster with a relatively high redox potential, which is located in the central position between a proximal and a distal [4Fe-4S] cluster. We have investigated the role of the central [4Fe-4S] cluster in M. voltae with regard to its effect on the enzyme activity and its spectroscopic properties. Using site-directed mutagenesis, we constructed a strain in which one cysteine ligand of the central [4Fe-4S] cluster was replaced by proline. The mutant protein was purified, and the [4Fe-4S] to [3Fe-4S] cluster conversion was confirmed by EPR spectroscopy. The conversion resulted in an increase in the redox potential of the [3Fe-4S] cluster by about 400 mV. The [NiFe] active site was not affected significantly by the mutation as assessed by the unchanged Ni EPR spectrum. The specific activity of the mutated enzyme did not show any significant differences with the artificial electron acceptor benzyl viologen, but its specific activity with the natural electron acceptor F420 decreased tenfold. PMID- 11054114 TI - A mutation affecting the association equilibrium of formyltransferase from the hyperthermophilic Methanopyrus kandleri and its influence on the enzyme's activity and thermostability. AB - Formyltransferase from Methanopyrus kandleri is composed of only one type of subunit of molecular mass 32 kDa. The enzyme is in a monomer/dimer/tetramer association equilibrium, the association constant being affected by lyotropic salts. Oligomerization is required for enzyme activity and thermostability. We report here on a subunit interface mutation (R261E) which affects the dimer/tetramer part of the association equilibrium of formyltransferase. With the mutant protein it was shown that tetramerization is not required for activity but is necessary for high thermostability. PMID- 11054115 TI - Redox potentials and their pH dependence of D-amino-acid oxidase of Rhodotorula gracilis and Trigonopsis variabilis. AB - The redox potentials and pH characteristics of D-amino-acid oxidase (EC 1.4.3.3; DAAO) from the yeast Rhodotorula gracilis and Trigonopsis variabilis were measured in the pH range 6.5-8.5 at 15 degrees C. In the free enzyme form, the anionic red semiquinone is quantitatively formed in both DAAOs, indicating that a two single-electron transfer mechanism is active. The semiquinone species is also thermodynamically stable, as indicated by the large separation of the single electron transfer potentials. The first electron potential is pH-independent, while the second electron transfer is pH-dependent exhibiting a approximately -60 mV/pH unit slope, consistent with a one-electron/one-proton transfer. In the presence of the substrate analogue benzoate, the two-electron transfer is the thermodynamically favoured process for both DAAOs, with only a quantitative difference in the stabilization of the anionic semiquinone. Clearly binding of the substrate (or substrate analogue) modulates the redox properties of the two enzymes. In both cases, in the presence and absence of benzoate, the slope of Em vs. pH (-30 mV/pH unit) corresponds to an overall two-electron/one-proton transfer in the reduction to yield the anionic reduced flavin. This behaviour is similar to that reported for DAAO from pig kidney. The differences in potentials and the stability of the semiquinone intermediate measured for the three DAAOs probably stem from different isoalloxazine environments. In the case of R. gracilis DAAO, the low stability of the semiquinone form in the DAAO-benzoate complex can be explained by the shift in position of the side chain of Arg285 following substrate analogue binding. PMID- 11054116 TI - A novel type of arabinoxylan arabinofuranohydrolase isolated from germinated barley analysis of substrate preference and specificity by nano-probe NMR. AB - An arabinoxylan arabinofuranohydrolase was isolated from barley malt. The enzyme preparation, Ara 1, contained two polypeptides with apparent molecular masses of approximately 60 and approximately 66 kDa, a pI of 4.55 and almost identical N terminal amino-acid sequences. With p-nitrophenyl alpha-L-arabinofuranoside (pNPA) as substrate, Ara 1 exhibited a Km of 0.5 mM and a Vmax of 6.7 micromol. min-1.(mg of protein)-1. Maximum activity was displayed at pH 4.2 and 60 degrees C, and, under these conditions, the half-life of the enzyme was 8 min. The Ara 1 preparation showed no activity against p-nitrophenyl alpha-L-arabinopyranoside or p-nitrophenyl beta-D-xylopyranoside. Substrate preference and specificity were investigated using pure oligosaccharides and analysis by TLC and nano-probe NMR. Ara 1 released arabinose from high-molecular-mass arabinoxylan and arabinoxylan derived oligosaccharides but was inactive against linear or branched-chain arabinan. Arabinose was readily released from both singly and doubly substituted xylo-oligosaccharides. Whereas single 2-O-linked and 3-O-linked arabinose substituents on non-reducing terminal xylose were released at similar rates, there was a clear preference for 2-O-linked arabinose on internal xylose residues. When Ara 1 acted on oligosaccharides with doubly substituted, non reducing terminal xylose, the 3-O-linked arabinose group was preferred as the initial point of attack. Oligosaccharides with doubly substituted internal xylose were poor substrates and no preference could be determined. The enzyme described here is the first reported arabinoxylan arabinofuranohydrolase which is able to release arabinose from both singly and doubly substituted xylose, and it hydrolyses p-nitrophenyl alpha-L-arabinofuranoside at a rate similar to that observed for oligosaccharide substrates. PMID- 11054117 TI - Peroxovanadate induces tyrosine phosphorylation of phosphoinositide-dependent protein kinase-1 potential involvement of src kinase. AB - Phosphoinositide-dependent protein kinase-1 (PDK1) is a recently identified kinase that phosphorylates and activates protein kinase B (PKB). Activation of PKB by insulin is linked to its translocation from the cytosol to the plasma membrane. However, no data are available yet concerning the localization of PDK1 in insulin-sensitive tissue. Using isolated adipocytes, we studied the effect of insulin and of an insulin-mimicking agent peroxovanadate on the subcellular localization of PDK1. In unstimulated adipocytes, overexpressed PDK1 was mostly cytosolic with a low amount associated to membranes. Peroxovanadate stimulation induced the redistribution of PDK1 to the membranes while insulin was without effect. This peroxovanadate effect was dependent on phosphatidylinositol 3,4,5 triphosphate [PtdIns(3,4,5)P3] production as inhibition of PtdIns 3-kinase by wortmannin or deletion of the PH domain of PDK1 prevented the peroxovanadate induced translocation of PDK1. Further, peroxovanadate-treatment induced a tyrosine phosphorylation of PDK1 which was wortmannin insensitive and did not require the PH domain of PDK1. An inhibitor of Src kinase (PP2) decreased the peroxovanadate-induced PDK1 tyrosine phosphorylation and overexpression of v-Src stimulated this phosphorylation. Mutation of tyrosine 373 of PDK1 abolished the v Src induced PDK1 tyrosine phosphorylation and partially reduced the effect of peroxovanadate. Our findings suggest that PDK1 could be a substrate for tyrosine kinases and identify Src kinase as one of the tyrosine kinases able to phosphorylate PDK1. PMID- 11054118 TI - Effects of proline mutations in the major house dust mite allergen Der f 2 on IgE binding and histamine-releasing activity. AB - Der f 2 is the major group 2 allergen from house dust mite Dermatophagoides farinae and is composed of 129 amino-acid residues. Wild-type and six proline mutants of Der f 2 (P26A, P34A, P66A, P79A, P95A, and P99A) expressed in Escherichia coli were refolded and purified. Formations of intramolecular disulfide bonds in the purified proteins were confirmed correct. The apparent molecular masses analyzed by gel-filtration were 14-15 kDa. The IgE-binding capacity in the sera of seven mite-allergic patients, inhibitory activity for IgE binding to immobilized wild-type Der f 2, and activity to stimulate peripheral blood basophils to release histamine in two volunteers were analyzed. P95A and P99A, which slightly differed from the wild-type Der f 2 in their CD spectrum, showed reduced IgE-binding, reduced inhibitory activity, and less histamine releasing activity than the wild-type. P34A also showed reduced allergenicity. Considering that Pro95, Pro99 and Pro34 are closely located in loops at one end of the tertiary structure of Der f 2, we concluded that these loop regions included an IgE-binding site common to all tested patients. P66A showed reduced IgE-binding in two sera out of seven. P26A and P79A showed no reduced allergenicity. However, in immunoblot analysis after SDS/PAGE under reduced conditions, P79A showed no or markedly reduced IgE-binding while the other mutants showed IgE-binding corresponding to that in the assay using correctly refolded proteins. This suggests that Pro79 is involved in refolding of Der f 2. The findings in this study are important for the understanding of the antigenic structure of mite group 2 allergens and for manipulation of the allergens for specific immunotherapy. PMID- 11054119 TI - Characterization of the NAD+ binding site of Candida boidinii formate dehydrogenase by affinity labelling and site-directed mutagenesis. AB - The 2',3'-dialdehyde derivative of ADP (oADP) has been shown to be an affinity label for the NAD+ binding site of recombinant Candida boidinii formate dehydrogenase (FDH). Inactivation of FDH by oADP at pH 7.6 followed biphasic pseudo first-order saturation kinetics. The rate of inactivation exhibited a nonlinear dependence on the concentration of oADP, which can be described by reversible binding of reagent to the enzyme (Kd = 0.46 mM for the fast phase, 0.45 mM for the slow phase) prior to the irreversible reaction, with maximum rate constants of 0.012 and 0.007 min-1 for the fast and slow phases, respectively. Inactivation of formate dehydrogenase by oADP resulted in the formation of an enzyme-oADP product, a process that was reversed after dialysis or after treatment with 2-mercaptoethanol (> 90% reactivation). The reactivation of the enzyme by 2-mercaptoethanol was prevented if the enzyme-oADP complex was previously reduced by NaBH4, suggesting that the reaction product was a stable Schiff's base. Protection from inactivation was afforded by nucleotides (NAD+, NADH and ADP) demonstrating the specificity of the reaction. When the enzyme was completely inactivated, approximately 1 mol of [14C]oADP per mol of subunit was incorporated. Cleavage of [14C]oADP-modified enzyme with trypsin and subsequent separation of peptides by RP-HPLC gave only one radioactive peak. Amino-acid sequencing of the radioactive tryptic peptide revealed the target site of oADP reaction to be Lys360. These results indicate that oADP inactivates FDH by specific reaction at the nucleotide binding site, with negative cooperativity between subunits accounting for the appearance of two phases of inactivation. Molecular modelling studies were used to create a model of C. boidinii FDH, based on the known structure of the Pseudomonas enzyme, using the MODELLER 4 program. The model confirmed that Lys360 is positioned at the NAD+-binding site. Site directed mutagenesis was used in dissecting the structure and functional role of Lys360. The mutant Lys360-->Ala enzyme exhibited unchanged kcat and Km values for formate but showed reduced affinity for NAD+. The molecular model was used to help interpret these biochemical data concerning the Lys360-->Ala enzyme. The data are discussed in terms of engineering coenzyme specificity. PMID- 11054120 TI - Conformations of the regulatory domain of cardiac troponin C examined by residual dipolar couplings. AB - Conformations of the regulatory domain of cardiac troponin C (cNTnC) were studied by means of residual dipolar couplings measured from samples dissolved in dilute liquid crystals. Changes in the main chain HN residual dipolar couplings revealed a conformational change in cNTnC due to the complexation with the second binding region (amino acids 148-163) of cardiac troponin I (cTnI). Formation of the complex is accompanied with a molecular realignment in the liquid crystal. The residual dipolar couplings measured for apo-cNTnC and the complex with TnI were in agreement with the values computed from the corresponding closed and open solution structures, whereas for the calcium-loaded conformation the correlation and quality factor were only modest. Ca2+-cNTnC may be subject to conformational exchange. The data support the model that cardiac troponin C functions as a calcium-dependent open-closed switch, such as the skeletal troponin C. PMID- 11054121 TI - Heme-(hydro)peroxide mediated O- and N-dealkylation. A study with microperoxidase. AB - The mechanism of microperoxidase-8 (MP-8) mediated O- and N-dealkylation was investigated. In the absence of ascorbate (peroxidase mode), many unidentified polymeric products are formed and the extent of substrate degradation correlates (r = 0.94) with the calculated substrate ionization potential, reflecting the formation of radical intermediates. In the presence of ascorbate (P450 mode) formation of polymeric products is largely prevented but, surprisingly, dealkylation is not affected. In addition, aromatic hydroxylation and oxidative dehalogenation is observed. The results exclude a radical mechanism and indicate the involvement of a (hydro)peroxo-iron heme intermediate in P450-type of heteroatom dealkylation. PMID- 11054122 TI - Protein disulfide isomerase-mediated cell-free assembly of recombinant interleukin-12 p40 homodimers. AB - Interleukin-12 (IL-12) is a heterodimeric cytokine composed of two subunits, p35 and p40. The disulfide-linked homodimer (p40)2 has been shown to be a potent IL 12 antagonist. In the present study, the p40 subunit was refolded from Escherichia coli inclusion bodies. Formation of (p40)2 was greatly increased in a redox buffer containing reduced and oxidized glutathione, but was not significantly affected by the cosolvents urea, GdnHCl or Chaps. While protein disulfide isomerase (PDI), GroEL/ES or DnaK/J/GrpE suppressed aggregation during refolding of p40, only DnaK/J/GrpE and PDI enhanced p40 dimerization. Oxidative assembly of p40 into (p40)2 by PDI, but not suppression of aggregation, was strongly dependent on inclusion of BSA in the refolding buffer. It is concluded that both chaperone-like and disulfide isomerase effects are essential for correct folding of p40 into dimers. PMID- 11054123 TI - Phospholipases A2 from Callosellasma rhodostoma venom gland cloning and sequencing of 10 of the cDNAs, three-dimensional modelling and chemical modification of the major isozyme. AB - Callosellasma rhodostoma (Malayan pitviper) is a monotypic Asian pitviper of medical importance. Three acidic phospholipases A2 (PLA2s) and one basic PLA2 homolog were purified from its venom while 10 cDNAs encoding distinct PLA2s were cloned from venom glands of a Thailand specimen of this species. Complete amino acid sequences of the purified PLA2s were successfully deduced from their cDNA sequences. Among the six un-translated PLA2 cDNAs, two apparently result from recombination of its Lys49-PLA2 gene with its Asp49-PLA2 genes. The acidic PLA2s inhibit platelet-aggregation, while the noncatalytic PLA2-homolog induces local edema. This basic PLA2-homolog contains both Asp49 and other, unusual substitutions unique for the venom Lys49-PLA2 subtype (e.g. Leu5, Trp6, Asn28 and Arg34). Three-dimensional modelling of the basic protein revealed a heparin binding region, and an abnormal calcium-binding pocket, which may explain its low catalytic activity. Oxidation of up to six of its Met residues or coinjection with heparin reduced its edema-inducing activity but methylation of its active site His48 did not. The distinct Arg/Lys-rich and Met-rich region at positions 10 36 of the PLA2 homolog presumably are involved in its heparin-binding and the cell membrane-interference leading to edema and myotoxicity. PMID- 11054124 TI - Examining the zinc binding site of the amyloid-beta peptide. AB - The amyloid beta-peptide (Abeta) is a principal component of insoluble amyloid plaques which are characteristic neuropathological features of Alzheimer's disease. Abeta also exists as a normal soluble protein that undergoes a pathogenic transition to an aggregated, fibrous form. This transition can be affected by extraneous proteinaceous and nonproteinaceous elements, such as zinc ions, which may promote aggregation and/or stabilization of the fibrils. Protein chelation of zinc is typically mediated by histidines, cysteines and carboxylates. Previous studies have demonstrated that the Abeta-Zn2+ binding site is localized within residues 6-28 and that histidines may serve as the principal sites of interaction. To localize key residues within this region, a series of Abeta peptides (residues 1-28) were synthesized that contained systematic His/Ala substitutions. Circular dichroism and electron microscopy were used to monitor the effects of Zn2+ on the peptide beta-sheet conformation and fibril aggregation. Our results indicate that substitution of either His13 or His14 but not His6 eliminates the zinc-mediated effects. These observations indicate a specific zinc binding site within Abeta that involves these central histidine residues. PMID- 11054125 TI - The expression of Cox17p in rodent tissues and cells. AB - Previous works have reported the isolation of a novel polypeptide from porcine heart. Structural analysis has shown that it is a mammalian homologue of Cox17p, believed essential for the assembly of functional cytochrome c oxidase and delivery of copper ions to the mitochondrion for insertion into the enzyme in yeast. Although the human, mouse and porcine homologs of this small protein have already been cloned or purified, the function of Cox17p in the mammalian system has not yet been elucidated. To investigate the physiological function of Cox17p in mammals, we performed Northern blot analysis using probes containing the mouse and rat sequences obtained by RT-PCR. The hybridization signals were detected in all mouse tissues, but notably intense signals were observed in heart, brain and kidney RNA samples. Some of the neuroendocrine and endocrine cell lines showed higher expression levels than fibroblasts. The highest expression level of Cox17p mRNA in mouse brain was observed in the pituitary sample. While in rat heart, Cox17p mRNA expression was detected from early development, in rat brain, embryonic and postnatal changes in the expression were observed. Immunocytochemical analysis showed that Cox17p immunoreactivity was strong in the pituitary cell line, AtT-20. These findings suggested that Cox17p is not only part of the respiratory chain but also involved in brain and endocrine functions. PMID- 11054126 TI - Characterization of the general odorant-binding protein 2 in the molecular coding of odorants in Mamestra brassicae. AB - The general odorant-binding protein 2 of Mamestra brassicae males has been purified from antennal extracts and examined in binding assays with pheromone components of this species and a behavioral antagonist, cis-11-hexadecenol. The protein showed high affinity for the latter compound and no affinity for the pheromone components. In addition, expression of the protein, studied by in situ hybridization, was restricted to the long sensilla trichodea, which house the neuron that responds to cis-11-hexadecenol. The expression in a functionally defined population of sensilla, together with binding specificity and previous electrophysiological data, suggest an unsuspected role for the general odorant binding protein 2 in M. brassicae. It may be involved in the transduction process for the behavioral antagonist to which neurons are specifically tuned and always cocompartmentalized in long trichodeal hairs, with neurons responding to the major pheromonal compound, cis-11-hexadecenyl acetate. These data are consistent with the involvement of odorant-binding proteins in the fine discrimination between pheromone and antagonist, which is related to avoidance of interspecific mating mistakes. PMID- 11054127 TI - Heart and carotid artery disease in stroke patients with intermittent claudication. AB - Much has been published on the natural history of intermittent claudication (IC), but little is known about the clinical features of stroke patients with IC. The purpose of this study was to examine clinical features and risk factors in stroke patients with or without IC, including heart disease and carotid artery disease. A hospital-based study was conducted of 3901 stroke patients, who were prospectively coded and entered into a computerized databank. Of these patients, 219 had symptoms of IC. Patients were subdivided by age into 10-year categories. There were at least 12 times more non-IC than IC patients in each category. An age-matched random sample was obtained containing 12 non-IC cases for each IC case, resulting in 219 cases of IC and 2628 non-IC cases. The prevalence of IC in the total stroke population was 5.6%. IC prevalence increased sharply with age until about 70 years. Cardiac ischaemia and internal carotid artery (ICA) disease were significantly more frequent in stroke with IC than without IC. IC patients also exhibited a higher prevalence of atherosclerotic disease as well as other risk factors such as smoking, hypercholesterolaemia, elevated haematocrit, and family history of stroke. Ischaemic heart disease and ICA disease are especially common in stroke with IC. IC, large artery disease and stroke share similar risk factors. IC symptoms in stroke patients may indicate extensive generalized atherosclerosis. PMID- 11054128 TI - Juvenile parkinsonism: a term in search of an identity. PMID- 11054129 TI - Juvenile parkinsonism: a heterogeneous entity. AB - We studied the clinical features, laboratory investigation, management and natural history of a cohort of patients with Juvenile Parkinsonism (JP), seen at a tertiary referral centre. JP was defined as Parkinsonism with onset at age 20 years or less. Six patients (five male, one female) entered the study. The mean age at onset of Parkinsonism was 12.5 years (range 7-19) and the mean follow-up time was 49.3 months (range 40-57). Bradykinesia, rigidity, and postural instability were observed in all patients and five subjects had tremor. Dystonia was present in four subjects. Other clinical features were dementia (five subjects), supranuclear ophthalmoparesis (five subjects), seizures (three subjects), multifocal myoclonus (one subject), decreased deep reflexes (one subject), pyramidal signs (one subject). Family history of Parkinson's disease (PD) was positive in one subject. Work-up for Wilson's disease was negative in all patients. Neuroimaging studies showed cortical atrophy in two subjects and mild brainstem atrophy in two others. Sea-blue histiocytes were found in one subject. L-dopa improved the Parkinsonism in all subjects but four rapidly developed fluctuations and dyskinesias, requiring, in one, stereotaxic surgery. After a mean disease duration of 6.5 years, five subjects require assistance for performance of all daily activities. JP is a heterogeneous clinical entity. In the majority of patients, no underlying cause is identified. The unusual clinical features suggest most subjects have a CNS degenerative disease distinct from PD. There is, however, evidence suggesting that PD may rarely cause JP. Gangliosidosis is another cause of L-dopa-responsive JP. Regardless of the cause, in the present study JP displays an aggressive and rapidly progressive course in most patients. PMID- 11054130 TI - Cognitive processes involved in delayed non-matching-to-sample performance in Parkinson's disease. AB - Visual recognition memory was assessed in terms of delay duration, memory load and amount of interference(s) in non-demented patients with Parkinson's disease (PD) using an automated delayed non-matching-to-sample (DNMS) task with trial unique stimuli. Special attention was focused on the different cognitive functions engaged by these patients in solving this recognition memory task. Thirteen patients with PD, carefully selected according to their stable regimen and anticholinergic medication, were compared to 12 controls matched by age and educational level. Besides the DNMS task, a neuropsychological battery that included tasks carefully selected according to processes potentially required to perform the DNMS task (e.g. attention, executive functions, visual discrimination and motor speed) was administered to the subjects. As compared with controls, patients with PD showed a deficit on most DNMS subscores, except those requiring the least cognitive load. The correlative analysis between the DNMS and other neuropsychological tasks suggests involvement of long-term memory mainly in the DNMS performance for the control group, contrasting with a major involvement of executive functions for the patients with PD. These data indicate that visual recognition memory impairment in non-demented patients with PD is largely due to an executive dysfunction, notably in working memory. Several hypotheses are proposed concerning the neuronal substrates underlying the impairment on the visual DNMS task in PD. PMID- 11054131 TI - Epidemiology of dementia in Nigeria: results from the Indianapolis-Ibadan study. AB - We determined the prevalence of dementia in a cohort of 2494 elderly Nigerians residents in Idikan Community, Ibadan, Nigeria. We utilized the Community Screening Instrument for Dementia to select subjects for clinical assessment in the second stage. The findings were compared with those of 2212 African Americans living in Indianapolis, studied simultaneously using similar methodology. The overall age-adjusted prevalence rates of dementia and Alzheimer's disease in Ibadan were 2.29% and 1.41%, respectively. These rates were much lower than the respective values of 8.24% and 6.24% obtained for African Americans. In Ibadan, Alzheimer's disease accounted for 64.3% of the cases, with old age and female gender being the significant risk factors, whilst 'living with others' appeared to be protective (P < 0.05). Amongst African Americans, on the other hand, old age, rural living below the age of 19 years, low educational attainment and family history of dementia were the risk factors. There was a lack of association between Alzheimer's disease and possession of the apolipoprotein E epsilon4 allele in the Nigerian sample, unlike the finding in African Americans, where significant association was shown. In addition, the frequencies of the vascular risk factors investigated were lower in Nigerians. Our results showed lower prevalence of dementia and Alzheimer's disease in Nigerians, compared with African Americans. There was no association between Alzheimer's disease and apolipoprotein E (epsilon4 allele) in the former. The differences in the frequencies of vascular risk factors between Nigerian subjects and African Americans would suggest involvement of environmental factors in disease process. PMID- 11054132 TI - The level of serum lipids, vitamin E and low density lipoprotein oxidation in Wilson's disease patients. AB - The aim of this study was to estimate the level of lipids and of the main serum antioxidant, alpha-tocopherol (vitamin E), and to evaluate the susceptibility of low density lipoprotein (LDL) to oxidation in Wilson's disease patients. It was assumed that enhanced LDL peroxidation caused by high copper levels could contribute to the injury of liver and other tissues. The group investigated comprised 45 individuals with Wilson's disease treated with penicillamine or zinc salts and a control group of 36 healthy individuals. Lipids were determined by enzymatic methods, alpha-tocopherol by high performance liquid chromatography, the susceptibility of LDL to oxidation in vitro by absorption changes at 234 nm during 5 h and end-products of LDL lipid oxidation as thiobarbituric acid reacting substances. In Wilson's disease patients total cholesterol, LDL cholesterol and alpha-tocopherol levels were significantly lower compared with the control group. No difference in LDL oxidation in vitro between the patients and the controls was stated. CONCLUSION: enhanced susceptibility of isolated LDL for lipid peroxidation in vitro was not observed in Wilson's disease patients. One cannot exclude, however, that because of low alpha-tocopherol level lipid peroxidation in the tissues can play a role in the pathogenesis of tissue injury in this disease. PMID- 11054133 TI - Frequency of His1069Gln and Gly1267Lys mutations in Polish Wilson's disease population. AB - Wilson's disease is an autosomal recessive disorder. More than 60 mutations of the Wilson's disease gene have been described so far. We have analysed 148 Polish Wilson's disease patients from 95 families for His1069Gln and Gly1267Lys mutations and correlated this finding with age and clinical form of the disease at presentation. To identify these mutations, single strand conformation polymorphism analysis was performed. In our group there were 94 patients with neurological presentation, 28 with hepatic presentation, whilst 26 were in a pre clinical stage of the disease. His1069Gln mutation was present on 171 (57%) of the 296 studied chromosomes, and Gly1267Lys mutation was present on 27 chromosomes (9.1%). Most of our patients were homozygous or heterozygous for His1069Gln mutation (39.9% and 30.4%, respectively); 4% of the patients were homozygous for Gly1267Lys mutation and 5.4% had both of these described mutations on their chromosomes. His1069Gln and Gly1267Lys mutations occurred often in our Wilson's disease patient population but we did not find any relationship between investigated mutations and the clinical form of Wilson's disease or age of first symptoms. PMID- 11054134 TI - Patterns of cerebrovascular symptomatology associated with carotid atheroma. AB - The aim of this study was to identify ultrasonic tissue characteristics and stenosis of carotid plaques that correspond to amaurosis fugax, hemispheric transient ischaemic attack, and stroke. At total of 146 symptomatic carotid plaques (136 patients) associated with amaurosis fugax, hemispheric transient ischaemic attack, stroke, and having 50-99% stenosis on duplex, were studied. These plaques were imaged on duplex, captured in a computer and their grey scale median was evaluated to distinguish the dark (low grey scale median) from the bright (high grey scale median) plaques. Stenosis was assessed on duplex. The amaurosis fugax group corresponded to carotid plaques with low grey scale median and severe stenosis, as contrasted with the other two groups (hemispheric transient ischaemic attack and stroke) (P < 0.05). These results suggested that amaurosis fugax was dependent only on the instability of carotid plaques, whereas hemispheric transient ischaemic attack and stroke were both dependent on carotid plaques and other pathogenetic factors. PMID- 11054135 TI - Assessment of corticodiaphragmatic pathway and pulmonary function in acute ischemic stroke patients. AB - The aims of this study were to investigate the effect of stroke on the corticodiaphragmatic pathway and to clarify the relationships between neurophysiological data and degree of motor disability, site of infarction in CT scan, diaphragmatic excursion, blood gases and pulmonary function in stroke patients. The corticodiaphragmatic pathway was assessed using magnetic stimulation of the scalp sites and cervical roots. The study included 34 sequentially selected patients out of 250 patients with acute ischemic stroke. Twenty-five (age and sex matched) volunteers served as controls. Sixteen patients had cortical infarction, thirteen had subcortical infarction and five had both cortical and subcortical infarction. The mean Scandinavian Stroke Scale was 32.2. Decreased diaphragmatic excursion was observed in 41% of the patients. Twenty four patients (70.5%) had abnormal magnetic evoked potentials (MEPs) of the affected hemisphere. In five patients MEPs were unelicitable from the affected hemisphere. The remaining nineteen patients had abnormal values of both cortical latency and central conduction time (CCT). Cortical latency, CCT, amplitude of compound muscle action potentials (CMAPs) and excitability threshold of the affected hemisphere were significantly altered compared to both the unaffected hemisphere and the control group. The patients with hemiplegia had a greater degree of hypoxia, hypocapnia and decreased serum bicarbonate level compared to the control group. Additionally, hemiplegic patients had a different degree of respiratory dysfunction. A statistically significant association was found between neurophysiological data and disability score, diaphragmatic excursion, site of infarction in CT scan and degree of respiratory dysfunction. Central diaphragmatic impairment may occur in acute stroke and could contribute to the occurrence of hypoxia in those patients. PMID- 11054136 TI - Delayed visual P3 in unilateral thalamic stroke. AB - The P3 potential is accepted as a neurophysiological correlate of memory and attention. Delayed latencies were reported in different forms of dementias. Although the generator sites are still under debate, the thalamus may play a crucial role. The aim of this study was to investigate the influence of an unilateral thalamic ischaemic infarction on P3 generation. The event-related P3 component of six patients (2 male, four female; mean age 47 years, range 22-63 years) with unilateral thalamic ischaemic infarction was studied and compared to age-matched controls (five male, nine female; mean age 45.8 years; range 22-69 years). All patients underwent full clinical examination, CCT, and MRI scan. P3 potentials were recorded with a visual three stimulus discrimination paradigm. The mean P3 latency of the patient group to the target stimulus was delayed (469.7 ms, SD = 36.8) compared with the controls (378.8 ms, SD = 51.5; P < 0. 05). The mean P3 latency to the unexpected stimulus was delayed in patients with thalamic infarction compared with controls [477 ms (SD = 46.6) vs. 381.2 ms (SD = 48.5); P < 0.001). Delayed P3 components of the event-related potential (ERP) were recorded in six patients with unilateral thalamic infarction, suggesting an important role of the thalamus in the generation of the P3 potential. PMID- 11054137 TI - Evaluation of a German version of the Rivermead Mobility Index (RMI) in acute and chronic stroke patients. AB - The English Rivermead Mobility Index (RMI) has been proposed as a simple, valid and reliable measure in stroke rehabilitation. A German version was established and validated in two centres. In centre A 46 acute (median: 3.0 days after onset) and in centre B 151 chronic (median: 88.0 days after onset) stroke patients participated. Interrater reliability of the German RMI was tested in 12 subjects in the acute stage of stroke and was found to be statistically significant (r = 0.98, P < 0.0001). In centre A, a statistically significant correlation was found between the German RMI and the 10-m walk time at baseline (r = 0.73, P < 0.0001) and after three weeks (r = 0.92, P < 0.0001). In centre B, the German RMI correlated significantly with the motor part of the Functional Independence Measure (motor-FIM) on admission (r = 0.78, P < 0.0001) and after three weeks (r = 0.79, P < 0.0001), respectively. The change of the RMI correlated significantly with the change in 10-m walk time in acute patients (r = 0.87, P < 0.0001) and with the change in motor-FIM in chronic patients (r = 0.54, P < 0.0001). A moderate ceiling-effect was detected in the chronic study population. The German RMI appears to be a reliable, valid and responsive measure for mobility disability in acute and chronic stroke patients. PMID- 11054138 TI - Abnormal perception of the tonic vibration reflex in idiopathic focal dystonia. AB - Although the pathophysiological basis of idiopathic focal dystonia (IFD) remains unclear, we recently reported abnormal perception of the tonic vibration reflex (TVR) in the biceps brachii in IFD. In this study we examined whether the abnormality affects muscles other than the biceps brachii. A 100-Hz vibration stimulating predominantly the muscle spindle afferent was transcutaneously applied to one muscle tendon of the triceps brachii, the wrist extensor and flexor muscles in 29 subjects with IFD (18 with torticollis, 9 with writer's cramp, 2 with blepharospasm) and 15 control subjects. The blindfolded subjects were instructed to copy any perceived movement with the opposite tracking arm. The elbow or wrist angle changes were quantified electronically. The TVR and subjects' perception of the TVR were evaluated by angle movements of the vibrated joint and of the tracking joint, respectively. Perception of the TVR was significantly reduced in dystonic subjects at both elbow and wrist joints, while magnitude of the TVR did not differ between the two groups. Abnormal central perception of the TVR is a feature of IFD, suggesting a widespread involvement of abnormal muscle spindle afferent processing in IFD. PMID- 11054139 TI - Different phenotypes of Friedreich's ataxia within one 'pseudo-dominant' genealogy: relationships between trinucleotide (GAA) repeat lengths and clinical features. AB - We examined a large Turkmen family with 'pseudo-dominant' inheritance of Friedreich's ataxia resulting from consanguineous marriage of a Friedreich's ataxia patient to a heterozygote carrying an ancestral mutated allele. Two distinct phenotypes of the disease co-segregated within this genealogy. Two brothers from the younger generation exhibited 'classical' Friedreich's ataxia with onset of symptoms before 10 years and a rapidly progressive course. In contrast, three patients (two sisters from the younger generation and their father) had a more benign phenotype of late-onset Friedreich's ataxia with the onset at 26, 45 and 48 years and slow progression over decades. The patients with 'classical' Friedreich's ataxia were homozygous for a common ancestral expanded allele of the X25 gene containing 700-800 GAA repeats, while the patients with late-onset Friedreich's ataxia had two different mutated alleles, the shorter 250 repeat expansion of paternal origin and the longer 700-repeat expansion of maternal origin. One may conclude that clinical variability of Friedreich's ataxia in our patients is accounted for predominantly by a modifying effect of one of the two (shorter or longer) expanded alleles inherited from their affected father. Our observation clearly demonstrates the significance of variable-sized alleles for the phenotypic expression of the disease. PMID- 11054140 TI - Elevation of serum soluble E-selectin and antisulfoglucuronyl paragloboside antibodies in amyotrophic lateral sclerosis. AB - Immunological abnormality is often found in amyotrophic lateral sclerosis (ALS). Antibodies to sulfoglucuronyl paragloboside (SGPG) were reported in ALS, although the pathogenetic significance of the antibodies is still unknown. We have already demonstrated that SGPG, a unique glycolipid, is present in both peripheral nerve and vascular endothelial cells. To investigate whether serum anti-SGPG antibodies would participate in activation and/or injury of endothelial cells in ALS, we examined serum anti-SGPG antibodies in association with serum soluble E- and P selectins, which are markers of activated endothelial cells, in 25 patients with ALS and 14 age-matched patients with other neurological diseases (ONDs) using the microtiter-ELISA method. Seven out of 25 ALS patients had anti-SGPG antibodies. Levels of sE-selectin were significantly higher in patients with ALS (48.5 +/- 23.4 ng/ml) compared with ONDs (24.0 +/- 11.8 ng/ml) (P < 0.005). Four out of seven ALS patients with anti-SGPG antibodies had concomitantly high sE-selectin levels. The mean sE-selectin levels were higher in patients with anti-SGPG antibodies (61.9 +/- 25.2 ng/ml) than in those without anti-SGPG antibodies (43.3 +/- 21.1 ng/ml). Anti-SGPG antibodies may take part in the activation and/or injury of endothelial cells. The increased expression of E-selectin may be related to an immunological process in some ALS patients. PMID- 11054141 TI - Extensive radiculopathy: a manifestation of intracranial hypertension. AB - We report two patients with severe radiculopathy due to elevated intracranial pressure (ICP) resulting from idiopathic intracranial hypertension (IHH) in one, and cerebral venous sinus thrombosis (CVT) in the other. Our aim is to document this unique association, which escaped diagnosis in both patients. PMID- 11054142 TI - Body packer: cocaine intoxication, causing death, masked by concomitant administration of major tranquilizers. AB - Cocaine, derived from the leaves of the shrub Erythroxylon coca, which grows on the slopes of the Andes, remains one of the most widely abused illicit drugs (Johnson et al., 1993). Its abuse appears to be increasing and as a result, so is its trafficking across borders, with ever-increasing sophistication of concealment (Rouse, 1992). Over the past few years, cases of cocaine intoxication have been reported, resulting from ruptured packets of cocaine that have been swallowed, or inserted into the vagina or rectum by couriers (drug smugglers), so called 'body packers' or 'mules' (Westli and Mittleman, 1981; Ricaurte and Langston, 1995). Cocaine is a powerful sympathomimetic and central nervous system stimulant, an overdose of which causes primarily cardiac, neurological and psychiatric effects (Ricaurte and Langston, 1995). Acute toxicity is dose-related and is characterized in the first place by its sympathomimetic effects, which include tachycardia, hypertension and hyperthermia arrythmias, followed by seizures. Brainstem depression and cardio-respiratory collapse, stroke, coma, intracranial vasculitis, myocardial infarction and sudden death have all been reported in cocaine abuse (Ricaurte and Langston, 1995). We present a fatal case with neurological and psychiatric symptoms, but without the usual cardiac and systemic signs. PMID- 11054143 TI - Symptomatic paroxysmal hemidystonia due to a demyelinating subthalamic lesion. AB - We present a case of paroxysmal hemidystonia in a patient with an isolated demyelinating lesion in the subthalamic region, involving the posterior arm of the internal capsule and extending to the subthalamic nucleus and mesencephalon, possibly due to multiple sclerosis. Compared with similar reports in the literature, in our case there was a paucity of lesions, permitting a more direct clinico-anatomical correlation. The role of the subthalamic region and basal ganglia circuitry in the genesis of symptomatic dystonia is discussed. PMID- 11054144 TI - Hemidystonia secondary to cervical demyelinating lesions. AB - Hemidystonia is usually associated with a structural lesion in the contralateral basal ganglia. We report a patient with definite multiple sclerosis, according to Poser's criteria, presenting with an acute-onset sustained left hemidystonia. Cranial T2-weighted magnetic resonance imaging (MRI) showed several hyperintense lesions in the centri semiovali and in the periventricular area without basal ganglia involvement. Moreover cervical spinal cord T2-weighted MRI showed two hyperintense lesions in the left posterolateral spine at C2 and C3, and one lesion in the right posterolateral spine at C4 levels. The hemidystonia improved completely after daily treatment with 1000 mg of methylprednisolone, and cervical MRI was performed after the improvement which showed that the lesions had become smaller and less intense. Finally we consider that the hemidystonia may be caused by the cervical spinal cord lesions of multiple sclerosis. PMID- 11054145 TI - Diagnostic challenges in combined multiple sclerosis and centronuclear myopathy. AB - The first case of combined centronuclear myopathy and multiple sclerosis is reported. The difficulties of diagnosing multiple sclerosis in patients with muscular disorders associated with the central nervous system involvement are discussed. PMID- 11054146 TI - Peripheral neuropathy associated with common variable immunodeficiency. AB - We report a patient with common variable immunodeficiency (CVID) who developed an axonal sensorimotor polyneuropathy, a hitherto unreported association to our knowledge. These conditions may be linked at the pathogenetic level, since some CVID patients are prone to the development of autoimmune disease. PMID- 11054147 TI - Lesion of the anterior branch of axillary nerve in a patient with hereditary neuropathy with liability to pressure palsies. AB - We report the case of a 30-year-old woman affected by hereditary neuropathy with liability to pressure palsies (HNPP), who developed a painless left axillary neuropathy after sleeping on her left side, on a firm orthopaedic mattress, in her eighth month of pregnancy. Electromyography (EMG) showing neurogenic signs in the left anterior and middle deltoid, and normal findings in the left teres minor, posterior deltoid and other proximal upper limb muscles, demonstrated that the lesion was at the level of the axillary anterior branch. A direct compression of this branch against the surgical neck of the humerus seems the most likely pathogenic mechanism. This is the first documented description of an axillary neuropathy in HNPP. Knowledge of its possible occurrence may be important for prevention purposes. PMID- 11054148 TI - Three-dimensional transcranial colour-coded duplex sonography of the transverse sinus. AB - The case of a patient with partial superior sagittal sinus and partial right sided transverse sinus thrombosis, and posterior fossa venous collaterals is presented. We report on the use of venous transcranial colour-coded duplex sonography (TCCS), combined with a new three-dimensional transcranial duplex data acquisition system, which allows free hand scanning of the region of interest. TCCS with three-dimensional image reconstruction allowed a more precise spatial localization of venous flow signals in the posterior fossa and facilitated the understanding of the haemodynamics of the venous collateral network. PMID- 11054149 TI - An uncommon central nervous system manifestation in systemic lupus erythematosus: the diffuse and symmetrical lesions of white matter. PMID- 11054150 TI - Meningoencephalitis and Chlamydia pneumoniae infection. PMID- 11054151 TI - Introduction: pramipexole: a novel dopamine agonist for the treatment of Parkinson's disease. PMID- 11054152 TI - Why delaying levodopa is a good treatment strategy in early Parkinson's disease. PMID- 11054153 TI - How to succeed in using dopamine agonists in Parkinson's disease. AB - Dopamine receptor agonists are assuming increased importance in the treatment of both early and advanced symptoms of Parkinson's disease (PD). However, tolerability of these drugs can be a problem. Identifying patients who are at increased risk of adverse effects is central to using dopamine agonists in PD. The newer agonists, pramipexole and ropinirole, are generally adequate without levodopa for early symptoms and carry the hope for a more acceptable profile of long-term side-effects. In the patient with advanced disease, all four dopamine agonists significantly augment the response to levodopa, which reduces the problems of motor fluctuations and drug related dyskinesia. Understanding the common pitfalls when prescribing these drugs will facilitate their safety and efficacy. PMID- 11054154 TI - Pre-clinical studies of pramipexole: clinical relevance. AB - This paper reviews the preclinical study of the novel dopamine agonist pramipexole and its use in early Parkinson's disease (PD). Emphasis will be given to those properties distinguishing this drug from other dopamine agonists, the relevance of the preclinical data to clinical trial results in early PD, and the putative neuroprotective properties of the compound. The conventional dopamine agonists are ergot-derived compounds that are most widely used as adjunctive therapies in advancing Parkinson's disease (PD). Examples of conventional agonists are bromocriptine and pergolide. Pramipexole is an aminobenzothiazole compound, recently introduced for the treatment of both early and advanced PD. Its nonergot structure may reduce the risk of side-effects, considered unique to ergot drugs, such as membranous fibrosis. Pramipexole is a full dopamine agonist with high selectivity for the D2 dopamine receptor family. This family includes the D2, D3 and D4 receptor subtypes. Pramipexole has a 5- to 7-fold greater affinity for the D3 receptor subtype with lower affinities for the D2 and D4 receptor subtypes. The drug has only minimal alpha2-adrenoceptor activity and virtually no other receptor agonism or antagonism. The optimal dopamine receptor activation for the safe and effective treatment of PD is not known. Findings in animal models and clinical studies indicate that activation of the postsynaptic D2 receptor subtype provides the most robust symptomatic improvement in PD. Given its pharmacological profile, it is not surprising that pramipexole was found to be effective in ameliorating parkinsonian signs in animal models. This therapeutic effect has been confirmed in clinical trials in both early and advanced PD. In early disease, it provides a clear reduction in the chief motor manifestations of PD and improved activities of daily living. Perhaps most striking is the large number of clinical trial patients who have remained on pramipexole monotherapy for many months. The majority of these subjects have been maintained on pramipexole for an excess of 24 months without requiring additional symptomatic treatment with levodopa. This is in contrast to the general clinical experience with older conventional agonists. Pramipexole also has a favourable pharmacokinetic profile. It is rapidly absorbed with peak levels appearing in the bloodstream within 2 h of oral dosing. It has a high absolute bioavailability of > 90% and can be administered without regard to meals. It has no significant effects on other antiparkinson drugs such as levodopa or selegiline. Its excretion is primarily renal and, thus, has little or no impact on hepatic cytochrome P450 enzymes or other related metabolic pathways. Pramipexole has also been theorized to have 'neuroprotectant' properties. Oxyradical generation is posited as a cause or accelerant of brain nigral cell death in PD. Pramipexole stimulates brain dopamine autoreceptors and reduces dopamine synthesis and turnover which may minimize oxidative stress due to dopamine metabolism. Furthermore, the compound has a low oxidation potential that may serve as an oxyradical scavenger in the PD brain. In summary, pramipexole is a new antiparkinson medication found to have unique dopamine agonist characteristics and putative neuroprotective properties. PMID- 11054155 TI - Pramipexole in the treatment of advanced Parkinson's disease. AB - Pramipexole is a novel nonergoline dopamine agonist with a preference for the dopamine D3 receptor subtype. Its efficacy and safety in the treatment of advanced Parkinson's disease has been investigated in several clinical studies. This review provides a summary of the data currently available, particularly in reference to the recent results of the European clinical phase III study and the potential tremorlytic activity of pramipexole. Interim analysis of the open-label European clinical phase III study has provided evidence of long-term efficacy and safety of pramipexole. In another study pramipexole has been shown to be significantly superior to placebo with an improvement in tremor score by 48% (vs. 13% in the placebo group). In addition to its likely usefulness in the treatment of rest tremor in Parkinson's disease, data suggest that pramipexole is of interest due to its reported low frequency of cardiovascular and gastrointestinal side-effects. However, studies comparing pramipexole with other antiparkinsonian agents would be useful to further define its benefits in the treatment of tremor dominant Parkinson's disease and to further document its favourable adverse event profile. PMID- 11054156 TI - Pramipexole in the treatment of restless legs syndrome: a follow-up study. AB - The restless legs syndrome (RLS) is a condition characterized by unpleasant limb sensations occurring at rest and associated with an irresistible urge to move. Several treatments are used to treat RLS including benzodiazepines, opioids, dopaminergic agents, clonidine and anticonvulsant drugs such as carbamazepine and gabapentine. Dopaminergic agents are now considered the treatment of choice for RLS. Levodopa is effective in treating RLS; however, several patients treated with levodopa at bedtime developed morning or late afternoon restlessness. Recently, more attention has been paid to dopamine receptor agonists. Ergoline derivatives, bromocriptine and pergolide were found effective, but require concomitant administration of domperidone, a peripheral dopamine antagonist. In a recent study, we studied the efficacy and innocuity of pramipexole, a new dopamine agonist with a higher affinity for the D3 receptor subtype of the D2 family, in a double-blind, placebo-controlled, randomized trial. Pramipexole had major effects on RLS symptoms without severe side-effects. The present study aimed to assess the long-term efficacy of pramipexole. Seven patients were treated with the drug for a mean follow-up duration of 7.8 months. Treatment was started at a dosage of 0.25 mg, and progressively increased until the optimal therapeutic effect was obtained. Home questionnaires were completed for 7 consecutive days, after one month and after a mean of 7.8 months of treatment with pramipexole, assessing leg restlessness during the daytime, in the evening, at bedtime and during the night. There was no evidence of a decrease in the therapeutic effect of pramipexole in these patients, even 7.8 months after the initiation of treatment. The optimal dosage was 0.25 mg for one patient, 0.5 mg for five patients and 0.75 mg for one patient. While there was a progressive increase in severity of leg restlessness from daytime to nighttime before treatment, a suppression of leg restlessness was observed throughout the 24 h with a single dose of pramipexole at bedtime. The remarkable efficacy of pramipexole raises the possibility that the D3 receptors of the mesolimbic system may be more specifically involved in the physiopathology of RLS. PMID- 11054157 TI - Isolation and characterization of a Lactobacillus amylovorus mutant depleted in conjugated bile salt hydrolase activity: relation between activity and bile salt resistance. AB - Growth experiments were conducted on Lactobacillus amylovorus DN-112 053 in batch culture, with or without pH regulation. Conjugated bile salt hydrolase (CBSH) activity was examined as a function of culture growth. The CBSH activity increased during growth but its course depended on bile salts type and culture conditions. A Lact. amylovorus mutant was isolated from the wild-type strain of Lact. amylovorus DN-112 053 after mutagenesis with N-methyl-N'-nitro-N nitrosoguanidine. An agar plate assay was used to detect mutants without CBSH activity. In resting cell experiments, the strain showed reduced activity. Differences between growth parameters determined for wild-type and mutant strains were not detected. Comparative native gel electrophoresis followed by CBSH activity staining demonstrated the loss of proteins harbouring this activity in the mutant. Four protein bands corresponding to CBSH were observed in the wild type strain but only one was detected in the mutant. The specific growth rate of the mutant strain was affected more by bile salts than the wild-type strain. Nevertheless, bile was more toxic for the wild-type strain. In viability studies in the presence of nutrients, it was demonstrated that glycodeoxycholic acid exerted a higher toxicity than taurodeoxycholic acid in a pH-dependent manner. No difference was apparent between the two strains. In the absence of nutrients, the wild-type strain died after 2 h whereas no effect was observed for the mutant. The de-energization experiments performed using the ionophores nigericin and valinomycin suggested that the chemical potential of protons (ZDeltapH) was involved in Lactobacillus bile salt resistance. PMID- 11054158 TI - The formation of mixed culture biofilms of oral species along a gradient of shear stress. AB - A chemostat mixed culture system was used to produce two distinct ecological states, state-1 (caries-like microcosm) and state-2 (periodontal-like microcosm). Eleven bacterial species (Streptococcus gordonii, Strep. mitis I, Strep. mutans, Strep. oralis, Actinomyces naeslundii, Lactobacillus casei, Neisseria subflava, Fusobacterium nucleatum, Porphyromonas gingivalis, Prevotella nigrescens, Veillonella dispar) were used to inoculate the planktonic system. A flow cell, designed to produce convergent flow with increasing shear stress, was attached to the chemostat system, and the resultant biofilms developed from the state-1 and state-2 microcosms along the shear stress gradient were examined and compared using image analysis and viable counts. The biofilm produced from state-1 showed a lower shear stress tolerance (0.146 Pa) than the state-2 biofilm (0.236 Pa). The biofilm compositions did not vary along the gradient of shear stress and were dependent on the initial inoculum conditions. Gram-positive species were predominant in the state-1 biofilm, while Gram-negative species were predominant in state-2. PMID- 11054159 TI - Continuous production of lacticin 3147 and nisin using cells immobilized in calcium alginate. AB - Bacteriocinogenic strains, Lactococcus lactis subsp. lactis DPC 3147 and L. lactis DPC 496, producing lacticin 3147 and nisin, respectively, were immobilized in double-layered calcium alginate beads. These beads were inoculated into MRS broth at a ratio of 1:4 and continuously fermented for 180 h. Free cells were used to compare the effect of immobilization on bacteriocin production. After equilibrium was reached, a flow rate of 580 ml h(-1) was used in the immobilized cell (IC), and 240 ml h(-1) in free-cell (FC) bioreactors. Outgrowth from beads was observed after 18 h. Bacteriocin production peaked at 5120 AU ml(-1) in both IC and FC bioreactors. However, FC production declined after 80 h to 160 AU ml( 1) at the end of the fermentation. Results of this study indicate that immobilization offers the possibility of a more stable and long-term means of producing lacticin 3147 in laboratory media than with free cells. PMID- 11054160 TI - Physiology and taxonomy of thiobacillus strain TJ330, which oxidizes carbon disulphide (CS2). AB - A bacterium (strain TJ330) capable of using carbon disulphide (CS2) as its sole energy source in an acidic environment was isolated from a peat biofilter used in experiments to remove CS2 and hydrogen sulphide (H2S) from air. Its physiology and taxonomy are described here. The strain oxidized CS2, H2S and elemental sulphur to sulphate chemolithotrophically. The rate of sulphate production was highest at pH 2. The maximum growth rate constant (micromax) using CS2 as a substrate was 3.9 x 10(-2) h(-1) (generation time 18 h) and the Monod constant (Ks) was 0.97-2.6 micromol l(-1) CS2 (74-198 microg l(-1)), corresponding to an equilibrium with 15-40 ppm CS2 in the headspace. The optimum growth temperature using elemental sulphur as a substrate was 28 degrees C. The strain bears morphological and physiological similarities to Thiobacillus thiooxidans, but the latter is incapable of oxidizing CS2. The strain TJ330 (DSM 8985) showed only 44.2 + 11.8% DNA homology with the type strain T. thiooxidans ATCC 19377, while its homology with T. ferrooxidans ATCC 23270 was 17.1 + 3.4%. The strain TJ 330 represents a high-affinity bacterium which can effectively remove low CS2 concentrations in an acid environment. These properties can be utilized in biotechnological purification applications. PMID- 11054161 TI - A membrane-immunofluorescent-viability staining technique for the detection of Salmonella spp. from fresh and processed meat samples. AB - A direct staining technique was investigated for the detection of viable Salmonella in fresh and processed meats. The technique involved overnight enrichment in BPW, extraction of Salmonella cells onto a polycarbonate membrane, followed by detection of the pathogen using anti-Salmonella monoclonal antibody coupled with an antibody linked-fluorescent stain (Texas Red) and a viability stain (Sytox Green). The technique was applied to the detection of Salm. enteritidis inoculated into broth culture or minced beef and then subjected to a variety of stresses including freezing (- 20 degrees C), heating (2 or 4 min at 56.9 degrees C), low pH (5 or 3.5) or high salt (2 or 4%). The correlation between traditional plate counts and the rapid count varied widely (r2 = 0.98 0.03), depending on the type and level of stress applied to the cells. The reason for the disparity in results obtained, and the potential application of the method as a diagnostic tool, are discussed. PMID- 11054162 TI - Direct microscopy of bacillus endospore germination in soil microcosms. AB - Antagonistic endospore-forming Bacillus spp. offer a large potential as seed inoculants for control of soil-borne pathogens. In the soil, however, inoculated Bacillus endospores may remain dormant without germination, and plant protection can therefore be inefficient and unpredictable. A method based on direct fluorescence microscopy in soil microcosms was used to determine whether low-cost organic additives incorporated into seed coating material could stimulate endospore germination. Complex organic additives supported a high level of endospore germination of the fungal antagonist Paenibacillus polymyxa CM5-5. Skim milk is a low-cost additive that may be incorporated into seed coating material for efficient induction of Bacillus endospore germination in soil. PMID- 11054163 TI - An indirect immunofluorescent antibody technique for detection and enumeration of Vibrio vulnificus serovar E (biotype 2): development and applications. AB - The applications of an indirect fluorescent antibody technique (IFAT), developed to detect and enumerate the pathogenic bacterium Vibrio vulnificus serovar E from water and clinical samples, are described. This technique proved accurate for detecting V. vulnificus, even under starvation conditions and in the non culturable state, and could differentiate this species from other bacteria which share the same habitats. The IFAT was successfully used to diagnose vibriosis from naturally- and artificially-infected eels. The overall data suggest that applying this technique properly in environmental and epidemiological/epizootiological studies could significantly increase our knowledge of this bacterium. PMID- 11054164 TI - Production and secretion of collagen-binding proteins from Aeromonas veronii. AB - Collagen-binding protein (CNBP) synthesized by Aeromonas veronii is located conserved within the subcellular fraction. The results of this study show that 98% of the total CNBP produced by Aer. veronii is present in the extracellular medium, and that the remaining CNBP is distributed either on the cell surface, within the periplasm or anchored on the outer membrane. CNBP is specifically secreted from Aer. veronii into the culture medium, because all the beta lactamase activity was located in the cells and could be released by polymixin B extraction of periplasmic proteins. CNBP was produced at growth temperatures from 12 degrees C to 42 degrees C, but not at 4 degrees C. The findings indicate that the level of CNBP in the medium increases during the exponential growth phase and reaches a maximum during the early stationary phase. There was less CNBP production in poor nutrient MMB medium than in the rich LB nutrient medium. CNBP secretion, in contrast to aerolysin secretion, was unaffected by the exeA mutation of Aer. hydrophila. It is concluded that CNBP secretion from Aer. veronii must be achieved by a mechanism different from that reported for aerolysin secretion. PMID- 11054165 TI - Anti-adhesive and antibacterial properties of a proprietary denture cleanser. AB - The antimicrobial efficacy and anti-adhesive attributes of a proprietary denture cleanser were evaluated. To determine cleanser antimicrobial efficacy, Streptococcus mutans was grown on denture acrylic strips which were then exposed to the cleanser. To evaluate anti-adhesive efficacy, the strips were treated with the cleanser and then placed in the Strep. mutans suspension. Following incubation, adhered bacteria were removed and enumerated by viable counting. Treated denture acrylic plates were also placed in a parallel-plate flow chamber and then exposed to Strep. oralis. Images of adhered bacteria were analysed to determine biofilm coverage. Biofilm removal force was quantified by increasing the flow rate between the acrylic plates. The cleanser exhibited a 100% kill against Strep. mutans adhered to the acrylic surface, and inhibited attachment of cells by 66%. The flow chamber study found that cleanser-treated denture acrylic allowed the formation of a multilayer biofilm which was easily removed by a slight increase in flow rate. PMID- 11054166 TI - Effect of packaging conditions on the growth of micro-organisms and the quality characteristics of fresh mushrooms (Agaricus bisporus) stored at inadequate temperatures. AB - Mushrooms were packed in two polymeric films (perforated and non-perforated PVC) and stored at 17 degrees C and 25 degrees C. The carbon dioxide and oxygen content inside the packages, aerobic mesophiles, Pseudomonas spp., faecal coliforms, Escherichia coli, anaerobic spores and major sensory factors (colour, texture, development stage and presence of moulds) were determined. The non perforated packages had the highest contents of CO2 (6-7%), the lowest contents of O2 (0.013-0.17%) and the most desirable quality parameters (texture, development stage and absence of moulds). Pseudomonas spp. counts were around 1 logarithmic unit lower in mushrooms packaged in non-perforated film as the O2 concentrations were lower than in perforated film. The mushrooms themselves were inoculated with an enterotoxin A-producing strain of Staphylococcus aureus, packaged in overwrapped trays and stored at 17 and 25 degrees C. Staphylococcus aureus did not grow in the samples stored at 17 degrees C. Only slight growth was observed in mushrooms packaged with non-perforated film after 1 day at 25 degrees C. No enterotoxin was detected in any package. Faecal coliform counts were <2 log cfu g(-1). Escherichia coli was not isolated in any of the samples. At 25 degrees C, counts of anaerobic spores of around 2 log cfu g(-1) were detected in those mushrooms packaged in non-perforated film. PMID- 11054167 TI - Molecular analysis of tetracycline resistance in Salmonella enterica subsp. enterica serovars Typhimurium, enteritidis, Dublin, Choleraesuis, Hadar and Saintpaul: construction and application of specific gene probes. AB - A total of 65 epidemiologically unrelated tetracycline-resistant isolates of the six Salmonella enterica subsp. enterica (Salm.) serovars Dublin, Choleraesuis, Typhimurium, Enteritidis, Hadar and Saintpaul were investigated for the presence of tetracycline resistance genes. For this, specific gene probes of the tetracycline resistance genes (tet) of the hybridization classes A, B, C, D, E and G were constructed by cloning PCR-amplified internal segments of the respective tet structural genes. These gene probes were sequenced and used in hybridization experiments with plasmid DNA or endonuclease digested whole cell DNA as targets. Only tet(A) genes were detected on plasmids in all Salm. Dublin isolates as well as in single isolates of Salm. Choleraesuis and Salm. Typhimurium. Genes of the hybridization classes B, C, D and G, but also in some cases those of class A, were located in the chromosomal DNA of the corresponding Salmonella isolates. Restriction fragment length polymorphisms (RFLPs) of tet gene carrying fragments were detected in chromosomally tetracycline-resistant isolates. These RFLPs might represent valuable additional tools for the identification and characterization of tetracycline-resistant Salmonella isolates. PMID- 11054168 TI - Enrichment and isolation of non-specific aromatic degraders from unique uncontaminated (plant and faecal material) sources and contaminated soils. AB - Microbial analysis of contaminated soil and uncontaminated plant and faecal material resulted in the enrichment of a number of microbial communities capable of utilizing a range of environmental pollutants. Growth was observed on polycyclic aromatic hydrocarbons, polychlorinated biphenyls, heterocyclic aromatic compounds and organochlorine pesticides. However, none of the communities could grow on pentachlorophenol. Pure cultures were isolated from microbial communities using phenanthrene and pyrene as the sole carbon and energy source. Isolates were also obtained using DDT, DOH, DBH and PCPA when peptone was supplemented to the medium. Strain AJR39,504, isolated using DDT and peptone, could not be positively identified on the basis of substrate utilization tests. However, it most closely resembled Stenotrophomonas maltophilia (0.424 similarity) using the Microlog 3 database software. Isolate AJR39, 504 could also grow on polycyclic aromatic hydrocarbons, chlorinated- and nitro-aromatic compounds. In addition, the degradation of DDT (100 mg l(-1)) by isolate AJR39,504 resulted in a 35% decrease in DDT concentration after 28 days with a concomitant increase in DDD concentration. PMID- 11054169 TI - An extracellular sensor and an extracellular induction component are required for alkali induction of alkyl hydroperoxide tolerance in Escherichia coli. AB - Escherichia coli K12 transferred from pH 7.0 to pH 9.0 gains alkylhydroperoxide (AHP) tolerance. The aim here was to establish whether extracellular components (ECs) are needed for such induction. Therefore, the effects of removing ECs during incubation at pH 9.0 were tested and the abilities of culture filtrates to induce tolerance were examined. First, AHP tolerance did not appear, at pH 9.0, if cultures were subjected to continuous filtration or dialysis, against the same medium, suggesting that an EC might be needed. Second, neutralized filtrates from pH 9.0-grown cultures induced tolerance at pH 7.0, and these filtrates were inactivated by dialysis, filtration or heating but not by protease. Thus, pH 9.0 filtrates have a small non-protein extracellular induction component (EIC), which acts as an alarmone, 'warning' cells of stress and preparing them to resist it. Filtrates from pH 7.0-grown cultures did not induce AHP tolerance at pH 7.0 but if incubated at pH 9.0 without organisms, gained such ability. It is proposed that pH 7.0 filtrates have an EIC precursor (termed an extracellular sensing component, ESC), which senses alkaline pH, and is converted by it to the EIC. The ESC in pH 6.0 filtrates was distinct from that in pH 7.0 filtrates; there may be several oligomeric (or conformational) forms of this ESC. As the EIC is small, it can diffuse away from the alkalinized region and induce tolerance in unstressed organisms. PMID- 11054170 TI - A genetic study of natural flor strains of Saccharomyces cerevisiae isolated during biological ageing from Sardinian wines. AB - In this study, three flor strains of Saccharomyces cerevisiae were genetically characterized. They were isolated from biofilms on Sardinian sherry-like wines produced at family-run wineries where pure cultures of yeasts were not used. The study aimed to investigate the life cycle of these naturally-occurring flor strains, using a genetic procedure supplemented by analysis of subsequent meiotic generations. A semi-homothallic life cycle was found in three strains that could be helpful in a genetic improvement programme. PMID- 11054171 TI - Virulence plasmid (pYV)-associated susceptibility of Yersinia enterocolitica to chlorine and heavy metals. AB - This study was aimed at elucidating the role of virulence plasmid (pYV) in the susceptibility of Yersinia enterocolitica to bactericidal agents such as chlorine and heavy metals. Plasmid-bearing (pYV+) Y. enterocolitica was less susceptible to the antimicrobial action of chlorine and heavy metals compared with the isogenic plasmidless (pYV-) derivative. This difference was, however, observed only with bacteria cultured at 25 degrees C. pYV-associated susceptibility apart, cells cultured at 37 degrees C were also found to be less susceptible to the antimicrobial action of these agents. The results indicate that the susceptibility of Y. enterocolitica to these agents was influenced both by the presence of the virulence plasmid and the temperature at which the cells were cultured. PMID- 11054172 TI - Evaluation of 16s rRNA and cellular fatty acid profiles as markers of human intestinal bacterial growth in the chemostat. AB - Chemostats were used to study the effects of carbon and nitrogen limitation and specific growth rate on 16S rRNA synthesis and cellular fatty acid (CFA) profiles in four human intestinal bacteria (Bacteroides thetaiotaomicron, Bifidobacterium adolescentis, Clostridium bifermentans and Cl. difficile). Cellular fatty acid synthesis varied with dilution rate and nutrient availability in different species, but these cellular constituents were relatively stable phenotypic characteristics in Bact. thetaiotaomicron, where branched chain and hydroxy CFA were good taxonomic markers. Conversely, CFA in the Gram-positive bacteria varied markedly with changes in growth environment. For example, in chemostats, cyclopropane CFA were only synthesized in Cl. bifermentans and Cl. difficile under N-limited conditions. Similarly, Dimethyl acetal (DMA) fatty acids in Bif. adolescentis were primarily produced during N-limited growth, and this was inversely related to dilution rate. At low growth rates, 16S rRNA concentrations (microg rRNA per ml culture) correlated with viable bacterial counts, but were more closely related to specific growth rate when expressed as a function of cell mass (microg rRNA per mg dry weight bacteria). However, this did not reveal differences in bacterial population size and rRNA concentration in C-limited cultures. Thus, at low dilution rates, C limitation strongly reduced rRNA synthesis in Cl. bifermentans, despite viable cell counts being similar to those in N-limited cultures. These results indicate that, while 16S rRNA is a useful indicator of microbial activity, cell growth rate does not necessarily relate to rRNA concentration under all nutritional conditions. Consequently, bowel habit and diet will affect both CFA and rRNA content in bacteria isolated from intestinal samples, and this should be taken into consideration when interpreting such data measurements. PMID- 11054173 TI - Extrusion of wheat or sorghum and/or addition of exogenous enzymes to pig diets influences the large intestinal microbiota but does not prevent development of swine dysentery following experimental challenge. AB - A study was made of dietary influences on the large intestinal microbiota of pigs and on the incidence of swine dysentery (SD) after experimental infection with Brachyspira hyodysenteriae, the aetiological agent of SD. Animals were fed diets based either on wheat (expts 1 and 2) or sorghum (expt 2). Grains were ground and fed either raw or after high temperature and pressure extrusion and/or after addition of exogenous enzymes to the whole diet to reduce the starch and soluble non-starch polysaccharides available for fermentation in the large intestine. Limiting fermentation creates conditions that apparently reduce the incidence of SD after infection with B. hyodysenteriae. The diets were fed to weaned pigs for 4-6 weeks, then half the animals on each diet were killed and gut samples collected for microbiology. The treatments had little effect on bacterial numbers. In expt 1, dietary extrusion of wheat reduced lactobacilli in the large intestine. Addition of enzymes to extruded wheat-based diets in expt 2 reduced facultative anaerobes and increased non-sporing anaerobes. Addition of enzymes to a raw sorghum diet in expt 3 decreased numbers of facultative anaerobes, while extrusion of sorghum increased total anaerobes. Bacteroides spp. and Fusobacterium spp., which act in synergy with B. hyodysenteriae in SD, were isolated at a higher percentage in pigs fed the untreated wheat diet than in pigs fed the treated wheat diets. Following experimental infection the incidence of SD amongst pigs fed treated wheat diets was slightly lower than those fed the untreated diet, but with sorghum-based diets the opposite was found. Overall, the different dietary treatments used did not significantly reduce SD. PMID- 11054174 TI - Characterization of Sphingomonas isolates from Finnish and Swedish drinking water distribution systems. AB - Sphingomonas species were commonly isolated from biofilms in drinking water distribution systems in Finland (three water meters) and Sweden (five water taps in different buildings). The Sphingomonas isolates (n = 38) were characterized by chemotaxonomic, physiological and phylogenetic methods. Fifteen isolates were designated to species Sphingomonas aromaticivorans, seven isolates to S. subterranea, two isolates to S. xenophaga and one isolate to S. stygia. Thirteen isolates represented one or more new species of Sphingomonas. Thirty-three isolates out of 38 grew at 5 degrees C on trypticase soy broth agar (TSBA) and may therefore proliferate in the Nordic drinking water pipeline where the temperature typically ranges from 2 to 12 degrees C. Thirty-three isolates out of 38 grew at 37 degrees C on TSBA and 15 isolates also grew on blood agar at 37 degrees C. Considering the potentially pathogenic features of sphingomonas, their presence in drinking water distribution systems may not be desirable. PMID- 11054175 TI - Drug uptake via nutrient transporters in Trypanosoma brucei. PMID- 11054176 TI - A new bacteriophage, VHML, isolated from a toxin-producing strain of Vibrio harveyi in tropical Australia. AB - Some strains of Vibrio harveyi are known to be pathogenic for fish and many invertebrates including crustaceans. Despite their importance, their modes of virulence have yet to be fully elucidated. Here, we present a previously unreported bacteriophage extracted from a toxin-producing strain of V. harveyi isolated from moribund prawn larvae in tropical Australia. Classification into the family Myoviridae was based upon morphological characteristics (an icosahedral head, a neck/collar region and a sheathed rigid tail) and nucleic acid characteristics (double-stranded linear DNA). We have termed the bacteriophage VHML (Vibrio Harveyi Myovirus Like). VHML is a temperate bacteriophage that has a narrow host range and shows an apparent preference for V. harveyi above other vibrios (63 Vibrio isolates tested) and other genera (10 other genera were tested). The conventional methods for phage concentration and extraction of nucleic acids from phage particles were not efficient and the alternative methods that were used are discussed. PMID- 11054177 TI - Efficacy of a commercial polymerase chain reaction-based assay for detection of Salmonella spp. in animal feeds. AB - Salmonellosis is a cyclic problem in the food industry, to which animal feed has been contributory. Current conventional methods of Salmonella spp. detection require 96 h for detection and confirmation. With modern and just-in-time production schedules, a 96-h hold represents a significant expense in storage and decontamination. The commercially available assay, 'BAX for Screening/Salmonella' (BAX), is based on the principle of the polymerase chain reaction and may represent a significant decrease in assay time. Seven fresh feed formulations, two fresh feed ingredients, seven stored feeds and two stored feed ingredients were artificially contaminated with a primary poultry isolate of Salmonella typhimurium and analysed by conventional and BAX methodology. The results of BAX agreed with conventional plating results for 16 of 18 samples spiked with 1200 cfu 10 g(-1) of feed and 13 of 18 samples spiked with 40 cfu 10 g(-1) of feed. Indigenous Salmonella spp. were detected in five of eight samples of poultry diets by conventional methods. With BAX, Salmonella spp. could not be detected in any of the samples after only 7 h of enrichment but could be detected in two dietary samples after 13 h of enrichment and four dietary samples after 24 h of enrichment. Specific sequences of salmonella DNA that were extracted from poultry diets could be detected with BAX. PMID- 11054178 TI - Detection of a novel campylobacter cytotoxin. AB - The culture filtrates from 10 Campylobacter species were screened for the presence of cytotoxins on a variety of selected tissue culture cell lines. Some Campylobacter jejuni strains showed no effects on tissue culture cell lines compared with other C. jejuni strains, especially C. jejuni 81116, which consistently produced a cytotoxin that was lethal to tissue culture cells. It was observed that CHO cells were the most sensitive cell line in detecting campylobacter cytotoxins. Samples containing the culture filtrate of C. jejuni 81116 prepared at various growth stages were used to determine the subcellular location of the cytotoxin. This C. jejuni 81116 cytotoxin appears to be a heat stable toxin that is secreted from the cell during stationary phase; cytotoxin activity can be abolished with proteolytic enzymes. PMID- 11054179 TI - vCJD - predicting the future? AB - The recent emergence of variant Creutzfeldt-Jakob disease (vCJD) in the UK, and demonstration that vCJD is caused by the same prion strain that causes bovine spongiform encephalopathy, have led to concerns about the possibility of a human epidemic. Although only 79 cases of vCJD have occurred to date, it is likely that hundreds of thousands of infected cattle entered the human food chain in the late 1980s and early 1990s, and the average incubation period of vCJD is unknown. Mathematical models have not yet been able to give useful predictions of future numbers of cases, and in the absence of a blood test for vCJD, current attempts to reduce uncertainties about future numbers of cases are based on the accumulation of PrPSc in lymphoreticular tissues. Extensive lymphoreticular PrPSc accumulation has been seen in all cases of symptomatic vCJD so far examined, and in one case 8 months prior to the onset of symptoms. Animal models of prion disease suggest that lymphoreticular involvement occurs early in the incubation period and reliably predicts future neurological disease. Based on these data, large scale anonymous studies looking for PrP accumulation in surgically removed tonsillectomy and appendicectomy specimens are underway. Examination of the first 3000 specimens has not revealed any positive samples, but at the moment the significance of negative findings is uncertain. It is anticipated that by the time these studies are complete more data will be available on how early PrP can be demonstrated in lymphoreticular tissue in vCJD, which together with the results from examination of further samples, will allow some comment as to the likelihood of a large human vCJD epidemic. PMID- 11054180 TI - The ethics of reusing archived tissue for research. AB - Pathologists have been establishing archives of human organs and tissue for research use for many years now. Controversy has arisen recently over these collections, particularly with regard to the right of patients or relatives to consent to removal and retention of tissue, genetic research using stored tissue samples, and commercial exploitation of tissue collections and genetic material. This paper discusses the ethics of reusing existing archives of tissue. New archives are established under much more stringent conditions than in the past. What rules should apply to existing archives? Guidelines to regulate such use are useful, but face serious difficulties in balancing the variety of public and private interests relating to tissue banking. Consent cannot be obtained retrospectively, but public trust can be established by open acknowledgement of the evolution of ethical standards and strict adherence to current best practice. Guidelines and standards vary from country to country, but ethical principles should not. The implications of this view for pathologists worldwide are discussed. PMID- 11054181 TI - Oedema and glial cell involvement in the aged mouse brain after permanent focal ischaemia. AB - This study examines the effect of age on oedema and brain swelling, and associated glial cell involvement on the size of the lesion in two models of permanent, focal cerebral ischaemia. Ischaemia was induced in male C57BL/Icrfat mice (4-6 and 26-31-month-old) by middle cerebral artery (MCA) occlusion using either electrocoagulation after craniotomy (MCA/craniotomy), or by an intraluminal filament through the carotid artery (MCA/icf). Twenty-four hours after inducing ischaemia, brain swelling and lesion size were measured in young and aged mice, and cerebral oedema by wet/dry brain weights. Histopathology and immunocytochemistry were performed on a separate set of perfusion fixed brains. The MCA/icf technique produced a significantly larger lesion than MCA/craniotomy in both age groups. The percentage of water taken into the brain was significantly greater after MCA/icf, with aged mice showing the greatest increase. When lesion size was corrected for brain swelling there was no age related increase in the size of the lesion. The numbers of microglia and astroglia increased significantly in the parietal cortex of aged control animals, and there were qualitative differences in the glial response between the two stroke models. This study emphasizes the importance of age in models of permanent focal ischaemia, with oedema clearly being a significant factor. Differ-ences in the responsiveness of the glial cell population with age may be of fundamental importance in the progress of ischaemic brain damage. PMID- 11054182 TI - Fas and Fas-L expression in Huntington's disease and Parkinson's disease. AB - The Fas/Fas-L signalling system plays a role in the control of cell death and the survival of lymphocytes, in the regulation of the immune system, and in the progression of autoimmune diseases. Studies in the nervous system have shown Fas/Fas-L activation in multiple sclerosis and in various paradigms leading to neuronal death. Enhanced Fas and Fas-L expression has also been documented in astrocytomas and glioma cell lines. However, little is known about the possible implication of Fas/Fas-L signals in primary human neurodegenerative diseases. In an attempt to gain understanding of the mechanisms commanding cell death and neurone loss in Huntington's disease (HD) and Parkinson's disease (PD), Fas and Fas-L expression has been examined in the brains of patients with HD and PD with Western blotting and immunohistochemistry. Fas and Fas-L expression levels are reduced in the caudate and putamen, but not in the parietal cortex, in HD, as revealed in Western blots. Moreover, Fas and Fas-L immunoreactivity is reduced in striatal neurones in HD. Fas and Fas-L immunoreactivity is also decreased in neurones of the substantia nigra pars compacta in PD. Reduced Fas and Fas-L expression is observed equally in Lewy body-bearing and non-Lewy body-bearing neurones. Yet increased Fas and Fas-L immunoreactivity occurs in normal astrocytes in control brains and in reactive astrocytes in diseased brains. The meaning of increased Fas and Fas-L expression in astrocytes is still unclear. However, the present results suggest that Fas/Fas-L signals are minimized in sensitive neurones in HD and PD. PMID- 11054183 TI - Connective tissue growth factor is expressed by a subset of reactive astrocytes in human cerebral infarction. AB - Connective tissue growth factor (CTGF), a transforming growth factor (TGF)-beta1 downstream mediator, is a secreted cell matrix-inducing peptide involved in both tissue regeneration mechanisms, such as wound repair, and also in aberrant deposition of extracellular matrix. The present study reports CTGF expression by cells associated with matrix deposition and glial scar formation in human cerebral infarction. CTGF was localized by immunohistochemistry in 17 brains of patients after focal infarction and in three neuropathologically normal control brains. CTGF expression was selectively localized to the cytoplasm of stellate reactive astrocytes. Compared to peripheral areas and brain controls without neuropathological findings, the total number CTGF+ astrocytes was significantly higher (P < 0.0001) in border zones adjacent to the core, corresponding to the penumbra. These numbers were significantly increased at day 1 and day 3 and remained persistently elevated up to several months post-infarction (P < 0.0001). The restricted expression and accumulation of CTGF+ reactive astrocytes adds convincing evidence for CTGF participation in the gliotic astrocyte CNS injury response involved in glial scar formation. CTGF can be considered a sensitive marker of early human astrocyte activation and a possible target for pharmacological intervention of aberrant matrix deposition. PMID- 11054184 TI - Molecular genetic analysis of archival gliomas using diagnostic smears. AB - Investigation of the clinical significance of genetic alterations in gliomas requires molecular genetic analysis using samples from retrospective or prospective clinical studies. However, diagnostic tissue is often severely limited and because of fixation, paraffin-embedded tissues (PET) contain degraded DNA. Intra-operative cytological preparations (smears) archived after diagnosis may represent an additional source of clinical material for genetic analysis. In this study, tissue samples were obtained by precision microdissection of archived diagnostic smears from 20 cases (1961-1999). All samples produced polymerase chain reaction (PCR) products for the beta globin gene, but the most recent samples amplified best and gave longer amplimers. For six cases, direct comparison was made between samples microdissected from smears and the corresponding PET. Samples from smears showed improved PCR performance and similar alleles on microsatellite marker analysis. One case, with smears of uninvolved cortex and tumour tissue available for microdissection, showed allelic imbalance at 10q23 on the basis of the smear results alone. PCR products from smears were shown to be suitable for direct sequence analysis (p53 gene). A PTEN mutation, found previously in an anaplastic astrocytoma by analysis of PET, was detected in the corresponding diagnostic smear. The results of this study indicate that tissue samples microdissected from diagnostic intra-operative cytological preparations may be suitable for molecular genetic analysis of gliomas. PMID- 11054185 TI - Hypoglycaemia is a cause of axonal injury. AB - Axonal injury as demonstrated immunohistochemically is increasingly being recognized at post-mortem in patients who have been unconscious, and in some cases the cause of the coma may not be immediately apparent. Considerations include microscopical diffuse traumatic axonal injury and axonal injury associated with a range of metabolic encephalopathies. In this study, extensive neurohistological examination was undertaken in 13 patients in whom coma was attributed to hypoglycaemia and in whom neurohistological examination had revealed varying degrees of widely distributed neuronal necrosis: in five of these cases there was also evidence that the intracranial pressure had been high with internal hernation. It is concluded that a significant amount of axonal injury found in these 13 cases can be attributed to hypoglycaemia per se although the amount and distribution of the axonal damage is altered in the presence of raised intracranial pressure. However, in some cases axonal damage is seen in the absence of an elevated intracranial pressure and in one case its distribution closely mimicked that seen in microscopical diffuse traumatic axonal injury. This further demonstrates that not all axonal pathology is traumatic, and that adequate sampling and care in interpretation of Abeta-PP staining is required in forensic practice. PMID- 11054186 TI - Blood-brain barrier disruption in simian immunodeficiency virus encephalitis. AB - Infected monocyte-derived macrophages (MDM) are thought by some investigators to play a central role in the neuropathogenesis of human immunodeficiency virus encephalitis (HIVE). It was recently proposed that these cells gain access to the central nervous system (CNS) through disruptions in blood-brain barrier (BBB) tight junctions, which occur in HIVE in association with accumulation of activated, HIV-1-infected, perivascular macrophages and serum protein extravasation (Am J Pathol 1999, 155: 1915-27). The present study tested this hypothesis in basal ganglia tissue from simian immunodeficiency virus (SIV) infected macaques with encephalitis by examining vessels for immunohistochemical alterations in the tight junction-associated proteins, occludin and zonula occludens-1 (ZO-1). Compared to non-infected macaques and SIV-infected macaques without encephalitis, cerebral vessels from macaques with SIVE showed fragmentation and decreased immunoreactivity for both tight junction proteins. These alterations were associated with accumulation of perivascular macrophages and aberrant occludin and ZO-1 immunoreactivity within these cells. In addition, perivascular extravasation of fibrinogen, a plasma protein, and a change from a strong linear staining pattern to a more irregular pattern of glucose transporter isoform-1 (GLUT-1), a metabolic BBB marker, were observed in regions with vascular tight junction protein alterations. These findings demonstrate that tight junction disruption occurs in SIVE in association with perivascular macrophage accumulation. While it cannot be ascertained from these studies whether such changes precede macrophage infiltration, or are secondary to the chronic presence of macrophages around cerebral vessels, disruptions in BBB integrity could serve as portals for additional accumulation of perivascular macrophages in SIVE. PMID- 11054187 TI - Clinicopathological phenotype of codon 129 valine homozygote sporadic Creutzfeldt Jakob disease. AB - The naturally occurring polymorphism at codon 129 of the human prion protein gene (PRNP) influences susceptibility to sporadic Creutzfeldt-Jakob Disease (CJD); the majority of the patients are methionine homozygotes at this locus, while valine homozygotes represent only 10% of cases. The aim was to study the clinical and neuropathological phenotype of sporadic CJD in valine homozygotes, to estimate the reliability of current clinical diagnostic criteria, and to identify any consistent and distinct features. Twelve cases of sporadic CJD with a codon 129 valine homozygote genotype were identified at the National CJD Surveillance Unit in Edinburgh. In addition to a retrospective clinical analysis, tissue blocks were stained by conventional techniques and by immunocytochemistry for prion protein. Frozen brain tissue was available from five cases for Western blot analysis of PrPRES, which in all cases showed a type 2 mobility. The cases included four males and eight females, average age 63.6 years, with a mean duration of illness of 6 months. Eleven patients presented with ataxia, and none had the characteristic EEG changes found in sporadic CJD. The neuropathological phenotype comprised spongiform change and prion protein immunopositivity most marked in the subcortical grey matter and cerebellum, prion protein positive plaque-like deposits in all regions, laminar deposition of prion protein in the cerebral cortex, and hippocampal involvement (which is seldom reported in sporadic CJD). In conclusion, these cases exhibited a fairly uniform phenotype, which is relatively distinct from sporadic CJD in methionine homozygotes, and thus diagnosis may be difficult using existing clinical criteria. PMID- 11054188 TI - Temporal lobe destruction with microvascular dissections following irradiation for rhinopharyngeal carcinoma. PMID- 11054189 TI - Primary progressive aphasia with focal glial tauopathy. PMID- 11054190 TI - Correlation of magnetic resonance imaging findings and histopathology of lesion distribution of spinal cord sarcoidosis at post-mortem. PMID- 11054191 TI - Subependymoma adjacent to a complex tail bud malformation: more than just a coincidence? PMID- 11054192 TI - Traumatic or diffuse axonal injury? Author's response PMID- 11054193 TI - Randomized trial of inhaled flunisolide versus placebo among asthmatic patients discharged from the emergency department. AB - STUDY OBJECTIVE: Inhaled corticosteroids (ICs) improve airflow and decrease symptoms in patients with chronic asthma. We examined whether high-dose inhaled flunisolide would have similar benefits after an emergency department visit for acute asthma. METHODS: Over a 16-month period at one inner-city ED, we documented 551 eligible patients (acute asthma; age 18 to 50 years; no ICs in past week; no oral corticosteroids in past month; and peak expiratory flow rate [PEFR] <70% of predicted value after first beta-agonist treatment); 104 patients agreed to participate. At ED discharge, all patients were given prednisone 40 mg/d for 5 days and inhaled beta-agonists as needed and were randomly assigned to receive high-dose inhaled flunisolide 2 mg/d (n=51) or placebo (n=53). Patients were telephoned daily and asked to return for PEFR measurement at 3, 7, 12, 21, and 24 days. RESULTS: Despite precautions, 28% (16 receiving flunisolide and 13 receiving placebo) of patients were completely lost to follow-up, 2 patients had only one follow-up (day 3), 2 patients receiving flunisolide withdrew because of medication-related bronchospasm, and 4 patients in each group experienced relapse. Among the 63 remaining patients, we found no difference between flunisolide and placebo at day 24 follow-up in percent predicted PEFR (87% versus 83% on day 24, P =.36; difference 4%, 95% confidence interval [CI] -5% to 13%). Nocturnal wheezing and nocturnal albuterol inhaler use was higher among patients receiving flunisolide than those receiving placebo on day 24 (48% versus 18% for nocturnal wheezing, P =.01; mean difference 30%, 95% CI 11% to 49%; 3.8 versus 1.4 nocturnal albuterol puffs, P =.03; mean difference 2.4 puffs, (95% CI 0.2 to 4). Levels of dyspnea, cough, and overall well-being were similar between the flunisolide and placebo groups. CONCLUSION: Addition of high-dose inhaled flunisolide to standard therapy does not benefit inner-city patients with acute asthma in the first 24 days after ED discharge. Airway inflammation during acute asthma may require higher doses or more potent anti-inflammatory agents. PMID- 11054194 TI - The bronchodilator effect of intravenous glucagon in asthma exacerbation: a randomized, controlled trial. AB - STUDY OBJECTIVE: Glucagon is a rapid-acting smooth muscle relaxant with a short half-life. Previous studies suggested glucagon may have bronchodilator effects. We sought to determine whether intravenous glucagon produces clinically important immediate bronchodilation in emergency department patients with asthma exacerbation. METHODS: We conducted a randomized, double-blind, placebo controlled study at 2 university-affiliated community teaching hospital EDs (annual census 90,000). ED patients 18 to 50 years old with asthma exacerbation and peak expiratory flow rate (PEFR) less than 350 L/min were eligible. Exclusion criteria were need for intubation, chronic obstructive pulmonary disease, diabetes mellitus, insulinoma, pheochromocytoma, pregnancy, lactation, or current oral steroid treatment. Patients were randomly assigned to receive glucagon 0.03 mg/kg or an equivalent volume of saline solution intravenously. At 10 minutes, PEFR was measured and all patients began standardized albuterol therapy. Successful bronchodilation was a PEFR increase of 60 L/min at 10 minutes. RESULTS: Success occurred in 2 (9.5%) of 21 glucagon-treated patients and 3 (12%) of 25 placebo-treated patients (95% confidence interval [CI] for difference of 2.5% [-20.4% to 15. 4%]). Mean PEFR improvement for glucagon was 2 L/min versus 9 L/min for placebo (95% CI for difference of -7 L/min [-36 L/min to 23 L/min]). CONCLUSION: Glucagon alone provided no clinically important immediate bronchodilation in ED patients with asthma exacerbation. PMID- 11054195 TI - The utility of anoscopy and colposcopy in the evaluation of male sexual assault victims. AB - STUDY OBJECTIVE: We sought to compare the use of anoscopy and colposcopy in examinations of male sexual assault victims and to characterize the demographics of male sexual assault victims. METHODS: This is a case series of 67 male sexual assault victims evaluated over an 8-year period by the Sexual Assault Forensic Examination team. The setting is a university-based emergency department serving as the primary site for examination of sexual assault victims by trained nurse practitioners and physician's assistants. Police and victims' advocates are available at the time of the examination. Anoscopy was done routinely over the entire study period in all patients with any anal penetration or involvement. Colposcopy use started in 1994 to magnify and take pictures. Patients were categorized into 2 groups. Group 1 consisted of subjects receiving only anoscopy, and group 2 consisted of subjects receiving initial colposcopy. Anoscopy in group 1 and colposcopy in group 2 were compared for positive results. A positive result was defined as an additional finding to those obtained by means of gross examination by using the test being evaluated (anoscopy versus colposcopy). Colposcopy and anoscopy were also compared among the subjects receiving both tests. Groups were compared by using a Pearson chi(2) test. RESULTS: Sixty-seven male sexual assault victims were evaluated between 1991 and 1998. The average age was 26+/-8 years, and the distribution of races was 30% black, 62% white, and 8% Hispanic. Results of gross examination were positive in 42 (63%) subjects. Four patients did not receive either anoscopy or colposcopy. Of the remaining 63, 25 patients had anoscopy only (group 1), and 38 patients had initial colposcopy (group 2). There were no significant differences in age, race, or rate of positive gross examination results between groups. Findings in addition to those obtained by means of gross examination were revealed by means of anoscopy in 8 (32%) of 25 subjects in group 1 and colposcopy in 3 (8%) of 38 subjects in group 2 (P =.03, difference 24%, 95% confidence interval 4% to 44%). In the 36 subjects who had both examinations, the gross examination revealed at least one finding in 22 (61%). The combination of anoscopy and colposcopy yielded positive findings in 17 subjects, including 4 subjects who had no findings on gross examination (increasing the positive rate to 26/36 [72%]). CONCLUSION: In male sexual assault victims with anal penetration, anoscopy is significantly better for gathering evidence than is colposcopy. The addition of colposcopy and anoscopy increased the rate of cases with positive findings from 61% to 72%. These 2 methods together may be a valuable adjunct in gathering evidence of damage. PMID- 11054196 TI - Prediction of hospital utilization among elderly patients during the 6 months after an emergency department visit. AB - STUDY OBJECTIVE: A simple screening tool, Identification of Seniors at Risk (ISAR), developed for administration in the emergency department for patients 65 years and older, predicts adverse health outcomes during the 6 months after the ED visit. In this study, we investigated whether the ISAR tool can also predict acute care hospital utilization in the same population. METHODS: Patients 65 years and older who visited the EDs of 4 acute care Montreal hospitals during the weekday shift over a 3-month period were enrolled. At the initial (index) ED visit, 27 self-report screening questions (including the 6 ISAR items) were administered. The number of acute care hospital days during the 6 months after the index visit were abstracted from the provincial hospital discharge database. High utilization was defined as the top decile of the distribution of acute care hospital days. RESULTS: Among 1,620 patients with linked data, a score of 2+ on the ISAR tool predicted high hospital utilization with a sensitivity of 73% and a specificity of 51%; the area under the receiver operating characteristic curve was 0.68. The ISAR tool also performed well in subgroups defined by disposition (admitted versus discharged) and by age (65 to 74 years versus 75 years and older). CONCLUSION: The ISAR tool, a 6-item self-report questionnaire, can be used in the ED to identify elderly patients who will experience high acute care hospital utilization as well as adverse health outcomes. PMID- 11054197 TI - A survey of academic departments of emergency medicine regarding operation and clinical practice. AB - STUDY OBJECTIVE: To survey academic departments of emergency medicine concerning their operation and clinical practice. METHODS: A survey was mailed to the chairs of all 56 academic departments of emergency medicine in the United States requesting information concerning operations and clinical activity in budget year 1997-1998 compared with 1995-1996. These results were then compared with a similar survey conducted in the fall of 1996, examining the 1995-1996 academic year compared with the 1994-1995 academic year. RESULTS: Forty-one (73%) academic departments of emergency medicine responded. For 1997-1998, compared with 1995 1996, 24 (59%) academic departments of emergency medicine reported an increase in emergency department patient volume; 10 (24%) reported a decrease. Twenty-four (51%) academic departments of emergency medicine reported an increase in ED patient severity, whereas 7 (15%) reported a decrease. Twenty-five (61%) academic departments of emergency medicine reported an increase in net clinical revenue for emergency medicine services, and 9 (22%) reported a decrease. Only 9 (22%) academic departments of emergency medicine reported other academic departments within their university/medical center aggressively directing patients away from the ED compared with 14 (30%) in the previous study. The percentage of academic departments of emergency medicine using midlevel providers remained essentially the same over time (68% versus 66%). In both studies, midlevel providers were used most commonly in a fast-track setting. Only 37% of academic departments of emergency medicine reported having an observation unit; staffing in all cases was by emergency physicians. Since the last survey, 38 (93%) academic departments of emergency medicine reported their medical center or hospital negotiating with managed care organizations to provide services. Unfortunately, only 41% of chairs were involved in these discussions. Between January 1, 1997, and the 1998 fall survey, 29% of academic departments of emergency medicine reported their university merging with another university system, and 19% reported such mergers being discussed. Similarly, between January 1, 1997, and fall 1998, 22% of academic departments of emergency medicine reported their institution merging with a private entity, whereas 16% reported ongoing discussions. CONCLUSION: Academic departments of emergency medicine have experienced some encouraging trends: an increase in ED patient volume, patient severity, and net clinical revenue during the study period. Midlevel providers continue to be used primarily in fast-track areas of EDs. An area of potential growth for academic departments of emergency medicine is observation medicine, because only one third of academic departments of emergency medicine have such a unit. Academic medical centers have experienced a significant increase in merger activity during the study period. PMID- 11054198 TI - Does ethanol affect the reliability of pelvic bone examination in blunt trauma? AB - STUDY OBJECTIVE: We sought to determine whether the reliability of clinical evaluation for pelvic bone fracture after trauma is compromised by a serum ethanol level of 100 mg/dL or greater. METHODS: This is a retrospective case control study of trauma registry patients presenting from October 1, 1995, to March 31, 1997, to an urban level I trauma center. Inclusion criteria were as follows: blunt trauma, age 13 years or older, and Glasgow Coma Scale score of 13 or greater. Exclusion criteria were as follows: isolated penetrating injury and suspected spinal injury. Patients were separated into 2 groups: those with an ethanol level of 100 mg/dL or greater, and those with an ethanol level of less than 100 mg/dL. Physician performance in clinical identification of pelvic bone fracture by using a complaint of pain, pelvic tenderness, with or without deformity, was compared between the 2 groups. RESULTS: Seven hundred sixty-three patients met inclusion criteria. Fifty-five (7. 2%) patients had a pelvic fracture, 75% of which were isolated acetabulum or pubic ramus fractures. Two hundred six control patients without pelvic fractures were randomly selected. The sensitivity and specificity of the complaint of pain and tenderness, deformity, or both for identification of a pelvic fracture was not significantly different between the ethanol groups. Five (9%) of 55 patients with pelvic fractures had neither a complaint of pain nor bony tenderness or deformity on examination. This was not statistically associated with an ethanol level of 100 mg/dL or greater (P =1.000). CONCLUSION: In our study, clinical evaluation for pelvic fracture in trauma patients with a Glasgow Coma Scale score of 13 or greater was not compromised by an ethanol level of 100 mg/dL or greater. The most common reason for clinically missed pelvic fractures was the presence of additional painful distracting injuries. PMID- 11054199 TI - Are diagnostic testing and admission rates higher in non-English-speaking versus English-speaking patients in the emergency department? AB - STUDY OBJECTIVE: To determine whether non-English-speaking patients who present to an emergency department have more diagnostic tests ordered, higher admission rate, and longer length of stay in the ED than English-speaking patients for 2 common complaints, chest pain and abdominal pain. METHODS: This prospective, comparative, observational study was conducted at a public hospital ED. The study group was composed of 324 patients (172 non-English-speaking and 152 English speaking) presenting with nontraumatic abdominal pain (148) or chest pain (176). The main outcome measures were admission rates, length of stay in the ED, and diagnostic test and procedure ordering. RESULTS: The mean age for the total sample was 45.8+/-15.5 years (range 14 to 87 years); 45.4% (147/324) of the patients were male. For the non-English-speaking patients, the language distribution was Spanish (31.0%), other (9.0%), Cantonese (5.9%), Hindi (2.5%), Mien (1.5%), Arabic (1.9%), Russian (0.9%), Mandarin (0.6%), and Korean (0.3%). The admission rate was 37.8% for English-speaking patients versus 42.8% for non English-speaking patients in the total sample (mean difference in proportions 5%, 95% confidence interval [CI] -6% to 16%; 34.2% for English-speaking versus 9.1% for non-English-speaking patients presenting with abdominal pain, mean difference in proportions 5%, 95% CI -11% to 21%) and 40.9% for English-speaking versus 45.8% for non-English-speaking patients presenting with chest pain (mean difference in proportions 5%, 95% CI -10% to 20%). Power was 80% to detect a 15% difference in admission rates at an alpha value of.05. There was no statistically significant difference in ordering of diagnostic tests between the non-English speaking and English-speaking patients with chest pain. Non-English-speaking patients with abdominal pain had 5 tests ordered more often than English-speaking patients. The mean difference in proportions (with 95% CIs) for these tests were CBC count 18.4% (5.1% to 31.7%), serum electrolytes 17.9% (3.8% to 31. 9%), urinalysis 20.0% (4.5% to 35.6%), ECG 23.4% (8.6% to 38.2%), and abdominal computed tomographic scan 10.9% (1.0% to 20.8%). There was no statistically significant difference between English-speaking and non-English-speaking patients for ED length of stay in the total sample (mean difference 29.8, 95% CI -37.5 to 97.1 minutes; for the abdominal pain subgroup, mean difference 19.5, 95% CI -74.6 to 113.5 minutes; and for the chest pain subgroup, mean difference 37.9, 95% CI 58.0 to 133.8 minutes). CONCLUSION: Significantly more tests are ordered for non English-speaking patients with abdominal pain in the ED, including 3 times as many abdominal computed tomographic scans. There is no increase in test ordering with non-English-speaking patients with complaints of chest pain in the ED. When comparing English-speaking and non-English-speaking patients, there were no statistically significant differences in admission rates or length of stay in the ED. PMID- 11054200 TI - Improved outcomes in patients with acute allergic syndromes who are treated with combined H1 and H2 antagonists. AB - STUDY OBJECTIVE: Although the addition of H(2) blockers to H(1) antagonists has been promoted for use in anaphylaxis, there have been no large studies establishing the advantage of this approach in treating acute allergic syndromes. In this study we tested the hypothesis that combined H(1) and H(2) blockage results in improved outcomes in patients treated for acute allergic syndromes compared with treatment with H(1) blockade alone. METHODS: In a randomized, double-blind, placebo-controlled trial, 91 adult patients with acute allergic syndromes were treated with either 50 mg of diphenhydramine and saline solution (control group) or with 50 mg of diphenhydramine and 50 mg of ranitidine (active group). These patients were treated with parenteral administration. Patients were recruited from an emergency department at an urban academic medical center. The primary endpoints were resolution of urticaria, angioedema, or erythema at 2 hours after protocol treatment. Areas of cutaneous involvement, heart rates, blood pressures, respiratory findings, and symptom scores were also assessed at baseline, 1 hour, and 2 hours. RESULTS: There were significantly more patients without urticaria at 2 hours among the patients in the active group compared with those in the control group. Both groups had similar proportions of urticaria at baseline. Logistic regression models to predict resolution of urticaria, which accounted for baseline urticarial involvement, showed odds ratios in favor of the active group treatment. Similar findings were observed when the absence of both urticaria and angioedema was considered as the dependent variable. There was not a significant difference between the 2 groups with regard to the absence of erythema or angioedema (irrespective of the presence of urticaria) at 2 hours. Blood pressure and symptoms did not show differences between the 2 groups over time. Lower heart rates were observed 1 hour after treatment in the active treatment group (mean reduction 10 beats/min) compared with those found in the placebo group (mean reduction 6 beats/min). CONCLUSION: These data show that adding H(2) blockers to H(1) antagonists results in additional improvement of certain cutaneous outcomes for patients presenting with acute allergic syndromes. These findings favor the recommendation for using combined H(1) and H(2) antihistamines in acute allergic syndromes. PMID- 11054201 TI - Acute cardiac ischemia in patients with cocaine-associated complaints: results of a multicenter trial. AB - STUDY OBJECTIVE: To describe the characteristics of a large group of patients who presented to emergency departments with cocaine-associated symptoms consistent with acute cardiac ischemia (ACI) and to determine the incidence of confirmed ACI including acute myocardial infarction (AMI) in this population. METHODS: We performed a substudy on all patients in a multicenter prospective clinical trial (the Acute Cardiac Ischemia-Time Insensitive Predictive Instrument [ACI-TIPI] Clinical Trial) that enrolled ED patients with chest pain or other symptoms consistent with ACI including subjects with identified cocaine use. Demographic and clinical features, including initial and follow-up clinical data, ECGs, and tests to determine serum creatine kinase isoenzyme MB subunit concentrations, were analyzed. Diagnoses of AMI followed the World Health Organization criteria for AMI and of angina pectoris, the Canadian Cardiovascular Society Classification. RESULTS: Of the 10,689 patients enrolled in the trial, 293 (2.7%) had cocaine-associated complaints. Among the 10 participating hospitals, the incidence of patients with cocaine-associated symptoms varied from 0.3% to 8.4%. Only 6 patients (2.0%, 95% confidence interval [CI] 0.76% to 4.4%) had a diagnosis of ACI; 4 (1.4%, 95% CI 0.37% to 3.5%) had unstable angina, and 2 (0.7%, 95% CI 0.08% to 2.4%) had AMI. Although patients with cocaine-induced complaints were as likely to be admitted to the coronary care unit compared with all study patients without cocaine use (14% versus 18%, P =.14, difference not significant), these patients were much less likely to have confirmed unstable angina (1.4% versus 9.3%, P <.001) or AMI (0. 7% versus 8.6%, P <.001). Compared with patients younger than 45 years, patients with cocaine usage were more likely to be admitted to the ICU (14% versus 8.0%, P =.0018) but less likely to have confirmed AMI (0.7% versus 2.8%, P =.033). CONCLUSION: Patients presenting to EDs with cocaine-associated chest pain or related symptoms infrequently had ACI, and even less so, AMI. This suggests the need for selectivity in the hospitalization of patients with such cocaine-associated symptoms. PMID- 11054202 TI - Inhaled corticosteroids for acute asthma after emergency department discharge. PMID- 11054203 TI - The semantics of ketamine. PMID- 11054204 TI - Molecular mechanisms of ischemic neuronal injury. AB - Brain ischemia triggers a complex cascade of molecular events that unfolds over hours to days. Identified mechanisms of postischemic neuronal injury include altered Ca(2+) homeostasis, free radical formation, mitochondrial dysfunction, protease activation, altered gene expression, and inflammation. Although many of these events are well characterized, our understanding of how they are integrated into the causal pathways of postischemic neuronal death remains incomplete. The primary goal of this review is to provide an overview of molecular injury mechanisms currently believed to be involved in postischemic neuronal death specifically highlighting their time course and potential interactions. PMID- 11054205 TI - Evidence-based emergency medicine/clinical question. How good is a negative cranial computed tomographic scan result in excluding subarachnoid hemorrhage? PMID- 11054206 TI - Evidence-based emergency medicine/perspectives. One is the loneliest number: be skeptical of evidence summaries based on limited literature reviews. PMID- 11054207 TI - Evidence-based emergency medicine/perspectives. One is the only number that you'll ever need! PMID- 11054208 TI - Massive sertraline overdose. AB - Selective serotonin reuptake inhibitor medications are considered relatively safe even in overdose. We report a massive overdose of sertraline with the highest serum sertraline concentration reported to date. Clinical features of this patient were confusion, agitation, myoclonus, hyperreflexia, fever, and creatine kinase elevation. This case may represent serotonin syndrome caused by sertraline overdose alone. PMID- 11054209 TI - Update on emerging infections from the centers for disease control and prevention. PMID- 11054210 TI - A guide to your first employment contract. PMID- 11054211 TI - Important announcement. PMID- 11054212 TI - A farewell to arm. PMID- 11054213 TI - Presidential address: entering the third millennium. PMID- 11054214 TI - Surgical treatment of venous malformations in Klippel-Trenaunay syndrome. AB - PURPOSE: Klippel-Trenaunay syndrome (KTS) is a complex congenital anomaly, characterized by varicosities and venous malformations (VMs) of one or more limbs, port-wine stains, and soft tissue and bone hypertrophy. Venous drainage is frequently abnormal because of embryonic veins, agenesis, hypoplasia, valvular incompetence, or aneurysms of deep veins. We previously reported on the surgical management of KTS. In this article, we update our experience. METHODS: Twenty patients with KTS underwent surgical treatment for VMs between July 1, 1987, and January 1, 2000. This group represented 6.9% of 290 patients with KTS who were seen at our institution during this 12.5-year study period. Surgical indications, venous anatomy (determined with duplex scan, contrast phlebography, magnetic resonance imaging or magnetic resonance phlebography), operative procedures, and complications were reviewed, and outcomes were recorded. RESULTS: Twelve male and eight female patients (mean age, 23.4 years; range, 7.7-40.6 years) underwent 30 vascular surgical procedures in 21 lower limbs. All 20 patients (100%) had varicose veins or VMs, 13 (65%) had port-wine stains, and 18 (90%) had limb hypertrophy. Pain was the most common complaint, which was present in 16 patients (80%), followed by swelling in 15 (75%), bleeding in 8 (40%), and superficial thrombophlebitis and cellulitis in 3 (15%). Imaging confirmed patent deep veins in 18 patients, hypoplastic femoral vein in 1, and entrapped popliteal veins bilaterally in 1. Four patients (20%) had large persistent sciatic veins (PSVs). The CEAP clinical classification was C-3 for 17 patients (85%), C-4 for 1 patient (5%), and C-6 for 2 patients (10%). Stripping of large lateral veins, avulsion, and excision of varicosities or VMs were performed on all limbs. Three patients required staged resections. The release of entrapped popliteal veins was performed in both limbs of one patient; another underwent a popliteal-saphenous bypass graft. One patient underwent excision of a PSV. Open and endoscopic perforator vein ligation was performed in one patient each. Two patients (12%) had hematomas that required evacuation. No patients had caval filter placement; none had postoperative deep venous thrombosis or pulmonary embolus. The mean follow-up was 63.6 months (range, 0-138 months). All patients reported initial improvement, but some varicosities recurred in 10 patients (50%), an ulcer did not heal in one, and a new ulcer developed in one, 8 years after surgery. Three patients underwent reoperation for recurrent varicosities. Follow-up CEAP scores were C-2 in 10 patients (50%), C-3 in 6 patients (30%), C-4 and C-5 in 1 patient each (5%), and C-6 in 2 patients (10%). Clinical scores improved from 4.3 +/- 2.2 to 3.1 +/- 2.3. (P =.03). CONCLUSIONS: The management of patients with KTS continues to be primarily nonoperative, but those patients with patent deep veins can be considered for excision of symptomatic varicose veins and VMs. Although the recurrence rate is high, clinical improvement is significant, and reoperations can be performed if needed. Occasionally, deep vein reconstruction, excision of PSVs, or subfascial endoscopic perforator surgery is indicated. Because KTS is rare, patients should receive multidisciplinary care in qualified vascular centers. PMID- 11054215 TI - Free tissue transfer provides durable treatment for large nonhealing venous ulcers. AB - BACKGROUND: Most venous ulcers (VUs) will heal with conventional treatment, which focuses on improving regional venous hemodynamics. This treatment, however, often fails to heal large, recurrent VUs that are associated with severe lipodermatosclerosis (LDS). These complicated ulcers may require correction of local venous hemodynamics and replacement of the surrounding LDS with healthy tissue. We report our experience managing 24 especially difficult VUs with debridement and free flap coverage. PATIENTS AND METHODS: Between 1987 and 1997, 25 free flap procedures were performed in 22 patients for 24 recalcitrant VUs. Ulcers had been present for a mean of 5.24 years and had failed to heal with conservative therapy and split-thickness skin grafts (STSGs) (mean, 2.2). Eleven patients (46%) had exposed bone, tendon, or joint. At operation the area of LDS was excised, and all perforating veins were ablated. The defects after excision ranged from 100 to 600 cm(2) (mean, 237 cm(2)). The free flap was inset within the defect and covered with an STSG. RESULTS: We healed all 24 ulcers with free tissue transfer (one patient required a second flap after the first failed). There were no deaths. Local complications that required repeat STSG occurred in three (13%) of the 24 successful flap transfers. Four other flaps had minor local complications that healed with local wound care. Follow-up was available for 21 of the 24 successful flap transfers. No recurrent ulcers were identified in the territory of the flap after a mean of 58 months, but three patients had new ulcers in the same leg after 6 to 77 months. Patients with severe complications were hospitalized longer than those with minor or no complications (45.7 vs 12.8 days, P <.01), and their hospital charges were greater ($76,681 vs $30,428, P <.01). CONCLUSION: Free tissue transfer can provide rapid healing and long-term relief from severe VUs that are unable to be treated with conventional therapy. This technique improves venous hemodynamics, removes all liposclerotic tissue, provides an abundant blood supply, and resolves the tissue-related components of chronic ulceration. Although further work is needed to determine the optimal indications, this technique provides a durable treatment for especially recalcitrant ulcers. PMID- 11054216 TI - Practicability and quality of outpatient management of acute deep venous thrombosis. AB - PURPOSE: The purpose of this study was to review the practicability and quality of a standardized management approach of deep venous thrombosis (DVT) provided by private practices. METHODS: There were 152 consecutive patients and 156 episodes. We determined the patients' diagnoses with estimation of clinical probability, D dimers, duplex ultrasound scan, and venography. Patients were treated on an outpatient basis on principle, with dalteparin, phenprocoumon, different modalities of external leg compression, and deliberate ambulation. We followed up at 4 weeks. RESULTS: Proximal DVT was diagnosed in 101 episodes (65%). Results of the D-dimer test were false-negative in 6%, and venography was indicated in 15%. Calf vein thrombosis was found in 55 patients. Results of the D-dimer test were false-negative in 30%, and venography was required in 37%. Eleven patients were hospitalized (9 for thrombectomy or thrombolysis), and 145 episodes (93%) were treated according to our standardized approach (provided by the referring physicians alone in 43%). For 5 days, dalteparin was injected by the patients themselves or their relatives, in 78% of the cases. The international normalized ratio values were more than 2 in 88% of the cases, with no difference between providers. In all but two cases, external leg compression was applied immediately: a modified Unna's boot in 28% and compressing stockings in 72% (Sigvaris 503 in 91%; calf length in 100% of distal DVT, and 83% of proximal DVT). During follow-up, there was no clinical evidence of recurrence or progression, 1 possible pulmonary embolism, 1 injection site hematoma, and 1 hospitalization unrelated to the DVT. CONCLUSION: Diagnosis of proximal DVT is straightforward, but calf DVT often requires venographic confirmation because of discrepancies among clinical probability, D-dimer estimation, and ultrasound scan. Outpatient treatment can be offered to patients who can comply with the regimen. The quality of anticoagulation is in accordance with published data, and compliance with external leg compression is almost perfect. PMID- 11054217 TI - Compression and walking versus bed rest in the treatment of proximal deep venous thrombosis with low molecular weight heparin. AB - OBJECTIVE: The purpose of this randomized controlled trial was to evaluate the benefits of compression and walking exercises in comparison with bed rest in the acute stage of proximal deep venous thrombosis (DVT). METHODS: Forty-five patients with proximal DVT that was proved with compression ultrasound scan or phlebography were randomized into three groups. Group A consisted of 15 patients who received inelastic compression bandages (Unna boots on the lower leg, adhesive bandages on the thigh), and group B consisted of 15 patients who received thigh-length compression stockings, class II. Group C consisted of 15 patients who underwent bed rest and no compression. All patients received dalteparin, 200 IU/kg per body weight, subcutaneously every 24 hours. The clinical characteristics of the three groups were comparable. Primary end points were the reduction of pain assessed daily with the Visual Analogue Scale and the Lowenberg test, the reduction of leg circumference at the ankle and calf levels, and the improvement of clinical scores. The daily walking distance was measured with a pedometer. Safety parameters were ventilation-perfusion scans and duplex ultrasound scans performed on days 0 and 9. RESULTS: The daily walking distance was between 600 and 12,000 m in the compression groups and averaged 66 m in the bed rest group. The pain level showed a statistically significant reduction starting after the second day in the compression groups (A and B) and after 9 days in the bed rest group C (P <.05). The same was true for the measurement of leg circumference. Improvement of the clinical scores was significantly better in the compression groups compared with the bed rest group (P <.01). There was no significant difference concerning the occurrence of new pulmonary emboli and regression of thrombus diameter. Progression of thrombi in the femoral vein was greater and occurred more frequently in the bed rest group than in the other two groups (P = not significant). CONCLUSION: Mobile patients with acute proximal DVT treated with low molecular weight heparin should be encouraged to walk with compression bandages or medical compression stockings. The rate of resolution of pain and swelling is significantly faster when the patient ambulates with compression. The risk of pulmonary embolism is not significantly increased by this approach. PMID- 11054218 TI - Markers of plasma coagulation and fibrinolysis after acute deep venous thrombosis. AB - OBJECTIVE: Plasma markers of coagulation and fibrinolysis have proved sensitive in the initial diagnosis of acute deep venous thrombosis (DVT). The purpose of this study was to examine the evolution and utility of measuring D-dimer and prothrombin fragment 1+2 (F 1+2) levels after an acute DVT. METHODS: Subjects with DVT confirmed by ultrasonography had quantitative plasma D-dimer and F 1+2 levels determined before anticoagulation. Ultrasound scan and coagulation studies were repeated at 3, 7, and 14 days; 1 month; and every 3 months for 1 year. RESULTS: Sixty-one patients with a median initial thrombus score of 3 (interquartile range, 2-7) were followed up for 266 days (interquartile range, 91.5-364 days). Initial D-dimer levels were elevated in 92.7% of patients and were associated with thrombus extent (P =.003), whereas F 1+2 levels were increased in 94.5% of patients and were lower in patients with isolated calf vein thrombosis (P =.001). Initial D-dimer (P =.002) and F 1+2 levels (P =.009) were significantly higher in the 26 (43%) patients with recurrent thrombosis during follow-up. Initial D-dimer levels of 2000 ng/mL or greater were predictive of recurrent events after both proximal and isolated calf vein thrombosis. Although interval increases in these markers had little value in detecting recurrent thrombotic events, D-dimer levels of 1000 ng/mL or greater and 500 ng/mL or greater had respective sensitivities of 89.3% and 100% in detecting early and late recurrences. Corresponding specificities were 35.6% and 53.9%. CONCLUSIONS: Initial D-dimer levels are determined by total thrombus load and remain elevated long after an acute DVT. F 1+2 levels are less sensitive to thrombus score and return to baseline more quickly. Initial levels of these markers may have some utility in predicting the risk of ultrasound scan-documented recurrences, whereas increased D-dimer levels are a sensitive but nonspecific marker of these events. PMID- 11054219 TI - Lessons learned from a 6-year clinical experience with superior vena cava Greenfield filters. AB - PURPOSE: Therapy to prevent pulmonary embolism (PE) resulting from upper extremity deep venous thrombosis (UEDVT) remains controversial despite an increasing incidence of DVT of upper extremity origin. The purpose of this study was to evaluate the results of 72 superior vena cava Greenfield filters (SVC-GFs) placed in patients at risk for PE arising from UEDVT. METHODS: During the past 78 months, we placed SVC-GFs in 72 patients with UEDVT in whom anticoagulation was either deemed contraindicated (n = 67) or proved ineffective in preventing recurrent PE (n = 4) or extension of the thrombus (n = 1). There were 25 male (35%) and 47 (65%) female patients whose ages ranged from 25 to 99 years (mean, 74 years). Follow-up ranged from 10 days to 78 months (mean, 7.8 months). Sequential chest radiographs revealed no filter migration or displacement in 26 patients. RESULTS: Thirty-four patients died in the hospital of causes unrelated to the SVC filter or recurrent thromboembolism (mean time to death, 20 days). Follow-up of the surviving 38 patients ranged from 1 month to 78 months (mean, 22 months); none of these patients were seen with any evidence of PE. One SVC-GF was incorrectly discharged into the innominate vein and left in place. This vein remains patent 2 months after insertion without evidence of filter migration. CONCLUSIONS: We think that insertion of SVC-GFs is a safe, efficacious, and feasible therapy and may prevent recurrent thromboembolism in patients with UEDVT who are resistant to anticoagulation or have contraindications to anticoagulation. PMID- 11054220 TI - The percutaneous greenfield filter: outcomes and practice patterns. AB - OBJECTIVE: The percutaneous steel Greenfield filter (PSGF) is similar in appearance to the titanium Greenfield filter (TGF) but differs in the length and orientation of the attachment hooks and in the over-the-wire delivery system. Because these differences improve ease of insertion and attachment, they may affect patient outcomes and physician practices. The purpose of this study was to evaluate the performance of the PSGF relative to the TGF and to determine whether there had been a change in physician practices. METHODS: The Michigan Filter Registry contains data for a prospective cohort of 2188 patients with Greenfield filters. Procedural and long-term outcomes for patients with a PSGF were abstracted. These events were compared with rates for Registry patients who had a TGF. Trends for indication for placement, delivery route, and filter location were also compared with published series. RESULTS: Since 1995, 600 PSGFs have been placed in 599 patients. A 1-year mortality rate of 42% left 349 patients available for annual follow-up, and studies were completed for 231 (66%). Periprocedural events occurred in 2.5% of cases with associated morbidity in 1.5%. The rate of new pulmonary embolism was 2.6%, and vena caval patency was 98.3%. The combined rate of new venous thromboembolic events was 12.5%. Left sided femoral vein placements increased to 20%, and the major indication for filter placement has become prophylaxis (46%). CONCLUSIONS: The PSGF is similar to the TGF with respect to patient outcomes, and it provides decreased rates of asymmetry along with excellent fixation. The flexible carrier system has allowed more frequent access through the left femoral vein. The ease of use and favorable patient outcomes have resulted in more frequent placement for prophylactic indications. PMID- 11054221 TI - Differences in pressures of the popliteal, long saphenous, and dorsal foot veins. AB - PURPOSE: The purpose of this study was to examine the relationship among pressures obtained simultaneously in the popliteal, long saphenous, and dorsal foot veins. METHOD: Eight limbs were studied. One limb had an isolated popliteal vein reflux, and two had moderate long saphenous vein incompetence. No perforator or short saphenous vein insufficiency was detected. Pressures and recovery times of the popliteal/tibial and long saphenous veins were obtained with cannulation at the ankle level and insertion of catheters with a pressure transducer tip. The dorsal foot vein pressure was measured with the insertion of a scalp needle (14 gauge) connected to an external transducer. During 10 toe stands, recordings were simultaneously made in the three veins at the level of the knee joint, in the middle third of the calf, and 5 to 7 cm above the ankle with all the transducers at the same level (ie, same reference point). RESULTS: In one limb the popliteal/tibial pressure increased at all calf levels, whereas pressures decreased in both saphenous and dorsal foot veins. The pressures decreased in all three systems in the remaining seven limbs. There was no statistical difference between the pressure drop in the long saphenous vein and the deep vein. However, the decrease of the dorsal foot venous pressure was significantly more marked compared with the other two veins at all levels. The recovery time was significantly increased in the long saphenous vein compared with the deep vein; recovery time was further prolonged in the dorsal foot vein. CONCLUSION: The dorsal foot, long saphenous, and popliteal/posterior tibial veins clearly exhibit different pressure waveforms in response to calf exercise. The postexercise pressure, the percentage pressure drop, and the recovery times are widely different, which indicates that the three veins behave hydraulically as separate compartments in limbs without significant venous insufficiency. PMID- 11054222 TI - Management of symptomatic and asymptomatic popliteal venous aneurysms: a retrospective analysis of 25 patients and review of the literature. AB - OBJECTIVE: Popliteal venous aneurysms (PVAs) are an uncommon but potentially life threatening disease because they can be a source for pulmonary emboli (PE). With the widespread use of venous duplex scanning, PVAs are increasingly found in patients with deep or superficial vein insufficiency, and questions have arisen as to the management of these aneurysms. The purpose of this study was to review our experience in the management of PVAs diagnosed in patients with PE and in patients with chronic venous diseases. METHODS: Twenty-five patients with PVAs were treated in two centers between 1985 and 1999. There were 20 women and five men, with an average age ranging from 33 to 79 years (mean age, 59 years). Twenty four percent (6/25) presented with PE, and 76% (19/25) of PVAs were discovered during investigation for chronic venous disease (varicosities, n = 13; post thrombotic symptoms, n = 6). The diagnosis of PVA was achieved in all cases with venous duplex scanning and phlebography. Aneurysms were located in the proximal popliteal vein (n = 17) and at the saphenopopliteal junction (n = 8). Seventy-two percent (18/25) of PVAs were saccular, and 40% (10/25) had an intraluminal thrombus. Two patients with PE underwent cardiac arrest, with one requiring a pulmonary embolectomy. The Fisher exact test showed a statistically significant correlation between PE and the presence of thrombus (50% vs 7% without thrombus, P =.02). Aneurysms were treated with tangential aneurysmectomy and lateral venorrhaphy (n = 19), resection with end-to-end anastomosis (n = 2), resection with interposition of the greater saphenous vein (n = 2) or the superficial femoral vein (n = 1), and resection with vein transposition (n = 1). Two patients who experienced a PE had an inferior vena cava filter placement before surgical repair of the PVA. RESULTS: Mean follow-up was 63 months (range, 11-168 months). No operative deaths occurred, and no patient had evidence of a recurrent PE. Postoperative minor complications (20%) included transient common peroneal nerve palsy (n = 2), postoperative hematoma (n = 2), and wound infection (n = 1). Postoperative thrombosis of the surgical repair occurred in three patients, and patency was restored with anticoagulation therapy. CONCLUSION: Despite its rarity, PVAs should be ruled out with venous duplex scanning in patients with PE and in patients presenting with chronic venous diseases. Because of the unpredictable risk of thromboembolic complications, surgical treatment that is accompanied by a low morbidity rate is indicated in all PVAs. Tangential aneurysmectomy with lateral venorrhaphy is the procedure of choice. PMID- 11054223 TI - Target selection for surgical intervention in severe chronic venous insufficiency: comparison of duplex scanning and phlebography. AB - PURPOSE: The goal of this study was to determine whether duplex scanning (DS) alone, compared with ascending phlebography (AP) and descending phlebography (DP), would have been sufficient to guide treatment of severe chronic venous insufficiency (CVI), CEAP Clinical Classes 5 and 6. METHODS: Beginning in 1994, patients presenting to the VA Sierra Nevada Vascular Clinic with ulceration due to CVI, CEAP Clinical Classes 5 and 6, were examined with DS, AP, and DP. Phlebography mainly guided surgical interventions. The ability of DS findings to select surgical interventions, with the aims of diversion of reflux from area of trophic skin or reduction of global venous hypertension was compared with phlebography. Of the 33 male patients (age, 29-70 years; average, 55 years) considered for operative interventions between January 1994 and November 1999, 30 were selected for operative treatment. RESULTS: DS was 100% sensitive and specific for detection of complete occlusion of the superficial femoral vein (10/10) and for saphenous incompetence; sensitivity was 95% (19/20); and specificity was 100%. However, DS failed to reveal subtle changes in recanalized femoral veins because of prior thrombophlebitis, which was uncovered by AP in six of 23 patent femoral veins. There were 16 positive findings on AP of residual thrombophlebitis, of which six were not read on DS. Sensitivity was 63%, specificity was 100%, the positive predictive value was 100%, and the negative predictive value was 53%. Reflux grading with DP agreed with DS in 23 of 33 cases or varied by one grade in five of 33 cases: sensitivity, 82%; specificity, 75%; positive predictive value, 96%; and negative predictive value, 37%. Kistner grade 4 reflux involving the superficial femoral and popliteal veins was noted by DP in five of the 33 cases when DS described reflux as "moderate." Incompetent superficial femoral vein valve stations in the upper third of the vein, which caused primary reflux, were clearly defined by DP in four of 33 cases; valve location was not well defined by DS. Below-knee perforator identification with DS was difficult; this was related to the severity of lipodermatosclerosis and the presence of ulceration. The number of perforators described at operation with subfascial endoscopic perforator surgery (n = 13) averaged 6 +/- 2, whereas AP identified an average of 4 +/- 2 in supramalleolar area. In four men, two previously undiagnosed caval and two iliac obstructions were detected with AP; one was corrected with Palma bypass grafting. Follow-up at 4 to 60 months (average, 40 months) showed four ulcer recurrences among 30 patients who were operated on. Two patients underwent repeat operations on the basis of repeated phlebographic study and are cured at this time, one patient was healed with conservative therapy, and one patient is lost to follow-up. CONCLUSIONS: DS would have been inadequate for identifying surgical targets in CVI, CEAP Clinical Classes 5 and 6. DS overlooked iliac and caval lesions. Potential valveplasty sites, which were only delineated on DP, resulted in four valveplasties in the upper third of the superficial femoral vein for grade 4 reflux. AP localized mid- to upper-leg perforators, but neither AP nor DP detected perforators in the range of 5 to 10 cm above the calcaneus. The net effect of phlebography was a choice for deep interventions in five (17%) of 30 cases, which would not have been possible with DS alone. The identification of iliocaval occlusion influenced the decision, based on prior experience, not to perform distal procedures in three cases. PMID- 11054224 TI - The relationship between lower limb symptoms and superficial and deep venous reflux on duplex ultrasonography: The Edinburgh Vein Study. AB - BACKGROUND: Previous work from this group has demonstrated the relationships between lower limb symptoms and the presence and severity of trunk varicose veins as seen on clinical examination to be generally weak, symptom specific, and gender dependent. OBJECTIVE: This study was undertaken to investigate the relationships in the general population between lower limb symptoms and the presence of superficial or deep venous reflux. METHODS: A cross-sectional study was made of an age-stratified random sample of 1566 subjects (699 men and 867 women) aged 18 to 64 selected from 12 general practices in Edinburgh, Scotland. Subjects completed a self-administered questionnaire regarding symptoms (heaviness/tension, a feeling of swelling, aching, restless legs, cramps, itching, tingling) and underwent duplex ultrasound examination of the superficial and deep venous systems of both legs. Reflux of 0.5 seconds or greater was considered pathologic. Deep venous reflux was defined as reflux in at least the popliteal vein. RESULTS: There was a significant positive relationship between isolated superficial reflux and the presence of heaviness/tension (P <.025, both legs) and itching (P =.002, left leg) in women. Isolated superficial reflux in men was not significantly positively associated with any symptom. Isolated deep venous reflux was not significantly related to any symptom in either leg in either sex. Combined reflux was related to a feeling of swelling (P =.018, right leg; P =.0022, left leg), cramps (P =.0049, left leg) and itching (P =.0043, left leg) in men, and aching (P =.03, right leg) and cramps (P =.026, left leg) in women. CONCLUSION: In the general population, only certain lower limb symptoms were related to the presence of reflux on duplex ultrasound scanning. The strongest relationships were observed in the left legs of men with combined superficial and deep reflux. PMID- 11054225 TI - The efficacy of the new SCD response compression system in the prevention of venous stasis. AB - OBJECTIVE: The current commercially available sequential intermittent pneumatic compression device used for the prevention of deep venous thrombosis has a constant cycle of 11 seconds' compression and 60 seconds' deflation. This deflation period ensures that the veins are filled before the subsequent cycle begins. It has been suggested that in some positions (eg, semirecumbent or sitting) and with different patients (eg, those with venous reflux), refilling of the veins may occur much earlier than 60 seconds, and thus a more frequent cycle may be more effective in expelling blood proximally. The aim of the study was to test the effectiveness of a new sequential compression system (the SCD Response Compression System), which has the ability to detect the change in the venous volume and to respond by initiating the subsequent cycle when the veins are substantially full. METHODS: In an open controlled trial at an academic vascular laboratory, the SCD Response Compression System was tested against the existing SCD Sequel Compression System in 12 healthy volunteers who were in supine, semirecumbent, and sitting positions. The refilling time sensed by the device was compared with that determined from recordings of femoral vein flow velocity by the use of duplex ultrasound scan. The total volume of blood expelled per hour during compression was compared with that produced by the existing SCD system in the same volunteers and positions. RESULTS: The refilling time determined automatically by the SCD Response Compression System varied from 24 to 60 seconds in the subjects tested, demonstrating individual patient variation. The refilling time (mean +/- SD) in the sitting position was 40.6 +/- 10. 0 seconds, which was significantly longer (P <.001) than that measured in the supine and semirecumbent positions, 33.8 +/- 4.1 and 35.6 +/- 4.9 seconds, respectively. There was a linear relationship between the duplex scan-derived refill time (mean of 6 readings per leg) and the SCD Response device-derived refill time (r = 0.85, P <. 001). The total volume of blood (mean +/- SD) expelled per hour by the existing SCD Sequel device in the supine, semirecumbent, and sitting positions was 2.23 +/ 0.90 L/h, 2.47 +/- 0.86 L/h, and 3.28 +/- 1.24 L/h, respectively. The SCD Response device increased the volume expelled to 3.92 +/- 1.60 L/h or a 76% increase (P =.001) in the supine position, to 3.93 +/- 1.55 L/h or a 59% increase (P =. 001) in the semirecumbent position, and to 3.97 +/- 1.42 L/h or a 21% increase (P =.026) in the sitting position. CONCLUSIONS: By achieving more appropriately timed compression cycles over time, the new SCD Response System is effective in preventing venous stasis by means of a new method that improves on the clinically documented effectiveness of the existing SCD system. Further studies testing its potential for improved efficacy in preventing deep venous thrombosis are justified. PMID- 11054226 TI - Defining the role of extended saphenofemoral junction ligation: a prospective comparative study. AB - OBJECTIVE: This study explores the added effect of extended saphenofemoral junction (SFJ) ligation when the greater saphenous vein (GSV) has been eliminated from participating in thigh reflux by means of endovenous obliteration. GSV obliteration, unlike surgical stripping, can be done with or without SFJ ligation to isolate and study SFJ ligation's specific contribution to treatment results. METHODS: Sixty limbs treated with SFJ ligation and 120 limbs treated without high ligation were selected from an ongoing, multicenter, endovenous obliteration trial on the basis of their having primary varicose veins, GSV reflux, and early treatment dates. RESULTS: Five (8%) high-ligation limbs and seven (6%) limbs without high ligation with patent veins at 6 weeks or less were excluded as unsuccessful obliterations. Treatment significantly reduced symptoms and CEAP clinical class in both groups (P =.0001). Recurrent reflux developed in one (2%) of 49 high-ligation limbs and eight (8%) of 97 limbs without high ligation by 6 months (P =.273). New instances of reflux did not appear thereafter in 57 limbs followed to 12 months. Recurrent varicose veins occurred in three high-ligation limbs and four limbs without high ligation by 6 months and in one additional high ligation limb and two additional limbs without high ligation by 12 months. Actuarial recurrence curves were not statistically different with or without SFJ ligation (P >.156), predicting greater than 90% freedom from recurrent reflux and varicosities at 1 year for both groups. CONCLUSION: These early results suggest that extended SFJ ligation may add little to effective GSV obliteration, but our findings are not sufficiently robust to warrant abandonment of SFJ ligation as currently practiced in the management of primary varicose veins associated with GSV vein reflux. PMID- 11054227 TI - The impact of isolated lesser saphenous vein system incompetence on clinical signs and symptoms of chronic venous disease. AB - PURPOSE: The purpose of this study was to determine the patterns of isolated lesser saphenous vein (LSV) system incompetence and correlate the distribution and extent of such reflux with symptoms and signs of chronic venous disease (CVD). METHODS: During a 3-year period, 2254 limbs in 1682 patients with signs and symptoms of CVD were evaluated with color flow duplex scanning. Extremities with isolated reflux in the LSV system were selected for this study. Limbs with perforating venous reflux connected to this system only were also included. Limbs that had marked reflux in the greater saphenous or deep vein, that had a documented history of deep venous thrombosis, and that previously underwent surgery or sclerotherapy were excluded. The clinical severity of the limbs was graded with the CEAP classification system. RESULTS: There were 226 limbs in 200 patients with reflux in the LSV system; 61% were female patients with a mean age of 49 years (range, 18-82 years). There were 174 patients (87%) with unilateral and 26 with bilateral disease, and 41% of the limbs belonged in CVD class 2, 26% in class 3, 12% in class 4, 3.5% in class 5, and 3% in class 6. Classes 0 and 1 were present in 14.5% of the limbs. Symptoms were present in 139 limbs (61.5%). Some degree of ache or burning sensation was the most frequent symptom (41%), followed by itching (32%), heaviness (29%), cramps (24%), and restless limbs (18%). Reflux in the main trunk of the LSV was the most prevalent (177 limbs [78%]), followed by the saphenopopliteal junction (146 limbs [64.6%]), the vein of Giacomini (39 limbs [17%]) and the gastrocnemial vein (23 limbs [10%]). Reflux involving both the saphenopopliteal junction and the LSV was seen in 50% of limbs, but almost any other combination of reflux was present, which indicated the complexity of this system. Perforator vein incompetence was detected in 56 limbs (25%). We found 83 perforator veins, resulting in a mean of 1.5 veins per limb. Both the number of incompetent perforator veins and the extent of superficial reflux correlated with clinical severity. Four main types of termination of the LSV were identified with at least nine variations. The LSV was duplicated for at least half of its length in five limbs (2.2%). Nonsaphenous reflux was detected in seven limbs (3.1%). Superficial vein thrombosis in the LSV system was found in eight limbs (3.5%), and in the gastrocnemial vein it was found in four (1.8%). CONCLUSIONS: Isolated LSV system incompetence can cause the entire range of signs and symptoms of CVD. Clinical deterioration is associated with a longer extent of reflux and perforator incompetence. Classes 2 to 4 are the most frequent clinical presentations, whereas classes 5 and 6 are uncommon. The complex anatomy of this system and the great variation in the patterns of reflux warrant the use of color flow duplex scanning before planning treatment. PMID- 11054228 TI - The value of air plethysmography in predicting clinical outcome after surgical treatment of chronic venous insufficiency. AB - PURPOSE: The role of air plethysmography (APG) as a predictor of clinical outcome after surgery in venous disease is yet to be defined. The purpose of this study was to investigate the value of APG in predicting clinical outcome after venous surgery for chronic venous insufficiency (CVI). METHODS: Seventy-three extremities in 71 patients with Class 3 through 6 CVI were assessed preoperatively with CEAP (c linical, e tiologic, a natomic, p athophysiologic) criteria, standing reflux duplex ultrasound scan, and APG with measurements of preoperative venous filling index (VFI), venous volumes, ejection fraction, and residual volume fraction. After surgical treatment of the affected limbs, repeat APG studies were obtained within 6 weeks. Established venous reporting standards were used for follow-up to calculate clinical symptom scores (CSSs) in each patient. RESULTS: Superficial venous reflux occurred alone in 24 limbs or in conjunction with perforator incompetence in 26 limbs. Deep and superficial reflux, with or without perforator incompetence, was found in 16 limbs, and seven limbs had isolated deep insufficiency. Follow-up was available in 60 of 71 patients (mean period, 44.3 months). Postoperative APG demonstrated significant hemodynamic changes after surgery as measured with VFI, venous volumes, ejection fraction, and residual volume fraction. Mean CSSs decreased from 7.35 +/- 0.56 preoperatively to 1.79 +/- 0.32 at late follow-up after surgery (P <.001). With the use of logistic regression, the parameter correlating most closely with clinical outcome was the VFI. A normal postoperative VFI (40 kg/m(2) or > or =35 kg/m(2) with obesity related comorbidity) in whom non-surgical treatment options were unsuccessful. Additional research is needed to examine the long-term benefits of weight loss following bariatric surgery, particularly with respect to obesity-related comorbidities. PMID- 11054243 TI - Morbid obesity: perceptions of character and comorbidities in Falstaff. PMID- 11054244 TI - Open vs. laparoscopic vertical banded gastroplasty: a case control study with a 1 year follow-up. AB - BACKGROUND: Vertical Banded Gastroplasty (VBG) is one of the most common bariatric operations. It can be performed by open or laparoscopic methods. The purpose of this study was to analyze and compare the 1-year results of 40 patients who underwent laparoscopic (20) and open (20). METHODS: The initial 20 patients undergoing Laparoscopic VBG and the initial 20 patients in whom an Open VBG were performed in our Institution were comparatively evaluated. Demography, surgical details, complications, and 1-year weight loss were analyzed. RESULTS: Both groups were highly comparable in terms of age, sex and body mass index. Laparoscopic VBG was a more prolonged procedure (median 4 hr) than the open VBG (median 3 hr). On the other hand, hospital stay was significantly shorter in the laparoscopic procedure (median 10 days for the open and 6 days for the laparoscopic). One year weight loss and complications were similar in both groups. CONCLUSIONS: Laparoscopic VBG is a safe procedure for the treatment of morbid obesity. This initial series shows comparable results. PMID- 11054245 TI - Micropouch gastric bypass: indications for gastrostomy tube placement in the bypassed stomach. AB - BACKGROUND: Since 1994 we have performed the "micropouch" gastric bypass (MGB) procedure for morbid obesity. In our MGB operation, the transected gastric reservoir is limited to the cardia of the stomach. The Roux and biliopancreatic limbs measure 200 and 150 cm, respectively; radiographic or endoscopic access to the bypassed stomach is therefore excluded. METHODS: A retrospective analysis of 1,120 MGB operations revealed that 33 patients (2.9%) developed a potentially life-threatening postoperative complication requiring emergency re-operation or prolonged hospitalization. Complications included anastomotic leaks, fascial dehiscence, gastro-enteric obstructions, peptic ulcers, and pancreatitis. Each complication was analyzed to determine if gastrostomy tube (G-tube) placement at the time of gastric bypass could have affected complication outcome or obviated the need for repeat laparotomy. RESULTS: In 19 patients (1.6% of total cohort), G tube placement would have significantly affected complication outcome. In four instances, it would have prevented emergency re-operation. CONCLUSIONS: Routine gastrostomy tube placement at the time of gastric bypass is not necessary in all patients. For patients who are at high risk for a gastro-enteric obstruction or an anastomotic leak, G-tube placement is recommended and is often therapeutic. PMID- 11054246 TI - Hand-assisted laparoscopic Roux-en-y gastric bypass: aspects of surgical technique and early results. AB - BACKGROUND: The efficacy of Roux-en-y gastric bypass (RYGBP) for morbid obesity is well documented. We investigated the role of the Hand-assisted laparoscopic technique for performing RYGBP. METHODS: In an open series, 13 patients (all female, median age 38, BMI 45 kg/m(2)) underwent Hand-assisted laparoscopic RYGBP. The HandPort was introduced through an 8-cm right subcostal incision. The stomach was always completely transected. The Roux limb was made > 50 cm and brought to the proximal gastric pouch (4 x 3 cm) behind the colon and the excluded stomach. A circular stapler (no. 21) was used for the gastrojejunostomy, with the anvil introduced through a gastrotomy. RESULTS: The HandPort device could be successfully placed and allowed good working conditions in all patients. Median duration of surgery (including learning-curve time) and postoperative hospital stay were 205 min and 5 days, respectively. The amount of morphine needed (PCA) during postoperative day 1-3 were 45, 32 and 18 mg, respectively. One patient (8%) was converted to full laparotomy for safe closure of a small perforation of the proximal gastric pouch caused by the anvil of the circular stapler. All patients made an uneventful recovery. Two patients needed endoscopic dilatation of a relative stricture at the gastrojejunostomy. CONCLUSION: We believe that Hand-assistance makes Lap-RYGBP faster and safer without losing the essential benefits of total laparoscopy. PMID- 11054247 TI - Improvement in co-morbidities following weight loss from gastric bypass surgery. AB - BACKGROUND: Clinical observation reveals a close association between morbid obesity and a variety of serious medical conditions. This report describes the changes observed in some of these co-morbid conditions, following weight loss achieved by silastic ring gastric bypass (SRGBP). METHODS: Between 1990 and 1998, 157 severely obese patients aged 15-62 years underwent SRGBP. Initial and follow up data was recorded prospectively on a computerized database, with minor subsequent additions being achieved by phone call or questionnaire. Particular attention was given to associated co-morbidities and improvement in these that occurred during follow-up. Median pre-operative BMI was 45 (33-97). Patients were followed for a median 2.5 years. At 2 years post-SRGBP, median BMI was 28 (20 52). Weight loss was statistically significant (p<0.0001). RESULTS: Before surgery 42 patients were being treated for hypertension and 34 for asthma. Withdrawal of all medication for these conditions was achieved sometime after surgery in 18 and 17 patients respectively. NIDDM was present in 19 patients before surgery and subsequently resolved completely in 18. Eleven of the 12 patients with recognized obstructive sleep apnea before surgery had resolution of this after surgery. Dyslipidemia was present in the majority of patients before surgery and resolved or improved following surgery in almost all instances. CONCLUSIONS: The findings indicate that reliable and substantial weight loss can be accomplished by gastric bypass surgery with accompanying major reductions in associated co-morbidities. Such benefits suggest that greater attention should be given to this form of treatment for those with severe obesity. PMID- 11054248 TI - Energy and nitrogen absorption after biliopancreatic diversion. AB - BACKGROUND: The strict long-term weight maintenance in good nutritional conditions observed after biliopancreatic diversion (BPD) needs to be explained. MATERIALS AND METHODS: 15 operated subjects were maintained at an isoenergetic and isonitrogenic diet as similar as possible to their usual diet. Apparent absorption (AA) of energy, fat, nitrogen and calcium was calculated subtracting the fecal content, measured directly, from the oral intake, derived from tables. The alimentary protein absorption was directly determined by I125 albumin oral administration. RESULTS: Mean AA for energy and fat was 57% and 32%, respectively; AAs were unrelated as absolute value and negatively associated as percent of the intake with the energy and fat intake. I125 intestinal absorption was 73%, while nitrogen percent AA was 57%, indicating higher than normal loss of endogenous nitrogen. Calcium AA was 551 mEq/day, 26% of the intake. A positive correlation between nitrogen and calcium AA as absolute values and alimentary intake was observed, while there was no correlation when AA were considered as per cent of the intake. CONCLUSIONS: For energy and fat, an increase in intake corresponds to an increase in percent malabsorption, so that the absolute amount absorbed tends to remain constant, accounting for the excellent weight maintenance observed following BPD. This was confirmed by a long-term hypernutrition study after BPD. On the contrary, for nitrogen and calcium, the percent absorption tends to remain constant when intake varies, so that an increase in alimentary intake results in an increased absolute amount absorbed. PMID- 11054249 TI - Short-term changes in serum leptin concentration following biliopancreatic diversion. AB - BACKGROUND: The influence of the new anatomico-functional structure created by biliopancreatic diversion (BPD) in the postoperative fall of serum leptin concentration was evaluated. METHODS: Serum leptin concentration was determined in obese women before and immediately after BPD, before the usual postoperative intestinal rest. The measurements were repeated at the second postoperative month, when oral intake had nearly totally resumed and the patients had lost the first amount of weight. RESULTS: 5 days following BPD, a sharp reduction of serum leptin concentration was observed. At the second postoperative month the values remained nearly unchanged and were indistinguishable from those observed in a group of obese non-operated patients with a closely similar body weight. CONCLUSIONS: Changes in the upper gastrointestinal tract due to BPD appear to have no influence in the postoperative reduction of serum leptin concentration, which appears to be substantially related only to the patientis adiposity. PMID- 11054250 TI - BAROS: an effective system to evaluate the results of patients after bariatric surgery. AB - BACKGROUND: Success in bariatric surgery is most often evaluated by a sufficient loss of excess weight and an improvement in the medical conditions. The expected increase in quality of life (QoL) after weight loss, however, has not often been systematically analyzed. BAROS (Bariatric Analysis and Reporting Outcome System) is a scoring system which, along with easy handling, allows comparisons to be made internationally. METHODS: 386 morbidly obese patients who had undergone bariatric surgery in our hospital were evaluated with BAROS. Five categories- failure, fair, good, very good, excellent- were taken from the scoring system that BAROS offers. This system has three major points: excess weight loss, medical co-morbidities and QoL. Points are subtracted for reoperations and defined complications. The operations performed were silastic ring vertical gastroplasty (72%), adjustable gastric banding (23%), biliopancreatic diversion (3%), vertical banded gastroplasty (1%) and gastric banding of Molina (1%). RESULTS: In 1991 and 1993, we had a fair result in 3% and 10% of the patients. A good score with a mean of 3.6 to 4.1 was reached in 1992, 1994 and 1995. In the last 3 years, 1996 to 1998, the mean score was 5.0 to 5.7, which is a very good result when compared with the scoring key. CONCLUSION: BAROS is a valuable tool to access the QoL of patients who have undergone bariatric surgery. PMID- 11054251 TI - Dermolipectomies following weight loss after surgery for morbid obesity. AB - BACKGROUND: Dermolipectomies play a major role in the functional and esthetic deformities which result from massive weight loss. METHODS: From June 1994 to June 2000, 148 morbidly obese patients underwent various bariatric surgical procedures. After at least 1 year, 33 patients underwent 51 regional dermolipectomies performed by the same plastic surgeon. RESULTS: All 33 patients underwent abdominal dermolipectomy. The average operative time was 194.2 min (110 420 min). The average amount of tissue excised was 2948.6 g (850-7525 g). 4 patients (12.1%) required blood transfusion. 6 patients (18.1%) developed complications, which included 1 case of postoperative bleeding, 3 wound infections and 2 skin dehiscences. Average length of hospital stay was 9.5 days (5-22 days). 15 of these patients (45. 4%) simultaneously underwent abdominal incisional hernia repair; in 9 (24.2%), a Gore-Textrade mark mesh was used. In 2 patients the procedure was performed under emergency conditions due to small bowel obstruction. In 2 patients, simultaneous cholecystectomy was also performed. In 1 patient, a suction-assisted lipectomy of both thighs was necessary. 7 patients (21.2%) had mammaplasty, with average operative time 175.7 min (140-210 min). In 1 of them, breast implants were placed. There was no morbidity, and the average hospitalization was 6 days (4-9 days). Flankplasty was done in 4 patients (12.1%), thigh reduction plasty in 4 patients (12.1%), and arm reduction plasty in 3 patients (9%). The average operative time was 302.5 min (160-420), 246.2 min (230-280) and 203.3 min (180-240) respectively. Average tissue excised was 1503 g (725-2400 g), 1342.5 g (1050-1550 g), and 572.6 g (400 848 g), respectively. Morbidity was related to wound infection in 1 patient, and persistent edema of the left lower extremity in another. 4 of these 18 patients required blood transfusion. Average hospitalization was 8.2 days (6-11), 8 days (7-9) and 6 days (5-7) respectively. CONCLUSIONS: Regional dermolipectomies constitute the only available treatment for deformities following massive weight loss after bariatric surgery. Based on our experience, these procedures are safe, without serious complications and with good functional and esthetic results. PMID- 11054252 TI - Breast reconstruction facilitated by vertical banded gastroplasty. AB - BACKGROUND: Vertical banded gastroplasty (VBG) causes weight loss, which is often associated with redundant abdominal tissue. This redundant tissue can be used successfully for breast reconstruction or breast augmentation in case of mastectomy or ptotic hypotrophic breasts. METHOD: One patient with bilateral mastectomy is described in whom the weight fell from BMI 52 to BMI 26 after VBG, giving the patient an abdominal "apron", which facilitated bilateral breast reconstruction. RESULTS: Bilateral deep inferior epigastric perforator (DIEP) flap breast reconstruction after weight reduction following VBG resulted in an esthetic pleasing result with additional correction of the cutis laxa abdominis. CONCLUSION: Autologous breast reconstruction can be performed safely with the methods used today, after successful weight loss following obesity surgery. PMID- 11054253 TI - Organ transplantation and obesity: evaluation, risks and benefits of therapeutic strategies. AB - Obesity is a prevalent health problem that has discernible impact on all fields of surgery. However, little attention is paid in the literature to the underlying relation of surgical, immunological and metabolic links between transplantation and morbid obesity. Pre-operative obesity has been reported to worsen the outcome of organ transplantation. Impairment of graft function as well as decreased patient and graft survival can contribute to this effect. Post-transplant weight gain is common and may be attributed to an imbalance of the adipostatic and appetite stimulating hormones. Reduction of obesity before transplantation has to cope with limited time, increased risk of therapeutic side-effects in patients with end-stage organ failure, and psychosocial stress. Overweight reduction following organ transplantation interferes with diverse effects associated with immunosuppressive therapy. A case of adjustable gastric banding following renal transplantation is presented. PMID- 11054254 TI - Bowel obstruction after biliopancreatic diversion: a deceptive complication. AB - BACKGROUND: Although obesity surgery is now practiced in most of the world, many general surgeons, faced with an emergency, are not experienced in the diagnostic problems associated with these techniques, or about the most suitable treatment to resolve the acute pathology while preserving the weight loss. The biliopancreatic diversion (BPD), because of its complexity, could cause a delay in the diagnosis and therapy, with possible catastrophic consequences for the patient. METHODS: We report 3 patients with bowel obstruction after BPD. In the first patient intestinal occlusion was due to an adhesion obstructing the alimentary tract; in the other two patients the occlusion was localized to the biliopancreatic tract, due to a serrate stenosis of the entero-entero anastomosis in one patient and due to volvulus of the biliopancreatic loop in the other patient. RESULTS: Signs and symptoms were different according to whether the obstruction was in the alimentary tract or the biliopancreatic tract. In all cases a prompt gastrointestinal x-ray with barium and ultrasound scan and/or CT scan induced us to a mandatory laparotomy with resolution of the obstruction. CONCLUSIONS: After BPD, diagnosis of an intestinal obstruction must be made promptly. Even colleagues who express doubts must be persuaded to perform immediately an upper gastrointestinal tract x-ray and an U/S or CT scan. In this way, it may be possible to avoid intestinal resection and catastrophic complications. PMID- 11054255 TI - Intragastric erosion of laparoscopic adjustable silicone gastric band. AB - Intragastric erosion of the adjustable silicone gastric band (ASGB) is a rare but severe complication of gastric banding, often leading to reoperation. We describe our experience with 4 cases referred to us. The best timing of removal and the choice of another bariatric procedure is still controversial. We advise to wait until migration of the band into the lumen is complete. With removal of the ASGB if another weight reduction procedure is advisable, conversion to a biliopancreatic diversion is possible. PMID- 11054256 TI - Understanding the bariatric surgical patient: a demographic, lifestyle and psychological profile. AB - BACKGROUND: Although improved recently, the public is generally unaware of current scientific knowledge in bariatrics and the availability of surgery for massive obesity. As bariatric professionals we recognize the need for this education. Understanding the patientis socio-economic, demographic and psychological make-up and lifestyle preferences is a crucial element. METHODS: 1200 obesity surgery patients were sent questionnaires to assess a variety of personal parameters. 395 (33%) were returned and tabulated. A literature review of the obese person's psychological profile was also summarized. RESULTS: An overview of these patients' height, weight, age, ethnicity, marital, educational and employment status, number of children, residential population, and income are presented. Their television and movie viewing, radio listening, social event participation, print media and Internet habits are described. The psychological profile is outlined so that the whole person is understood. CONCLUSION: Understanding the obese person is primary to effectively educating the public. There are many appropriate and much-needed applications of this information to the public, the obese population, other health-care providers, legislators, and insurers. PMID- 11054257 TI - Making the most of your time: the benefits of converting patient education programs into continuing nursing education. AB - Educating hospital staff and patients is a time-consuming, yet essential component to a successful bariatric surgery program. The process of converting patient education and support group sessions into continuing education opportunities for hospital staff is described. This also provides hospital staff with direct exposure to patient testimony of concerns and obstacles to following treatment plans. PMID- 11054258 TI - Live attenuated HIV vaccines: a proposal for further research and development. PMID- 11054259 TI - Apparent founder effect during the early years of the San Francisco HIV type 1 epidemic (1978-1979). AB - HIV-1 envelope sequence variants were RT-PCR amplified from serum samples cryopreserved in San Francisco in 1978-1979. The HIV-1 subtype B env V3-V5 sequences from four homosexual men clustered phylogenetically, with a median nucleotide distance of 2.8%, reflecting a recent common origin. These early U.S. HIV-1 env variants mapped close to the phylogenetic root of the subtype B tree while env variants collected in the United States throughout the 1980s and 1990s showed, on average, increasing genetic diversity and divergence from the subtype B consensus sequence. These results indicate that the majority of HIV-1 currently circulating in the United States may be descended from an initial introduction and rapid spread during the mid- to late 1970s of subtype B viruses with limited variability (i.e., a founder effect). As expected from the starburst-shaped phylogeny of HIV-1 subtype B, contemporary U.S. strains were, on average, more closely related at the nucleic acid and amino acid levels to the earlier 1978 1979 env variants than to each other. The growing levels of HIV-1 genetic diversity, one of multiple obstacles in designing a protective vaccine, may therefore be mitigated by using epidemic founding variants as antigenic strains for protection against contemporary strains. PMID- 11054260 TI - Recovery of hematopoietic activity in bone marrow from human immunodeficiency virus type 1-infected patients during highly active antiretroviral therapy. AB - The mechanisms responsible for the hematopoietic failure in human immunodeficiency virus type 1 (HIV-1)-infected patients are still unknown. Several findings indicate that the in vitro proliferative potential of precursor cells from AIDS patients is reduced. The changes seen in bone marrow (BM) morphology and the defective BM functions associated with cytopenias have both been proposed as potential explanations. In patients treated with highly active antiretroviral therapy (HAART) an immune reconstitution associated with increased whole blood cell counts has been described. We have investigated the effects of HAART on the number of colony-forming cells (CFCs) and long-term culture initiating cells (LTC-ICs), using long-term BM cell cultures (LTBMC) in a group of subjects with HIV-1 infection enrolled in an open study to evaluate the mechanisms of immune reconstitution during HAART. In each patient, the increase in colony growth was homogeneous, regardless of the type of hematopoietic progenitor cells assayed; in four subjects an increase in the most primitive progenitor cells (LTC-ICs) was observed. These findings were associated with the in vivo data showing increased numbers of BM mononuclear cells (BMMCs) after HAART and with a rise in peripheral CD4(+) T cell counts and decreased levels of plasma HIV-1 RNA. A decreased number of hematopoietic progenitor cells and/or a defective modulation of progenitor cell growth might be the cause of the hematological abnormalities in AIDS patients. Controlling HIV-1 replication by HAART could determine a restoration of stem cell activity, probably because of the suppression of factors that inhibit normal hematopoiesis. PMID- 11054261 TI - Limited heterogeneity of HIV type 1 in infected mothers correlates with lack of vertical transmission. AB - The human immunodeficiency virus type 1 (HIV-1) envelope V3 region sequences of peripheral blood mononuclear cell DNA were analyzed from three nontransmitting mothers (infected mothers who failed to transmit HIV-1 to their infants in the absence of antiretroviral therapy), including one mother with two deliveries, and compared with the sequences of seven previously analyzed transmitting mothers. The coding potential of the envelope open reading frame, including several patient-specific amino acid motifs and previously described molecular features across the V3 region, were highly conserved. There was a low degree of heterogeneity within the sequences of each nontransmitting mother compared with the sequences of transmitting mothers. In addition, the estimates of genetic diversity of nontransmitting mother sequences were significantly lower compared with transmitting mother sequences. Phylogenetic analysis showed that the sequences of each nontransmitting mother formed distinct clusters that were well discriminated from each other and the sequences of seven transmitting mothers. In conclusion, a low degree of HIV-1 genetic heterogeneity in these infected mothers correlates with lack of vertical transmission; this finding may be useful in developing strategies for further prevention of maternal-fetal transmission. PMID- 11054262 TI - Evidence of a source of HIV type 1 within the central nervous system by ultraintensive sampling of cerebrospinal fluid and plasma. AB - Defining the source of HIV-1 RNA in cerebrospinal fluid (CSF) will facilitate studies of treatment efficacy in the brain. Four antiretroviral drug-naive adults underwent two 48-hr ultraintensive CSF sampling procedures, once at baseline and again beginning on day 4 after initiating three-drug therapy with stavudine, lamivudine, and nelfinavir. At baseline, constant CSF HIV-1 RNA concentrations were maintained by daily entry of at least 10(4) to 10(6) HIV-1 RNA copies into CSF. Change from baseline to day 5 ranged from -0.38 to -1.18 log(10) HIV-1 RNA copies/ml in CSF, and from -0.80 to -1.33 log(10) HIV-1 RNA copies/ml in plasma, with no correlation between CSF and plasma changes. There was no evidence of genotypic or phenotypic viral resistance in either CSF or plasma. With regard to pharmacokinetics, mean CSF-to-plasma area-under-the-curve (AUC) ratios were 38.9% for stavudine and 15.3% for lamivudine. Nelfinavir and its active M8 metabolite could not be accurately quantified in CSF, although plasma M8 peak level and AUC(0-8hr) correlated with CSF HIV-1 RNA decline. This study supports the utility of ultraintensive CSF sampling for studying HIV-1 pathogenesis and therapy in the CNS, and provides strong evidence that HIV-1 RNA in CSF arises, at least in part, from a source other than plasma. PMID- 11054263 TI - Development of a one-tube multiplex reverse transcriptase-polymerase chain reaction assay for the simultaneous amplification of HIV type 1 group M gag and env heteroduplex mobility assay fragments. AB - The emergence of intersubtype recombinant HIV-1 isolates has made it imperative to analyze different regions of HIV-1 genomes. For this purpose a one-tube multiplex RT-PCR, coamplifying first-round amplicons that allow amplification of gag and env heteroduplex mobility assay (HMA) fragments from different HIV-1 group M isolates, was developed, starting with plasma samples. The multiplex RT PCR assay is sensitive: 115 of 136 (84.5%) samples were positive for both gag and env, positive amplification of the gag fragment was observed in 130 of 136 (95.6%) samples, while for the env fragment 119 of 136 (87.5%) tested positive. The multiplex RT-PCR in combination with gag and env HMA makes large-scale HIV-1 subtyping fast, simple, and more economical. PMID- 11054264 TI - Evaluation of a second-generation nucleic acid sequence-based amplification assay for quantification of HIV type 1 RNA and the use of ultrasensitive protocol adaptations. AB - Accurate assessment of plasma HIV RNA levels at low concentrations is clinically important. We evaluated a second-generation quantitative HIV RNA assay (NucliSens HIV-1 QT), and three simple adaptations of the NucliSens standard protocol to lower the lower cutoff level. The assays were evaluated in constructed panels with known HIV RNA concentrations and in clinical samples. Results were compared with those obtained with the first generation (NASBA HIV-1 QT) and with two other commercially available assays: the Amplicor HIV Monitor test and the Quantiplex assay. In a constructed panel, results obtained by NASBA QT were on average 0.13 log(10) copies/ml (SD 0.15) higher than those of NucliSens. The NucliSens assay could quantify HIV RNA in at least 50% of the samples down to 518 (2.71 log(10)) copies/ml and NASBA QT to 5.80 x 10(3) (3.76 log(10)) copies/ml). Both assays correlated well with the known input (R NucliSens = 0.99; R NASBA QT = 0.996), but results were more variable at lower input levels. With the three different ultrasensitive NucliSens adaptations, HIV RNA could be quantified in at least 50% of the samples down to 100 (2.00 log(10)), 46 (1.66 log(10)), and 10 (1.00 log(10)) copies/ml, respectively. In patient samples, Amplicor results were on average 0.11 (SD 0.20) log(10) copies/ml above, NucliSens 0.02 (SD 0.29) copies/ml above, and Quantiplex 0.13 (SD 0.19) copies/ml below the mean of the three assay results per sample. The variation remained the same over the range of RNA levels with all three assays. The NucliSens assay can quantify HIV RNA at lower levels than the NASBA QT and is comparable to other commercially available assays. The lower cutoff of the NucliSens can be lowered down to 10 copies/ml. PMID- 11054265 TI - Calmodulin and HIV type 1: interactions with Gag and Gag products. AB - The level of calmodulin increases in cells expressing HIV-1 envelope glycoprotein. Although a calmodulin increase is bound to alter many cellular metabolic and signaling pathways, the benefits to the virus of these alterations must be indirect. However, the possibility exists that increased cellular calmodulin benefits the virus by directly associating with nonenvelope viral proteins. We have, therefore, investigated whether calmodulin can interact with HIV structural proteins Gag, p17, and p24. Calmodulin binds Gag and p17 but not p24 in (125)I-labeled calmodulin overlays of SDS-polyacrylamide gels. Removal of calcium by addition of EGTA eliminates this binding. A computer algorithm for predicting helical regions that should bind calmodulin predicts that there are two calmodulin-binding regions near the N terminus of p17. Intrinsic tryptophan fluorimetry shows that two peptides, each of which includes one of the predicted regions, bind calmodulin: p17(11-25) binds calmodulin with a 2-to-1 stoichiometry and dissociation constant of approximately 10(-9) M(2), and p17(31-46) also binds calmodulin with a dissociation constant of about 10(-9) M. These binding sites are nearly contiguous, forming an extended calmodulin-binding domain p17(11-46). In H-9 cells, Gag and calmodulin colocalize within the resolution of confocal light microscopy. PMID- 11054266 TI - Efavirenz is a potent nonnucleoside reverse transcriptase inhibitor of HIV type 1 replication in microglia in vitro. AB - The objective of this study was to determine whether reverse transcriptase inhibitors (RTIs) could decrease viral replication in microglia. Human microglia obtained from individuals undergoing temporal lobectomy were cultured and infected with HIV-1 isolates from the central nervous system (CNS) as previously described (Strizki JM, et al. J Virol 1996;70:7654-7662). These microglial cultures were treated with one of three nucleoside RTIs (NRTIs) or with efavirenz, a nonnucleoside RTI (NNRTI), at various time points before and during HIV-1 infection. The drug levels sufficient to provide > 90% inhibition of microglial HIV replication (IC90) were determined by comparison of p24(gag) release in the cultures among treated and untreated microglia. Infectious virus released from the infected cultures was also measured with U373-MAGI-CCR5 cells. Efavirenz, an NNRTI, blocked HIV-1(DS-br) infection of microglia with an IC(90) of 0.7-7 nM. This value is similar to the efavirenz IC(90) values for inhibition of laboratory and clinical isolates in lymphocytes, is 2-3 logs lower than the IC90 values of AZT and d4T, and is 1-2 logs lower than that of ddC in microglia. Efavirenz also inhibited infection with other neurotropic isolates, and with viruses isolated from other compartments that also replicated well in microglia. Thus, efavirenz is a potent inhibitor of HIV-1 infection in microglia. Furthermore, efavirenz IC(90) drug levels are present in the cerebrospinal fluid (CSF) of patients taking this once daily NNRTI. PMID- 11054267 TI - Overexpression of lymphocytic GD3 ganglioside and presence of anti-GD3 antibodies in patients with HIV infection. AB - This study was undertaken to analyze the role of disialoganglioside GD3 in HIV infection and disease progression. We report here the results obtained by both ex vivo and in vitro experiments on (1) surface and cytoplasmic expression and distribution of GD3 in HIV-infected cells, (2) the presence of anti-GD3 antibodies in sera of patients with HIV infection in various stages of the disease, and (3) the association of GD3 expression with HIV-related apoptotic events. GD3 expression was determined by high-performance thin-layer chromatography (HPTLC) and lipid-bound sialic acid and by static and flow cytometric analyses in peripheral blood lymphocytes from 22 AIDS patients, 20 anti-HIV Ab(+) asymptomatic subjects, and 25 healthy donors. Results obtained clearly indicated a significantly higher expression of plasma membrane GD3 content in lymphocytes from HIV-infected patients with respect to healthy controls. These HIV-induced perturbations of glycosphingolipid metabolism could be detected in all stages of the disease, including asymptomatic individuals. In addition, a significant percentage of patients showing disease progression displayed in serum samples an increased presence of anti-GD3 antibodies. Interestingly, ex vivo studies of lymphocytes from patients with HIV infection also indicated that GD3 expression is strictly associated with annexin V binding, an early marker of apoptosis. Moreover, cytofluorimetric analysis showed that virtually all anti-p24 Ab-positive cells were also immunolabeled with anti-GD3 antibodies. Accordingly, in vitro studies showed a significant redistribution and increase in GD3 expression in cultured U937 cells chronically infected with HIV-1 with respect to uninfected counterparts. In conclusion, our data clearly indicate that a significant increase in GD3 content in HIV-infected lymphocytes can occur and that this GD3 overexpression is paralleled by the presence of anti-GD3 antibodies in the plasma of patients. This is the first demonstration that disialoganglioside GD3, independent of the therapeutic schedule employed, can be considered as one of the early markers of HIV infection and can contribute to the early events leading to T cell depletion by apoptosis. PMID- 11054268 TI - A therapeutic HIV vaccine using coxsackie-HIV recombinants: a possible new strategy. AB - The ultimate goal in the treatment of HIV-infected persons is to prevent disease progression. A strategy to accomplish this goal is to use chemotherapy to reduce viral load followed by immunotherapy to stimulate HIV-specific immune responses that are observed in long-term asymptomatic individuals. An effective, live, recombinant virus, expressing HIV sequences, would be capable of inducing both CTL and CD4(+) helper T cell responses. To accomplish these goals, the viral vector must be immunogenic yet retain its avirulent phenotype in a T cell deficient host. We have identified a coxsackievirus variant, CB4-P, that can induce protective immunity against a virulent variant. In addition, the CB4-P variant remains avirulent in mice lacking CD4(+) helper T cells, suggesting that CB4-P may be uniquely suited as a viral vector for a therapeutic HIV vaccine. Two strategies designed to elicit CTL and CD4(+) helper T cell responses were used to construct CB4-P/HIV recombinants. Recombinant viruses were viable, genetically stable, and retained the avirulent phenotype of the parental virus. In designing a viral vector for vaccine development, an issue that must be addressed is whether preexisting immunity to the vector would affect subsequent administration of the recombinant virus. Using a test recombinant, we showed that prior exposure to the parental CB4-P virus did not affect the ability of the recombinant to induce a CD4(+) T cell response against the foreign sequence. The results suggest that a "cocktail" of coxsackie/HIV recombinants may be useful as a therapeutic HIV vaccine. PMID- 11054269 TI - CD8+ thymic lymphocytes express reduced levels of CD8beta and increased interferon gamma in cats perinatally infected with the JSY3 molecular clone of feline immunodeficiency virus. AB - Biological isolates of feline immunodeficiency virus (FIV) cause a relative expansion of activated single-positive CD8(+) (SP CD8(+)) lymphocytes within the thymus of infected cats. In this study, thymic SP CD8(+) lymphocytes were analyzed from cats inoculated as neonates with a pathogenic molecular clone of FIV, JSY3, which was previously derived from the wild-type biological isolate FIV(NCSU-1) (NCSU-1). Four cats were inoculated intraperitoneally with NCSU-1 and compared with 11 cats inoculated with JSY3. Five control cats matched in litter and age were administered an intraperitoneal sham inoculum. Between 12 and 16 weeks postinoculation, interferon-gamma (IFN-gamma) mRNA was quantified by RT-PCR in freshly isolated thymocytes and peripheral blood mononuclear cells (PBMCs). The quantity of IFN-gamma mRNA was increased more than 10-fold in thymocytes and PBMCs of 13 of 13 FIV-inoculated cats as compared with the sham-inoculated controls. IFN-gamma mRNA coenriched with magnetically sorted CD8(+) PBMCs and single-positive (SP) CD8(+) thymocytes. Cells expressing IFN-gamma mRNA were located within the thymic perivascular zone, along the corticomedullary junction, and adjacent to lymphoid follicles. The expansion of thymic SP CD8(+) cells was associated with an increase in CD8alpha(+)/beta(neg) and CD8alpha(+)/beta(lo) phenotypes, the latter population resembling a previously reported memory/effector peripheral blood cell with FIV suppressor activity. From these data we conclude that JSY3 and NCSU-1 induce similar phenotypic changes in thymic and peripheral blood CD8(+) cells. Thus, JSY3 is pathogenic for the thymus in vivo and will be useful for defining determinants of the CD8(+) cell response in this pediatric AIDS model. PMID- 11054270 TI - Characterization of immune escape viruses from a macaque immunized with live virus vaccine and challenged with pathogenic SHIVKU-1. AB - We characterized two immune escape viruses (SHIV(KU-1/105w52) and SHIV(KU 1/105w98)) from a macaque immunized with DeltavpuDeltanef SHIV-4 and challenged with pathogenic SHIV(KU-1). This macaque developed neutralizing antibodies as well as virus-specific CTLs against the challenge virus. However, the two new viruses could not be neutralized by anti-SHIV(KU-1)-specific neutralizing antibodies and were poorly recognized by challenge virus-specific CTLs. Sequence analysis of the gene encoding gp120 revealed several mutations in the protein that might have contributed to the development of the immune-escape viruses. PMID- 11054271 TI - William E. Stewart II. PMID- 11054272 TI - Interleukin-1/Toll receptor family members: receptor structure and signal transduction pathways. AB - Interleukin-1 (IL-1) is a central mediator of the inflammatory response. It plays a role in both systemic and local immune responses to invading microbes. There are two receptors (IL-1RI and IL-1RII) that mediate the cellular responses. These receptors belong to a family of receptors based on homologous receptor structure within the intracellular signaling domain. Other family members include the Drosophila protein Toll, the recently discovered mammalian Toll-like receptors (TLR), and the IL-18 receptor. Engagement of these receptors by their diverse ligands results in activation of very similar signal transduction cascades through use of common signaling intermediates. These signal transduction cascades lead to the activation of cellular responses that are known to regulate the innate immune response. Therefore, elucidating the function and redundancy of this receptor family is essential to the understanding of the innate immune response. This review examines each member of this receptor family and emphasizes similarities and potential differences in both receptor structure and signal transduction pathways to further the understanding of this complex receptor family. PMID- 11054273 TI - Pharmacokinetics and pharmacodynamics of IFN-beta 1a in healthy volunteers. AB - The pharmacokinetics of recombinant human interferon-beta1a (IFN-beta1a) (Rebif, Ares-Serono, Geneva, Switzerland) were investigated in healthy volunteers following intravenous (i.v.) administration of increasing single doses of the drug (22 microg/6 million international units [MIU], 44 microg/12 MIU, and 66 microg/18 MIU); i.v., intramuscular (i.m.), and subcutaneous (s.c.) administration of a 66-microg dose; and repeated s.c. administration of four 66 microg doses at 48-h intervals. The disposition of IFN-beta1a followed triexponential decay after i.v. administration (half-lives 3 min, 42 min, and 22 h, respectively). After s.c. and i. m. administration, absorption was the rate limiting factor in the terminal phase. The median absolute bioavailabilities were 30% and 27%, respectively. The accumulation ratio after repeated s.c. injections was 2.4, and a terminal half-life of 66 h was observed. Intracellular 2-5A synthetase activity and serum neopterin and beta2-microglobulin concentrations increased after all IFN-beta1a injections and remained elevated following every other-day administration. The local tolerance was good, and the systemic tolerance was satisfactory. PMID- 11054274 TI - Identification by electron microscopy of the maturation steps in vaccinia virus morphogenesis inhibited by the interferon-induced enzymes, protein kinase (PKR), 2-5A synthetase, and nitric oxide synthase (iNOS). AB - Interferons (IFN) play a major role as a first-line host defense mechanism against viral infections. As treatment of animal cells with IFN induces a large number of genes, it has been difficult to assign the role of these genes in the antiviral action of IFN. Vaccinia virus (VV) is an ideally suited system to study IFN action because all steps in viral morphogenesis can be followed easily by electron microscopy (EM) of ultrathin sections from infected cells. To define the role of IFN-induced genes in viral morphogenesis, we have independently expressed from VV recombinants in primary chicken embryo fibroblast (CEF) cells each of the three IFN-induced genes encoding protein kinase (PKR), 2-5A synthetase, and inducible nitric oxide synthase (iNOS). By EM analysis, we have identified the steps in VV morphogenesis that are affected by each of the IFN-induced enzymes in comparison with untreated and IFN-treated cells. We found that in cells pretreated with IFN and infected with VV, immature virus (IV) is formed, but further stages of maturation are blocked. In cells infected with a VV recombinant expressing PKR (VV-PKR), there is severe inhibition on virus factories, and only few IV are formed. In cells infected with a VV recombinant expressing 2-5A synthetase (VV-2-5A), VV assembly is inhibited at or after IV formation. In cells infected with a VV recombinant expressing iNOS (VV-iNOS), all stages in VV morphogenesis are observed but with aberrant forms. In addition to the effects on viral assembly, in cells infected with either VV-PKR, VV-2-5AS, or VV-iNOS, there is nucleus condensation characteristic of apoptosis. Our findings have identified the steps in VV morphogenesis inhibited by PKR, 2-5A, and iNOS, provided a distinction between these effects, and highlighted a functional redundancy of the IFN system to block viral infection and to induce apoptosis. PMID- 11054275 TI - cDNA cloning of biologically active chicken interleukin-18. AB - By searching a chicken EST database, we identified a cDNA clone that appeared to contain the entire open reading frame (ORF) of chicken interleukin-18 (ChIL-18). The encoded protein consists of 198 amino acids and exhibits approximately 30% sequence identity to IL-18 of humans and various others mammals. Sequence comparisons reveals a putative caspase-1 cleavage site at aspartic acid 29 of the primary translation product, indicating that mature ChIL-18 might consist of 169 amino acids. Bacterially expressed ChIL-18 in which the N-terminal 29 amino acids of the putative precursor molecule were replaced by a histidine tag induced the synthesis of interferon-gamma (IFN-gamma) in cultured primary chicken spleen cells, indicating that the recombinant protein is biologically active. PMID- 11054276 TI - A polymorphism in the promoter region of the gene for interleukin-6 is associated with susceptibility to type 1 diabetes mellitus. AB - Type 1 diabetes mellitus is an autoimmune disease characterized by the destruction of the insulin-producing islet beta cells. It is likely that several genetic and environmental factors contribute to this process. There is increasing evidence showing that polymorphisms in cytokine genes may play an important role in modifying the immune response. Interleukin-6 (IL-6) is a cytokine that has been implicated in a number of immune-mediated diseases. Further, there is a polymorphism at position -174 (G(-174)C) of the promoter region of the IL-6 gene that may alter the expression of the gene. In this study, the G(-174)C polymorphism was investigated in 257 Caucasoid patients with type 1 diabetes, 53 two-parent-proband trios, and 120 normal, healthy controls. DNA was amplified using amplimers that flank the G(-174)C site, and the products were digested with the restriction endonuclease NlaIII to detect the G or the C allele. The homozygous G,G(-174) genotype was increased in the patients compared with the normal controls (50.6% vs. 33.3%, p < 0.002), with a decrease in the C,C genotype in the patients compared with the controls (12.5% vs. 24.2%, respectively, p < 0.004). In the 53 trios studied, the G allele was transmitted in 29 of 53 informative meioses. There was no association with age at onset of diabetes or the presence of diabetic complications. In conclusion, these results suggest that the IL-6 gene may contribute to the genetic susceptibility to type 1 diabetes. PMID- 11054277 TI - Induction of porcine cytokine mRNA expression after DNA immunization and pseudorabies virus infection. AB - Injection of plasmid DNA encoding pseudorabies virus (PRV) glycoprotein into pig muscle has been shown to result in protective immunity against lethal infection. Here, pigs were vaccinated by a single coinjection of three plasmids encoding PRV glycoproteins gB, gC, and gD, with plasmid expressing porcine granulocytemacrophage colony-stimulating factor (GM-CSF) or porcine interferon alpha (IFN-alpha). DNA immunization induced a primary T cell-mediated response characterized by low rates of IFN-gamma, interleukin-2 (IL-2), and IL4 mRNA in peripheral blood mononuclear cells (PBMC). Very low rates of PRV-specific IgG1 and the absence of IgG2 were obtained. Codelivery of plasmid expressing GM-CSF or IFN-alpha had no effect on cytokine mRNA expression or on B cell response. After a high virulent challenge, high levels of cytokine mRNA, mainly IFN-gamma, and high secondary antibody (Ab) response were induced in all DNA-vaccinated pigs. Codelivery of GMCSF gene significantly increased both Th immune response (i.e., IFN-gamma and IL-4 mRNA expression) and clinical protection but had no effect on secondary B immune response. Codelivery of IFN-alpha gene had no beneficial effect on secondary T and B cell immune responses. PMID- 11054278 TI - Interleukin-12 differentially regulates expression of IFN-gamma and interleukin-2 in human T lymphoblasts. AB - Interleukin-12 (IL-12) is known to upregulate expression of interferon-gamma (IFN gamma) by activated T cells. However, the effects of IL-12 on production of other Th1-type cytokines are less well defined. In this study, we examined the effects of IL-12 on expression of several cytokines, including IFN-gamma, IL-2, tumor necrosis factor-alpha (TNF-alpha), and IL-10, by primary human CD3(+) T cells. Although purified resting T cells were largely nonresponsive to IL-12 stimulation, anti-CD3-activated T cell blasts were strongly responsive, as demonstrated by the ability of IL-12 to induce Stat4 DNA-binding activity. Restimulation of T lymphoblasts on immobilized anti-CD3 monoclonal antibodies (mAb) induced rapid expression of TNF-alpha mRNA and more gradual increases in mRNA levels for IL-2, IFN-gamma, and IL-10. IL-12 markedly upregulated expression of IFN-gamma and IL-10 but downregulated expression of IL-2 in a dose-dependent and time-dependent manner. The levels of IL-2 produced by IL-12-treated T cells correlated inversely with the levels of IL-10. Moreover, neutralization of IL-10 activity with anti-IL-10 antibodies normalized IL-2 production by IL-12-treated T cells, confirming that the inhibition of IL-2 production by IL-12 was IL-10 mediated. Thus, IL-12 amplified expression of IFN-gamma and IL-10 and, via its ability to upregulate production of IL-10, inhibited expression of IL-2. These findings demonstrate that IL-12 differentially regulates expression of the Th1 type lymphokines, IFN-gamma and IL-2, in T lymphoblasts. PMID- 11054279 TI - Microspheres containing neutralizing antibodies to tumor necrosis factor-alpha and interleukin-1 beta protect rats from Staphylococcus aureus-induced peritonitis. AB - Previous studies using microencapsulated neutralizing antibodies (NA) to tumor necrosis factor (TNF) and interleukin-1 (IL-1) in combination with gentamicin have demonstrated improved survival in a peritonitis model of gram-negative septic shock. Microencapsulation has been shown to improve the effectiveness of NA by delivering them intracellularly, taking advantage of the natural phagocytic activity of the macrophage. It is the purpose of this study to see if microencapsulated NA to TNF and IL-1 in combination with vancomycin can improve survival compared with NA in solution in Staphlococcus aureus-induced septic shock. Groups of 10 rats received the following treatments: (1) S. aureus plus no treatment, (2) S. aureus plus blank microspheres, (3) S. aureus plus vancomycin, (4) S. aureus plus a microsphere form of NA and vancomycin, (5) S. aureus plus a solution form of NA and vancomycin, (6) S. aureus plus a microsphere form of NA, and (7) S. aureus plus a solution form of NA. Survival was monitored for 5 days, and plasma TNF and IL-1 levels were measured for 48 h after S. aureus administration. All (100%) animals that received the microsphere form of NA plus vancomycin, 20%-70% of the animals that received the microsphere form of NA alone, and 20% of the animals that received antibiotics alone survived for 5 days or more. None of the animals in the no treatment group or blank microsphere treatment group and only 10% of the animals in the solution form of NA plus or minus vancomycin group survived for more than 5 days. Plasma TNF and IL-1 levels were significantly increased after S. aureus treatment. Simultaneous and delayed treatment with the microsphere form of NA plus or minus vancomycin significantly reduced TNF and IL-1 levels, and the solution form of NA significantly reduced only TNF levels after immediate treatment. The survival rate was higher in animals with lower TNF levels and IL-1 levels. The results demonstrate that the microsphere form of cytokine NA is 100% effective in combination with vancomycin in protecting rats from S. aureus-induced peritonitis. The microsphere form was also more efficient in attenuating both TNF and IL-1 levels. PMID- 11054280 TI - Endotoxin and cytokine regulation of toll-like receptor (TLR) 2 and TLR4 gene expression in murine liver and hepatocytes. AB - Toll-like receptor (TLR) 2 and TLR4 are members of the interleukin-1 receptor (IL 1R) family and transduce similar signals as IL-1R in response to bacteria and bacterial components. In this study, we investigated the regulation of their gene expression in murine tissues, especially in the liver and hepatocytes. When mice were administered lipopolysaccharide (LPS), TLR2 mRNA was upregulated in the brain, heart, lung, liver, and kidney. In contrast, it was downregulated in the spleen. TLR4 mRNA was decreased in the brain. In the heart and lung, it increased, and it was not affected in the liver, kidney, and spleen. TLR mRNA was further analyzed in the liver and hepatocytes. Like LPS treatment, administration of IL-1, IL-6, or tumor necrosis factor (TNF) upregulated TLR2 mRNA. However, none of them affected the TLR4 mRNA level. In primary cultured hepatocytes, TLR2 mRNA was upregulated by LPS, IL-1, or TNF but not by IL-6 or dexamethasone. None of them affected TLR4 mRNA expression. Similar responses were observed in the murine hepatoma cell line Hepa 1-6. These results suggest that in infection with gram-negative bacteria, LPS and proinflammatory cytokines differentially regulate gene expression of TLR2 and TLR4 in murine hepatocytes, which may lead to pathologic and host defense reactions in the liver. PMID- 11054282 TI - Evolution of the family of pRN plasmids and their integrase-mediated insertion into the chromosome of the crenarchaeon Sulfolobus solfataricus. AB - Plasmid pHEN7 from Sulfolobus islandicus was sequenced (7.83 kb) and shown to belong to the archaeal pRN family, which includes plasmids pRN1, pRN2, pSSVx and pDL10 that share a large conserved sequence region. pHEN7 is most closely related to pRN1 in this conserved region. It also shares a large variant region containing several homologous genes with pDL10, which is absent from the other plasmids. The variant region is flanked by the sequence motif TTAGAATGGGGATTC and similar duplicated motifs occur in plasmids pRN1 and pRN2, separated by a few bases. It is inferred that recombination at these sites produces the main genetic variability in the plasmid family. The conserved region of the plasmid, and duplicated copies of the motif, are also present in the genome of Sulfolobus solfataricus P2. Moreover, they are bordered by a partitioned integrase gene (int) and by a 45 bp perfect direct repeat corresponding to the downstream half of a tRNA(Val) gene. The integrase and the direct repeat are highly similar in sequence to the integrase and the chromosomal integration site (att), respectively, of the SSV1 virus, which integrates into the chromosome of Sulfolobus shibatae. Recombination at the att repeats in S. solfataricus would produce a novel plasmid, pXQ1, which carries both an intact integrase gene and a single integration site (att). This strongly suggests that the same mechanism of site-specific integration at a tRNA gene is used for both viruses and plasmids in Sulfolobus. PMID- 11054283 TI - Three-dimensional view of the surface motif associated with the P-loop structure: cis and trans cases of convergent evolution. AB - Here we identify the determinants of the nucleotide-binding ability associated with the P-loop-containing proteins, inferring their functional importance from their structural convergence to a unique three- dimensional (3D) motif. (1) A new surface 3D pattern is identified for the P-loop nucleotide-binding region, which is more selective than the corresponding sequence pattern; (2) the signature displays one residue that we propose is the determinant for the guanine-binding ability (the residues aligned to ras D119; this residue is known to be important only in the G-proteins, we extend the prediction to all the other P-loop- containing proteins); and (3) two cases of convergent evolution at the molecular level are highlighted in the analysis of the active site: the positive charge aligned to ras K117 and the arginine residues aligned to the GAP arginine finger. The analysis of the residues conserved on protein surfaces allows one to identify new functional or evolutionary relationships among protein structures that would not be detectable by conventional sequence or structure comparison methods. PMID- 11054284 TI - Functions of the subunits in the FlhD(2)C(2) transcriptional master regulator of bacterial flagellum biogenesis and swarming. AB - In enterobacteria like Salmonella, biogenesis of cell surface flagella needed for motility is dependent upon the master operon flhDC at the apex of the flagellar gene hierarchy. The operon products FlhD and FlhC act together in a FlhD(2)C(2 )heterotetramer to induce flagellar gene transcription, while FlhD also represses cell septation. The flhDC operon is pivotal to differentiation into elongated hyperflagellated swarm cells that undergo multicellular migration, most strikingly in Proteus. We set out to establish the mechanism of action of the FlhD(2)C(2) multimer. In Proteus swarm cell extracts, all the FlhC was assembled into the FlhD(2)C(2 )transcription activator, but FlhD additionally formed approximately equimolar amounts of a FlhD(2) homodimer. Both FlhD and FlhC subunits homodimerised in vivo and in vitro, suggesting that self-interactions stabilise the FlhD(2)C(2 )complex. The FlhC and FlhD subunit proteins were separately expressed and purified, and the FlhD(2)C(2)heterotetramer was reconstituted in vitro. Purified FlhC bound specifically and cooperatively to the promoter region of the flhDC-regulated flhB flagellar gene in the absence of FlhD. Purified FlhD was unable to bind this target DNA, but binding by the FlhD(2)C(2)complex was approximately tenfold greater than the FlhC subunit alone, suggesting that FlhD potentiated the FlhC/DNA interaction. In support of this possibility, pre-incubation of FlhC with FlhD reduced the apparent dissociation constant, K(D), for the FlhC/DNA complex from 100 nM to 13 nM. Furthermore, in competition assays, FlhD substantially increased the specificity of DNA recognition by FlhC, and also stabilised the resultant labile protein/DNA complex, prolonging its half-life from around two minutes to more than 40 minutes. FlhD(2)C(2)is therefore an atypical prokaryotic transcription activator in which interaction of the FlhC subunit with DNA target sequences is enhanced by the coexpressed helper subunit FlhD. PMID- 11054285 TI - The mitochondrial import receptor Tom70: identification of a 25 kDa core domain with a specific binding site for preproteins. AB - The mitochondrial import receptor of 70 kDa, Tom70, preferentially recognizes precursors of membrane proteins with internal targeting signals. We report the identification of a stably folded 25 kDa core domain located in the middle portion of Tom70 that contains two of the seven tetratricopeptide repeat motifs of the receptor. The core domain binds non-cleavable and cleavable preproteins carrying internal targeting signals with a specificity indistinguishable from the full-length receptor. Competition studies indicate that both types of preproteins interact with overlapping binding sites of the core domain and that at least one additional interaction site is present in the full-length receptor. We suggest a model of Tom70 function in import of membrane proteins whereby a hydrophobic preprotein concomitantly interacts with several binding sites of the receptor. PMID- 11054286 TI - Insights into the molecular recognition of the 5'-GNN-3' family of DNA sequences by zinc finger domains. AB - In order to construct zinc finger domains that recognize all of the possible 64 DNA triplets, it is necessary to understand the mechanisms of protein/DNA interactions on the molecular level. Previously we reported 16 zinc finger domains which had been characterized in detail to bind specifically to the 5'-GNN 3' family of DNA sequences. Artificial transcription factors constructed from these domains can regulate the expression of endogenous genes. These domains were created by phage-display selection followed by site-directed mutagenesis. A total of 84 mutants of a three-domain zinc finger protein have been analyzed for their DNA-binding specificity. Here, we report the results of this systematic and extensive mutagenesis study. New insights into zinc finger/DNA interactions were obtained by combining specificity data with computer modeling and comparison with known structural data from NMR and crystallographic studies. This analysis suggests that unusual cross-strand and inter-helical contacts are made by some of these proteins, and the general orientation of the recognition helix to the DNA is flexible, even when constrained by flanking zinc finger domains. These findings disfavor the utility of existing simple recognition codes and suggest that highly specific domains cannot be obtained from phage display alone in most cases, but only in combination with rational design. The molecular basis of zinc finger/DNA interaction is complex and its understanding is dependent on the analysis of a large number of proteins. This understanding should enable us to refine rapidly the specificity of other zinc finger domains, as well as polydactyl proteins constructed with these domains to recognize extended DNA sequences. PMID- 11054287 TI - Alternative design of a tRNA core for aminoacylation. AB - The core of Escherichia coli tRNA(Cys) is important for aminoacylation of the tRNA by cysteine-tRNA synthetase. This core differs from the common tRNA core by having a G15:G48, rather than a G15:C48 base-pair. Substitution of G15:G48 with G15:C48 decreases the catalytic efficiency of aminoacylation by two orders of magnitude. This indicates that the design of the core is not compatible with G15:C48. However, the core of E. coli tRNA(Gln), which contains G15:C48, is functional for cysteine-tRNA synthetase. Here, guided by the core of E. coli tRNA(Gln), we sought to test and identify alternative functional design of the tRNA(Cys) core that contains G15:C48. Although analysis of the crystal structure of tRNA(Cys) and tRNA(Gln) implicated long-range tertiary base-pairs above and below G15:G48 as important for a functional core, we showed that this was not the case. The replacement of tertiary interactions involving 9, 21, and 59 in tRNA(Cys) with those in tRNA(Gln) did not construct a functional core that contained G15:C48. In contrast, substitution of nucleotides in the variable loop adjacent to 48 of the 15:48 base-pair created functional cores. Modeling studies of a functional core suggests that the re-constructed core arose from enhanced stacking interactions that compensated for the disruption caused by the G15:C48 base-pair. The repacked tRNA core displayed features that were distinct from those of the wild-type and provided evidence that stacking interactions are alternative means than long-range tertiary base-pairs to a functional core for aminoacylation. PMID- 11054288 TI - Arginine side-chain dynamics in the HIV-1 rev-RRE complex. AB - The binding of human immunodeficiency virus type 1 (HIV-1) Rev protein to its viral RNA target, stem-loop IIB (SLIIB) within the Rev Response element (RRE), mediates the export of singly-spliced and unspliced viral mRNA from the nucleus to the cytoplasm of infected cells; this Rev-mediated transport of viral RNA is absolutely required for the replication of infectious virus. To identify important features that influence the binding affinity and specificity of this Rev-RRE interaction, we have characterized the arginine side-chain dynamics of the Rev arginine-rich motif (ARM) while bound to a 34 nt RNA oligomer that corresponds to SLIIB. As the specificity of the Rev-RRE interaction varies with salt concentration, arginine side-chain dynamics were characterized at two different salt conditions. Following NMR measurements of (15)N spin relaxation parameters for the arginine (15)N(epsilon) nuclei, the dynamics of the corresponding N(epsilon)-H(epsilon) bond vectors were interpreted in terms of Lipari-Szabo model-free parameters using anisotropic expressions for the spectral density functions. Results from these analyses indicate that a number of arginine side-chains display a surprising degree of conformational freedom when bound to RNA, and that arginine residues having known importance for specific RRE recognition show striking differences in side-chain mobility. The (15)N relaxation measurements at different salt conditions suggest that the previously reported increase in Rev-RRE specificity at elevated salt concentrations is likely due to reduced affinity of non-specific Rev-RNA interactions. The observed dynamical behavior of the arginine side-chains at this protein-RNA interface likely plays an important role in the specificity and affinity of Rev-SLIIB complex formation. PMID- 11054289 TI - Structures of l-fuculose-1-phosphate aldolase mutants outlining motions during catalysis. AB - The crystal structures of l-fuculose-1-phosphate aldolase (FucA) with and without a ligated analogue of dihydroxyacetone phosphate (DHAP) and of a number of active center mutants have resulted in a model of the catalytic mechanism. This model has now been confirmed by structural analyses of further mutations at the zinc coordination sphere and at the phosphate site. In addition, these mutants have revealed new aspects of the catalysis: the hydroxyl group of Tyr113' (from a neighboring subunit), which sits just outside the zinc coordination sphere, steers DHAP towards a productive binding mode at the zinc ion; Glu73 contacts zinc in between the two ligand positions intended for the DHAP oxygen atoms and thus avoids blocking of these positions by a tetrahedrally coordinated hydroxy ion; the FucA polypeptide does not assume its minimum energy state but oscillates between two states of elevated energy as demonstrated by a mutant in a minimum energy state. The back and forth motion involves a mobile loop connecting the phosphate site with intersubunit motions and thus with the Brownian motion of the solvent. The phosphate group is bound strongly at a given distance to the zinc ion, which prevents the formation of too tight a DHAP:zinc complex. This observation explains our failure to find mutants that accept phosphate-free substitutes for DHAP. The FucA zinc coordination sphere is compared with that of carbonic anhydrase. PMID- 11054290 TI - The 1.9 A resolution structure of phospho-serine 46 HPr from Enterococcus faecalis. AB - The histidine-containing phosphocarrier protein HPr is a central component of the phosphoenolpyruvate:sugar phosphotransferase system (PTS), which transfers metabolic carbohydrates across the cell membrane in many bacterial species. In Gram-positive bacteria, phosphorylation of HPr at conserved serine 46 (P-Ser-HPr) plays several regulatory roles within the cell; the major regulatory effect of P Ser-HPr is its inability to act as a phosphocarrier substrate in the enzyme I reaction of the PTS. In order to investigate the structural nature of HPr regulation by phosphorylation at Ser46, the structure of the P-Ser-HPr from the Gram- positive bacterium Enterococcus faecalis has been determined. X-ray diffraction analysis of P-Ser-HPr crystals provided 10,043 unique reflections, with a 95.1 % completeness of data to 1.9 A resolution. The structure was solved using molecular replacement, with two P-Ser-HPr molecules present in the asymmetric unit. The final R-value and R(Free) are 0.178 and 0.239, respectively. The overall tertiary structure of P-Ser-HPr is that of other HPr structures. However the active site in both P-Ser-HPr molecules was found to be in the "open" conformation. Ala16 of both molecules were observed to be in a state of torsional strain, similar to that seen in the structure of the native HPr from E. faecalis. Regulatory phosphorylation at Ser46 does not induce large structural changes to the HPr molecule. The B-helix was observed to be slightly lengthened as a result of Ser46 phosphorylation. Also, the water mediated Met51-His15 interaction is maintained, again similar to that of the native E. faecalis HPr. The major structural, and thus regulatory, effect of phosphorylation at Ser46 is disruption of the hydrophobic interactions between EI and HPr, in particular the electrostatic repulsion between the phosphoryl group on Ser46 and Glu84 of EI and the prevention of a potential interaction of Met48 with a hydrophobic pocket of EI. PMID- 11054291 TI - Deuterium-proton exchange on the native wild-type transthyretin tetramer identifies the stable core of the individual subunits and indicates mobility at the subunit interface. AB - Transthyretin is a human protein capable of amyloid formation that is believed to cause several types of amyloid disease, depending on the sequence deposited. Previous studies have demonstrated that wild-type transthyretin (TTR), although quite stable, forms amyloid upon dissociation from its native tetrameric form into monomers with an altered conformation. Many naturally occurring single-site variants of TTR display decreased stability in vitro, manifested by the early onset familial amyloid diseases in vivo. Only subtle structural changes were observed in X-ray crystallographic structures of these disease associated variants. In this study, the stability of the wild-type TTR tetramer was investigated at the residue-resolution level by monitoring (2)H-H exchange via NMR spectroscopy. The measured protection factors for slowly-exchanging amide hydrogen atoms reveal a stable core consisting of strands A, B, E, F, and interestingly, the loop between strands A and B. In addition, the faster exchange of amide groups from residues at the subunit interfaces suggests unexpected mobility in these regions. This information is crucial for future comparisons between disease-associated and wild-type tetramers. Such studies can directly address the regions of TTR that become destabilized as a consequence of single amino acid substitutions, providing clues to aspects of TTR amyloidogenesis. PMID- 11054292 TI - Free energy calculations on dimer stability of the HIV protease using molecular dynamics and a continuum solvent model. AB - Dimerization of HIV-I protease (HIV PR) monomers is an essential prerequisite for viral proteolytic activity and the subsequent generation of infectious virus particles. Disrupting dimerization of the enzyme can inhibit its activity. We have calculated the relative binding free energies between different dimers of the HIV protease using molecular dynamics and a continuum model, which we call MM/PBSA. We examined the dominant negative inhibition of the HIV PR by a mutated form of the protease and found relative dimerization free energies of homo- and hetero-dimerization consistent with experimental data. We also developed a rapid screening method, which was called the virtual mutagenesis method to consider other mutations which might stabilize non-wild-type heterodimers. Using this approach, we considered the mutations near the dimer interface which might cause dominant negative inhibition of the HIV PR. The rapid method we developed can be used in studying any ligand-protein and protein-protein interaction, in order to identify mutations that can enhance the binding affinities of the complex. PMID- 11054293 TI - C-terminal regions of Hsp90 are important for trapping the nucleotide during the ATPase cycle. AB - Hsp90 is an abundant molecular chaperone that functions in an ATP-dependent manner in vivo. The ATP-binding site is located in the N-terminal domain of Hsp90. Here, we dissect the ATPase cycle of Hsp90 kinetically. We find that Hsp90 binds ATP with a two-step mechanism. The rate-limiting step of the ATPase cycle is the hydrolysis of ATP. Importantly, ATP becomes trapped and committed to hydrolyze during the cycle. In the isolated ATP-binding domain of Hsp90, however, the bound ATP was not committed and the turnover numbers were markedly reduced. Analysis of a series of truncation mutants of Hsp90 showed that C-terminal regions far apart in sequence from the ATP-binding domain are essential for trapping the bound ATP and for maximum hydrolysis rates. Our results suggest that ATP binding and hydrolysis drive conformational changes that involve the entire molecule and lead to repositioning of the N and C-terminal domains of Hsp90. PMID- 11054294 TI - Structure of a mutant EF-G reveals domain III and possibly the fusidic acid binding site. AB - The crystal structure of Thermus thermophilus elongation factor G (EF-G) carrying the point mutation His573Ala was determined at a resolution of 2.8 A. The mutant has a more closed structure than that previously reported for wild-type EF-G. This is obtained by a 10 degrees rigid rotation of domains III, IV and V with regard to domains I and II. This rotation results in a displacement of the tip of domain IV by approximately 9 A. The structure of domain III is now fully visible and reveals the double split beta-alpha-beta motif also observed for EF-G domain V and for several ribosomal proteins. A large number of fusidic acid resistant mutations found in domain III have now been possible to locate. Possible locations for the effector loop and a possible binding site for fusidic acid are discussed in relation to some of the fusidic acid resistant mutations. PMID- 11054295 TI - The effects of matrices of paired substitutions in mid-acceptor stem on Drosophila tRNA(His) structure and end-processing. AB - End-maturation reactions, in which the 5' end leader and 3' end trailer of precursor tRNA are removed by RNase P and 3'-tRNase, respectively, are early, essential steps in eukaryotic precursor tRNA processing. End-processing enzymes may be expected to contact the acceptor stem of tRNA due to its proximity to both cleavage sites. We constructed matrices of pair-wise substitutions in mid acceptor stem at nt 3/70 and 4/69 of Drosophila tRNA(His) and analyzed their ability to be processed by Drosophila RNase P and 3'-tRNase. In accord with our earlier study of D/T loop processing matrices, we find that tRNA end processing enzymes respond to sequence changes differently. More processing defects were observed with 3'-tRNase than with RNase P, and substitutions at 4/69 reduced processing more than those at 3/70. We evaluated tRNA folding using structure probing nucleases and investigated the contribution of K(M) and V(Max) to the processing efficiency of selected variants. In one substitution (C3A), mis folding correlates with processing defects. In another (C69A), a disruption of structure appears to be transmitted laterally to both ends of the acceptor stem. Poor processing of C69A by RNase P is due entirely to a reduction in V(Max), but for 3'-tRNase, it is due to an increase in K(M). PMID- 11054296 TI - Direct evidence for the cooperative unfolding of cytochrome c in lipid membranes from H-(2)H exchange kinetics. AB - The interaction of cytochrome c (cyt c) with anionic lipid membranes is known to disrupt the tightly packed native structure of the protein. This process leads to a lipid-inserted denatured state, which retains a native-like alpha-helical structure but lacks any specific tertiary interactions. The structural and dynamic properties of cyt c bound to vesicles containing an anionic phospholipid (DOPS) were investigated by amide H-(2)H exchange using two-dimensional NMR spectroscopy and electrospray ionisation mass spectrometry. The H-(2)H exchange kinetics of the core amide protons in cyt c, which in the native protein undergo exchange via an uncorrelated EX2 mechanism, exchange in the lipid vesicles via a highly concerted global transition that exposes these protected amide groups to solvent. The lack of pH dependence and the observation of distinct populations of deuterated and protonated species by mass spectrometry confirms that exchange occurs via an EX1 mechanism with a common rate of 1(+/-0.5) h(-1), which reflects the rate of transition from the lipid-inserted state, H(l), to an unprotected conformation, D(i), associated with the lipid interface. PMID- 11054297 TI - Homology among (betaalpha)(8) barrels: implications for the evolution of metabolic pathways. AB - We provide statistically reliable sequence evidence indicating that at least 12 of 23 SCOP (betaalpha)(8) (TIM) barrel superfamilies share a common origin. This includes all but one of the known and predicted TIM barrels found in central metabolism. The statistical evidence is complemented by an examination of the details of protein structure, with certain structural locations favouring catalytic residues even though the nature of their molecular function may change. The combined analysis of sequence, structure and function also enables us to propose a phylogeny of TIM barrels. Based on these data, we are able to examine differing theories of pathway and enzyme evolution, by mapping known TIM barrel folds to the pathways of central metabolism. The results favour widespread recruitment of enzymes between pathways, rather than a "backwards evolution" model, and support the idea that modern proteins may have arisen from common ancestors that bound key metabolites. PMID- 11054299 TI - Bilateral simultaneous LASIK. PMID- 11054301 TI - Correlation between eyes in bilateral LASIK. PMID- 11054303 TI - Ocular explosions from periocular anesthetic injections. PMID- 11054306 TI - Author's reply PMID- 11054304 TI - Nonvitrectomizing vitreous surgery. PMID- 11054307 TI - Ocular explosions from periocular anesthetic injections. PMID- 11054308 TI - Ocular explosions from periocular anesthetic injections. PMID- 11054309 TI - Application of acellular diurnal allografts. PMID- 11054311 TI - Central corneal pachymetry and LASIK. PMID- 11054314 TI - The challenge of teaching ophthalmology: a Residency Review Committee perspective. PMID- 11054315 TI - The doctor's eye: seeing through the myopathy of congenital ptosis. PMID- 11054316 TI - Superior rectus-levator synkinesis: a previously unrecognized cause of failure of ptosis surgery. AB - PURPOSE: To describe a previously unreported type of ptosis associated with abnormal synkinesis between the superior rectus muscle and the levator palpebrae superioris. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Seven cases with congenital or longstanding unilateral ptosis presenting to a regional, tertiary referral, oculoplastic service. Six of these cases were seen within a period of 2 years. METHODS: Detailed observations of eyelid, ocular, and pupil movements of both eyes were performed before the planning of ptosis surgery anterior levator resection. MAIN OUTCOME MEASURES: Magnitude of ptosis and its variation with the position of gaze. RESULTS: Ptosis present in the primary position disappeared or markedly reduced with upgaze so that measurements of levator function were apparently normal. Close examination of the relative movement of the eyelids revealed evidence of superior rectus to levator synkinesis occurring during upgaze. In three cases the synkinesis was recognized only after failed ptosis surgery. Once recognized, two of these cases underwent further surgery with an excellent result. Three other patients all had successful surgery. CONCLUSIONS: Superior rectus to levator synkinesis may be easily overlooked if eyelid elevation in upgaze is ascribed to normal levator function rather than a synkinetic movement. We draw attention to the importance of identifying this relatively common condition to plan appropriate ptosis surgery. The lack of levator muscle tone in the primary position of gaze means that an augmented resection of the levator muscle should be performed. PMID- 11054317 TI - Effective small-incision surgery for involutional lower eyelid entropion. AB - OBJECTIVE: The aim of this study was to develop an effective and minimally invasive operation to correct lower eyelid entropion that would address both the horizontal and vertical laxity. DESIGN: A prospective, noncomparative, interventional case series. PARTICIPANTS: Thirty-five consecutive patients with involutional entropion, aged 62 to 92 years (mean, 77.1 years), had surgery on 45 lower eyelids. Of the 45 procedures, 33 (73%) had a primary procedure and 12 (27%) were reoperations. INTERVENTION: A lateral tarsal strip with diagonal tightening of the orbital septum and lower lid retractors to the lateral orbital rim was performed via a 1-cm lateral canthal incision. MAIN OUTCOME MEASURES: Complications and surgical outcome were monitored clinically for between 12 and 24 months after surgery. RESULTS: The results were analyzed from 42 eyelids (33 patients) with a mean follow-up of 17.1 months (range 12-24 months). Two patients died and one dropped out of the study 3 months after the second eyelid operation. In 36 cases (86%), the entropion was cured. Transient lateral orbital rim tenderness was noted in six cases (14%), and one patient had a wound infection. Anatomic recurrences were detected in six eyelids of six patients, and five of these (83%) were asymptomatic. CONCLUSIONS: This surgical approach has been found effective in 86% of eyelids. Adequate clinical followup has proven essential for accurate evaluation of entropion surgery. PMID- 11054318 TI - Floppy eyelid syndrome and mental retardation. AB - OBJECTIVE: To report four subjects with a combination of floppy eyelid syndrome, mental retardation, and increased mechanical stimulus to the affected side. DESIGN: Retrospective, noncomparative case series. PARTICIPANTS: The authors retrospectively reviewed the charts of four mentally retarded subjects with floppy eyelid syndrome. INTERVENTION: Surgical tightening of three upper lids and one lower lid was performed in three subjects. MAIN OUTCOME MEASURE: Relief of symptoms. RESULTS: The affected eyelids were treated surgically; in case 1 by anterior lamellar repositioning and lateral and medial canthal tightening, in case 2 by horizontal upper lid shortening, and in case 3 by horizontal lid shortening of both upper and lower lids. There was marked relief from symptoms in all three cases. In case 4, surgery was deferred at parental request. CONCLUSIONS: These cases support the role of mechanical factors in the pathogenesis of floppy eyelid syndrome. Subjects with mental retardation may cooperate poorly with examination, and we believe that there should be a careful search for floppy eyelid syndrome in the presence of chronic conjunctivitis or unexplained epitheliopathy. PMID- 11054319 TI - Granular cell tumor of the lacrimal sac and nasolacrimal duct: no invasive behavior with incomplete resection. AB - OBJECTIVE: Granular cell tumors are rare tumors of the periorbital region, and we present the second such case in the lacrimal sac. The natural history and clinical behavior of these tumors is reviewed. DESIGN: Case report. METHODS: The clinical presentation, workup, surgical approach, and pathology of a case of granular cell tumor of the lacrimal sac are presented. RESULTS: There has been no recurrence of the lacrimal sac tumor with incomplete resection. CONCLUSIONS: This case history further adds to the understanding of the biologic behavior of granular cell tumors in the periorbital region. PMID- 11054320 TI - The ice test versus the rest test in myasthenia gravis. AB - OBJECTIVE: To compare the ice test with the rest test in subjects with myasthenic and nonmyasthenic ptosis. DESIGN: Randomized, noninterventional trial. PARTICIPANTS: (1) Ten subjects with ptosis from previously undiagnosed myasthenia gravis. (2) Fifteen subjects with nonmyasthenic ptosis. METHODS: Application of ice compared with rest. MAIN OUTCOME MEASURES: Improvement in eyelid elevation in millimeters after the application of a surgical glove filled with ice or cotton. RESULTS: In myasthenic subjects, the median improvement of ptosis with the rest test was 2 mm and with the ice test was 4.5 mm. The difference between the rest and ice tests is significant (P: < 0.001). There was no improvement in ptosis in nonmyasthenic subjects with either test. CONCLUSION: In myasthenic ptosis, improvement in eyelid elevation after the ice test is in part caused by rest. The ice test significantly improves ptosis more than rest alone does. PMID- 11054322 TI - Discussion by marilyn B. Mets, MD PMID- 11054321 TI - Strabismus examination by telemedicine. AB - OBJECTIVE: To evaluate the reliability of strabismus assessment using telemedicine (TM) technology. DESIGN: Two prospective interobserver agreement studies. One study compared the agreement between a standard and a TM examination, whereas the other assessed agreement between two independent standard examinations. PARTICIPANTS: Strabismus patients over 4 years of age examined at a remote community clinic and patients assessed in a strabismologist's urban practice. METHODS: Forty-two patients were examined in person by a pediatric ophthalmologist at the remote community and independently by a pediatric ophthalmology fellow by means of TM (TM-standard study). The TM examination was performed with the help of a qualified ophthalmic assistant at the remote telecommunication center using a Power Cam 100 camera, a Picture Tel Concorde 4500 teleconferencing system, and a 224 kilobyte bandwidth. For comparison, independent in person examinations were performed on 43 patients by both examiners (standard-standard study). Agreement was measured using unweighted kappa (k) for categorical data, the intraclass correlation coefficient (ICC) for continuous data, and percent agreement. The odds of disagreement with TM (comparing the TM-standard versus standard-standard studies) was assessed with logistic regression analysis. MAIN OUTCOME MEASURES: Three parameters were assessed: (1) category of strabismus, determined by observation without cover test; (2) angle of deviation at 0.33 and 6.0 m; and (3) ocular muscle action. RESULTS: Agreement on the category of strabismus was good (k > 0.61) other than for vertical deviations. However, there was good to excellent agreement between TM and standard examinations on the vertical (ICC = 0.78) and horizontal (ICC = 0.79) angles of deviation with 6-m fixation with the cover test. Muscle ratings agreed within one point for the lateral, superior, and inferior rectus muscle actions in more than 90% of the eyes examined. Although good agreement was observed in the TM-standard study, it was inferior to the agreement in the standard-standard study. Examination by TM increased the odds of disagreement compared with examination in person by twofold to threefold. CONCLUSIONS: Strabismus examination can be performed with a good level of reliability with the use of medium bandwidth video teleconferencing equipment. However, reduced reliability has been noted in the detection of small vertical deviations by inspection and in evaluating oblique muscle actions. PMID- 11054323 TI - Intraocular penetration of tamoxifen. AB - OBJECTIVE: To document ocular penetration of oral tamoxifen in patients being administered systemic therapy by measuring intraocular and serum levels of the drug in a series of patients undergoing elective ocular surgery. DESIGN: Nonrandomized, prospective, comparative trial. PARTICIPANTS: Twenty-one eyes of 21 patients undergoing elective ocular surgery (cataract extraction or vitrectomy). Twenty patients were using the antiestrogen, tamoxifen, and one participant was not. Nine patients were excluded in the final analysis because of inadequate sample size. INTERVENTION: Preoperative serum samples and perioperative aqueous samples, vitreous samples, or both were obtained for each patient, and these were analyzed for tamoxifen and its metabolites. Dilated fundus examination was performed before surgery on all patients. MAIN OUTCOME MEASURES: Evidence of tamoxifen, its metabolites, or both in the samples. RESULTS: Tamoxifen was detected in all analyzed serum samples (range, 82.4-290.0 ng/ml.) from patients taking the medication and was found to have penetrated into both vitreous (range, 0.5-7.8 ng/ml) and aqueous (range, 0.5-3.9 ng/ml) cavities. No relationship was found between serum and intraocular levels. CONCLUSIONS: This study shows that tamoxifen penetrates intraocular fluids to varying degrees. The drug levels in aqueous and vitreous do not appear to correlate with serum levels. Evidence of tamoxifen retinopathy or keratopathy was not seen. PMID- 11054324 TI - Indocyanine green-assisted peeling of the retinal internal limiting membrane. AB - OBJECTIVE: To determine whether indocyanine green (ICG) stains and facilitates peeling of the retinal internal limiting membrane (ILM). To investigate the different staining properties of the posterior cortical hyaloid, retinal ILM, and the retina after ILM removal. DESIGN: Autopsy eye study. MATERIALS: Eleven human cadaveric eyes. METHODS: Open sky vitrectomy including removal of the posterior cortical vitreous was performed. A 0.5% ICG solution was then injected into the posterior vitreous cavity over the macula. The dye was allowed to settle on the macula for 5 minutes and was then removed by mechanical aspiration. Peeling of the ILM was initiated with a bent needle and completed with intraocular forceps. Specimens were submitted for light and electron microscopy. MAIN OUTCOME MEASURES: Staining properties and ease of peeling of retinal ILM were evaluated. Retinal ILM removal was confirmed by histopathologic and electron microscopic examination. RESULTS: ICG contact with the retinal surface resulted in bright green staining of the ILM. This stain greatly facilitated ILM peeling by improving direct visualization of the membrane. The underlying retina did not stain, thus providing a clear distinction between the stained ILM and the unstained retina. Continuous circular peeling of the ILM was easily completed with this technique. Light microscopic and ultrastructural studies confirmed removal of the ILM. CONCLUSIONS: ICG solution distinctly stains the nearly invisible retinal ILM in human cadaveric eyes. ICG staining greatly facilitates ILM peeling by providing a stark contrast between the stained ILM and the unstained retina. PMID- 11054325 TI - Posterior segment manifestations of active ocular syphilis, their response to a neurosyphilis regimen of penicillin therapy, and the influence of human immunodeficiency virus status on response. AB - OBJECTIVE: To determine the relative frequencies of signs in posterior segment ocular syphilis, the response to a neurosyphilis regimen of penicillin, and differences in findings between human immunodeficiency virus (HIV)-coinfected and -noncoinfected patients in a community setting. DESIGN: Retrospective, noncomparative, consecutive case series. PARTICIPANTS: Fourteen consecutive patients with posterior segment ocular syphilis over a 14-year period within or during the acquired immune deficiency syndrome era. INTERVENTION: Neurosyphilis intravenous penicillin regimen. MAIN OUTCOME MEASURES: Initial and final visual acuity; treponemal and nontreponemal serologic analyses; cerebrospinal fluid cell count, protein, and Venereal Disease Research Laboratory analyses; posterior segment signs; and relapses and recurrences. RESULTS: Blacks and males were predominantly affected. Five (36%) of patients were HIV coinfected, and ocular syphilis led to the HIV infection diagnosis in three. Four (29%) patients had received previous antibiotic therapy for primary or secondary syphilis, raising the suspicion of relapse. Two patients had negative nontreponemal serologic results. All patients responded rapidly to neurosyphilis therapy. One patient subsequently relapsed after neurosyphilis therapy, and a second was reinfected with recurrence of ocular involvement. One previously undescribed retinal manifestation was discovered: a sectorial retinochoroiditis with delayed retinal circulation in the involved area. CONCLUSIONS: Ocular syphilis is a form of neurosyphilis and requires neurosyphilis therapy regardless of when it develops after primary infection. Conventional syphilis staging is of little use in understanding ocular syphilis. A high suspicion for this diagnosis is appropriate, especially in poorer black males with posterior segment inflammatory disease. Human immunodeficiency virus coinfection with ocular syphilis is common, but does not affect response to a neurosyphilis regimen of penicillin in the short term. Awareness of the multiple presentations of posterior segment ocular syphilis will aid ophthalmologists in averting misdiagnosis or delayed diagnosis. PMID- 11054326 TI - Fluocinolone acetonide sustained drug delivery device to treat severe uveitis. AB - PURPOSE: Uveitis is often a chronic disease requiring long-term medical therapy. In this report, we describe a pilot safety and efficacy trial of a novel sustained drug delivery system containing fluocinolone acetonide to treat patients with severe uveitis. DESIGN: Prospective, noncomparative, interventional case series PARTICIPANTS: Patients with severe uveitis. METHODS: Sustained drug delivery devices designed to release fluocinolone acetonide for at least 2.5 years were implanted through the pars plana into the vitreous cavity of seven eyes of five patients. All patients had severe uveitis not well controlled with, or intolerant to, repeated periocular corticosteroid injections, systemic corticosteroids, nonsteroidal immunosuppressive agents, or a combination thereof at the time of device implantation. Before device implantation, patients underwent complete evaluation including history, ophthalmologic examination, fluorescein angiography, visual field testing, and electroretinography. After surgery, patients were reexamined at 1 week, 2 weeks, 4 weeks, and at 1- to 3 month intervals. Visual fields, electroretinograms, and fluorescein angiography were repeated at 3- to 6-month intervals. MAIN OUTCOME MEASURES: Preoperative and postoperative visual acuity, ocular inflammation, anti-inflammatory medication use, and intraocular pressure. RESULTS: Patients had a diagnosis of Behcet's syndrome (two eyes), or idiopathic panuveitis (five eyes, including two with necrotizing retinitis, two with progressive chorioretinitis, and one with iridocyclitis and intermediate uveitis). Patients were observed an average of 10 months (range, 5-19 months). All eyes had stabilized or improved visual acuity after device implantation, and four of seven eyes had an improvement of three lines or more. The mean initial visual acuity, measured by Snellen chart, was 20/207, and the mean final visual acuity was 20/57 (P = 0.02). After surgery, at the final visit, no eye had clinically detectable inflammation, and all seven eyes had a marked reduction in systemic, topical, and periocular anti inflammatory medication use. Four eyes had increased intraocular pressure 6 weeks to 6 months after device implantation. Intraocular pressure has been controlled on topical medications. No patient experienced intraoperative complications. CONCLUSIONS: A fluocinolone acetonide sustained drug delivery device is a promising new therapy for the treatment of severe uveitis. Intraocular pressure must be carefully monitored long after device implantation. Based on these data, a randomized study of a larger group of patients is warranted. PMID- 11054327 TI - Ketorolac versus prednisolone versus combination therapy in the treatment of acute pseudophakic cystoid macular edema. AB - OBJECTIVE: To evaluate the efficacy of ketorolac tromethamine 0.5% ophthalmic solution, prednisolone acetate 1.0% ophthalmic solution, and ketorolac and prednisolone combination therapy in the treatment of acute, visually significant, cystoid macular edema (CME) occurring after cataract extraction surgery. DESIGN: Randomized, double-masked, prospective trial. PARTICIPANTS: Twenty-eight patients who had undergone cataract extraction and in whom clinical CME developed within 21 to 90 days after cataract surgery. METHODS: Patients were randomized to topical therapy with ketorolac (group K), prednisolone (group P), or ketorolac and prednisolone combination therapy (group C) four times daily. Treatment was continued until CME resolved or for 3 months, whichever occurred first. Treatment was then tapered over 3 weeks. Examinations were monthly and included Snellen visual acuity, contrast sensitivity, Amsler grid, slit-lamp examination, dilated fundus examination, and fluorescein angiography. RESULTS: Twenty-six of 28 patients completed the study. Patients were enrolled an average of 48 days after surgery. The average improvements in Snellen visual acuity were as follows: 1.6 lines in group K, 1.1 lines in group P, and 3.8 lines in group C. This reached statistical significance for all visits when group C was compared with group P, and for visits 4 and 5 when group C was compared with group K. Group C reached a mean change of two lines or more by visit 2; at no time did either group K or P reach a mean two-line improvement. At no time was a significant difference detected between group K and P with regard to visual acuity or change from baseline. A two-line or more improvement in Snellen acuity was achieved in 16 of 26 patients (61%). Analysis by group revealed four of eight patients (50%) in group P, six of nine patients (67%) in group K, and eight of nine patients (89%) in group C who had achieved a two-line or more improvement. In patients who did improve two lines or more, improvement occurred an average of 2.75 months after initiating therapy in group P, 1.43 months in group K, and 1.33 months in group C. Improvements in contrast sensitivity and leakage on fluorescein angiography tended to mirror improvements in Snellen acuity. CONCLUSIONS: Treatment of acute, visually significant pseudophakic CME with ketorolac and prednisolone combination therapy appears to offer benefits over monotherapy with either agent alone. Patients were more likely to experience recovery of two lines or more of visual acuity. Patients treated with combination therapy or ketorolac monotherapy responded more quickly than did patients treated with prednisolone alone. PMID- 11054328 TI - Discussion by allan J. Flach, MD PMID- 11054329 TI - Cataract extraction with multifocal intraocular lens implantation: a multinational clinical trial evaluating clinical, functional, and quality-of-life outcomes. AB - OBJECTIVE: Two million cataract extractions are performed annually in the United States. The procedure is nearly always accompanied by implantation of a monofocal intraocular lens (IOL), which corrects the patient's distance vision. The authors' objective was to measure visual function and quality-of-life outcomes associated with bilateral implantation of a multifocal IOL, which corrects distance and near vision, and to compare the outcomes with those of the standard therapy. DESIGN: A prospective, randomized, double-masked, clinical trial was conducted at eight sites in the United States, seven sites in Germany, and one site in Austria. PARTICIPANTS: Participants included 245 cataract patients, 127 of whom received the multifocal IOL bilaterally and 118 of whom received a monofocal IOL of nearly identical construction bilaterally. METHODS: Clinical data included visual acuity (VA), complications, and adverse events. Quality-of life data were collected using a previously validated survey instrument at baseline, after first eye surgery, and after second eye surgery. RESULTS: At 3 months after surgery, patients who had received multifocal IOLs had significantly better uncorrected and distance corrected binocular near VA compared with patients who had received monofocal IOLs (mean uncorrected VA, 20/26 multifocal vs. 20/40 monofocal; mean distance corrected VA, 20/28 multifocal vs. 20/45 monofocal; P < 0.0001). Additionally, 96% of patients who had received multifocal IOLs and 65% of patients who had received monofocal IOLs achieved both 20/40 and J3 (Jaeger) or better uncorrected, binocular distance and near visual acuities (P < 0. 0001). Patients who had received multifocal IOLs were more likely than patients who had received monofocal IOLs to never wear glasses overall (32% multifocal vs. 8% monofocal; P < 0.0001). On a 4-point scale, patients who had received multifocal IOLs on average reported having between "a little bit" and "some" glare or halo, whereas patients who had received monofocal IOLs reported between "none" and "a little bit" of glare or halo (1.57 vs. 0.43; P < 0.001). Patients who had received multifocal IOLs rated their vision without glasses better overall at near and at intermediate distances (P < or = 0.002) and demonstrated better visual function for near tasks and social activities. CONCLUSIONS: Cataract patients who received multifocal IOLs at time of surgery obtained better uncorrected and distance corrected near VA and reported better overall vision, less limitation in visual function, less spectacle dependency, and more glare or halo than those who received traditional monofocal IOLs. PMID- 11054330 TI - Regular and irregular astigmatism after superior versus temporal scleral incision cataract surgery. AB - OBJECTIVE: To evaluate the effect of superior and temporal scleral incisions on regular and irregular astigmatism in small incision cataract surgery. DESIGN: Prospective, randomized, comparative clinical trial. PARTICIPANTS: One hundred seventy-four eyes of 87 patients with bilateral cataracts scheduled to undergo routine cataract surgery. METHODS: One eye of each patient was randomly assigned to the superior incision group, and the contralateral eye was allocated to the temporal incision group. Phacoemulsification and intraocular lens implantation were performed through an unsutured 4.1-mm scleral incision. Patients were examined 1 day and 1, 3, and 6 months after surgery. MAIN OUTCOME MEASURES: Surgically-induced regular astigmatism calculated with vector analysis method, irregular astigmatism obtained by Fourier analysis of videokeratography data, and uncorrected and corrected visual acuity. RESULTS: Postoperatively, the superior incision group showed slight against-the-rule astigmatic changes, whereas slight with-the-rule astigmatism was seen in the temporal incision group. The amount of against-the-wound astigmatism and absolute value of length of the induced vector did not differ significantly between groups (P > 0.05, paired t test). In both groups, irregular astigmatism 1 day after surgery was significantly greater than the preoperative levels (P < 0.001), but not thereafter. No significant intergroup difference was observed in the amount of irregular astigmatism at any postoperative visits (P > 0.05). There was no significant difference in uncorrected and corrected visual acuity between groups postoperatively (P > 0.05, chi-square test). CONCLUSIONS: In small scleral incision cataract surgery, superior and temporal approaches are comparable in terms of visual rehabilitation and induction of regular and irregular astigmatism. PMID- 11054331 TI - Injectable versus topical anesthesia for cataract surgery: patient perceptions of pain and side effects. The Study of Medical Testing for Cataract Surgery study team. AB - OBJECTIVE: To compare patient reports of intraoperative pain and postoperative side effects by different anesthesia strategies for cataract surgery. DESIGN: Prospective cohort study. PARTICIPANTS: Men and women 50 years of age and older undergoing 19,250 cataract surgeries at nine centers in the United States and Canada from June 1995 through June 1997. INTERVENTION: Topical anesthesia or anesthesia with injection, with or without sedatives, opioid analgesia, hypnotics, and diphenhydramine (Benadryl). MAIN OUTCOME MEASURES: Patient ratings of intraoperative pain, satisfaction with pain management, and early postoperative side effects (drowsiness, nausea, vomiting, or a combination thereof). RESULTS: Twenty-six percent of surgeries were performed using topical anesthesia alone, and the remainder were performed with peribulbar, retrobulbar, or facial nerve block, or a combination thereof. Local anesthesia by injection with sedatives and diphenhydramine resulted in the lowest reporting of any intraoperative pain (1.3%), with postoperative drowsiness (9.6%) and nausea, vomiting, or both (1.5%) comparable with those administered topical anesthesia alone. Among those receiving topical anesthesia, use of sedatives and opioids reduced reports of any pain during surgery by 56% (95% confidence interval [CI], 34%, 70%), but increased nausea and vomiting (odds ratio, 2.27; 95% CI, 1.26, 4.09) compared with those administered topical anesthesia alone, after adjusting for age, gender, race, American Society of Anesthesiologists risk class, self reported health status, and duration of surgery. Among those receiving local injections, use of opioids reduced reports of any pain among those receiving sedatives by 37% (95% CI, 15%, 54%), but did not increase postoperative side effects. The use of diphenhydramine among those receiving sedatives decreased reports of any pain by 59% (95% CI, 33%, 75%) and also reduced drowsiness and nausea and vomiting by 57% (95% CI, 48%, 65%) and by 60% (95% CI, 36%, 75%), respectively. Use of hypnotics with sedatives was associated with increased reports of any pain during surgery and increased nausea and vomiting after surgery. CONCLUSIONS: Patient reports of any pain during cataract surgery (5%) and postoperative side effects (16% drowsiness and 4% nausea and vomiting) were low, but varied by anesthesia strategy. Patient perceptions of pain and side effects can be helpful in guiding the appropriate choice of anesthesia strategy. PMID- 11054332 TI - Corneal surface topography and associated visual performance with INTACS for myopia: phase III clinical trial results. The INTACS Study Group. AB - OBJECTIVE: To characterize corneal topography with INTACS (KeraVision, Inc., Fremont, CA) an ophthalmic device designed to correct myopia, and relate findings to visual performance. DESIGN: Prospective nonrandomized self-controlled comparative intervention study. PARTICIPANTS/INTERVENTION: Patients were participants in the INTACS FDA phase III clinical trials. MAIN OUTCOME MEASURES: Preoperative and postoperative month 6 videokeratographic corneal topography (EyeSys, Houston, TX) was analyzed for 165 eyes from eight clinical sites. Topographic flattening, asphericity, and surface characteristics were statistically evaluated for relationship to visual acuity, refractive data, contrast sensitivity, and subjective visual symptoms. RESULTS: Corneal radius of curvature flattening was aspheric in nature and increased incrementally and significantly for progressively thicker INTACS (P < 0.05). Comparative stratification analyses suggest potential interactions between existing preoperative asphericity and myopia, postoperative asphericity, and visual performance outcomes. Qualitative symmetric and asymmetric toric topography patterns were related to the postoperative self-reported visual symptoms of "double images" (P < or = 0.05) and "halos" (P < or = 0.10), respectively. CONCLUSIONS: The anterior corneal surface is aspherically flattened (prolately) with INTACS, whereas postoperative corneal asphericity is significantly more prolate than preoperative. Specific qualitative postoperative topography patterns were associated with subjective clinical visual performance. PMID- 11054333 TI - Treatment of myopia and myopic astigmatism by customized laser in situ keratomileusis based on corneal topography. AB - OBJECTIVE: To evaluate the predictability, efficacy, and safety of customized laser in situ keratomileusis (LASIK) based on corneal topography in myopia and myopic astigmatism. DESIGN: Prospective, noncomparative interventional case series. PARTICIPANTS: One hundred fourteen patients (eyes) with myopia of -1 to 6 diopters (D) and astigmatism of 0 to -4 D (low myopia group), and 89 patients (eyes) with myopia of -6.10 to -12.00 D and astigmatism of 0 to -4.00 D (high myopia group). INTERVENTION: LASIK was performed with the Hansatome Microkeratome and the Keracor 217 spot-scanning excimer laser (Bausch & Lomb Surgical Technolas, Munich, Germany). Individual ablation patterns were calculated on the basis of elevation data obtained with the Orbscan II corneal topography system (Bausch & Lomb Surgical, Irvine, CA). MAIN OUTCOME MEASURES: Manifest spectacle refraction, visual acuity, and change in visual acuity at 3 months after surgery. RESULTS: At 3 months, 51 patients in the low myopia group and 40 patients in the high myopia group were available. In the low (high) myopia group, 96.1% (75.0%) were within +/-0.50 D of emmetropia, and uncorrected visual acuity was 20/20 or better in 82.4% (62.5%), 20/25 or better in 98.0% (70.0%), and 20/40 or better in 100% (95.0%). A loss of two or more lines of spectacle-corrected visual acuity occurred in 3.9% of the low and 5. 0% of the high myopia group. In low myopia, spectacle-corrected visual acuity was 20/12.5 or better in 5.9% preoperatively and in 13.7% at 3 months and 20/15 or better in 37.3% and 47.1%, respectively. Differences were statistically significant. CONCLUSIONS: The customized LASIK based on corneal topography used in this study showed high predictability and efficacy in myopia and myopic astigmatism of -1.00 to -6.00 D, and could possibly improve spectacle-corrected visual acuity in myopia of -1.00 to -6.00 D. Predictability and efficacy were somewhat lower in myopia and myopic astigmatism of -6.10 to -12.00 D. In both groups, a small number of patients lost two or more lines of spectacle-corrected visual acuity. PMID- 11054334 TI - Discussion by roy S. Rubinfeld, MD PMID- 11054335 TI - Endokeratoplasty as an alternative to penetrating keratoplasty for the surgical treatment of diseased endothelium: initial results. AB - PURPOSE: To test the feasibility of a new surgical technique aimed at replacing diseased corneal endothelium while minimizing visual recovery time. DESIGN: Noncomparative, prospective, clinic-based, interventional case series. PARTICIPANTS: A total of seven patients with aphakic bullous keratopathy (n = 2), pseudophakic bullous keratopathy (n = 4), or Fuchs' corneal dystrophy (n = 1) participated. INTERVENTION: All patients underwent a surgical procedure including removal of the epithelium, creation of a 9.5-mm corneal flap, substitution of an underlying 6.5-mm button of deep stroma and endothelium with a 7.0-mm donor button, and suturing of the flap back into position using a 10-0 running nylon suture. In the two most recently operated patients, anterior lamella, 160 microm in thickness, was removed from the donor button before transplantation. MAIN OUTCOME MEASURES: Visual acuity, refraction, keratometry, corneal topography. RESULTS: All corneas were clear, and the surface reepithelialized within 4 weeks after surgery. Regular astigmatism of less than 4 diopters was recorded in all cases as early as 4 weeks after surgery. Epithelial interface ingrowth with extensive melting of the corneal flap was observed in one patient 3 months after surgery and was managed by removal of the flap and resuturing of the donor button. CONCLUSIONS: Endokeratoplasty represents a promising alternative to conventional penetrating keratoplasty for patients with diseased corneal endothelium. PMID- 11054336 TI - Discussion by herbert E. Kaufman, MD PMID- 11054338 TI - Discussion by peter R. Laibson, MD PMID- 11054337 TI - Primary graft failure : a clinicopathologic and molecular analysis. AB - OBJECTIVE: Primary graft failure (PGF) corneal tissues were analyzed for herpes simplex virus (HSV) and varicella-zoster virus (VZV). DESIGN: Retrospective, noncomparative case series. MATERIALS: Formalin-fixed, paraffin-embedded tissue of 21 donor corneas and 14 recipient corneas of PGF cases, as well as 10 control corneas. METHODS: Clinical, histologic, immunohistochemical, polymerase chain reaction (PCR), and, in selected cases, transmission electron microscopic characteristics were studied. MAIN OUTCOME MEASURES: Evidence of HSV or VZV in donor tissues. RESULTS: Median patient age was 65 years, and median donor age was 48 years. Donor cornea parameters, including endothelial cell counts, death-to preservation time, and time in storage, were generally within accepted standards. Stromal edema was found in all 21 donor corneas with PGF. Eighteen donor corneas demonstrated severely reduced or absent endothelium and mild to moderate lymphocytic infiltration without necrosis. Three donor corneas (14%) had necrotizing stromal keratitis (NSK) with keratic precipitates. Positive immunohistochemical staining of keratocytes for HSV was present in two of two donor corneas with NSK and was negative in 18 other donor corneas. Polymerase chain reaction analysis revealed the DNA of HSV type 1 (HSV1) in all donor corneas with NSK and in four donor corneas without NSK (33%). Recipient corneal tissue was negative for HSV1 DNA in three patients with NSK and positive in two of the four other PCR-positive patients. Transmission electron microscopy analysis showed viral particles in two donor corneas with NSK. Polymerase chain reaction analysis revealed no evidence of HSV type 2 or VZV in any cornea. All control corneas were negative for viral DNA. Sixteen corneas remained clear and two had failed after regraft for PGF, with a median follow-up of 3.6 years. CONCLUSIONS: Herpes simplex virus type 1 DNA was present in 33% of patients of PGF. Herpetic stromal keratitis was found in some failed corneas; the lack of HSV in the paired recipient suggests importation within the donor cornea. The overall prognosis for regrafting after PGF is good. PMID- 11054339 TI - Acute primary angle closure in an Asian population: long-term outcome of the fellow eye after prophylactic laser peripheral iridotomy. AB - PURPOSE: To study the long-term outcome of fellow eyes of Asian patients with acute primary angle closure (APAC) who underwent prophylactic laser peripheral iridotomy (LPI). DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: Ninety-six consecutive patients with APAC at presentation to one Singapore hospital from January 1990 through December 1994. METHODS: The presenting features of the fellow eye were recorded, and the subsequent long-term intraocular pressure (IOP) outcome after LPI was analyzed. All fellow eyes were initially treated with pilocarpine 2% eyedrops four times daily before LPI, which was performed within 1 week of presentation. For any eye, a rise in IOP during follow-up was defined as a rise in IOP requiring treatment by medication or surgery. MAIN OUTCOME MEASURES: Incidence of acute angle closure and IOP. RESULTS: The mean follow-up period was 50.8 months (range, 9-99 months). Of the 96 patients, 15 patients had bilateral APAC, and APAC developed in one fellow eye before LPI could be performed. The remaining 80 fellow eyes were studied. No cases of APAC developed after prophylactic LPI. Seventy-one fellow eyes (88.8%) were successfully treated with LPI alone without the need for additional glaucoma treatment in the long term. Seven eyes (8.8%) had IOPs of 21 mmHg or less on presentation, but a rise in IOP developed on follow-up despite the presence of a patent LPI. Two fellow eyes (2.5%) had signs of preexisting chronic angle closure glaucoma at presentation and required further glaucoma treatment even after LPI. There were no significant complications from the procedure in any of the fellow eyes studied. CONCLUSIONS: In this Asian population with APAC, prophylactic LPI is safe and effective in preventing acute angle closure in fellow eyes. In addition, prophylactic LPI prevents long-term rise in IOP in 88.8% of fellow eyes (with approximately 4 years of follow-up). However, because a small proportion of fellow eyes did experience a rise in IOP within the first year, despite the presence of a patent LPI, close monitoring is still advised in the follow-up of fellow eyes of patients with APAC. PMID- 11054340 TI - Ocular hypertensive response to topical dexamethasone in children: a dose dependent phenomenon. AB - OBJECTIVE: To investigate the ocular-hypertensive response to different dosages of topical dexamethasone eye drops in Chinese children. DESIGN: Prospective, randomized clinical trial. PARTICIPANTS: Thirty-one consecutive children undergoing bilateral strabismus surgery. INTERVENTION: Topical dexamethasone (0.1%) was administered to children undergoing bilateral strabismus surgery. They were all less than 10 years of age. One eye was randomized to receive a regimen of four times daily, and another received a twice daily regimen. Intraocular pressure (IOP) was serially measured in the postoperative period for 4 weeks or more. Topical steroids were discontinued if IOP was 30 mmHg or more. MAIN OUTCOME MEASURES: Intraocular pressure was measured on the day before the surgery, on postoperative days 1, 3, 5, 8, 12, 15, 22, 29, and 2 weeks thereafter until the IOP reached preoperative levels. Peak IOP, IOP net increase, and time to reach an IOP more than 20 mmHg in the two study groups were analyzed. RESULTS: A total of 31 patients (20 male, 11 female) were examined. The mean age was 5.8 +/- 2.0 years (range, 2-10 years). Preoperative IOP in groups treated twice daily and four times daily were similar. After topical dexamethasone treatment, both groups showed a significant rise in peak IOP compared with preoperative values (twice daily, 25.2 +/- 6.8 mmHg vs. 14.3 +/- 2.4 mmHg, P < 0.001; four times daily, 28.7 +/- 6.9 mmHg vs. 14.3 +/- 2.9 mmHg, P < 0.001). The peak IOP was significantly higher in the four times daily group (P < 0.001), as was the net increase in IOP (twice daily, 10.9 +/- 5.8 mmHg vs. four times daily, 14.5 +/- 6.4 mmHg; P < 0.001). There was no difference in time for both groups to achieve the peak IOP, but the time to exceed its upper normal value (20 mmHg) was shorter in the four times daily group (twice daily, 12.3 +/- 9.1 days vs. four times daily, 10.0 +/- 7.4 days; P < 0.05). CONCLUSIONS: In children treated with topical dexamethasone, ocular hypertension occurs in a dose-dependent manner. Children in the four times daily group had a quicker onset and more severe ocular hypertensive response than the twice daily group. Nevertheless, even the twice daily regimen produced significant IOP rise, suggesting that dexamethasone use in children should be avoided if possible, and it would be desirable to monitor the IOP twice weekly when it is administered to children. PMID- 11054341 TI - Nerve fiber analyzer and short-wavelength automated perimetry in glaucoma suspects: a pilot study. AB - PURPOSE: To test the relationship between the results of short-wavelength automatic perimetry (SWAP) and retinal nerve fiber layer (RNFL) measurements with scanning laser polarimetry (Nerve Fiber Analyzer, NFA) in age-matched normal subjects, glaucoma suspects, and early glaucoma patients. DESIGN: Case-control study. PARTICIPANTS AND METHODS: Thirty-eight normal subjects, 32 glaucoma suspects, and 14 early glaucoma patients were recruited. All subjects underwent RNFL assessment by NFA, achromatic visual field testing (24-2 threshold), and repeated SWAP (24-2 threshold blue-on-yellow). MAIN OUTCOME MEASURES: Mean deviation (MD) of visual field testing and RNFL values were obtained. RESULTS: Glaucoma suspects were divided into two groups according to their SWAP results: high risk (with SWAP abnormalities) and low risk (with normal SWAP result). No statistically significant difference in SWAP MD and RNFL values were observed between normal and low-risk groups (P > 0.05), but these values were found to be significantly lower in high-risk and early glaucoma groups (P < 0.01). CONCLUSIONS: This study suggests that RNFL examination by NFA may be a useful test for the early detection of glaucomatous damage of glaucoma suspects. It appears to provide agreement with SWAP abnormalities and is more sensitive than conventional standard automated perimetry. PMID- 11054342 TI - Two-staged Baerveldt glaucoma implant for childhood glaucoma associated with Sturge-Weber syndrome. AB - PURPOSE: To report the outcome and complications of 10 eyes of 9 children with Sturge-Weber syndrome (SWS) who underwent two-stage insertion of a Baerveldt glaucoma implant (BGI) for glaucoma. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: The authors reviewed the medical records of children under the age of 14 years with SWS who underwent two-stage BGI for glaucoma at two tertiary care referral centers. MAIN OUTCOME MEASURES: Intraoperative and postoperative complications, intraocular pressure (IOP), number of glaucoma medications, visual acuity, and further surgical intervention. RESULTS: Ten eyes of nine patients were included in the study. Ages of the nine patients at time of first stage BGI ranged between 6 weeks and 13 years. With average follow-up of 35 months (range, 10-50), all eyes had adequate IOP control (< or = 21 mmHg) without the need for additional glaucoma surgery. Intraocular pressure was reduced from a mean (+/- standard deviation) of 24.8 +/- 6.2 mmHg preoperatively to 16.9 +/- 2.3 mmHg at last follow-up visit (P = 0.001). The number of medications used for control of glaucoma was reduced from a mean (+/- standard deviation) of 1.8 +/- 1.0 preoperatively to 1.1 +/- 1.4 at last follow-up visit (P = 0.2). One eye had serous choroidal effusions with overlying serous retinal detachment that resolved spontaneously after 7 days with no permanent visual loss, and one eye had low choroidal effusion that lasted 4 days. There were no intraoperative or postoperative suprachoroidal hemorrhages. At last follow-up, visual acuity had improved by one or more lines in all patients in whom vision was measurable. CONCLUSIONS: Two-stage BGI surgery appears to be a safe and effective treatment for refractory glaucoma in children with SWS. PMID- 11054343 TI - Pathologic findings in late endophthalmitis after glaucoma filtering surgery. AB - OBJECTIVE: To report the clinicopathologic features of four eyes enucleated for late-onset bleb-related endophthalmitis. STUDY DESIGN: Retrospective case series. MATERIALS: Four enucleated eyes. METHODS: The clinical and histopathologic features of four patients who underwent enucleation for late-onset endophthalmitis after glaucoma filtering surgery were reviewed. RESULTS: The eyes were enucleated for endophthalmitis one to five years after trabeculectomy. Two of the four eyes had trabeculectomy with adjunctive mitomycin-C. All four eyes had streptococci cultured from the aqueous and/or vitreous. Common pathologic features included inflammation involving the anterior segment, lens and choroid. One eye exhibited focal granulomatous uveitis. CONCLUSIONS: Late-onset endophthalmitis after glaucoma filtering surgery is often due to streptococcal species and rapidly progresses over a few days. Phacoanaphylaxis with associated granulomatous uveitis may contribute to the poor prognosis in this setting. PMID- 11054344 TI - Ischemic nephropathy: clinical characteristics and treatment. AB - Ischemic nephropathy is a long-term cause of hypertension and renal failure. Although its real incidence is unknown, ischemic nephropathy is growing because of the increased mean age of the population and the greater prevalence of hypertensive and diabetic populations. This review describes the clinical profile of afflicted patients. Atherosclerosis in different vascular beds is common in these patients. The evolution of ischemic nephropathy is generally progressive, although some patients present with acute renal failure, either secondary to the administration of angiotensin-converting enzyme inhibitors or caused by thrombosis of the renal arteries. Revascularizing surgery may stabilize or improve renal function, even in patients with nonfunctioning kidneys. The results obtained with intraluminal angioplasty are worse, with a high percentage of restenosis. Placement of an endoprothesis is recommended when the lesions affect the ostium or proximal third of the artery. This complex disease typically affects multiple organs, thus making individual assessment essential. PMID- 11054345 TI - ACE inhibitors attenuate expression of renal transforming growth factor-beta1 in humans. AB - Progressive nephropathies are characterized by the enhanced accumulation of extracellular matrix in the kidney. Overproduction of transforming growth factor beta (TGF-beta) was shown to result in pathological tissue fibrosis through the accumulation of extracellular matrix proteins. It has been proposed that angiotensin II stimulates TGF-beta production. Despite accumulating data supporting the effects of angiotensin-converting enzyme (ACE) inhibitors on the attenuation of TGF-beta in vitro and in rats, such studies in humans are lacking. The present study sought to determine the effects of ACE inhibitors on TGF-beta1 in patients with glomerulonephritis. Using competitive polymerase chain reaction and the sandwich enzyme-linked immunosorbent assay, TGF-beta1 messenger RNA (mRNA) abundance and TGF-beta1 protein levels were measured. Patients with immunoglobulin A nephropathy administered ACE inhibitors showed significantly lower renal TGF-beta1 gene expression than patients not administered these medications (mean ratios of TGF-beta1/beta-actin, 4.27 +/- 0.62 [SEM] versus 14.81 +/- 3.87; P < 0.05), whereas no difference was noted between patients administered ACE inhibitors and healthy controls (4.27 +/- 0.62 versus 2.78 +/- 0.71). ACE inhibitor therapy did not affect TGF-beta1 mRNA expression in freshly isolated mononuclear cells. Urine and serum TGF-beta1 protein levels were not affected by the administration of ACE inhibitors. However, possibly a longer duration of treatment would decrease TGF-beta1 levels in urine or blood. In conclusion, we observed a significant reduction in TGF-beta1 expression in the kidney by ACE inhibitors, and this suggests that the effects of ACE inhibitors observed in animals can be extrapolated to patients with chronic renal disease. PMID- 11054346 TI - Risk factors for microalbuminuria in black americans with newly diagnosed type 2 diabetes. AB - We conducted a cross-sectional analysis to describe the prevalence of and risk factors for microalbuminuria among blacks with newly diagnosed type 2 diabetes. Black adults with diagnosed type 2 diabetes mellitus of 2 years' duration or less who presented for care to the Grady Diabetes Clinic (Atlanta, GA) between January 1, 1994, and December 31, 1996, were eligible (n = 1,167). Information obtained at the initial visit included age; sex; body mass index (BMI); serum total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglyceride, C-peptide, serum creatinine, and hemoglobin A1c (HbA(1c)) levels; and seated systolic and diastolic blood pressures. Outcome was urine albumin-creatinine (Alb/Cr) ratio at the initial visit. Alb/Cr ratios were categorized as normal (Alb/Cr <25 microgram/mg), microalbuminuric (Alb/Cr, 25 to 250 microgram/mg), and macroalbuminuric (Alb/Cr >250 microgram/mg). Patients with macroalbuminuria or creatinine levels of 2 mg/dL or greater were excluded. We used multiple linear regression to assess the joint association between HbA(1c) level, mean arterial pressure (MAP), and log-transformed Alb/Cr, controlling for other covariates. Of 1,044 patients studied, macroalbuminuria was present in 3.8%, and microalbuminuria, in 23.4%. Alb/Cr was independently associated with increased HbA(1c) level (P = 0.0070), MAP (P = 0.0001), BMI (P = 0.0156), log transformed triglyceride levels (P = 0.0031), C-peptide level of 6.5 ng/mL or greater (P = 0.0007), serum creatinine level (P: = 0.0068), and male sex (P = 0.0220). The relationship between HbA(1c) level and microalbuminuria was stronger in patients with lower BMIs. Microalbuminuria prevalence was high in this population of urban blacks with newly diagnosed type 2 diabetes. Risk factors associated with increased Alb/Cr included male sex, poor glycemic control, endogenous hyperinsulinemia, high blood pressure, elevated triglyceride levels, and obesity. PMID- 11054347 TI - Pauci-immune renal vasculitis: natural history, prognostic factors, and impact of therapy. AB - The purpose of this study is to describe the clinical presentation and natural history of pauci-immune renal vasculitis and determine whether particular presenting features or administered therapies predict outcome. We reviewed our experience since 1984 with such vasculitides, and 94 cases of pauci-immune vasculitis were identified. Presenting features were as follows: men, 63%; mean age at biopsy, 59 years; and mean serum creatinine level, 5.0 mg/dL. Patients with no extrarenal involvement had a tendency to present with a greater serum creatinine level. Since the antineutrophil cytoplasmic autoantibody (ANCA) assay became available, 77% of the patients tested were ANCA positive. In terms of natural history, 27 patients required dialysis immediately, there were 22 renal relapses, 28 patients progressed to dialysis, 10 patients died before requiring dialysis, 19 patients were lost to follow-up, and 37 patients remain active, not on dialysis. Overall, half the patients recovered some renal function, one third remained stable, and one sixth deteriorated. Female sex and angiotensin converting enzyme inhibitor use predicted favorable outcome (P < 0.05). Advanced age, male sex, respiratory tract involvement, and a greater relapse rate predicted unfavorable outcome (P < 0.05). The incidence of pauci-immune renal vasculitis appears to be increasing, likely related to the emergence of the ANCA assay. Attempts to classify patients based on existing schemes may result in delayed diagnosis and therapy, with subsequent poorer outcomes. Also, given the increased mortality of patients with respiratory tract involvement, we speculate that respiratory tract disease therapeutic and monitoring regimens are ineffective. In general, we conclude that pauci-immune renal vasculitis is a heterogeneous disorder with an unfavorable prognosis. PMID- 11054348 TI - A C677T mutation in the methylenetetrahydrofolate reductase gene modifies serum cysteine in dialysis patients. AB - Patients undergoing hemodialysis have impaired metabolism of such sulfur containing amino acids as cysteine (Cys) and homocysteine (Hcy), which may lead to accelerated atherosclerosis. Considering that Cys is mainly synthesized from Hcy, a common C677T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene may affect the serum total Cys (tCys) concentration, as well as total Hcy (tHcy) concentration, through reduced remethylation of Hcy to methionine, even in hemodialysis patients. To identify the independent determinants for the tCys concentration in dialysis patients, we determined MTHFR C/T genotypes and serum concentrations of tHcy, tCys, and vitamins as cofactors in 464 hemodialysis patients. Serum tCys concentration was positively associated with serum tHcy concentration and negatively associated with the MTHFR mutation, although the mutation correlated positively with serum tHcy concentration. Slopes of regression lines relating tHcy and tCys concentrations differed between the MTHFR genotypes, and the relationship was strengthened with a decreasing number of T alleles. Additionally, serum concentrations of folate and vitamin B(12) correlated positively with tCys concentration, whereas they correlated negatively with tHcy concentration. These findings suggest that the MTHFR mutation is an independent predictor for serum tCys concentrations in hemodialysis patients and that a tCys-decreasing effect of the mutation may arise largely from its attenuation of the positive Cys-Hcy correlation. The tCys-increasing effect of folate and vitamin B(12) appears to be linked to their enhancement of Hcy remethylation. PMID- 11054349 TI - 8-hydroxy-2'-deoxyguanosine of leukocyte DNA as a marker of oxidative stress in chronic hemodialysis patients. AB - In contrast to proteins and lipids, oxidative damage to DNA has not been well studied in patients undergoing hemodialysis (HD). We hypothesized that phagocytes are activated after blood-membrane contact during HD, and oxidants from metabolic activation can damage leukocyte DNA. To test this hypothesis, the 8-hydroxy-2' deoxyguanosine (8-OHdG) content of leukocyte DNA was measured by high-performance liquid chromatography electrochemical detection method in 35 age- and sex-matched healthy subjects, 22 undialyzed patients with advanced renal failure, and 109 HD patients to assess the relation between oxidative DNA damage and complement activating membranes, blood antioxidants, and iron status. Dialysis membranes were classified into complement-activating (cellulose; n = 55) and non-complement activating (polymethylmethacrylate [PMMA]; n = 35; polysulfone [PS]; n = 19) membranes. We found increased oxidative stress in undialyzed and HD patients based on a decrease in plasma levels of ascorbate and alpha-tocopherol adjusted for blood lipid (alpha-tocopherol/lipid), serum albumin, and reduced glutathione levels in whole blood and an increase in oxidized glutathione levels in whole blood compared with controls (P < 0.001). The greatest 8-OHdG level in leukocyte DNA was in HD patients, followed by undialyzed patients and healthy controls (P < 0.001), and was significantly greater in HD patients using cellulose membranes than those using PMMA or PS membranes (P < 0.001). 8-OHdG levels correlated with plasma alpha-tocopherol/lipid (r = -0.314; P < 0.005), serum iron (r = 0. 446; P < 0.001), and transferrin saturation values (r = 0.202; P < 0.05) in the analysis of all HD patients. In a 6-week crossover study, 8-OHdG levels significantly decreased after the switch from cellulose to synthetic membranes for 2 weeks and increased after the shift from synthetic to cellulose membranes (P < 0.05). Iron metabolism indices and plasma alpha-tocopherol/lipid values did not change significantly in the study period. We conclude that 8-OHdG content in leukocyte DNA is a biomarker of oxidant-induced DNA damage in HD patients. Oxidative DNA damage is a consequence of uremia, further augmented by complement-activating membranes. PMID- 11054350 TI - Hepatocyte growth factor predicts survival and relates to inflammation and intima media thickness in end-stage renal disease. AB - Hepatocyte growth factor (HGF) is a pleiotropic cytokine involved in tissue protection and repair in the endothelium and various organ systems. The serum concentration of this protein is markedly increased in patients with chronic renal diseases, but the clinical and pathophysiological correlates of this substance in renal failure are scarcely understood. Serum HGF, lipid, albumin, hemoglobin, C-reactive protein (CRP), and immunoglobulin G (IgG) were measured in fasting conditions in a cohort of 244 dialysis patients. In addition, the relationship between HGF and severity of carotid atherosclerosis was studied in a subgroup of 105 patients. The entire cohort was followed up for a median of 31 months (interquartile range, 21 to 34 months). Serum HGF level was directly related to duration of dialysis treatment, CRP level, age, IgG level, and hemoglobin level and inversely related to systolic and diastolic arterial blood pressure. In a multiple regression model, only duration of dialysis treatment (r = 0.38), age (r = 0.26), hemoglobin level (r = 0.17), IgG level (r = 0.15), and CRP level (r = 0.14) were independent correlates of serum HGF level (R = 0.54; P < 0.0001), suggesting that increased levels of serum HGF may be the expression of a chronic inflammatory process. HGF levels were greater in hemodialysis than continuous ambulatory peritoneal dialysis patients, independent of the type of dialysis membrane, and slightly increased in patients seropositive for hepatitis C virus. In the subgroup of patients who underwent echo color Doppler studies, serum HGF level was an independent correlate of intima media thickness (IMT; partial r = 0.23; P = 0.02). In the entire cohort, increased HGF levels predicted shorter survival in a multivariate Cox regression model. These results support the hypothesis that in patients with chronic renal failure, increased serum HGF level is linked to an inflammatory state. The relationships between HGF level and survival and IMT suggest that this cytokine might be a marker of a process that has a major impact in the high mortality and morbidity of the dialysis population. PMID- 11054351 TI - Parathormone secretion in peritoneal dialysis patients with adynamic bone disease. AB - The prevalence of low-turnover lesions in patients undergoing peritoneal dialysis (PD) is high. Our aims are to evaluate the prevalence of adynamic bone disease (ABD) in PD patients, analyze risk factors, and define the association of serum parathyroid hormone (PTH) levels measured under different plasma calcium concentrations with this lesion. Fifty-seven patients were studied by bone biopsy (BB). ABD was found in 63.2%, and 36.8% showed high-turnover bone disease (HTBD). Patients with HTBD had a lower prevalence of diabetes, younger age, lower accumulated oral calcium salt intake, and greater calcitriol doses, serum osteocalcin level, and ultrafiltration than patients with ABD. Both mean baseline PTH levels from the previous year and PTH level at time of BB were greater in patients with HTBD than those with ABD (357 +/- 267 pg/mL versus 89 +/- 67 pg/mL; 390 +/- 337 pg/mL versus 88 +/- 78 pg/mL, respectively; P < 0.05). However, the magnitude of the increase from baseline serum PTH levels in response to hypocalcemia was greater in patients with ABD than in those with HTBD (166.4% +/- 134% versus 83.5% +/- 73.6%; P < 0.05). We found that PTH levels less than 150 pg/mL in patients with ABD showed a sensitivity of 91. 6%, specificity of 95.2%, and positive predictive value (PPV) of 97%. In the HTBD group, PTH levels greater than 450 pg/mL had a specificity and PPV of 100%. Our data confirm that ABD is the most prevalent lesion in PD patients, and PTH secretion capacity is maintained in these patients. The definitive diagnosis and management strategies for many patients requires a BB, especially when HTBD is unlikely. PMID- 11054352 TI - Delayed gastric emptying in dyspeptic chronic hemodialysis patients. AB - Hemodialysis patients frequently experience such dyspeptic symptoms as nausea, vomiting, abdominal distension, early satiety, and anorexia. Gastroparesis might be a cause of malnutrition, and parameters of gastric emptying are inversely correlated with serum albumin levels. The aim of the present study is to determine whether delayed gastric emptying is related to dyspeptic symptoms. In 54 hemodialysis patients, a standardized history for dyspeptic symptoms was taken. In addition, gastric emptying for solids was measured in 26 patients, using the (13)C-octanoic acid breath test. There was a high prevalence of dysmotility-like dyspepsia in the hemodialyzed population. A significant difference in gastric emptying between dyspeptic hemodialysis patients and healthy volunteers and between dyspeptic and nondyspeptic hemodialysis patients was shown. There was a significant correlation between gastric emptying and dysmotility-like dyspepsia. Serum albumin level inversely correlated with gastric emptying. In conclusion, there is a high prevalence of dysmotility-like dyspepsia in hemodialysis patients. Dyspeptic patients have significantly delayed gastric emptying compared with both healthy volunteers and nondyspeptic patients. PMID- 11054353 TI - Use of ultrafiltration and chromatography to assess aluminum speciation in serum after deferoxamine administration. AB - Deferoxamine effectively chelates aluminum by forming aluminoxamine, a low molecular-weight compound removable by dialysis. However, aluminum-bound species other than aluminoxamine might be present in serum after the administration of deferoxamine. To study aluminum speciation after the administration of deferoxamine, high-performance liquid chromatography (HPLC) and ultrafiltration techniques were used. Samples of serum were obtained from six dialysis patients 44 hours after the administration of a single dose of deferoxamine. HPLC and ultrafiltration studies were performed. In the HPLC studies, samples underwent ultrafiltration, the filtrate was injected into the chromatographic system, and detection was performed by UV light and atomic absorption spectrometry. Unknown species of aluminum other than aluminoxamine were found in the early elution fractions. In the ultrafiltration studies, the same samples of serum from the six patients underwent ultrafiltration using membranes with different molecular weight cutoff values from 1 to 30 kd. The percentages of aluminum found by ultrafiltration using membranes with cutoff values of 5, 10, and 30 kd were greater (64.4% +/- 2.5%, 63.5% +/- 3.7%, and 65.6% +/- 4.3%, respectively) than the percentages obtained with membranes with a 1-kd cutoff value (38.7%), suggesting that the unknown species of aluminum have a molecular weight between 1 and 5 kd. The unknown species of aluminum cannot be aluminoxamine because they behaved in a different way with HPLC. PMID- 11054354 TI - A randomized comparison of intradermal and intramuscular vaccination against hepatitis B virus in incident chronic hemodialysis patients. AB - Hepatitis B virus (HBV) remains a threat to hemodialysis patients. Nevertheless, the vaccination rate against this virus is low, perhaps because of the low conversion rate. Although intradermal (ID) vaccination has been proven to be effective (even in patients nonresponsive to intramuscular [IM] vaccination), the duration of immunity was short. The impact of vaccination route and a greater peak antibody (Ab) titer on conversion rate and duration of immunity after ID or IM vaccination was compared in incident hemodialysis patients. Forty-nine patients were randomly assigned to treatment with 5 microgram of recombinant hepatitis B vaccine (Engerix B; Smith Kline Beecham Pharma Inc, Oakville, ON, Canada) ID every 2 weeks up to either a peak Ab titer of 1,000 IU/L or greater or 52 doses, whereas 48 patients were administered 40 microgram IM at 0, 1, 2, and 6 months. Group demographics were similar. Conversion was achieved in 97.6% of the ID group and 90.5% of the IM group (P: = 0.16). There was no difference between ID and IM groups in time required to convert, peak Ab titer reached, and proportion of patients with a peak Ab titer of 1,000 IU/L or greater. Overall, the duration of immunity was not different after ID or IM vaccination (P = 0.683), and patients in the IM group with a peak Ab titer of 1,000 IU/L or greater had a longer duration of immunity (P = 0.001). In conclusion, a high conversion rate and long duration of immunity against HBV can be achieved cost effectively in the end-stage renal disease population using the ID or IM route and aiming for an Ab titer exceeding 1,000 IU/L. Based on these data, we provide recommendations for vaccination and surveillance in this population. PMID- 11054355 TI - Reproducibility of ambulatory blood pressure monitoring in hemodialysis patients. AB - Ambulatory blood pressure monitoring (ABPM) has been increasingly used in hemodialysis (HD) practice and research; however, no study has evaluated the reproducibility of ABPM in this population. To address this question, we performed 48-hour interdialytic ABPM on 21 HD patients (mean age, 53 +/- 16 years; 7 women) on two different occasions 68 +/- 34 days (range, 30 to 154 days) apart. To qualify for the protocol, patients had to be at the same dry weight and on the same vasoactive drug regimen at both monitoring periods. BP was analyzed according to three different methods: isolated pre-HD and post-HD values, average pre-HD and post-HD values for the five HD sessions surrounding each monitoring period, and 48-hour interdialytic ABPM. Reproducibility was determined by analysis of the SD of the differences (SDD) between the two monitoring periods and the coefficient of variation of each method of BP determination. Our results show better reproducibility of ABPM (SDD, 10.6/6.6 mm Hg; coefficient of variation, 7.5%/8.1%) compared with isolated pre-HD BP (SDD, 24.4/11.3 mm Hg; coefficient of variation, 16.7%/14.1%) or post-HD BP (SDD, 16.8/14.5 mm Hg; coefficient of variation, 11.7%/17.8%), and averaged pre-HD BP (SDD, 14.7/7.2 mm Hg; coefficient of variation, 10.1%/9.1%) or post-HD BP (SDD, 12.4/8.7 mm Hg; coefficient of variation, 8.9%/11.1%). The reproducibility of the decrease in BP during sleep was poor, with up to 43% of the subjects changing dipping category within or between interdialytic periods. We conclude that ABPM is the most accurate method to study BP in HD patients over time. However, variability is significant, and there is poor reproducibility of the nocturnal decline in BP. PMID- 11054356 TI - Characteristics and treatment of patients not reusing dialyzers in reuse units. AB - Dialyzer reuse is practiced in more than 75% of the patients and dialysis units in the United States. However, reuse is not practiced in a small fraction of patients treated in reuse units (RUUs). This study evaluates both patient and facility characteristics associated with nonreuse in RUUs. The data source is from the Dialysis Mortality and Morbidity Study, Waves 1, 3, and 4, of the US Renal Data System. Only facilities that practiced dialyzer reuse were included in the analysis. A total of 12,094 patients from 1,095 reuse facilities were studied. Patients undergoing hemodialysis as of December 31, 1993, were selected. Of all patients treated in RUUs, 8% did not reuse dialyzers. Nonreuse was significantly (P < 0.02) more common, based on adjusted odds ratios (ORs), among patients who were younger (OR = 1.16 per 10 years younger), had primary glomerulonephritis (OR = 1.26 versus diabetes), had lower serum albumin level (OR = 1.72 per 1 g/dL lower), had more years on dialysis, and had higher level of education. Nonreuse patients were more likely to be treated with low-flux dialyzers (OR = 7.35; P < 0. 0001) and have a lower dialysis dose. No reuse was more likely in larger units and in not-for-profit and hospital-based units. Patient refusal accounted for one fourth of nonreuse in RUUs and was associated with the same factors, as well as with fewer comorbid conditions and non-Hispanic ethnicity. Significant geographic variations (up to eightfold) were documented. Nonreuse patients are treated with smaller, low-flux dialyzers and, on average, receive a lower Kt/V than reuse patients in the same units. PMID- 11054357 TI - Polymicrobial peritonitis in continuous ambulatory peritoneal dialysis patients. AB - We retrospectively evaluated 232 continuous ambulatory peritoneal dialysis (CAPD) patients entering our program from January 1, 1987, to December 31, 1997, for polymicrobial peritonitis. Polymicrobial peritonitis occurred in 16% of the patients (polymicrobial-peritonitis group), whereas 52% of the patients had peritonitis episodes with only a single organism (single-organism group), and 32% of the patients had no episode of peritonitis. Polymicrobial peritonitis accounted for 8% of the 554 peritonitis episodes, occurred after 23 +/- 20 months on peritoneal dialysis (PD), and was preceded by greater than three episodes of peritonitis in 73% of the patients. Peritonitis rates were greater in the polymicrobial-peritonitis group compared with patients in the single-organism group (1.8 versus 1.2 episodes/patient-year; P: < 0.001). The majority of polymicrobial infections involved gram-negative and/or fungal pathogens, but in 21% of the episodes, only gram-positive organisms were identified. An intra abdominal process was identified in only 7% of the patients. Catheter loss overall was greatest in the polymicrobial-peritonitis group (65% versus single organism group, 30% versus patients without peritonitis, 5%; P < 0.001), but only 33% of the polymicrobial infections resulted in catheter loss. At last follow-up, 70% of the patients in the polymicrobial-peritonitis group had permanently transferred to hemodialysis compared with 25% from the single-organism group and 15% from the no-peritonitis group (P < 0.001). In conclusion, polymicrobial peritonitis is an infrequent but serious complication of CAPD that occurs late in the course of PD and is often preceded by recurrent episodes of peritonitis. Polymicrobial peritonitis is rarely the result of a catastrophic intra-abdominal process, and although the majority of patients can be successfully treated without catheter removal, the long-term prognosis is poor, with a high rate of transfer to hemodialysis. PMID- 11054358 TI - Analysis of microbiological trends in peritoneal dialysis-related peritonitis from 1991 to 1998. AB - The microbial cause of peritoneal dialysis-related peritonitis is an important determinant of clinical outcome and the basis of widely used treatment guidelines. Five hundred forty-six cases of peritonitis in 374 patients from 1991 to 1998 were analyzed. The rate of peritonitis declined significantly from 1.37 episodes/patient-year in 1991 to 0.55 episode/patient-year in 1998 (P = 0.02). The rate of Gram-positive peritonitis decreased significantly from 0.75 to 0.28 episode/patient-year during the same period (P = 0.02). Conversely, the occurrence of Gram-negative peritonitis remained constant at approximately 0.16 episode/patient-year (P = 0.28). Staphylococcus epidermidis and Staphylococcus aureus were the most common causes of peritonitis, isolated in 27.8% and 19.3% of the culture-positive cases, respectively. A distinct decrease in peritonitis caused by S epidermidis was observed, with 0.40 episode/patient-year in 1991 compared with 0.11 to 0.20 episode/patient-year during subsequent years. The rate of infections caused by S aureus decreased significantly over time from a high of 0.21 episode/patient-year in 1992 to a low of 0.04 episode/patient-year in 1998 (P = 0.01). Pseudomonas aeruginosa, Escherichia coli, and KLEBSIELLA: species were the most common causes of Gram-negative peritonitis, identified in 7.1%, 6.8%, and 5.2% of culture-positive cases, respectively. The most dramatic increase in antibiotic resistance was seen among S epidermidis. From 1991 and 1992 to 1997 and 1998, resistance to ciprofloxacin increased from 5.4% to 47.8% (P = 0.003), and resistance to methicillin increased from 18.9% to 73.9% (P = 0.03). Our study showed significant trends in the causative pathogens of peritoneal dialysis-related peritonitis and dramatic increases in antibiotic resistance. These data support further study and warrant reevaluation of current treatment practices. PMID- 11054359 TI - Role of preoperative antibiotic prophylaxis in preventing postoperative peritonitis in newly placed peritoneal dialysis catheters. AB - The role of vancomycin and other antibiotics in the treatment of acute peritonitis in peritoneal dialysis (PD) patients is well established. However, the role of preoperative vancomycin or cephalosporins in preventing early infection in newly placed PD catheters remains controversial. We performed a prospective randomized study to examine the role of vancomycin or cefazolin prophylaxis in decreasing the incidence of postoperative peritonitis. Over a 6 year period, 221 patients undergoing 254 permanent peritoneal catheter placement procedures were randomized into three groups. Group I patients (86 procedures) were administered a single intravenous (IV) dose of vancomycin, 1,000 mg, 12 hours before peritoneal catheter placement procedures, whereas group II patients (85 procedures) were administered a single IV dose of cefazolin, 1,000 mg, 3 hours before the procedure. Group III patients (83 procedures) were not administered antibiotics preoperatively for at least 1 week before the procedure. Patients were monitored for peritonitis in the following 14 days. Peritonitis developed in 1 patient (1%) in group I (vancomycin group) and 6 patients (7%) in group II (cefazolin group) compared with 10 patients (12%) in group III (control group); P = 0.02. We conclude that the use of preoperative single-dose IV vancomycin prophylaxis for permanent PD catheter placement reduces the risk for postoperative peritonitis. Single-dose vancomycin is superior to single-dose cefazolin in reducing the risk for postoperative peritonitis. Absence of prophylaxis is associated with a high risk for developing postoperative peritonitis. PMID- 11054360 TI - Relationship between phosphorus and creatinine clearance in peritoneal dialysis: clinical implications. AB - Elevated serum phosphorus levels are associated with increased morbidity and mortality in dialysis patients. To study the determinants of serum phosphorus levels and phosphorus clearance, we measured phosphorus concentrations in 24-hour collections of dialysate and residual urine output during routine urea kinetics in 56 peritoneal dialysis patients. Dietary records were used to estimate oral phosphorus intake, and the net gastrointestinal absorption of phosphorus was determined measuring the amount of phosphorus excreted in 24 hours. Dialysate to plasma ratios of phosphorus and creatinine were very similar, and the peritoneal clearances of phosphorus and creatinine correlated well and were of the same magnitude. High transporters in the peritoneal equilibration test had greater phosphorus and creatinine clearances than low transporters. In patients with residual urine output, the renal clearances of phosphorus and creatinine also correlated well. Comparing two groups of patients with a similar Kt/V of 2 and greater but different levels of creatinine clearance, either greater or less than 60 L/wk, the patients with the greater creatinine clearance also had a greater phosphorus clearance (7.0 +/- 2.2 versus 4.3 +/- 0.7 mL/min/1.73 m(2)) and significantly lower serum phosphorus levels (4.8 +/- 0.9 versus 5.9 +/- 1.4 mg/dL) despite a greater daily amount of phosphorus absorption (490 +/- 190 versus 336 +/- 100 mg/d). In summary, creatinine clearance measurements provide a good estimate of phosphorus clearance and how much dietary phosphorus and therefore protein can be tolerated. PMID- 11054361 TI - Differences in access to cadaveric renal transplantation in the United States. AB - This national study compares waitlisting and transplantation rates by gender, race, and diabetes and evaluates physiologic factors (panel-reactive antibodies [PRA], blood type, HLA matchability) and related practices (early and multiple waitlisting) as explanatory factors. This longitudinal study of the time to transplant waitlisting among 228,552 incident end-stage renal disease (ESRD) dialysis patients and to cadaveric transplantation among 46,164 waitlist dialysis patients (n = 23,275 first cadaveric transplants) used US data for 1991 to 1997. Relative rates of waitlisting (RRWL) after ESRD onset and of cadaveric transplantation (RRTx) after waitlist (Cox proportional hazards models) were adjusted for age, race, sex, ESRD cause, region, and incidence/waitlist year. We found that women have an RRWL = 0.84 (P < 0.0001) and RRTx = 0.86 (P < 0. 0001). PRA levels can explain the difference in the transplantation rate, because accounting for PRA gives an adjusted RRTx = 0.98 (NS) for women. For blacks versus whites, the RRWL = 0.59 (P < 0.0001) and RRTx = 0.55 (P < 0.0001). However, the transplantation rate can only partly be explained by ABO types, rare HLA types, and early and multiple waitlisting (adjusted RRTx = 0.67 [P < 0.0001]). For diabetes versus glomerulonephritis, the RRWL = 0.52 (P < 0.0001) and RRTx = 0.98 (NS). Older patients (40 to 59 years of age) are less likely to be waitlisted and to receive a transplant after waitlisting (RRWL = 0.57 [P < 0.0001], RRTx = 0.88 [P < 0.0001]) versus younger patients (ages 18 to 39 years). These results indicate substantial differences by age, sex, race, and diabetes in rates of waitlisting for transplantation and by age and race for transplantation after waitlisting. These differences by race were not explained by referral practices or the physiologic factors studied here. PMID- 11054362 TI - Acute granulomatous interstitial nephritis and colitis in anticonvulsant hypersensitivity syndrome associated with lamotrigine treatment. AB - We present the case of a 17-year-old woman with a history of bipolar disorder, who developed a clinical syndrome manifested by fever, lymphadenopathy, skin rash, diarrhea, and acute renal failure requiring dialysis after the use of lamotrigine. Renal biopsy showed acute interstitial nephritis (AIN) with focal granulomas. Similarly, colonic biopsy specimens showed colitis and ileitis with non-necrotizing epithelioid granulomas. The patient had a complete recovery after withdrawal of the medication and steroid treatment. Although lamotrigine has been previously implicated in the development of anticonvulsant hypersensitivity syndrome, there have been no previous reports of acute granulomatous interstitial nephritis or colitis associated with the use of this drug. PMID- 11054363 TI - Facet joint osteomyelitis in a patient on long-term hemodialysis. AB - Spinal osteomyelitis is a recognized complication in dialysis patients related to hematogenous spread of infection during bacteremia-septicemia. These episodes are often associated with sepsis due to temporary dialysis access. We describe the case of an unfortunate man whose osteomyelitis was located in the posterior facet joints. Such infection is rare and in the reviewed literature is usually associated with a more favorable outcome than described here. PMID- 11054364 TI - Lumbosacral plexopathy after dual kidney transplantation. AB - A 58-year-old man underwent dual kidney transplantation. He was unable to move his right leg after surgery. This was caused by extensive lumbosacral plexopathy on the side of surgery. Lumbosacral plexopathy after kidney transplantation is uncommon, because the plexus has rich anastomotic blood supply, and ischemic injury is unlikely. However, isolated femoral neuropathy after renal transplantation has been reported, as the distal portion of this nerve is supplied by branches of internal iliac artery only and is more prone to ischemic injury during surgery. Dual-kidney transplantation involves a larger dissection, and the procedure takes 60 to 90 minutes longer than single-kidney transplantation. It involves more vascular reconstruction. This may predispose the lumbosacral plexus to ischemic injury. To the best of our knowledge, this is the first reported case of lumbosacral plexopathy after a dual kidney transplantation, and this may be seen more frequently because this procedure is becoming more common. PMID- 11054365 TI - Factitious hyperkalemia. AB - Pseudohyperkalemia, or factitious hyperkalemia, constitutes an artificially high plasma potassium level (P(K)) from a variety of possible causes. Occasionally, the cause cannot be elucidated. Three patients who showed unusually large differences between free-flowing and tourniquet (stasis) potassium levels prompted us to investigate the influence of tourniquets in routine phlebotomy in eight healthy volunteers. P(K) showed a consistent but rather small average increase of 0.2 mEq/L (P < 0.001) during tourniquet use; however, the range was 10-fold, from 0.05 to 0.5 mEq/L in our subjects. We suggest there may be large variability leading to an excessive increase in P(K) in some individuals. In the three patients presented, average excessive increases in P(K) of 1.6, 1.3, and 1.7 mEq/L were seen. Although diagnosing and treating true hyperkalemia remains paramount, recognizing factitious hyperkalemia is important to preclude unnecessary investigations and potentially hazardous intervention. PMID- 11054366 TI - Can risk factor modification prevent nephropathy in type 2 diabetes mellitus? PMID- 11054367 TI - Changing picture of peritonitis in peritoneal dialysis. PMID- 11054368 TI - Rationale for an International Federation of Kidney Foundations. AB - Kidney foundations serve a unique role with their focus on the medical, psychosocial, and health needs of the individual with kidney disease and with their community-based structure. The types of activities in which kidney foundations may become involved include, but are not limited to, educational programs for health care workers and for patients and their families; educational, rehabilitative, and financial support programs for patients or their families; advocacy to the government and to other organizations on behalf of the needs of the patient and the patient's family; and fund raising for research or for other kidney-related programs. Most of the world's population is not represented by kidney foundations. Moreover, there are major variations in the programmatic activities of most kidney foundations. The fact that the psychosocial and educational needs of individuals with renal disease and renal failure are often great and the fact that access of individuals with end-stage renal disease to long-term dialysis therapy or renal transplantation varies greatly in different parts of the world provide a strong rationale for the establishment of community-based kidney foundations to advocate for the patient. The International Federation of Kidney Foundations (IFKF), which was formed during the past year, has as its goal fostering international collaboration and exchange of ideas to improve the health, well-being, and quality of life for individuals with kidney disease. The IFKF will promote the establishment of kidney foundations in regions where none currently exist and will encourage the growth in programmatic activities of kidney foundations everywhere. The increasing globalization of the world, growing affluence worldwide, and the willingness of people to engage in charitable giving suggest that this is a most opportune time to launch this international organization. PMID- 11054369 TI - Angiotensin II subtype-2 receptor influences kininogenase activity PMID- 11054370 TI - Acute renal failure in an HIV-positive 50-year-old man. PMID- 11054371 TI - Impact of K(+) homeostasis on net acid secretion in rat terminal inner medullary collecting duct: role of the Na,K-ATPase. AB - For the past 50 years, the mechanism of ammonium (NH(4)(+)) transport along the collecting duct has been thought to occur through active H(+) section in parallel with the nonionic diffusion of ammonia (NH(3)). This model is supported by two basic experimental observations. First, NH(4)(+) secretion generally correlates with the NH(3) concentration gradient between the interstitium and the collecting duct lumen. This NH(3) gradient is generated through both luminal acidification, which reduces luminal NH(3) concentration, and through countercurrent multiplication, which increases interstitial NH(3) concentration. The result is secretion of NH(3) into the collecting duct lumen down its concentration gradient. Second, because NH(4)(+) permeability is low relative to that of NH(3), there is significant secretion of NH(3) into the collecting duct lumen with minimal back-diffusion of NH(4)(+). However, our laboratory, as well as others, has shown that this model is an oversimplification of the mechanism of NH(4)(+) transport along the collecting duct. NH(4)(+) is transported through a variety of K(+) transport pathways including Na,K-ATPase. K(+) and NH(4)(+) compete for a common extracellular binding site on Na, K-ATPase. During hypokalemia, interstitial K(+) concentration is reduced, which augments NH(4)(+) uptake by the Na(+) pump. In K(+) restriction, Na,K-ATPase-mediated NH(4)(+) uptake provides an important source of H(+) for net acid secretion and for the titration of luminal buffers in the terminal inner medullary collecting duct. This pathway contributes to the increase in NH(4)(+) excretion and metabolic alkalosis observed during hypokalemia. PMID- 11054372 TI - Oxalosis PMID- 11054373 TI - Continuing medical education exercise, november 2000 PMID- 11054374 TI - Hemolytic uremic syndrome: past and present PMID- 11054375 TI - Alternative therapy si, gene therapy no? PMID- 11054376 TI - Primary gastric B-cell lymphoma: results of a prospective multicenter study. The German-Austrian Gastrointestinal Lymphoma Study Group. AB - BACKGROUND & AIMS: Appropriate management of primary gastric lymphoma is controversial. This prospective, multicenter study aimed to evaluate the accuracy of endoscopic biopsy diagnosis and clinical staging procedures and assess a treatment strategy based on Helicobacter pylori status and tumor stage and grade. METHODS: Of 266 patients with primary gastric B-cell lymphoma, 236 with stages EI (n = 151) or EII (n = 85) were included in an intention-to-treat analysis. Patients with H. pylori-positive stage EI low-grade lymphoma underwent eradication therapy. Nonresponders and patients with stage EII low-grade lymphoma underwent gastric surgery. Depending on the residual tumor status and predefined risk factors, patients received either radiotherapy or no further treatment. Patients with high-grade lymphoma underwent surgery and chemotherapy at stages EI/EII, complemented by radiation in case of incomplete resection. RESULTS: Endoscopic-bioptic typing and grading and clinical staging were accurate to 73% and 70%, respectively, based on the histopathology of resected specimens. The overall 2-year survival rates for low-grade lymphoma did not differ in the risk adjusted treatment groups, ranging from 89% to 96%. In high-grade lymphoma, patients with complete resection or microscopic tumor residuals had significantly better survival rates (88% for EI and 83% for EII) than those with macroscopic tumor residues (53%; P < 0.001). CONCLUSIONS: There is a considerable need for improvement in clinical diagnostic and staging procedures, especially with a view toward nonsurgical treatment. With the exception of eradication therapy in H. pylori-positive low-grade lymphoma of stage EI and the subgroup of locally advanced high-grade lymphoma, resection remains the treatment of choice. However, because there is an increasing trend toward stomach-conserving therapy, a randomized trial comparing cure of disease and quality of life with surgical and conservative treatment is needed. PMID- 11054377 TI - Bone mineral density in patients with recently diagnosed inflammatory bowel disease. AB - BACKGROUND & AIMS: A high prevalence of osteoporosis is reported in inflammatory bowel disease (IBD), and its pathogenesis is not completely resolved. We investigated whether bone mineral density (BMD) in patients with IBD at diagnosis is lower than in population controls, and whether BMD differs between patients with Crohn's disease and those with ulcerative colitis. METHODS: In 68 patients and 68 age- and gender-matched population controls, BMD of total body, spine, and hip was assessed using dual-energy x-ray absorptiometry within 6 months after establishing the diagnosis. Determinants for low BMD were assessed. RESULTS: There were no significant differences in BMD (g/cm(2)) between patients and controls, and no significant differences in BMD between patients with either Crohn's disease or ulcerative colitis. Multivariate regression analysis showed that duration of complaints longer than 6 months before diagnosis (P = 0.041), age (P = 0.019), and body mass index less than 20 kg/m(2) (P = 0.006) significantly correlated with low BMD. CONCLUSIONS: BMD in patients with recently diagnosed IBD was not significantly decreased compared with population controls. Subsequent development of osteoporosis in patients with IBD seems to be a phenomenon related to the disease process and/or the treatment modalities of IBD. PMID- 11054378 TI - Suppression of NF-kappaB activity by sulfasalazine is mediated by direct inhibition of IkappaB kinases alpha and beta. AB - BACKGROUND & AIMS: Activation of NF-kappaB/Rel has been implicated in the pathogenesis of inflammatory bowel disease (IBD). Various drugs used in the treatment of IBD, such as glucocorticoids, 5-aminosalicylic acid, and sulfasalazine, interfere with NF-kappaB/Rel signaling. The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-kappaB activation. METHODS: The effects of sulfasalazine and its moieties on NF-kappaB signaling were evaluated using electromobility shift, transfection, and immune complex kinase assays. The direct effect of sulfasalazine on IkappaB kinase (IKK) activity was investigated using purified recombinant IKK-alpha and -beta proteins. RESULTS: NF-kappaB/Rel activity induced by tumor necrosis factor alpha, 12-O-tetradecanoylphorbol-13-acetate, or overexpression of NF-kappaB-inducing kinase, IKK-alpha, IKK-beta, or constitutively active IKK-alpha and IKK-beta mutants was inhibited dose dependently by sulfasalazine. Sulfasalazine inhibited tumor necrosis factor alpha-induced activation of endogenous IKK in Jurkat T cells and SW620 colon cells, as well as the catalytic activity of purified IKK alpha and IKK-beta in vitro. In contrast, the moieties of sulfasalazine, 5 aminosalicylic acid, and sulfapyridine or 4-aminosalicylic acid had no effect. Activation of extracellular signal-related kinase (ERK) 1 and 2, c-Jun-N-terminal kinase (JNK) 1, and p38 was unaffected by sulfasalazine. The decrease in substrate phosphorylation by IKK-alpha and -beta is associated with a decrease in autophosphorylation of IKKs and can be antagonized by excess adenosine triphosphate. CONCLUSIONS: These data identify sulfasalazine as a direct inhibitor of IKK-alpha and -beta by antagonizing adenosine triphosphate binding. The suppression of NF-kappaB activation by inhibition of the IKKs contributes to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine. PMID- 11054379 TI - Colorectal cancer screening by detection of altered human DNA in stool: feasibility of a multitarget assay panel. AB - BACKGROUND & AIMS: Assay of altered DNA exfoliated into stool represents an intriguing approach to screen for colorectal neoplasia, but multiple markers must be targeted because of genetic heterogeneity. We explored the feasibility of a stool assay panel of selected DNA alterations in discriminating subjects with colorectal neoplasia from those without. METHODS: Freezer-archived stools were analyzed in blinded fashion from 22 patients with colorectal cancer, 11 with adenomas > or =1 cm, and 28 with endoscopically normal colons. After isolation of human DNA from stool by sequence-specific hybrid capture, assay targets included point mutations at any of 15 sites on K-ras, p53, and APC genes; Bat-26, a microsatellite instability marker; and highly amplifiable DNA. RESULTS: Analyzable human DNA was recovered from all stools. Sensitivity was 91% (95% confidence interval, 71%-99%) for cancer and 82% (48%-98%) for adenomas > or =1 cm with a specificity of 93% (76%-99%). Excluding K-ras from the panel, sensitivities for cancer were unchanged but decreased slightly for adenomas to 73% (39%-94%), while specificity increased to 100% (88%-100%). CONCLUSIONS: Assay of altered DNA holds promise as a stool screening approach for colorectal neoplasia. Larger clinical investigations are indicated. PMID- 11054380 TI - JC virus DNA sequences are frequently present in the human upper and lower gastrointestinal tract. AB - BACKGROUND & AIMS: JC virus (JCV), a human polyomavirus, has been found in a limited number of normal human tissues and cancers. The oncogenic potential of this virus is mediated by a transforming protein, the T antigen (TAg). We have previously demonstrated the presence of JCV-TAg in colorectal cancers, in adjacent normal colonic mucosa from these patients, and in the human colon cancer cell line SW480. The mode of transmission of this virus is unclear, and we hypothesized that the gastrointestinal (GI) tract may be a reservoir for the virus. METHODS: DNA was extracted from 129 normal GI tissue samples collected from 33 patients. Topoisomerase I-assisted polymerase chain reaction (PCR) was used to detect the virus using exact and degenerate primers. Nested PCR and Southern blot analysis confirmed the identity of the PCR products. Single stranded conformation polymorphism (SSCP) analysis and sequencing were used to evaluate the presence of viral quasispecies. RESULTS: JCV sequences were found in 75.8% of patients (70.6% of upper GI and 81.2% of colonic samples); no significant differences in rates of infection were found by site. The use of degenerate primers combined with topoisomerase I treatment led to viral detection in 58.9% of samples, compared with 27.9% of samples using exact primers and topoisomerase I (P < 0.01). SSCP and sequencing analysis confirmed the amplification of viral quasispecies and the authenticity of TAg sequences. CONCLUSIONS: The results show that JCV DNA sequences are highly prevalent in the human upper and lower gastrointestinal tract of immunocompetent individuals. PMID- 11054381 TI - Mannose-binding lectin is a component of innate mucosal defense against Cryptosporidium parvum in AIDS. AB - BACKGROUND & AIMS: Nonimmune mechanisms of mucosal defense seem to be biologically important and might explain the observed variability in the course of enteric infection in immunodeficiency. Mannose-binding lectin (MBL) deficiency is associated with persistent diarrhea in children. We found that genetic determinants of MBL deficiency appear to predispose to cryptosporidiosis in patients with the acquired immunodeficiency syndrome (AIDS), and went on to study interactions of MBL and complement on Cryptosporidium parvum sporozoites. METHODS: This study involved cross-sectional study of MBL genotype and enteric infection in 72 Zambian AIDS patients with diarrhea, immunofluorescence analysis of MBL and C4 binding to C. parvum, and immunoblotting for MBL and complement in small intestinal fluid. RESULTS: Individuals homozygous for MBL structural gene mutations were at increased risk of cryptosporidiosis (odds ratio, 8.2; 95% confidence interval, 1. 5-42; P = 0.02). Lectin-mediated and concentration dependent binding of purified MBL was detected on sporozoites but not oocysts, and MBL activated C4 on sporozoites. MBL, C3, C4, and albumin were detected in small intestinal fluid in half the patients tested, suggesting transudation of serum components into the enteropathic gut. CONCLUSIONS: The increased risk of cryptosporidiosis in MBL deficiency appears to include patients with AIDS. It may operate through MBL-mediated complement activation on sporozoites. PMID- 11054382 TI - Group I secreted PLA2 in the maintenance of human lower esophageal sphincter tone. AB - BACKGROUND & AIMS: In cat spontaneous lower esophageal sphincter (LES), tone is maintained by the activity of group I secreted phospholipase A2 (sPLA2-I) that produces arachidonic acid. Arachidonic acid metabolites activate G proteins linked to phospholipases, producing second messengers and activation of a protein kinase C-dependent pathway to maintain tone. We examined the role of sPLA2-I in the maintenance of tone in human LES samples obtained from organ donors. METHODS: In vitro LES tone and sPLA2-I-induced contraction of enzymatically isolated LES smooth muscle cells were measured in the absence or presence of inhibitors. Cell permeabilization by saponin allowed use of G-protein antibodies. RESULTS: In vitro LES tone was reduced by inhibitors of sPLA2-I, by indomethacin, by the phosphatidylcholine-specific phospholipase C inhibitor D609, and by the protein kinase C inhibitor chelerythrine. sPLA2-I-induced contraction of isolated LES smooth muscle cells was reduced by indomethacin, pertussis toxin, Gi3 antibodies, D609, and by chelerythrine. CONCLUSIONS: Human LES tone is maintained by the activity of sPLA2-I that produces arachidonic acid and metabolites and activation of Gi3-linked receptors and of phosphatidylcholine-specific phospholipase C, resulting in production of diacylglycerol, activation of PKC, and maintenance of tone through a protein kinase C-dependent contractile pathway. PMID- 11054383 TI - IgA and IgM V(H) repertoires in human colon: evidence for clonally expanded B cells that are widely disseminated. AB - BACKGROUND & AIMS: The mucosal immune system defends the body from pathogens to which the mucosal surfaces are continually exposed. Because lamina propria B cells should reflect the antigenic experience of the gut, we investigated their immunoglobulin (Ig) repertoire and distribution. METHODS: The junctional diversity of the IgA and IgM heavy-chain transcripts in the colon and the peripheral blood of healthy adults was analyzed by CDR3 size spectratyping and nucleotide sequencing. RESULTS: The V(H)6 and V(H)7 repertoires of intestinal IgA and IgM cells were oligoclonal, whereas the CDR3 profiles of the larger V(H)1 V(H)5 families suggested a more diverse repertoire with dominant bands superimposed on a polyclonal background. However, sequence analysis revealed multiple repetitive and clonally related transcripts at distant colonic sites from all V(H) families. This suggests that, in addition to a polyclonal B-cell pool, subsets of B cells are clonally expanded and widely distributed along the colon. Occasionally, there was evidence for B cells with the same CDR3 specificity, which exhibited an isotype switch from IgM to IgA. Circulating IgA B cells expressed a restricted V(H) repertoire that was distinct from that in the colon. CONCLUSIONS: The human colon contains widely disseminated B cells that express clonally related IgA or IgM receptors. These results are best explained by an antigen-driven process whereby intestinal memory B cells continuously recirculate. PMID- 11054384 TI - 5-HT(3) and histamine H(1) receptors mediate afferent nerve sensitivity to intestinal anaphylaxis in rats. AB - BACKGROUND & AIMS: The mechanisms underlying brain stem activation during antigen challenge have not been resolved. Our aim was to characterize afferent nerve responses to intestinal anaphylaxis and determine the mediators involved in afferent activation. METHODS: Mesenteric afferent discharge was recorded electrophysiologically after intestinal anaphylaxis in anesthetized rats previously sensitized to chicken egg albumin (EA). RESULTS: Mesenteric afferent nerve discharge increased approximately 1 minute after luminal antigen but not bovine serum albumin (P < 0.001, EA vs. bovine serum albumin). Subsequent administration of antigen had no effect, but systemic EA evoked a marked increase in afferent discharge (P < 0. 05). Afferent responses were unrelated to intestinal motor activity, and the response to luminal antigen was attenuated by luminal anesthetic (1% lidocaine). The 5-HT(3)-receptor antagonist alosetron (30 microg. kg(-1)) and the histamine H(1)-receptor antagonist pyrilamine (5 mg. kg( 1)) markedly attenuated the response to luminal antigen; pretreatment with doxantrazole attenuated responses to both luminal and systemic antigen. CONCLUSIONS: 5-HT(3) and histamine, released from mast cells after intestinal anaphylaxis, stimulate mesenteric afferents via 5-HT(3) and histamine H(1) receptors. Information on intestinal immune status is rapidly relayed to the central nervous system and may play a role in neural reflexes and behavioral responses following activation of the immune system. PMID- 11054385 TI - A new model of chronic visceral hypersensitivity in adult rats induced by colon irritation during postnatal development. AB - BACKGROUND & AIMS: The irritable bowel syndrome (IBS) is a common disorder characterized by abdominal pain in the setting of altered perception of viscerosensory stimuli. This so-called visceral hyperalgesia occurs in the absence of detectable organic disease in the peripheral organs and may cause normal or physiologic contractions to be perceived as painful. Although the pathogenesis of IBS remains speculative and is probably multifactorial, a prevailing paradigm is that transient noxious events lead to long-lasting sensitization of the neural pain circuit, despite complete resolution of the initiating event. METHODS: Neonatal male Sprague-Dawley rats received either mechanical or chemical colonic irritation between postnatal days 8 and 21 and were tested when they became adults. The abdominal withdrawal reflex and the responses of viscerosensitive neurons were recorded during colon distention. RESULTS: Colon irritation in neonates, but not in adults, results in chronic visceral hypersensitivity, with characteristics of allodynia and hyperalgesia, associated with central neuronal sensitization in the absence of identifiable peripheral pathology. CONCLUSIONS: These results concur largely with observations in patients with IBS, providing a new animal model to study IBS and validating a neurogenic component of functional abdominal pain that encourages novel approaches to health care and research. PMID- 11054386 TI - Recombinant soluble transforming growth factor beta type II receptor ameliorates radiation enteropathy in mice. AB - BACKGROUND & AIMS: Transforming growth factor (TGF)-beta has been implicated in many fibrotic conditions. However, its mechanistic role in radiation toxicity is equivocal despite compelling correlative evidence. This study assessed whether in vivo administration of a soluble TGF-beta type II receptor (TbetaR-II) protein ameliorates intestinal radiation injury (radiation enteropathy). METHODS: A recombinant fusion protein, consisting of the extracellular portion of mouse TbetaR-II and the Fc portion of mouse immunoglobulin (Ig) G, was produced. A 5-cm segment of mouse ileum was exposed to 19 Gy x-radiation. TbetaR-II:Fc fusion protein (1 mg/kg every other day) or mouse IgG was administered from 2 days before to 6 weeks after irradiation. Radiation injury was assessed at 6 weeks using quantitative histology, morphometry, and immunohistochemistry. Collagen was measured colorimetrically, and TGF-beta1 messenger RNA was assessed with fluorogenic probe reverse-transcription polymerase chain reaction. RESULTS: Compared with IgG controls, TbetaR-II:Fc-treated mice exhibited less structural injury, preservation of mucosal surface area, and less intestinal wall fibrosis. Intestinal TGF-beta1 messenger RNA increased in TbetaR-II:Fc-treated mice, whereas TGF-beta immunoreactivity decreased. TbetaR-II:Fc treatment increased crypt cell proliferation but otherwise did not affect unirradiated intestine. CONCLUSIONS: Long-term modulation of TGF-beta with a TbetaR-II:Fc fusion protein is feasible and ameliorates radiation enteropathy. These data confirm the putative role of TGF-beta in intestinal radiation fibrosis. PMID- 11054387 TI - Acid transport through channels in the mucous layer of rat stomach. AB - BACKGROUND & AIMS: We have reported previously that secreted acid moves through the mucous layer in restricted areas above the gastric crypts. The aim of this study was to visualize and study the dynamics of this event. METHODS: Anesthetized rats prepared for intravital microscopy of the gastric mucosa were divided in the following groups with respect to acid secretion: spontaneous; stimulated (pentagastrin, 40 microg. kg(-1). h(-1)); transiently inhibited (omeprazole, 400 micromol. kg(-1) for 7 days); and totally inhibited (omeprazole, 3 x 200 micromol. kg(-1) for 7 days). The mucus was stained with Congo red (blue, pH < 3; red, pH > 5.2), and photographs were taken through a stereomicroscope. RESULTS: During acid secretion, blue-colored crypt openings with attached thread like (5-7 microm wide) structures (designated channels) were seen passing from the crypt openings through the mucus to the lumen. Red-colored channels and crypt openings were observed when acid secretion was transiently inhibited. Red-colored crypt openings but no channels were found after total inhibition of acid secretion for a week. CONCLUSIONS: The results suggest that secreted acid is transported through channels within the mucus. These channels are probably created by the high intraglandular pressure pushing acid and glandular mucus into the gel. PMID- 11054388 TI - Role of the amino-terminal domain of simian virus 40 early region in inducing tumors in secretin-expressing cells in transgenic mice. AB - BACKGROUND & AIMS: The early region of simian virus 40 (SV40) encodes 2 transforming proteins, large T (Tag) and small t antigen, that produce neuroendocrine tumors in the intestine and the pancreas when expressed in secretin cells of transgenic mice. METHODS: Two SV40 early-region transgenes containing a deletion that eliminated expression of the small t antigen were expressed in transgenic mice under control of the secretin gene. The 2 lines of mice, one expressing the native large T antigen and the other T antigen with a mutation in its N-terminal J domain, were examined to determine which biological activities of the SV40 early region were required for tumorigenesis. RESULTS: Most animals expressing wild-type large T antigen developed pancreatic insulinomas and lymphomas and died between 3 and 6 months of age. However, small intestinal neoplasms were extremely rare in the absence of small t antigen expression. Transgenic lines expressing the J domain mutant failed to develop tumors. CONCLUSIONS: Transformation of secretin-producing enteroendocrine cells by SV40 requires functional cooperation between intact large T and small t oncoproteins. In contrast, large T antigen alone is sufficient to induce tumors in the endocrine pancreas and thymus. PMID- 11054389 TI - Failure of budesonide in a pilot study of treatment-dependent autoimmune hepatitis. AB - BACKGROUND & AIMS: Budesonide has a high hepatic first-pass clearance and metabolites virtually devoid of glucocorticoid activity. Our goals were to assess budesonide in patients with treatment-dependent autoimmune hepatitis and to determine if efficacy and safety justified a controlled trial. METHODS: Ten patients who were dependent on continuous treatment to prevent exacerbation of their disease were treated with budesonide, 3 mg thrice daily. RESULTS: Laboratory indices did not improve significantly during 5 +/- 1 months of therapy (range, 2-12 months). Three patients entered clinical and biochemical remission; 2 of them achieved histologic remission. Seven patients either deteriorated during therapy or became drug intolerant. Withdrawal symptoms complicated conversion from prednisone to budesonide treatment, and every patient developed at least 1 side effect. Lumbar spine density decreased in 2 patients, and femur density decreased in 2 patients, including 1 with lumbar spine changes. However, mean bone densities actually increased slightly in the entire group. CONCLUSIONS: Budesonide therapy was associated with a low frequency of remission and high occurrence of treatment failure and side effects in treatment-dependent autoimmune hepatitis. Findings did not support the need for a controlled treatment trial in this select population. PMID- 11054390 TI - The impact of interferon plus ribavirin on response to therapy in black patients with chronic hepatitis C. The International Hepatitis Interventional Therapy Group. AB - BACKGROUND & AIMS: Black patients with chronic hepatitis C have lower response rates than white patients to interferon monotherapy. The factors responsible for these differences are unknown, as is the impact of combination antiviral therapy on responsiveness among ethnic groups. We evaluated the impact of race on response to therapy in these patients. METHODS: A total of 1744 patients with chronic hepatitis C were randomized in 2 recent clinical trials to receive 24 or 48 weeks of interferon monotherapy or interferon-ribavirin combination therapy. RESULTS: Sustained virologic responses occurred in 27% of 1600 whites, 11% of 53 blacks (P = 0.01 vs. white), 44% of 32 Asians, and 16% of 27 Hispanics. No black patient had a sustained virologic response to interferon monotherapy, but 20% and 23% had sustained responses to 24 and 48 weeks, respectively, of combination therapy. Among black patients, 96% had hepatitis C genotype 1 compared with 65% of white subjects (P < 0.0001). Sustained response rates were similar for black and white patients with genotype 1 infection (23% vs. 22%, respectively). Compared with whites, black patients were older, weighed more, and had higher median Histologic Activity Index scores but did not differ in sex, baseline alanine aminotransferase or hepatitis C virus (HCV)-RNA levels, degree of fibrosis or percentage with cirrhosis, or other demographic variables. White subjects had a significantly greater reduction in HCV-RNA levels than blacks at weeks 4, 12, 24, and 48 of therapy, but only for black patients treated with interferon monotherapy. The decreased reduction of HCV-RNA reduction among blacks was eliminated by combination therapy. CONCLUSIONS: These observations suggest that the impaired responsiveness of black patients to interferon monotherapy can be overcome partially by combination interferon-ribavirin therapy. PMID- 11054391 TI - Risk factors for diabetes mellitus in chronic pancreatitis. AB - BACKGROUND & AIMS: The influence of disease progression and pancreatic surgery on the appearance of diabetes mellitus in patients with chronic pancreatitis is unknown. METHODS: A prospective cohort study of 500 consecutive patients with chronic pancreatitis (alcoholics, 85%) followed up over a mean period of 7.0 +/- 6.8 years in a medical-surgical institution between 1973 and 1996 was performed. Multivariate analysis of risk factors for diabetes mellitus was performed after exclusion of 47 patients. Patients who underwent elective pancreatic surgery (n = 231, 51%) were compared with patients who never underwent surgery (n = 222, 49%). RESULTS: The cumulative rate of diabetes mellitus was 83% +/- 4% 25 years after the clinical onset of chronic pancreatitis (insulin requirement, 54% +/- 6%). The prevalence of diabetes mellitus did not increase in the surgical group overall but was higher 5 years after distal pancreatectomy (57% +/- 8%) than after pancreaticoduodenectomy (36% +/- 18%), pancreatic drainage (36% +/- 13%), or cystic, biliary, or digestive drainage (24% +/- 7%) (P = 0. 005), without difference in the latter ones. Pancreatic drainage did not prevent the onset of diabetes mellitus. Distal pancreatectomy (risk ratio, 2.4; 95% confidence interval [CI], 1.6-3.8; P < 0.0001) and early onset of pancreatic calcifications (risk ratio, 3.2; CI, 2. 2-4.7; P < 0.0001) were the only independent risk factors for diabetes mellitus. CONCLUSIONS: The risk of diabetes mellitus is not influenced by elective pancreatic surgical procedures other than distal pancreatectomy in patients with chronic pancreatitis. This risk seems to be largely caused by progression of the disease because it increased by more than 3 fold after the onset of pancreatic calcifications. PMID- 11054392 TI - Light ethanol consumption enhances liver regeneration after partial hepatectomy in rats. AB - BACKGROUND & AIMS: The effects of "social drinking" on the liver have yet to be fully documented. The aim of this study was to document the effects of daily light, moderate, and heavy ethanol exposure on hepatic regenerative activity in the rat. METHODS: Adult male Sprague-Dawley rats underwent daily gavages with 1.0 (light), 2. 0 (moderate), or 4.0 (heavy) g/kg of ethanol or tap water (controls) for 30 days before 70% partial hepatectomy (PHx). Hepatic regenerative activity was then documented on days 1, 3, 5, and 7 after PHx. RESULTS: Compared with controls, restitution of liver mass, [(3)H]thymidine incorporation, and proliferating cell nuclear antigen expression were decreased in the heavy (-10%, 60%, and -36%, respectively), unchanged in the moderate (-4%, -8%, and -16%, respectively), and increased in the light (+6%, +38%, and +29%, respectively) ethanol groups. Messenger RNA differential display of resected livers at PHx identified a band present only in the light ethanol group that encodes a unique 47-kilodalton protein with growth-promoting features designated light ethanol induced stimulatory protein. CONCLUSIONS: The results indicate that light ethanol consumption enhances hepatic regenerative activity after PHx in rats. Further studies are required to determine the mechanism involved and whether social drinking has beneficial or adverse effects on the natural history of acute or chronic liver disease in humans. PMID- 11054393 TI - Increased gastrointestinal ethanol production in obese mice: implications for fatty liver disease pathogenesis. AB - BACKGROUND & AIMS: Similarities in the hepatic responses to obesity and ethanol exposure suggest that these conditions evoke common pathogenic mechanisms. Thus, it is possible that ethanol exposure is increased in obesity. Given that intestinal bacteria can produce ethanol, the aim of this study was to determine if the intestinal production of ethanol is increased in obesity. METHODS: Breath was collected from genetically obese, ob/ob male C57BL/6 mice and lean male littermates at different ages (14, 20, and 24 weeks) and times of the day (9 AM, 3 PM, and 9 PM). Obese mice (24 weeks old) were then treated with neomycin (1 mg/mL) for 5 days, and sampling was repeated. RESULTS: Breath collected in the morning from 24-week-old obese mice had a higher ethanol content than breath from their lean littermates (271 vs. 78 pmol/mL CO(2); P < 0.0001). Subsequent studies in 14- and 20-week-old mice showed that exhaled ethanol increased with age in obese (from 26 to 107 pmol/mL CO(2); P < 0. 002) but not lean (29 and 12 pmol/mL CO(2)) mice and was greater in older obese mice than in older lean mice (P < 0.0006). Obese mice showed a diurnal increase in breath ethanol in the morning that decreased through the afternoon and evening (107 to 33 to 13 pmol/mL CO(2)). Neomycin treatment decreased morning breath ethanol levels by 50% (from 220 to 110 pmol/mL CO(2); P < 0.0003). CONCLUSIONS: Even in the absence of ethanol ingestion, ethanol can be detected in exhaled breath. In obesity, an age-related increase in breath ethanol content reflects increased production of ethanol by the intestinal microflora. Hence, intestinal production of ethanol may contribute to the genesis of obesity-related fatty liver. PMID- 11054394 TI - A replication-deficient rSV40 mediates liver-directed gene transfer and a long term amelioration of jaundice in gunn rats. AB - BACKGROUND & AIMS: In the quest for a recombinant viral vector for liver-directed gene therapy that would permit both prolonged and efficient transgene expression in quiescent hepatocytes in vivo and repeated administration, we evaluated a recombinant simian virus 40 (rSV40). METHODS: The rSV40 was generated through replacement of the DNA encoding for the T antigens (Tag) by the coding region of human bilirubin-uridine 5'-diphosphate-glucuronosyl-transferase (BUGT) complementary DNA (SV-hBUGT). Helper-free rSV40 units were generated at infectious titers of 5 x 10(9) to 1 x 10(10) infectious units (IU)/mL in a Tag producing packaging cell line (COS-7 cells). RESULTS: After 1, 3, or 7 daily infusions of 3 x 10(9) IU of SV-hBUGT through an indwelling portal vein catheter in bilirubin-UGT-deficient jaundiced Gunn rats, mean serum bilirubin concentrations decreased by 40%, 60% and 70%, respectively, in 3 weeks and remained at those levels throughout the duration of the study (40 days). Results of liver biopsies from SV-hBUGT-treated Gunn rats, but not from controls, were positive for human BUGT DNA, messenger RNA, and protein. Bilirubin-UGT activity in liver homogenates was 8%-12% of normal, and bilirubin glucuronides were excreted in bile. Immunostaining showed that >50%-60% of hepatocytes stably expressed the transgene. Portal vein infusion of an rSV40 expressing hepatitis B surface antigen (HBsAg) in a naive Gunn rat and a Gunn rat that had received 7 injections of SV-BUGT resulted in approximately equal levels of hepatic expression of HBsAg, indicating that multiple inoculations of SV-BUGT did not elicit neutralizing antibodies. Plasma alanine aminotransferase levels and liver histology remained normal despite repeated injections of rSV40. CONCLUSIONS: rSV40 vectors may represent a significant advance toward gene therapy for metabolic diseases. PMID- 11054395 TI - De novo expression of vascular endothelial growth factor in human pancreatic cancer: evidence for an autocrine mitogenic loop. AB - BACKGROUND & AIMS: The role of vascular endothelial growth factor (VEGF) and its receptors in tumor angiogenesis has been well established. We analyzed the expression pattern and biologic significance of VEGF and its receptors in human pancreatic cancer. METHODS: VEGF, KDR/flk-1, and flt-1 expression were examined by immunohistochemistry, in situ hybridization, reverse-transcription polymerase chain reaction, enzyme-linked immunosorbent assay, and receptor phosphorylation. VEGF-stimulated mitogenesis was investigated by mitogen-activated protein kinase (MAPK) phosphorylation, transactivation of a c-fos promoter reporter construct, DNA synthesis assays, and stable transfection of a dominant-negative flk-1 complementary DNA (cDNA) construct. RESULTS: Compared with normal pancreas and chronic pancreatitis, VEGF and its receptors were overexpressed in pancreatic cancer. KDR and flt-1 were detected not only in endothelial cells but also in tumor cells. VEGF expression was observed in all human pancreatic tumor cell lines examined, and the KDR/flk-1 and flt-1 receptor was detected in 2 cell lines. VEGF treatment results in phosphorylation of MAPKs, transactivation of a c fos promoter construct, and growth stimulation in KDR/flk-1-expressing cell lines, which could be blocked by VEGF antagonists. Furthermore, stable transfection of a dominant-negative flk-1 cDNA significantly inhibited tumor cell growth. CONCLUSIONS: These results not only support the important role of the VEGF/VEGF receptor system in pancreatic tumor biology but also suggest the existence of an autocrine/paracrine mitogenic loop for pancreatic cancer cells. PMID- 11054396 TI - Nerve growth factor expression is up-regulated in the rat model of L-arginine induced acute pancreatitis. AB - BACKGROUND & AIMS: In somatic pain models, increases in nerve growth factor (NGF) are linked to the development of pain and hyperalgesia. The aim of this study was to examine a rat model of acute necrotizing pancreatitis for changes in NGF expression. METHODS: NGF protein and messenger RNA (mRNA) levels in the pancreas were correlated with histopathologic changes during the course of acute necrotizing pancreatitis in rats induced by the intraperitoneal injection of L arginine. Immunohistochemistry for NGF localization was performed on the pancreatic tissue. RESULTS: Two phases of NGF production were observed in the inflamed pancreas: an early release from pancreatic islets at 2 and 6 hours and a later increase in mRNA (18-fold at maximum) at 3 days and in protein levels (7 fold at maximum) at 5 days coinciding with maximum parenchymal necrosis. The intense NGF-like immunoreactivity was observed predominantly in the ductal cells in pancreas from rats with pancreatitis at 5 days. CONCLUSIONS: The development of acute necrotizing pancreatitis in this model leads to a significant increase in NGF production and appears to shift the major cellular sites of NGF production from the islets to the ductal cells. It is conceivable that NGF production in the inflamed pancreas is responsible for plastic changes in the sensory neurons that mediate peripheral sensitization and contribute to the generation of pain. PMID- 11054397 TI - Successful orthotopic liver transplantation for lamivudine-associated YMDD mutant hepatitis B virus. AB - YMDD hepatitis B virus (HBV) mutants emerge in more than one third of patients treated with lamivudine. The development of escape mutants potentially limits the efficacy of this new therapy for chronic HBV infection. Transplant centers may be hesitant to perform orthotopic liver transplantation (OLT) in patients with progressive liver disease who develop the YMDD mutant because of concerns of graft reinfection. We describe a 56-year-old man with chronic liver disease caused by HBV who successfully underwent OLT after the emergence of a YMDD mutant and progressive liver disease. Perioperatively, he received high-dose intravenous hepatitis B immune globulin (HBIG) treatment with continued lamivudine treatment. Thirty-two months after OLT, there was no evidence of HBV reinfection with the use of lamivudine and high-dose HBIG. Our experience suggests that the emergence of a YMDD HBV mutant is not a contraindication to OLT. The combination of lamivudine and high-dose HBIG can protect against reinfection of the hepatic allograft with the YMDD HBV escape mutant. PMID- 11054398 TI - Hepatitis C in African Americans: summary of a workshop. PMID- 11054399 TI - Unlocking the secrets of the porcelain vase. PMID- 11054400 TI - Moderate alcohol drinking: effects on the heart and liver. PMID- 11054401 TI - Colonoscopy versus double-contrast barium enema. PMID- 11054402 TI - Surveying the masses. PMID- 11054403 TI - Hepatitis C for half a century. PMID- 11054405 TI - Hemochromatosis: genetics, pathopysiology, diagnosis and treatment PMID- 11054404 TI - Peritoneal dialysis: an under-appreciated cause of acute pancreatitis. PMID- 11054406 TI - Regulation of plasminogen activator inhibitor-1 expression by transforming growth factor-beta -induced physical and functional interactions between smads and Sp1. AB - Members of the transforming growth factor-beta (TGF-beta) superfamily mediate a broad range of biological activities by regulating the expression of target genes. Smad proteins play a critical role in this process by binding directly to the promoter elements and/or associating with other transcription factors. TGF beta1 up-regulates several genes transcriptionally through Sp1 binding sites; however, the mechanism of TGF-beta induction of gene expression through Sp1 sites is largely unknown. Here we report the identification of a novel 38-base pair TGF beta-responsive element in the human plasminogen activator inhibitor-1 (PAI-1) promoter, which contains two Sp1 binding sites, and is required for TGF-beta induced Smad-dependent transcriptional activation. Three canonical Sp1 binding sites also support strong transcriptional activation by TGF-beta and Smads from a minimal heterologous promoter. TGF-beta induction of PAI-1 and p21 is blocked by the Sp1 inhibitor mithramycin, implicating Sp1 in the in vivo regulation of these genes by TGF-beta. We show that the association between endogenous Sp1 and Smad3 is induced by TGF-beta in several cell lines; however, Smad4 shows constitutive interaction with Sp1. These data provide novel insights into the mechanism by which TGF-beta up-regulates several gene expression by activating Sp1-dependent transcription through the induction of Smad/Sp1 complex formation. PMID- 11054407 TI - Transcriptional analysis of chromatin assembled with purified ACF and dNAP1 reveals that acetyl-CoA is required for preinitiation complex assembly. AB - To investigate the role of chromatin structure in the regulation of transcription by RNA polymerase II, we developed a chromatin transcription system in which periodic nucleosome arrays are assembled with purified recombinant ATP-utilizing chromatin assembly and remodeling factor (ACF), purified recombinant nucleosome assembly protein 1 (dNAP1), purified native core histones, plasmid DNA, and ATP. With this chromatin, we observed robust activation of transcription with three different transcription factor sets (nuclear factor kappaB p65 + Sp1, estrogen receptor, and Gal4-VP16) added either before or after chromatin assembly. In fact, the efficiency of activated transcription from the ACF + dNAP1-assembled chromatin was observed to be comparable with that from naked DNA templates or chromatin assembled with a crude Drosophila extract (S190). With ACF + dNAP1 assembled chromatin, we found that transcriptional activation is dependent upon acetyl-CoA. This effect was not seen with naked DNA templates or with crude S190 assembled chromatin. We further determined that acetyl-CoA is required at the time of preinitiation complex assembly but not during assembly of the chromatin template. These findings suggest that there is at least one key acetylation event that is needed to assemble a functional transcription preinitiation complex with a chromatin template. PMID- 11054408 TI - Thrombopoietin induces phosphoinositol 3-kinase activation through SHP2, Gab, and insulin receptor substrate proteins in BAF3 cells and primary murine megakaryocytes. AB - Thrombopoietin (TPO) is a recently characterized member of the hematopoietic growth factor family that serves as the primary regulator of megakaryocyte (MK) and platelet production. The hormone acts by binding to the Mpl receptor, the product of the cellular proto-oncogene c-mpl. Although many downstream signaling targets of TPO have been identified in cell lines, primary MKs, and platelets, the molecular mechanism(s) by which many of these molecules are activated remains uncertain. In this report we demonstrate that the TPO-induced activation of phosphoinositol 3-kinase (PI3K), a signaling intermediate vital for cellular survival and proliferation, occurs through its association with inducible signaling complexes in both BaF3 cells engineered to express Mpl (BaF3/Mpl) and in primary murine MKs. Although a direct association between PI3K and Mpl could not be demonstrated, we found that several proteins, including SHP2, Gab2, and IRS2, undergo phosphorylation and association in BaF3/Mpl cells in response to TPO stimulation, complexes that recruit and enhance the enzymatic activity of PI3K. To verify the physiological relevance of the complex, SHP2-Gab2 association was disrupted by overexpressing a dominant negative SHP2 construct. TPO-induced Akt phosphorylation was significantly decreased in transfected cells suggesting an important role of SHP2 in the complex to enhance PI3K activity. In primary murine MKs, TPO also induced phosphorylation of SHP2, its association with p85 and enhanced PI3K activity, but in contrast to the results in cell lines, neither Gab2 nor IRS2 are phosphorylated in MKs. Instead, a 100-kDa tyrosine phosphorylated protein (pp100) co-immunoprecipitated with the regulatory subunit of PI3K. These findings support a model where PI3K activity is dependent on its recruitment into TPO-induced multiphosphoprotein complexes, implicate the existence of a scaffolding protein in primary MKs distinct from the known Gab and IRS proteins, and suggest that, in contrast to erythroid progenitor cells that employ Gab1 in PI3K signaling complexes, utilization of an alternate member of the Gab/IRS family could be responsible for specificity in TPO signaling. PMID- 11054409 TI - Hypoxia and nitric oxide induce a rapid, reversible cell cycle arrest of the Drosophila syncytial divisions. AB - Cells can respond to reductions in oxygen (hypoxia) by metabolic adaptations, quiescence or cell death. The nuclear division cycles of syncytial stage Drosophila melanogaster embryos reversibly arrest upon hypoxia. We examined this rapid arrest in real time using a fusion of green fluorescent protein and histone 2A. In addition to an interphase arrest, mitosis was specifically blocked in metaphase, much like a checkpoint arrest. Nitric oxide, recently proposed as a hypoxia signal in Drosophila, induced a reversible arrest of the nuclear divisions comparable with that induced by hypoxia. Syncytial stage embryos die during prolonged hypoxia, whereas post-gastrulation embryos (cellularized) survive. We examined ATP levels and morphology of syncytial and cellularized embryos arrested by hypoxia, nitric oxide, or cyanide. Upon oxygen deprivation, the ATP levels declined only slightly in cellularized embryos and more substantially in syncytial embryos. Reversal of hypoxia restored ATP levels and relieved the cell cycle and developmental arrests. However, morphological abnormalities suggested that syncytial embryos suffered irreversible disruption of developmental programs. Our results suggest that nitric oxide plays a role in the response of the syncytial embryo to hypoxia but that it is not the sole mediator of these responses. PMID- 11054410 TI - Utilization of sialic acid as a coreceptor enhances reovirus attachment by multistep adhesion strengthening. AB - Many serotype 3 reoviruses bind to two different host cell molecules, sialic acid and an unidentified protein, using discrete receptor-binding domains in viral attachment protein, final sigma1. To determine mechanisms by which these receptor binding events cooperate to mediate cell attachment, we generated isogenic reovirus strains that differ in the capacity to bind sialic acid. Strain SA+, but not SA-, bound specifically to sialic acid on a biosensor chip with nanomolar avidity. SA+ displayed 5-fold higher avidity for HeLa cells when compared with SA , although both strains recognized the same proteinaceous receptor. Increased avidity of SA+ binding was mediated by increased k(on). Neuraminidase treatment to remove cell-surface sialic acid decreased the k(on) of SA+ to that of SA-. Increased k(on) of SA+ enhanced an infectious attachment process, since SA+ was 50-100-fold more efficient than SA- at infecting HeLa cells in a kinetic fluorescent focus assay. Sialic acid binding was operant early during SA+ attachment, since the capacity of soluble sialyllactose to inhibit infection decreased rapidly during the first 20 min of adsorption. These results indicate that reovirus binding to sialic acid enhances virus infection through adhesion of virus to the cell surface where access to a proteinaceous receptor is thermodynamically favored. PMID- 11054411 TI - PRH represses transcription in hematopoietic cells by at least two independent mechanisms. AB - PRH (proline-rich homeodomain protein) is strongly expressed in the hematopoietic compartment. Here we show that PRH is a repressor of transcription in hematopoietic cells. A fragment of PRH that includes the homeodomain can bind to TATA box sequences in vitro and can also bind to the TATA box-binding protein. PRH represses transcription from TATA box-containing promoters in intact cells but does not repress transcription from a promoter lacking a TATA box. A mutation in the PRH homeodomain that blocks binding to DNA but that has little or no effect on binding to the TATA box-binding protein significantly reduces the ability of the protein to repress transcription and provides the first clear demonstration that a homeodomain can bring about transcriptional repression in vivo by binding to a TATA box. However, we also show that mutation of the PRH homeodomain does not block the ability of PRH to repress transcription when this protein is tethered upstream of the TATA box via a heterologous DNA-binding domain. PRH also contains an N-terminal proline-rich repression domain that is separate from the homeodomain. Deletion mapping suggests that this repression domain contains at least two regions that both independently contribute to transcriptional repression. PMID- 11054412 TI - Selectively suppressed Ca2+-induced Ca2+ release activity of alpha-ryanodine receptor (alpha-RyR) in frog skeletal muscle sarcoplasmic reticulum: potential distinct modes in Ca2+ release between alpha- and beta-RyR. AB - We reported earlier that the two ryanodine receptor (RyR) isoforms (alpha- and beta-RyR) purified from frog skeletal muscle were equipotent in the Ca(2+) induced Ca(2+) release (CICR) activity (Murayama, T., Kurebayashi, N., and Ogawa, Y. (2000) Biophys. J. 78, 1810-1824). Whether this is also the case with the native Ca(2+) release channel in the sarcoplasmic reticulum (SR), however, remains to be determined. Taking advantage of the facts that [(3)H]ryanodine binds only to the open form of the channels and that it is practically irreversible at 4 degrees C, we devised a method to separate the total binding to contributions of alpha- and beta-RyR, using immunoprecipitation with an alpha-RyR specific monoclonal antibody. Surprisingly, the binding of alpha-RyR was strongly suppressed to as low as approximately 4% that of beta-RyR in the SR vesicles. The two isoforms, however, showed no difference in sensitivity to Ca(2+), adenine nucleotides, or caffeine. This reduced binding of alpha-RyR was ascribed to the low affinity for [(3)H]ryanodine, with no change in the maximal binding sites. Solubilization of SR with 3-[(3-cholamidopropyl) dimethylammonio]-1 propanesulfonic acid partly remedied this nonequivalence, whereas 1 m NaCl was ineffective. 12-kDa FK506-binding protein (FKBP12), however, could not be responsible for it, because FK506 treatment did not eliminate the suppression, in contrast to marked removal of 12-kDa FK506-binding protein from alpha-RyR. These results suggest that alpha-RyR in the SR may serve Ca(2+) release in a mode other than CICR, being selectively suppressed in CICR. PMID- 11054413 TI - Bcl-G, a novel pro-apoptotic member of the Bcl-2 family. AB - A new member of the Bcl-2 family was identified, Bcl-G. The human BCL-G gene consists of 6 exons, resides on chromosome 12p12, and encodes two proteins through alternative mRNA splicing, Bcl-G(L) (long) and Bcl-G(S) (short) consisting of 327 and 252 amino acids in length, respectively. Bcl-G(L) and Bcl G(S) have identical sequences for the first 226 amino acids but diverge thereafter. Among the Bcl-2 homology (BH) domains previously recognized in Bcl-2 family proteins, the BH3 domain is found in both Bcl-G(L) and Bcl-G(S), but only the longer Bcl-G(L) protein possesses a BH2 domain. Bcl-G(L) mRNA is expressed widely in adult human tissues, whereas Bcl-G(S) mRNA was found only in testis. Overexpression of Bcl-G(L) or Bcl-G(S) in cells induced apoptosis although Bcl G(S) was far more potent than Bcl-G(L). Apoptosis induction by Bcl-G(S) depended on the BH3 domain and was suppressed by coexpression of anti-apoptotic Bcl-X(L) protein. Bcl-X(L) also coimmunoprecipitated with Bcl-G(S) but not with mutants of Bcl-G(S) in which the BH3 domain was deleted or mutated or with Bcl-G(L). Bcl G(S) was predominantly localized to cytosolic organelles, whereas Bcl-G(L) was diffusely distributed throughout the cytosol. A mutant of Bcl-G(L) in which the BH2 domain was deleted displayed increased apoptotic activity and coimmunoprecipitated with Bcl-X(L), suggesting that the BH2 domain autorepresses Bcl-G(L). PMID- 11054414 TI - Lipid oxidation enhances the function of activated protein C. AB - Although lipid oxidation products are usually associated with tissue injury, it is now recognized that they can also contribute to cell activation and elicit anti-inflammatory lipid mediators. In this study, we report that membrane phospholipid oxidation can modulate the hemostatic balance. Oxidation of natural phospholipids results in an increased ability of the membrane surface to support the function of the natural anticoagulant, activated protein C (APC), without significantly altering the ability to support thrombin generation. Lipid oxidation also potentiated the ability of protein S to enhance APC-mediated factor Va inactivation. Phosphatidylethanolamine, phosphatidylserine, and polyunsaturation of the fatty acids were all required for the oxidation-dependent enhancement of APC function. A subgroup of thrombotic patients with anti phospholipid antibodies specifically blocked the oxidation-dependent enhancement of APC function. Since leukocytes are recruited and activated at the thrombus or sites of vessel injury, our findings suggest that after the initial thrombus formation, lipid oxidation can remodel the membrane surface resulting in increased anticoagulant function, thereby reducing the thrombogenicity of the thrombus or injured vessel surface. Anti-phospholipid antibodies that block this process would therefore be expected to contribute to thrombus growth and disease. PMID- 11054415 TI - Long terminal repeats are used as alternative promoters for the endothelin B receptor and apolipoprotein C-I genes in humans. AB - To examine the potential regulatory involvement of retroelements in the human genome, we screened the transcribed sequences of GenBank and expressed sequence tag data bases with long terminal repeat (LTR) elements derived from different human endogenous retroviruses. These screenings detected human transcripts containing LTRs belonging to the human endogenous retrovirus-E family fused to the apolipoprotein CI (apoC-I) and the endothelin B receptor (EBR) genes. However, both genes are known to have non-LTR (native) promoters. Initial reverse transcription-polymerase chain reaction experiments confirmed and authenticated the presence of transcripts from both the native and LTR promoters. Using a 5' rapid amplification of cDNA ends protocol, we showed that the alternative transcripts of apoC-I and EBR are initiated and promoted by the LTRs. The LTR apoC-I fusion and native apoC-I transcripts are present in many of the tissues tested. As expected, we found apoC-I preferentially expressed in liver, where about 15% of the transcripts are derived from the LTR promoter. Transient transfections suggest that the expression is not dependent on the LTR itself, but the presence of the LTR increases activity of the apoC-I promoter from both humans and baboons. The native EBR-driven transcripts were also detected in many tissues, whereas the LTR-driven transcripts appear limited to placenta. In contrast to the LTR of apoC-I, the EBR LTR promotes a significant proportion of the total EBR transcripts, and transient transfection results indicate that the LTR acts as a strong promoter and enhancer in a placental cell line. This investigation reports two examples where LTR sequences contribute to increased transcription of human genes and illustrates the impact of mobile elements on gene and genome evolution. PMID- 11054416 TI - Interorganellar communication. Altered nuclear gene expression profiles in a yeast mitochondrial dna mutant. AB - Communication between mitochondria and the nucleus is important for a variety of cellular processes such as carbohydrate and nitrogen metabolism, mating and sporulation, and cell growth and morphogenesis. It has long been known that the functional state of mitochondria can influence nuclear gene expression. For example, in yeast cells lacking the mitochondrial genome, the expression of several nuclear genes, such as CIT2 (citrate synthase), MRP13 (mitochondrial ribosomal protein), and DLD3 (d-lactate dehydrogenase) has been reported to be altered. Here we show by microarray analysis of the genome-wide transcription profile of Saccharomyces cerevisiae that yeast petite mutants lacking mitochondrial DNA induce genes coding for mitochondrial proteins, enzymes of the glycolytic pathway and of the citric acid cycle, cell wall components, membrane transporters, and genes normally induced by nutrient deprivation and a variety of stresses. Consistent with the observed induction of genes related to cell stress and those encoding membrane transporters, yeast petite cells showed increased resistance to severe heat shock and exhibited a pleiotropic drug resistance phenotype. The observed changes in nuclear gene expression in cells lacking mitochondrial DNA may have implications for the role of mitochondria in processes such as carcinogenesis and aging. PMID- 11054417 TI - Newly synthesized human delta opioid receptors retained in the endoplasmic reticulum are retrotranslocated to the cytosol, deglycosylated, ubiquitinated, and degraded by the proteasome. AB - We have previously shown that only a fraction of the newly synthesized human delta opioid receptors is able to leave the endoplasmic reticulum (ER) and reach the cell surface (Petaja-Repo, U. E, Hogue, M., Laperriere, A., Walker, P., and Bouvier, M. (2000) J. Biol. Chem. 275, 13727-13736). In the present study, we investigated the fate of those receptors that are retained intracellularly. Pulse chase experiments revealed that the disappearance of the receptor precursor form (M(r) 45,000) and of two smaller species (M(r) 42,000 and 39,000) is inhibited by the proteasome blocker, lactacystin. The treatment also promoted accumulation of the mature receptor form (M(r) 55,000), indicating that the ER quality control actively routes a significant proportion of rescuable receptors for proteasome degradation. In addition, degradation intermediates that included full-length deglycosylated (M(r) 39,000) and ubiquitinated forms of the receptor were found to accumulate in the cytosol upon inhibition of proteasome function. Finally, coimmunoprecipitation experiments with the beta-subunit of the Sec61 translocon complex revealed that the receptor precursor and its deglycosylated degradation intermediates interact with the translocon. Taken together, these results support a model in which misfolded or incompletely folded receptors are transported to the cytoplasmic side of the ER membrane via the Sec61 translocon, deglycosylated and conjugated with ubiquitin prior to degradation by the cytoplasmic 26 S proteasomes. PMID- 11054418 TI - Munc18c regulates insulin-stimulated glut4 translocation to the transverse tubules in skeletal muscle. AB - To examine the intracellular trafficking and translocation of GLUT4 in skeletal muscle, we have generated transgenic mouse lines that specifically express a GLUT4-EGFP (enhanced green fluorescent protein) fusion protein under the control of the human skeletal muscle actin promoter. These transgenic mice displayed EGFP fluorescence restricted to skeletal muscle and increased glucose tolerance characteristic of enhanced insulin sensitivity. The GLUT4-EGFP protein localized to the same intracellular compartment as the endogenous GLUT4 protein and underwent insulin- and exercise-stimulated translocation to both the sarcolemma and transverse-tubule membranes. Consistent with previous studies in adipocytes, overexpression of the syntaxin 4-binding Munc18c isoform, but not the related Munc18b isoform, in vivo specifically inhibited insulin-stimulated GLUT4-EGFP translocation. Surprisingly, however, Munc18c inhibited GLUT4 translocation to the transverse-tubule membrane without affecting translocation to the sarcolemma membrane. The ability of Munc18c to block GLUT4-EGFP translocation to the transverse-tubule membrane but not the sarcolemma membrane was consistent with substantially reduced levels of syntaxin 4 in the transverse-tubule membrane. Together, these data demonstrate that Munc18c specifically functions in the compartmentalized translocation of GLUT4 to the transverse-tubules in skeletal muscle. In addition, these results underscore the utility of this transgenic model to directly visualize GLUT4 translocation in skeletal muscle. PMID- 11054419 TI - A non-peptide functional antagonist of the CCR1 chemokine receptor is effective in rat heart transplant rejection. AB - Chemokines like RANTES appear to play a role in organ transplant rejection. Because RANTES is a potent agonist for the chemokine receptor CCR1, we examined whether the CCR1 receptor antagonist BX471 is efficacious in a rat heterotopic heart transplant rejection model. Treatment of animals with BX471 and a subtherapeutic dose of cyclosporin (2.5 mg/kg), which is by itself ineffective in prolonging transplant rejection, is much more efficacious in prolonging transplantation rejection than animals treated with either cyclosporin or BX471 alone. We have examined the mechanism of action of the CCR1 antagonist in in vitro flow assays over microvascular endothelium and have discovered that the antagonist blocks the firm adhesion of monocytes triggered by RANTES on inflamed endothelium. Together, these data demonstrate a significant role for CCR1 in allograft rejection. PMID- 11054420 TI - VLDL receptor fragments of different lengths bind to human rhinovirus HRV2 with different stoichiometry. An analysis of virus-receptor complexes by capillary electrophoresis. AB - The formation of complexes between the minor receptor group human rhinovirus HRV2 and two recombinant soluble receptor fragments derived from the human very low density lipoprotein receptor (VLDLR) and containing ligand-binding repeats 1-3 (MBP.VLDLR(1-3)) or 1-8 (MBP.VLDLR(1-8)) fused to the carboxyl terminus of the maltose-binding protein was analyzed by affinity capillary electrophoresis. At low molar ratios of receptor/virus, the peaks corresponding to substoichiometric complexes were broad indicating heterogeneity. When the receptors were present in molar excess with respect to the virus, the peaks were sharp, suggesting saturation of all binding sites. For the determination of the stoichiometry, constant amounts of receptor were incubated with increasing amounts of virus, and the peak areas corresponding to free receptor were measured and plotted versus total virus concentration. Extrapolation of the linear part of the resulting curve to zero concentration of free receptor enabled quantitation of the molar ratios of the components present in the complex. Using this method, we determined that about 60 molecules of MBP.VLDLR(1-3) but only about 30 molecules of MBP.VLDLR(1-8) were bound per virion. PMID- 11054421 TI - Akt activation protects hippocampal neurons from apoptosis by inhibiting transcriptional activity of p53. AB - Survival factors suppress apoptosis by activating the serine/threonine kinase Akt. To investigate the molecular mechanism underlying activated Akt's ability to protect neurons from hypoxia or nitric oxide (NO) toxicity, we focused on the apoptosis-related functions of p53 and caspases. We eliminated p53 by employing p53-deficient neurons and increased p53 by infection with recombinant adenovirus capable of transducing p53 expression, and we now show that p53 is implicated in the apoptosis induced by hypoxia or NO treatments of primary cultured hippocampal neurons. Although hypoxia and NO induced p53, treatment with insulin-like growth factor-1 significantly inhibited caspase-3-like activation, neuronal death and transcriptional activity of p53. These insulin-like growth factor-1 effects are prevented by wortmannin, a phosphatidylinositol 3-kinase inhibitor. Adenovirus mediated expression of activated-Akt kinase suppressed p53-dependent transcriptional activation of responsive genes such as Bax, suppressed caspase-3 like protease activity and suppressed neuronal cell death with no effect on the cellular accumulation and nuclear translocation of p53. In contrast, overexpression of kinase-defective Akt failed to suppress these same activities. These results suggest a mechanism where Akt kinase activation reduces p53's transcriptional activity that ultimately rescues neurons from hypoxia- or NO mediated cell death. PMID- 11054422 TI - The human CLN2 protein/tripeptidyl-peptidase I is a serine protease that autoactivates at acidic pH. AB - The CLN2 gene mutated in the fatal hereditary neurodegenerative disease late infantile neuronal ceroid lipofuscinosis encodes a lysosomal protease with tripeptidyl-peptidase I activity. To understand the enzymological properties of the protein, we purified and characterized C-terminal hexahistidine-tagged human CLN2p/tripeptidyl-peptidase I produced from insect cells transfected with a baculovirus vector. The N terminus of the secreted 66-kDa protein corresponds to residue 20 of the primary CLN2 gene translation product, indicating removal of a 19-residue signal peptide. The purified protein is enzymatically inactive; however, upon acidification, it is proteolytically processed and concomitantly acquires enzymatic activity. The N terminus of the final 46-kDa processed form (Leu196) corresponds to that of mature CLN2p/tripeptidyl-peptidase I purified from human brain. The activity of the mature enzyme is irreversibly inhibited by the serine esterase inhibitor diisopropyl fluorophosphate, which specifically and stoichiometrically reacts with CLN2p/tripeptidyl-peptidase I at Ser475, demonstrating that this residue represents the active site nucleophile. Expression of wild type and mutant proteins in CHO cells indicate that Ser475, Asp360, Asp517, but not His236 are essential for activity. These data indicate that the CLN2 gene product is synthesized as an inactive proenzyme that is autocatalytically converted to an active serine protease. PMID- 11054423 TI - F plasmid conjugative DNA transfer: the TraI helicase activity is essential for DNA strand transfer. AB - The product of the Escherichia coli F plasmid traI gene is required for DNA transfer via bacterial conjugation. This bifunctional protein catalyzes the unwinding of duplex DNA and is a sequence-specific DNA transesterase. The latter activity provides the site- and strand-specific nick required to initiate DNA transfer. To address the role of the TraI helicase activity in conjugative DNA transfer traI mutants were constructed and their function in DNA transfer was evaluated using genetic and biochemical methods. A traI deletion/insertion mutant was transfer-defective as expected. A traI C-terminal deletion that removed the helicase-associated motifs was also transfer-defective despite the fact that the region of traI encoding the transesterase activity was intact. Biochemical studies demonstrated that the N-terminal domain was sufficient to catalyze oriT dependent transesterase activity. Thus, a functional transesterase was not sufficient to support DNA transfer. Finally, a point mutant, TraI-K998M, that lacked detectable helicase activity was characterized. This protein catalyzed oriT-dependent transesterase activity in vitro and in vivo but failed to complement a traI deletion strain in conjugative DNA transfer assays. Thus, both the transesterase and helicase activities of TraI are essential for DNA strand transfer. PMID- 11054424 TI - A hybrid between Na+,K+-ATPase and H+,K+-ATPase is sensitive to palytoxin, ouabain, and SCH 28080. AB - Na(+),K(+)-ATPase is inhibited by cardiac glycosides such as ouabain, and palytoxin, which do not inhibit gastric H(+),K(+)-ATPase. Gastric H(+),K(+) ATPase is inhibited by SCH28080, which has no effect on Na(+),K(+)-ATPase. The goal of the current study was to identify amino acid sequences of the gastric proton-potassium pump that are involved in recognition of the pump-specific inhibitor SCH 28080. A chimeric polypeptide consisting of the rat sodium pump alpha3 subunit with the peptide Gln(905)-Val(930) of the gastric proton pump alpha subunit substituted in place of the original Asn(886)-Ala(911) sequence was expressed together with the gastric beta subunit in the yeast Saccharomyces cerevisiae. Yeast cells that express this subunit combination are sensitive to palytoxin, which interacts specifically with the sodium pump, and lose intracellular K(+) ions. The palytoxin-induced K(+) efflux is inhibited by the sodium pump-specific inhibitor ouabain and also by the gastric proton pump specific inhibitor SCH 28080. The IC(50) for SCH 28080 inhibition of palytoxin induced K(+) efflux is 14.3 +/- 2.4 microm, which is similar to the K(i) for SCH 28080 inhibition of ATP hydrolysis by the gastric H(+),K(+)-ATPase. In contrast, palytoxin-induced K(+) efflux from cells expressing either the native alpha3 and beta1 subunits of the sodium pump or the alpha3 subunit of the sodium pump together with the beta subunit of the gastric proton pump is inhibited by ouabain but not by SCH 28080. The acquisition of SCH 28080 sensitivity by the chimera indicates that the Gln(905)-Val(930) peptide of the gastric proton pump is likely to be involved in the interactions of the gastric proton-potassium pump with SCH 28080. PMID- 11054425 TI - A kinetic model for the metabolic interaction of two substrates at the active site of cytochrome P450 3A4. AB - In many cases, CYP3A4 exhibits unusual kinetic characteristics that result from the metabolism of multiple substrates that coexist at the active site. In the present study, we observed that alpha-naphthoflavone (alpha-NF) exhibited a differential effect on CYP3A4-mediated product formation as shown by an increase and decrease, respectively, of the carboxylic acid (P(2)) and omega-3 hydroxylated (P(1)) metabolites of losartan, while losartan was found to be an inhibitor of the formation of the 5,6-epoxide of alpha-NF. Thus, to address this problem, a kinetic model was developed on the assumption that CYP3A4 can accommodate two distinct and independent binding domains for the substrates within the active site, and the resulting velocity equations were employed to predict the kinetic parameters for all possible enzyme-substrate species. Our results indicate that the predicted values had a good fit with the experimental observations. Therefore, the kinetic constants can be used to adequately describe the nature of the metabolic interaction between the two substrates. Applications of the model provide some new insights into the mechanism of drug-drug interactions at the level of CYP3A4. PMID- 11054426 TI - Circular minichromosomes become highly recombinogenic in topoisomerase-deficient yeast cells. AB - In topoisomerase-deficient yeast cells, we have found that circular minichromosomes are present as broad distributions of multimeric forms, which consist of tandemly repeated copies of their monomeric sequences. This phenomenon selectively occurs in Deltatop1 cells, and is highly magnified in double mutant Deltatop1 top2-4 cells. No multimers are observed in single mutant top2-4 or Deltatop3 cells, or in Deltatop1 cells that express a plasmid-borne TOP1 gene. Interconversion among multimeric forms takes place rapidly in double mutant Deltatop1 top2-4 cells, and the multimeric distributions are readily reverted to the monomeric form when a plasmid-borne TOP1 gene is expressed from an inducible promoter. These observations are a new example of the interplay between DNA topology and genome stability, and suggest that the cell capacity to modulate DNA supercoiling is limited when DNA is organized in small topological domains. Yeast minichromosome multimerization provides an appropriate system in which to study mechanistic aspects of DNA recombination. PMID- 11054427 TI - Characterization of the biosynthesis of human immunodeficiency virus type 1 Env from infected T-cells and the effects of glucose trimming of Env on virion infectivity. AB - HIV (human immunodeficiency virus)-1 Env is displayed on the surface of infected cells and subsequently incorporated into virions, which is necessary for the initiation of a viral infection by recognition of the CD4 and the chemokine receptors (such as CCR5 or CXCR4) on the surface of new target cells. As a type 1 integral membrane glycoprotein, Env is cotranslationally translocated into the endoplasmic reticulum. In this report, we characterized the synthesis of Env, which did not occur at a constant rate but by translational/translocational pausing that has not previously been shown with a viral encoded glycoprotein. Overall translation was not impeded by the presence of the reducing agent dithiothreitol in vivo, although this did influence the cleavage of the precursor gp160 into its mature form, gp120. Env interacts transiently with resident components of the endoplasmic reticulum such as calnexin, which had maximal association at a 10-min post-translation. Addition of the glucosidase inhibitor, castanospermine, failed to significantly influence the association of Env with calnexin, consistent with the notion that calnexin recognizes components other than alpha-terminal glucose. Moreover, castanospermine treatment failed to affect the infectivity of virions. Taken together, this report demonstrates the existence of translational/translocational pausing for a viral glycoprotein and suggests that trimming of glucose from HIV-1 Env is not essential for the initiation of virus infection. PMID- 11054428 TI - NMR characterization of a DNA duplex containing the major acrolein-derived deoxyguanosine adduct gamma -OH-1,-N2-propano-2'-deoxyguanosine. AB - The environmental and endogenous mutagen acrolein reacts with cellular DNA to produce several isomeric 1,N(2)-propanodeoxyguanosine adducts. High resolution NMR spectroscopy was used to establish the structural features of the major acrolein-derived adduct, gamma-OH-1,N(2)-propano-2'-deoxyguanosine. In aqueous solution, this adduct was shown to assume a ring-closed form. In contrast, when gamma-OH-1,N(2)-propano-2'-deoxyguanosine pairs with dC at the center of an 11 mer oligodeoxynucleotide duplex, the exocyclic ring opens, enabling the modified base to participate in a standard Watson-Crick base pairing alignment. Analysis of the duplex spectra reveals a regular right-handed helical structure with all residues adopting an anti orientation around the glycosidic torsion angle and Watson-Crick alignments for all base pairs. We conclude from this study that formation of duplex DNA triggers the hydrolytic conversion of gamma-OH-1,N(2) propano-2'-deoxyguanosine to an open chain form, a structure that facilitates pairing with dC during DNA replication and accounts for the surprising lack of mutagenicity associated with this DNA adduct. PMID- 11054429 TI - Mismatch extension ability of yeast and human DNA polymerase eta. AB - DNA polymerase eta (Poleta) functions in error-free replication of UV-damaged DNA, and in vitro it efficiently bypasses a cis-syn T-T dimer by incorporating two adenines opposite the lesion. Steady state kinetic studies have shown that both yeast and human Poleta are low-fidelity enzymes, and they misincorporate nucleotides with a frequency of 10(-2)-10(-3) on both undamaged and T-T dimer containing DNA templates. To better understand the role of Poleta in error-free translesion DNA synthesis, here we examine the ability of Poleta to extend from base mismatches. We find that both yeast and human Poleta extend from mismatched base pairs with a frequency of approximately 10(-3) relative to matched base pairs. In the absence of efficient extension of mismatched primer termini, the ensuing dissociation of Poleta from DNA may favor the excision of mismatched nucleotides by a proofreading exonuclease. Thus, we expect DNA synthesis by Poleta to be more accurate than that predicted from the fidelity of nucleotide incorporation alone. PMID- 11054430 TI - The nitric oxide congener nitrite inhibits myeloperoxidase/H2O2/ Cl- -mediated modification of low density lipoprotein. AB - Nitric oxide, a pivotal molecule in vascular homeostasis, is converted under aerobic conditions to nitrite. Recent studies have shown that myeloperoxidase (MPO), an abundant heme protein released by activated leukocytes, can oxidize nitrite (NO(2-)) to a radical species, most likely nitrogen dioxide. Furthermore, hypochlorous acid (HOCl), the major strong oxidant generated by MPO in the presence of physiological concentrations of chloride ions, can also react with nitrite, forming the reactive intermediate nitryl chloride. Since MPO and MPO derived HOCl, as well as reactive nitrogen species, have been implicated in the pathogenesis of atherosclerosis through oxidative modification of low density lipoprotein (LDL), we investigated the effects of physiological concentrations of nitrite (12.5-200 microm) on MPO-mediated modification of LDL in the absence and presence of physiological chloride concentrations. Interestingly, nitrite concentrations as low as 12.5 and 25 microm significantly decreased MPO/H2O2)/Cl- -induced modification of apoB lysine residues, formation of N-chloramines, and increases in the relative electrophoretic mobility of LDL. In contrast, none of these markers of LDL atherogenic modification were affected by the MPO/H2O2/NO2-) system. Furthermore, experiments using ascorbate (12.5-200 microm) and the tyrosine analogue 4-hydroxyphenylacetic acid (12.5-200 microm), which are both substrates of MPO, indicated that nitrite inhibits MPO-mediated LDL modifications by trapping the enzyme in its inactive compound II form. These data offer a novel mechanism for a potential antiatherogenic effect of the nitric oxide congener nitrite. PMID- 11054431 TI - MYCN expression in neuroblastoma: A mixed message? PMID- 11054432 TI - Prospective trial of the herbal supplement PC-SPES in patients with progressive prostate cancer. AB - PURPOSE: PC-SPES is an herbal supplement for which there are anecdotal reports of anti-prostate cancer activity. This phase II study was undertaken to assess the efficacy and toxicity of PC-SPES in prostate cancer patients. PATIENTS AND METHODS: Thirty-three patients with androgen-dependent prostate cancer (ADPCa) and 37 patients with androgen-independent prostate cancer (AIPCa) were treated with PC-SPES at a dose of nine capsules daily. Clinical outcome was assessed with serial serum prostate-specific androgen (PSA) level measurement and imaging studies. RESULTS: One hundred percent of ADPCa patients experienced a PSA decline of >/= 80%, with a median duration of 57+ weeks. No patient has developed PSA progression. Thirty-one patients (97%) had declines of testosterone to the anorchid range. Two ADPCa patients had positive bone scans; both improved. One patient with a bladder mass measurable on computed tomography scan experienced disappearance of this mass. Nineteen (54%) of 35 AIPCa patients had a PSA decline of >/= 50%, including eight (50%) of 16 patients who had received prior ketoconazole therapy. Median time to PSA progression was 16 weeks (range, 2 to 69+ weeks). Of 25 patients with positive bone scans, two had improvement, seven had stable disease, 11 had progressive disease, and five did not have a repeat bone scan because of PSA progression. Severe toxicities included thromboembolic events (n = 3) and allergic reactions (n = 3). Other frequent toxicities included gynecomastia/gynecodynia, leg cramps, and grade 1 or 2 diarrhea. CONCLUSION: PC SPES seems to have activity in the treatment of both ADPCa and AIPCa and has acceptable toxicity. Further study is required to determine whether its effects exceed those expected with estrogen therapy. PMID- 11054433 TI - MYCN expression is not prognostic of adverse outcome in advanced-stage neuroblastoma with nonamplified MYCN. AB - PURPOSE: The clinical significance of MYCN expression in children with neuroblastoma (NB) remains controversial. To determine the prognostic significance of MYCN expression in the absence of MYCN amplification, we analyzed MYCN mRNA and protein expression in tumors from 69 patients. PATIENTS AND METHODS: Sixty-nine NB tumor samples with nonamplified MYCN from patients with stage C or D disease were obtained from the Pediatric Oncology Group Neuroblastoma Tumor Bank. MYCN mRNA was analyzed using a real-time reverse transcriptase polymerase chain reaction assay, and MYCN protein was examined by Western blot analyses. RESULTS: The estimated 5-year event-free survival (EFS) and survival (S) rates plus SE for the cohort were 57% +/- 17% and 60% +/- 16%, respectively. Infants younger than 1 year had significantly higher rates of EFS and S than children >/= 1 year of age (P =.003 and P <.001, respectively); patients with stage C disease had better outcome than those with stage D NB (P <.001); and patients with hyperdiploid tumors had better outcome than those with diploid NB (P <.001). Surprisingly, outcome was slightly better for patients with high versus low levels of MYCN mRNA expression (4-year S, 70% +/- 13% v 50% +/- 16%; P =.290), and for patients with tumors that expressed MYCN protein (4-year S, 73% +/- 19% v 53% +/- 15%, respectively; P =.171). CONCLUSION: High levels of MYCN expression are not prognostic of adverse outcome in patients with advanced stage NB with nonamplified MYCN. A trend associating high levels of MYCN expression with improved outcome was observed. PMID- 11054434 TI - Prospective validation of renal function-based carboplatin dosing in children with cancer: A United Kingdom Children's Cancer Study Group Trial. AB - PURPOSE: Carboplatin dosing in adults with cancer is based on renal function. The purpose of the current study was to validate a previously developed pediatric carboplatin-dosing formula. PATIENTS AND METHODS: Thirty-eight pediatric patients were randomized to receive a carboplatin dose calculated according to surface area or a renal function-based dosing formula. On the next course of therapy, the alternative dosing method was used for each patient. Carboplatin pharmacokinetics (based on free plasma platinum concentrations) were measured after both courses. RESULTS: The mean observed areas under the carboplatin concentration-versus-time curve (AUCs) after renal function- and surface area-based dosing were 98% and 95% of the target AUCs, respectively. The variation in the observed AUC was significantly less after renal function-based dosing (F test, P =.02), such that 74% of courses had an observed AUC within +/- 20% of the target value, versus 49% for courses after dosing according to surface area. Only one of 22 courses at the center with the most experience with renal function-based dosing was associated with an AUC outside +/- 20% of the target value, versus nine of 22 courses after surface area-based dosing in the same center. There was a relationship (r(2) =.71) between carboplatin AUC and thrombocytopenia in 10 neuroblastoma patients treated with a combination of carboplatin, vincristine, etoposide, and cyclophosphamide. CONCLUSION: Renal function-based carboplatin dosing in children results in more consistent drug exposure than surface area-based drug administration. PMID- 11054435 TI - Durable remission after aggressive chemotherapy for very late post-kidney transplant lymphoproliferation: A report of 16 cases observed in a single center. AB - PURPOSE: Posttransplant lymphoproliferative diseases (PTLDs) represent a group of potentially lethal lymphoid proliferations that may complicate the course of solid organ transplantation. Although early-onset PTLDs frequently have a favorable outcome, late-onset PTLDs behave more alike aggressive lymphoma. We report a monocentric retrospective study that focused on PTLDs occurring later than 1 year after kidney transplantation (very late-onset PTLDs) to define their incidence, clinical presentation, pathologic features, and outcome. We particularly emphasized the follow-up of patients treated with conventional chemotherapy. PATIENTS AND METHODS: The medical histories of all patients who developed very late-onset PTLD in our institution were reviewed, and diagnostic biopsy materials were retrospectively studied. RESULTS: Very late-onset PTLDs were diagnosed in 16 (1.1%) of 1,421 patients. Mean (+/- SD) time to tumor onset was 103.93 +/- 70.88 months. Most tumors were Epstein-Barr virus-related monomorphic large-cell PTLDs of B phenotype. Ten patients received conventional chemotherapy (cyclophosphamide, doxorubicin, vincristine, and prednisone regimen). Two of them died within 2 months, two achieved partial remission, and six achieved definitive complete remission. Overall median survival time was 13 months and rose to 27 months in the treated group. The main cause of mortality was sepsis. None of the treated patients experienced rejection despite withdrawal of immunosuppressive treatment. CONCLUSION: Despite characteristics of aggressive lymphoma, very late-onset PTLDs after renal transplantation may respond to conventional chemotherapy. However, because a high rate of infectious complications occurred, new therapeutic strategies, such as combinations of anti CD20 monoclonal antibodies and lower doses of chemotherapy, are warranted. PMID- 11054436 TI - Role of a doxorubicin-containing regimen in relapsed and resistant lymphomas: an 8-year follow-up study of EPOCH. AB - PURPOSE: Curative up-front regimens for non-Hodgkin's lymphomas contain doxorubicin, vincristine, and cyclophosphamide, whereas salvage regimens generally contain non-cross-resistant agents. We hypothesized that up-front agents may be highly effective for salvage and developed an infusional regimen based on in vitro evidence of increased efficacy. PATIENTS AND METHODS: A prospective phase II study of etoposide, vincristine, and doxorubicin over 96 hours with bolus cyclophosphamide and oral prednisone (EPOCH) was performed in 131 patients with relapsed or resistant lymphoma. RESULTS: Seventy-nine percent of patients had aggressive histologies, 46% were considered high risk by the International Prognostic Index, and 34% had resistant disease. Eighty-eight percent of patients had received at least four of the agents in EPOCH, and 94% had received doxorubicin. In 125 assessable patients, 29 (24%) achieved complete responses and 60 (50%) achieved partial responses. Among 42 patients with resistant disease, 57% responded, and in 28 patients with relapsed aggressive de novo lymphomas, 89% responded with 54% complete responses. With a median follow up of 76 months, the overall and event-free survivals (EFS) were 17.5 and 7 months, respectively. In 33 patients with sensitive aggressive disease who did not receive stem-cell transplantation, EFS was 19% at 36 months. Toxicity was primarily hematologic, with an 18% incidence of febrile neutropenia. No clinically significant cardiac toxicity was observed, despite no maximum cumulative doxorubicin dose. CONCLUSION: EPOCH is highly effective in patients who had previously received most/all of the same drugs and produces durable remissions in curable subtypes. Salvage regimens need not contain non-cross resistant agents, and infusional schedules may partially reverse drug resistance and reduce toxicity. PMID- 11054437 TI - High-dose chemotherapy with autologous hematopoietic stem-cell transplantation for advanced soft tissue sarcoma in adults. AB - PURPOSE: Patients with metastatic or locally advanced, unresectable soft tissue sarcoma (ASTS) are seldom curable, with 5-year survival rates of less than 10% in all large series. The role of high-dose chemotherapy (HDCT) with hematopoietic stem-cell support in this disease is not established. PATIENTS AND METHODS: Between 1988 and 1994, 30 patients with ASTS who responded to a standard chemotherapy regimen were included in a prospective pilot study of HDCT as consolidation therapy using ifosfamide (12 g/m(2)), etoposide (800 mg/m(2)), and cisplatin (200 mg/m(2)) (VIC). RESULTS: The median duration of grade 4 neutropenia and thrombocytopenia was 14 and 10 days, respectively. Nineteen patients (63%) experienced grade 1 or higher renal toxicity. All eight patients in complete remission (CR) before HDCT were still in CR at day 60. Of the 22 patients in partial remission (PR) or with a minor response to conventional chemotherapy, CR, PR, and stable disease were achieved in four (18%), three (13%), and 12 patients (54%), respectively, by day 60, while three patients (14%) progressed. With a median follow-up of 94 months, overall and progression-free survival rates at 5 years after HDCT were 23% and 21%, respectively. Patients in CR before HDCT had a significantly superior 5-year overall survival rate compared with other patients (75% v 5%; P: =.001). CONCLUSION: Despite the toxicity of the VIC regimen, a high survival rate was observed in HDCT-treated patients who were in CR after conventional chemotherapy. A phase III randomized trial is required to establish the role of HDCT in ASTS. PMID- 11054438 TI - Assessment of methods for tissue-based detection of the HER-2/neu alteration in human breast cancer: a direct comparison of fluorescence in situ hybridization and immunohistochemistry. AB - PURPOSE: To compare the efficacy of fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) in detecting the HER-2/neu alteration in human breast cancer. PATIENTS AND METHODS: Unselected stage I, II, and III breast cancer patients (N = 900) were tested for HER-2/neu gene amplification by FISH in paraffin-embedded, formalin-fixed archival material. Of these samples, 856 were tested for HER-2/neu overexpression by non-antigen-retrieval IHC with the polyclonal antibody R60, the sensitivity and specificity of which was preliminarily compared with the United States Food and Drug Administration approved HercepTest (Dako Corp, Carpinteria, CA). Patient survival was analyzed in relation to the presence of the HER-2/neu alteration as determined by these two methodologies. RESULTS: A total of 189 (21%) of 900 patients were positive by FISH and 147 (17.2%) of 856 were positive by IHC. This discrepancy is consistent with expected loss of IHC sensitivity associated with tissue fixation/embedding. The HercepTest did not improve sensitivity and introduced false positives. Comparison of R60-based IHC with FISH demonstrates that patient survival is associated progressively to gene amplification level as determined by FISH, whereas for IHC an association is found only in the highest (3+) immunostaining group. Among FISH-negative tumors, 45 (6.6%) of 678 were IHC-positive, with a survival probability similar to that of FISH-negative/IHC-negative cases; FISH positive/IHC-negative patients have a survival probability similar to that of FISH-positive/IHC-positive cases. CONCLUSION: IHC does not consistently discriminate patients likely to have a poor prognosis, whereas FISH provides superior prognostic information in segregating high-risk from lower-risk beast cancers. HER-2/neu protein overexpression in the absence of gene amplification occurs infrequently in breast cancer, in which case, patient outcome is similar to that of patients without the alteration. PMID- 11054439 TI - Physician compliance with warfarin prophylaxis for central venous catheters in patients with solid tumors. AB - PURPOSE: There is an established benefit of prophylactic warfarin in cancer patients with central venous catheters. This study assessed the compliance rate of prophylactic low-dose warfarin prescription in cancer patients with central venous catheters at a single institution. PATIENTS AND METHODS: Oncology patients with central venous catheters were identified by a retrospective chart review. Information retrieved included whether prophylactic warfarin had been prescribed and whether the patient had suffered a thrombotic or bleeding event. After the initial chart review, physicians were notified of the benefits of warfarin prophylaxis, and subsequently, a physician-independent mechanism of prescribing prophylactic warfarin was instituted. After each of these interventions, we retrospectively reviewed a further two cohorts of patients to assess compliance with warfarin prophylaxis. RESULTS: During the baseline study, only 10% of patients were prescribed prophylactic warfarin. After physician notification, the compliance rate increased to only 20% (P =.3). After instituting the physician independent mechanism of prescribing prophylactic warfarin, the compliance rate increased to 95% (P <.001). The rate of catheter-related thrombosis was 11% for patients who were prescribed warfarin compared with 21% in those who were not anticoagulated (P =.2). CONCLUSION: At our institution, the rate of prescribing prophylactic warfarin was low in this patient population, and there was a reluctance of treating physicians to change their prescribing practice. Mechanisms exist to improve the rate of anticoagulant prophylaxis in this clinical setting. We recommend that institutions review their rate of compliance with prophylactic anticoagulation for patients with central venous catheters and solid tumors. PMID- 11054440 TI - Dietary fat intake and endogenous sex steroid hormone levels in postmenopausal women. AB - PURPOSE: To examine the relationship between plasma levels of reproductive sex steroid hormones in postmenopausal women and their reported fat intake. METHODS: We measured plasma sex steroid hormones levels in plasma collected in 1989 and 1990 from 381 healthy postmenopausal women. For each woman, we measured fat intake in 1986 and 1990 by a food-frequency questionnaire. The cross-sectional associations between the percentage of energy from total and specific types of dietary fat intake and plasma hormone levels were assessed by linear regression, controlling for energy intake, obesity, and protein intake. RESULTS: The plasma estradiol level was 4.3% lower (95% confidence limits, -8.3%, -0.2%) for a substitution of 5% of energy from fat intake for an equivalent amount of energy from carbohydrate when adjusted for obesity and other covariates. Estradiol was also inversely associated with all other fat types except trans fat; the inverse associations with vegetable fat and marine omega-3 fats were statistically significant. CONCLUSION: We observed an inverse association between total fat intake averaged over 4 to 5 years and estradiol levels. This result is inconsistent with the hypothesis that fat intake predisposes to breast cancer risk by raising endogenous estrogen levels. PMID- 11054441 TI - Phase I dose-finding and pharmacokinetic study of paclitaxel and carboplatin with oral valspodar in patients with advanced solid tumors. AB - PURPOSE: To evaluate the maximum-tolerated dose (MTD), dose-limiting toxicities (DLTs), and pharmacokinetic (PK) profile of paclitaxel and carboplatin when administered every 3 weeks with the oral semisynthetic cyclosporine analog valspodar (PSC 833), an inhibitor of P-glycoprotein function. PATIENTS AND METHODS: Fifty-eight patients were treated with escalating doses of paclitaxel ranging from 54 to 94.5 mg/m(2) and carboplatin area under the plasma concentration versus time curve (AUC) ranging from 6 to 9 mg.min/mL, every 21 days. The dose of valspodar was fixed at 5 mg/kg every 6 hours for a total of 12 doses from day 0 to day 3. The MTD was determined for the following two groups: (1) previously treated patients, where paclitaxel and carboplatin doses were escalated; and (2) chemotherapy-naive patients, where paclitaxel dose was escalated and carboplatin AUC was fixed at 6 mg.min/mL. PK studies of paclitaxel and carboplatin were performed on day 1 of course 1. RESULTS: Fifty-eight patients were treated with 186 courses of paclitaxel, carboplatin, and valspodar. Neutropenia, thrombocytopenia, and hepatic transaminase elevations were DLTs. In previously treated patients, no DLTs occurred at the first dose level (paclitaxel 54 mg/m(2) and carboplatin AUC 6 mg.min/mL). However, one of 12, two of six, two of four, four of 11, and two of five patients experienced DLTs at doses of paclitaxel (mg/m(2))/carboplatin AUC (mg.min/mL) of 67.5/6, 81/6, 94.5/6, 67. 5/7.5, and 67.5/9, respectively. In chemotherapy-naive patients, one of 17 developed DLT at paclitaxel 81 mg/m(2) and carboplatin AUC 6 mg/mL.min. There was prolongation of the terminal phase of paclitaxel elimination as evidenced by an increased time that plasma paclitaxel concentration was >/= 0.05 micromol/L, ranging from 16.6 +/- 6.7 hours to 41.5 +/- 9.8 hours for paclitaxel doses of 54.5 mg/m(2) to 94.5 mg/m(2), respectively. CONCLUSION: The recommended phase II dose in chemotherapy-naive patients is paclitaxel 81 mg/m(2), carboplatin AUC 6 mg.min/mL, and valspodar 5 mg/kg every 6 hours. In previously treated patients, the recommended phase II dose is paclitaxel 67.5 mg/m(2), carboplatin AUC 6 mg.min/mL, and valspodar 5 mg/kg every 6 hours. The acceptable toxicity profile supports the rationale for performing disease-directed evaluations of paclitaxel, carboplatin and valspodar on the schedule evaluated in this study. PMID- 11054442 TI - Meropenem versus ceftazidime in the treatment of cancer patients with febrile neutropenia: a randomized, double-blind trial. AB - PURPOSE: To compare meropenem, a carbapenem antibiotic, with ceftazidime for the empirical treatment of patients with febrile neutropenia. PATIENTS AND METHODS: A prospective, double-blind, randomized clinical trial was conducted at medical centers in North America and the Netherlands. A total of 411 cancer patients (196 treated with meropenem and 215 treated with ceftazidime), who had 471 episodes of fever, participated in the trial. For each neutropenic episode, patients were allocated at random to receive intravenous administration of meropenem (1 g every 8 hours) or ceftazidime (2 g every 8 hours). Treatment could be modified at any time. Key end points were clinical and bacteriologic outcomes, eradication of infecting organism, and adverse events. RESULTS: The rate of successful clinical response at the end of therapy was significantly higher for patients treated with meropenem than for those on ceftazidime for all episodes (54% v 44%, respectively) and for episodes of fever of unknown origin (62% v 46%, respectively), but differences between groups were not statistically significant for clinically defined or microbiologically defined infections. Meropenem was significantly more effective than ceftazidime in severely neutropenic (Asn, Gln409-->Pro and Gly447-->Ser, two long-to-short loop replacement mutations, Gly23-Ala24-Asp25-Gly26-Ala27-Trp28- Val29-Ser30-->Asn-Pro-Pro (23-30 replacement) and Asp297-Ser298-Glu299-Ala300 Val301-->Ala-G ly-Ala (297-301 replacement) and one deletion mutation removing Glu439, Thr440 and Ser441 (Delta439-441), all based on amino acid sequence alignments, were made to improve Aspergillus awamori glucoamylase thermostability. The first and second single-residue mutations were designed to introduce a potential N:-glycosylation site and to restrict backbone bond rotation, respectively, and therefore to decrease entropy during protein unfolding. The third single-residue mutation was made to decrease flexibility and increase O:-glycosylation in the already highly O:-glycosylated belt region that extends around the globular catalytic domain. The 23-30 replacement mutation was designed to eliminate a very thermolabile extended loop on the catalytic domain surface and to bring the remainder of this region closer to the rest of the catalytic domain, therefore preventing it from unfolding. The 297-301 replacement mutant GA was made to understand the function of the random coil region between alpha-helices 9 and 10. Delta439-441 was constructed to decrease belt flexibility. All six mutations increased glucoamylase thermostability without significantly changing enzyme kinetic properties, with the 23-30 replacement mutation increasing the activation free energy for thermoinactivation by about 4 kJ/mol, which leads to a 4 degrees C increase in operating temperature at constant thermostability. PMID- 11054461 TI - Apo(a)-kringle IV-type 6: expression in Escherichia coli, purification and in vitro refolding. AB - Lipoprotein (a) [Lp(a)] belongs to the class of highly thrombo-atherogenic lipoproteins. The assembly of Lp(a) from LDL and the specific apo(a) glycoprotein takes place extracellularly in a two-step process. First, an unstable complex is formed between LDL and apo(a) due to the interaction of the unique kringle (K) IV type 6 (T6) in apo(a) with amino groups on LDL, and in the second step this complex is stabilized by a disulfide bond between apo(a) KIV-T9 and apoB(100). In order to understand this process better, we overexpressed and purified apo(a) KIV T6 in Escherichia coli. Recombinant KIV-T6 was expressed as a His-tag fusion protein under control of the T7 promoter in BL21 (DE3) strain. After one-step purification by affinity chromatography the yield was 7 mg/l of bacterial suspension. Expressed fusion apo(a) KIV-T6 was insoluble in physiological buffers and it also lacked the characteristic kringle structure. After refolding using a specific procedure, high-resolution (1)H-NMR spectroscopy revealed kringle structure-specific signals. Refolded KIV-T6 bound to Lys-Sepharose with a significantly lower affinity than recombinant apo(a) (EC(50) with epsilon-ACA 0.47 mM versus 2-11 mM). In competition experiments a 1000-fold molar excess of KIV-T6 was needed to reach 60% inhibition of Lp(a) assembly. PMID- 11054462 TI - Clusterin protein diversity in the primate eye. AB - PURPOSE: The clusterin gene encodes a multi-functional protein that has been identified in different tissues, including a number of different eye tissues, primarily in the mouse and to a much lesser extent in humans. Clusterin has been implicated in a number of cellular processes such as lipid transport, membrane integrity, apoptosis, and neurodegeneration, all of which could be important to the biology of the eye. In the current communication, we provide data that confirms the expression of clusterin in a number of different human eye tissues and establishes the expression profile of this gene in monkey derived eye tissues. The issue that we sought to examine is whether a broad profile of clusterin expression in the eye is consistent in primates (monkey and human). METHODS: The majority of our study was done using monkey eye tissues. Where possible, we have used human tissues in order to confirm published findings. Northern and western analysis was performed using tissues derived from monkey eyes. In situ hybridization and immunochemistry were carried out on human eye sections. RESULTS: Clusterin mRNA is expressed in primate lens, cornea, limbus, sclera, orbital muscle, ciliary body, retina, RPE/choroid, and RPE cells in culture. Western analysis revealed that two major groups of clusterin exist in the eye, a high molecular weight group (>100 kDa) and a second group consisting of at least five clusterin species that are all approximately 80 kDa. Analysis of conditioned media from RPE cells cultured on permeable supports suggests that different forms of clusterin display alternative patterns of secretion. CONCLUSIONS: Clusterin is expressed in a broad range of eye tissues in both human and monkey, suggesting that this is a characteristic feature in primates. We demonstrate for the first time that a diverse number of clusterin isoforms were observed in monkey eye tissues by western analysis. Meanwhile, the molecular size of clusterin mRNA detected in the array of tissues are identical in size, suggesting that the nature of the diversity in clusterin forms is due to post translational modifications. In addition, new insights were made in defining clusterin expression in ciliary body, cornea, and the retinal pigment epithelium. PMID- 11054463 TI - The fibrillating atrial myocardium. What can the detection of wave breaks tell us? PMID- 11054464 TI - Reperfusion injury and ischemic preconditioning: two sides of a coin? PMID- 11054465 TI - Acquired delayed rectifier channelopathies: how heart disease and antiarrhythmic drugs mimic potentially-lethal congenital cardiac disorders. PMID- 11054466 TI - Sarcoplasmic reticulum calcium release function and FK binding proteins in heart failure: another piece of a complex jigsaw. PMID- 11054467 TI - Kinin B(1) receptors and the cardiovascular system: regulation of expression and function. AB - Kinins are important peptide mediators of a diverse range of physiological and pathological functions of the cardiovascular system. The kinin peptides exert their effects by selective activation of two distinct G-protein coupled receptors termed B(1) and B(2). The principal kinin peptides involved in the acute regulation of cardiovascular function during normal physiology are bradykinin (BK) and Lys-BK which produce their effects via activation of B(2) receptors. The B(1) receptor is activated by the des-Arg(9)kinin metabolites namely des-Arg(9)BK and Lys-des-Arg(9)BK, the synthesis of which are increased during inflammation. The B(1) receptor, which is not constitutively expressed, is induced in various pathologies relating to inflammation. Recent investigations into the molecular mechanisms of B(1) receptor induction and their distribution and function in the cardiovascular system have shown that following an inflammatory stimulus the B(1) receptor is induced and may play an important role in modulation of cardiovascular function. This review summarises recent studies on B(1) receptor expression and function in the cardiovascular system and discusses the role of these receptors in regulation of circulatory homeostasis and their potential as therapeutic targets. PMID- 11054468 TI - beta(1) and beta(2)-adrenergic receptor subtype effects in German shepherd dogs with inherited lethal ventricular arrhythmias. AB - OBJECTIVE: Delayed afterdepolarization-induced triggered activity originating in ventricular myocardium is a mechanism for some age-dependent, inherited ventricular tachycardias in a colony of German shepherd dogs. METHODS: We used standard microelectrode techniques to study beta-adrenergic receptor subtype modulation of the triggered activity in anteroseptal left ventricular myocardium from eleven of these dogs and seven unafflicted, age-matched German shepherd controls. RESULTS: During sustained stimulation at cycle lengths of 300-4000 ms, 10(-9)-10(-7) M isoproterenol concentration-dependently shortened action potential duration (APD) to 90% repolarization more in myocardium from afflicted than from unafflicted dogs. This shortening was prevented by a beta(1)-blocker CGP20712A (10(-7) M) while a beta(2)-blocker ICI118551 (10(-7) M) did not modify the effect of isoproterenol in either group. The beta(2)-agonist zinterol 10(-8) 10(-6) M had no effect on APD. Stimulation at a cycle length of 250 ms in the presence of 10(-7) M isoproterenol induced more triggered AP in myocardium from afflicted than unafflicted dogs. beta(1)-Blockade completely eliminated, while beta(2)-blockade facilitated, and the beta(2)-agonist zinterol did not induce triggered activity in the two groups. CONCLUSION: Isoproterenol effects on APD and triggered activity in the myocardium of dogs with inherited arrhythmias are due primarily to an abnormality of beta(1)-adrenoceptor mediated signaling that is subject to beta(2)-adrenergic modulation. PMID- 11054469 TI - Dynamics of wavelets and their role in atrial fibrillation in the isolated sheep heart. AB - BACKGROUND: The multiple wavelet hypothesis is the most commonly accepted mechanism underlying atrial fibrillation (AF). However, high frequency periodic activity has recently been suggested to underlie atrial fibrillation in the isolated sheep heart. We hypothesized that in this model, multiple wavelets during AF are generated by fibrillatory conduction away from periodic sources and by themselves may not be essential for AF maintenance. METHODS AND RESULTS: We have used a new method of phase mapping that enables identification of phase singularities (PSs), which flank individual wavelets during sustained AF. The approach enabled characterization of the initiation, termination, and lifespan of wavelets formed as a result of wavebreaks, which are created by the interaction of wave fronts with functional and anatomical obstacles in their path. AF was induced in six Langendorff-perfused sheep hearts in the presence of acetylcholine. High resolution video imaging was utilized in the presence of a voltage sensitive dye; two-dimensional phase maps were constructed from optical recordings. The major results were as follows: (1) the critical inter-PS/wavelet distance for the formation of rotors was 4 mm, (2) the spatial distribution of wavelets/PSs was non-random. (3) the lifespan of PSs/wavelets was short; 98% of PSs/wavelets existed for < 1 rotation, and (4) the mean number of waves that entered our mapping field (15.7 +/- 1.6) exceeded the mean number of waves that exited it (9.7 +/- 1.5; P < 0.001). CONCLUSIONS: Our results strongly suggest that multiple wavelets may result from breakup of high frequency organized waves in the isolated Langendorff-perfused sheep heart, and as such are not a robust mechanism for the maintenance of AF in our model. PMID- 11054470 TI - Intra-uterine growth retardation results in increased cardiac arrhythmias and raised diastolic blood pressure in adult rats. AB - OBJECTIVES: Epidemiological evidence in humans suggests that intrauterine growth retardation is associated with an increased risk of hypertension and coronary heart disease in later life. To begin to understand the mechanisms involved, we developed and exploited a rat model of intrauterine growth retardation to assess predisposition to arrhythmias and resting blood pressure levels at defined ages from 4 to 18 months. METHODS: Isolated working heart experiments were carried out on rats that had been subjected to intrauterine growth retardation by prenatal protein deprivation and age-matched male Wistar controls to measure susceptibility to wall stress-induced arrhythmias. In addition, resting systolic and diastolic blood pressures were measured in conscious rats via an indwelling arterial catheter. RESULTS: Hearts from intrauterine growth retarded animals showed significantly more ventricular premature beats and more episodes of ventricular tachycardia at all ages examined (4, 9 and 18 months), and at 4 and 18 months, a reduction in coronary blood flow. Diastolic pressure was significantly raised by intrauterine growth retardation in both groups examined (4 and 9 months). CONCLUSIONS: Protein malnutrition during the intrauterine period results in profound intrauterine growth retardation that is associated with a raised diastolic blood pressure and an increased predisposition to cardiac arrhythmias in later life. These results are consistent with epidemiological observations made in human populations, and as similar pathophysiological changes may operate in both situations, intrauterine protein deprivation may be a useful model to help define some of the mechanisms involved. PMID- 11054471 TI - Activity of the Na(+)/H(+) exchanger is critical to reperfusion damage and preconditioning in the isolated rat heart. AB - OBJECTIVE: Removal of protons from the heart during ischemia and/or reperfusion by the cardiac Na(+)/H(+) exchanger (NHE1) leads to Na(+) entry; this causes Ca(2+) influx and is thought to contribute to ischemic and/or reperfusion damage. The extent to which Na(+) enters during ischemia as opposed to reperfusion is disputed and has important implications for the therapeutic use of NHE1 inhibitors as protection against ischemic damage. Preconditioning has recently been proposed to inhibit NHE1 during reperfusion. The objective of the present study was to determine the activity of NHE1 during ischemia, reperfusion and following preconditioning. METHODS: The experiments were on isolated perfused rat hearts in which left ventricular developed pressure (LVDP) and intracellular sodium and pH were measured. RESULTS: LVDP following 30 min of ischemia recovered to 14+/-3% of pre-ischemic level. Application of the NHE1 inhibitor HOE 642 during ischemia and reperfusion improved recovery of LVDP to 77+/-9%. When HOE was applied at the moment of reperfusion the recovery of LVDP was reduced to 54+/ 6%. To overcome possible delays in the delivery of HOE, the drug was applied at 28 min of ischemia; under these conditions recovery of LVDP (71+/-7%) was not significantly different to HOE throughout ischemia and reperfusion. HOE had no effect on the recovery of preconditioned hearts. NHE1 activity was assessed by the [Na(+)](i) and pH(i) changes in response to brief exposure to Na(+) lactate (NaL). In control hearts, activity of NHE1 caused a pH(i) recovery of 0. 034+/ 0.007 pH units and was associated with a [Na(+)](i) rise of 7. 5+/-0.5 mmol/l. After 5-min reperfusion following ischemia, NaL application caused a pH(i) recovery (0.046+/-0.007) and a larger [Na(+)](i) rise (15.8+/-0.6 mmol/l). After 5-min reperfusion of preconditioned hearts, NaL application caused a smaller recovery pH recovery (0.013+/-0.002) and a smaller [Na(+)](i) rise (4.2+/-0.5 mmol/l). CONCLUSIONS: These findings suggest that NHE1 is activated in early reperfusion after ischemia but inhibited during early reperfusion in preconditioned hearts. Overall our results point to a critical period of activity of NHE1 in early reperfusion which is inhibited by preconditioning. PMID- 11054472 TI - Localisation and functional significance of ryanodine receptors during beta adrenoceptor stimulation in the guinea-pig sino-atrial node. AB - OBJECTIVE: Recent evidence shows that calcium released from the sarcoplasmic reticulum (SR) plays an important role in the regulation of heart rate. The aim of this study was to investigate the subcellular distribution of ryanodine receptors in the guinea-pig sino-atrial (SA) node and to determine their functional role in the regulation of pacemaker frequency in response to beta adrenoceptor stimulation. METHODS: Monoclonal antibodies raised against the cardiac ryanodine receptor were used with confocal microscopy to investigate ryanodine receptor distribution in single guinea-pig SA node cells. The functional role of ryanodine receptors was investigated in both multicellular SA node/atrial preparations and in single SA node cells. RESULTS: Ryanodine receptor labelling was observed in all SA node cells studied and showed both subsarcolemmal and intracellular staining. In the latter, labelling appeared as transverse bands with a regular periodicity of approximately 2 microm. This interval resembled that of the expected sarcomere spacing but did not, however, depend on the presence of transverse tubules. The bands of ryanodine receptors appeared to be located in the region of the Z lines, based on co-distribution studies with antibodies to alpha-actinin, myomesin and binding sites for phalloidin. Functional studies on single SA node cells showed that application of ryanodine (2 micromol/l) reduced the rate of firing of spontaneous action potentials (measured using the perforated patch clamp technique) and this was associated with changes in action potential characteristics. Ryanodine also significantly decreased the positive chronotropic actions of isoprenaline in both multicellular and single cell preparations. In single cells exposed to 100 nmol/l isoprenaline, ryanodine caused a decrease in the rate of firing and this was associated with a decrease in the amplitude of the measured calcium transients. CONCLUSIONS: These findings are the first to show immunocytochemical evidence for the presence and organisation of ryanodine receptor calcium release channels in mammalian SA node cells. This study also provides evidence of a role for ryanodine sensitive sites in the beta-adrenergic modulation of heart rate in this species. PMID- 11054473 TI - Arrhythmogenic activity of cardiac muscle in pulmonary veins of the dog: implication for the genesis of atrial fibrillation. AB - OBJECTIVE: Pulmonary veins are important foci of ectopic beats to initiate paroxysmal atrial fibrillation. The purpose of this study were to investigate the electrophysiological characteristics of excitable cells in canine pulmonary veins obtained from healthy and chronic rapid atrial pacing dogs and their responses to cardioactive agents. METHODS: Transmembrane action potentials (APs) were recorded from multiple sites of pulmonary veins isolated from 17 healthy dogs and 14 dogs with chronic (6-8 weeks) rapid atrial pacing (780 bpm). RESULTS: In normal superfusate, several types of electrical activities were identified, including silent electrical activity, fast response APs driven by electrical stimulation, and spontaneous fast or slow response APs (with or without early afterdepolarizations). The incidences of AP with an early afterdepolarization (93% versus 41%) was greater in chronic pacing dogs. The spontaneous activities were depressed by beta-adrenoceptor blocker, calcium channel blocker, adenosine and acetylcholine. High frequency (>8 Hz) irregular rhythms occurred spontaneously or were induced by cardioactive agents or electrical stimuli. The incidence of spontaneously occurring tachyarrhythmias was much higher in preparations from chronic pacing dogs (93%) than from control (12%). The tachyarrhythmias were suppressed by sodium channel blocker, potassium channel blocker or magnesium. CONCLUSIONS: Pulmonary veins have arrhythmogenic ability through spontaneous activities or high-frequency irregular rhythms. The higher incidence of spontaneously occurring high-frequency irregular rhythms in chronic rapid atrial pacing dogs may account for the increased risk of atrial fibrillation in these dogs. PMID- 11054474 TI - Comitogenic effect of catecholamines on rat cardiac fibroblasts in culture. AB - OBJECTIVE: We studied the ability of norepinephrine and of other catecholamines to affect the proliferation of cardiac fibroblasts isolated from adult rat hearts. Furthermore, we investigated the possible adrenergic receptor involved in this process. METHODS: Norepinephrine (NE), phenylephrine (PE), isoproterenol (ISO), forskolin (FO), epidermal growth factor (EGF), platelet-derived growth factor AA (PDGF-AA) and specific inhibitors of the alpha(1)-, alpha(2)-, beta(1)- and beta(2)-adrenoceptors and of the protein kinase A (PKA) were applied to cardiac fibroblasts in culture. Cell number was measured by use of a Coulter Counter. Activation of the cAMP response element binding protein (CREB) was measured by Western blotting and subsequent use of a phospho-specific antibody. Activation of the p42- and the p44-mitogen activated protein kinase (p42/p44(MAPK)) was assessed by detection of phosphorylation shifts and by incorporation of 32P-labelled phosphate into myelin basic protein. RESULTS: Fibroblasts isolated from hearts of adult rats were grown in 10% serum-containing media which induced an increase in cell number by 94%. After 48 h, treatment with 10 microM NE caused an even greater increase in cell number by 222%, i.e. another 128% (comitogenic effect). In contrast, NE alone had no effect on the growth of serum-deprived cells. EGF and PDGF-AA did not replace serum as the basic mitogen. After addition of NE to proliferating cells under serum conditions, there was a rapid, time-dependent significant activation of the p42/p44(MAPK) and of CREB for up to 60 and 120 min, respectively. In both cases, the maximum of activation was reached after 5 min. Application of FO (0.1-20 microM) caused a strong activation of CREB, while no increase in the phosphorylation of the p42/p44(MAPK) was detected. Treatment with 20 microM FO led to an identical increase in cell number as application of NE. Specific blockade of PKA with RpcAMPS prevented the activation of CREB and also the comitogenic effect of FO as well as of NE. The alpha- and beta-adrenergic receptor blocker carvedilol (10 microM) normalized all NE-induced effects. Prazosin and yohimbine, inhibitors of alpha(1)- and alpha(2) adrenoceptor activation, respectively, did not influence the NE-evoked increase in cell number. In contrast, the non-selective beta-adrenoceptor blocker propranolol (1 microM) completely suppressed the comitogenic effect of NE. A similar effect was obtained with the specific beta(2)-adrenoceptor blocker ICI 118,551 (5 microM), while the beta(1)-adrenoceptor blocker metoprolol did not influence the increase in cell number. CONCLUSIONS: NE elicits a comitogenic effect on cultured rat cardiac fibroblasts which is prevented by beta(2) adrenergic blockade. The activation of CREB contributes to the increase in proliferation. The p42/p44(MAPK) which was also found to be activated by NE might as well be involved in the regulation of the comitogenic effect. PMID- 11054475 TI - Molecular heterogeneity of protein kinase C expression in human ventricle. AB - OBJECTIVE: Although activation of protein kinase C (PKC) modulates the function of normal cardiac myocytes and likely plays a role in the pathogenesis of cardiomyopathic disease states, the molecular basis of PKC expression in human ventricle has not been examined in detail. METHODS: We have performed Western analysis and immunohistochemistry on explanted human cardiac tissue from nondiseased and diseased specimens using isoform-specific antibodies directed against all known PKC isozymes. RESULTS: In homogenates from left and right ventricle, all isoforms except PKC-gamma and theta were detected by immunoblotting, with confirmation using a second antibody directed against a different epitope when possible. For PKC-betaII, delta, and epsilon, data indicated that these isoforms were variably phosphorylated in vivo, resulting in multiple bands during immunoblotting. Because of potential antibody cross reactivity, reverse transcriptase polymerase chain reaction (RT-PCR) was performed which confirmed expression of PKC-alpha, betaI, and zeta. Immunohistochemistry demonstrated that all isoforms detected in ventricular homogenate by Western analysis could be localized to cardiac myocytes. From a methodologic standpoint, significant degradation of PKC isoforms could be demonstrated when samples were either frozen or allowed to remain at room temperature, compared to immediate subcellular fractionation. CONCLUSIONS: These findings indicate that the PKC expression in human ventricular myocytes is remarkably diverse, with multiple conventional, novel, and atypical isoforms present, and highlight the importance of sample preparation in comparative studies of PKC isoform expression. PMID- 11054476 TI - Pacing-induced heart failure causes a reduction of delayed rectifier potassium currents along with decreases in calcium and transient outward currents in rabbit ventricle. AB - OBJECTIVE: Heart failure in patients and in animal models is associated with action potential prolongation of the ventricular myocytes. Changes in several membrane currents have been already demonstrated to underlie this prolongation. However, information on the two components (I(Kr) and I(Ks)) of the delayed rectifier potassium current (I(K)) in rapid pacing induced heart failure is lacking. METHODS AND RESULTS: Action potentials and whole-cell currents, I(K), I(to1), I(K1), and I(Ca-L) were recorded in apical myocytes of left ventricle from 10 rabbits subjected to left ventricular pacing at 350-380 beats/min for 3-4 weeks and 10 controls with sham operation. Action potential duration at 90% repolarization (APD(90)) was prolonged in myocytes from failing hearts compared to controls at both cycle lengths of 333 and 1000 ms. Both E-4031-sensitive and resistant components of I(K) (I(Kr), I(Ks)) in myocytes from failing hearts were significantly less than those of control hearts; tail current densities of I(Kr) and I(Ks) following depolarization to +50 mV were 0.62+/-0.05 vs. 0.96+/-0.12 pA/pF (P<0.05), and 0.27+/-0.08 vs. 0.52+/-0.08 pA/pF (P<0.05), respectively. There was no significant difference between control and failing myocytes in the voltage- and time-dependence of activation of total I(K), I(Kr) and I(Ks). The peak of L-type Ca(2+) current (I(Ca-L)) was significantly reduced in myocytes from failing hearts (at +10 mV, -9.29+/-0.52 vs. -12.28+/-1.63 pA/pF, P<0.05), as was the Ca(2+)-independent transient outward current (I(to1); at +40 mV, 4.8+/ 0.9 vs. 9.6+/-1.3 pA/pF, P<0.05). Steady state I-V curve for I(K1) was similar in myocytes from failing and control hearts. CONCLUSIONS: Decrease of I(K) (both I(Kr) and I(Ks)) in addition to reduced I(to1), may underly action potential prolongation at physiological cycle length and thereby contribute to arrhythmogenesis in heart failure. PMID- 11054477 TI - cGMP-dependent protein kinase mediates stimulation of L-type calcium current by cGMP in rabbit atrial cells. AB - OBJECTIVES: cGMP has been shown to exert both stimulatory and inhibitory effects on cardiac L-type calcium current (I(Ca)). The physiological role of cGMP in regulation of cardiac activity is still controversial. cGMP may be of importance in regulation of I(Ca) in atrial cells. The present study was focused on the role of cGMP in the modulation of I(Ca) in rabbit atrial cells. METHODS: Enzymatically isolated adult rabbit atrial cells were used to measure I(Ca) using whole cell voltage clamp. Expressed levels of cGMP-dependent protein kinase (PKG) were determined by Western blotting using PKG specific antibody in homogenates from atrial and ventricular cells. RESULTS: Nitrosoglutathione (GSNO), a nitric oxide donor that stimulates soluble guanylyl-cyclase to elevate cGMP levels increased I(Ca) while soluble G-cyclase inhibitors, ODQ or methylene blue inhibited I(Ca). Intracellular application of 8BrcGMP increased I(Ca) and blocked the inhibitory effect of methylene blue. KT-5823, an inhibitor of PKG inhibited I(Ca) and the stimulatory effect of GSNO was completely blocked ODQ or KT-5823. Inhibition of cAMP dependent protein kinase (PKA) by the 6-22 peptide completely blocked the stimulation of I(Ca) by the beta-agonist isoproterenol but not by GSNO. The potency of isoproterenol to stimulate I(Ca) was very high for atrial cells (EC(50) 2.4+/-0.6 nM) and only 100 nM isoproterenol was required to stimulate I(Ca) maximally (21.4+/-0.7 pA/pF) to a level (23.8+/-1.6 pA/pF) achieved with the inclusion of 100 microM cAMP in the pipette solution. GSNO produced an additive effect on I(Ca) already stimulated by either 10 microM isobutylmethylxanthine (phosphodiesterase inhibitor) or a low concentration (1 nM) isoproterenol but failed to produce any effect on I(Ca) maximally stimulated by 100 nM isoproterenol. Inhibition of PKG by KT-5823 significantly decreased the efficacy of isoproterenol and the maximal I(Ca) achieved with 100 nM isoproterenol was decreased to 8.2+/-0.6 pA/pF in the presence of KT-5823. Western blot analysis showed much higher expression of PKG in atrial cells compared to ventricular cells. CONCLUSIONS: These findings suggest that stimulatory effects of cGMP on I(Ca) in rabbit atrial cells are likely to be mediated via PKG dependent phosphorylation of calcium channels or associated proteins and that the effects of cGMP are not antagonistic to cAMP. PKG is highly expressed in atrial cells and PKG dependent phosphorylation may be necessary for maintaining basal I(Ca) and fully stimulating I(Ca) by beta-adrenergic activation in atrial cells. PMID- 11054478 TI - Altered interaction of FKBP12.6 with ryanodine receptor as a cause of abnormal Ca(2+) release in heart failure. AB - OBJECTIVE: Little information is available as to the Ca(2+) release function of the sarcoplasmic reticulum (SR) in heart failure. We assessed whether the alteration in this function in heart failure is related to a change in the role of FK binding protein (FKBP), which is tightly coupled with the cardiac ryanodine receptor (RyR) and recently identified as a modulatory protein acting to stabilize the gating function of RyR. METHODS: SR vesicles were isolated from dog LV muscles [normal (N), n=6; heart failure induced by 3-weeks pacing (HF), n=6]. The time course of the SR Ca(2+) release was continuously monitored using a stopped-flow apparatus, and [3H]ryanodine-binding and [3H]dihydro-FK506-binding assays were also performed. RESULTS: FK506, which specifically binds to FKBP12.6 and dissociates it from RyR, decreased the polylysine-induced enhancement of [3H]ryanodine-binding by 38% in N (P<0.05) but it had no effect in HF. In HF, the rate constant for the polylysine-induced Ca(2+) release from the SR was 61% smaller than in N. FK506 decreased the rate constant for the polylysine-induced Ca(2+) release by 67% in N (P<0.05) but had no effect in HF. The [3H]dihydro FK506-binding assay revealed that the number (B(max)) of FKBPs was decreased by 83% in HF (P<0.05), while the K(d) value was unchanged. FK506 did not significantly change SR Ca(2+.)-ATPase activity in either N or HF. CONCLUSIONS: In HF, the number of FKBPs showed a tremendous decrease; this may underlie the RyR-channel instability and the impairment of the Ca(2+) release function of RyR seen in the failing heart. PMID- 11054479 TI - Chronic therapy with an ET(A/B) receptor antagonist in conscious dogs during progression of congestive heart failure. Intracellular Ca(2+) regulation and nitric oxide mediated coronary relaxation. AB - BACKGROUND: Although it is known that endothelin (ET-1) is elevated in heart failure (HF), it remains unclear if chronic ET(A/B) receptor antagonism affects the progression of HF, particularly by affecting coronary vasoactivity and left ventricular (LV) diastolic function. METHODS: We examined the effects of an ET(A/B) receptor antagonist, L-753,037 (oral bid for 6 weeks, n=7), and vehicle (n=8) in conscious dogs with previously implanted aortic, coronary sinus and left atrial catheters, LV pressure gauge, aortic flow probe, LV dimension crystals and pacers. RESULTS: Baseline hemodynamics were similar in the two groups. During the development of rapid pacing-induced HF, treatment with the ET(A/B) antagonist significantly reduced total peripheral resistance and increased cardiac output compared to vehicle. After 2 weeks of pacing, LV diastolic function (tau) was improved (P<0.05) in the ET(A/B) antagonist group (+6+/-2 ms) compared to the vehicle group (+12+/-2 ms). In addition, ET(A/B) antagonist treatment attenuated the increase in mean left atrial pressure and LV end-diastolic pressure that occurred during heart failure in vehicle-treated animals. However, LV systolic function (LV dP/dt, fractional shortening and Vcfc) neither at rest nor in response to dobutamine was altered by ET(A/B) antagonist treatment. Also, ET(A/B) antagonist treatment did not affect the progressive increases in LV dimension. After 6 weeks of pacing, maximal Ca(2+) transport in isolated cardiac sarcoplasmic reticulum (SR) was reduced (P<0.02) in the vehicle-treated compared to the ET(A/B) antagonist-treated dogs (1.34+/-0.09 vs. 1.60+/-0.06 micromol/mg/min, respectively). The improvement in SR function in the ET(A/B) antagonist-treated dogs was associated with a significant attenuation of the reduction in protein expression of SERCA2a and calsequestrin observed in the vehicle-treated dogs. Coronary arteries isolated from the dogs treated with the ET(A/B) antagonist exhibited enhanced (P<0.01) coronary endothelium-dependent relaxation compared to the vehicle group, while coronary responses to an NO donor were identical in the two groups. Plasma NO levels in the coronary sinus during the late stage of HF were higher (P<0.05) in the ET(A/B) antagonist group (40+/-2 microM) compared to the vehicle group (18+/-2 microM). CONCLUSIONS: We conclude that in conscious dogs during the development of HF induced by rapid pacing, chronic inhibition of ET(A/B) receptors does not affect resting myocardial contractile function nor reserve, but reduces vascular resistance and improves LV diastolic function. After 6 weeks of pacing, the reduction in intracellular Ca(2+) regulation by the SR is also attenuated, and endothelium-dependent coronary relaxation is improved, which appears to be related to the preservation of coronary NO levels. PMID- 11054480 TI - Contraction in cardiac endothelial cells contributes to changes in capillary dimensions following ischaemia and reperfusion. AB - OBJECTIVE: Ischaemia followed by reperfusion brings about a reduction in cardiac capillary cross-sectional dimensions which is consistent with constriction. The aim of this study was to test the hypothesis that the reduction in cardiac capillary dimensions that occurs in ischaemia and reperfusion is caused by endothelial cell contraction and that modulating the endothelial cell contractile apparatus reduces microvascular reperfusion injury. METHODS: In isolated rat hearts we used phalloidin to stabilise the endothelial actin filaments in order to prevent the dimensional changes during ischaemia. The changes in endothelial cell dimensions were quantified by measuring whole capillary and luminal cross sectional areas, abluminal and luminal membrane lengths. We have also used resin casts of the coronary vasculature coupled with scanning electron microscopy to examine the structural changes along the length of the capillaries in ischaemia reperfusion. RESULTS: We found that the reduction in capillary dimensions was prevented by the addition of phalloidin and, in the resin casts, that ischaemia reperfusion cause focal narrowings along the capillaries which are consistent with constriction. CONCLUSIONS: (1) The endothelial contractile apparatus is involved in the reduction in cross-sectional dimensions. (2) This implies that the capillary bed may have a greater role in the local control of flow than was previously thought and that modulation of the actomyosin contractile system in cardiac capillary endothelial cells may be useful in reducing 'no reflow' injury which results from reperfusion. PMID- 11054481 TI - Arrhythmogenic action of endothelin-1. PMID- 11054482 TI - Brain ischemia and reperfusion: molecular mechanisms of neuronal injury. AB - Brain ischemia and reperfusion engage multiple independently-fatal terminal pathways involving loss of membrane integrity in partitioning ions, progressive proteolysis, and inability to check these processes because of loss of general translation competence and reduced survival signal-transduction. Ischemia results in rapid loss of high-energy phosphate compounds and generalized depolarization, which induces release of glutamate and, in selectively vulnerable neurons (SVNs), opening of both voltage-dependent and glutamate-regulated calcium channels. This allows a large increase in cytosolic Ca(2+) associated with activation of mu calpain, calcineurin, and phospholipases with consequent proteolysis of calpain substrates (including spectrin and eIF4G), activation of NOS and potentially of Bad, and accumulation of free arachidonic acid, which can induce depletion of Ca(2+) from the ER lumen. A kinase that shuts off translation initiation by phosphorylating the alpha-subunit of eukaryotic initiation factor-2 (eIF2alpha) is activated either by adenosine degradation products or depletion of ER lumenal Ca(2+). Early during reperfusion, oxidative metabolism of arachidonate causes a burst of excess oxygen radicals, iron is released from storage proteins by superoxide-mediated reduction, and NO is generated. These events result in peroxynitrite generation, inappropriate protein nitrosylation, and lipid peroxidation, which ultrastructurally appears to principally damage the plasmalemma of SVNs. The initial recovery of ATP supports very rapid eIF2alpha phosphorylation that in SVNs is prolonged and associated with a major reduction in protein synthesis. High catecholamine levels induced by the ischemic episode itself and/or drug administration down-regulate insulin secretion and induce inhibition of growth-factor receptor tyrosine kinase activity, effects associated with down-regulation of survival signal-transduction through the Ras pathway. Caspase activation occurs during the early hours of reperfusion following mitochondrial release of caspase 9 and cytochrome c. The SVNs find themselves with substantial membrane damage, calpain-mediated proteolytic degradation of eIF4G and cytoskeletal proteins, altered translation initiation mechanisms that substantially reduce total protein synthesis and impose major alterations in message selection, down-regulated survival signal-transduction, and caspase activation. This picture argues powerfully that, for therapy of brain ischemia and reperfusion, the concept of single drug intervention (which has characterized the approaches of basic research, the pharmaceutical industry, and clinical trials) cannot be effective. Although rigorous study of multi-drug protocols is very demanding, effective therapy is likely to require (1) peptide growth factors for early activation of survival-signaling pathways and recovery of translation competence, (2) inhibition of lipid peroxidation, (3) inhibition of calpain, and (4) caspase inhibition. Examination of such protocols will require not only characterization of functional and histopathologic outcome, but also study of biochemical markers of the injury processes to establish the role of each drug. PMID- 11054483 TI - Fatigue and basal ganglia. AB - Fatigue is a common symptom in neurology and occurs in the diseases of the central and peripheral nervous system. In order to understand the mechanism of fatigue, it is important to distinguish symptoms of peripheral neuromuscular fatigue from the symptoms of physical and mental fatigue characteristic of disorders like Parkinson's disease or multiple sclerosis. We have introduced and defined the concept of central fatigue for the latter disorders. We have further proposed, with supportive neuropathological data, that central fatigue may occur due to a failure in the integration of the limbic input and the motor functions within the basal ganglia affecting the striatal-thalamic-frontal cortical system. PMID- 11054484 TI - Long-term effect of IFNbeta1b treatment on the spontaneous and induced expression of IL-10 and TGFbeta1 in MS patients. AB - Interferon-beta (IFNbeta) is an effective treatment that lessens the frequency and severity of exacerbations in relapsing-remitting multiple sclerosis (RRMS). The mechanism of action of IFNbeta1b may be by upregulating antiinflammatory cytokines levels. We studied the effect of IFNbeta1b treatment on the in vivo gene expression and protein synthesis of two immunosuppressive cytokines, IL-10 and TGFbeta1, and its persistence with chronic therapy. Peripheral blood samples were obtained from 16 patients before and after 3, 6 and 12 months of IFNbeta1b treatment. Eleven patients did not have any clinical relapse, whereas the other five each had one clinical exacerbation during the study. We employed a highly sensitive RT-PCR technique to study the spontaneous gene expression of IL-10 and TGFbeta1. Protein concentration in serum and in culture supernatants from mitogen stimulated cells were measured by ELISA. In the group of patients who remained clinically stable during the study, IL-10 mRNA levels decreased significantly after 6 months of treatment to normalize at 1 year of therapy as compare with the initial values. In the five patients who relapsed, mRNA IL-10 levels were significantly diminished at 3, 6, and 12 months of therapy. IL-10 serum levels did not vary significantly in any group of patients during the study. Treatment did not modulate mRNA or serum levels of TGFbeta1 at any time period in the group of stable patients. However, in the five patients who relapsed, TGFbeta1 mRNA significantly decreased at 6 and 12 months of therapy. IFNbeta1b treatment was unable to restore the initial low mitogen-induced production of IL-10; only after 1 year of therapy was a slight increase observed. Cytokine therapy did not affect the mitogen-induced production of TGFbeta1. We can conclude that chronic administration of IFNbeta1b does not result in an upregulation of IL-10 and TGFbeta1. PMID- 11054485 TI - Asymmetrical temporal lobe atrophy with massive neuronal inclusions in multiple system atrophy. AB - This report concerns a rare association of asymmetrical temporal lobe atrophy with multiple system atrophy (MSA). A 53-year-old Japanese woman developed cerebellar ataxia and parkinsonism and was diagnosed as olivopontocerebellar atrophy (OPCA). This patient showed forgetfulness and subsequent disorientation even in the early stage of the disease. She fell into a decorticate state at the age of 64, and died a year later. The autopsy showed MSA with asymmetrical atrophy of temporal lobes, intraneuronal globular inclusions mostly confined to the hippocampus, amygdaloid nucleus, and most abundant in the granule cells in the dentate fascia. These inclusions were intensely argyrophilic and expressed marked immunoreactivity to ubiquitin, but not to neurofilament (NF), tau and paired helical filaments (PHF). Ultrastructurally, they were composed of scattered short filamentous structures of 15 to 30 nm in diameter, ribosome-like granules, mitochondria and lipofuscin. The lack of immunoreactivity against tau, NF and PHF suggests that the inclusions are distinct from Pick bodies. To our knowledge, MSA in association with asymmetrical temporal lobe atrophy with the present neuronal inclusions has not been reported. This case is distinct from MSA combined with atypical Pick's disease in the distribution and immunohistochemical properties of neuronal inclusions, and may present a new variant of MSA since the neuronal inclusions are similar, in many respects, to those of neuronal inclusions reported in MSA. Globular inclusions are also discussed in variants of Pick's disease, amyotrophic lateral sclerosis and Alzheimer's disease. PMID- 11054486 TI - Cognitive dysfunction in individuals with myasthenia gravis. AB - In the present study we administered a battery of cognitive measures that examined attention, response fluency, information processing, and verbal and visual learning and retention to 28 individuals with generalized myasthenia gravis (MG) and 18 demographically similar control subjects. Results revealed that MG patients performed significantly more poorly than control subjects on the measures of response fluency, information processing and most measures of verbal and visual learning. Significant group differences were not evident on the measure of attention span or on the indices of retention of information. Cognitive performances of the MG group were not related to mood disturbance, disease duration, or daily dose of prednisone. While these results suggest central involvement in MG, previous studies have not provided evidence that MG antibodies bind to central nicotinic receptors. Possible alternative mechanisms underlying cognitive dysfunction in MG are discussed. PMID- 11054487 TI - Continuous pulse oximetry in acute hemiparetic stroke. AB - BACKGROUND AND PURPOSE: Hypoxemia can adversely affect ischemic brain tissue in laboratory animals. The aim of this study was to assess the value of early continuous monitoring with pulse oximetry in detecting arterial oxygen desaturations in patients with acute hemiparetic stroke, and the effects of oxygen administration. METHODS: Over a period of 6 months 49 consecutive patients with acute hemiparetic stroke of /=75 years old). As for sm, in the group <75 years, old tumours have been the leading cause of mortality since 1985/87, but in the older age group CV-nsm has been more than twice TU-nsm. By 1991/93 in comparison with 1982/84, CV deaths have fallen by 6% (-28% in the age group <75 years, +3% in the age group >/=75 years), while TU deaths have grown by 17% (+3% in the age group <75 years, +45% in the age group >/=75 years). Considering all age groups, by 1991/93 the absolute number of CV-d (239.241) was much greater than the number of TU-d (151.908); overall, almost 70% of CV-d and 40% of TU-d took place in the older age group. For the near future, the rapid aging of the Italian population (from 1982/84 to 1991/93 there was a 40% increment in the population older than 75 years) is a relevant variable to take into account. Thus, despite the 'reassuring' fall in CV-sm and nsm, cardiovascular diseases are expected to remain the major cause of death and physical disability in adults. PMID- 11054505 TI - Matrix metalloproteinase-9 and tissue inhibitor metalloproteinase-1 levels in essential hypertension. Relationship to left ventricular mass and anti hypertensive therapy. AB - To test the hypothesis that the activity of enzymes degrading the extracellular matrix in hypertensive patients are abnormal, and that the treatment of hypertension will normalise these abnormalities, we measured the serum levels of metalloproteinase MMP-9, and its inhibitor, tissue metalloproteinase inhibitor (TIMP-1). Thirty-two patients with untreated hypertension (BP 168/96) had significantly lower levels of both MMP-9 and TIMP-1 when compared to 24 matched normotensive controls (BP 123/80) (P<0.001). There was no significant correlation between MMP-9 and TIMP-1 levels (P>0.2). In the patients, there were no significant correlations observed between left ventricular mass, Doppler V(E)/V(A) ratio (an index of diastolic function), blood pressure, left ventricular mass index and either MMP-9 or TIMP-1 levels (all P=NS). Levels of MMP-9 and TIMP-1 were not significantly altered after 2 months of antihypertensive treatment of 29 patients despite mean blood pressure falling from 170/96 to 143/85 mmHg (P<0.001). Correspondingly, there were also no significant alterations in indices of diastolic function and left ventricular mass. Our study suggests that the proteolytic activities of MMP-9 and TIMP-l are depressed in hypertensive patients and were not significantly affected by short term antihypertensive treatment. The relationship between collagen metabolism in hypertensive subjects, especially in those with cardiac hypertrophy, and the effects of treatment needs to be further explored in larger trials over a longer period of time. PMID- 11054506 TI - Differential coronary hemodynamics and left ventricular contractility in patients with syndrome X. AB - The relationship between coronary hemodynamics and left ventricular contractility was studied in 20 patients with syndrome X. Among them, 10 patients with a resting left ventricular ejection fraction (LVEF, by radionuclide method) equal to or greater than the mean value of the whole group (58%) were defined as having relative increased left ventricular contractility (group H), and another 10 patients with relatively normal contractility (50%/=1.3+/-0.072) were highly suspicious for florid endocarditis. TTE and TEE were true positive in two and in six patients, whereas false positives were seen in two and in four patients. Scintigraphy was positive in four of the five patients with the false negative TTE and negative in the three false positive TEE. Vice versa, TEE was positive in the two patients with false negative scintigraphy. CONCLUSIONS: Immunoscintigraphy with 99mTc-Fab' fragments in combination with TEE improves diagnostic accuracy compared with TTE/TEE in patients with subacute infective endocarditis. PMID- 11054511 TI - The influence of treatment of hypercholesterolemic patients with simvastatin on plasma chemotactic activity and adherence of neutrophils. AB - BACKGROUND: there is some evidence to indicate that statins may affect the function of immune and inflammatory cells. This study investigates the influence of short term treatment with simvastatin on plasma chemotactic activity and adherence of polymorphonuclear neutrophils in hypercholesterolemic patients. METHODS AND RESULTS: 20 hypercholesterolemic patients (250-400 mg/dl) were given simvastatin (20 mg daily for 12 weeks). Peripheral blood samples were taken before and after 4 and 12 weeks of the therapy. The percentage of neutrophils adhering to plastic surface coated with albumin was significantly higher when cells were incubated with plasma obtained after 12 weeks of treatment with simvastatin in comparison with plasma collected before the therapy (unstimulated neutrophils: 5.945+/-0.475% vs. 8.155+/-0.96%, P=0.0477, stimulated neutrophils: 39.09+/-4.540% vs. 29.18+/-3.702%, P=0.032). There was a significant negative correlation between adherence of stimulated neutrophils and total cholesterol levels ((r)=-0.2796, 95% CI -0. 4999 to -0.02526, r(2)=0.07817, P=0.032). Migration of neutrophils towards plasma obtained after 12 weeks of treatment with simvastatin was significantly higher than towards plasma collected before the therapy (7.038+/-1.127 vs. 4.505+/-0 618 P=0.0475). CONCLUSION: treatment of hypercholesterolemic patients with simvastatin increases the chemotactic activity of plasma and augments the adherence of human neutrophils. PMID- 11054512 TI - Does atrial overdrive pacing prevent paroxysmal atrial fibrillation in paced patients? AB - The role of atrial overdrive pacing for the suppression of paroxysmal atrial fibrillation remains unclear. To investigate this we have performed a randomised study evaluating the role of an increased atrial base rate in suppressing this arrhythmia in patients implanted with a permanent pacemaker (Chorum ELA) for sick sinus syndrome with previous documented paroxysmal atrial fibrillation. Twenty seven patients (mean age, 69; 15 female) were randomised to two 3-month single blinded crossover periods of DDDR pacing. The pacemaker was set with a base rate of 60 bpm (normal) during one period and at 10 bpm (overdrive) above the average heart rate during the other, mean (S.D.) 75+/-7 beats/min (range, 70-96). The fallback algorithm of the pacemaker was activated to record the number and duration of paroxysmal atrial fibrillation episodes. During the overdrive period there was a significant increase in the total duration of atrial pacing (normal 60+/-26% vs. overdrive 72+/-28%, P<0.001). However there was no significant difference in the number of paroxysmal atrial fibrillation episodes (normal 43+/ 109 vs. overdrive 43+/-106, P=ns), or their total duration (normal 42+/-108 h vs. overdrive 99+/-254 h, P=ns). In conclusion, atrial overdrive pacing, achieved by increasing the atrial base rate, has no incremental benefit in the suppression of paroxysmal atrial fibrillation when compared to rate responsive pacing with a base rate of 60 bpm. PMID- 11054513 TI - Has the outlook improved for amifostine as a clinical radioprotector? AB - Amifostine has recently been approved for clinical radiotherapy as a protector against irradiation-induced xerostomia. It is our aim to review the outlook for using amifostine as a general clinical radioprotector. Protection against X-rays is mainly obtained by the scavenging of free radicals. The degree of protection is therefore highly dependent on oxygen tension, with protection factors ranging from 1 to 3. Maximal protection is observed at physiological levels of oxygenation. A great variability in protection has also been observed between different normal tissues. Some tissue, like brain, is not protected while salivary glands and bone marrow may exhibit a three-fold increase in radiation tolerance. Amifostine is dephosphorylized to its active metabolite by a process involving alkaline phosphatase. Due to lower levels of alkaline phosphatase in tumor vessels, amifostine is marketed as a selective protector of normal tissue and not tumors. However, the preclinical investigations concerning the selectivity of amifostine are controversial and the clinical studies are sparse and do not have the power to evaluate the influence of amifostine on the therapeutic index. CONCLUSION: based on the present knowledge amifostine should only be used in experimental protocols and not in routine practice. PMID- 11054514 TI - Continuous hyperfractionated accelerated radiotherapy with/without mitomycin C in head and neck cancers. AB - BACKGROUND AND PURPOSE: Radiation therapy is often the primary treatment for advanced cases of head and neck cancers not considered suitable for radical surgery. In these cases locoregional tumour control rates are low and has warranted innovative treatment modifications, such as altered fractionation schedules and combination with chemotherapy. PATIENTS AND METHODS: From October 1990 to December 1997, 239 patients with squamous cell cancers originating in the head and neck region were randomized to one of three treatment options. Standard therapy consisting of conventional fractionation with 70 Gy in 7 weeks in 35 fractions (CF). The second treatment option consisted of a continuous hyperfractionated accelerated radiotherapy delivering a total dose of 55.3 Gy in 33 fractions over 17 consecutive days (V-CHART). The third study arm had identical fractionation and dose as the above accelerated treatment, with the additional administration of 20 mg/m(2) mitomycin C (MMC) on day 5 of treatment (V-CHART+MMC). RESULTS: Main toxicity resulted from accelerated fractionation in confluent mucositis (Grade 3-4 in 95%) requiring nasogastral tube feeding, analgetics and antiphlogistics in the majority of cases. Haematological toxicity Grade 3-4 was seen after MMC administration in 18%. MMC administration did not influence mucosal reaction. Overall duration of mucositis was not different in the three treatment groups. Loco-regional tumour control was 31% after CF, 32% after V-CHART and 48% after V-CHART+MMC, respectively (P<0.05). Overall crude survival was 24% after CF, 31% following V-CHART and 41% after V-CHART+MMC, respectively (P<0.05). Median follow up was 48 months (assessment performed in February 1999). CONCLUSION: Following shortening overall treatment time from 7 weeks to 17 consecutive days and dose of radiotherapy from 70 to 55.3 Gy the results in the radiotherapy only treated patients are identical. A significant improvement regarding local tumour control and survival was seen following administration of MMC to the accelerated fractionated treatment. PMID- 11054515 TI - Double-blinded, randomized, placebo-controlled study to evaluate the effectiveness of sulphasalazine in preventing acute gastrointestinal complications due to radiotherapy. AB - BACKGROUND AND PURPOSE: Acute radiation-induced diarrhea occurs in approximately 80% of the patients receiving pelvic radiotherapy. It is caused by gastrointestinal irritation and inflammation. Eicosanoids are thought to be one of the mechanisms of this. Sulphasalazine is an inhibitor of their synthesis in the mucosa. This randomized clinical trial was undertaken to evaluate its effect in preventing acute radiation enteritis (ARE). MATERIALS AND METHODS: Prospectively, 87 patients receiving pelvic radiotherapy were randomized, in a double-blind fashion. Two tablets twice daily of sulphasalazine (500 mg) or placebo were administered orally. Patients were evaluated weekly according to diarrhea grading for the primary study endpoint and according to late effect of normal tissue-subjective objective management analytic (LENT-SOMA) criteria for the secondary endpoint during irradiation. RESULTS: Groups did not differ for age, gender, tumour site or irradiation procedure. Diarrhea occurred in 55 and 86% of the sulphasalazine and placebo groups, respectively (P=0.001). Gastrointestinal toxicity was seen in 80 and 93% of the sulphasalazine and placebo groups according to the maximum LENT-SOMA score (P=0.07). According to the maximum LENT-SOMA score between the two groups, significant differences in favor of sulphasalazine were found for each week. CONCLUSION: Sulphasalazine (2 g/day) was found to be effective in decreasing the symptoms of ARE. PMID- 11054516 TI - The effect of waiting for radiotherapy for grade III/IV gliomas. AB - AIM: To determine the effect of waiting time for radiotherapy on the overall survival of patients with high-grade gliomas. METHODS: We examined records of patients with grade III/IV gliomas who were referred to radiotherapy after surgery or biopsy - ECOG <3, any age, radical intent or palliative intent with dose >50 Gy, no interstitial or radiosurgery boost. Waiting time was defined in two ways, time from biopsy to radiotherapy and time from presentation to radiotherapy department to start of radiotherapy. RESULTS: There were 182 patients in the study having a median survival of 8.5 months, with a median follow up of 10.5 months. The group comprised of 63 (35%) grade III and 119 (65%) grade IV gliomas. Median times and ranges from biopsy and presentation to treatment were 26 days (4-78 days) and 15 days (1-62 days), respectively. The median dose was 60 Gy in a median of 30 fractions over a median of 46 days. Tumour progression before and during radiotherapy occurred in seven patients (4%) and 19 patients (11%), respectively. One hundred and seventy-nine patients died of disease. The seven patients whose tumour progressed before radiotherapy were excluded from the analysis of prognostic variables. In a multivariate analysis the variables that were significantly associated with worse survival were older age, reduced dose and prolonged waiting time from presentation. The risk of death increased by 2% for each day of waiting for radiotherapy. CONCLUSION: The study showed longer waiting time from presentation at radiotherapy department to treatment to be a significant predictor of overall survival for patients with high-grade glioma. PMID- 11054517 TI - Changes in the practice of adjuvant radiotherapy in resectable rectal cancer within a French well-defined population. AB - BACKGROUND AND PURPOSE: To assess the use of adjuvant radiotherapy in treating rectal cancers at a population level. MATERIALS AND METHODS: From 1976 to 1996, the influence of the period of diagnosis, sex, age, type of surgical resection, place of surgical resection on the use of radiotherapy was studied. A non conditional logistic regression was performed to obtain the odds radio for each studied period adjusted for the other variables. RESULTS: The use of adjuvant radiotherapy increased over time from 14.3% in 1976-1978 to 61.7% in 1994-1996 (odds ratio (OR): 28.0 for the 1994-1996 period compared with 1976-1978). It was also influenced by age (OR: 0.26 for patients >74 years compared with those <65 years), type of resection (OR: 3.42 for abdominoperineal resection compared with anterior resection) and place of surgery (OR: 0.39 for non-university hospitals compared with university hospitals). The nature of adjuvant radiotherapy altered over time: most adjuvant radiotherapy being done postoperatively before 1988, then preoperatively subsequently. CONCLUSIONS: Substantial changes have occurred in both the degree of use of adjuvant radiotherapy and in its timing. Some progress is still possible, in particular in older patients and in patients treated in non-university hospitals. PMID- 11054518 TI - Four radiation hypersensitivity cases and their implications for clinical radiotherapy. AB - BACKGROUND AND PURPOSE: Over a 20 year period, four out of 2000 paediatric radiotherapy patients, treated at St. Bartholomew's Hospital (three with lymphoma, one with angiosarcoma), have revealed extreme/fatal clinical hypersensitivity in normal tissues. PATIENTS AND METHODS: Cellular hypersensitivity was confirmed in vitro and attributed to the ataxia telangiectasia (A-T) gene in cases I and II, a newly described defect in the DNA ligase 4 gene in case III, and a novel and as yet incompletely defined, molecular defect in case IV who presented with xeroderma pigmentosum (XP). RESULTS: The severe clinical hypersensitivity preceded the cellular and molecular analysis, but did not manifest as a clinically exaggerated normal tissue reaction until 3+ weeks after the start of a conventionally fractionated course of radiotherapy, by which time the latent damage had been inflicted. There were no clinical stigmata to alert the clinician to a predisposing syndrome in two patients (cases I and II). We point out that approximately 20% of A-T patients are classified as variants with delayed expression of clinical symptoms, and case II falls into this category. CONCLUSIONS: As lymphoma (incidence, one in 100000 children) constituted the majority of the diagnoses, questions arise as to: (1), the probability of other centres having experienced and being presented in the future with similar problems (particularly bearing in mind that other oncologically predisposing radiosensitivity syndromes have not been not represented in our experience); and (2), the appropriateness, efficiency and applicability of predictive assays. Unambiguous cellular radiosensitivity would have been apparent from clonal assays on fibroblast cultures from all four cases prior to treatment, but such assays take 4-6 weeks to produce results. While estimates of chromosome damage or clonal assays on pre-treatment blood derived cells would be faster, there is a health economics issue as to the general applicability of such 'screening' assays. PMID- 11054519 TI - Improved local control for early T-stage nasopharyngeal carcinoma--a tale of two hospitals. AB - PURPOSE: To study the efficacy of intracavitary brachytherapy (ICT) in early T stage nasopharyngeal carcinoma (NPC). METHODS AND MATERIALS: All early T-stage (T1 and T2 nasal cavity tumour) NPC treated with a curative intent up to 1996 were analyzed (n=743), 163 from the Prince of Wales Hospital (PWH) and 25 from Tuen Mun Hospital (TMH) were given ICT after radical external radiotherapy (ERT; group A). They were compared with 555 patients treated with ERT alone (group B). The radiotherapy techniques were identical between the two hospitals. The ERT delivered the tumoricidal dose (uncorrected biological equivalent dose (BED)-10, > or = 75 Gy) to the primary tumour, and this did not differ in technique or dosage between the two groups. The ICT delivered a dose of 18-24 Gy in three fractions over 15 days to a point 1 cm perpendicular to the midpoint of the plane of the sources. RESULTS: The local failure was significantly less (crude rates, 6.9 vs. 13.0%; 5-year actuarial rates, 5.8 vs. 11.7%) and the disease-specific mortality was significantly lower (crude rates, 13.8 vs. 18.9%; 5-year actuarial rates, 12.2 vs. 15.2%) in group A compared with group B. ICT was the only significant independent prognostic factor predictive of fewer local failures. When ICT was excluded from the Cox regression model, the total physical dose or the total BED-10 uncorrected for tumour repopulation became significant in predicting the ultimate local failure rate. The two groups were comparable in the rate of the chronic radiation complications. A significant dose-tumour-control relationship existed, plotting the local failure as a function of the total physical dose or the total BED. CONCLUSIONS: Supplementing ERT, which delivered the tumoricidal dose (uncorrected BED-10, > or = 75 Gy), with ICT significantly enhanced ultimate local control in early T-stage (T1/T2 nasal infiltration) NPC. A significant dose-tumour-control relationship exists above the conventional tumoricidal dose level. PMID- 11054520 TI - Carcinoma of the maxillary antrum: a retrospective analysis of 110 cases. AB - BACKGROUND AND PURPOSE: Cancer of the maxillary antrum is a rare disease with a variety of treatment options. The present study was undertaken to review the outcome of patients with carcinoma of the maxillary antrum managed at a single institution. MATERIALS AND METHODS: A retrospective analysis of 110 cases of carcinoma of the maxillary antrum managed with curative intent during the time period 1976-1993 was performed. There were 33 females and 77 males; the median age was 64 years (range 38-89). The median follow-up time was 4 years (range from 2 months to 17 years). The majority of patients presented with locally advanced disease (78 T4 tumours); nodal involvement was observed in 17/110 cases. Histologic subtypes included in the analysis were limited to squamous cell carcinoma (95 cases) and undifferentiated carcinoma (15 cases). Patients were managed with either primary radiation therapy with surgery reserved for salvage (83/110) or with a planned combined approach with surgery and either pre or postoperative radiation (27/110). RESULTS: The actuarial 5-year cause-specific survival rate was 43%. The 5-year local control rate was 42%. Of 63 patients with local failure, 25 underwent salvage surgery with a subsequent 5-year cause specific survival of 31%. Multiple regression analysis of patient, disease and treatment related variables identified local disease extent and nodal disease at presentation as the only variables independently associated with cause-specific survival. CONCLUSIONS: This analysis indicates that survival from carcinoma of the maxillary antrum is poor with outcome strongly related to local disease extent. The best treatment strategy for this disease remains undefined. Salvage surgery can result in prolonged survival in selected patients experiencing local failure. PMID- 11054521 TI - Dermatofibrosarcoma protuberans: treatment results of 35 cases. AB - BACKGROUND AND PURPOSE: This study evaluates the treatment results of patients with dermatofibrosarcoma protuberans. PATIENTS AND METHODS: Between August 1987 and July 1998, 35 consecutive patients with pathologically proved dermatofibrosarcoma protuberans received surgery with or without radiation therapy. Their treatment results were analyzed retrospectively. RESULTS: The patient ages ranged from 5 to 67 years (median 37 years). There were 24 males and 11 females. The anatomic sites of tumor were: trunk in 21, extremity in eight, and head and neck region in six. The maximal dimension of tumor ranged from 1.5 to 25 cm. Surgery techniques included local excision and wide excision with or without graft or flap. Adjuvant radiation therapy was given to 11 patients, with a dose ranging from 46 to 68 Gy (one pre-operative, ten post-operative). At a median follow-up of 50 months (range 11-131 months), there were 11 patients (nine patients without radiation therapy) who developed local failure. Salvage therapy (excision with or without radiation therapy) was given to all of them, and ten achieved disease control. Some patients had treatment-related moderate cosmetic or functional problems. CONCLUSIONS: Dermatofibrosarcoma protuberans is a malignancy of a high cure rate, and adjuvant radiation therapy can reliably decrease the local recurrence rate and prevent mutilation and functional deficit caused by repeated surgery. PMID- 11054522 TI - A quantitative treatment planning study evaluating the potential of dose escalation in conformal radiotherapy of the oesophagus. AB - BACKGROUND AND PURPOSE: This study aims to evaluate the reduction in radiation dose to normal thoracic structures through the use of conformal radiotherapy techniques in the treatment of oesophageal cancer, and to quantify the resultant potential for dose escalation. MATERIALS AND METHODS: Three different CT-derived treatment plans were created and compared for each of ten patients. A two-phase treatment with conventional straight-edged fields and standard blocks (CV2), a two-phase conformal plan (CF2), and a three-phase conformal plan where the third phase was delivered to the gross tumour only (CF3), were considered for each patient. Escalated dose levels were determined for techniques CF2 and CF3, which by virtue of the conformal field shaping, did not increase the mean lung dose. The resulting increase in tumour control probability (TCP) was estimated. RESULTS: A two-phase conformal technique (CF2) reduced the volume of lung irradiated to 18 Gy from 19.7+/-11.8 (1 SD) to 17.1+/-12.3% (P=0.004), and reduced the normal tissue complication probability (NTCP) from 2.4+/-4.0 to 0.7+/ 1.6% (P=0.02) for a standard prescribed dose of 55 Gy. Consequently, technique CF2 permitted a target dose of 59.1+/-3.2 Gy without increasing the mean lung dose. Technique CF3 facilitated a prescribed dose of 60.7+/-4.3 Gy to the target, the additional 5 Gy increasing the TCP from 53. 1+/-5.5 to 68.9+/-4.1%. When the spinal cord tolerance was raised from 45 to 48 Gy, technique CF3 allowed 63.6+/ 4.l Gy to be delivered to the target, thereby increasing the TCP to 78.1+/-3.2%. CONCLUSIONS: Conformal radiotherapy techniques offer the potential for a 5-10 Gy escalation in dose delivered to the oesophagus, without increasing the mean lung dose. This is expected to increase local tumour control by 15-25%. PMID- 11054523 TI - Computed tomography/magnetic resonance based volume changes of the primary tumour in patients with prostate cancer with or without androgen deprivation. AB - BACKGROUND AND PURPOSE: To evaluate changes of the volume of the cancerous prostatic gland during androgen deprivation (AD) started immediately after diagnosis (IAD). Hypothetically, these data would assist the radiotherapist to determine the appropriate duration of pre-radiotherapy downsizing neoadjuvant luteinizing hormone releasing hormone (LHRH) treatment. A second aim was to assess any increase of the prostatic volume during the 1st year of diagnosis in patients who were allocated to a deferred treatment policy (DAD). METHODS AND MATERIALS Thirteen patients in the IAD cohort and 13 patients in the DAD group, all with T1-3pN1-2M0 prostate cancer, had regular computed tomography/magnetic resonance (CT/MR) examinations during the 1st year after randomization within the EORTC-GU trial 30846. Pre-treatment prostate specific antigen (PSA) values were available in only 12 patients. RESULTS: In the IAD group the prostate gland decreased with significant difference as compared with the DAD patients (P=0.033). As compared with the pre-treatment situation the prostate gland in the IAD group was reduced in size by 18, 35, and 46% at 1, 6, and 12 months, respectively. In four of six evaluable IAD patients the prostatic volume continued to shrink after achievement of the nadir PSA level (at 3 months). In three of the 13 DAD patients the prostate volume increased by >25% during the 1st 3 months after randomization. CONCLUSION: If neoadjuvant androgen deprivation is applied before local treatment to downsize the volume of the cancerous prostate gland, our limited data suggest that such treatment should last at least 6 months in order to achieve a maximal effect in the majority of patients. In about 1/4 of untreated patients an increase in the prostate volume by >25% may occur within 3 months of diagnosis. If no AD is given, radiotherapy should start within this period. PMID- 11054524 TI - Improving the preservation of erectile function after external beam radiation therapy for prostate cancer. AB - BACKGROUND AND PURPOSE: To investigate whether the type of collimation technique, target dose and treated volume influence the prevalence of intact erectile function after external beam radiation therapy for localized prostate cancer. PATIENTS AND METHODS: A prospective study was conducted to assess erection stiffness before treatment and after follow-ups of 9-18 months and 4-5.5 years. Information was collected using the Radiumhemmet Scale of Sexual Function. RESULTS: Thirty-one men were 'potent' before the radiation. Fourteen of them were treated with a conventional collimator and 17 were given three-dimensional conformal therapy with the aid of a multileaf collimator. Preserved erectile function at 9-18 months was found in 17 of the 31 men (55%) and at the 4-5-year follow-up in five of 22 (23%). Preservation of potency was related to the treatment procedure but not to the treatment volume. CONCLUSIONS: Conformal therapy may increase the percentage of men preserving erectile function during radiotherapy for localized prostate cancer; it is possible that the differences to conventional therapy do not depend on treated volume. PMID- 11054525 TI - Solitary brain metastases treated with the Leksell gamma knife: prognostic factors for patients. AB - PURPOSE: To analyze treatment results, complications, prognostic factors and their statistical significance in surviving patients treated with the Leksell gamma knife (LGK) for solitary brain metastases. MATERIALS AND METHODS: Between 1992-1998, 237 patients were treated with solitary brain metastasis (SBM). The histological subtypes were as follows: 101 patients (42.6%) non small-cell lung cancer, 42 (17.7%) renal cell carcinoma, 36 (15.2%) breast carcinoma, 30 (12.7%) colorectal carcinomas and 28 (11.8%) melanoma. RESULTS: A complete or partial regression was observed in 193 (81.4%) patients, cessation of growth activity in 32 (13.5%) and local progression in 12 (5.1%). Local recurrence was observed in nine (4.7%) of 193 patients with complete or partial regression. Acute toxicity appeared in 24 (10%) patients (score 3.4), late complications (score 3.4) were observed in 13 patients (5.5%). Out of 237 patients, 182 patients died with a median survival of 6 months and 55 patients are still alive with a median survival 12 months and with a minimal follow up period of 10 months. The significant prognostic factors for longer survival in these series of patients were: Karnofsky performance status (70% or more), the extent of extra-cerebral disease (no extra-cerebral disease), pretreatment neurological symptoms neurological functional class (NFC 1), histology (renal cell and breast carcinomas) and the dose to the planning target volume (20 Gy and higher). CONCLUSIONS: Radiosurgery provides an effective local control for 90% of treated patients with low morbidity. Several significant prognostic factors were detected for patients' survival. These factors can help to predict better patients' survival after the LGK treatment. PMID- 11054526 TI - Fractionated stereotactic conformal radiation therapy of brain stem gliomas: outcome and prognostic factors. AB - BACKGROUND AND PURPOSE: Evaluation of outcome and prognostic factors in patients with brain stem glioma (BSG) following fractionated stereotactic radiotherapy (FSRT). MATERIALS AND METHODS: Between 1990 and 1997, we treated 41 patients with FSRT in a phase I/II trial. Median age was 24 years. Out of 36 patients with histologically proven glioma, ten had a partial tumour resection. Histology revealed low grade gliomas in 30 patients and anaplastic gliomas in six patients. A mean total dose of 54 Gy was given in daily fractions of 1.8 Gy. Median follow up was 12 months. RESULTS: Three patients died during FSRT. Neurological improvement was achieved in 19/38 patients. Reduction of tumour size was reported in 12/38, in 16 patients the lesion was unchanged, ten showed progression. Median time to progression was 23 months, median overall survival 40 months with an actuarial survival of 83% at 1 year, 55% at 3 years and 33% at 5 years. In 20 of 22 patients with recurrence progression was inside the target volume. Significant prognostic factors for survival were clinical and radiological response 6 weeks after FSRT. Treatment toxicity was mild. Ototoxicity occurred in one patient. CONCLUSIONS: FSRT is a feasible treatment modality for BSG with tolerable toxicity. The risk of marginal failure is low. PMID- 11054527 TI - Stereotactic radiotherapy of extracranial targets: CT-simulation and accuracy of treatment in the stereotactic body frame. AB - BACKGROUND AND PURPOSE: Evaluation of set-up accuracy and analysis of target reproducibility in the stereotactic body frame (SBF), designed by Blomgren and Lax from Karolinska Hospital, Stockholm. Different types of targets were analyzed for the risk of target deviation. The correlation of target deviation to bony structures was analyzed to evaluate the value of bones as reference structures for isocenter verification. MATERIALS AND METHODS: Thirty patients with 32 targets were treated in the SBF for primary or metastatic peripheral lung cancer, liver metastases, abdominal and pelvic tumor recurrences or bone metastases. Set up accuracy and target mobility were evaluated by CT-simulation and port films. The contours of the target at isocenter level, bony structures and body outline were compared by matching the CT-slices for treatment planning and simulation using the stereotactic coordinates of the SBF as external reference system. The matching procedure was performed by using a 3D treatment planning program. RESULTS: Set-up accuracy represented by bony structures revealed standard deviations (SD) of 3.5 mm in longitudinal, 2.2 mm in anterior-posterior and 3.9 mm in lateral directions. Target reproducibility showed a SD of 4.4 mm in longitudinal, 3.4 mm ap and 3.3 mm in lateral direction prior to correction. Correlation of target deviation to bones ranged from 33% (soft tissue targets) to 100% (bones). CONCLUSION: A security margin of 5 mm for PTV definition is sufficient, if CT simulation is performed prior to each treatment to correct larger target deviations or set-up errors. Isocenter verification relative to bony structures is only safe for bony targets but not for soft tissue targets. PMID- 11054528 TI - Reproducibility of geometric distortion in magnetic resonance imaging based on phantom studies. AB - BACKGROUND AND PURPOSE: Image distortion is one of the major drawbacks of magnetic resonance (MR) imaging for use in radiotherapy treatment planning (RTTP). In this study, the reproducibility of MR imaging distortion was evaluated by repeated phantom measurements. MATERIALS AND METHODS: A grid-pattern acrylic phantom was scanned with a 0.2-Tesla permanent magnetic unit. We repeated a series of scans three times to evaluate the reproducibility of the distortion. In each series, co-ordinates at 432 intersections of the grid were measured for both T1- and T2-weighted spin-echo (SE) pulse sequences. Positional displacements and their variations at the intersections were calculated. RESULTS: Averages of the displacements were distributed between 1.58 and 1.74 mm, and maximum values (MAX) between 12.6 and 15.0 mm. Within 120 mm of the image center, the average values ranged from 0.73 to 0.80 mm, and from 3.4 to 5.0 mm for MAX. The absolute values of the positional variations among three series were distributed between 0.41 and 0.88 mm for average values, and between 1.4 and 4.5 mm for MAX. CONCLUSIONS: The positional variations were mostly within 3 pixels, and most of the positional displacements within the radius of 120 mm of the image center were 2 mm or less. Therefore, it will be possible to use this MR system in RTTP under limited situations, although careful applications are required for RTTP of the body. The development of a computer program to correct image distortion is expected. PMID- 11054529 TI - Cloning and characterization of the human ameloblastin gene. AB - We isolated the full-length human ameloblastin (AMBN) cDNA clone using reverse transcription-polymerase chain reaction (RT-PCR) methods. Sequence analysis of the AMBN cDNA revealed an open reading frame of 1341bp encoding a 447-amino-acid protein. Comparison with pig, cattle, rat, and mouse AMBN sequences showed a high amino acid sequence similarity and led to the identification of a novel 78bp (26 amino acids) insert resulting from internal sequence duplication. By DNA analysis of a human genomic clones, the AMBN gene was shown to consist of 13 exons and a novel 78bp segment, which proved to comprise two small exons. Human ameloblastomas express AMBN transcripts that contain some mutations. PMID- 11054530 TI - The gas 5 gene shows four alternative splicing patterns without coding for a protein. AB - The murine gas5 gene was originally isolated based on its preferential expression in the growth arrest phase of the cell cycle. This gene contains 12 exons from which two alternatively spliced transcripts have been initially identified. More recently, it has been reported that both human and murine gas5 genes contain in their introns sequences homologous to the small nucleolar RNAs involved in the processing of ribosomal RNA. Here we report on the identification and analysis of the expression pattern of two novel alternatively spliced mouse gas5 mRNAs which contain no obvious open reading frame (ORF). Using antibodies generated against the putative amino acid sequence deduced from the gas5 cDNAs, we were not able to detect any Gas5 protein in cultured cells or murine tissues extracts. Even more definitive evidence that the gas5 gene may not encode a protein was obtained by cloning and sequencing the rat gas5 gene which revealed that the putative ORF is interrupted by a stop codon after the first 13 amino acids. PMID- 11054531 TI - Kdap, a novel gene associated with the stratification of the epithelium. AB - The skin develops and differentiates during embryogenesis, which is concertedly regulated by a variety of genes. The present study isolated from the rat embryonic skin a novel differentiation-associated gene named Kdap (keratinocyte differentiation-associated protein) by suppression subtractive hybridization between the skin of 14day postcoitus (dpc) embryo (the prehair-germ stage) and that of 17dpc embryo (the hair-germ stage). Its mRNA contained four spliced forms in these tissues. The gene encoded a protein of total 98 amino acids with a calculated molecular mass of 11kDa and an isoelectric point of 6.1 as an unspliced form. The two splicing zones were well conserved among rat, mouse, and human. This protein had a high hydrophobic N-terminal region, a possible signal sequence, and contained two putative N-myristoylation sites and two casein kinase II phosphorylation sites. In situ hybridization experiments detected Kdap transcripts exclusively in the suprabasal cell layers of the embryonic epidermis. Intense expression was also seen in suprabasal cells in regions of infundibulum of the hair follicle. These results indicated that Kdap provides a new insight into the mechanism of differentiation and the maintenance of stratified epithelia. PMID- 11054532 TI - Formation of regulator/target gene relationships during evolution. AB - The Foxn1-like forkhead/winged-helix transcription factor genes have been maintained in single copy throughout chordate evolution. Among other functions, Foxn1 (formerly known as Whn) regulates the expression of hair keratin genes in the hair follicle, which represents an evolutionarily novel organ characteristic of mammals. We show here that fish and mouse Foxn1-like genes are functionally equivalent in hair keratin gene activation, suggesting the absence of functionally relevant changes over the course of several hundred million years of vertebrate evolution. In contrast, the Foxn1-like gene from the cephalochordate Branchiostoma lanceolatum is inactive in this assay because of changes in the region located N-terminal to DNA binding and transcriptional activation domains of the protein. Our results indicate that functionally relevant changes in cis regulatory regions are not necessarily accompanied by corresponding changes in transcription factor proteins in the formation of evolutionarily novel regulator/target gene relationships. PMID- 11054533 TI - dfh is a Drosophila homolog of the Friedreich's ataxia disease gene. AB - A putative Drosophila homolog of the Friedreich's ataxia disease gene (FRDA) has been cloned and characterized; it has been named Drosophila frataxin homolog (dfh). It is located at 8C/D position on X chromosome and is spread over 1kb, a much smaller genomic region than the human gene. Its genomic organization is simple, with a single intron dividing the coding region into two exons. The predicted encoded product has 190 amino acids, being considered a frataxin-like protein on the basis of the sequence and secondary structure conservation when compared with human frataxin and related proteins from other eukaryotes. The closest match between the Drosophila and the human proteins involved a stretch of 38 amino acids at C-terminus, encoded by dfh exon 2, and exons 4 and 5a of the FRDA gene, respectively. This highly conserved region is very likely to form a functional domain with a beta sheet structure flanked by alpha-helices where the sequence is less conserved. A signal peptide for mitochondrial import has also been predicted in the Drosophila frataxin-like protein, suggesting its mitochondrial localization, as occurs for human frataxin and other frataxin-like proteins described in eukaryotes. The Drosophila gene is expressed throughout the development of this organism, with a peak of expression in 6-12h embryos, and showing a spatial ubiquitous pattern from 4h embryos to the last embryonic stage examined. The isolation of dfh will soon make available specific dfh mutants that help in understanding the pathogenesis of FRDA. PMID- 11054534 TI - Molecular structure and evolution of DNA sequences located at the alpha satellite boundary of chromosome 20. AB - We have isolated and characterised one PAC clone (dJ233C1) containing a linkage between alphoid and non-alphoid DNA. The non-alphoid DNA was found to map at the pericentromeric region of chromosome 20, both on p and q sides, and to contain homologies with one contig (ctg176, Sanger Centre), also located in the same chromosome region. At variance with the chromosome specificity shown by the majority of non-alphoid DNA, a subset of alphoid repeats derived from the PAC yielded FISH hybridisation signals located at the centromeric region of several human chromosomes, belonging to three different suprachromosomal families. The evolutionary conservation of this boundary region was investigated by comparative FISH experiments on chromosomes from great apes. The non-alphoid DNA was found to have undergone events of expansion and transposition to different pericentromeric regions of great apes chromosomes. Alphoid sequences revealed a very wide distribution of FISH signals in the great apes. The pattern was substantially discordant with the data available in the literature, which is essentially derived from the central alphoid subset. These results add further support to the emerging opinion that the pericentromeric regions are high plastics, and that the alpha satellite junctions do not share the evolutionary history with the main subsets. PMID- 11054535 TI - Cloning, expression analysis, and chromosomal mapping of GTPBP2, a novel member of the G protein family. AB - We have identified a novel gene encoding a protein bearing GTP-binding motifs, the characteristics of GTP-binding proteins (G proteins). The deduced amino acid sequence exhibited the highest overall homology with GTPBP1 and its mouse orthologue GP-1. Hence, we named the gene GTPBP2. The mouse orthologue of this gene, Gtpbp2, showed 98% identity with GTPBP2 over the entire protein (the HGMW approved nomenclature symbol is GTPBP2 and mouse orthologue is Gtpbp2). A phylogenetic analysis showed GTPBP2 and homologous G proteins (GTPBP1, AGP-1, and CGP-1) did not belong to major G protein families. They formed a distinct branch in the phylogenetic tree, suggesting that they constitute a novel G protein family. A 2. 9kb mRNA was predominantly detected in the testis along with various other organs. In situ hybridization analysis revealed that Gtpbp2 was predominantly expressed in spermatocytes and round-spermatids in the testis. These novel genes were localized to human chromosome 6p21.1-2 and mouse chromosome 17qC-D. PMID- 11054536 TI - In vivo expression and genomic organization of the mouse cyclin I gene (Ccni). AB - Cyclins control cell-cycle progression by regulating the activity of cyclin dependent kinases. Cyclin I was recently added to the cyclin family of proteins because of the presence of a cyclin box motif in the deduced amino-acid sequence. Cyclin I may share functional roles with cyclin G1 and G2 because of the high structural similarity between their deduced amino-acid sequences. However, the biological and functional roles of this subclass of cyclins remain obscure. The mouse cyclin G1 and G2 genes have previously been cloned and characterized. In this report, we describe the cloning of the mouse homolog of cyclin I. The cyclin I cDNA sequence was used to determine the genomic organization of the mouse cyclin I gene which co-localizes with cyclin G2 to chromosome 5E3.3-F1.3. Cyclin I was transcribed from seven exons distributed over more than 19kb of genomic sequence. The expression of cyclin I was determined in various tissues, but no clear correlation with the proliferative state was found. Furthermore, in contrast to cyclin G1, cyclin I expression was stable during cell-cycle progression after partial hepatectomy in both the absence and presence of DNA damage. Transient expression of cyclin I-green fluorescent protein (GFP) fusion proteins in cell lines showed that cyclin I was distributed throughout the cell in contrast with the mainly cytoplasmic localization of cyclin G2 and nuclear localization of cyclin G1. Our results indicate that despite the close structural similarity between cyclin G1, G2 and I, these three proteins are likely to have distinct biological roles. PMID- 11054537 TI - Analysis of the murine phosphoinositide 3-kinase gamma gene. AB - Phosphoinositide 3-kinase gamma is preferentially expressed in leukocytes. PI3Kgamma is activated by betagamma subunits of heterotrimeric G-proteins, which thus link seven transmembrane helix receptor activation to phosphatidylinositol (3,4,5)-trisphosphate production. Here we describe the molecular cloning of the murine PI3Kgamma cDNA, the PI3Kgamma gene structure, its chromosomal assignment and the analysis of promoter activity. The mouse cDNA shares 86% identity to its pig and human orthologues at the nucleotide level. The MmPI3Kgamma gene spans approximately 30kb and comprises 11 exons. RACE-PCR indicated the presence of multiple start sites generating 5' UTRs with different lengths, the longest being 874bp. The putative promoter region contains no TATA box but several putative binding sites for hematopoietic specific transcription factors. A 1200bp long sequence upstream the first transcription start site was found to possess tissue specific promoter activity. Deletion constructs revealed two contiguous regions, with activator function, ranging from positions -139 to -557, and with inhibitory function, ranging from positions -557 to -892. FISH analysis revealed that the MmPI3Kgamma is located on chromosome 12 band B and that the human orthologue is positioned on chromosome 7q22.2-22.3. In spite of some differences in the ATP binding site, recombinant murine PI3Kgamma protein is equally sensitive to wortmannin as its human counterpart. This suggests that mouse models will provide reliable results in the assessments of novel PI3Kgamma inhibitors. PMID- 11054538 TI - A cytochrome P4501B gene from a fish, Pleuronectes platessa. AB - Tetrapod cytochrome P4501 family (CYP1A1, CYP1A2 and CYP1B1) enzymes are most active in hydroxylating a variety of environmental contaminants including polyaromatic hydrocarbons (PAH), planar polychlorinated biphenyls and arylamines and thus play a pivotal role in the toxicology of these compounds. Mammalian CYP1A1 and CYP1A2 genes appear to have diverged after the evolutionary emergence of mammals, whereas fish species apparently possess only one CYP1A family gene, and fish CYP1A enzymes exhibit properties of both of the mammalian isoforms. We have isolated a further CYP1 family gene from a marine flatfish (plaice; Pleuronectes platessa), which, on the basis of exon organisation and sequence similarity, can be assigned as a piscine CYP1B. Its deduced amino acid sequence shows the closest (54%) identity to mammalian CYP1B1 proteins and, on the basis of molecular modeling studies, shows a high degree of positional and structural conservation of the substrate contacting amino acid residues in its putative active site when compared to other CYP1 enzymes. Phylogenetic analysis of fish and mammalian CYP1 family sequences indicates that the plaice CYP1B and mammalian CYP1B1 genes share a common ancestry. Plaice CYP1B has a more restricted tissue expression profile than the previously isolated plaice CYP1A, only being detectable, by Northern blotting, in gill tissue. In contrast to CYP1A, which shows extensive PAH-dependent induction in a variety of tissues, plaice CYP1B appears unresponsive to treatment with a prototypical PAH-type inducer, beta naphthoflavone (BNF). PMID- 11054539 TI - Molecular cloning and expression during development of the Drosophila gene for the catalytic subunit of DNA polymerase epsilon. AB - We have cloned the genomic DNA and cDNA of Drosophila DNA polymerase epsilon (pol epsilon) catalytic subunit (GenBank No. AB035512). The gene is separated into four exons by three short introns, and the open reading frame consists of 6660 base pairs (bp) capable of encoding a polypeptide of 2220 amino acid residues. The calculated molecular mass is 255018, similar to that of mammalian and yeast homologues. The deduced amino acid sequence of the pol-epsilon catalytic subunit shares approximately 41% identity with human and mouse homologues as well as significant homology those of C. elegans, S. cerevisiae and S. pombe. Similar to the pol-epsilon catalytic subunits from other species, the pol-epsilon catalytic subunit contains domains for DNA polymerization and 3'-5' exonuclease in the N terminal region, and two potential zinc-finger domains in the C-terminal regions. Interestingly, a 38 amino acid sequence in the C-terminal region from amino acid positions 1823 to 1861 is similar to the site for Mycoplasma ATP binding and/or ATPase domain (GenBank No. P47365). Northern hybridization analysis indicated that the gene is expressed at the highest levels in unfertilized eggs, followed by zero to 4h embryos and adult females, and then embryos at other embryonic stages, instar larva stages and adult males. Low levels of the mRNA were also detected at the pupa stage. This pattern of expression is similar to those of DNA replication-related enzymes such as DNA polymerase alpha and delta except for the high level of expression in adult males. PMID- 11054540 TI - The murine ortholog of matrix metalloproteinase 19: its cloning, gene organization, and expression. AB - We have isolated a murine cDNA orthologous to the human matrix metalloproteinase 19 (hMMP-19). The murine MMP-19 cDNA was amplified by RT-PCR using specific primers whose DNA sequences were derived from both murine MMP-19 genomic DNA and partial cDNA sequences. The murine MMP-19 (mMMP-19) is 79% identical to the human ortholog and encodes a protein of 527 amino acids with a deduced molecular mass of 59.1kDa. Analyzing the exon/intron junctions we revealed that the murine MMP 19 gene consists of nine exons and eight introns, and thus differs from the gene organization of other matrix metalloproteinases. Furthermore, a 587bp fragment of the mMMP-19 promoter containing a TATA box and an AP-1 binding motif was cloned, and 3.3kb transcripts of the MMP-19 gene were identified in liver, kidney, spleen, and colon. Finally, immunostaining of murine heart cryosections showed that mMMP-19, like its human counterpart, is expressed in the arterial tunica media of large blood vessels. By cloning mMMP-19 and unraveling its genomic structure, we have obtained valuable information for further study of the function of this MMP in vivo. PMID- 11054541 TI - Cloning and characterization of AWP1, a novel protein that associates with serine/threonine kinase PRK1 in vivo. AB - We describe the cloning and expression of cDNAs encoding a novel human protein of 208 amino acid residues with a predicted molecular mass of 22.6kDa and its mouse homologue. We name this protein as AWP1 (associated with PRK1). AWP1 is a ubiquitously expressed protein, and the Awp1 gene is switched on during early human and mouse development. When expressed in COS-1 cells, the Myc-tagged AWP1 has an apparent molecular mass higher than that deduced from its amino acid sequence. AWP1 possesses a conserved zf-A20 zinc finger domain at its N-terminal and a zf-AN1 zinc finger domain at its C-terminal. Co-immunoprecipitation experiments revealed that mouse AWP1 specifically interacts with a rat serine/threonine protein kinase PRK1 in vivo. Hence, AWP1 may play a regulatory role in mammalian signal transduction pathways. PMID- 11054542 TI - Regulation of the promoters for the human bone morphogenetic protein 2 and 4 genes. AB - The bone morphogenetic proteins 2 and 4 are known to be important in bone formation and are expressed in both the developing and adult mammalian bone. Understanding the regulation of these genes in osteoblasts may yield methods by which we can control expression to induce bone formation. We have isolated and characterized the human BMP-2 and BMP-4 promoters and report substantially more upstream sequence information than that which has been published. Human osteoblasts were found to have a single transcript initiation site that is conserved across species, rather than multiple start sites, as has previously been reported (Feng, J.Q., Harris, M.A., Ghosh-Choudhury, N., Feng, M., Mundy, G.R., Harris, S.E., 1994. Structure and sequence of mouse morphogenetic protein-2 gene (BMP-2): comparison of the structures and promoter regions of BMP-2 and BMP 4 genes. Biochim. Biophys. Acta 1218, 221-224; Heller, L.C., Li, Y., Abrams, K.L., Rogers, M.B., 1999. Transcriptional regulation of the Bmp2 gene. J. Biol. Chem. 274, 1394-1400; Sugiura, T., 1999. Cloning and functional characterization of the 5'-flanking region of the human bone morphogenetic protein-2 gene. Biochem. J. 338, 433-440). A series of promoter deletions for both human BMP-2 and BMP-4 fused to the luciferase reporter gene were analyzed thoroughly in human and murine osteoblastic cell lines. Several compounds and growth factors that stimulate general or osteogenic pathways were used to treat cells transfected with the promoter constructs. Retinoic acid compounds and the phorbol ester, PMA were found to stimulate BMP-2 and, to a lesser degree, BMP-4. The combination of all trans-RA and PMA caused a synergistic increase in BMP-2 promoter activity and endogenous mRNA. The RA stimulation appears to be an indirect effect on the BMP-2 promoter, as the most highly conserved RRE in the BMP-2 promoter was unable to functionally bind or compete for protein binding. Potential binding sites in both promoters for the bone-specific transcription factor, Cbfa-1, were found to specifically bind Cbfa-1 protein in osteoblast nuclear extracts; however, deletion of these sites did not significantly affect transcriptional activity of the promoters in osteoblasts. These data thus present new sequence and regulatory information for the human BMP-2 and BMP-4 promoters and clarify the human BMP-2 gene transcriptional start site in osteoblasts. PMID- 11054543 TI - cDNA cloning, chromosomal localization, and expression analysis of human BEHAB/brevican, a brain specific proteoglycan regulated during cortical development and in glioma. AB - BEHAB (Brain Enriched HyAluronan Binding)/brevican, a brain-specific member of the lectican family of chondroitin sulfate proteoglycans (CSPGs), may play a role in both brain development and human glioma. BEHAB/brevican has been cloned from bovine, mouse and rat. Two isoforms have been reported: a full-length isoform that is secreted into the extracellular matrix (ECM) and a shorter isoform with a sequence that predicts a glycophosphatidylinositol (GPI) anchor. Here, we report the characterization of BEHAB/brevican isoforms in human brain. First, BEHAB/brevican maps to human chromosome 1q31. Second, we report the sequence of both isoforms of human BEHAB/brevican. The deduced protein sequence of full length, secreted human BEHAB/brevican is 89.7, 83.3 and 83.2% identical to bovine, mouse and rat homologues, respectively. Third, by RNase protection analysis (RPA) we show the developmental regulation of BEHAB/brevican isoforms in normal human cortex. The secreted isoform is highly expressed from birth through 8years of age and is downregulated by 20years of age to low levels that are maintained in the normal adult cortex. The GPI isoform is expressed at uniformly low levels throughout development. Fourth, we confirm and extend previous studies from our laboratory, here demonstrating the upregulation of BEHAB/brevican mRNA in human glioma quantitatively. RPA analysis shows that both isoforms are upregulated in glioma, showing an approximately sevenfold increase in expression over normal levels. In contrast to the developmental regulation of BEHAB/brevican, where only the secreted isoform is regulated, both isoforms are increased in parallel in human glioma. The distinct patterns of regulation of expression of the two isoforms suggest distinct mechanisms of regulation of BEHAB/brevican during development and in glioma. PMID- 11054544 TI - Characterization of fimN, a new Bordetella bronchiseptica major fimbrial subunit gene. AB - Fimbrial proteins play an important role in the binding of Bordetella bronchiseptica to mammalian cells, an event that is key to the pathogenesis of this organism. The fimbrial phenotype of B. bronchiseptica isolates is usually defined serologically by Fim2 and Fim3 antigens. In this study, a previously unidentified fimbrial gene, fimN, was cloned and sequenced. The identity of fimN is based on several observations. The predicted FimN protein has 59.4 and 52. 2% homology with B. bronchiseptica Fim2 and Fim3, respectively, and is similar in size to these fimbriae. fimN, expressed as a recombinant protein, is recognized by mAb prepared against Fim2 from Bordetella pertussis. The fimN promoter region contains a stretch of cytosine residues similar in length to those of other fimbrial genes expressed by Bordetella species. It also has an activator binding region, upstream from the C-stretch, that closely resembles a corresponding bvg regulated region in fim2, fim3, and fimX. The fimN gene was isolated from a cosmid prepared with B. bronchiseptica genomic DNA that restored normal properties of cellular adhesion to an adhesion deficient strain of B. bronchiseptica. As such, FimN may be a previously overlooked fimbrial antigen and may play an important role in the pathogenicity of B. bronchiseptica. PMID- 11054545 TI - Distribution of cognates of group II introns detected in mitochondrial cox1 genes of a diatom and a haptophyte. AB - We identified group IIA introns that contain an open reading frame (ORF) in the mitochondrial cytochrome oxidase subunit I (cox1) genes of yellow algae, a diatom Thalassiosira (Th.) nordenskioeldii CCMP 992 collected from the east coast of USA, and a haptophyte Pavlova (Pa.) lutheri CCMP 1325 collected from Finland. Cognate introns of CCMP 1325 were detected in all Pa. lutheri strains investigated, which were collected from various oceans. In contrast, the intron was absent from closely related species belonging to the same genus Pavlova. This was also the case for the group II intron detected in a diatom Th. nordenskioeldii CCMP 992. The group II intron of CCMP 992 was located at the corresponding site to the group IIA intron found in Pylaiella (synonym, Pilayella) littoralis. The deduced secondary structures of these introns, one of which is from a diatom and the other from a brown alga, were virtually identical. In contrast, the haptophyte group II intron was inserted at a novel locus, and shares no particularly high sequence homology with any intron known to date. The phylogenetic tree based on the intronic ORF domain was not congruent with that based on the cox1 exon. The most prominent property of the intronic ORF tree was that introns located at homologous sites made robust pair clades irrespective of the phylogenetic relationships of the organisms. This suggests that mitochondrial group II introns often invade intronless alleles across the species barrier with site specificity. Homology analysis of the haptophyte intronic ORF suggested that it comprises three domains: reverse transcriptase (RT), RNA maturase (Ma), and H N-H endonuclease. However, the intronic ORF of the diatom contains the Ma domain but is apparently missing the H-N-H domain, and its RT domain is most probably partly or completely lacking in function. PMID- 11054546 TI - Common phylogeny of catalase-peroxidases and ascorbate peroxidases. AB - Catalase-peroxidases belong to Class I of the plant, fungal, bacterial peroxidase superfamily, together with yeast cytochrome c peroxidase and ascorbate peroxidases. Obviously these bifunctional enzymes arose via gene duplication of an ancestral hydroperoxidase. A 230-residues long homologous region exists in all eukaryotic members of Class I, which is present twice in both prokaryotic and archaeal catalase-peroxidases. The overall structure of eukaryotic Class I peroxidases may be retained in both halves of catalase-peroxidases, with major insertions in several loops, some of which may participate in inter-domain or inter-subunit interactions. Interspecies distances in unrooted phylogenetic trees, analysis of sequence similarities in distinct structural regions, as well as hydrophobic cluster analysis (HCA) suggest that one single tandem duplication had already occurred in the common ancestor prior to the segregation of the archaeal and eubacterial lines. The C-terminal halves of extant catalase peroxidases clearly did not accumulate random changes, so prolonged periods of independent evolution of the duplicates can be ruled out. Fusion of both copies must have occurred still very early or even in the course of the duplication. We suggest that the sparse representatives of eukaryotic catalase-peroxidases go back to lateral gene transfer, and that, except for several fungi, only single copy hydroperoxidases occur in the eukaryotic lineage. The N-terminal halves of catalase-peroxidases, which reveal higher homology with the single-copy members of the superfamily, obviously are catalytically active, whereas the C-terminal halves of the bifunctional enzymes presumably control the access to the haem pocket and facilitate stable folding. The bifunctional nature of catalase peroxidases can be ascribed to several unique sequence peculiarities conserved among all N-terminal halves, which most likely will affect the properties of both haem ligands. PMID- 11054547 TI - Human serine racemase: moleular cloning, genomic organization and functional analysis. AB - High levels of D-serine are found in mammalian brain, where it is an endogenous agonist of the strichinine-insensitive site of N-methyl D-aspartate type of glutamate receptors. D-serine is enriched in protoplasmic astrocytes that occur in gray matter areas of the brain and was shown to be synthesized from L-serine. We now report cloning and expression of human serine racemase, an enzyme that catalyses the synthesis of D-serine from L-serine. The enzyme displays a high homology to the murine serine racemase. It contains a pyridoxal 5'-phosphate attachment sequence similar to bacterial biosynthetic threonine dehydratase. Northern-blot analysis show high levels of human serine racemase in areas known to contain large amounts of endogenous D-serine, such as hippocampus and corpus callosum. Robust synthesis of D-serine was detected in cells transfected with human serine racemase, demonstrating the conservation of D-amino acid metabolism in humans. Serine racemase may be a therapeutic target in psychiatric diseases as supplementation of D-serine greatly improves schizophrenia symptoms. We identify the human serine racemase genomic structure and show that the gene encompasses seven exons and localizes to chromosome 17q13.3. Identification of the intron exon boundaries of the human serine racemase gene will be useful to search for mutations in neuropsychiatric disorders. PMID- 11054548 TI - Murine Bv8 gene maps near a synteny breakpoint of mouse chromosome 6 and human 3p21. AB - The genomic structure of the murine Bv8 gene was determined in 129/SvJ mouse, and the chromosomal localization was identified. Bv8 has first been characterized from skin secretion of the yellow-bellied toad, Bombina variegata. When injected into rat brain, this polypetide causes hyperalgesia. The murine Bv8 gene was shown to consist of four exons and was localized on chromosome 6 between the microsatellite markers D6Mit66 and D6Mit36 near the gene mem1, whereas the human counterpart was assigned to the non-syntenic region 3p21.1. Furthermore, the primary Bv8 transcript appeared to be alternatively spliced. The first variant contained all four exons yielding a product with a stretch highly enriched in basic amino acids in its central part. This domain is absent in the peptides from frog as well as in a splice variant expressed in mouse testis. A third variant gives rise to a truncated polypeptide. PMID- 11054549 TI - Stable and unstable transgene integration sites in the human genome: extinction of the Green Fluorescent Protein transgene in K562 cells. AB - In gene transfer experiments including gene therapy studies, expression of the integrated transgenes in host cells often declines with time. The molecular basis of this phenomenon is not clearly understood. We have used the Green Fluorescent Protein (GFP) gene as both a selectable marker and a reporter to study long-term transgene integration and expression in K562 cells. Cells transfected with plasmids containing the GFP gene coupled to the HS2 or HS3 enhancer of the human beta-globin Locus Control Region (LCR) or the cytomegalovirus (CMV) enhancer were sorted by either fluorescence-activated-cell-sorting (FACS) alone or FACS combined with drug selection based on a co-integrated drug resistance gene. The two groups of selected cells were subsequently cultured for long periods up to 250 cell generations. Comparison of long-term GFP transgene integration and expression in these two groups of cells revealed that the K562 genome contains two types of transgene integration sites: i) abundant unstable sites that permit transcription but not long-term integration of the transgenes and thus eliminate the transgenes in 60-250 cell generations and ii) rare stable sites that permit both efficient transcription and long-term stable integration of the transgenes for at least 200 cell generations. Our results indicate that extinction of GFP expression with time is due at least in part to elimination of the gene from the host genome and not entirely to transcriptional silencing of the gene. However, long-term, stable expression of the transgene can be achieved in cells containing the transgene integrated into the rare, stable host sites. PMID- 11054550 TI - Phenol/cresol degradation by the thermophilic Bacillus thermoglucosidasius A7: cloning and sequence analysis of five genes involved in the pathway. AB - Bacillus thermoglucosidasius A7 degraded phenol at 65 degrees C via the meta cleavage pathway. Five enzymes used in the metabolism of phenol were cloned from B. thermoglucosidasius A7 into pUC18. Nine open reading frames were present on the 8.1kb insert, six of which could be assigned a function in phenol degradation using database homologies and enzyme activities. The phenol hydroxylase is a two component enzyme encoded by pheA1 and pheA2. The larger component (50kDa) has 49% amino acid identity with the 4-hydroxyphenylacetate hydroxylase of Escherichia coli, while the smaller component (19kDa) is most related (30% amino acid identity) to the styrene monoxygenase component B from Pseudomonas fluorescens. Both components were neccessary for activity. The catechol 2, 3-dioxygenase encoded by pheB has 45% amino acid identity with dmpB of Pseudomonas sp. CF600 and could be assigned to superfamily I, family 2 and a new subfamily of the Eltis and Bolin grouping. The 2-hydroxymuconic acid semialdehyde hydrolase (2HMSH), encoded by pheC, revealed the highest amino acid identity (36%) to the equivalent enzyme from Pseudomonas sp. strain CF600, encoded by dmpD. Based on sequence identity, pheD and pheE were deduced to encode the 2-hydroxypenta-2,4-dienoate hydratase (2HDH), demonstrating 45% amino acid identity to the gene product of cumE from Pseudomonas fluorescens and the acetaldehyde dehydrogenase (acylating) demonstrating 57% amino acid identity to the gene product of bphJ from Pseudomonas LB400. PMID- 11054551 TI - Expression of the bph genes involved in biphenyl/PCB degradation in Pseudomonas sp. KKS102 induced by the biphenyl degradation intermediate, 2-hydroxy-6-oxo-6 phenylhexa-2,4-dienoic acid. AB - The bph genes involved in PCB/biphenyl degradation in Pseudomonas sp. KKS102 are clustered as bphEGFA1A2A3BCDA4R. The bph genes are inducibly expressed in the presence of biphenyl. In order to understand the induction more fully, the inducer of bph gene expression was investigated. To identify the inducer molecule, we constructed four deletion mutants of the structural genes and analyzed the inducibility of the bphE gene in each mutant strain. In the wild type cell and the bphD deletion mutant, the levels of the bphE transcript were enhanced in the presence of biphenyl. On the other hand, in the bphA, bphB, and bphC deletion mutants, levels of the bphE transcript were not enhanced in the presence of biphenyl. These results demonstrated that the series of reactions catalyzed by biphenyl dioxygenase (BphA), dihydrodiol dehydrogenase (BphB), and 2, 3-dihydroxybiphenyl dioxygenase (BphC) are necessary to convert biphenyl to the inducer. It is known that these reactions convert biphenyl to 2-hydroxy-6-oxo 6-phenylhexa-2,4-dienoic acid (HOPDA), and it was found that the expression of the bph genes was induced by purified HOPDA. These results clearly indicate that HOPDA is the inducer of the bph genes in KKS102. PMID- 11054552 TI - Affinity selection of cDNA libraries by lambda phage surface display. AB - Bacteriophage lambda surface display was used to isolate cDNA clones encoding autoantigens recognized by sera from patients with Sjogren's syndrome (SS). We made cDNA libraries from human HeLa and HepG2 cells, using the expression vector lambdafoo. By repeating affinity selection of the libraries with the sera immobilized in microtiter wells, we isolated three clones that encode previously unknown antigens as well as four clones previously known as SS autoantigens. The newly identified autoantigens include TRK-fused gene product (TFG), survival motor neuron gene product (SMN) and pM5, which has a similarity to the metal binding domain of human fibroblast collagenase. Thus, the bacteriophage lambda surface display is powerful for isolating cDNA clones by affinity screening. PMID- 11054553 TI - Human protein tyrosine phosphatase-like gene: expression profile, genomic structure, and mutation analysis in families with ARVD. AB - The mouse protein tyrosine phosphatase-like gene (Ptpla) was recently cloned and data suggested that it plays a role in myogenesis and cardiogenesis. The human homologue (PTPLA) was mapped to chromosome 10p13-14, a region where we have mapped a locus responsible for arrhythmogenic right ventricular dysplasia (ARVD). As a positional candidate gene, we characterized PTPLA by determining its tissue expression, its genomic structure, and we also screened for mutations in the ARVD patients. Northern analysis demonstrated PTPLA is preferentially expressed in both adult and fetal heart. A much lower expression was detected in skeletal and smooth muscle tissues. Virtually no expression was observed in other tissues. The protein-encoding sequences of PTPLA consist of seven exons. A sequence variation (Lys64Gln) was found in all the affecteds in a large ARVD family. However, the same variant was also detected in normal control subjects (three alleles/100 chromosomes). Thus, the variant (Lys64Gln) is not responsible for ARVD in our family and is a benign polymorphism. Nevertheless, its tissue-specific expression in the developing and adult heart suggest PTPLA has a role in regulating cardiac development, differentiation, or other cellular events. The genomic structure and intragenic polymorphism of PTPLA should be useful for further clinical and genetic studies such as gene targeting of PTPLA. PMID- 11054554 TI - Redundant and non-functional guide RNA genes in Trypanosoma brucei are a consequence of multiple genes per minicircle. AB - The mitochondrial mRNA of the parasitic protozoa Trypanosoma brucei is extensively edited by the insertion, and occasional deletion, of uridine residues. The editing is mediated by over 200 guide RNAs (gRNAs) that are encoded in circular DNA molecules called minicircles. There are some 250 different types of minicircle, called classes, with each encoding several gRNAs. Sequencing of gRNAs and minicircles has revealed a surprising amount of both redundancy, where gRNAs from different minicircle classes edit exactly the same part of an mRNA, and non-functionality, where partial or no complementarity is found between gRNA and mRNA. How does this redundancy and non-functionality arise and persist? We propose the following. Minicircle classes that contain several functional gRNA genes can be lost from the population via drift and replaced by more minicircle classes that contain fewer functional gRNA genes, on the condition that the cells keep a full complement of functional gRNAs. We demonstrate this hypothesis in a computer simulation of a model of minicircle evolution. We show that this process leads to an increasing number of minicircle classes and inevitably to only one functional gRNA per minicircle. Moreover, we show that the genome contains more minicircle classes than is actually necessary for cell survival. We also analyse the available minicircle sequence data and conclude that T. brucei is at a transient stage in this process. In addition, ten new putative gRNAs have been discovered. PMID- 11054555 TI - Human allantoicase gene: cDNA cloning, genomic organization and chromosome localization. AB - Uric-acid-degrading enzymes (uricase, allantoinase, allantoicase, ureidoglycolate lyase and urease) were lost during vertebrate evolution and the causes for this loss are still unclear. We have recently cloned the first vertebrate allantoicase cDNA from the amphibian Xenopus laevis. Surprisingly, we have found some mammalian expressed sequence tags (ESTs) that show high similarity with Xenopus allantoicase cDNA. From a human fetal spleen cDNA library and adult kidney EST clone, we have obtained a 1790 nucleotide long cDNA. The 3' end of this sequence reveals a substantial high identity with the corresponding portion of Xenopus allantoicase cDNA. In contrast, at the 5' end the human sequence diverges from that of Xenopus; since no continuous open reading frame can be found in this region, the hypothetical human protein appears truncated at its N-terminus. We proposed that such a transcript could be due to an incorrect splicing mechanism that introduces an intron portion at the 5' end of human cDNA. Allantoicase cDNA is expressed in adult testis, prostate, kidney and fetal spleen. By comparison with available genomic sequences deposited in database, we have determined that the human allantoicase gene consists of five exons and spans 8kb. We have also mapped the gene in chromosome 2. PMID- 11054556 TI - Molecular characterization of human SUR2-containing K(ATP) channels. AB - The distribution of human sulfonylurea receptor-2 (SUR2)-containing K(ATP) channels was investigated using reverse transcriptase-polymerase chain reaction (RT-PCR). mRNA for SUR2B was detected in a variety of tissues including brain, skeletal, cardiac and smooth muscle, whereas SUR2A message was restricted to cardiac and skeletal muscle. An additional splice variant of SUR2 that lacked exon 17 was also identified by RT-PCR in tissues expressing both SUR2A and SUR2B or SUR2B alone. Quantification of RNA for SUR2 exon 17+ and SUR2 exon 17- splice variants using real-time Taqman PCR indicated differential levels of expression in brain, kidney, skeletal muscle, heart and small intestine. Interestingly, the SUR2 exon 17+ variant is the major species expressed in all tissues examined in this study. Each of the SUR2 splice variants transiently expressed with the inward rectifier Kir 6.2 formed functional K(ATP) channels in HEK 293 cells as assessed either by changes in DiBAC(4)(3) fluorescence responses or glyburide sensitive whole cell currents. Collectively, our findings demonstrate that various SUR2 splice variants have distinct expression patterns and can form functional K(ATP) channels. PMID- 11054557 TI - Genomic organization of mouse and human erythrocyte tropomodulin genes encoding the pointed end capping protein for the actin filaments. AB - Erythrocyte tropomodulin (E-Tmod), a globular protein of 359 residues, is highly expressed in the erythrocyte, heart and skeletal muscle. By binding to the N terminus of tropomyosin (TM) and actin, E-Tmod blocks the elongation and depolymerization of the actin filaments at the pointed end. In erythrocytes, the E-Tmod/TM complex contributes to the formation of the short actin protofilament, which in turn defines the geometry of the membrane skeleton. In juvenile mice, over-expression of E-Tmod is associated with dilated cardiomyopathy. We have previously cloned the human E-Tmod cDNA, identified its TM-binding region, and mapped its gene to chromosome 9q22. Through genomic library screening and PCR based genomic walking we have now cloned the mouse E-Tmod gene, whose coding region spans approximately 60kb containing nine exons and eight introns. The human E-Tmod gene obtained by PCR has an identical exon-intron organization. In sanpodo, a Tmod homologue in Drosophila, the exon boundaries are also conserved except that exons 2-5 and 6-7 are 'fused' and alternative splicing of two additional 5' exons and the 3' exons may give rise to several sanpodo isoforms. In a Tmod-like gene of C. elegans, exons 2-3 are 'fused', boundaries of exons 1, 7, 8, and 9 are conserved and exon/intron junctions of exons 4, 5 and 6 are shifted by a few residues. Analyses of 15 Tmod members from six species show no insertions or deletions of residues in the region of exons 6 and 7. A 5' rapid amplification of cDNA ends reveals that mouse E-Tmod transcripts obtained from embryonic stem cells, skeletal muscle and heart, but not smooth muscle, contain an additional 86bp untranslated cDNA sequence further upstream from exon 1. Thus, alternative promoters may provide a possible mechanism for tissue-specific expression and regulation of E-Tmod. This study is the first to report the exon organization of E-Tmod genes, which allows their regulation, manipulation, and disease relevance to be further investigated. PMID- 11054558 TI - Sp1 and Sp3 transactivate the RET proto-oncogene promoter. AB - The RET proto-oncogene plays an important role in the initiation and progression of tumors derived from the neural crest. The cis-regulatory elements responsible for RET basal promoter activity have not been identified. To characterize these elements, a RET promoter DNA fragment (-453 to +227bp) was fused to a luciferase reporter and introduced into TT, a neural crest-derived cell line. Sequential 5' deletions of the promoter revealed that optimal expression of the RET promoter in TT cells required only 70bp of sequence upstream of the transcription start site, and contains two Sp1 binding sites. DNase I footprinting, electrophoretic mobility shift analysis (EMSA), and supershift assays revealed that this region binds both Sp1 and its related protein, Sp3. Additionally, RET basal promoter activity was abrogated by removal of these Sp1/Sp3 binding sites. The proximal two GC boxes were sufficient to allow transactivation of the RET promoter in Drosophila SL2 cells. Sp3 expression in these cells caused an additional activation of the promoter. These results demonstrate that the transactivation of the RET promoter within a neural crest-derived cell line is dependent on Sp1 and Sp3. PMID- 11054559 TI - 4SR, a novel zinc-finger protein with SR-repeats, is expressed during early development of Xenopus. AB - The protein C4SR contains two cysteine(4) (C(4)) zinc-finger motifs at its amino terminus, a stretch of acidic residues in the middle and a series of serine arginine (SR) repeats at its carboxyl terminus. A cDNA clone encoding the zinc finger domain was first selected from a Xenopus laevis oocyte expression library on the basis of the ability of the fusion protein to stably bind an RNA probe. The mRNA encoding C4SR is expressed during oogenesis, and the protein is present at a constant level in oocytes and early embryos. The C4SR protein is expressed in transcriptionally active erythroblasts but not in transcriptionally inert mature erythrocytes. An epitope-tagged C4SR protein, expressed in oocytes, associates with nascent transcripts at many loci in lampbrush chromosomes and is absent from storage particles (snurposomes) containing the normally recognized complement of RNA splicing components. It is likely that C4SR is involved in pre mRNA transcription/packaging rather than in exon splicing. The zinc-finger motif, present as two copies in C4SR, is also present in a range of transcription associated proteins. We suggest the descriptor (DW)C(4), in which DW refers to the invariant aspartic acid (D)/tryptophan (W) dipeptide that precedes the first cysteine residue, for this distinctive zinc-finger structure. PMID- 11054560 TI - A zebrafish vitellogenin gene (vg3) encodes a novel vitellogenin without a phosvitin domain and may represent a primitive vertebrate vitellogenin gene. AB - By analysis of zebrafish EST (expressed sequence tag) clones from an adult cDNA library, we have identified 44 clones, about 11% of the adult EST clones, encoding vitellogenins. These vitellogenin EST clones have been derived from at least seven distinct vitellogenin genes. One of the largest vitellogenin cDNA clones, vg3, and its 5' extended clone isolated by 5' RACE (rapid amplification of cDNA ends)-PCR, have been sequenced completely. The deduced complete sequence includes a predicted mature vitellogenin of 1233 amino acids and a truncated signal peptide of 18 amino acids. Interestingly, the predicted vitellogenin has no polyserine phosvitin domain. The lack of the phosvitin domain was confirmed by isolation and sequencing of the vg3 genomic region. Phylogenetic analysis indicates that the phosvitinless vitellogenin is an intermediate between invertebrate vitellogenins and all known vertebrate vitellogenins, and thus may represent a primitive vertebrate vitellogenin. Like other vitellogenins in vertebrates, the phosvitinless vitellogenin is also synthesized mainly in the liver and weakly in the intestine. PMID- 11054561 TI - C11orf21, a novel gene within the Beckwith-Wiedemann syndrome region in human chromosome 11p15.5. AB - A novel gene, C11orf2, was identified by BLAST search in the human chromosome 11p15.5 region potentially responsible for Beckwith-Wiedemann Syndrome (BWS) and some cancers. Two cDNA clones with different sizes were obtained, which share a potential ORF of 399bp and are different in their 3' untranslated regions. This gene was revealed to be expressed exclusively in human heart and in almost no other tissues examined by northern blotting. Two transcripts of different sizes, 0.9 and 3.1kb, were identified in heart, consistent with the length of the two cDNA clones. The gene shows biallelic expression (non-imprinted) in fetal liver, although it is located in the imprinted domain of 11p15.5. C11orf21 codes a protein of 132 amino acids as proved by the expression of C11orf21-EGFP fusion protein in cultured cells. The EGFP-fusion protein expressed in cultured cells localized mainly in the cytoplasm. PMID- 11054562 TI - FK506-binding protein-type peptidyl-prolyl cis-trans isomerase from a halophilic archaeum, Halobacterium cutirubrum. AB - The halophilic archaeum, Halobacterium cutirubrum, has been shown to have a cyclophilin-type peptidyl-prolyl cis-trans isomerase (PPIase). Because most archaeal genomes studied only have genes for FK506-binding proteins (FKBPs) as a PPIase, it has been unclear whether H. cutirubrum has an FKBP-type PPIase or not. In the present study, a gene encoding an FKBP-type PPIase was cloned from genomic DNA of H. cutirubrum and then sequenced. This FKBP was deduced to be composed of 303 amino acid residues with a molecular mass of 33.3kDa. Alignment of its amino acid sequence with those of other reported FKBPs showed that it contained two insertion sequences in the regions corresponding to the bulge and flap of human FKBP12, which are common to archaeal FKBPs. Its C-terminal amino acid sequence was approximately 130 amino acids longer than the FKBPs of Methanococcus thermolithotrophicus and Thermococcus sp. KS-1. Among the 14 conserved amino acid residues that form the FK506 binding pocket, only three were found in this FKBP. This gene was expressed as a fusion protein with glutathione S-transferase (GST) in Escherichia coli, and the N-terminal GST portion was removed by protease digestion. The purified recombinant FKBP showed a weak PPIase activity with a low sensitivity to FK506. This FKBP suppressed aggregation of the unfolded protein. PMID- 11054563 TI - Snail/slug family of repressors: slowly going into the fast lane of development and cancer. AB - The existence of homologous genes in diverse species is intriguing. A detailed comparison of the structure and function of gene families may provide important insights into gene regulation and evolution. An unproven assumption is that homologous genes have a common ancestor. During evolution, the original function of the ancestral gene might be retained in the different species which evolved along separate courses. In addition, new functions could have developed as the sequence began to diverge. This may also explain partly the presence of multipurpose genes, which have multiple functions at different stages of development and in different tissues. The Drosophila gene snail is a multipurpose gene; it has been demonstrated that snail is critical for mesoderm formation, for CNS development, and for wing cell fate determination. The related vertebrate Snail and Slug genes have also been proposed to participate in mesoderm formation, neural crest cell migration, carcinogenesis, and apoptosis. In this review, we will discuss the Snail/Slug family of regulators in species ranging from insect to human. We will present the protein structures, expression patterns, and functions based on molecular genetic analyses. We will also include the studies that helped to elucidate the molecular mechanisms of repression and the relationship between the conserved and divergent functions of these genes. Moreover, the studies may enable us to trace the evolution of this gene family. PMID- 11054564 TI - Characterization of the hCTR1 gene: genomic organization, functional expression, and identification of a highly homologous processed gene. AB - The human hCTR1 gene was originally identified by its ability to complement a yeast mutant deficient in high-affinity copper uptake (Zhou, B., Gitschier, J., 1997. A human gene for copper uptake identified by complementation in yeast. Proc. Natl. Acad. Sci. USA 94, 7481-7486). Here, we have determined the DNA sequence of the exon-intron borders of the hCTR1 structural gene and report that the coding sequence is disrupted by three introns, all of which comply with the GT/AG rule. Furthermore, human fibroblasts, transfected with hCTR1 cDNA, were shown to have a dramatically increased capacity for (64)Cu uptake, indicating that the hCtr1 protein is functional in copper uptake in human cells. In contrast, no evidence was found for involvement of the hCTR2 gene product in copper uptake. Finally, we have identified a highly homologous processed pseudogene, hCTR1psi, which was localized to chromosome 3q25/26. The processed gene was found to be transcribed, but due to a frame shift mutation, it only had the potential to encode a truncated protein of 95 amino acid residues, and cells transfected with hCTR1psi DNA showed no increase of (64)Cu uptake. PMID- 11054565 TI - p190-A, a human tumor suppressor gene, maps to the chromosomal region 19q13.3 that is reportedly deleted in some gliomas. AB - To date, two distinct genes coding for Ras GAP-binding phosphoproteins of 190kDa, p190-A and p190-B, have been cloned from mammalian cells. Rat p190-A of 1513 amino acids shares 50% sequence identity with human p190-B of 1499 amino acids. We have previously demonstrated, using rat p190-A cDNA, that full-length p190-A is a tumor suppressor, reversing v-Ha-Ras-induced malignancy of NIH 3T3 cells through both the N-terminal GTPase (residues 1-251) and the C-terminal Rho GAP (residues 1168-1441) domains. Here we report the cloning of the full-length human p190-A cDNA and its first exon covering more than 80% of this protein, as well as its chromosomal mapping. Human p190-A encodes a protein of 1514 amino acids, and shares overall 97% sequence identity with rat p190-A. Like the p190-B exon, the first exon of p190-A is extremely large (3.7 kb in length), encoding both the GTPase and middle domains (residues 1-1228), but not the remaining GAP domain, suggesting a high conservation of genomic structure between two p190 genes. Using a well characterized monochromosome somatic cell hybrid panel, fluorescent in situ hybridization (FISH) and other complementary approaches, we have mapped the p190-A gene between the markers D19S241E and STD (500 kb region) of human chromosome 19q13.3. Interestingly, this chromosomal region is known to be rearranged in a variety of human solid tumors including pancreatic carcinomas and gliomas. Moreover, at least 40% glioblastoma/astrocytoma cases with breakpoints in this region were previously reported to show loss of the chromosomal region encompassing p190-A, suggesting the possibility that loss or mutations of this gene might be in part responsible for the development of these tumors. PMID- 11054566 TI - Evidence that dim1 associates with proteins involved in pre-mRNA splicing, and delineation of residues essential for dim1 interactions with hnRNP F and Npw38/PQBP-1. AB - The small evolutionarily conserved protein Dim1p/hDim1/Dib1p/DML-1 was initially defined as a factor essential for progression through the G2/M transition, and shown to be required to maintain the steady state level of a component of the fission yeast anaphase promoting complex/cyclosome. More recently, Dib1p has been defined as a component of the U4/U6.U5 tri-snRNP, required for pre-mRNA splicing. To investigate the mechanism(s) of Dim1 function, reiterative two-hybrid screening was performed to identify interacting proteins. Proteins thus identified were solely those involved in pre-mRNA splicing or related functions, and one partner induced a striking synthetic phenotype when co-expressed with hDim1 in mammalian cells. Saturating alanine scanning mutagenesis of Dim1 allowed delineation of amino acids essential for its ability to interact with its defined partners: mapping these residues on the structural coordinates of hDim1 defined an interactive sector of the protein. Finally, depletion studies have recently shown that Dim1 function is essential for pre-mRNA splicing in yeast. We find that elimination of DML-1 expression in C. elegans by RNA interference leads to embryonal lethality during gastrulation, marked by a failure to correctly express early zygotic transcripts. These results parallel the arrest phenotypes associated with global disruption of zygotic gene expression, suggesting that Dim1 proteins maintain an essential function in gene expression in higher eukaryotes. PMID- 11054567 TI - Molecular cloning and characterization of a protein tyrosine phosphatase enriched in testis, a putative murine homologue of human PTPMEG. AB - Protein tyrosine phosphorylation is regulated by protein tyrosine kinase and protein tyrosine phosphatase activities. These two counteracting proteins are implicated in cell growth and transformation. Using polymerase chain reaction with degenerate primers, we have identified a novel mouse protein tyrosine phosphatase (PTP). This cDNA contains a single open reading frame of the predicted 926 amino acids. Those predicted amino acids showed significant identity with human megakaryocyte protein-tyrosine phosphatase by 91% in nucleotide sequences and 94% in amino acid sequences. We have identified that expression of this PTP is highly enriched in the testis in mouse and human and has been termed here as a 'testis-enriched phosphatase' (TEP). Northern analysis detected two mRNA species of 3.7 and 3.2kb for this PTP in mouse testis and the expression of TEP is regulated during development. The recombinant phosphatase domain possesses protein tyrosine phosphatase activity when expressed in Escherichia coli. Immunohistochemical analysis of the cellular localization of TEP on mouse testis sections showed that this PTP is specifically expressed in spermatocytes and spermatids within seminiferous tubules, suggesting an important role in spermatogenesis. PMID- 11054568 TI - Puromycin resistance (pac) gene as a selectable marker in vaccinia virus. AB - The antibiotic puromycin, an inhibitor of protein synthesis, was shown to inhibit vaccinia virus (VV) replication. We evaluated the use of puromycin-resistance (pac) gene as a selectable marker in VV. A recombinant vaccinia virus expressing pac (VV-pac) under the control of a viral early/late promoter was constructed and characterized. VV-pac grew in the presence of puromycin at concentrations that were inhibitory for the parental VV and toxic for the cells. Isolation of recombinant VV usually relies on plaque purification under selective conditions. Because virus plaquing was not feasible under inhibitory puromycin concentration, a protocol based on serial passage of virus was devised. The usefulness of this procedure in selecting pac expressing viruses was tested by isolating a recombinant VV. PMID- 11054569 TI - Rab6c, a new member of the rab gene family, is involved in drug resistance in MCF7/AdrR cells. AB - A new Rab6 homolog cDNA, Rab6c, was discovered by a hypermethylated DNA fragment probe that was isolated from a human multidrug resistant (MDR) breast cancer cell line, MCF7/AdrR, by the methylation sensitive-representational difference analysis (MS-RDA) technique. Rab6c was found to be under-expressed in MCF7/AdrR and MES-SA/Dx5 (a human MDR uterine sarcoma cell line) compared with their non MDR parental cell lines. MCF7/AdrR cells expressing the exogenous Rab6c exhibited less resistance to several anti-cancer drugs, such as doxorubicin (DOX), taxol, vinblastine, and vincristine, than the control cells containing the empty vector. Flow cytometry experiments confirmed that the transfectants' diminished resistance to DOX was caused by increased drug accumulation induced by the exogenous Rab6c. These results indicate that Rab6c is involved in drug resistance in MCF7/AdrR cells. PMID- 11054570 TI - Expression of cyclin E in endomitotic silk-gland cells from mulberry silkworm. AB - The silk glands of mulberry silkworm Bombyx mori are endoreplicating tissues in which the genomic DNA undergoes multiple rounds of replication without mitosis and nuclear division. In the absence of normal mitotic division, the cell cycle essentially alternates between the G1 and S phases. Cyclin E is crucial for the G1/S transition in both mitotic and endoreplicating cycles. We have cloned and characterized cyclin E (cyclin box) from B. mori, which is nearly identical to the Drosophila cyclin E box except for an insertion of 21 amino acids. Two distinct cyclin E transcripts (1.7 and 2.1 kb) were detected in the silk-gland cells of B. mori and in the B. mori-derived embryonic cell line, BmN. Using anti Cyclin E antibodies two protein bands of 52 and 44kDa were detected in silk glands and BmN cells at comparable levels. Both BmN- and the silk-gland cells showed the presence of the interacting kinase Cdk2. Transcripts of the mitotic cyclin, cyclin B, were barely detectable in the endoreplicating silk-gland cells and amounted to only 4-7% of that seen in the mitotically dividing BmN cells. The near absence of cyclin B transcripts and the abundant expression of cyclin E in the silk glands correlate well with the alternation of only G1 and S phases without the intervening mitosis in these cells. PMID- 11054571 TI - Phylogenetic analysis of teneurin genes and comparison to the rearrangement hot spot elements of E. coli. AB - Teneurins are a novel family of transmembrane proteins conserved between invertebrates and vertebrates. There are two members in Drosophila, one in C. elegans and four members in mouse. Here, we describe the analysis of the genomic structure of the human teneurin-1 gene. The entire human teneurin-1 (TEN1) gene is contained in eight PAC clones representing part of the chromosomal locus Xq25. Interestingly, many X-linked mental retardation syndromes (XLMR) and non-specific mental retardation (MRX) are mapped to this region. The location of the human TEN1 together with the neuronal expression makes TEN1 a candidate gene for XLMR and MRX. We also identified large parts of the human teneurin-2 sequence on chromosome 5 and sections of human teneurin-4 at chromosomal position 11q14. Database searches resulted in the identification of ESTs encoding parts of all four human members of the teneurin family. Analysis of the genomic organization of the Drosophila ten-a gene revealed the presence of exons encoding a long form of ten-a, which can be aligned with all other teneurins known. Sequence comparison and phylogenetic trees of teneurins show that insects and vertebrates diverged before the teneurin ancestor was duplicated independently in the two phyla. This is supported by the presence of conserved intron positions between teneurin genes of man, Drosophila and C. elegans. It is therefore not possible to class any of the vertebrate teneurins with either Drosophila Ten-a or Ten-m. The C-terminal part of all teneurins harbours 26 repetitive sequence motifs termed YD repeats. YD-repeats are most similar to the repeats encoded by the core of the rearrangement hot spot (rhs) elements of Escherichia coli. This makes the teneurin ancestor a candidate gene for the source of the rhs core acquired by horizontal gene transfer. PMID- 11054572 TI - Genomic analysis and functional expression of canine dopamine D2 receptor. AB - Dopamine D2 receptor (DRD2) is one of the five dopamine receptors with seven transmembrane domains that are coupled to the G protein. We have cloned and characterized the genomic and cDNA sequences of the canine DRD2 gene, which are 12.7 and 2.7 kb in size, respectively. The genomic DNA is composed of seven exons and six introns, encoding a 443 amino acid protein with 95% amino acid identity to other mammalian D2 receptors. A length polymorphism was detected in intron 3 of the receptor gene. We also characterized alternatively spliced forms of DRD2 cDNAs, DRD2L and DRD2S. They showed a higher level of expression in midbrain and thalamus. The ratio between the long and short form is similar in RT-PCR reaction. In human and rodent, the same two spliced forms are known to be coupled to G(i)-type heterotrimeric GTP binding protein, thereby opening an inwardly rectifying potassium channel, GIRK1. When the canine DRD2L and DRD2S were heterologously expressed in Xenopus oocytes, both forms activated GIRK1 potassium channels through coupling with G(i) protein. This activation was dose-dependent, demonstrating its ligand specificity. PMID- 11054573 TI - Characterization of the human tubulin tyrosine ligase-like 1 gene (TTLL1) mapping to 22q13.1. AB - This paper reports the characterization of the human tubulin tyrosine ligase-like 1 gene (TTLL1), which maps to the chromosome region 22q13.1 and has been partially duplicated on three other acrocentric chromosomes: 13, 15 and 21. We describe the complete cDNA, TTLL1a, coding for the putative 423 amino acid long TTLL1 and alternative transcripts coding for truncated TTLL1. Likely TTLL1a corresponds to the 1.8 kb transcript that was detected in a wide range of tissues and has a stronger expression in heart, brain and testis. A 4.8 kb transcript was found only in brain tissues. We present an interspecies sequence comparison, revealing three conserved domains, named TTLD1, TTLD2 and TTLD3, that are specific to the TTLs and TTL-like proteins. PMID- 11054574 TI - Sequencing and expression analysis of the serine protease gene cluster located in chromosome 19q13 region. AB - The human kallikrein gene cluster, located in the chromosome band 19q13, contains several tissue-specific serine protease genes including the prostate-specific KLK2, KLK3 and prostase genes. To further characterize the gene cluster, we have mapped, sequenced, and analyzed the genomic sequence from the region. The results of EST database searches and GENSCAN gene prediction analysis reveal 13 serine protease genes and several pseudogenes in the region. Expression analysis by RT PCR indicates that most of these protease genes are expressed only in a subset of the 35 different normal tissues that have been examined. Several protease genes expressed in skin show higher expression levels in psoriatic lesion samples than in non-lesional skin samples from the same patient. This suggests that the imbalance of a complex protease cascade in skin may contribute to the pathology of disease. The proteases, excluding the kallikrein genes, share approximately 40% of their sequences suggesting that the serine protease gene cluster on chromosome 19q13 arose from ancient gene duplications. PMID- 11054575 TI - Yeast Pdr13p and Zuo1p molecular chaperones are new functional Hsp70 and Hsp40 partners. AB - The deletion of the TOM1 gene encoding a putative ubiquitin ligase causes a temperature sensitive cellular growth in Saccharomyces cerevisiae. The arrested cells exhibit pleiotropic defects in nuclear division, maintenance of nuclear structure and heat stress responses. We previously identified a zuo1 mutation as an extragenic suppressor of the tom1 mutant. ZUO1 encodes a DnaJ-related Hsp40. Here we show that a recessive cold sensitive mutation in PDR13 coding for an Hsp70 suppressed the tom1 mutation. The pdr13 deletion mutant was sensitive to high osmolarity, just like the zuo1 deletion mutant. A zuo1 pdr13 double deletion mutant did not show additive phenotypes. Furthermore, a tagged-Zuo1p was co immunoprecipitated with a tagged-Pdr13p. Taken together, we propose that Pdr13p and Zuo1p are a new pair of Hsp70:Hsp40 molecular chaperones. In addition, Pdr13p co-sedimented with translating ribosomes and this association was independent of the presence of Zuo1p. PMID- 11054576 TI - Molecular cloning of an Atlantic salmon nucleoside diphosphate kinase cDNA and its pattern of expression during embryogenesis. AB - To gain insight into the process of development in Atlantic salmon (Salmo salar), we sought to identify genes that were differentially expressed at gastrulation. A polymerase chain reaction-based differential screening strategy allowed for the isolation of an Atlantic salmon nucleoside diphosphate kinase cDNA (nm23). Structural characterisation showed a high degree of homology with a large number of previously isolated nucleoside diphosphate kinases (NM23s), both prokaryote and eukaryote, though it represents the first teleost nucleoside diphosphate kinase identified. Highest similarities were found with the type 1 and type 2 NM23 isoforms of mammals. Phylogenetic analysis indicates that the duplication event that gave rise to these isoforms occurred after the splitting of tetrapods and fish, suggesting that the salmon NM23 represents a more ancestral isoform. The position of the salmon sequence on the phylogenetic tree indicates that the salmon genome is expected to have at least three copies of genes from the nm23 gene family. Northern blot analysis showed a single transcript of approximately 0.7 kb in both embryonic and adult tissues. Examination of the temporal pattern of expression of salmon nucleoside diphosphate kinase during embryonic development revealed that this gene is first expressed at the time of gastrulation. Nucleoside diphosphate kinases are thought to have a vital role in regulatory processes such as signal transduction, proliferation and differentiation. Taken together, these results suggest that nucleoside diphosphate kinases have an important role to play in early embryogenic development in vertebrates. PMID- 11054577 TI - Cloning, expression and purification of recombinant cotton rat interleukin-5. AB - The coding sequence of the hispid cotton rat (Sigmodon hispidus) interleukin-5 (IL-5) was isolated by a combination of reverse transcription (RT)-PCR and RACE protocols from concanavalin A stimulated spleen cells. The open reading frame of 399 bp encodes a polypeptide of 132 amino acids. Comparison with the rat, mouse, gerbil and human counterparts revealed 88, 88, 87 and 75% identity at the nucleotide level and 88, 90, 89 and 70% at the amino acid level, respectively. The entire coding sequence, minus the putative signal peptide sequence, was inserted into an inducible Escherichia coli expression vector. The recombinant protein possessed an expected molecular mass of 14kDa and was located in bacterial inclusion bodies. A purification scheme under reducing and denaturing conditions followed by subsequent successive dialysis steps led to the recovery of a recombinant dimeric cotton rat IL-5. The biological activity of the recombinant protein was demonstrated in a murine cell line proliferation assay. This activity was specifically inhibited by rat monoclonal antibodies directed against mouse IL-5. Together with specific antibodies that can be generated easily, cotton rat IL-5 constitutes a useful tool for extending the use of the cotton rat animal model in the study of various human pathogens. PMID- 11054578 TI - Ty1-copia retrotransposon-like elements in chickpea genome: their identification, distribution and use for diversity analysis. AB - Ty1-copia retrotransposon-like elements were amplified from Cicer species using primers derived from the conserved region of the reverse transcriptase gene. Two fragments, of size approximately 280bp and approximately 650 bp, were obtained, which on sequencing showed homology for the Ty1-copia reverse transcriptase region. Interestingly, the approximately 650 bp fragment showed two reverse transcriptase regions, one from Ty1-copia and the other from Tto1 element fused together. The copy number was high in the cultivated Cicer arietinum genome compared with the wild Cicer reticulatum. Genetic diversity among the Cicer species was investigated using the conserved primers which grouped the wild species and the cultivated C. arietinum separately. PMID- 11054579 TI - Diploid/triploid mosaic placenta with fetus. Towards A better understanding of 'partial moles'. AB - We describe a case of partial molar change in a placenta that was associated with a normal female fetus who died in utero. The analysis of molar and normal placental tissue, as well as the karyotypic study of amnionic fluid indicate a complex origin of this conceptus. We review the possible mechanisms leading to this pregnancy and the general topic of partial hydatidiform mole. The formation of moles is complex and it is not easily divisible into so-called partial and complete hydatidiform moles. Rather, individual genetic study is needed to make an accurate diagnosis because macroscopic or microscopic examination alone fails to assess the complexity of these entities. PMID- 11054580 TI - Non-nutritive sucking in the healthy pre-term infant. AB - In the infant, sucking behaviour is one of the first coordinated muscular activities. It is under the control of the brainstem. In utero sucking is observed from 13 weeks' gestation. The healthy full-term newborn infant has a very stable rhythm of alternating bursts of sucking with pauses in between. The non-nutritive sucking (NNS) patterns of 58 low-risk prematurely born infants (gestational ages between 26 and 35 weeks) were studied using a specially designed computer-based method that analyses and quantifies NNS. A total of 183 observations were analysed. The typical NNS pattern with bursts of sucking activity separated by quiescent periods was recorded in all infants studied and was already present before 30 weeks of gestation. A gradual change over time of their NNS was seen. With increased maturation the sucking activity, sucking frequency, amplitude and burst duration all increased, while the variability of the sucking frequency and the duration of the intervals between bursts declined. Post-menstrual age (PMA) was the dominant predictor of this result but gender, state of activity and weight also influenced it to some extent. Girls had more sucking activity and a higher sucking frequency than boys. State of activity affected the stability of the rhythm. The weight of the infant influenced both sucking activity and duration of separate bursts. PMID- 11054581 TI - Serial measurements of serum human placental lactogen (hPL) and serial ultrasound examinations in the evaluation of fetal growth. AB - Serial serum hPL measurements and serial ultrasound fetometry were compared in the evaluation of fetal growth by relating these two parameters to size at birth and to clinical factors known to influence size at birth. The data were from a prospective study of 1000 consecutive pregnant women considered to be at risk for fetal growth retardation with retrospective analysis. Serum hPL was measured by radioimmunoassay and fetal weight estimated by ultrasound every 3 weeks during the last trimester. hPL values were expressed as multiples of the median (MoM) and linear regression analysis of the hPL MoM values was carried out for each pregnancy to find the slope of the line (hPL-slope); at least 3 serum hPL values were required. The estimated fetal weight and weight-for-age at birth was expressed in Z-scores. The individual intrauterine growth velocity was calculated by regression analysis and expressed as change in Z-score for 12 weeks. At least two ultrasound measurements over an interval of at least 42 days were used to estimate the fetal growth velocity. In 588 women the file was complete. The main outcome measures were the individual mean hPL, hPL-slope, fetal growth velocity, birth weight deviation, smoking in pregnancy and diagnosis of preeclampsia. A significant correlation was found between the hPL-slope and the intrauterine fetal growth velocity (r=0.34), and between hPL-slope and birth weight deviation (r=0.32). Mean hPL was correlated to birth weight deviation (r=0.27), but only very weakly to intrauterine growth velocity (r=0.08). hPL-slope and intrauterine growth velocity independently predicted birth weight deviation. Heavy smoking which was stopped before the third trimester was not associated with low intrauterine growth velocity, but with a low hPL-slope. Preeclampsia was associated with a trend towards low and decreasing hPL and with an increasing intrauterine growth velocity and birth weight deviation. In conclusion the rate of change of serial hPL measurements correlated well to intrauterine fetal growth velocity in the third trimester as estimated by ultrasound and to the deviation in birth weight, but hPL seems to have a separate physiological significance, since it did not pick up when smoking was stopped and growth velocity was normalised and it did not at all detect the increased growth associated with preeclampsia. PMID- 11054582 TI - The effects of pre- and post-natal sunlight exposure on human growth: evidence from the Southern Hemisphere. AB - Several recent studies have reported a causal association between stature and month of birth. Perinatal exposure to sunlight has been suggested as the principal factor underlying this relationship, although the mechanisms involved remain a matter of debate. The longitudinal design of the present study allowed us to directly test the influence of perinatal sunlight exposure (and other meteorological and behavioural factors) on body size at birth and at regular intervals up to age 26. The findings confirmed that pre-natal sunlight is one of the most significant determinants of height. However, the trimester of greatest influence differs depending on the age at which study members were measured. PMID- 11054583 TI - Ingestive behavior and obesity. PMID- 11054584 TI - Eating behavior: lessons from the real world of humans. AB - Food intake by normal humans has been investigated both in the laboratory and under free-living conditions in the natural environment. For measurement of real world intake, the diet-diary technique is imperfect and tends to underestimate actual intakes but it appears to be sensitive, can detect subtle influences on eating behavior, and produces reliable and valid measures. Research studies in the real world show the multivariate richness of the natural environment, which allows investigation of the complexities of intake regulation, and even causation can be investigated. Real-world research can overcome some of the weaknesses of laboratory studies, where constraints on eating are often removed or missing, facilitatory influences on eating are often controlled or eliminated, the importance of variables can be overestimated, and important influences can be missed because of the short durations of the studies. Real-world studies have shown a wide array of physiologic, psychological, and social variables that can have potent and immediate effects on intake. Compensatory mechanisms, including some that operate with a 2- to 3-d delay, adjust for prior excesses. Heredity affects all aspect of food-intake regulation, from the determination of body size to the subtleties of the individual preferences and social proclivities and the extent to which environmental factors affect the individual. Hence, real-world research teaches valuable lessons, and much more is needed to complement laboratory studies. PMID- 11054585 TI - The controls of eating: a shift from nutritional homeostasis to behavioral neuroscience. AB - The experimental analysis of the controls of eating has undergone a paradigmatic shift in the past decade. Instead of seeing meals as a problem of how intake serves metabolism and nutritional homeostasis, meals are now seen as a problem in behavioral neuroscience. The major developments underlying this significant change are behavioral, neurological, and theoretical. Behavioral analysis has shown that a central pattern generator in the caudal brainstem organizes eating movements and that the size of a liquid meal is determined by the number and size of clusters of licking. Neurologic analysis has shown that eating is under orosensory positive-feedback control and postingestive, preabsorptive, negative feedback control. These feedback controls are activated by food ingested during a meal. The sensory information of the feedbacks is carried by afferent fibers that project to the caudal brainstem. The new theory is based on the fact that the feedback controls are stimulated by food acting directly on mucosal receptors along the gastrointestinal tract, from the mouth to the end of the small intestine. Thus, they are referred to as direct controls, and the caudal brainstem is sufficient for organizing their action. All other controls, such as metabolic, rhythmic, and ecologic, that do not contact the mucosal receptors are indirect controls. Indirect controls act by modulating the potency of the central effects of the direct controls, and they require the forebrain and its reciprocal connections with the caudal brainstem for their control of eating and meal size. PMID- 11054586 TI - Thirst. AB - The homeostasis of body fluid traditionally is viewed as involving the regulation of its osmolality and of blood volume. However, the control of thirst is more complex than can be described in a two-factor model, and consideration of plasma sodium concentration and of arterial blood pressure also must be included in the discussion. This review is organized around those four variables and focuses on the seven distinct signals that appear to influence water intake in rats. These signals include four that are excitatory for thirst: increased plasma osmolality detected by cerebral osmoreceptors, decreased blood volume presumably detected by cardiac stretch receptors, increased circulating levels of angiotensin II detected by angiotensin II receptors in the subfornical organ, and increased gastric sodium load apparently detected by putative sodium receptors in the abdominal viscera. There also appear to be three signals that inhibit thirst: decreased plasma osmolality detected by cerebral osmoreceptors, increased arterial blood pressure detected by arterial baroreceptors, and increased gastric water load apparently detected by putative sodium receptors in the abdominal viscera. PMID- 11054587 TI - Molecular neurobiology of ingestive behavior. AB - The concepts and tools of molecular biology may be applied to almost any component of the animal involved in ingestion, but two categories of model system are particularly relevant for molecular analysis: homeostatic regulation of neuropeptide expression in the hypothalamus and neuronal plasticity underlying persistent changes in ingestive behavior. Molecular approaches to these models are reviewed, focusing on our strategy for analyzing conditioned taste aversion learning. Three questions must be answered: Where do the long-term changes occur within the distributed neural network that mediates feeding? This answer reveals the site of neuronal restructuring mediated by gene expression. When does the transition occur from short-term expression to long-term persistence of the change in behavior? This transition reveals the critical time of gene expression. What genes are expressed during the change in behavior? The expression of thousands of genes in discrete subpopulations of cells is likely to be required during critical periods of neuronal restructuring. The identification of these genes is a general challenge for molecular neurobiology. The analysis of ingestive behavior can profit from molecular tools, but ingestion also provides informative models that elucidate the principles of time- and neuron-specific gene expression mediating complex behaviors. PMID- 11054588 TI - Norepinephrine and the control of food intake. AB - The focus of the present review is to reconsider the role of endogenous norepinephrine (NE) in brain, specifically within the hypothalamic paraventricular nucleus (PVN), with regard to its potential role in eliciting eating or satiety. The PVN is innervated by NE fibers and is a site at which infusion of exogenous NE elicits eating at low doses. Two subtypes of alpha adrenergic receptors within the PVN exert antagonistic actions on eating in the rat: activation of PVN alpha(2)-adrenoceptors increases eating, whereas activation of PVN alpha(1)-adrenoceptors suppresses eating. Pharmacologic manipulations that elevate NE can increase or decrease food intake, depending on the site and type of NE manipulation. Certain antiobesity drugs may act to reduce eating via release of NE and subsequent activation of alpha(1)-adrenoceptors. The PVN exhibits a reliable rhythm in the secretion of endogenous NE over the dark and-light cycle, and this rhythm may interact with changes in numbers of PVN alpha(1)- and alpha(2)-adrenoceptors to modulate eating during the dark-and-light cycle. Push-and-pull and microdialysis studies indicate that NE secretion is strongly associated with eating, particularly at the start of the dark phase. The present review considers potential interactions of NE with substances such as leptin and neuropeptide Y that alter eating. PMID- 11054589 TI - Hypothalamic dopamine and serotonin in the regulation of food intake. AB - Because daily food intake is the product of the size of a meal and the frequency of meals ingested, the characteristic of meal size to meal number during a 24-h light-dark cycle constitutes an identifiable pattern specific to normal states and obesity and that occurs during early cancer anorexia. An understanding of simultaneous changes in meal size and meal number (constituting a change in feeding patterns) as opposed to an understanding of only food intake provides a more insightful dynamic picture reflecting integrated behavior. We have correlated this to simultaneous changes in dopamine and serotonin concentrations and to their postsynaptic receptors, focusing simultaneously on two discrete hypothalamic food-intake-related nuclei, in response to the ingestion of food. The relation between concentrations of dopamine and serotonin limited to the lateral hypothalamic area (LHA) and the ventromedial nucleus (VMN) as they relate to the influence of meal size and meal number during the hyperphagia of obesity and anorexia of cancer as measured in our experiments are discussed. Based on these data, conceptual models are proposed concerning: 1) an "afferent-efferent neurotransmitter unit," with facilitatory or inhibitory neuropeptide properties to generate an appropriate neuroendocrine and neuronal response that ultimately modifies food intake; 2) initiation and termination of a meal, thereby determining the number and size of a meal under normal conditions; and 3) a schema integrating the onset mechanism of cancer anorexia. Nicotine is used as a tool to further explore the relation of meal size to meal number, with a focus on simultaneous changes in dopamine and serotonin concentrations in the LHA and VMN with the onset of acute anorexia of nicotine infusion and acute hyperphagia of nicotine cessation. Data concerning the role of sex-related hormones on dopamine and serotonin with regard to the LHA and VMN in relation to the modulation of food intake are also presented. PMID- 11054590 TI - Cholecystokinin and satiety: current perspectives. AB - In the almost 30 years since the ability of peripheral administration of the brain/gut peptide cholecystokinin (CCK) to inhibit food intake was first demonstrated, significant progress in our overall understanding of the role of CCK in ingestive behavior has been made. A physiologic role for endogenous CCK in the control of meal size has been demonstrated and sites and mechanisms of action for CCK in food intake have been investigated. Recent work has uncovered roles for the CCK satiety pathway in the mediation of the feeding modulatory actions of estradiol, insulin, and leptin. The availability of the Otsuka Long Evans Tokushima Fatty (OLETF) rat, a strain lacking CCK(A) receptors, provides a unique model for the study of how deficits in a within-meals satiety signaling pathway may result in long-term changes in food intake and body weight. PMID- 11054591 TI - The role of gastrointestinal vagal afferents in the control of food intake: current prospects. AB - Meals are the functional units of food intake in humans and mammals, and physiologic approaches to understanding the controls of meal size have demonstrated that the presence of food in the upper gastrointestinal tract plays a critical role in determining meal size. The vagus nerve is the primary neuroanatomic substrate in the gut-brain axis, transmitting meal-related signals elicited by nutrient contact with the gastrointestinal tract to sites in the central nervous system that mediate ingestive behavior. This article describes progress in examining the role of the vagal gut-brain axis in the negative feedback control of meal size from four perspectives: neuroanatomic, neurophysiologic, molecular, and behavioral. Vagal afferents are strategically localized to be sensitive to meal-related stimuli, and their central projections are organized viscerotopically in the caudal brainstem. Vagal afferents are sensitive to mechanical, chemical, and gut and peptide meal-related stimuli and can integrate multiple such modalities. Meal-elicited gastrointestinal stimuli activate distinct patterns of c-fos neural activation within caudal brainstem sites, where gut vagal afferents terminate. Results of selective chemical and surgical vagal deafferentation studies have refined our understanding of the sites and types of critical gastrointestinal feedback signals in the control of meal size. Recent behavioral, molecular, and neurophysiologic data have demonstrated brainstem sites where centrally acting neuropeptides may modulate the processing of gut vagal afferent meal-related signals to alter feeding. Investigations of the structure and function of splanchnic visceral afferents and enterics and characterization of the integrative capacities of the hindbrain and forebrain components of the gut-brain axis are critical next steps in this analysis. PMID- 11054592 TI - Taste as a factor in the management of nutrition. AB - The sense of taste lies at the interface between the external and internal milieux, at the point at which the animal must decide which chemicals from the environment to incorporate into itself. Accordingly, taste is organized along a neural dimension of nutrients versus toxins, which corresponds to a behavioral dimension of acceptance versus rejection, and to a hedonic dimension of appetitive versus aversive qualities. Reflexive responses, cognitive analyses, and hedonic reactions appear to be managed at different levels of the nervous system. At the first central relay, the nucleus of the solitary tract, somatic reflexes for acceptance or rejection, and autonomic reflexes anticipating digestion are orchestrated. At the second, the parabrachial nucleus of the rodent, associative mechanisms important to the development of conditioned aversions and sodium appetite are manifested. In the thalamic taste relay, gustatory memories associated with non-visceral events may be formed. Primary taste cortex appears to be the site for a cognitive evaluation of gustatory quality and intensity. Finally, a hedonic assessment of the chemical may be made in secondary taste cortex and in the ventral forebrain sites to which it projects. With this assessment comes integration of the gustatory signal with those from other senses, perhaps to create a perception of flavor. Therefore, a sequence that begins with an analysis of the molecular structure of a chemical in the mouth serves to incorporate that gustatory component into an appreciation of flavor, and to participate in the control of motivational processes that guide dietary selection. PMID- 11054593 TI - Elevated and sustained desire for sweet taste in african-americans: a potential factor in the development of obesity. AB - Oral habituation is a relatively long-lasting decrease in oral responsiveness that results from the repeated presentation of a single stimulus. The purpose of this study was to evaluate the degree of habituation to sweet-tasting foods and to determine whether there are differences in the rate of habituation between African Americans and European Americans. These two groups were compared because the prevalence of obesity and obesity-related disorders such as diabetes and hypertension is significantly higher among African Americans than among European Americans. Nine different commercial foods and beverages that differed in sweetness intensity and caloric density served as stimuli. Subjects tasted and rated each food once per minute for a 30-min period on scales related to desire for another taste of the same sample and desire for a different taste. The stimuli and portion size for each of the 30 samples were two candy bars (Ultra Slim-Fast Cocoa Almond Crunch Bar, 1/16 of a bar; Natural Nectar Peanut Butter Granola Bar, 1/16 of a bar), three beverages (Nestea Lemon Flavored Instant Tea with NutraSweet, 5 mL; Welch's Grape Juice, 5 mL; Pink Swimmingo Kool-Aid, 5 mL), two gelatin desserts (Cherry Flavored Jell-O Gelatin, 5 g; Cherry Flavored Jell-O Gelatin with NutraSweet, 5 g), one enteral nutrition drink (Vanilla Ensure Plus, 5 mL), and one pudding (Ultra Slim-Fast Chocolate Pudding, 5 g). Subjects consumed the entire portion of each sample. Habituation occurred for seven of the nine foods as judged by a decrease in the desire for another taste of the same food. The degree of habituation for European Americans and African Americans was similar except for the sweetest food (Cherry Flavored Jell-O Gelatin with NutraSweet), for which African Americans showed no habituation. The degree of habituation in both groups was unrelated to caloric density. Overall, young African Americans had a significantly greater desire for another taste of the same food than did young European Americans for seven of the nine foods, and this desire was strongly correlated with the sweetness intensity for young African Americans but not for young European Americans. Furthermore, young African Americans had a greater desire than young European Americans for a different taste for seven of nine foods. The greater desire for intense sweet tastes may be a factor in the elevated incidence of obesity and diabetes in African Americans. In addition, young African Americans had greater perceived stress in this study than did young European Americans. If African Americans use sweet taste to compensate for feelings of stress, this compensation may also contribute to weight gain. PMID- 11054594 TI - Adiposity signals and the control of energy homeostasis. AB - Recent technologic innovations have enabled probing the workings of individual cells and even molecules. As a result, our knowledge of the biological controls over eating and the regulation of body adiposity is increasing at a rapid pace. We review the evidence that food intake is controlled by separate but interacting groups of molecular signals. One group, termed satiety signals, are proportional to what is being consumed and help to determine meal size. Cholecystokinin is the best known of these, and its premeal administration causes a dose-dependent reduction of meal size. In and of itself, however, cholecystokinin (and other satiety signals) has little impact on body-fat stores. The second group, termed adiposity signals, circulate in proportion to body adiposity and enter the brain, where they interact with satiety signals in the brainstem and hypothalamus. Insulin and leptin are the best known of these adiposity signals, and the administration of either into the brain causes a dose-dependent reduction of both food intake and body weight. Within the brain, parallel but opposing pathways originating in the hypothalamic arcuate nuclei integrate adiposity signals with satiety signals. Those with a net anabolic effect increase food intake and reduce energy expenditure and are represented (among many such signals) by neuropeptide Y; those with a net catabolic effect decrease food intake and energy expenditure and are represented by brain melanocortins. This complex regulatory mechanism allows individuals to adapt their feeding schedule to idiosyncratic environmental constraints, eating whenever it is desirable or possible. Body-weight regulation occurs as adiposity signals alter the efficacy of meal-generated satiety signals. PMID- 11054595 TI - The autonomic nervous system, adipose tissue plasticity, and energy balance. AB - In most mammals, two types of adipose tissue, white and brown, are present. Both are able to store energy in the form of triacylglycerols and to hydrolyze them into free fatty acids and glycerol. Whereas white adipose tissue can provide lipids as substrates for other tissues according to the needs of the organism, brown adipose tissue will use fatty acids for heat production. Over the long term, white fat mass reflects the net balance between energy expenditure and energy intake. Even though these two parameters are highly variable during the life of an individual, most adult subjects remain relatively constant in body weight throughout their lives. This observation suggests that appetite, energy expenditure, and basal metabolic rate are linked. An important characteristic of the adipose tissue is its enormous plasticity for volume and cell-number variations and an apparent change in phenotype between the brown and white adipose tissues. The present review focuses on the cellular mechanisms participating in the plasticity of adipose tissues and their regulation by the autonomic nervous system. There is compelling evidence with regard to the importance of the nervous system in the regulation of adipose tissue mass, either brown or white, by acting on the metabolic pathways and on the plasticity (proliferation, differentiation, transdifferentiation, apoptosis) of these tissues. A better comprehension of the different mechanisms involved in the feedback loop linking the brain and these two types of adipose tissue will lead to a better understanding of the pathophysiology of various disorders including obesity, cachexia, anorexia, and type II diabetes mellitus. PMID- 11054596 TI - Metabolic imprinting on genetically predisposed neural circuits perpetuates obesity. AB - There is an obesity epidemic in the industrialized world that is not simply explained by excess energy intake and decreased energy expenditure. Persistent obesity develops when genetically predisposed individuals are in a chronic state of positive energy balance. Once established, the obese body weight is avidly defended against both over- and underfeeding. Animal studies have shown that lean individuals who are genetically predisposed toward obesity have abnormalities of neural function that prime them to become obese when caloric density of the diet is raised. These neural abnormalities are gradually "corrected" as obesity becomes fully developed, suggesting that obesity is the normal state for such individuals. Thus, defense of the obese body weight may be perpetuated by the formation of new neural circuits involved in energy-homeostasis pathways that are not then easily abolished. Such neural plasticity can occur in both adult life and during nervous-system development. Early pre- and postnatal metabolic conditions (maternal diabetes, obesity, undernutrition) can lead genetically predisposed offspring to become even more obese as adults. This enhanced obesity is associated with altered brain neural circuitry, and these changes can then be passed on to subsequent generations in a feed-forward cycle of ever-increasing body weight. Thus, the metabolic perturbations associated with obesity during both brain development and adult life can produce "metabolic imprinting" on genetically predisposed neural circuits involved in energy homeostasis. Drugs that reduce body weight decrease the defended body weight and alter neural pathways involved in energy homeostasis but have no permanent effect on body weight or neural function in most individuals. Thus, early intervention in mothers, infants, children, and adults may be the only way to prevent the formation of permanent neural connections that promote and perpetuate obesity in genetically predisposed individuals. PMID- 11054597 TI - Neuropeptides and obesity. AB - This review focuses on the expression, content, and release of neuropeptides and on their role in the development of obesity in animal models with single-gene mutations. The balance between neuropeptides that contribute to the control of feeding behavior is profoundly and variously altered in these models, supporting the concept of the existence of several types of obesity. The hypothalamic neuropeptide Y (NPY) and the pro-opiomelanocortin (POMC) systems are the networks most studied in relation to energy intake. Both receive information about the nutritional status and the level of energy storage through insulin and leptin signaling mediated by specific receptors located on POMC and NPY neurons present predominantly in the arcuate nucleus (ARC). When leptin signaling is defective, through a defect in either the receptor (Zucker fa/fa rat, cp/cp rat, and db/db mouse) or in the peptide itself (ob/ob mouse), the NPY system is upregulated as shown by mRNA overexpression and increased peptide release, whereas the content and/or release of some inhibitory peptides (neurotensin, cholecystokinin) are diminished. For the POMC system, there is a complex interaction between the tonic inhibition of food intake exerted by alpha-melanocyte-stimulating hormone (alpha MSH) and the Agouti-related protein at the level of the type 4 melanocortin receptor. The latter peptide is coexpressed with NPY in the ARC. Corticotropin releasing factor (CRF) is the link between food intake and environmental factors. It not only inhibits food intake and prevents weight gain, likely through hypothalamic effects, but also activates the hypothalamo-pituitary axis and therefore contributes to energy storage in adipose tissue. The factors that prod the CRF system toward the hypothalamic or hypothalamo-pituitary axis system remain to be more clearly defined (comodulators, connections between limbic system and ARC, cellular location, and type of receptors, etc. ). The pathways used by all of these neuromodulators include numerous brain areas, but some interest has returned to the classic ones such as the ventromedial and lateral hypothalamic areas because of the recent discovery of some peptides (orexins and melanin-concentrating hormone for the lateral hypothalamus) and receptors (CRF type 2 in the ventromedial hypothalamus). All of these pathways are redundant and function in a coordinated manner and sometimes by the novel expression of a peptide in an unusual area. The importance of such a phenomenon in obesity remains to be determined. Even if single-gene mutations are exceptions in human obesity, the study of genetic animal models of obesity has greatly contributed to the understanding of the regulation of feeding behavior and will allow researchers to develop new drug treatments for obesity that have to be associated with drastic changes in lifestyle (feeding, work habits, and physical activity) for a complete efficiency. PMID- 11054598 TI - Obesity and cortisol. AB - Cortisol in obesity is a much-studied problem. Previous information indicates that cortisol secretion is elevated but that circulatory concentrations are normal or low, suggesting that peripheral disappearance rate is elevated. These studies have usually not taken into account the difference between central and peripheral types of obesity. Recent studies using saliva cortisol have indicated that the problem is complex with both high and low secretion of cortisol, perhaps depending on the status of the function of the hypothalamic-pituitary-adrenal gland axis. A significant background factor seems to be environmental stress. The results also suggest that the pattern of cortisol secretion may be important. Other neuroendocrine pathways are also involved, including the central sympathetic nervous system, the gonadal and growth hormone axes, and the leptin system. In concert, these abnormalities seem to be responsible for the abnormal metabolism often seen in central obesity. Several associated polymorphisms of candidate genes may provide a genetic background. Cortisol conversion to inactive metabolites may be a factor increasing central signals to secretion and may add to the increased secretion of cortisol induced by centrally acting factors. Perinatal factors have been found to be involved in the pathogenesis of obesity and its complications. The mechanism involved is not known, but available information suggests that programming of the hypothalamic-pituitary-adrenal axis may be responsible. PMID- 11054599 TI - Gonadal steroids and energy homeostasis in the leptin era. AB - Gonadal steroids influence food intake and body weight. Although the specific mechanisms underlying these effects are not clear, a consideration of their effects in the context of current models of energy homeostasis may ultimately lead to the identification of these mechanisms. When compared with leptin, the prototypical humoral signal of energy balance, sex steroids share many common properties related to food intake and body weight. Specifically, gonadal steroids circulate in proportion to fat mass and current energy balance, and administration of these compounds influences food intake, energy expenditure, body weight, and body composition. Moreover, both estrogens and androgens modulate central nervous system effectors of energy homeostasis that are targets for the action of leptin, including pathways that contain neuropeptide Y, pro opiomelanocortin, or melanin-concentrating hormone. Sex steroids and leptin also regulate one another's production. Although gonadal steroids, unlike leptin, are clearly not critical to the maintenance of normal energy homeostasis, they do appear to function as physiologic modulators of this process. Identifying the specific central mediators of their effects will contribute to our understanding of their role in energy homeostasis. PMID- 11054600 TI - Obesity and quality of life. AB - The focus of this review is the impact of obesity and weight loss on quality of life. A focus on quality of life broadens the scope of treatment efficacy beyond weight loss and provides a patient-centered perspective. The concept of quality of life is defined, and both general and obesity-specific measures are reviewed. It is clear that obesity confers negative consequences on both the physical and psychosocial aspects of quality of life, especially among the severely obese. The effects of weight loss appear to be favorable, although few studies have examined non-surgical interventions. Future studies would be enhanced by assessing a variety of approaches to weight loss by using both general and obesity-specific measures of quality of life and conducting follow-up studies to assess the effects of weight regain on quality of life. PMID- 11054601 TI - A concise review on the therapeutics of obesity. AB - Drugs to treat obesity can be divided into three groups: those that reduce food intake; those that alter metabolism; and those that increase thermogenesis. Monoamines acting on noradrenergic receptors, serotonin receptors, dopamine receptors, and histamine receptors can reduce food intake. A number of peptides also affect food intake. The noradrenergic drugs phentermine, diethylpropion, mazindol, benzphetamine, and phendimetrazine are approved only for short-term use. Sibutramine, a norepinephrine-serotonin reuptake inhibitor, is approved for long-term use. Orlistat inhibits pancreatic lipase and can block 30% of the triacylglycerol hydrolysis in subjects eating a 30% fat diet. The only thermogenic drug combination that has been tested is ephedrine and caffeine, but this treatment has not been approved by regulatory agencies. In clinical trials other drugs that may modulate peptide-feeding systems are being developed. PMID- 11054602 TI - Influence of topiramate in the regulation of energy balance. AB - Topiramate (TPM) is a novel neurotherapeutic agent currently indicated for the treatment of epilepsy and undergoing development for other central nervous system indications including neuropathic pain, bipolar disorder, and migraine prophylaxis. TPM is synthesized from D-fructose and contains a sulfamate moiety that is essential for its pharmacologic activity. TPM has been observed to significantly reduce body weight in patients treated for seizure, which has prompted the realization of preclinical studies to characterize the effects of TPM in the regulation of energy balance. Studies carried out in various strains of rats have provided good evidence for the ability of TPM to blunt energy deposition. Body composition analyses from rat trials have demonstrated that TPM inhibits fat deposition while reducing the activity of lipoprotein lipase (LPL) in various white adipose tissue depots. High doses of TPM (likely above the therapeutic dose range) have also been observed to reduce protein gain without catabolic effects. Although TPM cannot be described as a potent anorectic agent, it seems to have the ability to reduce food intake; significant reductions in food intake have been observed in female obese (fa/fa) Zucker rats and in female Wistar rats. TPM can also reduce energy deposition in the absence of alterations in food intake. This effect has been clearly emphasized in female lean (Fa/?) Zucker rats. In female Sprague-Dawley rats, TPM also increased energy expenditure and it has been observed to increase LPL activity in brown adipose tissue, which could indicate that TPM has the ability to enhance regulatory thermogenesis. In addition, TPM stimulates LPL activity in skeletal muscles, further emphasizing its potential to promote substrate oxidation. The mechanisms whereby TPM affects the regulation of energy balance have yet to be understood. TPM represents an antiepileptic drug (AED) with complex biochemical/pharmacologic actions. Its negative effects on energy deposition cannot be readily predicted from these actions, as AEDs are generally expected to stimulate body weight gain. Recent data, obtained from investigations aimed at assessing the effects of TPM on neuropeptidergic systems involved in the regulation of energy balance, have failed to demonstrate any significant effects of TPM on the neuropeptide Y and proopiomelanocortin systems. In conclusion, it is clear that TPM can reduce fat deposition by either reducing food intake or stimulating energy expenditure. The mechanisms whereby an AED such as TPM controls food intake and energy expenditure remains to be delineated. Copyright1999 ASCRS and ESCRS PMID- 11054603 TI - Effect of topiramate on body weight and body composition of osborne-mendel rats fed a high-fat diet: alterations in hormones, neuropeptide, and uncoupling protein mRNAs. AB - The effects of topiramate on food intake and body composition were investigated in rats fed a high-fat diet and compared with rats that were pair fed or treated with D-fenfluramine. Topiramate (40 mg. kg. d for 80 d) reduced body-weight gain in a manner similar to that of pair-fed rats and D-fenfluramine-treated rats. The reduction in body fat accounted for all the weight reduction after topiramate treatment but not after pair feeding or D-fenfluramine treatment. Topiramate reduced food intake acutely and increased metabolic rate. There were also significant reductions in leptin, insulin, and corticosterone. In the hypothalamus, topiramate increased mRNA for neuropeptide Y, reduced mRNA for neuropeptide-Y Y1 and Y5 receptors, corticotropin-releasing hormone (CRH), and type II glucocorticoid receptors but had no effect on mRNA levels for the short or long form of the leptin receptor. In peripheral tissues, topiramate reduced leptin mRNA in adipose tissue, had no effect on uncoupling protein 1 mRNA in brown adipose tissue but had tissue-selective effects on uncoupling proteins 2 and 3 mRNA levels in white and brown adipose tissues and muscle. In conclusion, topiramate is an effective inhibitor of weight gain in rats on a high-fat diet, but the mechanism through which the change in energy balance is achieved is unclear. PMID- 11054604 TI - The future of obesity research. AB - As endocrinologists, we view better treatment as the goal of obesity research. The ideal obesity treatment would reduce body fat substantially, with preferential loss from the visceral compartment, and preserve lean tissue with a minimum of side effects. Obesity has been recognized as a chronic disease since 1985. Chronic diseases recognized before obesity may predict the future of obesity research. Initial treatments of chronic diseases commonly arise from empirical observations. These observations often stimulate basic research into the physiologic mechanisms responsible. Such cross-fertilization between the clinic and basic science is desirable and expected. As more is learned about the physiology of obesity, treatments can be expected to use more downstream mechanisms with less unwanted side effects, the reliance on surgical treatments can be expected to decline, and molecular approaches are likely to play an increasingly important role. With a better physiologic understanding of obesity, advanced clinical endpoints will become more important and molecular approaches are likely to play a more important role in discovery and treatment. Due to the availability of molecular approaches, obesity treatment is expected to advance faster than chronic disease research of the past. PMID- 11054605 TI - The anorexia of aging. PMID- 11054606 TI - Anorexia of infection: current prospects. AB - The anorexia of infection is part of the host's acute phase response (APR). Despite being beneficial in the beginning, long lasting anorexia delays recovery and is ultimately deleterious. Microbial products such as bacterial cell wall compounds (e.g., lipopolysaccharides and peptidoglycans), microbial nucleic acids (e. g., bacterial DNA and viral double-stranded RNA), and viral glycoproteins trigger the APR and presumably also the anorexia during infections. Microbial products stimulate the production of proinflammatory cytokines (e.g., interleukins [ILs], tumor necrosis factor-alpha, interferons), which serve as endogenous mediators. Several microbial products and cytokines reduce food intake after parenteral administration, suggesting a role of these substances in the anorexia during infection. Microbial products are mainly released and cytokines are produced in the periphery during most infections; they might inhibit feeding through neural and humoral pathways activated by their peripheral actions. Activation of peripheral afferents by locally produced cytokines is involved in several cytokine effects, but is not crucial for the anorectic effect of microbial products and IL-1beta. Cytokines increase leptin expression in the adipose tissue, and leptin may contribute to, but is also not essential for, the anorectic effects of microbial products and cytokines. In addition, a direct action of cytokines and microbial products on the central nervous system (CNS) is presumably involved in the anorexia during infection. Cytokines can reach CNS receptors through circumventricular organs and through active or passive transport mechanisms or they can act through receptors on endothelial cells of the brain vasculature and stimulate the release of subsequent mediators such as eicosanoids. De novo CNS cytokine synthesis occurs in response to peripheral infections, but its role in the accompanying anorexia is still open to discussion. Central mediators of the anorexia during infection appear to be neurochemicals involved in the normal control of feeding, such as serotonin, dopamine, histamine, corticotropin releasing factor, neuropeptide Y, and alpha melanocyte-stimulating hormone. Reciprocal, synergistic, and antagonistic interactions between various pleiotropic cytokines, and between cytokines and neurochemicals, form a complex network that mediates the anorexia during infection. Current knowledge on the mechanisms involved suggests some therapeutic options for treatment. Substances that block common key steps in cytokine synthesis or cytokine action, or inhibitors of eicosanoid synthesis, may hold more promise than attempts to antagonize specific cytokines. To target the neurochemical mediation of the anorexia during infection may be even more efficient. Future research should address these neurochemical mechanisms and the cytokine actions at the blood-brain barrier. Further unanswered questions concern the modulation of the anorexia during infection by gender and nutritional state. PMID- 11054607 TI - Cachexia-anorexia workshop. Vancouver, British Columbia, Canada. November 7-9, 1997. PMID- 11054608 TI - Central nervous system mechanisms contributing to the cachexia-anorexia syndrome. AB - The cachexia-anorexia syndrome occurs in chronic pathophysiologic processes including cancer, infection with human immunodeficiency virus, bacterial and parasitic diseases, inflammatory bowel disease, liver disease, obstructive pulmonary disease, cardiovascular disease, and rheumatoid arthritis. Cachexia makes an organism susceptible to secondary pathologies and can result in death. Cachexia-anorexia may result from pain, depression or anxiety, hypogeusia and hyposmia, taste and food aversions, chronic nausea, vomiting, early satiety, malfunction of the gastrointestinal system (delayed digestion, malabsorption, gastric stasis and associated delayed emptying, and/or atrophic changes of the mucosa), metabolic shifts, cytokine action, production of substances by tumor cells, and/or iatrogenic causes such as chemotherapy and radiotherapy. The cachexia-anorexia syndrome also involves metabolic and immune changes (mediated by either the pathophysiologic process, i.e., tumor, or host-derived chemical factors, e.g., peptides, neurotransmitters, cytokines, and lipid-mobilizing factors) and is associated with hypertriacylglycerolemia, lipolysis, and acceleration of protein turnover. These changes result in the loss of fat mass and body protein. Increased resting energy expenditure in weight-losing cachectic patients can occur despite the reduced dietary intake, indicating a systemic dysregulation of host metabolism. During cachexia, the organism is maintained in a constant negative energy balance. This can rarely be explained by the actual energy and substrate demands by tumors in patients with cancer. Overall, the cachectic profile is significantly different than that observed during starvation. Cachexia may result not only from anorexia and a decreased caloric intake but also from malabsorption and losses from the body (ulcers, hemorrhage, effusions). In any case, the major deficit of a cachectic organism is a negative energy balance. Cytokines are proposed to participate in the development and/or progression of cachexia-anorexia; interleukin-1, interleukin-6 (and its subfamily members such as ciliary neurotrophic factor and leukemia inhibitory factor), interferon-gamma, tumor necrosis factor-alpha, and brain-derived neurotrophic factor have been associated with various cachectic conditions. Controversy has focused on the requirement of increased cytokine concentrations in the circulation or other body fluids (e.g., cerebrospinal fluid) to demonstrate cytokine involvement in cachexia-anorexia. Cytokines, however, also act in paracrine, autocrine, and intracrine manners, activities that cannot be detected in the circulation. In fact, paracrine interactions represent a predominant cytokine mode of action within organs, including the brain. Data show that cytokines may be involved in cachectic-anorectic processes by being produced and by acting locally in specific brain regions. Brain synthesis of cytokines has been shown in peripheral models of cancer, peripheral inflammation, and during peripheral cytokine administration; these data support a role for brain cytokines as mediators of neurologic and neuropsychiatric manifestations of disease and in the brain-to-peripheral communication (e.g., through the autonomic nervous system). Brain mechanisms that merit significant attention in the cachexia anorexia syndrome are those that result from interactions among cytokines, peptides/neuropeptides, and neurotransmitters. These interactions could result in additive, synergistic, or antagonistic activities and can involve modifications of transducing molecules and intracellular mediators. Thus, the data show that the cachexia-anorexia syndrome is multifactorial, and understanding the interactions between peripheral and brain mechanisms is pivotal to characterizing the underlying integrative pathophysiology of this disorder. PMID- 11054609 TI - Metabolic abnormalities in cachexia and anorexia. AB - An increased glucose requirement by many solid tumors produces an increased metabolic demand on the liver, resulting in an increased energy expenditure. In addition, several cytokines and tumor catabolic products have been suggested as being responsible for the depletion of adipose tissue and skeletal-muscle mass in cachexia. A sulphated glycoprotein of molecular mass 24 kDa, produced by cachexia inducing tumors and present in the urine of cancer patients actively losing weight, has been shown to be capable of inducing direct muscle catabolism in vitro and a state of cachexia in vivo, with specific loss of the non-fat carcass mass. In vitro studies have shown the bioactivity of this proteolysis-inducing factor to be attenuated by the polyunsaturated fatty acid, eicosapentaenoic acid. Preliminary clinical studies have shown that eicosapentaenoic acid stabilizes body weight and protein and fat reserves in patients with pancreatic carcinoma. Further trials are required to confirm the efficacy of eicosapentaenoic acid and to determine the anticachectic activity in other types of cancer. PMID- 11054610 TI - Regulation of skeletal-muscle-protein turnover in cancer-associated cachexia. AB - Cancer is frequently associated with anorexia, weight loss, negative nitrogen balance, and skeletal-muscle wasting. Depletion of skeletal-muscle mass is critical to overall survival of the patient, can prolong rehabilitation to normal function after recovery, and decreases quality of life in a palliative-care setting. The biochemical and physiologic bases of cancer-associated muscle wasting have been most fully investigated in animal models. These studies provide evidence for suppressed protein synthesis and activated proteolysis in cancer associated muscle wasting and indicate a need for both anabolic and anticatabolic therapies. Several humoral factors of host or tumor origin are implicated in altered muscle-protein metabolism, including cytokines, metabolites of arachidonic acid, and a proteolysis-inducing glycoprotein; their interrelationships are less well characterized. Several catabolic mediators may share common downstream mechanisms because they ultimately activate the ATP-, ubiquitin-, and proteasome-dependent intracellular proteolytic system. Although important gaps in our current understanding remain, data available from animal studies can be used as a basis to develop relevant studies in human subjects. PMID- 11054612 TI - Macronutrients and mental performance. AB - There is currently intense interest in the effects of macronutrients on psychological states, mental performance, and well-being. A strong theoretical perspective has guided work on carbohydrates and their relation to brain serotoninergic function with concomitant effects on performance. The clearest and most reliable effects have been observed for the beneficial action of glucose on cognitive performance, supported by investigations of hypoglycemia, which is associated with general impairment of cognitive performance. The effects of complex carbohydrates are less distinct and change with time of day; e.g., carbohydrate at breakfast tends to improve morning performance. However, these studies are rarely decisive. Far fewer experiments have been performed on protein and fat, and it is difficult to draw any firm conclusions. Macronutrients are seldom given alone, proportions of protein and fat differ greatly between studies, and comparisons are frequently performed with no food at all. Food intake may mitigate the effects of low doses but not of high doses of alcohol on performance. Effects of macronutrients on cognitive performance may be dependent on their effects on glucose metabolism, metabolic activation, or serotonin. Other factors that modify effects include time of day, circadian rhythms, type of task, habitual diet, and vulnerability of the population. PMID- 11054613 TI - Dofetilide: a new pure class III antiarrhythmic agent. AB - BACKGROUND: Although there are a variety of antiarrhythmic agents used for the treatment of atrial fibrillation of flutter, each drug has drawbacks, and room exists for new pharmacologic agents. Dofetilide, a pure class III agent, has recently been approved by the Food and Drug Administration for therapy of these arrhythmias and is reviewed. METHODS: Data for dofetilide, published in full or in abstract form, were reviewed, concentrating on the properties related to its efficacy for the therapy of supraventricular arrhythmias. RESULTS: Results from animal and human studies indicate that dofetilide, a renally excreted drug, has pure class III properties related to blockade of the delayed rectifier potassium current. It is effective for the therapy of atrial arrhythmias, particularly atrial fibrillation and flutter, and has no demonstrable negative inotropic effect. Despite an incidence of torsades de pointes of approximately 2% in patients with impaired ventricular function, dofetilide exhibited no association with an increased mortality rate when studied in a large series of patients with a reduced ejection fraction. CONCLUSIONS: Dofetilide's electrophysiologic and clinical profiles suggest that it will be safe and clinically useful for the termination and prevention of atrial fibrillation or flutter, even in patients with impaired ventricular function. PMID- 11054614 TI - Multimodality reperfusion therapy for acute myocardial infarction. AB - With the strong and direct relation between early reperfusion in acute myocardial infarction (AMI) and improved clinical outcomes, attention has focused on new means of improving rates of reperfusion and accelerating every stage of AMI evaluation and management, from the onset of symptoms of myocardial infarction to the achievement of reperfusion. Critical pathways to streamline the evaluation and management of AMI have cut minutes and even hours off in-hospital treatment times for patients with AMI; public health initiatives focus on educational efforts to shorten time to hospital arrival. The latest advance in fibrinolytic therapy is the availability of bolus fibrinolytic agents with safety and efficacy in large phase III trials comparable to accelerated intravenous infusion regimens. Faster and simpler fibrinolytic regimens may shorten door-to-needle time, reduce medication errors, and facilitate prehospital thrombolysis. Bolus fibrinolytic agents are being evaluated for use in combination with other interventions to open occluded coronary arteries, including acute percutaneous coronary intervention, the glycoprotein IIb/IIIa platelet inhibitors, or both. The goal of this "multimodality" approach to AMI management is to minimize time to reperfusion and maximize the percentage of patients who achieve complete arterial patency and myocardial perfusion without bleeding complications. PMID- 11054615 TI - Evaluation of early postoperative results after bicaval versus standard cardiac transplantation and review of the literature. AB - OBJECTIVE: Previous studies have been inconsistent in defining a clinical benefit to the bicaval cardiac transplantation technique relative to the standard technique, and many major centers have not adopted this newer approach. The purpose of this study was to determine whether clinically significant benefits support utilization of the bicaval technique. METHODS: Sixty-eight consecutive adult patients undergoing a standard cardiac transplant were compared with 75 consecutive patients who underwent the bicaval technique during the period from 1991 to 1999. Etiology, recipient sex, recipient age, donor age, and pulmonary vascular resistance were similar between the two groups. RESULTS: Cardiac index at 24 hours after operation was increased for the bicaval group relative to the standard group (3.15 +/- 0.7 vs 2.7 +/- 0.5 L/min/m(2), P <. 05). Inotropic requirements were significantly less, and there was significantly less tricuspid regurgitation in the bicaval group relative to the standard group. In addition, the bicaval group more frequently had a nonpaced normal sinus rhythm at 24 hours after operation (73.9% vs 50.7% [standard group], P =.025) and had fewer postoperative arrhythmias (29.3% vs 47.7% [standard group], P <.01). Finally, although mortality was similar for the two groups, length of postoperative hospitalization was longer for the standard group relative to the bicaval group (12.1 +/- 11 vs 20.4 +/- 12 days, P <. 001). Review of the literature identified reduced tricuspid regurgitation and improved rhythm as consistent benefits of the bicaval technique. CONCLUSION: This review demonstrates a clinical benefit during the early postoperative period with bicaval cardiac transplantation (relative to standard) and encourages further utilization of this technique. PMID- 11054616 TI - A direct comparison of tirofiban and abciximab during percutaneous coronary revascularization and stent placement: rationale and design of the TARGET study. AB - BACKGROUND: Trials testing intravenous platelet glycoprotein IIb/IIIa antagonists in the setting of percutaneous coronary revascularization and empirically during acute coronary syndromes have consistently demonstrated a reduction in ischemic events. These trials, however, have varied regarding patient population, type, duration and timing of IIb/IIIa therapy, adjunct therapies, and methods for collection and adjudication of end points. All trials were placebo-controlled, and none involved a direct comparison of IIb/IIIa inhibitors. Whether these agents produce a similar clinical outcome in the contemporary practice of coronary interventions is uncertain. METHODS AND RESULTS: To evaluate the efficacy of tirofiban in patients undergoing percutaneous revascularization with stent placement, a randomized, multicenter, double-blind, double-dummy, abciximab controlled study is currently underway. All patients will receive preprocedural clopidogrel, weight-adjusted heparin, and aspirin. In 18 countries, 4750 patients undergoing nonemergency percutaneous coronary revascularization will be studied. The primary end point will be the composite 30-day occurrence of death, myocardial infarction, or urgent target vessel revascularization. Secondary end points will include 6-month death, myocardial infarction, or any myocardial revascularization and 1-year death. CONCLUSION: This is the first large-scale, head-to-head comparison of 2 established IIb/IIIa inhibitors in interventional cardiology. Enrollment is expected to be complete by mid-2000. PMID- 11054617 TI - Valsartan in acute myocardial infarction trial (VALIANT): rationale and design. AB - BACKGROUND: Survivors of acute myocardial infarction (MI) complicated by heart failure and/or resulting in left ventricular dysfunction are at heightened risk for subsequent death and major nonfatal cardiovascular events. Inhibition of the renin-angiotensin system with an angiotensin-converting enzyme inhibitor has consistently been demonstrated to result in reductions in these risks by approximately 20%. The development of angiotensin II receptor blockers offers a new, more specific, and theoretically more complete pharmacologic mode to inhibit the adverse influence of angiotensin II. METHODS: Valsartan in Acute Myocardial Infarction (VALIANT) is a multicenter, double-blind, randomized, active controlled parallel group study comparing the efficacy and safety of long-term treatment with valsartan, captopril, and their combination in high-risk patients after MI. The trial is designed with 3 arms, giving equal statistical consideration to survival comparisons of captopril versus the angiotensin II receptor blocker valsartan, as well as the combination of captopril plus valsartan, compared with a proven effective dose of captopril. This 14,500 patient trial is designed with an 86% power to detect a 15% reduction in mortality rate with either use of valsartan compared with captopril. The trial encourages optimal individualization of other proven therapies in acute and chronic infarction, and the international patient body ensures good representation of multiple practice patterns. CONCLUSION: VALIANT is a large international investigative effort that will evaluate the role of valsartan in the management of patients with MI associated with heart failure and/or left ventricular dysfunction. The use of a proven dose of captopril and the comparator arms with valsartan alone or in combination with captopril provides a unique test of whether the angiotensin II receptor blocker can make an additional improvement in clinical outcomes beyond angiotensin-converting enzyme inhibitors. PMID- 11054618 TI - The defibrillator in acute myocardial infarction trial (DINAMIT): study protocol. AB - BACKGROUND: The implantable cardioverter/defibrillator (ICD) has been shown to be superior to antiarrhythmic drug therapy for the secondary prevention of sudden cardiac death. Its role in the primary prevention of sudden death after myocardial infarction is unknown. Methods and Results The Defibrillator in Acute Myocardial Infarction Trial (DINAMIT) is a randomized, open-label, parallel-group comparison of ICD therapy versus no ICD therapy in selected survivors of acute myocardial infarction. It will test the hypothesis that reduction of sudden arrhythmogenic death by means of the ICD will result in reduction of overall mortality rates in patients at high risk after acute myocardial infarction. Accordingly, this international multicenter study aims to enroll patients shortly after their infarction (day 6 to day 40) who have reduced left ventricular function (left ventricular ejection fraction /=12 atm) until the achievement of angiographic success (<20% residual stenosis) with a prespecified very high inflation pressure (VHP) strategy of 20 atm without intermediate inflations. METHODS AND RESULTS: We conducted a parallel-group, nonrandomized study to evaluate the short- and long term results in 136 consecutive eligible patients who underwent successful single Palmaz-Schatz stent implantation in vessels >/=3 mm. Major adverse cardiac events (MACE), that is, death, myocardial infarction, and target lesion revascularization (TLR), were monitored for a minimum of 6 months. No significant differences were observed between the two strategies in terms of final minimal lumen diameter (HP, 3.0 +/- 0.5 vs VHP, 3. 1 +/- 0.5 mm) and acute gain (HP, 2.1 +/- 0.7 vs VHP, 2.2 +/- 0.6). The overall rate of subacute stent thrombosis was 0.7%. During a 405 +/- 148-day follow-up, 21 (28.8%) patients in the VHP group and 6 (9. 5%) in the HP group (P =.005) had MACE, with a TLR rate of 27.4% versus 7.9% (P =.009), respectively. By multivariate analysis, the use of VHP increased the odds of long-term MACE by a factor of 3.48 (P =.009). Among patients undergoing TLR, those treated with VHP had a greater lumen loss (HP, 1.83 +/- 0.57 vs VHP, 2.15 +/- 0.36 mm, P =.02) and a more frequent pattern of diffuse restenosis (71% vs 16%, P =.06). CONCLUSIONS: In our study, the two strategies had similar acute and short-term results, but VHP was associated with a poorer long-term outcome. These data provide a rationale for a less aggressive strategy for stent deployment by optimizing rather than attempting to maximize inflation pressure and stent expansion. PMID- 11054630 TI - Preconditioning during percutaneous transluminal coronary angioplasty by endogenous and exogenous adenosine. AB - BACKGROUND: The purpose of this study was to assess whether pharmacologic preconditioning by exogenous or endogenous adenosine prevents the deterioration of hemodynamic function and left ventricular performance during percutaneous transluminal coronary angioplasty (PTCA). Ischemic preconditioning renders the heart more resistant to subsequent ischemia. Adenosine plays a key role in its pathogenesis. Coronary angioplasty is a suitable model for the induction of myocardial ischemia. METHODS AND RESULTS: We investigated 30 patients receiving PTCA of the left anterior descending coronary. Patients were randomly allocated to either dipyridamole, leading to the liberation of endogenous adenosine (0.5 mg/kg body weight, intracoronary), exogenous adenosine (20 mg intracoronary), or an equal amount of saline. Chest pain, tolerated inflation time, and ST-segment shift were registered. Left ventricular hemodynamics, isovolumetric phase indexes, indexes of volume, ejection fraction, and indexes of diastolic dysfunction were analyzed. Patients receiving endogenous or exogenous adenosine tolerated longer balloon inflation times (dipyridamole, 208 +/- 23 seconds; adenosine, 188 +/- 41 seconds; control, 153 +/- 36 seconds; P <.05). Deterioration of left ventricular ejection fraction was less severe after adenosine (72% +/- 5% before PTCA vs 64% +/- 6% during angioplasty; P =.11) and could be prevented by intracoronary dipyridamole (69% +/- 12% before PTCA vs 68% +/- 11% after PTCA; P <. 01) compared with the control group (71% +/- 7% before PTCA vs 60% +/- 7% during angioplasty). CONCLUSIONS: Intracoronary application of exogenous adenosine and liberation of endogenous adenosine increase the tolerance to ischemia and prevent deterioration of left ventricular function during ischemia. These findings can be attributed to ischemic preconditioning. However, endogenous adenosine exceeds the protective effects of exogenous adenosine. PMID- 11054631 TI - Immediate stent implantation versus conventional techniques for the treatment of abrupt vessel closure or symptomatic dissections after coronary balloon angioplasty. AB - BACKGROUND: Coronary stenting was initially designed to treat a bailout scenario. Prospective randomized trials comparing stent implantation with standard techniques, including emergency coronary artery bypass grafting, are lacking. The aim of this trial was to test the superiority of immediate stent implantation compared with standard techniques for the treatment of abrupt or threatening closure after coronary balloon angioplasty. METHODS: In a prospective trial, 100 patients with abrupt vessel closure or symptomatic dissections causing objective signs of ischemia were randomly assigned to treatment with immediate placement of stents (n = 51) versus standard techniques such as prolonged dilatation or emergency bypass surgery (n = 49). The primary end point was the achievement of successful stabilization not requiring crossover to the other study group. Secondary end points included event-free survival and restenosis. RESULTS: Successful stabilization was achieved in 94% of patients in the stent group compared with 78% of patients in the standard treatment group (P =.038). Two patients died in each group, and there was a trend toward a higher incidence of myocardial infarction (16% vs 8%; P =.163) and a significantly increased creatine phosphokinase level (245 IU/L [95% confidence interval, 217-265 IU/L] vs 179 IU/L [confidence interval 140-212 IU/L]; P =.0002) in the standard treatment group. Event-free survival after 250 days was 72% in the stent group compared with 29% in the standard treatment group (P =.001). The angiographic restenosis rate was 30% in the stent group versus 59% in the standard treatment group (P =.01). CONCLUSIONS: Immediate stenting, if technically feasible, shows superior short- and long-term results compared with standard treatment options. PMID- 11054632 TI - Cardiomegaly as a possible cause of lung dysfunction in patients with heart failure. AB - BACKGROUND: Our hypothesis is that an enlarged heart may compete for space with the lungs, causing a restrictive pattern that is often seen in patients with chronic heart failure. METHODS: Eighty patients with stable congestive heart failure in New York Heart Association classes II and III participated in the study. We measured cardiothoracic index (chest radiography), FEV1, vital capacity, alveolar volume, lung diffusion capacity for carbon monoxide (DLCO), and its 2 subcomponents alveolar-capillary membrane diffusion (DM), and pulmonary capillary blood volume. RESULTS: Reliable measurements were obtained in 72 of 80 participants enrolled. Cardiothoracic index averaged 57% +/- 7%. FEV1, vital capacity, alveolar volume, DLCO, and DM were inversely related to the cardiothoracic index (r = -0.514, -0.557, -0.522, -0.475, and -0.480, respectively). However, the relations of DLCO and DM with the cardiothoracic index were lost when DLCO and DM were adjusted for alveolar volume. A significant correlation (P < .01) was found between alveolar volume and vital capacity, FEV1, and DLCO (r = 0.799, 0.705, and 0.614, respectively). At multivariate analysis, cardiothoracic index, FEV1, and pulmonary capillary blood volume were independent predictors of DLCO, whereas alveolar volume, FEV1, and left ventricular ejection fraction were independent predictors of DM. CONCLUSIONS: Cardiac enlargement in chronic heart failure appears to be involved in causing restrictive lung pattern and a reduced alveolar volume that disturbs carbon monoxide diffusion. PMID- 11054633 TI - Increased levels of clusterin (SGP-2) mRNA and protein accompany rat ventral prostate involution following finasteride treatment. AB - Finasteride is a well-known inhibitor of the prostatic enzyme 5 alpha-reductase type 2 which prevents conversion of testosterone into 5 alpha dihydrotestosterone, the active intraprostatic androgen, which causes prostate involution through a combination of cell atrophy and cell death. The drug is widely used to improve symptoms of benign prostatic hyperplasia in man. Clusterin, a glycoprotein which is generally up-regulated under conditions inducing cell atrophy or organ involution, is produced at a high level in the regressing rat ventral prostate following androgen ablation. According to several authors, clusterin does not respond to finasteride treatment, suggesting a different action of testosterone and 5 alpha-dihydrotestosterone. We show here that, under our conditions, finasteride was capable of inducing production of both clusterin mRNA and protein in the rat ventral prostate. In fact, by using different and converging techniques, such as Northern hybridization, in situ hybridization histochemistry and immunohistochemistry, we were able to show a strong induction of the clusterin gene in the epithelial cell population of the gland. The response to finasteride, which was similar to that seen with castration, occurred with a delay of a few days. In situ and immunohistochemistry experiments indicated that both orchidectomy and finasteride administration resulted in increased transition of the epithelial cells from the columnar to the cuboidal (atrophic) shape, and this was accompanied by an increased intensity of the signal for clusterin. Thus, it appears that induction of clusterin is part of the molecular process leading to prostate involution caused by either orchidectomy or finasteride administration. PMID- 11054634 TI - Post-surgical recovery and time-of-day mask potentiated responses of ACTH to repeated moderate hemorrhage in conscious rats. AB - We determined how changes in the responsiveness of the hypothalamo-pituitary adrenal (HPA) system that accompany experimentation affect facilitation of HPA responses to hemorrhage. Hemorrhage (10 ml/kg over 3 min) was performed in conscious, chronically prepared rats. Blood was sampled over 1 h followed by reinfusion of shed blood. Hemorrhage was performed either once or twice separated by 24 h in different groups of animals. To test the effect of the circadian variation in responsiveness, rats were hemorrhaged on days 4 and 5 after surgery either in the morning (AM) or in the afternoon (PM). The response of ACTH to hemorrhage on day 4 was greater in the PM than in the AM (P<0.01). The ACTH response to the second hemorrhage on day 5 was greater than that to hemorrhage on day 4 only in the AM group (P<0.01). Thus, facilitation of ACTH responses by prior hemorrhage was evident only in the AM. To determine the effects of surgical recovery, additional experiments were done in the AM either early (days 3 and 4) or later (days 6 and 7) after surgery. In these experiments, hemorrhage was performed in all rats on days 4 and 7 and either hemorrhage or blood sampling alone was performed on day 3 and 6. ACTH did not increase in rats with sampling and no hemorrhage. ACTH increased more after an initial hemorrhage on day 3 than on day 6 (P<0.01). ACTH response to hemorrhage on day 4 was greater when preceded by hemorrhage vs sampling on day 3 (P<0.01). ACTH response to hemorrhage in rats bled twice did not differ on day 3 and day 4. On day 7, the response of ACTH in rats that had hemorrhage on day 6 was greater than both their own response on day 6 and the response of a control group with sampling on day 6 (P<0.01). These results demonstrate potentiation of ACTH responses to hemorrhage by an earlier similar hemorrhage, but clearly indicate that enhanced sensitivity of the HPA to hemorrhage either by circadian factors or by surgery can mask this effect. PMID- 11054635 TI - Differences between the silencing-related properties of the extreme carboxyl terminal regions of thyroid hormone receptors alpha 1 and beta 1. AB - Human thyroid hormone receptor (TR) is encoded by two distinct genes, TR alpha and TR beta. TR heterodimerizes with retinoid X receptor (RXR) and binds efficiently to the thyroid hormone (T(3)) response element (TRE) of target genes. In the absence of T(3), unliganded TR suppresses the basal promoter activity of positively regulated genes (silencing). Silencing mediator for retinoid and thyroid hormone receptors (SMRT) and nuclear receptor co-repressor (N-CoR) interact with unliganded TR and function as corepressor proteins. Previously, we found beta F451X with carboxyl (C)-terminal 11-amino acid deletion had stronger silencing potency than wild-type TR beta 1 and beta E449X with C-terminal 13 amino acid deletion on a subset of TREs. In the present study, to assess the isoform-specific effects of the C-terminal truncations on TR silencing, we constructed two mutant TR alpha 1s (alpha F397X and alpha E395X) with the same respective C-terminal truncations as beta F451X and beta E449X and analysed their silencing activities. Unlike beta F451X and beta E449X, alpha F397X and alpha E395X showed similarly stronger silencing potency than wild-type TR alpha 1. We further studied the abilities of wild-type and the mutant TR beta 1s and alpha 1s on RXR and co-repressor binding by a two-hybrid interference assay. beta F451X had significantly stronger abilities to bind to RXR and SMRT than did wild-type TR beta 1 and beta E449X. In contrast, wild-type TR alpha 1, alpha F397X and alpha E395X showed similar abilities to bind to RXR and SMRT. beta E449X and alpha E395X, which have identical C-terminal truncation, showed less ability to bind to N-CoR than did wild-type TR beta 1 and beta F451X and wild-type TR alpha 1 and alpha F397X respectively. These results indicate that an identical C terminal truncation gives rise to different effects on TR beta 1 and alpha1 with respect to silencing potency, RXR binding and SMRT binding. The difference in the silencing potency among wild-type TR beta 1, beta F451X and beta E449X correlated well with the difference in the ability to bind co-repressor SMRT. PMID- 11054636 TI - Beneficial effects of retinoic acid on extracellular matrix degradation and attachment behaviour in follicular thyroid carcinoma cell lines. AB - The prognosis of patients with metastasised follicular thyroid carcinoma (FTC) is limited, necessitating the search for new treatment options. Beneficial effects of retinoids have been suggested in thyroid cancer and the present study was performed to investigate the effects of retinoic acid (RA) on important determinants of metastatic behaviour in FTC: the disengagement of tumour cells from the primary tumour and the degradation of extracellular matrix, focusing on the role of the plasmin activation system and the integrin and E-cadherin families of attachment molecules. Three FTC cell lines were studied: FTC-133, derived from the primary tumour; and FTC-236 and FTC-238, derived from metastases. FTC cell lines were cultured with 0.1, 1 and 10 microM 13-cis-RA or with the solvent DMSO for 1 and 5 days. Extracellular matrix degradation by these cell lines was studied by assessing the 48-h release of radioactivity from (35)S methionine labelled extracellular matrix proteins synthesised by the MC3T3 cell line coated onto plastic. The involvement of constituents of the plasmin activation system was investigated by semi-quantitative RT-PCR and zymography. Attachment to extracellular matrix was studied by determining the number of adhering FTC cells to extracellular matrix coated onto plastic, 3 h after seeding. The involvement of attachment molecules was studied by RT-PCR with primers for integrin subclasses and E-cadherin and immunofluorescence for E cadherin. Five days culturing with 10 microM RA reduced the degradation of extracellular matrix significantly in all cell lines: FTC-133 by 35%, FTC-236 by 74% and FTC-238 by 31%. Zymography revealed diminished activity of urokinase type plasminogen activator (uPA) in FTC-236 and FTC-238, but not in FTC-133 cultured with RA. mRNA expression of the uPA receptor was diminished in FTC-236. In the attachment assay, 10 microM RA for 5 days increased the number of adherent cells to extracellular matrix significantly by 91% in FTC-133, 64% in FTC-236 and 87% in FTC-238. No effects of RA on integrin or E-cadherin mRNA expression were observed. Immunofluorescence, however, revealed enhanced organisation of E cadherin along the cell membrane by RA treatment. In conclusion, the present study demonstrates beneficial effects of RA on important determinants of metastatic behaviour in FTC cell lines, e.g. decreased degradation of extracellular matrix which may in part be explained by effects on the plasmin activation system and enhanced attachment to extracellular matrix. These findings may add to the explanations for beneficial effects of retinoids in thyroid cancer. PMID- 11054637 TI - Delayed metabolism of human brain natriuretic peptide reflects resistance to neutral endopeptidase. AB - Metabolism of natriuretic peptides is regulated by two degradative pathways: uptake by the clearance receptor (natriuretic peptide receptor C--NPR-C) and hydrolysis by neutral endopeptidase (NEP). Affinity studies favour a dominant role of NPR-C in hormone degradation in several species but do not account for the efficacy of NEP inhibitors in vivo, nor the uniquely prolonged half life (t((1/2))) of human brain natriuretic peptide (hBNP). Postulating that (1) delayed metabolism of hBNP reflects resistance to NEP and (2) interactions between NPR-C and NEP increase enzyme activity, we have used purified ovine and human NEP, plus ovine lung plasma membranes to study the relative importance of receptor and enzyme pathways. We have also related the findings to hormone metabolism in vivo. Binding affinities of atrial natriuretic peptide (ANP), hBNP and ovine BNP (oBNP) to oNPR-C were similar (K(d)=8-16 pM). In contrast, unlike ANP and oBNP, hBNP was not significantly degraded by purified oNEP or plasma membranes. Despite similar (and high) affinity of oNPR-C for oBNP and hBNP, the t((1/2)) of hBNP (12.7 min) was more than fourfold that of oBNP (2.6 min). Although we found no evidence for receptor-enzyme interaction, our results show that the delayed metabolism of hBNP reflects resistance to NEP. These findings have important implications for future treatment strategies in human disease. PMID- 11054638 TI - Regulation of rat magnocellular neurosecretory system by D-aspartate: evidence for biological role(s) of a naturally occurring free D-amino acid in mammals. AB - Little evidence is available for the physiological function of D-amino acids in species other than bacteria. Here we demonstrate that naturally occurring freed aspartate (D-Asp) is present in all magnocellular neurons of rat hypothalamus. The levels of this naturally occurring D-amino acid were elevated during lactation and returned to normal thereafter in the magnocellular neurosecretory system, which produces oxytocin, a hormone responsible for milk ejection during lactation. Intraperitoneal injections of D-Asp reproducibly increased oxytocin gene expression and decreased the concentration of circulating oxytocin in vivo. Similar changes were observed in the vasopressin system. These results provide evidence for the role(s) of naturally occurring free D-Asp in mammalian physiology. The findings argue against the conventional concept that only L stereoisomers of amino acids are functional in higher species. PMID- 11054639 TI - Dimethoate inhibits steroidogenesis by disrupting transcription of the steroidogenic acute regulatory (StAR) gene. AB - Dimethoate is a widely used organophosphate insecticide that has been shown to disrupt reproductive function in animals. Although the pathogenesis of Dimethoate induced reproductive toxicity remains to be determined, a reduction in serum testosterone levels is thought to play an important role in the development of Dimethoate-induced infertility. Since Leydig cells play a crucial role in male reproductive function by producing testosterone, the mouse MA-10 Leydig tumor cell line was used to determine if Dimethoate can directly block steroid hormone biosynthesis and to identify the site of steroidogenic inhibition. Dimethoate inhibited steroidogenesis in both a dose- and time-dependent manner without affecting total protein synthesis or protein kinase A activity. While it decreased the activity of the P450 side chain cleavage (P450 scc) enzyme, a reduction in the activity of this enzyme alone could not account for the level of Bu(2)cAMP-inhibited progesterone production. Instead, our results suggest that Dimethoate inhibited steroidogenesis primarily by blocking transcription of the steroidogenic acute regulatory (StAR) gene. This finding is significant since StAR protein mediates the rate-limiting and acutely-regulated step in steroidogenesis, the transfer of cholesterol from the outer to the inner mitochondrial membrane. This study indicates that StAR may be an important target for environmental pollutants which disrupt steroidogenesis and impair reproductive function. PMID- 11054640 TI - Hormonal control of plasmin and tissue-type plasminogen activator activity in rat milk during involution of the mammary gland. AB - We have proposed that growth hormone (GH) and prolactin (PRL) interact to suppress apoptosis in the mammary gland. GH increases insulin-like growth factor I (IGF-I) synthesis whereas PRL suppresses the production of insulin-like growth factor-binding protein-5 (IGFBP-5) in the epithelial cells, which would otherwise inhibit IGF-mediated cell survival. IGFBP-5 was present in milk from involuting glands at high concentrations (approximately 60 microg/ml) and had a high affinity (8.03 x 10(-10) M) for IGF-I, suggesting an inhibitory effect of IGFBP-5 in the mammary gland. IGFBP-5 was present in the micellar fraction of milk and binds specifically to alpha(s2)-casein. Since alpha(s2)-casein also binds plasminogen and tissue-type plasminogen activator (t-PA), resulting in the conversion of plasminogen to plasmin, and since IGFBP-5 binds to plasminogen activator inhibitor-1 (PAI-1), we investigated whether apoptosis and extracellular matrix (ECM) degradation might be coordinately controlled by GH and PRL possibly acting through IGFBP-5. Litters were removed from lactating rats to initiate involution. Plasminogen activation and t-PA activity were both increased dramatically after 48 h and GH and PRL suppressed this response. By contrast, 17beta-oestradiol, progesterone or corticosterone did not influence either process. An antiserum to IGF-I, which blocked systemic IGF-I effects, failed to inhibit the activation of plasminogen or the increase in t-PA, suggesting that paracrine effects of IGF-I may be more important. Teat-sealing, which led to the accumulation of milk without hormonal changes, also led to increases in plasminogen activation and t-PA activity, suggesting that locally produced factors (of which IGFBP-5 is one) are important in controlling ECM remodelling. We propose that GH and PRL inhibit apoptosis and ECM remodelling by a process that involves the control of IGF-I and PAI-1 availability by IGFBP-5, thus allowing these processes to be tightly coordinated. PMID- 11054641 TI - Seasonal-dependent effect of temperature on the response of adenylate cyclase to FSH stimulation in the oviparous lizard, Podarcis sicula. AB - The study of environmental factors affecting vertebrate reproduction has long interested both developmental and evolutionary biologists. Although photoperiod has been considered to be an important environmental parameter for vertebrates such as birds, temperature is probably a primary external factor responsible for reproductive cyclicity in reptiles. In spite of the progress made in the understanding of reptilian reproductive strategies and adaptations, much remains to be learned about the interplay between endocrine physiological factors, such as hormones, and environmental parameters. In this report, we have examined the effects of in vivo administered FSH on oocyte recruitment during the most significant periods of the reproductive cycle of the lizard, Podarcis sicula. The results show that when FSH is administered in proximity to the reproductive period, it stimulates oocyte growth and ovulation; when the hormone is administered at the beginning of the winter stasis it affects ovarian activity without inducing ovulation. Ovarian adenylate cyclase activity is moderately sensitive to in vitro FSH stimulation during the pre- and post-reproductive periods. The sensitivity to hormone stimulation increases significantly during the reproductive period and winter stasis. We have also tested the hypothesis that environmental temperature affects the responsiveness of ovarian adenylate cyclase to FSH stimulation. For such a purpose, we exposed animals to 28 degrees C or 4 degrees C in different periods of the ovarian cycle. The results show that, whenever the temperature applied mimics the thermal regime of the coming season, adenylate cyclase sensitivity to FSH shifts towards levels that anticipate the natural responsiveness. PMID- 11054642 TI - The catechol estrogen, 4-hydroxyestrone, has tissue-specific estrogen actions. AB - Recent data indicate that the catechol estrogen, 2-hydroxyestrone (2-OHE(1)), has no effect on any target tissue including bone, whereas 16 alpha-hydroxyestrone (16 alpha-OHE(1)) exerts tissue-selective estrogen agonist activity. The effect of the catechol estrogen, 4-hydroxyestrone (4-OHE(1)), putatively associated with tumorigenesis, has not been studied in the skeleton. The purpose of this study was to assess the effect of 4-OHE(1) on tibia, uterine and mammary gland histology and blood cholesterol in ovariectomized (OVX'd) growing rats. Ten-week old female Sprague-Dawley rats were injected subcutaneously with 200 microg/kg BW per day with 4-OHE(1), 17 beta-estradiol (E(2)) or vehicle for three weeks. OVX resulted in uterine atrophy, increased body weight, radial bone growth and cancellous bone turnover, and hypercholesterolemia. E(2) prevented these changes with the expected exception that the subcutaneous infusion of this high dose of estrogen did not prevent the hypercholesterolemia. 4-OHE(1) prevented the increase in blood cholesterol and the increase in body weight. 4-OHE(1) appeared to have partial estrogen activity in the uterus; uterine weight and epithelial cell height were significantly greater than the OVX rats but significantly less (twofold) than the E(2) animals. Analysis of variance indicated that 4-OHE(1) slightly decreased the periosteal mineral apposition rate (P<0.05) compared with vehicle-treated rats but had no effect on double-labeled perimeter or bone formation rate. Similarly, 4-OHE(1) was a partial estrogen agonist on cancellous bone turnover. The data suggest that the catechol estrogen, 4-OHE(1), unlike 2 OHE(1), has estrogen activity. Furthermore, the profile of activity differs from that of 16 alpha-OHE(1). Our results suggest that estrogen metabolites may selectively influence estrogen-target tissues and, concomitantly, modulate estrogen-associated disease risk. PMID- 11054643 TI - Regulation of immunoreactive inhibin A and B secretion in cultured human granulosa-luteal cells by gonadotropins, activin A and insulin-like growth factor type-1 receptor. AB - Inhibins are gonadal glycoproteins with endocrine effects on pituitary FSH secretion and para/autocrine effects on ovarian and testicular function. The purpose of this study was to investigate the endocrine and para/autocrine regulation of inhibin A and inhibin B secretion in human ovarian granulosa-luteal cells. The cells were obtained from women undergoing in vitro fertilization, and the primary cultures were treated with FSH, LH, human chorionic gonadotropin (hCG), activin A, 8-bromo cyclic AMP (8-BrcAMP), staurosporine (a protein kinase C inhibitor) and an antagonist of IGF action (type-1 IGF receptor antibody alpha IR3). The secretion of inhibins was measured by ELISA assays capable of reliably distinguishing between inhibin A and B. FSH, LH, hCG and 8-BrcAMP increased inhibin A secretion on average up to 180% (P<0.01), 192% (P<0.05), 210% (P<0.01) and 243% (P<0.01) respectively of the control level, while their stimulatory effect on inhibin B secretion was less pronounced (up to 167%, P<0.01; 139%, P<0.05; 127%, P>0.05; 133%, P>0.05 of the controls respectively). alpha IR3 decreased inhibin A and B secretion down to 70% (P<0.01) and 50% (P<0.01) respectively of the control. Staurosporine decreased inhibin B secretion down to 49% (P<0.01) of the control; its effect on inhibin A secretion was not significant. Activin A increased inhibin B secretion up to fourfold of the control (P<0.05) while its effect on inhibin A secretion was insignificant. We conclude that gonadotropins via the protein kinase A signal transduction pathway are the main positive regulators of inhibin A and B secretion in human granulosa luteal cells. The protein kinase C signal transduction pathway seems to be important especially for inhibin B secretion. Locally produced IGFs are probably important inducers of the production of both forms of inhibin in human ovaries while activins seem to upregulate inhibin B secretion. PMID- 11054644 TI - Dose-response effects of a new growth hormone receptor antagonist (B2036-PEG) on circulating, hepatic and renal expression of the growth hormone/insulin-like growth factor system in adult mice. AB - The effects of growth hormone (GH) in regulating the expression of the hepatic and renal GH and insulin-like growth factor (IGF) system were studied by administering a novel GH receptor antagonist (GHRA) (B2036-PEG) at different doses (0, 1.25, 2.5, 5 and 10 mg/kg/day) to mice for 7 days. No differences were observed in the groups with respect to body weight, food consumption or blood glucose. However, a dose-dependent decrease was observed in circulating IGF-I levels and in hepatic and renal IGF-I levels at the highest doses. In contrast, in the 5 and 10 mg/kg/day GHRA groups, circulating and hepatic transcriptional IGF binding protein-3 (IGFBP-3) levels were not modified, likely resulting in a significantly decreased IGF-I/IGFBP-3 ratio. Hepatic GH receptor (GHR) and GH binding protein (GHBP) mRNA levels increased significantly in all GHRA dosage groups. Endogenous circulatory GH levels increased significantly in the 2.5 and 5 mg/kg/day GHRA groups. Remarkably, increased circulating IGFBP-4 and hepatic IGFBP-4 mRNA levels were observed in all GHRA administration groups. Renal GHR and GHBP mRNA levels were not modified by GHRA administration at the highest doses. Also, renal IGFBP-3 mRNA levels remained unchanged in most GHRA administration groups, whereas IGFBP-1, -4 and -5 mRNA levels were significantly increased in the 5 and 10 mg/kg/day GHRA administration groups. In conclusion, the effects of a specific GHR blockade on circulating, hepatic and renal GH/IGF axis reported here, may prove useful in the future clinical use of GHRAs. PMID- 11054645 TI - Estrogen modulates osteoblast proliferation and function regulated by parathyroid hormone in osteoblastic SaOS-2 cells: role of insulin-like growth factor (IGF)-I and IGF-binding protein-5. AB - Although there is clinical evidence showing that combined therapy with parathyroid hormone (PTH) and estrogen is additively effective in increasing the bone mass of patients with osteoporosis, the mechanism of the interaction between these hormones remains unclear. The present study was performed to determine whether estrogen would affect osteoblast proliferation and function modulated by PTH in human osteoblastic SaOS-2 cells. Human PTH-(1-34) significantly inhibited [(3)H]thymidine (TdR) incorporation, which was attenuated by 24 h pretreatment with 10(-10) to 10(-7) M 17 beta-estradiol (17 beta-E(2)) in a concentration dependent manner. PTH significantly stimulated alkaline phosphatase (ALP) activity, collagen synthesis and type-1 procollagen mRNA expression after pretreatment with 17 beta-E(2 )in these cells. Tamoxifen, an anti-estrogen, antagonized these 17 beta-E(2)-induced effects. Pretreatment with insulin-like growth factor-I (IGF-I) mimicked estrogen action, and coincubation of 3 microg/ml anti-IGF-I antibody antagonized the effects of 17 beta-E(2 )as well as those of IGF-I. In the presence of 17 beta-E(2 )pretreatment, PTH strongly stimulated IGF binding protein (IGFBP)-5 mRNA expression in these cells, and recombinant IGFBP-5 increased type-1 procollagen mRNA expression and ALP activity. In conclusion, estrogen attenuates PTH-induced inhibition of osteoblast proliferation and PTH stimulates osteoblast function in the presence of estrogen pretreatment. IGF-I and/or IGFBP-5 seemed to be involved in the estrogen-induced modulation of PTH action on osteoblast proliferation and function. PMID- 11054646 TI - Metabolic hormones and tissue concentrations of mRNA for IGF-I in lines of sheep that differ in their protein synthesis response to feed intake. AB - The rate of protein synthesis in the skin and muscle of sheep that have been genetically selected for high wool staple strength (SS) is less dependent on the level of dietary intake than that of low SS sheep. This study examined potential hormonal mediators of this difference in responsiveness. Sheep from SS+ and SS- genotypes were fed at 0.4, 1.1 or 1.8 times maintenance. Circulating concentrations of metabolic hormones and tissue concentrations of the mRNA for IGF-I were measured and compared with rates of protein synthesis measured previously. Plasma concentrations of GH, insulin, cortisol, thyroxine and IGF-I responded similarly to dietary intake in both genotypes, but SS+ sheep had higher plasma concentrations of IGF-I at all levels of nutrition (P<0.05). There were no interactions between diet and genotype. The concentration of mRNA for IGF-I was higher in the liver of SS+ sheep (P<0.05), and tended to increase (P=0.06) with nutrient intake, but there were no significant effects of genotype or nutrition in skin, muscle or gut. Concentrations of mRNA for IGF-I were not related to the rate of protein synthesis in any tissue examined. It was concluded that IGF-I did not drive the rate of protein synthesis directly, but it may mediate the responsiveness of protein synthesis rate, or protein degradation rate, to nutrient supply. PMID- 11054647 TI - Prolonged food restriction and mild exercise in Shetland ponies: effects on weight gain, thyroid hormone concentrations and muscle Na(+),K(+)-ATPase. AB - We determined the effects of food supply and low-intensity training on growth, serum thyroid hormone levels and the Na(+),K(+)-pump concentration in equine skeletal muscle. Twenty-two Shetland ponies were subjected to two different feeding regimes for 2(1/2) years (11 ponies per group): food restriction (body condition score kept at 2) or ad libitum fed (body condition score kept at 8). Five ponies in each group underwent low-intensity training. Gluteus medius muscle and serum samples were obtained in April 1998. Subsequently, all ponies were fed ad libitum and the training programme was stopped. Muscle biopsies and serum samples were collected again in November 1998. Food restriction was associated with a 30-50% reduction of body weight gain. While the total thyroxine (T(4)) level was increased, the free T(4) remained at the control level. The serum total tri-iodothyronine (T(3)) and free T(3) were reduced by 30% and 49% respectively. After 6 months of refeeding there were no differences in any of the hormone levels between the ad libitum fed and the food-restricted groups. Food restriction produced a minor, but not significant, decrease in the Na(+),K(+) pump concentration in the gluteus medius muscle of the Shetland ponies. Low intensity training reduced weight gain of the ad libitum fed group by 25%, but had no detectable effect on the concentration of the Na(+), K(+)-pumps. We conclude that prolonged food restriction in Shetland ponies results in a weight gain reduction of 30-50%, and is associated with similar decreases in serum total and free T(3). The reduction in serum T(3) only slightly influenced the Na(+), K(+)-ATPase concentration in skeletal muscle, indicating that muscle tissue of different species may respond differently to changes in circulating thyroid hormones. PMID- 11054648 TI - Gastrointestinal growth factors and pancreatic islet hormones during postoperative IGF-I supplementation in man. AB - Insulin-like growth factor-I (IGF-I) has been demonstrated to exert a nitrogen sparing effect, both experimentally and in patients after abdominal surgery. IGF I is a major mediator for the anabolic effects of growth hormone (GH). Whether elevated circulating IGF-I levels are the sole mediator of the anabolic effects following GH has not been clarified. IGF-I influences glucose metabolism, both through its own specific receptor and by activating the insulin receptor, and has also been proposed to influence pancreatic islet secretion directly. In the present study, the postoperative effects of IGF-I on plasma levels of other gastrointestinal and pancreatic islet hormones and growth factors were measured in patients after abdominal surgery. Fifteen patients who were candidates for large bowel resection were randomly divided into two groups: IGF-I-treated (n=8) and placebo-treated (n=7). The IGF-I group received daily two s.c. injections of human recombinant IGF-I (80 microg/kg body weight) for five days, beginning on the morning of the first postoperative day. The other group received placebo injections. Fasting plasma levels of gastrointestinal growth factors (epidermal growth factor, transforming growth factor-alpha, IGF-II), gastrointestinal hormones (gastrin, enteroglucagon, peptide YY), and islet hormones (insulin, islet amyloid polypeptide (IAPP) and pancreatic glucagon) were determined by RIA preoperatively and after five days of treatment. No significant effects of IGF-I on other growth factors or gastrointestinal hormones were seen. A marked increase in plasma insulin postoperatively compared with the preoperative levels (42+/-3 vs 61+/-5 pM, P<0.05) was seen in the placebo group, whereas the postoperative levels in the IGF-I-treated patients remained unchanged (44+/-3 vs 45+/-4 pM). A similar pattern was observed for IAPP and cortisol concentrations. No differences in glucagon concentrations were seen. In conclusion, these results suggest that IGF-I does not influence production of other gastrointestinal hormones thought to be involved in alimentary growth or pancreatic glucagon. In contrast, IGF-I caused a marked reduction of insulin and IAPP secretion. The inhibition of beta cell secretion could be direct or, alternatively, could involve an improvement in postoperative insulin resistance, perhaps by reducing serum cortisol. PMID- 11054649 TI - Selective modification of the pyruvate dehydrogenase kinase isoform profile in skeletal muscle in hyperthyroidism: implications for the regulatory impact of glucose on fatty acid oxidation. AB - The pyruvate dehydrogenase kinases (PDK1-4) regulate glucose oxidation through inhibitory phosphorylation of the pyruvate dehydrogenase complex (PDC). Immunoblot analysis with antibodies raised against recombinant PDK isoforms demonstrated changes in PDK isoform expression in response to experimental hyperthyroidism (100 microg/100 g body weight; 3 days) that was selective for fast-twitch vs slow-twitch skeletal muscle in that PDK2 expression was increased in the fast-twitch skeletal muscle (the anterior tibialis) (by 1. 6-fold; P<0.05) but not in the slow-twitch muscle (the soleus). PDK4 protein expression was increased by experimental hyperthyroidism in both muscle types, there being a greater response in the anterior tibialis (4.2-fold increase; P<0.05) than in the soleus (3.2-fold increase; P<0.05). The hyperthyroidism-associated up-regulation of PDK4 expression was observed in conjunction with suppression of skeletal muscle PDC activity, but not suppression of glucose uptake/phosphorylation, as measured in vivo in conscious unrestrained rats (using the 2-[(3)H]deoxyglucose technique). We propose that increased PDK isoform expression contributes to the pathology of hyperthyroidism and to PDC inactivation by facilitating the operation of the glucose --> lactate --> glucose (Cori) and glucose --> alanine - > glucose cycles. We also propose that enhanced relative expression of the pyruvate-insensitive PDK isoform (PDK4) in skeletal muscle in hyperthyroidism uncouples glycolytic flux from pyruvate oxidation, sparing pyruvate for non oxidative entry into the tricarboxylic acid (TCA) cycle, and thereby supporting entry of acetyl-CoA (derived from fatty acid oxidation) into the TCA cycle. PMID- 11054650 TI - Renal Na-K-ATPase hyperactivity in diabetic Psammomys obesus is related to glomerular hyperfiltration but is insulin-independent. AB - Psammomys obesus, a desert rodent, develops diabetes when displaced from its natural environment and fed a high energy diet in the laboratory. This study was designed to examine variations in renal function in relation to the diabetic state with emphasis on changes in Na-K-ATPase activity. The following groups of Psammomys were studied: (1) Animals fed a saltbush diet; a low energy/high salt diet (natural). (2) Animals fed a low energy/low salt diet (laboratory). Both 1 and 2 were normoglycemic and normoinsulinemic and thus served as control. (3) Animals fed a high energy diet (group C) who were hyperglycemic and hyperinsulinemic; this group was divided into two subgroups: C1 presented with glomerular hyperfiltration rate and C2 with glomerular hypofiltration rate. (4) Animals fed a high energy diet presenting with hyperglycemia-hypoinsulinemia (group D). (5) Group D+I, similar to group D but treated with external insulin (2 U/24 h). Groups D and C1, whose glomerular filtration rose above normal by 30% and 70% respectively, exhibited metabolic similarity to Type I and Type II diabetes. In these groups, Na-K-ATPase activity in the cortex increased by 80 100% and in the medulla by 180% (P<0.001 vs control). In group C2 with reduced glomerular filtration rate (GFR), Na-K-ATPase activity did not differ from control. In group D+I, with normalized glomerular filtration rate, Na-K-ATPase activity was similar to control. There was a linear and significant correlation between GFR and Na-K-ATPase activity both in the cortex and in the medulla. These experiments present a well defined animal model of diabetes mellitus. Variations in glucose and in insulin did not correlate with Na-K-ATPase activity. These results clearly demonstrated that Na-K-ATPase activity in the diabetic Psammomys was determined by glomerular filtration but was independent of plasma glucose or insulin levels. PMID- 11054651 TI - Selective enhancement of Raman or fluorescence spectra of biomolecules using specifically annealed thick gold films. AB - Annealing of "thick" metal films deposited onto a smooth dielectric substrate leads to high-order self-organization of metal clusters on the film surface. This work presents the first experimental evidence that the "thick" gold film (TGF) may be specifically annealed to serve as a substrate for surface-enhanced fluorescence or surface-enhanced Raman scattering (SERS) spectroscopy of the same molecule. High-resolved SERS spectra of mitoxantrone (mitox) were recorded on the TGF annealed at 340 degrees C whereas no Raman enhancement but an increase of mitox fluorescence signal were detected on the TGF annealed at 240 degrees C. The mitox fluorescence was further enhanced by deposition of monolayers of pentanethiol or poly-L-lysine on the surface of annealed TGF. The maximal fluorescence enhancement factor per mitox molecule of approximately 50 that was obtained on the annealed TGF covered with poly-L-lysine makes the system promising for applications in immunofluorescence assays and in microspectrofluorescence analysis. PMID- 11054652 TI - IR spectroscopic characteristics of cell cycle and cell death probed by synchrotron radiation based Fourier transform IR spectromicroscopy. AB - Synchrotron radiation based Fourier transform IR (SR-FTIR) spectromicroscopy allows the study of individual living cells with a high signal to noise ratio. Here we report the use of the SR-FTIR technique to investigate changes in IR spectral features from individual human lung fibroblast (IMR-90) cells in vitro at different points in their cell cycle. Clear changes are observed in the spectral regions corresponding to proteins, DNA, and RNA as a cell changes from the G(1)-phase to the S-phase and finally into mitosis. These spectral changes include markers for the changing secondary structure of proteins in the cell, as well as variations in DNA/RNA content and packing as the cell cycle progresses. We also observe spectral features that indicate that occasional cells are undergoing various steps in the process of cell death. The dying or dead cell has a shift in the protein amide I and II bands corresponding to changing protein morphologies, and a significant increase in the intensity of an ester carbonyl C===O peak at 1743 cm(-1) is observed. Biopolymers (Biospectroscopy) 57: 329-335, 2000 PMID- 11054653 TI - Structural aspects of thiol-specific spin labeling of human plasma low density lipoprotein. AB - A novel thiol-specific spin labeling procedure for the protein component (apoprotein B, apoB) of low density lipoproteins (LDLs) is presented. A methanethiosulfonate spin label was used to probe the free cysteine residues of apoB with electron paramagnetic resonance (EPR) spectroscopy. The results indicated that the spin labeled sites are predominantly buried in the LDL particle in two distinct environments that differ in their mobility restrictions. The suitability of thiol-specific labeling for the study of the stability and conformation of apoB was demonstrated in experiments with denaturing agents. The results presented in this work offer a new approach for the matching of EPR data with the primary structure of apoB. PMID- 11054654 TI - Quantitation of aliphatic suberin in Quercus suber L. cork by FTIR spectroscopy and solid-state (13)C-NMR spectroscopy. AB - This work determined that the percentage of suberin in cork may be found by solid state (13)C cross polarization/magic angle spinning (CP/MAS) NMR spectroscopy and by FTIR with photoacoustic detection (FTIR-PAS) spectroscopy. A linear relationship is found between the suberin content measured through CP/MAS spectral areas and that measured gravimetrically. Furthermore, application of a partial least squares (PLS1) regression model to the NMR and gravimetric data sets clearly correlates the two sets, enabling suberin quantification with 90% precision. Suberin quantitation by FTIR-PAS spectroscopy is also achieved by a PLS1 regression model, giving 90% accurate estimates of the percentage of suberin in cork. Therefore, (13)C-CP/MAS NMR and FTIR-PAS proved to be useful and accurate noninvasive techniques to quantify suberin in cork, thus avoiding the traditional time consuming and destructive chemical methods. PMID- 11054655 TI - Raman and IR spectroscopic investigation of zinc(II)-carnosine complexes. AB - The zinc(II)-L-carnosine system was investigated at different pH and metal/ligand ratios by Raman and IR spectroscopy. The Raman and IR spectra present some marker bands useful to identify the sites involved in metal chelation at a specific pH value. In particular, the neutral imidazole group gives rise to some Raman bands, such as the nu C(4)===C(5) band, that change in wave number, depending on whether the imidazole ring takes the tautomeric form I or II. Even if tautomer I is predominant in the free ligand, metal coordination can upset tautomeric preference and N(tau)- and N(pi)-ligated complexes can be identified. Although weak compared to those of aromatic residues, these Raman marker bands may be useful in analyzing metal-histidine interaction in peptides and proteins. On the basis of the vibrational results, conclusions can be drawn on the species existing in the system. Depending on the available nitrogen atoms, various complexes can be formed and the prevalent form of the species depends mainly on the pH. At basic pH carnosine gives rise to two different neutral complexes: a water-insoluble polymeric species, [ZnH(-1)L](0)(n), and a dimer, [Zn(2)H( 2)L(2)](0). The first is predominant and involves the tautomeric I form of the imidazole ring in metal chelation; the second contains tautomer II and increases its percentage by going from a 2 to 0.25 metal/ligand ratio. Conversely, the dimeric species dominates at pH 7, whereas two charged species, [ZnHL](2+) and [ZnL](+), are formed under slightly acidic conditions. In the [ZnHL](2+) complex the imidazole ring takes part in the Zn(II) coordination in the tautomeric I form, whereas in [ZnL](+) the ring is protonated and not bound to the Zn(II) ion. In addition, the curve fitting analysis of the 1700-1530 cm(-1) Raman region was helpful in indicating the predominant species at each pH. PMID- 11054656 TI - Chemiluminescent method for determination of tetracycline, chlortetracycline, minocycline, doxycycline, and demeclocycline. AB - A chemiluminescent method is described for the determination of tetracycline, chlortetracycline, minocycline, doxycycline, and demeclocycline. The method is based on the photon counting technique and the spline functions approximation. The simple formula S = b + Ax(2) is proposed for the observed dependency of the integrated number of countings S on the concentration x of a given antibiotic. The correlation between S and the half-life of a drug in the human body is proved. PMID- 11054657 TI - New nonbonded interactions calculation strategy for rectangular systems. I. Preliminary molecular dynamics study of solvated Na(+) ion. AB - A new molecular nonbonded interactions treatment strategy is proposed in the context of rectangular periodic boundary conditions simulations. Several molecular dynamics simulations are performed on a sodium ion in aqueous solution. Box sizes are modified from a cubic to a rectangular shape. The results are compared with those found using a classical spherical cutoff. This new method yields ion-oxygen radial distribution functions in good agreement with experimental results, thus showing its reliability. Severe perturbations in the structural orientation of water molecules in the first shell with the increase of the box length are observed under the classical cutoff method. However, these distorting effects are reduced with the present nonbonded interactions treatment. PMID- 11054658 TI - DNA adducts as markers of exposure to carcinogens and risk of cancer. PMID- 11054659 TI - Salivary carcinoma in HER-2/neu transgenic male mice: an angiogenic switch is not required for tumor onset and progression. AB - Morphologic examinations of salivary gland neoplasias arising in male BALB/c (H 2d) mice carrying the activated HER-2/neu (BALB-NeuT) indicate that expression of the oncogene product in the ductal-acinar structures results in a very human-like acinic cell adenocarcinoma with a smoldering course and infrequent metastatization. Typical and then atypical hyperplasia of ducts and acini preceded the rise of salivary tumors that originated from the confluence of multiple ductal hyperplastic foci, while hyperplastic acini behaved as an abortive preneoplastic lesion. The vascular network in normal, hyperplastic and neoplastic salivary tissue was analysed to see whether activation of the angiogenic process is essential in salivary gland carcinogenesis. Immunostaining with anti-endothelial cells (anti-CD31), anti-beta3 integrin and anti-laminin antibodies revealed that microvessel density was significantly higher in normal and hyperplastic than in neoplastic tissue, in which no signs of new vessel sprouting were found. Assessment of angiogenic factor expression indicates a low presence of VEGF in normal, hyperplastic and neoplastic epithelium, while bFGF was preferentially produced but not exported by neoplastic cells and remained in a cell-associated form. Our data suggest that normal salivary gland vascularization is able to support tumor onset and development with no need for an angiogenic switch. PMID- 11054660 TI - IGFBP-3 prolongs the p53 response and enhances apoptosis following UV irradiation. AB - Neoplastic transformation is characterised by an imbalance in favour of cell growth over programmed cell death (apoptosis). The tumour-suppressor gene p53, responsible for maintaining cell-cycle control, is mutated in the majority of human cancers. Loss of function of the target genes of p53 are therefore important in tumourigenesis. One such target gene is the insulin-like growth factor binding protein-3 (IGFBP-3), an extracellular protein responsible for the carriage of IGF-I but which can act independently of IGF-I, inhibiting cell growth and enhancing apoptosis. Using the KYSE 190 oesophageal carcinoma cell line, we have demonstrated that IGFBP-3 alone has no effect on cell growth or cell survival. However, it significantly enhanced apoptosis, with a 67% increase in the pre-G1 peak on flow cytometry following UV irradiation. The increase in p53 was enhanced and prolonged when cells are stressed in the presence of IGFBP 3. These data suggest an autocrine/paracrine feedback loop exists between IGFBP-3 and p53, which may provide the social control necessary to maintain normal tissue homeostasis. PMID- 11054661 TI - Acetaldehyde production by non-pathogenic Neisseria in human oral microflora: implications for carcinogenesis in upper aerodigestive tract. AB - Many epidemiological studies have identified chronic alcohol consumption as a significant risk factor for cancer of the upper aerodigestive tract (UAT) in human. Although acetaldehyde, the first metabolite from ethanol by alcohol dehydrogenase (ADH), is regarded as a carcinogen, how systemic production of acetaldehyde particularly affects the UAT remains unclear. In our study, we searched for the regional source of acetaldehyde in UAT, especially the involvement of bacteria in the human normal oral microflora. Here we demonstrate that, among the bacterial species identified from the human oral cavity, genus Neisseria had extremely high ADH activity and produced significant amounts of acetaldehyde when cultured with medium containing ethanol in vitro. The ability to produce acetaldehyde was more than 100-fold higher than that produced by any other genera we studied. Furthermore, alcohol ingestion influences the bacterial composition of the oral microflora, resulting in an increased proportion of Neisseria. Although Neisseria present in normal oral microflora is generally non pathogenic, these findings suggest that this microbe can be a regional source of carcinogenic acetaldehyde and thus potentially play an important role in alcohol related carcinogenesis in human UAT. PMID- 11054662 TI - Negative regulation of alkylation-induced sister-chromatid exchange by poly(ADP ribose) polymerase-1 activity. AB - One of the earliest responses to DNA damage in eukaryotic cells is activation of poly(ADP-ribose) polymerase-1 (PARP-1), a DNA strand break-dependent nuclear enzyme which covalently modifies proteins with poly(ADP-ribose). Here, we show that conditional over-expression of PARP-1 in stably transfected hamster cells, which causes cellular over-accumulation of poly(ADP-ribose) by several-fold, strongly suppresses alkylation-induced sister-chromatid exchange (SCE), while cytotoxicity of alkylation treatment is slightly enhanced. Viewed together with the known potentiation of SCE by abrogation of PARP-1 activity, our results provide evidence that PARP-1 activity is an important regulator of alkylation induced SCE formation, imposing a control that is strictly negative and commensurate with the level of enzyme activity. PMID- 11054663 TI - Down-regulation of the Diphthamide biosynthesis protein 2-like gene during retinoid-induced differentiation and apoptosis: implications against its tumor suppressor activity. AB - Retinoids, synthetic and natural analogs of retinoic acid (RA) have profound effects on the proliferation and differentiation of many cell types; this accounts for their beneficial effects in the treatment of certain neoplasias. We have employed mRNA differential display to characterize genes associated with differentiation and apoptosis induced by all-trans RA in human lung cancer cells. We have identified a cDNA corresponding to the sequence of the known gene diphthamide biosynthesis protein 2-like (DPH2L). Although the function of this gene remains unknown, as it was first isolated from the critical region of deletion on chromosome 17p13.3 in human ovarian carcinoma, it has been regarded as a candidate tumor-suppressor gene. In this report, we provide evidence that DPH2L is down-regulated during differentiation or apoptosis in several cancer cell lines after treatment with all-trans RA or N-(4-hydroxyphenyl)retinamide and during cell-cycle arrest. Moreover, stable expression of DPH2L-specific anti sense construct leads to inhibition of cell proliferation. Our results suggest that DPH2L in not a conventional tumor-suppressor gene. Instead, it may be a growth regulator and its down-regulation might be permissive for the transition from cell growth to differentiation or apoptosis. DPH2L might be a useful tool in the prognosis of neoplastic diseases. PMID- 11054664 TI - Up-regulation of the protein tyrosine phosphatase SHP-1 in human breast cancer and correlation with GRB2 expression. AB - The protein tyrosine phosphatase SHP-1 is predominantly expressed in hemopoietic cell lineages, where its function is relatively well defined. However, its expression profile also extends to certain epithelial cell types. Furthermore, the negative regulatory role of this enzyme in hemopoietic cell signaling may not apply to other systems, where positive effects on particular tyrosine kinase signaling pathways have been described. Expression of SHP-1 was therefore investigated in human breast cancer cell lines and primary breast cancers. Differential expression of SHP-1 mRNA was observed among the 19 breast cancer cell lines examined, and in an analysis of 72 primary breast cancers, SHP-1 mRNA expression was increased 2- to 12-fold relative to normal breast epithelial cells in 58% of the samples. Interestingly, a subset of the cancers also over-expressed GRB2 mRNA by 2- to 7-fold, and a significant (p < 0.01) positive correlation was observed between SHP-1 and GRB2 mRNA expression. Since these proteins can bind to each other and regulate MEK/MAP kinase activation, their co-ordinate up regulation may amplify tyrosine kinase signaling in breast cancer cells. PMID- 11054665 TI - Tiam1 mutations in human renal-cell carcinomas. AB - Tiam1 activates the Rho-like GTPase Rac1, and studies indicate that Tiam1-Rac1 signaling affects invasion in different ways depending on the cell type studied. However, no investigations on Tiam1 in human tumors have been reported. Here, we show that for 4 of 5 human renal-cell carcinoma (RCC) cell lines the expression levels of Tiam1 tended to be inversely correlated with in vitro invasiveness, whereas no obvious correlation could be found between the expression levels of Rac1 and invasion. Subsequent mutation analysis of these cell lines revealed no mutations in Rac1 but up to 5 different point mutations in the Tiam1 gene. Of these, 1 mutation (A441G) was located in the NH2-terminal pleckstrin homology domain, which is essential for membrane localization and functional activity of Tiam1. By analysis of an additional 30 primary human RCCs, mutation A441G was found in 4 of 35 tumors and tumor cell lines (11.5%) but not in the respective normal kidney tissues. By enzymatic digestion, mutation A441G proved to be heterozygous, suggesting a dominant active function. This was supported by showing that stable over-expression of mutated A441G-Tiam1 induced transformation of NIH3T3 cells, as determined in a colony formation assay, whereas empty vector and wild-type Tiam1 failed to do so. In conclusion, a distinct Tiam1 mutation (A441G) was identified in several human RCCs. This mutation induced transformation of NIH3T3 cells and, hence, might play a major role in the progression of human RCCs. Further analyses on Tiam1 mutations in human tumors might give new clues to their role in tumor progression. PMID- 11054666 TI - Over-expression of p27kip1 induces growth arrest and apoptosis mediated by changes of pRb expression in lung cancer cell lines. AB - p27kip1 is a cyclin-dependent kinase inhibitor which controls the G1 phase of the cell cycle in conjunction with pRb. p27 has been associated with cell-cycle arrest and apoptosis. In this study, we transferred the full-length human p27 cDNA using a replication-deficient recombinant adenoviral vector (Ax-p27) into lung cancer cell lines and evaluated the potential of this strategy for anti cancer gene therapy. After infection with Ax-p27, the growth of H322, A549 and SQ 5 cells, which express pRb, was almost completely suppressed, though no such effect was found in H69 and Lu-135 cells, which do not express pRb. In addition, cell death from day 4 after infection with Ax-p27 was observed only in H322, A549 and SQ-5 cells but not in H69 and Lu-135 cells. The cell cycle of H322 cells treated with Ax-p27 became arrested at the G1 phase from day 1 to day 3 despite continued over-expression of p27. When we examined the changes in expression level of pRb and E2F-1, which play important roles in cell-cycle progression from G1 to S phase, down-regulation of pRb expression was detected in H322 cells 3 days after infection with Ax-p27. These data suggest that (i) the growth inhibitory effect and induction of apoptosis by over-expression of p27 require expression of pRb and (ii) adenovirus-mediated p27 gene transfer may have promise as a novel strategy in cancer gene therapy. PMID- 11054667 TI - Overexpression of the csk gene suppresses tumor metastasis in vivo. AB - The non-receptor tyrosine kinase c-Src has been implicated in the development of numerous human cancers. c-Src is activated in colon cancers, particularly in highly metastatic cells, and its overexpression strongly correlates with tumor progression. C-terminal Src kinase (Csk) has been demonstrated to negatively regulate Src family tyrosine kinases through tyrosine phosphorylation at the C terminal regulatory site (Tyr-527). We report herein that down-regulation of Src kinase activity by adenovirus-mediated csk gene transfer abrogated the highly metastatic phenotype of colon cancer cells. Overexpression of Csk decreased Src tyrosine kinase activity in NL-17 cells, the highly metastatic clone of mouse colon adenocarcinoma 26. Importantly, Csk overexpression in NL-17 cells resulted in significant suppression of in vivo metastasis, without affecting its tumorgenicity. Csk overexpression decreased the invasiveness of NL-17 cells through Matrigel, in vitro reconstituted basement membrane. Gelatin zymography confirmed the decreased protein levels of MMP-2 (gelatinase A) in the supernatants of Csk-overexpressed NL- 17 cells. These results provide a therapeutic basis for interfering with metastasis of colon cancer by csk gene mediated down-regulation of Src kinase activity. PMID- 11054668 TI - Over-expression of nucleophosmin/B23 decreases the susceptibility of human leukemia HL-60 cells to retinoic acid-induced differentiation and apoptosis. AB - Stable clones of HL-60 cells in which nucleophosmin/B23 was over-expressed were established. Less percentages (4-20%) of nucleophosmin/B23 over-expressed (pCR3 B23) cells exhibited the morphological characteristic of apoptosis as compared with control vector-transfected (pCR3) cells (6-53%) during the 10 microM RA treatment for 1-4 days. In flow cytometry analysis, a block in the G1 phase was noted in all the pCR3-B23 and pCR3 cells after 2 days of 10 microM RA treatment and continued to be observed at all times measured up to 6 days. Smaller peaks of apoptotic cells with less than G1 DNA content were observed in pCR3-B23 as compared with pCR3 cells after 4-6 days of 10 microM RA treatment. As measured by expressions of differentiation markers and the functional assessment of the ability to reduce nitroblue-tetrazolium, our results further showed that over expression of nucleophosmin/B23 decreased the response of the cells to RA-induced differentiation. Less cleavage of PARP and in vitro caspase-3 activity were observed in PCR3-B23 cells as compared with pCR3 cells treated with 10 microM RA for 3-4 days. IRF-1 was induced after 6 hr of 10 microM RA treatment in the pCR3 B23 and pCR3 cells. Significantly more nucleophosmin/B23 was co immunoprecipitated with IRF-1 from pCR3-B23 cells than from pCR3 cells during RA treatment (10 microM; 24 hr, 96 hr). The IRF-1 transcriptional activity was found to be attenuated in pCR3-B23 cells as compared with pCR3 cells during the treatment of cells with RA. Nucleophosmin/B23, through interacting with IRF-1, plays an important role in the control of the susceptibility of cells to RA induced differentiation and apoptosis. PMID- 11054669 TI - Induced neuroblastoma cell differentiation, associated with transient HES-1 activity and reduced HASH-1 expression, is inhibited by Notch1. AB - Neuroblastoma is a childhood tumor that originates from the sympathetic nervous system. The tumor cells have embryonic features, presumably as a consequence of an impaired capacity to respond to signals and transcriptional control mechanisms operating during normal differentiation. Two basic helix-loop-helix transcription factors, human achaete-scute homologue-1 (HASH-1) and hairy/enhancer of split homologue-1 (HES-1), are crucial for proper development of some neuronal cells. Here, their potential roles during sympathetic differentiation of human neuroblastoma cells have been investigated. In all tested protocols for induction of differentiation of SH-SY5Y and SK-N-BE(2) neuroblastoma cells, HASH-1 expression was rapidly decreased with a concomitant, often transient, increase in HES-1 expression. In gel mobility shift assays, using extracts from neuroblastoma cells, HES-1 bound to an oligonucleotide corresponding to a sequence in the HASH 1 promoter including the so-called N-box, suggesting that the transiently increased HES-1 activity in differentiating neuroblastoma cells is involved in down-regulation of HASH-1. Constitutive expression of the intracellular domain of Notch1, which activates the HES-1 promoter in SH-SY5Y cells, inhibited spontaneous and induced morphological differentiation of these neuroblastoma cells. Our data show that functional sympathetic neuronal differentiation of neuroblastoma cells is associated with transient activation of HES-1 and down regulation of HASH-1 expression. PMID- 11054670 TI - Detection of telomerase activity and correlation with mitotic and apoptotic indices, Ki-67 and expression of cyclins D1 and A in cutaneous melanoma. AB - Telomerase plays a key role in carcinogenesis. It is activated in most immortal cell lines and human cancers, including cutaneous melanoma (CM). Increased cell proliferation and deregulation of the cell cycle occur in human cancers. Links between telomerase activity (TA), cell proliferation, cell death and expression of cell-cycle regulators have not been extensively elucidated in CM. In this study, we investigated TA, mitotic index (MI), apoptotic index (AI), Ki-67 and nuclear positivity of cyclins D1 and A (Ki-67+ N/1,000, cyclin D1+N/1,000, cyclin A+N/1,000) in 42 primary cutaneous melanomas (PCMs). TA was detected in all cases and directly correlated with MI, Ki-67+N/1,000, cyclin D1+N/1,000 and cyclin A+N/1,000 (p < 0.001); it was not correlated with AI. When subdividing PCMs into radial and vertical growth phase melanomas (RGPMs, VGPMs), a correlation was maintained only with MI (p < 0.005) and cyclin D1 +N/1,000 (p < 0.005). Although MI and Ki-67+N/1,000 were highly correlated with cyclin D1+N/1,000 and cyclin A+N/1,000 (p < 0.001) when considering all cases together, a high correlation was found in the RGPM and VGPM groups between cyclin A+N/1,000 and Ki-67+N/1,000 only (p < 0.001), thus suggesting that cyclin A is more closely correlated with cell proliferation than cyclin D1. Our results further support the association between TA, tumor cell proliferation and cyclin D1 and A expression in PCM, though it is possible that links between TA and proliferation, on the one hand, and TA and cyclin D1 expression, on the other, might occur following various pathways. PMID- 11054671 TI - Expression of collagenase-3 (matrix metalloproteinase-13) in transitional-cell carcinoma of the urinary bladder. AB - Expression of collagenase-3 [matrix metalloproteinase-13 (MMP-13)] has been previously demonstrated in squamous-cell carcinomas of both the head and neck and the vulva, cutaneous basal-cell carcinomas, chondrosarcomas and melanomas. Using in situ hybridization, MMP-13 mRNA expression was detected in 13 of 23 (52%) urinary bladder transitional-cell carcinomas (TCCs). Expression was restricted to cells in the invading edges of tumors. No expression of MMP-13 mRNA could be detected in normal urothelium. As detected by immunohistochemistry, MMP-13 protein showed an expression pattern similar to that of MMP-13 mRNA. Expression of MMP-13 mRNA and protein was also detected in 2 bladder carcinoma cell lines (RT4 and T24). In these cell lines, TNF-alpha potently induced MMP-13 mRNA expression. Retinoids and a selective p38 inhibitor, SB203580, potently inhibited MMP-13 mRNA expression. Our results demonstrate MMP-13 expression in human urinary bladder carcinoma cells in vivo and in vitro and suggest that MMP-13 may serve as a marker for transformation and invasion in urinary bladder TCCs. PMID- 11054672 TI - Genetics of chemical carcinogenesis: analysis of bidirectional selective breeding inducing maximal resistance or maximal susceptibility to 2-stage skin tumorigenesis in the mouse. AB - We report on bidirectional selective breeding, initiated from a genetically defined foundation population and carried out to selection limit, for producing lines of mice endowed with maximal resistance (Car-R) or maximal susceptibility (Car-S) to 2-stage skin tumorigenesis. The initial population resulted from a balanced intercrossing of 8 inbred strains of mice. The tumors, induced by a single application of DMBA (initiation) and twice weekly applications of TPA (promotion), were benign papillomas; their number at the end of the promotion period was the phenotype chosen for assortative mating. Afterward, the majority of them regressed while others progressed to malignant carcinomas. The Car-R line was selected through a strong challenge, while the Car-S line selection was based on responses to decreasing concentrations of DMBA and TPA. The selection limit was reached after 14 or 15 generations showing progressive interline divergence, which strongly suggests the interaction of several quantitative trait loci (QTL). The phenotypic difference was extremely large: the tumor response was 73 times higher in Car-S than in Car-R mice, though the applied concentrations of DMBA and TPA were 100 and 40 times lower, respectively. The mean heritability realized during the selective breeding was 0.20 in Car-R and 0.49 in Car-S. Our results are compatible with a minimal QTL estimate of 8 in the Car-R line and of 9 or 10 in the Car-S line. The Car-S line is also much more susceptible to carcinoma induction. An association of coat color with tumorigenesis was observed in interline F2 segregants. The Car-R and Car-S lines, obtained through a long lasting breeding program, are a unique model for identifying the QTL involved in chemical tumorigenesis and will be provided to interested investigators. PMID- 11054673 TI - Transporter (TAP)- and proteasome-independent presentation of a melanoma associated tyrosinase epitope. AB - The melanosomal protein tyrosinase is considered as a target of specific immunotherapy against melanoma. Two tyrosinase-derived peptides are presented in association with HLA-A2.1 [Wolfel et al., Eur. J. Immunol., 24, 759-764 (1994)]. Peptide 1-9 (MLLAVLYCL) is generated from the putative signal sequence. The internal peptide 369-377 is posttranslationally converted at residue 371, and its presentation is dependent on functional TAP transporters and proteasomes [Mosse et al., J. exp. Med.187, 37-48 (1998)]. Herein, we report on the processing and transport requirements for the signal sequence-derived peptide 1-9 that were studied in parallel to those for peptide 369-377. After infection of TAP deficient (T2) and TAP-positive (T1) cells with a Modified Vaccinia Ankara construct carrying the human tyrosinase gene (MVA-hTyr), we found that recognition by CTL against peptide 1-9 did not require TAP function as opposed to recognition by CTL against peptide 369-377. When target cells with intact processing and transport functions were infected with MVA-hTyr, lysis by CTL against peptide 1-9 was not impaired by lactacystin, a specific inhibitor for the proteasome, whereas lysis by CTL against peptide 369-377 was completely abrogated. Taken together, peptide 1-9 derived from the signal sequence of tyrosinase is presented in a TAP-independent fashion and does not require proteasomes for processing. Cellular immune responses against this hydrophobic peptide can be monitored with lymphokine spot assays as documented in the case of a patient with metastatic melanoma, in whom we observed a preferential T-cell response against tyrosinase peptide 1-9 subsequent to chemoimmunotherapy. Independence of cytosolic processing and transport pathways and potentially enhanced expression levels make signal sequence-derived peptides and their carrier proteins important candidates for specific immunotherapy. PMID- 11054674 TI - SH2D1A and SLAM protein expression in human lymphocytes and derived cell lines. AB - The gene defect responsible for the X-linked lymphoproliferative disease (XLP) is associated with an impaired control of Epstein-Barr virus (EBV) infection. The gene has been recently identified and the encoded protein (designated SH2D1A, DSHP or SAP) was characterized. It is a 128 amino acid (aa) protein, containing a single Src homology 2 (SH2) domain. It interacts with signaling lymphocytic activation molecule (SLAM) expressed on the surface of activated T and B cells. We show that activated T, but not activated B, cells express the SH2D1A protein. NK cells express the protein as well. Tumor lines originating from B, T or NK cells exhibited similar SH2D1A protein expression as the corresponding normal cells, with some notable exceptions. EBV-carrying, tumor phenotype representative (type I), but not EBV-carrying lymphoblastoid cell line (LCL)-like (type III) or EBV-negative Burkitt lymphoma (BL) lines expressed SH2D1A. The phenotypic switch from type I to type III in the EBV-carrying BL line Mutu was associated with a down-regulation of SH2D1A and up-regulation of SLAM. In contrast to normal ex vivo and long-term activated NK cells, 2 of 3 NK leukemia lines expressed SLAM. All 3 lines expressed SH2D1A, like their normal counterparts. PMID- 11054675 TI - Insulin-like growth factor binding protein-3 (IGFBP-3) potentiates paclitaxel induced apoptosis in human breast cancer cells. AB - Variability in response to chemotherapy is poorly understood. Paclitaxel-induced apoptosis was assessed in human Hs578T breast cancer cells, using the MTT assay, cell counting, morphological features and flow cytometry. Pre-dosing cells with non-glycosylated insulin-like growth factor binding protein-3 (ngIGFBP-3) had no effect on the cells per se but accentuated paclitaxel-induced apoptosis. The apoptotic pathway was further examined by measuring caspase-3 activity in cell lysates at time points over 48 hr after dosing with paclitaxel. Activity increased significantly, and Western immunoblots for caspase-3 in conditioned media showed that the inactive precursor decreased after incubation with paclitaxel. Endogenous production of IGFBP-3 by the cells after incubation with paclitaxel was evaluated using Western ligand blotting, specific IGFBP-3 immunoblotting and radioimmunoassay. Paclitaxel increased endogenous IGFBP-3, which was further increased if the cells had been pre-dosed with ngIGFBP-3. These findings suggest that IGFBP-3 may be an important modulator of paclitaxel-induced apoptosis. PMID- 11054676 TI - Oncolytic potential of E1B 55 kDa-deleted YKL-1 recombinant adenovirus: correlation with p53 functional status. AB - YKL-1, E1B 55 kDa-deleted recombinant adenovirus vector, capable of harboring a transgene casette of up to 4.9 kb, was newly constructed by reintroducing E1A and E1B 19 kDa into E1/E3-deleted adenoviral vector with a homologous recombination in E. coli. Virus replication and cytotoxicity were dramatically attenuated in all 3 different types of normal human cells. In contrast, YKL-1 efficiently replicated and induced cytotoxicity in most cancer cells, especially Hep3B and C33A cells with an inactivating p53 mutation. However, both H460 and HepG2 exhibited intermediate sensitivity to YKL-1, which was between that of Hep3B or C33A and normal human cells. The YKL-1 and DNA damaging agent, camptothecin effectively induced p53 in H460 and HepG2 as well as in normal cells. Furthermore, YKL-1 effectively prohibited both Hep3B and C33A tumor growth in nu/nu mice in a dose-dependent manner. H/E staining and TUNEL assay indicated a largely distributed necrotic area and apoptosis on its periphery. This study, therefore, indicates that YKL-1, possesses promising potential as an oncolytic adenoviral vector, which acts partially in a p53-dependent manner. PMID- 11054677 TI - Regulation of gamma-glutamyltransferase in cisplatin-resistant and -sensitive colon carcinoma cells after acute cisplatin and oxidative stress exposures. AB - Glutathione plays an important role in drug resistance of tumor cells and in their ability to resist oxidative stress. Improved salvage of glutathione can be obtained through increased activity of gamma-glutamyltransferase (GGT), which is of importance in the maintenance of cellular glutathione homeostasis. We investigated the regulation of GGT in 2 cisplatin-resistant and 1 cisplatin sensitive colon carcinoma cell lines. Enzyme activity was induced in all 3 cell lines after acute exposure to cisplatin. The elevation was significantly higher in sensitive cells (3.3-fold) than in resistant (1.6- to 1.7-fold) cells. Exposure of cells to oxidative stress generated by menadione also resulted in enzyme induction but only in cisplatin-sensitive cells. Addition of anti-oxidants had different effects on the 2 inductions: N-acetylcysteine blocked the induction of both cisplatin and menadione, whereas catalase and glutathione-ester blocked only the menadione induction. Glutathione depletion alone was not sufficient to induce GGT in these cells. The data show that GGT is regulated by multiple mechanisms during anti-tumor drug treatment and oxidative stress and that reactive oxygen species were involved in the menadione, but not cisplatin, induction of the enzyme. PMID- 11054678 TI - O6-methylguanine formation, repair protein depletion and clinical outcome with a 4 hr schedule of temozolomide in the treatment of advanced melanoma: results of a phase II study. AB - O6-Methylguanine-DNA methyltransferase (MGMT) is a major determinant of resistance to temozolomide. Its levels are depleted in lymphocytes after drug administration, but there is partial recovery by 24 hr, the usual time of subsequent dosing. Administering subsequent doses of temozolomide at the MGMT nadir could enhance its effectiveness, by increasing the amount of O6 methylguanine (O6-meG) in DNA. We evaluated the efficacy of such a schedule of temozolomide and determined the kinetics of MGMT depletion and O6-meG formation in DNA following treatment. Thirty patients with advanced malignant melanoma were treated with temozolomide 1,000 mg/m2 equally split into 5 doses over a 16 hr period every 28 days. O6-meG formation was determined in peripheral blood mononuclear cell (PBMC) DNA and, in a subset of patients, in tumor tissue during the first treatment cycle. MGMT levels fell rapidly with dosing, reaching a nadir in PBMCs of 18.0 +/- 2.26% of initial levels. O6-meG levels increased during the treatment period, peaking at 11.1 +/- 1.25 micromol/mol dG in PBMCs and at 4.25 +/- 0.79 micromol/mol dG in tumor biopsies. The main toxicities were grade IV thrombocytopenia in 12 patients (42.8%) and grade IV neutropenia in 11 patients (39.2%), associated with fever in 8 cases. There were 7 responses (1 complete), for an overall response rate of 23.3%; median overall survival was 6.1 months. The compressed schedule has activity against melanoma, with greater MGMT depletion and O6-meG formation than previously reported for O6-alkylating agent regimens. Myelosuppression precludes its wider application, but MGMT in PBMCs predicted the dose intensity of temozolomide that patients could sustain, suggesting a means by which individuals suitable for this approach might be identified. PMID- 11054679 TI - Cisplatin combined with prostaglandin E1 chemotherapy in rat peritoneal carcinomatosis. AB - Cisplatin intraperitoneal (i.p.) chemotherapy is frequently performed for patients with peritoneal carcinomatosis. However, cisplatin penetrates only the surface of the peritoneal tumor and has serious side effects on renal cells. Thus, cisplatin i.p. chemotherapy had been limited to use for these patients. Prostaglandin E1 (PGE1) has been used for reducing the toxic effects of anticancer drugs because of its cytoprotective effects and has been reported to enhance tumoricidal activity of anticancer drugs. In our study, the effects of PGE1 on the rat peritoneal carcinomatosis model treated with cisplatin i.p. chemotherapy were evaluated. Cisplatin (5 mg/kg) was given in an i.p. administration to 70 tumor-free rats. PGE1 was administered to 35 rats through the tail vein at an infusion rate of 0.1 microg/kg/min (1 ml/hr), and the remaining 35 rats were injected with physiological saline. Forty rats were given an i.p. injection of 1 x 10(7) AH100B cells. Ten days after injection, cisplatin (5 mg/kg) was administered with PGE1 to 20 and the remaining 20 were injected with physiological saline. The accumulation of platinum in the tissues and apoptotic renal cells were analyzed. The maximum concentrations of platinum in the kidneys of PGE1 untreated rats (tumor-free: 10.11 microg/g; tumor-bearing: 11.45 microg/g) did not differ from those of platinum in the kidneys of PGE1 treated rats (tumor-free: 10.28 microg/g; tumor-bearing: 13.28 microg/g). The number of apoptotic renal cells was significantly reduced by PGE1 administration in both tumor-free and tumor-bearing rats. Moreover, PGE1 increased the maximum platinum concentration in tumor masses (5.31 microg/g) of the treated group compared with that in tumor mass of the control group (2.72 microg/g, p = 0.009). These results indicate that PGE1 may increase the anticancer effect of cisplatin by increasing tumor platinum concentration and may reduce the chance of cisplatin induced renal failure. Intraperitoneal cisplatin chemotherapy combined with PGE1 treatment may have a therapeutic benefit for patients with peritoneal carcinomatosis. PMID- 11054680 TI - Circadian organization of thymidylate synthase activity in normal tissues: a possible basis for 5-fluorouracil chronotherapeutic advantage. AB - Fluoropyrimidines induce cytotoxicity, in part, by inhibiting the proliferation coordinated enzyme thymidylate synthase (TS), which is essential for DNA synthesis. Tumor TS levels are clinically predictive of post-surgical tumor recurrence and of response to fluoropyrimidine chemotherapy. Fluoropyrimidine drug toxicity and efficacy each vary reproducibly in humans and animals, depending upon their circadian timing. In vivo, normal tissues and some tumor tissues exhibit circadian coordination of cellular proliferation. We therefore asked whether TS activity is coordinated rhythmically throughout the day in the normal proliferative tissues most damaged by fluoropyrimidine drugs. To assess tissue and time of day TS activity differences, we harvested normal tissues from female mice living on a 12:12 hr light:dark schedule at each of 6 different equispaced times throughout a 24 hr cycle and measured TS catalytic activity. We observed up to 10-fold differences in vivo in TS activity among different normal tissue types, roughly paralleling their proliferative state and relative fluoropyrimidine sensitivity. In normal tissues most damaged by fluoropyrimidines (bone marrow, small intestinal mucosa and oral mucosa/tongue), TS activity varies up to 2-fold throughout each day. In bone marrow, the circadian pattern of TS activity parallels the circadian rhythm in proliferation in this tissue. This circadian organization of TS, one of the primary fluoropyrimidine targets in normal tissues, probably contributes in vivo to the time of day differences in the toxic-therapeutic ratio of circadian-timed fluoropyrimidine drug therapy. PMID- 11054681 TI - Neonatal level of thyroid-stimulating hormone and acute childhood leukemia. AB - One of the more consistent findings in leukemia research is the association between birth weight and childhood leukemia. Because thyroid hormones are critically involved in growth and differentiation, we speculated that hormone levels could be of significance to the development of leukemia in early life. Specifically, we hypothesized that high levels of thyroid-stimulating hormone (TSH) would be associated with a low risk of leukemia. Accordingly, high TSH (low free thyroid hormone) early after birth most likely reflects low function of the thyroid accompanied by low rate of cell turnover and so lower risk of faulty cell divisions leading to cancer. In a matched case-control study nested from all singleton children born in Denmark between 1986 and 1998, we compared levels of TSH (as measured in a neonatal screening program for congenital hypothyroidism) in 188 cases of acute lymphoblastic leukemia (ALL) and 28 of acute myeloid leukemia (AML) with levels in 1,450 and 216 matched controls, respectively. Data were analyzed using conditional logistic regression and odds ratios (OR) were adjusted for birth weight. As hypothesized, we found a decreased risk of ALL and AML associated with high TSH (OR(ALL) = 0.7 [0.5-1.0]; OR(AML) = 0.3 [0.1-1.0]). However, both conditions were also associated with low levels of TSH (OR(ALL) = 0.4 [0.2-0.7]; OR(AML) = 0.3 [0.1-1.4]). In conclusion, extreme TSH levels a few days after birth appears to be associated with a decreased risk of acute childhood leukemia. PMID- 11054682 TI - The spectrum of HIV-1 related cancers in South Africa. AB - Despite the high prevalence of infection by the Human Immunodeficiency Virus (HIV) in South Africa, information on its association with cancer is sparse. Our study was carried out to examine the relationship between HIV and a number of cancer types or sites that are common in South Africa. A total of 4,883 subjects, presenting with a cancer or cardiovascular disease at the 3 tertiary referral hospitals in Johannesburg, were interviewed and had blood tested for HIV. Odds ratios associated with HIV infection were calculated by using unconditional logistic regression models for 16 major cancer types where data was available for 50 or more patients. In the comparison group, the prevalence of HIV infection was 8.3% in males and 9.1% in females. Significant excess risks associated with HIV infection were found for Kaposi's sarcoma (OR=21.9, 95% CI=12.5-38.6), non Hodgkin lymphoma (OR=5.0, 95%CI=2.7-9.5), vulval cancer (OR=4.8, 95%CI= 1.9-12.2) and cervical cancer (OR= 1.6, 95%CI= 1.1-2.3) but not for any of the other major cancer types examined, including Hodgkin disease, multiple myeloma and lung cancer. In Johannesburg, South Africa, HIV infection was associated with significantly increased risks of Kaposi's sarcoma, non-Hodgkin lymphoma and cancers of the cervix and the vulva. The relative risks for Kaposi's sarcoma and non-Hodgkin lymphoma associated with HIV infection were substantially lower than those found in the West. PMID- 11054683 TI - Mutagen sensitivity in patients with familial and non-familial urothelial cell carcinoma. AB - Due to variation in individual susceptibility, only a fraction of all individuals exposed to environmental carcinogens will develop cancer. Our aim was to assess whether mutagen sensitivity plays a role in developing urothelial cell carcinoma (UCC) and whether this sensitivity is different in familial and non-familial cases. Intrinsic susceptibility was quantified by a mutagen sensitivity assay (mean number of chromatid breaks per cell after damage induction with bleomycin in the late S-G2 phase of the cell cycle). Patients were classified as sporadic (n = 25), familial (2 patients in 1 nuclear family, n = 23) or hereditary (2 patients <60 years or 3 patients in 1 nuclear family, n = 13) and compared with control subjects without a history of cancer. Information on demographic factors, smoking history and family history of UCC was collected by postal questionnaires. Differences in mutagen sensitivity were assessed by ANOVA and logistic regression analysis. Overall, UCC patients showed a higher mutagen sensitivity score compared with control subjects [mean number of chromatid breaks per cell 0.91, 95% confidence interval (CI) 0.84-0.97, and 0.74, 95% CI 0.69-0.79, respectively; p = 0.001). Sporadic and familial patients exhibited the highest susceptibility (0.94, 95% CI 0.82-1.06, and 0.93, 95% CI 0.83-1.03, respectively). Hereditary patients (0.79, 95% CI 0.72-0.86) showed a susceptibility similar to controls. Mutagen sensitivity increases the risk of non-hereditary UCC. The relatively low mutagen sensitivity score among hereditary patients points to a different carcinogenic pathway. PMID- 11054684 TI - Risk of malignancy among patients with rheumatic conditions. AB - Previous studies have described an increased risk of malignancy in subjects diagnosed with rheumatic conditions, most notably rheumatoid arthritis (RA). Our aim was to quantify and compare risks for site-specific malignancy among hospitalized patients with RA, osteoarthritis (OA) and other rheumatic conditions in a nationwide, population-based cohort. Subjects were identified from Scottish hospital in-patient records from 1981 to 1996 and followed up by computer linkage of the Scottish Cancer Registry and the national registry of deaths. Expected cancer incidence was calculated from national cancer rates and related to the observed incidence by the standardized incidence ratio (SIR). Among RA patients, there was an increased risk for hematopoietic [males SIR= 2.13, 95% confidence interval (CI) 1.7-2.7; females SIR = 1.76, 95% CI 1.5-2.1], lung (males SIR = 1.32, 95% CI 1.2-1.5; females SIR = 1.44, 95% CI 1.3-1.6) and prostate (SIR = 1.26, 95% CI 1.0-1.6) cancers. Reduced risk were seen for colorectal cancer (males SIR = 0.87, 95% CI 0.7-1.1; females SIR = 0.71, 95% CI 0.6-0.9) and, among females, stomach cancer (SIR = 0.70, 95% CI 0.5-1.0). The excess risk for hematopoietic cancer and the reduced risk for colorectal and stomach cancers were sustained over 10 years of follow-up. An overall decreased risk of cancer was observed for patients with OA; the greatest reductions were observed for colorectal (males SIR = 0.88, 95% CI 0.8-1.0; females SIR = 0.84, 95% CI 0.8 0.9), stomach (males SIR = 0.79, 95% CI 0.7-0.9; females SIR = 0.66, 95% CI 0.6 0.8) and lung (males SIR = 0.72, 95% CI 0.7-0.8; females SIR = 0.84, 95% CI 0.8 0.9) malignancies, with decreased risks generally still evident at 10 years of follow-up. Our results support several previous findings regarding the incidence of hematopoietic and colorectal malignancies in RA patients. In addition, we have shown a large decrease in stomach cancer among patients with OA and females with RA that warrants further investigation since it may provide clues to possible prevention strategies. To further our knowledge about the underlying mechanisms of altered risk in cancer patients with rheumatic conditions, population studies requiring primary data collection are required. PMID- 11054685 TI - The Von Hippel-Lindau (VHL) tumor-suppressor gene is not mutated in sporadic human colon adenocarcinomas. PMID- 11054686 TI - Synaptic proteins in rat taste bud cells: appearance in the Golgi apparatus and relationship to alpha-gustducin and the Lewis(b) and A antigens. AB - Taste receptor cells are continuously replaced during the life of the animal, but many of their sensory axons respond primarily to stimuli belonging to a single taste quality. This suggests that a newly arising taste cell must form a synapse with an appropriate sensory axon, requiring cell recognition that is likely to be mediated by surface markers. As an approach to studying this process, we attempted to locate synapses by immunolabeling taste buds of rats for proteins involved in neurotransmitter release. In taste bud cells of vallate papillae and nasoincisor ducts, double-labeling experiments showed that syntaxin-1, SNAP-25, synaptobrevin, and synaptophysin colocalized with the Golgi marker beta COP in elongated cytoplasmic compartments that extended from the perinuclear region into apical and basal processes of the cells. Labeled cells were spindle-shaped, identifying them as light cells. Syntaxin-1 appeared only in taste cells, but SNAP-25, synaptobrevin, and synaptophysin were also seen in nerve fibers. The synaptic vesicle glycoprotein SV2 appeared only in nerve fibers. Taste cells of fungiform papillae did not show immunoreactivity for presynaptic proteins or Golgi markers, but axonal labeling was similar to that in other regions. Taste cells with alpha-gustducin could express either presynaptic proteins or the carbohydrate blood group antigen Lewis(b), but not both. Therefore, Lewis(b) and presynaptic proteins are not expressed during the same period in the life of a taste bud cell. Most taste cells expressing syntaxin-1 (82%) also expressed the A blood group antigen, whether or not they expressed alpha-gustducin. PMID- 11054687 TI - Estrogen-regulated progestin receptors are found in the midbrain raphe but not hippocampus of estrogen receptor alpha (ER alpha) gene-disrupted mice. AB - Estrogen and progesterone may modulate serotonergic function through intracellular receptors, alpha (ER alpha) and/or beta (ER beta), and the progestin receptor (PR). Studies in macaque and rat suggest species differences in steroid action. Presently, we examined the mouse. To identify whether ER alpha is involved in estrogen induction of PR in midbrain raphe, we studied the ER alpha gene-disrupted (alpha ERKO) mouse. The hippocampus was examined as another estrogen/progestin-sensitive brain area reported to express ER alpha, ER beta, and PR. Female and male homozygous alpha ERKO and wildtype mice were gonadectomized and given estradiol benzoate or vehicle. Dual-label immunocytochemistry was performed for PR or ER alpha and the serotonin synthesizing enzyme, tryptophan hydroxylase (TPH). Cells exhibiting PR immunoreactivity (PR-ir) or ER alpha-ir were observed in dorsal and median raphe and hippocampus in both sexes. No ER alpha-ir cells were observed in alpha ERKO brains. In raphe, PR-ir or ER alpha-ir often colocalized with TPH-ir. Thus, estrogen and progesterone may directly modulate gene expression in select serotonergic neurons via ER alpha and PR in female and male mice. Estrogen significantly increased the number of PR-ir cells, and the percentage of PR-ir cells colocalizing TPH-ir in both raphe nuclei, regardless of sex and genotype. Although less among alpha ERKO mice, the significant estrogen induction of PRs implicates the involvement of another ER, perhaps ER beta. In hippocampus, distinct estrogen-induced PR-ir cells were observed only in wildtype animals, demonstrating an ER alpha-mediated event in this forebrain region. Collectively, these findings suggest that estrogen can regulate the expression of one gene (the PR) via multiple mechanisms, based upon brain region. PMID- 11054688 TI - Differential expression of glutamate receptor subtypes in human brainstem sites involved in perinatal hypoxia-ischemia. AB - This study delineates the development of N-methyl-D-aspartate (NMDA) and non-NMDA receptor binding in the human brainstem, particularly as it relates to issues of the trophic effects of glutamate, the glutamate-mediated ventilatory response to hypoxia, and regional excitotoxic vulnerability to perinatal hypoxia-ischemia. We used tissue autoradiography to map the development of binding to NMDA, alpha amino-3-hydroxy-5-methyl-4-isoxazole-proprionate (AMPA), and kainate receptors in brainstem sites involved in the glutamate ventilatory response to hypoxia, as well as recognized sites vulnerable to perinatal hypoxia-ischemia. NMDA receptor/channel binding was virtually undetectable in all regions of the human fetal brainstem at midgestation, an unexpected finding given the trophic role for NMDA receptors in early central nervous system maturation in experimental animals. In contrast, non-NMDA (AMPA and kainate) receptor binding was markedly elevated in multiple nuclei at midgestation. Although NMDA binding increased between midgestation and early infancy to moderately high adult levels, AMPA binding dramatically fell over the same time period to low adult levels. High levels of kainate binding did not change significantly between midgestation and infancy, except for an elevation in the infant compared with fetal inferior olive; after infancy, kainate binding decreased to negligible adult levels. Our data further suggest a differential development of components of the NMDA receptor/channel complex. This baseline information is critical in considering glutaminergic mechanisms in human brainstem development, physiology, and pathology. PMID- 11054689 TI - Abnormal retinotopic organization of the dorsal lateral geniculate nucleus of the tyrosinase-negative albino cat. AB - Visual defects associated with hypopigmentation have been studied extensively in Siamese and albino cats. Previous research on tyrosinase-negative albino cats has shown that (1) approximately 95% of all nasal and temporal retinal fibers cross at the optic chiasm, and (2) ocular dominance columns normally found in cortex are replaced with hemiretinal domains. In this study, we compared the retinotopic organization of the dorsal lateral geniculate nucleus (LGNd) and visual cortex in albino cats. Extracellular recordings were conducted in the LGNd, area 17, and area 18 of six albino cats. Receptive fields (RFs) were plotted for all sites. We find that, as in albino visual cortex, the albino LGNd contains (1) normal cells with RFs in the visual hemifield contralateral to the recording site (RFc), (2) abnormal cells with RFs in the ipsilateral hemifield (RFi), (3) abnormal cells with dual, mirror-symmetric RFs, one in each hemifield (RFd), and (4) abnormal cells with broad RFs that span the vertical meridian (RFb). Our data indicate that lamina A and lamina A1 consist predominantly of normal RFc and abnormal RFi cells, respectively. The C laminae contain a mixture of RFc, RFi, RFd, and RFb cells. The interlaminar zones contained RFd cells, RFb cells, or both. Thus, the albino LGNd is arranged into hemiretinal and not ocular dominance laminae. Finally, the percentage of normal cells is significantly larger in area 17 (84%) and area 18 (70%) than in the LGNd (46%), suggesting a suppression of abnormal activity in albino cat cortex, which could underlie the existing competence of visual function in albinos. PMID- 11054690 TI - Ultrastructural localization of the serotonin transporter in superficial and deep layers of the rat prelimbic prefrontal cortex and its spatial relationship to dopamine terminals. AB - Dopamine levels within the prefrontal cortex (PFC) can be manipulated by selective inhibitors of the serotonin transporter (SERT). However, the cellular mechanisms underlying these effects are not clear. The present study sought to examine the distribution of immunogold-silver labeling for SERT (SERT-ir) in the rat prelimbic PFC and to describe its ultrastructural spatial relationship to dopamine axons labeled by immunoperoxidase staining for tyrosine hydroxylase (TH ir). SERT was localized to axonal profiles that ranged in size from fine caliber fibers containing dense SERT-ir, primarily along the membrane, and rarely forming synapses to large, spherical varicosities exhibiting less dense staining, mainly within the cytoplasm, and more commonly forming synapses. Synaptic contacts of SERT profiles were typically asymmetric, axospinous, and more frequent in superficial (38%) than deep (19%) layers. For TH-ir profiles, only 24% were within the same 13.8 microm(2) microenvironment as SERT-ir profiles. Furthermore, TH-ir and SERT-ir profiles were rarely directly apposed to each other or convergent onto common dendritic structures. Instead, these two profiles were typically separated by an average distance of 1.30 microm in the coronal plane, a value that did not vary with the size of SERT-ir axons, the amount of SERT labeling, or the cortical layer examined. These results are consistent with two populations of SERT profiles within the rat prelimbic PFC that may arise from different raphe nuclei or that represent varicose and intervaricose portions of the same axons. Moreover, the functional interactions between cortical serotonin and dopamine systems that may contribute to the therapeutic efficacy of antidepressant drugs are likely to occur over distances greater than 1 microm. PMID- 11054691 TI - Regional and cellular distribution of protein kinase C in rat cerebellar Purkinje cells. AB - Protein kinase C (PCK) is a family of isoforms that are implicated in subcellular signal transduction. The authors investigated the distribution of several PKC isoforms (PKC-alpha, PKC-beta, PKC-gamma, PKC-delta, and PKC-epsilon) within major cerebellar cell types as well as cerebellar projection target neurons, including Purkinje neurons, cerebellar nuclear neurons, and secondary vestibular neurons. PKC-alpha, PKC-beta, PKC-gamma, PKC-delta, and PKC-epsilon are found within the cerebellum. Of these isoforms, PKC-gamma and PKC-delta are highly expressed in Purkinje cells. PKC-gamma is expressed in all Purkinje cells, whereas the expression of PKC-delta is restricted to sagittal bands of Purkinje cells in the posterior cerebellar cortex. In the lower folia of the uvula and nodulus, Purkinje cell expression of PKC-delta is uniformly high, and the sagittal banding for PKC-delta expression is absent. Within the cerebellar nuclei, PKC-delta-immunolabeled axons terminate within the medial aspect of the caudal half of the ipsilateral interpositus nucleus. PKC delta-immunolabeled axons also terminated within the caudal medial and descending vestibular nuclei (MVN and DVN, respectively), the parasolitary nucleus (Psol), and the nucleus prepositus hypoglossi (NPH). PKC-gamma-immunolabeled axons terminated in all of the cerebellar nuclei as well as in the lateral and superior vestibular nuclei and the MVN, DVN, Psol, and NPH. The projection patterns of PKC-immunolabeled Purkinje cells were confirmed by lesion-depletion studies in which unilateral uvula-nodular lesions caused depletion of PKC-immunolabeled terminals ipsilateral to the lesion in the vestibular complex. These data identify circuitry that is unique to cerebellar-vestibular interactions. PMID- 11054692 TI - Relationship between vestibular primary afferents and vestibulospinal neurons in lampreys. AB - The distribution of vestibular primary afferents as well as their relationship with vestibulospinal and other brainstem neurons were studied in lampreys using anatomical tracers. Afferents from the anterior (aVIIIn) and the posterior (pVIIIn) branches of the vestibular nerve were located mainly in the ventral nucleus of the octavolateral area. The relationship between afferents and vestibulospinal neurons was studied by applying one fluorescent tracer to the whole vestibular nerve or one of its branches and applying another tracer to the spinal cord. Some afferents showed large, bulb-like enlargements (bulbs) and about 20 of these were found in the anterior and the intermediate octavomotor nucleus, whereas about 40 were found in the posterior octavomotor nucleus. Some of the bulbs made apparent contact with vestibulospinal neurons in the intermediate octavomotor nucleus and originated mostly from the aVIIIn, whereas bulbs in the posterior octavomotor nucleus originated from the pVIIIn. Applications of biocytin to hemisegments of rostral spinal cord labeled vestibulospinal neurons located in the ipsilateral intermediate octavomotor nucleus and the contralateral posterior octavomotor nucleus. In addition, vestibular primary afferents with bulbs in apparent contact with vestibulospinal neurons were transneuronally labeled by biocytin. They were observed in the ipsilateral aVIIIn and the contralateral pVIIIn and could be followed in the labyrinths, where they innervated the vertical and horizontal arms of the semicircular canal crests. Taken together, these results indicate that vestibular primary afferents from the aVIIIn innervate predominantly vestibulospinal neurons of the intermediate octavomotor nucleus, whereas afferents from the pVIIIn innervate vestibulospinal neurons in the posterior octavomotor nucleus. This anatomical organization suggests that afferents carrying bulbs convey dynamic information to vestibulospinal neurons, which, in turn, project to the spinal cord networks. PMID- 11054693 TI - Distribution of NADPH-diaphorase reactivity and effects of nitric oxide on feeding and locomotory circuitry in the pteropod mollusc, Clione limacina. AB - The action of nitric oxide (NO) and the distribution of putative nitric oxide synthase-containing cells in the pelagic pteropod mollusc Clione limacina were studied using nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry and conventional microelectrode techniques in the isolated central nervous system and in semi-intact preparations. The majority of NADPH-d-reactive neuronal somata were restricted to the cerebral ganglia. The labeled cells were small in diameter (20-30 microm) and were located in the medial areas of the ganglia. A pair of symmetrical neurons was found in the peripheral "olfactory organ." NADPH-d-reactive non-neuronal cells were detected in the periphery and were mainly associated with secretorylike cells and organs of the renopericardial system. The NO donor, diethylamine NO complex sodium salt (10-100 microM), activated neurons from both feeding and locomotory circuits. The cGMP analog, 8 Br-cGMP, mimicked the effects of NO on neurons. We suggest that NO is an endogenous neuromodulator involved in the control of some aspects of feeding and locomotor behavior of Clione. PMID- 11054694 TI - Is there tonic activity in the endogenous opioid systems? A c-Fos study in the rat central nervous system after intravenous injection of naloxone or naloxone methiodide. AB - This study examined the possibility that a tonic activity in the endogenous opioid systems (EO systems) exists in animals under normal conditions. In a first set of experiments, concurrent changes in behavioral responses and in the numbers of c-Fos-like immunoreactive (Fos-LI) neurons in 58 structures of the brain and lumbosacral spinal cord were analyzed in rats after systemic administration of the opioid antagonist naloxone (NAL; 2 mg/kg). Possible roles of the EO systems were inferred from changes in the numbers of Fos-LI neurons between normal rats that received either NAL or the same volume of saline. Free-floating sections were processed immunohistochemically for c-Fos protein using standard avidin biotin complex methods. After NAL, the numbers of Fos-LI neurons were significantly increased in the area postrema; in the caudal, intermediate, and rostral parts of the nucleus tractus solitarii; in the rostral ventrolateral medulla; in the Kolliker-Fuse nucleus; in the supramammillary nucleus; and in the central nucleus of the amygdala. In a second set of experiments examining changes in c-Fos expression in the latter structures, similar increases were found after NAL but not after an equimolar dose of NAL-methiodide, a preferential, peripherally acting opioid receptor antagonist. Therefore, Fos-LI was likely triggered after blockade of central opioid receptors, but not peripheral opioid receptors, releasing neurons from EO system-mediated inhibition. The results of this study suggest the existence of a tonic activity of the EO systems exerted on a restricted number of brain regions in normal rats. This tonic activity of the EO systems may control part of the neural networks involved in cardiorespiratory functions and in emotional and learning processes. PMID- 11054695 TI - Auditory thalamocortical projections in the cat: laminar and areal patterns of input. AB - Thalamocortical projections were studied in adult cats using biotinylated dextran amines, wheat germ agglutinin conjugated to horseradish peroxidase, and autoradiography with tritiated leucine and/or proline. The input from 7 architectonically defined nuclei to 14 auditory cortical fields was characterized qualitatively and quantitatively. The principal results were that 1) every thalamic nucleus projected to more than 1 field (range, 4-14 fields; mean, 7 fields); 2) only the projection from the ventral division to some primary fields (primary auditory cortex and posterior auditory cortex) had a periodic, clustered distribution, whereas the input from other divisions to nonprimary areas was continuous; 3) layers III-V received >85% of the total axonal profiles; 4) in most experiments, five or more layers were labeled; 5) the projections to nonprimary auditory areas had many laterally oriented axons; 6) the heaviest input to layer I in all experiments was usually in its upper half, suggesting a sublaminar arrangement; 7) the largest axonal trunks (up to 6 microm in diameter) arose from the medial division and ended in layer Ia, where they ran laterally for long distances; 8) there were three projection patterns: type 1 had its peak in layers III-IV with little input to layer I, and it arose from the ventral division and the dorsal superficial, dorsal, and suprageniculate nuclei of the dorsal division; type 2 had heavy labeling in layer I and less in layers III-IV, arising from the dorsal division nuclei primarily, especially the caudal dorsal and deep dorsal nuclei; and type 3 was a trimodal concentration in layers I, III IV, and VI that originated chiefly in the medial division and had the lowest density of labeling; and 9) the quantitative profiles with the three methods were very similar. The results suggest that the subdivisions of the auditory thalamus have consistent patterns of laminar distribution to different cortical areas, that an average of five or more layers receive significant input in a specific area, that a given thalamic nucleus can influence areas as far as 20 mm apart, that the first information to arrive at the cortex may reach layer I by virtue of the giant axons, and that several laminar patterns of auditory thalamocortical projection exist. The view that the auditory thalamus (and perhaps other thalamic nuclei) serves mainly a relay function underestimates its many modes for influencing the cortex on a laminar basis. PMID- 11054696 TI - Appreciating the difference between design-based and model-based sampling strategies in quantitative morphology of the nervous system. AB - Quantitative morphology of the nervous system has undergone great developments over recent years, and several new technical procedures have been devised and applied successfully to neuromorphological research. However, a lively debate has arisen on some issues, and a great deal of confusion appears to exist that is definitely responsible for the slow spread of the new techniques among scientists. One such element of confusion is related to uncertainty about the meaning, implications, and advantages of the design-based sampling strategy that characterize the new techniques. In this article, to help remove this uncertainty, morphoquantitative methods are described and contrasted on the basis of the inferential paradigm of the sampling strategy: design-based vs model based. Moreover, some recommendations are made to help scientists judge the appropriateness of a method used for a given study in relation to its specific goals. Finally, the use of the term stereology to label, more or less expressly, only some methods is critically discussed. PMID- 11054697 TI - Movement of mitochondria in the axons and dendrites of cultured hippocampal neurons. AB - Mitochondria generate ATP and are involved in the regulation of cytoplasmic calcium levels. It is thought that local demand for mitochondria differs between axons and dendrites. Moreover, it has been suggested that the distribution of both energy need and calcium flux in dendrites changes with patterns of synaptic activation, whereas the distribution of these demands in axons is stable. The present study sought to determine whether there are differences in mitochondrial movements between axons and dendrites that may relate to differences in local mitochondrial demand. We labeled the mitochondria in cultured hippocampal neurons with a fluorescent dye and used time-lapse microscopy to examine their movements. In both axons and dendrites, approximately one-third of the mitochondria were in motion at any one time. In both domains, approximately 70% of the mitochondria moved in the anterograde direction, whereas the remainder moved in the retrograde direction. The velocity of the movements in each direction in each domain ranged from 0.1 microm/sec to approximately 2 microm/sec, and the means and distributions of the velocities were similar. Only one difference in the behavior of mitochondria between axons and dendrites emerged from this analysis. Mitochondria in axons were more likely to move with a consistently rapid velocity than were those in dendrites. As a result, mitochondria in axons tended to travel farther than mitochondria in dendrites. These results suggest that the transport of mitochondria in axons and dendrites is similar despite any differences in mitochondrial demand between the two domains. PMID- 11054698 TI - Role of microtubules and actin filaments in the movement of mitochondria in the axons and dendrites of cultured hippocampal neurons. AB - The mitochondria in the axons and dendrites of neurons are highly motile, but the mechanism of these movements is not well understood. It has been thought that the transport of membrane-bounded organelles in axons, and perhaps also in dendrites, depends on molecular motors of the kinesin and dynein families. However, recent evidence has suggested that some organelle transport, including that of mitochondria, may proceed along actin filaments as well. The present study sought to determine the extent to which mitochondrial movements in neurons depend on microtubule-based and actin-based transport systems. The mitochondria in cultured hippocampal neurons were labeled with a fluorescent dye and the cells were treated with either nocodazole, a drug that disrupts the microtubule network or cytochalasin D or latrunculin B, drugs which disrupt the actin network. The movement of the mitochondria in the axons and dendrites of neurons after each of these drug treatments was then examined with time-lapse microscopy. Treatment with nocodazole, which depolymerizes microtubules, stopped most mitochondrial movements in both axons and dendrites. Treatment with cytochalasin D, which aggregates actin filaments, also inhibited most movements of mitochondria, but latrunculin B, which depolymerizes actin filaments, had virtually no effect. Together, these data suggest that most of the mitochondrial movements in both axons and dendrites are microtubule-based, but in each domain there may also be some movement along actin filaments. PMID- 11054699 TI - Binocular competition does not regulate retinogeniculate arbor size in fetal monkey. AB - Binocular interactions play a prominent role in shaping the axonal arbors of geniculocortical fibers and the arbors of Y cells in the retinogeniculate pathway of the fetal cat. Fiber interactions between the two eyes have also been suggested to regulate the formation of retinal projections to the dorsal lateral geniculate nucleus (dlgn) of the fetal monkey, but whether this reflects structural refinements of retinal arbors has not been established. To address this issue, we quantified the morphologic properties of individual fibers in two macaque monkeys at embryonic day (E) 110 and E121 that had an eye removed at E69 and E61, respectively. Fibers were labeled by DiI crystals into the fixed optic tract and were visualized by confocal microscopy. Three measurements were made: the number of branch points within the axon terminal arbor, the total arborization length, and the incidence of axonal side branches on the preterminal axon within the confines of the geniculate. There were no significant differences with respect to these parameters between the prenatal enucleates and normal monkeys of comparable age. This was the case for retinal fibers innervating the magnocellular and the parvocellular segments of the dlgn. The arbors stemming from the remaining eye were widely distributed in the dlgn, with some terminating in territories normally innervated by the other (enucleated) eye. These results lend support to the hypothesis that the expanded projection from the remaining eye to the lateral geniculate nucleus of the prenatally enucleated monkey is due to the maintenance of a contingent of retinal fibers normally eliminated by ganglion cell death. PMID- 11054700 TI - Response of abducens internuclear neurons to axotomy in the adult cat. AB - The highly specific projection of abducens internuclear neurons on the medial rectus motoneurons of the oculomotor nucleus constitutes an optimal model for investigating the effects of axotomy in the central nervous system. We have analyzed the morphological changes induced by this lesion on both the cell bodies and the transected axons of abducens internuclear neurons in the adult cat. Axotomy was performed by the transection of the medial longitudinal fascicle. Cell counts of Nissl-stained material and calretinin-immunostained abducens internuclear neurons revealed no cell death by 3 months postaxotomy. Ultrastructural examination of these cells at 6, 14, 24, and 90 days postaxotomy showed normal cytological features. However, the surface membrane of axotomized neurons appeared contacted by very few synaptic boutons compared to controls. This change was quantified by measuring the percentage of synaptic coverage of the cell bodies and the linear density of boutons. Both parameters decreased significantly after axotomy, with the lowest values at 90 days postlesion ( approximately 70% reduction). We also explored axonal regrowth and the possibility of reinnervation of a new target by means of anterograde labeling with biocytin. At all time intervals analyzed, labeled axons were observed to be interrupted at the caudal limit of the lesion; in no case did they cross the scar tissue to reach the distal part of the tract. Nonetheless, a conspicuous axonal sprouting was present at the caudal aspect of the lesion site. Structures suggestive of axonal growth were found, such as large terminal clubs, from which short filopodium-like branches frequently emerged. Similar findings were obtained after parvalbumin and calretinin immunostaining. At the electron microscopy level, biocytin-labeled boutons originating from the sprouts appeared surrounded by either extracellular space, which was extremely dilated at the lesion site, or by glial processes. The great majority of labeled boutons examined were, thus, devoid of neuronal contact, indicating absence of reinnervation of a new target. Altogether, these data indicate that abducens internuclear neurons survive axotomy in the adult cat and show some form of axonal regrowth, even in the absence of target connection. PMID- 11054701 TI - Discharge characteristics of axotomized abducens internuclear neurons in the adult cat. AB - The aim of the present work was to characterize the axotomy-induced changes in the discharge properties of central nervous system neurons recorded in the alert behaving animal. The abducens internuclear neurons of the adult cat were the chosen model. The axons of these neurons course through the contralateral medial longitudinal fascicle and contact the medial rectus motoneurons of the oculomotor nucleus. Axotomy was carried out by the unilateral transection of this fascicle (right side) and produced immediate oculomotor deficits, mainly the incapacity of the right eye to adduct across the midline. Extracellular single-unit recording of abducens neurons was carried out simultaneously with eye movements. The main alteration observed in the firing of these axotomized neurons was the overall decrease in firing rate. During eye fixations, the tonic signal was reduced, and, on occasion, a progressive decay in firing rate was observed. On-directed saccades were not accompanied by the high-frequency spike burst typical of controls; instead, there was a moderate increase in firing. Similarly, during the vestibular nystagmus, neurons hardly modulated during both the slow and the fast phases. Linear regression analysis between firing rate and eye movement parameters showed a significant reduction in eye position and velocity sensitivities with respect to controls, during both spontaneous and vestibularly induced eye movements. These firing alterations were observed during the 3 month period of study after lesion, with no sign of recovery. Conversely, abducens motoneurons showed no significant alteration in their firing pattern. Therefore, axotomy produced long-lasting changes in the discharge characteristics of abducens internuclear neurons that presumably reflected the loss of afferent oculomotor signals. These alterations might be due to the absence of trophic influences derived from the target. PMID- 11054702 TI - Connectivity of the tectal zones coding for upward and downward oblique eye movements in goldfish. AB - Deep layers of the goldfish tectum code movements in a topographically ordered motor map. This work studies the relationship between tectal sites (coding eye movements with different vertical directions) and the distributions of boutons (left by their projections), within rostral mesencephalic structures and rhombencephalic reticular formations. These regions have been involved in the generation of the vertical and horizontal components of eye movement, respectively, as suggested by the Cartesian hypothesis of de-codification of tectal signal. With this aim, discrete injections of biotinylated dextran amine (BDA) and Fluoro-Ruby (FR) were made into functionally identified tectal sites, coding oblique eye movements with similar amplitude of the horizontal component but opposite upward and downward vertical directions, and the distribution of synaptic endings was determined. The main findings of the present work were as follows: 1) within the tectal descending tract, axons were organized according to the location of injected sites within the tectum; 2) BDA and FR boutons were distributed in separate clusters within the medial longitudinal fasciculus and oculomotor nuclei, as well as in the nearby mesencephalic reticular formation; and 3) the regions containing both types of bouton overlapped moderately within the mesencephalic reticular formation at the isthmus level. Overlapping was more extended at the different levels of the rhombencephalic reticular formation, although a shift in the distribution of both types of bouton was always observed. These results suggest that, within the vertical generator, the endings were separated to contact the different neuronal population that codes the upward and downward components of movements. In contrast, in the horizontal generator, tectal endings more likely converge on the same neuronal population to code the horizontal component of movements, irrespective of whether the oblique movements were directed upward or downward. PMID- 11054703 TI - Neurons expressing NADPH-diaphorase in the developing human spinal cord. AB - The present study used nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase histochemistry to identify populations of neurons containing nitric oxide synthase and to describe their putative migration during development of the human spinal cord. As early as week 6 (W6) of gestation, diaphorase expression was observed in sympathetic preganglionic neurons (SPNs) and interneurons of the ventral horn. As development proceeded, the SPNs translocated dorsally to form the intermediolateral nucleus, and the interneurons remained scattered throughout the ventral horn. In addition to the dorsal translocation of SPNs, a unique dorsomedially directed migratory pathway was observed. At later stages of development, other groups of SPNs were identified laterally in the lateral funiculus and medially in the intercalated and central autonomic regions. In addition, two "U-shaped" groups of diaphorase-labeled cells were identified around the ventral ventricular zone at W7. Cells of these groups appeared to translocate dorsally over the next weeks and presumably give rise to interneurons within the deep dorsal horn and surrounding the central canal. Furthermore, during W7-14 of gestation, the deep dorsal horn contained a number of diaphorase positive cells, whereas the superficial dorsal horn was relatively free of staining. These data demonstrate that nitric oxide is present very early in human spinal cord development and that two unique cell migrations initially observed in rodents have now been identified in humans. Furthermore, nitric oxide may be expressed in some populations of neurons as they migrate to their final positions, suggesting that this molecule may play a role in neuronal development. PMID- 11054704 TI - Distribution of mRNAs encoding the arylhydrocarbon receptor, arylhydrocarbon receptor nuclear translocator, and arylhydrocarbon receptor nuclear translocator 2 in the rat brain and brainstem. AB - Dioxin exposure alters a variety of neural functions, most likely through activation of the arylhydrocarbon receptor (AhR) pathway. Many of the adverse effects, including disruption of circadian changes in hormone release and depressed appetite, seem to be mediated by hypothalamic and/or brainstem neurons. However, it is unclear whether these effects are direct or indirect, because there have been no comprehensive studies mapping the expression of components of the AhR pathway in the brain. Therefore, we used a sensitive in situ hybridization histochemical (ISHH) method to map the neural expression of AhR mRNA, as well as those of the mRNAs encoding the AhR dimerization partners, arylhydrocarbon receptor nuclear translocator (ARNT) and ARNT2. We found that AhR, ARNT, and ARNT2 mRNAs were widely distributed throughout the brain and brainstem. There was no neuroanatomic evidence that AhR is preferentially colocalized with ARNT or ARNT2. However, ARNT2, unlike ARNT expression, was relatively high in most regions. The most noteworthy regions in which we found AhR, ARNT, and ARNT2 mRNA were several hypothalamic and brainstem regions involved in the regulation of appetite and circadian rhythms, functions that are disrupted by dioxin exposure. These regions included the arcuate nucleus (Arc), ventromedial hypothalamus (VMH), paraventricular nucleus (PVN), suprachiasmatic nucleus (SCN), nucleus of the solitary tract (NTS), and the dorsal and median raphe nuclei. This neuroanatomic information provides important clues as to the sites and mechanisms underlying the previously unexplained effects of dioxins in the central nervous system. PMID- 11054705 TI - Time course of degenerative alterations in nigral dopaminergic neurons following a 6-hydroxydopamine lesion. AB - The neurotoxin 6-hydroxydopamine (6-OHDA) has been used extensively in animal models of Parkinson's disease. Typically, rodents develop severe unilateral movement deficiencies coupled with apomorphine-induced rotation behavior at least 1 week after an ipsilateral 6-OHDA lesion of the nigrostriatal dopamine (DA) system. The short-term morphological effects of 6-OHDA have not been determined in detail, however, and the exact process by which neurons die has not been elucidated. Thus, novel degenerative markers were used to determine the temporal pattern of acute phenotypic and degenerative alterations following a unilateral 6 OHDA injection into the medial forebrain bundle of adult rats. 6-Hydroxydopamine administration resulted in an increase in terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) staining as early as 6 hours postlesion. Staining for FluoroJade, a marker of neuronal degeneration, was evident at all time points examined but was maximal at 48 hours. Loss of tyrosine hydroxylase (TH) immunoreactivity began in axons at 6 hours, and progressed to cell bodies at later time points postlesion. Morphological examination of these neurons supported the conclusion of their death via apoptosis. Thus, whereas behavioral manifestations typically become evident 1 week or more following a 6-OHDA lesion, it is evident that nigral cell degeneration begins much earlier. This suggests multiple therapeutic possibilities, including the prevention of apoptosis, in affected neurons. PMID- 11054706 TI - Immunocytochemical localization of glutamate receptor subunits in the brain stem and cerebellum of the turtle Chrysemys picta. AB - The regional distribution of ionotropic (AMPA and NMDA) and metabotropic (mGluR1alpha) glutamate receptor subunits was examined in the brain stem and cerebellum of the pond turtle, Chrysemys picta, by using immunocytochemistry and light microscopy. Subunit-specific antibodies that recognize NMDAR1, GluR1, GluR4, and mGluR1alpha were used to identify immunoreactive nuclei in the brain stem and cerebellum. Considerable immunoreactivity in the turtle brain stem and cerebellum was observed with regional differences occurring primarily in the intensity of staining with the antibodies. The red nucleus, lateral reticular nucleus and cerebellum labeled intensely for NMDAR1 and moderately for GluR1. The cerebellum also labeled strongly for mGluR1alpha. All of the cranial nerve nuclei labeled intensely for NMDAR1 and to varying degrees for GluR1, GluR4, and mGluR1alpha. Counterstaining revealed the presence of neuronal somata where there were no immunoreactive neurons in individual nuclei. This finding suggests that there are subpopulations of immunoreactive neurons within a given nucleus that bear different glutamate receptor subunit compositions. The results suggest that the glutamate receptor subunit distribution in the brain stem and cerebellum of turtles is similar to that reported for rats. Additionally, there is considerable colocalization of NMDA and AMPA receptors as revealed by light microscopy. These results have implications for the organization of neural circuits that control motor behavior in turtles, and, generally, for the function of brain stem and cerebellar neural circuits in vertebrates. PMID- 11054707 TI - Gradients of ephrin-A2 and ephrin-A5b mRNA during retinotopic regeneration of the optic projection in adult zebrafish. AB - Regeneration of optic axons in the continuously growing optic system of adult zebrafish was analyzed by anterograde tracing and correlated with the mRNA expression patterns of the recognition molecules ephrin-A2 and ephrin-A5b in retinal targets. The optic tectum and diencephalic targets are all reinnervated after a lesion. However, the rate of erroneous pathway choices was increased at the chiasm and the bifurcation between the ventral and dorsal brachium of the optic tract compared to unlesioned animals. Tracer application to different retinal positions revealed retinotopic reinnervation of the tectum within 4 weeks after the lesion. In situ hybridization analysis indicated the presence of rostral-low to caudal-high gradients of ephrin-A2 and ephrin-A5b mRNAs in unlesioned control tecta and after a unilateral optic nerve lesion. By contrast, the parvocellular superficial pretectal nucleus showed retinotopic organization of optic fibers but no detectable expression of ephrin-A2 and ephrin-A5b mRNAs. However, a row of cells delineating the terminal field of optic fibers in the dorsal part of the periventricular pretectal nucleus was intensely labeled for ephrin-A5b mRNA and may thus provide a stop signal for ingrowing axons. Ephrin-A2 and ephrin-A5b mRNAs were not detectable in the adult retina, despite their prominent expression during development. Thus, given a complementary receptor system in retinal ganglion cells, expression of ephrin-A2 and ephrin-A5b in primary targets of optic fibers in adult zebrafish may contribute to guidance of optic axons that are continuously added to the adult projection and of regenerating axons after optic nerve lesion. PMID- 11054708 TI - The enigmatic inflammatory pseudotumours: the current state of our understanding, or misunderstanding. AB - The inflammatory pseudotumour is a bona fide tumour in the sense of a mass lesion, which is known to present in virtually every anatomic region and organ from the central nervous system to the gastrointestinal tract. A fundamental question about pathogenesis is whether the inflammatory pseudotumour is a pseudo or true neoplasm. There is evidence to support the argument that some of these fibroinflammatory masses are infection-associated and are often characterized by a proliferation of spindled histiocytes and/or dendritic cells, in contrast to a myofibroblastic proliferation in the other inflammatory pseudotumour, also known as the inflammatory myofibroblastic tumour. PMID- 11054709 TI - Proteomics: a new approach to the study of disease. AB - The global analysis of cellular proteins has recently been termed proteomics and is a key area of research that is developing in the post-genome era. Proteomics uses a combination of sophisticated techniques including two-dimensional (2D) gel electrophoresis, image analysis, mass spectrometry, amino acid sequencing, and bio-informatics to resolve comprehensively, to quantify, and to characterize proteins. The application of proteomics provides major opportunities to elucidate disease mechanisms and to identify new diagnostic markers and therapeutic targets. This review aims to explain briefly the background to proteomics and then to outline proteomic techniques. Applications to the study of human disease conditions ranging from cancer to infectious diseases are reviewed. Finally, possible future advances are briefly considered, especially those which may lead to faster sample throughput and increased sensitivity for the detection of individual proteins. PMID- 11054710 TI - Intestinal inflammatory pseudotumour with regional lymph node involvement: identification of a new bacterium as the aetiological agent. AB - Inflammatory pseudotumours are the morphological expression of diverse processes such as reactive/reparative, infective, and neoplastic. This paper reports an example of intestinal inflammatory pseudotumour, with identification of a newly characterized bacterium in the lesion. The patient presented with intestinal obstruction. Laparotomy revealed a tumour in the terminal ileum causing stricture, and multiple enlarged regional lymph nodes. Histologically, the tumour and lymph nodes were composed of plump spindle cells disposed in a vague storiform pattern, and associated with lymphocytes and plasma cells. Immunohistochemical studies showed that most of the spindle cells were histiocytes (CD68 positive), prompting a search for a bacterial aetiology, akin to mycobacterial spindle cell pseudotumour. All histochemical stains for micro organisms were unrewarding. Ultrastructural studies, however, revealed abundant bacteria within the spindle histiocytes. Polymerase chain reaction, using conserved oligonucleotide primers complementary to the 16S rRNA genes of eubacteria, was employed to amplify 16S rRNA gene fragments directly from the involved lymph node tissue. Phylogenetic analysis of the amplified DNA sequences revealed an organism with 99% sequence conformity to Pseudomonas veronii, a bacterium which has hitherto not been implicated in human infection. The importance of searching for an infective agent in inflammatory pseudotumour in the appropriate setting is re-emphasized. PMID- 11054711 TI - Metallothionein in human gastrointestinal cancer. AB - Metallothionein (MT) is a small thiol-rich metalloprotein with antioxidant properties, involved in tumour pathophysiology and therapy resistance. In order to assess the contribution of MT in gastrointestinal carcinogenesis, this study examined both the MT content by radioimmunoassay and the MT localization by immunohistochemistry in pairs of neoplastic and normal-appearing human gastrointestinal tissues. In addition, the relationship between MT expression and major clinicopathological parameters was assessed. The MT concentration of gastric carcinomas and of colorectal adenomas, carcinomas, and liver metastases was found to be significantly lower than that of corresponding normal-appearing tissue. A relatively high MT content, however, was found to be associated with the villous character of colorectal adenomas and with the Dukes' stage of colorectal carcinomas, indicating a relationship between MT level and malignant potential. Immunohistochemical evaluation showed a fairly good correlation with these quantitative data. MT was found to be expressed at a low level and in a patchy pattern in the gastrointestinal neoplastic and metastatic tissues, whereas in normal-appearing gastrointestinal mucosa MT was uniformly distributed in the cytoplasm and/or nucleus of apical cells. Although in the gastric cancer patients no association was found between the MT concentration and the clinicopathological parameters, the strong MT expression in areas with intestinal metaplasia, known to have neoplastic potential, further points to a relationship between this antioxidant metalloprotein and the malignant character of cells. Gastrointestinal neoplasms are apparently accompanied by a low level and decreased expression of MT, but those with a relatively high level seem to have an increased malignant potential. Further studies will be required to determine the clinical relevance of these observations. PMID- 11054712 TI - Helicobacter pylori-related and -non-related gastric cancers do not differ with respect to chromosomal aberrations. AB - Gastric carcinogenesis is strongly associated with Helicobacter pylori infection, but the underlying genetic mechanisms are largely unknown. The aim of this study was to correlate chromosomal aberrations in gastric cancer to H. pylori status and its different strains, as well as to histological type and other clinico pathological variables. DNA from 46 gastric cancers (male/female 35/11, age 27-85 years) was extracted from formaldehyde-fixed, paraffin-embedded material and tested for chromosomal gains and losses by comparative genomic hybridization (CGH). Chromosomal aberrations with frequencies of 20% or higher were considered to be non-random changes associated with gastric cancer. The mean number of chromosomal events per tumour was 9.7 (range 0-27), with a mean of 3.2 gains (range 0-16) and 6.5 losses (range 0-15). Gains were most frequently found at chromosomes 8q and 13q (24% and 26%, respectively). Losses were predominantly found on chromosome arms 2q, 9p, 12q, 14q, 15q, 16p, 16q, 17p, 17q, 19p, 19q, and 22q (22%, 30%, 43%, 22%, 33%, 50%, 28%, 50%, 39%, 33%, 39%, and 37%, respectively). Common regions of overlap narrowed down to 2q11-14, 8q23, 9p21, 12q24, 13q21-22, 14q24 and 15q11-15. The mean number of gains was higher in tumours with metastases than in localized tumours (4.1 vs. 1.9, p=0.04). Tumours with a loss at 17p showed a higher number of losses than tumours without a 17p loss (9. 5 vs. 4.7 on average, p<0.001). Neither H. pylori status (+, n=25; -, n=21) nor H. pylori strain was correlated to the total number of events or to any specific chromosomal aberration, nor were there differences between intestinal (n=30) and diffuse (n=15) cancers or any other clinico-pathological variable tested. In conclusion, a complex of chromosomal aberrations is involved in gastric cancer, but their pattern does not depend on H. pylori status or strain, nor on the histological type of the tumour. The exact biological meaning of these aberrations in carcinogenesis needs further clarification. PMID- 11054713 TI - Marked genetic similarities between hepatitis B virus-positive and hepatitis C virus-positive hepatocellular carcinomas. AB - Hepatocellular carcinoma (HCC) is one of the most common neoplasms worldwide. Well-established risk factors include infections with two very different viruses: the DNA virus causing hepatitis B (HBV) and the RNA virus inducing hepatitis C (HCV). In order to determine whether genetic differences exist between HBV- and HCV-induced HCC, 41 HCC samples of known vival status were examined by comparative genomic hybridization (CGH). The analysis revealed frequent deletions of 1p (24%), 4q (39%), 6q (41%), 8p (44%), 9p (24%), 11q (24%), 12q (22%), and 13q (39%), as well as common gains of 1q (46%), 6p+ (20%), 8q+ (41%), 11q (27%), and 17q+ (37%). There was no significant difference in the number and type of chromosomal imbalances between 25 HCV- and 16 HBV-infected tumours. This is consistent with models suggesting that HBV and HCV cause cancer through non specific inflammatory and regenerative processes, rather than through virus specific interactions with defined target genes. Chromosomal imbalances were also unrelated to the grade and stage of HCC. This may suggest that most gross genomic alterations occur early during HCC development and that further progression of these tumours may be associated with other types of genetic changes, not detectable by CGH. In summary, these data show that characteristic gross genomic changes occur in HCC, but these alterations at present do not appear to have diagnostic or prognostic applications. PMID- 11054714 TI - Lumican and decorin are differentially expressed in human breast carcinoma. AB - Previous studies have shown that lumican is expressed and increased in the stroma of breast tumours. Lumican expression has now been examined relative to other members of the small leucine-rich proteoglycan gene family in normal and neoplastic breast tissues, to begin to determine its role in breast tumour progression. Western blot study showed that lumican protein is highly abundant relative to decorin, while biglycan and fibromodulin are only detected occasionally in breast tissues (n=15 cases). Further analysis of lumican and decorin expression performed in matched normal and tumour tissues by in situ hybridization showed that both mRNAs were expressed by similar fibroblast-like cells adjacent to epithelium. However, lumican mRNA expression was significantly increased in tumours (n=34, p<0.0001), while decorin mRNA was decreased (p=0.0002) in neoplastic relative to adjacent normal stroma. This was accompanied by a significant increase in lumican protein (n=12, p=0.0122), but not decorin. Further evidence of altered lumican expression in breast cancer was manifested by discordance between lumican mRNA and protein localization in some regions of tumours but not in adjacent morphologically normal tissues. It is concluded that lumican is the most abundant of these proteoglycans in breast tumours and that lumican and decorin are inversely regulated in association with breast tumourigenesis. PMID- 11054715 TI - Reduced CD44 standard expression is associated with tumour recurrence and unfavourable outcome in differentiated thyroid carcinoma. AB - CD44 was detected with an antibody recognizing all forms of CD44 (CD44 standard) and others specific for its v3 and v6 variant isoforms; their prognostic value was evaluated in 213 patients with differentiated thyroid carcinoma (DTC). The staining patterns of CD44 standard (s) and CD44v6 in tumour tissue were quite similar, 176 cases (83%) being highly positive for CD44s and 153 cases (72%) for CD44v6. Only 18 (9%) tumours showed high expression of CD44v3. Papillary carcinomas were significantly more often high expressors of CD44s and CD44v6 than follicular carcinomas (p<0.001 for both). Age older than 60 years, distant metastases, and advanced pTNM stage were related to loss of expression of CD44s (p<0.001, p=0.021, and p=0.003, respectively). Tumour recurrence and cancer related mortality were related to the reduced level of CD44s (p=0.049 and p=0.042). CD44v3 did not associate with any of the clinicopathological factors. In univariate analysis, CD44s was the only significant prognostic factor for disease-free survival (p=0.0488). In multivariate analysis, CD44s and thyroglobulin level were significant prognostic factors for disease-free survival (p=0.040 and p<0.001, respectively). The reduced level of CD44s in DTC patients seems to be an independent prognostic factor for unfavourable disease outcome. PMID- 11054716 TI - Prediction of a mismatch repair gene defect by microsatellite instability and immunohistochemical analysis in endometrial tumours from HNPCC patients. AB - Instability of microsatellite repeat sequences has been observed in colorectal carcinomas and in extracolonic malignancies, predominantly endometrial tumours, occurring in the context of hereditary non-polyposis colorectal cancer (HNPCC). Microsatellite instability (MSI) as a feature of human DNA mismatch repair (MMR) driven tumourigenesis of the uterine mucosa has been studied primarily in sporadic tumours showing predominantly somatic hypermethylation of MLH1. The present study shows that all endometrial carcinomas (n=12) from carriers of MLH1 and MSH2 germline mutations demonstrate an MSI-high phenotype involving all types of repeat markers, while in endometrial carcinomas from MSH6 mutation carriers, only 36% (4 out of 11) demonstrate an MSI-high phenotype. Interestingly, an MSI high phenotype was found in endometrial hyperplasias from MSH2 mutation carriers, in contrast to hyperplasias from MLH1 mutation carriers, which exhibited an MSI stable phenotype. Instability of only mononucleotide repeat markers was found in both endometrial carcinomas and hyperplasias from MSH6 mutation carriers. In 29 out of 31 (94%) endometrial tumour foci, combined MSI and immunohistochemical analysis of MLH1, MSH2, and MSH6 could predict the identified germline mutation. The observation of MSI in endometrial hyperplasia and of altered protein staining for the MMR genes supports the idea that inactivation of MMR genes is an early event in endometrial tumourigenesis. A correlation was found between the variation in the extent and level of MSI and the age of onset of carcinoma, suggesting differences in the rate of tumour progression. A high frequency of MSI in hyperplasias, found only in MSH2 mutation carriers, might indicate a more rapid tumour progression, correlating with an earlier age of onset of carcinoma. The present study indicates that assessment of altered protein staining combined with MSI analysis of endometrial tumours might direct the mutational analysis of MMR genes. PMID- 11054717 TI - Assessment of cellular expression of parathyroid hormone-related protein mRNA and protein in multiple myeloma. AB - The capacity of multiple myeloma cells to generate parathyroid hormone-related protein (PTHrP) has been examined by in situ assessment of PTHrP mRNA and PTHrP protein in myeloma cells of patients in whom the disease was associated with the development of hypercalcaemia. The presence of PTHrP mRNA was evaluated by in situ hybridization using an antisense riboprobe, and PTHrP by immunohistochemistry using a monoclonal antibody, in archival bone marrow trephine specimens from 17 patients. PTHrP mRNA was detected in myeloma cells in 16 of the 17 patients, indicating a high frequency of PTHrP gene expression in myeloma cells in these subjects. PTHrP protein was, on the other hand, detected in the myeloma cells of only five of these patients. The impact of the mercury based fixation and decalcification procedure used for processing the bone marrow trephine specimens was assessed to determine the influence of this process on the outcome of the immunohistochemical assay for PTHrP. It was shown that this preparative procedure resulted in a marked reduction of immunohistochemically detectable PTHrP, which provides a possible explanation for the lower frequency of positivity for PTHrP in myeloma cells in the bone marrow specimens. The present findings are consistent with the view that PTHrP can be generated in myeloma cells in vivo, and could contribute to osteolysis and hypercalcaemia, as in patients with cancer. PMID- 11054718 TI - Prognostic value of the preserved expression of the E-cadherin and catenin families of adhesion molecules and of beta-catenin mutations in synovial sarcoma. AB - This study addresses the immunohistochemical expression of the E-cadherin and catenin families and mutations of the beta-catenin gene detected by PCR-SSCP in synovial sarcoma. Immunohistochemical analysis was performed for 72 cases, with follow-up data available on 62. The prognostic value of the expression of these proteins was evaluated. Reduced immunoreactivity for E-cadherin and alpha-catenin was significantly correlated with a poor survival rate (p=0.0040 and 0.0053, respectively). According to multivariate analysis, low AJC stage (stages I and II: p<0.0001), the preservation of alpha-catenin expression (p=0.0001), and a low necrotic rate (<50%: p=0.0139) were independent favourable prognostic factors. Widespread aberrant staining of beta-catenin protein within cytoplasm and/or nuclei was observed in 28 cases (38.9%) and was significantly correlated with poor survival (p=0.0122). In addition, there was a trend towards a correlation between widespread aberrant staining of beta-catenin and the MIB-1 labelling index (p=0.0535). Mutational analysis of exon 3 of the beta-catenin gene was performed for 49 cases. Nucleotide sequencing analysis revealed that four (8.2%) contained point mutations (three in codon 32, GAC to TAC; one in codon 37, TCT to TTT). Survival data were available for three out of four cases with beta-catenin mutations; two of these patients died within 1 year (died of disease at 6 and 11 months, respectively). These results suggest that E-cadherin and alpha-catenin undertake important roles as intercellular adhesion molecules; their preserved expression is associated with a better overall survival rate in synovial sarcoma and may have prognostic value. Abnormal levels of beta-catenin, with or without mutation, could contribute to the development and progression of synovial sarcoma, through increasing the proliferative activity of the tumour cells. PMID- 11054719 TI - An automated machine vision system for the histological grading of cervical intraepithelial neoplasia (CIN). AB - The histological grading of cervical intraepithelial neoplasia (CIN) remains subjective, resulting in inter- and intra-observer variation and poor reproducibility in the grading of cervical lesions. This study has attempted to develop an objective grading system using automated machine vision. The architectural features of cervical squamous epithelium are quantitatively analysed using a combination of computerized digital image processing and Delaunay triangulation analysis; 230 images digitally captured from cases previously classified by a gynaecological pathologist included normal cervical squamous epithelium (n=30), koilocytosis (n=46), CIN 1 (n=52), CIN 2 (n=56), and CIN 3 (n=46). Intra- and inter-observer variation had kappa values of 0.502 and 0.415, respectively. A machine vision system was developed in KS400 macro programming language to segment and mark the centres of all nuclei within the epithelium. By object-oriented analysis of image components, the positional information of nuclei was used to construct a Delaunay triangulation mesh. Each mesh was analysed to compute triangle dimensions including the mean triangle area, the mean triangle edge length, and the number of triangles per unit area, giving an individual quantitative profile of measurements for each case. Discriminant analysis of the geometric data revealed the significant discriminatory variables from which a classification score was derived. The scoring system distinguished between normal and CIN 3 in 98.7% of cases and between koilocytosis and CIN 1 in 76.5% of cases, but only 62.3% of the CIN cases were classified into the correct group, with the CIN 2 group showing the highest rate of misclassification. Graphical plots of triangulation data demonstrated the continuum of morphological change from normal squamous epithelium to the highest grade of CIN, with overlapping of the groups originally defined by the pathologists. This study shows that automated location of nuclei in cervical biopsies using computerized image analysis is possible. Analysis of positional information enables quantitative evaluation of architectural features in CIN using Delaunay triangulation meshes, which is effective in the objective classification of CIN. This demonstrates the future potential of automated machine vision systems in diagnostic histopathology. PMID- 11054720 TI - In vitro-generated stem cell leukaemia showing altered cell cycle progression with distinct signalling of the tyrosine-phosphorylated rasGAP-associated p62(dok) protein. AB - In an attempt to gain more insight into the events of leukaemic transformation, a cell line overexpressing MHC class II (DR) was generated by transfecting an early CD34-negative haematopoietic progenitor stem cell line with the appropriate constructs. The stable transfection with genes for DR antigens leads to cellular transformation. The DR(+) transformed cell clones express a tyrosine phosphorylated DR heterodimer and show a significantly different morphology. DR(+) clones present the morphology of an immature myeloid neoplasia expressing alpha-naphthyl-acetate-esterase (ANAE), but neither myeloperoxidase nor CD34. While D064 cells predominately grow adherent as fibroblast-like cells, the DR(+) clones display a decrease in adherent growth. Although both cell lines express similar amounts of the interleukin-6 (IL-6) signal transducer gp130, the DR transfected cells still show activation of STAT factors by IL-6, whereas D064 cells do not. Although the transformed clones present acceleration of cell-cycle transition and growth, the G(0)/G(1) progression inhibitor p27(kip-1) is up regulated, while the expression of proteins involved in the S/G(2) phase transition, such as cyclin B and cdc2 (p34), is suppressed. Instead cyclin D3, one of the G(0)/G(1) progression factors, is up-regulated, as well as tyrosine phosphorylated p62(dok), suggesting dysregulation of cell cycle-controlling proteins. In addition, DR(+) leukaemia-like cells also overexpress Bcl-2, while bax expression is suppressed, compared with the wild-type (wt) parental haematopoietic stem cell line. PMID- 11054721 TI - Deep intralobular extension of human hepatic 'progenitor cells' correlates with parenchymal inflammation in chronic viral hepatitis: can 'progenitor cells' migrate? AB - Ductular reaction and putative progenitor cells (or 'progenitor cells'), which are presumed to be the human counterpart of the oval cells in rat liver, have been discerned in various human liver diseases, including chronic viral hepatitis. Since in experimental models of chronic hepatitis the activation of oval cells is correlated with the inflammatory infiltrate, this study investigated whether there is a correlation in chronic viral hepatitis between the number of 'progenitor cells' extending into the lobule and the severity of parenchymal inflammation, on the one hand, and the extent of ductular reaction and the severity of interface hepatitis, on the other hand. Liver biopsies of 55 patients with chronic hepatitis B and/or C were used. The severity of parenchymal inflammation and of interface hepatitis was semiquantitatively graded on a haematoxylin and eosin-stained paraffin section, while the number of 'progenitor cells' and the extent of the ductular reaction were assessed on a serial section stained for cytokeratin (CK) 7. In addition, more extensive phenotyping of 'progenitor cells' was performed on sections from frozen material from five patients, using antibodies against CK7, CK8, CK18, CK19, chromogranin-A, and the rat oval cell marker OV-6. The number of more centrally located 'progenitor cells' correlated significantly with the severity of the parenchymal inflammation, while the extent of the ductular reaction correlated significantly with the severity of interface hepatitis. These findings suggest that in chronic viral hepatitis, inflammation plays a role in 'progenitor cell' activation and its topography. In cases with moderate and severe lobular inflammation, 'progenitor cells' were strikingly scattered throughout the parenchyma and surrounded by intermediate hepatocyte-like cells, suggesting their migration into the parenchyma and their differentiation towards the hepatocytic lineage. PMID- 11054722 TI - Expression of pulmonary vascular angiotensin-converting enzyme in primary and secondary plexiform pulmonary hypertension. AB - The hypothesis for this study was that increased local expression of vascular angiotensin-converting enzyme (ACE) may contribute to the arterial remodelling which accompanies pulmonary hypertension, since angiotensin II (ANG II) is an important mediator of pulmonary vascular cell growth. The expression of ACE was studied by immunohistochemistry in paraffin-embedded lung sections from adults undergoing heart-lung transplantation for severe primary (n=6) and secondary (n=7) pulmonary arterial hypertension (PH), compared with age-matched controls (n=11). An antigen retrieval technique was used prior to incubating sections with the anti-ACE monoclonal antibody, CG2, or the endothelial marker, monoclonal anti CD31. In control lungs, the highest level of ACE immunostaining was seen in the alveolar capillary endothelium, with less intense staining in small intra-acinar pulmonary arteries and relatively little staining in larger preacinar arteries. ACE immunostaining was virtually absent in lymphatics and veins. In both primary and secondary PH, there was an increase in ACE immunostaining in the endothelium of intra-acinar peripheral pulmonary arteries compared with control lungs, extending to the level of alveolar ducts, as confirmed by semi-quantitative analysis. The increase in endothelial ACE expression in the intra-acinar arteries of patients with primary and secondary PH is consistent with the hypothesis that locally increased production of ANG II may contribute to the process of pulmonary vascular remodelling. PMID- 11054723 TI - A light and electron microscopic study of osteogenesis imperfecta bone samples, with reference to collagen chemistry and clinical phenotype. AB - A detailed morphological study was carried out using light and electron microscopy on 36 bone specimens from patients suffering from osteogenesis imperfecta (OI) and 20 age- and site-matched control bone specimens. The findings were grouped into the clinical types of OI according to the Sillence classification. The morphological and ultrastructural alterations observed in OI bone correlate well with clinical severity. Thus, OI type I, the mildest type, showed the least abnormalities in bone ultrastructure. OI type IV closely resembled type I, with only minor abnormalities in the bone cells and osteoid. OI type III showed abnormalities in the structure and distribution of osteoid collagen fibrils, whilst OI type II, the lethal form, revealed many varied abnormalities such as thin cortical bone, sparse trabecular bone, increased numbers of osteoclasts and osteocytes, thin osteoid with thin collagen fibrils, and patchy mineralization. PMID- 11054724 TI - Growth of nerve fibres into murine peritoneal adhesions. AB - Adhesions in the peritoneal cavity have been implicated in the cause of intestinal obstruction and infertility, but their role in the aetiology of chronic pelvic pain is unclear. Nerves have been demonstrated in human pelvic adhesions, but the presence of pain-conducting fibres has not been established. The purpose of this study was to use an animal model to examine the growth of nerves during adhesion formation at various times following injury and to characterize the types of fibres present. Adhesions were generated in mice by injuring the surface of the caecum and adjacent abdominal wall, with apposition. At 1-8 weeks post-surgery, adhesions were processed and nerve fibres characterized histologically, immunohistochemically, and ultrastructurally. Peritoneal adhesions had consistently formed by 1 week after surgery and from 2 weeks onwards, all adhesions contained some nerve fibres which were synaptophysin, calcitonin gene-related peptide, and substance P-immunoreactive, and were seen to originate from the caecum. By 4 weeks post-surgery, nerve fibres were found to originate from both the caecum and the abdominal wall, and as demonstrated by acetylcholinesterase histochemistry, many traversed the entire adhesion. Ultrastructural analysis showed both myelinated and non-myelinated nerve fibres within the adhesion. This study provides the first direct evidence for the growth of sensory nerve fibres within abdominal visceral adhesions in a murine model and suggests that there may be nerve fibres involved in the conduction of pain stimuli. PMID- 11054725 TI - Differential expression of laminin chains and anti-laminin autoantibodies in experimental lupus nephritis. AB - Mice with chronic graft-versus-host disease (GvHD) develop a lupus-like disease with severe immune complex glomerulonephritis. Previous studies with this model have shown that anti-laminin autoantibodies are involved in immune complex formation and that glomerular laminin expression alters qualitatively. The present study investigated glomerular laminin chain expression and autoantibody reactivity with matrix antigens during disease development in mice with chronic GvHD, killed before and 6, 8, 10, and 11 weeks after disease induction, using antibodies raised against laminin chain peptides, in immunofluorescence and western blotting studies. Decreased glomerular expression of the laminin beta1 chain, unaltered expression of the laminin beta2 and gamma1 chains, and increased expression of the laminin alpha1 chain and filamin/actin-binding protein 280 (ABP 280) were found during disease progression. Furthermore, 4 weeks after disease induction, autoantibodies appeared which were reactive with laminin alpha1, beta1, beta2, and gamma1 chains, and filamin in rat mesangial cell matrix. Ten weeks after disease induction, autoantibodies reacted with filamin, and beta2 and gamma1 laminin chains. Autoantibodies reacted with laminin chains only and not with other proteins in matrices extracted from glomeruli of normal and diseased mice. Staining with H50, an anti-laminin alpha1 chain/anti-filamin monoclonal autoantibody derived from an MRL/lpr mouse with spontaneous lupus nephritis, confirmed these observations and showed identical anti-laminin/anti-filamin autoantibody reactivity in two different models for lupus nephritis. In summary, differential glomerular expression of laminin chains was found during the development of chronic GvHD. Concomitantly with expression of the laminin alpha1 chain and/or filamin in the glomerulus, anti-laminin alpha1 and/or anti-filamin reactivity was present, pointing towards a role for (neo) antigen expression in the epitope spreading of the immune response. Furthermore, glomerular expression of laminin beta1 decreased in conjunction with decreased presence of anti-laminin beta1 reactivity, presumably due to antigen masking or shedding of immune complexes into the urine. These changes in anti-laminin chain autoantibodies, with concomitant alterations in the glomerular expression of laminin chains, may aggravate progressive immune injury in this model for lupus nephritis. PMID- 11054726 TI - IL-8 mRNA expression in primary malignant melanoma mRNA in situ hybridization: sensitivity, specificity, and evaluation of data. PMID- 11054727 TI - Authors' reply PMID- 11054728 TI - Editorial note PMID- 11054729 TI - Understanding and using the history of social psychology. AB - Authors in this collection offer both critique and contextualist counterpoint to the standard, "official" histories of the field-successive editions of the Handbook of Social Psychology in 1954, 1968, 1985, and 1998. Unlike mainstream histories, the collected studies do not together constitute a seamless chronicle of continual progress for practitioners in a research area seeking social science status, viability, and legitimacy. Rather the authors focus on choice points, crises, and debates (some still ongoing), pay special heed to non-mainstream branches and voices, question numerous assumptions concerning the interrelationships among social psychological methodology, ontology (Danziger; MacMartin & Winston; Stam, Radtke, & Lubek), boundaries (Good), and individualisms (moral, political, and/or methodological). The specific contributions of Floyd and Gordon Allport are discussed from several perspectives as they helped define and shape and write the history of the field (Lubek & Apfelbaum; Parkovnick; Greenwood; Chung), and bridge it to neighboring areas (personality) and disciplines (psychology and sociology) (Nicholson; Barenbaum; Cherry). The constraints, origin myths, insensitivities, and omissions of standard histories are pointed out (Samelson), some partial correctives are advanced, and a more generative role for future historical studies is suggested. PMID- 11054730 TI - Making social psychology experimental: a conceptual history, 1920-1970. AB - The historical emergence of a field devoted to the experimental investigation of effects identified as "social" required a radical break with traditional conceptions of the social. Psychological experimentation was limited to the investigation of effects that were proximal, local, short-term, and decomposable. A viable accommodation to these constraints occurred in the closely related programs of Moede's experimental crowd psychology and Floyd Allport's experimental social psychology. Later, Kurt Lewin attempted to provide a different conceptual foundation for the field by drawing on certain precepts of Gestalt psychology and the philosophy of scientific experimentation developed by Ernst Cassirer. These ideas were poorly understood and were soon replaced by a methodological regime in which a new generation of statistical procedures and experimental design shaped implicit conceptions of the social in social psychological experiments through such procedures as randomization and the additive combination of variables. PMID- 11054731 TI - The rhetoric of experimental social psychology, 1930-1960: from caution to enthusiasm. AB - Between 1930 and 1960, experimentation became the premier form of knowledge generation in social psychology. In journals, texts, and handbooks, experiment was now conceived as the active manipulation of an independent variable, and the sole method for the discovery of "causes." Understanding this change requires further investigation of the fine-grained discursive strategies used to promote experimentation during the 1930s and 1940s. In this paper we use discourse analysis to contrast the cautious rhetoric used by Gardner Murphy and Lois Murphy and the more enthusiastic, unhedged arguments for experimentation employed by Kurt Lewin. We argue that analysis of changes in discourse justifying experimentation can illuminate the processes by which methodological consensus was constructed. PMID- 11054732 TI - Strains in experimental social psychology: a textual anaylsis of the development of experimentation in social psychology. AB - A textual analysis of post-World War II social psychology methodology manuals and handbook chapters on "methods" indicates that the introduction of the experimental method was enforced and gradually strengthened through the use of scientific rhetoric and the minimization of alternative research strategies. As a consequence, by the 1960s experimentation had become such an established identifying feature of psychological social psychology that the acceptability of ideas in the field came to depend largely on the ability of authors to couch them in the language of the experiment. Text writers continually shored up the defenses of scientific legitimacy and denigrated all other types of argument. We explore three sources of tension or strains evident as contradictions in these texts: (1) between a rational experimenter's carefully following prescribed, logic-generated scientific practices and the investigator's artfully or intuitively designing research; (2) between social psychologists' missionary activities of proselytizing the experiment as the primary research method and social psychologists' apologies and insecurities expressed about using experiments; and (3) between the treatment of participants as docile and submissive versus portraying them as underhanded and damaging to the outcome of the research. In addition, we briefly reexamine the strain (4) between sober scientific experimentation and a playful "fun and games" approach to experimentation (Lubek & Stam, 1995). PMID- 11054733 TI - Disciplining social psychology: a case study of boundary relations in the history of the human sciences. AB - This paper explores the disciplinary status of social psychology through an analysis of the history of the boundary relations of psychology, sociology, and social psychology. After outlining some research on the nature of scientific disciplines, on the role of rhetoric in the constitution of disciplines, and on "boundary work," I consider the singular importance of social psychology as a discipline for the analysis of boundary relations, examining its units of analysis and its "disciplining." The boundaries of the disciplines of social psychology were seen as fluid, contingent, local, and contestable, reflecting the thematic preoccupations, disciplinary origins, and meta-theoretical commitments of social psychologists, of the parent disciplines, and of those who represent disciplinary practices. PMID- 11054734 TI - A critical gaze and wistful glance at Handbook histories of social psychology: did the successive accounts by Gordon Allport and successors historiographically succeed? AB - Gordon Allport's account of the development of social psychology in the 1954 Handbook of Social Psychology became, de facto, a standard or official historical reference for researchers and apprentices. His history also provided the field's ontological center point with a definition of social psychology that would become predominant. The revised and updated chapter appeared posthumously in 1968, was then reprinted (lightly edited) in 1985, but was removed from the 1998 Handbook. In 1966, Allport prepared a parallel evaluation of six decades of the history of social psychology, for a conference on graduate education in social psychology. This paper was critical of "elaborate mendacious experimentation" and ended with a plea for an interdisciplinary cross-cultivation. It was rarely cited. Ironically, it was Allport's "official" history, his justificatory Handbook account, that often was used for graduate mentoring rather than the more critical history, specifically written to address issues of graduate education. Other "official" Handbook historical chapters that succeeded Allport's displayed less breadth of geographical and transdisciplinary coverage and offered a shorter temporal, more presentist, and more selective personalist historical perspective. In contrast to more contextualist accounts, these Handbook chapters are constrained in a number of ways that raise questions about the success, functions, and professional consequences of such "official" histories, and who should write them. PMID- 11054735 TI - Contextualizing Floyd Allports's Social Psychology. AB - This paper looks at the program for social psychology presented by Floyd Allport in his Social Psychology of 1924. It contextualizes Allport's program in terms of intellectual currents of the time and the views of his teachers at Harvard University, specifically the philosopher Ralph Barton Perry and the psychologists Edwin B. Holt and Hugo Munsterberg. Finally, the paper analyzes responses to Allport's program at the time and later, retrospective responses. PMID- 11054736 TI - Individualism and the social in early American social psychology. AB - In this paper an attempt is made to specify the original conception of the social dimensions of cognition, emotion and behavior-and of a distinctively social psychology-that was held by early American social psychologists, but abandoned by later generations of social psychologists committed to Floyd Allport's individualistic experimental program. Two influential forms of "individualism" in the work of Floyd Allport are distinguished and detailed. PMID- 11054737 TI - The compatibility of two generations of American social psychologists. AB - This paper examines Greenwood's (2000) evidence for incompatibility between the early and later American social psychologists on the social conception of cognition, emotion, and behavior. The notion of the autonomy of the individual may offer the key to finding a degree of compatibility between them. Both generations, I argue, fundamentally accept the notion of individual persons as autonomous agents who are able to decide and choose to act and, hence, be responsible for their actions. Philosophical analysis can perhaps inform historians of social psychology on how carefully and critically to reexamine evidence for traditional claims of generational, paradigmatic, and/or foundational splits. PMID- 11054738 TI - "A coherent datum of perception": Gordon Allport, Floyd Allport, and the politics of "personality". AB - This paper examines Floyd and Gordon Allport's early work on "personality" psychology. In the early 1920s, personality was an unorthodox topic, and for the Allports it initially served as an intellectual and personal bond. Floyd proposed the subject to his brother as a dissertation topic, and the two worked closely on developing personality tests. By 1924, however, "personality" had become the site of a dispute between the two brothers over the intellectual and methodological character of American psychology. The present study examines the origins of this dispute, while gauging the personal and professional ramifications of the dispute. On a larger level, this essay explores the role and meaning of "personality" in the academic culture of 1920s America. PMID- 11054739 TI - How social was personality? The Allports' "connection" of social and personality psychology. AB - This paper investigates three conflicting reconstructions of the historical relationship between personality and social psychology and addresses questions they raise regarding the subdisciplinary status of personality in the 1920s and the way in which the field gradually emerged as a separate area of psychology. Contesting claims that Floyd Allport first connected social psychology to a separate "branch" of personality psychology in the 1920s, I argue that he drew upon earlier work of psychologists and sociologists who treated personality as a central topic of social psychology. I compare Floyd Allport's views with those of Gordon Allport, who endeavored to establish personality as a separate subdiscipline. PMID- 11054740 TI - The nature of The Nature of Prejudice. AB - This paper attempts to establish the historical context for the development and publication of Gordon Allport's text, The Nature of Prejudice, and by so doing illustrate the importance of historicizing psychological social psychology. The Nature of Prejudice was, in part, the cumulative result of a decade of Gordon Allport's classroom teaching in a new interdisciplinary unit at Harvard, the Department of Social Relations. This paper chronicles key elements of Allport's course -"Prejudice and Intergroup Conflict"-from 1944 to the mid-1950s. PMID- 11054741 TI - Whig and anti-whig histories- and other curiosities of social psychology. AB - In successive editions of the Handbook of Social Psychology (Lindzey, 1954), the focus of the history of the field shifted from the substantive ideas of nineteenth-century thinkers to the successful emergence of a psychological experimental social psychology in the twentieth. Countering this whiggish account, the dominant themes in the present issue involve attempts to portray two parallel paradigm shifts: from a "social" to an "asocial" social psychology, and from a broad-ranging theoretical-philosophical subject to a narrow experimental (psychological) science-changes initiated by Floyd Allport. But such a formulation may be called into question as another version of retrospective history-with inverted, anti-Whig valuations. PMID- 11054742 TI - Briefly noted PMID- 11054743 TI - News and notes PMID- 11054744 TI - Functional and clinical significance of skeletal muscle architecture. AB - Skeletal muscle architecture is the structural property of whole muscles that dominates their function. This review describes the basic architectural properties of human upper and lower extremity muscles. The designs of various muscle groups in humans and other species are analyzed from the point of view of optimizing function. Muscle fiber arrangement and motor unit arrangement is discussed in terms of the control of movement. Finally, the ability of muscles to change their architecture in response to immobilization, eccentric exercise, and surgical tendon transfer is reviewed. Future integrative physiological studies will provide insights into the mechanisms by which such adaptations occur. It is likely that muscle fibers transduce both stress and strain and respond by modifying sarcomere number in a way more suited to the new biomechanical environment. PMID- 11054745 TI - Physiologic basis of potentials recorded in electromyography. AB - The extracellularly recorded configuration of a single muscle fiber discharge is generally appreciated to be triphasic with an initially positive deflection. However, careful attention to waveform appearance during the electrodiagnostic medicine examination reveals that both innervated and denervated muscle waveforms may display a pantheon of configurations. Further, despite the fact that innervated and denervated single muscle fiber discharges arise from distinctly different intracellular action potential (IAP) configurations, their extracellularly recorded waveforms can appear quite similar, leading to potential misidentification and, hence, the possibility of an erroneous diagnostic conclusion. The least appreciated, but nevertheless critical, aspect of explanations for muscle waveform configurations is the relationship between the muscle fiber and recording electrode. Additionally, it is important to appreciate both the near-field and far-field aspects of single fiber and compound muscle action potentials. In this review, the leading/trailing dipole model is used to explain muscle waveform configurations in both innervated and denervated tissues. PMID- 11054746 TI - Increased lysosome-related proteins in the skeletal muscles of distal myopathy with rimmed vacuoles. AB - Investigators have speculated that the degenerative process in distal myopathy with rimmed vacuoles (DMRV) mainly involves the lysosomal system. To investigate possible protein abnormalities related to intracellular lysosomal proteolytic pathways in DMRV-affected muscles, we performed immunohistochemical analyses of certain proteins in muscle biopsy specimens obtained from patients with various neuromuscular diseases, including DMRV, muscular dystrophy, polymyositis, and amyotrophic lateral sclerosis, and in normal human muscles specimens. Immunohistochemically, most muscle fibers in normal control specimens showed little or no reaction for clathrin and alpha- and gamma-subunits of adaptin constituted adaptin proteins (AP)-1 and AP-2, respectively. Abnormal increases in these proteins were demonstrated mainly in the cytoplasm of atrophic fibers or in necrotic fibers in all diseased specimens. Particularly in DMRV-affected muscles, alpha- and gamma-adaptins were often observed inside or on the rims of vacuoles and in the cytoplasm of vacuolated fibers. Abnormal increases in Golgi-zone protein were also demonstrated in DMRV muscles. The rims of rimmed vacuoles were negative for kinectin, an endoplasmic reticulum-binding protein. Positive staining for both proteins, however, was sometimes seen inside the vacuoles in DMRV-affected fibers. These results suggest increased endocytosis at the plasma membrane as well as secretion involving transport from the trans-Golgi network of the Golgi apparatus in DMRV. Accumulation of various lysosome-related proteins within the rimmed vacuoles indicates at least some of these vacuoles may be autolysosomes. PMID- 11054747 TI - Maximal bite force and surface EMG in patients with myasthenia gravis. AB - Masticatory muscle strength was quantified in patients with bulbar myasthenia gravis and compared with that of patients with ocular myasthenia gravis, patients in clinical remission (whether or not pharmacological) who previously suffered from bulbar myasthenia gravis, and healthy subjects. Maximal bite force and maximal activity of the masseter and temporalis muscles and of the submental muscle complex were measured. Bite force was decreased in the patients with bulbar myasthenia gravis, but was normal in the patients in the clinical remission group and in the ocular group. These findings were consistent with the results of electromyographic data. Although subjective reports of masticatory muscle weakness provide valuable information, quantitative measurements provide more information about the degree of muscle weakness of individual muscles. This is especially important for longitudinal evaluation of therapy in individual patients and for pharmacotherapeutic research. PMID- 11054748 TI - Leukemia inhibitory factor ameliorates muscle fiber degeneration in the mdx mouse. AB - Although the muscles of the mdx mouse lack dystrophin, the protein absent in muscles of humans affected with Duchenne muscular dystrophy (DMD), the only mdx muscle to degenerate in a manner similar to those of DMD boys is the diaphragm. We have previously shown that leukemia inhibitory factor (LIF) is a trauma factor that enhances muscle repair in vivo and, when applied exogenously, increases the fiber size of mdx skeletal muscle. Furthermore, we developed a controlled release device for LIF based on a calcium alginate rod (release rate about 0.5% per day). These rods were sutured to the abdominal surface of the hemidiaphragm of mdx mice 3 months old. At age 6 months the mice were killed and the diaphragm muscles fixed and sectioned. The sections showed obvious muscle degeneration at 3 months of age in mdx mouse diaphragms and further degeneration at 6 months in saline perfused muscle. Hemidiaphragm muscles continuously exposed to LIF over the same period contained more normal myofibers, larger regenerated fibers, and less adipose tissue and other non-contractile tissue. Morphometric analysis of the diaphragm sections was carried out. The LIF-treated animals showed a significant increase in fiber number and size compared to saline rod controls. The amount of nonmuscle (connective tissue and adipose tissue) was significantly reduced and the maximum force-producing capacity of isolated diaphragm muscle strips was higher in LIF-treated mice. The results demonstrate that LIF treatment ameliorates the dystrophic abnormalities in mdx mouse diaphragm. PMID- 11054749 TI - Measurement of central activation failure of the quadriceps femoris in healthy adults. AB - The purpose of this investigation was to describe the relationship between the central activation ratio (CAR) and the percent maximum voluntary effort (% MVE) during isometric quadriceps femoris contractions. Twenty-one healthy, young adults participated in three test sessions. During each session, one of three train types was tested: a 100-HZ 120-ms train, a 100-HZ 250-ms train, or a 50-HZ 500-ms train. Subjects were seated on a force dynamometer and stabilized to perform a 3-5-s isometric knee extension at MVE. Force targets were set at 25, 50, 75, and 100% of the MVE. With 5 min rest between efforts, subjects produced forces at the specified target levels. When each target was reached, the test train was delivered to quantify the amount of central activation. There were no significant differences in CARs across train types during maximal efforts, but during submaximal efforts at 25 and 50%, the 100-HZ 250-ms and 50-HZ 500-ms trains produced significantly lower CARs than the 100-HZ 120-ms train. The relationship between the CAR and the %MVE was curvilinear and best described by a second-order polynomial for all three train types. If tests of central activation are going to be used clinically, it is important to know the relationship between the CAR and voluntary effort; however, further study will be required to extend these results to specific patient populations. PMID- 11054750 TI - Enlarged median nerve in idiopathic carpal tunnel syndrome. AB - To delineate change in median nerve size in carpal tunnel syndrome (CTS), we compared ultrasonograms of nerve cross-sections from patients (201 wrists of 125 women with idiopathic CTS) and controls (200 dominant wrists of 200 women). Major and minor axes, cross-sectional area, and the flattening ratio (major axis/minor axis) were measured at four levels: 1, distal edge of the flexor retinaculum; 2, center of the hook of the hamate; 3, wrist crease; and 4, distal one-third level of the forearm. Axes and areas were greater in the patients at levels 1, 2, and 3. The flattening ratio was greater in the controls at levels 1 and 2, and in the patients at level 3. When the groups were combined, there were linear correlations between the cross-sectional nerve area and electrophysiologic severity at levels 1, 2, and 3. These findings indicate that idiopathic CTS is characterized by severity-correlated intracarpal enlargement of the median nerve and not by compressive deformation, such as a reduction in the caliber of the nerve. PMID- 11054751 TI - Strength-duration properties and their voltage dependence as measures of a threshold conductance at the node of Ranvier of single motor axons. AB - In a number of clinical studies, measurement of axonal strength-duration properties has been used to provide indirect insight into conductances at the node of Ranvier, particularly persistent Na(+) conductance. However, the specificity of any changes is limited because other factors can affect strength duration behavior. The present study was undertaken to define the relationship between different strength-duration measures at rest and at different membrane potentials, and also to determine the limits within which strength-duration behavior can be used as a measure of nodal conductances. The strength-duration time constant (tau(SD)) and rheobase of 20 single motor units in the flexor carpi ulnaris were calculated from thresholds defined using threshold tracking. "True" rheobase and rheobasic latencies were measured using test stimuli of 100-ms duration. For ten units, the technique of latent addition was used to measure threshold changes directly attributable to nodal conductances, and for six units these were compared with strength-duration properties at different membrane potentials. The data indicate that measurements of tau(SD) and rheobase can provide sensitive indicators of conductances present at the node of Ranvier when membrane potential changes. There is a reciprocal relationship between tau(SD) and rheobase for single motor units at different membrane potentials, and this relationship may allow changes in tau(SD) due to depolarization and demyelination to be differentiated. PMID- 11054752 TI - Muscle fatigue during concentric and eccentric contractions. AB - We compared the contribution of central and peripheral processes to muscle fatigue induced in the ankle dorsiflexor muscles by tests performed during concentric (CON) and eccentric (ECC) conditions. Each fatigue test consisted of five sets of 30 maximum voluntary contractions at a constant speed of 50 degrees /s for a 30 degrees range of motion of the ankle joint. The torque produced by the dorsiflexors and the surface electromyogram (EMG) of the tibialis anterior muscle were recorded during the fatigue tests. Before, during, and after the tests, the compound muscle action potential (M wave) and the contractile properties in response to single and paired electrical stimuli, as well as the interpolated-twitch method and postactivation potentiation (PAP), were recorded during isometric conditions. Compared with ECC contractions, the CON ones resulted in a greater (P < 0.05) loss of force (-31.6% vs. -23.8%) and a decrease in EMG activity (-26.4% vs. -17.5%). This difference was most pronounced during the first four sets of contractions, but was reduced during the last set. Activation was not altered by the tests because neither the interpolated-twitch response nor the ratio of the voluntary EMG to the amplitude of the M wave was changed in the two fatigue tests. Although there was no significant difference in M-wave amplitude between the two tests, changes in the twitch parameters and in the PAP were found to be greater in the CON than ECC contractions. It is concluded that the greater alterations in the contractile properties observed during the CON contractions indicate that intracellular Ca(2+)-controlled excitation-contraction (E-C) coupling processes, possibly associated with a higher energy requirement, are affected to a much greater degree than during ECC contractions. PMID- 11054753 TI - Patch clamp studies of the thr1313met mutant sodium channel causing paramyotonia congenita. AB - Paramyotonia congenita (PC) is an autosomal-dominant disorder due to a point mutation in the adult skeletal muscle Na channel gene. Muscle fibers from PC patients have normal membrane properties at 32 degrees C. At 27 degrees C, they are inexcitable, have increased Na conductance, and have a reduced resting membrane potential of -40 mV. To define the biophysical basis for the muscle membrane abnormalities, we performed patch clamp whole-cell and outside-out single Na channel studies at 22 degrees C on cultured human muscle cells from 4 control patients and 2 sisters with PC and the thr1313met mutant Na channel. The whole-cell studies showed no difference in window currents. Unlike cells transfected with the thr1313met mutant Na channel, the inactivation time constant, tau(h), for PC cells was similar to control cells. For PC recordings containing long-duration single Na channel openings, mean open time was prolonged at -60, -40, and -20 mV. The long-duration Na channel openings occurred randomly with no evidence of modal gating. The number of channel openings, occurrence of late openings, and the prolonged mean open time resulted in a sustained inward Na current at -40 mV. We suggest that the biophysical marker of the thr1313met mutant Na channel is a voltage- and temperature-dependent abnormality in mutant single Na channel behavior. PMID- 11054754 TI - Standardization of anal sphincter electromyography: effect of chronic constipation. AB - Severe chronic constipation has been implicated as a cause of damage to the pelvic floor innervation. The aim of the present study was to examine the role of mild to moderate chronic constipation, a condition more relevant for clinical electromyographers, because this complaint is common in patients sent for evaluation of possible neurogenic dysfunction of lower sacral myotomes. A group of 59 subjects without major uroneurological dysfunction, proctological disorders, or neurological abnormalities participated in the study, which involved concentric needle electromyography of the external anal sphincter (EAS). Motor unit potentials (MUPs; sampled using multi-MUP analysis) and interference pattern (IP, sampled using turn/amplitude analysis) of chronically constipated and control subjects were compared. No effect of chronic constipation on MUP/IP parameters compatible with neurogenic injury was found. Our results suggest that mild chronic constipation does not cause damage to the EAS innervation, and that no separate reference values are needed for this group of subjects. PMID- 11054755 TI - Utility of an EMG mapping study in treating cervical dystonia. AB - Intramuscular injections of botulinum toxin are the cornerstone of treatment for cervical dystonia. Controversy exists regarding the necessity for EMG-guided injections. We compared the clinical examination of four movement disorder specialists to an electromyographic (EMG) mapping study. Clinical predictions of individual muscle involvement were only 59% sensitive and 75% specific. Muscle hypertrophy, shoulder elevation, and dominant head vector did not bolster clinical accuracy. An EMG mapping study facilitates identification of dystonic muscles in cervical dystonia, which may enhance botulinum toxin therapy. PMID- 11054756 TI - Lumbosacral plexopathy in pelvic trauma. AB - We reviewed the electrophysiologic data and the etiology of lumbosacral plexopathy in 22 consecutive patients with pelvic trauma referred for electromyography (EMG). Most (68%) patients had sacral fractures or sacroiliac joint separation, 14% had acetabular fractures, and 9% had femoral fractures. Lumbosacral plexopathy was significantly more common (P = 0.0026) among patients with sacral fractures (incidence of 2.03%) than among the entire population of patients with pelvic and acetabular fractures (overall incidence 0. 7%). Patients with acetabular and femoral fractures may have suffered injury to multiple proximal nerves originating from the plexus rather than injury to the plexus, as confirmed by magnetic resonance imaging (MRI) neurogram in select cases. PMID- 11054757 TI - Motor cortex disinhibition of the unaffected hemisphere after acute stroke. AB - We studied motor cortex excitability in the nonlesioned hemisphere of patients with a large cortical infarction. Patients with a severe hemiparesis due to a stroke were compared with age-matched, healthy controls. Paired transcranial magnetic stimuli were applied over the unaffected hemisphere to investigate intracortical inhibition and facilitation. In the patient group, intracortical inhibition was reduced. We suggest that this disinhibition is due to an impairment of transcallosal fibers and may affect recovery. PMID- 11054758 TI - Peripheral neuropathy associated with acquired immunodeficiency syndrome (AIDS) related Burkitt's lymphoma. AB - Peripheral neuropathy associated with acquired immunodeficiency syndrome (AIDS) related Burkitt's lymphoma usually occurs as a toxic effect of chemotherapeutic agents. Whereas primary peripheral nerve involvement is an unusual complication, we report on a human immunodeficiency virus (HIV)-positive patient with Burkitt's lymphoma and sciatic neuropathy due to compression by a lymphomatous mass. Therapy with radiation and chemotherapy was followed by clinical and radiological improvement, but recurrent neurological deficits in a similar distribution occurred later from lymphomatous meningitis. PMID- 11054759 TI - Chronic multiple paraneoplastic syndromes. AB - A patient presented with symptoms of limbic and brainstem encephalitis, motor and sensory neuronopathy, cerebellar dysfunction, and highly positive anti-Hu antibodies. He also harbored P/Q-type calcium channel antibodies and manifested the Lambert-Eaton myasthenic syndrome (LEMS). Small-cell lung cancer was found, and he received both antineoplastic therapy and intravenous immunoglobulin (IVIg). Remission of the malignancy was achieved. Although the anti-Hu-related manifestations improved after therapy, LEMS has persisted, leading to IVIg dependency. PMID- 11054760 TI - Cocaine-induced hypokalemic paralysis. PMID- 11054761 TI - Combined biochemical and molecular diagnosis in blood of a common lipid myopathy. PMID- 11054762 TI - AAEM news and comments PMID- 11054763 TI - Seymour kety, MD. 1915-2000 PMID- 11054764 TI - Vulnerability to addiction: new research opportunities. PMID- 11054765 TI - The D(2) dopamine receptor A(1) allele and opioid dependence: association with heroin use and response to methadone treatment. AB - A total of 95 Caucasian opioid-dependent patients were followed over a one-year period in an outpatient methadone treatment program. The frequency of the TaqI A(1) allele of the D(2) dopamine receptor (DRD2) gene was 19.0% in these patients compared with 4.6% in controls free of past and current alcohol and other drug abuse and free of family history of alcohol and other drug abuse (p = 0.009). Twenty-two of these patients dropped out of the methadone program (Group A), 54 had a successful treatment (Group B), and 19 had a poor treatment (Group C) outcome. The frequency of the A(1) allele was highest in Group C (42.1%), followed by Group A (22.7%) and was lowest in Group B (9.3%). The more than fourfold higher frequency of the A(1) allele in the poor treatment outcome group compared with the successful treatment outcome group was significant (p = 0.00002). Moreover, the average use of heroin (grams/day) during the year prior to study entry was more than twice as great in patients with the A(1)(+) allele (A(1)/A(1) or A(1)/A(2) genotype) than those with the A(1)(-) allele (A(2)/A(2) genotype) (A(1)(+) allele = 0.55 +/- 0. 10, A(1)(-) allele = 0.25 +/- 0.05; p = 0.003). The results indicate that DRD2 variants are predictors of heroin use and subsequent methadone treatment outcome and suggest a pharmacogenetic approach to the treatment of opioid dependence. PMID- 11054766 TI - Confirmation of an excess of the high enzyme activity COMT val allele in heroin addicts in a family-based haplotype relative risk study. AB - A previous case control study by Vandenbergh et al. [1997: Am J Med Genet 74:439 442] showed an association between the high activity catechol O-methyltransferase (COMT) polymorphism and polysubstance abuse in a group of North American subjects. In the current study we confirm these results by genotyping 38 Israeli heroin addicts and both parents using a robust family-based haplotype relative risk (HRR) strategy. There is an excess of the val COMT allele (likelihood ratio = 4.48, P = 0.03) and a trend for an excess of the val/val COMT genotype (likelihood ratio = 4.97, P = 0.08, 2 df) in the heroin addicts compared to the HRR control group. We also genotyped an additional 101 nonrelated heroin addicts and 126 control subjects using a case control design and found no significant difference in COMT val allele frequency (25.4% vs. 29.7%, likelihood ratio = 1.04, P = 0.31). A significant difference is observed in COMT allele frequency among the three principal Israeli ethnic groups (Ashkenazi Jewish, non-Ashkenazi Jewish, and Palestinian Arab) in a large group of control subjects we have so far examined (chi-square = 7.9, P = 0.019, df = 2, n = 1,422 alleles) suggesting that population stratification is responsible for our failure to observe an excess of the COMT val allele when using the case-control design. PMID- 11054767 TI - Detection of single nucleotide polymorphisms of the human mu opioid receptor gene by hybridization or single nucleotide extension on custom oligonucleotide gelpad microchips: potential in studies of addiction. AB - The human mu opioid receptor (MOR) plays a central role in mediating the effects of opioids, both endogenous and exogenous. Epidemiological studies have shown that addiction in general, and especially opiate addiction, has a heritable component. Clinical and laboratory studies suggest that the MOR gene may contribute to the heritable component of vulnerability to develop opiate addiction. Naturally occurring single nucleotide polymorphisms (SNPs) have been identified in the MOR gene by conventional methods. Two coding region SNPs, the A118G and C17T substitutions, occur at high allelic frequencies (10.5% and 6.6%, respectively, in our previous studies). These common SNPs cause amino acid changes in the receptor, and may have implications for differences in individual responses to opioids, as well as decreased or increased vulnerability to opiate addiction. The A118G substitution encodes a variant receptor with binding and signal transduction differences in response to beta-endorphin in cellular assays. Recent innovations in microchip technology offer new potential methods for SNP detection. We report here on the development of two separate approaches using custom oligonucleotide gelpad microarrays for detection of these two common SNPs of the MOR gene in human DNA samples. First, PCR-amplified genomic DNA samples were used to produce target sequences, which were labeled with fluorescent dye and hybridized to custom microchips. Oligonucleotides on these reusable microchips were designed to query nucleotide substitutions at positions 17 and 118 of the MOR gene. Thirty-six human DNA samples were assayed both on these custom microchips and by conventional automated gel sequencing, with highly concordant identification of both heterozygous and homozygous substitutions. A second approach was developed for the C17T SNP utilizing single nucleotide extension on custom microchips. These custom gelpad microchips have potential for the rapid and inexpensive detection of specific SNPs for genetic and genomic studies. PMID- 11054768 TI - Association analysis of polymorphisms in the DRD4 gene and heroin abuse in Chinese subjects. AB - Heroin abuse is a major social and public health problem in many parts of the world, yet relatively little is known about its etiology. Although genes play a role in determining susceptibility, they are expected to be of small effect with considerable heterogeneity. Because the dopamine system is involved in reward, its neurotransmitter receptors are candidates for etiological involvement in addiction. In the present study, we examine two polymorphisms in the dopamine D4 receptor, a VNTR in exon III and a point mutation in the promoter (-512C/T) that affects transcriptional efficiency. We examined a sample of 405 heroin-abusing subjects and 304 controls from Sichuan Province, Southwest China. One hundred twenty-one of these cases and 154 controls were previously used in a study of the DRD4 VNTR [Li et al., 1997], and the remainder are newly ascertained. The two polymorphisms were in weak but detectable linkage disequilibrium (1, 418 chromosomes, P < 0.00001, D' = 0.17). When we compared the heroin-abuse group with controls, we found no significant difference between the patients and controls for either polymorphism in the DRD4 gene or their haplotypes. We were also unable to replicate our earlier association between "long" DRD4 alleles and heroin abuse. However, division of the sample by route of administration (nasal inhalers or injectors) produced a significant difference between inhalers and controls for the DRD4 VNTR (six-fold corrected P = 0. 018 by allele) but not for injectors of heroin. The association we observed between inhalers and the DRD4 polymorphism is difficult to interpret, although it is possible that the association is explained by different levels of novelty seeking between the two subgroups. PMID- 11054769 TI - Haplotypes at the DRD2 locus and severe alcoholism. AB - Association studies of the minor TaqI A allele of the D(2) dopamine receptor (DRD2) gene with alcoholism have produced conflicting findings. Failure to assess alcoholics for severity of their disorder and to screen controls for substance use have been proposed as causes for the discrepant results. In the present study, five diallelic sites spanning the DRD2 gene were determined in combined Caucasian (non-Hispanic) studies of more severe alcoholics (n = 92) and controls screened for substance use (n = 85). The frequency of the minor alleles at the 3' untranslated site (TaqI A) and two intronic sites (TaqI B and intron 6) of the DRD2 gene were each strongly associated with alcoholism. Moreover, the alcoholics compared with the controls at these three sites had a significantly higher frequency of the minor/major allele heterozygote haplotype combination (A1/A2 B1/B2 T/G) than the major allele homozygote haplotype combination (A2/A2 B2/B2 G/G). However, exon 7 and promoter alleles were not associated with alcoholism. In neither the alcoholics nor in the controls were there departures from Hardy Weinberg equilibrium at any of the five sites examined. The most significant diallelic composite genotypic disequilibria were found when comparisons were made between TaqI A and TaqI B, TaqI A and intron 6, and TaqI B and intron 6 sites. Weaker but still significant disequilibria were observed when TaqI A and exon 7, TaqI B and exon 7, intron 6 and exon 7, and promoter and exon 7 sites were compared. However, no significant disequilibria were noted when TaqI A and promoter, TaqI B and promoter, and intron 6 and promoter sites were compared. In sum, the study found significant evidence for association of the minor alleles in the untranslated sites of the DRD2 gene and their haplotypes with the more severe alcoholic phenotype. PMID- 11054770 TI - A genome screen of maximum number of drinks as an alcoholism phenotype. AB - The Collaborative Study on the Genetics of Alcoholism (COGA) is a multicenter research program to detect and map susceptibility genes for alcohol dependence and related phenotypes. The measure M of "maximum number of drinks consumed in a 24-hour period" is closely related to alcoholism diagnosis in this dataset and provides a quantitative measure to grade nonalcoholic individuals. Twin studies have shown log(M) to have a heritability of approximately 50%. Genome screens for this trait were performed in two distinct genotyped samples (wave 1 and wave 2), and in the combined sample. MAPMAKER/SIBS was used to carry out Haseman-Elston based regression analyses. On chromosome 4, an unweighted all-pairs multipoint LOD of 2.2 was obtained between D4S2407 and D4S1628 in wave 1; in wave 2, the region flanked by D4S2404 and D4S2407 gave a LOD of 1.5. In the combined sample, the maximal LOD was 3.5 very close to D4S2407. This evidence for linkage is in the region of the alcohol dehydrogenase gene cluster on chromosome 4. These findings on chromosome 4 are consistent with a prior report from COGA in which strictly defined nonalcoholic subjects in wave 1 were analyzed. The present analysis on log(M) allows more individuals to be included and thus is potentially more powerful. PMID- 11054771 TI - Functional variants at CYP2A6: new genotyping methods, population genetics, and relevance to studies of tobacco dependence. AB - Cytochrome P450CYP2A6 (CYP2A6) is the predominant enzyme responsible for the metabolism of nicotine to cotinine. Two variants have been identified that encode products presumed to have little or no activity. A previous study suggested that carriers of at least one copy of either null variant may be protected against tobacco dependence, while tobacco-dependent carriers smoke fewer cigarettes. However, different laboratories have reported widely disparate CYP2A6 allele frequencies across European populations. These differences prompted us to reexamine the genotyping methods for CYP2A6. We developed an improved genotyping strategy using CYP2A6-specific nested PCR, and differential restriction enzyme digestion to identify variant nucleotides in exon 3. We used sequencing to verify genotype results and to assess the sequence of exon 4, which previous work predicted should correspond to "wild-type" CYP2A6 sequence. In addition, we developed a new nomenclature in which CYP2A6*1 is designated CYP2A6*A1-*B1, CYP2A6*2 is CYP2A6*A2, and CYP2A6*3 is CYP2A6*B2. The frequencies of CYP2A6*A2 and CYP2A6*B2 were then estimated in samples from six populations. Sequencing confirmed CYP2A6*A2 genotypes in all cases. Unexpectedly, sequencing demonstrated exon 4 sequence corresponding to CYP2A7 in samples genotyped as CYP2A6*B2. In the population study, we found consistently low allele frequencies ( or =22 years) age groups. Each completed the Bulimic Investigatory Test and a Stroop task, designed to determine biases in the processing of negative and positive emotional information. RESULTS: Among the older women, more bulimic attitudes were associated with a specific attentional bias toward negatively valenced emotional material. In contrast, the younger women with more severe bulimic attitudes showed greater cognitive avoidance of both positive and negative emotion cues. DISCUSSION: These findings suggest that women with bulimic attitudes differ with age in their processing of emotional information. Potential implications are considered for clinical work with bulimic women of different ages. PMID- 11054783 TI - Attachment patterns in eating disorders: past in the present. AB - OBJECTIVE: There is a wide literature suggesting abnormal mother-daughter and familial attachment patterns in individuals with eating disorders. We surmised that this insecurity would extend to adult attachment relationships. METHODS: The Reciprocal Attachment Questionnaire (RAQ) was administered to all inpatients and outpatients at a tertiary referral eating disorders unit over a given period of time, and to controls. The RAQ operationalizes the key components of reciprocal attachment, and is in close theoretical agreement with the Adult Attachment Interview. RESULTS: Patients scored significantly higher than controls on most scales of the RAQ, most notably on Compulsive Care-Seeking and Compulsive Self Reliance. We did not find any associations between eating disorder diagnoses and particular attachment profiles. CONCLUSIONS: A basic "pull-push" dilemma was demonstrated in the reciprocal attachment relationships of eating-disordered subjects. This dilemma bedevils attempts at therapy and may illuminate the strong feelings elicited by these patients in their therapists. The association of attachment style with particular disorder subgroup diagnoses is complicated. Childhood attachment insecurity may provide a vulnerability whose symptomatic manifestation is colored by later events. PMID- 11054784 TI - Impact of definitions on the description and prediction of bulimia nervosa outcome. AB - OBJECTIVE: The authors sought to compare definitions of eating disorder outcome found in the bulimia nervosa literature and to determine the impact of definitions on the description and prediction of outcome. METHOD: Definitions of outcome were identified from studies involving a follow-up duration of at least 5 years. Definitions were applied to a sample of women (N = 173) assessed more than 10 years following presentation with bulimia nervosa. RESULTS: Across definitions, the percentage of women considered fully recovered ranged from 38% to 47% in the follow-up sample. Associations between eating disorder outcome and other measures of outcome were relatively unaffected by differences in definitions. Conversely, the significance of various prognostic variables differed substantially among definitions. DISCUSSION: Our findings suggest that differences in definitions influence the description and prediction of eating disorder outcome significantly. Consistency in defining recovery is needed in order to explore other areas of outcome such as relapse. PMID- 11054785 TI - Development of an instrument to assess readiness to recover in anorexia nervosa. AB - OBJECTIVE: The degree to which patients with anorexia nervosa demonstrate readiness to recover from their illness has received scant theoretical or empirical attention. Investigating the prevalence and degree of amotivation for recovery in anorexia nervosa, its role in outcome, and the effectiveness of interventions designed to enhance readiness to recover necessitates the existence of a reliable and valid measure of motivational issues relevant to the disorder. The present study aimed to develop and evaluate an instrument for assessing readiness to recover in anorexia nervosa, namely, the Anorexia Nervosa Stages of Change Questionnaire (ANSOCQ), a 23-item self-report questionnaire based on Prochaska and DiClemente's stages of change model. METHOD: Seventy-one inpatients with anorexia nervosa participated in the study. On several occasions during their admission, participants completed the ANSOCQ as well as questionnaires assessing readiness to recover, anorexic symptomatology, general distress, and social desirability. RESULTS: The ANSOCQ demonstrated good internal consistency (.90) and 1-week test-retest reliability (.89). Various aspects of validity were also supported, such as significant relationships with other instruments assessing readiness to recover and the prediction of weight gain during different periods of treatment. DISCUSSION: The results suggest that the ANSOCQ is a psychometrically sound instrument that may prove useful in investigating the role of readiness to recover in anorexia nervosa. PMID- 11054786 TI - Reconceptualization of body image and drive for thinness. AB - OBJECTIVE: This study examined the nature of body image and drive for thinness as multidimensional constructs. Subjects included 111 early adolescent (ages 11-13) girls in Grades 7 and 8 from a private school in Melbourne. METHOD: The participants completed a suite of perceptual, affective/attitudinal, and behavioral measures that included assessment of body mass index, self-concept, body parts, silhouette discrepancy, self-worth, multidimensional body image, body image dissatisfaction, social physique anxiety, eating attitudes and behaviors, and physical activity. RESULTS: The sample reported moderate levels of body image dissatisfaction and a significant association of body image dissatisfaction with drive for thinness. Findings verified the important contribution of the affective/attitudinal components of body image. DISCUSSION: A multidimensional scale devised to test the value of a combined index of self-perceived size, shape, weight, tone, and appearance proved the most effective predictor among the alternative affective/attitudinal scales of body image dissatisfaction. Drive for thinness, as central to this study, was found to be related to, yet distinct from, body image on the basis of behavioral elements such as dieting and activity levels. These findings have ramifications for the design of future research in the body image, drive for thinness, and disordered eating domains. 2000 by John Wiley & Sons, Inc. PMID- 11054787 TI - Body image in the old order Amish: a people separate from "the world". AB - OBJECTIVE: Body image measures were assessed among the Old Order Amish, a Protestant religious community living separate from Western industrialized society. METHOD: One hundred six Old Order Amish men (n = 50) and women (n = 56), aged 14-67 years, were studied by two measures of body image: (1) body dissatisfaction as assessed by the difference between subjects' body mass index (BMI, kg/m(2)) and the BMI that they chose as their ideal and (2) the relative accuracy of perception of body size assessed by comparing subjects' choice of body size on a Figure Rating Scale with the choice of a relative. RESULTS: Young persons and persons of normal weight of both genders showed no body dissatisfaction or inaccuracy in their perception of their body size. Older persons of both genders, on the other hand, manifested body dissatisfaction (actual BMI greater than ideal BMI). Older women also overestimated their body size. Obese persons of both genders manifested body dissatisfaction (actual BMI greater than ideal BMI) and obese men overestimated their body size. DISCUSSION: Young Amish people do not show the body image problems characteristic of young persons in Western industrial society. Their elders and obese persons may have some such problems. 2000 by John Wiley & Sons, Inc. PMID- 11054788 TI - Development of the mizes anorectic cognitions questionnaire-revised: psychometric properties and factor structure in a large sample of eating disorder patients. AB - OBJECTIVE: This project was designed to develop and test the psychometric properties and factor structure of a revision of the Mizes Anorectic Cognitions questionnaire (MAC). The goals of the revision were to improve the reliability and discriminant validity of the Weight and Approval subscale and to equalize the length of the three subscales. Also, the study compared the original MAC and the MAC-R in terms of their psychometric properties. METHOD: Twenty-four new items were developed for potential inclusion in the MAC-R, in addition to the original 33 items of the MAC. These items were administered to 205 eating disorder patients from five eating disorder clinics or programs, including inpatient, outpatient, and residential treatment settings that served diverse patient populations. Additionally, other measures of eating disorder constructs were administered to assess construct validity. RESULTS: Factor analysis of the large pool of items and item reduction resulted in the final 24-item MAC-R, each subscale being eight items in length. Results showed that the MAC-R highly correlated with the MAC and other eating disorder questionnaires. Reliability of the MAC-R was improved over that of the MAC. Two subscales of the MAC-R discriminated among diagnostic groups, whereas the original MAC did not, indicating improved sensitivity of the revised scale. DISCUSSION: The MAC-R appears to be an improvement over the original MAC. It provides useful information on the cognitions of eating-disordered patients and merits further investigation into its psychometric properties. PMID- 11054789 TI - Effect of body image and self-image on women's sexual behaviors. AB - OBJECTIVES: To evaluate the relationship between women's body image, self-image, and sexual behaviors; to determine the effect of body image on personal and sex variables; and to identify factors that contribute to more frequent and greater comfort with sex. METHOD: A magazine survey that included items about body image, self-image, and sexual behaviors was completed by 3,627 women. RESULTS: Women more satisfied with body image reported more sexual activity, orgasm, and initiating sex, greater comfort undressing in front of their partner, having sex with the lights on, trying new sexual behaviors, and pleasing their partner sexually than those dissatisfied. Positive body image was inversely related to self-consciousness and importance of physical attractiveness, and positively related to relationships with others and overall satisfaction. Body image was predictive only of one's comfort undressing in front of partner and having sex with lights on. Overall satisfaction was predictive of frequency of sex, orgasm, and initiating sex, trying new sexual behaviors, and confidence in giving partner sexual pleasure. DISCUSSION: Results indicate that overall self-image and body image are significant predictors of sexual activity. Directions for future research are discussed. PMID- 11054790 TI - A cross-cultural examination of weight-related teasing, body image, and eating disturbance in Swedish and Australian samples. AB - OBJECTIVE: To evaluate, cross-culturally, a model for the prediction of eating disturbance from factors such as body image disturbance, negative verbal feedback regarding appearance (teasing), and body mass index (BMI). METHODS: Three samples of adolescent girls from Sweden (Grade 8: n = 260; mean age = 14.3) and Australia (Grade 7: n = 159; mean age = 12.8 and Grade 8: n = 210; mean age = 13.7) completed two measures of eating restraint and one scale each reflective of bulimic symptomatology, teasing history, and body dissatisfaction. RESULTS: Path analyses revealed that BMI predicted teasing and body dissatisfaction, and body dissatisfaction predicted level of eating restraint. In all three samples, there was evidence of partial mediation by teasing of the connection between BMI and restraint. DISCUSSION: The results partially replicate previous work with U.S. samples. The findings are discussed with regard to the need for further cross cultural work and its relevance for identifying factors for early intervention and prevention programs. PMID- 11054791 TI - Effects of limited access to a fat option on food intake and body composition in female rats. AB - OBJECTIVE: The present investigation sought to determine if limiting access to an optional fatty food would induce binge-type behavior patterns in non-energy deprived female rats. METHOD: Four groups of rats had continuous access to a commercial rodent diet throughout the 8-week study. In addition: (1) the control group had no access to vegetable shortening; (2) the high limitation group had access to shortening for 2 hr for 3 days each week; (3) the low limitation group had access to shortening for 2 hr every day; and (4) the no limitation group had continuous access to shortening. RESULTS: As access to the shortening decreased, intake during the 2-hr access period increased. Total energy intake and body weight did not differ among groups. Body fat was greatest in the rats that ate the most cumulative shortening. DISCUSSION: These results indicate that, even under non-energy-deprived conditions, limiting access to a preferred fatty food can induce binge-type behavior in female rats. PMID- 11054792 TI - Anger and bulimic psychopathology among nonclinical women. AB - OBJECTIVE: Although there are well-established links between bulimic psychopathology and some affective states, the role of anger is not clearly understood. This is likely to be a product of the diverse nature of anger. The present study examines the association of different components of anger with bulimic attitudes and behaviors among a nonclinical group of women. METHODS: Eighty-three nonclinical women completed standardized measures of anger and bulimic attitudes/behaviors. RESULTS: Bulimic attitudes and behaviors were correlated specifically with state anger and anger suppression, rather than with trait anger. The pattern of results suggests that binging and vomiting behaviors may serve different functions with regard to anger. CONCLUSIONS: Bulimic attitudes and behaviors appear to reduce immediate anger states, particularly when the individual has a strong tendency to avoid expressing that emotion. Potential therapeutic implications are considered. PMID- 11054793 TI - Prevalence of thiamin deficiency in anorexia nervosa. AB - OBJECTIVE: Deficiency of thiamin (vitamin B1) causes a range of neuropsychiatric symptoms that resemble those reported in patients with anorexia nervosa (AN) but the prevalence of thiamin deficiency in AN has not been reliably established. This study was designed to investigate the prevalence of thiamin deficiency in AN. METHOD: Thirty-seven patients attending a specialist eating disorders unit and meeting all or some of the DSM-IV criteria for AN were compared with 50 blood donor controls. All subjects underwent measurement of erythrocyte transketolase activation following the addition of thiamin pyrophosphate, the standard biochemical test for thiamin deficiency. Deficiency was defined as a result more than 2 SD above the mean of the control population. RESULTS: Fourteen patients (38%) had results in the deficient range; 7 (19%) met the most stringent published criterion for deficiency. Deficiency was not related to duration of eating restraint, frequency of vomiting, or alcohol consumption. DISCUSSION: Thiamin deficiency may account for some of the neuropsychiatric symptoms of AN and routine screening or supplementation may be indicated. PMID- 11054794 TI - Comparison of the yale-brown-cornell eating disorders scale in recovered eating disorder patients, restrained dieters, and nondieting controls. AB - OBJECTIVE: Yale-Brown-Cornell Eating Disorder Scale (YBC-EDS) scores were assessed in recovered eating disorder patients, restrained dieters, and unrestrained nondieters. METHOD: YBC-EDS interviews were conducted with 53 recovered eating disorder patients who had no symptoms within at least 6 months, 29 restrained dieters, and 36 unrestrained controls. RESULTS: Unrestrained control subjects had no typical eating-disordered preoccupations or rituals. The majority (62%) of restrained dieters did have current eating-disordered preoccupations but only 5 had current eating-disordered rituals. Most recovered eating disorder subjects had no current eating-disordered preoccupations (66%) and 76% had no current eating-disordered rituals. Unrestrained eating controls had significantly lower Preoccupation, Total, and Motivation to Change scores on the YBC-EDS than the other groups and significantly lower Ritual scores than the recovered eating disorder group. There were no significant differences between the restrained dieters and the recovered eating disorder group. DISCUSSION: Recovered eating disorder patients who no longer meet any of the DSM-IV criteria for an eating disorder are similar in severity of eating concern to normal weight restrained eating dieters. Both of these groups have more eating and weight concerns as compared with the unrestrained eating, nondieting controls. The YBC EDS effectively distinguishes the healthy eating controls from restrained eating dieters and recovered eating disorder patients. PMID- 11054795 TI - Ice-cream consumption, tendency toward overeating, and personality. AB - OBJECTIVE: The exploration of the mechanisms underlying the tendency toward overeating by investigating the Dutch Eating Behavior Questionnaire (DEBQ)/Revised Eating Disorders Inventory (EDI-R) disinhibition, in sequence to the milkshake-ice cream study (van Strien, Cleven, and Schippers, in press). METHOD: In hierarchical multiple regression analyses, the relative predictive power for ice-cream consumption was assessed, that is, emotional versus external versus bulimic eating using scales of the DEBQ and the EDI-R. In nonplanned stepwise multiple regression analyses, the association was assessed between these three types of eating behaviors and non-eating-related EDI-R scales. RESULTS: Emotional eating was the most important variable for ice-cream consumption. External eating was borderline significant and bulimic eating nonsignificant when emotional and external eating had been partialled out. Emotional eating was best predicted by the EDI-R scales Asceticism, Interoceptive Awareness, and Social Insecurity. DISCUSSION: Results are consistent with psychosomatic theory, which focuses on emotional eating as the result of confusion and apprehension in recognizing and accurately responding to emotional and visceral states related to hunger and satiety. PMID- 11054796 TI - Assessment of eating disorders in bariatric surgery candidates: self-report questionnaire versus interview. AB - OBJECTIVE: To compare the Eating Disorder Examination (EDE), an investigator based interview for the assessment of the specific psychopathology of eating disorders, with the EDE-Q, a self-report questionnaire based directly on it. METHOD: Ninety-eight morbidly obese gastric bypass surgery candidates were administered both instruments. RESULTS: The four subscale scores (Restraint, Eating Concern, Weight Concern, and Shape Concern) generated by the EDE and EDE-Q were significantly correlated, although the questionnaire scores were significantly higher. Eating Concern and Shape Concern exhibited the lowest levels of agreement. Frequency of binges (objective bulimic episodes) as rated by the EDE and EDE-Q was significantly correlated and was not significantly different. However, variability in ratings contributed to only modest agreement with respect to classification of patients as binge eaters. DISCUSSION: Overall, there were lower levels of agreement between the EDE and EDE-Q than have been previously found in other samples. PMID- 11054797 TI - Enteric feeding in severe adolescent anorexia nervosa: a report of four cases. AB - OBJECTIVE: Refeeding patients with anorexia nervosa can be one of the more challenging aspects of their treatment, and particularly if all food and fluids are adamantly and persistently refused. METHOD: If the decision is made to augment or replace oral feeds, the most common intervention is nasogastric feeding. RESULTS: Although this is often successful, a subset of patients manage to sabotage feeding via this route. Other means of delivering nutrition such as intravenous feeds are often impractical for long-term use. Another alternative in such life-threatening situations is the use of enteric feeds via gastrostomy or jejunostomy. This paper presents the successful use of such enteric feeding in four cases of severe adolescent anorexia nervosa. DISCUSSION: The psychological, legal, and ethical issues involved are discussed, concluding that gastrostomy and jejunostomy are valid lifesaving methods to feed highly resistant anorectic patients. PMID- 11054798 TI - "Spiritual starvation?": a case series concerning christianity and eating disorders. AB - METHOD: We describe the cases of four patients with eating disorders in whom complex interactions occurred among religious faith, pathogenesis of the eating disorder, and clinical management. RESULTS: In some of the cases, religious beliefs seemed to provide a containment of maladaptive behaviors, partly through prayer and through a sense of belonging to the religious community. In other cases, it proved difficult to separate the concept of a punitive God from the illness process. DISCUSSION: The cases are discussed with reference to a limited empirical literature. Similarities are noted between some religious institutions and eating disorder treatment regimes. This paper explores management issues, including the use of pastoral counseling and the ethics of addressing religious beliefs in therapy. We note the benefits of a rapprochement between psychiatry and religion. PMID- 11054799 TI - Erratum PMID- 11054800 TI - Platelet-derived growth factor and basic fibroblast growth factor regulate cell proliferation and the expression of notch-1 receptor in a new oligodendrocyte cell line. AB - We generated a new cell line, N38, by conditionally immortalizing mouse oligodendrocytes (OLs) at early stages of maturation. The morphology and marker expression pattern suggest N38 cells are similar to immature OLs. N38 cells were sensitive to changes in serum concentrations, and forcing the cells to differentiate in low serum at 39 degrees C significantly decreased the survival of the cells. Importantly, addition of PDGFaa, bFGF or astrocyte-conditioned medium had protective effects on the cells, by increasing cell proliferation but not cell differentiation. This effect was receptor-mediated. Exposure of N38 cells to differentiating signals such as retinoic acid did not cause further differentiation of the cells. The N38 cell line expresses the vertebrate homolog of the Drosophila notch-1 receptor, a molecule that appears to regulate OL differentiation. Notch-1 receptor was homogeneously distributed in the somas of N38 cells. Incubation of N38 cells with either PDGFaa or bFGF, however, induced a polarized distribution of the receptor in the majority of the cells as well as an upregulation of receptor protein levels. The upregulation of molecules, such the notch-1 receptor, in pathways that control differentiation might be an important mechanism for keeping OL precursors in an undifferentiated state during their exit of the germinal layer and migration in the developing central nervous system. This OL cell line might constitute a suitable model for studies of regulatory mechanisms at this stage of OL differentiation. PMID- 11054801 TI - NT-3 weakly stimulates proliferation of adult rat O1(-)O4(+) oligodendrocyte lineage cells and increases oligodendrocyte myelination in vitro. AB - The transplantation of fibroblasts, genetically modified to secrete neurotrophin 3 (NT-3) and/or brain-derived neurotrophic factor (BDNF), into spinal cord injured rats increases the production of new oligodendrocytes and myelination (McTigue et al. [1998] J. Neurosci. 18:5354-5365). This experiment did not fully resolve whether the effect was exerted on oligodendrocyte precursors or on oligodendrocytes, or whether there was stimulation of both proliferation and differentiation of the oligodendrocyte lineage cells. To clarify the effects of NT-3 and BDNF, adult rat spinal cord was dissociated to produce cultures in which both oligodendrocyte precursors (O1(-)O4(+)) and oligodendrocytes (O1(+)) were present. Thymidine labeling of cells was determined in the presence and absence of added NT-3 and/or BDNF. In addition, the effect of these neurotrophins on myelination was determined by treating purified adult O1(+) oligodendrocyte/embryonic dorsal root ganglion (DRG) neuron cocultures with neurotrophins, only during the myelination period. O1(+) oligodendrocyte proliferation was not stimulated by NT-3 or BDNF; however, the proliferation of O1(-)O4(+) cells was increased in NT-3-treated cultures to a labeling index (LI: 24 hr) of 15-20%. This effect was observed at 5 but not at 10 days in vitro. In comparison, basic fibroblast growth factor (bFGF) induced the proliferation of both O1(+) oligodendrocytes (LI approximately 60%) and O1(-)O4(+) cells (LI approximately 75%). The amount of myelin formed in purified O1(+) oligodendrocyte/DRG neuron cocultures was significantly increased in NT-3-treated cultures compared to untreated cultures. These results indicate that NT-3 is weakly but transiently mitogenic for adult-derived oligodendrocyte precursors and support the suggestion that NT-3 promotes the maturation of O1(+) oligodendrocytes into myelin-forming cells. PMID- 11054802 TI - Distinct roles for PI3K in proliferation and survival of oligodendrocyte progenitor cells. AB - Phosphoinositol 3-kinase (PI3K) is a downstream effector for multiple ligand activated receptors and modulates cell responses through activation of its target protein kinase B (Akt). We examined the roles of PI3K-Akt signaling in a primary glial (oligodendrocyte) progenitor cell culture system that is ligand-dependent for cell proliferation, survival, and prevention of differentiation. We demonstrate that PI3K and Akt (Ser-473 phosphorylation) are activated in response to platelet-derived growth factor but not basic fibroblast growth factor-2 (FGF2) and that distinct forms of PI3K are activated in early progenitors and later maturation pro-oligodendroblasts as identified by their sensitivity to wortmannin. By establishing conditions to examine effects on cell proliferation and survival independently, we demonstrate that PI3K is necessary for a full mitogenic response and that PI3K is also necessary for early progenitor survival. Our results therefore demonstrate that PI3K-Akt signaling independently regulates proliferation and survival, that the form of PI3K is distinct in early progenitors and pro-oligodendroblasts, and that FGF2 does not activate this pathway in either primary glial cell population. PMID- 11054803 TI - Expression and subcellular localization of two isoforms of the survival motor neuron protein in different cell types. AB - The survival motor neuron (SMN) gene is deleted or mutated in over 98% of spinal muscular atrophy patients who show specific motoneuron loss. By performing transfection experiments with rat smn cDNA, we show that two isoforms of SMN with Mr of 32 kDa and 35 kDa are produced by the same cDNA. In cultured motoneurons, both forms colocalize in coiled bodies and not in GEMS bodies as shown for HeLa cells. Subcellular fractionation of cells acutely dissociated from rat embryonic ventral spinal cord shows that the two SMN isoforms have a different subcellular localization, namely, that the 32 kDa isoform is enriched in the cytosol, whereas the 35 kDa isoform is segregating in the microsomal fraction. We show that the 35 kDa isoform of SMN is part of an insoluble complex but is absent from the cytoplasmic membranes and from the mitochondria. Immunostaining studies show that neither SMN isoform colocalizes with Bcl-2, the mitochondrial antiapoptotic protein suggested to bind to SMN in HeLa cells. Our results show that the isoforms of SMN protein have different subcellular localization and may therefore play independent biological roles. Moreover, the absence of colocalization of SMN with Bcl-2 in motoneurons suggests that some of the interactors of SMN may vary depending on the cell type, and this underscores the importance of identifying motoneuron-specific SMN interactors. PMID- 11054804 TI - Brain-derived neurotrophic factor increases activity of NR2B-containing N-methyl D-aspartate receptors in excised patches from hippocampal neurons. AB - Growth factors, including members of the neurotrophin gene family, play a central role in the regulation of neuronal survival and differentiation during development. In addition to these relatively long-term actions of neurotrophins, recent studies have shown that these factors also rapidly modulate synaptic transmission. Brain-derived neurotrophic factor (BDNF), in particular, regulates both pre- and postsynaptic aspects of hippocampal synaptic transmission. The postsynaptic effects include an increase in glutamate responsiveness, mediated by the N-methyl-D-aspartate (NMDA) glutamate receptor subtype. It is not clear, however, where BDNF-trkB signal transduction is initiated, because trkB receptors are located in both pre- and postsynaptic membranes. In the present study, we used excised membrane patches from cultured hippocampal neurons to determine whether BDNF directly modulates postsynaptic NMDA receptor activity. The results indicate that acute exposure to BDNF increases NMDA single channel open probability via postsynaptic trkB receptors and that this effect is dependent on the presence of the NR2B subunit of the NMDA receptor. PMID- 11054805 TI - Expression of gangliosides in neuronal development of P19 embryonal carcinoma stem cells. AB - Gangliosides are constituents of the cell membrane and are known to have important functions in neuronal differentiation. We employed an embryonal carcinoma stem cell line P19 as an in vitro model to investigate the expression of gangliosides during neuronal development. After treatment with retinoic acid, these cells differentiate synchronously into neuron-like cells by a series of well-defined events of development. We examined several aspects of ganglioside metabolism, including the changes of ganglioside pattern, the activities and gene expression of several enzymes at different stages of differentiation, and the distribution of gangliosides in differentiating neurons. Undifferentiated P19 cells express mainly GM3 and GD3. After P19 cells were committed to differentiation, the synthesis of complex gangliosides was elevated more than 20 fold, coinciding with the stage of neurite outgrowth. During the maturation of differentiated cells, the expression of c-series gangliosides was downregulated concomitantly with upregulation of the expression of a- and b-series gangliosides. We also examined the distribution of gangliosides in differentiating neurons by confocal and transmission electron microscopy after cholera toxin B subunit and sialidase treatment. Confocal microscopic studies showed that gangliosides were distributed on the growth cones and exhibited a punctate localization on neurites and soma. Electron microscopic studies indicated that they also are enriched on the plasma membranes of neurites and the filopodia as well as on the lamellipodia of growth cones during the early stage of neurite outgrowth. Our data demonstrate that the expression of gangliosides in P19 cells during RA-induced neuronal differentiation resembles that of the in vivo development of the vertebrate brain, and hence validates it as an in vitro model for investigating the function of gangliosides in neuronal development. PMID- 11054806 TI - Effect of 1,25-dihydroxyvitamin D(3) on cultured mesencephalic dopaminergic neurons to the combined toxicity caused by L-buthionine sulfoximine and 1-methyl 4-phenylpyridine. AB - A decrease in intracellular glutathione content may be related to the primary event in Parkinson's disease, so increasing the glutathione level may have a therapeutic benefit. The biologically active form of vitamin D, 1,25 dihydroxyvitamin D(3) [1, 25-(OH)(2)D(3)] has been recently reported to enhance the intracellular glutathione concentration in the central nervous system. Exposing rat cultured mesencephalic neurons for 24 hr to a mixture of L buthionine sulfoximine (BSO) and 1-methyl-4-phenylpyridium ions (MPP(+)) resulted in a relatively selective damage to dopaminergic neurons. This damage has been accompanied by a reduction of intracellular glutathione levels. Low doses, i.e., 1-100 nM, of 1,25-(OH)(2)D(3) protect cultured dopaminergic neurons against this toxicity, although higher concentrations of this active form of vitamin D have been found to enhance the toxic effect. Generation of reactive oxygen species (ROS) by this toxicity has been attenuated in cultures being pretreated with low concentrations of 1,25-(OH)(2)D(3). Because the hormone increases the intracellular glutathione content in cultures, determining how this hormone suppresses ROS generation may involve the enhancement of the antioxidative system. These data suggest that low doses of 1,25-(OH)(2)D(3) are able to protect mesencephalic dopaminergic neurons against BSO/MPP(+)-induced toxicity that causes a depletion in glutathione content. PMID- 11054807 TI - Differentiated regulation of allo-antigen presentation by different types of murine microglial cell lines. AB - We established granulocyte-macrophage colony-stimulating factor (GM-CSF) dependent murine microglial clones and investigated the immune properties of four individual clones. All four clones expressed MHC class I and CD54 (ICAM-1) at similar levels. The 5-2, Ra2, and 6-3 clones expressed CD80 (B7-1), CD86 (B7-2), and MHC class II at low, medium, and high levels, respectively. Only the 6-3 clone expressed CD40. Generally, the levels of co-stimulation and CD 40 signals had a profound effect on the response to antigens. The 5-2, Ra2, and 6-3 clones, however, stimulated allogenic T-cell proliferation to the same extent or less compared to spleen cells. Although the 6-1 clone expressed co-stimulatory and MHC molecules at levels similar to Ra2, it suppressed allogenic T-cell proliferation, unlike Ra2. Thus, allo-antigen presentation by microglial clones was not correlated with the expression of CD40 and co-stimulatory molecules. When microglial clones were fixed with paraformaldehyde, they enhanced IL-2-dependent T-cell proliferation according to the level of their expression of co-stimulatory molecules. Furthermore, conditioned medium from the 6-1 clone inhibited the T cell response to allo-antigen. This indicates that some factor(s) derived from a microglial subtype may play an important role in the regulation of T-cell proliferation in addition to the molecules involved in antigen presentation. Moreover, these results also suggest that there may be specialized subtypes of microglia that regulate the immune response in the CNS. PMID- 11054808 TI - Induction of hepatocyte growth factor (HGF) in rat microglial cells by prostaglandin E(2). AB - Hepatocyte growth factor (HGF) is a multifunctional protein that exerts trophic effects on neural cells. HGF is expressed in normal brains and increased after brain injury. Recent studies suggest that neurons and astrocytes are the main producers of HGF in the brain. Here we report that microglia also produce HGF both in vitro and in vivo. Treatment of cultured microglia with prostaglandin E(2) (PGE(2)), one of the major inflammatory mediators in the brain, induced significant production of HGF, and this induction was suppressed by pretreatment with the adenylate cyclase inhibitor SQ22536, suggesting that the induction of HGF by PGE(2) in microglia proceeds via a cAMP-mediated pathway. We further investigated whether microglia also produce HGF in vivo under the pathological condition of cerebral ischemia. We found that HGF expression was increased after permanent occlusion of the middle cerebral artery (MCA), and double immunohistochemical staining revealed that the most of HGF-positive cells were microglia. PGE(2) level was increased 8 hr after start of MCA occlusion, and this enhancement is in parallel with the increase in HGF expression, suggesting that PGE(2) not only may induce HGF production in microglia in vitro but may also be an inducer in vivo. PMID- 11054809 TI - Farnesol modulates membrane currents in human retinal glial cells. AB - Farnesol, a C(15) natural isoprenoid, exerts complex modulating effects on the membrane permeability of human retinal glial (Muller) cells. Several glial cationic currents were examined. At low micromolar concentrations, farnesol reduced the amplitudes of all fast and depolarization-activated membrane currents expressed by Muller cells, that is, currents through 1) transient low-voltage activated (LVA; IC(50) = 2.2 microM), 2) sustained high-voltage-activated Ca(2+) channels (HVA; IC(50) = 1.2 microM), 3) fast Na(+) channels (IC(50) = 9.0 microM), and 4) transient (A-type) K(+) channels (IC(50) = 4.7 microM). Furthermore, farnesol shifted the activation of LVA and HVA currents to more depolarized potentials by 21.3 +/- 7.4 mV and 8.3 +/- 4.5 mV, respectively. On the other hand, neither inwardly rectifying nor iberiotoxin-sensitive calcium activated K(+) currents were affected by farnesol. Therefore, farnesol is assumed to be a biologically active substance that regulates ion channel activity in the glial cell membrane. Depressing rapid changes of the membrane potential and supporting a stable hyperpolarized status of the glial cells may enhance the efficiency of crucial glial functions such as extracellular K(+) clearance and neurotransmitter uptake. PMID- 11054810 TI - Influence of an extracellular acidosis on excitatory synaptic transmission and long-term potentiation in the CA1 region of rat hippocampal slices. AB - The effects of extracellular acidification on the synaptic function and neuronal excitability were investigated on the hippocampal CA1 neurons. A decrease of extracellular pH from 7.4 to 6.7 did not alter either the resting membrane potential or the neuronal membrane input resistance. Extracellularly recorded field excitatory postsynaptic potentials (fEPSPs) and population spikes (PSs) were significantly reduced by acidosis. Additionally, the amplitude of presynaptic fiber volley was also reduced. The sensitivity of postsynaptic neurons to N-methyl-D-aspartate, but not to alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid, was depressed by acidosis. Lowering of extracellular pH did not significantly affect the magnitude of paired-pulse facilitation (PPF) of synaptic transmission. Acidosis also reversibly limited the sustained repetitive firing (RF) of Na(+)-dependent action potentials elicited by injection of depolarizing current pulses into the pyramidal cells. The limitation of RF by extracellular acidification was accompanied by the reduction of the maximal rate of rise (;V(max)) of the action potentials and the amplitude of afterhyperpolarization. Neither the Na (+)/H (+) antiporter blocker 5-(N -ethyl N -isopropyl)-amiloride nor the selective adenosine A (1) receptor antagonist 1,3 dipropyl -8-cyclopentylxanthine, however, affected the acidosis -induced synaptic depression. It was also found that acidosis did not affect either the induction r maintenance of long -term potentiation (LTP) at Schaffer collateral -CA 1 synapses. These results suggest that the extracellular acidosis -induced synaptic depression is likely to result from an inhibition of presynaptic Na (+) conductance, thereby decreasing the amplitude of action potentials in individual afferent fibers or the number of afferent fiber activation to stimuli and then indirectly affecting the signaling processes contributing to trigger neurotransmitter release. PMID- 11054811 TI - Differential regulation of primary protein kinase C substrate (MARCKS, MLP, GAP 43, RC3) mRNAs in the hippocampus during kainic acid-induced seizures and synaptic reorganization. AB - In the mature hippocampus, kainic acid seizures lead to excitotoxic cell death and synaptic reorganization in which granule cell axons (mossy fibers) form ectopic synapses on granule cell dendrites. In the present study, we examined the expression of four major, developmentally regulated protein kinase C (PKC) substrates (MARCKS, MLP, GAP-43, RC3), which have different subcellular and regional localizations in the hippocampus at several time points (6 hr, 12 hr, 18 hr, 24 hr, 48 hr, 5 days, or 15 days) following kainic acid seizures using in situ hybridization. Consistent with previous reports, following kainate seizures, GAP-43 mRNA expression exhibited a delayed and protracted elevation in the granule cell layer, which peaked at 24 hr, whereas expression in fields CA1 and CA3 remained relatively unchanged. Conversely, RC3 mRNA expression exhibited a delayed reduction in the granule cell layer that was maximal at 18 hr, as well as a reduction CA1 at 48 hr, whereas CA3 levels did not change. MARCKS mRNA expression in the granule cell layer and CA1 remained stable following kainate, although an elevation was observed in subfield CA3c at 12 hr. Similarly, MLP mRNA expression did not change in the granule cell layer or CA1 following kainate but exhibited a protracted elevation in subfields CA3b,c beginning at 6 hr post kainate. Collectively these data demonstrate that different PKC substrate mRNAs exhibit unique expression profiles and regulation in the different cell fields of the mature hippocampus following kainic acid seizures and during subsequent synaptic reorganization. The expression profiles following kainate seizures bear resemblance to those observed during postnatal hippocampal development, which may indicate the recruitment of common regulatory mechanisms. PMID- 11054812 TI - Possible role for the FosB/JunD AP-1 transcription factor complex in glutamate mediated excitotoxicity in cultured cerebellar granule cells. AB - The potent excitatory and neurotoxic actions of glutamate are known to influence the expression of a variety of genes, including those encoding the AP-1 transcription factor, which comprises proteins belonging to the Fos and Jun families. However, the precise role of Fos- and Jun-like transcription factors in these events remains elusive. Here we demonstrate, using primary cultures of mouse brain cerebellar granule cells as an in vitro model system, a possible involvement of the FosB/JunD heterodimer in excitotoxicity. Granule cells were grown for either 2 or 7 days in vitro (DIV) before exposure to varying concentrations (1-3000 microM) of the excitotoxin glutamate. In 7-DIV cells, glutamate induced a concentration-dependent neuronal death, whereas, in 2-DIV cells, no glutamate-induced neuronal damage was seen. We were particularly interested in comparing the protein composition of the AP-1 transcription factor complex in cells exposed to excitotoxic and to nontoxic conditions. AP-1 DNA binding activity was demonstrated by gel shift analysis in nuclear extracts derived from 7-DIV cells following exposure to either a nontoxic (10 microM) or an excitotoxic (250 microM) dose of glutamate and was similarly observed in extracts of 2-DIV cells exposed to the same levels of glutamate. Gel supershift analysis using antibodies against the different Fos and Jun family members allowed differentiation between AP-1 DNA binding in nuclear extracts as a function of both 1) viability status and 2) the stage of development. Of major significance was the finding that FosB could be detected as a component of AP-1 in 7-DIV cells only under excitotoxic conditions, whereas c-Fos, Fra-2, and JunD proteins were detectable under both excitotoxic and nontoxic conditions in cells of this age. In 2-DIV cells (in which glutamate is nontoxic), AP-1 comprised combinations of only Fra-1, Fra-2, c-Jun, and JunD. Because Fos family members are unable to form homodimers, this finding raises the possibility that the FosB/JunD heterodimer may have special significance in the mechanism of excitotoxic neuronal death. PMID- 11054813 TI - Evidence for oxidative damage in a murine leukemia virus-induced neurodegeneration. AB - Vacuolation in cellular organelles within the central nervous system is a common manifestation of oxidative injury. We found that the spongiform vacuolation observed in PVC-211 murine leukemia virus (PVC-MuLV) neurodegeneration was associated with oxidative damage as detected by immunoreactivity for 3 nitrotyrosine and protein carbonyl groups. This oxidative injury was present in brain before or concomitant with the appearance of activated microglia, vacuolation, and gliosis that characterize PVC-MuLV neuropathology. Treatment of infected F344 rat pups with the antioxidant vitamin E transiently protected and prolonged the latency of PVC-MuLV neurodegeneration. Taken together, these findings implicate oxidative damage and lipid peroxidation in the pathogenesis of PVC-MuLV neurodegeneration. This animal model may be useful for studies of mechanisms and potential therapies for progressive neurodegeneration following a well-defined insult. PMID- 11054814 TI - Laminin inhibition of beta-amyloid protein (Abeta) fibrillogenesis and identification of an Abeta binding site localized to the globular domain repeats on the laminin a chain. AB - beta-Amyloid protein (Abeta) is a major component of neuritic plaques and cerebrovascular amyloid deposits in the brains of patients with Alzheimer's disease (AD). Inhibitors of Abeta fibrillogenesis are currently sought as potential future therapeutics for AD and related disorders. In the present study, the basement membrane protein laminin was found to bind Abeta 1-40 with a single dissociation constant, K(d) = 2.7 x 10(-9) M, and serve as a potent inhibitor of Abeta fibril formation. 25 microM of Abeta 1-40 was incubated at 37 degrees C for 1 week in the presence of 100 nM of laminin or other basement membrane components, including perlecan, type IV collagen, and fibronectin to determine their effects on Abeta fibril formation as evaluated by thioflavin T fluorometry. Of all the basement membrane components tested, laminin demonstrated the greatest inhibitory effect on Abeta-amyloid fibril formation, causing a ninefold inhibition at 1 and 3 days and a 21-fold inhibition at 1 week. The inhibitory effects of laminin on Abeta fibrillogenesis occurred in a dose-dependent manner and were still effective at lower concentrations. The inhibitory effects of laminin on Abeta 1-40 fibril formation was confirmed by negative stain electron microscopy, whereby laminin caused an almost complete inhibition of Abeta fibril formation and assembly by 3 days, resulting in the appearance of primarily amorphous nonfibrillar material. Laminin also caused partial disassembly of preformed Abeta-amyloid fibrils following 4 days of coincubation. Laminin was not effective as an inhibitor of islet amyloid polypeptide fibril formation, suggesting that laminin's amyloid inhibitory effects were Abeta-specific. To identify a potential Abeta-binding site(s) on laminin, laminin was first digested with V8, trypsin, or elastase. An Abeta-binding elastase digestion product of approximately 120-130 kDa was found. In addition, a approximately 55 kDa fragment derived from V8 and elastase-digested laminin interacted with biotinylated Abeta 1-40. Amino acid sequencing of the approximately 55 kDa fragment identified a conformationally dependent Abeta-binding site within laminin localized to the globular repeats on the laminin A chain. These studies demonstrate that laminin not only binds Abeta with relatively high affinity but is a potent inhibitor of Abeta-amyloid fibril formation. In addition, further identification of an Abeta binding domain within the globular repeats on the laminin A chain may lead to the design of new therapeutics for the inhibition of Abeta fibrillogenesis. PMID- 11054815 TI - Phosphorylation of human tau protein by microtubule-associated kinases: GSK3beta and cdk5 are key participants. AB - Microtubules (MTs), primarily composed of alpha and beta tubulin polymers, must often work in concert with microtubule-associated proteins (MAPs) in order to modulate their functional demands. In a mature brain neuron, one of the key MAPs that resides primarily in the axonal compartment is the tau protein. Tau, in the adult human brain, is a set of six protein isoforms, whose binding affinity to MTs can be modulated by phosphorylation. In addition to the role that phosphorylation of tau plays in the "normal" physiology of neurons, hyperphosphorylated tau is the primary component of the fibrillary pathology in Alzheimer's disease (AD). Although many protein kinases are known to phosphorylate tau in vitro, the in vivo players contributing to the hyperphosphorylation of tau remain elusive. The experiments in this study attempt to define which protein kinases and protein phosphatases reside in the associated network of microtubules, thereby being strategically positioned to influence the phosphorylation of tau. Microtubule fractions are utilized to determine which of the microtubule-associated kinases most readily impacts the phosphorylation of tau at "AD-like" sites. Results from this study indicate that PKA, CK1, GSK3beta, and cdk5 associate with microtubules. Among the MT-associated kinases, GSK3beta and cdk5 most readily contribute to the ATP-induced "AD-like" phosphorylation of tau. PMID- 11054816 TI - Progression of cell cycle monitored by dielectric spectroscopy and flow cytometric analysis of DNA content. AB - A dielectric method has already been developed for the real-time monitoring of cell cycle progression in synchronized cell culture (Asami et al., 1999). This method, in combination with DNA content analysis by fluorescence-activated cell sorting (FACS), was applied to the synchronized cell culture of a CDC28-13th mutant (Saccharomyces cerevisiae). In synchronous cell growth, relative permittivity epsilon (or dielectric constant) for the culture broth showed cyclic changes at low frequencies below 0.5 MHz, being correlated to phases in the cell cycle that were simultaneously determined by FACS. The epsilon increased in the period from S phase to G(2) phase and decreased between M and G(1) phases. Peaks in these cyclic changes of epsilon indicated the time when daughter cells segregated from mother cells. PMID- 11054817 TI - Constitutive activation of the Saccharomyces cerevisiae transcriptional regulator Ste12p by mutations at the amino-terminus. AB - The transcriptional activator Ste12p is required for the expression of genes induced by mating pheromone in the yeast Saccharomyces cerevisiae. We identified mutations in the amino-terminal DNA-binding domain of Ste12p that lead to constitutively high-level transcription of pheromone-induced genes. The behaviour of these mutant proteins is consistent with an enhanced DNA-binding ability. Cells carrying these hyperactive proteins retain their sensitivity to pheromone treatment, and their phenotype is largely dependent on the presence of at least one of the MAP kinases (Fus3p or Kss1p) and the scaffold protein Ste5p. Deletion of either FUS3 or KSS1 leads to a marked increase in Ste12p activity, consistent with a negative regulatory role for Fus3p, similar to that described for Kss1p. The properties of the constitutive mutants support the idea that the pheromone response pathway plays a role in basal as well as pheromone-induced transcription. PMID- 11054818 TI - Organization of specific genomic regions of Zygosaccharomyces rouxii and Pichia sorbitophila: comparison with Saccharomyces cerevisiae. AB - The genomes of Zygosaccharomyces rouxii and Pichia sorbitophila were partially explored. The genome of Z. rouxii CBS 732 consists of seven chromosomes with an approximate size of 1.0-2.75 Mb, 12.8 Mb in total. Five of the chromosomes were labelled with specific probes. Three Z. rouxii genomic DNA fragments were sequenced; all 10 ORFs found were without introns and they have homologues in S. cerevisiae. Gene order comparison revealed that the organization is partially conserved in both species. The genome of P. sorbitophila CBS 7064 consists of seven chromosomes with an approximate size of 1.0-2.9 Mb, 13.9 Mb in total. Three of the chromosomes were labelled with specific probes. The sequencing of a 5.2 kb genomic DNA fragment revealed three ORFs, but no conservation of their organization was found, although all of them have their respective homologues in S. cerevisiae. According to our results, the presence of two overlapping ORFs in S. cerevisiae (YJL107c-YJL108c) could be interpreted as the result of a frameshift mutation. PMID- 11054819 TI - Functional reconstitution of an active recombinant human chymase from Pichia pastoris cell lysate. AB - We have previously reported efficient production of mature human chymase (h chymase) using an original system of expression in Pichia pastoris (Nakakubo et al., 2000), whereby recombinant h-chymase (rh-chymase) was secreted as a mature form with the correct N-terminal amino acid sequence and was easily purified. In the course of investigation of secretory rh-chymase, we also found large amounts of chymase to be present in insoluble form in the transformant cell. Although the cellular rh-chymase had no proteolytic activity, its chymotryptic activity was restored in a reconstitution process utilizing guanidine and glutathione. As with secretory rh-chymase, efficient purification was possible by heparin affinity chromatography. The purified cellular rh-chymase showed the same mobility as secretory rh-chymase in sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) before and after deglycosylation. N-terminal amino acid sequence analysis revealed that the signal peptide had been correctly removed. K(m) value (5.93 mM), as well as pH profile and inhibition profile toward protease inhibitors of reconstituted cellular rh-chymase, indicated that the rh-chymase enzymatically closely resembles native h-chymase. Furthermore, it showed a greatly restricted proteolytic activity towards Ang I, and formed Ang II without the further cleavage which is a feature of h-chymase. It was thus found that the insoluble rh-chymase stored in the cells could be solubilized and reconstituted to give the same structure as h-chymase, not only in terms of enzyme active site but also of substrate recognition site. PMID- 11054820 TI - Characterization of the L41 gene in Cryptococcus neoformans: its application as a selectable transformation marker for cycloheximide resistance. AB - A transformation system using resistance to the antibiotic cycloheximide as a dominant selectable marker was developed for the pathogenic yeast Cryptococcus neoformans. A 3.5 kb DNA fragment containing a gene encoding the ribosomal protein L41 was cloned from a wild-type strain of C. neoformans which is sensitive to cycloheximide. The open reading frame of the L41 gene contains five introns and encodes a protein of 107 amino acids, which is similar to those reported for other yeasts. The cycloheximide resistance gene to be used as a marker was constructed by replacing a DNA segment of the wild-type L41 gene, which contained the amino acid proline at its 56th position with a homologous DNA segment from a mutant strain resistant to cycloheximide that contained leucine in that position. Cycloheximide resistant transformants were obtained by electroporation on YEPD plates, supplemented with 10-20 microg/ml cycloheximide, at a maximum efficiency of 300 transformants/microg plasmid DNA. While with other genes, most transformants of serotype D in C. neoformans maintain the transforming DNA as episomes, the cycloheximide-resistant transformants were all the result of ectopic genomic integration events. PMID- 11054821 TI - Analysis of 114 kb of DNA sequence from fission yeast chromosome 2 immediately centromere-distal to his5. AB - One hundred and fourteen kilobase pairs (kb) of contiguous genomic sequence have been determined immediately distal to the his5 genetic marker located about 0.9 Mb from the centromere on the long arm of Schizosaccharomyces pombe chromosome 2. The sequence is contained in overlapping cosmid clones c16H5, c12D12, c24C6 and c19G7, of which 20 kb are identical to previously reported sequence from clone c21H7. The remaining 93 781 bp of sequence contains 10 known genes (cdc14, cdm1, cps1, gpa1, msh2, pck2, rip1, rps30-2, sad1 and ubl1), 32 open reading frames (ORFs) capable of coding for proteins of at least 100 amino acid residues in length, one 5S rRNA gene, one tRNA(Pro) gene, one lone Tf1-type long terminal repeat (LTR) and one lone Tf2-type LTR. There is a density of one protein-coding gene per 2.2 kb and 22 of the 42 ORFs (52%) incorporate one or more introns. Twenty-one of the novel ORFs show sequence similarities which suggest functions of their products, including a cyclin C, a MADS box transcription factor, mad2 like protein, telomere binding protein, topoisomerase II-associated protein, ATP dependent DEAH box RNA helicase, G10 protein, ubiquitin-activating e1-like enzyme, nucleoporin, prolyl-tRNA synthetase, peptidylprolyl isomerase, delta-1 pyrroline-5-carboxylate dehydrogenase, protein transport protein, coatomer epsilon, TCP-1 chaperonin, beta-subunit of 6-phosphofructokinase, aminodeoxychorismate lyase, a phosphate transport protein and a thioredoxin. PMID- 11054822 TI - Molecular cloning and characterization of the Candida albicans UBI3 gene coding for a ubiquitin-hybrid protein. AB - Using a polyubiquitin cDNA as a probe, we have isolated a clone (pPR3, a pEMBLYe23 derivative plasmid) containing the Candida albicans UBI3 gene coding for a fusion protein. This protein is formed by one ubiquitin subunit fused, at its C-terminus, to an unrelated peptide which is similar to the ribosomal protein encoded by the 3' tail of the Saccharomyces cerevisiae UBI3 gene. Southern blot analysis of chromosomal DNA probed with the 3' non-ubiquitin tail of UBI3 indicated that only one homologous gene is present in the C. albicans genome. Heterelogous expression of pPR3 in a S. cerevisiae ubi3 mutant strain complements the mutant phenotype (slow growth) conferred by the ubi3 defect; this provides direct evidence indicating that the clone contains the C. albicans UBI3 gene Northern blot analysis showed that UBI3 gene is expressed in yeast and germ-tube cells of C. albicans, although the UBI3 mRNA levels in starved yeast cells are below the detection limit; UBI3 mRNA drops to undetectable levels on shifting the temperature of growing yeast cells from 28 degrees C to 42 degrees C. When Northern blot analysis was performed using a specific probe for the polyubiquitin (UBI4) gene, no drop in the mRNA levels was detected following thermal upshift or in starved cells. These results indicate that stress conditions (starvation or thermal upshift) negatively regulate UBI3 expression (transcriptional arrest and/or enhanced mRNA decay), and suggest that UBI4 gene provides ubiquitin during the stress response. In addition, we failed to obtain C. albicans UBI3 null mutant cells by sequential disruption of both alleles using the hisG::URA3::hisG ('ura-blaster') cassette, suggesting that null mutants cells may be unable to grow on selective media after transformation. PMID- 11054823 TI - Mitochondria-targeted green fluorescent proteins: convenient tools for the study of organelle biogenesis in Saccharomyces cerevisiae. AB - We describe the construction and characterization of a novel set of plasmids for expression of mitochondria-targeted green fluorescent protein (GFP) in Saccharomyces cerevisiae. The vectors include constructs with strong regulatable and constitutive promoters, four different auxotrophic markers for yeast transformation, and a green (S65T) and a blue-shifted (P4-3) variant of GFP. Mitochondria are brightly fluorescent in living yeast cells grown on different carbon sources and at different temperatures, with virtually no background staining. Specific staining of mitochondria is also shown for a respiratory deficient mutant with abnormal mitochondrial morphology. The plasmids facilitate convenient analysis of mutants defective in mitochondrial morphology or inheritance and, at the same time, are suitable vectors for easy construction of different kinds of GFP fusion proteins to study various aspects of organelle biogenesis in living yeast cells. PMID- 11054824 TI - Disruption and functional analysis of six ORFs on chromosome XII of saccharomyces cerevisiae: YLR124w, YLR125w, YLR126c, YLR127c, YLR128w and YLR129w. AB - In the context of the EUROFAN programme, we report the deletion and functional analysis of six open reading frames (ORFs) on the right arm of chromosome XII of Saccharomyces cerevisiae. Using a PCR-based gene replacement strategy, we have systematically deleted individual ORFs and subjected the heterozygous diploids and haploid knockout strains to basic genetic and phenotypic characterization. Two ORFs, YLR127c and YLR129w, are essential for viability, whereas no growth phenotype could be detected following deletion of YLR124w, YLR125w, YLR126c or YLR128w. For each of the individual ORFs, a kanMX4 replacement cassette and the corresponding cognate wild-type gene were cloned into appropriate plasmids. PMID- 11054825 TI - Disruption and functional analysis of six ORFs on chromosome IV: YDL053c, YDL072c, YDL073w, YDL076c, YDL077c and YDL080c. AB - Six open reading frames (ORFs) from chromosome IV, YDL053c, YDL072c, YDL073w, YDL076c, YDL077c and YDL080c, were disrupted using the long flanking homology technique (LFH) to replace each target locus with the KanMX4 selection marker. We have also constructed plasmids containing replacement cassettes (pYORC) and the cognate clones (pYCG) for each ORF. Disruption of five of the ORFs-YDL053c, YDL072c, YDL073w, YDL076c and YDL080c (THI3)-resulted in no distinctive phenotype with respect to temperature or nutritional requirements. However, disruption of YDL077c (also known as VAM6) exhibited an slow growth phenotype in minimal media and also in rich media containing glycerol as a carbon source. The homozygous disruptant diploid corresponding to this gene also failed to sporulate. PMID- 11054826 TI - Current awareness on yeast. AB - In order to keep subscribers up-to-date with the latest developments in their field, this current awareness service is provided by John Wiley & Sons and contains newly-published material on yeasts. Each bibliography is divided into 10 sections. 1 Books, Reviews & Symposia; 2 General; 3 Biochemistry; 4 Biotechnology; 5 Cell Biology; 6 Gene Expression; 7 Genetics; 8 Physiology; 9 Medical Mycology; 10 Recombinant DNA Technology. Within each section, articles are listed in alphabetical order with respect to author. If, in the preceding period, no publications are located relevant to any one of these headings, that section will be omitted. (4 weeks journals - search completed 23rd Aug. 2000) PMID- 11054827 TI - Different behavior toward racemization in basic media from chiral analogs of clofibric acid, the active metabolite of the antilipidemic drug clofibrate. AB - Some chiral analogs of clofibric acid, the active metabolite of the antilipidemic drug clofibrate, show different configurational stability in basic conditions. Also, extensive racemization occurs when the corresponding optically active acid chlorides are treated with 3 alpha-tropanol, whereas no racemization takes place with 3 alpha-tropanol as hydrochloride salt and with 3 beta-tropanol and 1-methyl 4-hydroxy-piperidine as either the free base or hydrochloride salt. For these aminoalcohols, experimental evidence supports the hypothesis that a ketene intermediate is involved in the racemization process. Formation of intramolecular hydrogen bond is evoked to explain the different ability of aminoalcohols to induce ketene formation and consequent racemization. PMID- 11054828 TI - Resolution, configurational assignment, and enantiopharmacology of 2-amino-3-[3 hydroxy-5-(2-methyl-2H- tetrazol-5-yl)isoxazol-4-yl]propionic acid, a potent GluR3- and GluR4-preferring AMPA receptor agonist. AB - We have previously shown that (RS)-2-amino-3-[3-hydroxy-5-(2-methyl-2H-tetrazol-5 yl)isoxazol -4-yl] propionic acid (2-Me-Tet-AMPA) is a selective agonist at (RS) 2-amino-3-(3-hydroxy-5-methylisoxazol-4-yl)propionic acid (AMPA) receptors, markedly more potent than AMPA itself, whereas the isomeric compound 1-Me-Tet AMPA is essentially inactive. We here report the enantiopharmacology of 2-Me-Tet AMPA in radioligand binding and cortical wedge electrophysiological assay systems, and using cloned AMPA (GluR1-4) and kainic acid (KA) (GluR5, 6, and KA2) receptor subtypes expressed in Xenopus oocytes. 2-Me-Tet-AMPA was resolved using preparative chiral HPLC. Zwitterion (-)-2-Me-Tet-AMPA was assigned the (R) configuration based on an X-ray crystallographic analysis supported by the elution order of (-)- and (+)-2-Me-Tet-AMPA using four different chiral HPLC columns and by circular dichroism spectra. None of the compounds tested showed detectable affinity for N-methyl-D-aspartic acid (NMDA) receptor sites, and (R)-2 Me-Tet-AMPA was essentially inactive in all of the test systems used. Whereas (S) 2-Me-Tet-AMPA showed low affinity (IC(50) = 11 microM) in the [(3)H]KA binding assay, it was significantly more potent (IC(50) = 0.009 microM) than AMPA (IC(50) = 0.039 microM) in the [(3)H]AMPA binding assay, and in agreement with these findings, (S)-2-Me-Tet-AMPA (EC(50) = 0.11 microM) was markedly more potent than AMPA (EC(50) = 3.5 microM) in the electrophysiological cortical wedge model. In contrast to AMPA, which showed comparable potencies (EC(50) = 1.3-3.5 microM) at receptors formed by the AMPA receptor subunits (GluR1-4) in Xenopus oocytes, more potent effects and a substantially higher degree of subunit selectivity were observed for (S)-2-Me-Tet-AMPA: GluR1o (EC(50) = 0.16 microM), GluR1o/GluR2i (EC(50) = 0.12 microM), GluR3o (EC(50) = 0.014 microM) and GluR4o (EC(50) = 0.009 microM). At the KA-preferring receptors GluR5 and GluR6/KA2, (S)-2-Me-Tet-AMPA showed much weaker agonist effects (EC(50) = 8.7 and 15.3 microM, respectively). It is concluded that (S)-2-Me-Tet-AMPA is a subunit-selective and highly potent AMPA receptor agonist and a potentially useful tool for studies of physiological AMPA receptor subtypes. PMID- 11054829 TI - Synthesis and characteristics of the human serum albumin-triazine chiral stationary phase. AB - Human serum albumin (HSA) was successfully bonded to silica with s-triazine as activator. The coupling reaction by this method was rapid and effective. The triazine-activated silica is relatively stable and can be installed for at least 1 month without obvious loss of reactivity when stored below 30 degrees C, pH below 7. It was observed that the amount of bound HSA reached 120 mg/g silica calculated from the UV absorbance difference of the HSA solution. d, l-tryptophan was selected as the probe solute to characterize the properties of HSA bonded s triazine chiral stationary phase, and separation factor of 9.4 was obtained for d,l-tryptophan. Furthermore, the amount of effective HSA on silica was measured by high-performance frontal analysis, and only 16.8 mg/g silica was responsible for the resolution of d,l-tryptophan. These results indicate that the amount of both the bound and effective HSA on silica with triazine as activator was much higher than those by the Schiff base coupling method. Different kinds of enantiomers were resolved successfully on the aminopropylsilica-bonded HSA s triazine chiral stationary phase. PMID- 11054830 TI - Stereochemical study of tolperisone, a muscle relaxant agent, by circular dichroism and ultraviolet spectroscopy. AB - The stereochemistry of tolperisone, a chiral aryl-alkyl basic ketone was investigated by means of circular dichroism (CD) and ultraviolet (UV) spectroscopy. The unusually high optical activity of tolperisone hydrochloride in the n-->pi* region is interpreted by the presence of a chiral conformer in solution. For stereochemical reasons, the C = O group and the aromatic moiety lack coplanarity by forming an inherently dissymetric chromophore, of M helicity. Similar helicity prevails in the crystal phase, according to the solid-state CD spectrum of (-)-tolperisone HCl salt. The chirality rule proposed by Snatzke for nonplanar benzoyl chromophores predicts the absolute configuration of (-) tolperisone hydrochloride to be R, in agreement with other alpha-methyl-beta amino-ketones. PMID- 11054831 TI - HPLC enantiomeric resolution of novel aromatase inhibitors on cellulose- and amylose-based chiral stationary phases under reversed phase mode. AB - The chiral resolution of seven aromatase inhibitors (four triazole derivatives (Ia, Ib, Ic, and Id) and three tetrazole derivatives (IIa, IIb, and IIc)) was achieved on Chiralcel OJ-R [cellulose tris (4-methyl benzoate)], Chiralcel OD-RH [cellulose tris (3,5-dimethylphenyl carbamate)], and Chiralpak AD-RH [amylose tris (3,5-dimethylphenyl carbamate)] chiral stationary phases. The mobile phases used were A: 2-PrOH-MeCN (90:10, v/v); B: 2-PrOH-MeCN (50:50, v/v); C: MeCN-H(2)O (50:50, v/v); D: MeCN-H(2)O (80:20, v/v); and E: MeCN-H(2)O (95:05, v/v). The flow rate was 0.5 mL/min for all the mobile phases. The resolution capability of these chiral stationary phases were in the order Chiralpak AD-RH > Chiralcel OD RH > Chiralcel OJ-R. The values of alpha and Rs of the resolved enantiomers of the aromatase inhibitors varied from 1.02-5.63 and 1. 12-6.72, respectively. PMID- 11054832 TI - Chiral overcrowded alkenes; asymmetric synthesis of (3S,3'S)-(M, M)-(E)-(+) 1,1',2,2',3,3',4,4'-octahydro-3,3',7,7'-tetramethyl-4, 4'-biphenanthrylidenes. AB - An asymmetric synthesis route towards (3S,3'S)-(M,M)-(E)-(+)-1,1',2, 2',3,3',4,4' octahydro-3,3',7,7'-tetramethyl-4,4'-biphenanthrylidene was developed using the Evans procedure as a key step. The absolute configurations of the title compound and of its parent ketone were determined by CD spectroscopy and could be correlated with the stereochemical results of the asymmetric alkylation. Furthermore, a comparison was made with the known (3R,3'R)-(P,P)-(E)-(-) 1,1',2,2', 3,3',4,4'-octahydro-3,3',7,7'-dimethyl-4,4'-biphenanthrylidene. Finally, the X-ray crystallographic analysis of (3S,3'S)-(M, M)-(E)-(+) 1,1',2,2',3,3',4,4'-octahydro-3,3',7,7'-tetramethyl-4, 4'-biphenanthrylidene is presented. PMID- 11054833 TI - Comparative molecular field analysis of quinine derivatives used as chiral selectors in liquid chromatography: 3D QSAR for the purposes of molecular design of chiral stationary phases. AB - A comparative molecular field analysis (CoMFA) was carried out on a set of aligned quinine-based stationary phase molecules used in enantioselective chromatography. The best QSAR derived has a cross-validated (predictive) r(2)(cv) = 0.671 and a normal r(2) = 0.998. For CoMFAs using both steric and electrostatic fields as descriptors, the steric field descriptors explained more than 91% of the variance while the electrostatic descriptors explained less than 9% of the variance. It is concluded that the long-range electrostatic potential surrounding the positively charged CSPs are not enantiodiscriminating, while the van der Waals and local electrostatic surface features of these CSPs are highly discriminating. Quantum mechanical calculations back up this claim by showing a relatively symmetric electrostatic iso-contour surface. From the QSAR derived here, a region around the carbamate moiety was located where placement of steric bulk is predicted to enhance chiral discrimination. A set of possible synthetic target molecules is presented. PMID- 11054834 TI - Separation and absolute configuration of the enantiomers of a degradation product of the new inhalation anesthetic sevoflurane. AB - In a rebreathing anesthesia circuit, the inhaled anesthetic sevoflurane degrades into at least two products, termed "compound A" and "compound B." The enantiomer separation of the chiral compound B (1,1,1,3,3-pentafluoro-2-(fluoromethoxy)-3 methoxypropane ) by capillary gas chromatography (cGC) using heptakis (2,3-di-O acetyl-6-O-tert-butyldimethylsilyl)-beta-cyclodextrin as chiral selector was studied. With this cyclodextrin derivative diluted in the polysiloxane PS 86, an unprecedented high separation factor alpha of 4.1 (at 30 degrees C) was found. Consequently, the enantiomers of compound B were isolated by preparative GC and their specific rotations were measured. In addition, their absolute configurations were determined by X-ray crystallography. To collect the X-ray data, single crystals of both enantiomers were grown in situ on the diffractometer. The levorotatory enantiomer B(-) has the R-configuration while the dextrorotatory enantiomer B(+) has the S-configuration. The elution order of the compound B enantiomers on heptakis (2,3-di-O-acetyl-6-O-tert butyldimethylsilyl)-beta-cyclodextrin is R before S. PMID- 11054835 TI - Ethnobotany and its role in drug development. AB - The botanical collections of early explorers and the later ethnobotany have played important roles in the development of new drugs for many centuries. In the middle of the last century interest in this approach had declined dramatically, but has risen again during its last decade, and new foci have developed. The systematic evaluation of indigenous pharmacopoeias in order to contribute to improved health care in marginalized regions has been placed on the agenda of international and national organizations and of NGOs. In this paper the results of various projects on Mexican Indian ethnobotany and some of the subsequent pharmacological and phytochemical studies are summarized. Medicinal plants are an important element of indigenous medical systems in Mexico. This study uses the medicinal plants in four indigenous groups of Mexican Indians-Maya, Nahua, Zapotec and Mixe-as an example. The relative importance of a medicinal plant within a culture is documented using a quantitative method and the data are compared intra- and interculturally. While the species used by the indigenous groups vary, the data indicate that there exist well-defined criteria specific for each culture, which lead to the selection of a plant as a medicine. For example, a large number of species are used for gastrointestinal illnesses by two or more of the indigenous groups. At least in this case, the multiple transfers of species and their uses within -Mexico seems to be an important reason for the widespread use of a species. Some of the data we gathered in order to evaluate the indigenous claims are also discussed, focusing on the transcription factor NF kappaB as a molecular target. This led to the identification of sesquiterpene lactones such as parthenolide as potent and relatively specific inhibitors of this transcription factor. PMID- 11054836 TI - Antioxidant and hepatoprotective effects of Acathopanax senticosus. AB - Acathopanax senticosus (Rupr. et Maxim.) Harms. is a popular folk medicine used as a nutrient for hepatitis and cancer in Taiwan. In this study, the antioxidant activity of the crude extract and the hepatoprotective activities on CCl(4)- or acetaminophen-induced toxicity in the rat liver were evaluated. Our results suggest that A. senticosus exerts some antioxidant effects. On a CCl(4)- or acetaminophen-intoxicated -model, the levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were increased by CCl(4) or acetaminophen administration and reduced by treatment with the plant extract. Histological changes around the hepatic central vein were also recovered by treatments. However, treatments with larger doses of the crude extract of A. senticosus enhanced liver damage. This result suggests that even if A. senticosus had hepatoprotective activity in small doses, treatment with larger doses would possibly induce some cell toxicity. PMID- 11054837 TI - In vitro and in vivo activity of eugenol on human herpesvirus. AB - Eugenol (4-allyl-1-hydroxy-2-methoxybenzene) was tested for antiviral activity against HSV-1 and HSV-2 viruses. In vitro, it was found that the replication of these viruses was inhibited in the presence of this compound. Inhibitory concentration 50% values for the anti-HSV effects of eugenol were 25.6 microg/mL and 16.2 microg/mL for HSV-1 and HSV-2 respectively, 250 microg/mL being the maximum dose at which cytotoxicity was tested. Eugenol was virucidal and showed no cytotoxicity at the concentrations tested. Eugenol-acyclovir combinations synergistically inhibited herpesvirus replication in vitro. Topical application of eugenol delayed the development of herpesvirus induced keratitis in the mouse model. PMID- 11054838 TI - Inhibitory effects of an aqueous extract of Apocynum venetum leaves and its constituents on Cu(2+)-induced oxidative modification of low density lipoprotein. AB - An aqueous extract of Apocynum venetum leaves and its constituents inhibited thiobarbituric acid reactive substances (TBARS) and conjugated-diene formation in the Cu(2+)-induced oxidation of low density lipoprotein (LDL) in vitro. The TBARS formation was most strongly inhibited by chlorogenic acid with an IC(50) value of 1.9 microM, but other constituents were in a range of 2.3-23.3 microM. On the other hand, the lag time in the conjugated-diene formation was dose-dependently prolonged by addition of the aqueous extract. Catechin prolonged the lag time more than 300 min and other constituents such as chlorogenic acid, epicatechin, epigallocatechin, hyperoside and isoquercitrin led to no conjugated-diene formation within 700 min under the experimental conditions. PMID- 11054839 TI - Aconiti tuber (Bushi) improves microcirculatory disturbances induced by endotoxin in rats. AB - The present study investigated the effects of processed Aconiti tuber (TJ-3022, Tsumura-shuchi-bushi), a traditional herbal medicine (Kampo), on microcirculatory disturbances induced by endotoxin in the rat mesentery. The changes in arteriolar diameter, the velocity of red blood cells in arterioles and the venular microcirculation after endotoxin injection (6 mg/kg, iv) were observed using videomicroscopy. The mean arterial pressure and heart rate were monitored continuously during the experiments. TJ-3022 prevented significantly the decrease of mean arterial pressure, controlled the reduced velocity and apparently also inhibited leukocyte extravasation across venules. However, there were no significant changes in the arteriolar diameter and the heart rate between the groups treated with and without TJ-3022. The results indicate that processed Aconiti tuber (TJ-3022) has protective effects in improving microcirculatory disturbances induced by endotoxin in rat mesentery. PMID- 11054840 TI - Inhibitory effects of sudanese medicinal plant extracts on hepatitis C virus (HCV) protease. AB - One hundred fifty-two methanol and water extracts of different parts of 71 plants commonly used in Sudanese traditional medicine were screened for their inhibitory effects on hepatitis C virus (HCV) protease (PR) using in vitro assay methods. Thirty-four extracts showed significant inhibitory activity (>/=60% inhibition at 100 microg/mL). Of these, eight extracts, methanol extracts of Acacia nilotica, Boswellia carterii, Embelia schimperi, Quercus infectoria, Trachyspermum ammi and water extracts of Piper cubeba, Q. infectoria and Syzygium aromaticum, were the most active (>/=90% inhibition at 100 microg/mL). From the E. schimperi extract, two benzoquinones, embelin (I) and 5-O-methylembelin (II), were isolated and found as potent HCV-PR inhibitors with IC(50) values of 21 and 46 microM, respectively. Inhibitory activities of derivatives of I against HCV-PR as well as their effects on other serine proteases were also investigated. PMID- 11054841 TI - Fetoprotectivity of the flavanolignan compound siliphos against ethanol-induced toxicity. AB - Of the three flavanolignans that are found in silymarin (Silybum marianum [L.] Gaertn.), silybin is thought to be the primary therapeutic constituent. To test the capacity of silybin to protect the rat fetus from toxic effects of maternally ingested EtOH we did the following: Adult female rats were assigned to one of four groups; EtOH, EtOH/silybin, pair-fed control, and chow fed control. Silybin was orally administered as Siliphos(R), which is one part silybin to two parts phosphatidylcholine. All groups except the chow-fed control were maintained on a liquid diet throughout pregnancy. On day 21 of pregnancy the rats were killed and the fetuses removed. Gamma glutamyl transpeptidase (GGTP) activity and glutathione (GSH) levels were determined for liver and brain tissue for both the fetuses and the dams. Maternal and fetal GGTP activity in the EtOH rats was significantly higher than that of pair-fed controls, whereas the GGTP activity observed in the Siliphos(R)/EtOH rats was not elevated. Fetal mortality rates in the EtOH rats significantly exceeded those of all three other groups. PMID- 11054842 TI - Antihepatotoxic activity of Rosmarinus tomentosus in a model of acute hepatic damage induced by thioacetamide. AB - R. tomentosus is a vegetal species closely related to the culinary rosemary (R. officinalis), a plant reported to contain antihepatotoxic agents. A dried ethanol extract of the aerial parts of Rosmarinus tomentosus (Lamiaceae) and its major fraction separated by column chromatography (fraction F19) were evaluated for antihepatotoxic activity in rats with acute liver damage induced by a single oral dose of thioacetamide. Silymarin was used as a reference antihepatotoxic substance. Pre-treatment with R. tomentosus ethanol extract, fraction F19 or silymarin significantly reduced the impact of thioacetamide toxicity on plasma protein and urea levels as well as on plasma aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase and gamma-glutamyl transpeptidase activities compared with thioacetamide-treated animals (group T). Pre-treatment with R. tomentosus ethanol extract significantly reduced the impact of thioacetamide damage on alkaline phosphatase and gamma-glutamyl transpeptidase activities compared with group T. Silymarin administration significantly reduced alkaline phosphatase and gamma-glutamyl transpeptidase activities compared with group T. Fraction F19 administration reduced only alkaline phosphatase activity compared with group T. According to these data, R. tomentosus extract shows promising antihepatotoxic activity, suggesting the need to isolate the chemical principles responsible for this activity and to study this activity in a model of thioacetamide-induced cirrhosis. PMID- 11054843 TI - Alterations in the periodontium after ovariectomy in rats: the effects of a Japanese herbal medicine, Chujo-to. AB - Ovariectomy-induced changes on the periodontium (gingiva, alveolar bone, and periodontal ligament) in rats and the preventive effects of a Japanese herbal medicine, Chujo-to, were studied for a period of 49 days. The rats were divided into five groups: sham-operated (sham), ovariectomized (OVX), OVX given Chujo-to, OVX given 17beta-oestradiol, and OVX given the vehicle for 17beta-oestradiol, respectively. After the test period, the bone mineral content (BMC) of the mandibular condyle in OVX rats was similar to those in both sham rats and the OVX rats treated with either Chujo-to or 17beta-oestradiol. However, the scanning electron microscopic (SEM) analyses revealed that the periodontal ligament of the OVX rats and the OVX rats treated with Chujo-to became more coarse than that of the sham rats or the rats treated with 17beta-oestradiol. The surface of the alveolar bone in the OVX rats appeared to contain numerous small granules, which were not present in the sham rats and the rats treated with either Chujo-to or 17beta-oestradiol. These results suggest that ovariectomy caused alterations in the peridontium, but Chujo-to had a preventive effect on the surface architecture of the alveolar bones. PMID- 11054844 TI - Preliminary screening of some traditional Zulu medicinal plants for antineoplastic activities versus the HepG2 cell line. AB - Aqueous and methanol extracts of nine traditional Zulu medicinal plants, Cissus quandrangularis L., Cyphostemma flaviflorum (Sprague) Descoings, Cyphostemma lanigerum (Harv.) Descoings ex Wild & Drum, Cyphostemma natalitium (Szyszyl.) J. v. d. Merwe, Cyphostemma sp., Rhoicissus digitata (L. F.) Gilg & Brandt, Rhoicissus rhomboidea (E. Mey. Ex harv.) Planch, Rhoicissus tomentosa (Lam.) Wild & Drum, R. tridentata (L. F.) Wild & Drum and Rhoicissus tridentata (L. F.) Wild & Drum subsp. cuneifolia (Eckl. & Zeyh.) N. R. Urton, all belonging to the Vitaceae family, were evaluated to determine their therapeutic potentials as antineoplastic agents. The antiproliferative activity in vitro against HepG2 cells was determined. Twenty-two of the twenty-seven crude plant extracts showed activities ranging from 25% to 97% inhibition of proliferation when compared with the control which showed no inhibitory activity. Higher degrees of growth inhibition were found in aqueous root extracts in comparison with the methanol extracts of the same plant parts. The results show potential antineoplastic activity, indicating some scientific validation for traditional usage. PMID- 11054845 TI - Flavonol glycosides from Aristeguietia discolor reduce morphine withdrawal in vitro. AB - The effects of extracts, partially purified fractions and four flavonol glycosides 1-4 from Aristeguietia discolor were investigated on the naloxone precipitated withdrawal contraction of the acute morphine dependent guinea-pig ileum in vitro. After a 4 min in vitro exposure to morphine a strong contraction of guinea-pig isolated ileum was observed after the addition of naloxone. Both MeOH extract (50, 100 and 200 mg/mL), the partially purified fractions I, L, M and N (50, 100 and 200 mg/mL) and flavonol glycosides 1-4 (1 x 10(-4) 5 x 10(-5) 1 x 10(-5) M), injected 10 min before morphine, were capable of blocking the naloxone-induced contraction after exposure to morphine in a concentration dependent fashion. The results of the present paper suggest that flavonol glycosides from Aristeguietia discolor may play an important role in the control of morphine withdrawal. PMID- 11054846 TI - Evaluation of antitussive potential of Jussiaea suffruticosa linn. extract in albino mice. AB - The antitussive activity of a methanol extract of Jussiaea suffruticosa Linn. (MEJS) (family Onagraceae) leaves has been evaluated for its potential on a cough induced by sulphur dioxide (SO(2)) gas model in mice. The extract (MEJS) showed significant antitussive activity in a dose dependent manner. The antitussive potential of MEJS was comparable to that of codeine phosphate (10 mg/kg), a standard drug. The extract (MEJS) at a dose level of 200 and 400 mg/kg, p.o. showed appreciable inhibition on the cough reflex by 48.52% and 59.8% respectively during 120 min of the experiment. PMID- 11054847 TI - Antiulcer effectiveness of Maytenus aquifolium spray dried extract. AB - The antiulcer activity of Maytenus aquifolium spray dried extract was studied in rats. Ulcers were induced by means of three experimental models: acidified ethanol, indomethacin and acute stress. The extract was found to have significant antiulcer activity against all the models studied. These results show that preparation of the extract by means of the spray dried technique does not alter the biological activity of Maytenus aquifolium. PMID- 11054848 TI - Screening for antioxidant properties of Salvia reflexa hornem. AB - The antioxidant properties of the wild growing sage species, Salvia reflexa Hornem., were investigated. The presence of superoxide (O(2)(.-)) and hydroxyl ((.)OH) radicals, malonyldialdehyde (MDA), reduced glutathione (GSH) and total flavonoids were observed in the above-ground parts of plant, as well as activities of the antioxidant enzymes superoxide-dismutase (SOD) and peroxidase (P-ase). The potential antioxidant activity of the methanol:water extract has been assessed based on scavenging activity of stable DPPH free radicals. By the means of TLC and LC/MS a screening for secondary plant products was performed. Significant quantities of O(2)(.-), (.)OH and MDA were observed. Thus, this species exhibited high SOD and P-ase activities as well as a content of total flavonoids. The dominant naturally occurring compound was rosmarinic acid. The results obtained suggested a high antioxidant activity of Salvia reflexa Hornem. PMID- 11054849 TI - Myorelaxant and antispasmodic effects of the essential oil of Alpinia speciosa on rat ileum. AB - The effects of the essential oil of Alpinia speciosa Schum (EOAS) on rat ileum were studied. EOAS (0.1-600 microg/mL) reversibly relaxed ileal basal tonus. Submaximal contractions induced by 60 mM KCl or acetylcholine were concentration dependently inhibited by EOAS with similar IC(50) values ( approximately 44 and 48 microg/mL, respectively). These results show that EOAS possesses both relaxant and antispasmodic actions in the ileum. PMID- 11054850 TI - Cytotoxic activity of Agave intermixta L. (Agavaceae) and Cissus sicyoides L. (Vitaceae). AB - The cytostatic activities of Agave intermixta L. (Agavaceae) and Cissus sicyoides L. (Vitaceae) have been determined. In the antimitotic assay, Agave intermixta L. showed complete inhibition of cell division at 24 h of treatment. Both species showed a moderate cytostatic activity against HEp-2 cells, Cissus sicyoides L. being the most active species. PMID- 11054851 TI - Clastogenic effect of ginger rhizome in mice. AB - The present study focuses on the clastogenic effect of ginger rhizome. Crude aqueous extracts of ginger were gavaged at doses of 0. 5, 1, 2, 5, 10 g/kg body weight and ginger oil (0.625, 1.250 and 2. 50 ml/kg body weight) was administered by intraperitoneal injection to male mice. Chromosome damage was studied in a preparation made from bone marrow cells following colchicine injection to all mice and examination of the cells after pretreatment in hypotonic solution, fixation, air drying and staining in Giemsa solution. Attention is drawn to the weakness of the clastogenic activity expressed by the ginger extract. In comparison ginger oil gave a higher frequency of chromosomal aberrations. It is suggested therefore, that the extract may contain substance(s) that suppress clastogenesis in the bone marrow cells of mice. PMID- 11054852 TI - Antioxidant activity of Artemisia douglasiana besser extract and dehydroleucodine. AB - The charge (relative concentration) of antioxidants in aqueous extract and dehydroleucodine (DhL) from Artemisia douglasiana Besser was determined employing a procedure based on the quenching of luminol-enhanced chemiluminescence. Total reactive antioxidant potential (TRAP) and total antioxidant reactivity (TAR) values were determined. The data reported in the present work indicate that the extract of Artemisia douglasiana Besser and DhL showed antioxidant capacity. This activity is more pronounced in the extract, suggesting the presence of several antioxidants in Artemisia douglasiana. PMID- 11054853 TI - In vitro antimicrobial activity of extracts and isolated constituents of Geum rivale. AB - The antimicrobial activity of extracts of Geum rivale (Rosaceae) and that of some isolated constituents, on bacteria and fungi, was evaluated. The activity was concentrated in the triterpenes fraction and, for gram+ and gram- bacteria, also in the flavonoids fraction. PMID- 11054854 TI - Inhibition of chemically induced liver carcinogenesis in Wistar rats by garlic (Allium sativum). AB - The effects of garlic on diethylnitrosoamine (DEN)-induced hepatocarcinogenesis were examined in male Wistar rats using the medium term bioassay system of Ito based on the two-step model of hepatocarcinogenesis. Carcinogenic potential was scored by comparing the number and area/cm(2) of induced glutathione S transferase placental form (GST-P) positive foci in the liver with those of the corresponding group (control) of rats given diethylnitrosoamine alone. Treatment with garlic (therapeutic dose 20 mg/kg body wt/day) reduced significantly the number (50% reduction, p < 0.003) and area (48% reduction, p < 0.0007) of GST-P positive foci compared with the control group of animals receiving distilled water. Histopathological examination of rat livers using H & E staining indicated that there was no significant difference between the control group and the garlic treated group in the two pathological parameters namely granularity and vacuolation of the cytoplasm. Our results provide strong supportive evidence for the anticarcinogenic activity of garlic. PMID- 11054855 TI - Effect of Withania somnifera glycowithanolides on iron-induced hepatotoxicity in rats. AB - Glycowithanolides, consisting of equimolar concentrations of sitoindosides VII-X and withaferin A, isolated from the roots of Withania somnifera Dunal, have been reported to have an antioxidant effect in the rat brain frontal cortex and striatum. In the present study, the effect of 10 days of oral administration of these active principles, in graded doses (10, 20 and 50 mg/kg), was noted on iron overload (FeSo(4), 30 mg/kg, i.p.) induced hepatotoxicity in rats. Apart from hepatic lipid peroxidation (LPO), the serum enzymes, alanine aminotransferase, aspartate aminotransferase and lactate dehydrogenase, were assessed as indices of hepatotoxicity. Silymarin (20 mg/kg, p.o.) was used for comparison. Iron overload induced marked increase in hepatic LPO and serum levels of the enzymes, which was attenuated by WSG in a dose-related manner, and by silymarin. The results indicate that the reported use of WS in Ayurveda for hepatoprotection against heavy metals and other environmental toxins, may be due the antioxidant action of WSG. PMID- 11054856 TI - Combined antiinfluenza virus activity of Flos verbasci infusion and amantadine derivatives. AB - The infusion prepared from flowers of Verbascum thapsiforme Schrad. (Scrophulariaceae) (FVI) reduced the infectious and haemagglutination yields of a range of influenza viruses in tissue cultures. Amantadine hydrochloride is an accepted and well studied selective inhibitor of influenza virus reproduction. The combined application of the plant preparation FVI and three amantadine derivatives resulted in a marked enhancement of the inhibitory effect of FVI on the reproduction of influenza virus A/chicken/Germany/27, strain Weybridge (H7N7) in cell cultures of chicken embryo fibroblasts. The antiviral activity was determined by the difference in the infectious titres of control and treated viruses. The combined effect was defined on the basis of infectious viral yields. The most pronounced enhancement was shown for the combination of FVI and adamantanamine glucuronide. PMID- 11054857 TI - Cytotoxicity and antibacterial activity of ethanol extract from leaves of a herbal drug, boneset (Eupatorium perfoliatum). AB - The cytotoxic and antibacterial activity of an ethanol extract of leaves of a herbal drug, boneset (Eupatorium perfoliatum), was investigated. The extract showed potent cytotoxicity with EC(50) values (12-14 microg/mL) comparable to a standard cytotoxic agent, chlorambucil. The extract showed a weak antibacterial activity against gram-positive test organisms (Staphylococcus aureus and Bacillus megaterium). PMID- 11054859 TI - Selected bibliography PMID- 11054858 TI - Patents alert PMID- 11054860 TI - Microscopy and the gut mucosa: into the next millennium. PMID- 11054861 TI - Altered enteroendocrine cell expression in T cell receptor alpha chain knock-out mice. AB - Mice lacking T cell receptor alpha chain (TCRalpha(-/-)) develop inflammation of the colon. We have examined the effect of this inflammation on the colonic epithelium by studying markers of epithelial cuff, enteroendocrine, and immune cell differentiation. Using immunohistochemical techniques, colons were compared in normal C57/BL6 and murine TCR alpha(-/-) mice aged 2 and 3 weeks and 3-11 months. TCR alpha(-/-) mice aged 3-11 months had histologic evidence of inflammation with increased expression of CD45, CD4+, CD8+, and B220+ cells and a decrease in expression of IgA+ cells. There was a decrease in the number of cholecystokinin, serotonin, and neurotensin enteroendocrine expressing cells in the colon of TCR alpha(-/-) mice. These changes were not present in 2-3-week-old suckling/weaning mice. In contrast, peptide tyrosine tyrosine (PYY), glucagon like peptide-1, and gastrin expression did not change and small intestinal enteroendocrine cells remained unaltered. The change in colonic enteroendocrine cell expression appears to be a specific response, since only a subset of these cells was altered, and the epithelium was intact by histologic analysis. The absence of functional T cells in TCR alpha(-/-) colon has a marked effect on differentiation of a specific subpopulation of enteroendocrine cells, prior to loss of integrity of the epithelium. PMID- 11054862 TI - Effect of tissue non-specific alkaline phosphatase in maintenance of structure of murine colon and stomach. AB - The gastrointestinal tract of mammals secretes a phospholipid-rich membrane that is enriched in alkaline phosphatase (AP) and surfactant proteins (surfactant-like particle, SLP). The production of this particle is stimulated in the small intestine by fat feeding and in cultured cells in vitro by transfection with intestinal alkaline phosphatase (IAP). To test whether tissue non-specific alkaline phosphatase (TNAP) was a factor in stimulating surfactant-like particle production in stomach and colon (tissues expressing TNAP), mice lacking this enzyme were studied. Mice were harvested at 8 days of life, when body weight of homozygous animals (TNAP -/-) was about half that of congenic controls (TNAP +/+) or heterozygotes (TNAP +/-), but before seizures had begun. No difference in content of the major SLP protein (65 kDa) by Western blotting or immunocytochemistry was seen in stomach or colon of TNAP -/- vs. TNAP +/+ animals, but the content was only about half in the IAP-expressing small bowel. Transmission electron microscopy of the TNAP -/- small bowel showed large dilated lysosomes and residual bodies. Colonocytes and gastric surface epithelial cells from the same animals showed mitochondria containing homogeneous dense inclusions, consistent with neutral lipid. In the underweight homozygous animals, there was a decrease in the neuronal content of submucosal ganglia in the jejunum and ileum and of myenteric ganglia in the jejunum of TNAP -/- animals. These findings suggest that (1) TNAP is not important in maintaining surfactant-like particle content of tissues that express TNAP, (2) normal fat absorption is important in maintaining SLP content in the small intestine, and (3) TNAP is important in the maintenance of some intestinal structures, and perhaps their function. PMID- 11054863 TI - Regulation of intestinal regeneration: new insights. AB - Intestinal regeneration is the process by which intestinal injury penetrating deep to the lamina propria heals. The regenerative process involves epithelial cell migration and proliferation, changes in cellular function, adaptation of subepithelial tissues, and contraction of the injured area. This requires interaction of multiple cell types. While many observations have been made about the process of regeneration, its regulation is not well understood. Previous studies, performed primarily in a serosal patch model, have identified many potential regulatory factors. These include location and size of the injury, other associated injury, e.g., resection, and a variety of agents that influence one or more of the primary processes involved. Epidermal growth factor (EGF), in particular, appears to play a role in many aspects of regeneration. Recent advances in the understanding of intestinal growth regulation have provided new insights into the regulation of intestinal regeneration. Developmental studies in genetically manipulated mice suggest a role for gene products not previously implicated in regeneration. The importance of apoptosis in growth regulation has recently been emphasized. Mesenchymal-epithelial interactions have gained greater appreciation. Finally, it has become clear that immune cells and cytokines are important factors in this process. Transforming growth factor-beta (TGFbeta) has been implicated as another important regulator of several of the processes involved in intestinal regulation. Improved understanding of the regulation of intestinal regeneration will lead to new therapeutic approaches to stimulate intestinal healing in the clinical setting. PMID- 11054864 TI - Epidermal growth factor is critical for intestinal adaptation following small bowel resection. AB - The loss of small intestinal mucosal surface area is a relatively common clinical situation seen in both the pediatric and adult population. The most frequent causes include mesenteric ischemia, trauma, inflammatory bowel disease, necrotizing enterocolitis, and volvulus. Following surgical resection, the remnant intestine compensates or adapts to the loss of native bowel by increasing its absorptive surface area and functional capacity. Unfortunately, many patients fail to adapt adequately, and are relegated to lifelong intravenous nutrition. Research into intestinal adaptation following small bowel resection (SBR) has evolved only recently from the gross and microscopic level to the biochemical and genetic level. As understanding of this process has increased, numerous therapeutic strategies to augment adaptation have been proposed. Epidermal growth factor (EGF) is an endogenous peptide that is secreted into the gastrointestinal tract and able to influence gut ontogeny, as well as mucosal healing. Early studies have demonstrated its ability to augment the adaptive process. Focusing on a murine model of massive intestinal loss, the morphological, structural, biochemical, and genetic changes that occur during the intestinal adaptive process will be reviewed. The role of EGF and its receptor as critical mediators of the adaptive process will be discussed. Additionally, the ability of EGF to augment intestinal proliferation and diminish programmed cell death (apoptosis) following SBR will be examined. Enhancing adaptation in a controlled manner may allow patients to transition off parenteral nutrition to enteral feeding and, thereby, normalize their lifestyle. PMID- 11054865 TI - Cytoskeleton as a target for injury in damaged intestinal epithelium. AB - This report summarizes the findings of a series of studies undertaken to discern the role of the cytoskeleton in intestinal injury and defense. Two established cell lines were used for these studies. IEC-6 cells (a rat intestinal cell line) were incubated in Eagle's minimal essential medium with and without 16, 16 dimethyl prostaglandin E(2) (dmPGE(2); 2.6 microM) for 15 minutes and subsequently incubated in medium containing 10% ethanol (EtOH). The effects on cell viability and the actin cytoskeleton were then determined. Using a similar protocol, Caco-2 cells (a human colonic cell line) were employed to assess the microtubule cytoskeleton under these conditions. In both cell lines, EtOH extensively disrupted the cytoskeletal component being evaluated coincident with adversely affecting cell viability. Pretreatment with dmPGE(2) increased cell viability and abolished the disruptive effects on both the actin and microtubule cytoskeleton in cells exposed to EtOH. Prior incubation with cytochalasin D, an actin disruptive agent, prevented the protective capabilities of dmPGE(2) in IEC 6 cells challenged with EtOH. Phalloidin, an actin stabilizing agent, demonstrated similar effects to that of dmPGE(2) by stabilizing the actin cytoskeleton and preserving cellular viability in IEC-6 cells in response to EtOH. In Caco-2 cells, taxol, a microtubule stabilizing agent, mimicked the effects of dmPGE(2) by increasing cell viability in cells exposed to EtOH and enhancing microtubular integrity. In contrast, pretreatment with colchicine, an inhibitor of microtubule integrity, prevented the protective effects of dmPGE(2). These findings support the hypothesis that the cytoskeleton may be a major target for injury in damaged intestinal epithelium, and that the protective action of dmPGE(2) is orchestrated through preservation of this target. PMID- 11054866 TI - Mechanism of extracellular calcium regulation of intestinal epithelial tight junction permeability: role of cytoskeletal involvement. AB - Recent studies suggest that an abnormal increase in intestinal tight junction (TJ) permeability may be an important etiologic factor in number of diseases including Crohn's disease, NSAID-associated enteritis, and various infectious diarrheal syndromes. The intracellular processes involved in regulation of intestinal epithelial TJ permeability, however, remain poorly understood. In this study, we used cultured Caco-2 intestinal epithelial cells to examine the intracellular processes involved in extracellular Ca(++) modulation of intestinal epithelial monolayer TJ barrier. Incubation of the filter-grown Caco-2 intestinal monolayers in Ca(++)-free solution (CFS), consisting of modified Krebs-buffer solution containing 0 mM Ca(++) and 1 mM EGTA, resulted in a rapid drop in Caco-2 epithelial resistance and increase in epithelial permeability to paracellular markers mannitol and inulin, indicating an increase in TJ permeability. The increase in Caco-2 TJ permeability was rapidly reversed by the re-introduction of Ca(++) (1.8 mM) into the incubation medium. The CFS-induced increase in Caco-2 TJ permeability was associated with separation of the cytoplasmic and transmembrane TJ proteins, ZO-1 and occludin, and formation of large intercellular openings between the adjoining cells. The CFS-induced modulation of TJ barrier was associated with activation of myosin light chain kinase (MLCK) activity and centripetal retraction of peri-junctional actin and myosin filaments. The inhibition of CFS-induced activation of Caco-2 MLCK with MLCK inhibitor (ML-7) prevented the CFS-induced retraction of actin and myosin filaments and the subsequent alteration of TJ barrier function and structure. Our results suggested that the CFS-induced alteration of TJ proteins and functional increase in TJ permeability was mediated by Caco-2 MLCK activation and the resultant contraction of the peri-junctionally located actin-myosin filaments. Consistent with the role of MLCK in this process, selected inhibitors of Mg(++)-myosin ATPase and metabolic energy, but not protein synthesis inhibitors, also prevented the CFS induced retraction of actin and myosin filaments and the subsequent increase in TJ permeability. In conclusion, our results indicate that extracellular Ca(++) is crucial for the maintenance of intestinal epithelial TJ barrier function. The removal of extracellular Ca(++) from the incubation medium causes activation of Caco-2 MLCK, which in turn leads to an increase in intestinal monolayer TJ permeability. PMID- 11054867 TI - Integrins as mediators of epithelial cell-matrix interactions in the human small intestinal mucosa. AB - The intestinal epithelium is a highly dynamic tissue, which depends on a variety of factors for the regulation of its rapid renewal and expression of digestive functions. Over the last 10 years, it has become evident that among these factors are cell interactions with the extracellular matrix, more specifically with the underlying basement membrane, through a series of specific cell membrane receptors, many of which are integrins. Integrins regulate the assembly of adhesive junctions as well as the activation of various signaling pathways, leading to the modulation of gene expression. The analysis of the integrin repertoire along the crypt-villus axis in the human small intestinal epithelium identifies a number of beta1 and beta4 integrins, showing differential patterns of expression relative to its two functional compartments. Among them are the integrins alpha3beta1, alpha7Bbeta1 and the functional form of alpha6beta4 that appear to be related, in concert with the distribution of their ligands, to the process of intestinal cell differentiation, and the integrins alpha2beta1, alpha1beta1, alpha5beta1, and the non-functional form of alpha6beta4 that seem to be coupled with the undifferentiated/proliferative status of crypt cells. These observations delineate the potential complexity of the organization of epithelial cell-matrix interactions involved in the maintenance of the human intestinal crypt-villus axis. PMID- 11054868 TI - Adhesion complexes implicated in intestinal epithelial cell-matrix interactions. AB - This article review summarizes data on cell-substratum adhesion complexes involved in the regulation of cellular functions in the intestine. We first focus on the molecular composition of the two main adhesion structures-the beta1 integrin-adhesion complex and the hemidesmosome-found in vivo and in two human intestinal cell lines. We also report the key findings on the cellular behavior and response to the extracellular matrix that involve integrins, the main transmembrane anchors of these complexes. How the dynamics of cell/extracellular matrix interactions contribute to cell migration, proliferation, differentiation, and tumorigenicity is discussed in the light of the data provided by the human intestinal cells. PMID- 11054869 TI - Matrix-specific FAK and MAPK reorganization during Caco-2 cell motility. AB - We have previously reported that Caco-2 cell motility redistributes FAK, paxillin, and activates p38. However, the subcellular organization of these intracellular signals during cell migration is unclear. We, therefore, investigated the organization of actin, FAK, paxillin, and activated ERK and activated p38 during Caco-2 motility across collagen I, fibronectin, laminin, and tissue culture treated glass. Differential density seeding generated homogeneous static and migrating populations. Expression of actin, FAK, paxillin, phospho ERK, and phospho-p38 were examined by immunofluorescent staining in static and motile cells. Actin was concentrated toward the peri-nuclear central area of cells migrating on matrix proteins studied. Actin immunoreactivity was decreased in the leading edge of lamellipodia. FAK immunoreactivity was weaker in migrating cells than in static cells on the same matrix. FAK was expressed along cell-cell contacts of both cell populations, but absent in migrating lamellipodia of matrix cultured cells. Paxillin staining was diffuse in static cells but organized toward migrating lamellipodia in a radial manner. Like FAK, phosphorylated ERK was expressed in the central region of migrating cells but was dramatically decreased at areas of cell-cell contact and free lamellipodia. Fibronectin exerted the greatest effect on ERK activation in all matrix proteins studied. In contrast, phosphorylated p38 staining was stronger in migrating cells on matrix than in static cells on the same matrix. Phosphorylated p38 was expressed in the nuclear of migrating cells and disappeared in the cell-cell contact side and free lamellipodia. Interestingly, the reorganization of these proteins was distinctly different on tissue culture treated glass without a physiologic matrix substrate. For instance, FAK staining increased rather than decreased in motile cells on plastic, and lamellipodial FAK staining could be discerned. Matrix may influence Caco-2 biology during migration not only by triggering intracellular phosphorylation events but also by reorganizing the cytoskeleton and the subcellular localization of these intracellular signals. PMID- 11054870 TI - Imaging of three-dimensional epithelial architecture and function in cultured CaCo2a monolayers with two-photon excitation microscopy. AB - The principal functions of the gastrointestinal tract mucosa include nutrient absorption, protein and fluid secretion, and the regulated symbiosis with intraluminal contents. Research in epithelial biology has benefited significantly from the use of cultured monolayer preparations, which closely replicate the structure and function of normal gastrointestinal mucosa. Given the explicit importance of epithelial architecture to its physiology, investigations of epithelial biology should be enhanced by the capacity to track microscopic structures and substances in live cells. In order to achieve this goal, it is necessary to employ a microscopic technique with the capability of imaging deep into the tissue or cell preparation, without adversely affecting its physiology. Two-photon excitation microscopy may constitute such a technique, due to its ability to provide fluorescence excitation of fluorophores using near infrared radiation, that has lower tissue absorption and scattering coefficients. This allows the efficient collection of light energy from sites hundreds of microns deep, with only minimal tissue damage. In this report, we have presented an introduction to the theory and practice of two-photon microscopy for imaging the GI tract epithelium, and have presented examples of its utility in discerning three-dimensional structure and function in CaCo2A epithelial cell monolayers. PMID- 11054871 TI - Laminins: an overview. PMID- 11054872 TI - Laminins: structure and genetic regulation. AB - The laminins form a large family of modular proteins found in basement membranes, but also elsewhere. They function as structural components and are essential for morphogenesis, but in addition interact with cell surface receptors such as integrins and alpha-dystroglycan. By virtue of their receptor interactions, they initiate intracellular signalling events that regulate cellular organization and differentiation. The many interactions of laminins are mediated by binding sites, often contributed by single domains, which may differ between different forms of laminin. In the present article, we describe how the diversity of laminins and the genetic regulation of the expression of different laminin forms lead to the formation of extracellular matrices with variable laminin composition and thereby different biological properties. PMID- 11054873 TI - Transcriptional regulation of laminin gene expression. AB - Laminins are the most abundant structural non-collagenous glycoproteins ubiquitously present in basement membranes. They are multidomain molecules consisting of of alpha, beta, and gamma chains. Although the precise functional differences between the laminin variants are not well understood, the diversity of laminin isoforms may reflect the formation of distinct basement membranes. The laminins display a remarkable restricted expression profile, suggesting a fine regulation of their genes. In this review, we focus on the most recent developments of laminin biology, centering on transcriptional and posttranscriptional controls. We discuss only those laminin chains whose gene organization and promoter elements have been characterized and proved to be functional. When possible, we correlate the effects of growth factors, cytokines, retinoids, and transcription factors on laminin gene expression with the identity of cis-acting elements in their genomic control regions. PMID- 11054874 TI - Proteolytic modification of laminins: functional consequences. AB - The laminin family contains a number of complex, multi-domain proteins that participate in a large variety of biologic processes. Limited proteolysis has been utilized extensively as a tool with which to determine laminin structure/function relationships. In addition, proteolytic modification of laminins may occur as a component of heterotrimer assembly and secretion, or may follow incorporation of mature laminin into the extracellular matrix. Conversely, laminin binding to cellular receptors may also influence proteinase expression. This review will highlight specific examples to demonstrate the functional interplay between laminins and proteinases in the regulation of laminin structure and function as well as in the subsequent control of proteinase expression. PMID- 11054875 TI - Laminins of the neuromuscular system. AB - The mammalian neuromuscular system expresses seven laminin genes (alpha 1, alpha 2, alpha 4, alpha 5, beta 1, beta 2, and gamma 1), produces seven isoforms of the laminin trimer (laminins 1, 2, 4, 8, 9, 10, and 11), and distributes these trimers to at least seven distinct basal laminae (perineurial, endoneurial, terminal Schwann cell, myotendinous junction, synaptic cleft, synaptic fold, and extrajunctional muscle). The patterns of expression, assembly, and distribution are regulated during development, and primary and secondary changes in laminin expression occur in several neuromuscular genetic disorders. Functional studies using knockout and transgenic mice, and purified laminins and cell types, demonstrate that laminins are required components of basal laminae in the neuromuscular system. Collectively, laminins have both structural and signaling functions; individually, laminin isoforms have unique roles in regulating the behavior of nerve, muscle, and Schwann cell. Among them, laminin-2 (alpha 2 beta 1 gamma 1) plays an important structural role in supporting the muscle plasma membrane, laminin-4 regulates adhesion and differentiation of the myotendinous junction, and laminin-11 regulates nerve terminal differentiation and Schwann cell motility. Together, these observations reveal remarkable diversity in the formation and function of laminins and basal laminae, and suggest avenues for addressing some neuromuscular diseases. PMID- 11054876 TI - Laminins and human disease. AB - The laminin protein family has diverse tissue expression patterns and is involved in the pathology of a number of organs, including skin, muscle, and nerve. In the skin, laminins 5 and 6 contribute to dermal-epidermal cohesion, and mutations in the constituent chains result in the blistering phenotype observed in patients with junctional epidermolysis bullosa (JEB). Allelic heterogeneity is observed in patients with JEB: mutations that results in premature stop codons produce a more severe phenotype than do missense mutations. Gene therapy approaches are currently being studied in the treatment of this disease. A blistering phenotype is also observed in patients with acquired cicatricial pemphigoid (CP). Autoantibodies targeted against laminins 5 and 6 destabilize epithelial adhesion and are pathogenic. In muscle cells, laminin alpha 2 is a component of the bridge that links the actin cytoskeleton to the extracellular matrix. In patients with laminin alpha 2 mutations, the bridge is disrupted and mature muscle cells apoptose. Congenital muscular dystrophy (CMD) results. The role of laminin in diseases of the nervous system is less well defined, but the extracellular protein has been shown to serve an important role in peripheral nerve regeneration. The adhesive molecule influences neurite outgrowth, neural differentiation, and synapse formation. The broad spatial distribution of laminin gene products suggests that laminin may be involved in a number of diseases for which pathogenic mechanisms are still being unraveled. PMID- 11054877 TI - Integrins as receptors for laminins. AB - Laminins are a family of trimeric glycoproteins present in the extracellular matrix and the major constituents of basement membranes. Integrins are alpha beta transmembrane receptors that play critical roles in both cell-matrix and cell cell adhesion. Several members of the integrin family, including alpha 1 beta 1, alpha 2 beta 1, alpha 3 beta 1, alpha 6 beta 1, alpha 7 beta 1 and alpha 6 beta 4 heterodimers serve as laminin receptors on a variety of cell types. This review summarizes recent advances in understanding the involvement of individual integrins in cell interactions with laminins and the roles of laminin-binding integrins in adhesion-mediated events in vertebrates, including embryonic development, cell migration and tumor cell invasiveness, cell proliferation and differentiation, as well as basement membrane assembly. We discuss the regulation of integrin function via alternative splicing of cytoplasmic domains of alpha and beta subunits of the integrin receptors for laminins and present examples of functional collaboration between laminin-binding integrins and non-integrin laminin receptors. Advances in our understanding of the laminin-binding integrins continue to demonstrate the essential roles these receptors play in maintaining cell polarity and tissue architecture. PMID- 11054878 TI - Proceedings of the 3rd International Symposium on the Diabetic Foot. 5-8 May 1999, Noordwijkerhout, The Netherlands. PMID- 11054879 TI - The diabetic foot: a global view. AB - It is estimated that approximately 15% of the more than 150 million people with diabetes world-wide will at some stage develop diabetic foot ulceration. Foot problems are indeed a global problem and there is no area in the world that does not report the development of foot lesions as a consequence mainly of neuropathy and peripheral vascular disease. The prevalence of active foot ulceration varies from approximately 1% in certain European and North American studies to more than 11% in reports from some African countries. Although there have been many developments in recent years which encourage optimism for future improvement in diabetic foot care, there is still much to be done; the recent data from the Netherlands show that with a concerted team approach, it is possible to increase the numbers of foot clinics with the provision of podiatry services by more than 100%. However, many countries still lack proper podiatry and specialist nursing provision and there remains much to be done in the next millennium to improve the lot of the diabetic patient with foot problems. PMID- 11054880 TI - New developments in the biomechanics of the diabetic foot. AB - Biomechanical issues are now widely recognized as being important in the treatment of diabetic foot disease. The purpose of the present review is to identify advances that have occurred since the previous International Conference on the Diabetic Foot in 1995 in the understanding of foot biomechanics in relation to diabetes. Attention continues to be focused on the identification of a threshold plantar pressure that leads to tissue damage. Recent studies have suggested that peak barefoot pressure may be only 65% specific for the development of ulceration. The association between foot deformity and plantar pressure has been the subject of several quantitative studies, but new questions have been raised about the etiology of claw toes. The measurement of shear stress continues to be an elusive goal although several small studies have presented possibly feasible technical approaches. The importance of callus as a precursor to ulceration has been confirmed experimentally and quantitative measures of motor neuropathy have been presented. Although a number of new devices have been introduced as alternatives to the Total Contact Cast, few clinical studies of their efficacy are available yet. New information on the properties of insole materials has been published including data on changes with repeated cycling. Complications of prophylactic surgery have been shown to include a high rate of Charcot fractures. Two new series describing the fixation of such fractures have also been reported. Biomechanical issues have also been addressed in two sets of guidelines for treatment that have recently been published. These many studies confirm the central role of mechanical stress and its relief in the treatment of neuropathic foot problems in diabetes. PMID- 11054881 TI - Peripheral vascular disease and type 2 diabetes mellitus. AB - Patients with Type 2 diabetes mellitus frequently have peripheral vascular disease, with a predilection for the lower legs. In this review potential mechanisms for this high prevalence and altered distribution are explored. It is hypothesised that the metabolic abnormalities in the prediabetic phase predispose to a more distal and aggressive atherosclerosis. Once diabetes has developed this process is accelerated due to chronic hyperglycaemia. Furthermore, endothelial damage, non-enzymatic glycosylation and polyneuropathy could lead to impaired vascular remodelling and collateral formation. PMID- 11054882 TI - Vascular imaging and intervention in peripheral arteries in the diabetic patient. AB - Diabetic patients are four times more likely to develop peripheral vascular disease than the general population. This disease is likely to be more aggressive, with five times more patients developing critical limb ischaemia. Early diagnosis and treatment allows up to 80% of these patients to have some form of surgical or endovascular re-vascularisation. The primary imaging modalities to be used should be duplex ultrasound followed by angiography. Magnetic resonance angiography, however, holds out promise for the future as being a good method of non-invasive imaging. Endovascular (interventional radiological) procedures have a major role to play in treatment of vascular stenoses and occlusions. Thrombolytic agents can be used to dissolve thrombus within occluded vessels and so restore patency. Percutaneous transluminal angioplasty is of value in dilating the stenotic lesions within the vessels and so restoring normal blood flow. Endovascular stents may be inserted to ensure longer term patency. There is indirect evidence to suggest that the outcomes of endovascular procedures in the diabetic patient are less good than those in the general population, but nevertheless such procedures may save the diabetic patient from primary amputation and allow healing of ischaemic ulcers. PMID- 11054883 TI - Foot debridement: anatomic knowledge is mandatory. AB - The foot is well adapted for its purpose. This is reflected by its special anatomical architecture, having a thickened epidermis, subcutaneous fatty pads and muscles arranged in compartments. In diabetic patients, metabolic changes and foot deformity can lead to high pressure zones resulting in a foot ulcer. If an ulcer gets infected, the oedema aggravates the underlying muscle compartment pressure, even leading to muscle necrosis. This explains why foot ulcers can lead to extensive tissue necrosis. For debridement, sound anatomical knowledge of the foot anatomy is mandatory. PMID- 11054884 TI - Never amputate without consultation of a vascular surgeon. AB - Lower limb ischaemia is one of the determinants in the development of diabetic foot ulcers and the most important factor preventing their healing. There are a number of misleading factors masking the presence of atherosclerotic disease and tissue damage; these are reduced inflammatory response to infection, autosympathectomy and mediasclerosis, which all diminish the clinical suspicion of ischaemia. Therefore, adequate assessment of the lower limb circulation should be routinely performed in complicated diabetic foot. This evaluation can often be made with simple methods. In addition to clinical examination ankle/brachial pressure index, systolic toe pressure, plethysmographic pulse volume recordings and simple hand-held Doppler auscultation are most often sufficient to make a decision as to whether angiography is needed or not. Duplex examination can give more profound information on the severity and extent of arterial occlusive disease, but the method is strongly user-dependent. Early vascular consultation is mandatory in diabetic foot work-up and should be undertaken within 2 weeks if a new skin lesion shows no tendency to heal. Long bypass grafting procedures and microvascular free flap techniques have been shown to achieve excellent results in relieving critical leg ischaemia, even in the presence of large foot lesions, and should be used to prevent major amputation. The timing of various procedures is a controversial issue. Feet with small ulcers or restricted dry gangrena can be revascularised first, with minor amputations and local surgery of the ulcer being done thereafter. In the septic neuroischaemic foot, major amputation may be unavailable but if the infection is not immediately life-threatening the infected part of the foot should be drained and debrided properly and left wide open, sometimes with a guillotine amputation in order not to risk the bypass graft, which can be done a couple of days later. PMID- 11054885 TI - Combined vascular reconstruction and free flap transfer in diabetic arterial disease. AB - Gangrenous lesions of the foot or lower leg due to severe diabetic arterial disease resulting in extensive soft tissue defects with exposed bones or tendons often result, even after successful revascularisation, in staged or primary amputation. We present our experience with 45 such patients treated with combined arterial reconstruction and free tissue transfer for limb-salvage. All presented with peripheral vascular disease of diabetic origin, and extensive gangrenous lesions that could not be treated by simple wound closure or skin-grafting without major amputation. A total of 53 arterial reconstructions and 47 free-flap transfers were performed. In the majority of patients, the distal anastomosis was on a pedal or tibial vessel. These bypass grafts or a native revascularized artery served as the inflow tract for the free flap which was anastomosed using microsurgical techniques. Venous anastomoses were preferentially performed on the deep venous system. Donor muscles were Musculus rectus abdominis (n=37), Musculus latissimus dorsi (n=5), Musculus serratus anterior (n=3), and a perforator flap (n=2) tailored to the size of the defect and covered with a split thickness graft (STG). The operation was set up as a combined procedure in 39/45 patients, two teams working simultaneously, limiting the mean operative time to 6 h. Early reinterventions had to be performed in 14 patients resulting in five flap losses of which two could be treated with a new free flap transfer and three were amputated. Three other patients died in the postoperative period, leaving us with a total of 39/45 patients leaving the hospital with a full-length limb. Independent ambulation was achieved in 32 of these 39 patients. During late follow-up (mean 26 months) eight bypasses occluded resulting in two amputations and two new vascular reconstructions. Combined survival and limb-salvage rate was 84% after 1 year, 77% after 2 years and 65% after 3 years. The advantages of this combined technique are: (1) it provides immediate soft tissue coverage limiting amputation level and healing time, resulting in early ambulation; (2) it provides extra run-off to the revascularisation, illustrated by a decrease in peripheral resistance, contributing to its patency; (3) the application of healthy, well vascularised tissue limits infection and enhances neovascularisation; (4) a full length limb is preserved. We believe this combined approach offers a valuable alternative to primary amputation in this group of patients with extensive ischaemic defects. PMID- 11054886 TI - Pharmacotherapy for critical limb ischaemia. AB - Critical limb ischaemia involves a severe disturbance of both macrocirculation and microcirculation. Provided revascularisation can be performed (balloon angioplasty, bypass procedures or other reopenings), the major pharmacological problem is to reduce the risk of early thrombosis and midterm hyperplastic growth. Adjuvant treatment for the former is useful, while the latter has not yet found any specific solution. When no technical prerequisites exist for revascularisation, which means extremely deranged vascular morphology, pharmacotherapy can be tried to reduce the specific micro-circulatory events. Results are far from good, though significant reduction of amputations and deaths may be achieved with certain drugs. PMID- 11054887 TI - Principles and practice of antibiotic therapy of diabetic foot infections. AB - Foot infections are a common and serious problem in diabetic patients. They usually occur as a consequence of a skin ulceration, which initially is colonized with normal flora, and later infected with pathogens. Infection is defined clinically by evidence of inflammation, and appropriate cultures can determine the microbial etiology. Aerobic gram-positive cocci are the most important pathogens; in chronic, complex or previously treated wounds, gram-negative bacilli and anaerobes may join in a polymicrobial infection. In all diabetic foot infections a primary consideration is whether or not surgical intervention is required, e.g. for undrained pus, wound debridement or revascularization. Antibiotic regimens are usually selected empirically initially, then modified if needed based on results of culture and sensitivity tests and the patient's clinical response. Initial therapy, especially in serious infections, may need to be broad-spectrum, but definitive therapy can often be more targeted. Severe infections usually require intravenous therapy initially, but milder cases can be treated with oral agents. Treatment duration ranges from 1-2 weeks (for mild soft tissue infection) to more than 6 weeks (for osteomyelitis). The choice of a specific agent should be based on the usual microbiology of these infections, data from published clinical trials, the severity of the patient's infection, and the culture results. Extension of infection into underlying bone can be difficult to diagnose and may require imaging tests, e.g. magnetic resonance scans. Cure of osteomyelitis usually requires resection of infected bone, but can be accomplished with prolonged antibiotic therapy. Various non-antimicrobial adjunct therapies may sometimes be helpful. Published in 2000 by John Wiley & Sons, Ltd. PMID- 11054888 TI - Topical treatment: which dressing to choose. AB - Wounds have existed since the beginning of time. The interest in this subject has been stimulated in the main by conflict and war that have necessitated the development of new ways of managing wounds. In the 1960s the development of new materials that maintained a moist environment in the wound area encouraged a number of commercial companies to produce a wide variety of new materials with physical and chemical properties that might provide a moist environment. However the data to support the use of such materials are limited if one requires evidence that they have produced more rapid healing in chronic wounds kept moist as to those kept dry. Is this due to a problem with the outcome measure rather than a problem with the materials themselves? Rather than seeing this as justification for not using such materials, it should instead lead clinicians to question the validity of endpoint studies in wound healing experiments. There is a lack of evidence regarding the ability of such materials to improve the speed of healing in chronic wounds. Nevertheless considerable clinical experience, obtained from treating many patients, has indicated that not only are such new treatments cost effective, but that they are also proving to be extremely beneficial and acceptable to patients, on account of their ability to reduce pain, odour or leakage from a wound. PMID- 11054889 TI - New treatments in ulcer healing and wound infection. AB - This review examines several of the recently introduced wound care products that have been put forward as treatment modalities for the diabetic foot ulcer. Discussed are the results of clinical trials with the platelet-derived growth factor, becaplermin, the tissue-engineered products Dermagraft and Apligraf, and Hyaff which is an ester of hyaluronic acid. In patients with an infected foot ulcer, encouraging results were obtained with the granulocyte-colony stimulating factor, Filgrastim. PMID- 11054890 TI - Hyperbaric oxygen therapy and the diabetic foot. AB - Common causes for non-healing of diabetic foot ulcers are infection and/or ischaemia. Diabetic patients are compromised hosts as far as wound healing is concerned. Diabetes mellitus is associated with a defective cellular and humoral immunity. In particular, decreased chemotaxis, decreased phagocytosis, impaired bacterial killing and abnormal lymphocytic function have been observed, resulting in a reduced inflammatory reaction and defective wound healing. The potential benefits of hyperbaric oxygen therapy (HBO) in diabetic patients with a foot ulcer are discussed. Oxygen plays an important role in the physiology of wound healing. HBO can raise tissue oxygen tensions to levels where wound healing can be expected. Hyperbaric oxygen increases also the killing ability of leucocytes, is lethal for certain anaerobic bacteria and inhibits toxin formation in other anaerobes. Multiple anecdotal reports and retrospective studies in HBO therapy in diabetic patients suggest that HBO can be an effective adjunct in the management of diabetic wounds. Prospective studies also show the beneficial effects of HBO. Because most published studies suffer from methodological problems, there is an urgent need for a collaborative, international, randomised prospective clinical trial for the application of HBO in diabetic foot lesions, as part of a multidisciplinary treatment approach, before we can recommend HBO as standard therapy in patients with foot ulcers. PMID- 11054891 TI - Management of the diabetic Charcot foot. AB - The diabetic Charcot foot is a major limb-threatening complication of long-term diabetes mellitus and neuropathy. Although first described over 100 years ago, we are still lacking definitive studies regarding its prevalence in this population, precise etiology, or most effective treatments. Trauma in the presence of peripheral sensory neuropathy and abundant arterial perfusion seem to be the primary causal factors leading to this severe foot deformity. Misdiagnosis or delayed diagnosis of osteoarthropathy allows the destructive phase of this disorder to continue with resultant further destruction of the foot architecture. The authors discuss the natural history of this entity as well as potential treatment options and recommendations. Through a better understanding of the underlying pathogenesis, Charcot arthropathy can be more effectively managed and thereby limit the development of severe deformity, ulceration, infection and limb loss. PMID- 11054892 TI - Surgical treatment of the diabetic foot. AB - The majority of patients with diabetic foot problems can be treated non surgically. In this article a short overview is given of the surgical procedures employed in the salvage of the diabetic foot with discussion of the indications and techniques. PMID- 11054893 TI - Diabetic patient education: determinants of success. AB - Education/empowerment is critical if successful self-management is to be achieved. All professional patient interactions have a learning component. Clinical outcomes in diabetes are as dependent on psycho-social factors or learned behaviour as on metabolic state or therapeutic interventions. These factors include targets set, self-management skills, influence of living with diabetes, emotional factors, role of other people, perceived benefits and barriers, feelings of self-efficacy, weight concern and diet barrier. Training in learning processes and factors governing behaviour are essential for all those involved in delivery of patient care. Educational programmes should recognise the wide range of learning strategies used by different people. PMID- 11054894 TI - What is the most effective way to reduce incidence of amputation in the diabetic foot? AB - Approximately 40-60% of all amputations of the lower extremity are performed in patients with diabetes. More than 85% of these amputations are precipitated by a foot ulcer deteriorating to deep infection or gangrene. The prevalence of diabetic foot ulcers has been estimated to be 3-8%. The complexity of these ulcers necessitates a multifactorial approach in which aggressive management of infection and ischemia is of major importance. For the same reason, a process oriented approach in the evaluation of prevention and management of the diabetic foot is essential. Healing rates of foot ulcers are unknown with the exception of specialised centres where it is between 80-90%. The negative consequences of diabetic foot ulcers on quality of life include not only morbidity but also disability and premature mortality. Costs for healing ulcers are high and even higher for ulcers resulting in amputation, due to prolonged hospitalisation, rehabilitation, and need for home care and social service for disabled patients. Therefore, one of the most important steps to reduce cost in the management of the diabetic foot is to avoid amputations. A cost-effective management should not only be focused on the short-term cost until healing but also on the long-term cost, since foot ulcer and especially amputation are related to increased re ulceration rate and lifelong disability. A multidisciplinary approach including preventive strategy, patient and staff education, and multifactorial treatment of foot ulcers has been reported to reduce the amputation rate by more than 50%. PMID- 11054895 TI - International consensus and practical guidelines on the management and the prevention of the diabetic foot. International Working Group on the Diabetic Foot. AB - In 1999 the International Consensus on the Diabetic Foot was published by a group of independent experts. The consensus process is described in this article together with the Practical Guidelines which were part of the consensus document. PMID- 11054896 TI - Pressure support and pressure assist/control: are there differences? An evaluation of the newest intensive care unit ventilators. AB - BACKGROUND: Pressure support (PS) has been widely studied in both patients and lung models, but there is little data available evaluating pressure assist/control (P A/C, frequently referred to as PCV) and no data comparing the operational capabilities of these two modes on the newest generation of ICU ventilators. We used a spontaneously breathing lung model to evaluate the response of the following new generation ventilators to varying inspiratory demand in both PS and P A/C: Bear 1000, Drager Evita 4, Hamilton Galileo, Nellcor Puritan-Bennett 840 and 740, Siemens Servo 300A, TBird AVS. METHODS: A bellows-in a-box lung model was set at a respiratory rate of 12 breaths/min, inspiratory time of 1.0 second, and peak inspiratory flows (modified square wave) of 40, 60, and 80 L/min. Each ventilator was set at three levels of PS and P A/C: 10, 15, and 20 cm H(2)O. On all ventilators, flow-triggering was set as sensitive as possible without causing self-triggering. RESULTS: Trigger pressure, trigger pressure-time product, inspiratory trigger time delay, ventilator-delivered peak flow, inspiratory area as a percent of the ideal inspiratory area, expiratory time delay, supraplateau expiratory pressure change, and expiratory area all varied among ventilators and at different lung model peak flows (p < 0.01 and >/= 10% difference). However, PS and P A/C on a given ventilator only differed with regard to expiratory variables (p < 0. 01 and >/= 10% difference). CONCLUSION: In a given ventilator little difference exists in gas delivery and response variables between PS and P A/C, but performance differences do exist among the ventilators evaluated. Ventilator performance is diminished at high lung model peak flows and low pressure settings. (I)), whereas PS gives control over ending inspiration to the patient. What has not been clearly defined is the gas delivery and ventilator response differences, if any, between these two (PS and P A/C) pressure targeted assist modes. Most new generation intensive care unit (ICU) ventilators provide both pressure support (PS) and pressure assist/control (P A/C) ventilation.19,20 The specific operational difference between these two modes is the mechanism that transitions inspiration to expiration. With pressure support the primary mechanism is a decrease in peak inspiratory flow to a predetermined level, whereas with P A/C mechanical T(I) is preset.19,20 We compared the operation of seven of the newest generation ICU ventilators in a spontaneously breathing lung model in both PS and P A/C. We hypothesized that there would be no difference in variables assessed between PS and P A/C except for the transition to expiration and that there would be no difference in response among ventilators evaluated. PMID- 11054897 TI - Inhaler technique of outpatients in the home. AB - OBJECTIVE: Assess the role of evaluation, instruction, and use of spacers by patients using metered dose inhalers (MDIs) in the home. PATIENTS AND SETTING: Patients (age 64 +/- 15 y [mean +/- standard deviation]) receiving home visits by respiratory care practitioners for oxygen therapy had their inhaler technique checked. INTERVENTIONS AND MEASUREMENTS: A detailed acceptable/unacceptable check off list was used with 172 patients to evaluate inhaler technique. Patients with poor technique were given instruction and their technique was reassessed. A subgroup of 43 patients was reevaluated on up to 3 visits. RESULTS: Only 18% of patients using MDIs without spacers were rated acceptable with the detailed check list. Instruction improved inhaler technique, but few patients with initially poor technique without spacers developed fully acceptable technique. Improvements made immediately following instruction were lost when patients were reevaluated months later. Few patients received spacers after they were recommended. Technique was markedly better with spacers. Most patients (76%) had initially proper technique with spacers, and most who had poor technique could learn and retain proper technique. CONCLUSIONS: Improper inhaler technique without spacers is very common among patients evaluated at home, and the majority of patients were unable to learn and retain proper technique. Most patients would benefit from using spacers with their inhalers. = 33 for first and second. = 26 for third. FRC = functional residual capacity. RV = residual volume. TLC = total lung capacity.?., Fig. 3?/PICK;0404f3;;;page;;;;yes;1? ?/GRAPH;rk1000404003;comptd;;center;stack? ?/CAPT;;;center;stack;2112n? PMID- 11054898 TI - Congenital cystic adenomatoid malformation in the newborn: two case studies and review of the literature. AB - Congenital cystic adenomatoid malformation (CCAM) is a congenital malformation of the lung that can present on imaging studies as abnormal air, air/fluid-filled cysts, or fluid-filled/solid-appearing cysts. The use of ultrasound in prenatal management has increased the number of cases diagnosed in utero. Early diagnosis is vital in the medical management of CCAM. Outcome varies from hydrops and fetal demise to complete resolution before birth. Many CCAMs diagnosed in utero may decrease in size even if substantial mediastinal shift and lung compression are noted at the time of diagnosis. Once the disorder has been diagnosed, use of serial ultrasound is helpful in providing medical management of the fetus. Two cases of CCAM in the newborn are presented that reflect characteristic clinical features but with distinctly different outcomes, one patient successfully responding to resection and ventilatory support, the other succumbing in the first day of life. The embryology, histology, prenatal and postnatal clinical presentation, and treatment of this malformation are discussed on the basis of a review of the literature. Recent developments in fetal diagnosis and treatment, including fetal surgery, are also presented. We conclude that CCAM should be considered in the differential diagnosis in the presence of respiratory distress and mediastinal shift. It is especially important for respiratory therapists, nurses, and other members of neonatal transport teams to consider CCAM in the differential diagnosis for any patient who presents with respiratory distress and a chest radiograph showing mediastinal shift. The treatment of choice for this lesion is surgical resection by either segmentectomy or lobectomy. Even in cases of relatively asymptomatic patients with CCAM, surgical resection should be considered because of the reported association of carcinoma and unresected CCAM. (IO2)) of 1.0 at a rate of 60-100 breaths per minute, there was slight improvement in oxygen saturation, to 50-70%. The postintubation radiograph showed bilateral haziness, with a mediastinal shift to the right. An umbilical venous catheter was placed and a venous blood gas study revealed a pH of 6.82, partial pressure of carbon dioxide of 100 mm Hg, and partial pressure of oxygen of 36 mm Hg. A needle aspiration of the left chest wall was performed to relieve the mediastinal shift; it produced approximately 40 mL of clear fluid. A repeat chest radiograph showed a possible pneumothorax on the left and continued mediastinal shift. A left-side chest tube was inserted and clear fluid continued to drain, but an additional chest radiograph was unchanged. Needle aspiration was then performed on the left side, and a large amount of air was removed. The S(pO2) increased to 88%, and the heart rate and blood pressure remained stable. The patient was then prepared for transport and placed on a Biomed MPV 10 (Bio-Med Devices, Guilford, Connecticut), intermittent mandatory ventilation rate 120, peak pressure 25 cm H(2)O, positive end-expiratory pressure 5 cm H(2)O (25/5), and F(IO2) 1.0. A venous blood gas study just before transport showed a pH of 7.1 and a P(CO2) of 45 mm Hg. The S(pO2) at that time was between 60% and 70%. To facilitate ventilation during transport, the patient was also given pancuronium (Pavulon) and morphine. On arrival at the neonatal intensive care unit, the patient was placed on a Sensormedics High-Frequency Oscillator (Sensormedics Corporation, Yorba Linda, California) with an initial amplitude of 70, airway pressure 25 cm H(2)O, inspiratory time 33%, Hertz 13, and F(IO2) 1.0. Three additional chest tubes were placed in the left hemithorax, which initially evacuated air, followed by serosanguineous fluid. The S(pO2) briefly increased to above 90%. A repeat chest radiograph again showed persistence of the left-sided collection of air and mediastinal shift. (ABSTRACT TRUNCATED) PMID- 11054899 TI - US public health service clinical practice guideline: treating tobacco use and dependence. AB - Treating Tobacco Use and Dependence, a Public Health Service-sponsored Clinical Practice Guideline, is a product of the Tobacco Use and Dependence Guideline Panel ("the panel"), consortium representatives, consultants, and staff. These 30 individuals were charged with the responsibility of identifying effective, experimentally validated tobacco dependence treatments and practices. The updated guideline was sponsored by a consortium of seven Federal Government and nonprofit organizations: the Agency for Healthcare Research and Quality (AHRQ), Centers for Disease Control and Prevention (CDC), National Cancer Institute (NCI), National Heart, Lung, and Blood Institute, National Institute on Drug Abuse, Robert Wood Johnson Foundation, and University of Wisconsin Medical School's Center for Tobacco Research and Intervention. This guideline is an updated version of the 1996 Smoking Cessation Clinical Practice Guideline No. 18 that was sponsored by the Agency for Health Care Policy and Research (now the AHRQ), United States Department of Health and Human Services. The original guideline reflected the extant scientific research literature published between 1975 and 1994. The updated guideline was written because new, effective clinical treatments for tobacco dependence have been identified since 1994. The accelerating pace of tobacco research that prompted the update is reflected in the fact that 3,000 articles on tobacco were identified as published between 1975 and 1994, contributing to the original guideline. Another 3,000 were published between 1995 and 1999 and contributed to the updated guideline. These 6,000 articles were screened and reviewed to identify a much smaller group of articles that served as the basis for guideline data analyses and panel opinion. This guideline contains strategies and recommendations designed to assist clinicians, tobacco dependence treatment specialists, and health care administrators, insurers, and purchasers in delivering and supporting effective treatments for tobacco use and dependence. The recommendations were made as a result of a systematic review and analysis of the extant scientific literature, using meta-analysis as the primary analytic technique. The strength of evidence that served as the basis for each recommendation is clearly indicated in the guideline. A draft of the guideline was peer-reviewed prior to publication, and the comments of 70 external reviewers were incorporated into the final document. The key recommendations of the updated guideline, Treating Tobacco Use and Dependence, based on the literature review and expert panel opinion, are as follows: 1. Tobacco dependence is a chronic condition that often requires repeated intervention. However, effective treatments exist that can produce long-term or even permanent abstinence. 2. Because effective tobacco dependence treatments are available, every patient who uses tobacco should be offered at least one of these treatments: Patients willing to try to quit tobacco use should be provided with treatments identified as effective in this guideline. Patients unwilling to try to quit tobacco use should be provided with a brief intervention designed to increase their motivation to quit. 3. It is essential that clinicians and health care delivery systems (including administrators, insurers, and purchasers) institutionalize the consistent identification, documentation, and treatment of every tobacco user seen in a health care setting. 4. Brief tobacco dependence treatment is effective, and every patient who uses tobacco should be offered at least brief treatment. 5. There is a strong dose-response relation between the intensity of tobacco dependence counseling and its effectiveness. Treatments involving person to-person contact (via individual, group, or proactive telephone counseling) are consistently effective, and their effectiveness increases with treatment intensity (eg, minutes of contact). 6. (ABSTRACT TRUNCATED) PMID- 11054900 TI - Intraretinal axon diameter: a single cell analysis in the marmoset (Callithrix jacchus). AB - We have labelled individual retinal ganglion cells of a New World primate, the common marmoset (Callithrix jacchus) with neurobiotin and then measured axon, soma and dendritic field diameter. A total of 111 cells were analysed (62 parasol cells, 22 midget cells, 16 hedge cells and 11 small bistratified cells). When all retinal ganglion cells were grouped together axon diameter was positively correlated to soma diameter. When analysed according to cell class only midget cells showed a positive correlation between soma size and mean axon diameter. Dendritic field diameter and mean axon diameter of both parasol and midget cells showed significant correlations. Axon diameter is not constant along the intraretinal length of the axon and the rate of change in diameter appears to be related to the cell class and the initial size of the axon. Midget cell axons showed a rapid increase of up to 20% over the first 200 microm in contrast to parasol cell axons which increased more slowly over this distance but then showed a marked increase in diameter of up to 40% over the next 450 microm. However, axon diameter did not remain at these increased diameters but decreased at greater distances from the soma. The degree to which an axon changes its diameter is related to retinal ganglion cell class and the initial size of the axon. We postulate that these variations in intraretinal axon diameter may have a direct influence on conduction velocity and reflect a compensatory mechanism to minimise spatiotemporal dispersion along the visual pathway. PMID- 11054901 TI - A longitudinal study of the effects of prenatal ethanol exposure on neuronal acquisition and death in the principal sensory nucleus of the trigeminal nerve: interaction with changes induced by transection of the infraorbital nerve. AB - The present study determines (1) whether ethanol-induced microencephaly results from reductions in neuronal acquisition (i.e., cell proliferation and neuronal migration) and/or increases in neuronal death and (2) whether ethanol exacerbates death by the same mode as that for naturally occurring or lesion-induced neuronal death. Pregnant rats were exposed to a diet containing 6.7% (v/v) ethanol or an isocaloric control diet during the last two weeks of gestation. At birth, the right infraorbital nerves of the pups were transected. The numbers of neurons in the principal sensory nucleus of the trigeminal nerve (PSN) on both sides of the pons were examined at various prenatal and early postnatal timepoints. The numbers of pyknotic and argyrophilic PSN cells were also counted. Ethanol delayed and reduced (19.9%) the prenatal acquisition of PSN neurons. The postnatal decline in neuronal number (indicative of neuronal death) was significantly increased (10.6%) by ethanol. Likewise, the numbers of pyknotic and silver stained cells were significantly higher in ethanol-treated rats. Lesion of the infraorbital nerve induced significant transsynaptic neuronal death in the control rats. Ethanol increased the amount of death caused by the lesion; however, it altered neither the timing of the neuronal loss nor the incidence of pyknosis or silver-staining. Therefore, ethanol affects both neuronal acquisition and survival; the greater effect being on neuronal acquisition. The timing and morphology of dying cells indicate that regardless of the cause (natural processes, ethanol-induced, or lesion-induced), neurons die in the developing PSN by the same mode. PMID- 11054902 TI - Cytoskeletal integration in a highly ordered sensory epithelium in the organ of Corti: reponse to loss of cell partners in the Bronx waltzer mouse. AB - This report is concerned with control of cell shaping, positioning, and cytoskeletal integration in a highly ordered cochlear neuroepithelium. It is largely based on investigations of events that occur during abnormal morphogenesis of the organ of Corti in the Bronx waltzer (bv/bv) mutant mouse. The organ's sensory hair cells and adjacent supporting cells ordinarily construct a spatially elaborate and supracellularly integrated cytoskeletal framework. Large microtubule bundles are connected to cytoskeletal components in neighbouring cells by actin-containing meshworks that link them to substantial arrays of adherens junctions. In bv/bv mice, degeneration and loss of most inner hair cells and outer pillar cells occurs during organ development. These cells flank each side of a row of inner pillar cells that respond by upregulating assembly of their actin-containing meshworks. This only occurs in surface regions where they no longer contact cell types involved in construction of the cytoskeletal framework. The meshworks are larger and exhibit a more extensive sub surface deployment than is normally the case. Hence, assembly of intercellular cytoskeletal connecting components can proceed without contact with appropriate cell neighbours but termination of assembly is apparently subject to a negative feedback control triggered by successful completion of intercellular connection with the correct cell neighbours. In addition, inner pillar cells compensate for loss of cell neighbours by interdigitating and overlapping each other more extensively than is usually the case to increase opportunities for generating adherens junctions. Certain adherens junctions in the organs of +/+ and bv/bv mice exhibit features that distinguish them from all previously described cell junctions. The dense plaques on their cytoplasmic faces are composed of aligned ridges. We suggest that they are called ribbed adherens junctions. Perturbations of cell shaping and positioning indicate that loss of inner hair cells is the primary consequence of the bv mutation. Most of the other abnormalities can be understood in terms of a secondary sequence of morphogenetic aberrations (precipitated by loss of inner hair cells). These aberrations provide new information about the ways in which supporting cells help to control hair cell positioning. PMID- 11054903 TI - Monoclonal antibody 2G13, a new axonal growth cone marker. AB - Growth cones are specialized sensorimotor structures at the tips of neurites implicated in pathfinding decisions and axonal outgrowth during neuronal development. We generated a mouse monoclonal antibody (mAb 2G13) against chick tectum and found that the antibody exclusively labelled axonal growth cones, particularly their filopodia and lamellipodia, in developing rat CNS and in embryonic neurons in culture. The high fidelity of the staining of growth cones by mAb 2G13 means that the antibody will be a useful marker for identifying growth cones. In growth cones of cultured neurons, mAb 2G13 labelling is intracellular and mainly associated with the filamentous actin cytoskeleton. Experiments with cytochalasins, which depolymerise filamentous actin, showed that 2G13p (the protein recognised by mAb 2G13) is physically associated with filamentous actin in growth cones. These properties of 2G13p suggest a role in growth cone motility. PMID- 11054904 TI - A light and electron microscopic study of the CB1 cannabinoid receptor in monkey basal forebrain. AB - The distribution of the CB1 cannabinoid receptor was studied in the monkey basal forebrain by immunocytochemistry and electron microscopy, using an antibody to the CB1 brain cannabinoid receptor. Large numbers of labelled neurons were observed in the medial septum, nucleus of the diagonal band, and the nucleus basalis of Meynert. The labelled neurons had dimensions similar to those of cholinergic neurons and were larger than those of GABAergic neurons. Double immunolabelling with an antibody to the synthetic enzyme for acetylcholine, choline acetyl transferase (ChAT) showed that CB1-positive neurons were also positive for ChAT, whilst electron microscopy confirmed that CB1-labelled neurons contained lipofuscin granules and dense clusters of rough endoplasmic reticulum, characteristic of cholinergic neurons. The dense labelling of cholinergic neurons for CB1 is interesting from the standpoint of neuroprotection. The CB1 receptor has been shown to couple in an inhibitory manner to voltage dependent calcium channels, and the dense labelling of CB1 in cholinergic neurons would therefore suggest that CB1 receptors could be important in limiting calcium influx through voltage dependent calcium channels in these neurons. This could serve to limit intracellular calcium concentrations, and consequent calcium mediated injury, in these neurons. PMID- 11054906 TI - Fenfluramines and Heart Valves. PMID- 11054905 TI - A light and electron microscopic study of GAT-1 in the monkey basal ganglia. AB - The distribution of the GABA transporter GAT-1 was studied by immunocytochemistry and electron microscopy in the monkey basal ganglia. Dense staining was observed in the globus pallidus externa and interna, intermediate in the subthalamic nucleus, and substantia nigra, and light staining in the caudate nucleus and putamen. Staining was observed in axon terminals, but not cell bodies. Electron microscopy showed that the GAT-1 positive axon terminals formed symmetrical synapses, suggesting that they were the terminals of GABAergic neurons. Comparison of areas high in GAT-1 protein with that of GABA showed a good correlation between the density in neuropil staining for GAT-1, and that of GABA. PMID- 11054907 TI - Introduction. Intracoronary thrombosis is the largest single cause of morbidity and mortality in the Western World. PMID- 11054908 TI - Pathophysiology of coronary thrombosis. AB - Detailed knowledge of the pathophysiology as well as the dynamic nature of coronary thrombus formation provides a valuable tool for correct management and proper adjunctive therapy in patients with acute coronary syndromes. Coronary thrombosis is in the majority of cases caused by disruption or fissuring of an atherosclerotic plaque. At the lesion thrombogenic material will be exposed to the flowing blood leading to activation of platelets and the formation of a platelet clot. Simultaneously, the coagulation system is activated resulting in increased thrombin formation. Thrombin is a key mediator in arterial thrombosis, due to its effect on both platelets and fibrin generation. Thrombin contributes to the stabilization of an initially loose platelet clot by generating cross bound fibrin within the thrombus. During the course of an acute coronary syndrome, the patient presents changing chest pain and dynamic ischaemic ECG findings. This is likely to be related to the dynamic nature of the pathophysiology. The presence of a non-occlusive coronary thrombus may deprive the myocardium its normal blood flow and oxygen supply, leading to ischaemic pain. During lysis or embolization, blood supply may be restored, but the presence of thrombus fragments in the microcirculation holds the potential to sustained interference with myocardial metabolism. The emboli contain activated platelets which release vasoconstrictors that may compromise the microcirculation. Recurrent thrombus formation at the lesion site may result in occlusion of the artery adding to the dynamic nature of the clinical presentation. In conclusion, platelets, the coagulation system, and the endothelium cause a dynamic process of intermittent occlusion, vasospasm and embolization of thrombus material. PMID- 11054909 TI - Optimization of platelet therapy. AB - Percutaneous coronary intervention produces vessel wall injury and activation of platelets that are responsible for producing peri-procedural ischemic complications. The importance of adequate antiplatelet therapy during coronary intervention to reduce platelet mediated ischaemic complications has been recognized for some time. Until recently, adjunctive treatment with aspirin was the only available antiplatelet therapy after coronary intervention that had demonstrated benefit. During the last decade, newer and more potent agents have demonstrated consistent reductions in ischaemic events after intervention and appear to have some enduring effect. Additionally, optimization of antiplatelet therapy with aspirin and the thienopyridines after coronary stenting has been an important advance allowing for the current liberal use of coronary stents. PMID- 11054910 TI - Thrombolytic therapy and percutaneous coronary intervention in unstable angina. PMID- 11054911 TI - The emergence of mechanical thrombectomy; a clot burden reduction approach. AB - Intracoronary thrombi are harbingers for increased procedural complications after percutaneous revascularization techniques. Current approaches to treat coronary thrombus prior to plaque intervention are pharmacologic and mechanical. Whereas the use of coronary thrombolysis prior to or during percutaneous coronary intervention have yielded mixed results, the group of the platelet IIb/IIIa inhibitors have uniformly been shown to decrease the procedural complications and the 30 day rate of major cardiovascular events. Mechanical approaches to managing thrombus include: compression with balloon angioplasty or stenting, removal with atherectomy devices, thromboaspiration with the Possis AngioJet and hydrolyser, and vibration disintegration with the ultrasound thrombolysis device. Recent clinical trials have shown that the Possis AngioJet and the ultrasound thrombolysis device are highly effective and safe for removing coronary thrombi prior to coronary intervention. PMID- 11054912 TI - Catheter-based ultrasound thrombolysis--a new promising thrombus-debulking device for the treatment of intracoronary thrombosis. AB - Angiographic suggestion of intercoronary thrombus is often seen in patients sustaining acute coronary syndromes (ACS). Even in the era of stenting and glycoprotein IIb/IIIa antagonists, the presence of thrombus-rich lesion during percutaneous coronary interventions portends an increased risk of adverse events. It has been hypothesized that reduction of clot-burden prior to PCI may reduce complications and enhance efficacy. Experimental and clinical data have shown that catheter-based ultrasound thrombolysis is capable of inducing an efficacious and safe thrombus-debulking. This article reviews the collective experience with this promising device solution for the treatment of thrombotic lesions in the setting of ACS. PMID- 11054913 TI - Treatment of lesions with thrombus: beyond stents and GP IIb/IIIa inhibitors. AB - In the presence of an intracoronary thrombus, percutaneous coronary intervention (PCI) will frequently lead to complications. Glycoprotein IIb/IIIa blockade as adjunct to PCI is very effective in patients with non-occlusive clots and biochemical evidence of platelet micro-embolization. Thrombotically-occluded vessels still remain a major clinical problem. This provides a rationale for thrombus debulking prior to PCI. A powerful antiplatelet agent used in combination with a thrombus debulking strategy and stenting of the underlying ruptured plaque offers the potential for further enhancement of PCI. Protection against embolization could potentially be optimized with the use of anti embolization devices and covered stents. PMID- 11054914 TI - Brain areas involved in perception of biological motion. AB - These experiments use functional magnetic resonance imaging (fMRI) to reveal neural activity uniquely associated with perception of biological motion. We isolated brain areas activated during the viewing of point-light figures, then compared those areas to regions known to be involved in coherent-motion perception and kinetic-boundary perception. Coherent motion activated a region matching previous reports of human MT/MST complex located on the temporo-parieto occipital junction. Kinetic boundaries activated a region posterior and adjacent to human MT previously identified as the kinetic-occipital (KO) region or the lateral-occipital (LO) complex. The pattern of activation during viewing of biological motion was located within a small region on the ventral bank of the occipital extent of the superior-temporal sulcus (STS). This region is located lateral and anterior to human MT/MST, and anterior to KO. Among our observers, we localized this region more frequently in the right hemisphere than in the left. This was true regardless of whether the point-light figures were presented in the right or left hemifield. A small region in the medial cerebellum was also active when observers viewed biological-motion sequences. Consistent with earlier neuroimaging and single-unit studies, this pattern of results points to the existence of neural mechanisms specialized for analysis of the kinematics defining biological motion. PMID- 11054915 TI - Shifting from the perceptual brain to the logical brain: the neural impact of cognitive inhibition training. AB - What happens in the human brain when the mind has to inhibit a perceptual process in order to activate a logical reasoning process? Here, we use functional imaging to show the networks of brain areas involved in a deductive logic task performed twice by the same subjects, first with a perceptual bias and then with a logical response following bias-inhibition training. The main finding is a striking shift in the cortical anatomy of reasoning from the posterior part of the brain (the ventral and dorsal pathways) to a left-prefrontal network including the middle frontal gyrus, Broca's area, the anterior insula, and the pre-SMA. This result indicates that such brain shifting is an essential element for human access to logical thinking. PMID- 11054916 TI - Performance on indirect measures of race evaluation predicts amygdala activation. AB - We used fMRI to explore the neural substrates involved in the unconscious evaluation of Black and White social groups. Specifically, we focused on the amygdala, a subcortical structure known to play a role in emotional learning and evaluation. In Experiment 1, White American subjects observed faces of unfamiliar Black and White males. The strength of amygdala activation to Black-versus-White faces was correlated with two indirect (unconscious) measures of race evaluation (Implicit Association Test [IAT] and potentiated startle), but not with the direct (conscious) expression of race attitudes. In Experiment 2, these patterns were not obtained when the stimulus faces belonged to familiar and positively regarded Black and White individuals. Together, these results suggest that amygdala and behavioral responses to Black-versus-White faces in White subjects reflect cultural evaluations of social groups modified by individual experience. PMID- 11054917 TI - Visualization and measurement of the cortical surface. AB - Much of the human cortical surface is obscured from view by the complex pattern of folds, making the spatial relationship between different surface locations hard to interpret. Methods for viewing large portions of the brain's surface in a single flattened representation are described. The flattened representation preserves several key spatial relationships between regions on the cortical surface. The principles used in the implementations and evaluations of these implementations using artificial test surfaces are provided. Results of applying the methods to structural magnetic resonance measurements of the human brain are also shown. The implementation details are available in the source code, which is freely available on the Internet. PMID- 11054918 TI - Abnormal functional activation during a simple word repetition task: A PET study of adult dyslexics. AB - Eight dyslexic subjects, impaired on a range of tasks requiring phonological processing, were matched for age and general ability with six control subjects. Participants were scanned using positron emission tomography (PET) during three conditions: repeating real words, repeating pseudowords, and rest. In both groups, speech repetition relative to rest elicited widespread bilateral activation in areas associated with auditory processing of speech; there were no significant differences between words and pseudowords. However, irrespective of word type, the dyslexic group showed less activation than the control group in the right superior temporal and right post-central gyri and also in the left cerebellum. Notably, the right anterior superior temporal cortex (Brodmann's area 22 [BA 22]) was less activated in each of the eight dyslexic subjects, compared to each of the six control subjects. This deficit appears to be specific to auditory repetition as it was not detected in a previous study of reading which used the same sets of stimuli (Brunswick, N., McCrory, E., Price, C., Frith, C.D., & Frith, U. [1999]. Explicit and implicit processing of words and pseudowords by adult developmental dyslexics: A search for Wernicke's Wortschatz? Brain, 122, 1901-1917). This implies that the observed neural manifestation of developmental dyslexia is task-specific (i.e., functional rather than structural). Other studies of normal subjects indicate that attending to the phonetic structure of speech leads to a decrease in right-hemisphere processing. Lower right hemisphere activation in the dyslexic group may therefore indicate less processing of non-phonetic aspects of speech, allowing greater salience to be accorded to phonological aspects of attended speech. PMID- 11054919 TI - Brain activation during mental transformation of size. AB - Visual comparison between different-sized objects with respect to shape can be done by encoding one of the objects as a mental image, transforming the image to the size format of the other object, and then testing for a match (Bundesen, C., & Larsen, A. [1975]. Visual transformation of size. Journal of Experimental Psychology: Human Perception and Performance, 1, 214-220). To identify the brain structures implicated in mental transformation of size, we measured the distribution of regional cerebral blood flow (rCBF) by positron emission tomography (PET) in 12 normal subjects who compared random stimulus patterns with respect to shape regardless of variations in size in a one-back match-to-sample paradigm. Each subject was PET-scanned 12 times during repetitive injections of H(2)(15)O. In one condition (three scans), all stimulus patterns were small. In a second condition (three scans), all stimuli were large. In the third condition (six scans), the stimuli alternated between small and large. Mental transformation of size should occur in the alternating-size condition but not in the fixed-size conditions. As expected, behavioral measures (reaction time [RT], d', beta) were nearly the same for the two fixed-size conditions but mean RT was longer and d' smaller in the alternating-size condition. Changes in rCBF specific to mental transformation of size were estimated by contrasting the alternating size with the fixed-size conditions by use of statistical parametric mapping (SPM96) at a threshold of p <. 05 corrected for multiple comparisons. The detected brain structures implicated in mental transformation of size were primarily located in the dorsal pathways, comprising structures in the occipital, parietal, and temporal transition zone (predominantly in the left hemisphere), posterior parietal cortex (bilaterally), area MT/V5 (left), and vermis (bilaterally). Contrasts between the two fixed-size conditions showed significant effects in only the occipital cortex. PMID- 11054920 TI - The effects of divided attention on encoding- and retrieval-related brain activity: A PET study of younger and older adults. AB - Divided attention (DA) disrupts episodic encoding, but has little effect on episodic retrieval. Furthermore, normal aging is associated with episodic memory impairments, and when young adults are made to encode information under DA conditions, their memory performance is reduced and resembles that of old adults working under full attention (FA) conditions. Together, these results suggest a common neurocognitive mechanism by which aging and DA during encoding disrupt memory performance. In the current study, we used PET to investigate younger and older adults' brain activity during encoding and retrieval under FA and DA conditions. In FA conditions, the old adults showed reduced activity in prefrontal regions that younger adults activated preferentially during encoding or retrieval, as well as increased activity in prefrontal regions young adults did not activate. These results indicate that prefrontal functional specificity of episodic memory is reduced by aging. During encoding, DA reduced memory performance, and reduced brain activity in left-prefrontal and medial-temporal lobe regions for both age groups, indicating that DA during encoding interferes with encoding processes that lead to better memory performance. During retrieval, memory performance and retrieval-related brain activity were relatively immune to DA for both age groups, suggesting that DA during retrieval does not interfere with the brain systems necessary for successful retrieval. Finally, left inferior prefrontal activity was reduced similarly by aging and by DA during encoding, suggesting that the behavioral correspondence between these effects is the result of a reduced ability to engage in elaborate encoding operations. PMID- 11054921 TI - Hemispheric asymmetries for whole-based and part-based face processing in the human fusiform gyrus. AB - Behavioral studies indicate a right hemisphere advantage for processing a face as a whole and a left hemisphere superiority for processing based on face features. The present PET study identifies the anatomical localization of these effects in well-defined regions of the middle fusiform gyri of both hemispheres. The right middle fusiform gyrus, previously described as a face-specific region, was found to be more activated when matching whole faces than face parts whereas this pattern of activity was reversed in the left homologous region. These lateralized differences appeared to be specific to faces since control objects processed either as wholes or parts did not induce any change of activity within these regions. This double dissociation between two modes of face processing brings new evidence regarding the lateralized localization of face individualization mechanisms in the human brain. PMID- 11054922 TI - Asymmetry of neuronal activity during extracellular microelectrode recording from left and right human temporal lobe neocortex during rhyming and line-matching. AB - Recordings of neuronal activity in humans have identified few correlates of the known hemispheric asymmetries of functional lateralization. Here, we examine single-unit activity recorded from both hemispheres during two delayed match-to sample tasks that show strong hemispheric lateralization based on lesion effects; a line-matching (LM) task related to the right hemisphere, and a rhyming (RHY) task related to the left. Nineteen neuronal populations were recorded with extracellular microelectrodes from the left temporal neocortex of 11 awake patients, and 18 from the right in 9 patients during anterior temporal lobectomy for complex partial seizures under local anesthesia. All subjects were left hemisphere dominant for language. Twelve (32%) populations exhibited statistically significant changes in activity at p <.05. Although changes in firing frequency were recorded from both hemispheres during both tasks, the RHY task elicited changes in activity several hundred milliseconds earlier on the left side than on the right. The LM task, on the other hand, induced changes earlier on the right side than on the left. Both hemispheres contained units active during verbal responses regardless of which behavior elicited the response. Our results indicate that cerebral dominance is reflected in earlier neuronal activity in the anterior temporal lobe during tasks lateralized to that hemisphere. PMID- 11054923 TI - Mental maze solving. AB - We sought to determine how a visual maze is mentally solved. Human subjects (N = 13) viewed mazes with orthogonal, unbranched paths; each subject solved 200-600 mazes in any specific experiment below. There were four to six openings at the perimeter of the maze, of which four were labeled: one was the entry point and the remainder were potential exits marked by Arabic numerals. Starting at the entry point, in some mazes the path exited, whereas in others it terminated within the maze. Subjects were required to type the number corresponding to the true exit (if the path exited) or type zero (if the path did not exit). In all cases, the only required hand movement was a key press, and thus the hand never physically traveled through the maze. Response times (RT) were recorded and analyzed using a multiple linear regression model. RT increased as a function of key parameters of the maze, namely the length of the main path, the number of turns in the path, the direct distance from entry to termination, and the presence of an exit. The dependence of RT on the number of turns was present even when the path length was fixed in a separate experiment (N = 10 subjects). In a different experiment, subjects solved large and small mazes (N = 3 subjects). The former was the same as the latter but was scaled up by 1.77 times. Thus both kinds of mazes contained the same number of squares but each square subtended 1.77 degrees of visual angle (DVA) in the large maze, as compared to 1 DVA in the small one. We found that the average RT was practically the same in both cases. A multiple regression analysis revealed that the processing coefficients related to maze distance (i.e., path length and direct distance) were reduced by approximately one-half when solving large mazes, as compared to solving small mazes. This means that the efficiency in processing distance-related information almost doubled for scaled-up mazes. In contrast, the processing coefficients for number of turns and exit status were practically the same in the two cases. Finally, the eye movements of three subjects were recorded during maze solution. They consisted of sequences of saccades and fixations. The number of fixations in a trial increased as a linear function of the path length and number of turns. With respect to the fixations themselves, eyes tended to fixate on the main path and to follow it along its course, such that fixations occurring later in time were positioned at progressively longer distances from the entry point. Furthermore, the time the eyes spent at each fixation point increased as a linear function of the length and number of turns in the path segment between the current and the upcoming fixation points. These findings suggest that the maze segment from the current fixation spot to the next is being processed during the fixation time (FT), and that a significant aspect of this processing relates to the length and turns in that segment. We interpreted these relations to mean that the maze was mentally traversed. We then estimated the distance and endpoint of the path mentally traversed within a specific FT; we also hypothesized that the next portion of the main path would be traversed during the ensuing FT, and so on for the whole path. A prediction of this hypothesis is that the upcoming saccade would land the eyes at or near the locus on the path where the mental traversing ended, so that "the eyes would pick up where the mental traversal left off." In this way, a portion of the path would be traversed during a fixation and successive such portions would be strung together closely along the main path to complete the processing of the whole path. We tested this prediction by analyzing the relations between the path distance of mental traverse and the distance along the path between the current and the next fixation spot. (ABSTRACT TRUNCATED) PMID- 11054924 TI - Impaired processing of complex auditory stimuli in rats with induced cerebrocortical microgyria: An animal model of developmental language disabilities. AB - Individuals with developmental language disabilities, including developmental dyslexia and specific language impairment (SLI), exhibit impairments in processing rapidly presented auditory stimuli. It has been hypothesized that these deficits are associated with concurrent deficits in speech perception and, in turn, impaired language development. Additionally, postmortem analyses of human dyslexic brains have revealed the presence of focal neocortical malformations such as cerebrocortical microgyria. In an initial study bridging these research domains, we found that male rats with induced microgyria were impaired in discriminating rapidly presented auditory stimuli. In order to further assess this anatomical- behavioral association, we designed two experiments using auditory-reflex modification. These studies were intended to assess whether auditory processing deficits in microgyric male rats would be seen in threshold detection of a silent gap in white noise, and in oddball detection of a two-tone stimulus of variable duration. Results showed no differences between sham and microgyric subjects on gap detection, but did show that microgyric subjects were impaired in the discrimination of two-tone stimuli presented in an oddball paradigm. This impairment was evident for stimuli with total duration of 64 msec or less, while both groups were able to discriminate stimuli with duration of 89 msec or greater. The current results further support the relationship between malformations of the cerebral cortex and deficits in rapid auditory processing. They also suggest that the parameters characterizing rapid auditory processing deficits for a specific task may be influenced by stimulus features and/or cognitive demand of that particular task. PMID- 11054925 TI - The role of spatial selective attention in working memory for locations: evidence from event-related potentials. AB - We investigated the hypothesis that the covert focusing of spatial attention mediates the on-line maintenance of location information in spatial working memory. During the delay period of a spatial working-memory task, behaviorally irrelevant probe stimuli were flashed at both memorized and nonmemorized locations. Multichannel recordings of event-related potentials (ERPs) were used to assess visual processing of the probes at the different locations. Consistent with the hypothesis of attention-based rehearsal, early ERP components were enlarged in response to probes that appeared at memorized locations. These visual modulations were similar in latency and topography to those observed after explicit manipulations of spatial selective attention in a parallel experimental condition that employed an identical stimulus display. PMID- 11054926 TI - Fine discrimination of faces can be performed rapidly. AB - Here we measure the smallest change in a face that can be discriminated. A morphing algorithm mixed two faces in variable proportions to create a series of synthetic faces that each differed by a tiny amount. By selecting from this series, a test face could be chosen so as to reach a just noticeable difference from a sample face. Face-discrimination thresholds were about 7% of the average difference between two faces, as quantified by coefficients of a principal components decom position. This threshold remained constant as the duration of the test face was reduced from 1,000 to 100 msec, and rose quickly for shorter stimulus durations. The behavioral evidence presented here indicates that complex visual processing can be completed within the first 100 msec of the signal, suggesting involvement of feedforward neural mechanisms, and placing constraints on possible computational algorithms employed within the ventral visual pathways. PMID- 11054927 TI - Grasping after a delay shifts size-scaling from absolute to relative metrics. AB - We carried out three experiments designed to compare the effects of relative and absolute size on manual prehension and manual estimates of perceived size. In each experiment, right-handed subjects were presented with two different-sized 3 D objects in a virtual display and were instructed to pick up or estimate the size of one of them. In Experiment 1, subjects were requested to pick up the smaller one of two virtual objects under one condition and the larger one under the other condition. In fact, the target object was identical on all trials; it was simply paired with a smaller object on some trials and a larger object on others. To provide veridical haptic feedback, a real object was positioned beneath a mirror at the same location as the virtual target object. In Experiment 2, one of the virtual objects was marked with a red dot on its top surface. From trial to trial, the marked object was paired with a larger, smaller, or same sized object. Subjects were instructed to always pick up the marked object on each trial. In both Experiment 1 and 2, half the subjects were tested in delayed grasping with a 5-sec delay between viewing the objects and initiating the grasp, and half in real-time grasping without a delay. Using the same display of virtual objects as in Experiment 2, subjects in Experiment 3 were requested to estimate the size of the marked object using their index finger and thumb (i.e., they showed us how big the object looked to them). After estimating the target object's size, they picked it up. All subjects gave their estimates either immediately or after a delay. Recording of hand movements revealed that when subjects in Experiments 1 and 2 picked up the target object in real time, their grip aperture in flight was not significantly affected whether the object was accompanied by a larger object or a smaller one. When subjects picked up the target object after a delay, however, their grip aperture in flight was larger when the target object was accompanied by a smaller object than when it was accompanied by a larger object. A similar size-contrast effect was also observed in Experiment 3 in which subjects gave manual estimates of the perceived size of the target object. This perceptual effect was observed both when the estimates were given immediately and when they were given after a 5-sec delay. These results suggest that normal (real-time) visuomotor control relies on absolute metrics, whereas delayed grasping utilizes the same relative metrics used by conscious perception. PMID- 11054928 TI - Electrophysiological correlates of conscious vision: evidence from unilateral extinction. AB - To study the electrophysiological correlates of conscious vision, we recorded event-related potentials (ERPs) in a patient with partial unilateral visual extinction as a result of right-hemisphere damage. When, following bilateral presentations, contralesional stimuli were not perceived, there was an absence of the early attention-sensitive P1 (80-120 msec) and N1 (140-180 msec) components of the ERP response. In contrast, following unilateral presentations, or in those bilateral presentations in which contralesional stimuli were perceived (about 60%), these ERP components were present. These results provide novel evidence that extinction involves the stage of early focusing of attention and that the P1 and N1 components of visual ERPs are reliable physiological correlates of conscious vision. PMID- 11054929 TI - Topographic and temporal indices of vowel spectral envelope extraction in the human auditory cortex. AB - The auditory-evoked neuromagnetic field elicited by single vowel formants and two formant vowels was recorded under active listening conditions using a 37-channel magnetometer. There were three single formants with formant frequencies of 200, 400, and 800 Hz, another single formant with a formant frequency of 2600 Hz, and three vowels that were constructed by linear superimposition of the high- onto one of the low-frequency formants. P50 m and N100 m latency values were inversely correlated with the formant frequency of single formants. A strong effect of formant frequency on source location was obtained along the postero-anterior axis, which is orthogonal to the well-established latero-medial tonotopic gradient. Regardless of whether single formants or first formants of vowels were considered, N100 m sources were more anterior and sustained field sources were more posterior for higher-frequency than for lower-frequency formants. The velocity of the apparent posterior-to-anterior movement across cortical surface of N100 m sources first reported by Rogers et al. [Rogers, R. L., Papanicolaou, A. C., Baumann, S. B., Saydjari, C., & Eisenberg, H. M. (1990). Neuromagnetic evidence of a dynamic excitation pattern generating the N100 auditory response. Electroencephalography and Clinical Neurophysiology,77, 237-240] decreased as a function of latency. The amount of deceleration was positively correlated with formant frequency. Responses to the vowels were superadditive, indicating that the processes elicited by the constituents of composite stimuli interact at one or more stages of the afferent auditory pathway. Such interaction may account for the absence of a lateral-to-medial tonotopic mapping of first formant frequency. The source topography found may reflect activity in auditory fields adjacent to AI with the strength of the contribution varying with formant frequency. Alternatively, it may reflect sharpness-of-tuning and inhibitory response-area asymmetry gradients along isofrequency stripes within AI. Either alternative may be interpreted in terms of a spectral blurring mechanism that abstracts spectral envelope information from the details of spectral composition, an important step towards the formation of invariant phonetic percepts. PMID- 11054931 TI - Re: Color vision and the four-color-map problem, by Purves et al. (JOCN 12:2, 233 237, 2000). PMID- 11054930 TI - The strategic control of gaze direction in the Tower-of-London task. AB - In this paper, we describe a novel approach to the study of problem solving involving the detailed analysis of natural scanning eye movements during the "one touch" Tower-of-London (TOL) task. We showed subjects a series of pictures depicting two arrangements of colored balls in pockets within the upper and lower halves of a computer display. The task was to plan (but not to execute) the shortest movement sequence required to rearrange the balls in one half of the display (the Workspace) to match the arrangement in the opposite half (the Goalspace) and indicate the minimum number of moves required for problem solution. We report that subjects are more likely to look towards the Goalspace in the initial period after picture presentation, but bias gaze towards the Workspace during the middle of trials. Towards the end of a trial, subjects are once again more likely to fixate the Goalspace. This pattern is found regardless of whether the subjects solve problems by rearranging the balls in the lower or upper visual fields, demonstrating that this strategy correlates with discrete phases in problem solving. A second experiment showed that efficient planners direct their gaze selectively towards the problem critical balls in the Workspace. In contrast, individuals who make errors spend more time looking at irrelevant items and are strongly influenced by the movement strategy needed to solve the preceding problem. We conclude that efficient solution of the TOL requires the capacity to generate and flexibly shift between control sets, including those underlying ocular scanning. The role of working memory and the prefrontal cerebral cortex in the task are discussed. PMID- 11054932 TI - Reply PMID- 11054933 TI - Erratum PMID- 11054934 TI - XIIIth International AIDS Conference: a continuing update. PMID- 11054935 TI - Herpes zoster in an HIV-negative man on ritonavir. PMID- 11054936 TI - Persistent numbness and burning pain in a man with HIV disease. PMID- 11054937 TI - Physiology, immunology, and disease transmission in human breast milk. AB - Breast feeding is an important mode of mother-to-child transmission of HIV. Interventions to decrease the number of infants becoming infected are particularly required for women in less developed countries where breast feeding is essential for infant survival. This review discusses the physiology and immunology of breast feeding and how maternal health interventions in the postpartum period may help decrease mother-to-child HIV transmission. PMID- 11054938 TI - Acute rhabdomyolysis and renal failure in HIV-infected patients: risk factors, presentation, and pathophysiology. AB - Rhabdomyolysis is a common cause of acute renal failure and may be related to a variety of predisposing factors. This entity has been increasingly recognized in HIV-infected individuals and is an important cause of morbidity and mortality. We present a series of seven HIV-positive patients admitted with rhabdomyolysis over a 5-year period; three developed acute renal failure. Infections and substance abuse were the most common risk factors identified; an average of three predisposing factors was present in each case. All patients showed resolution of creatinine phosphokinase (CPK) elevation and serum creatinine returned to the normal range in the three patients who developed renal insufficiency; however, all patients required prolonged hospitalization and one patient died of sepsis. The pathophysiological mechanisms of muscle injury in our patients are reviewed and their bearing on prognosis discussed. It is concluded that clinicians should have a high index of suspicion for the development of rhabdomyolysis in HIV infected patients with a combination of noncompliance with medical therapy and/or substance abuse and acute infection. With comprehensive supportive care, the prognosis of acute rhabdomyolysis in this population may be reasonably good. PMID- 11054939 TI - Medication usage and dental caries outcome-related variables in HIV/AIDS patients. AB - The purpose of this study was to access current medication usage by HIV/AIDS patients and its effects on dental caries and on unstimulated salivary flow rates. Thirty females and 127 males (mean age = 39.6 +/- 7.4 years), of whom 46% were White/Non-Hispanic, 39% African-American, and 15% Hispanic, were examined and interviewed at the Bering Dental Clinic, Houston, Texas. The mean time in years after seroconversion was 5.4 +/- 4.1. Calibrated examiners performed dental caries examination (DMFS) with dental explorers and bitewing radiographs. Interviews were carried out with pretested questionnaires, and medication usage was assessed by illustrative examples of HIV/AIDS medications. Salivary flow rates were determined gravimetrically (mL/min). Bivariate analysis and analysis of variance (ANOVA) were used to analyze the data. Because there were no race or gender effects on dental caries outcome variables or salivary flow rates, separate logistic regression models for medication usage were generated, which were adjusted for age and CD4+ cell counts. Patients who, currently, were receiving antiretroviral therapy (ART) had a lower occurrence of dental caries than patients not taking these medications. An unexpected finding in the lower caries rate group was a decrease in salivary flow rates, which was a probable oral side effect of ART. It appears from this cross-sectional study that systemic medication for the management of HIV disease has no significant detrimental effect on the dentition. PMID- 11054940 TI - The multidimensional context of HIV/AIDS patient care. AB - This article argues that enhancing efficacious social networks or developing new adaptive social ties for HIV-infected individuals without adequate social resources may enhance their adaptive response to the disease and its treatment. It further suggests that the success of an adaptive response to the disease may to some extent depend on the relationship between the patient and the healthcare provider(s). PMID- 11054941 TI - Who receives Ryan White CARE Act services? A demographic comparison of CARE act clients and the general AIDS population. AB - This study examines the extent to which health and social service providers funded by the Ryan White Comprehensive AIDS Resources Emergency (CARE) Act serve women, minorities, and other vulnerable populations emphasized by the legislation. Demographic characteristics of AIDS-diagnosed clients served by CARE Act-funded providers in four metropolitan areas and two states are compared with Centers for Disease Control and Prevention estimates of AIDS prevalence. Clients of CARE Act-funded providers tend to reflect the demographics of local HIV/AIDS epidemics. Where differences exist, CARE Act clients are more likely to be women and minorities and less likely to be injecting drug users. CARE Act-funded providers are effectively reaching most medically underserved populations. PMID- 11054942 TI - PIs may cause sexual dysfunction. PMID- 11054943 TI - Teens more reluctant to have sex. PMID- 11054944 TI - Effect of statin therapy on restenosis after coronary stent implantation. PMID- 11054945 TI - Phentermine's effect on serotonin in plasma? PMID- 11054946 TI - Dispersion in electrocardiography has become epidemic. PMID- 11054947 TI - The business of medicine. Ballot initiatives offer physicians a new response to managed care cost-cutting. PMID- 11054948 TI - Lesion in an old scar. Biopsy of eroded papule is required to rule out potentially serious diagnoses. PMID- 11054949 TI - High risk on the highway. How to identify and treat the impaired older driver. AB - Among older adults who drive, the rate of those involved in fatal crashes rises after age 70. Problem driving in older adults involves visual, cognitive, and motor skills, which may decline with aging and chronic disease. Physicians and other health care providers may not be prepared to evaluate and advise the older patient on the emotion-loaded topic of driving ability. Although it is difficult to identify prospectively a high-risk driver, a targeted history and physical exam are useful clinical tools. Two models are proposed to guide the health care provider and patient through the process of limiting or ending the impaired older driver's time behind the wheel. PMID- 11054950 TI - Geriatric pain. Factors that limit pain relief and increase complications. AB - Persons age 65 and older are more likely than younger adults to experience chronic pain but less likely to obtain pain relief. Achieving adequate pain management for the older patient is complicated by comorbid diseases, increased risk of adverse drug reactions, and physician factors such as inadequate training in pain medicine and a reluctance to prescribe opioid medications. Nociception appears not to change with age or with the development of dementia, although a person's perception of pain and willingness to report it may change. Control of depression and anxiety greatly facilitates pain management. As a patient's number of medications increases, so does the risk of adverse reactions; therefore, care is required when adding any new medication to the drug regimen. PMID- 11054951 TI - Late-life dementia. Review of the APA guidelines for patient management. AB - Management of dementia in older patients requires an individualized and multimodal approach that involves use of psychiatric, psychotherapeutic, psychosocial, and somatic tools and treatments, in addition to patient and family education. The progressive nature of dementia and the invariable presence of comorbidity complicates the management task, although symptoms characteristic of dementia's phases can provide helpful clinical clues to guide evolving care. In 1997, the American Psychiatric Association published the "Practice guideline for the treatment of patients with Alzheimer's disease and other dementias of late life." To date, this is the most comprehensive clinical guideline available to physicians caring for patients with Alzheimer's disease and other related dementias. PMID- 11054952 TI - Older Americans 2000. New data system that tracks health and well-being finds successes and disparities. PMID- 11054953 TI - Why physicians need to know more about aging. PMID- 11054954 TI - Rapid evolution of acute myocardial infarction. PMID- 11054955 TI - Case in point. Cerebral toxoplasmosis. PMID- 11054956 TI - A woman with pansinusitis and cranial nerve palsies. PMID- 11054957 TI - Smoking cessation: when to intervene. PMID- 11054958 TI - Screening in the elderly: principles and practice. PMID- 11054959 TI - Planning and providing care at the end of life. PMID- 11054960 TI - Xenotransplantation: pausing to reflect. PMID- 11054961 TI - Normal anatomy of the neck with CT and MR imaging correlation. AB - Knowledge of normal anatomy is the cornerstone for understanding pathologic processes. Often, the most difficult task for a radiologist is differentiating normal structures from pathology. There are many ways to organize a discussion of the normal anatomy in the neck. This article is organized by organ system, with an additional discussion of triangles, fascia, and spaces. Important normal variants are also discussed. PMID- 11054962 TI - CT and MR imaging evaluation of neck infections with clinical correlations. AB - Infection of the neck is a common clinical problem in all age groups, especially children and young adults. The clinical symptoms and signs are often suggestive of the diagnosis. Imaging studies including CT and MR imaging are frequently required to confirm the diagnosis but more importantly to localize the infectious process and search for and delineate an abscess cavity. Ultrasound has also been used in the evaluation of superficial neck infections, especially to determine fluid accumulation. Conventional films consisting of an anteroposterior and lateral view were the examination before the introduction of CT in 1972. Conventional films can still be used for a preliminary survey, especially of the retropharyngeal space when there is a question of a retropharyngeal phlegmon or abscess. PMID- 11054963 TI - Imaging of granulomatous lesions of the neck in children. AB - Cervical lymphadenopathy is the most common presentation of granulomatous inflammation of the neck in children and is usually caused by NTM infection. Although certain granulomatous infections have characteristic imaging features, there is considerable overlap in the imaging appearance of the various disorders. The diagnosis is usually based on a combination of clinical features, histopathologic examination, serologic tests, and culture results. PMID- 11054964 TI - Lymph node pathology. Benign proliferative, lymphoma, and metastatic disease. AB - The evaluation of cervical lymph nodes is one of the main indications for performing CT and MR imaging of the neck. Imaging may be done for evaluation of an unknown neck mass, but more commonly the neck is imaged to evaluate potential metastasis from a known mucosal malignancy. CT and MR imaging characteristics of both malignant and nonmalignant nodal diseases are reviewed and the differential diagnosis of nodal pathologies for specific imaging findings are discussed. A recently proposed imaging-based nodal classification for metastatic nodal diseases from head and neck cancer is also described. PMID- 11054965 TI - The indications of FDG-PET in neck oncology. AB - FDG-PET imaging in neck oncology has a definite clinical impact in the post therapy setting, assisting in the management of thyroid cancers and SCC of the neck. Quantitation of FDG uptake in suspicious areas may be helpful but should be regarded cautiously. Overall, wider incorporation of FDG imaging in clinical routine depends also on cost availability issues of FDG and of imaging devices. Dual-coincidence scanners for FDG imaging are much cheaper than dedicated PET scanners and are installed in growing numbers in many centers. These devices have inferior sensitivity; however, series published with these scanners produce encouraging results. Easier and more acceptable clinical application will also be facilitated by the systematic use of coregistration with anatomic images. Both prerequisites might be fulfilled by the emergence on the market of a gamma camera mounted anatomic X-ray tomograph, which in addition to dual-coincidence scintigraphic imaging provides radiographic images of comparable quality to third generation CT systems. This type of hybrid gamma camera-CT scanner has great potential in a region of complex anatomy, such as the head and neck. PMID- 11054966 TI - Imaging of cystic lesions. AB - Cystic neck masses are varied in their histology and embryogenesis. Because neural, vascular, and lipomatous lesions may all appear cystic, a multimodality imaging approach can help identify these potential mimics. Developmental neck cysts include thyroglossal duct, thymic, and branchial cleft cysts, and teratomatous lesions or lymphangiomas. Although laryngoceles are acquired lesions, congenital anomalies (e.g., abnormally long saccules) may play a role in their formation. Lesion location is at least as important a determinant as morphology in formulating the differential diagnosis of a cystic neck mass. Midline cystic lesions are most commonly thyroglossal duct cysts, although dermoid tumors are also frequently midline. Squamous cell carcinoma metastatic to anterior triangle lymph nodes (Fig. 17), and cystic, necrotic schwanommas, can mimic the typical appearance of an infected second branchial cleft cyst. Posterior triangle lymphadenopathy and lipomatous lesions may resemble cystic hygromas. Cystic-appearing masses in the carotid space include neurogenic tumors, vascular thromboses, and carotid chain lymphadenopathy. Neural-based lesions typically occur posterolateral to the carotid artery. Necrotic lymphadenopathy may be suggested by lesion multiplicity, or by the presence of ancillary features, such as systemic symptoms, or the existence of a primary tumor. It must be emphasized that the primary role of the radiologist in head and neck imaging is to help stage disease and guide surgery. Despite clinical and radiographic analysis, the diagnosis of many lesions ultimately depends on image-guided or excisional biopsy. PMID- 11054967 TI - Ultrasound of the neck. AB - Sonography, when performed by an experienced examiner, can be used for evaluation of many pathologies in the head and neck area. Some benign neck lesions, such as cysts, lipomas, carotid body tumors, and hyperplastic lymph nodes, have typical sonomorphology. Sonography has an accuracy rate of about 90% in cervical lymph node staging and can delineate subclinical lymph node recurrences. It is the method of choice for evaluation of tumor infiltrations of the wall of the great vessels. Salivary gland tumors in the superficial lobe can be delineated completely by sonography. Salivary stones can be detected and localized. Carcinoma of the tongue and floor of the mouth with T1 and T2 staging can be assessed by US. The use and contribution of color Doppler sonography for the assessment of pathologic entities in the neck is a method under clinical investigation. US-guided fine-needle aspiration biopsy of lymph nodes and tumors of the salivary glands is easy to perform and is characterized by high sensitivity and specificity. To perform US examinations of the head and neck area of the highest quality the examiner should be familiar with the anatomy of the head and neck, be informed about the clinical problem, and have experience in the interpretation of abnormal US findings. US of the head and neck area is one of the most difficult sonographic examinations and should be performed by an experienced physician. PMID- 11054968 TI - Imaging of salivary gland pathology. AB - Calcifications in the salivary glands may occur in calculi, chronic infection or inflammation, phleboliths, lymph nodes, chronic hematoma, amyloidosis, and tumors including pleomorphic adenomas, schwannomas, and mucoepidermoid cancers. This article discusses the different types of salivary gland conditions and the best methods of imaging for diagnosis. PMID- 11054969 TI - Glomus faciale, glomus jugulare, glomus tympanicum, glomus vagale, carotid body tumors, and simulating lesions. Role of MR imaging. AB - In summary, MR imaging characteristics of a case of paraganglioma of the facial nerve are reported. The relationship of paragangliomas and the chromaffin system have been discussed. There are many reports of cases of synchronous paragangliomas and pheochromocytomas. These reports, along with simultaneous involvement in familial MEN syndromes, and the common embrylogic origin (neural crest) and similar histopathologic relationships between paragangliomas and pheochromocytoma, all support the fact that they are part of the chromaffin system. PMID- 11054970 TI - Neurogenic tumors of the neck. AB - Neurogenic tumors of the neck occur in children and adults. Important parameters to aid in the differential diagnosis are age at presentation, location, and a history of NF or multiple endocrine neoplasia. Schwannoma is the most common solitary neurogenic tumor in the neck and is usually seen in patients between 20 and 50 years of age. The plexiform neurofibroma and multiple localized neurofibromas are characteristic of NF1. MPNSTs are uncommon aggressive lesions that can arise de novo in patients with NF (10% to 30%) and postirradiation. Neuroblastic tumors consist of neuroblastoma, ganglioneuroblastoma, and ganglioneuroma. These tumors typically arise in the chest and abdomen but occasionally present as a primary neck mass. A neck mass with a histologic diagnosis of neuroblastoma is, however, more commonly metastatic from an abdominal neuroblastoma. PMID- 11054971 TI - Imaging of benign and malignant soft tissue tumors of the neck. AB - Soft tissue tumors of the neck are a heterogeneous group of neoplasms arising from adipose, muscular, and fibrous tissue. With the exception of lymphomas, they account for only a small fraction of neck masses. Nevertheless, accurate diagnosis is important since the behavior of these neoplasms differs markedly from each other and from other head and neck masses. Noninvasive imaging, usually with CT and MR imaging, plays an important role in diagnostic evaluation and treatment planning for these tumors. In some cases, imaging features may be suggestive of a single entity. In most cases, imaging is needed to assess the location and extent of the tumor prior to biopsy or excision. This article discusses imaging techniques used for such assessment, the imaging features that help to separate these neoplasms from other head and neck tumors, and the behavior and imaging features of each of the more common benign and malignant soft tissue tumors that occur in the neck. PMID- 11054972 TI - The thyroid and parathyroid glands. CT and MR imaging and correlation with pathology and clinical findings. AB - Thyroid imaging approach is based on the preliminary clinical evaluation. Lesions that are smaller than 2 cm should be assessed with US, which is capable of discriminating masses as small as 2 mm and distinguishing solid from cystic nodules. US-guided FNAB provides tissue for cytologic examination of thyroid nodules. CT and MR imaging are indicated for larger tumors (greater than 3 cm diameter) that extend outside the gland to adjoining structures, including the mediastinum, and retropharyngeal region. Metastatic lymph nodes in the neck and invasion of the aerodigestive tract are also in the realm of CT and MR imaging. Thyroid nodules are categorized on scintigraphy as hot or cold nodules. Hot nodules are rarely malignant, whereas cold nodules have an incidence of 10% to 20% of malignancy. Calcifications (amorphous, globular, nodular, and linear) occur in adenomas and carcinomas and have no differential diagnostic features except for psammomatous calcifications, which are a pathognomonic finding in papillary carcinomas and a small percentage of medullary carcinomas. Papillary carcinoma is the most common malignant tumor (80%) followed by follicular (20% to 25%); medullary (5%); undifferentiated; anaplastic carcinomas (< 5%); lymphoma (5%); and metastases. Lymph node metastases are common in papillary carcinoma, 50% at presentation, and less common in follicular carcinomas. The metastatic nodes in papillary carcinoma may enhance markedly (hypervascular); show increased signal intensity on T1-weighted images (increased thyroglobulin content or hemorrhage); and reveal punctate calcifications. Localized invasion of the larynx, trachea, and esophagus occurs predominantly in papillary and follicular carcinomas; the incidence is less than 5%. Ectopic thyroid tissue may be encountered in the tongue (foramen cecum); along the midline between posterior tongue and isthmus of thyroid gland; lateral neck; mediastinum; and oral cavity. Goiter and malignant tumors, notably papillary carcinoma, may develop in ectopic thyroid tissue. Carcinomas may also arise in thyroglossal duct cysts, which develop from duct remnants between the foramen cecum and thyroid isthmus. Infectious disease of the thyroid gland is not common and the CT and MR imaging findings are similar as described under neck infection. Other types of inflammatory disorders including Hashimoto's thyroiditis, granulomatous thyroiditis, and Riedel's struma display no specific imaging features. Imaging studies may, however, be indicated to confirm a suspected clinical diagnosis and assess compromise of the airway (Riedel's struma). HPT is a clinical diagnosis in which hypercalcemia is the most important finding. Parathyroid hyperplasia, adenoma, and carcinoma represent underlying lesions. To relieve the patient's symptoms surgical extirpation is indicated. The surgical success rate without imaging is 95%. The indications for imaging studies vary but it is generally agreed that reoperation after a previous failed surgical attempt and suspicion of an ectopic parathyroid adenoma should be investigated by imaging. These consist of US, nuclear medicine studies, CT and MR imaging. US and technetium sestamibi scanning have the highest accuracy rate for localizing an adenomatous gland at and near the thyroid gland. Ectopic adenomas, particularly if they are located in the mediastinum, are preferrably investigated with CT and MR imaging with gadolinium and fat suppression. Carcinomas and parathyroid cysts are optimally evaluated by CT and MR imaging. On MR imaging adenomas are low in signal intensity on T1-weighted images, high in signal intensity on T2-weighted images, and enhance post introduction of gadolinium. PMID- 11054973 TI - Sonography of the thyroid and parathyroid glands. AB - Sonography is the first line modality for assessment of thyroid and parathyroid pathologies. Sonographic and color Doppler patterns of diffuse and focal pathologies of the thyroid are presented in this article. The accuracy of sonography in the localization of enlarged parathyroid glands is also discussed. The limitations of sonography in specifying focal thyroid diseases and the problems in localizing ectopic parathyroid adenoma are addressed. PMID- 11054974 TI - Computerized system for outcomes-based antiemetic therapy in children. AB - OBJECTIVE: To introduce a computerized data collection system used for an outcomes-based approach to antiemetic therapy in children, and to present data collected with this system in support of a new antiemetic dosing regimen. METHODS: A validated nausea/vomiting survey was used to collect data on nausea severity (NSEV), vomiting severity (VSEV), daily activity interference (DAI), and the number of vomiting episodes. NSEV, VSEV, and DAI were rated as 0 = none to 3 = severe. All children and/or their parents were surveyed following the completion of each highly emetogenic chemotherapy regimen. This survey was computerized and transferred to a handheld data entry unit. Time and motion studies were performed to compare the time required to collect nausea/vomiting data and transfer the data to a computerized database with the hand-held system versus traditional paper (manual) surveys. The hand-held technology was used to collect survey data for children receiving a new antiemetic regimen (daily ondansetron and dexamethasone [OD]), which was then compared with data obtained with a previously employed regimen (thrice-daily ondansetron and daily methylprednisolone [OM]). Statistical analysis and a cost-effectiveness analysis (CEA) were performed to compare the two antiemetic regimens. RESULTS: The mean time required for total data entry with the manual system was 5.2 minutes per survey versus 2.4 minutes with the hand-held technology (p = 0.0026). A total of 376 nausea/vomiting surveys in 78 children receiving the OM antiemetic regimen were compared with 153 surveys in 38 children treated with the OD regimen. The mean survey scores were as follows: NSEV (1.2 vs. 0.8), VSEV (1.0 vs. 0.7), DAI (1.0 vs. 0.7), and number of vomiting episodes (4.3 vs. 2.1) for OM and OD, respectively; all were significantly lower with the OD regimen (p < 0.05). The percentage of patients with complete control of nausea and vomiting (19.2% vs. 39.2%) and good control (55.6% vs. 65.4%) were significantly greater with the OD regimen (p < 0.05). The CEA revealed that the OD resulted in a reduction of approximately $31 per patient for good protection and a $258 reduction for complete protection from nausea and vomiting. CONCLUSIONS: A computerized outcomes-based system aided by handheld technology allowed for more prompt and efficient collection of nausea/vomiting data. The OD antiemetic regimen was shown to be a more cost-effective alternative for children receiving severely emetogenic chemotherapy. PMID- 11054975 TI - Adequacy of a new chlorhexidine-bearing polyurethane central venous catheter for administration of 82 selected parenteral drugs. AB - OBJECTIVE: To screen 82 commonly used parenteral medications for compatibility with a new chlorhexidine-bearing central venous catheter, the ARROWg+ard Blue Plus. Evaluations were performed for completeness of drug delivery and impact, if any, of the drugs on the amount of chlorhexidine removed from the internal lumens. DESIGN: Drug solutions were prepared in dextrose 5% injection or NaCl 0.9% at common concentrations. Three 10-mL aliquots of each drug solution were delivered over 10 minutes, one aliquot through each lumen of the triple-lumen catheter. The initial drug concentrations of the admixtures and the effluent samples were analyzed by HPLC for chlorhexidine content and for the amount of drug delivered relative to its initial concentration. RESULTS: The delivery of the infusion solutions alone through sample catheters resulted in no more than trace amounts of chlorhexidine in the solution. Background amounts ranged from < 2.5 to 6.1 micrograms/mL in the first 10 mL of solution. Administration of none of the drugs tested resulted in a substantial increase in chlorhexidine delivery. Furthermore, delivery of most of the drugs was at least 95% and usually was in excess of 97% of the initial concentration. Concentrations of five drugs, amikacin sulfate, cefoperazone sodium, cefotaxime sodium, cefepime HCl, and netilmicin sulfate were somewhat lower than the initial concentration (range 91 95%), but were still considered acceptable. CONCLUSIONS: The ARROWg+ard Blue Plus central venous catheter can be recommended for use with all of the 82 parenteral drugs tested. PMID- 11054976 TI - Congruence of three self-report measures of medication adherence among HIV patients. AB - OBJECTIVE: To evaluate the accordance between three measures of self-reported medication adherence. METHODS: A survey was administered to HIV patients. The three adherence measures included a four-item Morisky-type scale (Measure1) and two measures defining adherence as the percentage of doses taken as prescribed during the past two days (Measure2) or past two weeks (Measure3). RESULTS: For Measure1, 29.2% of the patients were categorized as high adherence and 61.5% as medium adherence. For Measure2 and Measure3, the mean scores were 93.6% and 96.5%, respectively. Using 90% as cutoff values, 78.5% and 95.4% of the patients were classified as adherent by Measure2 and Measure3, respectively (kappa = 0.30; p = 0.001). When 80% was used, 90.8% of the patients were classified as adherent for Measure2 and 96.9% for Measure3 (kappa = 0.48; p < 0.001). When using 90% and 80% as cutoff values to categorize Measure2 and Measure3 as three levels, there was no agreement between Measure1 and the other two measures. The accordance of Measure2 and Measure3 was significant albeit not high (kappa = 0.31; p < 0.001). CONCLUSIONS: The accordance between a Morisky-type adherence scale and measures of missed doses is unsatisfactory. However, "missed-dose" measures using two-day or two-week time periods yield fairly similar results. Researchers should be cautious when comparing adherence rates between studies that use different methods for assessing adherence. PMID- 11054977 TI - Publication rates of abstracts from two pharmacy meetings. AB - OBJECTIVE: To determine the rate of publication of abstracts presented at the 1994 American Society of Health System Pharmacists (ASHP) Mid-year Clinical Meeting and the 1994 American College of Clinical Pharmacists (ACCP) Annual Meeting. METHODS: Abstracts presented at the 1994 ASHP Midyear Clinical Meeting and the 1994 ACCP Annual Meeting were evaluated for subsequent publication as full articles in journals indexed in MEDLINE, International Pharmaceutical Abstracts, and Current Contents. RESULTS: Five hundred one abstracts presented at the 1994 ASHP Mid-year Clinical Meeting were evaluated; 55 (11%) of these had been published. Two hundred fifteen abstracts presented at the 1994 ACCP Annual Meeting were evaluated; 71 (33%) of these had been published. CONCLUSIONS: The publication rates for abstracts presented at ASHP and ACCP meetings were found to be lower than many of those for other medical groups. The presentation of research abstracts at professional meetings is an integral part of the exchange of scientific information; however, many of the presented abstracts are not subsequently published as full research reports. The failure to publish the results of the studies may limit the ability of a reader to judge the validity, reliability, and generalizability of the research. This could affect the use of the findings in clinical practice and in supporting or refuting other research findings. PMID- 11054978 TI - Olanzapine-induced acute pancreatitis. AB - OBJECTIVE: To report the first published case of olanzapine-induced acute pancreatitis. CASE SUMMARY: A 72-year-old white woman was admitted to the intensive care unit (ICU) with acute hemorrhagic pancreatitis and unintentional verapamil overdose. The patient did not consume alcohol and had undergone a cholecystectomy in the past; other medical causes of pancreatitis had been ruled out. She was taking several medications chronically, but olanzapine was started six days prior to the onset of acute abdominal symptoms. According to the Naranjo probability scale, olanzapine was considered the probable cause of acute pancreatitis in this patient. Following a 12-day stay in the ICU, the patient was transferred to the ward where she died a few days later of unrelenting peritonitis secondary to acute pancreatitis. DISCUSSION: A literature search (1966-July 2000) and contact with the manufacturer failed to detect any published reports of acute pancreatitis associated with olanzapine. The contribution of concomitant medications taken prior to ICU admission in initiating or worsening the pancreatitis was deemed unlikely. More common causes of acute pancreatitis, such as ethanol consumption and gallstones, were also ruled out in this patient. Therefore, olanzapine was rated as a probable cause for acute pancreatitis in our patient. The mechanism of this adverse reaction is unknown. CONCLUSIONS: This is believed to be the first published report suspecting olanzapine, an atypical antipsychotic agent, to have caused acute hemorrhagic pancreatitis in a patient, leading to admission to the ICU and, eventually, death secondary to unrelenting peritonitis. PMID- 11054979 TI - Accidental over-anticoagulation: substitution error by a foreign pharmacy. AB - OBJECTIVE: To describe an episode of inadvertent and excessive anticoagulation caused by mistaken substitution of medication by a pharmacy outside the US. CASE SUMMARY: A 57-year-old white woman was found to have profound prolongation of her prothrombin time (56.9 sec) and international normalized ratio (22.18), with other coagulation parameters relatively normal. She had no prior history of bleeding diatheses and was not taking any prescribed anticoagulants. Her coagulopathy rapidly corrected with the administration of fresh frozen plasma and vitamin K. After her medications were visually inspected, it was discovered that she had purchased her prescription medications from a pharmacy in Mexico and that she inadvertently had been taking a preparation of warfarin (proprietary name in Mexico, "Romesa") instead of the prescribed ramipril for her hypertension (proprietary name in Mexico, "Ramace"). After removal of the incorrect medication, she experienced no further prolongation of her coagulation parameters. DISCUSSION: Medication errors contribute significantly to adverse events for patients. The frequency of different types of medication errors is reviewed, and problems specific to the use of warfarin are detailed. Circumstances that might lead to a patient seeking prescription medication outside of the US are also discussed. CONCLUSIONS: The acquisition of prescription medications from pharmacies outside of the US can have adverse consequences, especially if the foreign name of the medication is different from its American name, while sounding similar to other medications that also might be dispensed in foreign pharmacies. PMID- 11054980 TI - Fulminant peripheral neuropathy with severe quadriparesis associated with vincristine therapy. AB - OBJECTIVE: To report a case of fulminant neuropathy with severe quadriparesis associated with vincristine chemotherapy. CASE SUMMARY: A 48-year-old white man with acute lymphoblastic leukemia was started on an induction chemotherapeutic regimen that included intravenous vincristine. He received a total of 6 mg of vincristine over two weeks during induction chemotherapy. Over the next two weeks, he developed a fulminant peripheral neuropathy with severe quadriparesis. DISCUSSION: Although commonly associated with peripheral neuropathy, vincristine neurotoxicity only rarely involves instances of fulminant peripheral neuropathy with severe quadriparesis. Guillain-Barre syndrome is also associated with leukemia and may present as a fulminant peripheral neuropathy with severe quadriparesis. CONCLUSIONS: Fulminant neuropathy with severe quadriparesis occurring in patients with leukemia being treated with vincristine (and who do not have coexistent Charcot-Marie-Tooth disease) is more likely due to Guillain Barre syndrome than to vincristine neurotoxicity. PMID- 11054981 TI - Detrimental effects of high-dose dexamethasone in severe, refractory, HIV-related thrombocytopenia. AB - OBJECTIVE: To report a case of HIV-related thrombocytopenia in which high-dose dexamethasone was ineffective and immunologically detrimental. CASE SUMMARY: A 39 year-old white man with persistent, severe, HIV-1-related thrombocytopenia was admitted for epistaxis, bleeding gums, petechiae, and bruising. Previous unsuccessful attempts to reverse the thrombocytopenia included zidovudine, prednisone, vincristine, interferon alfa, and intravenous immune globulins. Based on previous anecdotal reports of the effectiveness of high-dose dexamethasone in refractory, HIV-related thrombocytopenia, we instituted treatment with intravenous dexamethasone 40 mg/d for four sequential days every 28 days. After three cycles of therapy, the platelet count remained < 15 x 10(9)/L; however, the CD4+ lymphocyte count decreased progressively from 1447 x 10(6)/L at baseline to 560 x 10(6)/L three months after the third cycle. Due to persistent, severe thrombocytopenia and bleeding, the patient underwent splenectomy, resulting in normalization of the platelet count. DISCUSSION: High-dose dexamethasone has been proposed as treatment for patients with immune thrombocytopenia as an alternative to chronic oral corticosteroids and claimed to be associated with better effectiveness and fewer adverse effects. The results of this treatment in our patient show that this regimen may not only be ineffective, but may also be immunologically detrimental in HIV-infected patients. Although the deterioration of the immunologic status of our patient cannot be fully attributed to high-dose dexamethasone based on the Naranjo scale, the previous long-lasting stability of CD4+ cells and the temporal relationship of a decrease in the CD4+ cell count coinciding with administration of high-dose dexamethasone suggest a causative role of the treatment. CONCLUSIONS: A possible cause-effect relationship between the treatment and the decrease in the CD4+ cell count suggests that the use of high-dose dexamethasone may not be justified in patients with severe, HIV-related thrombocytopenia. PMID- 11054982 TI - Hypothyroidism and depression: a therapeutic challenge. AB - OBJECTIVE: To describe a patient with longstanding depression and hypothyroidism who had marked mood improvement only after triiodothyronine (T3) was added to her thyroxine (T4) replacement therapy. CASE SUMMARY: A 50-year-old white woman had a long history of depression and documented hypothyroidism since 1991. Despite treatment with T4 with dosages up to 0.3 mg/d, she continued to be depressed, have symptoms of hypothyroidism, and have a persistently elevated thyroid stimulating hormone concentration. Addition of a low dose of T3 to her regimen resulted in significant mood improvement. DISCUSSION: The relationship between hypothyroidism and depression is well known. It is possible that this patient's long history of depression may have been a consequence of inadequately treated hypothyroidism, due either to poor patient compliance or resistance to T4. Nevertheless, her depression responded to addition of a low dose of T3 to her regimen. This case emphasizes the importance of screening depressed patients for hypothyroidism. Her clinical course also suggests that depression related to hypothyroidism may be more responsive to a regimen that includes T3 rather than to replacement with T4 alone. This is consistent with the observation that T3 is superior to T4 as adjuvant therapy in the treatment of unipolar depression. CONCLUSIONS: Depressed patients should be screened for hypothyroidism. In hypothyroid patients, depression may be more responsive to a replacement regimen that includes T3 rather than T4 alone. Therefore, inclusion of T3 in the treatment regimen may be warranted after adequate trial with T4 alone. PMID- 11054983 TI - High-flux hemodialysis without hemoperfusion is effective in acute valproic acid overdose. AB - OBJECTIVE: To report a case of valproic acid overdose treated successfully with high-flux hemodialysis without the addition of charcoal hemoperfusion. CASE SUMMARY: A 25-year-old white woman with a history of multiple suicide attempts and schizophrenia presented after ingesting an unknown amount of valproic acid. She became comatose and developed hypotension and lactic acidosis as valproic acid concentrations increased to > 1200 micrograms/mL (therapeutic concentration 50-100). High-flux hemodialysis was performed for four hours; the calculated elimination rate constant (kel) during the procedure was 0.2522 h-1 with a half life (t1/2) of 2.74 hours compared with posthemodialysis kel of 0.0296 h-1 and t1/2 of 23.41 hours, suggesting that high-flux hemodialysis effectively eliminates valproic acid. The patient's hemodynamic status and mental function improved in conjunction with the acute reduction in valproic acid concentrations. Her subsequent hospital course was complicated only by transient thrombocytopenia. DISCUSSION: Most literature reports of valproic acid overdose have described the use of charcoal hemoperfusion alone or in combination with hemodialysis to accelerate valproic acid clearance at toxic concentrations. However, the pharmacokinetic properties of valproic acid indicate that hemodialysis alone would be effective therapy for an acute valproic acid overdose. CONCLUSIONS: We suggest that toxic concentrations of valproic acid can be effectively reduced with high-flux hemodialysis without the addition of charcoal hemoperfusion and its attendant risks. PMID- 11054984 TI - Metronidazole-associated pancreatitis. AB - OBJECTIVE: To report a case of acute reversible pancreatitis associated with metronidazole-treated aspiration pneumonia. CASE SUMMARY: A 61-year-old white woman requiring coronary artery bypass surgery developed acute pancreatitis following treatment with metronidazole for suspected postsurgical aspiration pneumonia. The patient developed moderate to severe bilateral upper quadrant abdominal pain; laboratory studies revealed elevated amylase and lipase concentrations four days following the initiation of metronidazole therapy. After discontinuation of metronidazole, the patient's abdominal pain subsequently improved, and both amylase and lipase concentrations immediately declined and were within normal limits within one week. DISCUSSION: An acute attack of pancreatitis is characterized by moderate to severe abdominal pain that may radiate to the back, accompanied by increased concentrations of pancreatic enzymes and few morphologic changes in the pancreas. Metronidazole is reported as having a probable association with acute pancreatitis, although the mechanism of drug-induced pancreatitis is not known. One speculative mechanism of metronidazole-induced pancreatitis is that, under aerobic conditions, metronidazole may undergo redox cycling and yield hydrogen peroxide, superoxide, and other free radicals. Such redox-active compounds are toxic to pancreatic beta cells, and oxygen-centered free radicals have been implicated in the induction of pancreatitis. Other suggested mechanisms include immune-mediated inflammatory response, pancreatic duct constriction, and metabolic effects. CONCLUSIONS: Very few cases of metronidazole-associated pancreatitis have been reported, and the long-term sequelae are unknown. However, if metronidazole or any other drug is suspected as the causative agent in pancreatitis, it should be discontinued and rechallenge should be avoided. PMID- 11054985 TI - Treatment of chronic hepatitis C virus infection. AB - OBJECTIVE: To review the literature on the treatment of chronic hepatitis C virus (HCV) infection. DATA SOURCES: MEDLINE search (1986-December 1999) using key words such as HCV, hepatitis, non-A and non-B hepatitis, as well as terms regarding treatment during that time period. DATA SYNTHESIS: HCV infection was initially treated with interferon monotherapy, but only a minority of patients responded to long-term therapy. A higher rate of response in both interferon naive patients and interferon-relapsers has been achieved by using the combination of interferon and ribavarin. Other treatment regimens including high dose interferon protocols, ursodeoxycholic acid, amantadine, and nonsteroidal antiinflammatory drugs have been less promising. Many alternative therapies are being investigated. CONCLUSIONS: HCV infection is a major public health problem. It is now possible to achieve a cure in nearly 50% of the patients with this infection. Many additional therapies are being evaluated in order to achieve a higher cure rate. PMID- 11054986 TI - Use of epidural corticosteroids in low back pain. AB - OBJECTIVE: To review the literature regarding the safety and efficacy of epidural corticosteroid injections in the treatment of low back pain (LBP) of various etiologies. DATA SOURCES: A MEDLINE search (1966-February 1999) of English language literature pertaining to the use of epidural corticosteroids in LBP was performed. Additional literature was obtained from reference lists of pertinent articles identified through the search. STUDY SELECTION AND DATA EXTRACTION: All articles were considered for the review. The clinical trials included are those that enrolled a larger patient population and were primarily controlled clinical trials. The authors selected pertinent information for further discussion. DATA SYNTHESIS: Nerve root compression and inflammation are thought to be factors contributing to LBP. Corticosteroids act to decrease inflammation and may be of benefit in relieving LBP, especially if administered directly to the affected area via the epidural route. However, data on the efficacy of various corticosteroid agents administered via this route are limited. In addition, there have been reports of significant adverse reactions, thought to be due primarily to the preservative components in the corticosteroid preparations. Therefore, the role of these agents in the therapy of LBP remains to be determined. CONCLUSIONS: Based on the studies reviewed, epidural corticosteroids may be an effective treatment for LBP. Their use is warranted in patients who have failed conservative therapy. Although they contain preservatives, it appears that these agents are relatively safe and do not cause significant neurotoxicities. PMID- 11054987 TI - Clinical pharmacology of encapsulated sustained-release cytarabine. AB - BACKGROUND: The therapeutic effectiveness of chemotherapy is often limited by the inability to sustain cytotoxic concentrations at the tumor site. Cytarabine liposome injection (DepoCyt), a sterile, injectable suspension of the antimetabolite cytarabine, encapsulated into multivesicular, lipid-based particles, has been developed to improve the treatment of neoplastic meningitis (NM) through sustained release of cytarabine. OBJECTIVE: To review the pharmacokinetics, efficacy, and safety of intrathecal DepoCyt for the treatment of NM secondary to lymphoma or solid tumors. RESULTS: In preclinical and clinical studies, DepoCyt markedly extended the duration of tumor exposure to cytotoxic concentrations of cytarabine compared with administration of unbound cytarabine. Data from recent clinical studies demonstrate that DepoCyt improves complete response rates among patients with NM secondary to lymphoma. Trends in time to neurologic progression and median survival also favored DepoCyt over unbound cytarabine in these studies. Data have also been presented that suggest that patients with NM secondary to solid tumors benefit more from DepoCyt than from conventional treatment approaches. Chemical arachnoiditis (i.e., headache, fever, nausea, vomiting) was common in patients receiving DepoCyt, however, symptoms were manageable with oral dexamethasone. CONCLUSIONS: Encapsulation of cytarabine into liposomes for sustained release prolongs tumor exposure to cytotoxic concentrations of cytarabine, which may improve therapeutic efficacy in patients with NM secondary to lymphoma or solid tumors. PMID- 11054988 TI - Mild cognitive impairment: emerging therapeutics. AB - OBJECTIVE: To present a general overview of the etiology, definition, and prevalence of mild cognitive impairment (MCI), as well as outline possible treatment strategies. DATA SOURCES: A MEDLINE search was conducted for relevant references generated from 1990 to 2000 concerning MCI, mild to moderate Alzheimer disease (AD), and therapeutic strategies. Several books were also used in the compilation of data for this review, as well as the authors' experience in designing and conducting MCI trials. DATA EXTRACTION: All of the references listed were assessed, and all relevant information was included in this review. DATA SYNTHESIS: Forgetful individuals most likely to develop AD have a condition known as MCI previous to their development of dementia. This condition is hallmarked by memory impairment that is abnormal for the individual's age and educational level. While not all individuals with MCI develop AD, it is apparent that the condition can serve as a potential marker for early onset of AD. CONCLUSIONS: As many clinicians can attest, occasional forgetfulness is a common aspect of the aging process. Eventually, however, a large portion of forgetful individuals, especially those with MCI, will be diagnosed with AD or some other form of dementia. Indeed, many researchers have suggested that MCI should be regarded as incipient AD and that these individuals would benefit from drug therapy. Thus, MCI screening may be beneficial in terms of both early AD intervention and perhaps even AD prevention. PMID- 11054989 TI - Transdermal nitroglycerin for the prevention of intravenous infusion failure due to phlebitis and extravasation. AB - OBJECTIVE: To evaluate the clinical use of transdermal nitroglycerin for the prevention of intravenous infusion failures due to phlebitis or extravasation. DATA SOURCES: Clinical literature was accessed through a MEDLINE search (1966-May 2000). Key search terms included nitroglycerin, transdermal, phlebitis, extravasation, intravenous, and infusion. DATA SYNTHESIS: Common and serious consequences of intravenous therapy include the occurrence of postinfusion phlebitis and failure to maintain intravenous therapy due to intravenous fluid extravasation. Transdermal application of nitroglycerin has been evaluated as a treatment and preventive method for intravenous infusion failures related to phlebitis or extravasation. An evaluation of studies focusing on transdermal nitroglycerin in the prevention of infusion failures due to phlebitis or extravasation was conducted. CONCLUSIONS: Use of transdermal nitroglycerin as a prophylactic measure for intravenous infusion failures is a therapeutic option for patients requiring long-term intravenous therapy (i.e., > 50 h). PMID- 11054990 TI - Vitamin C in the prevention of nitrate tolerance. AB - OBJECTIVE: To evaluate the efficacy of vitamin C in preventing the development of nitrate tolerance. DATA SOURCES: A MEDLINE search (from 1966 to July 2000) was conducted to identify relevant articles, with additional references obtained from the bibliographies of these articles. DATA SYNTHESIS: One possible mechanism of nitrate tolerance involves superoxide-induced deactivation of nitric oxide, providing the rationale for the use of antioxidants. Most published research concerning deactivation of nitric oxide has involved vitamin C; a summary of this information is presented here. CONCLUSIONS: Preliminary studies seem to support the role of vitamin C in attenuating the development of nitrate tolerance. Considering these findings, larger, long-term trials are necessary to further establish the role of vitamin C in this situation. PMID- 11054991 TI - Costs related to inappropriate use of albumin in Spain. AB - BACKGROUND: Albumin has been used in various treatments for > 50 years, but, recently, its use in clinical practice has become very controversial. OBJECTIVE: To assess the use of albumin in clinical practice in the public hospital setting in Andalucia, Spain, focusing on the economic repercussions of the inappropriate use of albumin. METHODS: Multicentered observational study in 22 public hospitals in which all patients receiving albumin (from start to conclusion of treatment) were assessed during a five-month period on three predetermined, nonconsecutive days. The clinical indications for albumin were evaluated on the basis of Guidelines, a consensus document created by a multidisciplinary team for dissemination by the Governmental Health Authority to all hospitals within its purview. The data were abstracted from the patient case report forms by the pharmacist selected to compile the data in each of the participating hospitals. RESULTS: A total of 242 forms reporting the use of 62,282 g of albumin were evaluated. The most frequent prescribing motives were nutritional intervention (23%), paracentesis in cirrhotic patients (19%), and radical surgery (11%). Only 59 prescriptions (24%), corresponding to 14,539 g of albumin (23%), were considered appropriate. The total cost of albumin therapy for the 242 cases was $183,796 (US$); $42,905 (23%) of this figure was the cost of appropriate use of albumin and $140,891 (77%) was the amount related to inappropriate use. CONCLUSIONS: Evaluated against model guidelines, the use of most of the albumin, deemed clinically necessary by the prescribers, was considered unnecessary or inappropriate. Hence, institutions need to define and implement guidelines that focus on responsible use of such agents in an increasingly cost-conscious healthcare environment. PMID- 11054992 TI - Comment: cost-effectiveness of macrolides in lower respiratory tract infections. PMID- 11054993 TI - Comment: treatment of hyperhomocysteinemia with folic acid. PMID- 11054994 TI - Amitriptyline and somnambulism. PMID- 11054995 TI - Urticaria and respiratory distress due to porfimer sodium. PMID- 11054996 TI - [Endocoronary radiotherapy. A main therapeutic overhang or a Sorcerer's Apprentice technique?]. PMID- 11054997 TI - [Endovascular treatment of aorto-iliac aneurysms. Made-to-measure endoprotheses increase the feasibility]. AB - The authors describe their experience of tailoring endoprostheses for endovascular treatment of aorto-iliac aneurysms with components available on the market. Between January 1996 and December 1999, 188 aorto-iliac aneurysms were treated by tailor-made endoprostheses using self-expanding Z stents made of stainless steel compiled with polyester ligatures and covered with standard commercially available polyester prostheses. These endoprostheses were implanted with an 18 to 24 Fr (usually 20 Fr) introducer and positioned by a surgical approach. This method allows construction of tubular, bifurcated, digressive or occlusive endoprostheses associated with an extra-anatomical bypass graft. It increased the number of endovascular procedures for aorto-iliac aneurysms in the authors' department. This number has been further increased by using endoprostheses with an uncovered proximal or distal stent for cases with particularly short or angled necks and by using hybrid endoprostheses with one or more extremities without a stent, allowing surgical suture of the anastomosis. The authors' results show that tailoring endoprostheses considerably increased the feasibility of endovascular treatment of aorto-iliac aneurysms, even in unselected patients whilst providing an effectiveness and safety to justify the continuation of this experience. PMID- 11054998 TI - [Effect of age on the prognosis of infectious endocarditis]. AB - The aim of this study was to evaluate the influence of age on the prognosis of infectious endocarditis. A retrospective study from 1987 to 1997 of 136 patients with infectious endocarditis on native, prosthetic valves or cardiac pacing catheter was performed. The outcome was analysed with the help of general practitioners. Two groups of patients were compared: 87 patients of 65 years of age or more (Group 1) and 49 patients under 65 years of age (Group 2). With a follow-up period of 5 years, the global mortality was 35%, but greater in Group 1 (p = 0.06). Cardiac failure was the main cause of death. The mortality was significantly higher in patients who were not operated (p < 0.002). The authors conclude that age of over 65 does not significantly worsen the prognosis of infectious endocarditis. The absence of surgery seems to be an indirect factor of a poor prognosis. Long-term follow-up of patients is necessary to diagnose and treat cardiac failure at an early stage and to consider referral for surgery. PMID- 11054999 TI - [Validation of the measurement of cardiac output by acoustic quantification in patients with severe congestive heart failure]. AB - The technique of acoustic quantification (AQ), because of its automatic detection of the contours, enables left ventricular volumes to be calculated in real time using the technique of disk summation. The objective of the study was to evaluate the reliability of cardiac output (CO) measurements obtained with AQ based on left ventricle volumes in patients with severe congestive heart failure. Seventeen patients, mean age 68 +/- 11 years, NYHA stage IV, in sinus rhythm and without significant valve regurgitation were enrolled prospectively. CO, measured simultaneously by 2-dimensional echocardiography (2DE), pulsed Doppler echocardiography (DOP) and AQ, was compared to the thermodilution technique (TD) data. CO, measured by AQ, was highly correlated with TD (r = 0.875; p < 0.001) with a small bias (-0.05 l/min). DOP and 2DE were also well correlated with TD (r = 0.835 and r = 0.701, respectively). Concerning ventricular volume measurement, AQ was well correlated with 2DE (for telediastolic, r = 0.892, and telesystolic volumes, r = 0.874). However, telesystolic (bias, +36.6 +/- 35 ml) and telediastolic volumes (bias, +35.6 +/- 35 ml) were overestimated. We conclude that AQ is a reliable technique, able to estimate CO precisely in patients with severe congestive heart failure. On the other hand, both telesystolic and telediastolic volumes were overestimated. PMID- 11055000 TI - [Curative radiofrequency ablation of paroxysmal junctional tachycardia in patients over 70 years of age. A multicenter study]. AB - Curative radiofrequency ablation of the reentry circuit of paroxysmal junctional tachycardia is a relatively common method of treating this condition. The aim of this study was to determine whether the age of the patients should be taken into account for assessing the indication. The study population was 178 patients aged 18 to 86 years (average 56 +/- 19 years), who had paroxysmal junctional tachycardia and normal interatrial ECGs. One hundred and thirty-five patients were under 70 years of age (Group I) and 43 were over 70 (Group II). No significant differences in the mechanism of reentry, which was intranodal in 67% of cases, in the risk of immediate complications (11%) or in recurrence of tachycardia were observed between the two groups. Functional improvement was more spectacular in Group II with regression of the symptoms of associated cardiac disease and, above all, in reduction in the number of hospital admissions. The common association of cardiac disease and other pathologies in Group II should however lead to more careful management and follow-up of the more elderly patients. The authors conclude that radiofrequency ablation of paroxysmal junctional tachycardia in the over 70s is feasible and often provides better clinical results than observed in younger patients. PMID- 11055001 TI - [Vasodilator effects of bradykinin on the resistive circulation of the forearm of coronary patients]. AB - This study was undertaken to determine the vascular effects of bradykinin and its modes of action on the resistive circulation of the forearms of coronary patients and healthy volunteers. Two groups were studied: Group I comprising 8 coronary patients with normal left ventricular function and Group II with 8 healthy volunteers. The method of measurement of forearm blood flow was occlusive venous plethysmography with a mercury strain gauge. The vasodilatory response of the two groups to local arterial perfusion of acetylcholine (40 and 80 micrograms/min), bradykinin (10, 30, 100 pmoles/min), then the association of L-Ng-monomethyl-L arginine (L-NMMA) at 8 microG/min, an inhibitor of the synthesis of nitric oxide, with acetylcholine and bradykinin. Five subjects of the control group received oral aspirin (250 mg/day) for one week before the study. Acetylcholine and bradykinin increased forearm flow in a dose-dependent manner but the increase was significantly less in coronary patients than in the healthy volunteers (p < 0.05). The L-NMMA inhibited the vasodilatory response to acetylcholine by about 40% in the two groups but had no significant effect on the vasodilatation induced by bradykinin. Aspirin had no effect on the vasodilatation induced by acetylcholine or bradykinin. These data show that the vasodilatory response to bradykinin is decreased in coronary patients and suggest that nitric oxide is not the second main messenger of bradykinin in the resistive circulation of the forearm. PMID- 11055002 TI - [Qualitative and quantitative descriptions of the mechanisms of action of the angioplasty balloon on coronary stenosis. An endovascular ultrasonic study]. AB - The barotrauma induced by inflation of the balloon on an obstructive atherosclerotic plaque induces several distinct, complex effects. The object of this study was to describe these mechanisms based on endovascular echographic sections of coronary stenoses before and after balloon angioplasty. The 32 lesions analysed were richly cellular (81.2%) and associated with calcifications in 31% of cases. The remodelling index before angioplasty was used to individualize a majority of lesions with chronic arterial constriction (56.2%). The modelling of the plaque (dilatation or constriction) had no effect on the final luminal result. Global analysis of the endoluminal gain (4.38 +/- 2.28 mm2) showed that it was mainly due to reduction of plaque surface (78.2% of gain) without prejudging the mechanism, and less due to expansion of the global arterial surface (21.8% of the gain). The type of remodelling affected the mechanisms of action of balloon angioplasty. Dissection was present in 53.1% of cases. Fragmentation of the plaque with embolisation is a common phenomenon (28% of cases). The authors conclude that there are four mechanisms which coexist: 1) Reduction of plaque thickness cannot physically correspond to simple compression of tissue. The plaque is redistributed longitudinally. 2) Arterial expansion only plays a minor part in endoluminal gain. 3) Plaque rupture is directly related to the acute increase in wall stress often exceeding the thresholds of rupture of its components. Finally, 4) embolisation by parietal fragmentation, a mechanism often unknown or ignored which plays an essential part in the potentially deleterious effects. PMID- 11055003 TI - [Isolated tricuspid valve replacement. Long-term results]. AB - The purpose of this study was to examine the early and late results in 29 patients who underwent 32 (6 mechanical and 26 bioprostheses) isolated tricuspid valve replacement (TVR) from a total of 79 TVR and 375 tricuspid annuloplasties performed at the Montreal Heart Institute, between January 1978 and January 1998. Patients' ages ranged from 25 to 70 years (mean 48 years), and 62% were females. Twenty-seven patients (84%) were in New York Heart Association (NYHA) functional class III and IV. Previous valve surgery had been performed in 22 patients (69%) of which 9 had undergone TVR. Postoperatively, permanent pacemaker was implanted in 9 patients (28%), and immediate reoperation was required in 2 patients because of bleeding. Mean follow-up period was 67.7 months (93% complete). Serial echocardiography showed 3 prosthesis dysfunctions, leading to a second replacement in 2 patients at 12.8 and 7.7 years after initial surgery. All but three patients showed an improvement of their NYHA class. Hospital mortality occurred in 6 patients (19%) and 7 patients died during late follow-up: mean 38.1 months after surgery, including one valve-related death (mechanical valve thrombosis). Actuarial survival rate of all patients was 63% after 5 years, and 47% after 10 years. Isolated TVR remains a high-risk procedure. Most survivors, however, should expect a better quality of life with improvement in their NYHA class. PMID- 11055004 TI - [Use of radiotherapy in cardiovascular disease. Radiophysical basis, current results, indications and perspectives]. AB - Restenosis is the main limitation of percutaneous angioplasty, especially in vessels of small diameters such as the coronary arteries, the femoro-popliteal and tibial-peroneal arteries and the arterio-venous dialysis grafts. The extensive use of tents has not entirely prevented its occurrence, whereas treating in-stent restenosis gives even more uncertain results. Endovascular radiotherapy has emerged over the past few years as a promising approach to both prevent and cure it. The analogy between the tumour-like cellular proliferations observed in post-angioplasty restenosis and tumour processes prompted pioneering works to study the effect of ionizing radiations in animal models of arterial restenosis. The demonstrated feasibility, tolerance and efficacy of this approach lead to test this strategy in humans. The results of 3 recently presented randomized double-blind trials in the treatment of coronary in-stent restenosis have been so promising that endovascular brachytherapy might now be considered the treatment of choice in this indication. Other randomized trials are currently carried out to test whether endovascular brachytherapy may prevent restenosis in coronary and femoro-popliteal arteries as well as in hemodialysis shunts. In the present review, we describe the basics of the biological effects of ionizing radiations, the technical modalities to deliver endovascular radiations, our current knowledge about their effects on the vascular wall and the restenosis mechanisms, and the results of the first clinical studies. Finally, we address the remaining problems in the use of endovascular curietherapy and question the promises and challenges of its clinical application. PMID- 11055005 TI - [Measurement of carotid artery intima-media thickness. Analysis and reproducibility]. AB - The intima-media thickness of the carotid artery is a cardiovascular risk factor, especially in hypertensive, diabetic or dyslipidemic patients. Very accurate tools of measurement are essential in order to optimise the analysis of the severity of the vascular lesions. Present methods use computer programmes for ultrasonic image enhancement and have already been shown to be very reproducible. However, there is still room for improvement in the detection of lesions at their initial stage of development and in the follow-up of their progression with or without antihypertensive or lipid lowering drugs. PMID- 11055006 TI - [Pericardial effusion revealing cardiac amyloidosis in the course of rheumatoid arthritis]. AB - Pericardial effusion is common in patients with rheumatoid arthritis. It is essentially a sign of pericardial involvement of the rheumatoid disease, but viral, bacterial and especially tuberculous pericarditis must not be excluded. Pericardial amyloidosis of the AA type is much less common and difficult to diagnose before cardiac biopsy even in cases of myocardial amyloidosis, as in the reported case, in which the classical association of microvoltage on the ECG and myocardial hypertrophy on echocardiography was absent. The absence of myocardial uptake of technetium-labelled pyrophosphates at myocardial scintigraphy and the absence of a restrictive profile on cardiac gamma-angiography were not suggestive of the diagnosis of amyloidosis. Pericardial and endomyocardial biopsy, justified by the negativity of the preceding investigations, provided an accurate histological diagnosis, a prognostic evaluation and was also useful for guiding management. PMID- 11055007 TI - [Paroxysmal atrioventricular block during coronary revascularization with ultrasound. A case report]. AB - The authors report the case of advanced atrioventricular block during a procedure of right coronary revascularisation with ultrasound. The different possible physiopathological mechanisms are discussed. Although no formal conclusions can be drawn, certain features suggest a paroxysmal nodal conduction defect secondary to transient parasympathetic stimulation by the ultrasound, a stimulation triggered by a Bezold-Jarish type of cardiac reflex or by direct local excitation of vagal fibres of the atrioventricular node. PMID- 11055008 TI - [Right atrioventricular metastasis of a myxoid liposarcoma. Case report and al review of the literature]. AB - Malignant tumours of the heart and pericardium are rare. Secondary tumours are 20 to 40 times more common than primary tumours. The authors report the case of a cardiac metastasis of a myxoid liposarcoma of the inguinal region. Surgical ablation with chemotherapy significantly improved the patient' clinical and haemodynamic status. A review of the literature shows that postoperative survival does not exceed 2 years, but the prognosis is better when chemotherapy is associated with surgery. Transoesophageal echocardiography is an excellent diagnostic imaging method. It provides a better evaluation of the tumour and its location, compared to thoracic CT scan and transthoracic echocardiography. PMID- 11055009 TI - Transfer of lead via placenta and breast milk in human. AB - The mean lead levels in the maternal blood, cord blood, breast milk and placental tissue, were 0.63 mumol/L (13.2 micrograms/dL), 0.33 mumol/L (6.90 micrograms/dL), 4.74 micrograms/L and 0.86 mumol/kg (17.85 micrograms/100 g) respectively for 165 parturient women occupationally non-exposed to lead in 2 hospitals in Shanghai. No significant difference was found between maternal age groups for these indicators. However, the lead levels in the cord blood and breast milk increased with the lead level in the maternal blood, with coefficient of correlation of 0.714 (P < 0.0001) and 0.353 (P < 0.01) respectively. The mean concentration of lead in breast milk for 12 occupationally lead exposed women was 52.7 micrograms/L, which was almost 12 times higher than that for the occupationally non-exposed population. These results suggested that transfer of lead via placenta prenatally and breast milk postnatally were possible and might pose a potential health hazard to the fetuses and the neonates. PMID- 11055010 TI - Acute inhalation toxicity study of 2-fluoroacetamide in rats. AB - One of the most potent rodenticides is 2-fluoroacetamide (2-FA). Toxicity of this chemical is well documented. However, its inhalation toxicity data is not available in the literature. Hence, acute inhalation toxicity study was carried out by exposing male and female rats to aerosols of 2-FA at different concentrations for 4 h in a dynamically operated whole body inhalation exposure chamber. During and after the inhalation exposure the rats were less active, and showed mild tremors and convulsions. At higher concentrations the rats died after 2-3 days. The estimated 4-h LC50 for male and female rats was 136.6 and 144.5 mg.m-3 respectively. Exposure to 0.7 LC50 for 4 h duration showed an increase in the liver weight of male and female rats 7 days after exposure. Various haematological and biochemical variables determined were within the normal limits. However, histological findings showed injured lung as indicated by desquamation and necrosis of the epithelium of the respiratory tract. Marked hypertrophy of hepatocytes displaying strong acidophilic granulated cytoplasm was observed. Focal dilatation of renal proximal tubules in kidney with cytoplasmic vacuolation, and irregularly placed pyknotic nuclei were seen. The present study shows that 2-FA is a highly toxic chemical through the inhalation route based on the LC50 value. Consequently necessary precautions should be taken during its handling. PMID- 11055011 TI - The identification and toxicity of the microcystin in the waterbloom obtained from an eutrophied lake. AB - The water of "J" lake has been seriously eutrophied; concentration of total nitrogen (TN), total phosphorus (TP) and chlorophyll a were all far above the 3rd level of the National Standard of Ground Water of China. The concentration of microcystin (MCYST) of the water at one site (M) was 1865 micrograms/l. There were 2.36 micrograms MCYST-LR per mg dry waterbloom powder. PMID- 11055012 TI - Effect of dietary fatty acids on colon tumorigenesis induced by methyl nitrosourea in rats. AB - To study the effect of dietary fatty acid on the colon tumorigenesis induced by methyl nitrosourea in rats, male SD rats were fed five semi-synthetic diets composed of different proportions of beef tallow, soybean oil, alkana oil, corn oil and fish oil for 180 days. The experimental groups were injected with a solution of methyl nitrosourea in phosphate buffer intraperitoneally once a week for six weeks. The control groups were injected with phosphate buffer solution only. The incidence of colon cancer, the average volume of the tumors, proliferation cell nuclear antigen, cell kinetics, membrane lipid fluidity, alkaline phosphatase activities and the content of prostaglandin E2 in colon mucosa and the fatty acid of testis pad fat were measured at the end of the experiment. The results showed that the incidence of colon cancer and the average volume of tumors in animals fed with diets composed mainly of beef tallow, soybean oil or alkana oil were significantly higher than those that were fed fish oil. The most effective anticancer diet in our study contained saturated fatty acid, monounsaturated fatty acid and polyunsaturated fatty acid of fish oil in the proportion of 13.9%, 16.4% and 68.8% respectively. Inhibition of colon tumorigenesis appeared to be related to the regulation of membrane lipid fluidity, and a decrease in the proliferation of cell nuclear antigen in colon cells. In addition, a decrease was noted in the number of cells in S phase and alkaline phosphatase activity, along with inhibition of arachidonic acid products and a corresponding decrease in the amount of prostaglandin E2. PMID- 11055013 TI - Neutral red retention by earthworm coelomocytes: a biomarker of cadmium contamination in soil. AB - The earthworm Metaphire posthuma were used as a model to assess the toxic potential of cadmium incorporated into the soil by environmental or human activities. The retention period of neutral red in the lysosomes of the coelomocytes was used as a biomarker. The viability of harvested coelomocytes by a non-invasive extrusion protocol was 93% with no alteration by the dye during experimentation. The control cells retained dye for 119 and 121 min in normal soil and KCl, respectively, whereas a linear decline in the retention time in the treated earthworm coelomocytes was observed. This illustrated that the presence of cadmium caused damage to the lysosomes of the coelomocytes. PMID- 11055014 TI - Effect of nicotinamide on 12-O-tetradecanoyl-phorbol-13-acetate exposed mouse skin endonuclease activity and DNA synthesis. AB - Nicotinamide (NA), a relatively nontoxic compound, has been shown to inhibit tumor development, induce differentiation, increase the sensitization of the anticancer drug resistant cancer cells and is being used in different skin ailments. But there are not many reports on its mechanism of action. Here we report that NA induced endonuclease activity. This endonuclease induction by NA appeared to be dose dependent and a function of time. As evident by the use of modifiers of DNase I, this endonuclease appeared to be like DNase type I. Increased [3H] thymidine incorporation in DNA in the presence of NA is possibly a consequence of increased 3-OH' nicks due to increased DNA fragmentation by increased endonuclease activity. The present results would be of help in the better understanding of the mechanism of NA action and its improved use in cancer control. PMID- 11055015 TI - Heroin abuse and nitric oxide, oxidation, peroxidation, lipoperoxidation. AB - To further reveal the risks of heroin abuse to human body, and to determine the injuries of oxidation, peroxidation and lipoperoxidation induced by nitric oxide and other free radicals to heroin abusers, we determined and compared plasma values of lipoperoxides (LPO), nitric oxide (NO), vitamin C (VC), vitamin E (VE), beta-carotene (beta-CAR) and erythrocyte values of LPO, superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) in 114 heroin abusers and 100 healthy volunteers. Using linear regression and correlation as well as stepwise regression and correlation, we also analyzed the effect of the abusing duration, and daily abusing quantity on the above-mentioned biochemical parameters in the heroin abusers. The results showed that, compared with the healthy volunteer groups, the average plasma values of LPO, and NO, and the average erythrocyte value of LPO in the heroin abuser group were significantly increased (P < 0.0001), and the average plasma values of VC, VE, and beta-CAR and the average erythrocyte values of SOD, CAT, and GSH-Px were significantly decreased (P < 0.0001). Analysis of linear regression and correlation showed that with prolonged heroin abusing and with increased daily quantity in the heroin abusers, the plasma values of LPO, and NO, and the erythrocyte value of LPO were gradually increased (P < 0.001), whereas the plasma values of VC, VE, and beta CAR and the erythrocyte values of SOD, CAT, and GSH-Px were gradually decreased (P < 0.001). Analysis of stepwise regression and correlation indicated that the plasma values of NO, VC and VE were closely correlated with the abusing duration and daily abusing quantity. These results indicate that the balance between oxidation and antioxidation in the heroin abusers was seriously disturbed, and the injuries induced by nitric oxide and other free radicals, through oxidation, peroxidation and lipoperoxidation to the bodies of heroin abusers exacerbated. It is therefore necessary that in abstaining from heroin dependence, the heroin abusers should acquire sufficient quantities of antioxidants such as VC, VE and beta-CAR. PMID- 11055016 TI - Estimation of burden of disease for smear-positive pulmonary TB and its infectivity. AB - The study investigated the burden of smear-positive pulmonary TB and its infectivity, using DALY (Disability-Adjusted Life Year) as an indicator. An assumed cohort of 2000 cases was set up based on the age-specific incidence of 794 newly registered smear positive cases of TB in Beijing in 1994. Prognostic trees and model diagrams of infectivity under natural history and DOTS (Direct Observed Treatment, Short-course) strategy were established according to the epidemiological evidence. The results show that 29.6% of DALYs would be neglected if the burden caused by the infectivity was not considered. The results also show that DOTS strategy may reduce 97.3% of the number of potential cases infected, 92.9% of DALYs related to TB-patients themselves, and 99.9% of DALYs caused by TB's infectivity as well. PMID- 11055017 TI - The prevalence of NIDDM and IGT and related factors among residents in some areas of Hubei Province, China. AB - The epidemiological survey of prevalence of NIDDM (non-insulin dependent diabetes mellitus) and IGT(impaired glucose tolerance) was conducted among 9450 residents aged 25-70 in some areas of Hubei Province, China. The results show that NIDDM and IGT prevalences are 2.62% and 4.48%, respectively. There is no significant difference between male and female (P > 0.05). The NIDDM prevalence in cities is slightly higher than that in countryside, but the difference is not significant (P > 0.05). However, the IGT prevalence in city is significantly higher than that in countryside (P < 0.01). The prevalence of both NIDDM and IGT is increasing along with the age of the population. It is also significantly related to the family history of NIDDM, hypertension, and high body mass index (BMI). By using stepwise logistic regression to analyse the risk factors of NIDDM, age (OR = 1.86), BMI(OR = 2.69), family history (OR = 2.84) and hypertension (OR = 2.23) entered the model (significance level is alpha = 0.05). PMID- 11055018 TI - National diabetes month: podiatric care of the diabetic foot. PMID- 11055019 TI - The Doppler probe for planning septofasciocutaneous advancement flaps on the plantar aspect of the foot: anatomical study and clinical applications. AB - A 5-MHz Doppler probe was utilized to identify the perforating septofasciocutaneous vessels on the plantar aspect of the right and left feet of 10 healthy subjects. Each audible perforator was marked, and each foot was photographed, scanned into a personal computer, and standardized to 8 inches high by 4 inches wide. All 20 feet were then stacked together to create a composite average of all markings. Loupe aided (2.5 x magnification) dissection of a latex cast of the perforating septofasciocutaneous vessels from a fresh frozen cadaveric foot revealed similar location and distribution as the composite average described above. The Doppler probe is capable of accurately identifying the septofasciocutaneous perforating vessels, thereby, creating a vascular map of the plantar aspect of the foot useful for precise planning of advancement flap coverage for full thickness defects. Two representative case examples are presented. PMID- 11055020 TI - Ostectomy for diabetic neuroarthropathy involving the midfoot. AB - Diabetic neuroarthropathy is a frequent complication of diabetes mellitus that results in instability of the foot, structural deformity, and soft-tissue breakdown secondary to increased plantar pressure. The midfoot is commonly involved in diabetic neuroarthropathy. Collapse of the medial, lateral, or both longitudinal arches may result in increased plantar pressures and subsequent midfoot ulceration. The majority of these wounds can be managed with local wound care, off-loading, and other forms of nonoperative care. Surgical intervention becomes necessary when the wound fails to heal with conservative measures. The authors performed a retrospective review of patients who underwent ostectomy for chronic or recurrent ulceration in the midfoot secondary to diabetic neuroarthropathy. The authors reviewed 27 procedures in 20 patients. There were 18 medial ulcers and 9 lateral ulcers. Wounds had resolved in 20 of 27 cases for 74% healing rate. The majority of failed procedures involved lateral column wounds (six of seven). Revisional surgery was required in five of the nine lateral column wounds for limb salvage. There was a statistically significant difference between the rate of complications by ulcer location (p = .00174). The rate of complications was significantly higher for lateral column ulcers. These results indicate that ostectomy is a reasonable option for medial column ulcers that fail nonoperative care. However, ostectomy for ulcers involving the lateral column is less predictable and failure often requires complex reconstructive soft tissue and osseous procedures for limb salvage. PMID- 11055021 TI - Barriers to podiatric care among diabetic patients in the San Francisco Bay area. AB - This study investigates the provision of general medical and foot care, the barriers to access for foot care, and the awareness of foot risks in an urban diabetic population. A survey composed of 26 questions was mailed to 2375 diabetic patients in the San Francisco Bay area who are members of the American Diabetes Association (ADA). Three hundred ninety-two surveys were returned for a response rate of 16%. Of the 392 respondents, 7 (1.8%) indicated that they were not receiving any medial care for their diabetes, with another 15 (3.8%) receiving general medical care from an alternative health care provider. Among the respondents, 87 (22%) did not have their feet examined by any health care provider. The remainder of the patients were receiving foot care from a health care provider with 191 (48.7%) under the care of a provider other than a podiatrist. Of those not receiving any foot care, 53 (61%) reported that they did not seek any pedal care because they do not have any apparent foot or leg problems. Another 12 (13.8%) indicated that they did not know whom to see for their lower extremity problems. Lack of insurance or inability to afford medical care was the main reason that prevented 7 (8%) of the patients from receiving routine foot care. With respect to the patient's knowledge of diabetes-associated foot disorders, the majority (72%-79%) knew that poor circulation, neuropathy, ulcers, painful leg and foot conditions, infection, and amputation were associated with diabetes. From all the surveyors, 106 (27%) reported that they were not advised or educated on the potential lower extremity complications of diabetes by their health care provider. The results of this study indicate that in an urban population of diabetic patients, all of whom were members of ADA, a significant number are not adequately educated on the importance of routine foot care. PMID- 11055022 TI - The natural history and longitudinal study of the surgically corrected clubfoot. AB - Surgical treatment for clubfoot has been largely directed at finding the best one stage operation for the resistant clubfoot. Eighteen patients with 27 clubfeet (average follow-up 11 years since first surgery; range, 3.5-24 years) were reviewed. More than one clubfoot operation was required in 56% of cases. Forty six percent were corrected after one surgery; 33% required a second surgery and 14% required a third operation. One patient with particularly severe feet required a fourth operation on each foot. The mean age at the time of surgery was 1.26 years, 5.12 years, and 8 years for the first, second, and third operations, respectively. The first operation consisted of a soft-tissue release. The second and third operations consisted of more extensive soft-tissue release and various rearfoot and forefoot procedures. Radiographic values revealed an AP talocalcaneal angle of 18 degrees, AP talo-first metatarsal angle of 6 degrees, lateral talocalcaneal angle of 29.6 degrees, lateral talo-first metatarsal angle of 15 degrees, and calcaneo-first metatarsal angle of 143 degrees. At follow-up all patients had adequate function as determined by personal interview and clinical examination. We conclude that correction of resistant congenital clubfoot often requires more than one surgery, not because of a "failed first operation," but due to dynamic muscle imbalances that may not be fully recognized in infancy and early childhood. Thus, the need for a second operation should not be perceived as a failure of the first, but as part of the natural history of congenital clubfoot. PMID- 11055023 TI - External constant tension expansion of soft tissue for the treatment of ulceration of the foot and ankle. AB - A clinical series of the management of 74 lesions in the foot and ankle region in 67 patients using a constant tension-approximating device is presented. The technical aspects of this methodology are detailed as they have been developed over the past 4 years. The indications, contraindications, and the requirements to obtain a successful result are discussed. The differences, as well as the possible advantages over other mechanical devices used in wound management, are discussed. The incidence and degree of complications are considered to be minimal when compared to both flap procedures and internal expansion technologies. PMID- 11055024 TI - Tuberculous osteomyelitis of the cuboid: a report of four cases. AB - Osteoarticular tuberculosis, although rare, has shown a resurgence in recent times, especially in immunocompromised patients. Involvement of the foot is infrequent, and the differential diagnosis is confusing, leading to diagnostic delays. We reviewed four cases of tuberculosis of the cuboid where the infection was limited to the bone without articular involvement. All four cases were adults and diagnostic delays were observed in all. Three of the cases had an osteolytic lesion on radiographs resembling a space-occupying lesion. Magnetic resonance imaging (MRI) or Computed tomography (CT) scans were helpful in three cases, and post-treatment MRI helped in evaluating outcome in one case. Antitubercular chemotherapy was sufficient to cause resolution of the lesion in three cases, while in one case surgical intervention was necessary. Since isolated osteomyelitis is usually seen only in the early stages of the disease process, early diagnosis and appropriate therapy are imperative to get good long-term results. Concomitant extraskeletal lesions are not always seen, nor is the organism cultured in a majority of the cases. Thus a high index of suspicion is mandatory in high-risk groups (immigrants, immunocompromised cases or those with history of contact); clinical and radiologic features, along with histopathologic evidence of granulomatous pathology should be sufficient to initiate therapy. PMID- 11055025 TI - Resection of talocalcaneal middle facet coalition. Interposition with a tensor fascia lata allograft: a case report. AB - Tensor fascia lata is utilized in the management of complex soft-tissue injuries and defects, but has not been described in the literature in the use of tissue interposition with resection of talocalcaneal middle facet coalitions. This article is a case presentation of a resection of a middle facet coalition with interposition of an allograft of tensor fascia lata. At 14 months postoperative follow-up, range of motion of the subtalar joint was noted to be 20 degrees, and without pain or crepitus. There was no radiographic evidence of degenerative changes in Chopart's joint. The patient returned to all routine and sports activities without pain. He was satisfied with the outcome of the procedure. PMID- 11055026 TI - Pedal consequences of rhabdomyolysis: a case report. AB - The authors present an unusual case that has been seldom reported in the literature. The clinical rhabdomyolysis syndrome, resulting from muscle disintegration, can be the result of numerous etiologic events and have serious sequelae. The pedal manifestations result from muscular and neurologic insult. The patient in this reported case survived rhabdomyolysis despite renal and hepatic failure requiring diuresis and hemodialysis. The only permanent long-term sequelae the patient suffered was flexible hammertoe deformities and parasthesias of all lesser digits of both feet. The follow-up is over 8 years. PMID- 11055027 TI - Osteochondral autogenous transplantation for osteochondritis dissecans of the ankle joint. PMID- 11055028 TI - Refusal of potentially life-saving blood transfusions by Jehovah's Witnesses: should doctors explain that not all JWs think it's religiously required? PMID- 11055029 TI - Care or custody? Ethical dilemmas in forensic psychiatry. PMID- 11055030 TI - 'If you pay, we'll operate immediately'. AB - OBJECTIVES: To study the attitudes of health care staff in four postcommunist countries towards taking gifts from their clients--and their confessed experience of actually taking such gifts. DESIGN: Survey questionnaire administered to officials including health care staff, supplemented by focus-group discussions with the general public. SETTING: Ukraine, Bulgaria, Slovakia and the Czech Republic. PARTICIPANTS: A quota sample of 1,307 officials including 292 health care staff, supplemented by stratified national random samples of 4,778 ordinary members of the public and in-depth interviews or focus-group discussions involving another 323. MAIN MEASUREMENTS: Explicit justifications and willingness to accept offers, reported frequency of offers, and personal confessions to accepting "money and expensive presents" as well as smaller gifts. RESULTS: Health care staff were far more inclined than the average official or public servant to accept "money or an expensive present" if offered, far more inclined to justify asking clients for "extra payments", and far more inclined to confess that they had actually taken gifts from clients recently. Judged by their own confessions, hospital doctors were only rivalled by traffic police and customs officials for taking money or expensive gifts from their clients. CONCLUSIONS: Poor pay does not explain why doctors so often took large gifts from their clients. Moral self justification, opportunity, and bargaining power are much more effective explanations. PMID- 11055031 TI - Priorities in care and services for elderly people: a path without guidelines? AB - The growing gap between demands and resources is putting immense pressure on all government spending in Sweden. The gap is especially apparent in care and services for elderly people in light of the rapid aging of the population. The article considers the decisions and priorities concerning resource allocation in the welfare sector in general and in elderly care in particular. The aim is to describe the political and administrative setting and to provide a conceptual structure that outlines the nature of the problem. Various levels of decision making are identified and discussed in the context of political accountability. Current transitions in elderly care are described with respect to service provision, marketisation, coverage rates, and eligibility standards. Basic principles of distribution are highlighted in order to clarify some central concepts of efficiency and justice, and a number of strategies for actual prioritising are identified. The article concludes with an endorsement of more conscious decisions in resource allocation. Existing knowledge and information concerning the effects of various strategies must be utilised, and the values and assumptions used for setting priorities must be made explicit. PMID- 11055032 TI - Randomisation and resource allocation: a missed opportunity for evaluating health care and social interventions. AB - Equipoise is widely regarded to be an essential prerequisite for the ethical conduct of a randomised controlled trial. There are some circumstances however, under which it is acceptable to conduct a randomised controlled trial (RCT) in the absence of equipoise. Limited access to the preferred intervention is one such circumstance. In this paper we present an example of a randomised trial in which access to the preferred intervention, preschool education, was severely limited by resource constraints. The ethical issues that arise when conducting randomised trials in health care are considered in the context of trials of social interventions. In health, education and social welfare, effective interventions are frequently limited due to budgetary constraints. Explicit acknowledgement of the need to ration interventions, and the use of random allocation to do this even in the absence of equipoise, would facilitate learning more about the effects of these interventions. PMID- 11055033 TI - Principles of justice in health care rationing. AB - This paper compares and contrasts three different substantive (as opposed to procedural) principles of justice for making health care priority-setting or "rationing" decisions: need principles, maximising principles and egalitarian principles. The principles are compared by tracing out their implications for a hypothetical rationing decision involving four identified patients. This decision has been the subject of an empirical study of public opinion based on small-group discussions, which found that the public seem to support a pluralistic combination of all three kinds of rationing principle. In conclusion, it is suggested that there is room for further work by philosophers and others on the development of a coherent and pluralistic theory of health care rationing which accords with public opinions. PMID- 11055034 TI - Should local research ethics committees monitor research they have approved? AB - The function of local research ethics committees is to consider the ethics of research proposals using human participants. After approval has been given, there is no comprehensive system in place to monitor research and ensure that recommendations are carried out. Some suggest that research ethics committees are ideally placed to fulfil this function by carrying out random monitoring of research they have reviewed. The health service guideline creating local research ethics committees is under review. This paper suggests that increasing the monitoring role of ethics committees in the present climate would be inappropriate. This is due to the large workload of the committees, their voluntary nature and the change a monitoring role might cause to the relationship between researcher and ethics committee, which might herald an increasing recourse to judicial review. A radical overhaul of the system would be necessary in order for ethics committees adequately to fulfil a monitoring function. PMID- 11055035 TI - The case for a new system for oversight of research on human subjects. AB - The increasing emphasis on evidence-based clinical practice has thrown into sharp focus multiple deficiencies in current systems of ethical review. This paper argues that a complete overhaul of systems for ethical oversight of studies involving human subjects is now required as developments in medical, epidemiological and genetic research have outstripped existing structures for ethical supervision. It shows that many problems are now evident and concludes that sequential and piecemeal amendments to present arrangements are inadequate to address these. At their core present systems of ethical review still rely on the integrity and judgment of individual investigators. One possible alternative is to train and license research investigators, make explicit their responsibilities and have ethics committees devote much more of their time to monitoring research activity in order to detect those infringing the rules. PMID- 11055036 TI - Metaphors, models and organisational ethics in health care. AB - Crucial to discussions in organisational ethics is an evaluation of the metaphors and models we use to understand the organisations we are discussing. I briefly defend this contention and evaluate three possible models: the current corporate model, an orchestrator model which puts hospitals in the same class as malls and airports, and a community model. I argue that the corporate and orchestrator model push to the background some important organisational ethics issues and bias us inappropriately towards certain solutions. Furthermore, I argue that the community model allows these to be more easily brought up. I also respond to the likely challenge that hospitals really are corporations by arguing that this is not relevant to evaluations of the appropriateness of the corporate model. PMID- 11055037 TI - The decision making process regarding the withdrawal or withholding of potential life-saving treatments in a children's hospital. AB - OBJECTIVES: To investigate the factors considered by staff, and the practicalities involved in the decision making process regarding the withdrawal or withholding of potential life-sustaining treatment in a children's hospital. To compare our current practice with that recommended by the Royal College of Paediatrics and Child Health (RCPCH) guidelines, published in 1997. DESIGN: A prospective, observational study using self-reported questionnaires. SETTING: Tertiary paediatric hospital. PATIENTS AND PARTICIPANTS: Consecutive patients identified during a six-month period, about whom a formal discussion took place between medical staff, nursing staff and family regarding the withholding or withdrawal of potentially life-sustaining treatments. The primary physician and primary nurse involved in the discussion were identified. METHOD: Two questionnaires completed independently by the primary physician and nurse. RESULTS: Twenty-two patients were identified (median age 1 year; range 1 day-34 years). In 20 cases treatment was withdrawn or withheld, in two cases treatment was continued. Nursing staff considered family wishes and family perceptions of patient suffering as significantly more important factors in decision making than medical staff, who considered prognostic factors as most important. In only two cases were the patient's expressed wishes apparently available. In most cases staff considered the patient's best interests were served and the process would not be enhanced by the involvement of an independent ethics committee. The exceptions were those cases in which treatment was continued following disagreement between parties. CONCLUSIONS: Our current practice is consistent with that recommended by the RCPCH. The contribution of the patient, provision of staff counselling and general practitioner (GP) involvement were identified as areas for improvement. PMID- 11055038 TI - Ethical dilemmas in palliative care: a study in Taiwan. AB - OBJECTIVES: To investigate the incidence and solution of ethical dilemmas in a palliative care unit. DESIGN: Health care workers recorded daily all dilemmas in caring for each patient. SETTING: Palliative care unit of National Taiwan University Hospital in Taiwan. PATIENTS: Two hundred and forty-six consecutive patients with terminal cancer during 1997-8. MAIN MEASUREMENT: Ethical dilemmas in the questionnaire were categorised as follows: telling the truth; place of care; therapeutic strategy; hydration and nutrition; blood transfusion; alternative treatment; terminal sedation; use of medication, and others. RESULTS: The type and frequency of ethical dilemmas encountered were: place of care (33.3%); truth-telling (32.1%); hydration and nutrition (25.2%); therapeutic strategy (24.8%), and use of medication (19.1%). Ethical problems relating to the place of care and to therapeutic strategy were unlikely to be solved with increased hospital stay and some ethical dilemmas remained unsolved even in the final week in hospital, including place of care (23.2%), truth-telling (17.1%) and therapeutic strategy (11.4%). Problems of truth-telling occurred in nearly half (42.6%) of patients over sixty-five-years-old. Conflicts about blood transfusion were experienced in all patients below 18-years-old, and the dilemmas concerning the place of care occurred most frequently with head and neck cancer patients (43.8%). CONCLUSIONS: The solution of ethical dilemmas required refocusing by medical professionals on the importance of continuing communication. Improved ethical training for professionals would contribute to solving the moral dilemmas of palliative care. PMID- 11055039 TI - Personal values and cancer treatment refusal. AB - This pilot study explores the reasons patients have for refusing chemotherapy, and the ways oncologists respond to them. Our hypothesis, generated from interviews with patients and oncologists, is that an ethical approach that views a refusal as an autonomous choice, in which patients are informed about the pros and cons of treatment and have to decide by weighing them, is not sufficient. A different ethical approach is needed to deal with the various evaluations that play a role in treatment refusal. If patients forgo further treatment, while curative or palliative methods are available, there is no perspective from which to integrate the weighing of pros and cons of treatment and the preferences and values of individual cancer patients. A discrepancy thus results as regards what "good reasons" are, evoking misunderstandings or even breaking off communication. Suggestions are given for follow up research. PMID- 11055040 TI - British community pharmacists' views of physician-assisted suicide (PAS). AB - OBJECTIVES: To explore British community pharmacists' views on PAS, including professional responsibility, personal beliefs, changes in law and ethical guidance. DESIGN: Postal questionnaire. SETTING: Great Britain. SUBJECTS: A random sample of 320 registered full-time community pharmacists. RESULTS: The survey yielded a response rate of 56%. The results showed that 70% of pharmacists agreed that it was a patient's right to choose to die, with 57% and 45% agreeing that it was the patient's right to involve his/her doctor in the process and to use prescription medicines, respectively. Forty-nine per cent said that they would knowingly dispense a prescription for use in PAS were it to be legalized and 54% believed it correct to refuse to dispense such a prescription. Although 53% believed it to be their right to know when they were being involved in PAS, 28% did not. Most pharmacists (90%) said that they would wish to see the inclusion of a practice protocol for PAS in the code of ethics of the Royal Pharmaceutical Society of Great Britain (CE-RPSGB) in the event of a change in the law on PAS. In addition, 89% would wish to see PAS included in the Conscience Clause of the CE-RPSGB. Males were found to be significantly less likely to favour PAS than females (p < 0.05), as were those declaring an ethnic/religious background of consideration when dealing with ethical issues in practice compared with their counterparts (p < 0.00005). CONCLUSION: Pharmacists view their professional responsibility in PAS to be more obligatory than a physician's, in having to provide the means for PAS. It is worrying that a proportion of the respondents prefer to remain in ignorance of the true purpose of a prescription for PAS; a finding at odds with current developments within the pharmaceutical profession. A practice protocol for PAS and an extension of the conscience clause should be considered in the event of PAS becoming legal. Such measures would allow the efficient provision of the pharmaceutical service whilst at the same respecting the personal beliefs of those who object to cooperating in the ending of a life. PMID- 11055041 TI - Euthanasia--a dialogue. AB - A terminally ill man requests that his life be brought to a peaceful end by the doctor overseeing his care. The doctor, an atheist, regretfully declines. The patient, unsatisfied by the answer and increasingly desperate for relief, presses the doctor for an explanation. During the ensuing dialogue the philosophical, ethical and emotional arguments brought to bear by both the doctor and the patient are dissected. PMID- 11055042 TI - Why some Jehovah's Witnesses accept blood and conscientiously reject official Watchtower Society blood policy. AB - In their responses to Dr Osamu Muramoto (hereafter Muramoto) Watchtower Society (hereafter WTS) spokesmen David Malyon and Donald Ridley (hereafter Malyon and Ridley), deny many of the criticisms levelled against the WTS by Muramoto. In this paper I argue as a Jehovah's Witness (hereafter JW) and on behalf of the members of AJWRB that there is no biblical basis for the WTS's partial ban on blood and that this dissenting theological view should be made clear to all JW patients who reject blood on religious grounds. Such patients should be guaranteed confidentiality should they accept whole blood or components that are banned by the WTS. I argue against Malyon's and Ridley's claim that WTS policy allows freedom of conscience to individual JWs and that it is non-coercive and non-punitive in dealing with conscientious dissent and I challenge the notion that there is monolithic support of the WTS blood policy among those who identify themselves as JWs and carry the WTS "advance directive". PMID- 11055043 TI - Medical confidentiality and the protection of Jehovah's Witnesses' autonomous refusal of blood. AB - Mr Ridley of the Watch Tower Society (WTS), the controlling religious organisation of Jehovah's Witnesses (JWs), mischaracterises the issue of freedom and confidentiality in JWs' refusal of blood by confusing inconsistent organisational policies with actual Biblical proscriptions. Besides exaggeration and distortion of my writings, Ridley failed to present substantive evidence to support his assertion that no pressure exists to conform to organisational policy nor systematic monitoring which compromises medical confidentiality. In this refutation, I present proof from the WTS's literature, supported by personal testimonies of JWs, that the WTS enforces its policy of blood refusal by coercive pressure to conform and through systematic violation of medical confidentiality. Ridley's lack of candour in dealing with the plea of dissident JWs for freedom to make personal and conscientious decisions regarding blood indicates that a serious breach of ethics in the medical care of JWs continues. The medical community should be seriously concerned. PMID- 11055044 TI - Mental incapacity and restraint for treatment: present law and proposals for reform. AB - The House of Lords in F v West Berkshire Health Authority [1989] considered the lawfulness of providing care and treatment for a mentally incapacitated adult. They did not, however, directly consider the use of restraint to enable the provision of care in the face of resistance from the patient. The law has since had good cause to give consideration to this important issue. This paper establishes the present law in the context of using restraint to deliver care. Although the legal principles established have derived from what might be considered to be "hard cases", life-and-death cases, they apply to all aspects of routine medical, dental and nursing care. Further, the paper considers the recent government proposals and the effect those proposals may have on the routine care of such patients. PMID- 11055045 TI - The morality of coercion. AB - The author congratulates Dr Brian Hurwitz, who recently reported the successful "intimidation" of an elderly competent widow into accepting badly needed therapy for a huge ulcerated carcinoma. He reports approvingly of the Israeli Patients' Rights Law, enacted in 1996, which demands detailed informed consent from competent patients before permitting treatment. But the law also provides an escape clause which permits coercing a competent patient into accepting life saving therapy if an ethics committee feels that if treatment is imposed the patient will give his/her consent retroactively. He suggests this approach as an appropriate middle road between overbearing paternalism and untrammelled autonomy. PMID- 11055046 TI - Futility has no utility in resuscitation medicine. AB - "Futility" is a word which means the absence of benefit. It has been used to describe an absence of utility in resuscitation endeavours but it fails to do this. Futility does not consider the harms of resuscitation and we should consider the balance of benefit and harm that results from our resuscitation endeavours. If a resuscitation is futile then any harm that ensues will bring about an unfavourable benefit/harm balance. However, even if the endeavour is not futile, by any definition, the benefit/harm balance may still be unfavourable if the harms that ensue are great. It is unlikely that we will ever achieve a consensus definition of futility and certainly not one that is applicable to every patient undergoing resuscitation. In the meantime our use of the term "futile", in the mistaken belief that it tells us whether it is worth resuscitating or not, has no utility as it will never succeed in telling us this. Furthermore we risk causing offence by use of the term and we risk harming the patient's autonomy by using futility as an overriding force. Instead we should consider the utility of our endeavours, for which an assessment of the harms of resuscitation should be added to our considerations of its benefit. This balance of benefit and harm should then be evaluated as best it can be from the patient's perspective. The words futile and futility should be abandoned by resuscitationists. PMID- 11055047 TI - Parental duties and untreatable genetic conditions. AB - This paper considers parental duties of beneficence and non-maleficence to use prenatal genetic testing for non-treatable conditions. It is proposed that this can be a duty only if the testing is essential to protect the interests of the child i.e. only if there is a risk of the child being born to a life worse than non-existence. It is argued here that non-existence can be rationally preferred to a severely impaired life. Uncontrollable pain and a lack of any opportunity to develop a continuous self are considered to be sufficient criteria for such preference. When parents are at risk of having a child whose life would be worse than non-existence, the parents have a duty to use prenatal testing and a duty to terminate an affected pregnancy. Further, such duty does not apply to any conditions where the resulting life can be considered better than non-existence. PMID- 11055048 TI - Interpretations, perspectives and intentions in surrogate motherhood. AB - In this paper we examine the questions "What does it mean to be a surrogate mother?" and "What would be an appropriate perspective for a surrogate mother to have on her pregnancy?" In response to the objection that such contracts are alienating or dehumanising since they require women to suppress their evolving perspective on their pregnancies, liberal supporters of surrogate motherhood argue that the freedom to contract includes the freedom to enter a contract to bear a child for an infertile couple. After entering the contract the surrogate may not be free to interpret her pregnancy as that of a non-surrogate mother, but there is more than one appropriate way of interpreting one's pregnancy. To restrict or ban surrogacy contracts would be to prohibit women from making other particular interpretations of their pregnancies they may wish to make, requiring them to live up to a culturally constituted image of ideal motherhood. We examine three interpretations of a "surrogate pregnancy" that are implicit in the views and arguments put forward by ethicists, surrogacy agencies, and surrogate mothers themselves. We hope to show that our concern in this regard goes beyond the view that surrogacy contracts deny or suppress the natural, instinctive or conventional interpretation of pregnancy. PMID- 11055049 TI - [Factorial and discriminant analyses of neuropsychological variables in familial and sporadic late onset Alzheimer disease]. AB - INTRODUCTION: Prevalence of late onset Alzheimer's disease (LOAD) both familial and sporadic is increasing with the raising proportion of third-age population. There are evidences either supporting or rejecting the existence of differences in the behavior of neuropsychological variables between familial and sporadic cases of LOAD. OBJECTIVE: To identify neuropsychological variables discriminating between familial and sporadic cases of LOAD, in order to detect clinical manifestations that may provide information on the pathological process of the neurodegenerative process. PATIENTS AND METHODS: Using sequential sampling, we selected individuals affected by LOAD according to the criteria of the DSM-IV and NINCS-ADRDA. The following neuropsychological protocol was used: CERAD, Wisconsin, Phonological Fluency, Rey's Figure, Raven, A Cancellation Test, WAIS (Arithmetic); also used were: Global Deterioration Scale, Functional Assessment Staging of Reisberg (FAST), Barthel and Yesavage. Parametrical and non parametrical univariate, factorial (principal components) and discriminant analyses were performed. In total, 52 patients were analyzed (average age: 74.8 years; mean age at onset of the disease: 69 years; time of disease's evolution: 5.7 years; average of educational level: 6.4 years). RESULTS: No significant statistical differences were found in clinical or neuropsychological variables between familial and sporadic cases of LOAD. Additionally, neither variables nor models were detected discriminating significantly between them. CONCLUSION: Familial and sporadic cases of LOAD present the same clinical and neuropsychological phenotype which makes very probable that sporadic cases are low penetrance familial ones. PMID- 11055050 TI - [Hereditary neuropathy with liability to pressure palsies (tomaculous neuropathy). Clinical, electrophysical and molecular study of two affected families]. AB - INTRODUCTION: Hereditary neuropathy with liability to pressure palsies (HNPP) is an autosomal-dominant disorder of peripheral myelin characterized by episodes of recurrent mononeuropathies usually involving nerves at common sites of entrapment and compression. Additional features include evidence of a diffuse demyelinating sensorimotor polyneuropathy on nerve conduction studies, focal myelin thickening (tomacula) on sural nerve biopsy, and a 1.5 Mb deletion on 17p11.2 encompassing the peripheral myelin protein 22 (PMP22) gene in most families. PATIENTS AND METHODS: Two girls, aged 10 and 12 years, presented with peroneal nerve palsy and peroneal nerve palsy plus tibial nerve palsy, respectively. In none case, a clear causal factor was recognizable. Neurophysiological studies: both cases showed diffuse sensory and motor nerve conduction velocity slowing and prolongation of distal motor latencies. In addition, features of focal entrapment neuropathy were obtained at proximal peroneal nerve level. Sural nerve biopsy: large axons demyelination, redundant myelin and intraaxonal looping (case 1) and tomaculas (case 2). Molecular genetics: 17p12-p11 deletion was demonstrated in both affected girls, the mother and two maternal uncles of case 1 and in the father, paternal grandfather and two paternal uncles of case 2. CONCLUSIONS: NHPP should be suspected in cases of peripheral neuropathy without clearly recognizable cause. Electrophysiological and molecular studies permit both delineation of the condition and identification of otherwise clinically normal family members. PMID- 11055051 TI - [Modifications of sympathetic tone induced by acupuncture reflex. Sympathetic electrical response and stimulus of 6PC]. AB - INTRODUCTION: From certain experimental data obtained by experimental myocardial perfusion and clinical and animal studies, it would seem that this corroborates the finding that puncture of the acupuncture point (Neiguan, 6PC) has an effect on organic vegetative tone. OBJECTIVE: We aimed to establish, by means of a simple, conclusive, objective measuring system such as sympathetic electrical response, whether this point had this action and whether its effect was nonspecific or due to activation of a nervous tract subjacent to this point. PATIENTS AND METHODS: We assessed the sympathetic electrical response in 29 healthy volunteers in whom basal values were recorded and again after orthodox acupuncture stimulus in 6PC, and compared with data obtained of the effects caused by stimulus of an extra-acupunctural point, of a different acupunctural point of the same channel, of bipolar stimulation of the median nerve at the level of 6PC and with the modifications generated in the response by rest lying down during the period of the experiment. RESULTS: A significant increase was observed in the P1 latency of the sympathetic electrical response and significant reduction in its total amplitude, when 6PC was stimulated, an effect not observed in any of the other conditions of the experiment. CONCLUSION: We conclude that puncture stimulation of 6PC causes significant changes in sympathetic tone, which are not due to nonspecific effects of skin puncture, nor activation of the subjacent median nerve alone. PMID- 11055052 TI - [Neurological complications of patients with lymphomas]. AB - INTRODUCTION: The lymphomas are neoplasias which may affect the nervous system at any stage of development, and affect the quality of life and survival time of these patients. OBJECTIVES: To identify the neurological complications of patients with lymphomas, determine the survival time and the cause of death in these patients. PATIENTS AND METHODS: We made a prospective study in 270 patients with the diagnosis of lymphoma who were admitted to the Hospital Clinico Hermanos Ameijeiras Ciudad de la Habana (Cuba). The complications were classified as direct or indirect, the average survival time was determined according to the Kaplan-Meier curve and the cause of death was established with anatomopathological confirmation. RESULTS: We found 26 patients to have neurological complications. Of 188 patients with non-Hodgkin lymphomas, 12.2% had neurological disorders and in these patients leptomeningeal infiltration was the main neurological complication. In the 82 patients with Hodgkin's disease, 3.6% had neurological disorders of which herpes zoster infection was the commonest. The average survival time following diagnosis and the neurological features was 9.7 months. The neurological complication was the cause of death in 57.1% of those who died. CONCLUSIONS: The patients with lymphomas had direct and indirect complications, with an average survival time of less than one year, and most died of a nervous system complication. PMID- 11055053 TI - [Burden and quality of life in carers of patients with Alzheimer type dementia]. AB - OBJECTIVE: To analyze the factors which affect the burden and quality of life in carers of patients with Alzheimer's disease. MATERIAL AND METHODS: From a sample of 234 carers of patients in the Unit for Assessment of Memory and Dementia with a diagnosis of probable Alzheimer's disease, according to the scale of National Institute of Neurological and Communicative Disorders and Stroke/Alzheimer's Disease and Related Disorders Association (NINCS-ADRDA), and of minimal or slight severity, according to the Cambridge Mental Disorders of the Elderly Examination (CAMDEX) criteria, we obtained sociodemographic variables and administered the following questionnaires: Neuropsychiatric Inventory (NPI), Rapid Disability Rating Scale (RDRS-2), Quality of Life Questionnaire (QLQ) and Burden Interview (BI). RESULTS: The sex of the carers, the hours of attention to the basic activities of daily life (BADL) and the Cambridge Cognitive Examination (CAMCOQ), Minimental State Examination (MMSE), QLQ, NPI and RDRS-2 scores were related to the BI score. Multiple regression accepted the scoring for NPI, RDRS-2 and QLQ in the model. The QLQ score was associated with male sex of the patient, the age of the carer, employment status, whether or not he lived with the patient, with the family relationship, the hours of attention to the BADL and the scores on CAMCOG, MMSE, RDRS-2, NPI and BI. The multiple regression model included the age of the carer, the BI score and the hours of attention to the BADL. CONCLUSION: The non cognitive symptoms, functional disability and poor perception of quality of life are factors affecting the burden and age of the carer, the hours of attention to the BADL and the burden affecting quality of life. PMID- 11055054 TI - [Neurological sciences based on evidence]. AB - OBJECTIVE: An exhaustive search of reported metanalysis from any medical speciality is described. MATERIAL AND METHODS: Search of papers included in MEDLINE or EMBASE between 1973-1998. A descriptive analysis of the reported papers (frequency tables and graphics) is described, including differences of mean of reported metanalysis papers by medical speciality and year. RESULTS: 1,514 papers were selected and classified. Between 1977-1987 overall mean of reported studies of neurologic metanalysis (1.20 +/- 1.10) was significatively inferior to the 1988-1998 (11.20 +/- 7.85) (p < 0.001). Global number of neurologic metanalysis was positively correlated (p < 0.05) with the number of studies about fundamentals and methodology during the study period. CONCLUSIONS: A progressive increase in the number of reported neurologic metanalysis since 1977 can be demonstrated. Diffusion of knowledge about fundamentals and methodology of metanalysis seems to have drawn and increase in performing and reporting this kind of analysis. PMID- 11055055 TI - [MRI guided stereotactic thalamotomy for the treatment of the neurogenic pain]. AB - INTRODUCTION: Medial thalamotomy is one of the first stereotactic operations to have been used for neurogenic pain, has a low complication rate and no risk of the development of iatrogenic neurogenic pain. It represents selective local relief for all types of pain, without causing somatosensorial deficit. PATIENTS AND METHODS: We did 39 posteromedial thalamotomies in patients with persistent intractable pain due to various disorders. The pain was assessed pre- and postoperatively on the VAS (Visual Analogic Scale). RESULTS: Half of the patients operated on had relief of pain after thalamotomy. In 84% (n = 39) of our cases this relief occurred on the second day, in 70% (n = 35) after three months, in 63% (n = 27) after six months, in 64% (n = 25) after nine months, in 62% (n = 23) of the patients after 12 months, and in 62% (n = 22) after 24 months. Three patients had temporary complications and one a permanent complication, but this did not make him an invalid. CONCLUSION: Posteromedial stereotactic thalamotomy under MR guidance can provide safe, effective treatment for persistent, intractable pain. PMID- 11055056 TI - [Discriminant capacity of neuropsychological efficiency in the Alzheimer's disease]. PMID- 11055057 TI - [The epidemic of neuropathy in Cuba: lessons learned]. PMID- 11055058 TI - [Angioplasty for intracranial internal carotid artery stenosis]. AB - INTRODUCTION: Percutaneous transluminal angioplastia has in recent years become an alternative to surgery and increasingly used for revascularization of the extracranial arteries in patients with cerebral ischemia. However, intracranial angioplasty is a technique which is still not widely used since it is technically more difficult and until now endoprotheses (stents) have not been available specifically designed for intracranial territory. CLINICAL CASE: A 73 year old patient with extensive extracranial and intracranial atheromatous lesions, multiple vascular risk factors and cardiac ischemia which contraindicated surgical treatment which was treated consecutively by angioplasty and angioplasty with implantation of stents in both carotid bifurcations. Subsequently, he was treated by angioplasty for a stenosing lesion of 90% of the right carotid siphon with clinical and hemodynamic repercussions. Following the procedure, which was well-tolerated by the patient, there was clinical and angiographic improvement and return to normal of the hemodynamic parameters measured by transcranial Doppler. CONCLUSIONS: Angioplasty is a technique which may be used in intracranial stenosing atheromatous lesions. Even without perfect angiographic correction, adequate blood flow is established in the hemodynamically affected lesions. As far as we know this is the first case of intracranial angioplasty of a lesion of the carotid siphon reported in Spain. PMID- 11055059 TI - [Absence of pyramidal signs in pyramidal tract postischemic Wallerian degeneration]. AB - INTRODUCTION: Wallerian degeneration (WD) is the irreversible axonal and myelin damage after the injury to the proximal portion of the axon or its cell body. The most frequent cause of WD in the central nervous system is ischemic stroke. Various studies have related the presence of pyramidal tract WD with the severity of motor deficit and partial motor improves. We present a patient with pyramidal tract WD without motor sequelae. CLINICAL CASE: A 55 years old man, hypertense and heavy smoker, suffered a sudden episode of dysarthria and left hemiparesis. Routine analysis showed hypercholesterolemia and an aortic valvular sclerosis on an echocardiogram. Cranial magnetic resonance imaging (MRI) showed multiple supratentorial lacunar infarctions. He was discharged without deficits, antiaggregated with aspirin. Six months later, he suffered a sudden episode of dysarthria. A new cranial MRI disclosed WD of the right pyramidal tract without pyramidal signs on neurologic exam. CONCLUSIONS: Presence of WD on the pyramidal tract is related with pyramidal disability in diverse degree but can develop a complete motor rehabilitation. We present a case of WD of the pyramidal tract without pyramidal deficits that supports the role of supplementary motor areas on motor rehabilitation. PMID- 11055060 TI - [Headaches, otalgia and peripheral facial palsy as a form of presentation of neurosyphilis]. AB - INTRODUCTION: Decrease in incidence of neurosyphilis over the last few decades implies that clinicians consider less frequently this diagnosis. On the other hand, some reports suggest an increase in atypical forms of this disease that represent an additional reason for missing this diagnosis. CLINICAL CASE: We report on a 16 year-old immunocompetent black female from Guinea-Bissau presented with headaches, ear pain, hearing loss and peripheral facial paralysis. A cranial CT scan showed a hypodense area in the left cortico-subcortical zone and a contrast enhancement on the left pontocerebellar angle and internal auditory meatus. On the third day of admission a diagnosis of meningitis was made, with high titles of VDRL and TPHA in CSF and serum, leading to a diagnosis of neurosyphilis. The epidemiological aspects of this case suggest either a late congenital syphilis or an infection as a result of a blood transfusion administered seven years earlier in Guinea-Bissau. CONCLUSION: This rare form of presentation of neurosyphilis emphasizes the importance of considering systematically this diagnosis, even in the context of atypical presentations. PMID- 11055061 TI - [Glioblastoma multiforme in a patient with a second complete remission of Hodgkin's disease]. AB - INTRODUCTION: Treatment of Hodgkin's disease contributes to the appearance of second malignant neoplasias such as glioblastoma multiforme. Amongst the known etiological hypotheses is alteration of the immune system secondary to chemotherapy. We seek new hypotheses relating this rare association. With better understanding of the causes provoking the appearance of these tumors new strategies could be designed for treatment. CLINICAL CASE: A 26 year old man who was in a second complete remission of a nodular sclerosis type of Hodgkin's disease complained of headache and evening fever. Magnetic resonance showed a right frontal 6 x 5 x 4 cm cystic lesion with a vascular solid zone, with irregular margins and surrounded by vasogenic oedema. On investigation there were reduced levels of immunoglobulins G (522 mg/dl)) and M (38 mg/dl). The lesion was removed surgically and histological studies confirmed the diagnosis of glioblastoma multiforme. CONCLUSIONS: Although immunosuppression may contribute to the appearance of secondary neoplasia, it is not the only hypothesis in the case of glioblastoma multiforme occurring in a patient with Hodgkin's disease. In the literature similar cases, both isolated and familial, have been reported and may have been affected by genetic or environmental factors which are as yet unknown. PMID- 11055062 TI - [The neuropathy epidemic in Cuba: eight years of investigation and follow-up]. AB - INTRODUCTION: The authors describes the past eight years, since an epidemic started in Cuba which mainly affected the nervous system, particularly the optic nerve and the peripheral nerves. It is thought to have been the biggest epidemic involving the nervous system this century, although there may be over-diagnosis. The cause is controversial but is probably nutritional imbalance with additional toxic elements also in some cases. At the present time the continuous, low, notification of new cases means that it is endemic. DEVELOPMENT: In this paper we consider the clinical and electrophysiological aspects characteristic of the disorder, the degree of involvement, evolution, incidence and annual follow-up of cases from 1992 to date and their differential diagnosis. We describe the national programme of healthcare for control of the disease in Cuba, the measures designed to reduce the number of patients, most of the results of clinical and epidemiological studies and the possible causes. We also discuss the most likely physiopathological hypotheses. PMID- 11055063 TI - [Frontal lobes: the executive brain]. AB - OBJECTIVE: To describe the current basic functional neuroanatomy of the frontal lobes and complex cognitive processes associated with this wide brain zone. DEVELOPMENT: We reviewed recent studies with neurofunctional interest. We structured the frontal zones and the cognitive functions more specifically humans, named 'executive functions'. We classified the frontal syndrome into more specific syndromes; and, we reviewed the fronto-cortical and subcortical connections, which are the basis of the frontal zones and functions. CONCLUSIONS: The frontal lobe is not a single anatomical and functional brain region. Regions and fronto-cortical and subcortical circuits within the frontal lobe are associated with motor functions and cognitive processes highly specialized, which may be differently affected. PMID- 11055064 TI - [Proposals for the treatment of children with West's syndrome]. AB - OBJECTIVE: The justification of guidelines for the treatment of children with infantile spasms and West's syndrome. DEVELOPMENT: We review, by means of prospective studies of suitable methodology, the efficacy and side-effects of adrenocorticotropin, sodium valproate, vigabatrin and other drugs used in the treatment of children with West's syndrome. CONCLUSIONS: In view of the information obtained from these studies, we suggest that in children with idiopathic or cryptogenic West's syndrome, treatment should be started with oral prednisone up to a dose of 8 mg/kg/day which should be changed to vigabatrin in the case of inefficacy or side effects. In children with secondary or symptomatic West's syndrome, treatment should be started with vigabatrin, up to a maximum dose of 200 mg/kg/day and if this is ineffective or there are side-effects it should be replaced by prednisone. If neither of these forms of treatment are successful, topiramate (up to 24 mg/kg/day) or valproate (up to 200 mg/kg/day) should be given. PMID- 11055065 TI - [Education, dementia and cerebral reserve]. AB - OBJECTIVE: To make a critical evaluation of the relation between a low educational level and dementia in order to determine whether this is a real risk factor for the development of dementia. DEVELOPMENT: We review the available epidemiological evidence for and against this, paying particular attention to any possible bias which might account for the association found. As a second step we analysed, from the point of view of the cause, the relation between low educational level and dementia so as to obtain a theoretical framework for the mechanisms by which a low educational level acts. Finally, we discuss the social health relevance of this risk factor. CONCLUSIONS: A low educational level, especially illiteracy is the most important social-health risk factor for dementia, not only because of its high prevalence and degree of association, but also because of the capacity of society to manipulate it. Certain educational policies, including those concerning the adult population, may have a marked social-health effect and require study and evaluation. PMID- 11055066 TI - [Should all mentally retarded patients have cytogenetic studies done?]. PMID- 11055067 TI - [Extracranial carotid dissection presenting with cluster headache]. PMID- 11055068 TI - [Lumbosacral plexopathy and deep vein thrombosis associated with retroperitoneal malignant fibrous histiocytoma]. PMID- 11055069 TI - [Idiopathic cortical myoclonia. A case report]. PMID- 11055070 TI - [Spinal subdural haematoma complicating lumbar puncture]. PMID- 11055071 TI - [Electroretinogram and visual evoked potentials in the diagnosis of brain death]. PMID- 11055072 TI - [Breslau Neurological Institute]. PMID- 11055073 TI - A tribute to Georg Springer, MA, MD, DSc (Hon)--who laid a foundation for using the relationship of blood groups to disease as a basis for the diagnosis and treatment of patients. PMID- 11055074 TI - Blood groups and disease: a historical perspective. PMID- 11055075 TI - Blood group associations with parasites, bacteria, and viruses. AB - Although recent investigations into the human blood groups have proceeded mainly at the molecular level, the RBC remains an exquisite model to study the expression of various genes and their related proteins. Although DNA may be informative, it may not always give meaningful information regarding protein expression on cell surfaces, which is where binding occurs. Because of their easy accessibility, RBCs will continue to be used as a major tool in the investigation of the causative agents for disease, whether they be viral, bacterial, or parasitic in nature. PMID- 11055076 TI - Immunoreactive T and Tn antigens in malignancy: role in carcinoma diagnosis, prognosis, and immunotherapy. PMID- 11055077 TI - The role of blood group antigens in malignant progression, apoptosis resistance, and metastatic behavior. PMID- 11055078 TI - Erythroid cell adhesion molecules. PMID- 11055079 TI - Blood groups and diseases associated with inherited abnormalities of the red blood cell membrane. PMID- 11055080 TI - Case definitions for diseases under national surveillance. PMID- 11055081 TI - Guidelines for reporting adverse events associated with vaccine products. Division of Immunization, Bureau of Infectious Diseases, Laboratory Center for Disease Control. PMID- 11055082 TI - Canadian recommendations for the prevention and treatment of malaria among international travellers. Committee to Advise on Tropical Medicine and Travel CATMAT), Laboratory for Disease Control. PMID- 11055083 TI - Expert working group on HHV-6 and 7 laboratory diagnosis and testing. PMID- 11055084 TI - Notifiable diseases annual summary. PMID- 11055085 TI - Prostate brachytherapy--the community hospital experience. AB - One of the options for treating localized carcinoma of the prostate includes the implantation of radioactive seeds under ultrasound guidance (brachytherapy). A community hospital-based prostate brachytherapy program was started in 1992. The overall survival and disease-free survival figures for the initial 100 patients treated in this program seem comparable to patients treated by radical prostatectomy or brachytherapy in larger series. The main side effects of brachytherapy included nocturia, daytime urinary frequency, dysuria, and proctitis. These side effects were transient and decreased to less than 10% by 12 to 24 months following implantation. Prostate brachytherapy can be effectively and safely provided in a community hospital setting. PMID- 11055086 TI - Pharmacokinetic implications of HMG CoA reductase inhibitors. PMID- 11055087 TI - Confidentiality of medical records: an overview for the Connecticut physician. AB - An ongoing concern for physicians and their patients is protecting the confidentiality of medical records. The proliferation of electronic methods for collecting and storing medical information, along with the growing complexity of the health-care system, has created the need for new means to protect confidentiality. Currently, a patchwork of protections requires that physicians keep patients' medical information confidential. This article reviews the three major sources of medical confidentiality protections: Connecticut law, federal law, and professional standards, all of which provide an array of protections. A brief summary of proposed federal and state legislation is also included. The challenge of effectively maintaining confidentiality in today's electronic age has prompted the Connecticut General Assembly, Congress, and other federal agencies to consider additional safeguards. The article summarizes the relevant legal framework in order to orient medical professionals to the complexities, practice challenges, and legal aspects of maintaining confidentiality. PMID- 11055088 TI - Privacy and genetics. PMID- 11055089 TI - Malingering and symptom magnification: a case report illustrating the limitations of clinical judgement. PMID- 11055090 TI - "Much ado about something". PMID- 11055092 TI - Glomus jugulare. PMID- 11055091 TI - Universal healthcare: a bold proposal. PMID- 11055093 TI - Endoscopic view of frontal sinus polyps. PMID- 11055094 TI - Electronystagmography in Meniere's syndrome: type 3 simultaneous binaural bithermal. PMID- 11055095 TI - Vocal fold polyp, scar, and sulcus vocalis. PMID- 11055096 TI - Enhancement along the facial nerve on MRI. PMID- 11055097 TI - TICA totally implantable system for treatment of high-frequency sensorineural hearing loss. PMID- 11055098 TI - Zinc nasal gel for the treatment of common cold symptoms: a double-blind, placebo controlled trial. AB - Effective treatment for the common cold have been difficult to develop because so many different types of virus are responsible for this condition. Oral zinc has been studied as a possible means of preventing or alleviating symptoms, with mixed results. We studied a new approach to zinc therapy--an over-the-counter nasal gel formulation (Zicam)--to independently evaluate its efficacy as a treatment for the common cold. Our study was conducted at four sites over a 5 month period. The study group consisted of 213 patients with recent-onset(< or = 24) cold symptoms; 108 patients received zinc therapy, and 105 reviewed placebo. Symptom charts were used to track the duration and severity of each patient's symptoms. At study's end, the duration of symptoms was 2.3 days (+/-0.9)in the zinc group and 9.0 days (+/-2.5)in the control group--a statistically significant difference (p <0.05). These results provide evidence that zinc nasal gel is effective in shortening the duration of common cold symptoms off when taken within 24 hours of their onset. PMID- 11055099 TI - Uncontrolled central adenoid cystic carcinoma: case report. AB - Central adenoid cystic carcinomas are rare malignancies that are believed to arise in ectopic salivary gland tissue within the maxilla or mandible. We describe the diagnosis and treatment of a central adenoid cystic carcinoma in a 54-year-old man, which we believe was a recurrence of an earlier growth that had not been completely excised. We also present a review of the literature. PMID- 11055100 TI - Botulinum toxin in otolaryngology: a review of its actions and opportunities for use. AB - Botulinum toxin has several important properties that make it an ideal chemical denervator. These include its high degree of specificity for the neuromuscular junction, its ability to induce temporary and reversible denervation, and its limited degree of side effects and complications. Botulinum toxin is being used safely in a wide variety of clinical settings by many different specialists. In otolaryngologic practice, it is being administered for the treatment of at least a dozen conditions, including various dysphonias, dystonias, and spasms as well as torticollis, facial nerve paralysis, and hyperkinetic facial lines. Studies have shown that botulinum toxin injections have a high rate of success in temporarily relieving symptoms. PMID- 11055101 TI - Case report: acute management of external laryngeal trauma. AB - External laryngeal trauma is rare, accounting for less than 1% of all trauma cases seen at major centers. We report the case of a man who experienced multiple injuries, including an external laryngeal trauma. The primary signs and symptoms of his laryngeal trauma were hoarseness, hemoptysis, the loss of his laryngeal prominence (Adam's apple), neck tenderness, traumatic emphysema in the neck, and a small penetrating wound to the right of the laryngeal prominence. The patient underwent immediate tracheostomy and surgical exploration. On long-term followup, his voice quality and airway patency improved. This case illustrates the importance of rapid identification and early management of laryngotracheal trauma in a patient with multiple injuries. PMID- 11055102 TI - Intravascular papillary endothelial hyperplasia (Masson's tumor) manifesting as a lateral neck mass. AB - Intravascular papillary endothelial hyperplasia is a benign lesion of vascular origin that is caused by an excessive proliferation of endothelial cells in normal blood vessels or vascular malformations. We report the case of a 26-year old man who had such a lesion deep within the soft tissues of his neck. Imaging studies revealed a 6-cm-diameter mass, with its epicenter in the right retromandibular space. The mass extended into the right parapharyngeal space and compressed the pharynx. The mass was excised, and the patient recovered uneventfully. We discuss the management of this lesion, with emphasis on radiologic and histologic assessment and the differential diagnosis. PMID- 11055103 TI - Subperiosteal release of the floor of the mouth musculature in two cases of Pierre Robin sequence. AB - Many management approaches have been considered to relieve upper respiratory obstruction in patients with Pierre Robin sequence, but the choice of treatment is determined by the severity of the obstruction. These options include prone positioning, the use of a nasal trumpet, and surgery. One surgical technique is the subperiosteal release of the floor of the mouth musculature. The theory behind this procedure is that this musculature is under tension, and therefore it pushes the tongue upward and backward, resulting in respiratory obstruction. In theory, the release of this musculature from the mandible should alleviate the tension and hence clear the obstruction. In an attempt to objectively evaluate this theory, we performed subperiosteal release surgery on two infants. Our first patient required an emergent tracheostomy on postoperative day 2 because of the onset of surgically induced airway edema. To avoid this complication in the second patient, we performed a tracheostomy at the same time as surgery. Pre- and postoperative magnetic resonance imaging in the second patient revealed only a minimal change in the anatomy of the floor of the mouth musculature. We believe the subperiosteal release of the floor of the mouth musculature requires further evaluation before it can be considered to be effective in the surgical treatment of respiratory obstruction in Pierre Robin sequence. PMID- 11055104 TI - ["THR" vs "TRH": debate on 2 abbreviations]. PMID- 11055105 TI - [Prognostic factors associated with progression from preeclampsia to eclampsia]. PMID- 11055106 TI - [Changes in preovulatory molecular progesterone/estradiol ratio in patients treated with controlled ovarian hyperstimulation. Can they be used in the assessment of ovarian reserve?]. AB - The objective was to evaluate if the preovulatory molecular ratio between progesterone and estradiol has age-dependent changes in patients undergoing controlled ovarian hyperstimulation. There were 180 cycles of conventional in vitro fertilization which were studied. Patients were divided in three groups: Group 1 (age less than 30 years; n = 40), group 2 (age between 30 and 35 years; n = 82), and group 3 (age between 36 and 40 years; n = 58). Leuprolide acetate was used in all cases. Molecular progesterone/estradiol ratio was calculated with the following formula: [Serum progesterone (ng/mL) x 3180 (Sl x 10(3)) + serum estradiol (pg/mL) x 3.671 (Sl)]. In patients older than 38.5 years there was positive correlation between preovulatory progesterone and estradiol (R = 0.55, R2 = 0.30). There were significant difference in molecular progesterone/estradiol ratio between group 1 compared to group 2 (P less than 0.001), group 1 compared to group 3 (P less than 0.0001), as well as group 1 compared to group 2 plus group 3 (P less than 0.0001). It is concluded that molecular progesterone/estradiol ratio decreases before any endocrine evidence of ovarian aging. The value of this putative test of ovarian reserve is discussed. PMID- 11055107 TI - [Barrier family planning methods as risk factor which predisposes to preeclampsia]. AB - Preeclampsia is a pregnancy-specific syndrome, it occurs in approximately 5-10% of all pregnancies and the etiology remains unknown, but the primigravida adolescent as such as multigravid older women whom have conceived with a new sexual partner have a greater risk, this has been associated also with the use of barrier contraceptive methods that prevent exposure to sperm with the endometrial cavity. An immunological factor has been suspected because fetal antigen's could cause antigenic reaction with the maternal immunological apparatus, for first exposure at these antigens, since the fetus is considered like an allotransplantation. This is supported in some studies that report that the use of condoms, spermicides and withdrawal are associated with developing of preeclampsia in subsequent pregnancy, and another hand indicate at cohabitation preceded for long period, practiced oral sex and use of contraceptive methods that permit exposure to sperm viable with uterus decreased the prevalence of preeclampsia. To test this hypothesis, we initially used data from two groups of pregnant women, comparing the contraceptive and reproductive history of 73 pregnant women with preeclampsia and 70 pregnant women without preeclampsia. The odds ratio for preeclampsia indicated a 2.52-fold (with 95% confidence interval, 1.17 to 5.44, p < 0.05), increased risk of preeclampsia for users of barrier contraceptives compared with women using nonbarrier contraceptives methods. Other variables like socio-demographic and obstetrics analysis were not different between both groups. This study suggest that nonbarrier contraceptive methods or the exposure of paternal spermatic antigens is protective against development of preeclampsia. Further immunological studies are necessary to determine the role of contraception methods and preeclampsia. PMID- 11055108 TI - [Changes in the weight of women from rural and urban areas in normal pregnancy]. AB - Ethnographic, nutrition and genetic differences are known that determine diverse clinical outcomes all over the world and to enhanced according to geographic location. In the State of Michoacan, Mexico, the 50.7% of the population lives a rural lifestyle and the 20% are women in reproductive age with a birth rate of 37.8 x 1000 inhabitants per year, which is significant to asses weight gain behavior during pregnancy. The objective of this study was to evaluate the Gestational Weight Gain Pattern (GWGP) in the east rural area in the State of Michoacan, and comparing this with women in the urban area of Morelia city. The research was performed during six months, in which 68 women with normal pregnancy were enrolled and controlled throughout their pregnancy in the rural clinics and who received attention for labor and delivery in a hospital. The inclusion criteria was: a first consultation before the 14 weeks of gestation. Pregnancy and delivery attention was granted according with the Mexican Official Norm (Norma Official Mexicana). The results indicated that the pregnancy weight gain assessed every three months and globally were significantly different in both populations in the second and third trimesters except in the first trimester, and it was significantly higher for the urban population, p > .05. The women age at the pregnancy, the school education, the number of prenatal consultations were less in the native women, the parity was significantly higher in the rural area, p > .05. There were no differences in the Body Mass Index (BMI). PMID- 11055109 TI - [Functioning ovarian cysts in patients with and without tubal sterilization]. AB - PURPOSE OF THE STUDY: To determine by ultrasound, the frequency of function ovary cysts (FOC) in patients with surgical sterilization by bilateral tubal obstruction (BTO) compare to a similar group without previous surgery. METHOD AND MATERIALS: Between August '96 and September 97, 290 patients were divided in 2 groups of 145 each. The first group were patients with BTO and the second, without. Each group was divided in 8 equal subgroups. RESULTS: In the patients with FOC, 35 cases (24.1%) had BTO compared to 18 (12.4%) from the group without the previous surgery. In both groups a parallel increase was detected between the 36-45 years there aren't variation in the FOE curves relates to the ratio. CONCLUSIONS: The presence of FOB is significantly greater in patients with BTO than in those without this antecedent. The age factor influence by itself in a uniform way, in both groups and its assume to alterations due function declination. The ratio seem to have influence in the apparition of FOC. PMID- 11055110 TI - [Prenatal diagnosis of fetal ovarian cyst, postnatal spontaneous remission]. AB - Prenatal diagnosis of fetal ovarian cyst, spontaneously resolved in the postnatal period. INTRODUCTION: Fetal ovarian cysts are an unusual entity, which is generally discovered during an ultrasonographic fetal examination. The objective of this report is to present a clinical case of a female fetus with an ovarian cysts which was diagnose by an ultrasound (US) at 36.5 week gestation which was spontaneously resolved in the postnatal period. CLINICAL CASE: A 22 year old woman to whom an US was performed to evaluate the gestational age due to the fact that she was to be programmed for a cesarean due to vulvovaginal condyloma. The US reported a 36.5 week gestation, the fetus was female and a cyst of 50 x 44 mm diameter was found in the fetal pelvis, lateral to the bladder. The child was born two weeks later by cesarean section and without complications. The control pelvic US at one month of age showed that the cysts was 54 x 45 mm in diameter. It was decided and wait to see what would happen. At five months of age the pelvic US showed the absence of the cyst. CONCLUSION: The diagnosis of the fetal ovarian cysts was based on three ultrasonographic criteria: 1. presence of a cystic structure of regular size which is located in the lower and lateral side of the abdomen, 2. integrity of urinary and gastrointestinal tracts, and 3. female sex of the fetus. The treatment depends on the size of the cyst and the ultrasonographic images of torsion and hemorrhage. PMID- 11055111 TI - [Long-term course of liver rupture in preeclampsia. A case report]. AB - A case report is presented of one preeclamptic patient with rupture of a big right subcapsular hematoma of the liver which was misdiagnosed with cholecystitis. A cesarean delivery was performed, which resulted is stillbirth, with abruptio placentae of 50%. The emergent treatment of the liver rupture was the right hepatic artery ligation, the postoperative development was favorable. Long term follow-up using ultrasound and liver function values were performed for 14 months and showed that the lesion has regressed with a normal hepatic imaging. PMID- 11055112 TI - [Risk factors for preeclampsia. Multivariate analysis]. AB - OBJECTIVE: To determine in multivariate analysis the clinical, social, and demographic factors for preeclampsia. STUDY DESIGN: A case-control study was designed. Three hundred patients were included, divided in two groups. 150 cases with criteria diagnosis for preeclampsia. 150 patients with normal pregnancy and deliveries. The main variables analyzed were age, schooling, marital status, employment, socioeconomic status, smoking and alcohol consumption, body mass index, familiar history of preeclampsia, history of preeclampsia in previous pregnancy, parity and type of pregnancy (single or multiple). STATISTICAL ANALYSIS: For comparison of cases and controls on categorical variables, odds ratios and 95% confidence intervals were calculated, and multiple logistic regression analyses. RESULTS: Multiple logistic regression analysis showed that history of preeclampsia in previous pregnancy has OR 23.7, 95% p < 0.001, familiar history of preeclampsia OR 1.62, p < 0.08, high body mass has OR 1.60. CONCLUSIONS: The knowledge of the most important risk factors in our population could be useful for the clinical to pre-detect the patient who will develop preeclampsia. PMID- 11055113 TI - Hawaii state legislature--Act 228 medical marijuana law. PMID- 11055114 TI - The first non-heart-beating organ donor in Hawaii--medical and ethical considerations. AB - The shortage of organ donors remains a major obstacle in transplantation in Hawaii. Some patients die while waiting for a life-saving organ. Across the nation, "marginal" donors, including non-heart-beating donors are used. The authors describe the first successful non-heart-beating organ donor transplant in Hawaii, and include medical and ethical considerations. PMID- 11055115 TI - Malpractice risk assessment among different approaches for informed consent. AB - INTRODUCTION: The standards for obtaining informed consent, set forth by the Hawaii Revised Statutes, establish that it is the physician's duty to disclose what a reasonable person objectively needs to hear in order to make an informed decision. It is the purpose of this study to report the opinions of medical malpractice attorneys to survey their opinion whether full or limited disclosure of alternative treatments in informed consent is viewed as having a lower malpractice risk. METHODS: Hawaii medical malpractice attorneys viewed a compilation of arguments for and against both full and limited disclosure, and completed an opinion survey after reading samples of disclosure statements in two different case scenarios: 1) a pediatric emergency department case involving a febrile child at risk for occult bacteremia, and 2) an obstetrics case involving a woman with a postdate pregnancy. RESULTS: A vast majority of respondents believe that, in general and in the obstetrics case, full disclosure results in less liability. In the pediatrics ED case, 46% chose full disclosure as having less liability, 38% believe that the same liability exists with both full and limited disclosure, and 15% believe that limited disclosure is associated with less liability in this case. CONCLUSIONS: Hawaii attorneys with medical malpractice experience overwhelmingly agree that, in general, full disclosure is associated will less medical legal liability. Full disclosure was also the option selected as associated with less liability by a majority of attorneys in a sample obstetrical case. Opinions were more diverse in the pediatrics ED case. Many attorneys stressed that judging the risk of liability in general is difficult, and should be done on a case by case basis. PMID- 11055116 TI - Diarrhea-associated hospitalizations among children in Hawaii. AB - OBJECTIVE: To estimate the burden of diarrhea-associated hospitalizations among children in Hawaii. DESIGN: Retrospective analysis of hospital discharge records. PATIENTS: Children from 1 month through 4 years of age with a diarrhea-associated diagnosis listed on the hospital discharge record in Hawaii. SETTING: Acute care hospitals in the state of Hawaii, 1994 through 1997. RESULTS: During 1994-1997, 2288 children (11.3% of all hospitalizations for children < 5 years of age) had a diarrhea-associated diagnosis listed at hospital discharge. The average annual incidence (per 10,000 children) of diarrhea-associated hospitalizations was 62.3; the incidence was higher for children < 1 year (101.6) than for those 1 through 4 years of age (52.7). Rates for Hawaii were lower than national rates during 1993 through 1995 (US annual rate of 89 per 10,000 children). The median length of hospital stay was 2 days for diarrhea-associated hospitalizations, and 3 days for hospitalizations specifically associated with rotavirus-related diarrhea. Diarrhea-associated hospitalizations were highest during the period December through June. CONCLUSIONS: Diarrhea is an important cause of hospitalization among children in Hawaii, where approximately 1 in 32 children may be hospitalized by 5 years of age. The age-specific and temporal trends reported here are consistent with those previously described for rotavirus-associated hospitalizations, suggesting that this pathogen is a major contributor to the overall morbidity from diarrhea. PMID- 11055117 TI - [Construction of gene targeting vector for duplicating p35Nck5a gene and its application in the gene targeting of ES cells]. AB - To duplicating the regulatory subunit p35Nck5a gene of mouse neuronal cdc2-like kinase in embryonic stem (ES) cells, about 12.2 kb of pGDTV vector for p35Nck5a gene duplication was constructed. The linearized pGDTV vectors were transfected into ES cells by electroporation. 245 drug-resistant cell clones were obtained in both G418 and GANC medium and the frequency of surviving cells was 6.22 x 10(-5). The ES cell clones were identified to have duplicated the p35Nck5a gene by use of both PCR and genomic Southern blotting, and the frequency of homologous recombination is 5.08 x 10(-7). The use of negative selection gene (HSV-tk) results in 7-fold increase at selection efficiency. This study lays the foundations of preparing mouse models of Alzheimer's disease. PMID- 11055118 TI - [grp75 protects cells from injuries caused by glucose deprivation]. AB - To elucidate the functions of grp75, CHL cells over-expressing grp75 are cultured in glucose-free medium in order to simulate energy metabolic stress. Their susceptibilities to injuries caused by glucose deprivation are assessed by trypan blue exclusion, LDH leakage measurement and cytometry analysis. Data shows a stronger resistance to glucose deprivation among cells over-expressing grp75 than among cells constitutively expressing grp75. The outcome suggests that grp75 can protect cells from injuries caused by glucose deprivation. PMID- 11055119 TI - [Chemokine and chemokine receptor gene families--a phylogenetic study]. AB - In this study, the phylogenetic trees of chemokines and chemokine receptors are produced, based on distance parsimonious method, using available amino acid sequences from GenBank. The divergence of chemokine or chemokine receptors was earlier than the divergence of vertebrates. While the divergence of homologous genes from different species is in well congruent with phylogenetic relationship of those species. The molecular evolutionary rates of chemokine receptor genes are different, with CXCR4 gene scored the lowest. Chemokines and chemokine receptors originated from few ancient genes. The similarity between the virus encoded chemokines or chemokine receptors with those of host genes is a consequence of evolutionary mimicry. PMID- 11055120 TI - [Polymorphism of porcine hormone sensitive lipase gene and sequencing the partial DNA fragments]. AB - Hormone sensitive lipase (HSL) is a key enzyme in fat metabolism. The polymorphism of the pig HSL gene is studied in this paper. The SSCP (Single Strand Conformational Polymorphism) is found in the HSL gene Exon 8. It shows three genotypes (MM, MN, NN). Fat-type pig breeds of Meishan and Tongcheng pigs have more allele M, with frequencies of 0.690 M and 0.740 M, respectively; however, lean-type pig breeds of Landrace and the Large White pigs have more allele N, with frequencies of 0.847 N and 0.845 N, respectively. PCR fragments representing HSL genotypes MM and NN are sequenced. Two A-->G transitions are detected, and it resulted in corresponding changes of amino acid (Asn-->Asp and Glu-->Gly) in the HSL protein, respectively. The HSL gene can be considered as a candidate gene for fatness in pigs. PMID- 11055121 TI - [The heterozygous effect of X-ray or ENU-induced null-mutant Adh alleles on alcohol tolerance of Drosophila melanogaster]. AB - To study the mechanisms of partial dominance, 12 intragenic alcohol dehydrogenase null mutations (Adhn) in Drosophila melanogaster were used as a model system. These known sequenced mutations, either single base substitutions or small intragenic deletions (9-16 bp) were analyzed for peptide production, dimer formation and enzymatic activity of the heterodimer. Multiple mechanisms leading to partial enzyme expression in heterozygotes and a wide range of dominance varying from almost complete recessive to a high degree of dominance were found. An expression of partial dominance for alcohol tolerance was observed for all 12 Adhn under the high stress of 10% ethanol. The genetic background on which the null mutations are expressed is a major determinant for alcohol tolerance of the heterozygous adult flies. The mutations induced by X-ray had higher average dominance than that of ENU-induced mutations, and the mutations that formed nonfunctional heterodimers had the highest dominance. PMID- 11055122 TI - [Gene analysis of blast resistance in an indica variety Digu]. AB - Digu is one of the important genetic resources in rice breeding for resistance to blast disease in China. In this study, the disease resistance identification in the parental varieties and the population F1, F2 and B1F1 from the crosses involving Digu and four susceptible varieties was carried out by inoculation with two Chinese blast strains, ZB13, and ZB15. The results demonstrated that Digu had one dominant gene Pi-d(t) to ZB13, which had been reported by Li SG. The F2 populations of Digu and other ten different varieties with known resistance genes were inoculated with strains ZB13 and the results further demonstrated that blast resistance of Digu to strain ZB13 was controlled by one dominant resistance gene. PMID- 11055123 TI - [Quantitative analysis for inheritance of quality characters in indica hybrid rice]. AB - Nine quality characters of 3 sterile lines, 10 restorers and their 30 indica hybrid rice by NC II genetic design were studied. The results showed as follows: (1) No genetic segregation occurred in F2 grains in grain length, grain width, grain weight, brown rice% and head rice%, which indicated these traits are completely controlled by maternal plant genotypes. (2) Significant genetic segregation occurred in F2 grains in chalkiness score, amylose content, gelatinization temperature and gel consistency, which indicated the genetic expression of these traits is mainly controlled by endosperm genotypes. (3) The quality characters have relatively high heritabilities except brown rice % and chalkiness score. The genetic variation of quality characters was mainly caused by additive effect except grain weight. PMID- 11055124 TI - [The chromosome-specific PCR marker's screening and identification of barley 6H chromosome]. AB - Two barley 6H chromosome specific RAPD markers were obtained by screening DNA of barley Hordeum vulgare (Betzes) and wheat-barley 6H addition line with 200 primers, then the RAPD markers were changed into specific PCR markers. Checking different plant materials by the PCR markers, it revealed that there was a specific band in those materials containing 6H chromosome such as Betzes, Igri, CS6H, and there was no specific band if the material did not contain 6H chromosome, such as Triticum aestivum, Secale cereale, Agropyron intermedium, Haynaldia villosa, Thinopyrum elongatum. Therefore, those PCR markers specific to chromosome 6H of barley are established. Southern hybridization indicated that the two cloned DNA fragments belong to barley genomic specific high-copy repeat sequence and low-copy sequence in wheat and barley genomes respectively. PMID- 11055125 TI - [Physical mapping of the genes px and cld coding peroxidase and cold-regulated protein in maize (Zea mays L.)]. AB - Peroxidase plays a key role in plant disease resistance, cold stress and some developmental processes, and cold-regulated protein functions necessarily in reaction of plants on cold or heat stress. Recent studies showed that these processes in plant cells were involved in programmed cell death (PCD). Using a biotin-labelled in situ hybridization (ISH) technique, we physically mapped the genes px and cld coding peroxidase and cold-regulated protein respectively onto maize chromosomes. Both DAB and fluorescence detection systems gave the identical results, the probe uaz235 corresponding to gene px was localized onto the long arm of chromosome 2 (2L) and 7L, and csu19 corresponding to gene cld was hybridized onto 4L and 5L. The percentage distances (from the hybridization sites to centromeres) of uaz235 in 2L and 7L were 45.4 +/- 1.3 and 67.4 +/- 3.7 respectively, and those of csu19 in 4L and 5L were 68.6 +/- 2.6 and 58.2 +/- 1.6 respectively. The physical positions of px in 2L and cld in 4L coincide with those in their genetic map pattern. The results also show that both of these genes have duplicated sites in maize genome. PMID- 11055126 TI - [Comparison of genetic diversity among maize inbred lines based on RFLPs, SSRs, AFLPs and RAPDs]. AB - RFLPs, SSRs, AFLPs and RAPDs were used to detect the genetic diversity among 15 maize inbred lines. A total of 56 probe enzyme combinations, 66 SSR primers, 20 RAPD primers and 9 AFLP primer combinations were identified with polymorphism among the entries, which produced 167, 201, 180 and 87 alleles respectively. SSR markers have the highest polymorphism information content (PIC, 0.47) and AFLP markers have the lowest value (0.36), while AFLP markers possess the highest assay efficiency index (Ai, 32.4). A comparison of genetic similarity matrices revealed that the estimates of correlation coefficients based on RFLPs, SSRs, AFLPs and RAPDs were significantly correlated, but the correlation of RAPD maker data with other markers was lower. These inbred lines were classified into five groups based on four molecular markers data, which are Tangsipingtou, Luda Red Cob, Lancaster, Reid, and PN group. They are consistent with the grouping based on the available pedigree data. Based on the results, we recommend RFLPs and SSRs for genetic diversity analysis among maize germplasm. PMID- 11055127 TI - [Genetic characterization of subcbnI genes of Chlamydomonas reinhardtii]. AB - Fourteen different revertants of Chlamydomonas reinhardtii recovered with ability of biosynthesis chlorophyll b were hybridized with wile-type strain, and tetrad analysis with random sampling was performed. It appeared that sub genes resulting in cbnI gene to reverse mutation, and localize on the first chromosome. According to its linkage that differences, 5 strains carrying various mutant alleles of suppressor genes were determined. Forward hybridological analysis demonstrated that the sub genes were absent of allelic specificity and had a single genic character in response to suppression. Phenotypic analysis of the sub/Sub diplontic hybrid have verified the dominant character of mutant sub genes. The phenomenon of present various allelic sub genes and all its characters revealed that the possibility of several ways or various regulatory means exists in biosynthesis of chlorophyll b. PMID- 11055128 TI - [Improvement of nitrogen fixation efficiency and plasmid stability in Bradyrhizobium japonicum by the introduction of dctABD and parCBA/DE genes]. AB - A recombinant plasmid pHN207 containing C4-dicarboxylic acid transport genes (dctABD) from Sinorhizobium meliloti, parCBA/DE genes from pTR102 and reporter genes luxAB from pDB30 was constructed by using pLAFR3 as the vector. The pHN207 was then introduced into the Bradyrhizobium japonicum TA11 and CB1809 by bi parental mating. It was confirmed that parCBA/DE genes could increase the stability of pLAFR3 in the transconjugants under both free-living and symbiotic condition. The results of plant pot experiment indicated that the introduction of dctABD genes could significantly improve the symbiotic nitrogen fixation efficiency of TA11 and CB1809 with soybean varieties of Heilong 33, Ningzhen No. 1 and Yudou No. 1. Compared with the control, the shoot dry weight (biomass) and total nitrogen content of the plants tested were significantly increased. PMID- 11055129 TI - A prospective randomized study of 1- and 2-stage sinus inlay bone grafts: 1-year follow-up. AB - The purpose of the present study was to compare the success of and surgical differences between 1- and 2-stage sinus inlay bone grafts and implants after 1 year in function. The individual risk for implant failure in grafted areas among 1-stage patients was about twice the risk in 2-stage patients (odds ratio 2.3, CI 0.6; 8.5). The risk for implant failure in non-grafted areas was significantly lower (P < .05) than in grafted areas, regardless of the technique used. Forty edentulous patients, selected according to strict inclusion criteria from consecutive referrals, were allocated to one or other of the 2 sinus-inlay procedures. Twenty patients received bone blocks fixed by implants to the residual alveolar crest in a 1-stage procedure (group 1). In another 20 patients, particulated bone was condensed against the antral floor and left to heal for 6 months before implants were placed (group 2). An almost equal number of implants was placed in the patients of each group, 76 in the 1-stage procedure and 74 in the 2-stage procedure. Additionally, 72 and 66 implants were placed in the anterior non-grafted regions of group 1 and group 2 patients, respectively. After 1 year in function, a total of 20 implants failed in 1-stage patients, versus 11 in 2-stage patients. Sixteen and 8 implants, respectively, of these were placed in grafted bone. All but one 1-stage patient received the planned fixed prosthetic restorations, but 1 restoration was redesigned after the first year in function because of a functionally unacceptable prosthetic design. At the 1-year follow-up, one 2-stage patient lost her prosthesis as the result of multiple implant failures. Bruxism and postoperative infections were the only parameters that could be related to implant failure, however, depending on the statistical method used. PMID- 11055130 TI - The longitudinal clinical effectiveness of ITI solid-screw implants in partially edentulous patients: a 5-year follow-up report. AB - A total of 114 ITI solid-screw implants was consecutively placed in 55 partially edentulous patients and restored with 68 fixed prostheses. The patients were followed for at least 5 years in a prospective study that focused on implant success and longitudinal reactions of the peri-implant hard and soft tissues. During the study period, 5 implants failed and 15 implants were lost to follow up, resulting in a cumulative survival rate of 95.3% after 5 years of loading. The success analysis included additional strictly defined events ("first occurrence of marginal bone loss > or = 4 mm," "first occurrence of pocket depth > or = 4 mm," and "first occurrence of crevicular fluid volume > or = 2.5 mm") and resulted in a cumulative 5-year success rate of 89.0%. Median loss of marginal bone, as observed on radiographs, was 0.7 mm between implant placement and prosthetic treatment and 0.5 mm between prosthesis placement and the 5-year evaluation. Compared to the previous year's value, the annual increase in marginal bone loss did not reach a level of statistical significance between 1 and 5 years of function, so that a steady state prevailed. The incidence of lingual-palatal surfaces affected with remarkable plaque deposits increased from 13% after prosthesis placement to 23% after 5 years. Sulcus Bleeding index, probing depth, attachment level, and crevicular fluid volume were used to describe the health of the peri-implant soft tissues. The research parameters remained almost unchanged and indicated a soft tissue response within physiologic levels. Most mechanical complications were experienced during the first year of loading and were related to loosening of occlusal screws, which occurred in 8 (12%) of 68 restorations. PMID- 11055131 TI - Branemark system implants in the posterior maxilla: clinical study of 660 implants followed for 5 to 12 years. AB - Originally, osseointegrated implants were used principally in the anterior region of the mandible and maxilla, but use in the posterior segments of both arches is common today. The long-term success of implants placed in the posterior region, an environment of stronger forces and poorer bone quality, has not been thoroughly reviewed. The purpose of the present study was to review a large series of Branemark System implants placed in posterior maxillae (660 implants in 202 patients) that have been restored with fixed partial ceramometal restorations and followed for as long as 12 years after loading. Thirteen of the implants (2%) failed between placement and loading, 12 implants were lost between loading and the end of the first year, and 10 failed thereafter, 2 as the result of fractures at 3 and 4 years. The cumulative success rate is therefore 94.4% at 5 to 6 years and 93.4% after 10 years. The quality and quantity of bone appeared to have little influence on the success rate. Surgical techniques are particularly important to the success of osseointegrated implants placed in the posterior maxilla. With careful surgical planning and execution, a success rate of approximately 95% at 5 years can be achieved. PMID- 11055132 TI - Variables that influence the relationship between osseointegration and bone adjacent to an implant. AB - It is often assumed that there is a direct relationship between the bone density adjacent to an implant, as revealed by radiographs, and the percent histologic osseointegration. Moreover, the lack of standardized methods for evaluation of histologic preparations makes it difficult to compare published studies, especially as little is known about the variables that influence these measurements. In this animal study, computer-assisted lineal analysis was used to evaluate the effects of subject, tooth position, and implant surface site on measured bone density and osseointegration in a bone augmentation experiment. Three sites--coronal lingual, apical lingual, and apical facial--were analyzed around each of 6 (3.75 x 8 mm) threaded machined titanium implants, as well as the apical facial site of 21 other implants placed in the mandibular premolar area of 5 dogs. In all sites, a progressive decrease in bone density was observed from bone adjacent to the implant to that at the titanium implant surface. There was an animal effect on osseointegration, but there were no differences between the mandibular premolar locations (second, third, and fourth). Most importantly, there were significant measurable effects attributable to the surface site examined. The need for carefully standardized histologic evaluations is established. PMID- 11055133 TI - Analysis of incidence and associated factors with fractured implants: a retrospective study. AB - Osseointegrated threaded titanium screw-type implants rarely lose integration after the first year of clinical function. Implant failure can occur for other reasons, with implant fracture being one of the major reasons for late failure. The purpose of the present study was to determine the incidence of implant fracture in completely edentulous and partially edentulous arches and to determine what factors may predispose an implant to a higher fracture risk. A retrospective evaluation of 4,937 implants was performed to determine the incidence of and factors common to fractured implants from a sample of implants placed and restored in one institutional setting. Based on the results of this study, the following observations were made: implants fracture at similar rates in the maxilla as in the mandible (0.6%), implant fractures occur more frequently in partially edentulous restorations (1.5%) than in restorations of completely edentulous arches (0.2%), all observed fractures occurred with commercially pure 3.75-mm-diameter threaded implants, and prosthetic or abutment screw loosening preceded implant fracture for the majority of the implants. More studies would be helpful to further explore the relationship and progression of factors associated with implant fracture. PMID- 11055134 TI - Removal torque and histomorphometric investigation of 4 different titanium surfaces: an experimental study in the rabbit tibia. AB - This study presents a histomorphometric and biomechanical comparison of bone response to commercially pure titanium screws with 4 different types of surface topographies placed in the tibial metaphysis of 12 rabbits. Each rabbit had 4 implants placed, 2 in each tibia. The 4 surface topographies were a machined surface, a grit-blasted surface, a plasma-sprayed surface, and an acid-etched (Osseotite) surface. After a healing period of 5 weeks, histomorphometric and removal torque data revealed a significantly higher percentage of bone-to-implant contact and removal torque for acid-etched implants compared to machined, blasted, and plasma-sprayed implants. Within the limits of this short-term experimental study, the results indicated that micro-rough titanium surfaces obtained with acid-etching procedures achieved a 33% greater bone-to-implant contact over machined titanium surfaces with an abutment-type roughness and provided enhanced mechanical interlocking. PMID- 11055135 TI - Implant materials, designs, and surface topographies: their effect on osseointegration. A literature review. AB - The aim of this article was to review the literature on materials, designs, and surface topographies of endosseous dental implants. The different categories of dental implants and the parameters of their design were analyzed in relation to their effect and significance in the process of osseointegration. The events that immediately follow implantation were described, emphasizing the factors that play a role in the development of the bone-implant interface. In addition, the methods and techniques that allow qualitative and quantitative evaluation of the interfacial zone were reviewed and their clinical correlation was assessed. PMID- 11055136 TI - A multicenter report on 1,022 consecutively placed ITI implants: a 7-year longitudinal study. AB - The aim of this multicenter study was to evaluate cumulative success and survival rates of ITI implants after 7 years. A complete medical report was obtained for all 440 patients enrolled in this investigation, which involved 10 different private practices. The 1,022 consecutively placed implants were distributed between completely edentulous, partially edentulous, and single-tooth replacement cases. During the annual follow-up visit, each implant was examined both clinically and radiographically using predefined success criteria. The cumulative survival and success rates were calculated for all implants. Implant subgroups were defined according to the medical history of the patients or pooled according to various indications, locations, implant designs, or implant lengths. In each subgroup, the related cumulative success rate was statistically compared to the global cumulative success rate. Fifteen implants (1.4%) were regarded as early failures, and at the end of the follow-up, the global failure rate reached 6.6%; 30 implants (3%) were lost to follow-up. At 5 years, the cumulative survival rate was 95.4%; this declined to 92.2% at 7 years. The weakest success rates were observed for implants placed in older patients, periodontally treated patients, and completely edentulous arches. Conversely, cumulative success rates that were significantly above average were observed for patients between 40 and 60 years old without pathology, implants placed after bone regeneration, solid-screw implants, implants placed in edentulous spaces, and implants placed as single tooth replacements. This investigation has demonstrated that in these 10 private practice settings, the success rate for ITI implants remained high for up to 5 years and declined slightly between 5 and 7 years. It should be noted that in later year intervals, a relatively small number of implants remained for the analysis of cumulative success rates. PMID- 11055137 TI - Vertical alveolar ridge distraction with prosthetic treatable distractors: a clinical investigation. AB - Alveolar ridge distraction is a recent and promising technique for ridge augmentation. Since 1997, a new distraction system incorporating a distraction implant has been in use. It can be used for alveolar ridge distraction and is not removed from the alveolar ridge. Upon completion of the distraction, it remains in the alveolar process for later prosthetic treatment. Thirty-five patients were treated with distraction implants for the correction of alveolar ridge deficiency. In 10 patients with atrophy of the mandible or maxilla, 16 patients with severe defects of the alveolar process after trauma, and 9 patients with localized alveolar ridge defects after single tooth loss, alveolar ridge distraction was carried out with the aid of 62 distraction implants. The distraction implants were loaded by prosthetic superstructures 4 to 6 months after distraction. A clinical and radiologic follow-up was carried out. Periotest values were examined, and peri-implant bleeding and probing depth were registered prior to prosthetic treatment and 3, 6, and 9 months after implant loading. In 29 patients, distraction was carried out without complications or problems. Two distraction implants were lost. In 2 patients distraction was discontinued because of ankylosis of the distraction segment. In 1 patient the alveolar ridge was overcorrected, and another patient experienced a persisting hypoesthesia of the lip. For 5% of the implants, pathologic probing depth of more than 3 mm and sulcus bleeding were registered prior to prosthetic treatment. These observations decreased during the next 9 months. Periotest values were normal before the start of prosthetic treatment. There was a decrease in the Periotest values, thus an increase in implant stability, during the following 9 months. It was concluded that alveolar ridge distraction using distraction implants can be a successful technique for alveolar ridge augmentation with a low rate of complication. Acceptable esthetic and functional results can be achieved by this atraumatic technique of surgery and distraction. PMID- 11055138 TI - Development of gingival esthetics in the edentulous patient with immediately loaded, single-stage, implant-supported fixed prostheses: a clinical report. AB - Many clinicians have reported on the success of immediately loaded implants supporting a bilaterally stabilized provisional fixed prosthesis. This protocol offers several advantages, including increased masticatory function, minimized uncontrolled transmucosal loading through cross-arch stabilization, improvement of psychologic well-being, and reduction in treatment time. However, the development and maintenance of proper dentogingival esthetics in the edentulous maxilla presents substantial challenges for the implant team. This article presents the specific pretreatment diagnostic requirements for immediate loading of single-stage implants and demonstrates a new surgical technique, followed by appropriate prosthodontic management, to develop an optimal gingival profile with interdental papillae surrounding a natural-looking dentition. One hundred fifty one ITI implants were placed into 22 dental arches and immediately loaded with a 1-piece fixed prosthesis. The results of this technique over the last 5 years are presented. PMID- 11055139 TI - Placement of posterior mandibular and maxillary implants in patients with severe bone deficiency: a clinical report of procedure. AB - The purpose of this investigation was to modify the method for implant placement in the posterior parts of the arches for fixed implant-supported prostheses using minimally invasive surgery. Eighty-six implants were placed posterior to the mental foramina in patients with severely resorbed mandibles, and 75 implants were placed in the posterior severely resorbed maxilla. Bone grafting from the mandible to the maxillary sinus was performed in 9 patients with severely atrophic maxillae. In all patients, optimal use of the anatomic features of the arch was achieved by tilting the implants. Patients were followed up for 12 to 123 months after prosthesis connection (mean 18 months). Three maxillary implants were lost at the time of abutment connection: 1 in the pterygoid plate, 1 close to the posterior sinus wall, and 1 placed in the palatal cortex. One implant was mobile approximately 1 year later, apparently because of an ill-fitting prosthesis. In the mandible, no implants were lost. The method described for the treatment of edentulous arches represents an alternative therapy to several others currently in use. This minimally invasive surgical procedure should be applicable in an outpatient clinic for treatment of severely resorbed posterior portions of the arches with implant-supported prostheses. PMID- 11055140 TI - A distraction abutment system for 3-dimensional distraction osteogenesis of the alveolar process: technical note. AB - To date, distraction osteogenesis has been carried out exclusively with devices that allow distraction in one given direction only. However, the new distraction abutment system described in this article allows distraction in any functionally or esthetically desired direction following osseointegration of 1 or several implants, provided that there are adjacent teeth or other osseointegrated implants. With this abutment system, an implant fixed in a position dictated by available bone volume can be moved into a prosthetically desirable position following segmental osteotomy. Accordingly, it also allows correction of the position of implants that were placed at an early age but whose position has changed as the result of jaw growth. Compared with conventional augmentation techniques carried out before or after implant placement, this method should lead not only to a shorter overall treatment time, but also to reduced strain on the patient and better long-term prognosis for success of implants. PMID- 11055141 TI - Empowerment and continuous improvement in the United States, Mexico, Poland, and India: predicting fit on the basis of the dimensions of power distance and individualism. AB - Although variations in national cultures predominate as explanation for the belief that universal approaches to management do not exist, there have been few reports of systematic studies. Data from employees of a single firm with operations in the United States, Mexico, Poland, and India were used to test the fit of empowerment and continuous improvement practices with national culture. Using the theoretical constructs of individualism-collectivism and power distance, the authors predicted that the practices would be more congruent in some cultures than in others and that value congruence would result in job satisfaction. Using structural equations modeling, the authors found that empowerment was negatively associated with satisfaction in India but positively associated in the other 3 samples. Continuous improvement was positively associated with satisfaction in all samples. Substantive, theoretical, and methodological implications are discussed. PMID- 11055142 TI - Investigating contingencies: an examination of the impact of span of supervision and upward controllingness on leader-member exchange using traditional and multivariate within- and between-entities analysis. AB - Using a sample of 150 bank employees, span of supervision and subordinate use of influence tactics were examined as moderators of relationships between leader member exchange (LMX) and subordinate performance and organizational commitment. Raw score analyses indicate that moderators are present. Span of supervision and upward controlling influence tactics moderate relationships between LMX and both performance and commitment. However, using multiple relationship analysis and multivariate within- and between-entities analysis to assess the level of analysis results in some discrepant findings. These discrepancies are discussed, as are implications for future LMX research. PMID- 11055143 TI - Toward an integrative theory of training motivation: a meta-analytic path analysis of 20 years of research. AB - This article meta-analytically summarizes the literature on training motivation, its antecedents, and its relationships with training outcomes such as declarative knowledge, skill acquisition, and transfer. Significant predictors of training motivation and outcomes included individual characteristics (e.g., locus of control, conscientiousness, anxiety, age, cognitive ability, self-efficacy, valence, job involvement) and situational characteristics (e.g., climate). Moreover, training motivation explained incremental variance in training outcomes beyond the effects of cognitive ability. Meta-analytic path analyses further showed that the effects of personality, climate, and age on training outcomes were only partially mediated by self-efficacy, valence, and job involvement. These findings are discussed in terms of their practical significance and their implications for an integrative theory of training motivation. PMID- 11055144 TI - Performance appraisal reactions: measurement, modeling, and method bias. AB - In this study, the authors attempted to comprehensively examine the measurement of performance appraisal reactions. They first investigated how well the reaction scales, representative of those used in the field, measured their substantive constructs. A confirmatory factor analysis indicated that these scales did a favorable job of measuring appraisal reactions, with a few concerns. The authors also found that the data fit a higher order appraisal reactions model. In contrast, a nested model where the reaction constructs were operationalized as one general factor did not adequately fit the data. Finally, the authors tested the notion that self-report data are affectively driven for the specific case of appraisal reactions, using the techniques delineated by L. J. Williams, M. B. Gavin, and M. L. Williams (1996). Results indicated that neither positive nor negative affect presented method biases in the reaction measures, at either the measurement or construct levels. PMID- 11055145 TI - Goal orientation and task demand effects on motivation, affect, and performance. AB - Two studies tested the joint effects of goal orientation and task demands on motivation, affect, and performance, examining different factors affecting task demands. In Study 1 (N = 199), task difficulty was found to moderate the effect of goal orientation on performance and affect (i.e., satisfaction with performance). In Study 2 (N = 189), task consistency was found to moderate the effect of goal orientation on self-efficacy and intrinsic motivation. Results are discussed in relation to self-regulatory processes cued by goal orientations, attentional resource demands, and the need to match goal orientations to the nature of the task. PMID- 11055146 TI - Development and examination of an expectancy-based measure of test-taking motivation. AB - A 10-item multidimensional measure of test-taking motivation based on expectancy theory, the Valence, Instrumentality, Expectancy Motivation Scale (VIEMS), was developed using a student sample (N = 90) and tested using 2 samples of job applicants in a field setting (N = 296; N = 246). In Field Study 1, the VIEMS was related to test performance. Hierarchical regression analysis showed that the VIEMS explained variance in test score beyond a general measure of test motivation. In a second longitudinal field study, pretest and posttest perceptions of motivation were compared. Results indicated that expectancy was related to actual test performance, and perceived test performance accounted for variance in posttest reports of motivation after controlling for pretest levels of motivation. Test-taking motivation did not account for variance in test performance differences between African Americans and Whites in either field study. PMID- 11055147 TI - Five-factor model of personality and transformational leadership. AB - This study linked traits from the 5-factor model of personality (the Big 5) to transformational leadership behavior. Neuroticism, Extraversion, Openness to Experience, and Agreeableness were hypothesized to predict transformational leadership. Results based on 14 samples of leaders from over 200 organizations revealed that Extraversion and Agreeableness positively predicted transformational leadership; Openness to Experience was positively correlated with transformational leadership, but its effect disappeared once the influence of the other traits was controlled. Neuroticism and Conscientiousness were unrelated to transformational leadership. Results further indicated that specific facets of the Big 5 traits predicted transformational leadership less well than the general constructs. Finally, transformational leadership behavior predicted a number of outcomes reflecting leader effectiveness, controlling for the effect of transactional leadership. PMID- 11055148 TI - Test of the cross-cultural generalizability of a model of sexual harassment. AB - Sexual harassment research has been primarily limited to examination of the phenomena in U.S. organizations; attempts to explore the generalizability of constructs and theoretical models across cultures are rare. This study examined (a) the measurement equivalence of survey scales in U.S. and Turkish samples using mean and covariance structure analysis and (b) the generalizability of the L. F. Fitzgerald, F. Drasgow, C. L. Hulin, M. J. Gelfand, and V. J. Magley (1997) model of sexual harassment to the Turkish context using structural equations modeling. Analyses used questionnaire data from 336 Turkish women and 455 women from the United States. The results indicate that, in general, the survey scales demonstrate measurement equivalence and the pattern of relationships in the Fitzgerald et al. model generalizes to the Turkish culture. These results support the usefulness of the model for explaining sexual harassment experiences in a variety of organizational and cultural contexts. PMID- 11055149 TI - Stress and open-office noise. AB - Forty female clerical workers were randomly assigned to a control condition or to 3-hr exposure to low-intensity noise designed to simulate typical open-office noise levels. The simulated open-office noise elevated workers' urinary epinephrine levels, but not their norepinephrine or cortisol levels, and it produced behavioral aftereffects (fewer attempts at unsolvable puzzles) indicative of motivational deficits. Participants were also less likely to make ergonomic, postural adjustments in their computer work station while working under noisy, relative to quiet, conditions. Postural invariance is a risk factor for musculoskeletal disorder. Although participants in the noise condition perceived their work setting as significantly noisier than those working under quiet conditions did, the groups did not differ in perceived stress. Potential health consequences of long-term exposure to low-intensity office noise are discussed. PMID- 11055150 TI - Forming, changing, and acting on attitude toward affirmative action programs in employment: a theory-driven approach. AB - A model of attitude toward affirmative action programs (AAPs) was applied in 4 studies involving 1,622 participants. In Study 1, attributes people tacitly associate with AAPs were identified by open-ended elicitation. Using those attributes, an instrument was developed and administered in Studies 2, 3, and 4. In those studies, a multiplicative composite of beliefs and evaluations about the AAP attributes predicted AAP attitude, consistent with M. Fishbein and I. Ajzen's (1975) theory of reasoned action. Demographic effects on AAP attitude were partially mediated by this composite. In Studies 3 and 4, an experimental manipulation of AAP information was successful in changing AAP attitude, but in a way that polarized existing demographic differences. Study 4 also showed that AAP attitude and subjective norm jointly and uniquely predicted intentions to perform AAP-related behaviors. Intentions predicted the actual behavior of mailing postcards to political representatives reflecting participants' support for AAPs. PMID- 11055151 TI - A meta-analytic review of occupational commitment: relations with person- and work-related variables. AB - Relations between occupational commitment (OC) and several person- and work related variables were examined meta-analytically (76 samples; across analyses, Ns ranged 746-15,774). Major findings are as follows. First, OC was positively related to job-focused constructs such as job involvement and satisfaction, suggesting that attitudes toward the job itself may be a central concern in committing to one's occupation. Second, consistent with previous work, OC and organizational commitment were positively related. This relation was found to be moderated by the compatibility of the profession and the employing organization. Third, OC was positively related to job performance and had an indirect effect on organizational turnover intention through occupational turnover intention. This latter effect suggests that understanding of organizational turnover can be enhanced by incorporating occupation-related variables into turnover models. PMID- 11055152 TI - Variance in faking across noncognitive measures. AB - There are discrepant findings in the literature regarding the effects of applicant faking on the validity of noncognitive measures. One explanation for these mixed results may be the failure of some studies to consider individual differences in faking. This study demonstrates that there is considerable variance across individuals in the extent of faking 3 types of noncognitive measures (i.e., personality test, biodata inventory, and integrity test). Participants completed measures honestly and with instructions to fake. Results indicated some measures were more difficult to fake than others. The authors found that integrity, conscientiousness, and neuroticism were related to faking. In addition, individuals faked fairly consistently across the measures. Implications of these results and a model of faking that includes factors that may influence faking behavior are provided. PMID- 11055153 TI - Instrumental values of organizational citizenship behavior for promotion: a field quasi-experiment. AB - The present study examined the relationship between promotion, perceived instrumentality of organizational citizenship behavior (OCB) for promotion, and employees' OCB before and after promotion. A field quasi-experiment involving 293 tellers of a multinational bank was conducted. Both supervisors and employees provided OCB ratings 3 months before and 3 months after the promotion decision was announced. The authors found employees who perceived OCB as instrumental to their promotion and who were promoted were more likely to decline in their OCB after the promotion. PMID- 11055154 TI - In need of treatment? Merger control, pharmaceutical innovation, and consumer welfare. PMID- 11055155 TI - 1999 LeTourneau Award. An analysis of anti-kickback and self-referral law in modern health care. PMID- 11055156 TI - Attorneys caught in the web of Medicare/Medicaid fraud. An overview of an attorney's ethical duties and criminal liability in the wake of United States v. Anderson. PMID- 11055157 TI - Combustion aerosols: factors governing their size and composition and implications to human health. AB - Particulate matter (PM) emissions from stationary combustion sources burning coal, fuel oil, biomass, and waste, and PM from internal combustion (IC) engines burning gasoline and diesel, are a significant source of primary particles smaller than 2.5 microns (PM2.5) in urban areas. Combustion-generated particles are generally smaller than geologically produced dust and have unique chemical composition and morphology. The fundamental processes affecting formation of combustion PM and the emission characteristics of important applications are reviewed. Particles containing transition metals, ultrafine particles, and soot are emphasized because these types of particles have been studied extensively, and their emissions are controlled by the fuel composition and the oxidant temperature-mixing history from the flame to the stack. There is a need for better integration of the combustion, air pollution control, atmospheric chemistry, and inhalation health research communities. Epidemiology has demonstrated that susceptible individuals are being harmed by ambient PM. Particle surface area, number of ultrafine particles, bioavailable transition metals, polycyclic aromatic hydrocarbons (PAH), and other particle-bound organic compounds are suspected to be more important than particle mass in determining the effects of air pollution. Time- and size-resolved PM measurements are needed for testing mechanistic toxicological hypotheses, for characterizing the relationship between combustion operating conditions and transient emissions, and for source apportionment studies to develop air quality plans. Citations are provided to more specialized reviews, and the concluding comments make suggestions for further research. PMID- 11055158 TI - Stabilization and solidification of metal-laden wastes by compaction and magnesium phosphate-based binder. AB - Bench-scale and full-scale investigations of waste stabilization and volume reduction were conducted using spiked soil and ash wastes containing heavy metals such as Cd, Cr, Pb, Ni, and Hg. The waste streams were stabilized and solidified using chemically bonded phosphate ceramic (CBPC) binder, and then compacted by either uniaxial or harmonic press for volume reduction. The physical properties of the final waste forms were determined by measuring volume reduction, density, porosity, and compressive strength. The leachability of heavy metals in the final waste forms was determined by a toxicity characteristic leaching procedure (TCLP) test and a 90-day immersion test (ANS 16.1). The structural composition and nature of waste forms were determined by X-ray diffraction (XRD) and scanning electron microscopy (SEM), respectively. CBPC binder and compaction can achieve 80-wt% waste loading and 39-47% reduction in waste volume. Compressive strength of final waste forms ranged from 1500 to 2000 psi. TCLP testing of waste forms showed that all heavy metals except Hg passed the TCLP limits using the phosphate based binder. When Na2S was added to the binder, the waste forms also passed TCLP limits for Hg. Long-term leachability resistance of the final waste forms was achieved for all metals in both soil and ash wastes, and the leachability index was approximately 14. XRD patterns of waste forms indicated vermiculite in the ash waste was chemically incorporated into the CBPC matrix. SEM showed that waste forms are layered when compacted by uniaxial press and are homogeneous when compacted by harmonic press. PMID- 11055159 TI - Seasonal variability in CH4 emissions from a landfill in a cool, semiarid climate. AB - Methane exchange with the atmosphere was measured during three seasons at the Rooney Road landfill in Jefferson County, CO. Substantial spatial and temporal variability in exchange rates were observed. Mean fluxes to the atmosphere were 534, 1290, and 538 mg CH4/m2/day, respectively, in the fall of 1994, winter of 1994-1995, and summer of 1995. Median fluxes were 12.42, 8.62, and 5.65 mg CH4/m2/day, respectively, during those seasons. Forty-three of 177 measurements had small negative fluxes, suggesting methanotrophic activity in the landfill cover soils. Despite probable methanotrophic activity in cover soils, landfills without gas collection systems may emit substantial CH4 to the atmosphere, with large spatial and seasonal variability. PMID- 11055160 TI - A probability model for evaluating building contamination from an environmental event. AB - Asbestos dust and bioaerosol sampling data from suspected contaminated zones in buildings allowed development of an environmental data evaluation protocol based on the differences in frequency of detection of a target contaminant between zones of comparison. Under the assumption that the two test zones of comparison are similar, application of population proportion probability calculates the significance of observed differences in contaminant levels. This was used to determine whether levels of asbestos dust contamination detected after a fire were likely the result of smoke-borne contamination, or were caused by pre existing/background conditions. Bioaerosol sampling from several sites was also used to develop the population proportion probability protocol. In this case, significant differences in indoor air contamination relative to the ambient conditions were identified that were consistent with the visual observations of contamination. Implicit in this type of probability analysis is a definition of "contamination" based on significant differences in contaminant levels relative to a control zone. Detection of a suspect contaminant can be assessed as to possible sources(s) as well as the contribution made by pre-existing (i.e., background) conditions, provided the test and control zones are subjected to the same sampling and analytical methods. PMID- 11055161 TI - Biological removal of gaseous ammonia in biofilters: space travel and earth-based applications. AB - Gaseous NH3 removal was studied in laboratory-scale biofilters (14-L reactor volume) containing perlite inoculated with a nitrifying enrichment culture. These biofilters received 6 L/min of airflow with inlet NH3 concentrations of 20 or 50 ppm, and removed more than 99.99% of the NH3 for the period of operation (101, 102 days). Comparison between an active reactor and an autoclaved control indicated that NH3 removal resulted from nitrification directly, as well as from enhanced absorption resulting from acidity produced by nitrification. Spatial distribution studies (20 ppm only) after 8 days of operation showed that nearly 95% of the NH3 could be accounted for in the lower 25% of the biofilter matrix, proximate to the port of entry. Periodic analysis of the biofilter material (20 and 50 ppm) showed accumulation of the nitrification product NO3- early in the operation, but later both NO2- and NO3- accumulated. Additionally, the N-mass balance accountability dropped from near 100% early in the experiments to approximately 95 and 75% for the 20- and 50-ppm biofilters, respectively. A partial contributing factor to this drop in mass balance accountability was the production of NO and N2O, which were detected in the biofilter exhaust. PMID- 11055162 TI - Preparation of calcium silicate absorbent from iron blast furnace slag. AB - Calcium silicate hydrate (CSH) solids were prepared from hydrated lime and iron blast furnace slag in an aqueous agitated slurry at 92 degrees C. While it was hoped a minimal lime/slag ratio could be used to create near-amorphous CSH, the surface area of the product improved by increasing the lime/slag weight ratio to 2. The addition of gypsum to the lime/slag system dramatically improved the formation of surface area, creating solids with 139 m2/g after 30 hr of reaction when only a minimal amount of lime was present. The SO2 reactivity of solids prepared with gypsum greatly exceeded that of hydrated lime, achieving greater than 70-80% conversion of the alkalinity after 1 hr of reaction with SO2. The use of CaCl2 as an additive to the lime/slag system, in lieu of gypsum, also produced high-surface-area solids, 115 m2/g after 21 hr of reaction. However, the SO2 reactivity of these sorbents was relatively low given the high surface area. This emphasized that the correlation between surface area and SO2 reactivity was highly dependent on the solid phase, which was subsequently dependent on slurry composition. PMID- 11055163 TI - Distribution equilibrium of mercury (II) chloride between water and air applied to flue gas scrubbing. AB - In the literature, different values of the distribution coefficient KH for HgCl2 between water and air are present in a range that spans more than 3 orders of magnitude. In order to determine if a waste incineration scrubber solution could become saturated with regard to HgCl2, an accurate experimental determination of the distribution constant of HgCl2 at elevated temperatures is needed. In this work, the coefficient has been determined at four different temperatures between 10 and 50 degrees C. The Arrhenius expression obtained is 5.5 x 10(5) x exp[ (8060 +/- 2200)/T] with a corresponding enthalpy for the process HgCl2(aq)<==>HgCl2(g) of 67 +/- 20 kJ/mole. KH at 293 K was found to be approximately 5 x 10(-7) atm M-1, which is in almost perfect agreement with an earlier study. Applying the obtained KH values to waste incineration scrubber conditions shows that no major saturation effect will occur. PMID- 11055164 TI - Heavy metals in urban soils of East St. Louis, IL, Part I: Total concentration of heavy metals in soils. AB - The city of East St. Louis, IL, has a history of abundant industrial activities including smelters of ferrous and non-ferrous metals, a coal-fired power plant, companies that produce organic and inorganic chemicals, and petroleum refineries. A protocol for soil analysis was developed to produce sufficient information on the extent of heavy metal contamination in East St. Louis soils. Soil cores representing every borough of East St. Louis were analyzed for heavy metals--As, Cd, Cu, Cr, Hg, Ni, Pb, Sb, Sn, and Zn. The topsoil contained heavy metal concentrations as high as 12.5 ppm Cd, 14,400 ppm Cu, ppm quantities of Hg, 1860 ppm Pb, 40 ppm Sb, 1130 ppm Sn, and 10,360 ppm Zn. Concentrations of Sb, Cu, and Cd were well correlated with Zn concentrations, suggesting a similar primary industrial source. In a sandy loam soil from a vacated rail depot near the bank of the Mississippi River, the metals were evenly distributed down to a 38-cm depth. The clay soils within a half-mile downwind of the Zn smelter and Cu products company contained elevated Cd (81 ppm), Cu (340 ppm), Pb (700 ppm), and Zn (6000 ppm) and displayed a systematic drop in concentration of these metals with depth. This study demonstrates the often high concentration of heavy metals heterogeneously distributed in the soil and provides baseline data for continuing studies of heavy metal soil leachability. PMID- 11055165 TI - Heavy metals in urban soils of East St. Louis, IL. Part II: Leaching characteristics and modeling. AB - The city of East St. Louis, IL, has a history of abundant industrial activities including smelters of ferrous and non-ferrous metals, a coal-fired power plant, companies that produced organic and inorganic chemicals, and petroleum refineries. Following a gross assessment of heavy metals in the community soils (see Part I of this two-part series), leaching tests were performed on specific soils to elucidate heavy metal-associated mineral fractions and general leachability. Leaching experiments, including the Toxicity Characteristic Leaching Procedure (TLCP) and column tests, and sequential extractions, illustrated the low leachability of metals in East St. Louis soils. The column leachate results were modeled using a formulation developed for fly ash leaching. The importance of instantaneous dissolution was evident from the model. By incorporating desorption/adsorption terms into the source term, the model was adapted very well to the time-dependent heavy metal leachate concentrations. The results demonstrate the utility of a simple model to describe heavy metal leaching from contaminated soils. PMID- 11055166 TI - Inhalation transfer factors for air pollution health risk assessment. AB - To facilitate routine health risk assessments, we develop the concept of an inhalation transfer factor (ITF). The ITF is defined as the pollutant mass inhaled by an exposed individual per unit pollutant mass emitted from an air pollution source. A cumulative population inhalation transfer factor (PITF) is also defined to describe the total fraction of an emitted pollutant inhaled by all members of the exposed population. In this paper, ITFs and PITFs are calculated for outdoor releases from area, point, and line sources, indoor releases in single zone and multizone indoor environments, and releases within motor vehicles. Typical PITFs for an urban area from emissions outdoors are approximately 10(-6)-10(-3). PITFs associated with emissions in buildings or in moving vehicles are typically much higher, approximately 10(-3)-10(-1). PMID- 11055167 TI - Exposure efficiency: concept and application to perchloroethylene exposure from dry cleaners. AB - Standard approaches for computing population exposures due to specific sources of air pollutants are relatively complex. In many cases, more simple and approximate methods would be useful. This paper develops an approach, based on the concept of exposure efficiency, that may be used for estimating the impact of a source (or source class) on the integrated population exposure. The approach is illustrated by an example, which uses the concept of exposure efficiency to examine the impact of perchloroethylene emissions from dry cleaners in the United States. The paper explores the geographic variability of exposure efficiency by evaluating it for each of 100 randomly selected dry cleaners. For perchloroethylene, which has a long atmospheric residence time, the site-to-site variability in exposure efficiency is found to be relatively small. This suggests that simple exposure assessments, based on generic distributional characterizations of exposure efficiency, may be used in risk assessments without introducing appreciable uncertainty. For many compounds, like perchloroethylene, the uncertainty inherent in the estimation of cancer potency or source emissions would dominate these small errors. PMID- 11055168 TI - Calcaneal bone mineral density and mechanical strength of the metatarsals. AB - The primary aim of this study was to determine the predictive value of the bone mineral density of the calcaneus for fracture of the metatarsals. The authors report a strong positive correlation between the bone mineral density of the calcaneus and the four-point bending strength of each of the five metatarsals (r2 = 0.76, 0.64, 0.70, 0.68, and 0.78 for metatarsals 1 through 5, respectively). In addition, the relative strengths of the metatarsals and the correlation with their in vivo loads during gait as previously reported in the literature are discussed. PMID- 11055169 TI - Effect of cast and noncast foot orthoses on plantar pressure and force during normal gait. AB - A variety of plantar pressure and force measures were explored in 22 healthy individuals with excessive pronation. The measures were obtained while the subjects wore a thin-soled athletic shoe alone, a modified Root foot orthosis made from a neutral cast, and a flat noncast insole with a 6 degrees varus rearfoot post. The data obtained from subjects wearing the noncast insole differed only minimally from those obtained while they were wearing the shoe only. In contrast, the modified Root orthosis had a profound effect on foot function. Heel forces and pressures were reduced, and the rearfoot contact area was increased. Measures of force in the midfoot demonstrated substantial increases in load in this region, but the increase in area associated with the contoured device resulted in no increase in midfoot pressure measurements. Forefoot pressures were reduced both medially and laterally with the cast device in place. PMID- 11055170 TI - Dermatophyte test medium culture versus mycology laboratory analysis for suspected onychomycosis. A study of 100 cases in a geriatric population. AB - Onychomycosis is the most frequently encountered condition in podiatric practice in the United States. A variety of modalities are available to confirm the presumptive diagnosis of onychomycosis. This study was conducted to compare the results of in-office dermatophyte test medium cultures with those of mycology laboratory analysis for 100 cases of suspected onychomycosis in a geriatric population. The results demonstrated that 20% of the patients had dermatophyte involvement, 56% had saprophyte involvement, and 19% had yeast involvement. Only 50% of positive dermatophyte test medium cultures correlated with a positive microscopic fungal culture for dermatophytes. Given these results, it is questionable whether in-office dermatophyte test medium cultures should be routinely used in geriatric patients for the diagnosis of onychomycosis. The authors believe mycology laboratory testing with fluorescent potassium hydroxide preparations and microscopic fungal cultures to be superior to in-office dermatophyte test medium cultures for the diagnosis of onychomycosis in geriatric patients. PMID- 11055171 TI - Ectrodactyly-ectodermal dysplasia-clefting syndrome. AB - Ectrodactyly-ectodermal dysplasia-clefting syndrome is a rare congenital anomaly that affects tissues of mesodermal and ectodermal origin. Musculoskeletal involvement frequently requires orthopedic intervention. The authors present a review of the literature pertaining to this rare syndrome as well as a case report of a female patient who exhibited the complete clinical triad. A description of the surgical management of her condition is also presented. PMID- 11055172 TI - Ethical issues in medical journal editing. PMID- 11055173 TI - Podiatric medical resources on the Internet. A third update. PMID- 11055174 TI - Vincristine-induced neuroarthropathy (Charcot's joint) PMID- 11055175 TI - Osteoid osteoma of the distal phalanx. PMID- 11055176 TI - [Main issues of psychoneuroimmunology: Part I]. AB - Psychoneuroimmunology is concerned with interdependent links between psyche, nervous, endocrine and immune systems. The origin of this scientific discipline is a breakthrough in the way of thinking about the functioning of a person and contributes to the holistic attitude. The results of research in psychoneuroimmunology are of particular significance for psychosomatic medicine since they explain in a systemic way earlier clinical observations and scientific studies concerning the influence of stress on the health condition. Studies of the immune system conditioning are becoming a direction which is cognitively significant and clinically important. The aim of the standard immune system conditioning by pharmacological means is to activate the potential for the human body to cope with disease. PMID- 11055177 TI - [Main issues of psychoneuroimmunology: Part II]. AB - In psychoneuroimmunology links between psyche and the body are examined in the context of neurotransmitter, hormone and immuno-transmitter interaction. This allows for construction of models which show empirically verifiable links between the nervous, endocrine and immune systems. The earlier concepts of stress by Cannon and Selye focused on the physical and mental strain influence on the action of the nervous and endocrine systems. Ursin, Olff and Schedlowski introduced the concept of stress extended by an immune system reaction, which is an integral part of the alarm phase. A change in the amount of NK cells and their stress-influenced activity is an important defense mechanism of the body. It constitutes a component of the preparation for defense against potential pathogen penetration. PMID- 11055178 TI - [Psychoimmunological aspects of using the psychoactive substance by adolescents]. AB - In this research a quantitative analysis of cluster differentiation and stimulation in lymphocyte T, B, NK cells and lymphocyte T subpopulations was made, together with an evaluation of physical and psychological state of opiate dependent adolescents (Polish heroin administered intravenously). Based on the conducted research the following conclusions were drawn: the changes within the lymphocyte T composition and in their subpopulations, as well as in the NK cells indicate cell immunity disturbances in persons with a permanent pattern of opiate use (Polish heroin), not causally related to psychiatric disorders occurrence in those persons and their somatic state. The lowering of CD4 lymphocyte level in opiate-dependent persons, HIV-seronegative, can indicate an immune system defect, not causally related to HIV infection; decreased number of NK cells which are an important element of non-specific cell immunity, indicates a possibility of a greater number of infections in persons treated due to opiate dependence. PMID- 11055179 TI - [Evaluation of selected immunological parameters in alcohol dependence syndrome]. AB - In 28 patients with alcohol dependence syndrome (ADS) the study of selected immunological parameters (percentage of peripheral blood lymphocytes CD3+, CD4+, CD8+, CD19+; lymphocyte transformation without and with phytohemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM); chemiluminescence of peripheral blood granulocytes stimulated with phorbol myristate acetate (PMA)) and Instytut Merieux' skin tests (Multitest CMI) were performed. The results of immunological parameters were connected with activity of liver enzymes (aspartate aminotransferase (ASP), alanine aminotransferase (ALT), gamma-glutamyltransferase (GGT). The differences of reactivity of immune system in the tested groups of patients were observed. PMID- 11055180 TI - [Alcoholism: biology]. AB - This article discusses biochemical changes of ethyl alcohol in human organism, concentrating especially on the negative influence on the metabolism of liver. The authors describe the process of oxidation of alcohol with alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) and emphasize the role of ADH i ALDH isoenzymes in creating individual tolerance of ethanol. Two other ways of ethanol metabolism are also presented. These are: microsomal ethanol oxidation system (MEOS) connected with cytochrome P-450 and peroxisome catalase system. The article also describes the influence of alcohol and its products of metabolism on the structure of liver proteins, on different metabolic processes taking place in this organ, and on the changes in the immunological system in the course of the alcoholic liver disease. Moreover, the authors present some information about the changes in histopathological picture of liver as the result of alcohol abuse. PMID- 11055181 TI - [Suicidal attempts among young people hospitalized in the department of psychiatry of the Medical Academy in Lublin in 1990-1997]. AB - Based on clinical assessment of the case histories, the results of a retrospective suicidal research of 66 patients between the ages of 15 and 25 years who were hospitalised in Psychiatric University Hospital in Lublin between 1990 and 1997 were presented. In the paper the analysis was performed of clinical and sociodemographic aspects, significant from suicidal point of view, such as: the psychiatric diagnosis, reasons for suicidal attempts, the means of realization of the suicidal attempts, somatic conditions, alcohol and substance abuse, months of suicidal attempts, months of hospitalisations, duration of hospitalisation, repetition of suicidal acts, suicidal family history, negative interactions in family system, gender, current occupation, education, and marital status. In the examined group, six main reasons of suicidal attempts were selected, such as: 1) delusions (28% patients of the whole group), 2) hallucinatory behaviour (16% patients), 3) situational disorders (17% patients), 4) acute reactions (17% patients), 5) insight disorders (17% patients), 6) depressive disorders (11% patients). The differences between the psychotic group (with a prevalence of males) and the non-psychotic group (with a prevalence of females) proved to be significant. When taking into account the method of the suicide, there was a marked difference between the group of males (prevalence of "hard" methods, mainly hanging themselves) and the group of females (prevalence of "soft" methods, mainly drug intoxications). In the majority the families, systems of mutual interactions were disordered. The exacerbation of suicidal acts was registered in autumn-winter period. Psychoactive drug abuse was a malignant predictor of suicidal behaviour (mainly among males). PMID- 11055182 TI - [Genetic studies in autistic disorders]. AB - Since autism was first described by Leo Kanner the view on its etiology and pathogenesis has been changing. Recently there are more data on genetic and neurobiological background of autism. At the beginning it was noticed that autism appeared more frequently among boys, in population studies it was found that autism appeared more frequently among siblings, mostly among monozygotic twins. Many disorders like Tourett syndrome and tuberous sclerosis were reported in connection with autism. Recently research is focused mostly on chromosome abnormalities: chromosome 15 (locus 15q11-13), chromosome 7 (locus 7q), chromosome 16 (locus 16p) and gens of particular receptors (GABRB3, UBE3A/E6-AP, 5-HTT). These abnormalities may also be one of the causes of autism. PMID- 11055183 TI - [Pervasive developmental disorders: controversies concerning the classification of autism]. AB - Autistic Disorder was described by Leo Kanner in 1943. Since that time not only the name of this disorder (initially early infantile autism) has changed but also it's relation to other disorders. DSM-IV includes autism together with Rett's Disorder, Childhood Disintegrative Disorder, Asperger's Disorder and Pervasive Developmental Disorder Not Otherwise Specified into one category: Pervasive Developmental Disorders. The definition and contents of Pervasive Developmental Disorders raise many controversies. Differentiation between particular disorders within this category is also difficult. This paper discusses some of these problems. PMID- 11055184 TI - [About the necessity of neuropsychological assessment in the civil law]. AB - Giving expert opinions within legal psychiatry in civil cases requires more and more strict co-operation of expert psychiatrists with psychologists. Assessment of the cognitive functions with the help of neuropsychological methods is an important diagnostic element that leads to giving the right opinion. The most frequent reasons for appointing experts in psychiatry and psychology are disorders of cognitive functions as a result of various brain injuries. Dementia syndromes are particularly often subjects of doubts in preparing expert opinions as they must be distinguished from other organic dysfunctions and from the age- associated memory impairment. Considering the evidence value the most important thing is to assess all the objective data included in the medical records and subsequently to assess the testimony of the witnesses Usually people from legal circles and families of the people who make declarations of will overvalue the importance of additional examinations. Those examinations are important but they do not settle the patient's psychic state because the decisive factor is not the kind of somatic disease but the influence of that disease on the psychic state. The neuropsychologist's role in giving medical statements is going to increase together with the tendency to objectivization and qualitative assessment of intensification of respective disorders of cognitive functions when examining patients in order to give expert opinions. PMID- 11055185 TI - [An interesting example of procedural and therapeutic failure]. AB - Forty-eighth-year old woman experienced paranoid syndrome for the first time. Her twenty-seven-year old daughter, living with her, adopted pathological sensations. The patient was hospitalized by order of court. The ruling was based on the patient's husband's application and a psychiatrist's certificate. The daughter of the patient still maintained her pathological beliefs, adopted from her ill mother. What is more, similar beliefs were adopted by the second, twenty-eighth year old daughter. Hospitalization lasted only 11 days because the husband withdrew his application and, by order of court, the patient was discharged from hospital. She now lives with two psychotically induced daughters and her husband, who is under great psychical pressure in such circumstances. PMID- 11055186 TI - [On the temporary involuntary commitment]. PMID- 11055187 TI - [Citalopram (Cipramil) in monotherapy of depressed outpatients]. AB - Citalopram at a dose of 20 mg was used as monotherapy in the treatment of 91 outpatients with depression mostly of moderate intensity. The duration of treatment was 2 months. Patients were assessed by Clinical Global Impression Scale (CGI) before, after 1st and 2nd months of therapy. Twenty three patients were additionally rated after 2 weeks. Observed and spontaneously adverse events were also recorded. A large proportion of treated patients (75%) showed full or almost full remission (1 or 2 on CGI) at the end of study. There were only few mainly mild adverse events during treatment. Nausea (7.7%) and headache (3.3%) occurred most often. The author conclude that citalopram is a well-tolerated and efficacious antidepressant drug for outpatients. PMID- 11055188 TI - By any other name. PMID- 11055189 TI - Antibiotics and breastfeeding. PMID- 11055190 TI - A primary germ cell tumor of the anterior mediastinum: a case report and discussion. AB - The anterior mediastinum is the most common extragonadal location for germ cell tumors and accounts for about 50% to 70% of such neoplasms. Embryonal cell carcinomas are one of the rarest forms and account for less than 2%. We present the case of a 19-year-old, white male who was found to have a primary embryonal cell carcinoma of the anterior mediastinum. This case illustrates the subtle complaints that these patients present with, some of the problems and decisions that go into making the diagnosis, and the response to the appropriate therapy. The following discussion takes a look at the variety of germ cell tumors, the vast differential of an anterior mediastinal mass, the workup of such a mass, and the various treatments and outcomes of extragonadal germ cell tumors. PMID- 11055191 TI - The 1999 annual report of the Regional Infant and Child Mortality Review Committee. AB - The 1999 Annual Report of the Regional Infant and Child Mortality Review Committee (RICMRC) is presented. Our Regional (Minnehaha, Lincoln, and Turner Counties) incidence for Sudden Infant Death (SIDS) continues to significantly exceed the national rate. In this study, SIDS is strongly associated with prone sleeping and sleeping on soft surfaces or bedding. The Back-to-Sleep campaign that has been an important part of lowering the national SIDS rate appears to have been less successful in our region. The Regional Infant Child Mortality Review Committee therefore has elected to serve not only as a data collection committee, but has also actively engaged in community education programs directed towards providing a safer environment for our children. PMID- 11055192 TI - Regional Infant and Child Mortality Review Committee 1999 final report. PMID- 11055193 TI - [Present-day problems of hypoxia]. AB - The basic mechanism of hypoxia is energy apparatus dysfunction which is associated with ensuing inactivation of mitochondrial enzyme complexes (from the substrate to terminal portion of the respiratory chain) in oxygen deficiency, which leads to impairments of aerobic energy synthesis, energy-dependent functions, metabolism, and structure of cells. The effects of hypoxia are realized by two ways: 1) by direct impact of oxygen deficiency on the cellular bioenergy apparatus, followed by its dysfunction (bioenergetic hypoxia); 2) by indirect impact via stressor activation of the neurohumoral link that leads to the trigger of a nonspecific cascade of functional and metabolic reactions, to impaired cell oxygen supply and delivery, which also ultimately favors the development of bioenergetic hypoxia. The sequence of bioenergetic impairments is the major mechanism of any forms of hypoxia and underlies the body's individual resistance to oxygen deficiency. To correct these disorders (to restore cell energy generation processes and to maintain them at the level sufficient to perform energy-dependent functions) is the main task of antihypoxic protection. In this connection, prognostic biochemical criteria of different stages of hypoxia have been developed; a new classification of antihypoxants by the mechanism of their action is proposed by identifying a group of specific energy metabolic correctors; a combined drug therapy policy in oxygen deficiency is substantiated by taking into account the body's phenotypic features. PMID- 11055194 TI - [Lipogenesis in the brain under hypoxia]. AB - This study investigated the utilization of some radioactive precursors [2(12)C] acetate, [1-6(14)]-glucose, [5(14)C]-glutamate) for fatty acids and lipid biosynthesis in the rat brain under normal and hypoxic conditions. In severe hemic hypoxia (30-45 min after the injection of 15 mg of NaNO2/100 g body weight) there was a significant increase in 14C incorporation from glutamate into brain lipids (by 2.8 times) and into fatty acids (by 2.2 times) as compared to the control level. Enhanced lipogenesis from glutamate was demonstrated due to the activation of all alpha-ketoglutarate shunt steps. The higher lipogenesis from glutamate in the brain as a possible mechanism for this excitatory amino acid utilization under hypoxia. PMID- 11055195 TI - [Current ideas of intracellular energy maintaining mechanisms in health and disease]. AB - The paper presents the author's own findings and data available in the literature concerning the intracellular mechanisms of conversion of fatty acids to glucose and glycogen in mammalian and human tissues in health and in disease. This conversion is considered by the author to be a regular adaptive response that maintains energy homeostasis in oxygen deficiency. PMID- 11055196 TI - [Cerebral circulation in different types of brain hypoxia]. AB - The paper describes differences of hypoxic and circulatory hypoxias (i.e. brain ischemia) which cause decreases not only in the supply of O2, but in the delivery of glucose and other oxidation substrates and in venous return, which is attended by the accumulation of metabolic products in the brain tissue. It also considers the mechanisms of primary and secondary brain ischemia occurring with decreased cerebral circulation due to breakdown of cerebral blood flow autoregulation at its lower and upper borders to develop cytotoxic or vasogeneous brain tissue edema with possible compression of the microcirculatory bed in the latter case. Emphasis is laid on the significance of autoimmune reactions occurring with the impaired blood-brain barrier due to different types of cerebral circulatory disorders, which gives an insight into the cause of progressive damage to the brain in some cases despite its single damage. The paper outlines current therapies for brain ischemia, including those that exert effects on metabolic disturbances and neurosurgical reparative operations. In conclusion, the paper considers a new nontraditional way of increasing collateral CBF by decreasing blood flow pseudoturbulence with special high molecular-weight linear polymer solutions by the Thoms-effect method (1948). The prospects for using this approach in patients with brain ischemia are substantiated by a number of the established facts: 1) the above patients have higher hemodynamic blood flow resistance which may be corrected by adding a polymer solution into the sample in in vitro tests; 2) there was an inverse relationship of the intrinsic plasma concentrations of high molecular-weight fragments of DNA and hemodynamic resistance to the changes in plasma DNA properties in stroke patients. PMID- 11055197 TI - [Hypoxia and memory. Specific features of nootropic agents effects and their use]. AB - Hypoxia and hypoxic adaptation are powerful factors of controlling memory and behavior processes. Acute hypoxia exerts a differential impact on different deficits of mnestic and cognitive functions. Instrumental reflexes of active and passive avoidance, negative learning, behavior with a change in the stereotype of learning are more greatly damaged. Memory with spatial and visual differentiation and their rearrangement change to a lesser extent and conditional reflexes are not deranged. In this contract, altitude hypoxic adaptation enhances information fixation and increases the degree and duration of retention of temporary relations. Nootropic agents with an antihypoxic action exert a marked effect on hypoxia-induced cognitive and memory disorders and the magnitude of this effect depends on the ration of proper nootropic to antihypoxic components in the spectrum of the drugs' pharmacological activity. The agents that combine a prevailing antiamnestic effect and a marked and moderate antihypoxic action (mexidole, nooglutil, pyracetam, beglymin, etc.) are most effective in eliminating different hypoxia-induced cognitive and memory disorders, nootropic drugs that have a pronounced antiamnestic activity (centrophenoxine, etc.) and no antihypoxic component also restore the main types of mnestic disorders after hypoxia, but to a lesser extent. PMID- 11055198 TI - [Involvement of glutamate receptors (NMDA type) in reaction of brain neurons to anoxia of different duration]. AB - The participation of NMDA receptors in the changes caused by evoked EPSPs in the activity of calcium and phophoinositide intracellular regulatory systems (ICRS) bioelectric activity (evoked EPSPs) and in cell production of polypeptides was studied in rat olfactory cortical slices exposed to short-term anoxia (STA) and long-term anoxia (LTA). To determine NMDA receptor involvement in these functional and metabolic reactions, the slices were treated with the antagonists APV and MK-801 during anoxia of different duration (STA and LTA being 2 and 10 min, respectively) and subsequent reoxygenation. EPSPs were recorded extracellularly in response to lateral olfactory tract stimulation. The STA mediated stimulation of NMDA receptors was found to promote adaptive reactions (moderate activation of IRS, long-term potentiation of synaptic transmission) in the postanoxic period. LTA induced the long-lasting stimulation of NMDA receptors that led to the hyperactivation of calcium and phosphoinositide IRS, to the suppression of generation of EPSPs and of synthesis of polypeptides probably of adaptive action. All these events induced pathological processes in the postanoxic period. PMID- 11055199 TI - [Receptor agonists as perspective neuroprotective agents]. AB - Selective agonists of adenosine A1, GABAa, GABAb, and alpha 2-receptors (inhibitory neurotransmitter analogs) have a high neuroprotective effects (NPE) on two models of complete cerebral ischemia (CI). These NPEs are specific as they are inhibited by selective antagonists and, in addition, the selective alpha 1- and beta-receptor agonists have no NPE. The natural resistance to CI is also associated with A- and alpha 1-receptors. NPE does not result from improved cerebral blood flow. The receptor agonists protect the brain itself. They use a tolerance strategy and hypothermia is an important component, but not the only mechanism of NPE. Unlike most drugs, receptor neuroprotectors (RNP) guard not only a penumbra zone, but the core of ischemia as they are effective in complete global CI. Moreover, RNP-induced delay of irreversible lesions may increase a therapeutical window for using other drugs. RNPs are promising potential drugs in CI. PMID- 11055200 TI - [Hypoxia and nitric oxide]. AB - Cell death or survival under hypoxia is determined to a greater extent by the nature of changes in nitric oxide (NO) metabolism. Severe hypoxia causes NO overproduction that is an important factor that induces apoptotic cell death. In contrast, a cell response to moderate hypoxia enhances hypoxic resistance and restricts the mechanisms of activated apoptosis. A moderate increase in NO synthesis and storage that are characteristic of hypoxic adaptation may limit NO overproduction induced by acute hypoxia. In addition, NO-dependent mechanisms of adaptation are apparently involved in the restriction of apoptosis-specific signaling pathways. Therefore, during hypoxic adaptation, NO acts as an endogenous protective agent involved in the limitation of severe hypoxia-induced cell damages. PMID- 11055201 TI - [Efficiency of intermittent and resonance intermittent normobaric hypoxia therapy in patients with infection-dependent bronchial asthma]. PMID- 11055202 TI - [Medicine strategy in population's health care on the eve of 21-st century]. AB - The reforms have done great damage to the population's health and reduced birth rates in Russia. They were implemented as "shock therapy" which broke up a "dynamic stereotype" in the vast majority of individuals. The break-up of the "dynamic stereotype" was due to the fact that there were persons who had been grown under market economy in Russia at all. It is the circumstances that fundamentally distinguishes the conditions in Russia from those in Eastern Europe and Baltic countries. On the eve of the 21st century, the medical strategy for health care of Russia's population can be only prophylactic. This includes 3 main vectors: 1) prevention of infective diseases of viral origin in particular; 2) control of chronic noninfectious diseases; 3) prevention of hereditary and congenital diseases. At present, the "classical" recipes for healthy lifestyle are not sufficiently effective under the Russian conditions. There is a need for a fresh comprehensive approach to this problem on the assumption that there is a common pathogenetic link (the effects of active oxygen forms and other free radical products) in the mechanism of the factors of the environment and vital functions. In this connection, the most important task of medicine in the prevention of chronic noninfectious diseases is to enhance the body's resistance to their damaging action. Not only should healthy lifestyle, but also the most important biological reserve, such as cross-adaptation, be placed at the service of prophylaxis. It enhances the body's resistance to different environmental factors during adaptation to one of them. Intermittent hypoxia is the most active adaptive effect. For realization of its preventive effect, it is expedient to set up centers of adaptation medicine and prophylaxis. For prevention of hereditary and congenital diseases, both governmental and medical policies should be to implement a "desired child" programme. PMID- 11055203 TI - American Society of Tropical Medicine and Hygiene 49th annual meeting. October 29 November 2, 2000. Houston, Texas, USA. Abstracts. PMID- 11055204 TI - Cancer of the oral cavity. PMID- 11055205 TI - Annual meeting of the American Epilepsy Society. Los Angeles, California, USA. December 1-6, 2000. Abstracts. PMID- 11055206 TI - American Society for Therapeutic Radiology and Oncology 42nd annual meeting. October 22-26, 2000. Boston, Massachusetts, USA. Abstracts. PMID- 11055207 TI - Pathological Society of Great Britain and Ireland 181st meeting. 12-14 July 2000. Nottingham, United Kingdom. Abstracts. PMID- 11055208 TI - Molecular aspects of arsenic stress. AB - Arsenic produces a variety of stress responses in mammalian cells, including metabolic abnormalities accompanied by growth inhibition and eventually apoptosis. Morphological alterations in cells exposed to arsenic often suggest underlying disruption of cytoskeletal structural elements responsible for cellular integrity, shape, and locomotion. However, specifics of the ultrastructural changes produced by arsenic remain poorly understood. Various tissues and organs differ in their sensitivity to arsenic, with the liver and skin being the most studied. Characteristic skin pathology related to arsenic exposure ranges from hyperkeratotic lesions to squamous-cell carcinomas. However, molecular events in the arsenic-exposed skin still remain to be elucidated. Although mutagenicity of arsenic has not been unequivocally established, recent evidence supports the view that oncogenic mutations do occur, and that only selected enzymes related to DNA replication and repair are affected by arsenic. Sensitivity of the mitotic spindle to arsenic, particularly its organic compounds, underlies the well-documented chromosomal aberrations in arsenic exposed populations. Arsenite-induced stress at the molecular level shares many features with the heat shock response. This includes the differential sensitivity of the stress signal pathway elements to the magnitude of the stress, stressor specific activation of the response elements, and the protective role of the heat shock response. Oxidative stress, the central component of heat shock response, is typical of arsenic-related effects that are, in fact, regarded as the chemical paradigm of heat stress. Similar to heat stress, arsenite induces heat shock proteins (HSPs) of various sizes. The signal cascade triggered by arsenite-like heat stress induces the activity of the mitogen-activated protein (MAP) kinases, extracellular regulated kinase (ERK), c-jun terminal kinase (JNK), and p38. Through the JNK and p38 pathways, arsenite activates the immediate early genes c fos, c-jun, and egr-1, usually activated by various growth factors, cytokines, differentiation signals, and DNA-damaging agents. Like other oxygen radical producing stressors, arsenic induces nitric oxide production at the level of transcriptional activation along with induction of poly(ADP)-ribosylation, NAD depletion, DNA strand breaks, and formation of micronuclei. This review presents an overview of current research on molecular aspects of arsenic stress with an emphasis on the tissue-specific events in humans. It deals with current progress on the understanding of the signal transduction pathways and mechanisms underlying the sensitivity of various species, organs, and tissues to arsenic. PMID- 11055209 TI - Health risks caused by freshwater cyanobacteria in recreational waters. AB - Toxic cyanobacteria are increasingly being perceived as a potential health hazard, particularly in waters used for recreation. A few countries are developing regulations to protect human health from these toxins, and the World Health Organization (WHO) has published both a guideline value for one cyanotoxin in drinking water and a procedural guideline for recreational waters. This article presents an overview of the currently known cyanotoxins and of documented cases of human illnesses attributed to them. It further discusses exposure pathways and approaches to risk management. In this context, the WHO guideline for recreational waters is presented, and monitoring approaches are outlined. PMID- 11055210 TI - [The diagnostic and surgical characteristics of gunshot nonmagnetic intraocular fragments]. AB - Results of treatment of 42 patients with a magnetic intraocular fragments (balls, pistons, capsules) are analyzed and indications for various surgical interventions for removal of fragments with the minimal injury for the eye are defined. The role of echo diagnosis (together with x-ray diagnosis and computer aided tomography) in the selection of operative access to a foreign body and, specifically, echolocation on the operative table during surgery is shown. Combination of informative diagnostic methods with advanced surgical technologies helps save the organ and attain functional results in this extremely grave posttraumatic pathology. PMID- 11055211 TI - [Total posterior cryopexy of the cornea in the treatment of chronic bullous keratopathy]. AB - The proposed method, in contrast to methods used in the treatment of bullous chronic keratopathy with the painful syndrome, suggests cryo applications on the entire posterior surface of the cornea through a short (2.5-3 mm) corneal or scleral incision. Posterior cryopexy can be combined (if necessary) with other reconstructive interventions (anterior vitrectomy, separation of synechiae, reposition of intraocular lens, antiglaucoma surgery). The intervention leads to the formation of a semipermeable fibrocellular film representing a cicatricial biological barrier at the level of endothelial Descemet's membrane for chamber humor. Total posterior cryopexy of the cornea relieves pain due to antiedematous effect, which was noted clinically, by evaluation of corneal permeability by keratopenetrofluorometry, and from improvement of visual acuity. PMID- 11055212 TI - [The use of latex expanders for the formation of a conjunctival space in anophthalmos]. AB - Methods for conjunctival cavity plasty in anophthalmia and their advantages and drawbacks are discussed. The known methods do not always bring about satisfactory results (ensure a stable position of the prosthesis in the cavity). A method for conjunctival cavity formation with latex expanders is proposed. Original designs of expanders for the conjunctival cavity were used. The method is interesting and promising for conjunctival cavity plasty in anophthalmia when the cavity is shrunk. PMID- 11055213 TI - [The characteristics of performing specialized laser keratomileusis after photorefractive keratectomy performed earlier in the correction of myopia]. AB - Ninety-four repeated refraction operations were performed by laser specialized keratomileusis (LASIK) after a previous photorefraction keratectomy in order to correct the complications (undercorrection and regression of the refraction result). LASIK operation can be performed no earlier than 12 months after the first operations if subepithelial corneal fleur develops; the fleur severity is to be about 0.5 points and the optimal thickness of corneal section is 180 mu. Causes of complications of reoperations are discussed. The majority of complications are due to corneal epithelium fixation after photorefraction keratectomy. Refraction and visual results of surgery are presented to characterize its efficiency. PMID- 11055214 TI - [The characteristics of the clinical course of the wound process in the eye]. AB - Clinical course of wound process in the eye was studied in 279 patients with penetrating wounds of different severity. Modern microsurgical and drug treatment of penetrating wounds results in a favorable course in 2/3 (77.4%) of patients, while in the rest 22.6% the course of healing was complicated by intraocular infection (10.7%) and chronic noninfectious posttraumatic uveitis (11.7%). Risk factors of a complicated course were defined, among which of special importance are specific features of the injury and secondary immunodeficiency, clinically manifesting by chronic inflammatory diseases, predominantly of the ENT organs and bronchopulmonary system. PMID- 11055215 TI - [The pathomorphological characteristics of the cataractous lens in glaucoma patients]. AB - Lenses from patients with glaucoma and senile cataract (control) were examined under electron microscope. Ultrastructural changes in the lenses of glaucoma patients differ from changes in senile cataract by a more pronounced thinning of the capsule and development of microdefects in it and in many cases by accumulation of amorphous material on the surface of the posterior capsule adjacent to the vitreous body. This material may be pseudo-exfoliate or a manifestation of stratification of the posterior capsule proper. Development of a pseudomembrane on the surface of posterior capsule is a manifestation of proliferative processes in glaucoma. These features in a certain measure reflect the mechanism of cataract development in glaucoma, which helps plan the measures for preventing it. The status of posterior lenticular capsule in glaucoma is to be taken into consideration when implanting intraocular lenses. PMID- 11055216 TI - [The new interferon inducer Aktipol in the treatment of herpetic keratitis]. AB - Previous experiments demonstrated the efficiency of 0.007% para-aminobenzoic acid (PABA) in the treatment of dendriform herpetic keratitis in rabbits and showed the interferonogenic activity of local administration of PABA in this concentration in ocular tissues. A new drug Aktipol based on 0.007% PABA was developed for the treatment of viral diseases of the eyes. The drug was used in the treatment of patients with surface herpetic keratitis. It was instilled into the conjunctival sac (2 drops 8-10 times a day) or instillations were combined with subconjunctival, parabulbar injections 1-3 times a week, depending on the disease severity and course. Clinical studies demonstrated that Aktipol is effective in the treatment of surface forms of herpetic keratitis (118 patients). Cure was attained in 89.8% patients in 11.3 +/- 0.6 days. Aktipol is superior to IDU (cure in 70.6% patients) and is comparable to that of acyclovir (87.7%). Aktipol accelerates ulcer healing, resorption of infiltration, and cure by 4-5 days in comparison with IDU and is comparable to acyclovir. Aktipol is better tolerated than antiviral IDU agents and acyclovir ointment. PMID- 11055217 TI - [The efficacy of the antioxidant preparation Histochrome in the treatment of hemophthalmos in hypertension and diabetes mellitus]. AB - Hemophthalmos in patients with essential hypertension and diabetes mellitus was treated by antioxidant agent Histochrome manufactured by the Pacific Institute of Bioorganic Chemistry, Far Eastern Division of Academy of Sciences, Vladivostok. The results were analyzed with reference to localization, volume, and period elapsed since hemorrhage into the vitreous body and by hemophthalmos index determined by biomicroscopy and echography of the vitreous body. Histochrome was the most effective in fresh hemophthalmos with 1-2 point index. Visual functions improved in patients with nonproliferative diabetic retinopathy without hemorrhagic syndrome. PMID- 11055218 TI - [New prospects in the treatment of intraocular infection]. AB - Clinical trials were carried out on 55 patients (55 eyes) divided into 2 groups. The main group consisted of 27 patients treated by a therapeutic complex including vitrectomy with ozonized normal saline and the controls (28 pts) were treated by vitrectomy with intravitreal administration of 0.4 mg gentamicin. The diagnosis was made by clinical and laboratory methods. The volume of liquid used in vitrectomy was recorded. Ozonized saline was to be used within the period during which the therapeutic concentration of ozone was retained. Ozonized saline in therapeutic doses is not toxic and in contrast to antibiotics, is characterized by a wide spectrum of effects. Hence, vitrectomy with ozonized normal saline is a new effective method for treating endophthalmitis. PMID- 11055219 TI - [L-arginine in the lacrimal fluid of patients with diabetic retinopathy and the possible role of nitric oxide in the pathogenesis of retinal ischemia]. AB - The concentrations of 23 free amino acids and some other components (ammonium, urea, ethanolamine) were measured by liquid chromatography in the lacrimal fluid of 40 patients with diabetic retinopathy (DR) (15 with the preproliferative stage and 25 with proliferative stage) and 15 normal subjects. The mean level of L arginine in DR patients was 3.5-5 times decreased in comparison with the control (1.2 +/- 0.4 vs. 6.1 +/- 1.5%, p < 0.005). The level of ornithine inversely correlated with the level of L-arginine. The mean level of ornithine in DR patients was increased twofold (12.3 +/- 1.0 vs 6.8 +/- 1.0% in the control, p < 0.005). The mean concentrations of other amino acids virtually did not change. Decreased level of L-arginine in the lacrimal fluid of DR patients may be indicative of increased utilization of this amino acid in retinal ischemia, which, according to the modern concept of DR pathogenesis, can be caused by activation of nitric oxide production. Measurement of L-arginine in the lacrimal fluid can serve as a simple noninvasive and reliable method for early diagnosis of ischemic changes in the retina. PMID- 11055220 TI - [Changes in the objective accommodation indices in myopia and the assessment of the results of accommodation training]. AB - Objective parameters of accommodation (amplitude, time of response, and stability) were evaluated in patients with myopia aged 7-22 years on an AA-2000 Nidek accommodographer. A decrease in accommodation response depended on age and degree of myopia. Rapid accommodation parameters were in high correlation with the relative accommodation reserve. Three groups of patients with different efficiency of accommodation training were distinguished: with positive effect (77.1%), no effect, and paradoxical effect. PMID- 11055221 TI - [The validation of and approaches to the use of immunocorrective agents in contusive eye trauma]. AB - Patients with contusions of the eye of different severity were examined. The concentrations of cytokines (interleukin 1 beta, alpha-interferon, tumor necrosis factor-alpha), levels of immunoglobulins and circulating immune complexes were measured and organ-specific immunity was studied. Severe and medium-severe contusions of the eyeball were associated with immune shifts at a local and systemic levels. Shifts regarded as adaptation compensatory immune response were detected in medium-severe contusions. Severe contusions involve immunity disorders which are characteristic of developing secondary immunodeficiency. Postcontusion hemorrhages are a clinical factor of risk of pathogenetically unfavorable autoimmune reactions. The efficiency of immunocorrective drugs is discussed. PMID- 11055222 TI - [The stimulation of the vitreous body hyalocytes in an experiment]. AB - Loss of vitreous volume as a result of injury or surgical intervention leads to death of the eye. The authors developed a method for hyalocyte culturing on the glass in culture medium. Culture medium was as follows (100 ml): 50 ml medium 199, 30 ml lactate albumin hydrolysate, and 20 ml human serum, group IV. Vitreous cells were collected from rabbit eyes. Primary and secondary growth zones were distinguished in the course of culture growth. Morphologically intact rabbit hyalocytes in the culture were polymorphous. These results may indicate a possibility of transplanted culture development in a live eye. PMID- 11055223 TI - [The role of the osmotic pressure of the blood in the pathogenesis of diabetic changes in the retina]. AB - Based on the regularities of circulation mechanics, the authors hypothesize the role of osmotic pressure in the development of diabetic changes in the retina. A mathematical model of tissue fluid movement based on this hypothesis shows that transmural pressure in choriocapillaries is higher than in exchange vessels of the retina, and therefore unidirectional fluid flow from choriocapillaries to these vessels develops. A sharp increase in glucose concentration is paralleled by an increase in osmotic pressure, which, in turn, creates a negative hydraulic pressure in retinal tissue and leads to overstretching of exchange vessels and appearance of microaneurysms. One of pathogenetically based therapeutic effects of laser coagulation in diabetic retinopathy is creation of "fenestrae" in the retinal pigmented epithelium, facilitating tissue fluid movement. PMID- 11055224 TI - [Efficiency in the dispensary care and surgical treatment of glaucoma patients]. AB - The number of blind and poor-sighted patients with glaucoma remains high, which is due to improper organization of treatment of this patient population, among other things. Surgery is an important treatment modality; its effect is higher in patients with the initial stages of disease, which is seen from our observations. Normal intraocular pressure and high visual acuity are retained for 10 years and longer in patients operated on with stage I glaucoma. In patients with developed and far-advanced glaucoma administered complex treatment after surgery, including retrobulbar injections of trental (0.5 ml, 10 injections per course), cerebrolysin (0.5 ml, 10 injections per course), and electric stimulation of the visual analyzer, the glaucomatous process was arrested, visual acuity improved, and visual field extended. In patients with lens opacity, cataracts were extracted and intraocular lenses were implanted; this improved visual acuity without correction. Due to early diagnosis of glaucoma and properly organized regular check-ups of patients (including complex and timely surgery) vision improved and glaucomatous process stabilized in the majority of patients. PMID- 11055225 TI - [A clinico-pathogenetic classification of tuberculosis of the eye]. AB - Clinical pathogenetic classification is based on the results of many-year observation of a large group of patients in whom the tuberculous etiology of ocular disease was confirmed by a universal approach based on reliable informative diagnostic criteria. Three pathogenetic and 4 clinical forms of ocular tuberculosis were distinguished. The classification for the first time includes ocular involvement in tuberculosis of the central nervous system and tuberculosis of defense system of the eye. Tuberculous diseases of the cornea, sclera, and retinal vessels are presented in combination with uveitis of this or that localization, because they are secondary towards uveitis. The classification includes 4 sections. The diagnosis based on this classification reflects the clinical features of disease and the time course of the process. The classification is diagnostically and therapeutically oriented. It suggests a rational choice of etiological diagnosis and therapy, which is determined by the pathogenetic and clinical forms of ocular tuberculosis. PMID- 11055226 TI - [A case of an ocular form of dirofilariasis]. AB - A rare ophthalmic involvement is described. Patient S. aged 60 years was treated for 6 months without effect for allergic conjunctivitis of the right eye, after which a free moving helminth was detected under the bulbar conjunctiva. The parasite was removed, and symptoms of toxic allergic conjunctivitis rapidly regressed. The helminth was identified as Dirofilaria repens, its size was 120 x 2 mm. PMID- 11055227 TI - [The use of different types of explants in the surgery of retinal detachment]. PMID- 11055228 TI - [The All-Russian Scientific and Practical Conference on Glaucoma: the results and outlook on the eve of the millennium, Moscow, 22-24 November 1999]. PMID- 11055229 TI - 2000 Meeting of the Austrian Society for Hematology and Oncology. Vienna, 4-6 May 2000. Abstracts. PMID- 11055231 TI - British Society for Clinical Cytology 39th annual meeting. 10-12 September 2000. Leicester, United Kingdom, Abstracts. PMID- 11055230 TI - [42nd National Congress of Anesthesia and Resuscitation. Paris, France, 21-24 September 2000. Abstracts]. PMID- 11055232 TI - 10th National Conference of the Inflammation Research Association. 24-28 September 2000. Hot Springs, Virginia, USA. Abstracts. PMID- 11055233 TI - 23rd Congress of Endocrinology and Metabolic Diseases of Turkey joint meeting with the European Federation of Endocrine Societies. Ankara, Turkey, September 7 9, 2000. Abstracts. PMID- 11055234 TI - [German Society of Psychiatry, Psychotherapy and Neurology 2000 Congress: Psychiatry in the year 2000, the European perspective. Aachen, 20-23 September 2000. Abstracts]. PMID- 11055235 TI - 14th Annual North American Cystic Fibrosis Conference. Baltimore, Maryland, USA. November 9-12, 2000. Abstracts. PMID- 11055236 TI - BAR2000. Nordic Biological Alcohol Research Meeting. Oslo, Norway. September 7 10, 2000. Abstracts. PMID- 11055237 TI - Undersea and Hyperbaric Medical Society 33rd annual meeting. June 20-22, 2000. Stockholm, Sweden. Abstracts. PMID- 11055238 TI - Intestinal transport and barrier functions: from molecular defect to clinical disease. International symposium. 8-10 September 2000. Tubingen, Germany. Abstracts. PMID- 11055239 TI - Frequency of hepatitis B virus reactivation in cancer patients undergoing cytotoxic chemotherapy: a prospective study of 626 patients with identification of risk factors. AB - Hepatitis B virus (HBV) reactivation is a well-described complication in cancer patients who receive cytotoxic chemotherapy and may result in varying degrees of liver damage. As chemotherapy is used increasingly in cancer patients, HBV reactivation during cytotoxic treatment may become a more common problem. In lymphoma patients, the incidence of chronic HBV infection has been reported to be 26%, of whom 47% developed HBV reactivation during chemotherapy. However, corresponding data for patients with other malignancies undergoing cytotoxic chemotherapy are not known. In this prospective study, hepatitis B surface antigen (HBsAg) was determined in 626 consecutive cancer patients who received cytotoxic chemotherapy over a 12-month period. Seventy-eight patients (12%) were found to be HBsAg positive. Thirty-four (44%) developed raised alanine transaminase during their course of chemotherapy. In these 34 patients, hepatitis was attributed to HBV reactivation in 15 patients (44%), chronic active HBV infection in 1 patient (3%), hepatitis C infection in 1 patient (3%), malignant hepatic infiltration in 2 patients (6%), and the use of hepatotoxic chemotherapeutic agents in 11 patients (32%). The causes of hepatitis were unknown in 4 patients (12%). HBV reactivation was more likely to develop in patients who were male, younger age, HBeAg seropositive, and those with lymphoma. Presence of malignant hepatic infiltration, baseline pre-treatment alanine transaminase, total bilirubin, and HBV DNA levels did not correlate with the development of HBV reactivation. Of the 15 patients who developed HBV reactivation, antiviral therapy with lamivudine was available and used in 9. There was no HBV-related mortality during chemotherapy. It is concluded that in patients with chronic HBV infection under chemotherapy, HBV reactivation occurs in nearly 20% of them and accounts for 44% of hepatitis cases. The risk factors identified include male sex, younger age, HBeAg seropositive, and the diagnosis of lymphoma. In HBV endemic areas, patients with risk factors for HBV reactivation should be identified prior to receiving cytotoxic treatment and monitored closely. The potential benefit of lamivudine requires further confirmation. PMID- 11055240 TI - Possible role of cytotoxic T cells in acute liver injury in hepatitis C virus cDNA transgenic mice mediated by Cre/loxP system. AB - A line of hepatitis C virus (HCV) transgenic mice was established previously that was mediated by Cre/loxP system using HCV cDNA, including core, E1, E2 and NS2 genes. Intravenous infection of a recombinant adenovirus that expresses Cre DNA recombinase (AxCANCre) induced HCV structural protein expression in the liver of transgenic mice. HCV core protein production and transgene recombination in the mouse liver were serially evaluated after AxCANCre infusion. Core proteins were expressed efficiently and transgene was almost completely recombined in the liver of mice after 3 days and then the levels of both core protein production and transgene recombination decreased continuously for 28 days. However, 30.6% of the transgene recombination remained at 28 days and only 2.7% of core production remained at 28 days after infection. Compared with nontransgenic controls, the serum alanine aminotransferase levels in transgenic mice were significantly higher 10, 14, and 21 days after adenovirus infection. Histological scoring also indicated severe pathological changes in the liver of transgenic mice after adenovirus infection. AxCANCre infusion increased CD8+ lymphocyte infiltration into the liver of transgenic mice compared with that of non-transgenic controls. Furthermore, cytotoxic T lymphocytes (CTLs) isolated from transgenic mice during liver injury were specific for the HCV proteins. These results suggest that HCV structural proteins expressed in the liver of transgenic mice enhanced liver injury. HCV-specific CTLs may be to enhance hepatitis. Thus, the present HCV transgenic mouse model provides a useful model of liver injury due to HCV, and the host immune response may play a pivotal role(s) in the pathogenesis of HCV. PMID- 11055241 TI - MxA gene expression in peripheral blood mononuclear cells from patients infected chronically with hepatitis C virus treated with interferon-alpha. AB - Hepatitis C virus (HCV) infection causes acute and often chronic liver disease. The treatment of choice is interferon-alpha (IFN-alpha). The proportion of patients responding to therapy in terms of a sustained virological response, however, is relatively low. One possible reason for the lack of effectiveness might be neutralization of the drug by host's inhibitory factors. Recent kinetic studies suggested that high doses of IFN-alpha-, especially during the initial phase of therapy, might improve the virological response rates. Eighteen patients infected chronically with HCV were treated with IFN-alpha either at a standard dose (3 x 10(6) to 6 x 10(6) IU IFN-alpha three times weekly) for 6 to 12 months or with an intensified therapy (6 x 10(6) IU IFN-alpha daily) for at least one month. As surrogate parameter for the intracellular effect of the drug, MxA gene expression was quantified in RNA preparations from peripheral blood mononuclear cells. Beta-2-microglobulin (beta2M) concentrations were measured in serum. Serum HCV RNA titers were monitored in parallel. When compared to healthy individuals, untreated patients infected chronically with HCV were found to express 2.8-fold higher amounts of MxA specific transcripts. MxA gene expression and serum beta2M concentrations were found to be induced after administration of IFN-alpha, independent of the virological response not only during the initial phase of the intensified therapy but also over several months during standard therapy. It is concluded from these results that both early non-effectiveness of high dose IFN alpha therapy as well as long-term non-effectiveness of standard therapy are not due to IFN-alpha inhibitory or neutralizing elements in serum. PMID- 11055242 TI - Empty virus-like particle-based enzyme-linked immunosorbent assay for antibodies to hepatitis E virus. AB - Hepatitis E, an enterically transmitted non-A, non-B hepatitis, is a serious viral infection that occasionally causes large epidemics in developing countries. In developed countries, the disease only appears sporadically due to the transmission routes, and it is considered to be less important. The hepatitis E virus (HEV) cannot grow in cultured cells and no reliable assay system has ever been developed. In addition, the present diagnostic are not perfect, and actual rates of HEV infection may be underestimated. Highly purified empty virus-like particles (VLPs) of HEV have been produced by the use of a recombinant baculovirus vector in insect cells. Using these VLPs as an antigen, an enzyme linked immunosorbent assay (ELISA) for antibodies to HEV was developed. A panel of 164 sera that were randomized and coded, and sera collected periodically from three patients with hepatitis E were used for the evaluation. The sensitivity of the assay was shown to be equal to or better than that obtained in previous research that used the same serum panel. The ELISA demonstrated that the serum IgM level of the patients was highest at the onset of the clinical illness and then rapidly decreased. In contrast, a high level of circulating IgG antibody titers lasted for more than 4 years. In Japan, a non-endemic country, the prevalence of the IgG class antibody to HEV in healthy individuals was found to range from 1.9% to 14.1%, depending on the geographical area. Only one out of 900 (0.1%) serum samples was IgM-positive. The IgM class antibody to HEV was detected in 10.8% of non-A, non-B, and non-C acute hepatitis patients in northeast China, whereas none of the patients in Korea had the IgM antibody. The ELISA utilizing the VLPs is sensitive and specific in its detection of the IgM and IgG antibodies to HEV. The ELISA is therefore useful for diagnosing HEV infection and for seroepidemiological study of hepatitis E. PMID- 11055243 TI - Humoral and cellular immune responses to the GB virus C/hepatitis G virus envelope 2 protein. AB - Immune responses to two recombinant envelope 2 (E2) proteins, representing genotypes 1 and 2 of the GB virus C, or the hepatitis G virus (GBV-C/HGV), were studied in mice and in 48 individuals with, or without, chronic, or past GBV C/HGV infection. Immunised mice developed E2-specific antibodies (mean titres, 1:1,167 to 1:9,360), recognising linear antigenic regions and proliferative and IL-2, IL-6 and gammaIFN cytokine responses regardless of the viral genotype. Individuals with past GBV-C infection had E2 antibody titres from 1:1,500 to 1:7,500 that did not recognise the E. coli derived E2 protein or linear antigenic regions. Proliferative E2-specific responses were detected in peripheral blood mononuclear cells from 6/22 (27%) persons with, and in none without GBV-C markers (P<0.05). Thus, E2-specific immune responses are mainly crossreactive between different variants of GBV-C/HGV, although proliferative responses appear to be rare. PMID- 11055244 TI - Prevalence and distribution of human herpesvirus 7 in normal brain. AB - Although it has been recognised that human herpesvirus 7 (HHV-7) establishes latent infection in CD4+ T lymphocytes and productive infection in salivary glands, recent data suggest that its in vivo tropism may be more widespread. In this study, the prevalence and distribution of HHV-7 in brain tissues of 30 consecutive post-mortems were examined by nested polymerase chain reaction. For each post-mortem, 10 fresh autopsy tissue samples were collected respectively from the cerebellum, frontal, temporal, parietal, and occipital lobes of both cerebral hemispheres. These patients were aged from 20-95 years (mean = 61.4, SD = 20.2) with a male:female ratio of 2:1. Three patients died of intracranial haemorrhage, the others died of causes unrelated to the central nervous system. Overall, 5% (15/300) of the brain tissue samples were positive for HHV-7 DNA. The positive rates with respect to anatomical positions were similar (0-3/30). When analysed by patient, 36.7% (11/30) were HHV-7 DNA positive. The viral DNA positive and -negative groups did not show a significant difference in gender or age distribution. The majority (81.8%) of viral DNA-positive patients had HHV-7 DNA detected at only one anatomical position; only two patients had viral DNA detected simultaneously at three anatomical sites. These results suggest that HHV 7 persists in brain tissues of a substantial proportion of the adult population, and in most individuals, its distribution is probably confined to one site rather than pervasive. Further studies to elucidate the role of this ubiquitous virus in neuropathology are warranted. PMID- 11055245 TI - Simultaneous detection of 6 human herpesviruses in cerebrospinal fluid and aqueous fluid by a single PCR using stair primers. AB - A Herpes Consensus allows the simultaneous detection of 6 human herpesviruses: herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), human cytomegalovirus (HCMV), varicella-zoster virus (VZV), Epstein-Barr virus (EBV), and human herpes virus 6 (HHV-6). This technique was used first to examine retrospectively 100 DNA extracts from 95 CSF and 5 aqueous fluids, prepared by treatment by saturated NaCl followed by ethanol precipitation (n = 63) or by simple boiling (n = 37) and stored at -80 degrees C, and secondly to test prospectively 38 CSF samples for which two DNA extracts were prepared with commercially available DNA extraction kits. In all cases, the results were compared with those of an "in-house" PCR. Concordant results between both PCR and the Herpes Consensus techniques were obtained in 61 of 63 DNA extracts prepared by treatment by saturated NaCl (97%) and in only 31 of 37 boiled samples (84%). Both commercially available methods of DNA extraction examined appear to be suitable for Herpes Consensus PCR, although they cannot remove completely PCR inhibitors that must be sought in case of negative results. This preliminary study shows that the Herpes Consensus method should be of value for rapid diagnosis of herpesvirus infections on condition that it is performed on purified DNA extracts. PMID- 11055246 TI - Natural killer cells lyse autologous herpes simplex virus infected targets using cytolytic mechanisms distributed clonotypically. AB - Natural killer (NK) cells have the capability of lysing targets that have down regulated the expression of HLA class I molecules. Herpes simplex virus (HSV) infection results in a profound reduction of HLA class I molecules on the surface of infected cells. For this reason, NK cell populations kill efficiently HSV infected cells. The recent availability of a panel of monoclonal antibodies directed to NK receptors for HLA class I (CD158a, CD158b, anti-p70, anti-p140, and CD94) allowed an accurate dissection of the NK cell subpopulations. Using this approach, the relationship between the expression of NK cell receptors and the capability of lysing HSV-infected cell targets was analyzed at the clonal level. NK cell clones were derived from healthy donors, and cytolytic properties were assayed against HSV-infected autologous fibroblasts. NK cell clones, classified according to the expression of natural killer-cell receptors on their surface, displayed a great heterogeneity of cytolytic properties against HSV infected cells. Nevertheless, a more accurate functional analysis demonstrated not only that HSV infection downregulated the expression of HLA-A and HLA-B and did not modify the expression of HLA-C, but also that NK cell clones expressing the "activating" form of the anti HLA-C NK cell receptor were more cytolytic than other clones. This finding suggests that two different and clonally distributed mechanisms of NK cell activation may be employed by NK cells to kill HSV-infected autologous target cells. PMID- 11055247 TI - Alteration of virus entry mode: a neutralisation mechanism for Dengue-2 virus. AB - Polyclonal antibodies derived from dengue virus immune sera and 3H5 monoclonal antibody showed potent neutralisation effect on dengue-2 virus in the plaque reduction neutralisation assay. This study demonstrated that antibodies present in immune human sera and 3H5 monoclonal antibody neutralised dengue-2 virus by altering the virus entry pathway into cells. In the presence of neutralising antibodies, dengue-2 virus was endocytosed by LLC-MK2 cells. The endocytosis process involved ruffling of antibody-coated virions by cellular pseudopodia and invagination of cell membrane. This mode of entry is atypical as compared to direct fusion of dengue-2 virus with cell membrane in the absence of antibody. The virions were internalised in the form of virion-antibody complexes consisting of single or clumps of virions. After 3 minutes of incubation, neutralised virions were detected in cellular vesicles, and signs of intra-endosomal penetration into cytoplasm were not evident even after a prolonged incubation of 10 minutes, suggesting that viral uncoating was compromised. Vesicle-bound virions were no longer detected after 20 minutes of incubation. In addition, no sign of viral replication was detected in cells infected with "neutralised" virions by immunofluorescence assay. This indicated that internalised virions had been degraded leading to abortive infection. In conclusion, antibodies present in 3H5 monoclonal antibody and human immune sera rendered dengue-2 virus non infective by neutralising the viral fusion site and causing alteration of viral entry mode. Antibodies in immune sera but not 3H5 monoclonal antibody also exerted minimal inhibitory effect on virus binding and internalisation. PMID- 11055248 TI - Further evidence of the absence of measles virus genome sequence in full thickness intestinal specimens from patients with Crohn's disease. AB - Specimens of macroscopically inflamed and normal intestine along with mesenteric lymph nodes were obtained at resection from patients with Crohn's disease. The samples were systematically examined by RT-PCR-nested PCR targeting N, M and H gene regions of the measles virus genome. None of the samples examined gave any evidence of the persistence of measles virus in the intestine of Crohn's disease patients. The study supports previous findings produced by this laboratory and others using highly sensitive measles virus specific PCR diagnostic technology. PMID- 11055249 TI - Picornavirus proteins share antigenic determinants with heat shock proteins 60/65. AB - Immunological cross-reactions between enteroviruses and islet cell autoantigens have been suggested to play a role in the etiopathogenesis of insulin dependent diabetes mellitus (IDDM). In the nonobese diabetic mouse, an autoimmune model of IDDM, one of the reactive beta cell autoantigens is the heat shock protein 60 (HSP60). These studies were prompted by sequence homology discovered between the immunogenic region in HSP60 and two regions in enterovirus capsid proteins, one in the VP1 protein and the other in the VP0, the precursor of VP2 and VP4 proteins. Possible immunological cross-reactions between enterovirus proteins and heat shock proteins were studied by EIA and immunoblotting by using purified virus preparations, viral expression proteins VP1 and VP0, and recombinant HSP60/65 proteins, and corresponding polyclonal antisera. The HSP60/65 family of proteins is highly conserved and there is a striking degree of homology between bacterial and human heat shock proteins. Rabbit antibodies to HSP65 of Mycobacterium bovis that reacted with human HSP60 were also found to recognise capsid protein VP1 of coxsackievirus A9, VP1, and/or VP2 of coxsackievirus B4. Both viruses were also recognised by antisera raised against HSP60 of Chlamydia pneumoniae. In addition to the capsid proteins derived from native virions, antisera to both bacterial HSP proteins recognised expression protein VP1 of coxsackievirus A9. The cross-reactivity was also demonstrated the other way around; antisera to purified virus particles reacted with the HSP 60/65 proteins to some extent. These results suggest that apart from the well-documented sequence homology between the 2C protein of coxsackieviruses and the beta-cell autoantigen glutamic acid decarboxylase, there are other motifs in picornavirus proteins homologous to islet cell autoantigens, which might induce cross-reacting immune responses during picornavirus infections. PMID- 11055250 TI - TT virus infection in patients with chronic liver disease of unknown etiology. AB - The role of a novel virus, designated as TT virus (TTV), as a cause of chronic liver disease has not been well defined. We investigated the prevalence of TTV among 69 patients with chronic liver disease of unknown etiology and 50 volunteer blood donors with normal transaminase levels. TTV DNA was amplified by polymerase chain reaction (PCR) by using two different sets of primers: one based on the sequence of the original N22 clone within the open reading frame 1 (set A) and the other derived from the untranslated region (set B). The prevalence of TTV detected by PCR primers set A only, set B only, and in total (by either set A or B) was 11 (31%), 31 (86%), and 31 (86%) of 36 patients with chronic hepatitis; 2 (40%), 4 (80%), and 4 (80%) of 5 with cirrhosis; 11 (39%), 17 (61%), and 22 (79%) of 28 with hepatocellular carcinoma; and 9 (18%), 39 (78%), and 40 (80%) of 50 volunteer blood donors, respectively. Of the interpretable 25 PCR products amplified with primers set A, 9 were classified as genotype 1a, 10 as genotype 1b, 4 as genotype 2, 1 as genotype 3, and 1 as genotype 4. Molecular evolutionary analysis did not suggest any particular strains of TTV that might be associated with chronic liver disease. The nucleotide sequences of the untranslated region on which PCR primers set B were designed were highly conserved, and the interpretable 22 PCR products amplified with primers set B were not clearly divisible into distinct genotypes. Our findings provided no evidence that TTV is a causative agent of chronic liver disease. PMID- 11055251 TI - Assessment of strategic self-regulation in traumatic brain injury: its relationship to injury severity and psychosocial outcome. AB - Standard neuropsychological tests administered in a constrained and artificial laboratory environment are often insensitive to the real-life deficits faced by patients with traumatic brain injury (TBI). The Revised Strategy Application Test (R-SAT) creates an unstructured environment in the laboratory in which environmental cues and internal habits oppose the most efficient strategy, thus mimicking the real-life situations that are problematic for patients with TBI. In this study, R-SAT performance was related both to severity of TBI (i.e., depth of coma) sustained 2-3 years earlier and to quality of life outcome as assessed by the Sickness Impact Profile. This relationship held after accounting for variance attributable to TBI-related slowing and inattention. These findings support the validity of the R-SAT and suggest that behavioral correlates of quality of life outcome in TBI can be assessed in the laboratory with unstructured tasks. PMID- 11055252 TI - Investigation of executive function change following anterior temporal lobectomy: selective normalization of verbal fluency. AB - The nociferous cortex hypothesis predicts that electrophysiological normalization to distal extratemporal brain regions following anterior temporal lobectomy (ATL) will result in improvements in executive functioning. The present study examined the effects of seizure laterality and seizure control on executive function change. The authors administered the Wisconsin Card Sorting Test (WCST), Trails B, and the Controlled Oral Word Association Test to 174 temporal lobe epilepsy patients who underwent ATL. No significant changes were found on the WCST or Trails B tests, regardless of surgery side or seizure-free status. However, verbal fluency significantly improved in seizure-free patients. Findings were consistent with the nociferous cortex hypothesis suggesting selective executive function improvement following ATL. These findings are discussed in terms of recent research demonstrating extrahippocampal metabolic normalization following surgery. PMID- 11055253 TI - Longitudinal outcomes of Haemophilus influenzae meningitis in school-age children. AB - The purpose of this study was to investigate long-term outcomes of Haemophilus influenzae Type b meningitis in a cohort of school-age survivors. Findings from an initial assessment at mean age 10 years revealed neuropsychological, achievement, and behavioral sequelae in the children with neurologic complications during the acute-phase illness (H. Taylor, C. Schatschneider, & D. Rich, 1992). Here, the cohort was reassessed 1 and 2 years after the initial evaluation to investigate age-related influences on disease sequelae. After excluding children with hearing loss, the sample was divided into 2 groups: an affected group of 39 children with acute-phase neurologic complications and an unaffected group of 73 children without these complications. Growth-curve modeling showed poorer outcomes at the final assessment and less rapid improvement at follow-up for the affected group. Later age at assessment and later age at illness were associated with larger group differences in some outcomes. Results suggest that children with diffuse early brain insults are at risk for later-emerging sequelae. PMID- 11055254 TI - Interhemispheric comparisons in a man with complete forebrain commissurotomy. AB - J. Sergent (1991) claimed that split-brained people are highly accurate in judging which is the larger of 2 circles in opposite visual hemifields but are relatively poor at judging whether circles in the 2 hemifields are of the same size. The discrepancy could be due, at least in part, to an artifact. A split brained man, L.B., was markedly worse than normals at judging which was the larger of 2 circles or the longer of 2 horizontal lines in opposite hemifields, and his performance could be largely accounted for without assuming any interhemispheric transfer. L.B. also judged whether a single flashed line extended further into the left or right hemifield and, as in a previous study (M. C. Corballis, 1995), was strongly biased to respond "right longer." This bias was not observed in the judgments about the circles or the separated lines, suggesting that it is not due to a compression of perceived space in the left hemifield. PMID- 11055255 TI - Contribution of the anteromedial temporal lobes to the evaluation of facial emotion. AB - Amygdala damage can result in impairments in evaluating facial expressions largely specific to fear. In contrast, right-hemisphere cortical lesions result in a more global deficit in facial emotion evaluation. This study addressed these 2 contrasting findings by investigating amygdala and adjacent cortical contributions to the evaluation of facial emotion in 12 patients with right and 11 patients with left unilateral anteromedial temporal lobectomy (RTL and LTL, respectively) and 23 normal controls. RTL but not LTL patients revealed impaired intensity ratings that included but were not exclusive to fear, with the most severe deficits confined to expressions related to affective states of withdrawal avoidance. This suggests that affective hemispheric specializations in cortical function may extend to subcortical limbic regions. In addition, the right amygdala and adjacent cortex may be part of a neural circuit representing facial expressions of withdrawal. PMID- 11055256 TI - Cerebellar cortical degeneration disrupts discrimination learning but not delay or trace classical eyeblink conditioning. AB - The authors investigated classical eyeblink conditioning in a relatively rare patient, B.R., with extensive cerebellar cortical atrophy and marked sparing of the dentate nucleus. Patient B.R.'s ability to acquire and extinguish simple associations (delay and trace conditioning tasks) as well as her ability to acquire more complex associations (temporal and simple discrimination tasks) were examined. There are 2 primary findings from this study. First, B.R. showed normal acquisition and extinction in delay and trace conditioning. Second, she demonstrated a complete inability to learn associative discriminations, even in the case of a simple 2-tone discrimination within the context of a delay paradigm. The latter finding was unexpected because of the sparing of her deep cerebellar nuclei. These data suggest that the cerebellar cortex, or pathways traversing cerebellar cortex, play an important role in classical eyeblink discrimination learning. PMID- 11055257 TI - Verbal working memory and solvent exposure: a positron emission tomography study. AB - Neuropsychological studies have documented frontal dysfunction in patients with a history of exposure to organic solvents. The deficits typically observed in these patients appear to be related to working memory (WM). This study used [15O] water positron emission tomography (PET) to examine the pattern of neural activation during verbal working memory in patients with a history of exposure to solvents. Six individuals with solvent exposure were compared with 6 age- and education matched controls. On the 2 WM tasks examined with PET, with equivalent task performance, participants with solvent exposure demonstrated frontal peaks that were atypical for the tasks, whereas the posterior peaks were typical for the tasks. The results support frontal dysfunction and compensatory use within anterior regions of the WM system in patients with solvent exposure. PMID- 11055258 TI - Effects of divided attention on automatic and controlled components of memory after severe closed-head injury. AB - The relation between attention available at encoding and automatic and consciously controlled aspects of memory was investigated using the process dissociation procedure. Twenty-four severely closed-head injured (CHI) participants (> 1 year postinjury) and 24 matched controls studied word lists in full- and divided-attention conditions. Recall cued with word stems was tested. In contrast to consciously controlled memory, the CHI group did not perform more poorly than the controls in estimates of automatic memory. Furthermore, for both groups, the divided-attention manipulation reduced the controlled estimates of memory, whereas automatic influences remained invariant. These results suggest that automatic memory processes may remain partially immune to the deleterious effects of severe CHI or show recovery by 1 year postinjury. They also indicate that automatic memory processes do not require additional attentional resources following severe CHI. PMID- 11055259 TI - Intact implicit memory for newly formed verbal associations in amnesic patients following single study trials. AB - This study examines the ability of amnesic patients to recover newly formed associations implicitly after a single study trial. Fifteen amnesic patients with various etiologies studied pairs by forming a sentence containing both words. At test, all participants saw 40 intact pairs, 40 rearranged pairs, and 40 new words. All pairs appeared side by side both at study and at test. For the implicit lexical-decision task, 40 nonwords were intermixed with the other pairs, and participants indicated whether both items were words. For the explicit speeded-recognition test, participants were asked to indicate whether both words had appeared at study. Despite being severely impaired on the explicit test, amnesic patients performed like healthy controls on the implicit test, with faster and more accurate responses to intact pairs than to recombined pairs. Contrary to existing theories, the results suggest that amnesic patients can form and retain new associations. PMID- 11055260 TI - Hemispheric interactions during a face--word Stroop-analog task. AB - A Stroop-analog task with faces and words was developed to investigate intrahemispheric and interhemispheric Stroop effects (SEs). Lateralized faces and words were used in an attempt to invoke right- and left-hemispheric specialization, respectively. A prototypical male face, female face, and baby face and the corresponding words man, woman, and baby were presented as congruent or incongruent face-word pairs either to the same visual field (i.e., unilateral presentations) or to separate visual fields (i.e., bilateral presentations). Bidirectional SEs were found. Word distractors interfered with the identification of face targets, and, somewhat surprisingly, an even greater SE was obtained when words were the target and faces were ignored. Laterality effects were most pronounced for bilateral trials, whereby the SE was larger for right-hemisphere than for left-hemisphere target presentations, irrespective of type of target. This finding suggests that the left hemisphere is generally better shielded than the right from interhemispheric interference effects. PMID- 11055261 TI - Intraindividual variability in cognitive performance in older adults: comparison of adults with mild dementia, adults with arthritis, and healthy adults. AB - Intraindividual variability in latency and accuracy of cognitive performance across both trials and occasions was examined in 3 groups of older adults: healthy adults, adults with arthritis, and adults diagnosed with mild dementia. Participants completed 2 reaction-time and 2 episodic-memory tasks on 4 occasions. Results indicated that intraindividual variability in latency was greater in individuals diagnosed with mild dementia than in adults who were neurologically intact, regardless of their health status. Individual differences in variability were stable over time and across cognitive domains. Intraindividual variability was also related to level of performance and was uniquely predictive of neurological status, independent of level of performance. Results suggest that intraindividual variability may be a behavioral indicator of compromised neurological mechanisms. PMID- 11055262 TI - Preattentive and attentive visual search in individuals with hemispatial neglect. AB - Preattentive and attentive visual processing was examined in patients with hemispatial neglect, hemispatial neglect with hemianopia, and control participants. In the preattentive search task, targets possessed a unique feature that was not shared by distractors. In the attentive search task, targets lacked a feature that was present in the distractors. Preattentive search was normal in 3 neglect patients with cortical lesions but not in 2 neglect patients with hemianopia. A 4th neglect patient without hemianopia with a subcortical infarct abnormally used serial search mechanisms in the preattentive task. Neglect patients were characteristically impaired in the contralesional field in the attentive search task. This study demonstrates preserved explicit detection of visual features in cases of hemispatial neglect. PMID- 11055263 TI - Human hepatic metabolism of a novel 2-carboxyindole glycine antagonist for stroke: in vitro-in vivo correlations. AB - 1. The hepatic metabolism of 3-[-2(phenylcarbamoyl) ethenyl]-4,6-dichloroindole-2 carboxylic acid (GV150526), a novel glycine antagonist for stroke, was investigated. 2. After a single intravenous administration of 800 mg GV150526 to healthy volunteers, six metabolites were observed. The major metabolites detected in human plasma have been shown by mass spectrometry to be glucoronides and one sulphate conjugate. 3. After incubation of GV150526 for 6 and 24 h with human liver slices, three glucuronide metabolites were observed. After incubation of GV150526 with pooled human liver microsomes, only one metabolite was observed, with the same molecular weight and HPLC retention time as the synthetic standard GV217053 (GV150526 hydroxylated on the para-position of the phenyl ring). 4. GV150526 hydroxylase enzyme kinetics--a step before sulphation--was determined using pooled human microsomes and was shown to be catalysed by cytochrome P4502C9. Glucuronidation kinetics towards GV150526 using microsomal preparations were also determined. Glucuronidation of GV150526 was observed with UGT1A1 cDNA expressed protein, but not with UGT1A6. 5. The above enzyme kinetic data were used to calculate intrinsic clearance after scaling-up and hepatic clearance were calculated. Since GV150526 has a high plasma protein binding capacity, the effect of GV150526 binding to microsomal protein was determined. Thus, enzyme kinetic data were corrected, plotting the free (unbound) concentration of GV150526 versus enzymatic velocities: apparent Vmax did not alter significantly but apparent Km was approximately 10-fold lower. Correlation of these corrected enzyme kinetic data to predict clearance with in vivo clearance of GV150526 was good when both fu(plasma) and fu(microsomes) were included in the clearance calculations. PMID- 11055264 TI - Sulphation of resveratrol, a natural compound present in wine, and its inhibition by natural flavonoids. AB - 1. Resveratrol, a polyphenolic compound present in grape and wine, has beneficial effects against cancer and protective effects on the cardiovascular system. Resveratrol is sulphated, and the hepatic and duodenal sulphation might limit the bioavailability of this compound. The aim of this study was to see whether natural flavonoids present in wine, fruits and vegetables inhibit the sulphation of resveratrol in the human liver and duodenum. 2. In the liver, IC50 for the inhibition of resveratrol sulphation was 12+/-2 pM (quercetin), 1.0+/-0.04 microM (fisetin), 1.4+/-0.1 microM (myricetin), 2.2+/-0.1 microM (kaempferol) and 2.8+/ 0.2 microM (apigenin). Similarly, in the duodenum, IC50 was 15+/-2 pM (quercetin), 1.3+/-0.1 microM (apigenin), 1.3+/-0.5 microM (fisetin), 2.3+/-0.1 microM (kaempferol) and 2.5+/-0.3 microM (myricetin). 3. The type of inhibition of quercetin on resveratrol sulphation was studied in three liver samples and was determined to be non-competitive and mixed in nature. Km (mean+/-SD; microM) was 0.23+/-0.07 (control), 0.40+/-0.08 (5 pM quercetin) and 0.56+/-0.09 (10 pM quercetin). Vmax (mean+/-SD; pmol min(-1) x mg(-1)) was 99+/-11 (control), 73+/ 15 (5 pM quercetin) and 57 +/- 10 (10 pM quercetin). Kj and Kies estimates (mean+/-SD) were 3.7+/-1.8 pM and 12.1+/-1.7 pM respectively (p = 0.010). 4. Chrysin was a substrate for the sulphotransferase(s) and an assay was developed for measuring the chrysin sulphation rate in human liver. The enzyme followed Michaelis-Menten kinetics and Km and Vmax (mean+/-SD) measured in four livers were 0.29+/-0.07 microM and 43.1+/-1.9 pmol x min(-1) x mg(-1) respectively. 5. Catechin was neither an inhibitor of resveratrol sulphation nor a substrate of sulphotransferase. 6. These results are consistent with the view that many, but not all, flavonoids inhibit the hepatic and duodenal sulphation of resveratrol, and such inhibition might improve the bioavailability of this compound. PMID- 11055265 TI - Biotransformation of the xenoestrogen 4-tert-octylphenol in hepatocytes of rainbow trout (Oncorhynchus mykiss). AB - 1. The biotransformation of a 4-tert-alkylphenol in rainbow trout (Oncorhynchus mykiss) liver was studied to determine the possible fate and activity of these xenoestrogens in fish. 2. Primary trout hepatocytes were incubated with 30 microM 4-(1',1',3',3'-tetramethyl-butyl)[U-14C]phenol (4-tert-octylphenol; 4-t-OP) for up to 3 h. Radiolabelled metabolites were detected by radio-HPLC and the structures were determined by GC-MS analysis of the conjugated or aglycone products. 3. During the first 15 min, 4-t-OP was metabolized at 1.06 pmol x min( 1) x 10(-6) cells. The amount of parent compound metabolized was maximum after a 1-h incubation, when 86% of 4-t-OP was transformed to five other products. 4. The major metabolite comprised 61% of the total recovered radioactivity and was identified as 4-t-OP-beta-glucuronide. 5. The remaining metabolites were formed from the hydroxylation of 4-t-OP on either the C2 (omega-3) or C4 (omega) positions of the alkyl chain, or ortho on the aromatic ring to form a catechol. These oxidized products were also metabolized to glucuronide derivatives conjugated on the phenol ring. 6. The results suggest that oestrogenic alkylphenols could be rapidly transformed in fish liver by both phase I and II metabolic pathways to a number of conjugated products which are unlikely to be active at the oestrogen receptor. PMID- 11055266 TI - Metabolism, excretion and distribution of the flame retardant tetrabromobisphenol A in conventional and bile-duct cannulated rats. AB - 1. 14C-TBBP-A (2,2-bis(4-hydroxy-3,5-dibromophenyl)propane) was administered orally to the conventional and bile-duct cannulated male Sprague-Dawley rat (2.0 mg/kg body weight). Urine, bile and faeces were collected daily for 72 h, and selected tissues were removed for distribution studies. 2. Faeces was the major route of elimination of TBBP-A in the conventional rat (91.7% of dose), and urine was a minor elimination route (0.3%). Enterohepatic circulation was suggested by biliary excretion of 71.3% and faecal excretion of 26.7% of the administered radioactivity in the bile-duct cannulated rat. 3. 14C-labelled residues in tissues were 2% in the conventional rat, and < 1% in the bile-duct cannulated rat. The large and small intestines contained the majority of the tissue 14C activity for both groups of rat. Levels of TBBP-A in liver were < 0.1% and in fat were below the level of quantification. 4. Three metabolites were characterized in 0-24 h bile samples. Glucuronic acid and sulphate ester conjugates were characterized by mass spectrometry. More than 95% of the extractable faecal 14C was identified as parent TBBP-A. 5. Negligible amounts of TBBP-A-derived 14C were associated with carrier proteins in the urine and bile. PMID- 11055267 TI - High conservation of both phase I and II drug-metabolizing activities in cryopreserved rat liver slices. AB - 1. Xenobiotic-metabolizing enzymes, including both cytochrome P450 and phase II conjugating systems, have been characterized in rat liver slices cryopreserved in 12 or 18% dimethylsulphoxide (DMSO). 2. Several cytochrome P450 isoforms in rat liver slices metabolized testosterone to a variety of hydroxylated products. The rates of formation of these same products were well maintained during cryopreservation of the slices in both 12 or 18% DMSO. 3. After cryopreservation of rat liver slices in 18% DMSO, the rates of metabolism of ropivacaine to 3 hydroxyropivacaine, 4-hydroxyropivacaine and PPX (all catalysed by different cytochrome P450 isoforms) were approximately 94, 79 and 82% respectively of the corresponding rates observed with fresh slices. 4. The rates of conjugation of 7 hydroxycoumarin and 1-naphthol by rat liver slices were significantly decreased after cryopreservation in 12% DMSO, but they were maintained when the concentration of this cryopreservant was increased to 18% 5. After cryopreservation in 12% DMSO, the mitochondrial reduction of the tetrazolium salt MTT by rat liver slices was significantly lowered. In contrast, slices cryopreserved in 18% DMSO demonstrated no significant decrease in their capacity to reduce MTT. 6. Thus, in agreement with previous studies, it was found that cytochrome P450-dependent activities are retained after cryopreservation of liver slices. Although phase II-conjugating enzyme activities are more sensitive to cryopreservation, it was shown that increasing the concentration of DMSO present during cryopreservation could circumvent the problem. This modification improves the usefulness of cryopreserved rat liver slices as a tool in drug metabolism studies. PMID- 11055268 TI - Identification of cytochrome P4503A as the major subfamily responsible for the metabolism of roquinimex in man. AB - 1. Roquinimex, a novel immunomodulator, is metabolized in liver microsomes from mouse and rat via cytochrome P450s to four hydroxylated and two demethylated metabolites (R1-6). The study investigated which cytochrome P450 enzyme(s) is responsible for the metabolism of roquinimex in man. 2. Enzyme kinetic analysis demonstrated an apparent Km = 1.28-7.00 mM and Vmax = 50-159 pmol x mg(-1) microsomal protein x min(-1) for the primary metabolites in human liver microsomes. The sum of Cl(int) for the primary pathways was 0.167 microl x mg(-1) microsomal protein x min(-1). 3. A correlation between the formation rate of R1-6 and 6beta-hydroxylation of testosterone was obtained within a panel of liver microsomes from 11 individuals (r2 = 0.72-0.97). Furthermore, significant inhibition (>90%) of roquinimex primary metabolism was demonstrated by ketoconazole and troleandomycin, specific inhibitors of CYP3A4 as well as with anti-CYP3A4 antibodies. Moreover, a similar metabolite pattern was produced from roquinimex by incubation with cDNA-expressed CYP3A4 as by human liver microsomes. 4. In conclusion, these data indicate a major role for CYP3A4 in the formation of roquinimex primary metabolites in human liver microsomes. PMID- 11055269 TI - Biotransformation of 1,8-cineole in the brushtail possum (Trichosurus vulpecula). AB - 1. The metabolic fate of 1,8-cineole was investigated in the brushtail possum. Six possums were fed an artificial diet to which 0.5% 1,8-cineole (wet weight) was added for 2 days. Urine and faeces were collected after the second day. A sample of each was extracted into ethyl acetate and analysed for metabolites. Both free and total levels of metabolites were identified by GC-MS and LC-MS and quantified by GC-MS. 2. The pattern of metabolite excretion was very complex in the brushtail possum. Nineteen metabolites were found in total. Metabolites were categorized into four groups according to the oxidation they had undergone: hydroxycineoles (n = 3), cineolic acids (n = 2), dihydroxycineoles (n = 3) and hydroxycineolic acids (n = 11). No hydroxycineolic acid metabolites have been previously reported as metabolites of 1,8-cineole. 3. Fractional recovery of the ingested dose (2.4 +/- 0.5 g; mean +/- SD) was 0.44 +/- 0.14 (mean +/- SD) in 24 h. Sixty percent of excreted metabolites were hydroxycineolic acids, the most extensively oxidized metabolites. Conjugation with glucuronic acid was inversely related to metabolite polarity, being greatest for hydroxycineoles (41-82%) and minimal for hydroxycineolic acids. 4. Traces of most metabolites were also found in the faeces. PMID- 11055270 TI - Hepatotoxicity of acetaminophen and N-acetyl-p-benzoquinone imine and enhancement by fructose. AB - 1. Although oral administration of 400 mg/kg acetaminophen (APAP) or 1.8-3.4 g/kg sucrose had no effect on serum levels of alanine aminotransferase (ALT) and sorbitol dehydrogenase (SDH), their co-administration resulted in 20-fold increases in ALT/SDH activities. APAP alone (1250 mg/kg, p.o.) caused the elevation hepatotoxicity parameters, but the levels were lower than observed with co-administration of APAP (400 mg/kg) and sucrose (2.6 or 3.4 g/kg). 2. Sucrose associated increase in serum ALT/SDH activities was selective with APAP and not detected with carbon tetrachloride (160 mg/kg, i.p.), D-galactosamine (400 mg/kg, i.p.) or alpha-naphthyl isothiocyanate (100 mg/kg, p.o.). 3. To verify the synergistic mechanism of sucrose, a major reactive intermediate of APAP, N-acetyl p-benzoquinone imine (NAPQI), was given via the portal vein to rat pretreated with sucrose. Clear elevation of ALT/SDH activities was detected in the co treated group. These results, together with an allopurinol-inhibition experiment, suggest the involvement of high-dose sucrose at a step(s) occurring after the metabolic activation of APAP. 4. Co-administration of glucose or fructose as well as sucrose elevated APAP-induced hepatotoxicity parameters in rat. Fructose but not glucose elevated APAP- or NAPQI-induced LDH leakage in a primary hepatocyte system. The results suggest the primary role of fructose is on the sucrose enhancement of APAP toxicity in rat. PMID- 11055271 TI - Prevalence of detectable abnormal prion protein in persons incubating vCJD: plausible incubation periods and cautious inference. AB - BACKGROUND: Both small and large variant Creutzfeldt Jakob disease (vCJD) epidemics are consistent with the current observed incidence. Uncertainty in vCJD projections could potentially be reduced by incorporating information on the prevalence of the infectious agent in persons incubating vCJD. The prospect of vCJD prevalence studies has been raised by detection of abnormal prion protein, thought to be the infectious agent, in appendices and tonsils removed from vCJD patients. Although unlinked anonymous testing of stored operative tissues for abnormal prion protein is very appealing, the design and interpretation of such prevalence studies is complicated by the lack of information on how early in the incubation period of vCJD the abnormal prion protein becomes detectable. METHODS: We simulate a range of vCJD epidemics, consistent with the limited available information on the incidence of vCJD, to illustrate some of the potential problems encountered when interpreting the results from prevalence studies of detectable abnormal prion protein. We assume plausible incubation period distributions and dietary exposure patterns. RESULTS: We demonstrate, in the context of our simulated epidemics, that prevalence studies of detectable abnormal prion protein would require the testing of tens of thousands of operative specimens and, even then, that unlinked anonymous testing positives would be unexpected. PMID- 11055272 TI - Age-period-cohort analyses of breast-, ovarian-, endometrial- and cervical-cancer mortality rates for Caucasian women in the USA. AB - BACKGROUND: Age-period-cohort analyses of US breast-cancer mortality rates reveal an unexpected decrease in risk for women born after 1948. Hormones are thought to play an important role in the aetiology of breast cancer and female gynaecologic cancers, and thus the evaluation of birth-cohort trends for female gynaecologic cancers may shed light on the declining breast-cancer risk among 'baby-boomers'. METHODS: Age-period-cohort analyses are applied to US mortality rates for breast cancer, ovarian cancer, endometrial cancer and cervical cancer from 1950 through 1995. RESULTS: Age-period-cohort analyses provide no clues regarding the declining birth-cohort risk for breast cancer in 'baby-boomers'. The birth-cohort curves for ovarian and endometrial cancers are roughly similar, and largely explained by known risk factors. The calendar-period curve for endometrial cancer reveals increased risk between 1960 and 1980, probably due to increased use of oestrogen replacement therapy. Changes in the birth-cohort curve for cervical cancer reflect, for the most part, changes in sexual activity. An unexpected significant increase in the calendar-period curve for ovarian cancer occurred around 1980. CONCLUSION: Most of the major changes in the calendar-period and birth-cohort curves for breast cancer and female gynaecologic cancers can be explained by documented changes in known risk factors and in medical practice. The decreasing breast-cancer birth-cohort risk among 'baby-boomers' and the increasing ovarian-cancer calendar-period curve after 1980 are recent changes that require further investigation. PMID- 11055273 TI - Back-calculation based on HIV and AIDS registers in Denmark, Norway and Sweden 1977-95 among homosexual men: estimation of absolute rates, incidence rates and prevalence of HIV. AB - BACKGROUND: The Scandinavian countries, Denmark, Norway and Sweden, have established both HIV and AIDS registers to monitor the HIV epidemic. Information in such registers can be used to estimate the number of new HIV infections over time, incidence rates and prevalence. Information from the HIV registers made it possible to study what kind of effects such information had in the estimation process, compared with using information about new AIDS cases only. METHODS: A Markov model back-calculation approach was used. One model incorporated data on cases of both HIV and AIDS. Another model incorporated data on cases of AIDS only. Death or emigration prior to the onset of AIDS and effects of treatment were included in both models. RESULTS: Estimates of absolute rates of HIV for men who have sex with men (MSM) showed a distinct development in each country. Significant differences in incidence rates and prevalence of HIV among MSM were found between Scandinavian countries when information on diagnosed HIV was incorporated. Precision was improved when using both HIV and AIDS diagnosed cases compared with using AIDS cases only. The epidemic in Denmark was more extensive than in the two other countries for the whole study period. DISCUSSION: The results were fairly robust against reasonable variation in the model parameters. The more extensive epidemic in Denmark may have been caused by the homosexual culture denying that HIV was a disease more relevant to them than to others, until the HIV test was publicly available in 1985. PMID- 11055274 TI - Historical HIV prevalence in Edinburgh Prison: a database-linkage study. AB - BACKGROUND: The prevalence of HIV in prisons is often higher than in the surrounding community, because prisons contain a high proportion of injecting drug users (IDUs). Reliable estimation of HIV prevalence in UK prisons only began in the 1990s. Edinburgh, Scotland, experienced a major IDU-related HIV epidemic which began in 1983. We sought retrospectively to estimate HIV prevalence in Edinburgh Prison over the period 1983-94. METHODS: Prison records of all 477 male HIV-positive patients (332 IDUs) in the Edinburgh City Hospital Cohort (believed to include three-quarters of HIV-positive Edinburgh IDUs) were abstracted from Edinburgh Prison. Using this information and the seroconversion intervals of the patients, the number of person-years spent inside the prison by these individuals while HIV-positive was estimated for each calendar month. From this, HIV prevalence was inferred. RESULTS: HIV prevalence in the prison rose from January 1983, as prevalence among Edinburgh IDUs increased, reaching a peak of 8% in December 1984. Prevalence during 1985-86 was 5-6% and then gradually declined, as the surviving HIV-infected IDUs spent less time in the prison. DISCUSSION: These figures are probably underestimates, as some HIV-positive prisoners are not in the cohort. However, the degree of underestimation should not be great and trends over time are reliable. Our estimate for August 1991, 4.1%, compares favourably with the estimate 4.5%, from an anonymous unlinked survey conducted in the prison that month. Prevalence estimates from other UK prisons are reviewed and suggestions made for other uses of database linkage in HIV and IDU epidemiology. PMID- 11055275 TI - One model, several results: the paradox of the Hosmer-Lemeshow goodness-of-fit test for the logistic regression model. AB - BACKGROUND: The Hosmer-Lemeshow test, used extensively to assess the fit of the logistic regression model, is performed by several statistical packages. Recent studies have shown some problems in the use of this test when ties are present. These problems were attributed merely to the test implementation. METHODS: We analysed the order of the observations as an alternative explanation of the problem of ties. Using a data-set of 1393 intensive care unit (ICU) patients we performed the Hosmer-Lemeshow test with all possible subjects dispositions. RESULTS: We obtained about one million different P values, ranging from 0.01 to 0.95. DISCUSSION: It is already known that when the Hosmer-Lemeshow goodness-of fit test is performed with a number of covariate patterns lower than the number of subjects, its result may be inaccurate. We showed that the extent of this problem could be relevant under particular conditions. We also suggest a strategy for estimating the extent of the problem and subsequent interpretation. PMID- 11055276 TI - A comparison of primary and proxy respondent reports of habitual physical activity, using kappa statistics and log-linear models. AB - BACKGROUND: Many epidemiological studies rely in part on proxy informants. There is little published information on the reliability of proxy-respondent reports of physical activity. METHODS: Self-reported data on vigorous and moderate physical activity, from a representative sub-sample of participants in a community-based case-control study of coronary heart disease, were compared with information collected from their next-of-kin. RESULTS: Relative to primary respondents, proxy respondents under-reported activity by approximately 10 percentage points, for both leisure and work-time activity. On a simple three point scale (inactivity/moderate activity/physical activity), 70% of primary-proxy pairs were in exact agreement with regard to leisure time activity and 67% of pairs were in exact agreement on work-time activity. The corresponding values for the weighted kappa statistic were 0.66 [95% confidence interval (CI) 0.59-0.72] and 0.62 (0.54 0.72). Log-linear modelling provided evidence for superior agreement on worktime activity when the proxy was not the primary respondent's spouse. DISCUSSION: Overall levels of primary-proxy respondent agreement on physical activity seem somewhat lower than has been reported for smoking and alcohol-drinking frequency. There seems little reason to prefer spouse proxies when endeavouring to elicit information on work-time physical activity. Log-linear modelling provides an efficient means of exploring covariate effects in observer-agreement studies. PMID- 11055277 TI - Integration of complementary disciplines into the oncology clinic. Part IV. The chiropractor's role in pain management for oncology patients. PMID- 11055278 TI - Integration of complementary disciplines into the oncology clinic. Part V. Nutritional counseling. PMID- 11055279 TI - Integration of complementary disciplines into the oncology clinic. Part VI. Implications for spirituality with oncology patients. PMID- 11055280 TI - Integration of complementary disciplines into the oncology clinic. Part VII. Integrating psychological services with medical treatment for patients with breast cancer. PMID- 11055281 TI - Synthetic biodegradable polymers as orthopedic devices. AB - Polymer scientists, working closely with those in the device and medical fields, have made tremendous advances over the past 30 years in the use of synthetic materials in the body. In this article we will focus on properties of biodegradable polymers which make them ideally suited for orthopedic applications where a permanent implant is not desired. The materials with the greatest history of use are the poly(lactides) and poly(glycolides), and these will be covered in specific detail. The chemistry of the polymers, including synthesis and degradation, the tailoring of properties by proper synthetic controls such as copolymer composition, special requirements for processing and handling, and mechanisms of biodegradation will be covered. An overview of biocompatibility and approved devices of particular interest in orthopedics are also covered. PMID- 11055282 TI - Biomaterial developments for bone tissue engineering. AB - The development of bone tissue engineering is directly related to changes in materials technology. While the inclusion of materials requirements is standard in the design process of engineered bone substitutes, it is also critical to incorporate clinical requirements in order to engineer a clinically relevant device. This review presents the clinical need for bone tissue-engineered alternatives to the present materials used in bone grafting techniques, a status report on clinically available bone tissue-engineering devices, and recent advances in biomaterials research. The discussion of ongoing research includes the current state of osseoactive factors and the delivery of these factors using bioceramics and absorbable biopolymers. Suggestions are also presented as to the desirable design features that would make an engineered device clinically effective. PMID- 11055283 TI - Biodegradable poly(D,L-lactic acid)-poly(ethylene glycol)-monomethyl ether diblock copolymers: structures and surface properties relevant to their use as biomaterials. AB - To obtain biodegradable polymers with variable surface properties for tissue culture applications, poly(ethylene glycol) blocks were attached to poly(lactic acid) blocks in a variety of combinations. The resulting poly(D,L-lactic acid) poly(ethylene glycol)-monomethyl ether (Me.PEG-PLA) diblock copolymers were subject to comprehensive investigations concerning their bulk microstructure and surface properties to evaluate their suitability for drug delivery applications as well as for the manufacture of scaffolds in tissue engineering. Results obtained from 1H-NMR, gel permeation chromatography, wide angle X-ray diffraction and modulated differential scanning calorimetry revealed that the polymer bulk microstructure contains poly(ethylene glycol)-monomethyl ether (Me.PEG) domains segregated from poly(D,L-lactic acid) (PLA) domains varying with the composition of the diblock copolymers. Analysis of the surface of polymer films with atomic force microscopy and X-ray photoelectron spectroscopy indicated that there is a variable amount of Me.PEG chains present on the polymer surface, depending on the polymer composition. It could be shown that the presence of Me.PEG chains in the polymer surface had a suppressive effect on the adsorption of two model peptides (salmon calcitonin and human atrial natriuretic peptide). The possibility to modify polymer bulk microstructure as well as surface properties by variation of the copolymer composition is a prerequisite for their efficient use in the fields of drug delivery and tissue engineering. PMID- 11055284 TI - Hydrolytic degradation of tyrosine-derived polycarbonates, a class of new biomaterials. Part I: study of model compounds. AB - Tyrosine-derived polycarbonates have been identified as promising, degradable polymers for use in orthopedic applications. These polymers are non-toxic, biocompatible, and exhibit good bone apposition when in contact with hard tissue. Tyrosine-derived polycarbonates were designed to incorporate two hydrolytically labile bonds in each repeat unit, a carbonate bond that connects the monomer units and an ester bond connecting a pendent chain. The relative hydrolysis rate of the two bonds will determine the type of degradation products and the degradation pathway of the polymers. In order to study the degradation mechanism of these polycarbonates in more detail, a series of small model compounds were designed that mimic the repeat unit of the polymer. Results obtained from the use of these model compounds suggested that the backbone carbonate bond is hydrolyzed at a faster rate than the pendent chain ester bond. Increasing the length of the alkyl pendent chain lowered the hydrolysis rates of both hydrolyzable linkages, possibly by hindering the access of water molecules to those sites. The hydrolysis rates of both linkages were pH dependent with the lowest rate at pH about 5. The results from this study can be used to explain the degradation behavior of the corresponding polycarbonates as well as their degradation mechanisms. This information is essential when evaluating the utility of tyrosine derived polycarbonates as degradable medical implant materials. PMID- 11055285 TI - Hydrolytic degradation of tyrosine-derived polycarbonates, a class of new biomaterials. Part II: 3-yr study of polymeric devices. AB - The kinetics and mechanisms of in vitro degradation of tyrosine-derived polycarbonates, a new class of polymeric biomaterials, were studied extensively at 37 degrees C. These polymers carry an alkyl ester pendent chain that allows the fine-tuning of the polymer's material properties, its biological interactions with cells and tissue, and its degradation behavior. The polymer carrying an ethyl ester pendent chain, poly(DTE carbonate), has been established as a promising orthopedic implant material, exhibiting bone apposition when in contact with hard tissue. Tyrosine-derived polycarbonates are relatively stable and degrade only very slowly in vitro. Therefore, accelerated studies were conducted at 50 and 65 degrees C to observe the behavior of polymers during the later stages of degradation. Varying the pendent chain length affected the rate of water uptake, initial degradation rate, and physical stability of the polymeric devices. During the 3-yr study, the polymer degraded by random chain cleavage of the carbonate bonds, accompanied by a relatively small amount of pendent chain de esterification. No mass loss was observed during this period at 37 degrees C, but mass loss was readily evident during the accelerated studies at 50 and 65 degrees C. Thus, it is reasonable to assume that mass loss will occur also at 37 degrees C, albeit only after extensive backbone carbonate cleavage and pendent chain ester hydrolysis. The dimension and surface area of the devices influenced the initial degradation rate, but did not significantly affect the overall rate of degradation. No evidence of "acid dumping" or the release of acidic residues found during the degradation of poly(D,L-lactic acid) were observed for this family of tyrosine-derived polycarbonates. PMID- 11055286 TI - Injectable biodegradable polymer composites based on poly(propylene fumarate) crosslinked with poly(ethylene glycol)-dimethacrylate. AB - New injectable, in situ crosslinkable biodegradable polymer composites were investigated consisting of poly(propylene fumarate) (PPF), poly(ethylene glycol) dimethacrylate (PEG-DMA), and beta-tricalcium phosphate (beta-TCP). We examined the effects of the PEG-DMA/PPF double-bond ratio and beta-TCP content on the crosslinking characteristics of the composites including the maximum crosslinking temperature and the gel point, as well as the properties of the crosslinked composites such as the compressive strength and modulus, and the water-holding capacity. The maximum crosslinking temperature was constant averaging 39.7 degrees C for the composite formulations tested. The gel points varied from 8.0 +/- 1.0 to 12.6 +/- 2.5 min and were not affected by the relative amounts of PEG DMA. The compressive strength at yield of PEG-DMA/PPF composites without beta-TCP increased from 5.9 +/- 1.0 to 11.2 +/- 2.2 MPa as the double-bond ratio of PEG DMA/PPF increased from 0.38 to 1.88. An increase in compressive modulus was also observed from 30.2 +/- 3.5 to 58.4 +/- 6.2 MPa for the same range of the PEG DMA/PPF double-bond ratio. Also, the addition of beta-TCP (33 wt%) enhanced the mechanical properties of all composites. The equilibrium water content of networks without beta-TCP increased from 21.7 +/- 0.2 to 30.6 +/- 0.2% for a double-bond ratio of PEG-DMA/PPF ranging from 0.38 to 1.88. However, the mechanical properties of the swollen composites under compression were smaller than the dry ones. These data demonstrate the feasibility of fabricating injectable biodegradable polymer composites with engineered mechanical properties for orthopedic tissue engineering. PMID- 11055287 TI - A review of photocrosslinked polyanhydrides: in situ forming degradable networks. AB - Many orthopaedic injuries could benefit from a high-strength and degradable material with good tissue compatibility. In addition, there is a great clinical need for materials which are easily contoured or placed into complex-shaped defects by a surgeon. We have rationally designed a new class of photocrosslinkable polyanhydride monomers which in situ form high-strength and surface eroding networks of complex geometries. This paper highlights the advantages of these materials for orthopaedic applications and the technique of photopolymerization for reacting these monomers under physiological conditions. The rationale for the material design, photopolymerization kinetics, degradation behavior, and histology in subcutaneous tissue and a model bone defect are presented. PMID- 11055288 TI - Injectable biodegradable materials for orthopedic tissue engineering. AB - The large number of orthopedic procedures performed each year, including many performed arthroscopically, have led to great interest in injectable biodegradable materials for regeneration of bone and cartilage. A variety of materials have been developed for these applications, including ceramics, naturally derived substances and synthetic polymers. These materials demonstrate overall biocompatibility and appropriate mechanical properties, as well as promote tissue formation, thus providing an important step towards minimally invasive orthopedic procedures. This review provides a comparison of these materials based on mechanical properties, biocompatibility and regeneration efficacy. Advantages and disadvantages of each material are explained and design criteria for injectable biodegradable systems are provided. PMID- 11055289 TI - An osteogenic cell culture system to evaluate the cytocompatibility of Osteoset, a calcium sulfate bone void filler. AB - The purpose of the study was to describe a convenient, reliable and quantitative in vitro assay system to assess the cytocompatibility of a calcium sulfate bone filler on two osteogenic cell lines and primary osteoblasts. The hypothesis was that the bone void filler, OsteoSet pellets, would not impact adversely on cell proliferation kinetics or osteogenic potential of selected cells. The hypothesis was tested by standard in vitro methodology of placing OsteoSet pellets either directly in contact with osteogenic cells, or by compartmentalizing within transwell - clear microporous membrane inserts. Data analyses were accomplished with appropriate post hoc statistics (p < or = 0.05). In the presence of the OsteoSet pellets, the cell lines exhibited a decrease in cell proliferation at days 4 and 7, independent of either cell type or tissue culture medium. A decrease in the alkaline phosphatase enzyme activity occurred in the osteogenic cell lines maintained for 9 and 16 days in the presence of the OsteoSet pellets. However, with the exception of the MC3T3E-1 line, no differences were observed with respect to calcium deposition (mineralization) by day 16. Intact human osteocalcin release data for the human-derived OPC1 line and the primary osteoblasts was inconclusive as the OsteoSet pellets may interact with the osteocalcin secreted into the tissue culture medium. The present studies describe a cell culture system to assess the cytocompatibility of bone-graft substitutes with osteogenic cells by compartmentalizing material from direct cell contact (in transwells), and additionally, by evaluating direct cell/biomaterial interactions. PMID- 11055290 TI - Extracellular matrix protein gene expression of bovine chondrocytes cultured on resorbable scaffolds. AB - It has been demonstrated that using cultured chondrocytes that have been seeded onto various biomatrices can enhance the quality of the articular cartilage repair tissue. As tissue-engineering becomes increasingly more complex there is a need to understand how a specific biomaterial may influence gene expression. In this study several commonly used scaffold materials for cartilage tissue engineering were evaluated with respect to their influence on matrix gene expression. Primary cultures of bovine chondrocytes were established in monolayer then seeded onto polylactic acid (PLLA), polyglycolic acid (PGA), collagen matrices. The induction of collagen type I, collagen type II, and aggrecan was observed at various time points on these biomaterials using RT-PCR. The collagen type I gene was upregulated on collagen scaffolds throughout the culture period. PLLA and PGA showed initial induction followed by downregulation. Monolayer culture did not induce collagen I message. Collagen II genes were selectively upregulated after 72 and 96 h post seeding depending the scaffold material. Monolayer culture had strong induction of collagen II. The aggrecan protein was consistently expressed in all scaffold materials cultures and monolayer. PMID- 11055291 TI - Cytotoxicity of poly(96L/4D-lactide): the influence of degradation and sterilization. AB - The cytotoxicity of poly(96L/4D-lactide) (PLA96), and of its accumulated degradation products, was investigated following different sterilization methods and pre-determined heat-accelerated degradation intervals. PLA96 samples sterilized by either steam, ethylene oxide, or gamma irradiation were left untreated (S0 samples), or were degraded for 30 h or 60 h (S30 and S60 samples) at 90 degrees C in water. Extracts of the samples and of the remaining degradation fluids (F30 and F60) were prepared. The toxicity of both unfiltered and filtered extracts was analyzed in a cell growth inhibition (CGI) assay and a lactate dehydrogenase (LDH) leakage assay. Physical analysis of the extracted samples and of the degradation fluids also was performed. The S0 extracts demonstrated no significant CGI. The CGI of the S30 extracts ranged from 37 to 78%, whereas the CGI of the S60 extracts ranged from 6 to 33%. The CGI of the F30 extracts ranged from 19 to 38% and the CGI of the F60 extracts was 98 to 123%. The LDH leakage assay only showed a high response to the unfiltered F60 extracts. Neither sterilization nor filtration appeared to influence the cytotoxicity of the extracts. Particle accumulation, however, might affect cell membrane permeability resulting in LDH leakage. The results of this study suggest that the cytotoxicity of PLA96 is related to the pH and possibly the osmolarity of the tested extracts. The pH and osmolarity, in turn, may depend on variations in the amounts of solubilized lactic acid and oligomers. These variations appear to result from degradation stage-dependent differences in crystallinity, molecular weight and molecular weight distribution of the PLA96 samples. PMID- 11055292 TI - Effects of fluid flow on the in vitro degradation kinetics of biodegradable scaffolds for tissue engineering. AB - Scaffolds fabricated from biodegradable polymers are used extensively in the field of tissue engineering. Many of these scaffolds are subjected to fluid flow, either in vivo or in bioreactors ex vivo. The goal of this study was to examine the effects of fluid flow on the degradation characteristics and kinetics of scaffolds in vitro. Scaffolds with different porosity and permeability values were fabricated using a copolymer of polylactic acid and polyglycolic acid. These scaffolds were subjected to degradation in phosphate buffered saline at 37 degrees C for up to 6 weeks under two test conditions: static and flow (250 microl/min). The porosity of the scaffolds decreased up to 2 weeks and then increased, while the elastic modulus first increased and then decreased over the course of the study. The mass and molecular weight of the scaffolds exhibited a steady decrease up to 6 weeks. The results further indicated that lower the porosity and permeability of the scaffolds, the faster their rate of degradation. Additionally, fluid flow decreased the degradation rate significantly. It is possible that the high rates of degradation observed here were due to autocatalysis of the degradation reaction by the acidic degradation products. PMID- 11055293 TI - Solvent-free fabrication of micro-porous polyurethane amide and polyurethane-urea scaffolds for repair and replacement of the knee-joint meniscus. AB - New porous polyurethane urea and polyurethane amide scaffolds for meniscal reconstruction have been developed in a solvent-free process. As soft segments, copolymers of 50/50 L-lactide/epsilon-caprolactone have been used. After terminating the soft segment with diisocyanates, chain extension was performed with adipic acid and water. Reaction between the isocyanate groups and adipic acid or water provides carbon dioxide and results in a porous polymer. Extra hydroxyl-terminated prepolymer was added in order to regulate the amount of carbon dioxide formed in the foaming reaction. Furthermore, salt crystals ranging in size from 150 to 355 microm were added in order to induce macroporosity. The pore size was regulated by addition of surfactant and by the use of ultrasonic waves. The resulting porous polymer scaffolds exhibit good mechanical properties like a high-compression modulus of 150 kPa. Chain extension with adipic acid results in better mechanical properties due to better defined hard segments. This results from the lower nucleophilicity of carboxylic acids compared to water and alcohols. By adjusting the reaction conditions, materials in which macropores are interconnected by micropores can be obtained. On degradation only non-toxic products will be released; importantly, the materials were obtained by a simple, reproducible and solvent-free procedure. PMID- 11055294 TI - Analysis of retrieved polymer fiber based replacements for the ACL. AB - The present retrospective analysis of 117 surgically excised anterior cruciate ligament (ACL) prostheses was designed to elucidate the etiology and mechanisms of failure of synthetic ligamentous prostheses. They were harvested from young and active patients (26 +/- 7 yrs) at various orthopaedic centers in France between 1983 and 1993. The average duration of implantation of augmentation and replacement prostheses were 21.5 +/- 12.6 and 33.2 +/- 25.3 months, respectively. The principal causes for their excision were ruptures and synovitis. Each ACL prosthesis was examined macroscopically, histologically, and, after tissue removal, by scanning electron microscopy (SEM) to determine the model, manufacturer, surgical technique used at implantation, the extent of healing, the site of rupture, and the morphology of the damaged fibers. Fourteen types of ACL prostheses were analysed, each fabricated using a different combination of polymers, fibers and textile constructions. Consequently, they generated a variety of healing characteristics and mechanical responses in vivo. SEM observations revealed that abrasion of the textile fibers as a result of yarn-on yarn and/or yarn-on-bone contact was a common phenomenon to almost all models, and was the primary cause of prosthetic failure. Healing inside the synthetic ACL was poorly organized, incomplete and unpredictable as the extent of collagenous infiltration into the textile structure did not increase with the duration of implantation. In fact, the collagenous infiltration into certain models appeared to be more detrimental than beneficial since it caused deterioration and fraying of the textile structure rather than serving as a reinforcing matrix around the prosthesis. In conclusion, the present study shows that three mechanisms may be involved in the failure of ACL prostheses: (1) inadequate fiber abrasion resistance against osseous surfaces; (2) flexural and rotational fatigue of the fibers, and (3) loss of integrity of the textile structure due to unpredictable tissue infiltration during healing. PMID- 11055295 TI - The manufacturing techniques of various drug loaded biodegradable poly(lactide-co glycolide) (PLGA) devices. AB - A considerable research has been conducted on drug delivery by biodegradable polymeric devices, following the entry of bioresorbable surgical sutures in the market about two decades ago. Amongst the different classes of biodegradable polymers, the thermoplastic aliphatic poly(esters) like poly(lactide) (PLA), poly(glycolide) (PGA), and especially the copolymer of lactide and glycolide, poly(lactide-co-glycolide) (PLGA) have generated immense interest due to their favorable properties such as good biocompatibility, biodegradability, and mechanical strength. Also, they are easy to formulate into different devices for carrying a variety of drug classes such as vaccines, peptides, proteins, and micromolecules. Also, they have been approved by the Food and Drug Administration (FDA) for drug delivery. This review discusses the various traditional and novel techniques (such as in situ microencapsulation) of preparing various drug loaded PLGA devices, with emphasis on preparing microparticles. Also, certain issues about other related biodegradable polyesters are discussed. PMID- 11055296 TI - Evidence-based librarianship: an overview. AB - OBJECTIVE: To demonstrate how the core characteristics of both evidence-based medicine (EBM) and evidence-based health care (EBHC) can be adapted to health sciences librarianship. METHOD: Narrative review essay involving development of a conceptual framework. The author describes the central features of EBM and EBHC. Following each description of a central feature, the author then suggests ways that this feature applies to health sciences librarianship. RESULTS: First, the decision-making processes of EBM and EBHC are compatible with health sciences librarianship. Second, the EBM and EBHC values of favoring rigorously produced scientific evidence in decision making are congruent with the core values of librarianship. Third, the hierarchical levels of evidence can be applied to librarianship with some modifications. Library researchers currently favor descriptive-survey and case-study methods over systematic reviews, randomized controlled trials, or other higher levels of evidence. The library literature nevertheless contains diverse examples of randomized controlled trials, controlled-comparison studies, and cohort studies conducted by health sciences librarians. CONCLUSIONS: Health sciences librarians are confronted with making many practical decisions. Evidence-based librarianship offers a decision-making framework, which integrates the best available research evidence. By employing this framework and the higher levels of research evidence it promotes, health sciences librarians can lay the foundation for more collaborative and scientific endeavors. PMID- 11055297 TI - Interlibrary loan in primary access libraries: challenging the traditional view. AB - INTRODUCTION: Primary access libraries serve as the foundation of the National Network of Libraries of Medicine (NN/LM) interlibrary loan (ILL) hierarchy, yet few published reports directly address the important role these libraries play in the ILL system. This may reflect the traditional view that small, primary access libraries are largely users of ILL, rather than important contributors to the effectiveness and efficiency of the national ILL system. OBJECTIVE: This study was undertaken to test several commonly held beliefs regarding ILL system use by primary access libraries. HYPOTHESES: Three hypotheses were developed. HI: Colorado and Wyoming primary access libraries comply with the recommended ILL guideline of adhering to a hierarchical structure, emphasizing local borrowing. H2: The closures of two Colorado Council of Medical Librarians (CCML) primary access libraries in 1996 resulted in twenty-three Colorado primary access libraries' borrowing more from their state resource library in 1997. H3: The number of subscriptions held by Colorado and Wyoming primary access libraries is positively correlated with the number of items they loan and negatively correlated with the number of items they borrow. METHODS: The hypotheses were tested using the 1992 and 1997 DOCLINE and OCLC data of fifty-four health sciences libraries, including fifty primary access libraries, two state resource libraries, and two general academic libraries in Colorado and Wyoming. The ILL data were obtained electronically and analyzed using Microsoft Word 98, Microsoft Excel 98, and JMP 3.2.2. RESULTS: CCML primary access libraries comply with the recommended guideline to emphasize local borrowing by supplying each other with the majority of their ILLs, instead of overburdening libraries located at higher levels in the ILL hierarchy (H1). The closures of two CCML primary access libraries appear to have affected the entire ILL system, resulting in a greater volume of ILL activity for the state resource library and other DOCLINE libraries higher up in the ILL hierarchy and highlighting the contribution made by CCML primary access libraries (H2). CCML primary access libraries borrow and lend in amounts that are proportional to their collection size, rather than overtaxing libraries at higher levels in the ILL hierarchy with large numbers of requests (H3). LIMITATIONS: The main limitations of this study were the small sample size and the use of data collected for another purpose, the CCML ILL survey. CONCLUSIONS: The findings suggest that there is little evidence to support several commonly held beliefs regarding ILL system use by primary access libraries. In addition to validating the important contributions made by primary access libraries to the national ILL system, baseline data that can be used to benchmark current practice performance are provided. PMID- 11055298 TI - Public library consumer health information pilot project: results of a National Library of Medicine evaluation. AB - In October 1998, the National Library of Medicine (NLM) launched a pilot project to learn about the role of public libraries in providing health information to the public and to generate information that would assist NLM and the National Network of Libraries of Medicine (NN/LM) in learning how best to work with public libraries in the future. Three regional medical libraries (RMLs), eight resource libraries, and forty-one public libraries or library systems from nine states and the District of Columbia were selected for participation. The pilot project included an evaluation component that was carried out in parallel with project implementation. The evaluation ran through September 1999. The results of the evaluation indicated that participating public librarians were enthusiastic about the training and information materials provided as part of the project and that many public libraries used the materials and conducted their own outreach to local communities and groups. Most libraries applied the modest funds to purchase additional Internet-accessible computers and/or upgrade their health-reference materials. However, few of the participating public libraries had health information centers (although health information was perceived as a top-ten or top-five topic of interest to patrons). Also, the project generated only minimal usage of NLM's consumer health database, known as MEDLINEplus, from the premises of the monitored libraries (patron usage from home or office locations was not tracked). The evaluation results suggested a balanced follow-up by NLM and the NN/LM, with a few carefully selected national activities, complemented by a package of targeted activities that, as of January 2000, are being planned, developed, or implemented. The results also highlighted the importance of building an evaluation component into projects like this one from the outset, to assure that objectives were met and that evaluative information was available on a timely basis, as was the case here. PMID- 11055299 TI - Factors associated with successful answering of clinical questions using an information retrieval system. AB - OBJECTIVES: Despite the growing use of online databases by clinicians, there has been very little research documenting how effectively they are used. This study assessed the ability of medical and nurse-practitioner students to answer clinical questions using an information retrieval system. It also attempted to identify the demographic, experience, cognitive, personality, search mechanics, and user-satisfaction factors associated with successful use of a retrieval system. METHODS: Twenty-nine students completed questionnaires of clinical and computer experience as well as tests of cognitive abilities and personality type. They were then administered three clinical questions to answer in a medical library setting using the MEDLINE database and electronic and print full-text resources. RESULTS: Medical students were able to answer more questions correctly than nurse-practitioner students before and after searching, but both had comparable improvements in the number of correct questions before and after searching. Successful ability to answer questions was also associated with having experience in literature searching and higher standardized test-score percentiles. CONCLUSIONS: Medical and nurse-practitioner students obtained comparable benefits in the ability to answer clinical questions from use of the information retrieval system. Future research must examine strategies that improve successful search and retrieval of clinical questions posed by clinicians in practice. PMID- 11055300 TI - Clarifying the abstracts of systematic literature reviews. AB - BACKGROUND: There is a small body of research on improving the clarity of abstracts in general that is relevant to improving the clarity of abstracts of systematic reviews. OBJECTIVES: To summarize this earlier research and indicate its implications for writing the abstracts of systematic reviews. METHOD: Literature review with commentary on three main features affecting the clarity of abstracts: their language, structure, and typographical presentation. CONCLUSIONS: The abstracts of systematic reviews should be easier to read than the abstracts of medical research articles, as they are targeted at a wider audience. The aims, methods, results, and conclusions of systematic reviews need to be presented in a consistent way to help search and retrieval. The typographic detailing of the abstracts (type-sizes, spacing, and weights) should be planned to help, rather than confuse, the reader. PMID- 11055301 TI - The evolution of rural outreach from Package Library to Grateful Med: introduction to the symposium. AB - Outreach is now a prevailing activity in health sciences libraries. As an introduction to a series of papers on current library outreach to rural communities, this paper traces the evolution of such activities by proponents in health sciences libraries from 1924 to 1992. Definitions of rural and outreach are followed by a consideration of the expanding audience groups. The evolution in approaches covers the package library and enhancements in extension service, library development, circuit librarianship, and self-service arrangements made possible by such programs as the Georgia Interactive Network (GaIN) and Grateful Med. PMID- 11055302 TI - Information needs of rural health professionals: a review of the literature. AB - This review analyzes the existing research on the information needs of rural health professionals and relates it to the broader information-needs literature to establish whether the information needs of rural health professionals differ from those of other health professionals. The analysis of these studies indicates that rural health practitioners appear to have the same basic needs for patient care information as their urban counterparts, and that both groups rely on colleagues and personal libraries as their main sources of information. Rural practitioners, however, tend to make less use of journals and online databases and ask fewer clinical questions; a difference that correlates with geographic and demographic factors. Rural practitioners experience pronounced barriers to information access including lack of time, isolation, inadequate library access, lack of equipment, lack of skills, costs, and inadequate Internet infrastructure. Outreach efforts to this group of underserved health professionals must be sustained to achieve equity in information access and to change information seeking behaviors. PMID- 11055303 TI - Health information outreach: the land-grant mission. AB - Service to the state is one of the core principles of the land-grant mission. This concept of service is also fundamental to a significant number of outreach activities in academic health sciences libraries, particularly those libraries affiliated with the public land-grant universities. The Dana Medical Library at the University of Vermont has a lengthy tradition of outreach to health care providers and health care consumers of the State of Vermont. Building on the foundation of the land-grant institution-which grew out of federal legislation introduced in the mid nineteenth century by Justin Morrill, Vermont's congressional representative--the Dana Medical Library has based its outreach activities on its dedication of service to the state in the promotion of healthy citizens through information dissemination in support of health care delivery. Reengineering library services designed to meet the specific information needs of its diverse clientele, partnering with disparate health care organizations, and relying on fees for service to expand its outreach activities, the Dana Medical Library has redefined the concept of health information outreach for the new millennium. PMID- 11055304 TI - AHEC library services: from circuit rider to virtual librarian. Area Health Education Centers. AB - The North Carolina Area Health Education Centers Library and Information Services (NC AHEC LIS) Network provides library outreach services to rural health care providers in all nine AHEC regions of North Carolina. Over the last twenty-five years, the AHEC and university-based librarians have collaborated to create a model program for support of community-based clinical education and information access for rural health care providers. Through several collaborative projects, they have supported Internet access for rural health clinics. The NC AHEC Digital Library--under development by NC AHEC, University of North Carolina at Chapel Hill, Duke University, East Carolina University, and Wake Forest University--will further extend access to electronic biomedical information and resources to health professionals in a statewide digital library. PMID- 11055305 TI - Library outreach: addressing Utah's "Digital Divide". AB - A "Digital Divide" in information and technological literacy exists in Utah between small hospitals and clinics in rural areas and the larger health care institutions in the major urban area of the state. The goals of the outreach program of the Spencer S. Eccles Health Sciences Library at the University of Utah address solutions to this disparity in partnership with the National Network of Libraries of Medicine-- Midcontinental Region, the Utah Department of Health, and the Utah Area Health Education Centers. In a circuit-rider approach, an outreach librarian offers classes and demonstrations throughout the state that teach information-access skills to health professionals. Provision of traditional library services to unaffiliated health professionals is integrated into the library's daily workload as a component of the outreach program. The paper describes the history, methodology, administration, funding, impact, and results of the program. PMID- 11055306 TI - Health care, information needs, and outreach: reaching Ohio's rural citizens. AB - As a rural state, Ohio has a vital interest in addressing rural health and information needs. NetWellness is a Web-based consumer health information service that focuses on the needs of the residents of Ohio. Health sciences faculty from the state's three Carnegie Research I universities--University of Cincinnati, Case Western Reserve University, and The Ohio State University--create and evaluate content and provide Ask an Expert service to all visitors. Through partnerships at the state and local levels, involving public, private, commercial, and noncommercial organizations, NetWellness has grown from a regional demonstration project in 1995 to a key statewide service. Collaboration with public libraries, complemented by alliances with kindergarten through twelfth grade agencies, makes NetWellness Ohio's essential health information resource. PMID- 11055307 TI - Providing consumer health information in the rural setting: Planetree Health Resource Center's approach. AB - Both lifestyle and geography make the delivery of consumer health information in the rural setting unique. The Planetree Health Resource Center in The Dalles, Oregon, has served the public in a rural setting for the past eight years. It is a community-based consumer health library, affiliated with a small rural hospital, Mid-Columbia Medical Center. One task of providing consumer health information in rural environments is to be in relationship with individuals in the community. Integration into community life is very important for credibility and sustainability. The resource center takes a proactive approach and employs several different outreach efforts to deepen its relationship with community members. It also works hard to foster partnerships for improved health information delivery with other community organizations, including area schools. This paper describes Planetree Health Resource Center's approach to rural outreach. PMID- 11055308 TI - Informationists and librarians. PMID- 11055309 TI - Clinical librarianship. PMID- 11055310 TI - Survey of the pre-school child health surveillance programme in Sweden. AB - A survey of the programme for developmental surveillance in the Child Health Centres (CHCs) in Sweden was performed using a questionnaire administered to the Chief Medical Officers (CMO) of the Child Health Services. The questionnaire asked about methods used for auditory examination, developmental surveillance and identification of disturbances in mother-child interaction. Activities for health promotion concerning breastfeeding, non-smoking and allergy prevention were also queried. Thirty-four CMOs representing 1731 CHCs and 645,000 children answered the questionnaire. The reply rate was 81%. Various methods of auditory examination are offered all infants and children in Sweden. The national guidelines for health supervision are followed fairly closely by all. Screening for disturbances in attention, motor development and perception (DAMP) is performed by all but four districts, with various methods, resources and degrees of co-operation with school health services. Support in mother-child interaction is considered very important and new methods to identify and treat disturbances are gradually introduced. Breastfeeding is encouraged; breast milk is the main source of food for 67% of babies at 4 mo of age. Activities to stop or diminish use of tobacco are ongoing everywhere, as are programmes to identify children at risk of developing allergies and for allergy prevention. Thus, the Child Health Services maintain a high standard and are ambitious about introducing new methods and ideas. PMID- 11055311 TI - Quality of evidence for the present Swedish child health surveillance programme. AB - The present Swedish health surveillance programme includes 15 examinations by a nurse, 5 examinations by a physician, 7 assessments of development, 2 assessments of hearing and 1 assessment of visual acuity. The WHO criteria for evaluation of screening programmes can be applied to the Swedish health surveillance programme. These criteria state that the health problem must be important, that there should be an early phase during which the condition is only detectable by medical professionals and that treatment at an early phase should favourably affect the prognosis. The quality of evidence for fulfilment of these criteria has been graded I-III. Grade II-2 refers to evidence obtained from well-designed cohort or case-control analytical studies. The following disorders might be affected by health surveillance at child health centres: amblyopia, ADHD/DAMP, failure to thrive, cerebral palsy, congenital heart failure, congenital luxation of hip, hearing impairment (severe or moderate), mental retardation, retentio testis and hydrocephalus. None of these conditions fulfils the WHO criteria with quality of evidence grade II-2 or better. Thus the evidence for the present Swedish health surveillance programmed is problematic. PMID- 11055312 TI - How do we identify hearing impairment in early childhood? AB - This review discusses the need for universal neonatal hearing screening. Historical background is given concerning conventional childhood hearing screening programmes in western countries. Direct studies on the effects of very early habilitation programmes on speech and language development are cited. Measurement of otoacoustic emissions (OAE) as a tool for neonatal hearing screening is presented. The state of neonatal hearing screening programmes in the US and in Europe, particularly in Sweden, is discussed. PMID- 11055313 TI - Early identification of children with developmental disabilities. AB - This paper provides an overview of research into early identification of children with developmental disabilities in child healthcare, especially those disabilities related to cognitive impairment. The review covers the following related topics: definition of the target group, the predictive value of developmental screening instruments and psychomotor tests, risk indexes, early intervention and evaluation of developmental screening programmes. Empirical research into child development and the predictive value of developmental tests is extensive. However, proportionally few, mostly cohort or case-control, studies focusing on evaluation of developmental screening programmes conducted within a clinical setting were found. Some sensitivity and most specificity rates reported fell within what is considered acceptable for developmental screening performed in the pre-school years, i.e. a sensitivity of more than 70% and a specificity between 70% and 80%. Overall, between 1-6% of the children screened were identified. Typically, most children with severe disabilities were identified prior to the screening or excluded from the studies reviewed. The shortcomings of developmental screening (instruments) and difficulties in early identification are discussed. PMID- 11055314 TI - The role of the Child Health Services in the identification of children with possible attention deficit hyperactivity disorder/deficits in attention, motor control and perception (ADHD/DAMP). AB - Literature concerning the role of the Child Health Services in the identification of children with possible Attention Deficit Hyperactivity Disorder/Deficits in Attention, Motor Control and Perception (ADHD/DAMP) is summarized in order to establish a background for evaluation, discussion and conclusion. PMID- 11055315 TI - The role of the Child Health Services in promoting mental health: an introduction. AB - This paper gives an introduction to the psychosocial work in the Swedish Child Health Services (CHS). There are substantial problems in defining and evaluating the preventive mental health work of the CHS. The issues raised include: why early preventive intervention is important; the promotion of parental mental health as an aim of the CHS; how the CHS can increase parenting knowledge and skills; what evidence there is about intervention among target groups; and finally, how recent research knowledge can be applied in the CHS. PMID- 11055316 TI - A review of interventions in the parent-child relationship informed by attachment theory. AB - Bowlby's attachment theory has inspired a dramatic shift in the way we understand the development of the early relationship between infant and caregiver(s). Though almost all infants develop an attachment relationship to their primary caregiver(s), not all of them are able to use their caregiver(s) as a secure base or haven of safety from which to explore the world. Mary Ainsworth was the first to describe in detail the aspects of the caregiving system that are most important for the development of the attachment relationship. During the last decade several studies have started to evaluate the possibility that insensitive and perhaps even inadequate parenting can be effectively ameliorated by interventions. The present article sketches the theory behind attachment-based interventions and reviews the evidence for the effectiveness of such interventions. Finally, the article gives examples of successful preventive as well as therapeutic interventions. PMID- 11055317 TI - Secondary prevention in child health: effects of psychological intervention, particularly home visitation, on children's development and other outcome variables. AB - This paper reviews interventions targeting socially deprived families, families with low birthweight/premature children, and some other problems (child abuse, sensitivity/attachment, postnatal depression). Conclusions are mainly based on randomized controlled trials. Earlier reviews in the field have emphasized the importance of intensive, enduring home visitation and of early education programmes for young children. Home visitation may positively effect several outcomes, including health behaviour, child safety and stimulation. Rates of child abuse and neglect have proven difficult to influence, but home visitation may result in other gains such as fewer accidents and serious injuries, and greater home safety. The cognitive development of low birthweight and premature children may be positively influenced by home visitation, particularly in combination with an early stimulation programme in the neonatal unit and pre school placement. Postnatally depressed mothers have been shown to improve substantially from nurse counselling once a week for 6-8 wk. It is suggested that home visitation should be tried on a systematic basis, and that early pre-school experiences should be offered to children in different risk situations. Child Health Centres should introduce a screening programme for postnatal depression. Specialist child health units should be encouraged. PMID- 11055318 TI - Health promotion at Swedish Child Health Centres. AB - Effective health promotion intervention is understood to modify appropriate risk and protective factors. The relevance of such interventions for the Child Health Service (CHS) was examined in three steps. In the first step, the six most important public health problems in the target group that might be affected by CHS interventions were identified. The health problems include sudden infant death syndrome (SIDS), mental health problems, injuries, infections, asthma and allergic and nutritional disorders. In the second step, three groups of modifiable determinants were identified: risk and protective factors for (i) SIDS, asthma and allergic disorders and airway tract infections (determinants: breastfeeding, environmental tobacco smoke, sleeping non-prone and lacking indoor ventilation). (ii) injuries and (iii) mental health problems. In the third step, evidence for the preventive measure's ability to alleviate the effects of these risk factors was scrutinized. Evidence was found for effectiveness of CHS interventions aiming at decrease of environmental tobacco smoke. Evidence was also found for injury prevention provided that the intervention is a part of a wider community effort, and possibly also if it is very focused. Requirements for mental health promotion are also at hand. Thus, effective health promotion via CHS interventions is feasible. PMID- 11055319 TI - The role of the Swedish Child Health Services in breastfeeding promotion. AB - Sweden has one of the highest breastfeeding incidence and duration rates among industrialized countries. Although the Child Health Services offer breastfeeding support to all mothers, there are geographical differences in breastfeeding frequency at different ages. The aims of this study were to describe the present activities in the Child Health Services regarding breastfeeding promotion and to find research evidence regarding interventions. Thirty-three out of 42 healthcare districts replied to a questionnaire. Differences were found in the transfer of responsibility for newborn infants from hospital to Child Health Centres, criteria for and timing of home visits and recommendations regarding introduction of supplementary food and breastfeeding education for parents and professionals. There were also regional differences in breastfeeding statistics and follow-up periods. The following recommendations were made on the basis of the survey and relevant literature: transfer of responsibility for newborn infants must guarantee follow-up of all mother-infant pairs; uniform breastfeeding assessment and documentation must be established; all mother-infant pairs must be offered early home visits, continued on a regular basis by health visitors; drop-in consultations must be established; a telephone hotline must be set up; for preventive purposes, growth charts must be used based on breastfed infants; evidence-based guidelines for the introduction of other foods must be followed; information must be provided in parent groups; breastfeeding statistics must use WHO definitions; polyclinics must be available for service to mothers/infants after early discharge and as resources for Child Health Centres; Child and Maternal Health Centres must collaborate; quality assurance programs must be established; breastfeeding courses must be offered in the under- and postgraduate training of professionals; compulsory in-service education must be offered; lactation consultant training must be offered at the university level; and lactation consultant positions must be established. PMID- 11055320 TI - How to prevent exposure to tobacco smoke among small children: a literature review. AB - There are many reviews of current knowledge about smoking cessation in general within the health service, which also contain guidelines about smoking cessation during pregnancy. Our aim was to review methods in child healthcare for preventing the exposure of children to tobacco smoke. Since passive smoking starts during pregnancy, we also considered methods in antenatal care. We did a search for relevant articles, especially on randomized, controlled trials, in various databases, chiefly Medline. We mainly analysed studies from the last 10 y, concentrating on the actual interventions. In antenatal care the greatest effect comes from interventions based on behavioural strategies. These can lead to a doubling of the number of women who stop smoking during pregnancy. Purely factual information, on the other hand, has no great effect. The studies in child healthcare analysed here show that decisive factors for children not being exposed to passive smoking are a concentration on strengthening the parents' faith in their ability to create a smoke-free environment, and on behavioural strategies to achieve this goal, but not primarily on getting the parents to stop smoking. However, we need further studies of different types of interventions, geared to smokers with small children, before more specific recommendations can be given as to how child healthcare should design its tobacco-preventive work. PMID- 11055321 TI - Is prevention of allergy and asthma possible? AB - Allergy and asthma are common diseases today. Ways to stop the ongoing increase in incidence of these diseases are highly desired. In this review the scientific basis for allergy and asthma prevention is discussed. RESULTS: Although secondary preventive measures are generally regarded as important, the views regarding the possibility of primary prevention vary. There is today only weak evidence that breastfeeding may reduce the risk of developing allergy, and if it does the effect is limited in degree and duration. There is some evidence that prolonged breastfeeding reduces the risk of developing asthma, at least in allergy-risk infants. There is also some evidence indicating that withholding solid foods in children at risk may have a beneficial effect. A dose-dependent risk of becoming sensitized to house dust mite and pet animal allergens has clearly been shown, but to what extent this sensitization is combined with asthma or clinical allergy is less well elucidated. Passive smoking increases the risk of wheezing problems and the indoor environment in damp houses seems to act synergistically with passive smoke. CONCLUSION: Primary prevention is rarely possible, but the risk of developing asthma and allergy may to some extent be reduced by taking certain measures, such as avoidance of tobacco smoke and damp houses. Breastfeeding seems to reduce the risk of wheezing, but as allergy-preventive measures, breastfeeding and avoiding pet animals have only marginal effects. PMID- 11055322 TI - The importance of the Child Health Services to the health of children: summary of the state-of-the-art document from the Sigtuna conference on Child Health Services with a view to the future. AB - In September 1999, a state-of-the-art conference was held in Sigtuna outside Stockholm, Sweden. The subject of the conference was the future of the Child Health Services. The approximately 40 participants included researchers from all professions with a link to child healthcare. The conference was prepared by a working committee consisting of Claes Sundelin (chairman), Uppsala, Sven Bremberg, Huddinge, Gisela Dahlqvist, Umea, Kerstin Hedberg Nyqvist, Uppsala, Anders Hakansson, Malmo, Gunilla Lindmark, Uppsala, Birgitta Wickberg, Goteborg and Maria Nystrom Peck (secretary), Stockholm. The state-of-the-art report has recently been published by the National Council for Medical Research and is briefly summarized below. PMID- 11055323 TI - Safety and pharmacokinetic study with escalating doses of 3-acetyl-7-oxo dehydroepiandrosterone in healthy male volunteers. AB - OBJECTIVES: To evaluate the safety and pharmacokinetics of 3-acetyl-7-oxo-DHEA (3beta-acetoxyandrost-5-ene-7,17-dione) given orally. DESIGN: A randomized, double blind, placebo-controlled, escalating dose study. SETTING: The Chicago Center for Clinical Research. PARTICIPANTS: Twenty-two healthy men. STUDY METHOD: The participants received placebo (n = 6) or 3-acetyl-7-oxo-DHEA (n = 16) at 50 mg/d for 7 days followed by a 7-day washout; 100 mg/d for 7 days followed by a 7 day washout; and 200 mg/d for 28 days. OUTCOME MEASURES: Safety parameters, evaluated at each dose level, included measurement of total testosterone, free testosterone, dihydrotestosterone, estradiol, cortisol, thyroxin and insulin levels. Analyses for 7-oxo-DHEA-3beta-sulfate (DHEA-S), the only detectable metabolic product of the administered steroid, were conducted on plasma drawn from all subjects at 0.25, 0.5, 1, 2, 4, 6 and 12 hours after the final 100 mg dose of 3beta-acetyl-7-oxo-DHEA. RESULTS: There were no differences in the clinical laboratory values or in reported minor adverse experiences, between treatment and placebo groups. In general, blood hormone concentrations were unaffected by the treatment with 3beta-acetyl-7-oxo-DHEA and remained within the normal range. No changes in vital signs, blood chemistry or urinalysis occurred during treatment with 3beta-acetyl-7-oxo-DHEA compared to placebo. The administered steroid was not detected in the blood but was rapidly converted to 7 oxo-DHEA-S, the concentrations of which were proportional to dose. This steroid sulfate did not accumulate; plasma concentrations 12 hours after the 3beta-acetyl 7-oxo-DHEA dose at 7 and 28 days on the 200 mg/d dose were 15.8 and 16.3 microg/L respectively. The mean time to peak plasma level of 7-oxo-DHEA-S was 2.2 hours; the mean half life was 2.17 hours. The apparent clearance averaged 172 L/h, and the apparent mean volume of distribution was 540 L. CONCLUSION: These results indicate that 3beta-acetyl-7-oxo-DHEA is safe and well tolerated in normal healthy men at doses up to 200 mg/d for 4 weeks. PMID- 11055324 TI - Herbal tea in the treatment of diabetes mellitus. AB - OBJECTIVE: To evaluate the effects of a native herbal tea in patients with type 2 diabetes. DESIGN: Randomized, placebo-controlled, single-blind study. SETTING: The Metabolic Centre at the University of Alberta Hospitals. SUBJECTS: Forty volunteers with type 2 diabetes. INTERVENTIONS: After a 1 month "run-in" period, subjects drank 250 mL/d of either the herbal tea or a placebo tea for 10 days, and were followed up for a further 4 weeks. OUTCOME MEASURES: A responder analysis defined as a 10% change in mean blood glucose levels based on 4 capillary glucose readings daily. Secondary end points included changes in HbA1c, fructosamine and response to a meal challenge using Ensure. RESULTS: The responder analysis showed no benefit from the herbal tea. Fructosamine levels before and after tea therapy decreased significantly in both study groups. Mean HbA1c levels and incremental areas under the glucose curve (AUC) in the meal challenge did not change in either study group. These data were reanalysed in hyperglycemic subjects with HbA1c levels greater than 120% of normal. The responder analysis and HbA1c levels did not change in either group. Mean (and standard deviation) fructosamine levels, before and after tea therapy, were significantly lower in the herbal tea group than in the placebo tea group (361 [98] versus 338 [100] micromol/L, p < 0.01 compared with 338 [60] versus 323 [49] micromol/L, p = 0.08). In the hyperglycemic subgroup the mean AUC during the meal challenge, before versus after tea therapy, was 776 (369) versus 639 (331) mmol/L (p = 0.22) in the herbal tea group and 433 (125) versus 420 (173) mmol/L (p = 0.90) in the placebo group. CONCLUSIONS: Although the responder analysis failed to show an effect of the herbal tea, the data suggest there may be a short-term benefit from the tea in subjects with poor glycemic control. PMID- 11055325 TI - A shortened course of anticoagulation to treat deep venous thrombosis after total joint arthroplasty. AB - OBJECTIVE: To study whether the course of anticoagulation therapy in patients who have deep venous thrombosis (DVT) after total joint arthroplasty can be shortened with a minimal risk of recurrence. DESIGN: A case series. SETTING: Kingston General Hospital, a university-affiliated tertiary care centre. PATIENTS: Eleven patients who were found to have DVT after total hip or knee arthroplasty on colour duplex Doppler ultrasonography, who fulfilled the study criteria and gave their informed consent. Exclusion criteria included chronic predisposing factors for thromboembolic disease, revision arthroplasty and a previous DVT. INTERVENTIONS: Anticoagulation with warfarin to achieve an International Normalized Ratio of 2.0 to 2.5, adjusted 3 times a week until resolution of the DVT by duplex ultrasonography. Clinical and ultrasonographic evaluation at 1 year to monitor DVT recurrence. OUTCOME MEASURES: Resolution and recurrence of the DVT. RESULTS: All patients showed resolution of the DVT at their first follow-up ultrasonography (mean 34 days post-operatively). There was no clinical or ultrasonographic evidence of recurrence at 1 year. CONCLUSION: Further study of a shorter course of anticoagulation therapy in patients who suffer DVT after joint arthroplasty should be considered. PMID- 11055326 TI - Bcl-2 intersects the NFkappaB signalling pathway and suppresses apoptosis in ventricular myocytes. AB - As a first step toward identifying putative regulators of apoptosis in the heart, the impact of the anti-apoptosis protein Bcl-2 (B-cell lymphoma gene) on the NFkappaB (nuclear factor kappa beta) signalling pathway in suppressing apoptosis in ventricular myocytes was studied. The data indicate that adenovirus-mediated delivery of Bcl-2 resulted in a significant increase in NFkappaB-dependent DNA binding and NFkappaB-directed gene transcription. No change in NFkappaB protein content was observed in myocytes expressing Bcl-2. Moreover, the Bcl-2-mediated NFkappaB activation was found to be related to changes in the activity of the NFkappaB regulatory protein IkappaBalpha (inhibitor of kappa beta). In this regard, a marked reduction in IkappaBalpha protein content was observed in ventricular myocytes expressing Bcl-2. The mode by which Bcl-2 regulates IkappaBalpha was related to the N-terminal phosphorylation and degradation of IkappaBalpha by the proteasome since an N-terminal deletion mutant of IkappaBalpha or the proteasome inhibitor lactacystin abrogated Bcl-2's inhibitory effects on IkappaBalpha and prevented NFkappaB activation. Furthermore, adenovirus-mediated delivery of a phosphorylation defective form of IkappaBalpha rendered ventricular myocytes incapable of NFkappaB activation and susceptible to tumour necrosis factor alpha-mediated apoptosis. Moreover, Bcl-2's anti-apoptotic function was lost in cells defective for NFkappaB activation. The data provide evidence for a link between Bcl-2 and the NFkappaB signalling pathway for the suppression of apoptosis in ventricular myocytes. PMID- 11055327 TI - World Alzheimer Congress 2000. PMID- 11055328 TI - Solution- and solid-phase synthesis of peptide-substituted thiazolidinediones as potential PPAR ligands. AB - Solution- and solid-phase methods for the preparation of peptide-substituted thiazolidinediones have been developed as an approach towards the preparation of a library of these compounds as potential ligands for the peroxisome proliferator activated receptors (PPARs). PMID- 11055329 TI - Synthesis and antiamoebic activity of new cyclooctadiene ruthenium(II) complexes with 2-acetylpyridine and benzimidazole derivatives. AB - Reaction of [Ru(eta4-C8H12) (CH3CN)2 Cl2] with 2-(2-pyridyl) benzimidazole or Schiff bases derived from 2-acetylpyridine and S-methyldithiocarbazate, S benzyldithiocarbazate and thiosemicarbazide leads to form new complexes of the type [Ru(eta4-C8H12)(L)Cl2] (where L=ligand). In vitro, most of the compounds exhibited potent activity and the Ru derivatives 1a [Ru(eta4-C8H12)(2-Acpy SMDT)Cl2], 2a [Ru(eta4-C8H12)(2-Acpy-SBDT)Cl2] and 3a [Ru(eta4-CsH12)(2-Acpy TSC)Cl2] were found more active than metronidazole against (HK-9) strain of Entamoeba histolytica. PMID- 11055330 TI - Design and synthesis of piperidinyl piperidine analogues as potent and selective M2 muscarinic receptor antagonists. AB - Identification of a number of highly potent M2 receptor antagonists with >100 fold selectivity against the M1 and M3 receptor subtypes is described. In the rat microdialysis assay, this series of compounds showed pronounced enhancement of brain acetylcholine release after oral administration. PMID- 11055331 TI - Differential antibacterial activity of moenomycin analogues on gram-positive bacteria. AB - The moenomycin trisaccharide degradation product and synthetic disaccharide analogues based on the disaccharide core were bactericidal to gram-positive bacteria, inhibited lipid II polymerization, and inhibited cell wall synthesis in Enterococcus faecalis. Truncating moenomycin to the trisaccharide, and building upon the core disaccharide have both led to molecules possessing properties not shared with their respective parent structures. PMID- 11055332 TI - Diphenyl sulfoxides as selective antagonists of the muscarinic M2 receptor. AB - Structure activity studies on [4-(phenylsulfonyl)phenyl]methylpiperazine led to the discovery of 4-cyclohexyl-alpha-[4-[[4-methoxyphenyl(S)-sufinyl]phenyl]-1-pi perazineacetonitrile, 1, an M2 selective muscarinic antagonist. Affinity at the cloned human M2 receptor was 2.7 nM; the M1/M2 selectivity is 40-fold. PMID- 11055333 TI - Selective kappa-opioid antagonists related to naltrindole. Effect of side-chain spacer in the 5'-amidinoalkyl series. AB - The study of kappa-opioid receptor function in vivo has been hampered by the limited choice of selective kappa-antagonists. Recently discovered kappa antagonists have not yet been utilised in vivo. We here report the synthesis and in vitro evaluation of a new amidine derivative of naltrindole. It showed substantially greater kappa-selectivity in binding assays than known analogues with shorter spacer in the amidine side chain. This indicates that in nor-BNI and related selective kappa-antagonists, the second basic centre may not be ideally located. PMID- 11055334 TI - Analogues of SB-203207 as inhibitors of tRNA synthetases. AB - SB-203207 and 10 analogues have been prepared, by elaboration of altemicidin, and evaluated as inhibitors of isoleucyl, leucyl and valyl tRNA synthetases (IRS, LRS, and VRS, respectively). Substituting the isoleucine residue of SB-203207 with leucine and valine increased the potency of inhibition of LRS and VRS, respectively. The leucine derivative showed low level antibacterial activity, while several of the compounds inhibited IRS from Staphylococcus aureus WCUH29 more strongly than rat liver IRS. PMID- 11055335 TI - Highly potent and selective peptide-based inhibitors of the hepatitis C virus serine protease: towards smaller inhibitors. AB - Structure-activity studies on a hexapeptide N-terminal cleavage product of a dodecamer substrate led to the identification of very potent and highly specific inhibitors of the HCV NS3 protease/NS4A cofactor peptide complex. The largest increase in potency was accomplished by the introduction of a (4R)-naphthalen-1 yl-4-methoxy substituent to the P2 proline. N-Terminal truncation resulted in tetrapeptides containing a C-terminal carboxylic acid, which exhibited low micromolar activity against the HCV serine protease. PMID- 11055336 TI - NMR line-broadening and transferred NOESY as a medicinal chemistry tool for studying inhibitors of the hepatitis C virus NS3 protease domain. AB - This work describes the use of NMR as a medicinal chemistry tool for better understanding the binding characteristics of inhibitors of the HCV NS3 protease. The protease-bound structure of a tetrapeptide-like inhibitor that has an acid C terminus, a norvaline at P1 and a naphthylmethoxy proline at P2 is described. Conformational comparisons are made with a similar compound having a 1-amino cyclopropylcarboxylic acid at P1 and with a hexapeptide inhibitor. Differences between the free and bound states are identified. 19F NMR also helped in determining that a single complex is observed when an inhibitor is added to the protease at a 1:1 ratio. PMID- 11055337 TI - Synthesis and preliminary analysis of a P-glycoprotein-specific [3H]-benzophenone photoaffinity label based on (-)-stipiamide. AB - A benzophenone photoaffinity label 9 based on the polyene natural product (-) stipiamide has been constructed using a diaminoethane spacer and the radioactive agent [3H]-BZDC (N-succinimidyl p-benzoyl-(2,3-3H)-dehydrocinnamate). Photoaffinity experiments show specific binding to human P-glycoprotein (Pgp) in the presence of cis-flupentixol but not with cyclosporin A. PMID- 11055338 TI - Metal mediated protease inhibition: design and synthesis of inhibitors of the human cytomegalovirus (hCMV) protease. AB - A versatile synthetic route to a novel series of bis-imidazolemethanes designed to inhibit the hCMV protease has been developed and a series of potential metal binding inhibitors has been identified. In selectivity assays, the compounds were highly specific for CMV protease and showed no inhibition (IC50 > 100 microM) of other prototypical serine proteases such as trypsin, elastase, and chymotrypsin. Although the presence of free zinc ions was found to be an absolute requirement for the in vitro biological activity of this class of inhibitor, the potency of the inhibitors could not be improved beyond the micromolar level. PMID- 11055339 TI - Tetrahydroisoquinoline derivatives containing a benzenesulfonamide moiety as potent, selective human beta3 adrenergic receptor agonists. AB - Tetrahydroisoquinoline derivatives containing a 4-(hexylureido)benzenesulfonamide were examined as human beta3 adrenergic receptor (AR) agonists. Notably, 4,4 biphenyl derivative 9 was a 6 nM full agonist of the beta3 AR. Naphthyloxy compound 18 (beta3 EC50 = 78 nM) did not activate the beta1 and beta2 ARs at 10 microM, and showed >1000-fold selectivity over binding to the beta1 and beta2 ARs. PMID- 11055340 TI - DNA triplex structures are stabilized by the incorporation of 3'-endo blocked pyrimidine nucleosides in the Hoogsteen strand. AB - A short route to pyrimidine locked nucleosides has been developed for their incorporation in triplex forming oligonucleotides (TFO). Compared to oligonucleotides built with standard nucleosides, the modified TFOs containing 3' endo blocked residues formed, with their corresponding DNA duplexes, more stable triple helix systems, an effect which might be ascribed to the 3'-endo pucker of the modified nucleoside residues. PMID- 11055341 TI - Synthesis and enzymatic evaluation of a P1 arginine aminocoumarin substrate library for trypsin-like serine proteases. AB - A method for the solid-phase synthesis of P1 arginine containing peptides via attachment of the arginine side-chain guanidine group is described. This procedure is applied to the preparation of a tetrapeptide, P1 arginine aminocoumarin PS-SCL. This library was validated by using it to determine the P4 P2 specificity for thrombin and comparing the results to the known thrombin subsite specificity. This is the first reported example of a PS-SCL library containing a P1 arginine. PMID- 11055342 TI - N1-(Benzenesulfonyl)tryptamines as novel 5-HT6 antagonists. AB - N-Benzenesulfonyl-5-methoxy-N,N-dimethyltryptamine (BS/5-OMe DMT; 5) was shown to bind at human 5-HT6 serotonin receptors with high affinity (Ki = 2.3 nM) relative to serotonin (Ki = 78 nM). Structural variation failed to result in significantly enhanced affinity. BS/5-OMe DMT acts as an antagonist of 5-HT-stimulated adenylate cyclase (pA2 = 8.88 nM) and may represent the first member of a novel class of 5-HT6 antagonists. PMID- 11055343 TI - (E) -6-(1-alkyloxyiminoalkyl)-5,8-dimethoxy-1,4-naphthoquinones: synthesis, cytotoxic activity and antitumor activity. AB - All of 13 (E)-6-(1-alkyloxyiminomethyl)-5,8-dimethoxy-1,4-naphthoquinone derivatives synthesized showed high ED50 values, ranging from 0.1 to 0.3 microg/mL against L1210 cells. However, they were inactive on A549 cells. Nine compounds exhibited higher T/C (%) values (318-388%) than Adriamycin (T/C, 315%). PMID- 11055344 TI - Novel, potent and selective chimeric FXa inhibitors featuring hydrophobic P1 ketoamide moieties. AB - Judicious combination of P-region sequences of highly potent anticoagulant proteins including NAP5, NAP6, Ecotin, and Antistasin with SAR from small molecule FXa inhibitors led to a series of chimeric inhibitors of formula 1a-j. We report herein the design, synthesis, and biological activity of this novel family of FXa inhibitors that express both high in vitro potency and superb selectivity against related serine proteases. PMID- 11055345 TI - Synthesis of pyrrolo[2,1-c[1,4]benzodiazepines via reductive cyclization of omega azido carbonyl compounds by TMSI: an efficient preparation of antibiotic DC-81 and its dimers. AB - Azido carbonyl compounds on reaction with trimethylsilyl iodide (in situ prepared from TMSCl/NaI) led to the formation of diazepine imines in good yields under mild conditions. This methodology has been applied to the parent unsubstituted pyrrolobenzodiazepine, the natural product DC-81 and its dimers. PMID- 11055346 TI - Calpain inhibitors based on the quiescent affinity label concept: high rates of calpain inactivation with leaving groups derived from N-hydroxy peptide coupling reagents. AB - A series of irreversible inhibitors of recombinant calpain has been synthesized and their rates of inactivation have been evaluated against calpain and cathepsin B, respectively. The design of the inhibitors was based on the quiescent affinity label concept. By choosing the appropriate affinity group and by employing leaving groups derived from N-hydroxy coupling reagents, good inhibitors of calpain with high rates of inactivation have been identified. However, poor aqueous stability limits their therapeutic utility. PMID- 11055347 TI - Synthesis of derivatives of NK109, 7-OH benzo[c]phenanthridine alkaloid, and evaluation of their cytotoxicities and reduction-resistant properties. AB - The N5-C6 double bond of NK109 (an antitumor benzo[c]phenanthridine alkaloid) is easily reduced under biological environment. To suppress the inactivation caused by reduction, we synthesized 5-, 6-, and 8-substituted NK109. 5-Substituted derivatives (4a-c) were reduced more easily than NK109. 6-Substituted ones (10a f) inhibited biological reduction, but showed weak cytotoxic activity. 8-O Substituted ones (13a-h), especially 8-O-hydroxyethyl NK109 (13d), suppressed biological reduction and exhibited strong cytotoxic activity. PMID- 11055348 TI - Design and synthesis of a novel water soluble benzotetrazepinone. AB - In order to confer water solubility to the benzotetrazepinone ring system, the synthesis of 12 was undertaken. The design and synthesis of 12 were based upon previously established structural requirements for the stability of the 1,2,3,5 tetrazepin-4-one ring system. Tetrazepinone 12 was extremely water soluble and was 10-fold more potent than its imidazo-1,2,3,5-tetrazin-4-one counterpart 1a, against the human MCF-7 breast cancer cell line. PMID- 11055349 TI - The design and synthesis of novel NK1/NK2 dual antagonists. AB - Functional probing of the backbone of the Sanofi NK2 antagonist SR 48968 has resulted in the discovery of two new classes of NK1/NK2 dual antagonists: the diamine class and the oxime class. The addition of the amino or the oxime functional group results in the reversal of the stereochemical preference of the NK2 receptor. PMID- 11055350 TI - Synthesis and NK1/NK2 receptor activity of substituted-4(Z)-(methoxyimino)pentyl 1-piperazines. AB - A series of 5-[(3,5-bis(trifluoromethyl)phenyl)methoxy]-3-(3,4-dichlorophenyl) 4(Z)- (methoxyimino)pentyl-1-piperazines was prepared and their affinity for the NK1 and NK2 receptors investigated. Compounds 7f, 10o, 10r, and 10s were found to be our most potent inhibitors. PMID- 11055351 TI - Structure-based design and synthesis of phosphinate isosteres of phosphotyrosine for incorporation in Grb2-SH2 domain inhibitors. Part 1. AB - Based on X-ray crystal structure information, mono charged phosphinate isosteres of phosphotyrosine have been designed and incorporated in a short inhibitory peptide sequence of the Grb2-SH2 domain. The resulting compounds, by exploiting additional interactions, inhibit binding to the Grb2-SH2 domain as potently as the corresponding doubly charged (phosphonomethyl)phenylalanine analogue. PMID- 11055352 TI - Structure-based design and synthesis of phosphinate isosteres of phosphotyrosine for incorporation in Grb2-SH2 domain inhibitors. Part 2. AB - A series of novel phosphinates, derived from 4-phosphonomethylphenylalanine, are described as isosteres of phosphotyrosine. Benzyl (or alkyl) phosphinomethylphenylalanine derivatives were prepared by alkylation of an amino acid P-H phosphinate. PMID- 11055353 TI - 1,2-Disubstituted indole, azaindole and benzimidazole derivatives possessing amine moiety: a novel series of thrombin inhibitors. AB - A novel series of 1,2-disubstituted indole, azaindole and benzimidazole derivatives possessing an amine moiety was identified as thrombin inhibitors. An indole with basic diamine moieties (12a) was the most potent thrombin inhibitor in the series with Kass= 197 x 10(6) L/mol. PMID- 11055354 TI - Highly sensitive and rapid detection of antibody catalysis by luminescent bacteria. AB - A highly sensitive, inexpensive, and facile bioluminescent assay for the detection of catalytic antibodies has been developed. This assay may be used for the early detection of antibody catalysis. The efficiency of this technique was exemplified by the use of the luminescent bacterium VhM42 for monitoring an antibody-catalyzed retroaldol fragmentation reaction with aldolase antibodies 38C2 and 24H6. PMID- 11055355 TI - Dibasic inhibitors of human mast cell tryptase. Part 1: synthesis and optimization of a novel class of inhibitors. AB - The synthesis and optimization of a novel class of reversible and active-site directed dibasic inhibitors of human mast cell tryptase are described. The compounds were shown to be both remarkably potent and selective for tryptase with Ki values for optimized inhibitors in the picomolar range. PMID- 11055356 TI - Dibasic inhibitors of human mast cell tryptase. Part 2: structure-activity relationships and requirements for potent activity. AB - Detailed structure activity relationships (SARs) for a series of dibasic human tryptase inhibitors are presented. The structural requirements for potent inhibitory activity are remarkably broad with a range of core template modifications being well tolerated. Optimized inhibitors demonstrate potent anti asthmatic activity in a sheep model of allergic asthma. APC-2059, a dibasic tryptase inhibitor with subnanomolar activity, has been advanced to phase II clinical trials for the treatment of both psoriasis and ulcerative colitis. PMID- 11055357 TI - Optimising inhibitors of trypanothione reductase using solid-phase chemistry. AB - A series of inhibitors of the enzyme trypanothione reductase has been identified using directed solid-phase chemistry. The compounds were based on a series of polyamine scaffolds and used the natural product kukoamine A as the lead structure. A compound with a Ki of 76 nM was identified, although somewhat surprisingly the compound appeared to be noncompetitive in nature. PMID- 11055358 TI - Chemical modification of nodulisporic acid A: preliminary structure-activity relationships. AB - Medicinal chemistry efforts were initiated to identify the key constituents of the nodulisporic acid A (1) pharmacophore that are integral to its potent insecticidal activity. New semisynthetic derivatives delineated 1 into 'permissive' and 'nonpermissive' regions and led to the discovery of new nodulisporamides with significantly improved flea efficacy. PMID- 11055359 TI - Novel heterocycles as selective alpha1-adrenergic receptor antagonists. AB - A novel series of aryl piperazine substituted heterocycles has been synthesized and identified as antagonists of the alpha1a-adrenergic receptor (alpha1a-AR), which has been implicated in benign prostatic hyperplasia (BPH). These compounds selectively inhibit binding to the alpha1a-AR with K(i)s as low as 2.1 nM. PMID- 11055360 TI - Analogues and derivatives of ciproxifan, a novel prototype for generating potent histamine H3-receptor antagonists. AB - Novel derivatives of the highly potent and selective histamine H3-receptor antagonist ciproxifan (3) with different chain lengths as well as with structural variants of the cyclopropyl ketone moiety have been prepared and screened for their antagonist H3-receptor potencies in vitro and in vivo. Some derivatives (2, 6-8, 12) containing other functionalities were effective in vitro in the same (sub)nanomolar concentration range and in vivo in a remarkably low oral dose. PMID- 11055361 TI - Synthesis and estrogen receptor binding affinities of novel pyrrolo[2,1,5 cd]indolizine derivatives. AB - A series of pyrrolo[2,1,5-cd]indolizine derivatives has been synthesized and evaluated as ligands for the estrogen receptor. Properly substituted mono- and di hydroxy derivatives showed binding in the low nanomolar range in accordance with their structural resemblance to estrogen. PMID- 11055362 TI - Synthesis and biological activity of a bivalent nucleotide inhibitor of ribonucleotide reductase. AB - A novel nucleotide inhibitor (ADP-S-HBES-S-dGTP) of mouse ribonucleotide reductase was designed to span the active site and the allosteric specificity site of the enzyme. The inhibitor contains ADP and dGTP moieties which are linked by 1,6-hexane-(bis-ethylenesulfone), and has a Ki value of 12 microM. PMID- 11055363 TI - Least squares estimation of variance components for linkage. AB - We develop least squares (LS) procedures for variance components estimation in genetic linkage studies. The LS procedure is expressed by simple expressions, and does not require inversion of large matrices. Simulations comparing LS with maximum likelihood (ML) procedures for normal data show that both yield unbiased estimators, but the efficiency of the LS procedure was less than 50% of the ML procedure. For bivariate normal data, the efficiency of the LS procedure relative to the ML method was better, generally over 60%. For skewed data, the LS method was markedly more efficient than ML for parameter estimation. The LS method was computationally rapid, over 4,000 times faster than ML estimation for bivariate data. Because ML estimation is time consuming, LS methods are suggested for initial interval mapping with multivariate data. PMID- 11055364 TI - Robust LOD scores for variance component-based linkage analysis. AB - The variance component method is now widely used for linkage analysis of quantitative traits. Although this approach offers many advantages, the importance of the underlying assumption of multivariate normality of the trait distribution within pedigrees has not been studied extensively. Simulation studies have shown that traits with leptokurtic distributions yield linkage test statistics that exhibit excessive Type I error when analyzed naively. We derive analytical formulae relating the deviation from the expected asymptotic distribution of the lod score to the kurtosis and total heritability of the quantitative trait. A simple correction constant yields a robust lod score for any deviation from normality and for any pedigree structure, and effectively eliminates the problem of inflated Type I error due to misspecification of the underlying probability model in variance component-based linkage analysis. PMID- 11055365 TI - Sampling among haplotype resolutions in a coalescent-based genealogy sampler. AB - Analysis of the coalescent structure of a population may provide information useful in mapping disease loci. Current coalescent-based genealogy samplers require haplotyped data, but haplotypes are not always available, and it is not practical to sum over all haplotype assignments for large data sets. We describe a method of adding haplotype re-evaluation to the sampler, so that it samples not only among genealogies explaining a given haplotype configuration, but also among different haplotype configurations. Several different haplotype-rearrangement strategies are considered, but the simplest-inverting the phase of a single site in a single individual-appears to be the most successful. The straightforward haplotype sampler does not mix well; heating approaches can greatly improve its performance. PMID- 11055366 TI - Variance-Components QTL linkage analysis of selected and non-normal samples: conditioning on trait values. AB - Standard variance-components quantitative trait loci (QTL) linkage analysis can produce an elevated rate of type 1 errors when applied to selected samples and non-normal data. Here we describe an adjustment of the log-likelihood function based on conditioning on trait values. This leads to a likelihood ratio test that is valid in selected samples and non-normal data, and equal in power to alternative methods for analyzing selected samples that require knowledge of the ascertainment procedure or the trait values of non-selected individuals. PMID- 11055367 TI - An analysis of strategies for discovery of single-nucleotide polymorphisms. AB - Strategies for the discovery of single-nucleotide polymorphisms (SNPs) can be characterized by the number of individuals in the discovery sample, and by the minimal required number of observations of each allele. We examine the effect of different strategies on two key properties of the resulting SNP collection: (1) the probability that a SNP with a given population allele frequency is detected; and (2) the allele-frequency distribution of the discovered SNPs. We show that strategies that accept all polymorphic sites lead to collections with a high fraction of SNPs with rare minor alleles, particularly in expanded populations. Such SNPs have a low probability of replication in a second sample. We discuss how to tailor a discovery strategy to the desired properties of a SNP collection. PMID- 11055368 TI - Implementing a unified approach to family-based tests of association. AB - We describe a broad class of family-based association tests that are adjusted for admixture; use either dichotomous or measured phenotypes; accommodate phenotype unknown subjects; use nuclear families, sibships or a combination of the two, permit multiple nuclear families from a single pedigree; incorporate di- or multi allelic marker data; allow additive, dominant or recessive models; and permit adjustment for covariates and gene-by-environment interactions. The test statistic is basically the covariance between a user-specified function of the genotype and a user-specified function of the trait. The distribution of the statistic is computed using the appropriate conditional distribution of offspring genotypes that adjusts for admixture. PMID- 11055369 TI - A general test of association for complex diseases with variable age of onset. AB - Analysis of age of onset is a key factor in linkage and association studies of some complex genetic traits. Recent methodological developments are mainly concentrated on binary or quantitative traits, in which age of onset information is ignored. We propose a linkage disequilibrium-based Cox model and a robust score test for testing association between a marker and a disease with variable age of onset. The proposed model is semi-parametric, with an unspecified baseline hazard function. Simulation results indicate that the proposed methods have correct error rates and good statistical power, even in the presence of population admixture. This approach offers a solution to the problem of testing for marker associations when there is a variable age of onset. PMID- 11055370 TI - Bayesian linkage and segregation analysis: factoring the problem. AB - Complex segregation analysis and linkage methods are mathematical techniques for the genetic dissection of complex diseases. They are used to delineate complex modes of familial transmission and to localize putative disease susceptibility loci to specific chromosomal locations. The computational problem of Bayesian linkage and segregation analysis is one of integration in high-dimensional spaces. In this paper, three available techniques for Bayesian linkage and segregation analysis are discussed: Markov Chain Monte Carlo (MCMC), importance sampling, and exact calculation. The contribution of each to the overall integration will be explicitly discussed. PMID- 11055371 TI - Clustering methods applied to allele sharing data. AB - Here we focus on using clustering methods to disentangle the interacting factors that lead to the presentation of complex diseases. Relative pairs are placed in discrete subgroups, or classes, based upon their pattern of allele sharing at a sequence of markers and on concomitant risk factors. The relationship between the locus information and the affectation status of the relative pairs within each subgroup then can be assessed. Cluster analysis (CLA) and latent class analysis (LCA) were applied to sibling allele sharing data from GAW11 simulated data, and to an existing Alzheimer's disease (AD) dataset. Both methods were able to identify markers linked to all 3 disease loci in the GAW11 data. LCA and CLA also replicated regions of chromosomes identified in an analysis of the AD data using affected-sib-pair methods. These analyses indicate that classification tools may be useful for detecting susceptibility genes for complex traits. PMID- 11055372 TI - Zero-recombinant haplotyping: applications to fine mapping using SNPs. AB - As the number of single nucleotide polymorphisms (SNPs) available for genetic analysis increases, researchers will be saturating smaller and smaller regions of the genome with these biallelic markers in an effort to fine map complex diseases. An important tool in this fine-mapping effort is haplotyping. Algorithms are presented that find all possible haplotype configurations of the pedigree data under the assumption that there are no recombinants between the markers. These configurations can be used to estimate the haplotype frequencies, and identify the most common haplotypes in the data. These algorithms have been implemented into a software program (ZAPLO), and were tested on a published data set. PMID- 11055373 TI - Methods for multipoint disease mapping using linkage disequilibrium. AB - A disease-associated mutation arises on a single chromosome such that alleles at linked markers are initially in complete linkage disequilibrium (LD) with the mutation. LD can be used as a tool for high-resolution mapping of the position of a disease mutation relative to a set of linked marker loci. When more than two linked marker loci are considered, developing a maximum likelihood approach is a challenging mathematical problem. To reduce the complexity, approximate and composite likelihood (CL) methods have been developed for multipoint LD mapping that use simplified models of population history, or of recombination, that ignore some of the statistical dependence among disease chromosomes and among marker loci. We describe the relationship among several composite likelihood methods for multipoint LD mapping, and suggest an alternative CL method that takes better account of the statistical dependence among marker loci. PMID- 11055374 TI - Robust transmission regression models for linkage and association. AB - We present a general regression model that accounts for both linkage and linkage disequilibrium (LD) when analyzing nuclear family data. The method does not require LD to exist in order to evaluate linkage, but if LD does exist, the power to detect linkage can increase due to improved information on linkage phase. The proposed method is general, allowing for a variety of traits (e.g., binary affection status, categorical and quantitative phenotypes), affecteds only analyses, and covariates. Covariates can be useful to assess heterogeneity of linkage and LD, as well as gene-environment interactions. Other advantages of our methods are that: LD parameters are not defined without linkage, so that population stratification cannot bias the analyses; a combined test for linkage and LD can be used to test for linkage; given the existence of linkage, an adjusted LD test useful for fine-mapping can be constructed; covariate effects can be flexibly modeled; and families containing a single child and families containing multiple offspring can be combined for a single analysis (capitalizing on the LD information provided by single-child families and the combined linkage and LD information provided by multiple offspring). The basic features of the regression model are presented, as well as discussions of potential applications and critical statistical issues. PMID- 11055375 TI - Power of a score test for quantitative trait linkage analysis of relative pairs. AB - The score test of Dudoit and Speed [(2000) Biostatistics 1:1-26] to detect linkage between a trait locus and a marker locus, using identity by descent data on sib pairs, is extended to other types of relative pairs (grandparent/grandchild, avuncular, and half-sib relationships). The test is based on the likelihood of the recombination fraction theta between trait and marker loci, conditional on phenotypes of the relatives. We present results of simulation studies characterizing power and robustness properties of this linkage score test, and compare the power of the score test to that of the classical and modified Haseman-Elston tests. The score test has considerable power, particularly under sampling schemes where selection is on double probands. Use of a generic additive model [Goldstein et al., submitted] with allele frequency p = 0.2, heritability H = 0.3, and a moderate residual correlation of rho = 0.2 resulted in a very good overall performance across a wide range of trait generating models. PMID- 11055376 TI - Comparison of allele-sharing statistics for general pedigrees. AB - The popularity of "non-parametric" linkage analysis using moderate and large sized pedigrees has increased in the last few years. Only a few of the existing allele-sharing statistics are applicable to general pedigrees, and even fewer are in common use. Little research has been done to compare the power of these statistics. We present some preliminary investigations into the power of the most popular general-pedigree allele-sharing statistics. We show that there are indeed non-trivial differences among the statistics, and that further studies are warranted. In particular, pedigree structure and the degree of recessiveness of the trait appear to be critical factors in determining which statistics are most powerful. PMID- 11055377 TI - Linkage disequilibrium mapping in populations of variable size using the decay of haplotype sharing and a stepwise-mutation model. AB - Linkage-disequilibrium (LD) mapping is a powerful tool for fine-mapping disease genes. Recently, McPeek and Strahs [(1999) Am J Hum Genet 65:858-875] proposed a multilocus model for LD mapping based on the decay of haplotype sharing. Here we extend their approach in two ways. First, instead of assuming each marker allele has an equal chance to mutate to one of the other marker alleles, we use the stepwise-mutation model to describe the mutation process for microsatellite markers. Second, in addition to the independence model and the constant population size model they considered, we model the dependence among observed haplotypes due to population structure by using a general conditional-coalescent model with variable population size. Through simulation studies, we study the effects of the stepwise-mutation model and variable population size on the estimates of disease gene location, mutation rate, and time to the most recent common ancestor of the sampled haplotypes. We then use this method to analyze progressive myoclonus epilepsy data. PMID- 11055378 TI - Erythropoietin: multiple physiological functions and regulation of biosynthesis. AB - Erythropoietin (Epo), which is produced by the kidney in the adult and by the liver in the fetus, increases red blood cells by supporting the survival of erythroid progenitor cells and stimulating their differentiation and proliferation via binding to Epo receptor (EpoR). The main signal in the control of Epo production is oxygen; hypoxia stimulates Epo production through activation of Epo gene transcription. Tremendous progress in our understanding of molecular mechanisms of Epo action on erythroid cells and regulation of the Epo production has been made by manipulation of cDNAs and genes of Epo and EpoR. Studies on hypoxic induction of Epo gene transcription led to the identification of hypoxia inducible factor (HIF-1), a transcriptional factor, that functions as a global regulator of hypoxic gene expression. Paracrine Epo/EpoR systems that are independent of the endocrine erythropoietic system (kidney/bone marrow) have been found in the central nervous system and uterus. Novel functions of Epo at these local sites and tissue-specific regulation of Epo production including a newly found potent regulator (estrogen) have been proposed. The tissue-specific regulation rationalizes the specific functions of Epo produced by individual tissues. PMID- 11055379 TI - Effects of germinated barley foodstuff on microflora and short chain fatty acid production in dextran sulfate sodium-induced colitis in rats. AB - Germinated barley foodstuff (GBF) administration has been previously reported to suppress dextran sulfate sodium (DSS)-induced experimental colitis. In this study, we investigated the roles of the intestinal microflora and short chain fatty acids (SCFAs) following administration of GBF in DSS-induced rat colitis. Sprague-Dawley rats were fed 3% (w/w of diet) DSS in GBF-diets for 5 days. The control rats were fed 3% DSS in cellulose-diets for 5 days. The administration of GBF effectively prevented bloody diarrhea and mucosal damage as compared to control rats. GBF significantly elevated fecal acetic acid and n-butyric acid levels. GBF tended to increase the number of eubacteria and that of bifidobacteria as compared to control rats. In addition, the number of enterobacteriaceae, the total number of aerobes and bacteroidaseae, were significantly lower in rats fed GBF than in the control group. It is suggested that the therapeutic effects of GBF for DSS-induced colitis depend mainly on increased SCFAs, which are accompanied by changes of composition of intestinal bacteria. PMID- 11055380 TI - Ascorbic acid stimulation of production of a highly branched ,beta-1,3-glucan by Aureobasidium pullulans K-1--oxalic acid, a metabolite of ascorbic acid as the stimulating substance. AB - The production of a highly branched beta-1,3-glucan by Aureobasidium pullulans K 1 in Czapek's medium has been found to be stimulated by ascorbic acid. When the culture supernatant, after removal of polysaccharide from the culture filtrate by ethanol precipitation, was concentrated, then added to a new medium and this strain was cultured in the medium, the polysaccharide production was stimulated the same as when L-ascorbic acid was added to the medium. The stimulating substance was partially purified from the supernatant, and was found to be oxalic acid; 0.03% oxalic acid was the most effective concentration for the stimulation of polysaccharide production. The stimulating substance, oxalic acid, was proved to be derived from ascorbic acid added to a medium in an experiment using L-[1 14C]ascorbic acid. We suggest that oxalic acid generated from the metabolism of ascorbic acid in cells of Aureobasidium pullulans K-1 participated in the stimulation of the polysaccharide production by ascorbic acid. PMID- 11055381 TI - Production of betacyanins by a cell suspension culture of table beet (Beta vulgaris L.). AB - A cell suspension culture of table beet (Beta vulgaris L.) was established for efficient betacyanin production from violet callus induced from the hypocotyls of aseptic seedlings. This suspension culture produced large amounts of betacyanins. The betacyanin content increased with increasing cell growth during the log phase. Reducing the total nitrogen concentration (30 mM) and modifying the ratio of ammonium to nitrate (1:14) resulted in an increased betacyanin content. Supplementation of Fe2+ to the LS medium also promoted betacyanin production. The maximal betacyanin yield was achieved with a 2 mM Fe2+ concentration. Combining these conditions, we established a revised LS medium to improve betacyanin productivity (250 mg/l for a 14-day culture). PMID- 11055382 TI - Flavonoids inhibit cell growth and induce apoptosis in B16 melanoma 4A5 cells. AB - We investigated the growth inhibitory activity of several flavonoids, including apigenin, luteolin, kaempherol, quercetin, butein, isoliquiritigenin, naringenin, genistein, and daizein against B16 mouse melanoma 4A5 cells. Isoliquiritigenin and butein, belonging to the chalcone group, markedly suppressed the growth of B16 melanoma cells and induced cell death. The other flavonoids tested showed little growth inhibitory activity and scarcely caused cell death. In cells treated with isoliquiritigenin or butein, condensation of nuclei and fragmentation of nuclear DNA, which are typical phenomena of apoptosis, were observed by Hoechst 33258 staining and by agarose gel electrophoresis of DNA. Flowcytometric analysis showed that isoliquiritigenin and butein increased the proportion of hypodiploid cells in the population of B16 melanoma cells. These results demonstrate that isoliquiritigenin and butein inhibit cell proliferation and induce apoptosis in B16 melanoma cells. Extracellular glucose decreased the proportion of hypodiploid cells that appeared as a result of isoliquiritigenin treatment. p53 was not detected in cells treated with either of these chalcones, however, protein of the Bcl-2 family were detected. The level of expression of Bax in cells treated with either of these chalcones was markedly elevated and the level of Bcl-XL decreased slightly. Isoliquiritigenin did not affect Bcl-2 expression, but butein down-regulated Bcl-2 expression. From these results, it seems that the pathway by which the chalcones induce apoptosis may be independent of p53 and dependent on proteins of the Bcl-2 family. It was supposed that isoliquiritigenin induces apoptosis in B16 cells by a mechanism involving inhibition of glucose transmembrane transport and promotion of Bax expression. On the other hand, it was suggested that butein induces apoptosis via down regulation of Bcl-2 expression and promotion of Bax expression. This mechanism differs from the isoliquiritigenin induction pathway. PMID- 11055383 TI - Synthesis of alpha-D-glucosylglycerol by alpha-glucosidase and some of its characteristics. AB - It has been found that alpha-D-glucosylglycerol (GG) is contained in such traditional Japanese foods brewed by using koji as sake, miso and mirin, and that GG is formed by transglucosylation to glycerol that is produced by yeast with alpha-glucosidase (EC 3.2.1.20) from koji in the sake mash. GG has also been found to consist of three components, 2-O-alpha-D-glucosylglycerol (GG-II), (2R) 1-O-alpha-D-glucosylglycerol (R-GG-I) and (2S)-1-O-alpha-D-glucosylglycerol (S-GG I). GG was synthesized from a mixture of maltose and glycerol by the batch method, using alpha-glucosidase (transglucosidase L-AMANO). alpha-Glucosidase seemed to be so stable that the amount of GG increased about 5-fold compared with that in the first reaction by the daily addition of maltose for 10 d. Syrupy GG obtained was found to have the following characteristics: about 0.55-fold sweetness compared with sucrose, high thermo-stability, low heat-colorability, low Maillard reactivity, low hygroscopicity, high water-holding capacity, non cariogenicity and low digestibility. PMID- 11055384 TI - Interaction of gum arabic, maltodextrin and pullulan with lipids in emulsions. AB - The interaction of gum arabic, maltodextrin and pullulan with lipids in emulsion systems was investigated. Interfacial tension and interfacial viscosity measurements revealed that only gum arabic could adsorb and form a viscoelastic film at the oil-water interface. Good emulsifying activity was demonstrated for gum arabic, whereas fine emulsions could not be produced from the other polysaccharide solutions and oil. Frequency-dependent increases in the storage and loss moduli were observed for all the polysaccharide solutions. Such rheological behavior did not substantially change when maltodextrin and pullulan were mixed with oil to form emulsions. However, the frequency-dependence of the dynamic moduli disappeared in the gum arabic-stabilized emulsion, suggesting the formation of a network structure in which oil droplets could form junctions with gum arabic chains. The results on the inhibition of lipid oxidation by polysaccharides suggest that gum arabic protected lipids from the attack of lipoxygenase and free radicals by adsorbing at the oil droplet surface. PMID- 11055385 TI - Preparative 2'-reduction of ATP catalyzed by ribonucleotide reductase purified by liquid-liquid extraction. AB - Recombinant Lactobacillus leichmannii ribonucleosidetriphosphate reductase (RTPR, E.C.1.17.4.2) constitutively expressed by E. coli HB101 pSQUIRE has been purified from sonicated cell material in a one-step procedure by PEG 4000 (16% (w/w))/phosphate (7% (w/w)) liquid-liquid extraction. A high yield of 75.1% RTPR in the top phase and a partitioning of 8.5:1 between total RTPR activity in top and bottom phase were obtained in this preparative system. The RTPR-containing top phase was used to reduce ATP in the 2'-position on a gram scale with high final conversion and yield proving the ribonucleotide reductase approach feasible for the preparative synthesis of 2'-deoxyribonucleotides. High concentrations of sodium acetate in the reaction served to substitute for allosteric effectors of RTPR. 1,4-Dithio-DL-threitol was used as an artificial reducing agent for RTPR. PMID- 11055386 TI - Absolute configuration of a ceramide with a novel branched-chain fatty acid isolated from the epiphytic dinoflagellate, Coolia monotis. AB - The absolute configuration of the chiral center at the C15 position of a novel branched-chain fatty acid derived from a new ceramide isolated from the epiphytic dinoflagellate Coolia monotis was determined to be of R from by reversed-phase HPLC after cleavage to 12-methylpentadecanoic acid and subsequent conversion with the chiral fluorescent reagent, (1R,2R)-2-(2,3 anthracenedicarboximido)cyclohexanol. PMID- 11055387 TI - Structural analysis of free N-glycans occurring in soybean seedlings and dry seeds. AB - The structures of unconjugated or free N-glycans in stems of soybean seedlings and dry seeds have been identified. The free N-glycans were extracted from the stems of seedlings or defatted dry seeds. After desalting by two kinds of ion exchange chromatography and a gel filtration, the free N-glycans were coupled with 2-aminopyridine. The resulting fluorescence-labeled (PA-) N-glycans were purified by gel filtration, Con A affinity chromatography, reverse-phase HPLC, and size-fractionation HPLC. The structures of the PA-sugar chains purified were analyzed by the combination of two-dimensional sugar chain mapping, jack bean alpha-mannosidase digestion, alpha-1,2-mannosidase digestions, partial acetolysis, and ESI-MS/MS. The free N-glycan structures found showed that two categories of free N-glycans occur in the stems of soybean seedlings. One is a high-mannose type structure having one GlcNAc residue at the reducing end (Man 9 approximately 5 GlcNAc1, 93%), that would be derived by endo-GM (Kimura, Y. et al., Biochim. Biophys. Acta, 1381, 27-36 (1998)). The other small component is a xylose-containing type one having two GlcNAc residues at the reducing end (Man3Xyl1GlcNAc2, 7%), which would be derived by PNGase-GM (Kimura, Y. and Ohno, A., Biosci. Biotechnol. Biochem., 62, 412-418 (1998)). The detailed structural analysis of free glycans showed that high-mannose type free N-glycans (Man 9 approximately 5 GlcNAc1) in the soybean seedlings have a common core structural unit; Manalpha1-6(Man1-3)Manalpha1-6(Manalpha1-3)Ma nbeta1-4GlcNAc. Comparing the amount of free N-glycans in the seedling stems and dry seeds, the amount in the stems of seedlings was much higher than that in the dry seeds; approximately 700 pmol per one stem, 8 pmol in one dry seed. This fact suggested that free N glycans in soybean seedlings could be produced by two kinds of N-glycan releasing enzymes during germination or seedling-development. PMID- 11055388 TI - Epitope analysis and primary structures of variable regions of anti-human FcepsilonRI monoclonal antibodies, and expression of the chimeric antibodies fused with human constant regions. AB - The structural analysis of monoclonal antibodies (mAbs) against the alpha subunit of the high affinity IgE receptor (FcepsilonRIalpha) is an alternative approach to obtaining information for the design of inhibitors that will block complementary interaction between IgE and FcepsilonRIalpha and to analyzing the various biological effects induced by anti-FcepsilonRIalpha autoantibodies in chronic urticaria. In this study, epitopes for mouse anti-human FcepsilonRIalpha mAbs and primary structures of variable regions of the mAbs were analyzed. Three mAbs inhibitory for IgE-binding reacted to the deletion mutants of FcepsilonRIalpha containing the whole second immunoglobulin-like domain as well as IgE did. On the other hand, two uninhibitory mAbs reacted to those containing the whole first immunoglobulin-like domain. The cDNAs for variable regions of the five mAbs were cloned and sequenced. Two inhibitory mouse/human chimeric antibodies were expressed in COS7 cells and bound to Chinese hamster ovary transfectant cells expressing FcepsilonRI (CHO/alphabetagamma), and these inhibited the binding of IgE to CHO/alphabetagamma cells. PMID- 11055389 TI - Preventive effect of lactobacillus delbrueckii subsp. bulgaricus on the oxidation of LDL. AB - Lactobacillus delbrueckii subsp. bulgaricus 2038 was examined for its activity to prevent the oxidation of the erythrocyte membrane in vitro, and the oxidation of LDL in vivo. Strain 2038 produced radical scavengers that reacted with 1,1 diphenyl-2-picrylhydrazl (DPPH) during cultivation. Moreover, the ethereal extract from the supernatant of the culture prevented the oxidation of the erythrocyte membrane in vitro. As an in vivo study, male F344 rats were fed on diets containing 20% fresh soybean oil (or 13% oxidized oil and 7% fresh oil) with 10% freeze-dried powder of the 2038 culture (or with skim milk powder) for 4 weeks. The level of thiobarbituric acid-reactive substances was lower in the low density lipoproteins (per milligram of cholesterol) from rats fed on the oxidized oil with freeze-dried powder of the 2038 culture than without it. The level of vitamin E in the plasma was higher in the rats fed on the oxidized oil with the freeze-dried powder than without it. PMID- 11055390 TI - Characteristics of serine acetyltransferase from Escherichia coli deleting different lengths of amino acid residues from the C-terminus. AB - Some properties of serine acetyltransferases (SATs) from Escherichia coli, deleting 10-25 amino acid residues from the C-terminus (SATdeltaC10-deltaC25) were investigated. The specific activity depended only slightly on the length of the C-terminal region deleted. Although the sensitivity of SATdeltaC10 to inhibition by L-cysteine was similar to that for the wild-type SAT, it became less with further increases in the length of the amino acid residues deleted. SATdeltaC10 was inactivated on cooling to 0 degrees C and dissociated into dimers or trimers in the same manner as the wild-type SAT, but Met-256-le mutant SAT as well as SATdeltaC14, SATdeltaC20, and SATdeltaC25 were stable. Since SATdeltaC10, SATdeltaC14, and SATdeltaC25 did not form a complex with O-acetylserine sulfhydrylase-A (OASS-A) in a way similar to SATdeltaC20, it was indicated that 10 amino acid residues or fewer from the C-terminus of the wild-type SAT are responsible for the complex formation with OASS-A. The C-terminal peptide of the 10 amino acid residues interacted competitively with OASS-A with respect to OAS although its affinity was much lower than that for the wild-type SAT. PMID- 11055391 TI - Safety assessment of rice genetically modified with soybean glycinin by feeding studies on rats. AB - Feeding studies on rice genetically modified with soybean glycinin were performed on rats for four weeks. The rats were divided into three groups, each being fed on (I) only a commercial diet, (II) this diet plus control rice and (III) this diet plus rice genetically modified with glycinin. The rats were fed with 10 g/kg weight of rice every day by oral administration. During the test period, the rats in every group grew well without marked differences in appearance, food intake, body weight, or cumulative body weight gain. There were also no significant differences in the blood count, blood composition or internal organ weights among the rats. Necropsy at the end of the experiment indicated neither pathological symptoms nor histopathological abnormalities in the liver and kidney. Judging from these results, the rice genetically modified with glycinin is considered to have been essentially the same in nutritional and biochemical characteristics as the control rice. PMID- 11055392 TI - Meat allergy: investigation of potential allergenic proteins in beef. AB - The potential allergenic proteins in beef were investigated. The sera of ten beef allergic patients suffering from atopic dermatitis and having a positive RAST score to beef, aged 3-18 years, were obtained from Yoshida Hospital in Japan, and five non-allergic individuals were subjected to this study. The sera of the ten patients reacted strongly to a beef extract, but not to pork and chicken extracts by both ELISA and immunoblotting. The sera of the five control subjects did not react to any of these meat extracts. Three bands having molecular masses of approximately 200 kDa, approximately 67 kDa and approximately 60 kDa were observed by immunoblotting after SDS-PAGE. Two fractions of the beef extract from a Sephadex-gel (G-200) filtration column strongly reacted with the sera of the beef-allergic patients by ELISA and immunoblotting: one fraction had the approximately 67 kDa component and the other had the approximately 200 kDa and approximately 60 kDa components. One of them (approximately 67 kDa) was confirmed to be bovine serum albumin (BSA) by an analysis of the N-terminal amino acid sequence. We could not identify the others by sequencing, but the approximately 200 kDa and approximately 60 kDa components were presumed to be glycoproteins. Bovine gamma (BGG:globulin M.W. approximately 160 kDa) is a glycoprotein and has several subunits. The beef-allergic patients showed strong reactivity to the approximately 200 kDa and approximately 60 kDa components of pure BGG by immunoblotting. Inhibition-ELISA showed that pure BGG preparations strongly inhibited the binding of sera from the beef-allergic patients to the beef extract. These results suggest that the approximately 200 kDa, approximately 67 kDa and approximately 60 kDa components in the beef extract had strong allergenicity: approximately 67 kDa was BSA, and approximately 200 kDa and approximately 60 kDa were presumably aggregated BGG and it's heavy chain, respectively. PMID- 11055393 TI - Purification and characterization of chitosanase and Exo-beta-D-glucosaminidase from a Koji mold, Aspergillus oryzae IAM2660. AB - Chitosan-degrading activity was detected in the culture fluid of Aspergillus oryzae, A. sojae, and A. flavus among various fungal strains belonging to the genus Aspergillus. One of the strong producers, A. oryzae IAM2660 had a higher level of chitosanolytic activity when N-acetylglucosamine (GlcNAc) was used as a carbon source. Two chitosanolytic enzymes, 40 kDa and 135 kDa in molecular masses, were purified from the culture fluid of A. oryzae IAM2660. Viscosimetric assay and an analysis of reaction products by thin-layer chromatography clearly indicated the endo- and exo-type cleavage manner for the 40-kDa and 135-kDa enzymes, respectively. The 40-kDa enzyme, designated chitosanase, catalyzed a hydrolysis of glucosamine (GlcN) oligomers larger than pentamer, glycol chitosan, and chitosan with a low degree of acetylation (0-30%). The 135-kDa exo-beta-D glucosaminidase,enzyme,named released a single GlcN residue from the GlcN oligomers and chitosan, but did not release GlcNAc residues from either GlcNAc oligomer or colloidal chitin. PMID- 11055394 TI - Cloning and expression of a novel murine anti-human Fas antibody. AB - Agonistic anti-human Fas antibodies that can induce apoptosis are thought to have therapeutic effects for various diseases resulting from an abnormality of the Fas/FasL system. However, some anti-Fas antibodies show toxicity, and it is difficult to investigate their therapeutic and toxicological effect using animals because of their species specificity. We previously obtained a murine anti-human Fas mAb, HFE7A. HFE7A reacted with both human and murine Fas, and mitigated lymphadenopathy without any sign of hepatotoxicity in MRLgld/gld mice. It is suggested that humanized HFE7A would be a therapeutic treatment for various diseases resulting from an abnormality of the Fas/FasL system. Here we isolated the cDNAs that code for the heavy and light chains of HFE7A and identified the corresponding nucleotide sequences. The recombinant HFE7A was indistinguishable in binding and apoptosis-inducing activity to that from a hybridoma cell line. These data provide essential information for the humanization and clinical application of the humanized HFE7A. PMID- 11055395 TI - Kurosu, a traditional vinegar produced from unpolished rice, suppresses lipid peroxidation in vitro and in mouse skin. AB - The in vitro antioxidative activities of various kinds of vinegar were investigated by using a linoleic acid autoxidation model detected by the thiobarbituric acid (TBA) method and the 1,1-diphenyl-2-picrylhydrazyl radical system. An ethyl acetate extract of Kurosu (EK), a vinegar made from unpolished rice, exhibited the highest antioxidative activity in both systems. EK (5 mg) inhibited 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced edema formation (14%) and myeloperoxidase activity (52%, P< 0.01) in female ICR mouse skin. Furthermore, EK significantly suppressed double TPA application-induced H2O2 generation (53%, P< 0.01) and lipid peroxidation determined by the TBA-reacting substance level (95 %, P< 0.01). In a two-stage carcinogenesis experiment with dimethylbenz[a]anthracene/TPA, EK significantly reduced the number of tumors per mouse by 36% (P<0.05) at 15 weeks after promotion. These results suggest that the antitumor-promoting effect may be partially due to the antioxidative properties of EK such as the decomposition of free radicals and interference with free radical-generating leukocytes. PMID- 11055396 TI - Synthesis of 2-C-methyl-D-erythritol and 2-C-methyl-L-threitol; determination of the absolute configuration of 2-C-methyl-1,2,3,4-butanetetrol isolated. AB - 2-C-Methyl-D-erythritol (A) and 2-C-methyl-L-threitol (B) were respectively synthesized from D-glucose and D-galactose. The 2-C-methyl-1,2,3,4-butanetetrol compound (C) recently isolated from Phlox sublata L was confirmed to be A by comparing the CD and 1H-NMR spectra of its tri-O-benzoate with those of A and B. PMID- 11055397 TI - Thermally induced disintegration of the Bacillus stearothermophilus dihydrolipoamide dehydrogenase. AB - Upon heat treatment of the pyruvate dehydrogenase complex from Bacillus stearothermophilus, the most thermostable component is a dihydrolipoamide dehydrogenase (E3c). To understand this stability, the thermal disintegration of E3 dissociated from the complex (E3d) was examined, comparing with that of E3c. Judging from residual activity and inactivation rate, E3d was less thermostable than E3c; E3d and E3c lost half of their original activities upon incubations for 30 min at 79 degrees C and 90 degrees C, respectively. Heat treatment of E3d raised the fluorescence intensities of Trp residue, intrinsic FAD, and extrinsic 8-anilinonaphthalene-1-sulfonate. E3d lost FAD, and inactive E3d polypeptides were aggregated. The sulfonate bound to the aggregate became notably fluorescent. The thermal disintegration of E3d was speculated to be a consecutive reaction that was different from the concurrent disintegration reaction of the complex. Some interactions with other component polypeptides was suggested to improve the thermostability of E3c. PMID- 11055398 TI - Gene regulation in response to overexpression of cytochrome P450 and proliferation of the endoplasmic reticulum in Saccharomyces cerevisiae. AB - (CYP52A4) in Saccharomyces cerevisiae. Using the mRNA differential display technique, six genes were found to be up-regulated: ASN2, MDJ1, YLR194c, YNL208w, YER175, and YGL121c. Genes coding for Dur1.2p, Dal2p, and Sps19p were down regulated. Two strongly induced genes, which were found to accommodate the peroxisome box (YLR194c) and a 10-bp consensus sequence of genes involved in lipid metabolism (YNL208w) in their promoter regions, were further analyzed with respect to the course of induction, the necessity of the P450 membrane anchor for induction, and the effects of gene disruption on P450Cm2 overexpression. We found that both genes are not essential to overproduce P450Cm2, but their induction was dependent on P450Cm2 membrane integration. PMID- 11055399 TI - Defect in cell wall integrity of the yeast saccharomyces cerevisiae caused by a mutation of the GDP-mannose pyrophosphorylase gene VIG9. AB - The Saccharomyces cerevisiae VIG9 gene encodes GDP-mannose pyrophosphorylase, which synthesizes GDP-mannose from GTP and mannose-1-phosphate. Although the null mutant was lethal, the vig9 mutants so far obtained showed no growth defect but immature protein glycosylation and drug hypersensitivity. During our search for cell-wall mutants, we found a novel temperature-sensitive mutant, JS30, which required an osmotic stabilizer for viability. JS30 excreted cell surface proteins in the medium without any indication of cell lysis. Although conventional genetic analysis using mating was impossible, by detailed characterization of JS30 including an in vitro enzyme assay and nucleotide sequencing, we found the defect of JS30 was due to a mutation in the VIG9 gene. These results indicated a critical role of GDP-mannose in maintenance of cell-wall integrity. PMID- 11055400 TI - A positive screening for drugs that specifically inhibit the Ca2+-signaling activity on the basis of the growth promoting effect on a yeast mutant with a peculiar phenotype. AB - An inappropriate activation of a signaling pathway in yeast often has a deleterious physiological effect and causes various defects, including growth defects. In a certain genetic background (deltazds1) of Saccharomyces cerevisiae, the cell-cycle progression in G2 is specifically blocked in the medium with CaCl2 by the hyperactivation of the Ca2+-signaling pathways. Here, we developed a novel drug screening procedure designed to detect the active compounds that specifically attenuate the Ca2+-signaling activity on the basis of the ability to abrogate the growth defect of the cells suffering from the hyperactivated Ca2+ signal. Using known calcineurin inhibitors as model compounds, we have established the screening conditions for the drugs that suppress the Ca2+-induced growth inhibition. An indicator strain with an increased drug sensitivity was constructed with a syr1/erg3 null mutation. PMID- 11055401 TI - Molecular cloning and characterization of a cDNA and a gene for subtilisin-like serine proteases from rice (Oryza sativa L.) and Arabidopsis thaliana. AB - The complete nucleotide sequences of a cDNA (RSP1) that encodes a subtilisin-like serine protease (subtilase) of rice (Oryza sativa L.) and a gene (ASP48) for Arabidopsis subtilase were analyzed. The RSPI cDNA and ASP48 DNA encoded 736- and 757-residue pre-pro-polypeptides including a signal peptide with molecular masses of 78,668 Da and 79,414 Da, respectively. RSP1 is the first known serine protease in rice, and ASP48 is a gene for ara12 cDNA. Sequence comparison and phylogenetic analysis showed that RSP1 is distantly related to all other plant subtilases and ASP48 is closely related to a tomato subtilase, SBT1. The ASP48 gene was found to lack introns. The Arabidopsis subtilase gene appears to consist of a small gene family. The RSP1 was found to be expressed in seed and shoots of seedlings while ASP48 transcripts was found to be accumulated in immature silique and flowers, indicating that both RSP1 and ASP48 are organ-specific and may be involved in the specific proteolytic events that occur during organ development. PMID- 11055402 TI - Degradation of bisphenol A by the lignin-degrading enzyme, manganese peroxidase, produced by the white-rot basidiomycete, Pleurotus ostreatus. AB - Degradation of 2,2-Bis(4-hydroxyphenyl)propane (bisphenol A, BPA), an endocrine disturbing chemical, by the growing mycelia of the white-rot basidiomycete, Pleurotus ostreatus, was examined. About 80% of BPA initially present decreased in 12 days of culture with this fungus. By in vitro experiments using the lignin degrading enzyme manganese peroxidase (MnP), BPA was metabolized to phenol, 4 isopropenylphenol, 4-isopropylphenol, and hexestrol. The degradation products of BPA were assumed to be formed by the one-electron oxidation of the substrate. PMID- 11055403 TI - Enzymatic assay of histamine by amperometric detection of H2O2 with a peroxidase based sensor. AB - A method for an enzymatic assay of histamine by using histamine oxidase from Arthrobacter globiformis in combination with amperometric determination of H2O2 is described. Histamine could be quantified at a level as low as 10(-7) M. The assay is adaptable to determine histamine in food samples including tuna fish with good sensitivity and selectivity. PMID- 11055404 TI - Tyrosinase inhibitory activity of Bangladeshi indigenous medicinal plants. AB - The tyrosinase-inhibitory activity of 15 kinds of Bangladeshi medicinal plants was evaluated. Methanol extracts were prepared for screening tests, and other kinds of extracts were also studied for those with high activity. Swertia chirata, Piper nigrum, Glycyrrhiza glabra, Piper longam and Ocimum americanum were screened as highly inhibiting samples. Methanol was found to be the most efficient solvent for extracting the active compounds. The 50% tyrosinase inhibitory concentration of the Glycyrrhiza glabra methanol extract was 21.2 microg/ml. PMID- 11055405 TI - A dihydroxy-gamma-lactone as an oviposition stimulant for the swallowtail butterfly, Papilio bianor, from the rutaceous plant, Orixa japonica. AB - The oviposition response of the Rutaceae-feeding swallowtail butterfly, Papilio bianor, was induced by a methanolic extract from leaves of its major host, Orixa japonica. Several components were responsible for this oviposition response. One of the stimulants was isolated and identified as (-)-2-C-methyl-D-erythrono-1,4 lactone. The compound was inactive alone, but elicited oviposition behavior when mixed with other fractions. PMID- 11055406 TI - Practical synthesis of the disaccharide epitope, D-galactopyranosyl-alpha-1,3-D galactopyranose, by using 1,2;5,6-di-O-cyclohexylidene-alpha-D-galactofuranose as the glycosyl acceptor. AB - D-Galactosyl-alpha-1,3-D-galactopyranose (1) was chemically prepared in a good yield by coupling phenyl 2,3,4,6-tetra-O-benzyl-1-thio-beta-D-galactopyranoside (5) or 2,3,4,6-tetra-O-benzyl-alpha-D-galactopyranosyl bromide (8) with 1,2:5,6 di-O-cyclohexylidene-alpha-D-galactofuranose (3) with subsequent de-O-benzylation and de-O-cyclohexylidenation of the resulting protected alpha1,3-disaccharide. PMID- 11055407 TI - Purification and characterization of biliverdin-binding protein from larval hemolymph of the swallowtail butterfly, Papilio xuthus L. AB - The biliverdin-binding protein from the larval hemolymph of the swallowtail butterfly, Papilio xuthus L., was purified and characterized. The crude biliverdin-binding protein, obtained by ammonium sulfate fractionation, was purified in two steps, the first one by gel filtration chromatography and the second one by ion-exchange chromatography. The molecular mass of the purified protein was analyzed by SDS-polyacrylamide gel electrophoresis and estimated to be 21 kDa. The Namino terminal sequence of P. xuthus biliverdin-binding protein analyzed up to the 19th residue showed that 42% of the amino acid sequence are sequence similarity to the bilin-binding protein from Pieris brassicae. These results suggest that the P. xuthus biliverdin-binding protein belongs to the insecticyanin-type. PMID- 11055408 TI - On the sweetness of N-(trifluoroacetyl)aspartame. AB - A panel of tasters has found that the N-trifluoroacetyl derivative of aspartame is five times less sweet than the parent compound, contrary to the tenet in the literature, but consistent with sweet receptor models which require this nitrogen to exist in protonated form. PMID- 11055409 TI - Staurosporine promotion of formation of continuous monolayers of primary rat hepatocytes by improving attachment and spreading. AB - Primary rat hepatocytes form discontinuous monolayers even at their maximum density. Here, we show that staurosporine promotes attachment and spreading of hepatocytes onto culture substrates, so that hepatocytes form a close, continuous monolayer. This treatment did not attenuate major hepatic functions. Therefore, this technique is promising for making seamless cell-sheet structures, which will be applicable for cell-polarity experiments or artificial liver construction. PMID- 11055410 TI - Syntheses of stereochemically restricted lactone-type analogues of jasmonic acids. AB - 5-Oxa-7-epi-jasmonic acid and 5-oxa-jasmonic acid, which are stereochemically restricted lactone-type analogues of jasmonic acids, were synthesized via three component coupling of 2(5H)-furanone, tert-butyl acetate and 1-bromo-2-pentyne. After acidic deprotection of the tert-butyl esters, the (Z)-olefin was introduced by catalytic partial reduction with the Lindlar catalyst to give the desired analogues. PMID- 11055411 TI - Recognition of receptor lipopolysaccharides by spike G protein of bacteriophage phiX174. AB - The spike G protein of bacteriophage phiX174 was prepared as a hexa histidine tagged G protein (HisG). In the enzyme-linked plate assay, HisG bound specifically to lipopolysaccharides (LPSs) of the phiX174-sensitive strains, and did not bind to LPSs of the phiX174-insensitive strains. The truncated G protein obtained after trypsin digestion of HisG had the similar affinity to the LPSs to HisG, indicating that eight amino acid residues from the N-terminus are not essential to the binding with the LPSs. PMID- 11055412 TI - Fungicidal and herbicidal activities of berberine related alkaloids. AB - The 23 quaternary and tertiary protoberberines related to berberine were tested for in vitro and/or in vivo fungicidal and herbicidal activities. Among the compounds tested, there was some activity observed with some of only the protoberberinium salts, but not sufficiently strong or broad spectrum for agrochemical use. From the structure-activity point of view, some features can be pointed out. PMID- 11055413 TI - Suppression of D-galactosamine-induced liver injury by mushrooms in rats. AB - Six species of edible mushroom were found to suppress D-galactosamine-induced enhancement of plasma alanine and aspartate aminotransferase activities when powdered mushrooms were added to the diet (5%) and fed to rats for 2 wk. Grifola frondosa exhibited the most potent effect in a dose-dependent manner. A significant effect was observed only from the water-soluble low-molecular-weight fraction of G. frondosa. The results indicate that several mushrooms possess a protective effect against liver injury induced by D-galactosamine. PMID- 11055414 TI - Producing a low ovomucoid egg white preparation by precipitation with aqueous ethanol. AB - A novel method for producing a low ovomucoid egg white preparation is proposed. Egg white powder (0.5 g) was dissolved in a 10-fold weight of distilled water and adjusted to pH 5, and ethanol was added to the solution at a final concentration of 20% (v/v). The mixture was vigorously stirred and centrifuged. The precipitate was washed three times with 20% ethanol (6.25 ml each), with about 65% of egg white proteins occurring in the precipitate. The use of ELISA demonstrated that 70% of ovomucoid was recovered from the supernatant fraction. However, functionally important proteins such as ovalbumin, ovotransferrin, and lysozyme still remained in the precipitate. These results may be due primarily to the much higher solubility of ovomucoid in this aqueous ethanol. Food quality evaluation showed that high whippability and foam stability were retained in the low ovomucoid preparation as in its material egg white. This product would thus be applicable as a new processed food for ovomucoid-sensitive allergic patients. PMID- 11055415 TI - Application of a metal switch to aqualysin I, a subtilisin-type bacterial serine protease, to the S3 site residues, ser102 and gly131. AB - We applied 'metal switch' experiments to the S3 site residues, Ser102 and Gly131, of aqualysin I, a subtilisin-type serine protease. We showed that two histidines introduced at these positions did take part in histidine-metal-histidine bridge formation, and metal ions inhibited the protease activities. These results indicate that two histidines are near each other, and both side chains are metal accessible. This is the first report on application of the metal-switch technique to a subtilisin-related enzyme. PMID- 11055416 TI - High expression of the second lysine decarboxylase gene, ldc, in Escherichia coli WC196 due to the recognition of the stop codon (TAG), at a position which corresponds to the 33th amino acid residue of sigma38, as a serine residue by the amber suppressor, supD. AB - Escherichia coli WC196, which was obtained from the strain W3110 by nitrosoguanidine mutagenesis as an overproducer of lysine, produced approximately twenty times more cadaverine than did W3110, and had a twenty fold higher level of rpoS gene product, sigma38, than in W3110. Both WC196 and W3110 had a stop codon (TAG) in rpoS at position which corresponds to the 33th residue of sigma38 protein. In addition, WC196 but not W3110 had a mutation in the gene encoding Ser tRNA (SerU), called, supD. Analysis of the amino acid sequence of a sigma38 preparation from WC196 showed that the 33th residue of sigma38 is a serine residue. The deltarpoS deltacadA mutant of E. coli W3110 harboring the plasmid containing rpoS, in which the TAG codon was converted to a TCG codon for serine 33 residue of sigma38, expressed a significant amount of Ldc and accumulated a large amount of sigma38. However, the deltarpoS deltacadA mutant of W3110 with the plasmid containing the intact rpoS from W3110 could synthesize neither sigma38 nor Ldc significantly. PMID- 11055417 TI - The aman6 gene encoding a yeast mannan backbone degrading 1,6-alpha-D-mannanase in Bacillus circulans: cloning, sequence analysis, and expression. AB - A gene (aman6) encoding endo-1,6-alpha-D-mannanase, a yeast mannan backbone degrading enzyme from Bacillus circulans was cloned. The putative aman6 was 1,767 base pairs long and encoded a mature 1,6-alpha-D-mannanase protein of 589 amino acids and a signal peptide of 36 amino acids. The purified mature 1,6-alpha-D mannanase from the Escherichia coli transformant showed 61-kDa protein, and N terminal amino acid sequence and other general properties of the recombinant enzyme were identical to those of 1,6-alpha-D-mannanase from Bacillus circulans TN-31. PMID- 11055418 TI - Detection of biomarkers for apoptosis in rat liver after perfusion with 3-amino 1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1). AB - We previously demonstrated that 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp P-1) induced apoptosis in primary cultured rat hepatocytes. In this study, we investigated apoptotic biomarkers in rat liver after perfusion with 30 microM Trp P-1 as preliminary experiments for in vivo study. Induction of c-Myc and p53 protein and the activities of caspase-3, -6, and -8 were detected in Trp-P-1 perfused liver. In addition, Trp-P-1 modulated the DNA binding activity of the apoptosis-related transcription factors, NF-kappaB and AP-1. These results imply a possibility that Trp-P-1 would induce apoptosis in vivo. PMID- 11055419 TI - Illegal immigrations. PMID- 11055420 TI - Presenilins, Alzheimer's disease, and capacitative calcium entry. PMID- 11055421 TI - Reliability of neuronal responses. PMID- 11055422 TI - Narcolepsy: a neurodegenerative disease of the hypocretin system? PMID- 11055423 TI - Abeta-generating enzymes: recent advances in beta- and gamma-secretase research. PMID- 11055424 TI - Taking apart the gating of voltage-gated K+ channels. PMID- 11055425 TI - Correlated neuronal activity and visual cortical development. PMID- 11055426 TI - The millennium of the dendrite? PMID- 11055427 TI - Single molecular force spectroscopy of modular proteins in the nervous system. PMID- 11055428 TI - A genetically encoded ratiometric indicator for chloride: capturing chloride transients in cultured hippocampal neurons. AB - We constructed a novel optical indicator for chloride ions by fusing the chloride sensitive yellow fluorescent protein with the chloride-insensitive cyan fluorescent protein. The ratio of FRET-dependent emission of these fluorophores varied in proportion to the concentration of Cl and was used to measure intracellular chloride concentration ([Cl-]i) in cultured hippocampal neurons. [Cl-]i decreased during neuronal development, consistent with the shift from excitation to inhibition during maturation of GABAergic synapses. Focal activation of GABAA receptors caused large changes in [Cl-]i that could underlie use-dependent depression of GABA-dependent synaptic transmission. GABA-induced changes in somatic [Cl-]i spread into dendrites, suggesting that [Cl-]i can signal the location of synaptic activity. This genetically encoded indicator will permit new approaches ranging from high-throughput drug screening to direct recordings of synaptic Cl- signals in vivo. PMID- 11055429 TI - Active stabilization of electrodes for intracellular recording in awake behaving animals. AB - Intracellular recording is a powerful electrophysiology technique that has revealed much of what is known about the biophysical properties of neurons. However, neuronal properties are strongly affected by activity dependent and modulatory influences, making it essential, ultimately, to study these properties in behaving animals. Unfortunately, intracellular recording has only been widely applied in vitro, since cardiac and respiratory pulsations make intracellular recording difficult in vivo. In awake behaving animals, spontaneous movements make intracellular recording nearly impossible. Here I present a novel technique to dynamically stabilize the position of a recording electrode relative to the brain. Physiological signals that are predictive of brain motion at the recording site, such as the electrocardiogram (EKG), respiratory pressure, or cranial motion, are used to control a piezoelectric manipulator, making possible stable intracellular recordings in awake active animals. PMID- 11055430 TI - Reduced number of hypocretin neurons in human narcolepsy. AB - Murine and canine narcolepsy can be caused by mutations of the hypocretin (Hcrt) (orexin) precursor or Hcrt receptor genes. In contrast to these animal models, most human narcolepsy is not familial, is discordant in identical twins, and has not been linked to mutations of the Hcrt system. Thus, the cause of human narcolepsy remains unknown. Here we show that human narcoleptics have an 85%-95% reduction in the number of Hcrt neurons. Melanin-concentrating hormone (MCH) neurons, which are intermixed with Hcrt cells in the normal brain, are not reduced in number, indicating that cell loss is relatively specific for Hcrt neurons. The presence of gliosis in the hypocretin cell region is consistent with a degenerative process being the cause of the Hcrt cell loss in narcolepsy. PMID- 11055431 TI - Origin of the precerebellar system. AB - The precerebellar system provides the principal input to the cerebellum and is essential for coordinated motor activity. Using a FLP recombinase-based fate mapping approach, we provide direct evidence in the mouse that this ventral brainstem system derives from dorsally located rhombic neuroepithelium. Moreover, by fate mapping at the resolution of a gene expression pattern, we have uncovered an unexpected subdivision within the precerebellar primordium: embryonic expression of Wnt1 appears to identify the class of precerebellar progenitors that will later project mossy fibers from the brainstem to the cerebellum, as opposed to the class of precerebellar neurons that project climbing fibers. Differential gene expression therefore appears to demarcate two populations within the precerebellar primordium, grouping progenitors by their future type of axonal projection and synaptic partner rather than by final topographical position. PMID- 11055432 TI - Disruption of the type III adenylyl cyclase gene leads to peripheral and behavioral anosmia in transgenic mice. AB - Cyclic nucleotide-gated ion channels in olfactory sensory neurons (OSNs) are hypothesized to play a critical role in olfaction. However, it has not been demonstrated that the cAMP signaling is required for olfactory-based behavioral responses, and the contributions of specific adenylyl cyclases to olfaction have not been defined. Here, we report the presence of adenylyl cyclases 2, 3, and 4 in olfactory cilia. To evaluate the role of AC3 in olfactory responses, we disrupted the gene for AC3 in mice. Interestingly, electroolfactogram (EOG) responses stimulated by either cAMP- or inositol 1,4,5-triphosphate- (IP3-) inducing odorants were completely ablated in AC3 mutants, despite the presence of AC2 and AC4 in olfactory cilia. Furthermore, AC3 mutants failed several olfaction based behavioral tests, indicating that AC3 and cAMP signaling are critical for olfactory-dependent behavior. PMID- 11055433 TI - Spatially and functionally distinct roles of the PI3-K effector pathway during NGF signaling in sympathetic neurons. AB - NGF is a target-derived growth factor for developing sympathetic neurons. Here, we show that application of NGF exclusively to distal axons of sympathetic neurons leads to an increase in PI3-K signaling in both distal axons and cell bodies. In addition, there is a more critical dependence on PI3-K for survival of neurons supported by NGF acting exclusively on distal axons as compared to neurons supported by NGF acting directly on cell bodies. Interestingly, PI3-K signaling within both cell bodies and distal axons contributes to survival of neurons. The requirement for PI3-K signaling in distal axons for survival may be explained by the finding that inhibition of PI3-K in the distal axons attenuates retrograde signaling. Therefore, a single TrkA effector, PI3-K, has multiple roles within spatially distinct cellular locales during retrograde NGF signaling. PMID- 11055434 TI - The murine cone photoreceptor: a single cone type expresses both S and M opsins with retinal spatial patterning. AB - Mice express S and M opsins that form visual pigments for the detection of light and visual signaling in cones. Here, we show that S opsin transcription is higher than that of M opsin, which supports ultraviolet (UV) sensitivity greater than midwavelength sensitivity. Surprisingly, most cones coexpress both S and M opsins in a common cone cell type throughout the retina. All cones express M opsin, but the levels are graded from dorsal to ventral. The levels of S opsin are relatively constant. However, in the far dorsal retina, S opsin is repressed stochastically, such that some cones express M opsin only. These observations indicate that two different mechanisms control M and S opsin expression. We suggest that a common cone type is patterned across the retinal surface to produce phenotypic cone subtypes. PMID- 11055435 TI - The gain of rod phototransduction: reconciliation of biochemical and electrophysiological measurements. AB - We have resolved a central and long-standing paradox in understanding the amplification of rod phototransduction by making direct measurements of the gains of the underlying enzymatic amplifiers. We find that under optimized conditions a single photoisomerized rhodopsin activates transducin molecules and phosphodiesterase (PDE) catalytic subunits at rates of 120-150/s, much lower than indirect estimates from light-scattering experiments. Further, we measure the Michaelis constant, Km, of the rod PDE activated by transducin to be 10 microM, at least 10-fold lower than published estimates. Thus, the gain of cGMP hydrolysis (determined by kcat/Km) is at least 10-fold higher than reported in the literature. Accordingly, our results now provide a quantitative account of the overall gain of the rod cascade in terms of directly measured factors. PMID- 11055436 TI - Actin-dependent regulation of neurotransmitter release at central synapses. AB - Depolymerization of actin by latrunculin A transiently promotes neurotransmitter release. The mean rate of mEPSCs increases by a Ca2+-independent process, without a concomitant change in the mean amplitude. The readily releasable vesicle pool size and the rate of refilling of the readily releasable pool remain unaltered by latrunculin treatment. Evoked neurotransmitter release also increases in a manner consistent with an increase in vesicle release probability. The observed enhancement of neurotransmitter release is specific to actin depolymerization mediated by latrunculin A and is not caused by cytochalasin D. Our findings indicate that actin participates in a regulatory mechanism that restrains fusion of synaptic vesicles at the active zone. PMID- 11055437 TI - Two endocytic recycling routes selectively fill two vesicle pools in frog motor nerve terminals. AB - We have identified and characterized two vesicle recycling pathways in frog motor nerve terminals. We exploited the differential staining properties of FM dyes of varying hydrophobicity to label selectively two different vesicle pools, using optical imaging and electron microscopy of photoconverted dyes. During a 1 min tetanus, a rapidly recycling route places vesicles selectively into a small readily releasable pool comprising about 20% of vesicles. After the tetanus, a much slower pathway (from which FM2-10 but not FM1-43 can be rinsed) delivers vesicles via infoldings and cisternae selectively to a reserve pool with a halftime of about 8 min. Mixing between the two pools is slow. During stimulation at 30 Hz, 10-15 s is required to mobilize and release dye from the reserve pool. PMID- 11055438 TI - Presenilin-mediated modulation of capacitative calcium entry. AB - We studied a novel function of the presenilins (PS1 and PS2) in governing capacitative calcium entry (CCE), a refilling mechanism for depleted intracellular calcium stores. Abrogation of functional PS1, by either knocking out PS1 or expressing inactive PS1, markedly potentiated CCE, suggesting a role for PS1 in the modulation of CCE. In contrast, familial Alzheimer's disease (FAD) linked mutant PS1 or PS2 significantly attenuated CCE and store depletion activated currents. While inhibition of CCE selectively increased the amyloidogenic amyloid beta peptide (Abeta42), increased accumulation of the peptide had no effect on CCE. Thus, reduced CCE is most likely an early cellular event leading to increased Abeta42 generation associated with FAD mutant presenilins. Our data indicate that the CCE pathway is a novel therapeutic target for Alzheimer's disease. PMID- 11055439 TI - A conserved glutamate is important for slow inactivation in K+ channels. AB - Voltage-gated ion channels undergo slow inactivation during prolonged depolarizations. We investigated the role of a conserved glutamate at the extracellular end of segment 5 (S5) in slow inactivation by mutating it to a cysteine (E418C in Shaker). We could lock the channel in two different conformations by disulfide-linking 418C to two different cysteines, introduced in the Pore-S6 (P-S6) loop. Our results suggest that E418 is normally stabilizing the open conformation of the slow inactivation gate by forming hydrogen bonds with the P-S6 loop. Breaking these bonds allows the P-S6 loop to rotate, which closes the slow inactivation gate. Our results also suggest a mechanism of how the movement of the voltage sensor can induce slow inactivation by destabilizing these bonds. PMID- 11055440 TI - Reconstructing voltage sensor-pore interaction from a fluorescence scan of a voltage-gated K+ channel. AB - X-ray crystallography has made considerable recent progress in providing static structures of ion channels. Here we describe a complementary method-systematic fluorescence scanning-that reveals the structural dynamics of a channel. Local protein motion was measured from changes in the fluorescent intensity of a fluorophore attached at one of 37 positions in the pore domain and in the S4 voltage sensor of the Shaker K+ channel. The local rearrangements that accompany activation and slow inactivation were mapped onto the homologous structure of the KcsA channel and onto models of S4. The results place clear constraints on S4 location, voltage-dependent movement, and the mechanism of coupling of S4 motion to the operation of the slow inactivation gate in the pore domain. PMID- 11055441 TI - Control of dendritic outputs of inhibitory interneurons in the lateral geniculate nucleus. AB - The thalamic relay to neocortex is dynamically gated. The inhibitory interneuron, which we have studied in the lateral geniculate nucleus, is important to this process. In addition to axonal outputs, these cells have dendritic terminals that are both presynaptic and postsynaptic. Even with action potentials blocked, activation of ionotropic and metabotropic glutamate receptors on these terminals increases their output, whereas activation of metabotropic (M2 muscarinic) but not nicotinic cholinergic receptors decreases their output. These actions can strongly affect retinogeniculate transmission. PMID- 11055442 TI - Reversible associative depression and nonassociative potentiation at a parallel fiber synapse. AB - The electrosensory lobe (ELL) of mormyrid electric fish is one of several cerebellum-like sensory structures in fish that remove predictable features of the sensory inflow. This adaptive process obeys anti-Hebbian rules and appears to be mediated by associative depression at the synapses between parallel fibers and Purkinje-like cells of ELL. We show here that there is also a nonassociative potentiation at this synapse that depends only on the repeated occurrence of the EPSP. The depression can be reversed by the potentiation and vice versa. Finally, we show that the associative depression requires NMDA receptor activation, changes in postsynaptic calcium, and the occurrence of a postsynaptic dendritic spike within a few milliseconds following EPSP onset. PMID- 11055443 TI - Hippocampal neurons encode information about different types of memory episodes occurring in the same location. AB - Firing patterns of hippocampal complex-spike neurons were examined for the capacity to encode information important to the memory demands of a task even when the overt behavior and location of the animal are held constant. Neuronal activity was recorded as rats continuously alternated left and right turns from the central stem of a modified T maze. Two-thirds of the cells fired differentially as the rat traversed the common stem on left-turn and right-turn trials, even when potentially confounding variations in running speed, heading, and position on the stem were taken into account. Other cells fired differentially on the two trial types in combination with behavioral and spatial factors or appeared to fire similarly on both trial types. This pattern of results suggests that hippocampal representations encode some of the information necessary for representing specific memory episodes. PMID- 11055444 TI - Low response variability in simultaneously recorded retinal, thalamic, and cortical neurons. AB - The response of a cortical cell to a repeated stimulus can be highly variable from one trial to the next. Much lower variability has been reported of retinal cells. We recorded visual responses simultaneously from three successive stages of the cat visual system: retinal ganglion cells (RGCs), thalamic (LGN) relay cells, and simple cells in layer 4 of primary visual cortex. Spike count variability was lower than that of a Poisson process at all three stages but increased at each stage. Absolute and relative refractory periods largely accounted for the reliability at all three stages. Our results show that cortical responses can be more reliable than previously thought. The differences in reliability in retina, LGN, and cortex can be explained by (1) decreasing firing rates and (2) decreasing absolute and relative refractory periods. PMID- 11055445 TI - Reaches to sounds encoded in an eye-centered reference frame. AB - A recent hypothesis suggests that neurons in the lateral intraparietal area (LIP) and the parietal reach region (PRR) encode movement plans in a common eye centered reference frame. To test this hypothesis further, we examined how PRR neurons encode reach plans to auditory stimuli. We found that PRR activity was affected by eye and initial hand position. Population analyses, however, indicated that PRR neurons were affected more strongly by eye position than by initial hand position. These eye position effects were appropriate to maintain coding in eye coordinates. Indeed, a significant population of PRR neurons encoded reaches to auditory stimuli in an eye-centered reference frame. These results extend the hypothesis that, regardless of the modality of the sensory input or the eventual action, PRR and LIP neurons represent movement plans in a common, eye-centered representation. PMID- 11055446 TI - Children's health care issues: a continuing priority. PMID- 11055447 TI - How does risk sharing between employers and a managed behavioral health organization affect mental health care? AB - OBJECTIVE: To study the ways in which allocating the risk for behavioral health care expenses between employers and a managed behavioral health organization affects costs and the use of services. DATA SOURCES: Claims from 87 plans that cover mental health and substance abuse services covering over one million member years in 1996/1997. STUDY DESIGN: Multi-part regression models for health care cost are used. Dependent variables are health care costs decomposed into access to any care, costs per user, any inpatient use, costs per outpatient user, and costs per inpatient user. The study compares full-risk plans, in which the managed care organization provides managed care services and acts as the insurer by assuming the risk for claims costs, with contracts in which the managed care organization only manages care (for a fixed administrative fee) and the employer retains the risk for claims. PRINCIPAL FINDINGS: Full-risk plans are not statistically significantly different from non-risk plans in terms of any mental health specialty use or hospitalization rates, but costs per user are significantly lower, in particular for inpatients. CONCLUSIONS: Risk contracts do not affect initial access to mental health specialty care or hospitalization rates, but patients in risk contracts have lower costs, either because of lower intensity of care or because they are treated by less expensive providers. PMID- 11055448 TI - Primary care satellite clinics and improved access to general and mental health services. AB - OBJECTIVES: To evaluate the relationship between the implementation of community based primary care clinics and improved access to general health care and/or mental health care, in both the general population and among people with disabling mental illness. STUDY SETTING: The 69 new community-based primary care clinics in underserved areas, established by the Department of Veterans Affairs (VA) between the last quarter of FY 1995 and the second quarter of FY 1998, including the 21 new clinics with a specialty mental health care component. DATA SOURCES: VA inpatient and outpatient workload files, 1990 U.S. Census data, and VA Compensation and Pension files were used to determine the proportion of all veterans, and the proportion of disabled veterans, living in each U.S. county who used VA general health care services and VA mental health services before and after these clinics began operation. DESIGN: Analysis of covariance was used to compare changes, from late FY 1995 through early FY 1998, in access to VA services in counties in which new primary care clinics were located, in counties in which clinics that included specialized mental health components were located, and for comparison, in other U.S. counties, adjusting for potentially confounding factors. KEY FINDINGS: Counties in which new clinics were located showed a significant increase from the FY 1995-FY 1998 study dates in the proportion of veterans who used general VA health care services. This increase was almost twice as large as that observed in comparison counties (4.2% vs. 2.5%: F = 12.6, df = 1,3118, p = .0004). However, the introduction of these clinics was not associated with a greater use of specialty VA mental health services in the general veteran population, or of either general health care services or mental health services among veterans who received VA compensation for psychiatric disorders. In contrast, in counties with new clinics that included a mental health component the proportion of veterans who used VA mental health services increased to almost three times the proportion in comparison counties (0.87% vs. 0.31%: F = 8.3, df = 1,3091, p = .004). CONCLUSIONS: Community-based primary care clinics can improve access to general health care services, but a specialty mental health care component appears to be needed to improve access to mental health services. PMID- 11055449 TI - The outcome and cost of alcohol and drug treatment in an HMO: day hospital versus traditional outpatient regimens. AB - OBJECTIVE: To compare outcome and cost-effectiveness of the two primary addiction treatment options, day hospitals (DH) and traditional outpatient programs (OP) in a managed care organization, in a population large enough to examine patient subgroups. DATA SOURCES: Interviews with new admissions to a large HMO's chemical dependency program in Sacramento, California between April 1994 and April 1996, with follow-up interviews eight months later. Computerized utilization and cost data were collected from 1993 to 1997. STUDY DESIGN: Design was a randomized control trial of adult patients entering the HMO's alcohol and drug treatment program (N = 668). To examine the generalizability of findings as well as self selection factors, we also studied patients presenting during the same period who were unable or unwilling to be randomized (N = 405). Baseline interviews characterized type of substance use, addiction severity, psychiatric status, and motivation. Follow-up interviews were conducted at eight months following intake. Breathanalysis and urinalysis were conducted. Program costs were calculated. DATA COLLECTION: Interview data were merged with computerized utilization and cost data. PRINCIPAL FINDINGS: Among randomized subjects, both study arms showed significant improvement in all drug and alcohol measures. There were no differences overall in outcomes between DH and OP, but DH subjects with midlevel psychiatric severity had significantly better outcomes, particularly in regard to alcohol abstinence (OR = 2.4; 95% CI = 1.2, 4.9). The average treatment costs were $1,640 and $895 for DH and OP programs, respectively. In the midlevel psychiatric severity group, the cost of obtaining an additional person abstinent from alcohol in the DH cohort was approximately $5,464. Among the 405 self selected subjects, DH was related to abstinence (OR = 2.1; 95% CI = 1.3, 3.5). CONCLUSIONS: Although significant benefits of the DH program were not found in the randomized study, DH treatment was associated with better outcomes in the self-selected group. However, for subjects with mid-level psychiatric severity in both the randomized and self-selected samples, the DH program produced higher rates of abstention and was more cost-effective. Self-selection in studies that randomize patients to services requiring very different levels of commitment may be important in interpreting findings for clinical practice. PMID- 11055450 TI - Does the disbursement of income increase psychiatric emergencies involving drugs and alcohol? AB - OBJECTIVE: To determine if the incidence of psychiatric emergencies involving drugs or alcohol supports the argument that mentally ill persons contribute to elevated mortality during the days following disbursement of private earnings and public income transfers. STUDY DESIGN: Interrupted time-series using Box-Jenkins methods. DATA COLLECTION/EXTRACTION METHODS: Daily counts of adults admitted to psychiatric emergency services in San Francisco after using drugs or alcohol were derived from medical records for the period January 1 through June 30, 1997. PRINCIPAL FINDINGS: Psychiatric emergencies among males who had used drugs or alcohol were elevated in the early days of the month. Such emergencies among females were not similarly elevated. Emergencies among females who had not used drugs or alcohol were elevated in the early days of the month. CONCLUSION: Elevated mortality in the first week of the month may be attributable, in part, to the "check effect" or use of drugs and alcohol by mentally ill males in the days after they receive income. The contribution of women is more complex and may be induced by drug or alcohol abuse among persons in their social networks. The check effect suggests that persons with a history of substance abuse and mental illness should be offered the opportunity to have their income managed by someone who can monitor and influence how the money is being spent. The fact that drug- or alcohol-related admissions among males exhibit temporal patterns suggests that the provision of preventive as well as treatment services may be strategically scheduled. PMID- 11055451 TI - HMO growth and the geographical redistribution of generalist and specialist physicians, 1987-1997. AB - OBJECTIVE: To assess the impact of the growth in HMO penetration in different metropolitan areas on the change in the number of generalists, specialists, and total physicians, and on the change in the proportion of physicians who are generalists. DATA SOURCES/STUDY SETTING: The American Medical Association Physician Masterfile, to obtain the number of patient care generalists and specialists in 1987 and in 1997 who were practicing in each of 316 metropolitan areas in the United States. Additional data for each metropolitan area were obtained from a variety of sources, and included HMO penetration in 1986 and 1996. STUDY DESIGN: We estimated multivariate regression models in which the change in the number of physicians between 1987 and 1997 was a function of HMO penetration in 1986, the change in HMO penetration between 1986 and 1996, population characteristics and physician fees in 1986, and the change in population characteristics and fees between 1986 and 1996. Each model was estimated using ordinary least squares (OLS) and two-stage least squares (TSLS). PRINCIPAL FINDINGS: HMO penetration did not affect the number of generalist physicians or hospital-based specialists, but faster HMO growth led to smaller increases in the numbers of medical/surgical specialists and total physicians. Faster HMO growth also led to larger increases in the proportion of physicians who were generalists. Our best estimate is that an increase in HMO penetration of .10 between 1986 and 1996 reduced the rate of increase in medical/surgical specialists by 10.3 percent and reduced the rate of increase in total physicians by 7.2 percent. CONCLUSIONS: The findings of this study support the notion that HMOs reduce the demand for physician services, particularly for specialists' services. The findings also imply that, during the past decade, there has been a redistribution of physicians-especially medical/surgical specialists-from metropolitan areas with high HMO penetration to low-penetration areas. PMID- 11055452 TI - The effect of selective contracting on hospital costs and revenues. AB - OBJECTIVE: To examine the effects of selective contracting on California hospital costs and revenues over the 1983-1997 period. DATA SOURCES/STUDY SETTING: Annual disclosure data and discharge data sets for 421 California general acute care hospitals from 1980 to 1997. ANALYSIS: Using measures of competition developed from patient-level discharge data, and financial and utilization measures from the disclosure data, we estimated a fixed effect multivariate regression model of hospital costs and revenues. FINDINGS: We found that hospitals in more competitive areas had a substantially lower rate of increase in both costs and revenues over this extended period of time. For-profit hospitals lowered their costs and revenues after selective contracting was initiated relative to the cost and revenue levels of not-for-profit hospitals. The Medicare PPS has also led high-cost hospitals to lower their costs. CONCLUSIONS: The more competitive the hospital's market, the greater degree to which it has had to lower the rate of increase in costs. A similar pattern exists with regard to hospital revenues. Both of these trends appear to result from the growth of selective contracting. It remains unclear to what extent these cost reductions were the result of increased efficiency or of reduced quality. Since hospital cost growth is sensitive to the competitiveness of its market, antitrust enforcement is a critical element in any cost containment policy. PMID- 11055453 TI - The validity of information on "race" and "Hispanic ethnicity" in California birth certificate data. AB - OBJECTIVE: To evaluate the validity of racial/ethnic information in California birth certificate data. DATA SOURCES: Computerized birth certificate data and postpartum interviews with California mothers. STUDY DESIGN AND DATA COLLECTION: Birth certificates were matched with face-to-face structured postpartum interviews with 7,428 mothers to compare racial/ethnic information between the two data sources. Interviews were conducted in Spanish or English during delivery stays at 16 California hospitals, 1994-1995. PRINCIPAL FINDINGS: The sensitivity of racial/ethnic classification in birth certificate data was very high (94 percent to 99 percent) for African Americans, Asians/Pacific Islanders, Europeans/Middle Easterners, and Latinas (Hispanics). For Native Americans, however, the sensitivity was only 54 percent. The positive predictive value of birth certificate classification of race/ethnicity was high for all racial/ethnic groups (96 percent to 97 percent). CONCLUSIONS: Despite limited training of birth clerks, the maternal racial/ethnic information in California birth certificate data appears to be a valid measure of self-identified race and Hispanic ethnicity for groups other than Native Americans. PMID- 11055454 TI - The SF-12 as a population health measure: an exploratory examination of potential for application. AB - OBJECTIVE: To describe the relationships among functional health status measures (SF-12 physical and mental components summary scores), traditional measures of community health status, and social determinants of health among respondents to community health status surveys conducted in nine different communities. DATA SOURCES/STUDY SETTING: Data collected as part of comprehensive community health status assessments conducted in each of nine communities (in seven states) between 1992 and 1997. The purpose of each assessment was to gather data to plan and evaluate population health improvement initiatives. STUDY DESIGN AND DATA COLLECTION: This is an opportunistic study drawing on the universe of community health survey data collected by the authors to support local health improvement initiatives. Both community-level as well as an aggregate of individual-level measures are used in the analysis. Within each locality, survey respondents were randomly selected using a telephone-facilitated, mailed survey methodology. PRINCIPAL FINDINGS: The key variables reported here are functional health status measures (SF-12) and social determinants of health variables. SF-12 physical and mental component scales correlated with two of four traditional measures of community health status. At the aggregate level of analysis, significant relationships were found for seven of nine social determinants of health measures when compared with SF-12 component summary scores. Relationships between social determinants measures and PCS-12 and MCS-12 scores suggest both application possibilities and the need for additional analysis in order to understand the nature of those relationships. CONCLUSIONS: Physical and mental health functioning summary scores as measured by the SF-12 are useful in describing overall community health status when compared with traditional measures such as total deaths, age-adjusted mortality, or physician to population ratio. The SF-12 can also be used to measure the relationship between physical and mental health functioning (as proxies for community health status) and the social determinants of health. This analysis can help to refine our understanding of how social determinants and health status interact in a community or population as a precursor to the development of models of community or population health. PMID- 11055455 TI - Enhancing social skills of kindergarten children with autism through the training of multiple peers as tutors. AB - Many students with autism are being served in inclusive settings. Early intervention programs, traditionally home-based, are beginning to create center based options which incorporate typically developing peers. One of the arguments for the use of inclusive programs is that students with autism will benefit from their exposure to and interactions with typical peers. Unfortunately, research suggests that in inclusive settings, typical peers and peers with autism do not always interact without prompting from an adult. This study used an ABAB design to determine if a peer buddy approach in which all students were trained to interact in dyads would increase non-adult-directed interactions. Data collected on the students with autism indicate that the peer buddy approach significantly increased their appropriate social interactions. Follow-up data on one of the students indicates generalization of appropriate social interactions to a new classroom. PMID- 11055456 TI - Understanding atypical emotions among children with autism. AB - Children with autism are said to be poor mind readers: They have a limited understanding of the role that mental states play in determining emotions and behavior. In this research, 23 high-functioning children from the autistic spectrum (M age 9 years 3 months), 42 6-year-old controls, and 43 10-year-old controls were presented with six emotion-evoking stories and they were asked to explain protagonists' typical and atypical emotions. In the case of typical emotions, as expected on the basis of the mindblind hypothesis, children from the autistic spectrum gave few mental state explanations, referring to fewer than even the 6-year-old control group. However, in the case of atypical emotions, the autistic group performed as well as the 10-year-old controls. Their explanations for the atypical emotions demonstrate that children from the autistic spectrum indeed have the capacity to mind read (with respect to both desires and beliefs), although they do not always use this capacity in the same way as normally developing children. It is argued that the mind-reading capacity of high functioning children from the autistic spectrum might be basically intact; unused in everyday circumstances but not necessarily defective. PMID- 11055457 TI - The autism diagnostic observation schedule-generic: a standard measure of social and communication deficits associated with the spectrum of autism. AB - The Autism Diagnostic Observation Schedule-Generic (ADOS-G) is a semistructured, standardized assessment of social interaction, communication, play, and imaginative use of materials for individuals suspected of having autism spectrum disorders. The observational schedule consists of four 30-minute modules, each designed to be administered to different individuals according to their level of expressive language. Psychometric data are presented for 223 children and adults with Autistic Disorder (autism), Pervasive Developmental Disorder Not Otherwise Specified (PDDNOS) or nonspectrum diagnoses. Within each module, diagnostic groups were equivalent on expressive language level. Results indicate substantial interrater and test-retest reliability for individual items, excellent interrater reliability within domains and excellent internal consistency. Comparisons of means indicated consistent differentiation of autism and PDDNOS from nonspectrum individuals, with some, but less consistent, differentiation of autism from PDDNOS. A priori operationalization of DSM-IV/ICD-10 criteria, factor analyses, and ROC curves were used to generate diagnostic algorithms with thresholds set for autism and broader autism spectrum/PDD. Algorithm sensitivities and specificities for autism and PDDNOS relative to nonspectrum disorders were excellent, with moderate differentiation of autism from PDDNOS. PMID- 11055458 TI - The Awkward Moments Test: a naturalistic measure of social understanding in autism. AB - Details are given of a new advanced theory of mind task, developed to approximate the demands of real-life mentalizing in able individuals with autism. Excerpts of films showing characters in social situations were presented, with participants required to answer questions on characters' mental states and on control, nonsocial questions. When compared with control participants, adults with high functioning autism and Asperger syndrome were most impaired in their ability to answer the questions requiring mind-reading ability. Although the present findings have implications for task modification, such naturalistic, dynamic stimuli are held to offer an important means of studying subtle difficulties in mentalistic understanding. PMID- 11055459 TI - Varieties of repetitive behavior in autism: comparisons to mental retardation. AB - Systematic study of abnormal repetitive behaviors in autism has been lacking despite the diagnostic significance of such behavior. The occurrence of specific topographies of repetitive behaviors as well as their severity was assessed in individuals with mental retardation with and without autism. The occurrence of each behavior category, except dyskinesias, was higher in the autism group and autistic subjects exhibited a significantly greater number of topographies of stereotypy and compulsions. Both groups had significant patterns of repetitive behavior co-occurrence. Autistic subjects had significantly greater severity ratings for compulsions, stereotypy, and self-injury. Repetitive behavior severity also predicted severity of autism. Although abnormal repetition is not specific to autism, an elevated pattern of occurrence and severity appears to characterize the disorder. PMID- 11055460 TI - Efficacy of methylphenidate among children with autism and symptoms of attention deficit hyperactivity disorder. AB - Thirteen children (ages 5.6 to 11.2 years) with autism and symptoms of attention deficit hyperactivity disorder (ADHD) participated in a double-blind, placebo controlled crossover study of methylphenidate (0.3 and 0.6 mg/kg per dose). Eight subjects responded positively, based upon a minimum 50% decrease on the Conners Hyperactivity Index. Ratings of stereotypy and inappropriate speech, which are often associated with autistic core features, also decreased. However, no changes were found on the Child Autism Rating Scale, a global assessment of autistic symptomotology. Significant adverse side effects occurred in some children including social withdrawal and irritability, especially at the 0.6 mg/kg dose. Results suggest that methylphenidate can be efficacious for children with autism and ADHD symptoms. However, this group of children seems to be particularly susceptible to adverse side effects. PMID- 11055461 TI - Birth patterns in mentally retarded autistic patients. AB - Some studies claim to have shown that, compared to the general population, autistic children are born more often in the spring. The current study sought to replicate this finding in a large Dutch sample of mentally retarded autistic patients. Birth data for 1,031 patients with a diagnosis of "Infantile Autism" or "other psychoses with origin specific to childhood" were compared to those of the Dutch national population. Separate analyses were performed on diagnostic subgroups (i.e., infantile autism vs. other psychoses with origin specific to childhood), gender, and intelligence. No evidence was found to suggest that autism is characterized by a deviant birth pattern. PMID- 11055462 TI - Attention-deficit hyperactivity disorder (ADHD), Tourette disorder (TD), and autism. PMID- 11055463 TI - 2-year-old boy who has been diagnosed with autism. PMID- 11055464 TI - The problem of hypertension in the elderly. AB - Since most developed countries have an ageing population, the prevalence of hypertension is increasing. This age-driven increase in cardiovascular risk is an important factor contributing to the increasing burden of mortality and morbidity associated with cardiovascular disease. Today, there is a strong rationale for an aggressive approach to hypertension since antihypertensive treatment has been shown to reduce cardiovascular mortality and morbidity in the elderly. It is likely that increasing emphasis will be placed on control of isolated and borderline systolic hypertension, which are the predominant forms of hypertension in elderly patients. The recent second Swedish Trial in Old Patients with Hypertension (STOP-Hypertension-2) represents an important contribution to the literature since it shows that newer antihypertensive agents, such as angiotensin converting enzyme (ACE) inhibitors and calcium antagonists, are as effective as older agents in reducing cardiovascular mortality and morbidity in elderly patients. PMID- 11055465 TI - Target blood pressure in elderly hypertensive patients: how low should you go? AB - Epidemiological studies have highlighted the increasing prevalence of hypertension with age, and the associated increase in the risk of cardiovascular disease. A number of randomized controlled trials have shown that antihypertensive treatment significantly reduces cardiovascular mortality and morbidity in elderly patients, and there is evidence that the benefit achieved is related to the extent to which blood pressure is lowered. Furthermore, a recent analysis of data from the Hypertension Optimal Treatment (HOT) Study shows that intensive therapy produces significantly greater reductions in blood pressure in elderly patients than in younger patients, without increasing the risk of adverse events. As a result, the latest management guidelines recommend that the goal of antihypertensive therapy in elderly patients should be to achieve at least high normal blood pressures (below 140/90 mmHg). Angiotensin II type 1 receptor antagonists offer a new option for antihypertensive therapy in elderly patients, and trials such as the Study on Cognition and Prognosis in the Elderly (SCOPE) are currently investigating the effect of these agents on cardiovascular mortality and morbidity in elderly hypertensive patients. PMID- 11055466 TI - The role of ambulatory blood pressure monitoring in elderly hypertensive patients. AB - There is strong evidence that ambulatory blood pressure measurements show only limited agreement with blood pressures measured in the clinic ("office" blood pressures), and are more relevant to the prognosis of hypertension. Several markers of end-organ damage, for example, have been shown to correlate more strongly with 24-h blood pressure than with office blood pressure. In addition, end-organ damage has been shown to be correlated with 24-h blood pressure variability. Ambulatory blood pressure monitoring (ABPM) has revealed a number of differences between the blood pressure profiles of elderly and younger patients. Since 24-h blood pressure control is now widely accepted as an important goal of antihypertensive therapy, ABPM has a potentially useful role in monitoring treatment in clinical trials in elderly patients. PMID- 11055467 TI - Results of the STOP-Hypertension-2 trial. AB - The second Swedish Trial in Old patients with Hypertension (STOP-Hypertension-2) was conducted to compare the effects of "newer" antihypertensive therapies (angiotensin converting enzyme [ACE] inhibitors and calcium antagonists) and established therapies (beta-blockers and diuretics) on cardiovascular mortality and morbidity in elderly hypertensive patients. A total of 6614 patients were randomized to receive conventional treatment, ACE inhibitors or calcium antagonists, and followed for a mean of 5 years. The primary endpoint was a combination of fatal stroke, fatal myocardial infarction and other fatal cardiovascular disease; secondary endpoints were a combination of fatal or non fatal stroke or myocardial infarction, and other cardiovascular mortality. The three treatments produced similar reductions in supine systolic blood pressure. There were no significant differences in the risk of cardiovascular events between patients receiving conventional therapy and those receiving newer therapies. All three treatments were well tolerated. The STOP-Hypertension-2 results thus add to the extensive literature showing the benefits of blood pressure reduction in elderly hypertensive patients. Moreover, they are consistent with current management guidelines which emphasise the importance of the achieved blood pressure reduction in the prevention of cardiovascular events. PMID- 11055468 TI - The outcome of STOP-Hypertension-2 in relation to the 1999 WHO/ISH hypertension guidelines. AB - The 1999 hypertension management guidelines issued by the World Health Organization and the International Society of Hypertension emphasize the importance of blood pressure reduction in the prevention of cardiovascular events. Furthermore, they conclude that the benefits of treatment are due to blood pressure lowering per se, rather than to any specific antihypertensive therapy. The results of the second Swedish Trial in Old Patients with Hypertension (STOP-Hypertension-2) are consistent with these recommendations, since in this trial angiotensin converting enzyme (ACE) inhibitors and calcium antagonists reduced blood pressure to the same extent as conventional therapy with beta-blockers and diuretics in elderly hypertensive patients, and the three treatments produced similar reductions in the risk of cardiovascular events. Furthermore, a first subgroup analysis of cardiovascular mortality showed that the three treatments seemed equally effective in diabetic patients. The STOP Hypertension-2 data, therefore, are fully consistent with the 1999 hypertension management guidelines, and underline the advantages offered by both older and newer antihypertensive therapies. PMID- 11055469 TI - Blood pressure measurement-bringing it all back home. PMID- 11055470 TI - Man's two environments and disorders of civilization: aspects on prevention. PMID- 11055471 TI - Interchangeability of ambulatory and office blood pressure: limitations of reproducibility and agreement. AB - A possible equivalence of office (Off) patient-recorded blood pressure (BP) and ambulatory (Amb) BP was evaluated. The criteria were the between-visits reproducibility (R) of oscillometrically measured supine office (Off), seated Off, and 24-h Amb BP and the agreement between Off and Amb data. Randomly ordered sessions were completed within 4 months in 59 untreated patients with Amb 24-h BP 136/87 (SD 14/10) mmHg and HR 72 (SD 9). R improves as SD of differences between sessions decreases with the number (n) of values included in the average so that SDD = a n(-b), where a and b are constants for each method, data sampling strategy, and group. R of a few steady Off data is better than R of a few Amb data. As n of the averaged values increases, R of Off and Amb methods converges and with n approximately 24 becomes identical. Only further increase in n of Amb data makes R of the Amb method superior than the Off method. The variably elevated initial Off BP distorts R and agreement. After approximately 6 readings, Off BP stabilizes at a lower "steady" level. "Steady" data averaged over visits are close to the Amb 24-h average from two sessions. Supine Off "steady" level is close to supine levels before sleep. The Off versus Amb method agreement improves by averaging "steady" BP from one up to four Off sessions, up to 7 "steady" Off readings over a session, and BP in both arms. At best the SBP agreement is within +/- 7.6 mmHg in 90% of cases with 24-h Amb S/D BP ranging 115-155/75-105 mmHg. The casually elevated BP can be evaluated only by the demonstrated between-visits reproducible 24-h Amb average or by the nearly equivalent average of multiple "steady" data from two to four Off visits. PMID- 11055472 TI - Comparison of self-reported home blood pressure measurements with automatically stored values and ambulatory blood pressure. AB - AIMS: To evaluate accuracy of patient-reported home blood pressure measurements (HBPM) when compared to real HBPM and their agreement with 12-h daytime ambulatory blood pressure monitoring (ABPM). MAJOR FINDINGS: Self-reported HBPM were compared to stored values of a fully automated, oscillometric blood pressure monitor with integrated memory device and 12-h daytime ambulatory monitoring in 54 patients. In most patients (n = 46, 85%) mean reported systolic or diastolic versus real HBPM differed by no more than 4 mmHg. In eight mostly uneducated patients (15%, 95% confidence interval, CI, 7-27%) means of reported and real HBPM differed by more than 4 mmHg (range 5-28 mmHg for systolic and 0-11 mmHg for diastolic blood pressure). Systolic agreement between self-reported HBPM and 12-h daytime ABPM was better for patients reporting > or = 80% than for patients reporting < 80% of measurements correctly (mean systolic difference 0 +/- 19 versus 5 +/- 14 mmHg, respectively), whereas the opposite was true concerning agreement of diastolic blood pressure values (mean diastolic difference -6 +/- 10 and -1 +/- 9 mmHg, respectively). CONCLUSION: Inadequate conclusions due to poor reporting accuracy of HBPM are possible, especially in less educated patients. Thus, ABPM or automatically stored HBPM may be preferable to self-reported HBPM in these patients. PMID- 11055473 TI - Normal responses of atrial natriuretic factor and renal tubular function to sodium loading in hypertension-prone humans. AB - BACKGROUND: In order to explore the hypothesis of an atrial natriuretic factor (ANF) deficiency in prehypertension, we compared the response to sodium loading on ANF and renal function in subjects with positive and negative histories of hypertension. METHODS: Twenty-two offspring of hypertensive parents (OH) and 20 offspring of normotensive parents (ON) were studied after 4 days of low (50 mmol/day) or high (300 mmol/day) dietary sodium intake. The diets were allocated randomly. Blood pressure (BP), renal function, plasma concentration of ANF, cyclic guanosine monophosphate (cGMP), renin, angiotensin I and II, aldosterone, endothelin and catecholamines were determined during a clearance period of 90 min on both diets. Neurohormones were measured by radioimmunoassays. Renal function was determined by simultaneous measurements of 51Cr-ethylenediaminetetraacetate (a marker of glomerular filtration rate), lithium and sodium clearances. RESULTS: Supine systolic and diastolic BPs were significantly elevated in OH, with both low and high dietary sodium intake. There was no difference in ANF and cGMP concentrations on the low sodium diet. Increasing sodium intake caused a similar increase in ANF in OH and ON but cGMP did not change significantly. As expected the activity of the renin-angiotensin-aldosterone system was decreased by enhancing sodium intake but with both low and high sodium intake plasma renin concentration was significantly higher in OH than in ON. Activation of the sympathetic nervous system with low sodium intake was indicated by a moderate increase in plasma concentrations of epinephrine and norepinephrine in both groups. The renal effects were characterized by significant increases in GFR, lithium and sodium clearances with increasing sodium intake. There were no differences between OH and ON. Estimated values of fractional proximal and distal tubular sodium reabsorption decreased significantly and in a similar way in both OH and ON. CONCLUSION: These results indicate that the renal and neuroendocrine responses to dietary sodium loading are similar in both OH and ON. The only difference was a higher BP and an elevated plasma renin concentration on both dietary regimens in OH compared with ON. In particular, in OH and ON an identical increase in plasma ANF concentration in response to sodium loading was found. Thus, this study cannot support the hypothesis of a dysregulation of ANF in hypertension-prone humans. PMID- 11055474 TI - Efficacy and tolerability of a combination tablet of candesartan cilexetil and hydrochlorothiazide in insufficiently controlled primary hypertension--comparison with a combination of losartan and hydrochlorothiazide. AB - This randomized, double-blind study compared the antihypertensive effect, safety and tolerability of a candesartan cilexetil/hydrochlorothiazide (candesartan/HCT; 16/12.5 mg) combination tablet with that of a losartan/HCT (50/12.5 mg) combination tablet in patients with mild-to-moderate primary hypertension insufficiently controlled on previous monotherapy. Men and women, aged 20-80 years, with a sitting diastolic blood pressure (DBP) > or = 90 and < or = 110 mmHg and sitting systolic blood pressure (SBP) < or = 200 mmHg during treatment with any kind of antihypertensive monotherapy for at least 4 weeks were randomized to candesartan/HCT or losartan/HCT once daily for 12 weeks. All BP measurements were performed 24 h after previous dose. Mean values and standard deviations (SD) or confidence intervals (CI) are given. A total of 340 patients were enrolled, of whom 299 (144 women and 155 men, mean age 59.5 [10.5] years) were randomized to candesartan/HCT (n = 151) or losartan/HCT (n = 148). BPs at randomization were 159.5 (15.4)/98.4 (5.8)mmHg and 160.5 (16.1)/98.5 (5.4)mmHg, respectively. There was a greater reduction in BP with candesartan/HCT than with losartan/HCT: DBP -10.4 (-11.8; -8.9) vs -7.8 (-9.3; -6.3) mmHg, difference between treatments -2.6 (-4.7; -0.5) mmHg (p = 0.016); SBP -19.4 (-22.1; -16.7) vs - 13.7 (-16.5; - 10.9) mmHg, difference between treatments -5.7 (-9.6; -1.8) mmHg (p = 0.004). The proportion of patients achieving a DBP < or = 90 mmHg was greater in the candesartan/HCT group: 60.9 (53.1; 68.7) vs 49.3 (41.3; 57.4)% (p = 0.044). There were 12 withdrawals in the candesartan/HCT group, of which 8 were due to adverse events, and 17 and 12, respectively in the losartan/HCT group. We conclude that the combination of candesartan and HCT reduces BP effectively and is well tolerated. BP was normalized in 61% of these patients who had insufficient response to previous monotherapy. The reduction in BP and the proportion of patients with normalized BP were greater with the candesartan/HCT 16/12.5 mg combination than with the losartan/ HCT 50/12.5 mg combination. PMID- 11055475 TI - Underestimation of 24-hour hypotensive efficacy of nifedipine GITS versus enalapril: ambulatory recording as an adjunct to clinical blood pressure measurement. AB - AIMS: Short-acting calcium entry blockers should be used primarily in slow release form. Furthermore, studies of the antihypertensive efficacy of drugs can be negatively influenced by between 15% and 30% of the enrolled patients not being hypertensive according to ambulatory blood pressure (BP) measurement. Thus, a randomized double-blind multicenter parallel-group study was conducted to compare the effect of nifedipine GITS (gastrointestinal therapeutic system) with enalapril. METHODS AND RESULTS: After a 2-week placebo run-in period, 186 patients with a sitting diastolic BP > or = 95 mmHg were enrolled for an 8-week treatment period. They received 30-60 mg nifedipine GITS or 5-10 mg enalapril. Diastolic BP fell comparably from 99 to 87 mmHg (p < 0.01) in the nifedipine GITS group, and from 100 to 88 mmHg (p < 0.01) in the enalapril group. The increase in BP 2 h before waking, however, was suppressed significantly more by nifedipine. Furthermore, this study highlighted the existence of "whitecoat" hypertension in a number of patients, especially when clinical BP was used to identify hypertension. Of the patients who had been identified as hypertensive before randomization by standardized BP measurement, 53 (28.5%) were identified as non hypertensives by 24-h BP monitoring. This led to an underestimation of the efficacy of the antihypertensive therapy. CONCLUSION: Nifedipine GITS as well as enalapril are comparably effective antihypertensive drugs. PMID- 11055476 TI - Testosterone increases blood pressure and cardiovascular and renal pathology in spontaneously hypertensive rats. AB - The objective of this paper was to test the hypothesis that testosterone (T) raises blood pressure (BP), which is associated with increased coronary adventitial collagen, whereas the hemodynamic force of BP increases the coronary media:lumen ratio. Five treatment groups of spontaneously hypertensive rat (SHR) were established (n = 8-10 per group): controls; hydralazine (HYZ); castration; castration + HYZ; and castration + HYZ + T + captopril. At 12 weeks of age, the castrate + HYZ group was divided so that the mean BP was the same in both groups (162 mmHg). Both groups continued to receive HYZ treatment; however one group received T implants. Also, at 12 weeks of age the castrate + HYZ + T + captopril group received T implants. BP in the HYZ group was reduced compared with controls (192 mmHg vs 218 mmHg, p < 0.01). Castration lowered BP to 170 mmHg (p < 0.01) compared with controls. However, T implants increased BP by 15 mmHg (p < 0.02) in the castrate + HYZ group and by 44 mmHg in the castrate + HYZ + captopril group (p < 0.01). Captopril in combination with HYZ significantly reduced BP compared with controls but T replacement increased BP and coronary collagen deposition in spite of HYZ and captopril treatment. PMID- 11055477 TI - The Gilda Radner Familial Ovarian Cancer Registry 1981-2000. PMID- 11055478 TI - What to do and not to do in gynecologic oncology surgery. PMID- 11055479 TI - On the evolution of a successful treatment program for a solid tumor system. PMID- 11055480 TI - Estrogen replacement therapy and ovarian cancer. AB - Estrogen replacement therapy (ERT) has been shown to be of benefit for menopausal women, especially in prevention of coronary heart disease and osteoporotic fractures. Cancer fear is an important obstacle to use of ERT. From our literature review, there is a weak or no association between ERT and ovarian cancer risk. Individual risk of cancer should be considered before ERT use. The second issue in this review is ERT in patients with ovarian cancer. The majority of patients with ovarian cancer are postmenopausal or become menopausal after surgery. ERT is considered by many physicians to be contraindicated in patients with cancer. However, there is evidence that ERT in selected cancer patients may be of benefit for survival and quality of life. After weighing the evidence from studies on ERT in patients with ovarian, breast or endometrial cancer, we propose the use of ERT in selected ovarian cancer patients who are suffering from or are at a high risk of debilitating menopausal symptoms, osteoporosis, and coronary heart disease. The benefit of ERT to selected patient's health and quality of life appears to outweigh the risk of cancer recurrence. PMID- 11055481 TI - Risk factors in stage III epithelial ovarian cancer: previous sterilisation is an adverse independent prognostic indicator. AB - OBJECTIVES: To determine whether past history of pelvic surgery is of prognostic significance in stage III epithelial ovarian cancer. METHODS: A retrospective review of 140 women with stage III epithelial ovarian cancer. RESULTS: Sixteen women had previously undergone pelvic surgery including eight sterilisations (6%), seven hysterectomies (5%) and one ovarian cystectomy (0.7%). Women with a past history of sterilisation were significantly younger (median age, 46 years) than women without a past history of sterilisation (median age, 63 years), and also significantly younger than women with a past history of hysterectomy (median age, 58 years). In addition, the sterilisation procedure was performed at a significantly younger age than the hysterectomy procedure (p=0.008). On multivariate analysis comparing previous pelvic surgery, previous malignancy, place of surgery, interval/secondary debulking, presence of concomitant tumour, performance of bowel surgery, histological grade, histological type, size of residual disease and age, all of the following were seen to be independent variables associated with outcome survival; previous sterilisation (p=0.0012), age (p=0.0074), histological type (p=0.025), histological grade (p=0.0017) and size of residual disease (p=0.0043). CONCLUSION: Past history of sterilisation appears to be an adverse independent prognostic indicator in women presenting with stage III epithelial ovarian cancer. To have developed ovarian cancer despite the protective effects of a sterilisation procedure against environmental factors might possibly suggest a predisposition to ovarian cancer in these women. Further studies are indicated to confirm the present results. PMID- 11055482 TI - Meigs syndrome and "false positive" preoperative serum CA-125 levels: analysis of ten cases. AB - Ten patients with Meigs syndrome operated on by the author were evaluated to determine the possible relationship between elevated preoperative serum CA-125 levels and tumor size and/or ascites. PMID- 11055483 TI - Primary fallopian tube carcinoma with isolated torsion of involved tube. AB - BACKGROUND: Primary fallopian tube carcinoma (FTC) is an aggressive but rare tumor. Worldwide, more than 1,500 cases have been published, and about 20 new cases are added every year. Isolated fallopian tube torsion (IFTT) is an unusual and uncommon event. CASE: We report a 69-year-old Caucasian woman, Gravida 4, Para 3, with a long history of hypertension, diabetes mellitus with retinopathy and neuropathy, and history of extensive coronary artery disease, for which a triple-by-pass graft was performed. She was placed on anticoagulation therapy. Subsequently, she developed intermittent vaginal bleeding. RESULTS: We reviewed and discussed the symptoms and work-up of the patient in detail. She underwent exploratory laparotomy, and primary FTC with isolated torsion of the involved fallopian tube was diagnosed. Peritoneal washings, omentectomy, total abdominal hysterectomy and bilateral salpingo-oophorectomy were performed. CONCLUSION: Review of the English literature on the presenting symptoms and diagnostic management of primary FTC and IFTT is presented. PMID- 11055484 TI - Colposcopic scoring system for biopsy decisions in different patient groups. AB - Colposcopic scoring system have been used for distinguishing low-grade from high grade cervical lesions. However, none of the previous studies have reported colposcopic scoring systems for biopsy decisions in different patient groups. The purpose of our study was to evaluate the safety of biopsy decisions using the colposcopic score elaborated by Stellato and Paavonen (IL) in 21 nonpregnant HIV infected patients (NP+) and 36 uninfected patients (NP-), 12 HIV infected pregnant patients (P+) and 20 uninfected pregnant patients (P-) in the diagnosis of cervical intraepithelial neoplasias (CIN) and HPV infection. The receiving operator curve was used for the establishment of a cut-off point in the scoring system graduation. The chi-square test was used for the statistical analysis. We obtained a safety cut-off value in the colposcopic scoring system for each patient group: 4.5 for NP+; 4.0 for NP- and 3.5 for P+ and P- patients. The sensitivity and specificity of the colposcopic score for the detection of high grade lesions for each group were respectively: 87.5 and 92.3% for NP+ patients; 90.9 and 92% for NP- patients; 100 and 87.5% for P+ patients and 100 and 91.7% for P- patients. Our results suggest that the colposcopic scoring system is a practical tool for a colposcopy-guided punch biopsy decision and detection of high-grade cervical lesions in different patient groups. Further studies are needed to prove its clinical utility. PMID- 11055485 TI - Rhabdomyosarcoma of the corpus of the uterus: a case report. AB - A case of uterine rhabdomyosarcoma is presented. Preoperative chemotherapy resulted in an excellent response. PMID- 11055486 TI - Micropapillary serous carcinoma of the ovary: case report and review of literature. AB - The term "micropapillary serous carcinoma" (MPSC) has recently been introduced to define a subset of ovarian serous borderline tumors morphologically characterized by a micropapillary pattern and clinically associated with a more aggressive behavior than that of the typical ovarian serous borderline tumors. Ovarian MPSC's are associated with extra-ovarian invasive peritoneal implants and invasive recurrences much more frequently than typical ovarian serous borderline tumors. The case of a women, who at age 28 had bilateral ovarian cystectomy and four months later total abdominal hysterectomy and bilateral salpingo oophorectomy for bilateral ovarian MPSC, is reported. She was free of disease for the next 15 years and then presented with a central pelvic mass. At laparotomy, a recurrence in the form of a solitary invasive peritoneal implant was discovered and completely resected. No postoperative adjuvant therapy was given. To date, 16 years after initial diagnosis of MPSC, and one year after detection of recurrence, the patient is alive, well and without disease. Literature data and this case report support the view that MPSC's should be classified separately from both typical serous borderline tumors and invasive carcinomas of the ovary. PMID- 11055487 TI - Camptothecin and mitomycin combination chemotherapy on ovarian clear cell carcinoma with multiple systemic metastases. AB - Ovarian clear cell carcinoma is known for decreased responsiveness to most anticancer drugs, and the prognosis of patients with ovarian clear cell tumors is very poor. We report such a patient with multiple systemic metastases who showed a complete response to combined camptothecin and mitomycin chemotherapy. During therapy, clinical drug-resistance was observed transiently, but response was restored by intermittent administrations of camptothecin. Such combination chemotherapy may be a useful alternative regimen for advanced and/or metastatic ovarian clear cell carcinoma. PMID- 11055488 TI - Physical properties of acetic acid vital in evaluating the cervix for neoplastic changes. AB - INTRODUCTION: In a "developing country" the need exists for an inexpensive, simplistic yet sensitive screening procedure which furthermore, allows for the immediate referral of patients. Research has indicated that the acetic acid test is a most viable option. Screening, however, often occurs in less than ideal conditions thus implying the exposure of the acetic acid to various fluctuating elements. OBJECTIVE: Determining the influence of temperature, time and humidity on the pH of a 5% acetic acid solution. METHOD: The effect of temperature, time and humidity on the pH of a 5% acetic acid solution was determined under controlled laboratory conditions utilizing the calculated molar conductance and dissociation constant. RESULTS: The pH of the 5% acetic acid solution remained a constant throughout exposure to the various elements. DISCUSSION: The results imply that the quality of acetic acid, on which many alternative screening methods and colposcopy are based, remains stable under varying conditions. This is important when screening is implemented in less than ideal conditions utilizing cervicography or the acetic acid test which are primarily dependent on acetic acid. PMID- 11055489 TI - Metastatic carcinoma in a transposed ovary after radical hysterectomy for a stage 1B cervical adenosquamous cell carcinoma. Case report. AB - A 41-year-old multigravida woman presented with a cervical stage 1b adenosquamous cell carcinoma with metastasis in a transposed ovary. Twenty-four months following a radical hysterectomy for cervical cancer a CT scan revealed a mass measuring 7x6x4 cm in size lateral to the ascending colon. The pathological diagnosis showed the mass to be metastatic adenocarcinoma in the transposed ovary which was considered to be adenocarcinoma from the previous cervical adenosquamous cell carcinoma. Thereafter, she was administered a combination of mitomycin C, etoposide and cisplatin (MEP) three times. One year following the completion of the chemotherapy she is still alive without any definitive recurrence. It is important to clarify whether or not ovarian transposition is safe and effective in helping surgeons determine whether or not to preserve the ovarian function for a young patient likely to undergo postoperative irradiation for early invasive cervical cancer. In the present report, we also review the current literature on this subject and discuss our findings. PMID- 11055490 TI - Mullerian adenosarcoma of the uterus: case report and review of literature. AB - Mullerian adenosarcoma--a variant of mullerian mixed mesodermal tumor of the uterus--is typically composed of benign but sometimes mildly atypical glandular epithelial elements admixed with malignant sarcomatous stroma. This rare tumor, which accounts for only about 8% of all uterine sarcomas, usually originates in the endometrium and grows as a polypoid mass within the endometrial cavity. The most prevailing presenting symptom is abnormal vaginal bleeding and the most common finding is a polypoid mass protruding through a dilated cervical canal. The case of a woman, who at age 62 presented with symptoms and signs of acute pelvic inflammatory disease and on vaginal examination an infected mullerian adenosarcoma protruding through a dilated cervical canal was discovered, is reported. Treatment consisted of extensive antibiotic treatment and surgery comprised of total abdominal hysterectomy and bilateral salpingo-oophorectomy followed by postoperative adjuvant pelvic radiotherapy. One year later, the patient is alive with no evidence of disease. PMID- 11055491 TI - Abdominal cystic lymphangioma in a woman at 14 weeks' gestation: case report. AB - Lymphangiomas are uncommon benign tumors that most commonly present early in life, and are even more exceptional in adults. We present a rare case of a 14 week pregnant woman who had a large septated cystic lesion adherent to the ileum. Despite analysis by ultrasound, the correct diagnosis was established only via laparotomy and she had surgical resection of an abdominal lymphangioma. PMID- 11055492 TI - Surgical treatment of invasive carcinoma of the vulva. Our experience. AB - OBJECTIVE: The purpose of this study was to investigate the clinical findings, treatment and outcome of patients with vulvar carcinoma in the L'Aquila area. METHODS: Fifteen cases of vulvar carcinoma seen between September 1991 and December 1999 at the Department of Obstetrics and Gynecology of the University of L'Aquila were reviewed. Clinical, pathologic, surgical and follow-up data were collected from patient records. Mean age at diagnosis was 66.4 years. All patients were evaluated through a careful medical history and physical examination, vulvoscopy, abdomino-pelvic CT or MR, urethrocystoscopy, rectocolonscopy and SCC, and CEA determination. Radical surgery included six patients treated by the Taussig-Way operation. Modified radical surgery accounted for nine patients treated by the Byron three-incision approach. RESULTS: The major early complication was groin wound breakdown which occurred in four cases. The major late complication was chronic leg edema which was reported in six patients. The average number of nodes removed per patient was 19.5. Seven patients (46.7%) had a T2N0M0 pathologic stage, four (26.7%) were T2N1M0, four (26.7%) T1N0M0. Five patients died of local and distant recurrences within 37 months after surgical treatment; ten patients are alive, nine are apparently free from disease whereas one presented local and systemic recurrence within 18 months after surgery. CONCLUSIONS: Vulvar carcinoma predominantly affects older women. Most patients in our series (11/15) had tumors more than 2 cm in diameter. Although the vulva is an external organ and early detection should be achieved, many patients presented with extensive primary lesions due to both patient and physician delay. Stage of disease, tumor size, and nodal metastases are potential prognostic factors useful in selecting patients for a more conservative surgical approach. PMID- 11055493 TI - Ovarian germ cell neoplasm in pregnancy. AB - Ovarian germ cell neoplasm in pregnancy, with variable outcomes after debulking surgery is described. PMID- 11055494 TI - Percutaneous endoscopic gastrostomy (PEG) in palliative treatment of non-operable intestinal obstruction due to gynecologic cancer: a review. AB - Percutaneous endoscopic gastrostomy (PEG) is a relatively simple method in achieving non-surgical gastric decompression in patients with upper gastrointestinal tract obstruction from metastatic pelvic and abdominal tumors. PMID- 11055495 TI - Postchemotherapy late recurrence of non metastatic gestational trophoblastic disease following a partiale mole. A case report. AB - The authors describe a case of a 35-year-old woman who showed elevation of betahCG 13 months after the complete regression of betahCG values following chemotherapy for an incomplete mole. This case outlines the necessity for careful monitoring of betahCG levels in low risk gestational trophoblastic diseases for a period of time longer than one year after achieving the first clinical remission. PMID- 11055496 TI - Estimation of neovascularisation in hyperplasia and carcinoma of endometrium using a "power" angio-Doppler technique. AB - The aim of this study was to estimate neovascularisation in hyperplasia and carcinoma of the endometrium using a "power" angio-Doppler technique. One hundred and eighty-two postmenopausal patients with irregular bleeding from the sexual organs were investigated. Age of the patients ranged between 46 and 78 years. No neovascularisation in a control group of women with normal histopathological results of the endometrium was observed. The sensitivity of this method with reference to the diagnosis of hyperplasia and cancer of the endometrium was 12.2% and 81.2%, respectively. The specificity and positive prediction values were equal to 100% each in both pathologies. The values of the analysed flow indices in the neovascular arteries in endometrial cancer were significantly lower (p<0.05) (properly PI-0.92+/-0.12; RI-0.46+/-0.08) in comparison to the corresponding values in hyperplasia of the endometrium (PI-1.38+/-0.28 and RI 0.66+/-0.18). In conclusion, using a "power" angio-Doppler technique irregular vascularity of the endometrium in a group of patients with hyperplasia was observed in 12.2% of all patients and in 81.2% of those with cancer of the endometrium. The analysis of the values of the blood flow qualitative parameters in the neovascularisation areas in the endometrium at the significance level of PI < or = 1.0; RI < or = 0.5 and TAMV > 18.0 cm can help determine the essential element in the differential diagnosis of benign and malicious lesions. Transvaginal ultrasonography with a "power" angio-Doppler technique can be a valuable diagnostic method in hyperplasia and cancer of the endometrium, especially useful in the early stages of these pathologies. PMID- 11055497 TI - Breast tumors during adolescence. AB - A variety of benign tumors may involve the breast of the adolescent female. Neoplasms and cysts originating from the breast tissue itself, as well as from anatomically related tissues such as lymph nodes may occur. Imaging of the adolescent with a breast mass varies from that of the mature women because of the extremely uncommon occurrence of breast malignancy is this younger population. During a 22-year period (1978-99), 684 young females (14-20 years) were referred to the Breast Unit of the 1st Department of Obstetrics and Gynecology in the major University-appointed hospital of the University of Athens, because of breast masses. The clinical and imaging evaluation of the masses confirmed their presence in the majority of cases. Most cases (442-64.6%) were managed conservatively and carefully followed-up. In 242 cases (35.4%) ablation of the masses was performed. Biopsy showed 236 (97.5%) benign and 6 (2.5%) malignant tumors. The malignant tumors were 2 cases of hemangiosarcoma, 1 case of rhabdomyosarcoma, 1 case of ductual carcinoma, 1 case of cystosarcoma phylloides and 1 case of metastatic rhabdomyosarcoma from the eye. In three cases patients died as a consequence of the disease, two survived and one was lost due to the family's wishes to be transfered to another oncology department abroad. PMID- 11055498 TI - Retinoids in breast cancer prevention and treatment. A review of the literature. AB - During the last three decades, research focused on cancer treatment has led to the development of many cytotoxic agents. Despite the fact that these efforts have significantly improved the prognosis of certain malignancies such as some lymphomas, leukemias and testicular carcinomas, other tumors such as ovarian, lung and metastatic breast cancer are still associated with a poor prognosis. An innovative approach has recently emerged, thanks to a better understanding of tumor cell biology and many efforts are aimed at finding compounds capable of restoring a more differentiated phenotype to tumor cells, thereby reducing the tumor's aggressiveness and ultimately reverting it to its normal counterpart [1, 2]. Retinoids are the prototype of this new therapeutical approach called "differentiation therapy". PMID- 11055499 TI - Clear cell adenocarcinoma of the vulva arising in endometriosis. A case report. AB - A clear cell carcinoma, originating from a focus of endometriosis in the vulva, in a 52-year-old woman, operated on eight years before for ovarian endometriosis is reported. Malignant transformation of extraovarian endometriosis is rare. To date only three cases that originated in the vulva have been reported. PMID- 11055500 TI - Comparison of endometrial pathologies collected by hysteroscopy and hysterectomy in postmenopausal breast cancer tamoxifen-treated patients. AB - PURPOSE OF INVESTIGATION: 1) To assess whether endometrial specimens obtained from removed uteri might show an increase in endometrial pathologies which had been previously diagnosed by hysteroscopy in postmenopausal breast cancer tamoxifen-treated patients. 2) To assess whether hysteroscopy is an efficient method of detecting endometrial pathologies in such patients. METHODS: The findings of two consecutive pathological evaluations in 18 postmenopausal breast cancer tamoxifen-treated patients, performed 11.2+/-11.2 months apart, were compared. The first specimen was collected by hysteroscopy and the second was obtained following hysterectomy. RESULTS: The most significant changes observed were three new cancers diagnosed at hysterectomy, one of which was poorly differentiated. In the first (hysteroscopy) samplings, one patient had atrophic endometrium, a second patient had endometrial proliferation and a third patient had a benign endometrial polyp. Overall, 55.6% of the study patients had various endometrial pathologies in the first sampling, while 83.3% had endometrial pathologies in the second sampling. However, this difference was not statistically significant. CONCLUSION: 1) Endometrial histologic evaluations, performed on removed uteri 11.2+/-11.2 months following previous endometrial samplings of postmenopausal breast cancer tamoxifen-treated patients, showed a non-significant risk of developing overall endometrial pathologies. 2) Hysteroscopy may have missed some endometrial pathologies which were diagnosed later on in specimens obtained by hysterectomy. PMID- 11055501 TI - Serum progesterone, estradiol-17beta and testosterone at the time of relapse in patients with epithelial ovarian cancer. AB - Raised levels of steroid hormones may be detected in women with ovarian cancer at the time of diagnosis. The goal of this study was to investigate the levels of progesterone, testosterone and estradiol-17beta in patients with relapsed epithelial ovarian cancer. We studied 52 patients with a histologic diagnosis of ovarian cancer; 46 of 52 patients were affected by epithelial tumors, two patients had sexcord-stromal tumors, one patient had a germ cell tumor and three patients had a metastatic cancer from the bowel. Of 34 patients with disease relapse, none had elevated serum testosterone levels (>1 ng/ml), one patient (2.9%) had an elevated serum progesterone level (>1.24 ng/ml) and two patients (5.9%) had elevated estradiol-17beta levels (>28 pg/ml). The relationship between the three hormone levels at the time of initial treatment and at relapse was tested using the Students's t-test. At the time of initial treatment venous concentrations of progesterone, estradiol-17beta and testosterone were higher and statistically different (p<0.05) from samples obtained at the time of relapse in the same patients. No significant differences were found between patients studied at the time of relapse and the control group. Measurement of progesterone, estradiol-17beta and testosterone is not helpful in detecting disease relapse in patients with epithelial ovarian cancer. PMID- 11055502 TI - The long term results of radiotherapy with or without surgery in management of advanced vulvar cancer: report of 76 patients. AB - BACKGROUND: The combination of conservative surgery plus radiotherapy for vulvar cancer has been well established as a therapeutic alternative to extensive radical surgery. This study was undertaken to evaluate the long-term results of radiotherapy with or without surgery in the management of advanced vulvar cancer. PATIENTS AND METHODS: The cases of 76 patients who had advanced carcinoma of the vulva treated with different modalities at the University of Texas M.D. Anderson Cancer Center were retrospectively reviewed. Three patients had unstaged disease as a result of previous surgery, 19 had stage II, 40 had stage III, and 14 had stage IV disease. Follow-up ranged from 4 to 17 years (median, 11 years). RESULTS: Five-year disease-free survivals were 75, 67, 68 and 52% for treatment groups I, II, III, and IV, respectively. Disease was controlled locally in 83, 80, 73 and 56% of patients in groups I through IV, respectively; the overall rate of local control was 79%. There was no significant difference in primary tumor control, 5-year disease-free survival, or overall survival among the different treatment groups (p=0.1300). However, these rates did differ significantly (p<0.006) based on FIGO stage of disease. CONCLUSION: In this report, the cure of vulvar cancers with radiotherapy alone (5-year disease-free survival 52% and local control 56%), the radiotherapeutic salvage of patients with surgical failure and/or large tumors, the improved survival with low morbidity by pre- and postoperative radiotherapy were provocative observations suggesting the value of this therapy for advanced vulvar cancer. PMID- 11055503 TI - Uterine adenosarcoma with rhabdomyosarcomatous overgrowth. Brief communication. AB - A case of a rhabdomyosarcomatous uterine adenosarcoma with sarcomatous overgrowth is presented. The rhabdomyosarcoma component constituted about 90% of the tumor. The patient died 5 years and 5 months after the operation. Rhabdomyosarcomatous uterine adenosarcoma even with a sarcomatous overgrowth does not seem to be more malignant than other adenosarcomas. PMID- 11055504 TI - Acute myeloid leukemia: therapeutic indications. AB - The main issue for younger patients with acute myeloid leukemia is the prevention of relapse. About 55% of patients relapse and the risk can partially be predicted by prognostic factors, particularly cytogenetics. A number of strategies can attempt to reduce the relapse risk. Intensification of induction therapy has been attempted but there is as yet no convincing evidence that survival is improved. Transplantation of either allogeneic or autologous stem cells does not seem to offer major survival advantage overall or within risk groups. Improved understanding of resistance mechanisms and the identification of new risk factors may enable the development of a more targeted approach to therapy. PMID- 11055505 TI - Conditioning regimens for allogeneic stem cell transplants. AB - In addition to providing cytoreduction at myeloablative dose intensity, conditioning regimens for allogeneic transplantation are designed to immunosuppress the recipient to permit donor lymphohematopoietic engraftment and thereby establish a graft-versus-malignancy effect. Increased confidence in the potency of this allogeneic graft-versus-malignancy effect, together with the need to reduce dose intensity to make transplantation safer and more widely applicable in older patients, has led to a conceptual revolution in conditioning regimen design. Novel nonmyeloablative transplant conditioning treatments have low regimen-related toxicity and low transplant-related mortality. The transplants confer a graft-versus-malignancy effect in myeloid and lymphoid malignancies and in metastatic renal cell cancer. Future prospects are for low toxicity conditioning regimens combined with specific antileukemia or antitumor intensification with radioconjugated or unmodified antibodies and the application of highly immunosuppressive but low toxicity conditioning regimens for mismatched transplants. PMID- 11055506 TI - Bone marrow or peripheral blood as a source of stem cells for allogeneic transplants. AB - Peripheral blood stem cell transplants are being increasingly used in the allogeneic setting and are often preferred to the conventional bone marrow source. The aim of this report is to review available data on peripheral blood versus bone marrow hematopoietic stem cell transplantation. The discussion is restricted to HLA-identical sibling transplants receiving unmanipulated grafts. This is because data with appropriate follow-up are available only for this type of comparison: we have preliminary data on the use of peripheral blood from unrelated donors, and on the use of T-cell depletion/CD34+ selection methods. The latter are evolving rapidly and it may be difficult to find a concurrent group of patients receiving T-cell-depleted or CD34-selected marrow. The results of retrospective and prospective studies are similar: hematologic and immune recovery are faster after peripheral blood grafts, acute graft-versus-host disease is comparable, whereas chronic graft-versus-host disease is increased in recipients of peripheral blood transplants. Transplant-related mortality is similar in the two groups, whereas disease recurrence is lower after peripheral blood grafts. The general opinion is that peripheral blood grafts are indicated for patients with advanced disease, whereas for patients with early-phase disease the two sources may give comparable results. PMID- 11055507 TI - Developments in T-cell depletion of allogeneic stem cell grafts. AB - The quantitative depletion of T-cells has long been established as a highly effective way to prevent graft-versus-host disease. The disadvantages associated with T-cell depletion (TCD), including loss of the graft-versus-leukemia effect and impaired immune recovery have been important drawbacks for application on a wider scale. Recently, however, new approaches such as the use of peripheral blood stem cells and the application of TCD, in conjunction with delayed donor lymphocyte infusion, have opened new possibilities. The first clinical studies addressing the merits of these approaches suggest that effective prevention of graft-versus-host disease may be achieved by TCD, whereas the graft-versus leukemia effect may be maintained by delayed donor lymphocyte infusion. Also, immune recovery may be accelerated by the use of peripheral blood stem cells. New techniques to manipulate stem cell grafts in vitro are being developed with the purpose to either selectively delete alloantigen specific T cells or tolerize such T cells by blocking costimulatory pathways. As a result, the in vitro manipulation of T cells of allogeneic stem cell grafts currently enjoys renewed scientific interest. PMID- 11055508 TI - Umbilical cord blood transplants. AB - With the establishment of cord blood banks, the number of related and unrelated umbilical cord blood transplants is increasing worldwide. Close links have been established with the cord blood banks. Available data showed that umbilical cord blood transplants offer overall results comparable to those obtained with related or unrelated bone marrow transplants. Several differences were found: engraftment with cord blood was delayed, resulting in an increased incidence of early transplant complications, and the incidence of acute and chronic graft-versus host disease was significantly reduced with cord blood grafts, even in HLA mismatched transplants and in adults. In patients with leukemia, the rate of relapse appeared to be similar to that documented in bone marrow transplant recipients. These data confirm the potential benefit of using umbilical cord blood hematopoietic stem cells for allogeneic transplants. PMID- 11055509 TI - Role of mesenchymal stem cells in hematopoietic stem cell transplantation. AB - Within the bone marrow stroma are multipotential cells which are capable of differentiation into a number of mesenchymal cell lineages. These cells, termed mesenchymal stem cells, have recently been identified and characterized in humans. Many studies indicate that the bone marrow stroma is damaged following bone marrow transplantation. Since the marrow stroma is critical for the maintenance of hematopoiesis, its ability to support hematopoiesis following stem cell transplantation may be impaired. Animal models suggest that the transplantation of healthy stromal elements, including mesenchymal stem cells, may enhance the ability of the bone marrow microenvironment to support hematopoiesis after stem cell transplantation. Here the authors review recent data that suggest that mesenchymal stem cells may possess therapeutic value not only for the repair of damaged mesenchymal tissues following hematopoietic stem cell transplantation, but also as potential vectors for the delivery of corrective genes. PMID- 11055510 TI - Issues in the manufacture and transplantation of genetically modified hematopoietic stem cells. AB - The advent of safe and practical means to correct, enhance or protect blood cells at the genetic level offers tantalizing therapeutic perspectives. At present, gene delivery using a replication-defective retrovirus is the most efficient method to stably transduce hematopoietic cells. The successful adaptation of retroviral infection to hematopoietic stem cells requires optimized transduction conditions that maximize gene transfer while preserving the cells' potential for engraftment and longterm hematopoiesis. The successful establishment of effective transduction protocols hinges on retrovirus biology as well as stem cell and transplantation biology. Interestingly, the genetic approach could permit novel strategies to promote host repopulation by transplanted stem cells. However, regulated and predictable expression of any transgene integrated at random chromosomal locations cannot be taken for granted. Investigation of the control of transgene expression and prevention of vector silencing will become increasingly important. PMID- 11055511 TI - Increasing transfusion safety by reducing human error. AB - Transfusion of the wrong blood is a rare but measurable event that may result in serious complications and whose main cause is human error. Any preventive strategy should be based on a careful assessment of the incidence of these events and of their causes, and requires a standardized confidential reporting system, to avoid underreporting, covering also near misses. Creating or revising written procedures and monitoring their implementation are indispensable to improve blood safety, but human error can occur in spite of these measures. Technologic instruments are now available to fill the gap between written and implemented procedures, forcing the operator to carry out the critical steps in the process according to the adopted guidelines. PMID- 11055512 TI - Nucleic acid testing for transfusion-transmissible agents. AB - Pooled nucleic acid testing for HIV and hepatitis C virus (HCV) has been successfully implemented in the United States. Reactive rates for nucleic acid testing (NAT) are comparable to those for serological testing. The combined yield for HIV and HCV RNA confirmed-positive, seronegative donations for the 1st year of testing has been 62 HCV NAT-reactive donations among more than 16.3 million screened (1:263,000) and four HIV NAT-reactive, p24 antigen-negative donations among greater than 12.6 million screened (1:13,150,000). The observed yield of NAT confirmed-positive donations is compatible with estimates derived from window period projections and recent measurements of incidence. Of the HCV NAT-reactive donations identified, one has not sero-converted over a 300-day period. The predominant recent risk factor identified for HCV NAT-reactive donors is intravenous drug use. PMID- 11055513 TI - Hepatitis C lookback. AB - Hepatitis C was responsible for the majority of cases of posttransfusion hepatitis before the introduction of a specific screening test for blood donors. Infected recipients may remain asymptomatic for many years, but cirrhosis and hepatocellular carcinoma may develop decades after infection. Lookback, or the identification of recipients of potentially contaminated blood, is now being conducted in many countries, including Holland, Denmark, the United Kingdom, Canada, New Zealand, and the United States. In targeted lookback, recipients of blood from donors subsequently found to be positive for hepatitis C are notified and advised to undergo testing. In general lookback, all patients who received blood before being tested for hepatitis C are advised to undergo testing. Difficulties with both forms of lookback illustrate the importance of vein-to vein tracking of blood products, including the potential utility of a centralized registry of blood product recipients. PMID- 11055514 TI - Universal leukocyte reduction. AB - Leukocyte reduction of blood components, in the United States, is generally reserved for conditions in which a clinical indication has been documented. There is no evidence that either Creutzfeldt-Jakob disease or variant Creutzfeldt-Jakob disease are transmitted by transfusion in humans or that leukocyte reduction of blood components could reduce their transmission. A number of adverse outcomes following transfusion are alleged to be the result of white blood cells. At this point in time, there are insufficient clinical data to justify the universal leukocyte reduction of blood components. PMID- 11055515 TI - Transfusion-related acute lung injury. AB - Transfusion-related acute lung injury is a life-threatening complication of hemotherapy associated with the transfusion of plasma-containing blood products. It is characterized by acute respiratory distress, pulmonary edema and hypoxemia. Although its frequency is unknown, Food and Drug Administration data suggest that it is the third most common cause of transfusion-associated deaths, representing 9% of reported cases. Males and females of all ages are at equal risk. To date, there is no recognized profile of individuals who are at increased risk for this complication. Although there are two purported mechanisms of injury, the preponderance of evidence suggests that passively transfused complement activating antibodies (either granulocyte or HLA-specific) act as mediators, which result in granulocyte aggregation, activation, and microvascular pulmonary injury. With appropriate respiratory intervention, most patients recover within 96 hours of the original insult and without permanent pulmonary sequelae. PMID- 11055516 TI - New products for managing inhibitors to coagulation factors: a focus on recombinant factor VIIa concentrate. AB - The treatment of alloantibody and autoantibody inhibitors directed against the factor VIII coagulant protein is one of the most challenging and expensive problems in hematology. Because the currently available plasma replacement products used in this context do not control the bleeding complications in all patients, and because of the usual emergent quality of the bleeding complications, there has been a definite need to have a uniformly reliable product for instant use, which possesses a high degree of hemostatic reliability and safety. The recent introduction of recombinant factor VIIa (rFVIIa) has been a welcome addition to the pharmacologic armamentarium for the treatment of neutralizing antibodies against coagulation factors. The mechanisms of action of rFVIIa have also been interesting and have provided insight into how the coagulation pathway accomplishes adequate hemostasis. This review will discuss this new medication and place into the context of coagulation inhibitor therapy. PMID- 11055517 TI - Laboratory diagnosis of autoimmune cytopenias. AB - The past year's literature shows that little progress has been achieved in the laboratory diagnosis of autoimmune hemolytic anemia. The direct antiglobulin test is the only diagnostic test for autoimmune hemolytic anemia. Advantages of new techniques, such as the gel test, have to be determined. Today, cephalosporins are known to cause both drug-dependent and -independent autoantibodies. The diagnosis of idiopathic thrombocytopenic purpura is a clinical one. The new assays that measure antibodies against specific glycoproteins offer improved specificity. New laboratory advancements and accumulation of data on granulocytes' antigens and antibodies enabled us to recommend guidelines for the laboratory investigation of autoimmune neutropenia. PMID- 11055519 TI - Current world literature. Hematopoietic stem cell transplantation. PMID- 11055518 TI - Cost-effectiveness analysis and the selection of blood products. AB - A new generation of blood components and transfusion modalities aimed at further reducing real or hypothetical risks threatening the blood supply is available. Since the risk of infection with transfusion-transmitted viruses is already very low, many of these new interventions provide a minimal health benefit at a very high cost. It is even possible that risks inherent to the new technologies themselves could surpass the current risk of transfusion-transmitted viruses. For some of the new blood components, such as leukoreduced products, the poor cost effectiveness projections may be substantially improved if the purported health benefits are eventually proved in appropriate randomized, controlled clinical trials. Meanwhile, cost-effectiveness analyses show that allocating more resources to improve the safety of donated blood may actually decrease the overall transfusion safety by subtracting resources from other areas of the health care system, so a realignment of costs with health benefits seems necessary. PMID- 11055520 TI - Current world literature. Transfusion medicine. PMID- 11055521 TI - Mental illness, criminality, and citizenship. PMID- 11055522 TI - From dangerousness to risk assessment of community violence: taking stock at the turn of the century. AB - This article focuses on a history of ideas-the history of the last quarter century, in which great advances have occurred in the reshaping of the concept of dangerousness while, at the same time, disappointingly little has happened to improve the abilities of frontline clinical decision-makers to make violence risk assessments. We are now operating in a fundamentally changed environment in the three core areas of research, clinical practice, and law as a result of evolving thinking and new data, shown in the following ways. (1) Risk assessment concepts and associated research models have altered the focus to probabilistic thinking and ideas of graduated interventions as opposed to the Yes-No, In-Out dichotomies formerly associated with the concept of dangerousness. (2) Likewise, it has become clear that dangerousness is a legal and not a medical concept, in which the actual clinical corollary is risk of future violent behavior. (3) The accuracy of psychiatric assessments of future violent behavior is limited, but it may increase for specific subgroups within specified time frames and locations. PMID- 11055523 TI - Commentary: assessing the risk of violence--are "accurate" predictions useful? PMID- 11055524 TI - Commentary: dangerousness--a failed paradigm for clinical practice and service delivery. PMID- 11055525 TI - Forensic psychiatrists' fee agreements: a preliminary empirical survey and discussion. AB - The author performed a preliminary empirical study of forensic fee agreements. A survey of forensic psychiatrists produced samples of fee agreements used for expert witness work. The samples were analyzed to determine patterns of usage, critical elements, styles of agreement design, and other observations. Despite uncertainties of payment in this field, a surprising number of experts do not use formal fee agreements. This and other results are discussed. PMID- 11055526 TI - Interpreting clinical evidence of malingering: a Bayesian perspective. AB - Customary ways of reporting on or testifying about malingering have shortcomings. Stating an opinion "with reasonable medical certainty" tells fact-finders little about how much confidence the opinion deserves; stating that an individual's behavior is similar to that of known malingerers does not convey the information that fact-finders really need to know, which is the likelihood that the evaluee in question is a malingerer, given the evaluator's findings. Mossman and Hart (Mossman D, Hart KJ: Presenting evidence of malingering to courts: insights from decision theory. Behav. Sci. Law 14:271-91, 1996) recommend that mental health professionals address this problem by using Bayes' theorem to interpret test data from evaluations. However, these authors do not discuss the use of evidence obtained during interviews and from other clinical contexts, nor do they describe a method for quantifying imprecision in Bayesian probabilities. This article provides examples of how forensic evaluators might use a Bayesian perspective to interpret clinical indicia of malingering observed during evaluations of adjudicatory competence. The article discusses sources of imprecision in Bayesian posterior probabilities, describes a method for characterizing that imprecision using confidence intervals, and then presents several sample calculations that illustrate how interview findings change the likelihood of malingering. The article also discusses the implications of the Bayesian approach for forensic evaluations and for future research on malingered incompetence. PMID- 11055527 TI - The relationship of deviant sexual arousal and psychopathy in incest offenders, extrafamilial child molesters, and rapists. AB - The relationship between deviant sexual arousal, as measured by auditory phallometric stimuli, and psychopathy, as measured by the Psychopathy Checklist Revised, was examined in 156 incest offenders, 260 extrafamilial child molesters, and 123 rapists. Subjects in each group had never been convicted of another type of sexual offense. Replicating previous research, rapists were more psychopathic than incest offenders and child molesters. Deviant sexual arousal to auditory stimuli was evident only on the Pedophile Index for child molesters. When the relationship between psychopathy and deviant sexual arousal was evaluated in the three groups combined, several significant correlations emerged. However, a finer analysis of these correlations revealed that child molesters evidenced a significant correlation between psychopathy and the Rape Index and psychopathy and the Pedophile Index. There were no such significant findings in the incest offender or rapist groups. Implications of the results are discussed. PMID- 11055528 TI - The effect of psychopathy on outcome in high security patients. AB - The study aimed to examine the relationship between the total score on Hare's revised Psychopathy Checklist (PCL-R) and aspects of outcome for a nonrandom sample (n = 89) of male mentally disordered offenders treated in an English high security hospital. The subjects were all legally classified as suffering from "psychopathic disorder" and the majority were followed-up in the community. The methodology was retrospective, using existing case-file data, with follow-up lasting until discharge from statutory supervision. PCL-R scores were dichotomized and related to various outcome factors, including recidivism and aspects of social behavior. The results showed, in contrast to previous North American research, that the PCL-R did not predict any of the outcome factors. Because the PCL-R was able to identify psychopaths in this population but failed to predict their prognosis, it is possible that their outcome may have been improved by the treatment they received in hospital. PMID- 11055529 TI - Criminal profiling: is there a role for the forensic psychiatrist? AB - Criminal profiling is a field that has gained notoriety in the mainstream consciousness, yet few people realize what it is that criminal profilers actually do and who is doing it. Suffering from a limited applied scientific literature that seems overshadowed by memoir trade books and journalistic style research, the field lacks a consensus regarding required expertise, ethics, methods of profiling, and research needs. This would seem to beg the question, why would anyone turn to a criminal profiler? After all, what would a profiler have to offer to a police investigator? This article will examine criminal profiling from the viewpoint of what it is, what it should be, and whether or not the forensic psychiatrist has a role to play in this field. The author will also argue that, of the available profiling methods, the deductive method is best suited to the training and expertise of the forensic psychiatrist. PMID- 11055531 TI - A community policing approach to persons with mental illness. PMID- 11055532 TI - Improving high risk encounters between people with mental illness and the police. PMID- 11055533 TI - Police response to mental health emergencies--barriers to change. PMID- 11055534 TI - Policing the emotionally disturbed. PMID- 11055535 TI - Davis v. Monroe County Board of Education et al. PMID- 11055536 TI - America's daughters on Gandhi's daughters. PMID- 11055537 TI - Rape, justice, and hierarchy in India. PMID- 11055538 TI - The future of forensic psychiatry. PMID- 11055539 TI - The future of forensic psychiatry. PMID- 11055540 TI - Wendell Williamson v. Dr Myron Liptzin case. PMID- 11055541 TI - Insulin does not regulate the promoter of cholesteryl ester transfer protein (CETP) in HIRc/pCETP-CAT cells. AB - Cholesteryl ester transfer protein (CETP) is a plasma enzyme involved in cholesterol metabolism. As a potential target in the treatment of atherosclerosis, a number of studies have focused how this enzyme is regulated. It has been postulated that insulin may regulate CETP gene expression, and these effects may be mediated through CCAAT/enhancer binding protein alpha (C/EBPalpha). The present study examines the effects of insulin on the activity of the CETP promoter in rat fibroblasts expressing the human insulin receptor (HIRc). HIRc cells were stably transfected with a chimeric construct containing 3.2 kb of the CETP promoter attached to the bacterial chloramphenicol acyltransferase gene (pCETP-CAT) without significantly affecting the expression of the insulin receptor. CAT activity was 8-fold higher in cultured HIRc/pCETP CAT in the presence of 100 mg/dL LDL cholesterol, than those cultured without cholesterol (p < 0.05). However, culturing these cells in the presence of 100 nM insulin did not result in any change in CAT activity when compared to control cells. In HIRc/pCETP-CAT cells transiently transfected with a construct that constitutively expressed C/EBPalpha protein, a 3-fold increase in CAT activity was observed when compared to cells transiently transfected with non-specific DNA (p < 0.05). However, no observable effect on the CETP promoter was observed in the presence of insulin. Thus, in HIRc/pCETP-CAT cells, we were unable to substantiate the hypothesis that insulin regulates CETP gene transcription. These results suggest that the effects of insulin on CETP expression regulation may be downstream of transcription. PMID- 11055542 TI - Intracellular reduction of selenite into glutathione peroxidase. Evidence for involvement of NADPH and not glutathione as the reductant. AB - Selenium (Se) in selenite is present in an oxidized state, and must be reduced for it to be incorporated as selenocysteine into selenoenzymes such as glutathione peroxidase (GPx). In vitro, Se, as in selenite, can be reduced utilizing glutathione (GSH) and glutathione reductase (GRed). We determined the effects of decreasing GSH levels, inhibiting GRed activity, and decreasing cellular NADPH on the selenite-dependent rate of GPx synthesis in cultured cells: PC3, CHO, and the E89 glucose-6-phosphate dehydrogenase (G-6-PD)-deficient cell line. A novel statistical analysis method was developed (using Box Cox transformed regression and a bootstrap method) in order to assess the effects of these manipulations singly and in combinations. Buthionine sulfoximine (BSO) was used to decrease GSH levels, 1,3 bis-(2 chloroethyl)-1 -nitrosourea (BCNU) was used to inhibit GRed activity and methylene blue (MB) was used to decrease cellular NADPH levels. This statistical method evaluates the effects of BSO, BCNU, MB and selenite alone and in combinations on GPx activity. Decreasing the GSH level (< 5% of control) did not have an effect on the selenite-dependent rate of GPx synthesis in PC3 or CHO cells, but did have a small inhibitory effect on the rate of GPx synthesis in E89 cells. Inhibiting GRed activity was also associated with either no effect (CHO, E89) or a small effect (PC3) on GPx activity. In contrast, decreasing NADPH levels in cells treated with MB was associated with a large decrease in the selenite-dependent rate of GPx synthesis to 36, 34 and 25% of control in PC3, CHO, and E89 cells, respectively. The effects of BSO plus BCNU were not synergistic in any of the cell lines. The effects of BSO plus MB were synergistic in G-6-PD-deficient E89 cells, but not in PC3 or CHO cells. We therefore conclude that under normal culture conditions, NADPH, and not glutathione, is the primary reductant of Se in selenite to forms that are eventually incorporated into GPx. For cells with abnormal ability to generate NADPH, lowering the GSH levels had a small effect on selenite-dependent GPx synthesis. GRed activity is not required for the selenite-dependent synthesis of GPx. PMID- 11055543 TI - Unmodified calcium concentrations in tumour necrosis factor receptor subtype mediated apoptotic cell death. AB - Tumour necrosis factor-alpha (TNF) receptors mediate a variety of effects dependent on cell type. A role for Ca2+ in TNF-induced death remains uncertain. Here we investigated restricting intracellular/extracellular Ca2+ in HeLa epithelial carcinoma cells expressing low and high levels of p75TNFR receptor subtype and KYM-1 rhabdomyosarcoma cells, models of rapid TNF-induced apoptosis. Ca2+ -chelators EGTA and BAPTA-AM as well as microsomal Ca2+ -ATPase inhibitor thapsigargin, did not alter TNF-induced death. TNF was also unable to alter resting [Ca2+]i levels which remained < 200 nM even during times when these cells were undergoing apoptotic cell death. These findings indicate no role for modulated Ca2+ concentrations in TNF-induced apoptotic cell death. PMID- 11055544 TI - Substitution of two insulin receptor carboxy-terminal tyrosines with phenylalanine impairs the expression of MAP kinase phosphatase-1 (MKP-1) mRNA. AB - Cells expressing mutant insulin receptors (Y/F2), in which tyrosines 1316 and 1322 have been replaced with phenylalanine, exhibit enhanced insulin-induced MAP kinase activity and DNA synthesis in comparison with cells expressing wild type insulin receptors (Hirc B). To elucidate the mechanism of enhanced responsiveness, the expression of MAP kinase phosphatase-1 (MKP-1), a negative regulator of MAP kinase activity, was measured in Hirc B and Y/F2 cells incubated in the absence and presence of insulin for various periods of time, and over increasing concentrations of the ligand. Treatment of both cell lines with insulin induced a time and concentration-dependent relative increase in MKP-1 mRNA expression. However, in Y/F2 cells both basal and insulin-stimulated MKP-1 mRNA levels were more than 60% lower than that observed in cells transfected with the wildtype receptors. Cyclic AMP analog (8-Br-cAMP)/inducer (Forskoline) increased MKP-1 mRNA levels in both cell lines, and to a lesser extent in Y/F2 cells. In contrast to insulin the relative increase in MKP-1 mRNA expression induced by 8-Br-cAMP or forskoline was similar in Y/F2 and Hirc B cells. The overexpression of MKP-1 in Y/F2 cells inhibited insulin stimulated DNA synthesis. Transfection of wild type insulin receptors into Y/F2 cells increased basal levels of MKP-1. These results suggest that insulin receptor tyrosine residues 13/16 and 1322 play an important role in the regulation of MKP-1 expression both under basal and insulin stimulated conditions, and are not necessary for the induction of MKP-1 mRNA by cAMP. Furthermore, the enhanced insulin induced mitogenic signaling seen in Y/F2 cells is, at least in part, due to impaired MKP 1 expression. PMID- 11055545 TI - Selective inhibition of the non-enzymatic lipid peroxidation of phosphatidylserine in rod outer segments by alpha-tocopherol. AB - In the present study it was investigated if alpha-tocopherol shows protection against in vitro lipid peroxidation of phospholipids located in rod outer segment membranes (ROS). After incubation of ROS in an ascorbate-Fe2+ system, at 37 degrees C during 160 min, the total cpm originated from light emission (chemiluminescence) was found to be lower in those membranes incubated in the presence of alpha-tocopherol. The fatty acid composition of total lipids isolated from rod outer segment membranes was substantially modified when subjected to non enzymatic lipid peroxidation with a considerable decrease of docosahexaenoic acid (22:6 n-3). The incorporation of alpha-tocopherol (0.35 micromol/mg protein) produce a 43.37% inhibition of the lipid peroxidation process evaluated as chemiluminescence (total cpm originated in 160 min). The phospholipid species containing the highest amount of docosahexaenoic acid: phosphatidylethanolamine and phosphatidylserine were more affected than phosphatidylcholine during the lipid peroxidation process. Not all phospholipids, however, were equally protected after the addition of alpha-tocopherol to the incubation medium. Phosphatidylcholine and phosphatidylethanolamine, were not protected by alpha tocopherol, the vitamin provides selective antioxidant protection only for phosphatidylserine. These results indicate that alpha-tocopherol may act as antioxidant protecting rod outer segment membranes from deleterious effect by a selective mechanism that diminishes the loss of docosahexaenoic acid from phosphatidylserine. PMID- 11055546 TI - Alterations in binding characteristics of peripheral benzodiazepine receptors in testes by vitamin A deficiency in guinea pigs. AB - The correlation of vitamin A with the binding characteristics of peripheral benzodiazepine receptors (PBRs) in testes have been implicated on the basis of findings of involvement of vitamin A in testicular physiology and the abundance of PBRs in testicular tissue. Both vitamin A and PBRs are involved in the control of cell proliferation and differentiation but no data exists regarding the relationship between them. In the present study, we have examined the effects of vitamin A deficiency on the affinity and density of PBRs in testes of guinea pigs. Weanling guinea pigs were divided into three groups: control, pair-fed control and vitamin A deficient. They were fed a complete semipurified diet. The vitamin A deficient diet was similar to the control diet except vitamin A palmitate was omitted. Vitamin A deficiency status was achieved after 90 days of feeding. Binding of [3H]Ro 5-4864, a specific ligand for peripheral benzodiazepine receptors was determined in whole homogenate of testicular tissue. There was a 77% decrease in the receptor density (B max) in vitamin A deficient group compared to control. The Bmax values for control, pair-fed control and vitamin A deficient groups were: 12.4 +/- 0.4, 8.8 +/- 0.2 and 3.0 +/- 0.6 pmol/g, respectively. The equilibrium dissociation constant (K(D)) values were also 86% decreased in the vitamin A deficient group compared to the other groups. The K(D) values for control, pair-fed control and vitamin A deficient groups were: 3.4 +/- 0.7, 2.8 +/- 0.5 and 0.5 +/- 0.01, respectively. The decrease in the binding characteristics of PBRs in testes due to vitamin A deficiency was accompanied with a corresponding decrease in the levels of testosterone in plasma. These results suggest a close functional relationship of vitamin A with PBRs in testes. PMID- 11055547 TI - Relationship between free radicals and adenosine in the mechanism of preconditioning: are they interrelated or independent triggers? AB - Both free radicals (FRs) and adenosine receptor activation contribute to triggering a mechanism of preconditioning (PC) against infarction. This study examined the possibility that there is some interaction between FRs and adenosine generation during PC. In the first series of experiments, the effects of an FR scavenger, N-2-mercaptopropionyl glycine (MPG), on the interstitial adenosine level during PC and on the infarct size-limiting effect of PC were assessed in the rabbit heart in situ. PC with 5-min ischemia/5-min reperfusion limited infarct size after 30-min coronary occlusion (expressed as a percentage of area at risk, %IS/AR) from 33.2 +/- 4.7% (S.E.) to 10.8 +/- 1.1% (p < 0.05). This cardioprotection was blocked by MPG (1.5 mg/kg/min i.v.) infused before and during PC (%IS/AR = 27.4 +/- 3.6). However, the same dose of MPG did not suppress elevation of the adenosine and inosine levels in the microdialysate from the myocardium during 5-min ischemia/reperfusion. In the second series of experiments, the effect of an FR-generating system (1 mM hypoxanthine and 20 mU/ml xanthine oxidase) on the purine production was compared to that of PC in isolated rabbit hearts. Whereas PC increased the adenosine level in the coronary effluent from 0.17 +/- 0.16 microM under baseline to 1.68 +/- 0.53 microM, infusion of the FR generators over a period of 5 min did not increase the adenosine release. However, infarct size was similarly reduced by PC and by 5-min transient infusion of FR generators, and the cardioprotection by the FR generators was abolished by 300 microM MPG. These results suggest that there is no interaction between free radicals and adenosine during the trigger phase of PC in the rabbit heart. PMID- 11055548 TI - Dexamethasone increases the incorporation of [3H]serine into phosphatidylserine and the activity of serine base exchange enzyme in mouse thymocytes: a possible relation between serine base exchange enzyme and apoptosis. AB - The exposure of phosphatidylserine toward the external surface of the membrane is a well-established event of programmed cell death. The possibility that an apoptotic stimulus influences the metabolism of this phospholipid could be relevant not only in relation to the previously mentioned event but also in relation to the capability of membrane phosphatidylserine to influence PKC activity. The present investigation demonstrates that treatment of mouse thymocytes with the apoptotic stimulus dexamethasone, enhances the incorporation of [3H]serine into phosphatidylserine. Cell treatment with dexamethasone also enhanced the activity of serine base exchange enzyme, assayed in thymocyte lysate. Both the effects were observed at periods of treatment preceding DNA fragmentation. The addition of unlabelled ethanolamine, together with [3H]serine to the medium containing dexamethasone-treated thymocytes lowered the radioactivity into phosphatidylserine. Serine base exchange enzyme activity was influenced by the procedure used to prepare thymocyte lysate and was lowered by the addition of fluoroaluminate, that is widely used as a G-protein activator. The increase of serine base exchange enzyme activity induced by dexamethasone treatment was observed independently by the procedure used to prepare cell lysate and by the presence or absence of fluoroaluminate. PMID- 11055549 TI - Garlic ameliorates hyperlipidemia in chronic aminonucleoside nephrosis. AB - Nephrotic syndrome (NS) is characterized by proteinuria, oxidative stress and endogenous hyperlipidemia. Hyperlipidemia and oxidative stress may be involved in coronary heart disease and the progression of renal damage in these patients. Garlic has been suggested to be beneficial in various disease states. Some of the beneficial effects of garlic may be secondary to its hypolipidemic and antioxidant properties. Therefore, the effect of a 2% garlic diet on acute and chronic experimental NS induced by puromycin aminonucleoside (PAN) was studied in this work. Acute NS was induced by a single injection of PAN to rats which were sacrificed 10 days later. Chronic NS was induced by repeated injections of PAN to rats which were sacrificed 84 days after the first injection. Garlic treatment was unable to modify proteinuria in either acute or chronic NS, and hypercholesterolemia and hypertriglyceridemia in acute NS. However, garlic treatment diminished significantly total-cholesterol, LDL-cholesterol and triglycerides, but not HDL-cholesterol in chronic NS. Garlic induced no change in the percentage of sclerotic glomeruli in chronic NS and a significative decrease on the percentage of sclerotic area of these glomeruli (33 +/- 3% in NS+Garlic group vs. 47 +/- 4% in NS group, p = 0.0126). The enhanced in vivo renal H2O2 production and the diminished renal Cu, Zn-SOD and catalase activities in acute NS, and the decreased renal catalase activity in chronic NS were not prevented by garlic treatment. These data indicate that garlic treatment ameliorates hyperlipidemia and renal damage in chronic NS which is unrelated to proteinuria or antioxidant enzymes. PMID- 11055550 TI - Na+ -transport modulation induces isoform-specific expression of Na+,K+ -Atpase alpha-subunit isoforms in C2C12 skeletal muscle cell. AB - Changes in demands for Na+ transport alter expression of the Na+,K+ -ATPase subunit isoforms. In skeletal muscle, the effects of these changes on expression the alpha2 isoform, the major isoform expressed in differentiated muscle cell, is not known. Therefore, this study examines regulation of the alpha-subunit isoforms by Na+ in the C2C12 skeletal muscle cell that expresses the alpha1 and alpha2 isoforms. Western blot analysis showed that in differentiating C2C12 muscle cell, but not in undifferentiated myoblast, veratridine, a Na+ channel activator, greatly increased expression of the alpha2 isoform; expression of alpha1 was unaltered. Because the level of alpha-actinin was unaltered, the data suggest that veratridine treatment did not significantly alter the progression of cell differentiation. Furthermore, a reduction in Na+ transport by tetrodotoxin again failed to alter expression of alpha1. Thus, in C2C12 skeletal muscle cell, changes in Na+ transport alters expression of the alpha2, but not the alpha1 isoform. These results differ from those observed previously in muscle cells that express only the alpha1 isoform. Because mammalian skeletal muscle expresses both the alpha1- and alpha2-subunit isoforms, the differential regulation that was observed may be physiologically relevant in these muscle cells in vivo. PMID- 11055551 TI - Mouse retinol binding protein gene: cloning, expression and regulation by retinoic acid. AB - A full-length cDNA clone encoding the retinol binding protein (RBP) was isolated from a mouse liver cDNA library by hybridization screening. The nucleotide sequence of murine RBP is 85 and 95% homologous to that of human and rat RBP, respectively, with a deduced amino acid sequence > or = 83% homologous to both species. Analysis of the tissue expression pattern of RBP mRNA in the female mouse indicated relatively abundant expression in the liver, with lesser amounts in extrahepatic tissues including adipose, kidney, spleen and uterus, suggesting that these tissues may have a significant role in retinol homeostasis. Mouse liver cell RBP regulation by retinoids was also investigated. Both all-trans retinoic acid (AT-RA) and 9-cis retinoic acid (9c-RA) induced RBP mRNA expression in a dose- and time-dependent manner. Maximal levels (up to 4-fold above controls) were observed at > or = 48 h following treatment of both mouse hepatoma cells in vitro and in vivo in mice receiving a single, oral dose of either retinoid. Interestingly, 9c-RA was more potent at RBP induction in both in vivo and in vitro systems. Given the extent and temporal pattern of RBP induction, we suggest that the RA-mediated increase in liver RBP is part of a cellular protection mechanism. Increased levels of RBP would facilitate sequestration and possibly cellular export of RA in cells receiving prolonged exposure to high levels of RA, thus minimizing toxicity. PMID- 11055552 TI - Interaction between acylphosphatase and SERCA in SH-SY5Y cells. AB - Ca2+ transport by sarco/endoplasmic reticulum, tightly coupled with the enzymatic activity of Ca2+ -dependent ATPase, controls the cell cycle through the regulation of genes operating in the critical G, to S checkpoint. Experimental studies demonstrated that acylphosphatase actively hydrolyses the phosphorylated intermediate of sarco/endoplasmic reticulum calcium ATPase (SERCA) and therefore enhances the activity of Ca2+ pump. In this study we found that SH-SY5Y neuroblastoma cell division was blocked by entry into a quiescent G0-like state by thapsigargin, a high specific SERCA inhibitor, highlighting the regulatory role of SERCA in cell cycle progression. Addition of physiological amounts of acylphosphatase to SY5Y membranes resulted in a significant increase in the rate of ATP hydrolysis of SERCA. In synchronized cells a concomitant variation of the level of acylphosphatase isoenzymes opposite to that of intracellular free calcium during the G1 and S phases occurs. Particularly, during G1 phase progression the isoenzymes content declined steadily and hit the lowest level after 6 h from G0 to G1 transition with a concomitant significant increase of calcium levels. No changes in free calcium and acylphosphatase levels upon thapsigargin inhibition were observed. Moreover, a specific binding between acylphosphatase and SERCA was demonstrated. No significant change in SERCA-2 expression was found. These findings suggest that the hydrolytic activity of acylphosphatase increase the turnover of the phosphoenzyme intermediate with the consequences of an enhanced efficiency of calcium transport across endoplasmic reticulum and a subsequent decrease in cytoplasmic calcium levels. A hypothesis about the modulation of SERCA activity by acylphosphatase during cell cycle in SY5Y cells in discussed. PMID- 11055553 TI - The effect of thioacetamide on the activity and expression of cytosolic rat liver glutathione-S-transferase. AB - The effect of thioacetamide (TA), an hepatotoxic and hepatocarcinogenic compound, on the expression and activity of the cytosolic enzyme glutathione-S-transferase (GST) was studied in rat liver. Four h following the administration of 14C labeled thioacetamide (50 mg/Kg), several subunits of GST were found to be radioactively labeled. A single sublethal dose of TA (250 mg/Kg) decreased by three-fold the expression of class alpha GST at 24-48 h of treatment, but did not significantly affect the transcription of class mu GST. The activity of the enzyme toward 1-chloro-2,4-dinitrobenzene was mildly inhibited (66% of the control) by a 24 h TA treatment and gradually increased thereafter. It is proposed that the covalent binding of TA or its derivative to the GST subunits does not affect the activity of the enzyme. Nevertheless, GST activity inhibition is due to the deleterious effect of TA on GST transcription. PMID- 11055554 TI - Mechanisms whereby glucose deprivation triggers metabolic preconditioning in the isolated rat heart. AB - Transient glucose deprivation of the heart [GLU (-)] confers a preconditioning like protection against subsequent ischemic/reperfusion (I/R). The mechanisms involved remain unclear. We hypothesized that GLU (-) would induce the classic ischemic preconditioning activated signaling cascade. Potential metabolic consequences and putative cell signaling events induced by transient glucose deprivation were evaluated as candidate mediators of this cardioprotection. Isolated glucose-perfused rat hearts were subjected to 30 min global ischemia followed by 30 min reperfusion (index I/R). Cardiac contractile recovery following I/R was used as the functional end-point in these studies. Metabolic preconditioning was stimulated by 15 min GLU (-) followed by 10 min glucose repletion prior to the index I/R. The potential metabolic consequences of GLU (-) were evaluated by using excess octanoate (11 mM OCT Hi) or 11 mM 2-deoxy-D glucose (2-DG) in place of GLU (-) and by combining GLU (-) with fuels known to inhibit glycolysis supply (20 mM pyruvate or 1 mM octanoate, OCT Lo). The roles of alpha-adrenoceptors, beta-adrenoceptors, adenosine receptors, protein kinase C (PKC) and mitochondrial K(ATP) channels were investigated using inhibitors prazosin (10 microM), propranolol (10 microM), 8-(p-sulfophenyl) theophylline, (SPT 100 microM), chelerythrine (CHEL 10 microM) and 5-hydroxydecanoate (5 HD 100 microM) respectively. GLU (-) increased mechanical recovery (59.8 +/- 4.0 vs. 32.3 +/- 4.7%; p < 0.01). Protection was abolished by pyruvate 26.6 +/- 3.1; SPT 36.6 +/- 3.0; CHEL 35 +/- 4.8 or 5 HD 23.8 +/- 3.3%. In a separate set of experiments, the specificity of SPT in this model was tested by preconditioning with adenosine (100 microM) (34.7 +/- 4 vs. control 16.8 +/- 1.3%, p = 0.01) and blocking this protection with the same dose of SPT (16.3 +/- 1 .5%) used in the GLU (-) studies. Protection was unaltered by prazosin (50.2 +/- 3.3%), propranolol (55.5 +/- 4.0%), or OCT Lo (50.2 +/- 2.5%). Protection was not mimicked by OCT Hi (35.6 +/- 3.8%) or 2-DG (34 +/- 4.3%). Transient glucose deprivation does not seem to achieve preconditioning-like cardioprotection by decreased glycolysis. Rather, the signal system may involve enhanced adenosine release, PKC, and activation of the mitochondrial K(ATP) channel. PMID- 11055555 TI - Reciprocal regulation of glycogen phosphorylase and glycogen synthase by insulin involving phosphatidylinositol-3 kinase and protein phosphatase-1 in HepG2 cells. AB - The effect of insulin on glycogen synthesis and key enzymes of glycogen metabolism, glycogen phosphorylase and glycogen synthase, was studied in HepG2 cells. Insulin stimulated glycogen synthesis 1.83-3.30 fold depending on insulin concentration in the medium. Insulin caused a maximum of 65% decrease in glycogen phosphorylase 'a' and 110% increase in glycogen synthase activities in 5 min. Although significant changes in enzyme activities were observed with as low as 0.5 nM insulin level, the maximum effects were observed with 100 nM insulin. There was a significant inverse correlation between activities of glycogen phosphorylase 'a' and glycogen synthase 'a' (R2= 0.66, p < 0.001). Addition of 30 mM glucose caused a decrease in phosphorylase 'a' activity in the absence of insulin and this effect was additive with insulin up to 10 nM concentration. The inactivation of phosphorylase 'a' by insulin was prevented by wortmannin and rapamycin but not by PD98059. The activation of glycogen synthase by insulin was prevented by wortmannin but not by PD98059 or rapamycin. In fact, PD98059 slightly stimulated glycogen synthase activation by insulin. Under these experimental conditions, insulin decreased glycogen synthase kinase-3beta activity by 30-50% and activated more than 4-fold particulate protein phosphatase activity and 1.9-fold protein kinase B activity; changes in all of these enzyme activities were abolished by wortmannin. The inactivation of GSK-3beta and activation of PKB by insulin were associated with their phosphorylation and this was also reversed by wortmannin. The addition of protein phosphatase-1 inhibitors, okadaic acid and calyculin A, completely abolished the effects of insulin on both enzymes. These data suggest that stimulation of glycogen synthase by insulin in HepG2 cells is mediated through the PI-3 kinase pathway by activating PKB and PP-1G and inactivating GSK-3beta. On the other hand, inactivation of phosphorylase by insulin is mediated through the PI-3 kinase pathway involving a rapamycin-sensitive p70(s6k) and PP-1G. These experiments demonstrate that insulin regulates glycogen phosphorylase and glycogen synthase through (i) a common signaling pathway at least up to PI-3 kinase and bifurcates downstream and (ii) that PP-1 activity is essential for the effect of insulin. PMID- 11055556 TI - Protective effects of melatonin against oxidation of guanine bases in DNA and decreased microsomal membrane fluidity in rat liver induced by whole body ionizing radiation. AB - The aim of the study was to examine the potential protective effect of melatonin against whole body ionizing radiation (800 cGy). Changes in 8-hydroxy-2' deoxyguanosine (8-OH-dG) levels, an index of DNA damage, and alterations in membrane fluidity (the inverse of membrane rigidity) and lipid peroxidation in microsomal membranes, as indices of damage to lipid and protein molecules in membranes, were estimated. Measurements were made in rat liver, 12 h after their exposure to radiation. To test the potential protective effects of melatonin, the indole was injected (i.p. 50 mg/kg b.w.) at 120, 90, 60 and 30 min prior to radiation exposure. Both 8-OH-dG levels and microsomal membrane rigidity increased significantly 12 h after radiation exposure. Melatonin completely counteracted the effects of ionizing radiation. Changes in 8-OH-dG levels and membrane fluidity are early sensitive parameters of DNA and microsomal membrane damage, respectively, induced by ionizing radiation and our findings document the protective effects of melatonin against ionizing radiation. PMID- 11055557 TI - Evidence for the involvement of dietary lipids on the modulation of transforming growth factor-beta1 in the platelets of male rats. AB - Transforming growth factor beta1 (TGF-beta1), a multifunctional cytokine participates in the proliferation and differentiation of various cell types. Platelets are an important source of TGF-beta1 and are physiologically linked to a variety of chronic illnesses including cancer, heart disease and inflammation. It is well known that dietary lipids modulate platelet function. Whether dietary lipids affect growth factor status of platelets is not known. This study addresses the effect of dietary lipids on TGF-beta1 status of the platelets. Male 8 month-old Sprague Dawley rats were allocated to different diet groups. The high fat diets ( 18% by weight) comprising of high fat beef tallow (HFB), high fat corn oil (HFC), high fat fish oil (HFF) and high fat olive oil (HFO) and one low fat diet containing low fat soybean oil (LFS) (5% by weight) were fed to the experimental animals for 6 weeks. The TGF-beta1 status in the platelet lysate was assessed by using the CCL-64 mink lung cell bioassay and by Western blot analysis. Platelet lysates were evaluated for their ability to inhibit the growth of the CCL-64 mink lung cells, unexpectedly platelet lysates stimulated growth. The stimulatory effect of platelet lysate was in the order HFF > HFO > HFB > HFC > LFS. Acidification of the lysates to activate the latent form of TGF-beta1 resulted in the loss of the growth stimulatory potential of the platelet lysates in all the groups. Western blot analysis of the platelet lysates to detect the level of TGF-beta1 protein demonstrated that HFB diet group had the highest level of TGF-beta1 and the HFC diet group had the lowest level of TGF-beta1 and were significantly different (p < 0.05) as compared to the other three diet groups. These findings demonstrate that dietary lipids varying in their fatty acid composition, profoundly affect the level of growth modulating constituents of the platelets. Further studies are warranted to refine our understanding of the effect of dietary constituents on the physiology of the platelets. PMID- 11055558 TI - A call for snakebite research. PMID- 11055559 TI - The effect of capsaicin on the thermal and metabolic responses of men exposed to 38 degrees C for 120 minutes. AB - OBJECTIVE: To determine the thermoregulatory response of humans to heat following an ingestion of capsaicin. METHODS: The thermoregulatory responses of 7 men (aged 22-28 years) to a 2-hour exposure to 38 degrees C (50% relative humidity) were compared following ingestion of either placebo or capsaicin. The capsaicin dose (2 mg x kg(-1)) was ingested 1 hour prior to the heat exposure, and all subjects were encouraged to overhydrate for 48 hours prior to each trial. Core temperature, mean skin temperature, cardiac output, and oxygen consumption were measured every 30 minutes. In addition, the changes (pre-exposure vs postexposure) in body mass and plasma volume were calculated. RESULTS: As expected, core temperature, cardiac output, and oxygen consumption all increased significantly (P < .05) with exposure time. There were also significant decreases over time in body mass and plasma volume. These aforementioned changes, however, were not significantly different (P > .05) between the placebo and capsaicin trials, except for mean skin temperature. The treatment effect for mean skin temperature was significant (P = .013), with the capsaicin response (34.7 degrees C) registering lower than the placebo measurement (35.1 degrees C). CONCLUSIONS: It appears that humans respond differently than animals to capsaicin ingestion. For humans, ingesting a 2-mg x kg(-1) dose of capsaicin 1 hour prior to heat exposure does not alter a person's ability to thermoregulate in the heat. PMID- 11055560 TI - Thermal sensation and substrate utilization differs among low- and high-fat women exposed to 17 degrees C water. AB - OBJECTIVE: Thermal sensation and the physiological responses of women (follicular phase) exposed to 17 degrees C immersion for 120 minutes were investigated. METHODS: The subjects were divided into 2 groups by percent body fat (low fat [LF] = 21% +/- 2% [mean +/- SD] vs high fat [HF] = 30% +/- 3%). A 2-way analysis of variance was used to determine differences between the groups in metabolism, metabolism derived from carbohydrate, metabolism derived from fat, blood glucose, rectal temperature, skin temperature, and thermal sensation. RESULTS: As anticipated, pooled metabolism increased across the 120-minute immersion. Metabolism derived from carbohydrate was significantly higher in the LF than in the HF group and increased across time. Blood glucose decreased significantly across time, yet there was no group difference, suggesting that the LF group may have utilized a greater proportion of intramuscular glycogen. The HF group demonstrated a higher rectal temperature compared to their LF counterparts. Overall, rectal temperature demonstrated a group x time interaction as immersion continued. However, rectal temperature for all subjects remained above 35 degrees C. Surprisingly, the HF group perceived significantly greater thermal discomfort than did their LF counterparts. CONCLUSIONS: Since intramuscular glycogen utilization is associated with shivering thermogenesis, the suspected greater utilization of this fuel by the LF group may have contributed to less thermal discomfort than in the HF group. However, since glycogen utilization was not directly measured, this speculation cannot be validated. It is also possible that the modified thermal sensation scale we used may not be an adequate marker of thermal discomfort in females with a high percentage of body fat (28% to 35%) exposed to cold water immersion. PMID- 11055561 TI - Risk of immediate effects from F(ab)2 bivalent antivenin in Taiwan. AB - OBJECTIVE: To evaluate the incidence of immediate adverse effects from equine fragment antigen binding F(ab)2 bivalent antivenin produced by the National Institute of Preventive Medicine (NIPM) in Taiwan. METHODS: A retrospective chart review of patients presenting to a 600-bed general hospital over a 3-year period with snakebite who were treated with NIPM antivenin. RESULTS: A total of 130 snakebite victims presented to the emergency department over the study period, and 159 vials of antivenin were given. One hundred two patients (78.5%; 95% CI: 70, 85) received only hemorrhagic bivalent antivenin, 2 (1.5%; 95% CI: 0, 5) received only neurotoxic bivalent antivenin, and the remaining 26 (20.0%; 95% CI: 13, 28) received both kinds of bivalent antivenin. Three received a second vial of hemorrhagic antivenin because of progression of symptoms. Forty-two patients (32.3%; 95% CI: 24, 41) had positive skin tests, but following pretreatment with diphenhydramine and hydrocortisone, only 1 patient developed a skin rash thought to be related to antivenin. No patient developed an anaphylactic reaction. CONCLUSIONS: The use of NIPM F(ab)2 antivenin in snakebite victims in Taiwan has a very low risk of acute adverse reactions. PMID- 11055562 TI - Venomous snakebites in an urban area: what are the possibilities? AB - OBJECTIVE: To estimate the number of different species of venomous snakes privately kept in a large urban area. METHODS: An anonymous survey of potential snake owners in the Philadelphia urban and suburban area. The survey was mailed to members of the Philadelphia Herpetological Society. In addition, the survey was published in 2 herpetological newsletters and online in the Herpetology Network. RESULTS: One hundred sixty responses were obtained during a 6-month period. Ownership of 74 different varieties of venomous snakes was reported. Antivenin was not locally available for 13 of these species. CONCLUSION: A wide variety of venomous captive snakes can be found in the private sector. The potential for having to treat an envenomation requires the emergency physician to maintain an education of snakebite management options, including the various antivenin options available in their geographical location. PMID- 11055563 TI - Intermittent vs continuous hypoxia: effects on ventilation and erythropoiesis in humans. AB - OBJECTIVE: Recently, we showed that 5 days of normobaric intermittent hypoxia at rest (IH; 2 hours daily at 3,800 m simulated altitude; partial pressure of inspired oxygen 90 torr) can induce an increase in the isocapnic hypoxic ventilatory response (HVR) and blood reticulocyte count. The purpose of the present study was to compare these data with continuous exposure to the same hypoxic level. METHODS: Four of the same subjects were exposed, a year later, to 2 days of continuous hypoxia (CH), and 4 different subjects were exposed to 8 weeks of CH, both at the White Mountain Research Station (3,800 m altitude, barometric pressure approximately 489 torr). Inspired minute ventilation (VI), end-tidal partial pressure of carbon dioxide, arterial oxygen saturation (SaO2[sat]), hematocrit, and hemoglobin concentration were measured at different times during the continuous exposures. The HVR was expressed as the increase in V1 per 1% decrease in SaO2. RESULTS: The HVR showed no significant difference in the control values 1 year apart (IH, 0.06 +/- 0.03; CH2d (2 days' continuous hypoxia), 0.19 +/- 0.07 L x min(-1) x %sat(-1); means +/- SE), and the HVR values were similar after 2 days of IH compared to CH (0.42 +/- 0.26 and 0.51 +/- 0.22 L x min(-1) x %sat(-1), respectively). On the new subjects after 2 weeks of CH, the HVR showed a maximum increase, similar to the increase observed after only 5 days of IH, hemoglobin concentrations and hematocrit were significantly increased (45.0 +/- 2.7% vs 51.5 +/- 3.0% and 14.5 +/- 0.7 vs 17.2 +/- 1.0 g x dL(-1), respectively). The HVR did not change significantly from week 2 to 8 of CH, whereas hematological data were still increasing at the end of the 8 weeks. CONCLUSION: Changes in ventilatory oxygen sensitivity induced by IH and CH are similar in magnitude but occur with different time courses. The effects of IH on erythropoiesis are significant but fewer than on CH. PMID- 11055564 TI - Effects of a negative pressure venom extraction device (Extractor) on local tissue injury after artificial rattlesnake envenomation in a porcine model. AB - OBJECTIVES: To determine if a commercially available negative-pressure venom extraction device (Extractor) reduces local tissue injury after artificial rattlesnake envenomation in a porcine model. METHODS: We prospectively studied 10 pigs using a crossover design. After the pigs were anesthetized, 25 mg Crotalus atrox venom was injected obliquely with a 22-gauge needle 7 mm deep into subcutaneous tissues proximal to the ventral hind hoof. Pigs were randomized to receive either the Extractor (applied 3 minutes following envenomation and left in place for 30 minutes) or no Extractor. The protocol was repeated 14 days later by using the alternate treatment group and opposite hind leg for each animal. We measured leg circumference at standardized locations on the hoof, foreleg, and thigh at baseline and then 1, 2, 3, 4, 5, 6, 24, 48, 72, and 96 hours following venom injection. Maximal changes in circumference at 6 hours were compared using the paired t test. Minimum residual swelling at up to 96 hours was similarly compared. RESULTS: Maximal 6-hour swelling was similar with and without the Extractor: the hoof difference with the Extractor was -0.1% (95% CI = -3.4% to 3.2%, P = .95), foreleg difference was 0.3% (95% CI = -4.1% to 4.7%, P = .88), and thigh difference was -2.8% (95% CI = -10.0% to 4.4%, P = .40). Minimum residual swelling at up to 96 hours was also similar with and without the Extractor: hoof difference with the Extractor was 1.2% (95% CI = -5.6% to 8.0%, P = .70), foreleg difference was 0.6% (95% CI = -3.7% to 4.9%, P = .76), and thigh difference was 0.3% (95% CI = -2.4% to 3.0%, P = .81). A circular lesion identical in size and shape to the Extractor suction cup, which later necrosed and resulted in tissue loss, developed where the device had been applied in 2 animals. No such lesions occurred in legs not treated with the Extractor. CONCLUSION: No benefit was demonstrated from Extractor use for artificial rattlesnake envenomation in our animal study. The skin necrosis noted in 2 Extractor-treated extremities suggests that an injury pattern may be associated with the device. PMID- 11055565 TI - Oxygen concentrators for the delivery of supplemental oxygen in remote high altitude areas. AB - Oxygen concentrators are a relatively new technology for the delivery of supplemental oxygen. Readily available for domicile use in modern countries, these machines have proved reliable. The application of oxygen concentrators for the supply of medical oxygen in remote high-altitude settings has important cost saving and supply implications. In our experience at a remote hospital at 3,900 m in the Nepal Himalayas, oxygen concentrators constitute an effective and affordable means to supply medical oxygen. Using an air compressor and 2 zeolite chambers, the machine traps nitrogen from room air compressed to 4 atm, thus concentrating oxygen in the expressed gas. At delivery flow rates of 2 to 5 liters per minute, oxygen concentrations greater than 80% can be maintained. An electric power requirement of less than 400 W can be provided from a variety of sources, including a small gasoline generator, a solar or wind power system with battery store, or a domestic or commercial power source. At our facility, a cost savings of 75% for supplemental oxygen was found in favor of the oxygen concentrator over cylinders (0.17 US cents per liter vs 0.79 US cents per liter). PMID- 11055566 TI - Possible asphyxiation from carbon dioxide of a cross-country skier in eastern California: a deadly volcanic hazard. AB - This report describes an incident in which exceedingly high levels of carbon dioxide may have contributed to the death of a skier in eastern California. A cross-country skier was found dead inside a large, mostly covered snow cave, 1 day after he was reported missing. The autopsy report suggests that the skier died of acute pulmonary edema consistent with asphyxiation; carbon dioxide measurements inside the hole in which he was found reached 70%. This area is known for having a high carbon dioxide flux attributed to degassing of a large body of magma (molten rock) 10 to 20 km beneath the ski area. The literature describes many incidents of fatal carbon dioxide exposures associated with volcanic systems in other parts of the world. We believe this case represents the first reported death associated with volcanically produced carbon dioxide in the United States. Disaster and wilderness medicine specialists should be aware of and plan for this potential health hazard associated with active volcanoes. PMID- 11055567 TI - Myocardial infarction or high-altitude pulmonary edema? AB - We report the case of a 60-year-old European man with myocardial infarction at high altitude (4000 m). Myocardial infarction is an uncommonly encountered problem in high-altitude trekking in the Himalayas. The paucity of coronary artery disease at high altitude (hypoxia, exercise, and age not-withstanding) is discussed. Finally, the importance of recognizing disease entities that mimic acute mountain sickness in this environment is emphasized. PMID- 11055568 TI - Unusual presentation of a fracture and possible early compartment syndrome. PMID- 11055569 TI - A review of Buskirk's paper--a commentary. PMID- 11055570 TI - Work performance after dehydration: effects of physical conditioning and heat acclimatization. 1958. AB - Three groups of five men each were dehydrated overnight in the heat (115 degrees F) on two occasions (D1 and D2) to approximately 5.5% of their starting body weight. During the 3-week period between D1 and D2, one group (AC) was acclimatized to heat and physically conditioned, the second group (C) was physically conditioned and the third group (S) remained sedentary. The response to work after dehydration was assessed by the following criteria: pulse rate (P), rectal temperature (Tr) and maximal oxygen intake (Max. VO2). Pulse rates during and after walking and after running were elevated with dehydration. This elevation was reduced in groups AC and C at D2 as compared to D1, but not in group S. An elevation in T1 with walking also occurred with dehydration, but this elevation was not significantly different at D2 as compared with D1 in any group. Physical conditioning elicited an elevation in Max. VO2 (group AC and C), but the elevation was no greater in group AC than in group C. Dehydration was associated with an equal decrement in Max. VO2 at D1 and D2 in all groups, but the conditioned men (AC and C) maintained a relatively higher Max. VO2 than group S. Thus, physical conditioning was associated with enhanced work performance during dehydration (assessed by the above criteria), whereas acclimatization to heat did not appreciably supplement this effect. PMID- 11055571 TI - Teaching patient assessment. PMID- 11055572 TI - Severe hand injury following a green iguana bite. PMID- 11055573 TI - Janusz Supniewski as a researcher of psychotrophic drugs (on his 100th birthday). AB - On October 17, 1999, late Professor Janusz Supniewski's disciples celebrated the 100th anniversary of his birthday, and on June 3, 2000, they gathered to commemorate the 36th anniversary of his death. In our article we present one of the areas of Professor Supniewski's scientific activity, which is fruitfully pursued at the Institute of Pharmacology of the Polish Academy of Sciences, the successor of the Department of Pharmacology created by Supniewski in 1954. PMID- 11055574 TI - Memantine and ACPC affect conditioned place preference induced by cocaine in rats. AB - The influence of uncompetitive NMDA receptor antagonist, 1-amino-3,5--dimethyl adamantane (memantine) and partial glycineB site agonist, 1-amino cyclopropanecarboxylic acid (ACPC) on cocaine-induced conditioned place preference (CPP) were examined in male Wistar rats. After determination of initial preference, animals were conditioned with cocaine (5 mg/kg, ip) for 3 conditioning trials, alone or in combination with memantine (7.5 mg/kg, ip) or ACPC (50.0 mg/kg, ip). Memantine prevented acquisition and expression of the place preference produced by cocaine, while ACPC prevented only acquisition effect. Neither of the NMDA antagonists displayed any reinforcing properties by itself. Our current data suggest that the NMDA receptor complex may be involved in the rewarding effect of cocaine. PMID- 11055575 TI - Influence of opioid receptor antagonists on motor-impairing effects of benzodiazepines. AB - The influence of naloxone and naltrexone on the motor-impairing effects of diazepam, chlordiazepoxide, clonazepam and estazolam were studied in the aerial righting reflex test (mice, rats), and the first two drugs were examined in the rota-rod test (mice). Benzodiazepine-induced motor incoordination was significantly decreased by naloxone and naltrexone (4-16 mg/kg) in mice and rats in aerial righting reflex test. The motor-impairing effects of diazepam and chlordiazepoxide observed in rota-rod test were significantly diminished only by naltrexone (8-16 mg/kg). These data seem to confirm some interactions between benzodiazepines and opioid system. PMID- 11055576 TI - Trandolapril attenuates acquisition of conditioned avoidance in rats. AB - We have previously shown that angiotensin II (Ang II) improves cognitive performance of rats. In this study, behavioral effects of trandolapril, a highly lipophilic antihypertensive drug efficiently inhibiting brain angiotensin converting enzyme (ACE), were examined in rats. Single (0.1, 0.5, 1.0 mg/kg) or repetitive (0.1 mg/kg daily for 14 days) doses of trandolapril were given orally. The medium dose of the drug administered acutely as well as the chronic treatment significantly attenuated acquisition of conditioned avoidance responses. None of the three doses of trandolapril changed consolidation of memory and recall of the passive avoidance behavior, object recognition, and locomotor exploratory activity in open field. These data point to the psychoactive properties of trandolapril and suggest that physiological levels of the endogenous Ang II may be required for effective learning. PMID- 11055577 TI - Lack of effect of repeated treatment with a glycineB receptor partial agonist on the amphetamine-induced hyperactivity in rats. AB - The effect of acute and repeated (once daily, 14 days) administration of a potential antidepressant, the glycineB partial agonist 1 aminocyclopropanecarboxylic acid (ACPC, 100-400 mg/kg, ip), on the hyperactivity induced by amphetamine (0.5 mg/kg, sc) in rats was studied. Neither acute nor repeated treatment with the drug affected the hyperlocomotion induced by amphetamine. The obtained results indicate that ACPC does not resemble antidepressant drugs in this behavioral model. PMID- 11055578 TI - Influence of the aliphatic spacer length on the 5-HT1A receptor activity of new arylpiperazines with an indazole system. AB - Novel arylpiperazines (1a, 1c, 2a and 2c), containing a terminal 1- or 2 indazolyl fragment and a di- or tetramethylene aliphatic spacer, were synthesized and their 5-HT1A and 5-HT2A receptor affinities were determined. All those compounds showed a potent affinity for 5-HT1A receptors (Ki = 5-16 nM) and were evaluated for an intrinsic activity at those receptors. In order to determine a 5 HT1A agonistic effect of the investigated compounds, their ability to induce a lower lip retraction in rats and a behavioral syndrome (flat body posture and forepaw treading) in reserpinized rats were tested, whereas their 5-HT1A antagonistic activity was assessed via inhibition of those symptoms produced by 8 hydroxy-2-(di-n-propylamino)tetralin hydrobromide (8-OH-DPAT). The effect of spacer length on the 5-HT1A activity of the tested compounds was discussed in comparison with that of the three-methylene analogs (1b and 2b) described earlier. Both dimethylene derivatives (la and 2a) were characterized as weak postsynaptic 5-HT1A receptor antagonists. Compounds 1c (1-indazolyl analog) and 2c (2-indazolyl analog) with a tetramethylene aliphatic chain were classified as a postsynaptic 5-HT1A antagonist and a partial 5-HT1A agonist, respectively. Furthermore, the latter showed a moderate anxiolytic-like effect (conflict drinking Vogel's test in rats) and a weak antidepressant-like activity (forced swimming Porsolt's test in rats). PMID- 11055579 TI - Residual pulmonary neutrophils are involved in vaso- and bronchoconstrictor responses to platelet activating factor in the isolated buffer-perfused rat lung. AB - The aim of the study was to quantify residual neutrophils sequestrated in the isolated rat lung, perfused at a constant flow with Krebs-Hanseleit solution, and to analyze their possible influence on functional responses of the isolated lung. For that purpose we assessed neutrophil content in the isolated lung as well as in the effluent from the lung using an assay of myeloperoxidase (MPO) activity. We showed that a considerable pool of neutrophils remained in the isolated lung even after a 20-min period of washout with buffer solution. Moreover, these residual neutrophils were responsible for platelet activating factor (PAF) induced vaso- and bronchoconstriction but not for oedema formation. We conclude that when studying responses to pharmacological agents in isolated buffer perfused lung, the presence of sequestrated neutrophils should be taken into account. PMID- 11055580 TI - Effect of haloperidol treatment on immunoreactive neuropeptide Y secretion into lateral cerebral ventricle of rats. AB - The study aimed to evaluate the effect of acute and 14-day haloperidol treatment (2 mg/kg ip) on release of neuropeptide Y-like immunoreactivity (NPY-LI) in non anesthetized, freely moving rats by means of push-pull perfusion of the lateral cerebral ventricle. Twenty four hours after a single haloperidol injection NPY-LI release decreased by 15% (p < 0.05). No alterations were detected after 14-day haloperidol treatment. The study has confirmed that haloperidol alters the activity of neuropeptide Y system in the rat brain and suggested that acute haloperidol treatment inhibits the activity of the neuropeptide Y system at least in some brain structures surrounding the cerebral ventricles. The presented results have been discussed in view of our previous findings describing the haloperidol-induced changes in NPY-LI and NPY mRNA levels in the rat brain. PMID- 11055581 TI - Effect of repeated desipramine and fluoxetine administration on post-adjuvant arthritis. AB - The effects of desipramine and fluoxetine on the swelling of hind paws, radiologically-detectable bone destruction of hind paws, increase in spleen and popliteal lymph node weight, increase in metabolic activity of splenocytes and increase in proliferative activity of splenocytes and popliteal lymph node cells from right adjuvant injected paw in male C57BL/6 mice were studied on the 17th day after induction of post-adjuvant arthritis. Drugs were administered once daily ip at a dose of 10 mg/kg. Fourteen days of desipramine administration, starting on the third day after injection of the adjuvant, significantly increased edema and radiologically assessed bone destruction, spleen and popliteal lymph node weight whereas fluoxetine induced an opposite effect, but it did not reduce edema in comparison with saline-treated control. Two-week desipramine administration significantly increased metabolic activity of splenocytes and proliferative activity of popliteal lymph node cells from the right adjuvant-injected paw, whereas 14 days of fluoxetine injection reduced proliferative activity of splenocytes in comparison with the saline-treated mice. Desipramine administration 30 days before and 17 days after adjuvant injection did not change these parameters in spite of reduction of proliferative activity of splenocytes. These findings indicate that: 1) fluoxetine has a suppressive effect on some of the local and systemic changes which occur in adjuvant-induced arthritis in mice, 2) two-week desipramine administration significantly increases whereas 47-day desipramine treatment does not change most of local and systemic parameters of post-adjuvant disease in C57BL/6 mice, 3) the action of fluoxetine differs from that of desipramine in this model of autoimmunodisease probably as a result of the distinct effect of these two drugs on corticoids levels and on the activity of a sympathetic nervous system. PMID- 11055582 TI - Plasma levels of interleukin-6, interleukin-10, and interleukin-1 receptor antagonist in depression: comparison between the acute state and after remission. AB - There is now some evidence that major depression is accompanied by an immune response with an increased production of pro-inflammatory cytokines. The aim of the present study was to examine serum level of cytokines: interleukin-6 (IL-6), which is considered pro-inflammatory one and anti-inflammatory cytokines: interleukin 10 (IL-10), and interleukin-1 receptor antagonist (IL-1Ra) in acute clinical state of depression and after 6-week antidepressant treatment. Serum IL 6, IL-10, IL-1Ra levels were higher in the subjects with major depression than in normal controls although these results were not statistically significant. The mean score according to the Hamilton Depression Rating Scale (HDRS) in the patients significantly decreased during the 6 weeks of the study, demonstrating an overall improvement. Successful antidepressant treatment had no significant effect on serum level of this cytokines. PMID- 11055583 TI - Histone acetylation modifiers in the pathogenesis of malignant disease. AB - Chromatin structure is gaining increasing attention as a potential target in the treatment of cancer. Relaxation of the chromatin fiber facilitates transcription and is regulated by two competing enzymatic activities, histone acetyltransferases (HATs) and histone deacetylases (HDACs), which modify the acetylation state of histone proteins and other promoter-bound transcription factors. While HATs, which are frequently part of multisubunit coactivator complexes, lead to the relaxation of chromatin structure and transcriptional activation, HDACs tend to associate with multisubunit core-pressor complexes, which result in chromatin condensation and transcriptional repression of specific target genes. HATs and HDACs are known to be involved both in the pathogenesis as well as in the suppression of cancer. Some of the genes encoding these enzymes have been shown to be rearranged in the context of chromosomal translocations in human acute leukemias and solid tumors, where fusions of regulatory and coding regions of a variety of transcription factor genes result in completely new gene products that may interfere with regulatory cascades controlling cell growth and differentiation. On the other hand, some histone acetylation-modifying enzymes have been located within chromosomal regions that are particularly prone to chromosomal breaks. In these cases gains and losses of chromosomal material may affect the availability of functionally active HATs and HDACs, which in turn disturbs the tightly controlled equilibrium of histone acetylation. We review herein the recent achievements, which further help to elucidate the biological role of histone acetylation modifying enzymes and their potential impact on our current understanding of the molecular changes involved in the development of solid tumors and leukemias. PMID- 11055584 TI - Expression of the proteinase specialized in bone resorption, cathepsin K, in granulomatous inflammation. AB - BACKGROUND: The cysteine proteinase cathepsin K has aroused intense interest as the main effector in the digestion of extracellular matrix during bone resorption by osteoclasts. The enzyme is not a housekeeping lysosomal hydrolase, but is instead expressed with striking specificity in osteoclasts. In this work, we present evidence for the association of cathepsin K with the granulomatous reaction. Granulomas are inflammatory tissue reactions against persistent pathogens or foreign bodies. We came across cathepsin K while working on Echinococcus granulosus, a persistent tissue-dwelling, cyst-forming parasite that elicits a granulomatous response. MATERIALS AND METHODS: The walls of hydatid cysts from infected cattle were solubilized. Strong proteolytic activity was detected in the extracts. The proteinase responsible was purified by anion exchange and gel filtration. The purified protein was subjected to N-terminal sequencing, and its identity further confirmed by Western blotting, with a cathepsin K-specific antibody. The same antibody was used to localize the proteinase in paraffin-embedded sections of the parasite and the local host response. RESULTS: A proteinase was purified to near homogeneity from hydatid cyst extracts. The enzyme was unequivocally identified as host cathepsin K. Both the proenzyme and the mature enzyme forms were found. Cathepsin K was then immunolocalized both to the parasite cyst wall and to the epithelioid and giant multinucleated cells of the host granulomatous response. CONCLUSIONS: In the granulomatous response to the hydatid cyst, cathepsin K is expressed by epithelioid and giant multinucleated cells. We propose that, by analogy with bone resorption, cathepsin K is secreted by the host in an attempt to digest the persistent foreign body. Both processes, bone resorption and granulomatous reactions, therefore tackle persistent extracellular material (the bone matrix or the foreign body), and utilize specialized cells of the monocytic lineage (osteoclasts or epithelioid/giant cells) secreting cathepsin K as an effector. PMID- 11055585 TI - Chalcone, acyl hydrazide, and related amides kill cultured Trypanosoma brucei brucei. AB - BACKGROUND: Protozoan parasites of the genus Trypanosoma cause disease in a wide range of mammalian hosts. Trypanosoma brucei brucei, transmitted by tsetse fly to cattle, causes a disease (Nagana) of great economic importance in parts of Africa. T. b. brucei also serves as a model for related Trypanosoma species, which cause human sleeping sickness. MATERIALS AND METHODS: Chalcone and acyl hydrazide derivatives are known to retard the growth of Plasmodium falciparum in vitro and inhibit the malarial cysteine proteinase, falcipain. We tested the effects of these compounds on the growth of bloodstream forms of T. b. brucei in cell culture and in a murine trypanosomiasis model, and investigated their ability to inhibit trypanopain-Tb, the major cysteine proteinase of T. b. brucei. RESULTS: Several related chalcones, acyl hydrazides, and amides killed cultured bloodstream forms of T. b. brucei, with the most effective compound reducing parasite numbers by 50% relative to control populations at a concentration of 240 nM. The most effective inhibitors protected mice from an otherwise lethal T. b. brucei infection in an in vivo model of acute parasite infection. Many of the compounds also inhibited trypanopain-Tb, with the most effective inhibitor having a Ki value of 27 nM. Ki values for trypanopain-Tb inhibition were up to 50- to 100-fold lower than for inhibition of mammalian cathepsin L, suggesting the possibility of selective inhibition of the parasite enzyme. CONCLUSIONS: Chalcones, acyl hydrazides, and amides show promise as antitrypanosomal chemotherapeutic agents, with trypanopain-Tb possibly being one of their in vivo targets. PMID- 11055586 TI - Acute intermittent porphyria: expression of mutant and wild-type porphobilinogen deaminase in COS-1 cells. AB - BACKGROUND: Acute intermittent porphyria (AIP) is an autosomal dominant disorder that results from the partial deficiency of porphobilinogen deaminase (PBGD) in the heme biosynthetic pathway. Patients with AIP can experience acute attacks consisting of abdominal pain and various neuropsychiatric symptoms. Although molecular biological studies on the porphobilinogen deaminase (PBGD) gene have revealed several mutations responsible for AIP, the properties of mutant PBGD in eukaryotic expression systems have not been studied previously. MATERIALS AND METHODS: Seven mutations were analyzed using transient expression of the mutated polypeptides in COS-1 cells. The properties of mutated polypeptides were studied by enzyme activity measurement, Western blot analysis, pulse-chase experiments, and immunofluorescence staining. RESULTS: Of the mutants studied, R26C, R167W, R173W, R173Q, and R225X resulted in a decreased enzyme activity (0-5%), but R225G and 1073delA (elongated protein) displayed a significant residual activity of 16% and 50%, respectively. In Western blot analysis, the polyclonal PBGD antibody detected all mutant polypeptides except R225X, which was predicted to result in a truncated protein. In the pulse-chase experiment, the mutant polypeptides were as stable as the wild-type enzyme. In the immunofluorescence staining both wild-type and mutant polypeptides were diffusely dispersed in the cytoplasm and, thus, no accumulation of mutated proteins in the cellular compartments could be observed. CONCLUSIONS: The results confirm the causality of mutations for the half normal enzyme activity measured in the patients' erythrocytes. In contrast to the decreased enzyme activity, the majority of the mutations produced a detectable polypeptide, and the stability and the intracellular processing of the mutated polypeptides were both comparable to that of the wild-type PBGD and independent of the cross-reacting immunological material (CRIM) class. PMID- 11055587 TI - Insulin continues to induce plasminogen activator inhibitor 1 gene expression in insulin-resistant mice and adipocytes. AB - BACKGROUND: Although the association between insulin resistance and cardiovascular risk is well established, the underlying molecular mechanisms are poorly understood. The antifibrinolytic molecule plasminogen activator inhibitor 1 (PAI-1) is a cardiovascular risk factor that is consistently elevated in insulin-resistant states such as obesity and non-insulin-dependent diabetes mellitus (NIDDM). The strong positive correlation between this elevated PAI-1 and the degree of hyperinsulinemia not only implicates insulin itself in this increase, but also suggests that PAI-1 is regulated by a pathway that does not become insulin resistant. The data in this report supports this hypothesis. MATERIALS AND METHODS: We show that insulin stimulates PAI-1 gene expression in metabolically insulin-resistant ob/ob mice and in insulin-resistant 3T3-L1 adipocytes. Moreover, we provide evidence that glucose transport and PAI-1 gene expression are mediated by different insulin signaling pathways. These observations suggest that the compensatory hyperinsulinemia that is frequently associated with insulin-resistant states, directly contribute to the elevated PAI 1. CONCLUSIONS: These results provide a potential mechanism for the abnormal increases in cardiovascular risk genes in obesity, NIDDM, and polycystic ovary disease. PMID- 11055588 TI - A novel Ras antagonist regulates both oncogenic Ras and the tumor suppressor p53 in colon cancer cells. AB - BACKGROUND: In colon cancer, K-Ras oncogenes, which appear to be linked to chemoresistance and poor prognosis, are activated in more than 50% of cases, whereas the tumor suppressor gene p53 is mutationally altered in about 70% of all cases. The transcription factor p53, which is frequently mutated at codon 273, maintains wild-type configuration and possibly carries out residual functions. Although blocking of activated K-Ras may constitute a rational therapeutic concept for this treatment-resistant malignancy, a strategy influencing both oncogenic Ras and the tumor suppressor p53 may be even more promising. MATERIALS AND METHODS: We evaluated the effects of S-trans, trans-farnesyl-thiosalicylic acid (FTS), a novel Ras antagonist on human SW480 and HT-29 colon cancer cells, which both harbor a p53 His273 mutation but express activated K-Ras and wild type, but overexpressed, H-Ras, respectively. Besides cell growth and morphology, levels of cellular Ras proteins, regulation of p53 and p21(waf1/cip1) expression were analyzed by immunoblotting. The cell cycle arresting potential of FTS was quantified by flow cytometry. RESULTS: We demonstrate that FTS treatment alters the morphology and blocks the growth of SW480 and HT-29 colon cancer cells by both reducing the total amount of Ras and up-regulating the tumor suppressor p53. Furthermore, FTS caused an upregulation of the cyclin-cyclin-dependent kinase (CDK) inhibitor p21(waf1/cip1) and blocked the cell cycle. p53 antisense oligonucleotides not only reduced the level of p53 proteins but correspondingly also blocked the expression of p21(waf1/cip1) in FTS-treated colon cancer cells. CONCLUSIONS: FTS, a unique compound capable of regulating both oncogenic Ras and the tumor suppressor p53 may prove particularly useful for the therapy of colon cancer and other treatment-resistant malignancies where Ras is altered and p53 is either wild-type or mutated in positions that allow residual p53 functions. PMID- 11055589 TI - Suppression of type I collagen gene expression by prostaglandins in fibroblasts is mediated at the transcriptional level. AB - BACKGROUND: Tissues undergoing a chronic inflammatory process, such as the synovium in rheumatoid arthritis, are characterized by the infiltration of lymphocytes of different subsets and activation of monocyte/macrophages. Interleukin-1 (IL-1), a monocyte/ macrophage product that stimulates synovial fibroblasts to produce matrix metalloproteinases (MMPs), prostaglandins, and other cytokines, also has profound effects on the synthesis of extracellular matrix components such as type I collagen. In previous studies, we have shown that synovial fibroblasts and chondrocytes isolated from human joint tissues are particularly sensitive to prostaglandins, which modulate the effects of IL-1 on collagen gene expression in an autocrine manner. MATERIALS AND METHODS: BALBc/3T3 fibroblasts were treated with IL-1 and prostaglandins in the absence and presence of indomethacin to inhibit endogenous prostaglandin biosynthesis. Collagen synthesis was analyzed by SDS-PAGE as [3H]proline-labeled, secreted proteins, and prostaglandin production and cyclic adenosine 3',5'-cyclic monophosphate (camp) content were assayed. The expression of type I collagen gene (Col1a1) promoter reporter gene constructs was examined in transient transfection experiments, and the binding of nuclear factors to the Col1a1 promoter region spanning -222 bp/+ 116 bp was analyzed by DNase I footprinting and electrophoretic mobility shift (EMSA) assays. RESULTS: IL-1 increased the synthesis of type I and type III collagens in BALBc/3T3 fibroblasts; greater increases were observed when IL-1 stimulated synthesis of PGE2 was blocked by indomethacin. Transient transfection experiments demonstrated dose-dependent inhibition of the-222 bp Col1a1 promoter by exogenously added prostaglandins with the order of potency of PGF2alpha > PGE2 > PGE1 DNase I footprinting showed increased protection, which extended from the region immediately upstream of the TATA box, owing to the binding of nuclear factors from PGE2- or PGE1-treated BALBc/3T3 cells. EMSA analysis showed zinc dependent differences in the binding of nuclear factors from untreated and prostaglandin-treated cells to the -84 bp/-29 bp region of the Col1a1 promoter. CONCLUSIONS: These results show that the inhibition of Col1a1 expression by IL-1 in fibroblasts is mediated by prostaglandins at the transcriptional level and suggest that PGE-responsive factors may interact directly or indirectly with basal regulatory elements in the proximal promoter region of the Col1a1 gene. PMID- 11055590 TI - Business models in biotech. PMID- 11055591 TI - Competitive business intelligence gathering and analysis. PMID- 11055592 TI - Asthma. PMID- 11055593 TI - Xenotransplantation. PMID- 11055594 TI - A rebuttal: epidemic and endemic meningococcal meningitis in sub-Saharan Africa can be prevented now by routine immunization with group A meningococcal capsular polysaccharide vaccine. PMID- 11055595 TI - Yersinia enterocolitica infection in children. AB - BACKGROUND: Yersinia enterocolitica can cause illness ranging from self-limited enteritis to life-threatening systemic infection. The present study was undertaken to review the epidemiology, clinical manifestations, complications and outcome of Y. enterocolitica enteritis in children seen at a large children's hospital. METHODS: The project consisted of a retrospective chart review of medical and microbiologic records of all children with stool cultures positive for Y. enterocolitica during a 7-year period. RESULTS: The review included 142 patients with Y. enterocolitica enteritis. Patients' ages ranged from 18 days to 12 years, and the majority (85%) were younger than 1 year. Most patients presented during November, December and January. History of exposure to chitterlings (raw pork intestines) at home was elicited in 25 of 30 cases. Y. enterocolitica accounted for 12.6% (142 of 1,120) of all bacterial intestinal pathogens isolated during the study period. Blood cultures were positive in 7(9%) of 78 patients; 6 were younger than 1 year and one 12-year-old had sickle cell disease. Of 132 isolates tested all were susceptible to trimethoprim sulfamethoxazole, tobramycin and gentamicin; the majority were susceptible to cefotaxime (99%), ceftazidime (89%) and cefuroxime (88%). All bacteremic patients responded to cefotaxime treatment. Follow-up evaluation of 40 ambulatory patients revealed no difference in clinical improvement between those treated with oral trimethoprim-sulfamethoxazole (17 of 23) and those who were not treated (8 of 17) (P = 0.1). CONCLUSION: Y. enterocolitica is an important cause of enteritis in our young patient population during the winter holidays. Exposure of infants to chitterlings appears to be a risk factor. Infants younger than 3 months are at increased risk for bacteremia. Cefotaxime is effective in the treatment of Y. enterocolitica bacteremia; however, the role of oral antibiotics in the management of enteritis needs further evaluation. PMID- 11055596 TI - Pseudomonas aeruginosa bacteremia in children: analysis of trends in prevalence, antibiotic resistance and prognostic factors. AB - OBJECTIVE: To determine the factors predisposing to Pseudomonas aeruginosa bacteremia as well as the prevalence, source of infection, outcome and prognostic factors in pediatric patients. METHODS: Retrospective review of pediatric patients with P. aeruginosa bacteremia, at a large tertiary care hospital during a 6.5-year period. RESULTS: Seventy patients with P. aeruginosa bacteremia were identified. The annual rate of P. aeruginosa bacteremia remained unchanged during the study period. Antibiotic susceptibility remained unchanged except for two patients with extensive burns who developed resistant strains. Underlying diseases were malignancy (50%), prematurity (6%), burns (7%) and others (37%). The overall mortality associated with P. aeruginosa bacteremia was 20%. The fatality rate was higher among the young infants (compared with older children) and those who received previous antibiotic therapy (P = 0.02). Mortality rate was higher in nosocomial than in community-acquired infections (25% compared with 11.5%). The mortality rate of low birth weight and burns patients was significantly higher when compared with oncology patients or other patients, 75 and 40% compared with 11 and 19%, P = 0.01. Multiple regression analysis revealed a correlation only between the underlying disease and mortality (P = 0.02). In the oncology patients the only significant risk factor for mortality was absolute neutrophil count < or =0.1 x 10(9)/l (P = 0.06). CONCLUSION: P. aeruginosa bacteremia, although apparently not increasing in incidence and antibiotic resistance, is still a common serious complication in immunocompromised children with a high mortality rate. We conclude that the underlying disease is the main determinant of the clinical outcome. PMID- 11055597 TI - Short course therapy with cefitbuten versus azithromycin in pediatric streptococcal pharyngitis. AB - OBJECTIVE: To compare the safety and efficacy of a short course (5 days) of ceftibuten vs. azithromycin for 3 days for treatment of group A beta-hemolytic streptococcal (GABHS) pharyngitis in children. METHODS: A multicenter, open label, prospective, randomized trial in which patients > or =3 to < or =16 years of age with proven GABHS pharyngitis were randomized to receive either once daily ceftibuten for 5 days or azithromycin for 3 days. Patients were evaluated for clinical outcomes and/or for adverse events at days 6 to 8, 13 to 15 and 33 to 35 posttherapy. Microbiologic assessments (pharyngeal cultures) were conducted at baseline and at each follow-up visit. RESULTS: A total of 132 patients in the ceftibuten arm and 116 in the azithromycin arm were enrolled in the safety analysis, whereas 126 and 101, respectively, were enrolled for ceftibuten and azithromycin efficacy evaluation. Clinical success (cure or marked amelioration) at days 6 to 8 was recorded in 98 and 94% in the 2 groups, respectively. In the bacteriologic efficacy analysis at 6 to 8 days, the GABHS strain was eradicated in 76% of the patients treated with ceftibuten and in 76% of those receiving azithromycin. At 33 to 35 days, 84% of the patients in the ceftibuten arm and 71% in the azithromycin arm were GABHS-negative, and bacteriologic relapse was observed in 4 and 7% of the ceftibuten and azithromycin cases, respectively. Both treatments were well-tolerated by all patients. CONCLUSIONS: Ceftibuten and azithromycin allow simple treatment schedules (i.e. once daily administration, short duration of treatment). The somewhat higher eradication rate recorded after ceftibuten administration is consistent with the overall superior bactericidal activity of beta-lactams compared with macrolides vs. GABHS in vitro. PMID- 11055598 TI - Evaluation of Mycobacterium tuberculosis transmission from a pediatrician and initial compliance to prophylaxis of contacts in an outpatient pediatric clinic. AB - OBJECTIVE: The risk that latent infection will progress to active tuberculosis is greater in infants and children than for most other age groups. We set out to determine the rate of transmission of Mycobacterium tuberculosis to pediatric patients exposed to a pediatrician with smear-negative and culture-positive pulmonary tuberculosis. We also explored factors associated with compliance to prophylaxis. METHODS: Clinic and hospital billing records were used to identify patients age 5 or less who were seen during the pediatrician's potential contagious period. Patient were notified by registered mail of their putative exposure and were offered a tuberculin skin test screening with 5 tuberculin units of purified protein derivative (Tubersol, Connaught) and chest radiography of children with a tuberculin skin test > or =5 mm. RESULTS: A total of 456 patients were identified as exposed; 140 contacts never responded for evaluation and 93 letters were not delivered because of incorrect mailing addresses. Of the 223 who completed screening 1 (0,4%) had a initial skin test result of 8 mm. The remaining 222 contacts had repeated negative test results. The only positive child (15 months old) was born in Honduras and had received Calmette-Guerin bacillus at birth. No active tuberculosis cases were identified in the 456 contacts up to 2 years after exposure. Compliance with prophylaxis was associated with having two or less children in the household (odds ratio, 2.5; 95% confidence interval, 1.1 to 5.9). CONCLUSION: We found no evidence of transmission of M. tuberculosis in an outpatient pediatric setting. Only 43% of exposed children completed screening, and 38% of those offered prophylaxis completed their initial 3 months of therapy. PMID- 11055599 TI - Effect of human immunodeficiency virus infection on episodes of diarrhea among children in South Africa. AB - BACKGROUND: Infection with HIV is increasing among children in South Africa. Diarrhea is a common cause of morbidity and mortality in Africa, and some studies have shown that HIV-infected children have episodes of severe diarrhea with higher mortality than HIV-uninfected children. OBJECTIVES: To compare the severity, pathogens and outcome of diarrhea in HIV-infected and uninfected children. METHODS: We studied 181 children ages 3 months to 4 years admitted for gastroenteritis to the Chris Hani Baragwanath Hospital in Soweto, South Africa. Demographic details of the children were recorded, as were the details of the episode of diarrhea. Stools specimens were collected and sent for microbiologic evaluation. The clinical course of the child's admission was recorded. Children were diagnosed as being infected with HIV if they tested positive by HIV enzyme linked immunosorbent assay (ELISA) and were >15 months of age, or if they were ELISA-positive, were < 15 months of age and had clinical signs of HIV infection. RESULTS: Of the 176 children with an HIV ELISA result, 31 (17.6%) were classified as HIV-infected. More HIV-infected children were malnourished (80.6% vs. 39.5%, P < 0.001) and more likely to have had prolonged diarrhea (16.1% vs. 5.9%, P = 0.07) compared with HIV-uninfected children. HIV-infected children had a higher rate of a codiagnosis of pneumonia (43.3% vs. 9.2%, P < 0.0001) and were more likely to require a hospital stay of >4 days (prevalence odds ratio, 5.11; 95% confidence interval, CI 1.49 to 17.52). There were no significant differences in stool pathogens or in the level of dehydration on admission between the HIV infected and uninfected children. CONCLUSION: HIV-infected children have the same spectrum of enteric pathogens as uninfected children but require more attention because of malnutrition and comorbidity. PMID- 11055600 TI - Cross-sectional survey of trachoma in school age children in the region of Thies (Senegal). AB - BACKGROUND: Trachoma is a leading cause of blindness in West Africa. In Senegal previous studies have shown that the endemicity is high. AIMS: To verify the extent of the pathology and to study the epidemiologic characters of this infectious disease in a population of school age children in a rural zone, located in the Thies region (Senegal). METHODS: A cross-sectional survey in six villages in the region of Thies was performed in a population of school age children (5 to 15 years old). Sixty variables (individual, family, village, etc.) with morbidity indicators for trachoma cases were collected and analyzed. RESULTS: The cross-sectional survey confirmed the extent of the prevalence of trachoma in the childhood population; 208 of the 388 children (mean age, 9.19 years) included had trachoma (53.6%). Significant statistical correlations were found among the occurrence of trachoma and ethnic origins, the village of residence, the father's profession and the daily quantity of water ingested and its origin. Inversely no correlation could be found between the occurrence of trachoma and sex, age (except for florid trachoma), the size of the sibship, whether the mother washes the child, washing their hands before eating, the number of cospouses and the number of children per mother. CONCLUSIONS: The data obtained confirmed the extent of this endemic disease in the region of Thies, because dispensing antibiotic eye drop has limited efficacy over time. Information should be provided to the population on the extent of the disease and its epidemiologic characteristics and more widespread well drilling, and use of that water should be encouraged. PMID- 11055601 TI - Household crowding a major risk factor for epidemic meningococcal disease in Auckland children. AB - BACKGROUND: New Zealand is in its ninth year of a serogroup B meningococcal disease epidemic with annual rates of up to 16.9 cases per 100,000. The highest incidence is in Maori and Pacific Island children in the Auckland region. We conducted a case-control study to identify potentially modifiable risk factors for this disease. METHODS: A case-control study of 202 cases of confirmed and probable meningococcal disease in Auckland children younger than 8 years of age recruited from May, 1997, to March, 1999, was undertaken. Controls (313) were recruited door-to-door by a cluster sampling method based on starting points randomly distributed in the Auckland region. They were frequency matched with the expected distribution of age and ethnicity in the meningococcal disease cases. RESULTS: With the use of a multivariate model and controlling for age, ethnicity, season and socioeconomic factors, risk of disease was strongly associated with overcrowding as measured by the number of adolescent and adult (10 years or older) household members per room [odds ratio (OR), 10.7; 95% confidence interval (CI), 3.9 to 29.5]. This would result in a doubling of risk with the addition of 2 adolescents or adults to a 6-room house. Risk of disease was also associated with analgesic use by the child, which was thought to be a marker of recent illness (OR 2.4, CI 1.5 to 4.0); number of days at substantial social gatherings (10 or more people for > 4 h; OR 1.8, CI 1.2 to 2.6); number of smokers in the household (OR 1.4, CI 1.0 to 1.8); sharing an item of food, drink or a pacifier (OR 1.6, CI 1.0 to 2.7); and preceding symptoms of a respiratory infection (cough, "cold or flu," runny nose, sneezing) in a household member (OR 1.5, CI 1.0 to 2.5). CONCLUSION: Some of these identified risk factors for meningococcal disease are modifiable. Measures to reduce overcrowding could have a marked effect on reducing the incidence of this disease in Auckland children. PMID- 11055602 TI - Predictors of infectious complications after burn injuries in children. AB - BACKGROUND: Infections are the major life-threatening complication of burn injury and occur with the greatest frequency in children. Knowledge of their occurrence and management, however, is extrapolated from studies in adults. We performed a prospective study of infectious complications in burned children. OBJECTIVE: To delineate epidemiology, risk factors and microbiology of infections in burned children where burn care and surgical interventions are optimal. METHODS: Children hospitalized for burns were entered into prospective study. Characteristics of the burn injury were assessed, and active surveillance for infections was performed. RESULTS: Seventy patients were entered [mean age, 42 months; mean total body surface area (TBSA), burn 15%]. Twenty-seven percent of patients developed 39 infections: 13 involved the burn wound (burn wound sepsis, 6; graft loss, 5; and cellulitis, 2); 13 were catheter-associated septicemia; 13 involved other sites (i.e. pneumonia, 4; urinary tract infection, 3; bacteremia, 2; endocarditis, 1; myocardial abscess, 1; toxin-mediated syndrome, 1; and otitis media, 1). Twenty-three infections were caused by a single organism, 9 infections by more than 1 organism and in 7 infections defined by CDC criteria no organism was recovered. Organisms causing infection were: Staphylococcus aureus, 19; Candida albicans, 4; Pseudomonas aeruginosa, 4; coagulase-negative Staphylococcus, 4; Enterococcus sp., 3; Escherichia coli, 1; Klebsiella oxytoca, 1; Serratia marcescens, 1; Streptococcus pneumoniae, 1; Streptococcus pyogenes, 1; Aspergillus fumigatus, 1; and Candida parapsilosis, 1. Burn mechanism (flame and inhalation), extent (TBSA >30%) and depth (full thickness) were risk factors for infection; young age and site of burn were not. CONCLUSION: The most common infections occurring in burn children are burn wound infections and catheter associated septicemia. Characteristics of burn injury predict risk of infection. Children with flame and inhalation injury, TBSA burned >30% and full thickness burns are at high risk of infectious complications. PMID- 11055603 TI - Adenovirus 7a: a community-acquired outbreak in a children's hospital. AB - BACKGROUND: Adenoviruses produce many illnesses in children, particularly respiratory and gastrointestinal disease. The most common adenoviral respiratory infections in children are caused by types 1, 2, 3 and 5. Adenoviruses spread rapidly in closed environments often causing epidemic disease. Serotype 7a has been responsible for outbreaks of respiratory disease in children living in close proximity with one another. This report describes a large community-acquired adenovirus 7a epidemic in hospitalized children. METHODS: Evaluation of all patients with cultures positive for adenovirus from a children's hospital-based virology laboratory during a recognized adenovirus outbreak. All such adenovirus isolates were typed, and patients with adenovirus 7a are described by review of medical records. RESULTS: Between March 1 and July 26, 1997, 47 children admitted to the hospital were identified as infected with adenovirus. Of these 47 patients 26 (55%) were infected with adenovirus 7a. Twenty-four (92%) infections were community-acquired. The age range was 11 days to 10 years with a median of 9.5 months. Twenty-two patients (84%) had respiratory symptoms, and 21 (8%) had fever, making these the most common symptoms. The mean durations of fever and hospitalization were 5.5 and 7 days, respectively. One of 26 patients died. CONCLUSIONS: Adenovirus 7a can cause large community epidemics affecting children. The disease produced by adenovirus 7a in children is almost exclusively of the respiratory tract, and in some individuals it may be very severe and possibly fatal. PMID- 11055604 TI - Risk factors for sternal wound and other infections in pediatric cardiac surgery patients. AB - BACKGROUND: This study was undertaken to determine the incidence, pathogens and risk factors associated with development of sternal wound and other infections in children undergoing cardiac surgery. METHODS: Retrospective chart review was performed for all cardiac surgeries performed on children <18 years of age at Upstate Medical University at Syracuse between January, 1996, and June, 1998. For evaluation of risk factors for sternal wound infection, only patients undergoing sternotomy are included in the analysis: those with infection are compared with those without for preoperative, intraoperative and postoperative risk factors. RESULTS: Sternal wound infection developed in 10 of 202 (5%) children after median sternotomy. Superficial sternal wound infection developed in 6 (3%) children, and 4 (2%) had deep infection. Children with sternal wound infection had lower age, higher American Society of Anesthesiologist score, longer preoperative stay, longer period of ventilation and inotropic support, longer intensive care unit and total postoperative hospital stays and increased leukocyte band cell counts preoperatively and on Postoperative Day 1 than those without sternal infection. Causative agents for sternal wound infection were Staphylococcus aureus (6), Pseudomonas aeruginosa (1) and Haemophilus influenzae non-type b (1). In addition 32 bacterial infections occurred at nonsurgical sites after 28 procedures. Infections included pneumonia, urinary tract infection and bacteremia. Longer bypass time and longer operation time were two additional risk factors for nonwound infection. CONCLUSION: Infections continue to be a significant cause of morbidity in cardiac surgery patients. Knowledge of risk factors for infection could be useful in preventive and treatment strategies for these high-risk groups. PMID- 11055605 TI - Viruses and acute otitis media. PMID- 11055606 TI - Management of otorrhea in infants and children. PMID- 11055607 TI - Pyomyositis in children. PMID- 11055608 TI - Diagnosis of human immunodeficiency virus infection in perinatally exposed orphaned infants in a resource-poor setting. PMID- 11055609 TI - Bacteriologic outcome of children with cefotaxime- or ceftriaxone-susceptible and -nonsusceptible Streptococcus pneumoniae meningitis. PMID- 11055610 TI - Lack of utility of serotyping multiple colonies for detection of simultaneous nasopharyngeal carriage of different pneumococcal serotypes. PMID- 11055611 TI - Incidence of invasive Haemophilus influenzae type b infections in Filipino children. PMID- 11055612 TI - Sepsis after nasolarcrimal duct probing. PMID- 11055613 TI - Lymphocutaneous Nocardia brasiliensis infection: a pediatric case cured with amoxicilin/clavulanate. PMID- 11055614 TI - Trichosporon inkin lung abscesses presenting as a penetrating chest wall mass. PMID- 11055615 TI - Disseminated fungal infection associated with myeloperoxidase deficiency in a premature neonate. PMID- 11055616 TI - Postexposure varicella vaccination in nine-month-old children. PMID- 11055617 TI - Severity of respiratory syncytial virus disease. PMID- 11055618 TI - Treatment of otitis media. PMID- 11055619 TI - Should your neighbor buy a gun? PMID- 11055620 TI - Are schizophrenia or antipsychotic drugs a risk factor for cataracts? AB - Lens changes and ocular disturbances have been reported in conjunction with the use of antipsychotic drugs. We estimated the incidence rate of a clinical diagnosis of cataract in patients with a psychotic disorder, schizophrenia, and compared it with the rate in the general population. Among the schizophrenic patients, we also examined the role of dose and duration of antipsychotic drugs on the risk of cataract development. We followed up two cohorts of patients 30-85 years of age who were included in the United Kingdom General Practice Research Database. Patients in one group had a diagnosis of schizophrenia (N = 4,209). The other group was an age- and sex-matched cohort of 10,000 patients sampled from the source population. The incidence of cataracts was 4.5 per 1,000 person-years among the general population and 3.5 in the schizophrenia population. Overall, antipsychotic drug use was not associated with the occurrence of cataracts. Nevertheless, among long-term users of chlorpromazine at daily doses of 300 mg or greater, and among users of prochlorperazine, the relative risks were 8.8 (95% confidence interval = 3.1-25.1) and 4.0 (95% confidence interval = 0.8-20.7), respectively. There is no indication that schizophrenia per se is associated with an increased risk of developing cataracts. PMID- 11055621 TI - A pooled analysis of magnetic fields, wire codes, and childhood leukemia. Childhood Leukemia-EMF Study Group. AB - We obtained original individual data from 15 studies of magnetic fields or wire codes and childhood leukemia, and we estimated magnetic field exposure for subjects with sufficient data to do so. Summary estimates from 12 studies that supplied magnetic field measures exhibited little or no association of magnetic fields with leukemia when comparing 0.1-0.2 and 0.2-0.3 microtesla (microT) categories with the 0-0.1 microT category, but the Mantel-Haenszel summary odds ratio comparing >0.3 microT to 0-0.1 microT was 1.7 (95% confidence limits = 1.2, 2.3). Similar results were obtained using covariate adjustment and spline regression. The study-specific relations appeared consistent despite the numerous methodologic differences among the studies. The association of wire codes with leukemia varied considerably across studies, with odds ratio estimates for very high current vs low current configurations ranging from 0.7 to 3.0 (homogeneity P = 0.005). Based on a survey of household magnetic fields, an estimate of the U.S. population attributable fraction of childhood leukemia associated with residential exposure is 3% (95% confidence limits = -2%, 8%). Our results contradict the idea that the magnetic field association with leukemia is less consistent than the wire code association with leukemia, although analysis of the four studies with both measures indicates that the wire code association is not explained by measured fields. The results also suggest that appreciable magnetic field effects, if any, may be concentrated among relatively high and uncommon exposures, and that studies of highly exposed populations would be needed to clarify the relation of magnetic fields to childhood leukemia. PMID- 11055622 TI - Estrogen metabolism and risk of breast cancer: a prospective study of the 2:16alpha-hydroxyestrone ratio in premenopausal and postmenopausal women. AB - Experimental and clinical evidence suggests that 16alpha-hydroxylated estrogen metabolites, biologically strong estrogens, are associated with breast cancer risk, while 2-hydroxylated metabolites, with lower estrogenic activity, are weakly related to this disease. This study analyzes the association of breast cancer risk with estrogen metabolism, expressed as the ratio of 2-hydroxyestrone to 16alpha-hydroxyestrone, in a prospective nested case-control study. Between 1987 and 1992, 10,786 women (ages 35-69 years) were recruited to a prospective study on breast cancer in Italy, the "Hormones and Diet in the Etiology of Breast Cancer" (ORDET) study. Women with a history of cancer and women on hormone therapy were excluded at baseline. At recruitment, overnight urine was collected from all participants and stored at -80 degrees C. After an average of 5.5 years of follow-up, 144 breast cancer cases and four matched controls for each case were identified among the participants of the cohort. Among premenopausal women, a higher ratio of 2-hydroxyestrone to 16alpha-hydroxyestrone at baseline was associated with a reduced risk of breast cancer: women in the highest quintile of the ratio had an adjusted odds ratio (OR) for breast cancer of 0.58 [95% confidence interval (CI) = 0.25-1.34]. The corresponding adjusted OR in postmenopausal women was 1.29 (95% CI = 0.53-3.10). Results of this prospective study support the hypothesis that the estrogen metabolism pathway favoring 2 hydroxylation over 16alpha-hydroxylation is associated with a reduced risk of invasive breast cancer risk in premenopausal women. PMID- 11055623 TI - Age at menarche and tanner stage in girls exposed in utero and postnatally to polybrominated biphenyl. AB - Accidental contamination of the Michigan food chain with polybrominated biphenyls (PBBs) led to the exposure of more than 4,000 individuals in 1973. Because PBB exposure is suspected to disrupt endocrine function, we assessed pubertal development in females 5-24 years of age (N = 327) who were exposed to PBB in utero and, in many cases, through breastfeeding. We estimated in utero PBB exposure using maternal serum PBB measurements taken after exposure (1976-1979) and extrapolated to time of pregnancy using a model of PBB decay. We found that breastfed girls exposed to high levels of PBB in utero (> or =7 parts per billion) had an earlier age at menarche (mean age = 11.6 years) than breastfed girls exposed to lower levels of PBB in utero (mean age = 12.2-12.6 years) or girls who were not breastfed (mean age = 12.7 years). This association persisted after adjustment for potential confounders (menarche ratio = 3.4, 95% confidence interval = 1.3-9.0). Perinatal PBB exposure was associated with earlier pubic hair stage in breastfed girls, but little association was found with breast development. The associations observed here lend support to the hypothesis that pubertal events may be affected by pre- and postnatal exposure to organohalogens. PMID- 11055624 TI - Metabolic gene polymorphisms and risk of dysmenorrhea. AB - We conducted a molecular epidemiologic study in rural China to investigate the association of the cytochrome P450 2D6 (CYP2D6) and glutathione S-transferase Mu (GSTM1) polymorphisms with dysmenorrhea. This report includes 435 subjects, 129 with and 306 without any history of dysmenorrhea, who did not smoke or drink alcohol. We obtained information on dysmenorrhea and major covariates by questionnaire interview. We used categorical methods and logistic regression models to evaluate the individual and combined associations of CYP2D6 and GSTM1 polymorphisms with dysmenorrhea and its subgroups, occasional (N = 70) and recurrent (N = 59), with adjustment for age, education, occupation, passive smoke exposure, age of menarche, parity, contraceptive method, height, and body mass index. Both variant CYP2D6 and GSTM1 genotypes were associated with increased risk of recurrent dysmenorrhea [for CYP2D6, odds ratio (OR) = 1.7 and 95% confidence interval (95% CI) = 0.9-3.1; for GSTM1, OR = 1.8 and 95% CI = 1.0 3.4). There was no appreciable association between these variant genotypes and occasional dysmenorrhea. When both the CYP2D6 and GSTM1 genotypes were considered together, the highest risk of recurrent dysmenorrhea was found among women with variant genotypes in both CYP2D6 and GSTM1 (OR = 3.1; 95% CI = 1.2-8.0). This study provides evidence of genetic susceptibility to recurrent dysmenorrhea. PMID- 11055625 TI - Recommendations for the design of epidemiologic studies of endometriosis. AB - This paper proposes a standard definition of endometriotic disease for epidemiologic studies and suggests subject-selection strategies to increase the validity of clinic- or population-based studies of the disease. Although endometriosis can be defined simply as the presence of ectopic endometrial tissue, emerging evidence indicates that to be pathologic, such tissue must persist and progress. The proposed disease definition incorporates the concepts of persistence and progression, and its use may increase the likelihood of observing true associations in etiologic studies. Potential threats to validity of substantial magnitude exist in both clinic- and population-based epidemiologic studies of endometriosis. In clinic-based studies, control subjects (infertility clinic patients, women delivering infants, or women undergoing tubal ligation) often are not representative of the population from which the cases arose, and bias can be considerable for behavioral and hormone-related exposures. In population-based studies, substantial case underascertainment may exist, and diagnosed cases may be a biased sample of all potential cases in the population. Although neither the ideal design nor the ideal case and control groups are likely to be achievable in epidemiologic studies of endometriosis, the subject selection strategies suggested may improve the validity of studies that are obliged to depart from the ideal. PMID- 11055626 TI - Alcohol consumption and urinary concentration of 6-sulfatoxymelatonin in healthy women. AB - Consumption of alcoholic beverages may suppress circulating melatonin levels at night, possibly resulting in an increase in circulating estrogen. An increased estrogen burden could increase the risk of breast cancer. This study was designed to investigate whether alcohol consumption is associated with a decrease in nighttime melatonin levels in a group of healthy women. A total of 203 randomly selected healthy women between the ages of 20 and 74 years were recruited for a broader study of the effects of exposure to power-frequency magnetic fields on nocturnal levels of urinary 6-sulfatoxymelatonin. For the purposes of this analysis, data collection consisted of the following during two seasons of the year: (1) an in-person interview, (2) a daily activity diary, and (3) nocturnal urine collection for each of 3 consecutive nights. We found that the nocturnal urinary concentration of the primary metabolite of melatonin (6 sulfatoxymelatonin) decreased in a dose-dependent manner with increasing consumption of alcoholic beverages in the preceding 24-hour period, after taking into account the independent effects on melatonin of age, hours of darkness, use of medications that affect melatonin levels, and body mass index. A categorical analysis revealed no effect of one drink, but a 9% reduction with two drinks, a 15% reduction with three drinks, and a 17% reduction with four or more drinks. It remains unknown whether such a change could affect estrogen levels or breast cancer risk. PMID- 11055627 TI - Using meta-smoothing to estimate dose-response trends across multiple studies, with application to air pollution and daily death. AB - Air pollution has been associated with daily mortality in numerous studies over the last decade. Although considerable attention has focused on issues of potential confounding in these associations, little has been done to address the question of what the shape of the dose-response relation looks like. The question of whether a threshold exists for these relations is of particular concern, with regard to both this application and many other epidemiologic questions. Nonparametric smoothing is widely used to control for the potentially nonlinear relations between covariates and daily deaths but has been little used to model the air pollution associations. Because sampling variability, among other factors, can introduce considerable noise into the estimates of linear dose response curves, quantitative summaries have been widely used to come up with best linear fits. The same ability of meta-analytic techniques to average out noise applies to nonparametric smooth estimates in individual cities. We have developed a method of applying these techniques to combining nonparametric smooths. Using simulation studies, we show that this method can detect threshold and other nonlinear relations in epidemiologic studies, and we then apply it to analyze the association between PM10 and daily deaths in ten U.S. cities. We find that the association appears linear down to the lowest levels observed in the study. This method is generally applicable in settings where data from multiple studies can be combined. PMID- 11055628 TI - Lung cancer and arsenic concentrations in drinking water in Chile. AB - Cities in northern Chile had arsenic concentrations of 860 microg/liter in drinking water in the period 1958-1970. Concentrations have since been reduced to 40 microg/liter. We investigated the relation between lung cancer and arsenic in drinking water in northern Chile in a case-control study involving patients diagnosed with lung cancer between 1994 and 1996 and frequency-matched hospital controls. The study identified 152 lung cancer cases and 419 controls. Participants were interviewed regarding drinking water sources, cigarette smoking, and other variables. Logistic regression analysis revealed a clear trend in lung cancer odds ratios and 95% confidence intervals (CIs) with increasing concentration of arsenic in drinking water, as follows: 1, 1.6 (95% CI = 0.5 5.3), 3.9 (95% CI = 1.2-12.3), 5.2 (95% CI = 2.3-11.7), and 8.9 (95% CI = 4.0 19.6), for arsenic concentrations ranging from less than 10 microg/liter to a 65 year average concentration of 200-400 microg/liter. There was evidence of synergy between cigarette smoking and ingestion of arsenic in drinking water; the odds ratio for lung cancer was 32.0 (95% CI = 7.2-198.0) among smokers exposed to more than 200 microg/liter of arsenic in drinking water (lifetime average) compared with nonsmokers exposed to less than 50 microg/liter. This study provides strong evidence that ingestion of inorganic arsenic is associated with human lung cancer. PMID- 11055629 TI - Birth order, as a proxy for age at infection, in the etiology of hepatocellular carcinoma. AB - First-born and second-born children are exposed to common infections after enrollment at school, whereas later-born children are exposed to these infections earlier through their older siblings. We have evaluated whether birth order is a risk factor for hepatitis B virus (HBV)-related, hepatitis C virus (HCV)-related, and apparently virus-unrelated hepatocellular carcinoma (HCC) in a large case control study that included 333 HCC cases and 632 controls. In comparison with controls who were carriers of hepatitis B surface antigen (HBsAg), HBsAg-positive HCC cases were more likely to have been later-born children (odds ratio per increase in birth order = 2.0; 95% confidence interval = 1.2-3.6). There was no such evidence for anti-HCV-positive cases compared with anti-HCV-positive controls or for virus-negative HCC cases compared with virus-negative controls. We conclude that early infection with HBV increases the risk of HBV carriers to develop HCC, over and beyond its role in facilitating the establishment of a carrier state. PMID- 11055630 TI - Multilevel modeling in epidemiology with GLIMMIX. AB - Previous work has shown that multilevel modeling can be a valuable technique for epidemiologic analysis. The complexity of using this approach, however, continues to restrict its general application. A critical factor is the lack of flexible and appropriate software for multilevel modeling. SAS provides a macro, GLIMMIX, that can be used for multilevel modeling, but that is not sufficient for a complete epidemiologic analysis. We here provide additional code to obtain epidemiologic output from GLIMMIX, illustrated with new data on diet and breast cancer from the European Community Multicenter Study on Antioxidants, Myocardial Infarction, and Breast Cancer (EURAMIC). Our results give epidemiologists an easily used tool for fitting multilevel models. PMID- 11055631 TI - A prospective study of the risk of congenital defects associated with maternal obesity and diabetes mellitus. AB - This study was designed to evaluate the effects of maternal obesity and diabetes mellitus on the risk of nonchromosomal congenital defects. We used data from 22,951 pregnant women enrolled in a prospective cohort study of early prenatal exposures and pregnancy outcome. The relative risks [prevalence ratios (PRs)] of major nonchromosomal congenital defects associated with obesity and diabetes, alone or in combination, were calculated using multiple logistic regression analysis. In this study, in the absence of diabetes, obese women (body mass index > or =28) had no higher risk, overall, of having an offspring with a major defect [PR = 0.95; 95% confidence interval (CI) = 0.62-1.5]. Their offspring, however, did have a higher prevalence of certain types of defects, including orofacial clefts; club foot; cardiac septal defects; and, to a lesser extent, hydrocephaly and abdominal wall defects. Women with pre-existing or gestational diabetes who were not obese also had no excess risk overall of having offspring affected by a major defect (PR = 0.98; 95% CI = 0.43-2.2), although they did have a higher prevalence of musculoskeletal defects. The pregnancies of women who were both obese and diabetic were 3.1 times as likely (95% CI = 1.2-7.6) to result in an offspring with a defect than were those of nonobese, nondiabetic women, which suggests that obesity and diabetes mellitus may act synergistically in the pathogenesis of congenital anomalies. The defects were largely craniofacial or musculoskeletal. PMID- 11055632 TI - Effect of prenatal diagnosis on epidemiologic studies of birth defects. AB - Prenatal diagnostic technology makes it possible to offer women the option of electively terminating pregnancies affected by birth defects. Excluding these pregnancies from epidemiologic studies may affect study results. We explored this effect using examples from the literature. We calculated the bias in the odds ratio caused by excluding prenatally diagnosed pregnancies when the exposure of interest is not correlated with the likelihood of terminating an affected pregnancy and when it is correlated with an increase or decrease in this likelihood. We assumed that control infants did not have birth defects. When the exposure is not associated with the likelihood of a pregnancy termination, studies excluding terminations suffer a loss of precision. When the exposure is associated with an increase or decrease in this likelihood, the odds ratios are biased toward or away from the null, respectively. The magnitude of the bias will vary according to characteristics of the study population such as the prevalence of the exposure and the frequency with which prenatal diagnosis and elective termination are used. Whenever possible, pregnancies terminated after prenatal diagnosis must be included in epidemiologic studies. PMID- 11055633 TI - Do parental factors affect male and female fertility? AB - There is little published evidence on parental characteristics and the fertility of their offspring of either sex. Maternal smoking has been reported to reduce fertility in both sexes and was also suggested to be relevant to the health of the male reproductive system on the basis of descriptive epidemiology. We undertook a cohort study based on a sample representative of the British population born in 1958 who have been followed up since birth. The outcome variable was time to pregnancy measured in months, up to age 33 years. Antecedent variables were the age of both parents; maternal smoking, height, prepregnancy body mass index, and parity; and paternal social class (manual/nonmanual labor). First births to cohort members were analyzed using a Cox logistic model for discrete "survival" times. A total of 1,714 and 2,587 values of time to pregnancy were available, respectively, for male and female cohort members. In the unadjusted analyses, all odds ratios were in the range 0.9-1.1, apart from the father's social class. In the adjusted analyses, this effect also disappeared. We conclude that the observed heterogeneity in biological fertility is unrelated to those characteristics of parents that we were able to analyze. PMID- 11055634 TI - Effect of home and hospital delivery on long-term cognitive function. AB - We examined the relation between place of birth and cognitive function in young adult life in a historical cohort study based upon birth data from the computerized Danish Medical Birth Registry and cognitive function as measured at time of drafting for military service in two Danish counties. The cohort included 4,296 Danish conscripts born between 1973 and 1976, 123 born at home and 4,173 born in hospital or at a birth clinic. Cognitive function was measured by the Boerge Prien test in men, 18 to 20 years of age. The highest possible score is 78. The mean Boerge Prien test score was 43.1 for conscripts born in specialized hospital departments, 2.4 higher for conscripts born in a birth clinic (95% confidence interval = 0.9-4.0), and 2.1 lower for conscripts born at home (95% confidence limits = -3.8 to -0.4) after adjusting for birth weight, length at birth, birth order, gestational age, maternal age, and marital and occupational status. Our findings raise the possibility that home birth can lead to lower cognitive function in adulthood; however, from our data we could not distinguish between planned and unplanned births at home. PMID- 11055635 TI - Community firearms, community fear. AB - To examine how perceptions of safety are influenced as more people in a community acquire firearms, we conducted a nationally representative random-digit-dial survey of 2,500 adults and asked whether respondents would feel more safe, less safe, or equally safe if more people in their community were to acquire guns. We used multivariable logistic regression to explore correlates of perceived safety while taking into account various confounders. Fifty percent of respondents reported that they would feel less safe if more people in their community were to own guns; 14% reported they would feel more safe. Women and minorities were more likely than were men and Whites to feel less safe as others acquire guns, with Odds ratios of 1.7 and 1.5, respectively. Our findings suggest that most Americans are not impervious to the psychological effects of guns in their community, and that, by a margin or more than 3 to 1, more guns make others in the community feel less safe rather than more safe. PMID- 11055636 TI - Perinatal medication as a potential risk factor for adult drug abuse in a North American cohort. AB - The aim of this study was to explore perinatal exposures to medications as risk factors for adult drug abuse. We compared 69 drug abusing subjects and their 33 non-abusing siblings with regard to history of labor pain analgesia during birth and other obstetric variables. Three or more doses of opiates or barbiturates at birth yielded an OR of 4.7 (95% CI = 1.0-44.1) for becoming a drug abuser after multiple perinatal drug exposure. PMID- 11055637 TI - Retrospectively sampled time-to-pregnancy data may make age-decreasing fecundity look increasing. European Infertility and Subfecundity Study Group. AB - The retrospective study based on pregnancy samples is the most commonly used method for studying time to pregnancy. Using a simple Monte Carlo simulation, this note focuses on a particular selection bias problem associated with this design: even if each woman's fecundity decreases with age, estimation of the effect of age may show the opposite trend. We recommend exercising increased caution in interpreting findings from pregnancy-based time-to-pregnancy studies. PMID- 11055638 TI - Sensitivity and predictive value positive measurements for public health surveillance systems. AB - Two important measurements for the evaluation of a public health surveillance system are sensitivity and predictive value positive (PVP). The computation of sensitivity and PVP for a public health surveillance system, however, can be complicated by the absence of an appropriate gold standard. In addition, there are few references for the computation of sensitivity and PVP for a surveillance system. To determine how these attributes of evaluation have been reported in epidemiologic literature, I review papers that report sensitivity and PVP for public health surveillance systems. Of the 31 papers that met selection criteria, 21 (68%) included either a reference for the computation or a definition of the attributes, whereas 18 (58%) reported both attributes. All 31 papers reported sensitivity, and among the 31 papers, 24 (77%) reported more than one sensitivity measurement. Among the 18 papers that reported at least PVP, 13 (72%) reported more than one PVP measurement. This review provides guidance in computing sensitivity and PVP for a public health surveillance system. PMID- 11055639 TI - The case for a comprehensive national campaign to prevent melanoma and associated mortality. PMID- 11055640 TI - Are pregnancies shorter in leap years? PMID- 11055641 TI - Epidemiology of suicide pacts in central Italy. PMID- 11055642 TI - Rise in mortality from coronary heart disease among post-World War II birth cohorts in Japan? PMID- 11055643 TI - Interval between pregnancies and risk of spontaneous abortions. PMID- 11055644 TI - Self-reported airway symptoms in a population exposed to heavy industrial pollution: what is the role of public awareness? PMID- 11055645 TI - Racism, poverty, abortion, and other reproductive outcomes. PMID- 11055646 TI - Significance of HIV-1 genetic subtypes. AB - HIV is the most significant new pathogen that emerged during the twentieth century. Since the recognition of AIDS in 1981, HIV has caused a worldwide epidemic. HIV-1 mutates extensively and shows high genetic diversity and thereby poses significant challenges for effective surveillance and disease control. At the beginning of the 1990s phylogenetic analyses of HIV-1 sequences from different sources of the world epidemic revealed that HIV-1 can be divided into different clades or subtypes. However, most of the knowledge from that time was based on information from western countries, where subtype B predominated. Important questions were raised about the possibility that genetic and phenotypic differences in HIV-1 may affect transmissibility, infectivity and pathogenicity, in addition to responses to therapy and vaccines. On this basis this study was initiated in 1994, and presented as a thesis in 1999. This paper gives an overview of the results from this thesis (based on 6 original papers) and the conclusions drawn. In summary, determination of the genetic subtype of HIV-1 probably has little value for routine clinical care of individual patients, but provides a powerful tool for monitoring changes in local and global transmission patterns. PMID- 11055647 TI - Chronic hepatitis C--Swedish experts' meeting recommends combination treatment. PMID- 11055648 TI - Mononuclear cells and cytokines in the cerebrospinal fluid of echovirus 30 meningitis patients. AB - The local immune response to echovirus 30 meningitis in children was investigated in 2 ways: using flow cytometry to examine the mononuclear cell and lymphocyte subsets and enzyme-linked immunosorbent assays to examine the cerebrospinal fluid (CSF) levels of cytokines, including monocyte chemoattractant protein-1 (MCP-1), interferon gamma (IFN-gamma) and interleukin-12 (IL-12). The numbers of macrophages in the CSF were increased, in particular during the early part of the acute stage. The levels of MCP-1, which is responsible for the accumulation of macrophages, as well as those of IFN-gamma and IL-12, which play important roles in the activation of macrophages and T helper (Th) 1 cells, respectively, were increased in the CSF of patients compared with the levels found in the controls. Likewise, numbers of activated CD4 + and CD8 + T lymphocytes were increased in the CSF. Since the ratio of CD4/CD8 correlated with the age of the patients, CD8 + T lymphocytes in the CSF might play a more important role in younger children. PMID- 11055649 TI - Analysis of HIV-1 genetic subtypes in Finland reveals good correlation between molecular and epidemiological data. AB - The molecular epidemiology of HIV-1 genetic subtypes was studied in a cross sectional sample collected from HIV-infected individuals living in Finland between 1988 and 1994 and compared with independently collected epidemiological data. Subtypes were determined by sequencing and phylogenetic analysis of the gag NCp7 and the env coding regions of PBMC provirus. Finnish viruses belonging to 7 subtypes were found. Two thirds (n = 70) of the sequences could be classified as subtype B, while others belonged to subtypes A, C, D, F and G and the circulating recombinant form AE(CM240) (n = 25). There were significant differences in gender distribution and mode-of-transmission between B-type infections and infections with the other subtypes. Most subtype B strains in Finland were associated with homosexual transmission and about half of these were acquired in Finland, while most individuals harbouring non-B infections indicated heterosexual transmission and direct or indirect contact with Africa or Southeast Asia. The heterogeneity of genetic subtypes in the country was in good agreement with the epidemiological data suggesting that a significant proportion of infections were imported. HIV-1 subtype determination may prove to be a valuable tool for providing objective epidemiological data. PMID- 11055650 TI - Rapid detection of dihydropteroate polymorphism in AIDS-related Pneumocystis carinii pneumonia by restriction fragment length polymorphism. AB - Sulpha agents, which act by inhibiting the enzyme dihydropteroate synthase (DHPS), are used widely for the treatment and prophylaxis of Pneumocystis carinii pneumonia (PCP). Recently, we have shown that mutations in the dihydropteroate synthase (DHPS) gene of Pneumocystis carinii f.sp hominis are associated with failure of sulpha prophylaxis and increased mortality in HIV-1 positive patients with PCP, suggesting that DHPS mutations may cause sulpha resistance. To facilitate detection of DHPS mutations we developed a restriction fragment length polymorphism (RFLP) assay, detecting mutations at codon 55 and 57 of the P. carinii DHPS gene. The RFLP-assay was compared with direct DNA sequencing on 27 PCP isolates from HIV-1 positive patients with a mixture of wildtype and mutant DHPS types. In all samples the RFLP-assay correctly identified wildtype or DHPS mutation at codon 55 or 57. Combined with DNA extraction by a Chelex-based method, this method can be performed within 1 d and allows a fast, cost-efficient and reliable method of detection of DHPS mutations in P. carinii. PMID- 11055651 TI - Aspergillus antigen in serum, urine and bronchoalveolar lavage specimens of neutropenic patients in relation to clinical outcome. AB - We have used a new, commercial enzyme-linked immunosorbent assay (ELISA, Platelia Aspergillus) to detect Aspergillus antigen in serum, urine and bronchoalveolar lavage (BAL) samples of 105 haematological patients who received empirical amphotericin B treatment for suspected fungal infection. 14% (60/419) of serum and 5% (18/373) of urine samples were positive. Ten-fold concentration of urine increased the number of positive samples to 31 (8%). The antigen was detected in 5 of 20 BAL samples, but fungal culture was negative in all of them. 22 patients had positive antigen test. Serum was positive in 17, urine in 7 and concentrated urine in 12 patients. Six patients had confirmed invasive aspergillosis. In all these patients, antigen was detected in serum, but urine was positive in only 2 patients. Patients whose antigen test turned negative during the amphotericin B treatment had significantly lower mortality than patients with persistently positive antigen test (2/10 vs. 8/8, p = 0.002). We conclude that Aspergillus galactomannan can be detected by ELISA in serum, urine and BAL samples of haematological patients, but the higher sensitivity of serum testing makes it preferable for screening. Disappearance of the antigen during antifungal therapy seems to correlate with good, and persistence with poor, clinical outcome. PMID- 11055652 TI - Streptococcus pneumoniae septic arthritis in adults. AB - Septic arthritis is a rarely reported manifestation of disease due to Streptococcus pneumoniae. We have reviewed our recent experience of this disease in 14 adult patients. Common features in patients with S. pneumoniae septic arthritis included advanced age (median=63 y), pre-existing joint disease (6/14), large joint disease (14/14), polyarthritis (6/14), and associated meningitis, pneumonia or both (6/14). Two patients with septic arthritis and meningitis, and another with Down's syndrome and sleep apnoea, died during treatment. In the remaining 11 patients, treatment for at least 19 d, predominantly with intravenous benzyl penicillin, plus joint lavage, resulted in cure. PMID- 11055653 TI - Recurrence of urinary tract infections in adult patients with community-acquired pyelonephritis caused by E. coli: a 1-year follow-up. AB - In a prospective study, 42 women were followed for recurrence of urinary tract infections (UTIs) for 1 y after an index episode of community-acquired pyelonephritis caused by Escherichia coli. Altogether, 26 repeat episodes were detected. Of these, 20 occurred at least 1 month after the index episode and were regarded as recurrences. In all, 40%, (17 of 42) of the women had recurrences. An earlier history of UTI increased the risk of recurrence: 52%, of the 29 women with previous UTI had at least 1 recurrence, compared with 15%, of the 13 patients without previous UTI. E. coli caused the majority (73%) of the recurrences. Genotype comparisons by RAPD-PCR analysis between E. coli isolates from a patient showed that 75%. of the original and recurrent strains were genetically non-identical. Of the 54 E. coli strains, 42 were carrying genes coding for G adhesins of P fimbriae: 40 isolates carried class II, I class III and 1 carried both class II and III G adhesin genes. Each of the virulence associated factors (genes for G adhesins, MRHA, haemolysin, type 1C fimbriae, and O and K antigens) was evenly distributed among E. coli isolates of index episodes, independent of the recurrences. The index isolates, however, had more virulence-associated factors than did the isolates from the recurrences which were mainly due to lower UTIs. PMID- 11055654 TI - Mycobacterium tuberculosis infections after renal transplantation. AB - The incidence of tuberculosis was found to be 5.8% (16/274) in 274 kidney graft recipients in our centre between 1986 and 1998. The kidney recipients were evaluated retrospectively. A total of 51 recipients received isoniazid prophylaxis for 6 months. The prevalence of tuberculosis was found similar (6% vs. 8.8%, p = 0.15) between recipients with prophylaxis and no prophylaxis. Eight patients were recipients of cadaveric donor kidneys and 8 were recipients of living donor kidneys. Lungs were the most frequently affected site, as in the normal population. M. tuberculosis grew in 7 patients. In 5 patients, M. tuberculosis was also detected on direct microscopy and polymerase chain reaction. In 4 patients, diagnosis was made on clinical grounds and later confirmed by positive response to therapy. In 8 patients, invasive procedures were performed for diagnosis. Five patients had miliary tuberculosis at the time of diagnosis. In 3 patients dissemination occurred during follow-up. Nine patients responded to anti-tuberculous therapy while still preserving their graft function, 1 patient rejected the graft while under treatment and returned to haemodialysis. Five patients (31%) died. Since the risk of dissemination of tuberculosis is high in these patients, anti-tuberculous therapy should be started whenever clinical findings suggestive of tuberculosis are present, even in the absence of any microbiological and/or histological evidence. PMID- 11055655 TI - Mycobacterium tuberculosis drug resistance in Turkey, 1976-97. AB - Drug-resistant tuberculosis is increasing day by day and is a significant threat to tuberculosis control because there are few drugs effective against Mycobacterium tuberculosis. This study evaluates the resistance of the microorganism to primary anti-tuberculosis drugs over the 21-y period 1976-97. Records from the bacteriology laboratory of the Department of Chest Diseases and Tuberculosis, Ankara University Medical Faculty were evaluated retrospectively. Among 3,418 mycobacteria strains, 3,319 (97.1%,) M. tuberculosis were isolated and their susceptibility was examined by the proportion method in Loewenstein Jensen medium. It was found that 60.8% of isolated strains were susceptible, whereas 39.2%, were resistant to at least one drug. Multi-drug resistant tuberculosis (MDR-TB) was found in 194 (5.8%) materials. Over the 21-y period studied, total resistance to isoniazid (INH), rifampicin (RF) and streptomycin (SM) were determined as 10.5, 6.9 and 7.0%, respectively. It was also observed that the resistance rates to INH or SM increased, whereas resistance to RF was not changed within this period. While resistance to the 2-drug combination RF+SM increased, resistance to INH+SM decreased significantly. There was no change in resistance to the 2-drug INH + RF or 3-drug INH + RF + SM combinations in the same period. In conclusion, combined therapy is still useful and available for the treatment of resistant tuberculosis, and INH should be included in the chemotherapeutic regimen even if high resistance rates are shown to exist. PMID- 11055656 TI - Plasmapheresis as part of the treatment for septic shock. AB - Plasmapheresis is one of the methods which has been tried in the effort to influence the course of severe sepsis with septic shock and to improve survival rates. This is a retrospective study of 17 consecutive patients with septic shock who were treated with acute plasmapheresis. Nine out of 16 patients with verified etiology suffered from infections with Gram-positive bacteria. Three (18%) of the treated patients died. The estimated mortality rate based on the patients' initial APACHE II scores was 62%. No serious side-effects of treatment were observed. This study indicates that plasmapheresis is a safe treatment for patients with septic shock and has a positive effect on survival. Prospective, randomized studies would be valuable to determine the role of plasmapheresis in the management of patients with septic shock. PMID- 11055657 TI - C-reactive protein and fever in neutropenic patients. AB - The predictive value of daily C-reactive protein (CRP) monitoring to distinguish causes of fever in neutropenic patients was studied retrospectively. A total of 143 fever episodes during 113 consecutive hospitalizations were studied in 71 patients who had been referred for chemotherapy or haemopoietic stem cell transplantation (HSCT). There were, on average, 1.3 fever episodes per hospital stay, attributed to: infection (55, 27 invasive bacterial, 5 fungal, 3 viral and 20 probable infections); acute graft vs host disease (GvHD) (20); drugs (22); transfusions (7); and not attributable (39). 130 (91%) fever episodes were accompanied by a rise in CRP, 6 (4%) episodes were fatal. Maximal CRP levels (CRPmax) and maximal temperature (Tmax) were higher in invasive bacterial infections than in aGvHD and higher in aGvHD than in drug- or transfusion-related fever (p < 0.0001). Temperature and CRP rose in parallel. A total of 16 patients developed grade II-IV aGvHD by day 11 (9-21) (median, range) after allogeneic HSCT. Acute GvHD was preceded by fever for 3 d (1-7), and by CRP increase for 5 d (0-15) (p < 0.0001). CRP monitoring may be useful to distinguish between causes of fever. Very high CRP levels tend to be associated with invasive bacterial infections. CRP is not an early warning sign. An increase in CRP and fever may precede other clinical manifestations of aGvHD. PMID- 11055658 TI - Risk and patterns of bacteraemia after splenectomy: a population-based study. AB - During a period in which vaccination of splenectomized patients has been recommended, we analysed the patterns of severe post-splenectomy infections (i.e. bacteraemia or meningitis) in a defined population-based cohort. A total of 561 patients undergoing splenectomy were identified during 1984-93 in a Danish county, and the 538 eligible patients were followed for 1731 person-years. After splenectomy, 38 patients contracted a bacteraemia, of which 45% occurred within 30 d (i.e. during the postoperative period). No cases of meningitis were found. Among splenectomized patients the incidence rate of bacteraemia was 2.3 per 100 person-years at risk. Beyond the postoperative period we found an 8-fold increased risk of bacteraemia. Enterobacteria were the predominant cause (45%), and only 1 case due to Streptococcus pneumoniae was recorded. 89 (17%) died during the postoperative period, and the overall mortality rate was 18.4 per 100 person-years at risk. In all, 60% of the patients had been given a pneumococcal vaccination, and a Cox proportional hazard regression model showed that vaccination significantly reduced the risk of bacteraemia of any cause beyond the postoperative period. We conclude that splenectomy increases the risk of severe infections, and that vaccinated patients carry a lower risk of infection than non vaccinated ones. PMID- 11055659 TI - Surveillance of cases of meningococcal disease associated with military recruits studied for meningococcal carriage. AB - Through a 14-months extended surveillance of meningococcal disease in Denmark, all 322 notified cases were investigated for possible connection with a military camp where 3 cohorts of recruits (n = 1069) were studied prospectively for meningococcal carriage. One case occurred in a recruit who was a constant non carrier during the preceding 3 months. The invasive Neisseria meningitidis B:1:P1.1,7 strain was isolated from the pharynx only in 3 out of 17 room-mates (18%); the strains were identical as assessed by genotyping (PFGE and ribotyping). Two civilian cases outside the camp had direct contact with 2 recruits, but neither these 2 nor other recruits in the relevant divisions carried the invasive strains on any occasion. Six civilian cases had marginal relationship with the camp, but no contact with the recruits. In conclusion, pheno- and genotyping concordantly demonstrated a high carriage rate of the invasive strain among the room-mates to a recruit with meningococcal disease. Transmission to the patient most likely occurred shortly before onset of illness. The extended surveillance did, however, not identify any unexpected epidemiological links and restriction of antibiotic chemoprophylaxis to household/sleeping/kissing contacts in sporadic cases of meningococcal disease seems appropriate and relevant. PMID- 11055660 TI - Human leptospirosis in Denmark 1970-1996: an epidemiological and clinical study. AB - Epidemiological and clinical aspects of 118 laboratory confirmed cases of human leptospirosis in Denmark from 1970 to 1996 were reviewed. Icterohaemorrhagiae (72%) and sejroe (20%) were the most frequent serovars. The incidence of leptospirosis was 0.09/100,000 inhabitants/y. 93% of the patients were 18-64 y old, 90% were men and 72% of the cases occurred from July to November. Occupational exposure was present in 63% (74/118) of the cases (41% fish farmers, 28%, farmers). Eight percent of the patients had travelled abroad, 7% had been exposed to sewers and 4%, had been exposed through recreational activities (fishing). Initial symptoms were generally non-specific. Weil's disease developed in 63%, of the patients, more often in patients infected with the serovar icterohaemorrhagiae (73%) compared to patients infected with serovars sejroe or saxkoebing (25%). The fatality rate was 7%, all due to icterohaemorrhagiae. Though a rare disease in Denmark, leptospirosis should be considered in certain risk groups as a possible diagnosis in patients with acute febrile illness. PMID- 11055661 TI - Temporal risk assessment for Lyme borreliosis in Denmark. AB - The prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus nymphs in Denmark was found to be approximately 5%. The mean abundance of infected nymphs varied from 0.3 to 4.4 per 100 m2 according to site. The seasonal occurrence of infected nymphs in a beech forest coincided with seasonal distribution of neuroborreliosis cases. In order to establish a working hypothesis, it was assumed that the availability of habitats and human habitat preferences is one of the factors leading to low number of neuroborreliosis cases in the spring. In addition, this paper gives a description of the neuroborreliosis cases in Denmark in the period 1985-97 and offers a possible explanation for the variation in cases. The explanation is based on an assessment of tick density, which by comparison with the number of neuroborreliosis cases provides information on the infectivity of ticks. The model suggests that high temperatures and low precipitation in the autumn is essential for the transmission of B. burgdorferi sensu lato to reservoir hosts or development of B. burgdorferi sensu lato within ticks, which secures high tick infectivity in the following season. PMID- 11055662 TI - Spatial risk assessment for Lyme borreliosis in Denmark. AB - A study of nymphal and adult Ixodes ricinus density was performed in well-defined spruce and beech forest habitats with different levels of roe deer abundance and soil water capacity. In 35 habitats, a total of 489 larvae, 1,611 nymphs and 193 adult I. ricinus ticks were collected. Tick density was found to be influenced by roe deer abundance and soil water capacity. Based on this evaluation, a model predicting increasing number of ticks with increasing roe deer abundance and soil water capacity was suggested. A total of 1,045 nymphs and 106 adult ticks were tested for infection with Borrelia burgdorferi sensu lato Of these, 53 nymphs and 6 adults were found to be infected, leading to an general infection rate of 5% and 6% for nymphs and adults, respectively. Prevalences of Borrelia burgdorferi sensu lato in nymphal I. ricinus were found to be independent of roe deer abundance and soil water capacity. The correlation between human neuroborreliosis incidence and the estimated number of I. ricinus based on roe deer abundance and soil water capacity was examined. Differences in human neuroborreliosis incidence were found to correspond with the expected spatial differences in tick density in 12 counties in Denmark. PMID- 11055663 TI - Isolation and characterization of a 31 kDa mycobacterial antigen from tuberculous sera and its identification with in vitro released culture filtrate antigen of mtb H37Ra bacilli. AB - Antigens released in vivo are of considerable interest in the immunodiagnosis of infectious diseases. Circulating antigen was isolated from bacteriologically confirmed tuberculous sera by ammonium sulphate precipitation. The protein fraction between 36%, and 75%, ammonium sulphate was reactive with tuberculosis (TB) sera showing the presence of circulating tubercular antigen (CTA). Fractionation of CTA on ultrogel AcA 34 gel filtration column gave 3 protein fractions CTA1, CTA2 and CTA3. CTA2 showed maximum antigenic activity by sandwich enzyme-linked immunosorbent assay (ELISA). SDS-PAGE fractionation and seroreactivity studies showed the presence of highly reactive tubercular antigen in CTA2-7 protein fraction by sandwich ELISA. Further fractionation of CTA2-7 on cation exchange fast-protein liquid chromatography (FPLC) gave 4 antigenic fractions, of which CTA2-7D was seroreactive similar to 31 kDa antigen (ESAS-7F) isolated from in vitro culture medium. Furthermore, CTA2-7D could inhibit binding of in vitro released ESAS-7F to affinity purified antibodies in inhibition ELISA. CTA2-7D antigen may be used as a target antigen in confirming active tubercular infection. Biochemical characterization showed circulating antigen CTA2-7D to be a lipoglycoprotein is released in vivo. ESAS-7F as a glycoprotein is released in vitro culture. PMID- 11055664 TI - Thrombotic microangiopathy as a cause of dilated cardiomyopathy in HIV-infected patients. AB - Thrombotic microangiopathy is a rare cause of dilated cardiomyopathy. Both entities have been described separately in the setting of HIV infection. However, no patient with these 2 conditions has been reported to date. We report here 2 HIV-infected patients with dilated cardiomyopathy seemingly caused by thrombotic microangiopathy. PMID- 11055665 TI - Acute fascioliasis with multiple liver abscesses. AB - Human fascioliasis is distributed worldwide with several foci of high endemicity. Being a rare disease in Europe, we describe here a case in the initial hepatic phase of the disease. Therapeutic and, with reference to the 2 distinct stages of disease, diagnostic standards are discussed. PMID- 11055666 TI - Successful treatment with liposomal amphotericin B of an intraabdomianl abscess due to Candida norvegensis associated with a Gore-Tex mesh infection. AB - There are few reports of severe infections caused by Candida norvegensis. We here describe a case of C. norvegensis-associated intraabdominal abscess and Gore-Tex mesh-associated infection, successfully treated with liposomal amphotericin B and removal of the mesh. This is, to our knowledge, the first report of C. norvegensis causing this type of infection. PMID- 11055667 TI - Brain abscess caused by methicillin-resistant Staphylococcus aureus. AB - A Swedish tourist was admitted to a Cuban hospital due to epileptic seizures caused by brain tumors. Upon return to Sweden and admission to our hospital, methicillin-resistant Staphylococcus aureus (MRSA) was isolated. He was later considered to be free of MRSA but then developed a brain abscess from which MRSA was isolated. PMID- 11055668 TI - Pyopneumothorax: a complication of Streptococcus pyogenes pharyngitis. AB - A 20-y-old African-American female with Streptococcus pyogenes pharyngitis presented with tension pyopneumothorax. Her illness began with fever and sore throat that persisted for several days. She then developed a left neck swelling, followed by difficult swallowing and cough. Subsequently, she developed shortness of breath that became severe. On physical examination fever (39.2 degrees C), exudative pharyngitis, tenderness and swelling in the left anterior cervical area were noted. Chest X-ray revealed left side pneumothorax, air-fluid level and near complete collapse of the left lung with displacement of the heart and trachea to the right. Computed tomography scan of the neck revealed swelling and enhancement of the sternocleidomastoid muscle with loculated fluid collection, inflammation in the left anterior medial neck displacing the trachea extending into the mediastinum and the left apex. Thoracentesis revealed purulent fluid; Gram stain showed Gram-positive cocci in chains; culture yielded pure growth of Streptococcus pyogenes. She was treated with high dose penicillin, several chest tubes and intra-pleural injections of streptokinase with gradual resolution. This complication has not been described previously in Streptococcus pyogenes pharyngitis. PMID- 11055669 TI - Vertebral osteomyelitis due to Pasteurella aerogenes. AB - A case of C6-C7 vertebral osteomyelitis due to Pasteurella aerogenes in a previously healthy 62-y-old man in the absence of any history of animal exposure, debilitating disease or immunosuppression is reported. Culture testing of biopsy samples of the vertebral body using the panels and database of the BBL Crystal enteric/non-fermenter system revealed that the infecting bacterium was P. aerogenes. Treatment with cloxacillin and gentamicin was followed by resolution of bone infection on serial follow-up magnetic resonance imaging scans. Pasteurellae are primarily animal pathogens but are capable of producing a variety of local and systemic diseases in humans. PMID- 11055670 TI - Gonococcal meningitis and intra-abdominal abscess in the presence of a ventriculoperitoneal shunt. AB - We report a case of gonococcal meningitis with an intraabdominal abscess in a 19 y-old woman, who had a ventriculoperitoneal shunt after craniotomy for astrocytoma. Percutaneous drainage was performed for the intraabdominal abscess. Ceftriaxone was administered with the ventriculoperitoneal shunt removal. The clinical course improved, but visual impairment last for 3 months following treatment. PMID- 11055671 TI - Mycobacterium interjectum: a new pathogen in humans? AB - Molecular genetic techniques have increased the number of species recognized within the genus Mycobacterium. The clinical significance of these is uncertain and their pathogenic potential has still to be proven. We describe here a case of mycobacterial lymphadenitis in a Swedish boy, which supports the role of M. interjectum as a human pathogen. PMID- 11055672 TI - Regression of HIV-associated progressive encephalopathy of childhood during HAART. AB - HIV-associated progressive encephalopathy of childhood is characterized by impaired brain growth, decline in cognitive and neurobehavioral performances, and progressive motoric dysfunction The diagnosis is based on neurological examination, neuropsychological assessment and cerebral CT or MR imaging. While the importance of early use of antiretroviral combination therapy has been emphasized, limited data exist as to the effect of protease inhibitors in children with HIV-associated encephalopathy. We describe the effect of 3-drug antiretroviral combination therapy, including the protease inhibitor nelfinavir, in a 7-y-old girl with vertically acquired HIV infection and late onset progressive encephalopathy. PMID- 11055673 TI - Amantadine monotherapy of chronic hepatitis C patients infected with genotype lb. AB - We aimed to test the efficacy of amantadine in chronic hepatitis C (CHC) patients infected with genotype b. Twenty patients completed treatment with amantadine HCl, 100 mg b.i.d., for 6 months. Non-sustained biochemical improvement was observed without loss of HCV-RNA. We conclude that amantadine monotherapy is not effective in CHC. PMID- 11055674 TI - Cervical spinal epidural abscess due to group B streptococcus in a previously healthy elderly male. PMID- 11055675 TI - Shigella flexneri bacteremia in a middle-aged immunocompetent woman. PMID- 11055676 TI - A teratological study of lincosamides. PMID- 11055677 TI - Biliary excretion of amphotericin B deoxycholate and amphotericin B lipid complex. PMID- 11055678 TI - Klebsiella ozaniae empyema complicating longstanding oleothorax. PMID- 11055679 TI - Syncope. AB - Syncope is a common clinical presentation. Although most commonly benign, it may herald a pathology with a poor prognosis. The work-up of syncope includes a careful history, physical examination, electrocardiogram, risk stratification, and appropriately directed testing. The key factor in the investigation of syncope is the presence (or absence) of structural heart disease or an abnormal electrocardiogram. The most useful investigation in unexplained syncope with a normal heart is the tilt table test for evaluating predisposition to neurocardiogenic (vasovagal) syncope. In the setting of structural heart disease or an abnormal electrocardiogram, electrophysiologic studies play a more important role. The utility of noninvasive cardiac monitoring for symptom-rhythm correlation may be limited by infrequent symptoms. The availability of external and implantable loop recorders allows prolonged periods of monitoring to increase diagnostic yield. The management of patients with syncope may be complex. Early referral to a cardiac electrophysiologist is warranted in patients who are at high risk. PMID- 11055680 TI - Bradyarrhythmias, temporary and permanent pacing. AB - Bradycardia is common in critical care units. It may be transient, asymptomatic and of little consequence, or life-threatening. Bradycardia may result from abnormalities of the sinus node, atrioventricular node, or the His-Purkinje system. It may also be precipitated by drug effects or enhanced vagal tone. Proper diagnosis is pivotal to determining prognosis and management. Temporary and permanent pacing is now readily available, markedly improving the morbidity and mortality associated with bradyarrhythmias. PMID- 11055681 TI - Supraventricular tachycardia: implications for the intensivist. AB - The critical care physician must have a keen awareness of supraventricular tachycardia patterns, mechanisms, precipitants, and treatment. Although long-term management of most forms of supraventricular tachycardia lies primarily in the realm of the cardiac electrophysiologist, the intensivist must be proficient at acute arrhythmia therapy. Expertise in electrocardiography, pharmacokinetics, and pharmacodynamics is essential. Careful assessment of hemodynamics and prudent bedside clinical acumen help assure optimal patient outcomes. PMID- 11055682 TI - Cardiac surgery: postoperative arrhythmias. AB - Arrhythmias are common after surgery, particularly after cardiac surgery. Atrial fibrillation is the most common arrhythmia encountered postoperatively, although ventricular arrhythmias and conduction disturbances can also occur. Older age is the most consistent predictor of postoperative atrial arrhythmias. beta adrenergic blockers, amiodarone, and sotalol are the most effective at preventing postoperative atrial fibrillation. Sustained ventricular arrhythmias in the recovery period after cardiac surgery warrant aggressive therapy, usually with an implantable cardioverter-defibrillator in the absence of reversible causes. Postoperative, nonsustained ventricular tachycardia in the setting of left ventricular dysfunction and ischemic coronary disease also usually warrants risk stratification and possible treatment, often with electrophysiologic testing and implantation of an implantable cardioverter-defibrillator, if sustained ventricular arrhythmias are induced. Transient bradyarrhythmias may be managed with temporary pacing wires placed at surgery, but significant and persistent atrioventricular block or sinus node dysfunction can occur and indicate a need for permanent pacing. PMID- 11055683 TI - Noncardiac surgery: postoperative arrhythmias. AB - Postoperative arrhythmias are common and represent a major source of morbidity after both cardiac and noncardiac surgical procedures. Postoperative dysrhythmias are most likely to occur in patients with structural heart disease. The initiating factor for an arrhythmia in a given patient after surgery is usually a transient insult, such as hypoxemia, cardiac ischemia, catecholamine excess, or electrolyte abnormality. Management includes correction of these imbalances and medical therapy directed at the arrhythmia itself. The physiologic impact of arrhythmias depends on arrhythmia duration, ventricular response rate, and underlying cardiac function. Similarly, urgency and type of treatment is determined by the physiologic impact of the arrhythmia, as well as by underlying clinical status. The purpose of this review is to provide current concepts of diagnosis and acute management of arrhythmias after noncardiac surgery. A systematic approach to arrhythmia diagnosis and evaluation of predisposing factors is presented, followed by consideration of specific bradyarrhythmias and tachyarrhythmias in the postoperative setting. PMID- 11055684 TI - Ischemia, metabolic disturbances, and arrhythmogenesis: mechanisms and management. AB - The development of ventricular arrhythmias is often a consequence of the interaction between structural abnormalities of the heart and transient disturbances in the electrophysiologic milieu. The critically ill patient is particularly susceptible to arrhythmias given the metabolic, ischemic, and neurohormonal stressors present in the intensive care unit. The significance of ventricular arrhythmias in the acute care setting is related to the presence of reversible causes and the extent of underlying heart disease. Long-term management of these patients is guided by an assessment of the risk for recurrent arrhythmias and the degree of left ventricular systolic dysfunction. In the absence of a reversible cause, symptomatic sustained arrhythmias are usually treated with an implantable cardioverter-defibrillator, a therapy that improves survival in this patient population. In many cases, however, proper long-term management of patients with ventricular arrhythmias is less clear, and the approach must be guided by a thorough understanding of the pathophysiology and the fundamental mechanisms of arrhythmogenesis. PMID- 11055685 TI - Antiarrhythmic agents and proarrhythmia. AB - The Vaughn Williams classification divides antiarrhythmic agents into four groups according to their effects on various ion channels. Class I agents block sodium channels and are subdivided into three groups. The use of class Ia agents is gradually on the decline, secondary to lack of a favorable risk/benefit ratio. Class Ib agents include lidocaine, which is extensively used for the acute treatment of ventricular tachyarrhythmias. Class Ic drugs are not advisable for patients with structural cardiac abnormalities secondary to a high risk of proarrhythmia. They are mainly used for supraventricular tachyarrhythmias. beta blockers form class II. Class III agents, such as amiodarone and sotalol, prolong action potential duration and repolarization and are among the most widely used antiarrhythmics. They are the subject of active research, and newer agents are being developed. Calcium-channel blockers are grouped under class IV. Digoxin and adenosine have unique antiarrhythmic properties, which can be useful in the management of selected patients. All antiarrhythmic drugs have the potential to provoke arrhythmias and, therefore, should be used with caution. The risk of proarrhythmia is increased in patients with abnormal cardiac substrate, with electrolyte abnormalities, and during drug initiation. Correction of electrolyte imbalance and prevention of bradycardia while the drug is metabolized and/or excreted are the cornerstones of proarrhythmia management. PMID- 11055686 TI - Management of malignant ventricular arrhythmias and cardiac arrest. AB - Sudden cardiac death continues to be a major health problem in the United States, accounting for approximately 400,000 deaths per year. The last 10 yrs have seen major advances in the primary and secondary prevention of this problem. In patients who have survived an episode of cardiac arrest, the AVID study conclusively established the superiority of the implantable cardioverter defibrillator over empiric amiodarone. For patients with recurrent hemodynamically destabilizing ventricular tachycardia and ventricular fibrillation, intravenous amiodarone has emerged as a potent therapeutic agent, especially when other agents such as lidocaine and procainamide have not been effective. Finally, recent work has focused on the risk stratification of patients for sudden cardiac death. Both the MADIT and MUSTT studies suggest that patients with coronary artery disease, reduced ejection fraction, and nonsustained ventricular tachycardia who are inducible to a sustained ventricular arrhythmia at electrophysiology testing have improved survival with an implantable cardioverter defibrillator. PMID- 11055687 TI - Direct current cardioversion: indications, techniques, and recent advances. AB - Direct current cardioversion/defibrillation is an important part of the intensivist's armamentarium. Emergent application may be lifesaving. Elective cardioversion should be used cautiously, with attention to patient selection and proper techniques. Repetitive, futile attempts at direct current cardioversion should be avoided. Reducing or eliminating arrhythmia precipitants may be safer and more effective than this more dramatic intervention. PMID- 11055688 TI - Emergencies related to implantable cardioverter-defibrillators. AB - Implantable cardioverter-defibrillators (ICDs) have become the dominant therapeutic modality for patients with life-threatening ventricular arrhythmias. ICDs are implanted using techniques similar to standard pacemaker implantation. They not only provide high-energy shocks for ventricular fibrillation and rapid ventricular tachycardia, but also provide antitachycardia pacing for monomorphic ventricular tachycardia and antibradycardia pacing. Devices incorporating an atrial lead allow dual-chamber pacing and better discrimination between ventricular and supraventricular tachyarrhythmias. Intensivists are increasingly likely to encounter patients with ICDs. Electrosurgery can be safely performed in ICD patients as long as the device is deactivated before the procedure and reactivated and reassessed immediately afterward. Prompt and skilled intervention can prove to be life-saving in patients presenting with ICD-related emergencies, including lack of response to ventricular tachyarrhythmias, pacing failure, and multiple shocks. Recognition and treatment of tachyarrhythmia can be temporarily disabled by placing a magnet on top of an ICD. The presence of an ICD should not deter standard resuscitation techniques. Multiple ICD discharges in a short period of time constitute a serious situation. Causes include ventricular electrical storm, inefficient defibrillation, nonsustained ventricular tachycardia, and inappropriate shocks caused by supraventricular tachyarrhythmias or oversensing of signals. ICD system infection requires hardware removal and intravenous antibiotic therapy. Deactivation of an ICD with the consent of the patient or relatives is reasonable and ethical in terminally ill patients. PMID- 11055689 TI - Electroosmotic fluid motion and late-time solute transport for large zeta potentials AB - Analytical and numerical methods are employed to determine the electric potential, fluid velocity, and late-time solute distribution for electroosmotic flow in a tube and channel at zeta potentials that are not necessarily small. The electric potential and fluid velocity are in general obtained by numerical means. In addition, new analytical solutions are presented for the velocity in a tube and channel in the extremes of large and small Debye layer thickness. The electroosmotic fluid velocity is used to analyze late-time transport of a neutral nonreacting solute. Zero- and first-order solutions describing axial variation of the solute concentration are determined analytically. The resulting expressions contain eigenvalues representing the dispersion and skewness of the axial concentration profiles. These eigenvalues and the functions describing transverse variation of the concentration field are determined numerically using a shooting technique. Results are presented for both tube and channel geometries over a wide range of the normalized Debye layer thickness and zeta potential. Simple analytical approximations to the eigenvalues are also provided for the limiting cases of large and small values of the Debye layer thickness. PMID- 11055690 TI - Activated ion electron capture dissociation for mass spectral sequencing of larger (42 kDa) proteins. AB - In previous studies, electron capture dissociation (ECD) has been successful only with ionized smaller proteins, cleaving between 33 of the 153 amino acid pairs of a 17 kDa protein. This has been increased to 99 cleavages by colliding the ions with a background gas while subjecting them to electron capture. Presumably this ion activation breaks intramolecular noncovalent bonds of the ion's secondary and tertiary structure that otherwise prevent separation of the products from the nonergodic ECD cleavage of a backbone covalent bond. In comparison to collisionally activated dissociation, this "activated ion" (AI) ECD provides more extensive, and complementary, sequence information. AI ECD effected cleavage of 116, 60, and 47, respectively, backbone bonds in 29, 30, and 42 kDa proteins to provide extensive contiguous sequence information on both termini; AI conditions are being sought to denature the center portion of these large ions. This accurate "sequence tag" information could potentially identify individual proteins in mixtures at far lower sample levels than methods requiring prior proteolysis. PMID- 11055691 TI - Capillary electrophoresis--matrix-assisted laser desorption/ionization time-of flight mass spectrometry using a vacuum deposition interface. AB - An improved vacuum deposition interface for coupling capillary electrophoresis with MALDI-TOF MS has been developed. Liquid samples consisting of analyte and matrix were deposited on a moving tape in the evacuated source chamber of a TOF mass spectrometer, enabling 24 h of uninterrupted analysis. The vacuum deposition procedure was compared with the dried-droplet method, and it was found that vacuum deposition generated significantly more reproducible signal intensity, eliminating the need for "sweet spot" searching. A concentration detection limit in the low-nanomolar range has been achieved with a low-attomole amount of sample consumed per spectrum. In addition, ion suppression caused by hydrophobicity differences in the analytes was reduced. To minimize ion suppression further, separation prior to MALDI MS analysis was employed. The performance of capillary electrophoresis (CE)-MALDI-TOF MS using the vacuum deposition interface was evaluated with a peptide mixture injected at low-femtomole levels. All peptides were baseline resolved with separation efficiencies in the range of 250000-400000 plates/m (2-3-s band half-width), demonstrating the high separation efficiency of the CE-MALDI MS coupling. A fast (approximately 40 s) CE separation of a mixture of angiotensins was found to reduce significantly ion suppression and enable trace level detection. It was also shown, for the analysis of an enolase digest, that sequence coverage of 65% was obtained using CE separation compared to 52% using step-elution solid-phase extraction and 44% in the control experiment using an unseparated mixture. PMID- 11055692 TI - Optical patternation: a technique for three-dimensional aerosol diagnostics AB - A novel technique based on optical patternation is described for three dimensional diagnostic studies of aerosols used in analytical spectroscopies. The aerosol is illuminated with a thin laser light sheet to capture images of the fluorescence and Lorenz-Mie light-scattering signals from the aerosol field with a charge-coupled detector. These measurements allow for the rapid and nonintrusive elucidation of two-dimensional spray structures, planar mass distributions, and spatial droplet size distributions. The ratio of the fluorescence image to the Lorenz-Mie image is then utilized to construct a spatially resolved map of the volume-to-surface area mean of the aerosol (Sauter mean diameter). Three-dimensional maps of spray structure, mass distribution, and droplet size distribution are obtained for the entire aerosol field by image stacking. The technique is applied to the measurement of the droplet size over the aerosol field at distances of 5-30 mm from the nebulizer tip where droplet sizes ranged from 6 to 12 microm for a direct injection high efficiency nebulizer used in inductively coupled plasma spectrometries. PMID- 11055693 TI - On-line concentration and separation of proteins by capillary electrophoresis using polymer solutions. AB - Proteins were separated in 0.6% poly(ethylene oxide) (PEO) solutions using a capillary filled with buffers prior to analysis and were detected by laser induced native fluorescence using a pulsed Nd:YAG laser. PEO solutions entered the capillary by electroosmotic flow (EOF) during the separation. The composition and concentration of the buffer affected the adsorption of PEO molecules on the capillary surface and, consequently caused changes in the EOF. Short separation times (< 7 min) were achieved on a sample solution of five proteins in a 0.6% PEO solution containing 5 microg/mL ethidium bromide using a capillary pre-filled with 100 mM TRIS-borate (TB) buffers (pH 10,0). We also extended this method for on-line concentration and separation of proteins. Proteins dissolved in low conductivity media stacked in both TB buffers and in PEO solutions. The peak height was proportional to the injection volume up to 2.1 microL using an 80-cm capillary filled with 400 mM TB buffers. Using large injection volumes (2.1 microL), we achieved a limit of detection (S/N = 3) of 31 pM for carbonic anhydrase, which was a 1696-fold sensitivity enhancement compared to a conventional injection method (1 kV for 10 s). In high-conductivity media (urine matrix), stacking occurred at the boundary between the sample zone and PEO solutions. A urine sample without any pretreatment was analyzed, and after stacking, several peaks were detected. Spiking the urine sample with human serum albumin (HSA) affected the fluorescent intensity of some analytes as a result of interaction with HSA. PMID- 11055694 TI - A capillary electrophoretic reactor with an electroosmosis control method for measurement of dissociation kinetics of metal complexes AB - The solvolytic dissociation rate constants of 1:2 complexes of Al3+ and Ga3+ with an azo dye ligand, 2,2'-dihydroxyazobenzene-5,5'-disulfonate (DHABS, H2L2-), have been evaluated with a capillary electrophoretic reactor (CER) system. This CER system is based on the fact that metal complexes encounter an overwhelming force to dissociate when apart from the ligand by CE resolution. Treatment of a capillary with a slightly acidic buffer solution, e.g., pH 5, reduces the double layer potential (zeta) of the inner silica wall. Owing to slow relaxation of the deprotonation equilibria of superficial silanol groups known as the pH hysteresis, this zeta potential can be actually retained during the electrophoresis of the metal complexes in question with a neutral buffer at pH 7.0. This method enables one to manipulate migration times, namely, residence times in a capillary tube, from 5 to 90 min, depending on the prescribed conditioning pH, without changing any other operation conditions such as buffer composition and electric field strength. The excellent performance of the CER is exemplified by the accurate estimation of the dissociation degree of the complexes. The dissociation degree-time profiles for the complexes are quantitatively described using both internal and external standards; the very inert complex of [Co(III)L2]5- for the peak signal standardization and methyl orange for the injection volume correction. The solvolytic dissociation rate constants of the 1:2 complexes of Al3+ and Ga3+ ions with DHABS [AlL2]5- and [GaL2]5- into the 1:1 ones have been determined as (4.9+/-1.0) x 10(-4) and (3.7+/-0.3) x 10(-3) s(-1) at 303 K, respectively. PMID- 11055695 TI - Capillary electrophoresis with an integrated on-capillary tubular detector based on a carbon sol-gel-derived platform AB - An integrated on-capillary tubular electrochemical detector for capillary electrophoresis systems has been fabricated based on sol-gel technique. It consists of a sol-gel carbon composite tubular electrode attached permanently onto the outlet of the separation capillary. The device greatly eases the setting up of capillary electrophoresis with electrochemical detection (CEEC) as it makes possible electrode/capillary alignment without the aid of a micromanipulator since this integrated unit can be simply immersed in the CE separation buffer in an ordinary three-electrode stationary cell. To improve analytical performance of the integrated unit, the external wall of the exit capillary was etched with HF after the polyimide coating of the capillary had been removed. Influences of the working electrode length and the wall thickness at the outlet of capillary on the separation efficiency and amperometric sensitivity were assessed and optimized. The practical applicability of this configuration is demonstrated with the detection of both catecholamines and carbohydrates. The advantages, namely, versatility, convenience, ease of operation, and low-cost, of the new design combined with an excellent performance lead to high stability and low detection limits. PMID- 11055696 TI - Effect of a variety of organic additives on retention and efficiency in micellar liquid chromatography AB - The effect of 21 organic additives (alkanols, alkane diols, dipolar aprotic solvents, alkanes) on the chromatographic behavior (retention, elution strength, efficiency) of probe solutes of widely differing hydrophobicity, such as benzene and 2-ethylanthraquinone, have been examined using a C18 stationary phase and sodium dodecyl sulfate (SDS) micellar mobile phases. The mobile-phase elution strength parallels the octanol-water partition coefficients of the additives or their ability to bind to the SDS micellar system, due to the increased solubility in the mobile phase and reduced affinity for the additive-modified surfactant coated stationary phase. The comparison of the elution strength of micellar mobile phases with that of a reference acetonitrile-water system indicates that the elution strength is lower for micellar systems and depends on the nature of the eluted solute. The displacement of the solute-micelle and solute-stationary phase binding equilibria is quantified for several probe solutes eluted with micellar mobile phases in the presence of 1-propanol, 1-butanol, 1-pentanol, and acetonitrile. A correlation was also observed between the number of theoretical plates and the hydrophobicity of the alcohol additives: the efficiency initially increased steeply and reached a plateau. Compared to benzene, a more hydrophobic additive was needed to attain the maximum efficiency for the more hydrophobic 2 ethylanthraquinone analyte. Dipolar aprotic solvents appear to be somewhat more effective in enhancing the efficiency than alcohols. The results are rationalized in terms of the ability of the organic additives to alter the composition, structure, dynamics, and properties of the micelles and the surfactant-coated stationary phase. PMID- 11055697 TI - Spectroscopic characterization of ethyl xanthate oxidation products and analysis by ion interaction chromatography AB - An ion interaction chromatographic separation method, coupled with UV spectroscopic detection, has been developed for the analysis of ethyl xanthate (O ethyl dithiocarbonate) and its oxidative decomposition products in mineral flotation systems. The effects of the ion-pairing agent (tetrabutylammonium ion), pH modifier (phosphoric acid), and organic modifier (acetonitrile) in the eluant upon the retention characteristics of the ethyl xanthate oxidation products have been determined. The optimized separation procedure has been successfully applied to the analysis of ethyl xanthate and its oxidation products in a nickel-iron sulfide mineral suspension containing a number of other anionic species, including cyanide complexes of nickel and iron, as well as sulfur-oxy anions. The ethyl xanthate oxidation products investigated in this study have been isolated as pure compounds and characterized by UV-visible, FT-IR, and NMR spectroscopies. The UV-visible and FT-IR spectroscopic properties of these species are discussed in terms of the chemical modifications of the thiocarbonate group. PMID- 11055698 TI - Column efficiency and separation of DNA fragments using slalom chromatography: hydrodynamic study and fractal considerations. AB - Novel equations (Guillaume Y. C.; et al. Anal. Chem. 2000, 72, 853) were developed to describe the large double-stranded DNA molecule retention in slalom chromatography (SC). These equations were applied for the first time to model both the "apparent selectivity" and the resolution between two eluted DNA fragments on a chromatogram. A study of the column efficiency corroborated the fact that slalom chromatography is not based on an adsorption or equilibrium phenomenon, but can be attributed to a hydrodynamic phenomenon. Using a combination of the dynamics of DNA fragment progression in the column and fractal considerations, it was shown that the apparent selectivity depends both on the DNA fragment sizes and mobile-phase flow rate and therefore a balance between two hydrodynamic regimes. A chromatographic response function was also used to obtain the most efficient separation conditions for a mixture of DNA fragments in a minimum analysis time. The chromatographic data confirmed that in SC the flow rate can increase or maintain the separation efficiency with an associated decrease in the analysis time. This constitutes an attractive outcome in relation to the classical chromatographic separation. PMID- 11055699 TI - Peak broadening in protein chromatography with monoliths at very fast separations. AB - Monoliths are stationary phases cast as a continuous medium which are interlaced by flow channels ramified with micropores. Pulse response experiments with bovine serum albumin as a model protein were applied for testing polymethacrylate-based monoliths, resulting in peak broadening that practically was not influenced by the chromatographic velocity. An empirical model was developed to describe peak broadening, allowing a term to account for the pore convection and a term for the pore diffusion. A diffusional distance lower than 10 nm was estimated. This corresponds to values observed with monodisperse 1-microm particles. Systematic investigations by changing the response time of the detector showed that the full potential of the monoliths could not be exploited, since the currently available chromatography systems are the limiting factor regarding the speed of data acquisition and virtual peak broadening by the infinite length of the detector. Inertia of the liquid and synchronization between liquid handling and electronic control introduced an additional disturbance. At the lowest possible response time, reliable peak data could be obtained up to a velocity of 35 cm/min. The pressure drop along the continuous bed was much smaller compared to a conventionally packed bed. Different flow patterns and significantly reduced eddy vortexes may be responsible for the high specific permeability. PMID- 11055700 TI - Identification of chlorinated dimethoxystilbene isomers and homologues in bleached paper products AB - We report here the identification of a novel class of compounds, the chlorinated dimethoxystilbenes (pinosylvins), in bleached paper products. Pinosylvins are naturally occurring compounds in wood that deter infections and predators. These compounds are being chlorinated during the pulp bleaching process. Two dichlorinated pinosylvins have previously been identified in bleached paper pulp, but we have identified other isomers and more highly chlorinated homologues in various bleached paper products. These compounds are present at concentrations on the order of hundreds of ppb. On the basis of the mass spectra of synthesized standards, we can distinguish the isomers in which the chlorines are located on the ethylene moiety from those in which the chlorines are located on the aromatic ring with the methoxy groups. In addition, we can predict the chlorine substitution patterns on both rings for these compounds using linear relationships between retention indexes and calculated dipole moments. The toxicity of these compounds is not known; however, isomers related to the pesticide methoxychlor are proestrogenic. PMID- 11055701 TI - In situ derivatization/solid-phase microextraction for the determination of haloacetic acids in water AB - An in situ derivatization solid-phase microextraction method has been developed for the determination of haloacetic acids (HAAs) in water. The analytical procedure involves derivatization of HAAs to their methyl esters with dimethyl sulfate, headspace sampling using solid-phase microextraction (SPME), and gas chromatography-ion trap mass spectrometry (GC/ITMS) determination. Parameters affecting both derivatization efficiency and head-space SPME procedure, such as the selection of the SPME coating, derivatization-extraction time and temperature, and ionic strength, were optimized. The commercially available Carboxen-poly(dimethylsiloxane) (CAR-PDMS) fiber appears to be the most suitable for the determination of HAAs. Moreover, the formation of HAA methyl esters was dramatically improved (up to 90-fold) by the addition of tetrabutylammonium hydrogen sulfate (4.7 micromol) to the sample as ion-pairing agent in the derivatization step. The precision of the in situ derivatization/HS-SPME/GC/ITMS method evaluated using an internal standard gave relative standard deviations (RSDs) between 6.3 and 11.4%. The method was linear over 2 orders of magnitude, and detection limits were compound-dependent, but ranged from 10 to 450 ng/L. The method was compared with the EPA method 552.2 for the analysis of HAAs in various water samples, and good agreement was obtained. Consequently, in situ derivatization/HS-SPME/GC/ITMS is proposed for the analysis of HAAs in water. PMID- 11055702 TI - On-line LC-MS analysis of urinary porphyrins. AB - A reversed-phase high-performance liquid chromatography-mass spectrometry (LC-MS) method is described for the separation and simultaneous analysis of porphyrins related to disorders of heme biosynthesis (uro-, heptacarboxylic, hexacarboxylic, pentacarboxylic, and coproporphyrins). The method involves initial porphyrin esterification and extraction from urine. Detection and quantification is performed from the extracts by separation with a Hypersil BDS column and on-line detection by MS through coupling with an atmospheric pressure chemical ionization interface. The porphyrin esters are detected as protonated molecules [M + H]+. Their mass spectra also exhibit an [M + Na]+ fragment of lower intensity. The analytical performance of this method is compared with those of LC with UV and fluorescence detection. LC-MS used in selective [M + H]+ ion monitoring provides the lowest detection and quantitation limits. In scan mode, this LC-MS method affords, without further isolation or concentration steps, the measurement of mass spectra of unknown compounds present in the urine of patients with altered porphyrin excretion. PMID- 11055703 TI - Isotopic analysis of uranium in tree bark by ICP mass spectrometry: a strategy for assessment of airborne contamination AB - Isotopic analysis of uranium in tree bark by ICP mass spectrometry is proposed as a new measurement strategy for monitoring airborne contamination and for discrimination of nuclear and nonnuclear emission sources. A quadrupole-based ICP mass spectrometer equipped with a microconcentric nebulizer and membrane desolvator was used to provide high-sensitivity measurement. The limit of detection for uranium (238U) was 0.004 ng L(-1). Measurement precision (235U/238U) was between 0.2 and 0.5% RSD for isotopic SRMs (U005 and U015; concentration, 1 microg L(-1)) and ranged from 0.4 to 3.1% RSD for tree bark extracts (U concentration, 0.03-0.08 microg L(-1)). Bark samples collected from the Peak District National Park in Derbyshire (U.K.) exhibited a natural 235U/238U isotope ratio value (0.0072) whereas samples from Sellafield, West Cumbria (U.K.) showed depletion in 235U (235U/238U = 0.0053-0.0064). PMID- 11055704 TI - Efficient preconcentration and separation of actinide elements from large soil and sediment samples AB - Large (approximately 10 g) soil and sediment samples are often required for analysis of low-level man-made actinides for monitoring environmental radioactivity. However, use of large environmental samples often results in significant chemical losses during chemical separations due to matrix interferences. We present a technique based on the ion-exchange resin Diphonix which selectively collects actinides and lanthanides into a common form, which then behaves uniformly and predictably during subsequent separation schemes. Diphonix is attractive for this purpose because it has an extremely high affinity for actinides and a low affinity for most common ions and is resistant to hydrofluoric acid. After being adsorbed onto Diphonix resin in a column mode at a pH approximately 1, actinides are completely eluted with 0.5 M 1 hydroxyethylidene-diphosphonic acid (HEDPA). After destruction of the HEDPA by ozonation or use of Fenton's reagent, Am, Pu, U, and Th are separated from each other and the remaining matrix by use of extraction chromatographic resins. We obtained high and consistent chemical recoveries (mean, 85%), as well as excellent chemical separations, for Am, Pu, U, and Th through the entire procedure for several 10-g soil and sediment samples. PMID- 11055705 TI - Real-time detection of organic compounds in liquid environments using polymer coated thickness shear mode quartz resonators AB - The selection of sensitive coatings is a critical task in the design and implementation of chemical sensors using coated thickness shear mode quartz crystal resonators (QCRs) for detection in liquid environments. This design or selection is performed through a study of the sorption process in terms of the partition coefficients of the analytes in the coatings. The partition coefficient, which is controlled by the chemical and physical properties of the coating materials, determines the inherent selectivity and sensitivity toward analyte molecules. The selection of the coatings is logically determined by the interactions between coating and target analyte molecules, but can also be made through a systematic variation of the coating's properties. The determination of the partition coefficients is only accurate if all contributions to the total measured frequency shifts, deltafs, of the coated QCR can be established. While mass loading is often assumed to be the dominant factor used in determining partition coefficients, viscoelastic effects may also contribute to deltafs. Both the effect of viscoelastic properties and the effect of mass loading on the sensor responses are investigated by using a network analyzer and oscillator circuit and by characterizing the total mechanical impedance of the loaded sensor. Different types of coatings including rubbery and glassy polymers are investigated, and the targeted analytes include classes of polar compounds (methanol), nonpolar compounds (toluene, xylenes), and chlorinated hydrocarbons (trichloroethylene, tetrachloroethylene, etc). It is seen that changes in viscoelastic properties due to analyte sorption may be significant enough to place the sensor in the nongravimetric regime. However, for most applications involving the detection of relatively low concentrations of organic compounds and the use of acoustically thin films, changes in the complex shear modulus of the coatings contribute less than 5% of the total shift in the series resonant frequency, depending on the coating. In that case, the measured deltafs and, hence, the calculated approximate classification and selection of the coatings for operation in a complex solution of water/analyte molecules. PMID- 11055706 TI - Plastic microfluidic devices modified with polyelectrolyte multilayers AB - Control of the polymer surface chemistry is a crucial aspect of development of plastic microfluidic devices. When commercially available plastic substrates are used to fabricate microchannels, differences in the EOF mobility from plastic to plastic can be very high. Therefore, we have used polyelectrolyte multilayers (PEMs) to alter the surface of microchannels fabricated in plastics. Optimal modification of the microchannel surfaces was obtained by coating the channels with alternating layers of poly(allylamine hydrochloride) and poly(styrene sulfonate). Polystyrene (PS) and poly(ethylene terephthalate) glycol (PETG) were chosen as substrate materials because of the significant differences in the polymer chemistries and in the EOF of channels fabricated in these two plastic materials. The efficacy of the surface modification has been evaluated using XPS and by measuring the EOF mobility. When microchannels prepared in both PS and PETG are modified with PEMs, they demonstrate very similar electroosmotic mobilities. The PEMs are easily fabricated and provide a means for controlling the flow direction and the electroosmotic mobility in the channels. The PEM coated microchannels have excellent wettability, allowing facile filling of the channels. In addition, the PEMs produce reproducible results and are robust enough to withstand long-term storage. PMID- 11055707 TI - Development of a time-resolved fluorometric detection system using diffusion enhanced energy transfer AB - A novel detection system using both emission energy transfer and time-resolved fluorometry (TRF) was developed, with a europium chelate as the energy donor and a novel fluorophore SNR1, excitable with long-wavelength light corresponding to europium emission, as the energy acceptor. When the donor and acceptor molecules were mixed in solution, energy transfer was observed without direct attachment of the donor and the acceptor, via a diffusion-enhanced energy-transfer mechanism. Thus, the acceptor emission can be detected as a long-lifetime fluorescence in TRF. When the fluorescence properties of the acceptor molecule are changed by interaction with an enzyme or other bioactive molecule, the change can be detected as a long-lived sensitized emission. If we develop or select suitable acceptor molecules, this simple and convenient system should be applicable to a wide variety of bioactive molecules. Since it is based on TRF, it can be used for high-resolution assay. PMID- 11055708 TI - Sulfonation of poly(N-vinylcarbazole) studied by combined time-of-flight secondary ion mass spectrometry and X-ray photoelectron spectroscopy AB - A series of sulfonated poly(N-vinylcarbazole) (PVK) samples have been systematically studied by time-of-flight secondary ion mass spectrometry (TOF SIMS) and X-ray photoelectron spectroscopy (XPS). Negative TOF-SIMS results provided unambiguous evidence that sulfonate groups are chemically attached to the carbazole moiety of PVK. The positive SIMS spectrum of PVK was, however, little affected by the sulfonation reaction. The degree of sulfonation was quantitatively determined by XPS. Therefore, the combination of TOF-SIMS and XPS is useful to follow the sulfonation reaction, both qualitatively and quantitatively. The SIMS intensities of some characteristic fragments are linearly related to the degree of sulfonation, suggesting that quantitative analysis is possible from TOF-SIMS data. PMID- 11055709 TI - Laser interference pattern ablation of a carbon fiber microelectrode: biosensor signal enhancement after enzyme attachment. AB - Fluorescence microscopy was used to visualize the accumulated fluorescent product of the enzyme alkaline phosphatase to indicate where active covalently bound enzyme remained on the surface after application of a Nd: YAG laser interference pattern to a surface that was first globally derivatized with the covalently bound enzyme. The electrochemical kinetics of the same carbon fiber surface were examined through the electrogenerated chemiluminescence of Ru(bpy)(3)2+ to determine that electron-transfer sites were indeed segregated from the enzyme binding sites. The enzyme-derivatized areas are determined to be separate and distinct from the areas of enhanced electron transfer. Two other enzymes, glucose oxidase and malic dehydrogenase, were then covalently bound to carbon fiber microelectrode surfaces in order to verify the change in detection limit of their respective cofactors, NADH or H2O2, under a variety of surface conditions. The S/N of an enzyme-modified electrode after laser interference pattern photoablation and electrocatalytic treatment is improved by more than 1 order of magnitude over that observed at an electrode that is globally enzyme modified. PMID- 11055710 TI - Microscopic measurement of pH with iridium oxide microelectrodes AB - Microscopic pH electrodes were produced by deposition of hydrous iridium oxide onto carbon fiber microelectrodes. The electrodes exhibit two linear regions of potentiometric response between pH 2-6 and pH 6-12. The electrodes respond to pH changes within 50 ms, and an equilibrium value is reached within 30 s. By using these electrodes as probes in the scanning electrochemical microscope, dynamic pH changes occurring at or near a surface can be measured and pH maps of the surface can be generated. Vertical pH profiles and images of pH were obtained at substrates where electrochemical (oxidation and reduction of H2O2, hydrogen evolution) or enzymatic (glucose oxidase) reactions involving proton transfers occur. PMID- 11055711 TI - Chromatographic detection of nitroaromatic and nitramine compounds by electrochemical reduction combined with photoluminescence following electron transfer. AB - The oxidizing agent tris(bipyridyl)ruthenium(III), or Ru-(bpy)(3)3+, is used as a postcolumn reagent for the detection of nitroaromatic and nitramine explosive compounds. After separation, the explosives are reduced electrochemically to oxidizable products such as hydroxlamines and nitrosamines, and these products react readily with Ru-(bpy)(3)3+ and Ru(bpy)(3)2+. The photoluminescence from the latter is used for detection. A porous carbon electrode was used for on-line analyte reduction following chromatography. Another porous carbon electrode was used to generate the nonluminescent Ru(bpy)(3)3+ from Ru(bpy)(3)3+ on-line at high efficiency. The two streams were combined, and the Ru(bpy)(3)2+ produced by oxidation of the reduced analytes was detected by laser illumination and light detection. Reductive hydrodynamic voltammograms of nitrobenzene, 2,4,6 trinitrotoluene, and hexahydro-1,3,5-trinitro-1,3,5-triazine indicated that a potential of - 1500 mV vs Ag/AgCl was sufficient to achieve a maximum signal from the reduced analytes. HPLC with a water/acetonitrile gradient on a C-18 reversed phase column was then used to determine these three compounds plus the four additional examples, 1,3,5,7-tetrazocine, 2,4-dinitrotoluene; 2,6-dinitrotoluene, and 4-nitrotoluene. For both hydrodynamic voltammetry and HPLC detection, the photoluminescence following electron-transfer signal was calibrated using the one electron standards ferrocene and ferrocenecarboxylic acid. Detection limits were in the low-nanomolar range for 20-microL injections of nonpreconcentrated nitro compounds. PMID- 11055712 TI - A single calibration graph for the direct determination of ascorbic and dehydroascorbic acids by electrogenerated luminescence based on Ru(bpy)(3)2+ in aqueous solution AB - Ascorbic (H2A) and dehydroascorbic (DA) acids were for the first time directly determined in a single chromatographic run by means of the tris(2,2' bipyridine)ruthenium(II) (Ru(bpy)(3)2+) based electrogenerated chemiluminescence (ECL) detection. For the first time, it was demonstrated that DA, a nonelectroactive compound, is ECL active and is responsible for the ECL behavior of H2A. This fact, together with the lack of a DA standard, suggested the use of a calibration graph obtained for H2A, for determining both analytes. The proven ECL activity of DA, together with literature data relative to the standard redox potentials of the different species coming from H2A, led to a reconsideration of the proposed ECL reaction mechanism for H2A. The role of the OH- ion in the reaction mechanism of the two analytes appeared to be crucial. H2A and DA could be separated by a suitable C18-reversed-phase HPLC column using an aqueous 30 mM H3PO4 solution as the mobile phase. The optimal ECL response was achieved by polarizing the working electrode at 1.150 Vvs SCE (standard calomel electrode) (oxidation diffusion limiting potential for both H2A and Ru(bpy)(3)2+). The Ru(bpy)(3)2+ solution, at pH 10 for carbonate buffer, was mixed to the eluent solution in a postcolumn system, obtaining, still at pH 10, the final 0.25 mM Ru(bpy)(3)2+ concentration. The detection limit found for the two analytes was 1 x 10(-7) M. The method was successfully applied to the determination of the analytes in a commercially available orange fruit juice. PMID- 11055713 TI - Scanning electrochemical microscopy. 40. Voltammetric ion-selective micropipet electrodes for probing ion transfer at bilayer lipid membranes. AB - Voltammetric ion-selective micropipet electrodes for use in scanning electrochemical microscopy (SECM) for detection of potassium ion were fabricated. These used pulled borosilicate capillaries with tip orifice radii of 0.7-20 microm with silanized inner walls filled with a solution of 10 mM valinomycin and 10 mM ETH 500 in dichloroethane. The electrodes were characterized by determining the steady-state tip current for K+ concentrations of 0.05-0.3 mM. The tips were used in the SECM feedback and generation-collection modes to study K+ transfer through gramicidin channels in a horizontal bilayer lipid membrane (glycerol monooleate). PMID- 11055714 TI - Ferricyanide reduction by Escherichia coli: kinetics, mechanism, and application to the optimization of recombinant fermentations. AB - Ferricyanide reduction was studied by flow injection analysis (FIA) and chronoamperometry (CA) using two host strains and one recombinant strain of E. coli. Samples taken from batch cultures of E. coli JM105 and HB101 showed maximal specific ferricyanide reduction rates in the late exponential phase of growth, with values (micromol/min x g) of 24 (FIA) and 17 (CA) for JM105, and 36 (FIA) for HB101, when shake-flask cultures were sampled, and 70 for HB101, when a chemostat was used to control pH and dissolved oxygen concentration throughout the cultivation. Remarkably higher ferricyanide reduction rates were obtained with HB101 cells cultivated continuously at very slow growth rate, when chilled, resuspended cell samples were incubated for 5 min in solutions containing 10 mM succinate or formate. These compounds are substrates for primary, membrane-bound dehydrogenases that transfer electrons via ubiquinone to the cytochrome oxidase complexes. Apparent Michaelis-Menten kinetics were observed with respect to ferricyanide concentration when 10 mM succinate was included in the assay buffer; apparent Km values of 10.1+/-0.6 mM and 14.4+/-1.2 mM ferricyanide were obtained for exponential- and stationary-phase E. coli JM105, respectively. Cyanide inhibition studies show that ferricyanide is reduced mainly by cytochrome o oxidase in exponentially growing cells. The large difference in ferricyanide reduction rates observed in the absence and presence of succinate and formate were used to signal stationary-phase entry 5 h after induction of recombinant human Cu/Zn superoxide dismutase expression in a batch fermentation of E. coli HMS174(DE3)(pET3ahSOD). This new method can be used as an adjunct to the quantitation of medium components for the optimization of recombinant fermentations. PMID- 11055715 TI - Rapid antibiotic susceptibility testing via electrochemical measurement of ferricyanide reduction by Escherichia coli and Clostridium sporogenes. AB - Electrochemical measurement of respiratory chain activity allows rapid and reliable screening for antibiotic susceptibility in microorganisms. Chronoamperometry and chronocoulometry of suspensions of aerobically cultivated E. coli combined with the non-native oxidant potassium hexacyanoferrate(III) (ferricyanide) yield signals for reoxidation of the reduction product ferrocyanide that are much smaller if the E. coli has been incubated briefly with an effective antibiotic compound. Chronocoulometric results, obtained following 20-min incubation with antibiotic and 2-min measurement in assay buffer containing 50 mM ferricyanide and 10 mM succinate, at +0.50 V vs Ag/AgCl at a Pt working electrode, were compared with traditional disk diffusion susceptibility testing, which requires overnight incubation on agar plates; the results show significantly lower accumulation of ferrocyanide in all cases in which growth inhibition was observed in the disk diffusion assay. A range of antibiotic compounds (13) were examined that possess different mechanisms of action. Quantitative determination of IC50 values for penicillin G and chloramphenicol yielded values that were 100-fold higher than those obtained by standard turbidity methods after 10-h incubation; this is likely a result of the very brief (10 min) exposure time to the antibiotics. Addition of 5 microM 2,6 dichlorophenolindophenol, a hydrophobic electron-transfer mediator, to the assay mixture allowed susceptibility testing of a Gram-positive obligate anaerobe, Clostridium sporogenes. This rapid new assay will facilitate clinical susceptibility testing, allowing appropriate treatment virtually as soon as a clinical isolate can be obtained. PMID- 11055716 TI - Modeling of divalent/monovalent ion selectivity of ion-exchanger-based solvent polymeric membranes doped with coexchanger AB - It is shown that the addition of a coexchanger does not always improve the selectivity of an ion-selective electrode. As a model case, the selectivity toward divalent primary ions I2+ in the presence of monovalent interference J+ is numerically simulated for membranes based on ion-exchanger R- and doped with a coexchanger (ionic additive) S+. It is, for the first time, demonstrated that the addition of a coexchanger, together with a high mobility of ion-exchanger sites, may be either beneficial or harmful to selectivity. Accordingly, two distinctive representatives of systems, i.e., membrane/solution are discussed. The theory is illustrated with experimental data obtained from PVC membranes based on calcium bis(2ethylhexyl)-phosphate plasticized with tris(2ethylhexyl)phosphate and doped with tetradecylammonium. Electrodes with these membranes exhibit both types of selectivity behavior. Interference from lithium with a calcium response may be sufficiently decreased by the addition of tetradecylammonium, while the interference from potassium and cesium was increased in the doped membranes. PMID- 11055717 TI - Isolation of n-decyl-alpha(1-->6) isomaltoside from a technical APG mixture and its identification by the parallel use of LC-MS and NMR spectroscopy AB - The identification of n-decyl alpha(1-->6)isomaltoside as a main component of technical alkyl polyglucoside (APG) mixtures by the parallel use of liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) spectroscopy is described. Following enrichment on a styrene-divinylbenzene-based solid-phase extraction material, unknown components were separated by reversed phase liquid chromatography (LC). Chemical characterization was achieved by both mass spectrometry and NMR spectroscopy. It is demonstrated that the combination of LC-MS with various NMR techniques is very suitable for stereochemical assignment of unknown components in technical APG mixtures. PMID- 11055718 TI - Room temperature sonolysis-based advanced oxidation process for degradation of organomercurials: application to determination of inorganic and total mercury in waters by flow injection-cold vapor atomic absorption spectrometry AB - A new oxidation method based on room-temperature ultrasonic irradiation (sonolysis) is proposed for conversion of organomercurials into inorganic mercury and subsequent determination by flow injection-cold vapor atomic absorption spectrometry. This advanced oxidation process eliminates the need for chemical oxidants, high temperature, and pressure for degradation of organomercurials so that total mercury can be determined with sodium tetrahydroborate(III) or tin(II) chloride as reducing agents. Complete oxidations can be accomplished within 3 min, using a 40% sonication amplitude (100 W nominal power) provided by a probe ultrasonic device (20 kHz frequency) and a 1 mol L(-1) HCl liquid medium. The presence of HCl was seen to be necessary for fast oxidation of organomercurials, in contrast to other chemical oxidants such as H2O2 or HNO3 which yielded incomplete oxidation. Further advantages of the proposed method over existing methods which are currently employed for oxidation prior to total Hg determination are the removal of hazardous wastes and the decreased risk of Hg losses by volatilization. Oxidation kinetics indicated a pseudofirst-order reaction with apparent rate constants (k) of 3.2 x 10(-2) and 1.6 x 10(-2) s(-1) for methylmercury and phenylmercury, respectively. Oxidation experiments in the presence of foreign substances acting as OH radical scavengers showed a tolerance at least up to a concentration of 1000 mg L(-1). Likewise, model wastewaters with chemical oxygen demand of up to 1000 mg L(-1) could be processed without diminishing the oxidation efficiency. The method was applied to determination of inorganic and total mercury in simulated wastewaters and spiked environmental waters in combination with selective reduction. PMID- 11055719 TI - Temperature-insensitive near-infrared method for determination of protein concentration during protein crystal growth. AB - A temperature-insensitive method for measuring protein concentration in aqueous solutions using near-infrared spectroscopy is described. The method, which is based on identification of the net analyte signal of single-beam spectra, can be calibrated using a single protein absorbance measurement and is thus well suited for crystallization monitoring where the quantity of protein is limited. The method is applied to measurements of glucose-isomerase concentration in a sodium phosphate buffer that is actively varied over the temperature range of 4-24 degrees C. The standard error of prediction using the optimized spectral range of 4670-4595 cm(-1) is 0.12 mg/mL with no systematic trend in the residuals with solution temperature. The method is also applied to previously collected spectra of hen egg-white lysozyme and yields a standard error of prediction of 0.14 mg/mL. Spectra sampled at discrete wavelengths can also be used for calibration and prediction with performance comparable to that obtained with spectral bands. A set of four wavelengths are identified that can be used to predict concentrations of both proteins with a standard error less than 0.14 mg/mL. PMID- 11055720 TI - Monitoring temperature changes in capillary electrophoresis with nanoliter-volume NMR thermometry. AB - Nanoliter-volume proton nuclear magnetic resonance (NMR) spectroscopy is used to monitor the electrolyte temperature during capillary electrophoresis (CE). By measuring the shift in the proton resonance frequency of the water signal, the intracapillary temperature can be recorded noninvasively with subsecond temporal resolution and spatial resolution on the order of 1 mm. Thermal changes of more than 65 degrees C are observed under both equilibrium and nonequilibrium conditions for typical CE separation conditions. Several capillary and buffer combinations are examined with external cooling by both liquid and air convection. Additionally, NMR thermometry allows nonequilibrium temperatures in analyte bands to be monitored during a separation. As one example, a plug of 1 mM NaCl is injected into a capillary filled with 50 mM borate buffer. Upon reaching the NMR detector, the temperature in the NaCl band is more than 20 degrees C higher than the temperature in the surrounding buffer. Such observations have direct applicability to a variety of studies, including experiments which utilize sample stacking and isotachophoresis. PMID- 11055721 TI - A liquid chromatography-mass spectrometry method for the quantification of bioavailability and bioconversion of beta-carotene to retinol in humans. AB - A method based on high-performance liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (APCI LC-MS) was developed for the quantification of the bioavailability of retinyl palmitate and beta-carotene and the bioconversion of beta-carotene to retinol in humans. Following oral administration of [8,9,10,11,12,13,14,15,19,20-13C10]-retinyl palmitate and [12,13,14,15,20,12',13',14',15',20'-13C10]-beta-carotene at physiological doses to children between 8 and 11 years of age, blood samples were drawn and serum was prepared. Retinol and beta-carotene were extracted from 0.2- and 1.0-mL serum samples, respectively, and analyzed using reversed-phase HPLC with a C30 column interfaced to an APCI mass spectrometer. Unlike other LC-MS assays for carotenoids, no additional purification steps were necessary, nor was any derivatization of retinol or beta-carotene required. APCI LC-MS showed a linear detector response for beta-carotene over 4 orders of magnitude. Using selected ion monitoring to record the elution profile of protonated circulating beta carotene at m/z 537 and [13C10]-beta-carotene at m/z 547, the limit of detection was determined to be 0.5 pmol injected on-column. To assess the ratio of labeled to unlabeled retinol, selected ion monitoring was carried out at m/z 269, 274, and 279. These abundant fragment ions corresponded to the loss of water from the protonated molecule of circulating retinol, [13C5]-retinol (metabolically formed from orally administered [13C10]-beta-carotene), and [13C10]-retinol (formed by hydrolysis of [13C10]-retinyl palmitate). The ratios of labeled to unlabeled retinol and the ratio of labeled to unlabeled beta-carotene were calculated. Combined with standard HPLC measurement of beta-carotene and retinol concentration and a mathematical model, these results showed that this simple LC MS method can be used to quantify beta-carotene bioavailability and its bioconversion to retinol at physiologically relevant doses. PMID- 11055722 TI - Evaluation of neural network models with generalized sensitivity analysis AB - A sensitivity analysis method for discovering characteristic features of the input data using neural network classification models has been devised. The sensitivity is the gradient of the neural network model response function, and because neural network models are nonlinear, the gradient depends on the point where it is evaluated. Two criteria are used for measuring the sensitivity. The first criterion calculates the sensitivity or gradient of the neural network output with respect to the average of the objects that comprise each class. The second criterion measures the average sensitivity of the class objects. The sensitivity analysis was applied to temperature-constrained cascade correlation network models and evaluated with sets of synthetic data and experimental mobility spectra. The neural network models were built using temperature constrained cascade correlation networks (TCCCNs). A weight constraint was devised for the output units of the network models. This method implements weight decay with conjugate gradient training and yields more sensitive neural network models. Temperature-constrained hidden units furnish more sensitive network models than networks without constraints. By comparing the sensitivities of the class mean input and the mean sensitivity for all the inputs of a class, the individual input variables may be assessed for linearity. If these two sensitivities for an input variable differ by a constant factor, then that variable is modeled by a simple linear relationship. If the two sensitivities vary by a nonconstant scale factor, then the variable is modeled by higher order functions in the network. The sensitivity method was used to diagnose errors in the training data, and the test for linearity indicated a TCCCN architecture that had better predictability. PMID- 11055723 TI - Improved ion transmission from atmospheric pressure to high vacuum using a multicapillary inlet and electrodynamic ion funnel interface AB - A heated multicapillary inlet and ion funnel interface was developed to couple an electrospray ionization (ESI) source to a high-vacuum stage for obtaining improved sensitivity in mass spectrometric applications. The inlet was constructed from an array of seven thin-wall stainless steel tubes soldered into a central hole of a cylindrical heating block. An electrodynamic ion funnel was used in the interface region to more effectively capture, focus, and transmit ions from the multicapillary inlet. The interface of seven capillary inlets with the ion funnel showed more than 7 times higher transmission efficiency compared to that of a single capillary inlet with the ion funnel and a 23-fold greater transmission efficiency than could be obtained using the standard orifice-skimmer interface of a triple-quadrupole MS. The multiple-capillary inlet and ion funnel interface showed an overall 10% ion transmission efficiency and approximately 3 4% overall detection efficiency of ions from solution based (i.e., prior to electrospray). The improved performance was achieved under conditions where ESI operation is robust and results in a significant increase in dynamic range. PMID- 11055724 TI - Separation and identification of twelve catechins in tea using liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry. AB - A method has been developed for the direct microscale determination of 12 catechins in green and black tea infusions. The method is based on liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry (LC/APCI-MS). Standard catechin mixtures and tea infusions were analyzed by LC/APCI-MS with detection of protonated molecular ions and characteristic fragment ions for each compound. The identities of eight major catechins and caffeine in tea were established based on LC retention times and simultaneously recorded mass spectra. In addition, monitoring of the catechin-specific retro Diels-Alder fragment ion at m/z 139 throughout the chromatogram provided a unique fingerprint for catechin content in the samples that led to the identification of four minor chemically modified catechin derivatives in the infusions. This report is the first to describe the comprehensive determination of all 12 reported catechins in a single analysis. The utility of LC/APCI-MS for providing routine separation and identification of catechins at femtomole to low-picomole levels without extraction or sample pretreatment, and its potential as a standard analytical tool for the determination of polyphenols in natural products and biological fluids, are discussed. PMID- 11055725 TI - Negative ion mass spectrometry of sialylated carbohydrates: discrimination of N acetylneuraminic acid linkages by MALDI-TOF and ESI-TOF mass spectrometry. AB - Negative ion MALDI and electrospray fragmentation spectra were recorded from 12 sialylated carbohydrates ranging from trisaccharides to biantennary N-linked glycans. D-Arabinosazone was found to be the most satisfactory MALDI matrix for these compounds. Fragmentation mechanisms were investigated with the aid of several synthesized analogues of the sugars labeled with 13C and 2H. The substitution position of the sialic acid (alpha2-->3 or alpha2-->6) was found to have a dramatic effect on the overall fragmentation pattern of these compounds, and several features of the spectra were identified that allowed the substitution pattern to be determined. In particular, the appearance of an ion at m/z 306 appeared to be diagnostic of the presence of an alpha2-->6-linked sialic acid. Selection and further fragmentation of the in-source (conevoltage) fragment ion corresponding to the trisaccharide Neu5Acalpha2-->3(or 6)Galbeta1-->4GlcNAc from larger, N-linked glycans, ionized by electrospray, gave fragmentation patterns identical to those of the reference trisaccharides, thus providing a method for confirming the sialic acid linkage. PMID- 11055726 TI - Direct determination of soil surface-bound polycyclic aromatic hydrocarbons in petroleum-contaminated soils by real-time aerosol mass spectrometry AB - Soil surface-bound polycyclic aromatic hydrocarbons (PAHs) were identified by use of Real-Time Aerosol Mass Spectrometry (RTAMS) in two NIST standard research material (SRM) soils (Montana SRM 2710 and Peruvian SRM 4355) each contaminated separately with three common petroleum hydrocarbons (diesel fuel, gasoline, and kerosene). The described contaminated soil analysis required no sample preparation. Direct laser desorption/ionization mass spectrometric analysis of individual soil particles contaminated with each of the petroleum hydrocarbons at three different contamination levels (0.8, 8, and 80 ppth (wt/wt)) yielded detectable PAH cation distributions that ranged from m/z 128 to 234, depending on the fuel contaminant. The same analysis performed on uncontaminated SRM soils revealed very little (Peruvian) to no (Montana) detectable PAH species. Size analysis showed that most of the individual soil particles analyzed were between 1 and 5 microm in diameter. Tandem mass spectrometry (MS/MS) experiments identified alkyl-substituted two- and three-ringed PAHs in all three petroleum hydrocarbon contaminated soils. However, due to similarities in fragmentation patterns, MS/MS analysis of higher MW species (m/z > 200) was unable to distinguish between the possibility of highly alkyl-substituted three-ringed PAHs and hydrogenated four-ringed PAHs. The described technique offers the direct, rapid determination and characterization of surface-bound PAHs in petroleum contaminated soils at part-per-million levels without prior extraction, separation, or other sample preparation methods. PMID- 11055727 TI - Correction of species transformations in the analysis of Cr(VI) in solid environmental samples using speciated isotope dilution mass spectrometry AB - Speciation of Cr(VI) in solid environmental samples is challenging because of the transformations between Cr(VI) and Cr(III). EPA method 3060A completely extracts Cr(VI) in a hot alkaline solution and preserves the solublized Cr(VI). This procedure, however, can oxidize Cr(III) in some chemical forms. On the other hand, the reverse transformation may occur during neutralization and acidification following the extraction step. We developed a method that is capable of monitoring and correcting for such bidirectional species transformations to determine Cr(VI) in solid samples. In this method, we spike a sample with a 53Cr(VI) spike (enriched in 53Cr) and a isoCr(III) spike (enriched in 50Cr). The large quantity of isoCr(III) in an easily oxidizable form competes with sample Cr(III) in the oxidization, reducing the method-induced oxidation of sample Cr(III). This method also corrects for the reduction of Cr(VI). The theory is presented and is evaluated experimentally. The analysis of chromite ore processing residue, fly ash, and standard reference material SRM 1645 showed that the oxidation of sample Cr(III) could cause positive biases as high as 163% if no correction is performed. PMID- 11055728 TI - High-pressure ion source combined with an in-axis ion trap mass spectrometer. 1. Instrumentation and applications AB - A new combination of a dual EI/CI ion source with a quadrupole ion trap mass spectrometer has been realized in order to efficiently produce negative ions in the reaction cell. Analysis of volatile compounds was performed under negative ion chemical ionization (NICI) during a reaction period where selected reactant negative ions, previously produced in the external ion source, were allowed to interact with molecules, introduced by hyphenated techniques such as gas chromatography. The O2*-, CH3O-, and Cl- reactant ions were used in this study to ensure specific ion/molecule interactions such as proton transfer, nucleophilic displacement, or charge exchange processes, respectively leading to even-electron species, i.e., deprotonated [M - H]- molecules, diagnostic [M - R]- ions, or odd electron M*- molecular species. The reaction orientation depends on the thermochemistry of reactions within kinetic controls. First analytical results are presented here for the trace-level detection of several contaminants under NICI/Cl- conditions. Phosphorus-containing compounds (malathion, ethyl parathion, and methyl parathion as representative for pesticides) and nitro-containing compounds (2,4,6-trinitrotoluene for explosive material) have been chosen in order to explore the analytical ability of this promising instrumental coupling. PMID- 11055729 TI - High-pressure ion source combined with an in-axis ion trap mass spectrometer. 2. Application of selective low-pressure negative ion chemical ionization AB - Negative ion chemical ionization was carried out using a quadrupole ion trap mass spectrometer with selected reactant negative ions, primarily injected from a homemade dual EI/CI external ion source. Hence, selective ion/molecule reactions were provided according to the reaction time, which induce a greater control over bimolecular ionization mechanisms than in conventional a high-pressure ion source combined with beam instruments, where several competitive ionization processes take place mainly due to source conditions (e.g., temperature, pressure, and repeller). By selecting the reactant ions, ion/molecule reactions were specifically produced (i.e., charge exchange, proton transfer, nucleophilic substitution, and/or alpha-beta elimination) with several organic target compounds. Gas-phase reactivity of phosphorus- and nitrogen-containing compounds (such as phosphonates as representative for chemical warfare agents and phosphorothionates, phosphorodithionates, and triazines for pesticides) as well as dinitro aromatic compounds (for pesticides) has been explored, in the present work, to ensure further unambiguous detection. PMID- 11055730 TI - The use of deuterium oxide as a mobile phase for structural elucidation by HPLC/UV/ESI/MS AB - The use of deuterium oxide as a mobile phase in the routine analysis of pharmaceutical compounds was investigated. The deuterium exchange of labile hydrogen atoms aids in structural confirmation and elucidation of unknown impurities and degradation products. Although deuterium oxide as a mobile phase does in some cases change the retention times, the changes in retention times do not interfere with the analysis. A study of the high-performance liquid chromatography system shows that equilibration times for the deuterium-containing mobile phases are similar to equilibration times with changes of other mobile phases. The use of this technique in the analysis of pharmaceutical compounds and other small molecules is presented. PMID- 11055731 TI - Cylindrical ion trap array with mass selection by variation in trap dimensions AB - A mass spectrometer array is described in which each array element is a cylindrical ion trap (CIT) within which an approximately quadrupolar, time varying, field is established. The individual traps are of different sizes, so that when the array is operated with a fixed rf potential, ions of different masses (or mass ranges) are stored in each trap. By choosing the dimensions of each CIT element in the array, a multiple ion monitoring experiment can be performed. For example, in a two-element array with elements having internal radii of 5 and 4 mm, the smaller trap selects for m/z 91 and the larger for m/z 57, corresponding to characteristic aromatic and aliphatic hydrocarbon ions. Ion storage using both rf/dc (apex) isolation and the stored waveform inverse Fourier transform method is demonstrated.The array reduces the complexity of the electronics needed to operate the ion trap, which should make it suitable for use in a miniature mass spectrometer system. PMID- 11055732 TI - Characterization of polyphosphates by electrospray mass spectrometry AB - Electrospray ionization mass spectrometry (ESI-MS) was applied for the characterization of inorganic polyphosphates [orthophosphate, pyrophosphate, tripolyphosphate, trimetaphosphate, and tetrapolyphosphatel. The high selectivity of ESI-MS allows the detection of different polyphosphate species without preseparation by ion chromatography or capillary electrophoresis. Furthermore, ESI-MS does not require the incorporation of UV-absorbing chromophores into the analytical method for the detection of phosphates, unlike conventional UV chromatographic methods. Limits of detection by ESI-MS were estimated to range from approximately 1 to 10 ng/mL. The quantification of polyphosphate samples as single-component and multicomponent mixtures was investigated. Linear signal response for single-component samples ranged from the limit of detection to approximately 10 microg/mL Quantification of polyphosphate in streamwater is demonstrated using the standard addition method. The effect of multi polyphosphate components and salts on signal response was also studied. For concentrations less than 2.0 microg/mL, signal response from a tetrapolyphosphate sample was comparable to those obtained from tetrapolyphosphate-tripolyphosphate mixtures. Signal response obtained from tetrapolyphosphate in the presence of tripolyphosphate or NH4NO3 at higher concentrations (approximately 50 microg/mL and 35 microg/mL, respectively) was significantly lower relative to single component standards (approximately 40%-70%). PMID- 11055733 TI - Analysis of the degradation of oligonucleotide strands during the freezing/thawing processes using MALDI-MS. AB - Synthetic oligonucleotide strands ranging from 5 to 25 units in length are commonly used as standards, probes, and templates in various bioanalytical applications. Until recently, their preparation, storage, and handling were regarded as unimportant, but this work provides valuable information to the contrary. The systematic degradation of oligonucleotide strands during sample preparation is investigated by repeatedly freezing/thawing short strands followed by matrix-assisted laser desorption ionization mass spectrometric (MALDI-MS) analysis. It is shown here that the longevity of an oligonucleotide strand is dependent on several factors including base composition, solution concentrations, and strand length as well as thawing conditions. Several trends in strand robustness were established. Our studies reveal that the robustness of strands is base-dependent: T-mer > A-mer > C-mer > G-mer. Likewise, an increase in the length of the strands increases the tendency of a sample to degrade. Another observation included that samples of mixed bases degrade according to structural conformations. All of these observations are attributed to the fact that the samples undergo degradation during sample/solvent isolation during freezing. PMID- 11055734 TI - MALDI TOF mass spectrometry for the characterization of phosphorus-containing dendrimers. Scope and limitations AB - Neutral phosphorus-containing dendrimers with aldehyde groups at the periphery have been analyzed using matrix-assisted laser desorption ionization time-of flight mass spectrometry (MALDI-TOFMS) up to generation four. Although the expected quasi-molecular ion is generally observed, the mass spectral pattern, presence of fragments and adducts related to the original skeleton, is highly relevant to the sample preparation (nature of the matrix: 2-5-dihydroxybenzoic acid (2.5-DHB), 1,8-dihydroxy-9[10H]-anthracenone (dithranol), 6-azathiothymine, 2,4,6-trihydroxyacetophenon, 7-hydroxycoumarin or 2-anthramine, and addition of alkali metal salts). The dithranol matrix with addition of LiI offers milder conditions; however, abundant fragments are still observed for the higher generation dendrimers. Investigation of these effects in connection with SEC, NMR, and MALDI-TOFMS studies of UV preirradiated dendrimers allows the assumption to be made that fragmentation occurs in MALDI due to the relatively strong absorption of the dendrimers at 337 nm. Fragmentations and formation of adducts involve nitrogen-nitrogen bond cleavage, imine metathesis, and reaction of aldehyde groups with internal imino groups. PMID- 11055735 TI - Characterization of synthetic polymers by MALDI-TOF/MS: investigation into new methods of sample target preparation and consequence on mass spectrum finger print AB - Characterization of synthetic polymers by Matrix assisted laser desorption (MALDI) is limited by the solubility of different oligomers in a suitable solvent, and the fingerprint of the mass spectrum is affected by the properties of solvents employed (eg., pH, secondary solvents, evaporation) during sample target preparation. If solvents are not used during sample target preparation, then solvent properties should not play an important role in determining the quality of the MALDI mass spectrum. We report here two solventless approaches for sample target preparation. It was observed that Poly(ethylene glycol) 6000 (PEG) showed the same molecular mass distribution in different modes of sample target preparation. Fluorinated polymer used in these studies was affected by sample target preparation protocol and by target surface. Pyrolysis of PEG oligomers was observed in all the methods of sample target preparation. The desorbed high mass neutral oligomers fragment to give small oligomers which are then cationized by the desolvation of the cationized matrix clusters. Moreover, the origin of the matrix clusters (i.e., formed in the condensed phase or in the gas phase) determines the relative intensities of PEG oligomers cationized by sodium or potassium. PMID- 11055736 TI - Determination of condensed tannin monomers in environmental samples by capillary gas chromatography of acid depolymerization extracts AB - A method for molecular-level quantification of condensed tannin is described that uses acid depolymerization and carbocation capture by phloroglucinol. Resulting monomers and phloroglucinol adducts are trimethylsilyl derivatized, separated by capillary gas chromatography, detected by flame ionization, and quantified relative to standards. Optimal depolymerization conditions were determined for acid strength, phloroglucinol concentration, time, and temperature. The method gives reproducible results in leaf litter that are linear over 2 orders of magnitude with detection limits down to approximately 100 ng condensed tannin. In addition to tannin, triterpenoids were also identified and can be quantified with this method. Analyses of soils and sediments indicate that mineral interactions with condensed tannin are important both analytically and environmentally. PMID- 11055737 TI - Application of multishot acquisition in Fourier transform mass spectrometry. AB - A new method of ion injection and trapping is discussed wherein ions are accumulated over several laser shots in the FT-ICR cell prior to detection. This allows accumulation of ion signal without accumulating noise so that the signal/noise ratio is much improved provided that the "space-charge" limit of the total number of ions in the cell is not exceeded. "In-cell" ion accumulation allows selected ion accumulation by simply sweeping unwanted ions out of the cell prior to subsequent ion trapping events and also allows shifted ion accumulations to correct for time-of-flight distortions in the ion abundance distributions. PMID- 11055738 TI - Measurement of dilute 29Si species in solution using a large volume coil and DEFT NMR AB - A large-sample-volume nuclear magnetic resonance (NMR) spectroscopy probehead has been developed for the detection and characterization of low concentrations of 29Si species in aqueous solution. The approach described entails the use of a large-diameter radio frequency solenoid coil that permits substantially larger sample volumes to be investigated at moderate magnetic field strengths, compared with conventional NMR probehead configurations. In addition, difficulties presented by long 29Si T1 relaxation times have been circumvented by using the DEFT NMR pulse sequence, which permits more rapid signal averaging. Through a combination of these hardware and methodological improvements, high-resolution 29Si NMR spectra have been obtained at 4.2 T (29Si resonance frequency = 36.8 MHz) for an 800 microM solution of 96% 29Si-enriched silicic acid, H4SiO4 (pH approximately 8), with a signal-to-noise ratio of 16 and a line width of 31 Hz after 3 h of total measurement time. PMID- 11055739 TI - Phenyl- and butyltin analysis in small biological samples by cold methanolic digestion and GC/MS. AB - A very efficient technique for the analysis of six butyl- and phenyltin compounds in biota samples has been developed. No special equipment is needed for sample preparation, which is based on cold methanolic digestion with subsequent aqueous ethylation and liquid-liquid extraction. For samples of only 40 mg of biological materials, method detection limits ranging from 4 to 52 ng/g were achieved using gas chromatography/mass spectrometry. Relative recoveries for the individual butyl- and phenyltins, referring to perdeuterated organotin analogues as internal standards, ranged from 96 to 107%. Organotin concentrations in insect larvae (Chironomus riparius) and a reference mussel tissue (CRM 477) were determined with excellent precision (RSD <5%), and the measured butyltins in CRM 477 were in good agreement with the certified values. Comparison with accelerated solvent extraction confirmed high accuracy, and application for a bioconcentration experiment with phenyltins demonstrated the robustness and suitability of the method for routine analyses. The procedure allows fast, reliable, and simple determination of organotin compounds in low-size biological samples, which was demonstrated for bioconcentration experiments. PMID- 11055740 TI - Cigarette smoking among adults--United States, 1998. AB - One of the national health objectives for 2010 is to reduce the prevalence of cigarette smoking among adults to no more than 12% (objective 21.1a). To assess progress toward meeting this objective, CDC analyzed self-reported data from the 1998 National Health Interview Survey (NHIS) Sample Adult Core Questionnaire about cigarette smoking among U.S. adults. This report summarizes the findings of this analysis, which indicate that, in 1998, 24.1% of adults were current smokers. PMID- 11055741 TI - Consequences of delayed diagnosis of Rocky Mountain spotted fever in children- West Virginia, Michigan, Tennessee, and Oklahoma, May-July 2000. AB - Patients with Rocky Mountain spotted fever (RMSF), a tickborne infection caused by Rickettsia rickettsii, respond quickly to tetracycline-class antibiotics (e.g., doxycycline) when therapy is started within the first few days of illness; however, untreated RMSF may result in severe illness and death. Persons aged <10 years have the highest age-specific incidence of RMSF. This report summarizes the clinical course and outcome of RMSF in four children from four regions of the United States and underscores the need for clinicians throughout the United States to consider RMSF in children with rash and fever, particularly those with a history of tick bite or who present during April-September when approximately 90% of RMSF cases occur. PMID- 11055742 TI - Updated recommendations from the Advisory Committee on Immunization Practices in response to delays in supply of influenza vaccine for the 2000-01 season. AB - On July 14, CDC reported a substantial delay in the availability of a proportion of influenza vaccine for the 2000-01 season and the possibility of a vaccine shortage. Since then, resolution of manufacturing problems and improved yields of the influenza A (H3N2) vaccine component have averted a shortage. Although safe and effective influenza vaccine will be available in similar quantities as last year, much of the vaccine will be distributed later in the season than usual. This update provides information on the influenza vaccine supply situation and updated influenza vaccination recommendations by the Advisory Committee on Immunization Practices (ACIP) for the 2000-01 influenza season. PMID- 11055743 TI - Involvement of metabotropic glutamate receptors in ischemia-induced taurine release in the developing and adult hippocampus. AB - Metabotropic glutamate receptors have recently been envisaged as involved in both potentiation and prevention of ischemic and excitotoxic neuronal damage. The release of the inhibitory amino acid taurine is markedly enhanced in ischemia in both the immature and mature mouse hippocampus. The modulation of [3H]taurine release by metabotropic receptor agonists and antagonists was studied in hippocampal slices from developing (7-day-old) and adult (3-month-old) mice using a superfusion system. Agonists of group I, II and III metabotropic glutamate receptors generally reduced the ischemia-induced release in adult animals. In the immature hippocampus the group I agonists (S)-3,5-dihydroxyphenylglycine and (1+/ )-1-aminocyclopentane-trans-1,3-dicarboxylate, which mainly enhance neuronal excitation, potentiated initial taurine release in ischemia. Ionotropic glutamate receptor agonists also enhance the ischemia-induced taurine release in developing mice. This glutamate-activated taurine release may thus constitute an important protective mechanism against excitotoxicity in the immature hippocampus. PMID- 11055744 TI - Phosphatidylcholine metabolism in nuclei of phorbol ester-activated LA-N-1 neuroblastoma cells. AB - The agonist stimulation of a variety of cells results in the induction of specific lipid metabolism in nuclear membranes, supporting the hypothesis of an important role of the lipids in nuclear signal transduction. While the existence of a phosphatidylinositol cycle has been reported in cellular nuclei, little attention has been given to the metabolism of phosphatidylcholine in nuclear signaling. In the present study the metabolism of phosphatidylcholine in the nuclei of neuroblastoma cells LA-N-1 was investigated. The incubation of LA-N-1 nuclei with radioactive choline, phosphocholine or CDP-choline led to the production of labelled phosphatidylcholine. The incorporation of choline and phosphocholine but not CDP-choline was enhanced in nuclei of TPA treated cells. Moreover the presence of choline kinase, phosphocholine cytidylyltransferase and phosphocholine transferase activities were detected in the nuclei and the TPA treatment of the cells stimulated the activity of the phosphocholine cytidylyltransferase. When cells prelabelled with [3H]palmitic acid were stimulated with TPA in the presence of ethanol, an increase of labelled diacylglycerol and phosphatidylethanol in the nuclei was observed. Similarly, an increase of labelled diacylglycerol and phosphatidic acid but not of phosphatidylethanol occurred in [3H]palmitic acid prelabelled nuclei stimulated with TPA in the presence of ethanol. However the production of phosphatidylethanol was observed when the nuclei were treated with TPA in the presence of ATP and GTPgammaS. The stimulation of [3H]choline prelabelled nuclei with TPA also generated the release of free choline and phosphocholine. The results indicate the presence of PLD and probably PLC activities in LA-N-1 nuclei and the involvement of phosphatidylcholine in the production of nuclear lipid second messengers upon TPA stimulation of LA-N-1 cells. The correlation of the disappearance of phosphatidylcholine, the production of diacylglycerol and phosphatidic acid with the stimulation of phosphatidylcholine synthesis in nuclei of TPA treated LA-N-1 suggests the existence of a phosphatidylcholine cycle in these nuclei. PMID- 11055745 TI - Long-lasting ibogaine protection against NMDA-induced convulsions in mice. AB - Ibogaine, a putative antiaddictive drug, is remarkable in its apparent ability to downgrade withdrawal symptoms and drug craving for extended periods of time after a single dose. Ibogaine acts as a non-competitive NMDA receptor antagonist, while NMDA has been implicated in long lasting changes in neuronal function and in the physiological basis of drug addiction. The purpose of this study was to verify if persistent changes in NMDA receptors could be shown in vivo and in vitro after a single administration of ibogaine. The time course of ibogaine effects were examined on NMDA-induced seizures and [3H] MK-801 binding to cortical membranes in mice 30 min, 24, 48, and 72 h post treatment. Ibogaine (80 mg/kg, ip) was effective in inhibiting convulsions induced by NMDA at 24 and 72 hours post administration. Likewise, [3H] MK-801 binding was significantly decreased at 24 and 72 h post ibogaine. No significant differences from controls were found at 30 min or 48 h post ibogaine. This long lasting and complex pattern of modulation of NMDA receptors prompted by a single dose of ibogaine may be associated to its antiaddictive properties. PMID- 11055746 TI - A dual-probe microdialysis study in simultaneously monitoring extracellular pyruvate, lactate, and biogenic amines in gerbil striata during unilateral cerebral ischemia. AB - A dual-probe microdialysis technique coupled with liquid chromatographic assays was developed for the simultaneous monitoring of neurochemicals in gerbil striata during cerebral ischemia. Isocratic separation of lactate and pyruvate was achieved within 5 min whereas the separation of biogenic amines was completed within 30 min. An unilateral ligation was produced by occlusion of the right common carotid artery for 30 mins in anesthetized gerbils to perform a typical focal cerebral ischemia. Microdialysis probes were inserted in both sides of the striata to simultaneously monitor biogenic amines, lactate and pyruvate during cerebral ischemia. Dynamic and comparative changes of these analytes in ipsilateral and contralateral sides of the brain can be simultaneously measured by the assay. The present assay can be used as a research tool to explore neurochemical substances and their relationships during cerebral ischemia. PMID- 11055747 TI - Expression and electrophysiological function of actin in chick cerebellar neurons. AB - Among several monoclonal antibodies obtained by immunizing Balb/c mice with cerebellar synaptic membrane fractions from E20 chick embryos, the antibody, named M35, suppressed Ca-spikes in immature cultured chick cerebellar neurons. M35 immunoprecipitated 43 kDa protein from a 125I-labeled embryonic crude cerebellar membrane fraction. Immunohistochemically, the M35 antigen was expressed most intensively in Purkinje cells, but its expression was limited to highly motile structures at developmental neuronal remodeling. Electrophysiologically, M35 facilitated current responses to AMPA and inhibited the responses to GABA in cultured cerebellar Purkinje neurons. The several peptides derived from the affinity-purified 43 kDa protein were found to have homologous amino acid sequences to non-muscle actins. These results suggest that the antigen recognized by M35 may play an essential role probably as membrane ion channels modulating synaptic functions in not only the development and growth but also the neuronal activity of chick cerebellar Purkinje cells. PMID- 11055748 TI - In vivo evaluation of hippocampal anti-oxidant ability of zonisamide in rats. AB - We evaluated the anti-oxidant property of zonisamide (ZNS) in the rat brain under freely moving conditions by means of in vivo microdialysis of two exogenous nitroxide radicals, 3-carbamoyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (carbamoyl PROXYL) and 3-methoxy carbonyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (PCAM). Time dependent changes in the signal intensities of these exogenous nitroxide radicals obtained from the hippocampal perfusates were observed using an X-band ESR spectrometer at 20-min intervals. The ESR signal intensities of nitroxide radicals decreased exponentially in all animals, which indicates that their half life could be used as a parameter to estimate the decay rate of nitroxide radicals. Nitroxide radicals lose their paramagnetism when exposed to reductants in a biological system. Thus, half-life reflects the in vivo reducing ability. Although the half-life of carbamoyl-PROXYL, which could not pass the blood-brain barrier (BBB), was not changed when compared with the controls, pre-treatment with ZNS significantly shortened the half-life of PCAM, which could pass through the BBB. These findings suggest that the ZNS-induced increase in reducing ability did not occur within the extracellular space, but rather mainly at the neural cell membrane. This study is the first in vivo evaluation of the reducing ability of ZNS in freely moving animals. PMID- 11055749 TI - Temporal expression of mRNAs for neuropoietic cytokines, interleukin-11 (IL-11), oncostatin M (OSM), cardiotrophin-1 (CT-1) and their receptors (IL-11Ralpha and OSMRbeta) in peripheral nerve injury. AB - The mRNA expression pattern of the neuropoietic cytokines, interleukin-11 (IL 11), oncostatin M (OSM) and cardiotrophin-1 (CT-1), and their receptor components (IL-11Ralpha and OSMRbeta) was examined in peripheral nerves on two different types of injury, crush and transection. The IL-11 mRNA increased after nerve damage and immediately returned to control levels. The OSM mRNA expression increased rapidly after nerve injury and relatively high expressions were maintained for at least 14 days. The CT-1 mRNA was not expressed in any time before and after the injury. Interestingly, IL-11Ralpha was expressed in the intact nerve and decreased after injury. The expression of OSMRbeta increased slightly after the injury. Moreover, temporal mRNA expression pattern of these neuropoietic cytokines and receptors was similar between the crushed and transected models. Each neuropoietic cytokine of IL-11, OSM and CT-1 has its own specific temporal mRNA expression pattern, which is also different from those of ciliary neuro-trophic factor (CNTF), leukemia inhibitory factor (LIF) and interleukin-6 (IL-6). These results suggest that all neuropoietic cytokines have distinctive functions in nerve degeneration and repair process in response to peripheral nerve injury. PMID- 11055750 TI - Interference of S-nitrosoglutathione with the binding of ligands to ionotropic glutamate receptors in pig cerebral cortical synaptic membranes. AB - The interactions of S-nitrosoglutathione (GSNO) with the ionotropic glutamate receptors were studied on synaptic membranes isolated from the pig cerebral cortex. GSNO displaced the binding of [3H]glutamate, 3-[(R)-2-carboxypiperazin-4 yl] [3H]propyl-1-phosphonate ([3H]CPP), a competitive N-methyl-D-aspartate (NMDA) antagonist, and [3H]kainate, with IC50 values in the low micromolar range. It failed to displace (S)-5-fluoro-[3H]willardiine, a selective agonist of 2-amino-3 hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors. Reduced and oxidized glutathione were almost as effective as GSNO in glutamate and CPP binding. Of the three, GSNO was the most potent in kainate binding. They all stimulated [3H]dizocilpine binding in a concentration-dependent manner. This effect was additive to that of glycine and not mimicked by NO donors such as S-nitroso-N acetylpenicillamine, 5-amino-3-morpholinyl-1,2,3-oxadiazolium chloride (SIN-1) and nitroglycerin. We assume that GSNO may act as an endogenous ligand at the NMDA and non-NMDA classes of glutamate receptors. In this manner it may facilitate NO transfer and target its delivery to specific sites in these receptors. PMID- 11055751 TI - Identification of macrophage migration inhibitory factor isoforms in bovine brain. AB - In the course of the study of the primary structures and molecular mechanisms of action of immunologically active compounds of the nervous system we have isolated from the soluble fraction of total bovine brain two heat-stable proteins. The purification procedure was mainly based on DEAE-Servacel ion-exchange chromatography and reversed-phase HPLC. The proteins were identified by the N terminal Edman microsequence analysis and database searching as macrophage migration inhibitory factor (MIF). The N-terminal sequences for MIF1 and MIF2 were found to be identical. According to mass spectral analysis, the molecular masses for MIF1 and MIF2 were determined respectively as 12,369.21 and 12,299.7 Da. In addition, we have also isolated a third peptide having the same N-terminal sequence and Mr 9,496.2 that seems to be a proteolytic fragment of MIF. Using p hydroxyphenylpyruvate as a substrate, we have not revealed tautomerase activity of either MIF1 or MIF2. As both the immunologic and enzymatic activities were reported to be expressed by the oligomeric structure of MIF, we suggest that the present study may give additional information on MIF in terms of structural properties of this protein. A comparatively simple purification procedure is presented that may be widely used for simultaneous isolation in one run of MIF isoforms. PMID- 11055752 TI - Regional changes of cyclic 3',5'-guanosine monophosphate in the spinal cord of the rabbit following brief repeated ischemic insults. AB - The regional distribution of cyclic 3',5'-guanosine monophosphate was studied in the lumbosacral segments of the spinal cord of the rabbit under physiological conditions and following brief repeated sublethal ischemic insults. While the basal cGMP level in the gray matter was about 0.120 nmol cGMP/mg wet. wt., the level of cGMP in non-compartmentalized white matter was about half of this value. The highest level of cGMP in the compartmentalized gray matter was found in the dorsal horns, about 0.180 nmol cGMP/mg wet. wt., whereas the level of cGMP was greatly reduced in the ventral horns, reaching one half of the previous value. Multiple sublethal ischemic insults, repeated at 1-h intervals, caused a statistically significant decrease of cGMP in all gray matter regions. While the post-ischemic and post-reperfusion level of cGMP in the dorsal horns remained relatively high in comparison with the intermediate zone and ventral horns, the changes of cGMP level detected in the white matter columns differed considerably and resulted in a statistically significant cGMP increase in the dorsal and ventral columns and, vice versa, a statistically significant decrease of cGMP was found in the lateral columns. PMID- 11055753 TI - Effect of midthoracic spinal cord constriction on catalytic nitric oxide synthase activity in the white matter columns of rabbit. AB - The distribution and changes of catalytic nitric oxid synthase (cNOS) activity in the dorsal, lateral and ventral white matter columns at midthoracic level of the rabbit's spinal cord were studied in a model of surgically-induced spinal cord constriction performed at Th7 segment level and compared with the occurrence of nicotinamide adenine dinucleotide phosphate diaphorase expressing and neuronal nitric oxide synthase immunoreactive axons in the white matter of the control thoracic segments. Segmental and white-column dependent differences of cNOS activity were found in the dorsal (141.5 +/- 4.2 dpm/microm protein), lateral (87.3 +/- 11.5 dpm/microm protein) and ventral (117.1 +/- 7.6 dpm/microm protein) white matter columns in the Th5-Th6 segments and in the dorsal (103.3 +/- 15.5 dpm/microm protein), lateral (54.9 +/- 4.9 dpm/microm protein), and ventral (86.1 +/- 6.8 dpm/microm protein) white matter columns in the Th8-Th9 segments. A surgically-induced constriction of Th7 segment caused a disproportionate response of cNOS activity in the rostrally (Th5-Th6) and caudally (Th8-Th9) located segments in both lateral and ventral white matter columns. While a statistically significant decrease of cNOS activity was detected above the constriction site in the ventral columns, a considerable, statistically significant increase of cNOS activity was noted in the white lateral columns below the site of constriction. It is reasoned that the changes of cNOS activity may have adverse effects on nitric oxide (NO) production in the white matter close to the site of constriction injury, thus broadening the scope of the secondary mechanisms that play a role in neuronal trauma. PMID- 11055754 TI - Ginsenoside Rc and Rg1 differentially modulate NMDA receptor subunit mRNA levels after intracerebroventricular infusion in rats. AB - We investigated the influence of centrally administered ginsenoside on the regulation of mRNA levels of the family of NMDA receptor subtypes (NR1, NR2A, NR2B, NR2C) by in situ hybridization histochemistry in the rat brain. The ginsenosides Rc and Rg1, the major components of ginseng saponin, differentially modulate NMDA receptor subunit mRNA levels in rat brain following prolonged i.c.v.-infusion. Ginsenosides Rc or Rg1 (10 microg/10 microl/hr for 7 days) was infused through preimplanted cannulae connected to osmotic mini-pumps. The level of NR1 mRNA is significantly increased in temporal cortex, caudate putamen, hippocampus, and granule layer of cerebellum in Rg1-infused rats as compared to control group. The level of NR2A mRNA is elevated in the frontal cortex. In contrast, it was decreased in CAI area of hippocampus in Rg1-infused rats. However, there was no significant change of NR1 and NR2A mRNA levels in Rc infused rats. The level of NR2B mRNA is elevated in cortex, caudate putamen, and thalamus in both Rc- and Rg-infused rats. In contrast, NR2B level is decreased in CA3 in Rg1-infused rats. The level of NR2C mRNA is increased in the granule layer of cerebellum in only Rg1 but not Rc infused rats. These results show that structure difference of ginsenoside may diversely affect the modulation of expression of NMDA receptor subunit mRNA after infusion into cerebroventricle in rats. PMID- 11055755 TI - Tuberculin reactivity and the risk of tuberculosis: a review. AB - SETTING: Although various studies have examined the association between tuberculin reactivity and the risk of tuberculosis (TB), this evidence has not been collated and examined to determine the strength and consistency of the association across multiple studies. OBJECTIVE: To review the evidence supporting the association between tuberculin reactivity and the risk of TB. DESIGN: Prospective studies which included raw data on the incidence of TB according to three or more tuberculin reactor categories were located using electronic search methods. The findings of these studies were recalibrated if necessary and compared. RESULTS: All 11 studies identified demonstrated that increased tuberculin skin test (TST) reactivity was associated with an increased risk of TB, and several found that low tuberculin reactivity was associated with a protective effect. The magnitude of the association between TST reactivity and the risk of TB varied substantially. The association between tuberculin reactivity and the risk of TB was greater among studies that reported a lower incidence of TB among the smallest tuberculin reactor category. CONCLUSION: All studies reviewed support a positive association between tuberculin reactivity and the risk of TB. However, this review found a substantial degree of variation in the extent of increased risk associated with larger tuberculin reactions. PMID- 11055756 TI - The impact of directly-observed treatment on the epidemiology of tuberculosis in Beijing. AB - SETTING: Fully supervised chemotherapy, or directly observed treatment (DOT), for newly detected smear-positive cases in Beijing, has been successfully implemented for two decades. OBJECTIVE: To evaluate the progress made in tuberculosis control, and in particular to evaluate the impact of DOT on tuberculosis epidemiology in Beijing. DESIGN: Epidemiological parameters on tuberculosis, consisting of mortality, prevalence, notification rate, tuberculous meningitis in children and initial drug resistance rate, were collected and analysed. Their trends were evaluated and compared with DOT implemented for new smear-positive cases in Beijing from 1978 to 1996. RESULTS: The coverage of DOT for new smear positive pulmonary tuberculosis cases has increased from 10% in 1978 to more than 90% since 1990. Since DOT was introduced in 1978, mortality from tuberculosis has declined by an average of more than 7% per year. The reduction rate of 17.2%, and the rates of chronic cases and tuberculous meningitis in children decreased dramatically. The rate of newly registered smear-positive cases decreased from 18.9/100000 in 1986 to 7.3/100000 in 1996, giving an average annual reduction rate of 9.1 during this period. Initial resistance to isoniazid and streptomycin decreased from respectively 13.9% and 12.3% in 1978-1979 to 4.2% and 5.8% by 1996. The level of multidrug resistance was low and stable, at 0.8% in 1996. CONCLUSION: The experience of the Beijing tuberculosis control programme convincingly demonstrates that it is possible to improve the epidemiological situation rapidly in a low-income country, at very low cost and in a manner that is self-sufficient and sustainable. PMID- 11055757 TI - Twenty-year trend of chronic excretors of tubercle bacilli based on the nationwide tuberculosis prevalence surveys in Korea, 1975-1995. AB - SETTING: A study of chronic excretors of tubercle bacilli (chronic cases) based on the nationwide random sample surveys of tuberculosis prevalence conducted in Korea from 1975 through 1995. OBJECTIVE: To investigate the temporal trend of the prevalence of chronic cases, and to match these with treatment outcomes and drug resistance rates. DESIGN: Bacillary cases were classified by history of chemotherapy into new (those who denied a history of chemotherapy), non-chronic (those who had taken chemotherapy for less than 2 years) and chronic cases (those who had taken chemotherapy for more than 2 years). RESULTS: Chronic cases decreased from 107 to 12 per 100000 population (annual rate of reduction [ARR] 11.89%) over the 20-year period. The ARR of chronic cases was significantly greater than that of new cases, and accelerated from 1985 (ARR 15.83%), after the application of short course chemotherapy. Rates of overall drug resistance rates increased up to 1980, and those of multidrug resistance up to 1985, followed by a decrease thereafter. A reduction in chronic cases was observed even during the period of increase in drug resistance (including multidrug resistance). CONCLUSION: The prevalence of chronic tuberculosis cases has decreased due to improvements in overall treatment outcome. PMID- 11055758 TI - Is TB contact screening relevant in a developing country setting? Experiences from eastern Nepal, 1996-1998. AB - SETTING: A tuberculosis programme run by a non-governmental organisation in eight hill and mountain districts of eastern Nepal. OBJECTIVE: To assess the impact of contact screening on case-finding. DESIGN: A retrospective cohort study of contacts of smear-positive, smear-negative and extra-pulmonary tuberculosis patients diagnosed and registered during 1996-1998 ('index cases'). Contacts, defined as household members identified by index cases, were screened by sputum examination; two positive smears were taken to indicate smear-positive pulmonary disease. RESULTS: Approximately 50% (668) of registered cases identified contacts; 75% (2298) of the contacts identified provided one or more sputum specimens. An overall smear-positive case yield of 0.61% (14) was obtained from contacts tested, all except one of which were contacts of smear-positive index cases. For smear-positive index cases with a smear grading of > or = 2+, the yield was 7.2 times greater (P = 0.04) than for those with a grading of 1+. CONCLUSION: In this setting, sputum examination of household contacts of smear negative and extrapulmonary tuberculosis cases is not justified. Further assessment is needed to evaluate the utility of testing contacts of smear positive cases without symptom screening, and whether cost effectiveness can be improved by restricting testing to contacts of cases with high bacterial (> or = 2+) loads. PMID- 11055759 TI - Tuberculosis infection in an Aboriginal (First Nations) population of Canada. AB - BACKGROUND: The incidence of active tuberculosis (TB) among the Cree, an Aboriginal population of Canada, is dropping, but it remains three times that of the general population. We analyzed data from tuberculin skin test (TST) surveys to determine estimates of prevalence of infection and annual risk of infection (ARI) in this population. METHODS: TST surveys targeting 12-year-old students were conducted annually from 1993 to 1998. Students with no record of previous positive TST (> or = 10 mm) were offered TST (5 TU PPD-T). Data collected included result of previous TST reading for all students, readings of TSTs performed (mm induration) and BCG (bacille Calmette-Guerin) vaccination status for those positive on TST. RESULTS: A total of 1274 children were screened (participation rate 94%). TST reaction size frequency distribution plots a bimodal curve. The prevalence of infection among 12 year olds was 15.3% over this period. ARI estimates range from 0.6 to 2.4% (average ARI 1.4%). A significant downward linear trend in ARI was observed over the period (P < 0.001). DISCUSSION: Calculated ARI may be over-estimated due to prior BCG vaccination; however, the trend in ARI confirms decreasing transmission of TB infection. Better knowledge of human immunodeficiency virus seroprevalence among pregnant women is needed to complete the evaluation of the BCG program. PMID- 11055760 TI - Causes and costs of hospitalization of tuberculosis patients in the United States. AB - OBJECTIVE: To examine the costs, lengths of stay and patient characteristics associated with tuberculosis (TB) hospitalizations. METHODS: A prospective cohort study of 1493 TB patients followed from diagnosis to completion of therapy at 10 public health programs and area hospitals in the US. The main outcome measures were the following: 1) occurrence, 2) cost, and 3) length of stay of TB-related hospitalizations. RESULTS: There were 821 TB-related hospitalizations among the study participants; 678 (83%) were initial hospitalizations and 143 (17%) were hospitalizations during the treatment of TB. Patients infected with human immunodeficiency virus (HIV) (OR 1.8, 95% CI 1.2-2.6), and homeless patients (OR, 1.7 95% CI 1.1-2.8) were at increased risk of being hospitalized at diagnosis. Homeless patients (RR 2.5, 95%CI 1.5-4.3), patients who used alcohol excessively (RR 1.9, 95% CI 1.2-3.0), and patients with multidrug-resistant TB (RR 5.7, 95% CI 2.7-11.8) were at increased risk of hospitalization during treatment. The median length of stay varied from 9 to 17 days, and median costs per hospitalization varied from $6441 to $12968 among the sites. CONCLUSION: Important social factors, HIV infection, and local hospitalization practice patterns contribute significantly to the high cost of TB-related hospitalizations. Efforts to address these specific factors are needed to reduce the cost of preventable hospitalizations. PMID- 11055761 TI - Surveillance of anti-tuberculosis drug resistance: results of the 1998/1999 proficiency testing in Italy. SMIRA (Italian Multicentre Study on Antituberculosis Drug Resistance) Study Group. AB - OBJECTIVE: To determine the accuracy of drug-susceptibility testing (DST) for isoniazid, rifampicin, ethambutol and streptomycin in a provisional network of 22 regional laboratories in Italy. METHODS: Methods, definitions and reference Mycobacterium tuberculosis strains were derived from the WHO/IUATLD Global Project on Anti-tuberculosis Drug Resistance Surveillance. The laboratories were selected based on technical skills required by the project, the number of DST performed annually and geographic localisation. The results (sensitive/resistant strain) were compared with the gold standard (global project results). Sensitivity (ability to detect true resistance), specificity (ability to detect true susceptibility), positive predictive values for resistance and susceptibility, efficiency and reproducibility were calculated in two rounds. RESULTS: Eighteen of 22 laboratories completed the first round of proficiency testing for the four drugs. Sensitivity was 76.6%, specificity 97.2%, predictive value of a resistant test 89.8% and of a susceptible test 86.8%, efficiency 87.8% and reproducibility 92.8%. A second round was performed by all those laboratories that did not achieve > or = 90% agreement with the results of the Global Project. Overall, after the second round, all the parameters except specificity improved, exceeding 90%. CONCLUSIONS: A network of 15 regional laboratories that fulfil the quality criteria for determining the susceptibility of M. tuberculosis to the four primary antituberculosis drugs was established in Italy. PMID- 11055762 TI - Surveillance of drug-resistant tuberculosis and molecular evaluation of transmission of resistant strains in refugee and non-refugee populations in North Eastern Kenya. AB - SETTING: Three refugee camp complex clinics and an adjacent non-refugee treatment centre in North-Eastern Kenya. OBJECTIVES: To use conventional and molecular epidemiology tools to determine: 1) the prevalence of drug resistance in newly diagnosed patients with smear-positive pulmonary tuberculosis in refugee and non refugee populations; 2) risk factors for resistance in the two populations; and 3) whether IS6110 restriction fragment length polymorphism (RFLP) and spoligotyping showed similarities in DNA fingerprinting patterns of drug resistant isolates that could infer transmission within and between the two populations. RESULTS: Of 241 isolates from the camps, 44 (18.3%) were resistant to one or more drugs, seven of which (2.9%) were multidrug-resistant TB (MDR-TB). Of 88 isolates from the non-refugees, five (5.7%) were resistant to one or more drugs without MDR-TB. Drug resistance was higher in the camps than in the non refugee population (OR = 3.7; 95%CI 1.42-9.68; P < 0.007). Resistance was significantly higher in one camp compared with the other two, despite a comparable ethnic distribution. Unusually, females were more associated with drug resistance than their male counterparts in both populations (OR = 2.3; 95%CI 1.2 4.8; P = 0.008). There was evidence of transmission of streptomycin-resistant strains in the refugee population. DNA fingerprints of resistant strains from the non-refugee population were unique and different from those in the refugee camps. CONCLUSION: The observed high levels of drug resistance and MDR-TB, combined with evidence of transmission of strains resistant to streptomycin in the refugee population, suggest a need for strengthened TB control programmes in settings with a high risk of developing drug-resistant strains. PMID- 11055763 TI - Completeness and timeliness of treatment initiation after laboratory diagnosis of tuberculosis in Gaborone, Botswana. AB - SETTING: Gaborone, the capital of Botswana. OBJECTIVE: To determine the time from positive sputum smear microscopy for acid-fast bacilli (AFB) to initiation of therapy, and to identify risk factors for delays. DESIGN: Retrospective cohort study of medical records and surveillance data for patients with positive smear microscopy and newly diagnosed tuberculosis (TB) from January to May 1997. Treatment delay was defined as more than 2 weeks from the first positive sputum smear to the initiation of TB treatment. RESULTS: Of 127 patients identified, 15 (11.8%) had treatment delay, 13 (10.2%) had an incomplete workup (only one smear performed) and were not registered for TB treatment, and six (4.5%) had two or more positive smears but were not registered for TB treatment. Risk factors for treatment delay or non-registration included TB patients who had been diagnosed in a hospital outpatient setting vs. a clinic (RR 2.9, 95% CI 1.2-3.6, P = 0.02), or in a high volume vs. low volume clinic (RR 2.2, 95% CI 1.2-5.3, P = 0.01). CONCLUSION: More than a quarter of the smear-positive TB patients identified had treatment delay or no evidence of treatment initiation. Proper monitoring of laboratory sputum results and suspect TB patient registers could potentially reduce treatment delays and patient loss. PMID- 11055764 TI - Transmission of Mycobacterium tuberculosis among employees in a US government office, Gaborone, Botswana. AB - SETTING: A US government office located in Botswana where two office employees, one negative and one positive for the human immunodeficiency virus (HIV), were diagnosed with pulmonary tuberculosis (TB) in January 1998. One employee had been symptomatic with untreated laryngeal TB for 8 months. OBJECTIVE: To determine the extent of and risk factors for TB transmission in the office. METHODS: Office contacts were interviewed and a tuberculin skin test (TST) was performed. A positive TST was defined as > or = 10 mm induration for employees from countries where TB is highly endemic, and as > or = 5 mm induration for those from low prevalence counties. RESULTS: Of 79 office contacts investigated, 54/57 (94.7%) born in high TB prevalence countries had a positive TST compared with 4/22 (18.2%) from low prevalence countries (RR 5.1, 95% CI 2.1-12.7, P < 0.001). Of 20 US-born contacts, three (15%) had documented TST conversion, two of whom were co workers of the employee with laryngeal TB. Isolates of Mycobacterium tuberculosis from the TB cases had matching DNA fingerprints. CONCLUSION: Delayed diagnosis in a setting of high TB prevalence may have contributed to transmission within a US government office located in Botswana. Transmission may have been underestimated due to the high background prevalence of tuberculous infection in the population. Recent tuberculous transmission to persons living with HIV infection may be playing an important role in the escalating TB epidemic in Africa. PMID- 11055765 TI - Mortality rates and recurrent rates of tuberculosis in patients with smear negative pulmonary tuberculosis and tuberculous pleural effusion who have completed treatment. AB - SETTING: Queen Elizabeth Central Hospital, Blantyre, and Zomba Central Hospital, Zomba, Malawi. OBJECTIVE: To follow-up human immunodeficiency virus (HIV) seropositive and HIV-seronegative patients with smear-negative pulmonary tuberculosis (PTB) and pleural TB who had completed treatment with two different regimens in Blantyre and Zomba, and to assess rates of mortality and recurrent TB. DESIGN: Patients with smear-negative and pleural TB who had completed 8 months ambulatory treatment in Blantyre or 12 months standard treatment in Zomba and who were smear and culture negative for acid-fast bacilli at the completion of treatment were actively followed every 4 months for a total of 20 months. RESULTS: Of 248 patients, 150 with smear-negative PTB and 98 with pleural TB, who completed treatment and were enrolled, 205 (83%) were HIV-positive. At 20 months, 145 (58%) patients were alive, 85 (34%) had died and 18 (7%) had transferred out of the district. The mortality rate was 25.7 per 100 person-years, with increased rates strongly associated with HIV infection and age >45 years. Forty-nine patients developed recurrent TB. The recurrence rate of TB was 16.1 per 100 person-years, with increased rates strongly associated with HIV infection, having smear-negative PTB and having received 'standard treatment'. CONCLUSION: High rates of mortality and recurrent TB were found in patients with smear-negative PTB and pleural effusion during 20 months of follow-up. TB programmes in sub Saharan Africa must consider appropriate interventions, such as co-trimoxazole and secondary isoniazid prophylaxis, to reduce these adverse outcomes. PMID- 11055766 TI - Diagnosis of tuberculous pleuritis by the measurement of soluble interleukin 2 receptor in pleural fluid. AB - OBJECTIVE: As soluble interleukin-2 receptor (sIL-2R) is a marker of T-lymphocyte activation, we sought to determine whether its measurement in pleural fluid is diagnostically useful in tuberculous pleurisy. DESIGN: We compared the concentrations of sIL-2R in pleural samples of 23 patients with tuberculous pleurisy and 109 patients with non-tuberculous effusions (34 malignant, 34 parapneumonic, 27 transudates and 14 miscellaneous). sIL-2R was measured by a commercial ELISA test and its performance was evaluated using receiver operating characteristic (ROC) analysis. RESULTS: The mean values of pleural sIL-2R were 9179 U/mL in patients with tuberculous pleurisy vs 3664 U/mL in patients with malignancy, 2603 U/mL in patients with parapneumonic effusions, 1016 U/mL in patients with transudates, and 1906 U/mL in patients with miscellaneous diseases (P < 0.0001). A ROC curve identified the best cut-off at 4700 U/mL, yielding measures for sensitivity (0.91), specificity (0.94) and accuracy (0.94). CONCLUSIONS: The results of this pilot study suggest that pleural sIL-2R appears to be clinically useful for differentiating between tuberculous and non tuberculous pleural effusions. PMID- 11055767 TI - Efficacy of culture filtrate protein preparations from Indian isolates of M. tuberculosis to activate T cells derived from healthy donors. AB - SETTING: While culture filtrate proteins (CFPs) of Mycobacterium tuberculosis appear to be good vaccine candidates for tuberculosis, only CFPs derived from certain popular laboratory strains of M. tuberculosis have been studied for this purpose. OBJECTIVE: To compare the relative efficacies of CFP preparations from two laboratory strains and four contemporary clinical isolates of M. tuberculosis to induce T-cell activation. DESIGN: CFPs were isolated from six strains of M. tuberculosis and were used to induce 1) T-cell proliferation, 2) IFN-gamma secretion, and 3) IL-12 secretion from peripheral blood derived mononuclear cell (PBMC) preparations from 33 healthy donors. RESULTS: Significant amounts of IL-12 were spontaneously secreted by PBMC preparations; CFP preparations from two clinical isolates (JNU-7 and JNU-51) significantly boosted this response. All six CFP preparations induced IFN-gamma secretion by PBMCs, but those from two contemporary strains of M. tuberculosis (JNU-7 and JNU-22) were most effective in this regard. The effect of CFPs from JNU-7 and JNU-22 was significantly better than those from the laboratory strains (H37Ra and Erdman). Similar results were obtained with the T-cell proliferation parameter. CONCLUSION: These results suggest that CFPs derived from selected clinical isolates of M. tuberculosis may outperform those of standard laboratory strains, and may therefore be a better source of potential candidates for a tuberculosis vaccine. PMID- 11055768 TI - Three stories about green eggs. Story one: Green Eggs and freedom. PMID- 11055769 TI - The power of knowledge to effect change: the 1997 Philippines nationwide tuberculosis prevalence survey. PMID- 11055770 TI - Incorrect estimation of TB drug resistance in Israel, probably due to recruitment and collection biases. PMID- 11055771 TI - Isopathy: nostrum or nocebo? Where now? PMID- 11055772 TI - Is homeopathic 'immunotherapy' effective? A double-blind, placebo-controlled trial with the isopathic remedy Betula 30c for patients with birch pollen allergy. AB - The objective of the study was to examine the effect of the homeopathic remedy Betula 30c vs. placebo for patients with birch pollen allergy. A double-blind, randomized, placebo-controlled trial was carried out. Tablets were given for 4 weeks during the birch pollen season. The setting was Oslo, Norway, May 1995. Patients were aged between 18 and 50 y; 32 patients received Betula 30c tablets and 34 patients received placebo tablets. The main outcome measure was the total score of 17 different allergy symptoms. Daily total scores were calculated, as well as differences and ratios between the run-in and the following time periods. Point estimates of the median difference between the experimental and placebo groups, with their 95% confidence intervals, were the main measure of effect. No statistically significant difference between the groups was found during the first and last period of May. However, from 8 to 18 May, a clinically interesting difference was revealed between the groups, those receiving Betula 30c having fewer and less serious symptoms. For some days these differences were statistically significant. Surprisingly, this group reported more aggravation from the tablets than did the placebo group. With a statistical power of 70% for a defined clinically interesting difference (25%), the present results indicate that treatment with Betula 30c during the pollen season deserves further attention. PMID- 11055773 TI - No beneficial effect of isopathic prophylactic treatment for birch pollen allergy during a low-pollen season: a double-blind, placebo-controlled clinical trial of homeopathic Betula 30c. AB - The objective of this research was to determine if the homeopathic medicine Betula 30c is more effective than placebo at reducing symptoms of pollen allergy in patients sensitive to birch pollen. It was a double-blind, randomized, placebo controlled trial. Tablets were given both as a prophylactic agent, once a week four weeks before the pollen season and as an acute remedy during the pollen season. The study was done in Oslo, Norway, in May 1996 and involved 73 children, adolescents and young adults from 7 to 25 y of age. Allergy-symptoms were assessed on a visual analogue scale (VAS) by patients or parents. Main outcome measure was the median (with its 95% confidence interval) of the symptom scores for all the treated patients, each day during a 10-day period. The pollen count was very low in 1996, only three days were high enough to provoke allergic symptoms. Surprisingly, the verum treated patients fared worse than the placebo group; they used more rescue medication and had higher symptom scores during these three days. Homeopaths might attribute the findings to a putative aggravation response, but the results certainly do not lend support to the usefulness of the tested prophylactic approach, under conditions of low allergen exposure. PMID- 11055774 TI - Evaluation of 2LHERP in preventing recurrences of genital herpes. Institut International 3IDI. AB - The objective of the study was to evaluate the effectiveness of a homeopathic complex in terms of intensity of attacks and duration of remission between attacks of genital herpes. Fifty three patients aged 18 or over with a minimum of four attacks annually were followed in this open multicentre study in a primary care setting. The principal parameters analysed were: frequency of attacks; intensity of symptoms, during treatment and/or after stopping treatment; treatment tolerance. Eighty-two percent of patients treated for recurrent genital herpes benefited. In 41% of cases, there was no recurrence after the first treatment with follow-up of between 8 and 50 months. In 32% of patients, one or two relapses, in 9% of patients recurrences continued but with reduced frequency and intensity. PMID- 11055775 TI - Audit of outcome in 829 consecutive patients treated with homeopathic medicines. AB - An audit was conducted of 829 consecutive patients presenting for homeopathic treatment of a chronic illness, conventional treatment had either failed, plateaued in effect, or was contraindicated by adverse effects, age or condition of the patient. Of the 829 patients, 503 (61%) had a sustained improvement from homeopathic treatment, of these: * 357 patients (43%) had an excellent response; * 146 patients (18%) had a good response; * 6 patients (0.8%) became worse. * 233 patients (28%) were lost to follow-up. PMID- 11055776 TI - Inter-rater reliability of symptom repertorisation: a pragmatic empirical study. AB - OBJECTIVE: To determine the extent to which two homeopaths agree on whether symptoms reported by patients in a proving are possibly associated with Mercurius solubilis. DESIGN: Blinded, inter-rater reliability study. PARTICIPANTS: 104 subjects in a randomised, double-blind mercury proving. OUTCOME MEASURES: 557 symptom episodes spontaneously reported by subjects were classified as 'mercury' or 'not mercury' by two homeopaths working blind to each other's conclusions and to patient allocation. RESULTS: Initial agreement between homeopaths was 70.2%, a kappa of 0.39, (95% CI 0.31, 0.47). Some disagreements appear to have resulted from differing interpretations of the study instructions. After suitable correction, agreement was 76.5% and kappa 0.56 (95% CI 0.49, 0.63). CONCLUSIONS: The study homeopaths had only a moderate level degree of agreement greater than that expected by chance. The main factor seems to have been differences between data from different sources. There is an urgent need for more research on the methods of choosing homoeopathic medicines in order to improve the reliability and validity of homoeopathic diagnoses. PMID- 11055777 TI - More trials, fewer placebos, please. AB - The argument that randomized placebo-controlled trials of homeopathy might usefully be replaced by observational studies, audit and quality-of life assessment is considered. Randomized equivalence and patient-preference trials are proposed as more informative alternatives. They have the merit of providing hard information for health services on the comparative value of treatments, and can facilitate internal comparisons of competing homeopathic methods. The pragmatic approach also allows clinical change during the homeopathic treatment of chronic disease to be assessed without the time constraints usually imposed by placebo controls. PMID- 11055778 TI - The toxicology of Myrmecia nigrocincta, an Australian ant. AB - The biology, behaviour and venom of the ant Myrmecia nigrocincta are described. Symptoms of 15 cases of envenomation are described. PMID- 11055779 TI - The toxicology of Heloderma suspectum: the Gila monster. AB - The biotoxicology of Heloderma suspectum, the Gila monster, is presented in order to complement the homeopathic provings. PMID- 11055780 TI - Liga Medicorum Homoeopathica Internationalis: Budapest, Hungary, 13-17 May, 2000. PMID- 11055781 TI - Nondistorting flattening maps and the 3-D visualization of colon CT images. AB - In this paper, we consider a novel three-dimensional (3-D) visualization technique based on surface flattening for virtual colonoscopy. Such visualization methods could be important in virtual colonoscopy because they have the potential for noninvasively determining the presence of polyps and other pathologies. Further, we demonstrate a method that presents a surface scan of the entire colon as a cine, and affords the viewer the opportunity to examine each point on the surface without distortion. We use certain angle-preserving mappings from differential geometry to derive an explicit method for flattening surfaces obtained from 3-D colon computed tomography (CT) imagery. Indeed, we describe a general method based on a discretization of the Laplace-Beltrami operator for flattening a surface into the plane in a conformal manner. From a triangulated surface representation of the colon, we indicate how the procedure may be implemented using a finite element technique, which takes into account special boundary conditions. We also provide simple formulas that may be used in a real time cine to correct for distortion. PMID- 11055782 TI - Correction of MR kappa-space data corrupted by spike noise. AB - Magnetic resonance images are reconstructed from digitized raw data, which are collected in the spatial-frequency domain (also called kappa-space). Occasionally, single or multiple data points in the k-space data are corrupted by spike noise, causing striation artifacts in images. Thresholding methods for detecting corrupted data points can fail because of small alterations, especially for data points in the low spatial frequency area where the k-space variation is large. Restoration of corrupted data points using interpolations of neighboring pixels can give incorrect results. We propose a Fourier transform method for detecting and restoring corrupted data points using a window filter derived from the striation-artifact structure in an image or an intermediate domain. The method provides an analytical solution for the alteration at each corrupted data point. It can effectively restore corrupted kappa-space data, removing striation artifacts in images, provided that the following three conditions are satisfied. First, a region of known signal distribution (for example, air background) is visible in either the image or the intermediate domain so that it can be selected using a window filter. Second, multiple spikes are separated by the full-width at half-maximum of the point spread function for the window filter. Third, the magnitude of a spike is larger than the minimum detectable value determined by the window filter and the standard deviation of kappa-space random noise. PMID- 11055783 TI - An inverse problem approach to the correction of distortion in EPI images. AB - Magnetic resonance imaging using the echo planar imaging (EPI) technique is particularly sensitive to main (B0) field inhomogeneities. The primary effect is geometrical distortion in the phase encoding direction. In this paper, we present a method based on the conjugate gradient algorithm to correct for this geometrical distortion, by solving the EPI imaging equation. Two versions are presented: one that attempts to solve the full four-dimensional (4-D) imaging equation, and one that independently solves for each profile along the blip encoding direction. Results are presented for both phantom and in vivo brain EPI images and compared with other proposed correction methods. PMID- 11055784 TI - Real-time image reconstruction for spiral MRI using fixed-point calculation. AB - Because spiral magnetic resonance imaging (MRI) is more robust to motion artifacts than echo planar imaging (EPI), spiral imaging method is more suitable in real-time imaging applications where dynamic processes are to be observed. The major hurdle to use spiral imaging method in real-time applications is its slow reconstruction speed. Since spiral trajectories do not sample data on rectilinear grids, raw data must be regridded before inverse fast Fourier transform (FFT). At present, the computational cost for the spiral reconstruction algorithm is still too high and it is not fast enough to achieve the minimum speed requirement of 20 frames/s for real-time imaging applications. In this paper, we propose to replace floating-point calculations with fixed-point calculations in the reconstruction algorithm to remove the computational bottlenecks. To overcome the quantization and round-off errors introduced by fixed-point calculations, we devise a method to find the optimal precision for the fixed-point representation. Adding with a highly efficient vector-radix two-dimensional (2-D) FFT algorithm and modifications to speed up the gridding convolution, we have cut the reconstruction time by 42% and achieved real-time reconstruction at 30 frames/s for 128 x 128 matrices on low-cost PC's. PMID- 11055785 TI - Interpolation of 3-D binary images based on morphological skeletonization. AB - In this paper, the morphological skeleton interpolation (MSI) algorithm is presented. It is an efficient, shape-based interpolation method used for interpolating slices in a three-dimensional (3-D) binary object. It is based on morphological skeletonization, which is used for two-dimensional (2-D) slice representation. The proposed morphological skeleton matching process provides translation, rotation, and scaling information at the same time. The interpolated slices preserve the shape of the original object slices, when the slices have similar shapes. It can also modify the shape of an object when the successive slices do not have similar shapes. Applications on artificial and real data are also presented. PMID- 11055786 TI - Morphology-based three-dimensional interpolation. AB - In many medical applications, the number of available two-dimensional (2-D) images is always insufficient. Therefore, the three-dimensional (3-D) reconstruction must be accomplished by appropriate interpolation methods to fill gaps between available image slices. In this paper, we propose a morphology-based algorithm to interpolate the missing data. The proposed algorithm consists of several steps. First, the object or hole contours are extracted using conventional image-processing techniques. Second, the object or hole matching issue is evaluated. Prior to interpolation, the centroids of the objects are aligned. Next, we employ a dilation operator to transform digital images into distance maps and we correct the distance maps if required. Finally, we utilize an erosion operator to accomplish the interpolation. Furthermore, if multiple objects or holes are interpolated, we blend them together to complete the algorithm. We experimentally evaluate the proposed method against various synthesized cases reported in the literature. Experimental results show that the proposed method is able to handle general object interpolation effectively. PMID- 11055787 TI - An imaging system with calibrated color image acquisition for use in dermatology. AB - We propose a novel imaging system useful in dermatology, more precisely, for the follow-up of patients with an increased risk of skin cancer. The system consists of a Pentium PC equipped with an RGB frame grabber, a three-chip charge coupled devices (CCD) camera controlled by the serial port and equipped with a zoom lens and a halogen annular light source. Calibration of the imaging system provides a way to transform the acquired images, which are defined in an unknown color space, to a standard, well-defined color space called sRGB. sRGB has a known relation to the CIE1 XYZ and CIE L*a*b* colorimetric spaces. These CIE color spaces are based on the human vision, and they allow the computation of a color difference metric called CIE deltaE*ab, which is proportional to the color difference, as seen by a human observer. Several types of polynomial RGB to sRGB transforms will be tried, including some optimized in perceptually uniform color spaces. The use of a standard and well-defined color space also allows meaningful exchange of images, e.g., in teledermatology. The calibration procedure is based on 24 patches with known color properties, and it takes about 5 minutes to perform. It results in a number of settings called a profile that remains valid for tens of hours of operation. Such a profile is checked before acquiring images using just one color patch, and is adjusted on the fly to compensate for short term drift in the response of the imaging system. Precision or reproducibility of subsequent color measurements is very good with (deltaE*ab) = 0.3 and deltaE*ab < 1.2. Accuracy compared with spectrophotometric measurements is fair with (deltaE*ab) = 6.2 and deltaE*ab < 13.3. PMID- 11055788 TI - Normalization of local contrast in mammograms. AB - Equalizing image noise has been shown to be an important step in automatic detection of microcalcifications in digital mammograms. In this study, an accurate adaptive approach for noise equalization is presented and investigated. No additional information obtained from phantom recordings is involved in the method, which makes the approach robust and independent of film type and film development characteristics. Furthermore, it is possible to apply the method on direct digital mammograms as well. In this study, the adaptive approach is optimized by investigating a number of alternative approaches to estimate the image noise. The estimation of high-frequency noise as a function of the grayscale is improved by a new technique for dividing the grayscale in sample intervals and by using a model for additive high-frequency noise. It is shown that the adaptive noise equalization gives substantially better detection results than does a fixed noise equalization. A large database of 245 digitized mammograms with 341 clusters was used for evaluation of the method. PMID- 11055789 TI - Interpolation revisited. AB - Based on the theory of approximation, this paper presents a unified analysis of interpolation and resampling techniques. An important issue is the choice of adequate basis functions. We show that, contrary to the common belief, those that perform best are not interpolating. By opposition to traditional interpolation, we call their use generalized interpolation; they involve a prefiltering step when correctly applied. We explain why the approximation order inherent in any basis function is important to limit interpolation artifacts. The decomposition theorem states that any basis function endowed with approximation order can be expressed as the convolution of a B-spline of the same order with another function that has none. This motivates the use of splines and spline-based functions as a tunable way to keep artifacts in check without any significant cost penalty. We discuss implementation and performance issues, and we provide experimental evidence to support our claims. PMID- 11055790 TI - The effect of image distortion on 3-D reconstruction of coronary bypass grafts from angiographic views. AB - Three-dimensional (3-D) reconstructions of coronary bypass grafts performed from X-ray angiographic images may become increasingly important for the investigation of damaging mechanical stresses imposed to these vessels by the cyclic movement of the heart. Contrary to what we had experienced with coronary arteries, appreciable reconstruction artifacts frequently occur with grafts. In order to verify the hypothesis that those are caused by distortions present in the angiographic images (acquired with image intensifiers), we have implemented a grid correction technique in our 3-D reconstruction method and studied its efficiency with phantom experiments. In this article, the nature of the encountered artifacts and the way in which the dewarping correction eliminates them are illustrated by a phantom experiment and by the reconstruction of a real coronary bypass vein graft. PMID- 11055791 TI - Segmenting skin lesions with partial-differential-equations-based image processing algorithms. AB - In this paper, a partial-differential equations (PDE)-based system for detecting the boundary of skin lesions in digital clinical skin images is presented. The image is first preprocessed via contrast-enhancement and anisotropic diffusion. If the lesion is covered by hairs, a PDE-based continuous morphological filter that removes them is used as an additional preprocessing step. Following these steps, the skin lesion is segmented either by the geodesic active contours model or the geodesic edge tracing approach. These techniques are based on computing, again via PDEs, a geodesic curve in a space defined by the image content. Examples showing the performance of the algorithm are given. PMID- 11055792 TI - Quantitative and kinetic evolution of wound healing through image analysis. AB - To define a healing function based on parameters measured on digitized images of wounds, and to use it to compare the rate of healing of two skin graft donor sites, one treated with petrolatum gauze (Pg) and the other with a topical preparation containing alginates (A). Digital photographs of donor sites (depth 0.6 mm) taken every two days between day 6 and day 12 were analyzed blind using the same algorithm, following changes in color and homogeneity. Analysis of variance was used to identify those parameters that changed during healing. The healing function was constructed using measurements made in six patients (group 1) randomly chosen from ten requiring skin grafts, and was applied and validated using data from the remaining four patients (group 2). The results given by this healing function were compared with those provided by principal component analysis. The most significant healing parameters were those measuring wound homogeneity, and our healing function reflects how these change with time. The time-dependent curves of the function calculated for groups 1 and 2 matched well enough to be considered as being derived from the same set of measurements. The results given by this healing function explained, by analogy, the meaning of the first principal component of principal component analysis. From day 6 to day 12, the healing function followed the same time-course for the Pg and A treatments, but healing was achieved significantly earlier (4 days, p < 0.03) with A. This suggests that the effect of A on wound healing is achieved in the first six days, before the visual changes from epidermalization are analyzable. PMID- 11055793 TI - Cross-contamination of cell lines in culture. PMID- 11055794 TI - Epigenetic mechanisms of the carcinogenic effects of xenobiotics and in vitro methods of their detection. AB - Carcinogenesis is associated with various epigenetic mechanisms, which can alter intra- and intercellular communication and gene expression and thus affect cytokinetics, i.e. regulation of cell proliferation, differentiation, and apoptosis. These processes lead to a loss of homeostatic control. In addition to "classical" epigenetic events such as DNA methylation and histone acetylation, the major mechanisms include changes in concentrations of signal molecules (hormones, growth factors, fatty acids, etc.), modulation of cell receptors and drug-, hormone- and fatty acid-metabolizing enzymes, oxidative stress, and interference with intracellular signal transduction pathways. Multidisciplinary and multibiomarker approach is necessary for setting up a battery of specific biochemical, molecular, and cellular in vitro methods detecting the epigenetic carcinogenic potential of individual chemicals or their environmental mixtures. This approach is based on studies of modes of action of xenobiotics at various levels, including the molecular mechanisms and modulations of cytokinetics, each of them having its specific predictive value. PMID- 11055795 TI - Cytogenetic investigations on microwaves emitted by a 455.7 MHz car phone. AB - The chromosome aberration or sister chromatid exchange frequency was determined in 455.7 MHz microwave-exposed human lymphocytes and in lymphocytes that were subsequently exposed to MMC or X-rays. The exposure was performed by placing the cells at 5 cm from the antenna of a car phone. In this way the specific absorption ratio was approximately 6.5 W/kg. The temperature and humidity was kept constant during the experiments. No statistically significant difference was found between microwave-exposed and unexposed control samples. When the microwave exposure was followed by exposure to MMC, some differences were found between the combined treatments and the MMC treatments alone. However, there was no consistency in the results. Combined treatments with X-rays did not provide any indication of a synergistic action between the RF fields and X-rays, either. Our data therefore do not support the hypothesis that RF fields act synergistically with chemical or physical mutagens. PMID- 11055796 TI - Effect of apolipoprotein E polymorphism and apolipoprotein A-1 gene promoter polymorphism on lipid parameters and premature coronary artery disease. AB - Genetic and environmental factors regulate lipid metabolism and phenotypic expression of CAD. In this study we assessed the effects of apoE gene polymorphism and apoA1 gene promoter polymorphism on lipid metabolism and risk for CAD. In a case-control study, 166 patients with CAD were compared with 130 healthy subjects. The apoE allele frequencies of patients vs. control group were 6.3% vs. 7.7% for e2, 84.3% vs. 84.6% for e3, and 9.4% vs. 7.7% for e4. Individuals with e3e4 and e4e4 genotypes had higher total (P = 0.023) and LDL cholesterol levels (P = 0.04) than individuals with other genotypes. There were no differences in lipid parameters between the subjects with the apoA1-GG genotype and subjects with AG or AA genotypes. However, univariate analysis revealed no association between risk genotypes (e3e4 and e4e4 genotypes) of apoE and CAD risk (OR = 1.1; 95% CI = 0.6-2.1, P = 0.8) as well as no association between the GG genotype and CAD risk (OR 0.7; 95% CI = 0.5-1.2, P = 0.19). No evidence for a synergistic interaction between e3e4 plus e4e4 genotypes and apoA1 GG genotype on CAD risk was found (OR = 1.3, 95% CI = 0.6-2.9; P = 0.5). One individual with familial defective apolipoprotein B-100 (Arg3500Gln) was found in each group. In conclusion, the apoE gene polymorphism affected the total and LDL cholesterol levels, whereas neither the apoE gene polymorphism nor the apoA-1 gene promoter polymorphism were shown to be independent risk factors for CAD in Slovenia. PMID- 11055797 TI - Gene aberrations in childhood brain tumors. AB - We present the results of the examination of prognostic markers in 40 children suffering from brain tumors. Prognostic markers such as amplification of the N myc and c-myc, deletion of the 17p, and DNA ploidy are indispensable factors for the determination of diagnosis. An increased number of c-myc gene copies was found in malignant brain tumors, especially embryonal, more often than reported in the literature. N-myc amplification occurs in our group seldom, but it seems to be a sign of worse prognosis in glial and embryonal brain tumors. DNA aneuploidy was not found very frequently, but in high-grade tumors only. PMID- 11055798 TI - Intratumoral IL-12 gene transfer improves the therapeutic efficacy of IL-12 but not IL-19. AB - We have compared the therapeutic activity of IL-12 and IL-18 in mice carrying IL 2 gene-transduced syngeneic sarcoma Mc12. The IL-2 gene-transduced sarcoma has previously been utilized as an irradiated, genetically modified tumour vaccine. Murine recombinant IL-12 was capable of suppressing growth of the IL-2 gene modified sarcoma Mc12 in syngeneic mice more efficiently than growth of the parental Mc12 sarcoma. In contrast, murine recombinant IL-18 could neither inhibit growth of the parental Mc12 sarcoma, nor suppress growth of its IL-2 gene modified transfectant. These results suggest that although both of these cytokines are functionally related and participate in the induction of IFN gamma production as well as in cell-mediated immune cytotoxicity, in the murine sarcoma system only IL-12 is therapeutically active and exerts its therapeutic effect in concert with the IL-2 gene. Thus, intratumoral IL-2 gene transfer improves the therapeutic efficacy of IL-12; administration of recombinant IL-12 should therefore be considered as adjuvant in IL-2 gene therapy with irradiated, genetically modified tumour vaccines. PMID- 11055799 TI - Human epidermal Langerhans cells are selectively recognized by galectin-3 but not by galectin-1. AB - Langerhans cells are dendritic antigen-presenting cells residing predominantly in the epidermis. Since endogenous galactoside-binding lectins with the jelly-roll motif (galectins) are known to trigger cellular responses, including mediator release, we investigated by lectin histochemistry the cells' capacity to bind two common members of this family, i.e. galectin-1 and -3. Actually, surrounding keratinocytes express a high level of galectin-3, and these cells can be considered as donors of this lectin to Langerhans cells. Employing biotinylated galectin-1 and -3, and concomitantly an antibody against CD1a as a second marker, to visualize the position of Langerhans cells in the human epidermis, the expression of galectin-3-reactive glycoligands in contrast to the lack of binding of galectin-1 was observed. Although the functional consequences of this selectivity are unclear, these results reveal an example for differential cellular reactivity towards two related endogenous lectins. PMID- 11055800 TI - Characterization of LK-1 monoclonal antibody against human sialophorin (CD43). PMID- 11055801 TI - Nonparametric regression sinogram smoothing using a roughness-penalized Poisson likelihood objective function. AB - We develop and investigate an approach to tomographic image reconstruction in which nonparametric regression using a roughness-penalized Poisson likelihood objective function is used to smooth each projection independently prior to reconstruction by unapodized filtered backprojection (FBP). As an added generalization, the roughness penalty is expressed in terms of a monotonic transform, known as the link function, of the projections. The approach is compared to shift-invariant projection filtering through the use of a Hanning window as well as to a related nonparametric regression approach that makes use of an objective function based on weighted least squares (WLS) rather than the Poisson likelihood. The approach is found to lead to improvements in resolution noise tradeoffs over the Hanning filter as well as over the WLS approach. We also investigate the resolution and noise effects of three different link functions: the identity, square root, and logarithm links. The choice of link function is found to influence the resolution uniformity and isotropy properties of the reconstructed images. In particular, in the case of an idealized imaging system with intrinsically uniform and isotropic resolution, the choice of a square root link function yields the desirable outcome of essentially uniform and isotropic resolution in reconstructed images, with noise performance still superior to that of the Hanning filter as well as that of the WLS approach. PMID- 11055802 TI - On segmenting the three-dimensional scan data of a human body. AB - Software for categorizing a cloud of more than 300,000 three-dimensional (3-D) surface data points, captured from a human subject, is presented. The software is part of an incremental approach that progressively refines the identification of human anthropometric landmarks. The first phase of identification is to orient and segment the human body data points. A step-by-step method for these tasks is presented. One of the algorithms, a discrete point cusp detector, plays a fundamental role in separating the data cloud. The theory and operation of the algorithm is explained. The discrete cusp detector can be used to separate any two cylindrical objects, in (3-D) data, whose boundaries touch. The software has been tested on over a hundred different body scan data sets and shown to be robust. PMID- 11055803 TI - Performance characteristics of the 3-D OSEM algorithm in the reconstruction of small animal PET images. Ordered-subsets expectation-maximixation. AB - Rat brain images acquired with a small animal positron emission tomography (PET) camera and reconstructed with the three-dimensional (3-D) ordered-subsets expectation-maximization (OSEM) algorithm with resolution recovery have better quality when the brain is imaged by itself than when inside the head with surrounding background activity. The purpose of this study was to characterize the dependence of this effect on the level of background activity, attenuation, and scatter. Monte Carlo simulations of the imaging system were performed. The coefficient of variation from replicate images, full-width at half-maximum (FWHM) from point sources and image profile fitting, and image contrast and uniformity were used to evaluate algorithm performance. A rat head with the typical levels of five and ten times the brain activity in the surrounding background requires additional iterations to achieve the same resolution as the brain-only case at a cost of 24% and 64% additional noise, respectively. For the same phantoms, object scatter reduced contrast by 3%-5%. However, attenuation degraded resolution by 0.2 mm and was responsible for up to 12% nonuniformity in the brain images suggesting that attenuation correction is useful. Given the effects of emission and attenuation distribution on both resolution and noise, simulations or phantom studies should be used for each imaging situation to select the appropriate number of OSEM iterations to achieve the desired resolution-noise levels. PMID- 11055804 TI - A fast implementation of the minimum spanning tree method for phase unwrapping. AB - A new implementation of the minimum spanning tree (MST) phase unwrapping method is presented. The time complexity of the MST method is reduced from O(n2) to O(n log2 n), where n is the number of pixels in the phase map. Typical 256 x 256 phase maps from magnetic resonance imaging can be unwrapped in seconds, compared with tens of minutes with the O(n2) implementation. This makes the pixel-level MST method time efficient and practically attractive. Index Terms-Image processing, magnetic resonance imaging, medical imaging, phase unwrapping. PMID- 11055805 TI - Image registration by maximization of combined mutual information and gradient information. AB - Mutual information has developed into an accurate measure for rigid and affine monomodality and multimodality image registration. The robustness of the measure is questionable, however. A possible reason for this is the absence of spatial information in the measure. The present paper proposes to include spatial information by combining mutual information with a term based on the image gradient of the images to be registered. The gradient term not only seeks to align locations of high gradient magnitude, but also aims for a similar orientation of the gradients at these locations. Results of combining both standard mutual information as well as a normalized measure are presented for rigid registration of three-dimensional clinical images [magnetic resonance (MR), computed tomography (CT), and positron emission tomography (PET)]. The results indicate that the combined measures yield a better registration function does mutual information or normalized mutual information per se. The registration functions are less sensitive to low sampling resolution, do not contain incorrect global maxima that are sometimes found in the mutual information function, and interpolation-induced local minima can be reduced. These characteristics yield the promise of more robust registration measures. The accuracy of the combined measures is similar to that of mutual information-based methods. PMID- 11055806 TI - Paraquat and iron-dependent lipid peroxidation. NADPH versus NADPH-generating systems. AB - The aim of this work was to study the effect of paraquat (P2+) on NADPH iron dependent lipid peroxidation (basal peroxidation) either in the presence of NADPH or in the presence of NADPH-generating systems. When NADPH is present, P2+ potentiates NADPH iron-dependent lipid peroxidation, but use of NADPH-generating systems cancels this effect. This may be attributed to certain components in NADPH-generating systems such as glucose-6-phosphate and sodium isocitrate, which act as iron chelators. The binding of iron by these molecules facilitates its reduction and enhances its reactivity toward dioxygen molecules, leading to the formation of reactive species capable of initiating lipid peroxidation, such as Fe3+-O2-*. Under these conditions of rapid basal peroxidation, any additional reduction of iron(III) by a reduced form of P2+ (P+*) has no apparent effect on the peroxidation itself, probably because the initial reaction between iron(II) and O2 followed by initiation of the peroxidation are both rate-limiting steps in the process. Consequently, any alteration of the composition of the reacting mixture (e.g., buffers or the generating system) must be taken into consideration because the formation of new iron chelates can change the rate of basal peroxidation and will modify the effect of redoxcycling molecules. PMID- 11055807 TI - A search for trace elements in some human intracranial tumors by instrumental neutron activation analysis. AB - A investigation was undertaken to measure the presence of trace elements in some intracranial tumors using the instrumental neutron activation analysis technique. The following 20 minor and trace elements were investigated: Na, Mg, Al, P, Cl, K, Ca, Cr, Mn, Fe, Co, Cu, Zn, As, Se, Br, Rb, Sb, I, and Cs. Our results are compared with other trace element analyses in human brain tissue. PMID- 11055808 TI - Correlations of calcium accumulations in arteries, veins, cartilages, ligaments, and bones in single humans. AB - To show the relationships of calcium accumulation in the thoracic aorta to the other tissues, calcium contents were determined with a microwave-induced plasma atomic emission spectrometer on arteries, veins, cartilages, ligaments, and bones. These tissues were resected from 18 individuals, consisting of 11 men and 7 women who died in the age range 59-91 yr. As thoracic and abdominal aortas are routinely used for radiographic examination of arterial calcification, they appear to be standard tissues of the calcium accumulation. The calcium accumulations were determined in the femoral artery, the superior and inferior venae cavae, the internal jugular vein, cartilages of the articular disk of the temporomandibular joint and the intervertebral disk, both the ligaments of the anterior cruciate ligament and the ligamentum capitis femoris, and the calcaneus, in contrast with the thoracic aorta. As calcium increased in the thoracic aorta, it increased in the femoral artery, the articular disk of the temporomandibular joint, the intervertebral disk, both ligaments of the anterior cruciate ligament, and the ligamentum capitis femoris, but it did not increase in veins, such as the superior and inferior venae cavae and the internal jugular vein. In contrast, it decreased in the calcaneus. PMID- 11055809 TI - Toxicity of cisplatin to the central nervous system of male rabbits. AB - The cytotoxic effects of platinum (Pt) were studied by intraparenchymal injection of 1 mg of cisplatin (CDDP) in male rabbits. Time-serial plasma Pt levels were used as CDDP clearance indices in brain and kidney tissues. The tissue samples were also examined histologically. Changes in the blood-brain barrier (BBB) were evaluated by horseradish peroxidase (HRP) extravasation. In the brain infusion group, Pt was detected in the plasma 30 min after the start of infusion. In the kidney, Pt was detected after 10 min of CDDP injection. The maximum plasma concentration of Pt in the brain group showed diffuse edema, neuronal necrosis, karyolysis, and HRP extravasation around the injection site. In contrast, the histological damage to kidneys was minimal. The results presented here show that direct infusion of CDDP caused the most extensive cytotoxicity in the brain. The low clearance rate of CDDP from the brain and BBB disruption may explain this behavior. PMID- 11055810 TI - Effect of peroxovanadate compound on phenylalanine hydroxylase gene expression. AB - Vanadium, a trace element in human cells and regarded as an essential nutrient, plays an active role in all tissues. It is known that peroxovanadate-nicotinic acid (POV), a complex compound of vanadium, can decrease hyperglycemia; however, its biochemical mechanism remains unclear. The object of the present study is to explore the hypoglycemia mechanism of POV at gene molecular levels. Rats rendered diabetic with streptozotocin were treated with POV. Total RNA was isolated from rat liver, and phenylalanine hydroxylase (PAH) mRNA abundance was determined by reverse transcriptase-polymerase chain reaction. PAH activity, blood glucose, and lipid levels were measured. Significantly increased hepatic PAH activity and corresponding mRNA with concomitant hyperglycemia and hyperlipemia were found in diabetic rats. These levels returned to normal after POV treatment and accompanied by negative glucosuria, normoglycemia, and normolipemia. The results from the current study indicates one of the mechanisms of POV action is to inhibit PAH gene expression and PAH activity, thus decreasing gluconeogenesis and hyperglycemia. At the same time, POV is able to promote diabetic recovery by lowering hyperlipemia. PMID- 11055811 TI - Magnetic resonance image and blood manganese concentration as indices for manganese content in the brain of rats. AB - Neurological disorders similar to parkinsonian syndrome and signal hyperintensity in brain on T1-weighted magnetic resonance (MR) images have been reported in patients receiving long-term total parenteral nutrition (TPN). These symptoms have been associated with manganese (Mn) depositions in brain. Although alterations of signal intensity on T1-weighted MR images in brain and of Mn concentration in blood are theoretically considered good indices for estimating Mn deposition in brain, precise correlations between these parameters have not been demonstrated as yet. Male Sprague-Dawley rats received TPN with 10-fold the clinical dose of the trace element preparation (TE-5) for 7 d. At 0, 2, 4, 6, and 8 wk post-TPN, the cortex, striatum, midbrain, and cerebellum were evaluated by MR images, and Mn concentration in blood and Mn content in these brain sites were measured by atomic absorption spectrometry. Immediately after TPN termination, signal hyperintensity in brain sites and elevated Mn content in blood and brain sites were observed. These values recovered at 4 wk post-TPN. A positive correlation was observed between either the signal intensity in certain brain sites or Mn content in blood and the relevant brain sites. Our observations suggest that the Mn concentration in blood and signal intensity in the brain sites on T1-weighted MR images are reliable indices for monitoring Mn contents in brain. PMID- 11055812 TI - A survey of trace elements in pteridophytes. AB - Concentration of 11 trace elements (Ca, Sc, Cr, Fe, Co, Zn, Rb, Cs, Ba, La, and Ce) in 96 pteridophytes (fern and fern ally species) was determined by instrumental neutron activation analysis to evaluate a concentration range for each element and also to find species characteristic in the uptake of trace elements. Asplenium trichomanes was found to accumulate Sc, Cr, and Co to the highest concentrations among 96 pteridophytes. The highest concentration of Ca and Zn was observed for Asplenium obscurum. The other Pteridophytes exhibited only one element whose concentration was the highest. A positive correlation was found between the concentrations of Fe and Sc, and also between the concentrations of Cr and Co. The remarkable accumulation of lanthanides (La and Ce) was observed mainly in diversifying genera (Polystichum and Dryopteris in Dryopteridaceae, Diplazium in Woodsiaceae, and Asplenium in Aspleniaceae). PMID- 11055813 TI - Blood lactate response and critical speed in swimmers aged 10-12 years of different standards. AB - It has previously been shown that measurement of the critical speed is a non invasive method of estimating the blood lactate response during exercise. However, its validity in children has yet to be demonstrated. The aims of this study were: (1) to verify if the critical speed determined in accordance with the protocol of Wakayoshi et al. is a non-invasive means of estimating the swimming speed equivalent to a blood lactate concentration of 4 mmol x l(-1) in children aged 10-12 years; and (2) to establish whether standard of performance has an effect on its determination. Sixteen swimmers were divided into two groups: beginners and trained. They initially completed a protocol for determination of speed equivalent to a blood lactate concentration of 4 mmol x l(-1). Later, during training sessions, maximum efforts were swum over distances of 50, 100 and 200 m for the calculation of the critical speed. The speeds equivalent to a blood lactate concentration of 4 mmol x l(-1) (beginners = 0.82 +/- 0.09 m x s(-1), trained = 1.19 +/- 0.11 m x s(-1); mean +/- s) were significantly faster than the critical speeds (beginners = 0.78 +/- 0.25 m x s(-1), trained = 1.08 +/- 0.04 m x s(-1)) in both groups. There was a high correlation between speed at a blood lactate concentration of 4 mmol x l(-1) and the critical speed for the beginners (r= 0.96, P < 0.001), but not for the trained group (r= 0.60, P> 0.05). The blood lactate concentration corresponding to the critical speed was 2.7 +/- 1.1 and 3.1 +/- 0.4 mmol x l(-1) for the beginners and trained group respectively. The percent difference between speed at a blood lactate concentration of 4 mmol x l( 1) and the critical speed was not significantly different between the two groups. At all distances studied, swimming performance was significantly faster in the trained group. Our results suggest that the critical speed underestimates swimming intensity corresponding to a blood lactate concentration of 4 mmol x l( 1) in children aged 10-12 years and that standard of performance does not affect the determination of the critical speed. PMID- 11055814 TI - State anxiety and motor performance: testing the conscious processing hypothesis. AB - Previous research has argued that skills acquired explicitly are more likely to fail under stressful conditions than skills that have been learned implicitly. The present study addressed an alternative explanation for the robustness under stress of implicit task performance. As implicit learners acquired the skill of golf putting while generating random letters, it is possible that they became desensitized to self-generated verbalizations and thus immune to the effects of competitive anxiety. We tested this interpretation while controlling for a further rival hypothesis generated by Eysenck's Processing Efficiency Theory. We also examined the effect of increased state anxiety on the kinematic processes underlying performance breakdowns. For task performance, we found evidence that partially supported the conscious processing hypothesis, while the results of the kinematic analysis of the putting stroke were equivocal. Analysis of self reported effort scores provided partial support for processing efficiency theory. PMID- 11055815 TI - Net forces during tethered simulation of underwater streamlined gliding and kicking techniques of the freestyle turn. AB - We assessed the net forces created when towing swimmers while gliding and kicking underwater to establish an appropriate speed for initiating underwater kicking, and the most effective gliding position and kicking technique to be applied after a turn. Sixteen experienced male swimmers of similar body shape were towed by a motorized winch and pulley system. A load cell measured net force (propulsive force - drag force) at speeds of 1.6, 1.9, 2.2, 2.5 and 3.1 m x s(-1). At each speed, the swimmers performed a prone streamline glide, a lateral streamline glide, a prone freestyle kick, a prone dolphin kick and a lateral dolphin kick. A two-way repeated-measures analysis of variance revealed significant differences between the gliding and kicking conditions at different speeds. The results demonstrated an optimal range of speeds (1.9 to 2.2 m x s(-1)) at which to begin underwater kicking to prevent energy loss from excessive active drag. No significant differences were found between the prone and lateral streamline glide positions or between the three underwater kicking techniques. Therefore, there appears to be no significant advantage in using one streamlining technique over another or in using one kicking style over another. PMID- 11055816 TI - Control of locomotion in expert gymnasts in the absence of vision. AB - The main aim of this study was to determine how gymnasts are affected by the removal of vision when executing simple moves. A secondary aim was to establish whether crucial sensory cues exist for blindfolded gymnasts. Eight expert gymnasts were asked to maintain a straight displacement during three types of blindfolded locomotion: walking, steering a wheelchair and verbally ordering a second person pushing their wheelchair. In the first two conditions, active displacement made proprioceptive cues available to update the body trajectory. In the last condition, however, proprioceptive cues were greatly reduced, since the gymnasts displaced passively. The performance of the gymnasts was compared to that of eight experts in other non-gymnastic sports (control group). The results showed that the participants veered in all conditions. However, except in the verbal condition, the gymnasts departed less from linearity than the controls. We conclude that: (1) even for a simple motor task, gymnasts' performance is altered by eliminating vision; (2) compared with other sportsmen and women, gymnasts are better able to deal with the absence of vision when proprioceptive cues are available. These findings suggest two possible explanations: (1) gymnasts are more able to 'pick up' crucial information and (2) a gymnast's proprioceptive system is more sensitive. PMID- 11055817 TI - Hierarchical confirmatory factor analysis of the Flow State Scale in exercise. AB - In this study, we examined the factor structure and internal consistency of the Flow State Scale using responses of exercise participants. This self-report questionnaire consists of nine subscales designed to assess flow in sport and physical activity. It was administered to 1231 aerobic dance exercise participants. Confirmatory factor analyses were used to test three competing measurement models of the flow construct: a single-factor model, a nine-factor model and a hierarchical model positing a higher-order flow factor to explain the intercorrelations between the nine first-order factors. The single-factor model showed a poor fit to the data. The nine-factor model and the hierarchical model did not show an adequate fit to the data. All subscales of the Flow State Scale displayed acceptable internal consistency (alpha > 0.70), with the exception of transformation of time (alpha = 0.65). Collectively, the present results do not provide support for the tenability of the single-factor, nine-factor or hierarchical measurement models in an exercise setting. PMID- 11055818 TI - Effects of bicycle frame ergonomics on triathlon 10-km running performance. AB - It is perceived that, during the triathlon or duathlon, cycling with a steep (> 76 degrees) rather than a shallow (< 76 degrees ) frame geometry might attenuate the fatigue associated with progression from the cycle to run disciplines and improve subsequent 10-km running performance. This is based on anecdotal testimony from athletes purporting to have experienced improved performance; no empirical evidence exists. To evaluate this view, eight male triathletes completed a counterbalanced, 40-km cycle ride at two frame geometries (73 degrees and 81 degrees) at approximately 70% VO2peak. Immediately after completion of each 40-km cycle, a self-paced 10-km treadmill time trial was undertaken, during which physiological, kinematic and performance variables were measured. The 10-km run performance (mean +/- s: 42:55 +/- 4:19 vs 46:15 +/- 4:52 min; P< 0.01) and combined cycle and run performance (1:45:49 +/- 5:45 vs 1:50:33 +/- 6:08; P< 0.001) were faster in the 81 degrees than the 73 degrees condition. Improvements in performance were most prominent during the first 5 km of the run (21:41 +/- 2:15 vs 24:15 +/- 2:31 min in the 81 degrees and 73 degrees conditions respectively). These improvements were not evident during the second 5 km of the run. No differences in physiological variables were noted, although heart rate, stride length and stride frequency were increased during the 81 degrees condition (P < 0.05). Modifying frame geometry from a seat tube angle of 73 degrees to 81 degrees improves 10-km running and combined cycle plus run performance. These improvements in performance might relate to alterations during the cycling phase, which minimizes the 'residual effect' of this (i.e. the adverse changes in substrate availability, thermoregulatory, cardiovascular and biomechanical factors felt immediately after transition from cycling to running) and attenuates negative changes in physiological and kinematic responses during the 10-km run. PMID- 11055819 TI - Energy conversion rates during sprinting with an emphasis on the performance of female athletes. AB - Computed results from a mathematical model of the bioenergetics of sprinting, which incorporates a three-equation representation of anaerobic metabolism, were compared with measured distance-time data for female athletes from the finals of the 100-m event at the World Championships of 1987. The computed results closely model the performance of the competitors over the course of the entire race. The three main contributions to anaerobic metabolism were investigated and comparisons were made between male and female sprinters. Whereas the time constants for adenosine triphosphate (ATP) and phosphocreatine utilization for the two sexes were found to be similar, for oxygen-independent glycolysis the time constants of the female athletes were found to be higher. The maximum powers generated by female athletes during ATP conversion and glycolysis were only slightly lower than the figures found for male athletes, but the value for phosphocreatine utilization was substantially lower. The lower value for phosphocreatine utilization might explain the more pronounced fall-off in running speed over the latter stages of a race that female athletes experience in comparison with men. Although anaerobic sources dominate energy provision for both male and female sprinters, the calculations show that the latter make greater use of aerobic energy supplies. PMID- 11055820 TI - COX-2-Specific inhibitors--the emergence of a new class of analgesic and anti inflammatory drugs. AB - The prostaglandin series of bioactive compounds is formed by the interaction of two distinct but related enzymes, cyclo-oxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). COX-1 is a constitutive form which is present mainly in the gastric mucosa, kidney and platelets. COX-2 is mainly an inducible form, although also to some extent present constitutively in the CNS, the juxtaglomerular apparatus of the kidney and in the placenta during late gestation. Both isoforms contribute to the inflammatory process, but COX-2 is of considerable therapeutic interest as it is induced, resulting in an enhanced formation of prostaglandins, during acute as well as chronic inflammation. Conventional NSAIDs inhibit both isoforms to a similar extent and in an approximately equal dose and concentration range. The two recently developed and clinically available selective COX-2 inhibitors, celecoxib and rofecoxib, are about 100-1000 times more selective on the COX-2 than on the COX-1 isoform. In Europe rofecoxib is today indicated for the symptoms and signs of osteoarthritis, whereas celecoxib is indicated for both osteoarthritis and rheumatoid arthritis. The major clinical interest of these drugs has been related to the lower incidence of gastrointestinal bleeding which, with the conventional COX-1/COX-2 agents has been a source of hospitalisation, disablement and death, especially in the elderly. Clinical trials have convincingly demonstrated that celecoxib and rofecoxib in clinical use induce very few gastrointestinal complications compared to conventional and non selective NSAIDs. However, the well known contraindications for NSAIDs, such as late pregnancy, aspirin-induced asthma, congestive heart failure and renal dysfunction, will so far apply also to the COX-2 inhibitors. Compared to the traditional and non-selective NSAIDs, COX-2 inhibitors may provide an insight into additional therapeutic areas, such as gastrointestinal cancer and dementia, where the potential relevance to COX-2 mechanisms are currently being explored and clinical trials being performed. With the rapid clinical acceptance of celecoxib and rofecoxib, knowledge about their clinical usefulness in various inflammatory disease states and pain disorders is increasing. For the many patients suffering from such conditions, the selective COX-2 inhibitors are likely to become a significant addition to the therapeutic arsenal of analgesic and anti-inflammatory drugs. PMID- 11055821 TI - Haematogenous vertebral osteomyelitis in the elderly. AB - The aim of our study was to analyse the characteristics of haematogenous vertebral osteomyelitis (HVO) in the elderly. A retrospective comparative analysis of the medical records of 72 patients (38 younger than 63 years, group 1, and 34 aged 63 years and over, group 2) with haematogenous vertebral osteomyelitis of confirmed aetiology was carried out. Intravenous drug addiction and infection with the human immunodeficiency virus were seen in 4/38 (10%) and 5/38 (13%) patients from group 1 and 0/34 patients (0%) from group 2 (P = 0.05 and 0.035, respectively). Seven of 34 elderly (20%) and 0/38 (0%) young individuals had recently had surgery (P = 0.0036). Escherichia coli was isolated in 7/34 elderly (20%) and 0/38 (0%) young patients (P = 0.0036). The remaining studied data did not reach statistical significance. Recent surgery is a risk factor for developing HVO in the elderly, the urinary tract being the source of the pathogen in a large number of elderly patients with spinal infection. PMID- 11055822 TI - Seronegative spondyloarthropathy initiated by physical trauma. AB - We undertook this study to demonstrate the pattern of onset and the course of arthritis on the traumatised joint in spondyloarthropathy (SpA) initiated by physical trauma. Among 288 patients with SpA, 12 (4.2%) whose arthropathies were associated with trauma were reviewed retrospectively. There were seven patients with ankylosing spondylitis (AS), three with juvenile onset AS and two undifferentiated SpA. The type of trauma was direct injury to the joint and injuries at other sites, except in spinal surgery, for example. In eight cases the initial evidence of disease was peripheral arthritis. The disease first occurred in traumatised joints in five cases. Only three cases showed recurrent inflammatory episodes in the traumatised joints throughout the disease course. SpA initiated by trauma initially manifested as peripheral arthritis at the traumatised joints in about half of the cases. Inflammatory episodes preferentially involved other joints apart from the traumatised joints throughout the whole course of the disease. PMID- 11055823 TI - Elevation of serum soluble tumour necrosis factor receptors in patients with polymyositis and dermatomyositis. AB - The aim of the study was, to examine the relationship between serum levels of soluble tumour necrosis factor receptors (sTNF-R) and the gene expression of two types of receptor for TNF (TNF-R), a 55 a receptor (TNF-R1) and a 75 kDa receptor (TNF-R2), bloodin peripheral mononuclear cells (PBMC) from patients with polymyositis and dermatomyositis (PM/DM). Soluble tumour necrosis factor receptor 1 (sTNF-R1) and soluble tumour necrosis factor receptor 2 (sTNF-R2) levels in sera from patients were measured by enzyme-linked immunosorbent assay. Expression of TNF-R1 and TNF-R2 mRNAs in PBMC was analysed by Northern blotting. Serum sTNF R1 and sTNF-R2 levels were elevated significantly in 25 patients with active stage PM/DM, compared to those in 18 patients with inactive-stage PM/DM and 32 normal controls. Serum concentrations of sTNF-R1 and sTNF-R2 were correlated with PM/DM disease activity. TNF-R1 gene expression was enhanced in freshly isolated PBMC from patients with active-stage PM/DM. In contrast, TNF-R2 mRNA was expressed constitutively in patients with active-stage PM/DM and in normal controls. The expression of TNF-R1 and TNF-R2 mRNAs in PBMC and elevation of their soluble forms in the sera of patients with active-stage PM/DM suggest increased proteolytic cleavage of cell surface TNF-R from PBMC in patients with active-stage PM/DM, and that sTNF-R may regulate TNF-alpha-mediated muscle fibre damage in PM/DM. PMID- 11055824 TI - Elevated nitric oxide production in patients with primary Sjogren's syndrome. AB - Nitric oxide (NO) production is elevated in patients with inflammatory disorders. We have previously shown increased NO production in patients with rheumatoid arthritis and systemic lupus erythematosus. In this study we used nitrite and citrulline levels as surrogate markers of NO production in patients with primary Sjogren's syndrome (SS) and measured their levels by spectrophotometry. Fifteen patients and 15 age- and sex-matched controls were studied. Mean nitrite levels in patients were 582.3+/-208.3 nmol/ml, but those in controls were significantly lower, at 203.2-106.9 nmol/ml (p<0.001). Citrulline levels were 2820.4+/-933.9 nmol/ml in patients and were significantly higher than 217.4+/-144.8 nmol/ml, the levels in controls (p<0.0001). Mean levels of both nitrite and citrulline were significantly higher in patients with arthritis than in those who had no joint manifestations (p<0.05). There was no correlation between NO production and other variables, such as age, disease duration, drug therapy and antinuclear antibodies or rheumatoid factor positivity. Increased NO production may be partly a reflection of the presence of arthritis in five patients. It is concluded that there is increased NO production in patients with primary SS, especially if they have associated arthritis. PMID- 11055825 TI - Significance of low mRNA levels of interleukin-4 and -10 in mononuclear cells of the synovial fluid of patients with rheumatoid arthritis. AB - Our objective was to investigate the clinical significance of Th1 and Th2-type cytokines, such as interferon-gamma (IFNgamma) interleukin (IL)-12, IL-4 and IL 10, in the mononuclear cells (MNC) of the synovial fluid (SF) in patients with rheumatoid arthritis (RA). The cytokine production in the MNC obtained from the SF (SF-MNC) in 30 patients with RA and 10 with gout was examined by measuring the mRNA levels of IFNgamma, IL-12, IL-4 and IL-10 by semiquantitative RT-PCR. The mRNA levels of IFNgamma, IL-4 and IL-10 were significantly higher in the SF-MNC of RA patients than in those of gout patients (p<0.001, p<0.01 and p<0.001, respectively). Correlations between mRNA levels were significant for IL-12 and IL 4, IL-12 and IL-10, and IL-4 and IL-10 (p<0.05). The mRNA levels of IL-4 and IL 10 were very low compared to those of IL-12 in seven of the 30 patients with RA; all of these patients were in stage 4, and serum levels of CRP, ESR and blood platelet count which are considered as indices of the severity of inflammation, were significantly elevated in these seven patients compared to the other 23 RA patients. The markedly reduced synthesis of both IL-4 and IL-10 mRNA could be considered to be related to the progression and/or activity of RA. The results of this study therefore indicate an imbalance in the levels of Th1 and Th2 cytokines at the site of inflammation in RA, and draw attention to the possibility of treatment of progressive or intractable RA with IL-4 and/or IL-10. PMID- 11055826 TI - Clinical and serological aspects of patients with anti-Jo-1 antibodies--an evolving spectrum of disease manifestations. AB - The aim of this study was to compare ELISA, immunodiffusion and immunoblot for the detection of anti-Jo-1 antibodies, and to investigate the association of the results with clinical manifestations. In two medical centres for rheumatology and one for pulmonology, all patients with suspected connective tissue disease were screened over a 5-year period for anti-Jo-1 antibodies by ELISA. Positive sera were controlled in another laboratory by immunodiffusion. If immunodiffusion was negative, sera were controlled again by ELISA. ELISA-positive immunodiffusion negative sera were tested by immunoblotting. The patients were characterised clinically, and their clinical signs and symptoms were compared with those of 257 patients with anti-Jo-1 antibodies published in 15 case series and 30 case reports. Twenty-five patients had a positive ELISA test. Fifteen sera were positive by ELISA and immunodiffusion (group 1). Three sera showed high titres in both ELISA tests with negative immunodiffusion and immunoblot (group 2). Seven sera showed low titres in both ELISA tests. The results were negative in the other tests (group 3). Patients in groups 1 and 2 could be classified as Jo-1 syndrome patients. Of these 18 patients, 15 had arthritis, 14 had myositis and 14 had interstitial lung disease. Only four patients had myositis at disease onset. We describe four unusual patients with Jo-1 syndrome in detail: 1. Long history of seronegative rheumatoid arthritis; 2. Sjogren's syndrome with Ro- and La antibodies; 3. Scleroderma and bronchial carcinoma with centromere antibodies; 4. Corticoid-sensitive psychosis. Patients with suspected connective tissue disease may be screened for anti-Jo-1 antibodies by ELISA. It detects some patients that are missed by immunodiffusion. Especially lower ELISA titres should be controlled by another method because of the low specificity of the test. The clinical picture is variable. Most patients have features other than myositis at disease onset. PMID- 11055827 TI - Systemic sclerosis: another rheumatic disease associated with hepatitis C virus infection. AB - The first case of a patient with chronic infection with hepatitis C virus who developed systemic sclerosis, manifested by severe Raynaud's phenomenon, progressive skin thickening, painful fingertip ulcers, dysphagia and Sjogren's syndrome, is described. The role of interferon therapy is discussed. PMID- 11055828 TI - Mixed connective tissue disease associated with skin defects of livedoid vasculitis. AB - A 21-year-old woman who had a 2-year history of mixed connective tissue disease (MCTD) developed rapidly evolving ulcers consistent with livedoid vasculitis (LV) in all distal extremities. She presented clinically with Raynaud's phenomenon, polyarthritis and swollen hands; serologically with high titres of ANA and anti nRNP; and immunogenetically with HLA-DR4 and HLA-DR53. Although there was initial success in treatment except for the skin defects over the ankles, the patient died from disseminated intravascular coagulation. We suggest that LV may be a poor prognostic manifestation in MCTD. PMID- 11055829 TI - Synchronous pyomyositis and septic hip arthritis. AB - The authors report a rare concomitant pyogenic infection of the iliopsoas, iliacus and external obturator muscles and of the hip joint in a 68-year-old woman. Because the patient showed the classic symptomatic triad of limping, hip pain and fever, in addition to positive hip arthrocentesis, the diagnosis of septic hip arthritis was routine, but the simultaneous pyomyositis was almost overlooked. Unusual localised heat and swelling on the front of the proximal thigh prompted a CT scan that identified remarkable muscle abscesses in addition to the septic arthritis. Surgical debridement and antibiotics resolved the infection relatively rapidly without sequelae. We noted that reaching a definitive diagnosis of such a concomitant infection requires a suspicion of the presence of pyomyositis, which can be definitively determined using advanced imaging studies. PMID- 11055830 TI - Urticaria as a presenting manifestation of adult-onset Still's disease. AB - Adult-onset Still's disease (AOSD) is a rare disorder of unknown aetiology, characterised by high spiking fever, an evanescent, erythematous, maculopapular rash, arthralgia or arthritis, lymphadenopathy, hepatosplenomegaly, sore throat and serositis. It is associated with marked leukocytosis, high erythrocyte sedimentation rate, increased level of serum ferritin and negative rheumatoid factor and antinuclear antibody tests. Here we report a patient in whom an urticaria-like rash was an uncommon presenting clinical feature of AOSD. To our knowledge, this association has only been reported once before. PMID- 11055831 TI - Pulmonary nodule and aggressive tibialis posterior tenosynovitis in early rheumatoid arthritis. AB - We report the case of a 34-year-old man with a rheumatoid pulmonary nodule preceding the development of articular symptoms of rheumatoid arthritis. Pulmonary nodules are a well known feature of rheumatoid arthritis and are mostly seen in severe established rheumatoid factor-positive cases. To differentiate between benign and malign pulmonary nodules we discuss the use of positron emission tomography (PET). Despite intensive therapy with steroids and methotrexate in our patient, within months he developed a severe tibialis posterior tendinitis, with partial rupture and evolution to a planovalgus deformity requiring surgery. Both these symptoms are rare but demonstrate the need for close follow-up in early rheumatoid arthritis. PMID- 11055832 TI - Oral prednisolone improved acetylcholine-induced sweating in Sjogren's syndrome related anhidrosis. PMID- 11055833 TI - Nails 'obstructing' finger growth in length in primary hypertrophic osteoarthropathy (PHO). PMID- 11055834 TI - Sacral insufficiency fractures. High association with pubic rami fractures. PMID- 11055835 TI - Preventing pneumococcal disease among infants and young children. Recommendations of the Advisory Committee on Immunization Practices (ACIP). AB - In February 2000, a 7-valent pneumococcal polysaccharide-protein conjugate vaccine (Prevnar, marketed by Wyeth Lederle Vaccines) was licensed for use among infants and young children. CDC's Advisory Committee on Immunization Practices (ACIP) recommends that the vaccine be used for all children aged 2-23 months and for children aged 24-59 months who are at increased risk for pneumococcal disease (e.g., children with sickle cell disease, human immunodeficiency virus infection, and other immunocompromising or chronic medical conditions). ACIP also recommends that the vaccine be considered for all other children aged 24-59 months, with priority given to a) children aged 24-35 months, b) children who are of Alaska Native, American Indian, and African-American descent, and c) children who attend group day care centers. This report includes ACIP's recommended vaccination schedule for infants at ages 2, 4, 6, and 12-15 months. This report also includes a pneumococcal vaccination schedule for infants and young children who are beginning their vaccination series at an older age and for those who missed doses. In addition, this report updates earlier recommendations for use of 23 valent pneumococcal polysaccharide vaccine among children aged > or =2 years. Among children aged 24-59 months for whom polysaccharide vaccine is already recommended, ACIP recommends vaccination with the new conjugate vaccine followed, > or =2 months later, by 23-valent polysaccharide vaccine. Conjugate vaccine has not been studied sufficiently among older children or adults to make recommendations for its use among persons aged > or =5 years. Persons aged > or =5 years who are at increased risk for serious pneumococcal disease should continue to receive 23-valent polysaccharide vaccine in accordance with previous ACIP recommendations. PMID- 11055836 TI - Chemoprevention of breast cancer by tamoxifen: risks and opportunities. AB - The antiestrogen tamoxifen is widely used in the adjuvant therapy of breast cancers in women and helps to prevent the occurrence of breast tumors in healthy women. However, epidemiological studies have shown tamoxifen treatment to be associated with a 2- to 5-fold increased risk of endometrial cancer. In rats but not in mice, long-term administration of tamoxifen results in an increase in hepatocellular carcinomas. Mechanistically, this occurs through metabolic activation of the drug, mainly by the CYP3A family, to an electrophilic species, that causes DNA damage in target tissues, and subsequently leads to gene mutations. It is controversial whether low levels of DNA damage occur in human uterine tissues, and there is no evidence that this can be causally related to the mechanisms of carcinogenesis. In healthy women, the risk:benefits for the use of tamoxifen is in part related to the risk of developing breast cancer. The results from the carcinogenicity studies in rats do not predict the likelihood that women will develop liver cancer or indeed cancers in other organs. The mechanism of endometrial cancer in women remains unresolved, but the experience with tamoxifen has highlighted the potential problems that need to be addressed in the assessment of future generations of selective estrogen receptor modulators. PMID- 11055837 TI - Carcinogenicity and genotoxicity of ethylene oxide: new aspects and recent advances. AB - Long-term inhalation studies in rodents have presented unequivocal evidence of experimental carcinogenicity of ethylene oxide, based on the formation of malignant tumors at multiple sites. However, despite a considerable body of epidemiological data only limited evidence has been obtained of its carcinogenicity in humans. Ethylene oxide is not only an important exogenous toxicant, but it is also formed from ethylene as a biological precursor. Ethylene is a normal body constituent; its endogenous formation is evidenced by exhalation in rats and in humans. Consequently, ethylene oxide must also be regarded as a physiological compound. The most abundant DNA adduct of ethylene oxide is 7-(2 hydroxyethyl)guanine (HOEtG). Open questions are the nature and role of tissue specific factors in ethylene oxide carcinogenesis and the physiological and quantitative role of DNA repair mechanisms. The detection of remarkable individual differences in the susceptibility of humans has promoted research into genetic factors that influence the metabolism of ethylene oxide. With this background it appears that current PBPK models for trans-species extrapolation of ethylene oxide toxicity need to be refined further. For a cancer risk assessment at low levels of DNA damage, exposure-related adducts must be discussed in relation to background DNA damage as well as to inter- and intraindividual variability. In rats, subacute ethylene oxide exposures on the order of 1 ppm (1.83 mg/m3) cause DNA adduct levels (HOEtG) of the same magnitude as produced by endogenous ethylene oxide. Based on very recent studies the endogenous background levels of HOEtG in DNA of humans are comparable to those that are produced in rodents by repetitive exogenous ethylene oxide exposures of about 10 ppm (18.3 mg/m3). Experimentally, ethylene oxide has revealed only weak mutagenic effects in vivo, which are confined to higher doses. It has been concluded that long-term human occupational exposure to low airborne concentrations to ethylene oxide, at or below current occupational exposure limits of 1 ppm (1.83 mg/m3), would not produce unacceptable increased genotoxic risks. However, critical questions remain that need further discussions relating to the coherence of animal and human data of experimental data in vitro vs. in vivo and to species-specific dynamics of DNA lesions. PMID- 11055838 TI - Molecular mechanisms of apoptosis induced by cytotoxic chemicals. AB - The purpose of this review article is to discuss established molecular mechanisms of apoptosis and their relevance to cell death induced by environmental toxicants. Apoptosis is a highly regulated form of cell death distinguished by the activation of a family of cysteine-aspartate proteases (caspases) that cleave various proteins resulting in morphological and biochemical changes characteristic of this form of cell death. Abundant evidence supports a role for mitochondria in regulating apoptosis. Specifically, it seems that a number of death stimuli target these organelles and stimulate, by an unknown mechanism, the release of several proteins, including cytochrome c. Once released into the cytosol, cytochrome c binds to its adaptor molecule, Apaf-1, which oligomerizes and then activates pro-caspase-9. Caspase-9 can signal downstream and activate pro-caspase-3 and -7. The release of cytochrome c can be influenced by different Bcl-2 family member proteins, including, but not limited to, Bax, Bid, Bcl-2, and Bcl-X(L). Bax and Bid potentiate cytochrome c release, whereas Bcl-2 and Bcl-X(L) antagonize this event. Although toxicologists have traditionally associated cell death with necrosis, emerging evidence suggests that different types of environmental contaminants exert their toxicity, at least in part, by triggering apoptosis. The mechanism responsible for eliciting the pro-apoptotic effect of a given chemical is often unknown, although in many instances mitochondria appear to be key participants. This review describes our current understanding of the role of apoptosis in environmental toxicant-induced cell death, using dioxin, metals (cadmium and methylmercury), organotin compounds, dithiocarbamates, and benzene as specific examples. Finally, we conclude with a critical discussion of the current knowledge in this area and provide recommendations for future directions. PMID- 11055839 TI - Albumen height and yolk and embryo compositions in broiler hatching eggs during incubation. AB - The relationship of albumen height (AH) to the compositions of yolks and embryos in hatching eggs from a young (30 wk of age) broiler breeder flock was evaluated during incubation. On Day 2 of incubation, egg weight, yolk weight, and yolk moisture, lipid, and fatty acid contents were determined in eggs from broiler breeders previously identified as laying eggs of either low or high AH. In addition, egg weight, wet and dry embryo weight, and embryo moisture and protein contents were determined on Days 10, 12, and 16, and embryo lipid content was determined on Days 12 and 16. Yolk and embryo weights were expressed as percentages of sampled egg weight. Egg, yolk, and wet embryo weights, yolk moisture and lipid contents, and embryo moisture, protein, and lipid contents were not affected by AH; however, yolk myristic acid concentration was higher, and yolk linoleic acid concentration was lower, in low AH eggs on Day 2 of incubation. Furthermore, on Day 16, dry embryo weight was significantly higher in low AH eggs. Young breeder hens laying eggs of different AH may also produce egg yolks with different fatty acid compositions. Differences in yolk fatty acid profiles between AH groups during early incubation may impact subsequent embryo DM weight without associated effects on embryo moisture, protein, or lipid contents. PMID- 11055840 TI - The toxicity of purified fumonisin B1 in broiler chicks. AB - An investigation of the toxicity of fumonisin B1 (FB1), a toxic metabolite of Fusarium moniliforme, in broiler chicks was conducted. Purified FB1 (98.1% pure) was incorporated into the diets of broiler chicks at 0, 20, 40, and 80 mg/kg, and fed to chicks from 0 to 21 d of age. Dietary FB1, at concentrations of 80 mg/kg or less, did not adversely affect body weight, feed efficiency, or water consumption of broiler chicks. The relative weights of the liver, spleen, kidney, proventriculus, and bursa of Fabricius were also unaffected (P < 0.05) by any dietary concentration of FB1 compared with the control (0 mg/kg) group. Total liver lipids of chicks fed 40 or 80 mg FB1/kg were significantly lower than those of the chicks fed either 0 or 20 mg FB1/kg of feed. Liver sphinganine concentration and the sphinganine:sphingosine ratio were increased significantly in all treated groups. Chicks fed dietary FB1 at 80 mg/kg had significantly higher serum glutamate oxaloacetate aminotransaminase:aspartate aminotransferase ratios and levels of free sphinganine in the serum. The results of this investigation agree with the results previously described, in which FB1 was supplied to diets from the use of F. moniliforme-contaminated grain; therefore, the use of such material as the source of the mycotoxin in animal feeding studies is appropriate. PMID- 11055841 TI - Effects of relative humidity during the last five days of incubation and brooding temperature on performance of broiler chicks from young broiler breeders. AB - Broiler hatching eggs were subjected to one RH condition (53% RH) from 0 to 16 d and switched to one of three different RH conditions (43, 53, or 63%) from transfer at 16 d to pull time at 21.67 d of incubation. The broiler breeder ages were 27, 29, and 31 wk for Trial 1, and 26, 28, and 30 wk for Trial 2 in Experiment 1. For both trials, BW at hatch, BW at pull, hatch time, and chick weight loss between hatching and pull were measured. All unhatched eggs were opened and examined macroscopically. Experiment 2 was conducted using the chicks produced in Experiment 1. The chicks were immediately placed after pull and randomly distributed by sex into brooding pens by RH treatment in Trial 1, and by sex, RH, and brooding temperature in Trial 2. Two brooding conditions, warm and cool, were used in Trial 2. Body weight at placement, BW gain to 12 d, feed conversion, and mortality were recorded. There was no effect of RH treatment on fertile hatchability. Body weight at hatch, pull, and placement increased with hen age. Mortality was higher for the late-hatching chicks in the warm brooding conditions in Trials 1 and 2 of Experiment 2, whereas mortality was significantly greater for the early-hatching chicks and those chicks hatched at 43% RH under the cool-brooding temperature conditions. Relative humidity treatment had no effect on chick performance in the presence of warm brooding conditions. Warm brooding conditions improved BW gain to 12 d compared with cold brooding conditions. Overall, optimal chick performance was achieved at 53% RH. Chick quality problems that may be due to high or low RH during incubation can be ameliorated by proper brooding conditions. PMID- 11055842 TI - Effect of delayed placement on the incidence of Campylobacter jejuni in broiler chickens. AB - An experiment was conducted with broiler chickens to evaluate the effect of delayed placement on reused litter and the isolation of Campylobacter jejuni. The experiment also assessed the presence of C. jejuni in the crop following feed withdrawal periods in cages vs floor environments. Trial 1 utilized 320 female broiler chicks obtained from a commercial hatchery. The chicks were randomly placed into the following experimental groups that were replicated four times with 20 chicks per pen: 1) 0-h, 2) 24-h, 3) 48-h, and 4) 72-h delayed placement. Fecal samples were collected via the cloaca at 7, 14, 21, and 28 d of age; enriched in Bolton broth; and plated onto BBL agar. Campylobacter jejuni was isolated at 1 wk of age in the 48- and 72-h experimental groups but did not differ significantly from the others. Sampling results at 14 d of age showed that 63, 68, 73, and 80% of chicks were positive for the 0-, 24-, 48-, and 72-h treatments, respectively. At 28 d of age, 100% of all chicks sampled tested positive. In Trial 2, 60 market age broilers were evaluated for the presence of C. jejuni in the crop by subjecting them to 0-, 4-, 8-, 12-, and 16-h feed withdrawal times on litter or in wire cages. Crops were collected aseptically from the broilers, stomached, enriched in Bolton broth, and then plated on BBL agar. There were no significant (P < 0.05) differences between cage and floor withdrawal times for the presence of C. jejuni in the crop samples of broilers; however, there were significant differences in the length of withdrawal on the presence of C. jejuni. Collectively, these results suggested that the isolation of C. jejuni occurred earlier in broilers that were subjected to delayed placement on reused litter and that extended feed withdrawal times in cages or on litter may increase the possibility that the crop of broilers may contain a higher isolation rate of C. jejuni. PMID- 11055843 TI - The effect of air velocity on broiler performance and feed and water consumption. AB - Two trials were conducted to determine the effect of air velocity on feed and water consumption at a constant temperature of 27 C and a daily cyclic temperature of 22-32-22 C. Air velocity over the broilers was <15 or 120 m/min. These temperature and air velocity treatments were arranged in a 2 x 2 factorial design in eight environmental chambers, with two replications of each treatment. The air velocity treatments were applied, and total feed and water consumption and daily patterns of consumption were determined for broilers from 21 to 49 d of age. Broilers exposed to the high air velocity consumed less water and more feed, gained more weight, and had an improved feed:gain ratio. The high air velocity had little effect on daily patterns of feed and water consumption. Both feed and water consumption were depressed during the peak of the daily cyclic temperature. PMID- 11055844 TI - The effect of early age feed restriction on subsequent response to high environmental temperatures in female broiler chickens. AB - This study was conducted to determine whether early age feed restriction improves heat tolerance in female broiler chickens. Chicks were brooded for 3 wk and then maintained at 24+/-1 C. On Day 0, chicks were assigned to one of four feeding regimens; each regimen was applied to four cages of chicks. The feeding regimens were 1) ad libitum feeding (ALF); 2) 40% feed restriction at 4, 5, and 6 d of age (F40); 3) 60% feed restriction at 4, 5, and 6 d of age (F60); and (4) 80% feed restriction at 4, 5, and 6 d of age (F80). From 35 to 41 d of age, all birds were exposed to 38+/-1 C for 2 h/d. Serum concentrations of glucose were elevated by the heat challenge, but were not affected by the feeding regimen. The heat treatment resulted in hypocholesteremia among ALF and F80 chicks, whereas the concentrations increased and remained constant in the F60 and F40 birds, respectively. Subjecting chicks to F60 improved growth and survivability and reduced heterophil to lymphocyte ratios (H/L) in response to the heat treatment as compared with the ALF and F80 regimens. The survivability rate and H/L of F40 chicks were similar to those attained by chicks on other regimens. Newcastle disease antibody titer of ALF birds declined with duration of heat treatment. It is concluded that the F60 regimen is beneficial for alleviating, at least in part, the detrimental effects of heat stress in female broiler chickens. PMID- 11055845 TI - Elimination of early Salmonella enteritidis infection after treatment with competitive-exclusion culture and enrofloxacin in experimentally infected chicks. AB - The effect of normal avian gut flora (NAGF) and enrofloxacin administration on the early infection of young chicks by Salmonella enteritidis (SE) was determined using day-old White Leghorn chicks. Day- old chicks were divided into two groups, untreated control and NAGF-treated, and then infected with 10(6) cfu of SE per chick by oral gavage. The untreated, infected chicks were further divided into two groups and were either left untreated or medicated with a regimen of 10 mg/kg of enrofloxacin in drinking water daily for 10 d, followed by two doses of NAGF beginning at 10 and 8 wk of age in Trial 1 and Trial 2, respectively. Liver, spleen, and cecum samples were tested for the presence of SE, and immunological responsiveness was investigated up to 12 wk of age. Compared with the untreated group, the cecal colonization of SE was significantly (P < 0.05) decreased in the NAGF-treated group in Trials 1 and 2. No significant differences in organ infection were observed in the NAGF-treated vs. untreated birds. Although a significant effect of the combined treatment of enrofloxacin treatment and NAGF on the early infection was not shown in Trial 1, compared with enrofloxacin only or the untreated group, a significant reduction (P < 0.05) in the number of infected chickens and in the number of SE in the cecal contents was observed at 10 wk of age in Trial 2. The enrofloxacin treatment did not increase opportunistic colonization by SE due to the use of the antibiotic in either trial. The plasma and intestinal immunological responses were not significant at the early age (up to 12 wk) of the birds. The use of enrofloxacin, followed by NAGF, could aid the elimination of SE from young chicks persistently infected at an early age. The combined treatment, compared with enrofloxacin alone, protected chickens from reinfection by 40%. PMID- 11055846 TI - Effect of Eimeria acervulina infections on plasma L-arginine. AB - As part of a program to study the pathological effects of coccidia infections on growth, we have examined the relationship of plasma L-arginine (ARG) levels to infective doses of Eimeria acervulina and infection-associated changes in weight gain, plasma carotenoids, and plasma NO2- + NO3-. Chickens consuming a starter ration containing 1.68% ARG were infected with a range of doses of E. acervulina. At 6 d postinoculation (PI), weight gains were significantly reduced by infections with 5 x 10(5) and 1 x 10(6) oocysts per chick (OPC). Gross lesion scores of chickens infected with 5 x 10(4) through 1 x 10(6) OPC were significantly greater than scores of chicks infected with 1 x 10(3) OPC. Compared with levels from uninfected controls, plasma NO2- + NO3- concentrations were significantly increased by infection with 5 x 10(5) and 1 x 10(6) OPC, plasma concentrations of ARG were significantly decreased by infection with 5 x 10(4) through 1 x 10(6) OPC, and plasma carotenoids were significantly decreased by all infection doses. Plasma arginine was significantly correlated with plasma carotenoids (P > 0.0187), but not with infection dose or weight gain; plasma NO2- + NO3- was positively correlated (P > 0.0043) with infection dose and negatively correlated (P > 0.0158) with weight gain. Regression analysis of the measured variables indicated that the strongest relationship existed between plasma ARG and carotenoids. This finding suggests that in this infection model, reduction in plasma ARG is most likely associated with nutrient malabsorption that accompanies infection and is likely not significantly impacted by synthesis of nitric oxide that is associated with the immune response. PMID- 11055847 TI - The dynamics of antibody response to Escherichia coli vaccination in meat-type chicks. AB - The dynamics of serum antibody (Ab) response in young broilers were studied in lines divergently selected for high (HC) or low (LC) Ab response to Escherichia coli vaccination at an early age, and their cross (HL). Chicks were divided into three vaccination-age (VA) groups: 8, 10, and 12 d of age (VA8, VA10, and VA12, respectively). Antibody response was determined five times for each chick, at 6, 8, 10, 12, and 14 d postvaccination (dPV). The effects of line, VA, and dPV on Ab titers were highly significant. The HC and LC chicks exhibited the highest and lowest mean titers, respectively, in all VA groups. The HL chicks exhibited midparent Ab values for all VA and dPV combinations, indicating additive inheritance of early Ab production. In LC, the highest mean Ab titer was obtained on Day 26 (14 dPV of the VA12 group), whereas in HC, the same titer had already been obtained on Day 18 (VA8-10 dPV and VA10-8 dPV combinations). The VA8 and VA12 chicks differed markedly in their Ab titer dynamics curves, and the VA10 chicks exhibited an intermediate curve. The three VA groups exhibited a similar change in Ab level from 6 to 10 dPV, but they differed in Ab change from 10 to 14 dPV. This significant dPV x VA interaction suggests that the VA12 and VA10, but not VA8, chicks maintained the capability to produce persisting Ab. PMID- 11055848 TI - Reduction of mortality in specific-pathogen-free layer chickens by a caprine serum fraction after infection with Pasteurella multocida. AB - Caprine serum was fractionated by size, and its proteinaceous material <8,000 Da [caprine serum fraction immunomodulator 2 (CSF-I2)] was evaluated for its ability to impart immunoresistance to specific-pathogen-free (SPF) layer chickens. The SPF layers were challenged with 18 to 30 cfu of Pasteurella multocida X-73 (serotype 1) at 5 wk of age. A high degree of mortality was apparent 24 and 48 h later (62+/-14% and 88+/-7%, respectively). Mortality observed after 48 h was minimal. Noting the rapid onset of mortality, we administered CSF-I2 (material that expressed no direct antimicrobial activity but was believed to be an immunostimulant) 1 d before challenge and coincident to time of challenge. The group of birds that received CSF-I2 (either 5 or 10 mg per administration) expressed significant reduction in mortality throughout the 1-wk study period. Reduction in mortality appeared to be dose dependent. Birds that received two administrations of 10 mg CSF-I2 had significantly fewer deaths than did the group of birds that received half that amount. No deaths were recorded through 24 h, whereas, at 48 h, the percentage mortality was 13 in CSF-I2-treated birds. This study demonstrates that one or more small molecular weight compounds isolated from caprine serum were able to reduce mortality in SPF layers infected with Pasteurella multocida. PMID- 11055849 TI - A modified method of shell windowing for producing somatic or germline chimeras in fertilized chicken eggs. AB - Stage X chick blastoderms following oviposition were accessed via a small window in the egg. Windowing, however, substantially reduces the hatchability of eggs containing early embryos. For example, only 32 of 389 (8.2%) eggs hatched after standard windowing with or without irradiation or injection. Ex ovo culture systems can overcome this problem but are labor intensive. A modification of a standard windowing technique has yielded an average hatch rate of 32% for 892 windowed eggs independent of incubator type, gamma-irradiation, or injection of the embryo. This was a fourfold increase over a standard windowing method. Similar hatch rates were observed using fertile eggs from five chicken lines [Barred Plymouth Rock (BR), Athens-Canadian (AC), Line 0, SPAFAS, and commercial White Leghorns (WL)]. The modification involves covering the egg shell membrane with PBS after grinding away the shell and before piercing the membrane. The window is then sealed by overlaying with fresh shell membrane and cementing it in place once it has dried. The method has been used successfully for the production of somatic and germline chimeras because donor BR blastodermal cells injected into Stage X, gamma-irradiated recipient embryos from WL or AC yielded a hatch of 33.7%, of which 42.3% were feather chimeras. Two of 11 cockerels tested were germline mosaics bearing at least 1% BR sperm. The modified windowing technique may be broadly applicable in emerging technologies in avian transgenesis and development. PMID- 11055850 TI - Comparison of the efficacies of a novel aspergillus niger mycelium with separate and combined effectiveness of phytase, acid phosphatase, and pectinase in dephosphorylation of wheat-based feeds fed to growing broilers. AB - Efficacies of phytase, phosphorolytic enzymes (phytase + acid phosphatase), an enzymic "cocktail" (phytase + acid phosphatase + pectinase + citric acid), a novel Aspergillus niger (fungal) mycelium (FM), and FM enriched in phytase and antioxidants were investigated in growing broilers (Days 1 to 21) fed wheat-based diets. Broilers were fed the following seven diets at 0.69% Ca: 1) a negative control diet, 0.17% nonphytate P (NPP); 2) Diet 1 + 750 phytase units/kg diet; 3) Diet 1 + 750 phytase units + 3,156 units acid phosphatase/kg diet; 4) Diet 1 + 750 phytase units + 3,156 acid phosphatase units + 1,900 units of pectinase/g diet + 3% citric acid; 5) Diet 1 + 4% FM; 6) Diet 1 + 4% FM + 1,300 phytase units + 2% ascorbic acid and 1% of glucose oxidase; and 7) a positive control diet (Diet 1 + 0.24% NPP from dicalcium phosphate). The dietary treatments were fed to four pen replicates of eight birds each. Prior to feed formulation, mycelium and antioxidants dosages were optimized on Diet 1 by an in vitro technique and an experimental design module of a statistical software package. Phytase addition increased BW gain (BWG), feed intake, and P retention. Subsequent addition of acid phosphatase resulted in further increases in BWG, feed intake, and toe ash and reduced digesta viscosity; however, neither P nor Ca retention were improved. Body weight gain and feed intakes superior to those found in chicks fed Diet 7 were observed in birds receiving the cocktail of enzymes (Diet 4) or FM. Chicken fed Diet 6 had the highest percentage of toe ash and retained 76 and 51% of P and Ca, respectively. Supplementation of wheat-based 0.17% NPP diets with FM increased bursa of Fabricius weights and reduced the intestinal surface covered by Peyer's patches. PMID- 11055851 TI - Effects of low phytic acid corn on phosphorus utilization, performance, and bone mineralization in broiler chicks. AB - In vivo and in vitro experiments were conducted to determine whether P in a low phytate corn (LPC) containing the lpa 1-1 allele is more available than P in a near-isogenic wild-type corn hybrid (NC). The LPC was analyzed to contain 0.18% nonphytate P and 0.26% total P (TP), whereas NC contained 0.05% nonphytate P and 0.25% TP. For these studies, nonphytate P was considered to be available P (AP). In the in vivo study, 150 1-d-old male chicks were randomly assigned to five treatments (six pens of five chicks each) for 21 d. The dietary treatments included: A) a diet containing 60% NC, 0.2% AP, and 0.8% Ca; B) a diet containing 60% LPC, 0.28% AP, and 0.8% Ca; C) an NC diet similar to Diet A, but with KH2PO4 added to increase the AP to 0.28% to match the AP in Diet B; D) an LPC diet containing 0.45% AP and 1% Ca; and E) an NC diet supplemented with KH2PO4 to provide 0.45% AP and 1% Ca. Diets A, B, and C were semipurified diets, with corn being the sole source of phytate. The only differences between Diets A and B were the source of corn and the amount of AP present in the diets. The levels of AP in these diets were deficient in order to measure the animal response to the different levels of AP. Diets D and E were typical corn-soybean meal diets, and were formulated to contain an optimal level of AP. Performance and bone ash were similar (P > 0.05) in chicks fed Diets B and C and in chicks fed Diets D and E. Chicks fed LPC diets (B and D) retained more P (P < 0.05) than chicks fed NC diets (C and E). Chicks fed Diet B had significantly higher (P < 0.05) Ca retention compared with chicks fed Diet A. An in vitro digestion procedure that simulated the physiological conditions of the gastrointestinal tract of broilers was used to determine P release from LPC and NC. Results showed that 65% (1,420 mg/kg) of the TP in LPC was released, compared with 23% (543 mg/kg) from NC. Results of these experiments indicate that the P in LPC is more available than the P in NC, and reducing the phytate content did not compromise the nutritional value of LPC. The increased P retention in chicks fed LPC suggests that substituting LPC for NC leads to a reduction in manure P. Also, the in vitro procedure accurately predicted differences in in vivo P availability between the two corns. PMID- 11055852 TI - Nonphytate phosphorus requirement and phosphorus excretion of broiler chicks fed diets composed of normal or high available phosphate corn with and without microbial phytase. AB - A study was conducted to evaluate the ability of the young (0 to 3 wk) broiler chicken to utilize the P provided by a high available P corn [HAPC; 0.27% total P and 0.17% nonphytate P] in comparison with yellow dent corn (YDC; 0.23% total P and 0.03% nonphytate P), and to determine the extent to which supplementation with exogenous phytase enzyme could reduce the demands for dietary P and subsequently reduce P excretion. Diets prepared using the two types of corn differed in the amount of phytate-bound P, with the HAPC diets containing approximately 50% less phytate-bound P. Treatment diets were prepared by varying the amount of dicalcium phosphate, and ranged from 0.10 to 0.50% nonphytate P for YDC diets, and from 0.18 to 0.50% nonphytate P for HAPC diets. Sublots of each diet were supplemented with 800 units/kg phytase. Each diet was fed to six pens of five male chicks of a commercial broiler strain from 1 to 21 d of age. Regression analysis was used to estimate nonphytate P requirements for each corn type with and without phytase supplementation. The greatest need for nonphytate P was for maximum tibia ash, with requirements of 0.39, 0.29, 0.37, and 0.32% in diets with YDC, YDC plus phytase, HAPC, and HAPC plus phytase, respectively. Addition of phytase liberated approximately 50% of the phytate-bound P from each diet. These levels were sufficient to support body weight, feed conversion, and livability. Fecal P content of broilers fed diets with YDC at the NRC (1994) recommended level of 0.45% nonphytate P was 1.21%, whereas at the respective requirement points indicated above, the P content was 1.09, 0.87, 0.78, and 0.64% in feces from broilers fed diets with YDC, YDC plus phytase, HAPC, and HAPC plus phytase, respectively. Thus, fecal P output could be reduced while maintaining optimum performance by the use of reduced dietary nonphytate P, introduction of HAPC, and phytase supplementation. One of the greatest benefits of phytase supplementation appeared to be maintaining livability at lower dietary levels of nonphytate P. PMID- 11055853 TI - The developmental expression of acyl-coenzyme A:cholesterol acyltransferase in the yolk sac membrane, liver, and intestine of developing embryos and posthatch turkeys. AB - Acyl-coenzyme A:cholesterol acyltransferase (ACAT) catalyzes the formation of cholesterol esters (CE) from free cholesterol and fatty acyl-coenzyme A. This experiment was conducted to study the ontogeny of ACAT activity in the yolk sac membrane, liver, and intestine during embryonic development and early posthatch growth of turkeys. The ACAT activity was measured on tissue samples collected at 3-d intervals from embryonic Day (ED 13) 13 through 6 d posthatch (PD 6). The ACAT activity (pmol/mg microsomal protein per min) in the yolk sac membrane increased form 840 pmol at ED 13 to 2,497 pmol at ED 22, and subsequently declined to a very low level by PD 3. The high level of enzyme activity at ED 22 is concomitant with the large quantity of CE formed within the yolk sac membrane at this developmental age. Liver ACAT activity increased from 60 pmol at ED 13 to 242 to 243 pmol at ED 25 and PD 3, followed by a decline to 130 pmol by PD 6, mirroring the peak in hepatic CE concentration. This suggests that even during incubation, the liver plays a significant role in lipid metabolism. Intestinal ACAT specific activity increased from 14 pmol (ED 16) to 44 pmol (ED 25), and then declined to 23 pmol by hatch (ED 28), with no further decline through PD 6. Total intestinal ACAT activity (pmol per intestine/min) increased, however, from ED 16 through PD 6. This increase in activity suggests that the total capacity for cholesterol esterification increases during the course of incubation and shortly after hatching. PMID- 11055854 TI - Muscle protein turnover during early development in chickens divergently selected for growth rate. AB - To explore the mechanisms involved in the genetic control of muscle growth and protein gain, protein metabolism was assessed in the pectoralis major muscle of two chicken lines selected for either fast or slow growth. Protein synthesis was measured in vivo at various ages from 1 to 4 wk, using a flooding dose of L-[4 3H] phenylalanine. Protein degradation was estimated as the difference between synthesis and deposition. Over the experimental period, BW were about 2-fold greater (P < 0.001), and pectoralis major muscle weights were 2.4- to 3.6-fold higher (P < 0.001), in chicks from the fast-growing line (FGL) than those from the slow-growing line (SGL). Independent of age, absolute rates of protein deposition, synthesis, and breakdown were higher in FGL than in SGL chickens. Fractional rates of muscle protein synthesis clearly decreased with age. When comparing birds of the same age, fractional rates of muscle protein synthesis tended to be lower in the FGL. Fractional degradation rates (KD) were significantly lower in FGL chickens during the first 2 wk of post-natal growth, whereas KD were similar between lines in older chickens. In this experimental model of chicken lines divergently selected for BW, the greatest line-related difference in muscle protein metabolism was in KD, and was observed in the early growth phases. PMID- 11055855 TI - Interaction between ambient temperature and supplementation of synthetic amino acids on performance and carcass parameters in commercial male turkeys. AB - An experiment with male turkeys was conducted to test the hypothesis that turkey production performance responds positively to extra crystalline amino acid supplementation (lysine, methionine, and threonine) when subjected to a high ambient temperature regimen (HT) in the grower period. Two diets were formulated to provide lysine, methionine, and threonine concentrations that either 1) met the breeder recommendations or 2) contained 10% higher lysine and methionine concentrations from 22 to 134 d of age and 10% higher threonine concentration from 22 to 68 d of age. Both diets were fed at two temperatures (15 or 25 C) from 42 d of age onward. At 134 d of age, turkeys on the HT had generally lower BW than those on the low temperature regimen (LT). Up to 68 d of age and from 106 to 134 d of age, feed intake of turkeys on the HT was significantly lower than that of turkeys on the LT. Up to 42 d of age, feed conversion ratio (FCR) of turkeys on the HT were significantly lower than those of turkeys on the LT. Significant treatment interactions were observed from 22 to 41 d of age. Turkeys fed the amino acid-supplemented diets on the LT had significantly reduced FCR, whereas those on the HT did not respond. From 69 to 105 d of age, turkeys on the HT that were fed the supplemented diets had significantly increased FCR, but there were no dietary effects among turkeys on the LT. There were no consistent diet effects on growth performance or carcass yields. Breast meat yields of turkeys on the LT were higher (33.5 vs 32.1%), and drum yields were lower (12.7 vs 13.0%), than those of turkeys on the HT. There were no significant amino acid balance x ambient temperature effects on processing yields. The hypothesis of this experiment could be rejected as production performance did not respond positively to extra supplementation of lysine, methionine, and threonine when subjected to an HT. PMID- 11055856 TI - Influence of dietary protein level on the broiler chicken's response to methionine and betaine supplements. AB - Two experiments were conducted to compare broiler chicken responses to methionine and betaine supplements when fed diets with low protein and relatively high metabolizable energy levels (17%, 3.3 kcal/g) or moderate protein and lower metabolizable energy levels (24%, 3.0 kcal/g), resulting in different levels of carcass fat. In Experiment 1, the basal diets were formulated with corn, soybean meal, poultry by-product meal, and poultry oil. In Experiment 2, glucose monohydrate was also added, so that identical amino acid profiles could be maintained in the 17 and 24% protein diets. On average, feeding the 17 vs. 24% protein diet decreased 21-d body weight gain by 20%, increased feed conversion ratio (FCR) by 13%, and increased abdominal fat pad weight by 104%. Methionine and betaine supplements improved the performance of chicks fed the 24% protein diet in both experiments, as indicated by body weight gain and FCR. Only supplementary methionine increased performance of chicks fed 17% protein diets, and then only in Experiment 2. Neither methionine nor betaine decreased abdominal fat pad size in either experiment. Methionine supplementation decreased relative liver size and increased breast muscle protein. Both methionine and betaine increased sample feather weight, but when expressed as a percentage of body weight, no significant differences were detected. It is concluded that increasing carcass fat by manipulating percentage dietary protein level or amino acid balance does not influence betaine's activity as a lipotropic agent. PMID- 11055857 TI - The effect of varying mix uniformity (simulated) of phytase on growth performance, mineral retention, and bone mineralization in chicks. AB - An 18-d experiment was conducted to determine the effect of varying mix uniformity of phytase on growth performance, mineral retention, and bone mineralization in chicks. Chicks (initial and final weights were 74.5 and 803.3 g) were allotted to seven treatments with six (Treatment 1) or seven (Treatments 2 to 7) replicates of seven chicks per replicate in a completely randomized design. Varying mix uniformity of phytase was simulated by alternately providing two diets with two different concentrations of microbial phytase; the diets were switched every 24 h. Treatments were: 1) positive control (CON) (Ca, 1.0%; available phosphorus (aP), 0.45%), 2) negative control (NEG) (Ca, 0.9%; aP, 0.35%), 3) NEG + 600 phytase units (FTU) daily (CV0), 4) NEG + 500 or 700 FTU (CV17), 5) NEG + 400 or 800 FTU (CV34), 6) NEG + 200 or 1,000 FTU (CV69), or 7) NEG + 0 or 1,200 FTU (CV103). Gain, feed intake, and bone breaking strength were similar (P > 0.15) in the CON and CV0 treatments, but these response variables were decreased in the NEG treatment (P < 0.01). Gain:feed was not affected by treatment (P = 0.15). Bone ash was decreased (P < 0.02) by the NEG and CV0 treatments compared with the CON diet, but chicks fed the CV0 diet had greater bone ash than those fed the NEG (P < 0.01) diet. Increasing FTU CV decreased bone breaking strength and bone ash (P < 0.01). Calcium and phosphorus retention (P < 0.08) and gain (P < 0.09) were numerically decreased, and phosphorus excretion was numerically increased (P < 0.07) as FTU CV increased. The difference between the CV0 and CV103 treatments was significant only for bone breaking strength and ash (P < 0.01). In conclusion, increasing phytase CV had little effect on growth performance, whereas bone ash and breaking strength and calcium and phosphorus retention and excretion decreased only at the most extreme CV. PMID- 11055858 TI - Development of the reproductive system in turkeys with a high or low susceptibility to prolapse of the oviduct. AB - Lines of turkeys selected for rapid growth and high meat yield have an increased incidence of prolapse of the oviduct compared with unselected or traditional strains of turkeys. The development of the reproductive system and changes in plasma estrogen concentrations were compared in sire line and traditional turkeys with the aim of identifying any morphological or hormonal differences that could be associated with the high incidence of prolapse in the male line. Four turkeys from each strain were killed weekly from 0 to 7 wk postphotostimulation, and samples from prolapsed birds were obtained from field cases. There were no differences in the rate of development of the ovary, oviduct, uterus, vagina, sphincter ani muscle, or muscular cord of the ventral ligament between the two strains that could predispose the sire line to prolapse. Histological investigation of the uterus, vagina, muscular cord of the ventral ligament, and sphincter ani muscle 5 wk postphotostimulation in traditional, sire line, and prolapsed sire line turkeys did not reveal any differences that could be associated with prolapse. No prelay peak in plasma estradiol concentration was observed in either strain, and there was no evidence to suggest that plasma estradiol was higher in the sire line compared with the traditional turkeys. It was concluded that prolapse of the oviduct in sire line turkeys was not associated with any anatomical abnormalities or high plasma estradiol during reproductive development. PMID- 11055859 TI - Effect of low extraction temperatures on microbiological quality of rendered chicken fat recovered from skin. AB - Ground or finely homogenized skin, inoculated with circa 7 log10 cfu/g of an Acinetobacter sp., Brochothrix thermosphacta, Candida tropicalis, Debaryomyces hansenii, Enterobacter agglomerans, Enterococcus faecalis, a Lactobacillus sp., or Pseudomonas fluorescens, or not inoculated, was heated to 50 or 80 C to release fat from adipocytes, and the released fat was separated by centrifugation. Extraction at 80 C resulted in nearly complete inactivation of indigenous and inoculated flora, resulting in microbiological counts generally below detection level in skin residue and rendered fat. In contrast, large numbers of organisms (3.69 to 7.28 log10 cfu/g) survived the 50 C extraction process. Even though the majority (91.5 to 99.9%) of these organisms remained in the residual skin at the time of fat separation, some organisms were also found occasionally in fat at concentrations of 2.85 to 3.74 log10 cfu/g. Therefore, an additional step such as flash pasteurization is recommended for safety, should extraction temperatures below 80 C be selected. PMID- 11055860 TI - Detection of chelonid herpesvirus DNA by nonradioactive in situ hybridization in tissues from tortoises suffering from stomatitis-rhinitis complex in Europe and North America. AB - Chelonid herpesvirus (ChHV) infection in tortoises associated with stomatitis rhinitis complex is a severe, mostly epizootic disease characterized by proliferative and diphtheroid-necrotizing glossitis, pharyngitis, rhinitis, and tracheitis, often occurring with pneumonia and encephalitis. The UL5 gene from a German ChHV isolate was used to generate a digoxigenin-labeled 307-base-pair DNA probe by polymerase chain reaction (PCR). ChHV DNA was detected in paraffin embedded tissues of five naturally infected tortoises (two Afghan tortoises [Testudo horsfieldii], USA; two Hermann's tortoises [Testudo hermanni], Switzerland; one T. hermanni, Germany) by means of in situ hybridization (ISH) and PCR. Distribution of ChHV DNA exhibits many characteristics of alphaherpesvirus but also some characteristics of betaherpesvirus infections. The amino acid sequence of a portion of the ChHV UL5 homolog exhibited more than 50% similarity to alphaherpesvirus UL5 proteins. Nuclear hybridization signals were detected in epithelial cells of the lingual mucosa and glands. Furthermore, ChHV DNA was observed in tracheal epithelium, pneumocytes, hepatocytes, the renal tubular epithelium, cerebral glia cells and neurons, and intramural intestinal ganglia. ChHV DNA in endothelial cells of many organs underlines the systemic character of the disease. Importantly, ChHV DNA was detected by ISH in multiple tissues of tortoises originating from different geographic provenances. This indicates a high degree of conservation of the UL5 gene fragment among viruses prevalent in tortoises on different continents. With the described ISH, a molecular biological tool is available for rapid and specific diagnosis of ChHV infections and, more importantly, comparative pathogenetic studies of ChHV isolates from geographically unrelated regions. PMID- 11055861 TI - Relationship of lymphoid lesions to disease course in mucosal feline immunodeficiency virus type C infection. AB - Feline immunodeficiency virus (FIV) infection typically has a prolonged and variable disease course in cats, which can limit its usefulness as a model for human immunodeficiency virus infection. A clade C FIV isolate (FIV-C) has been associated with high viral burdens and rapidly progressive disease in cats. FIV-C was transmissible via oral-nasal, vaginal, or rectal mucosal exposure, and infection resulted in one of three disease courses: rapid, conventional/slow, or regressive. The severity of the pathologic changes paralleled the disease course. Thymic depletion was an early lesion and was correlated with detection of FIV RNA in thymocytes by in situ hybridization. The major changes in thymic cell populations were depletion of p55+/S100+ dendritic cells, CD3- cells, CD4+/CD8- cells, and CD4+/CD8+ cells and increases in apoptosis, CD45R+ B cells, and lymphoid follicles. In contrast to thymic depletion, peripheral lymphoid tissues often were hyperplastic. Mucosally transmitted FIV-C is thymotropic and induces a spectrum of lymphoid lesions and disease mirroring that seen with the human and simian immunodeficiency virus infections. PMID- 11055862 TI - Comparative immunohistopathology in pigs infected with highly virulent or less virulent strains of hog cholera virus. AB - Eight pigs were inoculated subcutaneously with a highly virulent hog cholera virus (HCV) strain ALD. The infected pigs developed severe illness and became moribund on postinoculation day (PID) 7 or PID 10. Histologic lesions were characterized by severe generalized vasculitis, necrosis of lymphocytes, and encephalitis. HCV antigen was detected in crypt tonsilar epithelial cells, macrophages, and reticular endothelial cells of lymphoid tissues. Antigen localization corresponded well with histologic lesions. Five pigs were inoculated with less virulent HCV Kanagawa/74 strain and were euthanatized on PID 30. All five infected pigs recovered from the illness but became stunted. They also had a slight follicular depletion of lymphocytes, histiocytic hyperplasia, and hematopoiesis in the spleen. Less virulent HCV antigen was observed in the tonsils, kidneys, pancreas, adrenal glands, and lungs. Although antigen localization was less associated with histologic lesions, immunoreactivity was stronger than that in the pigs infected with the ALD strain of HCV. An almost complete loss of B lymphocytes was recognized in pigs infected with the ALD strain and was correlated with follicular necrosis in lymphoid tissues. Loss of B lymphocytes was not prominent in the pigs infected with Kanagawa/74 strain. The number of CD4+ and CD8+ T lymphocytes was significantly higher than that in the noninfected control pigs. PMID- 11055863 TI - Glomerular lesions in renal biopsies of Saimiri boliviensis (primate) examined by light and electron microscopy and immunohistochemistry. AB - In order to determine the existence of glomerular lesions in Saimiri boliviensis, renal biopsies were performed in 20 clinically healthy animals of similar age and both sexes. Biopsies were obtained by laparotomy with a Tru-Cut biopsy needle. Mesangioproliferative glomerulonephritis characterized by an increased number of mesangial cells and increased mesangial matrix was present in 35% of the animals. Proliferative glomerulonephritis characterized by increased numbers of epithelial and endothelial cells with narrowed capillary lumen, and membranous glomerulonephritis characterized by diffuse thickening, wrinkling, and occasional lamellation of basement membranes, were present in 15% of the samples. Ultrastructural features included increased mesangial matrix, fusion of the visceral epithelial foot processes, thickened glomerular basement membranes, and incipient lamellation. Immunohistochemical examination revealed granular deposits of immunoglobulin M in the cytoplasm of mesangial cells and in the mesangial matrix in 50% of the samples. PMID- 11055864 TI - Reduced formation of granulomata in gamma(delta) T cell knockout BALB/c mice inoculated with Mycobacterium avium subsp. paratuberculosis. AB - The role of gamma(delta) T cells in the bovine immune response to Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) infection is poorly understood. Accordingly, using BALB/c mice that are innately susceptible to M. paratuberculosis, we compared wild-type and gamma(delta) T cell knockout BALB/c mice to study the protective roles of gamma(delta) T cells in M. paratuberculosis infection. Ten-week-old mice were inoculated intraperitoneally with either a low dose (4 x 10(6) colony-forming units [CFU]/mouse) or a high dose (4 x 10(9) CFU/mouse) of M. paratuberculosis strain ATCC 19698. Histopathologic and morphometric examinations showed reductions in the number and area of granulomatous lesions in the liver of the knockout mice at 18 weeks after inoculation with either the low or the high dose of the mycobacteria. Furthermore, at 18 weeks after inoculation, the bacterial load in the spleens of the knockout mice inoculated with the high dose was significantly lower than that of wild-type mice. No differences were found in bacterial load between the knockout and the wild-type mice in the low-dose groups. In contrast, in the livers of wild-type mice inoculated with either the low or high mycobacterial dose, increased areas of epithelioid granulomata were observed and the granulomata became disseminated widely during the experimental period. These findings in model mice suggest that gamma(delta) T cells, rather than restricting mycobacterial growth, may play a crucial role in development of epithelioid granulomata similar to those seen consistently in bovine paratuberculosis. The results of this study may have relevance to our understanding of the pathogenesis of paratuberculosis in ruminants, in which a prominent number of gamma(delta) T cells exist in the lymphoid system. PMID- 11055865 TI - Persistent hyperplastic tunica vasculosa lentis and persistent hyperplastic primary vitreous in transgenic line TgN3261Rpw. AB - Persistent hyperplastic tunica vasculosa lentis and persistent hyperplastic primary vitreous are congenital ocular anomalies that can lead to cataract formation. A line of insertional mutant mice, TgN3261Rpw, generated at the Oak Ridge National Laboratory in a large-scale insertional mutagenesis program was found to have a low incidence (8/243; 3.29%) of multiple developmental ocular abnormalities. The ocular abnormalities include persistent hyperplastic primary vitreous, persistent hyperplastic tunica vasculosa lentis, failure of cleavage of the anterior segment, retrolental fibrovascular membrane, posterior polar cataract, and detached retina. This transgenic mouse line provides an ontogenetic model because of the high degree of similarity of this entity in humans, dogs, and mice. PMID- 11055866 TI - The pathology of spontaneous paratuberculosis in the North American bison (Bison bison). AB - Gross and histopathologic examinations were performed on 70 North American bison (Bison bison) from a Mycobacterium avium paratuberculosis culture-positive herd. The bison examined were part of a breeding herd totaling 2,800 animals. Eight of 70 (11%) animals had gross findings of intestinal mucosal thickening, and 16 of 70 (23%) of the animals had enlarged mesenteric lymph nodes. Histologic lesions compatible with Johne's disease were diagnosed in 30 of 70 (43%) bison on the basis of the demonstration of noncaseating granulomatous inflammatory infiltrates and of one or more acid-fast bacilli characteristic of Mycobacterium avium paratuberculosis. A suspicious diagnosis of Johne's disease was obtained in 11 of 70 (16%) bison on the basis of the observation of noncaseating granulomatous inflammatory infiltrates without demonstrable acid-fast bacteria. Twenty-nine of 70 (41%) animals were assessed as histologically paratuberculosis free. Histologic results were compared to Johne's disease tests such as culture, serology, and polymerase chain reaction, which were performed on some of the cohort animals. PMID- 11055867 TI - Granular cell lesions in the distal female reproductive tract of aged Sprague Dawley rats. AB - During the review of a rat carcinogenicity study, a spectrum of granular cell lesions was recognized in the distal female reproductive tract. To verify the diagnoses, cell populations of nine granular cell alterations of the cervix or vagina were characterized immunohistochemically and four were evaluated ultrastructurally. Immunoreactivity was demonstrated in 8/9 cases with S100 protein, 6/9 cases with neuron-specific enolase, and 7/9 cases with Leu-7. Granular cells were negative for smooth muscle-specific actin and calretinin. The immunohistochemical profile of these lesions was similar to that previously reported in other species, including humans. Ultrastructurally, as expected many lysosomal bodies were present in the cytoplasm of granular cells in all specimens evaluated. Based on the detailed evaluation of a series of lesions, we adopted the following diagnostic criteria and nomenclature for the granular cell changes of the female reproductive tract of rats. Granular cell aggregates were non-space occupying lesions composed of clusters of typical granular cells. Benign granular cell tumors were space occupying lesions that typically contained prominent interstitial collagen and were either discrete masses or were difficult to discern from the surrounding tissues. Some benign tumors also contained foci of spindle cells with decreased granularity. Malignant tumors exhibited pleomorphism and an increased nucleus: cytoplasm ratio morphologically but had the same biologic behavior as the benign tumors. We applied these diagnostic criteria during the review of controls from 9 carcinogenicity studies. Up to 23% of control females in those carcinogenicity studies had granular cell lesions that could be classified into one of the three categories. Granular cell lesions appear to be common in the cervix/vagina of the Sprague-Dawley rat, and tumors may develop from granular cell aggregates. PMID- 11055868 TI - Isolation and characterization of an antigenically distinct 68-kd protein from nonviral intracytoplasmic inclusions in Boa constrictors chronically infected with the inclusion body disease virus (IBDV: Retroviridae). AB - The relationship between a retroviral infection and the development of nonviral intracytoplasmic inclusion bodies was studied in a Boa constrictor model. Twelve juvenile age- and size-matched inclusion body disease (IBD)-negative boas were randomly divided into three groups. Each group was inoculated intraperitoneally with 1 ml of an IBD virus (IBDV)-infected liver homogenate or 1 ml of normal boa liver homogenate (sham-inoculated control) or was left untreated. All boas were monitored for development of IBD by daily examination and serial liver biopsy over 1 year. The 4 IBDV-inoculated boas became IBDV and inclusion positive by 10 weeks postinoculation. The average size and density of inclusion bodies increased with the duration of infection. Ultrastructurally, inclusion bodies <2 microm in diameter consisted of intracytoplasmic aggregates of granular electron-dense material that were not membrane limited. Larger inclusions (3-6 microm in diameter) were characterized as membrane-bound aggregates of amorphous to granular electron-dense material admixed with membranelike fragments. The sham inoculated and untreated control snakes did not become inclusion or IBDV positive. Direct comparison of the protein electrophoretograms of IBDV-infected and normal boa tissues demonstrated a prominent 68-kd protein band unique to infected inclusion-positive tissues. Monoclonal antibodies directed against the 68-kd protein band specifically labeled inclusion bodies. The results of this study demonstrate that IBD inclusions represent an intracytoplasmic accumulation of an antigenically distinct IBDV-associated protein. PMID- 11055869 TI - Endocardial ossifying myxoma of the right atrium in a cat. AB - Primary cardiac neoplasms are rare in humans and animals. In humans, the most common primary cardiac tumor is the myxoma, which is frequently found in the left atrium. Cardiac myxoma has been reported in the dog but not in the cat. We describe the gross, immunohistochemical, and light microscopic examination of a myxoma in the right atrium of a 6-year-old domestic shorthair cat. Histologically, the tumor consisted predominantly of mesenchymal cells with several foci of bone and cartilage present. The tumor was encapsulated and benign. PMID- 11055870 TI - An encephalopathy with argyrophilic inclusions in a Holstein-Friesian cow. AB - The brain of a 6-year-old Holstein cow, which showed progressive neurologic symptoms during several months, was examined by histopathologic methods. Many round or oval-shaped cytoplasmic inclusions were observed, mainly in neurons of the temporal lobe and the hippocampus. Those inclusions were faintly eosinophilic with hematoxylin and eosin and positive with Bielschowsky's silver stain. Immunohistochemically, the inclusions were recognized by antiubiquitin and antiphosphorylated tau antibodies. Ultrastructurally, the inclusions were globular and well demarcated from the rest of the cytoplasm, lacked limiting membranes, and were mainly composed of straight fibrils about 15 nm in width. The structure of the inclusions was similar to that of Pick bodies in Pick's disease of humans. The pathogenesis of this bovine condition is not known. PMID- 11055871 TI - Progression of an orbital T-cell rich B-cell lymphoma to a B-cell lymphoma in a dog. AB - An 11-year-old Shetland Sheepdog was presented for exophthalmos caused by a locally extensive, poorly defined mass located behind the right eye. The primary orbital mass was identified by light microscopy and immunohistochemistry as a T cell rich B-cell lymphoma (TCRBCL) composed predominantly of BLA.36-positive large neoplastic lymphoid cells admixed with fewer CD3- and CD79a-positive small lymphocytes. The dog was treated for lymphoma, but 6 months after presentation it was euthanatized for suspected hepatic and gastrointestinal metastasis. Gross findings revealed an enlarged liver with multiple well-demarcated, randomly distributed 0.1-1.5-cm white nodules, five firm white submucosal jejunal nodules, and ileocecal, mediastinal, and hilar lymphadenopathy. Metastatic liver lesions consisted of sheets of monomorphic large neoplastic lymphoid cells that effaced and expanded portal and centrilobular zones. These cells were morphologically similar to the large neoplastic cells of the original orbital tumor and were CD3 negative and variably BLA.36-positive, consistent with B-cell lineage. Similar cells comprised the jejunal nodules and effaced the lymph nodes. The progression of TCRBCL to a diffuse B-cell lymphoma in this case is consistent with reported human cases and has not been previously reported in the dog. PMID- 11055872 TI - Leukoencephalitis and vasculitis with perivascular demyelination in a Weimaraner dog. AB - Aseptic and noninfectious diseases of the central nervous system are being recognized with increasing frequency. After multiple episodes of neurologic illness, this 7-year-old Weimaraner dog was euthanatized and submitted for postmortem examination. Lesions in the central nervous system were found mainly in the white matter of the cerebral cortex and cervical spinal cord and represented acute and more chronic injury. Necrotizing vasculitis with fibrinoid change and a marked neutrophilic infiltrate dominated the acute lesions. More chronic changes consisted of perivascular demyelination and accumulation of foamy macrophages with positive staining for myelin. An immune-complex (Arthus-type) vasculitis is suspected. PMID- 11055873 TI - Pulmonary cryptosporidiosis in simian immunodeficiency virus-infected rhesus macaques. AB - Cryptosporidiosis is a common opportunistic infection in the gastrointestinal tract of human and nonhuman primates with AIDS. Pulmonary infection associated with Cryptosporidium spp. has not been previously reported in monkeys. Two macaques experimentally infected with simian immunodeficiency virus (SIV) had lesions containing cryptosporidial organisms involving the trachea, lungs, bile ducts, pancreas, and intestine. The pulmonary sections revealed moderate to severe bronchopneumonia associated with cryptosporidiosis. Numerous 2-4 microm oval Cryptosporidium spp. organisms were present in the cytoplasm of alveolar macrophages, multinucleated giant cells, and intestinal epithelial cells. Giant cells were positive for SIV by in situ hybridization. These are the first reported cases of cryptosporidiosis with involvement of pulmonary parenchyma in SIV-infected macaques. PMID- 11055874 TI - Brain lesions and transmission of experimental equine herpesvirus type 9 in pigs. AB - We demonstrated that pigs are susceptible to acute infection by equine herpesvirus type 9 (EHV-9). Six 8-week-old SPF pigs were inoculated intranasally and four were inoculated orally with different doses of EHV-9, and observed for 6 days. Although neurological signs did not develop in any of the infected pigs, the six intranasally infected pigs and one of the orally infected pigs developed lesions of encephalitis consisting of neuronal necrosis, neuronophagia, and intranuclear inclusion bodies, distributed mainly in the rhinencephalon. EHV-9 antigen was localized in the necrotic neuronal cells and was closely associated with the presence of inclusion bodies. These findings clearly demonstrate that pigs are fully susceptible to EHV-9 infection following intranasal inoculation (but less so following oral inoculation), and that EHV-9 in pigs has a highly neurotropic nature. PMID- 11055875 TI - Mediastinal plasma cell tumor in a sheep. AB - An extramedullary plasmacytoma was found in a 10-year-old sheep. The tumor involved the mediastinum, where a 25 x 15 x 10-cm encapsulated mass was found. The lungs had multiple metastases ranging from 0.5 to 2 cm in diameter, and the portal vein contained a 10-cm-long mass. The cytologic and histopathologic analyses were consistent with a moderately differentiated plasmacytoma. The immunophenotype of the tumor cells was lambda light chain IgG+, CD79a-, and CD3-. Occasional granulomas were observed at the periphery of the mediastinal and pulmonary tumors. Microbiologic culture yielded growth of Corynebacterium from these granulomas. This is the first report of plasmacytoma in sheep. The tumor most likely arose from mediastinal lymph nodes and metastasized to the lungs and portal vein. PMID- 11055876 TI - Marginal siderosis and degenerative myelopathy: a manifestation of chronic subarachnoid hemorrhage in a horse with a myxopapillary ependymoma. AB - Marginal siderosis is recognized in humans as an uncommon clinicopathologic entity characterized by degeneration of neural tissue at the surface of the brain and spinal cord, in association with the accumulation of hemosiderin, and resulting from chronic subarachnoid hemorrhage. The sources of hemorrhage are various and include neoplasms, malformations, cysts, and vasculopathy. Marginal siderosis of the spinal cord due to a myxopapillary ependymoma was diagnosed in a 19-year-old Dutch Warm Blood horse with clinical signs of myelopathy. There is only one previous report of marginal siderosis in the veterinary literature, also in a horse with clinical myelopathy. PMID- 11055877 TI - Diagnosis of equine arteritis virus infection in two horses by using monoclonal antibody immunoperoxidase histochemistry on skin biopsies. AB - Two 5-year-old grade male horses presented with epiphora, rhinorrhea, conjunctival and nasal mucosal hyperemia, and dorsal and thoracic macropapular rash. Skin biopsies were collected from the affected areas, and serial sections were evaluated following hematoxylin and eosin and immunoperoxidase histochemistry staining by using a murine monoclonal antibody of the immunoglobulin G2A isotype recognizing the 30-kDa membrane protein of equine arteritis virus (EAV). In both horses, lesions consisted of mild to moderate diffuse superficial dermal edema and vasculitis with mild perivascular lymphocytic infiltrates, occasional endothelial hypertrophy, and single-cell necrosis of tunica media myocytes. Immunohistochemically, a few endothelial cells, myocytes, and pericytes containing intracytoplasmic EAV antigen were identified. Immunoperoxidase histochemistry of skin biopsies can be used as an ancillary test for the clinical diagnosis of equine viral arteritis in horses, especially when a cutaneous macropapular rash is evident. PMID- 11055878 TI - Ceroid-lipofuscinosis in a Cocker Spaniel dog. AB - A neurodegenerative storage disease identified as ceroid-lipofuscinosis by light, fluorescence, and electron microscopic examinations was diagnosed in a 4-year-old female Cocker Spaniel dog with progressive ataxia and proprioceptive deficits. Stored pigment was found within neurons of the brain and spinal cord and in smooth muscle cells of the urinary bladder and small muscular arteries. The microscopic findings resembled those found in six other cases of generalized ceroid-lipofuscinosis in this breed. However, the brown discoloration of the intestines, which was the major gross lesion observed in those cases, was not found. This is the first report of the disease in Argentina. PMID- 11055879 TI - Metastatic calcification in a dog attributable to ingestion of a tacalcitol ointment. AB - A 22-week-old 21-kg female Bernese Mountain Dog ingested a topical antipsoriatic preparation containing the synthetic vitamin D analog tacalcitol. The dog died after a history of lethargy, recumbency, paresis of the hindlimbs, increased rectal temperature, dyspnea, and hematemesis. Histologic examination revealed metastatic calcification in the kidneys, lungs, myocardium, brain, stomach, and tear glands. The appearance of soft tissue mineralization in multiple organs is consistent with hypercalcemia derived from excessive vitamin D uptake. Oral toxicity studies for tacalcitol in the dog are not available, but the present report emphasizes the extraordinary toxic risk of drugs containing this vitamin D analog to dogs. PMID- 11055880 TI - Oral focal epithelial hyperplasia in a howler monkey (Alouatta fusca). AB - Oral focal epithelial hyperplasia is a rare and seldom reported disease in animals and humans induced by a papillomavirus. The present report is the first description of this disease in a Neotropical primate, a howler monkey (Alouatta fusca). The diagnosis was based on gross and microscopic findings. The generic papillomavirus antigen was identified by immunohistochemistry and was found not to be related to any human papillomavirus DNA tested by in situ hybridization. This virus is probably a specific papillomavirus of the howler monkey (HMPV). PMID- 11055881 TI - Anti-plakin and desmoglein autoantibodies in a dog with pemphigus vulgaris. AB - Paraneoplastic pemphigus was suspected in a 14-year-old Labrador retriever because of mucocutaneous erosions, microscopic suprabasal acantholysis, and keratinocyte apoptosis. In this patient, circulating IgG autoantibodies recognized plakin (envoplakin, periplakin) and desmoglein (desmoglein-1 and -3) antigens. Necropsy, however, failed to confirm the concurrent existence of hematopoietic or solid neoplasia. The diagnosis of pemphigus vulgaris therefore was proposed. This study illustrates that such a combination of clinicopathologic lesions and plakin/desmoglein-specific autoantibodies is not restricted to canine paraneoplastic pemphigus but can also be detected in another form of suprabasal pemphigus. PMID- 11055882 TI - Pulmonary microcystic hamartoma in an adult dog. AB - Microcystic hamartoma was detected as a tumorlike mass in the left caudal lung lobe of a 12-year-old mixed-breed dog. Histologically, the mass was characterized by microcysts of various sizes that mimicked alveoli and were surrounded by thin fibrous septal tissue. However, unlike the adjacent lung parenchyma, bronchial or bronchiolar trees were absent, and the septal vascular channels were extremely underdeveloped. Immunohistochemically, the cells lining the microcysts were consistently positive for cytokeratin but not for vimentin, whereas the septal fibroblast-like cells were negative for cytokeratin and positive for vimentin. Electron microscopy confirmed that the microcysts were lined with a layer of type I and type II mature pneumocytes. This is the first description of the detailed morphologic features of microcystic hamartoma. PMID- 11055883 TI - Mucopolysaccharidosis VII in a cat. AB - Mucopolysaccharidosis VII was diagnosed in a domestic shorthair cat from California. The cat was small and had multiple abnormalities, including a small body disproportionate to the size of the skull, angular deformities of the ribs, abnormally short forelimbs, luxating patellas, generalized epiphyseal dysplasia involving the vertebrae and long bones, cuboidal vertebrae, pectus excavatum, subluxation of both hips, osteosclerosis of the tentorium cerebelli and left petrous temporal bone, tracheal hypoplasia, and corneal clouding. Beta glucuronidase activity was markedly decreased in peripheral blood leukocytes. The cat died at 21 months of age, and a complete necropsy was performed. Tissues were examined by light and transmission electron microscopy. Large clear, round vacuoles representing distended lysosomes were present in many epithelial and connective tissue cells, including fibrocytes, chondrocytes, smooth muscle cells, hepatocytes, astrocytes, and macrophages. PMID- 11055884 TI - Mammary adenocarcinoma in a male squirrel monkey (Saimiri sciureus). AB - A nodule was identified within the right mammary gland of a 16-year-old male squirrel monkey (Saimiri sciureus). The mass was excised and diagnosed as a mammary adenocarcinoma. The monkey developed congestive heart failure 1.5 years later and was euthanatized. At necropsy, a subcutaneous mass was found in the right axillary region. Histologically, the mass was identified as a lymph node whose architecture was effaced by neoplastic cells resembling those of the mammary tumor. Metastasis to internal organs was not observed. This is the first reported case of a mammary tumor in a New World primate and the only known case of mammary cancer in a male nonhuman primate. PMID- 11055885 TI - Tibial hemimelia, meningocele, and abdominal hernia in Shorthorn cattle. AB - Six genetically related Shorthorn calves were affected with the tibial hemimelia syndrome. The lesions included bilaterally malformed or absent tibia and abdominal hernia in all animals, a long shaggy haircoat, retained testicles in males, and meningocele in three animals. The malformations were similar to those described previously in Galloway calves. Pedigree analysis demonstrated a mechanism by which a recessive allele in a homozygous state could be responsible for the disorder. The condition in these calves was considered the result of a recurrence of a genetic mutation affecting a putative hemimelia locus. PMID- 11055886 TI - Cyclooxygenase-2 expression in inflammatory lung lesions of nonhuman primates. AB - Mammalian cells contain two related but unique isoforms of cyclooxygenase (COX-1 and COX-2). COX-1 is expressed constitutively in a majority of tissues and is involved in the production of prostaglandins (PGs) that modulate normal physiologic functions. COX-2 is inducible by various stimuli and is involved in the production of PGs that modulate physiologic events in development, cell growth, and inflammation. With the exception of peribronchial glands and chondrocytes of peribronchial cartilage, COX-2 is not detectable in the normal lung of nonhuman primates. We evaluated COX-2 expression by immunohistochemical methods in the inflammatory lesions of two cynomolgus monkeys (Macaca fascicularis) with acute severe pneumonia. Both monkeys exhibited acute severe bronchopneumonia; histologically, lung lesions were characterized by infiltration of large numbers of neutrophils and fewer macrophages, mild bronchial epithelial hyperplasia, and slight type-2 pneumocyte hyperplasia. In both monkeys, mild to marked COX-2 immunoreactivity was detected within the cytoplasm of macrophages, bronchial epithelial cells, type-2 pneumocytes, and endothelial cells of blood vessels. No COX-2 immunoreactivity was detectable in the neutrophils that constituted >90% of the inflammatory cells. These observations suggest that in acute inflammatory lung lesions in nonhuman primates 1) COX-2 is induced in the bronchial and alveolar epithelial cells, 2) macrophages are the primary inflammatory cells that exhibit COX-2, and 3) neutrophils do not express COX-2. PMID- 11055887 TI - New frontiers in deep vein thrombosis management. Symposium proceedings. Amsterdam, The Netherlands, July 4, 1998. PMID- 11055888 TI - Risk assessment of venous thromboembolism in medical patients. AB - Medical patients are at risk of venous thromboembolism (VTE). However, the extent of the risk is not well characterized. Risk stratification is an important part of effective thromboprophylactic management. The risks depend on the specific clinical risk factors in individual patients and include age, immobility, and prior history of VTE. Medical patient risk assessment models (RAMs) are not fully developed, but a number of specific risk factors have been characterized. Although at present prophylactic treatment is largely empirical, studies evaluating the effects of higher-dose regimens in at-risk patients are ongoing. Long-term prophylaxis may be of benefit to particular patient populations, including those with congenital and acquired aberrations in hemostasis. Published studies of low-molecular-weight heparins (LMWHs) show that these agents are at least as effective as low-dose unfractionated heparin (UFH), with a trend towards an improved safety profile. The results of ongoing studies will provide further insight into the risk of VTE in this important patient group. PMID- 11055889 TI - New therapeutic options in deep vein thrombosis prophylaxis. AB - Joint replacement surgery is complicated by a high rate of postoperative venous thromboembolism (VTE). Current thromboprophylactic approaches reduce the rate of VTE, but the incidence remains as high as 20% to 50%. Recombinant hirudins, such as desirudin and lepirudin, function by directly inhibiting thrombin, and are a new development in antithrombotic therapy. In two multicenter studies, desirudin was found to be superior to unfractionated heparin (UFH) in the prevention of deep vein thrombosis (DVT) after total hip alloplasty. A further trial of more than 2,000 patients undergoing elective hip replacement compared the thromboprophylactic efficacy of desirudin versus the low-molecular-weight heparin (LMWH) enoxaparin. Desirudin was more effective than LMWH in providing effective prophylaxis, and maintained superiority in patients with additional risk. Desirudin was shown to be equally safe and did not require laboratory monitoring. Desirudin (15 mg twice daily) is an efficient therapy for DVT prevention in hip alloplasty patients at additional risk. PMID- 11055890 TI - Assessment of treatment strategy in high-risk surgical patients. AB - The risk of postoperative venous thromboembolism (VTE) is dependent on the type of surgical procedure and specific characteristics of the patient. Patients with intrinsic factors placing them at higher risk of VTE remain at high risk, even if the procedural risk is low. The identification of factors such as age, obesity, and malignant disease, has made it possible to make a preoperative assessment of risk and ensures that the most appropriate thromboprophylactic regimen is prescribed. Despite the widespread use of prophylaxis, the incidence of VTE is still significant. The investigation and development of new pharmacologic agents, such as hirudins and their recombinant derivatives, may lead to better protection for patients at high or very high risk of VTE. PMID- 11055891 TI - Thromboprophylaxis in neurosurgical patients. AB - Neurosurgical patients constitute one of the highest risk groups for postoperative thromboembolic complications. Physical methods of thromboprophylaxis have been successful in reducing the incidence of postoperative deep vein thrombosis (DVT) but the residual incidence remains considerable. Many neurosurgeons are reluctant to use perioperative anticoagulant prophylaxis, despite its proven success in reducing DVT rates, because of the potentially serious consequences of even a small intracranial bleed. Recent studies have indicated that a combination of graduated compression stockings (GCS) and low-molecular-weight heparin (LMWH), started in the postoperative period, significantly reduces the incidence of DVT compared with GCS alone. Postoperative regimens avoid the risk of surgical hemorrhage and appear to offer increased protection for this group of patients. PMID- 11055892 TI - Venous thromboembolic disease management of the nonsurgical moderate- and high risk patient. AB - Venous thromboembolism (VTE) is at least as common among medical as in surgical patients, and fatal pulmonary embolism (PE) occurs most commonly in medical patients. As in surgical patients, risk factors and high- and low-risk groups have been identified, but no useful risk assessment models (RAMs) have yet been developed for medical patients. The incidence of VTE varies widely, but a history of previous VTE, age greater than 40 years, obesity, and prolonged immobilization are all associated with an increase In risk. Stroke, spinal cord injury, and cancer patients are at particular risk of VTE. Low-dose unfractionated heparin (UFH), low-molecular-weight heparins (LMWHs), antiplatelet agents, and warfarin reduce the incidence of VTE. Current standards of thromboprophylaxis vary with indication. However, utilization is frequently low, even in high-risk patients. Effective prophylaxis must be tailored to the individual patient, taking into account both the history of the patient and the disease. PMID- 11055893 TI - Management of thromboembolism in the outpatient setting. AB - Low-molecular-weight heparins (LMWHs) are suitable for self-administration at home, because they have a predictable anticoagulant effect following subcutaneous injection and do not require laboratory monitoring. Clinical trials evaluating the safety and efficacy of LMWHs in the outpatient setting for the prevention of deep vein thrombosis (DVT) after orthopedic surgery and for the treatment of established DVT are reviewed. Extended LMWH prophylaxis reduces the incidence of venographically detected DVT by approximately 50%. Medical practice relies heavily on clinical diagnosis of DVT, for which both sensitivity and specificity are poor. It is uncertain how the results of research trials on DVT prevention based on venography relate to ordinary practice. In the treatment of established DVT, there was no significant difference between outpatient management with LMWH and inpatient treatment with unfractionated heparin (UFH). However, outpatient management offered a considerable reduction in resource usage, with associated cost savings. PMID- 11055895 TI - Molecular characterization of 3-phosphoglycerate dehydrogenase deficiency--a neurometabolic disorder associated with reduced L-serine biosynthesis. AB - 3-phosphoglycerate dehydrogenase (PHGDH) deficiency is a disorder of L-serine biosynthesis that is characterized by congenital microcephaly, psychomotor retardation, and seizures. To investigate the molecular basis for this disorder, the PHGDH mRNA sequence was characterized, and six patients from four families were analyzed for sequence variations. Five patients from three different families were homozygous for a single nucleotide substitution predicted to change valine at position 490 to methionine. The sixth patient was homozygous for a valine to methionine substitution at position 425; both mutations are located in the carboxyterminal part of PHGDH. In vitro expression of these mutant proteins resulted in significant reduction of PHGDH enzyme activities. RNA-blot analysis indicated abundant expression of PHGDH in adult and fetal brain tissue. Taken together with the severe neurological impairment in our patients, the data presented in this paper suggest an important role for PHGDH activity and L-serine biosynthesis in the metabolism, development, and function of the central nervous system. PMID- 11055896 TI - Distinct missense mutations of the FGFR3 lys650 codon modulate receptor kinase activation and the severity of the skeletal dysplasia phenotype. AB - The fibroblast growth factor-receptor 3 (FGFR3) Lys650 codon is located within a critical region of the tyrosine kinase-domain activation loop. Two missense mutations in this codon are known to result in strong constitutive activation of the FGFR3 tyrosine kinase and cause three different skeletal dysplasia syndromes thanatophoric dysplasia type II (TD2) (A1948G [Lys650Glu]) and SADDAN (severe achondroplasia with developmental delay and acanthosis nigricans) syndrome and thanatophoric dysplasia type I (TD1) (both due to A1949T [Lys650Met]). Other mutations within the FGFR3 tyrosine kinase domain (e.g., C1620A or C1620G [both resulting in Asn540Lys]) are known to cause hypochondroplasia, a relatively common but milder skeletal dysplasia. In 90 individuals with suspected clinical diagnoses of hypochondroplasia who do not have Asn540Lys mutations, we screened for mutations, in FGFR3 exon 15, that would disrupt a unique BbsI restriction site that includes the Lys650 codon. We report here the discovery of three novel mutations (G1950T and G1950C [both resulting in Lys650Asn] and A1948C [Lys650Gln]) occurring in six individuals from five families. Several physical and radiological features of these individuals were significantly milder than those in individuals with the Asn540Lys mutations. The Lys650Asn/Gln mutations result in constitutive activation of the FGFR3 tyrosine kinase but to a lesser degree than that observed with the Lys540Glu and Lys650Met mutations. These results demonstrate that different amino acid substitutions at the FGFR3 Lys650 codon can result in several different skeletal dysplasia phenotypes. PMID- 11055897 TI - Active intestinal chloride secretion in human carriers of cystic fibrosis mutations: an evaluation of the hypothesis that heterozygotes have subnormal active intestinal chloride secretion. AB - To explain the very high frequency of cystic fibrosis (CF) mutations in most populations of European descent, it has been proposed that CF heterozygotes have a survival advantage when infected with Vibrio cholerae or Escherichia coli, the toxins of which induce diarrhea by stimulation of active intestinal chloride secretion. Two assumptions underlie this hypothesis: (1) chloride conductance by the CF transmembrane conductance regulator (CFTR) is the rate-limiting step for active intestinal chloride secretion at all levels of expression, from approximately zero in patients with CF to normal levels in people who are not carriers of a mutation; and (2) heterozygotes have smaller amounts of functional intestinal CFTR than do people who are not carriers, and heterozygotes therefore secrete less chloride when exposed to secretagogues. The authors used an intestinal perfusion technique to measure in vivo basal and prostaglandin stimulated jejunal chloride secretion in normal subjects, CF heterozygotes, and patients with CF. Patients with CF had essentially no active chloride secretion in the basal state, and secretion was not stimulated by a prostaglandin analogue. However, CF heterozygotes secreted chloride at the same rate as did people without a CF mutation. If heterozygotes are assumed to have less-than-normal intestinal CFTR function, these results mean that CFTR expression is not rate limiting for active chloride secretion in heterozygotes. The results do not support the theory that the very high frequency of CF mutations is due to a survival advantage that is conferred on heterozygotes who contract diarrheal illnesses mediated by intestinal hypersecretion of chloride. PMID- 11055898 TI - Diagnostic testing for Rett syndrome by DHPLC and direct sequencing analysis of the MECP2 gene: identification of several novel mutations and polymorphisms. AB - Rett syndrome (RTT) is an X-linked dominant neurodevelopmental disorder affecting 1/10,000-15,000 girls. The disease-causing gene was identified as MECP2 on chromosome Xq28, and mutations have been found in approximately 80% of patients diagnosed with RTT. Numerous mutations have been identified in de novo and rare familial cases, and they occur primarily in the methyl-CpG-binding and transcriptional-repression domains of MeCP2. Our first diagnostic strategy used bidirectional sequencing of the entire MECP2 coding region. Subsequently, we implemented a two-tiered strategy that used denaturing high-performance liquid chromatography (DHPLC) for initial screening of nucleotide variants, followed by confirmatory sequencing analysis. If a definite mutation was not identified, then the entire MECP2 coding region was sequenced, to reduce the risk of false negatives. Collectively, we tested 228 unrelated female patients with a diagnosis of possible (209) or classic (19) RTT and found MECP2 mutations in 83 (40%) of 209 and 16 (84%) of 19 of the patients, respectively. Thirty-two different mutations were identified (8 missense, 9 nonsense, 1 splice site, and 14 frameshifts), of which 12 are novel and 9 recurrent in unrelated patients. Seven unclassified variants and eight polymorphisms were detected in 228 probands. Interestingly, we found that T203M, previously reported as a missense mutation in an autistic patient, is actually a benign polymorphism, according to parental analysis performed in a second case identified in this study. These findings highlight the complexities of missense variant interpretation and emphasize the importance of parental DNA analysis for establishing an etiologic relation between a possible mutation and disease. Overall, we found a 98.8% concordance rate between DHPLC and sequence analyses. One mutation initially missed by the DHPLC screening was identified by sequencing. Modified conditions subsequently enabled its detection, underscoring the need for multiple optimized conditions for DHPLC analysis. We conclude that this two-tiered approach provides a sensitive, robust, and efficient strategy for RTT molecular diagnosis. PMID- 11055899 TI - Genetic control of resistance to the sterol 14alpha-demethylase inhibitor fungicide prochloraz in the cereal eyespot pathogen Tapesia yallundae. AB - Sexual crosses were used to determine the genetic basis of resistance to the sterol 14 alpha-demethylase inhibitor fungicide prochloraz in the cereal eyespot pathogen Tapesia yallundae. Three different crosses between sensitive parental strains (22-432 and 22-433 [the concentration required to inhibit growth by 50% (IG(50)) for each was 10 g(-1) tdh- and/or trh-positive V. parahaemolyticus in environmental oysters should be considered extraordinary. PMID- 11055907 TI - Isolation and use of a homologous histone H4 promoter and a ribosomal DNA region in a transformation vector for the oil-producing fungus Mortierella alpina. AB - Mortierella alpina was transformed successfully to hygromycin B resistance by using a homologous histone H4 promoter to drive gene expression and a homologous ribosomal DNA region to promote chromosomal integration. This is the first description of transformation in this commercially important oleaginous organism. Two pairs of histone H3 and H4 genes were isolated from this fungus. Each pair consisted of one histone H3 gene and one histone H4 gene, transcribed divergently from an intergenic promoter region. The pairs of encoded histone H3 or H4 proteins were identical in amino acid sequence. At the DNA level, each histone H3 or H4 open reading frame showed 97 to 99% identity to its counterpart but the noncoding regions had little sequence identity. Unlike the histone genes from other filamentous fungi, all four M. alpina genes lacked introns. During normal vegetative growth, transcripts from the two histone H4 genes were produced at approximately the same level, indicating that either histone H4 promoter could be used in transformation vectors. The generation of stable, hygromycin B-resistant transformants required the incorporation of a homologous ribosomal DNA region into the transformation vector to promote chromosomal integration. PMID- 11055908 TI - Key aromatic-ring-cleaving enzyme, protocatechuate 3,4-dioxygenase, in the ecologically important marine Roseobacter lineage. AB - Aromatic compound degradation in six bacteria representing an ecologically important marine taxon of the alpha-proteobacteria was investigated. Initial screens suggested that isolates in the Roseobacter lineage can degrade aromatic compounds via the beta-ketoadipate pathway, a catabolic route that has been well characterized in soil microbes. Six Roseobacter isolates were screened for the presence of protocatechuate 3,4-dioxygenase, a key enzyme in the beta-ketoadipate pathway. All six isolates were capable of growth on at least three of the eight aromatic monomers presented (anthranilate, benzoate, p-hydroxybenzoate, salicylate, vanillate, ferulate, protocatechuate, and coumarate). Four of the Roseobacter group isolates had inducible protocatechuate 3, 4-dioxygenase activity in cell extracts when grown on p-hydroxybenzoate. The pcaGH genes encoding this ring cleavage enzyme were cloned and sequenced from two isolates, Sagittula stellata E-37 and isolate Y3F, and in both cases the genes could be expressed in Escherichia coli to yield dioxygenase activity. Additional genes involved in the protocatechuate branch of the beta-ketoadipate pathway (pcaC, pcaQ, and pobA) were found to cluster with pcaGH in these two isolates. Pairwise sequence analysis of the pca genes revealed greater similarity between the two Roseobacter group isolates than between genes from either Roseobacter strain and soil bacteria. A degenerate PCR primer set targeting a conserved region within PcaH successfully amplified a fragment of pcaH from two additional Roseobacter group isolates, and Southern hybridization indicated the presence of pcaH in the remaining two isolates. This evidence of protocatechuate 3, 4-dioxygenase and the beta-ketoadipate pathway was found in all six Roseobacter isolates, suggesting widespread abilities to degrade aromatic compounds in this marine lineage. PMID- 11055909 TI - Rhizosphere competitiveness of trichloroethylene-degrading, poplar-colonizing recombinant bacteria. AB - Indigenous bacteria from poplar tree (Populus canadensis var. eugenei 'Imperial Carolina') and southern California shrub rhizospheres, as well as two tree colonizing Rhizobium strains (ATCC 10320 and ATCC 35645), were engineered to express constitutively and stably toluene o-monooxygenase (TOM) from Burkholderia cepacia G4 by integrating the tom locus into the chromosome. The poplar and Rhizobium recombinant bacteria degraded trichloroethylene at a rate of 0.8 to 2.1 nmol/min/mg of protein and were competitive against the unengineered hosts in wheat and barley rhizospheres for 1 month (colonization occurred at a level of 1.0 x 10(5) to 23 x 10(5) CFU/cm of root). In addition, six of these recombinants colonized poplar roots stably and competitively with populations as large as 79% +/- 12% of all rhizosphere bacteria after 28 days (0.2 x 10(5) to 31 x 10(5) CFU/cm of root). Furthermore, five of the most competitive poplar recombinants (e.g., Pb3-1 and Pb5-1, which were identified as Pseudomonas sp. strain PsK recombinants) retained the ability to express TOM for 29 days as 100% +/- 0% of the recombinants detected in the poplar rhizosphere expressed TOM constitutively. PMID- 11055910 TI - Attachment of Escherichia coli O157:H7 to the surfaces and internal structures of apples as detected by confocal scanning laser microscopy. AB - Confocal scanning laser microscopy (CSLM) was used to demonstrate the attachment of Escherichia coli O157:H7 transformed with a plasmid encoding for green fluorescent protein (GFP) to the surface and within the internal structures of nonwaxed Red Delicious cv. apples. Apples at 2 or 25 degrees C were inoculated with an E. coli O157:H7 cell suspension at 2 or 25 degrees C. The effect of a negative temperature differential (cold inoculum, warm apple), a positive differential (warm inoculum, cold apple), and no differential (warm inoculum, warm apple), in combination with a pressure differential (atmospheric versus 10,130 Pa), on the attachment and infiltration of cells was determined. CSLM stereo images of external surfaces of apples subjected to all combinations of test parameters showed preferential cellular attachment to discontinuities in the waxy cuticle on the surface and to damaged tissue surrounding puncture wounds, where the pathogen was observed at depths up to 70 microm below the skin surface. Attachment to lenticels was sporadic but was occasionally observed at depths of up to 40 microm. Infiltration through the floral tube and attachment to seeds, cartilaginous pericarp, and internal trichomes were observed in all apples examined, regardless of temperature differential during inoculation. The pressure differential had no effect on infiltration or attachment of E. coli O157:H7. Image analysis to count cells at various depths within tissues was used to quantitatively compare the extent of infiltration into various apple structures as well as the effects of the temperature differential. Puncture wounds harbored greater numbers of the pathogen at greater depths than did other sites examined. Attachment or infiltration of cells was greater on the intact skin and in lenticels, russet areas, and the floral tube of apples inoculated under a negative temperature differential compared to those inoculated under no temperature differential. The results suggest that E. coli O157:H7 attached to internal core structures or within tissues of apples may evade decontamination treatments. Interventions designed to deliver disinfectants to these locations or to remove viable cells of E. coli O157:H7 and other pathogens from apples by other means need to be developed and validated. PMID- 11055911 TI - Characterization of pRGO1, a plasmid from Propionibacterium acidipropionici, and its use for development of a host-vector system in propionibacteria. AB - The complete nucleotide sequence of pRGO1, a cryptic plasmid from Propionibacterium acidipropionici E214, was determined. pRGO1 is 6, 868 bp long, and its G+C content is 65.0%. Frame analysis of the sequence revealed six open reading frames, which were designated Orf1 to Orf6. The deduced amino acid sequences of Orf1 and Orf2 showed extensive similarities to an initiator of plasmid replication, the Rep protein, of various plasmids of gram-positive bacteria. The amino acid sequence of the putative translation product of orf3 exhibited a high degree of similarity to the amino acid sequences of DNA invertase in several bacteria. For the putative translation products of orf4, orf5, and orf6, on the other hand, no homologous sequences were found. The function of these open reading frames was studied by deletion analysis. A shuttle vector, pPK705, was constructed for shuttling between Escherichia coli and a Propionibacterium strain containing orf1 (repA), orf2 (repB), orf5, and orf6 from pRGO1, pUC18, and the hygromycin B-resistant gene as a drug marker. Shuttle vector pPK705 successfully transformed Propionibacterium freudenreichii subsp. shermanii IFO12426 by electroporation at an efficiency of 8 x 10(6) CFU/microg of DNA under optimized conditions. Transformation of various species of propionibacteria with pPK705 was also performed at efficiencies of about 10(4) to 10(7) CFU/microg of DNA. The vector was stably maintained in strains of P. freudenreichii subsp. shermanii, P. freudenreichii, P. pentosaceum, and P. freudenreichii subsp. freudenreichii grown under nonselective conditions. Successful manipulation of a host-vector system in propionibacteria should facilitate genetic studies and lead to creation of genes that are useful industrially. PMID- 11055912 TI - Identification and characterization of an ATP binding cassette L-carnitine transporter in Listeria monocytogenes. AB - We identified an operon in Listeria monocytogenes EGD with high levels of sequence similarity to the operons encoding the OpuC and OpuB compatible solute transporters from Bacillus subtilis, which are members of the ATP binding cassette (ABC) substrate binding protein-dependent transporter superfamily. The operon, designated opuC, consists of four genes which are predicted to encode an ATP binding protein (OpuCA), an extracellular substrate binding protein (OpuCC), and two membrane-associated proteins presumed to form the permease (OpuCB and OpuCD). The operon is preceded by a potential SigB-dependent promoter. An opuC defective mutant was generated by the insertional inactivation of the opuCA gene. The mutant was impaired for growth at high osmolarity in brain heart infusion broth and failed to grow in a defined medium. Supplementation of the defined medium with peptone restored the growth of the mutant in this medium. The mutant was found to accumulate the compatible solutes glycine betaine and choline to same extent as the parent strain but was defective in the uptake of L-carnitine. We conclude that the opuC operon in L. monocytogenes encodes an ABC compatible solute transporter which is capable of transporting L-carnitine and which plays an important role in osmoregulation in this pathogen. PMID- 11055913 TI - Denaturing gradient gel electrophoresis analysis of 16S ribosomal DNA amplicons to monitor changes in fecal bacterial populations of weaning pigs after introduction of Lactobacillus reuteri strain MM53. AB - The diversity and stability of the fecal bacterial microbiota in weaning pigs was studied after introduction of an exogenous Lactobacillus reuteri strain, MM53, using a combination of cultivation and techniques based on genes encoding 16S rRNA (16S rDNA). Piglets (n = 9) were assigned to three treatment groups (control, daily dosed, and 4th-day dosed), and fresh fecal samples were collected daily. Dosed animals received 2.5 x 10(10) CFU of antibiotic-resistant L. reuteri MM53 daily or every 4th day. Mean Lactobacillus counts for the three groups ranged from 1 x 10(9) to 4 x 10(9) CFU/g of feces. Enumeration of strain L. reuteri MM53 on MRS agar (Difco) plates containing streptomycin and rifampin showed that the introduced strain fluctuated between 8 x 10(3) and 5 x 10(6) CFU/g of feces in the two dosed groups. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA fragments, with primers specific for variable regions 1 and 3 (V1 and V3), was used to profile complexity of fecal bacterial populations. Analysis of DGGE banding profiles indicated that each individual maintained a unique fecal bacterial population that was stable over time, suggesting a strong host influence. In addition, individual DGGE patterns could be separated into distinct time-dependent clusters. Primers designed specifically to restrict DGGE analysis to a select group of lactobacilli allowed examination of interspecies relationships and abundance. Based on relative band migration distance and sequence determination, L. reuteri was distinguishable within the V1 region 16S rDNA gene patterns. Daily fluctuations in specific bands within these profiles were observed, which revealed an antagonistic relationship between L. reuteri MM53 (band V1-3) and another indigenous Lactobacillus assemblage (band V1 6). PMID- 11055914 TI - adhA in Aspergillus parasiticus is involved in conversion of 5'-hydroxyaverantin to averufin. AB - Two routes for the conversion of 5'-hydroxyaverantin (HAVN) to averufin (AVF) in the synthesis of aflatoxin have been proposed. One involves the dehydration of HAVN to the lactone averufanin (AVNN), which is then oxidized to AVF. Another requires dehydrogenation of HAVN to 5'-ketoaverantin, the open-chain form of AVF, which then cyclizes spontaneously to AVF. We isolated a gene, adhA, from the aflatoxin gene cluster of Aspergillus parasiticus SU-1. The deduced ADHA amino acid sequence contained two conserved motifs found in short-chain alcohol dehydrogenases-a glycine-rich loop (GXXXGXG) that is necessary for interaction with NAD(+)-NADP(+), and the motif YXXXK, which is found at the active site. A. parasiticus SU-1, which produces aflatoxins, has two copies of adhA (adhA1), whereas A. parasiticus SRRC 2043, a strain that accumulates O methylsterigmatocystin (OMST), has only one copy. Disruption of adhA in SRRC 2043 resulted in a strain that accumulates predominantly HAVN. This result suggests that ADHA is involved in the dehydrogenation of HAVN to AVF. Those adhA disruptants that still made small amounts of OMST also accumulated other metabolites, including AVNN, after prolonged culture. PMID- 11055915 TI - Characterization of Cryptosporidium parvum by matrix-assisted laser desorption ionization-time of flight mass spectrometry. AB - Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was used to investigate whole and freeze-thawed Cryptosporidium parvum oocysts. Whole oocysts revealed some mass spectral features. Reproducible patterns of spectral markers and increased sensitivity were obtained after the oocysts were lysed with a freeze-thaw procedure. Spectral-marker patterns for C. parvum were distinguishable from those obtained for Cryptosporidium muris. One spectral marker appears specific for the genus, while others appear specific at the species level. Three different C. parvum lots were investigated, and similar spectral markers were observed in each. Disinfection of the oocysts reduced and/or eliminated the patterns of spectral markers. PMID- 11055916 TI - Detection and identification of decay fungi in spruce wood by restriction fragment length polymorphism analysis of amplified genes encoding rRNA. AB - We have developed a DNA-based assay to reliably detect brown rot and white rot fungi in wood at different stages of decay. DNA, isolated by a series of CTAB (cetyltrimethylammonium bromide) and organic extractions, was amplified by the PCR using published universal primers and basidiomycete-specific primers derived from ribosomal DNA sequences. We surveyed 14 species of wood-decaying basidiomycetes (brown-rot and white-rot fungi), as well as 25 species of wood inhabiting ascomycetes (pathogens, endophytes, and saprophytes). DNA was isolated from pure cultures of these fungi and also from spruce wood blocks colonized by individual isolates of wood decay basidiomycetes or wood-inhabiting ascomycetes. The primer pair ITS1-F (specific for higher fungi) and ITS4 (universal primer) amplified the internal transcribed spacer region from both ascomycetes and basidiomycetes from both pure culture and wood, as expected. The primer pair ITS1 F (specific for higher fungi) and ITS4-B (specific for basidiomycetes) was shown to reliably detect the presence of wood decay basidiomycetes in both pure culture and wood; ascomycetes were not detected by this primer pair. We detected the presence of decay fungi in wood by PCR before measurable weight loss had occurred to the wood. Basidiomycetes were identified to the species level by restriction fragment length polymorphisms of the internal transcribed spacer region. PMID- 11055917 TI - Ingested blood contributes to the specificity of the symbiosis of Aeromonas veronii biovar sobria and Hirudo medicinalis, the medicinal leech. AB - Hirudo medicinalis, the medicinal leech, usually carries in its digestive tract a pure culture of Aeromonas veronii bv. sobria. Such specificity is unusual for digestive tracts that are normally colonized by a complex microbial consortium. Important questions for the symbiotic interaction and for the medical application after microvascular surgery are whether other bacteria can proliferate or at least persist in the digestive tract of H. medicinalis and what factors contribute to the reported specificity. Using a colonization assay, we were able to compare experimentally the ability of clinical isolates and of a symbiotic strain to colonize H. medicinalis. The symbiotic A. veronii bv. sobria strain proliferated well and persisted for at least 7 days inside the digestive tract. In contrast, the proliferation of Pseudomonas aeruginosa and Staphylococcus aureus was inhibited inside the animal compared to growth in the in vitro control, indicating that the ingested blood was modified within the digestive tract. However, both strains were able to persist in the digestive tract for at least 7 days. For an Escherichia coli strain, the viable counts decreased approximately 1, 000-fold within 42 h. The decrease of viable E. coli could be prevented by interfering with the activation of the membrane-attack complex of the complement system that is present in blood. This suggests that the membrane attack complex remained active inside H. medicinalis and prevented the proliferation of sensitive bacteria. Thus, antimicrobial properties of the ingested vertebrate blood contribute to the specificity of the A. veronii-H. medicinalis symbiosis, in addition to modifications of the blood inside the digestive tract of H. medicinalis. PMID- 11055918 TI - Transformation of 2,4,6-trinitrotoluene by purified xenobiotic reductase B from Pseudomonas fluorescens I-C. AB - The enzymatic transformation of 2,4,6-trinitrotoluene (TNT) by purified XenB, an NADPH-dependent flavoprotein oxidoreductase from Pseudomonas fluorescens I-C, was evaluated by using natural abundance and [U-(14)C]TNT preparations. XenB catalyzed the reduction of TNT either by hydride addition to the aromatic ring or by nitro group reduction, with the accumulation of various tautomers of the protonated dihydride-Meisenheimer complex of TNT, 2-hydroxylamino-4,6 dinitrotoluene, and 4-hydroxylamino-2, 6-dinitrotoluene. Subsequent reactions of these metabolites were nonenzymatic and resulted in predominant formation of at least three dimers with an anionic m/z of 376 as determined by negative-mode electrospray ionization mass spectrometry and the release of approximately 0.5 mol of nitrite per mol of TNT consumed. The extents of the initial enzymatic reactions were similar in the presence and in the absence of O(2), but the dimerization reaction and the release of nitrite were favored under aerobic conditions or under anaerobic conditions in the presence of NADP(+). Reactions of chemically and enzymatically synthesized and high-pressure liquid chromatography purified TNT metabolites showed that both a hydroxylamino-dinitrotoluene isomer and a tautomer of the protonated dihydride-Meisenheimer complex of TNT were required precursors for the dimerization and nitrite release reactions. The m/z 376 dimers also reacted with either dansyl chloride or N-1 naphthylethylenediamine HCl, providing evidence for an aryl amine functional group. In combination, the experimental results are consistent with assigning the chemical structures of the m/z 376 species to various isomers of amino-dimethyl tetranitrobiphenyl. A mechanism for the formation of these proposed TNT metabolites is presented, and the potential enzymatic and environmental significance of their formation is discussed. PMID- 11055919 TI - Isolation and characterization of a Helicobacter sp. from the gastric mucosa of dolphins, Lagenorhynchus acutus and Delphinus delphis. AB - Gastric ulcerations in dolphins have been reported for decades. Some of these lesions were associated with parasitic infections. However, cases of nonparasitic gastric ulcers with no clearly defined etiology also have been reported in wild and captive dolphins. Considerable speculation exists as to whether dolphins have Helicobacter-associated gastritis and peptic ulcer disease. The stomachs of seven stranded Atlantic white-sided dolphins, Lagenorhynchus acutus, and 1 common dolphin, Delphinus delphis, were assessed for the presence of Helicobacter species. Novel Helicobacter species were identified by culture in the gastric mucosa of two of the eight dolphins studied and by PCR in seven of the eight dolphins. The gram-negative organisms were urease, catalase, and oxidase positive. Spiral to fusiform bacteria were detected in gastric mucosa by Warthin Starry staining. Histopathology revealed mild to moderate diffuse lymphoplasmacytic gastritis within the superficial mucosa of the main stomach. The pyloric stomach was less inflamed, and bacteria did not extend deep into the glands. The lesions parallel those observed in Helicobacter pylori-infected humans. Bacteria from two dolphins classified by 16S rRNA analysis clustered with gastric helicobacters and represent a novel Helicobacter sp. most closely related to H. pylori. These findings suggest that a novel Helicobacter sp. may play a role in the etiopathogenesis of gastritis and gastric ulcers in dolphins. To our knowledge this represents the first isolation and characterization of a novel Helicobacter sp. from a marine mammal and emphasizes the wide host distribution and pathogenic potential of this increasingly important genus. PMID- 11055920 TI - Identification of a universally primed-PCR-derived sequence-characterized amplified region marker for an antagonistic strain of Clonostachys rosea and development of a strain-specific PCR detection assay. AB - We developed a PCR detection method that selectively recognizes a single biological control agent and demonstrated that universally primed PCR (UP-PCR) can identify strain-specific markers. Antagonistic strains of Clonostachys rosea (syn. Gliocladium roseum) were screened by UP-PCR, and a strain-specific marker was identified for strain GR5. No significant sequence homology was found between this marker and any other sequences in the databases. Southern blot analysis of the PCR product revealed that the marker represented a single-copy sequence specific for strain GR5. The marker was converted into a sequence-characterized amplified region (SCAR), and a specific PCR primer pair was designed. Eighty-two strains, isolated primarily from Danish soils, and 31 soil samples, originating from different localities, were tested, and this specificity was confirmed. Two strains responded to the SCAR primers under suboptimal PCR conditions, and the amplified sequences from these strains were similar, but not identical, to the GR5 marker. Soil assays in which total DNA was extracted from GR5-infested and noninoculated field soils showed that the SCAR primers could detect GR5 in a pool of mixed DNA and that no other soil microorganisms present contained sequences amplified by the primers. The assay developed will be useful for monitoring biological control agents released into natural field soil. PMID- 11055921 TI - Cloning and random mutagenesis of the Erwinia herbicola tyrR gene for high-level expression of tyrosine phenol-lyase. AB - Tyrosine phenol-lyase (Tpl), which can synthesize 3, 4-dihydroxyphenylalanine from pyruvate, ammonia, and catechol, is a tyrosine-inducible enzyme. Previous studies demonstrated that the tpl promoter of Erwinia herbicola is activated by the TyrR protein of Escherichia coli. In an attempt to create a high-Tpl expressing strain, we cloned the tyrR gene of E. herbicola and then randomly mutagenized it. Mutant TyrR proteins with enhanced ability to activate tpl were screened for by use of the lac reporter system in E. coli. The most increased transcription of tpl was observed for the strain with the mutant tyrR allele involving amino acid substitutions of alanine, cysteine, and glycine for valine 67, tyrosine-72, and glutamate-201, respectively. A tyrR-deficient derivative of E. herbicola was constructed and transformed with a plasmid carrying the mutant tyrR allele (V67A Y72C E201G substitutions). The resultant strain expressed Tpl without the addition of tyrosine to the medium and produced as much of it as was produced by the wild-type strain grown under tyrosine-induced conditions. The regulatory properties of the mutant TyrR(V67A), TyrR(Y72C), TyrR(E201G), and TyrR(V67A Y72C E201G) proteins were examined in vivo. Interestingly, as opposed to the wild-type TyrR protein, the mutant TyrR(V67A) protein had a repressive effect on the tyrP promoter in the presence of phenylalanine as the coeffector. PMID- 11055922 TI - Streptococcus thermophilus cell wall-anchored proteinase: release, purification, and biochemical and genetic characterization. AB - Streptococcus thermophilus CNRZ 385 expresses a cell envelope proteinase (PrtS), which is characterized in the present work, both at the biochemical and genetic levels. Since PrtS is resistant to most classical methods of extraction from the cell envelopes, we developed a three-step process based on loosening of the cell wall by cultivation of the cells in the presence of glycine (20 mM), mechanical disruption (with alumina powder), and enzymatic treatment (lysozyme). The pure enzyme is a serine proteinase highly activated by Ca(2+) ions. Its activity was optimal at 37 degrees C and pH 7.5 with acetyl-Ala-Ala-Pro-Phe-paranitroanilide as substrate. The study of the hydrolysis of the chromogenic and casein substrates indicated that PrtS presented an intermediate specificity between the most divergent types of cell envelope proteinases from lactococci, known as the PI and PIII types. This result was confirmed by the sequence determination of the regions involved in substrate specificity, which were a mix between those of PI and PIII types, and also had unique residues. Sequence analysis of the PrtS encoding gene revealed that PrtS is a member of the subtilase family. It is a multidomain protein which is maturated and tightly anchored to the cell wall via a mechanism involving an LPXTG motif. PrtS bears similarities to cell envelope proteinases from pyogenic streptococci (C5a peptidase and cell surface proteinase) and lactic acid bacteria (PrtP, PrtH, and PrtB). The highest homologies were found with streptococcal proteinases which lack, as PrtS, one domain (the B domain) present in cell envelope proteinases from all other lactic acid bacteria. PMID- 11055923 TI - Molecular epidemiological survey of Listeria monocytogenes in seafoods and seafood-processing plants. AB - To evaluate the role of seafoods in the epidemiology of human listeriosis and the role of the processing environment as a source of Listeria monocytogenes in seafood products, 305 L. monocytogenes isolates were characterized by multilocus enzyme electrophoresis using 21 genetic loci and restriction enzyme analysis of total DNA. Forty-four isolates were recovered from patients in Norway; 93 were isolated from seafoods, seafood-processing environments, and seawater from 55 different producers; and the remaining 168 isolates originated from six seafood processing plants and one transport terminal examined in detail for L. monocytogenes. The patient isolates fell into 11 electrophoretic types, with four of them being responsible for 77% of the listeriosis cases in 1992 to 1996. Isolates from Norwegian seafoods and processing environments showed great genetic diversity, indicating that seafoods and seafood-processing environments do not offer a niche for specific L. monocytogenes strains. On the other hand, isolates from individual processing plants were genetically more homogenous, showing that plants are likely to be colonized with specific subclones of L. monocytogenes. The isolation of identical subclones of L. monocytogenes from both human patients and seafoods, including ready-to-eat products, suggests that such products may have been possible sources for listeriosis cases in Norway. PMID- 11055924 TI - Genetic diversity and dynamics of Sinorhizobium meliloti populations nodulating different alfalfa cultivars in Italian soils. AB - We analyzed the genetic diversity of 531 Sinorhizobium meliloti strains isolated from nodules of Medicago sativa cultivars in two different Italian soils during 4 years of plant growth. The isolates were analyzed for DNA polymorphism with the random amplified polymorphic DNA method. The populations showed a high level of genetic polymorphism distributed throughout all the isolates, with 440 different haplotypes. Analysis of molecular variance allowed us to relate the genetic structure of the symbiotic population to various factors, including soil type, alfalfa cultivar, individual plants within a cultivar, and time. Some of these factors significantly affected the genetic structure of the population, and their relative influence changed with time. At the beginning of the experiment, the soil of origin and, even more, the cultivar significantly influenced the distribution of genetic variability of S. meliloti. After 3 years, the rhizobium population was altered; it showed a genetic structure based mainly on differences among plants, while the effects of soil and cultivar were not significant. PMID- 11055925 TI - Effect of temperature on carbon and electron flow and on the archaeal community in methanogenic rice field soil. AB - Temperature is an important factor controlling CH(4) production in anoxic rice soils. Soil slurries, prepared from Italian rice field soil, were incubated anaerobically in the dark at six temperatures of between 10 to 37 degrees C or in a temperature gradient block covering the same temperature range at intervals of 1 degrees C. Methane production reached quasi-steady state after 60 to 90 days. Steady-state CH(4) production rates increased with temperature, with an apparent activation energy of 61 kJ mol(-1). Steady-state partial pressures of the methanogenic precursor H(2) also increased with increasing temperature from <0.5 to 3.5 Pa, so that the Gibbs free energy change of H(2) plus CO(2)-dependent methanogenesis was kept at -20 to -25 kJ mol of CH(4)(-1) over the whole temperature range. Steady-state concentrations of the methanogenic precursor acetate, on the other hand, increased with decreasing temperature from <5 to 50 microM. Simultaneously, the relative contribution of H(2) as methanogenic precursor decreased, as determined by the conversion of radioactive bicarbonate to (14)CH(4), so that the carbon and electron flow to CH(4) was increasingly dominated by acetate, indicating that psychrotolerant homoacetogenesis was important. The relative composition of the archaeal community was determined by terminal restriction fragment length polymorphism (T-RFLP) analysis of the 16S rRNA genes (16S rDNA). T-RFLP analysis differentiated the archaeal Methanobacteriaceae, Methanomicrobiaceae, Methanosaetaceae, Methanosarcinaceae, and Rice clusters I, III, IV, V, and VI, which were all present in the rice field soil incubated at different temperatures. The 16S rRNA genes of Rice cluster I and Methanosaetaceae were the most frequent methanogenic groups. The relative abundance of Rice cluster I decreased with temperature. The substrates used by this microbial cluster, and thus its function in the microbial community, are unknown. The relative abundance of acetoclastic methanogens, on the other hand, was consistent with their physiology and the acetate concentrations observed at the different temperatures, i.e., the high-acetate-requiring Methanosarcinaceae decreased and the more modest Methanosaetaceae increased with increasing temperature. Our results demonstrate that temperature not only affected the activity but also changed the structure and the function (carbon and electron flow) of a complex methanogenic system. PMID- 11055926 TI - Engineering increased stability in the antimicrobial peptide pediocin PA-1. AB - Pediocin PA-1 is a food grade antimicrobial peptide that has been used as a food preservative. Upon storage at 4 degrees C or room temperature, pediocin PA-1 looses activity, and there is a concomitant 16-Da increase in the molecular mass. It is shown that the loss of activity follows first-order kinetics and that the instability can be prevented by replacing the single methionine residue (Met31) in pediocin PA-1. Replacing Met by Ala, Ile, or Leu protected the peptide from oxidation and had only minor effects on bacteriocin activity (for most indicator strains 100% activity was maintained). Replacement of Met by Asp was highly deleterious for bacteriocin activity. PMID- 11055927 TI - Molecular characterization of bacterial populations in petroleum-contaminated groundwater discharged from underground crude oil storage cavities. AB - Petroleum-contaminated groundwater discharged from underground crude oil storage cavities (cavity groundwater) harbored more than 10(6) microorganisms ml(-1), a density 100 times higher than the densities in groundwater around the cavities (control groundwater). To characterize bacterial populations growing in the cavity groundwater, 46 PCR-amplified almost full-length 16S ribosomal DNA (rDNA) fragments were cloned and sequenced, and 28 different sequences were obtained. All of the sequences were affiliated with the Proteobacteria; 25 sequences (43 clones) were affiliated with the epsilon subclass, 2 were affiliated with the beta subclass, and 1 was affiliated with the delta subclass. Two major clusters (designated clusters 1 and 2) were found for the epsilon subclass proteobacterial clones; cluster 1 (25 clones) was most closely related to Thiomicrospira denitrificans (88% identical in nucleotide sequence), while cluster 2 (11 clones) was closely related to Arcobacter spp. Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified partial 16S rDNA fragments showed that one band was detected most strongly in cavity groundwater profiles independent of storage oil type and season. The sequence of this major band was identical to the sequences of most of the cluster 1 clones. Fluorescence in situ hybridization (FISH) indicated that the cluster 1 population accounted for 12 to 24% of the total bacterial population. This phylotype was not detected in the control groundwater by DGGE and FISH analyses. These results indicate that the novel members of the epsilon subclass of the Proteobacteria grow as major populations in the petroleum contaminated cavity groundwater. PMID- 11055928 TI - xylP promoter-based expression system and its use for antisense downregulation of the Penicillium chrysogenum nitrogen regulator NRE. AB - A highly inducible fungal promoter derived from the Penicillium chrysogenum endoxylanase (xylP) gene is described. Northern analysis and the use of a beta glucuronidase (uidA) reporter gene strategy showed that xylP expression is transcriptionally regulated. Xylan and xylose are efficient inducers, whereas glucose strongly represses the promoter activity. Comparison of the same expression construct as a single copy at the niaD locus in P. chrysogenum and at the argB locus in Aspergillus nidulans demonstrated that the xylP promoter is regulated similarly in these two species but that the level of expression is about 80 times higher in the Aspergillus species. The xylP promoter was found to be 65-fold more efficient than the isopenicillin-N-synthetase (pcbC) promoter in Penicillium and 23-fold more efficient than the nitrate reductase (niaD) promoter in Aspergillus under induced conditions. Furthermore, the xylP promoter was used for controllable antisense RNA synthesis of the nre-encoded putative major nitrogen regulator of P. chrysogenum. This approach led to inducible downregulation of the steady-state mRNA level of nre and consequently to transcriptional repression of the genes responsible for nitrate assimilation. In addition, transcription of nreB, which encodes a negative-acting nitrogen regulatory GATA factor of Penicillium, was found to be subject to regulation by NRE. Our data are the first direct evidence that nre indeed encodes an activator in the nitrogen regulatory circuit in Penicillium and indicate that cross regulation of the controlling factors occurs. PMID- 11055929 TI - Advances in development of a genetic system for Thermoanaerobacterium spp.: expression of genes encoding hydrolytic enzymes, development of a second shuttle vector, and integration of genes into the chromosome. AB - Despite recent success in transforming various thermophilic gram-type-positive anaerobes with plasmid DNA, use of shuttle vectors for the expression of genes other than antibiotic resistance markers has not previously been described. We constructed new vectors in order to express heterologous hydrolytic enzymes in our model system, Thermoanaerobacterium saccharolyticum JW/SL-YS485. Transformed Thermoanaerobacterium expressed active enzyme, indicating that this system may function as an alternate expression host, especially for genes with a thermophilic origin. To develop further the genetic system for T. saccharolyticum JW/SL-YS485, two improved Escherichia coli-Thermoanaerobacterium shuttle vectors, pRKM1 and pRUKM, were constructed. Furthermore, the kanamycin resistance cassette alone and the kanamycin resistance cassette plus the cellobiohydrolase gene (cbhA) from Clostridium thermocellum JW20 were integrated into the xylanase gene (xynA) region of the Thermoanaerobacterium chromosome via homologous recombination using pUC-based suicide vectors pUXK and pUXKC. PMID- 11055930 TI - Integrative food-grade expression system based on the lactose regulon of Lactobacillus casei. AB - The lactose operon from Lactobacillus casei is regulated by very tight glucose repression and substrate induction mechanisms, which made it a tempting candidate system for the expression of foreign genes or metabolic engineering. An integrative vector was constructed, allowing stable gene insertion in the chromosomal lactose operon of L. casei. This vector was based on the nonreplicative plasmid pRV300 and contained two DNA fragments corresponding to the 3' end of lacG and the complete lacF gene. Four unique restriction sites were created, as well as a ribosome binding site that would allow the cloning and expression of new genes between these two fragments. Then, integration of the cloned genes into the lactose operon of L. casei could be achieved via homologous recombination in a process that involved two selection steps, which yielded highly stable food-grade mutants. This procedure has been successfully used for the expression of the E. coli gusA gene and the L. lactis ilvBN genes in L. casei. Following the same expression pattern as that for the lactose genes, beta glucuronidase activity and diacetyl production were repressed by glucose and induced by lactose. This integrative vector represents a useful tool for strain improvement in L. casei that could be applied to engineering fermentation processes or used for expression of genes for clinical and veterinary uses. PMID- 11055931 TI - Marine planktonic archaea take up amino acids. AB - Archaea are traditionally thought of as "extremophiles," but recent studies have shown that marine planktonic Archaea make up a surprisingly large percentage of ocean midwater microbial communities, up to 60% of the total prokaryotes. However, the basic physiology and contribution of Archaea to community microbial activity remain unknown. We have studied Archaea from 200-m depths of the northwest Mediterranean Sea and the Pacific Ocean near California, measuring the archaeal activity under simulated natural conditions (8 to 17 degrees C, dark and aerobic [corrected]) by means of a method called substrate tracking autoradiography fluorescence in situ hybridization (STARFISH) that simultaneously detects specific cell types by 16S rRNA probe binding and activity by microautoradiography. In the 200-m-deep Mediterranean and Pacific samples, cells binding the archaeal probes made up about 43 and 14% of the total countable cells, respectively. Our results showed that the Archaea are active in the uptake of dissolved amino acids from natural concentrations (nanomolar) with about 60% of the individuals in the archaeal communities showing measurable uptake. Bacteria showed a similar proportion of active cells. We concluded that a portion of these Archaea is heterotrophic and also appears to coexist successfully with Bacteria in the same water. PMID- 11055932 TI - Application of siderotyping for characterization of Pseudomonas tolaasii and "Pseudomonas reactans" isolates associated with brown blotch disease of cultivated mushrooms. AB - Pyoverdine isoelectric focusing analysis and pyoverdine-mediated iron uptake were used as siderotyping methods to analyze a collection of 57 northern and central European isolates of P. tolaasii and "P. reactans." The bacteria, isolated from cultivated Agaricus bisporus or Pleurotus ostreatus mushroom sporophores presenting brown blotch disease symptoms, were identified according to the white line test (W. C. Wong and T. F. Preece, J. Appl. Bacteriol. 47:401-407, 1979) and their pathogenicity towards A. bisporus and were grouped into siderovars according to the type of pyoverdine they produced. Seventeen P. tolaasii isolates were recognized, which divided into two siderovars, with the first one containing reference strains and isolates of various geographical origins while the second one contained Finnish isolates exclusively. The 40 "P. reactans" isolates divided into eight siderovars. Pyoverdine isoelectric focusing profiles and cross-uptake studies demonstrated an identity for some "P. reactans" isolates, with reference strains belonging to the P. fluorescens biovars II, III, or V. Thus, the easy and rapid methods of siderotyping proved to be reliable by supporting and strengthening previous taxonomical data. Moreover, two potentially novel pyoverdines characterizing one P. tolaasii siderovar and one "P. reactans" siderovar were found. PMID- 11055933 TI - Characterization and chromosomal mapping of antimicrobial resistance genes in Salmonella enterica serotype typhimurium. AB - Two hundred and twenty-six Salmonella enterica serotype Typhimurium isolates were examined for the presence of integron-associated gene cassettes. All but two of the non-DT104 isolates, together with DT104 isolates, contained gene cassettes. Amplicons of 1.5 kbp each were found in two non-DT104 isolates, encoding a dhfrI gene (trimethoprim resistance) linked to an aadA gene (streptomycin and spectinomycin resistance), by site-specific recombination. DT104 isolates of resistance (R) type ACSSuT possessed the recently described 1.0- and 1.2-kbp gene cassettes. Macrorestriction analysis with XbaI and DNA probing mapped ant(3")-1a, bla(PSE-1), and dhfrI genes to large multiresistant gene clusters in a DT170a isolate and a DT193 isolate. In contrast, all DT104 isolates (R-type ACSSuT) possessed a conserved 10-kbp Xba1 DNA fragment. Our study highlights the occurrence of integrons (and antimicrobial resistance determinants) among serotype Typhimurium isolates other than DT104. Larger and previously unrecognized multiresistance gene clusters were identified in these isolates by DNA probing. PMID- 11055934 TI - Fate of selenate and selenite metabolized by Rhodobacter sphaeroides. AB - Cultures of a purple nonsulfur bacterium, Rhodobacter sphaeroides, amended with approximately 1 or approximately 100 ppm selenate or selenite, were grown phototrophically to stationary phase. Analyses of culture headspace, separated cells, and filtered culture supernatant were carried out using gas chromatography, X-ray absorption spectroscopy, and inductively coupled plasma spectroscopy-mass spectrometry, respectively. While selenium-amended cultures showed much higher amounts of SeO(3)(2-) bioconversion than did analogous selenate experiments (94% uptake for SeO(3)(2-) as compared to 9.6% for SeO(4)(2 )-amended cultures from 100-ppm solutions), the chemical forms of selenium in the microbial cells were not very different except at exposure to high concentrations of selenite. Volatilization accounted for only a very small portion of the accumulated selenium; most was present in organic forms and the red elemental form. PMID- 11055935 TI - Exogenous isolation of antibiotic resistance plasmids from piggery manure slurries reveals a high prevalence and diversity of IncQ-like plasmids. AB - Antibiotic resistance plasmids were exogenously isolated in biparental matings with piggery manure bacteria as plasmid donors in Escherichia coli CV601 and Pseudomonas putida UWC1 recipients. Surprisingly, IncQ-like plasmids were detected by dot blot hybridization with an IncQ oriV probe in several P. putida UWC1 transconjugants. The capture of IncQ-like plasmids in biparental matings indicates not only their high prevalence in manure slurries but also the presence of efficiently mobilizing plasmids. In order to elucidate unusual hybridization data (weak or no hybridization with IncQ repB or IncQ oriT probes) four IncQ-like plasmids (pIE1107, pIE1115, pIE1120, and pIE1130), each representing a different EcoRV restriction pattern, were selected for a more thorough plasmid characterization after transfer into E. coli K-12 strain DH5alpha by transformation. The characterization of the IncQ-like plasmids revealed an astonishingly high diversity with regard to phenotypic and genotypic properties. Four different multiple antibiotic resistance patterns were found to be conferred by the IncQ-like plasmids. The plasmids could be mobilized by the RP4 derivative pTH10 into Acinetobacter sp., Ralstonia eutropha, Agrobacterium tumefaciens, and P. putida, but they showed diverse patterns of stability under nonselective growth conditions in different host backgrounds. Incompatibility testing and PCR analysis clearly revealed at least two different types of IncQ-like plasmids. PCR amplification of total DNA extracted directly from different manure samples and other environments indicated the prevalence of both types of IncQ plasmids in manure, sewage, and farm soil. These findings suggest that IncQ plasmids play an important role in disseminating antibiotic resistance genes. PMID- 11055936 TI - Variation in tolerance and virulence in the chestnut blight fungus-hypovirus interaction. AB - Chestnut blight, caused by the fungus Cryphonectria parasitica, has been effectively controlled with double-stranded RNA hypoviruses in Europe for over 40 years. The marked reduction in the virulence of C. parasitica by hypoviruses is a phenomenon known as hypovirulence. This virus-fungus pathosystem has become a model system for the study of biological control of fungi with viruses. We studied variation in tolerance to hypoviruses in fungal hosts and variation in virulence among virus isolates from a local population in Italy. Tolerance is defined as the relative fitness of a fungal individual when infected with hypoviruses (compared to being uninfected); virulence is defined for each hypovirus as the reduction in fitness of fungal hosts relative to virus-free hosts. Six hypovirus-infected isolates of C. parasitica were sampled from the population, and each hypovirus was transferred into six hypovirus-free recipient isolates. The resulting 36 hypovirus-fungus combinations were used to estimate genetic variation in tolerance to hypoviruses, in hypovirus virulence, and in virus-fungus interactions. Four phenotypes were evaluated for each virus-fungus combination to estimate relative fitness: (i) sporulation, i.e., the number of asexual spores (conidia) produced; (ii) canker area on field-inoculated chestnut trees, (iii) vertical transmission of hypoviruses into conidia, and (iv) conidial germination. Two-way analysis of variance (ANOVA) revealed significant interactions (P < 0.001) between viruses and fungal isolates for sporulation and canker area but not for conidial germination or transmission. One-way ANOVA among hypoviruses (within each fungal isolate) and among fungal isolates (within each hypovirus) revealed significant genetic variation (P < 0.01) in hypovirus virulence and fungal tolerance within several fungal isolates, and hypoviruses, respectively. These interactions and the significant genetic variation in several fitness characters indicate the potential for future evolution of these characters. However, biological control is unlikely to break down due to evolution of tolerance to hypoviruses in the fungus because the magnitudes of tolerance and interactions were relatively small. PMID- 11055937 TI - Evidence of substantial carbon isotope fractionation among substrate, inorganic carbon, and biomass during aerobic mineralization of 1, 2-dichloroethane by Xanthobacter autotrophicus. AB - Carbon isotope fractionation during aerobic mineralization of 1, 2-dichloroethane (1,2-DCA) by Xanthobacter autotrophicus GJ10 was investigated. A strong enrichment of (13)C in residual 1,2-DCA was observed, with a mean fractionation factor alpha +/- standard deviation of 0.968 +/- 0.0013 to 0.973 +/- 0.0015. In addition, a large carbon isotope fractionation between biomass and inorganic carbon occurred. A mechanistic model that links the fractionation factor alpha to the rate constants of the first catabolic enzyme was developed. Based on the model, it was concluded that the strong enrichment of (13)C in 1,2-DCA arises because the first irreversible step of the initial enzymatic transformation of 1,2-DCA consists of an S(N)2 nucleophilic substitution. S(N)2 reactions are accompanied by a large kinetic isotope effect. The substantial carbon isotope fractionation between biomass and inorganic carbon could be explained by the kinetic isotope effect associated with the initial 1,2-DCA transformation and by the metabolic pathway of 1,2-DCA degradation. Carbon isotope fractionation during 1,2-DCA mineralization leads to 1,2-DCA, inorganic carbon, and biomass with characteristic carbon isotope compositions, which may be used to trace the process in contaminated environments. PMID- 11055938 TI - Purification and characterization of Streptomyces griseus catechol O methyltransferase. AB - A soluble (100,000 x g supernatant) methyltransferase catalyzing the transfer of the methyl group of S-adenosyl-L-methionine to catechols was present in cell extracts of Streptomyces griseus. A simple, general, and rapid catechol-based assay method was devised for enzyme purification and characterization. The enzyme was purified 141-fold by precipitation with ammonium sulfate and successive chromatography over columns of DEAE-cellulose, DEAE-Sepharose, and Sephacryl S 200. The purified cytoplasmic enzyme required 10 mM magnesium for maximal activity and was catalytically optimal at pH 7. 5 and 35 degrees C. The methyltransferase had an apparent molecular mass of 36 kDa for both the native and denatured protein, with a pI of 4.4. Novel N-terminal and internal amino acid sequences were determined as DFVLDNEGNPLENNGGYXYI and RPDFXLEPPYTGPXKARIIRYFY, respectively. For this enzyme, the K(m) for 6,7-dihydroxycoumarin was 500 +/- 21.5 microM, and that for S-adenosyl-L-methionine was 600 +/- 32.5 microM. Catechol, caffeic acid, and 4-nitrocatechol were methyltransferase substrates. Homocysteine was a competitive inhibitor of S-adenosyl-L-methionine, with a K(i) of 224 +/- 20.6 microM. Sinefungin and S-adenosylhomocysteine inhibited methylation, and the enzyme was inactivated by Hg(2+), p-chloromercuribenzoic acid, and N-ethylmaleimide. PMID- 11055939 TI - Screening of genes involved in isooctane tolerance in Saccharomyces cerevisiae by using mRNA differential display. AB - A Saccharomyces cerevisiae strain, KK-211, isolated by the long-term bioprocess of stereoselective reduction in isooctane, showed extremely high tolerance to the solvent, which is toxic to yeast cells, but, in comparison with its wild-type parent, DY-1, showed low tolerance to hydrophilic organic solvents, such as dimethyl sulfoxide and ethanol. In order to detect the isooctane tolerance associated genes, mRNA differential display (DD) was employed using mRNAs isolated from strains DY-1 and KK-211 cultivated without isooctane, and from strain KK-211 cultivated with isooctane. Thirty genes were identified as being differentially expressed in these three types of cells and were classified into three groups according to their expression patterns. These patterns were further confirmed and quantified by Northern blot analysis. On the DD fingerprints, the expression of 14 genes, including MUQ1, PRY2, HAC1, AGT1, GAC1, and ICT1 (YLR099c) was induced, while the expression of the remaining 16 genes, including JEN1, PRY1, PRY3, and KRE1, was decreased, in strain KK-211 cultivated with isooctane. The genes represented by HAC1, PRY1, and ICT1 have been reported to be associated with cell stress, and AGT1 and GAC1 have been reported to be involved in the uptake of trehalose and the production of glycogen, respectively. MUQ1 and KRE1, encoding proteins associated with cell surface maintenance, were also detected. Based on these results, we concluded that alteration of expression levels of multiple genes, not of a single gene, might be the critical determinant for isooctane tolerance in strain KK-211. PMID- 11055940 TI - Use of hydrostatic pressure for inactivation of microbial contaminants in cheese. AB - The objective of this study was to determine the effect of high pressure (HP) on the inactivation of microbial contaminants in Cheddar cheese (Escherichia coli K 12, Staphylococcus aureus ATCC 6538, and Penicillium roqueforti IMI 297987). Initially, cheese slurries inoculated with E. coli, S. aureus, and P. roqueforti were used as a convenient means to define the effects of a range of pressures and temperatures on the viability of these microorganisms. Cheese slurries were subjected to pressures of 50 to 800 MPa for 20 min at temperatures of 10, 20, and 30 degrees C. At 400 MPa, the viability of P. roqueforti in cheese slurry decreased by >2-log-unit cycles at 10 degrees C and by 6-log-unit cycles at temperatures of 20 and 30 degrees C. S. aureus and E. coli were not detected after HP treatments in cheese slurry of >600 MPa at 20 degrees C and >400 MPa at 30 degrees C, respectively. In addition to cell death, the presence of sublethally injured cells in HP-treated slurries was demonstrated by differential plating using nonselective agar incorporating salt or glucose. Kinetic experiments of HP inactivation demonstrated that increasing the pressure from 300 to 400 MPa resulted in a higher degree of inactivation than increasing the pressurization time from 0 to 60 min, indicating a greater antimicrobial impact of pressure. Selected conditions were subsequently tested on Cheddar cheese by adding the isolates to cheese milk and pressure treating the resultant cheeses at 100 to 500 MPa for 20 min at 20 degrees C. The relative sensitivities of the isolates to HP in Cheddar cheese were similar to those observed in the cheese slurry, i.e., P. roqueforti was more sensitive than E. coli, which was more sensitive than S. aureus. The organisms were more sensitive to pressure in cheese than slurry, especially with E. coli. On comparison of the sensitivities of the microorganisms in a pH 5.3 phosphate buffer, cheese slurry, and Cheddar cheese, greatest sensitivity to HP was shown in the pH 5.3 phosphate buffer by S. aureus and P. roqueforti while greatest sensitivity to HP by E. coli was exhibited in Cheddar cheese. Therefore, the medium in which the microorganisms are treated is an important determinant of the level of inactivation observed. PMID- 11055941 TI - Complete nucleotide sequence of ubiquitous plasmid pEA29 from Erwinia amylovora strain Ea88: gene organization and intraspecies variation. AB - The complete sequence of plasmid pEA29 from Erwinia amylovora strain Ea88 consists of 28,185 bp with a 50.2% G+C content. As deletions and insertions were detected in other derivatives of pEA29, its size actually varied from 27.6 to 34.9 kb. Thirteen open reading frames that encoded predicted proteins with similarities to known proteins from other bacteria were identified along with two open reading frames related to hypothetical proteins found in GenBank and six open reading frames with no similarities to existing GenBank entries. Predicted products of open reading frames with similarity to the thiamine biosynthetic genes thiO, thiG, and thiF; a betT gene coding for choline transport; an msrA gene for the enzyme methionine sulfoxide reductase; a putative methyl-accepting chemotaxis gene; an aldehyde dehydrogenase gene; an hns DNA binding gene; a LysR type transcriptional regulator; and parA and parB partitioning genes were identified. A putative iteron-containing theta-type origin of replication with an AT-rich region and a gene for a RepA protein was identified. PstI and KpnI restriction patterns for pEA29 isolated from tree fruit strains of E. amylovora were homogenous and different from those for pEA29 isolated from Rubus (raspberry) strains. All Rubus derivatives of pEA29 contained a point mutation that eliminated a PstI site and a 1,264-bp region that replaced 1, 890 bp of sequence found in pEA29 from strain Ea88. This change eliminated a second PstI site and increased the length of a KpnI fragment. An insertion sequence, ISEam1, was detected in one Rubus strain, and transposon Tn5393 was detected in three apple strains in two separate locations on the plasmid. Plasmid-cured strains exhibited reduced virulence and modified colony morphology on minimal medium without thiamine, indicating that some of the genes in pEA29 play a role in the physiology or metabolism of E. amylovora. PMID- 11055942 TI - Occurrence of antimicrobial resistance in fish-pathogenic and environmental bacteria associated with four danish rainbow trout farms. AB - Surveillance of bacterial susceptibility to five antimicrobial agents was performed during a 1-year period in and around four freshwater fish farms situated along a stream in western Denmark. Besides assessing the levels of antibiotic resistance among the culturable fraction of microorganisms in fish, water, and sediment samples, two major fish pathogens (88 Flavobacterium psychrophilum isolates and 134 Yersinia ruckeri isolates) and 313 motile Aeromonas isolates, representing a group of ubiquitous aquatic bacteria, were isolated from the same samples. MICs were obtained applying a standardized agar dilution method. A markedly decreased susceptibility of F. psychrophilum isolates to most antimicrobial agents presently available for use in Danish aquaculture was detected, while the collected Y. ruckeri isolates remained largely sensitive to all therapeutic substances. Comparing the inlet and outlet samples, the increase of the antibiotic-resistant proportions observed among the culturable microflora was more pronounced and statistically significant among the motile aeromonads. High levels of individual and multiple antimicrobial resistances were demonstrated within the collected flavobacteria and aeromonads, thus indicating a substantial impact of fish farming on several groups of bacteria associated with aquacultural environments. PMID- 11055943 TI - Viral impacts on total abundance and clonal composition of the harmful bloom forming phytoplankton Heterosigma akashiwo. AB - Recent observations that viruses are very abundant and biologically active components in marine ecosystems suggest that they probably influence various biogeochemical and ecological processes. In this study, the population dynamics of the harmful bloom-forming phytoplankton Heterosigma akashiwo (Raphidophyceae) and the infectious H. akashiwo viruses (HaV) were monitored in Hiroshima Bay, Japan, from May to July 1998. Concurrently, a number of H. akashiwo and HaV clones were isolated, and their virus susceptibilities and host ranges were determined through laboratory cross-reactivity tests. A sudden decrease in cell density of H. akashiwo was accompanied by a drastic increase in the abundance of HaV, suggesting that viruses contributed greatly to the disintegration of the H. akashiwo bloom as mortality agents. Despite the large quantity of infectious HaV, however, a significant proportion of H. akashiwo cells survived after the bloom disintegration. The viral susceptibility of H. akashiwo isolates demonstrated that the majority of these surviving cells were resistant to most of the HaV clones, whereas resistant cells were a minor component during the bloom period. Moreover, these resistant cells were displaced by susceptible cells, presumably due to viral infection. These results demonstrated that the properties of dominant cells within the H. akashiwo population change during the period when a bloom is terminated by viral infection, suggesting that viruses also play an important role in determining the clonal composition and maintaining the clonal diversity of H. akashiwo populations. Therefore, our data indicate that viral infection influences the total abundance and the clonal composition of one host algal species, suggesting that viruses are an important component in quantitatively and qualitatively controlling phytoplankton populations in natural marine environments. PMID- 11055944 TI - Habituation of Salmonella spp. at reduced water activity and its effect on heat tolerance. AB - The effect of habituation at reduced water activity (a(w)) on heat tolerance of Salmonella spp. was investigated. Stationary-phase cells were exposed to a(w) 0.95 in broths containing glucose-fructose, sodium chloride, or glycerol at 21 degrees C for up to a week prior to heat challenge at 54 degrees C. In addition, the effects of different a(w)s and heat challenge temperatures were investigated. Habituation at a(w) 0.95 resulted in increased heat tolerance at 54 degrees C with all solutes tested. The extent of the increase and the optimal habituation time depended on the solute used. Exposure to broths containing glucose-fructose (a(w) 0.95) for 12 h resulted in maximal heat tolerance, with more than a fourfold increase in D(54) values. Cells held for more than 72 h in these conditions, however, became as heat sensitive as nonhabituated populations. Habituation in the presence of sodium chloride or glycerol gave rise to less pronounced but still significant increases in heat tolerance at 54 degrees C, and a shorter incubation time was required to maximize tolerance. The increase in heat tolerance following habituation in broths containing glucose-fructose (a(w) 0.95) was RpoS independent. The presence of chloramphenicol or rifampin during habituation and inactivation did not affect the extent of heat tolerance achieved, suggesting that de novo protein synthesis was probably not necessary. These data highlight the importance of cell prehistory prior to heat inactivation and may have implications for food manufacturers using low-a(w) ingredients. PMID- 11055945 TI - Persistent colonization of sheep by Escherichia coli O157:H7 and other E. coli pathotypes. AB - Shiga toxin-producing Escherichia coli (STEC) is an important cause of food-borne illness in humans. Ruminants appear to be more frequently colonized by STEC than are other animals, but the reason(s) for this is unknown. We compared the frequency, magnitude, duration, and transmissibility of colonization of sheep by E. coli O157:H7 to that by other pathotypes of E. coli. Young adult sheep were simultaneously inoculated with a cocktail consisting of two strains of E. coli O157:H7, two strains of enterotoxigenic E. coli (ETEC), and one strain of enteropathogenic E. coli. Both STEC strains and ETEC 2041 were given at either 10(7) or 10(10) CFU/strain/animal. The other strains were given only at 10(10) CFU/strain. We found no consistent differences among pathotypes in the frequency, magnitude, and transmissibility of colonization. However, the STEC strains tended to persist to 2 weeks and 2 months postinoculation more frequently than did the other pathotypes. The tendency for persistence of the STEC strains was apparent following an inoculation dose of either 10(7) or 10(10) CFU. One of the ETEC strains also persisted when inoculated at 10(10) CFU. However, in contrast to the STEC strains, it did not persist when inoculated at 10(7) CFU. These results support the hypothesis that STEC is better adapted to persist in the alimentary tracts of sheep than are other pathotypes of E. coli. PMID- 11055946 TI - Monitoring bacterial transport by stable isotope enrichment of cells. AB - Understanding the transport and behavior of bacteria in the environment has broad implications in diverse areas, ranging from agriculture to groundwater quality, risk assessment, and bioremediation. The ability to reliably track and enumerate specific bacterial populations in the context of native communities and environments is key to developing this understanding. We report a novel bacterial tracking approach, based on altering the stable carbon isotope value (delta(13)C) of bacterial cells, which provides specific and sensitive detection and quantification of those cells in environmental samples. This approach was applied to the study of bacterial transport in saturated porous media. The transport of introduced organisms was indicated by mass spectrometric analysis of groundwater samples, where the presence of (13)C-enriched bacteria resulted in increased delta(13)C values of the samples, allowing specific and sensitive detection and enumeration of the bacteria of interest. We demonstrate the ability to produce highly (13)C-enriched bacteria, present data indicating that results obtained with this approach accurately represent intact introduced bacteria, and include field data on the use of this stable isotope approach to monitor in situ bacterial transport. This detection strategy allows sensitive detection of an introduced, unmodified bacterial strain in the presence of the indigenous bacterial community, including itself in its unenriched form. PMID- 11055947 TI - Introduction of an N-glycosylation site increases secretion of heterologous proteins in yeasts. AB - Saccharomyces cerevisiae is often used to produce heterologous proteins that are preferentially secreted to increase economic feasibility. We used N-glycosylation as a tool to enhance protein secretion. Secretion of cutinase, a lipase, and llama V(HH) antibody fragments by S. cerevisiae or Pichia pastoris improved following the introduction of an N-glycosylation site. When we introduced an N glycosylation consensus sequence in the N-terminal region of a hydrophobic cutinase, secretion increased fivefold. If an N-glycosylation site was introduced in the C-terminal region, however, secretion increased only 1.8-fold. These results indicate that the use of N glycosylation can significantly enhance heterologous protein secretion. PMID- 11055948 TI - PCR bias in ecological analysis: a case study for quantitative Taq nuclease assays in analyses of microbial communities. AB - Succession of ecotypes, physiologically diverse strains with negligible rRNA sequence divergence, may explain the dominance of small, red-pigmented (phycoerythrin-rich) cyanobacteria in the autotrophic picoplankton of deep lakes (C. Postius and A. Ernst, Arch. Microbiol. 172:69-75, 1999). In order to test this hypothesis, it is necessary to determine the abundance of specific ecotypes or genotypes in a mixed background of phylogenetically similar organisms. In this study, we examined the performance of Taq nuclease assays (TNAs), PCR-based assays in which the amount of an amplicon is monitored by hydrolysis of a labeled oligonucleotide (TaqMan probe) when hybridized to the amplicon. High accuracy and a 7-order detection range made the real-time TNA superior to the corresponding end point technique. However, in samples containing mixtures of homologous target sequences, quantification can be biased due to limited specificity of PCR primers and probe oligonucleotides and due to accumulation of amplicons that are not detected by the TaqMan probe. A decrease in reaction efficiency, which can be recognized by direct monitoring of amplification, provides experimental evidence for the presence of such a problem and emphasizes the need for real-time technology in quantitative PCR. Use of specific primers and probes and control of amplification efficiency allow correct quantification of target DNA in the presence of an up to 10(4)-fold excess of phylogenetically similar DNA and of an up to 10(7)-fold excess of dissimilar DNA. PMID- 11055949 TI - Population structure of rat-derived Pneumocystis carinii in Danish wild rats. AB - The rat model of Pneumocystis carinii pneumonia is frequently used to study human P. carinii infection, but there are many differences between the rat and human infections. We studied naturally acquired P. carinii in wild rats to examine the relevance of the rat model for human infection. P. carinii DNA was detected in 47 of 51 wild rats and in 10 of 12 nonimmunosuppressed laboratory rats. Evidence for three novel formae speciales of rat-derived P. carinii was found, and these were provisionally named Pneumocystis carinii f. sp. rattus-secundi, Pneumocystis carinii f. sp. rattus-tertii, and Pneumocystis carinii f. sp. rattus-quarti. Our data suggest that low-level carriage of P. carinii in wild rats and nonimmunosuppressed laboratory rats is common and that wild rats are frequently coinfected with more than one forma specialis of P. carinii. We also examined the diversity in the internally transcribed spacer (ITS) regions of the nuclear rRNA operon of Pneumocystis carinii f. sp. carinii by using samples from wild rats and laboratory rats and spore trap samples. We report a lack of variation in the ITS1 and ITS2 regions that is consistent with an evolutionary bottleneck in the P. carinii f. sp. carinii population. This study shows that human- and rat-derived P. carinii organisms are very different, not only in genetic composition but also in population structure and natural history. PMID- 11055950 TI - Comparison of acid mine drainage microbial communities in physically and geochemically distinct ecosystems. AB - This study presents population analyses of microbial communities inhabiting a site of extreme acid mine drainage (AMD) production. The site is the inactive underground Richmond mine at Iron Mountain, Calif., where the weathering of a massive sulfide ore body (mostly pyrite) produces solutions with pHs of approximately 0.5 to approximately 1.0. Here we used a suite of oligonucleotide probes, designed from molecular data recently acquired from the site, to analyze a number of microbial environments by fluorescent in situ hybridization. Microbial-community analyses were correlated with geochemical and mineralogical data from those environments. The environments investigated were within the ore body and thus at the site of pyrite dissolution, as opposed to environments that occur downstream of the dissolution. Few organism types, as defined by the specificities of the oligonucleotide probes, dominated the microbial communities. The majority of the dominant organisms detected were newly discovered or organisms only recently associated with acid-leaching environments. "Ferroplasma" spp. were detected in many of the communities and were particularly dominant in environments of lowest pH and highest ionic strength. Leptospirillum spp. were also detected in many slime and pyrite-dominated environments. In samples of an unusual subaerial slime, a new uncultured Leptospirillum sp. dominated. Sulfobacillus spp. were detected as a prominent inhabitant in warmer ( approximately 43 degrees C) environments. The information gathered here is critical for determining organisms important to AMD production at Iron Mountain and for directing future studies of this process. The findings presented here also have relevance to the microbiology of industrial bioleaching and to the understanding of geochemical iron and sulfur cycles. PMID- 11055951 TI - A novel selenite- and tellurite-inducible gene in Escherichia coli. AB - Selenium is both an essential and a toxic trace element, and the range of concentrations between the two is extremely narrow. Although tellurium is not essential and is only rarely found in the environment, it is considered to be extremely toxic. Several hypotheses have been proposed to account for the toxic effects of selenite and tellurite. However, these potential mechanisms have yet to be fully substantiated. Through screening of an Escherichia coli luxAB transcriptional gene fusion library, we identified a clone whose luminescence increased in the presence of increasing concentrations of sodium selenite or sodium tellurite. Cloning and sequencing of the luxAB junction revealed that the fusion had occurred in a previously uncharacterized open reading frame, termed o393 or yhfC, which we have now designated gutS, for gene up-regulated by tellurite and selenite. Transcription from gutS in the presence of selenite or tellurite was confirmed by RNA dot blot analysis. In vivo expression of the GutS polypeptide, using the pET expression system, revealed a polypeptide of approximately 43 kDa, in good agreement with its predicted molecular mass. Although the function of GutS remains to be elucidated, homology searches as well as protein motif and secondary-structure analyses have provided clues which may implicate GutS in transport in response to selenite and tellurite. PMID- 11055952 TI - Growth limits of Listeria monocytogenes as a function of temperature, pH, NaCl, and lactic acid. AB - Models describing the limits of growth of pathogens under multiple constraints will aid management of the safety of foods which are sporadically contaminated with pathogens and for which subsequent growth of the pathogen would significantly increase the risk of food-borne illness. We modeled the effects of temperature, water activity, pH, and lactic acid levels on the growth of two strains of Listeria monocytogenes in tryptone soya yeast extract broth. The results could be divided unambiguously into "growth is possible" or "growth is not possible" classes. We observed minor differences in growth characteristics of the two L. monocytogenes strains. The data follow a binomial probability distribution and may be modeled using logistic regression. The model used is derived from a growth rate model in a manner similar to that described in a previously published work (K. A. Presser, T. Ross, and D. A. Ratkowsky, Appl. Environ. Microbiol. 64:1773-1779, 1998). We used "nonlinear logistic regression" to estimate the model parameters and developed a relatively simple model that describes our experimental data well. The fitted equations also described well the growth limits of all strains of L. monocytogenes reported in the literature, except at temperatures beyond the limits of the experimental data used to develop the model (3 to 35 degrees C). The models developed will improve the rigor of microbial food safety risk assessment and provide quantitative data in a concise form for the development of safer food products and processes. PMID- 11055953 TI - Ectomycorrhizal specificity patterns in a mixed Pinus contorta and Picea engelmannii forest in Yellowstone National Park. AB - We used molecular genetic methods to test two hypotheses, (i) that host plant specificity among ectomycorrhizal fungi would be common in a closed-canopy, mixed Pinus contorta-Picea engelmannii forest in Yellowstone National Park and (ii) that specificity would be more common in the early successional tree species, P. contorta, than in the invader, P. engelmannii. We identified 28 ectomycorrhizal fungal species collected from 27 soil cores. The proportion of P. engelmannii to P. contorta ectomycorrhizae was nearly equal (52 and 48%, respectively). Of the 28 fungal species, 18 composed greater than 95% of the fungal community. No species was associated exclusively with P. contorta, but four species, each found in only one core, and one species found in two cores were associated exclusively with P. engelmannii. These fungi composed less than 5% of the total ectomycorrhizae. Thus, neither hypothesis was supported, and hypothesized benefits of ectomycorrhizal specificity to both trees and fungi probably do not exist in this system. PMID- 11055954 TI - Genetic analysis of type E botulinum toxin-producing Clostridium butyricum strains. AB - Type E botulinum toxin (BoNT/E)-producing Clostridium butyricum strains isolated from botulism cases or soil specimens in Italy and China were analyzed by using nucleotide sequencing of the bont/E gene, random amplified polymorphic DNA (RAPD) assay, pulsed-field gel electrophoresis (PFGE), and Southern blot hybridization for the bont/E gene. Nucleotide sequences of the bont/E genes of 11 Chinese isolates and of the Italian strain BL 6340 were determined. The nucleotide sequences of the bont/E genes of 11 C. butyricum isolates from China were identical. The deduced amino acid sequence of BoNT/E from the Chinese isolates showed 95.0 and 96.9% identity with those of BoNT/E from C. butyricum BL 6340 and Clostridium botulinum type E, respectively. The BoNT/E-producing C. butyricum strains were divided into the following three clusters based on the results of RAPD assay, PFGE profiles of genomic DNA digested with SmaI or XhoI, and Southern blot hybridization: strains associated with infant botulism in Italy, strains associated with food-borne botulism in China, and isolates from soil specimens of the Weishan lake area in China. A DNA probe for the bont/E gene hybridized with the nondigested chromosomal DNA of all toxigenic strains tested, indicating chromosomal localization of the bont/E gene in C. butyricum. The present results suggest that BoNT/E-producing C. butyricum is clonally distributed over a vast area. PMID- 11055955 TI - Purification and properties of an enzyme capable of degrading the sheath of Sphaerotilus natans. AB - Microorganisms which can degrade and grow on the purified sheath of a sheathed bacterium Sphaerotilus natans were collected from soil and river water. Two bacterial strains were isolated from the soil and designated strains TB and TK. Both strains are rod shaped, negatively stained by gram staining, facultatively anaerobic, and formed ellipsoidal endospores. These characteristics suggested that the isolates belong to the genus Paenibacillus, according to Ash et al. (C. Ash, F. G. Priest, and M. D. Collins, Antonie Leeuwenhoek 64:253-260, 1993). Phylogenetic analysis based on the 16S rDNA supported this possibility. Purification of the sheath-degrading enzyme was carried out from the culture broth of strain TB. The molecular weight of the enzyme was calculated to be 78,000 and 50, 000 by sodium dodecyl sulfate-polyacrylamide electrophoresis and gel filtration chromatography, respectively. Enzyme activity was optimized at pH 6.5 to 7.0 and 30 to 40 degrees C. The reaction was accelerated by the addition of Mg(2+), Ca(2+), Fe(3+), and iodoacetamide, whereas it was inhibited by the addition of Cu(2+), Mn(2+), and dithiothreitol. The enzyme acted on the polysaccharide moiety of the sheath, producing an oligosaccharide the size of which was between the sizes of maltopentaose and maltohexaose. As the reaction proceeded, the absorbance at 235 nm of the reaction mixture increased, suggesting the generation of unsaturated sugars. Incorporation of unsaturated sugars was also suggested by the thiobarbituric acid reaction. It is possible that the enzyme is not a hydrolytic enzyme but a kind of polysaccharide eliminase which acts on the basic polysaccharide. PMID- 11055956 TI - Transition from anaerobic to aerobic growth conditions for the sulfate-reducing bacterium Desulfovibrio oxyclinae results in flocculation. AB - A chemostat culture of the sulfate-reducing bacterium Desulfovibrio oxyclinae isolated from the oxic layer of a hypersaline cyanobacterial mat was grown anaerobically and then subjected to gassing with 1% oxygen, both at a dilution rate of 0.05 h(-1). The sulfate reduction rate under anaerobic conditions was 370 nmol of SO(4)(2-) mg of protein(-1) min(-1). At the onset of aerobic gassing, sulfate reduction decreased by 40%, although viable cell numbers did not decrease. After 42 h, the sulfate reduction rate returned to the level observed in the anaerobic culture. At this stage the growth yield increased by 180% compared to the anaerobic culture to 4.4 g of protein per mol of sulfate reduced. Protein content per cell increased at the same time by 40%. The oxygen consumption rate per milligram of protein measured in washed cell suspensions increased by 80%, and the thiosulfate reduction rate of the same samples increased by 29% with lactate as the electron donor. These findings indicated possible oxygen-dependent enhancement of growth. After 140 h of growth under oxygen flux, formation of cell aggregates 0.1 to 3 mm in diameter was observed. Micrometer-sized aggregates were found to form earlier, during the first hours of exposure to oxygen. The respiration rate of D. oxyclinae was sufficient to create anoxia inside clumps larger than 3 microm, while the levels of dissolved oxygen in the growth vessel were 0.7 +/- 0.5 microM. Aggregation of sulfate-reducing bacteria was observed within a Microcoleus chthonoplastes-dominated layer of a cyanobacterial mat under daily exposure to oxygen concentrations of up to 900 microM. Desulfonema-like sulfate-reducing bacteria were also common in this environment along with other nonaggregated sulfate-reducing bacteria. Two dimensional mapping of sulfate reduction showed heterogeneity of sulfate reduction activity in this oxic zone. PMID- 11055957 TI - Sulfate reduction and possible aerobic metabolism of the sulfate-reducing bacterium Desulfovibrio oxyclinae in a chemostat coculture with Marinobacter sp. Strain MB under exposure to increasing oxygen concentrations. AB - A chemostat coculture of the sulfate-reducing bacterium Desulfovibrio oxyclinae together with a facultative aerobe heterotroph tentatively identified as Marinobacter sp. strain MB was grown under anaerobic conditions and then exposed to a stepwise-increasing oxygen influx (0 to 20% O(2) in the incoming gas phase). The coculture consumed oxygen efficiently, and no residual oxygen was detected with an oxygen supply of up to 5%. Sulfate reduction persisted at all levels of oxygen input, even at the maximal level, when residual oxygen in the growth vessel was 87 microM. The portion of D. oxyclinae cells in the coculture decreased gradually from 92% under anaerobic conditions to 27% under aeration. Both absolute cell numbers and viable cell counts of the organism were the same as or even higher than those observed in the absence of oxygen input. The patterns of consumption of electron donors and acceptors suggest that aerobic incomplete oxidation of lactate to acetate is performed by D. oxyclinae under high oxygen input. Both organisms were isolated from the same oxic zone of a cyanobacterial mat where they have to adapt to daily shifts from oxic to anoxic conditions. This type of syntrophic association may occur in natural habitats, enabling sulfate-reducing bacteria to cope with periodic exposure to oxygen. PMID- 11055958 TI - Oxygen-dependent growth of the sulfate-reducing bacterium Desulfovibrio oxyclinae in coculture with Marinobacter sp. Strain MB in an aerated sulfate-depleted chemostat. AB - A chemostat coculture of the sulfate-reducing bacterium Desulfovibrio oxyclinae and the facultatively aerobic heterotroph Marinobacter sp. strain MB was grown for 1 week under anaerobic conditions at a dilution rate of 0.05 h(-1). It was then exposed to an oxygen flux of 223 micromol min(-1) by gassing the growth vessel with 5% O(2). Sulfate reduction persisted under these conditions, though the amount of sulfate reduced decreased by 45% compared to the amount reduced during the initial anaerobic mode. After 1 week of growth under these conditions, sulfate was excluded from the incoming medium. The sulfate concentration in the growth vessel decreased exponentially from 4.1 mM to 2.5 microM. The coculture consumed oxygen effectively, and no residual oxygen was detected during either growth mode in which oxygen was supplied. The proportion of D. oxyclinae cells in the coculture as determined by in situ hybridization decreased from 86% under anaerobic conditions to 70% in the microaerobic sulfate-reducing mode and 34% in the microaerobic sulfate-depleted mode. As determined by the most-probable-number (MPN) method, the numbers of viable D. oxyclinae cells during the two microaerobic growth modes decreased compared to the numbers during the anaerobic growth mode. However, there was no significant difference between the MPN values for the two modes when oxygen was supplied. The patterns of consumption of electron donors and acceptors suggested that when oxygen was supplied in the absence of sulfate and thiosulfate, D. oxyclinae performed incomplete aerobic oxidation of lactate to acetate. This is the first observation of oxygen dependent growth of a sulfate-reducing bacterium in the absence of either sulfate or thiosulfate. Cells harvested during the microaerobic sulfate-depleted stage and exposed to sulfate and thiosulfate in a respiration chamber were capable of anaerobic sulfate and thiosulfate reduction. PMID- 11055959 TI - Specific secretion of active single-chain Fv antibodies into the supernatants of Escherichia coli cultures by use of the hemolysin system. AB - A simple method for the nontoxic, specific, and efficient secretion of active single-chain Fv antibodies (scFvs) into the supernatants of Escherichia coli cultures is reported. The method is based on the well-characterized hemolysin transport system (Hly) of E. coli that specifically secretes the target protein from the bacterial cytoplasm into the extracellular medium without a periplasmic intermediate. The culture media that accumulate these Hly-secreted scFv's can be used in a variety of immunoassays without purification. In addition, these culture supernatants are stable over long periods of time and can be handled basically as immune sera. PMID- 11055960 TI - Colonization of the stratified squamous epithelium of the nonsecreting area of horse stomach by lactobacilli. AB - Selective adhesion to only certain epithelia is particularly common among the bacterial members of the indigenous microflora of mammals. We have found that the stratified squamous epithelium of the nonsecreting area of horse stomach is colonized by gram-positive rods. The microscopic features of a dense layer of these bacteria on the epithelium were found to be similar to those reported in mice, rats, and swine. Adhering microorganisms were isolated and identified as Lactobacillus salivarius, L. crispatus, L. reuteri, and L. agilis by DNA-DNA hybridization and 16S rRNA gene sequencing techniques. These lactobacilli associated with the horse, except for L. reuteri, were found to adhere to horse epithelial cells in vitro but not to those of rats. A symbiotic relationship of these lactobacilli with the horse is suggested. PMID- 11055961 TI - Phylogenetic characterization and in situ detection of a Cytophaga-Flexibacter Bacteroides phylogroup bacterium in Tuber borchii vittad. Ectomycorrhizal mycelium. AB - Mycorrhizal ascomycetous fungi are obligate ectosymbionts that colonize the roots of gymnosperms and angiosperms. In this paper we describe a straightforward approach in which a combination of morphological and molecular methods was used to survey the presence of potentially endo- and epiphytic bacteria associated with the ascomycetous ectomycorrhizal fungus Tuber borchii Vittad. Universal eubacterial primers specific for the 5' and 3' ends of the 16S rRNA gene (16S rDNA) were used for PCR amplification, direct sequencing, and phylogenetic analyses. The 16S rDNA was amplified directly from four pure cultures of T. borchii Vittad. mycelium. A nearly full-length sequence of the gene coding for the prokaryotic small-subunit rRNA was obtained from each T. borchii mycelium studied. The 16S rDNA sequences were almost identical (98 to 99% similarity), and phylogenetic analysis placed them in a single unique rRNA branch belonging to the Cytophaga-Flexibacter-Bacteroides (CFB) phylogroup which had not been described previously. In situ detection of the CFB bacterium in the hyphal tissue of the fungus T. borchii was carried out by using 16S rRNA-targeted oligonucleotide probes for the eubacterial domain and the Cytophaga-Flexibacter phylum, as well as a probe specifically designed for the detection of this mycelium-associated bacterium. Fluorescent in situ hybridization showed that all three of the probes used bound to the mycelium tissue. This study provides the first direct visual evidence of a not-yet-cultured CFB bacterium associated with a mycorrhizal fungus of the genus Tuber. PMID- 11055962 TI - Phylogenetic analysis of and oligonucleotide probe development for eikelboom type 021N filamentous bacteria isolated from bulking activated sludge. AB - Fifteen filamentous strains, morphologically classified as Eikelboom type 021N bacteria, were isolated from bulking activated sludges. Based on comparative 16S ribosomal DNA (rDNA) sequence analysis, all strains form a monophyletic cluster together with all recognized Thiothrix species (88.3 to 98.7% 16S rDNA sequence similarity) within the gamma-subclass of Proteobacteria. The investigated Eikelboom type 021N isolates were subdivided into three distinct groups (I to III) demonstrating a previously unrecognized genetic diversity hidden behind the uniform morphology of the filaments. For in situ detection of these bacteria, 16S rRNA-targeted oligonucleotide probes specific for the entire Eikelboom type 021N Thiothrix cluster and the Eikelboom type 021N groups I, II, and III, respectively, were designed, evaluated, and successfully applied in activated sludge. PMID- 11055963 TI - Comparative 16S rRNA analysis of lake bacterioplankton reveals globally distributed phylogenetic clusters including an abundant group of actinobacteria. AB - In a search for cosmopolitan phylogenetic clusters of freshwater bacteria, we recovered a total of 190 full and partial 16S ribosomal DNA (rDNA) sequences from three different lakes (Lake Gossenkollesee, Austria; Lake Fuchskuhle, Germany; and Lake Baikal, Russia). The phylogenetic comparison with the currently available rDNA data set showed that our sequences fall into 16 clusters, which otherwise include bacterial rDNA sequences of primarily freshwater and soil, but not marine, origin. Six of the clusters were affiliated with the alpha, four were affiliated with the beta, and one was affiliated with the gamma subclass of the Proteobacteria; four were affiliated with the Cytophaga-Flavobacterium Bacteroides group; and one was affiliated with the class Actinobacteria (formerly known as the high-G+C gram-positive bacteria). The latter cluster (hgcI) is monophyletic and so far includes only sequences directly retrieved from aquatic environments. Fluorescence in situ hybridization (FISH) with probes specific for the hgcI cluster showed abundances of up to 1.7 x 10(5) cells ml(-1) in Lake Gossenkollesee, with strong seasonal fluctuations, and high abundances in the two other lakes investigated. Cell size measurements revealed that Actinobacteria in Lake Gossenkollesee can account for up to 63% of the bacterioplankton biomass. A combination of phylogenetic analysis and FISH was used to reveal 16 globally distributed sequence clusters and to confirm the broad distribution, abundance, and high biomass of members of the class Actinobacteria in freshwater ecosystems. PMID- 11055964 TI - Rapid detection and quantification of members of the archaeal community by quantitative PCR using fluorogenic probes. AB - We describe a rapid, reproducible, and sensitive method for detection and quantification of archaea in naturally occurring microbial communities. A domain specific PCR primer set and a domain-specific fluorogenic probe having strong and weak selectivity, respectively, for archaeal rRNA genes (rDNAs) were designed. A universal PCR primer set and a universal fluorogenic probe for both bacterial and archaeal rDNAs were also designed. Using these primers and probes, we demonstrated that detection and quantification of archaeal rDNAs in controlled microbial rDNA assemblages can be successfully achieved. The system which we designed was also able to detect and quantify archaeal rDNAs in DNA samples obtained not only from environments in which thermophilic archaea are abundant but also from environments in which methanogenic archaea are abundant. Our findings indicate that this method is applicable to culture-independent molecular analysis of microbial communities in various environments. PMID- 11055965 TI - Relationships between colony morphotypes and oil tolerance in Rhodococcus rhodochrous. AB - A mucoidal strain of Rhodococcus rhodochrous was resistant to 10% (vol/vol) n hexadecane, while its rough derivatives were sensitive. When the extracellular polysaccharide (EPS) produced by the mucoidal strain was added to cultures of the rough strains, the rough strains gained resistance to n-hexadecane. Thus, EPS confer tolerance to n-hexadecane in members of the genus Rhodococcus. PMID- 11055966 TI - Bradyrhizobium sp. Strains that nodulate the leguminous tree Acacia albida produce fucosylated and partially sulfated nod factors. AB - We determined the structures of Nod factors produced by six different Bradyrhizobium sp. strains nodulating the legume tree Acacia albida (syn. Faidherbia albida). Compounds from all strains were found to be similar, i.e., O carbamoylated and substituted by an often sulfated methyl fucose and different from compounds produced by Rhizobium-Mesorhizobium-Sinorhizobium strains nodulating other species of the Acaciae tribe. PMID- 11055967 TI - Low sensitivity of Listeria monocytogenes to quaternary ammonium compounds. AB - Ninety-seven epidemiologically unrelated strains of Listeria monocytogenes were investigated for their sensitivities to quaternary ammonium compounds (benzalkonium chloride and cetrimide). The MICs for seven serogroup 1/2 strains were high. Three came from the environment and four came from food; none were isolated from human or animal samples. All 97 strains carried the mdrL gene, which encodes a multidrug efflux pump, and the orfA gene, a putative transcriptional repressor of mdrL. The absence of plasmids in four of the seven resistant strains and the conservation of resistance after plasmid curing suggested that the resistance genes are not plasmid borne. Moreover, PCR amplification and Southern blot hybridization experiments failed to find genes phylogenetically related to the qacA and smr genes, encoding multidrug efflux systems previously described for the genus Staphylococcus. The high association between nontypeability by phages and the loss of sensitivity to quaternary ammonium compounds are suggestive of an intrinsic resistance due to modifications in the cell wall. PMID- 11055968 TI - Assessment of poliovirus eradication in Japan: genomic analysis of polioviruses isolated from river water and sewage in toyama prefecture. AB - Seventy-eight poliovirus strains isolated from river water and sewage in Toyama Prefecture, Japan, during 1993 to 1995 were characterized by the PCR-restriction fragment length polymorphism (RFLP) method and by partially sequencing the VP3 and VP1 regions of the viral genome. Of these isolates, 36 were identified as Sabin vaccine strains, and 42 were identified as vaccine variant strains that had less than 1.4% nucleotide divergence from the Sabin strains, including 7 isolates with patterns different from those of Sabin strains as determined by PCR-RFLP analysis. These findings suggest that wild-type poliovirus was not circulating in Toyama Prefecture. PMID- 11055969 TI - Heavy metal coprecipitation with hydrozincite [Zn(5)(CO(3))(2)(OH)(6)] from mine waters caused by photosynthetic microorganisms. AB - An iron-poor stream of nearly neutral pH polluted by mine tailings has been investigated for a natural phenomenon responsible for the polishing of heavy metals in mine wastewaters. A white mineralized mat, which was determined to be hydrozincite [Zn(5)(CO(3))(2)(OH)(6)] by X-ray diffraction analysis, was observed in the stream sediments mainly in spring. The precipitate shows a total organic matter residue of 10% dry weight and contains high concentrations of Pb, Cd, Ni, Cu, and other metals. Scanning electron microscopy analysis suggests that hydrozincite is mainly of biological origin. Dormant photosynthetic microorganisms have been retrieved from 1-year-old dry hydrozincite. The autofluorescent microorganisms were imaged by a scanning confocal laser microscope. A photosynthetic filamentous bacterium, classified as Scytonema sp. strain ING-1, was found associated with microalga Chlorella sp. strain SA1. This microbial community is responsible for the natural polishing of heavy metals in the water stream by coprecipitation with hydrozincite. PMID- 11055970 TI - Genetic characterization of soybean rhizobia in Paraguay. AB - The soybean is an exotic plant introduced in Paraguay in this century; commercial cropping expanded after the 1970s. Inoculation is practiced in just 15 to 20% of the cropping areas, but root nodulation occurs in most sites where soybeans grow. Little is known about rhizobial diversity in South America, and no study has been performed in Paraguay until this time. Therefore, in this study, the molecular characterization of 78 rhizobial isolates from soybean root nodules, collected under field conditions in 16 sites located in the two main producing states, Alto Parana and Itapua, was undertaken. A high level of genetic diversity was detected by an ERIC-REP-PCR analysis, with the majority of the isolates representing unique strains. Most of the 58 isolates characterized by slow growth and alkaline reactions in a medium containing mannitol as a carbon source were clustered with strains representative of the Bradyrhizobium japonicum and Bradyrhizobium elkanii species, and the 16S ribosomal DNA (rDNA) sequences of 5 of those isolates confirmed the species identities. However, slow growers were highly polymorphic in relation to the reference strains, including five carried in commercial inoculants in neighboring countries, thus indicating that the Paraguayan isolates might represent native bradyrhizobia. Twenty isolates highly polymorphic in the ERIC-REP-PCR profiles were characterized by fast growth and acid reactions in vitro, and two of them showed high 16S rDNA identities with Rhizobium genomic species Q. However, two other fast growers showed high 16S rDNA identity with Agrobacterium spp., and both of these strains established efficient symbioses with soybean plants. PMID- 11055971 TI - Adventure of gene therapy into the brain: A new era for cardiovascular gene therapy. PMID- 11055972 TI - Hyperinsulinemia and vascular dysfunction: the role of nuclear factor-kappaB, yet again. PMID- 11055973 TI - When does spontaneous sarcoplasmic reticulum CA(2+) release cause a triggered arrythmia? Cellular versus tissue requirements. PMID- 11055974 TI - Endothelial cells of hematopoietic origin make a significant contribution to adult blood vessel formation. AB - Granulation tissue formation is an example of new tissue development in an adult. Its rich vascular network has been thought to derive via angiogenic sprouting and extension of preexisting vessels from the surrounding tissue. The possibility that circulating cells of hematopoietic origin can differentiate into vascular endothelial cells (ECs) in areas of vascular remodeling has recently gained credibility. However, no quantitative data have placed the magnitude of this contribution into a physiological perspective. We have used hematopoietic chimeras to determine that 0.2% to 1.4% of ECs in vessels in control tissues derived from hematopoietic progenitors during the 4 months after irradiation and hematopoietic recovery. By contrast, 8.3% to 11.2% of ECs in vessels that developed in sponge-induced granulation tissue during 1 month derived from circulating hematopoietic progenitors. This recruitment of circulating progenitors to newly forming vessels would be difficult to observe in standard histological studies, but it is large enough to be encouraging for attempts to manipulate this contribution for therapeutic gain. PMID- 11055975 TI - Calcineurin and beyond: cardiac hypertrophic signaling. AB - In response to increased ventricular wall tension or neurohumoral stimuli, the myocardium undergoes an adaptive hypertrophy response that temporarily augments pump function. Although initially beneficial, sustained cardiac hypertrophy can lead to decompensation and cardiomyopathy. Recent studies have focused on characterizing the molecular mechanisms that underlie cardiac hypertrophy. An increasing number of signal transduction pathways have been identified as important regulators of the hypertrophic response, including the low-molecular weight GTPases (Ras, RhoA, and Rac), mitogen-activated protein kinases, protein kinase C, and calcineurin. This review will discuss an emerging body of evidence that implicates the calcium-calmodulin-activated protein phosphatase calcineurin as a physiological regulator of the cardiac hypertrophic response. Although the sufficiency of calcineurin to promote cardiomyocyte hypertrophy in vivo and in vitro is established, its overall necessity as a hypertrophic mediator is currently an area of ongoing debate. The use of the calcineurin-inhibitory agents cyclosporine A and FK506 have suggested a necessary role for calcineurin in many, but not all, animal models of hypertrophy or cardiomyopathy. The evidence implicating a role for calcineurin signaling in the heart will be weighed against a growing body of literature suggesting necessary roles for a diverse array of intracellular signaling pathways, highlighting the multifactorial nature of the hypertrophic program. PMID- 11055976 TI - Reversal of angiogenesis in vitro, induction of apoptosis, and inhibition of AKT phosphorylation in endothelial cells by thromboxane A(2). AB - Thromboxane A(2) (TxA(2)) causes platelet aggregation, vasoconstriction, and inhibition of endothelial cell (EC) migration and prevents vascular tube formation via its specific receptors (TP), of which there are two isoforms (TPalpha and TPbeta), both expressed in human ECs. In this study, we demonstrate that the TxA(2) mimetic IBOP increases apoptosis of human ECs and inhibits the phosphorylation of Akt kinase, an intracellular mediator required for cell survival. Treatment with IBOP destroyed EC networks formed on a basement membrane matrix in vitro. To distinguish the role of the TP isoforms, each isoform was expressed in TP-null ECs to create TPalpha and TPbeta ECs. IBOP induced apoptosis and inhibited phosphorylation of Akt kinase in both TPalpha and TPbeta. IBOP increased cAMP levels in TPalpha but not in TPbeta. Apoptosis induced by IBOP in TPalpha was not affected by either the adenylyl cyclase activator forskolin or the protein kinase A inhibitor 14-22 amide or H-89, whereas that in TPbeta was suppressed by forskolin and enhanced by the protein kinase A inhibitor 14-22 amide or H-89, suggesting that the TP isoforms differ in their signal pathways in mediating apoptosis. In conclusion, apoptosis may be the mechanism by which TxA(2)-mediated destruction of vascular structures in ECs occurs; although both TP isoforms induce apoptosis, possibly via inhibiting Akt phosphorylation, the signaling differs in each isoform, in that activation of the adenylyl cyclase pathway prevents apoptosis caused by TPbeta, but not by TPalpha, stimulation. PMID- 11055977 TI - Hyperinsulinemia enhances transcriptional activity of nuclear factor-kappaB induced by angiotensin II, hyperglycemia, and advanced glycosylation end products in vascular smooth muscle cells. AB - Pathogenesis of macrovascular complications of diabetes may involve an activation of the transcription factor nuclear factor-kappaB (NF-kappaB) by hyperglycemia and advanced glycosylation end products (AGEs). Activation of NF-kappaB is believed to be dependent on activation of the Rho family of GTPases. Although the precise mechanism of the Rho-mediated action is not completely understood, posttranslational modification of the Rho proteins by geranylgeranylation is required for their subsequent activation. We observed that in cultured vascular smooth muscle cells (VSMCs), insulin stimulated the activity of geranylgeranyltransferase (GGTase) I and increased the amounts of geranylgeranylated Rho-A from 47% to 60% (P:<0.05). GGTI-286, an inhibitor of GGTase I, blocked both effects of insulin. Increased availability of prenylated Rho-A significantly augmented the abilities of angiotensin II (Ang II), hyperglycemia, and AGEs to activate NF-kappaB, as measured by NF-kappaB response element luciferase reporter activity. Preincubations of VSMCs with insulin for 24 hours doubled NF-kappaB transactivation by Ang II, hyperglycemia, and AGEs. This priming effect of insulin was completely inhibited by GGTI-286. We demonstrate for the first time, to our knowledge, that insulin potentiates NF-kappaB dependent transcriptional activity induced by hyperglycemia, AGEs, and Ang II in VSMCs by increasing the activity of GGTase I and the availability of geranylgeranylated Rho-A. PMID- 11055978 TI - Functional reconstitution of the angiotensin II type 2 receptor and G(i) activation. AB - On the basis of the patterns of conserved amino acid sequence, the angiotensin II type 2 (AT(2)) receptor belongs to the family of serpentine receptors, which relay signals from extracellular stimuli to heterotrimeric G proteins. However, the AT(2) receptor signal transduction mechanisms are poorly understood. We have measured AT(2)-triggered activation of purified heterotrimeric proteins in urea extracted membranes from cultured COS-7 cells expressing the recombinant receptor. This procedure removes contaminating GTP-binding proteins without inactivating the serpentine receptor. Binding studies using [(125)I] angiotensin (Ang) II revealed a single binding site with a K(d)=0.45 and a capacity of 627 fmol/mg protein in the extracted membranes. The AT(2) receptor caused a rapid activation of alpha(i) and alpha(o) but not of alpha(q) and alpha(s), as measured by radioactive guanosine 5'-3-O-(thio)triphosphate (GTPgammaS) binding. Activation required the presence of activated receptors, betagamma, and alpha subunits. As a first step aimed at developing an in vitro assay to examine AT(2) receptor pharmacology, we tested a battery of Ang II-related ligands for their ability to promote AT(1) or AT(2) receptor-catalyzed G(i) activation. Two proteolytic fragments of Ang II, Ang III and Ang1-7, also promoted activation of alpha(i) through the AT(2) receptor. Furthermore, we found that [Sar(1),Ala(8)]Ang II is an antagonist for both AT(1) and AT(2) receptors and that CPG42112 behaves as a partial agonist for the AT(2) receptor. In combination with previous observations, these results show that the AT(2) receptor is fully capable of activating G(i) and provides a new tool for exploring AT(2) receptor pharmacology and interactions with G-protein trimers. PMID- 11055979 TI - Sinoatrial node pacemaker activity requires Ca(2+)/calmodulin-dependent protein kinase II activation. AB - Cardiac beating arises from the spontaneous rhythmic excitation of sinoatrial (SA) node cells. Here we report that SA node pacemaker activity is critically dependent on Ca(2+)/calmodulin-dependent protein kinase II (CaMKII). In freshly dissociated rabbit single SA node cells, inhibition of CaMKII by a specific peptide inhibitor, autocamtide-2 inhibitory peptide (AIP, 10 micromol/L), or by KN-93 (0.1 to 3.0 micromol/L), but not its inactive analog, KN-92, depressed the rate and amplitude of spontaneous action potentials (APs) in a dose-dependent manner. Strikingly, 10 micromol/L AIP and 3 micromol/L KN-93 completely arrested SA node cells, which indicates that basal CaMKII activation is obligatory to the genesis of pacemaker AP. To understand the ionic mechanisms of the CaMKII effects, we measured L-type Ca(2+) current (I(Ca, L)), which contributes both to AP upstroke and to pacemaker depolarization. KN-93 (1 micromol/L), but not its inactive analog, KN-92, decreased I:(Ca, L) amplitude from 12+/-2 to 6+/-1 pA/pF without altering the shape of the current-voltage relationship. Both AIP and KN 93 shifted the midpoint of the steady-state inactivation curve leftward and markedly slowed the recovery of I(Ca, L) from inactivation. Similar results were observed using the fast Ca(2+) chelator BAPTA, whereas the slow Ca(2+) chelator EGTA had no significant effect, which suggests that CaMKII activity is preferentially regulated by local Ca(2+) transients. Indeed, confocal immunocytochemical imaging showed that active CaMKII is highly localized beneath the surface membrane in the vicinity of L-type channels and that AIP and KN-93 significantly reduced CaMKII activity. Thus, we conclude that CaMKII plays a vital role in regulating cardiac pacemaker activity mainly via modulating I(Ca, L) inactivation and reactivation, and local Ca(2+) is critically involved in these processes. PMID- 11055980 TI - Strongly binding myosin crossbridges regulate loaded shortening and power output in cardiac myocytes. AB - This study investigated the possible roles of strongly binding myosin crossbridges in determining loaded shortening and power output in cardiac myocytes. Single skinned cardiac myocytes were attached between a force transducer and position motor, and shortening velocities were measured over a range of loads during varying levels of Ca(2+) activation. Lowering the [Ca(2+)] slowed shortening velocities, decreased relative power output, and increased the curvature of length traces. We tested the hypothesis that Ca(2+) activation dependence of loaded shortening is determined primarily by strongly binding crossbridges or by [Ca(2+)] per se, which was done by measuring loaded shortening before and after addition of N-ethylmaleimide-conjugated myosin subfragment-1 (NEM-S1), a strongly binding myosin analogue that cooperatively enhances thin filament activation. At fixed [Ca(2+)], NEM-S1 reduced the curvature of length traces and sped loaded shortening velocities. Even when [Ca(2+)] was adjusted so that force was equal with and without NEM-S1, myocyte shortening was faster and exhibited less curvature with NEM-S1. In the presence of NEM-S1, peak relative power output was also significantly greater during activations either at the same [Ca(2+)] or when [Ca(2+)] was adjusted to achieve the same force. Consequently, NEM-S1 eliminated any Ca(2+) dependence of relative power output that is normally observed in cardiac myocytes. These results indicate that strongly binding crossbridges play a significant role in determining loaded shortening and power output and suggest that previously observed Ca(2+) dependence of power output is mediated by alterations in numbers of crossbridges bound to the thin filament. PMID- 11055981 TI - Sarcoplasmic reticulum Ca(2+) release causes myocyte depolarization. Underlying mechanism and threshold for triggered action potentials. AB - Spontaneous sarcoplasmic reticulum (SR) Ca(2+) release causes delayed afterdepolarizations (DADs) via Ca(2+)-induced transient inward currents (I:(ti)). However, no quantitative data exists regarding (1) Ca(2+) dependence of DADs, (2) Ca(2+) required to depolarize the cell to threshold and trigger an action potential (AP), or (3) relative contributions of Ca(2+)-activated currents to DADs. To address these points, we evoked SR Ca(2+) release by rapid application of caffeine in indo 1-AM-loaded rabbit ventricular myocytes and measured caffeine-induced DADs (cDADs) with whole-cell current clamp. The SR Ca(2+) load of the myocyte was varied by different AP frequencies. The cDAD amplitude doubled for every 88+/-8 nmol/L of Delta[Ca(2+)](i) (simple exponential), and the Delta[Ca(2+)](i) threshold of 424+/-58 nmol/L was sufficient to trigger an AP. Blocking Na(+)-Ca(2+) exchange current (I(Na/Ca)) by removal of [Na](o) and [Ca(2+)](o) (or with 5 mmol/L Ni(2+)) reduced cDADs by >90%, for the same Delta[Ca(2+)](i). In contrast, blockade of Ca(2+)-activated Cl(-) current (I(Cl(Ca))) with 50 micromol/L niflumate did not significantly alter cDADs. We conclude that DADs are almost entirely due to I(Na/Ca), not I(Cl(Ca)) or Ca(2+)-activated nonselective cation current. To trigger an AP requires 30 to 40 micromol/L cytosolic Ca(2+) or a [Ca(2+)](i) transient of 424 nmol/L. Current injection, simulating I(ti)s with different time courses, revealed that faster I:(ti)s require less charge for AP triggering. Given that spontaneous SR Ca(2+) release occurs in waves, which are slower than cDADs or fast I(ti)s, the true Delta[Ca(2+)](i) threshold for AP activation may be approximately 3-fold higher in normal myocytes. This provides a safety margin against arrhythmia in normal ventricular myocytes. PMID- 11055982 TI - Autonomous and growth factor-induced hypertrophy in cultured neonatal mouse cardiac myocytes. Comparison with rat. AB - Cultured neonatal rat cardiac myocytes have been used extensively to study cellular and molecular mechanisms of cardiac hypertrophy. However, there are only a few studies in cultured mouse myocytes despite the increasing use of genetically engineered mouse models of cardiac hypertrophy. Therefore, we characterized hypertrophic responses in low-density, serum-free cultures of neonatal mouse cardiac myocytes and compared them with rat myocytes. In mouse myocyte cultures, triiodothyronine (T3), norepinephrine (NE) through a beta adrenergic receptor, and leukemia inhibitory factor induced hypertrophy by a 20% to 30% increase in [(3)H]phenylalanine-labeled protein content. T3 and NE also increased alpha-myosin heavy chain (MyHC) mRNA and reduced beta-MyHC. In contrast, hypertrophic stimuli in rat myocytes, including alpha(1)-adrenergic agonists, endothelin-1, prostaglandin F(2alpha), interleukin 1beta, and phorbol 12-myristate 13-acetate (PMA), had no effect on mouse myocyte protein content. In further contrast with the rat, none of these agents increased atrial natriuretic factor or beta-MyHC mRNAs. Acute PMA signaling was intact by extracellular signal regulated kinase (ERK1/2) and immediate-early gene (fos/jun) activation. Remarkably, mouse but not rat myocytes had hypertrophy in the absence of added growth factors, with increases in cell area, protein content, and the mRNAs for atrial natriuretic factor and beta-MyHC. We conclude that mouse myocytes have a unique autonomous hypertrophy. On this background, T3, NE, and leukemia inhibitory factor activate hypertrophy with different mRNA phenotypes, but certain Gq- and protein kinase C-coupled agonists do not. PMID- 11055983 TI - Cyclophilin A is a secreted growth factor induced by oxidative stress. AB - Reactive oxygen species have been implicated in the pathogenesis of atherosclerosis, hypertension, and restenosis, in part by promoting vascular smooth muscle cell (VSMC) growth. Many VSMC growth factors are secreted by VSMC and act in an autocrine manner. Here we demonstrate that cyclophilin A (CyPA), a member of the immunophilin family, is secreted by VSMCs in response to oxidative stress and mediates extracellular signal-regulated kinase (ERK1/2) activation and VSMC growth by reactive oxygen species. Human recombinant CyPA can mimic the effects of secreted CyPA to stimulate ERK1/2 and cell growth. The peptidyl-prolyl isomerase activity is required for ERK1/2 activation by CyPA. In vivo, CyPA expression and secretion are increased by oxidative stress and vascular injury. These findings are the first to identify CyPA as a secreted redox-sensitive mediator, establish CyPA as a VSMC growth factor, and suggest an important role for CyPA and enzymes with peptidyl-prolyl isomerase activity in the pathogenesis of vascular diseases. PMID- 11055984 TI - The role of electroporation in defibrillation. AB - Electric shock is the only effective therapy against ventricular fibrillation. However, shocks are also known to cause electroporation of cell membranes. We sought to determine the impact of electroporation on ventricular conduction and defibrillation. We optically mapped electrical activity in coronary-perfused rabbit hearts during electric shocks (50 to 500 V). Electroporation was evident from transient depolarization, reduction of action potential amplitude, and upstroke dV/dt. Electroporation was voltage dependent and significantly more pronounced at the endocardium versus the epicardium, with thresholds of 229+/-81 versus 318+/-84 V, respectively (P=0.01, n=10), both being above the defibrillation threshold of 181.3+/-45.8 V. Epicardial electroporation was localized to a small area near the electrode, whereas endocardial electroporation was observed at the bundles and trabeculas throughout the entire endocardium. Higher-resolution imaging revealed that papillary muscles (n=10) were most affected. Electroporation and conduction block thresholds in papillary muscles were 281+/-64 V and 380+/-79 V, respectively. We observed no arrhythmia in association with electroporation. Further, preconditioning with high-energy shocks prevented reinduction of fibrillation by 50-V shocks, which were otherwise proarrhythmic. Endocardial bundles are the most susceptible to electroporation and the resulting conduction impairment. Electroporation is not associated with proarrhythmic effects and is associated with a reduction of vulnerability. PMID- 11055985 TI - Transgenic modeling of a cardiac troponin I mutation linked to familial hypertrophic cardiomyopathy. AB - Multiple mutations in cardiac troponin I (cTnI) have been associated with familial hypertrophic cardiomyopathy. Two mutations are located in the cTnI inhibitory domain, a highly negatively charged region that alternately binds to either actin or troponin C, depending on the intracellular concentration of calcium. This region is critical to the inhibition of actin-myosin crossbridge formation when intracellular calcium is low. We modeled one of the inhibitory domain mutations, arginine145-->glycine (TnI(146Gly) in the mouse sequence), by cardiac-specific expression of the mutated protein in transgenic mice. Multiple lines were generated with varying degrees of expression to establish a dose relationship; the severity of phenotype could be correlated directly with transgene expression levels. Transgenic mice overexpressing wild-type cTnI were generated as controls and analyzed in parallel with the TnI(146Gly) animals. The control mice showed no abnormalities, indicating that the phenotype of TnI(146Gly) was not simply an artifact of transgenesis. In contrast, TnI(146Gly) mice showed cardiomyocyte disarray and interstitial fibrosis and suffered premature death. The functional alterations that seem to be responsible for the development of cardiac disease include increased skinned fiber sensitivity to calcium and, at the whole organ level, hypercontractility with diastolic dysfunction. Severely affected lines develop a pathology similar to human familial hypertrophic cardiomyopathy but within a dramatically shortened time frame. These data establish the causality of this mutation for cardiac disease, provide an animal model for understanding the resultant pathogenic structure function relationships, and highlight the differences in phenotype severity of the troponin mutations between human and mouse hearts. PMID- 11055986 TI - Myocardial ischemia/reperfusion injury in NADPH oxidase-deficient mice. AB - Previous studies have suggested that oxygen-derived free radicals are involved in the pathophysiology of myocardial ischemia/reperfusion (MI/R) injury. Specifically, neutrophils have been shown to mediate postischemic ventricular arrhythmias and myocardial necrosis. We hypothesized that MI/R injury would be reduced in the absence (-/-) of NADPH oxidase. Heterozygous control mice (n=23) and NADPH oxidase(-/-) mice (n=24) were subjected to 30 minutes of coronary artery occlusion and 24 hours of reperfusion. Myocardial area at risk per left ventricle was similar in heterozygous control hearts (55+/-3%) and NADPH oxidase( /-) hearts (61+/-4%). Contrary to our hypothesis, the size of infarct area at risk was similar in the heterozygous control mice (42+/-4%) and NADPH oxidase(-/ ) mice (34+/-5%) (P=not significant). In addition, echocardiographic examination of both groups revealed that left ventricle fractional shortening was similar in NADPH oxidase(-/-) mice (n=8; 27+/-2.5%) and heterozygous control mice (n=10; 23.3+/-3. 3%) after MI/R. Superoxide production, as detected by cytochrome c reduction, was significantly impaired (P<0.01) in NADPH oxidase(-/-) mice (n=6) compared with heterozygous mice (n=7) (0.04+/-0.03 versus 2.2+/-0.08 nmol O(2).min(-1).10(6) cells(-1)). Intravital microscopy of the inflamed mesenteric microcirculation demonstrated that leukocyte rolling and adhesion were unaffected by the absence of NADPH oxidase. Oyster glycogen-stimulated neutrophil transmigration into the peritoneum was also similar in both the heterozygous control mice and NADPH oxidase(-/-) mice (P:=not significant). These findings suggest that NADPH oxidase does not contribute to the development of myocardial injury and dysfunction after MI/R. PMID- 11055987 TI - Gene transfer of calcitonin gene-related peptide prevents vasoconstriction after subarachnoid hemorrhage. AB - We sought to determine whether adenovirus-mediated gene transfer in vivo of calcitonin gene-related peptide (CGRP), a potent vasodilator, ameliorates cerebral vasoconstriction after experimental subarachnoid hemorrhage (SAH). Arterial blood was injected into the cisterna magna of rabbits to mimic SAH 5 days after injection of AdRSVCGRP (8x10(8) pfu), AdRSVbetagal (control virus), or vehicle. After injection of AdRSVCGRP, there was a 400-fold increase in CGRP in cerebrospinal fluid. Contraction of the basilar artery to serotonin in vitro was greater in rabbits after SAH than after injection of artificial cerebrospinal fluid (P<0.001). Contraction to serotonin was less in rabbits with SAH after AdRSVCGRP than after AdRSVbetagal or vehicle (P:<0.02). Basal diameter of the basilar artery before SAH (measured with digital subtraction angiogram) was 13% greater in rabbits treated with AdRSVCGRP than in rabbits treated with vehicle or AdRSVbetagal (P:<0.005). In rabbits treated with vehicle or AdRSVbetagal, arterial diameter after SAH was 25+/-3% smaller than before SAH (P<0.0005). In rabbits treated with AdRSVCGRP, arterial diameter was similar before and after SAH and was reduced by 19+/-3% (P<0.01) after intracisternal injection of CGRP-(8 37) (0.5 nmol/kg), a CGRP(1) receptor antagonist. To determine whether gene transfer of CGRP after SAH may prevent cerebral vasoconstriction, we constructed a virus with a cytomegalovirus (CMV) promoter, which results in rapid expression of the transgene product. Treatment of rabbits with AdCMVCGRP after experimental SAH prevented constriction of the basilar artery 2 days after SAH. Thus, gene transfer of CGRP prevents cerebral vasoconstriction in vivo after experimental SAH. PMID- 11055988 TI - Mechanisms of NO/cGMP-dependent vasorelaxation. AB - Both cGMP-dependent and -independent mechanisms have been implicated in the regulation of vascular tone by NO. We analyzed acetylcholine (ACh)- and NO induced relaxation in pressurized small arteries and aortic rings from wild-type (wt) and cGMP kinase I-deficient (cGKI(-/-)) mice. Low concentrations of NO and ACh decreased the spontaneous myogenic tone in wt but not in cGKI(-/-) arteries. However, contractions of cGKI(-/-) arteries and aortic rings were reduced by high concentrations (10 micromol/L) of 2-(N:, N-diethylamino)-diazenolate-2-oxide (DEA NO). Iberiotoxin, a specific blocker of Ca(2+)-activated K(+) (BK(Ca)) channels, only partially prevented the relaxation induced by DEA-NO or ACh in pressurized vessels and aortic rings. DEA-NO increased the activity of BK(Ca) channels only in vascular smooth muscle cells isolated from wt cGKI(+/+) mice. These results suggest that low physiological concentrations of NO decrease vascular tone through activation of cGKI, whereas high concentrations of DEA-NO relax vascular smooth muscle independent of cGKI and BK(Ca). NO-stimulated, cGKI-independent relaxation was antagonized by the inhibition of soluble guanylyl cyclase or cAMP kinase (cAK). DEA-NO increased cGMP to levels that are sufficient to activate cAK. cAMP-dependent relaxation was unperturbed in cGKI(-/-) vessels. In conclusion, low concentrations of NO relax vessels by activation of cGKI, whereas in the absence of cGKI, NO can relax small and large vessels by cGMP-dependent activation of cAK. PMID- 11055989 TI - Structural determinants of PIP(2) regulation of inward rectifier K(ATP) channels. AB - Phosphatidylinositol 4,5-bisphosphate (PIP(2)) activates K(ATP) and other inward rectifier (Kir) channels. To determine residues important for PIP(2) regulation, we have systematically mutated each positive charge in the COOH terminus of Kir6.2 to alanine. The effects of these mutations on channel function were examined using (86)Rb efflux assays on intact cells and inside-out patch-clamp methods. Both methods identify essentially the same basic residues in two narrow regions (176-222 and 301-314) in the COOH terminus that are important for the maintenance of channel function and interaction with PIP(2). Only one residue (R201A) simultaneously affected ATP and PIP(2) sensitivity, which is consistent with the notion that these ligands, while functionally competitive, are unlikely to bind to identical sites. Strikingly, none of 13 basic residues in the terminal portion (residues 315-390) of the COOH terminus affected channel function when neutralized. The data help to define the structural requirements for PIP(2) sensitivity of K(ATP) channels. Moreover, the regions and residues defined in this study parallel those uncovered in recent studies of PIP(2) sensitivity in other inward rectifier channels, indicating a common structural basis for PIP(2) regulation. PMID- 11055990 TI - On the mechanism of proton transport by the neuronal excitatory amino acid carrier 1. AB - Uptake of glutamate from the synaptic cleft is mediated by high affinity transporters and is driven by Na(+), K(+), and H(+) concentration gradients across the membrane. Here, we characterize the molecular mechanism of the intracellular pH change associated with glutamate transport by combining current recordings from excitatory amino acid carrier 1 (EAAC1)-expressing HEK293 cells with a rapid kinetic technique with a 100-micros time resolution. Under conditions of steady state transport, the affinity of EAAC1 for glutamate in both the forward and reverse modes is strongly dependent on the pH on the cis-side of the membrane, whereas the currents at saturating glutamate concentrations are hardly affected by the pH. Consistent with this, the kinetics of the pre-steady state currents, measured after saturating glutamate concentration jumps, are not a function of the pH. In addition, we determined the deuterium isotope effect on EAAC1 kinetics, which is in agreement with proton cotransport but not OH(-) countertransport. The results can be quantitatively explained with an ordered binding model that includes a rapid proton binding step to the empty transporter followed by glutamate binding and translocation of the proton-glutamate transporter complex. The apparent pK of the extracellular proton binding site is approximately 8. This value is shifted to approximately 6.5 when the substrate binding site is exposed to the cytoplasm. PMID- 11055991 TI - Molecular coupling of S4 to a K(+) channel's slow inactivation gate. AB - The mechanism by which physiological signals regulate the conformation of molecular gates that open and close ion channels is poorly understood. Voltage clamp fluorometry was used to ask how the voltage-sensing S4 transmembrane domain is coupled to the slow inactivation gate in the pore domain of the Shaker K(+) channel. Fluorophores attached at several sites in S4 indicate that the voltage sensing rearrangements are followed by an additional inactivation motion. Fluorophores attached at the perimeter of the pore domain indicate that the inactivation rearrangement projects from the selectivity filter out to the interface with the voltage-sensing domain. Some of the pore domain sites also sense activation, and this appears to be due to a direct interaction with S4 based on the finding that S4 comes into close enough proximity to the pore domain for a pore mutation to alter the nanoenvironment of an S4-attached fluorophore. We propose that activation produces an S4-pore domain interaction that disrupts a bond between the S4 contact site on the pore domain and the outer end of S6. Our results indicate that this bond holds the slow inactivation gate open and, therefore, we propose that this S4-induced bond disruption triggers inactivation. PMID- 11055992 TI - A hot spot for the interaction of gating modifier toxins with voltage-dependent ion channels. AB - The gating modifier toxins are a large family of protein toxins that modify either activation or inactivation of voltage-gated ion channels. omega-Aga-IVA is a gating modifier toxin from spider venom that inhibits voltage-gated Ca(2+) channels by shifting activation to more depolarized voltages. We identified two Glu residues near the COOH-terminal edge of S3 in the alpha(1A) Ca(2+) channel (one in repeat I and the other in repeat IV) that align with Glu residues previously implicated in forming the binding sites for gating modifier toxins on K(+) and Na(+) channels. We found that mutation of the Glu residue in repeat I of the Ca(2+) channel had no significant effect on inhibition by omega-Aga-IVA, whereas the equivalent mutation of the Glu in repeat IV disrupted inhibition by the toxin. These results suggest that the COOH-terminal end of S3 within repeat IV contributes to forming a receptor for omega-Aga-IVA. The strong predictive value of previous mapping studies for K(+) and Na(+) channel toxins argues for a conserved binding motif for gating modifier toxins within the voltage-sensing domains of voltage-gated ion channels. PMID- 11055993 TI - Single channel studies of inward rectifier potassium channel regulation by muscarinic acetylcholine receptors. AB - Negative regulation of the heartbeat rate involves the activation of an inwardly rectifying potassium current (I(KACh)) by G protein-coupled receptors such as the m2 muscarinic acetylcholine receptor. Recent studies have shown that this process involves the direct binding of G(betagamma) subunits to the NH(2)- and COOH terminal cytoplasmic domains of the proteins termed GIRK1 and GIRK4 (Kir3.1 and Kir3.4/CIR), which mediate I(KACh). Because of the very low basal activity of native I(KACh), it has been difficult to determine the single channel effect of G(betagamma) subunit binding on I(KACh) activity. Through analysis of a novel G protein-activated chimeric inward rectifier channel that displays increased basal activity relative to I(KACh), we find that single channel activation can be explained by a G protein-dependent shift in the equilibrium of open channel transitions in favor of a bursting state of channel activity over a long-lived closed state. PMID- 11055994 TI - A structural rearrangement in the sodium channel pore linked to slow inactivation and use dependence. AB - Voltage-gated sodium (Na(+)) channels are a fundamental target for modulating excitability in neuronal and muscle cells. When depolarized, Na(+) channels may gradually enter long-lived, slow-inactivated conformational states, causing a cumulative loss of function. Although the structural motifs that underlie transient, depolarization-induced Na(+) channel conformational states are increasingly recognized, the conformational changes responsible for more sustained forms of inactivation are unresolved. Recent studies have shown that slow inactivation components exhibiting a range of kinetic behavior (from tens of milliseconds to seconds) are modified by mutations in the outer pore P-segments. We examined the state-dependent accessibility of an engineered cysteine in the domain III, P-segment (F1236C; rat skeletal muscle) to methanethiosulfonate ethylammonium (MTSEA) using whole-cell current recordings in HEK 293 cells. F1236C was reactive with MTSEA applied from outside, but not inside the cell, and modification was markedly increased by depolarization. Depolarized F1236C channels exhibited both intermediate (I(M); tau approximately 30 ms) and slower (I(S); tau approximately 2 s) kinetic components of slow inactivation. Trains of brief, 5-ms depolarizations, which did not induce slow inactivation, produced more rapid modification than did longer (100 ms or 6 s) pulse widths, suggesting both the I(M) and I(S) kinetic components inhibit depolarization-induced MTSEA accessibility of the cysteine side chain. Lidocaine inhibited the depolarization dependent sulfhydryl modification induced by sustained (100 ms) depolarizations, but not by brief (5 ms) depolarizations. We conclude that competing forces influence the depolarization-dependent modification of the cysteine side chain: conformational changes associated with brief periods of depolarization enhance accessibility, whereas slow inactivation tends to inhibit the side chain accessibility. The findings suggest that slow Na(+) channel inactivation and use dependent lidocaine action are linked to a structural rearrangement in the outer pore. PMID- 11055995 TI - Mg(2+) modulates voltage-dependent activation in ether-a-go-go potassium channels by binding between transmembrane segments S2 and S3. AB - Extracellular Mg(2+) directly modulates voltage-dependent activation in ether-a go-go (eag) potassium channels, slowing the kinetics of ionic and gating currents (Tang, C.-Y., F. Bezanilla, and D.M. Papazian. 2000. J. Gen. Physiol. 115:319 337). To exert its effect, Mg(2+) presumably binds to a site in or near the eag voltage sensor. We have tested the hypothesis that acidic residues unique to eag family members, located in transmembrane segments S2 and S3, contribute to the Mg(2+)-binding site. Two eag-specific acidic residues and three acidic residues found in the S2 and S3 segments of all voltage-dependent K(+) channels were individually mutated in Drosophila eag, mutant channels were expressed in Xenopus oocytes, and the effect of Mg(2+) on ionic current kinetics was measured using a two electrode voltage clamp. Neutralization of eag-specific residues D278 in S2 and D327 in S3 eliminated Mg(2+)-sensitivity and mimicked the slowing of activation kinetics caused by Mg(2+) binding to the wild-type channel. These results suggest that Mg(2+) modulates activation kinetics in wild-type eag by screening the negatively charged side chains of D278 and D327. Therefore, these residues are likely to coordinate the bound ion. In contrast, neutralization of the widely conserved residues D284 in S2 and D319 in S3 preserved the fast kinetics seen in wild-type eag in the absence of Mg(2+), indicating that D284 and D319 do not mediate the slowing of activation caused by Mg(2+) binding. Mutations at D284 affected the eag gating pathway, shifting the voltage dependence of Mg(2+)-sensitive, rate limiting transitions in the hyperpolarized direction. Another widely conserved residue, D274 in S2, is not required for Mg(2+) sensitivity but is in the vicinity of the binding site. We conclude that Mg(2+) binds in a water-filled pocket between S2 and S3 and thereby modulates voltage dependent gating. The identification of this site constrains the packing of transmembrane segments in the voltage sensor of K(+) channels, and suggests a molecular mechanism by which extracellular cations modulate eag activation kinetics. PMID- 11055996 TI - mu-conotoxin GIIIA interactions with the voltage-gated Na(+) channel predict a clockwise arrangement of the domains. AB - Voltage-gated Na(+) channels underlie the electrical activity of most excitable cells, and these channels are the targets of many antiarrhythmic, anticonvulsant, and local anesthetic drugs. The channel pore is formed by a single polypeptide chain, containing four different, but homologous domains that are thought to arrange themselves circumferentially to form the ion permeation pathway. Although several structural models have been proposed, there has been no agreement concerning whether the four domains are arranged in a clockwise or a counterclockwise pattern around the pore, which is a fundamental question about the tertiary structure of the channel. We have probed the local architecture of the rat adult skeletal muscle Na(+) channel (mu1) outer vestibule and selectivity filter using mu-conotoxin GIIIA (mu-CTX), a neurotoxin of known structure that binds in this region. Interactions between the pore-forming loops from three different domains and four toxin residues were distinguished by mutant cycle analysis. Three of these residues, Gln-14, Hydroxyproline-17 (Hyp-17), and Lys-16 are arranged approximately at right angles to each other in a plane above the critical Arg-13 that binds directly in the ion permeation pathway. Interaction points were identified between Hyp-17 and channel residue Met-1240 of domain III and between Lys-16 and Glu-403 of domain I and Asp-1532 of domain IV. These interactions were estimated to contribute -1.0+/-0.1, -0.9+/-0.3, and -1.4+/-0.1 kcal/mol of coupling energy to the native toxin-channel complex, respectively. mu CTX residues Gln-14 and Arg-1, both on the same side of the toxin molecule, interacted with Thr-759 of domain II. Three analytical approaches to the pattern of interactions predict that the channel domains most probably are arranged in a clockwise configuration around the pore as viewed from the extracellular surface. PMID- 11055997 TI - Functional interactions in Ca(2+) signaling over different time and distance scales. PMID- 11055998 TI - Functional triads consisting of ryanodine receptors, Ca(2+) channels, and Ca(2+) activated K(+) channels in bullfrog sympathetic neurons. Plastic modulation of action potential. AB - Fluorescent ryanodine revealed the distribution of ryanodine receptors in the submembrane cytoplasm (less than a few micrometers) of cultured bullfrog sympathetic ganglion cells. Rises in cytosolic Ca(2+) ([Ca(2+)](i)) elicited by single or repetitive action potentials (APs) propagated at a high speed (150 microm/s) in constant amplitude and rate of rise in the cytoplasm bearing ryanodine receptors, and then in the slower, waning manner in the deeper region. Ryanodine (10 microM), a ryanodine receptor blocker (and/or a half opener), or thapsigargin (1-2 microM), a Ca(2+)-pump blocker, or omega-conotoxin GVIA (omega CgTx, 1 microM), a N-type Ca(2+) channel blocker, blocked the fast propagation, but did not affect the slower spread. Ca(2+) entry thus triggered the regenerative activation of Ca(2+)-induced Ca(2+) release (CICR) in the submembrane region, followed by buffered Ca(2+) diffusion in the deeper cytoplasm. Computer simulation assuming Ca(2+) release in the submembrane region reproduced the Ca(2+) dynamics. Ryanodine or thapsigargin decreased the rate of spike repolarization of an AP to 80%, but not in the presence of iberiotoxin (IbTx, 100 nM), a BK-type Ca(2+)-activated K(+) channel blocker, or omega-CgTx, both of which decreased the rate to 50%. The spike repolarization rate and the amplitude of a single AP-induced rise in [Ca(2+)](i) gradually decreased to a plateau during repetition of APs at 50 Hz, but reduced less in the presence of ryanodine or thapsigargin. The amplitude of each of the [Ca(2+)](i) rise correlated well with the reduction in the IbTx-sensitive component of spike repolarization. The apamin-sensitive SK-type Ca(2+)-activated K(+) current, underlying the afterhyperpolarization of APs, increased during repetitive APs, decayed faster than the accompanying rise in [Ca(2+)](i), and was suppressed by CICR blockers. Thus, ryanodine receptors form a functional triad with N-type Ca(2+) channels and BK channels, and a loose coupling with SK channels in bullfrog sympathetic neurons, plastically modulating AP. PMID- 11055999 TI - Opening and closing of KCNKO potassium leak channels is tightly regulated. AB - Potassium-selective leak channels control neuromuscular function through effects on membrane excitability. Nonetheless, their existence as independent molecular entities was established only recently with the cloning of KCNKO from Drosophila melanogaster. Here, the operating mechanism of these 2 P domain leak channels is delineated. Single KCNKO channels switch between two long-lived states (one open and one closed) in a tenaciously regulated fashion. Activation can increase the open probability to approximately 1, and inhibition can reduce it to approximately 0.05. Gating is dictated by a 700-residue carboxy-terminal tail that controls the closed state dwell time but does not form a channel gate; its deletion (to produce a 300-residue subunit with two P domains and four transmembrane segments) yields unregulated leak channels that enter, but do not maintain, the closed state. The tail integrates simultaneous input from multiple regulatory pathways acting via protein kinases C, A, and G. PMID- 11056001 TI - Overexpression of protectin (CD59) down-modulates the susceptibility of human melanoma cells to homologous complement. AB - The clinical efficacy of therapeutic complement (C)-activating monoclonal antibodies (mAb) to melanoma-associated antigens can be impaired by the levels of expression of C-inhibitory molecules on neoplastic cells. Protectin (CD59) is a glycosylphosphatidylinositol (GPI)-anchored cell membrane glycoprotein, acting as terminal regulator of C cascade, which is heterogeneously expressed in melanomas and represents the main restriction factor of C-mediated lysis of melanoma cells. Thus, we investigated whether the overexpression of CD59 could influence the constitutive susceptibility of distinct melanoma cells to homologous C. Infection of CD59-positive Mel 100 and 70-W melanoma cells by a retroviral vector carrying the CD59 cDNA, significantly (P < 0.05) upregulated their constitutive expression of CD59, whereas it did not affect that of additional C-regulatory molecules. Transduced CD59 was entirely GPI-anchored and showed a molecular weight identical to native CD59. Additionally, higher amounts of soluble CD59 were detected in the conditioned media of CD59-transduced melanoma cells compared with parental cells. CD59-transduced melanoma cells, sensitized by the anti-GD3 disialoganglioside mAb R24, were significantly (P < 0.05) less susceptible to homologous C-lysis than were parental cells; this effect was fully reverted by the masking of CD59 with F(ab')(2) fragments of the anti-CD59 mAb YTH53.1. These results provide conclusive evidence demonstrating that absolute levels of CD59 expression regulate the susceptibility to homologous C of specific melanoma cells, and suggest an additional explanation for the poor clinical results obtained with C activating mAb in the clinical setting. PMID- 11056002 TI - Noradrenaline induces brown adipocytes cell growth via beta-receptors by a mechanism dependent on ERKs but independent of cAMP and PKA. AB - It has been well established that the key role of noradrenaline is the induction of uncoupling-protein-1 (UCP-1) expression, the unique marker of brown adipocytes. However, its implication on proliferation and the pathways involved are not as well characterized. By using rat fetal brown adipocytes as a model, we show that, although noradrenaline activates extracellular regulated kinases (ERKs) through beta-, alpha1-, and alpha2-receptors, only beta-receptors mediate cell growth by a mechanism that requires ERKs activation but is independent of cyclic-adenosine-monophosphate/protein kinase A (cAMP/PKA). Conversely, the cAMP/PKA cascade mediates noradrenaline-induced UCP-1 expression, whereas ERKs pathway attenuates thermogenic differentiation. On the other hand, alpha1- and alpha2-receptors have an antiproliferative effect that is enhanced by ERK inhibition. PMID- 11056003 TI - Interferon-gamma sensitizes colonic epithelial cell lines to physiological and therapeutic inducers of colonocyte apoptosis. AB - Homeostasis in the colonic epithelium is achieved by a continuous cycle of proliferation and apoptosis, in which imbalances are associated with disease. Inflammatory bowel disease (IBD) and colon cancer are associated with either excessive or insufficient apoptosis of colonic epithelial cells, respectively. By using two colonic epithelial cell lines, HT29 and SW620, we investigated how the epithelial cell's sensitivity to apoptosis was regulated by the proinflammatory cytokine interferon-gamma (IFN-gamma). We found that IFN-gamma sensitized HT29 cells, and to a lesser extent SW620, to diverse inducers of apoptosis of physiologic or therapeutic relevance to the colon. These apoptosis inducers included Fas (CD95/APO-1) ligand (FasL), short-chain fatty acids, and chemotherapeutic drugs. The extent of IFN-gamma-mediated apoptosis sensitization in these two cell lines correlated well with the degree of IFN-gamma-mediated upregulation of the proapoptotic protease caspase-1. Although IFN-gamma alone effectively sensitized HT29 cells to apoptosis, inclusion of the protein synthesis inhibitor cyclohexamide (CHX) during apoptotic challenge was necessary for maximal sensitization of SW620. The requirement of CHX to sensitize SW620 cells to apoptosis implies a need to inhibit translation of antiapoptotic proteins absent from HT29. In particular, the antiapoptotic protein Bcl-2 was strongly expressed in SW620 cells but absent from HT29. Our results indicate that IFN-gamma increases the sensitivity of colonic epithelial cells to diverse apoptotic stimuli in concert, via upregulation of caspase-1. Our findings implicate caspase-1 and Bcl-2 as important central points of control determining the general sensitivity of colonic epithelial cells to apoptosis. PMID- 11056004 TI - Expression and localization of rab escort protein isoforms in parotid acinar cells from rat. AB - Rab proteins are geranylgeranylated on their carboxyl terminal cysteine motifs by geranylgeranyltransferase II (GGTase). Rab escort protein (REP) is required to present Rab proteins to GGTase. REP may remain bound to newly isoprenylated Rab proteins and present them to their target membrane. Other studies have shown that Rab proteins cycle between the membrane and cytosolic compartments and that cytosolic Rab proteins are complexed with rab-GDI. In the present study, we examined the expression and localization of REP isoforms in parotid acinar cells. Although both REP isoforms, REP-1 and REP-2, were detected in parotid cytosol, REP-2 was the predominant isoform. Subcellular fractionation revealed that approximately 42% of cellular REP-2 is membrane-associated. REP-2 was partially removed from parotid membranes with 1 M NaCl or Na(2)CO(3), indicating that REP-2 is a peripheral membrane protein. Membrane-associated REP-2 did not colocalize with Rab3D on secretory granule membranes. However, density gradient centrifugation revealed that membrane-associated REP-2 and Rab3D colocalize on low- and high-density membrane fractions in parotid acinar cells. Isoproterenol, an agent which induces amylase release from parotid glands, caused a shift in both REP-2 and Rab3D to less dense membrane fractions. When acinar cell cytosol was fractionated by gel filtration chromatography, Rab3D eluted exclusively with REP, not rab-GDI. In contrast, Rab1B and Rab5 eluted with both REP and Rab-GDI. Colocalization of Rab3D and REP-2 on acinar cell membranes suggests that REP-2 plays a role in delivering Rab3D to parotid membranes and may regulate guanine nucleotide binding to membrane-associated Rab3D. In addition, unlike other Rab proteins, cytosolic Rab3D appears to associate exclusively with REP, not rab-GDI in parotid acinar cells. PMID- 11056005 TI - Pervanadate induces the hyperphosphorylation but not the activation of human heat shock factor 1. AB - In this study, we evaluated the effects of pervanadate, a tyrosine phosphatase inhibitor, on the regulation and function of heat-shock factor 1 (HSF1) in HeLa cells. We showed that 50-100 microM pervanadate induced the hyperphosphorylation of the latent HSF1, as demonstrated by a retarded mobility of the HSF1 protein in SDS-polyacrylamide gel electrophoresis and as supported by the reversal of this mobility shift upon treatment of the cell extract with acid phosphatase. Pervanadate by itself had no effect on the monomeric stoichiometry and DNA binding activity of HSF1. Upon heat shock, the pervanadate-induced hyperphosphorylated HSF1 formed DNA-binding trimers and translocated into the nuclear compartment. At high concentration (approximately 500 microM), pervanadate also induced the tyrosine phosphorylation of many cellular proteins and blunted the heat-induced transcription of hsp 70. N-acetyl cysteine inhibited these effects of pervanadate, suggesting a redox-based mechanism for its activity. Analysis of the activation of mitogen-activated protein kinases (MAPKs) using antibodies specific for the phospho-form (activated) of the kinases in Western blot showed that pervanadate activated extracellular signal-regulated kinase (ERK1/2), c-Jun-N-terminal kinase 1/2 (JNK1/2), and p-38 kinase. Pharmacological inhibitors of the ERK1/2 kinase pathway or the p38 kinase had little or no effect on the pervanadate-induced hyperphosphorylation of HSF1. Our results show that hyperphosphorylation of hHSF1 can occur prior to and independent of other events involved in the activation of hHSF1. The possibility that activation of the MAPK signaling cascade, notably JNK, may contribute to the hyperphosphorylation of human HSF1 (hHSF1) is discussed. PMID- 11056006 TI - Localization of BAI-associated protein1/membrane-associated guanylate kinase-1 at adherens junctions in normal rat kidney cells: polarized targeting mediated by the carboxyl-terminal PDZ domains. AB - Brain-specific angiogenesis inhibitor (BAI)-associated protein (BAP)1 (also called membrane-associated guanylate kinase [MAGI]-1) is composed of six PSD 95/Dlg-A/ZO-1 (PDZ) domains, two WW domains, and one guanylate kinase (GK) domain. We previously reported that BAP1 is localized at tight junctions in Madine Darby canine kidney (MDCK) cells and intestinal epithelial cells. Here, we have determined the localization of BAP1 in normal rat kidney (NRK) cells that do not form tight junctions. BAP1 was colocalized with E-cadherin along the lateral membrane, suggesting its localization at adherens junctions. Green fluorescent protein (GFP)-BAP1 was distributed in the cytosol in separate NRK cells, and accumulated to the cell-cell contacts when NRK cells have contact with each other. The GFP-BAP1 mutant containing either the first PDZ and GK domains or the WW and second PDZ domains was localized in the cytosol and the nucleus. The GFP BAP1 mutant containing the second to fourth PDZ domains was distributed in the cytosol. The construct containing the fifth and sixth PDZ domains was localized at the cell-cell contacts along the lateral membrane and slightly in the nucleus, whereas the construct lacking the fifth and sixth PDZ domains was localized in the cytosol and in the nucleus. BAP1 was tyrosine-phosphorylated in vivo, but the tyrosine phosphorylation of BAP1 was not correlated with its localization. These results suggest that the signal in the carboxyl-terminal PDZ domains functions dominantly in vivo to target BAP1 to the lateral membrane, although potential nuclear localization signals exist in the N-terminal region of BAP1. PMID- 11056007 TI - Rat testicular myotubularin, a protein tyrosine phosphatase expressed by Sertoli and germ cells, is a potential marker for studying cell-cell interactions in the rat testis. AB - The full-length cDNA encoding the entire open reading frame (ORF) of rat myotubularin (rMTM) was isolated from a rat testis expression library by PCR. Among the three approximately 2.9-kb cDNAs that were sequenced, one clone was different from the other two clones. It contained seven extra amino acids of FVVLNLQ; this short stretch of extra sequence was found between Gln(421) and Phe(422) within the SET (Suvar3-9, Enhancer-of-zeste, Trithorax) interacting domain (SID) of rMTM. The rMTM ORF had 1,713 bp encoding for a 571 amino acid polypeptide and a calculated molecular weight of 65.8 kDa. A comparison between its deduced amino acid sequence and the GenBank database using BLAST revealed a 53.1% identity with human myotubularin protein (hMTM1), which is a member of the protein tyrosine phosphatase (PTP) family associated with X-linked myotubular myopathy. A 22 amino acid peptide NH(2)-TKVNERYELCDTYPALLAVPAN was synthesized based on the deduced amino acid sequence of rMTM and used for antibody production. By using immunoblot analysis, a 66-kDa protein was indeed detected in both Sertoli and germ-cell cytosols. rMTM mRNA was found in various tissues but was predominantly expressed in the testis, ovary, and skeletal muscle. Sertoli cell rMTM expression was stimulated by germ cells and enhanced when inter-Sertoli junctions were being assembled in vitro. A drastic reduction in testicular rMTM steady-state mRNA level correlated with the depletion of germ cells from the testis in vivo following either glycerol or lonidamine treatment. These results indicate that rMTM is a rat homologue of hMTM1 that may be a useful marker in monitoring the events of cell-cell interactions in the testis. PMID- 11056008 TI - Growth inhibition and protein tyrosine phosphorylation in MCF 7 breast cancer cells by a novel K vitamin. AB - We have now found that the most potent, Cpd 5 [2-(2-mercaptoethanol)-3-methyl-1, 4-napthoquinone], inhibits growth of doxorubicin-resistant and doxorubicin sensitive breast cancer cells (MCF 7r and MCF 7w) in culture. Growth inhibition by Cpd 5 was antagonized by the thiol antioxidants glutathione and cysteine, but not by catalase or superoxide dismutase, suggesting that growth inhibition is probably via conjugation of cellular thiols. In support of this, we found that Cpd 5 inhibited the activity of thiol containing cellular protein tyrosine phosphatase (PTP) enzyme, with consequent induction of various tyrosine phosphoproteins, but not serine or tyrosine phosphoproteins. The tyrosine phosphorylation was also inhibited by exogenous glutathione or cysteine and could be enhanced by depletion of cellular glutathione by BSO. This effect of Cpd 5 on protein tyrosine phosphorylation was highly selective, however. Tyrosine phosphorylation of EGF-R, Erb-B2, and ERK1/2 was increased, but not that of Insulin-R or JNK. ERK1/2 tyrosine phosphorylation and growth inhibition increased with increasing concentrations of Cpd 5. Furthermore, suppression of Cpd 5 mediated ERK1/2 phosphorylation by an ERK-kinase inhibitor antagonized growth inhibition. These results suggest a strong correlation between ERK1/2 phosphorylation by Cpd 5 and growth inhibition. This novel K-vitamin analog thus inhibits MCF 7 cell growth and induces selective protein tyrosine phosphorylation. PMID- 11056009 TI - Downregulation of platelet-activating factor responsiveness during maturation of human dendritic cells. AB - Dendritic cells (DCs) are specialized antigen-presenting cells characterized by their ability to migrate into target sites, process antigens, and activate naive T-cells. Biological activities of platelet-activating factor (PAF) and the cytokine macrophage inflammatory protein-3beta (MIP-3beta) as well as the mRNA expression of their receptors were characterized in human DCs during lipopolysaccharide (LPS)-promoted maturation. Platelet-activating factor induced calcium transients, migration-associated actin polymerization response, and chemotaxis in immature human dendritic cells differentiated in vitro from monocytes with interleukin-4 and granulocyte macrophage colony stimulating factor. In addition, RT-PCR experiments indicated mRNA expression of the PAF receptor in these immature DCs. Cell studies and mRNA analyses further revealed that immature DCs neither respond to MIP-3beta nor express its specific receptor, CCR7. Induction of cell differentiation by LPS led to the loss of the mRNA expression of the PAF receptor, accompanied by decreasing intracellular calcium release, actin polymerization, and migration after stimulation with PAF. In contrast, LPS treatment induced increasing responsiveness toward MIP-3beta and mRNA expression of CCR7. Comparable data regarding mRNA expression of PAF receptor and PAF responsiveness were also obtained with another maturation protocol using TNFalpha instead of LPS. The direct comparison between the two different protocols showed a slower decrease of PAF responsiveness induced by TNFalpha than by LPS. These results show the loss of PAF responsiveness associated with downregulation of PAF receptor mRNA expression during LPS- and TNFalpha-induced maturation in human DCs. Therefore, these findings point to a functional relevance of PAF in recruiting immature DCs, whereas MIP-3beta might regulate the migration of DCs at a later stage of maturation. PMID- 11056010 TI - Estrogen-induced resistance to osteoblast apoptosis is associated with increased hsp27 expression. AB - Estrogen has been shown to protect osteoblastic cells from apoptosis. Similarly, estrogen treatment preceding heat shock elevates heat shock protein 27 (hsp27) expression and increases thermoresistance in the murine estrogen receptor transformed SMER14 osteoblastic cell line. Forced expression of hsp27 expression in other cell lines limits apoptosis. The purpose of this study was to examine the effects of estrogen on staurosporine-induced apoptosis in the context of hsp27 expression. Cell viability was measured by the MTT assay. Early apoptotic events were examined by fluorescent microscopy by using FITC-conjugated Annexin V staining. TUNEL labeling was used to compare the number of apoptotic nuclei following staurosporine treatment of estrogen pretreated or untreated cells. Estrogen treatment increased SMER14 cell viability, but not ROS17/2.8 cell viability, in the presence of staurosporine. Estrogen treatment also reduced annexin V staining and DNA fragmentation. Similar treatment increased SMER14 cell hsp27 levels. The concurrent reduction in induced apoptosis suggests a possible estrogenic mechanism for increasing and/or maintaining the number of viable osteoblasts in bone. PMID- 11056011 TI - Protein kinase-A-mediated secretion of mucin from human colonic epithelial cells. AB - Both Ca(2+)- and cAMP-mediated second messenger cascades acutely regulate mucin secretion from human colonic epithelial cells. To better understand the cAMP dependent regulation of mucin secretion we have characterized the complement of cAMP-dependent protein kinase (PKA) isoforms in mucus-secreting T84 cells, and determined which of these isoforms is responsible for agonist-stimulated mucin secretion. Our results show the presence of both type I and type II PKA in cells that also contain large mucin granules. Forskolin caused a rapid and sustained increase in PKA activity that reached a maximum 5-10 min following its addition. Secretion of mucin was detected 15 min following exposure to forskolin, and continued to increase for a further 15 min before reaching a plateau. Mucin secretion was also measured in the presence of combinations of site-selective cAMP analog pairs, which preferentially activate either type I or type II PKA. Similar levels of mucin secretion were observed for both type I and type II PKA selective analog pairs. Subsequent addition of forskolin was unable to further increase mucin secretion. Thus, activation of either type I or type II PKA is able to maximally stimulate secretion of mucins from T84 human colonic epithelial cells. PMID- 11056012 TI - Transforming growth factor-beta is an autocrine mitogen for a novel androgen responsive murine prostatic smooth muscle cell line, PSMC1. AB - A prostatic smooth muscle cell line (PSMC1) was established from the dorsolateral prostate of p53 null mice. The cell line is nontumorigenic when inoculated subcutaneously, under the renal capsule or intraprostatically in syngeneic mice. These cells express alpha-smooth muscle actin (alpha-SMA), indicating their smooth muscle origin, and TGF-beta significantly enhances expression of alpha SMA. The cells express both androgen receptor (AR) mRNA and protein, and respond mitogenically to physiological concentrations of androgens. PSMC1 cells produce significant amounts of TGF-beta, which stimulates growth by an autocrine mechanism. Dihydrotestosterone (DHT) increases proliferation of PSMC1 cells by promoting TGF-beta secretion. Considering the significant inhibitory effect of TGF-beta on prostatic epithelial cells and its stimulatory effect on the PSMC1 cells, we postulate that TGF-beta produced by prostatic smooth muscle cells may have a paracrine effect on the prostatic epithelium. We also postulate that TGF beta may be involved in the etiology of benign prostatic hyperplasia (BPH) by stimulating excessive stromal proliferation. Line PSMC1 is the first reported androgen-responsive murine smooth muscle cell line. It will be useful for in vivo and in vitro experiments to study the mechanisms of androgen action on prostatic stroma and for delineating the interactions that occur between prostatic smooth muscle and epithelium that may lead to prostatic diseases such as BPH. PMID- 11056013 TI - Jagged2 induces cell cycling in confluent fibroblasts susceptible to density dependent inhibition of cell division. AB - Jagged2 is a member of the DSL (Delta-Serrate-Lag-2) -ligand family of transmembrane proteins that signal through the Notch receptors. In many cases of human acute lymphoblastic T-cell leukemias, chromosomal translocations fuse a part of the Notch-1 gene to the T-cell receptor-beta locus (Ellison et al., 1991, Cell 66:649-661). The truncated Notch-1 allele encodes an aberrant protein that lacks most of the extracellular domain and is constitutively activated (Pear et al., 1996, J Exp Med 183:2283-2291). A similarly truncated version of Notch-1 was capable of transforming primary baby rat kidney cells in cooperation with the E1A oncogene of adenovirus (Capobianco et al., 1997, Mol Cell Bio 17:6265-6273). The transformed cells grew to a high population density in culture and were tumorigenic in vivo. It was unclear what roles Notch signaling played in neoplastic transformation. In this report, we demonstrate that sustained activation of the Jagged2/Notch signal transduction pathway induced continuous cell cycling in confluent rabbit-skin fibroblasts sensitive to density-dependent inhibition of cell division. The ability to overcome density-dependent inhibition of cell division correlated with elevated cyclin-dependent kinase-2 (CDK2) activity and a lower level of induction of the CDK inhibitor p27 in the target cells. Similar cell-cycle effect was seen when a truncated mouse Notch-1 construct with constitutive activity was expressed. Taken together, our findings indicate that sustained activation of the Jagged2/Notch signal transduction pathway can overcome density-dependent inhibition of cell division and therefore may contribute to neoplastic transformation. PMID- 11056014 TI - Differences in the mechanism for high- versus moderate-density fibroblast populated collagen lattice contraction. AB - The free-floating fibroblast-populated collagen lattice (FPCL) model introduced by Bell contains 0.5 x 10(5) cell/ml and here is defined as a moderate-density FPCL (MD-FPCL). One modification of the model is to increase the cell density by a factor of 10, where 5 x 10(5) cells/ml defines a high-density FPCL (HD-FPCL). The initial detection of HD-FPCL contraction is 2 h, whereas MD-FPCL is later, 6 h. A contracted HD-FPCL has a doughnut-like appearance, due to the high density of cells accumulating at the periphery. A contracted MD-FPCL is a flattened disc. The compacted collagen of MD-FPCL lattice exhibits a strong birefringence pattern due to organized collagen fiber bundles. In contracted HD-FPCL, a minimal birefringence develops, indicating minimal organization of collagen fiber bundles. MD-FPCL contraction was reduced with less than 10% serum; the disruption of microtubules, uncoupling of gap junctions, inhibition of tyrosine kinases, and addition of a blocking antibody to alpha2beta1 collagen integrin. Making HD-FPCL with only 1% serum or including the inhibitory agents had only minimal affect on lattice contraction. On the other hand, platelet-derived growth factor stimulated HD-FPCL contraction but had no influence on MD-FPCL contraction. It is suggested that the mechanism for HD-FPCL contraction is limited to the process of cells spreading. HD-FPCL contraction is independent of collagen organization, microtubules, gap junctions, alpha2beta1 integrin, and tyrosine phosphorylation. MD-FPCL contraction involves collagen organization and is optimized by the involvement of microtubules, gap junctions, alpha2beta1 integrin, and tyrosine phosphorylation. When studying cell physiology in a collagen matrix, cell-density influences need to be considered. PMID- 11056015 TI - The transport mechanism of metallothionein is different from that of classical NLS-bearing protein. AB - A nuclear localization signal (NLS) has been detected in several nuclear proteins. Classical NLS-mediated nuclear pore targeting is performed by using the cytosolic factors, importin alpha and importin beta, whereas nuclear translocation requires the small GTPase, Ran. In the present study, we demonstrated that nuclear localization of metallothionein (MT) differs from that of classical NLS-mediated substrates. In digitonin-permeabilized BALB/c3T3 cells, biotinylated MT was localized in the nucleus in the presence of ATP and erythrocyte cytosol in the same manner as for SV40 large T NLS-conjugated allophycocyanin (APC-NLS). Under ATP-free conditions, nuclear rim-binding was observed in both transport substrates. Rim-binding of labeled MT was competitively inhibited by the addition of an excess amount of unlabeled MT. Different elution profiles were observed for the localization-promoting activities of MT in the cytosol compared to those of APC-NLS. Furthermore, nuclear localization of MT was determined to be a wheat germ agglutinin insensitive, GTPgammaS-sensitive, and anti-Ran antibody-sensitive process. Green fluorescent protein-metallothionein (GFP-MT) fusion protein was also localized in the nucleus in the stable transformant of CHL-IU cells. These results strongly suggest that the targeting by MT of the nuclear pore is mediated by cytosolic factor(s) other than importins and that MT requires Ran for its nuclear localization. PMID- 11056016 TI - Motogenic effect of recombinant HGF on airway epithelial cells during the in vitro wound repair of the respiratory epithelium. AB - Cell migration is the earliest mechanism involved in the wound repair process of the respiratory epithelium and could be potentially enhanced by growth factors. In the present work, we investigated the localisation of the hepatocyte growth factor (HGF) receptor (c-Met) during wound repair and evaluated the effect of recombinant HGF (rHGF) on cell migration by using an in vitro model of airway epithelial wound repair. By using immunohistochemical methods, we observed that the immunoreactivity of the c-Met proto-oncogene was increased in epithelial cells engaged in the process of tissue repair. The incubation of wounded cultures with increasing concentrations of rHGF (0.2, 2, 20, and 200 ng/ml) induced a significant (P < 0.02) dose-dependent effect on the wound repair index, with a maximum effect produced at 20 ng/ml (+31.3%). The cell migration speed reached 50.2 micrometer/h at this concentration, compared to 20.4 micrometer/h in the absence of rHGF. No significant effect on cell proliferation was observed in the repairing area in the presence of rHGF. These results suggest that rHGF is able to improve the wound repair process of the airway epithelium by increasing cell migration. PMID- 11056017 TI - Calcium influx induced by activation of tyrosine kinase receptors in cultured bovine aortic endothelial cells. AB - We studied the ionic currents activated by basic fibroblast growth factor (bFGF) and insulin-like growth factor-I (IGF-I) in cultured bovine aortic endothelial cells (BAE-1) by using patch-clamp and single-cell fluorimetric calcium measurements. In whole-cell, voltage-clamp experiments at V(h) = -50 mV, the addition of either bFGF (20 ng/ml) or IGF-I (50 ng/ml) induced an inward current with similar amplitude, time course, and permeation properties. The response was dependent on receptor occupancy and showed a desensitisation in the continued presence of the factors. Ionic substitutions in whole-cell experiments indicated that the current barely discriminated among Na(+), Ca(+), and K(+) ions. Accordingly, stimulation with bFGF or IGF-I induced a dose-dependent [Ca(2+)](i) elevation completely due to entry from the extracellular medium, whereas no detectable release from internal stores was observed. Calcium influx was dependent on protein tyrosine kinase (PTK) activity; it was significantly inhibited by treatment with genistein or tyrphostin 47, two PTK inhibitors, and not affected by inactive analogues, daidzein, and tyrphostin 1. Moreover, addition of 200 microM Na(3)VO(4), an inhibitor of protein tyrosine phosphatase (PTP) activity, evoked the responses to the factors both in patch-clamp and in fluorimetric measurements. Cell-attached recordings using 100 mM CaCl(2) in the pipette showed that bFGF and IGF-I activate calcium-permeable channels with similar properties. These results provide evidence for a calcium influx induced by two factors that bind to tyrosine kinase receptors (RTK) in endothelial cells. PMID- 11056018 TI - Roles of zinc fingers and other regions of the transcription factor human MTF-1 in zinc-regulated DNA binding. AB - Mammalian metallothionein genes are transcriptionally regulated by heavy metals through cis-acting metal responsive elements (MREs). The MRE-binding transcription factor-1 (MTF-1), a protein containing six C(2)H(2)-type Zn fingers, is essential for MRE-mediated transcriptional activation. DNA binding of MTF-1 is known to be stimulated by Zn in vitro, but the binding was also largely influenced by redox conditions, suggesting that redox signals could modulate MTF 1 activity. To locate the functional domain required for Zn regulation, several deletion mutants of human MTF-1b, a newly cloned transcriptionally active MTF-1 variant, were characterized. This analysis showed that the N-terminal region and Zn-finger domain play roles in metal response. Functional roles of individual Zn fingers were estimated by co-transfection assays by using an MRE-driven reporter gene and vectors that express MTF-1b mutants each carrying one defective finger. Mutations in the N-terminal four fingers dramatically reduced the transcriptional activity, and at least for three of them the transcriptional defect was due to reduced DNA binding. These results indicate that the six Zn fingers are not functionally equivalent, probably sharing distinct roles such as direct DNA recognition and regulatory functions. PMID- 11056019 TI - Investigation of the substrate spectrum of the human mismatch-specific DNA N glycosylase MED1 (MBD4): fundamental role of the catalytic domain. AB - The human DNA repair protein MED1 (also known as MBD4) was isolated as an interactor of the mismatch repair protein MLH1 in a yeast two-hybrid screening. MED1 has a tripartite structure with an N-terminal 5-methylcytosine binding domain (MBD), a central region, and a C-terminal catalytic domain with homology to bacterial DNA damage-specific glycosylases/lyases. Indeed, MED1 acts as a mismatch-specific DNA N-glycosylase active on thymine, uracil, and 5-fluorouracil paired with guanine. The glycosylase activity of MED1 preferentially targets G:T mismatches in the context of CpG sites; this indicates that MED1 is involved in the repair of deaminated 5-methylcytosine. Interestingly, frameshift mutations of the MED1 gene have been reported in human colorectal, endometrial, and pancreatic cancers. For its putative role in maintaining genomic fidelity at CpG sites, it is important to characterize the biochemical properties and the substrate spectrum of MED1. Here we show that MED1 works under a wide range of temperature and pH, and has a limited optimum range of ionic strength. MED1 has a weak glycosylase activity on the mutagenic adduct 3,N(4)-ethenocytosine, a metabolite of vinyl chloride and ethyl carbamate. The differences in glycosylase activity on G:U and G:T substrates are not related to differences in substrate binding and likely result from intrinsic differences in the chemical step. Finally, the isolated catalytic domain of MED1 retains the preference for G:T and G:U substrates in the context of methylated or unmethylated CpG sites. This suggests that the catalytic domain is fundamental, and the 5-methylcytosine binding domain dispensable, in determining the substrate spectrum of MED1. PMID- 11056020 TI - International conference on basic and clinical aspects of cell-cycle control. AB - Scientists of numerous medical and life science disciplines met in Siena, Italy to discuss the latest proceedings in basic and clinical research. General models of interconnected linear and back-feeding cell-cycle control pathways provide a basis for applied molecular research. Cell-cycle determining factors essential for the control of cellular homeostasis either become markers to determine characteristics of a disease and/or become therapeutic targets. Apart from animal and tissue culture models, molecular theories finally have to stand proof in clinical application and evaluation. Therefore, the clinical feedback to the basic scientist's bench is essential for necessary adjustments of their models to improve future approaches to research challenges. A select group of speakers provided the audience with such an interdisciplinary dialogue at the first International Conference on Basic and Clinical Aspects of Cell-Cycle Control from May 29 to 31, 2000 in Siena, Italy. PMID- 11056021 TI - Three-dimensional power Doppler sonography in screening for carotid artery disease. AB - PURPOSE: Color Doppler sonography has gained considerable recognition as a noninvasive method to detect carotid artery disease and to assess the degree of carotid artery stenosis. However, results are highly operator-dependent and cannot be presented as survey images. The purpose of this study was to evaluate real-time 3-dimensional (3D) power Doppler sonography as a method for screening for atherosclerosis in the carotid arteries. METHODS: We prospectively screened 75 patients for carotid artery disease using both conventional color Doppler sonography and 3D power Doppler sonography, and the results from the 2 modalities were compared. A total of 150 common carotid arteries, 150 internal carotid arteries, and 150 external carotid arteries were examined utilizing a 7.5-MHz linear-array transducer combined with tissue harmonic imaging. RESULTS: Color Doppler sonography detected 297 normal or atherosclerotic arteries without stenosis, 57 arteries with mild (1-49%) stenosis, 41 with moderate (50-69%) stenosis, 32 with severe (70-99%) stenosis, and 9 with occlusions. The degree of stenosis determined by color Doppler sonography correlated with that determined by 3D power Doppler sonography (r = 0.982-0.998). Moreover, there was a good correlation between the measurements for both the length of the lesion and its distance from the bulb as determined by the 3D volume surveys and by color Doppler sonography (r = 0.986). The interobserver variability rate was 3.7% +/- 0.5%. Generally, the acquisition and reconstruction of the 3D data took less than 5 minutes. CONCLUSIONS: 3D power Doppler sonography is easy to perform and is an accurate method in screening for atherosclerotic lesions of the carotid arteries. Moreover, it provides excellent 3D volume surveys that may be helpful in the planning of surgical treatment. PMID- 11056022 TI - Evaluation of posterior cerebral artery blood flow with transcranial Doppler sonography: value and risk of common carotid artery compression. AB - PURPOSE: Investigations of the posterior cerebral arteries (PCA) by transcranial Doppler sonography (TCD) may be less reliable than investigations of the anterior part of the circle of Willis. Nevertheless, a true PCA may be identified by manual compression of the proximal common carotid artery (CCA) during TCD. Therefore, we used CCA compression in clinically indicated TCD studies and assessed retrospectively its risks and prospectively its benefits for PCA evaluations. METHODS: Using the transtemporal approach, we prospectively assessed flow velocities in posteriorly located blood vessels in 180 consecutive patients before and during CCA compression. The complications of CCA compression were retrospectively reviewed in all 3,383 clinical TCD investigations performed over an 8-year period. RESULTS: Decreased flow velocities during ipsilateral CCA compression occurred in 17% of patients. A PCA-like vessel with perfusion from the carotid artery or PCA supply from the carotid circulation was unmasked. Mixed distal PCA support by the posterior communicating artery and proximal PCA could not be shown by TCD. Transient cerebral symptoms occurred in less than 0.4% of the 3,383 retrospectively reviewed TCD investigations; no other adverse effects were seen. CONCLUSIONS: TCD without CCA compression may lead to false identification of the PCA. Since transient cerebral symptoms during CCA compression are rare, CCA compression can be used when a clinical TCD investigation of intracranial collateral blood flow compensation is indicated or when the identification of a cerebral artery is uncertain. PMID- 11056023 TI - Correlation of MRI liver volume and doppler sonographic portal hemodynamics with histologic findings in patients with chronic hepatitis C. AB - PURPOSE: The purpose of this study was to correlate portal hemodynamics on sonography and liver volume on MRI with histologic findings in asymptomatic patients with chronic hepatitis C. METHODS: Portal blood flow in the left and right portal branches in 20 healthy volunteers and in 26 patients was measured using Doppler sonography during both fasting and postprandial states. Total liver and right-and left-lobe volumes were determined using MRI. The ratio between portal blood flow and liver volume determined the "portal flow index" of the right and left lobes. RESULTS: We observed a statistically significant difference (p < 0.01) between the volunteers and patients in the mean left-lobe volume (352 +/- 81 cm(3) versus 544 +/- 159 cm(3)) and in the mean left portal flow index (1.1 +/- 0.2 ml/minute/cm(3) versus 0.69 +/- 0.2 ml/minute/cm(3)) as measured before the subjects ate. After a meal, the portal blood-flow volume in the right lobe was similar in the 2 groups but in the left lobe was significantly lower in the patients (p = 0.0009). The left postprandial portal flow index was inversely correlated with the grade of liver fibrosis (r = 0.533). CONCLUSIONS: The left lobe volume (positive predictive value, 83%; negative predictive value, 72%) and left postprandial portal flow index (positive predictive value, 86%; negative predictive value, 88%) are sensitive indicators of chronic hepatitis. The left postprandial portal flow index may be a useful test for differentiating patients with minimal or no fibrosis from patients with mild to severe fibrosis. PMID- 11056024 TI - Sonographically guided percutaneous treatment of hepatic hydatid cysts: long-term results. AB - PURPOSE: The purpose of this study was to evaluate the safety and efficacy of sonographically guided percutaneous drainage and irrigation of hepatic hydatid cysts. METHODS: Sixty-one patients with 84 hepatic hydatid cysts were treated using the puncture, aspiration, injection, and reaspiration (PAIR) technique under sonographic guidance. Patients with cysts larger than 6 cm in diameter underwent PAIR followed by percutaneous drainage (PAIR-PD). The cysts were sterilized by the injection of 1 of 2 scolicidal agents, 20% hypertonic saline solution (38 patients) or 0.5% silver nitrate (23 patients). All patients underwent follow-up examinations for 1 month-6 years after aspiration. Clinical and radiologic examinations and laboratory analyses were performed every month for the first 6 months and then at 3-month intervals. RESULTS: Serial sonographic examinations revealed a heterogeneous echo pattern in 78 cysts (93%); a progressive decrease in diameter in 76 cysts (90%); calcification of the cyst wall, cystic contents, or both in 10 cysts (12%); and complete disappearance of 1 cyst (1%) in a patient who had been monitored for over 6 years. Five patients developed urticaria, and 6 developed fever. One patient developed a biliary fistula after the first aspiration attempt. Two patients developed infection of the cyst cavity after PAIR-PD and were successfully treated with oral antibiotics. An anaphylactic reaction developed in 2 patients and was successfully treated with antiallergenic medication. No recurrence of hydatid disease after PAIR or PAIR-PD was observed in any patient over the follow-up period of 72 months (mean, 26 +/- 27 months). CONCLUSIONS: Percutaneous drainage of hydatid cysts is a safe, effective, and reliable treatment. Antiallergenic medication is required before PAIR or PAIR-PD. Both sclerosing agents, hypertonic saline and silver nitrate solutions, gave excellent results. PMID- 11056025 TI - Technical factors influencing sonographic visualization of fetal echogenic intracardiac foci. AB - PURPOSE: A fetal echogenic intracardiac focus (EIF) is most commonly a normal variant in a normal fetus, but owing to reports of an increased risk of aneuploidy with EIFs, the finding causes concern when noted on routine obstetric sonograms. This study was undertaken to determine which factors influence the sonographic visualization of fetal EIFs. METHODS: In part 1 of the study, records from 1,920 fetal sonographic examinations were reviewed for fetal age, indication for sonography, and abnormal findings. For all cases with EIFs recorded and 645 randomly selected cases with no record of EIFs, sonograms were reviewed for heart position at the time of the 4-chamber view, technologist performing the examination, fetal position, heart visibility, transducer frequency, machine type, amount of amniotic fluid, and presence/absence of an EIF. In part 2 of the study, machine settings were evaluated with respect to visualization of EIFs. RESULTS: In part 1 of the study, only the technologist performing the examination and the fetal position were associated with visualization of EIFs. In part 2 of the study, we found that the standard obstetric mode settings are better for visualization of EIFs than are the fetal echocardiographic mode settings. CONCLUSIONS: We conclude that technical factors influence visualization of EIFs. PMID- 11056026 TI - Sonographic demonstration of a pharyngoesophageal diverticulum. AB - We report a case of pharyngoesophageal (Zenker's) diverticulum in a 91-year-old woman. Sonography of the thyroid gland showed diffuse enlargement of the gland and a well-defined, heterogeneous hyperechoic mass that appeared to be in the posterior left lobe. The mass had a smooth hypoechoic wall with a layered appearance anteriorly. Real-time sonography performed during the patient's ingestion of water showed transient changes in the size, margins, and echogenicity of the lesion, which subsequently reverted to its original appearance. PMID- 11056027 TI - Sonographic detection of diffuse peripheral nerve hypertrophy in chronic inflammatory demyelinating polyradiculoneuropathy. AB - Chronic inflammatory demyelinating polyradiculoneuropathy is an autoimmune disease characterized by recurrent demyelination and remyelination with resultant thickening of the peripheral nerves. We report a case in which sonography was instrumental in demonstrating diffuse peripheral nerve hypertrophy. On sonography, both brachial plexuses were found to be diffusely hypertrophic and hypoechoic. Similar findings were noted for the median, sciatic, and femoral nerves. The brachial plexus findings were confirmed by MRI. PMID- 11056028 TI - Sonographic diagnosis of intussusception of the appendix vermiformis. AB - We present 2 cases of intussusception of the appendix vermiformis (IAV) in children. Sonographic examination demonstrated a lead point within a characteristic multiconcentric ring sign, and longitudinal sonograms showed the inverted appendix protruding into the cecal lumen. A contrast-enema study, using a water-soluble contrast medium, was performed in each case, and a "coiled spring" sign or "spiral shell" appearance confirmed the diagnosis. A surgical reduction of the appendix and an appendectomy were performed in each case. PMID- 11056029 TI - Pneumatosis of the bladder wall associated with necrotizing enterocolitis. AB - Increasing use of sonography for imaging the abdomen of neonates has brought greater recognition of its value in diagnosing and monitoring complications of necrotizing enterocolitis. We describe a case in which pneumatosis of the bladder wall was visualized by sonography in a neonate with Down syndrome and necrotizing enterocolitis. PMID- 11056030 TI - Prune-belly syndrome: therapeutic options including in utero placement of a vesicoamniotic shunt. AB - The prune-belly syndrome (PBS) consists of abdominal wall distention with deficiency of the abdominal wall musculature, urinary tract abnormalities, and cryptorchidism. The impaired drainage of the bladder leads to oligohydramnios and pulmonary hypoplasia. We present 4 cases of PBS diagnosed by prenatal sonography. In 2 cases, vesicoamniotic shunt therapy was not indicated because of a poor prognosis based on sonographic and laboratory findings; the pregnancies were terminated. In another case, treatment was not performed because of a twin pregnancy, and the neonate with PBS died the day of delivery by cesarean section at 31 weeks' menstrual age. In the other case, vesicoamniotic shunt therapy was successfully performed, and a healthy child was delivered. Several conditions must be met for vesicoamniotic shunt therapy to have a good chance of success: the karyotype must be normal, other malformations must be excluded by careful sonographic examination, and renal function must be normal, as determined by serial analyses of fetal urine. Generally, the shunt should be inserted as early as possible. PMID- 11056031 TI - Crystal structure and refolding properties of the mutant F99S/M153T/V163A of the green fluorescent protein. AB - The mutant F99S/M153T/V163A of the Green Fluorescent Protein (c3-GFP) has spectral characteristics similar to the wild-type GFP, but it is 42-fold more fluorescent in vivo. Here, we report the crystal structure and the refolding properties of c3-GFP and compare them with those of the less fluorescent wt-GFP and S65T mutant. The topology and the overall structure of c3-GFP is similar to the wild-type GFP. The three mutated residues, Ser99, Thr153, and Ala163, lie on the surface of the protein in three different beta-strands. The side chains of Ser99 and Thr153 are exposed to the solvent, whereas that of Ala163 points toward the interior of the protein. No significant deviation from the structure of the wild-type molecule is found around these positions, and there is not clear evidence of any distortion in the position of the chromophore or of the surrounding residues induced by the mutated amino acids. In vitro refolding experiments on urea-denatured c3-GFP reveal a renaturation behavior similar to that of the S65T molecule, with kinetic constants of the same order of magnitude. We conclude that the higher fluorescence activity of c3-GFP can be attributed neither to particular structural features nor to a faster folding process, as previously proposed. PMID- 11056032 TI - Structural mapping of single cysteine mutants of cardiac troponin I. AB - The global conformation of cardiac muscle troponin I (cTnI) was investigated with single-cysteine mutants by using a combination of sulfhydryl reactivity and fluorescence resonance energy transfer (FRET) to determine cysteine accessibility and intersite distances. The reactivity was determined with a fluorescent reagent for its reaction with cysteine residues singly located at positions 5, 40, 81, 98, 115, 133, 150, 167, and 192. FRET measurements were made by using the endogenous single Trp-192 as the energy donor and an acceptor probe covalently attached to the cysteines as energy acceptor. The results suggest an open and extended conformation of cTnI with a large curvature in which the cysteines are highly exposed to the solvent. These conformational features are largely retained in the segment between residues 40 and 192 upon phosphorylation at Ser-23 and Ser 24. The sulfhydryl groups of the Cys-133 and Cys-150 of the cTnI incorporated into the binary cTnC-cTnI and fully reconstituted troponin complexes experience large reduced exposure resulting from the binding of Ca(2+) to the regulatory site of cTnC, suggesting that key regions of cTnI involved in activation become highly shielded upon activation. In the cTnC-cTnI complex, every intramolecular distance in the cTnI is lengthened and the overall conformation of the bound cTnI remains elongated with reduced exposure for the cysteines. The global conformation of the troponin C-troponin I complex from cardiac muscle has an elongated shape with constrained flexibility. The highly flexible nature of the N terminal extension of cTnI is preserved in the complex, suggesting that this segment of cTnI is either not bound or only loosely bound to the C-domain of cTnC. PMID- 11056033 TI - Receptors coupling to G proteins: is there a signal behind the sequence? AB - Upon the binding of their ligands, G protein-coupled receptors couple to the heterotrimeric G proteins to transduce a signal. One receptor family may couple to a single G protein subtype and another family to several ones. Is there a signal in the receptor sequence that can give an indication of the G protein subtype selectivity? We used a sequence analysis method on biogenic amine and adenosine receptors and concluded that a weak signal can be detected in receptor families where specialization for coupling to a given G protein occurred during a recent divergent evolutionary process. Proteins 2000;41:448-459. PMID- 11056034 TI - Backbone dynamics of barstar: a (15)N NMR relaxation study. AB - Backbone dynamics of uniformly (15)N-labeled barstar have been studied at 32 degrees C, pH 6.7, by using (15)N relaxation data obtained from proton-detected 2D (1)H-(15)N NMR spectroscopy. (15)N spin-lattice relaxation rate constants (R(1)), spin-spin relaxation rate constants (R(2)), and steady-state heteronuclear (1)H-(15)N NOEs have been determined for 69 of the 86 (excluding two prolines and the N-terminal residue) backbone amide (15)N at a magnetic field strength of 14.1 Tesla. The primary relaxation data have been analyzed by using the model-free formalism of molecular dynamics, using both isotropic and axially symmetric diffusion of the molecule, to determine the overall rotational correlation time (tau(m)), the generalized order parameter (S(2)), the effective correlation time for internal motions (tau(e)), and NH exchange broadening contributions (R(ex)) for each residue. As per the axially symmetric diffusion, the ratio of diffusion rates about the unique and perpendicular axes (D( parallel)/D( perpendicular)) is 0.82 +/- 0.03. The two results have only marginal differences. The relaxation data have also been used to map reduced spectral densities for the NH vectors of these residues at three frequencies: 0, omega(H), and omega(N), where omega(H),(N) are proton and nitrogen Larmor frequencies. The value of tau(m) obtained from model-free analysis of the relaxation data is 5.2 ns. The reduced spectral density analysis, however, yields a value of 5.7 ns. The tau(m) determined here is different from that calculated previously from time resolved fluorescence data (4.1 ns). The order parameter ranges from 0.68 to 0.98, with an average value of 0.85 +/- 0.02. A comparison of the order parameters with the X-ray B-factors for the backbone nitrogens of wild-type barstar does not show any considerable correlation. Model-free analysis of the relaxation data for seven residues required the inclusion of an exchange broadening term, the magnitude of which ranges from 2 to 9.1 s(-1), indicating the presence of conformational averaging motions only for a small subset of residues. PMID- 11056035 TI - Covariance analysis of protein families: the case of the variable domains of antibodies. AB - A nonrestrictive method for identifying covariance in protein families is described and applied to human and mouse germline Vkappa and VH sequence alignments. Amino acids that occur at each position in a sequence alignment are divided into two sets, called a word, by generating all possible combinations of alternative amino acids. Each word is associated with a pattern of changes. Words with identical patterns identify covariant positions. In antibody variable domains, the number of words generated ranged between 1103 and 2195 depending on the alignment, of which 4 to 12 % occurred in covariant pairs. Despite the nonrestrictive character of pattern generation, covariant residues did not reflect a random selection with respect to the nature of amino acid changes and/or their spatial proximity in a reference crystallographic structure. This approach allowed the identification of a covariance signal for positions with high variability, mostly located in the outer part of the common structural framework of antibody variable domains. Covariance in these regions may reflect the existence of alternative and mutually exclusive atomic arrangements that are compatible with antibody function. The method may be of general applicability to rationalize residue variability in protein families. PMID- 11056036 TI - Fluctuations between stabilizing and destabilizing electrostatic contributions of ion pairs in conformers of the c-Myc-Max leucine zipper. AB - In solution proteins often exhibit backbone and side-chain flexibility. Yet electrostatic interactions in proteins are sensitive to motions. Hence, here we study the contribution of ion pairs toward protein stability in a range of conformers which sample the conformational space in solution. Specifically, we focus on the electrostatic contributions of ion pairs to the stability of each of the conformers in the NMR ensemble of the c-Myc-Max leucine zipper and to their average energy minimized structure. We compute the electrostatic contributions of inter- and intra-helical ion pairs and of an ion pair network. We find that the electrostatic contributions vary considerably among the 40 NMR conformers. Each ion pair, and the network, fluctuates between being stabilizing and being destabilizing. This fluctation reflects the variability in the location of the ion pairing residues and in the geometric orientation of these residues, both with respect to each other and with respect to other charged groups in the rest of the protein. Ion pair interactions in the c-Myc-Max leucine zipper in solution depend on the protein conformer which is analyzed. Hence, the overall stabilizing (or destabilizing) contribution of an ion pair is conformer population-dependent. This study indicates that free energy calculations performed using the continuum electrostatics methodology are sensitive to protein conformational details. PMID- 11056037 TI - Optimization of a new score function for the detection of remote homologs. AB - The growth in protein sequence data has placed a premium on ways to infer structure and function of the newly sequenced proteins. One of the most effective ways is to identify a homologous relationship with a protein about which more is known. While close evolutionary relationships can be confidently determined with standard methods, the difficulty increases as the relationships become more distant. All of these methods rely on some score function to measure sequence similarity. The choice of score function is especially critical for these distant relationships. We describe a new method of determining a score function, optimizing the ability to discriminate between homologs and non-homologs. We find that this new score function performs better than standard score functions for the identification of distant homologies. PMID- 11056038 TI - A novel measure characterized by a polar energy surface approximation for recognition and classification of transmembrane protein structures. AB - A new theoretical method has been developed for recognition and classification of membrane proteins. The method is based on computation of a polar energy surface that can reveal characteristic interaction patterns for individual helices even if crystal or NMR structure coordinates are not available. A protein with N transmembrane helices is described as a set of N vectors that are derived from a Fourier analysis of this polar energy surface computed for each helix. We then derive a polarity difference score (PDS) for any two proteins computed as the root mean square deviation between the respective vector coordinate sets. The score was found to correlate with the degree of structural similarity between the following three protein families for which tertiary structures have been determined: bacteriorhodopsin, rhodopsin, and the cytochrome c oxidase III subunit. PMID- 11056039 TI - Discrimination of near-native protein structures from misfolded models by empirical free energy functions. AB - Free energy potentials, combining molecular mechanics with empirical solvation and entropic terms, are used to discriminate native and near-native protein conformations from slightly misfolded decoys. Since the functional forms of these potentials vary within the field, it is of interest to determine the contributions of individual free energy terms and their combinations to the discriminative power of the potential. This is achieved in terms of quantitative measures of discrimination that include the correlation coefficient between RMSD and free energy, and a new measure labeled the minimum discriminatory slope (MDS). In terms of these criteria, the internal energy is shown to be a good discriminator on its own, which implies that even well-constructed decoys are substantially more strained than the native protein structure. The discrimination improves if, in addition to the internal energy, the free energy expression includes the electrostatic energy, calculated by assuming non-ionized side chains, and an empirical solvation term, with the classical atomic solvation parameter model providing slightly better discrimination than a structure-based atomic contact potential. Finally, the inclusion of a term representing the side chain entropy change, and calculated by an established empirical scale, is so inaccurate that it makes the discrimination worse. It is shown that both the correlation coefficient and the MDS value (or its dimensionless form) are needed for an objective assessment of a potential, and that together they provide much more information on the origins of discrimination than simple inspection of the RMSD-free energy plots. PMID- 11056040 TI - Predictions of protein segments with the same aminoacid sequence and different secondary structure: a benchmark for predictive methods. AB - The most stringent test for predictive methods of protein secondary structure is whether identical short sequences that are known to be present with different conformations in different proteins known at atomic resolution can be correctly discriminated. In this study, we show that the prediction efficiency of this type of segments in unrelated proteins reaches an average accuracy per residue ranging from about 72 to 75% (depending on the alignment method used to generate the input sequence profile) only when methods of the third generation are used. A comparison of different methods based on segment statistics (2nd generation methods) and/or including also evolutionary information (3rd generation methods) indicate that the discrimination of the different conformations of identical segments is dependent on the method used for the prediction. Accuracy is similar when methods similarly performing on the secondary structure prediction are tested. When evolutionary information is taken into account as compared to single sequence input, the number of correctly discriminated pairs is increased twofold. The results also highlight the predictive capability of neural networks for identical segments whose conformation differs in different proteins. PMID- 11056041 TI - Nucleoside binding site of herpes simplex type 1 thymidine kinase analyzed by X ray crystallography. AB - The crystal structures of the full-length Herpes simplex virus type 1 thymidine kinase in its unligated form and in a complex with an adenine analogue have been determined at 1.9 A resolution. The unligated enzyme contains four water molecules in the thymidine pocket and reveals a small induced fit on substrate binding. The structure of the ligated enzyme shows for the first time a bound adenine analogue after numerous complexes with thymine and guanine analogues have been reported. The adenine analogue constitutes a new lead compound for enzyme prodrug gene therapy. In addition, the structure of mutant Q125N modifying the binding site of the natural substrate thymidine in complex with this substrate has been established at 2.5 A resolution. It reveals that neither the binding mode of thymidine nor the polypeptide backbone conformation is altered, except that the two major hydrogen bonds to thymidine are replaced by a single water mediated hydrogen bond, which improves the relative acceptance of the prodrugs aciclovir and ganciclovir compared with the natural substrate. Accordingly, the mutant structure represents a first step toward improving the virus-directed enzyme-prodrug gene therapy by enzyme engineering. PMID- 11056042 TI - pK(a) calculations of calbindin D(9k): effects of Ca(2+) binding, protein dielectric constant, and ionic strength. AB - Calbindin is a small (75 residues) helix-loop-helix ("EF-hand") calcium-binding protein belonging to the calmodulin superfamily. It binds two Ca(2+) ions. Continuum electrostatics in combination with the boundary element method was employed for the calculation of the acid-dissociation constants K(a) (pK(a) = log K(a)) values of all titratable residues in the protein. The objectives were to determine quantitatively the effects of divalent ion binding and small ion induced structural changes on predicted pK(a)'s. Computations were carried out for the apo and holo form of calbindin, for which both X-ray and NMR structures were available. Comparison was made with several sets of experimental pK(a) values determined by NMR spectroscopy. Different choices of the dielectric constant (ranging from 4 to 78.5) for calbindin and variations in ionic strength (from 0 to 0.3 M) were investigated in a systematic fashion. Removal of the two bound Ca(2+) ions increases the pK(a) values of all residues if no conformational changes were allowed. If conformational differences between the apo and holo were accounted for, shifts in either direction were observed. Titrating groups that are directly involved in Ca(2+) binding (Asp and Glu) required a dielectric constant of 78.5 for the holo structure to obtain a reasonable estimate of their pK(a)'s. For the apo structure, passable values for the pK(a)'s of these ligating groups could be determined if the structure was allowed to relax upon ion removal. PMID- 11056043 TI - Mifepristone (RU 486). PMID- 11056044 TI - Colesevelam (Welchol) for hypercholesterolemia. PMID- 11056045 TI - Identification of differentially expressed genes in normal and tumor human gastric tissue. AB - The search for differentially expressed genes in gastric cancer may help define molecular alterations and molecular diagnosis of gastric cancer. Using the differential display PCR technique, we identified 18 genes that are differentially expressed between normal and tumor human gastric tissues. Their expressions were verified with reverse Northern blot analysis and Northern blot analysis. Oxidative phosphorylation-related genes, antizyme inhibitor of ornithine decarboxylase, protein phosphatase-1beta, 35-kDa peroxisomal membrane protein, and cystic fibrosis transmembrane conductance receptor were highly expressed in tumor tissue, whereas pepsinogen A, Na-K ATPase alpha subunit, nerve growth factor receptor, and alpha-tropomyosin were highly expressed in normal tissue. In addition, 3 unknown genes were found to be differentially expressed in paired gastric tissues. These differentially expressed genes may provide significant opportunities for further understanding of gastric carcinogenesis and the molecular diagnosis of gastric cancer. PMID- 11056046 TI - High-throughput scanning of the rat genome using interspersed repetitive sequence PCR markers. AB - We report the establishment of a hybridization-based marker system for the rat genome based on the PCR amplification of interspersed repetitive sequences (IRS). Overall, 351 IRS markers were mapped within the rat genome. The IRS marker panel consists of 210 nonpolymorphic and 141 polymorphic markers that were screened for presence/absence polymorphism patterns in 38 different rat strains and substrains that are commonly used in biomedical research. The IRS marker panel was demonstrated to be useful for rapid genome screening in experimental rat crosses and high-throughput characterization of large-insert genomic library clones. Information on corresponding YAC clones is made available for this IRS marker set distributed over the whole rat genome. The two existing rat radiation hybrid maps were integrated by placing the IRS markers in both maps. The genetic and physical mapping data presented provide substantial information for ongoing positional cloning projects in the rat. PMID- 11056047 TI - Tissue-specific expression of antisense and sense transcripts at the imprinted Gnas locus. AB - The mouse Gnas gene encodes an important signal transduction protein, the alpha subunit of the stimulatory G protein, G(s). In humans, partial deficiency of G(s)alpha, the alpha subunit of G(s), results in the hormone-resistance syndrome pseudohypoparathyroidism type 1a. The mouse Gnas (and the human GNAS1) locus is transcribed from three promoter regions. Transcripts from P1, which encode Nesp55, are derived from the maternal allele only. Transcripts from P2 encode Xlalphas and are derived only from the paternal allele, while transcripts from P3 encode the alpha subunit and are from both parental alleles. The close proximity of reciprocal imprinting suggests the presence of important putative imprinting elements in this region. In this report, we demonstrate that the reciprocal imprinting occurs in normal tissues of interspecific (Mus spretus x C57BL/6) mice. Transcripts from P1 are most abundant in CNS (pons and medulla) in contrast to the more ubiquitous expression from P2 and P3. In the P1-P2 genomic region, we have identified an antisense transcript that starts 2.2 kb upstream of the P2 exon and spans the P1 region. While the P1 transcript is derived from the maternal allele, the P1-antisense (Gnas-as) is derived only from the paternal allele in most but not all tissues. Although both the Nesp55 region and the Gnas as transcripts are present in cerebral cortex, adrenal, and spleen, Gnas-as is abundant in some tissues in which transcription from the Nesp55 region is negligible. Furthermore, the Nesp55 region transcripts remain strictly imprinted in tissues that lack Gnas-as. Our results suggest that multiple imprinting elements, including the unique Gnas-as, regulate the allelic expression of the Nesp55 region sense transcript. PMID- 11056048 TI - A sequence-ready PAC contig of a 550-kb region on rat chromosome 4 including the diabetes susceptibility gene Lyp. AB - The Lyp locus controls diabetes development in rats. The diabetogenic allele in diabetes-prone BB rats is responsible for T cell lymphopenia characterized by the absence of regulatory T cells. We present refined genetic and radiation hybrid maps of the Lyp region on rat chromosome 4, a single 800-kb rat yeast artificial chromosome and a rat P1-derived artificial chromosome (PAC) contig corresponding to approximately 550 kb, both encompassing the entire candidate region. The contig, consisting of 48 PACs, gives 3- to 12-fold coverage. Genetic, radiation hybrid, and physical data were all in agreement and supported the same marker order. Nine genes and ESTs were identified in the contig in addition to a rat EST from the University of Iowa rat EST database-all possible candidate genes for Lyp. Alignment of our rat PAC contig with sequenced human PAC/BAC contigs confirms the position within the region of 3 of the 10 candidates and identifies an additional 8 genes/ESTs as candidates. These data will facilitate identification of Lyp. PMID- 11056049 TI - A novel G-protein-coupled receptor gene expressed in striatum. AB - Differential display screening for region-specific transcripts in rat brain revealed a novel striatum-specific transcript encoding an orphan G-protein coupled receptor (GPCR) designated Strg/Gpr88 for striatum-specific GPCR. We isolated its homologues from human (HGMW-approved symbol GPR88) and mouse and mapped them to chromosomes 1p21.3 and 3G1, respectively. These loci are syntenic to each other, thereby suggesting their orthology. The predicted primary sequences of Strg/Gpr88 proteins are highly conserved between human and rodents and show the highest level of homology to receptors for biogenic amines. However, Strg/Gpr88 lacks some residues conserved in all known biogenic amine receptors and hence may represent a novel subtype of GPCR. Northern blot and in situ hybridization analyses revealed that Strg/Gpr88 transcripts are expressed almost exclusively in striatum in both human and rodents. Remarkable conservation in primary structure and a unique expression pattern may indicate a role for Strg/Gpr88 in the fundamental functions of striatum such as the control of motor behavior. PMID- 11056050 TI - Novel human esophagus-specific gene c1orf10: cDNA cloning, gene structure, and frequent loss of expression in esophageal cancer. AB - We have identified a novel human gene, designated C1orf10, using modified differential display PCR. The C1orf10 gene, which spans 5 kb in length, is composed of three exons. The deduced protein contains 495 amino acids with one transmembrane domain. The amino acid sequence of C1orf10 is characterized by the presence of a calcium-binding motif of about 90 amino acids at its N-terminal and a conserved consecutive repeat sequence of 60 amino acids that was identified previously only in bacterial ice nucleation proteins. In normal adult tissues, C1orf10 is highly expressed only in the esophagus and was undetectable in a total of 15 other tissues examined, suggesting its important role in esophageal cells. The expression of C1orf10 is either dramatically reduced or absent in esophageal cancer cell lines (3/3) as well as primary esophageal cancer tissues (35/37) compared with the corresponding normal esophageal mucosa. Using a radiation hybrid panel, C1orf10 was found to be located on chromosome 1q21. These findings suggest that expression of C1orf10 is unique to esophageal cells and that loss of its expression may play a role in the development of esophageal cancer. PMID- 11056051 TI - KLK12 is a novel serine protease and a new member of the human kallikrein gene family-differential expression in breast cancer. AB - Kallikreins are a subgroup of serine proteases that are involved in the posttranslational processing of polypeptide precursors. Growing evidence suggests that many kallikreins are implicated in carcinogenesis. In rodents, kallikreins are encoded by a large multigene family, but in humans, only three genes have been identified. By using the positional candidate approach, we were able to identify a new kallikrein-like gene, tentatively named KLK12 (for kallikrein gene 12). This new gene maps to chromosome 19q13.3-q13.4, is formed of five coding exons, and shows structural similarity to serine proteases and other known kallikreins. KLK12 is expressed in a variety of tissues including salivary gland, stomach, uterus, lung, thymus, prostate, colon, brain, breast, thyroid, and trachea. We identified three splicing forms of KLK12 that are expressed in many tissues. Our preliminary results indicate that the expression of KLK12 is down regulated at the mRNA level in breast cancer tissues and is up-regulated by steroid hormones in breast and prostate cancer cell lines. This gene may be involved in the pathogenesis and/or progression of certain cancer types and may find applicability as a novel cancer biomarker. PMID- 11056052 TI - Cloning of TC-1 (C8orf4), a novel gene found to be overexpressed in thyroid cancer. AB - A novel gene highly expressed in thyroid cancer, designated TC-1 (thyroid cancer 1), was cloned from suppression subtractive hybridization between papillary thyroid carcinoma and its surrounding normal thyroid tissue. Overexpression of TC 1 in thyroid cancer was confirmed in 15/16 paired samples by RT-PCR and Northern analysis. Ubiquitously expressed in human tissues, the TC-1 sequence showed no homology to any known gene, but matched a cluster of ESTs. After alignment of our sequence with the ESTs, the missing transcription start site was obtained by 5' RACE and verified by primer extension analysis. The full-length mRNA sequence of 1327 bp has an open reading frame of 321 bp, which encodes a highly conserved protein. Three regulatory motifs were identified at the expected positions within 1 kb of the 5' flanking sequence obtained by genome walking. Using fluorescence in situ hybridization, TC-1 was localized to chromosome 8p11.2. The overexpression of TC-1 in papillary carcinoma suggests that it may have an important role in thyroid carcinogenesis. PMID- 11056053 TI - Mapping, characterization, and expression analysis of the SM-20 human homologue, c1orf12, and identification of a novel related gene, SCAND2. AB - Major psychosis was shown to segregate with a balanced translocation (1q42.1; 11q14.3) in a multigenerational family. This study describes the identification of a human SM-20 homologue gene that lies at about 400 kb on the centromeric side of the 1q42.1 breakpoint. The full-length cDNA sequence and gene structure were determined. Expression analysis was performed, showing high expression levels in skeletal and cardiac muscles; in the central nervous system, expression was restricted to dopaminergic neurons and spinal motoneurons. A second gene displaying high sequence similarity with SM-20 was also identified by BLAST. This gene, located on chromosome 15, is likely to have evolved by retroposition of SM 20 mRNA and an exon-shuffling mechanism. It encodes a 306-amino-acid protein harboring strong homology with an N-terminal motif found in some zinc-finger proteins. This gene was named SCAND2 (SCAN domain-containing 2). PMID- 11056054 TI - Cloning and characterization of two mouse genes with homology to the yeast Sir2 gene. AB - The yeast Sir2 gene encodes a protein (Sir2p) that plays an essential role in silencing regulation at mating-type loci, rDNA, and telomeres. Recent studies have also shown that the protein participates in cell cycle regulation, DNA double-strand break repair, meiotic checkpoint control, and histone deacetylation. Overexpression of wildtype Sir2p in yeast resulted in an extended life span but mutant Sir2p shortened the life span, suggesting its function in aging processes. Sir2p is evolutionarily conserved from prokaryotes to higher eukaryotes. However, its function(s) in mammals remains unknown. To investigate Sir2p function(s) in mice, we cloned and characterized two mouse Sir2-like genes. Our results revealed that the two mouse Sir2-like proteins (mSIR2L2 and mSIR2L3) are most similar to the human Sir2-like proteins SIR2L2 and SIR2L3, respectively. Sir2 core domains are highly conserved in the two proteins and yeast Sir2p; however, the intracellular localizations of both mSIR2L2 and mSIR2L3 differ from that of yeast Sir2p and from one another. The two mouse genes have completely different genomic structures but were mapped on the same chromosome. It seems that the two mouse proteins, though they have Sir2 conserved domains, may function differently than yeast Sir2p. PMID- 11056055 TI - Genomic structure of the mouse Ap3b1 gene in normal and pearl mice. AB - The mouse hypopigmentation mutant pearl is an established model for Hermansky Pudlak syndrome (HPS), a genetically heterogenous disease with misregulation of the biogenesis/function of melanosomes, lysosomes, and platelet dense granules. The pearl (Ap3b1) gene encodes the beta3A subunit of the AP-3 adaptor complex, which regulates vesicular trafficking. The genomic structure of the normal Ap3b1 gene includes 25 introns and a putative promoter sequence. The original pearl (pe) mutation, which has an unusually high reversion rate on certain strain backgrounds, has been postulated to be caused by insertion of a transposable element. Indeed, the mutation contains a 215-bp partial mouse transposon at the junction point of a large tandem genomic duplication of 6 exons and associated introns. At the cDNA level, three pearl mutations (pearl, pearl-8J, and pearl-9J) are caused by deletions or duplications of a complete exon(s). PMID- 11056056 TI - Genomic organization of the JEM-1 (BLZF1) gene on human chromosome 1q24: molecular cloning and analysis of its promoter region. AB - The Jem-1 (JEM-1, HGMW-approved symbol BLZF1) gene mapping to human chromosome 1q24 codes for a ubiquitously expressed 3-kb mRNA, translated in a 45-kDa nuclear protein. Recent studies have shown a deficient expression of this gene in acute promyelocytic leukemia (APL). However, treatment with retinoids was able to upregulate JEM-1 mRNA in maturing NB4 leukemia cells. Here, we report the characterization of the structural organization of JEM-1. By hybridization screening of a human genomic library derived from blood mononuclear cells, five overlapping genomic DNA clones were isolated. These clones extend over 34 kb of the human genome and comprise the complete JEM-1 gene and a 4-kb 5'flanking region. Determination of the exon-intron structure of Jem-1 revealed seven exons whose junctions with introns exhibited typical splice sequences. A shorter transcript (Jem-1s, 1.3 kb) generated by exon 3 extension and polyadenylation was identified. Its translation generated a 23-kDa protein that exhibited a cytoplasmic localization. 5'RACE-PCR identified a major transcription start site (TSS) located at 403 nt upstream of the ATG. Computer analysis of the 1. 8-kb 5'flanking region showed that it lacks a TATA box, Inr motifs or DPE motifs, but it contains a typical CCAAT box located 95 bp upstream of the TSS. Sequencing also revealed potential cis-acting elements for multiple transcription regulators including Sp1, GATA, C/EBP, AP-1, and Pu1. No retinoic acid receptor elements or retinoic X receptor elements were detected. This 1.8-kb DNA sequence showed a strong constitutive promoter activity determined by a luciferase-reporter gene assay in transiently transfected HeLa cells. Retinoids further increased luciferase expression 2.7-fold. We demonstrated that the 1-kb distal sequence contains yet unidentified elements reducing constitutive transcription. Thus, the maximal constitutive promoter activity was assigned to a -432 + 101 region overlapping the TSS. These data support the idea of a constitutive expression of JEM-1, but a negative regulation in APL released by retinoids. PMID- 11056057 TI - A frequent deletion polymorphism on chromosome 22q13 identified by representational difference analysis of ovarian cancer. AB - Sizable homozygous deletions (>100 bp) of genomic DNA in cancer cells are typically interpreted as an indication of the location of a tumor suppressor gene. In an effort to identify novel ovarian growth-suppressing genes, we performed representational difference analysis (RDA) of ovarian cancer cells. One of the RDA probes identified a 276-bp region of chromosome 22q deleted in 47% of the ovarian cancer cell lines examined. This small deletion was also found in the genomic DNA of 25% of colon cancer cell lines examined and, surprisingly, in 18% of the blood DNA samples from healthy controls. The deleted allele, which was named u22q, has a frequency of approximately 50% in the population and is in Hardy-Weinberg equilibrium with the intact allele. The deleted DNA sequence is flanked by direct repeats and likely originated through a slipped mispairing mechanism. The deletion did not encompass known transcripts or expressed sequence tags. It therefore appears likely that u22q represents a common polymorphism, often hemizygous in ovarian cancer because of a high rate of LOH of chromosome 22q. These findings provide an example of a sizable homozygous deletion that does not appear to be associated with disease. Such a finding provides a cautionary tale for positional cloning projects initiated exclusively on the basis of the identification of homozygous deletions. The possibility that the deletions in question may be constitutive should always be considered since it is probable that the genome contains a large number deletions/insertions of various sizes. PMID- 11056058 TI - Bioinformatics and metabolic engineering PMID- 11056059 TI - The emergence of pattern discovery techniques in computational biology. AB - In the past few years, pattern discovery has been emerging as a generic tool of choice for tackling problems from the computational biology domain. In this presentation, and after defining the problem in its generality, we review some of the algorithms that have appeared in the literature and describe several applications of pattern discovery to problems from computational biology. PMID- 11056060 TI - A comparative study of global stress gene regulation in response to overexpression of recombinant proteins in Escherichia coli. AB - Global gene regulation throughout the Escherichia coli stress response to overexpression of each of five recombinant proteins was evaluated. Reverse transcriptase polymerase chain reaction-amplified mRNA from induced and control cells were hybridized with a DNA array of Kohara clones representing 16% (700 genes) of the E. coli genome. Subsequently, Northern analysis was performed for quantification of specific gene dynamics and statistically significant overlap in the regulation of 11 stress-related genes was found using correlation analysis. The results reported here establish that there are dramatic changes in the transcription rates of a broad range of stress genes (representing multiple regulons) after induction of recombinant protein. Specifically, the responses included significantly increased upregulation of heat shock (ftsH, clpP, lon, ompT, degP, groEL, aceA, ibpA), SOS/DNA damage (recA, lon, IS5 transposase), stationary phase (rpoS, aceA), and bacteriophage life cycle (ftsH, recA) genes. Importantly, similarities at the microscopic (gene) level were not clearly reflected at the macroscopic (growth rate, lysis) level. The use of such dynamic data is critical to the design of gene-based sensors, the engineering of metabolic pathways, and the determination of parameters (harvest and induction times) needed for successful recombinant E. coli fermentations. PMID- 11056061 TI - The organization of metabolic reaction networks: a signal-oriented approach to cellular models. AB - Complex metabolic networks are characterized by a great number of elements and many regulatory loops. The description of these networks with mathematical models requires the definition of functional units that group together several cellular processes. The approach presented here is based on the idea that cellular functional units may be assigned directly to mathematical modeling objects. Because the proposed modeling objects have defined inputs and outputs, they can be connected with other modeling objects until eventually the whole metabolism is covered. This modular approach guarantees a high transparency for biologists as well as for engineers. Three criteria are introduced to demarcate functional units. The criteria consider the physiological pathways, the organization of the corresponding genes, and the observation that cellular systems can be structured into units showing a hierarchy of signal transduction and processing. As an example, the carbon catabolic reactions in Escherichia coli are discussed as members of a functional unit catabolism. PMID- 11056062 TI - DNA microarray detection of metabolic responses to protein overproduction in Escherichia coli. AB - It has been commonly observed that gratuitous overexpression of proteins in Escherichia coli causes growth retardation. However, the molecular events involved in the metabolic response to the over-expression of proteins are still unclear. Here we used DNA microarray technology to characterize the changes in transcriptional patterns of selected host genes during protein overexpression. A nontoxic, soluble protein, LuxA (coded by luxA), which is the alpha-subunit of the luciferase heterodimer, was overexpressed for this purpose. A total of 132 E. coli genes, including those in the central metabolism, key biosynthetic pathways, and selected regulatory functions, were used as probes for detecting the level of mRNA transcripts in E. coli strains JM109, MC4100, and VJS676A during protein overexpression. Upon induction, these strains shared several common responses, such as the upregulation of glk and the heat shock genes as well as the downregulation of fba, ppc, atpA, and gnd. In addition, the biosynthesis genes glnA, glyA, and leuA were downregulated in all three strains. Media-dependent responses were also observed in our study. For example, many respiratory genes that were upregulated in defined media showed an opposite effect in complex media under protein-overproducing conditions. These results demonstrate that gratuitous overexpression of proteins triggers a complex global response that involves several metabolic and regulatory systems. Explanations based on either existing knowledge of global regulations such as the heat shock response and the stringent response or stoichiometric analysis without regulatory considerations cannot account for the response induced by protein overexpression. PMID- 11056063 TI - Patterns of regulation from mRNA and protein time series. AB - The rapid advance of genome sequencing projects challenges biologists to assign physiological roles to thousands of unknown gene products. We suggest here that regulatory functions and protein-protein interactions involving specific products may be inferred from the trajectories over time of their mRNA and free protein levels within the cell. The level of a protein in the cytoplasm is governed not only by the level of its mRNA and the rate of translation, but also by the protein's folding efficiency, its biochemical modification, its complexation with other components, its degradation, and its transport from the cytoplasmic space. All these co- and post translational events cause the concentration of the protein to deviate from the level that would result if we only accounted for translation of its mRNA. The dynamics of such deviations can create patterns that reflect regulatory functions. Moreover, correlations among deviations highlight protein pairs involved in potential protein-protein interactions. We explore and illustrate these ideas here using a genetically structured simulation for the intracellular growth of bacteriophage T7. PMID- 11056064 TI - Mining of biological data I: identifying discriminating features via mean hypothesis testing. AB - Large volumes of data are routinely collected during bioprocess operations and, more recently, in basic biological research using genomics-based technologies. While these data often lack sufficient detail to be used for mechanism identification, it is possible that the underlying mechanisms affecting cell phenotype or process outcome are reflected as specific patterns in the overall or temporal sensor logs. This raises the possibility of identifying outcome-specific fingerprints that can be used for process or phenotype classification and the identification of discriminating characteristics, such as specific genes or process variables. The aim of this work is to provide a systematic approach to identifying and modeling patterns in historical records and using this information for process classification. This approach differs from others in that emphasis is placed on analyzing the data structure first and thereby extracting potentially relevant features prior to model creation. The initial step in this overall approach is to first identify the discriminating features of the relevant measurements and time windows, which can then be subsequently used to discriminate among different classes of process behavior. This is achieved via a mean hypothesis testing algorithm. Next, the homogeneity of the multivariate data in each class is explored via a novel cluster analysis technique called PC1 Time Series Clustering to ensure that the data subsets used accurately reflect the variability displayed in the historical records. This will be the topic of the second paper in this series. We present here the method for identifying discriminating features in data via mean hypothesis testing along with results from the analysis of case studies from industrial fermentations PMID- 11056065 TI - Mining of biological data II: assessing data structure and class homogeneity by cluster analysis. AB - An important step in data analysis is class assignment which is usually done on the basis of a macroscopic phenotypic or bioprocess characteristic, such as high vs low growth, healthy vs diseased state, or high vs. low productivity. Unfortunately, such an assignment may lump together samples, which when derived from a more detailed phenotypic or bioprocess description are dissimilar, giving rise to models of lower quality and predictive power. In this paper we present a clustering algorithm for data preprocessing which involves the identification of fundamentally similar lots on the basis of the extent of similarity among the system variables. The algorithm combines aspects of cluster analysis and principal component analysis by applying agglomerative clustering methods to the first principal component of the system data matrix. As part of a rational strategy for developing empirical models, this technique selects lots (samples) which are most appropriate for inclusion in a training set by analyzing multivariate data homogeneity. Samples with similar data structures are identified and grouped together into distinct clusters. This knowledge is used in the formation of potential training sets. Additionally, this technique can identify atypical lots, i.e., samples that are not simply outliers but exhibit the general properties of one class but have been given the assignment of the other. The method is presented along with examples from its application to fermentation data sets. PMID- 11056066 TI - Dynamics of gene expression in rat hepatocytes under stress. AB - The response of cells to physical or biochemical stress involves concerted changes in the expression of a large number of genes encoding various functions. We have used a quantitative kinetic RT-PCR technique to follow the dynamics of changes in transcription factor and acute-phase mRNA levels in cultured rat hepatocytes subjected to either elevated temperature (40 degrees C) or exposure to the inflammatory cytokine interleukin-6. The profiles of transcription factor gene expression displayed rapid and coordinate regulation, attainment of new steady-states, transitions in some instances from up-regulation to down regulation (or vice versa), and, for elevated temperature, multiple spikes of up regulation. Transcripts of acute-phase genes generally displayed relatively small changes during the first few hours followed by more significant changes over the course of tens of hours (elevated temperature) to days (IL-6 exposure). These observations are all consistent with the notion of genetic reprogramming due to a network of interacting transcription factor proteins and transcripts. We utilized a simple transcription/translation model incorporating autoregulation to describe the dynamics of transcription factor gene expression. This model successfully described key features of the transcription factor dynamics, most notably the multiple spikes observed after exposure to elevated temperature. The dynamics of gene expression are rich in information that, with considerably more study, may eventually be exploited to provide insights into the interplay of genetic networks in regulating a variety of cellular responses. PMID- 11056067 TI - Tendency modeling: a new approach to obtain simplified kinetic models of metabolism applied to Saccharomyces cerevisiae. AB - A novel approach to construct kinetic models of metabolic pathways, to be used in metabolic engineering, is presented: the tendency modeling approach. This approach greatly facilitates the construction of these models and can easily be applied to complex metabolic networks. The resulting models contain a minimal number of parameters; identification of their values is straightforward. Use of in vitro obtained information in the identification of the kinetic equations is minimized. The tendency modeling approach has been used to derive a dynamic model of primary metabolism for aerobic growth of Saccharomyces cerevisiae on glucose, in which compartmentation is included. Simulation results obtained with the derived model are satisfying for most of the carbon metabolites that have been measured. Compared to a more detailed model, the simulations of our model are less accurate, but taking into account the much smaller number of kinetic parameters (35 instead of 84), the tendency the modeling approach is considered promising. PMID- 11056068 TI - Cognitive Load and the Equality Heuristic: A Two-Stage Model of Resource Overconsumption in Small Groups. AB - Two studies were conducted to test a two-stage model of the psychological mechanisms underlying the overconsumption of scarce resources in small groups. The model proposes that members of groups sharing resources first anchor their consumption choices on an "equal-division" heuristic and then, given sufficient cognitive capacity, adjust their choices in a self-serving direction. The results from both studies support the model. The first study found that when faced with a common resource pool almost all group members expressed thoughts regarding equality; however, individuals with sufficient cognitive capacity requested more from the pool and expressed more task-relevant thoughts than individuals lacking the necessary cognitive resources. The second study provided additional support for the two-stage model by demonstrating that group members' cognitions occur in the order predicted by the model and by demonstrating that an individual difference, social value orientation, affects thought processes only when individuals are not experiencing high cognitive loads. Implications are discussed. Copyright 2000 Academic Press. PMID- 11056069 TI - Who Takes the Lead in Risky Decision Making? Effects of Group Members' Risk Preferences and Prototypicality. AB - In two experiments, we studied the effects of (a) the extent to which group members are risk seeking in comparison with others in the group and (b) group member prototypicality (the extent to which individuals hold group-typical risk preferences) on the likelihood that group members will take the lead in risky decision making. Participants were led to believe that they engaged in a four person group discussion and received bogus feedback about their own risk preferences, the risk preferences of the other group members, and the risk preferences of their group as a whole. In Experiment 2, we also manipulated the framing of the decision problem (gain vs. loss frame). Results supported the hypotheses that (a) more risk seeking members are more likely to take the lead and (b) prototypical members are more likely to take the lead when the problem facing the group is ambiguous (i.e., when group risk preferences and decision framing are incongruent), whereas nonprototypical members are more likely to take the lead when the problem facing the group is relatively clear-cut (i.e., when group risk preferences and decision framing are congruent). Copyright 2000 Academic Press. PMID- 11056070 TI - Deception and Retribution in Repeated Ultimatum Bargaining. AB - This paper investigates the dynamics of deception and retribution in repeated ultimatum bargaining. Anonymous dyads exchanged messages and offers in a series of four ultimatum bargaining games that had prospects for relatively large monetary outcomes. Variations in each party's knowledge of the other's resources and alternatives created opportunities for deception. Revelation of prior unknowns exposed deceptions and created opportunities for retribution in subsequent interactions. Results showed that although proposers and responders chose deceptive strategies almost equally, proposers told more outright lies. Both were more deceptive when their private information was never revealed, and proposers were most deceptive when their potential profits were largest. Revelation of proposers' lies had little effect on their subsequent behavior even though responders rejected their offers more than similar offers from truthful proposers or proposers whose prior deceit was never revealed. The discussion and conclusions address the dynamics of deception and retribution in repeated bargaining interactions. Copyright 2000 Academic Press. PMID- 11056071 TI - Advice Taking in Decision Making: Egocentric Discounting and Reputation Formation. AB - Our framework for understanding advice-taking in decision making rests on two theoretical concepts that motivate the studies and serve to explain the findings. The first is egocentric discounting of others' opinions and the second is reputation formation for advisors. Advice discounting is attributed to differential information, namely, the notion that decision makers have privileged access to their internal reasons for holding their own opinion, but not to the advisors' internal reasons. Reputation formation is related to the negativity effect in impression formation and to the trust asymmetry principle. In three studies we measured decision makers' weighting policy for advice and, in a fourth study, their willingness to pay for it. Briefly, we found that advice is discounted relative to one's own opinion, while advisors' reputations are rapidly formed and asymmetrically revised. The asymmetry implies that it may be easier for advisors to lose a good reputation than to gain one. The cognitive and social origins of these phenomena are considered. Copyright 2000 Academic Press. PMID- 11056072 TI - Training to Improve Calibration and Discrimination: The Effects of Performance and Environmental Feedback. AB - This study investigated whether calibration and discrimination are distinct or related aspects of probability judgment accuracy by examining the effects of two different training techniques. Participants received either performance feedback or environmental feedback, and we measured their improvement in calibration and discrimination as a function of feedback type. Whereas performance feedback reduced participants' overconfidence and environmental feedback improved discrimination, neither type of feedback led to an improvement on the other component. In fact, environmental feedback led to an increase in overconfidence. We take these results as evidence that calibration and discrimination are dissociable abilities that require separate training techniques for improvement. Copyright 2000 Academic Press. PMID- 11056073 TI - The Influence of Explanations for Selection Test Use, Outcome Favorability, and Self-Efficacy on Test-Taker Perceptions. AB - Previous research has shown that an explanation for an event can affect reactions to that event. This study examined this effect within a selection context by varying the type of explanation (causal, ideological, referential) and outcome favorability. A sample of 202 undergraduates completed a selection test and reported their perceptions of that test both before and after the outcome was known. Both the type of explanation and outcome favorability interacted with participants' self-efficacy in determining perceptions of validity and fairness. Implications and directions for future research are discussed. Copyright 2000 Academic Press. PMID- 11056074 TI - Evaluations of Outcome Sequences. AB - We extend the literature on intertemporal choice by investigating how possession of knowledge related to the present value of future outcomes (PV knowledge) affects the extent to which individuals weight certain attributes when evaluating outcome sequences. While PV-knowledgeable individuals can ascribe value to attributes according to their PV relevance (or irrelevance), unknowledgeable individuals cannot do so. Such knowledge, therefore, likely interacts with outcome-sequence attributes to affect the extent to which individuals exhibit impatience when evaluating outcome sequences. The main experimental findings indicate that higher PV knowledge increases the extent to which individuals value impatience (as opposed to improvement). However, these findings also reinforce a need to distinguish among impatience, improvement, and PV because some higher PV knowledge participants willingly sacrifice PV while exhibiting impatience (while others do so in order to gain improvement). Overall, PV considerations appear central, but not determinative, in higher PV-knowledgeable individuals' evaluations of outcome sequences. Copyright 2000 Academic Press. PMID- 11056075 TI - Judging Risk and Return of Financial Assets. AB - This article examines the relationship between judgments of risk and judgments of expected return of financial assets. It suggests that for unfamiliar assets, both risk and return judgments are derived from global preference toward the asset, whereas for familiar assets, these judgments tend to be derived from the ecological values of the asset's risk and expected return-their values in the financial markets. In addition, the article examines the role of causal schemas and the role of risk attitudes in mediating the relationships between judgments of risk and return of familiar and unfamiliar assets. Conceptual and practical questions concerning the nature, the meaning, and the assessment of risk and expected return are discussed. Copyright 2000 Academic Press. PMID- 11056076 TI - Treatment of epilepsy: existing therapies and future developments. AB - Epilepsy is a major public health issue, not least because of the aging population in many developed nations and the known increase in the frequency of epilepsy and seizures in later life. Despite the massive scale of the problem and much research, epilepsy remains poorly understood. Despite more than 20 approved drugs in the developed nations and several non-pharmacological options, up to 30% of patients are still refractory to treatment. Despite over a century of pharmacotherapy and neuroscience research, rational design of anti-epileptic drugs (AEDs) is only now starting to yield results, because of the heterogeneity of the disease and our still limited understanding of it. Discovery and development of AEDs has been especially difficult, because of the regulatory issues of satisfactorily proving safety and efficacy, ethical constraints on placebo-controlled trial designs, the fact that seizures are typically widely spaced in time, and the fact that the person undergoing the seizure is typically in no state to remember, let alone assess, what happened. Several non pharmacological therapies have been developed: brain surgery was first used more than a century ago; the ketogenic diet was first developed 80 years ago; and the vagus nerve stimulator was introduced recently. Pharmacotherapy remains the mainstay of treatment and is effective in most patients. AEDs can be roughly divided according to their time on the market. The first generation extends from the bromides and the barbiturates (the first of which was phenobarbital), to sodium valproate and carbamazepine. The second generation begins with felbamate and includes drugs approved from 1993 to 2000. "Next generation" drugs are still in clinical development and may reach the marketplace in the near future. Intensive research is being conducted both by pharmaceutical and biotech companies and by academic scientists and clinicians; our understanding of the condition is advancing rapidly but many challenges remain in discovering and developing better AEDs. PMID- 11056077 TI - Intracellular events and signaling pathways involved in sperm acquisition of fertilizing capacity and acrosome reaction. AB - Two processes, namely capacitation and acrosome reaction, are of fundamental importance in the fertilization of oocyte by spermatozoon. Physiologically occurring in the female genital tract, capacitation is a complex process, which renders the sperm cell capable for specific interaction with the oocyte. During capacitation, modification of membrane characteristics, enzyme activity and motility properties of spermatozoa render these cells able to penetrate oocyte investments and responsive to stimuli that induce acrosome reaction prior to fertilization. Physiological acrosome reaction occurs upon interaction of the spermatozoon with the zona pellucida protein ZP3. This is followed by liberation of several acrosomal enzymes and other constituents that facilitate penetration of the zona and expose molecules on the sperm equatorial segment that allows fusion of sperm membrane with the oolemma. The molecular mechanisms and the signal transduction pathways mediating the processes of capacitation and acrosome reaction have been partially defined, and appear to involve modifications of intracellular calcium and other ions, lipid transfer and phospholipid remodeling in sperm plasma membrane as well as changes in protein phosphorylation. Some of the kinases and phosphorylated proteins that are involved in the processes of capacitation and acrosome reaction have been now characterized. Characterization of sperm receptors to physiological inducers of acrosome reaction is in progress. This review summarizes the main signal transduction pathways involved in capacitation and acrosome reaction.Furthermore, the mechanisms underlying sperm DNA fragmentation are also briefly reviewed. PMID- 11056078 TI - Clinical applications of quinone-containing alkylating agents. AB - Quinone-containing alkylating agents are a class of chemical agents that have received considerable interest as anticancer drugs. These agents contain a quinone moiety that can be reduced and an alkylating group that can form covalent bonds with a variety of cellular components. The oxidation state of the quinone element can modulate the activity of the alkylating element, and reduction of the quinone is required for activation of the alkylating activity of many of these agents. The quinone element may also contribute to the cytotoxic activity of quinone-containing alkylating agents through the formation of reactive oxygen species during redox cycling. The natural product, mitomycin C, has been the most widely used quinone-containing alkylating agent in the clinic, but other quinone containing alkylating agents like porfiromycin, diaziquone, carbazilquinone, triaziquone and EO9 have also been used in the clinic for the treatment of cancer. In addition, many other quinone-containing alkylating agents have been tested in preclinical studies and the development of new agents is being actively pursued. This chapter describes the current and past clinical uses of these agents in the treatment of cancer and discusses new agents that are currently in clinical trials. PMID- 11056079 TI - The physiology and psychology of selective attention to touch. AB - In this chapter we review psychological and physiological experiments on selective attention to touch stimuli. We explore the role of selective attention in tactile target detection and search, determining those tasks that benefit from attention and those which can be effectively performed pre-attentively. We also try to determine the stage at which attentional selection occurs. We review electrophysiological and human brain imaging (PET, fMRI, MEG, SEP) studies to assess how early in the somatosensory processing pathway attentional modulation occurs. There is some evidence that the primary somatosensory cortex (S1) can show attentional effects. However, a number of studies have suggested that there is a hierarchy to attentional modulation in the somatosensory system, with the greatest effects being observed in secondary and association areas. PMID- 11056080 TI - Pathophysiologic role of selectins and their ligands in ischemia reperfusion injury. AB - Research findings are unveiling the potential role of leukocytes and leukocyte adhesion molecules such as selectins in ischemia-reperfusion injury (IRI). "Anti adhesion" therapy using selectin blocking agents may represent a new approach to treatment of the many diverse clinical disorders in which ischemia-reperfusion occurs, including transplantation, reperfusion after thrombotic events and shock. In this paper we review the pathophysiology of IRI, the different types of selectins and selectin ligands, the clinical implications of selectin blockade in different organs with IRI, and new insights into mechanism of action. PMID- 11056081 TI - Mechanisms of action of quinone-containing alkylating agents: DNA alkylation by aziridinylquinones. AB - Aziridinyl quinones can be activated by cellular reductases eg. DT-diaphorase and cytochrome P450 reductase to form highly reactive DNA alkylating agents. The mechanisms by which this activation and alkylation take place are many and varied. Using clinically relevant and experimental agents this review will describe many of these mechanisms. The agents discussed are Mitomycin C, EO9 and analogues, diaziridinylbenzoquinones and the pyrrolo[1, 2 alpha]benzimidazolequinones. PMID- 11056082 TI - High-throughput radioassays for autoantibodies to recombinant autoantigens. AB - Advances in immunology, biochemistry, and molecular biology have combined to allow the development of a large series of autoantibody assays utilizing recombinantly produced autoantigens. Labeled target proteins can be readily produced by in vitro transcription and translation of relevant cloned cDNA. The assays are carried out in the fluid phase and for most assays are more specific and sensitive than ELISA based assays. For some antigens (e.g. insulin) though ELISA assays detect antibodies following immunization, workshops indicate they are almost worthless for the diagnosis and prediction of type 1A diabetes. This new generation of radioassays is usually carried out in 96-well microtiter filtration plates that allow high throughput. Given such assays, individuals at high risk for type 1A diabetes, celiac disease, and Addison's disease can now be readily identified. PMID- 11056083 TI - Immune regulation by CD40-CD40-l interactions - 2; Y2K update. AB - CD40 is a cell surface receptor, which belongs to the TNF-R family, and which was first identified and functionally characterized on B lymphocytes. However, in recent years it has become clear that CD40 is expressed much broader, including expression on monocytes, dendritic cells, endothelial cells and epithelial cells. Therefore it is now thought that CD40 plays a more general role in immune regulation. The present paper reviews recent developments in this field of research, with main emphasis on CD40 signal transduction and on in vivo functions of CD40/CD40-L interactions. PMID- 11056084 TI - Electric activity spreads in the colonic longitudinal but not the circular musculature. Role in colonic motility. AB - The colon has a circular and longitudinal muscle layers and possesses an electric activity. The current study investigated the spread of this electric activity in the colonic musculature. The tests were performed in 16 patients (mean age 40.6 +/- 10.8 SD years, 10 men, 6 women) during operative correction of their incisional hernias in the anterior abdominal wall. Three electrodes were applied to the longitudinal muscle of the descending colon, and 3 to the circular muscle. The colonic pressure was monitored by a 10-F tube inserted per anus and connected to a pressure transducer. Electric waves in the form of pacesetter potentials (PPs) and action potentials (APs) were recorded from the longitudinal but not the circular muscle coat. The PPs and APs had the same frequency, amplitude and conduction velocity from the 3 electrodes of the same patient. The APs were associated with a rise of the intracolonic pressure. The results were reproducible in the individual subject. Being of the visceral type, the colonic smooth muscle fibers are controlled by non-nervous stimuli. Colonic motility is suggested to be effected by means of 2 mechanisms: the longitudinal muscle fibers activity through the action of the electric waves and the circular muscle fibers through the stretch reflex. PMID- 11056085 TI - Nitric oxide and viral cardiomyopathy. PMID- 11056086 TI - Direct proinflammatory effect of C-reactive protein on human endothelial cells. AB - BACKGROUND: The acute-phase reactant C-reactive protein (CRP) is an important risk factor for coronary heart disease. However, the possible effects of CRP on vascular cells are not known. METHODS AND RESULTS: We tested the effects of CRP on expression of adhesion molecules in both human umbilical vein and coronary artery endothelial cells. Expression of vascular cell adhesion molecule (VCAM-1), intercellular adhesion molecule (ICAM-1), and E-selectin was assessed by flow cytometry. Incubation with recombinant human CRP (10 microg/mL) for 24 hours induced an approximately 10-fold increase in expression of ICAM-1 and a significant expression of VCAM-1, whereas a 6-hour incubation induced significant E-selectin expression. Adhesion molecule induction was similar to that observed in endothelial cells activated with interleukin-1beta. In coronary artery endothelial cells, induction of ICAM-1 and VCAM-1 was already present at 5 microg/mL and reached a maximum at 50 microg/mL, at which point a substantial increase in expression of E-selectin was also evident. The CRP effect was dependent on presence of human serum in the culture medium, because no effect was seen in cells cultured with serum-free medium. In contrast, interleukin-1beta was able to induce adhesion molecule expression in the absence of human serum. CONCLUSIONS: CRP induces adhesion molecule expression in human endothelial cells in the presence of serum. These findings support the hypothesis that CRP may play a direct role in promoting the inflammatory component of atherosclerosis and present a potential target for the treatment of atherosclerosis. PMID- 11056087 TI - Olive phenol hydroxytyrosol prevents passive smoking-induced oxidative stress. AB - BACKGROUND: Oxidative stress is involved in the onset of several degenerative disorders, and epidemiological studies indicate that a high intake of dietary antioxidants, as in the case of the Mediterranean basin, is protective. Olive mill waste waters (OMWWs) are a byproduct of olive oil production rich in phenolic antioxidants, such as hydroxytyrosol. We tested the effects of a low dose of an OMWW extract in a model of sidestream smoke-induced oxidative stress in rats by evaluating the urinary excretion of 8-iso-prostaglandin (PG) F(2alpha) (iPF(2alpha)-III). METHODS AND RESULTS: An OMWW extract (5 mg/kg, providing 414 microg/kg of hydroxytyrosol) was administered to rats daily for 4 days, during which time the animals were exposed to sidestream smoke for 20 minutes once a day. Daily urines were collected, and the urinary excretion of 8-iso-PGF(2alpha) was evaluated as an index of oxidative stress-induced in vivo lipid peroxidation. The exposure of rats to passive smoking increased the urinary excretion of 8-iso PGF(2alpha) by 44+/-4.2% at 48 hours and by 55+/-10% at 96 hours. Treatment with the OMWW extract was able to completely prevent the increase at 48 hours and resulted in lower increments (34+/-18% versus 55+/-10%) of 8-iso-PGF(2alpha) excretion at 96 hours. CONCLUSIONS: A low dose of hydroxytyrosol, administered through OMWW, reduces the consequences of sidestream smoke-induced oxidative stress in rats. PMID- 11056088 TI - Endothelial dysfunction of coronary resistance arteries is improved by tetrahydrobiopterin in atherosclerosis. AB - BACKGROUND: Tetrahydrobiopterin (BH4), an essential cofactor for the synthesis of NO, improves endothelial dysfunction after ischemia/reperfusion. Therefore, we hypothesized that reduction of BH4 is involved in the attenuation of endothelium dependent vasodilation in atherosclerosis, and we investigated the effect of alterations of the BH4 level on the vasodilatory potential of coronary resistance vessels from humans and pigs with atherosclerosis. METHODS AND RESULTS: Coronary arterioles were obtained from patients undergoing CABG (atherosclerosis group) or valve replacement (control group) and from pigs fed either a standard diet (control group) or atherogenic diet (atherosclerosis group). After isolation, vessels were cannulated, pressurized, and placed on the stage of an inverted microscope. Dose-response curves were investigated in response to the endothelium dependent agonists histamine, serotonin, and acetylcholine (for pigs, substance P) and to the endothelium-independent agonist sodium nitroprusside (SNP) under control conditions and before and after incubation of the vessels with sepiapterin (substrate for BH4 synthesis). In vessels from patients and from animals with atherosclerosis, compared with vessels from the control groups, there was a significant (P:<0.05) reduction of vasodilation to all tested endothelium-dependent agonists but not to SNP. After application of sepiapterin, the responses to the endothelium-dependent agonists but not to SNP were significantly improved in vessels from the atherosclerosis groups. Sepiapterin did not influence vascular reactivity in the control groups. CONCLUSIONS: Atherosclerosis severely compromises endothelial function of coronary resistance arteries. Administration of sepiapterin leads to a significant improvement of endothelium-dependent vasodilatation to different agonists in vessels from humans and pigs with atherosclerosis. Therefore, we conclude that a reduced availability of BH4 is involved in the development of endothelial dysfunction in atherosclerosis. PMID- 11056089 TI - Coronary composition and macrophage infiltration in atherectomy specimens from patients with diabetes mellitus. AB - BACKGROUND: Lipid-rich, inflamed atherosclerotic lesions are associated with plaque rupture and thrombosis, which are the most important causes of death in patients with diabetes mellitus. This study was designed to quantify lipid composition and macrophage infiltration in the coronary lesions of patients with diabetes mellitus. METHODS AND RESULTS: A total of 47 coronary atherectomy specimens from patients with diabetes mellitus were examined and compared with 48 atherectomy specimens from patients without diabetes. Plaque composition was characterized by trichrome staining. Macrophage infiltration was characterized by immunostaining. Clinical and demographic data were similar in both groups. The percentage of total area occupied by lipid-rich atheroma was larger in specimens from patients with diabetes (7+/-2%) than in specimens from patients without diabetes (2+/-1%; P:=0.01), and the percentage of total area occupied by macrophages was larger in specimens from patients with diabetes (22+/-3%) than in specimens from patients without diabetes (12+/-1%; P:=0.003). The incidence of thrombus was also higher in specimens from patients with diabetes than in specimens from patients without diabetes (62% versus 40%; P:=0.04). Plaque composition, macrophage infiltration, and thrombus were similar in lesions from diabetic patients treated with insulin compared with lesions from patients treated with sulfonylureas or diet. CONCLUSIONS: Coronary tissue from patients with diabetes exhibits a larger content of lipid-rich atheroma, macrophage infiltration, and subsequent thrombosis than tissue from patients without diabetes. These differences suggest an increased vulnerability for coronary thrombosis in patients with diabetes mellitus. PMID- 11056090 TI - Overexpression of eotaxin and the CCR3 receptor in human atherosclerosis: using genomic technology to identify a potential novel pathway of vascular inflammation. AB - BACKGROUND: Unstable atherosclerotic lesions typically have an abundant inflammatory cell infiltrate, including activated T cells, macrophages, and mast cells, which may decrease plaque stability. The pathophysiology of inflammatory cell recruitment and activation in the human atheroma is incompletely described. METHODS AND RESULTS: We hypothesized that differential gene expression with DNA microarray technology would identify new genes that may participate in vascular inflammation. RNA isolated from cultured human aortic smooth muscle cells treated with tumor necrosis factor-alpha (TNF-alpha) was examined with a DNA microarray with 8600 genes. This experiment and subsequent Northern analyses demonstrated marked increases in steady-state eotaxin mRNA (>20 fold), a chemokine initially described as a chemotactic factor for eosinophils. Because eosinophils are rarely present in human atherosclerosis, we then studied tissue samples from 7 normal and 14 atherosclerotic arteries. Immunohistochemical analysis demonstrated overexpression of eotaxin protein and its receptor, CCR3, in the human atheroma, with negligible expression in normal vessels. Eotaxin was predominantly located in smooth muscle cells. The CCR3 receptor was localized primarily to macrophage rich regions as defined by immunopositivity for CD 68; a minority of mast cells also demonstrated immunopositivity for the CCR3 receptor. CONCLUSIONS: Eotaxin and its receptor, CCR3, are overexpressed in human atherosclerosis, suggesting that eotaxin participates in vascular inflammation. These data demonstrate how genomic differential expression technology can identify novel genes that may participate in the stability of atherosclerotic lesions. PMID- 11056091 TI - Bradykinin stimulates tissue plasminogen activator release from human forearm vasculature through B(2) receptor-dependent, NO synthase-independent, and cyclooxygenase-independent pathway. AB - BACKGROUND: Bradykinin stimulates dose-dependent tissue plasminogen activator (tPA) release from human endothelium. Although bradykinin is known to cause vasodilation through B(2) receptor-dependent effects on NO, prostacyclin, and endothelium-derived hyperpolarizing factor production, the mechanism(s) underlying tPA release is unknown. METHODS AND RESULTS: We measured the effects of intra-arterial bradykinin (100, 200, and 400 ng/min), acetylcholine (15, 30, and 60 microg/min), and nitroprusside (0.8, 1.6, and 3.2 microg/min) on forearm vasodilation and tPA release in healthy volunteers in the presence and absence of (1) the B(2) receptor antagonist HOE 140 (100 microg/kg IV), (2) the NO synthase inhibitor L-N:(G)-monomethyl-L-arginine (L-NMMA, 4 micromol/min intra arterially), and (3) the cyclooxygenase inhibitor indomethacin (50 mg PO TID). B(2) receptor antagonism attenuated vasodilator (P:=0.004) and tPA (P:=0.043) responses to bradykinin, without attenuating the vasodilator response to nitroprusside (P:=0.36). L-NMMA decreased basal forearm blood flow (from 2.35+/ 0.31 to 1. 73+/-0.22 mL/min per 100 mL, P:=0.01) and blunted the vasodilator response to acetylcholine (P:=0.013) and bradykinin (P:=0.07, P:=0. 038 for forearm vascular resistance) but not that to nitroprusside (P:=0.47). However, there was no effect of L-NMMA on basal (P:=0.7) or bradykinin-stimulated tPA release (P:=0.45). Indomethacin decreased urinary excretion of the prostacyclin metabolite 2, 3-dinor-6-keto-prostaglandin F(1alpha) (P:=0.04). The vasodilator response to endothelium-dependent (P:=0.019 for bradykinin) and endothelium independent (P:=0.019) vasodilators was enhanced during indomethacin administration. In contrast, there was no effect of indomethacin alone (P:=0.99) or indomethacin plus L-NMMA (P:=0.36) on bradykinin-stimulated tPA release. CONCLUSIONS: These data indicate that bradykinin stimulates tPA release from human endothelium through a B(2) receptor-dependent, NO synthase-independent, and cyclooxygenase-independent pathway. Bradykinin-stimulated tPA release may represent a marker for the endothelial effects of endothelium-derived hyperpolarizing factor. PMID- 11056092 TI - Common cholesteryl ester transfer protein mutations, decreased HDL cholesterol, and possible decreased risk of ischemic heart disease: The Copenhagen City Heart Study. AB - BACKGROUND: Cholesteryl ester transfer protein (CETP) mediates the transfer of cholesteryl ester from HDL in exchange for triglycerides in apolipoprotein B containing lipoproteins. METHODS AND RESULTS: We studied 2 common mutations in CETP, A373P and R451Q, in 8467 healthy women and men from the Danish general population and in 1636 Danish women and men with ischemic heart disease. The prevalence of 373P and 451Q was 0.10 and 0.07, respectively, for heterozygous carriers and 0.003 and 0.002, respectively, for homozygous carriers. All carriers of the 451Q allele also carried the 373P allele. HDL cholesterol in female noncarriers, heterozygotes, and homozygotes of 373P was 1.74+/-0.01 (mean+/-SE), 1.62+/-0.02, and 1.38+/-0.09 mmol/L, respectively (ANOVA, P:<0.001). In men, equivalent values were 1.40+/-0.01, 1.26+/-0.02, and 1.19+/-0.09 mmol/L, respectively (ANOVA, P:<0.001). HDL cholesterol decreased similarly as a function of 451Q genotypes and all 373P/451Q genotype combinations. Furthermore, apolipoprotein AI and the HDL cholesterol/apolipoprotein AI ratio was also lower in carriers of either of these mutations for both sexes. Finally, the CETP genotype was not associated with risk of ischemic heart disease unless we adjusted for HDL cholesterol: female heterozygous and homozygous carriers versus noncarriers had 36% lower risk of ischemic heart disease (95% CI 4% to 57%); in male carriers, we observed a similar trend. CONCLUSIONS: The A373P/R451Q polymorphism in CETP is associated with decreases in HDL cholesterol of 0.12 to 0.36 mmol/L in women and 0.14 to 0.21 mmol/L in men and possibly with a paradoxical 36% decrease in the risk of ischemic heart disease in women. PMID- 11056093 TI - Change in level of physical activity and risk of all-cause mortality or reinfarction: The Corpus Christi Heart Project. AB - BACKGROUND: The role of physical activity (PA) in reducing the risk of all-cause mortality or reinfarction after a first myocardial infarction (MI) remains unresolved, particularly for minority populations. The association between change in level of PA and risk of death or reinfarction was studied in 406 Mexican American and non-Hispanic white women and men who survived a first MI. METHODS AND RESULTS: MI patients were interviewed at baseline and annually thereafter about PA, medical history, and risk factors of coronary heart disease. Change in level of PA after the index MI was categorized as (1) sedentary, no change (referent group), (2) decreased activity, (3) increased activity, and (4) active, no change. Over a 7-year period, the relative risk (95% CI) of death was as follows: 0.21 (0.10 to 0.44) for the active, no change group; 0.11 (0.03 to 0.46) for the increased activity group; and 0.49 (0.26 to 0.90) for the decreased activity group. The relative risk of reinfarction was as follows: 0.40 (0.24 to 0.66) for the active, no change group; 0.22 (0.09 to 0.50) for the increased activity group; and 0.93 (0.59 to 1.42) for the decreased activity group. CONCLUSIONS: These findings are consistent with a beneficial role of PA for Mexican American and non-Hispanic white women and men who survive a first MI and have practical implications for the management of MI survivors. PMID- 11056094 TI - Angiotensin II receptor subtypes in the skeletal muscle vasculature of patients with severe congestive heart failure. AB - BACKGROUND: Vascular remodeling occurs in the skeletal muscle of patients with severe congestive heart failure (CHF); this remodeling is mediated in part by increased activity of the renin-angiotensin system. Animal models suggest that in the vasculature, angiotensin II receptor type 2 (AT2-R) expression may be upregulated in pathological states associated with vascular remodeling. The therapeutic effects of an AT1-R antagonist may, therefore, be in part due to increased plasma angiotensin II levels, which stimulate AT2-R. However, whether AT2-R is expressed in the skeletal muscle vasculature of patients with severe CHF is unknown. METHODS AND RESULTS: The steady-state transcript levels of the AT1-R and AT2-R genes were analyzed by reverse transcription-polymerase chain reaction in RNA samples prepared from the skeletal muscle of 12 patients with severe CHF (f1.gif" BORDER="0">O(2)<10 mL. kg(-1). min(-1)) and 5 age-matched healthy subjects who underwent vastus lateralis biopsies. Human fetal skeletal muscle RNA served as a positive control for the expression of AT1-R and AT2-R gene transcripts. Transcripts from the AT1-R gene were detected readily in all samples. In contrast, transcripts from the AT2-R gene were only detected in fetal skeletal muscle samples and could not be detected in the skeletal muscle vasculature of healthy subjects or that of CHF patients, who were treated with either angiotensin-converting enzyme inhibitors or AT1-R antagonists. CONCLUSIONS: The AT2-R gene is not expressed in the skeletal muscle of patients with CHF. In the absence of detectable AT2-R gene transcripts, the AT2-R pathway is unlikely to contribute to the effects of AT1-R antagonists on the skeletal muscle vasculature in patients with severe CHF. PMID- 11056095 TI - Quantitative general theory for periodic breathing in chronic heart failure and its clinical implications. AB - BACKGROUND: In patients with chronic heart failure (CHF), periodic breathing (PB) predicts poor prognosis. Clinical studies have identified numerous risk factors for PB (which also includes Cheyne-Stokes respiration). Computer simulations have shown that oscillations can arise from delayed negative feedback. However, no simple general theory quantitatively explains PB and its mechanisms of treatment using widely-understood clinical concepts. Therefore, we introduce a new approach to the quantitative analysis of the dynamic physiology governing cardiorespiratory stability in CHF. METHODS AND RESULTS: An algebraic formula was derived (presented as a simple 2D plot), enabling prediction from easily acquired clinical data to determine whether respiration will be unstable. Clinical validation was performed in 20 patients with CHF (10 with PB and 10 without) and 10 healthy normal subjects. Measurements, including chemoreflex sensitivity (S) and delay (delta), alveolar volume (V(L)), and end-tidal CO(2) fraction (C), were applied to the stability formula. The breathing pattern was correctly predicted in 28 of the 30 subjects. The principal combined parameter (CS)x(delta/V(L)) was higher in patients with PB (14.2+/-3.0) than in those without PB (3.1+/-0.5; P:=0.0005) or in normal controls (2.4+/-0.5; P:=0.0003). This was because of differences in both chemoreflex sensitivity (1749+/-235 versus 620+/-103 and 526+/-104 L/min per atm CO(2); P:=0.0001 and P:<0.0001, respectively) and chemoreflex delay (0.53+/-0.06 vs 0.40+/-0.06 and 0.30+/-0.04 min; P:=NS and P:=0.02). CONCLUSION: This analytical approach identifies the physiological abnormalities that are important in the genesis of PB and explicitly defines the region of predicted instability. The clinical data identify chemoreflex gain and delay time (rather than hyperventilation or hypocapnia) as causes of PB. PMID- 11056096 TI - Acute hemodynamic and clinical effects of levosimendan in patients with severe heart failure. Study Investigators. AB - BACKGROUND: We determined the short-term hemodynamic and clinical effects of levosimendan, a novel calcium-sensitizing agent, in patients with decompensated heart failure. METHODS AND RESULTS: One hundred forty-six patients with New York Heart Association functional class III or IV heart failure (mean left ventricular ejection fraction 21+/-1%) who had a pulmonary capillary wedge pressure >/=15 mm Hg and a cardiac index /=27 kg x m(-2), n=16) or hypertensive patients (blood pressure >140/90 mm Hg, n=21) (50+/-8 and 55+/-10 g x m(-2.7), respectively) than in 13 nonobese, normotensive subjects (40+/-8 g x m(-2.7), P:=0.0004). In a multivariate model adjusting for sex, age, BMI, and blood pressure, neither insulin concentrations (fasting or postglucose) nor insulin sensitivity or secretory rates were significant correlates of LVM. Systolic blood pressure (P:=0.003) and BMI (P:=0.01) were the only independent correlates of LVM. From the regression, the impact of hypertension (as a systolic pressure of 180 versus 140 mm Hg=+20%) was twice as large as that of obesity (as a BMI of 35 versus 25 kg x m(-2)=+11%), the two factors being additive. CONCLUSIONS: When adequate account is taken of body mass and blood pressure, insulin, as concentration, secretion, or action, is not an independent determinant of LVM in nondiabetic subjects. PMID- 11056099 TI - Effect of second-phase duration on the strength-duration relation for human transvenous defibrillation. AB - BACKGROUND: The mechanism by which biphasic waveforms improve defibrillation efficacy is unclear. In addition, the optimal shape of the biphasic waveforms remains controversial. Animal experiments suggest that prolonging the duration of the second phase longer than the first worsens defibrillation thresholds (DFT). The purpose of this study was to determine the strength-duration relation for the second phase of a biphasic defibrillation waveform in humans. METHODS AND RESULTS: This was a prospective, randomized study of biphasic DFT in 36 patients; a uniform dual-coil transvenous lead system was used. In each patient, 3 DFTs were determined with the pulse duration for the second phase of the defibrillation waveform varying between 1 and 18 ms. The duration of the first phase was fixed at 6 ms and the capacitance was 150 microF. There was a significant increase in the leading edge voltage at DFT only when the second phase pulse duration was decreased to 1 ms. There was no increase in DFT voltage even when the second-phase pulse duration was increased from 2 to 18 ms. Similar relations were observed for stored energy, leading edge current, or phase 2 energy. The normalized average current delivered during phase 2 decreased monotonically with increasing phase 2 duration. CONCLUSIONS: In humans, the biphasic DFT voltage or energy is increased only when the second phase of the waveform is <2 ms. The DFT voltage is insensitive to increasing the second phase of the defibrillator waveform to as long as 18 ms, or 3 times the duration of the first phase of the waveform. PMID- 11056100 TI - Role of monocyte chemoattractant protein-1 in cardiovascular remodeling induced by chronic blockade of nitric oxide synthesis. AB - BACKGROUND: Chronic inhibition of endothelial nitric oxide (NO) synthesis by the administration of N:(omega)-nitro-L-arginine methyl ester (L-NAME) to rats induces early vascular inflammatory changes (monocyte infiltration into coronary vessels and monocyte chemoattractant protein-1 [MCP-1] expression) as well as subsequent arteriosclerosis (medial thickening and perivascular fibrosis) and cardiac fibrosis. However, the role of MCP-1 in this process is not known. METHODS AND RESULTS: We investigated the effect of a specific monoclonal anti-MCP 1 neutralizing antibody in rats treated with L-NAME to determine the role of monocytes in the regulation of cardiovascular remodeling. We found increased expression of MCP-1 mRNA in vascular endothelial cells and monocytes in inflammatory lesions. Cotreatment with an anti-MCP-1 antibody, but not with control IgG, prevented the L-NAME-induced early inflammation and reduced late coronary vascular medial thickening. In contrast, the anti-MCP-1 antibody did not decrease the development of perivascular fibrosis, the expression of transforming growth factor (TGF)-beta(1) mRNA, or systolic pressure overload induced by L-NAME administration. CONCLUSIONS: These results suggest that MCP-1 is necessary for the development of medial thickening as well as monocyte recruitment. In contrast, the pathogenesis of fibrosis may involve other factors, such as TGF beta(1). PMID- 11056101 TI - Chronic alpha-adrenergic receptor stimulation modulates the contractile phenotype of cardiac myocytes in vitro. AB - BACKGROUND: Heart failure is characterized by contractile dysfunction of the myocardium and elevated sympathetic activity. We tested the hypothesis that chronic alpha-adrenergic (alpha-ADR) stimulation modifies the molecular and contractile phenotype of cardiac myocytes. METHODS AND RESULTS: Adult rat ventricular myocytes in culture were exposed to alpha-ADR stimulation (norepinephrine + propranolol) for 48 hours. alpha-ADR stimulation decreased the mRNAs for sarcoplasmic reticulum Ca(2+)-ATPase and Ca(2+) release channel by 56% and 52%, respectively, and increased mRNA and protein for the Na(+)-Ca(2+) exchanger by 70% and 39%, respectively. After washout of the alpha-ADR agonist, simultaneous measurement of [Ca(2+)](i) transients with fura 2 and myocyte shortening by video edge-detection showed that [Ca(2+)](i) amplitude and myocyte shortening were decreased in alpha-ADR-treated myocytes, and the time to peak and time from peak to 80% decline of both [Ca(2+)](i) and myocyte shortening were increased. The concentration-response curve for myocyte shortening by the Na(+) channel activator veratridine was shifted leftward in alpha-ADR-stimulated myocytes (EC(50), 21.6+/-4.6 versus 105.8+/-10.5 nmol/L, P:<0.001). CONCLUSIONS: Chronic alpha-ADR stimulation of cardiac myocytes causes decreases in the expression of sarcoplasmic reticulum Ca(2+)-ATPase and the Ca(2+) release channel that are associated with decreases in [Ca(2+)](i) and contractility. alpha-ADR stimulation simultaneously increases Na(+)-Ca(2+) exchanger expression, thereby increasing sensitivity to intracellular Na(+). PMID- 11056102 TI - Angiogenesis is induced in a rabbit model of hindlimb ischemia by naked DNA encoding an HIF-1alpha/VP16 hybrid transcription factor. AB - BACKGROUND: Hypoxia-inducible factor-1 (HIF-1) is a heterodimeric transcription factor that regulates expression of genes involved in O(2) homeostasis, including vascular endothelial growth factor (VEGF), a potent stimulator of angiogenesis. We sought to exploit this native adaptive response to hypoxia as a treatment for chronic ischemia. METHODS AND RESULTS: A hybrid protein consisting of DNA-binding and dimerization domains from the HIF-1alpha subunit and the transactivation domain from herpes simplex virus VP16 protein was constructed to create a strong, constitutive transcriptional activator. After transfection into HeLa, C6, and Hep3B cells, this chimeric transcription factor was shown to activate expression of the endogenous VEGF gene, as well as several other HIF-1 target genes in vitro. The bioactivity of HIF-1alpha/VP16 hybrid gene transfer in vivo was examined in a rabbit model of hindlimb ischemia. Administration of HIF 1alpha/VP16 was associated with significant improvements in calf blood pressure ratio, angiographic score, resting and maximal regional blood flow, and capillary density (all P:<0.01). CONCLUSIONS: The HIF-1alpha/VP16 hybrid transcription factor is able to promote significant improvement in perfusion of an ischemic limb. These results confirm the feasibility of a novel approach for therapeutic angiogenesis in which neovascularization may be achieved indirectly by use of a transcriptional regulatory strategy. PMID- 11056103 TI - Intravascular adenovirus-mediated VEGF-C gene transfer reduces neointima formation in balloon-denuded rabbit aorta. AB - BACKGROUND: Gene transfer to the vessel wall may provide new possibilities for the treatment of vascular disorders, such as postangioplasty restenosis. In this study, we analyzed the effects of adenovirus-mediated vascular endothelial growth factor (VEGF)-C gene transfer on neointima formation after endothelial denudation in rabbits. For comparison, a second group was treated with VEGF-A adenovirus and a third group with lacZ adenovirus. Clinical-grade adenoviruses were used for the study. METHODS AND RESULTS: Aortas of cholesterol-fed New Zealand White rabbits were balloon-denuded, and gene transfer was performed 3 days later. Animals were euthanized 2 and 4 weeks after the gene transfer, and intima/media ratio (I/M), histology, and cell proliferation were analyzed. Two weeks after the gene transfer, I/M in the lacZ-transfected control group was 0. 57+/-0.04. VEGF-C gene transfer reduced I/M to 0.38+/-0.02 (P:<0.05 versus lacZ group). I/M in VEGF-A treated animals was 0.49+/-0.17 (P:=NS). The tendency that both VEGF groups had smaller I/M persisted at the 4-week time point, when the lacZ group had an I/M of 0.73+/-0.16, the VEGF-C group 0.44+/-0.14, and the VEGF-A group 0. 63+/-0.21 (P:=NS). Expression of VEGF receptors 1, 2, and 3 was detected in the vessel wall by immunocytochemistry and in situ hybridization. As an additional control, the effect of adenovirus on cell proliferation was analyzed by performing gene transfer to intact aorta without endothelial denudation. No differences were seen in smooth muscle cell proliferation or I/M between lacZ adenovirus and 0.9% saline-treated animals. CONCLUSIONS: Adenovirus-mediated VEGF-C gene transfer may be useful for the treatment of postangioplasty restenosis and vessel wall thickening after vascular manipulations. PMID- 11056104 TI - Calcineurin inhibitor attenuates left ventricular hypertrophy, leading to prevention of heart failure in hypertensive rats. AB - BACKGROUND: There is controversy regarding the contribution of calcineurin activation to the development of pressure-overload left ventricular (LV) hypertrophy and heart failure. The aim of this study was to explore whether the inhibition of calcineurin may prevent the transition to heart failure in hypertensive rats and, if so, to clarify in which developmental stage of LV hypertrophy calcineurin plays a key role. METHODS AND RESULTS: Dahl salt sensitive rats placed on an 8% NaCl diet from the age of 7 weeks (hypertensive rats) were randomized to no treatment (n=6) or treatment with the calcineurin inhibitor FK506 (1 mg x kg(-1) x d(-1)) from 8 weeks (FKE, n=7) or from 17 weeks (FKL, n=7). Rats placed on a 0.3% NaCl diet were defined as control rats (n=6). The administration of FK506 from 8 weeks attenuated, although it did not block, LV hypertrophy observed in the untreated rats and prevented the transition to heart failure. The development of LV fibrosis, however, was not attenuated by the administration of FK506 from 8 weeks. The administration of FK506 from 17 weeks brought no benefit for cardiac remodeling or LV function and failed to prevent heart failure. CONCLUSIONS: Calcineurin inhibition, if started from the initial stage of pressure overload, attenuated the development of LV hypertrophy without any effect on LV fibrosis and prevented the transition to heart failure. The activation of calcineurin is involved in the development of LV hypertrophy but not of LV fibrosis, and this involvement may be crucial at the initial stage. PMID- 11056105 TI - Nitric oxide inhibits dystrophin proteolysis by coxsackieviral protease 2A through S-nitrosylation: A protective mechanism against enteroviral cardiomyopathy. AB - BACKGROUND: Infection with enteroviruses like coxsackievirus B3 (CVB3) as well as genetic dystrophin deficiency can cause dilated cardiomyopathy. We recently identified cleavage and functional impairment of dystrophin by the viral protease 2A during CVB3-infection as a molecular mechanism that may contribute to the pathogenesis of enterovirus-induced cardiomyopathy. Nitric oxide (NO) is elevated in human dilated cardiomyopathy, but the relevance of this finding is unknown. In mice, NO inhibits CVB3 myocarditis. Therefore, we investigated the effects of NO on the coxsackieviral protease 2A. METHODS AND RESULTS: In vitro, NO donors like PAPA-NONOate inhibited the cleavage of human and mouse dystrophin by recombinant coxsackievirus B protease 2A in a dose-dependent manner (IC(50), 51 micromol/L). In CVB3-infected HeLa cells, addition of the NO donor SNAP inhibited protease 2A catalytic activity on dystrophin. Because this inhibitory effect was reversed by the thiol-protecting agent DTT, we investigated whether NO S:-nitrosylates the protease 2A. In vitro, NO nitrosylated the active-site cysteine (C110) of the coxsackieviral protease 2A, as demonstrated by site-directed mutagenesis. Within living COS-7 cells, SNAP-induced S:-nitrosylation of this site was confirmed with electron spin resonance spectroscopy. CONCLUSIONS: These data demonstrate inactivation of a coxsackieviral protease 2A by NO through active-cysteine S: nitrosylation in vitro and intracellularly. Given that the enteroviral protease 2A cleaves mouse and human dystrophin, NO may be protective in human heart failure with an underlying enteroviral pathogenesis through inhibition of dystrophin proteolysis. PMID- 11056106 TI - Recognition of left atrial aneurysm by fetal echocardiography. PMID- 11056107 TI - AHA Dietary Guidelines: revision 2000: A statement for healthcare professionals from the Nutrition Committee of the American Heart Association. PMID- 11056108 TI - Recommendations for the management of patients with unruptured intracranial aneurysms: A statement for healthcare professionals from the Stroke Council of the American Heart Association. PMID- 11056109 TI - American College of Cardiology/American Heart Association Clinical Competence Statement on invasive electrophysiology studies, catheter ablation, and cardioversion: A report of the American College of Cardiology/American Heart Association/American College of Physicians-American Society of Internal Medicine Task Force on Clinical Competence. PMID- 11056110 TI - Percutaneous coil embolization of multiple arteriovenous malformations in left lung causing persistent hypoxia. PMID- 11056111 TI - Regular aerobic exercise augments endothelium-dependent vascular relaxation in normotensive and hypertensive subjects: role of endothelium-derived nitric oxide. PMID- 11056112 TI - Coronary aneurysm after endovascular brachytherapy: true or false? PMID- 11056113 TI - A knockout may not always be a knockout. PMID- 11056114 TI - Catheter-mediated linear block in the atria. PMID- 11056115 TI - Investigation of the infertile couple: investigation of the infertile couple in the era of assisted reproductive technology: a time for reappraisal. AB - At present, several of the elements in widespread use in basic infertility testing are in dispute, marked variability exists in the work-up among specialists, and practice patterns are influenced both by modern assisted reproductive technologies (ART) and the increasing age of couples seeking help for infertility. This article is intended to stimulate the debate on a possible (lack of) usefulness of conventional methods of infertility evaluation in relation to both the modern techniques of assisted reproduction and the woman's age. PMID- 11056116 TI - The role of LH in ovarian stimulation: exogenous LH: let's design the future. AB - Historically, follicular stimulation protocols have included both FSH and LH in an attempt to mimic the physiology of normal human folliculogenesis. However, many recent gonadotrophin administration regimens have completely eliminated LH bioactivity. The importance and the amount of LH necessary for optimal follicular stimulation has been a topic of debate. Several recent studies have added to our understanding of the actions of androgens, oestrogens, gonadotrophins, and insulin on the follicle-oocyte unit, allowing a less speculative approach. Moreover, the availability of human gonadotrophins synthesized by recombinant DNA technology and gonadotrophin-releasing hormone (GnRH) antagonists, should soon permit a precise in-vivo assessment and re-evaluation of the historical 2-cell, two-gonadotrophin hypothesis. These pharmacological tools may also provide essential insights into the physiological roles of FSH and LH in human follicular development and oocyte maturation. The recombinant gonadotrophins give clinicians the unique opportunity to tailor ovarian stimulation regimens according to the patient's medical history, in an effort both to maximize oocyte yield and to improve oocyte quality. PMID- 11056117 TI - Severe OHSS: yes, there is a strategy to prevent it! AB - Effective measures to prevent ovarian hyperstimulation syndrome (OHSS) remain controversial. It became almost 'common knowledge' that there is no strategy that may completely prevent OHSS. Extensive clinical experience (albeit not derived from prospective randomized studies) clearly documents the ability of a single administration of gonadotrophin-releasing hormone (GnRH) agonist to effectively trigger ovulation, while completely eliminating any threat of clinically significant OHSS. This strategy cannot be used if the pituitary is down-regulated (as is the case in most assisted reproductive cycles today), however, the newly introduced GnRH antagonists open new opportunities for implementing this strategy, since the pituitary preserves its responsiveness to GnRH agonists. Combining GnRH antagonist-based ovarian stimulation (particularly in 'high responders'), with GnRH agonist-driven ovulation triggering will make severe OHSS a disease of the past in assisted reproduction. PMID- 11056118 TI - Criteria predicting the absence of spermatozoa in the Sertoli cell-only syndrome can be used to improve success rates of sperm retrieval. AB - In patients with non-obstructive azoospermia, testicular sperm extraction (TESE) is a method of choice to recover spermatozoa as a male therapeutic approach in intracytoplasmic sperm injection (ICSI) programmes. However, the efficacy of TESE in this indication is burdened by a frequent failure of sperm recovery, which renders useless both the invasive testicular intervention and ovarian stimulation of the patient's spouse. One of the most frequent pathological pictures characterizing complete absence of spermatozoa is germinal aplasia (Sertoli cell- only syndrome or SCOS). Two different histological patterns of SCOS have been already described during the past five decades. These two patterns can be characterized as the congenital (pure) and the secondary (mixed) forms. Both patterns, with different prognosis to retrieve spermatozoa by therapeutic testicular biopsy, are frequently confused when TESE is performed during ICSI programmes. Useful criteria to predict the absence of spermatozoa can be obtained by a definite recognition of the two typical histological patterns during the diagnostic testicular biopsy. The diagnosis of congenital or acquired SCOS can be refined by endocrine, chemical, immunohistochemical and molecular biology aids. Reduction of both sperm retrieval failure and unnecessary ovarian stimulation can be achieved by combination of these methods. PMID- 11056119 TI - Microsurgical TESE and the distribution of spermatogenesis in non-obstructive azoospermia. AB - We wished to map the distribution of spermatogenesis in different regions of the testis in 58 men with non-obstructive azoospermia, and to develop a rational microsurgical strategy for the testicular sperm extraction (TESE) procedure. One goal was to maximize the chances for retrieving spermatozoa from such men, to minimize tissue loss and pain, and to preserve the chance for successful future procedures. Another goal was to expand upon the previously reported quantitative histological analysis of testicular tissue in 45 azoospermic men undergoing conventional TESE, this time using microsurgical as well as histological mapping. Tubular fullness observed at microsurgery and the presence of spermatozoa in the TESE specimen was compared with the quantitative histological analysis of spermatogenesis. Thus, our conclusions about the distribution of spermatogenesis are based on our experience with TESE in 103 consecutive cases of non-obstructive azoospermia. It was confirmed that men with non-obstructive azoospermia caused by germinal failure have a mean of 0 to 3 mature spermatids per seminiferous tubule in contrast to 17-35 mature spermatids per tubule in men with normal spermatogenesis and obstructive azoospermia. The former represented the threshold of quantitative spermatogenesis which must be exceeded in order for spermatozoa to 'spill over' into the ejaculate. Both testicular 'mapping' by multiple biopsy (n = 15) and microsurgical removal of contiguous strips of testicular tissue (n = 43) revealed a diffuse, rather than regional, quantitative distribution of spermatogenesis. A microsurgical approach resulted in the minimal amount of tissue loss and minimal-to-no pain (compared with the original 45 cases already reported). By this means it is often possible to immediately locate the few tubules with spermatogenesis at microsurgery, under local anaesthesia. But even in cases where greater amounts of tissue must be removed in order to find spermatozoa, the microsurgical TESE procedure prevents secondary testicular damage by protecting blood supply and preventing pain and atrophy from increased testicular pressure. Thus, future attempts at TESE-ICSI need not be compromised. PMID- 11056120 TI - Reactivity of different LH and FSH standards and preparations in the world health organization matched reagents for enzyme-linked immunoassays of gonadotrophins. AB - The immunoreactivity of various LH and FSH calibration standards and recombinant preparations in the enzyme-linked immunoassay (EIA) systems for gonadotrophins developed for the Special Programme of Research in Human Reproduction of the World Health Organization (WHO) were compared. The preparations tested included three LH and two FSH pituitary standards (calibrated against LH 80/552 and 68/40 and FSH 78/549 respectively) provided with the EIA or radioimmunoassay WHO matched reagent kits, the pituitary preparation LER-907, and recombinant human LH (rhLH) and FSH (rhFSH). Simultaneous curve fitting of the EIA dose-response curves revealed no significant differences among the slopes generated by the WHO LH standards and LER-907; in contrast, no parallelism was found between the curves of rhLH and the pituitary-derived LH standards. No significant differences were found among the slopes of the curves elicited by the pituitary and recombinant FSH preparations. Each LH preparation exhibited a high degree of charge heterogeneity. Considerable variations in charge isoform distribution among the WHO LH standards, rhLH and LER-907 were also evident. In contrast, the FSH preparations were less heterogeneous and exhibited minor differences in charge distribution. Despite the existing differences in charge isoform distribution, all the pituitary-derived preparations as well as rhFSH seem appropriate for using as calibration standards in this particular EIA system. PMID- 11056121 TI - High doses of gonadotrophins combined with stop versus non-stop protocol of GnRH analogue administration in low responder IVF patients: a prospective, randomized, controlled trial. AB - Recent evidence suggests that early cessation of gonadotrophin releasing hormone analogue (GnRHa) administration may offer some benefit to low responder patients undergoing IVF. A prospective, randomized controlled trial was designed to evaluate whether early cessation of GnRHa in an ovarian stimulation regimen is more beneficial than just increasing the doses of gonadotrophins. Seventy low responder patients (less than three mature follicles in a previous cycle) with normal basal follicle stimulating hormone concentrations and a previous cancelled IVF cycle were randomly allocated into two protocols: (i) non-stop protocol: long GnRHa suppression with high doses of gonadotrophins, and (ii) stop protocol, in which GnRHa administration is stopped with the onset of menses, while gonadotrophin doses remained similar to the non-stop protocol. A significantly higher number of mature oocytes were obtained in the study group (stop protocol) compared to the control group (non-stop protocol) (8.7 +/- 0.9 versus 6.2 +/- 0.7, P: = 0. 027). The stop protocol reduced the number of ampoules of gonadotrophins required (56.6 +/- 2.7 versus 68.0 +/- 3.5, P: = 0. 013). Both protocols resulted in a similar cancellation rate (2.7 versus 5.8%) (with no cycles cancelled due to ovulation), pregnancy rate (14.3 versus 18.7%), and implantation rate (12.1 versus 8.8%). The early cessation of GnRHa combined with high doses of gonadotrophins permitted the retrieval of a significantly higher number of oocytes. PMID- 11056122 TI - Dynamic assays of inhibin B and oestradiol following buserelin acetate administration as predictors of ovarian response in IVF. AB - The study was designed to examine whether dynamic measurements of inhibin B and oestradiol following single administration of buserelin acetate were correlated with the ovarian response to stimulation in IVF. A total of 37 patients undergoing IVF treatment was studied when the long protocol was started in the early follicular phase. Blood samples were taken twice: on day 2 of the menstrual cycle, before the first s.c. administration of buserelin acetate 0.5 mg and on day 3, 24 h later. Inhibin B and oestradiol concentrations were compared with the ovarian response to stimulation. The ovarian response was defined in two ways: 'number of oocytes/total recombinant (r) follicle stimulating hormone (FSH) dose'; and 'square-root (number of follicles/total rFSH dose)'. The following measurements were highly correlated with the ovarian response to stimulation: increase in oestradiol (day 3-day 2 oestradiol concentration) [correlation coefficient (r) = 0.68, P: < 0.0001] and sum of inhibin B (day 2 + day 3 inhibin B concentrations) (r = 0.6, P: < 0.0001). Age and basal concentrations of FSH and inhibin B were inferior to the above measurements in terms of correlation with the ovarian response. In conclusion, dynamic measurements of inhibin B and oestradiol following single administration of buserelin acetate were highly correlated with the ovarian response to stimulation for IVF treatment. PMID- 11056123 TI - Clomiphene citrate increases insulin-like growth factor binding protein-1 and reduces insulin-like growth factor-I without correcting insulin resistance associated with polycystic ovarian syndrome. AB - The induction of ovulation by clomiphene could be the result of interaction of the drug at various levels: hypothalamus, pituitary and ovary. It was demonstrated that administration of clomiphene to women with polycystic ovarian syndrome (PCOS) is accompanied by a reduction in plasma concentrations of insulin like growth factor-I (IGF-I). IGF-I seems to have an overall negative effect on normal folliculogenesis and ovulation. The aim of the present study was to evaluate the effect of clomiphene on plasma concentrations of IGF-I and IGF binding protein (IGFBP)-1 and on insulin resistance associated with PCOS. Fifteen patients diagnosed with PCOS were recruited. Clinical diagnosis was based on chronic oligomenorrhoea or amenorrhoea and hyperandrogenaemia. Clomiphene citrate was administered at a dose of 100mg/day to all women from day 5 to day 9 of the spontaneous or medroxyprogesterone acetate (MAP)-induced menstrual cycle. Blood sampling and a 2 h oral glucose loading test (75 g) were performed the day before and after the course of clomiphene. Ovulation was confirmed in 13/15 PCOS patients. Plasma concentrations of IGF-I decreased by 31.5% (434 +/- 84 versus 297 +/- 71 ng/ml; P: < 0.05) after 5 days of clomiphene therapy, whereas plasma concentrations of IGFBP-1 increased by approximately 28.1% (26.3 +/- 4 versus 36.6 +/- 7 ng/ml; P: < 0.05). This gave a 56.5% reduction in the IGF-I:IGFBP-1 ratio (21.9 versus 9.53). No significant changes in basal plasma concentrations of fasting insulin or area under the insulin curve were observed in response to oral loading. The present results show that clomiphene does not cause changes in insulin resistance associated with PCOS but reduces plasma concentrations of IGF I and increases those of IGFBP-1, with a consequent marked reduction in the IGF I:IGFBP-1 ratio. PMID- 11056124 TI - The vascular character of ovarian follicular granulosa cells: phenotypic and functional evidence for an endothelial-like cell population. AB - Ovarian follicular granulosa cells express temporally and spatially distinct functions throughout the follicle cycle. During the entire cycle, granulosa cells exhibit an unusually broad range of activities including the secretion of steroid hormones, enzymes, growth factors and cytokines. To date, the identity(ies) of these cells (lineage/cell type) remains unknown. We demonstrate expression of the Tie, Tek, cKit, Flt-1, CD-31 and vWF proteins and the ability to rapidly internalize acetylated low density lipoprotein among mural and cumulus subpopulations of human and murine follicular granulosa cells. In addition, we provide evidence that human and murine granulosa cells can engage in tube-forming activity in vitro. To the best of our knowledge, the six phenotypic and two functional markers examined during this study, as a group, are associated only with endothelial or endothelial-like cells. In total, the findings suggest that some granulosa cells may have the potential to actively participate in the vascularization of the corpus luteum, by way of an inherent capacity which is likely to be a characteristic of their unique identity and lineage. This inherent capacity of granulosa cells to behave and respond, at least to some extent, like endothelial cells may be of possible importance in the aetiology of certain follicular pathologies. PMID- 11056125 TI - Evaluation of the effects of extremely low frequency electromagnetic fields on mammalian follicle development. AB - The aim of this study was to evaluate the effects of pulsed, extremely low frequency electromagnetic fields (ELF-EMF) on in-vitro mouse pre-antral follicle development. Pre-antral follicles were cultured for 5 days and exposed to ELF-EMF at the frequencies of 33 or 50 Hz. ELF-EMF application did not affect follicular growth over a 3 day culture period, but on day 5 the growth of 33 Hz-exposed follicles was significantly reduced when compared with controls, while the 50 Hz exposed follicles were not significantly affected. However, ELF-EMF severely impaired antrum formation at both frequencies, as 79 +/- 3% of control follicles developed antral cavities compared with 30 +/- 6% and 51.6 +/- 4% of 33 or 50 Hz exposed follicles respectively. The follicles with failed antrum formation showed lower oestradiol release and granulosa cell DNA synthesis, but these effects were not related to granulosa cell apoptosis. Furthermore, a high percentage of the in vitro grown oocytes obtained from exposed follicles had a reduced ability to resume meiotic maturation when compared with controls. These results suggest that ELF-EMF exposure might impair mammalian female reproductive potentiality by reducing the capacity of the follicles to reach a developmental stage that is an essential pre-requisite for reproductive success. PMID- 11056126 TI - Gestational surrogacy: a feasible option for patients with Rokitansky syndrome. AB - Rokitansky syndrome is a developmental defect characterized by agenesis of the uterus and vagina but normal gonads and secondary sexual characters. It is not commonly transmitted as a dominant genetic trait. Surrogacy, which is legally and ethically accepted in the UK and other countries, has made it possible for the patients with this syndrome to have their own genetic children. Six patients with Rokitansky syndrome underwent 11 ovarian stimulation cycles that resulted in 11 fresh and three frozen embryo transfer procedures into six prospective surrogate mothers. Both commissioning and surrogate couples were properly screened and counselled and their treatment was approved by the clinic internal review committee (ethics committee). The treatment cycles resulted in six clinical pregnancies (42.9% pregnancy rate per embryo transfer and 54.5% per oocyte retrieval) and three live births (21. 4% per embryo transfer, 27.3% per retrieval and 50% per patient). Gestational surrogacy is a viable treatment for patients with Rokitansky syndrome. Such patients should be well informed and supported to be able to have a family using their own genetic gametes. PMID- 11056127 TI - Plasma leptin concentrations are increased in women with premenstrual syndrome. AB - Leptin is a metabolic regulator of the hypothalamic- pituitary-gonadal axis, and plays an important role in human reproduction. Its neuro-endocrine effects are mediated by interactions with receptors in the hypothalamus, where emotional drive is also controlled. We postulated that circulating leptin concentrations are increased in premenstrual syndrome (PMS), and that this may be associated with the psychological symptoms of the disease. We obtained fasting venous samples from 32 women with PMS and 28 women with asymptomatic menstrual cycles, matched for age, body mass index and menstrual cycle length. Leptin concentrations were measured by radioimmunoassay. Leptin concentrations increased significantly during the luteal phases of the menstrual cycles of the control and PMS groups as compared with the follicular phase, having excluded the 11 women with PMS and six controls found to be anovulatory on the basis of mid-luteal plasma progesterone concentrations from the analysis. A greater increase was observed in women with PMS than the controls (P: = 0.00006 and 0.003 respectively). Although leptin concentrations in the follicular and luteal phases were higher in PMS than the controls, the difference was only statistically significant between the follicular phases (P: = 0.001). There was no clear relationship between leptin and oestradiol or progesterone in this study. These findings suggest that leptin may play a role in the pathophysiology of the disease, and requires further evaluation. PMID- 11056128 TI - Dose-finding study of triptorelin acetate for prevention of a premature LH surge in IVF: a prospective, randomized, double-blind, placebo-controlled study. AB - Gonadotrophin-releasing hormone agonists (GnRHa) are routinely used in IVF programmes to prevent an unwanted LH surge and consequent ovulation. Despite its widespread use in IVF, a convincing dose recommendation for GnRHa in IVF does not exist. In our opinion, the lowest possible dose of GnRHa should be used. Thus, we performed a prospective, randomized, double-blind, placebo-controlled study to determine the minimal daily dose of triptorelin acetate needed to suppress a premature LH surge during IVF treatment in a long protocol. A total of 240 women (60 in each group) was randomized to either placebo or to one of three doses of triptorelin, i.e. 15, 50 or 100 microg daily. Ovarian stimulation was performed with two or three ampoules of FSH daily. A premature LH surge occurred in 23% of placebo-treated patients, but in none of the triptorelin acetate-treated patients. There were significantly more oocytes and embryos in the 50 and 100 microg triptorelin groups. There was no dose relationship in rates of either implantation, pregnancy, ongoing pregnancy, live birth or baby take-home. In this study we showed that daily administration of 15 microg triptorelin is sufficient to prevent a premature LH surge, and that 50 microg is equivalent to 100 microg in terms of IVF results. PMID- 11056129 TI - Day 3 serum inhibin B and FSH and age as predictors of assisted reproduction treatment outcome. AB - Recent reports investigating the value of basal inhibin B determination as a predictor of ovarian reserve and assisted reproduction treatment have led to discordant results. This study was undertaken to further assess the relative power of day 3 inhibin B and follicle stimulating hormone (FSH) (defined before treatment) and the woman's age both as single and combined predictors of ovarian response and pregnancy in an in-vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) programme. A total of 120 women undergoing their first cycle of IVF or ICSI was included. Forty consecutive cycles cancelled because of poor follicular response were initially selected. As a control group, the nearest completed IVF/ICSI cycles before and after each cancelled cycle (i.e. the closest cycles in temporal relationship to the index cycle) were used. Mean age and basal FSH concentrations were significantly higher in the cancelled than in the control group (P: < 0.01 and P: < 0.001 respectively), whereas basal inhibin B was significantly higher in the latter (P: < 0.05). The association of basal FSH (with an accuracy or predictive value of ovarian response of 79%) with cancellation rate was significant, independent of, and stronger than the effects of age and inhibin B (P: < 0.05). Any two or all three of these variables studied did not improve the predictive value of FSH alone. Woman's age was the only variable independently associated with pregnancy rate. It is concluded that the stronger predictors of success in patients undergoing their first IVF/ICSI treatment cycle are age and basal FSH rather than inhibin B. Basal FSH concentration was a better predictor of cancellation rate than age, but age was a stronger predictor of pregnancy rate. PMID- 11056130 TI - Successful transfer of frozen-thawed embryos obtained immediately before radical surgery for stage IIIa serous borderline ovarian tumour: case report. AB - A stage IIIA borderline serous ovarian tumour was treated conservatively by laparoscopy to preserve the fertility of a 21 year old nulligravid woman. Six months later, recurrent lesions were resected. An 'urgent' IVF was performed to obtain frozen embryos. Oncological treatment was then completed by radical surgery with uterine conservation. Fifteen months later, two thawed embryos were successfully transferred and the patient delivered one baby. From this observation, the authors discuss an alternative to oocyte donation in cases of bilateral ovariectomy for stage IIIA borderline serous ovarian tumour. PMID- 11056131 TI - Effect of the Yuzpe regimen of emergency contraception on markers of endometrial receptivity. AB - This exploratory study was designed to determine whether treatment with the Yuzpe regimen of emergency contraception altered endometrial integrin expression or other markers of uterine receptivity. Nineteen parous women were followed for two menstrual cycles. In the second cycle, each participant took 100 mg ethinyl oestradiol and 1 mg norgestrel on the day of the urinary luteinizing hormone (LH) surge and repeated the dose 12 h later. In both cycles, endometrial biopsy, phlebotomy and vaginal sonogram were performed 8-10 days after the urinary LH surge. No significant difference was found between untreated and treated cycles in most measures of endometrial histology or in endometrial expression of beta3 integrin subunit, leukaemia inhibitory factor, glycodelin, or progesterone receptors assessed by immunohistochemical techniques. Five statistically significant changes were noted in treated cycles: a reduction in endometrial MUC 1 expression, an increase in endometrial oestrogen receptor, lower luteal phase serum oestrogen concentration, reduced endometrial thickness, and greater proportion of glandular supranuclear vacuoles. The relationship of these findings to the contraceptive action of the Yuzpe regimen is unclear. PMID- 11056132 TI - Beyond the clinical classification of azoospermia: opinion. AB - There is an ongoing debate regarding the appropriate classification of azoospermia. This manuscript reviews the rationale for the current classification of azoospermia and how to effect a change if there is a need to do so. The current classification of azoospermia into obstructive and non-obstructive is because azoospermia due to ejaculatory duct dysfunction and hypogonadotrophism are extremely rare. Though the use of clinical protocols (defective spermatogenesis, genital tract obstruction, ejaculatory duct dysfunction, hypogonadotrophism or pre-testicular, testicular and post-testicular) may be useful in selecting patients for appropriate treatment, no study has shown that they provide a better method of classification of azoospermia than the current approach. There is increasing evidence of a genetic basis of male infertility as well as the evidence that men's fertility potential may be classified genetically. Moreover, genetic disorders may be transmitted to the offspring and their presence in infertile couples may affect treatment outcome. It is therefore useful to explore a genetic classification of azoospermia. PMID- 11056133 TI - Intra- and inter-individual variability in human sperm concentration, motility and vitality assessment during a workshop involving ten laboratories. AB - The aim of the present study was to assess variability in the evaluation of human sperm concentration, motility and vitality. Technicians and biologists from 10 teams involved in multicentre studies on semen quality attended the same laboratory, each team using its own methods and equipment to analyse the same semen samples. Inter-individual variability was assessed from 17 fresh semen samples of varying quality. Intra-individual variability was assessed from pools of frozen samples for sperm concentration and motility and stained smears for vitality with three blind evaluations by sample and smear. The mean inter individual coefficients of variation were 22.9, 21.8 and 17.5% for sperm concentration, motility and vitality respectively. There was no statistical difference among participants for sperm concentration assessment, but significant differences for both motility and vitality (both P: < 0.05). The mean intra individual coefficients of variation were 15.8, 26.2 and 13.1% for sperm concentration, motility and vitality respectively, with marked differences between expert and novice participants: concentration 9.8% versus 28.0%; motility 22.8% versus 33.0%; and vitality 10.0% versus 19.3%. The present data confirm the need for external quality control schemes for diagnostic purposes, and indicate their utmost importance in multicentre studies on semen quality. PMID- 11056134 TI - Offspring sex ratio of subfertile men and men with abnormal sperm characteristics. AB - Previous work has suggested an association between male subfertility and a female biased offspring sex ratio. This study of the reproduction of men who had a semen analysis at the Sperm Analysis Laboratory in Copenhagen in the period 1963-1993 showed that the subfertile men had an offspring sex ratio as expected (51.0% boys versus 51.3%, P: = 0.56), and within the cohort, the offspring sex ratio had no material association with particular semen characteristics. Our results thus suggest that no important association exists between general male subfertility and a female-biased offspring sex ratio. PMID- 11056135 TI - Increased sperm motility after in-vitro culture of testicular biopsies from obstructive azoospermic patients results in better post-thaw recovery rate. AB - The objective of this study was to optimize the use of testicular biopsies in 14 patients with obstructive azoospermia. Testicular specimens were retrieved from six patients (group I) and cultured at 32 and 37 degrees C for up to 20 days; changes in percentage motile spermatozoa were compared. In four men of group I, one portion of the specimen was frozen at retrieval, and changes in post-thaw motility after 24 h of culture at 37 degrees C were recorded. In the other eight patients (group II), testicular specimens were frozen at retrieval and after 72 h culture at 37 degrees C. Pre and post-freezing motility and post-thaw recovery rate were compared. No significant differences were observed until day 8 in the improvement of motility between 32 and 37 degrees C in-vitro culture. Maximum motility was reached, under both conditions, between 48 h and 72 h. Post-thaw 24 h culture at 37 degrees C of specimens frozen at retrieval did not improve motility; however, 72 h pre-freezing culture significantly improved initial motility (P: < 0.01), post-thaw motility (P: < 0.01) and post-thaw recovery rate (P: < 0. 001). The higher recovery rate of samples frozen 3 days after retrieval allows more economical use of the tissue that is available. PMID- 11056136 TI - Thalassaemic men affected by erectile dysfunction treated with transurethral alprostadil: case report. AB - The aim of this controlled clinical study, performed in a specialized institutional unit for thalassaemic men, was to consider the possibility of restoring erection in beta-thalassaemic patients with erectile dysfunction by administering E(1) prostaglandins (alprostadil) transurethrally. Four patients affected by beta-thalassaemia, aged between 32 and 52 years, and having an erectile dysfunction were included in the study. Each patient was given 500 microg alprostadil in the distal urethra. Response was evaluated by the erection assessment scale. The main outcome measures were: (i) the clinical study; (ii) FSH, LH, total and free testosterone plasma concentrations; and (iii) basal and dynamic Doppler sonography of cavernous arteries. The treatment produced a response of 3-4 on the erection assessment scale. Average minimum response time was 20 min, while average maximum response time was about 60 min. There was no evidence of significant side effects. Our hypothesis is that the delayed reaction was due to organ damage induced by iron load, causing a reduction or absence of elasticity in the interstitial tissue of the corpora cavernosa. Thus, we believe that treatment with alprostadil can be considered an effective, non-invasive therapy for thalassaemic patients with erectile dysfunction. PMID- 11056137 TI - Transvaginal hydrolaparoscopy compared with laparoscopy for the evaluation of infertile women: a prospective comparative blind study. AB - Standard diagnostic laparoscopy is considered the gold standard to investigate tubo-peritoneal infertility. It requires general anaesthesia and full operative facilities. Due to the risk of complications, laparoscopy is frequently postponed to the final stage of infertility evaluation or even after treatment trials have failed. Transvaginal hydrolaparoscopy (THL) is based on vaginal access using a needle puncture technique and saline for distention. THL can be performed on an outpatient basis under local anaesthesia. However, little data exist concerning the accuracy of THL in comparison with laparoscopy. We conducted a prospective comparative blind trial to assess the feasibility and accuracy of THL compared with diagnostic laparoscopy in infertile women. Sixty women were assigned to undergo THL immediately prior to laparoscopy. Different operators evaluated the findings of the two procedures. In order to evaluate the accuracy of THL, findings in terms of tubal pathology, endometriosis and adhesions were analysed. The success rate of accessing the pouch of Douglas was 90.2%. Complication rate was 1.6%. THL diagnosis was correlated with that of laparoscopy in 92.3% of cases. In cases of abnormal findings by THL, there were no normal laparoscopies. Our pilot study suggests that THL is a safe and reproducible method. Retroverted uterus should be considered as a relative contraindication to THL. When a complete evaluation by THL is available, it is a highly accurate technique in comparison with the laparoscopy. PMID- 11056138 TI - Treatment of normal women with oestradiol plus progesterone prevents the decrease of leptin concentrations induced by ovariectomy. AB - To study the role of oestradiol and progesterone in the secretion of leptin, 21 normally ovulating women were recruited from those scheduled for ovariectomy plus hysterectomy performed in mid-follicular phase of the cycle. Seven of the women were used as controls and received no hormonal treatment post-operatively. Another seven women received oestradiol (oestradiol group) and the remaining seven women received oestradiol plus progesterone (oestradiol plus progesterone group). Serum leptin values showed a temporal but significant increase 24 h after the operation and were significantly correlated with the cortisol and progesterone values, which increased temporarily at 12 h. At that time a marked decline in oestradiol concentrations was seen. After the temporal increase, leptin values in the controls and the oestradiol group decreased significantly up to day 4 (P: < 0.05), while in the oestradiol plus progesterone group they increased (P: < 0.01) and were significantly higher than in the other two groups (P: < 0.05). Body mass index (BMI) was the most important variable accounting for the changes in leptin values post-operatively, but in the oestradiol plus progesterone group progesterone correlated significantly with leptin independently of BMI. These results suggest that progesterone and cortisol can stimulate leptin secretion in women regardless of oestradiol concentrations. PMID- 11056139 TI - Turner's syndrome with concomitant hypopituitarism: case report. AB - The unique case of a young woman with concomitant pituitary insufficiency and gonadal dysgenesis due to Turner's syndrome is described. At the age of 17 years, when first examined elsewhere, this patient was prepubertal and short and a diagnosis of growth hormone deficiency was made. One year later, while on growth hormone (GH) substitution, thyrotrophin deficiency and hypogonadotrophic hypogonadism were confirmed and thyroxine and sex steroid substitution therapy was initiated. Upon evaluation in our clinic, at the age of 30 years, the low final height achieved with the GH substitution therapy, a number of clinical characteristics and the absence of ovarian tissue on ultrasound led to examination of the patient's karyotype, which revealed concurrent gonadal dysgenesis due to Turner's syndrome. This case illustrates that the co-existence of primary and secondary hypogonadism may obscure and delay the diagnosis of Turner's syndrome, a diagnosis which alters the counselling of the patient from the reproductive perspective. PMID- 11056140 TI - Relationship between granular cytoplasm of oocytes and pregnancy outcome following intracytoplasmic sperm injection. AB - Couples undergoing intracytoplasmic sperm injection (ICSI) for male infertility using oocytes with centrally located granular cytoplasm (CLCG) were evaluated for fertilization, embryo development, implantation and pregnancy rate. CLCG is a rare morphological feature of the oocyte, that is diagnosed as a larger, dark, spongy granular area in the cytoplasm. Severity is based on both the diameter of granular area and the depth of the lesion. Twenty-seven couples with 39 cycles presenting CLCG in >50% of retrieved oocytes were evaluated. A total of 489 oocytes was retrieved, out of which 392 were at MII. CLCG was observed in 258 of the MII oocytes (65. 8%); 66.7% of these oocytes had slight and 33.3% had severe CLCG. The overall fertilization rate was 72.2% and no statistical significant difference was found between normal and CLCG oocytes and between the oocytes representing slight and severe CLCG. The development and quality of embryos was the same in normal and CLCG oocytes. In nine cycles, preimplantation genetic diagnosis was executed to evaluate a possible accompanying chromosomal abnormality. Out of 44 blastomeres biopsied, 23 had chromosomal abnormality (52. 3%). Eleven pregnancies were achieved in 39 cycles (28.2%), six pregnancies resulted in abortion (54.5%). The implantation rate was found to be 4.2%. Only five ongoing pregnancies were achieved in 39 cycles (12.8%). Couples with CLCG oocytes should be informed about poor on-going pregnancy rates even if fertilization, embryo quality and total pregnancy rates are normal. Furthermore, a high aneuploidy rate may be linked to a high abortion rate. PMID- 11056141 TI - The morphology of human pronuclear embryos is positively related to blastocyst development and implantation. AB - Human embryos are selected for transfer using morphology at the cleaving and blastocyst stages. Zygote morphology has been related to implantation and pregnancy. The aim of this study was to relate pronuclear morphology to blastocyst development. Zygotes were scored according to distribution and size of nucleoli within each nucleus. Zygotes displaying equality between the nuclei had 49.5% blastocyst formation and those with unequal sizes, numbers or distribution of nucleoli had 28% blastocyst formation. Cleaving embryos that were selected initially by zygote morphology and secondarily by morphology on day 3 had increased implantation (IR) and pregnancy rates (PR; 31 and 57%), compared with those selected by morphology alone (19 and 33% respectively; P: < 0.01). There was a significant difference between zygote-scored and non-scored cycles on day 3 (PR: 57 versus 33%; IR: 31 versus 19%) and on day 5 (PR: 73 versus 58%; IR; 52 versus 39%). Zygote scoring can maintain pregnancy rates for both day 3 and day 5 transfers, increase implantation rates and reduce the numbers of embryos required to achieve a pregnancy. PMID- 11056142 TI - Spontaneous blastomere fusion after freezing and thawing of early human embryos leads to polyploidy and chromosomal mosaicism. AB - The incidence of blastomere fusion after cryopreservation of early human embryos (day 2 and day 3) was investigated using the standard propanediol technique. The process of fusion was observed in all developmental stages (from 2 to 10 cells) and the frequency of this event was 4.6% in day 2 (41/889) and 1.5% in day 3 (10/646) embryos that survived the thawing (embryos with 50-100% intact cells). Fusion of two, and occasionally of several, blastomeres resulted in the formation of multinucleated hybrid cells, which clearly indicated that the ploidy of these newly created cells had been altered. This event, depending on the number of fused cells per embryo, transformed the embryos into either entirely polyploid embryos (complete fusion at 2- or 3-cell stage) or into mosaics being a mixture of polyploid and normal cells. Chromosomal preparations of embryos affected by blastomere fusion indicated the presence of tetraploid mitotic plates. Also, fluorescence in-situ hybridization (FISH) analysis using DNA probes targeting unique sequences on chromosomes 9, 15, 17 and 22 indicated the existence of tetraploid and diploid fluorescence signals in the interphase nuclei within mosaics. Therefore, observations on live and fixed embryos suggested that tetraploid (4n) or hexaploid (6n) and tetraploid-diploid or more complex aberrations of ploidy might be formed as a consequence of blastomere fusion. Furthermore, this demonstrates that freezing and thawing may induce numerical chromosomal changes in human embryos. PMID- 11056143 TI - Birth of rhesus monkey infant after transfer of embryos derived from in-vitro matured oocytes: short communication. AB - Although strategies for in-vitro maturation of oocytes from rodents and domestic species have been relatively successful, application of these techniques to primates has not met with similar success. Currently, evaluation of the developmental capacity of oocytes following fertilization is the only reliable means to assess cytoplasmic maturation. Although rhesus monkey blastocysts have previously been produced from in-vitro matured oocytes, full developmental competence has not been demonstrated by term development. Here we report the birth of the first non-human primate infant derived from in-vitro matured oocytes. PMID- 11056144 TI - Cystic fibrosis in infertility: screening before assisted reproduction: opinion. AB - Cystic fibrosis (CF) is the most common autosomal recessive disease in Caucasians. In 97-98% of men with CF, bilateral congenital absence of the vas deferens (CBAVD) blocks the transport of spermatozoa resulting in azoospermia. Abnormalities in sperm parameters have also been identified in males with CF. To date, over 800 disease-causing mutations of the CF transmembrane conductance regulator (CFTR) gene have been identified (also called ABCC7). Current legislation suggests that prior to intracytoplasmic sperm injection (ICSI) treatment, men with CBAVD or unexplained oligozoospermia should be considered for screening. If the male is negative with routine screening then the female partner is not screened. This is fundamentally wrong because if the female is screened and is found to be CF positive on routine testing, her partner would then need the fullest possible investigation of the CFTR gene. It is ideal to screen both partners in cases of oligozoospermia. However, if the resources are stretched, then only the female needs to be routinely screened because if she is negative, then the couple's residual risk of having a CF or CBAVD child will be reduced to 1:960. Only when the female is found to be a carrier does the male partner need routine screening followed by full testing for known mutations. PMID- 11056145 TI - Female sex preponderance for idiopathic familial premature ovarian failure suggests an X chromosome defect: opinion. AB - Premature ovarian failure (POF) is defined as ovarian failure occurring before the age of 40 years. A genetic aetiology is suggested by the occurrence of families with two or more affected females. We have characterised the pattern of inheritance of 41 cases of familial POF and compared them to published pedigrees. In eleven families a clear genetic association of POF could be identified. In the remaining 30 families the mechanism of inheritance was obscure. We found a female sex preponderance in the siblings of 30 families with idiopathic POF and in previously published series of idiopathic familial POF. In contrast, other known causes of POF, such as blepharophimosis ptosis epicanthus and inversus and autosomal recessive gonadal dysgenesis, had no altered sex ratio. One of our series of 30 pedigrees demonstrated transmission of POF susceptibility through fathers, which we believe is the first to be described in the literature. We present a group of five consanguineous families where we assume the mode of inheritance is autosomal recessive and where there was no female sex preponderance. Female sex preponderance for idiopathic familial POF suggests an X chromosome defect is inherited as a major cause of ovarian failure. PMID- 11056146 TI - A study on placental transfer of diclofenac in first trimester of human pregnancy. AB - Diclofenac is a commonly used non-steroidal anti-inflammatory drug in women of reproductive age. It has teratogenic effects in animals. The aim of this study was to investigate the placental transfer of diclofenac in the first trimester of human pregnancy. Thirty patients undergoing surgical termination of pregnancy between 8 and 12 weeks gestation were given two doses of diclofenac before the procedure. Corresponding samples of maternal serum, amniotic fluid, coelomic fluid and fetal tissue were analysed by high-performance liquid chromatography. Diclofenac was detectable in all fetal tissue samples, with a concentration similar to that found in maternal venous samples. However, diclofenac was detectable in only 56.7 and 23.3% of the coelomic and amniotic fluid samples respectively, and the highest concentration attained was 80 and 5% of the maternal concentration respectively. In summary, we confirmed that diclofenac crosses the human placenta readily during the first trimester. Further studies are required to investigate the potential teratogenic effect of diclofenac in human embryos. PMID- 11056147 TI - Risk factors for 14-21 week abortions: a case-control study in Europe. The Europop Group. AB - Data from a case-control survey in Europe, carried out between 1994 and 1997, were used to investigate the risk factors for spontaneous abortions at 14-21 weeks (late abortions), according to the vital status of the fetus before the onset of labour. Late abortions included 62 involving a fetus alive before the onset of labour, 216 late abortions of a fetus already dead, together with 4592 control pregnancies at term (>/=37 weeks) from seven countries. Histories of induced abortion, spontaneous abortion and preterm birth were more closely associated with late abortion of a live fetus than with late abortion of a dead fetus. Women aged >/=35 years and women living alone had a much higher risk of late abortions than women aged 20-24 years and married women, regardless of the vital status of the fetus before labour. These results provide evidence that obstetric history and socio-demographic factors are risk factors for late abortions but differences are observed according to the vital status of the fetus before labour. PMID- 11056148 TI - The risks associated with pregnancy in women aged 35 years or older. AB - The obstetric risks of adverse outcome during pregnancy in women aged > or =35 years were quantified using a retrospective analysis of data from 385 120 singleton pregnancies in the North West Thames Region, UK, between 1988 and 1997. A comparison of pregnancy outcome was made on the basis of maternal age at delivery: 18-34 years (n = 336 462), 35-40 years (n = 41 327) and women aged > 40 years (n = 7331). Women aged <18 years (n = 5246) were excluded from the study. Data are presented as percentages of 18-34 year old women, 35-40 year old and > 40 year old women, with adjusted odds ratios (OR) according to age group. Pregnant women aged 35-40 years were at increased risk of: gestational diabetes, OR = 2.63 [99% confidence interval (CI) 2.40-2.89]; placenta praevia = 1.93 (1.58 2.35); breech presentation = 1.37 (1.28-1.47); operative vaginal delivery = 1.5 (1.43-1.57); elective Caesarean section = 1.77 (1.68-1.87); emergency Caesarean section = 1.59 (1.52-1.67); postpartum haemorrhage = 1.14 (1.09-1.19); delivery before 32 weeks gestation = 1.41 (1.24-1.61); birthweight below the 5th centile = 1.28 (1.20-1. 36); and stillbirth = 1.41 (1.17-1.70). Women aged >40 years had higher OR for the same risks. Pregnant women aged >/=35 years are at increased risk of complications in pregnancy compared with younger women. PMID- 11056149 TI - Three-dimensional qualitative sonographic evaluation of fetal soft tissue. AB - Even though fetal growth restriction and macrosomia remain as major problems currently facing obstetricians, there is still no modality for the assessment of fetal soft tissue deposition and muscle mass in utero. A total of 52 fetuses from 29 to 41 weeks gestation were studied within 1 week before delivery using a transabdominal three-dimensional (3D) transducer (3.5 MHz). Their birth weights varied from 1016 to 4018 g, and their crown-heel length from 37 to 54 cm. The amount of subcutaneous tissue was estimated using the fetal nutrition score. The fetal nutrition score values were determined from a qualitative assessment of the amount of subcutaneous tissue present at three locations (face, ribs and buttocks) on the antenatal 3D ultrasonograms. Fetal nutritional status, using fetal nutrition score, was compared with that found by modified neonatal nutrition score and ponderal index respectively. There was a significant linear correlation between fetal nutrition score and modified neonatal nutrition score. Fetal or neonatal nutrition score were strongly correlated with birth weight and neonatal crown-heel length. However, no significant correlation was found between ponderal index, fetal nutrition score, or modified neonatal nutrition score. Ponderal index also was not correlated with birth weight and neonatal crown-heel length. Moreover, fetal nutrition score was correlated with Apgar score, but not with umbilical cord arterial blood pH. Therefore, doubt is cast on the usefulness of the ponderal index for measurement of neonatal soft tissue and muscle mass. Fetal nutrition score using 3D ultrasonography provides a novel means of evaluating the nutritional status of the fetus in utero, and should be useful for predicting the extremes in fetal growth (fetal growth restriction and macrosomia) at an earlier stage than hitherto achieved. PMID- 11056150 TI - Pregnancy outcome in a patient with chronic malnutrition: case report. AB - This case report describes the management of a chronically malnourished woman during her first and second pregnancies. The emphasis of the management is on the investigation of her dysphagia and subsequent bypassing of her colonic interposition by the formation of a percutaneous gastrostomy. The case highlights spontaneous conception with a body mass index of 14 and the safety of enteral feeding during pregnancy. PMID- 11056151 TI - Evidence may change with more trials: concepts to be kept in mind. PMID- 11056152 TI - HLA-DR4 and pre-eclampsia. PMID- 11056153 TI - Use of ultrasound guidance during embryo transfer. PMID- 11056154 TI - Ring opening is not rate-limiting in the GTP cyclohydrolase I reaction. AB - GTP cyclohydrolase I catalyzes a mechanistically complex ring expansion affording dihydroneopterin triphosphate and formate from GTP. Single turnover quenched flow experiments were performed with the recombinant enzyme from Escherichia coli. The consumption of GTP and the formation of 5-formylamino-6-ribosylamino-2-amino 4(3H)-pyrimidinone triphosphate, formate, and dihydroneopterin triphosphate were determined by high pressure liquid chromatography analysis. A kinetic model comprising three consecutive unimolecular steps was used for interpretations where the first intermediate, 5-formylamino-6-ribosylamino-2-amino-4(3H) pyrimidinone 5'-triphosphate, was formed in a reversible reaction. The rate constant k(1) for the reversible opening of the imidazole ring of GTP was 0.9 s( 1), the rate constant k(3) for the release of formate from 5-formylamino-6 ribosylamino-2-amino-4(3H)-pyrimidinone triphosphate was 2.0 s(-1), and the rate constant k(4) for the formation of dihydroneopterin triphosphate was 0.03 s(-1). Thus, the hydrolytic opening of the imidazole ring of GTP is rapid by comparison with the overall reaction. PMID- 11056155 TI - G(i)-mediated Cas tyrosine phosphorylation in vascular endothelial cells stimulated with sphingosine 1-phosphate: possible involvement in cell motility enhancement in cooperation with Rho-mediated pathways. AB - Since blood platelets release sphingosine 1-phosphate (Sph-1-P) upon activation, it is important to examine the effects of this bioactive lipid on vascular endothelial cell functions from the viewpoint of platelet-endothelial cell interactions. In the present study, we examined Sph-1-P-stimulated signaling pathways related to human umbilical vein endothelial cell (HUVEC) motility, with a special emphasis on the cytoskeletal docking protein Crk-associated substrate (Cas). Sph-1-P stimulated tyrosine phosphorylation of Cas, which was inhibited by the G(i) inactivator pertussis toxin but not by the Rho inactivator C3 exoenzyme or the Rho kinase inhibitor Y-27632. Fyn constitutively associated with and phosphorylated Cas, suggesting that Cas tyrosine phosphorylation may be catalyzed by Fyn. Furthermore, upon HUVEC stimulation with Sph-1-P, Crk, through its SH2 domain, interacted with tyrosine-phosphorylated Cas, and the Cas-Crk complex translocated to the cell periphery (membrane ruffles), through mediation of G(i) (Fyn) but not Rho. In contrast, tyrosine phosphorylation of focal adhesion kinase, and formation of stress fibers and focal adhesion were mediated by Rho but not G(i) (Fyn). Finally, Sph-1-P-enhanced HUVEC motility, assessed by a phagokinetic assay using gold sol-coated plates and a Boyden's chamber assay, was markedly inhibited not only by pertussis toxin (or the Fyn kinase inhibitor PP2) but also by C3 exoenzyme (or Y-27632). In HUVECs stimulated with Sph-1-P, these data suggest the following: (i) cytoskeletal signalings may be separable into G(i)-mediated signaling pathways (involving Cas) and Rho-mediated ones (involving FAK), and (ii) coordinated signalings from both pathways are required for Sph-1-P enhanced HUVEC motility. Since HUVECs reportedly express the Sph-1-P receptors EDG-1 (coupled with G(i)) and EDG-3 (coupled with G(13) and G(q)) and the EDG-3 antagonist suramin was found to block specifically Rho-mediated responses, it is likely that Cas-related responses following G(i) activation originate from EDG-1, whereas Rho-related responses originate from EDG-3. PMID- 11056156 TI - Positive torsional strain causes the formation of a four-way junction at replication forks. AB - The advance of a DNA replication fork requires an unwinding of the parental double helix. This in turn creates a positive superhelical stress, a (+)-DeltaLk, that must be relaxed by topoisomerases for replication to proceed. Surprisingly, partially replicated plasmids with a (+)-DeltaLk were not supercoiled nor were the replicated arms interwound in precatenanes. The electrophoretic mobility of these molecules indicated that they have no net writhe. Instead, the (+)-DeltaLk is absorbed by a regression of the replication fork. As the parental DNA strands re-anneal, the resultant displaced daughter strands base pair to each other to form a four-way junction at the replication fork, which is locally identical to a Holliday junction in recombination. We showed by restriction endonuclease digestion that the junction can form at either the terminus or the origin of replication and we visualized the structure with scanning force microscopy. We discuss possible physiological implications of the junction for stalled replication in vivo. PMID- 11056157 TI - ATP-stimulated release of interleukin (IL)-1beta and IL-18 requires priming by lipopolysaccharide and is independent of caspase-1 cleavage. AB - Interleukin (IL)-1beta and IL-18 are structurally similar proteins that require caspase-1 processing for activation. Both proteins are released from the cytosol by unknown pathway(s). To better characterize the release pathway(s) for IL-1beta and IL-18 we evaluated the role of lipopolysaccharide priming, of interleukin 1beta-converting enzyme (ICE) inhibition, of human purinergic receptor (P2X(7)) function, and of signaling pathways in human monocytes induced by ATP. Monocytes rapidly processed and released both IL-1beta and IL-18 after exogenous ATP. Despite its constitutive cytosolic presence, IL-18 required lipopolysaccharide priming for the ATP-induced release. Neither IL-1beta nor IL-18 release was prevented by ICE inhibition, and IL-18 release was not induced by ICE activation itself. Release of both cytokines was blocked completely by a P2X7 receptor antagonist, oxidized ATP, and partially by an antibody to P2X(7) receptor. In evaluating the signaling components involved in the ATP effect, we identified that the protein-tyrosine kinase inhibitor, AG126, produced a profound inhibition of both ICE activation as well as release of IL-1beta/IL-18. Taken together, these results suggest that, although synthesis of IL-1beta and IL-18 differ, ATP mediated release of both cytokines requires a priming step but not proteolytically functional caspase-1. PMID- 11056158 TI - Equilibrium unfolding of Bombyx mori glycyl-tRNA synthetase. AB - Unfolding of Bombyx mori glycyl-tRNA synthetase was examined by multiple spectroscopic techniques. Tryptophan fluorescence of wild type enzyme and an N terminally truncated form (N55) increased at low concentrations of urea or guanidine-HCl followed by a reduction in intensity at intermediate denaturant concentrations; a transition at higher denaturant was detected as decreased fluorescence intensity and a red-shifted emission. Solute quenching of fluorescence indicated that tryptophans become progressively solvent-exposed during unfolding. Wild type enzyme had stronger negative CD bands between 220 and 230 nm than the mutant, indicative of greater alpha-helical content. Urea or guanidine-HCl caused a reduction in ellipticity at 222 nm at low denaturant concentration with the wild type enzyme, a transition that is absent in the mutant; both enzymes exhibited a cooperative transition at higher denaturant concentrations. Both enzymes dissociate to monomers in 1.5 m urea. Unfolding of wild type enzyme is described by a multistate unfolding and a parallel two state unfolding; the two-state component is absent in the mutant. Changes in spectral properties associated with unfolding were largely reversible after dilution to low denaturant. Unfolding of glycyl-tRNA synthetase is complex with a native state, a native-like monomer, partially unfolded states, and the unfolded state. PMID- 11056159 TI - Role for de novo sphingoid base biosynthesis in the heat-induced transient cell cycle arrest of Saccharomyces cerevisiae. AB - The recent findings of sphingolipids as potential mediators of yeast heat stress responses led us to investigate their possible role in the heat-induced cell cycle arrest and subsequent recovery. The sphingolipid-deficient yeast strain 7R4 was found to lack the cell cycle arrest seen in the isogenic wild type. Furthermore, strain lcb1-100, which harbors a temperature-sensitive serine palmitoyltransferase, lacked increased de novo generated sphingoid bases upon heat stress. Importantly, this strain was found to lack the transient heat induced G0/G1 arrest. These results indicate a role for sphingolipids and specifically those generated in the de novo pathway in the cell cycle arrest response to heat. To determine the bioactive sphingolipid regulating this response, an analysis of key mutants in the sphingolipid biosynthetic and degradation pathways was performed. Strains deleted in sphingoid base kinases, sphingoid phosphate phosphatase, lyase, or dihydrosphingosine hydroxylase were found to display the cell cycle arrest. Also, the knockout of a fatty acyl elongation enzyme, which severely attenuates ceramide production, displayed the arrest. These experiments suggested that the active species for cell cycle arrest were the sphingoid bases. In further support of these findings, exogenous phytosphingosine (10 microM) was found to induce transient arrest. Stearylamine did not induce an arrest, demonstrating chemical specificity, and L-erythro- was not as potent as D-erythro-dihydrosphingosine showing stereospecificity. To investigate a possible arrest mechanism, we studied the hyperstable Cln3 (Cln3-1) strain LDW6A that has been previously shown to be resistant to heat stress induced cell cycle arrest. The strain containing Cln3-1 was found to be resistant to cell cycle arrest induced by exogenous phytosphingosine, indicating that Cln3 acts downstream of the sphingoid bases in this response. Interestingly, cell cycle recovery from the transient arrest was found to be dependent upon the sphingoid base kinases (LCB4, LCB5). Overall, this combination of genetic and pharmacologic results demonstrates a role for de novo sphingoid base biosynthesis by serine palmitoyltransferase in the transient G0/G1 arrest mediated through Cln3 via a novel mechanism. PMID- 11056160 TI - A novel alcohol oxidase/RNA-binding protein with affinity for mycovirus double stranded RNA from the filamentous fungus Helminthosporium (Cochliobolus) victoriae: molecular and functional characterization. AB - We have cloned and sequenced a novel alcohol oxidase (Hv-p68) from the filamentous fungus Helminthosporium (Cochliobolus) victoriae that copurifies with mycoviral double-stranded RNAs. Sequence analysis revealed that Hv-p68 belongs to the large family of FAD-dependent glucose methanol choline oxidoreductases and that it shares significant sequence identity (>67%) with the alcohol oxidases of the methylotrophic yeasts. Unlike the intronless alcohol oxidases from methylotrophic yeasts, a genomic fragment of the Hv-p68 gene was found to contain four introns. Hv-p68, purified from fungal extracts, showed only limited methanol oxidizing activity, and its expression was not induced in cultures supplemented with methanol as the sole carbon source. Northern hybridization analysis indicated that overexpression of Hv-p68 is associated with virus infection, because significantly higher Hv-p68 mRNA levels (10- to 20-fold) were detected in virus-infected isolates compared with virus-free ones. We confirmed by Northwestern blot analysis that Hv-p68 exhibits RNA binding activity and demonstrated that the RNA-binding domain is localized within the N-terminal region that contains a typical ADP-binding beta-alpha-beta fold motif. The Hv-p68 gene, or closely similar genes, was present in all species of the genus Cochliobolus but absent in the filamentous fungus, Penicillium chrysogenum, as well as in two nonmethylotrophic yeasts examined. This study represents the first reported case that a member of the FAD-dependent glucose methanol choline oxidoreductase family, Hv-p68, may function as an RNA-binding protein. PMID- 11056161 TI - Single turnover chemistry and regulation of O2 activation by the oxygenase component of naphthalene 1,2-dioxygenase. AB - Naphthalene 1,2-dioxygenase (NDOS) is a three-component enzyme that catalyzes cis (1R,2S)-dihydroxy-1,2-dihydronaphthalene formation from naphthalene, O2, and NADH. We have determined the conditions for a single turnover of NDOS for the first time and studied the regulation of catalysis. As isolated, the alpha3beta3 oxygenase component (NDO) has up to three catalytic pairs of metal centers (one mononuclear Fe2+ and one diferric Rieske iron-sulfur cluster). This form of NDO is unreactive with O2. However, upon reduction of the Rieske cluster and exposure to naphthalene and O2, approximately 0.85 cis-diol product per occupied mononuclear iron site rapidly forms. Substrate binding is required for oxygen reactivity. Stopped-flow and chemical quench analyses indicate that the rate constant of the single turnover product-forming reaction significantly exceeds the NDOS turnover number. UV-visible and electron paramagnetic resonance spectroscopies show that during catalysis, one mononuclear iron and one Rieske cluster are oxidized per product formed, satisfying the two-electron reaction stoichiometry. The addition of oxidized or reduced NDOS ferredoxin component (NDF) increases both the product yield and rate of oxidation of formerly unreactive Rieske clusters. The results show that NDO alone catalyzes dioxygenase chemistry, whereas NDF appears to serve only an electron transport role, in this case redistributing electrons to competent active sites. PMID- 11056162 TI - Solution structure of the N-terminal domain of the human TFIIH MAT1 subunit: new insights into the RING finger family. AB - The human MAT1 protein belongs to the cyclin-dependent kinase-activating kinase complex, which is functionally associated to the transcription/DNA repair factor TFIIH. The N-terminal region of MAT1 consists of a C3HC4 RING finger, which contributes to optimal TFIIH transcriptional activities. We report here the solution structure of the human MAT1 RING finger domain (Met(1)-Asp(65)) as determined by (1)H NMR spectroscopy. The MAT1 RING finger domain presents the expected betaalphabetabeta topology with two interleaved zinc-binding sites conserved among the RING family. However, the presence of an additional helical segment in the N-terminal part of the domain and a conserved hydrophobic central beta strand are the defining features of this new structure and more generally of the MAT1 RING finger subfamily. Comparison of electrostatic surfaces of RING finger structures shows that the RING finger domain of MAT1 presents a remarkable positively charged surface. The functional implications of these MAT1 RING finger features are discussed. PMID- 11056163 TI - Polymorphic deletion of three intracellular acidic residues of the alpha 2B adrenergic receptor decreases G protein-coupled receptor kinase-mediated phosphorylation and desensitization. AB - A polymorphic variant of the human alpha(2B)-adrenergic receptor (alpha(2B)AR), which consists of a deletion of three glutamic acids (residues 301-303) in the third intracellular loop was found to be common in Caucasians (31%) and to a lesser extent in African-Americans (12%). The consequences of this deletion were assessed by expressing wild-type and the Del301-303 receptors in Chinese hamster ovary and COS cells. Ligand binding was not affected, although a small decrease in coupling efficiency to the inhibition of adenylyl cyclase was observed with the mutant. The deletion occurs within a stretch of acidic residues that is thought to establish the milieu for agonist-promoted phosphorylation and desensitization of the receptor by G protein-coupled receptor kinases (GRKs). Agonist-promoted phosphorylation studies carried out in cells coexpressing the alpha(2B)ARs and GRK2 revealed that the Del301-303 receptor displayed approximately 56% of wild-type phosphorylation. Furthermore, the depressed phosphorylation imposed by the deletion was found to result in a complete loss of short term agonist-promoted receptor desensitization. Thus the major phenotype of the Del301-303 alpha(2B)AR is one of impaired phosphorylation and desensitization by GRKs, and thus the polymorphisms renders the receptor incapable of modulation by this key mechanism of dynamic regulation. PMID- 11056164 TI - Down-regulation of high mobility group-I(Y) protein contributes to the inhibition of nitric-oxide synthase 2 by transforming growth factor-beta1. AB - The inducible isoform of nitric-oxide synthase (NOS2) catalyzes the production of nitric oxide (NO), which participates in the pathophysiology of systemic inflammatory diseases such as sepsis. NOS2 is transcriptionally up-regulated by endotoxin and inflammatory cytokines, and down-regulated by transforming growth factor (TGF)-beta1. Recently we have shown that high mobility group (HMG)-I(Y) protein, an architectural transcription factor, contributes to NOS2 gene transactivation by inflammatory mediators. The aim of the present study was to determine whether regulation of HMG-I(Y) by TGF-beta1 contributes to the TGF beta1-mediated suppression of NOS2. By Northern blot analysis, we show that TGF beta1 decreased cytokine-induced HMG-I(Y) mRNA levels in vascular smooth muscle cells and macrophages in vitro and in vivo. Western analysis confirmed the down regulation of HMG-I(Y) protein by TGF-beta1. To determine whether the down regulation of HMG-I(Y) contributed to a decrease in NOS2 gene transactivation by TGF-beta1, we performed cotransfection experiments. Overexpression of HMG-I(Y) was able to restore cytokine inducibility of the NOS2 promoter that was suppressed by TGF-beta1. The effect of TGF-beta1 on NOS2 gene transactivation was not related to a decrease in binding of HMG-I(Y) to the promoter of the NOS2 gene, but due to a decrease in endogenous HMG-I(Y) protein. These data provide the first evidence that cytokine-induced HMG-I(Y) can be down-regulated by TGF beta1. This down-regulation of HMG-I(Y) contributes to the TGF-beta1-mediated decrease in NOS2 gene transactivation by proinflammatory stimuli. PMID- 11056165 TI - Multiple Yap1p-binding sites mediate induction of the yeast major facilitator FLR1 gene in response to drugs, oxidants, and alkylating agents. AB - The bZip transcription factor Yap1p plays an important role in oxidative stress response and multidrug resistance in Saccharomyces cerevisiae. We have previously demonstrated that the FLR1 gene, encoding a multidrug transporter of the major facilitator superfamily, is a transcriptional target of Yap1p. The FLR1 promoter contains three potential Yap1p response elements (YREs) at positions -148 (YRE1), -167 (YRE2), and -364 (YRE3). To address the function of these YREs, the three sites have been individually mutated and tested in transactivation assays. Our results show that (i) each of the three YREs is functional and important for the optimal transactivation of FLR1 by Yap1p and that (ii) the three YREs are not functionally equivalent, mutation of YRE3 being the most deleterious, followed by YRE2 and YRE1. Simultaneous mutation of the three YREs abolished transactivation of the promoter by Yap1p, demonstrating that the three sites are essential for the regulation of FLR1 by Yap1p. Gel retardation assays confirmed that Yap1p differentially binds to the three YREs (YRE3 > YRE2 > YRE1). We show that the transcription of FLR1 is induced upon cell treatment with the oxidizing agents diamide, diethylmaleate, hydrogen peroxide, and tert-butyl hydroperoxide, the antimitotic drug benomyl, and the alkylating agent methylmethane sulfonate and that this induction is mediated by Yap1p through the three YREs. Finally, we show that FLR1 overexpression confers resistance to diamide, diethylmaleate, and menadione but hypersensitivity to H(2)O(2), demonstrating that the Flr1p transporter participates in Yap1p-mediated oxidative stress response in S. cerevisiae. PMID- 11056166 TI - Identification of hypoxia-inducible factor 1 ancillary sequence and its function in vascular endothelial growth factor gene induction by hypoxia and nitric oxide. AB - Transcription of hypoxia-inducible genes is regulated by hypoxia response elements (HREs) located in either the promoter or enhancer regions. Analysis of these elements reveals the presence of one or more binding sites for hypoxia inducible factor 1 (HIF-1). Hypoxia-inducible genes include vascular endothelial growth factor (VEGF), erythropoietin, and glycolytic enzyme genes. Site-directed mutational analysis of the VEGF gene promoter revealed that an HIF-1 binding site (HBS) and its downstream HIF-1 ancillary sequence (HAS) within the HRE are required as cis-elements for the transcriptional activation of VEGF by either hypoxia or nitric oxide (NO). The core sequences of the HBS and the HAS were determined as TACGTG and CAGGT, respectively. These elements form an imperfect inverted repeat, and the spacing between these motifs is crucial for activity of the promoter. Gel shift assays demonstrate that as yet unknown protein complexes constitutively bind to the HAS regardless of the presence of these stimuli in several cell lines, in contrast with hypoxia- or NO-induced activation of HIF-1 binding to the HBS. A common structure of the HRE, which consists of the HBS and the HAS, is seen among several hypoxia-inducible genes, suggesting the presence of a novel mechanism mediated by the HAS for the regulation of these genes. PMID- 11056167 TI - Identification of functional domains and dominant negative mutations in vertebrate telomerase RNA using an in vivo reconstitution system. AB - The telomerase holoenzyme consists of two essential components, a reverse transcriptase, TERT (telomerase reverse transcriptase), and an RNA molecule, TR (telomerase RNA, also known as TERC), that contains the template for the synthesis of new telomeric repeats. Telomerase RNA has been isolated from 32 different vertebrates, and a common secondary structure has been proposed (Chen, J.-L., Blasco, M. A., and Greider, C. W. (2000) Cell 100, 503-514). We have generated 25 mutants in the four conserved structural domains of the mouse telomerase RNA molecule, mTR, and assayed their ability to reconstitute telomerase activity in mTR(-/-) cells in vivo. We found that the pseudoknot and the CR4/CR5 domains are required for telomerase activity but are not essential for mTR stability in the cell, whereas mutations in the BoxH/ACA and the CR7 domains affect mTR accumulation in the cell. We have also identified mTR mutants that are able to inhibit wild type telomerase in vivo. PMID- 11056168 TI - Two forms of UvrC protein with different double-stranded DNA binding affinities. AB - Using phosphocellulose followed by single-stranded DNA-cellulose chromatography for purification of UvrC proteins from overproducing cells, we found that UvrC elutes at two peaks: 0.4 m KCl (UvrCI) and 0.6 m KCl (UvrCII). Both forms of UvrC have a major peptide band (>95%) of the same molecular weight and identical N terminal amino acid sequences, which are consistent with the initiation codon being at the unusual GTG site. Both forms of UvrC are active in incising UV irradiated, supercoiled phiX-174 replicative form I DNA in the presence of UvrA and UvrB proteins; however, the specific activity of UvrCII is one-fourth that of UvrCI. The molecular weight of UvrCII is four times that of UvrCI on the basis of results of size exclusion chromatography and glutaraldehyde cross-linking reactions, indicating that UvrCII is a tetramer of UvrCI. Functionally, these two forms of UvrC proteins can be distinguished under reaction conditions in which the protein/nucleotide molar ratio is >0.06 by using UV-irradiated, (32)P-labeled DNA fragments as substrates; under these conditions UvrCII is inactive in incision, but UvrCI remains active. The activity of UvrCII in incising UV irradiated, (32)P- labeled DNA fragments can be restored by adding unirradiated competitive DNA, and the increased level of incision corresponds to a decreased level of UvrCII binding to the substrate DNA. The sites of incision at the 5' and 3' sides of a UV-induced pyrimidine dimer are the same for UvrCI and UvrCII. Nitrocellulose filter binding and gel retardation assays show that UvrCII binds to both UV-irradiated and unirradiated double-stranded DNA with the same affinity (K(a), 9 x 10(8)/m) and in a concentration-dependent manner, whereas UvrCI does not. These two forms of UvrC were also produced by the endogenous uvrC operon. We propose that UvrCII-DNA binding may interfere with Uvr(A)(2)B-DNA damage complex formation. However, because of its low copy number and low binding affinity to DNA, UvrCII may not interfere with Uvr(A)(2)B-DNA damage complex formation in vivo, but instead through double-stranded DNA binding UvrCII may become concentrated at genomic areas and therefore may facilitate nucleotide excision repair. PMID- 11056169 TI - Human p8 is a HMG-I/Y-like protein with DNA binding activity enhanced by phosphorylation. AB - We have studied the biochemical features, the conformational preferences in solution, and the DNA binding properties of human p8 (hp8), a nucleoprotein whose expression is affected during acute pancreatitis. Biochemical studies show that hp8 has properties of the high mobility group proteins, HMG-I/Y. Structural studies have been carried out by using circular dichroism (near- and far ultraviolet), Fourier transform infrared, and NMR spectroscopies. All the biophysical probes indicate that hp8 is monomeric (up to 1 mm concentration) and partially unfolded in solution. The protein seems to bind DNA weakly, as shown by electrophoretic gel shift studies. On the other hand, hp8 is a substrate for protein kinase A (PKA). The phosphorylated hp8 (PKAhp8) has a higher content of secondary structure than the nonphosphorylated protein, as concluded by Fourier transform infrared studies. PKAhp8 binds DNA strongly, as shown by the changes in circular dichroism spectra, and gel shift analysis. Thus, although there is not a high sequence homology with HMG-I/Y proteins, hp8 can be considered as a HMG-I/Y like protein. PMID- 11056170 TI - Two site contact of elongating transcripts to phage T7 RNA polymerase at C terminal regions. AB - A series of active elongation complexes of the phage T7 RNA polymerase were obtained through stepwise walking of the polymerase along an immobilized DNA template. Transcripts were radiolabeled at the 16th to 18th residues, and a photocross-linkable 4-thio-UMP was separately incorporated at the 22nd, 24th, 32nd, and 38th residues. Such complexes (up to 51 nucleotides) produced by the incorporation of one nucleotide at a time were isolated and individually subjected to long wave UV cross-linking. Only when the cross-linker was positioned at the 3'-end (-1) of the elongating RNA and 8 nucleotides upstream ( 9), was the RNA substantially cross-linked to the polymerase, regardless of how far it was from the 5'-end of the transcripts. Linkage of the 3'-end residue was mapped to the Thr(636)-Met(666) region, which contains nucleotide-binding sites. The -9 residue was cross-linked to the Ala(724)-Met(750) region rather than to the N-terminal region. These two contacts were maintained throughout the elongation complexes and reveal a route of nascent RNA through the T7 RNA polymerase in elongation complexes. PMID- 11056171 TI - Recruitment of the yeast Tup1p-Ssn6p repressor is associated with localized decreases in histone acetylation. AB - Posttranslational acetylation of histones is an important element of transcriptional regulation. The yeast Tup1p repressor is one of only a few non enzyme proteins known to interact directly with the amino-terminal tail domains of histones H3 and H4 that are subject to acetylation. We demonstrated previously that Tup1p interacts poorly with more highly acetylated isoforms of these histones in vitro. Here we show that two separate classes of promoters repressed by Tup1p are associated with underacetylated histones in vivo. This decreased histone acetylation is dependent upon Tup1p and its partner Ssn6p and is localized to sequences near the point of Tup1p-Ssn6p recruitment. Increased acetylation of histones H3 and H4 is observed upon activation of these genes, but this increase is not dependent on transcription per se. Direct recruitment of Tup1p-Ssn6p complexes via fusion of Tup1p to the lexA DNA binding domain is sufficient to confer repression and induce decreased acetylation of H3 and H4 at a target promoter. Taken together, our results suggest that stable decreases in histone acetylation levels are directed and/or maintained by the Tup1p-Ssn6p repressor complex. PMID- 11056172 TI - Electron transfer and binding of the c-type cytochrome TorC to the trimethylamine N-oxide reductase in Escherichia coli. AB - Reduction of trimethylamine N-oxide (E'(0(TMAO/TMA)) = +130 mV) in Escherichia coli is carried out by the Tor system, an electron transfer chain encoded by the torCAD operon and made up of the periplasmic terminal reductase TorA and the membrane-anchored pentahemic c-type cytochrome TorC. Although the role of TorA in the reduction of trimethylamine N-oxide (TMAO) has been clearly established, no direct evidence for TorC involvement has been presented. TorC belongs to the NirT/NapC c-type cytochrome family based on homologies of its N-terminal tetrahemic domain (TorC(N)) to the cytochromes of this family, but TorC contains a C-terminal extension (TorC(C)) with an additional heme-binding site. In this study, we show that both domains are required for the anaerobic bacterial growth with TMAO. The intact TorC protein and its two domains, TorC(N) and TorC(C), were produced independently and purified for a biochemical characterization. The reduced form of TorC exhibited visible absorption maxima at 552, 523, and 417 nm. Mediated redox potentiometry of the heme centers of the purified components identified two negative midpoint potentials (-177 and -98 mV) localized in the tetrahemic TorC(N) and one positive midpoint potential (+120 mV) in the monohemic TorC(C). In agreement with these values, the in vitro reconstitution of electron transfer between TorC, TorC(N), or TorC(C) and TorA showed that only TorC and TorC(C) were capable of electron transfer to TorA. Surprisingly, interaction studies revealed that only TorC and TorC(N) strongly bind TorA. Therefore, TorC(C) directly transfers electrons to TorA, whereas TorC(N), which probably receives electrons from the menaquinone pool, is involved in both the electron transfer to TorC(C) and the binding to TorA. PMID- 11056173 TI - Specific RNA binding by a single C2H2 zinc finger. AB - Zinc finger proteins with high affinity for human immunodeficiency virus Rev responsive element stem loop IIB (RRE-IIB) were previously isolated from a phage display zinc finger library. Zinc fingers from one of these proteins, RR1, were expressed individually and assayed for RRE-IIB affinity. The C-terminal zinc finger retained much of the binding affinity of the two-finger parent and was disrupted by mutations predicted to narrow the RRE-IIB major groove and which disrupt Rev binding. In contrast, the N-terminal zinc finger has a calculated affinity at least 1000-fold lower. Despite the high affinity and specificity of RR1 for RRE-IIB, binding affinity for a 234-nucleotide human immunodeficiency virus Rev responsive element (RRE234) was significantly lower. Therefore, zinc finger proteins that bind specifically to RRE234 were constructed using an in vitro selection and recombination approach. These zinc fingers bound RRE234 with subnanomolar dissociation constants and bound the isolated RRE-IIB stem loop with an affinity 2 orders of magnitude lower but similar to the affinity of an arginine-rich peptide derived from Rev. These data show that single C2H2 zinc fingers can bind RNA specifically and suggest that their binding to stem loop IIB is similar to that of Rev peptide. However, binding to RRE234 is either different from stem loop IIB binding or the tertiary structure of stem loop IIB is changed within the Rev responsive element. PMID- 11056174 TI - Contributions of Torpedo nicotinic acetylcholine receptor gamma Trp-55 and delta Trp-57 to agonist and competitive antagonist function. AB - Results of affinity-labeling studies and mutational analyses provide evidence that the agonist binding sites of the nicotinic acetylcholine receptor (nAChR) are located at the alpha-gamma and alpha-delta subunit interfaces. For Torpedo nAChR, photoaffinity-labeling studies with the competitive antagonist d [(3)H]tubocurarine (dTC) identified two tryptophans, gammaTrp-55 and deltaTrp-57, as the primary sites of photolabeling in the non-alpha subunits. To characterize the importance of gammaTrp-55 and deltaTrp-57 to the interactions of agonists and antagonists, Torpedo nAChRs were expressed in Xenopus oocytes, and equilibrium binding assays and electrophysiological recordings were used to examine the functional consequences when either or both tryptophans were mutated to leucine. Neither substitution altered the equilibrium binding of dTC. However, the deltaW57L and gammaW55L mutations decreased acetylcholine (ACh) binding affinity by 20- and 7,000-fold respectively. For the wild-type, gammaW55L, and deltaW57L nAChRs, the concentration dependence of channel activation was characterized by Hill coefficients of 1.8, 1.1, and 1.7. For the gammaW55L mutant, dTC binding at the alpha-gamma site acts not as a competitive antagonist but as a coactivator or partial agonist. These results establish that interactions with gamma Trp-55 of the Torpedo nAChR play a crucial role in agonist binding and in the agonist induced conformational changes that lead to channel opening. PMID- 11056175 TI - Two classes of androgen receptor elements mediate cooperativity through allosteric interactions. AB - Genes uniquely regulated by the androgen receptor (AR) typically contain multiple androgen response elements (AREs) that in isolation are of low DNA binding affinity and transcriptional activity. However, specific combinations of AREs in their native promoter context result in highly cooperative DNA binding by AR and high levels of transcriptional activation. We demonstrate that the natural androgen-regulated promoters of prostate specific antigen and probasin contain two classes of AREs dictated by their primary nucleotide sequence that function to mediate cooperativity. Class I AR-binding sites display conventional guanine contacts. Class II AR-binding sites have distinctive atypical sequence features and, upon binding to AR, the DNA structure is dramatically altered through allosteric interactions with the receptor. Class II sites stabilize AR binding to adjacent class I sites and result in synergistic transcriptional activity and increased hormone sensitivity. We have determined that the specific nucleotide variation within the AR binding sites dictate differential functions to the receptor. We have identified the role of individual nucleotides within class II sites and predicted consensus sequences for class I and II sites. Our data suggest that this may be a universal mechanism by which AR achieved unique regulation of target genes through complex allosteric interactions dictated by primary binding sequences. PMID- 11056176 TI - A map of protein-protein contacts within the small nuclear RNA-activating protein complex SNAPc. AB - The nucleation of RNA polymerases I-III transcription complexes is usually directed by distinct multisubunit factors. In the case of the human RNA polymerase II and III small nuclear RNA (snRNA) genes, whose core promoters consist of a proximal sequence element (PSE) and a PSE combined with a TATA box, respectively, the same multisubunit complex is involved in the establishment of RNA polymerase II and III initiation complexes. This factor, the snRNA-activating protein complex or SNAP(c), binds to the PSE of both types of promoters and contains five types of subunits, SNAP190, SNAP50, SNAP45, SNAP43, and SNAP19. SNAP(c) binds cooperatively with both Oct-1, an activator of snRNA promoters, and in the RNA polymerase III snRNA promoters, with TATA-binding protein, which binds to the TATA box located downstream of the PSE. Here we have defined subunit domains required for SNAP(c) subunit-subunit association, and we show that complexes containing little more than the domains mapped here as required for subunit-subunit contacts bind specifically to the PSE. These data provide a detailed map of the subunit-subunit interactions within a multifunctional basal transcription complex. PMID- 11056177 TI - Apicomplexan parasites possess distinct nuclear-encoded, but apicoplast localized, plant-type ferredoxin-NADP+ reductase and ferredoxin. AB - In searching for nuclear-encoded, apicoplast-localized proteins we have cloned ferredoxin-NADP(+) reductase from Toxoplasma gondii and a [2Fe-2S] ferredoxin from Plasmodium falciparum. This chloroplast-localized redox system has been extensively studied in photosynthetic organisms and is responsible for the electron transfer from photosystem I to NADP+. Besides this light-dependent reaction in nonphotosynthetic plastids (e.g. from roots), electrons can also flow in the reverse direction, from NADPH to ferredoxin, which then serves as an important reductant for various plastid-localized enzymes. These plastids possess related, but distinct, ferredoxin-NADP+ reductase and ferredoxin isoforms for this purpose. We provide phylogenetic evidence that the T. gondii reductase is similar to such nonphotosynthetic isoforms. Both the P. falciparum [2Fe-2S] ferredoxin and the T. gondii ferredoxin-NADP+ reductase possess an N-terminal bipartite transit peptide domain typical for apicoplast-localized proteins. The recombinant proteins were obtained in active form, and antibodies raised against the reductase recognized two bands on Western blots of T. gondii tachyzoite lysates, indicative of the unprocessed and native form, respectively. We propose that the role of this redox system is to provide reduced ferredoxin, which might then be used for fatty acid desaturation or other biosynthetic processes yet to be defined. Thus, the interaction of these two proteins offers an attractive target for drug intervention. PMID- 11056178 TI - Heme oxygenase (HO)-1 expressing macrophages/microglial cells accumulate during oligodendroglioma progression. AB - Heme oxygenase (HO-1, HSP32) catalyzes the oxidation of heme to biliverdin and carbon monoxide, a putative neurotransmitter. In the brain, HO-1 expression has been associated with neuroprotection during oxidative stress and hypoxia. However, consecutive downstream mediation is involved in neoangiogenesis and consequent neoplastic outgrowth. We have analyzed HO-1 expression in 69 oligodendroglioma tissue samples, in rat intracranially transplanted C6 gliomas, and neuropathologically unaltered control brains by immunohistochemistry. Double labeling experiments confirmed the nature of HO-1 expressing cells. Reverse transcription-polymerase chain reaction was used to demonstrate HO-1 gene expression. HO-1 immunoreactivity was predominantly observed in macrophages/microglial cells. The number of HO-1 expressing macrophages/microglial cells was significantly lower in primary oligodendrogliomas than in their matched relapses (P<0.0001) and lower in primary anaplastic oligodendrogliomas than in their relapses (P=0.0006). Prominent accumulation of HO-1 expressing macrophages/microglial cells was observed in perinecrotic areas of both experimental rat and human glioblastoma relapses. HO-1 expressing neurons, macrophages/microglial cells and astrocytes were scattered in areas of infiltrative tumor growth. Surprisingly, HO-1 mRNA was detected in only one glioblastoma multiforme relapse. We conclude from these data that HO-1 expressing macrophages/microglial cells accumulate during oligodendroglioma progression in areas of focal necrosis. However, overall biological function of this phenomenon remains to be determined. PMID- 11056179 TI - Protractive effects of chronic treatment with an acutely sub-toxic regimen of diisopropylflurophosphate on the expression of cholinergic receptor densities in rats. AB - Individuals chronically exposed to low levels of organophosphate insecticides may present with subtle impairments in cognition. In addition, low level diisopropylflurophosphate (DFP) exposure (0.25 mg/kg per day for 2 weeks) in rats resulted in protracted working memory impairment [29]. The current studies attempt to show a temporal relationship between the DFP-induced impairment in performance of a spatial memory task and the protracted decrease in the expression of cholinergic receptors and acetylcholinesterase in specific brain regions. Cholinergic receptors labeled with the ligands [(3)H]epibatidine and [(3)H]AFDX-384 were affected to a much greater extent and for a longer period of time than were both acetylcholinesterase activities and cholinergic receptors labeled with [(3)H]QNB. Pre-testing administration of nicotine was shown to completely reverse this DFP-induced impairment in memory-related task performance. Additionally, prophylaxis with pyridostigmine bromide (PB) caused DFP-treated animals to exhibit near normal levels of memory-related task performance. These results are consistent with the development of a protracted phase of learning impairment to sub-acute DFP exposure, which may involve the loss of hippocampal nicotinic receptors, and may be prevented or reversed by PB or nicotine, respectively. PMID- 11056180 TI - Plasminogen activation in experimental permanent focal cerebral ischemia. AB - BACKGROUND: Previous experimental work using in situ zymography has shown very early increased plasminogen activation in ischemic regions after 3 h of ischemia with and without reperfusion. The objective of the present study was to evaluate the time course and extent of plasminogen activation in long-term permanent focal cerebral ischemia. MATERIAL AND METHODS: The middle cerebral artery in male Fisher rats was irreversibly occluded by electrocoagulation. Duration of ischemia was 48, 72, and 168 h. Occlusion was controlled in vivo by MRI at day 2. Plasminogen activation was detected by in situ zymography of 10 microm cryosections with an overlay containing plasminogen and the plasmin substrate caseine. Areas of plasminogen activation were compared to structural lesions (immunohistochemical loss of microtubule-associated protein 2; MAP 2). RESULTS: Compared to controls, increased plasminogen activation was observed in the basal ganglia and the cortex of the ischemic hemisphere after 48, 72, and 168 h (affected area of basal ganglia: 44.5+/-21.9, 70.1+/-2.3 and 66.6+/-2.8%, respectively; affected area of cortex: 63.4+/-9.8, 67.7+/-0.7 and 64.0+/-3.7%, respectively). The duration of ischemia had no significant influence on the extent of plasminogen activation. Areas of increased plasminogen activation significantly overlapped with and exceeded areas of MAP 2 loss (P<0.005). DISCUSSION: Permanent focal cerebral ischemia leads to increased plasminogen activation in ischemic regions. This plasminogen activation remains elevated at persistent levels over days. It may contribute to extracellular matrix (ECM) disruption, secondary hemorrhage, and brain edema in subacute stages of ischemic stroke. PMID- 11056181 TI - Effect of P2 purinoceptor antagonists on kainate-induced currents in rat cultured neurons. AB - The action of purinergic antagonists on kainate-induced currents was studied in rat cortical neurons in primary culture using the whole-cell configuration of the patch-clamp technique. The amplitude of the currents induced by kainate in cortical neurons was concentration-dependent (EC(50)=106 microM). Pyridoxal phosphate-6-azophenyll-2',4'-disulphonic acid 4-sodium (PPADS), a P2X antagonist, was ineffective in the reduction of the kainate-induced current in cortical neurons, while 2, 2'-pyridylisatogen (PIT), basilen blue (BB) and suramin, respectively two selective P2Y and a non-selective P2 receptor antagonist, caused a reduction in the amplitude of the current induced by kainate. BB decreased the inward current induced by kainate at all holding potentials and the reduction was dose-dependent (EC(50)=34 microM). The total conductance of the neurons for the kainate-induced current was significantly reduced (P<0.01) and the effect was completely reversible. BB furthermore reduced the kainate-induced current in granule and hippocampal neurons and decreased the amplitude of the alpha-amino-3 hydroxy-5-methyl-4-isoxalepropionic acid (AMPA)-evoked current in cortical neurons. Cholera toxin (ChTx) did not affect the action of BB on the kainate induced currents in cortical neurons and moreover, when guanosine 5'-o-(3 thiotriphosphate) (GTPgammaS) was added to the electrode solution, the kainate induced currents were still reduced by 100 microM BB. The maximal response to kainate decreased in the presence of 20 microM BB without changing its EC(50), indicating a non-competitive mechanism of inhibition. These results demonstrate that preferential P2Y receptor antagonists are able to modulate the kainate and AMPA-induced currents in central neurons, suggesting a potential use of these compounds as neuroprotective agents. PMID- 11056182 TI - Role of spinal alpha(1)-adrenergic mechanisms in the control of lower urinary tract in the rat. AB - The role of spinal alpha(1)-adrenergic mechanisms in the control of urinary bladder function was examined in urethane (1.2 g/kg s.c.) anesthetized and decerebrate unanesthetized female Sprague-Dawley rats (250-320 g). Bladder activity was recorded via a transurethral catheter during continuous infusion (0.21 ml/min) cystometrograms or under isovolumetric conditions. All drugs were administered intrathecally at the L(6)-S(1) segmental level of spinal cord. During cystometrograms, 3 or 30 nmol of phenylephrine (alpha(1)-adrenergic agonist) did not alter bladder activity; whereas 300 nmol increased the intercontraction interval by 98% and pressure threshold for inducing micturition by 115%, but did not change bladder contraction amplitude. A large dose of phenylephrine (3000 nmol) completely blocked reflex voiding and induced overflow incontinence at a high baseline pressure (mean: 33 cmH(2)O; range: 28-42 cmH(2)O). Under isovolumetric conditions, 3-30 nmol of phenylephrine abolished bladder activity for 22-45 min; whereas smaller doses (0.003-0.3 nmol) were inactive. Doxazosin (50 nmol), an alpha(1)-adrenergic antagonist, decreased intercontraction intervals but did not change bladder contraction amplitude during cystometrograms. Under isovolumetric conditions this dose of doxazosin increased bladder contraction frequency and decreased bladder contraction amplitude. Smaller doses (5 or 25 nmol) of doxazosin did not alter bladder activity. These studies suggest that two types of spinal alpha(1)-adrenergic mechanisms are involved in reflex bladder activity: (1) inhibitory control of the frequency of voiding reflexes presumably by regulating afferent processing in the spinal cord and (2) facilitatory modulation of the descending limb of the micturition reflex pathway. PMID- 11056183 TI - The effects of protein kinase C and calmodulin kinase II inhibitors on vestibular compensation in the guinea pig. AB - Previous studies have demonstrated that vestibular compensation, the process of behavioural recovery which occurs following unilateral deafferentation of the vestibular labyrinth (UVD), is correlated with changes in in vitro phosphorylation of various protein substrates in the brainstem vestibular nucleus complex (VNC). The aim of the present study was to investigate the possible causal relationship between protein kinase activity and the induction of the vestibular compensation process, by delivering inhibitors of protein kinase C (PKC) or Ca(2+)/calmodulin-dependent kinase II (CaMKII) into the ipsilateral VNC at the time of the UVD and determining their effects on three static symptoms of UVD, spontaneous nystagmus (SN), yaw head tilt (YHT) and roll head tilt (RHT) in guinea pigs. Infusion of the PKC inhibitor, 3-[1-(3-dimethylaminopropyl)-1H-indol 3-yl]-4-(1H-indol-3-yl)-1H-pyrr ole-2,5-dione, HCl (bisindolylmaleimide I, HCl/GF 109203X, HCl) ('Bis I'), at a concentration of 5 or 50 microM, significantly increased SN frequency at the earliest time points (6 and 8 h post-UVD) compared to vehicle controls and the less selective analogue, 2,3-bis(1H-indol-3-yl)-N methylmaleimide (bisindolylmaleimide V) ('Bis V'). However, the compensation of YHT and RHT was unaffected by the PKC inhibitor. By contrast, the cell-permeable CaMKII inhibitor, myristoylated autocamtide-2 related inhibitory peptide (N-Myr Lys-Lys-Ala-Leu-Arg-Arg-Gln-Glu-Ala-Val-Asp-Ala-Leu-OH) ('myr-AIP') or the cell impermeable analogue, autocamtide-2 related inhibitory peptide (N-Lys-Lys-Ala-Leu Arg-Arg-Cln-Glu-Ala-Val-Asp-Ala-Leu-OH) ('AIP'), failed to alter the compensation of SN, YHT or RHT at any dose compared to vehicle controls. These results implicate PKC-, but not CaMKII-, signal transduction pathways in the initiation of SN compensation in guinea pig. PMID- 11056184 TI - The over-expression of somatostatin in the gerbil entorhinal cortex induced by seizure. AB - In present study, we investigated the immunohistochemical distribution of somatostatin (SRIF) in the hippocampal complex of the Mongolian gerbil and its association with different sequelae of spontaneous seizures, in an effort to identify the roles of SRIF in the self-recovery mechanisms in these animals. In the dentate gyrus and subiculum, SRIF immunoreactive (SRIF(+)) cells were similar in both the seizure resistant and the pre-seizure group of seizure sensitive gerbils. Interestingly, SRIF immunoreactivity was markedly decreased until 12 h postictal. Twenty-four hours after the on-set of seizure, the distribution of SRIF immunoreactivity in these regions had slightly increased. In contrast, in the entorhinal cortex the population of SRIF(+) cells and their density were significantly elevated compared to pre-seizure group 30 min postictal. Twelve hours after the on-set of seizure, however, the population of SRIF(+) cells and their density declined, approximately 70-80% compared to the situation at 30 min postictal. These findings suggest that the enhancement of SRIF expression in gerbil entorhinal cortex may affect tissue excitability and have a role in modulating recurrent excitation following seizures. PMID- 11056185 TI - Is there an optimal age for recovery from motor cortex lesions? I. Behavioral and anatomical sequelae of bilateral motor cortex lesions in rats on postnatal days 1, 10, and in adulthood. AB - Rats were given bilateral lesions of the motor cortex on the day of birth (P1), tenth day of life (P10), or in adulthood. They were trained on several motor tasks (skilled forelimb reaching, beam traversing, tongue extension), general motor activity, and a test of spatial learning (Morris water task). Although all lesion groups were impaired at skilled reaching, the P10 group was less impaired than either of the other two lesion groups. Furthermore, on the other motor tests the P10 group did not differ from controls whereas both P1 and adult groups were impaired. Only the P1 lesion group was impaired at the acquisition of the Morris water task. Anatomical analyses revealed that the P1 and P10 rats had smaller brains than the other two groups as well as having a generalized decrease in cortical thickness. Dendritic analysis of layer III pyramidal cells in the parietal cortex revealed a decrease in apical arbor in the lesion groups and an increase in the basilar arbor of the P1 and adult lesion animals. The P1 and adult operated groups showed an increase in spine density in the basilar dendrites of layer V pyramidal cells. Finally, analysis of the pattern of corticospinal projections revealed that the P1 animals had a markedly wider field of corticospinal projection neurons than any of the other groups. The widespread anatomical changes in all lesion groups versus the relatively better behavioral recovery after P10 lesions suggests that day 10 represents an optimal period for adapting to brain damage and subsequent brain reorganization. PMID- 11056186 TI - Estrogen replacement does not prevent the loss of choline acetyltransferase positive cells in the basal forebrain following either neurochemical or mechanical lesions. AB - Recent studies have shown that estrogen replacement can enhance the functional status of basal forebrain cholinergic neurons. Studies have also shown that estrogen has neuroprotective effects both in vitro and in vivo on a variety of cells and against a variety of insults. The present study examined the ability of estrogen replacement to protect basal forebrain cholinergic neurons from the effects of neurochemical and mechanical injury. Ovariectomized Sprague-Dawley rats received either estrogen replacement or sham surgery, and then received either a unilateral injection of ibotenic acid into the nucleus basalis magnocellularis, or unilateral transection of the fimbria fornix. Cholinergic neurons in the medial septum and nucleus basalis were detected and quantified using immunohistochemical techniques. The data show that neither 3 weeks nor 13 weeks of continuous estrogen replacement prevented the loss of choline acetyltransferase (ChAT)-containing cells in the nucleus basalis following a unilateral injection of ibotenic acid. Likewise, estrogen replacement did not prevent a decrease in ChAT-positive cells detected in the medial septum following unilateral transection of the fimbria fornix. Notably, increased numbers of ChAT positive cells were detected in the contralateral nucleus basalis, and in the ipsilateral and contralateral medial septum, at 2 weeks following a unilateral injection of ibotenic acid into the nucleus basalis; however, these effects were not related to hormone treatment. These data suggest that estrogen replacement does not protect cholinergic neurons in the medial septum and nucleus basalis from the effects of excitotoxic or mechanical injury. PMID- 11056187 TI - Mechanisms underlying H(2)O(2)-mediated inhibition of synaptic transmission in rat hippocampal slices. AB - Hydrogen peroxide (H(2)O(2)) inhibits the population spike (PS) evoked by Schaffer collateral stimulation in hippocampal slices. Proposed mechanisms underlying this effect include generation of hydroxyl radicals (.OH) and inhibition of presynaptic Ca(2+) entry. We have examined these possible mechanisms in rat hippocampal slices. Inhibition of the evoked PS by H(2)O(2) was sharply concentration-dependent: 1.2 mM H(2)O(2) had no effect, whereas 1.5 and 2.0 mM H(2)O(2) reversibly depressed PS amplitude by roughly 80%. The iron chelator, deferoxamine (1 mM), and the endogenous.OH scavenger, ascorbate (400 microM), prevented PS inhibition, confirming.OH involvement. Isoascorbate (400 microM), which unlike ascorbate is not taken up by brain cells, also prevented PS inhibition, indicating an extracellular site of.OH generation or action. We then investigated whether H(2)O(2)-induced PS depression could be overcome by prolonged stimulation, which enhances Ca(2+) entry. During 5-s, 10-Hz trains under control conditions, PS amplitude increased to over 200% during the first three-four pulses, then stabilized. In the presence of H(2)O(2), PS amplitude was initially depressed, but began to recover after 2.5 s of stimulation, finally reaching 80% of the control maximum. In companion experiments, we assessed the effect of H(2)O(2) on presynaptic Ca(2+) entry by monitoring extracellular Ca(2+) concentration ([Ca(2+)](o)) during train stimulation in the presence of postsynaptic receptor blockers. Evoked [Ca(2+)](o) shifts were apparently unaltered by H(2)O(2), suggesting a lack of effect on Ca(2+) entry. Taken together, these findings suggest new ways in which reactive oxygen species (ROS) might act as signaling agents, specifically as modulators of synaptic transmission. PMID- 11056188 TI - Intracerebroventricular injection of senktide-induced Fos expression in vasopressin-containing hypothalamic neurons in the rat. AB - Intracerebroventricular injection of senktide, a selective agonist for neurokinin B receptor (NK3), induced Fos expression in many neurons of the rat hypothalamus. Fos-positive neurons were predominantly present in the supraoptic and paraventricular hypothalamic nuclei, and some of them were seen in the lateral preoptic area, lateral hypothalamic area, arcuate nucleus, perifornical region, posterior hypothalamic area, circular nucleus, and along relatively large blood vessels (lateral hypothalamic perivascular nucleus) in the anterior hypothalamus. A double labeling study was performed to examine if vasopressin-containing neurons in the hypothalamus could be activated by the treatment. Neurons with both Fos-like immunoreactivity (-LI) and vasopressin-LI were found in the paraventricular nucleus, supraoptic nucleus, circular nucleus and lateral hypothalamic perivascular nucleus. In the supraoptic nucleus, about 87% of vasopressin-containing neurons exhibited Fos-LI, which corresponded to about 64% of Fos-positive neurons in the nucleus. In the paraventricular nucleus, about 80% of vasopressin-like immunoreactive neurons exhibited Fos-LI, which constituted about 51% of the total population of Fos-positive neurons in the region. The results suggest that NK3 receptor may be involved in the modulation of release of vasopressin from the hypothalamus in the rat. PMID- 11056189 TI - Dual control of the brainstem on the spindle oscillation in humans. AB - In human subjects, the excitability change of the brainstem was investigated over the course of the spindle oscillation. The investigation was carried out by a sequential analysis of brainstem auditory evoked potentials (BAEPs) with reference to one sequence of spindle oscillation. The method was based on the characteristics of BAEPs, i.e. far-field evoked potential. The brainstem revealed two types of excitability change: one in the lower ventral brainstem (wave-III components), and the other in the upper dorsal brainstem (wave-V components). The excitability in the dorsal brainstem showed an oscillation with one cycle period of about 1.5 s, whereas in the ventral brainstem, the excitability showed a long range biphasic (decaying-growing) fluctuation. Both excitability changes in the brainstem preceded the spindle oscillation, and the phase was reversed during the emerging period of spindle oscillation. The results suggest a primary triggering mechanism of the brainstem for the spindle oscillation, which is independent of preceding cortical drives (K-complexes) upon the thalamus. The difference of the excitability change between the spindle oscillation and the paroxysmal discharge (spike-and-wave complex) was also discussed. PMID- 11056190 TI - Auditory noise can prevent increased extracellular acetylcholine levels in the hippocampus in response to aversive stimulation. AB - The intent of this study was to investigate neurochemical and behavioural effects of aversive stimulation and the impact of auditory background noise. Using in vivo microdialysis, hippocampal acetylcholine was extracted and subjected to HPLC analysis while male Wistar rats were exposed to aversive stimulation similar to that used in conventional procedures for aversive conditioning. Three groups of animals were used. Animals in the first group were exposed to a single tone/footshock pairing followed by a tone alone 2 h later. Animals in the second group served as controls and were only exposed to the tone without shock. A third group was exposed to the same tone/shock pairing and tone as the first group while being exposed to constant background noise during the whole experiment. The results showed, that the tone/shock combination led to pronounced behavioral and cholinergic activation. In contrast, exposure to background noise prevented the increase in hippocampal ACh levels to tone/shock stimulation. The unconditioned behavioural response, however, was not prevented suggesting that hippocampal ACh is not a necessary correlate of behavioural activation or arousal. A second experiment intended to investigate the effects of background noise in a shuttle box avoidance learning paradigm where rats were trained to avoid an aversive footshock, which was signalled by a tone. There, one group of rats was exposed to background noise during avoidance learning, and the other group was not exposed to noise. Whereas both groups learned to avoid the shock to some degree over training, the noise exposed animals did not show improvement in escape performance over the course of training, indicating that the noise hindered development of an adaptive response to the shock. In summary, our data indicate that background noise can prevent increased extracellular hippocampal ACh levels in response to an aversive stimulus, and can also lead to deficits in learning to escape from shock. PMID- 11056191 TI - Effect of experimental pain from trigeminal muscle and skin on motor cortex excitability in humans. AB - The pathophysiology of many orofacial pain syndromes is still unclear. We investigated the effect of tonic muscle and skin pain on the excitability of the trigeminal motor pathways using transcranial magnetic stimulation (TMS). Motor evoked potentials (MEPs) were recorded in the masseter surface electromyogram (EMG). Magnetic pulses were delivered with a large coil at intensities 1.1 and 1.5 times the motor threshold, and for each intensity, MEPs were recorded at three different clenching levels: 15, 30 and 45% of maximum voluntary contraction (MVC). Baseline, pain and post-baseline recordings were compared in two sessions. Firstly, muscle pain was induced by infusion of hypertonic saline (5.8%) into the left masseter. Secondly, skin pain was induced by topical application of capsaicin (5%) on the left cheek. Muscle and skin pain did not induce significant effects on the amplitude or latency of the MEPs (ANOVAs: P>0.50). In both sessions, the amplitude of the MEPs increased with the increase of the clenching level and stimulus intensity (P<0.0001; P<0.005) whereas the latency was not significantly changed (P>0.05; P=0.11). Muscle pain was associated with an increase in the pre-stimulus EMG activity on the non-painful side compared with baseline (P<0.01), which could be due to compensatory changes in the activation of the painful muscle. The need for voluntary contraction to evoke MEPs in the masseter muscles and compensatory mechanisms both at the brainstem and cortical level might explain the lack of detectable modulation of MEPs. Nonetheless, the present findings did not support the so-called 'vicious cycle' between pain - central hyperexcitability - muscle hyperactivity. PMID- 11056192 TI - Crucial role of kainate receptors in mediating striatal kainate injection-induced decrease in acetylcholine M(1) receptor binding in rat forebrain. AB - We investigated the roles of kainate-, alpha-amino-3-hydroxy-5-methylisoxazol-4 propionate (AMPA)- and N-methyl-D-aspartate (NMDA)-receptors in mediating striatal kainate injection-induced decrease in the binding of acetylcholine M(1) receptors in rat forebrain. After unilateral intrastriatal injection of kainate (4 nmol), the bindings of [3H]kainate (10 nM), [3H]MK-801 (4 nM) and [3H]pirenzepine (4 nM) to the rat ipsilateral forebrain membranes declined, reaching the lowest on day 2 to 4 and recovering on day 8. Saturation binding studies, performed on day 2 post-injection, showed that kainate (1, 2, 4 nmol) dose-dependently decreased B(max) and K(d) of the three ligands. (+)-5-Methyl-10, 11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine (MK-801), a selective NMDA receptor channel blocker, antagonised (from a dose of 4 nmol) kainate-induced decreases in the bindings of [3H]kainate (up to approximately 20%), [3H]MK-801 (up to approximately 90%) and [3H]pirenzepine (up to approximately 70%). In contrast, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), a selective non-NMDA receptor antagonist, almost completely abolished (from a dose of 12 nmol) kainate induced decreases in the bindings of all the three ligands (up to approximately 95-98%). Cyclothiazide, a selective potentiator that enhances AMPA receptor mediated responses, significantly enhanced (from a dose of 4 nmol) kainate induced decrease in the binding of [3H]kainate but not that of [3H]pirenzepine or [3H]MK-801. In summary, these results indicate that striatal kainate injection induced decrease in the binding of acetylcholine M(1) receptors in rat forebrain is dependent on activation of kainate receptors and, to a certain extent, a consequent involvement of NMDA receptors. These and previous studies provide some evidence showing that kainate receptors might play a crucial role in regulating excitatory amino acids (EAA)-modulated cholinergic neurotransmission in the central nervous system (CNS). PMID- 11056193 TI - Chronic cocaine differentially affects diazepam's anxiolytic and anticonvulsant actions. Relationship to GABA(A) receptor subunit expression. AB - Benzodiazepines are used to treat the anxiety associated with cocaine withdrawal, as well as cocaine-induced seizures. Since cocaine exposure was shown to affect BZ binding density, abuse liability, subjective hypnotic actions and seizure susceptibility, we assessed whether chronic cocaine alters diazepam's anxiolytic and anticonvulsant actions. Changes in GABA(A) receptor subunit protein expression were also assessed as they may relate to BZ activity at the receptor. Male Sprague-Dawley rats were injected with cocaine-HCl (15 mg/kg, i.p.) or saline once daily for 14 days. One day after the last injection, DZP (1 mg/kg i.p.) significantly increased time spent on and entries into open arms of an elevated plus maze in both saline- and cocaine-treated groups, yet the effect was greater in cocaine-treated rats. Eight days after cessation of treatment DZP did not have a significant anxiolytic effect in either group. To assess the effect of cocaine on DZP's anticonvulsant actions, PTZ was infused at a constant rate via the lateral tail vein and clonus onset was recorded in the presence and absence of DZP (5 mg/kg, i.p). DZP significantly elevated seizure threshold in both groups of rats. Chronic cocaine also had no effect on the beta-CCM seizure threshold. Quantitative immunohistochemistry of GABA(A) receptor subunit protein demonstrated significant regulation of alpha2 (-10%) and beta3 (+9%) subunits in the hippocampal dentate gyrus and CA1 regions, respectively. Small changes in GABAR subunit expression in specific brain areas may relate to DZP's enhanced anxiolytic effectiveness whereas it's anticonvulsant actions likely remain intact following cocaine administration. PMID- 11056194 TI - Regional differences in brain monoamine oxidase subtypes in an animal model of geriatric depression: effects of olfactory bulbectomy in young versus aged rats. AB - Geriatric depression is often associated dysregulation of the hypothalamus pituitary-adrenal (HPA) axis, and with poor responsiveness to antidepressants that work through inhibition of monoamine reuptake; accordingly, it has been suggested that MAO inhibitors may represent a therapeutic alternative in this group. In the current study, we evaluated expression of MAO subtypes in brain regions of young and aged rats subjected to olfactory bulbectomy (OBX), a procedure that reproduces many of the biochemical and functional changes associated with human depression. Activities of both MAO A and B were elevated in aged rats as compared to young rats in most regions, but not in the midbrain, and the OBX lesion failed to produce any change in this pattern. These results stand in contrast to the differential effects of glucocorticoids, which reduce brain MAO in young animals but induce activity in aged rats. Our results support the view that the aged brain possesses biochemical characteristics that distinguish its monoamine biochemistry from that of young brain, and that these distinctions may work in conjunction with HPA axis dysregulation to influence the etiology and therapy of geriatric depression. The use of appropriate animal models for depression and for disruption of HPA axis function can allow for the testing of potential human biomarkers (such as platelet MAO) that may serve to predict treatment outcome. PMID- 11056195 TI - Choline and acetylcholine have similar kinetic properties of activation and desensitization on the alpha7 nicotinic receptors in rat hippocampal neurons. AB - The alpha7-type nicotinic acetylcholine receptor (nAChR) was recently found to be both fully activated and desensitized by choline, in addition to ACh. In order to understand the combined effects of the two agonists on alpha7 nAChR-mediated neuronal signaling, the kinetics of the receptor-channel's interaction with ACh and choline was examined. To this end, whole-cell and single-channel currents evoked by fast-switching pulses of the agonists were recorded in rat hippocampal neurons in culture. Currents evoked by equieffective concentrations of choline and ACh were very similar, except that choline-evoked currents decayed more quickly to the baseline after removal of the agonist, and that recovery from desensitization was faster with choline. The conductance of channels activated by choline and ACh was 91.5+/-8.5 and 82.9+/-11.6 pS, respectively. The mean apparent channel open times were close to 100 micros, with both agonists. After a 4-s exposure to concentrations up to 80 microM ACh or 600 microM choline, the extent of desensitization and the cumulative charge flow carried by the channels increased in the same proportion, until reaching a maximum. At higher concentrations of either agonist, the cumulative charge started decreasing with concentration, reflecting further desensitization. Kinetic modeling suggested that alpha7 nAChRs have at least two non-equivalent paths to desensitized states, and that choline dissociates faster than ACh from the receptor. Our results established that the main difference between choline and ACh is of affinity, and support the concept that the switching of endogenous agonist may change the desensitization-resensitization dynamics of alpha7 nAChRs. PMID- 11056196 TI - Cellular dynamics of corneal wound re-epithelialization in the rat. III. Mitotic activity. AB - The number and distribution of mitotic epithelial cells in the ocular surface during homeostasis and in response to abrasion of the mammalian cornea were determined. Normal rats and those receiving a 3 mm diameter centrally located epithelial defect, received an intraperitoneal injection of colchicine 6 h prior to sacrifice. Mitosis in the basal epithelium during homeostasis was comparable in magnitude across the ocular surface epithelium, with the exception of a few mitotic figures in the midline. Thirty percent of the mitotic figures were in the basal layer (layer 1), and 70% were in layer 2; the cells in layer 2 were often noted to retain connection to the basal lamina by cytoplasmic stalks. Mitosis was rarely noted in the regenerating epithelium. However, summation of M phase cells in both the basal and suprabasal epithelium adjacent to the wound showed increases of 3- and 5-fold at 30 and 36 h after abrasion, respectively, from levels at homeostasis and the time of injury. In striking contrast to homeostatic epithelium, 80% of the mitotic cells were located in layer 1 of the corneal epithelium, with normal distribution observed by 72 h. Mitosis in the limbus and conjunctiva was increased 3-fold at 30 h and 24 h, respectively, from values at homeostasis and the time of debridement. These results, using rigorous statistical analysis and precise topographic assessment, showed that mitosis is not impeded - but rather often accelerated - following denuding of the corneal epithelium and that the spatial distribution of mitotic cells is correlated with wounding. The data revealed that re-epithelialization of the corneal epithelium is not dependent on mitosis in the regenerating epithelium, but rather in the adjacent unwounded epithelium of the cornea, with most cells being located in the basal layer until re-epithelialization is completed. Mitotic cells in the limbus and conjunctiva may be related to replenishment of ocular surface epithelial cells used in the repair process rather than directly supplying the abraded surface. PMID- 11056197 TI - Pituitary adenylate cyclase-activating polypeptide innervation of the mudpuppy cardiac ganglion. AB - The presence and potential origin of the neuropeptide pituitary adenylate cyclase activating polypeptide (PACAP) was determined in cardiac ganglia of the mudpuppy, Necturus maculosus. Although PACAP has been implicated in the regulation of cardiac function in several mammalian species, the presence of this peptide in the autonomic nervous system (ANS) of other species is unclear. Thus, this study is the first to characterize this highly conserved peptide in the ANS of a non mammalian species. PACAP-immunoreactivity was observed in nerve fibers throughout the mudpuppy cardiac ganglia and often was co-localized with the sensory neuropeptides substance P and calcitonin gene-related peptide. Removal of all extrinsic inputs to the ganglia by organ culture eliminated PACAP immunoreactivity in the cardiac ganglia, whereas bilateral vagotomies only partially reduced PACAP-labeling. PACAP-immunoreactive neurons were observed in both high thoracic dorsal root ganglia and in vagal sensory ganglia. While no PACAP-positive neurons were observed in caudal medulla brainstem regions, PACAP containing nerve fibers were found in the region of the nucleus solitarius. These results suggest that, in the mudpuppy, PACAP is found primarily in visceral afferent fibers, originating from cells in either the dorsal root ganglia or vagal sensory ganglia. Based on their anatomic localization, these afferent fibers may function to transmit important sensory information to cardiovascular centers in the brain as well as serving as local reflex inputs to modulate postganglionic parasympathetic output within the cardiac ganglion itself. PMID- 11056198 TI - Region-specific attenuation of a trypsin-like protease in substantia nigra following dopaminergic neurotoxicity by 1-methyl-4-phenyl-1,2, 3,6 tetrahydropyridine. AB - We analysed apoptosis, caspase-1 and -3, and trypsin-like protease activity in the nigrostriatal pathway during 1-methyl-4-phenyl-1,2,3, 6-tetrahydropyridine (MPTP)-induced neurotoxicity. MPTP injected (30 mg/kg, i.p., twice, 16 h apart) mice were sacrificed on 1, 2 and 7 days. DNA extracted from nucleus caudatus putamen (NCP) and substantia nigra (SN) was subjected to agarose gel electrophoresis. Typical apoptotic-like DNA cleavage was absent in SN or NCP after this dose of MPTP. A trypsin-like protease activity was significantly decreased in SN and not in NCP. While caspase-3 activity in the whole brain was increased significantly, caspase-1 activity was unaffected. Striatal dopamine content was decreased to 75% by 7 days. The absence of typical DNA 'ladder' when there was severe striatal dopamine depletion suggests that in vivo MPTP-mediated dopaminergic neurotoxicity may not involve apoptotic cell death, and explains why in mice MPTP-induced dopamine depletion is transient. The region-specific decrease in trypsin-like protease activity and absence of caspase-3 activation in SN signify the importance of trypsin-like protease in the regulation of apoptosis in MPTP-neurotoxicity in mice. PMID- 11056199 TI - Nicotine-induced locomotor activity is increased by preexposure of rats to prenatal stress. AB - Genetic factors are believed to play a predominant role in the individual differences observed in behavioral and pharmacological responses to drugs of abuse. An increasing literature indicates, however, that epigenetic factors can be involved as well. In this report we examined whether developmental changes induced by prenatal stress could alter the way animals respond to the psychostimulant effect of nicotine when adults. The results show that nicotine induces a dose-dependent increase of locomotor activity in both groups, and that prenatally-stressed animals exhibit a higher behavioral response at all doses. This study emphasizes the importance of early environment in the later development of drug-related behavior. PMID- 11056200 TI - Acetylcholine sensitivity in sensory neurons dissociated from the cat petrosal ganglion. AB - The petrosal ganglia contain the somata of the sensory fibers of the glossopharyngeal nerves, innervating structures of the tongue, pharynx, carotid sinus and carotid body. Petrosal ganglia were excised from adult cats and their neurons were dissociated and kept in tissue culture for 7-12 days. Intracellular recordings were obtained through conventional microelectrodes. In response to depolarizing pulses, most cells (41/60) presented a 'hump' in the falling phase of their action potentials (H-type), while the remaining neurons lack such hump (F-type). The two types of cells had no differences in resting membrane potential or action potential amplitude. Acetylcholine (ACh) applied locally elicited responses in nearly two thirds of both H-type and F-type neurons tested. Most H type neurons (17/19) responded with a slow long lasting depolarization, while the remaining (2) did so by generating spikes. In contrast, half of F-type neurons (6/12) responded with one or more spikes and the other half only with a slow depolarization. These results indicate that ACh receptors are present in the soma of many petrosal ganglion neurons subjected to tissue culture, thus supporting the idea that - under normal conditions - their peripheral sensory processes may be excited by ACh. PMID- 11056201 TI - Expression of microglial response factor-1 in microglia and macrophages following cerebral ischemia in the rat. AB - Microglial response factor-1 is a newly isolated microglial gene, which encodes a Ca(2+) binding protein MRF-1 expressed in microglia and macrophages. We induced 1 h of focal cerebral ischemia or 10 min of global cerebral ischemia in the rat, and investigated the expression of MRF-1 immunoreactivity following ischemia. MRF 1 was present in resting microglia and was upregulated in response to microglial activation. MRF-1 was localized to all the cells of the mononuclear phagocyte system (microglia, monocytes, and perivascular cells) that appeared in the ischemic brain. PMID- 11056202 TI - Effects of brain oxygenation on metabolic, hemodynamic, ionic and electrical responses to spreading depression in the rat. AB - The effect of cortical spreading depression (CSD) on oxygen demand (extracellular K(+)), oxygen supply (cerebral blood flow - CBF) and oxygen balance (mitochondrial NADH) was studied by a special multiprobe assembly (MPA), during hypoxia and partial ischemia. The MPA was constructed and applied to monitor the CSD wave from its front line until complete recovery, continuously and simultaneously. CSD under hypoxia or partial ischemia led to an initial increase in NADH levels and a further decrease in CBF during the first phase of the CSD wave, indicating a decrease of tissue capability to compensate for an increase in oxygen demand. Furthermore, the special design of the MPA enabled identifying the close interrelation between oxygen demand, supply and balance during CSD propagation. In conclusion, brain oxygenation was shown to have a clear effect on tissue responses to CSD. PMID- 11056203 TI - Activation of ATP receptor increases the cytosolic Ca(2+) concentration in nucleus accumbens neurons of rat brain. AB - ATP increased the cytosolic Ca(2+) concentration ([Ca](i)) in nucleus accumbens neurons acutely dissociated from rat brain. The ATP response was dependent on external Ca(2+) and Na(+), and was blocked by voltage-dependent Ca(2+) channel blockers. The results suggest that the ATP-induced depolarization increases Ca(2+) influx resulting in the increase in [Ca](i). PMID- 11056204 TI - Schwann cell-induced loss of synapses in the central nervous system. AB - Synaptophysin immunostaining of areas of spinal gray matter occupied by radiation induced intraspinal Schwann cells revealed a loss of immunoreactivity from the neuropil. In contrast, synaptophysin immunoreactivity was preserved on the somata and proximal dendrites of motor neurons. The present study extended these observations to the ultrastructural level and confirmed the absence not only of synapses but also of astrocytes and small- and medium-sized dendrites. These neural elements were abundant and appropriately organized in contiguous areas of irradiated neuropil not occupied by Schwann cells. PMID- 11056205 TI - Effects of the Na(+)/H(+) exchanger monensin on intracellular pH in astroglia. AB - Stimulation of astroglial glucose utilization by the Na(+)/H(+) exchanger monensin is only partially blocked by ouabain. The present studies show that monensin also raises intracellular pH in astroglia. Because increased pH stimulates phosphofructokinase activity, the ouabain-insensitive portion of the stimulation of cerebral glucose utilization (CMR(glc)) appears to be due to stimulation of glycolysis by intracellular alkalinization. PMID- 11056206 TI - Cortical cholinergic activity is related to the novelty of the stimulus. AB - A number of studies have related cholinergic activity to the mediation of learning and memory. However, the acetylcholine (ACh) participation has been recently implicated in the early stages of memory formation but not during retrieval. The aim of the present study is to evaluate ACh release in the insular cortex (IC) during presentation of different taste stimuli and during their re exposition by means of the free-moving microdialysis technique. We evaluated the changes in ACh release when a novel taste, saccharin or quinine was presented to the rat and after several presentations of saccharin. Unilateral microdialysis was performed in the IC 1 h before and 1 h after the presentation of: (1) a familiar stimulus (water), (2) a novel taste (quinine), (3) another novel taste (saccharin), (4) a second presentation, (5) a third presentation, and (6) a fourth presentation of saccharin. The volume consumed by the animals was registered as a behavioral parameter. The ACh levels from the microdialysis fractions were analyzed by an HPLC-ED system. Biochemical results showed a significant increment in the cortical ACh release induced by a novel stimulus compared with the release observed during the presentation of a familiar stimulus. The ACh release observed after several presentations of the stimuli decreased to the same levels as those produced by the familiar taste, indicating an inverse relationship between familiarity and cortical ACh release. These results suggest that the cholinergic system plays an important role in the identification and characterization of different kinds of stimuli. PMID- 11056207 TI - Expression of estrogen receptors in the dorsal root ganglia of the chick embryo. AB - Estrogen receptors (ER) are widely distributed in the central nervous system (CNS). Recent studies, to date in rat only, have shown that ER are also expressed in neurons of the dorsal root ganglia (DRG) where they appear to have functional roles. However, no data yet exists about estrogen receptors in the embryonic DRG. In the present study, immunocytochemical staining for ER in the DRG of chick embryos from day 6.5 to 18.5 (Hamburger and Hamilton St. 30-45) of incubation was performed. ER+ cells were first consistently observed at day 8.5 (St. 34), more concentrated in the ventral-lateral portion of the DRG. From day 8.5 to 12.5 (St. 38), the density of ER+ cells and the staining intensity increased, with no obvious changes from day (E) 12.5 to 18.5. Although ER is detected mainly in the cytoplasm of embryonic DRG neurones, ER+ cells with nuclear staining are sometimes observed and gradually increase in number during development. ER immunoreactivity in the DRG at cervical, thoracic and lumbo-sacral levels is similar and no obvious differences in staining were observed between male and female embryos. ER+ neurons are also present in the sympathetic ganglia from E8.5 and some primary spinal motoneurons are ER+ beginning at E14.5. The results suggest that estrogen may play a role in the embryonic development of the DRG. PMID- 11056208 TI - Functional plasticity in extrastriate visual cortex following neonatal visual cortex damage and monocular enucleation. AB - Neonatal lesions of primary visual cortex (areas 17, 18 and 19; VC) in cats lead to significant changes in the organization of visual pathways, including severe retrograde degeneration of retinal ganglion cells of the X/beta class. Cells in posteromedial lateral suprasylvian (PMLS) cortex display plasticity in that they develop normal receptive-field properties despite these changes, but they do not acquire the response properties of striate neurons that were damaged (e.g., high spatial-frequency tuning, low contrast threshold). One possibility is that the loss of X-pathway information, which is thought to underlie striate cortical properties in normal animals, precludes the acquisition of these responses by cells in remaining brain areas following neonatal VC damage. Previously, we have shown that monocular enucleation at the time of VC lesion prevents the X-/beta cell loss in the remaining eye. The purpose of the present study was to determine whether this sparing of retinal X-cells leads to the development of striate-like response properties in PMLS cortex. We recorded the responses of PMLS neurons to visual stimuli to assess spatial-frequency tuning, spatial resolution, and contrast threshold. Results indicated that some PMLS cells in animals with a neonatal VC lesion and monocular enucleation displayed a preference for higher spatial frequencies, had higher spatial resolution, and had lower contrast thresholds than PMLS cells in cats with VC lesion alone. Taken together, these results suggest that preserving X-pathway input during this critical period leads to the addition of some X-like properties to PMLS visual responses. PMID- 11056209 TI - Light-induced zif268 expression is dependent on noradrenergic input in rat visual cortex. AB - In the present paper we investigated the role of the noradrenergic projection from the locus coeruleus on the expression of the immediate early gene zif268 in the visual cortex of rats exposed to ambient light stimulation. Local administrations of 6-hydroxydopamine (6-OHDA), a specific toxin directed against the catecholaminergic system, were performed in the locus coeruleus prior to visual stimulation. Animals were stimulated for 2 h by ambient light, after a 2 week dark adaptation period. Sham-operated controls displayed a massive increase in the number of zif268 positive cells after light stimulation. To the contrary, lesioned animals demonstrated a dramatic reduction in the number of zif268 positive nuclei across all cortical layers. A few scattered immunopositive nuclei were identified in cortical layer IV, however, this region also underwent a significant reduction in the number of zif268 immunopositive nuclei. Our results indicate that the noradrenergic system plays an important role in the expression of zif268 in the visual cortex of rats exposed to ambient light after dark isolation. PMID- 11056210 TI - Different stimulatory opioid effects on intracellular Ca(2+) in SH-SY5Y cells. AB - Present study revealed the stimulatory effects of delta opioid receptor on intracellular Ca(2+) concentration ([Ca(2+)](i)) in SH-SY5Y cells. Fura-2 based single cell fluorescence ratio (F345/F380) was used to monitor the fluctuation of [Ca(2+)](i). Application of the selective delta-opioid receptor agonist alone, [D Pen(2,5)]-enkephalin (DPDPE), hardly had any effects on cells cultivated for 3-10 days. However, after the cells had been pre-stimulated with cholinoceptor agonist, carbachol, variable calcium elevation was found in 59% of the cultures. The response was naltridole-reversible and dose-dependent, and was abolished completely by thapsigargin (TG) treatment but not by administration of CdCl(2) or 0-Ca(2+) bath solutions. DPDPE-mediated [Ca(2+)](i) elevation was abolished by pertussis toxin (PTX) pretreatment but not cholera toxin (CTX), indicating coupling via G proteins of G(i)/G(o) subfamily. In 17.5% of the responding cells, biphase response was found which may be due to both the stimulatory and the inhibitory effects of opioid. On the other hand, in acutely dissociated cells, DPPDE alone induced [Ca(2+)](i) increase in 50% of the cultures. The probability and the amplitude of the elevation were decreased considerably by application of nifedipine or 0-Ca(2+) bath solution and was little affected by application of TG. DPDPE activated [Ca(2+)](i) increase via a PTX-insensitive and CTX-sensitive pathway suggesting coupling through G(s) subunit. All these indicated the opioid modulated the intracellular Ca(2+) regulation system through different pathways. SH-SY5Y cell line might be a suitable model for the investigation of the complex mechanism which underlies opioid function. PMID- 11056211 TI - Inhibition of interleukin-6 signaling by galiellalactone. AB - A search for inhibitors of the IL-6-mediated signal transduction in HepG2 cells using secreted alkaline phosphatase (SEAP) as reporter gene resulted in the isolation of galiellalactone (1) from fermentations of the ascomycete strain A111 95. Galiellalactone inhibits the IL-6-induced SEAP expression with IC(50) values of 250-500 nM by blocking the binding of the activated Stat3 dimers to their DNA binding sites without inhibiting the tyrosine and serine phosphorylation of the Stat3 transcription factor. Due to its selective activity, galiellalactone may serve as a lead compound for the development of new therapeutic agents for diseases originating from the inappropriate expression of IL-6 and as molecular tool to dissect the JAK/STAT pathways. PMID- 11056212 TI - Transgenic expression of cecropin B, an antibacterial peptide from Bombyx mori, confers enhanced resistance to bacterial leaf blight in rice. AB - The short persistence of cecropin B peptide in plants, due to post-translational degradation, is a serious impediment in its effective utilization for developing bacterial resistance transgenic plants. Two DNA constructs encoding the full length precursor of cecropin B peptide and the mature sequence of cecropin B peptide preceded by a signal peptide derived from rice chitinase gene were transformed in rice. The differences in the transcriptional levels in independent transgenic lines showed moderate to high expression of cecropin B gene that correlated well with the differences in cecropin B accumulation observed by Western blot analysis. The development of lesions resulting from infection by Xanthomonas oryzae pv. oryzae was significantly confined in the infected leaflet of transgenic lines, when compared with the control plants. PMID- 11056213 TI - Molecular evidence of a unique lipoamide dehydrogenase in plastids: analysis of plastidic lipoamide dehydrogenase from Arabidopsis thaliana. AB - Lipoamide dehydrogenase is a subunit of the alpha-ketoacid dehydrogenases and the glycine decarboxylase complex in mitochondria, and the pyruvate dehydrogenase complex in plastids. We report here the unexpected finding of two plastidic isoforms of lipoamide dehydrogenase from Arabidopsis thaliana that are different from the mitochondrial form of the enzyme. The cDNA clones were confirmed by sequence alignment analysis and their location verified by chloroplast import assay. They are single copy genes that appear to be expressed in parallel in different tissues with highest level in developing siliques. Phylogenetic analysis gives further exemplary evidence for the plastidic evolution derived from cyanobacteria. PMID- 11056214 TI - The MAT1 cyclin-dependent kinase-activating kinase (CAK) assembly/targeting factor interacts physically with the MCM7 DNA licensing factor. AB - MAT1 (menage a trois1) functions as an assembly/targeting factor of CAK (cyclin dependent kinase-activating kinase). In a search for MAT1-interacting proteins using yeast two-hybrid system, MCM7 (minichromosome maintenance 7), a member of a family of DNA licensing factors, was identified. The physical interaction between MAT1 and MCM7 was confirmed in vivo in yeast cells and verified with in vitro protein binding assays. Further studies showed the RING-finger motif of MAT1 is not required for the interaction with MCM7, while the C-terminal domain of MAT1 is indispensable. Immunoprecipitation of MCM7 in human osteosarcoma MG63 cells demonstrated that MCM7 associates with the CAK complex in vivo. PMID- 11056215 TI - An acidic amino acid cluster regulates the nucleolar localization and ribosome assembly of human ribosomal protein L22. AB - The control of human ribosomal protein L22 (rpL22) to enter into the nucleolus and its ability to be assembled into the ribosome is regulated by its sequence. The nuclear import of rpL22 depends on a classical nuclear localization signal of four lysines at positions 13-16. RpL22 normally enters the nucleolus via a compulsory sequence of KKYLKK (I-domain, positions 88-93). An acidic residue cluster at the C-terminal end (C-domain) plays a nuclear retention role. The retention is concealed by the N-domain (positions 1-9) which weakly interacts with the C-domain as demonstrated in the yeast two-hybrid system. Once it reaches the nucleolus, the question of whether rpL22 is assembled into the ribosome depends upon the presence of the N-domain. This suggests that the N-domain, on dissociation from its interaction with the C-domain, binds to a specific region of the 28S rRNA for ribosome assembly. PMID- 11056216 TI - The N-terminal domain of the Caulobacter crescentus CgtA protein does not function as a guanine nucleotide exchange factor. AB - The Caulobacter crescentus GTP binding protein CgtA is a member of the Obg/GTP1 subfamily of monomeric GTP binding proteins. In vitro, CgtA displays moderate affinity for both GDP and GTP, and rapid exchange rate constants for either nucleotide. One possible explanation for the observed rapid guanine nucleotide exchange [corrected] rates is that CgtA is a bimodal protein with a C-terminal GTP binding domain and an N-terminal GEF domain. In this study we demonstrate that although the N-terminus of CgtA is required for function in vivo, this domain plays no significant role in the guanine nucleotide binding, exchange or GTPase activity. PMID- 11056217 TI - Effect of genomic and subgenomic leader sequences of potato leafroll virus on gene expression. AB - The effect of the genomic and subgenomic leader sequence of potato leafroll polerovirus on the efficiency of translation of the downstream located genes has been studied. The results obtained in vitro and in vivo indicate that neither leader sequence functions as translational enhancer, a generally important feature of leader sequences. Deletion analyses demonstrated that both leader sequences not only decrease translation of the downstream located genes but also alter the ratio of the synthesized proteins. A correlation between the in vitro and in vivo results can be established in the case of the subgenomic leader sequence. PMID- 11056218 TI - Down-regulation of uncoupling protein-3 and -2 by thiazolidinediones in C2C12 myotubes. AB - Uncoupling proteins (UCPs) are mitochondrial membrane proton transporters that uncouple respiration from oxidative phosphorylation by dissipating the proton gradient across the membrane. We studied the direct effect of several peroxisome proliferator-activated receptor (PPAR) ligands on UCP-3 and UCP-2 mRNA expression in C2C12 myotubes for 24 h. In the absence of exogenous fatty acids, treatment of C2C12 cells with a selective PPARalpha activator (Wy-14,643) or a non-selective PPAR activator (bezafibrate) did not affect the expression of UCP-3 mRNA levels, whereas UCP-2 expression was slightly increased. In contrast, troglitazone, a thiazolidinedione which selectively activates PPARgamma, strongly decreased UCP-3 and UCP-2 mRNA levels. Another thiazolidinedione, ciglitazone, had the same effect, but to a lower extent, suggesting that PPARgamma activation is involved. Further, the presence of 0.5 mM oleic acid strongly increased UCP-3 mRNA levels and troglitazone addition failed to block the effect of this fatty acid. The drop in UCP expression after thiazolidinedione treatment correlated well with a reduction in PPARalpha mRNA levels produced by this drug, linking the reduction in PPARalpha mRNA levels with the down-regulation of UCP mRNA in C2C12 myotubes after thiazolidinedione treatment. PMID- 11056219 TI - Structural and functional similarities between HIV-1 reverse transcriptase and the Escherichia coli RNA polymerase beta' subunit. AB - Four monoclonal antibodies (MAbs) recognizing HIV-1 reverse transcriptase (RT) were shown here to cross-react with the beta' subunit of Escherichia coli RNA polymerase (RNAP). The anti-RT MAbs bind to a peptide comprising residues 294-305 of the RT amino acid sequence. Computer analyses revealed sequence similarity between this peptide and two regions of the RNAP beta' subunit. MAb-binding studies using RT mutants suggested that the epitope is located to amino acids 652 663 of the beta' sequence. One of the MAbs which inhibited the polymerase activity of RT also mediated a dose dependent inhibition of the RNAP activity. PMID- 11056220 TI - Srb7p is essential for the activation of a subset of genes. AB - The mediator complex in the RNA polymerase II holoenzyme is known to be involved in transcriptional activation. The role of the essential mediator component Srb7p has been difficult to investigate, since no conditional lethal allele has been available to date. While the expression of Srb7p under the control of a repressible promoter is not sufficient to reduce the level of Srb7p beneath the threshold for survival, we have been able to isolate a clone termed ts16 which confers a temperature sensitive phenotype. ts16 contains an insertion mutation that requires translational frameshifting for correct expression of Srb7p, leading to extremely low protein levels. Strains bearing the ts16 construct show mild defects in the transcription of constitutive genes like TDH1 but severely affect activated transcription, e.g. of the GAL1 gene. In contrast, CUP1, which is also independent of other holoenzyme components, is not affected by ts16. PMID- 11056221 TI - Inactivation of a MAPK-like protein kinase and activation of a MBP kinase in germinating barley embryos. AB - We provide evidence for involvement of two different 45 kDa protein kinases in rehydration and germination of barley embryos. In dry embryos, a myelin basic protein (MBP) phosphorylating kinase was detected, which could be immunoprecipitated with an anti-MAPK (mitogen-activated protein kinase) antibody. Rehydration of the embryo induced a decrease in activity of this 45 kDa MAPK-like protein kinase. In addition, activity of a MBP kinase of the same molecular weight was subsequently found to be induced. This second MBP kinase activity could not be immunoprecipitated with the anti-MAPK antibody and was induced only in germinating embryos, not in dormant embryos. PMID- 11056222 TI - Uterine artery embolization for symptomatic uterine myomas. AB - OBJECTIVE: To evaluate the role of uterine artery embolization as treatment for symptomatic uterine myomas. DESIGN: Medline literature review, cross-reference of published data, and review of selected meeting abstracts. RESULT(S): Results from clinical series have shown a consistent short-term reduction in uterine size, subjective improvement in uterine bleeding, and reduced pain following treatment. Posttreatment hospitalization and recovery tend to be shorter after uterine artery embolization compared with hysterectomy. Randomized controlled trials have not been conducted, and long-term efficacy has not been studied. A limited number of deliveries have been reported following uterine artery embolization for uterine myomas. CONCLUSION(S): Uterine artery embolization is a unique new treatment for symptomatic uterine myomas. Even without controlled studies, demand for this procedure has increased rapidly. Uterine artery embolization may be considered an alternative to hysterectomy, or perhaps myomectomy, in well selected cases. At the present time, however, uterine artery embolization should not be routinely recommended for women who desire future fertility. PMID- 11056223 TI - The time has come to simplify the evaluation of the hirsute patient. PMID- 11056224 TI - Human somatic cell nuclear transfer (cloning). The Ethics Committee of the American Society for Reproductive Medicine. PMID- 11056225 TI - Predictors of treatment failure for ectopic pregnancy treated with single-dose methotrexate. AB - OBJECTIVE: To determine variables that predict treatment failure after methotrexate (MTX) treatment of ectopic pregnancy. DESIGN: Retrospective cohort study. SETTING: Canadian teaching hospital. PATIENT(S): Sixty patients diagnosed with and treated for ectopic pregnancy. INTERVENTION(S): A single dose of methotrexate (50 mg/m(2)) by i.m. injection. MAIN OUTCOME MEASURE(S): Resolution of serum beta-hCG or clinical evidence of treatment failure. RESULT(S): Treatment failure was observed following methotrexate administration in 65% of cases when initial beta-hCG was >4000 IU/L, but in only 7. 5% of patients when serum beta hCG was <4000 IU/L (OR = 52.06, 95% CI 4.88-555.56). Patients who presented with pelvic pain without tenderness had treatment failure 56% of the time versus only 17% in those without pain (OR = 9.20, 95% CI 1.02-82.60). Treatment failure also occurred in 53% of patients presenting with vaginal bleeding versus 16% without bleeding (OR = 6.18, 95% CI 0.73-51.93). CONCLUSION(S): Methotrexate should not be used to treat ectopic pregnancy when initial beta-hCG is >4000 IU/L. Caution should also be exercised in using methotrexate for ectopic pregnancy when the patient presents with bleeding or pain even without tenderness. PMID- 11056226 TI - Recent declining trend in ectopic pregnancy in France: evidence of two clinicoepidemiologic entities. AB - OBJECTIVE: To assess the recent incidence of ectopic pregnancy (EP) in France (1992-1997) and to relate this incidence to trends in risk factors and use of contraception. DESIGN: Population register-based study. SETTING: Auvergne EP register (central France). PATIENT(S): Women aged 15-44 years with EP in the Auvergne region. MAIN OUTCOME MEASURE(S): Rates of EP, frequencies of exposure to risk factors, and sales of contraceptive methods in 1992-1997. RESULT(S): The overall EP rate decreased 13.7%, from 96.4 per 100,000 women aged 15-44 in 1992 to 83.2 per 100,000 in 1997. The rate of EP associated with reproductive failure remained stable, but the rate of EP associated with contraceptive failure (mostly intrauterine device failure) decreased 26.6%. The trends in the prevalence of the main risk factors for EP and sales of contraceptive methods are concordant with the changes in EP rates: Risk factor prevalence did not change over time, but intrauterine device sales in the area declined in parallel. CONCLUSION(S): The rates of EP as the result of contraceptive failure and as a result of reproductive failure evolve differently in the population and should not be confused in epidemiologic studies. This finding, along with published evidence that the two types of EP have different risk factors, location, prognosis, and perception by women, indicates that they are two distinct clinical entities possibly requiring different management. PMID- 11056227 TI - Bilateral ectopic pregnancy after transfer of two embryos. AB - OBJECTIVE: To report a case of bilateral tubal ectopic pregnancy (EP) after the transfer of two embryos. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 43-year-old multigravida with bilateral tubal pregnancy. INTERVENTION(S): Operative laparoscopy with right linear salpingostomy and left salpingectomy. MAIN OUTCOME MEASURE(S): Laparoscopy revealed an unruptured left isthmic tubal EP and an unruptured right ampullary tubal EP. RESULT(S): Pathology confirmed immature placental villi in the right tube and placental tissue in the left tube. The patient was discharged home without incident on the day after surgery. CONCLUSION(S): This is a rare case of bilateral tubal pregnancy after the transfer of only two embryos. It is critical to perform a close inspection of the abdomen, pelvis, and contralateral tube after surgery for EP. PMID- 11056228 TI - Endometrial stripe thickness in tubal and intrauterine pregnancies. AB - OBJECTIVE: To evaluate endometrial stripe thickness (EST) among patients with tubal pregnancy (TP) and intrauterine pregnancy (IUP). DESIGN: Historical cohort. SETTING: City hospital. PATIENT(S): Ninety-four women suspected to have TP. INTERVENTION(S): Serum betaHCG and sonographic EST measurements. MAIN OUTCOME MEASURE(S): Comparison of age, gestational age (GA), EST, and log beta HCG. RESULT(S): The two groups of women, 65 with TP and 29 with IUP, had similar mean ages (+/-SD) of 28.6 +/- 5.7 and 28.6 +/- 6.1, respectively. The median values of GA in the 2 groups were similar, 46.6 and 44.6 d, respectively. The mean values for EST (+/-SD), adjusted for GA, were significantly different: 9.9 +/- 5.9 mm in the TP group and 12.6 +/- 5.3 mm in the IUP group. The mean values (+/-SD) of log beta HCG in the 2 groups also differed significantly: 6.90 +/- 1.29 and 7.52 +/- 0.97, respectively. No correlation was found between EST and GA or log beta HCG within either group. CONCLUSION(S): The mean EST in women with TP was significantly smaller than in women with IUP. The wide range of EST values and their overlap precludes the utilization of EST as a single feature in the diagnosis of a tubal pregnancy. PMID- 11056229 TI - Requesting information about and obtaining reversal after tubal sterilization: findings from the U.S. Collaborative Review of Sterilization. AB - OBJECTIVE: To determine the cumulative probabilities over 14 y of requesting information on sterilization reversal and of obtaining a reversal and to identify risk factors observable at sterilization for both measures of regret. DESIGN: The U.S. Collaborative Review of Sterilization, a prospective cohort study. SETTING: Fifteen medical centers in 9 cities. PATIENT(S): 11,232 women. MAIN OUTCOME MEASURE(S): Cumulative probabilities of requesting information on reversal and undergoing reversal. RESULT(S): The 14-y cumulative probability of requesting reversal information was 14.3% (95% confidence interval [CI], 12.4%-16.3%). Among women aged 18 to 24 y at sterilization, the cumulative probability was 40.4% (95% CI, 31.6%-49.2%). Women aged 18 to 24 y were almost 4 times as likely to request reversal information as were women > or = 30 years of age (adjusted rate ratio [RR], 3.5; 95% CI, 2.8-4.4). Number of living children was not associated with requesting reversal information. The overall cumulative probability of obtaining reversal was 1.1% (95% CI, 0.5-1.6). Younger women (18 to 30 y) were more likely to obtain reversal (RR, 7.6; 95% CI, 3.2-18.3). CONCLUSION(S): Women who were sterilized at a young age had a high chance of later requesting information about reversal, regardless of their number of living children. PMID- 11056230 TI - Risk factors for extrauterine pregnancy in women using an intrauterine device. AB - OBJECTIVE: To identify the risk factors for ectopic pregnancy (EP) in women using an intrauterine device (IUD). DESIGN: Case-control study. SETTING: Auvergne region (France). PATIENT(S): Women using an IUD and suffering EP (243 cases) or having an intrauterine pregnancy (140 controls). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Sociodemographic characteristics, smoking, medical history, and medicines taken before the pregnancy. Type of IUD, duration of use, position and visibility of the thread at diagnosis, and presence of abnormal clinical signs. RESULT(S): Seven factors were associated with an increase in the risk of EP: histories of spontaneous abortion, IUD use, and tubal damage; progesterone IUD at the time of conception; insertion of an IUD during the month following a previous pregnancy; duration of use of the IUD in place at the time of conception; and pelvic pain resulting in medical consultation after the insertion of the IUD. Conversely, five factors were associated with a decrease in the risk of EP: history of treated low genital tract infection; history of contraception using the progestagen pill; use of paracetamol or aspirin before the pregnancy; and displacement of the IUD. CONCLUSION(S): This study suggests that the IUD itself may have an etiological role in EP. PMID- 11056231 TI - Y chromosome analysis of infertile men and their sons conceived through intracytoplasmic sperm injection: vertical transmission of deletions and rarity of de novo deletions. AB - OBJECTIVE: To determine the prevalence and type of Yq microdeletions in 86 consecutive men that fathered 99 sons by intracytoplasmic sperm injection (ICSI) and to determine the incidence of vertical transmission and de novo deletions in these boys. DESIGN: Prospective clinical observational study. SETTING: Genetics laboratory associated with a university IVF unit. PATIENT(S): Eighty-six consecutive infertile men presenting to an IVF clinic and their 99 ICSI-conceived sons. Fifty of the 86 men (58%) had idiopathic seminiferous tubule failure (STF); the remainder had a variety of other clinical indications for ICSI. INTERVENTION(S): Collection of peripheral and cord blood samples. MAIN OUTCOME MEASURE(S): The Yq genetic status of fathers who underwent ICSI and of their sons by the presence or absence of 22 Y-specific markers covering the four azoospermia factor (AZF) subregions. RESULT(S): Yq deletions of the AZFd/c region were detected in two (6.9%) of 29 azoo- or severely oligospermic men with STF. Identical deletions were found in their respective sons. No de novo deletions were detected in the remaining 97 sons conceived by men without deletions. CONCLUSION(S): The detection of Yq deletions only in men with severe STF is consistent with previous studies, with the AZFd/c region being most commonly affected. This study demonstrates the vertical transmission of these Yq deletions through the use of ICSI and supports the notion that, in most cases, Yq deletions will be inherited by male offspring. The absence of de novo Yq deletions in the male offspring indicates that these events are rare following ICSI in men with both STF and other common male factor indications. PMID- 11056232 TI - A simple and reliable method for meiotic studies on testicular samples used for intracytoplasmic sperm injection. AB - OBJECTIVE: To develop a reliable and simple method allowing meiotic studies to be performed on testicular samples used for ICSI. DESIGN: Evaluation of meiotic abnormalities in patients with severe spermatogenic impairment. SETTING: Centre de Medecine de la Reproduction, Marseille. PATIENT(S): Two azoospermic men undergoing testicular biopsy for ICSI and one control individual with normal testicular histology. INTERVENTION(S): The immature germ cells from the patients came from testicular biopsy used for ICSI, after dispersal into a thin cell suspension. Cells were cytocentrifuged to obtain well-spread spermatocytes and then immunocytochemical techniques were performed. We used rabbit polyclonal antibodies against the specific meiotic proteins Cor1 and Syn1 and a human CREST anti-kinetochore antibody. MAIN OUTCOME MEASURE(S): Synapsis abnormalities in patients with severe spermatogenesis impairment. RESULT(S): Pachytene spermatocytes are easily analyzed with this technique, without damage of the axial core and synaptonemal complex. The loss of germ cells is limited. CONCLUSION(S): The cytocentrifugation method is the most suitable technique for meiotic studies in patients with severe spermatogenic failure, because it can be used on the testicular cell suspension remaining after ICSI with testicular spermatozoa. PMID- 11056233 TI - Microsurgical single tubular epididymovasostomy: assessment in the era of intracytoplasmic sperm injection. AB - OBJECTIVE: To reevaluate the role of microsurgical single tubular epididymovasostomy for the treatment of obstructive azoospermia in the era of intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective clinical study. SETTING: University infertility clinic. PATIENT(S): Sixty-one patients with obstructive azoospermia who underwent microsurgical single tubular epididymovasostomy. INTERVENTION(S): Microsurgical single tubular epididymovasostomy. MAIN OUTCOME MEASURE(S): The overall patency and live-birth rates and factors that influenced the surgical outcome. RESULT(S): The overall patency rate after surgery was 68.9% (42/61) and the live-birth rate 31.1% (19/61). Of the 19 live-birth cases, 11 were achieved by natural means and 2 were achieved by conventional IVF soon after the operation, then subsequently by natural conception. The remaining 6 were the result of conventional IVF after surgery. An analysis of the potential prognostic factors previously associated with epididymovasostomy indicated that none had a statistically significant correlation with surgical outcome. In cases of patency, the partners were stratified into a younger group (21-30 years; n = 12) and an older group (31-36 years; n = 30). There was no statistically significant difference between the groups in the live-birth rate regardless of the means of conception (natural versus conventional IVF). CONCLUSION(S): This study demonstrates that the results obtained by microsurgical single tubular epididymovasostomy are comparable to those obtained with the use of IVF and ICSI. Even in this era of ICSI, the option of microsurgical single tubular epididymovasostomy should be considered because ICSI involves surgery to retrieve sperm and complex invasive treatment of the wife. PMID- 11056234 TI - Sperm chromosome analysis and outcome of IVF in patients with non-mosaic Klinefelter's syndrome. AB - OBJECTIVE: The aim of the study was to determine the potential risk for fetal chromosomal anomalies in non-mosaic Klinefelter's syndrome patients undergoing IVF and intracytoplasmic sperm injection. DESIGN: Individually collected spermatozoa were isolated from wet testicular tissue preparations and fixed on glass slides using micromanipulation. Their nuclei were analyzed for chromosomes X, Y, and 18 by fluorescent in situ hybridization. SETTING: Assisted reproductive technology program. PATIENT(S): Consenting patients with non-mosaic Klinefelter's syndrome undergoing testicular biopsy and IVF (fresh specimens) or following such treatment (cryopreserved specimens). INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The rates of numerical chromosome abnormalities for chromosomes X, Y, and 18 among spare testicular sperm and the pregnancy outcome following treatment. RESULT(S): Testicular sperm were found in 8 of 20 patients. Four couples became pregnant following embryo replacement. Sperm chromosomes were analyzed in five patients. One hundred and five sperm of 112 analyzed (93.7%) were normal with X to Y ratio of 50:55 (NS) respectively. Among the 112 sperm tested, seven (6.3%) demonstrated chromosomal abnormalities, of which five were related to the sex chromosomes and two to chromosome 18. One set of triplets, one set of twins, and two singletons (four males and three females) with normal karyotypes were born. CONCLUSION(S): Most of the testicular sperm retrieved from Klinefelter's syndrome patients demonstrates a normal pattern of sex chromosome segregation. Therefore, the risk of transmitting numerical sex chromosome abnormalities is relatively low and probably comparable with the rates found in other severe male factor infertility patient groups. PMID- 11056235 TI - Cigarette smoking and the risk of male factor subfertility: minor association between cotinine in seminal plasma and semen morphology. AB - OBJECTIVE: To evaluate the impact of cigarette smoking on male factor subfertility and the semen parameters of sperm count, motility, and morphology by questionnaire and determination of the cotinine concentrations in blood and seminal plasma of fertile and subfertile males. DESIGN: Case-control study of 107 fertile and 103 subfertile males who provided a standardized blood and semen specimen and completed a self-administered questionnaire about their smoking habits. SETTING: Outpatient fertility clinic of the University Medical Centre St. Radboud, Nijmegen, The Netherlands. PATIENT(S): One hundred seven fertile and 103 subfertile males. INTERVENTION(S): Vena puncture and semen collection. MAIN OUTCOME MEASURE(S): Blood and seminal plasma cotinine levels in relation to semen parameters. RESULT(S): A higher frequency of cigarette smoking was observed in subfertile males than in fertile males, with an odds ratio of 1.7 (95% confidence interval, 0.9-3.2). The self-reported number of cigarettes smoked per day correlated with the cotinine concentrations in blood and seminal plasma for both groups. A small but statistically significant correlation was found between cotinine concentrations in seminal plasma and the percentage of abnormal sperm morphology, but not for other semen parameters (r(s) = 0.19). CONCLUSION(S): Although the mechanism of the toxicity of cotinine on sperm morphology is not clear, this study indicates only a minor effect of cigarette smoking on male factor subfertility, which is probably due to compounds in cigarette smoke other than nicotine (cotinine). PMID- 11056236 TI - Double (consecutive) transfer of early embryos and blastocysts: aims and results. AB - OBJECTIVE: The aim of this study was to evaluate the consecutive transfer approach of early embryos and blastocyst(s). DESIGN: Case-control study. SETTING: Public assisted reproduction technology unit. PATIENT(S): The study population consisted of three groups. In Group 1, a double transfer was performed on 136 consecutive women, that is, a standard transfer of embryos on day 2 or 3, and a second transfer of a blastocyst(s). In Group 2, an early transfer of only two embryos and a second transfer of one blastocyst were performed on 29 women from group 1 who had more than three high-quality embryos available for early transfer. In Group 3, a single early transfer was performed on 139 consecutive women who received three high-quality embryos (controls). INTERVENTION(S): Early embryo transfer, extended culture of the spare embryos, and a second transfer of a blastocyst(s). MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates. RESULTSs): No differences were detected among the three groups in either pregnancy or implantation rates (pregnancy: 36.8%, 41.4%, and 37.4%, respectively; implantation: 14.6%, 19.9%, and 19.8%, respectively). CONCLUSION(S): The double (consecutive) transfer of early embryos and blastocyst(s) does not offer any advantage over the traditional early transfer. This may be from the adverse effect of the second transfer on the implantation process. PMID- 11056237 TI - Effects of functional ovarian cysts detected on the 7th day of gonadotropin releasing hormone analog administration on the outcome of IVF treatment. AB - OBJECTIVE: To investigate the impact of functional ovarian cysts on the time required to achieve pituitary suppression, follicular development, embryo quality, and pregnancy rates during IVF treatment. DESIGN: Prospective observational study. INTERVENTION(S): Daily treatment with buserelin (sc 500 microg) was initiated on day 2 of menstruation. Ultrasound and hormonal tests were performed on days 1, 7, 11, 14, and weekly thereafter until pituitary suppression was achieved. RESULT(S): 48 patients underwent 51 cycles of IVF treatment. A functional cyst was detected in three cycles (5.8%) with baseline ultrasound scan and in 27 cycles (52.9%) on day 7 of buserelin administration. Patients who developed a cyst required a significantly longer time to achieve pituitary suppression (21 vs. 7 days), had a significantly lower FSH level at the time of initiation of gonadotropins, required more ampules of gonadotropin (45 vs. 41 ampules), developed less follicles (13 vs. 17.5), and had lower embryo quality. However, there were no differences in the implantation (23.5% vs. 17.2%) and pregnancy rates (37.2% vs. 29.2%) between two groups. CONCLUSION(S): Functional cysts prolong the period to achieving pituitary suppression, increase gonadotropin requirements, and decrease follicular recruitment and embryo quality. They have, however, no negative effect on pregnancy rates. PMID- 11056238 TI - Prospective randomized study of two cryopreservation policies avoiding embryo selection: the pronucleate stage leads to a higher cumulative delivery rate than the early cleavage stage. AB - OBJECTIVE: To compare the cumulative live birth rates obtained after cryopreservation of either pronucleate (PN) zygotes or early-cleavage (EC) embryos. DESIGN: Prospective randomized study. SETTING: University hospital. PATIENT(S): Three hundred eighty-two patients, involved in an IVF/ICSI program from January 1993 to December 1995, who had their supernumerary embryos cryopreserved either at the PN (group I) or EC (group II) stage. For 89 patients, cryopreservation of EC embryos was canceled because of poor embryo development (group III). Frozen-thawed embryo transfers performed up to December 1998 were considered. MAIN OUTCOME MEASURE(S): Age, oocytes, zygotes, cryopreserved and transferred embryos, damage after thawing, cumulative embryo scores, implantation, and cumulative live birth rates. RESULT(S): The clinical pregnancy and live birth rates were similar in all groups after fresh embryo transfers. Significantly higher implantation (10.5% vs. 5.9%) and pregnancy rates (19.5% vs. 10.9%; P< or = .02 per transfer after cryopreserved embryo transfers were obtained in group I versus group II, leading to higher cumulative pregnancy (55.5% vs. 38.6%; P < or = .002 and live birth rates (46.9% vs. 27.7%; P< or = .0001. CONCLUSION(S): The transfer of a maximum of three unselected embryos and freezing of all supernumerary PN zygotes can be safely done with significantly higher cumulative pregnancy chances than cryopreserving at a later EC stage. PMID- 11056239 TI - Follicular fluid concentrations of interleukin-12 and interleukin-8 in IVF cycles. AB - OBJECTIVE: To investigate the role of interleukin-12 (IL-12) and IL-8 in the periovulatory follicular fluid during in vitro fertilization cycles. DESIGN: A prospective study. SETTING: Reproductive Medicine Unit, Liverpool Women's Hospital, United Kingdom. PATIENT(S): Women undergoing in vitro fertilization treatment. INTERVENTION(S): IL-8 and IL-12 concentrations in follicular fluid samples that had been collected during transvaginal oocyte retrieval were measured using an enzyme-linked immunosorbent assay (ELISA). Cytokine concentrations were correlated to fertilization rates and treatment outcome. MAIN OUTCOME MEASURE(S): Fertilization rates and ultrasonographic evidence of intrauterine pregnancy by 4 weeks after embryo transfer. RESULT(S): Failed fertilization in women with detectable IL-12 was significantly higher (45.5%) than in the IL-12 negative group (6.1%), P=.01. None of the women with detectable IL-12 achieved a pregnancy at the end of the treatment (P=.01). IL-8 was present in the follicular fluid of all women, and no difference in its concentrations was found between the pregnant and nonpregnant groups. No correlation was found between the follicular fluid concentrations of IL-8 and fertilization rates. CONCLUSION(S): The presence of IL-12 in the follicular fluid appears to be associated with a negative outcome in IVF treatment. Interleukin-8 appears to be an essential part of folliculogenesis, although its concentration is not associated with fertilization or implantation rates. PMID- 11056240 TI - Luteinizing hormone depletes ascorbic acid in preovulatory follicles. AB - OBJECTIVE: The objective of the present studies was to determine whether luteinizing hormone (LH) depletes ascorbic acid in the preovulatory follicle. DESIGN: Controlled, prospective experimental study. SETTING: University-based research center. ANIMAL(S): Sprague-Dawley female rats. INTERVENTION(S): Follicular growth and ovulation were induced in immature rats by gonadotropin treatment. MAIN OUTCOME MEASURE(S): Analysis of ovary, follicle, and oocyte levels of ascorbic acid by colorimetric analysis and high-pressure liquid chromatography. RESULT(S): Ovarian ascorbic acid was maximally depleted (50%) within 2 h of LH treatment and was sustained for 8 h. Follicle ascorbic acid levels were unchanged 1 h after LH injection but were significantly reduced within 2 h (40%). Incubation of isolated preovulatory follicles for 3 h with hCG or with menadione (a generator of oxygen radicals) reduced ascorbic acid levels. Isolation of cumulus-enclosed or denuded oocytes depleted ascorbic acid to undetectable levels, but follicular ascorbic acid levels were only moderately depleted by isolation and incubation. Accumulation of ascorbic acid by oocytes was significantly enhanced by the presence of intact cumulus cells. CONCLUSION(S): Elevation of LH and the production of oxygen radicals deplete ascorbic acid in the preovulatory follicle. PMID- 11056241 TI - Long-term use of gonadotropin-releasing hormone analogs and hormone replacement therapy in the management of endometriosis: a randomized trial with a 6-year follow-up. AB - OBJECTIVE: To identify the effects of long-term GnRH agonist use (6-24 months), with and without add-back therapy, and spontaneous reversibility of bone mass density (BMD) up to 6 years after treatment. DESIGN: A prospective, randomized, long-term follow-up study. SETTING: Obstetrics and gynecology department in a university hospital in the United Kingdom. PATIENT(S): Forty-nine symptomatic women with a laparoscopic diagnosis of endometriosis who had been identified for treatment with long-acting GnRH agonist and volunteered to participate in the study. INTERVENTION(S): Women were randomly allocated to receive hormone replacement therapy (HRT) as a daily oral dose of estradiol, 2 mg, and norethisterone acetate, 1 mg, or no treatment in addition to monthly subcutaneous implants of goserelin acetate for up to 2 years, until cessation of symptoms. Bone mineral density (BMD) at the lumbar spine (C2-C4) and hip (Ward triangle) was measured every 6 months. MAIN OUTCOME MEASURE(S): BMD changes in both groups. RESULT(S): 45 women were followed up for 6 years, at the end of which the groups did not differ significantly in the reduction in mean BMD at the lumbar spine or hip. CONCLUSION(S): BMD reduction occurs during long-term GnRH agonist use and is not fully recovered by up to 6 years after treatment. Use of HRT does not affect this process. PMID- 11056242 TI - Effect of hormonal agents on monocyte chemotactic protein-1 expression by endometrial epithelial cells of women with endometriosis. AB - OBJECTIVE: To assess whether hormonal agents used in the medical treatment of endometriosis, such as danazol and GnRH agonist, exert direct regulatory action on monocyte chemotactic protein-1 (MCP-1) expression by endometrial epithelial cells. DESIGN: Primary cultures of epithelial cells isolated from human endometrium were exposed to different concentrations of cytokines and steroid hormone analogs. Expression of MCP-1 was analyzed at the levels of protein and messenger RNA. SETTING: Gynecology clinic and laboratory of endocrinology of reproduction. PATIENT(S): Women presenting for infertility or pelvic pain in whom endometriosis was diagnosed by using laparoscopy. INTERVENTION(S): Endometrial tissue biopsy performed at laparoscopy. MAIN OUTCOME MEASURE(S): Secretion of MCP 1 protein was measured by using enzyme-linked immunosorbent assay, and mRNA steady-state levels were measured by performing Northern blot analysis. RESULT(S): Buserelin acetate, a GnRH agonist (0.1-10 ng/mL), had no significant effect on MCP-1 expression, whereas danazol (10(-7)-10(-5) M), a testosterone analog, and dexamethasone, an anti-inflammatory glucocorticoid hormone (10(-12) 10(-6)M), showed a direct and a dose-dependent inhibitory effect on MCP-1 expression. This effect occurred at the level of protein and mRNA. CONCLUSION(S): The findings of the study may affect understanding of the mechanisms by which hormonal treatments act on endometriosis and influence its clinical manifestations. PMID- 11056243 TI - Mutation analysis of the growth differentiation factor-9 and -9B genes in patients with premature ovarian failure and polycystic ovary syndrome. AB - OBJECTIVES: To investigate whether growth differentiation factor (GDF)-9 or GDF 9B/bone morphogenetic protein (BMP)-15 mutation is present in Japanese women with premature ovarian failure (POF) and polycystic ovary syndrome (PCOS). DESIGN: Clinical and molecular studies. SETTING: Outpatient clinic in a university hospital. PATIENT(S): Fifteen women with POF, 38 women with PCOS, and 3 normal fertile controls. INTERVENTION(S): Extraction of DNA from blood samples for subsequent polymerase chain reaction (PCR). MAIN OUTCOME MEASURE(S): Amplified DNA was analyzed by denaturing gradient gel electrophoresis (DGGE) and/or by direct sequencing. RESULT(S): No missense mutation was found in any exons of the GDF-9 gene and the GDF-9B/BMP-15 gene in patients with POF and PCOS. CONCLUSION(S): The missense mutation in the GDF-9 gene or the GDF-9B/BMP-15 gene is uncommon in anovulatory Japanese women with POF and PCOS. PMID- 11056244 TI - Examination of the chin or lower abdomen only for the prediction of hirsutism. AB - OBJECTIVE: To test the hypothesis that scoring terminal hair growth on only the chin or abdomen can serve as a reliable predictor for hirsutism. DESIGN: A prospective observational study. PATIENT(S): Six hundred and ninety-five consecutive hyperandrogenic women seen between June 1987 and December 1997. MAIN OUTCOME MEASURE(S): All hirsutism exams were performed by one examiner. Hirsutism was scored using a modification of the Ferriman-Gallwey (F-G) method. An F-G score of > or = 8 defined hirsutism. RESULT(S): Of the 695 women examined 352 (50.1%) had hirsutism scores of 8. Thirty percent (79 of 344) of women who had an F-G score of <8 had previously underwent electrology. If either the chin or lower abdomen hair growth score was > or = 2, the sensitivity was 100% for the prediction of hirsutism, although the specificity was 27%. The positive predictive value (PPV) for hirsutism using a hair score of > or = 2 at either of these sites was 58%. CONCLUSION(S): A hair growth score of > or = 2 on the chin or lower abdomen only was found to be a highly sensitive predictor for hirsutism. However, because of its very low PPV, this screening method is virtually useless in populations where the hirsutism frequency is expected to be low, about 5%. However, this screening method for the detection of hirsutism would be useful for the study of high-risk populations with an expected hirsutism prevalence of >20% (e.g., family studies). PMID- 11056245 TI - Depressive mood symptoms associated with ovarian suppression. AB - OBJECTIVE: To determine if sertraline is helpful in the management of depressive symptoms associated with ovarian suppression during GnRH agonist therapy as compared with a placebo-controlled group. DESIGN: Double-blind placebo-controlled prospective study design. SETTING: An obstetrics/gynecological office specializing in infertility in an academic environment. PATIENT(S): Premenstrual women with laparoscopically diagnosed endometriosis who required GnRH agonist therapy for treatment and did not have significant depressive or premenstrual mood symptoms at baseline. INTERVENTION(S): Participants were randomly assigned to either the sertraline treatment group or to the placebo group for the 3-month duration of the GnRH agonist therapy. MAIN OUTCOME MEASURE(S): The 21-item Hamilton Rating Scale for Depression (HRSD), which is an instrument designed to assess depressive symptomatology. RESULT(S): A Hotellings T(2) test for repeated measure analysis indicated a statistically significant (P<.05) between-group difference across time for the HRSD (T(2) = 13.3; F[3, 28] = 4.1; P=.02) with the sertraline treatment group manifesting significantly fewer depressive symptoms than the control group. CONCLUSION(S): The results indicate that sertraline is an effective option in the management of depressive mood symptoms associated with ovarian suppression during GnRH agonist therapy. PMID- 11056246 TI - Primary habitual abortions are associated with high frequency of factor V Leiden mutation. AB - OBJECTIVE: To analyze the prevalence of the mutation G1691A in factor V gene (Leiden mutation), of mutation C677T in the methylenetetrahydrofolate reductase (MTHFR) gene, and of polymorphism in G20210A in the prothrombin gene in women with recurrent abortions; further, to identify a subgroup at higher risk of being carriers of these mutations. DESIGN: Prospective case control evaluation. SETTING: University clinic. PATIENT(S): Eighty-four women with 3 or more consecutive miscarriages were compared with 69 controls. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Polymerase chain reactions were performed to identify the mutations G1691A in factor V and C677T in MTHFR genes and the polymorphism G20210A in the prothrombin gene. RESULT(S): In women with primary habitual abortions, 27.8% carried the Leiden mutation. No difference was observed in the prevalence of mutation C677T in the MTHFR gene or in polymorphism G20210A in the prothrombin gene. CONCLUSION(S): The Leiden mutation may play a considerable role for women having primary recurrent abortions. PMID- 11056247 TI - No evidence of somatic activating mutations on gonadotropin receptor genes in sex cord stromal tumors. AB - OBJECTIVE: To search for somatic activating mutations of gonadotropin receptor (FSH-R and LH/chorionic gonadotropin receptor [CG-R]) genes as a cause of sex cord stromal tumors. DESIGN: Molecular studies in human tissue. SETTING: University hospital. SPECIMEN(S): Eight granulosa cell tumors collected from paraffin-embedded tissue, eight Leydig cell tumors, and three thecomas collected from fresh-frozen or paraffin-embedded tissue. INTERVENTION(S): Tumor samples were used for DNA extraction. The entire exon 11 of the LH/CG-R gene and a hot spot for gonadotropin receptor activating mutations on exon 10 of the FSH-R gene were amplified by polymerase chain reaction. The former was analyzed by denaturing gradient gel electrophoresis and automatic direct sequencing, and the latter by automatic direct sequencing. MAIN OUTCOME MEASURE(S): Results of denaturing gradient gel electrophoresis and automatic direct sequencing. RESULT(S): No somatic activating mutation was detected in exon 11 of the LH/CG-R gene in eight Leydig cell tumors and three thecomas. In addition, no mutations were detected in eight granulosa cell tumors in the hot spot for activating mutations in exon 10 of the FSH-R gene. CONCLUSION(S): Somatic activating mutations of gonadotropin receptors seem to play no relevant role in the development of sex cord stromal tumors. PMID- 11056248 TI - Embryotropic role of hemoglobin and ethylenediaminetetraacetic acid in preimplantation development of ICR mouse 1-cell embryos. AB - OBJECTIVE: To evaluate embryotropic action of hemoglobin (Hb) and ethylenediaminetetraacetic acid (EDTA) on preimplantation embryo development. DESIGN: In vitro model study using mouse embryos. SETTING: University affiliated hospital, Pochon CHA University. ANIMALS: Four-week-old block strain ICR mice naturally mated after superovulation. INTERVENTION(S): One-cell embryos were cultured in serum-free, modified preimplantation-1 medium, to which 1 microg/ml Hb and/or 0.1 mM EDTA were added. MAIN OUTCOME MEASURE(S): Preimplantation development and blastomere number. RESULT(S): More (P<.05) 1-cell embryos developed to the 4-cell (52% vs. 67%-84%), 8-cell (48% vs. 65%-81%), and blastocyst (40% vs. 61%-79%) stages after the addition of hemoglobin (Hb) and/or EDTA than after no addition. Highest proportion of embryos developed to each stage after the combined addition of Hb+EDTA. EDTA specifically stimulated the development before the 8-cell stage, which was as similar as Hb+EDTA. On the contrary, higher ratio of morula to blastocyst transformation was obtained after the addition of Hb or Hb+EDTA than after no addition (0.76 vs. 0.96-0.98). Significant increases in the cell number of blastocysts (46.5-47.2 vs. 53.2 cells), inner cell mass (ICM) cells (16.7-17.5 vs. 21 cells), and the ratio of ICM cells to trophoblasts (0.3-0.37 to 0.39) were found after the combined addition of Hb+EDTA, compared with no addition or with the addition of EDTA or Hb alone. CONCLUSIONS: Hb and EDTA have stage-specific effects on supporting preimplantation embryo development; Hb promotes both the development before the 8 cell stage and the morula to blastocyst transformation, whereas EDTA mainly promotes the development to the 8-cell stage. The combined exposure of embryos to Hb and EDTA improves not only preimplantation development but also the growth and quality of blastocysts. PMID- 11056249 TI - Effect of the gonadotropin-releasing hormone antagonist ganirelix on cyclic adenosine monophosphate accumulation of human granulosa-lutein cells. AB - OBJECTIVE: To evaluate whether the GnRH antagonist ganirelix exerts an effect on cyclic adenosine monophosphate (cAMP) production of human granulosa-lutein (GL) cells in vitro. DESIGN: In vitro cell culture study. SETTING: Research laboratory of a university hospital. PATIENT(S): Mural GL and cumulus cells were obtained from 15 patients on whom controlled ovarian hyperstimulation was being performed for intracytoplasmic sperm injection treatment. INTERVENTION(S): Mural GL and cumulus cells were cultured for 48 hours with and without 1 nM ganirelix or triptorelin. For the last 6 hours, the cells were either exposed to 1-5 IU hCG or left unstimulated. MAIN OUTCOME MEASURE(S): At the end of the culturing period, the intracellular and extracellular cAMP accumulations were measured by an (125)I scintillation proximity assay. RESULT(S): hCG induced dose-dependent increases in total cAMP accumulation. Stimulation with 1 IU/mL hCG resulted in 9-fold and 13 fold increases, and 5 IU/mL hCG resulted in 19-fold and 14-fold increases in total cAMP release from cumulus and mural GL cells, respectively. On the other hand, treatments with 1 nM GnRH antagonist ganirelix and 1 nM GnRH agonist triptorelin did not exert any significant changes on the basal and hCG-stimulated cAMP accumulation of mural GL cells and cumulus cells as compared with controls. CONCLUSION(S): Ganirelix does not influence basal and hCG-stimulated cAMP accumulation of human GL cells in vitro. cAMP is apparently not involved in the mechanism of action of GnRH analogs in human ovary. PMID- 11056250 TI - The immune response during the luteal phase of the ovarian cycle: a Th2-type response? AB - OBJECTIVE: To test the hypothesis that during the luteal phase of the ovarian cycle, as compared with the follicular phase, the peripheral immune response is shifted toward a type-2 response. DESIGN: Prospective study. SETTING: Academic research setting. PATIENT(S): Women with regular menstrual cycles. INTERVENTION(S): Blood samples were collected between days 6 and 9 of the menstrual cycle and 6-9 days after the LH surge. MAIN OUTCOME MEASURE(S): Intracellular cytokine production of interferon (IFN)-gamma, interleukin (IL)-2, IL-4, and IL-10 after in vitro stimulation of lymphocytes as well as total white blood cell (WBC) count, differential WBC count, and plasma 17 beta-E(2) and P concentrations. RESULT(S): Mean plasma 17 beta-E(2) and P concentrations, WBC count, and mean granulocyte, monocyte, and lymphocyte counts were significantly increased in the luteal phase as compared with the follicular phase of the ovarian cycle. Production of type-1 cytokines (IFN-gamma and IL-2) and production of the type-2 cytokine IL-10 did not vary between the phases of the ovarian cycle. Production of the type-2 cytokine IL-4, however, was significantly increased in the luteal phase as compared with the follicular phase of the ovarian cycle. CONCLUSION(S): During the luteal phase of the ovarian cycle, the immune response is shifted toward a Th2-type response, as reflected by increased IL-4 production in this phase of the cycle. These results may suggest that increased levels of P and 17 beta-E(2) in the luteal phase of the ovarian cycle play a role in the deviation of the immune response toward a type-2 response. PMID- 11056251 TI - RU486-induced growth inhibition of human endometrial cells. AB - OBJECTIVE: To determine the direct action of RU486 on endometrial cell proliferation and to differentiate whether the antioxidant or the antiprogesterone property of RU486 is predominately responsible for its effect on cell growth. DESIGN: In vitro study comparing the effects of RU486 (antiprogesterone and antioxidant), reduced RU486 (antioxidant), ZK112,993 (antiprogesterone), and lazaroid U74,500A (antioxidant) on endometrial cell growth. The human endometrial cell line EM42 was used in transient transfection assays to confirm the relative antiprogesterone potency of the various compounds. SETTING: Academic medical center PATIENT(S): Women presenting with pelvic pain or infertility and diagnosed with endometriosis at time of surgery or women desiring tubal ligation with a normal pelvis (controls). INTERVENTION(S): Endometrial cell cultures were treated with RU486, reduced RU486, lazaroid U74,500A, and ZK112,993. MAIN OUTCOME MEASURE(S): Tritiated thymidine incorporation was used to assess cell growth. Inhibition of progesterone induction of transiently transfected reporter plasmids was used to measure antiprogesterone activity of compounds studied. RESULT(S): RU486 reduced cell growth in a dose-dependent fashion of the endometrial cell lines EM42 and RL95-2 and of endometrial and endometriosis cells from primary culture. After being reduced, RU486 lost most of its antiprogesterone activity but retained its antiproliferative properties. ZK112,993 was similar in potency to RU486 as a progesterone antagonist but did not significantly modify endometrial cell growth. Lazaroid U74,500A was devoid of antiprogesterone activity but was shown to be a potent antiproliferative agent. CONCLUSION(S): RU486 has a direct inhibitory effect on human endometrial cell growth. This activity appears to be at least partly mediated through its antioxidant property. PMID- 11056252 TI - Robotically assisted laparoscopic microsurgical tubal reanastomosis: a feasibility study. AB - OBJECTIVE: To assess the feasibility and reproducibility of laparoscopic microsurgical tubal anastomosis using a remote-controlled robot. DESIGN: Descriptive case study. SETTING: Academic medical center. PATIENT(S): Eight patients with previous laparoscopic tubal sterilization who requested tubal reanastomosis. INTERVENTION(S): Systematization of the operative steps for laparoscopic tubal reanastomosis using a remote-controlled robot. MAIN OUTCOME MEASURE(S): Primary outcome measures were feasibility and reproducibility; secondary measures were tubal patency, operative time, complications, and ergonomic qualities. RESULT(S): The 16 tubes were successfully reanastomosed and patency was confirmed. The mean time that the robotic system was in use was 140 minutes, and mean surgical time was 52 minutes per tube. CONCLUSION(S): Laparoscopic microsurgical tubal reanastomosis after tubal sterilization can be performed using a remote-controlled robotic system. The robot, which has three dimensional vision, allows the surgeon to perform ultraprecise manipulations with intraabdominal articulated instruments while providing the necessary degrees of freedom. Systematization of the operative steps allowed performance of the operation at a speed that compares favorably with the time needed for open microsurgical techniques. Larger series are needed to assess postoperative pregnancy rates. PMID- 11056253 TI - Ovarian remnant syndrome after laparoscopic oophorectomy. AB - OBJECTIVE: To report the surgical history, clinical characteristics, and operative technique used in patients with ovarian remnant syndrome after laparoscopic oophorectomy. DESIGN: Observational study. SETTING: Teaching hospital and private practice office. PATIENT(S): Nineteen patients with documented history of unilateral or bilateral laparoscopic oophorectomies with histologic confirmation of ovarian remnants. INTERVENTION(S): Operative laparoscopy for resection of ovarian remnants. MAIN OUTCOME MEASURE(S): Risk factors and surgical technique contributing to ovarian remnant syndrome. RESULT(S): The patients underwent a mean of 4.7 previous surgical procedures (range, two to nine): 12 had bilateral oophorectomy, and seven had unilateral oophorectomy. The infundibulopelvic ligament had been secured with bipolar desiccation in 11 patients, pretied surgical loops in six, and a linear stapler in two. Cystic ovarian remnants were identified by pelvic sonography in 12 women and by computed tomography (CT) scan in one. Six women underwent reoperation, two for ovarian remnants in different sites. CONCLUSION(S): With laparoscopic oophorectomy there is risk of ovarian remnant due to improper tissue extraction or misapplication or improper use of pretied surgical loops, linear stapler, or bipolar electrodesiccation on the infundibulopelvic ligament, especially in women with a history of multiple pelvic surgeries, adhesions, or endometriosis. PMID- 11056254 TI - Evaluation of outpatient hysteroscopy, saline infusion hysterosonography, and hysterosalpingography in infertile women: a prospective, randomized study. AB - OBJECTIVE: To compare the diagnostic accuracy, pain scores, and procedure length of outpatient hysteroscopy (OHS), hysterosalpingography (HSG), and saline infusion hysterosonography (SIS) for evaluation of the uterine cavity of infertile women. DESIGN: Prospective, randomized, investigator-blind study. SETTING: Tertiary infertility clinic. PATIENT(S): Forty-six consecutive infertile women. INTERVENTION(S): Outpatient HSG, OHS, and SIS, followed by operative hysteroscopy (HS). MAIN OUTCOME MEASURE(S): Uterine abnormalities, procedure length, and subjective pain. RESULT(S): Fifty-nine percent of infertile subjects were found to have an abnormality on at least one of three outpatient uterine evaluations. When compared with the case of definitive operative HS, 60% of abnormalities were correctly classified by HSG, 72% by OHS, and 52% by SIS (P: NS). When comparing all combinations of 2 outpatient screening tests to operative hysteroscopy, 68% were correctly classified by HSG/OHS, 58% by HSG/SIS, and 64% by OHS/SIS (P: NS). The average time length for the OHS was 9.1 min., which was significantly greater than for both HSG (average, 5.3 min) and SIS (average, 6.1 min.) (P<.0001 for both). HSG and SIS were not statistically different regarding procedure time length. The average pain score (0-10) for SIS was 2.7, compared with 5.8 and 5.3 for HSG and OHS, respectively. Both HSG and OHS mean pain scores were significantly greater than the SIS mean. CONCLUSION(S): OHS, SIS, and HSG were statistically equivalent regarding evaluation of uterine cavity pathology in infertile women. PMID- 11056255 TI - One-step hysteroscopic removal of sinking submucous myoma in two infertile patients. AB - OBJECTIVE: To report one-step resectoscopic removal of submucous myomas that were pushed back into the muscular layer by increased intrauterine pressure during hysteroscopic procedures. DESIGN: Case report. SETTING: Kawasaki Municipal Hospital, Kawasaki, Japan. PATIENT(S): Two infertile women presenting with menorrhagia in whom submucous myoma with a broad base was diagnosed. INTERVENTION(S): One patient was pretreated with GnRH agonist for 4 months; the other patient did not receive this treatment. Resectoscopic myomectomies were performed under close sonographic monitoring. MAIN OUTCOME MEASURE(S): Clinical symptoms and conception status. RESULT(S): Tumor sinking occurred during the hysteroscopic procedures, but complete resectoscopic removal of the submucous myomas was achieved under sonographic and hysteroscopic visualization. One patient experienced hyponatremia but recovered after conservative treatment. Both patients conceived after myoma removal. CONCLUSION(S): Sinking myomas, which may cause infertility, can be removed with a one-step hysteroscopic procedure. Sinking of submucous myomas during hysteroscopy might be caused by pretreatment with GnRH agonist and by increased intrauterine pressure during hysteroscopy. We recommend that intrauterine pressure be <45 mmHg, equivalent to hanging a bag of fluid under gravity control 70 cm above the patient's uterus, at the beginning of operations for sinking myomas. PMID- 11056256 TI - Metrorrhagia caused by chronic hematosalpinges. PMID- 11056257 TI - Bilateral undescended ovaries: association with infertility and treatment with IVF. AB - OBJECTIVE: To describe the clinical findings in a patient with bilateral undescended ovaries and infertility who was successfully treated by IVF. DESIGN: Case report. SETTING: University hospital. PATIENT(S): A 35-year-old woman with bilateral undescended ovaries. INTERVENTION(S): Hysterosalpingography, laparoscopy, and an IVF cycle. MAIN OUTCOME MEASURE(S): Anatomic description and pregnancy test. RESULT(S): This patient conceived and delivered a twin gestation after laparoscopic retrieval of oocytes and transfer of two blastocysts. CONCLUSION(S): Bilateral undescended ovaries is a rare condition that can be associated with infertility but can be successfully treated by IVF. PMID- 11056258 TI - Birth of healthy twins after fertilization with in vitro cultured spermatids from a patient with massive in vivo apoptosis of postmeiotic germ cells. AB - OBJECTIVE: To examine whether the results of assisted reproduction with the use of elongated spermatids from a man with incomplete arrest of spermiogenesis and a high frequency of apoptosis among postmeiotic germ cells can be improved by germ cell in vitro culture. DESIGN: Case report. SETTING: Private assisted reproduction centers and a university department. PATIENT(S): Man with incomplete spermiogenesis failure. INTERVENTION(S): Testicular spermatid extraction, in vitro culture of testicular biopsy samples, intraoocyte injection of elongated spermatids, embryo culture, and transfer. MAIN OUTCOME MEASURE(S): Fertilization rate, embryo morphology, and pregnancy. RESULT(S): An assisted reproduction attempt with viability-selected elongated spermatids from a fresh testicular biopsy sample resulted in a poor fertilization result and embryo quality. Further analysis of the sample used in this attempt showed a high incidence of apoptosis among postmeiotic germ cells. A second attempt was then performed with in vitro culture of testicular cells for 24 hours before spermatid injection. This procedure led to a significant decrease in the frequency of apoptosis among viability-selected spermatids, to improvement of the fertilization rate and embryo quality, and to the birth of healthy twins. CONCLUSION(S): In vitro culture of testicular biopsy samples before assisted reproduction with elongated spermatids may improve the efficacy of treatment in cases of massive in vivo apoptosis of postmeiotic germ cells. PMID- 11056259 TI - Atypical hatching of a human blastocyst leading to monozygotic twinning: a case report. AB - OBJECTIVE: To report a case of monozygotic twinning after atypical hatching of a human blastocyst. DESIGN: Case report. SETTING: University-based IVF program. PATIENT(S): A 33-year-old woman with tubal sterility. INTERVENTION(S): Embryo transfer of a human blastocyst with atypical hatching. MAIN OUTCOME MEASURE(S): Development to the blastocyst stage, hatching process, follow-up of pregnancy. RESULT(S): Development of a monozygotic dichorial pregnancy. CONCLUSION(S): First report of an atypical hatching of a human blastocyst leading to dichorial monozygotic twinning. PMID- 11056260 TI - Genetic amniocentesis in multifetal pregnancies reduced to twins compared with nonreduced twin gestations. PMID- 11056261 TI - Delayed assessment of serum and whole blood estradiol, progesterone, follicle stimulating hormone, and luteinizing hormone kept at room temperature or refrigerated. PMID- 11056262 TI - Effect of androgenic anabolic steroids on semen parameters and hormone levels in bodybuilders. PMID- 11056263 TI - Oocytes for late starters and posterity: are we on to something good or bad? PMID- 11056264 TI - Food and chemical toxicology. PMID- 11056265 TI - Dragon's Blood incense: misbranded as a drug of abuse? AB - An unknown red substance was being sold and used with other drugs of abuse in Virginia (often being used in conjunction with marihuana). The red substance was identified as Dragon's Blood incense from Daemonorops draco. In bioassays, Dragon's Blood incense exhibited a low, but measurable cytotoxicity in in vitro cell lines. Dragon's Blood incense or Volatilized Dragon's Blood had no adverse effect on mouse motor performance based on the inclined screen and rotorod tests. delta(9)-Tetrahydrocannibinol (THC) produced a dose-related decline in mouse performance on the rotorod test. The combination of Dragon's Blood incense or Volatilized Dragon's Blood with delta(9)-THC did not contribute further to the impairment of the mice on the rotorod. This data suggests that the abuse potential for Dragon's Blood incense alone or in combination with marihuana is minimal. PMID- 11056266 TI - Is the appearance of macrophages in pulmonary tissue related to time of asphyxia? AB - In order to connect the appearance of macrophages and giant cells in pulmonary tissue with the time of asphyxia the authors analyzed 50 asphyxiated human lungs paying their attention on the number of alveolar and interstitial macrophages and giant cells. They compared histological specimens of 25 asphixiated humans lungs following a slow asphyxia (30 min or more) with 25 histological specimens of asphyxiated human lungs following a rapid asphyxia (10-15 min). Alveolar and interstitial macrophages and giant cells per section, were considered and numbered. Controls were done on histological examination of traumatized lungs. In the pulmonary alveoli following on acute asphyxia there were 27.7+/-4.4 macrophages per section. Subjects dead after a slow asphyxiation showed 68.2+/ 7.1 alveolar macrophages per section (p<0.001). Interstitial macrophages were also frequently present. No differences are detectable in the number of polynuclear giant cells between rapidly and slowly asphyxiated human lungs. The number of alveolar and interstitial macrophages per section can be considered as a further histological evidence of a slow asphyxia and can differentiate a slow asphyxia from an acute one. PMID- 11056267 TI - The validity of death certificates: routine validation of death certification and its effects on mortality statistics. AB - The 3478 death certificates (7.1% of all annual death certificates) of this study comprise those national death certificates in 1995 submitted for validation to the panel representing both medical and nosological expertise. As such, it is highly selected and represents, from the nosological point of view, the most inconsistently filled-in portion of Finnish death certificates. The routine validation procedure is essentially based on exploitation of the extra medical information, i.e. the case history, on the Finnish death certificate form. Altogether, 2813 (80.9%) out of 3478 certificates could be adjusted at the primary panel session; the rest required further clarification. The re-assignment of cause of death by the panel and the impact of panel adjustments on the national mortality statistics is assessed here by comparing the initial death certification and the finally registered underlying cause of death grouped into ICD-9 major categories with special reference to the subcategories of neoplasm, cardiovascular disease (HVD) and unnatural death. A statistically significant decline (p<0.0001) in deaths, both in the category of symptoms, signs and ill defined conditions and in the pulmonary circulation disease subcategory of HVD with 37.6 and 35.1%, respectively, was observed. The decrease of 11.1% in the benign or NUD neoplasm subcategory and the increase of 8.6 and 7.0% in the categories of endocrine disease, and musculo-skeletal and connective tissue disease, respectively, are essential observations as to the quality of the cause of death register. The effect on the HVD major category was practically nil. At the HVD-subcategorial level, a decrease of 14.0% for diseases of the veins and lymphatics and other circulatory diseases and an increase of 3.5% for hypertensive diseases (HYP) were the two next most obvious alterations to the diseases of the pulmonary circulation, but were without statistical significance. For ischaemic heart disease and other subcategories, the effects were minor. The unnatural deaths as a whole increased in the final statistics with only 0.9%. In the study data, categorial changes ranged from the decrease of 75.2% for symptoms, signs and ill-defined conditions to the increase of 77.3% for endocrine diseases. In conclusion, the Finnish death certificate form, death certification practices and cause of death validation procedure seem to serve the coding of causes of death for mortality statistics appropriately. The results of the study form a relevant reference background to evaluation of epidemiological studies on mortality. PMID- 11056268 TI - Death from thermal effects and burns. AB - One hundred and fifteen unselected autopsy cases of death from thermal effects and/or fire between 1990 and 1999 were analyzed with regard to time of death, signs of vitality at the scene of the fire, the manner and cause of death, and the extent of soft tissue loss. The cases represented approximately 6% of all autopsy cases at the Institute of Legal Medicine responsible for a catchment area with approximately 700,000 inhabitants. In 23 victims suffering burn injuries, death occurred during initial medical care or clinical treatment. The causes of death were primary respiratory arrest due to smoke poisoning or delayed shock caused by thermal injuries to the skin. Death occurred at the scene of the fatal event in 85 cases: eight cases exhibited no thermal effects; the cause of death in one of these cases was polytrauma incurred in a leap from a height; in seven cases there was poisoning due to smoke inhalation. The remaining 77 cases were characterized by signs of intensive thermal and/or fire effects. Clear signs of vitality (carboxyhemoglobin (COHb) in blood, inhalation and/or swallowing of soot) were found in 84.7% of the victims dying at the site of the fatal event. Of the 13 victims showing no signs of vitality at the scene, a cause of death could be determined in only seven cases; death in the other six cases remains unexplained. Quantification of the soft tissue loss revealed a possible correlation with the temperature and time course of heat exposure. PMID- 11056269 TI - In common batch searching of illicit heroin samples--evaluation of data by chemometrics methods. AB - In this study three illicit heroin samples which belonged to three different batches were subdivided into eight samples each. To simulate the dealers' chain, and to check the influence (if any) of diluents on the analytical results, some of the samples were cut with the most frequently used cutting substances, whereas the others were left unchanged. Samples were analysed (within a 10-week period of time) by gas chromatography-mass spectrometry (GC-MS) and characterised by seven variables each. To recover batch links among investigated heroin samples three multivariate methods, i.e. hierarchical clustering (HCA), principal component analysis (PCA) and k-nearest neighbours (k-NN), were applied on to the normalised and scaled analytical dataset. The classification abilities of the HCA, PCA and k NN were in the range from 95 to 100%. Disturbing effects due to the dilution of samples have not been observed. PMID- 11056270 TI - Synthesis of 4-methyl-5-arylpyrimidines and 4-arylpyrimidines: route specific markers for the Leuckardt preparation of amphetamine, 4-methoxyamphetamine, and 4 methylthioamphetamine. AB - General synthetic routes to 4-methyl-5-arylpyrimidines and 5-arylpyrimidines are described. 4-Benzylpyrimidine, 4-methyl-5-phenylpyrimidine, 4-(4 methoxybenzyl)pyrimidine, and 4-methyl-5-(4-methoxyphenyl)pyrimidine have been positively identified as route-specific by-products in the Leuckardt preparations of amphetamine and 4-methoxyamphetamine. Using headspace solid phase microextraction (SPME) 4-(4-methoxybenzyl)pyrimidine and 4-methyl-5-(4 methoxyphenyl)pyrimidine have been identified in illicit tablets containing 4 methoxyamphetamine. This is an indication that illicit laboratories use the Leuckardt method for the preparation of 4-methoxyamphetamine. Flatliner tablets containing 4-methylthioamphetamine have been screened for the presence of 4-(4 methylthiobenzyl)pyrimidine and 4-methyl-5-(4-methylthiophenyl)pyrimidine using both headspace and aqueous phase SPME. As these pyrimidines were not detected it would appear likely that illicit laboratories are not using the Leuckardt method for the preparation of 4-methylthioamphetamine. PMID- 11056271 TI - Fingermarks detection by eosin-blue dye. AB - Eosin-blue (I) dye, along with a phase transfer catalyst, has been used to detect latent fingerprints on a wide range of surfaces, including paper, glass, steel, lamination sheets, polythene, plastic and bakelite. PMID- 11056272 TI - Vacuum metal deposition: factors affecting normal and reverse development of latent fingerprints on polyethylene substrates. AB - Vacuum metal deposition (VMD) is an established technique for the development of latent fingerprints on non-porous surfaces. VMD has advantages over cyanoacrylate fuming, especially in circumstances where prints are old, have been exposed to adverse environmental conditions, or are present on semi-porous surfaces. Under normal circumstances, VMD produces 'negative' prints as zinc deposits onto the background substrate and not the print ridges themselves. A phenomenon of 'reverse' development, when zinc deposits onto the print ridges and not the background, has been reported by many authors but its causes have not been conclusively identified. Four plastic substrates were used in this study and these could be easily divided into two groups based on the types of development observed as the amount of deposited gold was increased. On group I plastics, identified as low-density polyethylene (LDPE), normal development then reverse development and finally no development resulted with increasing gold. On group II plastics, identified as high-density polyethylene (HDPE), normal development then over-development and finally poor-quality normal development resulted with increasing gold. Our results suggest that the difference between these plastic types causes variations in the gold film structure which in turn dictates the nature of the zinc deposition. On group I plastics, the structure and thickness of the gold film has been identified as the critical factor in the occurrence of normal or reverse development. Thin gold films on plastic substrates form small 'clusters' (or agglomerates) rather than the atoms being uniformly spread over the surface. The size and shape of these clusters is critical. Once the clusters reach a certain morphology, they no longer act as nucleation sites for zinc, and hence, zinc will not deposit onto the substrate. On group II plastics, results suggest that the gold clusters are smaller and more densely packed. Hence, even though the same amount of gold has been deposited, the gold clusters in this case do not reach the critical morphology and so continue to act as nucleation sites for zinc.Typically, zinc will not deposit onto the fingerprint ridges as the gold nucleation sites are buried within the print residue. However, when more gold is deposited, gold emerges at the surface of the latent print allowing zinc deposition onto the ridges. The rate of gold evaporation was found not to affect the structure of the gold film, although a slower rate of evaporation resulted in more effective deposition. PMID- 11056273 TI - On the diagnosis of hypoglycemia in car drivers--including a review of the literature. AB - Hypoglycemia may reach forensic relevance concerning the psychophysical ability of running a car and with respect to a possible imputability. Our retrospective study included anamnesis and clinical symptoms observed in drivers with impaired performance to point out correlations between biochemical parameters and the actual course of the disease and its sequelae on roadworthiness. Clinical biochemical estimations were performed on glucose, lactic acid and ethanol including a toxicological screening. It is proposed to utilize the estimation of glucose and lactic acid (the so-called combined value) in blood samples taken on behalf of the police as an important hint to the actual state of glucose metabolism. The anamnesis and the symptomatology may complete the biochemical analyses. PMID- 11056274 TI - Regional Italian allele frequencies at nine short tandem repeat loci. AB - An Italian population study was performed on the loci D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317, D7S820 and a portion of the X-Y homologous gene Amelogenin for gender determination using the AmpFlSTR Profiler kit (PE Biosystems, Foster city, CA). This study was done on a population of 618 unrelated Italian individuals from 18 regions in Italy (except for Valle d'Aosta and Sardinia) to determine allele frequencies for each STR locus, and to evaluate STR technology for developing an Italian Offender DNA database. PMID- 11056275 TI - A repository of 14 PCR-loci Italian gene frequencies in the World Wide Web. AB - A collection of 6830 typing results produced by the Immunohematology Laboratory at the UCSC, pertaining to 11 STRs (FES/FPS, vWA31, HUMTH01, F13A1, MBP, D21S11, D7S460, D18S51, CD4, TPOX, CSF1PO) and 3 AmpFLPs (D1S80, APO-B, COL2A1), is publicly available as an electronic archive at a website. PMID- 11056276 TI - Genetic polymorphism at three STR loci--CSF1PO, HUMTHO1 and TPOX, and the AMP-FLP locus D1S80 for the Chinese population in Hong Kong. AB - Allele frequencies for three STR loci, namely, CSF1PO, HUMTHO1 and TPOX, and the AMP-FLP locus D1S80 were obtained from a sample of 351 unrelated Chinese in Hong Kong. PMID- 11056277 TI - STR data for the AmpFlSTR SGM plus from Slovenia. AB - Allele frequencies for the 10 STRs loci included in the AmpFISTR SGM Plus kit (PE Applied Biosystem) were obtained from a sample of 321 unrelated individuals born in Slovenia. PMID- 11056278 TI - STR data from Basque country autochthonous population. PMID- 11056279 TI - Thai population data on nine tetrameric STR loci - D3S1358, vWA, FGA, TH01, TPOX, CSF1PO, D5S818, D13S317 and D7S820. AB - Allele frequencies for the nine STR loci included in the AmpFlSTR Profiler kit were determined in a Thai population of 100 unrelated individuals. PMID- 11056280 TI - Population data on D16S539, D7S820, D13S317, LPL, F13B and D1S80 loci in a sample of Caucasian-Mestizos from Colombia. PMID- 11056281 TI - STR data (AmpFlSTR profiler plus) from north Portugal. AB - Allele frequencies for the nine STRs included in the AmpFlSTR Profiler Plus kit (D3S1358, VWA, FGA, D8S1179, D21S11, D18S51, D5S818, D13S317 and D7S820) were estimated from a sample of 365-427 unrelated individuals born in north Portugal. PMID- 11056282 TI - The ratio of insulin to C-peptide can be used to make a forensic diagnosis of exogenous insulin overdosage. AB - A 25-year-old man was killed by his lover by an intravenous injection of insulin and then air. We had difficulty in determining whether insulin had really been injected, so we have 18 control cases and proved that the ratio of insulin to C peptide in a corpse can be used as positive evidence for the insulin overdosage. PMID- 11056284 TI - Argemone mexicana poisoning: autopsy findings of two cases. AB - Epidemic dropsy, a disease due to Argemone mexicana poisoning, is characterized by pathological accumulation of diluted lymph in body tissues and cavities. Recently, the largest epidemic of the disease in India affected Delhi and its neighboring states during the months of August-September 1998. Over 3000 persons fell ill, and more than 65 died in the state of Delhi alone. Two cases belonging to the same family died, out of the large number of cases admitted in this tertiary care teaching hospital situated in eastern part of Delhi. Autopsy findings of these two cases are presented and discussed here along with the review of toxicity due to this poisoning. PMID- 11056285 TI - Burn injuries caused by a hair-dryer--an unusual case of child abuse. AB - About 1.4-26% burn injuries in children appear to be abusive in origin. A 2.5 year-old girl was referred to our institute because of suspected child abuse. Clinical examination and later interrogation of the mother revealed non-recent deep second degree burn injuries on both gluteal regions, caused by the partner of the mother by pressing a hand-held hair-dryer against the skin. The authors present the findings of this unusual method of child abuse. PMID- 11056286 TI - Prolonged QT interval and sudden infant death--report of two cases. AB - In the two cases where infants died suddenly and unexpectedly the electrocardiogram (ECG) of a younger sibling (case 1) and of a living twin (case 2) led to the suspicion that the two infants could have died from long QT syndrome (LQTS). In case 1, a His bundle (HB) dispersion and a pronounced hypoplasia of the right external nucleus arcuatus were detected. In case 2, a severe interstitial pneumonia and an accompanying mild myocarditis were found by histology. Molecular genetic investigations of the coding regions of the genes, HERG, KVLQT1 and SCN5A gave no indication for the mutations, thus, affecting related myocardial ion channels as possible sources of inhomogeneity of repolarisation. Since a molecular genetic deviation could not yet be elaborated the possible role of related disturbance remains unknown. PMID- 11056287 TI - An autopsy case of neuroleptic malignant syndrome (NMS) and its immunohistochemical findings of muscle-associated proteins and mitochondria. AB - Neuroleptic malignant syndrome (NMS) is a rare but potentially fatal disorder. In forensic cases, post-mortem diagnosis of NMS is sometimes difficult if ante mortem information, such as neuroleptic ingestion or signs and symptoms, cannot be obtained. A 39-year-old Japanese male on a neuroleptic treatment regimen suddenly became agitated and died. Autopsy revealed muscle rigidity and hyperthermia. Post-mortem examination of blood revealed elevation of creatine phosphokinase-MM (CK-MM) and lactate dehydrogenase-4 and dehydrogenase-5 (LDH-4 and LDH-5). In renal glomeruli and tubules, myoglobin was stained immunohistochemically. From these findings, the cause of death was considered to be NMS. To support the diagnosis of NMS, both skeletal and cardiac muscles were stained with actin, myoglobin, desmin and mitochondria antibodies immunohistochemically. Actin, myoglobin, desmin, and mitochondria had been lost from skeletal, but not from the cardiac muscle, which suggested that only the skeletal muscle was damaged. Moreover, because mitochondria had disappeared only from the skeletal muscle, it was considered that skeletal muscle degeneration was caused by mitochondrial damage. Therefore, it is suggested that immunostaining of skeletal muscle by antibodies for muscle-associated proteins and mitochondria is useful to corroborate a diagnosis of NMS. PMID- 11056288 TI - A deoxyinosine specific endonuclease from hyperthermophile, Archaeoglobus fulgidus: a homolog of Escherichia coli endonuclease V. AB - Deoxyadenosine undergoes spontaneous deamination to deoxyinosine in DNA. Based on amino acids sequence homology, putative homologs of endonuclease V were identified in several organisms including archaebacteria, eubacteria as well as eukaryotes. The translated amino acid sequence of the Archaeoglobus fulgidus nfi gene shows 39% identity and 55% similarity to the E. coli nfi gene. A. fulgidus endonuclease V was cloned and expressed in E. coli as a C-terminal hexa-histidine fusion protein. The C-terminal fusion protein was purified to apparent homogeneity by a combination of Ni(++) affinity and MonoS cation exchange liquid chromatography. The purified C-terminal fusion protein has a molecular weight of about 25kDa and showed endonuclease activity towards DNA containing deoxyinosine. A. fulgidus endonuclease V has an absolute requirement for Mg(2+) and an optimum reaction temperature at 85 degrees C. However, in contrast to E. coli endonuclease V, which has a wide substrate spectrum, endonuclease V from A. fulgidus recognized only deoxyinosine. These data suggest that the deoxyinosine cleavage activity is a primordial activity of endonuclease V and that multiple enzymatic activities of E. coli endonuclease V were acquired later during evolution. PMID- 11056289 TI - Mutation in recR gene of Deinococcus radiodurans and possible involvement of its product in the repair of DNA interstrand cross-links. AB - We previously reported that some Deinococcus radiodurans mutants are sensitive to DNA interstrand cross-linking agents but resistant to UV and gamma-rays. We isolated DNA fragments from a D. radiodurans genomic library which complemented the mitomycin C sensitivity of one of these mutants. One 3.2kb-long fragment contains an open reading frame of approximately 700bp and the deduced amino acid sequence is very homologous to other prokaryotic RecR proteins. This open reading frame in the mitomycin C-sensitive mutant strain contains a frame shift mutation at its carboxyl terminal region. These data suggest that RecR protein plays an important role in the resistance to interstrand cross-links in this bacterium. PMID- 11056290 TI - The influence of combined Fpg- and MutY-deficiency on the spontaneous and gamma radiation-induced mutation spectrum in the lacZalpha gene of m13mp10. AB - One of the most predominating oxidative DNA damages, both spontaneously formed and after gamma-radiation is 7, 8-dihydro-8-oxoguanine (8oxoG). This 8oxoG is a mutagenic lesion because it can mispair with adenine instead of the correct cytosine leading to G:C to T:A transversions. In Escherichia coli (E. Coli) base excision repair (BER) is one of the most important repair systems for the repair of 8oxoG and other oxidative DNA damage. An important part of BER in E. coli is the so-called GO system which consists of three repair enzymes, MutM (Fpg), MutY and MutT which are all involved in repair of 8oxoG or 8oxoG mispairs. The aim of this study is to determine the effect of combined Fpg- and MutY-deficiency on the spontaneous and gamma-radiation-induced mutation spectrum of the lacZalpha gene. For that purpose, non-irradiated or gamma-irradiated double-stranded (ds) M13mp10 DNA, with the lacZalpha gene inserted as mutational target sequence was transfected into an E. coli strain which is deficient in both Fpg and MutY (BH1040). The resulting mutation spectra were compared with the mutation spectra of a fpg(-) E. coli strain (BH410) and a wild type E. coli strain (JM105) which were determined in an earlier study. The results of the present study indicate that combined Fpg- and MutY-deficiency induces a large increase in G:C to T:A transversions in both the spontaneous and gamma-radiation-induced mutation spectra of BH1040 (fpg(-)mutY(-)) as compared to the fpg(-) and the wild type strain. Besides the increased levels of G:C to T:A transversions, there is also an increase in G:C to C:G transversions and frameshift mutations in both the spontaneous and gamma-radiation-induced mutation spectra of BH1040 (fpg(-)mutY( )). PMID- 11056291 TI - DNA repair capacity: inconsistency between effect of over-expression of five NER genes and the correlation to mRNA levels in primary lymphocytes. AB - We have previously shown that high DNA repair capacity protects psoriasis patients against chemically induced basal cell carcinoma [Dybdahl et al. Mutat. Res. 433 (1999) 15-22]. We have used the same study persons to investigate the correlation between expression of eight genes involved in nucleotide excision repair and DNA repair capacity. mRNA levels of XPA, XPB, XPC, XPD, XPF, XPG, CSB and ERCC1 in primary lymphocytes from 33 individuals were quantified by dot-blots and normalized to beta-actin. ERCC1 and XPD mRNA quantities were highly correlated (r=0.89; P<10(-11)) while XPA, XPB, XPC, XPG, XPFand CSB mRNAs were moderately correlated (r=0.2-0.7). Thus, the mRNA expressions seem to fall in at least two groups. There was a three to sevenfold variation in the expression levels of the mRNAs. This is in contrast to the more than a hundredfold variation in mRNA levels reported in cancer patients.DNA repair capacity was measured in a host cell reactivation assay, where primary lymphocytes were transfected with an UV-irradiated plasmid encoding firefly-luciferase. Only ERCC1 and XPD mRNA levels correlated with the DNA repair capacity (P<0.03). In order to see if ERCC1 or XPD activity was limiting for DNA repair, we cotransfected with plasmids encoding NER genes, thus over-expressing either XPB, XPC, XPD, CSB or ERCC1 in the host cell reactivation assay. Only XPB over-expression increased DNA repair capacity. Thus, there is no indication that neither XPD nor ERCC1 limits the DNA repair capacity. However, our results indicate that ERCC1 and XPD mRNA levels may be used as a proxy for DNA repair capacity in lymphocytes. PMID- 11056292 TI - Synergistic DNA damaging effects of 4-nitroquinoline-1-oxide and non-effective concentrations of methyl methanesulfonate in human fibroblasts. AB - DNA damage and DNA repair in human fibroblasts induced by the combination mixture of the genotoxic agents methyl methanesulfonate (MMS) and 4-nitroquinoline-1 oxide (4-NQO) were studied using the comet assay and the unscheduled DNA synthesis (UDS), respectively. Cells were simultaneously treated for 1h with the no observed effect concentration (noec) of MMS and increasing concentrations of 4 NQO or vice versa. Different results were obtained with the two types of mixtures. When the noec of 4-NQO was combined with increasing concentrations of MMS, no combination effects were observed. However, in experiments with increasing concentrations of 4-NQO and the noec of MMS, an increase in DNA damage and repair (and an enhancement of cytotoxicity) was demonstrated. Quantitative analysis of the effects by the isobologram method confirmed synergistic responses in both tests. We are proposing interactive actions between 4-NQO and MMS, whereby 4-NQO facilitates the attack of MMS on the DNA bases. PMID- 11056293 TI - Inhibition of DNA synthesis is a potent mechanism by which cytostatic drugs induce homologous recombination in mammalian cells. AB - Recombination is a process thought to be underlying genomic instability involved in carcinogenesis. This report examines the potential of cytostatic drugs to induce intrachromosomal homologous recombination. In order to address this question, the hprt gene of a well-characterized mammalian cell line was employed as a unique endogenous marker for homologous recombination. Commonly used cytostatic drugs with different mode of action were investigated in this context, i.e. bifunctional alkylating agents, inhibitors of DNA synthesis, inhibitors of topoisomerases and a spindle poison. With the exception of the spindle poison, all these drugs were found to induce homologous recombination, with clear differences in their recombination potency, which could be related to their mechanism of action. Bifunctional alkylating agents were the least efficient, whereas inhibitors of DNA synthesis were found to be the most potent inducers of homologous recombination. This raises the question whether these later drugs should be considered for adverse effects in cancer chemotheraphy. PMID- 11056294 TI - Paternal exposure to cyclophosphamide induces DNA damage and alters the expression of DNA repair genes in the rat preimplantation embryo. AB - Chronic low dose treatment of male rats with cyclophosphamide, an anticancer alkylating agent, damages male germ cells, resulting in greater than 80% peri implantation progeny loss. Little transcription or repair takes place in the DNA of post-meiotic male germ cells. The spermatozoal genome regains its transcriptional capacity in the fertilized oocyte. We hypothesized that as a consequence of exposure of male rats to cyclophosphamide DNA damage to the male genome is transmitted to the conceptus; furthermore, this damage leads to alterations in the expression profiles of DNA repair genes during preimplantation development. Male rats were treated with either saline or cyclophosphamide (6mg/kg/day, 4-6 weeks) and mated to control females; 1-8 cell stage embryos were collected. The alkaline comet assay was used to assess DNA damage in 1-cell embryos. A significantly higher percentage (68%) of the embryos fertilized by cyclophosphamide-exposed spermatozoa displayed a comet indicative of DNA damage, compared to those sired by control males (18%). The in situ transcription/antisense RNA approach was used to determine if DNA damage alters the expression of DNA repair genes in early embryos. Dramatic increases in the transcripts for selected members of the nucleotide excision repair family (XPC, XPE and PCNA), mismatch repair family (PMS1), and recombination repair family (RAD50) were found in 1-cell stage embryos sired by cyclophosphamide-treated males compared to controls, while decreases in the expression of base excision repair family members (UNG1, UNG2, and XRCC1) and in recombination repair transcripts (RAD54) were observed. By the 8-cell stage, transcripts for specific members of the nucleotide excision repair family (XPC) and mismatch repair family (MSH2, PMS2) were elevated greatly in control embryos compared to embryos sired by drug-treated males; in contrast, transcripts for other members of the nucleotide excision repair family (XPE, PCNA), as well as some of the base excision repair family (UNG1), were elevated in embryos sired by drug-treated males. Therefore, DNA damage incurred in spermatozoa, following cyclophosphamide exposure is associated with alterations in the expression profiles of DNA repair genes in preimplantation embryos as early as the 1-cell stage. Genotoxic stress may disturb the nuclear remodeling and reprogramming events that follow fertilization and precede zygotic genome activation. PMID- 11056295 TI - Sensitivity of a S. cerevisiae RAD27 deletion mutant to DNA-damaging agents and in vivo complementation by the human FEN-1 gene. AB - We have investigated the sensitivity to DNA-damaging agents of a strain of Saccharomyces cerevisiae containing a deletion of the RAD27 gene. The mutant strain is sensitive to a number of alkylating agents that modify DNA at a variety of positions, including one that produces primarily phosphotriesters. In contrast, the mutant strain is not sensitive to the oxidizing agent hydrogen peroxide. The introduction of a plasmid containing the FEN-1 gene (the human ortholog of the RAD27 gene) can substantially complement the sensitivity to alkylating agents observed in the mutant strain. PMID- 11056296 TI - The oestrogen receptor and its selective modulators in gynaecological and breast cancer. PMID- 11056297 TI - Oestrogen and female cancers. the past 100 years. PMID- 11056298 TI - Molecular mechanisms of oestrogen - the gynaecologists' viewpoint. PMID- 11056299 TI - New insights in oestrogen receptor (ER) research - the ERbeta. PMID- 11056300 TI - Structural insights into the mechanisms of agonism and antagonism in oestrogen receptor isoforms. AB - Here we summarise the results that have emerged from our structural studies on the oestrogen receptor (ER) ligand-binding domain. We have investigated the conformational effects of a variety of ligands on the structures of both ER isoforms. Each class of ligand (agonists, partial agonists and selective oestrogen receptor modulators) induces a unique conformation in the receptor's ligand-dependent transcriptional activation function. Together these studies have broadened our understanding of ER function by providing a unique insight into ER's ligand specificity and the structural changes that underlie receptor agonism and antagonism. PMID- 11056301 TI - Detection of the oestrogen receptor (ER). immunohistochemical versus cytosol measurements. AB - Oestrogen receptor (ER) content gives a direct indication of the chances that a breast cancer patient will show a sustained response to endocrine therapy. Thus, an ER value should be recorded for every breast cancer patient. ER was traditionally measured by a ligand binding assay (LBA). LBA is not suitable for all routine hospitals in which breast cancer is treated. More appropriate is immunohistochemistry (IHC). This paper identifies advantages and disadvantages of both assays, suggests that both methods predict equally response to endocrine therapies and describes a simple, semi-quantitative IHC for which external quality assurance works successfully. PMID- 11056302 TI - The immunocytochemical versus cytosol measurement of the oestrogen receptor in invasive breast cancer tissue. AB - To compare two methods of measurement of oestrogen receptor (ER)-expression in invasive breast cancer tissue. Sections from 299 breast cancer cases were stained for the ER by immunocytochemical assay (ICA), using mouse monoclonal antibody (MAb) NCL-ER-6SF11, and by the dextran-coated charcoal assay (DCC). Concordant results were observed in 230 of the 299 cases (77%), 69 patients had discordant results (kappa=0.537). We found a moderate concordance between ICA and DCC for ER measurement in breast cancer tissue. If we change the golden standard from DCC to ICA, 23% of patients would receive a different therapeutic approach. PMID- 11056303 TI - How best to express oestrogen receptor activity. AB - Oestrogen receptor (ER) activity, detected and expressed in a variety of ways, is important in breast cancer. Experience in Edinburgh (1973-1996) showed that [1] no single mode of expression was entirely satisfactory, [2] the probability of a good 'outcome' (prognosis or response to endocrine therapy) increased with increasing activity (either fmol ER sites/mg protein or per cent cells staining for ER). Thus the use of a single 'cut-off' should be avoided and activity quantified, or stratified into categories. PMID- 11056304 TI - Oestrogen receptor expression in normal breast epithelium. AB - Higher levels of oestrogen receptor (ER) expression in normal breast epithelium may compound the increase in breast cancer risk seen with prolonged estrogen exposure. In prior studies, we have used immunohistochemical ER assays on fresh frozen samples of benign breast tissue. Future studies will be more feasible on paraffin-embedded samples, and newer, more sensitive antibodies are now available. We examined 30 samples of paraffin-embedded breast epithelium from postmenopausal women with two antibodies, 6F11 and TE111. We find that the median labelling indices for ER are significantly higher using these antibodies, compared with previous results. The threshold for ER positivity will, therefore, have to be reset in future studies, since there are still many issues that remain to be resolved in this area. PMID- 11056305 TI - P27(KIP1) expression indicates that steroid receptor-positive cells are a non proliferating, differentiated subpopulation of the normal human breast epithelium. AB - To test the hypothesis that steroid receptor-expressing cells are derived from the proliferative population, we examined expression of the p27(KIP1) inhibitor of cyclin-dependent kinase activity (a differentiation marker) while tracking the fate of proliferating cells in normal human breast tissue implanted into athymic nude mice using tritiated thymidine [3H]-dT. We identified a small number of cells that appeared to have divided just once before switching on p27(KIP1) expression. p27(KIP1)+ve cells also expressed steroid receptors, but not the Ki67 proliferation-associated antigen. These data support the hypothesis that steroid receptor-expressing cells are a differentiated population within the normal human breast epithelium. PMID- 11056306 TI - The oestrogen receptor (ER) in normal and abnormal uterine tissue. AB - Glandular epithelium and stroma of the endometrium show typical behavioural patterns in the expression of oestrogen receptors (ERs) due to both endogenous and exogenous hormonal influence. Thus, the ER is increasingly expelled under the influence of oestrogen during the first half of the cycle. Under the influence of progesterone, the ER disappears during the luteal phase and is not even detectable after day 21. During menopause, the atrophic endometrium typically shows very little, if any, ER expression. In cases of oestrogen-induced hyperplasias, the receptor can again be demonstrated. The ER only disappears when nuclear irregularities occur in cases of adenomatous hyperplasia. In cases of invasive carcinoma, a heterogenous picture is seen which closely correlates with the degree of differentiation. PMID- 11056307 TI - The oestrogen receptor (ER) in vulva, vagina and ovary. AB - The oestrogen receptor (ER) has been identified in normal and neoplastic epithelia of the vulva, vagina and ovary using biochemical, immunohistochemical (IHC) and molecular techniques. Its presence has not translated into effective antineoplastic therapy for malignancies arising from these sites. PMID- 11056308 TI - Steroid receptors and metastatic potential in endometrial cancers. AB - The relative overexpression of oestrogen receptor (ER)-alpha exon 5 splicing variant (ER-alpha E5SV), ER-beta and progesterone receptor (PR) from B (PR-B) without transcriptional repression by PR from A (PR-A) might be related to the metastatic potential and partially cause deviation from sex steroidal dependency in endometrial cancers. PMID- 11056309 TI - Modulation of oestrogen action by receptor gene inhibition. AB - Selective oestrogen receptor downregulators (SERDs) are a class of highly effective steroidal antitumour agents that reduce cellular levels of the oestrogen receptor (ER). In this study, we compared the efficacy by which three novel molecular approaches: (1) antisense oligonucleotides; (2) antisense RNA; and (3) dominant negative mutants are able to act as SERDs. Using transient and, where appropriate, stable gene transfection experiments we found that constitutive overexpression of ER antisense RNA and a hormone-binding domain compromised dominant-negative ER mutant (DNER-1), were most effective at downregulating ER expression and/or activity in vitro. PMID- 11056310 TI - Identification of women at high risk of developing endometrial cancer on tamoxifen. PMID- 11056311 TI - Monitoring for endometrial disorders in 406 breast cancer women treated by tamoxifen. a low aggressive strategy. PMID- 11056312 TI - Absence of correlation between risk factors for endometrial cancer and the presence of tamoxifen-associated endometrial polyps in postmenopausal patients with breast cancer. AB - In order to investigate the presence of established risk factors for endometrial carcinoma in postmenopausal patients with breast cancer and with tamoxifen associated endometrial polyps we compared a group of 25 patients with tamoxifen associated endometrial polyps with 25 tamoxifen-treated patients without endometrial polyps. No significant differences were found between both groups of patients in age, parity, time after breast cancer and after menopause, duration and daily and total cumulative dose of tamoxifen intake, body mass index and serum levels of luteinising hormone (LH), follicle-stimulating hormone (FSH), oestradiol (E2), progesterone, sex hormone-binding globulin (SHBG), tamoxifen and CA125. So far there is no evidence that these polyps are premalignant lesions. PMID- 11056313 TI - Modulation of endometrial transforming growth factor beta (TGFbeta) by tamoxifen. AB - Transforming growth factor beta (TGFbeta) immunoreactivity was determined in endometria from non-drug-therapy and tamoxifen-treated patients. Sections were scored for pathology and quantity image analysis performed to determine levels of glandular- or fibrosis-associated TGFbeta1. Tamoxifen-treated patients displayed greater levels of endometrial dysplasia and glandular hyperplasia, in addition to a statistically significant (P<0.0001) elevation in gland-associated TGFbeta1 protein. PMID- 11056314 TI - Inhibition of oestrogen receptor activity by the co-repressor HET/SAF-B is relieved by blockade of histone deacetylase activity. PMID- 11056315 TI - Does tamoxifen change oestrogen and progesterone receptor expression in the endometrium and breast? AB - We studied the expression of oestrogen and progesterone receptors (ER, PR) in postmenopausal women receiving tamoxifen for breast cancer. In addition the literature addressing the question of ER and PR expression in breast tissue during treatment with tamoxifen was reviewed. We demonstrated consistent expression of ER and PR in endometria from patients receiving tamoxifen, with a trend towards a higher proportion of receptor positive specimens during tamoxifen. In breast cancer tissue, the ER content seemed to be reduced following tamoxifen treatment. After short time exposure to tamoxifen, the PR appeared to be increased, longer treatment caused the PR to go down to pretreatment levels or below. PMID- 11056316 TI - Breast cancer prevention: results of the National Surgical Adjuvant Breast and Bowel Project (NSABP) breast cancer prevention trial (NSABP P-1: BCPT). PMID- 11056317 TI - Update on tamoxifen to prevent breast cancer. The Italian Tamoxifen Prevention Study. PMID- 11056318 TI - A brief review of the breast cancer prevention trials. PMID- 11056319 TI - Breast cancer prevention: the next steps. PMID- 11056320 TI - Update on raloxifene to prevent endometrial-breast cancer. AB - In the mid 1980s when tamoxifen was shown to be associated with endometrial neoplasia there was a renewed interest in another SERM compound, raloxifene. Experimental animal data suggested that raloxifene did not stimulate the endometrium as tamoxifen does while having similar anti-oestrogenic effects in breast tissue as tamoxifen. Clinical data has now shown that raloxifene does not stimulate the endometrium in postmenopausal women. It results in no hyperplasia, no increase in endometrium thickness or polyp formation and virtually no proliferation. Further studies are necessary to see if long-term raloxifene use will reduce the risk of endometrial cancer. In studies of raloxifene as treatment for osteoporosis, when viewed as a secondary endpoint there was a significant reduction in risk of new onset breast cancer. Further studies with breast cancer as a primary endpoint are ongoing (the STAR Trial). PMID- 11056321 TI - Tamoxifen in the treatment of recurrent ovarian carcinoma. AB - This review examines the evidence for useful clinical activity of tamoxifen in women with ovarian carcinoma who have failed conventional cytotoxic chemotherapy. The optimised search strategy of the Cochrane Gynaecological Cancer Collaborative Review Group by Williams was used. Tamoxifen demonstrates a modest degree of effectiveness in ovarian cancer refractory to cytotoxic chemotherapy. The overall objective response rate in all trials (647 patients) was approximately 11%. There is, however, a wide variation in the objective response rates in the different trials (0-56%). Tamoxifen therapy has limited efficacy in refractory ovarian carcinoma. However, considering the mild toxicity of tamoxifen, occasional long term palliation and lack of alternatives, the drug has a useful role in heavily pretreated patients with ovarian cancer. PMID- 11056322 TI - Toremifene. where do we stand? AB - Toremifene is a chlorinated triphenylethylene that is indicated for postmenopausal breast cancer. For advanced disease, toremifene has been found to be as effective and at least as well tolerated as tamoxifen. The same appears to apply for adjuvant setting. After a total cumulative clinical exposure to toremifene of approximately 140000 patient-years, only 9 cases of endometrial carcinoma have been reported. The annual hazard rate (per 1000 patient-years) of developing endometrial carcinoma in breast cancer patients on adjuvant toremifene is 1.14 (versus tamoxifen 2.0 and placebo 0.4). Although toremifene (being a partial agonist) may unmask pre-existing endometrial tumours, there is no clinical data implying that it would per se cause endometrial carcinoma. PMID- 11056323 TI - The selective oestrogen receptor modulators idoxifene and raloxifene have fundamentally different cell-specific oestrogen-response element (ERE) dependent/independent mechanisms in vitro. AB - Idoxifene and raloxifene are selective oestrogen receptor modulators (SERMs) that by definition exhibit tissue-specific agonist or antagonist properties via interactions with the oestrogen receptor (ER). Idoxifene acts as an oestrogen agonist in osteoblastic cells via an ER/ERE-mediated mechanism. In contrast, raloxifene is an antagonist via the ERE in osteoblastic cells. Like the pure antagonist ICI 182,780, raloxifene inhibited the potent agonist activity of both 17beta-oestradiol and idoxifene through the ERE whereas idoxifene had no effect on the agonist activity of 17beta-oestradiol via the ERE. In breast cancer cells, both raloxifene and idoxifene were potent antagonists of ERE-mediated 17beta oestradiol action suggesting an ERE-dependent mechanism of action for both ligands in these cells. Therefore, these SERMs exhibit cell-specific ERE dependent and -independent mechanisms of action. PMID- 11056324 TI - Other selective oestrogen receptor modulators (SERMs) in development. PMID- 11056325 TI - The oestrogen receptor and its selective modulators in gynaecological oncology. PMID- 11056326 TI - Soyfoods and soybean phyto-oestrogens (isoflavones) as possible alternatives to hormone replacement therapy (HRT). PMID- 11056327 TI - Inhibition of tamoxifen's therapeutic benefit by tangeretin in mammary cancer. AB - Tangeretin, a molecule present in citrus fruits and in certain 'natural' menopausal medications, is an effective tumour growth and invasion inhibitor in vitro of human MCF 7/6 breast cancer cells. However, when added to the drinking water of MCF 7/6 tumour-bearing mice it neutralises the beneficial tumour suppressing effect of tamoxifen. Tangeretin reduces the number of natural killer cells. This may explain why the beneficial suppressive effect of tangeretin on MCF 7/6 cell proliferation in vitro is completely counteracted in vivo. PMID- 11056328 TI - How to manage the menopause following therapy for breast cancer. is raloxifene a safe alternative? AB - Raloxifene is a selective oestrogen receptor modulator (SERM) that has anti oestrogenic effects on breast and endometrial tissue and oestrogenic actions on bone, lipid metabolism and blood clotting. In postmenopausal women raloxifene decreases bone turnover and increases bone mineral density, reducing the incidence of vertebral fractures. Unlike tamoxifen, raloxifene does not cause endometrial hyperplasia or cancer, as demonstrated by endometrial monitoring with ultrasonography and biopsy during treatment. Evidence suggests that raloxifene lowers total low-density lipoprotein cholesterol levels behaving like oestrogens, but does not increase high-density lipoprotein cholesterol levels. In randomised clinical trials on postmenopausal women with osteoporosis, raloxifene reduced the risk of newly diagnosed ER-positive invasive breast cancer by 76% during a median of 40 months of treatment. However, raloxifene does not alleviate early menopausal symptoms, such as hot flushes and urogenital atrophy, and may even exacerbate some of them. In conclusion, raloxifene may be an alternative for the prevention of long-term effects of oestrogen deficiency (osteoporosis and heart diseases) in women with previous breast cancer not having hot flushes. For symptomatic patients, the association of raloxifene with different drugs which have demonstrated efficacy in the control of vasomotor symptoms is now under evaluation. PMID- 11056329 TI - Tibolone actions on normal and breast cancer cells. AB - Tibolone and its main derivatives were studied in an original model of cultures of normal human epithelial breast (HBE) cells on proliferation, differentiation and apoptosis, the three mechanisms responsible for breast homeostasis. Tibolone and its Delta4 isomer were antiproliferative, both in the absence and presence of oestradiol (E2). The oestrogenic 3alpha and 3beta hydroxy derivatives did not display any mitogenic activities in HBE cells. Moreover, at 1 microM, they were antiproliferative. Tibolone and its Delta isomer increased the 17beta hydroxysteroid dehydrogenase activity similarly to that observed with progestins [1]. Apoptosis was increased in HBE cells to a similar range as with the pure pregnane progestin, Org2058. We also studied the extent of apoptosis in hormone dependent breast cancer cell lines. Tibolone and its Delta4 isomer also increased apoptosis, especially in ZR75-1 cells containing progesterone and androgen receptors [2]. We could demonstrate that these pro-apoptotic actions of tibolone and its Delta4 isomer were mediated at least partially through the bcl-2-family of proteins. Moreover, the antiproliferative and pro-apoptotic activities of tibolone, as well as Org2058, were mediated by increasing catalase activities in breast cancer cells. Thus, in breast cells, tibolone slows down the proliferation rate, increases differentiation and apoptosis. These actions seem to be optimal on breast tissue. PMID- 11056330 TI - Clinico-pharmacological aspects of different hormone treatments. AB - During the last decade, several new drugs and classes of drugs have become available for breast cancer treatment. Thus, in addition to tamoxifen we have got several new selective oestrogen receptor modulators (SERMs) with a partially different pharmacological profile. The first generation aromatase inhibitor, aminoglutethimide, has been replaced by more potent and less toxic inhibitors belonging to the triazole class (anastrozole and letrozole) and, more recently, the steroidal aromatase inactivator exemestane [1-3]. These drugs have all revealed a better toxicity profile and, in general, an improved antitumour activity, compared with conventional therapy. Faslodex, the first representative of the so-called 'pure' oestrogen antagonists, has shown beneficial effects in patients resistant to tamoxifen [4]. PMID- 11056331 TI - An overview of the use of non-steroidal aromatase inhibitors in the treatment of breast cancer. AB - A number of potent and selective non-steroidal aromatase inhibitors are now available for the treatment of advanced breast cancer in postmenopausal women. In particular, anastrozole represents a significant advantage over earlier agents, such as aminoglutethimide and formestane, in terms of both efficacy and tolerability. These agents are now established as the second-line therapy of choice in postmenopausal women with advanced disease progressing on tamoxifen and, furthermore, data are now available on the efficacy and tolerability of anastrozole as first-line treatment of advanced breast cancer compared with tamoxifen. The full potential of the new-generation aromatase inhibitors in the treatment of breast cancer is currently being investigated in a large programme of clinical trials, including evaluation as neoadjuvant treatment in postmenopausal women with newly-diagnosed locally-advanced or large operable breast cancers, as first-line treatment of advanced breast cancer in postmenopausal women. Aromatase inhibitors have been available for over 20 years; the ability of these compounds to reduce circulating oestradiol levels has been shown to produce clinical benefit in postmenopausal women with advanced breast cancer. Early aromatase inhibitors, however, such as aminoglutethimide and formestane, were not specific for the aromatase enzyme and resulted in significant side-effects. PMID- 11056332 TI - Randomised study of anastrozole versus tamoxifen as first-line therapy for advanced breast cancer in postmenopausal women. AB - A total of 668 patients (340 anastrozole and 328 tamoxifen) were randomised in a double-blind, double-dummy multicentre study. Anastrozole was given in a dose of 1 mg once daily and compared with tamoxifen 20 mg daily in postmenopausal patients with tumours that were hormone-receptor positive or of unknown receptor status. The efficacy and tolerability of anastrozole was compared with that of tamoxifen as first-line therapy for advanced breast cancer. The median time to progression was similar for both treatments (8.2 months in anastrozole patients and 8.3 months in tamoxifen patients). Anastrozole was also as effective as tamoxifen in terms of objective response-rate with 33% in the anastrozole group and 32.6% in the tamoxifen group achieving a complete or partial response. Both treatments were well tolerated. However, incidences of thromboembolic events and vaginal bleeding were reported in fewer patients treated with anastrozole than with tamoxifen. In conclusion, these findings indicate that anastrozole can be considered as first-line therapy for postmenopausal women with advanced breast cancer. PMID- 11056333 TI - Exemestane improves survival compared with megoestrol acetate in postmenopausal patients with advanced breast cancer who have failed on tamoxifen. results Of a double-blind randomised phase III trial. AB - Exemestane is an aromatase inactivator. 769 Postmenopausal women with advanced breast cancer who had failed on tamoxifen were randomised to exemestane or megoestrol acetate in this double-blind trial. Objective response rate was similar between treatments. Median time to progression, time to treatment failure and overall survival was significantly longer with exemestane. Drug-related withdrawals and drug-related deaths were more common with megoestrol acetate. PMID- 11056334 TI - Faslodex (ICI 182780). an oestrogen receptor downregulator. AB - Anti-oestrogen therapy, tamoxifen in particular, has revolutionised the treatment of breast cancer. However, the partial agonist activity of tamoxifen is associated with an increased risk of endometrial cancer and the acquisition by patients of tamoxifen-resistance. In an attempt overcome these negative aspects of tamoxifen therapy, 'pure' anti-oestrogens have been developed and are currently being investigated for the treatment of breast cancer. PMID- 11056335 TI - Determination of tamoxifen and its metabolites in the endometrial tissue of long term treated women. AB - Concentrations of tamoxifen and its metabolites were analysed in the endometrium of 23 post-menopausal asymptomatic breast cancer patients who were on chronic tamoxifen therapy. Small endometrial samples were collected during diagnostic hysteroscopy. Analysis of both serum and tissue for these compounds was performed by mass spectrometry. Tamoxifen and its metabolites were far more concentrated in the endometrium than in serum; tamoxifen was also significantly more concentrated in endometrium with hyperplastic changes than in atrophic endometrium. Endometrial polyps of an additional 9 women showed a trend to a lesser concentration of compounds. Increased concentration of tamoxifen compounds could possibly be explained by the avidity of these compounds for oestrogen receptors (ER). PMID- 11056336 TI - Progesterone receptor activation. an alternative to SERMs in breast cancer. AB - Data regarding the effects of progesterone and a progestagen on human normal breast epithelial cell proliferation and apoptosis are presented here. In postmenopausal women, adding progesterone to percutaneously administrated oestradiol significantly reduces the proliferation induced by oestradiol. In vitro and in premenopausal women, stopping the administration of nomegestrol acetate triggers a peak of apoptosis. Fibro-adenoma and cancerous cells do not show this regulation of apoptosis. Progesterone seems to be important in normal breast homeostasis. PMID- 11056337 TI - Oestrogen, progesterone and androgen receptors in ovarian neoplasm. correlation between immunohistochemical and biochemical receptor analyses PMID- 11056338 TI - Tamoxifen modulates the apoptic pathway of primary endometrial cell culture PMID- 11056339 TI - The oestrogen receptor in epithelial ovarian carcinoma PMID- 11056340 TI - The prognostic significance of steroid receptor immunocytochemistry in endometrial carcinoma PMID- 11056341 TI - Oestrogen and progesterone receptors in ovarian cancer. correlation with clinico pathological features and activity of plasminogen activation system PMID- 11056342 TI - The effect of tamoxifen on endometrial thickness and endometrial histology PMID- 11056343 TI - Oestrogen differential interactions with ER+-responsive and ER--non-responsive human tumour cells PMID- 11056344 TI - Short term results of a computerised program for breast carcinoma risk analysis. PMID- 11056345 TI - Determination of tamoxifen and its metabolites in endometrial tissue of long-term treated women PMID- 11056346 TI - Downregulation of oestrogen receptor in advanced breast cancer after lipofection with wild-type (w-t) insulin growth factor binding protein IGFBP-2 cDNA plasmid PMID- 11056347 TI - Induction of PCD in tamoxifen-resistant oestrogen receptor positive (ER+) advanced breast cancer after combined therapy with ER antisense oligonucleotides and vinorelbine-tartrate encapsulated in DRV liposomes PMID- 11056348 TI - IUMPA - the modulation of the oestrogen receptor level of endometrial carcinoma by the intrauterine application of medroxyprogesterone acetate PMID- 11056349 TI - Impact of activation of MAP kinase family members on endocrine response and survival in clinical breast cancer PMID- 11056350 TI - Increased vascularity in norethisterone-based hormone replacement therapy compared with the natural cycle PMID- 11056351 TI - The prevalence of Ki 67 and oestrogen receptor beta antigens in the carotid arteries with atheromatous plaques compared with controls PMID- 11056352 TI - Oestrogen and progesterone receptor expression in postmenopausal endometrial polyps and their surrounding endometrium PMID- 11056353 TI - Heterogeneous oestrogen- and progesterone-receptor expression in low-grade endometrial stromal sarcomas-implications for therapy PMID- 11056354 TI - Oestrogen receptor is a critical component required for insulin-like growth factor (IGF)-mediated signalling and growth in MCF-7 cells PMID- 11056355 TI - A BRCA-negative breast and ovarian cancer lineage with unexplained reversible repetitive spontaneous abortion PMID- 11056356 TI - Endometrial monitoring in postmenopausal patients with breast cancer who are treated with tamoxifen. report Of 207 cases PMID- 11056357 TI - Inhibition of nitric oxide (NO) synthesis antagonises the oestrogen-induced increase in coronary blood PMID- 11056358 TI - Apoptosis and anti-apoptosis in oestrogen-receptor negative endometrial cancer cells in response to anastrozole, 4-hydroxytamoxifen and medroxyprogesterone acetate PMID- 11056359 TI - Third generation aromatase inhibitors and inactivators in the treatment and prevention of breast cancer PMID- 11056360 TI - Expression of oestrogen receptor alpha (ERalpha) and beta1 (ER-beta1) in endometrial adenocarcinoma PMID- 11056361 TI - The presence of (sub)endometrial cysts is not a suspicious sign in postmenopausal patients with breast cancer who are treated with tamoxifen PMID- 11056362 TI - Differences in oestrogen receptor alpha variant messenger RNAs between normal human breast tissue and primary breast carcinomas PMID- 11056363 TI - The modulation of oestrogen receptor-alpha activity by melatonin in MCF-7 human breast cancer cells PMID- 11056364 TI - Identification of a novel human oestrogen receptor (Delta receptor) and it's chromosomal localisation PMID- 11056365 TI - Radiotracer synthesis from [(11)C]-iodomethane: a remarkably simple captive solvent method. AB - A new method of [(11)C]-methylation is described, which attains the goals of simplicity, high radiochemical yields, speed, versatility, and automation. A standard high performance liquid chromatography (HPLC) injection loop on a standard HPLC injection valve is loaded with a solution (80 microL) of precursor (0.3-1.0 mg) in dimethyl formamide (DMF) or dimethyl sulfoxide (DMSO) (+ base if required). At ambient temperature [(11)C]-iodomethane is passed through the loop for 3-4 min with >90% trapping of activity. After a further 1-5 min, the contents of the loop are quantitatively injected onto the HPLC column for purification. Radiochemical yields are equal to or superior to conventional solution methods in all cases, even though no heat is applied. [(11)C]-labeled radiotracers that have been prepared by this method for human or animal studies include Raclopride, N methylspiperone, Ro 15-1788, FLB 457, RTI-32, Rolipram, SCH 23390, and SKF 82957. Since no vials, transfer lines, cooling, heating, or sealing valves are required, no transfer losses occur, yields are high, and cleanup is minimal, this "loop method" is ideal for most radiopharmaceuticals prepared from [(11)C]-iodomethane. PMID- 11056366 TI - (-)-N-[(11)C]propyl-norapomorphine: a positron-labeled dopamine agonist for PET imaging of D(2) receptors. AB - Imaging neuroreceptors with radiolabeled agonists might provide valuable information on the in vivo agonist affinity states of receptors of interest. We report here the radiosynthesis, biodistribution in rodents, and imaging studies in baboons of [(11)C]-labeled (-)-N-propyl-norapomorphine [(-)-NPA]. (-) [(11)C]NPA was prepared by reacting norapomorphine with [(11)C]propionyl chloride and a lithium aluminum hydride reduction. [(11)C]Propionyl chloride was prepared by reacting [(11)C]CO(2) with ethylmagnesium bromide, followed by reacting with phthaloyl chloride. The radiochemical yield of (-)-[(11)C]NPA was 2.5% at end of synthesis (EOS), and the synthesis time was 60 min. The specific activity was 1700+/-1900 mCi/micromol ( N=7; ranged 110-5200 mCi/micromol at EOS). Rodent biodistribution studies showed high uptake of [(11)C](-)-NPA in D(2) receptor rich areas, and the striatum/cerebellum ratios were 1.7, 3.4, and 4.4 at 5 min, 30 min, and 60 min postinjection, respectively. Pretreating the animals with haloperidol (1 mg/kg) decreased the striatum/cerebellum ratio at 30 min postinjection to 1.3. (-)-[(11)C]NPA was also evaluated via baboon positron emission tomography (PET) studies. Under control conditions ( N=4), rapid uptake of the tracer was observed and the striatum/cerebellum ratio reached 2.86+/-0.15 at 45 min postinjection. Following haloperidol pretreatment (0.2 mg/kg IV), the striatum/cerebellum ratio was 1.29 at 45 min postinjection. The result demonstrated the existence of specific binding of this new tracer to the D(2) receptor. To our knowledge, the current finding of a striatum/cerebellum ratio of 2.8 in baboon was the highest reported with a radiolabeled D(2) agonist. (-) [(11)C]NPA is a promising new D(2) agonist PET tracer for probing D(2) receptors in vivo using PET. PMID- 11056367 TI - Carbon-11-labeled KF21213: a highly selective ligand for mapping CNS adenosine A(2A) receptors with positron emission tomography. AB - In vivo assessment of the adenosine A(2A) receptors localized in the striatum with positron emission tomography (PET) may offers us a new diagnostic tool for neurological disorders. We evaluated the potential of [7-methyl-(11)C](E)-8-(2,3 dimethyl-4-methoxystyryl)-1, 3,7-trimethylxanthine ([(11)C]KF21213) as a PET ligand for mapping adenosine A(2A) receptors in the central nervous system. KF21213 showed a high affinity for the adenosine A(2A) receptors in vitro (Ki = 3.0 nM) and a very low affinity for the A(1) receptors (Ki > 10,000 nM). In mice, the striatal uptake of [(11)C]KF21213 increased for the first 15 min and then gradually decreased, whereas the uptake in the reference regions such as the cortex and cerebellum rapidly decreased. The uptake ratio of striatum to cortex and striatum to cerebellum increased to 8.6 and 10.5, respectively, at 60 min postinjection. The striatal uptake was significantly blocked by co-injection of carrier KF21213 or each of three other A(2A) antagonists, but not by co-injection of A(1) antagonist. The specific uptake was not detected in the cortex or in the cerebellum. Ex vivo autoradiography and PET clearly visualized adenosine A(2A) receptors in the rat striatum. [(11)C]KF21213 was the most selective tracer for mapping adenosine A(2A) in the central nervous system by PET among the tracers proposed to date. PMID- 11056368 TI - SPECT imaging with the D(4) receptor antagonist L-750,667 in nonhuman primate brain. AB - The suitability of an (123)I-labeled form of the putative D(4) receptor ligand L750,667 as a radiotracer for single photon emission computed tomography imaging was assessed in nonhuman primates. [(123)I]L750,667, labeled by iododestannylation, was administered to baboons in bolus and bolus plus constant infusion paradigms and imaged for 6 h. Total [(123)I]L750,667 brain uptake peaked (2.3% injected dose) at 15 min postinjection. [(123)I]L750,667 uptake was observed in all brain regions measured including diencephalon, brainstem, basal ganglia, cingulate cortex, and cerebellum, and slightly lower levels were noted in the frontal, parietal, temporoinsular, and occipital cortices. Administration of the D(4) receptor antagonist NGD 94-1 (2 mg/kg) did not displace radioactivity from any of the brain regions examined. Thus, while L750,667 is selective for the D(4) receptor in vitro, because brain [(123)I]L750,667 uptake was not displaced by NGD 94-1 at receptor saturating doses, [(123)I]L750,667 does not appear to be a suitable radiotracer for in vivo imaging of the D(4) receptor. PMID- 11056369 TI - Synthesis and binding characteristics of N-(1-naphthyl)-N'-(3-[(125)I] iodophenyl)-N'-methylguanidine ([(125)I]-CNS 1261): a potential SPECT agent for imaging NMDA receptor activation. AB - N-(1-Naphthyl)-N'-(3-[(125)I]-iodophenyl)-N'-methylguanidine ([(125)I]-CNS 1261) was synthesized as a potential radioligand to image N-methyl-D-aspartate (NMDA) receptor activation. [(125)I]-CNS 1261 was prepared by radioiodination of N-(1 naphthyl)-N'-(3-tributylstannylphenyl)-N'-methylguanidine using Na(125)I and peracetic acid. [(125)I]-CNS 1261 uptake in vivo reflected NMDA receptor distribution in normal rat brain, whereas in ischemic rat brain, uptake was markedly increased in areas of NMDA receptor activation. Radiolabeled CNS 1261 appears to be a good candidate for further development as a single photon emission computed tomography tracer in the investigation of NMDA receptor activation in cerebral ischemia. PMID- 11056370 TI - Synthesis and in vivo evaluation of [(11)C]CGP62349, a new GABA(B) receptor antagonist. AB - This paper describes the radiosynthesis of [(11)C]CGP62349, a potential ligand to assess GABA(B) receptors in vivo. (11)C was introduced by O-methylation of the corresponding des-methyl precursor, namely CGP67780. The final product was obtained with a reliable method in good yield. The radioligand was tested in monkey, revealing negligible blood-brain barrier penetration and brain uptake, thus prompting us to search for a new target structure with a better lipophilicity. PMID- 11056371 TI - Limitations in the use of low pH extraction to distinguish internalized from cell surface-bound radiolabeled antibody. AB - Internalization by cells of radiolabeled protein ligands bound to the cell surface is frequently analyzed by extraction of the cells with low pH buffers. This treatment supposedly strips the ligands from the cell surface, and remaining molecules are considered to be internalized. However, we show herein that: (1) low molecular weight catabolic products that are trapped within lysosomes (residualizing radiolabels) are efficiently extracted by low pH buffers, under the same conditions used to remove cell surface-bound material, and (2) low pH treatment lyses the majority of the cells, as shown with both a nonadherent and an adherent cell line, with the release of most of a (51)Cr label. Still, low pH extraction was effective at demonstrating Ab internalization, as has been demonstrated many times. These effects of low pH treatment may be attributed to the fixative properties of these buffers. Regardless of the mechanism, these data must be taken into consideration in interpreting the results of such experiments. PMID- 11056372 TI - Usefulness of (99m)Tc-d,l-HMPAO for estimation of GSH content in tumor tissues. AB - To investigate whether [(99m)Tc]-hexamethyl propyleneamine oxime ([(99m)Tc] HMPAO) is applicable for evaluating glutathione (GSH) localization in tumor, the difference of distribution between [(99m)Tc]-d,l- and meso-HMPAO was studied using a mouse tumor model. Biodistribution of [(99m)Tc]-d,l- or meso-HMPAO was studied in GSH-depleted and control Ehrlich tumor-bearing mice. GSH levels in tumors in GSH-depleted and control mice were measured in another set of mice. The uptake of [(99m)Tc]-d,l-HMPAO in tumor was significantly decreased by the diethyl maleate (DEM) treatment. On the other hand, the DEM treatment increased the accumulation of [(99m)Tc]-meso-HMPAO in tumor. Meanwhile, the content of GSH was lowest in tumor among the tissues tested and decreased in a manner similar to other tissues on preloading of DEM. [(99m)Tc]-d,l-HMPAO may be useful for estimating the GSH status in a certain tumor and thereby contribute to the diagnosis of anticancer therapy. PMID- 11056373 TI - Effect of chemotherapy on pulmonary epithelial permeability in lung cancer. AB - The aim of this study is to investigate the effect of one-course chemotherapy on the pulmonary epithelial permeability. Eighteen patients (18 male; mean age: 59+/ 10 years) with lung cancer (11 non-small cell, 7 small cell) inhaled 40 mCi (1,480 MBq) (99m)Tc-diethylenetriaminepentaacetic acid (DTPA). Thirty images of 1 min duration were acquired from posterior projection. The first 7 min of the decay-corrected time activity curves were used to calculate lung clearance half time. Clearance half-times of (99m)Tc-DTPA from the peripheral regions of the lungs were 42+/-19 min before and 56+/-34 min after chemotherapy (p=0.009); from the central regions, clearance half-times were 112+/-94 min before and 160+/-125 min after chemotherapy (p=0.005). This decrease in clearance rate might be related to decreasing mucociliary clearance rate due to the toxic effect of the chemotherapy regimen on cilia movement and/or mucus structure. (99m)Tc-DTPA radioaerosol study can be used to monitor the toxic effects of chemotherapy on the pulmonary epithelium and possibly on mucociliary function. PMID- 11056374 TI - Evaluation of (99m)Tc-labeled photosan-3, a hematoporphyrin derivative, as a potential radiopharmaceutical for tumor scintigraphy. AB - A quick and reproducible method for radiolabeling of Photosan-3(R), a photosensitizer used worldwide for photodynamic therapy (PDT) of cancer, with radioisotope of technetium ((99m)Tc) was developed. The radiotracer was evaluated for radiochemical purity, stability, and finally tissue distribution in a murine tumor model. The (99m)Tc-Photosan-3 prepared by using (99m)Tc-pertechnetate in place of reduced (99m)Tc demonstrated better labeling efficiency (>90%) and reproducibility. The procedure also minimized the radiation exposure to the radiochemist as handling time was considerably reduced. Due to the commercial availability of Photosan-3, the risk of batch-to-batch variation in the in situ synthesis of hematoporphyrin derivative, which is a complex mixture of at least five compounds, was also significantly reduced. The biodistribution studies and tumor scintigraphy confirmed that (99m)Tc-labeled Photosan-3 was preferentially taken up by the neoplastic tissue in a manner similar to the parent compound. In addition to applications in tumor imaging, (99m)Tc-Photosan-3 could also be used for estimating tumor uptake of Photosan-3 as may be required for individualization of clinical protocols of PDT. PMID- 11056375 TI - Evaluation in the baboon model of (99m)Tc-biguanide as a tracer for renal imaging. AB - Studies in various animal species have recently shown that (99m)Tc-BIG has practical and dosimetric benefits for renal imaging that could probably make it a good alternative to (99m)Tc-2, 3-dimercaptosuccinic acid ((99m)Tc-DMSA). In this study, using the baboon experimental model, the biodistribution of (99m)Tc-BIG and (99m)Tc-DMSA are compared. It is demonstrated that early good contrast imaging and more favourable dosimetry is possible with (99m)Tc-BIG compared to (99m)Tc-DMSA, confirming the quoted previous findings with small animals. Time activity curves for kidneys and other organs support these findings, and MIRDOSE software provided the dosimetry. PMID- 11056376 TI - Labeling proteins with Tc-99m via hydrazinonicotinamide (HYNIC): optimization of the conjugation reaction. AB - At present there is considerable interest in labeling peptides with Tc-99m for the development of target specific radiopharmaceuticals for imaging purposes. In the present study the conjugation of the bifunctional coupling agent succinimidyl hydrazinonicotinamide (S-HYNIC) was studied and optimized in a series of peptides [molecular weight (MW) 6.5-14.3 kDa]. Aprotinin (MW 6.5 kDa), cytochrome C (MW 12.4 kDa), alpha-lactalbumin (MW 14.2 kDa), and lysozyme (MW 14.3 kDa) were conjugated with S-HYNIC via the epsilon amino groups of their lysine residues. The effects of molar conjugation ratio, reaction temperature, pH, and protein concentration were studied. Reaction products were analyzed both with respect to the HYNIC-substitution ratio (spectrophotometrically) as well as to the labeling efficiency silica gel-instant thin layer chromatography (SG-ITLC) and molecular size fast performance liquid chromatography (FPLC). The effects of conjugation on biological activity were studied in three proteins binding to receptors on leukocytes: interleukin-8 (MW 8.5 kDa), interleukin-1alpha (MW 17 kDa), and interleukin-1 receptor antagonist (MW 17 kDa). The labeling efficiency of aprotinin, cytochrome c, alpha-lactalbumin, and lysozyme conjugated under optimal conjugation conditions exceeded 90%. Specific activities obtained were up to 7.5 MBq/microg. Conjugation was most efficient at 0 degrees C (as compared to 20 and 40 degrees C), at pH 8.2 (as compared to 6.0, 7.2, and 9.5), and at protein concentrations > or = 2. 5 mg/mL. In general, efficiency increased with increasing molar conjugation ratio (protein-HYNIC-ratio 1:3 < 1:6 < 1:15<1:30). For the receptor binding proteins, biological activity was preserved only under the mildest conjugation conditions. For each of these proteins an inverse relation between labeling efficiency and receptor binding capacity was found. Labeling proteins with (99m)Tc using S-HYNIC is easy, rapid, and efficient, and preparations with high specific activity can be obtained. However, biological activity of proteins may be lost at high HYNIC-substitution ratios. With the proteins tested here a careful balancing of reaction conditions resulted in acceptable, although suboptimal, labeling efficiencies and preservation of biological activity. PMID- 11056377 TI - Characterization of (111)In(3+) complexes of DTPA amide derivatives: biodistribution and clearance studied by gamma imaging. AB - A large series of structurally related diethylenetriaminepentaacetic acid amide derivatives with different structures and lipophilic properties were synthesized and radiolabeled with (111)In(3+). Two of the more hydrophobic compounds studied ([(111)In]L(9) and [(111)In]L(10)) showed high affinity for human serum albumin (HSA). The biodistribution and clearance properties shown by all complexes upon injection in Wistar rats were followed by gamma imaging. The blood retention time of the chelates correlates better with their binding to HSA than with their hydrophilic/lipophilic ratio. Hydrophilic and negatively charged complexes undergo renal retention, while the majority of the lipophilic complexes are retained in the blood for a longer period of time and are cleared through the liver. PMID- 11056378 TI - Biological energy from the igneous rock enhances cell growth and enzyme activity. AB - Some effects from natural resources might be ignored and unused by humans. Environmental hormesis could be a phenomena necessary to bio-organism existence on earth. Since 1919, radiation and some heavy metal hormesis from the environment were proved in various reports. In this study, igneous rock with very low radioactivity and high ferrous activity was measured by multichannel analyzer and inductively coupled plasma analyzer. The water treated by igneous rock, both directly soaked or indirectly in contact, induced increased activities of glucose oxidase, catalase, peroxidase, and superoxide dismutase. It also increased cell growth of SC-M1, HCT-15, Raji, and fibroblast cell lines. The water after treatment of igneous rock had no change in pH values, but displayed decreased conductivity values. We assume that the igneous rock could transfer energy to water to change the molecular structure or conformation of water cluster, or by radiation hormesis effect could then induce increased enzyme activity and cell growth. It is also possible that the energy from rock may combine radiation hormesis with other transferable biological energy forms to change water cluster conformation. PMID- 11056379 TI - The influence of tomograph sensitivity on kinetic parameter estimation in positron emission tomography imaging studies of the rat brain. AB - We investigated the influence of tomograph sensitivity on reliability of parameter estimation in positron emission tomography studies of the rat brain. The kinetics of two tracers in rat striatum and cerebellum were simulated. A typical injected dose of 10 MBq and a reduced dose of 1 MBq were assumed. Kinetic parameters were estimated using a region of interest (ROI) analysis and two pixel by-pixel analyses. Striatal binding potential was estimated as a function of effective tomograph sensitivity (S(eff)) using a simplified reference tissue model. A S(eff) value of > or =1% was required to ensure reliable parameter estimation for ROI analysis and a S(eff) of 3-6% was required for pixel-by-pixel analysis. We conclude that effective tomograph sensitivity of 3% may be an appropriate design goal for rat brain imaging. PMID- 11056380 TI - Column-switching HPLC for the analysis of plasma in PET imaging studies. AB - A column-switch high performance liquid chromatography method for the analysis of 4 mL of plasma is described with six examples of chromatography of [(11)C] labeled positron-emission tomography imaging agents. Complete extraction of all but the most polar metabolites by the reverse phase capture column is achieved by disruption of plasma protein binding by 8 M urea. PMID- 11056381 TI - Inhibitory effects of ML-9, wortmannin, and Y-27632 on the chemotaxis of vascular smooth muscle cells in response to platelet-derived growth factor-BB. AB - The chemotactic migration toward platelet-derived growth factor-BB of SM3, a cell line established from rabbit aorta smooth muscle, was examined by the Boyden chamber method. Myosin light-chain (MLC) kinase inhibitors ML-9 and wortmannin, and the Rho kinase inhibitor Y-27632 effectively reduced the migration. However, neither membrane ruffling nor the phosphorylation of MLC was inhibited concomitantly. The reduction is discussed with reference to a novel property of MLC kinase, which stimulates myosin ATPase activity without phosphorylating MLC [Ye et al. (1999) Proc. Natl. Acad. Sci. USA 96, 6666-6671]. PMID- 11056382 TI - Yeast Ulp1, an Smt3-specific protease, associates with nucleoporins. AB - Yeast Smt3 is a ubiquitin-like protein similar to the mammalian SUMO-1. Cdc3, a septin component, is known to be modified by Smt3. The level of this modification was affected by Smt3-specific protease mutation ulp1-ts or overexpression of ULP1. By two-hybrid screening, we isolated 5 UIP (Ulp1 interacting protein) genes. UIP1 was identical to NUP42 encoding a component of the nuclear pore complex (NPC). Gle1, another NPC-associating component, also interacted with Ulp1 in the two-hybrid system and co-immunoprecipitation experiment. Thus Ulp1 associates with nucleoporins and may interact with septin rings in the telophase. PMID- 11056383 TI - Differential scanning calorimetry study on the inner membrane lipids prepared from barotolerant Pseudomonas sp. BT1. AB - We investigated the properties of membrane lipids of barotolerant Pseudomonas sp. BT1 by differential scanning calorimetry and spectrophotometry using a system equipped with a hydrostatic pressure controller. In the case of cells grown under high pressure, an endothermic peak appeared under high-pressure measurement conditions. However, in the case of cells grown at 0.1 MPa, such a peak was not observed. It was also observed on spectrophotometry that the membrane lipids from cells grown at 30 MPa had stable properties in comparison with those grown at 0.1 MPa various hydrostatic pressures and temperatures. PMID- 11056384 TI - Stabilization of human recombinant erythropoietin through interactions with the highly branched N-glycans. AB - Human erythropoietin (EPO) produced in Chinese hamster ovary cells (CHO-EPO) is a hydrophobic protein stabilized by the highly branched complex-type N-glycans. To characterize the stabilizing effect of the N-glycans, the properties of enzymatically N-glycan-modified CHO-EPO species were compared spectrophotometrically. CD and fluorescence spectra following the protein unfolding induced by guanidine hydrochloride or pH revealed that the inner regions including the galactose residues of the N-glycans stabilize the protein conformation. The decrease in the conformational stability caused by enzymatic trimming of the N-glycans was associated with the exposure of the hydrophobic protein surface areas accessible to 1-anilino-8-naphthalenesulfonic acid (ANS) binding. Further, the ANS binding and heat denaturation of Escherichia coli expressed EPO (nonglycosylated EPO) were depressed in dilute solutions (1 mM or so) of free N-glycans of the complex type. These results, together with the finding that the N-glycans of CHO-EPO make little contact with the aromatic amino acid residues exposed on the protein surface, indicate that the inner regions including the galactose residues of the intramolecular N-glycans stabilize the protein conformation by clinging to the hydrophobic protein surface areas mainly made up of nonaromatic hydrocarbon groups. PMID- 11056385 TI - Double point mutant F34W/W140F of staphylococcal nuclease is in a molten globule state but highly competent to fold into a functional conformation. AB - The double point mutant F34W/W140F of staphylococcal nuclease was created and then characterized by far and near-UV CD, size-exclusion chromatography, ANS binding fluorescence. The results show that this mutant has properties consistent with the classical definition of a molten globule, i.e., substantial secondary structure but no unique tight packing of tertiary structure, a relatively compact size and a larger exposed hydrophobic surface area as compared with the wild type enzyme, indicating that a molten globule can occur under physiological conditions. However, the activity assay showed that the mutant still maintains wild-type levels of activity. To further clarify the mechanism of the substrate induced reactivation, enzymatic parameters such as K(M)(DNA), K(S)(DNA), K(M)(Ca), K(A)(Ca), K(d)(pdTp), and V(max) were determined, showing that all the parameters of this mutant are similar to those of the wild type enzyme. The results indicate that the F34W/W140F mutant has a similar substrate affinity to the wild type enzyme, and the functional conformation can be restored by substrate binding, which corresponds to the conformational adjustment capability of the mutant upon binding to ligands pdTp and Ca(2+). The severely disrupted tertiary structure and high activity of the mutant indicate that it is highly competent to fold to its functional conformation. The results suggest that the primary structure can only guide the mutant to a molten globule state and that ligand-binding causes the mutant to fold further into its functionally active conformation, indicating that ligand-binding plays an important role in protein folding and catalysis. PMID- 11056386 TI - Functional mapping against Escherichia coli for the broad-spectrum antimicrobial peptide, thanatin, based on an in vivo monitoring assay system. AB - Previously, we established for the first time an in vivo monitoring assay system conjugated with random mutagenesis in order to study the structure-function relationship of the antimicrobial peptide, apidaecin [Taguchi et al. (1996) Appl. Environ. Microbiol. 62, 4652-4655]. In the present study, this methodology was used to carry out the functional mapping of a second target, thanatin, a 21 residue peptide that exhibits the broadest antimicrobial spectrum so far observed among insect defense peptides [Fehlbaum et al. (1996) Proc. Natl. Acad. Sci. USA 93, 1221-1225]. First, a synthetic gene encoding thanatin was expressed in a fused form with Streptomyces protease inhibitor protein, SSI, under the control of tac promoter in Escherichia coli JM109. Expression of the thanatin-fused protein was found to depend on the concentration of the transcriptional inducer, isopropyl-beta-D-thio-galactopyranoside (IPTG), and to parallel the degree of growth inhibition of the transformant cells. When a PCR random mutation was introduced into the structural gene for thanatin, diminished growth inhibition of the IPTG-induced transformed cells was mostly observed in variants as measured by colony size (plate assay) or optical density (liquid assay) in comparison with the wild-type peptide, possibly depending on the decreased antimicrobial activity of each variant. Next, wild-type thanatin and three variants screened by the in vivo assay, two singly mutated proteins (C11Y and M21R) and one doubly mutated protein (K17R/R20G), were stably overproduced with a fusion partner protein resulting in the efficient formation of inclusion bodies in E. coli BL21(DE3). The products were isolated in large amounts (yield 30%) from the fused protein by successive chemical and enzymatic digestions at the protein fusion linker site. Anti-E. coli JM109 activities, judged by minimum inhibitory concentration, of the purified peptides were in good agreement with those estimated semi-quantitatively by the in vivo assay. Based on the NMR solution structure and molecular dynamics, the structure-function relationship of thanatin is discussed by comparing the functional mapping data obtained here with the previous biochemical data. The functional mapping newly suggests the importance of a hydrogen bonding network formed within the C-terminal loop joining the beta-strands arranged antiparallel to one another that are supposed to be crutial for exhibiting anti-E. coli activity. PMID- 11056387 TI - Characterization of recombinant human adipocyte-derived leucine aminopeptidase expressed in Chinese hamster ovary cells. AB - Adipocyte-derived leucine aminopeptidase (A-LAP) is a recently identified novel member of the M1 family of zinc-metallopeptidases. Transfection of the A-LAP cDNA into COS-7 cells resulted in the secretion of the enzyme. In this study, recombinant A-LAP was expressed in Chinese hamster ovary cells, purified to homogeneity and its enzymatic properties were characterized. The purified enzyme was active towards a synthetic substrate, L-leucyl-p-nitroanilide, yielding a V(max) of 3.55 micromol/min/mg and a K(m) of 1.28 mM, and was shown to be a monomeric protein with molecular mass of 120 kDa in solution. By monitoring the sequential N-terminal amino acid liberation, it was found that the enzyme hydrolyzes a variety of bioactive peptides, including angiotensin II and kallidin. Immunohistochemical analysis indicated that the enzyme is expressed in the cortex of the human kidney, where tissue kallikrein is localized. Taken together, these results indicate that A-LAP possesses a broad substrate specificity towards naturally occurring peptide hormones and suggest that it plays a role in the regulation of blood pressure through the inactivation of angiotensin II and/or the generation of bradykinin in the kidney. PMID- 11056388 TI - Molecular cloning, expression, and chromosomal mapping of human chondroitin 4 sulfotransferase, whose expression pattern in human tissues is different from that of chondroitin 6-sulfotransferase. AB - Chondroitin 4-sulfotransferase (C4ST) catalyzes the transfer of sulfate from 3' phosphoadenosine 5'-phosphosulfate to position 4 of the N-acetylgalactosamine residues of chondroitin. We previously reported the cloning of C4ST cDNA from mouse brain. We here report the cloning and expression of human C4ST cDNA. The cDNA was isolated from a human fetal brain cDNA library by hybridization with a DNA probe prepared from rat poly(A)(+) RNA used for the cloning of mouse C4ST cDNA. The cDNA comprises a single open reading frame that predicts a Type II transmembrane protein composed of 352 amino acids. The protein has an amino acid sequence homology of 96% with mouse C4ST. When the cDNA was introduced into a eukaryotic expression vector and transfected in COS-7 cells, the sulfotransferase activity that transfers sulfate to both chondroitin and desulfated dermatan sulfate was overexpressed. Northern blot analysis indicated that human C4ST mRNAs (6.0 and 1.9 kb) are expressed ubiquitously in various adult human tissues. Dot blot analysis has shown that human C4ST is strongly expressed in colorectal adenocarcinoma and peripheral blood leukocytes, whereas strong expression of human chondroitin 6-sulfotransferase (C6ST) is observed in aorta and testis. These observations suggest that the expression of C4ST and C6ST may be controlled differently in human tissues. The C4ST gene was localized to chromosome 12q23.2 q23.3 by fluorescence in situ hybridization. PMID- 11056389 TI - Aminoguanidine-treatment results in the inhibition of lens opacification and calpain-mediated proteolysis in Shumiya cataract rats (SCR). AB - The Shumiya cataract rat (SCR) is a hereditary cataract model in which lens opacity appears spontaneously in the nuclear and perinuclear portions at 11-12 weeks of age. We found incidentally that the oral administration of aminoguanidine (AG), an inhibitor of inducible nitric oxide synthase (iNOS), strongly inhibits the development of lens opacification in SCR. Since our previous results strongly suggested that calpain-mediated proteolysis contributes to lens opacification during cataract formation in SCR, we examined the calpain mediated proteolysis in AG-treated SCR lenses in detail. The results show that the calpain-mediated limited proteolysis of crystallins is also inhibited by AG treatment. However, the administration of AG has no effect on the substrate susceptibility to calpain. On the other hand, the autolytic activation of calpain in AG-treated lenses is strongly inhibited, although AG itself does not inhibit calpain activity in vitro. Then, we analyzed the effect of AG-treatment on calcium concentrations in lens, and found that the elevation in calcium concentration that should occur prior to cataractogenesis in lenses is strongly suppressed by AG-treatment. These results strengthen our previous conclusion that calpain-mediated proteolysis plays a critical role in the development of lens opacification in SCR. Moreover, our results indicate that the inhibition of calpain-mediated proteolysis by AG-treatment is due to the suppression of calcium ion influx into the lens cells. PMID- 11056390 TI - Relationships of p40(phox) with p67(phox) in the activation and expression of the human respiratory burst NADPH oxidase. AB - p40(phox) of the phagocyte NADPH oxidase forms a complex with p67(phox) in cytosol, and coincidentally decreases in patients who lack p67(phox). Here we investigated the mode of translocation of p40(phox) to the membrane, its cytoskeletal localization on activation of the NADPH oxidase, and the dependency of its expression relative to that of p67(phox). When human polymorphonuclear leukocytes (PMNs) were stimulated with phorbol myristate acetate (PMA), p40(phox) was translocated to the membrane along with p67(phox), and not was released into the cytosol. Studies with resting PMNs using Triton X-100 revealed the exclusive localization of p67(phox) in the cytoskeletal fraction. Unexpectedly, however, about half of p40(phox), which is deemed to be fully associated with p67(phox), was recovered in the non-cytoskeletal fraction. Unlike p47(phox), the association of p40(phox) with cytoskeleton was not induced by the PMA-stimulation. These results indicate not only that p40(phox) associates with cytoskeleton via a molecule of p67(phox), but also that there are distinct states of p40(phox) that can be manipulated with Triton X-100. Lastly, Western-blot analysis of hematopoietic cells revealed no correlation between p40(phox) and p67(phox) in their protein expressions during cell differentiation, and also that p40(phox) can be stably present alone in cells, unless in the case of mature PMNs. In this regard, definitive proof was obtained with Epstein-Barr virus-transformed B cells of a p67(phox)-deficient patient, in which p40(phox) was normally expressed. PMID- 11056391 TI - Effects of dimethyl sulfoxide, temperature, and sodium chloride on the activity of human matrix metalloproteinase 7 (matrilysin). AB - Effects of dimethyl sulfoxide (DMSO), temperature, and sodium chloride on the matrilysin-catalyzed hydrolysis of (7-methoxycoumarin-4-yl)acetyl-L-Pro-L-Leu-Gly L-Leu-[N(3)-(2, 4-dinitrophenyl)-L-2,3-diamino-propionyl]-L-Ala-L-Arg-NH(2) [MOCAc-PLGL(Dpa)AR] were examined. DMSO inhibited the matrilysin activity competitively with the inhibitor constant (K(i)) of 0. 59+/-0.04 M, and the binding between them was endothermic and entropy-driven. The binding of matrilysin with MOCAc-PLGL(Dpa)AR was also found to be entropy-driven. The matrilysin activity was increased in a biphasic exponential fashion with increasing concentration of NaCl, and was 5.3 times higher in the presence of 4 M NaCl than that in its absence. The first and second phases were separated at 0.5 M NaCl, and the activation at x M NaCl compared with the activity in the absence of NaCl was expressed as 2.1(x) at [NaCl] < 0.5 M and 1.4(x) at [NaCl] > 0.5 M. The activation was brought about solely through a decrease in the Michaelis constant (K(m)), and the catalytic constant (k(cat)) was not much altered. This suggests that the decrease in the electrostatic interaction and the increase in the hydrophobic interaction between matrilysin and the substrate might enhance the enzyme activity by reducing the K(m) value. PMID- 11056392 TI - Identification and characterization of novel isoforms of COP I subunits. AB - COP I-coated vesicles are involved in vesicular trafficking in the early secretory pathway. The COP I coat is composed of seven subunits, alpha-, beta-, beta'-, gamma-, delta-, epsilon-, and zeta-COPs. Evidence suggests, however, that there may be isoforms of the COP I subunits. In the present study, we identified homologs of gamma-COP (gamma2-COP; original gamma-COP is referred to as gamma1 COP in this paper) and of zeta-COP (zeta2-COP; original zeta-COP is referred to as zeta1-COP). gamma1- and gamma2-COPs, and zeta1- and zeta2-COPs share 80 and 75%, respectively, of amino acids. mRNAs for gamma2-COP and zeta2-COP are expressed ubiquitously, suggesting their fundamental role in cellular function. Immunofluorescence analysis shows that gamma2-COP and zeta2-COP are colocalized with beta-COP in the paranuclear cis-Golgi region. Yeast two-hybrid analysis indicates that gamma1- and gamma2-COPs can directly, albeit promiscuously, interact with zeta1- and zeta2-COPs. Like gamma1-COP, gamma2-COP can form a complex with beta-COP in vivo. The gamma1-COP-containing and gamma2-COP containing complexes can similarly interact with the cytoplasmic domain of p23. These results indicate that gamma2-COP and zeta2-COP can form a COP I-like complex in place of gamma1-COP and zeta1-COP, respectively, and suggest that the COP I complex and the COP I-like complex are functionally redundant. PMID- 11056393 TI - Functional role of Ca(2+)-binding site IV of scallop troponin C. AB - Scallop troponin C (TnC) binds only one Ca(2+)/mol and the single Ca(2+)-binding site has been suggested to be site IV on the basis of the primary structure [K. Nishita, H. Tanaka, and T. Ojima (1994) J. Biol. Chem. 269, 3464-3468; T. Ojima, H. Tanaka, and K. Nishita (1994) Arch. Biochem. Biophys. 311, 272-276]. In the present study, the functional role of Ca(2+)-binding site IV of akazara scallop (Chlamys nipponensis akazara) TnC in Ca(2+)-regulation was investigated using a site-directed mutant with an inactivated site IV (TnC-ZEQ), N- and C-terminal half molecule mutants (TnC(N) and TnC(C)), and wild-type TnC (TnC(W)). Equilibrium dialysis using (45)Ca(2+) demonstrated that TnC(W) and TnC(C) bind 0.6-0.8 mol of Ca(2+)/mol, but that TnC-ZEQ and TnC(N) bind virtually no Ca(2+). The UV difference spectra of TnC(W) and TnC(C) showed bands at around 280-290 nm due to the perturbation of Tyr and Trp upon Ca(2+)-binding, while TnC-ZEQ and TnC(N) did not show these bands. In addition, TnC(W) and TnC(C) showed retardation of elution from Sephacryl S-200 upon the addition of 1 mM CaCl(2), unlike TnC-ZEQ and TnC(N). These results indicate that Ca(2+) binds only to site IV and that Ca(2+)-binding causes structural changes in both the whole TnC molecule and the C-terminal half molecule. In addition, TnC(W), TnC-ZEQ, and TnC(C), but not TnC(N), were shown to form soluble complexes with scallop TnI at physiological ionic strength. On the other hand, the Mg-ATPase activity of reconstituted rabbit actomyosin in the presence of scallop tropomyosin was inhibited by scallop TnI and recovered by the addition of an equimolar amount of TnC(W), TnC-ZEQ, or TnC(C), but not TnC(N). These results imply that the site responsible for the association with TnI is located in the C-terminal half domain of TnC. Ternary complex constructed from scallop TnT, TnI, and TnC(W) conferred Ca(2+)-sensitivity to the Mg-ATPase of rabbit actomyosin to the same extent as native troponin, but the TnC(N)-TnT-TnI and TnC-ZEQ-TnT-TnI complexes conferred no Ca(2+)-sensitivity, while the TnC(C)-TnT-TnI complex conferred weak Ca(2+) sensitivity. Thus, the major functions of scallop TnC, such as Ca(2+)-binding and interaction with TnI, are located in the C-terminal domain, however, the full Ca(2+)-regulatory function requires the presence of the N-terminal domain. PMID- 11056394 TI - Molecular cloning and characterization of a new neuron-specific homologue of rat polypyrimidine tract binding protein. AB - Cancer-associated retinopathy (CAR) is a rare form of retinal degeneration and also one of the paraneoplastic neurologic disorders. Sera of CAR patients usually contain high titers of antibodies against retinal proteins, and CAR is believed to be an autoimmune disease. Using serum from a CAR patient as a molecular probe, a homologue of the polypyrimidine tract-binding protein (PTB) was isolated from a cDNA library of rat neonatal retina. This homologue, named PTB-like protein (PTBLP), encodes a 532 amino acid residue protein and has 73.5 and 68.8% homology with PTB and with a regulator of differentiation 1, respectively. Functional domains in the PTB, such as nuclear localization signals and four RNA recognition motifs (RRMs), were highly conserved. The expression of PTBLP mRNA was observed in the retina and brain but not in liver, kidney, spleen, or lung. The expression of PTBLP protein in rat retina was distributed in most of the cells in the ganglion cell layer and some cells in the inner nuclear layer. The PTBLP protein was localized in the nuclei of these cells. These results suggest that PTBLP is a new member of the PTB gene family and a neuron-specific homologue. PMID- 11056395 TI - An approach to the removal of yeast specific O-linked oligo-mannoses from human midkine expressed in Pichia pastoris using site-specific mutagenesis. AB - Human midkine is expressed and secreted in the medium under the control of an AOX1 gene promoter in Pichia pastoris using its own secretion signal. The midkine precursor is properly processed to yield the correct amino-terminus of mature midkine. However, more than half of the product receives yeast specific mannosylations. The sites for the mannosylations were determined to be the three threonine residues in the carboxy-terminal region of human midkine. In order to obtain non-mannosylated midkine, alanine residues were substituted for the three threonine residues by site specific mutagenesis. HPLC and mass spectrometry confirmed that the mutant midkine contained almost no mannose residues. Despite the amino acid substitutions in the carboxy-terminal region, mutant human midkine, promoted CHO cell proliferation as well as normal midkine. PMID- 11056396 TI - omega-hydroxylation activity toward leukotriene B(4) and polyunsaturated fatty acids in the human hepatoblastoma cell line, HepG2, and human lung adenocarcinoma cell line, A549. AB - The addition of glucose to the culture medium of HepG2 or A549 cells for 22 h caused a dose-dependent increase in leukotriene B(4) omega-hydroxylation activity in the homogenate. The addition of genistein to the culture medium of HepG2 or A549 cells for 22 h caused a dose-dependent decrease in the activity, although the number of living cells was not influenced by the addition of genistein. The inhibition by genistein was reversed by removal of genistein from the culture medium in 22 h. The specific leukotriene B(4) omega-hydroxylation activity was high in the nuclear envelope fraction of HepG2 or A549 cells, and a large portion of the activity was concentrated in the nuclear envelope fraction. In the nuclear envelope fraction, leukotriene B(4) omega-hydroxylation activity was accompanied by high polyunsaturated fatty acid omega-hydroxylation activity. The apparent K(m) values for arachidonic acid and leukotriene B(4) in the fractions of HepG2 or A549 cells were 25 and 50 microM, or 22 and 66 microM, respectively. The V(max) values were 222 and 104 pmol/min/mg protein, or 175 and 370 pmol/min/mg protein, respectively. NADPH-dependent omega-hydroxylation of LTB(4) in the nuclear envelope fraction of HepG2 or A549 cells was strongly inhibited by metyrapone and CO. The expression of cytochrome P450 4F2 mRNAs was detected in HepG2 and A549 cells, and thus the arachidonic acid and leukotriene B(4) omega hydroxylation activities in the nuclear envelope fractions of HepG2 and A549 cells are likely due to cytochrome P450 4F2. PMID- 11056397 TI - Cholesterol homeostasis in rat astrocytoma cells GA-1. AB - Astrocytes play a key role in cholesterol metabolism in central nervous system. We have shown that fetal rat astrocytes in primary culture secrete cholesterol rich HDL with the endogenous apolipoprotein (apo) E and generate cholesterol-poor HDL with exogenous apoE and apoA-I [Ito et al. (1999) J. Neurochem. 72, 2362]. In order to study these reactions in relation to the stage of cell differentiation, we examined generation of HDL by rat astrocytoma cells. Lack of apoE secretion was found in three astrocytoma cell lines, human T98G, rat C6, and GA-1 [Kano Tanaka et al. (1986) Proc. Jpn. Acad. Ser. B 62, 109]. GA-1 produced apoE at very low level and therefore generated much less HDL by itself than the astrocytes in primary culture. In contrast, GA-1 interacted with exogenous apoE and apoA-I to produce cholesterol-rich HDL while the astrocytes produced cholesterol-poor HDL with these apolipoproteins. Cholesterol biosynthesis rate measured from mevalonate was higher and down-regulated more by LDL in the astrocytes than GA-1. On the other hand, the cellular cholesterol level, uptake of LDL, and cyclodextrin-mediated non-specific diffusion of cholesterol from cell surface were same between these two cells. Treatment of GA-1 with acidic fibroblast growth factor influenced neither the production of apoE nor the baseline lipid secretion, but increased the cholesterol synthesis from mevalonate and the magnitude of its down-regulation by LDL, and decreased cholesterol content in the HDL produced by exogenous apoA-I. In conclusion, suppression of apoE biosynthesis in the undifferentiated astrocytes GA-1 resulted in poor secretion of cholesterol rich HDL and in turn more production of HDL with exogenous apolipoprotein. Cellular cholesterol homeostasis was altered accordingly. PMID- 11056398 TI - A novel imprinted gene, KCNQ1DN, within the WT2 critical region of human chromosome 11p15.5 and its reduced expression in Wilms' tumors. AB - WT2 is defined by a maternal-specific loss of heterozygosity on human chromosome 11p15.5 in Wilms' and other embryonal tumors. Therefore, the imprinted genes in this region are candidates for involvement in Wilms' tumorigenesis. We now report a novel imprinted gene, KCNQ1DN (KCNQ1 downstream neighbor). This gene is located between p57(KIP2) and KvLQT1 (KCNQ1) of 11p15.5 within the WT2 critical region. KCNQ1DN is imprinted and expressed from the maternal allele. We examined the expression of KCNQ1DN in Wilms' tumors. Seven of eighteen (39%) samples showed no expression. In contrast, other maternal imprinted genes in this region, including p57(KIP2), IMPT1, and IPL exhibited almost normal expression in these samples, although some samples expressed IGF2 biallelically. These results suggest that KCNQ1DN existing far from the H19/IGF2 region may play some role in Wilms' tumorigenesis along with IGF2. PMID- 11056399 TI - Isolectins from Solanum tuberosum with different detailed carbohydrate binding specificities: unexpected recognition of lactosylceramide by N-acetyllactosamine binding lectins. AB - Glycosphingolipid recognition by two isolectins from Solanum tuberosum was compared by the chromatogram binding assay. One lectin (PL-I) was isolated from potato tubers by affinity chromatography, and identified by MALDI-TOF mass spectrometry as a homodimer with a subunit molecular mass of 63,000. The other (PL-II) was a commercial lectin, characterized as two homodimeric isolectins with subunit molecular masses of 52,000 and 55,000, respectively. Both lectins recognized N-acetyllactosamine-containing glycosphingolipids, but the fine details of their carbohydrate binding specificities differed. PL-II preferentially bound to glycosphingolipids with N-acetyllactosamine branches, as Galbeta4GlcNAcbeta6(Galbeta4GlcNAcbeta3)Galbeta4Glcbeta1C er. PL-I also recognized this glycosphingolipid, but bound equally well to the linear glycosphingolipid Galbeta4GlcNAcbeta3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer. Neolactotetraosylceramide and the B5 pentaglycosylceramide were also bound by PL I, while other glycosphingolipids with only one N-acetyllactosamine unit were non binding. Surprisingly, both lectins also bound to lactosylceramide, with an absolute requirement for sphingosine and non-hydroxy fatty acids. The inhibition of binding to both lactosylceramide and N-acetyllactosamine-containing glycosphingolipids by N-acetylchitotetraose suggests that lactosylceramide is also accomodated within the N-acetylchitotetraose/N-acetyllactosamine-binding sites of the lectins. Through docking of glycosphingolipids onto a three dimensional model of the PL-I hevein binding domain, a Galbeta4GlcNAcbeta3Galbeta4 binding epitope was defined. Furthermore, direct involvement of the ceramide in the binding of lactosylceramide was suggested. PMID- 11056400 TI - Efficient cloning and engineering of giant DNAs in a novel Bacillus subtilis genome vector. AB - The Genome of Bacillus subtilis 168 was used for cloning and engineering of large sized DNAs. A mouse genomic DNA of approximately 120 kb was cloned into a locus of the B. subtilis genome by ordered assembly of 20- to 50-kb mouse DNA segments. Cloned mouse DNA, maintained stably, was engineered through B. subtilis transformation and recombination. Creation of an I-PpoI recognition sequence at both ends of the insert facilitated its isolation by pulsed field gel electrophoresis. The basic concept of genome vector technology is suited to the handling of DNAs larger than 100 kb. PMID- 11056401 TI - Design and synthesis of sensitive fluorogenic substrates specific for Lys gingipain. AB - Lys-gingipain (Kgp) is a major cysteine proteinase produced by the oral anaerobic bacterium Porphyromonas gingivalis, and has been implicated as a major pathogen in the development and progression of advanced adult periodontitis. This enzyme is believed to act as a major virulence factor of the disease, yet there exist no convenient and sensitive substrates for analyzing its biological activity. For a better understanding of the importance of this enzyme in the organism, there is an urgent need for specific substrates. Here we designed and synthesized two peptide 4-methyl-coumaryl-7-amides (MCA), carbobenzoxy (Z)-His-Glu-Lys-MCA, and Z Glu-Lys-MCA, and tested their possible use as sensitive substrates for Kgp with limited specificity. Both substrates exhibited greater k(cat)/K(m) values than the best known Kgp substrates described so far. Both substrates were resistant to Arg-gingipain, another pathogenic cysteine proteinase from P. gingivalis, as well as trypsin and cathepsins B, L, and H. The levels of Kgp in various microorganisms and human cells were determined with Z-His-Glu-Lys-MCA. Little or no Kgp-like activity was detected in either other microorganisms or human cells tested. These results indicate that the present substrates are a valuable and fast tool for routine assays and for mechanistic studies on Kgp. PMID- 11056402 TI - Polynucleotide:Adenosine glycosidase is the sole activity of ribosome inactivating proteins on DNA. AB - Polynucleotide: adenosine glycosidases (PNAG) are a class of plant and bacterial enzymes commonly known as ribosome-inactivating proteins (RIP). They are presently classified as rRNA N-glycosidases in the enzyme nomenclature [EC 3.2.2.22]. Several activities on nucleic acids, other than depurination, have been attributed to PNAG: in particular modifications induced in circular plasmids, including linearisation and topological changes, and cleavage of guanidinic residues. Here we describe a chromatographic procedure to obtain nuclease-free PNAG by dye-chromatography onto Procion Red derivatized Sepharose((R)). Highly purified enzymes depurinate extensively pBR322 circular, supercoiled DNA at neutral pH and exhibit neither DNase nor DNA glycolyase activities, do not cause topological changes, and adenine is the only base released from DNA and rRNA, even at very high enzyme concentrations. A scanning force microscopy (SFM) study of pBR322 treated with saporin-S6 confirmed that (i) this PNAG binds extensively to the plasmid, (ii) the distribution of the bound saporin-S6 molecules along the DNA chain is markedly variable, (iii) plasmids already digested with saporin-S6 do not appear fragmented or topologically modified. The observations here described demonstrate that polynucleotide:adenosine glycosidase is the sole enzymatic activity of the four ribosome-inactivating proteins gelonin, momordin I, pokeweed antiviral protein from seeds and saporin-S6. These proteins belong to different families, suggesting that the findings here described may be generalized to all PNAG. PMID- 11056403 TI - Increasing community participation after brain injury: strategies for identifying and reducing the risks. AB - This article describes the complex processes involved in evaluating safety, judgment, and risk after brain injury. Starting with a review of common risk factors after onset, the article then moves to a discussion of the dilemmas faced by family members and clinicians alike when determining the individual's level of risk. Numerous suggestions are offered to identify support system concerns and to establish specific barriers to independence. The need to adopt a systematized approach to empirically verifying concerns about safety is emphasized, in addition to utilizing therapeutic interventions that are sensitive to the individual's learning capacity, psychological status, and environmental influences. The article concludes with examples of how to structure support for individuals who present with ongoing risk factors but still need to experience greater levels of personal freedom. PMID- 11056404 TI - Service utilization following traumatic brain injury. AB - OBJECTIVES: To document service utilization by people with a traumatic brain injury at different times postinjury and to identify factors that predict service use. DESIGN: Cross-sectional study design. Four groups of subjects were randomly selected from a regional database, according to their time postinjury: 6-18 months; 2-4 years; 6-9 years; and 10-17 years. SUBJECTS: A total of 119 adults with a traumatic brain injury (TBI). SETTING: Hospital and community-based clients in Sydney, Australia. OUTCOME MEASURES: Glasgow Outcome Scale, Disability Rating Scale; Functional Independence Measure; Lidcombe Psychosocial Disability Scale; number, type, and frequency of services used in the previous 12 months. RESULTS: Subjects in all four groups used a variety of services. The mean number of services used was 4.2, and there was only a moderate decline in service use over time. The use of medical and allied health services remained high in all four groups. Severity of injury, physical and cognitive disability, and psychosocial disability were all predictors of service utilization. Psychosocial disability was strongly associated with ongoing service utilization. CONCLUSION: In this study, people with TBI used services well beyond the early stage of recovery. Psychosocial disability may be a better predictor of service use than physical and cognitive disability alone. PMID- 11056405 TI - Intellectual and emotional functioning in college students following mild traumatic brain injury in childhood and adolescence. AB - OBJECTIVE: To investigate whether college students with a history of mild traumatic brain injury (TBI) in childhood or adolescence show residual deficits in intellectual functioning, approaches to studying, or emotional stability. DESIGN: Participants with a history of mild TBI and two control groups. SETTING: Volunteers were recruited from students taking an introductory psychology course. PARTICIPANTS: 79 students with a history of mild TBI, 75 students with a history of general anesthesia, and 93 students with no history of either TBI or general anesthesia. MAIN OUTCOME MEASURES: Participants carried out tests of verbal memory, nonverbal memory, verbal fluency, and nonverbal fluency; in addition, they completed a short form of the Approaches to Studying Inventory and the Symptom Checklist-90-Revised (SCL-90-R). RESULTS: In comparison with the two control groups, the students with a history of mild TBI produced similar scores on the cognitive tests and similar orientations to studying. However, they showed a significantly higher level of emotional distress on the SCL-90-R. CONCLUSION: College students with a history of mild TBI in childhood or adolescence are intellectually unimpaired and approach their studying in a similar manner to their uninjured classmates. Nevertheless, they report more severe distress in terms of their general personal and emotional functioning. PMID- 11056406 TI - Affective disorders after traumatic brain injury: cautions in the use of the Symptom Checklist-90-R. AB - DESIGN: The current study compared the Symptom Checklist 90-Revised (SCL-90-R) results from a population of 88 individuals with traumatic brain injury (TBI) to those of a normative group. SETTING: Individuals with traumatic brain injury referred to a university-based psychology clinic. RESULTS: Results corroborated those from a smaller study,(1) where a considerable proportion of the elevations found on the subscales was attributed to endorsement of items identified as having a neurological basis. CONCLUSION: This reinforces the view that the results of neurological groups on clinical measures of emotional distress that have been standardized on physically healthy populations can be distorted because many of the items can be answered from physical and emotional perspectives. PMID- 11056407 TI - Coping strategies and emotional outcome following traumatic brain injury: a comparison with orthopedic patients. AB - OBJECTIVES: To investigate coping strategies in relation to emotional adjustment in individuals with traumatic brain injury (TBI) 1-5 years postinjury and to compare these with a group of 40 participants who sustained serious orthopedic injuries. DESIGN: Participants completed measures of handicap and coping strategies, and rated their levels of depression, anxiety, and self-esteem on standardized questionnaires. SETTING: Participants had received inpatient rehabilitation at Bethesda Hospital 1-5 years prior to completing questionnaires. They were recruited from a list of consecutive admissions. PARTICIPANTS: 88 TBI individuals were compared with 40 participants who had sustained serious orthopedic injuries without damage to the central nervous system. They had all been involved in motor vehicle or work-related accidents. OUTCOME MEASURES: Beck Depression Inventory (BDI) and State-Trait Anxiety Inventory (STAI). RESULTS: Consistent with previous studies; a significant proportion of the current sample displayed high levels of emotional distress. Results showed few differences between the TBI and orthopedic groups. Coping strategies characterized by worry, wishful thinking, and self-blame were associated with higher levels of depression and anxiety in both groups. Strategies focusing on problem solving and having a positive outlook were related to lower anxiety levels, but to a lesser degree. CONCLUSIONS: This study has provided further evidence that coping strategies are associated with emotional outcome in TBI individuals. There is now a growing empirical basis on which preliminary interventions can be based. PMID- 11056408 TI - Developing a suicide prevention strategy based on the perspectives of people with brain injuries. AB - OBJECTIVE: To develop a service-relevant, suicide prevention strategy based on the perspectives of people with brain injuries and their family members. DESIGN: Structured interview-based, qualitative case design. SETTING: Interviews were conducted in the context of community-based brain injury rehabilitation service delivery. PARTICIPANTS: Ten persons with moderate to severe brain injuries who exhibited suicidal orientations and four family member/carers of these participants. RESULTS: Qualitative analysis of interview transcripts revealed a number of relevant themes. The primary theme was that informal relationships play a key role in preventing suicide. Secondary themes included the potential role of specialist brain injury rehabilitation services in suicide prevention and the need for provision of more information about brain injury to family and friends to promote understanding. CONCLUSIONS: Some discrepancy was noted between the perspectives of people with brain injuries and family members. The need for multiple strategies to respond to suicide risk was reinforced. Service-relevant resources (suicide risk screen, contract, and brochure) have been developed and included in service delivery. PMID- 11056409 TI - Improvement/rehabilitation of memory functioning with neurotherapy/QEEG biofeedback. AB - This article presents a new approach to the remediation of memory deficits by studying the electrophysiological functioning involved in memory and applying biofeedback techniques. A Quantitative EEG (QEEG) activation database was obtained with 59 right-handed subjects during two auditory memory tasks (prose passages and word lists). Memory performance was correlated with the QEEG variables. Clinical cases were administered the same QEEG activation study to determine their deviations from the values that predicted success for the reference group. EEG biofeedback interventions were designed to increase the value (to normal levels) of the specific electrophysiological variable that was related to successful memory function and deviant in the subject. Case examples are presented that indicate the successful use of this intervention style in normal subjects and in subjects with brain injury; improvement cannot be fully explained by spontaneous recovery, given the time postinjury. Five cases (two normal, two subjects with brain injury, and one subject who had stereotactic surgery of the hippocampus for seizure control) are presented. Improvements ranged from 68% to 181% in the group of patients with brain injury, as a result of the interventions. PMID- 11056410 TI - ADHD, Ritalin, and big brother. PMID- 11056411 TI - Attention deficit/hyperactivity disorder (ADHD) in children: rationale for its integrative management. AB - Attention Deficit/Hyperactivity Disorder (ADHD) is the most common behavioral disorder in children. ADHD is characterized by attention deficit, impulsivity, and sometimes overactivity ("hyperactivity"). The diagnosis is empirical, with no objective confirmation available to date from laboratory measures. ADHD begins in childhood and often persists into adulthood. The exact etiology is unknown; genetics plays a role, but major etiologic contributors also include adverse responses to food additives, intolerances to foods, sensitivities to environmental chemicals, molds, and fungi, and exposures to neurodevelopmental toxins such as heavy metals and organohalide pollutants. Thyroid hypofunction may be a common denominator linking toxic insults with ADHD symptomatologies. Abnormalities in the frontostriatal brain circuitry and possible hypofunctioning of dopaminergic pathways are apparent in ADHD, and are consistent with the benefits obtained in some instances by the use of methylphenidate (Ritalin) and other potent psychostimulants. Mounting controversy over the widespread use of methylphenidate and possible life-threatening effects from its long-term use make it imperative that alternative modalities be implemented for ADHD management. Nutrient deficiencies are common in ADHD; supplementation with minerals, the B vitamins (added in singly), omega-3 and omega-6 essential fatty acids, flavonoids, and the essential phospholipid phosphatidylserine (PS) can ameliorate ADHD symptoms. When individually managed with supplementation, dietary modification, detoxification, correction of intestinal dysbiosis, and other features of a wholistic/integrative program of management, the ADHD subject can lead a normal and productive life. PMID- 11056412 TI - The use of nebulized glutathione in the treatment of emphysema: a case report. AB - We present the case of a 95-year-old man with an acute respiratory crisis secondary to emphysema and apparent bronchial infection. Treatment with nebulized glutathione led to a rapid resolution of the crisis, as well as a marked improvement in the chronic course of the disease. This treatment has been used since for a number of patients with emphysema. The safety and bioavailability of this method of delivery have been established in human studies. Preliminary results suggest efficacy for nebulized administration of glutathione in this patient population. We suggest this treatment can be considered an option for acute respiratory crises due to COPD. PMID- 11056413 TI - Environmental medicine, part 4: pesticides - biologically persistent and ubiquitous toxins. AB - Although the use of pesticides has doubled every ten years since 1945, pest damage to crops is more prevalent now than it was then. Many pests are now pesticide resistant due to the ubiquitous presence of pesticides in our environment. Chlorinated pesticide residues are present in the air, soil, and water, with a concomitant presence in humans. Organophosphate and carbamate pesticides - the compounds comprising the bulk of current pesticide use - are carried around the globe on air currents. Municipalities, schools, churches, business offices, apartment buildings, grocery stores, and homeowners use pesticides on a regular basis. Pesticides are neurotoxins that can cause acute symptoms as well as chronic effects from repeated low-dose exposure. These compounds can also adversely affect the immune system, causing cell-mediated immune deficiency, allergy, and autoimmune states. Certain cancers are also associated with pesticide exposure. Multiple endocrine effects, which can alter reproduction and stress-handling capacity, can also be found. Limited testing is available to assess the toxic overload of these compounds, including serum pesticide levels and immune system parameters. Treatment for acute or chronic effects of these toxins includes avoidance, supplementation, and possibly cleansing. PMID- 11056414 TI - Natural treatment of perennial allergic rhinitis. AB - Perennial allergic rhinitis is an IgE-mediated inflammatory disorder of the nasal mucosa characterized by paroxysms of sneezing, nasal congestion, pruritus, and rhinorrhea. The condition may be caused by certain environmental agents, food sensitivities, structural abnormalities, metabolic conditions, or synthetic drugs. Recent health impairment outcome studies on allergic rhinitis sufferers reveal a measurable decline in physical and mental health status and the inability to perform daily activities. Antihistamines, decongestants, anticholinergic agents, and corticosteroid drug therapy, alone or in combination, are typically used in the treatment of allergic rhinitis. Reported adverse side effects include sedation, impaired learning/memory, and cardiac arrhythmias. Therapeutic strategies should seek to decrease the morbidity already associated with this condition. Urtica dioica, bromelain, quercetin, N-acetylcysteine, and vitamin C are safe, natural therapies that may be used as primary therapy or in conjunction with conventional methods. PMID- 11056415 TI - The use of ascorbigen in the treatment of fibromyalgia patients: a preliminary trial. AB - Twelve female fibromyalgia syndrome (FMS) patients were given 500 mg per day of a blend containing 100 mg ascorbigen and 400 mg broccoli powder in a preliminary, one-month, open-label trial. This group of patients showed a mean 20.1 percent (p=0.044) decrease in their physical impairment score and a mean 17.8 percent (p=0.016) decrease in their total fibromyalgia impact scores as measured by the Fibromyalgia Impact Questionnaire. The mean physical impairment score two weeks post-treatment showed a significant return to near pre-treatment level (p=0.028). Analysis of ten of the patients' mean threshold pain levels at the 18 possible tender points obtained before and at the end of treatment showed a strong trend toward an increase in the mean threshold pain level (p=0.059). The reduced sensitivity to pain and improvement in quality of life measured in this study appear to be clinically relevant and a larger, double-blind study is warranted. PMID- 11056416 TI - Larch arabinogalactan. AB - Larch arabinogalactan is a polysaccharide powder derived from the wood of the larch tree (Larix species) and comprised of approximately 98 percent arabinogalactan. Arabinogalactans are found in a variety of plants but are more abundant in the Larix genus, primarily Larix occidentalis (Western Larch). The Western Larch is unique among pines in that it loses its needles in the fall. Western Larch is also known as Mountain Larch or Western Tamarack and is native to the Pacific and Inland Northwest United States as well as parts of British Columbia, Canada. Larch arabinogalactan is approved by the U.S. Food and Drug Administration (FDA) as a source of dietary fiber, but also has potential therapeutic benefits as an immune stimulating agent and cancer protocol adjunct. PMID- 11056417 TI - N-acetylcysteine. AB - N-acetylcysteine (NAC) is the acetylated precursor of both the amino acid L cysteine and reduced glutathione (GSH). Historically it has been used as a mucolytic agent in chronic respiratory illnesses as well as an antidote for hepatotoxicity due to acetaminophen overdose. More recently, animal and human studies of NAC have shown it to be a powerful antioxidant and a potential therapeutic agent in the treatment of cancer, heart disease, HIV infection, heavy metal toxicity, and other diseases characterized by free radical oxidant damage. NAC has also been shown to be of some value in treating Sjogren's syndrome, smoking cessation, influenza, hepatitis C, and myoclonus epilepsy. PMID- 11056418 TI - The EGF receptor - an essential regulator of multiple epidermal functions. AB - Epidermal keratinocytes express both the epidermal growth factor receptor (EGFR) and several of its ligands, establishing the constitutive elements of an autocrine loop in this cell type. Activation of the EGFR provides signals essential to several aspects of normal keratinocyte biology including cell cycle progression, differentiation, cell movement and cellular survival. It may be argued that enhanced keratinocyte survival via EGFR activation is the most important function as it limits the manifestation of other phenotypes. The frequent deregulation of EGFR expression and activation in benign and malignant hyperproliferative skin diseases motivates the investigation of EGFR-dependent intracellular pathways which contribute to the varied EGFR-dependent phenotypes. PMID- 11056419 TI - Phylloid hypomelanosis is closely related to mosaic trisomy 13. AB - Phylloid hypomelanosis is a distinct type of pigmentary mosaicism characterized by congenital hypochromic macules resembling a floral ornament with various elements such as round or oval patches, macules resembling the asymmetrical leaves of a begonia, or oblong lesions. A review of cases with documentation of cytogenetic findings showed that aberrations involving chromosome 13 were present in 5 out of 6 patients. Examination of blood lymphocytes revealed a 46, XX/47 XX, +13 or 46, XX/47, XX, +der (13) mosaic in three of these cases and a karyotype 46, XX, t(13;13) in the other two cases. Cytogenetic analysis of skin fibroblasts showed chromosomal mosaicism in 4 of the 5 patients. In the remaining case, a chromosome 13 translocated on 13 was found in 100% of blood lymphocytes and skin fibroblasts, suggesting that mosaicism involving chromosome 13 may have developed in the melanocyte system. In conclusion, contrasting with hypomelanosis of Ito which is a cutaneous sign of many different states of mosaicism, phylloid hypomelanosis seems to originate preponderantly from a mosaic state involving chromosome 13. Future case reports may help to delineate further the significance of this relationship. PMID- 11056420 TI - Preclinical diagnosis of pseudoxanthoma elasticum - methodological restrictions and ethical problems. AB - Pseudoxanthoma elasticum (PXE) is an inherited connective tissue disease. Only recently, mutations in the MRP6 gene on chromosome 16p13.1 have been identified in PXE families. Up to now, predictive testing has not been available. Since ultrastructural connective tissue alterations in overtly normal skin of predilection sites have supported preclinical diagnosis in children of affected individuals, we have screened the daughters of a PXE patient for these alterations. The patient's biopsy from lesional skin revealed elastin and collagen fibril abnormalities, but biopsies from the clinically inconspicuous daughters showed only ultrastructural alterations of collagen fibrils. These findings are inconclusive regarding the diagnosis of PXE in the daughters. Predictive or preclinical diagnosis of incurable, late-onset disorders creates complex social, ethical, and legal problems which call for special management strategies. PMID- 11056421 TI - The treatment of psoriasis with etretinate and acitretin: a follow up of actual use. AB - The present study is a retrospective search on the actual use of systemic retinoids in the treatment of psoriasis. The design of the study was inclusion of the patients in whom retinoid treatment was initiated from 1981 up to 1989. The analysis was carried out in 1999, after at least 10 years of follow up. In total 94 patients were included, who were treated with etretinate or acitretin, out of a cohort of 2,000 patients with psoriasis at the Nijmegen Department of Dermatology. The majority of the patients were older than 40 years, 31% were suffering from pustular psoriasis, 6% from erythrodermic psoriasis and 17% from psoriasis arthropatica. Most patients had had psoriasis for more than 5 years and in 56% of them retinoids were the first systemic treatment. Continuous treatment for more than one year was recorded in 33% of the patients. During long-term follow up of at least 10 years, 25% of them were included again for acitretin treatment. Therefore, prolonged treatment actually had occurred in approximately half of the patients. In contrast to the common belief, erythrodermic psoriasis proved to be not the typically "low-dose" - and pustular psoriasis was not the typically "high dose" indication. The present study, however, reconfirmed the high efficacy of systemic retinoids in pustular- and erythrodermic psoriasis. It was also reconfirmed that systemic retinoids are not effective in arthropathic psoriasis. The occurrence of side effects largely followed the controlled investigations. At least 10 years' follow up had not revealed serious side effects. In those patients (n = 30) who completed the entire survey in the Nijmegen centre 19 patients were treated with at least one course of photo(chemo)therapy and 9 patients were treated subsequently with methotrexate. PMID- 11056422 TI - Upregulation of adhesion complex proteins and fibronectin by human keratinocytes treated with an aqueous extract from the leaves of Chromolaena odorata (Eupolin). AB - The fresh leaves and extract of the plant Chromolaena odorata are a traditional herbal treatment in developing countries for burns, soft tissue wounds and skin infections. We have previously shown that the extract had an effect on the growth and proliferation of keratinocytes and fibroblasts in culture. This study has demonstrated that Eupolin extract increased expression of several components of the adhesion complex and fibronectin by human keratinocytes. Using indirect immunofluorescence we found increased expression (dose-dependent) of laminin 5, laminin 1, collagen IV, and fibronectin. The expression of the b1 and b4 integrins was upregulated by the extract at low concentrations (0.1 and 1 microg/ml), but the expression was decreased at higher doses of Eupolin (10 microg-150 microg/ml). A number of clinical studies carried out by Vietnamese and international medical investigators have demonstrated the efficacy of this extract on the wound healing process. In this study we have shown that Eupolin stimulated the expression of many proteins of the adhesion complex and fibronectin by human keratinocytes. The adhesion complex proteins are essential to stabilise epithelium and this effect could contribute to the clinical efficacy of Eupolin in healing. PMID- 11056423 TI - Four cases of sebopsoriasis or seborrheic dermatitis of the face and scalp successfully treated with 1a-24 (R)-dihydroxycholecalciferol (tacalcitol) cream. AB - A 71-year-old woman visited our clinic due to the presence of widespread scaly erythema on her face, scalp, and lower extremities. She was tentatively diagnosed as having seborrheic dermatitis but the symptoms were difficult to distinguish from psoriasis vulgaris. As a result, she was diagnosed as having sebopsoriasis. She was treated topically with an active vitamin D3 compound, 1a-24 (R) dihydroxycholecalciferol D3 (tacalcitol) cream. She applied tacalcitol cream twice daily for 4 weeks, and her facial eruptions thus cleared up completely. No recurrence was observed for 2 months thereafter, even though the use of tacalcitol cream was stopped. To investigate whether or not tacalcitol cream is generally effective for the treatment of such seborrheic dermatitis-like eruptions, three more patients were treated with tacalcitol cream. All patients exhibited scaly erythematous macules on the face and/or scalp, and their eruptions improved rapidly with tacalcitol cream. Tacalcitol cream was thus found to be effective and useful for the treatment of both sebopsoriasis and even seborrheic dermatitis of the face and scalp. PMID- 11056424 TI - Contact sensitization to corticosteroids: increased risk in long term dermatoses. AB - Patients affected by chronic dermatoses are at high risk for the development of sensitization to corticosteroids. This study was carried out to evaluate contact hypersensitivity to corticosteroids in a selected group of patients affected by longlasting cutaneous dermatoses. Sixty subjects underwent the GIRDCA series. The Italian GIRDCA series we applied had the following substances in addition to the European standard series: imidazolidinyl urea 2% pet, thiomersal 0.1% pet, disperse yellow 31% pet, disperse red 1% pet, 4'4-diaminodiphenylmethane 0.5% pet, ammoniated mercury 1% pet. The patients were tested with our corticosteroids series and some of them also with their own steroid products. Allergic reactions to corticosteroids were observed in 8/60 (13.3%) patients; budesonide was the main sensitizer (7 positives) followed by hydrocortisone-17-butyrate and betamethasone-17-valerate (2 and 1 positives, respectively). One patient also reacted to methylprednisolone aceponate contained in her own cream. 4/8 positives were certainly related to previous drug-exposure. This study showed a very high percentage of sensitization to corticosteroids. In our opinion, this value may be due both to our selection-standards and to the wide employment of a well known sensitizer like budesonide in our country. Although a corticosteroids series like ours seems to be adequate for the detection of sensitized patients, patch tests with individual compounds and/or steroids corresponding to local prescription habits are recommended, especially in unresponsive chronic dermatoses. PMID- 11056425 TI - Lichen striatus: clinical features and follow-up in 12 patients. AB - Lichen striatus (LS) is an uncommon disease of unknown origin characterised by a linear inflammatory papular eruption with spontaneous regression. We here review a series of 12 consecutive cases of LS. Diagnosis was supported by histological examination. Ten of our 12 patients were children aged 6 months to 12 years. The male gender predominated by 9:3. The lower limb was involved more often than the upper limb and trunk. The duration of the disease until regression ranged from 4 months to 4 years (median, 12 months). Postinflammatory hypopigmentation was noted in 5, and hyperpigmentation in 4 patients. Two patients showed nail involvement (onychodystrophy, longitudinal ridging) which appeared simultaneously with the skin lesions and resolved completely. A personal history of atopic disorders was found in 7 of 12 patients. From this series we can confirm that LS mainly affects children. Both skin and nail lesions disappear completely even if they last longer than one year. Compared with other studies, our series showed differences with regard to sex ratio, predilection sites and after effects. PMID- 11056426 TI - Lipoatrophic lesions preceded by pain and erythema - a new clinical entity? AB - A 46-year-old female patient presented with deeply depressed lesions on the abdominal wall after slight pain and erythema, without a history of trauma or injection. No general symptoms were noted. There was a marked decrease of subcutaneous fatty tissue with minimal inflammatory cell infiltration. We found three similar cases in the literature. Differential diagnoses were performed, but our case and the other three cases do not fit any of the heretofore described lipoatrophic diseases, and might be a new clinical entity. PMID- 11056427 TI - Spiny keratoderma of the palms and soles - report of two cases. AB - We report two patients with spiny keratoderma of the palms and soles characterized by multiple tiny keratotic plugs on the palms and soles. This disease was reported to be possibly associated with internal malignancies. We found a tumor from the esophagus to cardia in one patient. Another had no tumor but the lesion occurred soon after a severe bronchial asthma attack. Causal relation between spiny keratoderma of the palms and soles and bronchial asthma is obscure. Since this disease has been under-diagnosed and under-reported, it is important for dermatologists to keep spiny keratoderma of the palms and soles in mind in daily clinical examinations. PMID- 11056428 TI - Pemphigus vulgaris occurring simultaneously on a recent and an old surgical scar due to a Koebner's phenomenon. AB - Pemphigus vulgaris is an autoimmune bullous disorder involving the skin and sometimes the mucosa. Koebner's phenomenon is encountered when the typical features of a dermatosis are observed on a part of the skin previously subject to friction or trauma. A few cases of pemphigus vulgaris developing after damage to the skin and especially scars have been described. To the best of our knowledge, we report the first case where typical lesions of pemphigus vulgaris appeared simultaneously on a recent and an old scar as the sign of the reactivation of the disease. PMID- 11056429 TI - Stevens-Johnson syndrome-like exanthema secondary to methotrexate histologically simulating acute graft-versus-host disease. AB - A 61 year old male patient suffering from psoriasis vulgaris developed a severe skin reaction with toxic myelosuppression three days after administration of 20 mg methotrexate (MTX) p.o. per week and concomitant 100 mg acetylic salicylic acid (ASA) per day. The skin lesions simulated Stevens-Johnson syndrome with ulcerations of the oral mucosa and erythema multiforme-like target lesions. The histology of the epidermis resembled an acute graft-versus-host reaction. The increased toxic effect of MTX on keratinocytes in our patient was most likely caused by a lowered plasma binding capacity and reduced renal excretion of MTX due to concomitant administration of ASA. Thus in the treatment of severe forms of psoriasis with MTX, the combined administration of drugs aggravating MTX toxicity, particularly of ASA, should be carefully considered, due to the increased toxicity and risk of severe skin reactions. PMID- 11056430 TI - Cerebriform plantar hyperplasia: ultrastructural study of two cases. AB - In the present work we report the histopathological features of the cerebriform plantar hyperplasia observed in two patients with a mild form of the Proteus syndrome. Light microscopy revealed increased fibro-adipose tissue and adnexal structures in the dermis. Ultrastructurally, densely packed collagen fibrils variable in diameter and configuration, described as composite fibrils and unraveled fibrils, as well as a few fragmented elastic fibrils presenting an altered ratio between the elastin and the microfibrillar components were the major features observed. We consider that these histopathological findings will contribute to further delineate cerebriform plantar hyperplasia and also to establish clues for the early diagnosis of the Proteus syndrome. PMID- 11056431 TI - Angiosarcoma arising on rhinophyma. AB - We report an 82-year-old man who presented with a tumor which had developed over the previous year on the right nasal ala of a rhinophyma. Histopathological, immunohistochemical, and electron microscopic study confirmed the diagnosis of angiosarcoma on the head and neck. He was treated with radiotherapy of the tumor and cervical adenopathy, which developed later. The possible etiological and pathogenetic role of lymphedema due to inflammatory flares of rosacea on the nose is discussed, together with the histological and immunohistochemical data leading to the diagnosis of this tumor. PMID- 11056432 TI - Guess what! Human West African trypanosomiasis with chancre presentation. PMID- 11056433 TI - Guess what! Telangiectasia macularis eruptiva perstans involving the upper arms in an adult male. PMID- 11056434 TI - Guess what! Tinea manus during interferon-alpha therapy for chronic hepatitis C. PMID- 11056435 TI - Sexually transmitted diseases in the elderly. Review of 28 cases. AB - Sir, People over the age of 65 represent an increasingly large proportion of the population. In fact, ageing brings about some decrease in sexual arousal and activity [1] related to physical illness or medication needed to control a health condition [2]. Nevertheless, it is encouraging to find out that many elderly continue to be sexually active until their sixties, seventies and even eighties [3]. Information concerning the incidence of sexually transmitted infections in old people is sparse. Six retrospective studies have been published before [4-9]. They confirmed that the incidence of sexually transmitted diseases in patients over 65 years was significant for both newly acquired disease and for residual complications. Beyond that, there are sporadic case reports of HIV infection in older persons [10, 11], emphasising the persistence of risk behaviour in this population. PMID- 11056436 TI - Photodynamic therapy in dermatology. AB - Application of non-ionising radiation with or without photosensitizers is rather common in dermatology. Though the method itself was described in ancient times, its routine use in medicine based on scientific research started in the second half of the 20th century. Light can be used in three different patterns: phototherapy (UV-A or UV-B light), photochemotherapy (combination of psoralens with UV-A light) and photodynamic therapy (combination of photosensitizers with UV- and/or visible light). The following article deals with the photodynamic therapy or PDT. Using PDT implies the understanding of light dosimetry and calculation of light dose using different light sources and photosensitizers. The number of PDT sensitisers under investigation is rapidly increasing. The PDT itself, being a relatively new modality, quickly spreads its list of applications covering new indications in different areas of medicine. Though the main part of this list is made up of dermatological conditions, the use of PDT in other disciplines is also discussed to make dermatologists familiar with different aspects of the issue. PDT, like any treatment modality, has its benefits and adverse effects. The future of PDT is closely related to teamwork in physical, biochemical and clinical research which could provide better understanding of underlying mechanisms and help to create protocols for higher therapeutic efficacy. PMID- 11056437 TI - Hairless guinea pig skin: anatomical basis for studies of cutaneous biology AB - We would like to apologise to Dr. Sueki for the misplaced Figure 1 in his article Hairless guinea pig skin: anatomical basis for studies of cutaneous biology published in EJD vol. 10/5, page 359. Please find hereunder the correct Figure 1. PMID- 11056438 TI - Insulin resistance and elective surgery. PMID- 11056439 TI - Chylous ascites: a collective review. PMID- 11056440 TI - Clinicopathologic evaluation of hepatocellular carcinoma with bile duct thrombi. AB - BACKGROUND: The aim of this study was to evaluate the clinicopathologic characteristics of patients with hepatocellular carcinoma (HCC) and bile duct thrombi (BDT). PATIENTS: Seventeen patients with HCC and BDT among 671 patients with HCC who underwent hepatic resection were enrolled in this study. RESULTS: There were no significant differences in the survival rates between patients with and those without BDT, although the rate of stage IV or portal vein invasion was significantly higher in patients with HCC and BDT than in those with HCC but without BDT. In 9 of 17 patients with BDT, preoperative jaundice was observed. Five of the 17 patients underwent a bile duct resection combined with hepatic resection, and 12 patients underwent hepatic resection with removal of the BDT without bile duct resection. None of the patients had histopathologic evidence of direct tumor invasion into the bile duct wall or of any tumor recurrence related to the BDT. There were no significant differences in the survival rates between patients who underwent bile duct resection and those who did not. CONCLUSION: Hepatic resection and the removal of BDT without bile duct resection were sufficient surgical interventions to treat patients with HCC and BDT. PMID- 11056441 TI - Laparoscopic repair of chronic intrathoracic gastric volvulus. AB - BACKGROUND: Totally intrathoracic gastric volvulus is an uncommon presentation of hiatal hernia, in which the stomach undergoes organoaxial torsion predisposing the herniated stomach to strangulation and necrosis. This may occur as a surgical emergency, but some patients present with only chronic, non-specific symptoms and can be treated electively. The aim of this study is to describe a comprehensive approach to laparoscopic repair of chronic intrathoracic gastric volvulus and to critically assess the pre-operative work-up. METHODS: Eight patients (median age, 71 years) underwent complete laparoscopic repair of chronic intrathoracic gastric volvulus. Symptoms of epigastric pain and early satiety were universally present. Five patients had reflux symptoms. The diagnostic evaluation included a video esophagogram, upper endoscopy, 24-hour pH measurement, and esophageal manometry in all patients. Operative results and postoperative outcome were recorded and follow-up at 1 year included a barium swallow in all patients. RESULTS: All patients had documented intrathoracic stomach. Five of 8 patients had a structurally normal lower esophageal sphincter. All 4 patients with reflux esophagitis on upper endoscopy had a positive 24-hour pH study, and 2 of these patients had a structurally defective lower esophageal sphincter on manometry. None of the patients had preoperative evidence of esophageal shortening. All procedures were completed laparoscopically. The procedure included reduction of the stomach into the abdomen, primary closure of the diaphragmatic defect, and the construction of a short, floppy Nissen fundoplication. There were no major complications. One patient required repair of a trocar site hernia 6 months postoperatively. At 1-year follow-up, there were no radiologic recurrences of the volvulus. One patient complained of temporary swallowing discomfort and another had recurrent gastroesophageal reflux disease (GERD) symptoms caused by a breakdown of the wrap. All other patients remained asymptomatic during follow-up. CONCLUSIONS: The repair of chronic gastric volvulus can be accomplished successfully with a laparoscopic approach. A preoperative endoscopy and esophagogram are crucial to detect esophageal stricture or shortening, and manometry is needed to access esophageal motility; pH measurements do not affect operative strategy. The procedure should include a Nissen fundoplication to treat preoperative GERD, to prevent possible postoperative GERD, and to secure the stomach in the abdomen. The procedure is safe but technically challenging, requiring previous laparoscopic foregut surgical expertise. PMID- 11056442 TI - Clinical benefits of steroid therapy on surgical stress in patients with esophageal cancer. AB - BACKGROUND: Despite improvements in surgical techniques and perioperative care, severe complications lead to long hospital stays for some esophageal cancer patients. The purpose of this study was to evaluate the safety and effectiveness of perioperative steroid therapy on the postoperative clinical course. METHODS: Fifty-seven patients operated for esophageal cancer in 1997 and 1998 were treated with perioperative steroid therapy. Fifty consecutive patients operated in 1995 and 1996 served as a control group. In the steroid group, each patient was given 250 mg of methylprednisolone intravenously before operation followed by 125 mg on postoperative days 1 and 2. Serum interleukin-6, polymorphonuclear cell elastase, and C-reactive protein levels, and the postoperative clinical course were compared between the groups. RESULTS: Morbidity rates including hyperbilirubinemia, anastomotic leakage, and liver dysfunction were significantly lower in the steroid group than in the control group. Days until extubation and hospital stay were significantly shorter for the steroid group. Inflammatory mediators, body temperature, heart rate, and respiratory index after the surgical procedure were significantly lower in the steroid group. Adverse effects possibly caused by steroid therapy were not observed. CONCLUSIONS: Perioperative steroid therapy was safe and effective for the inhibition of inflammatory mediators and the improvement of the postoperative clinical course of patients with esophageal cancer. PMID- 11056443 TI - Effect of laparotomy and laparoscopy on the establishment of lung metastasis in a murine model. AB - BACKGROUND: Laparoscopic surgery is now applied to patients with gastrointestinal cancer. In animal studies, extraperitoneal tumor growth has been significantly less after laparoscopy than after laparotomy, but whether hematogenous metastasis occurs less frequently after laparoscopy is unknown. The aim of this study was to compare the frequency and growth of lung metastasis and serum levels of IL-6 and tumor necrosis factor-alpha (TNF-alpha) in mice treated by laparotomy and in mice treated by laparoscopy. METHODS: We used 182 male BALB/c mice. Colon 26 cancer cells (5 x 10(4)) were injected into the tail vein, and the mice were assigned to a laparotomy group (3-cm laparotomy), a laparoscopy group (carbon dioxide pneumoperitoneum at 6 to 8 mm Hg for 30 minutes), or a control group. Lung weight, number of lung metastases, and serum levels of IL-6 and TNF-alpha were measured and compared among the 3 groups. RESULTS: The lung weight and number of metastases on the lung surface and cut section in the laparotomy group (0.44+/ 0.21 g, 55.7+/-46.7, 23.0+/-19.0) were significantly larger than those in the laparoscopy group (0.32+/-0.15 g, 29.9+/- 25.5, 13.1+/-9.9) or the control group (0.28+/-0.13, 29.3+/-26.2, 11.1+/-11.1). Three hours after the procedures, the serum level of IL-6 was significantly higher in the laparotomy group (1353 +/- 790 pg/mL) than in the laparoscopy group (671+/-353 pg/mL) or the control group (333+/-341 pg/mL). The lung weight, number of lung metastases, and levels of IL-6 and TNF-alpha were not different between the laparoscopy and control groups. CONCLUSIONS: Our results indicate that, although laparotomy accelerates tumor metastasis to the lung in this murine model, laparoscopy does not increase the frequency and growth of lung metastasis. The laparoscopic approach may suppress hematogenous metastasis to the lung because of decreased surgical stress and reduced cytokine response. PMID- 11056444 TI - Macrophages infiltrating the tissue in chronic pancreatitis express the chemokine receptor CCR5. AB - BACKGROUND: The immunologic mechanisms involved in the development of chronic pancreatitis (CP) are poorly understood. Chronically inflamed tissues contain increased numbers of mononuclear cells expressing the CC chemokine receptor 5 (CCR5), which is also a coreceptor for HIV entry of macrophagetropic strains. However, whether this receptor is involved in the inflammatory process in CP is not known. In the current study, we analyzed the expression of CCR5 in CP. The detection of chemokine receptors on inflammatory cells would strongly suggest their involvement in the pathogenesis of CP (i.e., attraction and activation of these cells). To further evaluate this, we consecutively analyzed the expression of 2 ligands of CCR5: RANTES and MIP-alpha. METHODS: Pancreatic tissue samples of 22 patients with CP and of 7 healthy pancreas were evaluated. CCR5, RANTES, and MIP-1alpha were analyzed by Northern blot analysis. Consecutive tissue sections were stained for CCR5, CD3, and CD68 to define the leukocyte subtype expressing CCR5 in CP. RESULTS: By Northern blot analysis, CCR5, RANTES, and MIP-1alpha messenger RNA (mRNA) levels were 12.9-fold, 13.3-fold and 9.2-fold higher in CP specimens compared with healthy controls, respectively (P<.01). Immunostaining for CCR5 revealed a 30-fold increase of CCR5-positive cells in CP tissue compared with the healthy pancreas. Staining of consecutive tissue sections revealed that the majority of CCR5-positive cells were also CD68-positive (macrophages). CONCLUSIONS: Our data indicate that a remarkable portion of CCR5-positive cells in CP are macrophages. CCR5 is most likely involved in the attraction and activation of these macrophages, since the CCR5 ligands RANTES and MIP-1alpha are concomitantly upregulated. PMID- 11056445 TI - Hyperoxic reperfusion exacerbates postischemic renal dysfunction. AB - BACKGROUND: Hyperoxic reperfusion from global ischemia worsens functional outcome because of oxygen radical-mediated injury. This study tested the hypothesis that hyperoxic reperfusion would exacerbate postischemic renal dysfunction. METHODS: Twenty-nine healthy, uninephrectomized, male mongrel rabbits (Oryctolagus cuniculus) in 3 groups were subjected to 30 minutes of complete normothermic renal ischemia followed by reperfusion under hyperoxic or normoxic conditions. The groups were: hyperoxically reperfused (n = 8), normoxically reperfused (n = 8), hyperoxic sham (no ischemia, n = 5), and allopurinol-pretreated (50 mg/kg, intravenously), hyperoxically reperfused animals (n = 8). Plasma concentrations of creatinine, urea nitrogen and electrolytes were measured at 0, 24, 48, and 72 hours after ischemia and served as functional outcome markers. Histopathologic analysis of kidneys for injury was performed by an expert who was blinded to the procedures. RESULTS: Plasma creatinine in hyperoxically reperfused rabbits was significantly elevated above normoxic (P =.02) and sham (P =.003) animals by 48 hours and remained elevated to 72 hours. Plasma urea nitrogen in hyperoxically reperfused rabbits was significantly elevated above the normoxic group (P = .01), the sham group (P = .02), and the allopurinol group (P = .04) by 72 hours. These coincided with a significantly elevated histopathologic injury score in hyperoxically reperfused rabbits compared with sham (P = .019), normoxic (P = .035), and allopurinol-pretreated hyperoxically reperfused animals (P = .037). CONCLUSIONS: Hyperoxic reperfusion exacerbates renal dysfunction after 30 minutes of complete normothermic ischemia. This dysfunction may be mediated by oxygen radical-related injury. PMID- 11056446 TI - Anatomical segmentectomy of the head of the pancreas along the embryological fusion plane: a feasible procedure? AB - BACKGROUND: Less extensive resection of the head of the pancreas has been the procedure of choice recently for low-grade malignant neoplasms. The anatomical detail of the head of the pancreas is currently insufficient for segmental resection along the embryological fusion plane. METHODS: The anatomy of the head of the pancreas was analyzed in 31 consecutive autopsy specimens. An anterior (n = 10) or posterior (n = 10) segmentectomy of the head of each pancreas was performed along the macroscopically found fusion plane. The pancreatic arteries, the portal vein, the bile duct, and the pancreatic duct were visualized by injecting 3 silicon dyes of different colors. Another 11 specimens were examined by pancreatography before and after anterior (n = 5) or posterior (n = 6) segmentectomy. Eight of these 11 specimens were stained immunohistochemically to reveal the distribution of pancreatic polypeptide cells after segmentectomy. RESULTS: The cleavage between the anterior and posterior segments was discovered at the anterior inferior edge or at the posterior superior edge of the head of the pancreas. Anterior segmentectomy was accomplished while preserving the anterior and posterior pancreaticoduodenal arcades and the lower bile duct in the posterior segment. Posterior segmentectomy involved the removal of the lower bile duct and the posterior pancreaticoduodenal arcades. Pancreatography after segmentectomy showed the division of the ducts of Wirsung and Santorini with the peripheral branches. The immunohistochemical boundary of pancreatic polypeptide cells coincided with the surgical plane. These results showed the anterior and posterior segments were originated from the embryologically dorsal and ventral primordia, respectively. CONCLUSIONS: The current anterior or posterior segmentectomy of the head of the pancreas corresponded to the resection of the embryologically dorsal or ventral primordium, respectively. Anterior segmentectomy of the head of the pancreas might be a clinically applicable procedure; however, posterior segmentectomy involving the resection of the lower bile duct may be impractical. PMID- 11056447 TI - Pancreas divisum--really. PMID- 11056448 TI - Intra-abdominal cystic lymphangioma. PMID- 11056449 TI - Application of quality improvement to surgical practice. PMID- 11056450 TI - Invited commentary: application of quality improvement to surgical practice--a word of caution. PMID- 11056451 TI - Impact of breast cancer treatment guidelines on surgeon practice patterns: results of a hospital-based intervention. AB - BACKGROUND: Despite evidence regarding the effectiveness of post-surgical treatments for early-stage breast cancer, older women are less likely to receive appropriate therapy. We evaluated the impact of surgeon-specific "performance reports" on adherence to treatment guidelines among older women with breast cancer. METHODS: We obtained diagnostic and treatment data from hospital tumor registries supplemented with self-reported adjuvant therapy information on 1099 patients with stage I or II breast cancer diagnosed between November 1, 1992, and January 31, 1997, at 6 Rhode Island hospitals. We compared rates of appropriate treatment receipt before and after distribution of performance reports. Hierarchical analysis was used to account for the nesting of patients within surgeons. Separate analyses of mastectomy and breast-conserving surgery were performed. RESULTS: Age was negatively associated with post-surgical treatment, with patients who had breast-conserving surgery and who were older than 80 years significantly less likely to undergo radiation therapy (adjusted odds ratio = 0.08 [0.04, 0.14]) or appropriate adjuvant therapies (adjusted odds ratio = 0.14 [0.08, 0.22]) or both relative to 70- to 79-year-old patients. This effect did not improve post-intervention. While there was much variability in compliance with guidelines, surgeons' characteristics did not explain this variation. CONCLUSIONS: In Rhode Island, advanced age continues to be associated with less than adequate breast cancer therapy. Providing surgeons with "feedback" on the appropriateness of adjuvant treatment for older patients was insufficient to alter established practices. Using guideline compliance data as standard "quality indicators" of physician practice may be required. PMID- 11056452 TI - Invited commentary: surgical practice and decision making-is the patient's voice heard? PMID- 11056453 TI - Invited commentary: early-stage breast cancer treatment for elderly women-does one size fit all? PMID- 11056454 TI - See one, do one, teach one. PMID- 11056455 TI - On the uniform use of the AJCC/UICC staging system for hepatocellular carcinoma. PMID- 11056456 TI - Lichtenstein or plug-and-mesh in inguinal hernia repair? PMID- 11056457 TI - Reply PMID- 11056458 TI - Positive end expiratory pressure and response to inhaled nitric oxide therapy. PMID- 11056459 TI - Prolonged preservation increases complications after pancreas transplants. PMID- 11056460 TI - Comparative study on the distribution patterns of P2X(1)-P2X(6) receptor immunoreactivity in the brainstem of the rat and the common marmoset (Callithrix jacchus): association with catecholamine cell groups. AB - The present study investigated the topographical distribution of P2X(1)-P2X(6) receptor subtypes in the rat and common marmoset hindbrain by immunohistochemistry. In addition, double-labeling immunofluorescence was used to determine the extent of colocalization between catecholamine cell groups and the various P2X receptors. The data demonstrate a widespread distribution pattern for all six P2X receptors throughout both the rat hindbrain and the marmoset hindbrain, although distinctions between species, brain nuclei, and P2X receptor subtypes exist. In rat, dense staining for the P2X receptors was found in the nucleus of the solitary tract (NTS), medial vestibular nucleus, and medial and lateral parabrachial nuclei. Moderate staining was observed in the hypoglossal nucleus, cuneate nucleus, inferior olive, prepositus hypoglossi, rostral ventrolateral medulla (RVLM), and locus coeruleus. Staining was also observed in the gracile nucleus, the mesencephalic trigeminal nucleus, and the central pontine gray. In marmoset, prominent P2X receptor-like immunoreactivity occurred in the NTS, medial cuneate nucleus, prepositus hypoglossi, and medial vestibular nucleus. Moderate staining was observed in the area postrema, dorsal motor nucleus of the vagus, lateral cuneate, lateral reticular, spinal trigeminal nucleus, RVLM, and inferior olive. Immunofluorescent double labeling of tyrosine hydroxylase (TH)-containing cells revealed that all subtypes of P2X receptors show some degree of colocalization with TH. The highest proportion of TH and P2X receptor double labeling was in the A5 region (with the P2X(2) subunit), whereas the lowest proportion of double-labeled cells occurred in the C2 region of the NTS for the P2X(5) subunit. These findings support a role for extracellular adenosine 5'-triphosphate in fast synaptic neurotransmission within the brainstem. PMID- 11056461 TI - Nitric oxide synthase localized in a subpopulation of vestibular efferents with NADPH diaphorase histochemistry and nitric oxide synthase immunohistochemistry. AB - Efferent innervation of the vestibular labyrinth is known to be cholinergic. More recent studies have also demonstrated the presence of the neuropeptide calcitonin gene-related peptide in this system. Nitric oxide is one of a new class of neurotransmitters, the gaseous transmitters. It acts as a second messenger and neurotransmitter in diverse physiological systems. We decided to investigate the anatomical distribution of the synthetic enzyme for nitric oxide, nitric oxide synthase (NOS), to clarify the role of nitric oxide in the vestibular periphery. NADPH diaphorase histochemical and NOS I immunohistochemical studies were done in the adult chinchilla and rat vestibular brainstem; diaphorase histochemistry was done in the chinchilla periphery. Retrograde tracing studies to verify the presence of NOS in brainstem efferent neurons were performed in young chinchillas. Our light microscopic results show that NOS I, as defined mainly by the presence of NADPH diaphorase, is present in a subpopulation of both brainstem efferent neurons and peripheral vestibular efferent boutons. Our ultrastructural results confirm these findings in the periphery. NADPH diaphorase is also present in a subpopulation of type I hair cells, suggesting that nitric oxide might be produced in and act locally upon these cells and other elements in the sensory epithelium. A hypothesis about how nitric oxide is produced in the vestibular periphery and how it may interact with other elements in the vestibular sensory apparatus is presented in the discussion. PMID- 11056462 TI - Rostrocaudal nuclear relationships in the avian medulla oblongata: a fate map with quail chick chimeras. AB - We present a correlative fate map of the nonsegmented caudal hindbrain down to the medullospinal boundary (medulla oblongata), as a companion to a previous fate mapping study of the hindbrain rhombomeres r2-r6 in quail chick chimeras at stages HH10/11 [Marin and Puelles (1995) Eur J Neurosci 7:1714-1738]. For reproducibility and equivalent precision of analysis, successive portions of the medulla-called pseudorhombomeres "r7" to "r11"-were delimited by transverse planes through the center of adjacent somites at stages HH10/11. These units were each grafted homotopically and isochronically from quail donors into chick hosts. The chimeric specimens were fixed at stages HH35/36 and alternate Nissl-stained sagittal sections were compared to adjacent sections in which quail cells were detected immunocytochemically. This analysis in general showed that there is little intermixing between adjacent pseudorhombomeric domains, although some neuronal populations in the vestibular and trigeminal columns, as well as in the reticular formation and pontine nuclei, do migrate selectively into the host hindbrain. Contralateral migration was scarce up to the stages examined. Several motor nuclei, i.e., the vagal motor complex, or sensory nuclei, i.e., the medial vestibular nucleus, show cytoarchitectonic limits that coincide with pseudorhombomeric ones; however, most conventional grisea were found to originate across several pseudorhombomeres. The inferior olivary complex originated between "r8" and "r11" (between the centers of somites 1 and 5). The medullospinal boundary coincided precisely with the center of the fifth somite, slightly caudal to the obex and the end of the choroidal roof, and correlated with the end of many medullary cytoarchitectonic units. In contrast, the dorsal column nuclei and the caudal subnucleus of the descending trigeminal column fell within the spinal cord. On the whole, the patterns observed were very similar to those found before within the overtly segmented part of the hindbrain, suggesting that some underlying common mechanism may account for the transverse cytoarchitectonic boundaries. PMID- 11056463 TI - Molecular characterization and gene expression in the eye of the apolipophorin II/I precursor from Locusta migratoria. AB - The transport of lipids via the circulatory system of animals constitutes a vital function that uses highly specialized lipoprotein complexes. In insects, a single lipoprotein, lipophorin, serves as a reusable shuttle for the transport of lipids between tissues. We have found that the two nonexchangeable apolipoproteins of lipophorin arise from a common precursor protein, apolipophorin II/I (apoLp II/I). To examine the mechanisms of transport of lipids and liposoluble substances inside the central nervous system, this report provides the molecular cloning of a cDNA encoding the locust apoLp-II/I. We have recently shown that this precursor protein belongs to a superfamily of large lipid transfer proteins (Babin et al. [1999] J. Mol. Evol. 49:150-160). We determined that, in addition to its expression in the fat body, the locust apoLp-II/I is also expressed in the brain. Part of the signal resulted from fat body tissue associated with the brain; however, apoLp-II/I was strongly expressed and the corresponding protein detected, in pigmented glial cells of the lamina underlying the locust retina and in cells or cellular processes interspersed in the basement membrane. The latter finding strongly suggests an implication of apolipophorins in the transport of retinoids and/or fatty acids to the insect retina. PMID- 11056464 TI - Mouse cytomegalovirus in developing brain tissue: analysis of 11 species with GFP expressing recombinant virus. AB - Cytomegaloviruses (CMVs) are species-specific large double-stranded DNA viruses. Mouse and human CMVs have a similar morphology, similar gene sequence, and exert similar cellular effects, but the replication of the virus outside its primary host species is limited. This may confer upon CMV certain advantages for expression of foreign genes or cellular labels in brain cells of nonhost species. We examined the ability of recombinant mouse (m)CMV expressing green fluorescent protein (GFP) to serve as a vector for transgene expression in developing neurons and glia outside the normal host species. For comparative purposes, 11 species were examined. Mouse CMV reporter gene expression was particularly strong in the developing brain of its normal host species, mouse, where it replicated in cultures and brain slices, leading to cell death. All mammalian species tested (human, rat, gerbil, hamster, mouse) showed reporter gene expression after mCMV infection. High levels of mCMV infection were also found in chicken central nervous system cells in vitro, and a low level of mCMV expression was found after an initial delay in turtle neurons and glia. No mCMV reporter gene expression was found in frog cells or aplysia neurons or glia or in drosophila or fungal cells. Infection of nonmouse neurons by low concentrations of mCMV led to strong expression of GFP in dendrites and axons with normal morphology. Despite the lack of replication, high doses of mCMV induced morphologic changes in neurons and glia from hamster and rat brain slices, leading to cells rounding up, and to the formation of giant cells consisting of an aggregate of many cells fused together into a syncytium. In contrast, in human hippocampal slices, GFP-expressing cells infected with mCMV had a relatively normal appearance 12 days after inoculation. To determine whether a CMV from another species could serve as a vector for gene transfer, a recombinant human CMV-expressing GFP was used for transgene expression in rat brain cells in vitro. Cytomegaloviruses thus have potential as useful vectors for gene transfer and labeling central nervous system cells, with the actions of CMV being dependent on a number of factors. PMID- 11056465 TI - Postnatal development of nitric oxide synthase expression in the mouse superior colliculus. AB - Since nitric oxide has a role in the refinement of the retinal projection to the superior colliculus (SC), we studied the onset of neuronal nitric oxide synthase (nNOS) expression in the mouse SC in order to compare its development with that of the refinement process. Sections from animals at ages P1, P5, P8, P11, P15, and P21 and adults were examined with nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) histochemistry or immunocytochemistry using an antibody directed against nNOS. At all ages there was a wedge of labeled neurons in the dorsolateral periaqueductal gray extending into the deep layers of the SC. At P1 there was also a single superficial band of labeled neurons within the region that will become the intermediate gray layer (IGL). By P5, labeled neurons were also seen in what will become the superficial gray layer. There was a ventral to dorsal progression in nNOS expression with substantial changes in the numbers of labeled neurons in the different laminae between P5 and adulthood. The number of labeled neurons in the IGL peaked at P15, whereas in the superficial layers the numbers continued to increase through P21 and then declined in adults. At all ages these neurons represented a variety of morphological cell types. The onset of nNOS expression in the different laminae is earlier than has been reported in studies using NADPHd as a marker for nNOS. The temporal and spatial patterns of nNOS expression reported here match more closely the time course of pathway refinement in the SC, providing additional evidence for the involvement of nitric oxide in this process. PMID- 11056466 TI - Topography and associations of luteinizing hormone-releasing hormone and neuropeptide Y-immunoreactive neuronal systems in the human diencephalon. AB - Neuropeptide Y (NPY) potentiates the effect of luteinizing hormone-releasing hormone (LHRH) on luteinizing hormone secretion in several species, including human. In addition to the pituitary sites, the interactions of the NPY and LHRH systems may involve diencephalic loci. However, the morphologic basis of this putative communication has not yet been elucidated in the human brain. To discover interaction sites, the distribution and connections of LHRH and NPY immunoreactive (IR) neuronal elements in the human hypothalamus were investigated by means of light microscopic single- and double-label immunocytochemistry. NPY IR perikarya and fibers were found to be widely distributed in the ventral diencephalon, with high densities in the preopticoseptal, periventricular, and tuberal regions. Small neuronal cell groups were infiltrated with a dense network of varicose NPY-IR fibers in the lateral preoptic area. The LHRH-IR perikarya were located mainly in the preopticoseptal region, diagonal band of Broca, lamina terminalis, and periventricular and infundibular nuclei. A few LHRH-IR neurons and fibers were scattered in the mamillary region. The overlap between the NPY and LHRH systems was apparent in the periventricular, paraventricular, and infundibular nuclei. Double-labeling immunohistochemistry showed NPY-IR axon varicosities in contact with LHRH-IR perikarya and main dendrites. The putative innervation of LHRH neurons by NPY-IR fibers was also seen in 1-microm-thick plastic sections and with confocal laser scanning microscope, thus further supporting the functional impact of NPY-IR terminals on LHRH-IR neurons. The present findings suggest that the hypophysiotropic LHRH-synthesizing neurons may be innervated by intrahypothalamic NPY-IR fibers. Confirmation by ultrastructural analysis would demonstrate that the LHRH system in the human hypothalamus is regulated by NPY, as has been demonstrated in nonhuman species. PMID- 11056467 TI - Choline acetyltransferase-immunoreactive neurons in the developing rat retina. AB - The development of cholinergic cells in the rat retina has been examined with immunocytochemistry by using antisera against choline acetyltransferase (ChAT). ChAT-immunoreactive (IR) cells were first detected at embryonic day 17 (E17) in the transitional zone between the neuroblastic layer (NBL) and ganglion cell layer (GCL). At E20, ChAT-IR cells are located exclusively in the GCL. At postnatal day 0 (P0), ChAT immunoreactivity appeared for the first time in cells at the distal margin of the NBL. Two prominent bands of labeled processes were first visible at P3, and by P15, these two bands resembled those of the adult retina. In addition, ChAT immunoreactivity appeared transiently in horizontal cells from P5 to P10. The number of ChAT-IR cells increased steadily up to P15. This resulted in a 93.8-fold increase between E17 and P15 (680-63,800 cells). However, after P15, the number declined by 19% from 63,800 cells at P15 to 51,800 in the adult. At all ages, the spatial density of each ChAT-IR cell population in the central retina was higher than in the periphery. In both central and peripheral regions, the peak density of ChAT-IR cells in the GCL was attained at E20. However, in the INL, the peak densities occurred at P3 in the central region and at P5 in the peripheral region. Up to P15, the soma diameter of ChAT-IR cells in the INL and GCL in each region increased continuously, reaching peak values at P15. Our results demonstrate that ChAT immunoreactivity is expressed in early developmental stages in the rat retina, as in other mammals, and that acetylcholine released from ChAT-IR cells may have neurotrophic functions in retinal maturation. PMID- 11056468 TI - Effects of acute and chronic gonadectomy on the catecholamine innervation of the cerebral cortex in adult male rats: insensitivity of axons immunoreactive for dopamine-beta-hydroxylase to gonadal steroids, and differential sensitivity of axons immunoreactive for tyrosine hydroxylase to ovarian and testicular hormones. AB - Previous studies have shown that gonadectomy in adult male rats induces a complex series of region- and time-specific changes in the density of presumed cerebral cortical dopamine axons that are immunoreactive for tyrosine hydroxylase. The present study asked whether noradrenergic cortical afferents also show hormone sensitivity by assaying axons immunoreactive for the enzyme dopamine-beta hydroxylase in representative areas of acutely and chronically gonadectomized and sham-operated adult male rats. Catecholamine afferents (both tyrosine hydroxylase immunoreactive and dopamine-beta-hydroxylase-immunoreactive) were also quantified in gonadectomized rats supplemented with testosterone propionate, with 17-beta estradiol, or with 5-alpha-dihydrotestosterone. Analyses of noradrenergic (dopamine-beta-hydroxylase) afferents revealed no differences in axon appearance or density among the hormonally intact and hormonally manipulated groups. However, analyses of tyrosine hydroxylase immunoreactivity revealed an unexpected division of labor among ovarian and testicular hormones in ameliorating the effects of acute verses chronic hormone deprivation on these afferents. Estradiol replacement attenuated the decreases in immunoreactivity induced by acute gonadectomy, but was ineffective in suppressing changes in immunoreactivity stimulated by chronic gonadectomy. In contrast, supplementing gonadectomized animals with dihydrotestosterone provided no protection from acute decreases in innervation, but fully attenuated both the supragranular decreases and infragranular increases in tyrosine hydroxylase-immunoreactive axon density that mark the association cortices of chronically gonadectomized rats. Together these findings indicate both long- and short-term effects of gonadectomy on cortical catecholamines, principally target dopamine afferents, and that chronic gonadectomy, which selectively disturbs dopamine innervation in the prefrontal cortices, involves a compromise in androgen signaling pathways. PMID- 11056469 TI - Reduced synaptic clustering of GABA and glycine receptors in the retina of the gephyrin null mutant mouse. AB - Clustering of neurotransmitter receptors in postsynaptic densities involves proteins that aggregate the receptors and link them to the cytoskeleton. In the case of glycine and GABA(A) receptors, gephyrin has been shown to serve this function. However, it is unknown whether gephyrin is involved in the clustering of all glycine and GABA(A) receptors or whether it interacts only with specific isoforms. This was studied in the retinae of mice, whose gephyrin gene was disrupted, with immunocytochemistry and antibodies that recognize specific subunits of glycine and GABA(A) receptors. Because homozygous (geph -/-) mutants die around birth, an organotypic culture system of the mouse retina was established to study the clustering of gephyrin and the receptors in vitro. We found that all gephyrin and all glycine receptor clusters (hot spots) were abolished in the geph (-/-) mouse retina. In the case of GABA(A) receptors, there was a significant reduction of clusters incorporating the gamma2, alpha2, and alpha3 subunits; however, a substantial number of hot spots was still present in geph (-/-) mutant retinae. This shows that gephyrin interacts with all glycine receptor isoforms but with only certain forms of GABA(A) receptors. In heterozygous geph (+/-) mutants, no reduction of hot spots was observed in the retina in vivo, but a significant reduction was found in the organotypic cultures. This suggests that mechanisms may exist in vivo that allow for the compensation of a partial gephyrin deficit. PMID- 11056470 TI - Periaqueductal gray neurons monosynaptically innervate extranuclear noradrenergic dendrites in the rat pericoerulear region. AB - Previous reports using light microscopy have provided anatomical evidence that neurons in the ventrolateral periaqueductal gray (PAG) innervate the medial pericoerulear dendrites of noradrenergic neurons in the nucleus locus coeruleus (LC). The present study used anterograde tracing and electron microscopic analysis to provide more definitive evidence that neurons in the ventrolateral PAG form synapses with the somata or dendrites of noradrenergic LC neurons. Deposits of either biotinylated dextran amine or Phaseolus vulgaris leucoagglutinin into the rat ventrolateral PAG labeled a moderate to high number of axons in the region of the medial pericoerulear region and Barrington's nucleus, but a relatively low number were labeled in the nuclear core of the LC. Ultrastructural analysis of anterogradely labeled terminals at the levels of the rostral (n = 233) and caudal (n = 272) subdivisions of the LC indicated that approximately 20% of these form synapses with tyrosine hydroxylase-immunoreactive dendrites; most of these were located in the medial pericoerulear region. In rostral sections, about 12% of these were symmetric synapses, 9% were asymmetric synapses, and 79% were membrane appositions without clear synaptic specializations. In caudal sections, about 30% were symmetric synapses, 11% were asymmetric synapses, and 59% were appositions. In both rostral and caudal sections, 60% of the anterogradely labeled terminals formed synapses with noncatecholamine dendrites, and 20% formed axoaxonic synapses. These results provide direct evidence for monosynaptic projections from neurons in the ventrolateral PAG to the extranuclear dendrites of noradrenergic LC neurons. This monosynaptic pathway may mediate in part the analgesia, reduced responsiveness to external stimuli, and decreased excitability of somatic motoneurons produced by stimulation of neurons in the ventrolateral PAG. PMID- 11056471 TI - Catenins, Wnt signaling and cancer. AB - Recent studies indicate that plakoglobin may have a similar function to that of beta-catenin within the Wnt signaling pathway. beta-catenin is known to be an oncogene in many forms of human cancer, following acquisition of stabilizing mutations in amino terminal sequences. Kolligs(1) and coworkers show, however, that unlike beta-catenin, plakoglobin induces neoplastic transformation of rat epithelial cells in the absence of such stabilizing mutations. Cellular transformation by plakoglobin also appears to be distinct from that of beta catenin in that it requires activation of the proto-oncogene c-myc. Surprisingly, c-myc is activated more efficiently by plakoglobin than beta-catenin, despite its previous identification as a target of Tcf/beta-catenin.(2) In contrast, a synthetic Tcf reporter gene is activated to a much greater extent by beta-catenin than plakoglobin. Plakoglobin and beta-catenin may therefore have different roles in Wnt signaling and cancer, which reflect their differential effects on target gene activity. PMID- 11056472 TI - Closing the gaps among a web of DNA repair disorders. AB - As recently as six years ago, three human diseases with similar phenotypes were mistakenly believed to be caused by a single genetic defect. The three diseases, Ataxia-telangiectasia, Nijmegen breakage syndrome, and an AT-like disorder are now known, however, to have defects in three separate genes: ATM, NBS1, and MRE11. Furthermore, new recent studies have shown now that all three gene products interact; the ATM kinase phosphorylates NBS1, which, in turn, associates with MRE11 to regulate DNA repair. Remarkably or expectedly, depending on one's point of view, the similarity in disease phenotypes is evidently due to defects in a common DNA repair pathway. PMID- 11056473 TI - A neuro (endo)crine regulation of bone remodeling. AB - Bone remodeling is the normal physiologic process that is used by vertebrates to maintain a constant bone mass during the period bracketed by the end of puberty and the onset of gonadal failure in later life. Besides the well-characterized and critical process of local regulation of bone remodeling, achieved by autocrine and paracrine mechanisms, recent genetic studies have shown that there is a central control of bone formation, mediated by a neuroendocrine mechanism. This central regulation involves leptin, an adipocyte-secreted hormone that controls body weight, reproduction and bone remodeling, and which binds to and exerts its effect through the cells of the hypothalamic nuclei in the brain. This genetic result in mice is in line with clinical observations in humans and generates a whole new direction of research in bone physiology. BioEssays 22:970 975, 2000. PMID- 11056474 TI - Vertebrate anteroposterior patterning: the Xenopus neurectoderm as a paradigm. AB - This review discusses formation of the vertebrate anteroposterior (AP) axis, focusing on the dorsal ectoderm, which gives rise to the nervous system, using the frog Xenopus as a model. After summarizing classical models of AP neural patterning, we describe recent molecular studies that are encouraging re examination of these models. Such studies have shown that AP ectodermal patterning occurs by the onset of gastrulation, much earlier than previously thought. The identity of tissues that determine AP pattern is discussed, and the definition of the Organizer is reconsidered. The activity of factors secreted by inducing tissues in early patterning decisions is assessed and formulated into a revised model for Xenopus AP neural patterning. Finally, AP ectodermal patterning in Xenopus dorsal ectoderm is compared to that of other germ layers, and to other vertebrates. PMID- 11056475 TI - Neuregulin, a factor with many functions in the life of a schwann cell. AB - The signalling system comprising the ligand Neuregulin-1, and its receptors, ErbB2 and ErbB3, plays multiple and important roles in glial development. These include functions in early development of neural crest cells, in expansion of the Schwann cell precursor pool and in myelination. Neuregulin is one of the crucial axon-derived signals that influence development of Schwann cells. These are specialized cells that ensheath peripheral axons and provide electrical insulation. Schwann cells have also long been implicated in providing more than a simple ensheathing function. Compelling evidence for this has emerged from the analysis of mice lacking these cells, resulting from a non-functional or compromised Neuregulin signalling system. They serve as a model to study glia nerve interactions in vivo and indicate that Schwann cells provide important neurotrophic signals, and also cues that regulate perineurium development and nerve fasciculation. PMID- 11056476 TI - The puzzle of PCNA's many partners. AB - The identification of proteins that interact with proliferating cell nuclear antigen (PCNA) has recently been a rapidly expanding field of discovery. PCNA is involved in many aspects of DNA replication and processing, forming a sliding platform that can mediate the interaction of proteins with DNA. It is striking that many proteins bind to PCNA through a small region containing a conserved motif; these include proteins involved in cell cycle regulation as well as those involved in DNA processing. Sequential and regulated binding of motif-containing proteins to PCNA may contribute to the ordering of events during DNA replication and repair. Results from bacteriophages and archaea show that the structural basis for the interaction of this motif with PCNA is extremely ancient. The analysis of how such functional motifs have been recruited to proteins in present day organisms helps us to understand how these complex systems arose from ancestral organisms. PMID- 11056477 TI - Apoptosis in cancer: cause and cure. AB - The accumulation of neoplastic cells can occur through enhanced proliferation, diminished cell turnover, or a combination of both processes. Although the potential contribution of diminished cell turnover to tumor development has been appreciated for a decade, more recent studies in animal models and clinical cancer specimens have elucidated the mechanisms by which alterations in the apoptotic machinery contribute to the process of carcinogenesis. At the same time, a different group of studies have demonstrated the feasibility of eliminating neoplastic cells by selectively inducing apoptosis. In this essay, we review recent developments in the fields of carcinogenesis and molecular therapeutics in light of new understanding of apoptotic pathways. PMID- 11056478 TI - Random walks and cell size. AB - For many years, it has been believed that diffusion is the principle motive force for distributing molecules within the cell. Yet, our current information about the cell makes this improbable. Furthermore, the argument that limitations responsible for the relative constancy of cell size--which seldom varies by more than a factor of 2, whereas organisms can vary in mass by up to 10(24)--are based on the limits of diffusion is questionable. This essay seeks to develop an alternative explanation based on transport of molecules along structural elements in the cytoplasm and nucleus. This mechanism can better account for cell size constancy, in light of modern biological knowledge of the complex microstructure of the cell, than simple diffusion. PMID- 11056479 TI - Quality control in databanks for molecular biology. AB - Using a scientific measurement without an estimate of its error is like lending money to a stranger. Given the explosion in nucleic acid and protein sequence and structural data, what risks are the scientific and medical communities running in using these databases. Is there an 'ombudsman' who speaks for the users of the data? CODATA, the Committee on Data for Science and Technology of the International Council of Scientific Unions was established to improve the quality, reliability, processing, management, and accessibility of data for science and technology. The CODATA Task Group on Biological Macromolecules has surveyed quality control procedures of archival databanks in molecular biology. Our role is 'to advise, to be consulted, and to warn.' This report describes the kinds and extents of errors that may appear in nucleic acid and protein databases, and presents an agenda for future work to improve the quality of these databases. The results of the survey appear on the webhttp://www.codata.org/codata/tgreports/ tg_reps.html. PMID- 11056480 TI - The contractile vacuole and its membrane dynamics. AB - The contractile vacuole (CV) is an osmoregulatory organelle whose mechanisms of function are poorly understood. Immunological studies in the last decade have demonstrated abundant proton-translocating V-type ATPases (V-ATPases) in its membrane that could provide the energy, from proton electrochemical gradients, for moving ions into the CV to be followed by water. This review emphasizes recent work on the contractile vacuole complex (CVC) of Paramecium including (1) CV expulsion, (2) a role for V-ATPases in sequestering fluid, (3) identifying ions in the cytosol and in the CV, (4) in situ electrophysiological parameters of the CVC membrane, and (5) a better understanding of the membrane dynamics of this organelle. PMID- 11056481 TI - Modularity in development and evolution. PMID- 11056482 TI - From development to molecular medicine. PMID- 11056483 TI - Patterning a chemotactic response. PMID- 11056484 TI - Olson EN, williams RS. Remodeling muscles with calcineurin. Bioessays 2000;22:510 519 AB - The following sentence was omitted from the legend to Figure 3 in the above mentioned article: The schematic depiction of intracellular calcium waveforms in skeletal muscle myofibers was based on primary data acquired by E.R. Chin and D.G.Allen ((62)). The authors and the BioEssays editorial office would like to express their regrets to Drs. Chin and Allen for this omission. PMID- 11056485 TI - Endocrine treatment of prostate cancer PMID- 11056486 TI - The male climacterium: clinical signs and symptoms of a changing endocrine environment. AB - Frailty is characterized by generalized weakness, impaired mobility and balance, and poor endurance. Loss of muscle strength is an important factor in the process of frailty, and is the limiting factor for an individual's chances of living an independent life until death. In men, several hormonal systems show a decline in activity during aging. Serum bioavailable testosterone (T) and estradiol (E2), dehydroepiandrosterone (DHEA) and its sulfate (DHEAS), and growth hormone (GH) and insulin-like growth factor (IGF)-I concentrations all decrease during aging in men. Physical changes during aging have been considered physiologic, but there is evidence that some of these changes are related to this decline in hormonal activity. In a cross-sectional study performed among 403 independently living elderly men, positive independent associations were observed between serum bioavailable T and muscle strength and bone mineral density (BMD). Serum T was negatively associated with fat mass. Serum luteinizing hormone (LH) increased with age and was inversely associated with T. Independent of T, LH was negatively related with muscle strength and positively with the number of problems in activities of daily living. Further, a positive relation was present between serum E2 and BMD. A positive association between DHEAS and BMD was dependent on T and E2 concentrations. Finally, in the same study, men with the highest E2 concentrations were significantly more satisfied with life, measured with a questionnaire developed by Herschbach and Huber, compared to men with the lowest E2 concentrations. In conclusion, these findings are in agreement with other studies, which suggest that the maintenance of a good physical functional ability and quality of life is related to serum T, E2, and DHEA(S) concentrations. PMID- 11056487 TI - Age, libido, and male sexual function. AB - In the last decade of the 20th century, there was a distinct reappraisal of male sexual dysfunction and its pharmaco-medical treatment. Although representative studies of sexual (dys)function in the aging male (i.e., between 60-90 years of age) are still lacking, one might assume with certainty that many men and their partners could benefit from sexological counseling and treatment. At the same time, it is obvious that many older men with erectile dysfunction do not seek or want treatment for various reasons. Nevertheless, it is obligatory that modern physicians include sexual matters in dealing with their aging patients, as an essential part of their quality of life. The doctor of today should approach the old(er) male patient with sexual dysfunction (regardless of comorbidity) in an identical manner as young(er) patients, i.e., with proper sexological history taking, proper physical examination, and/or sexological diagnostic screening, and discussing the various available treatments. Needless to say, that they should not "create" sexual problems when patients are satisfied with their current way of life. However, with the increasing number of efficacious treatments, doctors will satisfy many of their older patients with sexual difficulties who seek treatment. PMID- 11056488 TI - The prostate as an endocrine organ: androgens and estrogens. PMID- 11056489 TI - Immediate vs. delayed androgen deprivation for prostate cancer. AB - Androgen ablation has been the standard treatment of symptomatic patients with metastatic prostate cancer for more than 50 years. Within the last 15 years, the introduction of prostate-specific antigen (PSA) has induced a stage migration toward less extensive disease and a dramatic decrease in the proportion of men presenting with N+/M+ disease. Historical studies, conducted during the pre-PSA era, are therefore of limited interest in counseling modern patients. The routine use of radical therapies such as radical prostatectomy and radiotherapy has considerably expanded the problem of timing of endocrine treatment in range and complexity. Advanced disease is now diagnosed in patients with limited involvement of extraprostatic sites and even in patients presenting an isolated elevation of PSA after radical treatment. In the absence of clear guidelines, data from past literature and ongoing modern studies were compiled in the present review in an attempt to generate practical considerations. PMID- 11056490 TI - Endocrine treatment: expected duration stage by stage. PMID- 11056491 TI - Side effects of endocrine treatment and their mechanisms: castration, antiandrogens, and estrogens. AB - Endocrine treatment of prostate cancer can be performed under several different regimes. They all have side effects which in different ways influence quality of life and the patient's general health. This paper is a survey of the most important early side effects of the different modes of endocrine treatment, their etiology, and possible ways to avoid or treat them. PMID- 11056492 TI - Potential side-effects of endocrine treatment of long duration in prostate cancer. AB - Endocrine treatment of prostate cancer has been established for more than 5 decades. Focusing on immediate or short-term side effects, bilateral orchidectomy may cause psychological trauma, treatment with oral estrogens is combined with a high risk of severe cardiovascular complications, and the use of LH-RH agonists and antiandrogens as monotherapies or in combination may result in tumor flare, hot flashes, and gynecomastia. In recent years an increasing number of reports on anemia and/or osteoporosis related to endocrine treatment have been published. These side effects are regular and persistent after orchidectomy, or during treatment with LH-RH agonists, and are most often expressed with maximum androgen blockade. In contrast, anemia and/or osteoporosis are not reported with estrogen treatment or the use of nonsteroidal antiandrogens as a monotherapy regimen. Since many prostate cancer patients are treated hormonally for many years, control of Hb levels and bone mineral density before and after initiation of treatment at regular intervals is highly recommended as a standard of care. PMID- 11056493 TI - Exploitable mechanisms for the blockade of androgenic action. PMID- 11056494 TI - Foreword PMID- 11056495 TI - Does induction of apoptosis contribute to the overall clinical profile of terazosin? Introduction. PMID- 11056496 TI - Pathology of benign prostatic hyperplasia. PMID- 11056497 TI - Development of a new in vitro model for the study of benign prostatic hyperplasia. AB - BACKGROUND: The search for novel agents for the treatment of the lower urinary tract symptoms (LUTS) associated with benign prostatic hyperplasia (BPH) is dependent on an increased understanding of the pathophysiology of the disease. Unfortunately, in vitro and animal models have been of limited value. METHODS: This article describes a novel model system in which the interactions of the stromal and epithelial components of the prostate gland can be determined. RESULTS: The coculture system provides a simple model of the cellular interactions occurring in the adult human prostate. CONCLUSIONS: This coculture system could potentially be used to determine the precise molecular site of action of agents such as terazosin on prostatic apoptosis. PMID- 11056498 TI - In vitro models of prostate apoptosis: clusterin as an antiapoptotic mediator. PMID- 11056499 TI - Regulation of apoptosis in prostatic disease. AB - BACKGROUND: Recent studies suggest that aberrant regulation of apoptosis, including acquired apoptosis resistance, contributes to perturbations in cell growth that, in part, underlie the development of benign prostatic hyperplasia (BPH) and prostate cancer. Importantly, apoptosis resistance can enhance the malignant properties of prostate cancer cells, contributing to their widespread metastatic activities. Since apoptosis resistance likely contributes to benign and malignant prostate disease, the promotion of apoptosis represents a reasonable therapeutic objective. METHODS: This brief review focuses on the role of apoptosis in prostatic disease and discusses potential intervention points for novel therapeutics. CONCLUSIONS: Novel therapies for prostatic disease, including gene therapy and biological therapy that involve apoptosis as a mechanism of action, are being developed and tested. Ultimately, the identification of proapoptotic and antiapoptotic genes and the pathways through which they operate will serve to provide more rational approaches for further development of more efficacious therapeutic strategies for benign and malignant prostatic disease. PMID- 11056500 TI - Alpha(1)-adrenoceptors (alpha(1)-AR) and vascular smooth muscle cell growth. AB - BACKGROUND: Smooth muscle cells in vascular tissue, like tissue within the urogenital sinus, undergo growth and proliferation. METHODS: This review attempts to compare and contrast the mechanisms and controlling factors involved in prostatic and vascular tissue. There is a particular focus on the role of catecholamines and alpha-adrenoceptors (alpha-ARs), and on the effects of alpha(1)-AR antagonists (blockers) on cellular dynamics. RESULTS AND CONCLUSIONS The situation in vascular tissue appears analagous to that in prostatic tissue. Certain AR-antagonists, in addition to altering smooth muscle contraction, may have other actions on cellular dynamics. PMID- 11056501 TI - Effects of alpha1-adrenoceptor antagonists on cultured prostatic smooth muscle cells. AB - BACKGROUND: alpha1-adrenoceptor (alpha1-AR) antagonists, used to relieve the lower tract urinary symptoms (LUTS) in benign prostate hyperplasia (BPH) patients, are thought to act in inhibiting the contraction of stromal smooth muscle. An attempt was made using new technology to visualize and quantify the effect of alpha1-AR antagonists in a cell culture model of prostatic smooth muscle cells (SMC). METHODS: Prostatic smooth muscle cells cultured from human prostate tissue were treated with alpha1-AR agonists and antagonists. The effects on cell growth, cell contraction, differentiation status, and apoptosis were determined by means of an MTT cell viability assay, time-lapse video microscopy, RT-PCR analysis, and FACS analysis of annexin V/propidium iodide-stained cells, respectively. RESULTS: Prostatic smooth muscle cells derived from prostate tissue expressed SMC-specific markers. They showed spontaneous contractions, and phenylephrine increased the percentage of contracting cells by 3-fold. alpha1-AR antagonists inhibited spontaneous as well as phenylephrine-induced contractions. Long-term treatment with doxazosin induced differentiation tended towards a contractile phenotype, as indicated by an increase of the ratio of smooth muscle heavy chain myosin subtypes SM2/SM1. There was, however, no effect on cell growth. High concentrations of antagonist (100 microM) induced apoptosis in about 80% of the treated SMC. This effect was not cell-type-specific and was also seen in skin fibroblasts and immortalized prostate epithelial cells. CONCLUSION: In an easy-to-handle cell culture model of prostatic smooth muscle cells, the effects of alpha1-AR antagonists on cell contraction, growth, and differentiation can be investigated. The results indicate that in addition to inhibition of cell contraction, alpha1-AR antagonists have the potential to induce apoptosis. PMID- 11056502 TI - Effects of alpha(1)-adrenoceptor (alpha(1)-AR) antagonists on cell proliferation and apoptosis in the prostate: therapeutic implications in prostatic disease. AB - BACKGROUND: Benign prostate hyperplasia (BPH) and prostate cancer established that disruption of the molecular mechanisms that regulate apoptosis and cell proliferation among the stromal and epithelial cell populations, may underlie the neoplastic development that characterizes the aging gland. This work examined the effects of selected alpha(1)-adrenoceptor (alpha(1)-AR) antagonists (blockers) on cellular dynamics to determine whether induction of apoptosis or inhibition of proliferation could contribute to the overall clinical profile. METHODS: Our efforts were focused on investigating whether alpha(1)-AR antagonists of two different chemical classes affect prostate pathophysiology via mechanisms other than smooth muscle contraction. In in vitro experiments, the two clinically used quinazoline alpha(1)-adrenoceptor antagonists terazosin and doxazosin and the chemically-distinct sulphonamide, tamsulosin, were examined for effects on prostatic tumor growth, by inhibiting cell proliferation and'or inducing apoptosis. RESULTS: Our findings suggest that alpha(1)-AR antagonists, terazosin and doxazosin, suppress prostatic growth by inducing apoptosis in a dose dependent manner and without affecting cell proliferation. Tamsulosin exerted no effect on prostate cancer cell growth. The apoptotic effect of terazosin and doxazosin appears to be independent of the alpha(1)-adrenoceptor block. CONCLUSIONS: Taken together, our findings demonstrate the ability of the quinazoline alpha-blockers, terazosin and doxazosin, but not the sulphonamide, tamsulosin, to suppress prostate growth by inducing apoptosis among the epithelial cells in the benign and malignant prostate. These studies underwrite the durability of the response seen in long-term studies with terazosin, and suggest the potential of this drug in the treatment of prostate carcinoma. PMID- 11056503 TI - Clinical perspective on apoptosis in the management of the BPH patient. AB - BACKGROUND: Traditionally, alpha(1)-adrenoceptor (AR) antagonists have been assumed to produce clinical benefit by an exclusive action on the tone of the periurethral stromal smooth muscle. However, recent evidence has emerged of additional intra- and extraprostatic actions. METHODS: This article attempts to put into clinical context the recently described effects of certain alpha(1)-AR on prostate cell dynamics (i.e., proliferation and apoptosis). RESULTS AND CONCLUSIONS There is good evidence that certain alpha(1)-AR antagonists, in addition to affecting stromal smooth muscle, have effects on prostatic apoptosis that contribute to the overall clinical profile. Furthermore, this is not a class effect and may be restricted to balanced quinazoline alpha blockers (BQABs), such as terazosin. PMID- 11056519 TI - In vivo function of class I myosins. PMID- 11056520 TI - PYK2 is an adhesion kinase in macrophages, localized in podosomes and activated by beta(2)-integrin ligation. AB - Pyk2 is a member of the focal adhesion kinase (FAK) family, highly expressed in the central nervous system and haemopoietic cells. Although Pyk2 is homologous to FAK, its role in signaling pathways was shown to be distinct from that of FAK. We show here that Pyk2 is highly expressed in peritoneal IC-21 macrophage and is tyrosine phosphorylated in response to cell attachment to fibronectin and fibrinogen. Upon IC-21 cell adhesion, Pyk2 tyrosine phosphorylation is inhibited by blocking antibodies to the integrin subunits alpha(M) and beta(2). Furthermore, Pyk2 is rapidly tyrosine phosphorylated in response to ligation of beta(2) integrins by antibodies. In migrating macrophages, Pyk2 localizes to perinuclear regions and to podosomes, where it is clustered with tyrosine phosphorylated proteins. Furthermore, in the podosomal ring structure, which surrounds the central actin core, Pyk2 co-localizes with vinculin, talin, and paxillin. In the podosomes, Pyk2 also co-localizes with the integrin alpha(M)beta(2). Lastly, reduction of Pyk2 expression in macrophages leads to inhibition of cell migration. We propose that Pyk2 is functionally linked to the formation of podosomes where it mediates the integrin-cytoskeleton interface and regulates cell spreading and migration. PMID- 11056521 TI - Sorting of tropomyosin isoforms in synchronised NIH 3T3 fibroblasts: evidence for distinct microfilament populations. AB - The nonmuscle actin cytoskeleton consists of multiple networks of actin microfilaments. Many of these filament systems are bound by the actin-binding protein tropomyosin (Tm). We investigated whether Tm isoforms could be cell cycle regulated during G0 and G1 phases of the cell cycle in synchronised NIH 3T3 fibroblasts. Using Tm isoform-specific antibodies, we investigated protein expression levels of specific Tms in G0 and G1 phases and whether co-expressed isoforms could be sorted into different compartments. Protein levels of Tms 1, 2, 5a, 6, from the alpha Tm(fast) and beta-Tm genes increased approximately 2-fold during mid-late G1. Tm 3 levels did not change appreciably during G1 progression. In contrast, Tm 5NM gene isoform levels (Tm 5NM-1-11) increased 2-fold at 5 h into G1 and this increase was maintained for the following 3 h. However, Tm 5NM-1 and -2 levels decreased by a factor of three during this time. Comparison of the staining of the antibodies CG3 (detects all Tm 5NM gene products), WS5/9d (detects only two Tms from the Tm 5NM gene, Tm 5NM-1 and -2) and alpha(f)9d (detects specific Tms from the alpha Tm(fast) and beta-Tm genes) antibodies revealed 3 spatially distinct microfilament systems. Tm isoforms detected by alpha(f)9d were dramatically sorted from isoforms from the Tm 5NM gene detected by CG3. Tm 5NM-1 and Tm 5NM-2 were not incorporated into stress fibres, unlike other Tm 5NM isoforms, and marked a discrete, punctate, and highly polarised compartment in NIH 3T3 fibroblasts. All microfilament systems, excluding that detected by the WS5/9d antibody, were observed to coalign into parallel stress fibres at 8 h into G1. However, Tms detected by the CG3 and alpha(f)9d antibodies were incorporated into filaments at different times indicating distinct temporal control mechanisms. Microfilaments in NIH 3T3 cells containing Tm 5NM isoforms were more resistant to cytochalasin D-mediated actin depolymerisation than filaments containing isoforms from the alpha Tm(fast) and beta-Tm genes. This suggests that Tm 5NM isoforms may be in different microfilaments to alpha Tm(fast) and beta-Tm isoforms even when present in the same stress fibre. Staining of primary mouse fibroblasts showed identical Tm sorting patterns to those seen in cultured NIH 3T3 cells. Furthermore, we demonstrate that sorting of Tms is not restricted to cultured cells and can be observed in human columnar epithelial cells in vivo. We conclude that the expression and localisation of Tm isoforms are differentially regulated in G0 and G1 phase of the cell cycle. Tms mark multiple microfilament compartments with restricted tropomyosin composition. The creation of distinct microfilament compartments by differential sorting of Tm isoforms is observable in primary fibroblasts, cultured 3T3 cells and epithelial cells in vivo. PMID- 11056522 TI - Nitric oxide modulates intracellular translocation of pigment organelles in Xenopus laevis melanophores. AB - Pigment organelles in Xenopus laevis melanophores are used by the animal to change skin color, and they provide a good model for studying intracellular organelle transport. Movement of organelles and vesicles along the cytoskeleton is essential for many processes, such as axonal transport, endocytosis, and intercompartmental trafficking. Nitric oxide (NO) is a signaling molecule that plays a role in, among other things, relaxation of blood vessels, sperm motility, and polymerization of actin. Our study focused on the effect NO exerts on cytoskeleton-mediated transport, which has previously received little attention. We found that an inhibitor of NO synthesis, N-nitro-L-arginine methyl ester (L NAME), reduced the melatonin-induced aggregation of the pigment organelles, melanosomes. Preaggregated melanosomes dispersed after treatment with L-NAME but not after exposure to the inactive stereoisomer (D-NAME) or the substrate for NO synthesis (L-arginine). Signal transduction by NO can be mediated through the activation of soluble guanylate cyclase (sGC), which leads to increased production of cGMP and activation of cGMP-dependent kinases (PKG). We found that both the sGC inhibitor 1H-(1,2,4) oxadiazolo(4,3-a)quinoxalin-1-one (ODQ) and the cGMP analogue 8-bromoguanosine 3':5'-cyclic monophosphate (8-Br-cGMP) reduced melanosome aggregation, whereas the PKG inhibitor KT582 did not. Our results demonstrate that melanosome aggregation depends on synthesis of NO, and NO deprivation causes dispersion. It seems, thus, as if NO and cGMP are essential and can regulate melanosome translocation. PMID- 11056523 TI - Distribution of polyglutamylated tubulin in the flagellar apparatus of green flagellates. AB - Polyglutamylation is a widely distributed posttranslational modification of tubulin that can be demonstrated either by biochemical analysis or by the use of specific antibodies like GT335. Western blotting using GT335 demonstrated that polyglutamylated tubulin is enriched in isolated basal apparatus of Spermatozopsis similis. Single- and double-labeling experiments, using indirect immunofluorescence and immunogold electron microscopy of isolated cytoskeletons of S. similis and Chlamydomonas reinhardtii, revealed that polyglutamylated tubulin was predominately present in the basal bodies and the proximal part of the axonemes. Using immunogold labeling of whole mounts of Spermatozopsis cytoskeletons, we obtained evidence for a predominant occurrence of polyglutamylated tubulin in the B-tubule of the axonemal doublets. Polyglutamylation occurs early during premitotic basal body assembly in S. similis, whereas the probasal bodies of Chlamydomonas, which are present through interphase, showed a reduced staining with GT335 indicating that polyglutamylation is involved in basal body maturation. During flagella regeneration of C. reinhardtii, polyglutamylation preceded detyrosination and became visible shortly after the onset of flagellar regeneration. In C. reinhardtii and S. similis polyglutamylated tubulin was absent or highly reduced in the flagellar transition region, a specialized part of the flagellum linking the basal body to the axoneme. Furthermore, the transition region and the neighboring part of the axoneme showed reduced staining with L3, an antibody directed against detyrosinated tubulin. The results indicate that differences in the modification pattern can occur in a confined area of individual microtubules. The deficiency of polyglutamylated and detyrosinated tubulin in the transition region could have functional implications for flagellar turnover or excision. PMID- 11056524 TI - Induction of Alzheimer-specific Tau epitope AT100 in apoptotic human fetal astrocytes. AB - In Alzheimer's and other neurodegenerative diseases, hyperphosphorylated tau accumulates in affected neuronal and glial cells in the form of paired helical filaments (PHFs). This tau binds antibody AT100, which recognizes the double phosphorylation site (Thr212/Ser214) that is not present in normal biopsy tau. In primary cultures, highly enriched (>98%) in astrocytes of human fetal brain, three polypeptides of 52, 64, and 70 kD showed immunoreactivity with tau antibodies against non-phosphorylated epitopes, accounting for 88, 12, and <1%, respectively, of the total reactivity. All three polypeptides were phosphorylated at the PHF-1 epitope but not at the epitopes Tau-1, 12E8, AT8, and AT100. Treatment of cultures with okadaic acid resulted in apoptosis characterized by the blebbing of the plasma membrane, condensation of nuclear chromatin, and fragmentation of the nucleus. This treatment also resulted in a 3- to 5-fold increase in the content of both tau protein and phosphorylation. The increases were observed in all phosphorylation sites examined, and included the AT100 site. The AT100 site has been proposed to be generated by protein kinase B/Akt and Cdc2. Since okadaic acid can induce an AD-like hyperphosphorylated state of normal tau in primary cultures of human brain cells, a simple cellular model is available permitting study of self-aggregation of tau and phosphorylation events characteristic of neurodegeneration. PMID- 11056530 TI - Regulation of cytoplasmic dynein behaviour and microtubule organization by mammalian Lis1. AB - Whereas total loss of Lis1 is lethal, disruption of one allele of the Lis1 gene results in brain abnormalities, indicating that developing neurons are particularly sensitive to a reduction in Lis1 dosage. Here we show that Lis1 is enriched in neurons relative to levels in other cell types, and that Lis1 interacts with the microtubule motor cytoplasmic dynein. Production of more Lis1 in non-neuronal cells increases retrograde movement of cytoplasmic dynein and leads to peripheral accumulation of microtubules. These changes may reflect neuron-like dynein behaviours induced by abundant Lis1. Lis1 deficiency produces the opposite phenotype. Our results indicate that abundance of Lis1 in neurons may stimulate specific dynein functions that function in neuronal migration and axon growth. PMID- 11056531 TI - Drosophila Lis1 is required for neuroblast proliferation, dendritic elaboration and axonal transport. AB - Haplo-insufficiency of human Lis1 causes lissencephaly. Reduced Lis1 activity in both humans and mice results in a neuronal migration defect. Here we show that Drosophila Lis1 is highly expressed in the nervous system. Lis1 is essential for neuroblast proliferation and axonal transport, as shown by a mosaic analysis using a Lis1 null mutation. Moreover, it is cell-autonomously required for dendritic growth, branching and maturation. Analogous mosaic analysis shows that neurons containing a mutated cytoplasmic-dynein heavy chain (Dhc64C) exhibit phenotypes similar to Lis1 mutants. These results implicate Lis1 as a regulator of the microtubule cytoskeleton and show that it is important for diverse physiological functions in the nervous system. PMID- 11056532 TI - A role for the lissencephaly gene LIS1 in mitosis and cytoplasmic dynein function. AB - Mutations in the LIS1 gene cause gross histological disorganization of the developing human brain, resulting in a brain surface that is almost smooth. Here we show that LIS1 protein co-immunoprecipitates with cytoplasmic dynein and dynactin, and localizes to the cell cortex and to mitotic kinetochores, which are known sites for binding of cytoplasmic dynein. Overexpression of LIS1 in cultured mammalian cells interferes with mitotic progression and leads to spindle misorientation. Injection of anti-LIS1 antibody interferes with attachment of chromosomes to the metaphase plate, and leads to chromosome loss. We conclude that LIS1 participates in a subset of dynein functions, and may regulate the division of neuronal progenitor cells in the developing brain. PMID- 11056533 TI - Molecular model of a lattice of signalling proteins involved in bacterial chemotaxis. AB - Coliform bacteria detect chemical attractants by means of a membrane-associated cluster of receptors and signalling molecules. We have used recently determined molecular structures, in conjunction with plastic models generated by three dimensional printer technology, to predict how the proteins of the complex are arranged in relation to the plasma membrane. The proposed structure is a regular two-dimensional lattice in which the cytoplasmic ends of chemotactic-receptor dimers are inserted into a hexagonal array of CheA and CheW molecules. This structure creates separate compartments for adaptation and downstream signalling, and indicates a possible basis for the spread of activity within the cluster. PMID- 11056534 TI - Cadherin-mediated regulation of microtubule dynamics. AB - Epithelial polarization and neuronal outgrowth require the assembly of microtubule arrays that are not associated with centrosomes. As these processes generally involve contact interactions mediated by cadherins, we investigated the potential role of cadherin signalling in the stabilization of non-centrosomal microtubules. Here we show that expression of cadherins in centrosome-free cytoplasts increases levels of microtubule polymer and changes the behaviour of microtubules from treadmilling to dynamic instability. This effect is not a result of cadherin expression per se but depends on the formation of cell-cell contacts. The effect of cell-cell contacts is mimicked by application of beads coated with stimulatory anti-cadherin antibody and is suppressed by overexpression of the cytoplasmic cadherin tail. We therefore propose that cadherins initiate a signalling pathway that alters microtubule organization by stabilizing microtubule ends. PMID- 11056535 TI - cAMP-GEFII is a direct target of cAMP in regulated exocytosis. AB - Although cAMP is well known to regulate exocytosis in many secretory cells, its direct target in the exocytotic machinery is not known. Here we show that cAMP GEFII, a cAMP sensor, binds to Rim (Rab3-interacting molecule, Rab3 being a small G protein) and to a new isoform, Rim2, both of which are putative regulators of fusion of vesicles to the plasma membrane. We also show that cAMP-GEFII, through its interaction with Rim2, mediates cAMP-induced, Ca2+-dependent secretion that is not blocked by an inhibitor of cAMP-dependent protein kinase (PKA). Accordingly, cAMP-GEFII is a direct target of cAMP in regulated exocytosis and is responsible for cAMP-dependent, PKA-independent exocytosis. PMID- 11056536 TI - Functional dissection of in vivo interchromosome association in Saccharomyces cerevisiae. AB - Homologue pairing mediates both recombination and segregation of chromosomes at meiosis I. The recognition of nucleic-acid-sequence homology within the somatic nucleus has an impact on DNA repair and epigenetic control of gene expression. Here we investigate interchromosomal interactions using a non-invasive technique that allows tagging and visualization of DNA sequences in vegetative and meiotic live yeast cells. In non-meiotic cells, chromosomes are ordered in the nucleus, but preferential pairing between homologues is not observed. Association of tagged chromosomal domains occurs irrespective of their genomic location, with some preference for similar chromosomal positions. Here we describe a new phenomenon that promotes associations between sequence-identical ectopic tags with a tandem-repeat structure. These associations, termed interchromosome trans associations, may underlie epigenetic phenomena. PMID- 11056537 TI - The apoptotic v-cyclin-CDK6 complex phosphorylates and inactivates Bcl-2. AB - v-cyclin encoded by Kaposi's sarcoma herpesvirus/human herpesvirus 8 (KSHV or HHV8) associates with cellular cyclin-dependent kinase 6 (CDK6) to form a kinase complex that promotes cell-cycle progression, but can also induce apoptosis in cells with high levels of CDK6. Here we show that whereas HHV8-encoded v-Bcl-2 protects against this apoptosis, cellular Bcl-2 has lost its anti-apoptotic potential as a result of an inactivating phosphorylation in its unstructured loop region. Moreover, we identify Bcl-2 as a new substrate for v-cyclin-CDK6 in vitro, and show that it is present in a complex with CDK6 in cell lysates. A Bcl 2 mutant with a S70A S87A double substitution in the loop region is not phosphorylated and provides resistance to apoptosis, indicating that inactivation of Bcl-2 by v-cyclin-CDK6 may be required for the observed apoptosis. Furthermore, the identification of phosphorylated Bcl-2 in HHV8-positive Kaposi's sarcoma indicates that HHV8-mediated interference with host apoptotic signalling pathways may encourage the development of Kaposi's sarcoma. PMID- 11056538 TI - Function of microtubules in intercellular transport of plant virus RNA. AB - Cell-to-cell progression of tobacco mosaic virus (TMV) infection in plants depends on virus-encoded movement protein (MP). Here we show that a conserved sequence motif in tobamovirus MPs shares similarity with a region in tubulins that is proposed to mediate lateral contacts between microtubule protofilaments. Point mutations in this motif confer temperature sensitivity to microtubule association and viral-RNA intercellular-transport functions of the protein, indicating that MP-interacting microtubules are functionally involved in the transport of vRNA to plasmodesmata. Moreover, we show that MP interacts with microtubule-nucleation sites. Together, our results indicate that MP may mimic tubulin assembly surfaces to propel vRNA transport by a dynamic process that is driven by microtubule polymerization. PMID- 11056539 TI - PAN, the proteasome-activating nucleotidase from archaebacteria, is a protein unfolding molecular chaperone. AB - The proteasome-activating nucleotidase (PAN) from Methanococcus jannaschii is a complex of relative molecular mass 650,000 that is homologous to the ATPases in the eukaryotic 26S proteasome. When mixed with 20S archaeal proteasomes and ATP, PAN stimulates protein degradation. Here we show that PAN reduces aggregation of denatured proteins and enhances their refolding. These processes do not require ATP hydrolysis, although ATP binding enhances the ability of PAN to prevent aggregation. PAN also catalyses the unfolding of the green fluorescent protein with an 11-residue ssrA extension at its carboxy terminus (GFP11). This unfolding requires ATP hydrolysis, and is linked to GFP11 degradation when 20S proteasomes are also present. This unfolding activity seems to be essential for ATP-dependent proteolysis, although PAN may function by itself as a molecular chaperone. PMID- 11056540 TI - The effect of Golgi depletion on exocytic transport. AB - Here we evaluate the idea that the Golgi is in dynamic equilibrium with the endoplasmic reticulum (ER). In cytoplasts that lack the Golgi apparatus, no regrowth of the Golgi is observed, nor is any transport from the ER to the cell surface detected. However, introduction of the smallest measurable amount of Golgi (equivalent to a few per cent per cell) yields significant exocytic transport. Our results indicate that the steady-state levels of Golgi in the ER are far smaller than the 30% that has been postulated, and that the Golgi may be an independent organelle and not simply an extension of the ER. PMID- 11056541 TI - Glycine 384 is required for presenilin-1 function and is conserved in bacterial polytopic aspartyl proteases. AB - Endoproteolysis of beta-amyloid precursor protein (betaAPP) and Notch requires conserved aspartate residues in presenilins 1 and 2 (PS1 and PS2). Although PS1 and PS2 have therefore been proposed to be aspartyl proteases, no homology to other aspartyl proteases has been found. Here we identify homology between the presenilin active site and polytopic aspartyl proteases of bacterial origin, thus supporting the hypothesis that presenilins are novel aspartyl proteases. PMID- 11056542 TI - Mutations in the Plk gene lead to instability of Plk protein in human tumour cell lines. AB - It has been established that mutations in Drosophila Polo cause abnormalities in mitosis. In human cells, maximal Plk activity is reached in the M phase of the cell cycle, and the function of Plk is therefore considered to be required for mitotic cellular events such as spindle formation, chromosome segregation and cytokinesis. Microinjection of anti-Plk antibody into living cells has been found to induce a mitotic abnormality that contributes to the generation of aneuploidy, and this is an important finding in relation to tumour development. Indeed, previous studies have shown that the level of expression of a mitotic checkpoint gene, hsMAD2, is reduced and that another checkpoint gene, BUB1, is mutated in certain human cancer cells. PMID- 11056543 TI - Type II myosin regulatory light chain relieves auto-inhibition of myosin-heavy chain function. AB - The F-actin based motor protein myosin II has a key role in cytokinesis. Here we show that the Schizosaccharomyces pombe regulatory light chain (RLC) protein Rlc1p binds to Myo2p in manner that is dependent on the IQ sequence motif (the RLC-binding site), and that Rlc1p is a component of the actomyosin ring. Rlc1p is important for cytokinesis at all growth temperatures and is essential for this process at lower temperatures. Interestingly, all deleterious phenotypes associated with the loss of Rlc1p function are suppressed by deletion of the RLC binding site on Myo2p. We conclude that the sole essential function of RLCs in fission yeast is to relieve the auto-inhibition of myosin II function, which is mediated by the RLC-binding site, on the myosin heavy chain (MHC). PMID- 11056544 TI - Primary cells suppress oncogene-dependent apoptosis. AB - Oncogenes that promote cell-cycle progression also sensitize cells to agents that induce apoptosis, possibly by inactivating inhibitors that ordinarily provide protection against cell death. Here we show that the adenoviral oncogene E1A sensitizes cells to an anti-cancer drug by at least two pathways. One establishes a link between the drug and pro-apoptotic factors, but is not sufficient for sensitization without the second pathway, which suppresses inhibitors of apoptosis. PMID- 11056545 TI - ELSO - not just another meeting. European Life Scientist Organization. PMID- 11056546 TI - Model is as model does. PMID- 11056547 TI - An Arrow straight to the heart of Wingless signalling. PMID- 11056548 TI - A rough guide to a smooth brain. PMID- 11056549 TI - Cyclin' on the viral path to destruction. PMID- 11056550 TI - Plant microtubules meet their MAPs and mimics. PMID- 11056551 TI - Signalling for stability. PMID- 11056552 TI - Erratum. PMID- 11056553 TI - Conformational disease. AB - A large and diverse number of diseases are now recognized as 'conformational diseases', caused by adoption of non-native protein conformations that lead to aggregation. The recent conference, 'Alpha1-antitrypsin deficiency and other conformational diseases', held in Airlie, Virginia, USA (27-30 June, 2000) focused on some of the common pathways by which cells protect themselves from toxicity associated with protein misfolding and aggregation. PMID- 11056555 TI - The professional debunker. PMID- 11056554 TI - Fly methods for the new millennium. PMID- 11056556 TI - Human herpesvirus 8 virology, epidemiology and related diseases. AB - Human herpesvirus 8 ([HHV-8], Kaposi's sarcoma-associated herpesvirus [KSHV]) is a novel human oncovirus classified as a gamma-herpesvirus. HHV-8 is associated with Kaposi's sarcoma (KS), primary effusion lymphoma (PEL) and some cases of multicentric Castleman's disease (MCD). Antibodies against HHV-8 are detected in the sera of almost all KS patients, about 30% of HIV-infected homosexual males in the world and 1.4% of the Japanese population. In HHV-8-associated malignancies such as KS and PEL, HHV-8 latently infects these tumor cells. Unlike other viruses, HHV-8 encodes several human homologues including cytokines (IL-6, MIPs, IRFs) and regulatory proteins (cyclin D, G-protein coupled receptor [GPCR]). These proteins may play significant roles in the pathogenesis of HHV-8-associated diseases. It has been demonstrated in vitro that the functions of retinoblastoma and p53 proteins were inhibited by viral cyclin D and latency-associated nuclear antigen, respectively. Mice transgenic for GPCR have a KS-like region in the skin. These data suggest the full oncogenecity of HHV-8. On the other hand, many cells expressing lytic proteins are found in MCD tissues, suggesting that the pathogenesis of MCD is different from that of HHV-8-associated malignancies. PMID- 11056557 TI - Jackfruit lectin: properties of mitogenicity and the inhibition of herpesvirus infection. AB - Jackfruit lectin (JFL) from Artocarpus heterophyllus has been found to exhibit inhibitory activity in vitro with a cytopathic effect towards herpes simplex virus type 2 (HSV-2), varicella-zoster virus (VZV), and cytomegalovirus (CMV). The 50% inhibitory dose values from plaque reduction assay (inactivation) were 2.5, 5, and 10 Eg/ml of JFL for HSV-2, VZV, and CMV, respectively. Lymphocyte proliferation was significantly increased in the presence of the JFL in the concentration range of 2.5 to 50 Eg/ml, but was reduced at 500 Eg/ml. It was found that CD16(+)/CD56(+) cells (natural killer cells) were induced among the primary lymphocyte subpopulations. The activity of natural killer (NK) cells was not affected by JFL in the concentration range of 5 to 500 Eg/ml. These data suggest that JFL is mitogenic for NK lymphocyte (CD16(+)/CD56(+)) and also active against HSV-2, VZV, and CMV. PMID- 11056558 TI - Prevalence of spotted fever rickettsial antibodies in dogs and rodents in the Philippines. AB - Antibodies against spotted fever group rickettsiae have been detected in blood samples of dogs and rodents obtained from selected areas in the Philippines. In this serosurvey, the positive percentage rates are 8.3% (11/132) in dogs and 12.2% (6/49) in rats. Positive results were read from samples tested with Rickettsia japonica antigen. No positive result was obtained in blood samples of rats and house mice using R. akari antigen. The findings of this study are the first to confirm the detection of spotted fever group rickettsial antibodies in the Philippines. PMID- 11056559 TI - An increase in multi-drug-resistant isolates of Salmonella typhimurium from healty carriers in Aichi, Japan. AB - To investigate the prevalence of drug-resistant isolates of Salmonella Typhimurium in Aichi, Japan, we performed antimicrobial susceptibility tests for 148 isolates from healthy carriers, and from sporadic and outbreak cases of salmonellosis from 1980 to 1999. We found an increase in drug-resistant isolates from 56% (37/66) in the 1980s to 74% (61/82) in the 1990s due to increasing examples of four-, five-, and six-drug resistances. Of 98 resistant isolates in 1980-1999, 12 were identified as ampicillin (A)-, chloramphenicol (C)-, streptomycin (S)-, sulfonamide (Su)-, and tetracycline (T)-resistant S. Typhimurium (4 in the 1980s, 8 in the 1990s), whose pattern was identical to that of multi-drug-resistant S. Typhimurium definitive phage type 104 (DT104) which has been recently detected in various developed countries. Six-drug-resistance ACSSuTP (piperacillin), in which P was added to the core pattern of the ACSSuT, was also found in four isolates in the 1980s and seven in the 1990s. Another six drug-resistant pattern, ACSSuTN (nalidixic acid), appeared in five isolates in the 1990s. These multi-drug-resistant isolates were predominately found in healthy carriers (21/28), suggesting that in Aichi the multi- (five- or six-) drug-resistant isolates of S. Typhimurium have existed in healthy carriers as well as in diarrhea patients in 1980 to 1999. PMID- 11056560 TI - Highly active antiretroviral therapy for the treatment of Kaposi's sarcoma associated with primary human immunodeficiency virus type-1 infection. PMID- 11056561 TI - "Cephem-induced vancomycin resistance" observed in phenotypically methicillin sensitive Staphylococcus aureus isolated from a patient treated with vancomycin and cephems. PMID- 11056562 TI - Influenza C viruses isolated during the 1999-2000 influenza season in Saitama Prefecture, Japan. PMID- 11056563 TI - Successful treatment of acute myeloradiculoneuritis with high-dose corticosteroids in a patient with primary HIV-1 infection. PMID- 11056564 TI - Isolation of influenza C virus during the 1999/2000-influenza season in Hiroshima Prefecture, Japan. PMID- 11056565 TI - Outbreaks of heat stable enterotoxin-producing Escherichia coli O169 in the Kinki district in Japan: genotypic comparison by pulsed-field gel electrophoresis of isolates from two outbreaks in 2000 with isolates from four outbreaks in 1997 1998. PMID- 11056566 TI - Has echinococcus granulosus settled in Hokkaido? PMID- 11056567 TI - Isolation and molecular comparison of Japanese encephalitis virus in Ishikawa, Japan. PMID- 11056568 TI - A piece of my mind: financial indigestion. PMID- 11056569 TI - JAMA 100 years ago: RECEPTIVE QUIESCENCE OF THE STOMACH DURING MASTICATION PMID- 11056570 TI - Evolution of the fire ant lesion. PMID- 11056571 TI - Sleep disorders, often unrecognized, complicate many physical illnesses. PMID- 11056572 TI - Medical errors vs medical injuries: physicians seek to prevent both. PMID- 11056574 TI - From the Food and Drug Administration. PMID- 11056575 TI - From the food and drug administration: new drug for HIV infection PMID- 11056576 TI - From the food and drug administration: new treatment for leukemia PMID- 11056577 TI - Barriers to patients seeking emergency care for acute coronary heart disease. PMID- 11056578 TI - Barriers to patients seeking emergency care for acute coronary heart disease PMID- 11056579 TI - Deficiencies in US medical care. PMID- 11056580 TI - Deficiencies in US medical care. PMID- 11056581 TI - Deficiencies in US medical care. PMID- 11056582 TI - Deficiencies in US medical care. PMID- 11056583 TI - How many deaths are due to medical errors? PMID- 11056584 TI - How many deaths are due to medical errors? PMID- 11056585 TI - How many deaths are due to medical errors? PMID- 11056586 TI - How many deaths are due to medical errors? PMID- 11056587 TI - Lead in candle wicks. PMID- 11056588 TI - Lead in candle wicks PMID- 11056589 TI - Chemical analysis of ecstasy pills. PMID- 11056590 TI - Evaluation of HIV-1 immunogen, an immunologic modifier, administered to patients infected with HIV having 300 to 549 x 10(6)/L CD4 cell counts: A randomized controlled trial. AB - CONTEXT: Despite enormous improvements achieved through the use of antiretroviral therapies (ARTs), the risk for eventual human immunodeficiency virus (HIV) disease progression remains high. Agents that enhance the immunologic mechanism for viral recognition might reduce disease progression. OBJECTIVE: To determine whether the addition of HIV-1 Immunogen would confer added clinical efficacy to that achievable by ARTs. DESIGN AND SETTING: Multicenter, double-blind, placebo controlled, randomized trial beginning March 1996 and ending May 1999 conducted at 77 centers in the United States providing primary care or referral care for persons infected with HIV. PATIENTS: Adults infected with HIV who have baseline CD4 cell counts between 300 x 10(6)/L and 549 x 10(6)/L without prior acquired immunodeficiency syndrome-defining conditions receiving stable ART (or no therapy) were screened and 2527 were randomized. INTERVENTIONS: Ten units of HIV 1 Immunogen, derived from a Zairian HIV isolate, inactivated and formulated with incomplete Freund adjuvant, was administered intramuscularly every 12 weeks. The placebo was incomplete Freund adjuvant. Changes in ARTs were allowed. MAIN OUTCOME MEASURES: HIV progression-free survival; secondary end points included overall survival, changes in HIV RNA, CD4 cell counts, CD4 percentage, body weight, and immunogenicity. RESULTS: The overall event rate was 1.8 per 100 person-years of follow-up. Fifty-three subjects developed clinical progression in each treatment group (relative risk [RR], 0.97; 95% confidence interval [CI], 0.66-1.42; P =.89). There were 19 and 23 deaths in the placebo and HIV-1 Immunogen groups, respectively (RR, 0.81; 95% CI, 0.44-1. 48; P =.49). There were no statistically significant differences between the groups with respect to changes in HIV RNA (P =.59), CD4 percentage (P =.63), or body weight (P =.89). Subjects in the HIV-1 Immunogen group had an increase in average CD4 cell count of approximately 10 x 10(6)/L greater than the placebo group (P =.02). CONCLUSION: HIV-1 Immunogen with unrestricted ART failed to demonstrate an increase in HIV progression-free survival. JAMA. 2000;284:2193-2202. PMID- 11056591 TI - Policies on faculty conflicts of interest at US universities. AB - CONTEXT: Despite federal regulations on faculty conflicts of interest in federally funded research, academic-industry ties are common, and evidence exists that financial considerations bias the research record. Public scrutiny of these ties is increasing, especially in cases where researchers have financial interests in the corporate sponsors of their clinical research. OBJECTIVE: To review policies on conflict of interest at major biomedical research institutions in the United States. DESIGN: Cross-sectional survey and content analysis study conducted from August 1998 to February 2000. SETTING AND PARTICIPANTS: The 100 US institutions with the most funding from the National Institutes of Health in 1998 were initially sampled; policies from 89 institutions were available and included in the analysis. MAIN OUTCOME MEASURES: Process for disclosure, review, and management of conflicts of interest and specified management strategies or limitations, according to the institutions' faculty/staff conflict of interest policies. RESULTS: Content of the conflict of interest policies varied widely across institutions. Fifty-five percent of policies (n = 49) required disclosures from all faculty while 45% (n = 40) required them only from principal investigators or those conducting research. Nineteen percent of policies (n = 17) specified limits on faculty financial interests in corporate sponsors of research, 12% (n = 11) specified limits on permissible delays in publication, and 4% (n = 4) prohibited student involvement in work sponsored by a company in which the faculty mentor had a financial interest. CONCLUSIONS: Most policies on conflict of interest in our sample of major research institutions in the United States lack specificity about the kinds of relationships with industry that are permitted or prohibited. Wide variation in management of conflicts of interest among institutions may cause unnecessary confusion among potential industrial partners or competition among universities for corporate sponsorship that could erode academic standards. It is in the long-term interest of institutions to develop widely agreed-on, clear, specific, and credible policies on conflicts of interest. JAMA. 2000;284:2203-2208. PMID- 11056592 TI - Assessing faculty financial relationships with industry: A case study. AB - CONTEXT: A growing number of academic researchers receive industry funding for clinical and basic research, but little is known about the personal financial relationships of researchers with their industry sponsors. OBJECTIVES: To assess the extent to which faculty researchers have personal financial relationships with the sponsors of their research, the nature of those financial relationships, and efforts made at the institutional level to address disclosed financial relationships and perceived conflicts of interest. DESIGN AND SETTING: Case study of the University of California, San Francisco (UCSF). Data sources included disclosure forms and official documents maintained by the UCSF Office of Research Administration from December 1980 to October 1999, including decisions made by the UCSF Chancellor's Advisory Panel on Relations with Industry. MAIN OUTCOME MEASURES: Number and types of personal financial relationships with external sponsors (positive financial disclosures from all clinical, basic, or social science faculty who were principal investigators), amount of annual income received from sponsors, and decisions and management strategies used by the advisory panel. RESULTS: By 1999, almost 7.6% of faculty investigators reported personal financial ties with sponsors of their research. Throughout the study period, 34% of disclosed relationships involved paid speaking engagements (range, $250-$20, 000 per year), 33% involved consulting agreements between researcher and sponsor (range, <$1000-$120,000 per year), and 32% involved the investigator holding a position on a scientific advisory board or board of directors. Fourteen percent involved equity ownership, and 12% involved multiple relationships. The advisory panel recommended managing perceived conflicts of interest in 26% of the cases, including recommending the sale of stock, refusing additional payment for talks, resigning from a management position, or naming a new principal investigator for a project. CONCLUSIONS: Faculty researchers are increasingly involved in financial relationships with their research sponsors. Guidelines for what constitutes a conflict and how the conflict should be managed are needed if researchers are to have consistent standards of behavior among institutions. JAMA. 2000;284:2209-2214. PMID- 11056593 TI - Efficacy and safety of recombinant human nerve growth factor in patients with diabetic polyneuropathy: A randomized controlled trial. rhNGF Clinical Investigator Group. AB - CONTEXT: Nerve growth factor is a neurotrophic factor that promotes the survival of small fiber sensory neurons and sympathetic neurons in the peripheral nervous system. Recombinant human nerve growth factor (rhNGF) has demonstrated efficacy as treatment for peripheral neuropathy in experimental models and phase 2 clinical trials. OBJECTIVE: To evaluate the efficacy and safety of a 12-month regimen of rhNGF in patients with diabetic polyneuropathy. DESIGN: Randomized, double-blind, placebo-controlled phase 3 trial conducted from July 1997 through May 1999. SETTING: Eighty-four outpatient centers throughout the United States. PATIENTS: A total of 1019 men and women aged 18 to 74 years with either type 1 or type 2 diabetes and a sensory polyneuropathy attributable to diabetes. INTERVENTIONS: Patients were randomly assigned to receive either rhNGF, 0.1 microg/kg (n = 504), or placebo (n = 515) by subcutaneous injection 3 times per week for 48 weeks. Patients were assessed at baseline, 12 weeks, 24 weeks, and 48 weeks. MAIN OUTCOME MEASURES: The primary outcome measure was a change in neuropathy between baseline and week 48, demonstrated by the Neuropathy Impairment Score for the Lower Limbs, compared between the 2 groups. Secondary outcome measures included quantitative sensory tests using the CASE IV System, the Neuropathy Symptom and Change questionnaire, the Patient Benefit Questionnaire (PBQ), and a global symptom assessment, as well as nerve conduction studies and occurrence of new plantar foot ulcers. Patients also were evaluated for presence of adverse events. RESULTS: Among patients who received rhNGF, 418 (83%) completed the regimen compared with 461 (90%) who received placebo. Administration of rhNGF was safe, with few adverse events attributed to treatment apart from injection site pain/hyperalgesia and other pain syndromes. However, neither the primary end point (P =.25) nor most of the secondary end points demonstrated a significant benefit of rhNGF. Exceptions were the global symptom assessment (P =.03) and 2 of 32 comparisons within the PBQ, which showed a modest but significant benefit of rhNGF (P =.05 for severity of pain in the legs and P =.003 for 6-month symptoms in the feet and legs). CONCLUSION: Unlike previous phase 2 trials, this phase 3 clinical trial failed to demonstrate a significant beneficial effect of rhNGF on diabetic polyneuropathy. JAMA. 2000;284:2215-2221. PMID- 11056594 TI - Safety of lumbar puncture for children with acute lymphoblastic leukemia and thrombocytopenia. AB - CONTEXT: Patients with thrombocytopenia are at risk for spontaneous or procedure related hemorrhage. Whether such patients can safely undergo lumbar puncture (LP) without prophylactic platelet transfusion is unknown. OBJECTIVE: To determine whether an association exists between thrombocytopenia and LP complications among children with acute lymphoblastic leukemia. DESIGN, SETTING, AND PATIENTS: Retrospective review of the records of 958 consecutive children (median age, 5.5 years) with newly diagnosed acute lymphoblastic leukemia who were treated at a pediatric cancer center between February 1984 and July 1998. INTERVENTIONS: All patients underwent a diagnostic LP followed by a median of 4 LPs to instill intrathecal chemotherapy. MAIN OUTCOME MEASURE: Serious complications of LP occurring during the remission induction and consolidation treatment periods (when thrombocytopenia is likely to occur), defined as any neurologic, infectious, or hemorrhagic problems related to the procedure, reported by platelet count at the time of the procedure. RESULTS: Of the 5223 LPs evaluated, 29 were performed at platelet counts of 10 x 10(9)/L or less, 170 at platelet counts of 11 to 20 x 10(9)/L, and 742 at platelet counts of 21 to 50 x 10(9)/L. No serious complications were encountered, regardless of the platelet count. The 95% confidence interval for the proportion of serious complications in the 199 patients with platelet counts of 20 x 10(9)/L or less was 0% to 1.75% and that for the 941 patients with platelet counts of 50 x 10(9)/L or less was 0% to 0.37%. CONCLUSIONS: In our study of children undergoing remission induction or consolidation therapy for acute lymphoblastic leukemia, serious complications of LP were not observed, regardless of platelet count. Prophylactic platelet transfusion is not necessary in children with platelet counts higher than 10 x 10(9)/L. Due to the small number of patients in our study with platelet counts of 10 x 10(9)/L or less, conclusions cannot yet be drawn for such patients. JAMA. 2000;284:2222-2224. PMID- 11056595 TI - A 47-year-old woman with severe asthma. PMID- 11056596 TI - Conflicts of interest in biomedical research. PMID- 11056597 TI - Conflict of interest and the public trust. PMID- 11056598 TI - Msjama: on the cover PMID- 11056599 TI - MSJAMA: the rise of direct-to-consumer advertising of prescription drugs in the United States. PMID- 11056600 TI - MSJAMA: rethinking the role of the learned intermediary: the effect of direct-to consumer advertising on litigation. PMID- 11056601 TI - MSJAMA: challenges in regulating direct-to-consumer advertising. PMID- 11056602 TI - MSJAMA: gifts to physicians in the consumer marketing era. PMID- 11056604 TI - MSJAMA: it was too soon when I went down PMID- 11056603 TI - MSJAMA: direct-to-consumer advertising of prescription drugs: implications for the patient-physician relationship. PMID- 11056605 TI - MSJAMA: grandpa PMID- 11056606 TI - Creating macroscopic atomic Einstein-Podolsky-Rosen states from Bose-Einstein condensates. AB - We present a scheme for creating quant entangled atomic states through the coherent spin-exchange collision of a spinor Bose-Einstein condensate. The state generated possesses macroscopic Einstein-Podolsky-Rosen correlation and the fluctuation in one of its quasispin components vanishes. We show that an elongated condensate with large aspect ratio is most suitable for creating such a state. PMID- 11056607 TI - Squeezing and entanglement of atomic beams. AB - We propose and analyze a scheme for generating entangled atomic beams out of a Bose-Einstein condensate using spin-exchanging collisions. In particular, we show how to create both atomic squeezed states and entangled states of pairs of atoms. PMID- 11056608 TI - Energy transduction in periodically driven non-hermitian systems. AB - We show a new mechanism to extract energy from nonequilibrium fluctuations typical of periodically driven non-Hermitian systems. The transduction of energy between the driving force and the system is revealed by an anomalous behavior of the susceptibility, leading to a diminution of the dissipated power and consequently to an improvement of the transport properties. The general framework is illustrated by the analysis of some relevant cases. PMID- 11056610 TI - Grand unified theories without the desert. AB - We present a grand unified theory (GUT) that has GUT fields with masses of the order of a TeV, but at the same time preserves (at the one-loop level) the success of gauge-coupling unification of the minimal supersymmetric standard model (MSSM) and the smallness of proton decay operators. This scenario is based on a five-dimensional theory with the extra dimension compactified as in the Randall-Sundrum model. The MSSM gauge sector and its GUT extension live in the 5D bulk, while the matter sector is localized on a 4D boundary.the is a test again PMID- 11056609 TI - Tau neutrinos favored over sterile neutrinos in atmospheric muon neutrino oscillations. AB - The previously published atmospheric neutrino data did not distinguish whether muon neutrinos were oscillating into tau neutrinos or sterile neutrinos, as both hypotheses fit the data. Using data recorded in 1100 live days of the Super Kamiokande detector, we use three complementary data samples to study the difference in zenith angle distribution due to neutral currents and matter effects. We find no evidence favoring sterile neutrinos, and reject the hypothesis at the 99% confidence level. On the other hand, we find that oscillation between muon and tau neutrinos suffices to explain all the results in hand. PMID- 11056611 TI - Phenomenological evidence for gluon depletion in pA collisions. AB - The data of J/psi suppression at large x(F) in pA collisions are used to infer the existence of gluon depletion as the projectile proton traverses the nucleus. The modification of the gluon distribution is studied by use of a convolution equation whose nonperturbative splitting function is determined phenomenologically. The depletion factor at x(1) = 0.8 is found to be about 25% at A = 100. PMID- 11056612 TI - Transverse energy fluctuations and the pattern of J/psi suppression in Pb-Pb collisions. AB - The NA50 Collaboration has recently observed that the J/psi production rate in Pb Pb collisions decreases more rapidly as a function of the transverse energy for the most central collisions than for less central ones. We show that this phenomenon can be understood as an effect of transverse energy fluctuations in central collisions. A good fit of the data is obtained using a model which relates J/psi suppression to the local energy density. Our results suggest that the J/psi is completely suppressed at the highest densities achieved in Pb-Pb collisions. PMID- 11056613 TI - Geometric chaoticity leads to ordered spectra for randomly interacting fermions. AB - A rotationally invariant random interaction ensemble was realized in a single- j fermion model. A statistical approach reveals the random coupling of individual angular momenta as a source for the empirically known dominance of ground states with zero and maximum spin. The interpretation is supported by the structure of the ground state wave functions. PMID- 11056614 TI - Minimization methods for the one-particle dirac equation. AB - Taking into account relativistic effects in quantum chemistry is crucial for accurate computations involving heavy atoms. Standard numerical methods can deal with the problem of variational collapse and the appearance of spurious roots only in special cases. The goal of this Letter is to provide a general and robust method to compute particle bound states of the Dirac equation. PMID- 11056615 TI - Slow photoelectron imaging. AB - Experiments are reported on the detection of slow photoelectrons resulting from the photoionization of Xe atoms in a dc electric field by electron imaging. In the far-field photoelectron velocity distributions we can distinguish between direct and indirect ionization processes (involving long range Coulomb interactions with the Xe+ ion). Also, a new modulation of the velocity distribution is observed which cannot be explained by previously discussed mechanisms. Classical and quantum mechanical calculations are presented to support the interpretations. PMID- 11056616 TI - High-resolution vacuum-ultraviolet spectroscopy of an electron-cooled D- beam. AB - The shape parameters for the lowest-lying (1)P(o) resonance, (2)?0?-3, of D- have been measured using high-resolution vacuum-ultraviolet spectroscopy. The experiment was performed at the storage ring ASTRID, and the resonance was resolved by applying electron cooling to reduce the velocity spread of the ion beam. The resonance has a width of 37(3) microeV while the asymmetry parameter q of the Fano profile is -16(3). These values present the first critical test of a large number of theoretical calculations. PMID- 11056617 TI - Pondermotive forces with slow light. AB - This work describes atomic processes which result from the greatly enhanced longitudinal gradient force which is inherent to the propagation of slow light. These processes are (1) ballistic atom motion and atom surfing, and (2) a type of local pondermotive nonlinearity or scattering which results from free-particle sinusoidal motion and the density variation caused by this motion. PMID- 11056618 TI - Incremental approach to strongly correlated many-body finite systems. AB - The transition and the Green operators of an interacting N body system are obtained from the solutions of the N-M body problem where M = 1,2,ellipsis,N-2. This is achieved via the development of a cumulative, nonperturbative approach that makes use of existing knowledge on the system when the number of interacting particles is reduced. The method is applied to four interacting Coulomb particles where the Green operator is expressed as a sum of Green operators of all three body subsystems that can be combined within the four body system. The calculated four particle continuum spectrum is in a remarkable agreement with recent experimental findings. PMID- 11056619 TI - Angular momentum partitioning and hexacontatetrapole moments in impulsively excited argon ions. AB - We have studied polarized electron collisions with Ar in which the target is simultaneously ionized and excited to form Ar+(3p(4)(1D)4p) states. We measured the integrated Stokes parameters of the subsequent fluorescence emitted by the (2)F(7/2), (2)F(5/2), (2)D(5/2), and (2)P(3/2) states along the direction of electron polarization. The Rubin-Bederson hypothesis is shown to hold for the L and S multipoles of these states. The electric quadrupole and hexadecapole of the 1D core are derived. By recoupling these moments with the electric quadrupole moment of the 4p electron, we calculate higher moments of the total ionic orbital angular momentum, including its hexacontatetrapole (64-pole) moment. PMID- 11056620 TI - Frequency modulation in the transmittivity of wave guides in elastic-wave band gap materials. AB - It is shown, for the first time, that the transmittivity of wave guides created as rectilinear defects in periodic elastic band-gap materials oscillates as a function of frequency. The results are obtained using the finite difference time domain method for elastic waves propagating in two-dimensional inhomogeneous media. The oscillations of the transmittivity are due to the richness of modes in the elastic systems and, mainly, due to the periodicity of the potential in the direction of the wave propagation. Results are presented for a periodic array of Pb and Ag cylinders inserted in an epoxy host, as well as for Hg cylinders in an Al host. PMID- 11056621 TI - Inversion symmetry and exact critical exponents of dissipating waves in the sandpile model. AB - By an inversion symmetry, we show that in the Abelian sandpile model the probability distribution of dissipating waves of topplings that touch the boundary of the system shows a power-law relationship with critical exponent 5/8 and the probability distribution of those dissipating waves that are also last in an avalanche has an exponent of 1. Our extensive numerical simulations not only support these predictions, but also show that inversion symmetry is useful for the analysis of the two-wave probability distributions. PMID- 11056622 TI - Experimental dynamics of a vortex within a vortex. AB - We report the experimental dynamics of a new two-dimensional (2D) fluid phenomenon that occurs when an intense, pointlike vortex is placed within a diffuse, circular vortex. Our observations, made using strongly magnetized electron columns to model the 2D fluid, support the analysis performed by Jin and Dubin. PMID- 11056623 TI - Solitary coherent structures in viscoelastic shear flow: computation and mechanism. AB - Starting from stationary bifurcations in Couette-Dean flow, we compute nontrivial stationary solutions in inertialess viscoelastic circular Couette flow. These solutions are strongly localized vortex pairs, exist at arbitrarily large wavelengths, and show hysteresis in the Weissenberg number, similar to experimentally observed "diwhirl" patterns. Based on the computed velocity and stress fields, we elucidate a heuristic, fully nonlinear mechanism for these flows. We propose that these localized, fully nonlinear structures comprise fundamental building blocks for complex spatiotemporal dynamics in the flow of elastic liquids. PMID- 11056624 TI - NonliNEAR vertical oscillations of a particle in a sheath of a rf discharge. AB - A new simple method to measure the spatial distribution of the electric field in the plasma sheath is proposed. The method is based on the experimental investigation of vertical oscillations of a single particle in the sheath of a low-pressure radio-frequency discharge. It is shown that the oscillations become strongly nonlinear as the amplitude increases. The theory of anharmonic oscillations provides a good quantitative description of the data and gives estimates for the first two anharmonic terms in an expansion of the sheath potential around the particle equilibrium. PMID- 11056625 TI - Three-dimensional strongly coupled plasma crystal under gravity conditions. AB - Experiments were carried out to investigate a three-dimensional (3D) plasma crystal. A method of determining the positions of each individual microparticle has been developed. A crystal volume of about 2x10(4) particles in 19 horizontal planes was analyzed. Direct imaging and the 3D pair correlation function show that "domains" of fcc and hcp lattices coexist in the crystal. Other structures, in particular, the theoretically predicted bcc lattice, were not observed. PMID- 11056626 TI - Stimulated raman scattering of rapidly amplified short laser pulses. AB - The theory of transient forward stimulated Raman scattering (FSRS) of rapidly amplified short laser pulses is put forth to complement the classical theory for FSRS of stationary pulses. Quantitative conditions for FSRS suppression are identified. In particular, it is shown quantitatively how the limitation imposed by pumped pulse FSRS on the output laser intensity in plasma-based ultrapowerful backward Raman amplifiers can be overcome through a selective detuning of the Stokes resonance. PMID- 11056627 TI - Reversible thermal gelation in soft spheres. AB - Upon heating, concentrated solutions of star polymers and block copolymer micelles in a good solvent, representing soft spheres, undergo a reversible gelation. This phenomenon is attributed to the formation of clusters causing a partial dynamic arrest of the swollen interpenetrating spheres at high temperatures. A phase diagram analogous to that of sterically stabilized colloids is proposed. PMID- 11056628 TI - Evidence of two viscous relaxation processes in the collective dynamics of liquid lithium. AB - New inelastic x-ray scattering experiments have been performed on liquid lithium in a wide wave vector range. With respect to the previous measurements, the instrumental resolution, improved up to 1. 5 meV, allows one to accurately investigate the dynamical processes determining the observed shape of the dynamic structure factor S(Q, omega). A detailed analysis of the line shapes shows the coexistence of relaxation processes with both slow and fast characteristic time scales, and therefore shows that pictures of the relaxation mechanisms based on a simple viscoelastic model must be abandoned. PMID- 11056629 TI - Surface plasmon resonance study of the spreading of a liquid-crystal smectic- a droplet on a gold substrate. AB - We report a study of the spreading of liquid-crystal smectic- A droplets which completely wet a gold substrate. After the initial spreading of a molecular precursor layer, the droplets develop a complex terraced structure consisting of several monolayers and one bilayer spreading at different rates. During the late stage of spreading the droplet height decreases via a series of layer-by-layer shrinkages until a final state is reached which consists of a single dense monolayer. PMID- 11056630 TI - Solving the crystallographic phase problem in i(AlPdMn). AB - We apply a new technique for ab initio phase determination [Acta Crystallogr. Sect. A 55, 48 (1999)] to solve for the average structure of the icosahedral ( i) phase of AlPdMn. After an introduction to the crystallographic phase problem and a description of the method, we present a brief report of our findings for the structure of i(AlPdMn). Despite the use of data from extremely high quality samples, we find strong evidence of disorder in the structure, lending support to the random tiling model of quasicrystal stabilization. PMID- 11056631 TI - Dislocated epitaxial islands. AB - Dislocation networks observed in CoSi (2) islands grown epitaxially on Si are compared with the results of dislocation-dynamics calculations. The calculations make use of the fact that image forces play a relatively minor role compared to line tension forces and dislocation-dislocation interactions. Remarkable agreement is achieved, demonstrating that this approach can be applied more generally to study dislocations in other mesostructures. PMID- 11056632 TI - Laser-induced graphitization on a diamond (111) surface. AB - We report an atomistic simulation study of laser-induced graphitization on the diamond (111) surface. Our simulation results show that the diamond to graphite transition occurs along different pathways depending on the length of the laser pulse being used. Under nanosecond or longer laser pulses, graphitization propagates vertically into bulk layers, leading to the formation of diamond graphite interfaces after the laser treatment. By contrast, with femtosecond (0.2 0.5 ps) laser pulses, graphitization of the surface occurs layer by layer, resulting in a clean diamond surface after the ablation. This atomistic picture provides an explanation of recent experimental observations. PMID- 11056633 TI - Critical dynamics of burst instabilities in the Portevin-Le Ch atelier effect. AB - We investigate the Portevin-Le Chatelier effect (PLC), by compressing Al-Mg alloys in a very large deformation range, and interpret the results from the viewpoint of phase transitions and critical phenomena. The system undergoes two dynamical phase transitions between intermittent (or "jerky") and "laminar" plastic dynamic phases. Near these two dynamic critical points, the order parameter 1/tau of the PLC effect exhibits large fluctuations, and "critical slowing down" (i.e., the number tau of bursts, or plastic instabilities, per unit time slows down considerably). PMID- 11056634 TI - Crystal-like high frequency phonons in the amorphous phases of solid water. AB - The high frequency dynamics of low-density (LDA) and high-density (HDA) amorphous ice and of cubic ice ( I(c)) has been measured by inelastic x-ray scattering in the 1-15 nm(-1) momentum transfer ( Q) range. Sharp phononlike excitations are observed, and the longitudinal acoustic branch is identified up to Q = 8 nm(-1) in LDA and I(c) and up to 5 nm(-1) in HDA. The narrow width of these excitations is in sharp contrast to the broad features observed in all amorphous systems studied so far. The "crystal-like" behavior of amorphous ices, therefore, implies a considerable reduction in the number of decay channels available to soundlike excitations which is interpreted as a sign of low local disorder. PMID- 11056635 TI - Efficient Monte Carlo algorithm and high-precision results for percolation. AB - We present a new Monte Carlo algorithm for studying site or bond percolation on any lattice. The algorithm allows us to calculate quantities such as the cluster size distribution or spanning probability over the entire range of site or bond occupation probabilities from zero to one in a single run which takes an amount of time scaling linearly with the number of sites on the lattice. We use our algorithm to determine that the percolation transition occurs at p(c) = 0.592 746 21(13) for site percolation on the square lattice and to provide clear numerical confirmation of the conjectured 4/3-power stretched-exponential tails in the spanning probability functions. PMID- 11056636 TI - Local functional models of critical correlations in thin films. AB - Recent work on local functional theories of critical inhomogeneous fluids and Ising-like magnets has shown them to be a potentially exact, or near exact, description of universal finite-size effects associated with the excess free energy and scaling of one-point functions in critical thin films. This approach is extended to predict the two-point correlation function G in critical thin films with symmetric surface fields in arbitrary dimension d. In d = 2 we show there is exact agreement with the predictions of conformal invariance for the complete spectrum of correlation lengths xi((n)) as well as the detailed position dependence of the asymptotic decay of G. In d = 3 and d>/=4 we present new numerical predictions for the universal finite-size correlation length and scaling functions determining the structure of G across the thin film. Highly accurate analytical closed form expressions for these universal properties are derived in arbitrary dimension. PMID- 11056637 TI - Optical field-induced mass transport in As2S3 chalcogenide glasses. AB - We report the observation of a photorefractivelike nonlinearity responsible for the formation of giant relief modulations in amorphous semiconductor glasses. The photoinduced softening of the matrix, formation of defects with enhanced polarizability, and their drift under the optical field gradient force is believed to be the origin of the mass transport. PMID- 11056638 TI - Roughness evolution of ion sputtered rotating InP surfaces: pattern formation and scaling laws. AB - The topography evolution of simultaneously rotated and Ar (+) ion sputtered InP surfaces was studied using scanning force microscopy. For certain sputter conditions, the formation of a highly regular hexagonal pattern of close-packed mounds was observed with a characteristic spatial wavelength which increases with sputter time t according to lambda approximately t(gamma) with gamma approximately 0.26. Based on the analysis of the dynamic scaling behavior of the surface roughness, the evolution of the surface topography will be discussed within the limits of existing models for surface erosion by ion sputtering. PMID- 11056639 TI - Atomic transport and chemical stability during annealing of ultrathin Al2O3 films on Si. AB - Ultrathin films of Al2O3 deposited on Si were submitted to rapid thermal annealing in vacuum or in oxygen atmosphere, in the temperature range from 600 to 800 degrees C. Nuclear reaction profiling with subnanometric depth resolution evidenced mobility of O, Al, and Si species, and angle-resolved x-ray photoelectron spectroscopy revealed the formation of Si-Al-O compounds in near surface regions, under oxidizing atmosphere at and above 700 degrees C. Under vacuum annealing all species remained essentially immobile. A model is presented based on diffusion-reaction equations capable of explaining the mobilities and reproducing the obtained profiles. PMID- 11056640 TI - Signature of atomic structure in the quantum conductance of gold nanowires. AB - We have used high resolution transmission electron microscopy to determine the structure of gold nanowires generated by mechanical stretching. Just before rupture, the contacts adopt only three possible atomic configurations, whose occurrence probabilities and quantized conductance were subsequently estimated. These predictions have shown a remarkable agreement with conductance measurements from a break junction operating in ultrahigh vacuum, corroborating the derived correlation between nanowire atomic structure and conductance behavior. PMID- 11056641 TI - Adsorption on carbon nanotubes: quantum spin tubes, magnetization plateaus, and conformal symmetry. AB - We formulate the problem of adsorption onto the surface of a carbon nanotube as a lattice gas on a triangular lattice wrapped around a cylinder. This model is equivalent to an XXZ Heisenberg quantum spin tube. We find density plateau structures for armchair, zigzag, and chiral nanotubes. The zigzags are special and have extensive zero temperature entropy plateaus in the classical limit. Quantum effects lift the degeneracy, leaving gapless excitations described by a c = 1 conformal field theory with compactification radius quantized by the tube circumference. PMID- 11056642 TI - Tunneling between two luttinger liquids induced by a driving field. AB - Tunneling between two Luttinger liquids driven by a time-dependent field with a frequency f is investigated using the zero-mode bosonization. We show that inclusion of the zero modes is essential in order to obtain correct results in the limit L(T)/L>>1 ( L is the channel length and L(T) is the thermal length). We find that the tunneling current is quantized in units of "ef" and takes the form I approximately ef SUM(n = 1)(infinity)delta[mu(F)-2pi / LvPlanck's over 2pi(n-1 / 2)]. PMID- 11056643 TI - Analytic environment-dependent tight-binding bond integrals: application to MoSi2. AB - We present the first derivation of explicit analytic expressions for the environmental dependence of the sigma, pi, and delta bond integrals within the orthogonal two-center tight-binding approximation by using the recently developed bond-order potential theory to invert the nonorthogonality matrix. We illustrate the power of this new formalism by showing that it not only captures the transferability of the bond integrals between elemental bcc Mo and Si and binary C11(b) MoSi2 but also predicts the absence of any discontinuity between first and second nearest neighbors for the ddsigma bond integral even though large discontinuities exist for ppsigma, pppi, and ddpi. PMID- 11056644 TI - Absence of residual quasiparticle conductivity in the underdoped cuprate YBa2Cu4O8. AB - We report measurements of the in-plane thermal conductivity kappa of the stoichiometric underdoped cuprate YBa2Cu4O8 (Y124) below 1 K. kappa(T) is shown to follow a simple phononic T3 dependence at the lowest temperature T for both current directions, with a negligible linear quasiparticle contribution. This observation is in marked contrast with behavior reported in optimally doped cuprates, and implies that extended zero-energy (or low-energy) quasiparticles are absent in Y124. PMID- 11056645 TI - Nonlinear voltage dependence of the shot noise in mesoscopic degenerate conductors with strong electron-electron scattering. AB - It is shown that measurements of zero-frequency shot noise can provide information on electron-electron interaction, because the strong interaction results in the nonlinear voltage dependence of the shot noise in metallic wires. This is due to the fact that the Wiedemann-Franz law is no longer valid in the case of considerable electron-electron interaction. The deviations from this law increase the noise power and make it strongly dependent on the ratio of electron electron and electron-impurity scattering rates. PMID- 11056646 TI - Current switch by coherent trapping of electrons in quantum dots. AB - We propose a new transport mechanism through tunnel-coupled quantum dots based on the coherent population trapping effect. Coupling to an excited level by the coherent radiation of two microwaves can lead to an extremely narrow current antiresonance. The effect can be used to determine interdot dephasing rates and is a mechanism for a very sensitive, optically controlled current switch. PMID- 11056647 TI - Enhancement of persistent currents due to confinement in metallic samples. AB - Confinement and surface roughness (SR) effects on the magnitude of the persistent current are analyzed for ballistic bidimensional metallic samples. Depending on the particular geometry, localized border states can show up at half-filling. These border states contribute coherently to the persistent current and its magnitude is enhanced with respect to their value in the absence of confinement. A linear scaling of the typical current I(typ) with the number of conduction channels M is obtained. This result is robust with respect to changes in the relevant lengths of the samples and to the SR. Possible links of our results to experiments are also discussed. PMID- 11056648 TI - Stability of the smectic quantum hall state: a quantitative study. AB - We present an effective elastic theory which quantitatively describes the stripe phase of the two-dimensional electron gas in high Landau levels ( N>/=2). The dynamical matrix is obtained with remarkably high precision using the time dependent Hartree-Fock approximation. A renormalization group analysis shows that at T = 0, as the partial filling factor Deltanu identical withnu- left floornu right floor moves away from 1/2, the anisotropic conducting state may undergo quantum phase transitions: stripes may get pinned along their conducting direction by disorder, or may lock into one another to form a crystal. The transitions should be reflected in the temperature dependence of the dissipative conductivity. PMID- 11056649 TI - Weak charge quantization as an instanton of the interacting sigma model. AB - Coulomb blockade in a quantum dot attached to a diffusive conductor is considered in the framework of the nonlinear sigma model. It is shown that the weak charge quantization on the dot is associated with instanton configurations of the Q field in the conductor. The instantons have a finite action and are replica nonsymmetric. It is argued that such instantons may play a role in the transition regime to the interacting insulator. PMID- 11056650 TI - Meissner-London currents in superconductors with rectangular cross section. AB - Exact analytic solutions are presented for the magnetic moment and screening currents in the Meissner state of superconductor strips with rectangular cross section in a perpendicular magnetic field and/or with transport current. The extension to finite London penetration is achieved by an elegant numerical method which works also for disks. The surface current in the specimen corners diverges as l(-1/3) where l is the distance from the corner. This enhancement reduces the barrier for vortex penetration and should increase the nonlinear Meissner effect in d-wave superconductors. PMID- 11056651 TI - Effects of interlayer coupling on the irreversibility lines of NbN/AlN superconducting multilayers. AB - We have studied the temperature dependence of the in-plane resistivity of NbN/AlN multilayer samples with varying insulating layer thickness in magnetic fields up to 7 T parallel and perpendicular to the films. The upper critical field shows a crossover from 2D to 3D behavior in parallel fields. The irreversibility lines have the form (1-T/T(c))(alpha), where alpha varies from 4 / 3 to 2 with increasing anisotropy. The results are consistent with simultaneous melting and decoupling transitions for the low anisotropy sample, and with melting of decoupled pancakes in the superconducting layers for higher anisotropy samples. PMID- 11056652 TI - Ferromagnetism and colossal magnetoresistance from phase competition. AB - We report a multicomponent theory for the coexistence of charge ordering (CO), and antiferromagnetic (AFM) and ferromagnetic (FM) spin ordering. This kind of state is invoked for manganites by Moreo et al., Science 283, 2034 (1999) and observed in recent experiments. We show that doping an AFM or CO state always generates a FM component. FM, AFM, and CO necessarily coexist in a particle-hole asymmetric system. Melting of large AFM-CO orders by small magnetic fields and colossal magnetoresistance (CMR) arise whenever the CO and AFM order parameters have similar magnitude and momentum structure. Hole doping favors FM metallic states while electron doping favors AFM-CO states, as in CMR manganites. PMID- 11056653 TI - Experimental observation of disorder-driven hysteresis-loop criticality. AB - We have studied the effect of magnetic disorder on the magnetization reversal process in thin Co/CoO films. The antiferromagnetic CoO layer allows a reversible tuning of the magnetic disorder by simple temperature variation. For temperatures above a critical temperature T(c), we observe a discontinuous magnetization reversal, whereas smooth magnetization loops occur for T 50 cmH2O) and survival. RESULTS: In the crossover trial, peak inspiratory pressure was significantly lower using PRVC than with VC (20 cmH2O vs 24 cmH2O, P < 0.0001). No other statistically significant differences were found. CONCLUSIONS: Peak inspiratory pressure was significantly lower during PRVC ventilation than during VC ventilation, and thus PRVC may be superior to VC in certain patients. However, in this small group of patients, we could not demonstrate that PRVC improved outcome. PMID- 11056700 TI - Prehospital point of care testing of blood gases and electrolytes - an evaluation of IRMA. AB - BACKGROUND: This study evaluated the feasibility of blood gas analysis and electrolyte measurements during emergency transport prior to hospital admission. RESULTS: A portable, battery-powered blood analyzer was used on patients in life threatening conditions to determine pH, pCO2, pO2, sodium, potassium and ionized calcium. Arterial blood was used for blood gas analysis and electrolyte measurements. Venous blood was used for electrolyte measurement alone. During the observation period of 4 months, 32 analyses were attempted on 25 patients. Eleven measurements (34%) could not be performed due to technical failure. Overall, 25 samples taken from 21 patients were evaluated. The emergency physicians (all anesthesiologists) considered the knowledge of blood gases and/or electrolytes to be helpful in 72% of cases. This knowledge led to immediate therapeutic consequences in 52% of all cases. After a short training and familiarization session the handling of the device was found to be problem free. CONCLUSIONS: We concluded that knowledge of the patients' pH, pCO2 and pO2 in life threatening situations yields more objective information about oxygenation, carbon dioxide and acid-base regulation than pulse oximetry and/or capnometry alone. Additionally, it enables physicians to correct severe hypokalemia or hypocalcemia in cases of cardiac failure or malignant arrhythmia. PMID- 11056701 TI - The use of cephalad cannulae to monitor jugular venous oxygen content during extracorporeal membrane oxygenation. AB - BACKGROUND: When used during extracorporeal membrane oxygenation (ECMO), jugular venous bulb catheters, known as cephalad cannulae, increase venous drainage, augment circuit flow and decompress cerebral venous pressure. Optimized cerebral oxygen delivery during ECMO may contribute to a reduction in neurological morbidity. This study describes the use of cephalad cannulae and identifies rudimentary data for jugular venous oxygen saturation (JVO2) and arterial to jugular venous oxygen saturation difference (AVDO2) in this patient population. RESULTS: Patients on venoarterial (VA) ECMO displayed higher JVO2 (P < 0.01) and lower AVDO2 (P = 0.01) than patients on venovenous (VV) ECMO (P < 0.01). During VV ECMO, JVO2 was higher and AVDO2 lower when systemic pH was < 7.35 rather than > 7.4 (P = 0.01). During VA ECMO, similar differences in AVDO2 but not in JVO2 were observed at different pH levels (P = 0.01). CONCLUSIONS: Jugular venous saturation and AVDO2 were influenced by systemic pH, ECMO type and patient age. These data provide the foundation for normative values of JVO2 and AVDO2 in neonates and children treated with ECMO. PMID- 11056702 TI - Impact of renal dysfunction on weaning from prolonged mechanical ventilation. AB - BACKGROUND: In the intensive care unit (ICU) setting, the combination of mechanical ventilation and renal replacement therapy (RRT) has been associated with prolonged length of hospital stay, high cost of care and poor outcome. We gathered outcome data on patients who had severe renal dysfunction on transfer to our regional weaning center (RWC) for attempted weaning from prolonged mechanical ventilation (PMV). We screened the admission laboratory values of 1077 patients transferred to our RWC over an 8-year period. We reviewed the medical records of patients with serum creatinine > 2.5 mg/dl. RESULTS: Sixty-three patients met screening criteria and 40 patients were on RRT at the time of transfer. Eighteen patients had begun chronic RRT at least 2 months prior to admission to the transferring hospital for their current illness. Twenty-two patients had RRT initiated at the transferring hospital. Ten patients had RRT initiated at the RWC; eight patients had improvement or resolution of azotemia at our facility. RRT was withheld at patient/family request in five patients with progressive renal failure. None of the 50 patients who received RRT recovered renal function during treatment at our RWC. Intermittent hemodialysis was the standard RRT at the RWC. Duration of mechanical ventilation prior to transfer to the RWC was 49.7 +/- 33.5 days (mean +/- SD).Outcome of weaning attempts in the 63 patients was as follows: 13% weaned, 3% failed to wean and 84% died. These outcomes were significantly worse (P<0.001) than those in the 1014 patients whose admission serum creatinine was 48 h (n = 314) were prospectively evaluated, and the prognostic factors of NP, which have been identified in previous studies, were recorded. RESULTS: Pneumonia was diagnosed in 82 patients. The overall mortality rate was 34% for patients with NP compared to 17% in those without NP. Multivariate analysis selected the following three prognostic factors as being significantly associated with a higher risk of death: the presence of multiple organ failure [odds ratio (OR) 6.71, 95% CI, P < 0.001]; the presence of adult respiratory distress syndrome (ARDS) (OR 3.03, 95% CI, P < 0.01), and simplified acute physiology score (SAPS)> 9(OR 2.89, 95% CI, P < 0.05). CONCLUSIONS: In mechanically-ventilated patients NP does not represent an independent risk factor for mortality. Markers of severity of illness were the strongest predictors for mortality. PMID- 11056706 TI - A prospective study of tracheopulmonary complications associated with the placement of narrow-bore enteral feeding tubes. AB - BACKGROUND: In order to determine the type and incidence of pulmonary complications associated with the placement of narrow-bore enteral feeding tubes we conducted a prospective, descriptive study in the multidisciplinary intensive care unit (ICU) of a university hospital. All patients that had narrow-bore enteral feeding tubes inserted over a 2-year period (1993-1995) were included. The study required no clinical interventions. RESULTS: Seven hundred and forty feeding tubes were inserted during the study period. In 14 cases (2%), the feeding tube was inserted into the tracheopulmonary system. Five patients (0.7%) suffered a major complication, including two (0.3%) who died from complications directly related to the feeding tube placement. All patients had altered consciousness and 13 of the 14 had endotracheal tubes in place. Malposition of the feeding tube was not predictable from clinical signs and auscultation, but was detectable by chest roentgenogram. CONCLUSIONS: Inadvertent insertion of enteral feeding tubes into the tracheopulmonary system during placement is associated with significant morbidity and mortality. Clinical signs at the time of insertion are not useful in identifying feeding tubes which are malpositioned. In the ICU patient, a chest roentgenogram is required after all feeding tube insertions prior to the initiation of enteral feeding. In the high-risk patient, alternatives to blind feeding tube insertion should be considered. PMID- 11056707 TI - Acute respiratory distress syndrome: estimated incidence and mortality rate in a 5 million-person population base. AB - BACKGROUND: Various estimates of the incidence and mortality rate of the acute (adult) respiratory distress syndrome (ARDS) have been published. The studies that led to those estimates were based on relatively small patient populations and employed variable diagnostic identifiers of ARDS. The purpose of this study was to estimate the incidence of ARDS and its mortality rate from a large database to which refined diagnostic criteria were applied. We conducted a retrospective review of all hospital discharges over a 4-year period, using screening criteria designed to select patients with ARDS. Discharges from all acute care hospitals in the state of Maryland were reviewed using a computer database from the Health Services Cost Review Commission (HSCRC). Patients >/= 12 years of age were included. Screening criteria consisted of ICD-9 codes 518.5 and 518.82 cross-referenced with procedural codes for ventilatory support (96.70, 96.71 and 96.72). Data were normalized to the number of cases per 100,000 people. RESULTS: During the 4-year study period there were 2,501,147 hospitalizations. Applying the ICD-9 ARDS criteria yielded lower and upper limits of 159-205, 439 568, 531-694 and 529-720 cases of ARDS for 1992, 1993, 1994 and 1995, respectively. Normalizing for a population of 5 million yields yearly lower and upper limit rates of 3.2-4.2, 8.8-11.4, 10.6-13.8 and 10.5-14.2 cases of ARDS per 100,000 people. Mortality upper and lower limit rates based upon the same duration, admissions and population were 38-49%, 39-52%, 36-47%, and 36-49%, respectively. CONCLUSIONS: The incidence of ARDS in Maryland is in the range of 10-14 cases per 100,000 people. The ARDS mortality rate is 36% to 52%, similar to that calculated in previous studies. PMID- 11056708 TI - High frequency oscillatory ventilation attenuates the activation of alveolar macrophages and neutrophils in lung injury. AB - BACKGROUND: Recent investigations have shown that leukocyte activation is involved in the pathogenesis of ventilator-associated lung injury. This study was designed to investigate whether the inflammatory responses and deterioration of oxygenation in ventilator-associated lung injury are attenuated by high-frequency oscillatory ventilation (HFO). We analyzed the effects of HFO compared with conventional mechanical ventilation (CMV) on the activation of pulmonary macrophages and neutrophils in 10 female rabbits. RESULTS: After surfactant depletion, the rabbits were ventilated by CMV or HFO at the same mean airway pressure. Surfactant-depletion followed by 4 h mechanical ventilation hindered pulmonary oxygenation in both groups. Impairment of oxygenation was less severe in the HFO group than in the CMV group. In the HFO group the infiltration of granulocytes into alveolar spaces occurred more readily than in the CMV group. Compared with CMV, HFO resulted in greater attenuation of beta2-integrin expression, not only on granulocytes, but also on macrophages. CONCLUSIONS: In the surfactant-depleted lung, the activation of leukocytes was attenuated by HFO. Reduced inflammatory response correlated with decreased impairment of oxygenation. HFO may reduce lung injury via the attenuation of pulmonary inflammation. PMID- 11056709 TI - Effects of an angiotensin II antagonist on organ perfusion during the post resuscitation phase in pigs. AB - BACKGROUND: The aim of this study was to compare pre-arrest and post resuscitation organ perfusion values and to investigate whether, during the post resuscitation phase, administration of the angiotensin II antagonist telmisartan (TELM) 10 min after restoration of spontaneous circulation (ROSC) could improve organ flow in comparison to placebo. RESULTS: Five minutes after ROSC in the TELM group, blood flow in the cortex and myocardium increased to 583% (P < 0.05) and 137% (not significant), respectively, whereas blood flow of the colon, stomach and pancreas decreased to 50% (P < 0.05), 28% (P < 0.05) and 19% (P < 0.05) of pre-arrest values, respectively. At 90 min after ROSC, pre-arrest perfusion values both in non-splanchnic and splanchnic organs were achieved. At no point in time were there significant differences between the two groups with respect to organ blood flow or speed of recovery of organ perfusion. CONCLUSIONS: During the post-resuscitation phase, organ blood flow is characterized by the coincidence of increased cerebral and myocardial blood flow and decreased intestinal blood flow. Administration of TELM 10 min after ROSC did not improve the recovery of organ perfusion. PMID- 11056710 TI - Venous oxygen measurements in the inferior vena cava in neonates with respiratory failure. AB - BACKGROUND: The present study was undertaken to examine the feasibility of venous oxygen measurements in the inferior vena cava (IVC) via a catheter through the umbilical vein. This may serve as a proxy for mixed venous oxygenation and the complications of right atrial cannulation can be avoided at the same time. It has the added advantage of not being affected by atrial right-left shunting. RESULTS: The study included 22 neonates requiring mechanical ventilation for respiratory insufficiency. The success rate of catheterization of the IVC via the umbilical vein was 81% and there was no catheter-related complications. Fifty paired blood samples were obtained and analyzed while the patients were hemodynamically stable. Linear regression analysis showed a poor correlation between arterial oxygen tension (PaO2) and the arterial-venous oxygen content difference [C(a v)O2], r = -0.005, and between PaO2 and the fractional oxygen extraction (FOE), r = -0.114. There was also a poor correlation between arterial oxygen saturation (SaO2) and C(a-v)O2, r = -0.057, and between SaO2 and FOE, r =-0.139. The correlations between venous oxygen tension (PvO2) and C(a-v)O2 and between PvO2 and FOE were r = -0.528 and r = 0.592, respectively. There were good correlations between various oxygen saturation (SvO2) and C(a-v)O2, r = -0.634, and between SvO2 FOE, r = -0.712. CONCLUSION: Venous oxygen measurement in the IVC via an umbilical vein catheter is a simple and safe procedure and provides information about the tissue oxygenation status of critically ill neonates. PMID- 11056711 TI - Sepsis-related organ failure assessment and withholding or withdrawing life support from critically ill patients. AB - BACKGROUND: We studied the incidence of withholding or withdrawing therapeutic measures in intensive care unit (ICU) patients, as well as the possible implications of sepsis-related organ failure assessment (SOFA) in the decision making process and the ethical conflicts emerging from these measures. METHODS: The patients (n = 372) were placed in different groups: those surviving 1 year after ICU admission (S; n = 301), deaths at home (DH; n = 2), deaths in the hospital after ICU discharge (DIH; n = 13) and deaths in the ICU (DI; n = 56). The last group was divided into the following subgroups: two cardiovascular deaths (CVD), 20 brain deaths (BD), 25 deaths after withholding of life support (DWH) and nine deaths after withdrawal of life support (DWD). RESULTS: APACHE III, daily therapeutic intervention scoring system (TISS) and daily SOFA scores were good mortality predictors. The length of ICU stay in DIH (20 days) and in DWH (14 days) was significantly greater than in BD (5 days) or in S (7 days). The number of days with a maximum SOFA score was greater in DWD (5 days) than in S, BD or DWH (2 days). CONCLUSIONS: Daily SOFA is a useful parameter when the decision to withhold or withdraw treatment has to be considered, especially if the established measures do not improve the clinical condition of the patient. Although making decisions based on the use of severity parameters may cause ethical problems, it may reduce the anxiety level. Additionally, it may help when considering the need for extraordinary measures or new investigative protocols for better management of resources. PMID- 11056712 TI - Do pediatric intensivists and radiologists concur on the interpretation of chest radiographs? AB - BACKGROUND: Therapeutic decisions in the pediatric intensive care unit are made by pediatric intensivists (PI) based on their interpretation of chest radiographs before the formal interpretation by a pediatric radiologist (PR). This study was designed to determine the adequacy of chest radiograph interpretations by pediatric intensivists and the effects on patient care. The PI recorded their chest radiograph interpretations, documenting support devices and thoracic abnormalities. Concordance and discordance were determined by the pediatric pulmonologist who was not involved in the care of the patient by comparing the interpretations of the PI and PR. Clinically significant discordance was defined as interpretations by the radiologist that differed to those from the PI that may have required therapeutic intervention. RESULTS: The evaluation of 291 chest radiographs demonstrated an overall concordance rate of 82.5% (240 out of 291; P < 0.05). There was no significant difference in the ability of critical care medicine physicians to identify atelectasis, infiltrates, pleural effusions, or airleaks (P > 0.05). Support devices were correctly identified in 100% of the cases. Discordant interpretations included 20 that were clinically significant, 17 insignificant findings and 14 films over-interpreted by the PI. A chart review of the patients with discordant findings revealed only one finding that required an alteration in therapy. CONCLUSIONS: These findings demonstrate significant agreement between the interpretation of chest radiographs by PI and PR in selected clinical situations. These data support the current practice of the PI making therapeutic decisions based on their interpretations of chest radiographs. PMID- 11056713 TI - A comparison of handwritten and computer-assisted prescriptions in an intensive care unit. AB - BACKGROUND: We conducted a prospective comparative study to evaluate the potential benefit of computer-assisted prescribing (CAP). We compared the accuracy, completeness and time use of CAP with that of conventional handwritten prescribing at the intensive care unit (ICU) of the John Radcliffe Hospital, Oxford, UK. RESULTS: Twenty-five clinicians and 2409 drug entries were evaluated for accuracy, completeness, legibility and time spent prescribing. One hundred and twenty-eight handwritten and 110 CAP charts were monitored. One hundred percent of CAP charts were complete compared to 47% of handwritten charts.Drug prescriptions were divided into three categories: intravenous fluids, intravenous infusions and intermittent drugs. Percentage of correct entries in each category were 64%, 47.5% and 90% for handwritten, compared to 48%, 32% and 90% for CAP charts, respectively.The mean time taken to prescribe was 20 s for hand written prescribing and 55 s for CAP. CONCLUSIONS: Computer-assisted prescriptions were more complete, signed and dated than handwritten prescriptions. Errors in prescribing, including failure to discontinue a drug were not reduced by CAP. Handwritten prescribing was quicker than CAP. Simple enhancements of the computer software could be introduced which might overcome these deficiencies. CAP was successfully integrated into clinical practice in the ICU. PMID- 11056714 TI - Letter to the Editor. PMID- 11056715 TI - Fluctuations of inspired concentrations of nitric oxide and nitrogen dioxide during mechanical ventilation. AB - BACKGROUND: Nitric oxide (NO) is a very reactive agent with potentially toxic oxidation products such as nitrogen dioxide (NO2). Therefore, during NO inhalation a constant inspired concentration and accurate measurement of NO and NO2 concentrations are essential. The objective of this study was to test the NO concentrations at various positions along the inspiratory limb of the breathing circuit using a recently developed system to administer NO in phase with inspiratory flow during mechanical ventilation (Servo 300 NO-A, Siemens, Sweden). Furthermore, we tested whether an active heating system would interfere with inspired NO concentrations. RESULTS: A sharp decline in the NO concentration was found between the respirator's inspiratory outlet and more distal points along the inspiratory limb of the circuit. This finding was most evident when an active heating system was mounted between those points. CONCLUSIONS: The concentrations of NO and NO2 should be measured as near to the patient as possible, as significant fluctuations of these concentrations might be found along the inspiratory limb of the respiratory circuit especially when an active heating system is used. PMID- 11056716 TI - Inhaled nitric oxide in persistent pulmonary hypertension of the newborn refractory to high-frequency ventilation. AB - BACKGROUND: This study was designed to evaluate the effect of nitric oxide (NO) on the management of neonates with severe persistent pulmonary hypertension refractory to high-frequency oscillatory ventilation. METHODS: The birth weight and the gestational age of infants were 3125.5 +/- 794 g (mean +/- SD) and 39 +/- 2.4 weeks, respectively. All neonates were ventilated for an average of 137.5 min (range 90-180 min) prior to NO therapy. The mean oxygenation index (OI) of all neonates prior to NO was 46.3 +/- 5 (mean +/- SEM). NO was initially administered at 20 parts per million (ppm) for at least 2 h and increased gradually by 2 ppm to a maximum of 80 ppm. RESULTS: Eighteen infants (75%) responded and six (25%) did not respond to the treatment. Three neonates died in the responding group, while all the non-responders died (P = 0.0001). The survival rate was 62.5% among all neonates. NO significantly decreased OI (P < 0.0001) and improved the arterial/alveolar (a/A) oxygen ratio (P < 0.0001) within the first 2 h of NO therapy in 61.1% of the responders. However, the OI and the a/A oxygen ratio remained almost the same throughout the treatment in the non-responders and the non-survivors. CONCLUSION: Inhaled NO at 20 ppm, following adequate ventilation for 2 h without significant response, could be used to identify the majority of the non-responders in order to seek other alternatives. PMID- 11056717 TI - Elevated calcitonin precursor levels are related to mortality in an animal model of sepsis. AB - BACKGROUND: Increased serum levels of procalcitonin (ProCT) and its component peptides have been reported in humans with sepsis. Using a hamster model of bacterial peritonitis, we investigated whether serum ProCT levels are elevated and correlate with mortality and hypocalcemia. RESULTS: Incremental increases in doses of bacteria resulted in proportional increases in 72h mortality rates (0, 20, 70, and 100%) as well as increases in serum total immunoreactive calcitonin (iCT) levels at 12 h (250, 380, 1960, and 4020 pg/ml, respectively, vs control levels of 21 pg/ml). Gel filtration studies revealed that ProCT was the predominant (> 90%) molecular form of serum iCT secreted. In the metabolic experiments, total iCT peaked at 12 h concurrent with the maximal decrease in serum calcium. CONCLUSIONS: In this animal model, hyper-procalcitoninemia was an early systemic marker of sepsis which correlated closely with mortality and had an inverse correlation with serum calcium levels. PMID- 11056718 TI - Editorial comment. PMID- 11056719 TI - Saline lavage with substitution of bovine surfactant in term neonates with meconium aspiration syndrome (MAS) transferred for extracorporeal membrane oxygenation (ECMO): a pilot study. AB - BACKGROUND: Meconium aspiration syndrome (MAS) is still a condition associated with a high mortality, and many patients require extracorporeal membrane oxygenation (ECMO) as rescue therapy. Beneficial effects of surfactant and perflubron lavage have been reported. However, pure surfactant supplementation has not been proven to be beneficial in the most severe forms of MAS. This study was performed to demonstrate an improvement in oxygenation in neonates transferred for ECMO and fulfilling ECMO criteria with a saline lavage and surfactant resupplementation. METHODS: Twelve newborns with MAS [gestational age 36-40 weeks, mean birth weight 3200 g, age 4-16 h, oxygenation index (OI) > 40] transferred for ECMO therapy were treated with saline lavage (5-10 cm3/kg body weight, as long as green colored retrieval was observed) and resupplementation with bovine surfactant (Alveofact, Boehringer, Ingelheim, Germany). The OI at admission and 3 h after this procedure was compared using the t-test for paired samples. ECMO was available as rescue therapy at all times. RESULTS: The OI decreased from 49.4 (SD +/- 13.3) to 27.4 (SD +/- 7.3), P < 0.01. The decrease was sustained in nine patients, three patients required ECMO and all patients survived. CONCLUSIONS: As MAS is a condition with parenchymal damage, pulmonary hypertension and obstructive airway disease, no simple causative therapy is possible. Surfactant application after removal of meconium by extensive lavage is feasible as long as 16 h after birth even in infants considered for ECMO therapy; it might reduce the necessity of ECMO. PMID- 11056720 TI - Mechanical ventilation in rural ICUs. AB - BACKGROUND: In recent years, rural hospitals have expanded their scope of specialized services, which has led to the development and staffing of rural intensive care units (ICUs). There is little information about the breadth, quality or outcomes of these services. This is particularly true for specialized ICU services such as mechanical ventilation, where little, if any, information exists specifically for rural hospitals. The long-term objectives of this project were to evaluate the quality of medical care provided to mechanically ventilated patients in rural ICUs and to improve patient care through an educational intervention. This paper reports baseline data on patient and hospital characteristics for both rural and rural referral hospitals. RESULTS: Twenty Iowa hospitals were evaluated. Data collected on 224 patients demonstrated a mean age of 70 years and a mean ICU admission Acute Physiology and Chronic Health Evaluation (APACHE) II score of 22, with an associated 36% mortality. Mean length of ICU stay was 10 days, with 7.7 ventilated days. Significant differences were found in both institutional and patient variables between rural referral hospitals and rural hospitals with more limited resources. A subgroup of patients with diagnoses associated with complex ventilation had higher mortality rates than patients without these conditions. Patients who developed nosocomial events had longer mean ventilator and ICU days than patients without nosocomial events. This study also found ICU practices that frequently fell outside the guidelines recommended by a task force describing minimum standards of care for critically ill patients with acute respiratory failure on mechanical ventilation. CONCLUSIONS: Despite distinct differences in the available resources between rural referral and rural hospitals, overall mortality rates of ventilated patients are similar. Considering the higher mortality rates observed in patients with complicated medical conditions requiring complex ventilation management, the data may suggest that this subgroup could benefit from treatment at a tertiary center with greater resources and technology. PMID- 11056721 TI - The effect of continuous positive airway pressure (CPAP) on renal vascular resistance: the influence of renal denervation. AB - OBJECTIVE: To non-invasively study the effects of continuous positive airway pressure breathing (CPAP) on renal vascular resistance in normal subjects and renal allograft recipients, in other words those with with denervated kidneys. We could then ascertain the influence of renal innervation on any resulting changes in renal haemodynamics. METHODS: Ten healthy volunteers and six renal transplant patients were studied. Using Doppler ultrasonography, the pulsatility index (PI), an index of renovascular resistance, was measured at incremental levels of CPAP (0, 2.5, 5.0 and 7.5 cmH2O). RESULTS: In both groups, the PI increased significantly between 0 and5.0 cmH2O CPAP, with a further increase at 7.5 cmH2O CPAP. CONCLUSIONS: We found that CPAP at 5.0 and 7.5 cmH2O caused a significant increase in renovascular resistance in both normal and renal transplant patients. There was no difference in the degree of rise in renovascular resistance between both groups, indicating that the renal nerves do not play a role in altering renal vascular resistance with the application of CPAP. PMID- 11056722 TI - Cisapride decreases gastric content aspiration in mechanically ventilated patients. AB - OBJECTIVE: To determine the effect of the prokinetic agent cisapride in the prevention of aspiration of gastric contents. DESIGN: A prospective randomized two-period crossover study. SETTING: Fourteen-bed polyvalent intensive care unit in a University Hospital. PATIENTS: Eighteen intubated, mechanically ventilated patients who were seated in a semirecumbent position were studied. METHOD: Tc-99 m sulfur colloid (80 megabecquerels) was administered via nasogastric tube on 2 consecutive days. Patients randomly received cisapride (10 mg, via nasogastric tube) one day and a placebo the other. Bronchial secretions were obtained before and for 5 consecutive h after Tc-99 m administration. The radioactivity was measured in a standard amount (1ml) of bronchial fluid using a gamma counter and expressed as counts per min (cpm) after correction for decay. RESULTS: Sixteen out of 18 (88%) patients had increased radioactivity in bronchial secretions. The radioactivity increased over time both with and without cisapride, although it was lower in patients receiving cisapride than in those receiving a placebo. The cumulative bronchial secretion radioactivity obtained when patients received cisapride was significantly lower than when patients received a placebo: 7540 +/- 5330 and 21965 +/- 16080 cpm, respectively (P <0.05). CONCLUSION: Our results suggest that aspiration of gastric contents exists even in patients who are kept in a semirecumbent position. Moreover, cisapride decreases the amount of gastric contents aspiration in intubated and mechanically ventilated patients and may play a role in the prevention of ventilator associated pneumonia. Cisapride, even with the patient in the semirecumbent position, did not completely prevent gastric content aspiration. PMID- 11056723 TI - Comparison of procalcitonin (PCT) and C-reactive protein (CRP) plasma concentrations at different SOFA scores during the course of sepsis and MODS. AB - OBJECTIVES: The relation of procalcitonin (PCT) plasma concentrations compared with C-reactive protein (CRP) was analyzed in patients with different severity of multiple organ dysfunction syndrome (MODS) and systemic inflammation. PATIENTS AND METHODS: PCT, CRP, the sepsis-related organ failure assessment (SOFA) score, the Acute Physiology, Age, Chronic Health Evaluation (APACHE) II score and survival were evaluated in 40 patients with systemic inflammation and consecutive MODS over a period of 15 days. RESULTS: Higher SOFA score levels were associated with significantly higher PCT plasma concentrations (SOFA 7-12: PCT 2.62 ng/ml, SOFA 19-24: PCT 15.22 ng/ml) (median), whereas CRP was elevated irrespective of the scores observed (SOFT 7-12: CRP 131 mg/l, SOFT 19-24: CRP 135 mg/l). PCT of non-surviving patients was initially not different from that of survivors but significantly increased after the fourth day following onset of the disease, whereas CRP was not different between both groups throughout the whole observation period. CONCLUSIONS: Measurement of PCT concentrations during multiple organ dysfunction syndrome provides more information about the severity and the course of the disease than that of CRP. Regarding the strong association of PCT and the respective score systems in future studies we recommend evaluation also of the severity of inflammation and MODS when PCT concentrations were compared between different types of disease. PMID- 11056724 TI - In-vivo evaluation of simultaneous administration of incompatible drugs in a central venous catheter with a decreased port to port distance. AB - BACKGROUND: Multilumen catheters are commonly used in critically ill children. Their use, however, is associated with significant morbidity. We studied the simultaneous administration of incompatible drugs using a new triple-lumen catheter with decreased length and port to port distances. METHODS: Ten domestic swine, 10-20 kg in weight, were divided into two groups of five. Total parenteral nutrition was administered through the distal port and phenytoin was administered as a bolus and as an infusion in each group. Samples were taken from two sites during the bolus and at 1, 5, and 15 min during phenytoin infusion. Histograms were generated for particle size and concentration. Samples were also examined under the microscope for particles. RESULTS: Histograms of particle size did not show any alteration of the histogram that would suggest particle size > 2 um in diameter in the study or control samples. No particles were identified by phase microscope, light microscope, or Wright stain smear. CONCLUSIONS: The use of a triple-lumen catheter with a distance of 0.4cm between the proximal port and the medial port and 1.3 cm between the medial port and the distal port, for the in vivo simultaneous administration of incompatible solutions does not result in precipitates large enough to cause adverse clinical effects. PMID- 11056725 TI - Variation in red cell transfusion practice in the intensive care unit: a multicentre cohort study. AB - OBJECTIVES: To determine the degree of interinstitutional transfusion practice variation and reasons why red cells are administered in critically ill patients. STUDY DESIGN: Multicentre cohort study combined with a cross-sectional survey of physicians requesting red cell transfusions for patients in the cohort. STUDY POPULATION: The cohort included 5298 consecutive patients admitted to six tertiary level intensive care units in addition to administering a survey to 223 physicians requesting red cell transfusions in these units. MEASUREMENTS: Haemoglobin concentrations were collected, along with the number and reasons for red cell transfusions plus demographic, diagnostic, disease severity (APACHE II score), intensive care unit (ICU) mortality and lengths of stay in the ICU. RESULTS: Twenty five per cent of the critically ill patients in the cohort study received red cell transfusions. The overall number of transfusions per patient day in the ICU averaged 0.95 +/- 1.39 and ranged from 0.82 +/- 1.69 to 1.08 +/- 1.27 between institutions (P < 0.001). Independent predictors of transfusion thresholds (pre-transfusion haemoglobin concentrations) included patient age, admission APACHE II score and the institution (P < 0.0001). A very significant institution effect (P < 0.0001) persisted even after multivariate adjustments for age, APACHE II score and within four diagnostic categories (cardiovascular disease, respiratory failure, major surgery and trauma) (P < 0.0001). The evaluation of transfusion practice using the bedside survey documented that 35% (202 of 576) of pre-transfusion haemoglobin concentrations were in the range of 95-105 g/l and 80% of the orders were for two packed cell units. The most frequent reasons for administering red cells were acute bleeding (35%) and the augmentation of O2 delivery (25%). CONCLUSIONS: There is significant institutional variation in critical care transfusion practice, many intensivists adhering to a 100g/l threshold, and opting to administer multiple units despite published guidelines to the contrary. There is a need for prospective studies to define optimal practice in the critically ill. PMID- 11056726 TI - Heliox improves pulmonary mechanics in a pediatric porcine model of induced severe bronchospasm and independent lung mechanical ventilation. AB - BACKGROUND: A helium-oxygen gas mixture (heliox) has low gas density and low turbulence and resistance through narrowed airways. The effects of heliox on pulmonary mechanics following severe methacholine-induced bronchospasm were investigated and compared to those of a nitrogen-oxygen gas mixture (nitrox) in an innovative pediatric porcine, independent lung, mechanical ventilation model. RESULTS: All of the lungs showed evidence of severe bronchospasm after methacholine challenge. Prospective definition of 'heliox response' was a 15% or greater improvement in lung function in the lung receiving heliox compared with the matched lung receiving nitrox. Seven out of 10 pigs responded to heliox therapy with respect to resistance and eight out of 10 pigs responded to heliox therapy with respect to compliance and tidal volume (P < 0.03). After crossover from nitrox to heliox, eight out of eight lungs significantly improved with respect to tidal volume, resistance and compliance (P < 0.001). After crossover from heliox to nitrox all eight lungs showed a significant increase in resistance and a significant decrease in tidal volume (P < 0.001). CONCLUSIONS: In a pediatric porcine model of acute, severe methacholine-induced bronchospasm and independent lung mechanical ventilation, administration of heliox improves pulmonary mechanics, gas flow, and ventilation. Administration of heliox should be considered for support of pediatric patients with acute, severe bronchospasm requiring mechanical ventilation through small artificial airways. PMID- 11056727 TI - Low systemic vascular resistance: differential diagnosis and outcome. AB - OBJECTIVE: To determine the frequency and prognosis of the various causes of low systemic vascular resistance (SVR). DESIGN: Analysis of consecutive patients over a 5-year period; retrospective review. SETTING: Medical intensive care unit of a large university hospital. PATIENTS: Fifty-five patients with unexplained hypotension and a SVR less than 800 dynes x s/cm5. BACKGROUND: There are minimal data in the medical literature determining the frequency or outcome of patients with a low SVR that is unrelated to sepsis or the sepsis syndrome. We retrospectively reviewed and analyzed all hemodynamic data in a large university hospital over a 5-year period to determine the frequency and prognosis of the various causes of low SVR. Fifty-five patients with unexplained hypotension and a SVR less than 800dynesxs/cm5were identified. MAIN RESULTS: Twenty-two patients (Groups 1 and 2) met the criteria for sepsis syndrome. The mean SVR for this group was 445 +/- 168 dynesxs/cm5 with an associated mortality of 50%. Group 3 contained 20 patients with possible sepsis. Thirteen patients (Group 4) were nonseptic. The mean SVR of this group was 435 +/- 180 dynes x s/cm5 with an associated mortality of 46%. Extremely low SVR (below 450 dynes x s/cm5) was associated with a significantly higher mortality regardless of the etiology. CONCLUSIONS: At least a quarter of patients with hypotension and a low SVR have nonseptic etiologies. The patients with nonseptic etiologies have a similar mortality to septic patients. Clinicians should be aware of the wide spectrum of conditions that induce a low SVR. PMID- 11056728 TI - A prospective randomised pilot study of sedation regimens in a general ICU population: a reality-based medicine study. AB - BACKGROUND: For logistical reasons sedation studies are often carried out in elective surgical patients and the results extrapolated to the general intensive care unit (ICU) population. We question the validity of this approach. We compared the two sedation regimens used in our general ICU in a trial structured to mimic clinical practice as closely as possible. RESULTS: Forty patients were randomised to intermittent diazepam or continuous midazolam and sedation monitored with hourly sedation scores; 31 patients completed the study. Scores indicating undersedation were more common with diazepam (P <0.01); overall adequate sedation midazolam 64.7%, diazepam 35.7% (P =0.21). No patient exhibited inappropriately prolonged sedation. Cost was: midazolam AUS$1.98/h; diazepam AUS$0.06/h. CONCLUSION: Both regimens produced rapid onset of acceptable sedation but undersedation appeared more common with the cheaper diazepam regimen. At least 140 patients should be studied to provide evidence applicable to the general ICU population. Used alone, a sedation score may be an inappropriate outcome measure for a sedation trial. PMID- 11056729 TI - Effects on respiratory function of the head-down position and the complete covering of the face by drapes during insertion of the monitoring catheters in the cardiosurgical patient. AB - BACKGROUND: We evaluated the effect on the respiratory gas exchange of the 30 degrees head-down position and the complete covering of the face by sterile drapes. These are used to cannulate the internal jugular vein and position the pulmonary artery catheter in the cardiosurgical patient. During the two manoeuvres, 20 coronary patients and 10 patients with end-stage heart disease were supplied with oxygen (FiO2 =0.4) by a Venturi mask, while 20 coronary patients breathed room air. The arterial blood samples to measure oxygen (PaO2) and carbon dioxide (PaCO2) tension and oxygen saturation (SaO2) were analysed by a blood gas system. RESULTS: The contemporary application of the head-down position and the drapes over the face significantly increased PaO2 and SaO2 in all the patientssupplied with oxygen. Without the head-down position, leaving the drapes over the face, did not significantly change the two parameters in the coronary patients supplied with oxygen, but induced a significant increase in PaO2 and SaO2 in the patients with end-stage heart disease. In the coronary patients that were breathing room air, PaO2 and SaO2 were stable throughout the study. CONCLUSIONS: We conclude that the 30 degrees head-down position and the complete covering of the face by drapes does not interfere with respiratory gas exchange and can be safely performed in coronary patients supplied with oxygen or breathing room air and in patients with end-stage heart disease supplied with oxygen (FiO2 of 0.4). PMID- 11056730 TI - Extracorporeal membrane oxygenation for overwhelming Blastomyces dermatitidis pneumonia. PMID- 11056731 TI - Scanning laser Doppler is a useful technique to assess foot cutaneous perfusion during femoral artery cannulation. AB - BACKGROUND: Measurement of cardiac output and extravascular lung water in critically ill patients using femoral artery double-indicator dilution involves femoral artery catheterization. The potential risk of vascular compromise to the limb may be exacerbated in patients receiving vasopressors. The utility of scanning laser Doppler flowmetry to measure changes in pedal perfusion following catheterization was assessed. RESULTS: There were no significant changes in mean occlusion pressures or in cutaneous perfusion between either leg or between measurement time points, immediately after or 24 h following insertion of the catheters. CONCLUSIONS: Scanning laser Doppler flowmetry is easily used to assess changes in foot perfusion and the effect of interventions that may reduce blood flow to the skin of the foot. Femoral artery catheterization for double-indicator dilution measurements does not reduce calf occlusion pressures or foot skin perfusion in patients receiving vasopressor drugs. PMID- 11056732 TI - A new prototype of an electronic jet-ventilator and its humidification system. AB - BACKGROUND: Adequate humidification in long-term jet ventilation is a critical aspect in terms of clinical safety. AIM: To assess a prototype of an electronic jet-ventilator and its humidification system. METHODS: Forty patients with respiratory insufficiency were randomly allocated to one of four groups. The criterion for inclusion in this study was respiratory insufficiency exhibiting a Murray score above 2. The four groups of patients were ventilated with three different respirators and four different humidification systems. Patients in groups A and B received superimposed high-frequency jet ventilation (SHFJV) by an electronic jet-ventilator either with (group A) or without (group B) an additional humidification system. Patients in group C received high-frequency percussive ventilation (HFPV) by a pneumatic high-frequency respirator, using a hot water humidifier for warming and moistening the inspiration gas. Patients in group D received conventional mechanical ventilation using a standard intensive care unit respirator with a standard humidification system. SHFJV and HFPV were used for a period of 100 h (4days). RESULTS: A significantly low inspiration gas temperature was noted in patients in group B, initially (27.2 +/- 2.5 degrees C) and after 2 days (28.0 +/- 1.6 degrees C) (P < 0.05). The percentage of relative humidity of the inspiration gas in patients in group B was also initially significantly low (69.8 +/- 4.1%; P < 0.05) but rose to an average of 98 +/- 2.8% after 2 h. The average percentage across all four groups amounted to 98 +/- 0.4% after 2 h. Inflammation of the tracheal mucosa was found in patients in group B and the mucosal injury score (MIS) was significantly higher than in all the other groups. Patients in groups A, C and D showed no severe evidence of airway damage, exhibiting adequate values of relative humidity and temperature of the inspired gas. CONCLUSION: The problems of humidification associated with jet ventilation can be fully prevented by using this new jet-ventilator. These data were sustained by nondeteriorating MIS values at the end of the 4-day study period in groups A, C and D. PMID- 11056733 TI - Diagnostic value of gas exchange tests in patients with clinical suspicion of pulmonary embolism. AB - OBJECTIVE: To assess the value of parameters derived from arterial blood gas tests in the diagnosis of pulmonary embolism. METHOD: We measured alveolar arterial partial pressure of oxygen [P(A-a)O2] gradient, PaO2 and arterial partial pressure of carbon diaxide (PaCO2) in 773 consecutive patients with suspected pulmonary embolism who were enrolled in the Prospective Investigative Study of Acute Pulmonary Embolism. DIAGNOSIS: The study design required pulmonary angiography in all patients with abnormal perfusion scans. RESULTS: Of 773 scans, 270 were classified as normal/near-normal and 503 as abnormal. Pulmonary embolism was diagnosed by pulmonary angiography in 312 of 503 patients with abnormal scans. Of 312 patients with pulmonary embolism, 12, 14 and 35% had normal P(A a)O2, PaO2 and PaCO2, respectively. Of 191 patients with abnormal scans and negative angiograms, 11, 13 and 55% had normal P(A-a)O2, PaO2 and PaCO2, respectively. The proportions of patients with normal/near-normal scans who had normal P(A-a)O2, PaO2 and PaCO2 were 20, 25 and 37%, respectively. No differences were observed in the mean values of arterial blood gas data between patients with pulmonary embolism and those who had abnormal scans and negative angiograms. Among the 773 patients with suspected pulmonary embolism, 364 (47%) had prior cardiopulmonary disease. Pulmonary embolism was diagnosed in 151 (41%) of 364 patients with prior cardiopulmonary disease, and in 161 (39%) of 409 patients without prior cardiopulmonary disease. Among patients with pulmonary embolism, there was no difference in arterial blood gas data between patients with and those without prior CPD. CONCLUSION: These data indicate that arterial blood gas tests are of limited value in the diagnostic work-up of pulmonary embolism if they are not interpreted in conjunction with clinical and other laboratory tests. PMID- 11056734 TI - Influence of twenty-five per cent human serum albumin on total and ionized calcium concentrations in vivo. AB - BACKGROUND: A inverse correlation has been found between changes in ionized calcium concentrations and the addition of albumin in vitro, which may explain adverse cardiovascular effects attributed to exogenous albumin in vivo. The purpose of this investigation was to determine the interaction (if any) between exogenous 25% albumin administration (100 ml given over < 30 min) and calcium concentrations in patients, all but one of whom were in an intensive care unit. RESULTS: There were no significant differences in the ionized calcium concentrations obtained before, at the end and 6 h after the administration of albumin (1.09 +/- 0.23, 1.06 +/- 0.22, 1.06 +/- 0.21 mmol/l, respectively). Similarly, there were no significant differences in the total calcium concentrations between these same time periods (2.03 +/- 0.18, 2.05 +/- 0.20, 2.08 +/- 0.23 mmol/l, respectively). CONCLUSIONS: In patients receiving infusions of 25% albumin, it appears that circulating calcium concentrations are well regulated by homeostatic mechanisms. Albumin infusions had no effect on calcium concentrations, although it is possible that temporary changes of questionable clinical importance may have occurred between measurement periods. PMID- 11056735 TI - Compliance characteristics of the Portex Soft Seal Cuff improves seal against leakage of fluid in a pig trachea model. AB - BACKGROUND: The Portex Soft Seal high-volume, low-pressure cuffed tracheal tube was compared with the Mallinckrodt HiLo, Sheridan Preformed and Portex Profile tracheal tubes for leakage of dye placed in the subglottic space of a pig's trachea which was used in a benchtop mechanical ventilation model and in six isolated pig tracheas. RESULTS: There was no leakage, either in the ventilation model or in the isolated tracheas in the Portex Soft Seal group. There was rapid leakage in the ventilation model and in all the isolated tracheas for the Mallinckrodt HiLo, and five out of six isolated tracheas for the Sheridan Preformed and the Portex Profile group. CONCLUSIONS: This benchtop study suggests that the improved compliance characteristics of the Portex Soft Seal cuff are beneficial in preventing leakage of fluid in these models. PMID- 11056736 TI - Polyethylene glycol-superoxide dismutase inhibits lipid peroxidation in hepatic ischemia/reperfusion injury. AB - BACKGROUND: Hepatic injury after ischemia/reperfusion is attributed to the development of oxygen free radical (OFR)-mediated lipid peroxidation--a process that can be measured through its byproducts, specifically malondialdehyde. The use of free radical scavengers can offer significant protection against OFR induced liver injury. We hypothesize that a new potent OFR scavenger, polyethylene glycol-superoxide dismutase (PEG-SOD), can inhibit OFR-mediated lipid peroxidation in hepatic ischemia/reperfusion injury. METHODS: Twelve male Sprague-Dawley rats (300-350 g) were subjected to occlusion of the left and middle hepatic arteries and portal veins for 90 min, followed by 120 min reperfusion. PEG-SOD (5000 units/kg) was given intravenously before vascular occlusion and again immediately upon reperfusion to six rats. Normal saline was given to the remaining six rats to be used as a control group. The right hepatic lobe (used as internal control) and left hepatic lobe were harvested separately and tissue malondialdehyde was measured. RESULTS: A marked increase in lipid peroxide was found in the normal saline group after 2 h reperfusion. Treatment with PEG-SOD prevented the rise in tissue malondialdehyde. The mean difference in the malondialdehyde between the left and right hepatic lobes were 13.20 +/- 6.35 and 1.70 +/- 3.65 nmol/g in the normal saline (control) and PEG-SOD groups, respectively. This difference was found to be statistically significant (P < 0.005) using Student's t-test. CONCLUSIONS: PEG-SOD can effectively attenuate hepatic ischemia/reperfusion injury by inhibiting OFR-mediated lipid peroxidation. PMID- 11056737 TI - Abdominal compartment syndrome: does intra-cystic pressure reflect actual intra abdominal pressure? A prospective study in surgical patients. AB - BACKGROUND: Abdominal compartment syndrome is defined as the adverse physiologic effects of increased intra-abdominal pressure. Prolonged, unrelieved pressure may lead to respiratory compromise, renal impairment, cardiac failure, shock, and death. Abdominal compartment syndrome is diagnosed by measuring intra-cystic pressure as a reflection of intra-abdominal pressure. To examine the validity of the technique, we conducted a prospective study in surgical patients by directly measuring bladder and abdominal pressures simultaneously during laparoscopic cholecystectomy using a previously described technique. RESULTS: In the present model, the bladder had higher baseline pressures than did the abdomen. Measurements across the bladder wall were not identical, but had high positive correlation coefficient when evaluated on an individual basis. Global analysis of the data for all patients showed a weak correlation coefficient. CONCLUSION: In the present study model, intra-cystic pressure did not reflect actual intra abdominal pressure. In spite of some limitations in the study design, we feel that further research is warranted to identify other possible variables that may play a role in the relationship between the urinary bladder and the abdominal cavity pressures, providing better means for diagnosis of abdominal compartment syndrome. PMID- 11056738 TI - The critically ill patient after hepatobiliary surgery. AB - BACKGROUND: We analyzed the causes and results of utilization of critical care services in the special care unit in patients after surgical procedures performed by the hepatobiliary surgical service during a 23-month period. RESULTS: Thirty two of 537 patients (6.0%) required postoperative admission to the special care unit. Twenty-one patients were admitted directly from operating room or from recovery room because of inability to wean from ventilator (n = 10), hypovolemic shock (n = 4), myocardial ischemia or infarction (n = 2), sepsis (n = 2), upper gastrointestinal bleeding (n = 2), and acute renal failure (n =1). Eleven postoperative patients were admitted from floor care for respiratory failure (n = 4), cardiac dysrhythmia or infarction (n = 4), sepsis (n = 2), and upper gastrointestinal bleeding (n = 1). Thirty-eight per cent of patients (n = 12) admitted to the special care unit after surgery died. By multivariate analysis, total postoperative stay in the special care unit that was greater than median total duration of stay of 4.5 days was the only independent predictor of mortality (P = 0.041). CONCLUSIONS: Respiratory failure was the predominant component of all complications after hepatobiliary surgery. No clinically useful predictors of eventual outcome could be identified. PMID- 11056739 TI - Survey of stress ulcer prophylaxis. AB - BACKGROUND: No surveys of stress ulcer prophylaxis prescribing in the USA have been conducted since 1995. Since that time, the most comprehensive meta-analysis and largest randomized study to date concerning stress ulcer prophylaxis have been published. RESULTS: Three hundred sixty-eight surveys were sent to all members of the Section of Pharmacy and Pharmacology of the Society of Critical Care Medicine. One hundred fifty-three (42%) surveys were returned. Representatives from 86% of institutions stated that medications for stress ulcer prophylaxis are used in a majority (>90%) of patients admitted to the intensive care unit (ICU). Twenty-two per cent of institutions have recommendations for both ICU and non-ICU settings. Fifty-eight per cent of institutions stated that there was one preferred medication for stress ulcer prophylaxis, and in 77% of these histamine-2-antagonists were the most popular. CONCLUSIONS: There are wide variations in prescribing practices for stress ulcer prophylaxis. Institutions should consult published literature and use pre-existing guidelines as templates for developing their own guidelines. PMID- 11056740 TI - Thrombocytopenia in critically ill surgical patients: a case-control study evaluating attributable mortality and transfusion requirements. AB - BACKGROUND: That thrombocytopenia results in increased mortality or transfusion requirements has not been confirmed by previous studies. We performed a case control study in which 36 patients who developed severe thrombocytopenia of less than 50x109 platelets/l were carefully matched for the severity of underlying disease and other important variables. RESULTS: Seventeen (47%) thrombocytopenic patients died, versus 10 (28%) matched control patients who were not thrombocytopenic.Nine pairs had a discordant outcome, and in eight of these pairs the thrombocytopenic patient died (exact binomial probability 0.037). The estimated attributable mortality was 19.5% (95% confidence interval 3.2-35.8), and the estimated odds ratio was 2.7 (95% confidence interval 1.02-7.10). Thrombocytopenic patients had comparable values for severity of illness scores between day of admission and day of thrombocytopenia, in contrast with control patients who had a statistically significant decrease in severity of illness scores during the same period. Thirty (83%) of the thrombocytopenic patients required transfusion of blood products, versus 21 (58%) control patients (paired chi2 test 4.92, P < 0.04). The estimated attributable transfusion requirement was 25% (95% confidence interval 5.4-44.6), and the estimated odds ratio was 1.52 (95 confidence interval 1.05-2.20). CONCLUSION: The present study suggests that thrombocytopenia of less than 50 x 109 platelets/l may be a marker for more severe illness and increased risk of death, rather than causative, because a true causal relationship is not established. Thrombocytopenia also leads to an excess of blood product consumption. PMID- 11056741 TI - The immunological effects of continuous veno-venous haemodiafiltration in critically ill patients. AB - BACKGROUND: Haemodynamic instability is common in septic patients with acute renal failure. Continuous veno-venous haemodiafiltration (CVVHD) is therefore used as an alternative to conventional haemodialysis. Haemodialysis is associated with an activation of the immune system. The aim of the present study was to test the hypothesis that initiation of CVVHD influences the immune system with release of proinflammatory cytokines followed by a decrease in granulocyte activation, as assessed by the expression of adhesion molecules. RESULTS: Fifteen patients were included. Mean Acute Physiology and Chronic Health Evaluation-2 score before CVVHD was 19 (range 8-27). Mean duration of CVVHD treatment was 9 days (1-21 days). Tumour necrosis factor-alpha and interleukin-8 were detectable in plasma in all patients, whereas interleukin-10 was detectable only in a few patients. Proinflammatory and anti-inflammatory cytokines were detected in the ultrafiltrate. Large intraindividual and interindividual variations were demonstrated for all of the immunological parameters studied. CONCLUSION: The hypothesis that CVVHD induces the release of proinflammatory cytokines followed by a decrease in granulocyte activation was not confirmed in the present study. The heterogeneous group of patients studied, with different underlying diseases and various durations of illness before the start of CVVHD, might have contributed to the difficulty in demonstrating the proposed immunological effect of CVVHD. PMID- 11056742 TI - Experience with prolonged induced hypothermia in severe head injury. AB - BACKGROUND: Recent prospective controlled trials of induced moderate hypothermia (32-34 degrees C) for relatively short periods (24-48 h) in patients with severe head injury have suggested improvement in intracranial pressure control and outcome. It is possible that increased benefit might be achieved if hypothermia was maintained for more periods longer than 48 h, but there is little in the literature on the effects of prolonged moderate hypothermia in adults with severe head injury. We used moderate induced hypothermia (30-33 degrees C) in 43 patients with severe head injury for prolonged periods (mean 8 days, range 2-19 days). RESULTS: Although nosocomial pneumonia (defined in this study as both new chest radiograph changes and culture of a respiratory pathogen from tracheal aspirate) was quite common (45%), death from sepsis was rare (5%). Other findings included hypokalaemia on induction of hypothermia and a decreasing total white cell and platelet count over 10 days. There were no major cardiac arrhythmias. There was a satisfactory neurological outcome in 20 out of 43 patients (47%). CONCLUSION: Moderate hypothermia may be induced for more prolonged periods, and is a relatively safe and feasible therapeutic option in the treatment of selected patients with severe traumatic brain injury. Thus, further prospective controlled trials using induced hypothermia for longer periods than 48 h are warranted. PMID- 11056743 TI - Balloon laryngoscopy reduces head extension and blade leverage in patients with potential cervical spine injury. AB - BACKGROUND: Head extension and excessive laryngoscope blade levering motion (LBLM) are undesirable during airway management of trauma patients. We hypothesized that laryngoscopy with a modified blade facilitating glottic exposure by balloon inflation would reduce head extension and LBLM. PATIENTS AND METHODS: Seventeen elective surgery patients were enrolled. Patients lay supine with their heads flat on a rigid board and had a rigid collar around their necks. Laryngoscopy was performed with the modified blade and a standard curved blade. Head extension and LBLM angles were determined upon maximal glottic exposure and compared used paired t-tests. Laryngoscopic view grade and oxygen saturation were also determined. RESULTS: Balloon laryngoscopy resulted in less head extension and LBLM (P <0.001). Laryngoscopic view was approximately identical with both blades, and oxygen saturation was always above 97%. CONCLUSIONS: Balloon laryngoscopy reduces head extension and LBLM under simulated cervical spine precautions. PMID- 11056744 TI - Heat stress is associated with decreased lactic acidemia in rat sepsis. AB - BACKGROUND: Elevated plasma lactate has been shown to correlate with mortality in patients with septic shock. Heat stress prior to sepsis has resulted in reduction in acute lung injury and mortality. We investigated whether heat stress resulted in decreased plasma lactate concentration and protected the lung by decreasing the inflammatory response to sepsis. RESULTS: Plasma lactate concentration was elevated in septic rats without prior heat stress. Lactic acid levels were significantly lower in heat-treated septic rats (P < 0.05) and were not significantly different when compared with control rats. Septic rats with or without heat pretreatment had significantly higher myeloperoxidase activity in the lung than did control groups. Heat pretreatment did not prevent neutrophil infiltration or inflammatory mediator production in the lung. CONCLUSION: Prior heat stress ameliorates lactic acidemia in rat sepsis. Heat stress did not attenuate the pulmonary inflammatory process. The mechanism of heat-induced protection from lactic acidemia in sepsis needs to be further explored. PMID- 11056745 TI - Utility of postintubation chest radiographs in the intensive care unit. AB - OBJECTIVE: To determine the clinical usefulness of immediate (stat) chest radiographs after endotracheal intubation when performed by experienced critical care personnel. PATIENTS AND METHODS: This was a prospective study. Endotracheal intubations in an 11-bed intensive care unit and a nine-bed intermediate intensive care unit were included. After intubations were performed by an experienced critical care operator, that individual recorded demographic and procedural data, and predicted radiographic findings on a data collection sheet. Experience at intubation was stratified into four levels of lifetime experience: fewer than 10 procedures, 10-20 procedures, 20-50 procedures, and more than 50 procedures. Radiographic findings evaluated included endotracheal tube position and procedure-related complications. The postintubation chest radiograph was then reviewed and the actual findings were also recorded. RESULTS: A total of 101 evaluable intubations were recorded, two of which were predicted to show tube malposition. Actual radiographic findings revealed 10 malpositions, three of which were too high and seven were too low (one at the level of the carina). A single witnessed aspiration that occurred during intubation was not radiographically apparent until 24 h later. Only the tube positioned at the carina was felt to be of acute clinical significance or to place the patient at any acute risk. CONCLUSIONS: The incidence of endotracheal tube malposition after intubation was underestimated. However, when performed by experienced critical care personnel, acutely significant malpositions were rare (one out of 101 intubations). We conclude that, in the absence of specific pulmonary complications, endotracheal intubations performed by experienced operators may be followed by routine, rather than 'stat' chest radiographs. PMID- 11056746 TI - A new device for 100 per cent humidification of inspired air. AB - STATEMENT OF FINDINGS: A new humidifier for use during mechanical ventilation in endotracheally intubated patients is described and tested. The humidifier is based on a heat-moisture exchanger, which absorbs the expired heat and moisture and releases it into the inspired air. External heat and water are then added at the patient side of the heat-moisture exchanger, so that the inspired gas should reach 100% humidity (44 mg/l) at 37 degrees C. In bench tests using constant and decelerating inspiratory flow and minute volumes of 3-25 l the device gave an absolute humidity of 41-44 mg/l, and it reduced the amount of water consumed in eight mechanically ventilated patients compared with a conventional active humidifier. During a 24-h test period there was no water condensation in the ventilator tubing with the new device. PMID- 11056747 TI - Time-dependency of improvements in arterial oxygenation during partial liquid ventilation in experimental acute respiratory distress syndrome. AB - BACKGROUND: The mechanisms by which partial liquid ventilation (PLV) can improve gas exchange in acute lung injury are still unclear. Therefore, we examined the time- and dose-dependency of the improvements in arterial oxygen tension (PaO2) due to PLV in eight pigs with experimental lung injury, in order to discriminate increases due to oxygen dissolved in perfluorocarbon before its intrapulmonary instillation from a persistent diffusion of the respiratory gas through the liquid column. RESULTS: Application of four sequential doses of perfluorocarbon resulted in a dose-dependent increase in PaO2. Comparison of measurements 5 and 30 min after instillation of each dose revealed a time-dependent decrease in PaO2 for doses that approximated the functional residual capacity of the animals. CONCLUSION: Although oxygen dissolved in perfluorocarbon at the onset of PLV can cause a short-term improvement in arterial oxygenation, diffusion of oxygen through the liquid may not be sufficient to maintain the initially observed increase in PaO2. PMID- 11056748 TI - Femoral vein size in newborns and infants: preliminary investigation. AB - BACKGROUND: The femoral vein is an important site for central venous access in newborns and infants. The objectives of this study are to determine whether age or weight can be used clinically to predict the size of the femoral vein in newborns and infants, and to compare the size of the vein in each individual in both the supine and reverse Trendelenburg positions. RESULTS: Analysis was done in 24 euvolemic individuals, each studied in both the supine and reverse Trendelenburg positions. Twelve of these individuals were newborns and 12 were infants. We used two-factor analysis of variance to explore differences between groups and multiple linear regression analysis to estimate the strength of the relationship between variables. In the infant group, there was a correlation between femoral vein diameter and weight. There was no correlation between weight and vessel size in newborns. In both the newborn and infant groups, vessel diameter increased with subjects in the reverse Trendelenburg position (P < 0.01). CONCLUSION: Weight is predictive of femoral vein diameter in infants, but not in newborns. In infants, weight might serve as a more sensitive index for estimating size of the femoral vein in order to determine accurately the size of intravascular catheter appropriate for cannulation. The diameter of the femoral vein increases in the reverse Trendelenburg position compared with that in the supine position in both newborns and infants. A large prospective study is required to validate these findings. PMID- 11056749 TI - Percutaneous tracheostomy: comparison of Ciaglia and Griggs techniques. AB - BACKGROUND: Although the standard tracheostomy described in 1909 by Jackson has been extensively used in critical patients, a more simple procedure that can be performed at the bedside is needed. Since 1957 several different types of percutaneous tracheostomy technique have been described. The purpose of the present study was to compare two bedside percutaneous tracheostomy techniques: percutaneous dilatational tracheostomy (PDT) and the guidewire dilating forceps (GWDF). MATERIALS AND METHODS: A prospective study in two medical/surgical intensive care units (ICUs) was carried out. Sixty-three critically ill patients who required endotracheal intubation for longer than 15 days were consecutively selected to undergo PDT (25 patients) or GWDF (38 patients) technique. Intraoperative and postoperative complications were recorded. RESULTS: Age (mean +/- standard error) was 63 +/- 1.1 years. The patients had been mechanically ventilated for an average of 19.8 +/- 1.2 days. The GWDF technique was significantly faster than PDT technique (P = 0.02). Fifteen complications occurred in 10 out of 63 (15%) patients. They were as follows: tracheal tear (one patient in each group; in one case this was due to false passage); transient hypotension (one patient in the PDT group and two patients in the GWDF group); atelectasis (one patient in the PDT group); and haemorrhage (one patient in the PDT group and three patients in the GWDF group). In both patients with tracheal tear, reduced arterial oxygen saturation (SaO2) with concomitant subcutaneous emphysema ensued. CONCLUSION: We found no statistical differences between complications with both techniques. The surgical time required for the GWDF technique was less than that for PDT. PMID- 11056750 TI - Hyperbilirubinaemia after major thoracic surgery: comparison between open-heart surgery and oesophagectomy. AB - BACKGROUND: Hyperbilirubinaemia is a common occurrence in patients who are admitted to intensive care units (ICUs) after major surgery, and it is associated with high mortality. We investigated the incidence of hyperbilirubinaemia after two major types of thoracic surgery: open-heart surgery and oesophagectomy. In order to identify the risk factors associated with hyperbilirubinaemia after major surgery, we compared the incidence after open-heart surgery with that after oesophagectomy. RESULTS: Hyperbilirubinaemia was detected in 51% of the open heart surgery patients (n = 133) and in 64% in the oesophagectomy group (n = 74). The incidence of hyperbilirubinaemia was significantly related to the duration of surgery (P< 0.05). In the open-heart surgery group, duration of surgery was 465 +/- 24 min for the patients without hyperbilirubinaemia and 571 +/- 26 min for the patients with hyperbilirubinaemia. In the oesophagectomy group, the procedure durations were 415 +/- 17 min and 493 +/- 20 min, respectively. The overall mortality rate was 8% in the open-heart surgery group; the rate was 12% in those with hyperbilirubinaemia, but 5% in those without hyperbilirubinaemia. No members of the oesophagectomy group died, with or without hyperbilirubinaemia. Infection significantly affected both the occurrence of hyperbilirubinaemia and mortality in the open-heart surgery group. In the subgroups from the open-heart surgery group, 5% (three out of 65) of those without hyperbilirubinaemia (or evidence of infection) died; of the patients with hyperbilirubinaemia, 3% (one out of 38) of those without infection died and 23% (seven out of 30) with detected infection died. CONCLUSION: After open-heart surgery and oesophagectomy, approximately half of the patients studied had higher levels of serum total bilirubin. Time spent in surgery was significantly related to the occurrence of hyperbilirubinaemia. Infection significantly affected mortality and total bilirubin levels after open heart surgery. Control of infection plays a crucial role in the prevention of hyperbilirubinaemia and in reducing mortality. PMID- 11056751 TI - Helium-oxygen mixture does not improve gas exchange in mechanically ventilated children with bronchiolitis. AB - STATEMENT OF FINDINGS: Varying concentrations of helium-oxygen (heliox) mixtures were evaluated in mechanically ventilated children with bronchiolitis. We hypothesized that, with an increase in the helium:oxygen ratio, and therefore a decrease in gas density, ventilation and oxygenation would improve in children with bronchiolitis. Ten patients, aged 1-9 months, were mechanically ventilated in synchronized intermittent mandatory ventilation (SIMV) mode with the following gas mixtures delivered at 15-min intervals: 50%/50% nitrogen/oxygen, 50%/50% heliox, 60%/40% heliox, 70%/30% heliox, and return to 50%/50% nitrogen/oxygen. The use of different heliox mixtures compared with 50%/50% nitrogen/oxygen in mechanically ventilated children with bronchiolitis did not result in a significant or noticeable decrease in ventilation or oxygenation. PMID- 11056752 TI - Relationships between volume and pressure measurements and stroke volume in critically ill patients. AB - OBJECTIVE: To evaluate the relationships between the changes in stroke volume index (SVI), measured in both the aorta and the pulmonary artery, and the changes in intrathoracic blood volume index (ITBVI), as well as the relationship between changes in aortic SVI and changes in the pulmonary artery wedge pressure (PAWP). DESIGN: Prospective study with measurements at predetermined intervals. SETTING: Medical intensive care unit of a university hospital. PATIENTS AND METHODS: One hundred and fifty-four measurements were taken in 45 critically ill patients with varying underlying disorders. Aortic SVI and pulmonary arterial SVI were determined with thermodilution. PAWP was measured using a pulmonary artery catheter. ITBVI was determined with thermal-dye dilution, using a commercially available computer system. RESULTS: A good correlation was found between changes in ITBVI and changes in aortic SVI. However, this correlation weakened when changes in ITBVI were plotted against changes in pulmonary arterial SVI, which was in part probably due to mathematical coupling between ITBVI and aortic SVI. A good correlation between changes in ITBVI and changes in aortic SVI could also be established in most of the individual patients. No correlation was found between changes in PAWP and changes in aortic SVI. CONCLUSION: ITBVI seems to be a better predictor of SVI than PAWP. ITBVI may be more suitable than PAWP for assessing cardiac filling in clinical practice. PMID- 11056753 TI - A case of central nervous system vasculitis related to an episode of Guillain Barre syndrome. AB - The authors report their knowledge about an uncommon case of isolated vasculitis, restricted to the left sylvian artery during an auto-immune Guillain-Barre syndrome (GBS), sustained by cytomegalovirus (CMV). An acute cardiopulmonary failure requiring a ventilator and vasopressor support manifested, notwithstanding plasma exchanging and immune-modulating therapy. An IgM-enriched formula administration coincided with a rapid amelioration of GBS and vasculitis to a complete recovery the next month after her discharge to a rehabilitation centre. PMID- 11056754 TI - Lack of agreement between tonometric and gastric juice partial carbon dioxide tension. AB - STATEMENT OF FINDINGS: Our goal was to compare measurement of tonometered saline and gastric juice partial carbon dioxide tension (PCO2). In this prospective observational study, 112 pairs of measurements were simultaneously obtained under various hemodynamic conditions, in 15 critical care patients. Linear regression analysis showed a significant correlation between the two methods of measuring PCO2 (r(2) = 0.43; P < 0.0001). However, gastric juice PCO2 was systematically higher (mean difference 51 mmHg). The 95% limits of agreement were 315 mmHg and the dispersion increased as the values of PCO2 increased. Tonometric and gastric juice PCO2 cannot be used interchangeably. Gastric juice PCO2 measurement should be interpreted with caution. PMID- 11056755 TI - Quantitation of Pseudomonas aeruginosa in wound biopsy samples: from bacterial culture to rapid 'real-time' polymerase chain reaction. AB - STATEMENT OF FINDINGS: We developed a real-time detection (RTD) polymerase chain reaction (PCR) with rapid thermal cycling to detect and quantify Pseudomonas aeruginosa in wound biopsy samples. This method produced a linear quantitative detection range of 7 logs, with a lower detection limit of 103 colony-forming units (CFU)/g tissue or a few copies per reaction. The time from sample collection to result was less than 1h. RTD-PCR has potential for rapid quantitative detection of pathogens in critical care patients, enabling early and individualized treatment. PMID- 11056756 TI - Respiratory effects of dexmedetomidine in the surgical patient requiring intensive care. AB - STATEMENT OF FINDINGS: The respiratory effects of dexmedetomidine were retrospectively examined in 33 postsurgical patients involved in a randomised, placebo-controlled trial after extubation in the intensive care unit (ICU). Morphine requirements were reduced by over 50% in patients receiving dexmedetomidine. There were no differences in respiratory rates, oxygen saturations, arterial pH and arterial partial carbon dioxide tension (PaCO2) between the groups. Interestingly the arterial partial oxygen tension (PaO2) : fractional inspired oxygen (FIO2) ratios were statistically significantly higher in the dexmedetomidine group. Dexmedetomidine provides important postsurgical analgesia and appears to have no clinically important adverse effects on respiration in the surgical patient who requires intensive care. PMID- 11056757 TI - Spontaneous rupture of malarial spleen: two case reports and review of literature. AB - Malaria has long been among the most common diseases in the southeast Anatolia region of Turkey. In 1992, 18676 cases were diagnosed in Turkey, and Diyarbakir city had the highest incidence (4168 cases), followed by SanliUrfa city (3578 cases). Malaria was especially common during 1994 and 1995, with 84345 and 82094 cases being diagnosed in these years, respectively. Spontaneous rupture of malarial spleen is rare. We saw two cases during 1998, which are reported herein. Both patients were male, and were receiving chloroquine treatment for an acute attack of malaria. One of the patients had developed abdominal pain and palpitations, followed by fainting. The other patient had abdominal pain and fever. Explorative laparotomy revealed an enlarged spleen in both patients. Splenectomy was performed in both patients. We have identified 15 episodes of spontaneous rupture of the spleen in the English language literature published since 1961. Because of increased travel to endemic areas and resistance to antimalarial drugs, malaria is a major medical problem that is becoming increasingly important to surgeons worldwide. Malaria is a particularly important problem in the southeast Anatolia region of Turkey. Prophylactic precautions should be taken by tourists who travel to this region, especially during the summer. PMID- 11056758 TI - Do steroids prevent reintubation in children with laryngotracheobronchitis? AB - BACKGROUND: Classic laryngotrachoebronchitis (LTB) is an inflammatory process, with oedema and secretions that involve the entire laryngotracheobronchial tree. The severity of lower airway disease in African children with LTB has previously been documented. The aim of the present study was to determine whether steroids prevent reintubation in African children with classic LTB. METHOD AND RESULTS: The study was a retrospective analysis from January 1993 to December 1996. Eighty two black children with LTB were mechanically ventilated in the intensive care unit (ICU). By univariate regression, the estimated B coefficients for variables such as age, pneumonia, days of intubation, arterial partial oxygen tension (PaO2) : fractional inspired oxygen (FIO2) ratio, atelectasis and antibiotic use were not statistically significant (P > 0.05) as predictors for reintubation. Using multiple regression (all independent variables in combination), none of the variables acted as predictors of reintubation (P = 0.25). Steroids were shown to have no effect alone or in association with other variables in altering reintubation rates. An increase in the days of intubation showed a tendency towards reintubation (P = 0.06) in the univariate analysis (odds ratio 1.00 1.14), but showed no statistically significant difference in multivariate analysis. Of the variables used as predictors of reintubation, none acted either as a preventive factor or as a risk factor. CONCLUSION: The present results suggest that steroids should not be recommended at any stage in treatment of intubated patients with classic LTB. Prospective studies should evaluate the major risk factors for reintubation: duration of intubation, trauma to the airway at intubation and during ICU stay, and dose and timing of steroids. They should also evaluate whether upper airway disease is present alone or in association with lower airway disease. PMID- 11056759 TI - Prospectively validated predictions of shock and organ failure in individual septic surgical patients: the Systemic Mediator Associated Response Test. AB - INTRODUCTION: Clinically useful predictions of end-organ function and failure in severe sepsis may be possible through analyzing the interactions among demographics, physiologic parameters, standard laboratory tests, and circulating markers of inflammation. The present study evaluated the ability of such a methodology, the Systemic Mediator Associated Response Test (SMART), to predict the clinical course of septic surgery patients from a database of medical and surgical patients with severe sepsis and/or septic shock. PATIENTS AND METHODS: Three hundred and three patients entered into the placebo arm of a multi institutional sepsis study were randomly assigned to a model-building cohort (n = 200; 119 surgical) or to a predictive cohort (n = 103; 55 surgical). Using baseline and baseline plus serial measurements of physiologic data, standard laboratory tests, and plasma levels of IL-6, IL-8, and granulocyte colony stimulating factor (GCSF), multivariate models were developed that predicted the presence or absence of pulmonary edema on chest radiography, and respiratory, renal, coagulation, hepatobiliary, or central nervous system dysfunction and shock in individual patients. Twenty-eight-day survival was predicted also in baseline plus serial data models. These models were validated prospectively by inserting baseline raw data from the 55 surgical patients in the predictive cohort into the models built on the comprehensive training cohort, and calculating the area under the curve (AUC) of predicted versus observed receiver operator characteristic (ROC) plots. RESULTS: SMART predictions of physiologic, respiratory, metabolic, hepatic, renal, and hematologic function indicators were validated prospectively, frequently at clinically useful levels of accuracy. ROC AUC values above 0.700 were achieved in 30 out of 49 (61%) of SMART baseline models in predicting shock and organ failure up to 7 days in advance, and in 30 out of 54 (56%) of baseline plus serial data models. CONCLUSION: SMART multivariate models accurately predict pathophysiology, shock, and organ failure in individual septic surgical patients. These prognostications may facilitate early treatment of end-organ dysfunction in surgical sepsis. PMID- 11056760 TI - Ancient and modern retroviruses. AB - Retroviruses are transmitted in two distinct ways: as infectious particles and as 'endogenous' proviral DNA integrated in the germ line of the host. Modern infectious viruses such as HIV-1 and HIV-2 recently infected mankind from chimpanzee and simian hosts, whereas human endogenous retroviral genomes have been present throughout old world primate evolution. Human T-cell leukemia viruses (HTLV-1 and II) have a much older human provenance than HIV, although new zoonoses from simians may also occur. We have recently characterized new retroviruses in pigs and humans. Porcine endogenous retroviral (PERV) genomes are carried in chromosomal DNA but can be activated to produce virions that are infectious for human cells, which has raised concern over human xenotransplantation using pig tissues. Human retrovirus 5 (HRV-5) is detected as an exogenous genome in association with arthritis and systemic lupus erythematosus. PMID- 11056761 TI - Examination of CD11B/C, CD54 and CD62 expression on cells in trypsin-triggered emphysema model. AB - We studied the CD11b/c, CD54, CD62 expression on BAL (bronco-alveolar lavage) cells of rats by flow cytometry in a trypsin-triggered emphysema model. We made BAL sampling two and a half hour after trypsin infusion, in early inflammatory phase. Rats were divided into three groups: 1. negative controls, 2. saline treated, 3. trypsin-infused rats. We found significantly (p < 0.05) increased number of neutrophil granulocytes in BAL of trypsin-treated group, comparing with controls. By flow cytometry in trypsin-treated group: 1). We found a significantly higher expression of CD54 on BAL macrophages (p < 0.05) 2). There was a lower, not significant CD11b/c expression on neutrophils and on macrophages in BAL, comparing with other groups. 3). A low, but not significant CD62 expression could be detected on neutrophils and on lymphocytes in BAL. We conclude: 1). Two and a half hour after trypsin infusion, macrophages are strongly activated, and play an important role in the neutrophil transendothelial migration in the early inflammatory phase of this model. 2). Neutrophils are high in number in BAL, but they are hardly activated in this early phase. 3). After trypsin infusion having a lower CD62 expression, lymphocytes seem to be involved as well. PMID- 11056762 TI - Relationship between the occurrence of anti-SSA, anti-SSB autoantibodies and HLA class II alleles from the aspect of in vitro inhibitory effect of glucocorticosteroid on the antibody-dependent cellular cytotoxicity in patients with primary Sjogren's syndrome. AB - Though at present there is no evidence-based algorithm for the treatment of primary Sjogren's syndrome, it is generally accepted that glucocorticosteroid (GS) therapy must be introduced in cases with severe systemic manifestations. As the side-effects of the GSs are well known, it would be useful to know in advance how the patients will respond to this type of treatment. For this reason we measured the in vitro steroid sensitivity of 29 SS patients using inhibition of antibody dependent cellular cytotoxicity (ADCC) test by methylprednisolone compared to that of 28 controls. SS patients proved to be significantly less sensitive to GSs than controls (inhibition of ADCC reaction: 42.4 vs 53.1%; p < 0.01). This was especially true in SS patients with anti-SSA and/or SSB autoantibody positivity and with HLA-DR2 and/or -DR3 alleles. Comparing the results of the in vitro GS sensitivity and the clinical effectiveness of the previously applied corticosteroid therapy it seems that steroid inhibition of ADCC reaction has a predictive value in determination of in vivo sensitivity to GSs. However, in patients with decreased in vitro GS sensitivity a more expressed in vivo steroid sensitivity cannot be excluded. PMID- 11056763 TI - The value of pyrolysis mass spectrometry to investigate nosocomial outbreaks caused by Serratia marcescens. AB - Simultaneous outbreaks of S. marcescens infection going on in the Neonatal Intensive Care Unit and the Surgical Department of the same hospital were investigated by pyrolysis mass spectrometry (PyMS). The PyMS analysis of the strains clearly demonstrated that the two outbreaks were caused by different strains. The 14 S. marcescens isolates from the first outbreak were closely related, with the exception of one environmental isolate, which did not harbour the ESBL plasmid, which was present in all other isolates. However, the phage type of all 14 isolates was the same. Among the 9 S. marcescens isolates from the second outbreak, PyMS clearly distinguished 3 that exhibited gentamicin resistance from the remaining 6 gentamicin-susceptible isolates. Phage typing was unhelpful in this case, as none of the isolates were typable. The PyMS typing of nosocomial outbreak strains can reach the level of discrimination approaching that achieved by molecular genetic analysis. PMID- 11056764 TI - Rapid combined assay for Salmonella detection in food samples. AB - A rapid method was developed to detect salmonellae in food samples. The method gave a possibility to obtain results after 28 h 30 min. The preenrichment in buffered peptone water lasted for 6 h, the enrichment in Rappaport-Vassiliadis medium was applied for 18 h followed by PCR with INVA1-INVA2 primer pair, adapting Chiu and Ou's method. This procedure was suitable to demonstrate salmonella contamination at min. 10 cfu/25 g sample. Out of 18 samples there was a good agreement between the results of the conventional and rapid methods in case of 17 samples. PCR with SPVC1-SPVC2 primer pair informing about the presence of virulence plasmid was performed in separate tubes, because decreased sensitivity was observed in case of multiplex PCR. PMID- 11056765 TI - Proteus virulence: involvement of the pore forming alpha-hemolysin (a short review). AB - The genus Proteus belongs to the tribe of Proteae in the family of Enterobacteriaceae, and consists of five species: P. mirabilis, P. vulgaris, P. morganii, P. penneri and P. myxofaciens. They are distinguished from the rest of Enterobacteriaceae by their ability to deaminate phenylalanine and tryptophane. They hydrolyze urea and gelatin and fail to ferment lactose, mannose, dulcitol and malonate; and do not form lysine and arginine decarboxylase or beta galactosidase [1]. Colonies produce distinct "burned chocolate" odor and frequently show the characteristics of swarming motility on solid media. P. mirabilis, P. vulgaris and P. morganii are widely recognized human pathogens. They have been isolated from urinary tract infections, wounds, ear, and nosocomial bacteremic infections, often in immuncompromised patients [2-6]. P. myxofaciens has no clinical interest to this time. P. penneri as species nova was nominated by the recommendation of Hickman and co-workers [7]. Formerly it was recognized as P. vulgaris biogroup 1 or indole negative P. vulgaris [8, 9]. Although it has been less commonly isolated from clinical samples than the other three human pathogenic Proteus species, it has nevertheless been connected with infections of the urinary tract, wounds and has been isolated from the feces of both healthy and diarrheic individuals [10-12]. Potential virulence factors responsible for virulence of Proteae are: IgA protease, urease, type3 fimbriae associated with MR/K haemagglutinins of at least two antigenic types, endotoxin, swarming motility and HlyA and/or HpmA type hemolysins [for review see ref. 13]. In the followings we give a survey of accumulated concepts about the position and characteristics of HlyA type alpha-hemolysins both in general and with emphasis on virulence functions in the tribe of Proteae. PMID- 11056766 TI - Viral contaminants of poliomyelitis vaccines. PMID- 11056767 TI - Orlistat (Xenical) in the management of obesity. PMID- 11056768 TI - Cardiac marker point-of-care testing: evaluation of rapid on-site biochemical marker analysis for diagnosis of acute myocardial infarction. AB - INTRODUCTION: Up to 40% of patients with acute myocardial infarction (AMI) present with non-diagnostic electrocardiograms (ECGs). The diagnosis in such cases is usually made with the aid of biochemical markers. Newer and more rapid assays for such markers have now enabled testing to be done on-site instead of in the laboratory. This potentially enables the clinician to rapidly diagnose and triage patients. We evaluated the diagnostic precision of this point-of-care testing strategy using one such analyser, the Stratus CS (Dade Behring) in a prospective study. MATERIALS AND METHODS: The study population consisted of 51 consecutive patients admitted for suspected AMI with non-diagnostic ECGs. Two blood samples from each patient were drawn simultaneously on admission. The first sample was assayed for myoglobin, troponin I (TnI) and creatine kinase-MB (CKMB) mass by the point-of-care instrument (Stratus CS), and the second sample was sent for standard testing for AMI, comprising a troponin-T (TnT) qualitative test and the analysis of CKMB by the hospital laboratory. Utilising the recommended cut off values for the individual assays, the results of these 2 sets of tests were evaluated based on whether they were positive or negative for AMI and compared against the patient's final diagnosis at discharge. Various combinations of markers were assessed. RESULTS: On evaluation of individual markers, myoglobin was the most sensitive (75%) at 0 to 6 hours after onset of symptoms, while TnI (95%), TnT (80%) and CKMB-mass (90%) performed better at 7 to 12 hours. Point-of care testing utilising a combination of markers was highly sensitive and specific. Both dual-marker panels of myoglobin with TnI and myoglobin with CKMB mass yielded equivalent overall sensitivities and specificities of 90% and 95% respectively. A triple-marker panel of myoglobin, TnI and CKMB-mass had a sensitivity of 93% and specificity of 95%. All point-of-care testing panels had good positive and negative predictive values, and showed comparable diagnostic efficacy with the standard testing presently utilised for the diagnosis of AMI. The average time for results to become available was up to 26 minutes for point of-care testing and 65 minutes for standard testing. CONCLUSION: Point-of-care testing utilising a panel of 2 or 3 cardiac markers has comparable diagnostic precision to the presently utilised testing strategy for AMI, with earlier availability of results. PMID- 11056769 TI - Deep vein thrombosis after total knee replacement. AB - INTRODUCTION: The prevalence of deep vein thrombosis after total knee replacement has been quoted to be between 46% and 84% in the Western literature. The aims of this study were to determine its prevalence in the Singapore population and to assess the need for prophylaxis against deep vein thrombosis. MATERIALS AND METHODS: We examined data on 100 consecutive patients undergoing total knee replacement at the Adult Reconstructive Service, Department of Orthopaedic Surgery, Singapore General Hospital and assessed the possible risk factors: age, sex, weight, previous surgery, unilateral or bilateral surgery, postoperative rehabilitation, tourniquet and operating time. Functional and knee scores based on the Knee Society Clinical Rating System were also assessed. No prophylaxis was given to these patients. These patients underwent a duplex scan of both lower limbs on the seventh postoperative day. Treatment was instituted only if proximal deep vein thrombosis was detected. RESULTS: The overall incidence of deep vein thrombosis was 14% with 64.3% of it occurring distally. Deep vein thrombosis was more common in bilateral total knee replacement (22.2%) compared to unilateral total knee replacement (13.2%). Partial thrombosis was present in 71.4% and occurred predominantly in the ipsilateral leg. There was no evidence of propagation. Only 1 patient developed pulmonary embolism and was treated successfully but there was no evidence of deep vein thrombosis on duplex scan in this patient. CONCLUSION: There was no significant difference in the risk factors between patients who did and those who did not develop deep vein thrombosis. PMID- 11056770 TI - Long-term outcome of aortofemoral bypass for aortoiliac occlusive disease. AB - INTRODUCTION: Revascularisation of aortoiliac occlusive disease has been evolving in the past 2 decades. The present study was undertaken to evaluate the long-term outcomes of aortofemoral bypass for aortoiliac occlusive disease at a tertiary vascular disease centre in Hong Kong. MATERIALS AND METHODS: A retrospective analysis of 94 patients (176 limbs) who survived aortofemoral bypass was performed to evaluate the graft patency, long-term complications, limb loss and patient survival rates. Thirty-six patients were operated for incapacitating claudication (Group I) and 58 for limb salvage (Group II). RESULTS: The overall primary patency rates of aortofemoral bypass were 97%, 90%, 89% and 84% at 1, 3, 5 and 10 years, respectively. Poor distal run-off and neointimal hyperplasia were the leading causes of late graft failure. Other late complications included femoral pseudoaneurysm (n = 1), infection (n = 1) and femoral graft aneurysms (n = 2). The limb loss rate was 5.1% at 4 years. The overall survival rates were 95%, 86%, 81% and 75% at 1, 3, 5 and 10 years, respectively. Ischaemic heart disease and malignancy were the 2 major causes of late death. The 5-year survival rate of group I patients (96%) was significantly superior to that of group II patients (70%). CONCLUSIONS: Aortofemoral bypass achieved a primary patency rate of 89% at 5 years and a satisfactory limb salvage rate. It remains the preferred treatment option for good risk patients with complete occlusion or extensive stenosis of the aortoiliac arteries. PMID- 11056771 TI - Use of low molecular weight heparin for prevention of deep vein thrombosis in total knee arthroplasty--a study of its efficacy in an Asian population. AB - INTRODUCTION: The aims of this paper were to study the incidence of deep vein thrombosis following total knee replacement in an Asian population and to evaluate the role of low molecular weight heparin for deep vein thrombosis in this setting. MATERIALS AND METHODS: We prospectively studied two groups of 100 consecutive patients undergoing total knee replacement separately. Group 1 did not receive any low molecular weight heparin and group 2 received low molecular weight heparin, nodraparin calcium (Fraxiparine) according to body weight. The sex distribution, age group, weight, preoperative knee and function scores, and postoperative rehabilitation were similar for both groups. A single ultrasound technician performed ultrasound duplex scan of both lower limbs on the seventh postoperative day. RESULTS: The incidence of deep vein thrombosis in group 1 was 14% (14 patients, 5 proximal vein thromboses and 9 distal vein thromboses) while in group 2, no patients developed deep vein thrombosis. There was no increased incidence, either local or systemic, of major bleeding complications with the use of low molecular weight heparin. CONCLUSION: While the incidence of deep vein thrombosis following total knee replacement in an Asian population appears lower compared to Western populations, the use of low molecular weight heparin for thromboprophylaxis appears to further reduce the incidence without major bleeding complications. PMID- 11056772 TI - Cardiopulmonary exercise testing in heart transplant candidates. AB - INTRODUCTION: We evaluated the results of patients with congestive heart failure (CHF) who underwent cardiopulmonary exercise testing (CPET) as part of their assessment for heart transplantation in order to examine the relationship between exercise capacity and resting indices of left ventricular function in these patients. MATERIALS AND METHODS: Twenty-seven ambulatory heart transplant candidates underwent CPET using a cycle ergometer and an incremental work-rate protocol till symptom-limitation. These patients included 24 men and 3 women with a mean age of 42.3 years. The aetiology of CHF was coronary artery disease in 14 patients, dilated cardiomyopathy in 11 patients, and congenital heart disease in 2 patients. Mean resting left ventricular ejection fraction (LVEF) was 19% (range 7% to 36%). RESULTS: Thirteen of the 27 tests performed were maximal studies and all except 2 subjects attained a measurable anaerobic threshold during CPET. Of the 13 maximal tests, the causes of exercise limitation were cardiomyopathy in 3 patients, ischaemic heart disease in 2 patients, significant oxygen desaturation in 2 patients, ventilatory limitation due to obstructive lung disease in 1 patient, ventilatory limitation secondary to a restrictive lung disease in 3 patients, and combined obstructive ventilatory and cardiovascular limitation in 2 patients. There was no significant correlation between resting LVEF and peak VO2 percent predicted (r = 0.14, P = 0.49). CONCLUSIONS: Exercise intolerance in patients with CHF may not be related to limited cardiac reserve and non-cardiac causes of exertional symptoms should also be considered. CPET is useful for the evaluation of functional capacity and mechanisms of exercise intolerance in patients with CHF. PMID- 11056773 TI - Prophylactic esmolol infusion for the control of cardiovascular responses to extubation after intracranial surgery. AB - INTRODUCTION: Emergence from general anaesthesia and extubation are often accompanied by significant surges in heart rate and blood pressure. To document these changes and the efficacy of low-dose beta-blocker infusions in ameliorating these rises, we undertook a descriptive dose-ranging study comparing the use of esmolol to placebo in patients emerging from neuro-anaesthesia. MATERIALS AND METHODS: Thirty-six patients undergoing intracranial surgery were randomised to receive saline, esmolol 100 micrograms/kg/min or 200 micrograms/kg/min infusions. The number of patients developing severe hypertension or tachycardia in each group was compared using Fisher's exact test. RESULTS: Systolic blood pressure (SBP) and heart rate (HR) increased in all 3 groups during emergence and peaked at extubation. The proportion of patients with severe tachycardia or hypertension was reduced from 92% in the placebo group to 40% (P = 0.02) and 8% (P = 0.001) in the low and intermediate dose esmolol groups, respectively. Results were better in the intermediate dose group but the difference was not statistically significant. Two patients from the esmolol infusion groups required supplemental medication for bradycardia. CONCLUSION: Severe hypertension or tachycardia occurs in 92% of patients during extubation following neuro-anaesthesia and warrants the consideration of routine prophylaxis. Prophylactic esmolol infusion for the control of haemodynamic disturbances during extubation is feasible and safe. A modest level of obtundation is evident at 100 micrograms/kg/min but a rate of 200 micrograms/kg/min may prove to be more effective. PMID- 11056774 TI - Tracheoesophageal puncture outcomes and predictors of success in laryngectomised patients. AB - INTRODUCTION: Prosthetic voice restoration has been widely used for vocal rehabilitation in laryngectomised patients. The objectives of this study were to examine success rates, predictors of success and complications for our patients who had undergone tracheoesophageal punctures (TEPs) and voice prosthesis placement after laryngectomy. To our knowledge, this is the first analysis of the use and complications of TEPs in Singapore. PATIENTS AND METHODS: Thirty-eight patients seen in our centre were analysed in this retrospective case review. Factors analysed included voice quality with age, timing of insertion, type of closure, use of radiotherapy, regular maintenance and attendance at our clinic. Twenty-eight had undergone primary TEP surgery and 10 had undergone secondary TEP surgery for voice restoration after laryngectomy for cancer. Thirty-five patients had primary closure of the pharynx with 13 vertical, 19 horizontal and 3 T-shaped closures. The other 3 patients required reconstructive surgery. Thirty-five patients had radiotherapy. A numerical assessment of voice production was made of patients immediately and at 6 months post-insertion. A review of the complications was also done. RESULTS: TEPs provided a fair to good voice in 74% of our laryngectomees. Patients who attended the voice restoration clinics and who regularly cleaned their prostheses were found to have statistically better voices (P = 0.044 and P = 0.002, respectively). Patients less than 60 years old had better results, as did patients with horizontal or T-shaped closures; however, these were not statistically significant. Secondary TEPs provided fair/good voices in 90% of cases compared to 68% of primary TEPs. Voice quality during radiotherapy was diminished but recovered at 6 months postradiotherapy. The commonest complications noted included crusting (seen in 40% of cohort), candida infections and leaks which were seen in 16% of our patients. There was no mortality attributable to the use of TEPs in our study. PMID- 11056775 TI - Spectrum of abnormal mammographic findings and their predictive value for malignancy in Singaporean women from a population screening trial. AB - INTRODUCTION: The ability to categorise mammographic features according to their likelihood of malignancy would be valuable in the management of women with abnormal mammograms. The aim of our study was to correlate abnormal mammographic features in a screened population with their histology to identify those features which are predictive of malignancy. The study also examined the spectrum of mammographic features in an Asian population. MATERIALS AND METHOD: This prospective study involved 28,231 women who were randomly selected from a population registry and underwent two-view screening mammography without physical examination. Women with suspicious lesions were recalled for further mammographic views or to a joint assessment clinic prior to biopsy. Mammographic abnormalities and their corresponding histology were assessed. RESULTS: The spectrum of mammographic abnormalities was similar to that in Caucasian populations. The positive predictive value for malignancy was 44.1% of all biopsied cases. Mammographic features could be broadly classified into low-, moderate- and high risk categories for malignancy. Those features which correspond to high malignancy rates (9.8% to 16.0%) include multiple abnormalities or parenchymal lesions with microcalcifications. The presence of microcalcifications was a good predictor of ductal carcinoma-in-situ (DCIS): 46% of lesions in which the microcalcifications were the sole abnormality were DCIS only. Further, 71% of cancers with any microcalcification on the mammogram had a focus of DCIS on histology. CONCLUSION: Mammographic abnormalities can be segregated into three risk groups for malignancy, and this in turn can improve the selection criteria for breast biopsy, hence reducing unnecessary intervention. Furthermore, the presence of microcalcifications denotes the presence of DCIS, and would be an important determinant of the extent of surgical excision. PMID- 11056776 TI - The correction of oriental lower lid involutional entropion using the combined procedure. AB - INTRODUCTION: Involutional entropion is a common lid malposition problem in the Chinese geriatric population. The major contributing factors of involutional entropion include disinsertion of the lower lid retractors and horizontal lid laxity. The combined procedure (lower lid retractor repair and lateral tarsal strip procedure) is a useful technique that tackles both the horizontal and vertical aetiologic components of this condition. The surgical technique of this procedure is described and the results and complications presented. MATERIALS AND METHODS: A retrospective non-randomised study on 41 lower eyelids of 38 Chinese patients. The combined procedure was performed by a single surgeon over a 2 years 11 months period. Nine patients had bilateral lid surgeries. The clinical charts and operative notes were reviewed by an observer. RESULTS: The mean age of the patients was 74.7 years (range 51 to 92 years). There were 14 males (36.8%) and 24 females (63.2%). The cases were followed up postoperatively for a mean duration of 13 months (range 3 to 48 months). Twelve lids had early postoperative overcorrection but the majority (8 lids) resolved spontaneously with good lid globe apposition. Of the remaining 4 eyelids, 3 lids had persistent mild asymptomatic ectropion that did not require surgical treatment and only one required re-operation. CONCLUSION: The combined procedure is an effective means of repair for oriental lower lid entropion with low complication rates. PMID- 11056777 TI - Mental health literacy in Singapore: a comparative survey of psychiatrists and primary health professionals. AB - OBJECTIVES: To assess the beliefs amongst health professionals in Singapore about management of 3 major mental disorders, comparing psychiatrists and a sample of primary care physicians, and so identify target areas for the education of primary health professionals. MATERIALS AND METHOD: A questionnaire earlier distributed to psychiatrists at Woodbridge Hospital was posted to both Singapore general practitioners and polyclinic doctors. The questionnaire assessed the capacity of respondents to identify vignettes of depression, schizophrenia or mania, and then assessed respondents' views about the likely helpfulness of a number of interventions. RESULTS: The psychiatrists and primary health professionals differed little in terms of diagnostic accuracy for depression and schizophrenia; however, only half the general practitioners and three-quarters of the polyclinic doctors correctly diagnosed mania, which was consistently diagnosed by the psychiatrists. A number of distinct differences were identified between the groups concerning the likely helpfulness and disorder specificity of various psychotropic drugs. The primary health physicians were more likely to favour non-specific management approaches, whilst the psychiatrists generally supported a focused biological approach to treatment, especially for the psychotic disorders. Some of the differences in beliefs about mental health management may well be contributed by the different patients treated by each group of clinicians. CONCLUSIONS: The findings have important clinical implications in terms of diagnosing common psychiatric conditions accurately and giving us professionals' views about a range of interventions for such conditions, while also assisting review of educational programmes for identifying and managing major mental disorders. PMID- 11056778 TI - The reliability and validity of the Alzheimer's Disease Assessment Scale Cognitive Subscale (ADAS-Cog) among the elderly Chinese in Hong Kong. AB - INTRODUCTION: The Alzheimer's Disease Assessment Scale cognitive subscale (ADAS cog) was reported to be a sensitive cognitive function assessment scale for Alzheimer's Disease (AD). The English, Greek, Spanish but not Chinese versions had been validated previously. OBJECTIVES: The objectives of the present study were to investigate the reliability and validity of an adapted Chinese version of the ADAS-cog among Chinese elderly AD patients in Hong Kong. MATERIALS AND METHOD: Thirty-nine subjects were recruited during the period July to December 1998. Twenty were AD patients while 19 were non-demented normal subjects. Two raters administered the ADAS-cog scale thrice on different occasions. RESULTS: The internal consistency (Cronbach's alpha) of the ADAS-cog were 0.91, 0.88 and 0.65 for the whole group, the AD and normal (i.e. non-demented) subjects respectively. The test-retest reliability as measured by the Spearman's rho correlation coefficients were 0.96, 0.86 and 0.86 for the whole group, AD and normal subjects, respectively, (all P < 0.001). The Spearman's rho correlation coefficients for inter-rater reliability were 0.95 (P < 0.001), 0.91 (P < 0.001) and 0.65 (P = 0.003) for the whole group, AD and normal subjects, respectively. The ADAS-cog score was inversely related to the Mini-Mental Status Examination (MMSE) score (Spearman's rho = -0.91; P < 0.001). The ADAS-cog score was directly proportional to the Clinical Dementia Rating (CDR) (rho = 0.89; P < 0.001). Forward stepwise discriminant function analysis between AD and normal subjects yielded a canonical discriminant function with 3-question items (i.e. word recall test, orientation and comprehension of speech; P < 0.001). This short version had a sensitivity of 90%, specificity of 94.7% and overall accuracy of 92.3%. CONCLUSION: The Chinese version of ADAS-cog subscale is both reliable and valid among the elderly Chinese in Hong Kong. PMID- 11056779 TI - Perioperative deaths: a further comparative review of coroner's autopsies with particular reference to the occurrence of fatal iatrogenic injury. AB - INTRODUCTION: In previous triennial reviews of Coroner's perioperative autopsies conducted during the periods 1989 to 1991 and 1992 to 1994, it was observed that the necropsy incidence of such deaths rose from 2% to 2.6% (P < 0.05). Concurrently, the rate of iatrogenic deaths had nearly doubled from 15.2% to 28.8% (P < 0.02). These findings spurred a review of the subsequent triennium (1995 to 1997), in order to monitor the apparent rise in these trends and to study the frequency and occurrence of iatrogenic deaths in relation to the number of invasive procedures performed, as well as during emergency and elective procedures. MATERIALS AND METHODS: A retrospective (descriptive and comparative) study, comprising a clinico-pathological review of a series of 270 perioperative deaths (defined as deaths occurring during or after invasive therapeutic or diagnostic procedures, up to a week after discharge, and excluding cases of major trauma from suicides, homicides, as well as road and industrial accidents) reported to the Coroner, for which autopsies were conducted at the Department of Forensic Medicine from 1995 to 1997. RESULTS: The necropsy incidence of 4.4% (270/6074) represented a significant rise over the previous triennia (P < 0.01). As in previous years, there was a predominance of males (M:F = 1.65:1) and middle aged to elderly patients (range 0 to 92 years, mean 55.8 years, median 63 years), most of whom had died after a variable, but usually brief, postoperative interval [0 to 97, 4.2, 1 day(s)] and a more variable period of hospitalisation (< 1 to 289, 12.6, 7 days). A total of 408 invasive procedures were performed, amounting to an average of 1.5 per patient; 101 patients (37.4%) underwent multiple (> 1) interventions, which were initially classified as elective procedures in 27 cases. There were 66 (24.4%) iatrogenic deaths, of which 2 (0.7%) were due to anaesthetic mishaps; 18/64 iatrogenic deaths, unrelated to anaesthesia, occurred after the first postoperative day. The proportions of such deaths amongst patients subjected to multiple interventions, or initial elective procedures, were more than twice as high as amongst those undergoing single procedures, and those initially classified as emergencies (35.6% versus 16.6% and 33.3% versus 13.2%, respectively; P < 0.01). Only 51/66 (77.3%) iatrogenic deaths received Coroner's verdicts of misadventure; no verdict of criminal negligence was recorded during the period in question. CONCLUSIONS: There appears to have been a steady increase in the number of perioperative deaths reported to the Coroner over the previous triennia (1989 to 1997) for which autopsies were conducted. While this observation may not denote an increase in perioperative morality rates per se, it may be indicative of an increasingly "aggressive" or defensive approach to the clinical management of seriously ill patients, particularly over the past decade. Although the rate of iatrogenic deaths appears to have stabilised, it is too early to say whether this apparent trend will persist in the future. It is perhaps not surprising that the risk of iatrogenic injury appears to increase with the number of interventions performed; however, it is not clear why initial, supposedly elective, interventions should be associated with an apparently greater risk of iatrogenic injury than those classified as emergency procedures. The substantial divergence between the autopsy finding of an iatrogenic death and the corresponding Coroner's verdict of misadventure may be comforting to clinicians, but certainly warrants further examination. PMID- 11056780 TI - Theory and practice in continuing medical education. AB - INTRODUCTION: Continuing medical education (CME) represents the final and often most poorly understood stage of physician education. The understanding of contemporary theories of physician education and characteristics of effective CME interventions will help CME providers and physician learners to plan productive CME activities and improve learning. This article aims to provide readers with emerging evidences on effective CME, particularly in relation to theories of physician learning and their implications for CME planning. The article also summarises attributes of effective CME interventions. METHODS: The data and evidence were collected from contemporary medical education journals and published books on medical education. Two electronic databases, Medline and ERIC (Educational Research Information Clearinghouse) were searched for suitable articles. RESULTS: Physician learning is a distinct phenomenon with high inclination towards autonomy and self-directed learning. CME interventions are more likely to be fruitful if they are modelled with strong theoretical background, catered towards individual learning needs and preferences, and focused on the learning component of education. Many widely practised CME interventions fail to be effective as those are not based on the above principles. CONCLUSION: Evidence suggests that careful planning and evaluation of CME will improve the key measure of physician's performance and health care outcome. PMID- 11056781 TI - Analytical static stress analysis of first cervical vertebra (atlas). AB - INTRODUCTION: Fracture of the atlas was first described by Jefferson (1920). He theorised a bursting mechanism of fracture as the occipital condyles were driven into the atlas. Experimental studies by Hays and Alker (1988) and Panjabi et al (1991) were also conducted to explain the injury mechanisms. Injury mechanisms and fracture patterns are important in the clinical evaluation of spinal injuries. Recognition and interpretation of the fracture patterns help to determine the spinal instability and consequently the choice of treatment. Although the fracture mechanics of the atlas have received much attention, it has not been investigated using theoretical modelling. MATERIALS AND METHODS: A high definition digitiser was used to obtain the geometrical data for the finite element mesh generation. Contrary to the widely used method, such as computed tomography scan for geometric extraction, the direct digitising process of the dried specimen reliably preserves the accurate topography of up to 0.1-mm interval of the original structure. The finite element model was exercised under an axial compressive mode of pressure loading to investigate the sites of failure reported in vivo and in vitro. RESULTS: Using material properties from literature, the predicted results from the 7808-finite element model demonstrate high concentration of localised stress at the anterior and posterior arch of the atlas, which agrees well with those reported in the literature. Furthermore, our results are also in good agreement with the findings reported by Panjabi et al (1991), which show that the groove of the posterior arch is subjected to enormous bending moment under simulated hyperextension conditions. CONCLUSIONS: The close agreement of the failure location provided confidence to perform further analysis and in vitro experiments. The predicted results from finite element analysis may be potentially used to supplement experimental research in understanding the clinical biomechanics of the C1. PMID- 11056782 TI - A case report on the perinatal management of a 30-week preterm baby with congenital complete heart block. AB - INTRODUCTION: Congenital complete heart block is an uncommon condition in the newborn, but is known to occur with maternal systemic lupus erythematosus. CLINICAL PICTURE: This paper presents one such baby with complete heart block who was born premature (after a gestation of 30 weeks) and weighing 759 g. TREATMENT: Continuous isoprnaline infusion was initially used to support the baby while her other neonatal problems were treated. A Medtronics VV1 pacemaker was subsequently inserted to maintain a heart rate that would be more physiologically acceptable for the patient. OUTCOME: This baby is currently thriving well, having been followed up for one year. CONCLUSIONS: The management issues, encompassing maternal and neonatal problems, and a review of current literature on this condition are discussed. PMID- 11056783 TI - Sandhoff disease--a case report of 3 siblings and a review of potential therapies. AB - INTRODUCTION: Sandhoff disease is a GM2 gangliosidosis that may present within the first 6 months of life with developmental regression. This is the first report of a pedigree from Southeast Asia. CLINICAL PICTURE: All the affected siblings presented in the first year of life with developmental regression, spasticity, seizures and loss of vision. The diagnosis was confirmed by an enzymatic deficiency in both beta-hexosaminidase A and B. CONCLUSION: As the disorder is autosomal recessive, and no curative therapy is currently available, genetic counselling is necessary to prevent the burden of this devastating disease. We review the potential strategies of treatment for Sandhoff disease. PMID- 11056784 TI - A case report of the use of magnesium sulphate during anaesthesia in a patient who had adrenalectomy for phaeochromocytoma. AB - INTRODUCTION: Patients with phaeochromocytoma have haemodynamic instability during adrenalectomy. CLINICAL PICTURE: A case showing major swings of blood pressure during tumour handling. TREATMENT: Magnesium sulphate infusion alone failed to prevent severe hypertension. OUTCOME: The patient had to be given phentolamine and sodium nitroprusside to control the severe hypertension. CONCLUSION: The greatest value of magnesium sulphate is in controlling catecholamine release at induction and intubation, and in association with other agents in controlling arrhythmias and hypertension during tumour handling. PMID- 11056785 TI - Hard metal lung disease--the first case in Singapore. AB - INTRODUCTION: We report the first case of hard metal lung disease in Singapore and the occupational investigative work and control measures that were undertaken. CLINICAL PICTURE: A 38-year-old machinist in the tool manufacturing industry presented with exertional dyspnoea and cough. Chest X-ray revealed bilateral reticulonodular infiltrates with honeycombing. High resolution computed tomography scan of the thorax confirmed the presence of interstitial fibrosis. Open biopsy of the lung showed features of pneumoconiosis. Particle induced X-ray emission (PIXE) analysis, a relatively new elemental analysis technique, performed on the lung biopsy specimen confirmed the presence of tungsten and titanium; and he was diagnosed to have hard metal lung disease. Microbiologic, serologic and histologic investigations excluded an infective cause. Serial pulmonary function tests on follow-up showed no progression. He presented with haemoptysis 10 months later and was diagnosed to have tuberculosis on the basis of positive sputum and bronchoalveolar lavage cultures for Mycobacterium tuberculosis complex. TREATMENT: Preventive measures and permanent transfer to non-cobalt work were instituted. OUTCOME: The interstitial fibrosis appears to have stabilised. CONCLUSION: The diagnosis of hard metal lung disease must be considered in a worker exposed to cobalt presenting with interstitial fibrosis. PMID- 11056786 TI - Treatment of chronic urticaria with thyroxine in an euthyroid patient with thyroglobulin and microsomal antibodies. AB - INTRODUCTION: The association of chronic urticaria and thyroid autoimmunity is not well recognised and the potential use of thyroxine in the treatment of chronic urticaria in patients with thyroid autoimmunity is even less well known. CLINICAL PICTURE: We report a case of chronic urticaria in an euthyroid patient with evidence of significantly elevated levels of thyroglobulin and microsomal antibodies. TREATMENT AND OUTCOME: Treatment with thyroxine has brought about clinical remission of the chronic urticaria but no change in the thyroid antibody levels could be demonstrated. CONCLUSION: Patients with chronic urticaria should be screened for evidence of thyroid autoimmunity. A closely monitored trial of thyroxine therapy for those who have thyroid autoimmunity can be rewarding. PMID- 11056787 TI - Pseudomembranous tracheobronchitis caused by Aspergillus in a patient after peripheral blood stem cell transplantation. AB - INTRODUCTION: We report a case of pseudomembranous tracheobronchitis caused by Aspergillus fumigatus 2 years after matched unrelated stem cell transplant. CLINICAL PICTURE: The patient presented with dyspnoea and obstructive airway disease coinciding with the onset of chronic graft-versus-host disease (GVHD). Following treatment with higher immunosuppressive therapy for presumptive diagnosis of bronchiolitis obliterans, he subsequently developed recurrent spontaneous pneumomediastinum and progressive respiratory failure. TREATMENT AND OUTCOME: Tracheobronchial biopsy and culture of bronchoalveolar lavage (BAL) fluid revealed Aspergillus tracheobronchitis. Despite mechanical ventilation and antifungal therapy, he succumbed to progressive respiratory failure. CONCLUSIONS AND CLINICAL IMPLICATIONS: Aspergillus tracheobronchitis should be suspected in heavily immunosuppressed stem cell transplant recipients presenting with recurrent pneumomediastinum and progressive respiratory failure. PMID- 11056788 TI - Case reports: the use of intermaxillary screws to achieve intermaxillary fixation in the treatment of mandibular fractures. AB - INTRODUCTION: Treatment of mandibular fractures commonly involves the use of arch bars in temporary maxillo-mandibular fixation (also called intermaxillary fixation) to aid bone plating, for the postoperative application of light elastic traction to correct minor occlusal discrepancies and intermaxillary fixation for post reduction immobilisation. The purpose of this paper is to describe a quick and simple alternative for intermaxillary fixation. CLINICAL PICTURE: Three cases of mandibular fractures are presented to demonstrate the use of intermaxillary screws instead of conventional arch-bar wiring. TREATMENT: Two cases were treated with closed reduction methods while the other case was treated with open reduction. OUTCOME: Subsequent fracture healing was not compromised using this technique. CONCLUSIONS: With careful case selection, successful treatment outcome may be achieved using this time-saving and technically simple procedure. Some of the advantages and disadvantages of the intermaxillary screws are discussed. PMID- 11056789 TI - Re: One-year review of pityriasis rosea at the National Skin Centre, Singapore. PMID- 11056790 TI - Rapid-cycle PCR in temporarily compartmentalized capillaries: two-round PCR in a single capillary prevents product carry-over. PMID- 11056791 TI - Pre-PCR gel-loading buffer that increases specificity. PMID- 11056792 TI - Modified method to detect PCR products by 5' nuclease activity and an asymmetric fluorogenic probe set. PMID- 11056793 TI - Method for avoiding PCR-inhibiting contaminants when eluting DNA from polyacrylamide gels. PMID- 11056794 TI - PCR-based setup for high-throughput cDNA library sequencing on the ABI 3700 automated DNA sequencer. PMID- 11056795 TI - Increased sensitivity of one-tube, quantitative RT-PCR. PMID- 11056796 TI - Estimation of relative mRNA content by filter hybridization to a polyuridylic probe. PMID- 11056797 TI - Reliable and reproducible method to extract high-quality RNA from plant tissues rich in secondary metabolites. PMID- 11056798 TI - Secondary transient transfection of stably transfected endothelial cells. PMID- 11056799 TI - GFP and beta-galactosidase transformation vectors for promoter/enhancer analysis in Drosophila. PMID- 11056800 TI - Treatment by methyl methanesulfonate induces up-regulation of cytomegalovirus immediate/early promoter. PMID- 11056801 TI - Detection of antibody display phage without clearing of bacterial culture. PMID- 11056802 TI - Improved method for the isolation and visualization of terminal protein-bound DNA fragments in actinomycetes. PMID- 11056803 TI - Unique adaptor design for AFLP fingerprinting. PMID- 11056804 TI - Online biological research product information and sales. PMID- 11056805 TI - Poly(A) cDNA-specific (PACS) RT-PCR: a quantitative method for the measurement of any poly(A)-containing mRNA not affected by contaminating genomic DNA. AB - We present a simple and efficient RT-PCR method for the detection and quantitation of any poly(A)-containing mRNA that is not affected by contaminating genomic DNA and does not rely on exhaustive DNase digestion protocols. The technique described here requires the use of an antisense primer designed to contain 6-8 bp cDNA-specific sequence and an additional 17 Ts located on the 5' end to take advantage of the poly(A) tail. A second cDNA-specific sense primer can be used that does not need to be separated by intronic DNA sequence. PMID- 11056806 TI - Morphological selection of parental Chinese hamster ovary cell clones exhibiting high-level expression of recombinant protein. AB - To facilitate the establishment of recombinant Chinese hamster ovary (rCHO) cell lines with dihydrofolate reductase (dhfr)-mediated gene amplification, a primary selection method based on morphology of parental CHO clones has been developed. Morphology of parental clones that were made by transfecting various plasmids encoding thrombopoietin (TPO) and its analogs and humanized antibodies into dhfr deficient (dhfr-) CHO cells was not uniform. Morphology of many parental clones exhibiting high-level expression of the introduced gene was similar to that of nontransfected dhfr- CHO cells. On the other hand, most parental clones with low level expression experienced noticeable morphological changes such as bipolar fibroblast-like morphology. In case of selection of parental clones with TPO expression level higher than 200 ng/mL, morphological selection improved selection efficiency by 3.5-fold compared with random selection. Furthermore, when subjected to methotrexate for gene amplification, parental clones that were selected based on morphology elevated the expression level as much as those that were selected randomly. Taken together, morphological selection of parental clones can facilitate the establishment of rCHO cell lines expressing recombinant proteins. PMID- 11056807 TI - Quantitation of phosphotyrosine signals in human prostate cell adhesion sites. AB - A simple method is described for the quantitation of phosphotyrosine signaling in human prostate cell cultures. The phosphotyrosine signals are observed by standard immunohistochemistry techniques, and the resulting digital images are analyzed using the Scion image software program. The signals within the cell adhesion sites are quantitated using the density slice and particle analysis features of the software. The immunohistochemistry results are compared with detection of phosphotyrosine signals using a standard Western blotting procedure with whole cell lysates. The resulting data is converted into graphs using the Sigma Plot Program. This method is illustrated using damage-induced signaling within cell adhesion sites after a low dose of ionizing radiation. PMID- 11056808 TI - Pitfall of an internal control plasmid: response of Renilla luciferase (pRL-TK) plasmid to dihydrotestosterone and dexamethasone. AB - The thymidine kinase promoter-Renilla luciferase reporter plasmid (pRL-TK) is commonly used as a control for transfection efficiency in the Dual-Luciferase Reporter Assay System. While investigating hormone response elements in the promoters of the androgen-dependent, epididymis-specific EP2 gene, we found that hormone treatment affected the luciferase activity of pRL-TK-transfected cells. In African Green Monkey Kidney (CV-1) cells, cotransfected transiently with a hormone-responsive promoter-firefly luciferase reporter plasmid and with pRL-TK, Renilla luciferase activity increased in response to dihydrotestosterone (DHT) and decreased in response to dexamethasone (DEX). When a thromboxane synthase promoter Renilla luciferase plasmid (pRL-TS) was used in place of pRL-TK, Renilla luciferase activity remained constant in CV-1 cells treated with DHT but decreased in CV-1 cells treated with DEX. In transfection studies, internal control plasmid expression in response to treatment must be carefully monitored to ensure proper interpretation of normalized results. PMID- 11056809 TI - Magnetic, microplate-format plasmid isolation protocol for high-yield, sequencing grade DNA. AB - We have developed a rapid, microplate-format plasmid isolation procedure to purify sequencing-grade DNA templates for high-throughput DNA sequencing operations. A modified lysozyme/boiling method is used to produce a plasmid containing supernatant that is then purified by iron bead capture. After binding, the beads are pelleted in a magnetic field, washed and the DNA eluted in water. The method yields up to 10 micrograms plasmid DNA from a 1-mL overnight culture in a deep-well microplate. The procedure is suitable for large-scale experiments, amenable to automation and does not require expensive reagents or equipment. The entire protocol can be completed in as little as 2 h, and one technician with a 96-well pipetting station can process up to 48 plates per day. This protocol is ideal for any high-throughput operation in which template quantity, quality and reproducibility are of primary importance. PMID- 11056810 TI - Method for linking a synthesized protein to its mRNA-DNA complex. AB - A nascent protein remains in a complex with its ribosome and mRNA if the stop codon is deleted from the mRNA. In the same manner, mRNA forms a stable complex with DNA if the transcription termination is blocked. In principle, if both mRNA translation and DNA transcription termination are prevented, the protein should stay in a complex with its mRNA and DNA. A method is designed to test these possibilities. Using an immobilized luciferase gene sequence, a functional luciferase protein is produced that remains associated through its mRNA with its DNA, confirming the feasibility of the proposed scheme. It has potential application for in vitro synthesis of proteins and protein micro-arrays. PMID- 11056811 TI - Fluorescent protein vector for directional selection of PCR clones. AB - Green fluorescent protein (GFP) has become a convenient and versatile tool as a reporter protein in many aspects of science. Here, we show that the enhanced yellow fluorescent protein (EYFP) variant may be used advantageously as a reporter system for directional cloning of blunt-ended PCR products. We have constructed a pUC18-derived plasmid containing a reporter gene coding EYFP cloned into the BamHI/HindIII sites. The blunt-ended PCR product is cloned into the SmaI site of that plasmid. A reverse PCR primer must be designed with extra bases on the 5' end that are required to introduce a ribosome binding site (rbs) for EYFP expression. The reporter gene coding EYFP is not expressed unless an rbs is introduced in the proper orientation at the 3' end of the cloned PCR insert. The results of this cloning procedure may be analyzed by simple visual inspection using a transilluminator. In most cases, successful directional cloning results in white fluorescent colonies. The proposed procedure is a convenient method that can reduce the time- and labor-intensive analysis of the clones obtained during blunt-ended PCR product cloning. PMID- 11056812 TI - Herpes simplex virus-enhanced cationic lipid/DNA-mediated transfection. AB - Liposome plasmid DNA complexes (lipoplexes) are often inefficient in mediating gene transfer and expression because of DNA degradation in lysosomal vesicles. Because herpes simplex virus (HSV) enters cells by fusion of the virus envelope with the plasma membranes, thereby overriding the endosomal pathway, HSV/lipoplex mixtures could be useful for improving gene transfer particularly when the mixture uses highly defective HSV particles that fail to express cytotoxic viral gene products. To evaluate this possibility, lipoplexes composed of cationic liposomes and lacZ reporter plasmids were compared for their ability to transduce cells in culture in the presence and absence of infectious HSV particles. The results showed that HSV increased the efficiency of cell transduction by approximately 4-100-fold compared with lipoplex vector alone, depending on the cell type targeted for gene delivery. The increased efficiency of transduction was virus dose dependent and required virus entry. PMID- 11056813 TI - Use of an IRES bicistronic construct to trace expression of exogenously introduced mRNA in zebrafish embryos. AB - To understand gene function in developing vertebrate embryos, co-injection of an mRNA for a reporter protein and an mRNA for a testing factor is widely used. However, because of the mosaic segregation of injected nucleic acids during early embryogenesis, whether both mRNAs are translated in the same cell remains uncertain. In the present study, we tested a new system of tracing the expression of a testing gene in zebrafish using an internal ribosomal entry site (IRES) to express two proteins from the same mRNA template, thus eliminating the problem of independent translation observed in co-injection essays. A DNA construct was made for synthesizing bicistronic mRNA for NeuroD, a neurogenic transcription factor, and the enhanced green fluorescent protein (EGFP) reporter. When the bicistronic mRNA for NeuroD and EGFP was injected into zebrafish embryos at one cell stage, all EGFP-expressing embryos showed ectopic expression of neuroD mRNA and the mRNA of its potential downstream gene, islet-1. Thus, the IRES bicistronic mRNA construct might be a more convincing means of analyzing gene function in developing zebrafish embryos. PMID- 11056814 TI - Human antimicrobial peptides: analysis and application. AB - Antimicrobial peptides are innate host defense molecules that have a direct effect on bacteria, fungi and enveloped viruses. They are found in evolutionarily diverse species ranging from prokaryotes and plants to invertebrate and vertebrate animals. Humans express several families of antimicrobial peptides in myeloid cells and on various epithelial surfaces where they are poised to defend against pathogens. Recently, antimicrobial peptides from animals and plants have served as templates for the design of new therapeutic antibiotics. This review provides an introduction to the biology of human antimicrobial peptides, followed by a more detailed discussion of their isolation from tissues and biological fluids, their purification by gel electrophoresis and chromatography and assays of their antimicrobial activities. PMID- 11056815 TI - Transgenic plants and biosafety: science, misconceptions and public perceptions. AB - One usually thinks of plant biology as a non-controversial topic, but the concerns raised over the biosafety of genetically modified (GM) plants have reached disproportionate levels relative to the actual risks. While the technology of changing the genome of plants has been gradually refined and increasingly implemented, the commercialization of GM crops has exploded. Today's commercialized transgenic plants have been produced using Agrobacterium tumefaciens-mediated transformation or gene gun-mediated transformation. Recently, incremental improvements of biotechnologies, such as the use of green fluorescent protein (GFP) as a selectable marker, have been developed. Non transformation genetic modification technologies such as chimeraplasty will be increasingly used to more precisely modify germplasm. In spite of the increasing knowledge about genetic modification of plants, concerns over ecological and food biosafety have escalated beyond scientific rationality. While several risks associated with GM crops and foods have been identified, the popular press, spurred by colorful protest groups, has left the general public with a sense of imminent danger. Reviewed here are the risks that are currently under research. Ecological biosafety research has identified potential risks associated with certain crop/transgene combinations, such as intra- and interspecific transgene flow, persistence and the consequences of transgenes in unintended hosts. Resistance management strategies for insect resistance transgenes and non-target effects of these genes have also been studied. Food biosafety research has focused on transgenic product toxicity and allergenicity. However, an estimated 3.5 x 10(12) transgenic plants have been grown in the U.S. in the past 12 years, with over two trillion being grown in 1999 and 2000 alone. These large numbers and the absence of any negative reports of compromised biosafety indicate that genetic modification by biotechnology poses no immediate or significant risks and that resulting food products from GM crops are as safe as foods from conventional varieties. We are increasingly convinced that scientists have a duty to conduct objective research and to effectively communicate the results--especially those pertaining to the relative risks and potential benefits--to scientists first and then to the public. All stakeholders in the technology need more effective dialogues to better understand risks and benefits of adopting or not adopting agricultural biotechnologies. PMID- 11056816 TI - PCR amplification on a microarray of gel-immobilized oligonucleotides: detection of bacterial toxin- and drug-resistant genes and their mutations. AB - PCR amplification on a microarray of gel-immobilized primers (microchip) has been developed. One of a pair of PCR primers was immobilized inside a separate microchip polyacrylamide porous gel pad of 0.1 x 0.1 x 0.02 (or 0.04) micron in size and 0.2 (or 0.4) nL in volume. The amplification was carried out simultaneously both in solution covering the microchip array and inside gel pads. Each gel pad contained the immobilized forward primers, while the fluorescently labeled reverse primers, as well as all components of the amplification reaction, diffused into the gel pads from the solution. To increase the amplification efficiency, the forward primers were also added into the solution. The kinetics of amplification was measured in real time in parallel for all gel pads with a fluorescent microscope equipped with a charge-coupled device (CCD) camera. The accuracy of the amplification was assessed by using the melting curves obtained for the duplexes formed by the labeled amplification product and the gel immobilized primers during the amplification process; alternatively, the duplexes were produced by hybridization of the extended immobilized primers with labeled oligonucleotide probes. The on-chip amplification was applied to detect the anthrax toxin genes and the plasmid-borne beta-lactamase gene responsible for bacterial ampicillin resistance. The allele-specific type of PCR amplification was used to identify the Shiga toxin gene and discriminate it from the Shiga-like one. The genomic mutations responsible for rifampicin resistance of the Mycobacterium tuberculosis strains were detected by the same type of PCR amplification of the rpoB gene fragment isolated from sputum of tuberculosis patients. The on-chip PCR amplification has been shown to be a rapid, inexpensive and powerful tool to test genes responsible for bacterial toxin production and drug resistance, as well as to reveal point nucleotide mutations. PMID- 11056817 TI - Modified AFLP technique for rapid genetic characterization in plants. AB - The standard amplified fragment-length polymorphism (AFLP) technique was modified to develop a convenient and reliable technique for rapid genetic characterization of plants. Modifications included (i) using one restriction enzyme, one adapter molecule and primer, (ii) incorporating formamide to generate more intense and uniform bands and (iii) using agarose gel electrophoresis. Sea oats (Uniola paniculata L.), pickerel-weed (Pontederia cordata L.), Bermudagrass (Cynodon dactylon L.) and Penstemon heterophyllus Lindl. were used to determine the ability to generate adequate resolution power with both self- and cross pollinated plant species including cultivars, ecotypes and individuals within populations. Reproducibility of bands was higher in all the AFLP experiments compared to random amplified polymorphic DNA (RAPD). Formamide with or without bovine serum albumin improved band intensities compared to dimethyl sulfoxide and the standard reaction mixture with no organic solvents. Comparison between RAPD and modified AFLP using sea-oats population samples proved that modified AFLP exhibits (i) a low number of faint bands with increased specificity of amplified bands, (ii) a significantly higher number of polymorphic loci per primer, (iii) less primer screening time, (iv) easy scoring associated with fewer faint bands and (v) greatly enhanced reproducibility. The technique described here can be applied with a high degree of accuracy for plant genetic characterization. PMID- 11056818 TI - Microsatellite genotyping of post-PCR fluorescently labeled markers. AB - We show that a post-PCR multicolor fluorescence-labeling technique is applicable to multiplex microsatellite genotyping. Forty-three dinucleotide microsatellite markers, which are located on 11q13-23, a candidate region for dominant familial exudative vitreoretinopathy (FEVR), were used to evaluate the quality of the marker profile produced by this technique. Thirty-eight people from six families with this disease were subjects for genotyping. The samples revealed clearly separated fragment peaks with signal intensities sufficient for analysis. All genotypes were consistent with Mendelian inheritance within each family. This method is cost effective because it does not use expensive fluorescently labeled primers. It also has the advantage of avoiding ambiguity in the analysis, which may arise from the addition of non-template nucleotides by Taq DNA polymerase. PMID- 11056819 TI - Automated image analysis of live/dead staining of the fungus Aureobasidium pullulans on microscope slides and leaf surfaces. AB - An image analysis program and protocol for the identification and enumeration of live versus dead cells of the yeast-like fungus Aureobasidium pullulans was developed for both populations on microscope slides and leaf surfaces. Live cells took up CellTracker Blue, while nonviable cells stained with DEAD Red. Image analysis macro programs running under Optimas software were used to acquire images and to differentiate and enumerate viable from nonviable cells. The software was capable of discriminating green as a third parameter for identification and quantification of green fluorescent protein-expressing cells in a wild-type population. PMID- 11056820 TI - Improved recombinant retroviral titers utilizing trichostatin A. AB - Recombinant retroviruses are a common vehicle to deliver an exogenous gene to a target cell, which makes them a useful tool in the field of gene therapy. A major drawback to using recombinant retroviruses is the low titer achieved, resulting in a limited number of target cells infected and subsequently poor expression of a transgene. In this study, we created an ecotropic producer cell line that contained recombinant mouse nerve growth factor (NGF) and the reporter gene LacZ. This cell line, termed psi 2/LIG/NGF, was treated with the drug trichostatin A, an inhibitor of histone deacetylase. At a concentration of 3 microM trichostatin A, the retroviral titer of this producer cell line was increased 34-fold relative to untreated control cells and 3.4-fold compared to the commonly used hyperacetylating compound sodium butyrate. Producer cells treated with trichostatin A also increased the number of primary rat marrow stromal cells infected 5.8-fold compared to untreated producer cells. These results offer a simple and effective solution for converting low titer producer cell clones to higher titer clones, which can be easily tested and applied. PMID- 11056821 TI - Use of high specific activity StarFire oligonucleotide probes to visualize low abundance pre-mRNA splicing intermediates in S. pombe. AB - An oligonucleotide labeling system was developed that can produce radiolabeled hybridization probes with tenfold or more higher specific activity than is obtained by traditional 5'-end-labeling with polynucleotide kinase. Yet the system is as rapid and simple as kinase labeling. The reaction uses the Klenow fragment of E. coli DNA polymerase to add alpha-32P-dA residues to the 3'-end of an oligonucleotide in a primer-extension reaction. Unlike other methods of radioactive tailing (e.g., terminal transferase), a single species is produced of both known length and known specific activity. The reaction is efficient, and over 90% of probe molecules are routinely labeled. Using this method of labeling, an oligonucleotide was shown to be tenfold more sensitive in detecting target DNA sequences in a dot blot hybridization assay, compared to the same oligonucleotide labeled using polynucleotide kinase. Northern blots of Schizosaccharomyces pombe RNA were probed with an oligonucleotide specific for intron 1 of the tf2d gene, a TATA-box binding transcription factor. Kinase-labeled tf2d probe detected only unspliced RNA, while the same oligonucleotide labeled using the new method detected both unspliced tf2d RNA and rare pre-mRNA splicing intermediates. PMID- 11056822 TI - Aboriginal mental health--moving forward. PMID- 11056823 TI - The mental health of Aboriginal peoples: transformations of identity and community. AB - This paper reviews some recent research on the mental health of the First Nations, Inuit, and Metis of Canada. We summarize evidence for the social origins of mental health problems and illustrate the ongoing responses of individuals and communities to the legacy of colonization. Cultural discontinuity and oppression have been linked to high rates of depression, alcoholism, suicide, and violence in many communities, with the greatest impact on youth. Despite these challenges, many communities have done well, and research is needed to identify the factors that promote wellness. Cultural psychiatry can contribute to rethinking mental health services and health promotion for indigenous populations and communities. PMID- 11056824 TI - Mental health services for American Indians and Alaska Natives: need, use, and barriers to effective care. AB - This special review summarizes and illustrates the state of our knowledge regarding the mental health needs of American Indians and Alaska Natives. These needs are considerable and pervasive. The discussion begins by reflecting on the limits of psychiatric nomenclature and conceptual frameworks for revealing Native constructions of mental health and mental illness. The experience and manifestation of psychopathology can be both different and the same across cultures, hinging upon the extent to which such basic assumptions as the relationship of mind to body--and spirit in the case of Native people--or the primacy of the individual or social collective are shared. Having set the stage, this paper moves to recent empirical evidence regarding the mental health needs of American Indians and Alaska Natives: we review that evidence and consider it within the broader context of available services. The report closes with a brief overview of the most pressing issues and forces for change afoot in Indian country in the US. Most have to do with the structure and financing of care as tribes and other Native community-based organizations seek to balance self determination and resource management to arrive at effective, fiscally responsible, culturally informed prevention, treatment, and aftercare options for their members. These changes may herald similar trends among First Nations people to the immediate north. PMID- 11056825 TI - The Clarke Institute experience with completed suicide: 1966 to 1997. AB - OBJECTIVES: To describe the caseload of completed suicides at a single psychiatric facility and to review the perceived deficiencies in the care of those patients. METHOD: Demographic and diagnostic data, clinical circumstances, and the deficiencies in care and documentation or both were extracted from medical records and post-suicide audit reports. RESULTS: There were 276 completed suicides over the period reviewed, yielding suicide rates of 206 per 100,000 registered patients and 123.5 per 100,000 inpatient discharges. The male to female ratio was 2:1, and patients with schizophrenia or depression accounted for 63.7% of the caseload. Only 18% of inpatients were involuntary, and only 10% were under individual observation at the time of suicide. Individual psychiatrists had up to 15 suicides in their caseloads. Deficiencies and recommendations pursuant to case audits are summarized. CONCLUSION: This is the first report of the entire cumulative experience with completed suicide, including audited deficiencies in the care and documentation of that caseload, at a single Canadian psychiatric facility. PMID- 11056826 TI - Controlling the environment to prevent suicide: international perspectives. AB - BACKGROUND: Suicide and suicidal behaviour are multifaceted events requiring complex solutions. Controlling the environment is a neglected solution, despite strong support for this approach from the World Health Organization (WHO). METHOD: To discuss this approach from a global view, this review is written by authors from various cultures: American, Australian, Canadian, Chinese, Cuban, Dutch, Indian, Irish, Japanese, Lithuanian, Native North American, Russian, and South African. RESULTS: We examine gun control to illustrate the environmental control approach; however, the worldwide diversity of suicide methods calls for diverse responses. Further, controlling the environment encompasses more than restricting the means of suicide, which we illustrate with examples of toned-down media reports and restricted medicine availability. CONCLUSIONS: Controlling the environment may be a viable strategy for preventing suicide, although research shows that few clinicians implement such approaches. PMID- 11056827 TI - Who uses bibliotherapy and why? A survey from an underserviced area. AB - OBJECTIVE: To assess which mental health therapists use bibliotherapy, their reasons for doing so, and rationale for recommending specific titles. To review the book selected most often in several categories, using prepublished criteria for reviewers of self-help books. METHOD: We sent a survey to all therapists in a Northern Ontario community requesting information on therapist demographics, the respondent's practice, the use of bibliotherapy, and details of the book most often prescribed in various categories. RESULTS: Of 112 surveys, 62 were returned, for a response rate of 55%. Sixty-eight percent of respondents indicated that they used bibliotherapy. The most common reason for recommending books was to encourage self-help. There was a significant relation between greater counselling experience and increased use of bibliotherapy. Three of the 5 books reviewed were based on empirical theory; only 1 met all the guidelines. CONCLUSION: Most therapists recommend books to their clients, but there is little empirical evidence of efficacy. Counsellors should review the books recommended and discuss them with the client. Client opinion should be solicited and effectiveness measured. PMID- 11056828 TI - Estimated prevalence of the seasonal subtype of major depression in a Canadian community sample. AB - OBJECTIVE: To examine estimates of lifetime prevalence of seasonal affective disorder (SAD) in Toronto, Ontario. METHOD: Random telephone numbers were generated for the city of Toronto, and 781 respondents completed a telephone interview. Trained nonphysician interviewers conducted all interviews, which involved structured questions for diagnosing major depression. Patterns of symptom change across seasons were evaluated to establish a diagnosis of SAD according to DSM-III-R criteria. RESULTS: Correcting for sex and age, the prevalence of SAD defined by DSM-III-R criteria was 2.9% (95% CI, 1.7% to 4.0%), and the overall lifetime prevalence of major depression in the sample was 26.4% (95% CI, 23.3% to 29.4%). Some subjects were contacted for a follow-up interview conducted in person; the positive predictive value for the diagnosis of major depression for the telephone interview was 100%, and the negative predictive value was 93%. CONCLUSIONS: The seasonal subtype of depression represents 11% of all subjects with major depression, suggesting that SAD is a significant public health concern. The telephone interview demonstrated adequate reliability, indicating that it is appropriate for epidemiological surveys of this nature. PMID- 11056829 TI - Training residents for community psychiatric practice: the resident perspective. AB - OBJECTIVE: To capture the views and experiences of psychiatry residents in 5 Canadian training programs with respect to community psychiatric training. METHOD: We sent a questionnaire regarding respondent demography, career interests, and community psychiatry training experiences to psychiatry residents at 5 training programs in Ontario. We undertook descriptive analyses and frequency comparisons on the returned data. RESULTS: The overall response rate was 48%, with considerable variation between programs. With respect to career planning, respondents indicated highest interest in urban hospital-based practice. An interest in additional community psychiatry training was, however, expressed, particularly by junior residents. Residents' suggestions for improving community psychiatry training included better promotion of training opportunities, improved quality of supervision in community settings, and more didactic teaching. CONCLUSION: Psychiatry residents are obtaining training experiences in community psychiatry, but objectives and guidelines are quite variable, as is reflected in their understanding of the definition of "community psychiatry." Residents' career paths are still focused on urban hospital-based practice or solo private practice, which likely reflects their prevalent training experiences. There is, however, interest in community psychiatry training. Junior residents may be more open to this type of practice, and curricula should allow more exposure to community psychiatry at this stage of training. PMID- 11056830 TI - Aphorisms in psychotherapy. PMID- 11056831 TI - Sudden remission of obsessive-compulsive disorder by involuntary, massive exposure. PMID- 11056832 TI - Olanzapine for nightmares and sleep disturbance in posttraumatic stress disorder (PTSD) PMID- 11056833 TI - New-onset diabetes mellitus associated with quetiapine. PMID- 11056834 TI - Reboxetine and treatment-refractory elderly patients with depression. PMID- 11056835 TI - Male genital self-mutilation in patients with schizophrenia. PMID- 11056836 TI - Patient-centred versus client-centred mental health care. PMID- 11056837 TI - Sources of abnormal EEG activity in brain infarctions. AB - EEGs from 16 patients with stroke in three different stages of evolution were recorded. EEG sources were calculated every 0.39 Hz by frequency domain VARETA. The main source was within the delta band in 2 out of 4 chronic patients, and in 67% of the patients in the acute or subacute stages when edema (cytotoxic or vasogenic) was present. Moreover, all patients showed abnormal activity in the theta band. Sources of abnormal activity in cortical or corticosubcortical infarcts were located in the cortex, surrounding the lesion. At the site of the infarct, a decrease of EEG power was observed. Sources of abnormal theta power coincided with edema and/or ischemic penumbra. PMID- 11056838 TI - Effect of normal aging upon interhemispheric EEG coherence: analysis during rest and photic stimulation. AB - The present study was conducted to examine and compare interhemispheric EEG coherence at rest and during photic stimulation (PS; 5, 10 and 15 Hz) in 30 younger subjects aged 22.1 +/- 2.2 (mean +/- S.D.) and 25 elderly subjects aged 56.8 +/- 4.9. The elderly subjects had significantly lower coherence in the resting EEG for the delta, theta, alpha-3, beta-1 and beta-2 frequency bands. In contrast, EEG analysis during PS showed that elderly subjects had significantly higher coherence in the frequency corresponding to PS at 15 Hz. In addition, when we examined the changes in coherence from the resting state to the stimulus condition (i.e., coherence reactivity), elderly subjects had significantly higher coherence reactivity to PS at 15 Hz. These findings suggest a low interhemispheric functional connectivity in elderly subjects under non stimulus conditions and a high connectivity during photic stimulus. PMID- 11056839 TI - Transitory cognitive impairment in epileptic children during a CPT task. AB - EEGs and behavioral responses were studied in two sex matched groups of 58 epileptic and 20 healthy children between 8 and 12 years of age, during the execution of a go-no go CPT (X; A-X) task to determine transitory cognitive impairment (TCI) incidence. Paroxysmal discharges were found on 87.9% and 5% of the EEGs in the epileptic and control groups respectively, with no differences related to sex. The predominant EEG findings with respect to paroxysmal discharges were the association of two or more types of paroxysms with frequency higher than 5/minute, an average duration less than 0.5 second and topographical distribution over temporal-parietal-occipital areas without significant interhemispheric differences. TCI was detected in 36.2% of epileptic children. The epileptic group showed significantly higher numbers of behavioral errors and longer reaction times (RTs) in relation to the control group. Analyzing RTs on the two blocks of the task, linear discriminant analysis showed an acceptable classification of TCI incidence between groups. PMID- 11056840 TI - Wavelet analysis of transient biomedical signals and its application to detection of epileptiform activity in the EEG. AB - Wavelet based signal analysis provides a powerful new means for the analysis of nonstationary signals such as the human EEG. The properties of the discrete wavelet transform are reviewed in illustrated application examples. The continuous wavelet transform is shown to provide better detection and representation of isolated transients. An approach to extract features of edges and transients from the continuous wavelet transform is outlined. Matching pursuit is presented as a more general transform method that covers both transients and oscillation spindles. A statistical model for the continuous wavelet transform of background EEG is found. A spike detection system based on this background model is presented. The performance of this detection system has been assessed in a preliminary clinical study of 11 EEG recordings containing epileptiform activity and shown to have a sensitivity of 84% and a selectivity of 12%. The spatial context of epileptiform activity will be incorporated to improve system performance. PMID- 11056841 TI - Considerations of nonconvulsive status epilepticus. AB - The original concepts of absence status (AS) and complex partial status (CPS) are critically reviewed. This review has been prompted by a modern concept of nonconvulsive status epilepticus (NCSE), portrayed as a rather common condition occurring chiefly in the critically ill elderly with high morbidity and mortality. This new view is a striking departure from the original concepts of AS and CPS as rare protracted epileptic events occurring usually in temporarily confused but otherwise satisfactorily healthy and ambulatory patients. This new trend appears to have been caused by a misinterpretation of EEG findings: prominent generalized spike activity is in reality the expression of a very severe encephalopathy rather than of NCSE, most often caused by an anoxic episode. The role of EEG is emphasized but a valuable interpretation depends on an expert integration of EEG and clinical data. A brief discussion of epileptic twilight states further stresses the difficult differential diagnosis. PMID- 11056842 TI - EEG findings in Konzo: a spastic para/tetraparesis of acute onset. AB - EEGs were recorded on 21 konzo subjects (median age 17 years) and 13 of their close healthy relatives (median age 41 years). Konzo subjects were clinically selected and classified according to the WHO criteria. Standard waking EEG recordings were performed according to the International 10-20 System. Slowing of the background activity with theta activity was the most common abnormality (57%). The more clinically severely affected the konzo subject, the more often generalized EEG abnormalities were seen. None of these abnormalities were considered specific for konzo nor related to the duration of the disorder; however, the findings indicate involvement of the cerebral cortex. PMID- 11056843 TI - Correlation between clinical stages and EEG findings of subacute sclerosing panencephalitis. AB - In this retrospective study 67 patients with SSPE seen between the years 1980 and 1998 were reviewed. Using the criteria of SSPE diagnosis (clinical signs, characteristic EEG patterns, high titres of measles antibodies in the serum and CSF), the patients were divided into two groups. Group A fulfilled all criteria, however, due to the inability of measuring measles antibody before 1987, it was not possible to observe the third criterion in Group B. Among 67 patients, groups A and B consisted of 51 boys and 16 girls ranging in age between 1 to 23 years, mean age 13.1. The male/female ratio was 3.1. The periodic EEG complexes (PCs) were usually bilateral, synchronous and symmetrical. PC amplitude asymmetry was seen in 12 patients and 2 patients had no PC synchronization between the hemispheres. Six patients had more than one form of PC. Delta activity in anterior hemispheres after PC was seen in 40 patients, mostly in stage 2A. Thirty two patients had focal epileptiform abnormalities in multiple locations at every stage but most frequently in frontal, central and temporal regions. One patient had PC over both hemispheres and periodic lateralized epileptiform discharges (PLEDs) over the right hemisphere. The EEG findings described and observed in our study do not seem to be specific to SSPE but these findings were not atypical or unusual. PMID- 11056844 TI - The role of tiagabine in the treatment of intractable epilepsy of childhood with multifocal independent spikes: a case report. AB - Multifocal independent spike syndrome (MISS) is an identifiable electroclinical syndrome, which combines intractable motor seizures, mental retardation and multifocal independent spike discharges. Similarities to the Lennox Gastaut syndrome cause frequent misdiagnosis; however, MISS is a distinct electroclinical syndrome in children with a better prognosis and a different EEG pattern. We report an 8-year-old boy with MISS, whose intractable seizures were completely controlled for the first time with tiagabine as add-on therapy. Tiagabine should be studied further in the treatment of intractable multiple seizures of childhood. PMID- 11056845 TI - Heritability and the comorbidity of attention deficit hyperactivity disorder with behavioral disorders and executive function deficits: a preliminary investigation. AB - The heritability and comorbidity of attention deficit hyperactivity disorder (ADHD) with conduct disorder (CD), oppositional defiant disorder (ODD), and executive function (EF) deficits were examined in 224 child twins (140 monozygotic and 84 dizygotic). The Coolidge Personality and Neuropsychological Inventory for Children (Coolidge, 1998), a standardized, 200-item, Diagnostic and Statistical Manual of Mental Disorders (4th ed.; American Psychiatric Association, 1994) aligned, parent-as-respondent inventory, assessed psychopathology. Structural equation model fitting revealed that the individual scale heritabilities were substantial: .82 for ADHD, .74 for CD, .61 for ODD, and .77 for EF deficits. The results of the multivariate twin analyses suggest that ADHD shares most of its genetic liability with CD, ODD, and EF deficits. Thus, the findings argue for a common biological risk underlying these commonly comorbid externalizing behavior problems and cognitive deficits. The residual genetic variance provides preliminary support for additional genetic influences underlying CD, ODD, and EF that are independent of ADHD. PMID- 11056846 TI - School-age consequences of birth weight less than 750 g: a review and update. AB - Recent advances in perinatal care have led to the survival of increasing numbers of children born at the lower limits of viability. Children with very low birth weight (LBW; less than 1,500 g, 3 lb 5 oz) have been studied extensively. Findings document poorer outcomes relative to normal birth weight term-born controls in neurologic and health status, cognitive-neuropsychological skills, school performance, academic achievement, and behavior. This report reviews current knowledge regarding LBW children, with special emphasis on outcomes for children with birth weight less than 750 g (1 lb 10 oz). Results from an ongoing longitudinal study suggest a gradient of sequelae, with poorer outcomes in less than 750 g birth weight children compared to both 750 g to 1,499 g birth weight children and term-born controls. Children with less than 750 g birth weight fail to catch up with their peers over time and may even be at risk for age-related increases in sequelae. Outcomes are highly variable but related to neonatal medical complications of prematurity and social risk factors. Further research is needed to understand the etiology and neuropathological basis of sequelae, the long-term developmental implications of LBW, and treatment needs. PMID- 11056847 TI - Longitudinal analysis of the effects of the aging process on neuropsychological test performance in the healthy young-old and oldest-old. AB - A sample of 33 young-old (ages 65 to 74) and 20 oldest-old (ages 84 to 93) healthy elderly without dementia were assessed with neuropsychological tests annually over a 4-year period to examine longitudinal changes in cognitive functioning. Significant age-group differences existed at baseline in participants' performances on tests of immediate memory and visuospatial skills. There were no age-group differences in the rate of change over the 4-year interval on any neuropsychological tests. Within each age-group, the amount of change over time was minimal for most tests though some practice effects were apparent, and on some tests mild decline was observed. Results suggest that healthy old adults, including the oldest-old, do not experience measurable declines in cognitive functioning over a 4-year period. PMID- 11056848 TI - Etiology of neuroanatomical correlates of reading disability. AB - The heritable nature of reading disability has been well documented (DeFries & Alarcon, 1996), and possible abnormalities of brain structures have been associated with the disorder (Filipek, 1995). However, the etiology of individual differences in morphological brain measures has not been examined extensively. The purpose of this study was to apply behavioral genetic methods to assess the etiology of individual differences in neuroanatomical structures. Measures of reading performance, cognitive ability, and magnetic resonance imaging scans were obtained from 25 monozygotic (MZ) and 23 same-sex dizygotic (DZ) twin pairs with reading disability, and 9 MZ and 9 DZ control twin pairs participating in the Colorado Learning Disabilities Research Center. Results obtained from multiple regression analyses (DeFries & Fulker, 1985, 1988) of these twin data indicated that individual differences in the size of most cortical and subcortical structures were largely due to heritable influences. Moreover, estimates of heritability did not change appreciably after controlling for IQ and total brain size. PMID- 11056849 TI - Attention following pediatric head injury: a developmental perspective. AB - This study investigated the relations between age at injury (AAI) and attentional functioning, and intellectual and academic achievement following pediatric head injury. The theoretical framework of attention proposed by Mirsky, Anthony, Duncan, Ahern, and Kellam (1991) provided the basis for this analysis, and the profile of attention found in uninjured children was evaluated for this sample. Thirty-three moderately head-injured individuals were recruited, with AAI ranging from 1 to 12 years. First, the development of attentional skills in this closed head injury (CHI) sample was found to be comparable to that of Mirsky et al.'s model. Second, AAI did not predict outcome and appeared not to be associated with the finding of mildly delayed acquisition of spelling and arithmetic skills. Also, AAI did not predict the behavioral symptoms of inattention present in this sample as indicated by parental report. It may be that AAI only influences development following severe CHI in which there is permanent cerebral pathology. PMID- 11056850 TI - [Telemedicine in otorhinolaryngology. Basic principles and possible applications]. AB - Telemedicine includes all medical activities involved in diagnosis, therapeutics or social medicine undertaken by an electronic transfer medium. This technique requires the transmission of visual and acoustic information over long distances and does not require the specialist to be personally present at the requested consultation. In the last few years, the digital data transmission, e.g., ISDN (ISDN (Integrated Service Digital Network), has become available and has facilitated the use of telecommunication. Recently, the real-time transmission of acoustic and visual signals will be improved by use of asynchronous transfer mode (ATM). Advanced telecommunication applications in minimally invasive ENT surgery are experimental in most cases. We can distinguish three different telesurgical developments: surgical teleconsultation, surgical teleassistance, and surgical telemanipulation. The different applications and transmission media are explained and discussed. PMID- 11056851 TI - [Pilot study of disinfection of optical instruments in otorhinolaryngology]. AB - There is undoubtedly a need for a safe disinfection method of optical instruments used by ENT specialists. So far, a standard method that sufficiently covers the risk of infection for patients and medical staff alike while serving the need of practicability in daily routine has not been established. Therefore, in the ENT departments of the hospitals of Mannheim, Kaiserslautern, and Tubingen, studies on the disinfection of rigid optical instruments without working channel were performed between February 1999 and May 1999. Instruments were wiped either with an aldehyde or alcohol (ethanol 80%) and germs were detected by quantitative swab cultures. As discussed in the article, disinfection with ethanol proved to be at least equally efficient if not better than disinfection with aldehyde. Therefore, it fully serves the requirements of a safe method for disinfecting optical instruments without working channels. PMID- 11056852 TI - [Family study of patients with aspirin intolerance and rhinosinusitis]. AB - The high prevalence of aspirin intolerance in asthmatics and patients with nasal polyps as well as reports of familial clustering suggest a genetic disposition of this disease. Our study aimed at obtaining further evidence of hereditary factors in this disease. We included 33 unselected patients from 28 families with aspirin intolerance and rhinosinusitis in this study. Controls were recruited from individuals treated in our ENT clinic for diseases other than aspirin intolerance (n = 52). A questionnaire focused on family histories as well as reports on diseases of the upper respiratory tract or allergies. ASS intolerance was verified either by bronchial or nasal provocation tests. We found cases of aspirin intolerance among parents, siblings, and children of ASS intolerant probands. The children of probands had nasal polyps and rhinosinusitis more often than the children of controls. We propose that ASS intolerance with nasal polyps and asthma represents a complex phenotype, with genetic and environmental factors contributing to its manifestation. PMID- 11056853 TI - [Prognostic value of hemoglobin level for primary radiochemotherapy of head-neck carcinomas]. AB - The pretherapeutic hemoglobin level (Hb) has been postulated to constitute a prognostic marker for outcome after primary chemoradiation of patients with advanced cancer of the head and neck. However, this hypothesis has not been tested systematically in large study samples. In the years 1992-1997, 125 patients with advanced head and neck cancer (stages III/IV UICC) were treated with primary chemoradiation in two different prospective multicentric trials, 62 patients in trial A (phase II, 1992-1995), and 63 in trial B (phase III, 1995 1997). Beside initial Hb, other pretherapeutic parameters with potential prognostic relevance were assessed and correlated with clinical outcome after 43 months follow-up: total tumor volume (TTV; calculated in initial CT scans), tumor oxygenation (polarographic measurements with Eppendorf histography), TNM, tumor localization, age, and performance status. The evaluation of the clinical end points (progression-free and overall survival and local tumor control) revealed that Hb and TTV were independent parameters with strong predictive character of outcome after primary chemoradiation in both trials (n = 125). Bivariate analysis showed < median (13.5 g/dl) a hazard ratio of 2.1 (P = 0.002) for Hb; and > median (98 ml) a Hazard ratio of 2.0 (P = 0.006) for TTV. Severe anemia (Hb < 10 g/dl) was an adverse factor in three patients. Hypoxia was associated with poorer initial therapeutical response but was not predictive of clinical outcome. Furthermore, tumor oxygenation showed no correlation with Hb. The other parameters examined failed to show prognostic significance. Our results indicate a high prognostic value of initial Hb for outcome after primary chemoradiation in advanced head and neck cancer and imply a therapeutic benefit of Hb substitution or erythropoietin administration. We propose to test this in randomized clinical trials. PMID- 11056854 TI - [Diagnosis and therapy of pseudotumors of the orbits]. AB - Orbital pseudotumor is a nonspecific inflammatory process of unknown etiology that can be divided histopathologically into three basic types: granulomatous, lymphoid, and sclerosing. Between 1995 and 1998, 12 patients with pseudotumor orbitae were treated in the ENT Department of the University of Saarland. Histopathological examination showed granulomatous type of pseudotumor in six, lymphoid in three, and sclerosing in three patients. In seven cases the pseudotumor orbitae were medially located and in four cases laterally. In one patient nearly all orbital structures were infiltrated. Diagnostic biopsy was taken endonasally in six cases, via medical orbitotomy in two cases, and via lateral orbitotomy in four cases. Due to their good delimitation lymphoid and sclerosing tumors were extracted completely during diagnostic biopsy and patients were free of complaints after a few weeks. The six patients with granulomatous pseudotumor were treated primarily with steroids after the diagnosis had been definitely confirmed by histology. In three of those six cases a second course of steroid therapy had to be given, with positive results in two cases. Follow-up was between 6 and 28 months (mean 16 months). There were no postoperative complications. The clinical and radiographic presentation of the pseudotumors can vary greatly. Therefore, the differential diagnosis of specific infections or neoplasms can only be established through diagnostic biopsy. Different rhinosurgical approaches provide clear biopsy results and in some cases the pseudotumor is even completely removed. PMID- 11056855 TI - [Mutational analysis of the connexin26 gene in sporadic cases of moderate to profound deafness]. AB - Non-syndromic neurosensory recessive deafness (NSRD) is one of the most common human sensory disorders. Mutations in the connexin 26 gene have been established as a major cause of inherited and sporadic non-syndromic deafness in different populations. The CX26 gene encodes the gap junction protein connexin 26 (beta-2, GJB2), whose expression was shown in several tissues and in the cochlea. The 30delG mutation is the most frequent mutation in the CX26 gene. It represents a deletion of guanosine (G) in a sequence of six Gs extending from position 30 to 35 of the CX26 cDNA. The deletion creates a frameshift resulting in a premature stop codon and a non-functional intracellular domain in the protein. The 30delG mutation can be detected at the molecular level using PCR followed by BsiYI digestion. We screened 164 mainly German patients with non-syndromic sporadic deafness for this mutation to determine its distribution in the German population. The frequency of the mutation in our analyzed patients was lower than in other studies and therefore indicates its dependency on geographically distinct populations. PMID- 11056856 TI - [Psychological stress, knowledge and treatment expectation of parents with a child managed by cochlear implant]. AB - ESTABLISHED KNOWLEDGE: It is known that parents of hard-of-hearing children suffer from an increase in psychosocial stress. SCIENTIFIC QUESTION: How does the psychosocial situation of parents with children who have cochlear implants change during rehabilitation? AIM OF STUDY: It was the aim of this study to demonstrate how parents evaluate retrospectively their own psychological well-being during the process of rehabilitation. METHODS AND RESULTS: We interviewed 87 parents by questionnaires which were mailed to them. Fifty-seven mothers and 46 fathers responded (59% return rate). Parents reported a significant increase in stress, as perceived by themselves, after the time of diagnosis. Of the parents, 25% continued to suffer from psychic stress during rehabilitation as could be demonstrated by the SCL-90-R questionnaire criteria. The expectations by parents were realistic prior to implantation but thereafter increased significantly with time. CONCLUSIONS: The psychological state of parents during the critical phase, after a diagnosis of deafness has been made for their child, has to be considered. Even after an initial phase of shock, parents seemed to be stressed to an extent that required therapeutic intervention. PMID- 11056857 TI - [Angiosarcoma of the frontal sinus. Case report and review of the literature]. AB - Angiosarcomas are very rare but highly malignant soft tissue tumors derived from the vascular endothelium. The tumor is most commonly found in the skin. The cancer is known to cause early and widespread metastases leading to a very poor prognosis of less than 24 months. The therapy of choice is radical surgery followed by adjuvant radiation. In this case study, we report on a patient with a very unusual localization of angiosarcoma in the frontal sinus. Based on this case, we discuss important aspects of tumor biology, diagnostic procedures, and histologic features as well as therapeutic options. We conclude that angiosarcoma has to be considered by a differential diagnosis in all head and neck neoplasias with uncertain histology. PMID- 11056858 TI - [Enlarged ventricular sacculus in laryngeal carcinoma--in increased risk in tumor surgery]. AB - The case of a 60-year-old female patient with a supraglottic larynx carcinoma (T3N2CM0) and bilateral enlargement of the laryngeal sacculus is presented. On one side, the sacculus opened atypically into the left vestibular fold. The sacculus is a physiologic part of the roof of Morgagni's ventricle, the size of which can vary greatly. These canal-like structures can be the point of origin of cysts, laryngoceles, mucoceles, and pyoceles. Laryngoceles are present in about 5% of larynx carcinomas; their cause is not precisely known. Frequently, a computer tomogram suggests the coincidence of larynx carcinoma and laryngoceles or a large laryngeal sacculus. In oral panendoscopy, these ventricular distensions are difficult to detect even using 70 degrees rigid endoscopes. A planned supraglottic partial resection must take into account early tumor growth along the performed canals in the preepiglottic space. In this patient, a laryngectomy was performed due to the bilateral involvement of the vocal process and the anterior commissure. PMID- 11056859 TI - [Effective and ineffective disinfection measures]. PMID- 11056860 TI - [Fee schedule legislation for physicians--calculation in accordance with standard values]. PMID- 11056861 TI - [Bilateral periorbital edema and right abducens nerve paralysis. Acute right pansinusitis with bilateral cavernous sinus thrombosis]. PMID- 11056862 TI - [Space-occupying lesion of the orohypopharynx. Struma nodosa with retropharyngeal extension]. PMID- 11056863 TI - [Side-effects of nasal CPAP ventilation therapy]. PMID- 11056864 TI - Pediatricians' most difficult decision. PMID- 11056865 TI - From the files of a pediatric ethics committee. PMID- 11056866 TI - Loving noncompliance: determining medical neglect by parents of HIV-positive children. PMID- 11056867 TI - Comment: collaborative decision making with HIV-infected mothers. PMID- 11056868 TI - Responding to parental requests to forego pediatric nutrition and hydration. PMID- 11056869 TI - Futility in pediatrics: from case to policy. PMID- 11056870 TI - Comment: will futility policies make a difference. PMID- 11056871 TI - Ethical issues in pediatric research. PMID- 11056872 TI - Comment: research involving children: clarifying roles and authority. PMID- 11056873 TI - Suffering in children at the end of life: recognizing an ethical duty to palliate. PMID- 11056874 TI - Including the family's interests in medical decision making in pediatrics. PMID- 11056875 TI - Comment: supporting the child within the family. PMID- 11056876 TI - The hazards of "hanging crepe" or stating overly pessimistic prognoses. PMID- 11056877 TI - From case to policy: institutional ethics at a children's hospital. PMID- 11056878 TI - Comment: should all ethics committee members be institutionalized? PMID- 11056879 TI - The antiemetic drug ondansetron interferes with lithium-induced conditioned rejection reactions, but not lithium-induced taste avoidance in rats. AB - Conditioned rejection reactions displayed in the taste reactivity test are exclusively produced by treatments that elicit nausea. The present experiments demonstrate that pretreatment with the antinausea agent ondansetron interferes with both the establishment and the expression of conditioned rejection reactions. Ondansetron did not interfere with lithium-induced taste avoidance in either a 1-bottle or a 2-bottle test. In fact, when rejection reactions were measured during a consumption test, ondansetron selectively attenuated rejection reactions, with only a slight modification of consumption. These results suggest that conditioned rejection reactions, but not conditioned taste avoidance, reflect nausea in rats that can be attenuated by ondansetron pretreatment. PMID- 11056880 TI - Time course of cardiac conditioned responses in restrained rats as a function of the trace CS-US interval. AB - Two experiments aimed at understanding the temporal characteristics of trace conditioned heart-rate responses to a 0.5-s tone (conditioned stimulus [CS]) in restrained rats. A CS paired with a tail-shock (unconditioned stimulus [US]) elicited lasting bradycardiac responses. The magnitude and extinction rate of conditioned responses (CR) were independent of the CS-US interval (interstimulus interval [ISI], 3 s to 20 s). Unreinforced test trials were analyzed for CR topography. Responding was delayed in groups with longer ISIs, but CR latencies, peak and decay times were not proportional to the ISI. Response peaks tended to cluster either about 6 s after CS onset, or about 10 s with a slow decay, depending on the ISI. The authors postulated 2 components of auditory stimulus traces involved in cardiac conditioning, maximally active 6 s and 10 s respectively after CS onset. The topography of the CR could be constrained to combinations of associative strength and instantaneous activity of these 2 components. PMID- 11056881 TI - Can rats acquire an olfactory learning set? AB - Experimental rats were trained on multiple 2-odor discrimination tasks, whereas controls were given repeated sessions on Task 1 and then were tested on a novel 2 odor task. Experimental rats showed strong positive transfer across problems and approached errorless or near-errorless learning. Control rats maintained near perfect performance on Task 1 but performed at chance on initial trials when tested with novel odors. Thus, the near-errorless terminal performance of experimental rats was a function their having been trained on multiple problems and was not simply the result of eliminating "disruptive response tendencies" (I. C. Reid & R. G. M. Morris, 1992). Results support the view that when rats are trained on a series of 2-odor discrimination tasks, they acquire a strategy or rule that allows them to solve new problems with few or no errors. PMID- 11056882 TI - Free-operant performance on variable interval schedules with a linear feedback loop: no evidence for molar sensitivities in rats. AB - Four experiments examined rats' sensitivity to molar and molecular factors on instrumental schedules of reinforcement. Rats were exposed to a variable interval schedule with a positive feedback loop (VI+), such that faster responding led to a shorter interreinforcement interval. In Experiments 1 and 2, rats responded faster on a variable response (VR) schedule than on either a VI schedule matched for reinforcement rate or a VI+ schedule matched for the feedback function. In Experiment 3, rats responded no differently on a VI schedule than they did on a VI+ schedule with equated rates of reinforcement. In Experiment 4, rats responded faster on a VI+ schedule with an interresponse time requirement yoked to that experienced on a VR schedule, than on a VI+ schedule with the same feedback function as the VR schedule. Taken together these results suggest that rats are more sensitive to the molecular than the molar properties of the schedules. PMID- 11056883 TI - Associative changes in excitors and inhibitors differ when they are conditioned in compound. AB - When an AB stimulus compound is reinforced or nonreinforced, there are associative changes in both A and B elements. In many contemporary theories those changes are viewed as governed by a common error term, computed as the discrepancy between the total associative strength of the AB compound and that supported by the trial consequence. This implies that if A and B are equally salient, then the magnitude of their associative change should be the same, whatever their strengths prior to the AB trial. This implication was explored for a compound consisting of an excitatory A and an inhibitory B. A novel assessment procedure avoided the difficulty of making comparisons at different locations on the performance scale. Three experiments using a magazine approach preparation in rats and 3 using autoshaping in pigeons found evidence contradicting this implication. The excitatory A changed less than the inhibitory B when the compound was reinforced but more than B when the compound was nonreinforced. PMID- 11056884 TI - Geometric rule learning by Clark's nutcrackers (Nucifraga columbiana). AB - Clark's nutcrackers (Nucifraga columbiana) were trained to search in a location defined by its geometric relationship to 2 landmarks. Two groups were trained to search at different points along the line connecting the landmarks, and 2 groups were trained to find the 3rd point of a triangle, on the basis of either direction or distance from the landmarks. All groups learned and transferred to new interlandmark distances. However, the constant-distance group learned more slowly, searched less accurately, and showed less transfer than the other 3 groups. When tested with new orientations of the landmarks, the birds tended to follow small but not large rotations. When tested with a single landmark, birds in the half, quarter, and constant-bearing groups searched in the appropriate direction from the landmark, but birds in the distance group did not. These results demonstrate that nutcrackers can learn a variety of geometric principles, that directional information may be weighted more heavily than distance information, and that the birds can use both absolute and relative information about spatial relationships. PMID- 11056885 TI - Conditioning across the duration of a backward conditioned stimulus. AB - Five conditioned suppression experiments examined the extent to which an appetitively motivated lever-press response can be punished by different components of a backward conditioned stimulus (CS). Using a 0-s unconditioned stimulus (US)-CS interval, Experiments 1 and 2 showed that the initial 3 s of a normally 30-s backward CS served as a more effective punisher than the CS as a whole. Experiment 3 found no such effect if the US-CS interval were 3 s rather than 0 s. Experiments 4A and 4B found that if the US-CS interval were 0 s, the initial part of the backward CS acquired excitatory properties although the CS as a whole passed a summation test for conditioned inhibition. By contrast, the 3-s US-CS interval supported inhibitory conditioning across the whole duration of the backward CS. Taken together, these findings support a modified version of Wagner's sometimes opponent process model, which suggests that different components of a backward CS become either excitatory or inhibitory depending on the components' temporal proximity to the US. PMID- 11056886 TI - The influence of associability changes in negative patterning and other discriminations. AB - Normal rats showed faster learning of a serial negative patterning (NP) discrimination (X+, A+, X-->A-) than of a comparable feature negative (FN) discrimination (A+, X-->A-). This advantage was absent in rats with lesions of the amygdala central nucleus. Earlier data indicated that this brain lesion interferes with surprise-induced increases in attention specified by the Pearce Hall model (J. M. Pearce & G. Hall, 1980). In the NP task, but not the FN task, omission of the reinforcer after X on X-->A- trials was surprising. A variation of the NP task (NPX), in which X was reinforced on both X+ and X-->A- trials, was learned more rapidly than the NP task. Lesioned rats were unimpaired in learning the NPX task. Evaluation of the lesion effects and the results of posttraining transfer tests suggested that the NP advantage involved attentional processes, whereas the NPX advantage was based on the acquisition of inhibitory control by aspects of excitation conditioned to X. PMID- 11056887 TI - Searching for the center: spatial cognition in the domestic chick (Gallus gallus). AB - Chicks learned to find food hidden under sawdust by ground-scratching in the central position of the floor of a closed arena. When tested inan arena of identical shape but a larger area, chicks searched at 2 different locations, one corresponding to the correct distance (i.e., center) in the smaller (training) arena and the other to the actual center of the test arena. When tested in an arena of the same shape but a smaller area, chicks searched in the center of it. These results suggest that chicks are able to encode information on the absolute and relative distance of the food from the walls of the arena. After training in the presence of a landmark located at the center of the arena, animals searched at the center even after the removal of the landmark. Marked changes in the height of the walls of the arena produced some displacement in searching behavior, suggesting that chicks used the angular size of the walls to estimate distances. PMID- 11056888 TI - Parents accuse APA of Ritalin conspiracy? PMID- 11056889 TI - Validating behaviors in Alzheimer's disease patients. PMID- 11056890 TI - Decrease in restraint use in a study of a geropsychiatric unit. AB - 1. Traditional and nontraditional interventions were used in this study. 2. There was a reduction in the number of times restraints were used, as well as in the number of patient placed in restraints. 3. Further research is needed to find more humane ways to help older adults through acute crisis episodes. PMID- 11056891 TI - The astronaut's wife. Lessons of growth through spousal depression. AB - 1. Living with cyclical spousal depression presents both challenges and opportunities for personal growth. 2. Denial can be a significant barrier to the treatment of depression. 3. Spouses of chronically depressed individuals also are at risk for depression. A preventive family-centered approach should be used. PMID- 11056892 TI - The history, economics, and financing of mental health care. Part 2: The 20th century. AB - 1. To understand how mental health nursing practice was affected by the financing and policy changes occurring rapidly in the second part of the 20th century, sources can only be found in the literature in psychiatry, the social sciences, and economics. There was no psychiatric nursing journal until the 1950s, and no article by a nurse in the general nursing literature about finances. 2. Deinstitutionalization was really transinstitutionalization. Changes in regulations in Medicaid allowed the shifting of mentally ill people who were older than age 65 to nursing homes. 3. Community mental health centers never developed programs to serve people who were seriously mentally ill. Rather than serving clients who were psychotic, the community mental health centers marketed their treatment programs to people with anxieties, who were undergoing divorce, or who had mildly troubled children. PMID- 11056893 TI - Flight to Los Angeles. Crisis at 30,000 feet. AB - 1. Using therapeutic communication, the nurse can begin to assess the individual in crisis, establish rapport, and evaluate the crisis situation. 2. Offering self while remaining calm assists in de-escalating the anxiety felt by an individual in crisis. 3. Communications with others at the crisis situation, as well as the individual in crisis, should be direct, calm, clear, and incorporate safety assessments as the situation unfolds. Assuring everyone that the nurse is there to help enlists cooperation in crisis resolution. PMID- 11056894 TI - Portrait of a person with schizophrenia. PMID- 11056895 TI - Premature ejaculation: a psychophysiological approach for assessment and management. AB - This article distinguishes several subtypes of biogenic and psychogenic premature ejaculation (PE) according to their etiologic features: the physiological PE types of (a) neurologic constitution, (b) acute physical illness, (c) physical injury, and (d) pharmacologic side effect; and the psychological PE types of (a) psychological constitution, (b) acute psychological distress, (c) relationship distress, and (d) psychosexual skills deficit. Attention is given to assessment and differential diagnosis, and to specific treatment of the types of PE, such as the pharmacologic management of difficult neurologic cases. Effective psychosexual treatment combines multiple strategies such as physiological relaxation, pubococcygeal muscle training, cognitive and behavioral pacing strategies, and the involvement of the partner in the therapy. Treatment should determine the specific type of PE and comprehensively address its particular features in order to improve long-term treatment effectiveness. PMID- 11056896 TI - Conflict management style and marital satisfaction. AB - The aim of this study was to investigate whether there is one conflict management style that correlated more significantly with marital satisfaction than any other. In addition, spousal satisfaction with how marital conflict is managed was also examined, as were gender differences. Fifty-seven couples who had been married for at least 10 years took part in the study. Results showed that the collaborative conflict management style has the highest correlation with both marital satisfaction and spousal satisfaction with conflict management in the marriage. In contrast, where one or both of the spouses used the competitive conflict management style, the lowest marital satisfaction was reported. The results were also interpreted in terms of cultural and gender differences. PMID- 11056897 TI - A homemade device for erectile dysfunction. PMID- 11056898 TI - Amputee fetishism and genital mutilation: case report and literature review. AB - A case is presented of a 49-year-old man who amputated his penis following instructions that he had obtained from the Internet. The patient had a long standing amputee fetish, which evolved into eroticized genital mutilation. The transformation of the preferred fetish occurred in a setting of depression due to environmental stressors. The literature about amputee fetishism, also called "apotemnophilia," is reviewed, and possible connections with the genital mutilation are discussed. PMID- 11056899 TI - A clinical and psycho-sociological case study on gender identity disorder in Japan. AB - Gender identity disorder (GID) has been attracting increasing attention in recent years in Japan as well as in the rest of the world. This report presents a GID case followed by a discussion on GID patients in Japanese culture, social pressures upon diagnosis and treatment of GID, and social changes that have brought about the possibility for GID diagnosis and treatment. PMID- 11056900 TI - Ciprofloxacin as cure of premature ejaculation. AB - This article presents the case report of a 31-year-old patient of mine with both an inflamed prostate gland and, unknown to his doctor, premature ejaculation, both of which were treated successfully by the antibiotic ciprofloxacin. PMID- 11056901 TI - Suction to the breasts of a transsexual male. AB - Patients attending gender dysphoria clinics may report attempts to hasten the transition to the preferred gender. This may involve ingestion of higher quantities of hormones than prescribed or acts of self-mutilation to the genitals. A case is reported of an attempt to encourage breast-tissue development using a sink plunger. PMID- 11056902 TI - Sildenafil for St. John Wort-induced sexual dysfunction. PMID- 11056903 TI - Use of lithium and depot neuroleptics in the treatment of paraphilias. PMID- 11056904 TI - Lithium for paraphilias? Probably not. PMID- 11056905 TI - Custom-coloring your EH&S program. PMID- 11056906 TI - Getting more from your OHS software system. PMID- 11056907 TI - Who wants to be a safety millionaire? PMID- 11056908 TI - Effective motivation. PMID- 11056909 TI - Making safety fun. PMID- 11056910 TI - Dual protection. PMID- 11056911 TI - Hearing aids in occupational settings: safety and management issues. PMID- 11056912 TI - Best foot forward. PMID- 11056913 TI - Market-driven makeover. PMID- 11056914 TI - Getting a grip: the flat dip revolution. PMID- 11056915 TI - Building a better garment. PMID- 11056916 TI - The right stuff. PMID- 11056917 TI - A proactive approach. PMID- 11056918 TI - Business continuity planning. PMID- 11056919 TI - Focused on integrated administration. AB - As employers continue to tinker with designing stronger benefits packages, the impetus to reevaluate once-overlooked areas such as how their statutory disability program is doing probably will get more notice, as well. For employers residing in states with government-operated and mandated disability programs, the option at least to explore self-insuring this program should not be overlooked. PMID- 11056920 TI - Computer software for process hazards analysis. AB - Computerized software tools are assuming major significance in conducting HAZOPs. This is because they have the potential to offer better online presentations and performance to HAZOP teams, as well as better documentation and downstream tracking. The chances of something being "missed" are greatly reduced. We know, only too well, that HAZOP sessions can be like the industrial equivalent of a trip to the dentist. Sessions can (and usually do) become arduous and painstaking. To make the process easier for all those involved, we need all the help computerized software can provide. In this paper I have outlined the challenges addressed in the production of Windows software for performing HAZOP and other forms of PHA. The object is to produce more "intelligent", more user friendly software for performing HAZOP where technical interaction between team members is of key significance. HAZOP techniques, having already proven themselves, are extending into the field of computer control and human error. This makes further demands on HAZOP software and emphasizes its importance. PMID- 11056921 TI - Weighing MSDS management programs. PMID- 11056922 TI - For safety light curtains, location is everything. PMID- 11056924 TI - Computer workstation ergonomics guidelines. PMID- 11056923 TI - Modern methods. PMID- 11056925 TI - Real relief. PMID- 11056926 TI - Improving productivity in tool-intensive operations. PMID- 11056927 TI - A big case for employers. PMID- 11056928 TI - Keys to a successful program. AB - Drug testing has proven to be an effective safety strategy. With proper planning, implementation, and technical expertise, a company can enjoy the benefits of a drug-free workplace. Should you wish to outsource this responsibility, there are some very good Program Administrators throughout the country, some national in scope and some regional. When recruiting an administrator, make sure it is capable of satisfying your needs. These may include a national collection site network with one fee for all your locations and with appointment scheduling, Medical Review Officers on staff with 24-hour availability, custom or standard policy and procedure manuals, training capabilities, the computing ability to customize your program, and the references to back up what the administrator claims. (Fact: If you don't test for drugs and all other employers in your area do, where do you think the drug users work?) PMID- 11056929 TI - Training to second nature. PMID- 11056930 TI - Emphasizing job site safety. PMID- 11056931 TI - The lowdown on lighting. PMID- 11056932 TI - A pediatrician's view. A brief history of otitis media. PMID- 11056933 TI - Otitis media: what do we know at the start of the new millennium? PMID- 11056934 TI - Acute otitis media: diagnosis and management in the year 2000. PMID- 11056935 TI - Otitis media with effusion in children: a pediatric office perspective. PMID- 11056936 TI - Otitis media: the otolaryngologist's perspective. PMID- 11056937 TI - Complications of otitis media. PMID- 11056938 TI - Vaccines and otitis media. PMID- 11056939 TI - Resident's column. PMID- 11056940 TI - Ending violence against women. PMID- 11056941 TI - The deafness of Beethoven: a paradigm of hearing problems. PMID- 11056942 TI - Dissociation of morphine-induced cyclic AMP-upregulation and mu-opioid receptor desensitization. PMID- 11056943 TI - Peripheral antinociceptive interaction of dipyrone and morphine in the formalin test. PMID- 11056944 TI - Mu opioid analgesia and analgesic tolerance in two mouse strains: C57BL/6 and 129/SvJ. PMID- 11056945 TI - Influence of pregnancy on cardiovascular responses to serotonin. PMID- 11056946 TI - Subrenal aorta coarctation of pregnant rats: changes in vascular contractility. PMID- 11056947 TI - Ammonia effects on ionotropic glutamate receptors. PMID- 11056948 TI - GABA binding to cerebral cortical GABAA and GABAB sites in aging mice. PMID- 11056949 TI - Fungi test for evaluation of new antiandrogenic molecule. AB - Antiandrogens have high impact in medicine as they are compounds capable of decreasing the effect of benign prostatic hyperplasia, cancer, and other androgen dependent diseases that affect a large percentage of the male population in the world. [figure: see text] Dihydrotestosterone, a 5 alpha-reductase metabolite of testosterone, has been implicated as the responsible factor of these androgen abnormalities. This fact indicates that the logical step in the treatment of these diseases is the inhibition of this enzyme activity using molecules that compete selectively with its natural substrates. The study of the pharmacological effect of antiandrogens requires specific animal models using a large number of laboratory animals. On basis of this concept, presently there is a tendency to eliminate studies with animals. In this paper we report a new method for antiandrogenic evaluation using microbial transformations. PMID- 11056950 TI - Antiandrogenic effect of new synthetic steroids. AB - These results indicate that castration decreased the flank organ diameter and the weight of the seminal vesicles, whereas daily injection of testosterone restores them. However injections of finasteride together with testosterone inhibited the diameter of the flank organs, the weight of the seminal vesicles, and the conversion of testosterone to dihydrotestosterone. The four compounds tested inhibited flank organ diameter size and the weight of the seminal vesicles as compared to the testosterone-treated animals. On the other hand, none of these compounds suppressed the conversion of testosterone to dihydrotestosterone. These results suggest that the synthesized steroids compete with androgen receptors. PMID- 11056951 TI - Dose-related atropine-induced insulin resistance: comparing intraportal versus intravenous administration. PMID- 11056952 TI - Effects of oxcarbazepine on the behavioral response and neuroanatomical alterations following administration of kainic acid. PMID- 11056953 TI - Monolysocardiolipin acyltransferase activity: effect of insulin and localization of activity. PMID- 11056954 TI - Effects of isoproterenol administered during the last third of gestation on the productive parameters of primiparous sows. PMID- 11056955 TI - U-50,488H, a kappa opioid receptor agonist, is a more potent blocker of cardiac sodium channels than lidocaine. PMID- 11056956 TI - Effect of dexamethasone plus vitamin B complex in the PIFIR model. PMID- 11056957 TI - Ubiquinone (coenzyme Q10) and complex I in mitochondrial oxidative disorder of Parkinson's disease. PMID- 11056958 TI - Efficacy of cefixime in the therapy of typhoid fever. PMID- 11056959 TI - Effect of glycine on plasma levels of glucose and insulin in healthy volunteers. PMID- 11056960 TI - Dexamethasone alone versus dexamethasone plus complex B vitamins in the therapy of low back pain. PMID- 11056961 TI - Novel inhibitors of botulinus neurotoxin "A" based on variations of the SNARE motif. PMID- 11056962 TI - Evidence that simulated microgravity may alter the vascular nonreceptor tyrosine kinase second messenger pathway. AB - Simulated microgravity (hind limb unweighting; HU) reduces maximal contractile capacity to norepinephrine (NE) but not 5-hydroxytryptamine (5-HT) in the rat abdominal aorta of male Wistar rats. Our earlier study showed that voltage operated calcium channels, the MAPK pathway [1], and vasoconstrictive prostaglandins contribute to the NE-induced contraction of control (C) but not HU, aorta rings. Genistein, a general tyrosine kinase inhibitor, caused a significant reduction in vascular contractility in C but not HU arteries. The present study explored the role of protein kinase C (PKC) and extracellular receptor-activated kinase 1 and 2 (ERK1/2) in the HU-induced vascular hyporesponsiveness to NE. Microgravity was simulated in Wistar rats by 20 day HU. The abdominal aorta was removed from control and HU rats, cut into 3 mm rings, and mounted in tissue baths to measure isometric contraction. Protein levels were determined using Western blot analysis. PD98059, a selective MAPKK inhibitor, caused a marked inhibition of NE-induced contraction in both C and HU arteries. Calphostin C, a PKC inhibitor, completely abolished the contractile response to NE in both C and HU tissues. Phosphorylated (activated) ERK1/2 protein mass was greater in C, compared to HU, aortas, and was reduced by genistein only in C tissues. MAPK total protein levels in the rat aorta were increased in the HU treated, compared to C, animals. These results indicate that PKC represents an early transduction step in the contractile response to NE in the rat abdominal aorta. That inhibition of the step immediately before activation of MAPK reduced contraction in both C and HU tissues, while general tyrosine kinase inhibition with genistein blocked only the control responses, suggests that a nonreceptor tyrosine kinase may be involved in HU-induced vascular hyporesponsiveness to NE. PMID- 11056963 TI - The effect of first and second beer consumption on breathalyzer results. PMID- 11056964 TI - Prenatal ethanol exposure results in morphometrical alterations of immunoreactive neurons to vasopressin in the suprachiasmatic nucleus of young rats. PMID- 11056965 TI - Ginkgo biloba pretreatment partially protects from the dopaminergic neurotoxicity of 1-methyl-4-phenylpyridinium. PMID- 11056966 TI - A new therapeutic scheme of ibuprofen to treat postoperatory endodontic dental pain. PMID- 11056967 TI - Publishing pharmacology in the twenty-first century. AB - The World Wide Web and other forms of digital communication have lead some to predict the demise of printed journals. In my experience, scientific articles can be more efficiently reviewed and edited by digital document sharing. High quality print journals will remain the preferred scholarly venue for authors' best works, but only if the ethical behavior of scientists can be maintained and appropriately rewarded. PMID- 11056968 TI - Two hundred years of pharmacology: a midpoint assessment. PMID- 11056969 TI - [MR angiography of the supraaortic vessels]. AB - PURPOSE: To compare high resolution contrast-enhanced MR angiography (MRA) and digital subtraction angiography (DSA) in the assessment of supraaortic vessel stenosis. METHODS: 14 patients with suspicion of cerebrovascular disease or upper limb ischemia underwent selective DSA and high resolution contrast enhanced MRA employing a new Panoramic-Array coil. Stenosis assessment in comparison to DSA followed NASCET criteria. Additionally signal-/noise ratios (SNR) were evaluated to assess contrast enhancement. RESULTS: Diagnostic image quality was achieved in all patients. Sensitivity and specificity for assessing high-grade stenosis of the supraaortic vessels were 100% and 96% respectively. In the assessment of high grade common or internal carotid artery stenosis sensitivity and specificity was 100%. CONCLUSION: High resolution contrast enhanced supraaortic MRA combined with new coil systems allow for a reliable assessment of stenoses along the whole vessel course including the aortic arch. Previous stent procedures limit its use in postinterventional follow-up. PMID- 11056970 TI - [The indications and embolism monitoring in lumen-opening therapies of the a. carotis]. AB - Carotid endarterectomy (CEA) is proven to be beneficial in symptomatic patients with high-grade carotid stenosis (70% to 99%; residual lumen as a percentage of the normal distal internal carotid artery) on condition that the peri-operative risk for mortality and morbidity is less than 6%. A minority of the "leading experts" in North America (48%) and Western Europe (28%) recommends carotid endarterectomy in asymptomatic patients in general. Most experts suggest to perform surgery only in asymptomatic patients who are at risk for carotid occlusion in the near future or embolism. At its present state, angioplasty and stenting is an experimental although promising technique which will have to be compared to carotid endarterectomy. Criteria for duplex grading of internal carotid stenosis have been established and systematically validated to results of angiography. Pre-surgical use of angiography will more and more be restricted to selected patients in whom the results of duplex sonography remain inconclusive. The detection of microemboli with transcranial doppler sonography seems to be of particular importance before and during carotid angioplasty and stenting. PMID- 11056971 TI - [Carotid stenosis: the technic for PTA and stent implantation]. AB - Percutaneous transluminal angioplasty and stenting of carotid artery stenosis represents an innovative experimental modality which has been increasingly employed in high-risk patients. Currently, results from prospective randomized trials are not available; thus, guidelines with regard to indications and technique are preliminary. Numerous technical innovations to avoid associated complications have evolved in neurointerventional procedures. Dedicated guiding catheters and stents using a transfemoral approach have been introduced for treatment of carotid artery disease. Low-profile stent delivery systems may decrease risk of releasing embolic load when crossing high-grade stenosis or kinking of the vessel. The introduction of a cerebral protection with the use of temporary occlusion balloons or filter systems has resulted in a reduction of procedure-related neurological complications, and is strongly recommended by leading experts in the field. PMID- 11056972 TI - [Stenoses and occlusions of the supraaortic vessels--the interdisciplinary interventional treatment of patients with an increased surgical risk]. AB - PURPOSE: The value of elective PTA in stenoses of the supraaortic vessels and especially of the carotid artery is still under discussion. Stenoses after surgical and/or radiation therapy and a high cardio-vascular risk for a surgical procedure, however, justify minimal-invasive techniques. MATERIAL AND METHODS: In 31 patients with 24 symptomatic and 7 asymptomatic stenoses of the supraaortic arteries (3 Acc, 19 Aci, 5 Ascl, 4 Tr.brc, 2 Aax) minimal-invasive angioplasty and stent-placement was performed. The rationale for the interventional procedure was an elevated medical or technical risk of the respective surgical procedure. All procedures were performed using a coaxial technique with a guiding catheter. An open vascular access was necessary in 7 cases due to an unfavorable anatomical situation. RESULTS: The technical success rate was 100%. Peri-interventionally, a TIA occurred in 2 patients (1 spontaneous relieve, 1 complete remission after rt PA lysis therapy). During the mean follow-up period of 13 months 3 patients died due to non-cerebrovascular reasons. In 2 patients a moderate restenosis, in 1 a high-grade stenosis, and in 1 and complete occlusion occurred without symptoms. After one year of follow-up, all treated stenoses were patent with a restenosis rate of 10.7% without further neurological symptoms. CONCLUSION: Interventional angioplasty with stent-placement of the supraaortic arteries is a safe and effective alternative in patients are at high risk for classical surgical procedures. PMID- 11056973 TI - [Carotid endarterectomy and carotid stenting. A pilot study of a prospective, randomized and controlled comparison]. AB - INTRODUCTION: A prospective, randomized and controlled trial is conducted to compare carotid endarterectomy and carotid stenting in high grade symptomatic carotid artery stenoses. METHODS: According to the study design symptomatic patients with a angiographically high-grade (> or = 70%) internal carotid artery stenosis are included. Pre- and postinterventional diagnostics during the hospitalization period includes neurological assessment, duplex sonography of the cervical and cerebral arteries and magnetic resonance imaging of the brain. Follow-up examinations are scheduled after 1, 6 and 12 months and consist of a neurological assessment and duplex sonography. After 12 months selective angiography and magnetic resonance imaging of the brain will be performed additionally. During a period of 9 months up to now 23/137 patients treated for a carotid artery stenosis were included in the study, 11 patients underwent surgery and 12 patients carotid stenting. RESULTS: Carotid stenting and endarterectomy was primarily successful without residual stenosis > 30% in each patient without the occurrence of stroke or death. In 18 follow-up examinations (neurological assessment including duplex sonography) of 13 patients (13 follow-up examinations after 30 days, 5 after 6 months) no relevant restenosis and no stroke occurred. CONCLUSION: As of yet, carotid stenting was a safe procedure. Due to the small number of patients a definitive conclusion can not be drawn. PMID- 11056974 TI - [Reconstructive endovascular treatment procedures in the area of the a. subclavia and its branches]. AB - PURPOSE: Principles, methods and results of percutaneous therapy in acute and chronic vascular alterations of the subclavian artery and its branches. MATERIAL AND METHODS: 17 arteries in 15 patients have been treated. The patients suffered from stenoses (n = 6), occlusions (n = 7), aneurysms, (n = 2) or acute hemorrhage (n = 2). The lesions were located in the subclavian artery (n = 8), the axillary artery (n = 4) or the vertebral artery (n = 5). In all cases the procedures were performed via via a transfemoral approach. Balloon angioplasty was employed in 13 cases, in 6 cases bare stents and in 4 cases grafted stents were implanted. In one case additionally local lysis was performed. RESULTS: Primary technical success was achieved in all cases. No catheter complications did occur. In 13 Patients complete follow-up examinations could be performed. The life-threatening acute hemorrhages were stopped without recurrent bleeding. The aneurysms could be completely excluded without endoleaks. In 4 Patients of the stenosis-group re stenosis occurred and following stent-implantation one occlusion after 6 months was observed. 8 patients are without clinical symptoms since the intervention. CONCLUSION: Stenotic and occlusive vascular alterations as well as aneurysms and the acute hemorrhage in supraaortic arteries can be effectively and safely treated by endovascular techniques using percutaneous groin access. PMID- 11056975 TI - [The diagnosis of metastases in the bone marrow by MRT]. AB - MRI plays an important role in the diagnostic workup of skeletal metastases. In principle two different applications of MRI can be distinguished: apart from the well known indications of clarifying uncertain lesions seen with other imaging modalities or demonstrating known osseous lesions with high resolution imaging for therapeutic planing, MRI can also be used as a primary screening modality for skeletal metastases. Besides the higher lesion detection rate the major advantage of MRI compared to bone scintigraphy lies in the demonstration of morphology which on the one hand exactly shows the extension of tumorous lesions and on the other hand clearly distinguishes between malign and benign processes. As unclear findings on bone scintigraphy often require additional imaging studies, especially in patients with clinical findings or lab results suggestive for metastatic disease, we think that whole-body-skeletal-MRI is not only an accurate but also a cost effective diagnostic modality in the detection and screening for skeletal metastases, if the indication for the examination is closely related to clinical findings and the therapeutic relevance of the imaging results. PMID- 11056976 TI - [Breast cancer screening. The paper by J. Engel et al. "The early detection of breast cancer in Germany. The time to deal with it". Radiolge (2000) 40:177-183]. PMID- 11056977 TI - [Chronic recurring ileal states]. PMID- 11056978 TI - [Good clinical practice. Its meaning for clinical research]. PMID- 11056979 TI - Spurious species? PMID- 11056980 TI - All doped up--and going for the gold. PMID- 11056981 TI - What's the matter? The prevailing theory for the universe's "missing mass" stumbles. PMID- 11056982 TI - For the bees. Glowing paint may highlight the forces that make insects fly. PMID- 11056983 TI - Three-star performance. Tomography from the ground could outdo the hubble and its successor. PMID- 11056984 TI - A new rex. The biggest meat eater of them all bolsters the theory of pack hunting. PMID- 11056986 TI - The biologist and the cathedral. PMID- 11056985 TI - Physician, heal thyself. PMID- 11056988 TI - DVDs: cease and DeCSS? PMID- 11056987 TI - Wired for speed. As chips shrink, researchers look to optical and radio-frequency interconnects. PMID- 11056989 TI - The small planets. PMID- 11056990 TI - Avoiding a data crunch. PMID- 11056991 TI - Coping with crowding. PMID- 11056993 TI - Self-service stations_gas nozzles. Fill 'er up. PMID- 11056992 TI - Making metallic hydrogen. PMID- 11056994 TI - Care for a dying continent. PMID- 11056995 TI - Fun with flat fluids. PMID- 11056996 TI - Rep-tiling the plane. PMID- 11056997 TI - Netting the deep sky. PMID- 11056998 TI - What a nerve. PMID- 11056999 TI - The additivity of stimulus-response compatibility with perceptual and motor factors in a visual choice reaction time task. AB - One hundred and four participants performed a two-choice reaction time task requiring button-press responses to visual stimuli. Two levels of stimulus response compatibility were factorially combined with two levels of stimulus discriminability and two levels of response repertoire. Results showed that the effects of these three variables were additive both in terms of mean reaction time as in terms of reaction time variances. The implication of this outcome is discussed in terms of the underlying information processing stage structure. PMID- 11057000 TI - Mechanical perturbation of the wrist during one-handed catching. AB - In the present study, the co-ordination of grasp and transport components of one handed catching was examined following mechanical perturbations applied to the wrist. Six skilled catchers (mean age = 27.5 years) performed 64 trials in which tennis balls were projected at approximately 8 ms-1. The trial blocks consisted of 10 non-perturbed trials (NPTs) (baseline), and a block of 54 trials of which 20 trials were perturbed. The perturbation was in the form of a resistive force (12 N) applied via a piece of cord attached to a mechanical brake. In baseline trials participants reached maximal wrist velocity closer to the time of hand ball contact (237 ms +/- 68) than in the perturbed (309 ms +/- 61) condition. Furthermore the wrist velocity profile of five out of six participants exhibited a double peak immediately after a perturbation. However, aperture variables such as the relative moment of final hand closure (approximately 70% of overall movement time) were not typically affected. The stability of grasp and transport coupling for one-handed catching was shown to vary from trial to trial. Skilled performers exploited redundant degrees of freedom in the motor system when faced with a sudden, unexpected change in task constraints. PMID- 11057001 TI - Sentence reading: do we make use of orthographic cues in homophones? AB - Starting from the finding that currently phonological models of visual word processing predominate, we examined what happened when important morphological information is disclosed in the orthography but not in the phonology. To do so, we made use of a peculiarity in Dutch. In this language, some forms of the present and the past tense of verbs are homophones or homographs. This allowed us to look at the power of orthographic and phonological cues to derive the tense of the verb. Two experiments showed that orthographic cues alone suffice to recover the tense of the verb, and that this recovery does not take more time than tense recovery on the basis of a combination of orthographic and phonological cues. On the basis of these results, we conclude that orthographic cues in homophones are very efficient during silent reading. Our findings, however, do not allow us to conclude whether this is due to a direct route from orthography to meaning, or to a specialised, morpho-syntactic back-up strategy elicited by certain sequences of letters. PMID- 11057002 TI - Evidence of imagined passive self-motion through imagery-perception interaction. AB - The existence of whole-body passive self-motion mental imagery was investigated by examining whether the perception of passive body accelerations can be affected by passive self-motion imagery. Twenty healthy subjects recognised target passive body acceleration. This recognition task was performed under three conditions: (1) a baseline condition without imagery; (2) a compatible imagery condition during which subjects imagined themselves passively moving in the same direction as the target acceleration; (3) a non-compatible imagery condition during which subjects imagined themselves passively moving in the direction opposite to that of the target acceleration. The recognition of the target acceleration was improved under compatible and degraded under non-compatible imagery. This interaction implies that perception and imaginary share common representations, and supports the existence of passive self-motion imagery. PMID- 11057004 TI - Inhibitory motor control in stop paradigms: comment on Band and van Boxtel (1999). PMID- 11057005 TI - The influence of time pressure and cue validity on response force in an S1-S2 paradigm. AB - Hypotheses about variations of response force have emphasised the influences of arousal and of motor preparation. To study both types of influences in one experiment, the effects of time pressure and of validity of S1 were investigated in tasks wherein a first stimulus (S1) indicated the most probable response (80% valid) required after a second stimulus (S2). Under time pressure, responses were executed more forcefully while, as could be expected, response times were shorter and errors were more frequent. This pattern of results was not only obtained when time pressure was varied between blocks, but also when varied from trial to trial, by information given by S2. Also invalidly cued responses were executed more forcefully but, as could be expected, in contrast to time pressure, response times were longer and errors were more frequent. The results demonstrate that latency and force of responses may vary in different directions. Ways are outlined on how current hypotheses must be extended in order to account for these results. PMID- 11057006 TI - Contour symmetry detection: the influence of axis orientation and number of objects. AB - Participants discriminated symmetrical from random contours connected by straight lines to form part of one- or two-objects. In experiment one, symmetrical contours were translated or reflected and presented at vertical, horizontal, and oblique axis orientations with orientation constant within blocks. Translated two object contours were detected more easily than one, replicating a "lock-and-key" effect obtained previously for vertical orientations only [M. Bertamini, J.D. Friedenberg, M. Kubovy, Acta Psychologica, 95 (1997) 119-140]. A second experiment extended these results to a wider variety of axis orientations under mixed block conditions. The pattern of performance for translation and reflection at different orientations corresponded in both experiments, suggesting that orientation is processed similarly in the detection of these symmetries. PMID- 11057007 TI - Hepatitis A in the northern interior of British Columbia: an outbreak among members of a first nations community. PMID- 11057008 TI - Case-control study assessing the association between yersiniosis and exposure to salami. PMID- 11057009 TI - [MR imaging of the subcapital growth plate of the proximal femur]. AB - The possibilities of MR imaging of the subcapital growth plate of the proximal femur were evaluated in 36 children with 58 normal hips. The most common presentation of the area of subcapital growth plate on MR images was that of a hypointense band on T1-weighted images and a hyperintense band on sequences using fat saturation. Thinning of the growth plate with age was observed and in the older subjects in this group the growth plate was no longer visible. PMID- 11057010 TI - [Clinical, radiological and personal condition of patients over 18 year of age with congenital hip dislocation treated conservatively]. AB - The paper presents the clinical, radiological and personal condition of 121 patients (162 hip joints) age 18 to 32 years (mean age 24 years) treated during childhood because of congenital hip dislocation. In 34% of the cases slight and moderate pain was reported, but no other symptoms were found during follow-up. Radiological condition was assessed as very good in 34% of the cases, good in 48%, satisfactory in 6% and unsatisfactory in 5%. Structural changes in spongy bone, which may be early signs of degenerative disease were found in 3 hips. These changes were visible on X-ray films as a sign of increased density in the upper rim of the acetabulum. The assessed patients lead a normal life, work, often doing a job that is inappropriate for people who should take special care of their hip joints. PMID- 11057011 TI - [Mechanical durability characteristics of polyethylene cup in total hip arthroplasty]. AB - The paper presents an analysis of the processes that determine the durability of the polyethylene cup of the Weller total hip prosthesis. Mechanisms of wear of the polyethylene cup have been presented. It has been inferred that the process of loosening of the polyethylene cup is a combination of plastic deformation, tribologic wear, fracture and changes of the polyethylene structure. All these occurrences are connected with reaction of bone tissue to stress and polyethylene debris. PMID- 11057012 TI - [Heterotopic ossification as a complication of total hip replacement]. AB - Heterotopic ossification is the most common complication after THR. The authors present the distribution of frequency of ectopic ossification after cementless THR using Mittelmeier and Parhofer-Monch prosthesis and it's influence on THR outcome. 151 hip joints were evaluated with a follow-up period of more than 2 years. All procedures were performed by a lateral approach. The ectopic ossification was verified according to the Brooker classification. Clinical evaluation was performed according to the d'Aubigne-Postel method in Charnley's modification. The 3rd degree ectopic ossification was found in 14 hip joint and 4th degree ectopic ossification in 2 hip joints (together 10.6%). The positive correlation between the degree of ossification (3rd and 4th) and the decrease of range of motion of the hip joint along with an increase in pain after THR was found. PMID- 11057013 TI - [Treatment of trochanteric fractures of the femur in elderly patients using Ender rods]. AB - The paper presents the outcome of treatment of patients older than years who sustained trochanteric fractures. Thirty-four females and ten males underwent surgical procedure according to Ender's method. Average age among females was 81 years and among male 72 years. In 63% of the cases a good or excellent functional result was noted. No deaths were noted, neither during hospitalization nor during follow-up. Poor fracture healing nor malunions were noted. Only few local and general complications were observed. The cost of Ender rods is much lower in comparison with DHS, Spiral-Blade or intramedullary implants. PMID- 11057014 TI - [The evaluation of Ender's method for the treatment of trochanteric fractures in patients over 65 with a nail shape change proposal]. AB - During the period between 1991-1998 534 patients aged over 65 years with inter and subtrochanteric fractures were treated with Ender's method in the Bielanski Hospital. The authors' own clinical criteria were used to assess the results. In 80% of the cases the results were either good or satisfactory. The authors focused on the most frequently occurring technical errors during surgery and complications when using this method. The authors present a technique used to bend the nails, which prevents their migration--the most common complication noted when using this method. PMID- 11057015 TI - [Problems, obstacles and complications of femoral lengthening with the use of the Italian modification of the Ilizarov device]. AB - The results of femoral lengthening using the Italian modification of the Ilizarov are presented. Mean age of the patients was 14 years (ranging from 7 to 29). The most frequent etiology of limb shortening was femoral hypoplasia (7 patients) and sequelae of septic arthritis of the hip and/or the knee (6 patients). Indications for surgical treatment were limb shortening from 3 to 12 cm (mean 6.5 cm), along with axial deviation ranging from 10 degrees to 40 degrees in 6 patients. Mean follow-up time was 15 months (ranging from 6 to 35 months). The Ilizarov apparatus was based on two distal rings, stabilized by "K" wires, and proximally by a ring connected with an Italian femoral arch, stabilized by a Schanz screw. Planned lengthening (ranging from 3.5 cm to 12 cm) was achieved in all treated patients. The healing index ranged from 0.8 to 2.1 month/cm (mean 1.4). Problems, obstacles and complications were analyzed according to the Paley classification. In all 16 patients without primary knee stabilization, limited knee flexion ranging from 5 degrees to 90 degrees (mean 40 degrees) was noted during the distraction phase, which didn't improve significantly during the consolidation phase. Knee flexion improved to a mean 90 degrees after a 6 month follow-up. Bone regenerate defects (cysts, narrowing) were noted in 4 patients. Secondary knee stabilization was performed in 2 cases. In the first case because of knee pain and a severe limitation of knee motion. In the second, during a revision procedure because of distal femur angulation. Premature consolidation was noted in one patient and was treated by osteotomy. In one case axial deviation during the consolidation phase required osteotomy. In another case a fracture of the femur was treated by a plaster cast. In one case 1.5 years after the lengthening procedure subluxation of the hip was noted. Permanent knee flexion limitation to less than 90 degrees was noted in 6 patients. Femoral lengthening with the use of the Italian modification of the Ilizarov device give a high incidence of knee range of motion limitation, which can be decreased by preserving more than 30 degrees knee flexion during the distraction phase. PMID- 11057016 TI - [Rehabilitation principles following treatment with the Ilizarov method of the lower extremities]. AB - The paper presents the rehabilitation principles of patients treated by the Ilizarov method. Based on different disease etiology, site and age of the patient, different rehabilitation algorithms are presented. A way of management and rehabilitation of patients treated with the Ilizarov method worked out at the Department of Pediatric Orthopedics at of the K. Marcinkowski University of Medical Sciences of Poznan is also described. PMID- 11057017 TI - [Current methods of thromboprophylaxis in total hip arthroplasty]. AB - The paper presents a review of methods of thromboprophylaxis, with regard given to both pharmacological and physiotherapeutic means in patients who underwent a total hip replacement procedure. Routine procedure should involve proper assertion of indications for surgical treatment, appropriate preoperative planning, surgery carried out in conduction anaesthesia, in the shortest possible time and as atraumatically as possible. Several weeks' of pharmacological treatment involving use of low molecular weight heparins in unison with mechanical means e.g. elastic stockings should follow surgical intervention. Quick mobilization of the patient following the surgical procedure along with intensive physiotherapy results in a decrease of the risk of thrombosis. In order to increase the efficiency of thromboprophylaxis different preventive methods should be applied simultaneously. PMID- 11057018 TI - [MRNA activity in synovial fluid after anterior cruciate ligament reconstruction]. AB - The authors present the results of studies on the transcription level of mRNA (poly A + mRNA) in patients in whom anterior cruciate ligament reconstruction was performed. In a group of 9 patients who underwent this procedure an average increase of the concentration of mRNA was noted--from 1.5 micrograms/ml to 67 micrograms/ml in the first week following surgery. This was followed by a decrease of concentration in the third week to 37 micrograms/ml and 7.8 micrograms/ml in the fifth week. In the control group (16 patients who underwent arthroscopy with partial meniscectomy) an average increase in mRNA concentration from 1.3 micrograms/ml to 4.8 micrograms/ml in the first week after the procedure was noted. It was followed by a decrease to 1.6 micrograms/ml in the third week after surgery. The authors believe the marked increase of mRNA is a result of stimulation of synovial cells and an increase of the number of mononuclear cells in the synovial fluid. The small concentration increase of mRNA in the control group seems to be related only the increase in number of mononuclear cells in synovial fluid. PMID- 11057019 TI - [Prolonged interfragmentary variable compression in the treatment of aseptic nonunion of tibial fractures]. AB - The authors present their own experiences with the treatment of nonunion of tibial fractures through application of prolonged interfragmentary variable compression. This method was applied by the authors in thirty six cases of tibial fracture aseptic nonunion. In 33 cases union with a good functional result was achieved in 17 weeks on average. In 2 cases satisfactory union with minimal angulation of the bone fragments was achieved. In one case the method was found to be unsuccessful. Prolonged interfragmentary variable compression is simple, safe and effective in the treatment of aseptic nonunion of tibial fractures. PMID- 11057020 TI - [Post-traumatic lymphatic and venous drainage changes in persistent edema of lower extremities]. AB - Mechanical injury of soft tissues and bones of the lower extremity is followed by chronic edema at the site of trauma and distally to it. This complication affects almost every patient with a fracture of the lower limb. The question is whether posttraumatic edema is due to lymphatic obstruction, venous thrombosis or both, or a local cytokine and growth factor hyperactivity at the fracture site. The aim of study was to assess the venous and lymph outflow in patient with chronic postraumatic edema of the lower limbs. A group of 19 patients with chronic edema lasting for more than 3 months was evaluated. Limb circumference, tissue tone measurements, skin temperature and Doppler enhanced ultrasonography were all taken down for the 19 patients in the evaluated group. Limb circumference was measured at the following level: foot, ankle, calf and thigh. Results showed an increase of circumference in comparison with the healthy extremity at each evaluated level of: 1.20 +/- 1.65 cm, 1.63 +/- 1.41 cm, 1.40 +/- 1.72 cm and 0.30 +/- 1.90 cm. Local temperature increase compared to the healthy extremity was also noted (0.93 +/- 0.81 degree C and 0.37 +/- 0.21 degree C measured at ankle and calf level). Tissue tone measurements and tone index (a quotient of tone measurement values in the extremity with edema and in the healthy extremity) were also increased by 0.86 +/- 0.57, 0.85 +/- 0.34 and 0.86 +/- 0.28, when measured with 40 g, 110 g and 180 g weights respectively. In 17 cases (89.5%) lymphoscintigraphy demonstrated an increased lymphatic outflow compared to the contralateral extremity. A marked increase in the inguinal lymph nodes was also noted. In the remaining 2 cases (10.5%) extravasation of the contrast medium into the skin indicated lymph outflow disorders. Only in 5 cases (26.3%) ultrasonography indicated deep vein thrombosis. The obtained results indicate that the pathophysiology of chronic postraumatic edema is linked with an inflammatory and restorative reaction at the fracture site. Only in a limited number of cases deep vein thrombosis and damaged lymphatic vessels are responsible for postraumatic edema. PMID- 11057021 TI - [Molecular basis of achondroplasia, hypochondroplasia, and thanatophoric dysplasia]. AB - Fibroblast growth factor 2 (FGF2) inhibits proliferation and hypertrophy of chondrocytes in the growth plate, synthesis of cartilage matrix, terminal differentiation of hypertrophic chondrocytes and matrix calcification. Recent studies have found that mutations in the receptor for fibroblast growth factor 3 (FGFR3) cause achondroplasia, hypochondroplasia and thanatophoric dysplasia. These mutations evoke uncontrolled stimulation of the receptor, leading to inhibition of bone growth. Inactivation of the receptor in experimental animals causes excessive chondrocyte proliferation and abnormal bone length. Chondrocyte stem cells proliferate in the ossification groove of Ranvier and contribute to both peripheral and longitudinal growth of the growth plate. They express FGFR3, have a potential to differentiate into chondrocytes and are therefore considered adequate for healing cartilage defects in the articular surface. It is at present unknown what happens to the chondrocyte precursor cells in the ossification groove of patients with FGFR3 mutation. PMID- 11057022 TI - [Provocation tests in clinical examination of carpal instabilities: part I]. AB - Basing on a review of available literature, the author presents 9 provocative tests used in diagnosing carpal instability. Their methodology and principles of clinical interpretation are reviewed. Test used to diagnose instability in the following joints: radio-carpal, scapho-lunate, luno-triquetral and carpo metacarpal II-V are discussed. PMID- 11057023 TI - [Effect of water storage and intrapulpal pressure on microleakage of three restorative materials]. AB - Three different restorative materials, Z100 composite, F2000 compomer and Vitremer glass ionomer cement are currently proposed for Class V restorations. The aim of this in vitro study was to evaluate the influence of water storage and the simulated intrapulpal pressure (sIP) on the quality of the margins of class V restorations located both in enamel and dentin. The water resorption of restorative materials containing hydrophilic groups (compomers and glass ionomer cements) can favourably modify the marginal sealing ability by hydroscopic expansion. The influence of the sIP was specific to the material. While F2000 compomer and Vitremer glass ionomer cement were un-influenced by sIP, with Z100 composite a significant difference could be observed. It was concluded that F2000 compomer and Vitremer glass ionomer cement showed significantly less microleakage, which means a better marginal sealing ability than Z100 composite. PMID- 11057024 TI - [Some depth-psychological aspects of the psychogenic symptoms of oro-facial tissues]. AB - The specificity of the oral psychogenic manifestations is based on the unique psychological and depth-psychological function of the mouth, and teeth during infancy (breasting), sexuality, and aggressivity. Fixed dentures, implants, and especially removable dentures can modify strongly these psychological functions and activate oral fixations leading to psychogenic symptoms. In the present paper the great importance of the oral phase of the development of personality and the symbolical values of the mouth, teeth, tongue, and face in the oro-facial conversive symptom manifestations are discussed. PMID- 11057025 TI - [Morphologic alterations of tooth crown in patients with Turner syndrome and its association with orthodontic anomalies]. AB - In females with numerical and/or structural aberration of the X chromosome as in Turner's syndrome, somatic development deviates from the normal pattern. The aim of the present study was to investigate the tooth crown dimensions, and report the size of the alveolar arch of the maxilla and the mandible of patients with Turner's syndrome. The study population consisted of 29 girls with Turner's syndrome and 30 healthy female controls. All mesiodistal dimensions were significantly smaller in Turner's syndrome compared with those of normal controls. The labiolingual dimensions were smaller in the patient group with the exception of the canines and the lower central incisors. The alveolar arch of the maxilla was narrower and of normal length, but the mandibular arch was shorter and broader. Orthodontic anomalies were found more frequent and serious of patients with Turner's syndrome. Our examination showed the importance of early diagnosis of oral anomalies as well as dental treatment of patients with Turner's syndrome. PMID- 11057026 TI - [Care of patients with oral cavity tumors in the hospital and at home]. AB - This study reports on experience acquired during the care of patients who could not participate in surgery, chemotherapy or radiation therapy because of the extent of the primary, recurrent or metastatic tumour, or because of the deterioration of the general condition or (in the event of recurrence), intervention with a curative aim could not be performed for the mentioned reasons. In these cases, improvement or preservation of the quality of life is the fundamental goal to be achieved at all times, for these patients must receive the best possible supportive treatment for the remainder of their lives, Unfortunately, the quaranteeing of this often tends to become mainly a financial question rather than a professional one. The medical staff has very little chance to influence the financial aspects, and must therefore do everything possible to ensure the highest possible level of care during hospitalization. Attention is drawn to the significant roles to be played by the family, the family doctor, the various home-care services and the hospice. PMID- 11057027 TI - Developmental changes of MAP2 immunoreactivity in the hippocampus proper and dentate gyrus of the rat. AB - Microtubules are present in high concentration in the nervous system and are a prominent component of the neuronal cytoskeleton. Microtubules are composed of tubulin and variety of microtubule-associated proteins (MAPs), which have been implicated in the regulation of microtubule assembly and function. MAP2 is the most abundant of these proteins, and it has been extensively characterized in various functional and pathological conditions. In the present study the distribution of MAP2 was examined in each layer of the CA1 and CA3 regions of the hippocampus proper and dentate gyrus in rat development. A total of 40 brains at various ages starting from postnatal day (P) 0 to P90 were examined. After perfusional fixation the brains were frozen and cut on the coronal plane and stained with either cresyl violet or standard immunohistochemical methods using the anti-MAP2 antibody. MAP2 exhibited a somatodendritic pattern of localization in cells of the hippocampus. Staining was most prominent in dendrites and perikarya as well as granules surrounding cell bodies. In a newborn rat's brain immunostaining was intense in granules and faint in perikarya. Between P4 and P21 immunostaining density for MAP2 was stronger and appeared in perikarya, granules, and dendritic trees. After P21 the perikarya and dendrites of the pyramidal layer and stratum radiatum of the hippocampus proper, as well as the molecular and granular layer of dentate gyrus, showed reduced immunoreactivity. In the stratum oriens of the hippocampus and polymorphic layer of the dentate gyrus immunoreactivity was still strong until P90. PMID- 11057028 TI - Dystrophinopathies in females. AB - Various laboratory tests were performed to establish carriership in 24 familial and sporadic carriers of Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD). The activity of creatine kinase was in all females but one, very high and significantly higher in isolated carriers; quantitative EMG indicated myopathic changes, muscle biopsies revealed different degrees of changes--from a variability of muscle fibers size and central nuclei to severe dystrophic features. Immunohistochemical evaluation of dystrophin revealed, in all females but one, mosaic pattern of staining--a mixture of dystrophin-positive and dystrophin-negative fibers, the latter consist 15-30% of all fibers. Quantitative evaluation of dystrophin showed a reduced abundance with normal or abnormal molecular weight. The abnormalities were more expressed in sporadic cases. The detection of sporadic carriers, particularly the non-manifesting clinical, is a very important progress--it permits the correct diagnosis (before, these females were diagnosed as limb girdle muscle dystrophy (LGMD) and supply them with the benefit of genetic counselling, which also requires some modification. PMID- 11057029 TI - Extracellular signal-regulated kinase suppression is not involved in apoptosis of neuroblastoma N2a cells induced by protein kinase C inhibition. AB - A model study for the mechanism of cell death in vitro was established on the neuroblastoma N2a cell line. By differential staining of living cells with Hoechst 33258 and propidium iodide, or by Terminal-dUTP-Transferase-Nick-End Labelling (TUNEL), cJun/AP1 immunoreactivity, and DNA laddering, we show that the N2a cell line responded with apoptosis to protein kinase C (PKC) inhibition. Two different classes of PKC inhibitors (staurosporine and Go6976) at concentrations generally regarded as PKC-selective (10 nM and 50 nM, respectively), significantly increased the number of apoptotic cells in N2a cultures. The cells started to die 2-3 hours after the treatment; then, at 6 and/or 24 hours, approximately 30-40% of the cells acquired the apoptotic feature. This response was dependent on neither cell differentiation nor PKC and bcl2 gene and protein expression, but exclusively on the concomitant withdrawal of serum from growth medium. Furthermore, both of the experimental manipulations (serum deprivation and PKC inhibition) synergically, but to different extents, suppressed the extracellular signal regulated kinase (ERK) pathway. Their pro-apoptotic effect, however, was neither mimicked nor modified by an additional inhibition of MEK/ERK kinase by 50 microM PD 98059, resulting in an 80% inhibition of its initial activity. We conclude, therefore, that apoptosis of N2a cells triggered by PKC inhibition, as well as its abrogation by serum, is totally independent of concomitant modulations of ERK activity also evoked by the above treatments. PMID- 11057030 TI - Epithelioid schwannomas of the acoustic nerve. AB - Epithelioid schwannomas occur predominantly in relation to peripheral nerves and are associated with histological and clinical malignancy. However, a variant of the epithelioid schwannoma involving cranial nerves is extremely rare. In this study we report three cases of epithelioid schwannomas originating from the acoustic nerves and located in the cerebello-pontine angles. In the first case, the tumor was histopathologically entirely solid and demonstrated biphasic pattern with both spindle-shaped cells and a population of round or polygonal epithelioid cells. The second one consisted of the smaller part exhibiting typical Antoni B and A tissue and large areas containing clusters and bundles of epithelioid cells. Purely epithelioid schwannoma composed predominantly of cords or nests of round and polygonal epithelioid cells were observed in the third case. All schwannomas revealed marked polymorphism and nuclear hyperchromasia. Immunohistochemical studies showed a diffuse, strong positivity for S-100 protein in the cytoplasm of the spindle and epithelioid tumor cells. These two populations of cells were positively stained for vimentin, but were negative for EMA, cytokeratin and HMB45. Patchy GFAP-immunoreactivity was also noticed at the peripheral parts of the tumors. The authors discuss differential diagnosis of this unusual variant of schwannoma in relation to malignant transformation of the epithelioid component. PMID- 11057031 TI - Paraneoplastic syndrome in the course of lung adenocarcinoma: morphological picture and immunohistochemical analysis of the inflammatory infiltrates and PECAM-1 expression. AB - We examined sections of brain, spinal cord, spinal roots, and peripheral nerves from a patient with paraneoplastic syndrome in the course of lung adenocarcinoma. Morphological examination showed marked loss of myelin fibers in peripheral nerves, severe brain tissue edema, and paraneoplastic degeneration involving cerebrum and cerebellum with inflammatory components. Inflammatory infiltrates examined immunohistochemically using antibodies against antigens CD 3, CD 4, CD 8, and CD 20 turned out to be composed of cytotoxic T lymphocytes. The expression of platelet-endothelial cell adhesion molecule-1 (PECAM-1) in blood vessels was increased in comparison with control material, which may facilitate transendothelial lymphocyte migration triggering a cascade of biochemical and morphological reactions observed in paraneoplastic syndrome. PMID- 11057032 TI - The effect of moderate hypoxia on free sterols: content and pattern in white matter. AB - Wistar rats underwent moderate hypoxia by exposing them to a respiratory gas mixture containing 7% oxygen, for 30 min. The myelin was analyzed at 4 h 24 h, day 14, and day 60 after hypoxia. The group of neutral lipids isolated from myelin was analyzed by GC-MS (gas chromatography-mass spectrometry) to separate and measure free sterols. In the myelin of hypoxia treated animals the pattern of free sterols was variable. Cholesterol concentration decreased with the changes in the content and composition of sterols present in smaller amounts. These changes may indicate the disturbances in some steps of cholesterol biosynthesis and metabolism in pathologic conditions. The appearance of sterols characteristic for plants (beta-sitosterol) was noticed in myelin of experimental animals. The possible reason for the presence of this sterol in animals may be the result of its intake during feeding, however, biosynthesis can not be excluded. PMID- 11057033 TI - Determining the feasibility of diagnosing meningiomas using static teleneuropathy images transmitted electronically. AB - The feasibility of using still images sent via the Internet for remote neuropathological consultation was examined. We assessed the diagnostic agreement between three groups of neuropathologists and a reference laboratory. All groups independently evaluated pictorial data exchanged by e-mail or ftp server. In the group of benign meningothelial meningioma agreement was reached in 100%, 100%, 92% of cases. In three cases of malignant meningioma agreement was achieved in 100%, 33%, 33% of cases, and in ten cases of atypical meningioma agreement was achieved in 50%, 30%, 40% of cases. Average concordance for all three groups was 83.33%, 54.33%, 57.67%. Our experiment showed that there were no discrepancies in the typical cases of benign meningothelial meningiomas. Disagreement related to the grade II and III lesions i.e. atypical and malignant meningioma. Atypical as well as malignant meningioma seems to need more direct discussion to achieve consensus. The main problem was the subjectivity of the local pathologist who overstated the informational content of the images, and an insufficient number of pictures. This may be prevented in the future by employment of dynamic teleneuropathology using a very high-resolution camera. PMID- 11057034 TI - Real-time teleneuropathology for a second opinion of neurooncological cases. AB - A teleneuropathology system directed by Java programs through a standard Internet browser was evaluated in the present study. Assessment of neurooncological cases was done by remote microscope through the "Case Study" Web page at the Department of Pathology Web site (http:?ampat.amu.edu.pl). The site was used to control a remote automatic microscope, the Axioplan 2 (Zeiss), which was connected to a computer that acts as an Internet server. The Java program for the control of the microscope server is automatically downloaded and started if the user selects the Web site of the corresponding microscope server. The microscope server receives microscope operation commands from the telemicroscopy clients, executes them, and distributes the new microscope image to all of the connected telemicroscopy clients. Fifteen cases were evaluated over several weeks. The percentage of correctly classified cases sent by remote consultation was 100%. Since the system does not require specialized software for the remote side and since the number of possible discussion partners is unlimited, this system may help to overcome obstacles to the practice of teleneuropathology. PMID- 11057035 TI - Aluminum enhances glutamate-mediated neurotoxicity in organotypic cultures of rat hippocampus. AB - Both, glutamate (GLU) and aluminum (Al) have been implicated in neuronal damage and/or death in certain human neurodegenerative disorders. Recent evidence suggests that aluminum (Al) may potentiate the increase in glutamate-induced intracellular calcium overload. The present ultrastructural study was undertaken to determine the effect of Al on the development of GLU-mediated neurotoxicity in tissue culture conditions. The experiments were performed on organotypic cultures of rat hippocampus treated with low, subtoxic concentration of GLU (50 microM) and AlCl3 (400 microM) added to the growth medium separately or in combination. The exposure of cultures to GLU in the presence of Al3+ ions for up to 24 hours resulted in the development of typical excitotoxic neuronal changes, whereas separate GLU treatment at subtoxic doses or single Al application did not produce any apparent tissue damage. The neuronal lesions resulting from the combined application of GLU plus Al consisted predominately of more or less pronounced mitochondrial abnormalities, which are characteristic for early excitotoxic events. Severe swelling of the mitochondria led to the disruption of their internal structure and finally resulted in an apparent microvacuolization of the perikaryal cytoplasm of some pyramidal neurons. The present morphological data evidenced that Al is capable to potentiate the GLU-induced degenerative changes in hippocampal neurons in vitro. This supports the view of a possible role of Al in the process of neurodegeneration and suggests that Al may participate in the development of glutamate-mediated excitotoxic neuronal injury under certain pathological conditions. PMID- 11057036 TI - The influence of the intraventricular hemorrhage on the intracranial pressure and hemodynamic changes in the experimental intracerebral hemorrhage in the rabbits. AB - The experimental model of the intracerebral hematoma in the rabbit was used for the investigation of the changes of the intracranial pressure and selected hemodynamic parameters. The study was performed on 13 adult rabbits, divided into two groups receiving 1 ml (group I--6 animals) and 2 ml (group II--7 animals) of fresh arterial blood, respectively. The monitoring of the intracranial pressure (ICP), the mean arterial blood pressure (MABP), the heart rate (HR), the end tidal CO2 concentration (ETCO2) and the body temperature was measured every minute in the hematoma production phase and every 5 minutes for the consecutive three hours. The volume of the hematoma was calculated according to Cavalieri formula, with the use of the system for the automatic picture analysis. The mean volume of the intraparenchymal part of the hematoma in group I was higher than in the group II. However, in all the representatives of the second group the evidence of the intraventricular hemorrhage was present. The dynamics of the ICP, MABP and HR changes differed significantly in both groups during the period of the observation. On the basis of the physiological and morphological observations we conclude that the changes of ICP remain the most sensitive and valuable parameter during the early course of the intracerebral hemorrhage. Coexistence of the rapid ICP, MABP and HR changes must be always regarded as the possible sign of the intraventricular hemorrhage. PMID- 11057037 TI - Astrogliosis and blood vessel development during human spinal cord myelination. AB - The aim of our study was to investigate a relationship between vessel development, astrogliosis and myelination in humans. We examined spinal cords of human fetuses and infants using immunohistochemical methods with antibodies against Myelin Basic Protein (MBP), Glial Fibrillary Acidic Protein (GFAP) and lectin Ulex Europaeus Type 1 (UEA-1). Our investigation showed that in parallel to the increase of MBP reactivity in the spinal cord white matter more GFAP immunoreactive astrocytes appeared. Reaction of vessel endothelium with lectin UEA-1 revealed the temporal increase of vascularization in the course of the spinal cord tracts myelination in fetuses. In the postnatal period when the myelination is nearly morphologically completed, the spinal cord white matter was poorly vascularized although GFAP-immunoreactive cells were still relatively numerous. We suggested that the increase of vascularization and astrogliosis observed during spinal cord tracts myelination may be connected with their participation in process of myelin sheath formation via, among others, local secretion of growth factors necessary for normal myelin development. PMID- 11057038 TI - Prosencephalic commissures dysgenesis combined with spinal neural tube defects. AB - The neuropathological picture of four cases of prosencephalic malformations coexisting with spinal dysraphic anomalies inclined us to discuss the nosological position of these abnormalities. The hypoplastic anomalies of prosencephalic commissures combined with well-cleaved (except for basal ganglia) brain hemispheres allow to think of failure within lamina terminalis. The observations support the opinion that described malformations are consequence of abnormal planning of prosencephalic formation. Our description testifies ones more that as a result of prolonged time of injury a complex picture of developmental anomalies is noted. PMID- 11057039 TI - Major histocompatibility complex class II expression in the frontal and temporal lobes in the human fetus during development. AB - Antigen-presenting cells (APCs) that show the expression of major histocompatibility complex (MHC) class II molecules in adult persons are related to an early phase of immunological response. These molecules are responsible for the binding, transport, and presentation of a foreign antigen to helper T lymphocytes and determine the type of antibodies produced. They also stimulate the multiplication of specific B lymphocytes and participate in the elimination of autoreactive lymphocytes and maturation of T lymphocytes. Cells with MHC II molecules expressed on their surface were observed in the frontal and temporal lobes of 30 brains of human fetuses with normal development between gestation weeks (GW) 11 and 22. MHC II expression was noted during the whole interval under study. Its immunocytochemical localization was noted on the surface of the cerebral meninges cells, in the choroid plexus of the lateral ventricle and blood vessel lumen, and in ameboid microglia (AM) and ramified microglia (RM) cells in both cerebral lobes of the human fetus. The expression of MHC II that occurred on the cells of the central nervous system (CNS) already in GW 11 may be evidence not only of an early capability of immunological protection of the fetal nervous system, but also of a significant role potentially played by this system in normal embryogenesis. Despite continuous controversy over the cellular lineage of microglia origin, the expression of MHC II on AM and RM cells indicates its mesodermal origin, as in other APCs. PMID- 11057040 TI - Telecommunication--a new tool for quality assurance and control in diagnostic pathology. AB - Telepathology is the diagnostic work of a pathologist at a distance. It includes specific application fields which require specific system solutions. These comprise: a. frozen section service; b. expert consultations; c. remote control measurements, and d. education and training. Applications but, in addition, all aspects of diagnosis, especially those involved in the daily work flow of a pathologist. The image quality, transfer rates, and screen resolution of telepathology systems are sufficient for an additional or primary judgment of histological slides and cytological smears. It is, therefore, possible to include this technique into intralaboratory quality control of all steps of diagnostic procedures and to use this technique for quality assurance and control. Diagnostic quality is not a well defined term, and depends, in addition to external circumstances, on the diagnosis itself. The diagnostic quality can be maintained or even improved by use of telepathology systems. They can serve for control of image quality, access to various information sources, simultaneous transfer of images and diagnosis, and continuous education of the involved pathologists. For example, in continuous education and training, as a first step the pathologist should have access to an image data bank storing images related to the specific case, for example images taken from the same organ and from patients of similar age. The final stage would be an intra-diagnostic learning from the specific case; i.e. the time difference between questionnaires for a difficult diagnosis and the final diagnosis will become a minimum. Automated storing of the specific and diagnosis-associated images of the case will permit an open diagnostic system which will itself adjust to the micro-environment of the department of pathology. Although this future scenario has not been implemented to our knowledge, there is little doubt the development in telepathology will promote the technical procedures in the environment of a diagnostic pathologist. Telepathology is, therefore, not a substitute of conventional diagnostic procedures but a real improvement in the world of pathology. PMID- 11057041 TI - Technological requirements of teleneuropathological systems. AB - Teleneuropathology is the practice of conducting remote neuropathological examinations with the use of telecommunication links. Because of a limited number of expert neuropathologists, some, especially smaller departments have the equipment to conduct the examination but do not have a specialist who would be able to evaluate material from the central nervous system. In case of teleneuropathology, a neuropathologist examines tissue fragments taken during an operation by means of a telemicroscope connected with the computer through a telecommunications network. It enables the neuropathologist to operate the microscope and camera remotely. Two basic systems exist for performing remote neuropathological examination: static and dynamic. Both have different needs in medical, computing and telecommunication aspect. Depending on the type of service the public telephone network, the integrated services digital network, or optical fibre should be used. Conditionally Internet can be used as a link for teleneuropathological system. However, for the newest developments in teleneuropathology such as teleconference and remote operation on robotized microscope only transmission over the integrated service digital network, which guarantees high speed of transmission gives a possibility to communicate. Because images are basic information element in teleneuropathological systems the high capacity of acquisition, processing, storing, transmission, and visualization equipment is necessary. The farther development of telecommunication as well as standardization of recording and transmission procedures of pictorial data is necessary. PMID- 11057042 TI - [Population genetics of Chinese surnames. I. Surname frequency distribution and genetic diversity in Chinese]. AB - This paper deals with the distributional characteristics of Chinese surnames and the factors affecting their distribution. Analysis of alpha and nu values for three periods in the last 1,000 years clearly reveals the distributional pattern of Chinese surnames. In China, communities with the same surname are widespread, especially in the central areas where there are more relatively isolated Y chromosome of the same source (communities with the same surname) and more surnames than in other areas (especially the South China). If each surname has a certain kind of mono-type Y-DNA, then, according to the distribution of surnames, there exist the most mono-type of Y-DNA in the central provinces where are the originating center of Chinese surnames. For the first time, an equation is constructed predicting the kind of Chinese surnames. It has been estimated that there are about 3,100 surnames present in use for the Han nationality. PMID- 11057043 TI - [Genescan for STR analysis and genetic distribution in a population sample from Han, China]. AB - Genetic distributions for nine STR loci and Amelogenin locus were determined in a Chinese Han population based on DNA sequencing. The databanks in Chinese Population were generated by using Genescan, genotype and genetic distribution analysis. Allele frequency distribution was determined for 10 loci, such as D3S1358, VWA, FGA, Amelogenin, THO1, TPOX, CSF1PO, D5S818, D13S317 and D7S820. The results show significant differences between ethnic groups (African-American, US-Caucasian and Chinese Han) in the pattern of distribution as well as in the related frequency of the most common alleles of their STR loci. The probabilities of identity values for the population described in this section are 2.79 x 10( 10) (U.S-Caucasian), 1.23 x 10(-10) (African-American) and 0.5 x 10(-10) (Chinese Han). The results show the probability of Paternity Exclusion (PPE) values of 0.9998 (Chinese Han), 0.9996 (African-American) and 0.9994 (U.S-Caucasian). These results suggested that the nine STR loci and the Amelogenin locus are very useful for human identification, such as analyzing forensis casework, establishing DNA databanks, processing paternity test, evaluation linkage genetics, studying gene natural resources and monitoring bone marrow transplants. PMID- 11057044 TI - [Research on constructing phylogenetics trees of ruminants basing on the database of milk protein gene sequences]. AB - Primers designed according to the sequences of four milk protein genes of cow Bos taurus (alpha-lactoalbumin, beta-lactoglobin, beta- and kappa-casein) were used to amplify the full length gene of alpha-lactalbumin in yak Bos grunniens (2999 bp), water buffalo Bubalus arnee bubalis (278 bp), partial sequence of this gene in red deer cervus elaphs xanthopygus (1582 bp), 5' and 3' flanking region of beta-lactoglobin gene (2167 bp and 1096 bp in length respectively), 5'-flanking region and exon VIII to exon IX of beta-casein gene (987 bp and 1096 bp in length respectively), exonIV of kappa-casein gene (780 bp). All the amplified DNA fragments were cloned and the Nt sequences were determined. Phylogenetic tree containing 20 species (or subspecies) of ruminantia suborder was constructed according to the partial sequence of kappa-casein gene exon IV (363 bp in length), which shows good monophyly of the Bovidae. And trees constructed according to other milk protein genes indicate that all the milk protein genes have good features for drawing phylogenetics tree at least among species belonging to different subfamilies. PMID- 11057045 TI - [Studies of the genetic polymorphism of Schistosoma japonicum (Chinese mainland strains)]. AB - In order to analyze the interspecies diversity and the extent of diversity among Schistosoma japonicum Chinese mainland strains, the genetic variation on gene level among 6 isolates collected from Jiangxi, Hunan, Hubei, Anhui, Sichuan, Yunnan and a cultured isolate from a laboratory were studied using molecular biological technique. There were only two different bases at position 112 and 143 in 536 bp sequence of 28S rDNA-D2 domain between Anhui and Yunnan isolates, the homology was 99.6%. The result could be explained the reason of why the migration rates of single chain 28S rDNA-D2 domain among the above 7 isolates were the same completely in PCR-SSCP. However, comparing with the sequences of the Philippine isolate of S. japonicum, S. mansoni and S. aematobium, there were 6, 94 and 93 different bases and the homologies were 98.9, 82.5 and 82.7 per cent separately. With 8 restriction endonucleases to analyze the ITS of rDNA obtained by PCR from the 7 isolates, the results showed that only 3 minor bands were different, e.g. 5.3% of total 58 fragments. It was suggested that the ITS of rDNA among 7 isolates were highly conserved. Using randomly amplified polymorphic DNA (RAPD) to analyze the genetic diversity of the genomes of the 7 isolates, the average genetic distance (D) calculated from total 284 amplified fragments was 0.22. The maximum D was 0.30 and existed between Anhui and Yunnan isolates. The minimum D was 0.13 and existed between Sichuan and Yunnan isolates. The clustering analysis of genetic distances showed that the 7 isolates could be gathered in one group. From above three results, it could be considered that the genetic diversity on gene level among S. japonicum Chinese mainland strains was very low. PMID- 11057046 TI - [Studies of development of disomic addition lines of Triticum aestivum-Haynaldia villosa via AABBDDDD octaploid]. AB - Hexaploid hybrid between Triticum aestivum-Aegilops squarrosa (2n = 8x = 56, AABBDDDD) octaploid and Triticum durum-Haynaldia villosa (2n = 6x = 42, AABBVV) was self pollinated. The putitative T. aestivum-H. villosa 6V and 2V addition lines were creened by C-banding in F4 and chromosome configurations of PMC at MI were 0.14I + 20.42II + 1.50II and 0.10 I + 20.07II + 1.82II, respectively. Genomic DNA of 95-7 and 26-7 were digested by EcoRI, and Southern bloting was employed using group 6 probe Psr113 for 95-7 and group 2 probe BCD240 fro 26-7. The results showed that 95-7 and 26-7 had the same special bands as H. villosa did. Therefore, 95-7 and 26-7 further identified were T. aestivum-H. villosa 6V and 2V disomic addition lines. PMID- 11057047 TI - [High efficient intergeneric chromosomal translocations between wheat (Triticum aestivum L.) and Dasypyrum villosum arising from tissue culture and irradiation]. AB - Intergeneric chromosomal translocations were discernable both in callus cells and in regenerants arising from crosses between Triticum aestivum and T. durum Dasypyrum villosum amphiploid c.v. TH1 and TH1W by means of fluorescence in situ hybridization. There were not only reciprocal translocations, but small fragment translocations. The results proved again the feasibility of creating intergeneric translocations via tissue culture. Irradiation facilitated numerical and structural chromosome changes in callus cells. The frequency of translocations was as high as 7.4 percent in irradiated callus cells. Callus age had an important impact on numerical and structural chromosome abnormalities. During a given time of culture, the frequency of unchanged cells was declined, while those cells with chromosome losses were inclined. The duration of culture had not significant effects on cells with chromosome gains. As structural chromosome changes is concerned, the duration of culture predominantly increased the frequency of cells with telocentric chromosomes. The chromosomal changes occurred at the initiation period of tissue culture. A number of cells which were doubled their chromosome numbers (2n = 84) were observed at a certain frequency in the period of tissue culture. These cells, however, disappeared in the successive culture. PMID- 11057048 TI - [The genetic behavior of an Agropyron intermedium chromosome conferring BYDV resistance in wheat background]. AB - Based on the chromosome origin identification of an Agropyron intermedium chromosome with BYDV resistance in Yi4212, a BYDV-resistant substitution line derived from the hybrids between 77-5433 and Zhong5, its ability compensating the lost wheat chromosomes and its transmission rate were investigated. The results indicated that the BYDV-resistant chromosomes originated from Zhong 5 could compensate the wheat chromosomes of homotologous group 2, 5 and 7. The chromosome conferring resistance to BYDV showed the intendance to substitute 2D rather than 2A, 2B wheat chromosomes in disomic substitution lines. In (77-5433/Zhong 5) selfing F2 population, 10 kinds of chromosome constitutions, including 9 expected and 1 unexpected kinds, and much chromosome number variation and little chromosome structure variation were observed. The transmission rate of the BYDV resistant chromosome and the gametes carrying this chromosome were 56.3% and 33.0% respectively, both are lower than the expected ratio 75.0% and 50.0%. Some reasons were given to explain the related results, and chromosome in situ hybridization was an efficient and accurate way in studying the alien chromosome behavior in wheat background. PMID- 11057049 TI - [Alien chromosomal identification in VE161 wheat]. AB - VE161 wheat which has the function of promotting homoeologous pairing is a male sterile substitution line or a fertile addition line with one pair of alien chromosomes from Agropyron elongatum. The alien chromosome in VE161 wheat was detected by means of techniques of total genomic DNA in situ hybridization, RFLP and double-ditelo chromosomal analysis. The results from in situ hybridization confirmed that the alien chromosome was derived from Agropyron elongatum. The RFLP analysis showed that it belongs to fourth homoeologous group, and the results from double-ditelo chromosomal analysis gave evidence that 4B chromosome was replaced in the substitution line, so it can be concluded that, in VE161 wheat, the alien chromosome would be 4E and the replaced wheat chromosome would be 4B in the substitution line. The reason for the replaced chromosome identified as 7B in a earlier study might be due to the rearrangement occurred in the genome of wheat. PMID- 11057050 TI - [Transmission and genetic effect of ting chromosome in cross progenies from octoploid Tritileymus with 4D nullisomic]. AB - A pair of ting chromosomes (ti) added to 72180 4D nullisomic was subcentral centromere chromosome and was about 1/3-1/4 the size of a chromosome on the average length. It's chromosome configuration was 19.59" (18-20) + 0.46' (0-4) + 0.99" (0-1) + 0.96ti" (0-1) + 0.08ti' (0-2) at PMC MI. The cells with ring ti" account for 96.19%, of which 82.38% were free on each side of the equatorial plane. ti chromosome paired in advance, defered segregation and had no synapsis with a chromosome. When the cross progenies from 72180 4D nullisomic with octoploid Trilileymus were backcrossed to the 4D nullisomic used as the male, the transmitting rate of 2ti chromosome was six times higher than that where the nullisomic was used as the female. The transmitting and loss rates of ti chromosome were 50.38% and 18.34% respectively, and averagely each plant has 0.72 ti chromosome in the cross generations. The appearing rate of ti chromosome was 43.18% in root tip cell and 8.16% in PMC MI of the plants without ti chromosome. When the number of a chromosome was less than 42, the genetic effect of ti chromosome on the quantitative character was significant, while the number of a chromosome was 42, the genetic effect was not apparent. PMID- 11057051 TI - [Genetic diversity and differentiation of cultivated Fagoyrum tataricum populations from three counties in south Liangshan Automomous Prefecture of Yi Nationality, Sichuan, China]. AB - Genetic diversity and differentiation among 8 cultivated populations of Fagopyrum tataricum from the counties of Jinyang, Leibo and Miyi in south Liangshan Autonomous Prefecture of Yi Nationality, Sichuan Province, China were investigated using allozyme electrophoresis. The allozymic diversity is roughly correlated with agrobiological features. The results suggest that genetic diversity of F. tataricum is quite high. The mean number of alleles per locus A is 1.8; the percentage of polymorphic loci P is 46.6%, and the mean observed heterozygosities Ho and the mean expected heterozygosities He are 0.187 and 0.218 respectively, the ratios of gene diversities of heterozygosities Fsr is 0.22, which indicates that there is 22 percent of genetic differentiation among populations, and suggests that for protecting genetic resources all the populations should be included in. PMID- 11057052 TI - [A new model of translational control of gene expression in polycistron++ of AB5 entrerotoxin]. AB - The expression level of the B subunit gene of cholera toxin (ctx) and E. coli heat labile toxin (ltx) is five to seven times more than that of A subunit gene. In these studies, a 80 basepair translation regulation element was found located in the structure gene of A gene of both toxin operon which consists of three translation initiation region. Site-directed mutation of the initiation codon of TIR3 resulted in the 9 time decrease of the expression of the downstream cistron which was translational coupled with A gene. The results indicated that translation from the internal of A gene and translation coupling are responsible for the differential expression level of the A and B gene of AB5 enterotoxin. PMID- 11057053 TI - [FruA, a transcription factor in Myxococus xamthus, regulates transcription of target genes in collaboration with the associated protein FruB]. AB - Many developmental genes are regulated by FruA, a transcription factor essential for the development of Myxococus xamthus. Another protein, designated FruB, was purified from myxobacteria by its affinity to FruA. FruB could be phosphorylated by protein kinase(s) located in cell membrane. Gel shift assay showed that FruA regulates transcription of target genes in collaboration with phosphorylated FruB. This study may shed light on the molecular mechanisms of regulatory network involved in the development of Myxococus xamthus. PMID- 11057054 TI - Individual sociodemographic characteristics associated with hospitalization for pediatric ambulatory care sensitive conditions. AB - This study examined the association of individual sociodemographic characteristics with pediatric ambulatory care sensitive condition (ACSC) hospitalizations in American hospitals while controlling for selected hospital characteristics. Data came from the 1994 National Hospital Discharge Survey. Bivariate statistical comparisons were performed to test the differences between ACSC and non-ACSC hospitalization rates in patient demographic and hospital characteristics. Logistic regression was followed to examine the relative significance of patient and hospital characteristics associated with pediatric ACSC hospitalizations. Such individual sociodemographic characteristics as age, race, and insurance status were significant predictors of ACSC hospitalization. Younger children were more likely to have ACSC hospitalization than older ones. Black children were 1.653 times more likely than white children to be hospitalized for ACSC (confidence interval = 1.53-1.79). Those with Medicaid or no secondary insurance were more likely to be hospitalized for ACSC than those with private insurance or with secondary insurance. PMID- 11057055 TI - Service learning: a strategy for conducting a health needs assessment of the homeless. AB - An agency providing health care services for homeless persons and a nursing department at a liberal arts college established a service-learning partnership to complete a health needs assessment of homeless persons. Under the guidance of agency staff and a nursing faculty member, seven nursing students surveyed shelter residents (n = 101) in four urban shelters and conducted a focus group to identify residents' perceptions of health, health care needs, and health care service delivery. The service-learning partnership expanded the agency's services by providing research consultation and data collection that resulted in recommendations to improve health care services for the homeless. The agency contributed to the education of health professionals by providing students with a meaningful community service experience. PMID- 11057056 TI - Insurance and education determine survival in infantile coarctation of the aorta. AB - Health outcomes are determined by case severity, physician decisions, and patient variables. In a population-based study between 1981 and 1989, 103 cases of infant coarctation of the aorta were diagnosed before one year of age. The goal of this study was to determine whether patient race, gender, income, and insurance status had effects on outcome of coarctation of the aorta that were distinct from the effect of case severity. Survival of infants with coarctation of the aorta, a common congenital cardiovascular malformation, is associated with greater maternal education and with having any health insurance but not with measures of severity. Infants without health insurance are 12.8 times more likely to die than infants with any health insurance. Fifty-five percent of all deaths in infant coarctation occur prior to surgical treatment. One-third of deaths occur without diagnosis. Outcome measures require knowledge of the entire population and of insurance status to inform policy. PMID- 11057057 TI - Caregivers' unmet needs for support in caring for functionally impaired elderly persons: a community sample. AB - Few estimates have been made of the extent to which the needs of caregivers are met. In addition to the inadequate capacity of services, many caregivers lack adequate financial resources, social resources, or other means to access them. Caregivers who provide services to minority or poor elderly may be particularly needy since their care receivers tend to be less healthy and are less likely to use institutional facilities. To address this issue, the authors studied a community sample of 124 caregivers who identified correlates of their perceived unmet caregiver needs and their use of supportive services available for their caregiving. Results indicated that 51.8 percent of women and 67.4 percent of men reported needs for one or more community services that were not met. It was concluded that caregivers who are poor or who required financial assistance are at the highest risk for needing assistance while providing caregiving services. Community services may more effectively target potential needs of caregivers through routine screenings. PMID- 11057058 TI - Diabetic retinopathy screening among Hispanics in Lea County, New Mexico. AB - A project of diabetic retinopathy screening among Hispanics was implemented to increase awareness among the participants about diabetic retinopathy, screen for retinopathy, and make recommendations for further follow-up to enable them to save years of vision. Hispanics with diabetes are at a higher risk than non Hispanics to develop retinopathy. A convenience group of 19 adults participated in the project. A pretest to determine participants' knowledge about diabetic retinopathy was administered. Subsequently, the participants were screened for diabetic retinopathy. On that occasion, a nurse instructed the individual participants about different aspects of diabetic retinopathy and provided them with brochures. After three weeks, posttests were administered to evaluate their awareness level and compliance with recommendations. Data were analyzed using a dependent groups' t-test. A significant increase (6.6 percent) in the awareness levels of the participants was found. It is recommended that voluntary collaborative efforts among various individuals and institutions should occur to organize similar community projects that may otherwise become prohibitive due to costs. PMID- 11057059 TI - Using focus groups to design a community health program: what roles should volunteers play? AB - This volunteer-based community health advisory program illustrated the benefits of using focus groups to monitor and guide program development. Three focus groups, conducted by program evaluators, obtained input regarding the role of community volunteers. Focus group findings confirmed some of the assumptions held by program developers but refuted others. For example, prospective community participants and program developers were found to view the role of "volunteer" in the neighborhood health advisory program differently. As a result, adjustments were made in recruitment strategies and other procedural aspects to accommodate the residents' perspectives and values. This experience illustrated how preliminary focus group research can enhance linkage between program developers and community members. PMID- 11057060 TI - Improved indicators of infant mortality for integrated primary healthcare programmes. AB - Mortality and fertility rates are decreasing rapidly in many developing countries. It is argued that the indices commonly used as measures of these changes, i.e. infant mortality rate and fertility rate, ignore the interaction between mortality and fertility, and do not reflect their combined impact in lowering overall infant mortality. The paper proposes new indicators of infant mortality, termed fertility-adjusted infant mortality ratio (FIMR), age-specific, fertility-adjusted IMR (AFIMR), and total infant mortality ratio (TIMR) that are more sensitive to rapid demographic changes. These indicators include the combined effects of changes in both fertility and infant mortality rates on overall infant mortality in a region and appear to measure the effects of integrated health programmes better. Further, these conceptualize the mother infant pair as an appropriate unit with which to monitor mortality, and may be used for guiding allocation of resources intended to lower infant mortality. The application and usefulness of these indicators have been illustrated, using one hypothetical example and empirical data from the maternal-child health and family planning programme in Matlab, Bangladesh, as well as data from white and black population groups in the U.S.A. The results of these examples demonstrate that the new indicators are more sensitive than traditional measures when describing infant mortality, and may better reflect the perception in infant mortality status in the community. PMID- 11057061 TI - User-fees for family-planning methods: an analysis of payment behaviour among urban contraceptors in Bangladesh. AB - The study was carried out to review the experience with the existing user-fee (pricing) strategies and examine the socioeconomic and demographic factors associated with payment behaviour among contraceptors in urban Bangladesh for selected contraceptive methods, such as injectables, pill, and condom. Data for the study were drawn from a survey of more than 5,000 married women of reproductive age in Zone 3 of Dhaka city, Bangladesh, within the sample frame of the Urban Panel Survey of the ICDDR,B's former Urban MCH-FP Extension Project. The findings of the study showed that most (80%) urban contraceptors have been paying for selected family-planning services. This indicates the existence of a notable demand for contraceptives which suggests that there is scope for improved financial sustainability of the family-planning programme through charging appropriate user-fees for contraceptives with proper analyses of willingness-to pay among the contraceptors and price elasticities of demand. Higher socioeconomic status of households, marked by higher levels of education and house rent, and location of residence in non-slum areas, is predictive of paying for contraception. Households having 1-3 living child(ren) are also more likely to make payment for the selected contraceptive services. PMID- 11057062 TI - Poverty among widows of Kinshasa, Congo. AB - The linkages between poverty and death in the family in a sector of the city of Kinshasa, capital of the Democratic Republic of Congo (previously Zaire) were studied. The poor people have been identified using 3 convergent norms, described in the Methods of Materials section, based on total expenditure, calorie consumption in food, and proportion of expenditure for food. Family histories were recorded to understand the difficult situation of widow-headed households identified within the sample area. The relationship between death in the family and poverty was bi-directional: on the one hand, death of the breadwinner can accelerate the level of poverty; and on the other, poverty conditions can result in further deaths in the family. PMID- 11057063 TI - Relationship between feeding practices and weanling diarrhoea in northeast Thailand. AB - Diarrhoea is a major public health problem in Thailand. During November 1998 January 1999, a cross-sectional survey and a nested qualitative study were conducted to understand the relationship between feeding practices and weanling diarrhoea, and to describe the related local beliefs and practices in a subdistrict of northeast Thailand. A cluster-sampling method was used for selecting 156 weanlings aged 3-24 months. A structured interview was conducted with the main caregivers of these weanlings. The questionnaire used for the interview included items about feeding practices and diarrhoea-history of the weanlings in 2 months prior to the interview. Seven focus-group discussions with an opportunistic sample of the caregivers were held in the villages. A series of vignettes and unstructured questions were used for eliciting the local beliefs about weanling diarrhoea and its causes. Thirty-six (23%) of the 156 weanlings had diarrhoea in 2 months prior to the interview. The factors that were significantly related to reported weanling diarrhoea included consumption of unboiled water by weanlings (OR = 10, p = 0.03), not covering perishable foods (OR = 3, p = 0.02), and washing feeding utensils of weanlings without dishwashing detergent (OR = 3.1, p = 0.02), 'Su' and 'tongsia'--two common local terms--were used for describing different types of weanling diarrhoea. Many caregivers considered 'su' a natural occurrence in a child's development. The results suggest that some poor feeding practices may contribute to the higher risk of weanling diarrhoea in northeast Thailand. Some local beliefs about weanling diarrhoea may mask the true causes, and mislead messages about its prevention. PMID- 11057064 TI - Relationship of breast-feeding and hand-washing with dehydration in infants with diarrhoea due to Escherichia coli. AB - This prospective cohort study was carried out in the neonatal unit of the Yangon Children Hospital, Myanmar, to gather more information on the types of feedings and hand-washing practices of mothers as the determinant of severe dehydration in infants with acute diarrhoea due to Escherichia coli. The study subjects included 100 infants with diarrhoea, aged less than 4 months, admitted to the hospital from June 1997 to May 1998. Data on isolation of E. coli from rectal swab samples, types of feedings, hand-washing practices, and dehydration status were collected. Of the 100 cases, E. coli was isolated from rectal swab samples of 48 infants. Of these 48 cases, 28 had some dehydration and 20 had severe dehydration. Exclusive breast-feeding was observed only in the age group < 1 and > 1-2 month(s). The association of the severity of dehydration with other types of feedings compared to exclusive breast-feeding was not statistically significant. In this study, most mothers washed their hands with water only after cleansing their children's defaecation, and before and after feeding their children. The severity of dehydration was statistically significant in hand washing practices when compared to washing with water only and washing with soap and water. This study has shown the association between types of feedings and hand-washing practices with dehydration in infants with acute diarrhoea due to E. coli. The results of the study suggest that there is a need for appropriate intervention programmes to promote exclusive breast-feeding and hand-washing practices with soap and water after cleansing children's defaecation, and before and after feeding children. PMID- 11057065 TI - Perceived causes and management of diarrhoea in young children by market women in Enugu State, Nigeria. AB - The aim of the study was to determine the perceptions of mothers regarding the causes and management of diarrhoea of their children aged 0-24 months. In Enugu State, Nigeria, 80 market women whose children had diarrhoea during last 6 months were interviewed fortnightly. When possible, the children were observed to determine the types of diarrhoea and treatments given. Fifty-three of the women brought their children to market, and 27 left their children at home. Seventy-one percent of the mothers perceived that diarrhoea was caused by teething. The most common types of diarrhoea occurring in these children were watery diarrhoea (59%) and the so-called teething diarrhoea (29%). Dysentery (6%) and jedi jedi or frothy and mucoid stools (4%) occurred less frequently. In 68% of the cases, drugs were used alone or in conjunction with salt-sugar solution (SSS) or other forms of treatment. These drugs were prescribed by medical personnel (40%), patent medicine dealers (23%), or mothers themselves (30%). About 26% and 39% of the mothers treated, respectively, watery and teething diarrhoeas with drugs only, while 23% used SSS alone. The drugs used were mainly antimicrobials (34%) and a combination of antimicrobial, antimalarial, antacid, analgesic, and some local herbal preparations (21%). The results of the study showed the evidence of unnecessary use of drugs and ignorance about their potential adverse effects. These underscore the need for appropriate primary care education among the market women in Nigeria. PMID- 11057066 TI - Determinants of use rate of oral rehydration therapy for management of childhood diarrhoea in rural Bangladesh. AB - In rural Bangladesh, mothers were interviewed to identify factors that determine the use of oral rehydration therapy (ORT) for management of diarrhoea in children aged less than 5 years. The point prevalence of diarrhoea among 1,600 children was 11.6%, with 46% having acute watery diarrhoea. The overall ORT-use rate was 29%; only 17% of the cases used it adequately. Common reasons for not using ORS included misperception about diarrhoea and age of patients. Other reasons included incorrect assessments, severity, and difficulties with the administration of oral rehydration solutions. Promotion of ORT can be effected by improving the level of understanding of mothers with regard to assessment of severity, early initiation of treatment regardless of age, sex, type of diarrhoea, breast-feeding, and nutrition status. PMID- 11057067 TI - Intestinal amoebiasis: delayed-type hypersensitivity response in mice. AB - Delayed-type hypersensitive (DTH) response was evaluated in C3H/HeJ mice intestinally infected with Entamoeba histolytica. Infected and non-infected control mice were challenged with amoebic antigen on day 5, 10, 15, 20, 25, 30, 40, 50, and 60 post-infection. Maximum footpad swelling was observed after 24 hours of the challenge. The E. histolytica-infected mice exhibited a DTH response on day 5, 15, 20, 25, 40 and 60 post-infection. However, on day 10, 30, and 50, such response was similar to that of the non-infected control mice. The mice developed an evident DTH response late in the course of infection (25 days post infection). The infected mice did not show any alteration to their DTH response against heterologous unrelated antigen (sheep red blood cells), suggesting that cellular anergy was antigen-specific. PMID- 11057068 TI - Association of malarial parasitaemia with dehydrating diarrhoea in Nigerian children. AB - Records of 402 children--216 (53.7%) males and 186 (46.3%) females--aged 1-36 month(s), admitted to the Diarrhoea Treatment and Training Unit of the University of Benin Teaching Hospital, Benin city, Nigeria, during July 1993-June 1996, were reviewed to document the relationship between dehydration and malaria parasitaemia. There was a significant association between severity of dehydration and malaria parasitaemia (p < 0.0001). Association of parasitaemia (p < 0.006) with dehydration (p < 0.0001) was significantly more marked in patients with acute watery diarrhoea than in those with persistent and bloody diarrhoea. Parasitaemia was demonstrated in 50.5% of those not initially suspected to have malaria. Parasitaemia was also significantly associated with fever (p < 0.001) and fever coexisting with vomiting (p < 0.01). The prevalence of malaria associated diarrhoea was 61.7%. More infants (75.6%) than older children had diarrhoea. It was concluded that the prevalence of malaria-associated diarrhoea was high and that children with dehydration are more likely to manifest malaria parasitaemia. PMID- 11057070 TI - Clinical research and the office-based physician. PMID- 11057069 TI - Vitamin A deficiency in children with acute diarrhoea: a community-based study in Bangladesh. AB - The prevalences of nightblindness and xerophthalmia were assessed in 400 children, aged 6-59 months, with acute diarrhoea in a rural community in Bangladesh. The prevalences of nightblindness, conjunctival xerosis, and Bitot's spot were 7.8%, 9.5%, and 2.7% respectively. Fifty-two percent of the children who complained of nightblindness had ocular signs of vitamin A deficiency compared to 9% of those without nightblindness (p < 0.000). The nightblindness was significantly higher among the male children, aged 24-59 months, who were dysenteric and undernourished, did not consume vitamin A-containing foods daily, and were not breastfed. The coverage of periodic administration of vitamin A capsule was inversely related to the prevalence of nightblindness. This finding was determined by logistic regression analysis of data indicating that a combination of male sex, history of dysentery, absence of periodic administration of vitamin A treatment, and daily intake of vitamin A-containing foods gave the best-fitted model with an overall prediction of 92.5% of being nightblind. The findings of the study suggest that mothers should be educated to observe their diarrhoeal children about development of nightblindness and to seek treatment for it. The locally-relevant nutrition education should also be offered to them. PMID- 11057071 TI - "Traditional osteopathy": an oxymoron? PMID- 11057072 TI - Effects of a structured curriculum in osteopathic manipulative treatment (OMT) on osteopathic structural examinations and use of OMT for hospitalized patients. AB - Osteopathic manipulative treatment (OMT) is a defining feature of osteopathic medicine; however, use of OMT by osteopathic physicians is declining. Recent studies reveal that many osteopathic physicians are abandoning use of OMT as early as medical school. Current national efforts are aimed at reversing this trend by standardizing osteopathic medical records and clinical training in OMT. The authors found that a structured clinical curriculum in OMT taught to house staff significantly increased the percentage of patients who received osteopathic structural examinations and the percentage of patients who received OMT as part of their hospital care. PMID- 11057073 TI - Preparing medical students for the changing healthcare environment in United States. AB - Future graduates of medical schools in the United States will practice in healthcare environments increasingly predominated by managed care. Thus allopathic and osteopathic undergraduate and postgraduate residency training programs should begin to revise their respective curricula and conduct training in managed healthcare environments to prepare graduates for practice in managed care settings. The demand for curricular revision in medicine comes not only from prospective employers and government agencies, but from students and graduates. Educators, clinicians, and government officials have recently defined core competencies that are requisite to the education and preparation of future physicians through the work of the Council on Graduate Medical Education (COGME). This article discusses these core competencies and suggests strategies by which to implement them in undergraduate and graduate medical education. PMID- 11057074 TI - Osteopathic research imperative-III. 1936. PMID- 11057075 TI - The cranial bowl. 1944. PMID- 11057076 TI - Osteopathic cranial lesions. 1948. PMID- 11057077 TI - In COPD patients, body weight excess can mask lean tissue depletion: a simple method of estimation. AB - Although a great number of studies have been carried out on the relationship between chronic obstructive pulmonary disease (COPD) and low body weight, the identification of the most suitable warning signs of this wasting condition is still under debate. It has been indicated in earlier studies that body weight alone is of limited diagnostic value concerning this clinical condition in as far as a great number of COPD patients are usually overweight. For this reason, the aim of the current research was to find parameters that take into consideration the fact that body composition should be taken into account instead of weight only, and to assess whether COPD can be considered a "protein wasting disease", defining sensitive and significant indices of lean tissue depletion in relationship to the severity of the clinical symptoms. One hundred and seventy five stable COPD outpatients with differing degrees of bronchial obstruction and arterial blood gas abnormalities were consecutively recruited: anthropometric measurements and body composition analysis were carried out; 60 healthy subjects with normal pulmonary function, matched for sex, age and anthropometric parameters, were considered as controls. The data obtained showed a lower prevalence (9%) of underweight COPD patients in comparison with normal weight (37%) and overweight (54%) patients. In COPD patients, the phase angle measured by bioelectrical impedance analysis, whose deterioration is a good indicator of protein mass depletion, was altered by 19%, thus allowing the identification of currently malnourished subjects included in the overweight COPD patients group. In addition, significant correlations (p = 0.000) were found between the same nutritional variable, respiratory function and gas-exchange parameters, thus confirming that the more severe the stage of the pulmonary disease, the higher the degree of protein breakdown, regardless of body weight. PMID- 11057078 TI - Adequacy of oxygenation in a group of Danish patients with COPD on long-term oxygen therapy. AB - The aim of this study was to assess the adequacy of oxygenation in patients with chronic obstructive pulmonary disease (COPD) on long-term oxygen therapy (LTOT) in Copenhagen county. Twenty-six patients were enrolled and assessed prospectively by 24-h pulse oximetry. Patients comprised seven males and 19 females with a mean age of 69 +/- 5 yrs, arterial oxygen tension of 6.5 +/- 0.6 kPa, forced vital capacity of 1.47 L, forced expiratory volume in one second of 0.5 +/- 0.2 L (22 +/- 9% of predicted) and St. George's Respiratory Questionnaire total score of 61 +/- 14. The mean 24-h arterial oxygen saturation measured by pulse oximetry (Sp,O2) was within normal limits (94 +/- 1%), but mild hypoxaemia with a mean Sp,O2 of 89 +/- 4% and a total of 12 +/- 15 desaturation episodes per patient were observed, mostly during daily activities. Multiple regression analysis revealed no significant predictors for patient oxygenation. In summary, 24-h pulse oximetry showed adequate oxygenation in this group of COPD patients on LTOT, though mild hypoxaemia was present during some daily activities. PMID- 11057079 TI - Lung transplantation: the new point of view of an Italian centre. AB - Lung transplantation has become an accepted therapy for patients with end-stage lung disease. The survival rate after this operation is not, however, satisfactory, being 40-50% at 5 yrs after lung transplantation; infections and pulmonary rejection (acute and chronic) are the cause of this brief survival. Recently, it has been shown that lung transplantation is an advantageous solution only for selected pathologies. The introduction of alternatives to lung transplantation (lung volume reduction surgery in emphysema, prostacyclin therapy in primary pulmonary hypertension and pulmonary thromboendarterectomy in chronic thromboembolic hypertension) has modified the number of patients admitted to the lung transplantation waiting list. In this study, admission to the lung transplantation waiting list in the first 50 and in the following 50 months of activity of the Pulmonary Division Medical Centre of Montescano were retrospectively compared in order to verify whether experience gained has influenced admission to the lung transplantation waiting list. The mortality rate of patients with idiopathic pulmonary fibrosis (44%), chronic thromboembolic pulmonary hypertension (50%) and primary pulmonary hypertension (52%) before lung transplantation was high; the mortality after lung transplantation was low in idiopathic pulmonary fibrosis (16%), but rather high in primary pulmonary hypertension (55%) and chronic thromboembolic pulmonary hypertension (50%). In contrast, the mortality rate of patients with Eisenmenger's syndrome and emphysema was fairly low while on the lung transplantation waiting list, but rather high after lung transplantation. The trend in admission to the lung transplantation waiting list changed during the two observation periods, with a reduction in the number of patients with Eisenmenger's syndrome and emphysema, but not of those with idiopathic pulmonary fibrosis, chronic thromboembolic pulmonary hypertension and primary pulmonary hypertension. The experience gained modified the authors' approach to lung transplantation, but the "world" of lung transplantation still needs a lot more experience. PMID- 11057080 TI - Tuberculosis of the tongue secondary to pulmonary tuberculosis. AB - Both secondary and primary tuberculosis of the tongue and oral cavity are rare. A case of tuberculosis of the tongue secondary to pulmonary tuberculosis in a 41-yr old male patient is described. The clinical manifestation, diagnosis and response to the antituberculosis treatment are considered. PMID- 11057081 TI - Subpleural lung involvement in Behcet's disease: first localization of a systemic entity. AB - Behcet's disease (BD) is a chronic multisystem vasculitis, affecting many organs and the vascular system, of unknown aetiology. Eyes, skin, joints, the oral cavity, the central nervous system, and, less frequently, heart, lung, kidney, the genital system and the gastrointestinal tract can be involved. Intrathoracic manifestations of BD consist mainly of thromboembolism of the superior vena cava and/or other mediastinal veins; aneurysms of the aorta and pulmonary arteries; pulmonary infarct and haemorrhage; pleural effusion; and, rarely, myocardial and/or hilar lymphoid involvement. In the present case, the patient presented with BD with an asymptomatic subpleural lung mass and bilateral pulmonary artery enlargement. The patient was treated with a combination of surgical and medical therapy with complete resolution of the lung involvement and without any parenchymal relapses after an 8-month follow-up. PMID- 11057082 TI - Evaluation of tuberculosis treatment results in Italy, report 1998. Tuberculosis section of the National AIPO Study Group on Infectious Disease and the SMIRA Group. AB - In Italy no national data on tuberculosis (TB) treatment results were available. In 1995 the AIPO (Italian Association of Hospital Pneumologists) TB Study Group, in collaboration with the Istituto Superiore di Sanita (technical branch of the Ministry of Health), started a prospective monitoring activity based on World Health Organization (WHO) and International Union against Tuberculosis and Lung Disease (IUATLD) recommendations. Data were collected from a representative network of TB units nationwide, managing a significant proportion of all TB cases notified in Italy each year. The aim of this study was to analyse the case findings and treatment results for the year 1998. The number of TB cases reported was 1162 (61.7% males; 28.2% immigrants), 888 (76.6%) being new cases. Of these cases, 1,001 (86.4%) were pulmonary, 132 (11.4%) extrapulmonary and 26 (2.2%) both pulmonary and extrapulmonary. The main risk factors for TB were a history of recent contact, diabetes and alcohol abuse among native Italians and human immunodeficiency virus-seropositive status and history of recent contact among immigrants. The majority of immigrants were from Africa and South America, and had been in Italy > 24 months before diagnosis of TB. Forty-seven per cent (552 of 1,162) of patients had a positive direct sputum smear examination for alcohol acidfast bacilli; 30% were resistant to any drug (combined monoresistance (the sum of primary and acquired drug resistance) to rifampicin 2.3%; combined multidrug resistance 11.4%). In 97% of cases, the duration of treatment was < 12 months. The overall success rate (cured plus treatment completed excluding transferred out from the denominator) was 83%. A significantly higher percentage of deaths was found in native Italians (being age-related; p < 0.001), whereas immigrants had a higher default rate. PMID- 11057083 TI - Operative staging of lung cancer. AB - For earlier stage non-small cell lung cancer, surgical resection remains the most effective therapy. Complete resection of the primary tumour and lymph nodes should be the final aim in order to obtain the best long-term prognosis. Resectability depends on the tumour stage, and precise pre- and peroperative staging are of the utmost importance. In some cases, lung-sparing or extended operations are indicated. Despite modern scanning techniques, invasive staging by mediastinoscopy or thoracoscopy often remains necessary for determining resectability or deciding on a specific treatment. During thoracotomy, precise evaluation of the tumour and node factor is imperative for determining the extent of resection and achieving a complete tumour clearance. A systematic nodal dissection during thoracotomy is advised. Lung resection after induction therapy remains a technical challenge, especially after combined chemoradiotherapy. Peroperative staging is often difficult as distinction between viable tumour and fibrotic reaction is not easily made. Although combined modality treatment has an overall increased morbidity and mortality rate, it improves survival in selected cases of locally advanced non-small cell lung cancer. PMID- 11057084 TI - Is the evidence in favour of neoadjuvant chemotherapy in stage IIIA (N2) non small cell lung cancer solid enough? AB - In stage IIIA (N2) non-small cell lung cancer, single modality therapy with either surgery or radiation is curative in very few cases. The rationale behind neoadjuvant chemotherapy lies in the eradication of micrometastatic disease, which is often present when ipsilateral mediastinal or subcarinal lymph nodes (N2) are involved. Several studies have addressed the feasibility and efficacy of preoperative chemotherapy followed by surgery. All of these induction chemotherapy trials have reported a high radiographic response rate, high resectability rate and improved survival in completely resected patients. The findings reported in three published randomized trials indicate that the survival rate of stage IIIA patients is better with induction chemotherapy plus surgical resection than with resection alone. Phase II trials using preoperative concurrent chemoradiotherapy have been tested with encouraging results. Combined modality treatment in locally advanced non-small cell lung cancer continues to evolve and is the subject of ongoing research. New chemotherapeutic agents should be integrated into the neoadjuvant setting to improve results. A better understanding of the biology of tumours could well help in the optimization of these new agents. No doubt, in the future, molecular classification of non-small cell lung cancer will provide a useful tool for making therapy-related decisions. PMID- 11057085 TI - Therapeutic interventions aimed at reducing airway smooth muscle growth. AB - The airways of asthmatic subjects undergo complex changes which affect all compartments of the airway wall. The airway smooth muscle (ASM) is particularly affected and is increased in mass. Both hyperplasia and hypertrophy probably contribute to the altered mass of muscle which is a potential cause of airway hyperresponsiveness. The impedance to smooth muscle shortening resulting from the constitutive properties of the airway wall and the elasticity of the parenchyma may be more easily overcome by the excess muscle of the asthmatic airway. The ASM may also undergo important changes in its functional characteristics as a result of allergic sensitization and challenge. Increases in maximal shortening velocity, phospholipase C activity and membrane potential are some of the changes in the characteristics of sensitized ASM. The ASM may also change its phenotype from a contractile to a secretory tissue in response to the stimulus of airway inflammation and in doing so may contribute to the inflammatory process itself. The current therapies for asthma have the potential to counter some of the adverse effects of airway modelling, in particular in so far as the ASM is concerned. These effects should be kept in mind when considering the rationale for effective maintenance therapy for asthma. PMID- 11057086 TI - Airway fibrosis in asthma: mechanisms, consequences, and potential for therapeutic intervention. AB - A distinctive feature of asthma is the presence of a fibrotic response characterized by excess extracellular matrix deposition and proliferation of myofibroblasts, in the airway wall. This review discusses recent advances in the understanding of this phenomenon at the pathological and molecular levels, and its likely importance in the pathophysiology and symptomatology of the disease. Particular interest has centered on the potent fibrogenic molecule, transforming growth factor-beta (TGF-beta), and the evidence to link this molecule to airway fibrosis is discussed. Finally, the ability of existing anti-asthma treatments to influence the fibrotic response and the need to develop novel therapeutic strategies are considered. PMID- 11057087 TI - Mucus pathophysiology in COPD: differences to asthma, and pharmacotherapy. AB - Patients with chronic obstructive pulmonary disease (COPD) exhibit characteristics of airway mucus hypersecretion, including sputum production, increased luminal mucus, goblet cell hyperplasia and submucosal gland hypertrophy. These features are not common to all patients and the impact of hypersecretion on morbidity and mortality is a matter for debate. However, current evidence indicates that airway hypersecretion has pathophysiological and clinical significance in COPD, particularly as patients age or are prone to respiratory tract infection. This suggests that it is important to develop drugs that inhibit mucus hypersecretion in these patients. A number of drugs are currently available that may be of therapeutic benefit in hypersecretory disorders of the airways, e.g. glucocorticosteroids and anticholinergics. Novel compounds are undergoing preclinical research, e.g. inhibitors of epidermal growth factor receptor tyrosine kinase and antisense oligomers. However, preliminary data indicate that the mucus in COPD differs to that in asthma in that: 1) it is less viscous and without marked plasma exudation, 2) the ratio of mucin (MUC) 5AC:MUC5B may be reduced, and 3) there is full release of mucin into the airway lumen rather than "tethering" of mucus as in asthma. Consequently, future research should determine whether there really is an intrinsic abnormality specific to mucus in chronic obstructive pulmonary disease. Based upon this information, appropriate suppressers of mucus hypersecretion in chronic obstructive pulmonary disease can be developed. PMID- 11057088 TI - Prevention of respiratory syncytial virus infection with palivizumab. AB - Respiratory syncytial virus (RSV) infects virtually all children by the age of 2 yrs. Premature infants with chronic lung disease (CLD) are at risk of greater morbidity due to RSV infection. However, these infants represent a small proportion of all infants admitted to hospital with RSV infection, and hospitalization rates for this group appear to have decreased over the past decade. Prophylaxis against RSV infection has recently become available in the form of palivizumab, a humanized monoclonal antibody preparation. The IMpact trial demonstrated a 39% relative risk reduction in hospital admissions for RSV in cases in which palivizumab was administered to premature infants with CLD. However, palivizumab is a very expensive drug and cost-effectiveness analyses do not support its use in the majority of premature infants with or without CLD. The only group in which palivizumab should even be considered for use is premature infants with chronic lung disease at home on oxygen during their first respiratory syncytial virus season. In this group of infants, a detailed postlicensing audit needs to be performed to determine efficacy. PMID- 11057089 TI - Predictors of weanability. AB - It is generally accepted that mechanical ventilation should be discontinued at the earliest possible time in the course of a patient's illness. An empirical weaning approach exclusively based on clinical judgement without the recognition of predictors may cause prolonged mechanical ventilation. Therefore a variety of physiologic parameters have been investigated as so called "predictors" of weaning outcome. Weaning strategies exclusively based on predictors may miss the pathophysiological complexity underlying difficult weaning. Therefore, combining measurements of physiologically based parameters--performed during prescribed spontaneous breathing trials--and clinical judgement improve weaning outcome data. PMID- 11057090 TI - Noninvasive mechanical ventilation in hypercapnic respiratory failure: evidence based medicine. PMID- 11057091 TI - Flow/volume measurements from childhood to adulthood. AB - Flow/volume curves are the most frequently used pulmonary function test during childhood. Even preschool children are sometimes able to perform the maximal effort breathing techniques required for this pulmonary function test. The most important conditions for reliable measurement of flow/volume curves at all ages are reported here. In particular, testing atmosphere and equipment, pulmonary function technician requirements, testing procedure, reliability criteria, report making and interpretation are discussed. PMID- 11057092 TI - Dealing with the nanobacteria. PMID- 11057093 TI - Methods for detecting residues of cleaning agents during cleaning validation. AB - Cleaning validation procedures are carried out in order to assure that residues of cleaning agents are within acceptable limits after the cleaning process. Cleaning agents often consist of a mixture of various surfactants which are in a highly diluted state after the water rinsing procedure has been completed. This makes it difficult to find appropriate analytical methods that are sensitive enough to detect the cleaning agents. In addition, it is advantageous for the analytical methods to be simple to perform and to give results quickly. In this study, four different analytical methods are compared: visual detection of foam, pH, conductivity measurements, and analysis of total organic carbon (TOC). TOC was used as a reference method when evaluating the other three potential methods. The analyses were performed on different dilutions of the cleaning agents Vips Neutral, RBS-25, Debisan and Perform. The results demonstrated that the most sensitive method for analysis of Vips Neutral, Debisan and Perform is visual detection of foam, by which it is possible to detect concentrations of cleaning agents down to 10 micrograms/mL. RBS-25 was not detected below 200 micrograms/mL, probably because it is formulated with low-foaming surfactants. TOC analysis is less sensitive but has the advantage of being a quantitative analysis, while visual detection of foam is a semi-quantitative method. Visual detection of foam is easy to perform, gives a quick result, and requires no expensive instrumentation. The sensitivity of each method was found to be dependent upon the type of cleaning agent that was analyzed. PMID- 11057094 TI - Evaluation of a container closure integrity test model using visual inspection with confirmation by near infra-red spectroscopic analysis. AB - The objective of this study was to evaluate a novel test model involving an easy and rapid method to assess parenteral container/closure integrity. In this study, an extremely hygroscopic powder (methacholine chloride) was filled into the test vial/closure combination and served as an indicator of water vapor ingress into the package through either the stopper/glass interface and/or permeation through the closure. A visual means of detection was used initially, as the powder liquefies upon contact with a high-humidity environment. A further level of sensitivity was gained by using Near Infra-Red (NIR) spectroscopy to confirm that no additional water vapor was detectable in the test vials after being subjected to autoclave (worst-case water ingress) treatments. After two sequential autoclave cycles, none of the samples in the pilot study showed liquification of the indicator powder. This indicated that there was negligible ingress of water vapor, and therefore, the container/closure combination provided an adequate barrier to moisture ingress at the stress temperature and pressure conditions studied. The sensitivity of the NIR water ingress detection method was shown to be in the range needed for an acceptable vial integrity test. In conclusion, the model evaluated in this study can be used as an easy, rapid, and non-destructive closure integrity evaluation test. The use of such a NIR spectroscopy method would be immediately and directly amenable to the evaluation of vial integrity of dry powder-filled and lyophilized products, or can be used indirectly as shown in this study to assess container closure integrity for liquid-filled parenteral vial closure systems. PMID- 11057095 TI - The fate of silicone oil during heat-curing glass siliconization--changes in molecular parameters analyzed by size exclusion and high temperature gas chromatography. AB - The siliconization of pharmaceutical glass containers, usually for parenteral formulations, is performed in a so-called heat-curing process using diluted aqueous emulsions of medical grade silicone oils. To do this, the emulsion film is spread on the inner container surface, followed by an application of dry heat at temperatures above 300 degrees C. Water and surfactants are removed by degradation and vaporization, while the thermostable poly(dimethylsiloxane) (PDMS) is left on the surface. In the present study, heat-cured siliconized glass containers of two different types were solvent-extracted to obtain material of heat-treated PDMS. These samples were analyzed by size exclusion chromatography (SEC) and high-temperature gas chromatography (GC) with special respect to low molecular-weight siloxanes (LMWS). By comparison with the untreated starting materials, significant changes in the molecular weight distribution (MWD) of the silicone oil were revealed. Almost all of the LMWS present in untreated materials were not detectable in the heat-cured extract of a 100 cSt. Baysilone silicone oil. Small amounts of PDMS-molecules, with chain lengths of 25 up 45 siloxane units, were traceable. The examination of a second product of higher viscosity yielded unexpected results. The heat-treated extract contained none of the siloxanes that were detected in the starting material. Siloxanes of chain lengths of up to 45 units having molecular weights of over 3000 g/mol could not be found after the siliconization process. This led to the conclusion that not only vaporization effects must be responsible for their absence, but also that silicone suffers from a heat-induced degradation. The results of SEC and GC analysis were supported by each other. The whole molecular weight distribution and four distinct fractions were characterized by SEC, while the GC analysis was capable of a high-resolution view into the LMWS fraction below 3500 g/mol. In conclusion, the benefit of the heat treatment is that no LMWS, a source of toxicological concern, remain in the respective containers. On the other hand, an increase in molecular weight and viscosity of the silicone oil, and thus a possible change of the lubricating properties, is likely to happen through removal of LMWS. But these changes probably have no impact on the hydrophobic surface behavior of silicone-treated glass. PMID- 11057096 TI - The effect of media composition on the thermal resistance of Bacillus stearothermophilus. AB - Spores of Bacillus stearothermophilus ATCC 7953 were developed at 62 degrees C on 32 media composed of various amounts of 11 components: D-glucose, L-glutamic acid, yeast extract, peptone, sodium chloride, magnesium sulfate, ammonium phosphate, potassium phosphate, calcium chloride, ferrous sulfate and manganese sulfate. Statistical models were used and demonstrated a strong interaction of yeast/peptone/ammonium phosphate, contributing positively to the best sporulation yield (6-7 log10 spores). The most influential medium components on the thermal resistance (at 121 degrees C) of spores in suspension (calcium acetate, pH 9.7) were yeast extract (positively) and potassium phosphate (negatively), both creating the positive interaction, for spores from a 6-day incubation period. However, the strong negative effect of sodium chloride decreased the D-value from 1.81 min to 0.57 min upon increasing the incubation period (62 degrees C) from 3 days to 6 days. The D-glucose and peptone exhibited greater effects than the yeast extract and potassium phosphate interaction on D-values for 3-day spores on strip, just as the highly joint-positive peptone/sodium chloride effect maintained the thermal resistance of 6-day spores on strips. The spores on strip system showed less stability than the spores in suspension. The most stable spore system confirmed D-values at 121 degrees C at a range between 1.5 min and 1.9 min, which were obtained by keeping sodium chloride and potassium phosphate at minimum concentrations and yeast extract and peptone at maximum concentrations, regardless of the 3- to 6-day sporulation. PMID- 11057097 TI - Moist-heat sterilization in the UK pharmaceutical industry. The Authorised Persons (Sterilizers) Group UK. PMID- 11057098 TI - Reading the 'Net, revitalizing your mind. PMID- 11057099 TI - [Validation of the Polish version of St. George's respiratory questionnaire in patients with bronchial asthma]. AB - St George's Respiratory Questionnaire (SGRQ) has been widely used in the assessment of health related quality of life (HRQOL) in patients with asthma and chronic obstructive pulmonary disease. Introduction of the new language version of the HRQOL questionnaire needs to be preceded by a highly structured process of validation. We aimed to validate the Polish version of SGRQ in the group of 83 patients with asthma. Following the comprehension study, we thus evaluated reliability, validity, reproducibility, responsiveness, and measurement equivalence of the Polish version of SGRQ. Disease severity and health status were also concurrently assessed. The reliability was good, with Cronbach's alpha coefficient exceeding 0.75 for global and all subscale scores. There have been highly significant correlations between spirometric parameters, intensity of symptoms, health status self-assessment, and the degree of depression, and quality of life scores. Reproducibility, stability and responsiveness were confirmed in the follow-up study. Minimal clinically important difference was found to be 5.3 points. Polish version of SGRQ was found to be psychometrically equivalent to four other versions of SGRQ, which underscores its validity in the population of Polish asthmatics. PMID- 11057100 TI - [Secretion of interleukin-2 (IL-2) and interferon (IFN gamma)in whole blood cell culture stimulated with mitogens in patients with lung neoplasms]. AB - The aim of this study was to compare the ability of blood lymphocytes from lung cancer patients to secrete interleukin-2 (IL-2) and interferon gamma (IFNg) upon stimulation with mitogens with that of healthy donors. 42 patients with small cell lung cancer (SCLC), 30 patients with non small cell lung cancer (NSCLC) and 30 healthy donors were studied. The test was done in lung cancer patients before treatment. IL-2 and IFNg levels were measured with Elisa ready kits (Genzyme) in the supernatants of whole blood culture after stimulation with Pokeweed (PWM) and Phytohemagglutinin (PHA) mitogens. The results of the cytokine levels after stimulation were not normally distributed and thus were transformed to logarthms for statistical evaluation. The t-test for transformed results were used to asses the difference between groups. The median level of IFNg in the supernatant of whole blood cultures was significantly lower in lung cancer patients than in healthy blood donors both after PWM as well as after PHA stimulation. When patients with NSCLC and SCLC were regarded separately the lower level of IFNg in comparison with healthy donors was found in the supernatant of the blood cultures only after stimulation with PWM. The median level of IL-2 in the supernatant of whole blood culture in lung cancer patients was lower than in healthy blood donors only after PWM stimulation. The same was true for SCLC patients. In NSCLC IL-2 levels were significantly lower after stimulation with PWM as well after PHA stimulation. IN CONCLUSION: secretion of IL-2 and IFNg in whole blood culture after mitogen stimulation in lung cancer patients is significantly lower than in healthy donors. No significant differences between SCLC and NSCLC were found. PMID- 11057101 TI - [Analysis of motivation for cigarette smoking by teenagers based on a questionnaire distributed among school pupils staying in summer camps]. AB - Cigarette smoking by teenagers is conditioned by many factors with psychosocial ones among them. The aim of our study was to define the reasons for cigarette smoking by juveniles in Poland nowadays. The analysis was performed on the basis of questionnaire given to 598 school pupils resting in the summer camps in the Tricity of three neighbouring towns of Gdansk, Sopot and Gdynia. The study group included 357 females and 241 males, with an age of 8-19 years, on average 14.4. Out of the hole group 111 (18.6%) were smokers. This subgroup, 58 boys and 53 girls (age: 12-19 yrs.; on average 15.4), answered to several questions concerning cigarette smoking. Among remaining 487 non-smokers the prevalence of cigarette smoking among their parents was only assessed. As the main reason for smoking the majority (69.4%) of smokers reported the stress reduction. Pleasure was another frequently reported (29.7%) reason for cigarette smoking. Peers smoking was an important reason for cigarette smoking for 28.8% of school pupils. We found that the example of smoking parents significantly influenced the prevalence of smoking cigarettes among teenagers. The majority (73.9%) of smoking pupils knew the cigarette were harmful. Obtained results show the multiplicity of reasons for cigarette smoking by juveniles resulting mainly from psychosocial factors. There is a great necessity of antismoking programs for school pupils adjusted to specificity of psychology of adolescence as well as the exemplary role of adults creating a life style without cigarette. PMID- 11057102 TI - [Noninvasive intermittent positive pressure ventilation in treatment of chronic respiratory disease exacerbation]. AB - Noninvasive intermittent positive pressure ventilation (NIPPV) via nasal mask became a routine method of treatment of severe exacerbations of chronic respiratory failure. The aim of the study was to apply NIPPV in patients with COPD admitted to hospital due to exacerbation of the disease who on standard treatment developed progressing respiratory acidosis (pH < 7.30). Fourteen COPD patients were treated with NIPPV. Arterial blood gases at the beginning of treatment were: PaO2 41 +/- 9 mmHg, PaCO2 = 87 +/- 17 mmHg, pH = 7.30 +/- 0.05. In 10 patients NIPPV applied quasi continuously resulted in clinical improvement and an amelioration of arterial blood gases. PaO2 rose from 41 +/- 9 mmHg to 56 +/- 12 mmHg, PaCO2 fell from 85 +/- 17 to 57 +/- 9 mmHg and pH rose from 7.30 +/- 0.05 to 7.41 +/- 0.04. In 4 patients NIPPV did not prevent further progression of respiratory acidosis. They were intubated and mechanically ventilated. Three patients survived and were discharged home. One patient died from septic shock. We conclude that NIPPV is an effective method to treat respiratory acidosis developing during exacerbation of severe COPD. PMID- 11057103 TI - [Chemokine RANTES in bronchoalveolar lavage fluid(BAL)from two different lung segments indicated by high resolution tomography (HRCT) in patients with sarcoidosis]. AB - In 28 non-smoking patients with sarcoidosis (14 males, 14 females aged 19-51) the concentrations of cytokine RANTES were estimated in BAL fluid from two different lung segments: with the most (s.A) and with the least (s.B.) extensive involvement estimated by high resolution computed tomography (HRCT). In examined subjects 12 patients showed homogeneous distribution of HRCT changes (HD) in lung parenchyma and 16 showed nonhomogeneous distribution of HRCT changes (ND) with domination of pathological changes in upper lobes. Eleven healthy volunteers served as controls. In BALF from s.A and sB the significantly higher concentrations of RANTES were observed in comparison with control group (14.4 and 10.9 pg/ml vs 3.6 and 3.4 pg/ml respectively). In group (ND) in BALF from s.A (from upper lobes--the most occupied by HRCT changes) the concentrations of RANTES were significantly higher than in BALF from s.B (from lower lobes with the least involvement estimated by HRCT). RANTES concentrations in BALF from s.A and s.B positively correlated with lymphocytes count, lymphocytes CD3, CD4 and HLA DR+ and correlated negatively with diffusing capacity in sarcoid patients. Our results suggest the significant role in pathogenesis of sarcoidosis and in alveolitis process enhancement. PMID- 11057104 TI - [Primary and acquired drug resistance of tuberculosis bacilli in Poland]. AB - Information about resistant pattern of Mycobacterium tuberculosis isolates against antituberculon drugs is a very important part of tuberculosis control and indicates the directions of TB policy in each country. Poland joined WHO/IUATLD global project on drug resistance surveillance, and carried out the first prospective survey, simultaneously on primary and acquired drug resistance of tuberculosis patients according WHO/IUATLD recommendations. The programme covered the whole country, basing on cooperation between the National Reference Laboratory (NRL) with regional TB laboratories. Questionnaires and cultures were obtained from patients who excreted TB bacilli during the period from 1 November 1996 to 1 November 1997 (12 months). Drug susceptibility testing to INH, SM, EMB and RMP were performed on Lowenstein-Jensen medium according to the proportion method or/and radiometric Bactec 460 TB system. 3970 TB patients bacteriologically confirmed by culture were included in a one-year study. The male to female ratio was 2.6:1. Patients were at the age of 6 to 83 years. Majority of patients (86% males and 77% of females) was older than 35 years. Primary resistance to any drug was found in 3.6% of new cases and 2.4% of those patients who excreted monoresistant strains. No monoresistance to EMB was found. 18 patients (0.6%) were infected by MDR strains. Total resistance in new cases was for INH--2.6%, for SM--1.8%, for RMP--0.7% and for EMB--0.1%. Acquired resistance to any drug was found in 17.0% of treated. Majority of patients--7.7% excreted monoresistant strains. 7.0% were infected by MDR strains. Total resistance to INH was 14.8%, to SM--9.2%, to RMP--7.8%, and to EBM--2.5%. No correlation was found between sex and primary resistance rates. Among new cases, 3.7% of males and 3.3% of females were infected with resistant strains. However, among treated patients, males (20%) excreted resistant strains twice as much as females (9.1%). Mean age of women and men infected with primary and acquired resistant strains was similar. PMID- 11057105 TI - [Concentration of selected cytokines associated with monocytes/macrophages in serum of patients with different clinical stages of pulmonary tuberculosis]. AB - Monocytes/macrophages are the main effector cells as well as the essential elements in granuloma formation in tuberculosis. Serum concentration of some cytokines connected with monocytes/macrophages like IL-12, MCP-1, TNF-alpha i sTNFRI was assessed in patients with advanced bacteriologically confirmed pulmonary tuberculosis (I group) and in those with minimal tb changes, usually without clinical signs of the disease, and sputum smear-negative (II group). We found that patients from group I had significantly higher concentration of IL-12 and sTNFRI and lower level of MCP-1 in comparison to group II. Significant difference between groups of patients with tuberculosis and the control group was found only while assessing serum concentration of TNFRI. PMID- 11057106 TI - [Lung manifestation of visceral larva migration syndrome due to Toxocara canis infection]. AB - 32 year-old patient was hospitalized because of disseminated lung lesions. 2 years earlier he manifested chorioditis. Exact disease history suggested suspicion of toxocare infection. The diagnosis was confirmed by serological tests with anti-Toxocara canis antibodies, bronchial lavage and chest CT scan. Administration of 450 mg of dietylokarbamasin (Hetrazan) resulted in complete resolution of pulmonary lesions. PMID- 11057107 TI - [Pseudochylothorax during the course of rheumatoid arthritis]. AB - Psudochylothorax is uncommon among pleural fluids. It can be observed during tuberculosis or rheumatoid arthritis in majority. A case of a 62 years old man with chronic pleural fluid is presented. Patient had rheumatoid arthritis diagnosed 40 years ago. For last 13 years symptomsless bilateral pleural fluid was observed. Antituberculous drugs were used without success. Plural fluid obtained after puncture had high level of cholesterol with it[symbol: see text]s crystals, without chylomikrons and triglycerides. Diagnosis of pseudochylothorax in the course of rheumatoid arthritis was established. After plural puncture fluid was removed and did not appear later. Differential diagnosis of pleural fluids is presented. PMID- 11057108 TI - [Clinical significance of measuring levels of interleukin 2(IL-2) and it's soluble interleukin 2 receptor in serum of patients with lung cancer]. PMID- 11057109 TI - [The role of neutrophils in pathogenesis of chronic obstructive pulmonary disease]. PMID- 11057110 TI - [Perspectives for treatment of pulmonary fibrosis]. AB - Conventional therapy of fibrogenic lung diseases, based mainly on the use of glucocorticoids or immunosuppressive, cytotoxic agents, does not usually protect before progression of disease and accumulation of the excess connective tissue in lungs. This article is a review of potential new therapies in the treatment of pulmonary fibrosis. Novel therapies emerged from the use of animal models of pulmonary fibrosis and recent advances in the cellular and molecular biology of inflammatory reaction involve the use of substances directed against the action of pathogenic agents: growth factors, cytokines or oxidants. Thus, future therapeutic strategies will be aimed at more selective and effective methods of controlling the key elements of pulmonary fibrosis. PMID- 11057111 TI - [Cardiological state of offsprings of the mothers suffering from systemic lupus erythematosus]. AB - Systemic lupus erythematosus is a disease in which inflammatory process provoked by different antibodies affects many organs and systems. The circulatory system is one of them. In patients with systemic lupus erythematosus cardiac disorders are generally known and well proved. It is known that this disease has heritage background. Thus, the offsprings of patients suffering from systemic lupus erythematosus belong to a risk group. Moreover, it is thought that maternal antibodies crossing transplancentally to the fetus cause damages of tissues including the heart. The aim of this study was to evaluate cardiological status of 38 children whose mothers suffered from systemic lupus erythematosus. In all sick mothers the diagnosis fulfilled criteria of the American Rheumatism Association. The mothers have been remaining under medical treatment while the children have been under control and simultaneously prophylaxis of lupus has been undertaken. The study was undertaken in 17 girls and 21 boys aged 3 to 18 years (average: 12 +/- 4.5 years). Physical development of presented children was satisfactory. During cardiological examination all subjects were in good general condition, without any clinical evidence of collagen disease and infection. Obtained results were compared with the ones found in control group of 38 children of healthy mothers, being at the same age. Study methods were: physical examination, arterial blood pressure measurement, standard and 24 hours according to Holter method ECG record, echocardiographic and Doppler examination, and physical performance test according to Bruce's protocol. In children of sick mothers examined laboratory parameters were within the normal limits excluding the presence of antinuclear antibodies (controlled by indirect immunofluorescence test), result of which was positive in 15 studied children (39%). In the group of the children of sick mothers the abnormalities of sinus node function were detected in 12 cases (32%), significantly more often than in the control. There were found abnormalities of atrio-ventricular and intraventricular conduction in 15 subjects (40%). The premature beats of ventricular origin were noticed in 3 cases (8%). These disturbances were significantly different from the control group. In addition, correlation between the presence of antinuclear antibodies and the cardiac abnormalities was taken into consideration. So, significant correlation between antinuclear antibodies and heart rhythm disorders was proved. During echocardiographic examination structural and functional abnormalities were found. They were: ventricular septal defect (muscular part) (1), pericarditis effusion (1), prolapse of the mitral valve anterior leaflet (2), mitral valve regurgitation of the second degree (2) and increased diameter of left atrium (8). One girl from the studied group, suffering from atrio-ventricular block of III* was operated on because of persistent ductus arteriosus still in the newborn's period. At the same time the permanent pace-maker was implanted. After 1 year of age this girl was operated on because of atrial septal defect (ASD II). In studied group of children echocardiographic global indices of left ventricular systolic function were normal. The subclinical impairment of diastolic left ventricle function was found in 8 children with increased left atrium-aorta index. Both the theoretical knowledge and the results of the studies suggest that the offsprings of mothers suffering from SLE need a careful cardiological observation. PMID- 11057112 TI - [Assessment of the left ventricular function by tissue Doppler echocardiography]. AB - Myocardial velocity gradient is a new indicator of regional left ventricular contraction determined by a two dimensional tissue Doppler imaging technique. The main goal of this study was to compare myocardial velocity gradient in patients with old myocardial infarction and dilated cardiomyopathy to normal subjects. We assessed myocardial velocity gradient in 44 persons: 32 patients (19 men, 13 women, mean aged 51.2 +/- 6.1 years) and 12 healthy subjects (7 men, 5 women, mean age 49.3 +/- 8.3 years) who were divided into 4 groups. Group A--14 patients with old anteroseptal myocardial infarction, group B--7 patients with old posterior infarction, group C--11 patients with dilated cardiomyopathy and group D--12 healthy subjects. In normal subjects myocardial velocity gradient in the anteroseptal segment was mean 2.44 +/- 0.34 s-1 and in the posterior segment was 3.08 +/- 0.38 s-1. Myocardial velocity gradient in the infarct regions was significantly lower than in noninfarct regions as well as that in the corresponding regions in normal subjects. Gradient in the anteroseptal and posterior segments was in group A: 0.61 +/- 0.33 s-1 12.39 +/- 0.65 s-1, p < 0.001 respectively and group B: 2.11 +/- 0.45 s-1 10.91 +/- 0.34 s-1, p < 0.001 respectively. In patients with dilated cardiomyopathy gradient was significantly lower (nteroseptal segment 0.55 +/- 0.37 s-1, posterior segment 0.85 +/- 0.31 s 1) than that in normal subjects (p < 0.001). CONCLUSION: Myocardial velocity gradient is a new indicator for the quantitative assessment of regional left ventricular contraction. PMID- 11057113 TI - [Effect of arterio-venous fistula blood flow dynamics on ECG abnormalities in chronic hemodialysis patients]. AB - The dialysis doses is mostly dependent on well functional permanent vascular access. From the other hand high vascular access blood flow (Qva) may induce cardiac problems in HD patients. The purpose of this study was to investigate the effect of vascular access dynamics on electrocardiographic abnormalities in hemodialysis patients. Therefore, forty non-diabetic, HD patients, with native vascular access (VA) were divided into two equal groups; with Qva > 1500 ml/min (group A), and also Qva < 1500 ml/min (group B). The average of VA survival period was 28 +/- 18 (mean +/- SE) (group A), and 29 +/- 15 (months) (group B). The Qva measurements monitoring by color Doppler sonography included also: maximal velocity (Vmax), time average of maximal velocity (TAMX), pulsate index (PI), and resistive index (RI). Kt/V index was calculated, as classical parameter of adequacy, and also shunt recirculation using 3 urea samples was measured. For estimation of cardiac function we used M-mode echocardiography, and 24-hours ECG (Holter) monitoring. The occurrence of ventricular (VE), and supraventricular extrasystoles (SVE), ST-T, and ST characteristic as well were monitored by 24 hours Holter. CONCLUSIONS: 1. In the group with high Qva (A) we observed significantly higher number of VE, and also of SVE recorded by Holter monitoring compared with the low Qva group (B). 2. The mean number of patients with ST-T changes was higher in group A (12 vs. 7), but number of patients with recorded by Holter ST depression, and ST elevation between investigated groups were similar. 3. The mean number of ventricular arrhythmias of Lown classified as 4A, and 4B of Lown grading was significantly higher in the group with high Qva (A). PMID- 11057114 TI - [Quality of life assessment 3 months after surgical treatment of acquired heart valve disease]. AB - The quality of life in 114 patients with acquired heart valve disease 3 months after surgical treatment was estimated. The significant improvement of quality of life in the matter of physical, psychical and social factors after mitral, aortic and double valve replacement was noted. It was accompanied by increase of exercise capacity measured in 6-minute walk test and changing of NYHA functional classes. No correlation was found between age of patients, selected echocardiographic parameters and quality of life improvement. PMID- 11057115 TI - [Long-term results of coronary balloon angioplasty in various age groups]. AB - Some data indicate that natural history of coronary artery disease in younger patients is characterised by high dynamics and therefore the long-term results of revascularisation procedures have generally poorer outcome. To verify this we compared the early and the long-term results of balloon angioplasty in 630 consecutive patients divided into four age groups: < 40 years (77 patients), 41 50 years (247 patients), 51-60 years (160 patients) and > 60 years (146 patients). Groups differed significantly in many clinical factors: higher proportion of women and unstable angina were encountered in older groups, while higher frequency of hypertension, hypertriglycerydaemia, current smoking, familial history of angina, prior myocardial infarction, were more often observed in younger patients. Groups did not differ in such angiographic factors as: global ejection fraction (EF), presence of multivessel disease, type of dilated lesions and vessels, multilesion PTCA, except higher frequency of EF < 50% in patients < 40 years of age. Immediate results of angioplasty did not differ significantly between the respective age groups: success rate was 87-94%, complications rate between 4.5% and 6.5%, complete revascularisation was achieved in 46-61% patients (NS). In the mean 5-year follow-up period repeated angiography was carried out with comparable frequency in about half of the studied patients (NS). Restenosis rate equalled 21-42% and significantly increased with the patients' age (p = 0.02 in chi 2, 0.009 in log-rank test), the related reinterventions rate likewise (p = 0.05 in chi 2, 0.009 in log-rank test). We did not observe any differences among the respective groups with regard to significant atherosclerosis progression, which was encountered in 15-19% of patients (NS). Survival rate did not differ significantly either, being in fact quite high (96-99%). Myocardial infarction in follow-up significantly more frequently (p = 0.01) occurred in patients < 40 years of age, in comparison with patients > 60 years of age, although it did not differ significantly in terms of overall test for independence (p = 0.3) and log-rank test (p = 0.07). The frequency of major coronary events significantly increased according to patients' age as opposed to the event-free survival (p = 0.02 in both tests). Uni and multivariate analysis confirmed that age over 50 years is an independent factor of restenosis, reintervention, and major coronary event in follow-up. Patients functional status at the end of observation period, according to CCS criteria, proved that in the older age groups the percentage of patients with none, or minor anginal complaints decreased, whereas the proportion of patients exhibiting the symptoms of severe angina (Class III and IV) significantly increased (p = 0.006). CONCLUSIONS: Balloon angioplasty offered similar short-term outcome in all age groups, as well as the survival rate during the 5-year follow-up period. Frequency of restenosis significantly increased in older patients especially the ones over 50 years of age; this in turn resulting in a higher reinterventions rate among them. On the other hand, patients below 40 years of age suffered more frequently from myocardial infarction during the follow-up period. Major coronary events were more frequent in patients over 50 years of age. Better functional status was observed in younger patients at the end of observation period. PMID- 11057116 TI - [Personality and restenosis after percutaneous transluminal coronary angioplasty]. AB - The paper is an attempt to find out which personality traits predispose to increased risk of restenosis in patients after first percutaneous transluminal coronary angioplasty (PTCA). The Eysenck Personality Questionnaire-Revised (EPQ R) was used in 87 consecutive men (31 persons with university education, 26 secondary, and 20 occupational education; mean age 50 years, range from 32 to 72) on the second day after PTCA. Raw data were used in statistical analysis of EPQ R. Restenosis was identified in coronary angiography within several weeks to 6 months after PTCA. Restenosis was detected in 25 patients. Groups with and without restenosis were compared with respect to EPQ-R parameters referring to neurotism (14.8 +/- 3.26 vs. 12.4 +/- 5.63; p < 0.01), extroversion (13.7 +/- 3.82 vs. 13.7 +/- 3.95; NS) and psychotism (5.7 +/- 3.20 vs. 7.2 +/- 4.8; p = 0.08). There was positive correlation between the level of neurotism and the frequency of restenosis (logistic regression coefficient = 0.225; OR = 1.252; p = 0.03), but not with age and the level of education. In contrast, psychotism did not correlate significantly with frequency of restenosis. In conclusion, neurotism appears to affect the frequency of restenosis, which means that emotional imbalance through reducing immunity to stress and skills of effective coping with it increases the risk of restenosis. Psychological intervention directed at developing the skills of coping with stress should be a part of the therapy in patients after PTCA. PMID- 11057117 TI - [Quality of life in diabetic patients with coronary artery disease treated with percutaneous coronary angioplasty]. AB - Increased restenosis rate, higher incidence of coronary events, and, in some studies also increased mortality are observed during long-term follow-up in patients (pts) with diabetes mellitus treated with percutaneous coronary angioplasty. This is why some authors suggest that indications for PTCA in the group of diabetic pts should be significantly limited. The aim of our study was the estimation of clinical condition and quality of life in diabetic patients who underwent PTCA procedure in order to establish indications for percutaneous revascularisation in this group of pts. The study group consisted of 54 diabetic patients who were successfully treated with percutaneous coronary angioplasty in the period of 1987-1996. All pts were assessed clinically and quality of life was estimated on the basis of specially designed questionnaire. During mean 5-year follow-up 1(1.9%) patient died, 2 (3.7%) pts had acute myocardial infarction, restenosis was diagnosed in 25 (46.3%) pts. Repeated revascularisation was necessary in 27 (50%) pts. Significant clinical improvement was observed in the pts from the study group as compared to their clinical condition before the procedure (CCS 0 or I--61% vs 0%, p < 0.0001, III--9% vs. 39%, p < 0.0003, IV- 1.9% vs. 22%, p < 0.001). The rate of pts who were employed did not change in consequence of successful PTCA procedure. The number of pts returning to work was equal to the number of patients receiving social benefits. Subjective improvement was declared by 87% of pts. Lack of angina pectoris episodes was reported by 52% of pts and in 35% of pts the number of angina pectoris episodes was significantly reduced. Better tolerance of physical activity was declared by 59% of pts, and 65% of the studied pts fully came back to their non-professional activities. Partial return to non-professional activities was possible for 24% of pts. Normal sexual activity was declared by 65% of pts. Only 9% of the studied pts experienced significant limitation in their sexual activity. PTCA met the expectations of 89% of pts while the number of negative opinions, i.e. the lack of subjective improvement, impaired quality of life, or disappointment with the results of the PTCA procedure did not exceed 13%. Significantly worse results of the selected parameters describing the quality of life were observed in the group of diabetic pts treated with insulin. CONCLUSIONS: PTCA improved quality of life in 60-90% of pts with diabetes mellitus, nevertheless, did not affect the employment status of successfully treated pts. Patients who needed insulin therapy had worse indicators of life quality, however thorough analysis suggest that PTCA can be advised as a method of treatment in the group of diabetic patients with coronary artery disease. PMID- 11057118 TI - [The importance of advanced glycosylation end products in the creation and progression of atherosclerosis in non-insulin-dependent diabetes mellitus]. AB - Diabetic macroangiopathy is a major cause of morbidity and mortality of patients with non-insulindependent diabetes mellitus. Bad metabolic control of diabetes increases the risk of developing atherosclerotic complications. Hyperglycemia aids to increase the process of non-enzymatic protein glycosylation. Advanced glycation end products (AGEs) created in Maillard reaction play multidirectional role in creation of atherosclerosis in NIDDM. AGEs form in LDL, HDL and VLDL particles and increase their oxidative modification. They aid deposition of cholesterol and its esters in macrophages as well as creation of foam cells. Reaction of LDL-AGE particles with proteoglycans of blood vessel intima results in its thickening. LDL-AGE and ox-LDL take part in formation of late atherosclerotic changes influencing the expression of genes for cytokines and growth factors. AGEs promote prothrombotic changes. They block formation of nitric oxide which results in impaired vessel relaxation. They also take part in progressive vessel narrowing up to complete vessel occlusion by atheroslerotic plaque. PMID- 11057119 TI - [Percutaneous balloon valvuloplasty in stenotic bioprosthetic valves]. AB - There are only a few therapeutic indications that require implantation of bioprosthetic heart valves. To implant bioprosthesis is rare, because malfunction of the bioprosthesis occurs as a result of tissue degeneration in long-term follow up. The dysfunction is bioprosthesis insufficiency with or without stenosis. The degenerated bioprosthesis requires reimplantation, but every subsequent operation carries a higher risk. Stenotic bioprosthesis can be treated effectively by balloon valvuloplasty. The aim of the paper is to establish indications, coindications for the treatment, to present the complications and long-term follow up. PMID- 11057120 TI - [HMG-CoA reductase inhibitors in prevention of cardiovascular diseases: new mechanisms, aspects and trials]. AB - Use of lipid-lowering drugs in both primary and secondary prevention of cardiovascular disease (CVD) decreases significantly risk of myocardial infarction, stroke, incidence of cardiovascular events, reduces the cardiovascular mortality and morbidity as well as total mortality. HMG-CoA reductase inhibitors (statins) are most potent cholesterol-lowering drugs. Statins act by inhibition of HMG-CoA reductase activity, a rate--limiting step in synthesis of cholesterol and important metabolites of mevalonate--isoprenoids. The mechanisms by which favourable antiatherogenic actions of statins occur are complex. Statins inhibit proliferation and migration of vascular smooth muscle cells, reduce free-radicals generation and LDL modification, lower Lp(a) concentration, inhibit macrophage-derived foam cells accumulation and inhibit activation of platelets, thromboxane and PAI-1 synthesis. Use of statins in the therapy of hypercholesterolemia is presently recommended by NCEP, especially in high-risk groups (diabetes, post-CABG and PTCA, kidney and heart transplantation). Nevertheless, patients with CAD and moderately elevated LDL-C levels also benefit from the treatment with statins. Because of high costs of the therapy, statins of most favourable pharmacoeconomic profile should be used. PMID- 11057121 TI - [Antiphospholipid syndrome in valvular heart diseases, ischemic heart disease and vascular thrombosis]. AB - The antiphospholipid syndrome (APS) leads to venous and arterial thrombosis, cardiac diseases, neurological, gastroenterological and dermatological complications. The role of antiphospholipid antibodies in genesis of thrombi by interaction with plasma clotting factors is well known. There is no evidence of their influence on valvular heart diseases or atherogenesis. This paper presents views and opinions about APS and related cardiovascular complications. PMID- 11057122 TI - [Red wine in medicine: panacea, fashion or ... risk factor?]. AB - Red wine has been a subject of much interest of professionals representing different fields of medicine. However, most of scientific studies have been searching for the reason of so called French paradox, which means that in France and other mediterranean countries the morbidity and mortality due to ischaemic heart disease is significantly lower than in other developed countries, in spite of relatively high consumption of fat and saturated fatty acids. The cardio protective mechanism of red wine, although incompletely understood, is connected on one hand with the presence of ethanol which increases high-density lipoprotein cholesterol (HDL) and inhibits platelet aggregation, and on the other hand with the presence of polyphenols that have desirable biological properties. These include flavonoids, phenolic acids and stilbenes which are reputed to have antioxidant, vasorelaxing and antiplatelet properties. There is a considerable body of evidence indicating that regular consumption of red wine at moderate doses (200-400 ml a day) exerts a protective effect against ischaemic heart disease, other cardiovascular diseases, and perhaps diabetes, osteoporosis or some cancers. But, since alcohol intake involves a potential danger (risk of dependence, alcoholism, many organic diseases, migraine, allergies) medical recommendations of red wine consumption should be formulated very carefully. PMID- 11057123 TI - [Aicardi syndrome]. PMID- 11057124 TI - [Angelman syndrome]. PMID- 11057125 TI - [Cohen syndrome]. PMID- 11057126 TI - [Freeman-Sheldon syndrome, whistling face syndrome]. PMID- 11057127 TI - [Marden-Walker syndrome]. PMID- 11057128 TI - [Meckel-Gruber syndrome]. PMID- 11057129 TI - [Neu-Laxova syndrome]. PMID- 11057130 TI - [Pallister-Hall syndrome]. PMID- 11057131 TI - [Pallister-Killian syndrome]. PMID- 11057132 TI - [Parry-Romberg syndrome]. PMID- 11057133 TI - [Prader-Willi syndrome]. PMID- 11057134 TI - [Schinzel-Giedion syndrome]. PMID- 11057135 TI - [Schwartz-Jampel syndrome(chondrodystrophic myotonia)]. PMID- 11057136 TI - [Sjogren-Larsson syndrome]. PMID- 11057137 TI - [Steinert myotonic dystrophy syndrome]. PMID- 11057138 TI - [X-linked hydrocephalus syndrome]. PMID- 11057139 TI - [Cerebro-oculo-facio-skeletal (COFS) syndrome]. PMID- 11057140 TI - [Gardner syndrome]. PMID- 11057141 TI - [Goltz syndrome]. PMID- 11057142 TI - [Gorlin syndrome, basal cell nervus syndrome]. PMID- 11057143 TI - [Klippel-Trenaunay-Weber syndrome]. PMID- 11057144 TI - [Osler hemorrhagic telangiectasia syndrome]. PMID- 11057145 TI - [Proteus syndrome]. PMID- 11057146 TI - [Ruvalcaba-Myhre-Smith syndrome]. PMID- 11057147 TI - [Neurocutaneous melanosis, melanose neurocutanee]. PMID- 11057148 TI - [Dyskeratosis congenita (Zinsser-Cole-Engman syndrome)]. PMID- 11057149 TI - [Cutis marmorata telangiectatica congenita]. PMID- 11057150 TI - [Multiple lentigines syndrome, LEOPARD syndrome, Moynahan syndrome]. PMID- 11057151 TI - [Multiple neuroma syndrome/multiple endocrine neoplasia type 2B]. PMID- 11057152 TI - [Epidermal nevus syndrome]. PMID- 11057153 TI - [Retinoic acid embryopathy]. PMID- 11057154 TI - [Fetal aminopterin syndrome]. PMID- 11057155 TI - [Fetal alcohol effects]. PMID- 11057156 TI - [Fetal trimethadione effects]. PMID- 11057157 TI - [Fetal valproate syndrome]. PMID- 11057158 TI - [Fetal hydantoin effects]. PMID- 11057159 TI - [Fetal methylmercury syndrome]. PMID- 11057160 TI - [Fetal warfarin syndrome]. PMID- 11057161 TI - [Fetal varicella effects]. PMID- 11057162 TI - [Fetal rubella effects]. PMID- 11057163 TI - [Fetal iodine deficiency disorder]. PMID- 11057164 TI - [Maternal PKU fetal effects]. PMID- 11057165 TI - [Contiguous gene syndromes]. PMID- 11057166 TI - [X-linked mental retardation syndromes]. PMID- 11057167 TI - [Albright hereditary osteodystrophy]. PMID- 11057168 TI - [Antley-Bixler syndrome]. PMID- 11057169 TI - [Baller-Gerold syndrome]. PMID- 11057170 TI - [Bardet-Biedl syndrome(BBS)]. PMID- 11057171 TI - [Beckwith-Wiedemann syndrome]. PMID- 11057172 TI - [Berardinelli lipodystrophy syndrome]. PMID- 11057173 TI - [Bloom syndrome]. PMID- 11057174 TI - [Borjeson-Forssman-Lehmann syndrome]. PMID- 11057175 TI - [CATCH 22]. PMID- 11057176 TI - [CHARGE association: a review]. PMID- 11057177 TI - [Chediak-Higashi syndrome]. PMID- 11057178 TI - [CHILD syndrome]. PMID- 11057179 TI - [Clouston syndrome, hydrotic ectodermal dysplasia]. PMID- 11057180 TI - [Coffin-Lowry syndrome]. PMID- 11057181 TI - [Coffin-Siris syndrome]. PMID- 11057182 TI - [Darier's disease]. PMID- 11057183 TI - [de Lange syndrome]. PMID- 11057184 TI - [Dubowitz syndrome]. PMID- 11057185 TI - [Dyggve-Melchior-Clausen syndrome]. PMID- 11057186 TI - [Dyke-Davidoff-Masson syndrome]. PMID- 11057187 TI - [Fanconi pancytopenia syndrome]. PMID- 11057188 TI - [FG syndrome]. PMID- 11057189 TI - [Greig cephalopolysyndactyly syndrome]. PMID- 11057190 TI - [Hajdu-Cheney syndrome]. PMID- 11057191 TI - [Hallermann-Streiff syndrome]. PMID- 11057192 TI - [Harlequin syndrome]. PMID- 11057193 TI - [Jarcho-Levin syndrome]. PMID- 11057194 TI - [Johanson-Blizzard syndrome]. PMID- 11057195 TI - [Kallmann syndrome]. PMID- 11057196 TI - [Langer-Giedion syndrome(tricho-rhino-phalangeal syndrome, type II; TRPS II)]. PMID- 11057197 TI - [Lenz-Majewski hyperostosis syndrome]. PMID- 11057198 TI - [Leprechaunism(Donohue syndrome)]. PMID- 11057199 TI - [Marshall syndrome]. PMID- 11057200 TI - [Marshall-Smith syndrome]. PMID- 11057201 TI - [Mobius syndrome]. PMID- 11057202 TI - [Nail-patella syndrome]. PMID- 11057203 TI - [Noonan syndrome]. PMID- 11057204 TI - [Oculo-auriculo-vertebral spectrum]. PMID- 11057205 TI - [Opitz syndrome]. PMID- 11057206 TI - [Oromandibular-limb hypogenesis spectrum]. PMID- 11057207 TI - [Papillon-Psaume syndrome]. PMID- 11057208 TI - [Rett syndrome]. PMID- 11057209 TI - [Rieger syndrome]. PMID- 11057210 TI - [Roberts-SC phocomelia syndrome]. PMID- 11057211 TI - [Robinow syndrome]. PMID- 11057212 TI - [Ross syndrome]. PMID- 11057213 TI - [Rubinstein-Taybi syndrome]. PMID- 11057214 TI - [Ruvalcaba syndrome]. PMID- 11057215 TI - [Seckel syndrome]. PMID- 11057216 TI - [Senter syndrome]. PMID- 11057217 TI - [Shprintzen syndrome]. PMID- 11057218 TI - [Smith-Lemli-Opitz syndrome(#270400)]. PMID- 11057219 TI - [Waardenburg syndrome]. PMID- 11057220 TI - [Weaver syndrome]. PMID- 11057221 TI - [Wildervanck syndrome, cervico-oculo-acoustic syndrome]. PMID- 11057222 TI - [Williams syndrome(Williams-Beuren syndrome), elfin face syndrome]. PMID- 11057223 TI - [Wyburn-Mason syndrome]. PMID- 11057224 TI - [Arima syndrome(cerebro-oculo-hepato-renal syndrome)]. PMID- 11057225 TI - [Kabuki make-up syndrome]. PMID- 11057226 TI - [Blepharophimosis, epicanthus inversus, and ptosis syndrome(BPES)]. PMID- 11057227 TI - [Oral-facial-digital syndrome]. PMID- 11057228 TI - [Cleido-cranial dysplasia]. PMID- 11057229 TI - [Rhizomelic chondrodysplasia punctata]. PMID- 11057230 TI - [Distichiasis-lymphedema syndrome]. PMID- 11057231 TI - [Acrodysostosis]. PMID- 11057232 TI - [Congenital insensitivity to pain with anhidrosis, Nishida syndrome]. PMID- 11057233 TI - [Short rib-polydactyly syndrome, Majewski type]. PMID- 11057234 TI - [Thanatophoric dysplasia]. PMID- 11057235 TI - [Arteriohepatic dysplasia]. PMID- 11057236 TI - [Chondroectodermal dysplasia, Ellis-van Creveld syndrome]. PMID- 11057237 TI - [Achondroplasia, hypochondroplasia]. PMID- 11057238 TI - [Cerebro-costo-mandibular syndrome]. PMID- 11057239 TI - [Camptomelic dysplasia]. PMID- 11057240 TI - [4p deletion syndrome]. PMID- 11057241 TI - [Trisomy 4p syndrome]. PMID- 11057242 TI - [Cat cry syndrome]. PMID- 11057243 TI - [Trisomy 8 mosaic syndrome]. PMID- 11057244 TI - [9p-syndrome]. PMID- 11057245 TI - [9p trisomy syndrome]. PMID- 11057246 TI - [Trisomy 9 mosaic syndrome]. PMID- 11057247 TI - [Partial trisomy 10q syndrome]. PMID- 11057248 TI - [Aniridia-Wilms tumor association]. PMID- 11057249 TI - [Monosomy 13q syndrome]. PMID- 11057250 TI - [Trisomy 13 syndrome, Patau syndrome]. PMID- 11057251 TI - [18p-syndrome]. PMID- 11057252 TI - [18q-syndrome]. PMID- 11057253 TI - [Trisomy 18 syndrome(Edwards syndrome)]. PMID- 11057254 TI - [20p trisomy syndrome]. PMID- 11057255 TI - [Down syndrome]. PMID- 11057256 TI - [Cat eye syndrome]. PMID- 11057257 TI - [Turner syndrome]. PMID- 11057258 TI - [Sex chromosome abnormality]. PMID- 11057259 TI - [Klinefelter syndrome]. PMID- 11057260 TI - [Fragile X syndrome]. PMID- 11057261 TI - [Triploidy syndrome and diploid/triploid mixoploidy syndrome]. PMID- 11057262 TI - [HELLP(hemolysis, elevated liver enzymes, low platelet count) syndrome]. PMID- 11057263 TI - [Langerhans cell histiocytosis]. PMID- 11057264 TI - [Melkersson-Rosenthal syndrome]. PMID- 11057265 TI - [Sneddon's syndrome]. PMID- 11057266 TI - [Apparent death]. PMID- 11057267 TI - [Mountain sickness]. PMID- 11057268 TI - [Antiphospholipid syndrome(APS)]. PMID- 11057269 TI - [Eclampsia]. PMID- 11057270 TI - [Fat embolism syndrome]. PMID- 11057271 TI - [Neurosarcoidosis]. PMID- 11057272 TI - [Neurological symptoms and signs in electrolytic imbalance]. PMID- 11057273 TI - [Superficial siderosis of the central nervous system]. PMID- 11057274 TI - [Membranous lipodystrophy]. PMID- 11057275 TI - [Charcot-Marie-Tooth disease]. PMID- 11057276 TI - [Hereditary sensory and autonomic neuropathy(HSAN)]. PMID- 11057277 TI - [Hereditary pressure sensitive neuropathy]. PMID- 11057278 TI - [Giant axonal neuropathy]. PMID- 11057279 TI - [Peripheral neuropathy associated with spinocerebellar ataxia]. PMID- 11057280 TI - [Neonatal hypoxic-ischemic encephalopathy]. PMID- 11057281 TI - [Intracranial hemorrhage]. PMID- 11057282 TI - [Hypoglycemia and brain injury]. PMID- 11057283 TI - [Bilirubin and brain injury]. PMID- 11057284 TI - [TORCH complex]. PMID- 11057285 TI - [HIV infection]. PMID- 11057286 TI - [Bacterial meningitis]. PMID- 11057287 TI - [Neonatal fungal infections]. PMID- 11057288 TI - [Mechanical birth injury]. PMID- 11057289 TI - [Fahr (idiopathic symmetrical cerebral calcification)]. PMID- 11057290 TI - [Hallervorden-Spatz disease]. PMID- 11057291 TI - [Infantile bilateral striatal necrosis]. PMID- 11057292 TI - [Intravascular malignant lymphomatosis (IML)]. PMID- 11057293 TI - [Kaplan-Grumbach-Hoyt syndrome]. PMID- 11057294 TI - [Non-progressive familial congenital cerebellar hypoplasia]. PMID- 11057295 TI - [Posterior cortical atrophy]. PMID- 11057296 TI - [Reye's and Reye's-like syndromes]. PMID- 11057297 TI - [Posttraumatic spinal pseudomeningocele]. PMID- 11057298 TI - [Exencephaly]. PMID- 11057299 TI - [Familial idiopathic brain calcification]. PMID- 11057300 TI - [Tension pneumocephalus]. PMID- 11057301 TI - [Bobble-head doll syndrome]. PMID- 11057302 TI - [Acute necrotizing encephalopathy of childhood]. PMID- 11057303 TI - [Neonatal progeroid syndrome]. PMID- 11057304 TI - [Pancreatic encephalopathy]. PMID- 11057305 TI - [Hydrocephalus]. PMID- 11057306 TI - [Normal pressure hydrocephalus(NPH)]. PMID- 11057307 TI - [Pseudoxanthoma elasticum(PXE)]. PMID- 11057308 TI - [Low cerebrospinal fluid pressure syndrome]. PMID- 11057309 TI - [Postanoxic-ischemic encephalopathy]. PMID- 11057310 TI - [Uremic neuropathy, uremic encephalopathy]. PMID- 11057311 TI - [Radiation-induced nervous system injury]. PMID- 11057312 TI - [Prevention and treatment of the side effects of cancer chemotherapy]. AB - Management of the side effects of chemotherapy in cancer patients is important because side effects can affect the tolerability and continuation of therapy, in addition to lowering the quality of life of patients. A significant efficacy of serotonin receptor antagonists against nausea and vomiting due to cancer chemotherapy, and granulocyte colony-stimulating factor against neutropenia secondary to chemotherapy, has been recently demonstrated. New chemoprotective drugs have been developed, such as amifostine for cisplatin-induced nephrotoxicity and neurotoxicity, and dexrazoxane for cardiac toxicity due to anthracyclines. Antiviral agents including lamivudine and interferons suppress virus replication, preventing the development of fulminant hepatitis during chemotherapy in cancer patients who have chronic hepatitis B infection. Various supportive therapies have resulted in the advance of cancer chemotherapy. PMID- 11057313 TI - [Oral cancer chemotherapy in the clinic]. AB - The present status of oral chemotherapeutic agents and the treatment of cancer were reviewed. Twenty anticancer agents are commercially available in Japan excluding hormonal and BRM agents. Cancers for the outpatient chemotherapy are limited because useful drugs are few for such circumstances. Now new potent agents such as S-1, capecitabine and molecular targeting are available. This kind of chemotherapy is needed for growing number of elderly patients for QOL and medical cost. PMID- 11057314 TI - [The development of an outpatient treatment system for cancer chemotherapy]. AB - It is necessary to develop a system of treatment to provide cancer chemotherapy on an outpatient basis. Facilities needed include treatment service sites, outpatient laboratories, and chemotherapy pharmacies where safety cabinets are available. Medical oncologists, chemotherapy nurses and pharmacists should conduct cancer chemotherapy after obtaining informed consent from the patient. Chemotherapy protocols, ordering systems and risk management for medical errors must also be established in the agreement to perform chemotherapy. The problems encountered in the experiences at the Outpatient Treatment Service of National Cancer Center Hospital East are discussed. PMID- 11057315 TI - [Outpatient clinic in Saitama Cancer Center]. AB - The achievements of the Outpatient Clinic Center in Saitama Cancer Center from April 1999 to March 2000 were reanalyzed, and their characteristics are discussed. Outpatients used the Outpatient Clinic Center a total of 9,862 times. The treatments and procedures were 923 blood transfusions, 4,308 infusions without anticancer drugs, and 4,409 consultations from outpatients. In addition, a total of 4,036 chemotherapies were performed in this outpatient setting. Fifty seven percent of the chemotherapies done by the Pulmonary Section contained the new agents of docetaxel, irinotecan, vinorelbine and gemcitabine (206, 187, 72 and 135 times, respectively). Irinotecan began to be used in outpatient settings after the use of the Oral Alkalization and Control of Defecation was initiated (Proc ASCO 18: 492a, 1999). These drugs were safely given to outpatients. No paclitaxel was used in our Outpatient Clinic Center because of the difficulty of its premedication in Japan, indicating the necessity of development of a new and simple premedication. PMID- 11057316 TI - [Outpatient treatment for gastrointestinal tract cancer in the Department of Surgery]. AB - Recently, outpatient cancer treatment has been rapidly increasing in order to cut medical expenditures and promote the QOL of cancer patients. However, the efficacy of outpatient cancer treatment is still uncertain. When performing outpatient cancer treatment, one must have a clear idea of the expected effect, and provide appropriate treatment so that none of the expected benefit is sacrificed for the improved convenience. PMID- 11057317 TI - [Radiotherapy for outpatients]. AB - Radiotherapy can be done on an outpatient basis depending on the patient's cancer stage, radiation method, concurrent therapy, treatment purpose and general condition. Daily treatment time takes only about 1 minute, but the patient has to come 5 times a week for about 6 weeks for radical treatment and about 3 weeks for palliative treatment. If treated with radiotherapy alone, most patients suffer no severe adverse effects in their daily life. Therefore, radiotherapy can be done on an outpatient basis only if the patient is able to come frequently to the clinic for several weeks. Breast cancer patients commonly undergo radiotherapy in the outpatient clinic, such as breast conserving therapy, postoperative therapy and radiotherapy for local recurrence after mastectomy. Prostate cancer is also commonly treated with radical radiotherapy, postoperative therapy and radiotherapy for local recurrence after prostatectomy. Usually, postoperative irradiation for other cancers and radiotherapy for bone metastases are also undergone on an outpatient basis. During outpatient treatment, it is important to predict and avoid severe normal tissue reactions to radiotherapy, such as myelosuppression and/or pneumonitis, before they are apparent by watching the patient carefully. PMID- 11057318 TI - [Hospice and palliative care in the outpatient department]. AB - In the medical environment, information disclosure to patients and respect of autonomy have spread rapidly. Today, many terminally-ill cancer patients wish to spend as much time at home as possible. In such situations the patient who has been informed that curative treatments are no longer expected to be beneficial can now hope to receive home care and visiting care from hospice/palliative care services. The essential concepts of hospice/palliative care are symptom management, communication, family care and a multidisciplinary approach. These concepts are also important in the outpatient department. In particular, medical staff need to understand and utilize management strategies for common symptoms from which terminally-ill cancer patients suffer (ex. cancer pain, anorexia/fatigue, dyspnea, nausea/vomiting, constipation, hypercalcemia and psychological symptoms). They also need to know how to use continuous subcutaneous infusion for symptom management in the patients last few days. The present paper explains the clinical practices of hospice/palliative care in the outpatient department. Also discussed is support of individual lives so that maximum QOL is provided for patients kept at home. PMID- 11057319 TI - [Development of molecular targeting drugs for the treatment of cancer-therapeutic potential and issues to be addressed in global development]. AB - A survey of cancer treatment in a sample of hospitals > 100 beds conducted in 1998 compared with experience in the US showed that good progress has been achieved in Japan in the screening and early treatment of gastric cancer, and that the prognosis for breast cancer is better than in the West. Although in the past, the cytotoxic therapies available to physicians in Japan vs the West have been different, recent acceleration of regulatory review will result in a convergence of treatment paradigms and some improvement in acute response in many tumour types. However, world wide there is a need for new improved therapies in all cancers evaluated. Particular needs are in the management of NSCLC, advanced disease and cancers which form micrometastases. The eventual hope is that cancer can be turned from a lethal disease into a chronic disease where patients maintain a good QOL. Apart from anti hormonal therapies, the usual approach has been to kill the cancerous cells. However, the new approaches to intervening in the growth and migration of cancerous cells or the host tissue response by molecular targeting offer the promise of achieving a step change in therapy. Although EGF tyrosine Kinase inhibitors such as ZD 1839 have been shown to cause a conventional tumour response in NSCLC, many of these new approaches are unlikely to show a short term response even if they have the capacity to affect tumour development and increase disease free survival. Some compounds will require combination therapy with a conventional cytotoxic or radiotherapy to show their full benefit. For conventional cytotoxics, the usual approach to development has been to select the maximum tolerated dose and then evaluate the efficacy in advanced disease. However, for the new approaches which will not have such severe dose limiting toxicities, it will be necessary to select a surrogate marker of the intended biological effect to select the optimal biological dose (OBD) and dose regimen in phase I/II studies for further evaluation in phase II or III studies which are designed to show the expected patient benefit. The tumour target, the stage of the disease and the possible need for concomitant therapy will also have to be considered according to the mechanism of action of the product. PMID- 11057320 TI - [Treatment and prognosis of children with relapsed non-Hodgkin's lymphoma--a report from CCLSG-NHL 890 Study. Children's Cancer and Leukemia Study Group (CCLSG)]. AB - To address the issue of salvageability in relapsed children with NHL who had all received the same frontline therapy, we retrospectively studied the treatment response and the outcome of 27 children who relapsed following the CCLSG-NHL890 protocol. The reinduction rates and 3-year survival rates (mean +/- SD) were as follows: lymphoblastic lymphoma (LB, n = 9), 44% & 17 +/- 14%; leukemia lymphoma syndrome (LLS, n = 8), 25% & 0%; large cell lymphoma (LC, n = 3) 100% & 67 +/- 27%; Burkitt's lymphoma (B, n = 7) 0% & 0%. Thus, the salvageability of LC lymphoma was good, but the outcome of Burkitt's lymphoma was very poor. CCLSG NHL960 protocol for LB lymphomas and intensive multiagent regimens for LC lymphomas produced favorable response rates, but the effect of the high-dose Ara C regimen for Burkitt's lymphoma was not determined. The initial stages of the disease seemed to be associated with the patient outcome: the outcome of the patients in stage IV was inferior to that of patients in stages II or III. Other clinical variables, such as relapse sites, relapse time and BM rescue did not affect the patients' outcome. PMID- 11057321 TI - [Weekly administration of paclitaxel for advanced or metastatic breast cancer- short-course premedications for outpatients]. AB - A phase II trial has demonstrated that paclitaxel (210 mg/m2/3 hr) showed a 33.3% response rate among anthracycline-resistant breast cancer patients in Japan. Recently, weekly dosing of paclitaxel has been demonstrated to be a well tolerated, feasible and effective administration schedule. Standard premedication is commonly administered prior to treatment with paclitaxel. However, this regimen requires dexamethasone administration beginning at 12 to 14 hours prior to paclitaxel, which would not be convenient for outpatients. In this study, paclitaxel was administered by 1 hour intravenous infusion at a dose of 80 mg/m2 every week. Administration was continued for 3 weeks with a 1 week rest. A short course premedication schedule consisted of dexamethasone 20 mg i.v. (diluted in 50 ml normal saline, 1 hour prior to paclitaxel), and oral diphenhydramine 50 mg, H2-antagonist and anti-emetic agent i.v. (diluted in 50 ml normal saline, 30 minutes prior to paclitaxel). A total of 14 outpatients were enrolled in the study. There were 10 partial responders and no complete responders, and the overall response rate was 71.4%. No hypersensitivity reactions were observed, and grade 3/4 leukopenia occurred in 43% (6/14). Allopecia was observed in 4 patients, and peripheral neuropathy in 1 patient (both grade 1). Weekly administration of paclitaxel is effective and well-tolerated in patients with advanced or metastatic breast cancer, with a minimum of peripheral neuropathy. In addition to the above, no hypersensitive reaction in the short course premedication schedule suggests that this administration schedule is feasible for outpatients. PMID- 11057322 TI - [A combined effect of fadrozole and tamoxifen in postmenopausal patients with recurrent breast cancer: a preliminary report: Japanese Cooperative Study Group of Fadrozole and Tamoxifen]. AB - To elucidate the combined effects of fadrozole (nonsteroidal aromatase inhibitor) and tamoxifen, 11 postmenopausal patients with recurrent breast cancer were examined between October 1996 and June 1998. One patient, 49 years old, was ineligible due to the short period after castration. The patients were aged 53-71 years (mean 63.5). PS was 0-1. Six patients were pre-treated with tamoxifen and 6 with oral 5-FU derivatives. One had no previous treatment. The target lesions were soft tissues in 5, bone in 4, lungs in 6 and liver in 1. The response was CR in 2, PR in 2, SD (longer than 24 weeks) in 2, NC in 1 and PD in 3. Consequently, the response rate was 60% (6 out of 10 eligible cases). Hormonal concentration was measured before and after administration of two drugs in weeks 2, 4, 6, 8 and at the end of the treatment, and significant decreases in estrogens in peripheral blood were observed. Adverse effects (4 cases of low grade headache, dizziness and elevation of GOT, GPT, gamma-GTP) did not influence the continuous administration of the drugs. We conclude that combined administration of fadrozole (2nd generation aromatase inhibitor) and tamoxifen produces a good response in postmenopausal recurrent breast cancer patients, and can be a useful treatment for patients with breast cancer. PMID- 11057323 TI - [A preliminary study on dihydropyrimidine dehydrogenase activity in breast cancer]. AB - We studied dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS), key enzymes in regulating the pharmacokinetics and chemosensitivity to 5-FU, in 36 breast cancer patients as a control group and 18 patients as a 5-FU group, in which 5-FU was given orally for 2 weeks before surgery at a dose of 200 mg/day. Cancer tissues with adjacent normal tissue were sampled and stored until the assay. The DPD activity and TS amount were determined according to the radio enzymatic assay and radiobinding assay, respectively. The DPD activity was significantly higher in breast cancer than in the adjacent breast tissue. This finding was observed in T1 and T2, node negative and ER positive breast cancers in the control group as well as in the 5-FU group. The DPD activity in T3 was significantly lower than in T1, and that of the adjacent breast tissue in T3 was significantly higher than in T1; therefore, the tumoral/non-tumoral ratio of DPD activity was significantly lower in T3 than T1. TS was significantly elevated in both groups, without significant differences. The clinical implication of elevated DPD activity in T1, T2, node negative or ER positive breast cancer compared to the respective normal breast tissue activity remains to be studied, because it is still unclear whether or not the tumoral DPD activity regulates the local concentration of 5-FU within the tumor. The amount of TS and DPD activity was not influenced by the oral administration of 5-FU for 2 weeks at the dose of 200 mg/day. PMID- 11057324 TI - [Antitumor activity of UFT and docetaxel on human breast carcinoma xenografts]. AB - The effect of fluorinated pyrimidine co-administered with docetaxel on transplantable human breast cancer strains MX-1 and R-27 was investigated using an in vitro succinic dehydrogenase inhibition (SDI) method and an in vivo nude mouse transplant method. The in vivo results showed that the combined use of 5 fluorouracil and docetaxel enhanced antitumor activity. No antitumor activity on MX-1 was observed in vivo in either the UFT alone or docetaxel alone group, whereas co-administration of the two drugs resulted in a tumor inhibition rate of 85.1% above the effective line. These results suggest the usefulness of 5 fluorouracil in combination with docetaxel in the treatment of solid tumors. PMID- 11057325 TI - [A noteworthy case of postoperative liver metastasis from gastric cancer which responded well to UFT therapy]. AB - A 66-year-old male underwent partial gastrectomy (resection of the pyloric side of the stomach) at another hospital on the basis of a diagnosis of gastric cancer (Fig. 1). Five months later, his CEA level began to rise. At that time, multiple liver metastases were detected by ultrasonography and CT scans. The patient received oral UFT therapy (400 mg/day) at our hospital. A reduction in CEA was observed 63 days after the start of this therapy. A judgment of CR (complete response) was made after 4 months of the therapy. At present, 2 years and 4 months after UFT was first administered, the patient shows no signs of tumor recurrence. This case is noteworthy since there has been no previous report of a case where UFT showed a high efficacy in treating liver metastasis after surgical resection of gastric cancer. PMID- 11057326 TI - [Two patients with liver metastasis from gastric cancer who responded remarkably to combined therapy of 5'-DFUR and PSK]. AB - Two patients with liver metastasis from gastric cancer who responded remarkably to a combined therapy of 5'-DFUR and PSK are reported. The first patient had liver metastases 8 months postoperatively. Her AFP level was 513 ng/ml and CEA was 30 ng/ml. After the combined therapy, all liver metastases showed a calcified change. Tumor markers of AFP and CEA decreased remarkably to the normal level within 3 weeks after the therapy. The patient had no relapse as of December 1999. The second patient with liver metastases was treated using the same combined therapy for 14 days preoperatively. The size of liver metastases had decreased remarkably by the time of the operation. A small metastasis of S8 (0.5 x 0.5 cm) was resected. No other liver metastasis was detected by intraoperative ultrasonography. The patient had no relapse as of February 2000. It is reported that PSK produces several cytokines which induce thymidine phosphorylase expression. The present report suggests that the upregulation of PyNPase might enhance the antitumor effect of 5'-DFUR. PMID- 11057327 TI - [A case of large-cell lung cancer responding remarkably to combination chemotherapy with vinorelbine, mitomycin C and carboplatin]. AB - The patient was a male who started to show symptoms at age 59. He was a smoker until age 40. In October 1998 he came to the hospital complaining of hemosputum and hoarseness. There was already swelling of the supraclavicular lymph nodes. Through lymph node aspiration cytology and bronchofiberscopy, large-cell carcinoma (T2N3M0, stage IIIB) was diagnosed. Chemotherapy with vindesine (VDS, 3 mg/m2), mitomycin C (MMC, 8 mg/m2) and carboplatin (CBDCA, 300 mg/m2) was conducted in three stages. Thanks to a partial response (PR) the patient was released in January 1999. However, in September 1999 he was readmitted when dysphagia, loss of body weight and dyspnea appeared. After bronchoscopy, chemotherapy combining vinorelbine (VNB, 25 mg/m2), (MMC, 8 mg/m2), CBDCA and the Calbert method calculated at AUC = 4.5 (AUC = area under the concentration-time curve) was completed in 4 stages. Upon PR and an abatement of symptoms he was released from the hospital. It is thought that treatment combining VNB is effective. PMID- 11057328 TI - [A case of docetaxel-resistant lung cancer effectively treated with chemotherapy of gemcitabine and vinorelbine]. AB - We report a NSCLC patient effectively treated with anti-cancer agents on an outpatient basis despite chemotherapy with carboplatin and docetaxel, he relapsed after achieving a transient response. Combination chemotherapy with gemcitabine and vinorelbine administered every three weeks in an outpatient clinic caused the lung cancer volume to reduce remarkably. This regimen was well tolerated and suitable for outpatients. Its usefulness as a second line chemotherapy in the treatment of NSCLC should be investigated. PMID- 11057329 TI - [Two bedridden patients with bone metastases from breast cancer effectively treated with pamidronate therapy]. AB - Case 1: A 43-year-old woman underwent mastectomy because of locally advanced breast cancer with multiple bone metastases. She was treated with CMF therapy but developed a compression fracture of a thoracic vertebra after 10 months and received pamidronate therapy. Pamidronate administration relived her back pain after 2 months and she was able to walk again after 3 months. However, she developed a resistance to the treatment, and then refused another treatment. She was found to have hypercalcemia 6 months later and received pamidronate again, but died 9 months after the treatment. Case 2: A 52-year-old woman underwent mastectomy because of breast cancer (T2) and was diagnosed as having multiple bone metastases 24 months after the operation. She could not turn over in bed due to progressing bone pain and received pamidronate therapy with CMF therapy at home 23 months after the diagnosis. After 2 months, pamidronate administration relieved her bone pain and she was free of pain after 4 months. After 5 months, X rays revealed that lytic lesions showed sclerosis, and the pamidronate therapy was assessed as producing a PR. Pamidronate therapy improved her quality of life and activities of daily living, and she continues to receive it this time as an outpatient. Pamidronate therapy is promising as an effective treatment for bedridden patients with bone metastasis from breast cancer. PMID- 11057330 TI - [Concurrent chemoradiotherapy using a single agent for unresectable stage III non small cell lung cancer]. PMID- 11057331 TI - [Dose escalation strategies of molecular target drugs]. AB - During the last decade, advances in cell biology and molecular biology have produced molecular targets for cancer treatment, and many molecular targeted drugs, or "cytostatic agents," have been clinically evaluated. Dose limiting toxicities are not always observed with these drugs and there is little relationship between drug dose and biological effect. Therefore, the traditional strategy used in the clinical development of cytotoxic drugs may not be appropriate for evaluation of these cytostatic agents. In order to determine the optimum dose for these agents, new clinical strategies must be developed on the basis of the following considerations: 1. the relationship between the effective plasma concentration in preclinical data and mean trough plasma levels in clinical trials, 2. changes in plasma concentration in the tumor or expression of molecular targets according to dose escalation, and 3. application of surrogate markers for biological activity. PMID- 11057332 TI - 60th Annual Congress of the Swiss Society of Orthopedics. Davos, 7-9 September 2000. Abstracts. PMID- 11057333 TI - 9th Annual Rotman Research Institute Conference. Traumatic Brain Injury: Diagnosis, Outcome, and Rehabilitation. March 24-26, 1999. Abstracts. PMID- 11057334 TI - Academy of Aphasia 2000. October 2000. Abstracts. PMID- 11057335 TI - Annual meeting and postgraduate course of the Cardiovascular and Interventional Radiological Society of Europe, and joint meeting with the European Society of Cardiac Radiology (ESCR) and joint meeting with the 3rd Basic Multidisciplinary Haemodialysis Access Course. Maastricht, The Netherlands, September 10-14, 2000. Abstracts. PMID- 11057336 TI - 10th North American ISSX meeting. October 24-28, 2000. Indianapolis, Indiana, USA. Abstracts. PMID- 11057337 TI - 35th Congress of the European Society for Surgical Research (ESSR). Malmo, Sweden, June 1-3, 2000. Abstracts. PMID- 11057339 TI - Proceedings of the Physiological Society scientific meeting held at University of Cambridge, 18-20 July 2000. Abstracts. PMID- 11057338 TI - Vessels and hormones 2000. Meeting of the National Society of Vascular Medicine and 11th European Conference on Clinical Hemorheology. 20-22 September 2000, Rouen, France. Abstracts. PMID- 11057340 TI - XVIII Annual scientific meeting of the British Blood Transfusion Society 2000. Nottingham, United Kingdom, 7-10 September 2000. Abstracts. PMID- 11057341 TI - Cytokines, eosinophils, lymphocytes, and ICAM. PMID- 11057342 TI - A comparison of the effects of olopatadine and ketotifen on model membranes. AB - Olopatadine is a human conjunctival mast cell stabilizer with antihistaminic activity. Ketotifen is an older molecule that possesses antihistaminic activity and is reported to have additional pharmacological properties. The interactions of these two compounds with model membranes (i.e., monolayers of 1-stearoyl-2 oleoyl-sn-glycerophosphocholine at the argon-buffer interface), and natural (i.e., erythrocyte) membranes were compared in an effort to understand the differences in their biological activities. Drug-lipid interaction with monolayers was determined by monitoring the surface pressure as a function of the drug concentration in the aqueous phase supporting the monolayer. Drug interaction with erythrocyte membranes was determined by monitoring changes in the permeability of the membranes to hemoglobin and 6-carboxyfluorescein as a function of drug concentration in the medium. Olopatadine and ketotifen are both intrinsically surface active and both interact with phospholipid monolayers. However, in both the presence and absence of lipid monolayers, the changes in surface pressure induced by olopatadine are lower than those caused by ketotifen. The effects of these two drugs on cell membranes were dramatically different. Exposure of bovine erythrocytes to increasing concentrations of ketotifen (1-10 mM) resulted in complete hemolysis of the cells, whereas olopatadine (1-10 mM) caused only minimal hemolysis (< 8%). Consistent results were obtained in experiments measuring the leakage of 6-carboxyfluorescein from erythrocyte ghosts as a more sensitive marker of membrane perturbation. Olopatadine treatment (0.1 10 mM) minimally perturbed the cell membrane while ketotifen (1-10 mM) caused a concentration dependent release of the fluorescent marker. These data demonstrate fundamental differences between the two drugs in their effects on cell membranes. Moreover, the differences are consistent with the surface activities of the two compounds measured in monolayers and with reported differences in their pharmacological activities. These findings offer an explanation for the biphasic non-specific cytotoxic effect of ketotifen on histamine release from mast cells and may account for the nonlytic mast cell stabilizing activity of olopatadine. PMID- 11057343 TI - A laboratory model to determine the uptake and release of olopatadine by soft contact lenses. PMID- 11057344 TI - Role of histamine in allergic conjunctivitis. AB - Histamine remains the main mediator released by both specific allergic and non specific mast cell activation. Histamine is the classic mediator of itching, flare and redness. The effects of histamine in allergic conjunctivitis are mediated by H1-receptor activation on blood vessels and nociceptive nerves. Histamine effects may be prolonged and exaggerated since a defect in the histaminase enzymes has been recently demonstrated in VKC. The effects of histamine on conjunctival tissues may be more complex than those manifested by the simple symptom of itching. In fact, proinflammatory effects of histamine on conjunctival epithelial cells and fibroblasts have been demonstrated. Preliminary results showed that the H1 antagonist, emedastine, reduces significantly cytokine production from histamine-stimulated fibroblasts. PMID- 11057345 TI - Immunology of uveitis and ocular allergy. PMID- 11057346 TI - The effects of systemic medication on ocular allergic disease. PMID- 11057347 TI - Ocular allergic disorders and dry eye disease: associations, diagnostic dilemmas, and management. PMID- 11057348 TI - Epidemiological study of 134 subjects with allergic conjunctivitis. AB - At times it is difficult to distinguish among the subtypes of chronic allergic conjunctivitis. A prospective study with 134 patients was carried out. Patients were diagnosed with vernal keratoconjunctivitis (VKC), atopic keratoconjunctivitis (AKC) or perennial allergic conjunctivitis (PAC). Demographic information was also collected including, age at symptom onset, sex, history of non-ocular allergy, and history of family allergy. Forty-six percent were diagnosed with VKC, 40% with AKC, 8% with PAC, and 6% were not specifically diagnosed. This distribution information as well as the demographic information collected demonstrates that there are patterns within each subgroup of chronic ocular allergy sufferers. These findings will be helpful in the diagnosing and proper classification of ocular allergic conditions. PMID- 11057349 TI - The early and late phase of the ocular allergic reaction. PMID- 11057350 TI - Safety and efficacy comparison of emedastine 0.05% ophthalmic solution compared to levocabastine 0.05% ophthalmic suspension in pediatric subjects with allergic conjunctivitis. Emadine Study Group. AB - PURPOSE: To determine the efficacy and tolerance of emedastine 0.05% ophthalmic solution compared to levocabastine 0.05% ophthalmic suspension in pediatric subjects. METHODS AND MATERIALS: In a randomized, double-masked, parallel controlled study, emedastine 0.05% ophthalmic solution BID was compared to levocabastine 0.05% ophthalmic suspension BID, for control of the signs and symptoms of allergic conjunctivitis in pediatric subjects ages 3-16. Subjects who met all inclusion and exclusion criteria received masked study medication with instructions to instill drops twice daily, in the morning and evening. A diary was completed by the parents four times daily for the first two and last two weeks of the study. Treatment lasted 42 days. Drug efficacy was assessed at the initial administration in the office at Day 0 and after 3, 7, 14, 30 and 42 days. RESULTS: Overall results showed both drugs have an effect and that emedastine was significantly superior (p < 0.05) to levocabastine for the relief of chemosis on Days 14, 30 and 42; of itching on follow-up Days 30 and 42 (p < 0.05); of redness on Days 30 and 42; for eyelid swelling on Days 14 and 30; and for physician's impression score on Days 7, 14, 30 and 42. CONCLUSION: These results confirm previous preclinical and clinical data on the potent and long acting efficacy of this promising new ophthalmic anti-allergic drug, emedastine in pediatric subjects. PMID- 11057351 TI - An efficacy and tolerance comparison of emedastine difumarate 0.05% and levocabastine hydrochloride 0.05%: reducing chemosis and eyelid swelling in subjects with seasonal allergic conjunctivitis. Emadine Study Group. AB - PURPOSE: To compare emedastine ophthalmic solution 0.05% BID to levocabastine ophthalmic suspension 0.05% BID in reducing chemosis, eyelid swelling and other signs and symptoms in subjects with seasonal allergic conjunctivitis. METHODS: In a randomized, double-masked, parallel controlled study, emedastine ophthalmic solution 0.05% BID was compared to levocabastine ophthalmic suspension 0.05% BID for control of chemosis, eyelid swelling and other parameters in the environmental allergy study model. RESULTS: At Days 7, 14, 30 and 42, emedastine was significantly better than levocabastine at controlling chemosis and eyelid swelling (p < 0.05). A statistical trend was seen at Day 3 (0.05 < p < 0.10). Results were clinically relevant at Days 30 and 42. Emedastine was also significantly better at reducing redness and itching at Days 7, 14, 30 and 42 (p < 0.05). CONCLUSION: Emedastine is more efficacious than levocabastine in reducing chemosis, eyelid swelling and other efficacy variables associated with seasonal allergic conjunctivitis. PMID- 11057352 TI - Comparative study of clinical efficacy and tolerance in seasonal allergic conjunctivitis management with 0.1% olopatadine hydrochloride versus 0.05% ketotifen fumarate. AB - OBJECTIVE: To compare the clinical efficacy and tolerance of 0.1% olopatadine hydrochloride (OHC) versus 0.05% ketotifen fumarate (KF) in the management of allergic conjunctivitis. MATERIALS AND METHODS: Eighty adult patients with a history of allergy (allergic conjunctivitis, hay fever, asthmatic bronchitis and dermatitis) that were showing allergic conjunctivitis signs and symptoms (itching, hyperemia, mucous discharge and tearing) at the time of inclusion in this study were evaluated. Patients were divided in two groups, A and B. Group A patients were treated with OHC and group B patients were treated with KF. Both groups received one drop in the affected eye every 12 hrs. The start time of this study was the first patient visit, in which the medication was instilled for the first time. Both groups of patients were evaluated 30 min, 48 hr., 7 days and 14 days later. Local tolerance of each medication was evaluated. RESULTS: Clinical improvement of the signs and symptoms of allergic conjunctivitis occurred in 42.5% to 62.5% of the patients in Group A when assessed 30 min following the first topical ocular dose of olopatadine. However, mucous discharge was not affected. Forty-eight (48) hrs. after the first instillation, improvements in 57.5% to 75% of the patients were shown in every evaluated parameter. After 7 days of treatment, positive clinical results were observed in 80% to 87.5% of the treated patients. Except for the patients that were dismissed from the study before the seventh day of treatment due to the absence of therapeutic response (4/40), all patients satisfactorily completed the therapeutic plan by the seventh day. No intolerance reactions were observed in patients of this group. In Group B patients (KF), clinical improvement of the signs and symptoms measured in the study was shown in 20.0% to 47.5% 30 min after instillation. As observed with olopatadine, no improvement in the number of patients showing mucous discharge was noted at the 30-min time point. At 48 hr. after the first instillation, 27.5% to 48% of patients showed improvement in every evaluated parameter. After 7 days of treatment, improvement was observed in 60% to 75% of patients. On Day 14, positive responses were observed in 67.5% to 75% of patients. Seventeen and one half percent of the patients were dismissed from the study before the seventh day of treatment due to the absence of a therapeutic response. Approximately 23% of the patients had mild reactions of intolerance (stinging) which was not a cause to discontinue the treatment. CONCLUSION: Olopatadine hydrochloride controlled allergic conjunctivitis symptoms and signs more rapidly and to a greater extent than ketotifen fumarate. Fewer cases of treatment failure were noted with OHC, and no local intolerance reactions were observed, while KF triggered mild reactions (stinging) in 23% of patients. PMID- 11057353 TI - Allergic conjunctivitis and contact lenses: experience with olopatadine hydrochloride 0.1% therapy. AB - OBJECTIVE: The efficacy of Patanol, a topically applied anti-allergic agent, was evaluated in a group of patients with allergic conjunctivitis due to contact lens wear (GROUP I) and a group comprised of seasonal allergic conjunctivitis patients, vernal conjunctivitis patients and atopic keratoconjunctivitis patients (GROUP II). METHODS: One drop of Patanol was administered to each eye twice daily. Signs and symptoms were assessed 7, 14, and 28 days after initiation of drug therapy. RESULTS: Itching/burning, tearing, hyperemia and papillary reaction were reduced to scores of 0/1 (absent/mild) in 85%, 90%, 81% and 62%, respectively, of GROUP I patients at Day 28. The allergic conditions in GROUP II patients also improved with Patanol treatment. Itching/burning, tearing, hyperemia and papillary reactions were absent/mild in 60%, 76%, 96% and 90%, respectively, of these patients at Day 28. CONCLUSION: Patanol treatment effectively and rapidly alleviated the signs and symptoms of allergic conjunctivitis due to contact lens wear as well as vernal conjunctivitis, atopic keratoconjunctivitis and the common seasonal allergic conjunctivitis. Patanol allowed allergic patients to be more comfortable while permitting them to continue using contact lenses. PMID- 11057354 TI - An evaluation of onset and duration of action of patanol (olopatadine hydrochloride ophthalmic solution 0.1%) compared to Claritin (loratadine 10 mg) tablets in acute allergic conjunctivitis in the conjunctival allergen challenge model. AB - PURPOSE: To compare the clinical efficacy of Patanol (olopatadine hydrochloride ophthalmic solution 0.1%) to Claritin (loratadine 10 mg) tablets, in the conjunctival allergen challenge model. METHODS: This was a randomized, double masked, single center, contralateral controlled, antigen challenge model study. The concentration of allergen that elicited a positive response was determined at Visits 1 and 2 (itching > or = 2 and redness > or = 2 OU). At Visit 3, 29 subjects were randomized into two groups. Fifteen subjects received Claritin tablet and Patanol ophthalmic solution 0.1% in one eye and placebo in the contralateral eye. Fourteen subjects received placebo tablet and Patanol in one eye and placebo in the contralateral eye. One hour after drug administration, subjects were challenged with the antigen that elicited a positive response. At 3, 7, and 10 minutes, itching was subjectively evaluated. At Visit 4, the same procedure was followed as in Visit 3, but antigen challenge occurred 8 hours after drug instillation. RESULTS: Results were analyzed by eye. Eyes treated with Patanol (concomitant with placebo tablet) had significantly lower ocular itching scores when compared to eyes treated with placebo (concomitant with Claritin) at 3, 7 and 10 minutes in the onset of action evaluation (p < 0.05). Eyes treated with Patanol (concomitant with placebo tablet) had significantly lower ocular itching scores at 7 minutes and there was a statistical trend (0.05 < p < 0.1) at 10 minutes in duration of action evaluation. CONCLUSIONS: Patanol therapy was significantly more efficacious than Claritin in reducing ocular itching related to allergic conjunctivitis. PMID- 11057355 TI - A forced choice comfort study of olopatadine hydrochloride 0.1% versus ketotifen fumarate 0.05%. AB - PURPOSE: To compare the ocular comfort of two ophthalmic anti-allergic agents: olopatadine hydrochloride 0.1% and ketotifen fumarate 0.05%. SUBJECTS AND METHODS: In a double-masked, multi-centered, randomized trial, 80 subjects were asked to make a 'forced choice' based on ocular comfort between one drop of olopatadine hydrochloride 0.1% instilled in one eye and one drop of ketotifen fumarate 0.05% instilled in the contralateral eye. At one site, the incidence of adverse reactions was also reported. RESULTS: All subjects (100%) selected olopatadine as the more comfortable formulation. One site (n = 35) reported a 49% incidence of moderate burning and a 49% incidence of mild burning after ketotifen instillation. One subject (2% of population) at this site experienced no ocular discomfort with ketotifen. There were no reports of discomfort associated with olopatadine instillation. CONCLUSION: Olopatadine is a more comfortable ophthalmic preparation than ketotifen. PMID- 11057356 TI - Allergic rhinoconjunctivitis--an overview. PMID- 11057357 TI - Ocular allergies: association with immune dermatitis. AB - PURPOSE: To describe the type of skin involvement that patients with ocular allergies are prone to suffer due to the atopic association of the condition. METHODS: Review of the published literature and retrospective data from our patient series. RESULTS: Patients with ocular allergies may have skin affected with one or several combinations of the following dermatologic problems: contact dermatitis, urticaria/angioedema, and atopic dermatitis, with secondary infections as a possible consequence of this later disorder. Other rare skin disorders have been occasionally reported in association with ocular atopy. CONCLUSION: Patients with ocular allergies may have a spectrum of 'allergic' skin problems, the most severe of which is atopic dermatitis. Ocular surface involvement in atopic dermatitis should be diagnosed as atopic keratoconjunctivitis, a sight-threatening disorder, until proven otherwise. PMID- 11057358 TI - Skin tests and cutaneous anergy in children with ocular allergy. AB - PURPOSE: To evaluate skin tests with clinical ocular allergy and to compare cutaneous hypersensitivity and seasonal allergic conjunctivitis in children. CASE REPORT: A 10-year-old girl presented a typical seasonal allergic conjunctivitis, never treated and the onset occurred one year before. She was skin tested for the allergy. Complete blood tests were also performed. An ocular treatment was given: olopatadine over 45 days. RESULTS: She showed a complete cutaneous anergy including histamine and codeine controls. Radioallergosorbent tests (RAST) found numerous great positivity to different allergens. Within 30 days under ocular treatment, she recovered completely from symptoms and clinical signs. CONCLUSION: Clinical typical seasonal ocular allergy in children with cutaneous anergy should be explored further or at least treated. Cutaneous hypersensitivity should be evaluated more precisely with a larger study comparing results in general allergy and in ocular allergy where the anergy seems to be more frequent. PMID- 11057359 TI - Immune-mediated corneal melting disease. AB - Corneal melting disease is, as illustrated, a very severe problem for patients. Correct diagnosis and aggressive management is critical. Help from general physicians and rheumatologists as required is essential, given that the medications used have significant toxic effects. PMID- 11057360 TI - Area under the inhibitory curve and a pneumonia scoring system for predicting outcomes of vancomycin therapy for respiratory infections by Staphylococcus aureus. AB - Treatment factors predictive of clinical and microbiological outcomes and the relationship between a pneumonia scoring system and clinical outcomes in vancomycin-treated patients with a Staphylococcus aureus-associated lower respiratory-tract infection (LRTI) were studied. A computer database review identified patients for whom S. aureus was isolated from a respiratory-tract specimen between January 1 and December 31, 1998, and who had antimicrobials ordered within 72 hours of isolation of that organism. Through further review of individual patient charts, this group was restricted to those treated with vancomycin for a documented S. aureus-associated LRTI. Classification-and regression-tree (CART) modeling was performed to determine which clinical variables were correlated with clinical outcomes and microbiological outcomes. Median changes in clinical pneumonia scores from baseline in two patient groups (those with clinical success and those with clinical failure) were compared. Seventy patients met the study criteria. CART modeling found that both outcomes were associated with area under the inhibitory curve (AUIC). The pneumonia scoring system was predictive of eventual clinical success as early as day 3 of treatment; having at least a 4-point decrease in the pneumonia score by day 3 was correlated with an 87% clinical success rate. Both AUIC and a pneumonia scoring system were useful for predicting clinical and microbiological outcomes of vancomycin therapy in patients with LRTIs caused by S. aureus. PMID- 11057361 TI - Economic justification of antimicrobial management programs: implications of antimicrobial resistance. AB - The relationship between the problem of antimicrobial resistance and efforts to control antimicrobial costs is explored. Antimicrobial drug management typically centers around controlling costs and controlling antimicrobial resistance. Selection of therapeutic alternatives without adherence to a well-developed program or without a rationale based on data from the medical literature may promote antimicrobial resistance. Attempts to select alternatives can produce cost shifting rather than cost containment. The annual cost associated with antimicrobial resistance in the United States is estimated to be as high as $47 billion. In one study, patients with bacteremia caused by methicillin-resistant Staphylococcus aureus had an average length of stay 2.7 days longer than patients with susceptible strains and a mean cost of care that was $3500 higher. Infection control is one of the most important duties of health care practitioners. Given today's prevailing reimbursement structure, hospitals with high rates of nosocomial and resistant infections are likely to lose money. A basic problem with the current approach to controlling resistance is that the two most common strategies, highly restrictive formularies and drug cycling, work in opposition. Antimicrobial management programs should be directed at ensuring the most appropriate use of antimicrobials rather than focusing on limiting choices. PMID- 11057362 TI - Benchmarking in health-system pharmacy: current research and practical applications. AB - The application of benchmarking techniques to hospital pharmacy practice is discussed. Benchmarking is a process designed to discover best practices through a comparison of various competing methods aimed at achieving a particular goal. Benchmarking antimicrobial drug utilization and rates of bacterial resistance through comparison with a multitude of similar hospitals can be used by an institution both to identify potential problem areas in its pharmacy practice and to aid in establishing appropriate and attainable goals. The effectiveness of various activities targeted at reducing appropriate drug use can also be benchmarked. In 1993, the Benchmarking Program was established at Millard Fillmore Hospital. This program consists of a network of hospital pharmacists who supply data on antimicrobial use, antimicrobial management activities, and rates of antimicrobial resistance. The program was designed both to serve hospital pharmacies in optimizing antimicrobial management and to create a national database for evaluating relationships among antimicrobial use, management, and resistance. Hospitals participating in the Benchmarking Program receive an annual report that allows them to compare themselves with peer groups and with best performing "benchmark hospitals." All data from U.S. hospitals contained in the Benchmarking Program database are pooled and analyzed to identify meaningful trends. However, information gained from the institutionwide data must be supplemented by studies at the patient level. Benchmarking antimicrobial drug use in an institutional setting can identify successes as well as potential problem areas in pharmacy practice and aid in establishing appropriate and attainable goals. PMID- 11057363 TI - Benchmarking in health-system pharmacy: experience at Glens Falls Hospital. AB - The experience of Glens Falls Hospital (GFH) with the Benchmarking Program coordinated by The Clinical Pharmacokinetics Laboratory at Millard Fillmore Hospital is described. GFH, a community hospital in upstate New York with 442 licensed beds, serves a patient population drawn from a five-county area. In 1998, GFH developed a multidisciplinary pharmacy task force charged with generating strategic initiatives for curtailing drug costs. Proposals for various programs, including specific goals, were developed with the aid of trends reported in the individual hospital reports provided by the Benchmarking Program. The data obtained through these reports helped GFH's pharmacy staff establish goals for the appropriate use of many agents, including vancomycin, and allowed GFH to assess the impact of its initiatives through comparisons with similar hospitals across the United States. In 1995, vancomycin expenditures at GFH substantially exceeded those for benchmark hospitals of similar types, but, with the implementation of new vancomycin guidelines aimed at reducing inappropriate use in 1996, the difference was eliminated. The individual hospital reports generated by the Benchmarking Program provide valuable data that can be used to assess pharmacy performance, set drug expenditure goals, and design programs to reduce antimicrobial resistance. PMID- 11057364 TI - Benchmarking in health-system pharmacy: experience at MeritCare Medical Center. AB - The experience of MeritCare Medical Center (MMC) with the Benchmarking Program coordinated by The Clinical Pharmacokinetics Laboratory at Millard Fillmore Hospital is described. MMC is a community-based teaching institution in Fargo, North Dakota, that serves patients in North Dakota, South Dakota, and Minnesota. MeritCare began participating in the Benchmarking Program in 1997. Data from the individual hospital report raised concern about the high cost of antimicrobials at MMC relative to peer-group institutions. The staff conducted an evaluation of antimicrobial prophylaxis for noncardiovascular surgery, concluded that cefazolin use was suboptimal, and attempted to encourage more cost-effective utilization. MMC's participation in the Benchmarking Program also prompted more appropriate use of various other antimicrobial agents, including i.v. and oral ciprofloxacin. An i.v.-to-oral switching program was begun for various agents. Preliminary analysis after 15 months demonstrated direct cost savings for drug acquisition of $60,000 to $80,000 per year and a reduced length of stay. Initiatives undertaken by MeritCare on the basis of data obtained through the Benchmarking Program resulted in substantial estimated savings in drug acquisition costs. PMID- 11057365 TI - Using benchmarking data to evaluate and support pharmacy programs in health systems. AB - The use of benchmarking data to evaluate and support pharmacy programs in health systems is discussed. Benchmarking is a method of comparing the outcomes of health care products, services, and practices at an institution against those of competitors in order to learn what might be improved. Benchmarking programs can provide valuable feedback about both positive and negative outcomes. However, it is imperative to avoid inappropriate comparisons and inappropriate assessments. Ideal services for benchmarking are those that have a good likelihood of improving patient care and other outcomes. Successful benchmarking requires sound and thorough data, which is why as many health systems as possible should participate in a benchmarking program. The National Committee for Quality Assurance has formulated guidelines that should enable health systems to develop an information framework that will improve their ability to collect and use data for benchmarking. Standardization in collecting and submitting information is important because it enables institutions to share data easily. Benchmarking can help health-system pharmacists understand the value and outcomes of efforts by their colleagues at other institutions. In addition, benchmarking can help convince health-system administrators of the value of pharmaceutical services in terms of patient care and the bottom line. Benchmarking provides a means of evaluating and supporting the development of pharmacy programs that improve care and save money. PMID- 11057366 TI - [Antibiotics and problems of pharmacoenzymology]. PMID- 11057367 TI - [Experimental resistance of Vibrio cholerae el tor to nalidixic acid and fluoroquinolones]. AB - It was shown that sensitivity of Vibrio cholerae eltor P-5879 to tetracycline, levomycetin, furazolidone, trimethoprim/sulfamethoxazole, aminoglycosides, beta lactams, rifampicin, quinolones in vitro correlated with drugs efficacy in the treatment of experimental cholera of albino mice. Mutants of V. cholerae eltor P 5879 Nalr resistant to nalidixic acid (MIC 160-200 mg/l) formed with frequency 10(-9)-110(-8) had no cross resistance to fluoroquinolones. But the efficacy of ofloxacin, lomefloxacin, norfloxacin against these mutants in vivo reduced, though it was not changed in vitro. Mutants of V. cholerae eltor P-5879 resistant to fluoroquinolones and selected after culturing in the presence of the drugs had cross resistance to all quinolones studied. Infection caused by Cpfr mutant could not be treated with nalidixic acid and fluoroquinolones, therapeutic efficacy of rifampicin and beta-lactams, also reduced though sensitivity in vitro was not changed. The results of investigation proves the necessity of quinolones use for cholerae treatment as it is recommended for other severe enteric infections. PMID- 11057368 TI - [Spectrum of pneumotropic pathogens in pediatric patients with acute bronchitis and pneumonia]. AB - One hundred eighty nine children with acute bronchopulmonary infectious pathological processes were examined bacteriologically and serologically for typical pneumotropic pathogens, 47 of them being as well examined for atypical organisms. Microbial associations mainly with Mycoplasma and Pneumocystis and to a lesser extent with Chlamydia were isolated from the majority of the children. Reactivation of the cytomegalovirus infection was observed in 25 per cent of the children. Pneumonia and bronchitis due to Mycoplasma pneumoniae either as a monoagent or in associations were mainly stated in children over 7 years of age. No significant changes between the indices of the infection due to a definite organism and the active progression of the infectious process of the same etiology were revealed, though in the cases of chlamydiosis the changes reached almost 10 per cent. In cases of acute bronchitis and pneumonia the chlamydial or cytomegalovirus infection could be assumed to be of the persisting nature, mainly acute in cases of pneumococcal infection, mixed in cases of hemophilic or pneumocystic infection, primary contamination with a tendency to prolonged in cases of mycoplasmic infection. The findings of the examination and the clinical and anamnestic data showed that the clinical picture of acute pneumonia had specific features associated with the supposed etiological agents, still it could change under the action of associations of pneumotropic pathogens. PMID- 11057369 TI - [Preventive use of antibiotics in clinical therapy]. PMID- 11057370 TI - [Functions of gastrointestinal microflora and its postoperative disorders]. PMID- 11057371 TI - [The new technologies in chemotherapy of tuberculosis]. PMID- 11057372 TI - [The role of fluoroquinolones in treatment of pneumonia]. PMID- 11057373 TI - Phallometric testing with sexual offenders: limits to its value. AB - In many settings phallometric evaluations of sexual arousal are routinely conducted with sexual offenders and these evaluation procedures also serve as research instruments. There are, however, problems with the psychometric bases of these assessments, and studies reporting their use have so many idiosyncratic features that comparisons are of dubious value. Evidence concerning the reliability and criterion validity of phallometric testing leaves a lot to be desired, although the research has suggested an important but limited value in predicting subsequent recidivism. Suggestions are made for further research and for the clinical use of phallometric assessments within more comprehensive evaluations of sexual offenders. PMID- 11057374 TI - Issues in the assessment and conceptualization of personality disorders. AB - This article reviews several current issues associated with the definition and assessment of personality disorders (PDs) as defined in the third and fourth editions of the Diagnostic and Statistical Manual of Mental Disorders (DSM). Specifically reviewed are issues associated with classification, PD conceptualizations, and the assessment of these disorders. DSM PD categories are also reviewed in terms of their psychometric properties. A review of the PD assessment literature suggests that DSM conceptualizations and definitions of PDs are problematic at both conceptual and quantitative levels. This article concludes with suggestions for possible alternative approaches to and modifications of DSM PD assessment. PMID- 11057375 TI - Apparent symptom overreporting in combat veterans evaluated for PTSD. AB - Psychometric studies have consistently shown that combat veterans evaluated for posttraumatic stress disorder (PTSD) appear to overreport psychopathology as exhibited by (a) extreme and diffuse levels of psychopathology across instruments measuring different domains of mental illness, and (b) extreme elevations on the validity scale of the MMPI-MMPI-2, in a "fake-bad" direction. The phenomenon of this ubiquitous presentational style is not well understood at present. In this review we describe and delineate the assessment problem posed by this apparent symptom overreporting, and we review the literature regarding several potential explanatory factors. Finally, we address conceptual and practical issues relevant to reaching a better understanding of the phenomenon, and ultimately the clinical syndrome of combat-related PTSD, in both research and clinical settings. PMID- 11057376 TI - Childhood emotional abuse and eating psychopathology. AB - The potential role of childhood emotional abuse (CEA) in the etiology and maintenance of eating psychopathology is reviewed. While childhood sexual and physical abuse have been hypothesized as risk factors in multifactorial models of eating disorders, a role for CEA has only recently been considered. Initial findings demonstrate a phenomenological link between CEA and eating psychopathology, and suggest that this association might be different to the links for other forms of trauma (i.e., CEA may have a relationship with a broader range of eating symptoms than sexual and physical abuse). However, the psychological processes that might account for such a link are not yet well understood. Potential cognitive and affective mediators are considered, with a particular emphasis upon low self-esteem and anxiety. A model is proposed, to act as a framework for further research into this field. The clinical implications of the research to date and of the proposed model are discussed. PMID- 11057377 TI - Racial, ethnic, and cultural factors of childhood sexual abuse: a selected review of the literature. AB - Considerable research on childhood sexual abuse exists; however, few studies have examined the role of race, ethnicity, and culture in such abuse. Past investigations have focused almost exclusively on several ethnic groups and races, namely whites, blacks, and Hispanics, without much delineation of cultural subgroup differences. Additionally, much of the existing research is conflicting and lacks consistency in the use of ethnic and racial terms and identification of cultural variations among the major groups. This paper will summarize and analyze the literature on childhood sexual abuse in relation to racial, ethnic, cultural, and other factors relevant to African American, Hispanic, and Asian populations. A critique of the current weaknesses in the literature, including contradictions and recommendations for future research are also presented. PMID- 11057378 TI - Cardiac rehabilitation: a review of current developments. AB - An overview is given of the current status of cardiac rehabilitation and its effects on morbidity and mortality. While there is an emphasis in most current programs upon physical exercise as an important autonomous risk factor for Coronary Heart Disease (CHD), there is at the same time a tendency in cardiac rehabilitation to go beyond mere physical exercise towards adding more multimodal psychoeducational modules in rehabilitation programs; those approaches are aimed at educating the patient about a less risky and healthier way of life. Such psycho-education is more and more aimed at the "toxic" aspects of negative emotions. The in-between classic Type A Behavior Pattern (TABP) might, in general, be less powerful in predicting later CHD morbidity or mortality than some specific emotional components of TABP, such as anger and hostility. The literature is reviewed as to risk factors and CHD and the role of negative affectivity in development and or maintenance of CHD. Approaches for modification are discussed against the background of their effectivity in cardiac rehabilitation. The recent Dutch guidelines, issued by the Dutch Heart Foundation, appear to incorporate many of the elements mentioned in the research literature on cardiac rehabilitation. On a scientific level they form an excellent audit to evaluate and to contour efficiently the until-now very heterogeneous field of cardiac rehabilitation. PMID- 11057379 TI - Tough priorities. PMID- 11057381 TI - Biotechnology and monstrosity. Why we should pay attention to the "yuk factor". PMID- 11057380 TI - Intimations of immortality. PMID- 11057382 TI - On the sanctity of nature. PMID- 11057383 TI - The million dollar question. PMID- 11057385 TI - On ova commerce. PMID- 11057384 TI - Managed care, conflicts of interest, and quality. PMID- 11057386 TI - A sociological account of the growth of principlism. PMID- 11057387 TI - The traditional African perception of a person. Some implications for bioethics. PMID- 11057388 TI - Vaccinations against bad habits. PMID- 11057389 TI - [New aspects on the pathogenesis of bullous pemphigoid]. AB - Bullous pemphigoid (BP) is a subepidermal blistering autoimmune disease of the elderly. Autoantibodies are directed against two hemidesmosomal proteins, designated BP180 and BP230. While BP230 localizes intracellularly and associates with the hemidesmosomal plaque, BP180 is a transmembrane glycoprotein with an extracellular domain consisting of approximately 1000 amino acids. The non collagenous (NC) 16A domain, that encompasses 76 amino acids and localizes directly adjacent to the transmembrane region, has been identified as an immunodominant region of the BP180 ectodomain. In the majority of BP sera, circulating antibodies to BP180 NC16A are detected; their serum levels correlate with disease activity. Neonatal mice that are injected with rabbit anti-murine BP180 antibodies develop a BP-like subepidermal blistering disease demonstrating the biological importance of antibodies to BP180. The pathogenically relevant site on murine BP180 corresponds to a stretch of the NC16A domain on human BP180. In contrast to pemphigus, in BP, autoantibodies alone are not sufficient to induce blisters. In addition, complement activation, the infiltration of inflammatory cells and the release of proteases and various inflammatory mediators, including cytokines, are essential for lesion formation. In this review, we give an up-date on the pathogenesis of BP. PMID- 11057390 TI - [Immunosuppressive macrolides and their use in dermatology]. AB - The immunosuppressive macrolides are a novel class of antiinflammatory substances, which could supplant glucocorticosteroids for the topical treatment of some chronic inflammatory skin diseases. Cyclosporine A (CyA), well known from transplantation medicine for years, is licensed in Germany for oral treatment of psoriasis and atopic dermatitis but is not suitable for topical therapy. Tacrolimus (FK506) penetrates the inflamed epidermis and is regarded as the key immunosuppressive macrolide. Clinical trials have demonstrated the efficacy of FK506 ointment for atopic dermatitis, many case reports have been published regarding other inflammatory skin diseases. The ascomycin derivative ASM 981 has many of the properties of FK506 but less data is available at present. Sirolimus (Rapamycin) is structurally related to FK506 but has other biological effects since its molecular actions involve different biochemical pathways. A review of the biochemical and cellular properties, mode of action, therapeutic efficacy and unwanted side effects, as well as data from clinical trials and status of licensing, is given for the respective drugs. PMID- 11057391 TI - [Reduction of colonization of new mattresses with bacteria, moulds and house dust mites by complete mattress covers]. AB - BACKGROUND AND OBJECTIVE: We investigated if the colonisation of new mattresses with house dust mites, bacteria, and fungi could be reduced by using synthetic mattress covers as compared to common cotton covers. PATIENTS/METHODS: 84 healthy volunteers were assigned to two groups. Group A (n = 43) received the cotton covers, group B (n = 41) the synthetic covers which were made of a polyester microfaser with a polyurethane surface layer (Pro-Tex, Germed, Schwarzenbek, Germany). RESULTS: The mite antigen concentration after six months was significantly lower in group B. Three months after the start of the study counts of bacteria and moulds were significantly higher in group A compared to group B. CONCLUSIONS: It can be recommended that patients suffering from an allergy to mites or moulds may reduce their domestic allergen exposure by using the synthetic mattress covers tested in this study. Since cotton covers are very likely to become colonised by bacteria and moulds, they must be cleaned periodically (at least every 2nd-3d month). PMID- 11057392 TI - [Lipoma of the forehead]. AB - BACKGROUND AND OBJECTIVE: Lipomas should be included in the differential diagnosis of tumors located on the forehead. They require a different operative plan. PATIENTS/METHODS: Lipomas of the forehead were extirpated in ten patients. Dermatohistopathologic investigations confirmed the diagnosis lipoma in each case and exactly identified the anatomic layers surrounding the tumor. RESULTS: Histologic investigations revealed the localization of the lipoma in the submuscular layer in eight patients and in the subgaleal layer in two patients. CONCLUSIONS: Lipomas of the forehead should be differentiated into submuscular and subgaleal lipomas depending on the exact anatomic localization below the frontalis muscle or underneath the galea aponeurotica. This classification is simple and aids in operative planning. PMID- 11057393 TI - [Antibiotic susceptibility of Neisseria gonorrhoeae isolates in Berlin]. AB - BACKGROUND AND OBJECTIVE: Because of the increasing resistance of Neisseria gonorrhoeae, we studied the actual resistance of isolates in Berlin. PATIENTS/METHODS: 85 Neisseria gonorrhoeae isolates were collected between 1995 and 1997. Susceptibility testing was performed for penicillin G, tetracycline, spectinomycin, ceftriaxone, ciprofloxacin and azithromycin by agar dilution. RESULTS: 18.8% isolates were resistant or intermediately resistant to penicillin G (including 6 PPNG). 12.9% isolates were resistant, 43.5% intermediately resistant to tetracycline. One strain was resistant against ciprofloxacin, 4 isolates showed increased MIC values (0.06-0.5 mg/l), whereas 78 isolates were fully susceptible (< 0.007 mg/l). All isolates were susceptible to spectinomycin, ceftriaxone, and azithromycin. CONCLUSIONS: Penicillin G and tetracycline should be given only in cases of proven sensibility. Resistance against ciprofloxacin may occur, especially in isolates acquired in south-east Asia. Ceftriaxone, spectinomycin and azithromycin were active against all isolates. The actual resistance situation should be monitored. PMID- 11057394 TI - [Clinical aspects and immunopathology in 48 patients with pemphigus]. AB - BACKGROUND AND OBJECTIVE: Pemphigus is a rare intraepidermal autoimmune bullous disease. Two major variants, pemphigus vulgaris (PV) and pemphigus foliaceus (PF), are distinguished. The aim of this study was to document the clinical and immunopathological findings in all pemphigus patients who were diagnosed in the Department of Dermatology at the University of Wurzburg over the past 10 years. PATIENTS/METHODS: Based on a retrospective study, clinical and immunopathological findings in 48 patients with pemphigus were recorded. All patients had positive findings by direct and/or indirect immunofluorescence microscopy. RESULTS: Between January 1989 and August 1998, 48 patients were diagnosed with pemphigus at our institution; 31 patients had PV and 17 PF. The average age (+/- standard deviation) of PV patients was 55 (+/- 17) and of PF patients 60 (+/- 12) years. All PV patients showed involvement of mucous membranes and in 65% of cases, the skin was also involved. In contrast, PF patients had involvement only of the skin. By direct immunofluorescence microscopy, intercellular deposits of IgG and C3 were detected in 89% and 78% of PV cases, respectively. In PF, intercellular deposits of IgG were found in 94% and of C3 in 75% of cases. By indirect immunofluorescence microscopy on monkey esophagus, 94% of the PV and 88% of the PF patients revealed circulating serum antibodies. In 30 patients, we characterized the immune response by ELISA using recombinant desmoglein 1 and 3. All PF sera showed autoantibodies against desmoglein 1 and all PV sera against desmoglein 3. In PV with both mucous membrane and skin involvement, antibodies to both desmoglein 3 and 1 were detected. CONCLUSIONS: Our results confirm the correlation of the autoantibody profile with the clinical phenotype of pemphigus. PMID- 11057395 TI - [Ibuprofen-induced, transient ANA- and anti-histone positive leukocytoclastic vasculitis]. AB - Drug-induced leukocytoclastic vasculitis is clinically characterised by inflammation of small vessels and skin alterations, typically palpable purpura. Often the vasculitis is accompanied by fever, myalgia and malaise. All ages and sexes are equally affected. Besides the mentioned clinical features it may also present as macules, papules, vesicules, bullae, subcutaneous nodules or intermittent or chronic urticarial eruptions. Other organ systems may be affected. The patient's history is helpful in identifying the etiologic agent. If a drug is suspected as etiologic agent, a positive lymphocyte transformation test (LTT) supports the hypothesis. A 55 year old female patient developed an ANA and anti-histone antibody positive leukocytoclastic vasculitis with probable concomittant renal involvement after three weeks treatment with ibuprofen. Three months after discontinuation of the medication, the patient is without complaints and has a normal urine sedimentation and normalized immuno-autoantibodies. PMID- 11057396 TI - [Dorsal mucoid cyst--ganglion-like pseudocyst of the joint space]. AB - A myxoid cyst on the dorsum of the left index finger is presented. A connection of this pseudocyst to the underlying joint was shown by means of magnetic resonance imaging. Surgically the connecting tract to the distal interphalangeal joint was easily demonstrated and histologically a ductal structure was focally seen by means of serial sections. These structures also suggest such a connection. These findings confirm the view that this pseudocyst can be interpreted as a ganglion. Therapeutically complete excision with careful tying off of the channel to the joint appears to be the best method to avoid recurrence. The injection of sclerosing agents, a conservative treatment modality proposed by some authors, may be problematic in the light of the pathogenesis discussed here; however damage to the finger joint has not been so far observed. PMID- 11057397 TI - [Acrocephalosyndactyly I (Apert syndrome)]. AB - A fourteen years old girl showed the classic signs of acrocephalosyndactyly I: dysostosis craniofacialis with hypertelorism, exophthalmus, strabism, amblyopia and cleft palate as well as syndactyly of the fingers and toes. The feet showed on both side a 6 cm long horny band. Since the twelfth year of life, she had suffered from papulo-pustular acne with many comedomes. Her menstruation started one year later. Intellectual development was normal. At time of her birth, her father was 54 years old, and her mother 36 years old. Two elder siblings are healthy. The inheritance of acrocephalosyndactyly I is usually autosomal dominant, but sporadic cases are frequent. PMID- 11057398 TI - [Hypomelanosis Ito in translocation trisomy 9/mosaicism (46,XX/46,XX,t(9;9)(p24;p24)). Spontaneous remission in childhood]. AB - A 4 5/12 years old girl with hypomelanosis of Ito (HI) presented on the 3rd day of life with hypopigmented streaks and whorls following the lines of Blaschko on the back, the arms and the legs. In addition, patchy depigmented areas were present on the trunk. Extracutaneous manifestations included dystopia of the right kidney, atrial septal defect, persistent ductus arteriosus, hearing impairment, EEG abnormalities, and asymmetric dilatation of the ventricle system and a vermal atrophy as documented in the MRT of the brain. Cytogenetic analysis showed a mosaic 46,XX/46,XX,t(9;9)(p24;p24) present in the lymphocytes and skin fibroblasts. The mother's karyotype in her lymphocytes was normal. At reexamination at the age of 4 5/12 years the girl was retarded. In particular, the speech development was severely delayed. Interestingly we found only very small areas of hypopigmentation, which, unless one knew the previous findings, would not have been diagnostic. PMID- 11057399 TI - [Evaluating health education programs for preventing malignant melanoma]. PMID- 11057400 TI - [Good clinical practice. Significance for dermatological research]. PMID- 11057401 TI - Managing medical and insurance information through a smart-card-based information system. AB - The continuously increased mobility of patients and doctors, in conjunction with the existence of medical groups consisting of private doctors, general practitioners, hospitals, medical centers, and insurance companies, pose significant difficulties on the management of patients' medical data. Inevitably this affects the quality of the health care services provided. The evolving smart card technology can be utilized for the implementation of a secure portable electronic medical record, carried by the patient herself/himself. In addition to the medical data, insurance information can be stored in the smart card thus facilitating the creation of an "intelligent system" supporting the efficient management of patient's data. In this paper we present the main architectural and functional characteristics of such a system. We also highlight how the security features offered by smart cards can be exploited in order to ensure confidentiality and integrity of the medical data stored in the patient cards. PMID- 11057402 TI - Using a multihospital systems framework to evaluate and establish drug use policy. AB - PURPOSE: In order to develop rational drug purchasing and use policy for a class of pharmaceuticals used in a consortium system of 14 university based hospitals, the antiemetic use patterns of inpatients receiving cancer chemotherapy were evaluated to assess the comparative effectiveness of granisetron, ondansetron, and conventional antiemetics. PATIENTS AND METHODS: A prospective, observational study was conducted in 14 academic health centers linked under research and purchasing consortium arrangements from October to December 1994. The use of antiemetics was evaluated in hospitalized patients receiving cancer chemotherapy agents with a known propensity for causing, alone or in combination, varying degrees of nausea or vomiting. Clinical outcomes measured were the impact of chemotherapy administration on the functional status of patients, and the occurrence of post-treatment vomiting. RESULTS: The most often prescribed cancer chemotherapy regimens consisted of cisplatin, paclitaxel, etoposide and cyclophosphamide, and the most often prescribed antiemetics were the 5 hydroxytryptamine subtype-3 antagonists (5-HT3 antagonists, granisetron and ondansetron), dexamethasone and lorazepam. Of the 439 patients studied, 329 (75%) reported no episodes of emesis. Of the patients receiving highly emetogenic chemotherapy, those receiving 5-HT3 antagonists experienced better overall outcomes (as measured by functional health status and the absence of vomiting) than patients receiving conventional (non-5-HT3 antagonist) antiemetics. In contrast, patients receiving chemotherapy associated with moderate or low emetogenicity experienced similar outcomes, regardless of the antiemetic regimen selected. No statistical difference was seen between granisetron and ondansetron in achieving positive patient outcomes. CONCLUSION: The study results suggest that 5-HT3 antagonists are associated with better clinical outcomes than other antiemetics in patients receiving highly emetogenic chemotherapy. Less costly conventional antiemetic therapy (or, in some cases, no antiemetic therapy) provide comparable outcomes in patients receiving chemotherapy associated with moderate or low emetogenic potential. Granisetron and ondansetron were found to be clinically comparable. PMID- 11057403 TI - Application of periodogram and AR spectral analysis to EEG signals. AB - In this study, in order to analyze the EEG signal, the conventional and modern spectral methods were investigated. Interpretation and performance of these methods were detected for clinical applications. For this purpose EEG data obtained from different persons were processed by PC computer using periodogram and AR model algorithms. Periodogram and AR modeling approaches were compared for their resolution and interpretation performance. It was determined that the AR approach is better for the use in clinical and research areas, because of the clear spectra that are obtained by it. PMID- 11057404 TI - A method of local skin perfusion detection. AB - On the epidermal surface there are inhomogeneities which manifest as, among other things, lower local impedance. These regions include Head's zones, sweat glands, and also the so-called acupuncture points. The aim of this paper is to examine the effect of skin perfusion on epidermal impedance at the acupuncture points. When measuring epidermis impedance in the local (acupuncture) points the method of impedance plethysmography was applied. Both the method of four electrode measurement at the constant current and the method of two electrode measurement by means of the different and indifferent electrode as well were engaged. First, we had to uncover the size of the action skin points which was made by means of a different micro-electrode developed by us. The effect of some factors on the reproducibility and comparability of the values measured was evaluated. The attention was focused mainly on the questions of the pressure of the different electrode on the skin surface, magnitude and waveform of the exciting current, and so on. The authors demonstrate the impossibility to measure changes in skin impedance in the acupuncture points, in dependence on the blood perfusion--which was due to the high serial resistance between the point measured in the acupuncture place when the four-electrode measurement was applied, on the other side. PMID- 11057405 TI - [Comparison of rhenium-188, rhenium-186-HEDP and strontium-89 in palliation of painful bone metastases]. AB - AIM: Several radiopharmaceuticals were compared previously with regard to the efficiency in pain palliation of bone metastases. Furthermore, first results were reported on the suitability for such kind of therapy of the generator produced radionuclide rhenium-188. METHOD: Influence of Rhenium-188-HEDP (Re-188), Rhenium 186-HEDP (Re-186) and Strontium-89 (Sr-89) on pain symptoms and bone marrow function were obtained in 44 patients (pts). These were 16 pts. with Re-188 (2943 +/- 609 MBq), 13 pts. with Re-186 (1341 +/- 161 MBq) and 15 pts. with Sr-89 (152 +/- 18 MBq) (6 woman with breast cancer and 38 men with prostata cancer). RESULTS: 81 of pts. after Re-188, 77% after Re-186 and 80% after Sr-89 reported relief of pain. The Karnofsky-Index established by pts. increased from 74 +/- 9% to 85 +/- 11% after Re-188, from 70 +/- 11% to 76 +/- 11% after Re-186 and from 62 +/- 10% to 69 +/- 10% after Sr-89. However, the difference between the pre- and the post-therapeutic value is only statistically significant in the case of Re-188 therapy (p = 0.001). A decrease of platelets of 30 +/- 14% after 2.8 +/- 0.7 for pts. treated with Re-188, of 39 +/- 20% after 3.7 +/- 1.0 weeks for pts. treated with Re-186 and of 34 +/- 26% after 4.4 +/- 1.0 weeks for pts. treated with Sr-89 compared to the value before therapy was observed. The difference was not significant between the 3 groups of pts. (p = 0.125 to 0.862). CONCLUSION: All tried radiopharmaceuticals were effective in pain palliation. The various radionuclides had no significant difference in the pain relief or the bone marrow impairment. If only the Karnofsky-Index after Re-188 HEDP seems to be a little more increase. PMID- 11057406 TI - Lymphoscintigraphy in breast cancer patients--comparison of peritumoural and intradermal injection. AB - AIM of this study was to determine whether the sentinel lymph nodes (SLNs) can be accurately identified in breast cancer patients with intradermal injection of the radiotracer above the primary tumour in comparison to peritumoural injection. METHODS: In 45 women with breast cancer we performed lymphoscintigraphy on two separate days. We injected Tc-99m nanocolloid on the first day peritumourally, and on a separate day intradermally. The results of both investigations using different injection sites were compared in order to determine the number and location of SLNs. RESULTS: The SLN identification rate using peritumoural injection was 71% (32 of 45 patients) and 96% (43 out of 45 patients) using intradermal injection. In 62% (28 of 45 patients) the number and location of the SLNs were identical. In 97% (31 of 32 patients) in whom a SLN was detected using peritumoural injection, the same SLNs reappeared with intradermal injection. There were no false negative findings with the peritumoural administration of tracer whereas the intradermal administration approach resulted in a false negative rate of 13%. CONCLUSION: In women with breast cancer the reproducibility of lymphoscintigraphy using peritumoural and intradermal injection sites was 62%. The intradermal injection modality enables the detection of a SLN in patients where the peritumoural injection failed but it has the disadvantage of a higher false negative rate in comparison to the peritumoural injection technique. PMID- 11057407 TI - [Influence of single-photon emission scan duration measured with the ECAT ART PET scanner]. AB - AIM: The aim was to study the influence of single-photon-transmission scan duration in 3D-PET on the quantitative values of attenuation coefficients and noise in transmission images and of activity concentrations and noise in attenuation corrected emission images of thorax phantom- and patient data. METHOD AND MATERIAL: Using dual collimated Cs-137 singles transmission sources (E gamma = 662 KeV, A = 2* 614 MBq) on an ECAT ART tomograph series of transmission scans of a thorax phantom were acquired pre- and post-injection of 18F. 17 patients underwent two transmission scans. The scan time of the short transmission was chosen according to the results of the phantom studies (noise of Poisson statistics less than 4%). Transmission and attenuation corrected emission images were evaluated with respect to estimated linear attenuation coefficients, noise and specific activities in organs. RESULTS: The phantom studies reveal little variation of the estimated linear attenuation coefficients as a function of scan duration. The estimates of the attenuation coefficients are found to be 1% lower than the expected values for pre- and up to 6.5% lower for post-injection transmissions. The noise level in the transmission images increases as expected for Poisson data. The noise level in the attenuation corrected emission images shows only little increase with decreasing transmission scan time whereas it is strongly influenced by a variation of emission scan time. In patient studies, less than 3% difference was found in the estimated linear attenuation coefficients as well as in the activity concentrations between short (pre or post injection) and long transmission scans. The noise levels in transmission and emission images are 1% (pre-injection) and 4% (post-injection) higher for short transmission scans. CONCLUSION: Due to the high photon flux, single photon transmission offers good clinical performance with significantly reduced transmission scan durations (< 2 min/bed in pre-, < 4 min/bed in post-injection transmission). PMID- 11057408 TI - [Clinical value of FDG PET using coincident gamma cameras in staging and restaging of malignant lymphoma--compared with convenitonal diagnostic methods]. AB - AIM of the present retrospective study was to validate the clinical value of F-18 FDG PET imaging in lymphoma patients with a dual head camera modified for coincidence detection. Staging before and after oncological treatment was compared with a conservative diagnostic approach. METHODS: 48 patients (28 non Hodgkin lymphoma, 20 Hodgkin's disease) received FDG-Hybrid-PET scans. Pretherapeutic staging was realized in 28 patients, 9 of them had control studies after they had completed therapy. Totally 29 persons were examined for post therapeutic restaging. Computed tomography imaging and lymph node sonography was performed in all cases. Results were validated by clinical follow-up, in three cases a recidive was proven by biopsy. RESULTS: CT and ultrasound detected 77 lesions in 28 patients compared with 100 visualized by PET, but this difference in pretherapeutic staging did not reach significance at p > 0.05 by Fisher's t test. Hybrid-PET obtained a sensitivity of 93%, a specificity of 79%, a positive of 82% and a negative predictive value of 92% for detection of residual disease. The values for CT + US were 87%, 64%, 72% and 88% respectively. CONCLUSION: FDG Hybrid-PET is as or even more accurate than standard morphologic diagnostic methods for prestaging in malignant lymphoma. Additionally, there is a substantial benefit for therapy monitoring of residual disease using coincidence detection PET with a 3/4-inch crystal gamma camera. PMID- 11057409 TI - [Advantages and limitations of whole-body bone marrow MRI using Turbo-STIR sequences in comparison to planar bone scans] . AB - At modern MRI tomographs the whole body can be screened for bone marrow metastases within 45 min. AIM of this study was to evaluate the diagnostic advantages and disadvantages of a whole-body bone marrow MRI protocol using Turbo Short Tau Inversion Recovery [STIR] sequences in comparison to planar bone scintigraphy (SZ). PATIENTS AND METHODS: In order to screen for bone metastases within two weeks SZ and whole-body MRI with Turbo-STIR-sequences were performed in 20 patients with known breast cancer. For further evaluation five regions were defined: skull, spine including the pelvis, femora, humeri and ribs including scapulae and sternum. RESULTS: In 9/20 patients neither with SZ nor with MRI bone metastases were detected (staging M0). Among the remaining 11 patients SZ detected 109 and MRI 150 lesions which were typical for bone metastases. All of these 11 patients were staged M1 correspondingly with both methods. Within the thorax (ribs, sternum, scapulae) MRI discovered only 6/17 and within the skull 0/6 lesions which were suspicious for metastases in SZ. Inversely MRI identified much more metastatic lesions than SZ within the femora (20/16), the humeri (14/12) and the spine including the pelvis (110/58). CONCLUSIONS: Susceptibility , truncation-, chemical-shift-, third arm- and particularly pulsation artifacts along with the impossibility to chose slice orientation equally advantageous for all regions of the body cause impaired image quality of MRI whole body scanning. Therefore, concerning the detection rate of bone metastases within the thorax (ribs, sternum and scapulae) and the skull, conventional Turbo-STIR-MRI whole body scans are even less accurate than conventional planar bone scintigraphy in those regions. PMID- 11057410 TI - [On the article: Comparison of different methods for attenuation correction in brain PET (Nuclearmedicin 2000; 39: 505)]. PMID- 11057411 TI - Endocrine disruptors: update on xenoestrogens. AB - Endocrine disruptors and their possible impact on human and animal health have become a topic of discussion and an area of active research in toxicology. A focus has been on xenoestrogens, i.e., environmental chemicals with estrogenic activity. In principle, there is agreement that such compounds, in high doses, may cause developmental, reproductive and tumorigenic effects ("hazard"). A matter of controversy is the question of risks associated with xenoestrogens under realistic (low) exposure scenarios; this is due to uncertainty on how to assess the interactions of exogenous compounds with the endocrine system and its complex regulation. Our overview will address topics including: consequences from previous clinical use of the potent estrogen diethylstilbestrol with particular emphasis on dose-response relationships, other observations in humans exposed to estrogenic chemicals in an occupational context, and available information on exposure levels of synthetic and naturally occurring estrogens in the diet. Together with a critical appraisal of methods to detect and quantitate the estrogenic activity of synthetic and naturally occurring chemicals, novel aspects in the risk assessment for endocrine active compounds are discussed. PMID- 11057412 TI - Long-term monitoring of sister chromatid exchanges and micronucleus frequencies in pharmacy personnel occupationally exposed to cytostatic drugs. AB - OBJECTIVES: Many antineoplastic drugs were found to have carcinogenic, mutagenic and teratogenic potential. The aim of this study was to carry out cytogenetic and internal dose monitoring of hospital pharmacy personnel regularly involved in the preparation of cytostatic agents, in order to test possible cytostatics-induced genotoxic effects due to occupational exposure under routine working conditions, and in cases of accidental contamination. METHODS: Platinum in whole blood and anthracyclines in plasma were measured to assess internal exposure to cytostatics. The level of cytogenetic damage was determined in peripheral blood lymphocytes with the micronucleus test and the sister chromatid exchange assay. Five series of monitoring were performed over a period of 2 years. RESULTS: No significant differences in the mean frequencies of sister chromatid exchanges (SCE) and micronuclei (MN) were found between occupationally exposed probands and controls (9.9 +/- 1.4 vs 10.1 +/- 1.2 SCEs/cell and 21.2 +/- 7.2 vs 23.3 +/- 7.5 MN/2000 binucleated (BN) cells, n = 16). Significant elevations of SCE or MN were detected in seven out of 12 cases of accidental contamination at the workplace, whereas no increase in platinum in blood and anthracyclines in plasma was observed in these probands. Two cases of non-reported contamination were identified by measurement of epirubicin in plasma. Smoking was found to increase the SCE significantly. No correlation between individual SCE scores and MN scores was observed. CONCLUSIONS: Our findings support a transient increase in SCE or MN after relevant exposure to cytostatic drugs in cases of accidental contamination. The lack of significant differences in SCE and MN between hospital pharmacy personnel and unexposed controls, points to high standards of safety at the corresponding workplaces. PMID- 11057413 TI - Possible metabolic interaction between hexane and other solvents co-exposed at sub-occupational exposure limit levels. AB - OBJECTIVE: To investigate whether metabolic interactions exist between hexane (HEX) and other solvents when co-exposed at the levels below occupational exposure limits. METHODS: Workers, 219 men in ten workshops in total, volunteered to participate in the study. They were occupationally exposed to mixtures of HEX and one or more of toluene (TOL), ethyl acetate (EA) and acetone (ACE). Time weighted average intensity of vapor exposures was monitored by diffusive personal sampling. 'Free'- and 'total'-2,5-hexanedione (HD) levels in the end-of-shift urine samples were determined by gas chromatography (GC) before and after acid hydrolysis of urine, respectively, and expressed as observed (HDob) or after correction for creatinine concentration (HDcr) or urine specific gravity (HDsg). Possible interaction was examined by multiple regression analysis (MRA), taking either free- or total-HD as a dependent variable, and the four solvent concentrations as independent variables. RESULTS: In most cases, exposure intensity did not exceed the current occupational exposure limits even when additiveness was assumed. In addition that HEX was the most influential independent variable in all cases as expected, the MRA showed that, in cases of free-HD, ACE was also influential to HDob although weakly, but not to HDcr or HDsg. With regard to total-HD, ACE was weakly influential to HDob and HDsg, and EA also weakly to HDcr. The effect of ACE on free- or total-HD was not detected, however, when 22 men exposed only to HEX and ACE were subjected to the same analysis. Similarly, the effect of EA on total-HD was not observed among the remaining 197 men exposed to HEX, TOL and EA only. CONCLUSIONS: When the exposures were below occupational exposure limits, the free-HD levels in urine after HEX exposure will not be modified by co-exposures to TOL, EA or ACE. PMID- 11057414 TI - Saliva biomonitoring of atrazine exposure among herbicide applicators. AB - A field study was conducted in which saliva samples were collected from a cohort of herbicide applicators during the pre-emergent spray season in Ohio in 1996. Atrazine concentrations were detected in human saliva samples using an enzyme linked immunosorbent assay (ELISA) method. Trend due to atrazine exposure and subsequent elimination in the body were evidenced by the temporal pattern of decreasing atrazine concentrations in saliva over time. Median salivary concentrations of atrazine on non-spray days were significantly lower than on spray days for each sampling time (Mann-Whitney U-Wilcoxon rank sum test, P < 0.01). Within spray days, median salivary atrazine concentrations were significantly higher on days atrazine was sprayed than on days herbicides other than atrazine were sprayed for each sampling time (Mann-Whitney U-Wilcoxon rank sum test, P = 0.02 for 4 6 p.m. samples, P = 0.04 for bedtime samples, P = 0.03 for next-morning samples). Median salivary atrazine concentrations on days atrazine was sprayed were higher than the median concentration for the corresponding sampling time on non-spray days and on days when other herbicides were sprayed. Salivary concentration of atrazine is a plausible indicator of those days in which atrazine spraying was likely to have occurred. Salivary concentrations of atrazine not only reflect exposures resulting from spraying atrazine, but also exposures from other field activities where applicators may come in contact with atrazine. The results of this study confirmed data from animal experiments that atrazine is able to cross the cell membranes of salivary glands, and can be measured in human saliva with high sensitivity. The sampling method itself is convenient and easy to use in the field, with a high compliance rate, and analytical procedures are rapid and inexpensive. It is, therefore, concluded that saliva sampling of atrazine exposure among herbicide applicators is a feasible biomonitoring method. PMID- 11057415 TI - Biological monitoring of occupational exposure to N,N-dimethylformamide--the effects of co-exposure to toluene or dermal exposure. AB - OBJECTIVES: The objective of this study is to assess the exposure and intake dose of N,N-dimethylformamide (DMF) and the correlation between them, according to the type of exposure for the workers in the DMF industry. METHODS: We monitored 345 workers occupationally exposed to DMF, from 15 workshops in the synthetic fiber, fiber coating, synthetic leather and paint manufacturing industries. Ambient monitoring was carried out with personal samplers to monitor the external exposure. Biological monitoring was done to determine the internal dose by analyzing N-methylformamide (NMF) in end-shift urine. Work procedure and exposure type of each DMF workshop was carefully surveyed, to classify workers by exposure type according to work details. Workers were classified into three groups (Group A: continuous and direct exposure through inhalation and skin; Group B: intermittent and short-term exposure through inhalation and skin; Group C: continuous and indirect exposure mostly through inhalation). RESULTS: Geometric mean of DMF concentration in air was 2.62 (GSD 5.30) ppm and that of NMF in urine was 14.50 (GSD 3.89) mg/l. In the case of continuous absorption through inhalation and dermal exposure (Group A), the value of NMF in urine corresponding to 10 ppm of DMF was 45.3 mg/l (r = 0.524, n = 178), 39.1 mg/g creatinine (r = 0.424), while it was 37.7 mg/l (r = 0.788, n = 37), 24.2 mg/g creatinine (r = 0.743) in the case of absorption mostly through inhalation (Group C). Creatinine correction reduced the correlation between two parameters. CONCLUSION: The NMF in urine corresponding to 10 ppm DMF, of the dermal and inhalation exposure group was 39.1 mg/g creatinine (r = 0.424, n = 178), while that of the inhalation exposure-only group was 24.2 mg/g creatinine (r = 0.743, n = 37). Co-exposure with toluene reduced the NMF excretion in urine. PMID- 11057416 TI - Applicability of homogeneous exposure groups for exposure assessment in the chemical industry. AB - The purpose of the study was to assess occupational exposure to chemicals, by taking the aspects presented in European standard EN 689 into account, especially with respect to homogeneous exposure groups and their suitability as the basis for exposure assessment. In addition, dermal exposure to chemicals was assessed when appropriate. The industries studied included a plywood factory, a paint factory and a sewage treatment plant of an oil refinery. The workers were classified into homogeneous exposure groups by the persons in charge of occupational health issues in the respective workplaces. The concentrations of the contaminants were measured in workplace air by breathing-zone and stationary sampling, and these approaches were compared. Dermal exposure was measured when applicable. The homogeneity of the grouping was tested with analysis of variance whenever possible. The tasks studied in plywood manufacturing fulfilled the criteria of homogeneous exposure groups for both respiratory and dermal exposure. The group of operators in the sewage treatment plant was highly homogeneous. The complicated organization of the tasks made the use of homogeneous exposure groups (HEG) unsuitable at the paint factory. These findings show that reliable exposure assessment cannot be achieved with a formal standard; instead, comprehensive occupational hygiene evaluation is needed. It should also have a great importance when exposure models are developed. PMID- 11057417 TI - Toxicokinetic modelling of methyl formate exposure and implications for biological monitoring. AB - A toxicokinetic (TK) model was developed to describe the inhalation exposure in humans to methyl formate (MF), a catalyst used in foundries, and to discuss biological monitoring. The TK model consisted of four compartments: MF, the metabolites--methanol (MeOH) and formic acid (FA)--and, in addition, a urinary compartment describing the saturable reabsorption of FA. Levels of MeOH and FA in urine, from an experimental study (100 ppm MF, 8 h at rest), validated the present model. The TK model describes well the general behaviour of MeOH and FA in urine after MF exposure. A nonlinear and a linear relationship respectively, was predicted between MF exposure and FA or MeOH excretion in urine, and this has previously been seen after occupational MF exposure. The present model has been modified to simulate MeOH exposure as well. Generally low exposures (concentration or exercise) produce only marginal increases in FA urinary excretions, but when exposure is elevated, urinary FA excretion increases because of saturation in the mechanism of reabsorption. Using FA urinary excretion as the critical indicator, because of its link to health effects, an occupational exposure limit value for MF of no greater than 50 ppm should be selected (based on predictions with the TK model). MeOH in urine can be considered as a biomarker for MF at low exposure, because of lower background values and of a linear relationship with exposure. At higher exposures, however, FA could be used as a biomarker as it becomes progressively more sensitive. But the use of biological monitoring for MF is difficult because of individual variations in background values. Under the present state of knowledge both FA and MeOH should be used to estimate only group exposures, rather than individual exposures. PMID- 11057418 TI - Nasal and bronchial responses to flour-inhalation in subjects with occupationally induced allergy affecting the airway. AB - OBJECTIVES: The aim of the study was to follow the similarities and differences, of cellular and mediator changes and mucosal/vascular permeability in the upper and lower airway after specific and nonspecific bronchial provocation, in bakers with diagnosed occupationally induced allergy affecting the airway. In addition, the authors try to find whether there is a relationship between cellular changes in nasal and bronchoalveolar lavage, and bronchial hyperreactivity. METHODS: The study participants were 10 bakers with occupational bronchial asthma and allergic rhinitis. All patients were sensitized to investigated allergen-flour. Nasal- and bronchoalveolar lavage techniques were used to evaluate the changes of the cellular and mediator response (tryptase, eosinophil cationic protein, ECP) and albumin level after specific (flour) and placebo provocation. In addition. bronchial hyperreactivity for histamine, and forced expiratory volume in 1 s (FEV1) were measured after the challenge. RESULTS: There was a significant increase in the percentage of eosinophils, basophils and albumin in nasal and bronchoalveolar lavage of occupationally sensitized bakers. A statistically significant increase in the percentage of neutrophils in bronchoalveolar lavage was observed only 24 h after the allergen challenge. The level of tryptase in nasal lavage was significantly higher during the early allergic response. The levels of ECP in both nasal and bronchoalveolar lavage were significantly increased during the late allergic response. There were also severe bronchial reactions and increase of bronchial hyperreactivity for histamine in occupationally sensitized bakers in the late phase of allergic reaction. CONCLUSION: Eosinophils and basophils proved to be the predominant cells in nasal and bronchoalveolar lavage of patients with occupationally induced bronchial asthma and rhinitis. The prolonged increase of albumin level seems also to be a good predictor of protracted nasal and bronchial inflammation. The results obtained confirmed that tryptase and ECP are good markers for monitoring mast cell and eosinophil degranulation during the allergic reaction. Increase of airway responsiveness reflects an eosinophil and basophil contribution to airway allergic response. PMID- 11057419 TI - Powdering floor polish and mucous membrane irritation in secondary school pupils. AB - OBJECTIVES: Acrylate-styrene copolymer polish has been used to protect the surface of linoleum flooring since the 1960s. Problems with powdering of floor polish were observed at an early stage. In a secondary school in Linkoping, Sweden, this phenomenon occurred in the winter of 1994-1995 and the pupils frequently reported irritative symptoms from the eyes and airways. This study was undertaken to assess the potential effect of powdering floor polish on the mucous membranes of the eyes and respiratory tract. METHODS: Repeated questionnaire based surveys were conducted with identical questions in the spring of 1995 (during the powdering period) and in the autumn of 1995 (after the polish was removed). The questions dealt with irritative symptoms from the nose, eye, throat and lower respiratory tract. RESULTS: A preventive effect related to the removal of polish was found for irritative symptoms in all locations mentioned above, but was particularly clear for the lower respiratory tract (prevalence rate ratio = 0.37, 95% CI = 0.23-0.59). CONCLUSIONS: The results of this study indicate that the powdering of floor polish may cause irritative symptoms from the eyes and airways in school children. PMID- 11057420 TI - Antioxidative stress response in workers exposed to carbon disulfide. AB - OBJECTIVE: To study the effects of carbon disulfide (CS2) on antioxidative stress systems of exposed workers. METHODS: Enzyme-linked immunosorbent assay (ELISA) and thiobarbituric acid test (BAT) were conducted on 67 exposed workers and 88 controlled ones in a viscose rayon factory to determine their serum cuprozinc superoxide dismutase (CuZnSOD) and malonyldialdehyde (MDA) levels. RESULTS: The average levels of CuZnSOD in workers exposed to CS2 both above and below 10 mg/m3 were higher than those in the control group (P < 0.0001), showing some dose effect and dose-response relationships. SOD levels increased when the exposure index (EI) was less than 300, and remained at a high level at the range of 300 to 900. When EI was higher than 900, SOD tended to decrease. Meanwhile, the serum MDA levels increased. Both CS2 concentrations and exposure time contribute to the MDA levels. CONCLUSIONS: CS2 exposure could influence the stress response of the oxidative-antioxidative system of workers. Increased SOD levels could be considered as the stress response of antioxidative system to CS2 exposure in the early stages, and the influence of CS2 on SOD might be bi-directional. SOD and MDA might become objective indices in workers' health surveillance. The role of these two indices in the intoxication mechanism still needs to be clarified. PMID- 11057421 TI - Ambulatory blood pressure monitoring in children and adolescents: the future. PMID- 11057422 TI - Silent ischaemia and hypertension. AB - For many years now, silent ischaemia has been recognized as a distinct clinical entity, and its relevance in different patient groups has been established. However, a number of basic questions have not been answered. In explaining the pathophysiology of silent ischaemia, factors affecting both the demand and the supply side are now being recognized. With the exception of certain well-defined groups, it is not clear why some patients are mostly symptomatic, while other patients are predominantly asymptomatic. There appear to be many factors influencing the ischaemic pain threshold. Studies investigating the prevalence of silent ischaemia show a remarkably high prevalence of silent ischaemia in different patient groups. Patients with hypertension but without coronary artery disease form a specific and vulnerable high-risk population that is particularly prone to silent ischaemia. Since changes at the macrovascular level are not responsible, various factors negatively influencing either cardiac supply or demand have been investigated. A reduced coronary reserve is central in explaining the increased prevalence of silent ischaemia in hypertensives. Left ventricular hypertrophy renders meaningful detection of ST segment changes difficult, but a possible solution dealing with this problem is offered by applying more stringent criteria in terms of minimal ST depression in the definition of ischaemia. The treatment of silent ischaemia is largely the same as for angina pectoris, but whether therapy should be directed at elimination of all ischaemic episodes or only of symptomatic episodes depends on further prospective work addressing this question. PMID- 11057423 TI - Chronic and acute effects of oestrogens on vascular contractility. AB - In addition to their role as sex hormones, it has been known for many years that oestrogens have protective effects on the vasculature. These have been implicated in the reduced incidence of cardiovascular disorders in premenopausal women and in post-menopausal women receiving oestrogen replacement therapy. This protection has been found to be due, in part at least, to direct effects of oestrogens on blood vessels. This review will summarize the available literature regarding oestrogenic effects on vascular contractility. Two major influences of oestrogens will be discussed; first the genomic effects induced by chronic administration of steroid hormones, and second, the rapid effects on vascular smooth muscle by non genomic, and as yet not fully identified, mechanisms. In so doing, the diversity of oestrogenic actions on vascular contractility will be highlighted and the protective role of these agents against adverse cardiovascular events discussed. PMID- 11057424 TI - Multiple risk factor clustering of hypertension in a screened cohort. AB - OBJECTIVE: A family history of hypertension, obesity, diabetes mellitus, hypercholesterolaemia and hypertriglyceridaemia have all been associated with the risk for hypertension. We evaluated whether the clustering of these risk factors increases the risk for hypertension or whether the accumulation of risk factors is associated with the blood pressure level in non-hypertensive subjects. METHODS AND SUBJECTS: We assessed the clinical data and family history of hypertension (in parents and siblings) for 9914 individuals (6163 men and 3751 women, 18-89 years old) who were screened in Okinawa, Japan, in 1997. RESULTS: In 9914 subjects (2465 hypertensive and 7449 non-hypertensive subjects), all the five factors were positively associated with hypertension. The odds ratios (95% confidence interval) for the number of risk factors were 1.88 (1.62-2.18) for one risk factor, 3.06 (2.62-3.57) for two, 5.25 (4.37-6.30) for three, 8.71 (6.48 11.72) for four and 24.48 (8.49-70.56) for five, after adjusting for age, sex, alcohol consumption, cigarette smoking and physical exercise habits. In non hypertensive subjects, multivariate regression analyses showed that the number of risks was positively correlated with blood pressure; the regression coefficient was 1.96 (P < 0.0001) for systolic blood pressure, and 1.47 (P < 0.0001) for diastolic blood pressure after adjusting for age and sex. CONCLUSIONS: Clustering of risk factors was significantly associated with hypertension. The number of risk factors positively correlated with the blood pressure levels in nonhypertensive subjects. The accumulation of risk factors may play an important role in the pathogenesis of hypertension, and thus the aggregation of risk factors may need to be addressed in primary prevention efforts related to hypertension. PMID- 11057425 TI - Temporal changes in clinic and ambulatory blood pressure during cyclic post menopausal hormone replacement therapy. AB - OBJECTIVE: Post-menopausal hormone replacement (HRT) might protect against cardiovascular disease, possibly by arterial vasodilation and reduced blood pressure. Progestogens are needed to avoid endometrial disease but vascular effects are controversial. The objective was to assess temporal changes in blood pressure (BP) by two measurement techniques during a cyclic hormone replacement regimen. DESIGN AND METHODS: Sixteen healthy and normotensive post-menopausal women (age 55 +/- 3 years) were studied in a placebo-controlled, randomized crossover study, and were randomized to 17beta-oestradiol plus cyclic norethisterone acetate (NETA) or placebo in two 12-week periods separated by a 3 month washout Clinic blood pressure was measured sitting by the same observer with a mercury manometer at four visits in each period. Twenty-four hour ambulatory blood pressure was measured at baseline and in the ninth weeks of treatment in both periods. RESULTS: Clinic systolic and diastolic BP were reduced after 10 days of oestradiol (-5.1 and -3.2 mmHg respectively, P < or = 0.05). After 9 weeks of cyclic HRT, prior to progestogen addition, clinic BP returned to baseline. During addition of NETA, diastolic blood pressure was again reduced ( 3.6 mmHg, P= 0.037). Mean 24 h ambulatory systolic and diastolic blood pressures were significantly lower than clinic measurements (-15.7 and -5.9 mmHg, P < 0.001) but were unaffected by HRT. CONCLUSIONS: Clinic blood pressure is reduced during a cyclic HRT regimen but the reduction varies with the HRT regimen, which might explain the diversity in previous BP findings during HRT. Norethisterone acetate might possess additive blood pressure-lowering effects in postmenopausal women. PMID- 11057426 TI - The lack of a modulating effect of non-genetic factors (age, gonads and maternal environment) on the phenotypic expression of the salt-susceptibility genes in the Sabra rat model of hypertension. AB - OBJECTIVE: This study was designed to test the hypothesis that non-genetic factors such as age, gonads and maternal environment modulate the expression of the salt-susceptibility genes and affect the blood pressure response to salt loading (salt-sensitivity and salt-resistance) in the Sabra rat model of hypertension. METHODS: The blood pressure response to salt-loading was studied in Sabra hypertension prone (SBH/y) and Sabra hypertension resistant (SBN/y) rats of both sexes: (1) at 1, 3, 6, 9 and 12 months of age, (2) in adult rats after orchiectomy or oophorectomy, and (3) in animals that had been raised and nourished from birth to weaning by a foster mother from the contrasting strain. In each of the study protocols, systolic blood pressure was measured at baseline by the tail cuff method, animals were salt-loaded with deoxycorticosterone acetate, and blood pressure was measured again after 4 weeks. RESULTS: Basal blood pressure at all the ages studied and in both sexes was on average 10-15 mmHg higher in SBH/y than in SBN/y. Salt-loading in SBN/y of both sexes aged 1-12 months did not induce any significant increment in blood pressure. Salt-loading in SBH/y, in contrast, caused a highly significant rise in systolic blood pressure, of 40 mmHg or more at all the ages studied. There was no age difference or sex dependence in the magnitude of the blood pressure response to salt Oophorectomy or orchiectomy did not affect the levels of basal blood pressure nor prevent the hypertensive response to salt-loading in SBH/y or the lack of a hypertensive response in SBN/y rats. Gonadectomy did not affect blood pressure in salt-loaded hypertensive SBH/y nor in salt-loaded normotensive SBN/y. The basal blood pressure and the blood pressure responses of SBH/y and SBN/y of both sexes raised by foster mothers of the contrasting strains from birth to weaning were not different from those observed when raised by their natural mothers. CONCLUSIONS: This study indicates that salt-sensitivity in SBH/y and salt resistance in SBN/y are not age-dependent phenomena; that the magnitude of the BP response to salt-loading is not sex-dependent; and that neither gonadectomy nor the maternal environment affect the blood pressure response to salt-loading in the adult animal of either strain. These non-genetic factors thus do not modulate expression of the salt-susceptibility genes in the Sabra genetic model of salt sensitive hypertension. PMID- 11057427 TI - The renin-angiotensin-aldosterone system and carotid artery disease in mild-to moderate primary hypertension. AB - BACKGROUND: The evidence linking activation of the renin-angiotensin system with accelerated cerebro-vascular atherosclerosis remains controversial. We therefore prospectively investigated the relationships of plasma renin activity and aldosterone levels with carotid artery lesions (CAL) in essential hypertension. METHODS: We evaluated the prevalence and severity of CAL and the intimal-medial thickness (IMT) with a high-resolution echo-Doppler technique in 107 cerebrovascularly asymptomatic consecutive primary hypertensives (55 male, 52 female) and in 70 (42 male, 28 female) normotensive controls. We also measured supine plasma renin activity (PRA) and aldosterone before and 45 min after captopril administration, while daily urinary excretion of sodium was measured. RESULTS: Both the prevalence (59.4 versus 26.2%) and severity of sex- and age adjusted and unadjusted CAL and IMT were significantly higher in hypertensives than in controls. Regression analysis showed different predictors of IMT (age and captopril-stimulated-PRA, R2 = 0.27, P < 0.0001), score of CAL (mean blood pressure, R2 = 0.15, F=12.73, P< 0.0001) and maximal stenosis (pulse pressure and known duration of hypertension R2 = 0.29, F = 14.58, P< 0.0001). Sex- and age adjusted IMT did not differ between quartiles of renin-sodium profile. However, patients in the quartile with the highest PRA had the lowest score of CAL and an inverse relationship between age-adjusted PRA and IMT and CAL was found. CONCLUSIONS: These results, besides confirming an association of both IMT and CAL with primary hypertension and ageing, demonstrate that CAL and IMT have different correlates. However, they do not support the contention that a high renin-sodium profile carries an excess risk of CAL in primary hypertensives with no clinical evidence of cerebro-vascular disease. PMID- 11057428 TI - Anti-apoptotic action of hepatocyte growth factor through mitogen-activated protein kinase on human aortic endothelial cells. AB - OBJECTIVE: To investigate the molecular mechanisms of the anti-apoptotic action of hepatocyte growth factor (HGF), a novel angiogenic growth factor that may have a pivotal role in the regulation of endothelial cells, on human aortic endothelial cells. METHODS: An index of cell number and death was determined using a water-soluble tetrazolium salt dye assay, DNA fragmentation enzyme-linked immunosorbent assay, and non-confocal fluorescence microscopy of nuclear staining with Hoechst 33258 and propidium iodide. Extracellular-signal-regulated protein kinase (ERK) and the p38 mitogen-activated protein kinase (p38MAPK) were analysed by Western blotting using a phospho-specific antibody. RESULTS: Treatment of quiescent endothelial cells with HGF resulted in significant dose-dependent increases in cell numbers and decreases in lactate dehydrogenase (LDH) release. Moreover, HGF significantly attenuated endothelial cell death induced by culture in serum-free conditions. We therefore focused on the signal transduction system, and in particular on ERK and p38MAPK. ERK was markedly phosphorylated by HGF. The contribution of ERK to cell growth was supported by the observation that addition of PD98059, a specific inhibitor of MAPK kinase, significantly attenuated the increase in endothelial cell numbers induced by HGF, in a dose-dependent manner. Similarly, PD98059 also attenuated the decrease in LDH release and DNA fragmentation by HGF under serum-free conditions. Interestingly, ERK was re phosphorylated at 12 h after stimulation. Re-phosphorylation of ERK was the result of induction of endogenous HGF by exogenously added HGF, as addition of neutralizing anti-HGF antibody to the conditioned medium attenuated re phosphorylation of ERK at 12 h. In contrast, although p38MAPK was also phosphorylated by HGF, SB203580, a specific inhibitor of p38MAPK, failed to change the endothelial cell growth induced by HGF. CONCLUSION: We have demonstrated that the anti-apoptotic action of HGF against endothelial cell death was mainly through phosphorylation of ERK on human endothelial cells. PMID- 11057429 TI - Birth weight relates to blood pressure and microvascular function in normal subjects. AB - OBJECTIVE: The relationship between low birth weight and elevated blood pressure in adult life is well established but presently unexplained. Both microvascular dysfunction and insulin resistance have been proposed as a possible explanation. We have examined the relation between birth weight and blood pressure in 30 healthy subjects exhibiting a wide range of insulin sensitivity, and assessed whether microvascular function and/or insulin resistance may underlie this relationship. METHODS: Birth weight data were obtained from birth announcements. Blood pressure was measured with an ambulatory blood pressure monitor and insulin sensitivity was assessed by the hyperinsulinaemic, euglycaemic clamp technique. Microvascular function, i.e. capillary recruitment and endothelium-dependent and independent vasodilatation in the skin, was evaluated by videomicroscopy and iontophoresis of acetylcholine and sodium nitroprusside. RESULTS: Birth weight was significantly associated with blood pressure (r= -0.50; P< 0.05), capillary recruitment (r= +0.52; P< 0.05), acetylcholine-mediated vasodilatation (r= +0.40; P< 0.05), insulin sensitivity (r= +0.62; P< 0.01) and waist-to-hip ratio (r= 0.42; P< 0.05). Regression analysis showed a significant association of birth weight with 24 h systolic blood pressure (regression coefficient: -7.6 mmHg/kg; 95% confidence interval: -13.0 to -1.0). Adjustment for capillary recruitment and waist-to-hip ratio decreased the regression coefficient by 39 and 41%, respectively. The results were similar after adjustment for age, sex or body mass index. CONCLUSION: These results suggest that capillary recruitment and body fat distribution may partly explain the relationship between birth weight and blood pressure. PMID- 11057430 TI - The calcium-channel blocker lacidipine reduces the development of atherosclerotic lesions in the apoE-deficient mouse. AB - BACKGROUND: Lacidipine is a widely used calcium-channel blocker, which has both long-lasting antihypertensive activity and also antioxidant properties. Previous studies have demonstrated the ability of lacidipine to reduce the development of atherosclerotic lesions in several animal models. OBJECTIVE: The present study investigated the antiatherosclerotic potential of lacidipine in the apoE deficient mouse, an experimental model of atherosclerosis showing progressively complex and widespread lesions which closely resemble the inflammatory-fibrous plaques seen in humans. METHODS: Lacidipine was administered daily by gavage for 10 weeks at dose levels of 0 (control), 0.3, 1.0 and 3.0 mg/kg. RESULTS: Lacidipine administration reduces the extension of atherosclerotic lesions in the aorta of the apoE-deficient mouse without affecting plasma lipid levels. We also show that apoE-deficient mice have four-fold higher values of the proatherogenic peptide, endothelin, compared with the wild-type C57BL/6 mouse and that lacidipine administration reduced, in a dose-dependent manner, the concentrations of plasma endothelin. CONCLUSION: Lacidipine has anti-atherogenic effects in the apoE-deficient mouse, and reduces plasma endothelin concentrations. PMID- 11057431 TI - Hypertension associated with multiple renal arteries may be renin-dependent. AB - OBJECTIVE: Subjects with multiple renal arteries have been shown to suffer more frequently from hypertension and to have higher blood pressures than subjects whose kidneys are supplied by single renal arteries. This study was carried out to determine whether subjects with multiple renal arteries also have higher renin activity. METHODS: We studied 62 consecutive patients who had undergone angiography for various reasons. They were divided into two groups. Group A comprised 29 patients whose kidneys were supplied by single arteries (male :female ratio 1.63, mean age 51.8 +/- 1.9 years) while Group B comprised 33 patients with multiple renal arteries (male:female ratio 2, mean age 47.3 +/- 2.3 years). RESULTS: Before stimulation with frusemide, the plasma renin in Group A was 0.79 +/- 0.13 ng angiotensin l/ml per h, while in Group B the corresponding figure was 1.73 +/- 0.38 ng angiotensin l/ml per h. This difference was statistically significant (P= 0.0127). Thirty minutes later the plasma renin level in Group A was 2.43 +/- 0.37 ng angiotensin l/ml per h versus a level of 3.86 +/- 0.53 ng angiotensin l/ml per h in Group B (P= 0.0169). Again, 30 minutes later the level was 2.59 +/- 0.4 ng angiotensin l/ ml per h in Group A, versus 3.79 +/- 0.59 ng angiotensin l/ ml per h in Group B (P= 0.0495). CONCLUSIONS: We conclude that patients with multiple renal arteries constitute a group who have high plasma renin activity and may therefore be prone to develop arterial hypertension. PMID- 11057432 TI - Metformin treatment corrects vascular insulin resistance in hypertension. AB - OBJECTIVE: In states of insulin resistance, the vasorelaxant actions of insulin are blunted, which may contribute towards the development of increased vascular tone/hypertension and reduced glucose uptake. To examine whether treating insulin resistance in hypertension restores the vascular actions of insulin, we studied the long-term effects of metformin on the contractile responses of isolated aortas from control and insulin-resistant, hyperinsulinaemic fructose hypertensive rats in the presence and absence of insulin. DESIGN AND METHODS: Sprague Dawley rats were divided into control, control metformin-treated, fructose and fructose metformin-treated groups (n = 8 per group). The treated groups received metformin (500 mg/kg per day for 6 weeks), following which isometric responses to noradrenaline (NA) and angiotensin II (A-II) were examined in thoracic aortas in the presence and absence of insulin (100 mU/ml for 2 h) using isolated organ-bath apparatus. In addition, endothelium-dependent and independent vascular responses to acetylcholine (ACh) and sodium nitroprusside (SNP) were also studied. RESULTS: Metformin treatment prevented the development of fructose-induced insulin resistance, hyperinsulinaemia and hypertension. Insulin attenuated the contractile responses to NA and A-II in control rat aortas; however, blood vessels from untreated fructose rats were refractory to insulin-induced vasodilation. Strikingly, long-term metformin treatment restored the vasodepressor actions of insulin in fructose rats. Metformin did not affect the contractile responses to NA or A-II in either control or fructose rats. In addition, metformin treatment restored ACh-induced endothelium-dependent vasorelaxation in aortas from fructose rats without affecting SNP-induced relaxation. CONCLUSIONS: These data show, for the first time, that long-term metformin treatment corrects vascular insulin resistance and improves endothelium dependent vasorelaxation in hypertension. These effects appear to be secondary to metformin-induced improvements in metabolic derangements (versus a direct vascular action of metformin). Improving the vascular effects of insulin may serve to decrease peripheral tone, attenuate blood pressure and improve insulin sensitivity. PMID- 11057433 TI - Effects of a low-energy diet and an insulin-sensitizing agent on ambulatory blood pressure in overweight hypertensive patients. AB - OBJECTIVE: To clarify the role of insulin resistance and hyperinsulinaemia in the pathogenesis of obesity-related hypertension. DESIGN: An open study comparing the effects of weight reduction by low-energy diet and treatment with troglitazone, an insulin-sensitizing agent. SETTING: A tertiary teaching hospital. PATIENTS: Thirty overweight hypertensive patients (15 men and 15 women, mean age 61 years, mean body mass index 29.1 kg/m2). INTERVENTIONS: Fifteen patients were assigned to a weight-reduction programme by low-energy diet (3360 kJ/day) for 3 weeks; the remaining 15 patients were treated with troglitazone (400 mg/day) for 8 weeks. MAIN OUTCOME MEASURES: Casual and ambulatory blood pressures, glucose and lipid metabolism, and insulin sensitivity. RESULTS: The baseline values of body mass index, fasting and post-glucose plasma insulin, and casual and ambulatory blood pressures were comparable between the two groups. Weight reduction (4.1 +/- 0.3 kg, mean +/- SEM) was associated with significant decreases in plasma insulin, blood glucose, homeostasis model assessment (HOMA) insulin resistance index, serum triglyceride, casual blood pressure (7.7 +/- 2.3/ 3.9 +/- 1.4 mmHg) and 24 h blood pressure (8.3 +/- 1.9/ 4.3 +/- 1.1 mmHg). Treatment with troglitazone caused comparable decreases in the metabolic parameters and HOMA index, but did not change casual or 24 h blood pressure (0.8 +/- 3.4/0.8 +/- 2.1 and 1.5 +/- 2.4/ 1.0 +/- 1.9 mmHg, respectively). CONCLUSIONS: Insulin resistance/hyperinsulinaemia may not have an important role in the pathogenesis of obesity-related hypertension. The antihypertensive effect of weight reduction seems to be mediated mainly by other mechanisms. PMID- 11057434 TI - Glucose modifies the cross-talk between insulin and the beta-adrenergic signalling system in vascular smooth muscle cells. AB - BACKGROUND: Abnormalities in the vascular function of insulin are observed in insulin resistance, and hyperglycaemia is one of the important factors inducing insulin resistance. OBJECTIVE: To investigate the role of glucose in the interaction of insulin and beta-adrenergic signalling systems in vascular smooth muscle cells (VSMC). METHODS: After cells were treated with D-glucose (525 mmol/l) and insulin (100 nmol/l), adenylyl cyclase activity was measured in the presence of isoproterenol, forskolin, and cholera toxin. Assays for insulin induced activities of insulin receptor substrate (IRS)-1, phosphoinositide 3 kinase (PI3-K) and mitogen-activated protein kinase (MAPK) were performed. RESULTS: In the presence of low glucose concentrations (5 mmol/l), insulin enhanced isoproterenol-, forskolin- and cholera toxin-stimulated adenylyl cyclase activities. This stimulatory effect was abolished by PI3-K inhibitors, wortmannin, or LY294002. In contrast, in the presence of high glucose concentrations (25 mmol/l), insulin attenuated isoproterenol-stimulated activity but not cholera toxin- or forskolin-stimulated activity. Insulin-stimulated activities of IRS-1 and PI3-K, but not MAPK activity, were also attenuated in the presence of high concentrations of glucose. The MAPK kinase inhibitor, PD98059, abolished the inhibitory effect of insulin on the beta-adrenergic signalling system. Troglitazone and pioglitazone prevented this inhibitory effect of insulin by restoring IRS-1 and PI3-K activities. CONCLUSIONS: In the presence of low glucose concentrations, insulin stimulates the beta-adrenergic signalling system through the IRS-1/PI3-K pathway. However, in the presence of high glucose concentrations, the effect of insulin is switched to an inhibitory one, through the MAPK pathway. Our finding suggests that high glucose concentrations modify the cross-talk between insulin and the beta-adrenergic signalling systems in VSMC. PMID- 11057435 TI - Regression of left ventricular hypertrophy in hypertensive patients treated with indapamide SR 1.5 mg versus enalapril 20 mg: the LIVE study. AB - OBJECTIVE: To compare the efficacy of indapamide sustained release (SR) 1.5 mg and enalapril 20 mg at reducing left ventricular mass index (LVMI) in hypertensive patients with left ventricular hypertrophy (LVH). DESIGN: The LIVE study (left ventricular hypertrophy regression, indapamide versus enalapril) was a 1 year, prospective, randomized, double-blind study. For the first time, a committee validated LVH before inclusion, provided on-going quality control during the study, and performed an end-study reading of all echocardiograms blinded to sequence. SETTING: European hospitals, general practitioners and cardiologists. PATIENTS: Hypertensive patients aged > or = 20 years with LVH (LVMI in men > 120 g/m2; LVMI in women > 100 g/m2). Data were obtained from 411 of 505 randomized patients. INTERVENTIONS: Indapamide SR 1.5 mg, or enalapril 20 mg, daily for 48 weeks. MAIN OUTCOME MEASURES: LVMI variation in the perprotocol population. RESULTS: Indapamide SR 1.5 mg significantly reduced LVMI (-8.4 +/- 30.5 g/m2 from baseline; P< 0.001), but enalapril 20 mg did not (-1.9 +/- 28.3 g/m2). Indapamide SR 1.5 mg reduced LVMI significantly more than enalapril 20 mg: -6.5 g/m2, P = 0.013 (-4.3 g/m2 when adjusted for baseline values; P = 0.049). Both drugs equally and significantly reduced blood pressures (P< 0.001), without correlation with LVMI changes. Indapamide SR progressively reduced wall thicknesses throughout the 1-year treatment period. In contrast, the effect of enalapril observed at 6 months was not maintained at 12 months. CONCLUSIONS: Indapamide SR 1.5 mg was significantly more effective than enalapril 20 mg at reducing LVMI in hypertensive patients with LVH. PMID- 11057436 TI - Use of an age-adjusted Doppler E/A ratio in patients with moderate to severe hypertension. AB - OBJECTIVE: To assess the ratio of early (E) to late atrial (A) mitral Doppler peak flow velocity (Doppler E/A ratio) before and after adjustment for age in patients with moderate to severe hypertension, in whom left ventricular diastolic dysfunction is an early finding. Mitral flow patterns can be used to assess diastolic filling characteristics, and the Doppler E/A ratio is the parameter most commonly used, although it is known to be strongly age dependent. There are no established normal values for this ratio. DESIGN: Retrospective data analysis. SETTING: A 2000-bed tertiary-care teaching hospital. PATIENTS: We studied 190 patients (99 women and 91 men; ages 55 +/- 13 years) with moderate to severe hypertension. INTERVENTIONS: The ratio of early (E) to late atrial (A) mitral Doppler peak flow velocity was measured. As this ratio depends on age, a Z score was calculated to control for this influence. The Z score is the standardized normal deviation of the mean, with a normal value of 0 +/- 2. MAIN OUTCOME MEASURES: Sensitivities and specificities for detecting an age-dependent reduction in Doppler E/A score (Z score less than -2) with a non-age-dependent Doppler E/A ratio (less than 1) were calculated. RESULTS: In 106 of the patients (56%) the Doppler E/A ratio was less than 1.0. Only nine patients (4.7%) had a Z score less than -2. The sensitivity of the Doppler E/A ratio threshold of 1.0 for detecting a Z score less than -2 was 0.89 and the specificity was 0.46. A Z score less than -2 was found only in patients younger than 45 years. CONCLUSIONS: The Doppler E/A ratio was reduced in a large proportion of our patients. However, after correction for age it was decreased in only 4.7% of these patients. The use of a single Doppler E/A ratio threshold value has a weak diagnostic power to detect age-independent changes in mitral flow patterns. PMID- 11057437 TI - Haematocrit profoundly affects left ventricular diastolic filling as assessed by Doppler echocardiography. AB - BACKGROUND: The main determinants of diastolic function--pre- and afterload of the heart--are affected by the haematocrit, but the relation between haematocrit and diastolic function is unclear. OBJECTIVE: To study the association between interindividual haematocrit values and diastolic function, by echocardiography. DESIGN: In a cross-sectional survey, blood pressure, haematocrit values, and high quality Doppler indexes of left ventricular filling were obtained in 1297 individuals, 25-74 years of age, and analysed by regression analyses. RESULTS: Haematocrit and systolic blood pressure were strongly correlated (r = 0.23; P < 0.0001). Moreover, haematocrit was inversely correlated with the peak velocity of early left ventricular filling and with the peak velocity of early filling divided by late filling (E/A ratio; both P< 0.005). Left ventricular isovolumic relaxation time (IVRT) was positively associated with haematocrit (r= 0.18, P< 0.001). In individuals with an abnormal Doppler filling pattern (E/A(< 50 years) < 1, E/A(> 50 years) < 0.5, or IVRT(< 30 years) > 92 ms, IVRT30-50 years > 100 ms or IVRT> 50 years > 1 05 ms; n = 119), greater haematocrit values were observed than in those with normal diastolic parameters (P< 0.001). Conversely, individuals with an increased haematocrit (> 50% in men, > 45% in women; n = 16) had a greater risk of presenting with abnormal left ventricular filling (31.3%) compared with individuals with normal (12.1%; n = 898;) or low (< 40% in men, < 35% in women: 10.5%, n = 38; P = 0.07) haematocrit. Strong and significant associations between haematocrit and Doppler indexes of left ventricular filling were confirmed after adjustment for multiple potential confounders including blood pressure, antihypertensive medication and body mass index. Similarly, blood pressure and parameters of diastolic filling were strongly associated correlations that were not affected by inclusion of haematocrit values into the regression model. CONCLUSION: The data point to substantial adaptations of diastolic filling in response to both blood pressure and the characteristics of the medium that is propelled by the heart Therefore, in addition to blood pressure values, the variability of haematocrit values should be considered when diastolic function is being evaluated by Doppler echocardiography. PMID- 11057438 TI - Renal cyclic 3',5'-guanosine monophosphate and sodium excretion in Dahl salt resistant and Dahl salt-sensitive rats: comparison of the roles of bradykinin and nitric oxide. AB - OBJECTIVE: The purpose of this study was to determine the relative importance of bradykinin and nitric oxide (NO) in mediating renal responses to altered sodium intake in Dahl salt-resistant (Dahl-SR) and salt-sensitive (Dahl-SS) rats. DESIGN AND METHODS: Dahl-SR and Dahl-SS rats consumed a diet containing 0.15% (low) or 4.0% (high) sodium chloride for 10 days. A microdialysis technique was then used to measure renal cortical interstitial fluid (RIF) cyclic 3',5'-guanosine monophosphate (cGMP) production in anesthetized rats, under baseline conditions and during acute cortical infusion of either the bradykinin B2 receptor antagonist icatibant or the NO synthase inhibitor nitro-L-arginine methyl ester (L-NAME). Urine sodium excretion was monitored simultaneously by ureter cannulation. Results Baseline sodium excretion was similar in the two types of rats, but RIF cGMP was significantly elevated in Dahl-SR compared to Dahl-SS rats on both low and high sodium diets. Icatibant infusion significantly reduced both RIF cGMP and sodium excretion in Dahl-SR rats during low sodium intake, but had no effect in Dahl-SS rats on either diet L-NAME infusion significantly reduced sodium excretion in Dahl-SR and Dahl-SS rats, during both low and high sodium intake. L-NAME infusion caused a significant reduction in RIF cGMP in Dahl-SR and Dahl-SS rats on low sodium diet, but reduced RIF cGMP only in Dahl-SR rats on high sodium diet. Conclusion These data suggest a potential role for cortical bradykinin, but not NO, in mediating the differences in the renal response to low sodium intake between Dahl-SR and Dahl-SS rats. PMID- 11057439 TI - Tubulointerstitial injury and loss of nitric oxide synthases parallel the development of hypertension in the Dahl-SS rat. AB - OBJECTIVE: Alterations in renal nitric oxide (NO) are involved in the hypertension of the Dahl salt-sensitive (Dahl-SS) rat We sought to identify the kinetics and sites of expression of the major NO synthase (NOS) isoforms. DESIGN: The renal expression of the major NOS were examined in Dahl-SS and salt-resistant rats (Dahl-SR) while on a low salt (0.1% NaCl) diet at 3 and 9 weeks of age. METHODS: Renal biopsies from Dahl-SS and Dahl-SR rats were compared for evidence of renal injury and for alterations in expression of the NOS enzymes by quantitative immunohistochemistry. RESULTS: At 3 weeks of age Dahl-SS and Dahl-SR rats have normal renal histology and similar immunohistochemical expression of NOS1, -2, and -3. At 9 weeks Dahl-SS rats had significantly higher blood pressure than Dahl-SR rats (P< 0.005 ), and lower macula densa NOS1 (P< 0.05) and cortical and medullary NOS3 (P< 0.05). NOS2 was reduced in cortical tubules in biopsies showing severe tubulointerstitial damage, but was not significantly different between Dahl-SS and Dahl-SR groups as a whole. Dahl-SS rats also manifested glomerular and tubulointerstitial injury. Tubular expression of osteopontin (OPN), which is an inhibitor of NOS2, correlated with the systolic BP in individual Dahl-SS rats (r2 = 0.80, P < 0.0001 ). CONCLUSION: Tubulointerstitial injury and the loss of NOS occur after birth and parallel the development of hypertension. We suggest that the structural and functional changes that occur with renal injury in the Dahl-SS rat may contribute to the development of hypertension. PMID- 11057440 TI - Improvement of renal dysfunction in rats with chronic heart failure after myocardial infarction by treatment with the endothelin A receptor antagonist, LU 135252. AB - OBJECTIVE: To investigate the role of an activated endothelin system in the renal dysfunction observed in chronic heart failure after myocardial infarction. METHODS: In rats with heart failure after myocardial infarction and in sham operated animals (Sham), we investigated the effect on renal function of long term oral treatment with the selective endothelin A (ETA) receptor antagonist, LU 135252 (30 mg/kg per day; groups MI/LU and Sham/LU) or placebo (groups MI/P, Sham/P). Only animals with extensive myocardial infarction (at least 46% of the left ventricle) were included in the study. Infarct size was matched between groups MI/P and MI/LU. Endogenous creatinine clearance, fractional sodium excretion, and plasma and urinary concentrations of endothelin were determined 12 weeks after myocardial infarction. RESULTS: Endogenous creatinine clearance was significantly lower in group MI/P than in group Sham/P (MI/P: 0.64 +/- 0.05, Sham/P: 0.81 +/- 0.04 ml/min per 100 g body weight; P= 0.01 (means +/- SEM)). Treatment with LU 135252 completely prevented the decline in creatinine clearance in rats with chronic myocardial infarction (MI/LU: 0.98 +/- 0.21; Sham/LU: 0.83 +/- 0.10). Fractional sodium and protein excretion did not differ among the four groups. Group MI/P had a marked increase in plasma endothelin concentrations, which was not affected by treatment with LU 135252. Urinary endothelin excretion was significantly lower in group MI/P than in group Sham/P. In the treatment groups, no difference could be observed between animals that had suffered myocardial infarction and the sham-operated group, although LU 135252 markedly increased the urinary excretion of endothelin. CONCLUSION: Our data demonstrate a restoration of impaired renal function in chronic ischaemic heart failure by treatment with the selective ETA receptor antagonist, LU 135252. These results offer a promising therapeutic option for the treatment of renal insufficiency in patients with chronic heart failure. PMID- 11057441 TI - Haemodynamic and metabolic effects of rilmenidine in hypertensive patients with metabolic syndrome X. A double-blind parallel study versus amlodipine. AB - OBJECTIVE: To compare the effects of rilmenidine with those of amlodipine on blood pressure, glucose metabolism, plasma lipid concentration and fibrinolysis parameters. DESIGN: A four-month randomized double-blind, parallel group study. PATIENTS AND METHODS: Obese hypertensive patients with hypertriglyceridaemia (> or = 2.3 mmol/l) and impaired glucose tolerance (OMS-ADA) were included (n = 52). A placebo run-in period of 2 weeks was followed by 4 months of double-blind treatment with either rilmenidine or amlodipine. Blood pressure was recorded using a mercury sphygmomanometer. Glucose metabolism was evaluated by an oral glucose tolerance test RESULTS: Of the 52 patients recruited, 47 (21 rilmenidine and 26 amlodipine) completed the 4-month treatment period. The intention-to-treat analysis showed a comparable reduction in systolic and diastolic blood pressure (SBP, DBP) with the two anti-hypertensive treatments (rilmenidine -13.9/-13.5 mmHg; amlodipine - 17.6/-15.0 mmHg). Insulin concentrations under basal conditions and 2 h after a standard oral glucose load did not change significantly after treatment in both groups. Plasma glucose under basal conditions and 2 h after a standard oral glucose load as well as the area under the plasma glucose concentration curve tended to decrease in the rilmenidine group and to increase in the amlodipine group so that the changes in these parameters were significantly different between the two study groups (P= 0.041, P = 0.042 and P = 0.015, respectively). Plasminogen activator inhibitor type 1 (PAI 1) antigen and PAI-1 activity were only decreased in the rilmenidine group (not statistically significant). CONCLUSION: Our results demonstrate that rilmenidine and amlodipine have a comparable anti-hypertensive effect but only rilmenidine is able to improve glucose metabolism. PMID- 11057442 TI - High-normal serum homocysteine concentrations are associated with an increased risk of early atherosclerotic carotid artery wall lesions. PMID- 11057443 TI - Functional analysis of the leader peptide of the yeast gene CPA1 and heterologous regulation by other fungal peptides. AB - The 25-amino-acid leader peptide present at the 5' end of yeast CPA1 mRNA is responsible for the translational repression of that gene by arginine. We show here that the active domain of the yeast peptide is highly specific and extends over amino acids 6-23. The region between amino acids 6-21 is well conserved between similar peptides present upstream of CPA1-homologous genes in other fungi. The Neurospora crassa arg-2 peptide represses the expression of CPA1, whereas the peptide from Aspergillus nidulans has only a weak regulatory effect. Such results suggest that the N- and C-terminal amino acids of the peptide may influence its regulatory activity. We also show that the transcription start sites of CPA1 are not modified when the cells are grown in the presence of arginine, nor in a strain carrying an inactive peptide. PMID- 11057444 TI - The genetic control of spontaneous and UV-induced mitotic intrachromosomal recombination in the fission yeast Schizosaccharomyces pombe. AB - An artificially created non-tandem hetero-allelic duplication was constructed to assay mitotic intrachromosomal recombination in Schizosaccharomyces pombe. Two classes of recombinants could be distinguished: deletion-types, in which one copy of the duplicated sequence and the intervening sequence were lost, and conversion types which retained the duplication. For spontaneous recombination, compared to wild-type cells, a rad22 mutant (corresponding to a Saccharomyces cerevisiae rad52 mutant) had wild-type levels of deletion-types, but was hypo-recombinant for conversion-types; rad16 (S. cerevisiae rad1), rad22 rad16 (S. cerevisiae rad52 rad1) and swi10 (S. cerevisiae rad10) mutants were hyper-recombinant for both types; rad22 swi10 (S. cerevisiae rad52 rad10) mutants were hypo-recombinant for both types; rhp51 (S. cerevisiae rad51) and rhp54 (S. cerevisiae rad54) mutants were hyper-recombinant for deletion-types, but almost completely lacked conversion-types. For wild-type cells, UV-irradiation induced both types of recombinant, but mainly conversion-types. All of the mutants lacked UV-induced recombination. PMID- 11057445 TI - Sulfate assimilation in Aspergillus terreus: analysis of genes encoding ATP sulfurylase and PAPS-reductase. AB - Two genes for the sulfate assimilation pathway in Aspergillus terreus were cloned. The genes sAT (coding for PAPS-reductase) and sCT (coding for ATP sulfurylase) form a small gene cluster. Both genes are similar to their homologs in A. nidulans (sA and sC), Penicillium chrysogenum (aps) and Saccharomyces cerevisiae (MET3 and MET16). In the coding sequence of the sCT gene, a typical non-functional APS-kinase-like domain is present. The sCT gene is expressed in A. nidulans, but its expression there is less sensitive to methionine level than in the original species. Two regions 5' upstream of sAT were found to be similar to those of sA. PMID- 11057446 TI - Yeti--a degenerate gypsy-like LTR retrotransposon in the filamentous ascomycete Podospora anserina. AB - In the filamentous ascomycete Podospora anserina a 6,935-bp retrotransposon, Yeti, has been identified and characterized. It is flanked by a 5-bp target site duplication and contains long terminal repeats (LTRs) 354 bp in length. The LTRs show a high degree of identity to the previously reported repetitive element repa, a sequence suggested to represent a solo-LTR element of an unknown transposon. In the investigated Podospora strains, the number of complete Yeti copies is significantly lower than the number of repa elements, with up to 25 copies. Yeti appears to be inactive: it is highly degenerate and no transcripts of the element have been detected even in Podospora cultures grown under elevated stress conditions. The amino acid sequences deduced from Yeti display significant homology, particularly in the reverse transcriptase region, to those of other fungal retrotransposons, indicating that it is a member of the gypsy family. As suggested by the unusual dinucleotide content, degeneration of Yeti appears to be the result of a molecular mechanism resembling repeat-induced point mutation in Neurospora crassa. PMID- 11057447 TI - Telomere-associated RFLPs and electrophoretic karyotyping reveal lineage relationships among race-specific strains of Ustilago hordei. AB - The inheritance of telomere-associated restriction fragment length polymorphisms (tel-RFLP) and chromosome-length polymorphisms (CLPs) were criteria used for the identification of strains of Ustilago hordei that form a direct lineage. Teliospore collections of race 8 strains and strains reported to be derived from race 8 through inbreeding were used in these analyses. None of the race 8 strains examined in this study, representing three consecutive inbred generations, was polymorphic for any terminal BamHI and BglII chromosomal loci, nor did they have any apparent CLPs. Strains from a teliospore collection obtained in 1971 and designated 447, representing the first inbred generation of race 8 strains and a shift to increased virulence on cultivar Hannchen, had tel-RFLP arrays indistinguishable from the race 8 strains isolated in this study; and they had no obvious CLPs. Strains from the presumed second inbred generation, the 1279 teliospore line, which was pathogenic on six additional cultivars, had numerous CLPs and a tel-RFLP array that differed from race 8 strains at more than 50% of the terminal chromosomal BamHI and BglII restriction sites. The tel-RFLP array of each 1279 strain was identical and indistinguishable from the arrays of strains representing races 10 and 13, indicating that they share a direct lineage. A race 14 strain, also presumed to be derived from race 8 strains by inbreeding, had a unique tel-RFLP array and an electrophoretic karyotype that distinguished it from all other strains. The tel-RFLP arrays alone eliminate the 1279 and race 14 strains from being direct descendants from race 8 strains by inbreeding and suggest that this approach can identify a strain lineage among other inbred sexually reproducing fungi, or isolates that comprise different asexual clonal populations. PMID- 11057448 TI - Characterization of laboratory mutants of Venturia inaequalis resistant to the strobilurin-related fungicide kresoxim-methyl. AB - Several agricultural fungicides related to the antifungal strobilurins act as inhibitors of respiration by binding to mitochondrial cytochrome b. Two types of laboratory mutants resisting higher doses of the strobilurin-related inhibitor kresoxim-methyl were characterized for Venturia inaequalis, the causal agent of apple scab. Selection of mutagenized conidia by kresoxim-methyl yielded mutants altered in the expression of alternative respiration during the stage of conidia germination. Cytochrome b sequences were not affected in the respective mutants. Selection of conidia on media containing the alternative oxidase inhibitor salicylhydroxamic acid in addition to kresoxim-methyl yielded a highly resistant mutant distinguished by a G143A exchange in cytochrome b. The status of mitochondrial cytochrome b genes remained heteroplasmic, and mitochondria containing wild-type cytochrome b returned to high frequencies during cultivation on inhibitor-free medium. However, continuation of the selection process led to a more pronounced replacement of sensitive by mutated mitochondria. The G143A mutation of cytochrome b causing resistance of V. inaequalis to a strobilurin related inhibitor has been reported previously for mouse mitochondria; and a permanent G143A exchange rendering naturally resistant mitochondria has been reported for the strobilurin-producing basidiomycete Mycena galopoda and for the sea urchin Paracentrotus lividus. At the corresponding position, alanine was also present in chloroplast cytochrome b6 exhibiting low binding of strobilurin related inhibitors. The mutation of cytochrome b reported here for V. inaequalis describes the first example of a mutation in filamentous ascomycetes and is part of an assessment of resistance risks inherent to strobilurin fungicides. PMID- 11057449 TI - Nuclear control of mitochondrial genome reorganization characterized using cultured cells of ditelosomic and nullisomic-tetrasomic wheat lines. AB - In vitro-mediated reorganization of the mitochondrial genome is governed by information contained in the nuclear genome. Here we show, from the study of tissue cultures initiated from an almost complete collection of ditelosomic and nullisomic-tetrasomic wheat lines, that nuclear control of the mitochondrial genome structure is a highly complex process. Whereas information responsible for the amplification of defined molecular configurations has been found to be located in either a single or a few chromosomal arms, other rearrangements such as changes in the relative amounts of interconverting subgenomic structures are governed by a number of genes scattered over most of the wheat chromosomes. PMID- 11057450 TI - Beneficial microbes: health or hazard? AB - Normal microbial flora support the health of the host by diverse mechanisms. When antibiotics, stress, disease or medications disrupt normal microflora, the ability to ward off infection by pathogens is compromised. The use of beneficial microbes (also known as biotherapeutic agents, probiotics, synbiotics) has been shown to be an effective therapeutic agent for some diseases. Various types of diarrhoea (antibiotic-associated diarrhoea, Clostridium difficile disease, traveller's diarrhoea) are most responsive to these beneficial microbes. Serious risks associated with these microbes are largely theoretical at this point, but the risks need to be studied as the use of these beneficial microbes increases in popularity. Beneficial microbes are living organisms used as therapeutic agents to restore the health of the host in times when normal microflora have been disturbed. The efficacy to prevent or treat diarrhoea has been documented in multiple large, placebo-controlled, blinded clinical trials with only a few of these beneficial microbes. Risks of these beneficial microbes are limited, but potential risks have not been extensively studied in large numbers of patients. PMID- 11057451 TI - Gastrostomy placement in patients with Crohn's disease. AB - Crohn's disease often involves the stomach, yet a permanent enterocutaneous fistula does not usually occur, after a percutaneous endoscopic gastrostomy is removed from a patient with Crohn's disease. This is because the factors that are related to the non-closure of a fistula are absent or have been treated (distal bowel obstruction, abdominal sepsis, undernutrition, poor gastric blood supply or abnormal serum levels of C-reactive protein and albumin). Gastric involvement in patients with Crohn's disease is common. Enterocutaneous fistulas from the stomach are rare. Percutaneous endoscopic gastrostomy (PEG) insertion and subsequent removal rarely cause problems in patients with Crohn's disease. Endoscopic removal of a PEG is advised in patients with Crohn's disease. Steroids may delay gastric adhesion to the anterior abdominal wall. PMID- 11057452 TI - Micro-organisms administered in the benefit of the host: myths and facts. AB - OBJECTIVE: To evaluate the published literature on the potential benefit of micro organisms on the general well being of the host. STUDY DESIGN: All published prospective, randomized, placebo-controlled trials with micro-organisms to improve the health of the host were critically reviewed. RESULTS: According to published data, there is evidence suggesting that Lactobacillus rhamnosus strain GG or Lactobacillus casei strain GG and Saccharomyces boulardii may be of possible benefit for the treatment of several medical conditions. However, published data on the therapeutic effect of other micro-organisms are almost non existent. CONCLUSION: Better designed prospective, randomized, and placebo controlled studies are needed. Most of the present strains have not been selected in a rational way, but apparently represent rather randomly picked isolates. Although the theoretical advantages of micro-organisms administered to the benefit of the host are extremely interesting and promising, results of clinical trials are disappointing. PMID- 11057453 TI - The safe use of percutaneous gastrostomy for enteral nutrition in patients with Crohn's disease. AB - OBJECTIVE: To determine the safety of percutaneous endoscopic gastrostomy (PEG) tube placement for nutritional support and/or defined therapeutic enteral nutrition (TEN) in adult patients with Crohn's disease. DESIGN: A prospective, observational study of patients with Crohn's disease in whom PEG tubes were placed for nutritional support or TEN. SETTING: A specialist nutrition clinic at a gastroenterology tertiary referral centre in Harrow, UK. PARTICIPANTS: Nine patients with Crohn's disease. Seven patients had nutritional failure and were unable to tolerate nasogastric feeding, and two patients were recruited in whom TEN therapy for active disease was indicated. The age range was 21-52 years (median, 30 years). Five patients were female; all had had previous ileo-colonic resections, one had a gastro-enterostomy and one had a non-healing Crohn's related gastric ulcer. INTERVENTIONS: PEG insertion (Fresenius, Frecka 9 Fr) was performed at endoscopy with intravenous sedation. Follow-up with tubes in situ was for a median of 37 weeks (range, 4-276 weeks), and for a further median of 80 weeks (range, 52-120 weeks) in those whose tubes have been removed. MAIN OUTCOME MEASURES: The level of disease activity, nutritional status/body mass index and any complications associated with PEG tube placement were recorded. RESULTS: PEG was achieved in all patients; the only complication was a minor superficial entry site infection. Five patients continue to use PEG feeding to good effect, including healing of the Crohn's-associated ulcer. One patient now eats normally having regained target weight, and three require parenteral nutrition, having failed to achieve nutritional sufficiency despite an optimal enteral regimen via the PEG. An adverse body image in one of these patients (an opiate abuser with a long psychiatric history) was probably contributory to PEG failure. There was no peristomal or fistulous disease. CONCLUSIONS: Although nutrition via PEG is not always successful, failures are of enteral nutrition, and not of the means. PEG use in selected patients with Crohn's disease appears safe and can prove a useful addition to therapeutic options. PMID- 11057454 TI - Predicting early mortality following percutaneous stent insertion for malignant biliary obstruction: a multivariate risk factor analysis. AB - BACKGROUND: Percutaneous stent placement is an accepted method of palliation in malignant biliary obstruction. Factors predicting early mortality after this procedure have not been identified. METHODS: We performed a retrospective study of 141 patients with malignant biliary obstruction who underwent percutaneous stent placement for biliary decompression to identify the risk factors associated with early mortality (< or = 30 days). RESULTS: Of 14 clinicopathological and laboratory variables analysed blood urea, albumin, haemoglobin and alkaline phosphatase were found to be significant on univariate analysis. The age and gender of the patient along with cancer type, level of obstruction, presence of pyrexia and bilirubin level had no influence on early mortality. Stepwise logistic regression identified the haemoglobin level and blood urea to be independently significant in predicting early mortality. Overall 30-day mortality was 20.5% (29/141). Patients with blood urea over 4.3 mmol/l and a haemoglobin less than 10.9 g/dl had a mortality rate of 52% (12/23) compared with 14% (17/118) in the remainder. Using these two variables a regression equation has been derived which allows calculation of the probability of survival at 30 days after the percutaneous procedure. CONCLUSIONS: Laboratory variables in patients with malignant obstructive jaundice can be used to predict mortality following percutaneous stent insertion. PMID- 11057455 TI - A cross-sectional and longitudinal study of health-related quality of life after percutaneous gastrostomy. AB - OBJECTIVE: Although gastrostomy feeding tends to have fewer interruptions than naso-gastric feeding and is cosmetically more acceptable; there is little information on health-related quality of life (HRQoL) in these patients. This study aimed to measure the health-related quality of life of patients after gastrostomy placement. DESIGN: Cross-sectional and prospective cohorts. SETTING: Institutional and community-dwelling patients receiving nutritional support via a gastrostomy. PARTICIPANTS: For the cross-sectional cohort, all individuals who have received a percutaneous gastrostomy from our unit (January 1994-December 1996) were included; 55 of the 102 patients who were still living agreed to follow-up assessment. For the prospective cohort, all patients referred to our unit for percutaneous gastrostomy (March 1997-June 1998) were eligible to participate; 54 of 88 patients (62%) consented and were recruited. METHODS AND OUTCOME MEASURES: A cross-sectional assessment of patient outcome and health related quality of life using SF-36, the Hospital Anxiety and Depression Scale and a disease-specific questionnaire (PEG-Qu) was carried out in patients in primary health care after receiving a gastrostomy. These assessments, plus the Modified Rankin Scale, were performed on patients studied at the time of gastrostomy placement, and after 1, 6 and 12 months of follow-up. RESULTS: HRQoL questionnaires could be answered in less than half the patients. An overall rating of the effect of the gastrostomy on the patients' and carers' HRQoL showed a positive effect in 55% and 80%, respectively. A positive impression of the gastrostomy by the patient did not necessarily reflect an improvement in their nutritional status. Assessment of HRQoL in a cohort of gastrostomy patients showed deficiencies in the physical domain but not mental function (anxiety or depression), energy or health perception. Neither physical function nor level of cognition at time of gastrostomy placement appear to be able to predict patient survival. Nutritional outcome was not related to HRQoL outcome. CONCLUSION: The majority of patients and carers rate gastrostomy positively. Patients who were 75 years or older had a poorer survival compared to younger patients, but gender, physical or cognitive function had no predictive value on survival. PMID- 11057456 TI - Endoscopic ligation of oesophageal varices compared with injection sclerotherapy in primary biliary cirrhosis. AB - BACKGROUND AND AIMS: Oesophageal varices are an important complication in primary biliary cirrhosis (PBC). However, there have yet to be any studies made on treatment of oesophageal varices in PBC. We therefore studied the efficacy and related complications of endoscopic variceal ligation (EVL) and endoscopic injection sclerotherapy (EIS) as an initial treatment in primary biliary cirrhotic patients. METHODS: From December 1985 to March 1999, 29 biliary cirrhotic Japanese patients with portal hypertension and oesophageal varices were treated in our clinics. Eleven patients were treated with EVL and EIS, and 18 patients underwent EIS only. The liver function, renal function and respiratory function were studied before and after endoscopic treatment and any complications were also examined. RESULTS: In stages III and IV, significant differences were observed in the serum levels for total bilirubin and gamma-glutamic pyruvic transaminase only in the EIS group. Significant differences were observed in the rate of appearance of pyrexia, retrosternal pain and pleural effusion between the EIS and EVL groups. CONCLUSION: EVL significantly reduced the adverse effects associated with EIS at the initial session in primary biliary cirrhotic patients. PMID- 11057457 TI - Evaluation of a commercial serological kit for detection of salivary immunoglobulin G to Helicobacter pylori: a multicentre study. AB - OBJECTIVE: To evaluate the usefulness of a commercial serological enzyme immunoassay (EIA) for detecting salivary immunoglobulin (Ig) G to Helicobacter pylori. DESIGN: Prospective, multicentre study. SETTING: The study was conducted at 11 gastroenterology hospital centres throughout Italy. PARTICIPANTS AND INTERVENTIONS: Two hundred and thirteen dyspeptic patients underwent gastroscopy with antral biopsies. At each centre, two of the following three tests for H. pylori diagnosis were performed: urease quick test, histology, and 13C-urea breath test. Samples of unstimulated saliva and venous blood were collected from each patient. Salivary and serum H. pylori IgG were determined with the EIA Helori-test IgG (Eurospital, Trieste, Italy). MAIN OUTCOME MEASURES: Sensitivity, specificity, positive and negative predictive values, and accuracy of the salivary and serum EIAs. RESULTS: By using a receiver operating characteristic curve, salivary EIA yielded 81% (95% confidence interval 73-87%) sensitivity, 73% (62-83%) specificity, 84% (76-90%) positive predictive value, 69% (58-79%) negative predictive value, and 78% (72-84%) accuracy. At the cut-off suggested by the manufacturer, serum EIA had 90% (84-95%) sensitivity, 78% (67-86%) specificity, 88% (81-93%) positive predictive value, 82% (71-90%) negative predictive value, and 86% (81-91%) accuracy. CONCLUSION: In this large multicentre study, detection of salivary H. pylori IgG with a commercial serological EIA was a fairly accurate diagnostic method. Data confirm that saliva testing does not compare favourably with serology in the assessment of H. pylori status. PMID- 11057458 TI - Serum hyaluronan--a non-invasive test for diagnosing liver cirrhosis. AB - INTRODUCTION: Hyaluronan is a glucosaminoglycan synthesized by the mesenchymal cells and degraded by hepatic sinusoidal endothelial cells by a specific receptor mediated process. Elevated levels are associated with the sinusoidal capillarization that is seen in cirrhosis. METHODOLOGY: Serum hyaluronan was measured, using a radiometric assay (Pharmacia, Sweden) in 221 patients with biopsy-proven chronic liver disease of a variety of aetiologies (alcohol n = 70, autoimmune chronic active hepatitis n = 23, primary biliary cirrhosis n = 17, hepatitis C n = 69, cryptogenic n = 15, various n = 27). All patients were fasted, and their liver function tests, full blood count, prothrombin time and Child-Pugh score were assessed at the time of the liver biopsy. RESULTS: Hyaluronan levels (microg/l) were significantly higher in patients with liver cirrhosis (cirrhosis n = 127, mean 440, 95% CI 367-515) (P < 0.0001) compared with hepatic fibrosis (n = 23, mean 144, 95% CI 69-190), chronic hepatitis (n = 60, mean 63, 95% CI 37-91) and fatty liver (n = 11, mean 107, 95% CI 37-177). Within the cirrhotic population, there was no significant difference in hyaluronan levels between different aetiologies, but hyaluronan level increased proportionally to the severity of cirrhosis. Overall, a hyaluronan level > 100 microg/l had a 78% specificity and 83% sensitivity for diagnosing cirrhosis, while the specificity was increased to 96% for all patients with hyaluronan levels > 300 microg/l. The highest specificity and sensitivity were seen at a cut off value of 100 microg/l in patients with alcohol-associated liver disease (89%, 87%) and hepatitis C (93%, 72%) respectively. Within patient cohorts, there was a significant correlation (P < 0.01) between hyaluronan and albumin, platelet count and bilirubin, but not with alanine aminotransferase. CONCLUSION: Measurement of fasted serum hyaluronan reliably differentiated cirrhotic from non-cirrhotic liver disease and can be regarded as a useful test in the diagnosis of liver cirrhosis, particularly when a liver biopsy is contraindicated. PMID- 11057459 TI - Isolation of a single strain of Helicobacter pylori from the antrum and body of individual patients. AB - OBJECTIVES: This study aimed to determine intra-patient colonization patterns of Helicobacter pylori strains based on DNA fingerprinting and antibiotic susceptibility. METHODS: Two biopsies, one from the antrum and one from the body of the stomach, were taken from 97 patients. Prior informed consent was obtained. The status of cagA gene of H. pylori strains was analysed by using the polymerase chain reaction (PCR) technique, while DNA fingerprints were generated by PCR based, random amplified polymorphic DNA (RAPD) fingerprinting. The antibiotic susceptibility of the H. pylori isolates was examined by the disk diffusion method. RESULTS: A total of 51 pairs of H. pylori strains were isolated from both antrum and body specimens of 51 patients. This included two patients who were endoscoped twice because of treatment failure. All strains were positive for cagA gene by PCR. These 51 patients were found to harbour a single strain of H. pylori with identical or highly similar DNA profiles by PCR-based RAPD fingerprinting. In four of the 51 pairs, the DNA patterns of H. pylori from antrum and body showed minor differences, while three pairs of strains with different metronidazole sensitivities showed identical DNA fingerprints. Interestingly, the two treatment failure patients remained colonized with the strains that had the same RAPD fingerprinting patterns before and after treatment. CONCLUSION: The present study demonstrates that a single H. pylori strain colonizes a single stomach. However, this single genotypic strain may exhibit different metronidazole susceptibility in different parts of stomach. PMID- 11057460 TI - Granulocyte colony-stimulating factor improves function of neutrophils from patients with acute liver failure. AB - OBJECTIVE: To evaluate the in vitro effects of granulocyte colony-stimulating factor (G-CSF) on function of neutrophils in acute liver failure (ALF). METHODS: Neutrophil functions (superoxide and hydrogen peroxide production; phagocytosis and killing; complement receptor expression) were determined simultaneously in 23 patients with ALF due to paracetamol overdose and compared with 23 healthy control subjects. RESULTS: Phagocytosis was reduced in neutrophils from ALF patients compared to controls (P< 0.005) and was significantly increased by incubation with 1,000 or 5,000 IU/ ml G-CSF (P< 0.05). This correlated with increased expression of CD11b (r= 0.93) and CD18 (r= 0.98) after incubation with 5,000 IU/ml G-CSF (P< 0.05). Killing was reduced in ALF neutrophils compared to controls (P< 0.005) and was similarly restored by G-CSF (P< 0.005). An increase in killing correlated with increases in production of superoxide (r = 0.96) and hydrogen peroxide (r= 0.97) by ALF neutrophils after incubation with 1,000 and 5,000 IU/ml of G-CSF when formylmethionylleucylphenylalanine (fMLP) was the stimulant. G-CSF at 5,000 IU/ml increased the production of hydrogen peroxide (P< 0.01) when zymosan was the stimulant. CONCLUSIONS: G-CSF improves the neutrophil dysfunction of ALF. PMID- 11057461 TI - Thrombolytic therapy in patients with portal vein thrombosis: case report and review of the literature. AB - A 29-year-old male patient with Crohn's disease of the terminal ileum and previous abdominal surgery was admitted because of severe abdominal pain and signs of bacterial sepsis. The diagnosis of portal vein thrombosis and multiple liver abscesses due to Streptococcus intermedius septicaemia was made and antibiotic therapy was instituted immediately. As high-dose heparin therapy was ineffective, urokinase was administered intravenously over a total of 7 days. Within 2 days, the patient's symptoms completely subsided. Colour duplex ultrasonography revealed complete recanalization of the main stem of the portal vein; the right branch of the portal vein, however, remained occluded. Other case reports on thrombolytic therapy in patients with portal vein thrombosis are reviewed. PMID- 11057462 TI - Incidental adult nesidioblastosis after distal pancreatectomy for endocrine microadenoma. AB - Persistent hyperinsulinaemic hypoglycaemia (nesidioblastosis) presenting with hypoglycaemia is extremely rare in adults. The features are suggestive of an insulinoma with a vague presentation and delayed diagnosis. We describe a report of adult nesidioblastosis in association with a pancreatic endocrine microadenoma. PMID- 11057463 TI - Dissecting intramural haematoma of the oesophagus. AB - Dissecting intramural haematoma of the oesophagus is an under-recognized cause of sudden onset chest pain. Diagnosis is aided by the common co-existence of haematemesis, odynophagia, and dysphagia. Oesophagoscopy, barium swallow and cross-sectional radiology are all appropriate modalities of investigation. The latter is useful in excluding aortic dissection from the differential diagnosis. The condition usually follows a benign course with spontaneous healing requiring supportive treatment only. A systematic analysis of all the cases of dissecting intramural haematoma of the oesophagus (DIHO) reported in the worldwide literature reveals that this is essentially a benign condition. Eighty per cent of patients have at least two of the three typical presenting features of chest pain, haematemesis and dysphagia or odynophagia. Forty-nine per cent of cases are associated with sudden pressure changes within the oesophagus (e.g. due to swallowing) or secondary to direct trauma to the oesophagus. A further group appears to arise spontaneously and may be associated with underlying abnormal pressure changes within the oesophagus or a bleeding tendency. Awareness of the condition prevents the pain being mistakenly attributed to a cardiac cause. With conservative management the symptoms usually resolve within 2 weeks of presentation. The only mortality associated with DIHO is due to operative intervention or where there is another underlying life-threatening condition. PMID- 11057464 TI - The chevron osteotomy for correction of hallux valgus. Comparison of findings after two and five years of follow-up. AB - BACKGROUND: The chevron osteotomy, an accepted method for the correction of mild and moderate hallux valgus, is generally advocated for patients younger than the age of fifty years. The purposes of this prospective study were to compare the short-term (two-year) and intermediate-term (five-year) results of this operation with respect to patient satisfaction, flexion and extension of the metatarsophalangeal joint, maintenance of correction, and development of arthrosis and to determine whether the effectiveness of the procedure was limited by age. METHODS: Between April 1991 and September 1992, the chevron osteotomy was performed for the treatment of mild-to-moderate hallux valgus deformity in sixty six consecutive feet. Forty-three patients (fifty-seven feet) were available for follow-up at both two and five years postoperatively. The two-year and five-year clinical assessments were based on the American Orthopaedic Foot and Ankle Society's hallux-metatarsophalangeal-interphalangeal scale. RESULTS: Between the two-year and five-year follow-up evaluations, there was only a minimal change in overall patient satisfaction, and the average score on the hallux metatarsophalangeal-interphalangeal scale was unchanged. The passive range of motion of the first metatarsophalangeal joint decreased between the preoperative assessment and the two-year follow-up evaluation and was unchanged at the five year follow-up evaluation. Radiographic evaluation showed no changes in the hallux valgus or intermetatarsal angle between the two-year and five-year evaluations, although the number of feet with arthrosis of the metatarsophalangeal joint increased slightly, from eight to eleven. Patients aged fifty years or older did as well as younger patients. CONCLUSIONS: At these two follow-up periods, the chevron osteotomy was found to be a reliable procedure for the correction of mild and moderate hallux valgus deformity, and outcome did not differ on the basis of age. PMID- 11057465 TI - Functional outcome of semiconstrained total elbow arthroplasty. AB - BACKGROUND: The objective of the present study was to review the results of primary total elbow arthroplasty with use of the Coonrad-Morrey prosthesis. Two hypotheses were tested: (1) the results in patients with inflammatory arthritis would be superior to those in patients with a traumatic or posttraumatic condition, and (2) the isometric extensor torque after total elbow arthroplasty would be significantly less than that of the contralateral elbow. METHODS: Forty seven consecutive patients (fifty-one elbows) had the operation performed by one of three surgeons between November 1, 1989, and June 30, 1996. Thirty-six surviving patients (thirty-nine elbows) were available for follow-up. The mean duration (and standard deviation) of follow-up was 50 +/- 11 months (range, twenty-four to ninety-seven months). The mean age at the time of the operation was 64 +/- 11 years (range, twenty-seven to eighty-seven years). Eighteen patients (twenty-one elbows) had inflammatory arthritis. Eighteen patients (eighteen elbows) had an acute fracture or posttraumatic condition (posttraumatic osteoarthritis in eight, an acute fracture of the humerus in seven, nonunion of the distal aspect of the humerus in two, and primary osteoarthritis in one). The patients were evaluated with use of questionnaires (the Mayo elbow performance index, the Short Form-36 [SF-36], and the Disabilities of the Arm, Shoulder and Hand [DASH] Questionnaire); clinical examination by an orthopaedic surgeon who was not involved with the pre-operative, operative, postoperative, or follow-up care; radiographs; and elbow strength-testing with an isokinetic dynamometer. RESULTS: The mean score (and standard deviation) on the Mayo elbow performance index for the group that had inflammatory arthritis (90 +/- 11 points) was significantly higher than that for the group with a traumatic or posttraumatic condition (78 +/- 18 points) at the time of the latest follow-up (p < 0.05). In both groups, the mean extensor torque of the involved elbow was significantly less than that of the contralateral elbow (p < 0.05). No significant difference between the groups was found with respect to the flexion-extension arc of motion. Ten elbows (26 percent) had ulnar nerve dysfunction (a transient deficit in six and a permanent deficit in four); nine (23 percent), an intraoperative fracture (of the humeral diaphysis in four, of the ulnar diaphysis in four, and of the olecranon in one); three (8 percent), a periprosthetic infection; three, a triceps disruption; and one (3 percent), a revision because of a fracture of the ulnar component. There were no other revisions. Of the thirty-four elbows with complete radiographic follow-up, twenty-three had no change in the bone-cement interface. Progressive radiolucency was noted around the ulnar prosthesis in eight elbows, around the humeral prosthesis in one elbow, and around both components in two elbows. CONCLUSIONS: Patients who had a total elbow arthroplasty with use of a semiconstrained Coonrad-Morrey prosthesis were generally satisfied; the mean level of patient satisfaction was 9.2 of a possible 10 points for those who had inflammatory arthritis and 8.6 points for those who had a fracture or posttraumatic condition. The rates of complications involving the ulnar nerve, intraoperative fracture, triceps disruption, deep infection, and periprosthetic radiolucency are of concern. PMID- 11057466 TI - Histological changes in the human anterior cruciate ligament after rupture. AB - BACKGROUND: Four phases in the response to injury of the ruptured human anterior cruciate ligament are observed histologically; these include an inflammatory phase, an epiligamentous repair phase, a proliferative phase, and a remodeling phase. One objective of this study was to describe the histological changes that occur in the ruptured human anterior cruciate ligament during these phases. Myofibroblast-like cells that contain alpha-smooth muscle actin are present in the midsubstance of the intact human anterior cruciate ligament. A second objective of this study was to determine whether an increased number of myofibroblast-like cells is found in the midsubstance of the ruptured human anterior cruciate ligament because it was thought that those cells might be responsible in part for the retraction of the ruptured anterior cruciate ligament. In the early phase of this study, it was found that the number of myofibroblast-like cells in the midsubstance of the ruptured anterior cruciate ligament was actually decreased, and this hypothesis was abandoned. During the epiligamentous repair phase, synovial tissue was formed that covered the ends of the ruptured anterior cruciate ligament. Most of the synovial lining cells were myofibroblast-like cells that contained alpha-smooth muscle actin. The primary objective of this study was to determine the location and the characteristics of the alpha-smooth muscle actin-containing myofibroblast-like cells that appear in the human anterior cruciate ligament following rupture. METHODS: Twenty-three ruptured and ten intact human anterior cruciate ligaments were evaluated for cellularity, nuclear morphology, blood vessel density, and percentage of cells containing a contractile actin isoform, alpha-smooth muscle actin. The histological features of the synovial and epiligamentous tissues were also described. RESULTS: At no time after rupture was there evidence of tissue bridging between the femoral and tibial remnants of the anterior cruciate ligament. The ruptured ligaments demonstrated a time-dependent histological response, which consisted of inflammatory cell infiltration up to three weeks, gradual epiligamentous repair and resynovialization between three and eight weeks, and neovascularization and an increase in cell number density between eight and twenty weeks. Compared with the intact ligaments, there was a decrease in the percentage of myofibroblast-like cells containing alpha-smooth muscle actin within the remnant of the ligament. However, many of the epiligamentous and synovial cells encapsulating the remnants contained alpha-smooth muscle actin. CONCLUSIONS: After rupture, the human anterior cruciate ligament undergoes four histological phases, consisting of inflammation, epiligamentous regeneration, proliferation, and remodeling. The response to injury is similar to that reported in other dense connective tissues, with three exceptions: formation of an alpha smooth muscle actin-expressing synovial cell layer on the surface of the ruptured ends, the lack of any tissue bridging the rupture site, and the presence of an epiligamentous reparative phase that lasts eight to twelve weeks. Other characteristics reported in healing dense connective tissue, such as fibroblast proliferation, expression of alpha-smooth muscle actin, and revascularization, also occur in the ruptured human anterior cruciate ligament. CLINICAL RELEVANCE: Unlike extra-articular ligaments that heal after injury, the human intra articular anterior cruciate ligament forms a layer of synovial tissue over the ruptured surface, which may impede repair of the ligament. Moreover, a large number of cells in this synovial layer and in the epiligamentous tissue express the gene for a contractile actin isoform, alpha-smooth muscle actin, thus differentiating into myofibroblasts. These events may play a role in the retraction and lack of healing of the ruptured anterior cruciate ligament. PMID- 11057467 TI - Idiopathic adhesive capsulitis. A prospective functional outcome study of nonoperative treatment. AB - BACKGROUND: Idiopathic adhesive capsulitis is a commonly recognized but poorly understood cause of a painful and stiff shoulder. Although most orthopaedic literature supports treatment with physical therapy and stretching exercises, some studies have demonstrated late pain and functional deficits. The purpose of this study was to evaluate the outcome of patients with idiopathic adhesive capsulitis who were treated with a stretching-exercise program. METHODS: Seventy five consecutive patients (seventy-seven shoulders) with phase-II idiopathic adhesive capsulitis were treated with use of a specific four-direction shoulder stretching exercise program and evaluated prospectively. The initial evaluation included the recording of a detailed medical and orthopaedic history and assessment of pain, range of motion, and function. The outcome evaluation included assessment of pain, range of motion, and function; completion of the Disabilities of the Arm, Shoulder, and Hand (DASH) Questionnaire; and completion of the Short Form-36 (SF-36) Health Survey. The mean duration of follow-up was twenty-two months (range, twelve to forty-one months). One patient died prior to the final evaluation, and three patients were lost to follow-up. RESULTS: Sixty four (90 percent) of the patients reported a satisfactory outcome. Seven (10 percent) were not satisfied with the outcome, and five (7 percent) underwent manipulation and/or arthroscopic capsular release. The outcomes of the patients who did not have manipulation or capsular release were evaluated. There were significant improvements in the scores for pain at rest (from a mean of 1.57 points before treatment to a mean of 1.16 points at the final evaluation; p < 0.001) and pain with activity (from a mean of 4.12 points before treatment to a mean of 1.33 points at the final evaluation; p < 0.0001). On the average, active forward elevation increased 43 degrees, active external rotation increased 25 degrees, passive internal rotation increased eight vertebral levels, and the glenohumeral rotation arc at 90 degrees of abduction increased 72 degrees (p < 0.00001). The number of "yes" responses to the Simple Shoulder Test increased from a mean of 4.1 (of a possible twelve) to a mean of 10.75 (p < 0.00001). Despite the significant improvements and the high rate of patient satisfaction, there were still significant differences in the pain and motion of the affected shoulder when compared with those of the unaffected, contralateral shoulder (p < 0.00001). At the final outcome evaluation, the DASH scores demonstrated limitations when compared with known population norms, whereas the profiles of the SF-36 were comparable with those of age and gender-matched control populations. Prior treatment with physical therapy and a Workers' Compensation claim or pending litigation were the only variables that were associated with the eventual need for manipulation or capsular release. Male gender and diabetes mellitus were associated with worse motion at the final evaluation. Patients with a greater severity of pain with activity at the initial evaluation had significantly lower DASH scores at the final evaluation, and patients with lower initial scores on the Simple Shoulder Test had comparatively lower scores on the Simple Shoulder Test at the outcome evaluation. CONCLUSIONS: The vast majority of patients who have phase-II idiopathic adhesive capsulitis can be successfully treated with a specific four-direction shoulder-stretching exercise program. Although measurable limitations and deficiencies were noted at the outcome evaluation, these appeared to be acceptable to most of the patients and did not affect their general health status. Patients with more severe pain and functional limitations before treatment had relatively worse outcomes. More aggressive treatment such as manipulation or capsular release was rarely necessary, and the efficacy of early use of these treatments should be further studied. PMID- 11057468 TI - Osteonecrosis of the femoral head treated with cementless total hip arthroplasty. AB - BACKGROUND: The treatment of young patients who have osteonecrosis of the femoral head associated with collapse or substantial secondary degeneration remains a therapeutic challenge, with total hip arthroplasty being a treatment of choice. However, concerns about the durability of the results of hip arthroplasty in this population necessitate long-term evaluation of this treatment option. To determine its advantages and limitations, we evaluated the results of cementless total hip arthroplasty in a consecutive series of young patients with advanced osteonecrosis. METHODS: We reviewed the results of fifty-five consecutive primary total hip arthroplasties, after an average of 117 months of follow-up, in forty five patients with a preoperative diagnosis of advanced osteonecrosis of the femoral head (Ficat and Arlet stage III or IV). The average age was thirty-one years (range, twenty-one to forty years) at the time of the operation. We collected data prospectively with the use of patient questionnaires and radiographs. RESULTS: Five patients died and one patient was lost to follow-up before the time of the minimum five-year follow-up; this left forty-eight hips in thirty-nine patients for inclusion in the study. Ten (21 percent) of the forty eight hips required revision. No revisions were due to aseptic failure of the femoral component. Of the remaining twenty-nine patients (thirty-eight hips), twenty-seven (93 percent) reported few or no functional limitations and twenty three (79 percent) could walk an unlimited distance at the time of the latest follow-up. Pain was absent or mild in twenty-five patients (86 percent). Twenty three patients (79 percent) were employed full-time. Radiographically, thirty seven femoral components (97 percent) were bone-ingrown and the remaining component was judged to be fibrous stable. All thirty-eight acetabular components were bone-ingrown. CONCLUSIONS: Cementless total hip arthroplasty remains a reasonable treatment option for advanced osteonecrosis of the femoral head. Wear of the bearing surface continues to limit the long-term success rate, but we are encouraged by the predictable long-term stability of the bone-implant interface achieved with cementless fixation. These results compare favorably with those of published reports of total hip arthroplasty with cement in younger patients with osteonecrosis. PMID- 11057469 TI - The accuracy and reproducibility of radiographic assessment of stress-shielding. A postmortem analysis. AB - BACKGROUND: Although periprosthetic bone loss remains a major concern in total hip arthroplasty, radiographic assessment of such loss is both difficult and subjective. In the present study, we assessed the ability of orthopaedic surgeons to reproducibly recognize changes in periprosthetic bone density on radiographs. We hypothesized that assessment of periprosthetic bone loss on plain radiographs is not reliable enough to justify its use in outcomes research. METHODS: Twenty nine unilateral total hip replacements and the surrounding bone were retrieved at autopsy, and radiographs were made; radiographs of the contralateral, normal femur were also made after implantation of an identical prosthesis and used as a control. Three orthopaedic surgeons independently examined the specimen radiographs and classified bone loss in each of sixteen femoral zones. Bone loss was recorded as present if the bone of the femur that had had in vivo implantation showed evidence of cortical thinning, increased porosity, or decreased density (either cortical or trabecular) when compared with the control femur. The kappa coefficient was used to quantify interobserver and intraobserver reproducibility in determining bone loss for the 464 zones examined and in determining the Engh and Bobyn stress-shielding classification of each femur. In fourteen femoral pairs, bone loss was also quantified with dual-energy x-ray absorptiometry, and the resulting value was then compared with the bone-loss classification that had been determined radiographically. RESULTS: First, the surgeons agreed on the presence or absence of bone loss in 73 percent (337) of the 464 zones. The interobserver kappa value of 0.58 denoted only good reproducibility. The intraobserver reproducibility was better; the surgeon's initial evaluation of bone loss agreed with his second evaluation for 90 percent of the zones (kappa = 0.74). Second, the three surgeons agreed on the degree of stress-shielding, according to the Engh and Bobyn classification, in 66 percent (nineteen) of the twenty-nine femora. The kappa value for this comparison was only 0.27, indicating marginal reproducibility. Third, although there was some agreement among reviewers when there was 20 to 60 percent reduction in bone mineral content as determined with dual-energy x-ray absorptiometry, excellent agreement among the examiners (kappa = 0.85) was not achieved until bone loss averaged 70 percent. CONCLUSIONS: On the basis of these results, we suggest caution in interpreting results from studies of femoral bone loss that have used plain radiographic analysis if the authors have not provided interobserver reliability data. We question the utility of evaluating periprosthetic bone loss on radiographs, since the loss is not reproducibly recognized until 70 percent of the bone is gone. PMID- 11057470 TI - Scintigraphic assessment of the rotated femoral head after transtrochanteric rotational osteotomy for osteonecrosis. AB - BACKGROUND: The purpose of this study was to assess the usefulness of bone scintigraphy in predicting progressive collapse of the femoral head after transtrochanteric rotational osteotomy for the treatment of osteonecrosis of the femoral head. METHODS: We studied thirty-three hips in thirty patients with osteonecrosis of the femoral head who had undergone transtrochanteric rotational osteotomy. There were twenty male and ten female patients, with a mean age of 34.4 years at the time of the operation. The mean duration of follow-up was 10.0 years. According to the staging system of Ficat and Arlet, there were nineteen stage-2 hips and fourteen stage-3 hips at the time of the operation. Conventional anteroposterior and lateral radiographs were assessed. In addition, bone scans were performed at three weeks after the operation to predict the outcome with regard to the rotated femoral head. On the basis of the location of low scan activity within the femoral head, the scintigraphic findings were classified into one of two categories: type A if there was no low scan activity in the weight bearing area of the femoral head or type B if low scan activity occupied the entire weight-bearing area. Six hips with collapse were studied histologically. RESULTS: Postoperative scintiscans revealed sixteen type-A hips and seventeen type-B hips. Of the type-A hips, only three exhibited progressive collapse of the femoral head after the osteotomy, whereas fourteen of the type-B hips exhibited progressive collapse. A significant association was found between the postoperative scintigraphic findings and the final radiographic result (p < 0.01). CONCLUSIONS: Bone scintiscans made three weeks after transtrochanteric rotational osteotomy were useful for predicting the final clinical result. PMID- 11057471 TI - Evaluation of periprosthetic bone-remodeling after cementless total hip arthroplasty. The influence of the extent of porous coating. AB - BACKGROUND: Total hip arthroplasty changes the levels of stress within the proximal part of the femur, and the femur remodels adjacent to the prosthesis. The stem size and the initial bone-mineral density around the distal portion of the stem affect postoperative bone-remodeling after the insertion of a fully porous-coated metal-cancellous prosthesis. The purpose of this study was to evaluate the influence of the extent of porous coating of this prosthesis on femoral bone-remodeling. METHODS: A longitudinal examination of sixty-one hips in fifty-four patients was performed. Thirty-one hips in twenty-seven patients with a fully porous-coated stem (Group A) and thirty hips in twenty-seven patients with a proximally porous-coated stem (Group B) were followed for twenty-four to thirty months. Periprosthetic bone-mineral density was measured with dual-energy x-ray absorptiometry at specific intervals after the operation. RESULTS: In both groups, the greatest loss of bone-mineral density, compared with the initial (three-week) value, was approximately 20 percent in zone 7 at twelve to eighteen months. In other zones, bone-remodeling appeared to cease by twelve months. At the last follow-up evaluation, the loss of bone-mineral density in the distal and middle regions in Group A was significantly greater than that in Group B (p < 0.01 for zone 3 and p < 0.05 for zone 6). In contrast, with the numbers available, there were no significant differences in loss of bone-mineral density in the proximal regions (zones 1 and 7) between the two groups at any follow-up period. CONCLUSIONS: The extent of porous coating affects bone-remodeling in the distal periprosthetic region rather than in the proximal region. The results in the present report are specific to the particular implants that were studied. PMID- 11057472 TI - Multiplier method for predicting limb-length discrepancy. AB - BACKGROUND: In patients with a congenital or developmental limb-length discrepancy, the short limb grows at a rate proportional to that of the normal, long limb. This is the basis of predicting limb-length discrepancy with existing methods, which are complicated and require multiple data points. The purpose of our study was to derive a simple arithmetic formula that can easily and accurately predict limb-length discrepancy at skeletal maturity. METHODS: Using available databases, we divided the femoral and tibial lengths at skeletal maturity by the femoral and tibial lengths at each age for each percentile group. The resultant number was called the multiplier. Using the multiplier, we derived formulae to predict the limb-length discrepancy and the amount of growth remaining. We verified the accuracy of these formulae by evaluating two groups of patients with congenital shortening who were managed with epiphysiodesis or limb lengthening. We also calculated and compared the multipliers for other databases according to radiographic, clinical, and anthropological lower-limb measurements. RESULTS: The multipliers for the femur and tibia were equivalent in all percentile groups, varying only by age and gender. Because congenital limb-length discrepancy increases at a rate proportional to growth, the discrepancy at maturity can be calculated as the current discrepancy times the multiplier for the current age and the gender. This calculation can be performed with use of a single measurement of limb-length discrepancy. For progressive developmental (noncongenital) discrepancies, the discrepancy at skeletal maturity can be calculated as the current discrepancy plus the growth inhibition times the amount of growth remaining. The timing of the epiphysiodesis can also be calculated with the multiplier. The predictions made with use of the multiplier method correlated well with those made with use of the Moseley method as well as with the actual limb-length discrepancy in both the limb-lengthening and epiphysiodesis groups. The multipliers derived from the radiographic, clinical, and anthropological measurements of femora and tibiae were all similar to each other despite differences in race, ethnicity, and generation. CONCLUSIONS: The multiplier method allows for a quick calculation of the predicted limb-length discrepancy at skeletal maturity, without the need to plot graphs, and is based on as few as one or two measurements. This method is independent of percentile groups and is the same for the prediction of femoral, tibial, and total-limb lengths. The multiplier values are also independent of generation, height, socioeconomic class, ethnicity, and race. We verified the accuracy of this method clinically by evaluating patients who had been managed with limb-lengthening or epiphysiodesis. The method was also comparable with or more accurate than the Moseley method of limb-length prediction. PMID- 11057473 TI - Flexible intramedullary nailing for the treatment of unicameral bone cysts in long bones. AB - BACKGROUND: Unicameral bone cyst is characterized by its tenacity and risk of recurrence. Pathological fracture is common and is often the presenting symptom. The objective of the present study was to evaluate the results of flexible intramedullary nailing for the treatment of a unicameral bone cyst with or without a pathological fracture. METHODS: Flexible intramedullary nailing for the treatment of a unicameral bone cyst was performed in thirty-two patients. Thirty of these patients presented with a pathological fracture; twenty-four were managed immediately with intramedullary nailing, and the other six had been managed conservatively at other clinics before they were referred to our department. The remaining two cysts were detected incidentally. The cyst was located in the humerus in twenty-one patients, in the femur in nine, and in the radius in two. The mean age of the patients at the time of surgery was 9.8 years, and the mean duration of follow-up was 53.7 months. Radiographic evaluation was performed according to the criteria of Capanna et al., and the cyst was classified as completely healed, healed with residual radiolucency (osteolysis), recurred, or having no response. RESULTS: The healing period ranged from three to 105 months. Fourteen cysts healed completely, and sixteen healed with residual radiolucent areas visible on radiographs. There was recurrence of two cysts that had healed with residual radiolucency. All of the cysts in the present study responded to treatment. A change of nails was necessary in nine patients, as the nails had become too short after bone growth. No major complications were observed. CONCLUSIONS: Flexible intramedullary nailing provides early stability, which allows early mobilization and thus obviates the need for a plaster cast and decreases the prevalence of the most common complication: a pathological fracture. This method of treatment also allows for an early return to normal activity. PMID- 11057474 TI - Autodigestion of a hamstring anterior cruciate ligament autograft following thermal shrinkage. A case report and sentinel of concern. PMID- 11057475 TI - The use of pedicle-screw internal fixation for the operative treatment of spinal disorders. AB - Pedicle screws have dramatically improved the outcomes of spinal reconstruction requiring spinal fusion. Short-segment surgical treatments based on the use of pedicle screws for the treatment of neoplastic, developmental, congenital, traumatic, and degenerative conditions have been proved to be practical, safe, and effective. The Funnel Technique provides a straightforward, direct, and inexpensive way to very safely apply pedicle screws in the cervical, thoracic, or lumbar spine. Carefully applied pedicle-screw fixation does not produce severe or frequent complications. Pedicle-screw fixation can be effectively and safely used wherever a vertebral pedicle can accommodate a pedicle screw--that is, in the cervical, thoracic, or lumbar spine. Training in pedicle-screw application should be standard in orthopaedic training programs since pedicle-screw fixation represents the so-called gold standard of spinal internal fixation. PMID- 11057476 TI - Workers' compensation: avoiding work-related disability. PMID- 11057477 TI - The emerging impact of the information age on orthopaedic surgery. Introduction. PMID- 11057478 TI - Development of a virtual reality arthroscopic knee simulator. PMID- 11057479 TI - Electronic resources for the orthopaedic surgeon. PMID- 11057480 TI - Implementation of a computer-based patient record and an outcomes data-collection system at the Department of Orthopaedic Surgery, University of Iowa. PMID- 11057481 TI - The value and promise of patient databases in orthopaedic surgery. PMID- 11057483 TI - Ethics in practice: residency training. PMID- 11057482 TI - Lengthening of the achilles tendon in diabetic patients. PMID- 11057484 TI - Ethics in practice: residency training. PMID- 11057485 TI - Efficacy of the patellar tendon-bearing cast. PMID- 11057486 TI - Misrepresentation of research criteria by orthopaedic residency applicants. PMID- 11057487 TI - Collarless, polished, tapered stem failure. PMID- 11057488 TI - Collarless, polished, tapered stem failure. PMID- 11057489 TI - Adverse events associated with autologous and allogenic blood transfusion. PMID- 11057490 TI - Pedicle-screw placement. PMID- 11057491 TI - Proximal radioulnar synostosis. PMID- 11057492 TI - Pregnancy-related avascular necrosis. PMID- 11057493 TI - Stulberg classification system. PMID- 11057494 TI - Somatosensory evoked potentials. PMID- 11057495 TI - Osteolysis and ceramic bearing surfaces. PMID- 11057496 TI - Osteolysis and ceramic bearing surfaces. PMID- 11057497 TI - Osteolysis and ceramic bearing surfaces. PMID- 11057498 TI - Clinical trial evaluating a new hub device designed to prevent catheter-related sepsis. AB - A new commercial hub device designed to minimise catheter-related infections was evaluated in a prospective, randomised trial in the intensive care and surgical units of the Hospital de Tortosa Verge de la Cinta in patients in whom the central venous catheters were expected to remain indwelling for at least 7 days. The assessments conducted at catheter withdrawal included cultures of the skin at the catheter site and cultures of the catheter tip and the catheter hubs; moreover, in cases of suspected catheter-related sepsis, samples of peripheral blood and infusion solutions were also cultured. Of the 130 catheters evaluated, 26 (20%) were withdrawn because of suspected catheter-related sepsis; 10 (15%) were in the control group and 16 (24%) in the new product group. Catheter-related sepsis was diagnosed in nine patients, six of whom were in the new product group and three in the control group; all infections in the former group and only one in the latter group were caused by the catheter connection. The rates of catheter hub colonisation (10 cfu) and catheter colonisation (15 cfu in semiquantitative culture and/or >1,000 cfu in quantitative culture) of hub origin were not significantly different between the groups (15 cases in the control group vs. 20 cases in the new product group, and 5 cases in the control group vs. 11 cases in the new product group, respectively). The data indicate that the use of the new catheter hub device is no more effective in preventing catheter-related infection than standard good clinical procedures. PMID- 11057499 TI - Epidemiology of yeast colonization in the intensive care unit. AB - In order to investigate the epidemiology of colonization and possible transmission of yeasts among patients and healthcare workers in adult intensive care units (ICUs), 194 patients were followed for a mean of 9 +/- 11 days and 63 healthcare workers were followed for a mean of 132 +/- 52 days. Among the patients, 142 (73%) were colonized by yeast, with Candida albicans being the species most commonly recovered. Most patients (65%) were already colonized with yeast upon admission to the intensive care unit; only 17% became colonized after admission. Persistent colonization occurred in 51 (55%) of 92 patients who had more than three cultures performed; in 75% of them, colonization persisted with the same strain of Candida albicans or Candida glabrata. Bacterial infection in the month preceding entry into the ICU was the only risk factor significantly associated with yeast colonization. Among the healthcare workers, yeasts were isolated from 42 (67%). Candida albicans was most frequently recovered from the oropharynx (19% of occasions), and Candida parapsilosis was most frequently found on hands (8% of occasions). Persistent colonization of the oropharynx occurred in only six healthcare workers, and none had persistence of yeasts on hands. In this non-outbreak setting, 5 (4%) of 123 patient/healthcare worker interactions that were linked epidemiologically yielded the same strain of Candida albicans, providing evidence for possible cross-transmission. No similar link was found between healthcare worker-patient interactions and colonization with Candida glabrata or Candida parapsilosis. PMID- 11057500 TI - Transmission of Pneumocystis carinii disease from immunocompetent contacts of infected hosts to susceptible hosts. AB - Pneumocystis carinii organisms constitute a large group of heterogeneous atypical microscopic fungi that are able to infect immunocompromised mammals by an airborne route and to proliferate in their lungs, inducing Pneumocystis carinii pneumonia. This pneumonia remains a crucial epidemiological challenge, since neither the source of Pneumocystis carinii infection in humans nor the process by which humans become infected has been clearly established. Polymerase chain reaction (PCR) assays have shown that profoundly immunosuppressed patients without pneumocystosis can be subclinically infected with Pneumocystis. Other PCR based studies have suggested that healthy immunocompetent hosts are not latent carriers of the parasite. However, recent reports have indicated that Pneumocystis carinii can persist for limited periods in the lungs of convalescent rats after recovery from corticosteroid-induced pneumocystosis, and also that immunocompetent mammals can be transiently parasitized by Pneumocystis carinii after close contact with hosts with Pneumocystis carinii pneumonia. Can transiently parasitized hosts be a source of infection for immunosuppressed hosts? In order to investigate this important clinical question, the ability of immunocompetent BALB/c mice, which were carrying subclinical levels of Pneumocystis carinii, to transmit the infection by the airborne route to highly susceptible, uninfected mice with severe combined immunodeficiency was studied. The results indicated that the immunocompetent mice, transiently parasitized by Pneumocystis carinii organisms after close contact with Pneumocystis carinii infected mice, were able to transmit the infection to Pneumocystis carinii-free mice with severe combined immunodeficiency. PMID- 11057501 TI - Enteric fever and other extraintestinal salmonellosis in University Hospital, Nottingham, UK, between 1980 and 1997. AB - The clinical spectrum of extraintestinal salmonellosis comprises enteric fever (typhoid and paratyphoid) and invasive infections due to nontyphoidal salmonellae. This study describes the clinical spectrum, management and outcome of all confirmed cases of extraintestinal salmonellosis in patients admitted to University Hospital, Nottingham, UK, between 1980 and 1997. There were 142 cases (children, 42; adults, 100) of extraintestinal salmonellosis, of which 38 (children, 20; adults, 18) were enteric fever, consisting of 21 cases of typhoid, 12 of paratyphoid A and five of paratyphoid B. All patients with typhoid and paratyphoid A fever were from Indian or Pakistani families and, except for two adults, all were considered to be previously fit. The outcome in patients with enteric fever was excellent, and there were no complications. Of the 104 patients (children, 22; adults, 82) with nontyphoidal salmonellosis, 69 were bacteraemic secondary to gastroenteritis, 10 were bacteraemic without an obvious focus of infection and 25 had focal infections. The three major sites of focal infections were meningitis in five infants, osteomyelitis in two children and three adults, and arterial infections in ten adults. The three most frequently isolated organisms were Salmonella enteritidis (40%), Salmonella typhimurium (25%) and Salmonella virchow (14%). Sixty-seven percent of these patients had underlying disease(s)/risk factors. In contrast to the outcome of enteric fever, there were 19 deaths (children, 2; adults, 17) in patients with nontyphoidal salmonellosis. Sixteen of the 17 adults who died were over the age of 60 years. Eight (25%) of 32 males over the age of 60 years with nontyphoidal Salmonella bacteraemia had arterial infections. In some patients, the diagnosis of Salmonella arterial infection is likely to be delayed or missed altogether if blood cultures are not obtained. Mortality in patients over the age of 60 years with nontyphoidal Salmonella infections was 28%. PMID- 11057502 TI - Role of antiretroviral therapy in long-term survival of patients with AIDS related pulmonary aspergillosis. AB - The aim of the study presented here was to describe the clinical presentation and outcome of invasive pulmonary aspergillosis (IPA) in a cohort of HIV-infected patients and to assess factors associated with survival by means of a longitudinal study of all HIV-infected adults with IPA who attended the infectious diseases service of a tertiary center between January 1985 and December 1998. The outcome measure was the time to death after diagnosis of IPA. The endpoint for data collection and survival analysis was 31 December 1998. Nineteen of 1,605 HIV-infected patients were identified, resulting in an overall IPA attack rate of 1.12%. Most patients had AIDS (95%). Unilateral cavitary disease was the most frequent radiographic presentation (37%). Median survival was 148 days (95% confidence interval [95% CI], 0-402), but it exceeded 12 months in 37%. In patients in whom antiretroviral treatment (ART) was initiated or modified in relation to the IPA diagnosis, median survival increased to 906 days (95% CI, 754-1,058; 1-year survival, 83%) compared with 86 days in those who did not take any ART (95% CI, 55-117; 1-year survival, 8%; P=0.0002). Survival was even longer when ART changes included only nucleoside reverse transcriptase inhibitors, prior to the availability of protease inhibitors (833 days, 95% CI, 369-1,297; 1-year survival 75%; P=0.003). Three (16%) patients are still alive after a mean time of 36 months postdiagnosis. Although IPA is a life-threatening complication of advanced AIDS, it does not always lead to short-term death. An adequate management of HIV infection together with antifungal treatment may prolong survival and, as described for other AIDS-related disorders, a significant decrease in its occurrence can be expected. PMID- 11057503 TI - Clinical evaluation of a novel chromogenic agar dipslide for diagnosis of urinary tract infections. AB - The performance of a novel dipslide (DipStreak; Novamed, Israel) consisting of chromogenic agar (Uriselect 3; Sanofi Pasteur, France) and blood agar media was evaluated prospectively and compared to that of conventional urine culture for the diagnosis of urinary tract infection. A total of 1070 clean-catch urine specimens obtained from 251 hospitalized patients and 819 outpatients were processed. The overall performance of the DipStreak was as follows: sensitivity, 95.7%; specificity, 99.2%; agreement, 89.8%; accuracy, 98%; positive predictive value, 98.5%; and negative predictive value, 97.7%. A total of 270 urine specimens were positive by both DipStreak and conventional culture. The chromogenic agar allowed rapid identification of organisms in 211 (78.1%) cultures, while isolates in the other 59 (21.9%) cultures remained unidentified. The results indicate that the DipStreak device coupled with the Uriselect 3 agar represents a convenient and accurate method for inoculation of urine specimens, quantitation of bacteria, diagnosis of significant bacteriuria, and presumptive identification of isolates. PMID- 11057504 TI - Infrequency of pulmonary microbial colonisation prior to respiratory disease in HIV-infected individuals. AB - To determine whether organisms are present in the HIV-infected lung prior to clinical respiratory disease, a cross-sectional bronchoscopic comparative analysis of 39 asymptomatic HIV-positive subjects and 31 healthy controls with 2 year prospective bronchoscopic monitoring of the HIV study group was performed. Pathological examination of bronchoalveolar lavage (BAL) fluid using standard microbiological techniques was undertaken. Organisms were recovered from similar numbers of HIV-positive and control subjects (7 of 39 and 3 of 31) and comprised predominantly scanty growths of bacteria. Five subjects developed respiratory disease during follow-up. Repeat BAL was performed in 11 asymptomatic HIV positive patients; no relationship was found between the organisms isolated at the two procedures. The findings suggest that the asymptomatic HIV-positive lung is not a frequent site of either microbial colonisation or subclinical infection. This has implications for the understanding of the pathogenesis of HIV-related pulmonary disease. PMID- 11057505 TI - Surveillance of penicillin-resistant Streptococcus pneumoniae in one American metropolitan area, 1989-1998. AB - The emergence of invasive penicillin-resistant (PRSP) and multidrug-resistant (MDRP) Streptococcus pneumoniae was tracked from 1989 to 1998 in one medium-sized metropolitan area in the USA, encompassing western West Virginia, including Huntington, the only major city, and neighboring sections of Kentucky and Ohio. Capsular serotyping and antibiotic sensitivity tests were performed on 350 community-acquired isolates comprising 93.1% of all pneumococcal isolates identified. The incidence of PRSP increased from 3 to 10% during the 10 years of the study. Twenty-nine (22.1%) of 131 isolates of serotypes 6, 9, 14, 19, and 23 were PRSP (one-fourth were MDRP) and 1 (0.5%) of 219 other serotypes was PRSP (serotype 35). Invasive PRSP occurred most frequently in young children and in adults aged 80 years and older, 8.9 and 10.9 cases per 100,000 persons, respectively. PMID- 11057506 TI - Extraintestinal infection due to Hafnia alvei. AB - The aim of this study was to establish the clinical features of extraintestinal infections caused by Hafnia alvei. Over a 5-year period (1994-1998), data were collected regarding inpatients (n = 8) with nosocomial (n = 5) or community acquired (n = 3) infections caused by Hafnia alvei. The mean age of the patients was 47 +/- 21 years. Three patients had hospital-acquired urinary tract infections. Hafnia alvei also caused community-acquired cholangitis, cholecystitis, appendicitis, psoas abscess and prosthetic endocarditis. Hafnia alvei was susceptible to amoxicillin/clavulanic acid and to first-generation cephalosporins in two cases. Susceptibility to aminoglycosides, imipenem, cotrimoxazole, ciprofloxacin, piperacillin and cefotaxime was very good (8/8). Four patients required invasive treatment. PMID- 11057507 TI - Listeriosis in recipients of allogeneic bone marrow transplants from unrelated donors. AB - Two cases of listeriosis in patients submitted to matched unrelated donor bone marrow transplantation are reported. The patients developed listerial septicemia and listerial septicemia with meningitis and encephalitis 39 and 29 days after transplantation, respectively. Including the present two cases, 19 Listeria monocytogenes infections in related and unrelated donor allogeneic bone marrow transplant recipients have been reported to date. Infection occurred earlier in unrelated donor transplant recipients. Listeriosis is a rare complication in allogeneic bone marrow transplant recipients; however, the widespread practice of performing transplants from a donor-alternative to a human leukocyte antigen compatible sibling and, in this setting, the need for intensified immunosuppression may predict an increasing and earlier occurrence of listeriosis. PMID- 11057508 TI - Evaluation of the Bactec 960 automated nonradiometric system for isolation of mycobacteria from clinical specimens. AB - The Bactec MGIT 960 system (Becton Dickinson, USA), designed for the culture of mycobacteria, was compared with the Bactec 460 instrument and culture on two solid egg-based media using a total of 1024 clinical specimens. Mycobacteria could be identified from 99 (9.7%) specimens, 89 (90%) of which were identified by the Bactec 960 system, 90 (91%) by the Bactec 460 system, and 82 (83%) by culture on the two egg-based media. The Bactec 960 cultures became positive an average of 16.7 days after specimen collection, the Bactec 460 cultures 14.9 days after collection, and the cultures on egg-based media 26.2 days after collection. The Bactec 960 is a compact and highly automated nonradiometric system that may replace the Bactec 460 system. PMID- 11057509 TI - Assessment of optimal atmospheric conditions for growth of Helicobacter pylori. AB - The number and diameter of colonies of Helicobacter pylori isolates growing on agar plates were determined to compare five methods that produce a culture atmosphere. No catalyst was applied. No significant difference was found between two hydrogen-based kits that have a different capacity for production of H2. These hydrogen-based methods were significantly better than all others evaluated, including one kit that produces ascorbic acid that binds with oxygen. Growth was significantly improved when the atmosphere outside the plastic incubation jars was enriched with 10% CO2, but carbon dioxide enrichment alone (i.e., no reduction of the oxygen concentration) gave a very poor yield. The colony diameter was a sensitive and reliable measure of atmospheric conditions, as the mean intra- and interobserver difference between repeated readings was < or = 0.1 mm for 82% and < or = 0.2 mm for 95% of the isolates. PMID- 11057510 TI - High prevalence of GB virus C/hepatitis G virus RNA and antibodies in patients infected with human immunodeficiency virus type 1. AB - The prevalence of GB virus C (GBV-C)/ hepatitis G virus (HGV) RNA and antibodies to the structural E2 protein was investigated in a cohort of HIV-1 infected patients. Of 346 individuals, RNA was detected in 143 and E2 antibodies were detected in 73, for an overall prevalence of 62.4%. Intravenous drug use and homosexuality were identified as major transmission risk factors. GBV-C/HGV RNA prevalence was associated with hepatitis B coinfection, whereas antibodies to E2 were associated with older age and lower CD4+ cell counts. GBV-C/HGV infection was frequent in this group of HIV-infected patients and was associated with older age, lower CD4 + cell counts, and the presence of hepatitis B surface antigen. PMID- 11057511 TI - Two cases of pulmonary complications associated with a recently recognised Salmonella enteritidis phage type, 21b, affecting immunocompetent adults. PMID- 11057512 TI - Joint tuberculosis caused by Mycobacterium bovis in an immunocompetent patient. PMID- 11057513 TI - Distribution of hepatitis C virus genotypes in viremic patients with beta thalassemia. PMID- 11057514 TI - Molecular typing of Myroides odoratimimus (Flavobacterium odoratum) urinary tract infections in a Turkish hospital. PMID- 11057515 TI - Cocaine-seeking by rats: regulation, reinforcement and activation. AB - RATIONALE: In animal models of drug self-administration, response rates often decrease with dose suggesting that a regulative process may mask the reinforcing effects of the drug. OBJECTIVE: The purpose of the present experiments was to dissociate the role of regulative and reinforcement processes in intravenous cocaine self-administration by rats using a paradigm that explicitly distinguishes between drug-seeking and drug-taking. METHODS: Rats were trained to respond for intravenous cocaine (0.25 mg/infusion) under a heterogeneous chain (tandem FR1 RI 30 s) FR1 schedule of reinforcement using different levers in the first (seeking) and second (taking) links of the chain. After 10 days of training, rats were switched to one of three doses of cocaine (0.08, 0.25, or 0.5 mg/infusion) and self-administration patterns were recorded for a further ten sessions in experiment 1. In experiment 2, a time-out (TO) period (0, 4, or 12 min) was imposed between successive cycles of the chain schedule. Finally, the effect of allowing animals to perform a drug-taking response on subsequent drug seeking was assessed in experiment 3. RESULTS: Having verified that seeking responses for a conventional reinforcer (sucrose) were sensitive to changes in reward magnitude, experiment 1 demonstrated that the number of self-administered infusions was inversely related to dose whereas the latency to initiate drug seeking increased with dose. Variations in the cocaine dose had no reliable effect on the number of drug seeking response per cycle of the chain schedule. The effect of dose on the latency to initiate drug-seeking was reversed in experiment 2 with increasing TO periods. Moreover, at the longest TO period, drug seeking responses per cycle increased and the latency to initiate drug seeking decreased with dose. Experiment 3 showed that the latency to drug-seek for the low dose was reduced dramatically when the first drug-seeking response was preceded by a drug-taking response, even when this response did not produce a drug infusion. CONCLUSIONS: The overall pattern of results suggests that drug seeking and drug-taking are controlled by three interacting processes: a regulative process depresses drug-seeking in the short-term; behavioral activation enhances drug-seeking and is sustained over longer intervals by higher drug doses; the reinforcing effect of cocaine increases with dose once the satiety producing effects of the drug dissipate. PMID- 11057516 TI - Effects of sex and the estrous cycle on regulation of intravenously self administered cocaine in rats. AB - RATIONALE: Previous research with both humans and animals suggests that there are sex differences in cocaine self-administration; in rodents, ovarian hormones may underlie these differences. OBJECTIVES: A two-lever drug self-administration procedure was used to compare regulation of intravenously self-administered cocaine in male and female rats and among females in different phases of the estrous cycle. METHODS: Eleven female and seven male age-matched Wistar rats were trained to self-administer nine doses of cocaine (0.0-2.4 mg/kg) during daily 5-h sessions. Experimental test chambers were equipped with two levers and associated stimulus lights. A response on the lever with stimuli signaling an increase in cocaine dose increased the infusion duration by 3 s, and a response on the other lever decreased the infusion duration by 3 s. RESULTS: After responding for cocaine stabilized, regulation was disrupted more in females than in males (r2=78.9, r2=92.6, respectively) with the greatest disruption observed in females during the estrus phase (r2=48.5). Mean dose size varied considerably for males and for females in the metestrus/diestrus and proestrus phases; however, estrus females responded almost exclusively on the lever associated with an increase in cocaine dose. CONCLUSIONS: These findings indicate sex differences in the regulation of cocaine self-administration, and they suggest that ovarian hormones may be responsible for the observed sex differences. PMID- 11057517 TI - Psychological stress, drug-related cues and cocaine craving. AB - RATIONALE: While several environmental situations may produce cocaine craving, there is little research on whether patterns of drug cue reactivity are similar across different environmental situations. OBJECTIVE: This study examined whether two different environmental situations, psychological stress and drug cues, produce similar or varying patterns of cue reactivity in 20 cocaine dependent individuals. METHODS: All subjects participated in a single laboratory session and were exposed to stress, drug cues and neutral-relaxing imagery conditions. Cocaine and alcohol craving, emotion state ratings, subjective anxiety, heart rate and salivary cortisol measures were assessed. RESULTS: Significant increases in cocaine and alcohol craving were observed with stress and drug cues imagery but not with neutral-relaxing imagery. In addition, stress and drug cues situations produced similar increases in subjective anxiety, heart rate and salivary cortisol levels. Significant increases in negative emotion ratings and decreases in positive emotion ratings were found for stress and drug cues conditions as compared to the neutral condition. CONCLUSIONS: The findings indicate that a similar and comparable pattern of cue reactivity is induced by stress and drug cue manipulations. Furthermore, the comparable increases in subjective anxiety and negative affect observed with stress-induced and drug cue induced craving provides support for the negative reinforcement model of drug craving and relapse. The negative affectivity co-occurring with the craving state appears to be an important target in the development of new treatments for cocaine dependence. PMID- 11057518 TI - Fenfluramine effects on impulsivity in a sample of adults with and without history of conduct disorder. AB - RATIONALE: The role of serotonin in impulsivity was examined by administering the serotonin-releasing drug, d,l-fenfluramine, and measuring effects on impulsive responding of male subjects with and without a history of conduct disorder (CD) under controlled laboratory conditions. METHODS: Five adult male subjects with a history of CD and five matched controls were recruited into a study to determine the acute effects of d,l-fenfluramine on a laboratory measure of impulsive behavior. This laboratory measure, based upon delay of gratification, presented subjects with choices between a small reward after a short delay and a larger reward after a longer delay. Impulsive behavior was indicated by frequent choices for the smaller reward. RESULTS: Acute oral doses of d,l-fenfluramine (0.21, 0.42, and 0.85 mg/kg) produced decreases in the number of impulsive choices in all subjects with a history of CD, but had no effect on the control subjects. CONCLUSIONS: This data suggests that a deficit in serotonin and/or dopamine may play a role in impulsivity in CD subjects, and drugs which act to reduce this biological deficit can reduce impulsiveness. PMID- 11057519 TI - Different effects of 5-HT1A receptor agonists and benzodiazepine anxiolytics on the emotional state of naive and stressed mice: a study using the hole-board test. AB - OBJECTIVES: The effects of 5-HT(1A) receptor agonists on the emotional behavior of naive or stressed mice were examined and compared with those of benzodiazepine anxiolytics. METHODS: Changes in the emotional state of mice were evaluated in terms of changes in exploratory activity, i.e. total locomotor activity, numbers and duration of rearing and head-dipping and latency to the first head-dipping, using an automatic holeboard apparatus. RESULTS: The 5-HT(1A) receptor full agonists flesinoxan (0.03-1 mg/kg, IP) and 8-OH-DPAT (0.03-1 mg/kg, IP), and the partial agonist buspirone (0.3-10 mg/kg, IP) dose-dependently decreased all of the exploratory behaviors. Significant decreases in both the number and duration of head-dips, and an increase in the latency to head-dipping were observed at 30 min after exposure to acute restraint stress (60 min). These emotional changes were scarcely improved by post-stress treatment with 5-HT(1A) receptor agonists, at doses that alone did not produce a significant behavioral effect. In contrast, pretreatment with flesinoxan (0.1-1 mg/kg, IP) or 8-OH-DPAT (0.1-1 mg/kg, IP) 24 h prior to exposure to stress dose-dependently suppressed the decrease in various exploratory behaviors that was observed immediately after the exposure to acute restraint stress. Moreover, pretreatment with buspirone (1-10 mg/kg, IP) 24 h prior to exposure to stress also significantly suppressed the decrease in rearing behavior and the increase in head-dip latency. However, changes in the emotional response to stress stimuli were not observed in mice that had been pretreated with the benzodiazepine anxiolytics diazepam (0.1-1 mg/kg, IP) and chlordiazepoxide (2-8 mg/kg, IP). CONCLUSIONS: The present study clearly demonstrates that the behavioral effects of 5-HT(1A) receptor agonists in both naive and stressed mice were quite different from those of benzodiazepine anxiolytics, as previously reported by us. Notably, 5-HT(1A) receptor agonists but not benzodiazepine anxiolytics protect against various emotional changes produced by stress stimuli, and the results suggest that activation of 5-HT(1A) receptors may facilitate some mechanism(s) involved in the recognition of and/or ability to cope with stressful situation. PMID- 11057520 TI - Effects of low doses of caffeine on cognitive performance, mood and thirst in low and higher caffeine consumers. AB - RATIONALE: Caffeine is present in many widely consumed drinks and some foods. In the fairly extensive literature on the psychostimulant effects of caffeine, there are few dose-response studies and even fewer studies of the effects of doses of caffeine lower than 50 mg (the range of the amounts of caffeine contained in, for example, a typical serving of tea or cola). OBJECTIVE: This study measured the effects of 0, 12.5, 25, 50 and 100 mg caffeine on cognitive performance, mood and thirst in adults with low and moderate to high habitual caffeine intakes. METHODS: This was a double-blind, within-subjects study. Following overnight caffeine abstinence, participants (n=23) completed a test battery once before and three times after placebo or caffeine administration. The test battery consisted of two performance tests, a long duration simple reaction time task and a rapid visual information processing task, and a mood questionnaire (including also an item on thirst). RESULTS: Effects on performance and mood confirmed a psychostimulant action of caffeine. All doses of caffeine significantly affected cognitive performance, and the dose-response relationships for these effects were rather flat. The effects on performance were more marked in individuals with a higher level of habitual caffeine intake, whereas caffeine increased thirst only in low caffeine consumers. CONCLUSIONS: After overnight caffeine abstinence, caffeine can significantly affect cognitive performance, mood and thirst at doses within and even lower than the range of amounts of caffeine contained in a single serving of popular caffeine-containing drinks. Regular caffeine consumers appear to show substantial tolerance to the thirst-increasing but not to the performance and mood effects of caffeine. PMID- 11057521 TI - Increased dopamine D2 receptor binding after long-term treatment with antipsychotics in humans: a clinical PET study. AB - RATIONALE: Dopamine D2 receptor upregulation in the striatum is regularly seen in response to the administration of traditional antipsychotics in animal experiments. This is associated with hyperactivity and, for this reason, D2 receptor upregulation has long been postulated as central to tardive dyskinesia (TD). OBJECTIVE: Using positron emission tomography (PET), the present study attempted to determine whether antipsychotic-induced D2 receptor up-regulation also occurs in humans. METHODS: The long-term effects of traditional and novel antipsychotics on dopamine D2 receptors were investigated in nine subjects meeting DSM-IV criteria for schizophrenia who were deemed eligible for temporary treatment washout. Subjects had been treated with traditional antipsychotics (haloperidol n=3, perphenazine n=1) and novel antipsychotics (risperidone n=3, olanzapine n=2) in the moderate to high dosage range. Fourteen days after treatment withdrawal, the binding potentials (BPs) of dopamine D2 receptors were measured using 11[C] raclopride. The obtained BPs were compared to the BPs from antipsychotic-naive control subjects with schizophrenia. RESULTS: There was a significant increase in the D2 BP in both groups combined that reached 34%. The increases in the D2 BPs in the groups treated with conventional and novel antipsychotics were 37% and 31%, respectively. Significantly, the patients showing the highest degree of D2 receptor upregulation (98%) developed severe and persistent TD shortly after being started on a new antipsychotic with low affinity for D2 receptors. CONCLUSION: This study demonstrates for the first time, using in vivo neuroreceptor imaging, that dopamine D2 receptor binding is increased after long-term treatment with antipsychotics in humans. The data suggest that both traditional and novel antipsychotics with high affinity for dopamine D2 receptors are associated with a substantial increase in D2 receptor binding. The present data in humans agree well with animal data that implicate D2 receptor-mediated mechanisms in motor hyperactivity. PMID- 11057522 TI - Characterization of the discriminative stimulus effects of GABA(A) receptor ligands in Macaca fascicularis monkeys under different ethanol training conditions. AB - RATIONALE: The current study was designed to extend our knowledge of the GABA(A) receptor system in mediating discriminative stimulus effects of ethanol in non human primates. OBJECTIVES: To characterize the discriminative stimulus effects of ethanol, pentobarbital, midazolam, muscimol and morphine in male and female monkeys under different ethanol training conditions. METHODS: Adult male (n=8) and female (n=10) Macaca fascicularis monkeys were divided into four groups and trained to discriminate 1.0 g/kg ethanol (n=8) versus water or 2.0 g/kg ethanol (n=10) versus water in a 2x2 design with training dose and sex as main group factors. Solutions were administered intragastrically (20% ethanol w/v) and responding was maintained under a fixed-ratio schedule of food reinforcement. Dose-response determinations of ethanol, pentobarbital, midazolam, muscimol and morphine were made under the training condition of 30 min pretreatment interval. The ethanol pretreatment interval in training sessions was then increased to 60 min and the effects of ethanol, pentobarbital and midazolam were redetermined. RESULTS: Training dose influenced the ED50 of ethanol to produce substitution under both pretreatment intervals and pentobarbital to produce substitution under the 30-min pretreatment training interval. There were no group differences in sensitivity to midazolam. The potency of the ligands to produce ethanol substitution was consistent across groups with midazolam>pentobarbital>ethanol. There were no sex differences in substitution of the ligands for ethanol. Blood ethanol concentrations at the onset of ethanol training sessions were higher in the 2.0 g/kg groups and under longer pretreatment times, but were not different on the basis of sex. CONCLUSIONS: Pentobarbital and midazolam produce ethanol like discriminative stimulus effects in male and female cynomolgus monkeys suggesting a significant GABA(A) component mediating the behavioral effects of ethanol. There was limited evidence that training dose of ethanol influenced substitution pattern of the GABA(A) ligands in cynomolgus monkeys, unlike previous findings in rats. Finally, there appear to be no sex differences in the profile of GABA(A) mechanisms involved in the discriminative stimulus effects of ethanol. PMID- 11057523 TI - Effect of intra-accumbens dopamine receptor agents on reactivity to spatial and non-spatial changes in mice. AB - RATIONALE: Some evidence suggests an involvement of nucleus accumbens in spatial learning. However, it is controversial whether the mesoaccumbens dopaminergic pathways play a specific role in the acquisition of spatial information. OBJECTIVE: The goal of these experiments was to investigate the effect of dopaminergic manipulations in the nucleus accumbens on a non-associative task designed to estimate the ability to encode/transmit spatial and non-spatial information. METHODS: The effects of focal administrations of the D1 and D2 dopamine receptor antagonists, SCH 23390 (6.25, 12.5, 50 ng/side) and sulpiride (12.5, 50, 100 ng/side), respectively, and dopamine (DA; 1.25 and 2.5 microg/side) into the nucleus accumbens were studied on reactivity to spatial and non-spatial changes in an open field with objects. RESULTS: Both SCH 23390 and sulpiride impaired reactivity to spatial change. However, several differences were found in the effects induced by the two DA antagonists. SCH 23390 did not affect locomotor activity and only slightly impaired exploration of the novel object. On the contrary, the D2 antagonist, induced a general, dose-dependent, impairment on all variables measured. Local administration of DA increased locomotor activity, but did not affect reactivity to spatial and non-spatial changes. CONCLUSIONS: These results demonstrate a facilitatory role of mesoaccumbens dopamine in the acquisition of spatial information. Moreover, they suggest that nucleus accumbens D1 DA receptors, play a more selective role in the modulation of spatial learning than accumbens D2 DA receptors. PMID- 11057524 TI - Chronic food restriction in rats augments the central rewarding effect of cocaine and the delta1 opioid agonist, DPDPE, but not the delta2 agonist, deltorphin-II. AB - RATIONALE: Chronic food restriction augments the self-administration and locomotor stimulating effects of opiates, psychostimulants and NMDA antagonists. The extent to which these effects can be attributed to changes in drug pharmacokinetics and bioavailability versus sensitivity of the neuronal circuits that mediate the affected behavioral functions, has not been established. Recent studies point to central adaptive changes insofar as rewarding, locomotor and c fos-inducing effects of amphetamine and MK-801, injected directly into the lateral ventricle, are greater in food-restricted than ad libitum fed rats. The increased expression of c-fos in nucleus accumbens (NAC) shell, in particular, suggests that food restriction may augment drug reward by modulating dopamine (DA) synaptic function in this area. OBJECTIVES: The first purpose of this study was to investigate whether the rewarding effects of cocaine and the delta1 opioid agonist DPDPE, both of which increase DA synaptic transmission, are augmented by food restriction. The second purpose was to determine whether the delta2 opioid agonist, deltorphin-II, which has been reported to exert DA-independent rewarding effects, is subject to the potentiating effect of food restriction. METHODS: Rewarding effects of drugs were measured in terms of their ability to lower the threshold for lateral hypothalamic self-stimulation (LHSS) using a rate-frequency method. RESULTS: In separate experiments, cocaine (50, 100 and 150 microg, ICV) and DPDPE (10 and 25 microg, ICV) produced greater threshold-lowering effects in food-restricted than ad libitum fed rats. Deltorphin-II (5.0, 10 and 25 microg, ICV) had no effect on reward thresholds, regardless of feeding regimen. CONCLUSIONS: While the reported DA-independence of deltorphin-II rewarding effects seemed to offer a means of testing the hypothesis that DA transmission is the critical modulated variable in food-restricted subjects, rewarding effects of this compound could not be demonstrated in the LHSS paradigm. The present results do, however, confirm and extend prior findings indicating that the enhanced self administration of abused drugs by food-restricted subjects is due to enhanced sensitivity of a final common pathway for drug reward. PMID- 11057525 TI - Enhanced ultrasonic vocalization and Fos protein expression following ethanol withdrawal: effects of flumazenil. AB - RATIONALE: Administration of flumazenil, a benzodiazepine (BZD) antagonist, has therapeutic efficacy against some anxiogenic effects of ethanol withdrawal. This observation has led to the suggestion that anxiety associated with ethanol withdrawal is related to release in brain of an endogenous BZD inverse agonist. OBJECTIVE: The present studies further tested this hypothesis by assessing the effect of flumazenil on withdrawal-induced changes in a behavioral task and on the expression of the neuronal protein, Fos. METHODS: Male Sprague-Dawley rats were withdrawn from a chronic ethanol regimen and tested, with or without flumazenil pretreatment, for either ultrasonic vocalization in response to air puff or for the induction of Fos protein-like immunoreactivity (Fos-LI) in brain. In addition, flumazenil effects on Fos-LI were measured in a group of animals treated with the BZD inverse agonist DMCM (0.75 and 1.0 mg/kg). RESULTS: Flumazenil (5.0 mg/kg) significantly reduced the number of ultrasonic vocalizations observed following withdrawal from chronic ethanol. In contrast, flumazenil (5.0 mg/kg), given either 14 h before withdrawal from chronic ethanol, or during hours 3 and 5 following withdrawal, did not attenuate the effects of withdrawal on Fos-LI. Subsequent testing with DMCM confirmed that a benzodiazepine inverse agonist can induce Fos-LI in most of the same brain regions as observed following ethanol withdrawal, and that this change in Fos protein can be attenuated by pretreatment with flumazenil (5.0 mg/kg). CONCLUSIONS: Overall, these results demonstrate that specific behavioral indices of anxiety, but not measures of Fos-LI, support the contribution of an endogenous BZD inverse agonist in the ethanol withdrawal syndrome. PMID- 11057526 TI - Scopolamine inhibits cocaine-conditioned but not unconditioned stimulant effects in mice. AB - RATIONALE: In animal models, cocaine cues contribute to the development of conditioned responses to the psychomotor stimulating and rewarding effects of the drug. OBJECTIVES: In the present study we investigated the effect of scopolamine, known to impair learning and memory, on cocaine-induced conditioned and unconditioned responses in Swiss Webster mice. METHODS: In the first experiment, mice were treated with saline/saline, saline/cocaine (20 mg/kg), scopolamine (1.0 mg/kg)/cocaine, or scopolamine/saline for 5 days. The treatments were paired with the locomotor activity test cage twice, on days 1 and 5. This allowed to determine: (a) the induction and expression of place-dependent sensitization (PDS) to the psychomotor-stimulating effect of cocaine and (b) place-dependent hyperlocomotion (PDH; i.e., conditioning) as defined by the response to saline injection in the test cage. In the second experiment, all injections were delivered in animals' home cage in order to induce place-independent sensitization (PIS) to cocaine and to avoid the development of PDH. In the third experiment, the effect of scopolamine (1.0 mg/kg) on the acquisition of cocaine induced conditioned place preference (CPP) was investigated. RESULTS: Data from the first experiment suggest that pretreatment with scopolamine had no specific effect on the induction and expression of cocaine-induced PIS. However, scopolamine blocked cocaine-induced PDH. Results from the second experiment confirmed that scopolamine had no effect on the induction of PIS to cocaine. Results from the third experiment showed that scopolamine completely blocked cocaine-induced CPP. CONCLUSIONS: The finding that scopolamine blocked the conditioned behaviors, PDH and CPP, that develop after exposure to cocaine supports the hypothesis that cocaine cue reactivity in the paradigms tested is associated with learning and memory. PMID- 11057527 TI - Anticonflict effect of the glycineB receptor partial agonist, D-cycloserine, in rats. Pharmacological analysis. AB - RATIONALE: Several studies have provided evidence that antagonists and partial agonists of glycine(B) receptors exhibit an anxiolytic-like activity in different animal models. OBJECTIVE: Using the conflict-drinking Vogel test in rats as a model, in the present study we examined the anxiolytic-like activity of D cycloserine (DCS), a partial agonist of the glycine(B) site of the N-methyl-D aspartate (NMDA) receptor complex. Diazepam was used as a reference drug. RESULTS: DCS (200 and 300 mg/kg) and diazepam (5 mg/kg) produced an anxiolytic like effect in rats by increasing the number of shocks accepted. We also demonstrated that NMDA (15 mg/kg) reduced the anxiolytic-like activity of DCS (200 mg/kg), whereas glycine (800 mg/kg) and flumazenil (10 mg/kg) did not affect the anticonflict effect of DCS (200 mg/kg). The anticonflict effect of diazepam (5 mg/kg) was totally blocked by flumazenil (10 mg/kg). CONCLUSION: The obtained results have shown that DCS exhibits an anxiolytic-like activity which depends on NMDA receptors rather than on glycine(B) or benzodiazepine sites. PMID- 11057528 TI - Immaterial rights in medical research. PMID- 11057529 TI - Spinal fusions: bone and bone substitutes. AB - Vertebral arthrodesis is one of the most commonly performed, yet incompletely understood, procedures in spinal surgery. Despite major progress in internal fixation techniques, the high rate of nonunions indicates that physiologic, biologic and molecular events that are crucial to this process are not well known. This article will analyze the general biology of bone regeneration, and particularly discuss the properties and use of various bone graft materials and graft substitutes. PMID- 11057530 TI - Range of global motion of the cervical spine: intraindividual reliability and the influence of measurement device. AB - Range of motion tests are often employed in the quantification of musculoskeletal impairment and in the assessment of the efficacy of therapeutic interventions. The aim of the present study was to compare the absolute values for, and the day to-day reliability of, measures of cervical spinal mobility made with two computerised motion analysis devices. The ranges of cervical flexion, extension, lateral bending, axial rotation, and axial rotation in flexion and extension were determined for 19 volunteers using both the CA6000 Spine Motion Analyser and the Zebris CMS system; all measures were repeated on a second occasion 1-3 days later. The test-retest reliability was good for each instrument: there was no significant difference between the mean values derived on the two separate days (P>0.05), and the corresponding intraclass correlation coefficients were 0.75 0.93 for all pri-O mary movements and 0.57-0.93 for axial rotation in flexion or in extension. For each primary movement, a small but significant difference (1 10%; P<0.05) between the values derived from the two instruments was observed, the systematic nature of which was revealed by the excellent correlation coefficients between them. For the measures of axial rotation in flexion or in extension, however, there was not only a poor correlation between the data obtained from the two devices, but the mean values also differed significantly. Each device is highly reliable in itself and can be used with confidence in longitudinal studies. The establishment of 'normal' values for the primary motions should take account of the slight differences observed between devices. Normal values for rotation in flexion or extension cannot be established until the source of the device-dependent difference is identified. PMID- 11057531 TI - Combined injuries in the upper cervical spine: clinical and epidemiological data over a 14-year period. AB - Concomitant traumatic injuries in the upper cervical spine are often encountered and rarely reported. We examined the data concerning 784 patients with cervical spine injuries following trauma, including 116 patients with upper cervical spine injuries. Twenty-six percent of patients with upper cervical spine injuries (31 cases) were found to have combined injuries involving either the upper or the upper and lower cervical spine. The frequent patterns were combined type I bipedicular fracture of the axis and dens fracture, and combined dens fracture and fracture of the posterior arch of C 1. Other patterns posed specific problems, such as combined dens and Jefferson fracture and combined dens and C2 articular pillar fracture. Seventy percent of atlas fractures, 30% of C2 traumatic spondylolistheses and 30% of dens fractures were part of a combination. A total of 1.7% of patients with lower cervical spine injuries had a combined injury in the upper cervical spine. A comprehensive therapeutic schedule is outlined. Combined injuries in the upper cervical spine should be sought in any patient with a cervical spine injury. PMID- 11057532 TI - CT-guided internal fixation of a hangman's fracture. AB - Most hangman's fractures are treated conservatively. If surgery is indicated, an anterior approach using a C2/C3 graft and plate fusion is usually preferred. Another surgical method according to Judet is direct transpedicular osteosynthesis by the dorsal approach. This surgery is frequently rejected because of the high risk of spinal cord damage or vertebral artery tear. Direct transpedicular osteosynthesis of hangman's fracture according to Judet is a "physiological operation" that does not cause fusion and creates anatomical conditions. This procedure enables appropriate reduction, compression of fragments and immediate stabilization of the C2 segment. A new aspect of Judet's method of internal fixation of a hangman's fracture is now proposed. Computed tomographic (CT) guidance is used to ensure safe and exact introduction of two screws from the posterior approach. This method of CT-guided internal fixation of hangman's fracture allows, preoperatively, for an accurate assessment of the pattern and course of fracture line, selection of the anatomically safest screw path and determination of an appropriate screw length. The procedure also allows for accurate intraoperative control of instrument and implant placement, screw tightening, fracture reduction and anchoring of the screw tip in the contralateral cortex, using repeated CT scans. The procedure is performed in a CT unit under sterile conditions. This method was used in the treatment of eight male and two female patients aged 21-71 years. All treated patients were without neurological deficit. Follow-up ranged from 12 to 57 months (mean 33.3 months). No intraoperative or early or late postoperative complications were apparent. This new aspect of the surgical procedure ensures highly accurate screw placement and minimal risks, and fully achieves the "physiological" internal fixation. PMID- 11057533 TI - A meta-analysis of autograft versus allograft in anterior cervical fusion. AB - We performed a metaanalysis of one- and two-level anterior cervical interbody fusion (ACDF) on data derived from published, peer-reviewed journal articles to determine whether there is a difference in fusion rate, graft complications, or clinical outcome in patients undergoing ACDF according to whether autograft or allograft was used. ACDF is a common procedure for cervical spondylotic radiculopathy. Most published studies comparing autograft and allograft have not demonstrated any difference between grafts. The medical literature dating from 1955 was reviewed. Of 395 titles, only four studies comparing autograft with allograft in ACDF were appropriate for this analysis. The data from these studies -310 patients and 379 intervertebral levels were pooled and statistical methods were applied. For both one- and two-level ACDF, autograft demonstrated a higher rate of radiographic union and a lower incidence of graft collapse. It was not possible to ascertain whether autograft is clinically superior to allograft. Although autograft has a higher fusion rate than allograft, clinical results do not depend solely on radiographic results. The risk of graft site morbidity and patient preference should be considered when choosing the type of graft for this operation. PMID- 11057534 TI - Atlantoaxial immobilization in rheumatoid arthritis: a prophylactic procedure? AB - Timing of surgical intervention in atlantoaxial instability due to rheumatoid arthritis is still controversial. An aim of this study was to investigate whether atlantoaxial fusion can prevent progression of instability and upward migration of the dens. Thirty-two patients with rheumatoid arthritis, who underwent posterior atlantoaxial fixation due to instability, were clinically and radiologically examined after a minimum follow-up of 5 years. The radiological measurements focussed on the extent of cranial vertical migration after atlantoaxial fusion. In none of the 20 patients available for follow-up examination was a vertical cranial migration observed, in spite of the ongoing course of the disease. These findings are in concordance with findings in the literature, and strongly suggest that, with atlantoaxial stabilization, the inflammatory process with destruction of the lateral masses of the atlas is able to prevent further deterioration with vertical cranial migration. PMID- 11057535 TI - Primary stabilizing effect of interbody fusion devices for the cervical spine: an in vitro comparison between three different cage types and bone cement. AB - Interbody fusion cages are small hollow implants that are inserted into the intervertebral space to restore physiological disc height and to allow bony fusion. They sometimes cause clinical complications due to instability, subsidence or dislocation. These are basic biomechanical parameters, which influence strongly the quality of a fusion device; however, only few data about these parameters are available. Therefore, the purpose of the present study was to investigate the primary stabilizing effect of four different cervical fusion devices in in vitro flexibility tests. Twenty-four human cervical spine segments were used in this study. After anterior discectomy, fusion was performed either with a WING cage (Medinorm AG, Germany), a BAK/C cage (Sulzer SpineTech, USA), an AcroMed cervical I/F cage (DePuy AcroMed International, UK) or bone cement (Sulzer, Switzerland). All specimens were tested in a spine tester in the intact condition and after implantation of one of the four devices. Alternating sequences of pure lateral bending, flexion-extension and axial rotation moments (+/- 2.5 Nm) were applied continuously and the motions in each segment were measured simultaneously. In general, all tested implants had a stabilizing effect. This was most obvious in lateral bending, where the range of motion was between 0.29 (AcroMed cage) and 0.62 (BAK/C cage) with respect to the intact specimen (= 1.00). In lateral bending, flexion and axial rotation, the AcroMed cervical I/F cages had the highest stabilizing effect, followed by bone cement, WING cages and BAK/C cages. In extension, specimens fused with bone cement were most stable. With respect to the primary stabilizing effect, cages, especially the AcroMed I/F cage but also the WING cage and to a minor extent the BAK/C cage, seem to be a good alternative to bone cement in cervical interbody fusion. Other characteristics, such as the effect of implant design on subsidence tendency and the promotion of bone ingrowth, have to be determined in further studies. PMID- 11057536 TI - Biomechanical evaluation of a new modular rod-screw implant system for posterior instrumentation of the occipito-cervical spine: in-vitro comparison with two established implant systems. AB - Posterior instrumentation of the occipito-cervical spine has become an established procedure in a variety of indications. The use of rod-screw systems improved posterior instrumentation as it allows optimal screw positioning adapted to the individual anatomic situation. However, there are still some drawbacks concerning the different implant designs. Therefore, a new modular rod-screw implant system has been developed to overcome some of the drawbacks of established systems. The aim of this study was to evaluate whether posterior internal fixation of the occipito-cervical spine with the new implant system improves primary biomechanical stability. Three different internal fixation systems were compared in this study: the CerviFix System, the Olerud Cervical Rod Spinal System and the newly developed Neon Occipito Cervical System. Eight human cervical spine CO/C5 specimens were instrumented from C0 to C4 with occipital fixation, transarticular screws in C1/C2 and lateral mass or pedicle screws in C3 and C4. The specimens were tested in flexion/extension, axial rotation, and lateral bending using pure moments of +/- 2.5 Nm without axial preload. After testing the intact spine, the different instrumentations were tested after destabilising C0/C2 and C3/C4. Primary stability was significantly increased, in all load cases, with the new modular implant system compared to the other implant systems. Pedicle screw instrumentation tended to be more stable compared to lateral mass screws; nevertheless, significant differences were observed only for lateral bending. As the experimental design precluded any cyclic testing, the data represent only the primary stability of the implants. In summary, this study showed that posterior instrumentation of the cervical spine using the new Neon Occipito Cervical System improves primary biomechanical stability compared to the CerviFix System and the Olerud Cervical Rod Spinal System. PMID- 11057537 TI - Osteoid osteoma of the cervical spine depicted as dumbbell tumor by MRI. AB - We report a case of 14-year-old male patient with osteoid osteoma of the cervical spine. Magnetic resonance imaging (MRI) revealed a large dumbbell-shaped paravertebral tumor in the region of the exiting left C6 nerve. A computed tomographic (CT) scan after myelography showed a much smaller bony defect in the medial aspect of the left C6 pedicle with central calcification and extensive bone sclerosis around the defect, typical of osteoid osteoma. The diagnosis was confirmed postoperatively. The resected specimen exhibited extensive vascularization of the osteoid tissue. The case is presented because MRI did not allow a specific diagnosis of osteoid osteoma, and suggested the tumor was larger than in reality it was, by also depicting the reactive inflammation around the tumor as if it were part of the tumor. PMID- 11057538 TI - Traumatic vertical atlantoaxial instability: the risk associated with skull traction. Case report and literature review. AB - Traumatic overdistraction between C1 and C2 may occur when all the ligaments connecting C2 to the skull are ruptured, and may be manifested when an attempt to reduce C1-C2 subluxation is made by means of traction. We describe here the case of a patient with traumatic anterior atlantoaxial dislocation, who developed atlantoaxial vertical dissociation after skull traction using a Gardner-Halo with lb 4.02 (1.5 kg) of weight. The identification of patients who are susceptible to this complication is difficult. In this case, it might have been prevented by avoiding spinal traction. The aim of this report was to show that vertical dissociation may occur in C -C2 anterior dislocation submitted to spinal traction, and that other forms of reduction must be considered to treat these pathologies and avoid this potentially fatal complication. PMID- 11057539 TI - Haemangioblastoma of a cervical sensory nerve root in Von Hippel-Lindau syndrome. AB - Spinal haemangioblastomas are rare, accounting for only about 7% of all central nervous system cases. The case of a 40-year-old woman with a haemangioblastoma arising solely from a cervical sensory nerve root is presented. At operation via a cervical laminectomy, it was possible to resect the tumour en masse with the sensory ramus, by extending the laminectomy through the exit foramen for C6. Haemangioblastomas are commonly intramedullary, and have only been reported in this location on one previous occasion. The patient has Von Hippel-Lindau syndrome and a history of multiple solid tumours. The possible role of the Von Hippel-Lindau tumour suppressor gene in the pathogenesis of these neoplasms is discussed. PMID- 11057540 TI - Vertebral body replacement with a bioglass-polyurethane composite in spine metastases--clinical, radiological and biomechanical results. AB - Metastatic spine lesions frequently require corpectomy in order to achieve decompression of the spinal cord and restoration of spinal stability. A variety of systems have been developed for vertebral body replacement. In patients with prolonged life expectancy due to an improvement of both systemic and local therapy, treatment results can be impaired by a loosening at the implant-bone interface or mechanical failure. Furthermore, early detection of a metastatic recurrence using sensitive imaging modalities like computed tomography (CT) and magnetic resonance imaging (MRI) is possible in these patients without artefact interference. The aim of our pilot study was to evaluate the clinical applicability and results of a new radiolucent system for vertebral body replacement in the lumbar spine. The system consists of bone-integrating biocompatible materials - a polyetherurethane/bioglass composite (PU-C) replacement body and an integrated plate of carbon-fibre reinforced polyetheretherketone (CF-PEEK) - and provides high primary stability with anterior instrumentation alone. In a current prospective study, five patients with metastatic lesions of the lumbar spine were treated by corpectomy and reconstruction using this new system. Good primary stability was achieved in all cases. Follow-up (median 15 months) using CT and MRI revealed progressive osseous integration of the PU-C spacer in four patients surviving more than 6 months. Results obtained from imaging methods were confirmed following autopsy by biomechanical investigation of an explanted device. From these data, it can be concluded that implantation of the new radiolucent system provides sufficient long-term stability for the requirements of selected tumour patients with improved prognosis. PMID- 11057541 TI - Percutaneous transpedicular vertebroplasty with PMMA: operative technique and early results. A prospective study for the treatment of osteoporotic compression fractures. AB - Vertebroplasty-percutaneous cement augmentation of osteoporotic vertebrae is an efficient procedure for the treatment of painful vertebral fractures. From a prospectively monitored series of 70 patients with 193 augmented vertebrae for osteoporotic and metastatic lesions, we analysed a group of 17 patients suffering from back pain due to osteoporotic fractures. The reinforcement of 45 vertebral bodies in these patients led to a significant and lasting pain reduction (P < 0.01 ). The presented technique is useful, as, in one session, at least four injections can be performed when required, allowing the prophylactic reinforcement of adjacent vertebrae as well. The use of a low-viscosity polymethyl methacrylate (PMMA) in combination with a non-ionic liquid contrast dye provides a reliable and safe procedure. Extraosseous cement leakage was seen in 20% of the interventions; however, none of them had clinical sequelae. PMID- 11057542 TI - A simplified Galveston technique for the stabilisation of pathological fractures of the sacrum. AB - Mechanical stabilisation of pathological fractures of the sacrum is technically challenging. There is often inadequate purchase in the sacrum, and stabilisation has to be achieved between the lumbar vertebrae and ilium. We present a simplification of the Galveston technique. We treated a total of six patients with this technique, four for metastatic disease and two for primary tumours. Our technique consists of the formation of a proximal stable construct using ISOLA pedicle screws linked distally using a modular system of connectors to threaded iliac bolts with cross linkages. Neurological decompression and fusion was performed as appropriate. The benefits of this method are: ease of access to the ilium, a solid purchase to the ilium, less rod contouring and shorter operating time. We have had no operative complications from this procedure. All patients were discharged home mobile, with a reduced opiate requirement. PMID- 11057543 TI - The contribution of the adriamycin-induced rat model of the VATER association to our understanding of congenital abnormalities and their embryogenesis. AB - The adriamycin-induced rat model of the VATER association has provided a means of studying the morphogenesis of a variety of major congenital structural abnormalities similar to those seen in humans with the VATER association. Most interest has been centered on the foregut, where the model has clarified some aspects of the development of esophageal atresia (EA), tracheal agenesis, and other communicating bronchopulmonary foregut malformations. It has demonstrated aberrations in the nerve supply to the esophagus in EA and allowed the study of tracheomalacia. A relationship between an abnormal notochord, foregut abnormalities, and vertebral defects has been shown, and the model has reignited interest in the role of the notochord as a regional organizer of axial development. The normal temporospatial characteristics of apoptosis during fore- and hindgut development is disturbed in this model, resulting in abnormal morphology. The indications are that this model will continue to clarify the processes that lead to many of the structural congenital abnormalities that are seen in infants born with the VATER association. PMID- 11057544 TI - Effect of hyperoxia on surfactant protein gene expression in hypoplastic lung in nitrofen-induced diaphragmatic hernia in rats. AB - The hypoplastic lung in congenital diaphragmatic hernia (CDH) has both a quantitative and qualitative reduction in surfactant. Recently, the role of oxygen (O2) as a regulator of pulmonary surfactant-associated protein (SP) gene expression has been reported. The mRNA level of SP has been demonstrated to be increased in the lungs of animals exposed to hyperoxia. The aim of this study was to investigate SP mRNA expression in hypoplastic CDH lung in rats during mechanical ventilation in order to determine the effect of O2 on SP synthesis in CDH. A CDH model was induced in pregnant rats following administration of nitrofen. The newborn rats with CDH and controls were intubated and ventilated. Ventilation was continued for 6 h under 100% oxygen. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to evaluate the relative amounts of mRNA expression of SP-A, SP-B, SP-C, and SP-D. Relative amounts of SP-A, SP-B, and SP-D mRNA expression in CDH lung were significantly decreased compared to controls at birth and 6 h after ventilation. There was no significant difference in SP-C mRNA expression between CDH animals and controls. Upregulated mRNA expression of SP-A, SP-B, and SP-D in lungs of control animals at 6 h after ventilation suggests that oxygenation accelerates postnatal SP synthesis in normal lungs. The inability of O2 to increase SP mRNA expression in hypoplastic CDH lung suggests that the hypoplastic lung is not responsive to increased oxygenation for the synthesis of SP. PMID- 11057545 TI - Esophageal atresia and tracheoesophageal fistula: a review of 25 years' experience. AB - During a 25-year period (1972-1996), 585 patients with esophageal atresia with or without tracheoesophageal fistula were treated at the Department of Pediatric Surgery, SMS Medical College, Jaipur, India. Increasing awareness of the anomaly has led to early detection and referral with fewer pulmonary complications. For purposes of analysis the period has been divided into five phases, with a steady decline in overall mortality observed from 95.4% in phase 1 to 41% in phase V. Although postoperative complications have also shown a declining trend, delay in diagnosis, prematurity, low birth weight, delayed arrival at the surgical centers, sepsis, pulmonary complications including pneumonitis, and inadequate nursing care all continue to contribute substantially to lower the survival in developing countries such as ours. PMID- 11057546 TI - Corrosive injuries of the esophagus in newborns. AB - To evaluate the morbidity and mortality of corrosive esophageal injuries (CEI) in the neonatal period, the records of 184 children hospitalized following caustic ingestion over a 10-year period from January 1987 to November 1997 were reviewed. Eight (4.3%) were newborns (5 boys and 3 girls). The mean age of the newborns was 12 days (range 1-28). The ingested caustic materials were benzalkonium chloride in six patients and trichloroacetic acid in two. Oropharyngeal examination and esophagoscopy were performed for diagnosis. Hyperemia and fibrin plaques were present in the oropharynx in all patients. The management consisted of endotracheal intubation, antibiotics, corticosteroids, and total parenteral nutrition. Pneumonia and sepsis developed in three patients and one died of sepsis. Stenosis developed in two patients, who were treated three times with antegrade dilatations. The morbidity was 62.5% (five patients) and the mortality was 12.5% (one) in newborns with CEI. These results indicate that ingestion of a caustic substance results in high morbidity and mortality in newborns. Parents and nurses should be warned about this risk. PMID- 11057547 TI - Apoptosis in murine duodenum during embryonic development. AB - Duodenum is thought to go through a solid-core stage followed by recanalization during its development. This study investigates the role of apoptosis in normal duodenal development, especially during widening of the lumen, and hence, the possible role of apoptosis in duodenal atresia (DA). Twenty-four time-mated Sprague-Dawley rats were killed from day 13 to day 20 of gestation. Duodenums of 3 fetuses were chosen randomly from each rat and processed. Apoptosis was determined by the terminal deoxytransferase-mediated biotin dUTP nick-end labeling (TUNEL) technique (ApopTag). Apoptosis count and cross-sectional areas were measured with an image analyzer (MetaMorph). The number of apoptotic cells per unit area duodenum peaked on day 15 for the mucosal/submucosal layer and on day 14 for the muscular/mesenchymal layer. The maximal number of apoptotic cells per cross-section of duodenum was between 7 and 8. The cross-sectional areas of the duodenal wall and lumen increased exponentially between day 17 and day 19 while duodenal-wall thickness remained relatively constant throughout duodenal development. The localization, timing, and intensity of apoptosis do not suggest that apoptosis is responsible for the widening of the duodenal lumen; enlargement of the lumen is related to the increase in duodenal circumference. Apoptosis thus may not be involved in the pathogenesis of DA. PMID- 11057548 TI - Infantile acute pancreatitis after mumps vaccination simulating an acute abdomen. AB - We describe an extremely rare case of acute pancreatitis presenting as an acute abdomen that appeared as a complication of mumps vaccination in a young child. A laparotomy performed because of suspected perforated appendicitis proved unnecessary in retrospect. No similar case in infancy and early childhood has been reported to date. PMID- 11057549 TI - The effect of the availability of laparoscopic techniques on the treatment of appendicitis in children. AB - The use of laparoscopic surgery for acute appendicitis (AA) in children has increased over recent years. The aim of this study was to determine what effect the availability of laparoscopic appendicectomy (LA) has had on children admitted with suspected AA. A retrospective review of children admitted between January 1994 and June 1999 inclusive who underwent appendicectomy for suspected AA was conducted. Data recorded included standard demographic information, surgical approach, histopathology of the appendix, complications, and post-operative length of stay. After LA, children had a shorter post-operative stay, although the technique was used less frequently in advanced disease. The rate of normal histology was higher for LA. Laparoscopic surgery was performed in females in 69% of whom 48% had a normal appendix removed. The duration of surgery was longer for LA (59 vs 40 min). The rate of LA increased to 77% in the first 6 months of 1999. There was a decrease in the proportion of laparoscopic procedures converted to open appendicectomy from 50 to 6% during the period reviewed. The ready availability of laparoscopy and increased confidence in its use has resulted in more children, especially females, with suspected AA undergoing laparoscopy. There was a corresponding higher rate of normal appendix removal in this group, but the overall rate of normal histology has not changed, suggesting that the laparoscopic approach is more likely to be employed where the clinical diagnosis is less certain, particularly in older girls. PMID- 11057550 TI - Long-term results of treatment of single-system ectopic ureters. AB - Single-system ureteral ectopia (UE) encompasses a spectrum of malformations involving the bladder trigone, ureter, and kidney. The clinical presentation is variable, and both diagnostic and therapeutic problems are common. Reduced renal function in these patients may result from primary dysplasia, obstruction, vesicoureteral reflux, or recurrent infection. Based on our experience of seven patients, suggestions for diagnostic procedures and criteria for renal saving versus nephrectomy are offered. The relationship between ostium localization, renal function, and long-term results was investigated. From 1972 to 1990, five female and two male patients were studied. During the same period, 31 patients with UE and duplex kidneys were seen. Ages ranged from 1 day to 7 years. A ureteric opening into the bladder neck was associated with dilatation of the ureter and renal pelvis. Two patients had vaginal ectopia and severe renal dysplasia. In one, a cyst of the vaginal wall (Gardner's cyst) was detected at birth. A male newborn had multicystic renal dysplasia on the left and ureteric ectopia to the ductus deferens on the right side. To our knowledge, he is the first patient reported with renal function totally dependent on a kidney with severe UE. Follow-up ranged from 4 to 9 years. One patient died in the postoperative period because of renal failure and sepsis. All the others are well and have normal creatinine values. Improvement of renal function was noted after ureteral reimplantation (URI) in patients with bladder-neck ectopia. The numbers of infections were also drastically reduced. Our observations suggest that the combination of ultrasound, cyst urethrography, and cystoscopy will be diagnostic in most patients. A suspicion of UE should be raised in symptomatic patients with apparently solitary kidneys, enuresis ureterica, or atypical obstructive uropathy. Reduced renal function in some patients with ectopia to the bladder neck will improve after URI. This may be of importance in patients with bilateral anomalies and marginal renal function. PMID- 11057551 TI - Comparative studies of fertility and histologic development of contralateral scrotal testes in two rat models of unilateral cryptorchidism. AB - Fertility and the development of the contralateral scrotal testis in patients with unilateral cryptorchidism (UCO) remain controversial. This study investigated these controversies in two different UCO rat models using 43 Wistar King A rats. The animals were divided into three groups: I: an endocrinologic model of UCO was obtained by injecting pregnant dams with flutamide 100 mg/kg per day on days 15-17 of gestation (n = 12); II (n = 21): a mechanical model of UCO was obtained by extra-abdominal fixation of the gubernaculum in the neonatal period, III (n = 10): non-treated rats were used as controls. At the age of 90 days, 5 rats from each group were segregated into individual cages and housed with two virgin adult females for 2 weeks. The occurrence of pregnancy and litter sizes were counted in order to study fertility. All the animals were then weighed and killed. The occurrence of testicular descent, growth of the external genitalia, and epididymal development were examined. Morphologic and histologic evaluations were performed in the cryptorchid and contralateral testes. In the endocrinologic model (group I) the 10 female rats failed to show any offspring (0%), while in the mechanical model (group II) 9 out of 10 rats had offspring (90%, P < 0.001); 10 out of 10 control rats showed offspring. All of the rats in groups I and II had UCO, and the undescended testes were located in the superficial inguinal position, while the contralateral and control testes descended into the scrotum. Hypospadias and a small epididymis were frequently noted in the flutamide-treated rats. Testicular weight, seminiferous tubular diameter, and spermatogenesis were all significantly reduced in the undescended testes (UDT) compared to the contralateral and control testes. Moreover, the development of the contralateral testis was inhibited in group I compared to groups II and III. Our observations showed that short-term exposure to flutamide in utero induced significantly reduced fertility and degenerated contralateral scrotal testes in UCO rats compared to mechanically-induced UCO rats by early adulthood. It is suggested that fertility potential and testicular development in unilateral UDT may be partially due to the factors that induce testicular maldescent, especially in cases due to intrauterine hormonal abnormalities. These cases may show inhibited fertility and testicular development even after orchiopexy. PMID- 11057552 TI - Effects of male sex hormones on urodynamics in childhood: intersex patients are a natural model. AB - The effects of sex hormones on bladder function have been evaluated in adult females, especially in regard to postmenopausal incontinence and bladder irritability syndromes. These have not been investigated in children in regard to urodynamic findings. An intersex patient whose bladder is under the influence of androgens is a natural model to investigate the effects of male sex hormones on bladder function in females. To evaluate the urodynamic findings and clinical symptoms in a group of intersex patients and to determine how androgens influence bladder function in female children, clinical and urodynamic records of 12 intersex patients with adrenogenital syndrome were investigated retrospectively. The mean age was 9 +/- 5.7 years (1.5-18) and the mean follow-up period was 5.1 +/- 4.4 years (1-12). Congenital adrenal hyperplasia (CAH) was present in all cases. Only 3 patients had urinary symptoms and incontinence, but these findings did not correlate with their urodynamic findings. None of the patients required medications for their urinary symptoms. Nine are still being treated medically by the pediatric endocrine team with hydrocortisone for CAH. The upper urinary tract was found to be normal with no hydronephrosis. The mean bladder capacity (269 +/- 122 ml) was lower (86.7%) than the estimated capacity for age. The mean compliance was 20 +/- 13.7 ml/cmH2O. No unstable detrusor contractions were encountered. The most remarkable finding was this reduced bladder capacity of androgenized female patients for age, which shows the antagonistic effect of androgens on bladder urodynamics in females. PMID- 11057553 TI - Blunt abdominal trauma in children: epidemiology, management, and management problems in a developing country. AB - Trauma is the leading cause of death in children in developed countries. In tropical Africa, it is only beginning to assume importance as infections and malnutrition are controlled. In developed countries, the availability of advanced imaging modalities has now reduced the necessity for laparotomy to less than 10% following blunt abdominal trauma (BAT) in children. This report reviews the epidemiology, management, and unnecessary laparotomies for pediatric BAT in a developing country in a retrospective review of 57 children aged 15 years or less at the Ahmadu Bello University Teaching Hospital, Zaria, Nigeria over 12 years. The average age was 9 years and the male-female ratio 3.8:1. Seventy-four percent (74%) of abdominal injuries in children were due to blunt trauma. The commonest causes of injury were road traffic accidents (RTA) (57%), 88% in pedestrians and 59% in children aged 5-9 years. Falls were the cause of trauma in 36%, 60% of them aged 10-15 years. Other causes of injury were sports in 5% and animals in 2%. Diagnosis was clinical, supported by diagnostic peritoneal lavage or paracentesis. Two patients had ultrasonography, and none had computed tomography. Fifty-three patients had a laparotomy, 2 died before surgery, 1 was managed nonoperatively, and in 1 surgery was declined. There were 34 splenic injuries, 20 treated by splenic preservation, splenectomy in 13, and non-operative in 1. Fourteen gastrointestinal injuries were treated in 12 patients. Of 9 hepatic injuries, 4 were minor and were left untreated, 3 were repaired, 1 was packed to arrest hemorrhage, and a lacerated accessory liver was excised. Four injuries to the urinary tract (bladder contusion 2, bladder rupture 1, ruptured hydronephrotic kidney 1) were treated accordingly. There were 4 retroperitoneal hematomas associated with other intra-abdominal injuries and 2 pancreatic contusions. One lacerated gallbladder was treated by cholecystectomy and a ruptured left hemidiaphragm was repaired transperitoneally. In retrospect, 27 (51%) patients could have been managed by observation (splenic injury 20, liver injury 5, bladder contusion 2) using advanced imaging modalities. One patient developed an intra-abdominal abscess following splenorrhaphy. The average hospital stay was 17 days. Mortality was 8 (14.5%) from gastric perforation (3), liver injury (2), splenic injury (1), and 2 patients died before surgery. BAT in this population results predominantly from RTA in pedestrians. Laparotomy may be avoided in 51% of cases if advanced imaging modalities are readily available. PMID- 11057554 TI - Salivary fistulae. AB - Three cases of unusual cervical fistulae are presented with a review of the literature to caution against labelling all lateral cervical fistulae as simple branchial fistulae or midline ones as dermoid cysts. PMID- 11057555 TI - Congenital diaphragmatic hernia in identical twins. AB - The authors present a pair of identical twins with congenital diaphragmatic hernia (CDH) diagnosed prenatally, who underwent successful surgical repair. They were diagnosed as having CDH at 32 weeks' gestation and showed respiratory distress soon after cesarean section at 33 weeks' gestation. Both survived after scheduled perinatal management followed by surgery, for which the prenatal diagnosis of CDH was valuable. PMID- 11057556 TI - Hepatic lymphangioma--a case report. AB - A patient presented with a huge, pedunculated abdominal cystic lymphangioma arising from the quadrate lobe of the liver near the round ligament. Microscopically, dilated hepatic ducts with scant liver tissue could be recognized in the main cyst. A review of the literature reveals no previous report of a lymphangioma arising in this manner or from this area. PMID- 11057557 TI - Relapsing pancreatitis in a child duplication in an aberrant pancreatic lobe. AB - An aberrant pancreatic lobe associated with an enteric duplication cyst is a rare cause of relapsing pancreatitis in childhood. We present an 8-year-old boy with relapsing pancreatitis caused by this rare congenital foregut anomaly. The computed tomography (CT) findings revealed an unusually long segment of aberrant pancreatic lobe arising from the pancreatic neck, projecting anteriorly at a distance to a cystic duodenal duplication and appearing as an inflammatory mass. There has been no previous report of this unusual appearance on CT. Appreciation of the relevant anatomy provided by CT led to the successful management of this surgically-treatable cause of relapsing pancreatitis. PMID- 11057558 TI - Repair of a giant omphalocele by a modified technique. AB - Large omphaloceles that contain centrally herniated liver pose challenges to surgical closure, the most significant being the space limitation of the abdominal cavity. In addition, the "pedicled" nature of the liver on the inferior vena cava creates a predisposition to acute hepatic vascular outflow obstruction as the liver is reduced into the abdominal cavity. In such cases, the alternatives include conservative treatment or staged silo reduction. The worst complication of silastic silo (SS) placement is tension and infection of the fascia with disruption of the suture line. Once infection or premature disruption occurs, closure of the defect is difficult or impossible. This case report details a different management technique for a newborn with a giant omphalocele and presents an interesting variation of the usual SS technique that may be helpful in the management of some cases, especially in an emergency. The thick silk sutures applied in the present case absorbed the tension and the silastic sheet prevented the risks of infection and adhesions. PMID- 11057559 TI - Volvulus of the transverse and sigmoid colon. AB - A case of transverse and sigmoid-colon volvulus and a discussion of the probable mechanism of large-bowel volvulus (LBV) in children and its management is presented. A 5-year-old male with cerebral palsy presented with transverse-colon and subsequently sigmoid volvulus. The child underwent resection of the involved segments with primary colocolic and colorectal anastomosis, respectively. The recovery was uneventful. LBV in children is due to congenital anomalous or absent ligamentous fixation of the colon. Constipation is probably the result of the volvulus. Resection of the involved segment and primary anastomosis is the definitive treatment. PMID- 11057560 TI - Large bowel obstruction: an unusual presentation of salmonella enterocolitis in infancy. AB - An infant with a large-bowel obstruction due to Salmonella B enterocolitis is presented. The clinical and radiologic findings were suggestive of Hirschsprung's disease with total colonic aganglionosis. Due to further deterioration, an ileostomy was performed. Pathologic examination disclosed ganglia in the colon. At laparatomy, 1 month later the colon, which was almost completely obliterated, was resected and an ileorectal anastomosis carried out. The patient remained a carrier of a multiple antibiotic-resistant group B Salmonella strain, and 2 months later died as a result of severe gastroenteritis. PMID- 11057561 TI - The egg-shell sign: a possible indicator of raised intrarenal pressure. AB - Prenatal diagnosis of pelvicalyceal dilatation has produced clinical material that we are continually reinterpreting with the help of improving ultrasound equipment. However, the ability to predict the outcome for any one patient with marked dilatation remains poor. We describe a new sign that may help identify those fetuses who have high intrarenal pressure and therefore justify more aggressive management, while obviating the need for intervention for those in whom it is not present. The egg-shell sign consists of a thin crescent of increased echogenicity over a distended calyx and, in this case, was documented to be associated with other features of raised intrarenal pressure. PMID- 11057562 TI - Management of Wilms' tumor with intracardiac extension. AB - We review our experience and the literature in treating 4 patients with Wilms' tumor (WT) with intracardiac extension among 92 patients with this neoplasm. Cardiopulmonary bypass with circulatory arrest and profound hypothermia was used. There were 3 boys (3 years, 4 years 5 months, and 15 years) and 1 girl (6 years). The follow-up periods were 8 months, 3 years, 2 years 6 months, and 15 years, respectively. We had no surgical complications and conclude that the preoperative diagnosis is extremely important. These patients must be transferred to institutions where concomitant cardiac procedures can be performed. In treating patients with WT, Doppler ultrasound must be used preoperatively in all cases, not only those in which clinical and radiologic signs of intravascular involvement are found. We propose that preoperative chemotherapy should be used, as it shrinks the thrombus and causes desirable adherence of the thrombus to the venous wall, reducing the probability of thromboembolism during the surgical procedure. We also find this method safer than in our 1st case, where neither cardiac arrest nor hypothermia was used. Our results agree with the literature that intracardiac extension of WT does not worsen its prognosis when a rational surgical approach is used. PMID- 11057563 TI - Congenital balloon digits in two neonates caused by constriction rings. AB - Balloon digits were found in two neonates with congenital constriction ring syndrome. The affected digits were the right long finger and right great toe. They were surgically treated at the age of 10 and 9 days, respectively. Morphologic improvement was dramatic after surgery. In cases with extensive enlargement, severe cyanosis, redness, and no subsidence of edema within several days after birth, early operative treatment may be necessary to maintain digit viability and prevent autoamputation due to circulatory embarrassment. It can also be helpful to prevent fibrosis of the subcutaneous tissue. Pathologic examination revealed marked proliferation of fibrous tissue and lymphatic vessels. PMID- 11057564 TI - Silo reduction of giant omphalocele and gastroschisis utilizing continucous controlled pressure. AB - A method is described utilizing continuous controlled pressure to achieve smooth, rapid, and safe silo reduction of an anterior abdominal-wall defect. A metal tube with larger wheels at each end is suspended by runners and counterweights to slowly roll the silo and squeeze the contents into the abdominal cavity. PMID- 11057565 TI - A simple method of intraoperative confirmation of intestinal patency. AB - Routine intraoperative rectal temperature monitoring may serve in addition as a means of distal intestinal patency confirmation. A simple method, which is of immense importance especially when operating on infants and small children after NEC or intestinal atresia, is described. PMID- 11057566 TI - Analytical method for the determination of O,O-diethyl phosphate and O,O-diethyl thiophosphate in faecal samples. AB - A residue analytical method was developed for the determination of the dialkylphosphate metabolites of parathion in faecal samples obtained from rabbits. The faecal pieces were homogenised in water and highly water-soluble O,O diethyl phosphate (DEP) and O,O-diethyl thiophosphate (DETP) were subsequently alkylated to pentafluorobenzyl esters by a phase transfer reaction. Derivatisation yields depend on the reaction time. The recovery rates were determined over the complete procedure using authentic reference standards in matrix solution. The reference standards allow to observe an effect of the sample matrix on the area of signals while GC-FPD is used. The recoveries over the concentration range from 0.05 to 5 microg/g were 47-62% for O,O-diethyl phosphate and 92-106% for O,O-diethyl thiophosphate potassium salt with FPD. PMID- 11057567 TI - Influence of oxygen supply on heptadecane mineralization by Pseudomonas nautica. AB - The influence of different states of oxygen supply on heptadecane mineralization has been investigated in resting cell suspensions of Pseudomonas nautica. The rate of heptadecane biodegraded was constant for oxygen concentrations between 21% and 10% (v/v) (about 100% and 50% of air saturation, 230 and 110 microM, respectively). A decline in biodegradation rates occurred for oxygen concentrations below 10% (about 50% of air saturation, 110 microM) and biodegradation stopped with 0.21% oxygen (2.3 microM). In the presence of 1% (11 microM) of oxygen, no CO2 was produced, showing that complete mineralisation of heptadecane was blocked. PMID- 11057568 TI - Influence of soil texture and tillage on herbicide transport. AB - Two long-term no-till corn production studies, representing different soil texture, consistently showed higher leaching of atrazine [2-chloro-4-(ethylamino) 6-(isopropylamino)-s-triazine] to groundwater in a silt loam soil than in a sandy loam soil. A laboratory leaching study was initiated using intact soil cores from the two sites to determine whether the soil texture could account for the observed differences. Six intact soil cores (16 cm dia by 20 cm high) were collected from a four-year old no-till corn plots at each of the two locations (ca. 25 km apart). All cores were mounted in funnels and the saturated hydraulic conductivity (Ksat) was measured. Three cores (from each soil texture) with the lowest Ksat were mixed and repacked. All cores were surface treated with 1.7 kg ai ha(-1) [ring-14C] atrazine, subjected to simulated rainfall at a constant 12 mm h(-1) intensity until nearly 3 pore volume of leachate was collected and analyzed for a total of 14C. On an average, nearly 40% more of atrazine was leached through the intact silt loam than the sandy loam soil cores. For both the intact and repacked cores, the initial atrazine leaching rates were higher in the silt loam than the sandy loam soils, indicating that macropore flow was a more prominent mechanism for atrazine leaching in the silt loam soil. A predominance of macropore flow in the silt loam soil, possibly due to greater aggregate stability, may account for the observed leaching patterns for both field and laboratory studies. PMID- 11057569 TI - Lichens and mosses as biomonitors of trace elements in areas with thermal springs and fumarole activity (Mt. Amiata, central Italy). AB - The contribution of thermal springs and fumaroles to environmental levels of Al, As, B, Cd, Cu, Fe, Hg, Mo, Pb, S, Sb and Zn was evaluated by means of lichens (Parmelia sulcata) and mosses (Hypnum cupressiforme) used as bioaccumulators. Compared to the data reported for unpolluted areas, accumulation of Hg, S and Al was found, with values of Hg and S in the same range as in the vicinity of geothermal power plants. Furthermore, fumaroles turned out to be a significant source of atmospheric arsenic. PMID- 11057570 TI - Estimation of soil adsorption coefficients of organic compounds by HPLC screening using the second generation of the European reference soil set. AB - The European reference soil set was introduced as common basis for a better comparability of soil sorption data measured within the framework of chemical testing of environmental chemicals. The success of the EUROSOILS, as the set is commonly called, convinced the European Commission's Joint Research Centre to evaluate the possibility of producing a remake of these unique and new type of reference materials maintaining the principal sorption-controlling properties of the soils. In this paper the recently proposed second generation of the EUROSOILS is used to evaluate a HPLC-screening technique for the estimation of soil adsorption coefficients of organic chemicals. It could be shown that the derived correlations between HPLC capacity factors of the test substances and the respective soil adsorption coefficients resulting from batch experiments with the second version of the EUROSOILS agreed with those derived for the first generation of reference soils at a different occasion. PMID- 11057571 TI - Comparison of particulate mass, chemical species for urban, suburban and rural areas in central Taiwan, Taichung. AB - Aerosol samples for PM2.5, PM(2.5-10) and TSP were collected from June to September 1998 and from February to March 1999 in central Taiwan. Ion chromatography was used to analyze the acidic anions: sulfate, nitrate and chloride in the Universal samples. The ratios of fine particle concentrations to coarse particle concentrations displayed that the fine particle concentrations are almost greater than that of coarse particle concentrations in Taichung area. The average concentrations of PM2.5, PM(2.5-10) and TSP in urban sites are higher than in suburban and rural sites at both daytime and night-time. Chloride dominated in the coarse mode in daytime and in fine mode in night-time. Nitrate can be found in both the coarse and fine modes. Sulfate dominated in fine mode in both daytime and night-time. PMID- 11057572 TI - Biodegradation of a nonylphenol ethoxylate by the autochthonous microflora in lake water with observations on the influence of light. AB - Alkylphenol polyethoxylates (APE) are routinely used as additives in pesticide formulations. Biodegradation of APEs results in the accumulation of persistent short chain mono-, di- and tri-ethoxylates (AP1EO, AP2EO AP3EO) that are more toxic than the parent compounds and potentially oestrogenic. Accumulation of persistent APE metabolites in shallow or ephemeral waters may pose a hazard to aquatic fauna. This study has followed the degradation and formation of individual oligomers in freshwater in static die-away tests with and without illumination. Over 33 days in darkness there was a progressive and complete loss of long chain oligomers (NP8-17EO), transient increases and subsequent loss of short to medium chain oligomers (NP4-7EO), and large persistent increases (approximately 1000%) in short chain oligomers (NP1-3EO). In the presence of illumination, biodegradation was retarded and heterotrophic bacterial proliferation was inhibited. After 33 days there was complete loss of long chain oligomers (NP9-17EO), incomplete loss of medium chain oligomers (NP6-8EO) and increases in short chain oligomers (NP1-5EO). PMID- 11057573 TI - Concentrations of some heavy metals in water, sediment and fish species from the Ataturk Dam Lake (Euphrates), Turkey. AB - Concentrations of heavy metals (Cd, Co, Cu, Fe, Hg, Mn, Mo, Ni, Pb and Zn) were measured in the water, sediment and fish species (Acanthobrama marmid, Chalcalburnus mossulensis, Chondrostoma regium, Carasobarbus luteus, Capoetta trutta and Cyprinus carpio) from the Ataturk Dam Lake, Turkey. Among the heavy metals studied Cd, Co, Hg, Mo and Pb were not detected in water, sediments and fish samples, while Ni was undetectable levels in fish samples. Levels of Cu, Fe, Mn and Zn varied depending on different tissues. The results of this study indicated that a general absence of serious pollution in the dam lake is due to heavy metals, where as the concentrations of elements found could mainly be attributed to geological sources. PMID- 11057574 TI - Simulating the response of metal contaminated lakes to reductions in atmospheric loading using a modified QWASI model. AB - The changes in metal concentration following significant reductions in atmospheric metal loading of two nickel and copper contaminated lakes in Coniston Valley of the Sudbury Basin of Ontario, Canada were simulated by using steady state and dynamic versions of a modified Quantitative Water Air Sediment Interaction (QWASI) Model. Metal partitioning and precipitation processes were quantified with the aid of US EPA's MINTEQA2 Model. The dynamic model successfully described the recovery of the two lakes and identified key input, loss and partitioning processes. A useful modelling strategy is to develop one or more steady-state models that give an approximate representation of conditions at defined times, then extend this to a dynamic version which can take into account the differing rates of response of components of the system. This modelling strategy can be used for designing and assessing remediation programs for metal contaminated lakes and watersheds. PMID- 11057575 TI - Correlating the physical-chemical properties of phthalate esters using the 'three solubility' approach. AB - A quantitative structure-property relationship (QSPR) method for the correlation of physical-chemical properties and partition coefficients, namely the 'three solubility' approach, is described and applied to a group of 22 phthalate esters. The solubilities or 'apparent-solubilities' of these substances in the liquid state are compiled and correlated against Le Bas molar volume in the three primary media of air, water and octanol. From these solubilities the air-water (K(AW)), octanol-water (K(OW)) and octanol-air (K(OA)) partition coefficients are deduced. Estimated solubilities in water and octanol-water partition coefficients are shown to compare favourably with more recent accurate measurements. A set of selected values is presented, with error limits, which is recommended for use in modelling and assessment studies. Some environmental implications are discussed of the large range in property values for this series. PMID- 11057576 TI - Photoinduced addition of the fungicide anilazine to cyclohexene and methyl oleate as model compounds of plant cuticle constituents. AB - Photoreactions, initiated by sunlight irradiation, between organochlorine pesticides and olefinic compounds of plant cuticles have been postulated. Concerning the formation of bound residues, which so far have not been detectable by common analytical techniques, photoaddition reactions are of main interest. In order to study the photochemical behavior of chlorinated fungicides, anilazine was irradiated in cyclohexene and methyl oleate as model compounds for olefinic plant cuticle constituents. Anilazine extensively reacted with the cis-double bond of both model compounds via radical mechanisms. In addition to a dechlorinated photoproduct several addition products were formed showing plausible reaction pathways for the formation of bound residues in plant cuticles. Photoproducts were isolated by preparative HPLC and analyzed by HPLC, MS, 1H-, and 13C-NMR. PMID- 11057577 TI - Identification of impurities in technical metalaxyl. AB - Impurities such as 2,6-dimethylaniline, N-methyl-2,6-dimethylaniline, N-(1 methoxycarbonyl-ethyl)-2,6-dimethylaniline, N-methyl-N-(1-methoxycarbonyl-ethyl) 2,6-dimethylaniline, N-methyl-N-(1-methoxyacetyl)-2,6-dimethylaniline, N-(1 methoxyacetyl)-2,6-dimethylaniline and N-ethyl-N-(methoxyacetyl)-2,6 dimethylaniline present in samples of technical metalaxyl were isolated by column chromatography and identified by nuclear magnetic resonance, mass spectroscopy and comparison with reference compounds. PMID- 11057578 TI - Assessment of metal pollution based on multivariate statistical modeling of 'hot spot' sediments from the Black Sea. AB - The paper deals with application of different statistical methods like cluster and principal components analysis (PCA), partial least squares (PLSs) modeling. These approaches are an efficient tool in achieving better understanding about the contamination of two gulf regions in Black Sea. As objects of the study, a collection of marine sediment samples from Varna and Bourgas "hot spots" gulf areas are used. In the present case the use of cluster and PCA make it possible to separate three zones of the marine environment with different levels of pollution by interpretation of the sediment analysis (Bourgas gulf, Varna gulf and lake buffer zone). Further, the extraction of four latent factors offers a specific interpretation of the possible pollution sources and separates natural from anthropogenic factors, the latter originating from contamination by chemical, oil refinery and steel-work enterprises. Finally, the PLSs modeling gives a better opportunity in predicting contaminant concentration on tracer (or tracers) element as compared to the one-dimensional approach of the baseline models. The results of the study are important not only in local aspect as they allow quick response in finding solutions and decision making but also in broader sense as a useful environmetrical methodology. PMID- 11057579 TI - Soil solutions and surface water analysis in two contrasted watersheds impacted by acid deposition, Vosges mountains, N.E. France: interpretation in terms of Al impact and nutrient imbalance. AB - Two acid watersheds in the Vosges mountains (N.E. France), one with a podzol, the other with an acid brown soil, were monitored by analysing soil solutions and two related springs for total chemistry and Al speciation. High concentrations of inorganic Al did not occur in the podzol upper horizons but were evident in the corresponding spring, together with low concentrations of Ca. In the acid brown soil area, high concentrations of inorganic Al occurred in the leaching water draining the upper soil horizons, but not in the undrained water nor in the spring, the latter exhibiting rather high Ca content. In both watersheds, needle yellowing in conifers could be observed and might be ascribed to Mg deficiency rather than to Al toxicity. PMID- 11057580 TI - Determination and identification of metabolites of the fungicides Iprodione and Procymidone in compost. AB - The main metabolites formed from Iprodione and Procymidone during the composting process have been isolated and identified by HPLC-DAD-MSD. After addition of the fungicides to the composting pile, we monitored the reaction of the two analytes and the formation of their degradation products for eight months. We verified the nature of the metabolites by comparison with those hypothesised in the literature and by comparison with the behaviour of an abiotic process in aqueous acetonitrile pH 6 and at 35 degrees C. After taking into account the different kinetic behaviours of the fungicides on degradation in compost and hydro-organic solution, breakdown pathways are proposed for biodegradation. PMID- 11057581 TI - Leaching and degradation of corn and soybean pesticides in an Oxisol of the Brazilian Cerrados. AB - Pesticide pollution of ground and surface water is of growing concern in tropical countries. The objective of this pilot study was to evaluate the leaching potential of eight pesticides in a Brazilian Oxisol. In a field experiment near Cuiaba, Mato Grosso, atrazine, chlorpyrifos, lambda-cyhalothrin, endosulfane alpha, metolachlor, monocrotofos, simazine, and trifluraline were applied onto a Typic Haplustox. Dissipation in the topsoil, mobility within the soil profile and leaching of pesticides were studied for a period of 28 days after application. The dissipation half-life of pesticides in the topsoil ranged from 0.9 to 14 d for trifluraline and metolachlor, respectively. Dissipation curves were described by exponential functions for polar pesticides (atrazine, metolachlor, monocrotofos, simazine) and bi-exponential ones for apolar substances (chlorpyrifos, lambda-cyhalothrin, endosulfane alpha, trifluraline). Atrazine, simazine and metolachlor were moderately leached beyond 15 cm soil depth, whereas all other compounds remained within the top 15 cm of the soil. In lysimeter percolates (at 35 cm soil depth), 0.8-2.0% of the applied amounts of atrazine, simazine, and metolachlor were measured within 28 days after application. Of the other compounds less than 0.03% of the applied amounts was detected in the soil water percolates. The relative contamination potentials of pesticides, according to the lysimeter study, were ranked as follows: metolachlor > atrazine = simazine >> monocrotofos > endsulfane alpha > chlorpyrifos > trifluraline > lambda cyhalothrin. This order of the pesticides was also achieved by ranking them according to their effective sorption coefficient Ke, which is the ratio of Koc to field-dissipation half-life. PMID- 11057582 TI - Heterogeneous photocatalytic decomposition of halosubstituted benzyl alcohols on semiconductor particles in aqueous media. AB - The photodegradation of 2-, 3- and 4-halosubstituted benzyl alcohols (HBAs) on semiconductive oxides (TiO2, ZnO) was studied. It was found out that the photodegradation rate increases from the fluoro to bromo derivates in the case of 2- and 4-HBAs, whereas in the case of 3-HBAs the reverse trend was observed. PMID- 11057583 TI - Anaerobic transformation of chlorophenols in methanogenic sludge unexposed to chlorophenols. AB - Transformation of all 19 chlorophenol (CP) isomers was investigated in a laboratory anaerobic methanogenic sludge that had not been exposed to synthetic chemicals. Concentration of CP was analyzed over time to calculate disappearance rate constants using first-order reaction kinetics and all possible CP degradation pathways were estimated. The rate constants ranged between 0.46 x 10( 3) and 0.161 day(-1). CPs were transformed via dechlorination. The chlorine atom at the ortho-position was the most easily dechlorinated, whereas dechlorination rate at the para-position was lowest. The overall pathways of CP transformation were much less diverse than that we previously found for contaminated sediment. The Dolfing hypothesis of microbial selection of the most thermodynamically favorable pathways was not applicable for CP transformation in this study as well as previous study performed by our group. PMID- 11057584 TI - Biodegradation of polycyclic aromatic hydrocarbons by a mixed culture. AB - We investigated the potential biodegradation of polycyclic aromatic hydrocarbons (PAHs) by an aerobic mixed culture utilizing phenanthrene as its carbon source. Following a 3-5 h post-treatment lag phase, complete degradation of 5 mg/l phenanthrene occurred within 28 h (optimal conditions determined as 30 degrees C and pH 7.0). Phenanthrene degradation was enhanced by the individual addition of yeast extract, acetate, glucose or pyruvate. Results show that the higher the phenanthrene concentration, the slower the degradation rate. While the mixed culture was also capable of efficiently degrading pyrene and acenaphthene, it failed to degrade anthracene and fluorene. In samples containing a mixture of the five PAHs, treatment with the aerobic culture increased degradation rates for fluorene and anthracene and decreased degradation rates for acenaphthene, phenanthrene and pyrene. Finally, it was observed that when nonionic surfactants were present at levels above critical micelle concentrations (CMCs), phenanthrene degradation was completely inhibited by the addition of Brij 30 and Brij 35, and delayed by the addition of Triton X100 and Triton N101. PMID- 11057585 TI - Determination of microcontaminants in sediments by on-line solid-phase extraction gas chromatography-mass spectrometry. AB - Two simple and straightforward analytical procedures for the screening of sediment samples are reported. They involve extraction with ethyl acetate or methanol and subsequent analysis by means of gas chromatography-mass spectrometry (GC-MS) using large-volume injection (LVI) or solid-phase extraction (SPE). The latter, which was originally developed for the analysis of aqueous samples, can be used without any modification. In general, 10 ml of organic solvent were added to 2 g of sediment, and the mixture was shaken and allowed to stand overnight. The methanolic extracts were then diluted in water and subjected to preconcentration and analysis using on-line SPE-GC-MS. The ethyl acetate extracts were injected directly into the GC using LVI. Both methods were used for the detection and identification of microcontaminants during a monitoring study of the river Nitra (Slovak Republic). They included polyaromatic hydrocarbons (PAHs), chlorofluorohydrocarbons, alkoxylated and alkylated phenols and benzothiazole derivatives. Semi-quantitative profiles of the contaminants were constructed and provisionally interpreted. The results indicate that SPE-GC-MS, and also LVI-GC-MS, have good potential for a rapid screening of sediment samples and the identification of microcontaminants. The analytical procedures pose no problems, and the on-line set-up is user-friendly. PMID- 11057586 TI - The nitration of pyrene adsorbed on silica particles by nitrogen dioxide. AB - Conversion of NO2, HNO2 gas, their mixture and a mixed gas of HNO2 and HNO3 on silica particles was investigated under simulated atmospheric conditions. Both HNO2 and HNO3 were detected as the products from conversion of NO2 on silica particles. However, unlike HNO3, which increased with conversion time, HNO2 underwent an increase-decrease time course due to the increased HNO3 further transformed HNO2 into NO+ on silica particles. Considering the catalytic effect of HNO3 and HNO2 on the nitration of pyrene adsorbed on silica particles by NO2, another electrophilic nitration path, analogous to the one that we previously reported, with NONO2+ and NON2O4+ as electrophiles was suggested. The two paths together gave an appropriate explanation for the catalytic effect of HNO2, HNO3 and their mixed gas on the nitration of the adsorbed pyrene by NO2. PMID- 11057587 TI - Fate of seven pesticides in an aerobic aquifer studied in column experiments. AB - The fate of selected pesticides (bentazone, isoproturon, DNOC, MCPP, dichlorprop and 2,4-D) and a metabolite (2,6-dichlorobenzamide (BAM)) was investigated under aerobic conditions in column experiments using aquifer material and low concentrations of pesticides (approximately 25 microg/l). A solute transport model accounting for kinetic sorption and degradation was used to estimate sorption and degradation parameters. Isoproturon and DNOC were significantly retarded by sorption, whereas the retardation of the phenoxy acids (MCPP, 2,4-D and dichlorprop), BAM and bentazone was very low. After lag periods of 16-33 days for the phenoxy acids and 80 days for DNOC, these pesticides were degraded quickly with 0.-order rate constants of 1.3-2.6 microg/l/day. None of the most probable degradation products were detected. PMID- 11057588 TI - Mobility of isoproturon from an alginate-bentonite controlled release formulation in layered soil. AB - The mobility of isoproturon [3-(4-isopropylphenyl)-1,1-dimethylurea] from an alginate-based controlled release (CR) formulation was investigated by using soil columns. A layered bed system simulating the typical arrangement under a plastic greenhouse, which is composed of sand, peat, amended soil and native soil was used. The CR formulation was based on sodium alginate (1.87%), isoproturon (1.19%), natural bentonite (3.28%), and water (93.66%), and was compared to technical grade isoproturon. The use of the alginate-bentonite CR formulation produced less vertical mobility of the active ingredient as compared to the technical product. There was no presence of herbicide in the leachate when the alginate-bentonite CR formulation was used. However, 0.90% of isoproturon appeared when the treatment was carried out with technical grade material. Isoproturon mobility was modelled using the programme CMLS, which showed the peat layer to retard pesticide leaching. Analysis of the soil columns showed the highest isoproturon concentration in the peat layer. PMID- 11057589 TI - Leaching properties of some degradation products of alachlor and metolachlor. AB - Once in soil, pesticides undergo degradation processes that give rise to a complex pattern of metabolites. Those presenting a significant percentage of formation, genotoxic and leaching properties may pose a threat to human health associated with the consumption of drinking water. The aim of this study is to assess the hazard potential of some metabolites that may occur in ground water. 2,6-diethylaniline, 2-chloro-2',6'-diethylacetanilide, 2-hydroxy-2',6' diethylacetanilide, metabolites of alachlor and 2-ethyl-6-methylaniline, metabolite of metolachlor, were chosen for their genotoxic properties. Under laboratory conditions, these metabolites showed DT50 = 1-5 days and Koc = 45-357. Their leaching potential, calculated according to Gustafson, is very low and, therefore, they should not be regarded as contaminants of ground waters. Aged residue leaching studies as well as preliminary studies on well waters seem to confirm these findings. PMID- 11057590 TI - Aerobic thermophilic treatment of sewage sludge contaminated with 4-nonylphenol. AB - 4-Nonylphenol (4-NP) occurs in sewage sludge as a result of the breakdown of detergents which contains nonylphenol ethoxylates (NPEs). 4-NP is of environmental concern because of its toxicity to biological systems. The present paper reports results of aerobic treatment under thermophilic conditions of sewage sludge artificially contaminated with 4-NP. Experiments were carried out using three parallel laboratory-scale batch reactors operating with blank, 50 and 100 mg/l of 4-NP concentration. For the two studied concentrations up to 66% 4-NP reduction was achieved at a specific air flow rate of 16 l/(l.h) and a thermophilic temperature of 60 degrees C, within 10 days of operation. The presence of 4-NP has minor effect on the rate of sludge oxidation and the nitrogen and phosphorous content in the sludge. PMID- 11057591 TI - Photocatalytic degradation of AZO dyes by supported TiO2 + UV in aqueous solution. AB - The photocatalytic degradation performance of photocatalysts TiO2 supported on 13 X, Na-Y, 4A zeolites with different loading content was evaluated using the photocatalytic oxidation of dyes direct fast scarlet 4BS and acid red 3B in aqueous medium. The results showed that the best reaction dosage of TiO2-zeolite catalysts is about 2 g/l and the photocatalytic kinetics follows first order for all supported catalysts. The photocatalytic activity order of the three series catalysts is 13X type >Y type >4A type. The physical state of titanium dioxide on the supports is evaluated by X-ray photoelectron spectra (XPS), powder X-ray diffraction (XRD), BET, and FTIR. PMID- 11057592 TI - An approach for assessment of water quality using semipermeable membrane devices (SPMDs) and bioindicator tests. AB - As an integral part of our continued development of water quality assessment approaches, we combined integrative sampling, instrumental analysis of widely occurring anthropogenic contaminants, and the application of a suite of bioindicator tests as a specific part of a broader survey of ecological conditions, species diversity, and habitat quality in the Santa Cruz River in Arizona, USA. Lipid-containing semipermeable membrane devices (SPMDs) were employed to sequester waterborne hydrophobic chemicals. Instrumental analysis and a suite of bioindicator tests were used to determine the presence and potential toxicological relevance of mixtures of bioavailable chemicals in two major water sources of the Santa Cruz River. The SPMDs were deployed at two sites; the effluent weir of the International Wastewater Treatment Plant (IWWTP) and the Nogales Wash. Both of these systems empty into the Santa Cruz River and the IWWTP effluent is a potential source of water for a constructed wetland complex. Analysis of the SPMD sample extracts revealed the presence of organochlorine pesticides (OCs), polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons (PAHs). The bioindicator tests demonstrated increased liver enzyme activity, perturbation of neurotransmitter systems and potential endocrine disrupting effects (vitellogenin induction) in fish exposed to the extracts. With increasing global demands on limited water resources, the approach described herein provides an assessment paradigm applicable to determining the quality of water in a broad range of aquatic systems. PMID- 11057593 TI - The photocatalytic disinfection of urban waste waters. AB - In this paper we present the results of the photocatalytic disinfection of urban waste water. Two microbial groups, total coliforms and Streptococcus faecalis, have been used as indexes to test disinfection efficiencies. Different experimental parameters have been checked, such as the effect of TiO2, solar or UV-lamp light and pH. Disinfection of water samples has been achieved employing both UV-lamp and solar light in agreement with data shown by other authors. The higher disinfection rates obtained employing an UV-lamp may be explained by the stronger incident light intensity. Nevertheless no consistent differences have been found between TiO2-photocatalysis and direct solar or UV-lamp light irradiation at natural sample pH (7.8). At pH 5 the presence of TiO2 increases the relative inactivation rate compared with the absence of the catalyst. After the photocatalytic bacterial inactivation, the later bacterial reappearance was checked for total coliforms at natural pH and pH 5, with and without TiO2. Two h after the photocatalytic treatment, CFU increment was almost nill. But 24 and 48 h later an important bacterial CFU increment was observed. This CFU increment is slower after irradiation with TiO2 at pH 5 in non-air-purged samples. PMID- 11057594 TI - Biotreatment of H2S- and NH3-containing waste gases by co-immobilized cells biofilter. AB - Gas mixture of H2S and NH3 in this study has been the focus in the research area concerning gases generated from the animal husbandry and the anaerobic wastewater lagoons used for their treatment. A specific microflora (mixture of Thiobacillus thioparus CH11 for H2S and Nitrosomonas europaea for NH3) was immobilized with Ca alginate and packed inside a glass column to decompose H2S and NH3. The biofilter packed with co-immobilized cells was continuously supplied with H2S and NH3 gas mixtures of various ratios, and the removal efficiency, removal kinetics, and pressure drop in the biofilter was monitored. The results showed that the efficiency remained above 95% regardless of the ratios of H2S and NH3 used. The NH3 concentration has little effect on H2S removal efficiency, however, both high NH3 and H2S concentrations significantly suppress the NH3 removal. Through product analysis, we found that controlling the inlet ratio of the H2S/NH3 could prevent the biofilter from acidification, and, therefore, enhance the operational stability. Conclusions from bioaerosol analysis and pressure drop in the biofilter suggest that the immobilized cell technique creates less environmental impact and improves pure culture operational stability. The criteria for the biofilter operation to meet the current H2S and NH3 emission standards were also established. To reach Taiwan's current ambient air standards of H2S and NH3 (0.1 and 1 ppm, respectively), the maximum inlet concentrations should not exceed 58 ppm for H2S and 164 ppm for NH3, and the residence time be kept at 72 s. PMID- 11057595 TI - Photooxidation mechanism of nitrogen-containing compounds at TiO2/H2O interfaces: an experimental and theoretical examination of hydrazine derivatives. AB - The photocatalytic oxidation of oxalyldihydrazide, N,N' bis(hydrazocarbonyl)hydrazide, N,N'-bis(ethoxycarbonyl)hydrazide, malonyldihydrazide, N-malonyl-bis[(N'-ethoxycarbonyl)hydrazide] was examined in aqueous TiO2 dispersions under UV illumination. The photomineralization of nitrogen and carbon atoms in the substrates into N2 gas, NH4+ (and/or NO3-) ions, and CO2 gas was determined by HPLC and GC analysis. The formation of carboxylic acid intermediates also occurred in the photooxidation process. The photocatalytic mechanism is discussed on the basis of the experimental results, and with molecular orbital (MO) simulation of frontier electron density and point charge. Substrate carbonyl groups readily adsorb on the TiO2 surface, and the bonds between carbonyl group carbon atoms and adjacent hydrazo group nitrogen atoms are cleaved predominantly in the initial photooxidation process. The hydrazo groups were photoconverted mainly into N2 gas (in mineralization yields above 70%) and partially to NH4 ions (below 10%). The formation of NO3- ions was scarcely recognized. PMID- 11057596 TI - Extraction of heavy metals from MSW incinerator fly ash using saponins. AB - An extraction process with saponins was evaluated for removing heavy metals from MSW (municipal solid waste) incinerator fly ashes. Two different fly ashes, A and B, were treated on a laboratory scale with three triterpene-glycoside type of saponins, M, Q, and T, in the pH range 4-9. The results were compared with those of the HCI and EDTA treatment. The treatment with saponins extracted 20-45% of Cr from the fly ashes. Saponins were also effective in extracting Cu from fly ash A attaining 50-60% extraction. Saponin T extracted 100% of Pb from fly ash A at pH around 4. The extraction of Zn with the saponin treatment was similar to that of the HCl treatment. Further, Cr, Cu, Pb, and Zn were fractionated by sequential extraction to investigate the effect of saponins on each fraction. Extraction behavior of other elements during the saponin treatment was also studied. The leaching test on the residues received after the saponin treatment showed that the fly ashes were successfully detoxified to meet the landfilling guideline. PMID- 11057597 TI - Degradation of short-chain alkyl- and alkanolamines by TiO2- and Pt/TiO2-assisted photocatalysis. AB - The photooxidation of C2H5NH2, (C2H5)2NH, HOC2H4NH2, (HOC2H4)2NH and (HOC2H4)3N using TiO2 and Pt/TiO2 as photocatalysts has been investigated. A laboratory set up was designed and a study on the influence of the concentration of the photocatalyst, the pH-value and the structure of the amine performed. The photocatalytic process was optimized with respect to the concentrations of the model substances during degradation. The decrease of the amine concentrations was found to be maximum at a pH of 10. The time-dependence of the formation of cationic breakdown products, such as NH3/NH4 and short-chain alkyl- and alkanolamines was studied by analyses with single column ion chromatography. The experimental data show that the photodegradation follows a Langmuir-Hinshelwood kinetic. The mineralization of the model substances also was monitored by measurements of the decrease of the TOC and of the formation of NO2 and NO3. The different mineralization efficiencies for the model substances studied are discussed with regard to their structure and adsorption behaviour on the photocatalyst. A possible breakdown mechanism involving the electrophilic attack of the hydroxyl radical is given. The applicability of the TiO2-assisted photocatalytic degradation of C2H5NH2 and (C2H5)2NH was tested at the pilot plant scale with real solar radiation. The degradation rates and products obtained were similar to those found in the laboratory experiments. PMID- 11057598 TI - Degradation of sodium 4-dodecylbenzenesulphonate photoinduced by Fe(III) in aqueous solution. AB - The Fe(III)-photoinduced degradation of 4-dodecylbenzenesulphonate (DBS) in aqueous solution was investigated. The mixing of DBS (1 mm) and Fe(III) (1 mm) solutions immediately led to the formation of a precipitate that contained DBS and monomeric Fe(OH)2+, the predominant Fe(III) species. Both species were also present in the supernatant. Irradiation of the supernatant solution resulted in a photoredox process that yielded Fe(II) and *OH radicals. The disappearance of DBS was shown to involve only attack by *OH radicals; the quantum yield of DBS disappearance is similar to the quantum yield of *OH radical formation. A wavelength effect was also observed; the rate of DBS disappearance was higher for shorter wavelength irradiation. Five photoproducts, all containing the benzene sulphonate group, were identified. *OH radicals preferentially abstract hydrogen from the carbon in the alpha position of the aromatic ring. The results show that the Fe(III)-photoinduced degradation of DBS could be used as an alternative method for polluted water treatment. PMID- 11057599 TI - Contaminated site remedial investigation and feasibility removal of chlorinated volatile organic compounds from groundwater by activated carbon fiber adsorption. AB - Groundwater contaminated by dense, non-aqueous phase liquids (DNAPLs) such as chlorinated solvents has become a serious problem in some regions of Taiwan. The sources of these contaminants are due to industrial discharges. These chlorinated volatile organic compounds (VOCs) have been proven to be carcinogenic to humans. The groundwater is used for domestic drinking water supply in some cities of Taiwan and the severely contaminated groundwater has to be treated in order to meet the requirement of drinking water standards. This study covers two areas of work. In the first part, polluted groundwater samples were collected from the contaminated site and analytical results indicated measurable concentrations of 12 representative chlorinated VOCs in water samples. The primary VOCs detected included trichloroethene (TCE), tetrachloroethene (PCE), 1,1,2-trichloroethane (1,1,2-TCA), and 1,1-dichloroethene (1,1-DCE). Second, to remove VOCs groundwater was treated using adsorption on activated carbon fiber (ACF). This involved pumping groundwater through vessels containing ACF. Most VOCs, including TCE, PCE, 1,1,2-TCA, and DCE, were readily adsorbed onto ACF and are removed from the water stream. Our study showed that the technology was able to significantly reduce chlorinated VOCs concentrations in groundwater. PMID- 11057600 TI - Photocatalytic degradation of atrazine by porphyrin and phthalocyanine complexes. AB - This study principally focused on a new kind of photochemical reaction catalyst: porphyrin and phthalocyanine complexes. In a first step, the preparation of the catalysts was optimized. A resin has been chosen to be the support of the complexes. Efficiency of catalytic activity is performed on the degradation of a pesticide: atrazine. The best atrazine degradation occurs with 4.6% of complexes versus substrate. The role of the surface has also been shown to be important. Then, their performances were demonstrated in terms of kinetics and degradation routes, compared to a classical catalyst: titanium dioxide. This study seeks to assess the efficiency of these systems both in a mercury lamp reactor and under solar irradiation which reduces energy costs. The best atrazine degradation half life found for the complexes is about 200 min with the iron phthalocyanine. These catalysts exhibit particular oxidation activities. Indeed, the degradation routes have been found different between the semi-conductor and the metallic complexes. These complexes are able to cleave the triazinic ring more efficiently than the titanium dioxide. PMID- 11057601 TI - Sorption of selected polycyclic aromatic hydrocarbons on soils in oil contaminated systems. AB - The adsorption and desorption behaviour of selected polycyclic aromatic hydrocarbons (PAHs) on different soils was investigated by static and dynamic methods. On the basis of a system including the four phases of soil, water, oil adsorbed and oil in emulsion, a model for the description of the adsorption behaviour in the presence of oil was developed. In systems without oil a similar partitioning behaviour in the batch and column experiments was observed for all PAHs. Thus the distribution coefficients can be calculated from the octanol/water coefficient of the aromatic compounds and the organic carbon content of the soils. The presence of a lipophilic phase had a significant influence on the sorption of the PAHs, usually resulting in a drastic decrease of adsorption with increasing oil content in the system. For the oil-contaminated system the modelling of the adsorption behaviour enabled a more detailed interpretation of the experimental observations as well as the calculation of the sorption behaviour for the PAHs from characteristic parameters of the components involved. PMID- 11057602 TI - Optimized combinations of abatement strategies for urban mobile sources. AB - The maximum incremental reactivity (MIR) scale was chosen as a practical index for quantifying ozone-forming impacts. The integer linear and nonlinear programming techniques were employed as the optimization method to maximize MIR and volatile organic compound (VOC) reductions, and minimize ozone's marginal cost with varied control costs. Mobile vehicles were divided into nine categories according to the demands of decision makers and the distinctive features of local circumstance in metro-Taipei. The emission factor (EF) and vehicle kilometers traveled (VKT) of each kind of vehicle were estimated by MOBILE5B model via native parameters and questionnaires. Compressed natural gas (CNG) and inspection and maintenance (I/M) were the alternative control programs for buses and touring buses; liquefied petroleum gas (LPG), I/M, methanol, electrical vehicle (EV) were for taxis and low duty gasoline vehicles. EV, methanol, and I/M were the possible control methods for two-stroke and four-stroke engine motorcycles; I/M programs for low-duty diesel trucks, heavy-duty diesel trucks, and low-duty gasoline trucks. The results include the emission ratios of specific vehicle to all vehicles, the best combination of abated measures based on different objectives, and the marginal cost for ozone and VOC with varied control costs. PMID- 11057603 TI - Quantitation of hydroxyl radical during fenton oxidation following a single addition of iron and peroxide. AB - Chemical probes were used to study the formation of hydroxyl radical in aqueous iron-hydrogen peroxide reaction. Hydroxyl radical formation rate and time dependent concentration were determined in pure water, in aqueous fulvic acid (FA) and humic acid (HA) solutions, and in natural surface waters. Indirect determinations of hydroxyl radical were made by quantitating hydroxyl radical reactions with probe compounds under controlled conditions. High probe concentrations were used to determine radical formation rates and low probe concentrations were used to determine time dependent radical concentration. Two independent probes were used for intercomparison: benzoic acid and 1-propanol. Good agreement between the two probes was observed. Natural water matrices resulted in lower radical formation rates and lower hydroxyl radical concentrations, with observed formation rate and yield in natural waters up to four times lower than in pure water. HA and FA also reduced hydroxyl radical formation under most conditions, although increased radical formation was observed with FA at certain pH values. Hydroxyl radical formation increased linearly with hydrogen peroxide concentration. PMID- 11057604 TI - Influence of modified soils on the removal of diesel fuel oil from water and the growth of oil degradation micro-organism. AB - Three soils were modified with two kinds of cationic surfactants in order to increase their sorptive capabilities for organic contaminants. Sorption of diesel fuel oil in water by these modified soils had been investigated. Modified soils can effectively sorb diesel fuel oil from water. The sorption capability of modified soils is: HDTMA-black soil > HDTMA-yellow brown soil > HDTMA-red soil > TMA-black soil > TMA-yellow brown soil > TMA-red soil. Sorption of diesel fuel oil by natural soils and HDTMA modified soils is via partition, the sorption isotherms can be expressed by Henry equation, and logK(SOM) is 2.42-2.80, logK(HDTMA) is 3.37-3.60. Sorption isotherms of TMA modified soils can be expressed by Langmuir equation, the saturation sorption capacities are 1150 (TMA black soil), 750 (TMA-yellow-brown soil), 171 mg/kg (TMA-red soil), respectively. A diesel fuel oil degradation micro-organism (Pseudomonas sp.) was isolated in the lab. To test the influence of the modified soils on the micro-organism, various growth curves of Pseudomonas in different conditions were drawn. Pseudomonas can grow very well with natural soils and TMA modified soils. The acclimation period of Pseudomonas is reduced. As to HDTMA modified soils, HDTMA loading amount is very important. When HDTMA loading amount is no higher than 0.5 CEC, the micro-organism can grow very well after a long acclimation period. PMID- 11057605 TI - Correlation between inorganic (heavy metals) and organic (PCBs and PAHs) micropollutant concentrations during sewage sludge composting processes. AB - The nature and congener composition of PCBs and PAHs present in sewage sludge composting processes was investigated. These studies included analysis of the most significant process parameters (such as pH, temperature, weight percentage variation) and in addition heavy metals whose typical composting speciation and behaviour were also considered in order to better understand organic compound time profiles. The significant correlation found between Pb, Cd, Cu and PCBs and between PAHs and Hg implies that quite a strong adsorption of PCBs onto organic matter takes place and also provides evidence for the volatilisation of PAHs. Chemical characteristics of inorganic species and organic compounds are summarised to account for the observed correlation and time trend profiles. Moreover, single congener concentrations demonstrate that the number of Cl substituents for PCBs and condensed benzene rings for PAHs determine to what extent they can be broken down for biodegradation and removed through volatilisation respectively. PMID- 11057606 TI - Volatile metabolites from mold growth on building materials and synthetic media. AB - Mold species which were isolated from damp buildings were grown on sterile building materials and some synthetic media in order to study the microbial volatile organic compounds produced. Patterns of the microbial volatile organic compounds (MVOC) were very media dependent but media which favor terpene biosynthesis may give patterns unique enough for identification of dominant indoor molds. PMID- 11057607 TI - Removal of 2,4-dichlorophenol by suspended and immobilized Bacillus insolitus. AB - In this experiment, Bacillus insolitus was isolated and selected from a mixed culture that have been acclimated to chlorophenols. Decomposition of chlorophenolic compounds will be studied using this pure culture in both suspended and immobilized form. The results are: at lower initial concentrations of 2,4-dichlorophenol (10-50 mg/l), immobilized Bacillus insolitus shows a higher removal of 2,4-dichlorophenol than Bacillus insolitus in suspended growth. When the 2,4-dichlorophenol concentration becomes higher (50-200 mg/l), both immobilized and suspended Bacillus insolitus have approximately the same efficiency for removal of 2,4-dichlorophenol. Higher concentrations of 2,4 dichlorophenol are inhibitive to the growth of either suspended or immobilized Bacillus insolitus. At lower concentrations of 2,4-dichlorophenol, immobilized mixed culture may have the same removal efficiency of 2,4-dichlorophenol as immobilized pure culture of Bacillus insolitus. But with regard to the overall 2,4-dichlorophenol removal efficiency, immobilized pure culture is considered to be superior to immobilized mixed culture. PMID- 11057608 TI - Determination of presence of fungicides by their common metabolite, 3,5-DCA, in compost. AB - This paper describes the determination of 3,5-DCA in commercial composts, a common metabolite in a class of fungicides, and dicarboximides (Vinclozolin, Chlozolinate, Iprodione, Procymidone) which are commonly used in agriculture. The extracts, obtained in acetonitrile by sonication, are analysed by HPLC/DAD without clean-up. This method has shown several advantages: reduced manipulation of samples, good recovery (80-90%) and good reproducibility (RSD% <7). The limit of detection (DL) of the analytical method has been estimated as 15 microg/kg for the common metabolite, and 35-145 microg/kg for the four fungicides in the matrices. PMID- 11057609 TI - Analysis of polycyclic aromatic hydrocarbons in sediment reference materials by microwave-assisted extraction. AB - The microwave-assisted extraction (MAE) of polycyclic aromatic hydrocarbons (PAHs) from harbor sediment reference material EC-1, marine sediment reference material HS-2 and PAH-spiked river bed soil was conducted. The extraction conditions for EC-1 were carried out at 70 degrees C and 100 degrees C under pressure in closed vessels with cyclohexane acetone (1:1), cyclohexane-water (3:1), hexane acetone (1:1), and hexane-water (3:1) for 10 min. A comparison between MAE and a 16-h Soxhlet extraction (SX) method showed that both techniques gave comparable results with certified values. MAE has advantages over the currently used Soxhlet technique due to a faster extraction time and lower quantity of solvent used. The consumption of organic solvent of the microwave method was less than one-tenth compared to Soxhlet. PMID- 11057610 TI - Antimony biomethylation by Scopulariopsis brevicaulis: characterization of intermediates and the methyl donor. AB - The filamentous fungus Scopulariopsis brevicaulis biomethylates inorganic antimony(III) compounds to trimethylstibine, that can be detected in culture headspace gases. Dimethylantimony and trimethylantimony species have been detected in the medium of these cultures, but the origin of these species was controversial. We now show that the dimethylantimony species is a true intermediate on the pathway to trimethylstibine (rather than arising from trimethylstibine oxidation or as an analytical artifact) because no dimethylantimony species are formed on trimethylstibine oxidation, as determined by using HG-GC-AAS. Furthermore, the dimethylantimony and trimethylantimony species can be separated, by using anion exchange chromatography, and so the dimethylantimony species is not an analytical artifact, formed during the hydride generation process. The antimony biomethylation mechanism was further probed by measuring incorporation of the methyl group, from 13CD3-L-methionine and CD3-D methionine, into methylantimony species and, for comparison, into methylarsenic species. The percentage incorporation of the labeled methyl group into methylarsenic and methylantimony species was not significantly different. The incorporation from 13CD3-L-methionine was 54% and 47% for antimony and arsenic, respectively. The incorporation from CD3-D-methionine was 20% and 16% for antimony and arsenic, respectively. It appears that the biomethylation of arsenic and antimony occur by very similar, perhaps identical, mechanisms. PMID- 11057611 TI - The study of TSP, PM(2.5-10) and PM2.5 during Taiwan Chi-Chi Earthquake in the traffic site of central Taiwan, Taichung. AB - Ambient particle concentration was taken on the traffic sampling site over the Chung-Chi Road over bridge (CCROB) in front of Hungkuang Institute of Technology (HKIT). The sampling time was from August 1999 to December 1999. During the sampling period, Taiwan's biggest earthquake in more than a century registered 7.3 on the Richter scale (Taiwan Chi-Chi Earthquake). Besides, there were more than 20,000 aftershocks that followed the Taiwan Chi-Chi Earthquake within three months. Thus, the PM2.5, PM(2.5-10) particle concentrations were also collected then and compared with total suspended particle (TSP) in this study. The average PM(2.5-10), PM2.5 and TSP concentrations are 24.6, 58.0 and 106 microg/m3, respectively, after the Taiwan Chi-Chi Earthquake. The average TSP concentrations before and after Taiwan Chi-Chi Earthquake were 70 and 127 microg/m3, respectively. It is clearly shown that the average concentration of TSP after Taiwan Chi-Chi Earthquake was about 1.8 times as that of TSP concentration before Taiwan Chi-Chi Earthquake in the traffic site of central Taiwan. And the ratios of PM2.5/PM(2.5-10), PM2.5/PM10 and PM2.5/TSP are 2.2%, 67.2%, 38.9%, respectively. The results also indicated about Chi-Chi fine particle concentration (PM25) and the TSP increases in the traffic site of central Taiwan after Taiwan Chi-Chi Earthquake. PMID- 11057612 TI - EOX and organochlorine compounds in fish and ringed seal samples from Lake Ladoga, Russia. AB - Information about the pollution of Lake Ladoga, the largest lake in Europe, has been controversial. Various effluents and drainage waters affect the quality of the lake water. Wastewaters have caused eutrophication of parts of Lake Ladoga, but concentrations of persistent organic pollutants in the lake's food webs are poorly understood. In this study, concentrations of some organochlorine compounds, chlorophenols (CPs), and extractable organic halogen (EOX) were determined in smelt (Osmerus eperlanus), vendace (Coregonus albula), pikeperch (Lucioperca lucioperca), whitefish (Coregonus lavaretus), and the Ladoga seal (Phoca hispida ladogensis) from the northern part of the lake. The concentrations of organochlorine compounds in fish were low. Concentrations were between 0.07 and 0.15, 0.65 and 1.0, and 0.29 and 0.48 mg/kg lipids for hexachlorobenzene, total polychlorinated biphenyl (PCB) and p,p'-DDE, respectively. The results indicated biomagnification from smelt and vendace to pikeperch and ringed seal. In ringed seals, concentrations of PCB and DDT were 12 and 29 times higher than in fish used by ringed seals as major food sources. PMID- 11057613 TI - A geometric approach to determine adsorption and desorption kinetic constants. AB - A geometric method based on Langmuir kinetics has been derived to determine adsorption and desorption kinetic constants. In the conventional procedure, either the adsorption kinetic constant (k(a)c) or desorption kinetic constant (k(d)c) is found from kinetic experiments and the other is calculated by their correlation with the equilibrium constant, i.e, k(d)c = Kcon/k(a)c, where Kcon has been known from equilibrium studies. The determined constants (Kcon, k(a)c, k(d)c), if based only on the conventional procedure, may not be accurate due to their mathematical dependence. Therefore, the objectives of this study are applying a geometric approach to directly determine Langmuir kinetic constants and describe adsorption behavior. In this approach, both adsorption kinetic constant (k(a)g) and desorption kinetic constant (k(d)g) are obtained only from data of kinetic experiments, and a geometric equilibrium constant (Kgeo) is calculated by Kgeo = k(a)g/k(d)g. The deviation between Kgeo and Kcon can prove the accuracy of k(a)g and k(d)g which were determined by this method. This approach was applicable to selenate, selenite and Mg2+ adsorption onto SiO2 regardless of whether the adsorbate formed inner- or outer-sphere complexes. However, this method showed some deviation between Kcon and Kgeo for Mn2+ adsorption because of the formation of surface Mn(II)-hydroxide clusters, which was inconsistent with the basic assumption of this method of monolayer adsorption. PMID- 11057614 TI - Prediction for biodegradability of chemicals by an empirical flowchart. AB - A method for predicting aerobic biodegradability of chemicals was developed based on empirical knowledge. A flowchart was derived from rule of thumb relationships between the biodegradability and the number of the functional groups and substructures in a certain skeletal structure of chemicals. The flowchart classified chemicals into readily biodegradable, not readily biodegradable and not predictable. It was validated by using MITI data of 177 mono benzene derivatives and 168 acyclic compounds, resulting in correct prediction at 94% and 88% levels, respectively. PMID- 11057615 TI - Levels of antifoulant Irgarol 1051 in the Conwy Marina, North Wales. AB - Irgarol 1051 (2-methylthio-4-tert-butylamino-6-cyclopropylamino-s-triazine) is an antifouling agent used in paint formulations that are applied to the hulls of ships. A survey was carried out at Conwy Marina in North Wales to determine the levels of the herbicide over a period of three months. Liquid/liquid extraction was used to concentrate the analyte for quantitative analysis using gas chromatography/mass spectrometry (GC/MS) in the selected ion monitoring (SIM) mode. The concentrations of Irgarol 1051 in Conwy marina ranged from 7 to 543 ng/l, similar to the levels found in many other marinas, estuaries and ports in England, although much lower than those in Cote d'Azur, France. The concentrations of Irgarol 1051 were not found to be influenced by salinity, pH or temperature, although there is a strong correlation between the average concentrations of Irgarol 1051 and the density of boating activity. At the levels found in the marina, it is possible that non-target photosynthetic inhibition could occur. PMID- 11057616 TI - The occurrence of trihalomethanes in the drinking water in Greece. AB - This paper summarizes and completes an investigation into the occurrence of trihalomethanes (THMs) in public water supplies in Greece. The investigation was conducted in three cities of Greece, Athens, Mytilene and Chalkida, from 1993 to 1998. Samples were collected from three treatment plants of Athens (Galatsi Treatment Plant--GTP, Menidi Treatment Plant--MTP, Kiourka Treatment Plant--KTP) and from the distribution systems of Athens, Mytilene and Chalkida. The sources for these systems are lakes, boreholes and wells. The concentrations of THMs ranged from 5 to 106 microg/l, from 4 to 27 microg/l and from 5 to 96 microg/l, in the distribution systems of Athens, Mytilene, Chalkida, respectively. The wide ranges of concentrations in three cities should be attributed to the organics concentration of raw water. In all the cases the THMs concentrations were higher during the summer and fall than in spring and winter. In GTP and in the distribution system receives water by GTP, from April 1993 to January 1995, Mytilene and Chalkida, brominated THMs dominated and existed at the highest concentration levels, whereas chloroform was the least prevalent compound, while in all the other cases the opposite was observed. In all samples the concentrations were lower than the maximum level of 100 microg/l for total trihalomethanes (TTHMs) set by European Community (EC). PMID- 11057617 TI - Formation of chloroform in spruce forest soil--results from laboratory incubation studies. AB - The release of chloroform, 1,1,1-trichloroethane, tetrachloromethane, trichloroethene and tetrachloroethene from an organic rich spruce forest soil was studied in laboratory incubation experiments by dynamic headspace analysis, thermodesorption and gas chromatography. Performance parameters are presented for the dynamic headspace system. For spruce forest soil, the results showed a significant increase in chloroform concentration in the headspace under aerobic conditions over a period of seven days, whereas the concentration of the other compounds remained fairly constant. A biogenic formation of chloroform is suggested, whereas for the other compounds anthropogenic sources are assumed. The addition of trichloroacetic acid to the soil increased the release of chloroform from the soil. It is, therefore, suggested that trichloroacetic acid also contributed to the formation of chloroform. Under the experimental conditions, the spruce forest soil released chloroform concentrations corresponding to a rate of 12 microg m(-2) day(-1). Data on chloroform production rates are presented and compared with literature results, and possible formation mechanisms for chloroform are discussed. PMID- 11057618 TI - The role of carbonate radical in limiting the persistence of sulfur-containing chemicals in sunlit natural waters. AB - Carbonate radical (*CO3-) is a selective oxidant that may be important in limiting the persistence of a number of sulfur-containing compounds in sunlit natural waters. Thioanisole, dibenzothiophene (DBT), and fenthion were selected to investigate the degradation pathway initiated by *CO3-; electron-rich sulfur compounds are particularly reactive towards the *CO3-. Using HPLC, GC, GC-MS and LC-MS for structural confirmation, the major photodegradation products of thioanisole and DBT were the corresponding sulfoxides. The sulfoxide products were further oxidized through reaction with *CO3- to the corresponding sulfone derivatives. Fenthion showed a similar pathway with appearance of fenthion sulfoxide as the major product. The proposed mechanism involves abstraction of an electron on sulfur to form a radical cation, which is then oxidized by dissolved oxygen. Each of the sulfur probes were further investigated in a sunlight simulator under varying matrix conditions. The highest rate constants occurred in the *CO3- matrix, and the lowest occurred in a matrix of dissolved organic carbon (DOC) and bicarbonate. In synthetic and natural field water, thioanisole photodegraded faster than under direct photolysis, with half-lives of 75.1 and 85.8 min, respectively. Fenthion photodegraded more rapidly than thioanisole. DBT photodegraded rapidly in a *CO3- matrix with a half-life of 24.8 min, while the half-life of direct photolysis was 350 min. Photodegradation products of each compound were also investigated. Ultimately, *CO3- was found to contribute toward the photodegradation of sulfur-containing compounds in natural waters. PMID- 11057619 TI - Effect of fuel aromatic content on PAH emission from a heavy-duty diesel engine. AB - Polycyclic aromatic hydrocarbons (PAHs) emission tests for a heavy-duty diesel engine fueled with blend base diesel fuel by adding batch fractions of poly aromatic and mono-aromatic hydrocarbons, Fluorene and Toluene, respectively, were simulated to five steady-state modes by a DC-current dynamometer with fully automatic control system. The main objective of this study is to investigate the effect of total aromatic content and poly-aromatic content in diesel fuels on PAH emission from the HDD engine exhaust under these steady-state modes. The results of this study revealed that adding 3% and 5% (fuel vol%) Fluorene in the diesel fuel increases the amount of total-PAH emission by 2.6 and 5.7 times, respectively and increases the amount of Fluorene emission by 52.9 and 152 times, respectively, than no additives. However, there was no significant variation of PAH emission by adding 10% (vol%) of Toluene. To regulate the content of poly aromatic content in diesel fuel, in contrast to the total aromatic content, will be more suitable for the management of PAH emission. PMID- 11057620 TI - Mobility of antimony in soil and its availability to plants. AB - In a historical mining area residual material has been filled on land and these locations are used today as agricultural soils or house gardens. The antimony concentrations in these soils are up to 500 mg/kg. Antimony transfer into 19 vegetable and crop species was investigated. In grain and other storage organs up to 0.09 mg Sb/kg were found, whereas maximum antimony concentrations in shoots and leaves were determined to be 0.34 mg Sb/kg and 2.2 mg Sb/kg, respectively. Despite the high antimony contamination of the soils, concentrations in the investigated plants in general corresponded to concentrations only reported for uncontaminated soils. NH4NO3 extraction of some of the soils indicated that the mobile fraction of antimony present was only 0.06-0.59%. In contrast, in leaves of spinach grown under controlled conditions in soils with a high mobile antimony content an accumulation of the element could be observed: a maximum value of 399 mg Sb/kg was detected, and a correlation between the mobile fraction in the soils and antimony in leaves was found. PMID- 11057621 TI - Monitoring, modelling and environmental exposure assessment of industrial chemicals in the aquatic environment. AB - Monitoring and laboratory data play integral roles alongside fate and exposure models in comprehensive risk assessments. The principle in the European Union Technical Guidance Documents for risk assessment is that measured data may take precedence over model results but only after they are judged to be of adequate reliability and to be representative of the particular environmental compartments to which they are applied. In practice, laboratory and field data are used to provide parameters for the models, while monitoring data are used to validate the models' predictions. Thus, comprehensive risk assessments require the integration of laboratory and monitoring data with the model predictions. However, this interplay is often overlooked. Discrepancies between the results of models and monitoring should be investigated in terms of the representativeness of both. Certainly, in the context of the EU risk assessment of existing chemicals, the specific requirements for monitoring data have not been adequately addressed. The resources required for environmental monitoring, both in terms of manpower and equipment, can be very significant. The design of monitoring programmes to optimise the use of resources and the use of models as a cost-effective alternative are increasing in importance. Generic considerations and criteria for the design of new monitoring programmes to generate representative quality data for the aquatic compartment are outlined and the criteria for the use of existing data are discussed. In particular, there is a need to improve the accessibility to data sets, to standardise the data sets, to promote communication and harmonisation of programmes and to incorporate the flexibility to change monitoring protocols to amend the chemicals under investigation in line with changing needs and priorities. PMID- 11057622 TI - Mutagenic nitrated benzo[a]pyrene derivatives in the reaction product of benzo[a]pyrene in NO2-air in the presence of O3 or under photoirradiation. AB - In order to clarify the contribution of nitrated products to the direct-mutagenic activity of products of the reactions of benzo[a]pyrene in NO2-air under various conditions, heterogeneous reactions of BaP deposited on filter in the air containing 10 ppm of NO2 have been conducted in dark or under photoirradiation. The reaction products have been analyzed by gas chromatography and mutagenicity of the products fractionated by preparative HPLC was assayed for Salmonella typhimurium strains TA98 and YG1024 in the absence of S9 mix. 3,6 dinitrobenzo[a]pyrene and 1,3-dinitrobenzo[a]pyrene, which are strong direct acting mutagens, largely contributed to the total direct-acting mutagenicity of the dark reaction products in NO2-air. On the other hand, both the dark reaction in the presence of O3 and the photoreaction in NO2-air resulted in the formation of much smaller amounts of nitrobenzo[a]pyrenes than that observed in the dark reaction in the absence of O3. These results show that the contribution of other direct-acting mutagens to the total direct-acting mutagenicity of the products in these reactions should be considered. Benzo[a]pyrene lactones were identified in a highly mutagenic fraction of the products of the dark reaction in the presence of O3 and photoreaction and a nitrobenzo[a]pyrene lactone was also identified in a highly mutagenic fraction of the dark reaction products in the presence of O3. Nitrated oxygenated benzo[a]pyrene derivatives such as nitrobenzo[a]pyrene lactone were considered to largely contribute to direct-acting mutagenicity of the products of the dark reaction in the presence of O3 and photoreaction. PMID- 11057624 TI - Desperately seeking a syndrome. PMID- 11057623 TI - Measurement of trihalomethanes in potable and recreational waters using solid phase micro extraction with gas chromatography-mass spectrometry. AB - Solid phase micro extraction (SPME) was applied to the determination of selected trihalomethanes (THMs), chloroform, bromodichloromethane, dibromochloromethane, bromoform, in potable and recreational waters. The selected samples were environmentally significant due to mandatory limits imposed by regulatory agencies. Extraction of the analytes was performed using headspace SPME (fused silica fibre with a 100 microm poly(dimethylsiloxane coating) followed by thermal desorption at 220 degrees C and GC-MS analysis. A linear working range of 10-160 microg/l was established with relative standard deviations (%RSD) within the range, 0.9-19%. Limits of detection (LOD) were 1.0-2.8 microg/l. The highest THM concentration was 61.8 microg/l which was well within the proposed European Union directive of 100 microg/l. The total THMs determined in swimming pool waters ranged from 105-134 microg/l, with chloroform accounting for 84-86% of total THM. PMID- 11057625 TI - Systems biology's multiple maths. PMID- 11057627 TI - Spain's science figures under fire. PMID- 11057626 TI - Tissue donors use their influence in deal over gene patent terms. PMID- 11057628 TI - Mbeki agrees to step back from AIDS debate. PMID- 11057629 TI - UK to make the northwest a post-genomics hotspot. PMID- 11057630 TI - NSF puts big money into complex ecology. PMID- 11057631 TI - Deep roots of Nazi science revealed. PMID- 11057633 TI - Canada plans reduction in greenhouse-gas emissions. PMID- 11057632 TI - US court slashes damages in polymerase-beta theft case. PMID- 11057634 TI - French take physics archives into the future. PMID- 11057635 TI - Anger as Princeton closes 'inspirational' museum. PMID- 11057636 TI - Leroy Hood. For my next trick... PMID- 11057637 TI - India's finest, for hire. PMID- 11057638 TI - Careers in science offer women an unusual bonus: immortality. PMID- 11057639 TI - Enigma thief stole a very special machine. PMID- 11057640 TI - Did civil reactors supply plutonium for weapons? PMID- 11057641 TI - Achievers should stay to aid Brazilian science... PMID- 11057642 TI - Yet the path is strewn with needless obstacles. PMID- 11057643 TI - If free speech costs lives that's a high price to pay. PMID- 11057644 TI - Prophets without honour? PMID- 11057645 TI - Subpoenaed in Syracuse. PMID- 11057646 TI - Identifying cosmic muck. PMID- 11057647 TI - A case of bacterial immortality? PMID- 11057648 TI - Tuning channels for blood pressure. PMID- 11057649 TI - Tracing the Earth's evolution. PMID- 11057650 TI - Mitrates on the move. PMID- 11057651 TI - A world in transition... PMID- 11057652 TI - Gene regulation. One man's food. PMID- 11057653 TI - Robert H. Abeles (1926-2000). PMID- 11057654 TI - Genital damage, kicking and early death. PMID- 11057655 TI - Is acidification still an ecological threat? PMID- 11057656 TI - A 3,000-year record of penguin populations. PMID- 11057657 TI - Glacial/interglacial variations in atmospheric carbon dioxide. AB - Twenty years ago, measurements on ice cores showed that the concentration of carbon dioxide in the atmosphere was lower during ice ages than it is today. As yet, there is no broadly accepted explanation for this difference. Current investigations focus on the ocean's 'biological pump', the sequestration of carbon in the ocean interior by the rain of organic carbon out of the surface ocean, and its effect on the burial of calcium carbonate in marine sediments. Some researchers surmise that the whole-ocean reservoir of algal nutrients was larger during glacial times, strengthening the biological pump at low latitudes, where these nutrients are currently limiting. Others propose that the biological pump was more efficient during glacial times because of more complete utilization of nutrients at high latitudes, where much of the nutrient supply currently goes unused. We present a version of the latter hypothesis that focuses on the open ocean surrounding Antarctica, involving both the biology and physics of that region. PMID- 11057658 TI - Vasoregulation by the beta1 subunit of the calcium-activated potassium channel. AB - Small arteries exhibit tone, a partially contracted state that is an important determinant of blood pressure. In arterial smooth muscle cells, intracellular calcium paradoxically controls both contraction and relaxation. The mechanisms by which calcium can differentially regulate diverse physiological responses within a single cell remain unresolved. Calcium-dependent relaxation is mediated by local calcium release from the sarcoplasmic reticulum. These 'calcium sparks' activate calcium-dependent potassium (BK) channels comprised of alpha and beta1 subunits. Here we show that targeted deletion of the gene for the beta1 subunit leads to a decrease in the calcium sensitivity of BK channels, a reduction in functional coupling of calcium sparks to BK channel activation, and increases in arterial tone and blood pressure. The beta1 subunit of the BK channel, by tuning the channel's calcium sensitivity, is a key molecular component in translating calcium signals to the central physiological function of vasoregulation. PMID- 11057659 TI - Molecular emission from single-bubble sonoluminescence. AB - Ultrasound can drive a single gas bubble in water into violent oscillation; as the bubble is compressed periodically, extremely short flashes of light (about 100 ps) are generated with clock-like regularity. This process, known as single bubble sonoluminescence, gives rise to featureless continuum emission in water (from 200 to 800 nm, with increasing intensity into the ultraviolet). In contrast, the emission of light from clouds of cavitating bubbles at higher acoustic pressures (multi-bubble sonoluminescence) is dominated by atomic and molecular excited-state emission at much lower temperatures. These observations have spurred intense effort to uncover the origin of sonoluminescence and to generalize the conditions necessary for its creation. Here we report a series of polar aprotic liquids that generate very strong single-bubble sonoluminescence, during which emission from molecular excited states is observed. Previously, single-bubble sonoluminescence from liquids other than water has proved extremely elusive. Our results give direct proof of the existence of chemical reactions and the formation of molecular excited states during single-bubble cavitation, and provide a spectroscopic link between single- and multi-bubble sonoluminescence. PMID- 11057660 TI - Strain effects and phase transitions in photonic resonator crystals. AB - Optical structures with periodic variations of the dielectric constant in one or more directions (photonic crystals) have been employed extensively for studying optical diffraction phenomena. Practical interest in such structures arises from the possibilities they offer for tailoring photon modes, and thereby the characteristics of light propagation and light-matter interactions. Photonic resonator crystals comprising two-dimensional arrays of coupled optical microcavities have been fabricated using vertical-cavity surface-emitting laser wafers. In such structures, the light propagates mostly normal to the periodic plane. Therefore, the corresponding lateral Bragg-periodicities are larger, a feature that is advantageous for device manufacture as it allows for larger lattice constants in the lateral direction. Here we investigate strain effects in a photonic resonator crystal by shifting neighbouring lattice rows of microcavities in opposite directions, thereby introducing an alternating square or quasi-hexagonal pattern of shear strain. We find that, for strain values below a critical threshold, the lasing photon mode is virtually locked to the corresponding mode supported by the unstrained photonic crystal. At the critical strain value, we observe a phase-transition-like switching between the square and quasi-hexagonal lattice modes. The tolerance of subcritical strains suggests that the resonator crystal may be useful for applications requiring high spatial coherence across the lattice, while the mode switching could potentially be exploited in free-space optical communications. PMID- 11057661 TI - Origin of ferromagnetic exchange interactions in a fullerene-organic compound. AB - Organic ferromagnets, which exhibit exchange interactions between unpaired electrons in pi-orbitals, are rare, and the origin of ferromagnetism in these compounds has so far remained unexplained. Tetrakis(dimethylamino)ethylene fullerene[60] (TDAE-C60) shows a transition to a ferromagnetic state with fully saturated s = 1/2 molecular spins at the relatively high Curie temperature (for organic materials) of 16 K (ref. 4). It has been suggested that the orientations of the C60 molecules may be important for ferromagnetism in this material, but in the absence of structural data at low temperatures there has been little progress towards understanding these microscopic interactions. Here we report the results of a comparative structural study of two different magnetic forms of TDAE-C60 crystals at low temperatures, correlating the structural properties--in particular, the intermolecular orientations--with the magnetic properties. We find that both ferromagnetism and spin-glass-like ordering are possible in this material, and depend on the orientational state of C60 molecules. This resolves the apparent contradictions posed by different macroscopic measurements, and opens the way to a microscopic understanding of pi-electron ferromagnetic exchange interactions in organic materials. PMID- 11057662 TI - Electrochemically induced annealing of stainless-steel surfaces. AB - Modification of the surface properties of metals without affecting their bulk properties is of technological interest in demanding applications where surface stability and hardness are important. When austenitic stainless steel is heavily plastically deformed by grinding or rolling, a martensitic phase transformation occurs that causes significant changes in the bulk and surface mechanical properties of the alloy. This martensitic phase can also be generated in stainless-steel surfaces by cathodic charging, as a consequence of lattice strain generated by absorbed hydrogen. Heat treatment of the steel to temperatures of several hundred degrees can result in loss of the martensitic structure, but this alters the bulk properties of the alloy. Here we show that martensitic structures in stainless steel can be removed by appropriate electrochemical treatment in aqueous solutions at much lower temperature than conventional annealing treatments. This electrochemically induced annealing process allows the hardness of cold-worked stainless steels to be maintained, while eliminating the brittle martensitic phase from the surface. Using this approach, we are able to anneal the surface and near-surface regions of specimens that contain rolling-induced martensite throughout their bulk, as well as those containing surface martensite induced by grinding. Although the origin of the electrochemical annealing process still needs further clarification, we expect that this treatment will lead to further development in enhancing the surface properties of metals. PMID- 11057663 TI - Cooler winters as a possible cause of mass extinctions at the Eocene/Oligocene boundary. AB - The Eocene/Oligocene boundary, at about 33.7 Myr ago, marks one of the largest extinctions of marine invertebrates in the Cenozoic period. For example, turnover of mollusc species in the US Gulf coastal plain was over 90% at this time. A temperature change across this boundary--from warm Eocene climates to cooler conditions in the Oligocene--has been suggested as a cause of this extinction event, but climate reconstructions have not provided support for this hypothesis. Here we report stable oxygen isotope measurements of aragonite in fish otoliths- ear stones--collected across the Eocene/Oligocene boundary. Palaeo-temperatures reconstructed from mean otolith oxygen isotope values show little change through this interval, in agreement with previous studies. From incremental microsampling of otoliths, however, we can resolve the seasonal variation in temperature, recorded as the otoliths continue to accrete new material over the life of the fish. These seasonal data suggest that winters became about 4 degrees C colder across the Eocene/Oligocene boundary. We suggest that temperature variability, rather than change in mean annual temperature, helped to cause faunal turnover during this transition. PMID- 11057664 TI - Non-chondritic distribution of the highly siderophile elements in mantle sulphides. AB - The abundances of highly siderophile (iron-loving) elements (HSEs) in the Earth's mantle provide important constraints on models of the Earth's early evolution. It has long been assumed that the relative abundances of HSEs should reflect the composition of chondritic meteorites--which are thought to represent the primordial material from which the Earth was formed. But the non-chondritic abundance ratios recently found in several types of rock derived from the Earth's mantle have been difficult to reconcile with standard models of the Earth's accretion, and have been interpreted as having arisen from the addition to the primitive mantle of either non-chondritic extraterrestrial material or differentiated material from the Earth's core. Here we report in situ laser ablation analyses of sulphides in mantle-derived rocks which show that these sulphides do not have chondritic HSE patterns, but that different generations of sulphide within single samples show extreme variability in the relative abundances of HSEs. Sulphides enclosed in silicate phases have high osmium and iridium abundances but low Pd/Ir ratios, whereas pentlandite-dominated interstitial sulphides show low osmium and iridium abundances and high Pd/Ir ratios. We interpret the silicate-enclosed sulphides as the residues of melting processes and interstitial sulphides as the crystallization products of sulphide bearing (metasomatic) fluids. We suggest that non-chondritic HSE patterns directly reflect processes occurring in the upper mantle--that is, melting and sulphide addition via metasomatism--and are not evidence for the addition of core material or of 'exotic' meteoritic components. PMID- 11057665 TI - Starch grains reveal early root crop horticulture in the Panamanian tropical forest. AB - Native American populations are known to have cultivated a large number of plants and domesticated them for their starch-rich underground organs. Suggestions that the likely source of many of these crops, the tropical forest, was an early and influential centre of plant husbandry have long been controversial because the organic remains of roots and tubers are poorly preserved in archaeological sediments from the humid tropics. Here we report the occurrence of starch grains identifiable as manioc (Manihot esculenta Crantz), yams (Dioscorea sp.) and arrowroot (Maranta arundinacea L.) on assemblages of plant milling stones from preceramic horizons at the Aguadulce Shelter, Panama, dated between 7,000 and 5,000 years before present (BP). The artefacts also contain maize starch (Zea mays L.), indicating that early horticultural systems in this region were mixtures of root and seed crops. The data provide the earliest direct evidence for root crop cultivation in the Americas, and support an ancient and independent emergence of plant domestication in the lowland Neotropical forest. PMID- 11057666 TI - Isolation of a 250 million-year-old halotolerant bacterium from a primary salt crystal. AB - Bacteria have been found associated with a variety of ancient samples, however few studies are generally accepted due to questions about sample quality and contamination. When Cano and Borucki isolated a strain of Bacillus sphaericus from an extinct bee trapped in 25-30 million-year-old amber, careful sample selection and stringent sterilization techniques were the keys to acceptance. Here we report the isolation and growth of a previously unrecognized spore forming bacterium (Bacillus species, designated 2-9-3) from a brine inclusion within a 250 million-year-old salt crystal from the Permian Salado Formation. Complete gene sequences of the 16S ribosomal DNA show that the organism is part of the lineage of Bacillus marismortui and Virgibacillus pantothenticus. Delicate crystal structures and sedimentary features indicate the salt has not recrystallized since formation. Samples were rejected if brine inclusions showed physical signs of possible contamination. Surfaces of salt crystal samples were sterilized with strong alkali and acid before extracting brines from inclusions. Sterilization procedures reduce the probability of contamination to less than 1 in 10(9). PMID- 11057667 TI - The proteins of linked genes evolve at similar rates. AB - Much more variation in the rate of protein evolution occurs than is expected by chance. But why some proteins evolve rapidly but others slowly is poorly resolved. It was proposed, for example, that essential genes might evolve slower than dispensable ones, but this is not the case; and despite earlier claims, rates of evolution do not correlate with amino-acid composition. A few patterns have been found: proteins involved in antagonistic co-evolution (for example, immune genes, parasite antigens and reproductive conflict genes) tend to be rapidly evolving, and there is a correlation between the rate of protein evolution and the mutation rate of the gene. Here we report a new highly statistically significant predictor of a protein's rate of evolution, and show that linked genes have similar rates of protein evolution. There is also a weaker similarity of rates of silent site evolution (see ref. 13), which appears to be, in part, a consequence of the similarity in rates of protein evolution. The similarity in rates of protein evolution is not a consequence of underlying mutational patterns. A pronounced negative correlation between the rate of protein evolution and a covariant of the recombination rate indicates that rates of protein evolution possibly reflect, in part, the local strength of stabilizing selection. PMID- 11057668 TI - Metapopulation dynamics of bubonic plague. AB - Bubonic plague is widely regarded as a disease of mainly historical importance; however, with increasing reports of incidence and the discovery of antibiotic resistant strains of the plague bacterium Yersinia pestis, it is re-emerging as a significant health concerns. Here we bypass the conventional human-disease models, and propose that bubonic plague is driven by the dynamics of the disease in the rat population. Using a stochastic, spatial metapopulation model, we show that bubonic plague can persist in relatively small rodent populations from which occasional human epidemics arise, without the need for external imports. This explains why historically the plague persisted despite long disease-free periods, and how the disease re-occurred in cities with tight quarantine control. In a contemporary setting, we show that human vaccination cannot eradicate the plague, and that culling of rats may prevent or exacerbate human epidemics, depending on the timing of the cull. The existence of plague reservoirs in wild rodent populations has important public-health implications for the transmission to urban rats and the subsequent risk of human outbreaks. PMID- 11057669 TI - Involuntary orienting to sound improves visual perception. AB - To perceive real-world objects and events, we need to integrate several stimulus features belonging to different sensory modalities. Although the neural mechanisms and behavioural consequences of intersensory integration have been extensively studied, the processes that enable us to pay attention to multimodal objects are still poorly understood. An important question is whether a stimulus in one sensory modality automatically attracts attention to spatially coincident stimuli that appear subsequently in other modalities, thereby enhancing their perceptual salience. The occurrence of an irrelevant sound does facilitate motor responses to a subsequent light appearing nearby. However, because participants in previous studies made speeded responses rather than psychophysical judgements, it remains unclear whether involuntary auditory attention actually affects the perceptibility of visual stimuli as opposed to postperceptual decision and response processes. Here we provide psychophysical evidence that a sudden sound improves the detectability of a subsequent flash appearing at the same location. These data show that the involuntary orienting of attention to sound enhances early perceptual processing of visual stimuli. PMID- 11057670 TI - Ghrelin induces adiposity in rodents. AB - The discovery of the peptide hormone ghrelin, an endogenous ligand for the growth hormone secretagogue (GHS) receptor, yielded the surprising result that the principal site of ghrelin synthesis is the stomach and not the hypothalamus. Although ghrelin is likely to regulate pituitary growth hormone (GH) secretion along with GH-releasing hormone and somatostatin, GHS receptors have also been identified on hypothalamic neurons and in the brainstem. Apart from potential paracrine effects, ghrelin may thus offer an endocrine link between stomach, hypothalamus and pituitary, suggesting an involvement in regulation of energy balance. Here we show that peripheral daily administration of ghrelin caused weight gain by reducing fat utilization in mice and rats. Intracerebroventricular administration of ghrelin generated a dose-dependent increase in food intake and body weight. Rat serum ghrelin concentrations were increased by fasting and were reduced by re-feeding or oral glucose administration, but not by water ingestion. We propose that ghrelin, in addition to its role in regulating GH secretion, signals the hypothalamus when an increase in metabolic efficiency is necessary. PMID- 11057671 TI - Repressor activity of Headless/Tcf3 is essential for vertebrate head formation. AB - The vertebrate organizer can induce a complete body axis when transplanted to the ventral side of a host embryo by virtue of its distinct head and trunk inducing properties. Wingless/Wnt antagonists secreted by the organizer have been identified as head inducers. Their ectopic expression can promote head formation, whereas ectopic activation of Wnt signalling during early gastrulation blocks head formation. These observations suggest that the ability of head inducers to inhibit Wnt signalling during formation of anterior structures is what distinguishes them from trunk inducers that permit the operation of posteriorizing Wnt signals. Here we describe the zebrafish headless (hdl) mutant and show that its severe head defects are due to a mutation in T-cell factor-3 (Tcf3), a member of the Tcf/Lef family. Loss of Tcf3 function in the hdl mutant reveals that hdl represses Wnt target genes. We provide genetic evidence that a component of the Wnt signalling pathway is essential in vertebrate head formation and patterning. PMID- 11057672 TI - Development of Th1-type immune responses requires the type I cytokine receptor TCCR. AB - On antigen challenge, T-helper cells differentiate into two functionally distinct subsets, Th1 and Th2, characterized by the different effector cytokines that they secrete. Th1 cells produce interleukin (IL)-2, interferon-gamma (IFN-gamma) and lymphotoxin-beta, which mediate pro-inflammatory functions critical for the development of cell-mediated immune responses, whereas Th2 cells secrete cytokines such as IL-4, IL-5 and IL-10 that enhance humoral immunity. This process of T-helper cell differentiation is tightly regulated by cytokines. Here we report a new member of the type I cytokine receptor family, designated T-cell cytokine receptor (TCCR). When challenged in vivo with protein antigen, TCCR deficient mice had impaired Th1 response as measured by IFN-gamma production. TCCR-deficient mice also had increased susceptibility to infection with an intracellular pathogen, Listeria monocytogenes. In addition, levels of antigen specific immunoglobulin-gamma2a, which are dependent on Th1 cells, were markedly reduced in these mice. Our results demonstrate the existence of a new cytokine receptor involved in regulating the adaptive immune response and critical to the generation of a Th1 response. PMID- 11057673 TI - The nuclear receptor CAR mediates specific xenobiotic induction of drug metabolism. AB - Organisms encounter a wide range of foreign compounds--or 'xenobiotics'--with potentially harmful consequences. The cytochrome P450 (CYP) enzymes metabolize xenobiotics and thus are a primary defence against these compounds. Increased expression of specific CYP genes in response to particular xenobiotics is a central component of this defence, although such induction can also increase production of toxic metabolites. Here we show that the nuclear receptor CAR mediates the response evoked by a class of xenobiotics known as the 'phenobarbital-like inducers'. The strong activation of Cyp2b10 gene expression by phenobarbital, or by the more potent TCPOBOP, is absent in mice lacking the CAR gene. These animals also show decreased metabolism of the classic CYP substrate zoxazolamine and a complete loss of the liver hypertrophic and hyperplastic responses to these inducers. Cocaine causes acute hepatotoxicity in wild-type mice previously exposed to phenobarbital-like inducers and this toxicity is also absent in the CAR-deficient animals. Thus, loss of CAR function alters sensitivity to toxins, increasing or decreasing it depending on the compound. Modulation of CAR activity in humans may significantly affect metabolism of drugs and other xenobiotics. PMID- 11057674 TI - Structure of a serpin-protease complex shows inhibition by deformation. AB - The serpins have evolved to be the predominant family of serine-protease inhibitors in man. Their unique mechanism of inhibition involves a profound change in conformation, although the nature and significance of this change has been controversial. Here we report the crystallographic structure of a typical serpin-protease complex and show the mechanism of inhibition. The conformational change is initiated by reaction of the active serine of the protease with the reactive centre of the serpin. This cleaves the reactive centre, which then moves 71 A to the opposite pole of the serpin, taking the tethered protease with it. The tight linkage of the two molecules and resulting overlap of their structures does not affect the hyperstable serpin, but causes a surprising 37% loss of structure in the protease. This is induced by the plucking of the serine from its active site, together with breakage of interactions formed during zymogen activation. The disruption of the catalytic site prevents the release of the protease from the complex, and the structural disorder allows its proteolytic destruction. It is this ability of the conformational mechanism to crush as well as inhibit proteases that provides the serpins with their selective advantage. PMID- 11057675 TI - Toxicity of fenvalerate to caddisfly larvae: chronic effects of 1- vs 10-h pulse exposure with constant doses. AB - Episodic pollution events such as runoff or spraydrift can lead to a short-term (few hours) contamination of aquatic ecosystems with pesticides. So far, different short-term exposures with respect to long-term effects have not been studied. In the present study, caddisfly larvae, typical for agricultural streams (Limnephilus lunatus Curtis, 2nd and 3rd instar) were exposed for 1- vs 10-h to three different equivalent doses (microg h) of fenvalerate. After transfer into an artificial stream microcosm with pesticide-free water, chronic effects were observed over 240 days. Comparison of 1- and 10-h exposure revealed that 1-h contamination leads to stronger effects. The differences were significant for the sublethal endpoints emergence pattern and dry weight of adults (ANOVA, Fisher's PLSD; P < 0.05). In terms of exposure dose, the difference between 1- and 10-h exposure equals a factor of 6 as a mean of all endpoints studied. The following significant effect levels for the 1-h exposure were obtained for the different endpoints investigated: reduced emergence success and production at 0.1 microg l( 1), temporal pattern of emergence at 0.001 microg l(-1), dry weight of adults at 0.01 microg l(-1). PMID- 11057676 TI - The cytotoxic effect of wastewater from the phosphoric gypsum depot on common oak (Quercus robur L.) and shallot (Allium cepa var. ascalonicum). AB - The effect of wastewater from a phosphoric gypsum depot on common oak, Quercus robur L., at cytogenetical level was studied. Allium-test was used as a control. The treatment of common oak seedlings with wastewater under laboratory conditions caused mitodepressive effect. Chromosome aberrations and mitotic irregularities were found. Cytogenetic analysis of common oak seedlings grown from acorns collected near the depot did not show changes in mitotic activity in comparison to control but the number of aberrations was higher than in control. In comparison to Alliumtest common oak was found to be more tolerant to wastewater from the phosphoric gypsum depot. PMID- 11057677 TI - A screening level examination of the potential risks of acetone in aquatic and terrestrial environments using multi-media modeling. AB - Approximately half of the approximately 40 million tonnes per annum (t/a) of acetone released worldwide arises via natural processes. The remaining releases of acetone are the focus of this assessment and arise either as a photo degradation by-product of other organic compounds (approximately 20 million t/a) or by entering the environment from manufacturing and end uses (59,000 t/a). Multi-media modeling was used to estimate regional concentrations of acetone in air, water, soil and sediment that may occur based on these anthropogenic releases to the environment. US toxics release inventory data were used to calculate local surface water concentrations. The distributions of all regional and local concentrations in all media were below applicable predicted no effect concentrations (PNECs). Calculated regional and local concentrations of acetone, originating from all anthropogenic sources, appear unlikely to cause adverse risks to the environment. PMID- 11057678 TI - Calcium chloride and calcium bromide aqueous solutions of technical and analytical grade in Lemna bioassay. AB - Saturated water solutions of calcium chloride, calcium bromide and their 1:1 mixture are commonly used as "high density brines" for pressure control in oil wells. To compare the effect of these chemicals of technical grade with the effect of the chemicals of analytical grade the Lemna test was used. The multiplication rate, fresh weight, dry to fresh weight ratio, area covered by plants and chlorophyll content were measured as toxicity parameters. The concentrations of tested chemicals were 0.025, 0.05. 0.075 and 0.1 mol dm(-3). Generally, the chemicals of both technical and analytical grade in concentrations of 0.025 mol dm(-3) stimulated the Lemna minor growth, while tested chemicals in concentrations of 0.05 mol dm(-3) did not affect the growth significantly. The exceptions were results obtained by measuring fresh weight. Most of tested chemicals in concentrations of 0.075 mol dm(-3) and all chemicals in concentrations of 0.1 mol dm(-3) reduced the growth. No major differences between effects of tested chemicals of technical and analytical grade on plant growth were observed, except that tested chemicals of analytical grade in concentrations of 0.1 mol dm(-3) increased dry to fresh weight ratio much stronger than chemicals of technical grade. All tested chemicals in all concentrations increased chlorophyll content. After treatment with chemicals of analytical grade much higher increase of chlorophyll a concentration in comparison to increase of chlorophyll b was noticed, while chemicals of technical grade caused more prominent increase of chlorophyll b. PMID- 11057679 TI - A comparison of methods for measuring acute toxicity to Hydra vulgaris. AB - The freshwater coelenterate Hydra vulgaris (Pallas) is a common component of freshwater ecosystems and is sensitive to a range of pollutants. It can be cultured easily in the laboratory to provide large numbers of test animals and can be used in simple, cost effective bioassays of both acute, sub-acute and chronic toxicity. This study, using the heavy metals: copper, cadmium and zinc, aimed to compare two methods of assessing acute toxicity. The first method was based upon a conventional determination of LC50s while the second involved an evaluation, using a scoring procedure, of progressive changes in structure and proved to have a number of advantages compared with a simple LC50. These include the potential to help explain mechanisms of toxicity and to examine the ability of animals to recover from pollutant-induced damage. PMID- 11057680 TI - Use of a lux-marked rhizobacterium as a biosensor to assess changes in rhizosphere C flow due to pollutant stress. AB - The flow of carbon from plant roots into soil supports a range of microbial processes and is therefore critical to ecosystem function and health. Pollution induced stress, which influences rhizosphere C flow is of considerable potential importance, and therefore needs to be evaluated. This paper reports on a method, based on reporter gene technology, for quantifying pollutant effects on rhizosphere C flow. The method uses the lux-marked rhizobacterium Pseudomonas fluorescens, where bioluminescence output of this biosensor is directly correlated with the metabolic activity and reports on C flow in root exudate. Plantago lanceolata was treated with paraquat (representing a model pollutant stress) in a simple microcosm system. The lux-biosensor response correlated closely with C concentrations in the exudate and demonstrated that the pollutant stress increased the C flow from the plantago roots, 24 h after application of the herbicide. The lux-reporter system therefore potentially offers a technique for use in assessing the impact of pollutant stress on rhizosphere C flow through the soil microbial biomass. PMID- 11057681 TI - Environmental contaminants and biochemical response in eel exposed to Po river water. AB - Groups of eels (Anguilla anguilla) were exposed to Po river water under control conditions at "la Casella" Fluvial Hydrobiology Station for 30-day periods between November and May. At the end of the exposure period, fish were sacrificed and cytochrome P4501A was evaluated in liver microsomes using both catalytic (EROD) and semiquantitative immunodetection (ELISA) assays. At the same time, water samples were taken for chemical analyses of PCBs, PAHs and pesticides. Eel was chosen as bio-indicator on the bases of experiments on cytochrome P450 expression and activity under basal or induced conditions and in consideration of the ecological and economic relevance and ease of handling. Low levels of cytochrome P450 during Winter were followed by a step increase during Spring, associated with a substantial concentration increase of agrochemicals. A good correlation was found between measurements of cytochrome P4501A by EROD and ELISA in dose-effect experiments, however, ELISA showed a higher sensitivity. Immunochemical techniques may be used in addition to enzyme activity measurements both to detect lower levels of cytochrome P450 induction (where they proved more sensitive) and as quality control. These results suggest that measurement of cytochrome P4501A under controlled conditions, using eel as bio-indicator, can be a useful tool in monitoring Po river ecosystems. PMID- 11057682 TI - Fragment constant method for prediction of fish bioconcentration factors of non polar chemicals. AB - A fragment constant method for prediction of fish bioconcentration factor (BCF) was established based on experimental BCF values for 80 non-polar chemicals from nine classes. The model was evaluated using coefficients of determination and mean residuals, which are 0.995 and 0.1836, respectively. Jackknife tests were applied to examine the robustness of the prediction model on a class-by-class basis. PMID- 11057683 TI - Assessment of environmental endocrine disruptors in bald eagles of the Great Lakes. AB - Environmental endocrine disruption in wildlife has primarily focused on estrogenic/androgenic end points and their antagonists. We describe here the work that has occurred within the Great Lakes of North America that has used the bald eagle (Haliaeetus leucocephalus) as a sentinel species of the effects of environmental toxicants, including endocrine disruption. Our data suggests that population level effects of hormone disrupting chemicals, not necessarily estrogen/androgen mimics and their antagonists, have been associated with reproductive and teratogenic effects observed in the bald eagle population within the Great Lakes Basin. Additional laboratory and field studies are necessary to further clarify the role of environmental endocrine disruptors on reproduction in avian populations. The use of sea eagles (Haliaeetus spp.) as biosentinels of pollution in other regions of the world is also discussed. PMID- 11057684 TI - Hematology and serum chemistries of nestling bald eagles (Haliaeetus leucocephalus) in the lower peninsula of MI, USA. AB - Hematology constituents and serum biochemistries were determined in blood collected from 55 nestling bald eagles (Haliaeetus leucocephalus) from nest sites within the lower peninsula of Michigan in 1992. Hematological values were comparable to published ranges for birds for all but eosinophils, which were greater than normal. Serum chemistry values were similar to those of other birds for all but six parameters, uric acid, cholesterol, alkaline phosphatase, total protein, globulin, and urea nitrogen, which were greater and glucose which was less. Samples of blood collected from wild bald eagles can be used for hematologic parameters and serum chemistry. It is important for other studies of endangered species to obtain baseline data from healthy, wild animals in their natural environment, and for comparison of animals living in environments of greater exposure to those living in areas of lesser exposure to xenobiotics. We caution that arrangements for rapid analysis be done in advance of sample collection. PMID- 11057685 TI - Exposure of Chironomus riparius larvae (diptera) to lead, mercury and beta sitosterol: effects on mouthpart deformation and moulting. AB - Mouthpart deformation in chironomid larvae is induced by exposure to chemical contaminants and is becoming an established bio-indicator in sediment assessment programmes. However, concentration-response relationships with causal agents have only been established occasionally and with varying success. In this laboratory study, instar II and III larvae were exposed to sub-lethal concentrations of lead, mercury and beta-sitosterol. A significant deformation response was induced in the pecten with lead and mercury. Deformation frequencies of the mentum after metal exposures were not significantly different from the control. Moulting was retarded by both metals and was well correlated with mouthpart deformation. The beta-sitosterol is an endocrine disruptor, which was used to test the hypothetical cause-effect relation between disruption of ecdyson functioning and chironomid deformation. In the present study, exposure to sublethal concentrations of beta-sitosterol did not result in any effect on deformation or moulting. As such, the proposed hypothesis of endocrine disruptors as primary causal agents of chironomid deformation could not be substantiated. Acetone, which was used as a solvent to apply beta-sitosterol caused a significant increase of mentum deformation. The ground filtration paper used as substrate seemed to induce deformities as well. Substrate contamination, acetone and (especially) inbreeding were most probably responsible for the high deformation frequencies in the control conditions. PMID- 11057686 TI - Tissue, sex and age specific accumulation of heavy metals (Zn, Cu, Pb, Cd) by populations of the mole (Talpa europaea L.) in a central urban area. AB - Heavy metal accumulation was studied in urban populations of moles, Talpa europaea L. (Insectivora) at three sites along the "Donaukanal" in Vienna. Kidneys, liver, lung, gonads, spleen, pancreas, femur, stomach + content, heart and skin were analysed for Zn, Cu, Pb and Cd contents. Female moles showed higher Zn concentrations in spleen, gonads and skin, higher Cd concentrations in femur and stomach and lower Cu content in kidneys compared to males. Age classes were determined using teeth section. A significant increase in Cd concentration with age was found in kidneys, liver, heart, lung, skin and femur, with the highest increase for kidney samples. Pb accumulated in bone tissue (femur) with increasing age. Although significant site differences in heavy metal concentrations of soil and vegetation samples existed, these differences were only partly reflected by tissue contents. PMID- 11057687 TI - Effects of halone 1301 on Lepidium sativum, Petunia hybrida and Phaseolus vulgaris. AB - Halone 1301 belongs to a group of widely used fire repellants. Although banned in several countries, the production has still not been discontinued, and thus hazards due to use or spill can be expected. The study reports on effects of the halone 1301 on three plant species frequently used for bioindication studies: Lepidium sativum (mouse-ear cress), Phaseolus vulgaris (bush bean) and Petunia hybrida. Plants were exposed to 1 ppbv of the gas in ambient air under controlled conditions for 18 days (L. sativum), and 45 days (P. vulgaris, P. hybrida), respectively. None of the plants showed visible stress symptoms. Chlorophylls in cress and petunia were unaffected whereas in beans significant changes of the photosynthetic pigments were observed. Photosynthesis and gas exchange of bean plants were monitored during the experiment, and a lowering of transpiration was noticed. In all investigated plants, protein contents declined significantly, but despite this reduction, activity of the glutathione S-transferases (GST) increased strongly in bean and petunia. The significance of this reaction as detoxification step is discussed. PMID- 11057688 TI - Toxicity and detoxification of Swedish detergents and softener products. AB - Detergents and softeners are used in large quantities and some of their ingredients are highly toxic to aquatic organisms. In the present study the acute toxicity to Daphnia magna was determined for 26 detergents and five softener Swedish products. Only one of the detergents had a 48-h EC50 > 100 mg/l. The 48-h EC50 for the other 25 detergents ranged from 4 to 85 mg/l. The 48-h EC50 for the five softeners ranged from 15 to 166 mg/l. Detoxification tests, with and without inoculum of sewage organisms, showed that all tested products were detoxified to some extent after 16 days and that the rate of detoxification was considerably higher with addition of sewage organisms. Toxicity to D. magna of the detergents and softeners, and the biotic detoxification rate was correlated with the concentration of surfactants used in formulating the products (more surfactants increased toxicity and a slower rate of detoxification). These results emphasize the importance of biological purification of domestic wastewater containing detergents and a suggested development of less toxic and more easily degradable surfactants. PMID- 11057689 TI - Ecotoxicological soil evaluation by FETAX. AB - The frog embryo teratogenesis assay-Xenopus (FETAX) is a powerful and flexible bioassay that makes use of the embryos of the anuran Xenopus laevis. FETAX satisfies the requirements of low cost, reliability and reproducibility and, thanks to its three endpoints (i.e., mortality, teratogenicity and growth inhibition) can detect the xenobiotics that affect embryonic development. In this paper, we have used FETAX to evaluate samples of soils collected in an oil contaminated area. Embryos were exposed directly to the soil to be tested. Particular attention was devoted to provide a statistical procedure for analysing mortality and malformation data as well as growth retardation. PMID- 11057690 TI - Ecotoxicological evaluation of pig slurry. AB - Swine sewage could be source of nutrients and pollutants. This work estimates the environmental risk in nine samples from different farm treatment systems based on the evaluation of their effects in Daphnia magna acute test, and on the assessment of Cu, Zn and ammonia as main contributors. NH3 and Cu were responsible for LC50 results (1-5% of dilution). Organic compounds were quantified through several extraction methods (SPMDs, SPE and solvent extraction). A more exhaustive extraction was performed in an additional sample, which showed indole and phenol recoveries much higher than the previous ones. This method also includes PCBs (430 ppb) and fatty acids (approximately =150 ppm) quantification. PMID- 11057691 TI - Yeasts as a model for assessing the toxicity of the fungicides Penconazol, Cymoxanil and Dichlofluanid. AB - In the present work the sensitivity of yeast strains of Kluyveromyces marxianus, Pichia anomala, Candida utilis, Schizosaccharomyces pombe and Saccharomyces cerevisiae, to the fungicides cymoxanil, penconazol, and dichlofluanid, was evaluated. Dichlofluanid induced the most negative effects, whereas penconazol in general was not very toxic. Overall, our results show that the parameters IC50 for specific respiration rates of C. utilis and S. cerevisiae and C(D) for cell viability of S. cerevisiae can be applied to quantify the toxicity level of the above compounds in yeast. Hence, could be explored as an alternative or at least as a complementary test in toxicity studies and, therefore, its potential for inclusion in a tier testing toxicity test battery merits further research. PMID- 11057692 TI - Toxicology of benzyl alcohols: a QSAR analysis. AB - There is an evidence that benzyl alcohols may exhibit toxicity via a radical mechanism. To test this possibility, we studied the toxicity of para substituted benzyl alcohols on rapidly dividing cancer cells (L1210 leukemia). This system has previously found utility in studying the apparent radical toxicity of a variety of phenols. However, no evidence could be found for an electronic effect and the cellular toxicity was associated primarily with hydrophobicity. Comparison of this quantitative structure-activity relationships (QSAR) with others for the reactions of benzyl alcohols in diverse systems provides insight into mechanisms of action. A QSAR for the interaction of benzyl alcohols with protozoa yields an equation that is dependent on both hydrophobicity and acidity of the OH group versus a mixture of bacteria and fungi, the critical dependence on hydrophobicity prevails with a small dependence on a resonance-stabilized, radical mediated electronic effect. The chloramphenicols provide an instructive example, where the radical mediated electronic effect overshadows the hydrophobic contribution to bacterial toxicity. These various QSAR for benzyl alcohols indicate that mechanisms of growth inhibition in vitro vary depending on cell/organism type, the strength of the bond and lability of the hydrogen, and the strength of the initiating radical reagent. PMID- 11057693 TI - The effect of cadmium on oogenesis in Xenopus laevis. AB - Reproductive toxicity studies have historically centered on post-fertilization events. A thorough assessment of reproductive hazards to an organism should include all aspects of its life cycle. Cadmium is a teratogenic and carcinogenic heavy metal that occurs naturally in the environment but is also released anthropogenically. The effect of cadmium administration on oocyte development in Xenopus laevis was studied. Adult female Xenopus were injected in the dorsal lymph sac with cadmium chloride (CdCl2) at doses of 0.5, 0.75, 1.0, 3.0 or 5.0 mg/kg every other day for 21 days. Significant adverse effects of Cd on oocyte development were observed. The percentage of oocytes at all stages of oogenesis was decreased while the population of atretic oocytes increased dramatically (P < 0.0001). Numerous oocytes exhibited a speckled or mottled appearance and the incidence of completely atretic oocyte follicles increased. The observations indicate that Cd has the potential to significantly disrupt oogenesis and that examination of developing gametes may be a useful parameter for assessing the influence of environmental contaminants on reproductive capacity. PMID- 11057694 TI - Evaluation of surrogate measures of cadmium, lead, and zinc bioavailability to Eisenia fetida. AB - We evaluated weak-electrolyte (0.1 M Ca(NO3)2) soil extractions and ion-exchange membranes coated with a metal chelator as measures of Cd, Pb, and Zn bioavailability in spiked artificial soil by comparing their metal availability estimates to acute lethal toxicity in the earthworm Eisenia fetida. Ca(NO3)2 extractions were precisely related to toxicity in all toxicity tests, and enabled the development of time-independent LC50S (incipient lethal-levels, ILLs) calculated using exposure levels based on extraction data. ILLs with 95% CIs for the Cd, Pb, and Zn toxicity tests were 9.8 (9.4-10.3), 1.16 (1.11-1.22), and 6.33 (6.18-6.49) Ca(NO3)2-extractable mmol metal/kg soil, respectively. Mixture toxicity of Cd, Pb, and Zn, assessed using the toxic unit (TU) approach, was 1.35 TU, suggesting additivity. Chelating ion-exchange membrane uptake was variable, and not well related to toxicity. Weak-electrolyte extractions show promise as precise, inexpensive surrogate measures of Cd, Pb, and Zn bioavailability in soil. PMID- 11057695 TI - The acute and chronic toxicity of lanthanum to Daphnia carinata. AB - The rare earth elements (REEs) are increasingly being used as trace supplements in agriculture. This study measured the acute and chronic toxicity of one REE, lanthanum (La), to Daphnia carinata. The 48-h EC50 of La to Daphnia was measured in three media of differing composition and hardness. Lanthanum was most toxic to Daphnia in soft tap water (TW) with an acute 48-h EC50 of 43 microg/l compared with 1180 microg/l in ASTM hard water (ASTM). In the third daphnid growth medium (DW), based on diluted sea water, the acute 48-h EC50 was 49 microg La/l, however, there was significant precipitation of La in this media. The chronic toxicity of La to Daphnia was measured in the DW and ASTM media. Nominal exposure concentrations were 100, 200, 400, 600, 800, and 1000 microg La/l. Mortality was a more sensitive endpoint than growth or reproduction in both chronic experiments. Very little La was detected in either media after 24 h and the measured concentrations below were estimated by logarithmic mean of nominal and measured values. There was 100% mortality at concentrations > or = 80 microg La/l (400 microg/l nominal) by day six of the experiment using DW media, but no effect on survival growth or reproduction at lower concentrations. In the ASTM media, La caused significant mortality to Daphnia at concentrations > or = 39 microg/l (200 microg/l nominal), however, at least one animal survived to the end of the study at each of the tested concentrations. There was no effect of La on growth of surviving daphnids at concentrations < or = 57 microg/l (400 g/l), however, second brood clutch sizes were significantly increased at 30, 39, and 57 microg/l (100, 200, 400 g/l nominal) compared with controls. Lanthanum also caused a delayed maturation in Daphnia. PMID- 11057696 TI - Estimation of bioconcentration factors of nonionic organic compounds in fish by molecular connectivity indices and polarity correction factors. AB - A bioconcentration factor (BCF) estimation model for a wide range of nonionic organic compounds was developed on the basis of molecular connectivity indices and polarity correction factors. The nonlinear topological modeling using polarity correction factors resulted in the best BCF estimation quality for all of the 239 compounds studied, with a mean absolute estimation error of 0.478 log units. Residual analysis indicated that the estimation errors came from many sources including BCF measurement, test species, and selection of descriptors. Statistical robustness of the developed model was validated by modified jackknifed tests where random deletion of a set of compounds and specific deletion of a class of compounds were both performed. Comparison between the MCI based (molecular connectivity indices) model and a Kow-based (octanol/water partition coefficient) model revealed that the BCF estimation based on topological parameters was as good as that achieved by Kow. PMID- 11057697 TI - Hepatotoxicity of monobromobenzene and hexabromobenzene: effects of repeated dosage in rats. AB - The aim of the study was to determine whether monobromobenzene (BB) and hexabromobenzene (HBB) administered repeatedly (for 28 days) to female rats resulted in disturbances of heme synthesis. 5-Aminolevulinate dehydratase (ALA-D) and 5-aminolevulinate synthase (ALA-S) activities were slightly changed and the concentration of glutathione increased. The excretion of 5-aminolevulinic acid (ALA-U) in urine after all doses of BB and HBB increased already in the first week. After BB administration, increased excretion of coproporphyrins was detected only at the highest dose. The increased excretion of coproporphyrins following the administration of HBB could be observed already at the lowest dose (15 mg/kg). The excretion of uroporphyrins increased after two higher doses (75 and 375 mg/kg) in the fourth week of exposure. HBB also caused elevation of microsomal P450 level. The data suggest porphyrogenic activity of HBB; whereas in the case of BB we cannot exclude that elevated excretion of ALA-U resulted from kidney impairment. PMID- 11057698 TI - Structure dependent induction of CYP1A by polychlorinated biphenyls in hepatocytes of male castrated pigs. AB - Hepatocytes cultures prepared from castrated pig hepatocytes (Great Yorkshire x Dutch Landrace), as a model for human liver, were used to study the effect of twenty polychlorinated biphenyls (PCBs) on CYP1A activity, measured as the dealkylation of either ethoxyresorufin or methoxyresorufin. The selection of the PCBs was based on their differences in physico-chemical properties. The non-ortho and mono-ortho substituted PCBs were the most potent CYP1A inducers in pig hepatocytes. In addition, several multiple-ortho substituted congeners, with five or more chlorine atoms, were inducers of CYP1A activity as well. Their relative effect potencies (REP) were proximately 10,000 times lower than the most potent congener, 3,3',4,4',5 PeCB (PCB#126). Using partial least-squares (PLS) modeling, predictions of CYP1A activity could be made for all tetra to hepta substituted congeners. Several multiple-ortho substituted PCBs, which are highly abundant in the biotic and abiotic environment, have been found to induce CYP1A activity in pig hepatocytes. Because induction of CYP1A activity is used as biomarker for Ah receptor mediated responses, it is suggested to include these congeners in future risk assessment. PMID- 11057699 TI - Corporal punishment and primary prevention of physical abuse. AB - OBJECTIVE: To bring to the attention child maltreatment professionals the potential for primary prevention of physical abuse of ending or reducing corporal punishment by parents. METHOD: The October 1999 special issue of Child Abuse & Neglect on "A National Call to Action: Working Toward the Elimination of Child Maltreatment" was reviewed in relation to coverage of corporal punishment by parents. RESULTS: Corporal punishment was not mentioned in any of the nine articles. CONCLUSIONS: The combination of research showing that corporal punishment is a major risk factor for physical abuse and research showing the wide prevalence and chronicity of corporal punishment suggests that the "National Call For Action" should include steps to end use of corporal punishment as a mode of discipline. PMID- 11057700 TI - Child sexual abuse prevention programs: do they decrease the occurrence of child sexual abuse? AB - OBJECTIVE: The primary goal of the current study was to determine whether rates of child sexual abuse differed among undergraduate women who either had or had not participated in a sexual abuse prevention program during childhood. A secondary goal was to determine whether differences emerged in sexual satisfaction or avoidance of sexual activity between those women who had or had not participated in such a program. METHOD: Eight hundred and twenty-five women undergraduates from a New England state university filled out a survey on "sexual experiences" for research credit. Respondents were asked detailed questions regarding past histories of child sexual abuse and participation in school-based prevention programs during childhood. Additionally, they responded to questions about their current sexual satisfaction and sexual behaviors. RESULTS: Sixty-two percent of the sample reported having participated in a "good touch-bad touch" sexual abuse prevention program in school. Eight percent of respondents who reported ever having had a prevention program also reported having been subsequently sexually abused, compared to 14% of respondents who did not ever have a prevention program. No differences were found in adult sexual satisfaction or on behavioral measures of sexual activity between those respondents who had and had not participated in a prevention program. CONCLUSIONS: This is the first study to find that school-based child sexual abuse prevention programs are associated with a reduced incidence of child sexual abuse. Additionally, contrary to concerns voiced in the literature, there was no evidence that prevention programs are associated with decreased sexual satisfaction or avoidance of sex in adulthood. Implications of the results for further study are discussed. PMID- 11057701 TI - Programs for the promotion of family wellness and the prevention of child maltreatment: a meta-analytic review. AB - OBJECTIVE: The objectives were to determine the effectiveness of programs in promoting family wellness and preventing child maltreatment and to identify factors that moderate program success. METHOD: Meta-analysis, employing a 3-step model testing procedure, was used to review 56 programs designed to promote family wellness and prevent child maltreatment. RESULTS: The effect sizes for proactive interventions were larger at follow-up than at post-assessment, while the effect sizes for reactive interventions were higher at post-assessment than follow-up. The lowest effect sizes for home visitation programs on child maltreatment were for programs with 12 or fewer visits and less than a 6-month duration. Intensive family preservation programs with high levels of participant involvement, an empowerment/strengths-based approach, and a component of social support had higher effect sizes than programs without those elements. Also, both home visitation and intensive family preservation interventions achieved higher effect sizes with participants of mixed socioeconomic status (SES) than participants with low SES. CONCLUSIONS: The total mean weighted effect size was .41, indicating that outcomes for the intervention group exceed 66% of those in control/comparison groups. The findings from this review demonstrated that child maltreatment can be prevented and that family wellness can be promoted. PMID- 11057702 TI - Changes in reports and incidence of child abuse following natural disasters. AB - OBJECTIVE: The aim of this research was to investigate if there is a higher incidence of child abuse following major natural disasters. METHODOLOGY: Child abuse reports and substantiations were analyzed, by county, for 1 year before and after Hurricane Hugo, the Loma Prieta Earthquake. and Hurricane Andrew. Counties were included if damage was widespread, the county was part of a presidential disaster declaration, and if there was a stable data collection system in place. RESULTS: Based on analyses of numbers, rates, and proportions, child abuse reports were disproportionately higher in the quarter and half year following two of the three disaster events (Hurricane Hugo and Loma Prieta Earthquake). CONCLUSIONS: Most, but not all, of the evidence presented indicates that child abuse escalates after major disasters. Conceptual and methodological issues need to be resolved to more conclusively answer the question about whether or not child abuse increases in the wake of natural disasters. Replications of this research are needed based on more recent disaster events. PMID- 11057703 TI - Children's experience of violence in China and Korea: a transcultural study. AB - OBJECTIVE: There were two aims: First, to compare children's rates of being battered in home, by peers, and by teachers among students between China and Korea, and second, to identify particular risk factors for such violence. METHODS: Children in grades four through six in Shanghai (238 cases) and Yanji (245 cases) in China and Seoul (248 cases) and Kimpo (241 cases) in Korea were surveyed by questionnaire method. They were asked to complete the Straus' Conflict Tactics Scale and their frequencies in the three situations respectively, and other demographic items. RESULTS: Family violence during the last 1 year was experienced in 70.6% (minor 42.2%; serious 22.6%) of the children in China and 68.9% (minor 9.4%; serious 51.3%) of those in Korea. Experience rates of violence by peers were 42.7% (minor 25.7%; serious 13.7%) in China and 26.0% (minor 11.5%; serious 14.3%) in Korea. Finally, rates of corporal punishment by teachers were 51.1% (minor 28.0%; serious 4.1%) in China and 62.0% (minor 8.8%; serious 43.8%) in Korea. The most important and common risk factor for violence in one situation was the presence of violence in another situation. CONCLUSIONS: The findings indicate that the differences in children's overall experience rates were not particularly striking. However, Korean children experienced more severe forms of violence from family members and from teachers. Findings of risk factors clearly imply that there are children vulnerable to violence from multiple sources. PMID- 11057704 TI - Maternal behaviors associated with smothering: a preliminary descriptive study. AB - OBJECTIVES: To describe maternal behavior in 15 women identified as having smothered their children. DESIGN: A descriptive study of maternal behavior and interaction with her child, using videotapes of mother and child together. These were obtained by covert video surveillance in a hospital setting. Maternal behavior was rated using an assessment schedule designed to be used with video. RESULTS: The mothers showed a range of behaviors. Three groups emerged; one whose interaction with the child resembled normal maternal behavior, a second who interacted in a hostile way, and a third who showed a paucity of interaction. CONCLUSION: These preliminary data suggest that smothering may reflect more than one type of abnormal maternal relationship or attitude towards children. This may have implications for treatment and prognosis. PMID- 11057705 TI - Risk factors for a New Zealand sample of sexually abusive children and adolescents. AB - OBJECTIVE: The aim was to understand variables contributing to the occurrence of sexual offending in children and adolescents. METHOD: Twenty youngsters who had engaged in coercive sexual behavior involving other children were compared to a matched group of clinic-referred youngsters with conduct disorders. Assessments included family histories, interviews, and psychometric measures. RESULTS: There was no group difference in the frequency of experiencing sexual abuse. However, sexually abusive youngsters were more often exposed to adult caregivers with known histories of being sexually abusive. These children had been subjected to multiple distortions of adult attachment, tended to internalize distress, and failed to use available social supports. They were likely to have experienced physical and verbal abuse; children with behavior disorders were more likely to have a parent with a psychiatric illness and to have experienced parental conflict. CONCLUSION: Severely disrupted attachment, in conjunction with family experiences of inappropriate sexual expression, place children and adolescents at risk for sexual offending. Sexually abusive youngsters had less social supports and their offending occurred after a negative emotional experience, when there was opportunity in the form of younger victims. The study suggests an interaction among developmental pathways, coping skills, and immediate proximal variables for the perpetration of sexual abuse by young people. PMID- 11057706 TI - Social correlates and coping measures of street-children: a comparative study of street and non-street children in south-western Nigeria. AB - OBJECTIVE: This paper sought to achieve two objectives: First, to identify the social correlates attributable to street-children in south-western Nigeria as well as predisposing factors to this behavior; second, it also tried to uncover the survival mechanisms of street children. METHOD: The study was carried out in Ibadan and Lagos metropoles in south-western Nigeria. A pilot study was first conducted to clarify issues like location of the children on the streets, time of the day suitable for interview, and adequacy of the instruments. The main study involving a comparative 202 and 201 street-children and non-street children, respectively, was carried out using the questionnaire and case study approach in the two cities. Data analytical procedures involved both quantitative and qualitative methods. RESULTS: Street-children are mostly males, have low-levels of education, are predominantly Yoruba, and come from families with five or more siblings. Parents of street-children commonly had low education and were mainly found in unskilled occupations: were in contract polygynous marriages which are also often characterized by marital disruption. Street children also left home because of parental/familial reasons, chiefly among which was the inability of their parents to meet with their expectations. They survive on the streets engaging in some income-yielding activities, and are also faced with many hazards. CONCLUSIONS: The findings revealed that polygyny, large family, family disruption, and child labor were all central issues and predisposing factors to living on the streets by children. The poverty factor clearly came out as a very important factor. PMID- 11057707 TI - Child sexual abuse: a case study in community collaboration. AB - OBJECTIVE: This is an exploratory study that describes the process and outcomes of a Midwestern US community's approach to case management of child sexual abuse. METHOD: Data were abstracted from 323 criminal court files. Specific information gathered included child and suspect demographic data, law enforcement and CPS involvement, child disclosure patterns and caretaker responses, offender confession, offender plea, trial and child testimony information, and sentences received by offenders. Both case process and outcome variables were examined. RESULTS: In this community, criminal court records reflect a sex offense confession rate of 64% and a sex offense plea rate of 70%. Only 15 cases went to trial and in six the offender was convicted. CONCLUSION: Communities can achieve successful outcomes when criminal prosecution of sexual abuse is sought, but the child's testimony is not necessarily the centerpiece of a successful case. In this study, desired outcomes were a consequence of the collaborative efforts of law enforcement, CPS, and the prosecutor's office, which resulted in a high confession and plea rate. PMID- 11057708 TI - Working models about mother-child relationships in abandoned children. AB - OBJECTIVE: The main purpose of the research was to find the differences between normal and abandoned children's conceptions about mother-child relationships in positive affect attribution, sense of fairness, and locus of responsibility in punishment. METHOD: Two groups of abandoned (N = 60) and one group of non abandoned (N = 36) school-aged children were told six (kind/unkind, mother to child/child to mother) short stories, and were required to answer some questions about the reasons, reactions, and attributions of both protagonists. Children's answers were classified into categories for each question, and proportions of responses were analyzed. RESULTS: Significant differences between abandoned and non-abandoned children's response categories were found as predicted. Data indicate that abandoned children's working models could be described as showing less positive affect attribution to the mother, more compliant behavior in the child, more justification of the mother when her behavior was unfair, and less successful resources in the child to be on good terms again in the relationship. CONCLUSIONS: The results of this study indicate that abandoned children's conceptions about child-mother relationships present certain peculiarities that may contribute to their present and future difficulties in adapting successfully to their social environment and in developing a healthy sense of self respect. PMID- 11057709 TI - Is race or ethnicity a predictive factor in Shaken Baby Syndrome? AB - OBJECTIVE: Previous studies have concluded that shaken baby syndrome occurs more often among Whites than among Blacks. The purpose of this study was to determine whether race is a predictive factor in Shaken Baby Syndrome when population and referral patterns are considered. METHODS: A retrospective medical record review of closed head injuries due to child abuse during the time period January 1992 to July 1997 was conducted at three pediatric tertiary care medical centers in North Carolina. Patients included children, ages 0-4 years, identified from medical record reviews and child abuse databases. Only North Carolina residents were included. The specific rates of shaken baby syndrome in Whites versus non-Whites in the referral area were computed. RESULTS: The difference in the rate of shaken baby syndrome from the referral area was not statistically significant among Whites versus non-Whites (26.7/100,000 versus 38.6/100,000, p = .089) Most of the perpetrators were male (68%), and most victims (76%), lived with their mothers and biologic father or mother's boyfriend. CONCLUSION: Race was not a significant factor in predicting shaken baby syndrome in the referral area studied, and therefore is not a useful factor in targeting groups for intervention. PMID- 11057710 TI - Susceptibility of Pseudaletia unipuncta (Lepidoptera: Noctuidae) to entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) isolated in the Azores: effect of nematode strain and host age. AB - The armyworm, Pseudaletia unipuncta (Haworth), is a serious pest to the Azores's pastures. In laboratory bioassays we tested the susceptibility of this insect to entomopathogenic nematodes isolated in Azores: Steinernema carpocapsae Az20, Az150, and A48 strains, S. glaseri Az26 strain and Heterorhabditis bacteriophora Az33 strain. The A48, Az20, and Az150 strains caused parasitism rates of 96.6, 90, and 53.3%, and mortality rates of 63.3, 46.6, and 23.3%, respectively, to sixth instar. The Az33 strain caused a parasitism rate of 73.3% and a mortality rate of 40%; whereas, the Az26 strain caused a parasitism rate of 40% and no mortality. A linear response dose-parasitism with a positive regression (r2 = 0.993) was observed in insects exposed to S. carpocapsae Az150 strain. Positive regressions were also observed between mortality and dose rate for S. carpocapsae A48 (r2 = 0.980), Az20 (r2 = 0.956), and Az150 (r2 = 0.963) strains, and H. bacteriophora Az33 strain (r2 = 0.999). Fourth instars were the most susceptible to the A48 strain, followed by the fifth instars, while the sixth instars were the less susceptible, with LD50 values of 26.2, 62.8, and 320.7 infective juveniles, respectively. The lethal time for each of the tested instars was 32.3, 35.5, and 49.2 h, respectively. The invasion rate was 33.5, 28.2, and 40.8 nematodes per treated larvae in the fourth, fifth, and sixth instars, respectively. PMID- 11057711 TI - Effect of Beauveria bassiana and Metarhizium anisopliae (Deuteromycetes) upon the coffee berry borer (Coleoptera: Scolytidae) under field conditions. AB - The effect of three strains of the fungus Beauveria bassiana (Balsamo) Vuillemin and two strains of Metarhizium anisopliae (Metschnikoff) Sorokin upon the coffee berry borer, Hypothenemus hampei (Ferrari), was studied in three coffee farms at different altitudes (450-1,100 m above sea level) in Soconusco, Chiapas, Mexico. The maximum average percentage mycosis varied according to altitude. At 450 m asl (El Rincon) mycosis was 14.3% for B. bassiana and 6.3% for M. anisopliae; at 880 m asl (Santa Anita) mycosis was 40.6% for B. bassiana and 12.6% for M. anisopliae, and at 1,100 m asl (Alpujarras) 33.9% for B. bassiana and 22. 1% for M. anisopliae. The effect of fungal mycosis through time was not significant (P > 0.01) in any of the farms, but there was a significant difference between the strains of the fungus (P < 0.01); the best strains being Bb25 and Ma4 at the lower altitude, Bb26 and Ma4 for the middle altitude and Bb26 and Ma4 at the higher altitude. Environmental factors such as temperature, relative humidity and rain were not correlated with the percentage mycosis caused by B. bassiana and M. anisopliae. However, in the case of B. bassiana there was a significant, positive correlation (P < 0.01) between the infestation levels of the pest and the mycosis response of the entomopathogen. PMID- 11057712 TI - Dispersal patterns of pest earth mites (Acari: Penthaleidae) in pastures and crops. AB - Control methods in pest earth mites and other mites often depend on low dispersal rates, yet there are no experimental estimates of these rates. To rectify this, adult movement rates were estimated in the earth mite Halotydeus destructor Tucker and the winter grain mite, Penthaleus major (Duges), using mark-release recapture techniques. Mean square dispersal distances were used to estimate diffusion coefficients. In pasture, coefficients were in the range 0.3-1.3 for these species. This suggests that 90% of the population moves < 5-11 m in a 10-d period, or 7-16 m within their adult lifetime. Releases of mites in adjacent pea/wheat crops indicated directional movement toward the more favored pea host. However, there was no directional movement when adjacent plots of peas and lupins were compared, even though lupins are poor hosts. These results indicate that broad border sprays or border culturing will be needed to prevent mite movement from adjacent paddocks. PMID- 11057713 TI - Multiacreage evaluation of aerially applied adherent malathion granules for selective insect control and indirect reduction of mycotoxigenic fungi in specialty corn. AB - Aerially applied adherent corn flour granules containing 1% malathion were more often as, or more, effective than 15% chlorpyrifos (Lorsban 15G) granules in controlling caterpillars and sap beetles in high amylose corn in 1997 than 1996. Use of malathion granules corresponding closely in size to chlorpyrifos granules in the second year of the study apparently increased relative efficacy. Significantly less corn borer damage occurred on plants (1996) or ears (1997) within 2 wk of application for both types of insecticide granules compared with untreated plots. In 1997, there were sixfold fewer milk stage ears with more than 20 kernels damaged per ear in the malathion-treated plots compared with chlorpyrifos-treated plots, and severity of caterpillar damage was also less in malathion versus chlorpyrifos-treated plots at harvest. Control of beetles (corn rootworm adults and sap beetles) for both treatments was less effective compared with caterpillars. Significant corn rootworm adult control was noted for both chlorpyrifos and malathion in 1996 and significant sap beetle control was noted for the malathion granules in 1997. Significantly fewer live lady beetles, and more dead lady beetles were present in chlorpyrifos-treated plots compared with malathion-treated or untreated plots in 1996. The incidence and severity of Fusarium mold on ears at harvest was often indirectly reduced by both malathion treatments and chlorpyrifos treatments, with the malathion treatment significantly better than the chlorpyrifos treatment in one case. PMID- 11057714 TI - Simulated insect defoliation on soybean: influence of row width. AB - An ongoing change in soybean production gaining popularity in the United States is a reduction in row spacing. Plant canopy closure is quicker and leaf area index is greater, thus yield is usually higher. Because yield response to insect defoliation is primarily a function of how defoliation causes changes in light interception, the possibility exists that the insect-injury-yield-loss relationship might differ among row widths. Soybean was grown in four states using similar methodologies. Insect defoliation was simulated by picking leaflets based on an insect defoliation model. Plant growth measurements were taken immediately following the end of defoliation. Numerous independent variables were measured or calculated, including percentage light interception, leaf area index, percentage defoliation, and leaf area per plot. Analyses of covariance were conducted on the resultant data to determine whether insect-injury-yield-loss relationships interact with row width. A significant interaction would indicate that the impact of the variables on yield was dependent on the row width, whereas a nonsignificant interaction would suggest that the relationship between the variables and yield is similar at all row widths. Few significant interactions were obtained, indicating that the impact of the variables on yield is similar across row widths. Because of the lack of significant interactions, the insect injury-yield-loss relationships previously developed should be usable across varying row widths. Thus, treatment decisions based on light interception and leaf area indices, both considered more appropriated measures of insect injury, should be applicable for all row spacings. PMID- 11057715 TI - Effects of weed management systems on canopy insects in herbicide-resistant soybeans. AB - The effects of transgenic herbicide-resistant soybean varieties and their corresponding weed management strategies on canopy insects were examined in studies at two locations in Iowa in 1997 and 1998. Weed management systems that allowed more weed escapes typically had higher insect population densities. However, systems with fewer weeds seemingly were preferred by potato leafhoppers. Bean leaf beetles and potato leafhoppers showed preferences for certain soybean varieties, but these effects were attributed to soybean plant height. These findings indicate that although the transgenic soybean varieties did not strongly affect insect populations, weed management systems can affect insect populations in soybean. However, this impact is likely related more to weed suppression effectiveness than to a direct effect of the herbicides on the insects. PMID- 11057716 TI - Impact of Leptoglossus occidentalis (Hemiptera: Coreidae) on Douglas-fir seed production. AB - We investigated the effect of feeding by the western conifer-seed bug, Leptoglossus occidentalis Heidemann, on seed production in developing cones of coastal Douglas-fir, Pseudostuga menziesii (Mirb.) Franco, with respect to seed bug life stage and sex (nymphs, adult females, and adult males) and timing of feeding (early, mid-, and late season cone development). Feeding by females on caged cones for a 2-wk period during late season cone development reduced the proportion of full seeds in cones by approximately 70% compared with caged control cones. There was no significant difference among nymphs, adult females, and adult males with respect to the proportion of empty or partially fed-upon seeds produced during the same feeding period. Feeding by nymphs for 2 wk early in the season resulted in a threefold increase in the number of unextractable seeds fused to cones compared with the control. Weight measurements of harvested seeds indicated that radiography is an accurate tool to distinguish among Douglas fir seeds that have sustained light, moderate, or severe damage. Determining the full impact of L. occidentalis on conifer seed production will require the development of a reliable method to distinguish between naturally aborted seeds and seeds emptied through feeding by seed bugs. PMID- 11057717 TI - Effect of trap size, placement, and age on captures of blueberry maggot flies (Diptera: Tephritidae). AB - Ammonium acetate and protein hydrolysate baited and unbaited green spheres (3.6, 9.0, and 15.6 cm diameter) were evaluated for effectiveness in capturing blueberry maggot flies, Rhagoletis mendax Curran. Early in the season, baited spheres (9.0 cm diameter) captured significantly more R. mendax flies than spheres of 3.6 and 15.6 cm diameter. As the season progressed, the differences in trap captures became less pronounced among the 3.6-, 9.0-, and 15.6-cm-diameter spheres. In other experiments, the effects of trap positions and age on captures of blueberry maggot flies were assessed. Traps were positioned 15 cm above the bush canopy, 15 cm inside the canopy (from top of the bush), and 45 cm from the ground. Traps placed within the canopy captured 2.5 and 1.5 times as many flies compared with traps placed above the canopy and 45 cm from the ground, respectively. When sticky yellow Pherocon AM boards and green sphere traps were allowed to age in field cages, freshly baited (0 d) yellow sticky boards captured significantly more blueberry maggot flies than boards aged for 11, 28, and 40 d, respectively. No significant differences were observed among boards aged for 11, 28, and 40 d. However, when baited 9-cm sticky spheres were aged in field cages, there were no significant differences between freshly baited spheres and spheres aged for 11 and 28 d, respectively. Spheres aged for 40 d differed significantly from freshly baited ones. The study demonstrated that the baited 9-cm-diameter sphere was more effective in capturing blueberry maggot flies than spheres of 3.6 and 15.6 cm diameter. When this trap is deployed in the center of the bush canopy approximately 15 cm from the top of the bush, it is attractive and accessible to R. mendax flies. The data also indicated that a baited 9-cm sphere has a longer effective life span than Pherocon AM boards when deployed under the same field conditions. PMID- 11057718 TI - Particle film deters oviposition by Diaprepes abbreviatus (Coleoptera: Curculionidae). AB - A hydrophilic formulation of the inert silicate kaolin was tested in a screenhouse for its effect on the behavior of the root weevil Diaprepes abbreviatus (L.), a pest of citrus and ornamental plants in Florida and the Caribbean. Feeding by adults on treated foliage was reduced by 68-84% compared with adults fed untreated foliage. No insecticidal activity was detected after 14 d of exposure to kaolin-treated leaves. Oviposition was completely suppressed on treated foliage. Although females oviposited >19,000 eggs during two trials on untreated foliage, no egg masses were found on foliage treated with the kaolin formulation. These data indicate potential for kaolin as a barrier to oviposition in citrus groves and may prove to be an economically viable and environmentally sound component of an integrated approach for control of D. abbreviatus and related root weevils. PMID- 11057719 TI - Management strategy, shade, and landscape composition effects on urban landscape plant quality and arthropod abundance. AB - Intensity and type of management, the cultural variable shade, and the combination of woody and herbaceous annual and perennial plants were evaluated for their effect on key landscape arthropod pests. Azalea lace bugs, Stephanitis pyrioides (Scott), and twolined spittlebugs, Prosapia bicincta (Say), were most effectively suppressed in landscape designed with resistant plant species of woody ornamentals and turf. Landscapes containing susceptible plant counterparts were heavily infested by these two insect species in untreated control plots. A traditional management program of prescribed herbicide, insecticide, and fungicide applications effectively suppressed azalea lace bug and produced a high quality landscape. Targeted integrated pest management with solely horticultural oils resulted in intermediate levels of azalea lace bug. Neither program completely controlled twolined spittlebug on hollies or turf. Carabidae, Staphylinidae, Formicidae, and Araneae were not reduced by any management strategy. Lace bugs (Stephanitis) were more common in plots with 50% shade than those in full sun. Spittlebugs (Prosapia) were more common in the shade during 1996 and in the sun during 1997. Spiders and ants were more often collected in full sun plots. Carabids, staphylinids, and spiders were more commonly collected from pitfall traps in turf than in wood-chip mulched plant beds, whereas ants were equally common in both locations. The addition of herbaceous plants to the landscape beds had little effect on pest insect abundance. PMID- 11057720 TI - Effectiveness of thiamethoxam-coated spheres against blueberry maggot flies (Diptera: Tephritidae). AB - Studies were conducted to evaluate the mortality of blueberry maggot, Rhagoletis mendax Curran, flies exposed to thiamethoxam- and imidacloprid-coated biodegradable (8-cm-diameter) red spheres, under both laboratory and field conditions. Laboratory studies with spheres coated with 0.1-2.0% (AI) of thiamethoxam indicated that they are effective against R. mendax; however, no dose-dependent response was observed. Studies on the effect of visitation time on thiamethoxam-coated spheres showed a decrease in R. mendax mortality as the duration of visitation time decreased from 60 to 10 s. Under field conditions, significantly more flies were captured on Plexiglas panes below the 2% (AI) thiamethoxam-coated spheres when compared with similar panes below untreated spheres. In field evaluations of thiamethoxam- and imidacloprid-coated spheres, imidacloprid-coated spheres (2.0% [AI]) were found to be significantly more effective than thiamethoxam-coated spheres (0.5-4.0% [AI]). Field trials to characterize the levels of mortality associated with aging pesticide-coated spheres revealed that the effectiveness of treated spheres decreased with increasing age of sphere, and this reduction in effectiveness is greater in thiamethoxam-coated spheres than in imidacloprid-coated spheres. These results provide comparative data on the effectiveness of thiamethoxam- and imidacloprid coated spheres and support the potential of using pesticide-treated spheres for control of blueberry maggot flies. PMID- 11057721 TI - Development of an attractant for the scarab pest Macrodactylus subspinosus (Coleoptera: Scarabaeidae). AB - Field trials were conducted over several seasons to determine the attractant most successful in luring adult rose chafers, Macrodactylus subspinosus (F.), to traps. During the first season, 20 compounds were compared with the standard lure, valeric acid + hexanoic acid + octyl butyrate (1:1:1). The two new standards establishedthat season were: valeric acid + 1-nonanol (1:1); and valeric acid + hexanoic acid + octyl butyrate + 1-nonanol (1:1:1:1). The following season, 36 compounds were evaluated, comparing them to the new standards. The performance of the standard binary lure valeric acid + 1-nonanol was improved when the alcohol 1-nonanol was replaced by its analog trans-2 nonenol and this was confirmed during the third season. At the same time, a second test was conducted with 29 new candidates, which were combined with valeric acid and compared with the standard: valeric acid + hexanoic acid + octyl butyrate + 1-nonanol (1:1:1:1). A control and the single compound alpha-ionone were included, resulting in the discovery of a new more powerful attractant, alpha-ionone. Testing of alpha-ionone continued the following season, at which time the initial leading candidate and new ones containing trans-2-nonenol were tested against the single attractant alpha-ionone and various combinations of it. A new five-component mixture of valeric acid, hexanoic acid, octyl butyrate, trans-2-nonenol, and alpha-ionone out performed all other lure combinations. PMID- 11057722 TI - Sodium tetraborate effects on mortality and reproduction of Anastrepha suspensa (Diptera: Tephritidae). AB - When flies were treated with 0- 0.5% sodium tetraborate by feeding for 24 h, mortality in treatments was not different from controls. Fecundity and fertility were reduced by 0.5% sodium tetraborate. When flies were fed for 48 h, mortality of both males and females increased in the 0.5% sodium tetraborate treatment; oviposition was eliminated for 20 d after treatment. When treatment was extended to 168 h, 0.1% sodium tetraborate caused increased mortality and decreased fecundity and fertility. Fed for 168 h, 0.2 and 0.5% sodium tetraborate killed almost all flies within the 7-d treatment. Oviposition of survivors in 0.1 and 0.2% sodium tetraborate treatments was arrested for 20 d after treatment. PMID- 11057723 TI - Evaluation of control measures for black carpenter ant (Hymenoptera: Formicidae). AB - Current control methods for the black carpenter ant, Camponotus pennsylvanicus (De Geer), include the use of remedial and preventative residual sprays as well as toxic baits. We evaluated the acceptance of three baits (Maxforce, Niban, and Baygon) to field colonies of the black carpenter ant in the spring and fall. Maxforce bait granules were more readily accepted than either Niban or Baygon bait granules in the spring. A change in food preference from protein to sugar by the black carpenter ant appeared to reduce the number of Maxforce bait granules removed in the fall, resulting in no differences in bait acceptability. The longevity of Dursban 50W and Tempo 20WP were evaluated in the summer and fall on painted wood panels. Panels aged outside for 15 d under prevailing weather conditions exhibited increased LT50 values. For each sampling period, panels aged on the south face (in the sun) exhibited less insecticidal activity (i.e., large LT50 values) than panels on the north face (shaded; small LT50 values). At each sampling period, Tempo 20WP provided smaller LT50 values than Dursban 50W. Because of changing dietary preferences, our data highlight the importance of using various bait types for carpenter ant control. Moreover, the application of residual spays should be made to locations protected from direct sunlight. PMID- 11057724 TI - Response of Reticulitermes spp. (Isoptera: Rhinotermitidae) in northern California to baiting with hexaflumuron with sentricon termite colony elimination system. AB - Colonies of Reticulitermes spp. were baited with prototype and commercial Sentricon stations (Dow AgroSciences LLC, Indianapolis, IN) to test the efficacy of hexaflumuron in different concentrations and bait matrices and to document reinvasion of the foraging territories vacated by eliminated colonies. Seven colonies of Reticuliternes spp. from two sites were characterized with cuticular hydrocarbon analyses and mark-release-recapture and agonistic behavioral studies. Three colonies were observed as controls and four colonies were baited. When a connection between the bait station and the monitoring station could not be confirmed by mark-release-recapture studies, the results of the baiting were equivocal. The monitoring stations of a colony at our wildland site were devoid of termites 406 d after baiting with one Sentricon station, but became reoccupied with the same species of termites approximately 6 mo after baiting. A colony at the residential site was baited with 0.5% hexaflumuron in the Recurit II bait matrix; 60 d later termites were absent from all monitoring stations. These monitoring stations remained unoccupied for > or = 18 mo. Foraging Reticulitermes spp. appeared in three of the seven monitoring stations 18, 24, and 36 mo after baiting, respectively. Using cuticular hydrocarbon analyses and agonistic behavior studies, we determined that the Reticulitermes spp. occupying these monitoring stations were from three different colonies; none were members of the original colony destroyed by baiting. Another colony at the residential site was baited using a noncommercial, experimental bait; 52 d later termites were absent from all monitoring stations. The monitoring stations remained unoccupied for > or = 9 mo. A different Reticulitermes sp. colony invaded one monitoring station 9 mo after baiting. PMID- 11057725 TI - Inheritance and synergism of resistance to imidacloprid in the Colorado potato beetle (Coleoptera: Chrysomelidae). AB - Colorado potato beetle, Leptinotarsa decemlineata (Say), adults and larvae collected from Long Island, NY, were 100.8 and 13.2 times more resistant to imidacloprid, respectively, compared with a susceptible strain. This high level of resistance appeared in only the third field season of imidacloprid use. Analysis of probit lines from F1 reciprocal crosses indicated that resistance to imidacloprid in adults was inherited autosomaly as an incompletely recessive factor. The degree of dominance of the resistance was -0.23 and -0.10, respectively, 3 and 7 d after treatment (incompletely recessive). The chi2 analysis of response ratio statistics from F1 x susceptible back crosses compared with a monogenic model suggested that more than one locus is responsible for resistance to imidacloprid. Synergism studies with piperonyl butoxide suggested that mixed-function oxidase mediated detoxification is responsible for the resistance to imidacloprid in adults. Synergism studies with S,S,S-tributyl phosphorotrithioate (DEF) indicated that esterase mediated detoxification may be an additional resistance factor. Mixed-function oxidase mediated detoxification is probably also one of the mechanisms of resistance to imidacloprid in larvae. Because the synergists used did not completely eliminate resistance in the resistant strain, there may be additional mechanisms involved. Refugia and crop rotation decrease the frequency of homozygous resistant genotypes and may be effective resistance management strategies, because of the recessive nature of the resistance. PMID- 11057726 TI - Frequency of alleles conferring resistance to a Bacillus thuringiensis toxin in a Philippine population of Scirpophaga incertulas (Lepidoptera: Pyralidae). AB - Using the F2 screen methodology, we estimated the frequency of alleles conferring resistance to the Cry1Ab toxin of Bacillus thuringiensis Berliner in a Philippine population of the stem borer Scirpophaga incertulas (Walker). Evaluation of >450 isofemale lines for survival of F2 larvae on cry1Ab plants did not detect the presence of an allele conferring a high level of resistance. The frequency of such an allele in the sampled population was conservatively estimated to be <3.6 x 10(-3) with 95% confidence and a detection probability of 94%. However, there was evidence of the presence of alleles conferring partial resistance to Cry1Ab. The frequency of alleles for partial resistance was estimated as 4.8 x 10(-3) with a 95% CI between 1.3 x 10(-3) and 1.04 x 10(-2) and a detection probability of 94%. Our results suggest that the frequency of alleles conferring resistance to Cry1Ab in the population of S. incertulas sampled is not too high to preclude successful implementation of the high dose/refuge resistance management strategy. PMID- 11057727 TI - Development and validation of a binomial sequential sampling plan for the greenbug (Homoptera: Aphididae) infesting winter wheat in the southern plains. AB - From 1997 to 1999, Schizaphis graminum (Rondani), intensity (number per tiller) was estimated on 115 occasions from hard red winter wheat fields located throughout the major wheat growing regions of Oklahoma. A total of 32 and 83 fields was sampled during the fall and spring, respectively. The parameters of linear regressions relating the mean number of greenbugs per tiller (m) and the proportion of infested tillers (PT) differed significantly between fall and spring infestations. The PT-m linear model provided a good fit for data on S. graminum for fall and spring infestations at tally thresholds of 0, 1, 2, and 3. A tally threshold (T) represents the number of greenbugs present on a tiller before the tiller is classified as infested by >T greenbugs. A regression model with a tally threshold of 2 was the most precise for classifying S. graminum populations during fall growth of winter wheat because it explained a greater amount of the variation in the PT-m relationship (97%) than models with other tally thresholds. A separate spring model with a tally threshold of 1 was the most precise for classifying S. graminum populations during spring growth of winter wheat. Sequential sampling stop lines based on sequential probability ratio tests were calculated for economic thresholds of 3 or 6 greenbugs per tiller for fall infestations and 6 or 9 greenbugs per tiller for spring infestations. With the newly developed parameters, the average sample number required to classify greenbug populations near economic thresholds (as above or below the economic threshold) varied from 69 to 207. We expect that the sampling plans for greenbugs in winter wheat developed during this study will be efficient and useful tools for consultants and producers in the southern plains. PMID- 11057728 TI - Monitoring insect pests in retail stores by trapping and spatial analysis. AB - Stored-product insects are a perennial problem in retail stores, where they damage and contaminate susceptible merchandise such as food products and animal feed. Historically, pest management in these stores has relied heavily on chemical insecticides, but environmental and health issues have dictated use of safer methods, and these require better monitoring. A monitoring procedure that employs an array of moth and beetle traps combined with spatial (contour) analysis of trap catch was tested in three department stores and two pet stores. The rate of capture increased with the level of infestation but was essentially constant over 4- to 5-d trapping periods. Contour analysis effectively located foci of infestation and reflected population changes produced by applications of the insect growth regulator (S)-hydroprene. The most abundant insects were Plodia interpunctella (Hiibner), Lasioderma serricorne (F.), Oryzaephilus mercator (Fauvel), Tribolium castaneum (Herbst), and Cryptolestes pusillus (Schonherr). The results indicate that contour analysis of trap counts provides a useful monitoring tool for management of storage pests in retail stores. It identifies trouble spots and permits selection, timing, and precision targeting of control measures to achieve maximum pest suppression with minimum pesticide risk. It permits managers and pest control operators to visualize pest problems over an entire store, to monitor changes over time, and to evaluate the effectiveness of control intervention. The contour maps themselves, along with records of control applications and stock rotation, provide permanent documentation of pest problems and the effectiveness of pest management procedures. PMID- 11057729 TI - Youth tobacco surveillance--United States, 1998-1999. AB - PROBLEM/CONDITION: Tobacco use is the single leading preventable cause of death in the United States, accounting for approximately, 430,000 deaths each year. The prevalence of cigarette smoking nationwide among high school students increased during the 1990s, peaking in 1996-1997, then began a gradual decline. Approximately 80% of tobacco users initiate use before the age of 18 years. If the trend in early initiation of cigarette smoking continues, approximately 5 million children aged <18 years who are living today will die prematurely as adults because they began to smoke cigarettes during adolescence. The economic liability associated with tobacco use ranges from $50 billion to $73 billion per year in medical expenses alone. Because of these health and economic consequences, CDC has recommended that states establish and maintain comprehensive tobacco control programs to reduce tobacco use among youth. REPORTING PERIOD: February 1998 through December 1999. DESCRIPTION OF THE SYSTEM: To assist states in developing and maintaining their state-based comprehensive tobacco prevention and control programs, CDC developed the Youth Tobacco Surveillance and Evaluation System, which includes international, national, and state school-based surveys of middle school and high school students. Two components of this system are discussed--the National Youth Tobacco Survey and the state Youth Tobacco Surveys. The national survey is representative of students in the 50 states and the District of Columbia; 15,061 students in 131 schools completed questionnaires in 1999. The state surveys were first conducted in 1998, when three states participated, and in 1999, when 13 states participated (13 states conducted middle school surveys and 10 states conducted high school surveys); state sample sizes ranged from 452 to 15,478 students. This report summarizes data from the 1999 national survey and the 1998 and 1999 state surveys. RESULTS AND INTERPRETATION: Findings from the National Youth Tobacco Survey show current tobacco use ranges from 12.8% among middle school students to 34.8% among high school students. Cigarette smoking is the most prevalent form of tobacco used, followed by cigars and smokeless tobacco. Young people have strong cigarette brand preferences. Almost half of current smokers in both middle school and high school report that they usually smoke Marlboro cigarettes. Black students are more likely to smoke Newport cigarettes than any other brand. Half of current smokers in middle school and high school report that they want to completely stop smoking. Nearly one fourth of middle school and high school students who have never smoked cigarettes indicate that they are susceptible to initiating smoking in the next year. Environmental tobacco smoke exposure is very high among both middle school and high school students. During the week before the survey, approximately 9 out of 10 current smokers and half of never smokers were in the same room with someone who was smoking; 8 out of 10 current smokers and 3 out of 10 never smokers rode in a car with someone who was smoking. Six out of 10 current smokers and 3 out of 10 never smokers live in a home where someone else smokes cigarettes. Approximately 70% of middle school and 60% of high school students who currently smoke and are aged <18 years were not asked to show proof of age when they purchased cigarettes. Approximately three fourths of middle school and high school students have seen antismoking commercials; however, 90% report having seen actors smoking on television or in the movies. Approximately 2% of middle school and high school students who had never used tobacco would wear or use something with a tobacco company name or picture on it. This rate increases to approximately 20% for current tobacco users. ACTIONS TAKEN: Youth Tobacco Survey data are used by health and education officials to improve national and state programs to prevent and control youth tobacco use. (ABSTRACT TRUN PMID- 11057730 TI - Report of the Consensus Conference on the Diagnosis of Auditory Processing Disorders in School-Aged Children. PMID- 11057731 TI - Speech recognition ability in noise and its relationship to perceived hearing aid benefit. AB - Hearing-impaired listeners with similar hearing losses may differ widely in their ability to understand speech in noise. Such individual susceptibility to noise may explain why patients obtain varying degrees of benefit from hearing aids. The chief purpose of this study was to determine if adaptive measures of unaided speech recognition in noise were related to hearing aid benefit. Additionally, the relationship between perceived hearing handicap and benefit from amplification was explored. Before being fit with hearing aids, 47 new hearing aid users completed a self-assessment measure of hearing handicap Then, unaided speech recognition ability was measured in quiet and in noise. Three months later, subjects completed a hearing aid benefit questionnaire. A weak relationship was observed between perceived hearing handicap and hearing aid benefit. There were no significant relationships between speech-in-noise measures and hearing aid benefit, suggesting that speech recognition ability in noise is not a major determinant of the benefit derived from amplification. PMID- 11057732 TI - A case of Susac syndrome. AB - Susac syndrome is a readily recognized but often misdiagnosed disorder almost exclusively affecting women in the 20- to 40-year age range. Characterized by the clinical triad of encephalopathy, branch retinal artery occlusions, and sensorineural hearing loss, patients with Susac syndrome are often misdiagnosed with multiple sclerosis (MS). Unlike MS, however, the disease process extends over a 1- to 2-year period and then goes into remission. This presentation describes the progression of symptoms of a patient eventually diagnosed with Susac syndrome. PMID- 11057733 TI - The question of phonetic balance in word recognition testing. AB - Twenty subjects with normal hearing and 15 subjects with mild-to-moderate sensorineural hearing losses were tested with eight lists of words using monosyllabic pronunciation to determine word recognition scores. Four of the lists were taken from Northwestern University Test No. 6 and four were simply made up by randomly selecting words from a dictionary. All of the word lists were used to determine performance-intensity functions. No clinically meaningful differences were observed among the lists. PMID- 11057734 TI - Investigation of binaural interference in normal-hearing and hearing-impaired adults. AB - When speech recognition testing is performed under diotic conditions, some elderly persons with asymmetric hearing loss exhibit a phenomenon in which the performance of the poorer ear interferes with that of the better ear. This binaural interference phenomenon has been estimated to occur in 8 to 10 percent of elderly hearing aid users. The purpose of the present study was to investigate the prevalence of this phenomenon in groups (n = 12) of young and elderly listeners with normal hearing, plus groups of elderly listeners with hearing loss who were aided or unaided. Of 48 subjects tested, only 2 exhibited significant evidence of binaural interference, a result that is close to that expected by chance. Although both of these subjects were elderly, one had normal hearing and the other was aided binaurally. A third elderly unaided subject exhibited a significant binaural advantage. Further studies are needed to determine the prevalence of binaural interference in normal-hearing or hearing-impaired listeners in any decade of life. PMID- 11057735 TI - Auditory temporal processing and lexical/nonlexical reading in developmental dyslexics. AB - Relationships between lexical/nonlexical reading and auditory temporal processing were examined. Poor nonlexical readers (poor nonword readers, phonologic dyslexics) had difficulty across tone tasks irrespective of speed of presentation or mode of recall. Poor lexical readers (poor irregular word readers, surface dyslexics) had difficulty recalling tones in a sequence only when they were presented rapidly. Covariate analysis supported these findings, revealing that nonlexical (nonword) reading performance is associated with general auditory performance, but lexical (irregular word) reading is particularly associated with auditory sequencing. These findings suggest that phonologic and surface dyslexics perform differently on nonspeech auditory tasks. Because the two different types of poor readers did not differ significantly on tests of memory and learning but did differ on auditory tasks, we suggest that their performance on the auditory tasks may reflect auditory processing abnormalities as opposed to more general learning or memory difficulties. In addition to these observed qualitative differences between groups on the tone tasks, collapsing groups (all readers) revealed significant correlations between nonword reading and the Same-Different tone tasks in particular, whereas irregular word reading was not associated with any tone tasks; there also appears to be a quantitative relationship between nonlexical reading and Same-Different tone task performance as better or worse nonword reading predicts better or worse performance on the Same-Different tone tasks. In particular, it is conceivable that an auditory temporal processing deficit might contribute to poor nonword reading. PMID- 11057736 TI - Acquired bilateral peripheral vestibular system impairment: rehabilitative options and potential outcomes. AB - Acquired bilateral vestibular impairment can be a devastating disorder that is most frequently the result of aminoglycoside-induced toxicity. The presenting complaints are typically oscillopsia and gait and balance disturbances. These patients can be excellent candidates for vestibular rehabilitation therapy that focuses on facilitating maximal use of any remaining vestibular function, improving gaze and postural stability through the use of visual and somatosensory cues, and improving home and workplace safety. The prognosis for recovery is determined by the extent of the loss and the presence of other progressive disorders that may affect vision or somatosensation, coexisting illnesses, and the patient's compliance with the therapy program. Two cases are presented to illustrate the salient aspects of vestibular rehabilitation for patients with acquired bilateral vestibular system loss, including factors affecting patient progress and final outcome. PMID- 11057737 TI - Radiobiological studies on the 65 MeV therapeutic proton beam at Nice using human tumour cells. AB - PURPOSE: To determine the relative biological effectiveness (RBE) for initial and delayed inactivation of cells by a modulated proton beam suitable for the treatment of tumours of the eye, within the spread-out Bragg peak and in its distal declining edge. MATERIALS AND METHODS: Human tumour SCC25 cells were irradiated with the 65 MeV proton beam at the Cyclotron Medicyc in Nice. Perspex plates of different thickness were used to simulate five positions along the beam line: 2mm corresponding to the entrance beam; 15.6 and 25 mm in the spread-out Bragg peak; 27.2 and 27.8mm for the distal edge. At each position clonogenic survival of the irradiated cells and of their progeny were determined at various dose values. 60Co gamma-rays were used as reference radiation. RESULTS: RBE values evaluated at the survival level given by 2 Gy of gamma-rays increased with increasing depth from close to 1.0 at the proximal to about 1.2 at the distal part of the peak. Within the declining edge it reached the value of about 1.4 at 27.2 and about 2 at 27.8 mm. For the progeny of irradiated cells, the RBE value ranged from 1.0 to 1.1 within the spread-out Bragg peak and then increased up to a value of 2.0 at the last position. The dose-effect curves for the progeny always had a larger shoulder than for the irradiated progenitors, their alpha parameters being lower by a factor of about 4 and their beta parameters always being higher. The alpha/beta ratio was about 50 Gy for the progenitors and about 6 Gy for their progeny. The incidence of delayed effects increased with dose and with the depth within the beam. CONCLUSIONS: RBE values for the inactivation of cells irradiated in the spread-out Bragg peak are compatible with the value currently assumed in clinical applications. In the distal declining edge of the beam, the RBE values increased significantly to an extent that may be of concern when the region of the treatment volume is close to sensitive tissues. The yield of delayed reproductive cell death was significant at each position along the beam line. PMID- 11057738 TI - Mechanism of DNA damage by thiocyanate radicals. AB - PURPOSE: It was previously shown that gamma-irradiation of aqueous solutions of plasmid DNA in the presence of millimolar concentrations of thiocyanate ions leads to the formation in very high yields of sites recognized by the base excision repair endonuclease formamido-pyrimidine-DNA N-glycosylase (FPG). The authors wished to characterize the mechanism responsible for the production of these FPG-sensitive sites. MATERIALS AND METHODS: An aqueous solution of plasmid DNA containing thiocyanate ions was irradiated with 137Cs gamma-rays. After irradiation, aliquots were treated with FPG. Break yields were determined using neutral agarose gel electrophoresis. RESULTS: The yield of FPG-sensitive sites decreased with decreasing enzyme activity, increasing thiocyanate concentration, increasing dose-rate, increasing ionic strength, increasing nitrite or iodide concentration, and decreasing oxygen concentration. CONCLUSION: The observations suggest that the monomeric thiocyanate radical SCN* is an intermediate in the reaction, and that the yields of FPG-sensitive sites are determined by competition between the disproportionation of the dimeric radical anion (SCN)*2- and the fate of a one-electron oxidized guanine species in DNA. The latter can react with oxygen to produce an FPG-sensitive site or can be reduced without producing an FPG-sensitive site. The results help to clarify the mechanisms responsible for DNA damage by the direct effect of ionizing radiation. PMID- 11057739 TI - Survival of human lung epithelial cells following in vitro alpha-particle irradiation with absolute determination of the number of alpha-particle traversals of individual cells. AB - PURPOSE: To throw light on human exposure to domestic radon and radon progeny, the effects of low doses of alpha-particle irradiation on normal human lung epithelial cells has been studied. At such low exposure levels the concept of dose is inadequate due to the stochastic variation in the number of alpha particle traversals per cell. The objective of the current study was to establish an accurate survival curve for human lung epithelial cells with absolute determination of the exact number of alpha-particle traversals of individual cells. MATERIALS AND METHODS: Irradiation of L132 cells growing in tracketch detector-based cell dishes, was performed using a collimated alpha-particle beam from a 210Po source. The number of alpha-particle traversals through each individual cell was scored by using a technique of retrospective track-etch dosimetry. This technique is based upon image matching and mapping of corresponding cell and alpha-particle track images. The spatial resolution of the hit determination procedure was +/-0.9/microm. RESULTS: Surviving fractions of cells (SF) showed strict dependence on the number of nuclear traversals (n), with SF(n)= a exp(-bn), a=0.957 (+/- 0.046), b = 0.587 (+/- 0.059), R2 =98.8%. No significant dependence on the number of nuclear membrane traversals (m) or the number of cytoplasm traversals (c) was observed. PMID- 11057740 TI - Chronic exposure to bryostatin-1 increases the radiosensitivity of U937 leukaemia cells ectopically expressing Bcl-2 through a non-apoptotic mechanism. AB - PURPOSE: Ionizing radiation (IR) produced a dose-dependent increase in apoptosis in U937/pCEP4 cells which was attenuated by the stable over expression of Bcl-2 (U937/Bcl-2). A dose of 2 Gy IR was selected for further analyses to determine if subsequent exposure to 10nM bryostatin- would overcome the resistance to IR induced apoptosis conferred by Bcl-2 over expression. METHODS AND RESULTS: Although bryostatin- did not increase IR-induced apoptosis in U937/pCEP4 or U937/Bcl-2 cells, it impaired mitochondrial function and increased the antiproliferative effects of IR in both cell lines. The effects were more pronounced in U937/Bcl-2 cells. Bryostatin-1 also exerted differential effects on cell-cycle distributions of U937 transfectant cells, producing a significant G0/G1 arrest in U937/Bcl-2 cells, while decreasing IR-induced G2/M arrest in U937/pCEP4 cells. Although Bcl-2 over expression attenuated IR-induced apoptosis, clonogenic survival was similar in U937/pCEP4 and U937/Bcl-2 cells following 2 Gy IR treatment. Treatment with 10nM bryostatin-1 after 2 Gy IR further reduced clonogenic survival in both cell lines. Moreover, U937/Bcl-2 cells were more susceptible to the growth-inhibitory effects of IR/bryostatin-1 than U937/pCEP4 cells. CONCLUSIONS: Bryostatin-1 increased the radiosensitivity of U937 transfectant cell lines without enhancing apoptosis; furthermore, U937/Bcl-2 cells were more susceptible to IR/bryostatin-1-mediated antiproliferative effects than their empty-vector counterparts. PMID- 11057741 TI - Effect of heat on dsb repair in G1- and S-phase studied in the human HeLa S3 cell line. AB - PURPOSE: The aim of this study was to investigate the relation between double strand breaks and thermal radiosensitization in dependence on cell-cycle position. MATERIALS AND METHODS: The experiments were performed with the human tumour cell line HeLa S3. Cells synchronized in G1- and S-phase were exposed to X rays alone or in combination with prior heating at 44 degrees C for 20 min. Cell kill was determined by means of colony forming assay, double-strand breaks (dsb) using constant-field gel electrophoresis and apoptotic cell death was scored using the fraction of detached cells. RESULTS: In both cell-cycle phases heating at 44 degrees C for 20 min prior to irradiation resulted in an increased cellular radiosensitivity, whereby the thermal enhancement ratio (TER) was significantly higher in S- than in G1-phase cells with TER=2.1 and 1.2, respectively. Prior heating at 44 degrees C did not affect the number of radiation-induced dsb but was found to modify their repair as measured for a X-ray dose of 40 Gy. In both cell cycle phases dsb repair kinetics measured after irradiation alone could be described by a fast and a slow component with the majority of dsb being repaired with fast kinetics. Prior heating at 44 degrees C was found to have only a minor effect on these half-times but mainly to affect the number of slowly rejoined dsb. In G1-phase cells the number of slowly rejoined dsb measured 300 min after irradiation was enhanced by a factor of 1.8 and in S-phase cells even by a factor of 3.2. Fraction of apoptotically dying cells was low after X-irradiation alone but was clearly enhanced after combined treatment, which was especially pronounced for S-phase cells. CONCLUSIONS: The pronounced thermal radiosensitization found for S-phase cells was attributed to the heat-mediated increase in the number of slowly rejoined dsb and partly also to the enhanced fraction of apoptotically dying cells when compared to G1-phase cells. PMID- 11057742 TI - Recombination between homologous chromosomes does not play a dominant role in the formation of radiation-induced chromosomal aberrations. AB - PURPOSE: In mammalian cells, the relevance of homologous recombination in radiation-induced double-strand break (DSB) repair is not yet well understood. In the present work, the role of recombination between homologous chromosomes and homology-directed repair of DSB were studied, using X-ray-induced chromosomal aberrations as an end-point. MATERIALS AND METHODS: Human-hamster hybrid cells containing one or two copies of human chromosome 8 were used. If recombination between homologous chromosomes plays a dominant role in DSB repair, it is expected that X-irradiation of cells with two copies of chromosome 8 would result in a lower frequency of aberrations involving this chromosome compared with cells with only one copy of chromosome 8. The aberrations involving human chromosome 8 were detected by fluorescence in situ hybridization (FISH). Furthermore, a comparison between the hamster cell line XR-C1 (defective in non-homologous repair), CHO-9 (the wild-type cells) and the cell line XR-C1#8 (in which the defect of XR-C1 is complemented by human chromosome 8) was made to determine, indirectly, the involvement of homology-directed recombination in DSB repair. RESULTS: The observed frequencies of aberrations per human chromosome 8 were not significantly different between cells containing one or two copies of this chromosome. The frequency of chromatid-type aberrations was doubled in XR-C1 cells compared with CHO-9 and XR-C1#8 cells. CONCLUSIONS: In hamster cells, recombination between homologous chromosomes appears not to have a major role in the formation of radiation-induced chromosomal aberrations, while nonhomologous repair seems to be important in both the G and G2 phases of the cell cycle. PMID- 11057743 TI - Effect of changing the weekly dose intensity of fractionated irradiation on local control of two human squamous cell carcinomas in nude mice. AB - PURPOSE: To compare the effect of fractionated irradiation with increasing, constant or decreasing weekly dose intensity on local tumour control. MATERIALS AND METHODS: Human squamous cell carcinomas, FaDu and GL, were grown in nude mice. Thirty fractions were applied under ambient conditions with increasing, constant or decreasing weekly dose intensity within a constant overall treatment time of 6 weeks. Dose intensity was changed every 2 weeks. Irradiations were terminated in some groups of animals after 20 fractions in 4 weeks. Endpoint was the tumour control dose 50% (TCD50) at day 120 (FaDu) or day 180 (GL) after end of treatment. RESULTS: In FaDu tumours the TCD50 value of 60 Gy (95% CI 56; 63) for fractionated irradiation with decreasing dose intensity, i.e. high initial doses, was slightly but significantly lower than the TCD50, of 68 (60; 81) after low initial doses (p=0.03). The TCD50 value of 62 Gy (57; 68) after constant doses was intermediate (constant vs increasing p =0.30; constant vs decreasing p=0.15). The higher efficacy of high initial doses in FaDu tumours was explained by local control occurring already during the course of irradiation. In GL tumours the TCD50 values were 52 Gy (43; 62) after high initial dose intensity, 50 Gy (43; 66) after constant doses, and 55 Gy (42; 89) after low initial dose intensity. These values were not statistically different (p-values 0.20-0.75). CONCLUSIONS: The data support the view that initial dose concentration during fractionated irradiation does not enhance radioresistance of FaDu and GL tumours, for instance by an earlier onset of clonogen repopulation. PMID- 11057744 TI - Gold microspheres: a selective technique for producing biologically effective dose enhancement. AB - PURPOSE: To investigate dose enhancement and radiosensitization associated with electrons produced and scattered from gold particles suspended in cells in vitro and with tumour cells growing in vivo irradiated with low-energy photons. MATERIALS AND METHODS: CHO-K1, EMT-6 and DU-145 cells were irradiated with kilovoltage X-ray and Cs-137 beams in slowly stirred suspensions in the presence of various concentrations of gold particles ( 1.5-3.0 microm); cell survival was measured by clonogenic assay. Gold particles were injected directly into EMT-6 tumours growing in scid mice prior to their irradiation. Tumour cell killing was assayed by an in vivo-in vitro technique. RESULTS: Dose enhancement was confirmed by both Fricke dosimetry and cell killing for 100, 140, 200 and 240 kVp X-rays, but not for Cs-137 gamma-rays. For the chemical dosimeter, a dose enhancement (DMF) of 1.42 was measured for 1% gold particle solutions irradiated with 200 kVp X-rays. When rodent and human cells were irradiated in the presence of 1% gold particles, DMF values at the 10% survival level ranged from 1.36 to 1.54, with an overall average value of 1.43. Preliminary attempts to deliver these gold particles to tumour cells in vivo by intra-tumour injection resulted in modest radiosensitization but extremely heterogeneous distribution. CONCLUSIONS: An increased biologically effective dose can be produced by gold microspheres suspended in cell culture or distributed in tumour tissue exposed to kilovoltage photon beams. With the increasing use of interstitial brachytherapy with isotopes that produce low-energy photons, high-Z particles might find a role for significantly improving the therapeutic ratio. PMID- 11057745 TI - A comparison of the degree of curvature in the cancer incidence dose-response in Japanese atomic bomb survivors with that in chromosome aberrations measured in vitro. AB - PURPOSES: To compare the degree of curvature in the dose-response for chromosome aberrations and for radiation-induced cancer. MATERIALS AND METHODS: Comparison of the ratio of the quadratic and linear coefficients (in dose) in Japanese atomic bomb survivor cancer incidence data, based on follow-up to 1987 and taking account of random errors in DS86 dose estimates with the same ratio in four datasets of chromosome aberrations in peripheral blood lymphocytes measured in vitro and exposed to 60Co gamma radiation. RESULTS: There are no statistically significant differences between the four in vitro datasets in the ratio of the quadratic to the linear coefficients for dicentrics or chromosome translocations, nor are there indications of differences between this ratio for dicentrics and that for complete chromosome translocations (p > 0.1 in all cases). If the 0-4 Gy dose range is used in the Japanese atomic bomb survivor data, the ratio of the quadratic to the linear coefficients for all solid cancers is 0.06 -1 (95% CI 0.22, 0.67) and so is not significantly different from 0; this ratio is statistically highly inconsistent (p<0.0001) with the analogous ratio estimated for the in vitro chromosome aberration data (4.20 Sv-1; 95% CI 3.06, 6.51). By contrast, there are no statistically significant differences (p=0.42) between the ratio of the quadratic to the linear coefficients for leukaemia incidence in the Japanese cohort, 1.81 Sv-1 (95% CI 0.21, > 1,000), with that for chromosome aberrations in vitro. These results are not markedly changed if the 0-2 Gy dose range is used in the Japanese atomic bomb survivor data. CONCLUSIONS: It is unlikely that in vitro chromosome aberrations could be a correlate for the initiating radiogenic lesions leading to radiation-induced solid cancers. However, taken together with certain other biological information, it may not be unreasonable on this basis to use in vitro chromosome aberrations in peripheral blood lymphocytes as a correlate of the radiogenic lesions leading to radiation induced leukaemia. PMID- 11057746 TI - Radiation effects in lymphocytes of children living in a Chernobyl contaminated region of Belarus. AB - PURPOSE: To investigate cytogenetic and mutational effects in lymphocytes from individuals chronically exposed to radiation from the Chernobyl catastrophe. MATERIALS AND METHODS: Nine years after the Chernobyl accident (1986), peripheral blood lymphocytes from 20 Kalinkovichi children (age 10-15) and 10 Minsk children (age 10-17) were analysed for genetic damage by several assays. Radiation damage in exposed children was investigated in descendants of progenitor cells that were irradiated during a short period immediately after the accident. In the time-span between the accident and blood sampling the cells were also irradiated chronically by internal radiation originating from ingested radionuclides and, to a smaller extent, by external radiation from radionuclides. The parameters measured in whole blood smears were the frequency of micronucleated mononucleated lymphocytes and binucleated lymphocytes with nucleoplasmic bridges and associated micronuclei. Cultures of cytokinesis-blocked lymphocytes were used to analyse mononuclear and binuclear cells for the presence of micronuclei, also cell killing effects. A colony assay was used to study induction of recessive mutations in the HPRT gene. RESULTS: The analysis of whole-blood smears indicated a doubling of the frequency of micronuclei per 100 mononuclear lymphocytes in exposed children compared with unirradiated children. Small numbers of binucleated lymphocytes with nucleoplasmic bridges and associated micronuclei were found in blood smears from exposed children. Analysis of cytokinesis-blocked cultures indicated in mononuclear cells of exposed children a statistically significant increase in the frequency of micronuclei. When the same parameters were studied in binucleated cells there was no difference between exposed and unexposed children. Results of the dye-exclusion assay showed a four-fold increase in the percentage of dead cells between exposed and unexposed children. There was no evidence for induction of HPRT mutations in exposed children. CONCLUSION: These results indicate that the frequently advocated procedure of simply analysing micronuclei in cytokinesis-blocked binucleated lymphocytes can result in an underestimate of genetic damage induced by radiation accidents. Biodosimetric studies should therefore employ a battery of assays for the detection of several types of genetic damage in different generations of lymphocytes. PMID- 11057747 TI - Cleavage of ATM during radiation-induced apoptosis: caspase-3-like apoptotic protease as a candidate. AB - PURPOSE: To study the relationship of ionizing radiation-induced apoptosis with the integrity of ATM (mutated in ataxia telangiectasia) that has a critical role in DNA damage sensing and repair, cell-cycle checkpoint controls and maintenance of genomic stability. MATERIALS AND METHODS: U937 cells were treated with gamma radiation. Sub-G1 DNA content, DNA fragmentation, cleavage of PARP, active caspase-3, cleavage of ATM in vivo and in vitro were measured by flow cytometry, agarose gel electrophoresis, cleaving of colorimetric caspase-3 substrate and Western blotting. RESULTS: ATM is specifically cleaved in cells during the induction of apoptosis by ionizing radiation exposure. The time-course of cleavage coincided with the appearance of cells with a sub-G1 DNA content and activation of caspase-3. ATM was cleaved with similar kinetics as PARP and DEVD FMK could abolish the cleavage. In vitro studies showed that ATM was cleaved by caspase-3 or related subfamily members at a DIVD/G site. CONCLUSION: ATM belongs to a group of repair proteins, including PARP, DNA-PK and HsRad51, which are specifically cleaved during apoptosis. These findings support the idea that repair mechanisms need to be inactivated for apoptosis to proceed efficiently. PMID- 11057748 TI - Effect of irradiation at the early fetal stage on adult brain function in the mouse: locomotor activity. AB - PURPOSE: To study long-term changes in the adult locomotor activity of mice after exposure to gamma radiation at the early fetal stage of development. MATERIALS AND METHODS: Pregnant Swiss albino mice were exposed locally on the abdominal area to a single dose of 0.25-1.5 Gy of 60Co gamma radiation at the dose rate of 1 Gy/min. When the F1 offspring were 6 months old, their locomotor and exploratory behaviour was assessed by the open-field and dark/bright arena tests. Animals were again subjected to the dark/bright arena test at 12 and 18 months of age in order to study the persistence of the effects. RESULTS: Irradiation produced a noticeable disturbance in the normal behaviour pattern of the mice. There was a significant dose-dependent decrease in the open-field activity of 6 month-old mice. In the dark/bright arena test, the time spent and lines crossed in the dark area showed a significant decrease, while their activities in the brightly lit area increased significantly, indicating a reduced aversion to bright light. These effects were evident even at a dose of 0.3 Gy and increased linearly with dose. The significant behavioural changes persisted at 12 months, but at 18 months the difference in the time spent and lines crossed in the dark and bright areas were not significantly different from sham-irradiated control values below 0.5 Gy. CONCLUSIONS: These results show that day 14 of gestation in Swiss albino mice is a time of high risk for inducing long-term changes in the adult locomotor function by gamma-radiation doses below 1 Gy. Using a range of radiation doses and different observation times we have demonstrated that the effect increases linearly with dose, but there appears to be a threshold of 0.3 0.5 Gy for producing significant persistent changes in the adult ambulatory activity. PMID- 11057749 TI - Effects of ionizing radiation on oocytes of prepubertally irradiated rats. AB - PURPOSE: To characterize the effects of ionizing radiation on the immature oocytes of prepubertal rats aged 16-20 days old. MATERIALS AND METHODS: The abdomen of Sprague Dawley (SD) female rats was exposed to 0, 1 and 2 Gy of X rays. The effects were analysed when the animals became adults (80 days old) using as parameters the number of implantation sites, embryonic loss, dominant lethality and the synaptonemal complex (SC) analysis of F1 female foetuses. RESULTS: A statistically significant decrease in the fertility of the 2 Gy irradiated rats was observed. However, the SC analysis of F1 female foetuses showed no cytogenetic damage that could be attributed to radiation. CONCLUSIONS: A reproductive effect was demonstrated but no inherited genetic damage was detected by SC analysis in female foetuses of irradiated mothers. PMID- 11057750 TI - Mobilization and detoxification of polonium-210 in rats by 2,3-dimercaptosuccinic acid and its derivatives. AB - PURPOSE: To reduce retention and toxicity of the alpha particle emitter polonium 210 in rats by newly developed chelating agents. MATERIALS AND METHODS: Repeated subcutaneous chelation was conducted after intravenous injection of 210Po nitrate. For reduction of 210Po retention the treatment with vicinal dithiols meso-and rac-2,3-dimercaptosuccinic acid (DMSA), mono-i-amylmeso-2,3-dimercapto succinate (Mi-ADMS) and mono-N-(i-butyl)-meso-2,3-dimercapto succinamide (Mi BDMA) were used. For the reduction of toxic effects of 210Po, treatment effectiveness of Mi-BDMA was compared with that of N,N'-di(2 hydroxyethyl)ethylenediamine-N,N'-biscarbodithioate (HOEtTTC, reference compound). RESULTS: Treatment with meso-DMSA and rac-DMSA altered the main excretion route of 210Po, reduced its contents in the liver but increased its deposition in the kidneys. Treatment with Mi-ADMS or Mi-BDMA increased total excretion of 210Po, mainly via the faeces. Only Mi-BDMA decreased 210Po levels in the kidneys. The effectiveness of all chelators decreased with delay in the start of treatment. In a survival study, the lives of rats treated early with Mi-BDMA or delayed with HOEtTTC were prolonged three-fold when compared with rats receiving a lethal amount of 210Po only. CONCLUSIONS: Of the vicinal dithiols examined, Mi-BDMA was the best mobilizing chelating agent for 210Po and it reduced 210Po toxicity when the treatment started immediately. However, the detoxification efficacy of the immediate treatment with HOEtTTC, observed in our previous study, was superior to that of the present result with Mi-BDMA. PMID- 11057751 TI - Contamination and decontamination of rat and human skin with plutonium and uranium, studied with a Franz's chamber. AB - PURPOSE: Our work offers a new method of assessing human skin radiocontamination and of appraising its treatment. This in vitro technique stems from methods used in skin pharmacology. MATERIALS AND METHODS: Franz's diffusion chambers, which help maintain the physiological condition of a skin biopsy, are used to study how 233U and 239Pu, added to a 0.1 N solution of nitric acid, could enter skin. The efficiencies of two different decontaminating agents (Na3Ca)DTPA (25%) and EHBP (0.5 M) are compared. These studies were made on human skin samples, recovered after plastic surgery. Parallel experiments were carried out on hairless rat skin biopsies and on the skin of live hairless rats. RESULTS: Results in vivo and in vitro on the rat were not significantly different which validates the in vitro technique. In human beings, most of the radioactivity was found in the epidermis, with 2-4% found at the level of the dermis. By means of autoradiography we were able to identify that this radioactivity was concentrated in and around hair and sebaceous glands. Local treatments by EHBP seemed more efficient than those by DTPA in decorporating uranium and plutonium but the complexes radionuclide-EHBP seemed to diffuse through the skin more than the radionuclide-DTPA complexes if the skin was not rinsed after application of the chelating agent. CONCLUSION: This new in vitro technique using human skin has been validated for radiotoxicology. It can be used to quantify the diffusion of radionuclides through the various skin layers and to assess the efficiency of decontaminating agents. PMID- 11057752 TI - On the dose-effect relationship of true simple chromosomal rearrangements. Comments on the paper: simple chromosome exchanges are not linear with dose. PMID- 11057753 TI - Distinct accumbal subareas are involved in place conditioning of amphetamine and cocaine. AB - In considering the heterogeneous function of the nucleus accumbens (NAC), the present work evaluated the conditioned place preference (CPP) after local infusion of d-amphetamine (AMP; 10, 15 microg/side) or cocaine (COC; 50, 100 microg/side) into two subareas of NAC, core and shell. A regular two-compartment CPP apparatus was used to test the place conditioning effects after 6 pairings of drug in one compartment and 6 pairings of vehicle in the other one. Significant CPP was observed with either AMP infused in the core area or COC infused into the shell area. Neither AMP in shell nor COC in core significantly produced CPP. These results indicate important differences between two neural substrates within NAC for the rewarding effects of AMP and COC on the CPP task. PMID- 11057754 TI - Effect of cAMP on protein binding activities of three elements in upstream promoter of human CYP11A1 gene. AB - CYP11A1 gene encodes the P450scc enzyme. which catalyzes the first step of steroidogenesis under the modulation of cAMP signal. To understand the signaling pathway of cAMP, we analyzed three elements in upstream cAMP-responsive sequences of CYP11A1 gene. The sequence of -1617/-1609 fragment was identical to the conserved SF1 binding site (SF1/Ad4BP). Both sequences of -1637/-1626 and -1559/ 1545 fragments are highly homologous to consensus TRE or CRE. The cAMP-treated adrenal Y1 nuclear extracts formed significantly higher intensities of complexes with these three elements than untreated Y1 nuclear extracts. The EMSA demonstrated potential binding of SF1 protein to -1617/-1609, and CREB like protein and AP1 family to -1637/-1626 and -1559/-1545. This is the first report to describe the enhancement of cAMP on protein binding activities of three elements of CYP11A1 promoter to SF1/Ad4BP or CREB/AP1-like proteins. PMID- 11057755 TI - Activity of phenolsulfotransferases in the human gastrointestinal tract. AB - Sulfate conjugation by sulfotransferase enzymes is an important pathway for the detoxication of xenobiotics and endogenous compounds. The large surface area of the gastrointestinal tract exposes the body to a range of potential toxins, and hence local metabolism is likely to be important. The ability of different regions of the gut to sulfate micromolar concentrations of simple phenols and catecholamines has been determined throughout the gut using 4-nitrophenol and dopamine as standard substrates. The pattern of sulfation of both compounds was similar, with activity highest in the small bowel >right colon >left colon >rectum >stomach >esophagus. High concentrations of sulfotransferases in the reservoir areas of the right and left colon indicate possible importance in detoxication by sulfation and also perhaps in activating mutagens in the same areas. Nutritional factors, such as a high-fat diet may, however, alter sulfotransferase activity. PMID- 11057756 TI - Inhibition of malignant trophoblastic cell proliferation in vitro and in vivo by melatonin. AB - Melatonin inhibited thymidine incorporation into human choriocarcinoma JEG-3 cells at physiological and pharmacological concentrations in the present study. Gene expression of MT2 receptor, but not that of mt1 receptor, was detected in JEG-3 cells by reverse transcription-polymerase chain reaction (RT-PCR). The gene expression profile of the two human melatonin receptor subtypes in JEG-3 cells was identical to that previously reported for JAr cells, whose proliferation had also been shown to be similarly inhibited by physiological and pharmacological concentrations of melatonin. In contrast, melatonin had no effect on thymidine incorporation into 3A-Sub-E cells (a transformed trophoblast cell line), in which gene expression of both receptor subtypes could not be detected. The data suggest that in human placental trophoblasts, a correlation may exist between MT2 receptor gene expression and the direct anti-proliferative action of melatonin. Although melatonin has been reported to induce G1/S delay in cell cycle progression of JAr cells, no significant changes in the percentages of JEG-3 cells in different cell cycle phases upon melatonin treatment was recorded by flow cytometric analysis. This indicates that G1/S transition delay is probably not an important cellular mechanism in the direct anti-proliferative action of melatonin on human JEG-3 cells in vitro. Furthermore, melatonin inhibited the growth of both JAr and JEG-3 xenograft tumors in athymic nude mice, and prolonged the survival of those animals that developed choriocarcinoma. While the number of apoptotic tumor cells was not increased by melatonin, the pineal hormone induced significant decreases in the numbers of JAr and JEG-3 cells expressing proliferating cell nuclear antigen (PCNA) and cyclin A in the tumors. Taking into account both the in vitro and in vivo findings, it is likely that the inhibitory effect of melatonin on choriocarcinoma JAr and JEG-3 cell proliferation in vivo is largely a direct action of the hormone on the tumor cells. PMID- 11057757 TI - Further studies on 20beta-hydroxysteroid dehydrogenase with carbonyl reductase like activity present in liver microsomes of male rats. AB - Further characterizations of 20beta-hydroxysteroid dehydrogenase (20beta-HSD) present in liver microsomes of male rats were examined. A significant relationship was observed between 20beta-HSD and acetohexamide reductase (AHR) activities in liver microsomes of male rats. The hepatic microsomal 20beta-HSD and AHR preferentially required NADPH as a cofactor. When NADPH was replaced by NADH, NADP or NAD at the same concentration, these reductase activities were little detected. The hepatic microsomal 20beta-HSD and AHR activities in streptozotocin-induced diabetic rats were much lower than those in the corresponding controls. The hepatic microsomal 20beta-HSD and AHR activities appeared as one main peak, respectively, on DEAE-Sephacel column chromatography, and the peak of 20beta-HSD activity was in good agreement with that of AHR activity. Based on these results, we conclude that 20beta-HSD present in liver microsomes of male rats functions as AHR, and exhibits a carbonyl reductase-like activity. PMID- 11057758 TI - Effects of hypoxia on cholesterol metabolism in human monocyte-derived macrophages. AB - We assessed the metabolism of low density lipoprotein (LDL) of human monocyte derived macrophages under hypoxia. The specific binding and association of 125I labeled LDL (125I-LDL) were not changed under hypoxia compared to normoxia. However, the degradation of 125I-LDL under hypoxia decreased to 60%. The rate of cholesterol esterification under hypoxia was 2-fold greater on incubation with LDL or 25-hydroxycholesterol. The cellular cholesteryl ester content was also greater under hypoxia on incubation with LDL. Secretion of apolipoprotein E into the medium was not altered under hypoxia, suggesting that apolipoprotein E independent cholesterol efflux may be reduced under hypoxia. Thus, hypoxia affects the intracellular metabolism of LDL, stimulates cholesterol esterification, and enhances cholesteryl ester accumulation in macrophages. Hypoxia is one of the important factors modifying the cellular lipid metabolism in arterial wall. PMID- 11057759 TI - Troglitazone inhibits the expression of inducible nitric oxide synthase in adipocytes in vitro and in vivo study in 3T3-L1 cells and Otsuka Long-Evans Tokushima Fatty rats. AB - The aim of this study was to determine the mechanism of troglitazone action on nitric oxide (NO) production via inducible NO synthase (iNOS) in adipocytes in vitro and in vivo. The treatment of 3T3-L1 adipocytes with the combination of lipopolysaccharide (LPS), tumor necrosis factor-alpha and interferon-gamma synergistically induced de novo iNOS expression leading to enhanced NO production. The NO production was inhibited by co-treatment with aminoguanidine or N-nitro-L-arginine methylester hydrochloride. Troglitazone inhibited the NO production in a dose dependent manner by the suppression of iNOS expression. In the 24 week-old Otsuka Long-Evans Tokushima Fatty (OLETF) rats, the mean weight and the blood glucose were 21% and 30%, respectively, higher than in their lean counterparts. The serum nitrite concentration was increased after injection of LPS (4 mg/kg, i.p.), more markedly in OLETF rats than in the lean rats. The epididymal fats from LPS-injected groups, but not the ones from the non-injected groups, expressed mRNA and protein of iNOS. Troglitazone pre-treatment blocked the LPS-induced expression of iNOS in adipose tissue and the increase in serum nitrite concentration. These results suggest that troglitazone inhibits the cytokine-induced NO production in adipocytes by blocking iNOS expression both in vitro and in vivo. PMID- 11057760 TI - Growth enhancement of fowls by dietary administration of recombinant yeast cultures containing enriched growth hormone. AB - In present study the methylotrophic yeast, Pichia pastoris, was used to express a recombinant growth hormone (rGH) gene of swine. A synthetic secretion cassette was constructed using the promoter of the alcohol oxidase1 gene (AOX1), and a alpha-factor signal peptide. After electroporatic transformation and zeocin selection, several clones exhibited high levels of rGH protein expression constituting more than 20% of total yeast protein. Over 95% of rGH was shown to be export into the culture supernatant. Yeast transformant containing the highest recombinant growth hormone level (rGH yeast) and native GS115 Pichia pastoris (non-rGH yeast, as a control) were separately cultured, harvested and adsorbed by wheat bran. Yeast cultures of four dosages (0.05, 0.1, 0.2, and 0.4%) were mixed respectively with chick basal diet and fed to simulated country chickens for 9 weeks. The results showed that, when compared to control chicks, the percentage of body weight gain was improved significantly (P<0.05) in chicks fed with diets containing 0.1 or 0.2% rGH-rich yeast culture at brooding stage, and in chicks fed with 0.4% rGH-rich yeast culture at growing stage. The average weight gain in rGH yeast treated groups for the full-term (0 to 63d) and short term (43 to 63d) of growth were 10.6 and 9.4%, respectively, better than the non-rGH yeast control group. These experimental data suggest that the use of rGH-containing yeast as a supplement in fed provided an alternative approach for growth improvement in simulated country chickens. PMID- 11057761 TI - Inhibition of P-glycoprotein expression and reversal of drug resistance of human hepatoma HepG2 cells by multidrug resistance gene (mdr1) antisense RNA. AB - The development of multiple drug resistance in tumor cells is a significant problem in cancer therapy. In human, one of the reasons causing the resistance is due to the overexpression of the mdr1 gene product, P-glycoprotein. In our study, we had developed multiple drug resistant HepG2 cell line (HepG2/DR). To reverse the resistance, HepG2-DR cells were treated with antisense RNA against mdr1 gene. Total RNA and protein were extracted from the transfected cells. Northern analysis showed that mRNA level of mdr1 was decreased whereas a reduction in P glycoprotein was detected by Western blot. By using flow cytometry, the ability of intracellular doxorubicin retention increased and drug efflux decreased in the treated cells. The result also showed that the cellular sensitivity to doxorubicin, vincristine and methotrexate measured in IC50 increased 83.3% 84.6% and 50% respectively. All these findings suggested that the expression of p glycoprotein was successfully inhibited by antisense RNA and the drug resistance was reduced. PMID- 11057762 TI - Effect of aging on the modulation of macrophage functions by neuropeptides. AB - The existence of a functional connection between the nervous and the immune system is supported by increasing recent evidence. In previous work we have shown that peptides from the nervous system, such as gastrin-releasing peptide (GRP), neuropeptide Y (NPY) and sulfated cholecystokinin octapeptide (CCK-8s), have modulatory effects on the immune functions in adult animals. Since the immunodepression found in aging organisms may be related to changes in the neuroimmune network, the aim of the present work was to study the changes with aging in the effect of CCK-8s, GRP and NPY on peritoneal macrophage functions (adherence to tissues, mobility, ingestion of foreign particles and superoxide anion production) from BALB/c mice of three different ages: adult (24+/-2 weeks old), mature (50+/-2 weeks old) and old (72+/-2 weeks old). The results show that the increase in adherence capacity produced by neuropeptides in cells from adult and mature animals disappears in old mice. The stimulatory effect of GRP and NPY on mobility, ingestion and superoxide production in macrophages from adult mice disappears (GRP) or changes to inhibition (NPY) in cells from old animals. The decrease of these functions caused by CCK-8s in adult or mature animals continues in old mice. These data suggest that the modulation by neuropeptides of the macrophage function changes with the age of animals. PMID- 11057763 TI - Anti-analgesic and anti-amnesic effect of complement C3a. AB - In the present study, we found that complement C3a exerted central effects after intracerebroventricular administration in mice. At doses of 3 and 10 pmol/mouse, the peptide showed an antagonistic effect on analgesia induced by morphine and U 50488H, known to be mu- and kappa-opioid receptor agonists, respectively. Moreover, complement C3a improved scopolamine- and ischemia-induced amnesia at a dose of 10 pmol/mouse. Anti-analgesia was not observed by C3a des-Arg at 10 pmol/mouse. The present findings suggest that complement C3a may act as a peptide with anti-opioid activity in the central nervous system. PMID- 11057764 TI - Urogenital injuries in childhood: a strong association of bladder trauma to bowel injuries. AB - We analysed the inter-relationships of the cause and the type of trauma, the presence of pelvic fracture, the associated intraabdominal organ injuries, and the morbidity and mortality rates in 154 patients presenting and being treated for UGT between 1983 and 1997. The cause of injury was blunt in 77% of cases and penetrating in 13%. The most frequently injured organs were kidney followed by urethra and bladder. Bowels, liver and spleen were the most frequently associated injured organs. Moreover, bladder injuries were strongly associated with bowel injuries (p < 0.0001). Hemodynamically normal 49 children with minor or major kidney injuries were managed conservatively. Hemodynamically non-stable 11 patients were explored. The majority of urogenital injuries can be managed conservatively even when associated with intraabdominal organ injuries. Solid genitourinary organ injuries may accompany more frequently to intraperitoneal solid organ injury. Whereas, non-solid genitourinary organ injuries may more frequently associated with injuries of intraperitoneal hollow viscus. PMID- 11057765 TI - Posterior retroperitoneoscopic adrenalectomy for unilateral nodular hyperplasia in a hemodialaysis patient. AB - We report a case of posterior retroperitoneoscopic adrenalectomy for unilateral nodular hyperplasia in a patient with chronic renal failure on hemodialysis. Posterior retroperitoneoscopic adrenalectomy appears to be a safe and valuable alternative for the treatment of the adrenal tumors in hemodialysis patients with easy tissue fragility and bleeding tendency. PMID- 11057766 TI - Primary megaureter detected by prenatal ultrasonography: conservative management and prolonged follow-up. AB - With the widespread use of obstetric echography the incidence of fetal hydronephrosis has been reported more frequently. Consequently, many uropathies have been detected in asymptomatic neonates. The authors report their experience with prenatally detected primary non-refluxing megaureter. Newborns with fetal hydronephrosis were investigated by ultrasonography and micturating cystourethrogram after the beginning of chemoprophylaxis. If primary megaureter was identified, after 1 month the children underwent 99tm-DMSA, diuretic 99tm DTPA, and intravenous urography. Eight infants with primary megaureter (bilateral in 3 cases) were identified, for a total of 11 renal units for study. All children were submitted to non-operative management. We performed ultrasonography and diuretic 99tm-DTPA during follow-up, which lasted on average 75 months. The mean cross-sectional diameter of the dilated ureter was 13.6 mm during neonatal period, and reached 8.4 mm at the end of follow-up. The renal function and the diuretic renogram remained stable throughout follow-up. Two neonates presented transitory hypertension. Our results support the notion that conservative management is safe for primary megaureter detected in asymptomatic neonates, with most cases showing spontaneous regression during a prolonged follow-up. PMID- 11057767 TI - Kinetic versus thermodynamic factors in calcium renal lithiasis. AB - Calcium renal lithiasis formation depends on the balance between thermodynamic (supersaturation) and kinetic (inhibitors, nucleants) factors. In this paper, the importance of both groups was evaluated using (a) the complete urine analysis data obtained from 32 healthy volunteers and 141 active stone-formers, and (b) a comprehensive computer model to calculate the supersaturation values of calcium oxalate monohydrate, hydroxyapatite and brushite in each urine sample. The results of this evaluation were used to assess the possible effectiveness of a given pharmacological treatment. PMID- 11057768 TI - Trapping of the double-J stent in the urinary tract eight years after extracorporeal shock wave lithotripsy. AB - We report a case of the 'forgotten' D-J stent in the urinary tract for eight years. In spite of years D-J stent spent in the urinary tract, and the great burden of the new-formed stones, residual renal function still exist. One year after treatment patient is stone free, infection free, with the mild renal failure. PMID- 11057769 TI - Is visualising ureter before pyeloplasty necessary in adult patients? AB - In this study, we aimed to detect whether or not visualising ureter and ureteropelvic junction (UPJ) preoperatively is necessary in adult patients who have primer UPJ obstruction. Between January 1995 to June 1999, 46 renal units in 45 patients with primer UPJ obstruction were evaluated. The patients were separated into 2 groups. In group 1, intravenous pyelography (IVP) and renal scintigraphy were performed to 17 renal units preoperatively. In group 2, in addition to these methods, either retrograde pyelography (RGP) or antegrade pyelography (AGP) were performed to 29 renal units. Renal/bladder sonogram was used in patients with poor renal function in IVP or in renal scintigraphy. All the operations were performed through a flank incision. In group 2, additional information was gained for 8 (27.5%) of the renal units preoperatively. No additional information for this group found intraoperatively. In group 1, we found additional information in 4 (23.53%) of the units intraoperatively. All the pathologies in both groups were corrected intraoperatively. Double-J (D-J) stent was used in 6 (35.29%) of the units in group 1 and 8 (27.58%) of the units in group 2 intraoperatively (p > 0.05). In group 2, 4 (13.79%) preoperative complications were seen due to RGP and they were treated either medically or conservatively. In the early postoperative period, a complication observed in 1 (5.88%) of the patients in group 1 and 1 of the patients in group 2 (3.44%) (p > 0.05). The first patient was treated with inserting D-J and the latter one was treated conservatively. In the 3rd postoperative month, success rate was found to be 94.11% in group 1 and 96.55% in group 2 (p > 0.05). Additional pathologies in adult patients with primer UPJ obstruction can be corrected intraoperatively through a flank incision. Therefore, imaging of ureter and UPJ may not be necessary in these patients. PMID- 11057770 TI - Laparoscopic removal of a gigantic non-functioning kidney via retroperitoneal access. AB - Hugely dilated kidneys can sometimes present as abdominal masses. These kidneys are invariably non-functioning and are managed by nephrectomy. We describe a case of massive kidney containing 12.5 litres on fluid which was managed by retroperitoneoscopic nephrectomy. The patient was a 24-year-old male who presented with a huge abdominal mass, anorexia and weight loss. Laparoscopic surgery for such a large kidney has not been previously reported. We discuss salient features of the procedure and elaborate on the modifications required in the case of significantly enlarged kidneys. PMID- 11057771 TI - Isolated renal and retroperitoneal hydatid cysts. AB - Four patients (3 male, 1 female) with isolated renal and 1 female patient with isolated retroperitoneal cysts were reviewed. The mean age of the patients was 46 (25-64). The most common presenting symptom was pain. Two cases were discovered incidentally by the observance of renal calcification on abdominal x-ray. Indirect hemagglutination test was positive in all cases but eosinophilia was present only in 1 (20%) case. Nephrectomy was performed to 1 patient who presented with hydaturia and had a large communicating cyst involving most of the kidney. Total cystectomy was performed in other renal cysts. Total cystectomy with wide excision of the involved muscle was performed to the retroperitoneal hydatid cyst. Patients were followed by an average of 23.8 (9-50) months with indirect hemagglutination test and USG. No evidence for recurrence was found up to date. PMID- 11057772 TI - Simplification of laparoscopic extraperitoneal colposuspension: results of two port technique. AB - PURPOSE: The aims of the present prospective study were to apply a new simplification for Laparoscopic Burch Colposuspension and to assess the postoperative results of this treatment modality for stress urinary incontinence. MATERIALS AND METHODS: Twenty-four patients underwent simplified laparoscopic procedure via two ports, utilising endoscopic tacker and two stripes of prolene mesh. Patients were evaluated for operative time, duration of urethral catheter, length of hospital stay, complications and continence status. RESULTS: The technique was successful in 21 patients after 24 months follow-up. Average operative time was 39 minutes. The lengths of urethral catheterisation and hospital stay were 22 and 45 hours, respectively. One operative complication and three postoperative recurrences were recorded. CONCLUSION: Laparoscopic bladder neck suspension continues to develop as the instruments and techniques are improved. In carefully evaluated patients, it provides safe and minimally invasive procedure. We report the results of a new technique to simplify and speed up a laparoscopic extraperitoneal colposuspension. PMID- 11057773 TI - The best management of superficial bladder tumours: comparing TUR alone versus TUR combined with intravesical chemotherapy modalities? AB - To compare retrospectively the recurrence rates of TUR alone versus different intravesical chemotherapy modalities in superficial bladder cancer cases, 187 patients with stage Ta and T1 bladder tumours were treated with transurethral resection followed by adjuvant intravesical chemotherapy with mitomycin, BCG or epirubicin or by transurethral resection alone. All patients in this study had historically proven transurethrally resectable primary, category Ta and T1 transitional cell carcinoma (TCC) of the bladder. Group I included transurethral resection alone, and the other groups included intravesical mitomycin-C (Group II), BCG (Group III) and epirubicin (Group IV) therapies after transurethral resection. 146 male and 41 female patients (78% male and 22% female patients) in this study were diagnosed as primary TCC bladder tumours. Only 52 of them were stage Ta and 135 of them were stage T1 bladder tumours. Examining the histological grade of the bladder tumours, 88 (47%) of the patients had grade I, 53 (28%) had grade IIa, 30 (16%) had grade IIb and remaining 16 (9%) had grade III bladder cancers. The recurrence rates were 25% for Group I, 23.8% for Group II, 26.2% for Group III and 22.7% for Group IV. These values were given with disregarding the grade and volume of the bladder tumours. For solitary, less than 3 cm low grade tumours (grade I, IIa) recurrence rates were 16% for Group I, 15.4% for Group II, 17.8% for Group III, 17.2% for Group IV (p > 0.05). As a result of this retrospective study, for patients with low grade, stage Ta and T1 tumours TUR alone may be the best treatment modality. Although intravesical chemotherapy is effective in decreasing short-term incidences of tumour recurrence, it has not decreased long-term incidences of tumour recurrence. The high cost and adverse side effects of intravesical chemotherapy should also be taken into consideration in superficial, single, low grade tumours of bladder. PMID- 11057774 TI - Adjuvant chemotherapy for non-organ confined disease after radical cystectomy. AB - Of 50 patients with pT3b, pT4 and/or pN+ disease after cystectomy, 27 were administered adjuvant four cycles of cisplatin, methotrexate and vinblastine (CMV) and 23 were followed expectantly (no-treatment group). Median follow-up was 14 months in CMV group and 11 months in no-treatment group. Median recurrence free survival was 21 months in CMV group and 17 months in no-treatment group (p = 0.573). Median overall survival was 41+ months in CMV group and 84+ months in no treatment group, respectively (p = 0.501). In our experience, adjuvant chemotherapy after cystectomy seemed not to provide a survival advantage. PMID- 11057775 TI - Concomitant presence of bladder cancer and neurogenic bladder in a patient with HTLV-1 carrier: a case report. AB - We describe a case of an HTLV-1 carrier who developed bladder cancer and neurogenic bladder. HTLV-1 is thought to alter host immune function and to contribute to the development of other malignancies. It is also sometimes reported that urinary symptoms precede pyramidal symptoms in patients with HAM. To our knowledge, concomitant presence of bladder cancer and neurogenic bladder in an HTLV-1 carrier has not been previously reported. PMID- 11057776 TI - Alpha adrenergic blockers in the treatment of benign hyperplasia of the prostate. AB - Benign prostatic hyperplasia (BPH), a common benign tumor in men has been attributed to age and male androgen functions. Of the various management options for treatment of BPH, medical therapy is the first line treatment modality involving either blockade of alpha adrenergic receptors or inhibition of 5-alpha reductase. Amongst these, the alpha-1 blockers are used most frequently. The association of numerous adverse effects with non selective and short acting alpha 1 blockers (like phenoxybenzamine, prazosin and alfuzosin) has led to the development of long acting alpha-1 adrenoceptor blockers (doxazosin, terazosin, tamulosin) which being uroselective significantly reduce the incidence of cardiovascular side effects and increase patient compliance. The review gives a brief account of pharmacological properties and efficacy of alpha adrenergic receptor blockers in the treatment of BPH. PMID- 11057777 TI - Observations on EGFR gene amplification and polymorphism in prostatic diseases. AB - This study was designed to determine the amplification and polymorphism of epidermal growth factor receptor (EGFR) gene in prostatic diseases like benign hypertrophy (BPH) and carcinoma (CaP). The EGFR gene was found to be amplified in grade IV BPH as compared to grade II BPH. Digestion of genomic DNA with MspI and HpaII revealed the presence of a 5kb band following southern blot analysis. This 5kb band was present in all the CaP cases and in one out of three BPH cases. It is possible that such a polymorphism is associated with the type or extent of tumor progression. PMID- 11057778 TI - 'Hot flush', an unpleasant symptom accompanying antiandrogen therapy of prostatic cancer and its treatment by cyproterone acetate. AB - From 1995 to 1997 the authors have assessed 31 patients with histologically verified advanced carcinoma of the prostate (CaP) and the ensuing symptom of 'hot flush'. Patients underwent transurethral resection of the prostate (TURP), bilateral orchiectomy (OE) and combined androgen blockade (CAB) by the administration of non-steroid antiadrogens. The authors present the mechanism of the genesis of the 'hot flush' symptom as well as its subjective manifestations, methods of laboratory monitoring as well as their experience with the treatment of this symptom. 50 mg tablets cyproterone acetate administered twice daily or Androcur depot 300 mg i.m. inj. once in 14 days were the main factors in the treatment of 'hot flushes' which reduced subjective difficulties in 80.6% of the patients studied. PMID- 11057779 TI - Estradiol and nitric oxide in men over 50 years of age. AB - We measured the circulatory levels of nitric oxide and estradiol in men over 50 years with estrogenisation findings. We investigated relation between nitric oxide, which is the principal signal for relaxation of vascular smooth muscle cells, and estradiol levels in serum in men over 50 years with estrogenisation. This study included 14 men with (group 1) and 20 without estrogenisation findings (group 2). Mean nitric oxide and estradiol levels were found to be significantly lower in group 2 than group 1. Group 2 had lower levels sex-hormone binding globin and higher both systolic and diastolic blood pressure values than group 1. There was a significant correlation between nitric oxide and estradiol in esrogenisation group. PMID- 11057780 TI - Comparison of preputial sac and urine cultures in healthy children. AB - The aim of the present study was to examine subpreputial bacteriology and to compare it with the urine cultures of healthy male children. Seventy-two male children were divided into two groups as A and B according to age. In both groups preputial sac and urine cultures were taken simultaneously. Gram (+) enteric cocci were the most common isolated pathogens from the preputial sac in group B. Enterobacter, E. coli and staphylococci species were isolated from the urine cultures of three patients in group B. We could not find any difference between the preputial sac swabs of group A and B patients, but the isolation rate of urine cultures of group A patients was significantly higher than group B (p < 0.05). The findings of the present study support a potential role of the prepuce acting as a reservoir of faecal bacteria in the pathogenesis of UTI in male infants, especially in the first year of life. Improved penile hygiene after the first year of life does not alter the subpreputial bacteriology, but significantly decreases the contamination of urine. PMID- 11057781 TI - Does pleurodesis for pleural effusions give bright ideas about the agents for hydrocele sclerotherapy? AB - OBJECTIVE: Evaluation of the effectiveness of rifampicin and some agents used in the pleurodesis of pleural effusions, such as autologous blood and purified mineral talc. PATIENTS AND METHODS: A total of 56 hydroceles were treated by sclerotherapy, in a random fashion, using purified mineral talc, rifampicin and autologous blood as sclerosant agents. The control group of patients were handled with aspiration only. RESULTS: The cohort of patients in the blood group had a success rate comparable to the control group (p > 0.05). the rifampicin group did better than both control and blood groups (p < 0.05) but not better than the talc group (p < 0.01). Success rate was highest in the talc group of patients who needed no re-sclerotherapy procedures. CONCLUSION: Purified mineral talc was shown to be potentially the best sclerosant for the sclerotherapy of hydroceles and epididymal cysts. PMID- 11057782 TI - Intestinal testis tumor metastasis as a cause of intussusception: a case report. AB - We report on an extraordinary testicular tumour causing intussusception with its intestinal metastases. PMID- 11057783 TI - Leiomyoma of the epididymis. AB - Tumours of the epididymis, both primary and secondary, whether benign or malignant are very rare. We report a case of leiomyoma of the epididymis and discuss the treatment of this tumour. PMID- 11057784 TI - Hypertensive renovasculopathies and the rise of blood pressure with age in Japan and USA. AB - Arterial intimal fibroplasia in renal interlobular arteries but not arteriolar hyalinization was reported to be proportional to the rise of blood pressure with age in the data from all populations examined so far. New findings from Japan offer further insights into the disparities between the two types of renovasculopathy, both of which are called by the same name, 'arteriolosclerosis'. PAS stained paraffin sections were prepared from specimens obtained at autopsy in Tokyo and New Orleans, emphasizing basal subjects, i.e. those with no cause of death known to be related to hypertension. Severities of fibroplastic vasculopathy, in units of intimal thickness as % of outer diameter, and hyaline vasculopathy, in units of affected arterioles per cm2 of tissue sectional area, were measured morphometrically. Blood pressure data were taken from published population surveys. Fibroplastic renovasculopathy was found to provide a proxy for mean blood pressure (MBP) when comparing groups of men and women of various age groups in the USA and Japan. Hyaline renovasculopathy did not reproduce these patterns. Some of these findings confirm similar results from prior studies, and this reproducibility increases confidence that a true biological difference may exist between these populations. PMID- 11057785 TI - Hydronephrosis and renal deterioration in the elderly due to abnormalities of the lower urinary tract and ureterovesical junction. AB - The geriatric population presents a unique challenge to the health care provider. The incidence of common lower urinary tract disorders, such as benign prostatic hypertrophy (BPH), prostate cancer and incontinence increase dramatically with aging. In their more severe forms, these disorders may predispose to hydronephrosis and ultimately to renal deterioration. This review of lower urinary tract and ureterovesical junction (UVJ) physiology and pathophysiology, will focus on: (1) anatomic UVJ obstruction from prostate cancer, or severe bladder hypertrophy, (2) functional obstruction from compression or stretching of the UVJ during bladder distention from urinary retention, and (3) bladder decompensation in the female. We will present a diagnostic and treatment algorithm and discuss future trends in the geriatric population. Clearly, the geriatric health care provider always must consider the lower urinary tract when confronted with acute renal deterioration, because prompt diagnosis and treatment of significant, lower-urinary-tract disease can maximize recovery of renal function. PMID- 11057786 TI - Hemodialysis in elderly patients. PMID- 11057787 TI - The clinical physiology of aging. PMID- 11057788 TI - Mental illness, muscle rigidity in tremors and renal failure. PMID- 11057789 TI - Geriatric nephrology and urology literature. PMID- 11057790 TI - Treatment with exogenous surfactant stimulates endogenous surfactant synthesis in premature infants with respiratory distress syndrome. AB - OBJECTIVE: Treatment of preterm infants with respiratory distress syndrome (RDS) with exogenous surfactant has greatly improved clinical outcome. Some infants require multiple doses, and it has not been studied whether these large amounts of exogenous surfactant disturb endogenous surfactant metabolism in humans. We studied endogenous surfactant metabolism in relation to different amounts of exogenous surfactant, administered as rescue therapy for RDS. DESIGN: Prospective clinical study. SETTING: Neonatal intensive care unit in a university hospital. PATIENTS: A total of 27 preterm infants intubated and mechanically ventilated for respiratory insufficiency. INTERVENTIONS: Infants received a 24-hr infusion with the stable isotope [U-13C]glucose starting 5.3 +/- 0.5 hrs after birth. The 13C incorporation into palmitic acid in surfactant phosphatidylcholine (PC) isolated from serial tracheal aspirates was measured. Infants received either zero (n = 5), one (n = 4), two (n = 15), or three (n = 3) doses of Survanta (100 mg/kg) when clinically indicated. MEASUREMENTS AND MAIN RESULTS: Using multiple regression analysis, the absolute synthesis rate (ASR) of surfactant PC from plasma glucose increased with 1.3 +/- 0.4 mg/kg/day per dose of Survanta (p = .007) (mean +/- SEM). The ASR of surfactant PC from glucose was increased by prenatal corticosteroid treatment with 1.3 +/- 0.4 mg/kg/day per dose corticosteroid (p = .004), and by the presence of a patent ductus arteriosus with 2.1 +/- 0.7 mg/ kg/day (p = .01). CONCLUSION: These data are reassuring and show for the first time in preterm infants that multiple doses of exogenous surfactant for RDS are tolerated well by the developing lung and stimulate endogenous surfactant synthesis. PMID- 11057791 TI - Outcome, functional autonomy, and quality of life of elderly patients with a long term intensive care unit stay. AB - OBJECTIVE: To examine the outcome, functional autonomy, and quality of life of elderly patients (> or = 70 yrs old) hospitalized for >30 days in an intensive care unit (ICU). DESIGN: Prospective cohort study. SETTING: A ten-bed, medical surgical ICU in a 460-bed, acute care, tertiary, university hospital. PATIENTS: A consecutive cohort of 75 patients, >70 yrs old, admitted to the ICU from January 1, 1993, to August 1, 1998, for >30 days. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Severity at admission and of the underlying disease was estimated according to the Simplified Acute Physiologic Score (SAPS II), the Organ Dysfunction and/or Infection (ODIN) score, the McCabe score, and the Knaus classification. Therapeutic intensity was measured through the French Omega scoring system. All patients were mechanically ventilated during their ICU stay. Outcome measurements were made by two cross-sectional studies using telephone interviews on the first week of September 1996 and 1998 with a questionnaire including measures of functional capacity by Katz's Activities of Daily Living, modified Patrick's Perceived Quality of Life score, and the Nottingham Health Profile. The survival rate was 67% in the ICU and 47% in the hospital. A total of 30 patients were alive and able to participate in at least one of the cross sectional studies. Independence in activities of daily living was decreased significantly after the ICU stay, except for feeding. However, most of the 30 patients remained independent (class A of the Activities of Daily Living index) with the possibility of going home. Perceived Quality of Life scores remained good, even if the patients estimated a decrease in their quality of life for health and memory. Return to society appeared promising regarding patient self respect and happiness with life. The estimated cost by survivor was of 55,272 EUR ($60,246 US). CONCLUSIONS: This study suggests that persistent high levels of ICU therapeutic intensity were associated with a reasonable hospital survival in elderly patients experiencing prolonged mechanical ventilatory support. These patients presented a moderate disability that influenced somewhat their perceived quality of life. These results are sufficient to justify prolonged ICU stays for elderly patients. PMID- 11057792 TI - Effects of maximizing oxygen delivery on morbidity and mortality in high-risk surgical patients. AB - OBJECTIVE: To evaluate the effects of maximizing the oxygen delivery on morbidity and mortality in patients >60 yrs of age and/or with chronic diseases of vital organs who underwent major elective surgery. DESIGN: Prospective, randomized, controlled trial. SETTING: A 24-bed general intensive care unit of a teaching hospital. PATIENTS: Thirty-seven high-risk patients who underwent major surgery. INTERVENTIONS: The hemodynamic and oxygen transport variables and outcomes in 18 patients (control group) treated to maintain normal values of oxygen delivery were compared with 19 patients (protocol group) treated to maintain "supranormal" values. Therapy in both groups consisted of volume expansion and, when necessary, dobutamine to reach target values, during the surgery and 24 hrs postoperatively. MEASUREMENTS AND MAIN RESULTS: We interrupted the study because of a significant difference in the 60-day mortality rate. The mortality rate in the control group was significantly higher when compared with the protocol group (9/18 [50%] vs. 3/19 [15.7%], p < .05). The prevalence of clinical and infectious complications was higher in the control group than in the protocol group (67% and 31% respectively; relative risk, 0.47; 95% confidence interval, 0.226-0.991; p < .05) and there was a trend toward more severe organ dysfunction in nonachievers patients (17/24 [71%] vs. 6/13 [46%], relative risk, 0.65; 95% confidence interval, 0.343-1.237; NS). CONCLUSION: Older patients with existing cardiorespiratory illness undergoing major surgery have a reduced morbidity and mortality when dobutamine is used to maximize oxygen transport. PMID- 11057793 TI - Changing pattern of organ dysfunction in early human sepsis is related to mortality. AB - OBJECTIVE: We examined the pattern of organ system dysfunction, the evolution of this pattern over time, and the relationship of these features to mortality in patients who had sepsis syndrome. DESIGN: Prospective, multicenter, observational study. SETTING: Intensive care units in tertiary referral teaching hospitals. PATIENTS: A total of 287 patients who had sepsis syndrome were prospectively identified in intensive care units. MATERIALS AND MEASUREMENTS: Cardiovascular, pulmonary, neurologic, coagulation, renal, and hepatic dysfunction were assessed at onset and on day 3 of sepsis syndrome. Organ dysfunction was classified as normal, mild, moderate, severe, and extreme dysfunction. We calculated the occurrence rate and associated 30-day mortality rate of organ dysfunction at the onset of sepsis syndrome. We then measured the change in organ dysfunction from onset to day 3 of sepsis syndrome and determined, for individual organ systems, the associated 30-day mortality rate. RESULTS: At the onset of sepsis syndrome, clinically significant pulmonary dysfunction was the most common organ failure, but was not related to 30-day mortality. Clinically significant cardiovascular, neurologic, coagulation, renal, and hepatic dysfunction were less common at the onset of sepsis syndrome but were significantly associated with the 30-day mortality rate. Worsening neurologic, coagulation, and renal dysfunction from onset to day 3 of sepsis syndrome were associated with significantly higher 30 day mortality than with improvement or no change in organ dysfunction. CONCLUSIONS: Increased mortality rate in sepsis syndrome is associated with a pattern characterized by failure of nonpulmonary organ systems and, in particular, worsening neurologic, coagulation, and renal dysfunction over the first 3 days. Although initial pulmonary dysfunction is common in patients with sepsis syndrome, it is not associated with an increased mortality rate. PMID- 11057794 TI - Pharmacokinetics and dosing regimen of meropenem in critically ill patients receiving continuous venovenous hemofiltration. AB - OBJECTIVE: To study the pharmacokinetics of meropenem in critically ill patients with acute renal failure receiving continuous venovenous hemofiltration (CWHF). DESIGN: Prospective, open-labeled study. SETTING: Medical intensive care unit of the University Medical Center Utrecht. PATIENTS: Five critically ill patients receiving CWHF for acute renal failure treated with meropenem for documented or suspected bacterial infection. INTERVENTION: All patients received meropenem (500 mg) administered intravenously every 12 hrs. Plasma samples and ultrafiltrate aliquots were collected during one dosing interval. MEASUREMENTS AND RESULTS: Mean age and body weight of the patients studied were 46.6 yrs (range, 28-61 yrs) and 85.8 kg (range, 70-100 kg), respectively. The following pharmacokinetic variables for meropenem were obtained: mean peak plasma concentration was 24.5 +/ 7.2 mg/L, mean trough plasma concentration was 3.0 +/- 0.9 mg/L, mean terminal elimination half-life was 6.37 +/- 1.96 hrs, mean total plasma clearance was 4.57 +/- 0.89 L/hr, mean CWHF clearance was 1.03 +/- 0.42 L/hr, mean nonrenal clearance was 3.54 +/- 1.06 L/hr, and mean volume of distribution was 0.37 +/- 0.15 L/kg. CONCLUSION: In critically ill patients with acute renal failure, nonrenal clearance became the main elimination route. CWHF substantially contributed to the clearance of meropenem (23% of mean total plasma clearance). We recommend meropenem to be dosed at 500 mg intravenously every 12 hrs in patients receiving CWHF, according to our operational characteristics. This dosing regimen resulted in adequate trough plasma levels for susceptible microorganisms. PMID- 11057795 TI - Effect of dopexamine on outcome after major abdominal surgery: a prospective, randomized, controlled multicenter study. European Multicenter Study Group on Dopexamine in Major Abdominal Surgery. AB - OBJECTIVE: To test the hypothesis that dopexamine reduces postoperative mortality and morbidity in high-risk, major abdominal surgery patients, when given to fluid resuscitated patients starting before the operation and continued for 24 hrs after surgery. DESIGN: Prospective, randomized, controlled, double-blind multicenter trial. SETTING: Intensive care units in 13 hospitals from six European countries. PATIENTS: A total of 412 patients with predefined high-risk criteria, undergoing major abdominal surgery with an expected duration of at least 1.5 hrs. INTERVENTIONS: The patients received placebo (n = 140), dopexamine at 0.5 microg/kg/min (n = 135), or dopexamine at 2.0 microg/kg/ min (n = 137) starting after preoperative hemodynamic stabilization and continued for 24 hrs after surgery. MEASUREMENTS AND MAIN RESULTS: The primary outcome variable was mortality at 28 days. Analysis was by intention to treat. Dopexamine had no effect on mortality (at 28 days, 13%, 7%, and 15%, for the groups receiving placebo, dopexamine at 0.5 microg/kg/ min, and dopexamine at 2.0 microg/kg/min, respectively), despite the expected dose-dependent hemodynamic responses. No effect was observed on the occurrence of organ dysfunction, duration of intensive care unit stay, or length of hospital stay. CONCLUSION: We conclude that dopexamine in doses that result in increased cardiac output and oxygen delivery after preoperative stabilization with fluids does not improve outcome after major abdominal surgery compared with fluids alone. Based on post hoc subgroup analysis and stratification according to the number of risk factors, we suggest that the concept should be further tested in patients at higher risk of complications or undergoing emergency surgery. PMID- 11057796 TI - Effect of mortality rate on the performance of the Acute Physiology and Chronic Health Evaluation II: a simulation study. AB - OBJECTIVE: To evaluate the impact of case mix variation on the performance of the Acute Physiology and Chronic Health Evaluation (APACHE) II using measures of calibration and discrimination. DESIGN: APACHE II data were collected prospectively at the surgical intensive care unit of the University of Vermont on all adult admissions over an 8-yr period (excluding cardiac surgical patients, burn patients, and patients < 16 yrs of age). The original case mix was systematically varied to create 2,000 different case mixes ranging in mortality between 5% and 18% using a computer-intensive resampling algorithm. The area under the receiver operating characteristic curve and the Hosmer-Lemeshow C statistic were derived for each of the simulated case mixes with bootstrapping. SETTING: The surgical intensive care unit at a 450-bed teaching hospital. PATIENTS: A group of 6,806 adult surgical patients excluding cardiac surgical patients and burn patients. MEASUREMENTS AND RESULTS: Simulated data sets were created from a database of patients treated at a single institution to test the hypothesis that the performance of APACHE II is stable across a clinically reasonable range of mortality rates. The discrimination and calibration of APACHE II varied with case mix. CONCLUSION: The discrimination of APACHE II is not independent of case mix. However, the variability of the Hosmer-Lemeshow statistic as a function of the case mix may simply reflect the limitations of this goodness of fit statistic to assess model calibration. Because the discrimination of APACHE II is a function of case mix, caution should be exercised when using APACHE II-based adjusted mortality rates to compare intensive care units with widely divergent case mixes. PMID- 11057797 TI - Inhaled nitric oxide does not enhance lipid peroxidation in patients with acute respiratory distress syndrome. AB - OBJECTIVE: To investigate whether inhaled nitric oxide (NO) enhances pulmonary lipid peroxidation as indicated by arterial blood levels of malondialdehyde, hexanal, and pentanal in patients with acute respiratory distress syndrome (ARDS). DESIGN: Prospective, nonrandomized, controlled trial. SETTING: Surgical intensive care unit in a university hospital. PATIENTS: Twenty-five patients with ARDS, as defined by the American-European Consensus Conference, and a PaO2/FIO2 < or = 170 mm Hg were enrolled in the study. Four healthy subjects were studied as controls. INTERVENTIONS: On enrollment of the patients in the study, a dose response test with increasing concentrations of inhaled NO (0, 2, 10, 40, 0 ppm) was performed. Patients who showed an increase of >20% in PaO2 were designated as responders and all others as nonresponders. In responders, this dose-response test was followed by 24 hrs of continuous treatment with inhaled NO at the best NO concentration determined during the dose-response test, whereas nonresponders received standard care. For healthy volunteers, the dose-response test took the form of spontaneous breathing of the same NO concentrations. MEASUREMENTS AND MAIN RESULTS: Eighteen patients (72%) showed an increase of >20% in PaO2 during the dose-response test. This significant improvement in arterial oxygenation in responders led to a significant reduction in FIO2 (responders, 0.73 +/- 0.05 vs. nonresponders, 0.89 +/- 0.05) after 24 hrs of therapy. On enrollment, arterial blood concentrations of malondialdehyde, hexanal, and pentanal were significantly higher than those of healthy volunteers. In addition, arterial concentrations of hexanal and pentanal exceeded mixed venous levels two- to ten-fold. Inhalation of NO did not significantly alter these blood concentrations either during the dose response test or during 24 hrs of therapy. CONCLUSIONS: In patients with ARDS, malondialdehyde, hexanal, and pentanal were significantly elevated, indicating lipid peroxidation. Lipid peroxidation was not further affected by inhalation of NO. PMID- 11057798 TI - Cervical spine clearance and neck extension during percutaneous tracheostomy in trauma patients. AB - INTRODUCTION: The lack of cervical spine clearance and inability to extend the neck are assumed to be relative contraindications for percutaneous tracheostomy. OBJECTIVE: To determine the necessity of cervical spine clearance and neck extension in trauma patients receiving percutaneous tracheostomy. DESIGN: Prospective analysis of case series from August 1, 1995 to August 31, 1998. SETTING: A university-based Level I trauma center. PATIENTS: A total of 88 consecutive trauma patients receiving percutaneous tracheostomy. Patients were divided into two groups based on the radiographic or clinical status of their cervical spine: cleared and noncleared. RESULTS: The overall success and complication rate were 99% (87/88) and 11% (10/88), respectively. There were no procedure-related deaths. The cleared group consisted of 60 patients; three patients in this group who had "bull" or "thick" necks did not have full neck extension during percutaneous tracheostomy. The noncleared group consisted of 28 patients, 13 of which had known cervical spine fractures; 27 noncleared patients were maintained in the neutral position (no extension) during percutaneous tracheostomy, whereas one patient with low suspicion of spinal injury was partially extended. Of the 13 patients with cervical spine fractures, six patients had been stabilized with a halo or operative fixation, and seven patients were stabilized with a cervical collar at the time of percutaneous tracheostomy. The success rate was 100% (60/60) for the cleared group compared with 96% (27/28) for the noncleared group (p > .05). The complication rate was 13% (8/60) for the cleared group compared with 7.1% (2/28) for the noncleared group (p > .05). We had a 100% success rate and no complications in the seven patients with cervical spine injury who were stabilized with a cervical collar. No patient had spinal cord injury caused by percutaneous tracheostomy. CONCLUSION: Percutaneous tracheostomy can be safely performed in trauma patients without cervical spine clearance and neck extension, including patients with stabilized cervical spine or spinal cord injury. PMID- 11057799 TI - Biochemical changes after trauma and skeletal surgery of the lower extremity: quantification of the operative burden. AB - OBJECTIVE: To quantify changes in variables of inflammation, coagulation, and fibrinolysis in blunt trauma patients with lower extremity fractures who underwent different types of surgical procedures. DESIGN: Prospective, cohort study. SETTING: Level I university trauma center. PATIENTS: We allocated 83 blunt trauma patients in stable condition and 22 patients eligible for elective hip replacement to four treatment groups. INTERVENTIONS: In 34 multiply traumatized patients with femoral fracture (group PTFF) and in 28 patients with an isolated femoral fracture (group IFF), primary unreamed intramedullary nailing for stabilization of the femoral shaft fracture was performed. In 22 patients, an elective uncemented total hip arthroplasty (group THA) was inserted for osteoarthritis, and in 21 control patients, an isolated ankle fracture (group AF) was acutely stabilized. MEASUREMENTS AND MAIN RESULTS: From serially sampled central venous blood, the perioperative concentrations of interleukin (IL)-6, of tumor necrosis factor-alpha, of prothrombin fragments 1 + 2, and of D-dimer cross linked fibrin degradation products were evaluated. Intramedullary instrumentation for an isolated femur fracture caused a significant perioperative increase in the concentrations of IL-6 (preoperative IL-6, 52 +/- 12 pg/mL; IL-6 30 mins postinsertion, 78 +/- 14 pg/mL; p = .02). This increase was comparable with group THA (preoperative IL-6, 46 +/- 16 pg/mL; IL-6 30 mins postinsertion, 67 +/- 11 pg/mL; p = .03). A positive correlation occurred between both groups (r = .83, p < .0004). Multiple trauma patients demonstrated significantly (p = .0002) higher IL-6 concentrations than all other groups throughout the study period and showed a significant increase after femoral nailing (preoperative IL-6, 570 +/- 21 pg/mL; IL-6 30 mins postinsertion, 690 +/- 24 pg/mL; p = .003), whereas no perioperative change was seen in group AF. The highest IL-6 increases were associated with a longer ventilation time (group PTFF) and a longer period of positive fluid balances (groups PTFF, IFF, THA). The coagulatory variables demonstrated similar perioperative increases in groups IFF and THA, but not in groups PTFF and AF. The IL-6 concentrations and the prothrombin fragments 1 + 2 concentrations correlated between groups THA and IFF at 30 mins and at 1 hr after surgery (r2 = .64, p < .02). In all patients the clinical variables were stable perioperatively. CONCLUSIONS: Major surgery of the lower extremity causes changes to the inflammatory, fibrinolytic, and coagulatory cascades in patients with stable cardiopulmonary function. The inflammatory response induced by femoral nailing is biochemically comparable to that induced by uncemented total hip arthroplasty. In multiple trauma patients, increases, which occurred in addition to those induced by the initial trauma, were measured. Definitive primary femoral stabilization by intramedullary nailing imposes an additional burden to the patient with blunt trauma. A careful preoperative investigation is required to evaluate whether primary definitive stabilization can be performed safely. PMID- 11057800 TI - Gastric tonometry in patients with cardiogenic shock and intra-aortic balloon counterpulsation. AB - OBJECTIVE: To study the course of gastric regional PCO2 (PrCO2) in patients with cardiogenic shock requiring intra-aortic balloon (IAB) counterpulsation and the prognostic value of PrCO2 in this patient population. DESIGN: A prospective, observational clinical study. SETTING: Medical intensive care unit in a university hospital. PATIENTS: Twenty-six consecutive patients with cardiogenic shock requiring mechanical support with an IAB counterpulsation undergoing mechanical ventilation INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULT: Hemodynamic variables, tonometric variables, arterial blood gases, and arterial lactate were assessed before insertion of IAB (baseline), and 1, 2, 3, 8, 16, 24, and 48 hrs thereafter. A subset of these patients (n = 14) were studied just before and 1, 8, 24, and 32 hrs after IAB removal; 12/26 patients (46.2%) died. Cardiac index increased from baseline to 1 hr after insertion of IAB (1.7 +/- 0.3 to 2.6 +/- 0.8 L/min/m2, p < .05). PrCO2 did not change between admission (47 +/- 13 torr [6.3 +/- 1.7 kPa]) and 8 hrs after placement of IAB but increased to 63 +/- 22 torr (8.4 +/- 2.9 kPa) at 16 hrs (p < .05) without any further alteration until 48 hrs. CO2 gap showed a similar pattern with 15 +/- 11 torr (2.0 +/- 1.5 kPa) at baseline and an increase to 28 +/- 22 torr (3.7 +/- 2.9 kPa) 16 hrs later. PrCO2 and CO2 gap remained at high levels (59 +/- 11 torr [7.7 +/- 1.5 kPa] and 22 +/- 10 torr [2.9 +/- 1.3 kPa], respectively), before IAB removal without further improvement or deterioration thereafter. PrCO2 values showed no difference between survivors and nonsurvivors at any time point. CONCLUSION: Patients with cardiogenic shock developed high PrCO2 within the first 24 hrs, which reflects gastric mucosal ischemia. Persistently high levels of PrCO2 were indicative for prolonged hypoperfusion of the gut. Gastric tonometry failed to discriminate between survivors and nonsurvivors. PMID- 11057801 TI - Inadequate treatment of nosocomial infections is associated with certain empiric antibiotic choices. AB - OBJECTIVE: The purpose of this study was to determine the impact of scheduled changes of antibiotic classes, used for the empirical treatment of suspected or documented Gram-negative bacterial infections, on the occurrence of inadequate antimicrobial treatment of nosocomial infections. DESIGN: Prospective observational study. SETTING: Medical (19-bed) and surgical (18-bed) intensive care units in an urban teaching hospital. PATIENTS: A total of 3,668 patients requiring intensive care unit admission were prospectively evaluated during three consecutive time periods. INTERVENTIONS: During each time period, one antibiotic class was selected for the empirical treatment of Gram-negative bacterial infections as follows: time period 1 (baseline period) (1,323 patients), ceftazidime; time period 2 (1,243 patients), ciprofloxacin; and time period 3 (1,102 patients), cefepime. MEASUREMENTS AND MAIN RESULTS: The overall administration of inadequate antimicrobial treatment for nosocomial infections decreased during the course of the study (6.1%, 4.7%, and 4.5%; p = .15). This was primarily because of a statistically significant decrease in the administration of inadequate antibiotic treatment for Gram-negative bacterial infections (4.4%, 2.1%, and 1.6%; p < .001). There were no statistically significant differences in the overall hospital mortality rate among the three time periods (15.6%, 16.4%, and 16.2%; p = .828) despite a significant increase in severity of illness as measured with Acute Physiology and Chronic Health Evaluation (APACHE) II scores (15.3 +/- 7.6, 15.7 +/- 8.0, and 20.7 +/- 8.6; p < .001). The hospital mortality rate decreased significantly during time period 3 (20.6%) compared with time period 1 (28.4%; p < .001) and time period 2 (29.5%; p < .001) for patients with an APACHE II score > or = 15. CONCLUSIONS: These data suggest that scheduled changes of antibiotic classes for the empirical treatment of Gram-negative bacterial infections can reduce the occurrence of inadequate antibiotic treatment for nosocomial infections. Reducing inadequate antibiotic administration may improve the outcomes of critically ill patients with APACHE II scores > or = 15. PMID- 11057802 TI - Intensive care unit length of stay: recent changes and future challenges. AB - OBJECTIVE: To compare case-mix adjusted intensive care unit (ICU) length of stay for critically ill patients with a variety of medical and surgical diagnoses during a 5-yr interval. DESIGN: Nonrandomized cohort study. SETTING: A total of 42 ICUs at 40 US hospitals during 1988-1990 and 285 ICUs at 161 US hospitals during 1993-1996. PATIENTS: A total of 17,105 consecutive ICU admissions during 1988-1990 and 38,888 consecutive ICU admissions during 1993-1996. MEASUREMENTS AND MAIN RESULTS: We used patient demographic and clinical characteristics to compare observed and predicted ICU length of stay and hospital mortality. Outcomes for patients studied during 1993-1996 were predicted using multivariable models that were developed and cross-validated using the 1988-1990 database. The mean observed hospital length of stay decreased by 3 days (from 14.8 days during 1988-1990 to 11.8 days during 1993-1996), but the mean observed ICU length of stay remained similar (4.70 vs. 4.53 days). After adjusting for patient and institutional differences, the mean predicted 1993-1996 ICU stay was 4.64 days. Thus, the mean-adjusted ICU stay decreased by 0.11 days during this 5-yr interval (T-statistic, 4.35; p < .001). The adjusted mean ICU length of stay was not changed for patients with 49 (75%) of the 65 ICU admission diagnoses. In contrast, the mean observed hospital length of stay was significantly shorter for 47 (72%) of the 65 admission diagnoses, and no ICU admission diagnosis was associated with a longer hospital stay. Aggregate risk-adjusted hospital mortality during 1993-1996 (12.35%) was not significantly different during 1988 1990 (12.27%, p = .54). CONCLUSIONS: For patients admitted to ICUs, the pressures associated with a decrease in hospital length of stay do not seem to have influenced the duration of ICU stay. Because of the high cost of intensive care, reduction in ICU stay may become a target for future cost-cutting efforts. PMID- 11057803 TI - Auto-positive end-expiratory pressure during tracheal gas insufflation: testing a hypothetical model. AB - OBJECTIVE: The major benefit of tracheal gas insufflation (TGI) is an increase in CO2 elimination efficiency by removal of CO2 from the anatomical deadspace. In conjunction with mechanical ventilation, TGI may also alter variables that affect CO2 elimination, such as minute ventilation and peak airway pressure (peak Paw) and cause the development of auto-positive end-expiratory pressure (auto-PEEP). We tested the hypothesis that TGI-induced auto-PEEP alters ventilatory variables. We predicted that TGI-induced auto-PEEP offsets the beneficial effects of TGI on CO2 elimination and that keeping total PEEP (ventilator PEEP + auto-PEEP) constant enhances the CO2 elimination efficiency afforded by TGI. DESIGN: Prospective study of two series of patients with acute respiratory distress syndrome receiving mechanical ventilation. SETTING: Intensive care units at a university medical center. PATIENTS: Each series consisted of eight sequential hypercapnic patients. INTERVENTIONS: In series 1, we examined the effect of continuous TGI at 0 and 10 L/min on PaCO2, without compensating for the development of auto-PEEP. In series 2, we examined this same effect of continuous TGI while reducing ventilator PEEP to keep total PEEP constant. TGI-induced auto PEEP was calculated based on dynamic compliance measurements during zero TGI flow conditions (deltaV/deltaP) after averaging the two baseline values for peak Paw and tidal volume and assuming compliance did not change between the zero TGI and TGI flow conditions (deltaVTGI/deltaPTGI). MEASUREMENTS AND MAIN RESULTS: In series 1, total PEEP increased from 13.2 +/- 3.2 cm H2O to 17.8 +/- 3.5 cm H2O without compensation for auto-PEEP (p = .01). PaCO2 decreased (p = .03) from 56.2 +/- 10.6 mm Hg (zero TGI) to 52.9 +/- 9.3 mm Hg (TGI at 10 L/min), a 6% decrement. In series 2, total PEEP was unchanged (p = NS). PaCO2 decreased (p = .03) from 59.5 +/- 10.4 mm Hg (zero TGI) to 52.2 +/- 8.3 mm Hg (TGI at 10 L/min), a 12% decrement. There was no significant change in PaO2; there were no untoward hemodynamic effects in either series. CONCLUSIONS: These data are consistent with the hypothesis that mechanical ventilation + TGI causes an increase in auto-PEEP that can blunt CO2 elimination. In addition to the ventilator modifications necessary to keep ventilatory variables constant when TGI is used, it is also necessary to reduce ventilator PEEP to keep total PEEP constant and further enhance CO2 elimination efficiency. PMID- 11057804 TI - Analysis of static pulmonary mechanics helps to identify functional defects in survivors of acute respiratory distress syndrome. AB - OBJECTIVE: To assess the utility of static lung pressure-volume measurements in identifying and categorizing pulmonary function test defects in survivors of acute respiratory distress syndrome (ARDS). DESIGN: Cross-sectional. SETTING: Pulmonary function laboratory at a tertiary referral hospital in northern India. PATIENTS: Six survivors of ARDS reporting for their first follow-up visit after discharge. MEASUREMENTS AND MAIN RESULTS: Spirometry and whole body plethysmography were performed to evaluate lung volumes and to collect static lung pressure-volume data; the latter were subjected to monoexponential analysis. Three patients had a restrictive ventilatory defect as evidenced by diminished vital capacity and total lung capacity, but only one had reduced static lung compliance. The other two patients had reduced recoil pressure at total lung capacity, suggestive of respiratory muscle weakness. Two other patients with normal lung volumes had reduced static lung compliance. Exponential analysis of pressure-volume data in the three patients with reduced static lung compliance was consistent with reduced distensibility and loss of lung units in one patient each. Additionally, two patients had high values for shape constant of the exponential curve, indicative of air space distention. CONCLUSIONS: Detailed analysis of static pressure-volume data can identify pulmonary function abnormality and categorize the dominant mechanism responsible for restrictive ventilatory defects in survivors of ARDS, even in patients with normal lung volumes. In addition to lung fibrosis, neuromuscular weakness also contributes to decline in pulmonary function. PMID- 11057805 TI - Effects of dobutamine on splanchnic tissue perfusion during partial superior mesenteric artery occlusion. AB - OBJECTIVE: To evaluate the effects of dobutamine and fluid treatment on splanchnic hemodynamics and tissue oxygenation during partial superior mesenteric artery occlusion. DESIGN: Prospective, open randomized, full-factorial design. SETTING: University research laboratory. SUBJECTS: Forty-eight female pigs. INTERVENTIONS: In 24 anesthetized pigs (ischemic group), superior mesenteric artery (SMA) blood flow was reduced to 30% from the baseline for 120 mins; 24 pigs (sham group) served as nonischemic controls. The animals were further assigned into four treatment arms. In the control arm, the animals were administered only basic fluid therapy. In the fluid therapy arm, pulmonary artery occlusion pressure was maintained at 10 mm Hg with fluids. In the dobutamine treatment arm, dobutamine hydrochloride was infused at a dose of 10 microg/min/kg. In the combined dobutamine-fluid therapy arm, dobutamine at 10 microg/min/kg was administered and pulmonary artery occlusion pressure was maintained at 10 mm Hg with fluids. MEASUREMENTS AND MAIN RESULTS: Systemic and regional hemodynamics and oxygen transport, as well as jejunal intramucosal pH, intramucosal-arterial PCO2 gradient, and portal venous-arterial lactate gradient were measured. Ischemia did not modify the effects of fluids or dobutamine on systemic hemodynamics and oxygen transport. Dobutamine-treated animals had a higher cardiac index compared with control animals (218 +/- 22 vs. 135 +/- 13 mL/min/kg; p = .012), and the effect was enhanced when dobutamine was combined with fluid treatment (365 +/- 23 mL/ min/kg; p = .019). Fluid treatment alone did not influence cardiac index, whereas it increased SMA blood flow compared with control groups (15 +/- 2 vs. 12 +/- 2 mL/min/kg; p = .023). Dobutamine also decreased the proportion of SMA blood flow of cardiac output compared with control groups (6 +/- 1 vs. 9% +/- 1%; p = .024). Other treatments had no effect on SMA blood flow. Ischemia increased intramucosal-arterial Pco2 gradient to 54.8 +/- 10.7 torr (7.31 +/- 1.43 kPa) (p = .002 vs. sham control) and decreased intramucosal pH to 7.13 +/- 0.06 (p = .028 vs. sham control). In the ischemic animals, dobutamine without fluid therapy reduced intramucosal pH further to 7.00 +/- 0.09 (p = .023 vs. ischemic control) and increased portal venous-arterial lactate gradient (p = .033). CONCLUSIONS: Dobutamine alone worsened splanchnic tissue perfusion during partial superior mesenteric artery occlusion. As compared with fluid treatment alone, the combination of fluid and dobutamine therapy did not improve tissue perfusion. PMID- 11057806 TI - Escherichia coli endotoxin reduces cytochrome aa3 redox status in pig skeletal muscle. AB - OBJECTIVE: To assess the effect of endotoxin on cytochrome aa3 (Caa3) redox status in a controlled blood flow preparation of pig isolated hindlimb, at a constant oxygen delivery (Do2limb) (constant flow period) and during progressive ischemia (decreasing flow period). DESIGN: Randomized, controlled experimental study. SETTING: University hospital experimental laboratory. SUBJECTS: Ten piglets. INTERVENTIONS: Hindlimb blood flow was restricted to the femoral vessels. The arterial femoral blood flow coming from the carotid artery was controlled by a roller occlusive pump. The femoral venous blood flow was returned to the jugular vein. During the first 100 mins, the hindlimb blood flow was maintained at a normal level and then decreased stepwise. Animals were randomized to receive 150 microg/kg endotoxin lipopolysaccharide (LPS; n = 5) or saline (control; n = 5). MEASUREMENTS AND MAIN RESULTS: Hindlimb muscle Caa3 redox status was monitored by near-infrared spectroscopy. Hindlimb Do2limb and oxygen consumption (Vo2limb) were calculated. In the LPS group, a rapid reduction of Caa3 redox status was observed after LPS administration, whereas the hindlimb blood flow remained normal with no change in Do2limb and Vo2limb. A progressive simultaneous decrease in Do2limb and Vo2limb was observed during the decreasing flow period with no further reduction in Caa3 redox status. In the control group, no change was observed in Caa3, Do2limb, or Vo2limb during the constant flow period. During the decreasing flow period, Caa3 redox status was reduced as Do2limb and Vo2limb decreased. CONCLUSION: Our results suggest that endotoxin may induce a reduction of Caa3 redox status independently of Do2 and Vo2. PMID- 11057807 TI - Does early infusion of red blood cells after trauma and hemorrhage improve organ functions? AB - OBJECTIVE: Early management of trauma victims includes control of bleeding and rapid restoration of intravascular volume. However, it remains controversial whether infusion of blood products is superior to crystalloids alone. Therefore, it was the aim of the present study to determine whether resuscitation with red blood cells plus lactated Ringer's solution (RL) is more effective than RL alone in improving the cardiovascular and hepatocellular functions after trauma and severe hemorrhage. DESIGN: Prospective study. SETTING: Laboratory. SUBJECTS: Sprague-Dawley rats. INTERVENTIONS AND MEASUREMENTS: Male adult rats were anesthetized and underwent a laparotomy to induce tissue trauma before hemorrhage. The animals were then bled to and maintained at a mean arterial pressure of 40 mm Hg until 40% of the maximal bleed-out (MB) volume was returned in the form of RL, and were then resuscitated with either four times the volume of MB with RL or washed red blood cells (RBC) (-45% the volume of MB) in three times the volume of RL over 60 mins. Various in vivo heart performance variables, cardiac output, and hepatocellular function (ie, the maximum velocity and the overall efficiency of indocyanine green clearance) were determined at 4 hrs after resuscitation. Hemoglobin, systemic oxygen delivery, circulating blood volume, and plasma levels of interleukin-6 were also measured. MAIN RESULTS: At 4 hrs after RL resuscitation, heart performance, cardiac output and hepatocellular function were significantly depressed and plasma levels of interleukin-6 were significantly increased. Although infusion of RBC significantly increased mean arterial pressure, hemoglobin, and oxygen delivery compared with animals resuscitated with RL only, infusion of RBC did not further improve the depressed cardiovascular and hepatocellular functions under such conditions. CONCLUSION: Because infusion of RBC and RL resuscitation do not improve organ functions compared with RL resuscitation without RBC, it appears that pharmacologic agents in addition to fluid resuscitation are needed to restore cardiovascular and hepatocellular functions after trauma and hemorrhage. PMID- 11057808 TI - Cerebral blood flow and metabolism during and after prolonged hypercapnia in newborn lambs. AB - OBJECTIVE: To study the effects of prolonged (6 hrs) hypercapnia on cerebral blood flow and cerebral metabolism in newborn lambs and to evaluate the effects on cerebral blood flow and cerebral metabolism on return to normocapnia after prolonged hypercapnia. DESIGN: Animal studies, using the newborn lamb, with comparison to control group. SUBJECTS: Newborn lambs of mixed breed, 1-7 days of age, were used for the study. Two groups of animals were studied: a hypercapnic group (n = 10) and a normocapnic control group (n = 5). SETTING: Work was conducted in the research laboratories at Children's National Medical Center, Washington, DC. INTERVENTIONS: Animals were anesthetized with pentobarbital, intubated, paralyzed, and mechanically ventilated. After baseline measurements were made, CO2 was blended into the ventilator gas until a PaCO2 of 75-80 torr (10-10.6 kPa) was obtained. Measurements were made 1 hr after the desired PaCO2 was achieved and after 6 hrs of hypercapnia. After 6 hrs of hypercapnia, the ventilator gas was returned to the baseline value, that is, normocapnia. Measurements were made 30, 60, and 90 mins after PaCO2 returned to baseline. MEASUREMENTS: Six measurements were made during the study. For each measurement, blood samples were drawn from the sagittal sinus and brachiocephalic artery catheters and were analyzed for pH, hemoglobin concentration, oxygen saturation, and blood gas values. Cerebral blood flow (CBF) was measured by using the radiolabeled microsphere technique. Cerebral oxygen consumption, fractional oxygen extraction, and oxygen transport values were calculated at each study period. MAIN RESULTS: Increasing PaCO2 from 37 +/- 3 torr to 78 +/- 6 torr (4.9 +/- 0.4 kPa to 10.3 +/- 0.8 kPa) for 1 hr increased CBF by 355%. After 6 hrs of PaCO2 at 78 +/- 3 torr (10.3 +/- 0.4 kPa), CBF remained 195% above baseline. At 30 mins of normocapnia, CBF had returned to baseline and remained at baseline until the conclusion of the study, a total of 90 mins of normocapnia. Cerebral oxygen consumption did not change during hypercapnia or with return to normocapnia. Oxygen transport increased 331% above baseline after 1 hr of hypercapnia and stayed 180% above baseline after 6 hrs of hypercapnia. Fractional oxygen extraction decreased by 55% at 1 hr of hypercapnia and stayed 39% below baseline at 6 hrs of hypercapnia. CONCLUSIONS: Healthy lambs seem to tolerate undergoing hypercapnia for 6 hrs with a return to normocapnia. The return to baseline of CBF and cerebral metabolism at normocapnia seen in our study with lambs may explain why prolonged hypercapnia appears to be well tolerated in mechanically ventilated patients. If these results can be extrapolated to human subjects, our study in lambs supports evidence that patients who have undergone permissive hypercapnia seem to be neurologically unaffected. PMID- 11057809 TI - Delayed, spontaneous hypothermia reduces neuronal damage after asphyxial cardiac arrest in rats. AB - OBJECTIVE: Core temperature is reduced spontaneously after asphyxial cardiac arrest in rats. To determine whether spontaneous hypothermia influences neurologic damage after asphyxial arrest, we compared neurologic outcome in rats permitted to develop spontaneous hypothermia vs. rats managed with controlled normothermia. INTERVENTIONS: Male Sprague-Dawley rats were asphyxiated for 8 mins and resuscitated. After extubation, a cohort of rats was managed with controlled normothermia (CN) by placement in a servo-controlled incubator set to maintain rectal temperature at 37.4 degrees C for 48 hrs. CN rats were compared with permissive hypothermia (PH) rats that were returned to an ambient temperature environment after extubation. Rats were killed at either 72 hrs (PH72hr, n = 14; CN72hr, n = 9) or 6 wks (PH6wk, n = 6, CN6wk, n = 6) after resuscitation. PH72 rats were historic controls for the CN72 rats, whereas PH6 and CN6 rats were randomized and studied contemporaneously. MEASUREMENTS: A clinical neurodeficit score (NDS) was determined daily. A pathologist blinded to group scored 40 hematoxylin and eosin -stained brain regions for damage by using a 5-point scale (0 = none, 5 = severe). Quantitative analysis of CA1 hippocampus injury was performed by counting normal-appearing neurons in a defined subsection of CA1. MAIN RESULTS: Mean rectal temperatures measured in the PH6wk rats (n = 6) were 36.9, 34.8, 35.5, 36.7, and 37.4 degrees C at 2, 8, 12, 24, and 36 hrs, respectively. Mortality rate (before termination) was lower in PH compared with CN (0/20 vs. 7/15; p < .005). PH demonstrated a more favorable progression of NDS (p = .04) and less weight loss (p < .005) compared with CN. Median histopathology scores were lower (less damage) in PH72hr vs. CN72hr for temporal cortex (0 vs. 2.5), parietal cortex (0 vs. 2), thalamus (0 vs. 3), CA1 hippocampus (1.5 vs. 4.5), CA2 hippocampus (0 vs. 3.5), subiculum (0 vs. 4), and cerebellar Purkinje cell layer (2 vs. 4) (all p < .05). There was almost complete loss of normal appearing CA1 neurons in CN72hr rats (6 +/- 2 [mean +/- SD] normal neurons compared with 109 +/- 12 in naive controls). In contrast, PH72hr rats demonstrated marked protection (97 +/- 23 normal-appearing neurons) that was still evident, although attenuated, at 6 wks (42 +/- 24 normal-appearing neurons, PH6wk). CONCLUSION: Rats resuscitated from asphyxial cardiac arrest develop delayed, mild to moderate, prolonged hypothermia that is neuroprotective. PMID- 11057810 TI - Vasopressin-mediated adrenocorticotropin release increases plasma cortisol concentrations during cardiopulmonary resuscitation. AB - OBJECTIVE: Vasopressin is a possible stimulus for both adrenocorticotropin (ACTH) and endothelin-1 release. The aim of this study was to compare plasma concentrations of ACTH, cortisol, and endothelin-1 after epinephrine or vasopressin administration in an experimental animal model of cardiopulmonary resuscitation (CPR). DESIGN: Prospective, randomized, controlled animal study. SETTING: A university research laboratory. SUBJECTS: Fourteen 12- to 14-wk-old domestic pigs. INTERVENTIONS: After 4 mins of cardiac arrest and 3 mins of external chest compression, the pigs were randomly assigned to receive either 0.045 mg/kg epinephrine (n = 7) or 0.4 units/kg vasopressin (n = 7). At 5 mins after drug administration, defibrillation was attempted. MEASUREMENTS AND MAIN RESULTS: Coronary perfusion pressure, ACTH, cortisol, and endothelin-1 were measured before cardiocirculatory arrest, during CPR before drug administration, and at 90 secs and 5 mins after drug administration. Coronary perfusion pressure was comparable between groups. All seven animals in the vasopressin group survived, but only one pig in the epinephrine group survived (p = .005). ACTH and cortisol concentrations remained unchanged in epinephrine-treated animals, but increased significantly after vasopressin administration and were significantly higher than in epinephrine-treated animals 5 mins after drug administration. Endothelin-1 concentrations remained unchanged during the study period and were comparable between both groups. CONCLUSIONS: Vasopressin is a potent stimulus for ACTH secretion, but does not trigger endothelin-1 release from vascular cells during cardiac arrest and CPR. The increased plasma cortisol concentrations caused by the enhanced ACTH release after vasopressin may be one factor contributing to the improved outcome repeatedly observed with vasopressin in animal models of CPR. PMID- 11057811 TI - Electroencephalogram silence ratio for early outcome prognosis in severe head trauma. AB - OBJECTIVE: To introduce the electroencephalogram silence-ratio (ESR) as a variable derived from mathematically processed electroencephalogram for early outcome prognosis in patients with severe head trauma and to comparatively assess sensitivity, specificity and predictive value vs. somatosensory evoked potentials and brainstem auditory evoked potentials. DESIGN: Prospective, interventional study. SETTING: Intensive care unit of a university hospital. PATIENTS: A total of 32 adults with severe acute head trauma (Glasgow Coma Scale score < or = 8). METHODS AND MAIN RESULTS: In all patients, electroencephalographic recording was continuously performed by frontomastoid electrode montage for 24-96 hrs after admission to the ICU. The data were subsequently computed by fast Fourier analysis and the ESR (intervals of suppression as periods >240 msecs during which the electroencephalographic voltage did not exceed 5 microV) was displayed and recorded on a computer for further evaluation. Somatosensory evoked potentials and brainstem auditory evoked potentials were elicited during the first 2 days after admission. Outcome evaluation was performed 6 months after trauma using the Glasgow Outcome Scale and the Rappaport Disability Rating Scale. After careful artifact exclusion, the ESR depicted the highest sensitivity, specificity, and positive predictive value compared with evoked potentials. Even a highly significant correlation between outcome and ESR was found (p < .0001). CONCLUSION: The ESR is a valuable variable showing a high reliability with respect to outcome prediction in severe head trauma with a higher predictive value than short latency somatosensory evoked potentials. Evidence exists that the ESR provides at least partial information regarding adequate cerebral oxygen delivery. PMID- 11057812 TI - Nosocomial pneumonia in the pediatric trauma patient: a single center's experience. AB - OBJECTIVES: To evaluate a single center's experience with the frequency rate, patterns of occurrence, and impact on outcome of nosocomial pneumonia in the critically injured child. DESIGN: Retrospective review of prospectively collected data. SETTING: Level I university trauma center with a pediatric trauma intensive care unit. PATIENTS: A total of 523 consecutive critically injured children admitted to the pediatric intensive care unit during an 80-month interval. MEASUREMENTS AND RESULTS: Thirty-five episodes of nosocomial pneumonia were identified in 29 children (frequency rate of 5.5%). The mean age of the children was 9.2 yrs, and the mean Injury Severity Score was 27 +/- 9. In 91% of patients (26 children), nosocomial pneumonia was associated with mechanical ventilation. This represented a 13% frequency rate in injured children who were ventilated during the study period. The most common organisms recovered were Staphylococcus aureus (21%), Haemophilus influenzae (19%), Pseudomonas (11%), and Enterobacter (11%). Early pneumonia (diagnosed < or = 7 days after injury) was predominantly caused by Haemophilus species. In contrast, Enterobacter and/or Pseudomonas were isolated primarily in late pneumonia (diagnosed >7 days after injury). Staphylococcus was prominent throughout the hospitalization. Overall, children with nosocomial pneumonia were more severely injured (Injury Severity Score 27 vs. 17, p < .001) and had a longer hospital stay (26 vs. 7 days, p < .001). Despite this, mortality (6.9% vs. 7.9%, p = NS) was not significantly different from injured children without pneumonia. CONCLUSIONS: In this study of a single pediatric trauma center, nosocomial pneumonia occurred in a small but significant percentage of injured children. The frequency rate increased two- to three-fold with mechanical ventilation. Microbiology varied with day of onset. In contrast to the adult, mortality did not seem to be significantly altered by this complication. Analysis of additional pediatric trauma centers is encouraged to confirm these characteristics of nosocomial pneumonia in the injured child. PMID- 11057813 TI - Evaluating the frequency rate of hypomagnesemia in critically ill pediatric patients by using multiple regression analysis and a computer-based neural network. AB - OBJECTIVES: To determine the frequency rate of hypomagnesemia in patients admitted to the pediatric intensive care unit (ICU), and to identify subsets of patients (grouped by disease) who are at greatest risk of hypomagnesemia. We also compared a neural network model with multiple regression analysis to identify independent variables that would correlate with hypomagnesemia and to predict serum magnesium values in critically ill pediatric patients overall. DESIGN: Prospective, multicenter study. SETTING: Tertiary level medical/surgical pediatric ICUs. PATIENTS: Data were obtained at admission to the pediatric ICU for 463 patients from newborn to 18 yrs old who were admitted with a variety of surgical and nonsurgical conditions. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Total serum magnesium values were obtained within the first 24 hrs after admission in 463 pediatric patients admitted to four pediatric ICUs. Hypomagnesemia (defined as total serum magnesium <0.75 mmol/L) was found in 51 (11%) of the 463 patients, with the highest frequency rate (72%) and lowest mean serum magnesium level (0.66 +/- 0.17 mmol/L) in patients admitted after surgery with extensive osseous involvement (spinal fusion and craniofacial reconstruction). To determine whether hypomagnesemia could be predicted on the basis of other laboratory and clinical criteria, multiple regression analysis was performed and showed age, weight, and albumin levels weakly associated (r2 = .14, p < .001) with magnesium levels within the different diagnostic groups. These data were used to produce a mathematical model able to predict magnesium levels within 5% of the actual values in 23% of patients. A neural network was also created to compare its predictive capabilities to those of the multiple regression model. Once trained on a random subset (85%) of the patient population, the neural network was able to predict magnesium levels to within 5% of actual values for 88% of the remaining 15% of patients, comparing favorably with the predictions derived from the multiple regression model. CONCLUSIONS: Hypomagnesemia is not uncommon (11%) in critically ill pediatric patients, but is very common (72%) in patients admitted after surgery for spinal fusion or craniofacial reconstruction. Patients who undergo surgery for correction of scoliosis and craniofacial anomalies should have serum magnesium levels monitored closely after surgery. In other patients, a neural network or multiple regression model could help predict which patients would be at risk of developing hypomagnesemia, thereby focusing testing on patients likely to benefit from such testing. PMID- 11057814 TI - Automated computerized intensive care unit severity of illness measure in the Department of Veterans Affairs: preliminary results. SISVistA Investigators. Scrutiny of ICU Severity Veterans Health Sysyems Technology Architecture. AB - OBJECTIVE: To evaluate the feasibility of an automated intensive care unit (ICU) risk adjustment tool (acronym: SISVistA) developed by selecting a subset of predictor variables from the Acute Physiology and Chronic Health Evaluation (APACHE) III available in the existing computerized database of the Department of Veterans Affairs (VA) healthcare system and modifying the APACHE diagnostic and comorbidity approach. DESIGN: Retrospective cohort study. SETTING: Six ICUs in three Ohio Veterans Affairs hospitals. PATIENT SELECTION: The first ICU admission of all patients from February 1996 through July 1997. OUTCOME MEASURE: Mortality at hospital discharge. METHODS: The predictor variables, including age, comorbidity, diagnosis, admission source (direct or transfer), and laboratory results (from the +/- 24-hr period surrounding admission), were extracted from computerized VA databases, and APACHE III weights were applied using customized software. The weights of all laboratory variables were added and treated as a single variable in the model. A logistic regression model was fitted to predict the outcome and the model was validated using a boot-strapping technique (1,000 repetitions). MAIN RESULTS: The analysis included all 4,651 eligible cases (442 deaths). The cohort was predominantly male (97.5%) and elderly (63.6 +/- 12.0 yrs). In multivariate analysis, significant predictors of hospital mortality included age (odds ratio [OR], 1.06; 95% confidence interval [CI], 1.04-1.09), comorbidity (OR, 1.11; 95% CI, 1.08-1.15), total laboratory score (OR, 1.07; 95% CI, 1.06-1.08), direct ICU admission (OR, 0.39; 95% CI, 0.31-0.49), and several broad ICU diagnostic categories. The SISVistA model had excellent discrimination and calibration (C statistic = 0.86, goodness-of-fit statistics; p > .20). The area under the receiver operating characteristic curve of the validated model was 0.86. CONCLUSIONS: Using common data elements often found in hospital computer systems, SISVistA predicts hospital mortality among patients in Ohio VA ICUs. This preliminary study supports the development of an automated ICU risk prediction system on a more diverse population. PMID- 11057815 TI - Ventilator circuit and secretion management strategies: a Franco-Canadian survey. AB - OBJECTIVE: To determine the use of ventilator circuit and secretion management strategies in France and Canada. DESIGN: Binational cross-sectional survey. POPULATION: Intensive care unit (ICU) directors in French and Canadian university hospitals. MEASUREMENTS: We compared responses between countries regarding the use of seven circuit and secretion strategies, the rationales against their use, decisional responsibility for these strategies, whether ventilator-associated pneumonia (VAP) practice was audited, and whether VAP prevention guidelines addressing these strategies were used. RESULTS: The response rate was 72/84 (85.7%) for French and 31/32 (96.9%) for Canadian ICUs. Endotracheal intubation was predominantly oral in both countries. Changing the ventilator circuits only for every new patient was more frequent in France than in Canada (p < .0001). Heated humidifiers were used more in Canada than France (p = .0003). Closed endotracheal suctioning was used more frequently in Canada (p < .0001). In both countries, subglottic secretion drainage and kinetic beds were rarely used. Semirecumbent positioning was reported more often by French than Canadian ICUs (p = .003). Reasons for nonuse of these strategies included adverse effects (heat and moisture exchangers), cost (kinetic beds), lack of convincing benefit (subglottic secretion drainage), and nurse inconvenience (semirecumbency). Decisional responsibility for each strategy differed among institutions. VAP prevention practice was periodically reviewed in 53% of French and 68% of Canadian ICUs (p = .20). VAP prevention guidelines were used in 64% and 30% of these ICUs, respectively (p = .002). CONCLUSIONS: Our study does not support the notion that published recommendations substantially impact reported use of several ventilator circuit and secretion management strategies. Based on the use of more frequent ventilator circuit changes, closed suctioning systems, heated humidifiers, and respiratory therapists, ventilator circuit and secretion management practice appears more costly in Canada than in France. PMID- 11057816 TI - Pulse oximeter-induced digital injury: frequency rate and possible causative factors. AB - OBJECTIVE: To study the frequency rate of and factors associated with pulse oximeter-induced digital injury (PIDI). DESIGN: Prospective descriptive study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: All patients with a length of stay of >2 days. INTERVENTIONS AND MAIN RESULTS: We monitored 125 patients by using a pulse oximeter with a nondisposable clip-on probe changed every 3-4 hrs from one finger to another. Daily inspection was performed with special attention to digital injury. Factors implicated in the pathogenesis of PIDI, such as vasopressor therapy, hypotension, hypoxia, hypothermia, and arterial cannulation of the radial or ulnar artery were recorded daily. A total of 22 patients received norepinephrine and dopamine, 34 patients were given dopamine, and 69 patients did not receive vasoactive drugs. PIDI developed in 6 of 125 patients, five in the norepinephrine/dopamine group, one in the dopamine group, and none in the patients not receiving vasopressors. When comparing the frequency rate of PIDI in patients receiving norepinephrine (5 of 22) with patients not receiving norepinephrine (1 of 103) the relationship between the use of norepinephrine and PIDI is significant (p < .001). However, this relationship may also be explained by the fact that patients in the norepinephrine group were more severely ill than patients not requiring norepinephrine were. This is reflected by a higher median severity of illness score (Simplified Acute Physiology Score II) (p = .001), median duration of hypotension (p < .001), median number of saturation drops (p < .001), and higher mortality rate (p < .001). Hypothermia did not occur in any of the patients. There was no significant difference between the median right-left difference in duration of arterial cannulation between the two subgroups (p = .8). In all surviving patients, PIDI healed without permanent damage. CONCLUSIONS: In the studied population of critically ill patients in a surgical intensive care unit, the frequency rate of PIDI was 5% (6 of 125). Although an association with the use of norepinephrine was found, this small number of cases does not allow definite statistical conclusions concerning a relationship between the possible causative factors and PIDI. However, severely ill patients, as indicated by their need for more aggressive vasopressors, are more likely to develop PIDI. In survivors, PIDI healed without sequelae. PMID- 11057817 TI - Severe heart failure secondary to 5-fluorouracil and low-doses of folinic acid: usefulness of an intra-aortic balloon pump. AB - OBJECTIVES: To report 5-fluorouracil in combination with folinic acid as a cause of severe nonischemic heart failure and to demonstrate the potential usefulness of an intra-aortic balloon pump. DESIGN: Case report. SETTING: An adult, 19-bed medical/surgical intensive care unit of a university hospital. PATIENTS: A patient, who developed severe heart failure secondary to 5-fluorouracil infusion with low-dose folinic acid, which was introduced to treat a rectal cancer, was transferred from a cancer institute to our intensive care unit 4 days after the treatment was initiated. INTERVENTIONS: Electrocardiography, determination of level of cardiac enzymes, echocardiography, radial arterial catheterization, mechanical ventilatory support, continuous venovenous hemodialysis, vasopressors, and secondary intra-aortic balloon pump. MEASUREMENT AND MAIN RESULTS: During shock, the patient's systolic blood pressure progressively decreased to 70 mm Hg, despite inotropic agents and vasopressors. Transesophageal echocardiography showed a calculated left ventricular ejection fraction within 20% with global hypokinesia. Electrocardiography showed sinus tachycardia with only nonspecific ST-T changes. Results of serial determination of levels of cardiac enzymes were not significant for myocardial infarction. Treatment with an intraaortic balloon pump was initiated and resulted in a dramatical improvement within 48 hrs. The patient was gradually weaned from vasopressors and the intra-aortic balloon pump. By the tenth day, echocardiography showed a septoapical hypokinesia with a 50% left ventricular ejection fraction. On the 30th day, the echocardiography was considered normal. CONCLUSION: Intravenous 5-fluorouracil in combination with low doses of folinic acid can induce severe nonischemic heart failure. In such a case, an intra-aortic balloon pump could be useful by providing left ventricular function support when inotropic agents and vasopressors fail to restore normal hemodynamics. PMID- 11057818 TI - Results of the Multidisciplinary Critical Care Knowledge Assessment Program, 2000 Exam. PMID- 11057819 TI - Surfactant: replacement, and more. PMID- 11057820 TI - Oxygen delivery in surgical patients--doesn't work, or does it? PMID- 11057821 TI - Percutaneous tracheostomy in patients without cervical spine clearance. PMID- 11057822 TI - Is the way to a man's heart really through his stomach? PMID- 11057823 TI - Cellular injury in sepsis. PMID- 11057824 TI - What if your hospital informatics department could provide a severity adjuster? PMID- 11057825 TI - Miracles and magic bullets: translating science into better care. PMID- 11057826 TI - Serum cholesterol and mortality in patients with multiple organ failure. PMID- 11057827 TI - Lack of effect of a ventilation strategy in preventing long-term lung injury. PMID- 11057828 TI - "Terminal" wean is the wrong term. PMID- 11057829 TI - Electrocardiographic guidance in placing central venous catheters. PMID- 11057830 TI - Effects of high hydrostatic pressures on living cells: a consequence of the properties of macromolecules and macromolecule-associated water. AB - Sixty percent of the Earth's biomass is found in the sea, at depths greater than 1000 m, i.e., at hydrostatic pressures higher than 100 atm. Still more surprising is the fact that living cells can reversibly withstand pressure shifts of 1000 atm. One explanation lies in the properties of cellular water. Water forms a very thin film around macromolecules, with a heterogeneous structure that is an image of the heterogeneity of the macromolecular surface. The density of water in contact with macromolecules reflects the physical properties of their different domains. Therefore, any macromolecular shape variations involving the reorganization of water and concomitant density changes are sensitive to pressure (Le Chatelier's principle). Most of the pressure-induced changes to macromolecules are reversible up to 2000 atm. Both the effects of pressure shifts on living cells and the characteristics of pressure-adapted species are opening new perspectives on fundamental problems such as regulation and adaptation. PMID- 11057831 TI - Role of Ca2+ in membrane excitation and cell motility in Characean cells as a model system. AB - The Characeae internodal cell is excitable and generates an action potential. The depolarizing phase of the action potential primarily reflects the activation of Cl- channels, which takes place in a Ca2+-dependent manner. Namely, an increase in the Ca2+ influx takes place at the very beginning of the action potential and heightens the Ca2+ concentration in the cytoplasm ([Ca2+]c). The high [Ca2+]c activates Cl- channels at the plasmalemma, resulting in a large depolarization. The high [Ca2+]c also acts as a signal to induce a tonoplast action potential and the instantaneous cessation of cytoplasmic streaming; the tonoplast action potential also is caused by Ca2+-induced activation of Cl- channels at the tonoplast, and the cessation is a result of inhibition of the actin-myosin interaction by Ca2+. When the cytoplasm of the Characeae cell, especially in Nitella flexilis, is hydrated rapidly, [Ca2+], also increases through Ca2+ release from an intracellular store(s). The release may be triggered by the stretching of endomembranes caused by osmotic swelling of the Ca2+ store lumens. Although the origin of Ca2+ is different from that in the case of an action potential, high [Ca2+]c not only induces membrane depolarization through activation of the Cl- channel in a Ca2+-dependent manner but also inhibits cytoplasmic streaming in Characeae. PMID- 11057832 TI - Calcium in ciliated protozoa: sources, regulation, and calcium-regulated cell functions. AB - In ciliates, a variety of processes are regulated by Ca2+, e.g., exocytosis, endocytosis, ciliary beat, cell contraction, and nuclear migration. Differential microdomain regulation may occur by activation of specific channels in different cell regions (e.g., voltage-dependent Ca2+ channels in cilia), by local, nonpropagated activation of subplasmalemmal Ca stores (alveolar sacs), by different sensitivity thresholds, and eventually by interplay with additional second messengers (cilia). During stimulus-secretion coupling, Ca2+ as the only known second messenger operates at approximately 5 microM, whereby mobilization from alveolar sacs is superimposed by "store-operated Ca2+ influx" (SOC), to drive exocytotic and endocytotic membrane fusion. (Content discharge requires binding of extracellular Ca2+ to some secretory proteins.) Ca2+ homeostasis is reestablished by binding to cytosolic Ca2+-binding proteins (e.g., calmodulin), by sequestration into mitochondria (perhaps by Ca2+ uniporter) and into endoplasmic reticulum and alveolar sacs (with a SERCA-type pump), and by extrusion via a plasmalemmal Ca2+ pump and a Na+/Ca2+ exchanger. Comparison of free vs total concentration, [Ca2+] vs [Ca], during activation, using time resolved fluorochrome analysis and X-ray microanalysis, respectively, reveals that altogether activation requires a calcium flux that is orders of magnitude larger than that expected from the [Ca2+] actually required for local activation. PMID- 11057833 TI - Mitogen-activated protein [MAP] kinase pathways in plants: versatile signaling tools. AB - Mitogen-activated protein kinases (MAPKs) are important signaling tools in all eukaryotes, and function in mediating an enormous variety of external signals to appropriate cellular responses. MAPK pathways have been studied extensively in yeast and mammalian cells, and a large body of knowledge on their functioning has accumulated, which is summarized briefly. Plant MAPK pathways have attracted increasing interest, resulting in the isolation of a large number of different components of MAPK cascades. Studies on the functions of these components have revealed that MAPKs play important roles in the response to a broad variety of stresses, as well as in the signaling of most plant hormones and in developmental processes. Finally, the involvement of various plant phosphatases in the inactivation of MAPKs is discussed. PMID- 11057834 TI - Why aren't we using beta blockers after acute MI? PMID- 11057835 TI - Family physicians and accutane. PMID- 11057836 TI - Urinary tract infections: 2000 update. PMID- 11057837 TI - Pneumocystis carinii pneumonia in HIV-infected patients. PMID- 11057838 TI - The AAP practice parameter on urinary tract infections in febrile infants and young children. American Academy of Pediatrics. AB - The Committee on Quality Improvement of the American Academy of Pediatrics developed an evidence-based practice parameter on the diagnosis, treatment and evaluation of the initial urinary tract infection in febrile infants and young children, two months to two years of age. The practice parameter consists of 11 recommendations and four areas for future research. Recommendations 1 through 5 address diagnosis and emphasize the need for culturing appropriately collected specimens; diagnosis should not be based on urinalysis or on culture of specimens from urine-collection bags. Recommendations 6 through 10 address treatment, identifying trimethoprim-sulfamethoxazole as superior to amoxicillin and establishing the duration of treatment as seven to 14 days. Children should receive antimicrobial coverage until imaging studies have been completed. Recommendation 11 addresses imaging; all infants should undergo ultrasonography, and a lower tract study is strongly encouraged as well. Future research should quantitatively address the relationship between renal scarring in childhood and renal impairment and hypertension in adults. Less invasive methods of diagnosis and imaging are highly desirable and should be developed. PMID- 11057839 TI - Use of systemic agents in the treatment of acne vulgaris. AB - Effective treatment of acne vulgaris can prevent emotional and physical scarring. Therapy varies according to the severity of the disease. Topical medication is generally adequate in clearing comedonal acne, while inflammatory acne usually requires the addition of oral medication. Systemic antibiotics are used most frequently and can be highly effective. Newer formulations of combined oral contraceptives are also helpful in modulating sebum production in the female patient. Severe nodulocystic acne that does not respond to topical retinoids and systemic antibiotics may be treated with isotretinoin. However, the side effect profile of this medication is extensive, and physicians should be well-versed in its potential adverse effects. PMID- 11057840 TI - Using progestins in clinical practice. AB - Progestational agents have many important functions, including regulation of the menstrual cycle, treatment of dysfunctional uterine bleeding, prevention of endometrial cancer and hyperplastic precursor lesions, and contraception. Because of the reported side effects of synthetic analogs called "progestins," there has been interest in replicating the natural hormone for clinical use. Natural progesterone is obtained primarily from plant sources and is currently available in injectable, intravaginal and oral formulations. An oral micronized progesterone preparation has improved bioavailability and fewer reported side effects compared with synthetic progestins. Adolescents and perimenopausal women may require progestational agents for the treatment of dysfunctional uterine bleeding resulting from anovulatory cycles. These agents may also be used in women at risk for endometrial hyperplasia because of chronic unopposed estrogen stimulation. Progestin-only contraceptives can be used in women with contraindications to estrogen; however, efficacy requires rigorous compliance. New progestins for use in combination oral contraceptive pills were specifically developed to reduce androgenic symptoms. It is unclear whether these progestins increase the risk of venous thromboembolic disease. Progestin-only emergency contraception offers a regimen that is more effective than combination oral contraceptive pills, with fewer reported side effects. PMID- 11057841 TI - Optimizing beta-blocker use after myocardial infarction. AB - Although beta-adrenergic blockers can significantly reduce mortality after a myocardial infarction, these agents are prescribed to only a minority of patients. Underutilization of beta blockers may be attributed, in part, to fear of adverse effects, especially in the elderly and in patients with concomitant disorders such as diabetes or heart failure. However, studies have shown that such patients are precisely the ones who derive the greatest benefit from beta blockade. Advancing age or the presence of potentially complicating disease states is usually not a justification for withholding beta-blocker therapy. With use of cardioselective agents and through careful dosing and monitoring, the benefits of beta blockers after myocardial infarction far outweigh the potential risks in most patients. PMID- 11057842 TI - Topical fluoroquinolones for eye and ear. AB - Topical fluoroquinolones are now available for use in the eye and ear. Their broad spectrum of activity includes the common eye and ear pathogens Staphylococcus aureus and Pseudomonas aeruginosa. For the treatment of acute otitis externa, these agents are as effective as previously available otic preparations. For the treatment of otitis media with tympanic membrane perforation, topical fluoroquinolones are effective and safe. These preparations are approved for use in children, and lack of ototoxicity permits prolonged administration when necessary. Topical fluoroquinolones are not appropriate for the treatment of uncomplicated conjunctivitis where narrower spectrum agents suffice; they represent a simplified regimen for the treatment of bacterial keratitis (corneal ulcers). When administered topically, fluoroquinolones are well tolerated and offer convenient dosing schedules. Currently, bacterial resistance appears limited. PMID- 11057843 TI - Guidelines on migraine: Part 2. General principles of drug therapy. PMID- 11057844 TI - Deficiency causing mutations and common polymorphisms in the factor XIII-A gene. PMID- 11057845 TI - Thrombotic thrombocytopenic purpura: a paradigm shift? PMID- 11057846 TI - Molecular characterization of a multiethnic group of 21 patients with type 3 von Willebrand disease. AB - Type 3 von Willebrand disease is a rare autosomal disorder characterized by unmeasurable levels of von Willebrand factor and severe hemorrhagic symptoms. We studied a multiethnic group of 37 patients, from Italy (n = 14), Iran (n = 10) and India(n = 13) to identify the molecular defects and to evaluate genetic heterogeneity among these populations. Twenty-one patients (6 Italians, 9 Iranians and 6 Indians) were fully characterized at the molecular level. Twenty four different gene alterations were identified, 20 of which have not been described previously. The majority of the mutations caused null alleles, 11 being nonsense mutations (Q218*, W222*, R365*, R373*, E644*, Q706*, S1338*, Q1346*, Y1542*, R1659*, E2129*), 4 small deletions (437delG, 2680delC, 6431delT, del 8491 8499), 3 possible splice site mutations [IVS9(-1)g-->a, IVS29(+10)c-->t, IVS40( 1)g-->c], 3 candidate missense mutations (C275S, C2174G, C2804Y), 2 small insertions (7375insC, 7921insC) and 1 large gene deletion. The latter mutation was associated with the development of alloantibodies to VWF, but this complication was also found in a patient homozygous for a nonsense mutation (Q1346*). Due to the ethnic origin of the patients most of them were the offspring of consanguineous marriages and so were homozygous for the mutations found (18/21). Our results indicate that molecular defects responsible for type 3 VWD are scattered throughout the entire VWF gene (from exon 3 to 52), and that there is no prevalent and common gene defect in the three populations studied by us. PMID- 11057847 TI - Discrimination of von Willebrands disease (VWD) subtypes: direct comparison of von Willebrand factor:collagen binding assay (VWF:CBA) with monoclonal antibody (MAB) based VWF-capture systems. AB - Discrimination of von Willebrand's Disease (VWD) subtypes is important since it influences management. Qualitative [ie Type 2A, 2B, 2M] defects exhibit von Willebrand factor (VWF) discordance and give high VWF:Ag to VWF:'activity' ratios. Classically, VWF:'activity' is assessed using the VWF:RCof assay. The VWF:CBA is an ELISA-based VWF-functional adhesive assay which has consistently proved to be superior to VWF:RCof. A commercially available monoclonal antibody (MAB) based ELISA assay system claimed to mimic a VWF:RCof-like activity has also been recently described ('SE'), as has the production and characterisation of a large number [n = 10] of locally generated anti-VWF MAB. In the current study, we have adapted these MAB to in-house ELISA assays to assess their utility for VWD diagnosis and subtype discrimination, and to compare them with other assay systems. Thus, the VWF:CBA, VWF:RCof by agglutination, the SE assay, and in-house MAB based assays have been directly compared for their ability to discriminate Type 1 [n = 9] from Type 2 VWD samples [phenotypes 2A and 2B; n = 11]. In summary, MAB-based systems can be used to measure VWF and confirm a diagnosis of VWD, as well as exhibiting some VWD-subtype-discriminatory capabilities. However, better evidence of VWF-discordance was usually achieved using the VWF:RCof (agglutination) assay, while the greatest degree of VWF-discordance was consistently observed using the VWF:CBA assay. In conclusion, the VWF:CBA assay proved to offer the best diagnostic predictive tool for a Type 2 VWD defect, while MAB-based systems appear to be less effective in this regard. PMID- 11057848 TI - Predicting adverse outcome in patients with acute pulmonary embolism: a risk score. AB - Reliable prediction of adverse outcomes in acute pulmonary embolism may help choose between in-hospital and ambulatory treatment. We aimed to identify predictors of adverse events in patients with pulmonary embolism and to generate a simple risk score for use in clinical settings. We prospectively followed 296 consecutive patients with pulmonary embolism admitted through the emergency ward. Logistic regression was used to predict death, recurrent thromboembolic event, or major bleeding at 3 months. Thirty patients (10.1%) had one or more adverse events during the 3-month follow-up period: 25 patients (8.4%) died, thromboembolic events recurred in 10 patients (3.4%), and major bleeding occurred in 5 patients (1.7%). Factors associated with an adverse outcome in multivariate analysis were cancer, heart failure, previous deep vein thrombosis, systolic blood pressure <100 mmHg, arterial PaO2 <8 kPa, and presence of deep vein thrombosis on ultrasound. A risk score was calculated by adding 2 points for cancer and hypotension, and 1 point each for the other predictors. A score of 2 best identified patients at risk of an adverse outcome in a receiver operating characteristic curve analysis. Of 180 low-risk patients (67.2%) (score < or =2), only 4 experienced an adverse outcome (2.2%), compared to 23 (26.1%) of 88 high risk patients (score > or =3). A simple risk score based on easily available variables can accurately identify patients with pulmonary embolism at low risk of an adverse outcome. Such a score may be useful for selecting patients with pulmonary embolism eligible for outpatient care. PMID- 11057849 TI - Blood rheology, cardiovascular risk factors, and cardiovascular disease: the West of Scotland Coronary Prevention Study. AB - The West of Scotland Coronary Prevention Study (WOSCOPS) showed that pravastatin reduced the risk of coronary heart disease (CHD) events in 6,595 middle-aged hypercholesterolaemic men aged 45-64 years without prior myocardial infarction followed for an average of 4.9 years. We hypothesised prospectively (a) that baseline levels of haemorheological variables were related to baseline and incident CHD and to mortality; and (b) that reduction in lipoproteins by pravastatin would lower plasma and blood viscosity, a potential contributory mechanism to CHD events. We therefore studied plasma and blood viscosity, fibrinogen, haematocrit, and blood cell counts at baseline and 1 year. At baseline, plasma and blood viscosity were related to risk factors, CHD measures, and claudication. On univariate analysis, baseline levels of all rheological variables (except platelet count) were related to incident CHD; CHD mortality; and total mortality. On multivariate analysis including baseline CHD and risk factors, plasma and blood viscosity, haematocrit and white cell count each remained significantly associated with incident CHD; while fibrinogen remained an independent predictor of mortality (all p < 0.03). After one year, lipoprotein reduction by pravastatin was associated with significant reductions (about one quarter of a standard deviation) in plasma viscosity (mean difference 0.02 mPa.s, p <0.001) and in blood viscosity (mean difference 0.06 mPa.s, p<0.001), but was not associated with significant changes in other rheological variables. We therefore suggest that pravastatin therapy, which reduces elevated lipoproteins in hypercholesterolaemic men, may lower risks of CHD and mortality partly by lowering plasma and blood viscosity. Further studies are required to test this hypothesis. PMID- 11057850 TI - Low molecular weight heparin (enoxaparin) versus oral anticoagulant therapy (acenocoumarol) in the long-term treatment of deep venous thrombosis in the elderly: a randomized trial. AB - This study aims to establish the relative effectiveness and safety of low molecular weight heparin in elderly patients with venous thrombosis in order to find an alternative to oral anticoagulant therapy with less bleeding complications in the long-term treatment of deep venous thrombosis (DVT). One hundred consecutive elderly patients (>75 years old) with venographically demonstrated proximal DVT were included in a randomized trial. All patients were treated for ten days with adjusted doses of intravenous heparin. Informed consent was obtained and on the eight day, patients were randomly allocated to receive acenocoumarol (INR 2.0-3.0) or subcutaneous enoxaparin (4000 anti-Xa units once a day) for three months. All patients were followed-up clinically and venographically for a one year period. The results were analyzed with Fisher's exact test or chi-square test as appropriate. During the treatment and surveillance period, 6 of the 50 patients (12%) who received acenocoumarol and 8 of the 50 patients (16%) who received enoxaparin had new episodes of venous thromboembolism confirmed by objective testing (p = 0.6; 95% CI for the difference: -19.5 to 11.5). Hemorrhagic complications occurred in six of the 50 patients (12%) who received acenocoumarol and in one (2%) of those on enoxaparin (p = 0.1; 95% CI for the difference: -1.8 to 21.8). Vertebral fractures developed in 2 patients (4%) in the enoxaparin group (p = 0.5; 95% CI for the difference: 11.4 to 3.4). These results show that fixed dose enoxaparin seems to be effective and safe in the long-term treatment of proximal DVT in the elderly. In comparison with oral anticoagulants, the findings are inconclusive due to the wide confidence intervals for differences between outcomes, however they suggest that the former may have less bleeding complications with similar efficacy. PMID- 11057851 TI - Increased fibrinogen levels and hemostatic abnormalities in patients with arteriolar nephrosclerosis: association with cardiovascular events. AB - Increased plasma fibrinogen levels and hemostatic abnormalities suggestive of a prothrombotic state are present in patients with end-stage renal failure and could contribute to increased cardiovascular morbidity in these patients. We investigated the relationship between abnormalities of the hemostatic system and the degree of renal failure and whether these abnormalities are associated with increased prevalence of cardiovascular events in patients with arteriolar nephrosclerosis. In 425 patients recruited at a hypertension clinic we assessed the renal function by creatinine clearance, urinary protein excretion, and microalbuminuria, the prevalence of atherosclerotic disease, and measured prothrombin time, activated partial thromboplastin time. fibrinogen, prothrombin fragment 1+2 (F1+2), D-dimer, and antithrombin. Early impairment of renal function (creatinine clearance, 30 to 89 ml/min per 1.73 m2 of body surface area) caused by arteriolar nephrosclerosis was found in 172 patients. Patients with early renal failure were significantly older and had significantly greater values of blood pressure, plasma fibrinogen, F1+2, and D-dimer than patients with normal renal function. Elevated D-dimer and fibrinogen levels were independently associated with the presence of decreased creatinine clearance. Log fibrinogen, log F1+2, and log D-dimer were inversely correlated with creatinine clearance. The prevalence of coronary artery, cerebrovascular, and peripheral vascular disease was significantly greater in patients with mild renal failure than in those with normal renal function. Elevated levels of fibrinogen and D-dimer were associated with the presence of atherosclerotic disease independent of renal function and other risk factors. In conclusion, changes in hemostatic parameters occur early in the course of renal failure in patients with arteriolar nephrosclerosis, suggesting a prothrombotic state that may contribute to the risk for atherosclerotic disease at all levels of renal function. PMID- 11057852 TI - Insulin sensitivity and hemostatic factors in clinically healthy 58-year-old men. AB - The objective of this cross-sectional study was to investigate the relationship between factors of the coagulation- and fibrinolysis systems and insulin sensitivity in 104 clinically healthy, 58-years-old men. Insulin sensitivity (hyperinsulinemic euglycemic clamp) adjusted for lean body mass, the metabolic syndrome according to a suggested definition, and different factors in the coagulation- and fibrinolysis system were determined. Subjects with the metabolic syndrome were characterised by increases in PAI-1 activity, tPA antigen, protein C and protein S and low concentrations of tPA activity. Insulin sensitivity was independently and reversibly associated with PAI-1 (p = 0.014) and directly with tPA activity (p = 0.001). Insulin sensitivity was also significantly negatively associated with protein S and protein C and several components in the metabolic syndrome, however not remaining significant in multivariate analyses. Protein C and protein S were significantly associated with PAI-1 activity, tPA activity (negatively), tPA antigen and antithrombin III. In conclusion, the data indicated that insulin resistance and several of the clustering components in the metabolic syndrome are accompanied by increased plasma concentrations of the anticoagulatory proteins C and S which may represent a mechanism which counteracts the concomitantly occurring hypofibrinolysis. PMID- 11057853 TI - Contribution of the cystathionine beta-synthase gene (844ins68) polymorphism to the risk of early-onset venous and arterial occlusive disease and of fasting hyperhomocysteinemia. AB - The frequency of the heterozygous 844ins68 mutation of the cystathionine beta synthase (CBS) gene and of its association with the homozygous C677T transition of the methylenetetrahydrofolate reductase (MTHFR) gene, plasma fasting tHcy, folate and vitamin B12 levels were evaluated in 309 consecutive patients with objectively diagnosed early-onset venous (n = 200) or arterial thromboembolic disease (n = 109) recruited over 25 months in Milan (North Italy) and Naples (South Italy). The above gene polymorphisms were also evaluated in a population of 787 unmatched controls, 204 of whom--similar to patients for age- and sex distribution--had fasting tHcy, vitamins and activated protein C resistance measured in their plasma. Moderate fasting hyperhomocysteinemia was detected in 15.5% of patients and in 5.9% of 204 controls (Mantel-Haenszel OR after stratification for type of occlusive disease and gender: 2.88; 1.48-5.32). The frequencies of the 677TT mutation of the MTHFR gene and of the heterozygous 844ins68 insertion of the CBS gene were not significantly different in the patient (19.4% and 6.9%) and the control population (16.5% and 7.8%), but the association of the two gene polymorphisms found in 3.9% of patients and in 1.1% of controls - was significantly associated with an increased risk of venous or arterial occlusive diseases (RR = 3.63; 1.48-8.91). The MTHFR 677TT mutation (RR: 6.92; 3.86-12.4) and its association with the 844ins68 insertion (RR: 21.9; 8.35 57.4), but not the isolated insertion (RR: 0.71), were more frequent in patients and controls with fasting hyperhomocysteinemia than in normohomocysteinemic subjects, irrespective of the type of occlusive disease (venous or arterial). When adjusted for determinants of hyperhomocysteinemia in the patient and the control populations (generalized linear model), fasting tHcy levels were significantly higher in subjects with association of the two gene abnormalities (24.2+/-3.8 micromol/L) than in subjects with the MTHFR 677TT mutation only (14.0+/-5.8 micromol/L, p = 0.004). Activated protein C resistance was significantly more prevalent in venous patients (9.9%) than in controls (3.9%, OR = 2.69; 1.08-6.88). Six of 21 venous patients with APC-resistance also had hyperhomocysteinemia (RR = 5.04; 0.68-37.6), but isolated fasting hyperhomocysteinemia retained statistical significance for the association with venous occlusive disease (RR = 2.84; 1.34-6.01). Heterozygosity for the 844ins68 mutation of the CBS gene is not per se a risk factor for premature arterial and/or venous occlusive diseases. However, when detected in combination with thermolabile MTHFR, it increases by almost 4-fold the risk of occlusive diseases (arterial and/or venous), by increasing the risk and the degree of fasting hyperhomocysteinemia. PMID- 11057854 TI - Antithrombin therapy for severe preeclampsia: results of a double-blind, randomized, placebo-controlled trial. BI51.017 Study Group. AB - A double-blind, randomized, placebo-controlled trial was conducted to evaluate whether treatment with Antithrombin (AT) concentrates improved the clinical and perinatal outcome in patients with severe preeclampsia. Severe preeclamptic patients (24 to 35 weeks of gestation. Gestosis Index (GI) > or = 6 points) were randomized into two groups: 66 received AT and 67 received placebo. There were no statistical differences in the clinical profiles of the two groups. Study drugs were given intravenously once daily for 7 consecutive days. Maternal symptoms were evaluated from the difference of GI between before and after treatment, and fetal findings were evaluated from the changes of the biophysical profile score and the estimated fetal weight gain. Improvement was significantly greater in the AT group for both the GI (p = 0.020) and the estimated fetal weight gain (p = 0.029). The improvement of coagulation parameters was also evaluated. The D-dimer levels increased significantly in the placebo group (p = 0.026), but did not change in the AT group. Gestation was significantly prolonged (p = 0.007), and the number of low-birth weight infants was significantly smaller (p = 0.011) in the AT group. No adverse events related to AT were observed. It is revealed that AT concentrate therapy for preeclampsia is effective and safe, leading to an improved perinatal outcome. PMID- 11057855 TI - Factor XIII deficiency causing mutation, Ser295Arg, in exon 7 of the factor XIIIA gene. AB - Inherited factor XIII (FXIII) deficiency is an autosomal recessive disorder which results in a serious bleeding diathesis, problems with wound healing and a very high risk of recurrent miscarriage in deficient females. We have analysed the molecular basis of factor XIII deficiency in two patients and their parents, who originate from the North of Pakistan. Four sequence changes were identified: an AGC-->AGG (Ser-->Arg) FXIII deficiency-causing mutation in codon 295; G-->A at position -246 upstream of exon 1; T-->C and C-->T at positions -23 and -24, respectively, in intron 9. Using molecular modelling we predict that the Ser295Arg mutation would prevent the FXIIIA molecule from folding correctly and thus result in an unstable FXIIIA mutant polypeptide. The sequence changes -246G- >A, -23T-->C and -24C-->T are normal polymorphisms. RT-PCR analysis demonstrates that the intronic sequence changes do not appear to affect the accuracy of FXIIIA RNA processing. PMID- 11057856 TI - Effect of Val34Leu polymorphism on the activation of the coagulation factor XIII A. AB - Coagulation factor XIII (FXIII) is a protransglutaminase involved in the last step of the coagulation cascade by stabilising the fibrin clot. Recently, a common variation (FXIII Val34Leu) has been associated with a decreased risk of myocardial infarction and deep venous thrombosis. Val34Leu is critically located near the thrombin activation site of FXIII-A. In this study we investigated its effects on the activation of FXIII. Both recombinant and platelet-derived FXIII Val34Leu variants were shown to be more susceptible to thrombin cleavage than the wild type FXIII. The rate of enzymatic activation of FXIII Val34Leu was found increased, however, the specific activity of fully activated wild type FXIII and the Val34Leu mutant did not differ. During the course of thrombin-induced activation of FXIII fibrin gamma-chain dimerisation and alpha-chain polymerisation developed more rapidly with the Val34Leu mutant. The increased rate of fibrin stabilisation brought about by the Val34Leu FXIII seems to be paradoxically associated with a protective effect against pathological thrombosis. PMID- 11057857 TI - Ethnic heterogeneity of the factor XIII Val34Leu polymorphism. AB - A polymorphism in the coagulation factor XIII gene (FXIII Val34Leu) has been recently described to confer protection for arterial and venous thrombosis and to predispose to intracerebral hemorrhage. At present it is known that FXIII Val34Leu is prevalent in Caucasians, but information upon its distribution in different ethnic groups is scarce. We investigated the prevalence of FXIIIVal34Leu in 450 unrelated subjects of four ethnic groups: 97 Caucasians (Brazilians of European descent and Portuguese), 149 Blacks (Brazilians, and Africans from Cameroon, Zaire and Angola), 40 Asians (Japanese descendents) and 164 Amerindians from South America. PCR amplification of exon 2 of FXIII gene followed by MseI restriction-digestion was employed to define the genotypes. FXIIIVal34Leu was detected in 44.3% of the Caucasians, in 28.9% of the Blacks, in 2.5% of the Asians and in 51.2% of the Amerindians. These data confirm that FXIII Val34Leu is highly prevalent in Caucasians and indicate that it is rarer in populations of African origin. The very high frequency among Amerindians indicates that FXIII Val34Leu is not absent among Asians, and since it has a very low prevalence in Japanese, a heterogeneity in its distribution in Asia may be inferred. Taken together, our data showed that FXIII Val34Leu exhibits a significant ethnic heterogeneity, a finding that is relevant for studies relating this polymorphism with thrombotic and bleeding disorders. PMID- 11057858 TI - Characterization of cleaved plasma protein S with a monoclonal antibody-based assay. AB - A monoclonal antibody (mAb 5A5G2) recognized cleaved plasma protein S (PS) but not uncleaved PS. Interestingly, mAb 5A5G2 did not recognize thrombin-cleaved recombinant PS. Microsequencing of cleaved plasma PS showed a Q-S-T-N amino terminal sequence, inferring cleavage after the Arg 60 residue. The mAb epitope was located within the sequence encompassing residues 61 to 73, i.e. the carboxy terminal part of the thrombin-sensitive region (TSR). We used this mAb to develop an ELISA assay to quantify in vivo cleaved PS. In plasma from 10 normal subjects, about 10% of PS was cleaved (7.1% to 15.4%), with a more than 2-fold increase in the corresponding sera. We found increased levels of cleaved PS in 8 patients with disseminated intravascular coagulation (DIC) and decreased levels in 22 patients on long-term oral anticoagulant therapy, whereas cleaved PS levels were similar in 8 hemophiliacs and the 10 normal subjects. Cleaved PS levels did not correlate with prothrombin fragment 1+2 levels released after cleavage by FXa in any of the groups, suggesting that circulating FXa is not the main factor involved in the production of cleaved PS in vivo. PMID- 11057859 TI - Pre-clinical pharmacological profile of the novel glycoconjugate Org 36764 with both factor Xa and thrombin (IIa) inhibitory activities. AB - Org 36764, is an antithrombin III (AT) and thrombin binding carbohydrate, which accelerates the inactivation of both factor Xa and thrombin by AT. It displays in buffer an anti-Xa and anti-thrombin activity of 415 and 2 U/mg, respectively, compared to 172 and 114 U/mg, respectively, for unfractionated heparin (UFH), Org 36764 does not cross-react with HIT (heparin induced thrombocytopenia) antibodies and is not neutralised by PF4. In experimental models in rats, on a molar basis. Org 36764 was more active than the pentasaccharide SanOrg 34006 (= AT binding domain of Org 36764) in arterial thrombosis, but both were equally active in venous thrombosis. In arterial thrombosis following endothelial damage by ferric chloride, Org 36764 was more active than the LMW heparin enoxaparin and SanOrg 34006 and similar active to UFH. At AT saturating doses the bleeding enhancement was not more than 3.5 times the control value. Org 36764 was more active in suppressing in vivo thrombus formation on stents than UFH. SanOrg 34006 or a combination of ticlopidine and aspirin. The results indicate that the novel drug Org 36764 is a drug with antithrombotic potential against venous and arterial thrombosis. PMID- 11057860 TI - Sequence alignment between vWF and peptides inhibiting the vWF-collagen interaction does not result in the identification of a collagen-binding site in vWF. AB - We previously found that two peptides (N- and Q-peptide) selected by phage display for binding to an anti-vWF antibody, were able to inhibit vWF-binding to collagen (1). The sequence of those peptides could be aligned with the sequence in vWF at position 1129-1136 just outside the A3-domain. As the peptides represent an epitope or mimotope of vWF for binding to collagen we next wanted to study whether the alignment resulted in the identification of a new collagen binding site in vWF. We mutated the 1129-1136 VWTLPDQC sequence in vWF to VATAPAAC. Expressing this mutant vWF (7.8-vWF) in a fur-BHK cell line resulted in well processed 7.8-vWF containing a normal distribution of molecular weight multimers. However, binding studies of this mutant vWF to rat tail, human and calf skin collagens type 1, to human collagen types III and VI, revealed no decrease in vWF-binding to any of these collagens. Thus, although the N- and Q peptides did inhibit the vWF-collagen interaction, the resulting alignment with the vWF sequence did not identify a collagen binding site, pointing out that alignments (although with a high percentage of identity) do not always result in identification of binding epitopes. However, suprisingly removal of the A3-domain or changing the vWF sequence at position 1129-1136 resulted in an increase of vWF binding to human collagen type V1 and to rat tail collagen type 1, implying that these changes result in a different conformation of vWF with an increased binding to these collagens as a consequence. PMID- 11057861 TI - Hemophilia B with mutations at glycine-48 of factor IX exhibited delayed activation by the factor VIIa-tissue factor complex. AB - Gly-48 is in the conserved DGDQC sequence (residues 47-51 of human factor IX) of the first EGF (EGF-1)-like domain of factor IX. The importance of the Gly-48 is manifested by two hemophilia B patients; factor IXTainan and factor IXMalmo27, with Gly-48 replaced by arginine (designated IXG48R) and valine (IXG48V), respectively. Both patients were CRM+ exhibiting mild hemophilic episodes with 25% (former) and 19% (latter) normal clotting activities. We characterize both factor IX variants to show the roles of Gly-48 and the conservation of the DGDQC sequence in factor IX. Purified plasma and recombinant factor IX variants exhibited approximately 26%-27% normal factor IX's clotting activities with G48R or G48V mutation. Both variants depicted normal quenching of the intrinsic fluorescence by increasing concentrations of calcium ions and Tb3+, indicating that arginine and valine substitution for Gly-48 did not perturb the calcium site in the EGF-1 domain. Activation of both mutants by factor XIa appeared normal. The reduced clotting activity of factors IXG48R and IXG48V was attributed to the failure of both mutants to cleavage factor X: in the presence of only phospholipids and calcium ions, both mutants showed a 4 to approximately 7-fold elevation in Km, and by adding factor VIIIa to the system, although factor VIIIa potentiated the activation of factor X by the mutants factor IXaG48R and factor IXaG48V, a 2 to approximately 3-fold decrease in the catalytic function was observed with the mutant factor IXa's, despite that they bound factor VIIIa on the phospholipid vesicles with only slightly reduced affinity when compared to wild-type factor IXa. The apparent Kd for factor VIIIa binding was 0.83 nM for normal factor IXa, 1.74 nM for IXaG48R and 1.4 nM for IXaG48V. Strikingly, when interaction with the factor VIIa-TF complex was examined, both mutations were barely activated by the VIIa-TF complex and they also showed abnormal interaction with VIIa-TF in bovine thromboplastin-based PT assays. Taken together, our results suggest that mutations at Gly-48 altered the interaction of factor IX with its extrinsic pathway activator (VIIa-TF complex), its macromolecular substrate (factor X), and its cofactor (factor VIIIa). PMID- 11057862 TI - Homozygous 2bp deletion in the human factor VII gene: a non-lethal mutation that is associated with a complete absence of circulating factor VII. AB - We report the case of a 5-year-old boy with severe factor VII deficiency. The affected child presented at the age of 8 months and again at 18 months with bleeding from the gastrointestinal tract but the diagnosis of factor VII deficiency was not made until the age of 3 years. He was treated with fresh frozen plasma and subsequently factor VII concentrates and to date remains well. To identify the causative mutation, the factor VII gene was screened by SSCP and direct sequence analysis. A single homozygous 2 bp deletion (-CT) mutation was identified in exon 1a removing nucleotides 27/28 (codons 52/53). Both parents, who were first cousins, were heterozygous for the mutation. The mutation located in the prepropeptide of factor VII, results in a complete absence of factor VII in plasma. This case indicates that a complete absence of plasma factor VII is not necessarily a lethal condition. PMID- 11057863 TI - The effect of DDAVP infusion on thrombin generation in platelet-rich plasma of von Willebrand type 1 and in mild haemophilia A patients. AB - In von Willebrand disease (vWD) type 1 and mild haemophilia A patients we studied the effect of an infusion of DDAVP (0.3 microg/kg body weight) on thrombin generation in platelet-rich plasma (PRP) and platelet-poor plasma (PPP). Baseline thrombin generation in PRP was diminished both in the haemophilia A and vWD patients. It was normal in vWD plasma when sufficient procoagulant phospholipids were present, either via adding phospholipid vesicles to PPP or via scrambling of the platelet membrane with ionomycin in PRP. In haemophilia A plasma, thrombin generation did not normalize by providing procoagulant phospholipids. Treatment with DDAVP temporarily restored thrombin generation in PRP to normal in both diseases. To investigate the individual roles of von Willebrand factor (vWF) and factor VIII, we also studied the effect of factor VIII infusion on thrombin generation in a severe haemophilia patient. It appears that at a fixed normal vWF concentration, <25% factor VIII is sufficient for normal thrombin generation in PRP. At a sufficient factor VIII concentration, however, thrombin generation is still lower than normal in vWD patients; approximately 40% of vWF is required for half-normal thrombin generation in PRP. It thus appears that vWF is also a clotting factor, in the sense that it is required for normal thrombin generation. This underlines the importance of the interaction between coagulation and the platelets in normal haemostasis. Thrombin generation in PRP appears to be a suitable test to reflect the combined function. PMID- 11057864 TI - Sensitization of CD4+ T cells to coagulation factor VIII: response in congenital and acquired hemophilia patients and in healthy subjects. AB - Antibodies (Ab) that inhibit factor VIII (fVIII) may develop in patients with hemophilia A and rarely in individuals without congenital fVIII deficiency (acquired hemophilia). Synthesis of fVIII inhibitors requires CD4+ T cells. We investigated the proliferative response of blood CD4+ cells from 11 patients with congenital or acquired hemophilia and 12 healthy subjects, to recombinant human fVIII, and to pools of overlapping synthetic peptides spanning the sequences of individual fVIII domains. All patients had CD4+ cells that responded to fVIII. The intensity of the responses fluctuated over time: several patients had brief periods when they did not respond to fVIII. All healthy subjects had transient CD4+ responses to fVIII, that were significantly lower than those of hemophilia patients. Also, healthy subjects responded to fVIII less frequently and for shorter periods than hemophilia patients. All patients and healthy subjects recognized several fVIII domains: the A3 domain was recognized most strongly and frequently. The transient sensitization of CD4+ cells to fVIII in healthy subjects suggests that inadequate tolerization of CD4+ cells to fVIII, due to lack of endogenous fVIII, is an important factor in the development of clinically significant anti-fVIII antibodies in hemophilia A. PMID- 11057865 TI - Antibodies to beta2-glycoprotein I associated with antiphospholipid syndrome suppress the inhibitory activity of tissue factor pathway inhibitor. AB - Anionic phospholipid membranes have a dual role in blood coagulation: they are essential for the initiation and propagation as well as for the limitation and termination of the blood coagulation process. Patients with the anti-phospholipid syndrome (APS) carrying antibodies against complexes of anionic phospholipids and plasma proteins, show in vitro inhibited phospholipid dependent coagulation reactions, whereas in vivo the presence of these antibodies is associated with an increased risk of thrombosis. In this study we focussed on the effects of these anti-phospholipid antibodies on the regulation of TF-mediated factor Xa (FXa) generation in plasma. We hypothesized that anti-phospholipid antibodies interfere with the phospholipid-dependent inhibition by tissue factor pathway inhibitor (TFPI) of TF-induced coagulation. Indeed, total-IgG, anti-cardiolipin-IgG (aCL) and anti-beta2GPI-IgG, isolated from patient plasmas, all stimulated TF-induced FXa generation in normal plasma. This enhanced FXa generation was not observed when the patient's IgG was depleted of anti-beta2GPI-IgG or when normal plasma was depleted of beta2PGPI or TFPI. Taken together, these data indicate that antibodies to beta2GPI, circulating in patients with APS, suppress TFPI-dependent inhibition of TF-induced coagulation, which results in an increased FXa generation. PMID- 11057866 TI - Tissue factor encryption/de-encryption is not altered in the absence of the cytoplasmic domain. AB - Since the cytoplasmic domain of tissue factor (TF) appears to have a role in TF function beyond coagulation, experiments were conducted to determine whether the cytoplasmic domain also has a role in regulating procoagulant activity of TF present in the cell membrane. TF encryption was quantitated in human YU-SITI, U87 MG, and mouse 3T3 cells which were transfected for expression of human tissue factor or a construct lacking the cytoplasmic domain (TF(CD)). Comparison of intact cells (encrypted) with fully disrupted cells (de-encrypted) showed that TF and TF(CD) were equally encrypted with respect to function in fX activation. Moreover, cells expressing TF and TF(CD) were indistinguishable in their procoagulant responses to A23187-calcium and varied concentrations of nonionic detergents. TF in membrane vesicles spontaneously shed by U87-MG cells was largely, but incompletely, de-encrypted, and the degree of de-encryption was independent of the cytoplasmic domain. We conclude that the predominant mechanism(s) for encrypting TF procoagulant activity is independent of the cytoplasmic domain. PMID- 11057867 TI - A comparison of two sodium citrate concentrations in two evacuated blood collection systems for prothrombin time and ISI determination. AB - The prothrombin time is usually measured in citrated plasma. The W.H.O. recommended concentration of sodium citrate for blood collection for laboratory control of oral anticoagulant therapy is 0.109 M. Some evacuated blood collection systems include 0.105 M sodium citrate. The purpose of the present study was to establish the difference in ISI calibration between 0.109 and 0.105 M citrate, using 7 types of thromboplastin and various types of instrumentation. The two citrate concentrations were provided in both evacuated siliconised glass tubes and in evacuated polyethylene terephtalate (PET) tubes. The ISI difference between the two citrate concentrations was 5.4% for one system but not greater than 3% for all other systems when blood samples were collected with either siliconized glass or PET tubes. Most of the ISI differences between the two citrate concentrations were not significant at the 5% level. It is concluded that the ISI differences between 0.105 M and 0.109 M citrate are not of practical importance. In contrast, ISI differences between siliconised glass and PET tubes, using either 0.105 or 0.109 M citrate, were significant (p <0.05) for most thromboplastin systems and amounted to 7%. ISI interchange between these glass and PET tubes could induce INR differences amounting to 14%, which could affect clinical dosage of oral anticoagulants. PMID- 11057868 TI - Effect of factor Xa inhibitors on the platelet-derived microparticles procoagulant activity in vitro and in vivo in rats. AB - The aim of this study was to investigate the effect of factor Xa inhibitors on the prothrombinase activity of platelet-derived microparticles in vitro and in vivo. The factor Xa inhibitors studied were DX9065A (a direct factor Xa inhibitor) and Sanorg34006 (an antithrombin (AT)-mediated factor Xa inhibitor). Microparticles formed from the platelet surface following activation were isolated by size exclusion gel chromatography. After purification, their presence was detected by their procoagulant activity and by flow cytometry. Our results show that factor Xa and/or factor Va were present at the surface of the platelet derived microparticles. Prothrombinase formed on the microparticles was inhibited by factor Xa inhibitors at IC50 values of 0.45+/-0.05 and 0.045+/-0.005 microM for DX9065A and AT-Sanorg34006 respectively. In an experiment aimed at determining the kinetics of microparticles formation we demonstrated that thrombin traces were sufficient to induce the formation of a significant quantity of microparticles. Both factor Xa inhibitors delayed the formation of microparticles by delaying thrombin generation. The thrombogenic effect of the microparticles were studied in vivo in a modified arterio-venous shunt model in the rat. In this model, the increase in the thrombus weigh due to microparticles or phospholipids did not differ significantly (33% and 23% respectively). In these conditions, prothrombinase activity seemed to play a lesser role in the thrombogenic effect than phospholipids. Nevertheless, factor Xa inhibitors were efficient and inhibited thrombus formation in a dose-dependent manner. These results demonstrate that platelet-derived microparticles display a potent prothrombotic effect in vivo and show that factor Xa inhibitors are potent antithrombotic compounds when thrombosis was induced by microparticles. PMID- 11057869 TI - Histidine-rich glycoprotein (HRG) Tokushima 2: novel HRG deficiency, molecular and cellular characterization. AB - The proband, a 76-year-old woman, suffered from dural arteriovenous fistula. Her plasma histidine-rich glycoprotein (HRG) level was 50% of the normal level. A low level of plasma HRG was also found in her third daughter. A single nucleotide substitution of T to C was found at nucleotide position 11,438 in exon 6 of the HRG gene from the proband, converting Cys223 to Arg in the second cystatin-like domain. The same mutation was also identified in her third daughter, but not in the other four family members having normal HRG levels or in 50 unrelated healthy Japanese individuals. Expression studies in BHK cells showed that substantial intracellular degradation of the mutant occurred and only about 40% of the recombinant HRG mutant was secreted. These results indicate that congenital HRG deficiency caused by a substitution of Cys223 to Arg is hereditary in this family. PMID- 11057870 TI - Cerivastatin, an inhibitor of HMG-CoA reductase, inhibits urokinase/urokinase receptor expression and MMP-9 secretion by peripheral blood monocytes--a possible protective mechanism against atherothrombosis. AB - It is now recognised that acute myocardial infarction results from the rupture of atherosclerotic plaques. Lymphocytes and macrophages, which infiltrate rupture sites, contribute to plaque degradation by expressing urokinase (u-PA) bound to cell membrane by urokinase receptor (u-PAR) and by secreting metalloproteinase MMP-9. We have previously demonstrated that the uptake of oxidised LDL (ox-LDL) by monocytes induces an increase of u-PA and u-PAR expression. The present study shows that the expression of u-PA and u-PAR induced by ox-LDL on monocyte surface is suppressed by cerivastatin (a synthetic inhibitor of HMG-CoA reductase, Bayer) from 2 nM. This leads to reduced plasmin generation and monocyte adhesion to vitronectin. Furthermore, higher concentrations of cerivastatin (50-100 nM) reduce the expression of u-PA and u-PAR on unstimulated monocytes. It also inhibits MMP-9 secretion but has no effect on TIMP-1 secretion, suggesting that the decrease in MMP-9 has a real protective effect on plaque stabilisation. The inhibitory effect of cerivastatin on u-PA expression and MMP-9 secretion can be explained by the inhibition of NF-kappa B translocation into the nucleus, as shown by immunofluorescence. As farnesyl-pyrophosphate reverses the effect of cerivastatin, it is postulated that these effects could also be due to the inhibition of Ras prenylation. This was confirmed by confocal microscopy, which shows the Ras delocalisation from the monocyte membrane. The cerivastatin-induced effects on monocyte functions could explain, at least in part, the protective effect of this drug against atherothrombotic events. PMID- 11057871 TI - Platelet and platelet-derived microparticle surface factor V/Va binding in whole blood: differences between neonates and adults. AB - Platelet-derived microparticles (PDMP) appear to play a major role in the generation of procoagulant activity. In this study, we describe a novel flow cytometric method that allows direct evaluation of the procoagulant activity of PDMP and platelets in the physiological milieu of whole blood. The percent PDMP generated in response to calcium ionophore A23187 and calcium was increased in preterm neonates (67.5+/-3.4%, mean +/- S.E.M., n = 8, p <0.05) and term neonates (67.2+/-2.7%, n = 7, p<0.05) compared with adults (49.5+/-3.4%, n = 13). However, in preterm neonates A23187/calcium-induced binding of factor V/Va to PDMP and platelets (22.8+/-5.6 fluorescence units) was markedly reduced (p <0.05) compared to term neonates (58.2+/-7.2) and adults (50.6+/-6.3). In preterm blood, A23187/calcium-induced binding of factor V/Va to PDMP and platelets returned to adult levels when: a) adult plasma, rather than autologous preterm neonatal plasma, was added; or b) factor V, but not factor VIII, was added to autologous preterm neonatal plasma. In summary: 1) We have developed a flow cytometric method for the direct detection of procoagulant PDMP and platelets in whole blood. 2) Compared to adults and term neonates, PDMP and platelets of preterm neonates bound markedly less factor V/Va (reflecting reduced procoagulant activity), because of a relative lack of factor V in preterm neonates. 3) This procoagulant defect in PDMP and platelets may contribute to the propensity of preterm neonates, but not term neonates, to intraventricular hemorrhage. 4) The percent PDMP does not necessarily reflect the degree of procoagulant activity of PDMP or platelets. PMID- 11057872 TI - Regulation of platelet aggregation and adenosine triphosphate release in vitro by 17beta-estradiol and medroxyprogesterone acetate in postmenopausal women. AB - Clinical studies have suggested that hormone replacement therapy (HRT) may reduce the risk of coronary heart disease in postmenopausal women. Although progestins are commonly added to HRT preparations for uteroprotection, the perceived beneficial cardiovascular effects of HRT are thought to be mediated predominantly by the estrogen component. Platelets play a critical role in the pathogenesis of atherosclerosis and cardiovascular disease and, hence, it is possible that the cardiovascular effects of estrogens are mediated, at least in part, through inhibition of illicit platelet activation. The aim of this study was to examine the effects of sex steroids on adenosine diphosphate (ADP)-induced platelet aggregation and adenosine triphosphate (ATP) release in vitro in postmenopausal women. In addition, the effects of antiestrogens 14-hydroxy tamoxifen (4-OHT) and ICI 182780] and antiprogestins (RU 486 and ZK 98299) were also investigated. Preincubation of platelet-rich plasma (PRP) with antiestrogens or antiprogestins did not alter subsequent platelet aggregation or ATP release in response to ADP. However, preincubation with 17beta-estradiol (E2) significantly inhibited ADP mediated platelet aggregation by a mean (+/-SEM) of 37%+/-6% (p = 0.02) and ATP release by 82%+/-6% (p = 0.03), an effect that was reversed by the addition of ICI 182780 or 4-OHT but not RU 486 and ZK 98299. Although the progestin medroxyprogesterone acetate (MPA) also significantly inhibited platelet aggregation (by 28%+/-5%, p = 0.02) and ATP release (by 63%+/-9%, p = 0.02), this inhibition was not reversed by the addition of antiprogestins or antiestrogens. These data show that sex steroids can modulate platelet function in vitro. Furthermore, as platelets are devoid of nuclear components, these findings indicate that estrogens may regulate platelet function through binding to a non nuclear receptor with ligand-binding properties similar or identical to the wild type receptor. By contrast, MPA appears to exert its effect through a mechanism that does not involve binding to the "classical" progesterone receptor. PMID- 11057873 TI - Role of alpha5beta1 and alphavbeta3 integrins on smooth muscle cell spreading and migration in fibrin gels. AB - Fibrin is found at sites of vascular injury and is one of the major matrix ligands for beta3 integrins. Blocking the beta3 integrin on smooth muscle cell is hypothesized as a potential target to prevent restenosis because it could inhibit cell attachment and migration into fibrin provisional matrix. Human aortic smooth muscle cells (HNB18E6E7) spread stably in plasma gels within 24 h. Cell spreading was dramatically blocked by simultaneous use of alpha5beta1 and alphavbeta3 integrin antibodies (P <0.0001), however, blocking of either integrin alone failed to inhibit spreading. GPenGRGDSPCA, which has been considered a specific alphavbeta3 antagonist, inhibited spreading at 500 microM, suggesting that the peptide blocked both alpha5beta1 and alphavbeta3. Similarly, invasive migration into fibrin gels was blocked by simultaneous use of both alpha5beta1 and alphavbeta3 antibodies, however, blocking of either integrin alone failed to effect cell migration. Another migration assay using transwell indicated similar results. In conclusion, both alpha5beta1 and alphavbeta3 integrins are responsible for smooth muscle cell spreading and migration into fibrin gels. These data suggest that blocking beta3 integrin alone would not affect smooth muscle cell interaction with fibrin. PMID- 11057874 TI - In vivo stimulation of vascular plasminogen activator inhibitor-1 production by very low-density lipoprotein involves transcription factor binding to a VLDL responsive element. AB - High plasma levels of plasminogen activator inhibitor-1 (PAI-1) are associated with an increased risk of cardiovascular disease. There is also a close relation between high plasma levels of PAI-1 and hypertriglyceridemia. Cell culture studies have shown that very low density lipoprotein (VLDL) increases the production and secretion of PAI-1 in endothelial cells and hepatocytes, suggesting a possible mechanism for this association. To determine whether VLDL stimulates PAI-1 production in vascular cells also in vivo, Sprague-Dawley rats were injected intravenously with 6 mg/kg of VLDL (derived from human subjects with type IV hyperlipidemia). Previous studies have demonstrated that this results in an accumulation of human VLDL in the aorta and other arteries followed by increased nuclear factor-kappa B (NF-kappaB) activation. Endothelial, but not smooth muscle cells, showed a basal PAI-1 mRNA and protein expression as assessed by in situ hybridization and immunohistochemistry, respectively. Six to twenty four hours after the VLDL injection, lipoprotein particle accumulation was seen in the aortic wall, which was accompanied by increasing PAI-1 mRNA and protein expression in endothelial and smooth muscle cells. Within the rat PAI-1 promoter we identified a sequence located at -589 to -571 with 74% homology with the recently described VLDL responsive element in the human PAI-1 promoter and located adjacent to a 4-guanosine motif presumably corresponding to the human 4G/5G polymorphism. Transient transfection studies showed that VLDL exerts its stimulatory effects on rat PAI-1 gene expression in vascular cells by interaction with promoter sequences located within bp -656 and -505. Electrophoretic mobility shift assays showed that VLDL increases the binding of as yet incompletely characterized factors to this response element. Taken together these observations support a direct influence of VLDL on vascular PAI-1 gene expression ill vivo. This stimulation is exerted on the level of PAI-1 gene transcription, and involves transcription factor binding to a VLDL responsive element adjacent to a 4G motif within the PAI-1 promoter. PMID- 11057875 TI - Verotoxin-1 induces tissue factor expression in human umbilical vein endothelial cells through activation of NF-kappaB/Rel and AP-1. AB - This study examined the effect of verotoxin-1 (VT-1), which is released from Escherichia coli O157:H7, on endothelial expression of tissue factor (TF), a cofactor required to initiate blood coagulation. In order to elucidate the molecular basis for development of hemolytic uremic syndrome (HUS) in patients infected with E. coli O157:H7, human umbilical vein endothelial cells (HUVECs) were exposed to purified VT-1. VT-1 increased both TF activity and TF mRNA in HUVECs without loss of cell viability in a time- and dose-dependent manner from 0.1 to 10 ng/ml VT-1. Nuclear proteins extracted from VT-1-stimulated HUVECs bound to the consensus NF-kappaB/Rel and AP-1 binding oligonucleotides in a dose dependent manner within 2 h after the stimulation in electrophoretic mobility shift assays (EMSA). Nuclear proteins from VT-1-stimulated HUVECs formed two complexes with the NF-kappaB/Rel binding motif in the human TF promoter (TF kappaB motif). The supershift assays, using antibodies for human p65, p50 or c Rel, indicated that the lower complex was composed of p65/p50 and the higher complex was a p65 homo- or hetero-dimer with the Rel family, except c-Rel. The human TF promoter contains two AP-1 binding sites, the proximal and distal AP-1 binding sites. The supershift assays indicated that AP-1 containing mainly c-Jun and JunD, positively bound to the proximal AP-1 motif of TF (TF-AP-1). The distal TF-AP-1 motif did not show positive binding with nuclear proteins from VT-1 stimulated HUVECs. Pretreatment of HUVECs with curcumin, an inhibitor of NF kappaB/Rel activation, synthesis of c-Jun mRNA and binding of activated AP- I with AP-binding oligonucleotide, prevented the VT-1 induced increase in TF mRNA and activity in VT-1-stimulated HUVECs. Curcumin also inhibited NF-kappaB and AP 1 binding to TF-kappaB and proximal TF-AP-1 oligonucleotides, respectively, in a dose-dependent manner. The present work suggests that both the NF-kappaB/Rel and AP-1 activated in endothelial cells by stimulation with VT-1 binds to the TF kappaB and proximal AP-1 binding sites, respectively, of the TF promoter. PMID- 11057876 TI - The -5061/D polymorphism of the thrombin receptor (PAR-1) gene is not related to myocardial infarction in the ECTIM study. The Etude Cas-Temoins de l'Infarctus du Myocarde. MONICA Members Group. PMID- 11057877 TI - Prevalence of factor V Leiden mutation in Yugoslav thrombophilic patients and its relationship to the laboratory diagnosis of APC resistance. PMID- 11057878 TI - The C536T transition in the tissue factor pathway inhibitor (TFPI) gene does not contribute to risk of venous thrombosis among carriers of factor V Leiden. PMID- 11057879 TI - Risk of myocardial infarction in carriers of mutations in the hemochromatosis associated gene. PMID- 11057880 TI - von Willebrand factor ristocetin cofactor activity correlates with platelet function in a high shear stress system. PMID- 11057881 TI - Grossly abnormal proteolysis of von Willebrand factor (VWF) in a patient heterozygous for a gene deletion and mutation in the dimerization area of VWF. PMID- 11057882 TI - C-reactive protein, idiopathic venous thromboembolism and hormone replacement therapy. PMID- 11057883 TI - Soluble selectins in sepsis: microparticle-associated, but only to a minor degree. PMID- 11057884 TI - Reactivation of chronic hepatitis C virus infection by immunoadsorption in factor VIII inhibitor haemophilia. PMID- 11057885 TI - Effects of dyslipidemia on t-PA release in rats. PMID- 11057886 TI - Adult transient lupus anticoagulant patients maintain an acquired activated protein C resistance profile. PMID- 11057887 TI - Is there a role for genetic polymorphism of C677T methylenetetrahydrofolate reductase (MTHFR) in Buerger's disease? PMID- 11057888 TI - Hyperhomocyst(e)inemia, MTHFR 677C-->T polymorphism and folate status in acute coronary disease. PMID- 11057889 TI - Prevention of deep vein thrombosis after hip replacement--reply. PMID- 11057890 TI - Translocation and reversible localization of signaling proteins: a dynamic future for signal transduction. PMID- 11057891 TI - Cellular signaling: pivoting around PDK-1. PMID- 11057892 TI - Signaling reaches to new dimensions in Drosophila imaginal discs. AB - Finding that peripodial cells in wing and eye imaginal discs are essential for the growth and patterning of the separate layer of disc cells now opens the study of interacting cell layers to the powerful developmental genetic techniques with which the Drosophila system is blessed. We can anticipate that future work will identify how such interactions contribute to patterning and how the mechanisms and processes that are involved are conserved in vertebrates. We can also look forward to contributions that this work will make to understand-ing the role of interconnecting cell extensions in such signaling processes. In this minireview, we have noted numerous types of signaling cells in which cellular extensions have been observed. At present, neither the functional nor structural relationship of these related structures is known. It is certainly tempting to suggest that these structures are conduits for signals or that they function as sensors. There is, as yet, no direct experimental evidence for such roles. PMID- 11057893 TI - Signaling networks: the origins of cellular multitasking. PMID- 11057894 TI - Conservation and innovation in plant signaling pathways. PMID- 11057895 TI - Cell signaling by receptor tyrosine kinases. PMID- 11057896 TI - Ras and Rho GTPases: a family reunion. PMID- 11057897 TI - Signal transduction by the JNK group of MAP kinases. PMID- 11057898 TI - TOR, a central controller of cell growth. AB - Cell growth (increase in cell mass) and cell proliferation (increase in cell number) are distinct yet coupled processes that go hand-in-hand to give rise to an organ, organism, or tumor. Cyclin-dependent kinase(s) is the central regulator of cell proliferation. Is there an equivalent regulator for cell growth? Recent findings reveal that the target of rapamycin TOR controls an unusually abundant and diverse set of readouts all of which are important for cell growth, suggesting that this conserved kinase is such a central regulator. PMID- 11057899 TI - Signaling to chromatin through histone modifications. PMID- 11057900 TI - Insights into programmed cell death through structural biology. PMID- 11057901 TI - Signaling takes shape in the immune system. PMID- 11057902 TI - TGFbeta signaling in growth control, cancer, and heritable disorders. PMID- 11057903 TI - Linking colorectal cancer to Wnt signaling. PMID- 11057904 TI - Opposing effects of Ras on p53: transcriptional activation of mdm2 and induction of p19ARF. AB - Mdm2 acts as a major regulator of the tumor suppressor p53 by targeting its destruction. Here, we show that the mdm2 gene is also regulated by the Ras-driven Raf/MEK/MAP kinase pathway, in a p53-independent manner. Mdm2 induced by activated Raf degrades p53 in the absence of the Mdm2 inhibitor p19ARF. This regulatory pathway accounts for the observation that cells transformed by oncogenic Ras are more resistant to p53-dependent apoptosis following exposure to DNA damage. Activation of the Ras-induced Raf/MEK/MAP kinase may therefore play a key role in suppressing p53 during tumor development and treatment. In primary cells, Raf also activates the Mdm2 inhibitor p19ARF. Levels of p53 are therefore determined by opposing effects of Raf-induced p19ARF and Mdm2. PMID- 11057905 TI - Novel signaling from the peripodial membrane is essential for eye disc patterning in Drosophila. AB - The Drosophila eye disc is a sac of single layer epithelium with two opposing sides, the peripodial membrane (PM) and the disc proper (DP). Retinal morphogenesis is organized by Notch signaling at the dorsoventral (DV) boundary in the DP. Functions of the PM in coordinating growth and patterning of the DP are unknown. We show that the secreted proteins, Hedgehog, Wingless, and Decapentaplegic, are expressed in the PM, yet they control DP expression of Notch ligands, Delta and Serrate. Peripodial clones expressing Hedgehog induce Serrate in the DP while loss of peripodial Hedgehog disrupts disc growth. Furthermore, PM cells extend cellular processes to the DP. Therefore, peripodial signaling is critical for eye pattern formation and may be mediated by peripodial processes. PMID- 11057906 TI - Peripodial cells regulate proliferation and patterning of Drosophila imaginal discs. AB - Cells employ a diverse array of signaling mechanisms to establish spatial patterns during development. Nowhere is this better understood than in Drosophila, where the limbs and eyes arise from discrete epithelial sacs called imaginal discs. Molecular-genetic analyses of pattern formation have generally treated discs as single epithelial sheets. Anatomically, however, discs comprise a columnar cell monolayer covered by a squamous epithelium known as the peripodial membrane. Here we demonstrate that during development, peripodial cells signal to disc columnar cells via microtubule-based apical extensions. Ablation and targeted gene misexpression experiments demonstrate that peripodial cell signaling contributes to growth control and pattern formation in the eye and wing primordia. These findings challenge the traditional view of discs as monolayers and provide foundational evidence for peripodial cell function in Drosophila appendage development. PMID- 11057907 TI - Activation of the IkappaB kinase complex by TRAF6 requires a dimeric ubiquitin conjugating enzyme complex and a unique polyubiquitin chain. AB - TRAF6 is a signal transducer in the NF-kappaB pathway that activates IkappaB kinase (IKK) in response to proinflammatory cytokines. We have purified a heterodimeric protein complex that links TRAF6 to IKK activation. Peptide mass fingerprinting analysis reveals that this complex is composed of the ubiquitin conjugating enzyme Ubc13 and the Ubc-like protein Uev1A. We find that TRAF6, a RING domain protein, functions together with Ubc13/Uev1A to catalyze the synthesis of unique polyubiquitin chains linked through lysine-63 (K63) of ubiquitin. Blockade of this polyubiquitin chain synthesis, but not inhibition of the proteasome, prevents the activation of IKK by TRAF6. These results unveil a new regulatory function for ubiquitin, in which IKK is activated through the assembly of K63-linked polyubiquitin chains. PMID- 11057908 TI - The Na+/H+ antiporter of alkaliphilic Bacillus sp. AB - The Na+/H+ antiporter, which appears to predominantly contribute to the alkaliphily of Bacillus halodurans C-125, was studied in an alkali-sensitive mutant of this strain and a transformant with restored alkaliphily. The alkali sensitive mutant, strain 38154, which has lost the ability to grow above pH 9.5, was found to lack electrogenic Na+/H+ antiport activity driven by deltapsi (membrane potential, interior negative), and it showed defective regulation of intracellular pH under alkaline conditions. On the other hand, a transformant carrying a 2.0-kb DNA fragment from the parental genome that complemented this defect was able to maintain an intracellular pH lower than that of the external milieu, and it was found to have recovered the Na+/H+ antiport activity driven by deltapsi. Sequence analyses found that a 5.1-kb DNA region contained four open reading frames (ORF-1 to ORF-4). Direct sequencing of the corresponding region in mutant 38154 revealed a G-to-A substitution, which resulted in an amino acid substitution from Gly-393 to Arg in the putative ORF-1 product. It has been recently found that a region homologous to the DNA fragment responsible for the alkaliphily of strain C-125 exists in the genomes of Bacillus subtilis, Sinorhizobium (Rhizobium) meliloti, and Staphylococcus aureus. These homologues are present as a cluster of seven ORFs in each case. The shaA gene product of B. subtilis shows significant similarity to the ORF-1 product of strain C-125. Disruption of the shaA gene resulted in a decrease in Na+/H+ antiport activity, and growth of the shaA-disrupted strain was impaired when the external Na+ concentration was increased. We conclude that the shaA gene encodes a Na+/H+ antiporter, which plays an important role in extrusion of cytotoxic Na+. PMID- 11057909 TI - Comparisons of the polar lipid and pigment profiles of two solar salterns located in Newark, California, USA, and Eilat, Israel. AB - The whole community pigments and lipids have been examined during a 5-year period in two commercial solar salterns located in the United States and in Israel. There were significant differences in the complexity of the lipid and pigment patterns within the California saltern system, and these differences were not consistent over the sampling period despite examination of ponds with the same salinity. The solar saltern system in Eilat, Israel, showed greater consistency during this sampling period and compared directly with previous studies. The complexity of the saltern in Newark, California, could be explained on the basis of the prevailing weather conditions (cooler and more rainfall) and the nutrient enriched source water. The Eilat saltern, however, has an oligotrophic water source and has a considerably warmer and drier climate. This difference resulted in more diverse and more complex pigment and lipid patterns and presumably microbial populations in the Newark, California, plant than in the saltern in Eilat, Israel. PMID- 11057910 TI - Genetic diversity analysis of Rhodothermus reflects geographical origin of the isolates. AB - The genetic diversity and relationships of 81 Rhodothermus isolates from different geothermal environments in Iceland were examined by analysis of electrophoretically demonstrable allelic variation of 13 genes encoding enzymes. All the enzymes were polymorphic. A total of 71 distinctive multilocus genotypes (electrophoretic types, ETs) were identified. The mean genetic diversity per locus (H1) was 0.586. The relatively high genetic variance observed within Rhodothermus isolates from different locations is most likely the result of genetic changes occurring independently in the locations studied. A high Gst value (0.284) indicates that a considerable part of the variance observed is due to differences between locations. Cluster analysis revealed two major groups of ET clusters diverging at a genetic distance of 0.75, reflecting strongly the geographic origin of isolates. Estimation of the association index (I(A)) indicates that Rhodothermus marinus is a clonal species in which recombination events occur rarely. Partial or whole sequencing of the 16S rRNA genes of Rhodothermus isolates grouping at genetic distance of 0.40 confirmed that all the isolates belonged to the species Rhodothermus marinus. The results of this study confirm that, despite phylogenetic and phenotypic similarity, genetic diversity within Rhodothermus marinus is quite high. PMID- 11057911 TI - Straight-chain fatty alcohols in the hyperthermophilic archaeon Pyrococcus furiosus. AB - Two straight-chain fatty alcohols (n-hexadecanol and n-octadecanol) were found in the neutral lipid fraction extracted from Pyrococcus furiosus cells. They were identified by thin-layer and gas-liquid chromatography, mass and infrared spectra, and chemical modification. The fatty alcohols accounted for 54% of the neutral lipid of the cell. PMID- 11057912 TI - The intracellular pH of the thermophilic bacterium Thermoanaerobacter wiegelii during growth and production of fermentation acids. AB - The thermophilic glycolytic anaerobe Thermoanaerobacter wiegelii grows over the pH range 5.1-7.7, and no growth is observed below pH 5.0 or above 7.7. When T. wiegelii was grown in pH-uncontrolled batch culture, glucose was fermented to ethanol, acetate, and lactate. Small amounts of lactic acid were produced once the external pH reached 6.0, and a fructose-1.6-diphosphate (FDP) activated lactate dehydrogenase (LDH) was detected in cell-free crude extracts. Maximal activation of LDH by FDP was observed at pH 6.2. As the pH of the medium declined from 6.7 to 5.1 due to the production of acetate and lactate, the total protonmotive force (deltap) remained between - 110 and - 130mV, and the membrane potential (dekltapsi) decreased from -104 to -65mV. This decrease in deltapsi was paralleled by an increase in the chemical gradient of protons (ZdeltapH) from -31 to -62mV at pH 5.1. Based on these results, T. wiegelii maintained a small deltapH (0.3-0.9 units, inside alkaline) as the medium pH declined and interconverted deltapsi to ZdeltapH to maintain the total deltap relatively constant. Intracellular potassium decreased from 150 mM at pH 6.70 to 50 mM at pH 5.1, and this represented a 33-mV decline in the transmembrane chemical potential of potassium. The ability to synthesize ATP remained constant as the external pH declined, and therefore metabolic energy per se was not a critical aspect of pH sensitivity. PMID- 11057913 TI - Identification of facultatively alkaliphilic Bacillus sp. strain YN-2000 and its fatty acid composition and cell-surface aspects depending on culture pH. AB - Facultatively alkaliphilic Bacillus sp. strain YN-2000 was isolated from an indigo ball. Although the strain has been extensively investigated as a representative strain of alkaliphilic bacillus, its taxonomic position is not yet known. Morphological, biochemical, and physiological characteristics and chemotaxonomic properties indicated that the strain was closely related to Bacillus cohnii; this was confirmed by the high homology of the 16S rRNA sequence and the construction of a phylogenetic tree on the basis of the 16S rRNA sequence and DNA-DNA relatedness data. Strain YN-2000 contained a larger amount of unsaturated fatty acids compared with Bacillus subtilis and the obligate alkaliphile, Bacillus alcalophilus, regardless of its culture pH. When the cells were grown at pH 10, the unsaturated fatty acid content and anteiso-/iso-branched fatty acid ratio became lower than those at pH 7. This result suggests that membrane fluidity decreases when the cells are grown at pH 10 compared to those of pH 7. In the cells of strain YN-2000 grown at pH 10, the cell-surface aspect was rougher, the cell shape was longer, and the cell-surface layer was thicker compared with those of the cells grown at pH 7. The cell-surface structural change might be related to adaptation to an alkaline environment. PMID- 11057914 TI - Sucrose transport by the alkaliphilic, thermophilic Bacillus sp. strain TA2.A1 is dependent on a sodium gradient. AB - An alkaliphilic Bacillus designated strain TA2.A1, isolated from a thermal spring in Te Aroha, New Zealand, grew optimally at pH 9.2 and 70 degrees C. Sodium chloride (>5mM) was an obligate requirement for the growth of strain TA2.A1 on sucrose, and growth on sucrose was inhibited by monensin, an ionophore that collapses the sodium gradient (ApNa+) across the cell membrane. Sucrose transport by strain TA2.A1 was sodium dependent and was inhibited by monensin. The Kt for sucrose transport was 33 microM and the Eadie-Hofstee plot was linear, suggesting one high-affinity uptake system for sucrose. The affinity for sodium was low (0.5 mM), and the Hill plot had a slope of 1.6, suggesting that sodium binding was noncooperative and that the sucrose transporter had more than one binding site for sodium. Based on these results, Bacillus strain TA2.A1 uses a sodium gradient for sucrose uptake, in addition to the sodium-dependent glutamate uptake system reported previously. PMID- 11057915 TI - Cloning, expression, and characterization of pyrrolidone carboxyl peptidase from the archaeon Thermococcus litoralis. AB - The gene encoding pyrrolidone carboxyl peptidase (Pcp) has been cloned from the hyperthermophilic archaeon Thermococcus litoralis. The recombinant enzyme has been expressed in Escherichia coli, purified, and characterized. The T. litoralis Pcp demonstrates strong sequence homology to previously characterized bacterial Pcps. Some investigations have been carried out on enzyme substrate specificity and stability. PMID- 11057916 TI - A microbiological survey of Montserrat Island hydrothermal biotopes. AB - In March 1996, a survey of hydrothermal sites on the island of Montserrat was carried out. Six sites (Galway's Soufriere. Gages Upper and Lower Soufrieres, Hot Water Pond, Hot River, and Tar River Soufriere) were mapped and sampled for chemical, ATP, and microbial analyses. The hydrothermal Soufriere sites on the slopes of the active Chances Peak volcano exhibited temperatures up to almost 100 degrees C and were generally either mildly acidic at pH 5-7 or strongly acidic at pH 1.5-3, but with some hot streams and pools of low redox potential at pH 7-8. Hot Water Pond sites, comprising a series of heated pools near the western shoreline of the island. were neutral and saline, consistent with subsurface heating of entrained seawater. Biological activity shown by ATP analyses was greatest in near-neutral pH samples and generally decreased as acidity increased. A variety of heterotrophic and chemolithotrophic thermophilic organisms were isolated or observed in enrichment cultures. Most of the bacteria that were obtained in pure culture were familiar acidophiles and neutrophiles, but novel, iron-oxidizing species of Sulfobacillus were revealed. These species included the first mesophilic iron-oxidizing Sulfobacillus strains to be isolated and a strain with a higher maximum growth temperature (65 degrees C) than the previously described moderately thermophilic Sulfobacillus species. PMID- 11057917 TI - Microbiology of acidic, geothermal springs of Montserrat: environmental rDNA analysis. AB - DNA was extracted from water and sediment samples taken from acidic, geothermal pools on the Caribbean island of Montserrat. 16S rRNA genes were amplified by PCR, cloned, sequenced, and examined to indicate some of the organisms that might be significant components of the in situ microbiota. A clone bank representing the lowest temperature pool that was sampled (33 degrees C) was dominated by genes corresponding to two types of acidophiles: Acidiphilium-like mesophilic heterotrophs and thermotolerant Acidithiobacillus caldus. Three clone types with origins in low- and moderate- (48 degrees C) temperature pools corresponded to bacteria that could be involved in metabolism of sulfur compounds: the aerobic A. caldus and putative anaerobic, moderately thermophilic, sulfur-reducing bacteria (from an undescribed genus and from the Desulfurella group). A higher-temperature sample indicated the presence of a Ferroplasma-like organism, distinct from the other strains of these recently recognized acidophilic, iron-oxidizing members of the Euryarchaeota. Acidophilic Archaea from undescribed genera related to Sulfolobus and Acidianus were predicted to dominate the indigenous acidophilic archaeal population at the highest temperatures. PMID- 11057918 TI - Antidepressant drugs: a potential new drug cause of upper gastrointestinal bleeding. PMID- 11057919 TI - Helicobacter pylori cag pathogenicity island is associated with enhanced interleukin-8 expression in human gastric mucosa. AB - BACKGROUND: In vitro studies showed that Helicobacter pylori strains carrying the cag pathogenicity island are able to induce epithelial secretion of Interleukin 8. AIMS: To evaluate the assessment of cag pathogenicity island and the expression of Interleukin-8 in the gastric mucosa of Helicobacter pylori-infected patients and correlate these data with the activity of gastritis and Helicobacter pylori density. METHODS: cag status was determined by polymerase chain reaction directly on gastric biopsies from 13 Helicobacter pylori+ patients with non-ulcer dyspepsia and 13 Helicobacter pylori+ with duodenal ulcer. Interleukin-8 gene transcription and protein expression were analysed by in situ hybridization and immunofluorescence, respectively. Gastritis activity and Helicobacter pylori density were also investigated. RESULTS: cag was present in 20/26 of Helicobacter pylori+ patients: in 7/13 non-ulcer dyspepsia (53.8%] and in 13/13 duodenal ulcer patients (100%), (p<0.05). Interleukin-8 mRNA and protein expression in epithelial and inflammatory cells was higher in cag+ than in cag- patients (p<0.005). Gastritis activity significantly correlated with cag (p<0.05) and Interleukin-8 expression (p<0.005]. Helicobacter pylori density was enhanced in cag+ [p<0.005] and correlated with Interleukin-8 expression (p<0.0051. CONCLUSIONS: The present study demonstrates that in Helicobacter pylori-infected human gastric mucosa, cag+ infection is associated with enhanced Interleukin-8 expression, higher levels of active gastritis and bacterial density, and presence of duodenal ulcer. PMID- 11057920 TI - Diet and duodenal ulcer. AB - BACKGROUND: Despite the fact that the main cause of duodenal ulcer incidence and recurrence is the Helicobacter pylori bacterium, more than 80% of Helicobacter pylori-infected people never develop an ulcer. Diet may be one of the most important environmental factors contributing to duodenal ulcer. AIMS: To explore the role of diet in causation, treatment and prevention of duodenal ulcer recurrence. METHODS: All research papers published in English from 1966 to October 1999 present in Medline, involving human subjects, and having duodenal ulcer as outcome, entered the review. RESULTS AND CONCLUSIONS: Soluble fibre from fruit and vegetables seem to be protective against duodenal ulcer and refined sugars a risk factor. The role of fibre in the treatment and prevention of recurrence of duodenal ulcer is uncertain, as is that of essential fatty acids. However, none of the epidemiological studies on the relationship between diet and duodenal ulcer disease controlled for Helicobacter pylori. PMID- 11057921 TI - Clinical nutrition practice in Italian Gastroenterology Units. AB - BACKGROUND: Nutritional status affects the course, ensuing complications and prognosis of virtually all diseases. AIMS: To define the role of nutrition in Gastroenterology Units by means of two investigations that analyse: a) availability of devices for assessing nutritional status; b) nutritional treatment in clinical practice: incidence and frequency of indications for its use, together with type of treatment adopted. PATIENTS AND METHODS: Two questionnaires were sent to Italian Academic and Hospital Gastroenterology Units, all with clinical wards. RESULTS: Results refer to 27 Units, 22 of which took part in both parts of the analysis, enrolling 547 patients during the two-week study The first analysis shows that scales and the altimeter are not available everywhere, while more specific tools, such as skinfold calipers are available in 54% of the Units, and caloric intake can be assessed in 22-41%. The second analysis reveals that nutritional treatment was necessary in 50% of patients in the series examined, and that this was taken into account and prescribed in almost all cases (91%). Of the patients treated, 69% received dietetic supplementation and 31% artificial nutrition [12% enteral, 88% parenteral), although supportive parenteral nutrition is often contraindicated in conditions where good bowel function provides the conditions for enteral nutrition. CONCLUSION: Data emerging from the investigation showed that i) artificial nutrition is commonly used in gastroenterology Units in Italy although 23% of them never consider either enteral or parenteral nutrition as medical treatment of gastrointestinal disease; ii) malnutrition is a very frequent complication (mean 27%; range 4-55%0) in Gastroenterology Unit patients albeit only 42% of malnourished patients received artificial nutrition; iii) indications for enteral and parenteral nutrition are not always respected, as there is an excessive use of parenteral nutrition and an unjustified resistance to the use of enteral nutrition; iv] nutritional treatment is often administered without adequate nutritional assessment and without a complete adherence to the standards recommended for preparation of parenteral bags, supported by suitable technology; v) only two Gastroenterology Units report admitting and following patients in a home parenteral nutrition programme; vi) this investigation probably reflects the response of those Gastroenterology Units most aware of the importance of nutritional problems. Better awareness of correct practices for nutritional support should be promoted, encouraging greater use of diagnostic and monitoring techniques and a more discerning choice of the most suitable type of artificial nutrition to be administered in gastroenterology PMID- 11057922 TI - Clinical nutrition support in accredited nutritional units. PMID- 11057923 TI - Selective resistance of mucosal T-cell activation to immunosuppression in Crohn's disease. AB - BACKGROUND & AIMS: The inappropriately high state of T-cell activation found in Crohn's disease could be due to failure to respond to inhibitory signals. We tested the hypothesis that Crohn's disease mucosal T-cells are resistant to the immunosuppressive action of interleukin4. PATIENTS: Patients with Crohn's disease, ulcerative colitis, and other malignant and non-malignant conditions undergoing bowel resection. METHODS: The effect of interleukin-4 on lamina propria mononuclear cells from Crohn's disease, ulcerative colitis and control mucosa was assessed on various T-cell functions: interleukin-2-induced cytotoxicity, soluble interleukin-2 receptor and interleukin-2 production, and expression of mRNA for interleukin-2R and interferon-gamma. RESULTS: Cytotoxicity of control and ulcerative colitis cells was markedly decreased by interleukin-4, whereas Crohn's disease cells failed to be inhibited. Addition of interleukin-4 to interleukin-2-stimulated cultures decreased soluble interleukin-2R production significantly less in Crohn's disease and ulcerative colitis than control cells. In the same cultures, residual levels of interleukin-2 were significantly increased in control and ulcerative colitis, but not Crohn's disease cultures. Finally, Crohn's disease cells were significantly more resistant to interleukin-4 mediated inhibition of spontaneous and interleukin-2-induced expression of interleukin-2Ralpha and interferon-gamma mRNA compared to control cells. CONCLUSIONS: The effector function, receptor expression and cytokine production of Crohn's disease mucosal T-cells are resistant to interleukin4-mediated inhibition. Failure to respond to down-regulatory signals may contribute to persistent T-cell activation and chronicity of inflammation in Crohn's disease. PMID- 11057924 TI - IL-4 hyporesponsiveness of Crohn's disease mucosal T lymphocytes: a response of polarised Th1 cells? PMID- 11057925 TI - Crohn's disease in the elderly: clinical features and long-term outcome of 19 Greek patients. AB - AIM: To study the clinical course, prognosis, treatment and follow-up of 19 patients with Crohn's disease aged 60 years or over at the time of onset of symptoms. PATIENTS AND METHODS: A series of 12 males and 7 females aged 65 +/- 4. 9 and 66. 6 +/- 6 years at the onset of symptoms and the time of diagnosis, respectively (elderly group), were studied. Another group of patients(83 males, 53 females aged 29.8 +/- 12.4 and 32.2 +/- 12.7 at the onset of symptoms and of diagnosis, respectively) served as a control group (young group). Both groups were followed-up for a mean period of 7.2 and 9.8 years, respectively. RESULTS: The most common site of involvement in the elderly group was the distal ileum (47.4%), followed by large bowel (36.6%) and concurrent large and small bowel involvement (16%). Acute presentation was significantly more common in the older group. Fever and loss of weight were significantly less common in the older group (p<0.05). A higher rate of complications especially acute abdomen, was observed in the elderly group. There were no significant differences in the indication for surgery and type of surgical procedure applied between the two groups. However significantly fewer patients in the elderly group were operated on because of perianal abscess compared to the young group (p<0.05). Follow-up data revealed that elderly patients with Crohn's disease who had been operated upon showed no significant differences in the course of their disease compared to operated young patients. CONCLUSION: Crohn's disease in elderly persons of Greek origin follows much the same pattern as in other developed countries of the world. PMID- 11057926 TI - Familial Mediterranean fever. A review of the disease and clinical and laboratory findings in 105 patients. AB - BACKGROUND: The diagnosis of familial Mediterranean fever still remains clinical, since no specific laboratory test exists, other than a molecular genetic test which is not widely available. AIM: To evaluate the clinical findings in 105 Turkish patients; to compare these findings with those in the literature; and to make a brief review of the disease. METHODS: A total of 105 familial Mediterranean fever patients were evaluated either retrospectively (for those diagnosed before 1997), or prospectively (for those after 1997). A diagnostic criteria set was used in addition to the clinical and laboratory findings that can be seen in familial Mediterranean fever, including the newly described manifestations. Previously selected clinical and laboratory parameters were observed for three consecutive days. RESULTS: Of our patients, 88.5% were of Turkish, 3.8% of Armenian and 7. 6% of Jewish origin. Family history was positive in 87 (82.8%) patients. Involved site was peritoneum in 97 (92%), joints in 45 (42.8%) and pleura in 19 (18%). Frequency of myalgia/arthralgia was 24.7%, and skin findings were observed in 16. 1% of patients. Splenomegaly, not related to amyloidosis, was present in 21 (20%) patients. Meningeal, retinal or ovarian/testicular involvement was not observed. CONCLUSION: Identification of familial Mediterranean fever gene has led to the application of a molecular genetic test for the diagnosis of Familial Mediterranean Fever. Until genetic methods become widely available, diagnosis will remain clinical. Thus, awareness of various clinical forms and of the correct usage of diagnostic criteria in various patient populations is important. PMID- 11057927 TI - The augmenter of liver regeneration induces mitochondrial gene expression in rat liver and enhances oxidative phosphorylation capacity of liver mitochondria. AB - BACKGROUND: The mammalian augmenter of liver regeneration gene encodes a protein involved in the unique process of liver regeneration. The augmenter of liver regeneration respective protein stimulates hepatocyte proliferation in hepatectomized rats and inhibits cytotoxic activity of liver-derived Natural Killer cells from intact rats. Augmenter of liver regeneration protein shares homology with a Saccharomyces Cerevisiae protein essential for the viability, oxidative phosphorylation and cell-division cycle. AIMS: To demonstrate if augmenter of liver regeneration protein, like the homologous in the yeast, plays a role in the regulation of biogenesis of mitochondria. METHODS: Augmenter of liver regeneration protein was injected in intact rats and, in the hepatic tissue, the expression of two genes located in two different regions of the mitochondrial genome, mitochondrial ATPase 6/8, and ND1 subunit, and of a nuclear gene, mitochondrial Transcription Factor A, were considered. In addition, cytochrome content and oxidative phosphorylation capacity of liver-derived mitochondria were evaluated. RESULTS: The augmenter of liver regeneration protein administration induces an increase in the mitochondrial gene expression and enhances cytochrome content and oxidative phosphorylation capacity of liver derived mitochondria. CONCLUSIONS: The present data demonstrate a comparable role in the regulation of mitochondria biogenesis in the eukaryotic cell like the yeast protein. This phenomenon could be part of the complex mechanism through which augmenter of liver regeneration regulates hepatocyte proliferation. PMID- 11057928 TI - Azathioprine or 6-mercaptopurine for inflammatory bowel disease: do risks outweigh benefits? AB - The treatment of Crohn's disease and ulcerative colitis has evolved and has improved the quality of life of patients afflicted with these disorders. Immune modulators such as azathioprine and 6-mercaptopurine are an important class of medications used for the treatment of patients with inflammatory bowel disease. Controlled studies have demonstrated their efficacy in both induction and maintenance of remission in Crohn's disease, and similarly, for the induction and maintenance of remission in patients with ulcerative colitis. These agents have had an increasing importance in the management of steroid-resistant, steroid dependent diseases, and fistulizing Crohn's disease. The primary limitations to these agents have been their slow onset of action and their side effect profile. Despite these limitations, these agents have demonstrated efficacy and have become paramount to the management of patients with these incurable potentially disabling disorders. The precise role of azathioprine/6-mercaptopurine, their limitations and their safety are reviewed in this paper. PMID- 11057929 TI - Clinical use of manometry for the diagnosis of intestinal motor abnormalities. AB - Digestive symptoms suggestive of intestinal motor disorders, such as abdominal pain and distension, fullness, vomiting, constipation and diarrhoea, are very common and non-specific, and may be clinical manifestations of both organic and functional diseases. Both radiology and endoscopy are important in the diagnosis of structural gastrointestinal lesions that can affect motility and offer indirect signs of impaired gastrointestinal functions, but the diagnosis of gut motility disorders currently relies on the manometric assessment of contractile activities. Small bowel manometry helps to identify normal motility features and consequently to identify abnormal motor patterns. Small bowel manometry can help to differentiate mechanical obstruction from pseudo-obstruction and neurogenic from myogenic motor disorders. Manometry is an invasive technique which is not well accepted by patients and requires specific skills from investigators. Also, manometric assessment is limited to referral centres with a specific interest in the field of digestive functions, in general, and motility, in particular. Only patients who remain undiagnosed after extensive traditional work-up and fail repeated courses with medical therapy should be referred for the manometric test. Understanding the underlying pathophysiologic mechanisms of abnormal motility and developing new therapies are the goals of the current research in this fascinating field of medicine. PMID- 11057930 TI - Inhibition of the cyclooxygenase/lipoxygenase pathways to improve interferon alpha efficacy in chronic hepatitis C: don't lose the tract! PMID- 11057931 TI - Chronic atrophic gastritis and gastric cancer. PMID- 11057932 TI - Antiretroviral therapy for drug users. AB - Injection drug use represents the primary risk factor for up to 40% of patients with HIV infection. Physicians are generally reluctant to prescribe antiretroviral therapy (ART) for these patients due to possible poor adherence, and the potential for complex drug interactions to occur. Providing daily observed ART in conjunction with methadone maintenance therapy (MMT) has significantly improved accessibility of ART for many drug users. Knowledge of potential drug interactions between methadone, ART, and both legally and illegally prescribed drugs has permitted such interactions to be anticipated and either avoided or treated appropriately. Optimizing ART for drug users therefore demands a multidisciplinary approach from medical, clinical pharmacology and psychiatric services. PMID- 11057933 TI - Blood-borne virus infection: the occupational risks. AB - Healthcare workers who are in contact with patients and/or clinical material are at continuous risk of acquiring blood-borne viral infections, in particular, infection with hepatitis B virus, hepatitis C virus and the human immunodeficiency virus (HIV). Prevention of occupational exposure is dependent on education. Prevention strategies include immunization, exposure avoidance by the use of universal precautions at all times, and post-exposure advice and prophylaxis. This article will review the risks to healthcare workers associated with exposure to hepatitis B, hepatitis C, and HIV. It will discuss post-exposure management in the United Kingdom (UK) and briefly outline the risks to patients from infected healthcare workers. PMID- 11057934 TI - Safety of oral versus intravenous hydration during induction therapy with intravenous foscarnet in AIDS patients with cytomegalovirus infections. AB - We undertook a study to compare the safety of intravenous (i.v.) versus oral hydration to prevent nephrotoxicity associated with the use of foscarnet for induction therapy of cytomegalovirus (CMV) infection in HIV-infected persons. Patients, given foscarnet at a dose of 90 mg/kg every 12 h, were randomized to receive either i.v. or oral hydration. Thirty-seven patients were given i.v. hydration and 44 were given oral hydration. Median duration of therapy for both groups was 17 days. There was no difference between the 2 groups in either serious adverse events or rise of creatinine to > or = 2.0 mg/dl. However, serum creatinine, while generally remained within normal limits, increased more in patients who received oral hydration after 10 days of therapy (significant only by slope analysis, P < 0.05). Although i.v. hydration provided better protection against nephrotoxicity, oral hydration was relatively safe and convenient provided that creatinine clearance (CrCl) is monitored closely. PMID- 11057935 TI - Cervical cytology screening for sexually-active teenagers. AB - Our aim was to determine the value of cervical cytology for sexually-active teenagers attending genitourinary medicine (GUM) clinics. This is a retrospective review of 57,093 women who had cervical cytology reported at the Royal Bolton Hospital (RBH) during the period 1 April 1994 to 31 March 1996. The results were subdivided into 2 age groups; < or = 19 years and > or = 20 years. Cervical cytology abnormalities within each age group were analysed and compared with the national figures for cervical cytology screening. The results were also subdivided according to their source of referral; general practitioners (GP), GUM and gynaecology. The data from each group were analysed in the same manner as before. The results of the cervical cytology screening from the RBH for major cytological abnormalities (moderate dyskaryosis and above), for the 2 age groups, were comparable and not significantly different (2.1% for each age group, P=0.909) and these figures were within the range of the national figures for cervical cytology screening (1.1%-3.4%). Subdivision of the data by source of referral demonstrated that the percentage of major abnormalities in smears performed at GUM clinics was higher than the national figure for all age groups (41/1000 for the younger age group and 37/1000 for the older age group, 12/1000 for the national figures). In conclusion, cervical cytology screening should be offered to sexually-active teenagers attending GUM clinics. PMID- 11057936 TI - Chlamydia trachomatis PCR (Cobas Amplicor) in women: endocervical specimen transported in a specimen of urine versus endocervical and urethral specimens in 2-SP medium versus urine specimen only. AB - The sensitivity of Roche Cobas Amplicor Chlamydia trachomatis polymerase chain reaction (PCR) including the internal control (IC) programme to identify inhibition, was investigated on 3 different samples from women: (1) swab samples from the urethra and the cervix pooled in 2-SP transport medium, (2) swab sample from the cervix transported in a urine sample from the same patient, and (3) urine sample alone. Out of the 2412 patients, 193 (8.0%) were chlamydia positive and in 14 of these the results showed discrepancies between sampling methods. The sensitivity of PCR on urethra/cervix, urine/cervix and urine was 98.4% (190/193), 97.9% (189/193) and 93.3% (180/193) respectively. The higher sensitivity of PCR on urethra/cervix and urine/cervix as compared with urine alone was statistically significant. Without the IC, the sensitivity of PCR on urethra/ cervix, urine/cervix and urine would have been 95.9% (185/193), 94.8% (183/193) and 90.7% (175/193) respectively. Factors influencing the rate of inhibition were also studied. PMID- 11057937 TI - Epoetin alfa therapy for anaemia in HIV-infected patients: impact on quality of life. AB - To evaluate the effect of epoetin alfa on the quality of life (QOL) of HIV infected patients in the community setting, 221 anaemic (haemoglobin < or = 11 g/dl) HIV-positive patients from community-based treatment centres and physicians' offices were treated with epoetin alfa (100-300 units/kg subcutaneously 3 times a week) in a 4-month, open-label, non-randomized, phase IV trial. Epoetin alfa therapy significantly (P<0.01) increased and maintained haemoglobin levels (mean increase=2.5 g/dl; n=207); the improvement in haemoglobin levels was independent of changes in CD4+ cell counts. Transfusion requirements were also significantly reduced from 20% to 5% of patients (P<0.01). Mean total QOL score measured by the Functional Assessment of HIV Infection (FAHI) scale and Physical Well-Being subscale score improved significantly (P<0.05). QOL improvements associated with increases in haemoglobin were independent of changes in CD4+ counts and baseline anaemia severity. Adverse events observed during epoetin alfa therapy were consistent with HIV disease and not likely due to the drug. Epoetin alfa therapy should be considered a treatment option for HIV-infected patients with mild-to-moderate anaemia. PMID- 11057938 TI - Asymptomatic gonorrhoea and chlamydial infection in a population-based and work site based sample of men in Kilimanjaro, Tanzania. AB - The aim of this study was to screen healthy rural and urban Tanzanian men for chlamydial infection and gonorrhoea, and determine the prevalence and the predictive value of urethral symptoms, signs and pyuria. In 2 cross-sectional surveys, 796 men were interviewed regarding symptoms and examined for signs of urethritis. Gonorrhoea was detected by culture/gram-stained smears, Chlamydia trachomatis by antigen immunoassay, and pyuria by leukocyte esterase dipstick test. The prevalence of chlamydial infection, gonorrhoea and pyuria among rural men was 9.6%, 0.4%, and 12.7%, and among urban bar workers 7.4%, 8.1% and 6.3% respectively. Among all, 0.6% had urethral discharge confirmed by examination, while 2.6% reported urethral discharge and 7.4% dysuria. Among chlamydia-infected men, 59 (89%) of the 66 cases did not have urethritis symptoms or signs. Similarly, 24 (88%) of 28 men with gonorrhoea were asymptomatic. Treatment based on the urethral discharge sign, would have detected only one out of 92 cases with gonorrhoea and/or chlamydia in these populations. PMID- 11057939 TI - The use of D-dimers in the diagnosis of occult pulmonary emboli in HIV pulmonary disease--two case reports. AB - Pulmonary thromboembolism is not considered a common cause of morbidity in HIV disease. Although anti-phospholipid antibodies are often seen in HIV disease, they are not associated with an increased thrombotic risk. Computed tomographic (CT) pulmonary angiography has been described as the imaging modality of first choice, as abnormal baseline chest X-rays may reduce the diagnostic utility of ventilation perfusion (VIQ) scanning. In HIV-negative individuals D-dimer testing has been shown to be a good screening tool in suspected pulmonary embolism. We present 2 cases where the diagnosis of pulmonary embolus was established using tests for the clotting degradation products D-dimers. PMID- 11057940 TI - Using a national guideline: an audit of the management of gonorrhoea in Newcastle upon Tyne. AB - The management of cases of gonorrhoea in the Genitourinary Medicine Department (GUM) of Newcastle upon Tyne in 1999 was audited. The sensitivity of microscopy in diagnosis was 90.4% for male urethral gonorrhoea and 26.6% for female genital gonorrhoea. The sensitivity of laboratory culture was 98.5%. Screening for Chlamydia trachomatis co-infection or treating for presumed co-infection was done in 98% of cases of gonorrhoea and the rates of co-infection were 36.4% in women and 8% in men. Effective first-line therapy was given to 95.4% of cases of genital gonorrhoea. Re-attendance for tests of cure within one month of treatment was achieved in 68.2%. Discussion of partner notification was documented in 94.3% of cases of genital gonorrhoea. In 65% of cases of gonorrhoea, there was documented attendance of sexual partners. The value of a national guideline with auditable targets is discussed. PMID- 11057941 TI - The fascinating development of sexually transmitted diseases. PMID- 11057942 TI - Exacerbation of hepatitis B infection following cessation of lamivudine therapy. PMID- 11057943 TI - Focal adenitis developing after immune reconstitution with HAART. PMID- 11057944 TI - Human papillomavirus in the vaginal introitus in women infected with the human immunodeficiency virus. PMID- 11057945 TI - Clinical evidence for the adenoma-carcinoma sequence, and the management of patients with colorectal adenomas. AB - A large body of clinical evidence supports the belief that over 95% of colorectal cancers arise in benign adenomatous polyps that develop and grow very slowly over many years. Interruption of the adenoma-carcinoma sequence by resecting adenomatous polyps is a powerful method of secondary prevention of colorectal cancer. Colonoscopy is the procedure of choice for the diagnosis and resection of colorectal polyps. Patients who have had colonoscopic resection of adenomas, and in some cases their close relatives, are at increased risk for developing metachronous polyps and cancer and may benefit from follow-up colonoscopic surveillance. This surveillance should be individually tailored to the perceived risk of each case depending on the features of the adenomas removed and other patient factors such as family history. Widespread adoption of current postpolypectomy guideline recommendations is protective and conserves medical resources. PMID- 11057946 TI - Colonic adenomas: prevalence and incidence rates, growth rates, and miss rates at colonoscopy. AB - Cost-effective use of colonic imaging studies can be achieved through an understanding of the prevalence, incidence, growth, and miss rates of colon adenomas. High adenoma prevalence rates are associated with increasing age, male gender, and a family history of colorectal cancer or multiple first-degree relatives with colorectal neoplasia or neoplasia diagnosed at a young age. The incidence rate of advanced adenomas is higher in patients with multiple adenomas and is likely also associated with a family history of colorectal cancer, increased age, and large adenoma size at the index examination. Direct observational data on growth rates suggests that adenomas <1 cm in size have a fairly stable size over a 3-year interval. Although colonoscopy is the most sensitive colonic imaging study, substantial miss rates for small adenomas are inherent to the procedure. Advancements in endoscopic technology should lead to reduced miss rates, allowing expansion of intervals between examinations while reducing negative outcomes of fatal interval cancers. PMID- 11057947 TI - Genomic events in the adenoma to carcinoma sequence. AB - Owing to its high incidence and anatomical accessibility, colorectal cancer has become the most extensively studied human neoplasm with respect to molecular pathogenesis. Many of the genomic alterations that occur when a normal colonic epithelial cell gradually is transformed into a malignant cell have been well characterized. An understanding of the molecular basis of colorectal cancer will lead to better cancer control through novel and scientifically based cancer risk assessment, diagnostics, prognostics, and therapeutics. It is imperative that clinicians possess a basic understanding of the molecular pathogenesis of colorectal cancer so that they are poised to embrace new, molecularly based, preventive and treatment measures. This task is formidable given the rapidly expanding body of scientific knowledge on the genomics of colorectal cancer. PMID- 11057948 TI - Genetic syndromes and genetic tests in colorectal cancer. AB - Our understanding of the biology of colon cancer has matured to the point that it is a useful general paradigm for understanding solid tumor development. Recent advances provide insight into the genetic alterations underlying the development of colon cancer. These insights provide unique opportunities for genetic testing in predisposed, asymptomatic patients that can direct screening efforts and their clinical management. This review examines several inherited colon cancer predispositions, well described clinically for a century, that are now amenable to genetic testing. Additional discussion focuses on colon cancer predisposition traits that occur with high frequency but low penetrance characteristics. Finally, genetic tests for tumor markers that potentially have prognostic or therapeutic implications are reviewed. PMID- 11057949 TI - Interrupting the adenoma-carcinoma sequence: screening for adenomas and cancer, now and in the future. AB - Screening for colorectal cancer is only beginning to receive the attention it deserves. As screening emerges into the mainstream and utilization increases, competing technologies will battle to assume a greater percentage of the market share of testing. In this review, the standards and principles with which screening tests are evaluated are outlined. Current modalities for screening, including fecal occult blood testing (FOBT), flexible sigmoidoscopy, colonoscopy, barium enema, and the combination of FOBT with sigmoidoscopy, are discussed and critically reviewed. New techniques and technologies for screening, including virtual colonoscopy and molecular methods of screening stool, are previewed. Increased attention on screening and the competition for supremacy among the modalities undergoing evaluation make the prospects for a continued diminution in colorectal cancer mortality promising. PMID- 11057950 TI - Pathology of the adenoma-carcinoma sequence: from aberrant crypt focus to invasive carcinoma. AB - The adenoma-carcinoma sequence postulates that colorectal carcinomas arise from precursor lesions, called adenomas. All adenomas contain dysplastic epithelium that arises from mutations in either the adenomatous polyposis coli gene or DNA mismatch repair genes. The earliest lesion detected with dysplasia is the aberrant crypt focus. Over time, as this lesion acquires additional mutations, it evolves into a classic adenomatous polyp. Adenomatous polyps are classified as tubular, tubulovillous, or villous. Generally, as polyps increase in size, the degree of dysplasia worsens, the villous component increases, the number of genetic abnormalities increases, and the likelihood of harboring invasive carcinoma increases. Carcinomas associated with DNA mismatch repair mutations are more likely to be poorly differentiated and incite a host lymphocytic response. These tumors seem to have a better prognosis, stage for stage, than typical colorectal carcinomas. PMID- 11057951 TI - Flat neoplasms in the adenoma-carcinoma sequence in Japan. AB - Flat adenomas have a higher risk of association with adenocarcinoma than the protruded type, and flat cancer has a tendency to invade deeply despite its small size. Furthermore, it has been suggested that flat, small carcinomas readily invade through the muscularis mucosa into the submucosa and are associated with an increased risk of early metastasis into lymph nodes, as compared with polypoid growth pattern cancers. It has become evident that these lesions are becoming more common in Japan, as well as in some western countries. It has been generally believed that the colon is comparable to the stomach in terms of endoscopic diagnostic capability and accuracy. Therefore, we were surprised to learn that flat neoplasms, including flat or depressed adenomas and cancers, exist in the colon, as they seem to have previously been overlooked. Recent significant advances in endoscopic resolution and the development of improved diagnostic techniques and modalities may play an important role in the detection of such lesions. In the present study, the diagnosis and management of such lesions, along with their genetic background, is reviewed. Some techniques for their detection currently being used in Japan will also be reviewed using several actual cases. PMID- 11057952 TI - The pandemic of Salmonella enteritidis phage type 4 reaches Utah: a complex investigation confirms the need for continuing rigorous control measures. AB - In 1995, Salmonella Enteritidis (SE) cases in the state of Utah increased fivefold. Isolates were identified as phage type 4 (PT4). Risk factors and sources of infection were investigated in two case-control studies, a traceback of implicated foods, and environmental testing. Forty-three patients with sporadic infections and 86 controls were included in a case-control study of risk factors for infection. A follow-up case-control study of 25 case and 19 control restaurants patronized by case and control patients examined risks associated with restaurant practices. In the first case-control study, restaurant dining was associated with illness (P = 0.002). In the follow-up case-control study, case restaurants were likelier to use > 2000 eggs per week (P < 0.02), to pool eggs (P < 0.05), and to use eggs from cooperative 'A' (P < 0.009). Eggs implicated in separately investigated SE PT4 outbreaks were traced to cooperative 'A', and SE PT4 was cultured from one of the cooperative's five local farms. We conclude that SE PT4 transmitted by infected eggs from a single farm caused a fivefold increase in human infections in Utah. PMID- 11057953 TI - An outbreak of Salmonella blockley infections following smoked eel consumption in Germany. AB - In June 1998, an increased number of persons with Salmonella blockley infection were reported from one German state. Because S. blockley is extremely uncommon in Germany, a case-control study was performed in order to find the source. A total of 13 patients met the case definition. Nine of 12 cases and 2 of 21 controls with food consumption histories reported eating smoked eel (OR 28.5; 95% CI 3.9 235.3). The consumed eel came from four different local smokeries, but could be traced back to fish farms in Italy. This outbreak indicates that eel may be a vehicle for salmonella infection and that the smoking process may not eliminate bacterial contamination from raw fish. PMID- 11057954 TI - Epidemiologic application of pulsed-field gel electrophoresis to an outbreak of Campylobacter jejuni in an Austrian youth centre. AB - We report the first documented Campylobacter jejuni outbreak in an Austrian youth centre. Sixty-four children were involved of which 38 showed classical signs of campylobacter gastroenteritis. Since unpasteurized milk distributed by a local dairy was suspected to be the source of infection, stool samples were collected from 20 cows providing the milk. Five of the cows tested positive for C. jejuni. These isolates together with 37 clinical samples were compared by pulsed-field gel electrophoresis (PFGE). The PFGE patterns, using the restriction endonucleases SmaI and SalI, were identical for the human and bovine isolates. This finding confirmed that the outbreak was caused by the consumption of unpasteurized milk contaminated with C. jejuni. PMID- 11057955 TI - Clonal dissemination of Vibrio parahaemolyticus displaying similar DNA fingerprint but belonging to two different serovars (O3:K6 and O4:K68) in Thailand and India. AB - Active surveillance of Vibrio parahaemolyticus infection among hospitalized patients in Calcutta, India, showed the appearance of the O4:K68 serovar for the first time in March 1998 alongside the continued predominant incidence of the O3:K6 serovar. Strains belonging to both these serovars have been reported to possess pandemic potential. The genomes of O3:K6 and O4:K68 strains and for comparison, non-O3:K6 and non-O4:K68 strains isolated from two different countries, India and Thailand, were examined by different molecular techniques to determine their relatedness. The O3:K6 and O4:K68 strains from Calcutta and Bangkok carried the tdh gene but not the trh gene. Characterization of representative strains of these two serovars by ribotyping and by arbitrarily primed-polymerase chain reaction (AP-PCR) showed that the isolates had identical ribotype and DNA fingerprint. Pulsed-field gel electrophoresis (PFGE) performed with the same set of strains yielded nearly similar restriction fragment length polymorphism (RFLP) patterns for the O3:K6 and O4:K68 isolates from Calcutta and Thailand. Phylogenetic analysis of the NotI RFLP showed that the O3:K6 and O4:K68 strains formed a cluster with 78-91% similarity thus indicating close genetic relationship between the two different serovars isolated during the same time frame but from widely separated geographical regions. The non-O3:K6 and non O4:K68, in contrast, showed different ribotype, AP-PCR and PFGE patterns. PMID- 11057956 TI - Detection of virulence associated genes, haemolysin and protease amongst Vibrio cholerae isolated in Malaysia. AB - Eighty-four strains of Vibrio cholerae O1, O139 and non-O1/non-O139 from clinical and environmental sources were investigated for the presence of the toxin co regulated pilus gene, tcpA, the virulence cassette genes ctxA, zot, ace and cep and also for their ability to elaborate haemolysin and protease. The ctxA and zot genes were detected using DNA-DNA hybridization while the ace, cep and tcpA genes were detected using PCR. Production of haemolysin and protease was detected using mammalian erythrocytes and an agar diffusion assay respectively. Analysis of their virulence profiles showed six different groups designated Type I to Type VI and the major distinguishing factor among these profiles was in the in vitro production of haemolysin and/or protease. Clinical O1, O139 and environmental O1 strains were similar with regard to presence of the virulence cassette genes. All environmental O1 strains with the exception of one were found to possess ctxA, zot and ace giving rise to the probability that these strains may actually be of clinical origin. One strain which had only cep but none of the toxin genes may be a true environmental isolate. The virulence cassette and colonization factor genes were absent in all non-O1/non-O139 environmental strains but production of both the haemolysin and protease was present, indicating that these may be putative virulence factors. These findings suggest that with regard to its pathogenic potential, only strains of the O1 and O139 serogroup that possess the tcpA gene which encodes the phage receptor, have the potential to acquire the CTX genetic element and become choleragenic. PMID- 11057957 TI - Estimation of the under-reporting rate for the surveillance of Escherichia coli O157:H7 cases in Ontario, Canada. AB - Two models estimating the proportion of Escherichia coli O157:H7 cases not reported in the Ontario notifiable diseases surveillance system are described. The first model is a linear series of adjustments in which the total number of reported cases is corrected by successive underreporting coefficients. The structure of the second model is based on a relative difference in the proportion of E. coli O157:H7 cases which are hospitalized between the surveillance database and the underlying population. Based on this analysis, the rate of under reporting of symptomatic cases of E. coli O157:H7 infection in Ontario ranges from 78 to 88% corresponding to a ratio of 1 reported case for approximately 4-8 symptomatic cases missed by the surveillance system. This study highlights the need to increase awareness among public health workers of the potential biases that may exist in the interpretation of routine surveillance data. PMID- 11057958 TI - Investigation of human infections with verocytotoxin-producing strains of Escherichia coli (VTEC) belonging to serogroup O118 with evidence for zoonotic transmission. AB - Twenty verocytotoxigenic Escherichia coli (VTEC) O118 strains isolated between 1996 and 1998 from human patients in Germany were analysed for their serotypes, their virulence markers and their epidemiological relatedness. Three strains were typed as O118:H12, these carried only the VT2d-Ount variant gene and were not associated with diarrhoea or haemolytic uraemic syndrome (HUS). Seventeen strains were serotyped as O118:H16 or O118:non-motile (NM). These carried all the genes for VTI, eae and EHEC-haemolysin. The O118:H16/NM strains were from diarrhoea (13 cases) and HUS (2 cases). Sixteen of the patients were young infants and most infections were associated with a rural environment. Evidence for zoonotic transmission from cattle to humans was found in two cases. The epidemiological relationship between the human and bovine O118:H16/NM isolates was indicated by homogeneous plasmid patterns and by very similar XbaI restriction patterns obtained by pulsed-field gel electrophoresis. VTEC O118:H16/NM are emerging pathogens in Germany and should be classified as new enterohaemorrhagic E. coli (EHEC) types. PMID- 11057959 TI - Detection and characterization of Shiga toxin-producing Escherichia coli from seagulls. AB - Shiga toxin (Stx)-producing Escherichia coli (STEC) strains isolated from a seagull in Japan were examined. A total of 50 faecal samples was collected on a harbour bank in Hokkaido, Japan, in July 1998. Two different STEC strains, whose serotypes were O136:H16 and O153:H-, were isolated from the same individual by PCR screening; both of them were confirmed by ELISA and Vero cell cytotoxicity assay to be producing active Stx2 and Stx1, respectively. They harboured large plasmids, but did not carry the haemolysin or eaeA genes of STEC O157:H7. Based on their plasmid profiles, antibiotic resistance patterns, pulsed-field gel electrophoresis analysis (PFGE), and the stx genes sequences, the isolates were different. Phylogenic analysis of the deduced Stx amino acid sequences demonstrated that the Stx toxins of seagull-origin STEC were closely associated with those of the human-origin, but not those of other animal-origin STEC. In addition, Stx2phi-K7 phage purified from O136 STEC resembled Stx2phi-II from human-origin O157:H7, and was able to convert non-toxigenic E. coli to STEC. These results suggest that birds may be one of the important carriers in terms of the distribution of STEC. PMID- 11057960 TI - Interpretation of band differences to distinguish strains of Serratia marcescens by pulsed-field gel electrophoresis of XbaI DNA digests. AB - The number of band differences in DNA macrorestriction profiles required to distinguish unrelated strains from an index strain varies in an outbreak with the species and restriction enzyme used. In order to define this difference for epidemiological studies of Serratia marcescens, we produced DNA fingerprints from 57 isolates of the organism using the restriction enzyme XbaI and pulsed-field gel electrophoresis (PFGE). The isolates were selected on the basis of their epidemiology, serotype and phage-typing patterns to include 28 unrelated strains and 29 representatives from 2 distinct outbreaks. One of the outbreaks was prolonged. lasting for several years. Electrophoretic profiles consisting of 20 or more clearly resolved bands were obtained for all isolates. Twenty-six of the unrelated strains had unique profiles with over 10 band differences from all other strains, while 27 of the outbreak representatives could be assigned to the appropriate outbreak with confidence. The majority of the outbreak isolates had none or 2 band differences from the index profile, although 3 isolates differed by 5-7 bands. The 2 exceptions among the unrelated strains differed by 4 bands, and 3 phage typing reactions, and were isolated from London and Berlin 3 years apart, while the 2 exceptions among the outbreak collection had clearly unique profiles with over 20 band differences from each other and the outbreak profiles. Cluster analysis using Dice coefficient and UPGMA gave cut-off values of 75-78% similarity overall for related isolates, while the closest similarity for unrelated strains was 70%. The results of this study together with those of the 6 previous reports of PFGE for S. marcescens (which used either enzymes XbaI or SpeI) confirm that this technique is of value for this species and that with XbaI at least, most epidemiologically related strains will only differ by 3-4 bands. However, on occasion up to 7 band differences can be found within an apparent outbreak, which may be suggestive of genetic drift. PMID- 11057961 TI - Characterization of Klebsiella terrigena strains from humans: haemagglutinins, serum resistance, siderophore synthesis, and serotypes. AB - Klebsiella terrigena is very rarely isolated from humans; as yet, its clinical significance is uncertain. The aim of the present study was to evaluate whether this species is able to express putative virulence factors. A total of 72 faecal (n = 50) and clinical (n = 22) K. terrigena isolates was investigated and compared with faecal and clinical strains of K. pneumoniae. Mannose-sensitive haemagglutination (MSHA) was observed less often in K. terrigena (64-74%) than in K. pneumoniae strains. In contrast, the incidence of mannose-resistant haemagglutinin indicative of type 3 pili (MR/K-HA) (77-94%), serum resistance properties (10-23%), and production of enterobactin (100%) was similar in both species. None of the K. terrigena isolates were able to synthesize aerobactin; however, the frequency of aerobactin synthesis in K. pneumoniae was also only 5%. Serotyping showed capsular types K5 and K70 to be predominant. The virulence associated serotype K2 was common in both K. terrigena and K. pneumoniae isolates. Taken together, the present results suggest that K. terrigena and K. pneumoniae are indistinguishable with respect to the expression of virulence factors. PMID- 11057962 TI - Contaminated drinking water in one town manifesting as an outbreak of cryptosporidiosis in another. AB - In early 1992 we identified an outbreak of cryptosporidiosis in Oregon and sought to identify and control its source. We used a series of studies to identify risk factors for illness: (i) a case-control study among employees of a long-term-care facility (LTCF); (ii) a matched case-control study of the general community; (iii) a cohort study of wedding attendees; and (iv) a cross-sectional survey of the general community. Drinking Talent water was associated with illness in the LTCF (OR = 22.7, 95 % CI = 2.7-1009.0), and in the community (matched OR = 9.5, 95% CI 2.3-84.1). Drinking Talent water was associated with illness only among non-Talent residents who attended the wedding (P < 0.001) and in the community (RR = 6.5, 95 % CI 3.3-12.9). The outbreak was caused by contaminated municipal water from Talent in the absence of a discernible outbreak among Talent residents, suggesting persons exposed to contaminated water may develop immunity to cryptosporidiosis. PMID- 11057963 TI - A serological survey of college students for antibody to Cryptosporidium before and after the introduction of a new water filtration plant. AB - In April 1997, a large city in the northeastern United States changed their drinking water treatment practices. The city, which previously provided only chlorination for their surface water sources added filtration in addition to chlorination. To assess whether Cryptosporidium infections rates declined following filtration, we tested serological responses to 15/17-kDa and 27-kDa Cryptosporidium antigens among 107 community college students 1 month before and 225 students 5 months after filtration. Results suggest that levels of Cryptosporidium infections did not decline following water filtration. However, seasonal changes in other exposures may have confounded the findings. Swimming in a lake, stream or public pool and drinking untreated water from a lake or stream predicted a more intense response to one or both markers. Residence in the city, not drinking city tap water or drinking bottled water, gender, travel or exposure to pets, young pets, diapers or a household child in day care were not found to be predictive of more or less intense serological responses for either the 15/17 kDa and 27-kDa antigen. PMID- 11057964 TI - Widespread environmental contamination with Norwalk-like viruses (NLV) detected in a prolonged hotel outbreak of gastroenteritis. AB - A protracted outbreak of Norwalk-like virus (NLV)-associated gastroenteritis occurred in a large hotel in North-West England between January and May 1996. We investigated the pattern of environmental contamination with NLV in the hotel during and after the outbreak. In the ninth week, 144 environmental swabs taken from around the hotel were tested for NLV by nested RT-PCR. The sites were categorized according to the likelihood of direct contamination with vomit/faeces. The highest proportion of positive samples were detected in directly contaminated carpets, but amplicons were detected in sites above 1.5 m which are unlikely to have been contaminated directly. The trend in positivity of different sites paralleled the diminishing likelihood of direct contamination. A second environmental investigation of the same sites 5 months after the outbreak had finished were all negative by RT-PCR. This study demonstrates for the first time the extent of environmental contamination that may occur during a large NLV outbreak. PMID- 11057965 TI - Carriage of N. lactamica in a population at high risk of meningococcal disease. AB - Carriage of Neisseria lactamica among household contacts of meningococcal disease (MCD) cases was investigated during an epidemic in Auckland, New Zealand. The overall carriage rate for N. lactamica was 10.5% (95% CI 7.4-13.5%) with a peak carriage rate in 2-year-olds of 61.5% (95% CI 26.6-88.1%). Factors associated with a significant (P < 0.05) increase in the likelihood of carriage included runny nose, the number of people per bedroom and youth. Genetic analysis of isolates revealed a striking correlation of strains within the same household but a high level of diversity between households, suggesting that household contact is an important factor in acquisition. For household contacts aged less than 5 years, there was a higher rate of carriage amongst those in contact with MCD cases under 8 years old than for contacts of cases aged 8 years and over. It is likely that development of MCD is a reflection of the nature and intensity of the exposure to a virulent strain of N. meningitidis, coupled with an absence of host resistance among those individuals not carrying N. lactamica. PMID- 11057966 TI - Evidence of increased carriage of Corynebacterium spp. in healthy individuals with low antibody titres against diphtheria toxoid. AB - This study evaluated whether a correlation exists between carriage of corynebacteria and the lack of immunity to diphtheria toxoid. Samples of both nasal and pharyngeal secretions were taken from 500 apparently healthy subjects of both sexes and of all ages and inoculated onto Tinsdale's medium. A serum sample was also taken for ELISA test to determine the titre of diphtheria toxin antibodies. None of the subjects carried Corynebacterium diphtheriae. Ninety three strains of Corynebacterium spp. were isolated from 93 subjects and 86 of these were classified to species or group level by biochemical tests. C. xerosis was the most common (25.8%) followed by C. pseudodiphthericum (16.1%), C. jeikeium and C. striatum (both 10.8%), and C. urealyticum (9.7%). Three other species accounted for approximately 20% of strains and seven were unclassified as biochemically atypical corynebacteria. Non-protective antibodies to diphtheria toxin were found in 80 of the 93 subjects and a strong statistical association was demonstrated between carriage of corynebacteria and non-protective levels of anti-toxin antibodies. The remaining 13 subjects had protective levels of antitoxin antibodies. In contrast, only 45 of the 407 non-colonized subjects had non-protective antitoxin titres. The prevalence of carriage increased with age among males as did the percentage of non-protected subjects. The prevalence of female carriers of corynebacteria was significantly lower. Serum samples from 12 subjects with different antibody titres to diphtheria toxoid reacted to varying degrees with whole-cell lysates of a number of species of corynebacteria. The results suggest that a causal relationship may exist between nasopharyngeal carriage of corynebacteria and a low anti-diphtheria toxin immune response. PMID- 11057967 TI - The sero-epidemiology of diphtheria in Western Europe. ESEN Project. European Sero-Epidemiology Network. AB - Seven countries in Western Europe collected large, representative serum banks across the entire age range and tested them for diphtheria anti-toxin (sample size ranged from 2991 to 7715). Although a variety of assays were used, the results were all standardized to those of a reference laboratory and expressed in international units. The standardization process, and the availability of similar, large data sets allowed comparative analyses to be performed in which a high degree of confidence could be ascribed to observed epidemiological differences. The results showed that there were large differences in the proportion of adults with insufficient levels of protection amongst different countries. For instance, roughly 35% of 50- to 60-year-olds were found to be seronegative (titre < or = 0.01 IU/ml) in Finland compared with 70-75% in the United Kingdom. Furthermore, the proportion of seronegative adults would be expected to increase in some countries, notably Italy and the western part of Germany. In those countries with vaccination of military recruits there was a marked sex-related difference in the proportion of seropositive individuals. All countries have high levels of infant vaccine coverage (> 90%) but the accelerated schedule in the United Kingdom appears to result in lower anti-toxin titres than elsewhere. In Sweden, booster doses are not offered until 10 years of age which results in large numbers of children with inadequate levels of protection. Although the United Kingdom and Sweden both have higher proportions of seronegative children than elsewhere the likelihood of a resurgence of diphtheria in these countries seems remote. PMID- 11057968 TI - The European Sero-Epidemiology Network: standardizing the enzyme immunoassay results for measles, mumps and rubella. AB - The ESEN (European Sero-Epidemiology Network) project was established to harmonize the seroepidemiology of five vaccine preventable infections including measles, mumps and rubella in eight European countries. This involved achieving comparability both in the assay results from testing in different centres and also sampling methodology. Standardization of enzyme immunoassay results was achieved through the development of common panels of sera by designated reference centres. The panels were tested at the reference laboratory and then distributed to each participating laboratory for testing using their routine methods. Standardization equations were calculated by regressing the quantitative results against those of the reference laboratory. Our study found large differences in unitage between participants, despite all using an EIA method standardized against an international or local standard. Moreover, our methodology adjusted for this difference. These standardization equations will be used to convert the results of main serosurvey testing into the reference country unitage to ensure inter-country comparability. PMID- 11057969 TI - Regional differences in presentation of AIDS in Europe. AB - Data were collected on 6578 patients diagnosed with AIDS at 52 clinical centres in 17 European countries during an 1-year period from 1979 to 1989. The centres were divided into four regions, North, Central, Southeast, and Southwest. Differences in the incidence of most AIDS-defining opportunistic infections and malignancies were found. After adjusting for known possible confounders, statistically significant differences between regions remained. Pneumocystis carinii pneumonia (PCP) was more common in Northern Europe, Kaposi's sarcoma and toxoplasmosis in Central Europe, cytomegalovirus retinitis in South-eastern Europe, and extrapulmonary tuberculosis in South-western Europe. These differences we attribute primarily to different degrees of exposure to the respective underlying pathogens. The prevalence of these and other micro organisms will determine the clinical course of HIV infections in parts of Eastern Europe and elsewhere where the virus now is spreading. PMID- 11057970 TI - HIV incidence rates among drug users in northern Thailand, 1993-7. AB - Drug use is a major mode of HIV transmission in Thailand. This study determined HIV incidence rates among drug users in a regional drug treatment centre in northern Thailand. A retrospective cohort of repeatedly-hospitalized drug users between 1993 and 1997 was formed and HIV incidence rates were calculated. The overall incidence was 11.44 per 100 person-years of observation. Gender, age, religion, ethnicity, education, employment, income, reasons for drug use, type of drugs, mode of use, spending on drugs, and referral for treatment are associated with HIV incidence. However, there are no associations between HIV incidence and history of treatment and mode of discharge from the centre. This implies that current treatment modality has no impact on HIV infection risk and other therapeutic approaches should be explored. PMID- 11057971 TI - Seroprevalence of HIV and HTLV in a representative sample of the Spanish population. AB - HIV and HTLV seroprevalence was determined by means of unlinked anonymous testing of 2144 sera, originally obtained from primary care patients by representative sampling of the Spanish population aged 15-39 years in 1996. HIV-1 seroprevalence was 4.3 per 1000 population in the 15-39 years age group [95% confidence interval (CI), 1.5-10.7] and 5.6 per 1000 (95% CI, 1.8-15.3) in the 20-39 years age group. Seroprevalence proved higher in males and urban residents. No antibodies to HIV-2 and HTLV-I were detected in any of the sera studied. However, presence of antibodies to HTLV-II was confirmed in one serum sample, while HTLV seroreactivity, though detected in another, could not be typed. The two HTLV positive results equated to a seroprevalence of 1.9 per 1000 in the 20-39 years age group (95% CI, 0.3-8.6). HIV-I seroprevalence was consistent with previous estimates yielded by back-calculation. The level of HTLV seroprevalence found suggests endemicity. PMID- 11057972 TI - HBV, HCV and HDV infections in Albanian refugees in Southern Italy (Apulia region). AB - The seroprevalence of hepatitis B, C and D markers was assessed in a sample of 670 Albanian refugees in Southern Italy in 1997. The mean age was 25 years (S.D. = 12.3). Of study subjects 62.1% (95% CI: 58.4-65.7) were positive for anti-HBc antibodies and 13.6% (95% CI: 10.9-16.1) for HBsAg. The prevalence of anti-HBs was 47.6% (95% CI: 43.8-51.3). Among HBsAg carriers the prevalence of HBeAg was 7.7% (95% CI: 2.2-13.1). The highest carrier rate for HBsAg (25.5%; 95% CI: 16.7 34.3) was found in the age group 21-25 years. A relevant finding was a prevalence of HBsAg of 8.1% in children 10 years and under. The prevalence of anti-HCV antibodies was 0.3% (95% CI: 0.0-0.7) while only one of the HBsAg carriers was positive for anti-HDV (1.1%, 95% CI: 0-3.2). In Albania, hepatitis B infection represents a public health priority that should be addressed by a universal vaccination campaign. PMID- 11057973 TI - High circulation of hepatitis B virus (HBV) precore mutants in Tunisia, North Africa. AB - Hepatitis B Virus (HBV) e antigen (HBeAg), HBV DNA and precore mutations affecting HBeAg expression during active replication were studied in 72 Tunisian hepatitis B surface antigen (HBsAg) positive individuals: 30 asymptomatic carriers of the virus, 37 with chronic hepatitis and 5 with acute hepatitis. HBV DNA was detected in 44 patients, but only 20% of them expressed HBeAg. Precore mutant strains, with mutations at position 1896 or at positions 1896 and 1899, were detected by PCR-hybridization in 86 and 36% of patients, respectively. Wild type strains were detected in 54% of patients. Precore mutants were found in chronically and in acutely infected individuals, in patients with severe and asymptomatic infections, in HBeAg positive as well as HBeAg negative individuals. These results show the high frequency of HBV precore mutants in Tunisia. PMID- 11057974 TI - Human rabies in Lebanon: lessons for control. AB - Human rabies is known to be endemic in countries bordering Lebanon, but its prevalence in Lebanon has not been studied before. All eight cases of human rabies reported to the Lebanese Ministry of Public Health between 1991 and 1999 were reviewed, as well as three other cases admitted to the American University of Beirut Medical Center. A total of 1102 cases of animal bites to humans, the majority of which were dog bites, were reported to the Ministry of Public Health between 1991 and 1996. In this period, 2487 doses of rabies vaccine were administered to the above group, as post-exposure prophylaxis. Veterinarians, a high risk and educated group, were interviewed, and only 7 out of 72 were found to have been vaccinated. Major improvements in surveillance and reporting, better control of animal rabies, more awareness especially among high risk groups, and regional cooperation, are all needed to prevent and control this deadly infection. PMID- 11057975 TI - Epidemiological features of and public health response to a St. Louis encephalitis epidemic in Florida, 1990-1. AB - A St. Louis encephalitis (SLE) epidemic in Florida during 25 weeks in 1990-1, resulted in 222 laboratory-diagnosed cases, an attack rate in the 28 affected counties of 2.25/100,000. Disease risk rose with advanced age, to 17.14/100,000 in persons over 80 years, and all 14 fatal cases were in persons over 55 years (median, 70 years). Community serosurveys in Indian River County, the epicenter of the outbreak (attack rate 21/100,000), showed acute asymptomatic infections in 3.6% of the persons surveyed, with higher rates in persons with outdoor occupational exposure (7.4%) and in clients of a shelter for the indigent (13.3%). A matched case-control study found that evening outdoor exposure for more than 2 h was associated with an increased risk for acquiring illness (odds ratio [OR] 4.33, 95% CI 1.23-15.21) while a number of recommended personal protective measures were protective. Four SLE patients were dually infected with Highlands J virus, the first reported cases of acute infection with this alphavirus. The case-control study provided the first evidence that a public education campaign to reduce exposure had a protective effect against acquiring the disease. PMID- 11057976 TI - Surveillance of dengue fever in French Guiana by monitoring the results of negative malaria diagnoses. AB - Surveillance of dengue fever is mainly based on specific laboratory tests. However non-specific systems, such as clinical surveillance, are also required. In French Guiana, we have tested a non-specific laboratory surveillance system where different biological examinations performed for other reasons than the diagnosis of dengue fever were analysed as methods for dengue fever surveillance. The number of negative malaria diagnoses in Cayenne and Kourou was found to be the best indicator of dengue fever infections in these towns. This surveillance system appears to be very simple and reliable, and a test which could serve as an indicator that is likely to be found everywhere. PMID- 11057977 TI - Silent spread of dengue and dengue haemorrhagic fever to Coimbatore and Erode districts in Tamil Nadu, India, 1998: need for effective surveillance to monitor and control the disease. AB - Dengue fever (DF) or dengue haemorrhagic fever (DHF) has not previously been reported in Coimbatore and Erode districts in Tamil Nadu in India. In 1998, 20 hospitalized cases of fever tested positive for dengue virus IgM and/or IgG antibodies. All of them had dengue-compatible illness, and at least four had DHF. Two of them died. Sixteen cases were below 10 years of age. The cases were scattered in 15 distantly located villages and 5 urban localities that had a high Aedes aegypti population. Although the incidence of dengue-like illness has not increased recently, almost 89% (95/107) of samples from healthy persons in the community tested positive for dengue IgG antibodies. The study showed that dengue has been endemic in the area, but was not suspected earlier. A strong laboratory based surveillance system is essential to monitor and control DF/DHF. PMID- 11057978 TI - Epidemiologic features of 65 Creutzfeldt-Jakob disease patients with a history of cadaveric dura mater transplantation in Japan. AB - A total of 65 cases of Creutzfeldt-Jakob disease with a history of cadaveric dura transplantation in Japan were analysed to clarify the epidemiologic features of such patients and to explore whether other such patients will appear in the future. The age at transplantation averaged 44.4 years with a standard deviation of 14.4 years. The age at onset had an average of 53.0 years with a standard deviation of 14.1 years. The shortest latent period was 14 months, and the longest was 218 months with an average of 103.1 months and a standard deviation of 49.9 months. From the relationship between the calendar year at transplantation and the latent period, other such patients will appear in the near future. The current data suggested that several patients with Creutzfeldt Jakob disease will occur from those receiving cadaveric dura mater grafts in the near future. PMID- 11057979 TI - Transmission of the nocturnal periodic strain of Wuchereria bancrofti by Culex quinquefasciatus: establishing the potential for urban filariasis in Thailand. AB - Control programmes have reduced the prevalence of Bancroftian filariasis in Thailand to low levels. Recently, there has been an influx of more than one million Myanmar immigrants into urban centres of Thailand. The prevalence of patent Wuchereria bancrofti infection in these immigrants (2-5%) has prompted concern in the public health community that the potential now exists for a re emergence of Bancroftian filariasis in Thailand. It is possible that an urban cycle of transmission could become established. The Myanmar immigrants are infected with the nocturnal periodic (urban) type W. bancrofti for which Culex quinquefasciatus serves as the main vector. The Thai strains of Cx. quinquefasciatus have never been reported to transmit Bancroftian filariasis. Our results of feeding experiments demonstrated that the Thai Cx. quinquefasciatus are permissive for the development of Myanmar W. bancrofti to infective third stage larvae thus establishing the potential for establishing an urban cycle of transmission in Thailand. We also adapted the SspI repeat PCR assay for the identification of infective mosquitoes that was capable of detecting a single infective stage larvae in a pool of 100 mosquitoes. PMID- 11057980 TI - Existence of two geographically-linked clonal lineages in the bacterial fish pathogen Photobacterium damselae subsp. piscicida evidenced by random amplified polymorphic DNA analysis. AB - In this work, we applied the random amplified polymorphic DNA (RAPD) technique to evaluate the genetic diversity in Photobacterium damselae subsp. piscicida (formerly Pasteurella piscicida), an important pathogen for different marine fish. Regardless of the oligonucleotide primer employed, the 29 isolates of Ph. damselae subsp. piscicida tested were separated into two groups, the RAPD-PCR analysis differentiated the European strains from the Japanese strains. The similarity between both groups estimated on the basis of the Dice coefficient was 75-80%. These results show that European and Japanese isolates of Ph. damselae subsp. piscicida, regardless of their host fish species, belong to two different clonal lineages. Our findings also indicate that RAPD profiling constitutes a useful tool for epidemiological studies of this fish pathogen. PMID- 11057982 TI - Does more equal less or does less equal more? PMID- 11057981 TI - A comparison of Escherichia coli O157 isolates from cattle in Japan and the USA by molecular biological methods. AB - Escherichia coli O157 isolates from cattle in Japan (n = 91) and in the USA (n = 415) were compared by pulsed-field gel electrophoresis of endonuclease-cleaved genomic DNA, location of the stx genes and bacteriophage typing. Three isolates from cattle in Japan with high similarity to isolates from cattle in the USA were found. Isolates from cattle farms in Japan and the USA may share a common source. PMID- 11057983 TI - An audit of open and laparoscopic inguinal hernia repair. AB - A retrospective case review to describe current practices and outcomes of patients undergoing inguinal hernia repair was undertaken in a principal referral hospital (John Hunter Hospital, Newcastle, New South Wales). The participants were patients who had elective laparoscopic or open inguinal hernia repair between 1 June 1997 and 31 May 1998. The main outcome measures were duration of surgery, type of anaesthesia, use of antibiotics, length of stay, postoperative complications, analgesic administration, and costs. Laparoscopic repair was advantageous in terms of length of stay and use of analgesia, but at a higher in theatre financial cost. Low rates of day stay surgery and use of local anaesthesia, and a high rate of antibiotic administration were apparent. It was concluded that local surgical practice has adopted some but not all advances described in the literature. Performance of a more expensive hernia repair in some patients may mitigate against performance of any repair in others. PMID- 11057984 TI - The evaluation of wound infection after arterial surgery. AB - Many of the wound infections that occur after arterial surgery require prolonged attention with frequent changes of dressings. However, the traditional way of reporting wound infections is to only declare the overall incidence of this adverse event. In this study, we have evaluated 302 patients undergoing arterial surgery to determine the relationship between the incidence of wound infection, the length of time that wounds were dressed, and the results of a modified ASEPSIS wound scoring system. The overall incidence of wound infection was 14% (43/302). It was noted that patients with high wound scores had a correspondingly high incidence of conventionally defined wound infections; and, in addition, such patients required dressing changes over prolonged periods. It is concluded that the documentation of both the duration of wound dressings and a wound scoring system are useful tools when evaluating the outcome of patients after arterial surgery. PMID- 11057985 TI - Translating quality into research: do we need more research into quality or should quality activities be conducted using the principles and methodological rigour of scientific research? Australasian Association for Quality in Health Care. PMID- 11057986 TI - Home rehabilitation for older adults with fractured hips: how many will take part? AB - Rehabilitation at home is a new 'technology' which has been promoted as an efficient alternative to hospital rehabilitation for older patients with conditions such as fractured hip. In Australia, no formal description of elderly patients with fractured hips likely to be eligible for home rehabilitation has been made and the acceptability of such services is unclear. Using information obtained prospectively from a consecutive sample of 188 patients with a fractured hip we describe the characteristics of older adults who were eligible for a trial examining home versus hospital rehabilitation. While staff assessed 36% of patients as eligible, only 20% were both eligible and agreeable. Reasons for refusal to participate included a preference for inpatient rehabilitation (26%), family reluctance (26%) and anxiety regarding the ability to manage at home (16%). Our results suggest that home rehabilitation is suitable for the least disabled group but is still unacceptable to many elderly patients and their families. As the population ages and hip fractures increase, home rehabilitation in its current form will have little impact on future bed needs. PMID- 11057987 TI - An hypothesis paper on practice environment and the provision of health care: could hospital occupancy rates effect quality? AB - This paper will explore whether hospital occupancy influences quality of care. It discusses the 'systems' theory of error causation in the context of adverse medical outcomes. It then relates how high occupancy rates may cause problems in hospital systems. The evidence relating to quality of care and occupancy is reviewed. Finally, a new method of studying this relationship using time series analysis is proposed. We conclude that the relationship requires further exploration since revealing 'system' problems may compel clinicians to expose problems medical errors. PMID- 11057988 TI - The quality of medical laboratory practice in Trinidad and Tobago, West Indies. AB - The introduction of automated chemical analyzers in the laboratory service has the potential of adversely affecting professionalism in laboratory practice. The present study assesses the quality of medical laboratory technicians in Trinidad and Tobago using structured questionnaires. Some of the critical questions included job status, years of experience, training, qualification(s) and knowledge of quality assurance and its application. About 82% of laboratory technicians responded to the study. The majority of technicians (62%) had diploma certificates while only one (1.2%) had a postgraduate degree. Although the majority (91.7%) of technicians knew about quality assurance, 36% learnt on the job and 59% knew they were not professionally trained. The results showed that there is paucity of highly trained laboratory technicians in Trinidad and Tobago and this has significant implications on the technical initiative and quality of medical laboratory practice in this country. We recommend the establishment of appropriate professional institutions for training medical laboratory technologists and regular expert inspection and accreditation of all medical laboratories in the country. PMID- 11057989 TI - Institute for Health Care Improvement Collaborative Trial to improve process times in an Australian emergency department. AB - This study describes an Australian emergency department's (ED) experience with a quality improvement methodology from the USA. The Institute for Health Care Improvement (IHI) conducts collaboratives between clinical groups with similar interests, in this case ED. Their quality improvement model is described. Our involvement with the IHI showed the model to be transferable outside the USA. In applying the model to operational and clinical projects we were successful in meeting our goals to reduce clinical times: for time to analgesia (P= 0.34), time to thrombolysis (P= 0.30) and time to antibiotics in neutropenic patients (P= 0.015). We were unable to reach statistical significance in improvements due to the small sample sizes and sampling techniques. Changes in operational times were not clinically significant but almost reached statistical significance (e.g. median total length of stay in the ED fell 4 min (P= 0.06)). The near statistical significance of a small change was due to the large numbers of patients sampled. PMID- 11057990 TI - Quality assurance and technology assessment: pieces of a larger puzzle. AB - Increasing integration of health care and health services research has resulted in an overlap between disciplines involved in the evaluation of clinical practice. We have examined the relationships of quality assurance (QA), medical technology assessment (TA), clinical epidemiology (CE) and evidence-based medicine (EBM) from an historical perspective. Clinicians, patients and administrators need local information on effectiveness of routine care. Information from trials alone, efficacy data, will not suffice nor can it be culled from administrative databases designed for other purposes. The current activities of QA should be therefore be expanded to include the study of the effectiveness of interventions in terms of appropriateness of use, patient outcomes and study of the determinants of outcomes, as seen from the perspective of doctors, patients, administrators and policy makers, using data collected during the course of routine patient care. With the assistance of information technology, with methodological support and multidisciplinary cooperation, clinicians can do this as part of a more broadly defined clinical research. Quality assurance and TA both evolved with the objective of studying clinical care but have quite different historical roots, complementary perspectives and objectives, use different methods and involve a different set of practitioners. Quality assurance is a type of 'formative' evaluation conducted in the clinical setting using indicators as flags of process or outcome events of interest, simple surveys and audit studies. Its primary aim is to achieve incremental improvement rather than to simply pass judgement. An important underlying assumption is that health care behaves as a complex dynamic system. Technology assessment, a form of summative evaluation with an orientation towards policy, synthesises information from formal scientific studies of efficacy in the form of clinical trials and studies of cost-effectiveness. For the evaluation of the impact of any technology more complex than a drug, the complementary contributions of both of these disciplines is needed, and QA and TA should work cooperatively in tandem with the support of CE and EBM. PMID- 11057991 TI - The appropriateness of admissions and the influences on a decision to admit. AB - Within health services there is concern that escalating costs may reflect an inappropriate use of services and that services should avoid admitting patients who do not require acute care. The problem is, if these patients exist, how may they be identified and is this the sole consideration in patient management? This triangulated study examines the appropriateness of admissions, factors that influence that admission, and the potential to develop alternate models of care for medical patients. A utilization review, auditing 521 medical patients, provided a percentage of appropriateness of admission and stay, identified the barriers to discharge and those patients suitable for programs such as hospital in the home. Complementing the review, interviews with eight physicians explored how they decided to admit patients, what factors influenced that decision and their attitude to the development of these models of care. PMID- 11057992 TI - Establishing adolescent and young adult therapy programs. AB - Perth Clinic is a 35-bed private psychiatric clinic. In February 1999 an adolescent and young adult therapy service was introduced in response to an increasing number of enquiries regarding the treatment of adolescent and young adults with psychiatric disorders. This program was developed by a working party of mental health professionals including psychiatrists, clinical psychologists, nurses and occupational therapists. The program design described the type of program to be delivered, its content and structure, and the method of evaluation. In consultation with similar services around the world, Perth Clinic developed two therapy streams for this population. The first is an inpatient program based on an open group format of 4 1/2 hours of therapy daily. The second is a closed group program conducted for two 3 hour sessions per week. Groups are held in the late afternoon (16.00-19.00 h) for an 8 week period. Both streams have a maximum of eight participants and all participants are referred by a psychiatrist. Outcomes are measured by a battery of clinical questionnaires which provide objective measurement of mood changes. Patient and parent satisfaction with the program is also measured. Indicators at this early stage are that this is an effective intervention program with high rates of attendance and good prognostic data. This study describes the first 6 months of this program's operation. PMID- 11057993 TI - Marycrest strength training group. AB - The aim of the Marycrest Strength Training Group was to provide the frail elderly residents of the Marycrest Retirement Centre, Kangaroo Point, Queensland, with the opportunity to improve their muscle strength and functional abilities by participating in a supervised, safe and enjoyable group activity. PMID- 11057994 TI - Cerebrovascular accident clinical pathway. AB - The cerebrovascular accident (CVA) clinical pathway project was selected to complement the work already underway within the West Moreton Health Services District such as the development of a continuum of care model, revision of work practices to complement the new hospital redevelopment and encouraging team and evidence-based approaches to problem solving. Specific objectives were set for the project along with a detailed evaluation plan. A steering group was convened to run the project and a full time project officer was appointed. At the end of the 12 month period all the objectives were met. Specific achievements included a reduction in the overall average length of stay for those patients who experience CVA, improved clinical outcomes and a more effective use of resources. Quality of care has been improved through the preparation of specialized clinical pathway documentation, education packages, patient surveys, focus groups, independent reviews and benchmarking. Complementing these measures has been a series of process changes and environmental modifications. Furthermore, good working relationships have been established with private sector providers of health care and other external bodies. The development of the CVA clinical pathway at the Ipswich Hospital has meant timely referrals and a streamlined assessment and referral process to get patients into rehabilitation sooner. It has promoted good communication between, and recognition of, the professional roles of various team members and has put the patient back at the centre of the care process. PMID- 11057995 TI - Extended paediatrics: acute care in children's homes. AB - The Extended Paediatric Service was established in 1996 to provide acute care services to children in their homes with the aim of reducing the number of hospital admissions for children and shortening hospital stays while maintaining continuity of care. Extended Paediatrics builds on, and extends, the principles of Discharge Liaison and Hospital In The Home programmes. The service is staffed by nurses who also work within the paediatric ward of the Launceston General Hospital, providing a high degree of continuity of care for these children. Children are admitted to the service through consultation between medical staff, nursing staff and the child's family. Between August 1996 and December 1999 a total of over 1200 visits to 48 children have been made for various treatments, for tests, education or follow up. Improved outcomes for children and their families have been reported by both staff and parents as a result of the service. PMID- 11057996 TI - Avulsion injuries to the brachial plexus and the value of motor reinnervation by ipsilateral nerve transfer. PMID- 11057997 TI - Transfer of a single fascicle from the ulnar nerve to the biceps muscle after avulsions of upper roots of the brachial plexus. AB - Thirty-six patients with avulsions of upper roots of the brachial plexus underwent transfer of a single fascicle from the ulnar nerve to the proximal motor branch of the biceps muscle to restore elbow flexion. The mean period of follow-up was 22 months. The average reinnervation time for the biceps muscle was 3.3 months. Thirty-four patients achieved biceps strength of Medical Research Council grade 3 or better. The operative results in the patients with C5, C6 avulsions were better than those with C5, C6, C7 avulsions. At the last follow-up examination, grip strength, pinch strength, moving two-point discrimination and the strength of flexion of the wrist on the affected side was not worse than before surgery in any patient. PMID- 11057998 TI - Nerve transfer to the median nerve using parts of the ulnar and radial nerves in the rabbit--effects on motor recovery of the median nerve and donor nerve morbidity. AB - In this study, motor re-innervation of the median nerve by transfer of one-third, one-half, and two-thirds of either the agonistic ulnar nerve or the antagonistic radial nerve was investigated in both extremities of 20 rabbits. Recipient median nerve: Muscle contraction force of the flexor digitorum sublimus muscle after a one-third and a one-half of the ulnar nerve transfer achieved an average of 75 and 97% muscle power respectively as compared to conventional end-to-end neurorrhaphy. Muscle contraction force after one-third or one-half of the radial nerve transfer was significantly lower (36%). Donor nerves: Extensor carpi radialis muscle or flexor carpi ulnaris muscle contraction force 6 months postoperatively demonstrated a significant decrease after a one-half ulnar nerve and a two-thirds ulnar or radial nerve transfer, but not after a one-third transfer of either radial or ulnar nerves. Histologically, the number of axons in the re-innervated median nerve and both donor nerves distal to the coaptation site seemed to follow variable patterns. It was concluded that in the rabbit use of one-third of the agonistic ulnar nerve for re-innervation of the median nerve results in useful motor recovery with negligible donor site morbidity. Clinically, this technique may offer an alternative option for proximal nerve injuries or for free functioning muscle transplantations. PMID- 11057999 TI - Brachial plexus injuries in scapulothoracic dissociation. AB - The rare condition of scapulothoracic dissociation (STD) is characterized by a lateral displacement of the scapula from the thoracic cage following severe trauma to the scapular girdle. This study presents an analysis of five STDs. There were three supraclavicular brachial plexus palsies and two retro- and infraclavicular palsies. Recovery of elbow flexion was obtained in only two cases. Nerve damage dominates the prognosis and nerve recovery only rarely occurs. Nerve surgery should attempt to reestablish elbow flexion. PMID- 11058000 TI - Midcarpal arthrodesis with complete scaphoid excision and interposition bone graft in the treatment of advanced carpal collapse (SNAC/SLAC wrist): operative technique and outcome assessment. AB - Thirty-six patients with stage II or III SNAC and SLAC wrists were treated by midcarpal arthrodesis and complete scaphoid excision. When assessed at a mean follow-up of 25 months, pain was significantly reduced both under resting and stress conditions. The active range of motion was 54% of the contralateral wrist and grip strength was 65% of the non-operated hand. The mean DASH score was 28 points, the Mayo wrist score was 63 points, and the Krimmer wrist score was 68. Correlation of the wrist scores with the DASH values demonstrated a significant correlation. Our data demonstrate that midcarpal fusion with complete excision of the scaphoid is a reliable procedure for treating advanced carpal collapse. PMID- 11058001 TI - Pressure mapping of the radioulnar carpal joint: effects of ulnar lengthening and wrist position. AB - The purpose of this study was to investigate the effects of ulnar lengthening and wrist position on force transmission through the radioulnar carpal joint in a forced dorsiflexed wrist position. Eight cadaveric arms were subjected to a 30 kg compressive load directed down the forearm towards the wrist. A pressure sensor recorded forces across the wrist joint with the triangular fibrocartilage complex intact and excised. The biomechanics of the distal radioulnar joint were altered by changes in ulnar length and wrist position. Pressures at the ulnolunate articulation increased as the ulna was lengthened and were significantly lower when the triangular fibrocartilage complex was excised. An inverse relationship between triangular fibrocartilage complex thickness and ulnar variance was shown. Greater increases in ulnolunate pressure were observed in more positive ulnar variant wrists. PMID- 11058002 TI - A comparison of trapeziectomy with and without ligament reconstruction and tendon interposition. AB - Forty-three patients were randomly allocated to undergo either trapeziectomy alone (control) or with a ligament reconstruction and tendon interposition (LRTI) using an abductor pollicis longus tendon slip. The patients were reviewed at a median 13 (range, 7-29) months after surgery. The demographic characteristics, severity of disease and pre-operative clinical measurements of the two study groups were indistinguishable but LRTI lengthened the operation by approximately 15 minutes. Both groups expressed equal satisfaction with the operation and there were no significant differences between the two treatment groups. Simple trapeziectomy is an effective operation for osteoarthrosis at the base of the thumb and the addition of a ligament reconstruction was not shown to confer any additional benefit. PMID- 11058003 TI - Carpal tunnel release using a short palmar incision and a new knife. AB - A new knife with its own battery powered light source (Knifelight, Stryker Instruments, Kalamazoo, Michigan, USA) was used for carpal tunnel release in 31 wrists of 25 patients. Under local anaesthesia, a short palmar incision was used and the carpal tunnel contents were visualized during division of the flexor retinaculum. The mean operation time was 11 minutes and no major complications were seen. The patients could use their hands for self-care after 3 days and returned to work at a mean of 23 days. At a minimum follow-up of 6 months, all but one of the patients were satisfied with the final result. Mild scar tenderness was seen in two patients and pillar pain in one patient. PMID- 11058004 TI - Value of electrodiagnostic tests in carpal tunnel syndrome. AB - Eighty-five patients who were treated surgically for carpal tunnel syndrome were reviewed by a prospective study to assess the relationship between electrodiagnostic tests and clinical outcome. Before surgery all patients completed a self-administered questionnaire, and an electrophysiological examination was done to assess distal motor latency, sensory and motor amplitudes, and sensory nerve conduction velocity. After 6 months follow-up all these tests were repeated. There was significant correlation between improvement in sensory nerve conduction velocity and clinical improvement at follow-up. PMID- 11058005 TI - Elbow extension and flexion-supination deformities in tetraplegia. AB - This paper demonstrates the flexion-supination and extension deformities of the elbows in complete tetraplegics and describes the outcome of simple treatments. The principal conclusion is that these two deformities are easily corrected by very simple methods. After treatment the patients could fully extend and flex their elbows and fully supinate and pronate their forearms. One of 11 patients, whose bilateral deformities kept recurring, was a failure. PMID- 11058006 TI - Ultrasound for diagnosis of scaphoid fractures. AB - We investigated the diagnostic value of two different ultrasound scanning methods for the early diagnosis of acute scaphoid fractures. Fifty-seven patients with ten scaphoid fractures were assessed within a week of injury. The accuracy of the ultrasound assessment was 84% and its specificity was 91%. However, its sensitivity was only 50%. We conclude that ultrasound examination is unreliable for the diagnosis of acute scaphoid fractures. PMID- 11058007 TI - Rapid repeat testing of grip strength for detection of faked hand weakness. AB - This study assessed the use of rapid, repeated measurement of grip strength to detect feigned hand weakness. Normal participants, performing with maximum effort or feigning hand weakness, and patients recovering from carpal tunnel surgery were asked to grip a Jamar dynamometer alternately with each hand on ten occasions. The results showed that grip strength fatigued by an average of 23% during the test in the normal participants, 18% in participants faking weakness, and increased by 2% in the carpal tunnel decompression patients. An increase in grip strength after the first effort was found in 39% of normal participants, 52% of participants faking hand weakness and in 69% of the carpal tunnel decompression patients. These results suggest that rapid, repeated measurement of grip strength is not a reliable discriminator of true and faked hand weakness. PMID- 11058008 TI - An experience of the Snow-Littler procedure. AB - This paper reviews the results of the Snow-Littler procedure performed in twelve hands with classical central longitudinal deficiency and in one hand with symbrachydactyly, cleft type. There were no instances of major flap necrosis although two flaps showed tip ischaemia. The width of the first web was, in the main, satisfactory but four webspace revisions were performed. Supplementary skin grafting at the time of surgery was necessary in complete and/or complex thumb index syndactylies and in the patient with symbrachydactyly. In eight cases, a transverse metacarpal ligament was reconstructed. In the five other cases, no clinical instability or radiological divergence of the index and ring fingers occurred, in spite of no transverse metacarpal ligament reconstruction. Three de rotational osteotomies of transposed index fingers were performed in patients who had a transverse metacarpal ligament reconstruction. These results indicated significantly improved appearance and improved function following the Snow Littler procedure. PMID- 11058009 TI - The donor foot in free toe or joint transfers. AB - Eighty-four cases of free second toe, multiple toe or second metatarsophalangeal joint transfers were studied by case review and follow-up. The function of the donor foot had recovered completely within 6 months in 89% of patients. The wounds on the donor foot healed in 2-3 weeks in 90% of patients. Slight numbness on the dorsal aspect of the donor foot, intolerance to cold, mild reduction in push-off, scar tenderness and pain or swelling occurred in only a few patients and generally were not considered of significance. Multiple toe transfers created more donor problems in terms of healing and appearance. Some foot deformities with plantar callosities were observed at long term review. However, all patients were capable of work and normal activities. PMID- 11058010 TI - Reverse radial forearm fascial flap for soft tissue coverage of hand and forearm wounds. AB - Six patients with severe hand and forearm injuries involving open wounds and exposed structures were treated with reverse radial forearm fascial flaps and split-thickness skin grafts for soft tissue cover. There were five men and one woman aged between 16 and 36 years. Injuries included soft tissue avulsion on the dorsum of the hand and fingers, extensive flexor and extensor tendon damage, multiple phalangeal fractures, a grade IIIB open dislocation of the index to little carpometacarpal joints, a grade III open metacarpal fracture and a finger amputation. The average wound size was 9 cm in length and 7 cm in width. The mean duration of follow-up was 12 months (range, 5-20 months). All flaps healed well, and all patients were satisfied. PMID- 11058011 TI - A new core suture technique for flexor tendon repair: biomechanical analysis of tensile strength and gap formation. AB - The purpose of this study was to test in vitro a new flexor tendon suture technique that has been developed to withstand the in vivo forces of active tendon motion. This new core suture technique involves locking loops in the form of a cross stitch. The new technique was tested for ultimate tensile strength and gap formation in cadaver human flexor tendons on a tensile testing machine. The new technique proved significantly stronger than the modified Kessler core suture. PMID- 11058012 TI - History of the Deutschsprachige Arbeitsgemeinschaft fur Handchirurgie and the Deutsche Gesellschaft fur Handchirurgie. PMID- 11058013 TI - Madelung deformity treated with Ilizarov technique: a report of two cases. AB - Two children with painful and progressive Madelung deformities were treated by osteotomy of the radius and subsequent angular correction and bone lengthening using the Ilizarov technique. Both children were radiologically improved and free of pain at follow-up. PMID- 11058014 TI - Variable expression of isolated familial long-ring-little syndactyly. AB - A family with isolated long-ring-little syndactyly is reported to demonstrate that the responsible autosomal gene may cause either simple or complex syndactyly. PMID- 11058015 TI - Dorsal dislocation of the capitate. AB - We report a case of dorsal dislocation of the capitate and third metacarpal, and discuss the possible classification of this pattern of injury. PMID- 11058016 TI - The use of silicone tubing in the late repair of the median and ulnar nerves in the forearm. PMID- 11058017 TI - Repair of the flexor pollicis longus tendon. PMID- 11058018 TI - MAFF announces further support measures as more CSF cases are confirmed. PMID- 11058019 TI - Knowledge and practice: the future of companion animal medicine? PMID- 11058020 TI - Temporal aspects of the epidemic of bovine spongiform encephalopathy in Great Britain: holding-associated risk factors for the disease. AB - The objectives of this study were first to describe the pattern of the epidemic of bovine spongiform encephalopathy (BSE) in Great Britain in terms of the temporal change in the proportion of all cattle holdings that had experienced at least one confirmed case of BSE to June 30, 1997, and secondly to identify risk factors that influenced the date of onset of a holding's first confirmed BSE case. The analyses were based on the population of British cattle at risk, derived from agricultural census data collected between 1986 and 1996, and the BSE case data collected up to June 30, 1997. The unit of interest was the cattle holding and included all those recorded at least once on annual agricultural censuses conducted between June 30, 1986, and June 30, 1996. The outcome of interest was the date on which clinical signs were recorded in a holding's first confirmed case of BSE, termed the BSE onset date. Univariate and multivariate survival analysis techniques were used to describe the temporal pattern of the epidemic. The BSE epidemic in Great Britain started in November 1986, with the majority of affected holdings having their BSE onset date after February 1992. After adjusting for the effect of the size and type of holding, holdings in the south of England (specifically those in the Eastern, South east and South west regions) had 2.22 to 2.43 (95 per cent confidence interval [CI] 2.07 to 2.58) times as great a monthly hazard of having a BSE index case as holdings in Scotland. After adjusting for the effect of region and type of holding, holdings with more than 53 adult cattle had 5.91 (95 per cent CI 5.62 to 6.21) times as great a monthly hazard of having a BSE index case as holdings with seven to 21 adult cattle. Dairy holdings had 3.06 (95 per cent CI 2.96 to 3.16) times as great a monthly hazard of having a BSE index case as beef suckler holdings. These analyses show that there were different rates of onset in different regions and in holdings of different sizes and types, that the epidemic was propagated most strongly in the south of the country, and that the growth of the epidemic followed essentially the same pattern in each region of the country, with modest temporal lags between them. The control measures imposed in 1988 and 1990 brought the expansion of the epidemic under control, although the rate of progress was slowed by those regions where the effectiveness of the control methods took some time to take full effect. PMID- 11058021 TI - Long-term outcome of surgery for dogs with cranial cruciate ligament deficiency. AB - Fifty-eight dogs with cranial cruciate ligament deficiency were assessed and treated surgically. At an average of 50 months postoperatively, the functional outcome was assessed by means of an owner-based clinical assessment and a clinical examination. Client-based data were available for 26 dogs and 20 dogs were reassessed after 50 months. The results were compared with the initial values and with data from an assessment 13 months postoperatively. The level of disability at 50 months was judged to be significantly less than initially. However, there were no differences between the initial assessments and those made after 50 months for the perceived 'effect of cold weather' and the dogs' 'ability to jump', despite both measures having improved after 13 months. Age and meniscal injury were identified as poor prognostic indicators for the long-term outcome. The equivalent joint on the contralateral limb deteriorated significantly during the study. PMID- 11058022 TI - Presence of bovine herpesvirus 1 gB-seropositive but gE-seronegative Dutch cattle with no apparent virus exposure. AB - Two hundred and thirty-seven of 2052 cattle which had not been vaccinated against bovine herpesvirus 1 (BHV-1) were seropositive in a glycoprotein B (gB)-blocking ELISA, but seronegative in a glycoprotein E (gE)-blocking ELISA. In order to detect whether they were latently infected with BHV-1, 10 of them were treated with corticosteroids in an attempt to reactivate putatively latent virus. After successive treatments with dexamethasone and prednisolone, no virus excretion was detected and they showed no increase in antibody titres. In contrast, one gE seropositive animal re-excreted BHV-1 and had a four-fold increase in antibody titre after the corticosteroid treatments. After slaughter, no BHV-1 DNA could be detected with a sensitive PCR in samples of the trigeminal, cervical and sacral ganglia and spinal cords of the gE-seronegative cattle. PMID- 11058023 TI - Some aspects of erythrocyte metabolism in a dog with polycythaemia vera. AB - An 11-year-old female crossbred dog showed signs of polyuria, polydipsia, vomiting, posterior weakness and ataxia. Clinical and laboratory findings suggested the diagnosis of polycythaemia vera. The haematological values shown over a six-month period are presented. In four samples some aspects of erythrocyte function (glucose-6-phosphate dehydrogenase [G6PD] and pyruvate kinase [PK] activities, 2,3 diphosphoglycerate [2,3 DPG] concentration, osmotic fragility and intracellular sodium and potassium concentrations) were studied. Variable activities of G6PD and PK, probably related to different reticulocyte number, were detected together with normal osmotic fragility and intracellular sodium and potassium concentrations. 2,3 DPG concentration was higher than normal in all four samples. This could be interpreted as a response to a low tissue perfusion rather than a higher content of 2,3 DPG in red blood cells from the polycythaemic dog. PMID- 11058024 TI - Mixed mycoplasma mastitis outbreak in a dairy herd. PMID- 11058025 TI - Malignant oedema in a guanaco (Lama guanicoe). PMID- 11058026 TI - Milk withdrawal periods. PMID- 11058027 TI - Fish and the Veterinary Surgeons Act. PMID- 11058028 TI - Dispensing review. PMID- 11058029 TI - Ingestion of equine moxidectin by dogs. PMID- 11058030 TI - RCVS annual retention fee. Royal College of Veterinary Surgeons. PMID- 11058031 TI - Assessment of stranded cetaceans. PMID- 11058032 TI - Coenurus serialis in a pet rabbit. PMID- 11058033 TI - Identification of HIV-1 integrase inhibitors via three-dimensional database searching using ASV and HIV-1 integrases as targets. AB - Integration of viral DNA into the host cell genome is a critical step in the life cycle of HIV. This essential reaction is catalyzed by integrase (IN) through two steps, 3'-processing and DNA strand transfer. Integrase is an attractive target for drug design because there is no known cellular analogue and integration is essential for successful replication of HIV. A computational three-dimensional (3 D) database search was used to identify novel HIV-1 integrase inhibitors. Starting from the previously identified Y3 (4-acetylamino-5-hydroxynaphthalene 2,7-disulfonic acid) binding site on the avian sarcoma virus integrase (ASV IN), a preliminary search of all compounds in the nonproprietary, open part of the National Cancer Institute 3-D database yielded a collection of 3100 compounds. A more rigorous scoring method was used to rescreen the 3100 compounds against both ASV IN and HIV-1 IN. Twenty-two of those compounds were selected for inhibition assays against HIV-1 IN. Thirteen of the 22 showed inhibitory activity against HIV-1 IN at concentrations less than 200 microM and three of them showed antiviral activities in HIV-1 infected CEM cells with effective concentrations (EC50) ranging from 0.8 to 200 microM. Analysis of the computer-generated binding modes of the active compounds to HIV-1 IN showed that simultaneous interaction with the Y3 site and the catalytic site is possible. In addition, interactions between the active compounds and the flexible loop involved in the binding of DNA by IN are indicated to occur. The structural details and the unique binding motif between the HIV-1 IN and its inhibitors identified in the present work may contribute to the future development of IN inhibitors. PMID- 11058034 TI - Non-peptidic, non-prenylic bisubstrate farnesyltransferase inhibitors. Part 3: structural requirements of the central moiety for farnesyltransferase inhibitory activity. AB - Recently, we have described non-peptidic, non-prenylic bisubstrate analogues as a novel type of farnesyltransferase inhibitor composed of a farnesyl-mimetic, a linker and an AAX-peptidomimetic substructure. With this study, we showed that the amide function connecting the farnesyl-mimetic and the linking substructures of our inhibitors is crucial for their activity. We suggest that the amide is bound to the essential zinc ion in the farnesyltransferases active center. We identified succinic and glutaric acid, respectively, in addition to the initially used 1-alanyl moiety as suitable linking structures. Glycine can also be used in this function provided the distance between the alpha-amide group and the center of the peptidomimetic substructure is enlarged by introduction of an additional methylene unit into the peptidomimetic substructure. PMID- 11058035 TI - Synthesis and evaluation of microtubule assembly inhibition and cytotoxicity of prenylated derivatives of cyclo-L-Trp-L-Pro. AB - The synthesis of three isoprenylated derivatives of cyclo-L-Trp-L-Pro is described. These substances have been evaluated for cytotoxic activity in rat normal fibroblast 3Y1 cells and have also been evaluated in vitro for the inhibition of microtubule assembly. PMID- 11058036 TI - Synthesis and biological evaluation of aryl azide derivatives of combretastatin A 4 as molecular probes for tubulin. AB - Two new aryl azides, (Z)-1-(3'-azido-4'-methoxyphenyl)-2-(3",4",5" trimethoxyphenyl)ethene 9 and (Z)-1-(4'-azido-3'-methoxyphenyl)-2-(3",4",5" trimethoxyphenyl)ethene 5, modeled after the potent antitumor, antimitotic agent combretastatin A-4 (CA-4), have been prepared by chemical synthesis as potentially useful photoaffinity labeling reagents for the colchicine site on beta-tubulin. Aryl azide 9, in which the 3'-hydroxyl group of CA-4 is replaced by an azido moiety, demonstrates excellent in vitro cytotoxicity against human cancer cell lines (NCI 60 cell line panel, average GI50 = 4.07 x 10(-8) M) and potent inhibition of tubulin polymerization (IC50 = 1.4+/-0.1 microM). The 4' azido analogue 5 has lower activity (NCI 60 cell line panel, average GI50 = 2.28 x 10(-6) M, and IC50 = 5.2+/-0.2 microM for inhibition of tubulin polymerization), suggesting the importance of the 4'-methoxy moiety for interaction with the colchicine binding site on tubulin. These CA-4 aryl azide analogues also inhibit binding of colchicine to tubulin, as does the parent CA-4, and therefore these compounds are excellent candidates for photoaffinity labeling studies. PMID- 11058037 TI - A practical synthesis of the serotonin 5-HT2A receptor antagonist MDL 100907, its enantiomer and their 3-phenolic derivatives as precursors for [11C]labeled PET ligands. AB - A practical synthesis of the 3-phenolic precursor of MDL 100907, a selective 5 HT2A receptor antagonist, is described. The route was also applied to the enantiomeric series, thus affording the direct precursors of both 3-[11C]MDL 100907 and its enantiomer as ligands for positron emission tomography. Similar methodology was developed for the direct synthesis of MDL 100907 and its enantiomer, MDL 100009. The routes utilized classical optical resolution of the N nor intermediates in at least 98% enantiomeric excess and easily afforded multigram amounts of the chiral precursors of a variety of N- and 3-O-substituted enantiomers. PMID- 11058038 TI - Synthesis of a triply phosphorylated pentapeptide from human tau-protein. AB - Two different strategies for the synthesis of a triply phosphorylated pentapeptide are described. In both cases a monophosphorylated selectively N deprotected tripeptide is employed as C-terminal fragment. Coupling of this building block with a C-terminally unmasked bis-phosphorylated seryl-dipeptide unexpectedly failed due to decomposition of this peptide upon activation with different coupling reagents. Instead stepwise N-terminal elongation of the peptide chain with serine derivatives and subsequent O-phosphorylation of the serine OH-groups was successful. These results indicate that assembly of multiply phosphorylated peptides from preformed multiply phosphorylated phosphopeptide building blocks in general may be problematic and that a stepwise elongation of the amino acid chain may be preferable. PMID- 11058039 TI - Synthesis of novel adamantylacetyl derivative of peptidoglycan monomer- biological evaluation of immunomodulatory peptidoglycan monomer and respective derivatives with lipophilic substituents on amino group. AB - Novel synthetic analogue of immunomodulatory peptidoglycan monomer 1 (PGM), (adamant-1-yl)-CH2CO-PGM (2), was prepared by acylation of epsilon-amino group of diaminopimelic acid with symmetrical (adamant-1-yl)-acetic acid anhydride in the presence of triethylamine. The product was isolated by gel filtration on Sephadex G-25, followed by ion exchange chromatography on SP-Sephadex C-25. The susceptibility of (adamant-1-yl)-CH2CO-PGM to hydrolysis with N-acetylmuramyl-L alanine amidase was demonstrated, and the product of hydrolysis, (adamant-1-yl) CH2CO-pentapeptide 3, was characterized. Both 2 and 3 are water soluble and non pyrogenic compounds. Immunomodulatory activity of PGM (adamant-1-yl)-CH2CO-PGM and structurally related derivative Boc-Tyr-PGM was compared in experiments in vivo, in mice, using ovalbumin (OVA) as an antigen. All three tested compounds exhibited comparable immunostimulating effects with respect to the induction of anti-ovalbumin immunoglobulin G. The results of evaluation of biological activity show that the substitution of free amino group in the parent peptidoglycan molecule with bulky lipophilic substituents did not affect the susceptibility to hydrolysis with N-acetylmuramyl-L-alanine amidase and did not alter markedly the immunostimulating activity. The results also indicate that the free amino group in the peptide chain is not a necessary requirement in the mechanism of immunostimulation of tested immunomodulators. PMID- 11058040 TI - Investigating the role of the geminal dimethyl groups of coenzyme A: synthesis and studies of a didemethyl analogue. AB - An analogue 2 of coenzyme A (CoA) has been prepared in which the geminal methyl groups are replaced with hydrogens. An NMR titration study was conducted and shifts in frequency of protons in the pantetheine portion of the molecule upon titration of the adenine base were observed as has been previously reported with CoA. These studies indicate that the geminal dimethyl groups are not essential for adoption of a partially folded conformation in solution. Based on 1H-1H coupling constants, the distribution of conformations about the carbon-carbon bonds in the region of the methyl deletion were estimated. The results suggest that the conformer distribution is similar to that of CoA, but with small increases in population of the anti conformers. A simple model compound containing the didemethyl pantoamide moiety was prepared and subjected to similar conformational analysis. The coupling constants and predicted conformer distribution were almost identical to that of the CoA analogue, indicating that the conformer distribution is controlled by local interactions and not influenced by interactions between distant parts of the CoA molecule. The acetyl derivative of 2 was a fairly good substrate for the acetyl-CoA utilizing enzymes carnitine acetyltransferase, chloramphenicol acetyltransferase, and citrate synthase, with 1.3- to 10-fold increased Km values and 2.5- to 11-fold decreases in Vmax. The combined results indicate that the geminal dimethyl groups of CoA have modest effects on function and minimal effects on conformation. PMID- 11058041 TI - Synthesis of substituted indeno[1,2-b]quinoline-6-carboxamides, [1]benzothieno[3,2-b]quinoline-4-carboxamides and 10H-quindoline-4-carboxamides: evaluation of structure-activity relationships for cytotoxicity. AB - New substituted indeno[1,2-b]quinoline-6-carboxamides, [1]benzothieno[3,2 b]quinoline-4-carboxamides and 10H-quindoline-4-carboxamides were prepared from methyl 2-amino-3-formylbenzoate by a new Friedlander synthesis. Evaluation of these carboxamides for cytotoxicity in a panel of cell lines showed that small lipophilic substituents in the non-carboxamide ring, in a pseudo-peri position to the side chain, significantly increased cytotoxic potency while retaining a pattern of cytotoxicity consistent with a non-topo II mode of action. The methyl substituted indeno[1,2-b]quinoline-6-carboxamide demonstrated substantial effectiveness (20-day growth delays) in a sub-cutaneous colon 38 in vivo tumor model. This is comparable to that reported for the dual topo I/II inhibitor DACA that is in clinical trial. PMID- 11058042 TI - Alternative heterocycles for DNA recognition: a 3-pyrazole/pyrrole pair specifies for G.C base pairs. AB - Synthetic ligands comprising three aromatic amino acids, pyrrole (Py), imidazole (Im), and hydroxypyrrole (Hp), specifically recognize predetermined sequences as side-by-side pairs in the minor groove of DNA. To expand the repertoire of aromatic rings that may be utilized for minor groove recognition, three five membered heterocyclic rings, 3-pyrazolecarboxylic acid (3-Pz), 4 pyrazolecarboxylic acid (4-Pz), and furan-2-carboxylic acid (Fr), were examined at the N-terminus of eight-ring hairpin polyamide ligands. The DNA binding properties of 3-Pz, 4-Pz, and Fr each paired with Py were studied by quantitative DNase I footprinting titrations on a 283 bp DNA restriction fragment containing four 6-bp binding sites 5'-ATNCCTAA-3' (N = G, C, A, or T; 6-bp polyamide binding site is underlined). The pair 3-Pz/Py has increased binding affinity and sequence specificity for G.C bp compared with Im/Py. PMID- 11058043 TI - Design and synthesis of potential inhibitors of the ergosterol biosynthesis as antifungal agents. AB - A series of azolylmethyloxolane derivatives with modified sterol side-chain structures, designed as potential dual functional inhibitors of cytochrome P450 14alpha-demethylase (14DM) and delta24-sterol methyltransferase (24-SMT) based on the common characteristic features of 24-aminosterols and azole antifungal agents, were synthesized and evaluated for their antifungal activities and inhibitory activities of 14DM and 24-SMT. Among these compounds, imidazolylmethyloxolane derivatives 28a and 28b showed potent in vitro antifungal activities comparable to those of itraconazole. However, the in vitro bioactivities have not been linearly translated into in vivo protection data for some unknown reasons. PMID- 11058044 TI - Understanding the antifungal activity of terbinafine analogues using quantitative structure-activity relationship (QSAR) models. AB - Terbinafine and its analogues, which are a major class of non-azole antifungal agents, are known to act by inhibition of squalene epoxidase enzyme in fungal cells. We have performed a quantitative structure-activity relationship (QSAR) study on a series of 92 molecules using different types of physicochemical descriptors. Inhibitors were divided into five classes depending upon chemical structure. QSAR models were generated for correlation between antifungal activity against Candida albicans using genetic function approximation (GFA) technique. Equations were evaluated using internal as well as external test set predictions. Models generated for all these classes show that steric properties and conformational rigidity of side chains play an important role for the activity. The present QSAR analysis agrees with the results of the previously reported CoMFA study. PMID- 11058045 TI - Synthesis of phosphonate derivatives of uridine, cytidine, and cytosine arabinoside. AB - The vinyl phosphonate derivatives of uridine, cytidine, and cytosine arabinoside (ara-C) have been prepared through oxidation of appropriately protected nucleosides to the 5' aldehydes and Wittig condensation with [(diethoxyphosphinyl)methylidine]triphenylphosphorane. Dihydroxylation of these vinyl phosphonates with an AD-mix reagent generated the new 5',6'-dihydroxy-6' phosphonates. After hydrolysis of the phosphonate esters and the various protecting groups, the six phosphonic acids were tested for their ability to serve as substrates for the enzyme nucleotide monophosphate kinase and for their toxicity to K562 cells. PMID- 11058046 TI - Radiosynthesis of [18F]Lu29-024: a potential PET ligand for brain imaging of the serotonergic 5-HT2 receptor. AB - In a previous work, Lu29-024 (2,5-dimethyl-3-(4-fluorophenyl)-1-(1-methyl-4 piperidinyl)-1H-indole), a selective 5-HT2A receptor antagonist with nanomolar affinity and high selectivity, was labeled with carbon-11 to evaluate its behavior as a potential PET ligand for the serotonergic 5-HT2A receptor in the central nervous system. Administration of this tracer to rats was followed by a good brain uptake, no brain labeled metabolites but no specific, regio-selective, binding at 20 and 40 min post injection. Despite this, the data noted at 20 and 40 min suggest that this tracer, if associated with a radioactive emitter with a longer half-life than that of carbon-11, could be useful for the quantification of 5HT2A receptors. For these reasons, we chose to label this compound, bearing a fluorine atom, with [18F]fluoride, in order to perform rat studies over a more prolonged time-scale. The precursor for the radiosynthesis of [18F]Lu29-024 was obtained in an overall yield of 20% by a multi-step synthesis including an acetonylation reaction followed by a Fisher indole reaction. The radiotracer was prepared by an aromatic substitution with activated [18F]fluoride followed by a decarbonylation reaction that employed Wilkinson's catalyst. The radiosynthesis of [18F]Lu29-024 required approximatively 110 min with an overall radiochemical yield of 20-35% and specific activities of 37GBq/micromol. Fluorine-labeled Lu29 024 may thus be envisaged as a potentially useful PET tracer that can be applied to a wide range of neurological and psychiatric diseases. PMID- 11058047 TI - Chemoenzymatic synthesis of L-galactosylated dimeric sialyl Lewis X structures employing alpha-1,3-fucosyltransferase V. AB - L-Galactosylated dimeric sialyl Lewis X (SLeX) has been prepared employing a combination of chemical and enzymatic synthetic methods. GDP-L-galactose has been chemically synthesised. Enzymatic transfer of L-galactose onto the acceptor (Sia alpha2,3-Gal-beta1,4-GlcNAc-beta1,3/6)2-Man-alpha1-OMe was achieved using the human alpha-1,3-fucosyltransferase V. PMID- 11058048 TI - Photophysical properties of corrphycenes. AB - The photophysical properties of the Zn salt of octaethylcorrphycene (compound 1) and the doubly protonated octaethylcorrphycene (compound 2) were determined in benzene solutions. Fluorescence spectra and fluorescence quantum yields of phiF (1) = 0.03+/-0.02 and PF (2) = 0.06+/-0.02 were measured. The triplet-triplet absorption spectra were obtained by means of flash-photolysis experiments. The triplet quantum yield values phiT (1) = 0.79+/-0.08 and phiT (2) = 0.82+/-0.08 were obtained by using laser-induced optoacoustic spectroscopy. The quantum yield of singlet molecular oxygen generation in air-saturated solutions, phidelta (1) = 0.55+/-0.07 and phidelta (2) = 0.38+/-0.05, were also measured using time resolved NIR luminescence. PMID- 11058049 TI - Electronic and vibrational spectra of a series of substituted carbazole derivatives. AB - The FTIR and FTR spectra of halogen (Cl, Br, I) substituted carbazole and their N acetic and propionic acids have been recorded. A number of lines have been assigned on the basis of previous studies on the parent compound and by comparisons with the characteristic vibrations of their constituent structural units as well as comparing the spectra from FTIR and FTR. Some substituent sensitive bands and characteristic bands were found. The electronic absorption spectra of these compounds in acetonitrile were also measured and are briefly discussed. PMID- 11058050 TI - g factors and the tetragonal distortion of ligand octahedron for Co2 + in Rb2MgF4 crystal. AB - The calculation formulas of g-factors g(parallel) and g(perpendicular) for 3d7 ion in tetragonal octahedral crystals are established from a cluster approach. In these formulas, the parameters related to covalency effect, configuration interaction and low-symmetry crystal field can be determined from the optical spectra and the structural data of the studied system. Based on these formulas, the structural parameters of ligand octahedra of Co2+ in Rb2MgF4 crystal are obtained by fitting the calculated g(parallel) and g(perpendicular) to the observed values. The result suggests that the CoF6 (and hence MgF6) octahedra in Rb2MgF4:Co2+ are tetragonal compressed. The relationship between the sign of deltag( = g(perpendicular) - g(parallel)) and the sign of distortion (elongated or compressed) of ligand octahedron and the causes of the mistakes of octahedron distortion for Rb2MgF4:Co2+ in the previous papers are discussed. PMID- 11058051 TI - Surface-enhanced raman spectra of dyes and organic acids in silver solutions: chloride ion effect. AB - We have recorded surface-enhanced Raman (SER) spectra of two different classes of compounds, cationic dyes and organic acids, and studied their chloride ion effects on the surface-enhanced Raman scattering (SERS) activities of the silver solution. For the positive charge dyes, rhodamine 6G (R6G) and 1,1'-dimethyl-2,2' cyanine iodide (DECI), no SERS could be observed without the addition of chloride ions because of lack of the electrostatic interaction between the dye species and the silver particles in the silver solution. The chloride ions served to enlarge silver particles and to contribute the existence of the surface active sites, making the silver solution SERS active to the dye samples. Surface-enhanced resonance Raman scattering (SERRS) intensity of the dye molecules increased with the chloride ion concentration. After reaching a maximum intensity, a Cl- quenching effect on the intensity took place. For the organic acids, benzoic acid and p-aminobenzoic acid (PABA), SERS could be observed without the coexistence of chloride ions. The intensity of the Raman scattering did not vary significantly in the presence of small amount of chloride ion. At high Cl- concentration, quenching SERS intensity began to take effect. PMID- 11058052 TI - Photoacoustic spectroscopy study on intramolecular energy transfer and relaxation processes of Tb(III) complexes. AB - The photoacoustic (PA) amplitude spectra and luminescence spectra of different Tb(III) complexes (Tb(AA)3.2H2O Na[Tb(AA)4], Tb(AA)3bpy and Tb(AA)3phen) have been measured, and the PA phase shifts of the different complexes calculated. Combined with the luminescence spectra, the PA amplitude spectra reflected the variation of the luminescence efficiency and the PA phase is directly relative to the relaxation processes. According to the variation of the luminescence efficiency and the phase shift, the intramolecular energy transfer and relaxation processes of different Tb(III) complexes were discussed. PMID- 11058053 TI - Estimation of the total range of 1J(CC) couplings in heterocyclic compounds: pyridines and their N-oxides, the experimental and DFT studies. AB - A very large set of one-bond spin-spin carbon carbon coupling constants, 1J(CC), has been measured for 32 variously mono- and disubstituted pyridine N-oxides and for 14 substituted pyridines. The N-oxides studied were 2-, 3- and 4 monosubstituted isomers, and a series of disubstituted compounds. A variety of substituents has been employed (CH3, COCH3, C5H4NO, CN, F, Br, Cl, OH, OCH3, NH2, N(CH3)2 and NO2), which allowed us to study substituent effects thoroughly. Good linear relationships between 1J(C3C4) in 3- and/or 4-substituted pyridine N oxides and 1J(CipsoCortho) in benzenes and between 1J(C2C3) in 2- and/or 3 substituted pyridine N-oxides and 1J(CipsoCortho) in benzenes have been found. An analogous linear relationship has been observed between 1J(C3C4) in 3- and/or 4 substituted pyridines and 1J(CipsoCortho) in benzenes. It has been also concluded that, by analogy to 1J(CC) couplings in substituted benzenes, those in pyridines and their N-oxides are the substituent electronegativity dependent. The estimated total range covered by 1J(CC), couplings in substituted compounds varies, in the case of 1J(C2C3) couplings for example, from 25 Hz in 2-lithiopyridine N-oxide to ca. 100 Hz in 2,3-difluoropyridine N-oxide and from 18 Hz in 2-lithiopyridine to 92 Hz in 2,3-difluoropyridine. The DFT calculations have been carried out for the parent compounds and for a set of their 2-lithio, and variously substituted fluoro derivatives. The DFT data reproduced very well the experimental coupling values and revealed that the Fermi contact contribution is the dominating factor which governs the magnitude of the CC coupling across one bond. PMID- 11058054 TI - Infrared and Raman spectra, conformational stability, ab initio calculations and vibrational assignments for trans-3-chloropropenoyl chloride. AB - The infrared (3500-30 cm(-1)) spectra of gaseous and solid and the Raman (3500 200 cm(-1)) spectra of the liquid with quantitative depolarization ratios and solid trans-3-chloropropenoyl chloride (trans-ClCHCHCClO) have been recorded. These data indicate that both the anti (carbonyl bond trans to the carbon-carbon double bond) and syn conformers are present in the fluid states but only the anti conformer is present in the crystalline state. The mid-infrared spectra of the sample dissolved in liquid xenon as a function of temperature (-55 to -100 degrees C) have been recorded. Utilizing conformer pairs at 870 and 725 cm(-1), 1215 and 1029 cm(-1), and 1215 and 1228 cm(-1), the enthalpy difference has been determined to be 136+/-5 cm(-1) (389+/-14 cal mol(-1)) with the anti conformer the more stable form. Optimized geometries and conformational stabilities were obtained from ab initio calculations at the levels of RHF/6-31G(d), MP2/6-31G(d), MP2/6-311 + + G(d,p), MP2/6-311 + + G(2d,2p) and MP2/6-311 + + G(2df,2pd) with only the latter two calculations predicting the anti rotamer to be the more stable form. The vibrational frequencies, harmonic force constants and infrared intensities were obtained from the MP2/6-31G(d) calculations, whereas the Raman activities and depolarization values were obtained from the RHF/6-31G(d) calculations. The spectra are interpreted in detail and the results are compared with those obtained for some related molecules. PMID- 11058055 TI - Surface enhanced Raman scattering of 2,2' biquinoline adsorbed on colloidal silver particles. AB - Surface enhanced Raman scattering (SERS) in silver sol and normal Raman spectra in the bulk and in solution of 2,2' biquinoline (BQ) molecule have been investigated. The observed Raman bands along with their corresponding FTIR bands have been assigned based on the established assignments of the vibrational bands of the parent napthalene and quinoline molecules. Existence of both the cis and trans form of the BQ molecule in solution and in the bulk are inferred from the normal Raman and FTIR spectra, whereas SERS study reveal that in the surface adsorbed state the molecule exists in the cis form. Definite evidence of the charge transfer interaction to the overall contribution in the SER enhancement have been reported. The excitation profile also supports the CT interaction. Estimated enhancement factor of the principal SERS bands indicate that the molecule is adsorbed on the silver surface through both the nitrogen atoms with the molecular plane almost perpendicular to the surface. This preferred orientation of the molecule is in conformity with its existence in the cis form in the surface adsorbed state. PMID- 11058056 TI - ESR and resonance Raman spectroscopic studies of peroxide intermediates derived from iron diazaoxacyclononane. AB - A peroxide-Fe3+ intermediate generation during the Fenton reaction of iron chelate involving a ligating N,N'-di-2-picolyl-4, 7-diaza-1-oxacyclononane (DPC), H2O2/[Fe2+ DPC]2+, is reported. The identity of this peroxide complex is confirmed by resonance Raman (RR) and electron spin resonance (ESR) spectroscopies. The RR spectrum of [Fe2+ DPC]2+ treated with H2O2 shows a frequency at 854 cm(-1) ascribable to v(O-O) vibrational modes of the peroxide Fe3+ complex with a side-on geometry. On the other hand, the ESR spectrum of H2O2/[Fe2+ DPC]2+ acquired at 77 K exhibits the resonance transition at g = 2.196 and 2.017 due to the peroxide-Fe3+ complex with an axial symmetry. It has been concluded that the H2O2/[Fe2+ DPC]2+ reaction proceeds by rapid bonding of H2O2 to an open coordination site on the central Fe2+ cation. PMID- 11058057 TI - A tunable single-mode 3.2 microm laser based on an InAsSb/InAsSbP double heterostructure with drive-current tuning range of 10 cm(-1). AB - A new type of semi-conductor laser with composition InAsSb/InAsSbP is described. This laser was produced for the absorption spectroscopy of atmospherically important molecules in the 3100 cm(-1) region and tested using a closed-cycle He cryostat in the temperature range 30-80 K. The optimal characteristics of the laser were found to be a heatsink temperature of 62 K and a drive current range of 50-350 mA. Under these conditions, the laser emits single-mode radiation in an exceptionally large wavenumber range of > 10 cm(-1). To test the laser, several experiments were carried out in which the rovibrational absorption spectra of CH3Cl, NH3, OCS and H2O were measured. PMID- 11058058 TI - Vibrational spectroscopic studies on ion solvation of lithium perchlorate in propylene carbonate + N,N-dimethylformamide mixtures. AB - The infrared (IR) and Raman spectra are reported for solutions of lithium perchlorate in propylene carbonate (PC), N,N-dimethylformamide (DMF) and PC + DMF mixtures. The band splittings of symmetric ring deformation for PC and O=CN deformation for DMF suggest that there is a strong interaction between lithium cations and solvent molecules. The solvent molecules have been assigned to two types, the free and complexed molecules. By a comparison of the intensity for the corresponding bands, it has been concluded that Li+ cations are preferentially solvated by DMF molecules in the LiClO4/PC-DMF solutions. This has been explained by the difference in values of donor number. PMID- 11058059 TI - Ab initio inter-radical potential for p-phenylenediamine radical cation dimer. AB - Ab initio molecular orbital calculations were carried out to investigate the inter-radical interaction of the paired p-phenylenediamine radical cations in the singlet state. After initial optimization of the dimer in the parallel sandwich (D2h) and parallel displaced (Cs) configurations at the B3LYP/6-31G* theoretical level, the MP2/6-31G* and B3LYP/6-31G* single energies of the dimer were calculated as a function of the inter-radical distance R. The depths of the potential minima near R = 3.2 A were estimated to be in the order of the hydrogen bonding energy, assuming that the electrostatic contribution between the cations is canceled out by the attractive contributions due to the counter anions on the aspect of a simple electrostatic model. This can be related to the indications of the cation dimer formation in solution in the presence of counter anions at a low temperature reported previously in the literature by resonance Raman and electronic absorption spectra. The inter-radical (Raman active) frequencies of the dimer were calculated, one of which corresponds to the reported value at 161 cm(-1) observed in the resonance Raman spectrum in ethanol at 200 K by Yokoyama and Maeda (Chem. Phys. Lett. 48 (1977) 59). PMID- 11058060 TI - Self-association of pyridine-2,6-dicarboxylic acid in aqueous solution as determined from ultraviolet hypochromic and hyperchromic effects. AB - The self-association of pyridine-2,6-dicarboxylic acid (dipicolinic acid), was studied in aqueous solution at the pH values of 0.2, 3.5, 6.9 and 13.0, by ultraviolet spectroscopy. The variation in molar absorptivity with the concentration of this compound was measured for the band in the mid-ultraviolet region. Deviations from Beer-Lambert law with increasing concentration were found at the pH values of 0.2, 3.5 and 6.9. Hypochromic effects were detected at pH 0.2 and 6.9, while a hyperchromic effect was observed at pH 3.5. These results were interpreted in terms of self-association leading to the formation of dimers. From the fitting of the experimental curves of hypochromic or hyperchromic effects, self-association constants were calculated. The occurrence of hypochromic and hyperchromic effects was analyzed in terms of vertical and horizontal interactions, respectively, to elucidate the nature of the self-association processes, stacking or hydrogen bonding, relevant to each of the protolytic species of dipicolinic acid in aqueous solution. PMID- 11058061 TI - The dependence of the fluorescence properties, laser properties and photochemical stability of aromatic compounds on the molecular symmetry. AB - It is known that the spectral properties of organic compounds are, to a large extent, determined by the molecular symmetry. Numerous articles and monographs have been devoted to this problem. However, the influence of the molecular symmetry on fluorescence and, hence, laser parameters has not been fully investigated. In this paper, the fluorescence and laser properties of 20 aromatic compounds are experimentally studied at room temperature. The compounds studied are arranged in family-related pi-structure pairs. In each pair, even-numbered compounds belong to a higher symmetry group than odd-numbered compounds due to symmetrical substitution. All main fluorescence parameters such as quantum yield, gamma, decay time, tauf, fluorescence rate constant, k(f) (Einstein coefficient, A), and intersystem crossing rate constant, kST, are measured or calculated. It has been found that for most of the symmetrically substituted molecules, the value of kST decreases, sometimes very significantly. For example, the transition from 9-phenylanthracene (C2 symmetry) to 9,10-diphenylanthracene (D2 symmetry) is accompanied by an 18-fold decrease in the value of kST. This phenomenon is explained by the fact that, in a molecule of higher symmetry, not all triplet states mix with the fluorescing S1 state. It is also found that the symmetry of a molecule greatly affects laser parameters such as the threshold of laser action and the photochemical stability of a laser solution. It is observed that the threshold for even-numbered compounds is much lower and the photochemical stability, in most cases, is much higher than for odd-numbered compounds. These phenomena are discussed and explained. PMID- 11058062 TI - A multinuclear NMR study of [Pt0(PPh3)2(alkene)] compounds containing asymmetric olefins. AB - The 1H, 13C, 31P, and 195Pt NMR spectra of [Pt0(PPh3)2(eta-ABC(1) = C(2)XY)] compounds (ABC(1)= C(2)XY (1) A = B = X = Y = H; (3) A = B = X = H, Y = CN; (4) A = H, B = p-NO2-Ph, X = COOCH3, Y = CN; (5) A = H, B = Ph, X = COOCH3, Y = CN; (6) A = H, B = Ph, X = Y = CN; (7) A = H, B = OEt, X = Y = CN), where X and Y are electronacceptor substituents, and the 1H spectrum of [Pt0(PPh3)2(eta2-C60)] (2) are reported together with extended analyses and assignments, based also on the ring current effect of the olefin phenyl in (4-6). Deviations from first order in the 13C spectra allowed the determination of the relative signs of the coupling constants J(P(1), C) and J(P(2), C) of the alkene and of the triphenylphosphine carbons. Best fit simulation of the phosphine C ipso spectrum provided also the 13C isotopic shift on phosphorus for (1). These compounds are characterised by strong differences between the two platinum-phosphorus coupling constants in the case of asymmetric olefins (3-7). The chemical shifts of alkene C(1) and C(2) indicate notable polarisation of the olefin after complexation, while the 1J(Pt, C(1)) and 1J(Pt, C(2)) values are in agreement with a stronger interaction of Pt with C(1) than with C(2). These findings together with the trend of 195Pt chemical shifts confirm the important role played by back-donation in the bonding of platinum(0)-olefin compounds. PMID- 11058063 TI - Vibrational spectroscopy of ferruginous smectite and nontronite. AB - A comparison is made between the Raman and infrared spectra of ferruginous smectite and a nontronite using both absorption and emission techniques. Raman spectra show hydroxyl stretching bands at 3572, 3434, 3362, 3220 and 3102 cm(-1). The infrared emission spectra of the hydroxyl stretching region are significantly different to the absorption spectrum. These differences are attributed to the loss of water, absent in the emission spectrum, the reduction of the samples in the spectrometer and possible phase changes. Dehydroxylation of the two minerals may be followed by the loss of intensity of the hydroxyl stretching and hydroxyl deformation frequencies. Hydroxyl deformation modes are observed at 873 and 801 cm(-1) for the ferruginous smectite, and at 776 and 792 cm(-1) for the nontronite. Raman hydroxyl deformation vibrations are found at 879 cm(-1). Other Raman bands are observed at 1092 and 1032 cm(-1), assigned to the SiO stretching vibrations, at 675 and 587 cm(-1), assigned to the hydroxyl translation vibrations, at 487 and 450 cm(-1), attributed to OSiO bending type vibrations, and at 363, 287 and 239 cm(-1). The differences in the molecular structure of the two minerals are attributed to the Al/Fe ratio in the minerals. PMID- 11058064 TI - 13C and 1H NMR of 2,6-diaryl-1-hydroxy piperidin-4-one oximes; substituent effects on cis/trans ratio and conformational equilibria. AB - The reaction of substituted diarylidene acetones with hydroxylamine hydrochloride affords isomeric N-hydroxy diaryl piperidinone oximes as main products. The structures as well as conformational equilibria of these products were established by 1H and 13C NMR spectroscopy and further studied by variable temperature NMR. It was found that the cis/trans ratio of 2,6-substituted piperidine derivatives depended on the position of the substituent on the aromatic ring. PMID- 11058065 TI - Spectroscopic and structural studies of the complex of [Cu6(bpy)10(mu-CO3)2(mu OH)2](ClO4)6 . 4H2O. AB - The electronic absorption spectrum (diffuse reflection spectrum) of the crystal of [Cu6(bpy)10([mu-CO3)2(mu-OH)2](ClO4)6 . 4H2O has been measured. The experimental results are discussed quantitatively with ligand field theory and the radial wave function of non-free copper(II), and calculation values agree well with the experimental results. The d-d absorption spectrum of a novel hexanuclear copper(II) complex was explained satisfactorily. Especially, complexity of multinuclear crystal structures determined that of spectral behaviors. It provides significant to grope spectral nature from coordination structures. PMID- 11058066 TI - A re-examination of the line shape of the electronic spectra of complex molecules in solution: log-normal function versus gaussian. AB - The log-normal function and the Gauss function have been tested for the description of highly structured UV/Vis spectra of complex molecules in solution. The Gauss function was shown to give the better fit for these spectra and is also useful for fluorescence spectra. These results were further supported by the application of the Ross function. PMID- 11058067 TI - Far-infrared spectroscopy of CH3OD in highly excited torsional states and the atlas of the Fourier transform spectra in the range 200-350 cm(-1). AB - The high resolution Fourier transform far-infrared (FIR) spectrum of the torsion rotation band of CH3OD has been analyzed for the highly excited torsion states (n > or = 2) in the vibrational ground state. The spectrum shows splitting of the lines due to strong torsional-rotational-vibrational interactions in the molecule. Assignments were possible for rotational sub-bands in the torsional state as high as n = 4 and for K values up to 8 and J values of up to approximately 30 in most cases, for all the symmetry species. For the third excited torsional state n = 3 assignments were possible to K = 10. The data were analyzed with the help of the energy expansion model, which has been proven very successful in methanol. The state dependent expansion parameters are presented. These molecular parameters were able to reproduce the observed wavenumbers almost to within experimental accuracy of 0.0002 cm(-1) for clean unblended lines. These expansion coefficients should prove valuable in the calculation of precise energy values for excited torsional states up to n = 4, which is way above the torsional barrier. The detailed high-resolution spectral atlas of CH3OD has been presented in the range 200-350 cm(-1). This atlas is an extension of our earlier atlas in the range 20-205 cm(-1). The availability of this atlas in the journal will be very valuable for spectroscopists and astrophysicists seeking information in the infrared (IR) region in the laboratory and in outer space. PMID- 11058068 TI - Green up-conversion emission in Er3+:BaTiO3 sol-gel powders. AB - Bright green emissions at 549 and 526 nm have been observed from sol-gel derived Er3+:BaTiO3 powders upon excitations at two near infrared (NIR) wavelengths of 973 and 816 nm. The decay characteristics were measured and studied. It was found that the predominant emission at 549 nm has very different lifetimes upon excitation at 973 and 816 nm, which was explained by the mechanisms of excited state absorption (ESA) of individual Er3+ ion and cooperative energy transfer (CET) between two near Er3+ ions for the up-conversion emission. Analysis also showed that 973-nm excitation is more effective for the green up-conversion emission. PMID- 11058069 TI - Effect of leucinyl-phenylalanyl-valine on DMPC liposome membrane. AB - Leucinyl-phenylalanyl-valine (LFV) is a hydrophobic tripeptide with a flat egg shaped structure with the long axis dimension of about 12 A. The effect of LFV on dimyristoylphosphatidylcholine (DMPC) liposome membrane has been studied by differential scanning calorimetry (DSC) and fluorescence spectroscopy. Calorimetric studies shows that incorporation of LFV completely abolishes the pretransition temperature with broadening of main transition temperature. Four conceptually different fluorescence probes, 1-naphthol (1-ROH) an excited state proton transfer probe, 8-anilino-1-naphthalenesulphonate (ANS) a solvent polarity probe, 1-6-diphenylhexatriene (DPH) an anisotropy probe and pyrene an excimer forming probe have been used for fluorescence spectroscopic studies. For 1-ROH, ANS and DPH, a decreased partitioning with increasing mol.% of LFV was observed. Increasing LFV mol.% caused a decrease in the neutral form emission of 1-ROH, and a decrease in fluorescence intensity with red shift in ANS. The excimer formation ability of pyrene also decreased. The phase transition behavior of DMPC membrane in the presence of LFV was similar to the known effect of cholesterol on lipid bilayers. These results suggest that LFV cause an increased compactness of membrane. PMID- 11058070 TI - 1-Naphthol as an excited state proton transfer fluorescent probe for erythrocyte membranes. AB - The microenvironmental dependence of excited state prototropism of 1-naphthol and the corresponding changes in its fluorescence emission is utilized to monitor the acyl chain melting phase transition behavior of liposome membrane made from human erythrocyte lipids. A sharp increase in the ratio of neutral/anionic form fluorescence intensity is noticed at the phase transition temperature (19 degrees C). This provides a convenient method for obtaining phase transition temperature in lipid membranes. The membrane modifying effect of cholesterol on the erythrocyte liposome is successfully sensed by 1-naphthol fluorescence. PMID- 11058072 TI - The slow death of scientific meetings. By Caveman. PMID- 11058071 TI - Spectroscopic studies on the interaction of bovine (BSA) and human (HSA) serum albumins with ionic surfactants. AB - Bovine (BSA) and human (HSA) serum albumins are frequently used in biophysical and biochemical studies since they have a similar folding, a well known primary structure, and they have been associated with the binding of many different categories of small molecules. One important difference of BSA and HSA is the fact that bovine albumin has two tryptophan residues while human albumin has a unique tryptophan. In this work results are presented for the interaction of BSA and HSA with several ionic surfactants, namely, anionic sodium dodecyl sulfate (SDS), cationic cethyltrimethylammonium chloride (CTAC) and zwitterionic N hexadecyl-N,N-dimethyl-3-ammonium-1-propanesulfonate (HPS), as monitored by fluorescence spectroscopy of intrinsic tryptophans and circular dichroism spectroscopy. On the interaction of all three surfactants with BSA, at low concentrations, a quenching of fluorescence takes place and Stern-Volmer analysis allowed to estimate their 'effective' association constants to the protein: for SDS, CTAC and HPS at pH 7.0 these constants are, respectively, (1.4+/-0.1) x 10(5) M(-1), (8.9+/-0.1) x 10(3) M(-1) and (1.4+/-0.1) x 10(4) M(-1). A blue shift of maximum emission is observed from 345 to 330 nm upon surfactant binding. Analysis of fluorescence emission spectra allowed to separate three species in solution which were associated to native protein, a surfactant protein complex and partially denatured protein. The binding at low surfactant concentrations follows a Hill plot model displaying positive cooperativity and a number of surfactant binding sites very close to the number of cationic or anionic residues present in the protein. Circular dichroism data corroborated the partial loss of secondary structure upon surfactant addition showing the high stability of serum albumin. The interaction of the surfactants with HSA showed an enhancement of fluorescence at low concentrations, opposite to the effect on BSA, consistent with the existence of a unique buried tryptophan residue in this protein with considerable static quenching in the native state. The effects of surfactants at low concentrations were very similar to those of myristic acid suggesting a non specific binding through hydrophobic interaction modulated by eletrostatic interactions. The changes in the vicinity of the tryptophan residues are discussed based on the recently published crystallographic structure of HSA myristate complex (S. Curry et al., Nat. Struct. Biol. 5 (1998) 827). PMID- 11058073 TI - The nuclear pore complex. PMID- 11058076 TI - Chk1 and Cds1: linchpins of the DNA damage and replication checkpoint pathways. AB - Recent work on the mechanisms of DNA damage and replication cell cycle checkpoints has revealed great similarity between the checkpoint pathways of organisms as diverse as yeasts, flies and humans. However, there are differences in the ways these organisms regulate their cell cycles. To connect the conserved checkpoint pathways with various cell cycle targets requires an adaptable link that can target different cell cycle components in different organisms. The Chk1 and Cds1 protein kinases, downstream effectors in the checkpoint pathways, seem to play just such roles. Perhaps more surprisingly, the two kinases not only have different targets in different organisms but also seem to respond to different signals in different organisms. So, whereas in fission yeast Chk1 is required for the DNA damage checkpoint and Cds1 is specifically involved in the replication checkpoint, their roles seem to be shuffled in metazoans. PMID- 11058077 TI - ID helix-loop-helix proteins in cell growth, differentiation and tumorigenesis. AB - The ubiquitously expressed family of ID helix-loop-helix (HLH) proteins function as dominant negative regulators of basic HLH (bHLH) transcriptional regulators that drive cell lineage commitment and differentiation in metazoa. Recent data from cell line and in vivo studies have implicated the functions of ID proteins in other cellular processes besides negative regulation of cell differentiation. ID proteins play key roles in the regulation of lineage commitment, cell fate decisions and in the timing of differentiation during neurogenesis, lymphopoiesis and neovascularisation (angiogenesis). They are essential for embryogenesis and for cell cycle progression, and they function as positive regulators of cell proliferation. ID proteins also possess pro-apoptotic properties in a variety of cell types and function as cooperating or dominant oncoproteins in immortalisation of rodent and human cells and in tumour induction in Id transgenic mice. In several human tumour types, the expression of ID proteins is deregulated, and loss- and gain-of-function studies implicate ID functions in the regulation of tumour growth, vascularisation, invasiveness and metastasis. More recent biochemical studies have also revealed an emerging 'molecular promiscuity' of mammalian ID proteins: they directly interact with and modulate the activities of several other families of transcriptional regulator, besides bHLH proteins. PMID- 11058078 TI - Detyrosinated (Glu) microtubules are stabilized by an ATP-sensitive plus-end cap. AB - Many cell types contain a subset of long-lived, 'stable' microtubules that differ from dynamic microtubules in that they are enriched in post-translationally detyrosinated tubulin (Glu-tubulin). Elevated Glu tubulin does not stabilize the microtubules and the mechanism for the stability of Glu microtubules is not known. We used detergent-extracted cell models to investigate the nature of Glu microtubule stability. In these cell models, Glu microtubules did not incorporate exogenously added tubulin subunits on their distal ends, while >70% of the bulk microtubules did. Ca(2+)-generated fragments of Glu microtubules incorporated tubulin, showing that Glu microtubule ends are capped. Consistent with this, Glu microtubules in cell models were resistant to dilution-induced breakdown. Known microtubule end-associated proteins (EB1, APC, p150(Glued) and vinculin focal adhesions) were not localized on Glu microtubule ends. ATP, but not nonhydrolyzable analogues, induced depolymerization of Glu microtubules in cell models. Timelapse and photobleaching studies showed that ATP triggered subunit loss from the plus end. ATP breakdown of Glu microtubules was inhibited by AMP PNP and vanadate, but not by kinase or other inhibitors. Additional experiments showed that conventional kinesin or kif3 were not involved in Glu microtubule capping. We conclude that Glu microtubules are stabilized by a plus-end cap that includes an ATPase with properties similar to kinesins. PMID- 11058079 TI - Active protein transport through plastid tubules: velocity quantified by fluorescence correlation spectroscopy. AB - Dynamic tubular projections emanate from plastids in certain cells of vascular plants and are especially prevalent in non-photosynthetic cells. Tubules sometimes connect two or more different plastids and can extend over long distances within a cell, observations that suggest that the tubules may function in distribution of molecules within, to and from plastids. In a new application of two-photon excitation (2PE) fluorescence correlation spectroscopy (FCS), we separated diffusion of fluorescent molecules from active transport in vivo. We quantified the velocities of diffusion versus active transport of green fluorescent protein (GFP) within plastid tubules and in the cytosol in vivo. GFP moves by 3-dimensional (3-D) diffusion both in the cytosol and plastid tubules, but diffusion in tubules is about 50 times and 100 times slower than in the cytosol and an aqueous solution, respectively. Unexpectedly larger GFP units within plastid tubules exhibited active transport with a velocity of about 0.12 microm/second. Active transport might play an important role in the long-distance distribution of large numbers of molecules within the highly viscous stroma of plastid tubules. PMID- 11058080 TI - Intact Ca(2+)-binding sites are required for targeting of annexin 1 to endosomal membranes in living HeLa cells. AB - Annexin 1 is a Ca(2+)-regulated membrane binding protein and a major substrate of the epidermal growth factor receptor kinase. Because of its properties and intracellular distribution, the protein has been implicated in endocytic trafficking of the receptor, in particular in receptor sorting occurring in multivesicular endosomes. Up to now, however, the localization of annexin 1 to cellular membranes has been limited to subcellular fractionation and immunocytochemical analyses of fixed cells. To establish its localization in live cells, we followed the intracellular fate of annexin 1 molecules fused to the Green Fluorescent Protein (GFP). We show that annexin 1-GFP associates with distinct, transferrin receptor-positive membrane structures in living HeLa cells. A GFP chimera containing the Ca(2+)/phospholipid-binding protein core of annexin 1 also shows a punctate intracellular distribution, although the structures labeled here do not resemble early but, at least in part, late endosomes. In contrast, the cores of annexins 2 and 4 fused to GFP exhibit a cytoplasmic or a different punctate distribution, respectively, indicating that the highly homologous annexin core domains carry distinct membrane specificities within live cells. By inactivating the three high-affinity Ca(2+) binding sites in annexin 1 we also show that endosomal membrane binding of the protein in live HeLa cells depends on the integrity of these Ca(2+) binding sites. More detailed analysis identifies a single Ca(2+) site in the second annexin repeat that is crucially involved in establishing the membrane association. These results reveal for the first time that intracellular membrane binding of an annexin in living cells requires Ca(2+) and is mediated in part through an annexin core domain that is capable of establishing specific interactions. PMID- 11058081 TI - Fast transport of neurofilament protein along microtubules in squid axoplasm. AB - Using squid axoplasm as a model system, we have visualized the fast transport of non-filamentous neurofilament protein particles along axonal microtubules. This transport occurs at speeds of 0.5-1.0 microm/second and the majority of neurofilament particles stain with kinesin antibody. These observations demonstrate, for the first time, that fast (0.5-1.0 microm/second) transport of neurofilament proteins occurs along microtubules. In addition, our studies suggest that neurofilament protein can be transported as non-membrane bound, nonfilamentous subunits along axons, and that the transport is kinesin-dependent. Microtubule-based fast transport might therefore provide a mechanism for the distribution and turnover of neurofilament, and perhaps other cytoskeletal proteins, throughout neurons. PMID- 11058082 TI - Calsequestrin, a calcium sequestering protein localized at the sarcoplasmic reticulum, is not essential for body-wall muscle function in Caenorhabditis elegans. AB - Calsequestrin is the major calcium-binding protein of cardiac and skeletal muscles whose function is to sequester Ca(2+ )in the lumen of the sarcoplasmic reticulum (SR). Here we describe the identification and functional characterization of a C. elegans calsequestrin gene (csq-1). CSQ-1 shows moderate similarity (50% similarity, 30% identity) to rabbit skeletal calsequestrin. Unlike mammals, which have two different genes encoding cardiac and fast-twitch skeletal muscle isoforms, csq-1 is the only calsequestrin gene in the C. elegans genome. We show that csq-1 is highly expressed in the body-wall muscles, beginning in mid-embryogenesis and maintained through the adult stage. In body wall muscle cells, CSQ-1 is localized to sarcoplasmic membranes surrounding sarcomeric structures, in the regions where ryanodine receptors (UNC-68) are located. Mutation in UNC-68 affects CSQ-1 localization, suggesting that the two possibly interact in vivo. Genetic analyses of chromosomal deficiency mutants deleting csq-1 show that CSQ-1 is not essential for initiation of embryonic muscle formation and contraction. Furthermore, double-stranded RNA injection resulted in animals completely lacking CSQ-1 in body-wall muscles with no observable defects in locomotion. These findings suggest that although CSQ-1 is one of the major calcium-binding proteins in the body-wall muscles of C. elegans, it is not essential for body-wall muscle formation and contraction. PMID- 11058083 TI - Expression-site-associated gene 8 (ESAG8) of Trypanosoma brucei is apparently essential and accumulates in the nucleolus. AB - Trypanosoma brucei variant surface glycoprotein expression sites are interesting examples of genomic loci under complex epigenetic control. In the infectious bloodstream stage, only one of about 20 expression sites is actively transcribed. In the Tsetse midgut (procyclic) stage, chromatin remodeling silences all expression sites. We have begun to explore the function of one of the expression site-associated genes, ESAG8. Gene knockout experiments implied that ESAG8 is essential. ESAG8 is present at a very low level and apparently accumulates in the nucleolus. A 32-amino-acid domain, which contains a putative bipartite nuclear localization signal (NLS), is both necessary and sufficient to target fusions of ESAG8, with Aequorea victoria green fluorescent protein, to the trypanosome nucleolus. This same sequence functioned only as an NLS in mammalian cells, supporting the idea that nucleolar accumulation requires specific interactions. These results have implications for models of ESAG8 function. PMID- 11058084 TI - Analysis of targeting sequences demonstrates that trafficking to the Toxoplasma gondii plastid branches off the secretory system. AB - Apicomplexan parasites possess a plastid-like organelle called the apicoplast. Most proteins in the Toxoplasma gondii apicoplast are encoded in the nucleus and imported post-translationally. T. gondii apicoplast proteins often have a long N terminal extension that directs the protein to the apicoplast. It can be modeled as a bipartite targeting sequence that contains a signal sequence and a plastid transit peptide. We identified two nuclearly encoded predicted plastid proteins and made fusions with green fluorescent protein to study protein domains required for apicoplast targeting. The N-terminal 42 amino acids of the apicoplast ribosomal protein S9 directs secretion of green fluorescent protein, indicating that targeting to the apicoplast proceeds through the secretory system. Large sections of the S9 predicted transit sequence can be deleted with no apparent impact on the ability to direct green fluorescent protein to the apicoplast. The predicted transit peptide domain of the S9 targeting sequence directs protein to the mitochondrion in vivo. The transit peptide can also direct import of green fluorescent protein into chloroplasts in vitro. These data substantiate the model that protein targeting to the apicoplast involves two distinct mechanisms: the first involving the secretory system and the second sharing features with typical chloroplast protein import. PMID- 11058085 TI - Adhesion-dependent control of matrix metalloproteinase-2 activation in human capillary endothelial cells. AB - The growth and regression of capillary blood vessels during angiogenesis is greatly influenced by changes in the activity of matrix metalloproteinases (MMPs), which selectively degrade extracellular matrix (ECM) and thereby modulate capillary endothelial cell shape, growth and viability. However, changes in cell ECM binding and cell spreading have also been reported to alter MMP secretion and activation. Studies were carried out to determine whether changes in integrin binding or cell shape feed back to alter MMP-2 processing in human capillary endothelial (HCE) cells. Catalytic processing of proMMP-2 to active MMP-2 progressively decreased when HCE cells were cultured on dishes coated with increasing densities of fibronectin (FN), which promote both integrin binding and cell spreading. Conversely, the highest levels of active MMP-2 were detected in round cells cultured on low FN. When measured 24 hours after plating, this increase in active MMP-2 was accompanied by a concomitant rise in mRNA and protein levels for the membrane-type 1 MMP (MT1-MMP), which catalyzes the cleavage of proMMP-2. To determine whether proMMP-2 processing was controlled directly by integrin binding or indirectly by associated changes in cell shape, round cells on low FN were allowed to bind to microbeads (4.5 microm diameter) coated with a synthetic RGD peptide or FN; these induce local integrin receptor clustering without altering cell shape. ProMMP-2 activation was significantly decreased within minutes after bead binding in these round cells, prior to any detectable changes in expression of MT1-MMP, whereas binding of beads coated with control ligands for other transmembrane receptors had no effect. This inhibitory effect was mimicked by microbeads coated with activating antibodies against alphaVbeta3 and beta1 integrins, suggesting a direct role for these cell-surface ECM receptors in modulating proMMP-2 activation. Similar inhibition of proMMP-2 processing by integrin binding, independent of cell spreading, was demonstrated in cells that were cultured on small, microfabricated adhesive islands that prevented cell spreading while presenting a high FN density directly beneath the cell. Interestingly, when spread cells were induced to round up from within by disrupting their actin cytoskeleton using cytochalasin D, proMMP-2 processing did not change at early times; however, increases in MT1-MMP mRNA levels and MMP-2 activation could be detected by 18 hours. Taken together, these results suggest the existence of two phases of MMP-2 regulation in HCE cells when they adhere to ECM: (1) a quick response, in which integrin clustering alone is sufficient to rapidly inhibit processing of proMMP-2 and (2) a slower response, in which subsequent cell spreading and changes in the actin cytoskeleton feed back to decrease expression of MT1-MMP mRNA and, thereby, further suppress cellular proteolytic activity. PMID- 11058086 TI - High dosage expression of a zinc finger protein, Grt1, suppresses a mutant of fission yeast slp1(+), a homolog of CDC20/p55CDC/Fizzy. AB - Selective proteolysis at and after the onset of anaphase is a key cell cycle event required for sister chromatid separation as well as for exit from mitosis. It requires ubiquitination of substrates by Anaphase Promoting Complex(APC)/Cyclosome. Slp1, a WD-repeat protein, is a putative activator for APC in fission yeast. With another WD- repeat protein, Ste9/Srw1, it is thought to promote the proteolysis in a substrate-specific manner. We report here characterization of a temperature-sensitive (ts) slp1 mutant and its high-dosage suppressor, grt1(+). In cells arrested in metaphase, wild-type Slp1 was preferentially found in a complex with hyperphosphorylated Cut9 (subunit of APC), whereas the ts Slp1 protein, lacking the last 113 amino acids, failed to interact with Cut9. The temperature sensitivity was suppressed by high dosage expression of a zinc finger protein, Grt1. The ts slp1 mutant was unable to maintain the normal level of Grt1 protein. The reduction in the Grt1 level may be a primary defect since high dosage expression of grt1(+) rescues the slp1 mutant. The grt1 suppression had an additive effect to ste9 and wee1-50, both of which partially suppress the ts slp1 mutant. Therefore, grt1(+) would define an independent pathway that facilitates the function of Slp1. PMID- 11058087 TI - Differential assembly of alpha- and gamma-filagenins into thick filaments in Caenorhabditis elegans. AB - Muscle thick filaments are highly organized supramolecular assemblies of myosin and associated proteins with lengths, diameters and flexural rigidities characteristic of their source. The cores of body wall muscle thick filaments of the nematode Caenorhabditis elegans are tubular structures of paramyosin sub filaments coupled by filagenins and have been proposed to serve as templates for the assembly of native thick filaments. We have characterized alpha- and gamma filagenins, two novel proteins of the cores with calculated molecular masses of 30,043 and 19,601 and isoelectric points of 10.52 and 11.49, respectively. Western blot and immunoelectron microscopy using affinity-purified antibodies confirmed that the two proteins are core components. Immunoelectron microscopy of the cores revealed that they assemble with different periodicities. Immunofluorescence microscopy showed that alpha-filagenin is localized in the medial regions of the A-bands of body wall muscle cells whereas gamma-filagenin is localized in the flanking regions, and that alpha-filagenin is expressed in 1.5-twofold embryos while gamma-filagenin becomes detectable only in late vermiform embryos. The expression of both proteins continues throughout later stages of development. C. elegans body wall muscle thick filaments of these developmental stages have distinct lengths. Our results suggest that the differential assembly of alpha- and gamma-filagenins into thick filaments of distinct lengths may be developmentally regulated. PMID- 11058088 TI - The Dlx3 protein harbors basic residues required for nuclear localization, transcriptional activity and binding to Msx1. AB - The murine Dlx3 protein is a putative transcriptional activator that has been implicated during development and differentiation of epithelial tissue. Dlx3 contains a homeodomain and mutational analysis has revealed two regions, one N terminal and one C-terminal to the homeodomain, that act as transcriptional activators in a yeast one-hybrid assay. In addition to transactivation, data are presented to demonstrate specific DNA binding and an association between Dlx3 and the Msx1 protein in vitro. Immunohistochemical analysis confirmed coexpression of Dlx3 and Msx1 proteins in the differentiated layers of murine epidermal tissues. Transcription factor function requires nuclear localization. In this study, the intracellular localization of the green fluorescent protein fused to Dlx3 was examined in keratinocytes induced to differentiate by calcium and is shown to localize to the nucleus. A bipartite nuclear localization signal (NLS) was identified by mutational analysis and shown to be sufficient for nuclear localization. This was demonstrated by insertion of the Dlx3 bipartite NLS sequence into a cytoplasmic fusion protein, GFP-keratin 14, which functionally redirected GFP-keratin 14 expression to the nucleus. Further analysis of Dlx3 NLS mutants revealed that the Dlx3 NLS sequences are required for specific DNA binding, transactivation potential and interactions with the Msx1 protein. PMID- 11058089 TI - The breakdown of autophagic vesicles inside the vacuole depends on Aut4p. AB - Autophagy is a degradative transport pathway that delivers cytosolic proteins to the lysosome (vacuole). Cytosolic proteins appear inside the vacuole enclosed in autophagic vesicles. These autophagic vesicles are broken down in the vacuole together with their cytosolic content. The breakdown of vesicular transport intermediates is a unique feature of autophagy. We here identify Aut4p, a component essential for the disintegration of autophagic vesicles, inside the vacuole of S. cerevisiae cells. Aut4p is a putative integral membrane protein with limited homologies to permeases. Chromosomal deletion of AUT4 has no obvious influence on growth, vacuolar acidification and the activities of vacuolar proteinases. Like proteinase B-deficient cells, aut4-deleted cells show a partial reduction in total protein breakdown during nitrogen starvation. A biologically active fusion protein of Aut4p and the green fluorescent protein is visualized at the vacuolar membrane and in punctate structures attached to the vacuole. PMID- 11058090 TI - Processing and trafficking of cysteine proteases in Leishmania mexicana. AB - Removal of the pro-domain of a cysteine protease is essential for activation of the enzyme. We have engineered a cysteine protease (CPB2.8) of the protozoan parasite Leishmania mexicana by site-directed mutagenesis to remove the active site cysteine (to produce CPB(C25G)). When CPB(C25G) was expressed in a L. mexicana mutant lacking all CPB genes, the inactive pro-enzyme was processed to the mature protein and trafficked to the lysosome. These results show that auto activation is not required for correct processing of CPB in vivo. When CPB(C25G) was expressed in a L. mexicana mutant lacking both CPA and CPB genes, the majority of the pro-enzyme remained unprocessed and accumulated in the flagellar pocket. These data reveal that CPA can directly or indirectly process CPB(C25G) and suggest that cysteine proteases are targeted to lysosomes via the flagellar pocket. Moreover, they show that another protease can process CPB in the absence of either CPA or CPB, albeit less efficiently. Abolition of the glycosylation site in the mature domain of CPB did not affect enzyme processing, targeting or in vitro activity towards gelatin. This indicates that glycosylation is not required for trafficking. Together these findings provide evidence that the major route of trafficking of Leishmania cysteine proteases to lysosomes is via the flagellar pocket and therefore differs significantly from cysteine protease trafficking in mammalian cells. PMID- 11058091 TI - Structure and dynamics of hnRNP-labelled nuclear bodies induced by stress treatments. AB - We have previously described HAP, a novel hnRNP protein that is identical both to SAF-B, a component of the nuclear scaffold, and to HET, a transcriptional regulator of the gene for heat shock protein 27. After heat shock, HAP is recruited to a few nuclear bodies. Here we report the characterisation of these bodies, which are distinct from other nuclear components such as coiled bodies and speckles. The formation of HAP bodies is part of a general cell response to stress agents, such as heat shock and cadmium sulfate, which also affect the distribution of hnRNP protein M. Electron microscopy demonstrates that in untreated cells, similar to other hnRNP proteins, HAP is associated to perichromatin fibrils. Instead, in heat shocked cells the protein is preferentially associated to clusters of perichromatin granules, which correspond to the HAP bodies observed in confocal microscopy. Inside such clusters, perichromatin granules eventually merge into a highly packaged 'core'. HAP and hnRNP M mark different districts of these structures. HAP is associated to perichromatin granules surrounding the core, while hnRNP M is mostly detected within the core. BrU incorporation experiments demonstrate that no transcription occurs within the stress-induced clusters of perichromatin granules, which are depots for RNAs synthesised both before and after heat shock. PMID- 11058092 TI - SNAP-24, a Drosophila SNAP-25 homologue on granule membranes, is a putative mediator of secretion and granule-granule fusion in salivary glands. AB - Fusion of vesicles with target membranes is dependent on the interaction of target (t) and vesicle (v) SNARE (soluble NSF (N-ethylmaleimide-sensitive fusion protein) attachment protein receptor) proteins located on opposing membranes. For fusion at the plasma membrane, the t-SNARE SNAP-25 is essential. In Drosophila, the only known SNAP-25 isoform is specific to neuronal axons and synapses and additional t-SNAREs must exist that mediate both non-synaptic fusion in neurons and constitutive and regulated fusion in other cells. Here we report the identification and characterization of SNAP-24, a closely related Drosophila SNAP 25 homologue, that is expressed throughout development. The spatial distribution of SNAP-24 in the nervous system is punctate and, unlike SNAP-25, is not concentrated in synaptic regions. In vitro studies, however, show that SNAP-24 can form core complexes with syntaxin and both synaptic and non-synaptic v SNAREs. High levels of SNAP-24 are found in larval salivary glands, where SNAP-24 localizes mainly to granule membranes rather than the plasma membrane. During glue secretion, the massive exocytotic event of these glands, SNAP-24 containing granules fuse with one another and the apical membrane, suggesting that glue secretion utilizes compound exocytosis and that SNAP-24 mediates secretion. PMID- 11058093 TI - Demonstration of insulin-responsive trafficking of GLUT4 and vpTR in fibroblasts. AB - Insulin-responsive trafficking of the GLUT4 glucose transporter and the insulin regulated aminopeptidase (IRAP) in adipose and muscle cells is well established. Insulin regulation of GLUT4 trafficking in these cells underlies the role that adipose tissue and muscle play in the maintenance of whole body glucose homeostasis. GLUT4 is expressed in a very limited number of tissues, most highly in adipose and muscle, while IRAP is expressed in many tissues. IRAP's physiological role in any of the tissues in which it is expressed, however, is unknown. The fact that IRAP, which traffics by the same insulin-regulated pathway as GLUT4, is expressed in 'non-insulin responsive' tissues raises the question of whether these other cell types also have a specialized insulin-regulated trafficking pathway. The existence of an insulin-responsive pathway in other cell types would allow regulation of IRAP activity at the plasma membrane as a potentially important physiological function of insulin. To address this question we use reporter molecules for both GLUT4 and IRAP trafficking to measure insulin stimulated translocation in undifferentiated cells by quantitative fluorescence microscopy. One reporter (vpTR), a chimera between the intracellular domain of IRAP and the extracellular and transmembrane domains of the transferrin receptor, has been previously characterized. The other is a GLUT4 construct with an exofacial HA epitope and a C-terminal GFP. By comparing these reporters to the transferrin receptor, a marker for general endocytic trafficking, we demonstrate the existence of a specialized, insulin-regulated trafficking pathway in two undifferentiated cell types, neither of which normally express GLUT4. The magnitude of translocation in these undifferentiated cells (approximately threefold) is similar to that reported for the translocation of GLUT4 in muscle cells. Thus, undifferentiated cells have the necessary retention and translocation machinery for an insulin response that is large enough to be physiologically important. PMID- 11058094 TI - Defective organellar membrane protein trafficking in Ap3b1-deficient cells. AB - AP-3 is a heterotetrameric protein complex involved in intracellular vesicle transport. Molecular analyses show that Ap3b1, which encodes the AP-3 (&bgr;)3A subunit, is altered in pearl mice. To provide genetic evidence that mutation of Ap3b1 is responsible for the pearl phenotype and to determine the null phenotype, the Ap3b1 gene was disrupted by homologous recombination. Mice homozygous for the resulting allele, Ap3b1(LN), or compound heterozygotes with pearl, displayed phenotypes similar to those of pearl mice, confirming that Ap3b1 is the causal gene for pearl. Moreover, pearl is likely to be a hypomorph as the Ap3b1(LN) homozygotes had a lighter coat color and accumulated fewer of the micro3 and (&dgr;)3 subunits of AP-3 than did pearl mice. Finally, immunofluorescence analysis of fibroblasts and melanocytes cultured from Ap3b1(LN) homozygotes revealed that the lysosomal membrane proteins Lamp I and Lamp II and the melanosomal membrane protein tyrosinase were mislocalized. In particular, the Lamp proteins were clustered on the cell surface. These findings strengthen the evidence for an alternate pathway via the plasma membrane for cargo normally transported to organelles by AP-3. PMID- 11058095 TI - Involvement of Rb family proteins, focal adhesion proteins and protein synthesis in senescent morphogenesis induced by hydrogen peroxide. AB - Early passage human diploid fibroblasts develop senescent morphology prematurely within a week after a 2-hour pulse treatment with low or mild dose H(2)O(2). We test here the role of cell cycle checkpoints, cytoskeletal proteins and de novo protein synthesis in senescent morphogenesis following H(2)O(2) treatment. H(2)O(2) treatment causes transient elevation of p53 protein and prolonged inhibition of Rb hyperphosphorylation. Expression of human papillomaviral E6 gene prevented elevation of p53 but did not affect senescent morphogenesis. Expression of human papillomaviral E7 gene reduced the level of Rb protein and prevented induction of senescent morphology by H(2)O(2). The mutants of the E7 gene, in which the Rb family protein binding site was destroyed, could not reduce Rb protein or prevent H(2)O(2) from inducing senescent morphology. Senescent-like cells showed enhanced actin stress fibers. In untreated cells, vinculin and paxillin preferentially distributed along the edge of the cells. In contrast, vinculin and paxillin distributed randomly and sporadically throughout senescent like cells. E7 expression prevented enhancement of actin filament formation and redistribution of vinculin or paxillin. Neither wild-type nor E7 cells showed changes in the protein level of actin, vinculin or paxillin measured by western blot after H(2)O(2) treatment. Finally, depletion of methionine in the culture medium after H(2)O(2) treatment prevented senescent morphogenesis without affecting dephosphorylation of Rb protein. Our results suggest that senescent morphology likely develops by a program involving activated Rb family proteins, enhancement of actin stress fibers, redistribution of focal adhesion proteins and de novo protein synthesis. PMID- 11058096 TI - Loss of cation-independent mannose 6-phosphate receptor expression promotes the accumulation of lysobisphosphatidic acid in multilamellar bodies. AB - A number of recent studies have highlighted the importance of lipid domains within endocytic organelles in the sorting and movement of integral membrane proteins. In particular, considerable attention has become focussed upon the role of the unusual phospholipid lysobisphosphatidic acid (LBPA). This lipid appears to be directly involved in the trafficking of cholesterol and glycosphingolipids, and accumulates in a number of lysosomal storage disorders. Antibody-mediated disruption of LBPA function also leads to mis-sorting of cation-independent mannose 6-phosphate receptors. We now report that the converse is also true, and that spontaneous loss of cation-independent mannose 6-phosphate receptors from a rat fibroblast cell line led to the formation of aberrant late endocytic structures enriched in LBPA. Accumulation of LBPA was directly dependent upon the loss of the receptors, and could be reversed by expression of bovine cation independent mannose 6-phosphate receptors in the mutant cell line. Ultrastructural analysis indicated that the abnormal organelles were electron dense, had a multi-lamellar structure, accumulated endocytosed probes, and were distinct from dense-core lysosomes present within the same cells. The late endocytic structures present at steady state within any particular cell likely reflect the balance of membrane traffic through the endocytic pathway of that cell, and the rate of maturation of individual endocytic organelles. Moreover, there is considerable evidence which suggests that cargo receptors also play a direct mechanistic role in membrane trafficking events. Therefore, loss of such a protein may disturb the overall equilibrium of the pathway, and hence cause the accumulation of aberrant organelles. We propose that this mechanism underlies the phenotype of the mutant cell line, and that the formation of inclusion bodies in many lysosomal storage diseases is also due to an imbalance in membrane trafficking within the endocytic pathway. PMID- 11058097 TI - Connexin-specific distribution within gap junctions revealed in living cells. AB - To study the organization of gap junctions in living cells, the connexin isotypes alpha(1)(Cx43), beta(1)(Cx32) and beta(2)(Cx26) were tagged with the autofluorescent tracer green fluorescent protein (GFP) and its cyan (CFP) and yellow (YFP) color variants. The cellular fate of the tagged connexins was followed by high-resolution fluorescence deconvolution microscopy and time-lapse imaging. Comprehensive analyses demonstrated that the tagged channels were functional as monitored by dye transfer, even under conditions where the channels were assembled solely from tagged connexins. High-resolution images revealed a detailed structural organization, and volume reconstructions provided a three dimensional view of entire gap junction plaques. Specifically, deconvolved dual color images of gap junction plaques assembled from CFP- and YFP-tagged connexins revealed that different connexin isotypes gathered within the same plaques. Connexins either codistributed homogeneously throughout the plaque, or each connexin isotype segregated into well-separated domains. The studies demonstrate that the mode of channel distribution strictly depends on the connexin isotypes. Based on previous studies on the synthesis and assembly of connexins I suggest that channel distribution is regulated by intrinsic connexin isotype specific signals. PMID- 11058098 TI - The armadillo repeat region targets ARVCF to cadherin-based cellular junctions. AB - The cytoplasmic domain of the transmembrane protein M-cadherin is involved in anchoring cytoskeletal elements to the plasma membrane at cell-cell contact sites. Several members of the armadillo repeat protein family mediate this linkage. We show here that ARVCF, a member of the p120 (ctn) subfamily, is a ligand for the cytoplasmic domain of M-cadherin, and characterize the regions involved in this interaction in detail. Complex formation in an in vivo environment was demonstrated in (1) yeast two-hybrid screens, using a cDNA library from differentiating skeletal muscle and part of the cytoplasmic M cadherin tail as a bait, and (2) mammalian cells, using a novel experimental system, the MOM recruitment assay. Immunoprecipitation and in vitro binding assays confirmed this interaction. Ectopically expressed EGFP-ARVCF-C11, an N terminal truncated fragment, targets to junctional structures in epithelial MCF7 cells and cardiomyocytes, where it colocalizes with the respective cadherins, beta-catenin and p120 (ctn). Hence, the N terminus of ARVCF is not required for junctional localization. In contrast, deletion of the four N-terminal armadillo repeats abolishes this ability in cardiomyocytes. Detailed mutational analysis revealed the armadillo repeat region of ARVCF as sufficient and necessary for interaction with the 55 membrane-proximal amino acids of the M-cadherin tail. PMID- 11058099 TI - Structural and mechanistic conservation in DNA ligases. AB - DNA ligases are enzymes required for the repair, replication and recombination of DNA. DNA ligases catalyse the formation of phosphodiester bonds at single-strand breaks in double-stranded DNA. Despite their occurrence in all organisms, DNA ligases show a wide diversity of amino acid sequences, molecular sizes and properties. The enzymes fall into two groups based on their cofactor specificity, those requiring NAD(+) for activity and those requiring ATP. The eukaryotic, viral and archael bacteria encoded enzymes all require ATP. NAD(+)-requiring DNA ligases have only been found in prokaryotic organisms. Recently, the crystal structures of a number of DNA ligases have been reported. It is the purpose of this review to summarise the current knowledge of the structure and catalytic mechanism of DNA ligases. PMID- 11058100 TI - HIV-1 LTR as a target for synthetic ribozyme-mediated inhibition of gene expression: site selection and inhibition in cell culture. AB - A library of three synthetic ribozymes with randomized arms, targeting NUX, GUX and NXG triplets, respectively, were used to identify ribozyme-accessible sites on the HIV-1 LTR transcript comprising positions -533 to 386. Three cleavable sites were identified at positions 109, 115 and 161. Ribozymes were designed against these sites, either unmodified or with 2'-modifications and phosphorothioate groups, and their cleavage activities of the transcript were determined. Their biological activities were assessed in cell culture, using a HIV-1 model assay system where the LTR is a promoter for the expression of the reporter gene luciferase in a transient expression system. Intracellular efficiency of the ribozymes were determined by cotransfection of ribozyme and plasmid DNA, expressing the target RNA. Modified ribozymes, directed against positions 115 and 161, lowered the level of LTR mRNA in the cell resulting in inhibition of expression of the LTR-driven reporter gene luciferase of 87 and 61%, respectively. In the presence of Tat the inhibitions were 43 and 25%. The inactive variants of these ribozymes exhibited a similar inhibitory effect. RNase protection revealed a reduction of RNA which was somewhat stronger for the active than the inactive ribozymes, particularly for ribozyme 115. Unmodified ribozymes showed no inhibition in the cell. The third ribozyme, targeting a GUG-triplet at position 109, possessed only low cleavage activity in vitro and no inhibitory effect in cell culture. PMID- 11058101 TI - Biochemical characterisation of cap-poly(A) synergy in rabbit reticulocyte lysates: the eIF4G-PABP interaction increases the functional affinity of eIF4E for the capped mRNA 5'-end. AB - The 5' cap and 3' poly(A) tail of eukaryotic mRNAs cooperate to synergistically stimulate translation initiation in vivo. We recently described mammalian cytoplasmic extracts which, following ultracentrifugation to partially deplete them of ribosomes and associated initiation factors, reproduce cap-poly(A) synergy in vitro. Using these systems, we demonstrate that synergy requires interaction between the poly(A)-binding protein (PABP) and the eukaryotic initiation factor (eIF) 4F holoenzyme complex, which recognises the 5' cap. Here we further characterise the requirements and constraints of cap-poly(A) synergy in reticulocyte lysates by evaluating the effects of different parameters on synergy. The extent of extract depletion and the amounts of different initiation factors in depleted extracts were examined, as well as the effects of varying the concentrations of KCl, MgCl(2) and programming mRNA and of adding a cap analogue. The results presented demonstrate that maximal cap-poly(A) synergy requires: (i) limiting concentrations of ribosome-associated initiation factors; (ii) precise ratios of mRNA to translation machinery (low concentrations of ribosome associated initiation factors and low, non-saturating mRNA concentrations); (iii) physiological concentrations of added KCl and MgCl(2). Additionally, we show that the eIF4G-PABP interaction on mRNAs which are capped and polyadenylated significantly increases the affinity of eIF4E for the 5' cap. PMID- 11058102 TI - DNA binding properties of the Arabidopsis floral development protein AINTEGUMENTA. AB - The Arabidopsis protein AINTEGUMENTA (ANT) is a member of a plant-specific family of transcription factors (AP2/EREBP) that share either one or two copies of an approximately 70 amino acid region called the AP2 repeat. DNA binding activity has been demonstrated previously for members of this family containing a single AP2 repeat. Using an in vitro selection procedure, the DNA binding specificity of the two AP2 repeat containing protein ANT was found to be 5' gCAC(A/G)N(A/T)TcCC(a/g)ANG(c/t)-3'. This consensus site is much longer than sites recognized by proteins containing a single AP2 repeat and neither AP2 repeat of ANT was alone capable of binding to the selected sequences, suggesting that both AP2 repeats make DNA contacts. ANT binds to these DNA sequences as a monomer but a higher order complex is also observed at high protein concentrations. The ANT consensus site shows some similarity to the C-repeat/DRE elements bound by proteins that contain a single AP2 repeat, and we find that ANT binds weakly to such sites. We propose a model in which each AP2 repeat of ANT contacts adjacent sites within the consensus sequence. Our results suggest that the AP2 repeat can be utilized in different ways for DNA binding. PMID- 11058103 TI - Poly(dA.dT) sequences exist as rigid DNA structures in nucleosome-free yeast promoters in vivo. AB - Poly(dA.dT) sequences (T-tracts) are abundant genomic DNA elements with unusual properties in vitro and an established role in transcriptional regulation of yeast genes. In vitro T-tracts are rigid, contribute to DNA bending, affect assembly in nucleosomes and generate a characteristic pattern of CPDs (cyclobutane pyrimidine dimers) upon irradiation with UV light (UV photofootprint). In eukaryotic cells, where DNA is packaged in chromatin, the DNA structure of T-tracts is unknown. Here we have used in vivo UV photofootprinting and DNA repair by photolyase to investigate the structure and accessibility of T tracts in yeast promoters (HIS3, URA3 and ILV1). The same characteristic photofootprints were obtained in yeast and in naked DNA, demonstrating that the unusual T-tract structure exists in living cells. Rapid repair of CPDs in the T tracts demonstrates that these T-tracts were not folded in nucleosomes. Moreover, neither datin, a T-tract binding protein, nor Gcn5p, a histone acetyltransferase involved in nucleosome remodelling, showed an influence on the structure and accessibility of T-tracts. The data support a contribution of this unusual DNA structure to transcriptional regulation. PMID- 11058104 TI - The yeast CDP1 gene encodes a triple-helical DNA-binding protein. AB - The formation of triple-helical DNA has been implicated in several cellular processes, including transcription, replication and recombination. While there is no direct evidence for triplexes in vivo, cellular proteins that specifically recognize triplex DNA have been described. Using a purine-motif triplex probe and southwestern library screening, we isolated five independent clones expressing the same C-terminal 210 amino acids of the Saccharomyces cerevisiae protein Cdp1p fused with beta-galactosidase. In electrophoretic mobility shift assays, recombinant Cdp1pDelta1-867 bound Pu-motif triplex DNAs with high affinity (K:(d) approximately 5 nM) and bound Py-motif triplex, duplex and single-stranded DNAs with far lower affinity (0.5-5.0 microM). Genetic analyses revealed that the CDP1 gene product was required for proper chromosome segregation. The possible involvement of triplex DNA in this process is discussed. PMID- 11058105 TI - Purification and cloning of cytotoxic ribonucleases from Rana catesbeiana (bullfrog). AB - Ribonucleases with antitumor activity are mainly found in the oocytes and embryos of frogs, but the role of these ribonucleases in frog development is not clear. Moreover, most frog ribonuclease genes have not been cloned and characterized. In the present study, a group of ribonucleases were isolated from Rana catesbeiana (bullfrog). These ribonucleases in mature oocytes, namely RC-RNase, RC-RNase 2, RC-RNase 3, RC-RNase 4, RC-RNase 5 and RC-RNase 6, as well as liver-specific ribonuclease RC-RNase L1, were purified by column chromatographs and detected by zymogram assay and western blotting. Characterization of these purified ribonucleases revealed that they were highly conserved in amino acid sequence and had a pyroglutamate residue at their N-termini, but possessed different specific activities, base specificities and optimal pH values for their activities. These ribonucleases were cytotoxic to cervical carcinoma HeLa cells, but their cytotoxicities were not closely correlated to their enzymatic specific activities. Some other amino acid residues in addition to their catalytic residues were implicated to be involved in the cytotoxicity of the frog ribonucleases to tumor cells. Because the coding regions lack introns, the ribonuclease genes were cloned by PCR using genomic DNA as template. Their DNA sequences and amino acid sequences are homologous to those of mammalian ribonuclease superfamily, approximately 50 and approximately 25%, respectively. PMID- 11058106 TI - A family of developmentally excised DNA elements in Tetrahymena is under selective pressure to maintain an open reading frame encoding an integrase-like protein. AB - Tlr1 is a member of a family of approximately 20-30 DNA elements that undergo developmentally regulated excision during formation of the macronucleus in the ciliated protozoan TETRAHYMENA: Analysis of sequence internal to the right boundary of Tlr1 revealed the presence of a 2 kb open reading frame (ORF) encoding a deduced protein with similarity to retrotransposon integrases. The ORFs of five unique clones were sequenced. The ORFs have 98% sequence conservation and align without frameshifts, although one has an additional trinucleotide at codon 561. Nucleotide changes among the five clones are highly non-random with respect to the position in the codon and 93% of the nucleotide changes among the five clones encode identical or similar amino acids, suggesting that the ORF has evolved under selective pressure to preserve a functional protein. Nineteen T/C transitions in T/CAA and T/CAG codons suggest selection has occurred in the context of the TETRAHYMENA: genome, where TAA and TAG encode Gln. Similarities between the ORF and those encoding retrotransposon integrases suggest that the Tlr family of elements may encode a polynucleotide transferase. Possible roles for the protein in transposition of the elements within the micronuclear genome and/or their developmentally regulated excision from the macronucleus are discussed. PMID- 11058107 TI - Secondary structure prediction and in vitro accessibility of mRNA as tools in the selection of target sites for ribozymes. AB - We have investigated the relative merits of two commonly used methods for target site selection for ribozymes: secondary structure prediction (MFold program) and in vitro accessibility assays. A total of eight methylated ribozymes with DNA arms were synthesized and analyzed in a transient co-transfection assay in HeLa cells. Residual expression levels ranging from 23 to 72% were obtained with anti PSKH1 ribozymes compared to cells transfected with an irrelevant control ribozyme. Ribozyme efficacy depended on both ribozyme concentration and the steady state expression levels of the target mRNA. Allylated ribozymes against a subset of the target sites generally displayed poorer efficacy than their methylated counterparts. This effect appeared to be influenced by in vivo accessibility of the target site. Ribozymes designed on the basis of either selection method displayed a wide range of efficacies with no significant differences in the average activities of the two groups of ribozymes. While in vitro accessibility assays had limited predictive power, there was a significant correlation between certain features of the predicted secondary structure of the target sequence and the efficacy of the corresponding ribozyme. Specifically, ribozyme efficacy appeared to be positively correlated with the presence of short stem regions and helices of low stability within their target sequences. There were no correlations with predicted free energy or loop length. PMID- 11058108 TI - Conformation of oligodeoxynucleotides associated with anionic liposomes. AB - There has been significant progress in the development of antisense therapeutics for a wide range of medicinal applications. Further improvement will require better understanding of cellular internalization, intracellular distribution mechanisms and interactions of oligodeoxynucleotides with cellular organelles. In many of these processes interactions of oligodeoxynucleotides with lipid assemblies may have a significant influence on their function. Divalent cations have been shown to assist cellular internalization of certain oligodeoxynucleotides and to affect their conformation. In this work we have investigated conformational changes of phosphorothioate oligodeoxynucleotides upon divalent cation-mediated interaction with 1,2-dipalmitoyl-sn-glycero-3 phosphatidylglycerol (DPPG) liposomes. For the sequences investigated here the native conformation underwent significant change in the presence of anionic DPPG liposomes only when divalent cations were present. This change is sequence specific, ion-selective and distinct from previously reported changes in oligodeoxynucleotide structure due to divalent cations alone. The conformation of one oligodeoxynucleotide in the presence of calcium and DPPG yields circular dichroism spectra which suggest C-DNA but which also have characteristics unlike any previously reported spectra of liposome-associated DNA structure. The data suggest the possibility of a unique conformation of liposome-associated ODNs and reflect a surprisingly strong tendency of single-stranded DNA to retain a characteristic conformation even when adsorbed to a surface. This conformation may be related to cellular uptake, transport of oligodeoxynucleotides in cells and/or function. PMID- 11058109 TI - Stabilization of the U5-leader stem in the HIV-1 RNA genome affects initiation and elongation of reverse transcription. AB - Reverse transcription of the Human Immunodeficiency Virus type I (HIV-1) RNA genome is primed by a cellular tRNA-lys3 molecule that binds to the primer binding site (PBS). The PBS is predicted to be part of an extended RNA structure, consisting of a small U5-PBS hairpin and a large U5-leader stem. In this study we stabilized the U5-leader stem of HIV-1 to study its role in reverse transcription. We tested in vitro synthesized wild-type and mutant templates in primer annealing, initiation and elongation assays. Stabilization of the stem inhibits the initiation of reverse transcription, but not the annealing of the tRNA primer onto the PBS. These results suggest that stabilization of the stem results in occlusion of a sequence motif that is involved in an additional interaction with the tRNA-lys3 primer and that is needed to trigger the initiation of reverse transcription. The stable structure was also found to affect the elongation of reverse transcription, causing the RT enzyme to pause upon copying 7-8 bases into the extended base paired stem. The stabilizing mutations were also introduced into proviral constructs for replication studies, demonstrating that the mutant viruses have a reduced replication capacity. Analysis of a revertant virus demonstrated that opening of the stabilized U5 leader stem can restore both virus replication and reverse transcription. PMID- 11058110 TI - Error-free and error-prone lesion bypass by human DNA polymerase kappa in vitro. AB - Error-free lesion bypass and error-prone lesion bypass are important cellular responses to DNA damage during replication, both of which require a DNA polymerase (Pol). To identify lesion bypass DNA polymerases, we have purified human Polkappa encoded by the DINB1 gene and examined its response to damaged DNA templates. Here, we show that human Polkappa is a novel lesion bypass polymerase in vitro. Purified human Polkappa efficiently bypassed a template 8-oxoguanine, incorporating mainly A and less frequently C opposite the lesion. Human Polkappa most frequently incorporated A opposite a template abasic site. Efficient further extension required T as the next template base, and was mediated mainly by a one nucleotide deletion mechanism. Human Polkappa was able to bypass an acetylaminofluorene-modified G in DNA, incorporating either C or T, and less efficiently A opposite the lesion. Furthermore, human Polkappa effectively bypassed a template (-)-trans-anti-benzo[a]pyrene-N:(2)-dG lesion in an error free manner by incorporating a C opposite the bulky adduct. In contrast, human Polkappa was unable to bypass a template TT dimer or a TT (6-4) photoproduct, two of the major UV lesions. These results suggest that Polkappa plays an important role in both error-free and error-prone lesion bypass in humans. PMID- 11058111 TI - Human DNA polymerase kappa synthesizes DNA with extraordinarily low fidelity. AB - Escherichia coli DNA polymerase IV encoded by the dinB gene is involved in untargeted mutagenesis. Its human homologue is DNA polymerase kappa (Polkappa) encoded by the DINB1 gene. Our recent studies have indicated that human Polkappa is capable of both error-free and error-prone translesion DNA synthesis in vitro. However, it is not known whether human Polkappa also plays a role in untargeted mutagenesis. To examine this possibility, we have measured the fidelity of human Polkappa during DNA synthesis from undamaged templates. Using kinetic measurements of nucleotide incorporations and a fidelity assay with gapped M13mp2 DNA, we show that human Polkappa synthesizes DNA with extraordinarily low fidelity. At the lacZalpha target gene, human Polkappa made on average one error for every 200 nucleotides synthesized, with a predominant T-->G transversion mutation at a rate of 1/147. The overall error rate of human Polkappa is 1.7-fold lower than human Poleta, but 33-fold higher than human Polbeta, a DNA polymerase with very low fidelity. Thus, human Polkappa is one of the most inaccurate DNA polymerases known. These results support a role for human Polkappa in untargeted mutagenesis surrounding a DNA lesion and in DNA regions without damage. PMID- 11058112 TI - 5-Methylcytosine DNA glycosylase activity is also present in the human MBD4 (G/T mismatch glycosylase) and in a related avian sequence. AB - A 1468 bp cDNA coding for the chicken homolog of the human MBD4 G/T mismatch DNA glycosylase was isolated and sequenced. The derived amino acid sequence (416 amino acids) shows 46% identity with the human MBD4 and the conserved catalytic region at the C-terminal end (170 amino acids) has 90% identity. The non conserved region of the avian protein has no consensus sequence for the methylated DNA binding domain. The recombinant proteins from human and chicken have G/T mismatch as well as 5-methylcytosine (5-MeC) DNA glycosylase activities. When tested by gel shift assays, human recombinant protein with or without the methylated DNA binding domain binds equally well to symmetrically, hemimethylated DNA and non-methylated DNA. However, the enzyme has only 5-MeC DNA glycosylase activity with the hemimethylated DNA. Footprinting of human MBD4 and of an N terminal deletion mutant with partially depurinated and depyrimidinated substrate reveal a selective binding of the proteins to the modified substrate around the CpG. As for 5-MeC DNA glycosylase purified from chicken embryos, MBD4 does not use oligonucleotides containing mCpA, mCpT or mCpC as substrates. An mCpG within an A+T-rich oligonucleotide is a much better substrate than an A+T-poor sequence. The K:(m) of human MBD4 for hemimethylated DNA is approximately 10(-7) M with a V:(max) of approximately 10(-11) mol/h/microgram protein. Deletion mutations show that G/T mismatch and 5-MeC DNA glycosylase are located in the C-terminal conserved region. In sharp contrast to the 5-MeC DNA glycosylase isolated from the chicken embryo DNA demethylation complex, the two enzymatic activities of MBD4 are strongly inhibited by RNA. In situ hybridization with antisense RNA indicate that MBD4 is only located in dividing cells of differentiating embryonic tissues. PMID- 11058113 TI - An imperfect inverted repeat is critical for DNA binding of the response regulator RegR of Bradyrhizobium japonicum. AB - RegR is the response regulator of the RegSR two-component regulatory system in Bradyrhizobium japonicum. The only target known so far is the fixR-nifA operon, encoding the redox-responsive transcription factor NifA, which activates many genes required for symbiotic nitrogen fixation in soybean nodules. In previous in vivo studies, we identified a 32 bp upstream activating sequence located around position -68, which is essential for RegR-dependent expression of the fixR-nifA operon. Here, we used an in vitro binding-site selection assay (SELEX) to more precisely define the DNA-binding specificity of RegR. The selected sequences comprised an imperfect inverted repeat (GCGGC-N(5)-GTCGC) which is highly similar to an imperfect inverted repeat in the fixR UAS (GCGAC-N(5)-GACGC). In a parallel approach, band-shift experiments were performed with oligonucleotides comprising defined point or deletion mutations in the fixR UAS. This led to the identification of 11 critical nucleotides within a 17 bp minimal RegR binding site centered at position -64 upstream of the fixR-nifA transcription start site. Notably, all 11 critical nucleotides were located either within the half sites of the inverted repeat (four nucleotides in each half site) or in the 5 bp spacer that separates the half sites (three nucleotides). Based on these results, we defined a DNA motif comprising those nucleotides that are critical for RegR binding (RegR box; 5'-GNG(A)(G)C(A)(G)TTNNGNCGC-3'). A comparison of the RegR box with functional binding sites of the RegR-like regulator RegA of Rhodobacter capsulatus revealed considerable similarities. Thus, the RegR box may assist in the identification of new RegR target genes not only in B.japonicum but also in other alpha-proteobacteria possessing RegR-like response regulators. PMID- 11058114 TI - Functional consequences of Rett syndrome mutations on human MeCP2. AB - The neurodevelopmental disorder known as Rett syndrome has recently been linked to the methyl-CpG-binding transcriptional repressor, MeCP2. In this report we examine the consequences of these mutations on the function of MeCP2. The ability to bind specifically to methylated DNA and the transcription repression capabilities are tested, as well as the stability of proteins in vivo. We find that all missense mutations (R106W, R133C, F155S, T158M) within the methyl binding domain impair selectivity for methylated DNA, and that all nonsense mutations (L138X, R168X, E235X, R255X, R270X, V288X, R294X) that truncate all or some of the transcriptional repression domain (TRD) affect the ability to repress transcription and have decreased levels of stability in vivo. Two missense mutations, one in the TRD (R306C) and one in the C-terminus (E397K), had no noticeable effects on MeCP2 function. Together, these results provide evidence of how Rett syndrome mutations can affect distinct functions of MeCP2 and give insight into these mutations that may contribute to the disease. PMID- 11058115 TI - Proteolysis of the human DNA polymerase epsilon catalytic subunit by caspase-3 and calpain specifically during apoptosis. AB - Human DNA polymerase epsilon (pol epsilon) normally contains a 261-kDa catalytic subunit (p261), but from some sources it is isolated as a 140-kDa catalytic core of p261. This shortened form possesses normal or somewhat enhanced polymerase activity and its significance is unknown. We report here that caspase-3 and calpain can form p140 from p261 in vitro and in vivo and that during early stages of apoptosis induced in Jurkat cells by staurosporine or anti-Fas-activating antibody, p261 is cleaved into p140 by caspase-3. At later stages, activated calpain might also contribute to this conversion. The sites of cleavage by caspase-3 have been identified, and mutations at these 'DEAD boxes' resulted in cleavage-resistant enzyme. Cleavage at these sites separates the 'N-terminal catalytic core' from the 'C-terminal' regions described for p261. Cleavage does not occur during necrosis or following exposure to H(2)O(2) or methanesulfonic acid methyl ester. p140 is unlikely to be able to functionally replace p261 in vivo, since it does not bind to PCNA or the other pol epsilon subunits. PMID- 11058116 TI - Analysis of Groucho-histone interactions suggests mechanistic similarities between Groucho- and Tup1-mediated repression. AB - The Drosophila Groucho (Gro) protein is the defining member of a family of metazoan corepressors that have roles in many aspects of development, including segmentation, dorsal/ventral pattern formation, Notch signaling, and Wnt/Wg signaling. Previous speculation has suggested that Gro may be orthologous to the yeast corepressor Tup1. In support of this idea, a detailed alignment between the C-terminal WD-repeat domains of these two proteins shows that each Gro WD repeat is most similar to the Tup1 WD repeat occupying the corresponding position in that protein. Our analysis of Gro-histone interactions provides further support for a close evolutionary relationship between Gro and Tup1. In particular, we show that, as with the N-terminal region of Tup1, the N-terminal region of Gro is necessary and sufficient for direct binding to histones. The highest affinity interaction is with histone H3 and binding is primarily observed with hypoacetylated histones. Using transient transfection assays, we show that a Gal4 Gro fusion protein containing the histone-binding domain is able to repress transcription. Deletions that weaken histone binding also weaken repression. These findings, along with our recent report that Gro interacts with the histone deacetylase Rpd3, suggest a mechanism for Gro-mediated repression. PMID- 11058117 TI - Quantitative studies of Mn(2+)-promoted specific and non-specific cleavages of a large RNA: Mn(2+)-GAAA ribozymes and the evolution of small ribozymes. AB - Manganese (Mn(2+)) promotes specific cleavage at two major (I and III) and four minor (II, IV, V and VI) sites, in addition to slow non-specific cleavage, in a 659-nucleotide RNA containing the Cr.LSU group I intron. The specific cleavages occurred between G and AAA sequences and thus can be considered Mn(2+)-GAAA ribozymes. We have estimated rates of specific and non-specific cleavages under different conditions. Comparisons of the rates of major-specific and background cleavages gave a maximal specificity of approximately 900 for GAAA cleavage. Both specific and non-specific cleavages showed hyperbolic kinetics and there was no evidence of cooperativity with Mn(2+) concentration. Interestingly, at site III, Mg(2+) alone promoted weak, but the same specific cleavage as Mn(2+). When added with Mn(2+), Mg(2+) had a synergistic effect on cleavage at site III, but inhibited cleavage at the other sites. Mn(2+) cleavage at site III also exhibited lower values of K (Mn(2+) requirement), pH-dependency and activation energy than did cleavage at the other sites. In contrast, the pH-dependency and activation energy for cleavage at site I was similar to non-specific cleavage. These results increase our understanding of the Mn(2+)-GAAA ribozyme. The implications for evolution of small ribozymes are also discussed. PMID- 11058118 TI - Pre-steady state kinetics of bacteriophage T4 dam DNA-[N(6)-adenine] methyltransferase: interaction with native (GATC) or modified sites. AB - The DNA methyltransferase of bacteriophage T4 (T4 Dam MTase) recognizes the palindromic sequence GATC, and catalyzes transfer of the methyl group from S: adenosyl-L-methionine (AdoMet) to the N(6)-position of adenine [generating N(6) methyladenine and S:-adenosyl-L-homocysteine (AdoHcy)]. Pre-steady state kinetic analysis revealed that the methylation rate constant k(meth) for unmethylated and hemimethylated substrates (0.56 and 0.47 s(-1), respectively) was at least 20 fold larger than the overall reaction rate constant k(cat) (0.023 s(-1)). This indicates that the release of products is the rate-limiting step in the reaction. Destabilization of the target-base pair did not alter the methylation rate, indicating that the rate of target nucleoside flipping does not limit k(meth). Preformed T4 Dam MTase-DNA complexes are less efficient than preformed T4 Dam MTase-AdoMet complexes in the first round of catalysis. Thus, this data is consistent with a preferred route of reaction for T4 Dam MTase in which AdoMet is bound first; this preferred reaction route is not observed with the DNA-[C5 cytosine]-MTases. PMID- 11058119 TI - A novel cytoplasmic GTPase XAB1 interacts with DNA repair protein XPA. AB - The xeroderma pigmentosum group A protein (XPA) plays a central role in nucleotide excision repair (NER). To identify proteins that bind to XPA, we screened a HeLa cDNA library using the yeast two-hybrid system. Here we report a novel cytoplasmic GTP-binding protein, designated XPA binding protein 1 (XAB1). The deduced amino acid sequence of XAB1 consisted of 374 residues with a molecular weight of 41 kDa and an isoelectric point of 4.65. Sequence analysis revealed that XAB1 has four sequence motifs G1-G4 of the GTP-binding protein family in the N-terminal half. XAB1 also contains an acidic region in the C terminal portion. Northern blot analysis showed that XAB1 mRNA is expressed ubiquitously, and immunofluorescence analysis revealed that XAB1 is localized mainly in the cytoplasm. Consistent with the GTP-binding motif, purified recombinant XAB1 protein has intrinsic GTPase activity. Using the yeast two hybrid system, we elucidated that XAB1 binds to the N-terminal region of XPA. The deletion of five amino acids, residues 30-34 of XPA, required for nuclear localization of XPA abolished the interaction with XAB1. These results suggest that XAB1 is a novel cytoplasmic GTPase involved in nuclear localization of XPA. PMID- 11058120 TI - Cloning of an interferon regulatory factor 2 isoform with different regulatory ability. AB - Interferons (IFNs) are a family of multifunctional proteins involved in immune activation, regulation of cell growth and antiviral response. They exert their functions by induction of several IFN-stimulated genes, including IFN regulatory factors (IRFs), a family of transcriptional regulators. One of these factors, IRF 2, was initially cloned as an antagonistic counterpart to IRF-1 with oncogenic potential. Here we describe a second isoform of IRF-2, termed IRF-2s, cloned from human and murine cells. This isoform lacks two amino acids located C-terminal of the DNA-binding domain, which is conserved in all IRF family members, leading to a change in the predicted secondary structure. Both isoforms have similar binding affinities to known target sequences in electrophoretic mobility shift assays. Using reporter gene constructs with the type IV promoter region of the MHC class II transactivator (CIITA), which is the essential factor for IFN-gamma-induced MHC class II expression, we show that the short isoform IRF-2s exhibits a weaker activation ability compared to IRF-2. Thus, our data present the first evidence of two IRF-2 isoforms with different regulatory ability. PMID- 11058121 TI - Enhanced delivery of antisense oligonucleotides with fluorophore-conjugated PAMAM dendrimers. AB - PAMAM dendrimers are cationic polymers that have been used for the delivery of genes and oligonucleotides to cells. However, little is known about the behavior of dendrimer-nucleic acid complexes once they reach the cell interior. To pursue this issue, we prepared dendrimers conjugated with the fluorescent dye Oregon green 488. These were used in conjunction with oligonucleotides labeled with a red (TAMRA) fluorophore in order to visualize the sub-cellular distribution of the dendrimer-oligonucleotide complex and of its components by two-color digital fluorescence microscopy. The 2'-O:-methyl antisense oligonucleotide sequence used in these studies was designed to correct splicing at an aberrant intron inserted into a luciferase reporter gene; thus effective delivery of the antisense agent results in the expression of the reporter gene product. The dendrimer oligonucleotide complex remained associated during the process of uptake into vesicular compartments and eventual entry into the nucleus. Since the pharmacological activity of the antisense compound was manifest under these conditions, it suggests that the dendrimer-oligonucleotide complex is functionally active. A surprising result of these studies was that the Oregon green 488-conjugated dendrimer was a much better delivery agent for antisense compounds than unmodified dendrimer. This suggests that coupling of relatively hydrophobic small molecules to PAMAM dendrimers may provide a useful means of enhancing their capabilities as delivery agents for nucleic acids. PMID- 11058122 TI - Hyper-resistance of meiotic cells to radiation due to a strong expression of a single recA-like gene in Caenorhabditis elegans. AB - Sensitivity of meiotic cells to DNA damaging agents is little understood. We have demonstrated that the meiotic pachytene nuclei in the Caenorhabditis elegans gonad are hyper-resistant to X-ray irradiation, but not to UV irradiation, whereas the early embryonic cells after fertilization and the full grown oocytes are not. The Ce-rdh-1 gene [RAD51, DMC1 (LIM15), homolog 1 or Ce-rad-51], which is essential for the meiotic recombination, is the only bacterial recA-like gene in the nematode genome, and is strongly expressed in the meiotic cells. Following silencing of the Ce-rdh-1 gene by RNA interference, the meiotic cells become more sensitive to X-ray irradiation than the early embryonic cells. This is the first report that meiotic cells are hyper-resistant to DNA strand breaks due to the high level of expression of the enzyme(s) involved in meiotic homologous recombination. PMID- 11058123 TI - Formation of cis-diamminedichloroplatinum(II) 1,2-intrastrand cross-links on DNA is flanking-sequence independent. AB - Mapping of cis-diamminedichloroplatinum(II) (cis-DDP, cisplatin) DNA adducts over >3000 nucleotides was carried out using a replication blockage assay. The sites of inhibition of modified T4 DNA polymerase, also referred to as stop sites, were analyzed to determine the effects of local sequence context on the distribution of intrastrand cisplatin cross-links. In a 3120 base fragment from replicative form M13mp18 DNA containing 24.6% guanine, 25.5% thymine, 26.9% adenine and 23.0% cytosine, 166 individual stop sites were observed at a bound platinum/nucleotide ratio of 1-2 per thousand. The majority of stop sites (90%) occurred at G(n>2) sequences and the remainder were located at sites containing an AG dinucleotide. For all of the GG sites present in the mapped sequences, including those with Gn(>)2, 89% blocked replication, whereas for the AG sites only 17% blocked replication. These blockage sites were independent of flanking nucleotides in a sequence of N(1)G*G*N(2) where N(1), N(2) = A, C, G, T and G*G* indicates a 1,2 intrastrand platinum cross-link. The absence of long-range sequence dependence was confirmed by monitoring the reaction of cisplatin with a plasmid containing an 800 bp insert of the human telomere repeat sequence (TTAGGG)(n). Platination reactions monitored at several formal platinum/nucleotide ratios or as a function of time reveal that the telomere insert was not preferentially damaged by cisplatin. Both replication blockage and telomere-insert plasmid platination experiments indicate that cisplatin 1,2-intrastrand adducts do not form preferentially at G-rich sequences in vitro. PMID- 11058124 TI - A novel form of intercalation involving four DNA duplexes in an acridine-4 carboxamide complex of d(CGTACG)(2). AB - The structures of the complexes formed between 9-amino-[N:-(2-dimethyl amino)butyl]acridine-4-carboxamide and d(CG(5Br)UACG)(2) and d(CGTACG)(2) have been solved by X-ray crystallography using MAD phasing methodology and refined to a resolution of 1.6 A. The complexes crystallised in space group C222. An asymmetric unit in the brominated complex comprises two strands of DNA, one disordered drug molecule, two cobalt (II) ions and 19 water molecules (31 in the native complex). Asymmetric units in the native complex also contain a sodium ion. The structures exhibit novel features not previously observed in crystals of DNA/drug complexes. The DNA helices stack in continuous columns with their central 4 bp adopting a B-like motif. However, despite being a palindromic sequence, the terminal GC base pairs engage in quite different interactions. At one end of the duplex there is a CpG dinucleotide overlap modified by ligand intercalation and terminal cytosine exchange between symmetry-related duplexes. A novel intercalation complex is formed involving four DNA duplexes, four ligand molecules and two pairs of base tetrads. The other end of the DNA is frayed with the terminal guanine lying in the minor groove of the next duplex in the column. The structure is stabilised by guanine N7/cobalt (II) coordination. We discuss our findings with respect to the effects of packing forces on DNA crystal structure, and the potential effects of intercalating agents on biochemical processes involving DNA quadruplexes and strand exchanges. NDB accession numbers: DD0032 (brominated) and DD0033 (native). PMID- 11058125 TI - Synthesis and monitored selection of 5'-nucleobase-capped oligodeoxyribonucleotides. AB - Oligodeoxynucleotides bearing 5'-appendages consisting of a nucleobase and an amide linkage were prepared from 5'-amino-5'-deoxyoligonucleotides, amino acid building blocks and thymine or uracil derivatives. Small chemical libraries of 5' modified oligonucleotides bearing the nucleobase moieties via five, three or two atom linkages were subjected to spectrometrically monitored nuclease selections to identify members with high affinity for target strands. The smallest of the appendages tested, a uracil acetic acid substituent, was found to convey the greatest duplex stabilizing effect on the octamer 5'-T*GGTTGAC-3', where T* denotes the 5'-amino-5'-deoxythymidine residue. Compared to 5'-TTGGTTGAC-3', the modified sequence 5'-u-T*GGTTGAC-3' gives a duplex with 5'-GTCAACCAA-3' that melts 4 degrees C higher. The duplex-stabilizing effect of this 5'-substituent does not require a specific residue at the 3'-terminus of the complement and the available data suggest that the uracil moiety is located in the major groove of the duplex. PMID- 11058126 TI - In vitro selection of an RNA sequence that interacts with high affinity with thymidylate synthase. AB - Previous studies have shown that the repressive effect of thymidylate synthase (TS) mRNA translation is mediated by direct binding of TS itself to two cis acting elements on its cognate mRNA. To identify the optimal RNA nucleotides that interact with TS, we in vitro synthesized a completely degenerate, linear RNA pool of 25 nt and employed in vitro selection to isolate high affinity RNA ligands that bind human TS protein. After 10 rounds of selection and amplification, a single RNA molecule was selected that bound TS protein with nearly 20-fold greater affinity than native, wild-type TS RNA sequences. Secondary structure analysis of this RNA sequence predicted it to possess a stem loop structure. Deletion and/or modification of the UGU loop element within the RNA sequence decreased binding to TS by up to 1000-fold. In vivo transfection experiments revealed that the presence of the selected RNA sequence resulted in a significant increase in the expression of a heterologous luciferase reporter construct in human colon cancer H630 and TS-overexpressing HCT-C:His-TS+ cells, but not in HCT-C18 cells expressing a functionally inactive TS. In addition, the presence of this element in H630 cells leads to induced expression of TS protein. An immunoprecipitation method using RT-PCR confirmed a direct interaction between human TS protein and the selected RNA sequence in transfected human cancer H630 cells. This study identified a novel RNA sequence from a degenerate RNA library that specifically interacts with TS. PMID- 11058127 TI - Molecular characterisation of RecQ homologues in Arabidopsis thaliana. AB - Members of the RecQ family of DNA helicases are involved in processes linked to DNA replication, DNA recombination and gene silencing. RecQ homologues of various animals have been described recently. Here, for the first time for plants, we characterised cDNAs of all in all six different RecQ-like proteins that are expressed to different extents in Arabidopsis thaliana. Surprisingly, three of these proteins are small in size [AtRecQl1, AtRecQl2, AtRecQl3-606, 705 and 713 amino acids (aa), respectively], whereas the two bigger proteins result from a duplication event during plant evolution [AtRecQl4A and AtRecQl4B-1150 and 1182 aa, respectively]. Another homologue (AtRecQsim, 858 aa) most probably arose by insertion of an unrelated sequence within its helicase domain. The presence of these homologues demonstrates the conservation of RecQ family functions in higher eukaryotes. We also detected a small gene (AtWRNexo) encoding 285 aa which, being devoid of any RecQ-like helicase domain, reveals a striking homology to the exonuclease domain of human Werner protein, a prominent RecQ helicase of larger size. By means of the two-hybrid assay we were able to detect an interaction between AtWRNexo and AtRecQl2, indicating that activities that reside in a single protein chain in mammals might in plants be complemented in trans. PMID- 11058128 TI - Blocking transcription of the human rhodopsin gene by triplex-mediated DNA photocrosslinking. AB - To explore the ability of triplex-forming oligodeoxyribonucleotides (TFOs) to inhibit genes responsible for dominant genetic disorders, we used two TFOs to block expression of the human rhodopsin gene, which encodes a G protein-coupled receptor involved in the blinding disorder autosomal dominant retinitis pigmentosa. Psoralen-modified TFOs and UVA irradiation were used to form photoadducts at two target sites in a plasmid expressing a rhodopsin-EGFP fusion, which was then transfected into HT1080 cells. Each TFO reduced rhodopsin-GFP expression by 70-80%, whereas treatment with both reduced expression by 90%. Expression levels of control genes on either the same plasmid or one co transfected were not affected by the treatment. Mutations at one TFO target eliminated its effect on transcription, without diminishing inhibition by the other TFO. Northern blots indicated that TFO-directed psoralen photoadducts blocked progression of RNA polymerase, resulting in truncated transcripts. Inhibition of gene expression was not relieved over a 72 h period, suggesting that TFO-induced psoralen lesions are not repaired on this time scale. Irradiation of cells after transfection with plasmid and psoralen-TFOs produced photoadducts inside the cells and also inhibited expression of rhodopsin-EGFP. We conclude that directing DNA damage with psoralen-TFOs is an efficient and specific means for blocking transcription from the human rhodopsin gene. PMID- 11058129 TI - The transcriptional co-activator P/CAF potentiates TGF-beta/Smad signaling. AB - Smads perform pivotal functions in the intracellular signaling of transforming growth factor-beta (TGF-beta). TGF-beta-mediated activation of TGF-beta type I receptor stimulates the phosphorylation of Smad2 and Smad3 and subsequent heteromeric complex formation with Smad4. The heteromeric Smad complexes translocate into the nucleus where they, in co-operation with co-activators and co-repressors, regulate transcriptional responses. Here we investigated the possible co-activator function of P/CAF in TGF-beta/Smad signaling. P/CAF was found to interact directly with Smad3 in vitro. Moreover, Smad2 and Smad3 interacted with P/CAF upon TGF-beta type I receptor activation in cultured mammalian cells. The interaction involves the MH2 domain of Smad3 and the N terminal region of P/CAF. P/CAF potentiated the transcriptional activity of heterologous Gal4-Smad2 and Gal4-Smad3 fusion proteins. In addition, P/CAF potentiated the TGF-beta/Smad3-induced transcriptional responses, which could be further enhanced by co-activators p300 and Smad4. P/CAF may, therefore, activate Smad-mediated transcriptional responses independently or in co-operation with p300/CBP. Our results indicate a direct physical and functional interplay between two negative regulators of cell proliferation, Smad3 and P/CAF. PMID- 11058130 TI - Archaeal RNA polymerase subunits F and P are bona fide homologs of eukaryotic RPB4 and RPB12. AB - The archaeal and eukaryotic evolutionary domains diverged from each other approximately 2 billion years ago, but many of the core components of their transcriptional and translational machineries still display a readily recognizable degree of similarity in their primary structures. The F and P subunits present in archaeal RNA polymerases were only recently identified in a purified archaeal RNA polymerase preparation and, on the basis of localized sequence homologies, tentatively identified as archaeal versions of the eukaryotic RPB4 and RPB12 RNA polymerase subunits, respectively. We prepared recombinant versions of the F and P subunits from Methanococcus jannaschii and used them in in vitro and in vivo protein interaction assays to demonstrate that they interact with other archaeal subunits in a manner predicted from their eukaryotic counterparts. The overall structural conservation of the M. jannaschii F subunit, although not readily recognizable on the primary amino acid sequence level, is sufficiently high to allow the formation of an archaeal-human F-RPB7 hybrid complex. PMID- 11058131 TI - Characterization of the interaction between alphaCP(2) and the 3'-untranslated region of collagen alpha1(I) mRNA. AB - Activated hepatic stellate cells produce increased type I collagen in hepatic fibrosis. The increase in type I collagen protein results from an increase in mRNA levels that is mainly mediated by increased mRNA stability. Protein-RNA interactions in the 3'-UTR of the collagen alpha1(I) mRNA correlate with stabilization of the mRNA during hepatic stellate cell activation. A component of the binding complex is alphaCP(2). Recombinant alphaCP(2) is sufficient for binding to the 3'-UTR of collagen alpha1(I). To characterize the binding affinity of and specificity for alphaCP(2), we performed electrophoretic mobility shift assays using the poly(C)-rich sequence in the 3'-UTR of collagen alpha1(I) as probe. The binding affinity of alphaCP(2) for the 3'-UTR sequence is approximately 2 nM in vitro and the wild-type 3' sequence binds with high specificity. Furthermore, we demonstrate a system for detecting protein nucleotide interactions that is suitable for high throughput assays using molecular beacons. Molecular beacons, developed for DNA-DNA hybridization, are oligonucleotides with a fluorophore and quencher brought together by a hairpin sequence. Fluorescence increases when the hairpin is disrupted by binding to an antisense sequence or interaction with a protein. Molecular beacons displayed a similar high affinity for binding to recombinant alphaCP(2) to the wild-type 3' sequence, although the kinetics of binding were slower. PMID- 11058132 TI - Complete genome sequence of the alkaliphilic bacterium Bacillus halodurans and genomic sequence comparison with Bacillus subtilis. AB - The 4 202 353 bp genome of the alkaliphilic bacterium Bacillus halodurans C-125 contains 4066 predicted protein coding sequences (CDSs), 2141 (52.7%) of which have functional assignments, 1182 (29%) of which are conserved CDSs with unknown function and 743 (18. 3%) of which have no match to any protein database. Among the total CDSs, 8.8% match sequences of proteins found only in Bacillus subtilis and 66.7% are widely conserved in comparison with the proteins of various organisms, including B.subtilis. The B. halodurans genome contains 112 transposase genes, indicating that transposases have played an important evolutionary role in horizontal gene transfer and also in internal genetic rearrangement in the genome. Strain C-125 lacks some of the necessary genes for competence, such as comS, srfA and rapC, supporting the fact that competence has not been demonstrated experimentally in C-125. There is no paralog of tupA, encoding teichuronopeptide, which contributes to alkaliphily, in the C-125 genome and an ortholog of tupA cannot be found in the B.subtilis genome. Out of 11 sigma factors which belong to the extracytoplasmic function family, 10 are unique to B. halodurans, suggesting that they may have a role in the special mechanism of adaptation to an alkaline environment. PMID- 11058133 TI - The DNA strand of chimeric RNA/DNA oligonucleotides can direct gene repair/conversion activity in mammalian and plant cell-free extracts. AB - Chimeric oligonucleotides (chimeras), consisting of RNA and DNA bases folded by complementarity into a double hairpin conformation, have been shown to alter or repair single bases in plant and animal genomes. An uninterrupted stretch of DNA bases within the chimera is known to be active in the sequence alteration while RNA residues aid in complex stability. In this study, the two strands were separated in the hope of defining the role each plays in conversion. Using a series of single-stranded oligonucleotides, comprised of all RNA or DNA residues and various mixtures, several new structures have emerged as viable molecules in nucleotide conversion. When extracts from mammalian and plant cells and a genetic readout assay in bacteria are used, single-stranded oligonucleotides, containing a defined number of thioate backbone modifications, were found to be more active than the original chimera structure in the process of gene repair. Single stranded oligonucleotides containing fully modified backbones were found to have low repair activity and in fact induce mutation. Molecules containing various lengths of modified RNA bases (2'-O-methyl) were also found to possess low activity. Taken together, these results confirm the directionality of nucleotide conversion by the DNA strand of the chimera and further present a novel, modified single-stranded DNA molecule that directs conversion in plant and animal cell free extracts. PMID- 11058134 TI - Mutant alleles of Schizosaccharomyces pombe rad9(+) alter hydroxyurea resistance, radioresistance and checkpoint control. AB - Schizosaccharomyces pombe rad9 mutations can render cells sensitive to hydroxyurea (HU), gamma-rays and UV light and eliminate associated checkpoint controls. In vitro mutagenesis was performed on S.pombe rad9 and altered alleles were transplaced into the genome to ascertain the functional significance of five groups of evolutionarily conserved amino acids. Most targeted regions were changed to alanines, whereas rad9-S3 encodes a protein devoid of 22 amino acids normally present in yeast but absent from mammalian Rad9 proteins. We examined whether these rad9 alleles confer radiation and HU sensitivity and whether the sensitivities correlate with checkpoint control deficiencies. One rad9 mutant allele was fully active, whereas four others demonstrated partial loss of function. rad9-S1, which contains alterations in a BH3-like domain, conferred HU resistance but increased sensitivity to gamma-rays and UV light, without affecting checkpoint controls. rad9-S2 reduced gamma-ray sensitivity marginally, without altering other phenotypes. Two alleles, rad9-S4 and rad9-S5, reduced HU sensitivity, radiosensitivity and caused aberrant checkpoint function. HU-induced checkpoint control could not be uncoupled from drug resistance. These results establish unique as well as overlapping functional domains within Rad9p and provide evidence that requirements of the protein for promoting resistance to radiation and HU are not identical. PMID- 11058135 TI - The age-related accumulation of a mitochondrial DNA control region mutation in muscle, but not brain, detected by a sensitive PNA-directed PCR clamping based method. AB - The peptide nucleic acid (PNA)-directed PCR clamping technique was modified and applied to the detection of mitochondrial DNA mutations with low heteroplasmy. This method is extremely specific, eliminating false positives in the absence of mutant molecules, and highly sensitive, being capable of detecting mutations at the level of 0.1% of total molecules. Moreover, the reaction can be multiplexed to identify more than one mutation per reaction. Using this technique, the levels of three point mutations, the tRNA(Leu(UUA)) 3243 mutation causing mitochondrial encephalopathy, lactic acidosis and stroke-like episodes (MELAS); the tRNA(Lys) 8344 mutation causing myoclonic epilepsy and ragged red fibers (MERRF); and the nucleotide position 414 mutation adjacent to the control region promoters, were evaluated in human brain and muscle from individuals of various ages. While none of the mutations were detected in brain samples from individuals ranging in age from 23 to 93, the 414 mutation could be detected in muscle from individuals 30 years and older. These data demonstrate that the 3243 and 8344 mutations do not accumulate with age to levels greater than 0.1% in brain and muscle. By contrast, the 414 mutation accumulates with age in normal human muscle, though not in brain. The reason for the striking absence of the 414 mutation in aging brain is unknown. PMID- 11058136 TI - B-form to A-form conversion by a 3'-terminal ribose: crystal structure of the chimera d(CCACTAGTG)r(G). AB - The crystal structure of the chimerical decamer d(CCACTAGTG)r(G), bearing a 3' terminal ribo-guanidine, has been solved and refined at 1.8 A resolution (R factor 16.6%; free R-factor 22.8%). The decamer crystallizes in the orthorhombic space group P2(1)2(1)2(1) with unit cell constants a = 23.90 A, b = 45.76 A and c = 49.27 A. The structure was solved by molecular replacement using the coordinates of the isomorphous chimera r(GCG)d(TATACGC). The final model contains one duplex and 77 water molecules per asymmetric unit. Surprisingly, all residues adopt a conformation typical for A-form nucleic acids (C3'-endo type sugar pucker) although the all-DNA analog, d(CCACTAGTGG), has been crystallized in the B-form. Comparing circular dichroism spectra of the chimera and the corresponding all-DNA sequence reveals a similar trend of the former molecule to adopt an A like conformation in solution. The results suggest that the preference of ribonucleotides for the A-form is communicated into the 5'-direction of an oligonucleotide strand, although direct interactions of the 2'-hydroxyl group can only be discerned with nucleotides in the 3'-direction of a C3'-endo puckered ribose. These observations imply that forces like water-mediated contacts, the concerted motions of backbone torsion angles, and stacking preferences, are responsible for such long-range influences. This bi-directional structural communication originating from a ribonucleotide can be expected to contribute to the stability of the A-form within all-RNA duplexes. PMID- 11058137 TI - Analysis of canonical and non-canonical splice sites in mammalian genomes. AB - A set of 43 337 splice junction pairs was extracted from mammalian GenBank annotated genes. Expressed sequence tag (EST) sequences support 22 489 of them. Of these, 98.71% contain canonical dinucleotides GT and AG for donor and acceptor sites, respectively; 0.56% hold non-canonical GC-AG splice site pairs; and the remaining 0.73% occurs in a lot of small groups (with a maximum size of 0.05%). Studying these groups we observe that many of them contain splicing dinucleotides shifted from the annotated splice junction by one position. After close examination of such cases we present a new classification consisting of only eight observed types of splice site pairs (out of 256 a priori possible combinations). EST alignments allow us to verify the exonic part of the splice sites, but many non-canonical cases may be due to intron sequencing errors. This idea is given substantial support when we compare the sequences of human genes having non-canonical splice sites deposited in GenBank by high throughput genome sequencing projects (HTG). A high proportion (156 out of 171) of the human non canonical and EST-supported splice site sequences had a clear match in the human HTG. They can be classified after corrections as: 79 GC-AG pairs (of which one was an error that corrected to GC-AG), 61 errors that were corrected to GT-AG canonical pairs, six AT-AC pairs (of which two were errors that corrected to AT AC), one case was produced from non-existent intron, seven cases were found in HTG that were deposited to GenBank and finally there were only two cases left of supported non-canonical splice sites. If we assume that approximately the same situation is true for the whole set of annotated mammalian non-canonical splice sites, then the 99.24% of splice site pairs should be GT-AG, 0.69% GC-AG, 0.05% AT-AC and finally only 0.02% could consist of other types of non-canonical splice sites. We analyze several characteristics of EST-verified splice sites and build weight matrices for the major groups, which can be incorporated into gene prediction programs. We also present a set of EST-verified canonical splice sites larger by two orders of magnitude than the current one (22 199 entries versus approximately 600) and finally, a set of 290 EST-supported non-canonical splice sites. Both sets should be significant for future investigations of the splicing mechanism. PMID- 11058138 TI - Synthesis and crystal structure of an octamer RNA r(guguuuac)/r(guaggcac) with G.G/U.U tandem wobble base pairs: comparison with other tandem G.U pairs. AB - We have determined the crystal structure of the RNA octamer duplex r(guguuuac)/r(guaggcac) with a tandem wobble pair, G.G/U.U (motif III), to compare it with U.G/G.U (motif I) and G.U/U.G (motif II) and to better understand their relative stabilities. The crystal belongs to the rhombohedral space group R3. The hexagonal unit cell dimensions are a = b = 41.92 A, c = 56.41 A, and gamma = 120 degrees, with one duplex in the asymmetric unit. The structure was solved by the molecular replacement method at 1.9 A resolution and refined to a final R: factor of 19.9% and R(free) of 23.3% for 2862 reflections in the resolution range 10.0-1.9 A with F >/= 2sigma(F). The final model contains 335 atoms for the RNA duplex and 30 water molecules. The A-RNA stacks in the familiar head-to-tail fashion forming a pseudo-continuous helix. The uridine bases of the tandem U.G pairs have slipped towards the minor groove relative to the guanine bases and the uridine O2 atoms form bifurcated hydrogen bonds with the N1 and N2 of guanines. The N2 of guanine and O2 of uridine do not bridge the 'locked' water molecule in the minor groove, as in motifs I and II, but are bridged by water molecules in the major groove. A comparison of base stacking stabilities of motif III with motifs I and II confirms the result of thermodynamic studies, motif I > motif III > motif II. PMID- 11058139 TI - Two novel dATP analogs for DNA photoaffinity labeling. AB - Two new photoreactive dATP analogs, N(6)-[4-azidobenzoyl-(2-aminoethyl)]-2' deoxyadenosine-5'-triphospha+ ++ te (AB-dATP) and N(6)-[4-[3-(trifluoromethyl) diazirin-3-yl]benzoyl-(2-aminoethyl) ]-2 '-deoxyadenosine-5'-triphosphate (DB dATP), were synthesized from 2'-deoxyadenosine-5'-monophosphate in a six step procedure. Synthesis starts with aminoethylation of dAMP and continues with rearrangement of N(1)-(2-aminoethyl)-2'-deoxyadenosine-5'-monophosphate to N(6) (2-aminoethyl)-2'-deoxyadenosine-5'-monophosphate (N(6)-dAMP). Next, N(6)-dAMP is converted into the triphosphate form by first protecting the N-6 primary amino group before coupling the pyrophosphate. After pyrophosphorylation, the material is deprotected to yield N(6)-(2-aminoethyl)-2'-deoxyadenosine-5'-triphosphate (N(6)-dATP). The N-6 amino group is subsequently used to attach either a phenylazide or phenyldiazirine and the photoreactive nucleotide is then enzymatically incorporated into DNA. N(6)-dATP and its photoreactive analogs AB dATP and DB-dATP were successfully incorporated into DNA using the exonuclease free Klenow fragment of DNA polymerase I in a primer extension reaction. UV irradiation of the primer extension reaction with AB-dATP or DB-dATP showed specific photocrosslinking of DNA polymerase I to DNA. PMID- 11058140 TI - The thy pol-2 intein of Thermococcus hydrothermalis is an isoschizomer of PI-TliI and PI-TfuII endonucleases. AB - THY Pol-2 intein, from Thermococcus hydrothermalis, belongs to the same allelic family as TLI Pol-2 (PI-TLII), Tfu Pol-2 (PI-TFUII) and TspTY Pol-3 mini-intein, all inserted at the pol-c site of archaeal DNA polymerase genes. This new intein was cloned, expressed in Escherichia coli and purified. The intein is a specific endonuclease (PI-THY:I) which cleaves the inteinless sequence of the THY DNA pol gene. Moreover, PI-TLII, PI-TFUII and PI-THYI are very similar endonucleases which cleave DNA in the same optimal conditions at 70 degrees C yielding similar 3'-hydroxyl overhangs of 4 bp and the reaction is subject to product inhibition. The three enzymes are able to cleave the three DNA sequences spanning the pol-c site and a 24 bp consensus cleavage site was defined for the three isoschizomers. However, the exact size of the minimal cleavage site depends both on the substrate sequence and the endonuclease. The inability of the isoschizomers to cleave the inteinless DNA polymerase gene from Pyrococcus spp. KOD is due to point substitutions on the 5' side of the pol-c site, suggesting that the absence of inteins of this allelic family in DNA polymerase genes from Pyrococcus spp. can be linked to small differences in the target site sequence. PMID- 11058141 TI - RecA-independent ectopic transposition in vivo of a bacterial group II intron. AB - RmInt1 is a group II intron of Sinorhizobium meliloti which was initially found within the insertion sequence ISRm2011-2. Although the RmInt1 intron-encoded protein lacks a recognizable endonuclease domain, it is able to mediate insertion of RmInt1 at an intron-specific location in intronless ISRm2011-2 recipient DNA, a phenomenon termed homing. Here we have characterized three additional insertion sites of RmInt1 in the genome of S.meliloti. Two of these sites are within IS elements closely related to ISRm2011-2, which appear to form a characteristic group within the IS630-Tc1 family. The third site is in the oxi1 gene, which encodes a putative oxide reductase. The newly identified integration sites contain conserved intron-binding site (IBS1 and IBS2) and delta' sequences (14 bp). The RNA of the intron-containing oxi1 gene is able to splice and the oxi1 site is a DNA target for RmInt1 transposition in vivo. Ectopic transposition of RmInt1 into the oxi1 gene occurs at 20-fold lower efficiency than into the homing site (ISRm2011-2) and is independent of the major RecA recombination pathway. The possibility that transposition of RmInt1 to the oxi1 site occurs by reverse splicing into DNA is discussed. PMID- 11058142 TI - Cre-mediated germline mosaicism: a method allowing rapid generation of several alleles of a target gene. AB - Conditional gene targeting uses the insertion of expression cassettes for the selection of targeted embryonic stem cells. The presence of these cassettes in the final targeted chromosomal locus may affect the normal expression of the targeted gene and produce interesting knock down phenotypes. We show here that the selection cassette may then be selectively removed in vivo, using three appropriately positioned loxP sites in the targeted gene and the transgenic mouse EIIaCre. This strategy was applied to two different target genes and we demonstrated that it is reliable and reproducible. First, we generated double transgenic EIIaCre/loxP mice (F1) that showed variable degrees of mosaicism for partially CRE-recombined floxed alleles. Efficiency of EIIaCre at creating mosaicism was dependent on the target gene and on parental transmission of the transgene. The segregation of partially recombined alleles and EIIaCre transgene was obtained in the next generation using mosaic F1 males. Mosaic females were unsuitable for this purpose because they systematically generated complete excisions during oogenesis. Our strategy is applicable to other approaches based on three loxP sites. As this procedure allows generation of knock down (presence of neo), knockout (total exision of the loxP-flanked sequences) and floxed substrains (excision of the selection cassette) from a single, targeted germline mutation and in a single experiment, its use may become more widespread in conditional mutagenesis. PMID- 11058143 TI - Interactions of Escherichia coli RNA with bacteriophage MS2 coat protein: genomic SELEX. AB - Genomic SELEX is a method for studying the network of nucleic acid-protein interactions within any organism. Here we report the discovery of several interesting and potentially biologically important interactions using genomic SELEX. We have found that bacteriophage MS2 coat protein binds several Escherichia coli mRNA fragments more tightly than it binds the natural, well studied, phage mRNA site. MS2 coat protein binds mRNA fragments from rffG (involved in formation of lipopolysaccharide in the bacterial outer membrane), ebgR (lactose utilization repressor), as well as from several other genes. Genomic SELEX may yield experimentally induced artifacts, such as molecules in which the fixed sequences participate in binding. We describe several methods (annealing of oligonucleotides complementary to fixed sequences or switching fixed sequences) to eliminate some, or almost all, of these artifacts. Such methods may be useful tools for both randomized sequence SELEX and genomic SELEX. PMID- 11058144 TI - PCR hot start using primers with the structure of molecular beacons (hairpin-like structure). AB - A new technique of PCR hot start using oligonucleotide primers with a stem-loop structure is developed here. The molecular beacon oligonucleotide structure without any chromophore addition to the ends was used. The 3'-end sequence of the primers was complementary to the target and five or six nucleotides complementary to the 3'-end were added to the 5'-end. During preparation of the reaction mixture and initial heating, the oligonucleotide has a stem-loop structure and cannot serve as an effective primer for DNA polymerase. After heating to the annealing temperature it acquires a linear structure and primer extension can begin. PMID- 11058145 TI - Efficient gene targeted random mutagenesis in genetically stable Escherichia coli strains. AB - We describe a method to generate in vivo collections of mutants orders of magnitude larger than previously possible. The method favors accumulation of mutations in the target gene, rather than in the host chromosome. This is achieved by propagating the target gene on a plasmid, in Escherichia coli cells, within the region preferentially replicated by DNA polymerase I (Pol I), which replicates only a minor fraction of the chromosome. Mutagenesis is enhanced by a conjunction of a Pol I variant that has a low replication fidelity and the absence of the mutHLS system that corrects replication errors. The method was tested with two reporter genes, encoding lactose repressor or lipase. The proportion of mutants in the collection was estimated to reach 1% after one cycle of growth and 10% upon prolonged cell cultivation, resulting in collections of 10(12)-10(13) mutants per liter of cell culture. The extended cultivation did not affect growth properties of the cells. We suggest that our method is well suited for generating protein variants too rare to be present in the collections established by methods used previously and for isolating the genes that encode such variants by submitting the cells of the collections to appropriate selection protocols. PMID- 11058146 TI - Two tandem verprolin homology domains are necessary for a strong activation of Arp2/3 complex-induced actin polymerization and induction of microspike formation by N-WASP. AB - All WASP family proteins share a common C terminus that consists of the verprolin homology domain (V), cofilin homology domain (C), and acidic region (A), through which they activate Arp2/3 complex-induced actin polymerization. In this study, we characterized the Arp2/3 complex-mediated actin polymerization activity of VCA fragments of all of the WASP family proteins: WASP, N-WASP, WAVE1, WAVE2, and WAVE3. All of the VCA fragments stimulated the nucleating activity of Arp2/3 complex. Among them, N-WASP VCA, which possesses two tandem V motifs, had a more potent activity than other VCA proteins. The chimeric protein experiments revealed that the V motif was more important to the activation potency than the CA region; two V motifs were required for full activity of N-WASP. COS7 cells overexpressing N-WASP form microspikes in response to epidermal growth factor. However, when a chimeric protein in which the VCA region of N-WASP is replaced with WAVE1 VCA was overexpressed, microspike formation was suppressed. Interestingly, when the N-WASP VCA region was replaced with WAVE1 VCA, having two V motifs, this chimeric protein could induce microspike formation. These results indicate that strong activation of Arp2/3 complex by N-WASP is mainly caused by its two tandem V motifs, which are essential for actin microspike formation. PMID- 11058147 TI - Sparing of neuronal function postseizure with gene therapy. AB - Numerous studies have demonstrated that gene therapy interventions can protect neurons from death after neurological insults. In nearly all such studies, however, "protection" consists of reduced neurotoxicity, with no demonstrated preservation of neuronal function. We used a herpes simplex virus-1 system to overexpress either the Glut-1 glucose transporter (GT) (to buffer energetics), or the apoptosis inhibitor Bcl-2. Both decreased hippocampal neuron loss to similar extents during excitotoxic insults in vitro and in vivo. However, the mediating mechanisms and consequences of the two interventions differed. GT overexpression attenuated early, energy-dependent facets of cell death, blocking oxygen radical accumulation. Bcl-2 expression, in contrast, blocked components of death downstream from the energetic and oxidative facets. Most importantly, GT- but not Bcl-2-mediated protection preserved hippocampal function as assessed spatial maze performance. Thus, gene therapeutic sparing of neurons from insult-induced death does not necessarily translate into sparing of function. PMID- 11058148 TI - An empirical explanation of color contrast. AB - For reasons not well understood, the color of a surface can appear quite different when placed in different chromatic surrounds. Here we explore the possibility that these color contrast effects are generated according to what the same or similar stimuli have turned out to signify in the past about the physical relationships between reflectance, illumination, and the spectral returns they produce. This hypothesis was evaluated by (i) comparing the physical relationships of reflectances, illuminants, and spectral returns with the perceptual phenomenology of color contrast and (ii) testing whether perceptions of color contrast are predictably changed by altering the probabilities of the possible sources of the stimulus. The results we describe are consistent with a wholly empirical explanation of color contrast effects. PMID- 11058149 TI - Polyglutamine disease and neuronal cell death. PMID- 11058150 TI - Life in extreme environments: hydrothermal vents. PMID- 11058151 TI - Postsegregational killing does not increase plasmid stability but acts to mediate the exclusion of competing plasmids. AB - Postsegregational killing (PSK) systems consist of a tightly linked toxin antitoxin pair. Antitoxin must be continually produced to prevent the longer lived toxin from killing the cell. PSK systems on plasmids are widely believed to benefit the plasmid by ensuring its stable vertical inheritance. However, experimental tests of this "stability" hypothesis were not consistent with its predictions. We suggest an alternative hypothesis to explain the evolution of PSK: that PSK systems have been selected through benefiting host plasmids in environments where plasmids must compete during horizontal reproduction. In this "competition" hypothesis, success of PSK systems is a consequence of plasmid plasmid competition, rather than from an adaptive plasmid-host relationship. In support of this hypothesis, a plasmid-encoded parDE PSK system mediated the exclusion of an isogenic DeltaparDE plasmid. An understanding of how PSK systems influence plasmid success may provide insight into the evolution of other determinants (e.g., antibiotic resistance and virulence) also rendering a cell potentially dependent on an otherwise dispensable plasmid. PMID- 11058152 TI - Increased resistance to acetaminophen hepatotoxicity in mice lacking glutathione S-transferase Pi. AB - Overdose of acetaminophen, a widely used analgesic drug, can result in severe hepatotoxicity and is often fatal. This toxic reaction is associated with metabolic activation by the P450 system to form a quinoneimine metabolite, N acetyl-p-benzoquinoneimine (NAPQI), which covalently binds to proteins and other macromolecules to cause cellular damage. At low doses, NAPQI is efficiently detoxified, principally by conjugation with glutathione, a reaction catalyzed in part by the glutathione S-transferases (GST), such as GST Pi. To assess the role of GST in acetaminophen hepatotoxicity, we examined acetaminophen metabolism and liver damage in mice nulled for GstP (GstP1/P2((-/-))). Contrary to our expectations, instead of being more sensitive, GstP null mice were highly resistant to the hepatotoxic effects of this compound. No significant differences between wild-type (GstP1/P2((+/+))) mice and GstP1/P2((-/-)) nulls in either the rate or route of metabolism, particularly to glutathione conjugates, or in the levels of covalent binding of acetaminophen-reactive metabolites to cellular protein were observed. However, although a similar rapid depletion of hepatic reduced glutathione (GSH) was found in both GstP1/P2((+/+)) and GstP1/P2((-/-)) mice, GSH levels only recovered in the GstP1/P2((-/-)) mice. These data demonstrate that GstP does not contribute in vivo to the formation of glutathione conjugates of acetaminophen but plays a novel and unexpected role in the toxicity of this compound. This study identifies new ways in which GST can modulate cellular sensitivity to toxic effects and suggests that the level of GST Pi may be an important and contributing factor in the sensitivity of patients with acetaminophen-induced hepatotoxicity. PMID- 11058153 TI - Zidovudine-didanosine coexposure potentiates DNA incorporation of zidovudine and mutagenesis in human cells. AB - Drug combinations that include nucleoside reverse transcriptase inhibitors (NRTIs) are remarkably effective in preventing maternal-viral transmission of HIV during pregnancy. However, there may be potential long-term risks for children exposed in utero. Examination of the genotoxic and mutagenic effects of two NRTIs, zidovudine [AZT (3'-azido-3'-deoxythymidine)] and didanosine [ddI (2',3' dideoxyinosine)], in cultured human lymphoblastoid cells revealed multiplicative synergistic enhancement of AZT-DNA incorporation and mutant frequency induction in response to the combined drug exposure, as compared with single-drug exposures. Dose-related increases in DNA incorporation of AZT (as measured by a competitive RIA) and mutagenicity at the HPRT and TK loci (as assessed by cell cloning assays) were observed in cells exposed in culture to AZT, or equimolar combinations of AZT + ddI, at exposure concentrations ranging from 3 to 30 times the maximum plasma levels found in humans. Because mutagenesis is strongly associated with tumor induction in experimental models, children exposed transplacentally to combinations of NRTIs may be at risk for cancer development later in life. PMID- 11058154 TI - Molecular identification by "suicide PCR" of Yersinia pestis as the agent of medieval black death. AB - Medieval Black Death is believed to have killed up to one-third of the Western European population during the 14th century. It was identified as plague at this time, but recently the causative organism was debated because no definitive evidence has been obtained to confirm the role of Yersinia pestis as the agent of plague. We obtained the teeth of a child and two adults from a 14th century grave in France, disrupted them to obtain the pulp, and applied the new "suicide PCR" protocol in which the primers are used only once. There were no positive controls: Neither Yersinia nor Yersinia DNA were introduced in the laboratory. A negative result is followed by a new test using other primers; a positive result is followed by sequencing. The second and third primer pair used, coding for a part of the pla gene, generated amplicons whose sequence confirmed that it was Y. pestis in 1 tooth from the child and 19/19 teeth from the adults. Negative controls were negative. Attempts to detect the putative alternative etiologic agents Bacillus anthracis and Rickettsia prowazekii failed. Suicide PCR avoids any risk of contamination as it uses a single-shot primer-its specificity is absolute. We believe that we can end the controversy: Medieval Black Death was plague. PMID- 11058156 TI - Advances in random matrix theory, zeta functions, and sphere packing. AB - Over four hundred years ago, Sir Walter Raleigh asked his mathematical assistant to find formulas for the number of cannonballs in regularly stacked piles. These investigations aroused the curiosity of the astronomer Johannes Kepler and led to a problem that has gone centuries without a solution: why is the familiar cannonball stack the most efficient arrangement possible? Here we discuss the solution that Hales found in 1998. Almost every part of the 282-page proof relies on long computer verifications. Random matrix theory was developed by physicists to describe the spectra of complex nuclei. In particular, the statistical fluctuations of the eigenvalues ("the energy levels") follow certain universal laws based on symmetry types. We describe these and then discuss the remarkable appearance of these laws for zeros of the Riemann zeta function (which is the generating function for prime numbers and is the last special function from the last century that is not understood today.) Explaining this phenomenon is a central problem. These topics are distinct, so we present them separately with their own introductory remarks. PMID- 11058155 TI - Behavioral insensitivity to restraint stress, absent fear suppression of behavior and impaired spatial learning in transgenic rats with hippocampal neuropeptide Y overexpression. AB - Exogenous neuropeptide Y (NPY) reduces experimental anxiety in a wide range of animal models. The generation of an NPY-transgenic rat has provided a unique model to examine the role of endogenous NPY in control of stress and anxiety related behaviors using paradigms previously used by pharmacological studies. Locomotor activity and baseline behavior on the elevated plus maze were normal in transgenic subjects. Two robust phenotypic traits were observed. (i) Transgenic subjects showed a markedly attenuated sensitivity to behavioral consequences of stress, in that they were insensitive to the normal anxiogenic-like effect of restraint stress on the elevated plus maze and displayed absent fear suppression of behavior in a punished drinking test. (ii) A selective impairment of spatial memory acquisition was found in the Morris water maze. Control experiments suggest these traits to be independent. These phenotypic traits were accompanied by an overexpression of prepro-NPY mRNA and NPY peptide and decreased NPY-Y1 binding within the hippocampus, a brain structure implicated both in memory processing and stress responses. Data obtained using this unique model support and extend a previously postulated anti-stress action of NPY and provide novel evidence for a role of NPY in learning and memory. PMID- 11058157 TI - Self-organizing biochemical cycles. AB - I examine the plausibility of theories that postulate the development of complex chemical organization without requiring the replication of genetic polymers such as RNA. One conclusion is that theories that involve the organization of complex, small-molecule metabolic cycles such as the reductive citric acid cycle on mineral surfaces make unreasonable assumptions about the catalytic properties of minerals and the ability of minerals to organize sequences of disparate reactions. Another conclusion is that data in the Beilstein Handbook of Organic Chemistry that have been claimed to support the hypothesis that the reductive citric acid cycle originated as a self-organized cycle can more plausibly be interpreted in a different way. PMID- 11058158 TI - Neuronal death enhanced by N-methyl-D-aspartate antagonists. AB - Glutamate promotes neuronal survival during brain development and destroys neurons after injuries in the mature brain. Glutamate antagonists are in human clinical trials aiming to demonstrate limitation of neuronal injury after head trauma, which consists of both rapid and slowly progressing neurodegeneration. Furthermore, glutamate antagonists are considered for neuroprotection in chronic neurodegenerative disorders with slowly progressing cell death only. Therefore, humans suffering from Huntington's disease, characterized by slowly progressing neurodegeneration of the basal ganglia, are subjected to trials with glutamate antagonists. Here we demonstrate that progressive neurodegeneration in the basal ganglia induced by the mitochondrial toxin 3-nitropropionate or in the hippocampus by traumatic brain injury is enhanced by N-methyl-d-aspartate antagonists but ameliorated by alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate antagonists. These observations reveal that N-methyl-d-aspartate antagonists may increase neurodestruction in mature brain undergoing slowly progressing neurodegeneration, whereas blockade of the action of glutamate at alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate receptors may be neuroprotective. PMID- 11058159 TI - Prime type III factors. AB - It is shown that for each 0 < lambda < 1, the free Araki-Woods factor of type III(lambda) cannot be written as a tensor product of two diffuse von Neumann algebras (i.e., is prime) and does not contain a Cartan subalgebra. PMID- 11058160 TI - An essential amino acid induces epithelial beta -defensin expression. AB - Antimicrobial peptides constitute an important component of the mammalian innate immune response. Several types of antimicrobial peptides, including the beta defensins, are produced at epithelial surfaces in response to infectious threats. Here we show that a class of small molecules, including l-isoleucine and several of its analogs, can specifically induce epithelial beta-defensin expression. This induction is transcriptional in nature and involves activation of the NF kappaB/rel family of trans-activating factors. We hypothesize that these substances represent unique markers for the presence of pathogens and are recognized by innate immune pattern recognition receptors. Isoleucine or its analogs ultimately may have clinical utility as novel immunostimulants that could bolster the barrier defenses of mucosal surfaces. PMID- 11058161 TI - RaSH, a rapid subtraction hybridization approach for identifying and cloning differentially expressed genes. AB - Human melanoma cells growth-arrest irreversibly and terminally differentiate on treatment with a combination of fibroblast interferon and the protein kinase C activator mezerein. This experimental protocol also results in a loss of tumorigenic potential and profound changes in gene expression. Various cloning and cDNA microarray strategies are being used to determine the complete spectrum of gene expression changes underlying these alterations in human melanoma cells. An efficient approach, Rapid Subtraction Hybridization (RaSH), has been developed that is permitting the identification of genes of potential relevance to cancer growth control and terminal cell differentiation. RaSH cDNA libraries are prepared from double-stranded cDNAs that are enzymatically digested into small fragments, ligated to adapters, and PCR amplified followed by incubation of tester and driver PCR fragments. This subtraction hybridization scheme is technically simple and results in the identification of a high proportion of differentially expressed sequences, including known genes and those not described in current DNA databases. The RaSH approach represents an efficient methodology for identifying and cloning genes displaying differential expression that associate with and potentially regulate complex biological processes. PMID- 11058162 TI - Sensory deprivation without competition yields modest alterations of short-term synaptic dynamics. AB - Cortical maps express experience-dependent plasticity. However, the underlying cellular mechanisms remain unclear. We have recently shown that sensory deprivation results in large changes of the short-term dynamics of excitatory synapses at the junction of deprived and spared somatosensory (barrel) cortex, which may contribute to map reorganization. A key issue is whether the alterations in short-term synaptic dynamics are driven by a loss of sensory input or by competition between deprived and spared inputs. Here, we report that short term dynamics of horizontal pathways in the middle of uniformly deprived cortex change only modestly. Vertical intracortical pathways were unaffected by deprivation. Our results suggest that uniform loss of sensory activity has a limited effect on short-term synaptic dynamics. We concluded that competition between sensory inputs is necessary to produce large-scale changes in synaptic dynamics after sensory deprivation. PMID- 11058163 TI - RNA polymerase III transcription factor TFIIIC2 is overexpressed in ovarian tumors. AB - Most transformed cells display abnormally high levels of RNA polymerase (pol) III transcripts. Although the full significance of this is unclear, it may be fundamental because healthy cells use two key tumor suppressors to restrain pol III activity. We present the first evidence that a pol III transcription factor is overexpressed in tumors. This factor, TFIIIC2, is a histone acetyltransferase that is required for synthesis of most pol III products, including tRNA and 5S rRNA. TFIIIC2 is a complex of five polypeptides, and mRNAs encoding each of these subunits are overexpressed in human ovarian carcinomas; this may explain the elevated TFIIIC2 activity that is found consistently in the tumors. Deregulation in these cancers is unlikely to be a secondary response to rapid proliferation, because there is little or no change in TFIIIC2 mRNA levels when actively cycling cells are compared with growth-arrested cells in culture. Using purified factors, we show that raising the level of TFIIIC2 is sufficient to stimulate pol III transcription in ovarian cell extracts. The data suggest that overexpression of TFIIIC2 contributes to the abnormal abundance of pol III transcripts in ovarian tumors. PMID- 11058164 TI - LEUNIG, a putative transcriptional corepressor that regulates AGAMOUS expression during flower development. AB - Regulation of homeotic gene expression is critical for proper developmental patterns in both animals and plants. LEUNIG is a key regulator of the Arabidopsis floral homeotic gene AGAMOUS. Mutations in LEUNIG cause ectopic AGAMOUS mRNA expression in the outer two whorls of a flower, leading to homeotic transformations of floral organ identity as well as loss of floral organs. We isolated the LEUNIG gene by using a map-based approach and showed that LEUNIG encodes a glutamine-rich protein with seven WD repeats and is similar in motif structure to a class of functionally related transcriptional corepressors including Tup1 from yeast and Groucho from Drosophila. The nuclear localization of LEUNIG-GFP is consistent with a role of LEUNIG as a transcriptional regulator. The detection of LEUNIG mRNA in all floral whorls at the time of their inception suggests that the restricted activity of LEUNIG in the outer two floral whorls must depend on interactions with other spatially restricted factors or on posttranslational regulation. Our finding suggests that both animals and plants use similar repressor proteins to regulate critical developmental processes. PMID- 11058165 TI - Does the granular matter? AB - Granular materials, such as sand, gravel, powders, and pharmaceutical pills, are large aggregates of macroscopic, individually solid particles, or "grains." Far from being simple materials with simple properties, they display an astounding range of complex behavior that defies their categorization as solid, liquid, or gas. Just consider how sand can stream through the orifice of an hourglass yet support one's weight on the beach; how it can form patterns strikingly similar to a liquid when vibrated, yet respond to stirring by "unmixing" of large and small grains. Despite much effort, there still is no comprehensive understanding of other forms of matter, like ordinary fluids or solids. In what way, therefore, is granular matter special, and what makes it so difficult to understand? An emerging interdisciplinary approach to answering these questions focuses directly on the material's discontinuous granular nature. PMID- 11058166 TI - Potentiation of pathogen-specific defense mechanisms in Arabidopsis by beta aminobutyric acid. AB - The nonprotein amino acids gamma-aminobutyric acid (GABA) and beta-aminobutyric acid (BABA) have known biological effects in animals and plants. Their mode of action has been the object of thorough research in animals but remains unclear in plants. Our objective was to study the mode of action of BABA in the protection of Arabidopis plants against virulent pathogens. BABA protected Arabidopsis against the oomycete pathogen Peronospora parasitica through activation of natural defense mechanisms of the plant such as callose deposition, the hypersensitive response, and the formation of trailing necroses. BABA was still fully protective against P. parasitica in transgenic plants or mutants impaired in the salicylic acid, jasmonic acid, and ethylene signaling pathways. Treatment with BABA did not induce the accumulation of mRNA of the systemic acquired resistance (SAR)-associated PR-1 and the ethylene- and jasmonic acid-dependent PDF1.2 genes. However, BABA potentiated the accumulation of PR-1 mRNA after attack by virulent pathogenic bacteria. As a result, BABA-treated Arabidopsis plants were less diseased compared with the untreated control. In the case of bacteria, BABA protected mutants insensitive to jasmonic acid and ethylene but was not active in plants impaired in the SAR transduction pathway. Thus, BABA protects Arabidopsis against different virulent pathogens by potentiating pathogen-specific plant resistance mechanisms. In addition, we provide evidence that BABA-mediated papilla formation after P. parasitica infection is independent of the SAR signaling pathway. PMID- 11058167 TI - Selective requirement for c-Rel during IL-12 P40 gene induction in macrophages. AB - A major challenge in the study of gene regulation by NF-kappaB/Rel transcription factors is to understand, at the biological and mechanistic levels, the selective functions of individual Rel family members. To study selectivity, we have examined the NF-kappaB/Rel protein binding site (Rel site) within the IL-12 p40 promoter. IL-12 is a proinflammatory cytokine expressed by activated macrophages that serves as an essential inducer of T helper 1 cell development. In nuclear extracts from lipopolysaccharideactivated macrophages, the predominant Rel dimers capable of binding the IL-12 p40 Rel site were the p50/p65 and p50/c-Rel heterodimers and p50/p50 homodimer. The two heterodimers bound the site with comparable affinities and exhibited comparable transactivation activities. In striking contrast, p40 mRNA and protein concentrations were reduced dramatically in c-Rel(-/-) macrophages and only modestly in p65(-/-) macrophages. Other proinflammatory cytokine mRNAs and proteins were not significantly reduced in c Rel(-/-) macrophages. These results reveal that a c-Rel-containing complex is an essential and selective activator of p40 transcription, which may reflect unique regulatory mechanisms or biological functions of IL-12. Furthermore, because selectivity was not observed in vitro or in transient transactivation experiments, these findings suggest that an understanding of the selectivity mechanism may require an analysis of the endogenous p40 locus. PMID- 11058168 TI - Single 8-oxo-guanine and 8-oxo-adenine lesions induce marked changes in the backbone structure of a 25-base DNA strand. AB - Structural changes in a 25-base DNA strand, induced by single 8-oxo-guanine or 8 oxo-adenine substitutions, were shown by using Fourier transform-infrared spectroscopy with multivariate statistics. Pronounced differences were demonstrated between the parent and derivatives with respect to base interactions and changes in the phospho-deoxyribose backbone. The greatest degree of change in the backbone likely occurred immediately adjacent to the 8-oxo group, potentially altering the stereochemistry at a distance. The 8-oxo lesions, formed from reactive oxygen species (e.g., hydroxyl radicals), may appreciably alter the conformational properties of strands at the replication fork, thus affecting the selectivity of polymerases, the proofreading capability of repair enzymes, and the fidelity of the transcriptional machinery. PMID- 11058169 TI - Chimeras of Dictyostelium myosin II head and neck domains with Acanthamoeba or chicken smooth muscle myosin II tail domain have greatly increased and unregulated actin-dependent MgATPase activity. AB - Phosphorylation of the regulatory light chain of Dictyostelium myosin II increases V(max) of its actin-dependent MgATPase activity about 5-fold under normal assay conditions. Under these assay conditions, unphosphorylated chimeric myosins in which the tail domain of the Dictyostelium myosin II heavy chain is replaced by either the tail domain of chicken gizzard smooth muscle or Acanthamoeba myosin II are 20 times more active because of a 10- to 15-fold increase in V(max) and a 2- to 7-fold decrease in apparent K(ATPase) and are only slightly activated by regulatory light chain phosphorylation. Actin-dependent MgATPase activity of the Dictyostelium/Acanthamoeba chimera is not affected by phosphorylation of serine residues in the tail whose phosphorylation completely inactivates wild-type Acanthamoeba myosin II. These results indicate that the actin-dependent MgATPase activity of these myosins involves specific, tightly coupled, interactions between head and tail domains. PMID- 11058170 TI - Coevolution of head, neck, and tail domains of myosin heavy chains. AB - Myosins, a large family of actin-based motors, have one or two heavy chains with one or more light chains associated with each heavy chain. The heavy chains have a (generally) N-terminal head domain with an ATPase and actin-binding site, followed by a neck domain to which the light chains bind, and a C-terminal tail domain through which the heavy chains self-associate and/or bind the myosin to its cargo. Approximately 140 members of the myosin superfamily have been grouped into 17 classes based on the sequences of their head domains. I now show that a phylogenetic tree based on the sequences of the combined neck and tail domains groups 144 myosins, with a few exceptions, into the same 17 classes. For the nine myosin classes that have multiple members, phylogenetic trees based on the head domain or the combined neck/tail domains are either identical or very similar. For class II myosins, very similar phylogenetic trees are obtained for the head, neck, and tail domains of 47 heavy chains and for 29 essential light chains and 19 regulatory light chains. These data strongly suggest that the head, neck, and tail domains of all myosin heavy chains, and light chains at least of class II myosins, have coevolved and are likely to be functionally interdependent, consistent with biochemical evidence showing that regulated actin-dependent MgATPase activity of Dictyostelium myosin II requires isoform specific interactions between the heavy chain head and tail and light chains. PMID- 11058171 TI - Introduction to frontiers of science. PMID- 11058172 TI - Anti-inflammatories for cardiovascular disease. PMID- 11058173 TI - Natural selection and speciation. PMID- 11058174 TI - Pausing to decide. PMID- 11058175 TI - Blaming the wrong villain. PMID- 11058177 TI - The Michigan Supported Education Program. AB - With the advent of improved pharmacological treatments, empirically tested psychiatric rehabilitation techniques, and an increased emphasis on the empowerment of mental health consumers, many adults with psychiatric disabilities now have a realistic chance of reentering their communities and reestablishing meaningful and productive lives. Because work is a fundamental component of adjustment in adult life, helping individuals obtain and maintain jobs has been viewed as the sine qua non of psychiatric rehabilitation. More recently, however, rehabilitation practitioners have realized that many adults with psychiatric disabilities have the desire and the requisite motivation and educational background to attend college (1). Hence rehabilitation practitioners have recognized that helping individuals restart their postsecondary educational pursuits is a desirable, valid, and viable option (2,3). Supported education is being used increasingly to encourage adults with mental illness to enroll in and complete postsecondary education by providing assistance, preparation, and ongoing counseling (4). Several reports have suggested that supported education programs contribute to positive outcomes such as graduation, acquisition of marketable skills, employment, and positive self-esteem (5,6,7). In this month's column, Carol Mowbray, Ph.D., describes the Michigan Supported Education Program and provides a rationale and empirical validation for its inclusion as an integral modality of psychiatric rehabilitation. PMID- 11058178 TI - Mental health parity and employer-sponsored health insurance in 1999-2000: I. limits. PMID- 11058179 TI - Using case vignettes to train clinicians and utilization reviewers to make level of-care decisions. AB - Dr. Rosenquist and his colleagues describe how their academically based health maintenance organization joined in training for level-of-care decision making with the external managed behavioral health organization that was providing utilization review and case management decisions. The academic department later took over its own utilization review and in so doing internalized the utilization review function. This development, which is beginning to occur in several states, is an important solution to the "assault" that many providers of care have experienced as a result of the utilization review process. Having taken this step to deal with the realities of 21st-century health care, the authors then seize the opportunity to use their own data to improve decision making within the clinic. This process is how we get to best practices. PMID- 11058180 TI - Disability law and the administration of psychotropic medication in the school setting. PMID- 11058181 TI - The impact of terminating disability benefits for substance abusers on substance use and treatment participation. PMID- 11058182 TI - The role of psychotherapy in public psychiatry today. PMID- 11058183 TI - Web sites worth watching. PMID- 11058184 TI - The effects of public managed care on patterns of intensive use of inpatient psychiatric services. PMID- 11058185 TI - Managed care and health care reform: comedy, tragedy, and lessons. PMID- 11058186 TI - Excluding institutions for mental diseases from federal reimbursementfor services: strategy or tragedy? AB - A key component in the dehospitalization of persons with chronic mental illnesses and their translocation to a wide range of settings has been the shift from state to federal funding encouraged by the Social Security Administration's restrictions on funding for institutions for mental diseases (IMDs), usually referred to as the IMD exclusion. The overall effect of the exclusion, which limits federal funding for mentally ill patients receiving care in many settings, including state psychiatric hospitals, has been to create incentives for states to move patients out of state hospitals, which has contributed to homelessness and inappropriate incarceration. The author traces the background and development of the IMD exclusion, starting with the federal government's actions in the 1840s ensuring that the states, not the federal government, would continue to fund care for their seriously mentally ill citizens. He also analyzes Congress' many missed opportunities to pass legislation that would have modified, or perhaps even made moot, the IMD exclusion, including the Clinton administration's efforts at health reform, the quest for parity of insurance coverage for mental illnesses, and the initiation of public-sector managed care. The consequences of the continuation of the IMD exclusion are explored, and the intended fiscal consequences are contrasted with the unintended clinical outcomes. PMID- 11058187 TI - The changing role of dynamic psychotherapy in psychiatric practice. AB - The author reviews the range of accepted indications for dynamic psychotherapy when he first began practice after World War II and describes factors that have played a role in the current undervaluing of this treatment approach. He attributes much of the change to research that has produced a different understanding of many of the conditions treated by psychiatrists and has placed greater emphasis on their medical and biological aspects than on their psychological aspects. He also attributes many alterations in current practice to the change from a two-party to a three-party reimbursement system for psychiatric services. On the basis of his practice, the author illustrates his belief that dynamic therapy continues to have a role in today's psychiatric practice. He describes one category of patients in particular-those who seek treatment for "problems of living"-who can be helped by dynamic psychotherapy. The author makes a case for freeing dynamic therapy from the need to rely exclusively on the criteria of the medical model for its legitimacy. PMID- 11058188 TI - Psychodynamic approaches to the patient. PMID- 11058189 TI - A meta-analysis of the effectiveness of mental health case management over 20 years. AB - OBJECTIVE: Meta-analytical methods were used to investigate the effectiveness of case management and to compare outcomes for assertive community treatment and clinical case management. METHODS: Controlled studies of case management published between 1980 and 1998 were identified from reviews and through database searches. The results were quantitatively combined and compared with results of studies of mental health services without case management. Combined effect sizes and significance levels for 12 outcome domains were calculated. Analysis of homogeneity was used to explore differences between models. RESULTS: Forty-four studies were analyzed; 35 compared assertive community treatment or clinical case management with usual treatment, and nine directly compared assertive community treatment with clinical case management. Both types of case management were more effective than usual treatment in three outcome domains: family burden, family satisfaction with services, and cost of care. The total number of admissions and the proportion of clients hospitalized were reduced in assertive community treatment programs and increased in clinical case management programs. In both programs the number of hospital days used was reduced, but assertive community treatment was significantly more effective. Although clients in clinical case management had more admissions than those in usual treatment, the admissions were shorter, which reduced the total number of hospital days. The two types of case management were equally effective in reducing symptoms, increasing clients' contacts with services, reducing dropout rates, improving social functioning, and increasing clients' satisfaction. CONCLUSIONS: Both types of case management led to small to moderate improvements in the effectiveness of mental health services. Assertive community treatment had some demonstrable advantages over clinical case management in reducing hospitalization. PMID- 11058190 TI - Characteristics of managed behavioral health care organizations in 1996. AB - OBJECTIVE: Data from a survey of managed behavioral health care organizations were analyzed to describe characteristics of these firms as well as service utilization and revenues. METHODS: Six managed behavioral health care organizations fully completed a survey by the American Managed Behavioral Healthcare Association in which they reported 1996 data for their contracts. The contracts represented more than 16 million covered lives and accounted for approximately 13 percent of all individuals enrolled in managed behavioral health care organizations in 1996. RESULTS: More than three-quarters of the contracts (77.5 percent) were nonrisk. Plans described as network-based risk contracts, which represented 28.7 percent of covered members, accounted for 71.1 percent of revenues. The vast majority of reported contracts were with private employers or health maintenance organizations (HMOs); these contracts accounted for 76.8 percent of reported revenues. HMOs tended to place somewhat greater restrictions on outpatient psychotherapy and outpatient medication management visits than did other types of payers; the most common limit for HMO-related contracts was 20 outpatient visits a year, compared with 50 visits a year for other payer categories. HMO contracts also required higher copayments for outpatient visits. Utilization of services differed by payer type; for example, use of inpatient services ranged from.18 percent of covered members for contracts with private employers to.90 percent of covered members for Medicaid contracts. CONCLUSIONS: Overall rates of service utilization were lower than those reported in other recent studies of managed behavioral health care. The survey findings provide a starting point to guide further investigation in this area. PMID- 11058191 TI - Enlisting indigenous community supporters in skills training programs for persons with severe mental illness. AB - OBJECTIVE: This study evaluated the generalization of skills training for severely and persistently mentally ill individuals who were paired with indigenous supporters. The supporters monitored the individuals' environments and prompted them to use their skills. METHODS: A total of 85 individuals with severe and persistent mental illness received six months of skills training. Forty-five of the participants received support from an individual of their choosing. The other 40 participants did not have supporters. At the end of the six-month skills training period, the supporters' participation was officially terminated, although they were encouraged to remain in their role for as long as both parties felt comfortable. The effects of the support were measured in terms of interpersonal functioning, acquisition and retention of the skills, psychopathology, global functioning, and satisfaction. Several process measures were also collected. RESULTS: The support procedures were evaluated favorably by both patients and supporters. The interpersonal functioning of the group with supporters was found to be significantly better than that of the nonsupported group at six- and 12-month follow-ups. No differences were found between the groups in symptoms-which were minimal during the entire training period-or skills learning and retention. CONCLUSIONS: The effects of support are likely applicable for a variety of individuals, supporters, and facilities. Indirect evidence suggested the importance of providing support for the supporters. PMID- 11058192 TI - Gold Achievement Award. Supportive residential services to reunite homeless mentally ill single parents with their children: The Emerson-Davis Family Development Center in Brooklyn, New York City. PMID- 11058193 TI - Gold Achievement Award. A comprehensive treatment program helps persons with severe mental illness integrate into the community: MHA Village, Long Beach, California. PMID- 11058194 TI - Significant achievement award. A comprehensive program for treating profoundly autistic children--Center for Autistic Children, Philadelphia. PMID- 11058195 TI - Compliance with antidepressant medication among prison inmates with depressive disorders. AB - This study assessed correlates of antidepressant medication compliance among 5,305 inmates of the Texas Department of Criminal Justice prison system who were diagnosed as having a depressive disorder. Use of tricyclic antidepressants, male gender, and higher age were all positively associated with medication compliance scores. This investigation provided no evidence that broader use of selective serotonin reuptake inhibitors would improve adherence to pharmacologic treatment in this population. The results also suggest that correctional administrators may wish to target younger inmates and women with interventions to improve medication compliance. PMID- 11058196 TI - Suicidal ideation and the choice of advance directives by elderly persons with affective disorders. AB - The charts of 191 geriatric psychiatric inpatients with major depressive disorder or bipolar disorder were examined to determine demographic and psychiatric factors associated with the advance directive to have cardiopulmonary resuscitation (CPR) or not to have it in a life-threatening situation. Overall, 48 percent of patients wanted no CPR, 12 percent wanted CPR only, and 41 percent wanted all therapy performed. Patients with suicidal ideation (26 percent of the sample) were significantly more likely to choose not to have CPR. Sixty percent of patients with suicidal ideation chose not to have CPR. Patients with bipolar disorder and patients under age 70 were more likely to choose CPR. PMID- 11058197 TI - Therapists' contacts with family members of persons with severe mental illness in a community treatment program. AB - Thirty-six therapists at an urban community mental health center responded to a survey about contacts with family members of 214 clients with serious mental illness. For 61 percent of the clients, the therapists reported at least one past year contact with a family member or someone acting as a family member. Contacts were typically by telephone and often took place during crises. The focus was on problem solving rather than on providing family therapy. Therapists perceived significant benefit from the contacts, which were achieved with little effort on their part. The results suggest that informal-and perhaps nonbillable-brief services to families are common. Such informal services fall short of recommended best-practice standards. PMID- 11058198 TI - The mentally ill poor: rethinking ethics. PMID- 11058199 TI - Family-friendly services: a modest proposal. PMID- 11058200 TI - Jerusalem syndrome or paranoid schizophrenia? PMID- 11058203 TI - Keeping up to date. PMID- 11058204 TI - Do we need an annual canadian respiratory conference? PMID- 11058205 TI - Summary of Canadian guidelines for the initial management of community-acquired pneumonia: an evidence-based update by the Canadian Infectious Disease Society and the Canadian Thoracic Society. AB - Community-acquired pneumonia (CAP) is a serious illness with a significant impact on individual patients and society as a whole. Over the past several years, there have been significant advances in our knowledge and understanding of the etiology of the disease, and an appreciation of problems such as mixed infections and increasing antimicrobial resistance. The development of additional fluoroquinolone agents with enhanced activity against Streptococcus pneumoniae has been important as well. It was decided that the time had come to update and modify the previous CAP guidelines, which were published in 1993. The current guidelines represent a joint effort by the Canadian Infectious Disease Society and the Canadian Thoracic Society, and they address the etiology, diagnosis and initial management of CAP. The diagnostic section is based on the site of care, and the treatment section is organized according to whether one is dealing with outpatients, inpatients or nursing home patients. PMID- 11058206 TI - Multidrug-resistant tuberculosis: long term follow-up of 40 non-HIV-infected patients. AB - BACKGROUND: There has been a steady increase in referrals of patients with multidrug-resistant tuberculosis (MDR-TB) who are human immunodeficiency virus (HIV) negative. Between 1986 and 1999, 40 patients were admitted to the authors' institution, eight of whom were admitted between January and June 1999. The management of such individuals is difficult. Although they are a clinically and epidemiologically important group of patients, few reports detail their management. OBJECTIVES: To review the demographics, clinical management and long term outcome of 40 non-HIV-infected individuals with MDR-TB referred to the only specialized TB inpatient service in Ontario. PATIENTS AND METHODS: Clinical hospital records were reviewed. RESULTS: Twenty-one men and 19 women (mean age 41+/-18 years) were admitted between January 1986 and June 1999 with MDR-TB and negative serology for HIV. Thirty-eight patients (95%) were born outside of Canada. Twenty-six patients (65%) had a history of previous TB. All were symptomatic with productive cough and positive sputum smears for acid-fast bacilli. There was a delay of 4.5 months between the initial diagnosis of TB and the recognition of the presence of MDR-TB. Four patients (10%) had TB resistant to isoniazid and rifampin only. Over 50% of patients had TB with additional resistance to streptomycin, and over 40% had additional resistance to ethambutol. Coexisting resistance was also found in significant numbers for pyrazinamide, ethionamide, para-aminosalicylic acid and cycloserine. Bacteriological conversion was achieved in 34 patients (85%). Six patients underwent surgical resection for localized lung disease. Twenty-four patients completed treatment and have remained free of disease for 33+/-25 months. All five patients (12%) who died had longstanding disease before their referral. CONCLUSIONS: MDR-TB is most frequently seen among immigrants with a previous history of TB, especially if they come from countries in which TB is highly prevalent. Outcome can be improved by a more timely recognition of MDR-TB. PMID- 11058207 TI - Influence of asthma education on asthma severity, quality of life and environmental control. AB - BACKGROUND: Several studies have examined the influence of asthma education, focusing mainly on the use of health services. OBJECTIVES: To assess the influence of an asthma education program (AEP) on airway responsiveness, asthma symptoms, patient quality of life (QOL) and environmental control. DESIGN: A prospective, randomized, controlled study with parallel groups. SETTING: Three tertiary care hospitals in Quebec. POPULATION: One hundred and eighty-eight patients with moderate to severe asthma. INTERVENTION: After optimization of asthma treatment with inhaled corticosteroids, patients were randomly assigned to receive either an education program based on self-management (group E) or usual care (control group C). RESULTS: One year after an AEP, there was a significant decrease in the number of days per month without daytime asthma symptoms in group E only (P=0.03). Asthma daily symptom scores decreased significantly in group E in comparison with group C (P=0. 006). QOL scores improved markedly in both groups after treatment optimization during the run-in period (P<0.01). After an AEP, the QOL score increased further in group E patients in comparison with group C patients (P=0.04). The concentration of methacholine that induces a 20% fall in forced expiratory volume in 1 s (PC20) improved significantly in both groups (group E 1.2+/-1.1 to 2.4+/-0. 2, group C 1.5+/-1.2 to 2.4+/-1.3, P<0.01). After one year, 26 of 37 patients from group E sensitized to house dust mites (HDM) adopted the specific measures recommended to reduce their exposure to HDM, while none of the 21 subjects from group C did (P<0.001). Among the patients sensitized to cats or dogs, 15% of patients from group E and 23% of patients in group C no longer had a pet at home at the final visit (P>0.5). CONCLUSIONS: One year after the educational intervention, it was observed that the program had added value over and above that of optimization of medication and regular clinical follow ups. The education program was highly effective in promoting HDM avoidance measures but minimally effective for removing domestic animals, suggesting that more efficient strategies need to be developed for the latter. PMID- 11058208 TI - Surgical management of lung gangrene. AB - OBJECTIVE: To review the outcomes of five cases of pulmonary resection for lung gangrene. DESIGN: A retrospective chart review. SETTING: A tertiary referral centre. POPULATION STUDIED: Five patients who underwent pulmonary resection for lung gangrene between April and December 1999. MAIN RESULTS: Pathological confirmation of lung gangrene was obtained in all cases. Three patients were ventilator dependent. All five patients had ongoing sepsis despite antibiotic therapy. Additional indications for resection included bronchopleural fistula (two patients), empyema (three patients) and hemoptysis (one patient). In two cases, there was evidence of bilateral, diffuse necrotizing pneumonia, while in three cases the process was localized to one side. Computed tomography revealed cavitation in four cases and the absence of blood supply to the affected lung in one case. Surgical resection included wedge resection (one patient), lobectomy (two patients), bilobectomy (one patient) and pneumonectomy (one patient). In all cases, the bronchial stump was reinforced with an intercostal flap. Postoperative empyema occurred in two cases, one treated by thoracoscopic decortication, the other by percutaneous drainage. There were no instances of stump leak and no deaths. One patient remains ventilator dependent. CONCLUSIONS: Resection for lung gangrene is possible even in the setting of diffuse parenchymal changes and ventilator dependency. A computed tomography scan of the chest is important to make the diagnosis of lung gangrene and to plan operative management. Reinforcement of the bronchial stump is critical. PMID- 11058209 TI - Rhinoviruses as pathogens of the lower respiratory tract. AB - Rhinoviruses (RVs) are the most common upper respiratory pathogens, inducing the majority of common colds worldwide. RV-related morbidity, although significant cumulatively, has been considered trivial for the individual patient. However, recent strong epidemiological associations of RVs with asthma exacerbations, including severe episodes requiring hospitalization, indicate that RV infections can result in serious disease. Current evidence supports the possibility that RVs infect the lower airways, inducing a local inflammatory response. Such evidence suggests that the role of RVs in other lower respiratory diseases, such as pneumonia, bronchitis, bronchiolitis and cystic fibrosis, should be re-examined with polymerase chain reaction-based methodologies, which are considerably more sensitive than traditional, cell culture-based techniques. The mechanisms through which RVs induce lower airway disease are studied to understand the relative contributions of the epithelial, neurogenic and immune components in the antiviral response, and to permit the design and implementation of specific treatments. PMID- 11058210 TI - Difficult asthma: consider all of the possibilities. AB - Asthma is a common respiratory disease that can often be managed successfully. However, there are patients that do not respond to the maximum doses of standard therapy and subsequently have a reduced quality of life. Many factors can contribute to a failure to respond to treatment, and a comprehensive approach is important when assessing and evaluating these patients. This report describes a patient referred for 'difficult to control asthma' who had multiple emergency department visits and hospitalizations. In addition to a history of wheezing, spirometry showed impaired flow and vital capacity was reduced. Further investigation showed a normal total lung capacity, and a computed tomography scan revealed main bronchus blockage by a tumour, which was confirmed by bronchoscopy. This led to a surgical resection of a mucoepidermoid carcinoma. This case highlights the need to consider all possibilities during the evaluation of patients with difficult asthma. PMID- 11058211 TI - Leaving the first amendment to the courts: a pro-evolution strategy for concerned scientists. PMID- 11058212 TI - Anatomy at EB 2001 PMID- 11058213 TI - International anatomical/morphological meetings PMID- 11058214 TI - Capitol hill update fall 2000 PMID- 11058215 TI - Reemerging stress: supraorbital torus morphology in the mid-sagittal plane? PMID- 11058216 TI - Stressed out: masticatory forces and primate circumorbital form. PMID- 11058217 TI - Axon guidance at the midline of the developing CNS. AB - Bilaterally symmetric animals must be capable of transmitting information between the left and right sides of their body to integrate sensory input and to coordinate motor control. Thus, many neurons in the central nervous system (CNS) of a wide variety of higher organisms project so-called commissural axons across the midline. Interestingly, these axons are never observed to re-cross the midline. On the other hand, some neurons project axons that remain on their own (ipsilateral) side of the CNS, without ever crossing the midline. Recent studies demonstrate that specialized cells which reside at the ventral midline of the developing vertebrate spinal cord and Drosophila ventral nerve cord play critical roles in regulating the guidance of both crossing and non-crossing axons. For example, these cells secrete positively-acting guidance cues that attract commissural axons over long distances to the midline of the CNS. Furthermore, short-range interactions between guidance cues present on the surfaces of midline cells, and their receptors expressed on the surfaces of pathfinding axons, allow commissural axons to cross the midline and prevent ipsilaterally projecting axons from entering the midline. Remarkably, as commissural axons cross over to the opposite side of the CNS, the molecular composition of their surfaces is dynamically altered so that they become responsive to repulsive midline guidance cues that they had previously ignored. Thus, this exquisitely controlled guidance system prevents commissural axons from crossing the midline more than once. Strikingly, many of the molecular mechanisms that control midline guidance appear to be evolutionarily conserved. PMID- 11058219 TI - Forthcoming topics PMID- 11058218 TI - The new architectonics: an invitation to structural biology. AB - The philosophy of art might offer an epistemological basis for talking about the complexity of biological molecules in a meaningful way. The analysis of artistic compositions requires the resolution of intrinsic tensions between disparate sensory categories-color, line and form-not unlike those encountered in looking at the surfaces of protein molecules, where charge, polarity, hydrophobicity, and shape compete for our attentions. Complex living systems exhibit behaviors such as contraction waves moving along muscle fibers, or shivers passing through the growth cones of migrating neurons, that are easy to describe with common words, but difficult to explain in terms of the language of chemistry. The problem follows from a lack of everyday experience with processes that move towards equilibrium by switching between crystalline order and chain-like disorder, a commonplace occurrence in the submicroscopic world of proteins. Since most of what is understood about protein function comes from studies of isolated macromolecules in solution, a serious gap exists between what we know and what we would like to know about organized biological systems. Closing this gap can be achieved by recognizing that protein molecules reside in gradients of Gibbs free energy, where local forces and movements can be large compared with Brownian motion. Architectonics, a term borrowed from the philosophical literature, symbolizes the eventual union of the structure of theories-how our minds construct the world-with the theory of structures-or how stability is maintained in the chaotic world of microsystems. PMID- 11058220 TI - Is neuropathic pain caused by the activation of nociceptive-specific neurons due to anatomic sprouting in the dorsal horn? PMID- 11058221 TI - Distribution of the EP3 prostaglandin E(2) receptor subtype in the rat brain: relationship to sites of interleukin-1-induced cellular responsiveness. AB - The activation of neurosecretory neurons that express corticotropin-releasing hormone (CRH) in response to increased circulating levels of interleukin-1beta (IL-1beta) depends on prostaglandin E(2) (PGE(2)) acting locally within the brain parenchyma. To identify potential central targets for PGE(2) relevant to pituitary-adrenal control, the distribution of mRNA encoding the PGE(2) receptor subtype EP3 (EP3R) was analyzed in rat brain. Hybridization histochemistry revealed prominent labeling of cells in discrete portions of the olfactory system, iso- and hippocampal cortices, and subcortical telencephalic structures in the septal region and amygdala. Labeling over the midline, intralaminar, and anterior thalamic groups was particularly prominent. EP3R expression was enriched in the median preoptic nucleus and adjoining aspects of the medial preoptic area (MPO) implicated in thermoregulatory/febrile responses and sleep induction. EP3R expressing cells were also prominent in brainstem cell groups involved in nociceptive information processing/modulation (periaqueductal gray, locus coeruleus (LC), parabrachial nucleus (PB), caudal raphe nuclei), arousal and wakefulness (LC, midbrain raphe and tuberomammillary nuclei); and in conveying interoceptive input, including systemic IL-1 signals, to the endocrine hypothalamus (nucleus of the solitary tract (NTS) and rostral ventrolateral medulla [VLM]). Combined hybridization histochemical detection of EP3R mRNA with immunolocalization of IL-1beta-induced Fos protein expression identified cytokine sensitive, EP3R-positive cells in the medial NTS, rostral VLM, and, to a lesser extent, aspects of the MPO. These findings are consistent with the view that increased circulating IL-1 may stimulate central neural mechanisms, including hypothalamic CRH neurons, through an EP3R-dependent mechanism involving PGE(2) mediated activation of cells in the caudal medulla and/or preoptic region. PMID- 11058222 TI - Relationship of EP(1-4) prostaglandin receptors with rat hypothalamic cell groups involved in lipopolysaccharide fever responses. AB - The action of prostaglandin E(2) (PGE(2)) in the preoptic area is thought to play an important role in producing fever. Pharmacologic evidence suggests that, among the four subtypes of E-series prostaglandin (EP) receptors, i.e., EP(1), EP(2), EP(3), and EP(4), the EP(1) receptor mediates fever responses. In contrast, evidence from mice with EP receptor gene deletions indicates that the EP(3) receptor is required for the initial (<1 hour) fever after intravenous (i.v.) lipopolysaccharide (LPS). To investigate which subtypes of EP receptors mediate systemic infection-induced fever, we assessed the coexpression of Fos-like immunoreactivity (Fos-IR) and EP(1-4) receptor mRNA in nuclei in the rat hypothalamus that have been shown to be involved in fever responses. Two hours after the administration of i.v. LPS (5 microg/kg), Fos-IR was observed in the ventromedial preoptic nucleus, the median preoptic nucleus, and the paraventricular hypothalamic nucleus. In these nuclei, EP(4) receptor mRNA was strongly expressed and the Fos-IR intensely colocalized with EP(4) receptor mRNA. Strong EP(3) receptor mRNA expression was only seen within the median preoptic nucleus but Fos-IR showed little coexpression with EP(3) receptor mRNA. EP(2) receptor mRNA was not seen in the PGE(2) sensitive parts of the preoptic area. Although approximately half of the Fos-immunoreactive neurons also expressed EP(1) receptor mRNA, EP(1) mRNA expression was weak and its distribution was so diffuse in the preoptic area that it did not represent a specific relationship. In the paraventricular nucleus, EP(4) mRNA was found in most Fos-immunoreactive neurons and levels of EP(4) receptor expression increased after i.v. LPS. Our findings indicate that neurons expressing EP(4) receptor are activated during LPS induced fever and suggest the involvement of EP(4) receptors in the production of fever. PMID- 11058223 TI - Topographic order of retinofugal axons in a marsupial: implications for map formation in visual nuclei. AB - We studied axon order in the primary visual pathway and in nine retinorecipient nuclei of a small marsupial, the fat-tailed dunnart (Sminthopsis crassicaudata) using animals at postnatal day (P) 40 and P80. Dorsal, ventral, nasal, and temporal axons enter the optic nerve true to their retinal origin being respectively dorsal, ventral, medial, and lateral; the arrangement is retained to the chiasm. Dorsal and ventral axons maintain their respective locations within the chiasm but at the base of the contralateral optic tract undergo a 180 degrees axial rotation, thus reversing the dorsoventral axis with respect to the retina. The alignment is conserved along the optic tract with dorsal and ventral axons mapping directly into appropriate quadrants of each retinorecipient nucleus. Nasal and temporal axons remain segregated as they decussate and lie respectively superficially and deep along the optic tract but with some intermingling. Within each retinorecipient nucleus, the nasotemporal axis is clearly demarcated, being represented in either a rostrocaudal (ventral and dorsal lateral geniculate nuclei; lateral posterior, dorsal terminal, and pretectal nuclei) or caudorostral (medial terminal and caudal pretectal nuclei, intergeniculate nucleus and superior colliculus) direction. The results imply that the dorsoventral axis in the retinorecipient nuclei could be due to preordering within the pathway, whereas the nasotemporal axis is determined by target-based cues. Moreover, cues for the orientation of the nasotemporal axis within retinorecipient nuclei must be localised within individual nuclei rather than as a single organiser, as previously envisaged (Chung and Cooke [1978] Proc. R. Soc. Lond. B. 210:335-373). PMID- 11058224 TI - Changes in tactile stimuli-induced behavior and c-Fos expression in the superficial dorsal horn and in parabrachial nuclei after sciatic nerve crush. AB - Neurons in the superficial laminae of the dorsal horn are dominated by input from peripheral nociceptors. Following peripheral nerve injury, low threshold mechanoreceptive Abeta-fibers sprout from their normal termination site in laminae III/IV into laminae I-II and this structural reorganization may contribute to neuropathic tactile pain hypersensitivity. We have now investigated whether a sciatic nerve crush injury alters the behavioral response in rats to tactile stimuli and whether this is associated with a change in the pattern of c Fos expression in the dorsal horn and the parabrachial area of the brainstem. Sciatic nerve crush resulted in a patchy but marked tactile allodynia manifesting first at 3 weeks and persisting for up to 52 weeks. C-Fos expression in the dorsal horn and parabrachial region was never observed on brushing the skin of the sciatic nerve territory in animals with intact nerves, but was found after sciatic nerve crush with peripheral regeneration. We conclude that after nerve injury, low threshold mechanoreceptor fibers may play a major role in producing pain-related behavior by activating normally nociceptive-specific regions of the central nervous system such as the superficial laminae of the dorsal horn and the parabrachial area. PMID- 11058225 TI - Cellular localization of lipocalin-type prostaglandin D synthase (beta-trace) in the central nervous system of the adult rat. AB - We applied high-resolution laser-scanning microscopy, electron microscopy, and non-radioactive in situ hybridization histochemistry to determine the cellular and intracellular localization of lipocalin-type prostaglandin D synthase, the major brain-derived protein component of cerebrospinal fluid, and its mRNA in leptomeninges, choroid plexus, and parenchyma of the adult rat brain. Both immunoreactivity and mRNA for prostaglandin D synthase were located in arachnoid barrier cells, arachnoid trabecular cells, and arachnoid pia mater cells. Furthermore, meningeal macrophages and perivascular microglial cells, identified by use of ED2 antibody, were immunopositive for prostaglandin D synthase. In the arachnoid trabecular cells, the immunoreactivity for prostaglandin D synthase was located in the nuclear envelope, Golgi apparatus, and secretory vesicles, indicating the active production and secretion of prostaglandin D synthase. In the meningeal macrophages, prostaglandin D synthase was not found around the nucleus but in lysosomes in the cytoplasm, pointing to an uptake of the protein from the cerebrospinal fluid. Furthermore, the existence of meningeal cyclooxygenase (COX) -1 and COX-2 was investigated by Western blot, Northern blot, and reverse transcriptase-polymerase chain reaction (RT-PCR), and the colocalization of COX-2 and prostaglandin D synthase was demonstrated in virtually all cells of the leptomeninges, choroid plexus epithelial cells, and perivascular microglial cells, suggesting that these cells synthesize prostaglandin D(2) actively. Alternatively, oligodendrocytes showed prostaglandin D synthase immunoreactivity without detectable COX-2. The localization of lipocalin-type prostaglandin D synthase in meningeal cells and its colocalization with COX-2 provide evidence for its function as a prostaglandin D(2)-producing enzyme. PMID- 11058226 TI - Mapping of architectonic subdivisions in the macaque monkey, with emphasis on parieto-occipital cortex. AB - The intraparietal sulcus (IPS) of the macaque monkey contains numerous areas associated with different aspects of cortical function, including motor control as well as visual, somatosensory, vestibular, and possibly auditory processing. This study focuses largely on the architectonic organization of areas within and near the IPS, but also examines remaining portions of the hemisphere with which the IPS is interconnected. We charted the location of up to 72 architectonically distinct areas plus numerous architectonic zones in individuals over a region covering most of the cortical hemisphere. Identified cortical subdivisions (areas plus zones) were represented on computationally generated flat maps in relation to gyral and sulcal geography, thereby facilitating the analysis of consistent as well as variable aspects of the sizes and relative positions of subdivisions across animals. Using myelin and Nissl stains, plus immunohistochemical staining with the SMI-32 antibody, 17 architectonic subdivisions were identified that are largely or entirely contained in the intraparietal and parieto-occipital sulci. This includes four newly identified zones: a heavily myelinated lateral occipitoparietal zone, termed LOP; a strongly SMI-32 immunoreactive zone termed 7t (near the tip of the IPS); plus medial and lateral subdivisions (VIPm and VIPl) of ventral intraparietal area (VIP), which was previously regarded as an anatomically homogeneous area. Within the superior temporal sulcus, we identified a densely myelinated zone termed the dorso-posterior subdivision of the medial superior temporal area (MSTdp) that bordered middle temporal area (MT). We charted the extent of numerous other architectonically defined subdivisions throughout the cortical hemisphere by using criteria largely based on previous studies, but in some instances involving revised or expanded identification criteria. PMID- 11058227 TI - Corticocortical connections of visual, sensorimotor, and multimodal processing areas in the parietal lobe of the macaque monkey. AB - We studied the corticocortical connections of architectonically defined areas of parietal and temporoparietal cortex, with emphasis on areas in the intraparietal sulcus (IPS) that are implicated in visual and somatosensory integration. Retrograde tracers were injected into selected areas of the IPS, superior temporal sulcus, and parietal lobule. The distribution of labeled cells was charted in relation to architectonically defined borders throughout the hemisphere and displayed on computer-generated three-dimensional reconstructions and on cortical flat maps. Injections centered in the ventral intraparietal area (VIP) revealed a complex pattern of inputs from numerous visual, somatosensory, motor, and polysensory areas, and from presumed vestibular- and auditory-related areas. Sensorimotor projections were predominantly from the upper body representations of at least six somatotopically organized areas. In contrast, injections centered in the neighboring ventral lateral intraparietal area (LIPv) revealed inputs mainly from extrastriate visual areas, consistent with previous studies. The pattern of inputs to LIPv largely overlapped those to zone MSTdp, a newly described subdivision of the medial superior temporal area. These results, in conjunction with those from injections into other parietal areas (7a, 7b, and anterior intraparietal area), support the fine-grained architectonic partitioning of cortical areas described in the preceding study. They also support and extend previous evidence for multiple distributed networks that are implicated in multimodal integration, especially with regard to area VIP. PMID- 11058228 TI - Phototransduction molecules in the pigeon deep brain. AB - Photoreceptive areas responsible for the regulation of photoperiodicity have been localized in the brain of some avian species. We found that immunoreactivities to rhodopsin and the alpha-subunit of rod-type transducin (Gt(1)alpha) were colocalized in cerebrospinal fluid (CSF) contacting neurons in the pigeon lateral septum, a possible site for photoreception. Furthermore, RT-PCR analyses showed specific gene expression of both cGMP-phosphodiesterase beta-subunit and cone type cGMP-gated cation channel alpha-subunit in this region. These results suggest that several components in rod/cone photoreceptors compositely contribute to the deep encephalic phototransduction cascade. PMID- 11058229 TI - Olfactory projections in a chondrostean fish, Acipenser baeri: an experimental study. AB - The presence of extrabulbar primary olfactory projections has been well established in teleosts. In order to investigate the phylogeny of these projections and to compare their targets with those of the secondary olfactory projections, the connections of the olfactory epithelium and olfactory bulb were studied by means of tract-tracing methods in a chondrostean, Acipenser baeri. Primary olfactory projections mainly extend to the glomerular layer of the olfactory bulb, but a significant number of extrabulbar efferent fibers course to various telencephalic and diencephalic regions. Both extrabulbar primary and olfactory bulb projections course in diffuse pathways. Extensive overlap was observed between the targets of these extrabulbar primary olfactory fibers and those of the secondary efferent projections of the olfactory bulb, though the latter were more numerous and reached additional targets. Tracer application to the olfactory bulb also revealed a number of bulbopetal neurons in the ipsilateral and contralateral telencephalic areas, as well as crossed interbulbar projections. The presence in a chondrostean of important extrabulbar primary projections and their extensive overlap with secondary olfactory projections suggest that such projections are a derived characteristic of bony fishes. PMID- 11058230 TI - Allatotropin-like neuropeptide in the cockroach abdominal nervous system: myotropic actions, sexually dimorphic distribution and colocalization with serotonin. AB - Allatotropin (AT) was isolated from the moth Manduca sexta as a peptide stimulating biosynthesis of juvenile hormone in the corpora allata, but has also been shown to be cardioactive in the same species. Here, we have investigated the presence and biological activity of AT-like peptide in the cockroaches Leucophaea maderae and Periplaneta americana with focus on abdominal ganglia and their target tissues. An antiserum to M. sexta AT was used for immunocytochemical mapping of neurons in the abdominal ganglia. A small number of interneurons and efferent neurons were found AT-like immunoreactive (AT-LI) in each of the abdominal ganglia. A prominent sexual dimorphism was detected in the terminal abdominal ganglion: in L. maderae the male ganglion there are approximately 18 AT LI neurons with cell bodies posteriorly and efferent axons in the genital nerves; in the female ganglion 4-5 AT-LI cell bodies (with efferent axons) were found in the same region. Correlated with the extra efferents in males, the male accessory glands are richly supplied by AT-LI fibers and in females a less prominent innervation was seen in oviduct muscle. A similar dimorphism was seen in abdominal ganglia of P. americana. A sexual dimorphism was also detected in the abdominal ganglia A4-A6 of L. maderae. In each of these ganglia, approximately 8 10 large AT-LI neuronal cell bodies were found along the midline; in females these neurons have significantly larger cell bodies than in males. In both sexes, and both cockroach species, two large dorsal midline neurons were detected in A-5 and 6, which seem to send axons to the hindgut: the rectal pads of the hindgut are supplied by arborizing AT-LI axons. In males and females of both species, efferent AT-LI axons from midline neurons in A3-A6 supply the lateral heart nerves and other neurohemal release sites with arborizations. The efferent midline neurons of females contain colocalized serotonin-immunoreactivity. We tested the in vitro actions of M. sexta AT on muscle contractions in the L. maderae hindgut and the abdominal heart of both species. The frequency of contractions in the hindgut increased dose dependently when applying AT at 5 x 10(-8) to 5 x 10(-6) M (maximal response at 5 x 10(-7) M). Also the frequency of contractions of the heart increased by application of AT (threshold response at 5 x 10(-9) M). This effect was more prominent in males of both species (maximal response was a 35-40% increase in males and 10-20% in females). In conclusion, an AT-like peptide is present in neurons and neurosecretory cells of cockroach abdominal ganglia and seems to play a role in control of contractions in the hindgut and heart and also to have some function in male accessory glands and oviduct. PMID- 11058231 TI - Differential distribution of nerve terminals immunoreactive for substance P and cholecystokinin in the sympathetic preganglionic cell column of the filefish Stephanolepis cirrhifer. AB - Immunoreactivity for substance P and cholecystokinin-8 was examined in the nerve fibers in the central autonomic nucleus, a cell column for sympathetic preganglionic neurons, in the filefish Stephanolepis cirrhifer. Substance P immunoreactive fibers were distributed throughout the entire rostrocaudal extent, but were more abundant in the caudal part of the column, where substance P immunoreactive varicosities sometimes made contacts with the sympathetic preganglionic neurons. Cholecystokinin-8-immunoreactive fibers were found almost entirely in the rostral part of the column, where a dense network of varicosities was in close apposition to a considerable number of the sympathetic preganglionic neurons. Double labeling immunohistochemistry showed that substance P fibers and cholecystokin-8 fibers were entirely different, and distinct from serotonin immunoreactive fibers. By using immunoelectron microscopy, synaptic specialization was sometimes observed between the dendrites of preganglionic neurons and varicosities immunoreactive for substance P and cholecystokinin-8. Substance P- and cholecystokinin-8 fibers were seen from the descending trigeminal tract, through the dorsolateral funiculus and the ventral portion of the dorsal horn, to the central autonomic nucleus. After colchicine treatment, substance P-immunoreactive perikarya were found in the cranial and spinal sensory ganglia. These results suggest that the sympathetic preganglionic neurons of the filefish receive innervation by substance P fibers and cholecystokinin fibers, and that the former might be of primary sensory origin. Topographical distribution of cholecystokinin-8-immunoreactive terminals in the central autonomic nucleus along the rostrocaudal extent might underlie the differential regulation of sympathetic activity via a distinct population of sympathetic preganglionic neurons. PMID- 11058232 TI - Overview PMID- 11058233 TI - Contrast echocardiography: clinical utility. AB - This article reviews the advances made by the echocardiographic contrast agents from their first appearance in the early 1970s with homemade preparations up to the new generation of transpulmonary contrast agents made of small microbubbles capable of transversing the lung's capillary bed. The great progress in contrast agent development has kept pace with the progress made by echocardiographic equipment, thus making the study of myocardial perfusion in the clinical settings a near-future reality. This article also discusses the medical need that myocardial contrast echo has the potential to satisfy. PMID- 11058234 TI - Design of an ultrasound contrast agent for myocardial perfusion. AB - Myocardial contrast echography (MCE) has been a major research objective in cardiovascular ultrasound for almost two decades. The design of a contrast agent fulfilling the needs of MCE requires taking into consideration a number of points: a basic decision has to be made whether a deposit agent or a free-flowing agent would be more appropriate and whether an agent active at low/medium mechanical index (MI) is preferable to an agent active only at high MI; only a small percentage of the cardiac output enters the coronary microcirculation, which means that highly sensitive bubble detection methods, such as harmonic imaging or pulse inversion, are needed; the low velocity of blood in the microcirculation that leads to extensive bubble destruction during imaging means that intermittent imaging and/or an agent active at low MI is (are) required; the duration of the contrast effect must be sufficient to allow a complete examination and is affected by the rate of contrast administration; the performance of the contrast agent should not be equipment-dependent. The ultimate goal in MCE is to be able to quantify blood flow in the various segments to determine if adequate oxygenation is achieved. Ultrasound-mediated bubble destruction followed by the measurement of bubble replenishment kinetics opens new perspectives for quantification. SonoVue is a free-flowing ultrasound contrast agent made of sulphur hexafluoride microbubbles stabilized by a highly elastic phospholipid monolayer. SonoVue is able to produce myocardial opacification at a wide range of acoustic pressures and in particular at MIs as low as 0.1. Its performance is not equipment-dependent. Good results for myocardial opacification have been observed in all animal species tested (dogs, minipigs, rabbits), using continuous as well as intermittent imaging. Trials are in progress to demonstrate the clinical utility of SonoVue for rest and stress perfusion studies, in particular for the diagnosis of CAD, the detection of myocardial infarction, the assessment of the success of interventions and myocardial viability, and the detection of hibernating myocardium. PMID- 11058235 TI - Clinical experience with SonoVue in myocardial perfusion imaging. AB - Ultrasound-enhancing agents have the potential to evaluate myocardial perfusion, adding a new dimension to echocardiography. This article summarizes the clinical studies involving SonoVue, a new intravenous ultrasound contrast agent, in assessing myocardial perfusion. Safe and well tolerated, SonoVue coupled with echocardiography has the capability to identify perfusion abnormalities, as confirmed by scintigraphic imaging. While the optimal modalities for ultrasound perfusion assessment are not yet determined, numerous technical advances have been introduced: continuous infusion or slow intravenous administration of the agent, harmonic intermittent imaging, pulse inversion, background subtraction, color coding, and others. SonoVue is a promising new agent in the booming field of myocardial contrast echocardiography. PMID- 11058236 TI - Myocardial contrast echocardiography: clinical benefit and practical issues. AB - Real-time bedside evaluation of myocardial perfusion after intravenous application of microbubbles is the ultimate goal for contrast echocardiography. Over the past decade rapid evolution has occurred in the development of contrast agents, ultrasound equipment tailored to their detection, and image interpretation. This article offers a review of the basic concepts of the technique's background, contrast agent design, and imaging technology. The major clinical indications of myocardial contrast echocardiography are evaluation of acute ischemic syndromes, diagnosis of viable myocardium following AMI, and the detection of CAD using stress contrast perfusion imaging. Furthermore, the article addresses the most significant practical problems and suggested solutions to master those problems. As major new achievements are realistic expectations for the first decade of the twenty-first century, we conclude that the coupling of a new generation of contrast agents with innovative echocardiographic instrumentation will ultimately enable the full potential of myocardial contrast echocardiography to be realized which may revolutionize modern echocardiography. PMID- 11058395 TI - Deficiencies in US medical care. PMID- 11058396 TI - Deficiencies in US medical care. PMID- 11058398 TI - Volume contents and index PMID- 11058397 TI - Deficiencies in US medical care PMID- 11058399 TI - Editorial PMID- 11058400 TI - Conjugated linoleic acid: implications for human health. AB - Conjugated linoleic acid (CLA) is being sold as a panacea that has the capability of reducing or eliminating cancer, preventing heart disease, improving immune function, and altering body composition to treat obesity or build lean body mass. Unfortunately, there has been very little published human research on CLA. This review will examine the literature on CLA and discuss the animal research on which the above claims are made. The limited human studies will be presented with an evaluation of the potential uses of CLA for human health and disease. PMID- 11058401 TI - Action of ouabain on the contraction of ileal longitudinal muscle by manganese ions in a Ca(2+)-free, high-K(+), Na(+)-sufficient or Na(+)-deficient solution. AB - The action of ouabain, an external Na(+)-dependent inhibitor of active glucose transport, on the contraction by Mn(2+)in a Ca(2+)-free, Na(+)-sufficient or Na(+)-deficient medium was examined in ileal muscle. Ouabain ( 9 x 10(-5)m) inhibited the contraction by 5 mm Mn(2+)in a Ca(2+)-free, 60 mm K(+), Na(+) sufficient medium in ileal muscle. The addition of pyruvate or lactate, utilized independently in the external Na(+)in the pretreatment with ouabain, induced the dose-dependent contraction and manganese uptake by Mn(2+)in the Ca(2+)-free, 60 mm K(+), Na(+)-sufficient medium. These results suggest that Mn(2+)-induced ileal contraction in a high-K(+), Na(+)-sufficient medium is maintained by an active glucose transport, dependent on external Na(+). While, in contrast, Mn(2+) induced contraction in a high-K(+), Na(+)-deficient medium is maintained by an external Na(+)-independent glucose transport, and is insensitive to ouabain. PMID- 11058402 TI - Effect of vigabatrin and gabapentin on phenytoin pharmacokinetics in the dog. AB - This study was aimed at investigating whether or not the kinetics of intravenously administered phenytoin (PT) was altered by oral administration of vigabatrin (VGB) or gabapentin (GBP). A daily dose of PT (12 mgkg(-1)i.v.) was given to a group of five beagle dogs for a period of 1 week. On day eight, plasma samples were serially collected over 24 h, after administration of the PT dose. PT administration was continued, along with supplementary oral VGB (60 mgkg(-1)) for another week and then plasma samples were collected for analysis of PT levels. The same protocol was followed for the PT (12 mgkg(-1), i.v.)-GBP (300 mg caps., p.o.) study on a separate group (n= 5) of dogs. Orally administered GBP did not significantly alter the pharmacokinetic parameters of parenteral PT. However VGB markedly changed the drug's kinetics, as evidenced by a 31% (P= 0.015) reduction in total body clearance (CL) and an increase of over 45% in half life (t(1/2)), (P= 0.013) and area under the plasma PT concentration-time curve (AUC), (P= 0.044). GBP does not appear to have any pharmacokinetic interaction with PT, while coadministration of VGB and PT results in a marked reduction in systemic clearance of the latter in the dog. PMID- 11058403 TI - Bioequivalence of endogenous substances facing homeostatic equilibria: an example with potassium. AB - Oral administration of endogenous substances in most cases results in negligible net increases in baseline plasma concentrations, associated with high variability. This poses the problem of their bioequivalence. Using the data obtained from a bioequivalence investigation of potassium aspartate (test vs reference formulation), the authors demonstrate the inconsistency of bioequivalence based on plasma concentrations and standard methods. Potassium aspartate was given orally at a dose of 15.8 mmoles to 12 healthy volunteers as test and reference values according to a two-period, two-formulation, two sequence design. The individual net values of the area under the curve of plasma concentration (AUC) and cumulative urinary excretion (CUE), both obtained with the test formulation as post-dose minus baseline, were multiplied by 2, 3, 4, 5 and 6 and added to the baseline in order to simulate the administration of increasing single doses of the test, assuming dose-linear kinetics. Data generated with the test formulation were compared with original data of the reference according to 90% confidence intervals. With AUC, bioequivalence of test and reference formulations was demonstrated with 1 : 1, 2 : 1 and 3 : 1 test to reference dose ratios. With CUE only the 1 : 1 dose ratio comparison produced bioequivalence. The authors conclude that bioequivalence of endogenous substances conducted with standard procedures in most cases is a useless exercise. With potassium and more generally with drugs cleared via urine, urinary excretion would reflect the extent of absorption more faithfully than AUC. PMID- 11058404 TI - Foundations of pharmacokinetics. PMID- 11058405 TI - Area under the curve and bioavailability. AB - The Area Under the Curve (AUC) is proportional to the fraction absorbed only if the clearance is constant and the concentration uniform; in all other cases the bioavailability cannot be determined by comparing AUCs. PMID- 11058406 TI - Transmucosal transport of tobramycin incorporated in SLN after duodenal administration to rats. Part I--a pharmacokinetic study. AB - Tobramycin-loaded solid lipid nanospheres (SLN) were prepared and administered to rats into the duodenum; their behaviour was compared to that of tobramycin-loaded SLN administered intravenously (i.v.). A tobramycin control solution was also administered to rats. Tobramycin in solution is not absorbed by the gastrointestinal route, while tobramycin incorporated in the SLN is absorbed. A high concentration of tobramycin is still present in plasma 24 hours after the duodenal administration of tobramycin-loaded SLN. Tobramycin-loaded SLN administered i.v. showed a prolonged circulation time compared to the i.v. administered tobramycin solution. The AUC of tobramycin in SLN administered duodenally is higher than those of tobramycin in SLN and in solution administered i.v. PMID- 11058407 TI - Effects of salicylic acid in glutamate- and kainic acid-induced neurotoxicity in cerebellar granular cell culture of rats. AB - Glutamate (10(-7)m) and one of its non-NMDA receptor agonists, kainic acid (10( 4)m), were administered to rat cerebellar granular cell cultures, and the neuroprotective role of salicylic acid was examined. Glutamate induced 38.58 +/- 1.45% neuronal cell death while kainic acid induced only 21.4 +/- 2.01% despite being 1000 times more concentrated. The most effective dose for the neuroprotective effect of salicylate in glutamate-induced neurotoxicity was 10( 5)m and it had no protective effect at 10(-7)m. With kainic acid-induced toxicity, 10(-6)m salicylate had no protective effect but 10(-5)m and. 10(-4)m salicylic acid were very effective against kainic acid-induced toxicity. As an OH trapping agent, salicylate had a protective role in NMDA and non-NMDA receptor activated neuronal cell death. The present study gives some important clues about oxygen free radical generation having an important role in glutamate- and kainic acid-induced neurotoxicity. On the other hand, the neuroprotective effects of salicylic acid in the present study may depend on the pH alterations in salicylic acid solutions. PMID- 11058408 TI - A neurochemical study of the novel antiepileptic drug retigabine in mouse brain. AB - The novel antiepileptic drug, retigabine, has been reported to have multiple mechanisms of action, including potentiation of gamma -aminobutyric acid (GABA) and glutamate synthesis. We have investigated its effects on several GABA- and glutamate-related neurochemical parameters in mouse brain. Mice were administered retigabine either as a single dose or daily for 5 days. At 4 h after dosing, brains were removed and analysed for GABA, glutamate, and glutamine concentrations and for the activities of GABA-transaminase and glutamic acid decarboxylase. Single doses of retigabine significantly lowered brain concentrations of glutamate and glutamine. Repeated treatment significantly reduced the activity of GABA-transaminase. The drug was essentially without effect on all other parameters investigated. These results suggest that retigabine blocks GABA metabolism rather than enhancing GABA synthesis. In addition, the drug may also lower brain concentrations of the excitatory neurotransmitter glutamate and its precursor, glutamine. These effects may contribute to the antiepileptic action of retigabine. PMID- 11058409 TI - Inhibition of nicotinic receptor-mediated catecholamine secretion by Dryobalanops aromatica in bovine adrenal chromaffin cells. AB - Effect of the aqueous extract from a medicinal plant Dryobalanops aromatica(Dipterocarpaceae) on catecholamine secretion was investigated in bovine adrenal chromaffin cells. The aqueous extract inhibited [(3)H]norepinephrine ([(3)H]NE) secretion induced by 1,1-dimethyl-4-phenylpiperazinium iodide (DMPP), a nicotinic acetylcholine receptor (nAChR) agonist, with a half-maximal inhibitory concentration (IC(50)) of 8.4 +/- 1.7 microgml(-1). Increases in cytosolic calcium ([Ca(2+)](i)) and sodium ([Na(+)](i)) induced by DMPP were also inhibited by the extract. However, the binding of [(3)H]nicotine to nAChRs was not affected by the addition of the extract in receptor binding competition analysis, suggesting that active components in the extract and nicotine do not share the binding site in the nAChR. On the other hand, [Ca(2+)](i)increases induced by high K(+), ionomycin, bradykinin, angiotensin II, and thapsigargin were not inhibited by the extract. The data suggest that the extract from D. aromatica specifically inhibits catecholamine secretion by blocking nAChR in a noncompetitive manner. PMID- 11058410 TI - Protective effects of Mangifera indica L. extract, mangiferin and selected antioxidants against TPA-induced biomolecules oxidation and peritoneal macrophage activation in mice. AB - We compared the protective abilities of Mangifera indica L. stem bark extract (Vimang) 50-250 mgkg(-1), mangiferin 50 mgkg(-1), vitamin C 100 mgkg(-1), vitamin E 100 mgkg(-1)and beta -carotene 50 mgkg(-1)against the 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced oxidative damage in serum, liver, brain as well as in the hyper-production of reactive oxygen species (ROS) by peritoneal macrophages. The treatment of mice with Vimang, vitamin E and mangiferin reduced the TPA induced production of ROS by the peritoneal macrophages by 70, 17 and 44%, respectively. Similarly, the H(2)O(2)levels were reduced by 55-73, 37 and 40%, respectively, when compared to the control group. The TPA-induced sulfhydryl group loss in liver homogenates was attenuated by all the tested antioxidants. Vimang, mangiferin, vitamin C plus E and beta -carotene decreased TPA-induced DNA fragmentation by 46-52, 35, 42 and 17%, respectively, in hepatic tissues, and by 29-34, 22, 41 and 17%, in brain tissues. Similar results were observed in respect to lipid peroxidation in serum, in hepatic mitochondria and microsomes, and in brain homogenate supernatants. Vimang exhibited a dose-dependent inhibition of TPA-induced biomolecule oxidation and of H(2)O(2)production by peritoneal macrophages. Even if Vimang, as well as other antioxidants, provided significant protection against TPA-induced oxidative damage, the former lead to better protection when compared with the other antioxidants at the used doses. Furthermore, the results indicated that Vimang is bioavailable for some vital target organs, including liver and brain tissues, peritoneal exudate cells and serum. Therefore, we conclude that Vimang could be useful to prevent the production of ROS and the oxidative tissue damages in vivo. PMID- 11058411 TI - Stimulation of insulin secretion in clonal BRIN-BD11 cells by the imidazoline derivatives KU14r and RX801080. AB - The imidazoline derivatives KU14R and RX801080 have each been reported to antagonize imidazoline-stimulated insulin secretion. This study investigated the effects of a range of concentrations of both KU14R and RX801080 on insulin secretion from the clonal pancreatic beta cell line, BRIN-BD11. In the presence of a stimulatory (8.4 m m) glucose concentration, both KU14R (50-200 microm;P< 0.01 to P< 0.001) and RX801080 (50-200 microm;P< 0.01 to P< 0.001) were found to dose-dependently stimulate insulin secretion. The imidazoline efaroxan (200 microm) stimulated insulin secretion (P< 0.001) from BRIN-BD11 cells. This insulinotropic effect was significantly augmented by KU14R (100-200 microm;P< 0.01 to P< 0.001) and RX801080 (200 microm;P< 0.05). Insulin secretion from BRIN BD11 cells was also stimulated by the novel guanidine derivative BTS 67 582 (200 microm;P< 0.001). This secretagogue action was augmented both by KU14R (25-200 microm;P< 0.001) and by RX801080 (25-200 microm;P< 0.05 to P< 0.001). It is concluded that, rather than acting as antagonists of imidazoline-induced insulin secretion, the imidazoline derivatives KU14R and RX801080 are themselves potent insulinotropic agents. PMID- 11058412 TI - Subtype identification and functional properties of inositol 1,4, 5-trisphosphate receptors in heart and aorta. AB - One of the major mechanisms by which hormones elevate intracellular Ca(2+)levels is by generating the second messenger inositol 1,4, 5-trisphosphate (InsP(3)), which activates a Ca(2+)channel (InsP(3)receptor) located in the endoplasmic reticulum (ER). This study undertakes to identify the InsP(3)receptor subtypes (isoforms) in heart and aorta and to characterize their functional properties. The InsP(3)receptor isoforms were identified from rat heart and aorta tissues using both reverse-transcriptase polymerase chain reaction (RT-PCR) to assess the presence of mRNA for the different isoforms and immunochemistry using InsP(3)receptor isoform-specific antibodies. Functional studies included ligand binding experiments using [(3)H]InsP(3)and InsP(3)-induced Ca(2+)release studies using Fluo-3 as the Ca(2+)sensing dye. All three isoforms of the InsP(3)receptor were identified using RT-PCR and immunochemical analyses. [(3)H]InsP(3)binding studies using microsomes derived from these tissues showed that heart had a 3 fold lower abundance of InsP(3)receptors than aorta, while both have considerably lower abundance than the well characterized cerebellar microsomes. The affinity of the InsP(3)binding to the receptor was also different in the three tissues. In cerebellum the K(d)was 60 nM, while aorta had a much higher K(d)of 220 nM. Heart microsomes, appeared to show two classes of binding affinity with K(d)s of 150 nM and 60 nM. Furthermore, the effects of free [Ca(2+)] on [(3)H]InsP(3)binding levels were also different for the three tissues. InsP(3)binding to both cerebellar and aorta microsomes decreased by 90% and 60%, respectively, above 30 nM free [Ca(2+)], while InsP(3)binding to heart was relatively insensitive to changes in [Ca(2+)]. At maximal InsP(3)concentrations, aorta microsomes were able to release about 5% of the accumulated Ca(2+), compared to 25% by cerebellar microsomes. Heart microsomes, however, showed only very little InsP(3)-induced Ca(2+)release ( <0.5%). The EC(50)concentration for InsP(3)-induced Ca(2+)release was 1.2 micro M for aorta while that for cerebellum was 0.3 micro M. Known agonists of the cerebellar InsP(3)receptor such as 3-deoxy InsP(3)and adenophostin A were also able to mobilize Ca(2+)from aorta microsomes. In addition, the competitive antagonist heparin and the non-competitive antagonists of the cerebellar InsP(3)receptor, tetracaine and tetrahexylammonium chloride, were also able to block InsP(3)-induced Ca(2+)release from aorta microsomes. PMID- 11058413 TI - Alteration by cadmium of rat submandibular gland secretory function and the role of the l-arginine/nitric oxide pathway. AB - The effects of cadmium, l-arginine (nitric oxide precursor) and N(omega)-nitro-l arginine methyl ester (l -NAME) as a nitric oxide synthesis inhibitor and cotreatment of them on rat submandibular secretory function were studied. Pure submandibular saliva was collected intraorally by micro polyethylene cannula from anaesthetized rats using pilocarpine as secretagogue. Fourteen days treatment with 10 mg l(-1)cadmium as cadmium chloride in drinking water caused significant alterations on salivary function. Salivary flow rate, total protein concentration and amylase activity of saliva were decreased while secretion of calcium was increased by cadmium. Two weeks treatment of rats with l -arginine (2.25%) in drinking water caused an increase in submandibular gland weight. Flow rate was reduced by l-NAME. The total protein concentration of saliva was increased by l arginine while decreased by l-NAME. Calcium concentration of saliva was reduced by l-arginine and increased by l-NAME. Cotreatment of cadmium with l-arginine prevented cadmium-induced reduction of flow rate while l-NAME cotreatment potentiated cadmium-induced reduction of flow rate. l-arginine showed a preventive effect on cadmium-induced decrease of protein concentration and reached control levels. l-arginine potentiated cadmium-induced increase of saliva calcium concentration. It is confirmed that nitric oxide (NO) has a role in salivary gland function. It is also concluded that cadmium inhibitory effects on salivary gland function are modulated by the NO system as it is observed that the cadmium inhibitory effect on submandibular gland function is diminished by l arginine and extended by l-NAME. Considering the properties of cadmium substitution for calcium in many intracellular events, different types of alterations can be discussed. PMID- 11058414 TI - Thyroid hormone changes in males exposed to lead in the Buenos Aires area (Argentina). AB - The relationship between lead levels in blood (PbB) and hormones T3, T4, T4F and TSH were studied in 75 subjects exposed to lead at work. PbB levels in blood were determined by atomic absorption spectrophotometry and hormones by enzymoimmunoassay. Positive and significant correlations among thyroid hormones and TSH vs blood lead level in the ranges 8-50 and 26-50 micro g dl(-1)were found ('r' between 0.304 and 0.621 and 0.431 and 0.619, respectively). At PbB levels between 8 and 26 micro g dl(-1)significant correlation was only found for TSH (r= 0.731). In the range PbB 50-98 micro g dl(-1), 'r' was significant only for T3 ( 0.746) and T4 (-0.514). Significant differences were observed in T4 and T4F levels between exposed and non-exposed groups. The results obtained indicate the need for monitoring thyroid hormones and TSH levels in workers exposed to lead. PMID- 11058415 TI - A viral etiology for Ewing's sarcoma. AB - Despite the finding of characteristic somatic mutations in the tumor tissue and efforts to identify risk factors, the etiology of Ewing's sarcoma (ES) is still unknown. ES is very different from other childhood bone cancers. It rarely occurs in the black population and has no animal model. Recently studies indicate that ES may have a neural, not mesenchymal, origin. It has a distinctive unimodal age incidence peak at adolescence. Because its incidence curve pattern has a striking resemblance to that of DES-related clear cell adenocarcinoma of the vagina, an in utero exposure might be considered. Although in utero chemical and hormonal exposures have not been found to be associated with ES in epidemiologic studies, we suggest that its etiology could be an in utero viral infection. We hypothesize that the epidemiological characteristics of ES suggest an association with cytomegalovirus (CMV). PMID- 11058416 TI - Ursodeoxycholic acid to inhibit the growth of hepatic metastases. AB - Cancer can be resistant to apoptosis. If such cancers had apoptotic stimuli in their microenvironment, these stimuli might induce apoptosis in surrounding host cells. The majority of host cells at the peritumoral margins of liver metastases are undergoing apoptosis. Damage to the bile duct system may result in bile acid release, which may cause apoptosis in surrounding host cells. Metastatic cells may be releasing substances, such as transforming growth factor-beta 1 (TGF-beta 1), that cause apoptosis in surrounding host tissue. Ursodeoxycholic acid might inhibit the growth of hepatic metastases that are resistant to apoptotic stimuli such as bile acids and TGF-beta 1. Ursodeoxycholic acid decreases apoptosis caused by other bile acids and TGF-beta 1. Chemotherapy of hepatic metastases resistant to apoptosis might cause apoptosis more in peritumoral host cells than in cancer cells. Antiapoptotic therapy might be effective in cancer sensitive to apoptosis depending on its interactions with chemotherapy and tumor cells. PMID- 11058417 TI - Reduced cancer rates in agricultural workers: a benefit of environmental and occupational endotoxin exposure. AB - Numerous epidemiological studies have suggested reduced cancer rates in workers employed in agricultural industries. Traditionally, these observations have been attributed to low tobacco consumption and the healthy worker effect(s). Recent investigations have suggested that endotoxin may be responsible for reducing lung cancer rates in various occupational groups. Endotoxin anticancer properties are believed to be mediated through immunological mechanisms. This paper provides evidence and suggests a hypothesis for endotoxin-mediated reduced cancer rates in agricultural workers. PMID- 11058418 TI - Up-regulation of endothelial nitric oxide activity as a central strategy for prevention of ischemic stroke - just say NO to stroke! AB - Nitric oxide (NO) produced by the endothelium of cerebral arterioles is an important mediator of endothelium-dependent vasodilation (EDV), and also helps to prevent thrombosis and vascular remodeling. A number of risk factors for ischemic stroke are associated with impaired EDV, and this defect is usually at least partially attributable to a decrease in the production and/or stability of NO. These risk factors include hypertension, high-sodium diets, homocysteine, diabetes, visceral obesity, and aging. Conversely, many measures which may provide protection from ischemic stroke - such as ample dietary intakes of potassium, arginine, fish oil, and selenium - can have a favorable impact on EDV. Protection afforded by exercise training, estrogen replacement, statin drugs, green tea polyphenols, and cruciferous vegetables may reflect increased expression of the endothelial NO synthase. IGF-I activity stimulates endothelial NO production, and conceivably is a mediator of the protection associated with higher-protein diets in Japanese epidemiology and in hypertensive rats. These considerations prompt the conclusion that modulation of NO availability is a crucial determinant of risk for ischemic stroke. Multifactorial strategies for promoting effective cerebrovascular NO activity, complemented by measures that stabilize platelets and moderate blood viscosity, should minimize risk for ischemic stroke and help maintain vigorous cerebral perfusion into ripe old age. The possibility that such measures will also diminish risk for Alzheimer's disease, and slow the normal age-related decline in mental acuity, merits consideration. A limited amount of ecologic epidemiology suggests that both stroke and senile dementia may be extremely rare in cultures still consuming traditional unsalted whole-food diets. Other lines of evidence suggest that promotion of endothelial NO activity may decrease risk for age-related macular degeneration. PMID- 11058419 TI - Rapid-eye-movement sleep involves the memory-conversion circuits in a brain model. AB - People can remember the content of a dream in rapid eye movement (REM) sleep but cannot do so in slow-wave sleep. According to a brain model, memory is stored in encoding synapses as presynaptic axonal 'on-off' patterns and modulating synapses help encoding synapses convert short-term memory into long-term memory. These lead to the hypothesis that REM sleep involves modulating synapses of the memory conversion circuits including the anterior nuclei and dorsomedial nuclei of the thalamus. Cortical neurons get more rest in slow-wave sleep than in REM sleep. The locus coeruleus, raphe nuclei, and tuberomammillary nuclei get more rest during REM sleep when these nuclei cease to fire. The paralyses of peripheral muscles during REM sleep and cataplexy, and cessation of chorea, athetosis, hemiballismus, and parkinsonism tremor during sleep may result from spinal cord inhibition by the gigantocellular nuclei and raphe nuclei at the reticular formation. Sleep and wake relate to the light-dark cycle on the Earth. Were the light-dark cycle 50 hours a day, the human circadian clock might be around 50 hours. With increasing use of artificial light to keep people awake at night, it may affect the circadian rhythm and firing rate of neurons, the presynaptic axonal 'on-off' patterns as content of consciousness, and the mood. PMID- 11058420 TI - A nucleoside analogue of 2, 4-difluoropyridine has potential as an antiretroviral agent with multiple and unique mechanisms of action, and may be effective against the HIV organism. AB - The high rate of mutation which is inherent in reverse transcription of the HIV genome is a result of the lack of proof-reading function of the reverse transcriptase enzyme. This has allowed the HIV virus to develop resistance to multiple antiviral agents. It may be possible to use this viral property to advantage by treatment with an antiviral nucleoside analogue which is a close structural isostere of uridine and deoxyuridine. The drug is unable to form hydrogen bonds with adenine and will be excluded from host cell DNA by its 3' to 5' proof-reading exonuclease activity. However, reverse transcriptase, which has no such mechanism, will allow incorporation of the drug into proviral DNA. The drug will have an inhibitory effect on RNase H function. It will also be expected to cause delay in elongation at those sites in the template strand that contain two or more adjacent adenine bases, because two drug molecules will, for practical purposes, never be inserted in the same strand next to each other. The length of the delay in strand elongation will therefore be a function of the availability of the natural NTP or dNTP. Both the rate and fidelity of protein synthesis will be affected by the drug. There will be decreased stability of the proviral double stranded DNA and if the proviral DNA is able to integrate into the host cell chromosome, double stranded breaks may be produced by the host cells' DNA repair mechanisms. Finally there will be a specific 'strand trade' mutation that the drug will induce specifically into viral but not into cellular genetic material. PMID- 11058421 TI - Deficient detoxifying capacity in the pathophysiology of preeclampsia. AB - An imbalance between oxidative stress and maternal detoxification or antioxidant capacity may explain the symptoms of preeclampsia and the haemolysis-elevated liver enzymes-low platelets (HELLP) syndrome. Oxidative stress is known to induce damage of the endothelium, which is one of the pathophysiological features of preeclampsia and the HELLP syndrome. Administration of N-acetylcysteine, an antioxidant itself and a precursor of the endogenous antioxidant glutathione, might stabilize or even partly recover the process of endothelial damage and may lead to prolongation of pregnancy. PMID- 11058422 TI - Does longer-term memory storage never become overloaded, and would such overload cause Alzheimer's disease and other dementia? AB - According to an uncritically accepted axiom, the human brain's capacity for longer-term memory storage is never overloaded. This viewpoint is shown to be contrary to all evidence and beyond reasonable credibility. The entire currently obtainable evidence for or against overload comes via the presence or absence of its manifestation in behaviour. This manifestation would be as an incurable, deteriorating, specific form of memory disorder, associated with old age and consequent to increased data-inputting and reductions of brain capacity. There is in fact such a disorder, namely Alzheimer's disease and other dementia. This relationship provides elegant explanations for various peculiar findings, without encountering any counter-evidence. The premorbid accumulation of tangles in the hippocampus is explained via an integration of existing hippocampus theories. Long-term low doses of drugs that reduce memory formation could prevent or delay dementia. TV-watching, videos, and some memory-enhancing nootropic drugs could increase risk. PMID- 11058423 TI - Possible explanation of cephalic and noncephalic presentation during pregnancy: a theoretical approach. AB - This paper is based on fact that the fetus is exposed to gravity. The hypothesis is that from the 24th week of gestation an increasing percentage of fetuses occupies an exclusively cephalic presentation, since it allows an uncompromised posture in the caudal direction of body segments whose muscles are first affected by the occurrence and progressive increase of tone. Being in cephalic presentation, in a caudal direction, the fetus relieves body segments of the weight of the hypotonic-atonic part of the body in the cranial direction. In other words, cephalic presentation presents a body axis posture along the line of gravity. When the body axis posture along gravity is absent, the fetus simply fills the intrauterine cavity. In many such cases, the results are transverse lie, breech presentation and also cephalic presentation. PMID- 11058424 TI - Explaining the variety of human sexuality. AB - In this paper, the author formulates a theory to explain why human sexual orientation seems to run amok. The 'psychic instrument', as he terms it, is the baby's dreaming mind which interprets or misinterprets input from its sociocultural sexual environment. The baby, already born an omnisexual being, then develops a fantasy life with socially sanctioned or unsanctioned fetishes which are likely to be expressed when certain triggering situations arise. PMID- 11058425 TI - Body temperature as a risk factor for Alzheimer's disease. AB - A number of risk factors underlie the development of Alzheimer's disease. We propose low body temperature is also implicated. This is based on the belief that low temperature influences the biomechanics of the disease and promotes its development. Support for this hypothesis is found in a consideration of temperature effects on the disease process, in anecdotal observations and from our studies of people with Down syndrome. PMID- 11058426 TI - The relationship between thyroxine, oestradiol, and postnatal alopecia, with relevance to women's health in general. AB - Post-partum hair loss is possibly due to a reduction in the levels of oestradiol and thyroxine postnatally. Alopecia and/or a persistent loss of hair condition postnatally is associated with a group of symptoms (a syndrome), wherein postnatal depression is significant, as a result of physiologically inadequate levels of thyroxine (T4) and oestradiol (E2), secondary to physiological postnatal anterior pituitary dysfunction. Using this hypothesis, the author began to apply the same hypothesis to other female patients, who were not postpartum, but with similar symptomatology. The author became aware of the necessity for an adequate level of T4 to be present for correct oestrogenization to occur. He then goes on to hypothesize on the synergistic relationship that T4 and oestradiol may have in premenstrual syndrome (PMS), infertility, dysfunctional uterine bleeding, poor placental function, osteoporosis, and anorexia nervosa. He also discusses the role lowering T4 could play in the treatment of terminal cancer breast in premenopausal women. PMID- 11058427 TI - Resonant absorption of ultrasound energy as a method of HIV destruction. AB - A new method for eradicating human immunodeficiency virus (HIV) and other enveloped viruses is proposed. The method is based on the highly symmetric structure (e.g. icosahedral) of many viruses, which leads to a well-defined resonant frequency of ultrasound in the GHz range and which may be specifically absorbed by these structures and may subsequently lead to their irreversible damage. PMID- 11058428 TI - High-dose methylprednisolone may do more harm for spinal cord injury. AB - Because of the National Acute Spinal Cord Injury Studies (NASCIS), high-dose methylprednisolone became the standard of care for the acute spinal cord injury. In the NASCIS, there was no mention regarding the possibility of acute corticosteroid myopathy that high-dose methylprednisolone may cause. The dosage of methylprednisolone recommended by the NASCIS 3 is the highest dose of steroids ever being used during a 2-day period for any clinical condition. We hypothesize that it may cause some damage to the muscle of spinal cord injury patients. Further, steroid myopathy recovers naturally and the neurological improvement shown in the NASCIS may be just a recording of this natural motor recovery from the steroid myopathy, instead of any protection that methylprednisolone offers to the spinal cord injury. To our knowledge, this is the first discussion considering the possibility that the methylprednisolone recommended by NASCIS may cause acute corticosteroid myopathy. PMID- 11058429 TI - Insulin secretion as a potential determinant of homocysteine levels. AB - The recently reported association between insulin resistance and elevated plasma homocyst(e)ine may reflect the fact that hyperinsulinemia suppresses hepatocyte expression of cystathionine beta-synthase, as demonstrated in rats. Thus, other measures which enhance diurnal insulin secretion - such as a high-glycemic-index diet - can be expected to increase homocysteine levels. PMID- 11058430 TI - D-penicillamine in the neonatal period: possible beneficial effects on the AIDS associated infant mortality rate. PMID- 11058431 TI - Chronic fatigue syndrome and channelopathies. PMID- 11058432 TI - Family-based tests of association in the presence of linkage. AB - Linkage analysis may not provide the necessary resolution for identification of the genes underlying phenotypic variation. This is especially true for gene mapping studies that focus on complex diseases that do not exhibit Mendelian inheritance patterns. One positional genomic strategy involves application of association methodology to areas of identified linkage. Detection of association in the presence of linkage localizes the gene(s) of interest to more-refined regions in the genome than is possible through linkage analysis alone. This strategy introduces a statistical complexity when family-based association tests are used: the marker genotypes among siblings are correlated in linked regions. Ignoring this correlation will compromise the size of the statistical hypothesis test, thus clouding the interpretation of test results. We present a method for computing the expectation of a wide range of association test statistics under the null hypothesis that there is linkage but no association. To standardize the test statistic, an empirical variance-covariance estimator that is robust to the sibling marker-genotype correlation is used. This method is widely applicable: any type of phenotypic measure or family configuration can be used. For example, we analyze a deletion in the A2M gene at the 5' splice site of "exon II" of the bait region in Alzheimer disease (AD) discordant sibships. Since the A2M gene lies in a chromosomal region (chromosome 12p) that consistently has been linked to AD, association tests should be conducted under the null hypothesis that there is linkage but no association. PMID- 11058433 TI - Complex segregation analysis provides compelling evidence for a major gene underlying obsessive-compulsive disorder and for heterogeneity by sex. AB - Evidence from twin and family studies supports a genetic etiology for obsessive compulsive disorder (OCD). The purpose of this study was to test whether a major gene is implicated in a proportion of families with OCD. Complex segregation analyses of 153 families (80 case and 73 control), ascertained in the Johns Hopkins OCD Family Study, provided strong evidence for a major gene. A Mendelian dominant model, with significant sex effects and with residual familial effects, best explained the observed data. Stratification of the sample by the sex of probands provided further evidence of heterogeneity with respect to familial aggregation. Segregation analyses of 86 families with a female proband and of the 67 families with a male proband suggested that a Mendelian-dominant model with familial residual effects was the most parsimonious model explaining the inheritance of OCD in both subgroups. PMID- 11058434 TI - The Parkes lecture: controlled ovarian stimulation in women. AB - Recent advances in knowledge of the endocrine and paracrine mechanisms that regulate human ovarian folliculogenesis have been parallelled by the introduction into clinical practice of new drugs that can be used safely and effectively to stimulate ovarian function in infertile women. Most notably, recombinant DNA technology has been applied to the production of molecularly pure forms of the gonadotrophins, FSH and LH, opening the way to the development of improved strategies for manipulating the ovarian paracrine system. The clinical objectives of controlled ovarian stimulation fall into two categories, depending on patient needs: (1) induction of multiple follicles from which mature oocytes can be harvested for use in assisted reproduction protocols such as in vitro fertilization and embryo transfer; or (2) induction of spontaneous ovulation of a single mature follicle so that conception might occur in vivo. This review summarizes the physiological principles upon which the use of gonadotrophins for clinical purposes is based, highlighting new opportunities for improved treatment as a result of the availability of recombinant FSH and LH. PMID- 11058435 TI - Role of ovarian failure in reproductive senescence in aged red deer (Cervus elaphus) hinds. AB - Physiological and endocrine factors associated with reproductive senescence were assessed in a group of 19 ageing red deer hinds. Reproductive success, defined as the percentage of hinds weaning a calf successfully, decreased gradually from 89% at 6-7 years of age to 50% at 17 years, and subsequently decreased markedly; only one hind reared a calf at 19-20 years of age. When the 12 surviving hinds were approaching 21 years of age, they were compared with ten mature 7-year-old females over the onset of the breeding season. All hinds were subsequently killed, the reproductive tracts were recovered and antral (>/= 2 mm in diameter) and preantral follicle populations were determined by dissection (n = 7 hinds per age group) or stereological analysis (n = 2 ovaries per age group), respectively. Cyclical ovarian activity (plasma progesterone) was evident in fewer aged hinds compared with mature hinds (3/12 versus 10/10, P < 0.001) and mean plasma LH concentrations were higher in aged animals than in mature animals (0.57 +/- 0.05 and 0.20 +/- 0.05 ng ml(-1), P < 0.001). Mean uterine (44.2 +/- 4.5 and 75.4 +/- 4.2 g; P < 0.001) and ovarian masses (0.88 +/- 0.11 and 1.52 +/- 0.12 g; P < 0.001) were lower in the aged hinds, which also had fewer antral follicles than did mature hinds (0.89 +/- 0.35 and 23.5 +/- 4.5 follicles per hind, respectively; P < 0.001). Only one primordial follicle was observed in one of the ovaries of the aged hinds, compared with 7000-21 000 in the ovaries of mature hinds. The high gonadotrophin concentrations, paucity of primordial and antral follicles and failure of ovulation indicate collectively that waning reproductive performance after 17 years of age is primarily due to ovarian failure. PMID- 11058436 TI - Constitutive expression of heat shock proteins hsp25 and hsp70 in the rat oviduct during neonatal development, the oestrous cycle and early pregnancy. AB - Certain heat shock proteins are regulated by steroid hormones and are associated with oestrogen receptor function in reproductive tissues, indicating that these proteins have a role during implantation, decidualization and placentation. In the present study, the expression of hsp25, hsp70 and oestrogen receptor alpha were examined by immunohistochemistry in oviducts from rats during neonatal development, the oestrous cycle and during early pregnancy. Oestrogen receptor alpha was the first protein observed in the neonatal oviduct, and its expression preceded that of hsp70 and hsp25. Although these heat shock proteins have been associated with the oestrogen receptor, this study showed that during early development of the oviduct, the receptor protein was not associated with the concomitant expression of hsp25 and hsp70. However, these heat shock proteins were expressed when oviductal cells became differentiated. In the adult oviduct, hsp70 was more abundant than hsp25, moreover, there were no significant modifications in expression of hsp25 during the oestrous cycle. In contrast, the expression of hsp70 was significantly higher in epithelial cells during dioestrus, when the maximum amount of oestrogen receptor alpha was also observed. Therefore, the present study shows that hsp70, but not hsp25, is an oviductal protein modulated by the oestrous cycle and that it is a protein marker for specific phases of the oestrous cycle. In addition, hsp70 was more responsive to the hormonal changes in the infundibulum and ampullar regions of the oviduct. During early pregnancy, hsp25 expression was downregulated (unlike in the endometrium), whereas hsp70 was relatively abundant in the oviduct. hsp70 was observed in all functional segments of the oviduct during pregnancy, indicating that in the oviduct, this protein is modulated by oestrogens and progesterone and possibly by other pregnancy-related hormones. PMID- 11058437 TI - Effects of methylene blue, indigo carmine solution and autologous erythrocyte suspension on formation of adhesions after injection into rats. AB - The aim of this study was to determine whether autologous erythrocyte suspension can be used as a dye for evaluation of tubal patency and whether it has any advantages over methylene blue or indigo carmine solutions. Reproductively healthy female nulliparous Wistar Albino rats (n = 30), aged 6 months, mass 165 195 g, were assigned randomly to three groups. Rats received a 1 ml i.p. injection of 5% (w/v) methylene blue solution (methylene blue group: n = 10), 5% (w/v) indigo carmine solution (indigo carmine group: n = 10) or 5% (v/v) fresh autologous erythrocyte suspension (autologous erythrocyte group: n = 10). At 4 weeks after injection, a small sterile opening was made in the peritoneal cavity of each rat. The cavity was rinsed once with TCM-199 to collect macrophages. The rinsed peritoneal contents were cultured overnight to evaluate macrophage activation. The peritoneal opening was expanded for evaluation of adhesion formation. Only one rat from the autologous erythrocyte group had intra peritoneal adhesions (score 2), whereas all rats in the methylene blue group (score 1: n = 1; score 2: n = 4; score 3: n = 4; and score 4: n = 1) and seven rats in the indigo carmine group (score 1: n = 1; score 2: n = 2; score 3: n = 3; and score 4: n = 1) had intra-abdominal adhesions. Macrophage activity was observed in the cultured peritoneal contents collected from the methylene blue and indigo carmine groups but not from the autologous erythrocyte group. Adhesion formation could be due to macrophage activation caused by methylene blue and indigo carmine solutions. These results indicate that tubal patency can be observed by laparoscopy using autologous erythrocyte suspension. The results of this study are believed to be the first to indicate that a patient's own erythrocyte suspension could be used during observation of tubal patency by laparoscopy. However, further studies are required. PMID- 11058438 TI - Cloning of calves from various somatic cell types of male and female adult, newborn and fetal cows. AB - Twenty-four calves were cloned from six somatic cell types of female and male adult, newborn and fetal cows. The clones were derived from female cumulus (n = 3), oviduct (n = 2) and uterine (n = 2) cells, female and male skin cells (n = 10), and male ear (n = 5) and liver (n = 2) cells. On the basis of the number of cloned embryos transferred (n = 172) to surrogate cows, the overall rate of success was 14%, but based on the number of surrogate mothers that became pregnant (n = 50), the success rate was 48%. Cell nuclei from uterus, ear and liver cells, which have not been tested previously, developed into newborn calves after nuclear transfer into enucleated oocytes. To date, seven female and six male calves have survived: six of the females were from adult cells (cumulus (n = 3), oviduct (n = 2) and skin (n = 1) cells) and one was from newborn skin cells, whereas the male calves were derived from adult ear cells (n = 3), newborn liver and skin cells (n = 2), and fetal cells (n = 1). Clones derived from adult cells frequently aborted in the later stages of pregnancy and calves developing to term showed a higher number of abnormalities than did those derived from newborn or fetal cells. The telomeric DNA lengths in the ear cells of three male calves cloned from the ear cells of a bull aged 10 years were similar to those of the original bull. However, the telomeric DNA lengths from the white blood cells of the clones, although similar to those in an age-matched control, were shorter than those of the original bull, which indicates that telomeric shortening varies among tissues. PMID- 11058439 TI - Participation of reactive oxygen species in PGF2alpha-induced apoptosis in rat luteal cells. AB - Prostaglandin F(2alpha) (PGF(2alpha)) is implicated in the process of luteal regression in many species. Treatment of rat luteal tissue with PGF(2alpha) increases the generation of reactive oxygen species. Since reactive oxygen species have been implicated in apoptosis, the present study was undertaken to determine whether reactive oxygen species play a role in the PGF(2alpha)-induced apoptosis of rat luteal cells. Rat luteal cells were loaded with 6-carboxy-2, 7' dichlorodihydro-fluorescein (CDCFH) diacetate, di (acetomethyl ester), which can be oxidized by reactive oxygen species to yield CDCF, a fluorescent molecule, and the cells were treated with different doses of PGF(2alpha). Incubation with 100 micromol PGF(2alpha) l(-1) induced an increase in CDCF fluorescence (P < 0. 05). Treatment of cells with PGF(2alpha) for 48 h in serum-free medium induced a dose dependent increase in cell death, and these cells exhibited the morphological characteristics typical of apoptosis, including condensed or fragmented nuclei and fragmentation of internucleosomal DNA. Pretreatment of these cells with ascorbic acid, N,N'-dimethylthiourea, or superoxide dismutase, which acts as an antioxidant or a radical scavenger, prevented the PGF(2alpha)-induced apoptosis. These results demonstrate that PGF(2alpha) produces reactive oxygen species and induces apoptosis in rat luteal cells, indicating that the reactive oxygen species may induce apoptotic cell death during luteolysis. PMID- 11058440 TI - Protamine dissociation before decondensation of sperm nuclei during in vitro fertilization of pig oocytes. AB - The correlation between morphological changes and the dynamics of protamine in boar sperm chromatin during in vitro fertilization of pig oocytes matured in vitro was assessed. For this purpose, protamine was purified from boar sperm nuclei and an antiserum against protamine was developed. After affinity purification, the antiserum reacted exclusively with boar protamine during western blotting, showing no crossreactivity with core histones. Immunohistochemical evaluation revealed that only fully developed spermatid nuclei in boar testes stained strongly with the antiserum. When pig oocytes matured in vitro were fertilized in vitro, sperm penetration was observed in 37% of oocytes at 2 h after insemination and the penetration rate increased to 99% by 5 h after insemination, accompanied by an increase in polyspermic penetration. Paraffin wax sections of the inseminated oocytes were examined by immunohistochemical analysis with the antiserum. The proportion of condensed sperm nuclei that reacted with the antiserum was 87% of the sperm nuclei that penetrated by 2 h after insemination, and this decreased to 20 and 13% at 3 and 5 h after insemination, respectively. However, none of the decondensing sperm nuclei or male pronuclei reacted with the antiserum during the entire insemination period. These results indicate that a specific antiserum against boar protamine can be raised and, using this serum, it has been demonstrated that protamine is dissociated from boar sperm nuclei before decondensation during in vitro fertilization. PMID- 11058441 TI - Effect of long-term supplementation with arachidonic or docosahexaenoic acids on sperm production in the broiler chicken. AB - The possibility was investigated that dietary supplementation of the male chicken with long-chain polyunsaturated fatty acids of the n-6 and n-3 series may prevent the decrease in sperm output that normally occurs by 60 weeks of age. From 26 weeks of age, birds were raised on wheat-based diets supplemented with either maize oil (rich in linoleic acid, 18:2n-6), arasco oil (rich in arachidonic acid, 20:4n-6) or tuna orbital oil (rich in docosahexaenoic acid, 22:6n-3). The effects of the last two oils were investigated at two levels of vitamin E supplementation (40 and 200 mg kg(-1) feed). By 60 weeks of age, there was a small increase in the proportion of the main polyunsaturate of chicken sperm phospholipid, docosatetraenoic acid 22:4n-6, in chickens fed arasco oil diet compared with chickens given the maize oil diet, an effect that was potentiated at the higher dietary intake of vitamin E. Supplementation with tuna orbital oil significantly reduced the proportions of 20:4n-6 and 22:4n-6 in the sperm phospholipid and increased the proportion of 22:6n-3. The diet supplemented with tuna orbital oil and the lower level of vitamin E markedly depleted vitamin E from the tissues of the birds and decreased the concentration of vitamin E in the semen; these effects were largely prevented by the higher level of vitamin E in the diet. The susceptibility of semen to lipid peroxidation in vitro was increased in chickens fed arasco and tuna orbital oils with 40 mg vitamin E kg(-1) feed, but was reduced when 200 mg vitamin E kg(-1) feed was provided in the diet. The number of spermatozoa per ejaculate decreased by 50% between 26 weeks and 60 weeks of age in the birds fed the maize oil diet. This age-related decrease in the number of spermatozoa was almost completely prevented by feeding the birds with the oils enriched in either 20:4n-6 or 22:6n-3. Testis mass at 60 weeks of age was approximately 1.5 times greater in birds given of the arasco and tuna orbital oil diets compared with those given the maize oil diet. PMID- 11058442 TI - In vitro phagocytosis of boar spermatozoa by neutrophils from peripheral blood of sows. AB - A considerable number of spermatozoa are used in each sow in routine artificial insemination. However, within a few hours after insemination, many spermatozoa are phagocytosed by polymorphonuclear leucocytes. Some aspects of sperm transport in the female genital tract in the sow have been thoroughly investigated, whereas little is known about the mechanisms involved in the phagocytosis of spermatozoa, or about which spermatozoa (fresh, capacitated or dead) are the most susceptible to ingestion by polymorphonuclear leucocytes. In this study, phagocytosis was investigated by use of an in vitro phagocytosis assay. Polymorphonuclear leucocytes were challenged with either untreated, cold-shocked or frozen-thawed spermatozoa, or with spermatozoa that had been treated to induce capacitation in vitro. The influence of serum on phagocytosis was also investigated. Treatment of the semen to induce capacitation in vitro considerably reduced the phagocytosis of spermatozoa, whereas crude treatments like cold-shock or freezing and thawing reduced phagocytosis only in the first 15-30 min of incubation with polymorphonuclear leucocytes. Viable spermatozoa were phagocytosed mainly through a pathway that was independent of complement or other serum components (for example, antibodies). Complement had little effect on phagocytosis of spermatozoa, but did cause acrosomal exocytosis and cell death. PMID- 11058443 TI - Inhibition of pig granulosa cell adhesion and growth in vitro by immunoneutralization of epithelial cadherin. AB - Epithelial cadherin (E-cadherin) is a member of the cadherin family of calcium dependent cell adhesion molecules and is present in the ovary. Although expression of E-cadherin is high in healthy pig granulosa cells and low in granulosa cells of atretic follicles, the importance of E-cadherin-mediated adhesion in granulosa cell function is unclear. The aim of the present study was to determine the impact of immunoneutralization of E-cadherin on granulosa cell adhesion, DNA synthesis and cell proliferation in vitro. Before attachment, pig granulosa cells were exposed to a monoclonal E-cadherin antibody (DECMA-1) which blocks E-cadherin function. Controls included substitution of the antibody with either mouse ascites fluid or another E-cadherin antibody directed against the cytoplasmic domain and which was therefore inaccessible in intact cells. Both granulosa cell proliferation and insulin-like growth factor I-induced DNA synthesis were inhibited significantly in the presence of DECMA-1 compared with controls (P < 0.05). Control granulosa cells in culture formed large clusters with many cells packed tightly together. However, after 48 h exposure to the function-perturbing E-cadherin antibody, there was a significant decrease in the size of the granulosa cell clusters (P < 0.05) and the degree of cell-cell contact was reduced compared with control cultures. No effects on DNA synthesis, cell proliferation or cell adhesion were observed when DECMA-1 was substituted with either mouse ascites fluid or the antibody specific for the cytoplasmic domain of E-cadherin. In conclusion, these data provide evidence to support the hypothesis that E-cadherin is important for maintaining granulosa cell contact, DNA synthesis and cell proliferation in vitro. These results indicate that E cadherin plays a fundamental role in maintaining both the structure and function of ovarian follicles. PMID- 11058444 TI - Effect of granulocyte-macrophage colony-stimulating factor deficiency on ovarian follicular cell function. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF), a cytokine secreted by lymphohaemopoietic and other cell lineages, is known to influence ovarian cyclicity and embryo development. The aim of this study was to examine the effect of GM-CSF on ovarian follicular cell function using GM-CSF-deficient (GM -/-) mice. Immature GM -/- and GM +/+ mice were stimulated with eCG, and cumulus oocyte complexes and mural granulosa cells were collected 48 h later. Expression of GM-CSF receptor (GM-CSFR) alpha and beta mRNA subunits by cumulus-oocyte complexes and mural granulosa cells was examined using RT-PCR. Cumulus-oocyte complexes from both genotypes were found to express mRNA for the GM-CSFRalpha subunit only, while the mural granulosa cells expressed both the alpha and beta receptor subunits. Cumulus-oocyte complexes recovered from GM -/- mice had approximately twice the number of cumulus cells per cumulus-oocyte complex than did those of GM +/+ mice (P < 0.05), even though the growth-promoting activity of denuded GM -/- oocytes was found to be equivalent to that of wild-type oocytes. GM-CSF deficiency was associated with marginally increased DNA synthesis in cumulus cells and significantly (P < 0.05) lower progesterone production by mural granulosa cells recovered from GM -/- compared with those recovered from GM +/+ mice. The addition of rec-mGM-CSF in vitro did not affect DNA synthesis in either cell type or progesterone production by mural granulosa cells, irrespective of GM CSF status. There was no effect of GM-CSF deficiency on the capacity of FSH and insulin-like growth factor I to stimulate DNA synthesis in cumulus-oocyte complexes (approximately 15- and threefold, respectively) and in mural granulosa cells (approximately two- and threefold, respectively). Taken together, these data show that GM-CSF influences events associated with follicular maturation in mice. The effects of GM-CSF are not exerted directly in granulosa or cumulus cells, but appear to be mediated indirectly, perhaps through the agency of steroidogenesis-regulating secretions of local macrophage populations residing in the theca. PMID- 11058445 TI - Expression of mRNA encoding insulin-like growth factors I and II and the type 1 IGF receptor in the bovine corpus luteum at defined stages of the oestrous cycle. AB - Previous studies have implicated insulin-like growth factors I and II (IGF-I and II), in the regulation of ovarian function. The present study investigated the localization of mRNA encoding IGF-I and -II and the type 1 IGF receptor using in situ hybridization to determine further the roles of the IGFs within the bovine corpus luteum at precise stages of the oestrous cycle. Luteal expression of mRNA encoding IGF-I and -II and the type 1 IGF receptor was detected throughout the oestrous cycle. The expression of IGF-I mRNAvaried significantly during the oestrous cycle. IGF-I mRNA concentrations were significantly higher on day 15 than on day 10, and IGF-I mRNA in the regressing corpus luteum at 48 h after administration of exogenous prostaglandin was significantly greater than in the early or mid-luteal phase (days 5 and 10). In contrast, there was no significant effect of day of the oestrous cycle on expression of mRNA for IGF-II and the type 1 IGF receptor in the corpus luteum. Expression of IGF-II mRNA was localized to a subset of steroidogenic luteal cells and was also associated with cells of the luteal vasculature. mRNA encoding the type 1 IGF receptor was widely expressed in a pattern indicative of expression in large and small luteal cells. These data demonstrate that the bovine corpus luteum is a site of IGF production and reception throughout the luteal phase. Furthermore, this study highlights the potential of IGF-II in addition to IGF-I in the autocrine and paracrine regulation of luteal function. PMID- 11058446 TI - Expression of the lipocalin-type prostaglandin D synthase gene in the reproductive tracts of Holstein bulls. AB - The aim of this study was to localize expression of the prostaglandin D synthase gene in the reproductive tracts of Holstein bulls using northern blotting and in situ hybridization. For northern blotting, a digoxigenin-labelled prostaglandin D synthase cDNA probe was used to probe blots containing RNA isolated from the testes, epididymides, vas deferens, ampullae, seminal vesicles, prostate and bulbourethral glands of bulls. The digoxigenin-labelled cDNA for the bovine homologue of prostaglandin D synthase hybridized to a single band (approximately 0.9 kb) to RNA samples from the caput, corpus and cauda epididymides, as well as RNA samples from the vas deferens and the ampulla. The probe also detected a single band in testis samples, although the transcript size was slightly larger (approximately 1.0 kb) than the transcript found in the other tissues. The highest expression of prostaglandin D synthase was observed in the testes and caput epididymides. Prostaglandin D synthase transcripts were not found in the seminal vesicles or the prostate or bulbourethral glands using northern blotting. For in situ hybridization, antisense and sense riboprobes were synthesized and used to hybridize to cryosections obtained from the reproductive tissues of bulls. In situ hybridization of bull testes showed that prostaglandin D synthase transcripts were present within the germ cells in the adluminal compartment of the seminiferous tubules containing round and elongated spermatids, indicating that expression varied with stage of development of the seminiferous tubules. Prostaglandin D synthase expression was observed in the epithelial cells of the epididymides with greatest expression occurring in the caput epididymidis. Some expression was also observed in the epithelial cells of the vas deferens and a few cells of some lobules in the prostate and bulbourethral glands. Expression of the prostaglandin D synthase gene was not detected in ampullae or seminal vesicles by in situ hybridization. PMID- 11058447 TI - Magnetic resonance image attributes of the bovine ovarian follicle antrum during development and regression. AB - The magnetic resonance images and maps of bovine ovaries acquired at defined phases of follicular development and regression were studied to determine whether magnetic resonance image attributes of the follicular antrum reflect the physiological status of dominant and subordinate ovarian follicles. Ovariectomies were performed at day 3 of wave one, day 6 of wave one, day 1 of wave two and at >/= day 17 after ovulation. The timings of ovariectomies were selected to acquire growing, early static, late static and regressing follicles of the first wave and preovulatory follicles of the ovulatory wave. Pre-selection and subordinate follicles were also available for analysis. Serum samples were taken on the day of ovariectomy and follicular fluid samples were taken after imaging. Numerical pixel value and pixel heterogeneity in a spot representing approximately 95% of the follicular antrum were quantified in T(1)- and T(2)-weighted images. T(1) and T(2) relaxation rates (T(1) and T(2)), proton density, apparent diffusion coefficients and their heterogeneities were determined from the computed magnetic resonance maps. The antra of early atretic dominant follicles showed higher T(2) weighted mean pixel value (P < 0.008) and heterogeneity (P < 0. 01) and lower T(2) heterogeneity (P < 0.008) than growing follicles. Subordinate follicles in the presence of a preovulatory dominant follicle had higher T(1), T(1) heterogeneity, proton density, proton density heterogeneity, and lower mean pixel value in T(1)-weighted images than subordinate follicles of the anovulatory wave (P < 0.04). T(1) relaxation rate heterogeneity and proton density heterogeneity were positively correlated with follicular fluid oestradiol concentration (r = 0.4 and 0.3; P < 0.04). T(2) relaxation rate heterogeneity was positively correlated with follicular fluid progesterone concentration (r = 0.4; P < 0.008). Quantitative differences in magnetic resonance image attributes of the antrum observed among phases of follicular development and regression coincided with changes in the ability of the dominant follicle to produce steroid hormones and ovulate, and thus were indicative of physiological status and follicular health. PMID- 11058448 TI - Characterization of the glycoconjugates of boar testis and epididymis. AB - Lectin histochemistry was used to perform in situ characterization of the glycoconjugates present in boar testis and epididymis. Thirteen horseradish peroxidase- or digoxigenin-labelled lectins were used in samples obtained from healthy fertile boars. The acrosomes of the spermatids were stained intensely by lectins with affinity for galactose and N-acetyl-galactosamine residues, these being soybean, peanut and Ricinus communis agglutinins. Sertoli cells were stained selectively by Maackia ammurensis agglutinin. The lamina propria of seminiferous tubules showed the most intense staining with fucose-binding lectins. The Golgi area and the apical part of the principal cells of the epididymis were stained intensely with many lectins and their distribution was similar in the three zones of the epididymis. On the basis of lectin affinity, both testis and epididymis appear to have N- and O-linked glycoconjugates. Spermatozoa from different epididymal regions showed different expression of terminal galactose and N-acetyl-galactosamine. Sialic acid (specifically alpha2,3 neuraminic-5 acid) was probably incorporated into spermatozoa along the extratesticular ducts. These findings indicate that the development and maturation of boar spermatozoa are accompanied by changes in glycoconjugates. As some lectins stain cellular or extracellular compartments specifically, these lectins could be useful markers in histopathological evaluation of diseases of boar testis and epididymis. PMID- 11058449 TI - Ultrastructure of human blastocyst-endometrial interactions in vitro. AB - The interactions of seven human blastocysts with cultured endometrial cells were investigated by light microscopy and transmission electron microscopy. Trophoblastic-endometrial contact was observed at the lateral border of endometrial epithelial cells where trophoblast and endometrial epithelial cells shared apical junctional complexes and desmosomes. The first sign of penetration was invasion of a trophoblastic cytoplasmic protrusion between endometrial epithelial cells. In broad contact areas, lateral displacement of endometrial epithelial cells and formation of a peripheral pseudostratified epithelium were observed. When trophoblastic cells were interposed fully among endometrial epithelial cells, they formed a penetration cone and appeared to dislodge endometrial epithelial cells from the stromal compartment. A single penetration cone only was found in each specimen. Endometrial or trophoblastic degeneration was not observed. Formation of multinucleate (>/= three nuclei per cell) trophoblast cells was not observed, but many cells displayed areas with abrupt disappearance of well-defined plasma membranes, which is indicative of syncytium formation. In this study, adhesion and penetration occurred at the same time. The human blastocysts penetrated the endometrial surface epithelium by intrusive penetration. Epithelial penetration was achieved primarily by cellular syncytiotrophoblast-like cells and the first indications of syncytium formation were observed simultaneously with penetration of the epithelium. PMID- 11058450 TI - Immunohistochemical assessment of progesterone, oestrogen and glucocorticoid receptors in bovine placentomes during pregnancy, induced parturition, and after birth with or without retention of fetal membranes. AB - Steroid hormones play an important role in placental development. However, the exact cellular site of hormone action has not been evaluated in bovine placentomes. Thus, the present immunohistochemical study was designed to assess the distribution of progesterone receptors, oestrogen receptors and glucocorticoid receptors in bovine placentomes. Tissue specimens were obtained from cows at slaughter and from cattle during pre-term Caesarean section 27 h after prostaglandin administration, immediately after spontaneous parturition and from cattle that had retained the fetal membranes. Specific antibodies were used for receptor demonstration in tissue sections. Progesterone receptors were only detected in maternal connective tissue cells, whereas oestrogen receptors were also present in maternal crypt epithelium. At specific sites, both receptor immunoreactivities remained constant or changed significantly during pregnancy, were generally higher during Caesarean section and decreased post partum, but were less pronounced in cattle that released the fetal membranes than in those that retained the fetal membranes. Glucocorticoid receptors were evident in fetal connective tissue cells as well as in fetal and maternal blood vessels. Maternal crypt epithelial cells showed increasing immunoreactivities for glucocorticoid receptors during pregnancy. Receptor immunoreactivities tended to be lower after spontaneous parturition than during Caesarean section; these results were significant for progesterone and oestrogen receptors in animals that released the fetal membranes but not for those that retained the fetal membranes. The results indicate that in bovine placentome steroid hormone receptors are distributed in patterns that are specific to the type of cell, the stage of pregnancy and the tissue location, implying highly specific modulation of placental metabolism. Retention of the fetal membranes is reflected by altered placental receptor states at parturition. PMID- 11058451 TI - Ultrasonography and hormone profiles of persistent ovarian follicles (cysts) induced with low doses of progesterone in cattle. AB - The aims of this study were to expose dominant ovarian follicles at the end of the oestrous cycle to low progesterone concentrations similar to those that occur during stress, and to examine the effect of a subsequent small increase in progesterone 10 days later. Half a progesterone releasing intravaginal device (0.5 PRID) was administered to 13 heifers from day 15 of the oestrous cycle. In group 1 (n = 7), one 0.5 PRID remained in place until day 40 or until each heifer ovulated. In group 2 (n = 6), the first 0.5 PRID was removed on day 28, and replaced immediately with a second 0.5 PRID. Ultra-sonography and blood collection (10 ml) were conducted each day for 26 days from day 14 and then on alternate days. The largest follicle that emerged during the first 5 days after insertion of the initial 0.5 PRID remained > 10 mm in diameter for 15.3 +/- 1.7 and 11.6 +/- 0.4 days in groups 1 and 2, respectively. This period of dominance, during which no other follicles emerged, was closely correlated with the duration of plasma oestradiol concentrations exceeding 10 pg ml(-1). In four heifers from group 1, the persistent follicle ovulated between days 30 and 37 (sub-group 1a; 0.5 PRID expelled). In three heifers from sub-group 1b (0.5 PRID retained), the dominant follicle secreted oestradiol for 17 +/- 5 days but remained detectable by ultrasonography for a total of 33 +/- 8 days (range 26-52 days). Monitoring continued beyond day 40 in these animals. In group 2, the new 0.5 PRID inserted on day 28 resulted in an increase in plasma progesterone concentration of 0.9 +/- 0.3 ng ml(-1). Simultaneously, oestradiol decreased by 10.1 +/- 3.3 pg ml(-1), and a new follicular wave emerged 5-7 days later. In conclusion, exposure to very low concentrations of progesterone produced persistent follicles that secreted oestradiol for 17 days. This oestradiol production could be disrupted by a second increase of 0.9 ng ml(-1) in peripheral progesterone concentration. In the absence of the second progesterone treatment, some of the persistent follicles remained detectable by ultrasonography for up to 52 days, despite cessation of oestradiol secretion. PMID- 11058452 TI - Effect of progesterone on the activation of neurones of the supraoptic nucleus during parturition. AB - Parturition is driven by a pulsatile pattern of oxytocin secretion, resulting from burst firing activity of supraoptic oxytocin neurones and reflected by induction of Fos expression. Rats were injected with progesterone on day 20 of pregnancy to investigate the role of the decreasing progesterone:ratio oestrogen ratio, which precedes delivery, in the activation of supraoptic neurones. Progesterone delayed the onset of birth by 28 h compared with vehicle (control) and prolonged the duration of delivery, which was overcome by pulsatile injections of oxytocin, indicating that the slow delivery may reflect impaired oxytocin secretion. Parturient rats pretreated with progesterone had fewer Fos immunoreactive nuclei in the supraoptic nucleus than did parturient rats pretreated with vehicle. The number of Fos immunoreactive nuclei was not restored after oxytocin injection, indicating that appropriate activation of oxytocin neurones is impaired by progesterone and also that there is a lack of stimulatory afferent drive. Fos expression increased in the nucleus of the tractus solitarius during parturition in rats pretreated with either vehicle or progesterone, but not in rats that had been pretreated with progesterone and induced with oxytocin, indicating that this input was inhibited. Endogenous opioids inhibit oxytocin neurones in late pregnancy and the opioid antagonist, naloxone, increases Fos expression in supraoptic nuclei by preventing inhibition. However, progesterone attenuated naloxone-induced Fos expression in the supraoptic nucleus in late pregnancy and naloxone administered during parturition did not accelerate the duration of births delayed by progesterone administration, indicating that progesterone does not act by hyperactivation of endogenous opioid tone. RU486, a progesterone receptor antagonist, enhanced supraoptic neurone Fos expression in late pregnancy, indicating progesterone receptor-mediated actions. Thus, progesterone withdrawal is necessary for appropriate activation of supraoptic and tractus solitarius neurones during parturition. PMID- 11058453 TI - Regulation of spontaneous and induced resumption of meiosis in mouse oocytes by different intracellular pathways. AB - The mitogen-activated protein kinase-dependent and the cAMP-protein kinase A dependent signal transduction pathways were studied in cultured mouse oocytes during induced and spontaneous meiotic maturation. The role of the mitogen activated protein kinase pathway was assessed using PD98059, which specifically inhibits mitogen-activated protein kinase 1 and 2 (that is, MEK1 and MEK2), which activates mitogen-activated protein kinase. The cAMP-dependent protein kinase was studied by treating oocytes with the protein kinase A inhibitor rp-cAMP. Inhibition of the mitogen-activated protein kinase pathway by PD98059 (25 micromol l(-1)) selectively inhibited the stimulatory effect on meiotic maturation by FSH and meiosis-activating sterol (that is, 4,4-dimethyl-5alpha cholest-8,14, 24-triene-3beta-ol) in the presence of 4 mmol hypoxanthine l(-1), whereas spontaneous maturation in the absence of hypoxanthine was unaffected. This finding indicates that different signal transduction mechanisms are involved in induced and spontaneous maturation. The protein kinase A inhibitor rp-cAMP induced meiotic maturation in the presence of 4 mmol hypoxanthine l(-1), an effect that was additive to the maturation-promoting effect of FSH and meiosis activating sterol, indicating that induced maturation also uses the cAMP-protein kinase A-dependent signal transduction pathway. In conclusion, induced and spontaneous maturation of mouse oocytes appear to use different signal transduction pathways. PMID- 11058454 TI - Retention of cytoplasmic droplet by rat cauda epididymal spermatozoa after treatment with cytotoxic and xenobiotic agents. AB - Spermatozoa leaving the testis contain a cytoplasmic droplet which they release during transit through the epididymis before reaching the cauda epididymidis. The cytoplasmic droplet shows P450 aromatase activity, which plays a role in synthesis of oestrogen from androgen. In the present study, 3-month-old Wistar strain male albino rats were administered with the organophosphate insecticides malathion or dichlorvos, or the phytotherapeutics andrographolide or ursolic acid. Segments of the epididymis were subjected to histopathological and ultrastructural analyses and it was found that 60-95% of the spermatozoa residing in the lumen of the cauda epididymidis retained the cytoplasmic droplet. The motility of the spermatozoa released from the cauda epididymidis was inhibited. One of the mechanisms of action of these toxicants on male reproductive function may be attributed to the retention of the cytoplasmic droplet and the resultant impairment of sperm motility. PMID- 11058455 TI - Pulsatile GnRH secretion from primary cultures of sheep olfactory placode explants. AB - The aim of this study was to investigate the development of pulsatile GnRH secretion by GnRH neurones in primary cultures of olfactory placodes from ovine embryos. Culture medium was collected every 10 min for 8 h to detect pulsatile secretion. In the first experiment, pulsatile secretion was studied in two different sets of cultures after 17 and 24 days in vitro. In the second experiment, a set of cultures was tested after 10, 17 and 24 days in vitro to investigate the development of pulsatile GnRH secretion in each individual culture. This study demonstrated that (i) primary cultures of GnRH neurones from olfactory explants secreted GnRH in a pulsatile manner and that the frequency and mean interpulse duration were similar to those reported in castrated ewes, and (ii) pulsatile secretion was not present at the beginning of the culture but was observed between 17 and 24 days in vitro, indicating the maturation of individual neurones and the development of their synchronization. PMID- 11058456 TI - Identification of perivitelline N-linked glycans as mediators of sperm-egg interaction in chickens. AB - This study demonstrates that carbohydrates play an essential role in sperm-egg interactions in birds. Sperm-egg interaction was measured in vitro as the ability of spermatozoa to hydrolyse a small hole in the inner perivitelline layer, the equivalent of the mammalian zona pellucida. Preincubation with Triticum vulgaris lectin (WGA) and succinyl-WGA (S-WGA) at 10 microgram ml(-1) resulted in complete inhibition of sperm-egg interaction, whereas at the same concentration a range of other lectins (Canavalia ensiformis (Con A), Arachis hypogea (PNA), Ulex europaeus II (UEA II), Solanum tuberosum (STA), Tetragonolobus purpureas (LTA) and Pisum sativum (PSA)) were unable to inhibit sperm egg interaction significantly, although fluorescein-labelled derivatives of these lectins were found to stain the inner perivitelline layer. Significant inhibition of sperm-egg interaction was achieved by the addition of N-acetyl-D-glucosamine and fucoidin to the assay mixture; however, D-glucose, D-galactose, D-fucose and L-fucose had no significant effect on sperm-egg interaction. Pretreatment of the inner perivitelline layer with N-glycanase significantly reduced sperm-egg interaction, whereas treatment with O-glycanase had no effect. These results demonstrate that N-linked glycans play an essential role in sperm-egg interaction in chickens. PMID- 11058457 TI - Ultrasonography and hormone profiles of adrenocorticotrophic hormone (ACTH) induced persistent ovarian follicles (cysts) in cattle. AB - The objective of this study was to develop a model for the study of abnormal ovarian follicles in cattle by treating heifers with adrenocorticotrophic hormone (ACTH) (100 iu at 12 h intervals for 7 days, beginning on day 15 of the oestrous cycle). Cortisol concentrations increased (P < 0.05) within 24 h after beginning ACTH treatment and cortisol and progesterone concentrations remained elevated after cessation of ACTH treatment for 8 and 4 days, respectively. The pulses and surges of LH decreased during ACTH treatment, but FSH profiles were similar to those in controls and persistent or prolonged follicles were eventually observed in all heifers. In five heifers, prolonged dominant follicles ovulated after 10 days, whereas in six heifers, persistent follicular structures were present for 20 days, but ceased to secrete oestradiol after approximately 12 days. In the heifers with persistent follicular structures, new follicles emerged when the persistent follicle became non-oestrogenic. During the last 2 days of normal follicular growth, the concentration of oestradiol was greater than it was during prolonged or persistent follicle development (P < 0.05). There were no differences in the growth rates or maximum diameters of abnormal follicles that had different outcomes, but oestradiol concentrations were greater in prolonged follicles that ovulated compared with those follicles that persisted (P = 0.06). In conclusion, stimulation with ACTH resulted in a marked deviance from normal follicular activity. The aberrations were probably caused by the interruption of pulsatile secretion of LH (but not FSH) leading to decreased but prolonged oestradiol secretion. PMID- 11058458 TI - Follicular fluid rheology and the duration of the ovulatory process. AB - The fluid dynamics of ovulation were investigated to understand the mechanical role of follicular fluid in oocyte release. A set of equations describing the flow of fluid from an evacuating follicle was derived from basic principles. These equations demonstrate that, subject to assumptions about the available pressure differential and the source of the expulsive force, the size and shape of the ovulatory orifice have the largest influences on the rate of fluid loss, although the viscosity of the fluid is also an important variable. A thorough rheological examination of pig, bovine and human follicular fluids, performed using a cone-plate viscometer, demonstrated that these fluids have complex, non Newtonian characteristics. The fluids also undergo time-dependent and spontaneous changes in viscosity at constant shear rates; some fluids were subject to coagulation-like events. Viscosity characteristics were unrelated to broad parameters of follicle development. The models used representative viscosity values to demonstrate that variations in the rate and duration of follicle evacuation, as observed by ultrasonography, could be explained largely by variations in fluid viscosity and the characteristics of the ovulatory orifice. PMID- 11058459 TI - Uterine eosinophils and reproductive performance in interleukin 5-deficient mice. AB - Interleukin 5 is expressed in type 2 T lymphocytes and has a key role in driving the differentiation, recruitment and activation of eosinophils. Mice with a null mutation in the interleukin 5 gene (IL-5 -/- mice) have altered type 2 immune responses and severely depleted eosinophil populations. In the present study, the effect of interleukin 5 deficiency on the abundant population of eosinophils present in the female reproductive tract was investigated, and the reproductive performance in C57Bl/6 IL-5 -/- mice was measured. Endometrial eosinophils, detected on the basis of their endogenous peroxidase activity, were reduced in number by four-sevenfold during the oestrous cycle and in early pregnancy in IL-5 -/- mice. Eosinophils present in the cervix and decidual tissues at the time of parturition were similarly diminished. The temporal fluctuations in eosinophil recruitment and localization within these tissues were otherwise unchanged, indicating that interleukin 5 is not a necessary chemotactic agent in the female reproductive tract. Oestrous cycles were moderately greater in duration in IL-5 /- mice (mean +/- SD = 5.6 +/- 1.0 days in IL-5 -/- mice versus 5.0 +/- 0.8 days in IL-5 +/+ mice), owing to an extended period in oestrus (2.7 +/- 0.9 days per cycle in IL-5 -/- mice versus 1.8 +/- 0.7 in IL-5 +/+ mice). The interval between placing females with males and the finding of copulatory plugs was reduced significantly in interleukin 5-deficient mice. Implantation rates and subsequent fetal development were comparable in IL-5 -/- and IL-5 +/+ mice, irrespective of whether pregnancies were sired by syngeneic (C57Bl/6) or allogeneic (CBA or Balb/c) males, apart from a 10% increase in placental size and a 6.5% decrease in placental∶fetal ratio seen on day 17 in pregnancies sired by CBA males. Parturition and post-partum uterine repair were not compromised in interleukin 5 deficient mice, as judged by the length of gestation, and the outcomes of pregnancies initiated at post-partum oestrus. The birth weights and growth trajectories of pups were significantly influenced by interleukin 5 status, with small but significant increases in the weights of IL-5 -/- pups, particularly C57Bl/6 and CBA F(1) animals, remaining evident until adulthood. These data are consistent with the view that eosinophils have a role in endometrial tissue remodelling associated with the oestrous cycle, but indicate that the events of pregnancy and parturition proceed quite normally in the absence of maternal and fetal interleukin 5. However, strain-dependent effects of interleukin 5 deficiency on placental growth and function and subsequent weight gain in the newborn indicate that this cytokine may act through the maternal or fetal immune axis to exert subtle influences on reproductive outcome. PMID- 11058460 TI - Effect of TNF-alpha on LH and IGF-I modulated chicken granulosa cell proliferation and progesterone production during follicular development. AB - This study demonstrates the effects of recombinant human tumour necrosis factor a (rhTNF-alpha) and conditioned medium of the HD11-transformed chicken macrophage cell line on cultured chicken granulosa cells. Effects were studied on basal, IGF I- and LH-stimulated progesterone production and cell proliferation. Recombinant human TNF-alpha stimulated basal progesterone production in a dose-dependent manner in the granulosa cells of the largest follicle but had no effect on cells from the third largest follicle. TNF-alpha stimulated and sometimes inhibited progesterone production stimulated by IGF-I and LH alone or in combination depending on the size of the follicle and the concentration of LH or IGF-I applied. However, the inhibitory effect of TNF-alpha was significantly more pronounced in cells from the third largest follicle when high concentrations of IGF-I, LH or a combination of both were applied. TNF-alpha had no effect on basal cell proliferation in both the largest and the third largest follicles, but regulated responses to IGF-I and a combination IGF-I and LH in the cells of the third largest follicle but not those of the largest follicle. The data indicate that the normal hierarchy of follicles is maintained in the chicken ovary through the regulation of the activity of IGF-I and its interaction with LH. Conditioned medium of LPS-activated HD11 macrophages mimicked the effects of TNF-alpha and its interaction with IGF-I and LH on progesterone production and cell proliferation. The observation that the HD11-conditioned medium contained TNF alpha indicates that TNF-alpha produced by macrophages found in chicken follicles modulates granulosa cell growth and differentiation. PMID- 11058461 TI - Spermatogenesis and testicular tumours in ageing dogs. AB - Spermatogenesis was examined in testes from 74 dogs of various breeds without clinically detected testicular disease. A modified Johnsen score system was used to determine whether spermatogenesis deteriorates with ageing. The diameter of seminiferous tubules was measured in dogs without testicular disease to examine other possible effects of ageing on tubular performance. There appeared to be no relation between age and these variables. The influence of testicular tumours on spermatogenesis was also investigated in both affected and unaffected testes. The testes of 28 dogs with clinically palpable tumours and 21 dogs with clinically non-palpable tumours were investigated. In cases of unilateral occurrence of a tumour, impairment of spermatogenesis was observed only in the affected testis of dogs with clinically detected tumours. Bilateral occurrence of tumours, whether detected clinically or non-clinically, was associated with severe impairment of spermatogenesis. The prevalence of tumours increased during ageing. Eighty-six per cent of the clinically detected and 57% of the non-clinically detected tumours were found in old dogs. Multiple types of tumour and bilateral occurrence were very common. Seminomas and Leydig cell tumours were more frequent than Sertoli cell tumours. It was concluded that spermatogenesis per se did not decrease during ageing in dogs but the occurrence of testicular tumours increased with ageing and affected spermatogenesis significantly, as reflected by a lower Johnsen score. PMID- 11058462 TI - The needs for database research and for privacy collide. PMID- 11058463 TI - Protecting privacy while facilitating research. PMID- 11058464 TI - Delayed detection of psychosis: causes, consequences, and effect on public health. PMID- 11058465 TI - Large medical databases, population-based research, and patient confidentiality. AB - OBJECTIVE: This article is a discussion of the use of large clinical databases in population-based research on psychiatric disorders. METHOD: The authors review uses of large clinical databases in research on the etiology, impact, and treatment of psychiatric disorders. They also describe existing privacy safeguards applicable to use of medical records data in research. RESULTS: The growth of large medical databases has prompted increasing concern about the confidentiality of patient records. Efforts to restrict access to computerized medical data, however, may preclude use of such data in important and legitimate research. Prior research using large medical databases has made important contributions across a broad range of topics, including epidemiology, genetics, treatment effectiveness, and health policy. Continued population-based research will be essential in order to preserve the accessibility and quality of treatment for people with psychiatric disorders. CONCLUSIONS: Public domain research should be distinguished from proprietary or commercial uses of health information, and existing privacy safeguards should be vigorously applied. In our efforts to protect patient privacy, however, we should take care not to endorse or reinforce prejudices against psychiatric treatment and people who suffer from psychiatric disorders. Neither should we ignore important opportunities to improve quality of care and influence public policy through population-based research. PMID- 11058466 TI - Novel neurotransmitters and their neuropsychiatric relevance. AB - OBJECTIVE: The purpose of this review is to integrate insights regarding novel neurotransmitters or neuromodulators of neuropsychiatric significance. METHOD: Evolving concepts of neurotransmitter criteria are reviewed in light of the unexpected properties displayed by recently identified transmitters. RESULTS: Classic criteria for transmitters were based on the properties of acetylcholine but were markedly revised with the recognition of the catecholamines, serotonin, gamma-aminobutyric acid (GABA), and other amino acid transmitters and neuropeptides. Nitric oxide and carbon monoxide are notably atypical, as they are not stored in synaptic vesicles, are not released by exocytosis, and do not act at postsynaptic membrane receptor proteins. D-Serine, recently appreciated as the endogenous ligand for the glycine site of the glutamate N-methyl-D-aspartate (NMDA) receptor, overturns fundamental axioms of biology as well as those of neuroscience. It is a D-amino acid, and it is synthesized and stored in glia rather than neurons. Released glutamate acts on receptors on the protoplasmic astrocytes closely apposed to the synapse to release D-serine, which coactivates postsynaptic NMDA receptors together with glutamate. D-Serine is formed by serine racemase, which directly converts L-serine to D-serine. Inhibitors of this enzyme should reduce NMDA neurotransmission and might be therapeutic in stroke and other conditions associated with glutamate excitotoxicity. CONCLUSIONS: The diversity of novel neurotransmitters and venues of their activity afford multiple opportunities for therapeutic intervention. PMID- 11058467 TI - Images in neuroscience. Neural networks: neural systems IV: prefrontal cortex. PMID- 11058468 TI - Introspections. Charlie. PMID- 11058469 TI - "I am what I (don't) Eat": establishing an identity independent of an eating disorder. PMID- 11058471 TI - Presidential address: the doctor-patient relationship. PMID- 11058472 TI - Response to the presidential address: APA enters the twenty-first century. PMID- 11058474 TI - Neural correlates of imaginal aggressive behavior assessed by positron emission tomography in healthy subjects. AB - OBJECTIVE: Neurodegenerative or traumatic lesions of the frontal lobes often lead to abnormally aggressive behavior. The authors hypothesized that the imaginal evoking of scenarios involving aggressive behavior would be associated with a modulation of the functional activity in the human frontal cortex. METHOD: Regional cerebral blood flow (rCBF) determinations by positron emission tomography and psychophysiological measures of emotional responsivity were obtained in a group of 15 young healthy volunteers with good visual imagery abilities and no history of abnormal behavior while they imagined the same scenario with four variations involving emotionally neutral behavior and aggressive behavior. RESULTS: Compared to the imagined neutral scenario, the imagined scenarios involving aggressive behavior were associated with significant emotional reactivity and rCBF reductions in the ventromedial prefrontal cortex, suggesting that a functional deactivation of this cortical area occurs when individuals respond to the eliciting of imagined aggressive behavior. CONCLUSIONS: These results in healthy subjects further expand previous findings from animal and human studies by providing an in vivo functional demonstration of the involvement of the orbitofrontal cortex in the expression of aggressive behavior. They are also consistent with the hypothesis that a functional alteration of this cortical region may be present in individuals with pathological aggressive behavior. PMID- 11058475 TI - Feeling unreal: a PET study of depersonalization disorder. AB - OBJECTIVE: The goal of this study was to assess brain glucose metabolism and its relationship to dissociation measures and clinical symptoms in DSM-IV depersonalization disorder. METHOD: Positron emission tomography scans coregistered with magnetic resonance images of eight subjects with depersonalization disorder were compared to those of 24 healthy comparison subjects. The two groups did not differ in age, sex, education, performance on a baseline neuropsychological battery, or performance on a verbal learning task administered during [(18)F]fluorodeoxyglucose uptake. A cortical analysis by individual Brodmann's areas was performed. RESULTS: Compared to the healthy subjects, subjects with depersonalization disorder showed significantly lower metabolic activity in right Brodmann's areas 22 and 21 of the superior and middle temporal gyri and had significantly higher metabolism in parietal Brodmann's areas 7B and 39 and left occipital Brodmann's area 19. Dissociation and depersonalization scores among the subjects with depersonalization disorder were significantly positively correlated with metabolic activity in area 7B. CONCLUSIONS: Depersonalization appears to be associated with functional abnormalities along sequential hierarchical areas, secondary and cross-modal, of the sensory cortex (visual, auditory, and somatosensory), as well as areas responsible for an integrated body schema. These findings are in good agreement with the phenomenological conceptualization of depersonalization as a dissociation of perceptions as well as with the subjective symptoms of depersonalization disorder. PMID- 11058476 TI - Cue-induced cocaine craving: neuroanatomical specificity for drug users and drug stimuli. AB - OBJECTIVE: Cocaine-related cues have been hypothesized to perpetuate drug abuse by inducing a craving response that prompts drug-seeking behavior. However, the mechanisms, underlying neuroanatomy, and specificity of this neuroanatomy are not yet fully understood. METHOD: To address these issues, experienced cocaine users (N=17) and comparison subjects (N=14) underwent functional magnetic resonance imaging while viewing three separate films that portrayed 1 ) individuals smoking crack cocaine, 2) outdoor nature scenes, and 3) explicit sexual content. Candidate craving sites were identified as those that showed significant activation in the cocaine users when viewing the cocaine film. These sites were then required to show significantly greater activation when contrasted with comparison subjects viewing the cocaine film (population specificity) and cocaine users viewing the nature film (content specificity). RESULTS: Brain regions that satisfied these criteria were largely left lateralized and included the frontal lobe (medial and middle frontal gyri, bilateral inferior frontal gyrus), parietal lobe (bilateral inferior parietal lobule), insula, and limbic lobe (anterior and posterior cingulate gyrus). Of the 13 regions identified as putative craving sites, just three (anterior cingulate, right inferior parietal lobule, and the caudate/lateral dorsal nucleus) showed significantly greater activation during the cocaine film than during the sex film in the cocaine users, which suggests that cocaine cues activated similar neuroanatomical substrates as naturally evocative stimuli in the cocaine users. Finally, contrary to the effects of the cocaine film, cocaine users showed a smaller response than the comparison subjects to the sex film. CONCLUSIONS: These data suggest that cocaine craving is not associated with a dedicated and unique neuroanatomical circuitry; instead, unique to the cocaine user is the ability of learned, drug-related cues to produce brain activation comparable to that seen with nondrug evocative stimuli in healthy comparison subjects. PMID- 11058477 TI - Perfectionism in anorexia nervosa: variation by clinical subtype, obsessionality, and pathological eating behavior. AB - OBJECTIVE: The purpose of this study was to examine the role of perfectionism as a phenotypic trait in anorexia nervosa and its relevance across clinical subtypes of this illness. METHOD: The Multidimensional Perfectionism Scale and the perfectionism subscale of the Eating Disorder Inventory were administered to 322 women with a history of anorexia nervosa who were participating in an international, multicenter genetic study of anorexia nervosa. All participants were additionally interviewed with the Yale-Brown Obsessive Compulsive Scale and the Yale-Brown-Cornell Eating Disorder Scale. Mean differences on dependent measures among women with anorexia nervosa and comparison subjects were examined by using generalized estimating equations. RESULTS: Persons who had had anorexia nervosa had significantly higher total scores on the Multidimensional Perfectionism Scale than did the healthy comparison subjects. In addition, scores of the anorexia subjects on the Eating Disorder Inventory-2 perfectionism subscale exceeded Eating Disorder Inventory-2 normative data. For the anorexia nervosa participants, the total score on the Multidimensional Perfectionism Scale and the Eating Disorder Inventory-2 perfectionism subscale score were highly correlated. Total score on the Multidimensional Perfectionism Scale was also significantly related to the total score and the motivation-for-change subscale score of the Yale-Brown-Cornell Eating Disorder Scale. CONCLUSIONS: These data show that perfectionism is a robust, discriminating characteristic of anorexia nervosa. Perfectionism is likely to be one of a cluster of phenotypic trait variables associated with a genetic diathesis for anorexia nervosa. PMID- 11058478 TI - Involuntary treatment of eating disorders. AB - OBJECTIVE: Involuntary treatment of any psychiatric disorder has always been controversial, especially for eating disorders. Patients with an eating disorder of life-threatening severity frequently refuse hospitalization. In this study, the authors compared individual characteristics and treatment outcomes of patients admitted to an inpatient program for voluntary or involuntary treatment of their eating disorder. METHOD: This study examined 397 patients admitted to an inpatient treatment program over 7 years. Demographic measures, length of illness, weight history, and treatment response of patients admitted for voluntary treatment and those legally committed for involuntary treatment were compared. RESULTS: The two groups were similar in age, gender ratio, and marital status, but those legally committed for involuntary treatment had a longer illness duration and significantly more previous hospitalizations. At admission, the patients legally committed for involuntary treatment were lower in weight and required a significantly longer hospitalization to attain a healthy discharge weight. However, there was no statistically significant difference between involuntary and voluntary patients in rate of weight restoration (2.6 versus 2.2 lb/week, respectively). The groups did not differ in history of comorbid substance abuse or clinical depression but did differ significantly on all admission IQ measures. Eating disorder severity, as assessed by the Eating Attitudes Test-26, Eating Disorder Inventory, and MMPI-II, was similar for both patient groups. CONCLUSIONS: This study suggests that a substantial minority of patients with severe eating disorders will not seek treatment unless legally committed to an inpatient program. Despite the involuntary initiation of treatment, the short-term response of the legally committed patients was just as good as the response of the patients admitted for voluntary treatment. Further, the majority of those involuntarily treated later affirmed the necessity of their treatment and showed goodwill toward the treatment process. Only a long-term follow-up study will indicate whether these two populations differ in the enduring nature of their treatment response. PMID- 11058479 TI - Ionotropic glutamate receptor binding and subunit mRNA expression in thalamic nuclei in schizophrenia. AB - OBJECTIVE: Both thalamic and glutamatergic dysfunction have been implicated in the pathophysiology of schizophrenia. The authors examined ionotropic glutamate receptor expression in postmortem samples from patients with schizophrenia and comparison subjects, using the hypothesis that glutamate receptor expression differs in limbic nuclei of the thalamus in schizophrenia. METHOD: N-Methyl-D aspartate (NMDA), AMPA, and kainate receptor expression was determined in six thalamic nuclei from 12 subjects with DSM-III-R diagnoses of schizophrenia and eight psychiatrically normal individuals. The authors used in situ hybridization to determine NMDAR1, NMDAR2A-NMDAR2D, gluR1-gluR7, KA1, and KA2 subunit mRNA levels and receptor autoradiography to determine binding to glutamate binding sites of the three receptor subtypes and to the glycine, polyamine, and ion channel binding sites of the NMDA receptor. RESULTS: Glutamate receptor expression was lower at both transcriptional (NMDAR1, NMDAR2B, NMDAR2C, gluR1, gluR3, and KA2 subunit mRNAs) and posttranscriptional ([(3)H]ifenprodil and [(3)H]MDL105,519 binding to polyamine and glycine sites of the NMDA receptor) levels in the thalamus in patients with schizophrenia than in comparison subjects, but differences were most prominent in nuclei with reciprocal projections to limbic regions. CONCLUSIONS: Abnormalities in NMDA, AMPA, and kainate receptor expression in limbic thalamus are suggestive of the NMDA receptor hypoactivity hypothesis of schizophrenia and are consistent with diminished glutamatergic activity in the thalamus in schizophrenia. Alternatively, these results could suggest abnormal glutamatergic innervation in afferent and/or efferent regions, which are limbic structures that have been implicated in this illness. These results may provide a neurochemical anatomical substrate for antipsychotic therapies targeting ionotropic glutamate receptors. PMID- 11058480 TI - Lack of association between duration of untreated illness and severity of cognitive and structural brain deficits at the first episode of schizophrenia. AB - OBJECTIVE: The purpose of the study was to determine whether the duration of illness before antipsychotic drug treatment for schizophrenia was associated with the severity of cognitive deficits and volumetric brain structure anomalies observed in some patients with a first episode of schizophrenia. METHOD: Duration of psychotic symptoms and of other symptoms marking a behavioral change was estimated from structured interviews with 50 patients who had a first episode of schizophrenia and their family members. Interviews were conducted within a month of the patients' hospitalization. Duration of untreated psychotic symptoms and of behavioral change was correlated with neuropsychological summary scores from a comprehensive cognitive battery and with measurements of lateral ventricular, temporal lobe, and cerebral hemispheric volumes. RESULTS: No significant correlations were observed between measures of untreated illness and the severity of either cognitive or structural brain deficits at baseline. CONCLUSIONS: The duration of untreated symptoms of schizophrenia, for which an association with an uncontrolled toxic brain process has been proposed, is unlikely to explain why first-episode patients with schizophrenia have widespread deficits in cognitive functioning and have detectable ventricular enlargement and some loss of cortical mass. PMID- 11058481 TI - Features of structural brain abnormality detected in first-episode psychosis. AB - OBJECTIVE: Structural magnetic resonance imaging (MRI) studies that focus on first-episode psychosis avoid some common confounds, such as chronicity of illness, treatment effects, and long-term substance abuse. However, such studies may select subjects with poor short-term treatment response or outcome. In this study, the authors focus on structural brain abnormalities in never or minimally treated patients who underwent MRI scanning early in their first episode of psychosis. METHOD: The authors examined 37 patients (13 medication naive, 24 previously treated) who were experiencing their first episode of psychosis; the mean duration of symptoms was short (31 weeks). These patients were comparable in age, gender, handedness, ethnicity, and parental socioeconomic status to a group of 25 healthy comparison subjects. A three-dimensional, inversion recovery prepared, fast spoiled gradient/recall in the steady state scan of the whole brain that used 1.5-mm contiguous sections was performed to acquire a T(1) weighted data set. Human ratings of volumetric measurement of brain structures were performed with stereological techniques on three-dimensional reconstructed MRIs. RESULTS: The patient group had significant deficits in cortical gray matter, temporal lobe gray matter, and whole brain volume as well as significant enlargement of the lateral and third ventricles. Structural deviations were found in both treatment-naive and minimally treated subjects. No relationships were found between any brain matter volumes and positive or negative symptoms. CONCLUSIONS: Structural brain abnormalities were distributed throughout the cortex with particular decrement evident in gray matter. This feature is consistent with altered cell structure and disturbed neuronal connectivity, which accounts for the functional abnormality of psychosis. PMID- 11058482 TI - Nicotine transdermal patch and atypical antipsychotic medications for smoking cessation in schizophrenia. AB - OBJECTIVE: Schizophrenic patients have high rates of cigarette smoking. The authors compared the outcomes of two group psychotherapy programs for smoking cessation in patients with schizophrenia or schizoaffective disorder who were also treated with the nicotine transdermal patch and with either atypical or typical antipsychotic medications. METHOD: Forty-five subjects were randomly assigned to 1) the group therapy program of the American Lung Association (N=17) or 2) a specialized group therapy program for smokers with schizophrenia (N=28) that emphasized motivational enhancement, relapse prevention, social skills training, and psychoeducation. All subjects participated in 10 weeks of treatment with the nicotine transdermal patch (21 mg/day) and 10 weekly group therapy sessions and continued to receive their prestudy atypical (N=18) or typical (N=27) antipsychotic medications. Outcome variables included treatment retention, rate of smoking abstinence, and expired-breath carbon monoxide level. RESULTS: Smoking abstinence rates did not differ in the two group therapy programs. However, atypical antipsychotic agents, in combination with the nicotine transdermal patch, significantly enhanced the rate of smoking cessation (55.6% in the atypical agent group versus 22.2% in the typical group), which was reflected by a significant effect of atypical versus typical agents on carbon monoxide levels. Risperidone and olanzapine were associated with the highest quit rates. CONCLUSIONS: The results suggest that 1) smoking cessation rates with the nicotine transdermal patch are modest in schizophrenia, 2) specialized group therapy for schizophrenic patients is not significantly different from American Lung Association group therapy in its effect on smoking cessation, and 3) atypical agents may be superior to typical agents in combination with the nicotine transdermal patch for smoking cessation in schizophrenia. PMID- 11058483 TI - Sexual orientation in a U.S. national sample of twin and nontwin sibling pairs. AB - OBJECTIVE: Although previous studies have suggested that sexual orientation is influenced by familial factors, which may be partly genetic, these studies have relied on unrepresentative and potentially biased samples. The authors attempted to estimate the role of genetic and environmental factors in the determination of sexual orientation in a more representative sample. METHOD: Sexual orientation was assessed by a single item on a self-report questionnaire in a U.S. national sample of twin and nontwin sibling pairs. Sexual orientation was classified as heterosexual or nonheterosexual (bisexual or homosexual). The authors compared the similarity of sexual orientation in the monozygotic twins to the similarity in the same-sex dizygotic twins, all dizygotic twins, the same-sex dizygotic twins and sibling pairs, and all dizygotic twins and sibling pairs. Biometrical twin analyses were performed. RESULTS: All analyses demonstrated familial resemblance for sexual orientation. Resemblance was greater in the monozygotic twins than in the dizygotic twins or in the dizygotic twins plus nontwin siblings. Biometrical twin modeling suggested that sexual orientation was substantially influenced by genetic factors, but family environment may also play a role. No evidence was found for a violation of the equal-environment assumption regarding monozygotic and dizygotic twin pairs. CONCLUSIONS: Familial factors, which are at least partly genetic, influence sexual orientation. The results of these analyses should be interpreted in the context of low statistical power and the use of a single item to assess the complex phenotype of sexual orientation. PMID- 11058484 TI - Differentiation of homicidal child molesters, nonhomicidal child molesters, and nonoffenders by phallometry. AB - OBJECTIVE: The purpose of this study was to examine the ability of phallometry to discriminate among homicidal child molesters, nonhomicidal child molesters, and a comparison group of nonoffenders. METHOD: Twenty-seven child molesters who had committed or had attempted a sexually motivated homicide, 189 nonhomicidal child molesters, and 47 nonoffenders were compared on demographic variables and psychometrically determined responses to aural descriptions of sexual vignettes. Two phallometric indexes were used: the pedophile index and the pedophile assault index. The pedophile index was computed by dividing the subject's highest response to an aural description of sex with a "consenting" child by his highest response to description of sex with a consenting adult. The pedophile assault index was computed by dividing the subject's highest response to an aural description of assault involving a child victim by his highest response to description of sex with a "consenting" child. RESULTS: Homicidal child molesters, nonhomicidal child molesters, and nonoffenders were not significantly different in age or IQ. Homicidal and nonhomicidal child molesters had significantly higher pedophile index scores than nonoffenders. Significantly more homicidal child molesters (14 [52%] of 27) and nonhomicidal child molesters (82 [46%] of 180) than nonoffenders (13 [28%] of 47) had pedophile index scores equal to or greater than 1.0, but homicidal and nonhomicidal child molesters did not differ from each other. Significantly more homicidal child molesters (17 [63%] of 27) than either nonhomicidal child molesters (71 [40%] of 178) or nonoffenders (17 [36%] of 47) had pedophile assault index scores equal to or greater than 1.0, and nonhomicidal child molesters and nonoffenders were not significantly different from each other. Within-group analyses revealed that of the three groups, only the nonhomicidal child molesters exhibited a significant difference between their pedophile index scores and their pedophile assault index scores; their pedophile index scores were higher. CONCLUSIONS: Consistent with past research, the authors found that the pedophile index is useful in differentiating homicidal and nonhomicidal child molesters from nonoffenders and that the pedophile assault index is able to differentiate homicidal child molesters from nonhomicidal child molesters and nonoffenders. PMID- 11058485 TI - Mental disorders and the use of alternative medicine: results from a national survey. AB - OBJECTIVE: The study examined the relationship between mental disorders and the use of complementary and alternative medicine. METHOD: Data from a national household telephone survey conducted in 1997-1998 (N=9,585) were used to examine the relationships between use of complementary and alternative medicine during the past 12 months and several demographic variables and indicators of mental disorders. Structured diagnostic screening interviews were used to establish diagnoses of probable mental disorders. RESULTS: Use of complementary and alternative medicine during the past 12 months was reported by 16.5% of the respondents. Of those respondents, 21.3% met diagnostic criteria for one or more mental disorders, compared to 12.8% of respondents who did not report use of alternative medicine. Individuals with panic disorder and major depression were significantly more likely to use alternative medicine than those without those disorders. Respondents with mental disorders who reported use of alternative medicine were as likely to use conventional mental health services as respondents with mental disorders who did not use alternative medicine. CONCLUSIONS: We found relatively high rates of use of complementary and alternative medicine among respondents who met criteria for common mental disorders. Practitioners of alternative medicine should look for these disorders in their patients, and conventional medical providers should ask their depressed and anxious patients about the use of alternative medicine. More research is needed to determine if individuals with mental disorders use alternative medicine because conventional medical care does not meet their health care needs. PMID- 11058486 TI - Reliability and validity of DSM-IV axis V. AB - OBJECTIVE: The authors investigated the reliability and convergent and discriminant validity of the DSM-IV Global Assessment of Functioning Scale and two experimental DSM-IV axis V global rating scales, the Global Assessment of Relational Functioning Scale and the Social and Occupational Functioning Assessment Scale. METHOD: Forty-four patients admitted to a university-based outpatient community clinic were rated by trained clinicians on the three DSM-IV axis V scales. Patients also completed self-report measures of DSM-IV symptoms as well as measures of relational, social, and occupational functioning. RESULTS: The Global Assessment of Functioning Scale, Global Assessment of Relational Functioning Scale, and Social and Occupational Functioning Assessment Scale all exhibited very high levels of interrater reliability. Factor analysis revealed that the Global Assessment of Relational Functioning Scale and the Social and Occupational Functioning Assessment Scale are each more related to the Global Assessment of Functioning Scale individually than they are to each other. The Global Assessment of Functioning Scale was significantly related to concurrent patient responses on the SCL-90-R global severity index. The Social and Occupational Functioning Assessment Scale was significantly related to concurrent patient responses on the SCL-90-R global severity index and to a greater degree with both the Social Adjustment Scale global score and the Inventory of Interpersonal Problems total score. Although the Global Assessment of Relational Functioning Scale was not significantly related to any of the three self-report measures, it was related to the presence of clinician-rated axis II pathology. CONCLUSIONS: The three axis V scales can be scored reliably. The Global Assessment of Relational Functioning Scale and the Social and Occupational Functioning Assessment Scale evaluate different constructs. These findings support the validity of the Global Assessment of Functioning Scale as a scale of global psychopathology; the Social and Occupational Functioning Assessment Scale as a measure of problems in social, occupational, and interpersonal functioning; and the Global Assessment of Relational Functioning Scale as an index of personality pathology. The authors discuss further refinement and use of the three axis V measures in treatment research. PMID- 11058487 TI - Maintenance of brain monoamine oxidase B inhibition in smokers after overnight cigarette abstinence. AB - OBJECTIVE: The authors' goal was to replicate a previous finding that smokers have lower brain monoamine oxidase B (MAO-B) levels than comparison nonsmoking subjects and to determine if levels recover after overnight cigarette abstinence. METHOD: Brain MAO-B levels were measured by means of positron emission tomography in six smokers who were scanned twice: 11.3 hours (baseline) and 10 minutes after smoking one cigarette. RESULTS: Average MAO-B levels in smokers in the present study were similar to those found in the previous study and averaged 39% (SD=17) lower than those found in a comparison group of nonsmokers. Brain MAO-B levels did not differ between baseline levels and 10 minutes after smoking. CONCLUSIONS: This study reinforces the need to investigate whether MAO-B inhibition may account for some of the behavioral and epidemiological features of smoking. PMID- 11058488 TI - Treatment for depression following the 1996 National Depression Screening Day. AB - OBJECTIVE: Characteristics of the subsequent treatment received by people who screened positive for depression in the 1996 National Depression Screening Day were investigated. METHOD: A follow-up telephone survey was completed by 1,502 randomly selected participants from 2,800 sites. RESULTS: Of 927 people for whom additional evaluation was recommended, 602 (64.9%) obtained evaluations and 503 (83.6%) received treatment. Of these 503, 260 (51.7%) received psychotherapy and medication, 130 (25.8%) received medication only, and 93 (18.5%) received psychotherapy only. Compared with people without health or mental health insurance, individuals with health insurance (66.7% versus 57.5%) and mental health insurance (74.6% versus 55.3%) were more likely to comply with the recommendation to obtain follow-up evaluation. CONCLUSIONS: One-half of the people treated for depression received a combination of psychotherapy and medication. Lack of insurance was associated with not following the recommendation to obtain further evaluation and treatment. PMID- 11058489 TI - Disability pension for major depression in Finland. AB - OBJECTIVE: The authors' goal was to investigate the treatment received before receipt of a disability pension for major depression in a representative sample of depressed patients. METHOD: The medical statements for a random sample of 277 subjects drawn from the Disability Pension Register of the Social Insurance Institution were examined. The subjects selected represented individuals in Finland who were granted a disability pension because of DSM-III-R major depression during a 12-month period in 1993-1994. RESULTS: For 254 (92%) of the subjects, the statements regarding pension eligibility were written either by a psychiatrist or a psychiatric resident for patients who were currently being treated in psychiatric settings. There was an additional diagnosis of a comorbid mental disorder or a somatic disease contributing to disability in two-thirds of the statements. Overall, the statements indicated that 242 (87%) of the subjects were prescribed antidepressant medication, but only 24 (9%) received weekly psychotherapy, and only 11 (4%) received ECT. CONCLUSIONS: Most subjects granted a disability pension for major depression in Finland have comorbid mental or physical disorders contributing to their disability. Before receiving their pension, most received antidepressant treatment, but few received the established nonpharmacological treatments. PMID- 11058490 TI - Development and validation of a screening instrument for bipolar spectrum disorder: the Mood Disorder Questionnaire. AB - OBJECTIVE: Bipolar spectrum disorders, which include bipolar I, bipolar II, and bipolar disorder not otherwise specified, frequently go unrecognized, undiagnosed, and untreated. This report describes the validation of a new brief self-report screening instrument for bipolar spectrum disorders called the Mood Disorder Questionnaire. METHOD: A total of 198 patients attending five outpatient clinics that primarily treat patients with mood disorders completed the Mood Disorder Questionnaire. A research professional, blind to the Mood Disorder Questionnaire results, conducted a telephone research diagnostic interview by means of the bipolar module of the Structured Clinical Interview for DSM-IV. RESULTS: A Mood Disorder Questionnaire screening score of 7 or more items yielded good sensitivity (0.73) and very good specificity (0.90). CONCLUSIONS: The Mood Disorder Questionnaire is a useful screening instrument for bipolar spectrum disorder in a psychiatric outpatient population. PMID- 11058491 TI - Suicidal behavior in patients with current or past panic disorder: five years of prospective data from the Harvard/Brown Anxiety Research Program. AB - OBJECTIVE: The authors' goals were to examine predictors of suicidal behavior and provide guidelines for assessing suicide risk in patients with panic disorder. METHOD: Four hundred ninety-eight patients with panic disorder were followed for 5 years. Survival analysis was used to examine variables correlated with prospectively observed suicidal behavior. RESULTS: Subjects had a 0.06 probability of suicidal behavior during follow-up. Affective disorders, substance abuse, eating disorders, personality disorders, and being female were risk factors. Two subjects were suicidal in the absence of risk factors; both developed depression during follow-up. CONCLUSIONS: Panic disorder is not associated with suicidal behavior in the absence of other risk factors. PMID- 11058492 TI - Open trial of psychodynamic psychotherapy for panic disorder: a pilot study. AB - OBJECTIVE: This report contains preliminary data from an open trial of brief psychodynamic psychotherapy for panic disorder. METHOD: Fourteen patients with primary DSM-IV panic disorder completed a 24-session, twice-weekly course of psychodynamic psychotherapy. Other psychiatric treatment was not permitted throughout the 12-week treatment period and the 6-month follow-up. Symptoms were assessed at baseline, treatment termination, and 6-month posttermination follow up (40 weeks). RESULTS: Statistically significant, clinically meaningful improvements appeared in panic, depression, anxiety, and functional impairment both at treatment termination and at 6-month follow-up. CONCLUSIONS: Psychodynamic monotherapy can be used successfully to retain and treat patients with panic disorder. Psychodynamic interventions warrant further study for patients with panic disorder. PMID- 11058493 TI - Behavioral symptoms and psychiatric diagnoses among 162 children in nonalcoholic or alcoholic families. AB - OBJECTIVE: People with alcoholic relatives have high rates of alcohol abuse and dependence as adults, but their patterns of problems earlier in life are less clear. Many studies have not controlled for parental disorders other than alcoholism or for parents' socioeconomic status and general life functioning. The authors' goal was to conduct a study controlling for such factors. METHOD: Personal structured interviews and a behavioral checklist were administered to the parents of 162 children 7 years old or older whose fathers had participated in the 15-year follow-up of 453 sons of alcoholics with no history of antisocial personality disorder and sons of nonalcoholic comparison subjects originally selected from a university population. RESULTS: There was no significant relationship between a family history of alcoholism and childhood diagnoses of conduct, oppositional, or attention deficit disorders or with behavioral checklist summary scores. However, children with alcoholic relatives apparently have a slightly higher risk for drug abuse or dependence than those without alcoholic relatives. CONCLUSIONS: Once familial antisocial disorders and familial socioeconomic status are controlled for, a family history of alcoholism does not appear to relate to childhood externalizing disorders. PMID- 11058495 TI - Dr. Gardner replies PMID- 11058496 TI - New DSM-IV diagnosis of acute stress disorder. PMID- 11058494 TI - Amnesia after carbon monoxide poisoning. PMID- 11058497 TI - Testosterone replacement therapy for anxiety. PMID- 11058499 TI - The dopamine D(4) receptor gene and novelty seeking. PMID- 11058500 TI - Some ado about a polymorphism. PMID- 11058501 TI - Patients requesting psychiatric hospitalization. PMID- 11058503 TI - New DSM-IV diagnosis of acute stress disorder. PMID- 11058504 TI - New DSM-IV diagnosis of acute stress disorder. PMID- 11058505 TI - New DSM-IV diagnosis of acute stress disorder. PMID- 11058507 TI - New DSM-IV diagnosis of acute stress disorder. PMID- 11058523 TI - Leptin in pregnancy. AB - Leptin is a polypeptide hormone that aids in the regulation of body weight and energy homeostasis and is linked to a variety of reproductive processes in both animals and humans. Thus, leptin may help regulate ovarian development and steroidogenesis and serve as either a primary signal initiating puberty or as a permissive regulator of sexual maturation. Perhaps significantly, peripheral leptin concentrations, adjusted for adiposity, are dramatically higher in females than in males throughout life. During primate pregnancy, maternal levels that arise from adipose stores and perhaps the placenta increase with advancing gestational age. Proposed physiological roles for leptin in pregnancy include the regulation of conceptus growth and development, fetal/placental angiogenesis, embryonic hematopoiesis, and hormone biosynthesis within the maternal fetoplacental unit. The specific localization of both leptin and its receptor in the syncytiotrophoblast implies autocrine and/or paracrine relationships in this endocrinologically active tissue. Interactions of leptin with mechanisms regulating pre-eclampsia and maternal diabetes have also been suggested. Collectively, therefore, reports suggest that a better understanding of the regulation of leptin and its role(s) throughout gestation may eventually impact those causes of human perinatal morbidity and mortality that are exacerbated by intrauterine growth retardation, macrosomia, placental insufficiency, or prematurity. PMID- 11058524 TI - Prostaglandin metabolism in the ovine corpus luteum: catabolism of prostaglandin F(2alpha) (PGF(2alpha)) coincides with resistance of the corpus luteum to PGF(2alpha). AB - To examine possible mechanisms involved in resistance of the ovine corpus luteum to the luteolytic activity of prostaglandin (PG)F(2alpha), the enzymatic activity of 15-hydroxyprostaglandin dehydrogenase (PGDH) and the quantity of mRNA encoding PGDH and cyclooxygenase (COX-2) were determined in ovine corpora lutea on Days 4 and 13 of the estrous cycle and Day 13 of pregnancy. The corpus luteum is resistant to the action of PGF(2alpha) on Days 4 of the estrous cycle and 13 of pregnancy while on Day 13 of the estrous cycle the corpus luteum is sensitive to the actions PGF(2alpha). Enzymatic activity of PGDH, measured by rate of conversion of PGF(2alpha) to PGFM, was greater in corpora lutea on Day 4 of the estrous cycle (P < 0.05) and Day 13 of pregnancy (P < 0.05) than on Day 13 of the estrous cycle. Levels of mRNA encoding PGDH were also greater in corpora lutea on Day 4 of the estrous cycle (P < 0. 01) and Day 13 of pregnancy (P < 0.01) than on Day 13 of the estrous cycle. Thus, during the early estrous cycle and early pregnancy, the corpus luteum has a greater capacity to catabolize PGF, which may play a role in the resistance of the corpus luteum to the actions of this hormone. Levels of mRNA encoding COX-2 were undetectable in corpora lutea collected on Day 13 of the estrous cycle but were 11 +/- 4 and 44 +/- 28 amol/microgram poly(A)(+) RNA in corpora lutea collected on Day 4 of the estrous cycle and Day 13 of pregnancy, respectively. These data suggest that there is a greater capacity to synthesize PGF(2alpha), early in the estrous cycle and early in pregnancy than on Day 13 of the estrous cycle. In conclusion, enzymatic activity of PGDH may play an important role in the mechanism involved in luteal resistance to the luteolytic effects of PGF(2alpha). PMID- 11058525 TI - Testis-specific histone H1t gene is hypermethylated in nongerminal cells in the mouse. AB - The testis-specific histone H1t gene is expressed only in pachytene primary spermatocytes during spermatogenesis. There is a correlation between the specific binding of testis nuclear proteins to a rat histone H1t promoter sequence, designated the H1t/TE element, and the onset of transcription in primary spermatocytes. Our laboratory has shown that mice bearing the rat gene with a deletion of the TE promoter element and replacement with a heterologous stuffer DNA fragment fail to express the rat H1t transgene in any tissue. In this study we report that five CpGs located within the H1t proximal promoter, including two CpGs located within the essential TE promoter element, contain unmethylated cytosines in vivo in genomic DNA derived from primary spermatocytes where the H1t gene is expressed. All seven CpGs are hypermethylated in vivo in genomic DNA derived from liver cells where gene expression is repressed. Further, in vitro methylation of an H1t promoter-driven reporter plasmid markedly reduced expression in a transient transfection assay system. These results suggest that cytosine methylation may contribute to the transcriptional silencing of the testis-specific histone H1t gene in nonexpressing tissues such as liver. PMID- 11058526 TI - Ultrastructural evaluation of oocytes during atresia in rat ovarian follicles. AB - Mammalian females are born with a finite number of ovarian oocytes, the vast majority of which ultimately undergo degeneration by atresia. The overall process of ovarian follicular atresia has been morphologically well described only in large antral follicles. Additionally, little attention has been focused on ultrastructural changes in the oocyte. Furthermore, most such morphological studies were performed prior to identification of apoptosis as a mechanism of physiological cell death. Therefore, the purpose of this study was to use electron microscopy to compare the process of atretic oocyte degradation in ovarian follicles of female Fischer 344 rats (38 days old) with ultrastructural characteristics of apoptosis. Examination of ovarian follicles revealed that nucleolar segregation, cytoplasmic or nuclear condensation, apoptotic body formation, and chromatin margination along the nuclear membrane are never observed in atretic oocytes during the degenerative process. Instead, early morphological changes in atretic oocytes include retraction of granulosa cell- and oocyte-derived microvilli and condensation of mitochondria and loss of cristae. These occurrences coincide with initiation of granulosa cell apoptosis. After most granulosa cells are lost, more severe changes occur, including segmentation of the oocyte and cytoplasmic vacuolization as atresia progresses. Thus, these results suggest that, during atresia, oocytes are removed by physiological oocyte cell death, a method that does not involve classically described apoptosis. PMID- 11058527 TI - Periovulatory changes in the local release of vasoactive peptides, prostaglandin f(2alpha), and steroid hormones from bovine mature follicles in vivo. AB - We previously proposed that an endothelin-angiotensin-atrial natriuretic peptide system may contribute to inducing ovulation of mature bovine follicles by modulating follicular secretion of steroids and prostaglandins (PGs). Thus, this study aimed to determine the real-time changes in the local release of angiotensin II (Ang II), endothelin (ET), atrial natriuretic peptide (ANP), PGF(2alpha), and steroid hormones from bovine mature follicles during the periovulatory period in vivo. Seven cows were treated for superovulation using FSH and PGF(2alpha) injections. Two dialysis capillary membranes per follicle were surgically implanted into the theca layer of mature follicles and connected to a microdialysis system (MDS). Fractions of the perfusate were collected from Day -1 (Day 0 = LH surge) to Day 3. Five out of seven treated cows were normally ovulated, and the newly formed corpora lutea were observed at the end of the experiment. In these five ovulated cows, the release of estradiol, androstenedione, and progesterone in the theca layer increased (P < 0.05) synchronously with the LH surge. Acute increases in PGF(2alpha) and Ang II concentrations in the ovarian venous plasma (OVP) were observed at 24-48 h after the peak of the LH surge, when multiple ovulations were expected to occur. The follicular Ang II release was low during the pre-LH surge period and rose (P < 0.05) at the beginning of the increase in the LH surge. On the other hand, ET-1 release dropped (P < 0.05) when plasma LH started to increase. However, no clear changes in ANP concentration in the MDS perfusate and plasma were observed. The above local changes in Ang II, PGF(2alpha), as well as steroid hormones were not observed in cows (n = 2) that did not show an LH surge and ovulation. The present results demonstrate for the first time the local release of Ang II, ET-1, and ANP from the bovine mature follicle in real-time in vivo and show that Ang II and PGF(2alpha) concentrations in the OVP acutely increase around the time of ovulation. The overall results support the concept of a local functional ET-Ang ANP system in the bovine mature follicle that may be involved in the ovulatory process. PMID- 11058528 TI - Gestational changes in uterine L-type calcium channel function and expression in guinea pig. AB - Pregnancy can influence both the resting membrane potential and the ion channel composition of the uterine myometrium. Calcium flux is essential for excitation contraction coupling in pregnant uterus. The uterine L-type calcium channel is an important component in mediating calcium flux and is purported to play a role in parturition. This study was undertaken to characterize gestational changes in 1) the uterine contractile response to the L-type calcium channel agonist, Bay K 8644; 2) the mRNA expression of channel subunits by semiquantitative reverse transcriptase-polymerase chain reaction; and 3) estimate channel protein levels by measuring (3)H-isradipine binding at the dihydropyridine binding site of the alpha(1c) subunit utilizing saturation binding methods. Sensitivity to Bay K 8644 increases beginning at 0.8 of gestation and persists through term. The change in sensitivity is coincident with an increased mRNA expression of the alpha(1c) and beta(2) subunits but with the least detectable amounts of isradipine binding. The expressed alpha(1c) transcript represents a novel structural variant with a 118 amino acid deletion in the III-IV linker and repeats IVS1-S3 of the protein sequence. The guinea pig uterine L-type calcium channel activity is highly regulated through gestation, but the regulation of mRNA expression may be different from regulation of protein levels, estimated by isradipine binding. The up-regulation of function, alpha(1c) subunit mRNA expression, and isradipine binding at term gestation are consistent with a role for this ion channel in parturition. PMID- 11058529 TI - Activation of protein kinase calpha in the lysophosphatidic acid-induced bovine sperm acrosome reaction and phospholipase D1 regulation. AB - Protein kinase C (PKC) has been implicated in the sperm acrosome reaction. In the present study, we demonstrate induction of the acrosome reaction and activation of sperm PKCalpha by lysophosphatidic acid (LPA), which is known to induce signal transduction cascades in many cell types via binding to specific cell-surface receptors. Under conditions by which LPA activates PKCalpha, there is significant stimulation of the acrosome reaction, which is inhibited by PKC inhibitors. Protein kinase Calpha belongs to the Ca(2+)-dependent classical PKC family of isoforms, and indeed we show that its activation depends upon the presence of Ca(2+) in the incubation medium. Protein kinase Calpha is a known regulator of phospholipase D (PLD). We investigated the possible regulatory relationships between PKCalpha and PLD1. Using specific antibodies against PLD1, we demonstrate for the first time its presence in bovine sperm. Furthermore, PLD1 coimmunoprecipitates with PKCalpha and the PKCalpha-PLD1 complex decomposes after treatment of the cells with LPA or 12-O:-tetradecanoyl phorbol-13-acetate, resulting in the translocation of PKCalpha to the plasma membrane and translocation of PLD1 to the particulate fraction. A possible bilateral regulation of PKCalpha and PLD1 activation during the sperm acrosome reaction is suggested. PMID- 11058530 TI - Regulation of Fas antigen (Fas, CD95)-mediated apoptosis of bovine granulosa cells by serum and growth factors. AB - Our previous studies have shown that bovine granulosa cells cultured in basal media supplemented with 5% fetal bovine serum (BM-FBS) are resistant to apoptosis induced by recombinant Fas ligand (FasL) unless pretreated with interferon-gamma (IFN). Experiments were conducted to test the hypothesis that serum and growth factors alter the susceptibility of granulosa cells to FasL-induced apoptosis. Granulosa cells were cultured in BM-FBS, BM containing insulin, transferrin, selenium, and BSA (BM-ITS), and in BM-ITS supplemented with insulin-like growth factor-I (IGF). Cells were susceptible to FasL-induced killing in BM-ITS (27% killing) but were resistant in BM-FBS and in BM-ITS containing IGF (P < 0.05 vs. killing in BM-ITS). Exposure of phosphatidylserine residues on the outer cell membrane, an early marker of apoptosis, was stimulated by FasL and prevented in the presence of IGF. Neutralization of IGF activity in serum with IGF binding protein 3 reduced the protective effect of FBS on FasL-induced killing (P < 0.05), suggesting that IGF is an inhibitory component in FBS. Cotreatment with IFN overcame the inhibitory effects of serum and IGF on FasL-induced killing (31% and 29% killing, respectively, P > 0.05), but IFN did not potentiate killing of cells cultured in BM-ITS. IFN increased expression of Fas antigen (Fas, the receptor for FasL) mRNA five- to sevenfold (P: < 0. 05) and increased immunostaining for Fas protein similarly in all types of media. Addition of the growth factors epidermal growth factor or basic fibroblast growth factor to BM ITS also inhibited FasL-induced killing (P < 0.05), whereas keratinocyte growth factor, transforming growth factor, platelet-derived growth factor, FSH, and LH had no effect. In summary, FasL-induced killing is inhibited by FBS and certain growth factors. IFN increased expression of Fas similarly in all types of media but was required for FasL-induced killing only in BM containing FBS or IGF. Therefore, modulation of responsiveness to FasL-induced apoptosis by growth factors and IFN is not directly related to the level of Fas expression. PMID- 11058531 TI - Inducible nitric oxide synthase in the rat testis: evidence for potential roles in both normal function and inflammation-mediated infertility. AB - In vitro data have indicated that nitric oxide (NO) inhibits Leydig cell testosterone production, suggesting that NO may play a role in the suppression of steroidogenesis and spermatogenic function during inflammation. Consequently, we investigated expression of the inflammation-inducible isoform of NO synthase (iNOS) in the inflamed adult rat testis and the ability of a broad-spectrum inhibitor of NO production, L-nitro-L-arginine methyl ester, to prevent Leydig cell dysfunction during inflammation. Unexpectedly, immunohistochemical and mRNA data established that iNOS is expressed constitutively in Leydig cells and in a stage-specific manner in Sertoli, peritubular, and spermatogenic cells in the normal testis. Expression was increased in a dose-dependent manner in all these cell types during lipopolysaccharide (LPS)-induced inflammation. In noninflamed testes, treatment with the NO synthase inhibitor reduced testicular interstitial fluid formation and testosterone production without any effect on serum LH levels. Administration of the inhibitor did not prevent the suppression of testicular interstitial fluid and testosterone production that occurs within 6 h after LPS treatment. Collectively, these data indicate a novel role for iNOS in autocrine or paracrine regulation of the testicular vasculature, Leydig cell steroidogenesis, and spermatogenesis in the normal testis. The data suggest that increased NO is not the major cause of acute Leydig cell dysfunction in the LPS treated inflammation model, although a role for NO in this process cannot be excluded, particularly at other time points. Moreover, up-regulation of iNOS may contribute to the seminiferous epithelium damage caused by LPS-induced inflammation. PMID- 11058532 TI - Cryobiology of rat embryos I: determination of zygote membrane permeability coefficients for water and cryoprotectants, their activation energies, and the development of improved cryopreservation methods. AB - New rat models are being developed at an exponential rate, making improved methods to cryopreserve rat embryos extremely important. However, cryopreservation of rat embryos has proven to be difficult and expensive. In this study, a series of experiments was performed to characterize the fundamental cryobiology of rat fertilized 1-cell embryos (zygotes) and to investigate the effects of different cryoprotective agents (CPAs) and two different plunging temperatures (T(p)) on post-thaw survival of embryos from three genetic backgrounds. In the initial experiments, information on the fundamental cryobiology of rat zygotes was determined, including 1) the hydraulic conductivity in the presence of CPAs (L(p)), 2) the cryoprotectant permeability (P(CPA)), 3) the reflection coefficient (sigma), and 4) the activation energies for these parameters. P(CPA) values were determined for the CPAs, ethylene glycol (EG), dimethyl sulfoxide (DMSO), and propylene glycol (PG). Using this information, a cryopreservation method was developed and the cryosurvival and fetal development of Sprague-Dawley zygotes cryopreserved in either EG, DMSO, or PG and plunged at either -30 or -80 degrees C, were assessed. The highest fetal developmental rates were obtained using a T(p) of -30 degrees C and EG (61.2% +/- 2.4%), which was not different (P > 0.05) from nonfrozen control zygotes (54.6% +/- 3.0%). PMID- 11058533 TI - Cryobiology of rat embryos II: A theoretical model for the development of interrupted slow freezing procedures. AB - Current mammalian embryo cryopreservation protocols typically employ an interrupted slow freezing (ISF) procedure. In general, ISF consists of initial slow cooling, which raises the extracellular solute concentration, and results in cell dehydration. Permeating cryoprotective agents (CPAs), such as dimethyl sulfoxide (DMSO), are typically included in the medium to protect the cells against high solute concentrations. As this ISF procedure continues, slow cooling is terminated at an intermediate temperature (T(p)), followed by plunging into liquid nitrogen (LN(2)). If the slow cooling step allowed a critical concentration ([CPA](c)) of CPA to be reached within the cell, the CPA will interact with the remaining intracellular water during rapid cooling, resulting in the majority of the intracellular solution becoming vitrified and preventing damaging intracellular ice formation (IIF). This study presents a theoretical model to develop efficient ISF procedures, on the basis of previously developed data for the rat zygote. The model was used to select values of initial CPA concentrations and slow cooling rates (from initial estimated ranges of 0 to 4 molal DMSO and 0 to 2.5 degrees C/min cooling rates) that would allow the intracellular solute concentration to exceed the critical concentration. The optimal combination was then determined from this range based on minimizing the duration of slow cooling. PMID- 11058534 TI - Involvement of apoptosis in the atresia of nonovulatory dominant follicle during the bovine estrous cycle. AB - The present study was designed to 1) investigate whether apoptosis is responsible for the atresia of nonovulatory dominant follicle (DF), 2) to determine if atresia of a nonovulatory DF is associated with alterations in Bcl-2 and Bax expression, 3) to test whether progesterone P(4) has a direct effect on apoptosis in bovine follicles, and 4) to study the pattern of expression of Bcl-2 and Bax in follicles at different developmental stages (small, medium, and large). In experiment 1, 16 cycling cows received a norgestomet ear implant at proestrus (Day 1) for 9 days to mimic the subluteal phase. The cows were assigned either to a control (n = 4) or P(4)-treated groups (n = 12). Injections of P(4) (150 mg, i.m.) were given on Day 3 (n = 4); on Days 3 and 4 (n = 4), and on Days 3, 4, and 5 (n = 4) of the implant period. Controls received injections of corn oil on Days 3, 4, and 5. Unilateral ovariectomy was performed on Days 4, 5, and 6 to recover DFs from cows that had been treated with P(4) for 24, 48, and 72 h, respectively. DFs in the control group were collected on Day 6. The onset of atresia of DFs was assessed morphologically by ultrasound to determine DF diameters, histologically by light microscopic inspection of tissue sections, and functionally by quantification of follicular fluid steroid hormone levels. Apoptosis was detected by DNA analysis and in situ TUNEL labeling. Expression of Bcl-2 and Bax proteins was examined by Western blot analysis. The earliest signs of atresia were detected 24 h after P(4) injection as evidenced by decreased diameter, degeneration and detachment of granulosa cells (GCs) from the basal lamina, and a dramatically reduced ratio of estrogen to P(4). Electrophoretic analysis of DNA extracted from DFs of cows treated with P(4) for 24 h revealed a distinct ladder pattern of DNA fragments. In contrast, this pattern was not obvious in DFs from control cows. Similar results were also obtained from TUNEL analysis of DFs. Furthermore, both Bcl-2 and Bax were found to be present in all DFs; however, the ratio of Bcl-2 and Bax protein levels was significantly reduced by 24 h of P(4) treatment compared with DFs from the control group (P < 0.05). Experiment 2 investigated the direct effect of P(4) (4 ng/ml) on apoptosis of cultured GCs using ovaries obtained from a local slaughterhouse. In addition, the pattern of expressions of Bcl-2 and Bax in follicles at different developmental stages (small, medium, and large) was studied. No increase in apoptotic DNA fragments was detected in GCs treated with P(4). The ratio of Bcl-2 and Bax protein levels was variable in small follicles; however, Bax protein level was always relatively higher than that of Bcl-2 in medium and large follicles. In conclusion, our study suggests that apoptosis is the mechanism that underlies the atresia of nonovulatory DFs that develops during the luteal phase of bovine estrous cycle. PMID- 11058535 TI - Altered hormonal responsiveness of proliferation and apoptosis during myometrial maturation and the development of uterine leiomyomas in the rat. AB - Uterine leiomyomas are responsive to the ovarian steroids, estrogen and progesterone; however, a mechanistic understanding of the role of these hormones in the development of this common gynecologic lesion remains to be elucidated. We have used the Eker rat uterine leiomyoma model to investigate how ovarian hormones regulate or promote the growth of these tumors. Proliferative and apoptotic rates were quantitated in normal uterine tissues and leiomyomas in response to endogenous ovarian steroids. In 2- to 4-mo-old animals, cell proliferation in the normal uterus corresponded with high serum levels of steroid hormones during the estrous cycle, and apoptosis occurred in the rat uterus in all cell types following sharp, cyclical declines in serum hormone levels. It is interesting that the responsiveness of uterine mesenchymal cells changed between 4 and 6 mo of age, with significant decreases in both proliferative and apoptotic rates observed in myometrial and stromal cells of cycling animals. Leiomyomas displayed much higher levels of proliferation than did age-matched myometrium; however, their apoptotic index was significantly decreased in comparison with normal myometrium. This disregulation between proliferative and apoptotic responses, which were tightly regulated during ovarian cycling in the normal myometrium, may contribute to the disruption of tissue homeostasis and underlie neoplastic growth of these tumors. PMID- 11058536 TI - Estrogen receptors alpha and beta in the female reproductive tract of the rat during the estrous cycle. AB - The action of steroid hormones is primarily mediated via a process that involves hormone binding to specific receptors in target cells, which leads to transcriptional activation of steroid-responsive genes and, subsequently, to a modification of cellular responses. The aim of the present study was to obtain information about the dynamics of the two types of estrogen receptors (ERs), alpha and beta, by comparing their concentration and distribution in the reproductive tract of the rat during the estrous cycle. Twenty-four 55- to 60-day old female Sprague-Dawley rats were used. The stage of estrous cycle was determined by vaginal smear. ERalpha was the dominating subtype in uterus, oviduct, and cervix/vagina, with the distribution varying in stroma and epithelium during the estrous cycle. A low level of ERalpha mRNA was observed in ovarian stromal cells, with some scattered positive cells found among granulosa cells. ERbeta expression was observed in the different compartments of uterus and cervix/vagina, but cyclic variation during the estrous cycle was less evident than that of ERalpha. Only a few scattered cells that contained ERbeta mRNA were observed in oviduct. ERbeta mRNA was highly expressed in granulosa cells of developing follicles, with a weaker hybridization signal in new corpora lutea. Immunohistochemistry showed that protein levels of ERalpha and ERbeta have distinct specificity for tissues and cell types, similar to their respective levels of mRNA, as assessed by in situ hybridization. The precise physiological function and importance of ERbeta is still unclear. The relative physiological and pathological function of each ER subtype in the female reproductive tract remains to be further evaluated. PMID- 11058537 TI - Clusterin gene in rat sertoli cells is regulated by a core-enhancer element. AB - Clusterin is a ubiquitous glycoprotein that is promiscuously expressed at a low basal level but can be highly induced by a variety of stress conditions. In contrast, in some secretory cells associated with tissue-fluid interfaces such as the Sertoli cells in the testis, clusterin demonstrates high constitutive expression. In this study, we address the mechanisms that regulate the constitutive expression of the clusterin gene by using primary cultures of immature rat Sertoli cells. We have identified a region of the rat clusterin gene promoter that activated transcription only in Sertoli cells and that mapped between positions -426 and -311. Sequence analysis of this region revealed a high concentration of potential regulatory elements. Using gel-shift assays combined with hydroxyl radical footprinting, we identified the elements recognized by the Sertoli cell nuclear factors. Comparison of the interactions with this region of the nuclear factors from different cell types demonstrated that recognition of the core-enhancer element is specific for the Sertoli cells, and in vitro, the core region was recognized by the transcription factor CBF. Transient transfections showed that a core enhancer is responsible for more than a half of the total promoter activity and is an essential element for the cell-specific activity of the Sertoli-specific region. In addition to the core enhancer, tandem Sp1 sites are also required for maximal activity of this region. PMID- 11058538 TI - Estradiol increases multiunit electrical activity in the A15 area of ewes exposed to inhibitory photoperiods. AB - Seasonal anestrus in ewes results from an increase in response to the negative feedback action of estradiol (E(2)). This increase in the inhibitory effects of E(2) is controlled by photoperiod and appears to be mediated, in part, by dopaminergic neurons in the retrochiasmatic area of the hypothalamus (A15 group). This study was designed to test the hypothesis that E(2) increases multiunit electrical activity (MUA) in the A15 during inhibitory long days. MUA was monitored in the retrochiasmatic area of 14 ovariectomized ewes from 4 h before to 24 h after insertion of an E(2)-containing implant subcutaneously. In six of these ewes, MUA activity was also monitored before and after insertion of blank implants. Three of the 14 ewes were excluded from analysis because E(2) failed to inhibit LH. When MUA was recorded within the A15, E(2) produced a gradual increase in MUA that was sustained for 24 h. Blank implants failed to increase MUA in the A15 area, and E(2) did not alter MUA if recording electrodes were outside the A15. These data demonstrate that E(2) increases MUA in the A15 region of ewes and are consistent with the hypothesis that these neurons mediate E(2) negative feedback during long photoperiods. PMID- 11058539 TI - Identification of potassium-dependent and -independent components of the apoptotic machinery in mouse ovarian germ cells and granulosa cells. AB - Recent studies with thymocytes have suggested a critical role for intracellular potassium in the regulation of apoptosis. In this study, we examined the pathways of K(+) regulation during ovarian cell death. In initial studies, fluorographic analysis demonstrated a significant loss of K(+) during apoptosis stimulated by doxorubicin in oocytes and trophic hormone deprivation in granulosa cells. In oocytes, suppression of potassium efflux by potassium-enriched medium prevented condensation, budding, and fragmentation, although it did not block DNA degradation, suggesting the existence of potassium-independent nucleases in oocytes. Culture of granulosa cells in potassium-enriched medium inhibited internucleosomal DNA cleavage, although high-molecular weight DNA cleavage was apparent, suggesting that the nuclease or nucleases responsible for generating 50 kilobase (kb) fragments in these cells is potassium independent. To address this directly, isolated granulosa cell nuclei were stimulated to autodigest their DNA, and internucleosomal, but not large-fragment, cleavage was completely blocked by 150 mM potassium. We next examined whether the proapoptotic caspases are targets for potassium regulation. In cell-free assays, processing of pro-interleukin 1beta and proteolysis of cellular actin by recombinant caspase-1 and caspase-3, respectively, were suppressed by the presence of 150 mM potassium. Other monovalent ions (NaCl, LiCl) exerted a similar effect in these cell-free assays. Thus, in oocytes and granulosa cells, potassium efflux appears to occur early in the cell death program and may regulate a number of apoptotic events including caspase activity and internucleosomal DNA cleavage. However, there also exist novel potassium-independent pathways in both ovarian germ cells and somatic cells that signal certain apoptotic events, such as large-fragment DNA cleavage. PMID- 11058540 TI - Increased adhesiveness in cultured endometrial-derived cells is related to the absence of moesin expression. AB - Human endometrial epithelial cells (EECs) are nonadhesive for embryos throughout most of the menstrual cycle. During the so-called implantation window, the apical plasma membrane of EECs acquire adhesive properties by undergoing a series of morphological and biochemical changes. The human endometrial-derived epithelial cell line, RL95-2, serves as an in vitro model for receptive uterine epithelium because of its high adhesiveness for trophoblast-derived cells. In contrast, the HEC-1-A cell line, which displays poor adhesive properties for trophoblast cells, is considered to be less receptive. The ezrin, radixin, and moesin protein family members, which are present underneath the apical plasma membrane, potentially act to link the cytoskeleton and membrane proteins. In the present study, we have further investigated the adhesive features in these two unrelated endometrial derived cell lines using an established in vitro model for embryonic adhesion. We have also analyzed the protein pattern and mRNA expression of ezrin and moesin in RL95-2 cells versus HEC-1-A cells. The results demonstrate that RL95-2 cells were indeed more receptive (81% blastocyst adhesion) compared with HEC-1-A cells (46% blastocyst adhesion). An intermediate adhesion rate was found in primary EECs cultured on extracellular matrix gel, thus allowing a partial polarization of these cells (67% blastocyst adhesion). Furthermore, we found that moesin was absent from RL95-2 cells. In contrast, ezrin is expressed in both cell lines, yet it is reduced in adherent RL95-2 cells. Data are in agreement with the hypothesis that uterine receptivity requires down-regulation or absence of moesin, which is a less-polarized actin cytoskeleton. PMID- 11058541 TI - Gene expression of endothelin-1 in the porcine ovary: follicular development. AB - We have investigated which follicular compartment and stage of follicular development are associated with endothelin-1 (ET-1) gene expression in the porcine ovary. The localization of mature ET-1 peptide and of its mRNA was determined by immunohistochemistry and by in situ hybridization. Stage of follicular development associated with ET-1 expression was investigated in terms of follicular class and occurrence of atresia. The latter was investigated by determining the occurrence of DNA fragmentation in apoptotic cells on adjacent sections to those used for ET-1 gene expression. Fifteen ovaries from 10 prepubertal pigs stimulated with gonadotropin were collected; a total of 1050 follicles were examined. Specific ET-1 immunoreactivity was restricted to the ovarian vasculature and to the granulosa cell compartment of antral follicles. The pattern of ET-1 mRNA expression was similar to that found for ET-1 immunoreactivity. Primordial, primary, and most secondary follicles did not express ET-1. The theca cell layer did not express ET-1 regardless of follicle developmental stage. ET-1 expression occurred with a significantly greater probability (P < 0.001 by the likelihood ratio test) in the granulosa cell compartment of antral follicles than in any other follicle class. Furthermore, in antral follicles, ET-1 expression occurred with a greater likelihood in large antral follicles than in small antral follicles (P < 0.001 by the likelihood ratio test). In small antral follicles, only 16.8% expressed ET-1; in contrast, 66.7% of large antral follicles exhibited ET-1 expression. It is interesting that in follicles in which ovulation had already occurred, intense ET-1 expression was found only in the prominent developing vasculature, the other cells present in the luteinized follicle did not display any ET-1 expression. The pattern of ET-1 gene expression observed in this study would be in agreement with our previous suggestion of a plausible physiological role for ET-1 in preventing premature progesterone production by granulosa cells of an antral follicle. The occurrence of atresia and expression of ET-1 in the same follicle was rare. Small and large antral follicles constituted 5.1% and 5.6%, respectively, of the examined follicles in this category. The majority of atretic follicles did not express ET 1 and, conversely, follicles that expressed ET-1 were not atretic. To the best of our knowledge, this is the first report in which large, nonatretic follicles are clearly identified as the population of follicles expressing ET-1. The results of this study delineate the follicular developmental stage and the compartment of when and where ET-1 may be physiologically meaningful. PMID- 11058542 TI - Fluctuations in rat testicular interstitial oxygen tensions are linked to testicular vasomotion: persistence after repair of torsion. AB - Testicular microvascular blood flow is known to exhibit vasomotion, which has been shown to be significantly altered in the short term following the repair of testicular torsion. This loss of vasomotion may ultimately be responsible for the loss of spermatogenesis observed after testicular torsion in rats. In the present study, testicular vasomotion and interstitial oxygen tensions were simultaneously measured prior to, during, and at various time points after repair of testicular torsion in the rat. Testicular torsion was induced by a 720 degrees rotation of the testis for 1 h. Laser-Doppler flowmetry and an oxygen electrode were used to simultaneously measure vasomotion and interstitial oxygen tensions (PO(2)), respectively. Pretorsion control testes had a mean blood flow of 16.3 +/- 1.3 perfusion units (PU) and displayed vasomotion with a cycle frequency of 12 +/- 0.2 cycles per minute and a mean amplitude of 4.2 +/- 0.3 PU. Mean testicular interstitial PO(2) was 12.5 +/- 2.6 mm Hg, which displayed a cyclical variation of 11.9 +/- 0.4 cycles per minute with a mean amplitude of 2.8 +/- 0.8 mm Hg. During the torsion period, both mean blood flow and interstitial PO(2) decreased to approximately zero. Upon detorsion, mean microvascular blood flow and mean interstitial PO(2) values returned to values that were not significantly different from pretorsion values within 30 min; however, vasomotion and PO(2) cycling did not return, even after 24 h. It was 7 days after the repair of torsion before a regular pattern of vasomotion and PO(2) cycling returned. These results demonstrate for the first time a correlation between testicular vasomotion and interstitial PO(2) cycling, and this correlation persists after the repair of testicular torsion. PMID- 11058543 TI - Polarized release of matrix metalloproteinase-2 and -9 from cultured human placental syncytiotrophoblasts. AB - The large increase in placental surface area and fetal villous vascular development in the third trimester of pregnancy requires degradation and reformation of the placental basal lamina. Degradation is carried out by matrix metalloproteinases (MMPs) secreted by adjacent cells. Although the gelatinases, MMP-2 and MMP-9, which are released by extravillous cytotrophoblasts (CTs) are believed to play crucial roles in early placental expansion, neither has been reported in third trimester villous trophoblasts nor has appropriate (basolateral) release of any MMP by the highly polarized syncytiotrophoblast (ST) been demonstrated. We demonstrated villous trophoblast expression of both MMP-2 and MMP-9 by in situ immunohistochemistry and by Western blot analysis and zymography of lysates and culture supernatants of highly purified villous CTs. We also found that epidermal growth factor (EGF)-stimulated CT differentiation into ST and stimulation by the phorbol diester, PMA, both increase MMP-9 secretion. The direction of MMP release was determined with confluent cultures of ST on porous membranes. We found that >90% of MMP-2 and MMP-9 were released from the basolateral surface. We conclude that villous STs express and release gelatinases from their basolateral surfaces in a regulated manner and suggest that such polarized release may be important to villous tissue remodeling. PMID- 11058544 TI - Selection of peptides targeting the human sperm surface using random peptide phage display identify ligands homologous to ZP3. AB - Analysis of the surface architecture of human spermatozoa is a necessary step in the development of new approaches to contraception and resolving the causes of human infertility. In this study we have utilized phage display technology to identify peptides that bind with high affinity to the surface of human spermatozoa. Fifteen- and twelve-mer random peptide phage display libraries were screened against paraformaldehyde-fixed spermatozoa and a number of sperm-binding peptides were identified. One peptide, M6, displayed a high level of affinity for the sperm surface and showed sequence homology with a dominant human ZP3 epitope (hZP 25-33). This peptide bound preferentially to the equatorial and post acrosomal domains of the sperm head and exhibited contraceptive activity by virtue of its capacity to impair the fusion of acrosome-reacted spermatozoa with the vitelline membrane of the oocyte. A similar form of contraceptive activity was also observed within an unrelated peptide, K6, derived from screening the 12 mer library. These results indicate that phage display technology is a powerful tool for developing reagents capable of targeting the human sperm surface, providing insights into the composition of this structure and the identity of targets susceptible to contraceptive attack and pathological disruption. PMID- 11058545 TI - Cimetidine (Tagamet) is a reproductive toxicant in male rats affecting peritubular cells. AB - Cimetidine (Tagamet) is a potent histaminic H2-receptor antagonist, extensively prescribed for ulcers and now available without prescription. Cimetidine is a known testicular toxicant, but its mechanism of action remains uncertain. Rats were treated i.p. with cimetidine either at 50 mg/kg or 250 mg/kg body weight for 59 days. Accessory sex organ weights, but not testis weight, were significantly reduced in the high dose treated groups. FSH levels were significantly elevated in both treated groups, but testosterone levels were unchanged. A high degree of variability characterized testis histology, with most tubules appearing normal and some tubules (15-17%) partially lacking or devoid of germ cells. Morphometry showed that although seminiferous tubule volume was not significantly changed, the volume of peritubular tissue was reduced in the high dose group. There was extensive duplication of the basal lamina, lamina densa in both apparently normal spermatogenic tubules and severely damaged tubules. Apoptotic peritubular myoid cells were also found. TUNEL labeling confirmed extensive apoptotic cell death in peritubular cells, but revealed apoptosis of vascular smooth muscle. Given that 1) peritubular myoid cell apoptosis occurs in apparently normal tubules, that 2) basal lamina disorders are found, and that 3) peritubular cells are lost from the testis, it is suggested that the primary event in cimetidine-related damage is targeted to testicular smooth muscle cells. This is the first in vivo administered toxicant to be described that targets myoid cells, resulting in abnormal spermatogenesis. PMID- 11058546 TI - Anti-apoptotic action of insulin-like growth factor-I during human preimplantation embryo development. AB - Insulin-like growth factor I (IGF-I) has been shown to increase the proportion of embryos forming blastocysts and the number of inner cell mass cells in human and other mammalian preimplantation embryos. Here we examined whether the increased cell number resulted from increased cell division or decreased cell death. Normally fertilized, Day 2 human embryos of good morphology were cultured to Day 6 in glucose-free Earle's balanced salt solution supplemented with 1 mM glutamine, with (n = 42) and without (n = 45) 1.7 nM IGF-I. Apoptotic cells in Day 6 blastocysts were identified using terminal deoxynucleotidyl dUTP terminal transferase (TUNEL) labeling to detect DNA fragmentation and 4'-6-diamidino-2 phenylindole (DAPI) counterstain to evaluate nuclear morphology. The number of nuclei and extent of DNA and nuclear fragmentation was assessed using laser scanning confocal microscopy. IGF-I significantly increased the proportion of embryos developing to the blastocyst stage from 49% (control) to 74% (+IGF-I) (P < 0.05). IGF-I also significantly decreased the mean proportion of apoptotic nuclei from 16.3 +/- 2.9% (-IGF-I) to 8.7 +/- 1.4% (+IGF-I) (P < 0.05). The total number of cells remained similar between both groups (61.7 +/- 4.6 with IGF-I; 54.5 +/- 5.1 without IGF-I). The increased number of blastocysts combined with reduced cell death suggests that IGF-I is rescuing embryos in vitro which would otherwise arrest and acting as a survival factor during preimplantation human development. PMID- 11058547 TI - Relaxation of myometrium by calcitonin gene-related peptide is independent of nitric oxide synthase activity in mouse uterus. AB - Calcitonin gene-related peptide (CGRP) inhibits myometrial contractile activity. However, the responsiveness of the mouse myometrium to CGRP is dependent on the hormonal and gestational stage. The inhibitory effect of CGRP in the myometrium is prominent during gestation and declines at parturition. The present study was undertaken to examine if nitric oxide (NO) production by nitric oxide synthase (NOS) isoforms mediates the inhibitory action of CGRP on uterine contractions as has been suggested earlier. Transgenic mice deficient in either of the three major NOS isoforms: endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS (nNOS) were used. Isometric force measurements on myometrial strips obtained from NOS-deficient mice were carried out and the inhibitory capacity of CGRP was monitored. CGRP inhibited KCl-induced contractions of the myometrial strips obtained from eNOS(-/-), iNOS(-/-), and nNOS(-/-) mice with equal efficiency as in wild-type animals. Additionally, NOS protein expression in the mouse uterus during gestation and during the estrous cycle was examined by means of Western immunoblot analysis. No correlation between NOS expression and inhibitory activity of CGRP was evident. The results suggest that the inhibitory action of CGRP in the mouse uterus is independent of the activity of these NOS isoforms. PMID- 11058548 TI - Identification of a hamster epididymal region-specific secretory glycoprotein that binds nonviable spermatozoa. AB - Even though the epididymis produces an environment promoting sperm maturation and viability, some sperm do not survive transit through the epididymal tubule. Mechanisms that segregate the epididymal epithelium and/or the viable sperm population from degenerating spermatozoa are poorly understood. We report here the identification and characterization of HEP64, a 64-kDa glycoprotein secreted by principal cells of the corpus and proximal cauda epididymidis of the hamster that specifically binds to and coats dead/dying spermatozoa. The HEP64 monomer contains approximately 12 kDa carbohydrate and, following chemical deglycosylation, migrates as a approximately 52-kDa polypeptide. Both soluble (luminal fluid) and sperm-associated HEP64 are assembled into disulfide-linked high molecular weight oligomers that migrate as a doublet band of 260/280 kDa by nonreducing SDS-PAGE. In the epididymal lumen, HEP64 is concentrated into focal accumulations containing aggregates of structurally abnormal or degenerating spermatozoa, and examination of sperm suspensions reveals that HEP64 forms a shroudlike coating surrounding abnormal spermatozoa. The HEP64 glycoprotein firmly binds degenerating spermatozoa and is not released by either nonionic detergent or high salt extraction. Electron microscopic immunocytochemistry demonstrates that HEP64 localized to an amorphous coating surrounding the abnormal spermatozoa. The potential mechanisms by which this epididymal secretory protein binds dead spermatozoa as well as its possible functions in the sperm storage function of the cauda epididymidis are discussed. PMID- 11058549 TI - Zona reaction in porcine oocytes fertilized in vivo and in vitro as seen with scanning electron microscopy. AB - Morphological changes in zona pellucidae (ZP) isolated from in vitro-matured (IVM) and ovulated porcine oocytes were compared before or after fertilization in vitro and in vivo, respectively, by using scanning electron microscopy (SEM). The ZP of some ovulated or IVM oocytes and in vivo- or in vitro-fertilized (IVF) zygotes were equally split into two halves while immersed in an enzyme-inhibitor solution, using a surgical blade. After washing, intact and ZP halves were fixed in 1% glutaraldehyde solution in 0.1 M cacodylate buffer, processed, and examined using SEM. The outer surface of ZP in ovulated oocytes had a mesh-like structure. The outer morphology in IVM oocytes was more smooth although the mesh-like structure was still visible at high magnification. In in vivo zygotes and IVM-IVF zygotes, this lysed, mesh-like structure was more obvious. The inner surface of ZP had some small depressions (orifices). The mean number of orifices per 100 micrometer(2) of ZP surface was larger in IVM oocytes as compared to ovulated ones. The number of orifices per 100 micrometer(2) decreased in IVM-IVF zygotes as compared to IVM oocytes; whereas, in vivo zygotes did not differ from ovulated oocytes. The mean diameter of intact ZP as well as their mean thickness was greater in ovulated oocytes than IVM oocytes. The mean thickness of the ZP was larger in ovulated oocytes than IVM ones. The ZP thickness was larger in zygotes than in in vivo oocytes, whereas that of IVM-IVF zygotes did not differ from that of IVM oocytes. These results indicate that the morphology of ZP and the ZP reaction at sperm penetration appears to be much different between IVM oocytes and ovulated ones. PMID- 11058550 TI - Effect of oxytocin receptor blockade on rat myometrial responsiveness to prostaglandin f(2)(alpha). AB - In the present study we have shown that the genetic expression of prostaglandin (PG)F(2alpha) receptor (R) and cyclooxygenase (COX)-2 increases in laboring rat myometrium. This finding was associated with a relatively weak contractile in vitro response (E:(max)) of isolated uterine strips when challenged with PGF(2alpha). Five days postpartum PGF(2alpha)-R mRNA values exceeded those during labor while COX-2 mRNA was reduced to preparturient values. Maximal contractility of isolated strips stimulated with PGF(2alpha) at this time was enhanced and E:C(50) decreased. Oxytocin treatment of estrogen-primed nonpregnant rats down regulated uterine contractile responsiveness to PGF(2alpha), leaving mRNA values for this receptor unchanged, whereas oxytocin receptor blockade with atosiban (an oxytocin receptor antagonist) left E:(max) unaltered. In contrast, atosiban treatment of pregnant rats resulted in a 2.5-fold increase in E:(max) and a considerably reduced EC(50) during labor when compared to untreated delivering rats. The increased contractile ability was associated with a threefold increase in PGF(2alpha)-R mRNA production, indicating that the regulation by atosiban of the PGF(2alpha)-induced response is exerted at the genetic level. Based on the present data we suggest that 1) PGF(2alpha)-R stimulation may not primarily exert a contracting role in the normally delivering myometrium, and 2) the presence of the PGF(2alpha)-R system in rat myometrium may explain the apparent functional redundancy of the oxytocinergic system during the process of birth in animals lacking oxytocin or where the oxytocin receptor is blocked. In this context PGF(2alpha) receptor stimulation may, in the absence of oxytocin receptor stimulation, exert the contractile forces needed for proper propulsion of the fetus. PMID- 11058551 TI - Changes in turkey semen lipids during liquid in vitro storage. AB - The changes in lipid composition of spermatozoa and seminal plasma and changes in motility, viability, and morphological integrity of spermatozoa were measured in turkey semen diluted in Beltsville poultry semen extender and stored for 48 h (4 degrees C). The total phospholipid content of spermatozoa decreased during storage, while no quantitative decrease was observed in seminal plasma. More precisely, significant decreases in phosphatidylcholine, and to a lesser extent in sphingomyeline, phosphatidylserine, and phosphatidylinositol were observed in spermatozoa. The fatty acid profile of turkey spermatozoa partly reflected diet composition and had a high level of n-9 polyunsaturated fatty acids. Neither fatty acid profile nor free cholesterol were affected by storage. The lipid composition of seminal plasma was quite different from that observed in spermatozoa and was similar to the high density lipoprotein composition of chicken seminal plasma. In vitro storage did not significantly affect lipid classes and only small changes were observed in phospholipid classes of seminal plasma. The motility, viability, and morphological integrity of spermatozoa decreased during storage. These changes in phospholipid content may be explained by membrane phospholipid lysis followed by endogenous metabolism or by a complex combination of lysis, metabolism, and peroxidation. They are likely to affect semen quality and the success of in vitro storage severely. PMID- 11058552 TI - Progesterone receptor-mediated inhibition of apoptosis in granulosa cells isolated from rats treated with human chorionic gonadotropin. AB - Almost all ovarian follicles undergo atresia during follicular development. However, the number of corpora lutea roughly equals the number of preovulatory follicles in the ovary. Because apoptosis is the cellular mechanism behind follicle and luteal cell demise, this suggests a change in apoptosis susceptibility during the periovulatory period. Sex steroids are important regulators of follicular cell survival and apoptosis. The aim of the present work was to study the role of progesterone receptor-mediated effects in the regulation of granulosa cell apoptosis. The levels of internucleosomal DNA fragmentation were evaluated in rat granulosa cells before and after induction of the nuclear progesterone receptor, using hCG treatment to eCG-primed rats to mimic the naturally occurring LH surge. Granulosa cells isolated from hCG-treated rats showed a several-fold increase in the expression of progesterone receptor mRNA and a 47% decrease (P < 0.01) in DNA fragmentation after 24 h incubation in serum free medium compared to granulosa cells isolated from rats treated with eCG only. The effect of hCG treatment in vivo was dose-dependently reversed in vitro by addition of antiprogestins (Org 31710 or RU 486) to the culture medium, demonstrated by increased DNA fragmentation as well as increased caspase-3 activity. Addition of antiprogestins to granulosa cells isolated from immature or eCG-treated rats did not result in increased DNA fragmentation. The results suggest that progesterone receptor-mediated effects are involved in regulating the susceptibility to apoptosis in LH receptor-stimulated preovulatory rat granulosa cells. PMID- 11058553 TI - Molecular pathway of germ cell apoptosis following ischemia/reperfusion of the rat testis. AB - The present study investigates the molecular apoptotic pathway in germ cells following acute ischemia of the rat testis. Rats were subjected to ischemia inducing torsion and testes were harvested after reperfusion. Apoptotic cells were identified with an antibody to single-stranded DNA. Seminiferous tubule RNA was examined by RNase protection assay or by reverse transcriptase-polymerase chain reaction (RT-PCR) for the presence and regulation of apoptotic molecules. Proteins from seminiferous tubules were used for Western blot analysis of cytochrome c. Germ cell apoptosis was maximal at 24 h after repair of torsion. Germ cells in stages II-III of the seminiferous epithelium cycle were the predominant early responders. The RNase protection assays revealed that Bcl-X(L) was the prominent mRNA species. Caspases 1, 2, 3, and Bax mRNA were consistently upregulated; however, the time of upregulation after torsion was variable. The Bcl-X(L) and Bcl-X(S) mRNAs were less consistently upregulated and there was no evidence for upregulation of Fas or Bcl-2. Fas ligand (FasL) was not detected by RNase protection assay, but RT-PCR revealed a significant increase in FasL expression 4 h after the repair of torsion. Western blot analysis for cytochrome c release demonstrated a significant increase 4 h after the repair of torsion. Results suggest that germ cell apoptosis following ischemia/reperfusion of the rat testis is initiated through the mitochondria-associated molecule Bax as well as Fas-FasL interactions. PMID- 11058554 TI - Role of sphingosine in the tumor necrosis factor alpha stimulatory effect on lactate dehydrogenase A expression and activity in porcine Sertoli cells. AB - In this study, the intracellular signaling mechanisms through which TNFalpha increases LDH(A4) activity/expression in primary cultures of porcine testicular Sertoli cells were investigated. Studies were focused on sphingomyelin hydrolysis pathway. Treatment of [(14)C]serine-labeled cells with TNFalpha (15 ng/ml, 0.8 nM) resulted in a transient decrease (approximately 20%) in cellular [(14)C]sphingomyelin and in an increase (approximately 27%) in [(14)C]sphingosine that remained elevated for at least 75 min. In the same experiments, no significant changes were detected in ceramide levels. Exogenous sphingosine stimulated LDH(A4) activity and LDHA expression in a dose-dependent manner (ED(50) = 8 microM of sphingosine). Such an increase in LDHA messenger RNA levels and LDH(A4) activity was detected at 24 h and was maximal after 48 h of treatment. Kinetically, the increase in LDH(A4) activity was similar whether Sertoli cells were treated with sphingosine (12 microM) or with TNFalpha (20 ng/ml). Although sphingosine mimicked the action of TNFalpha on Sertoli cells LDH(A4) activity and expression, the maximal stimulatory effect represented about 30% of TNFalpha maximal activity. Sphingomyelinase, C2 ceramide, sphingosine 1 phosphate, N, N-dimethylsphingosine, and phosphorylcholine had no significant effect on LDHA expression/LDH(A4) activity. Exogenous C2 ceramide increased LDH(A4) activity only in cytokine-treated cells, suggesting its involvement as sphingosine precursor in TNFalpha-stimulated LDH(A4) activity via the sphingomyelin hydrolysis pathway. The LDH(A4) activity stimulated by TNFalpha was decreased by 36.2% by an inhibitor of sphingosine formation, NH4Cl (4 mM), supporting a role of sphingosine in the TNFalpha effect. Moreover, bisindolylmaleimide (100 nM), a protein kinase C (PKC) inhibitor decreased significantly by 28.7% the TNFalpha effect on LDH(A4) activity but had no effect on the stimulating action of sphingosine, suggesting that if PKC is involved in TNFalpha action, the sphingosine effect on LDH(A4) is unrelated to the PKC activity or inhibition. Together, the present data suggest that in primary Sertoli cell cultures, TNFalpha stimulating action on LDHA expression is partly exerted via sphingomyelin hydrolysis pathway, sphingosine being the active metabolite. PMID- 11058555 TI - Evidence that pituitary adenylate cyclase-activating polypeptide is a potent regulator of fetal rat testicular steroidogenesis. AB - Testicular steroidogenesis in the fetal rat is activated before the onset of pituitary gonadotropin secretion. We studied here whether the pituitary adenylate cyclase-activating polypeptide (PACAP) could regulate this early Leydig cell activity. Effects of the two PACAP forms, 27 and 38, were studied on cAMP and testosterone production of dispersed Leydig cells of embryonic Day (E) 18.5. Furthermore, PACAP and PACAP type I receptor mRNA expression were measured by reverse transcription-polymerase chain reaction (RT-PCR), and testicular PACAP concentations by RIA. The two peptides were highly potent stimulators of fetal testes. Doses as low as 10(-18) mol/L of PACAP-27 and 10(-17)-10(-16) mol/L of PACAP-38 significantly stimulated cAMP and testosterone production, with magnitude comparable to that evoked by hCG. These effects were specific for fetal Leydig cells, because PACAP-responsive control cells, including murine Sertoli and granulosa cell lines, only responded to concentrations >/=10(-12) mol/L. By RT-PCR, PACAP and its type I receptor mRNAs were expressed in fetal testis as early as E15.5. By Northern hybridization, PACAP mRNA was first detectable on Day 30 postpartum and increased thereafter. Both forms of PACAP peptides were clearly detectable in E17.5 testes, with decreasing levels thereafter. In conclusion, the steroidogenesis of fetal rat Leydig cells responds to very low concentrations of PACAP, which may be an important physiological regulator of this activity before the onset of pituitary LH secretion. PMID- 11058556 TI - DAZ family proteins exist throughout male germ cell development and transit from nucleus to cytoplasm at meiosis in humans and mice. AB - The human DAZ gene family is expressed in germ cells and consists of a cluster of nearly identical DAZ (deleted in azoospermia) genes on the Y chromosome and an autosomal homolog, DAZL (DAZ-like). Only the autosomal gene is found in mice. Y chromosome deletions that encompass the DAZ genes are a common cause of spermatogenic failure in men, and autosomal homologs of DAZ are essential for testicular germ cell development in mice and Drosophila. Previous studies have reported that mouse DAZL protein is strictly cytoplasmic and that human DAZ protein is restricted to postmeiotic cells. By contrast, we report here that human DAZ and human and mouse DAZL proteins are present in both the nuclei and cytoplasm of fetal gonocytes and in spermatogonial nuclei. The proteins relocate to the cytoplasm during male meiosis. Further observations using human tissues indicate that, unlike DAZ, human DAZL protein persists in spermatids and even spermatozoa. These results, combined with findings in diverse species, suggest that DAZ family proteins function in multiple cellular compartments at multiple points in male germ cell development. They may act during meiosis and much earlier, when spermatogonial stem cell populations are established. PMID- 11058557 TI - Effects of electro-acupuncture on nerve growth factor and ovarian morphology in rats with experimentally induced polycystic ovaries. AB - Despite extensive research on the pathogenesis of polycystic ovary syndrome (PCOS), there is still disagreement on the underlying mechanisms. The rat model for experimentally induced polycystic ovaries (PCO)-produced by a single injection of estradiol valerate-has similarities with human PCOS, and both are associated with hyperactivity in the sympathetic nervous system. Nerve growth factor (NGF) is known to serve as a neurotrophin for both the sympathetic and the sensory nervous systems and to enhance the activity of catecholaminergic and possibly other neuron types. Electro-acupuncture (EA) is known to reduce hyperactivity in the sympathetic nervous system. For these reasons, the model was used in the present study to investigate the effects of EA (12 treatments, approximately 25 min each, over 30 days) by analyzing NGF in the central nervous system and the endocrine organs, including the ovaries. The main findings in the present study were first, that significantly higher concentrations of NGF were found in the ovaries and the adrenal glands in the rats in the PCO model than in the control rats that were only injected with the vehicle (oil or NaCl). Second, that repeated EA treatments in PCO rats resulted in concentrations of NGF in the ovaries that were significantly lower than those in non-EA-treated PCO rats but were within a normal range that did not differ from those in the untreated oil and NaCl control groups. The results in the present study provide support for the theory that EA inhibits hyperactivity in the sympathetic nervous system. PMID- 11058558 TI - Prostaglandin f(2alpha) induces distinct physiological responses in porcine corpora lutea after acquisition of luteolytic capacity. AB - This study examines differences in intracellular responses to cloprostenol, a prostaglandin (PG)F(2alpha) analog, in porcine corpora lutea (CL) before (Day 9 of estrous cycle) and after (Day 17 of pseudopregnancy) acquisition of luteolytic capacity. Pigs on Day 9 or Day 17 were treated with saline or 500 microgram cloprostenol, and CL were collected 10 h (experiment I) or 0.5 h (experiment III) after treatment. Some CL were cut into small pieces and cultured to measure progesterone and PGF(2alpha) secretion. In experiment I, progesterone remained high and PGF(2alpha) low in luteal incubations from either Day 9 or Day 17 saline treated pigs. Cloprostenol increased PGF(2alpha) production 465% and decreased progesterone production 87% only from Day 17 luteal tissue. Cloprostenol induced prostaglandin G/H synthase (PGHS)-2 mRNA (0.5 h) and protein (10 h) in both groups. In cell culture, cloprostenol or phorbol 12, 13-didecanoate (PDD) (protein kinase C activator), induced PGHS-2 mRNA in luteal cells from both groups. However, acute cloprostenol treatment (10 min) decreased progesterone production and increased PGF(2alpha) production only from Day 17 luteal cells. Thus, PGF(2alpha) production is induced by cloprostenol in porcine CL with luteolytic capacity (Day 17) but not in CL without luteolytic capacity (Day 9). However, this change in PGF(2alpha) production is not explained by a difference in induction of PGHS-2 mRNA or protein. PMID- 11058559 TI - Expression of regulator of G-protein signaling protein-2 gene in the rat ovary at the time of ovulation. AB - The ovulatory process in mammals begins when an endogenous surge in LH circulates to the ovary and couples with receptors in the plasma membranes of granulosa cells in mature ovarian follicles. This study provides evidence that the ovulatory stimulus includes induction of the gene for regulator of G-protein signaling protein-2 (RGS2). Immature Wistar rats were primed with 10 IU eCG s.c., and 48 h later the 12-h ovulatory process was initiated by 10 IU hCG (a homolog of LH) s.c. Ovarian RNA was extracted at 0, 2, 4, 8, 12, and 24 h after injecting the animals with hCG. The RNA extracts were used for reverse transcription polymerase chain reaction differential display to detect gene expression in the stimulated ovarian tissue. Two of the amplified cDNAs that were upregulated within 2 h after the ovaries had been stimulated by hCG were homologous to segments of the mouse gene for RGS2. In situ hybridization indicated that the RGS2 mRNA was expressed in the granulosa layer of mature follicles. In conclusion, the gene for RGS2, which is known to regulate membrane signaling pathways, is transcribed in ovarian follicles in response to an ovulatory dose of gonadotropin. PMID- 11058560 TI - Cocaine- and amphetamine-regulated transcript peptide in the rat epididymis: an immunohistochemical and electrophysiological study. AB - Cocaine- and amphetamine-regulated transcript (CART) is a novel family of peptides, of which CART peptide fragments 55-102 and 62-102 are reported to be the endogenous, physiologically active peptides. Immunohistochemical studies with an antiserum directed against the CART peptide fragment 55-102 revealed CART-like immunoreactive (CART-LI) nerve fibers in the rat epididymis. The number was highest in the cauda epididymis and became progressively fewer toward the caput epididymis; the vas deferens exhibited an abundance of CART-LI fibers. Injection of the retrograde tracer Fluorogold (Fluorochrome, Inc., Englewood, CO) to the junction between the vas deferens and cauda epididymis labeled a large number of neurons in the major pelvic ganglion, some of which were CART-positive. Double labeling the ganglion sections with tyrosine hydroxylase (TH) and CART antisera revealed that CART-LI and TH-LI were expressed in two distinct populations of ganglion cells. Some of the TH-LI cells in the ganglia, however, were covered with web-like CART-LI endings. The effects of CART peptide 55-102, referred to herein as CART, on anion secretion in the form of short circuit currents (Isc) were assessed in cultured epithelia. The CART (1 to 5 microM) applied to the basolateral or apical side of the cultured epithelia caused no significant responses on Isc, whereas lys-bradykinin (1 microM) produced a large Isc response in the same preparations. Our results show that CART-LI is present in a population of rat pelvic ganglion cells, which may give rise to CART-LI nerve fibers as observed in the vas deferens and the epididymis. The biological function of CART in the rat epididymis is not known, but it apparently is not involved in ion secretion across the epithelium. PMID- 11058562 TI - Seminal plasma proteins revert the cold-shock damage on ram sperm membrane. AB - Ejaculated ram spermatozoa, freed from seminal plasma by a dextran/swim-up procedure and exposed to cold shock, were incubated with ram seminal plasma proteins and analyzed by fluorescence markers and scanning electron microscopy. Seminal plasma proteins bound to the sperm plasma membrane modified the functional characteristics of damaged spermatozoa, reproducing those of live cells. Scanning electron microscopy showed that the dramatic structural damage induced by cooling reverted after incubation with seminal plasma proteins. Assessment of membrane integrity by fluorescence markers also indicated a restoration of intact-membrane cells. This protein adsorption is a concentration dependent process that induces cell surface restoration in relation to the amount of protein in the incubation medium. Fractionation of ram seminal plasma proteins by exclusion chromatography provided three fractions able to reverse the cold shock effect. Scanning electron microscopy also confirmed the high activity of one fraction, because approximately 50% of cold-shocked sperm plasma membrane surface was restored to its original appearance after incubation. Differences in composition between the three separated fractions mainly resulted from one major band of approximately 20 kDa, which must be responsible for recovering the sperm membrane permeability characteristic of a live cell. PMID- 11058561 TI - Inhibition of reproductive maturation and somatic growth of Fischer 344 rats by photoperiods shorter than L14:D10 and by gradually decreasing photoperiod. AB - Photoperiod is the major regulator of reproduction in temperate-zone mammals. Laboratory rats are generally considered to be nonphotoresponsive, but young male Fischer 344 (F344) rats have a uniquely robust response to short photoperiods of 8 h of light. Rats transferred at weaning from a photoperiod of 16 h to photoperiods of < 14 h of light slowed in both reproductive development and somatic growth rate. Those in photoperiods < 13 h of light underwent the strongest responses. The critical photoperiod of F344 rats can be defined as 13.5 h of light, but photoperiods of 60% of the CD4 cells in naive mice transgenic for a TCR specific for this epitope. The results suggest that the extremely low affinity of the N-terminal peptide for I-A(u) does not limit the MBP1-9-specific T cells from expanding into a sizeable pool of autoreactive T cells. Therefore, the primary immune response to MBP1-9 does not differ quantitatively from previously reported CD4(+) T cell responses to foreign antigens. PMID- 11058576 TI - Invariant chains with the class II binding site replaced by a sequence from influenza virus matrix protein constrain low-affinity sequences to MHC II presentation. AB - Presentation of antigenic peptides by MHC II molecules is required to initiate CD4 T(h) cell responses. Some peptides, however, because of low affinity for MHC II, are not efficiently presented. A segment of the MHC II chaperon molecule, invariant chain (Ii), is known to bind early in biosynthesis with low affinity to the peptide binding groove. Here we have exploited the properties of Ii to manipulate the MHC II-loading pathway and to present low-affinity sequences. We used a deletion mutant of Ii where the promiscuous binding site to MHC II, which is adjacent to the groove binding segment, was deleted. A recombinant Ii (rIi) chimera, derived from this construct, was made in which the class II binding segment was exchanged for wild-type or single amino acid substitution variants of an HLA-DR1-restricted sequence from influenza matrix protein (MAT), which leads to MHC II allotype-specific binding. This rIi was expressed in antigen-presenting cells (APC) and introduced the MAT sequence into the MHC II-processing pathway. As expected, rIiMAT elicited antigen-specific, DR1-restricted T cell cytokine production and proliferation. Significantly, rIiMAT, that binds the HLA-DR4 allele with low affinity, elicited DR4-restricted IL-2 production but not proliferation. In contrast, exogenously provided MAT peptide failed to elicit any responses from DR4-restricted T cells. Compatible results were obtained with a single amino acid substitution variant (MAT(T)), which binds with high affinity to DR4 but low affinity to DR1. We conclude that loading of MHC II with antigenic peptides from endogenously synthesized rIi chimeras allows presentation of low affinity sequences that cannot be presented if provided exogenously as peptides. Ii fusion proteins containing low-affinity antigenic sequences might be useful for vaccination with tumor antigens to overcome deficiencies in antigen presentation. PMID- 11058577 TI - Induction of anti-DNA antibody with DNA-peptide complexes. AB - Spontaneous anti-DNA antibodies in autoimmune mice have the characteristics of antibodies produced by antigen-specific, clonally selective B cell stimulation. The nature of the somatically derived antibody variable region structures recurrent among spontaneous anti-DNA antibodies suggests that DNA or DNA-protein complexes may provide the antigenic stimulus for autoimmune anti-DNA antibody. Previously we have demonstrated that native mammalian DNA in complexes with an immunogenic DNA-binding peptide Fus1 from Trypanosoma cruzi can induce anti-DNA antibody in mice not genetically prone to autoimmune disease. The induced anti DNA has similar specificity, structure and immunopathological function as autoimmune anti-DNA. The present experiments were designed to further characterize the immune response to DNA-peptide complexes. There was considerable variation in the antibody responses of mice from different strains to DNA-Fus1 immunizations. The range was from virtually no response in C57BL/6 mice to most robust responses in NZW mice. The full-length 52 amino acid carboxy-extension protein of ubiquitin (CEP) in T. cruzi (TCEP) protein from which Fus1 was derived functions equally well as an immunogenic carrier for DNA. Anti-DNA responses were generally weak even though anti-Fus1 and anti-TCEP responses were very strong. The results are discussed with respect to the contrasting roles of T cell help and peripheral B cell tolerance in controlling immune and autoimmune antibody responses to DNA. PMID- 11058578 TI - Identification of a novel pre-TCR isoform in which the accessibility of the TCR beta subunit is determined by occupancy of the 'missing' V domain of pre-T alpha. AB - We have identified a novel pre-TCR isoform that is structurally distinct from conventional pre-TCR complexes and whose TCR beta chains are inaccessible to anti TCR beta antibodies. We term this pre-TCR isoform the MB (masked beta)-pre-TCR. Pre-T alpha (pT alpha) subunits of MB-pre-TCR complexes have a larger apparent mol. wt due to extensive modification with O:-linked carbohydrates; however, preventing addition of O-glycans does not restore antibody recognition of the TCR beta subunits of MB-pre-TCR complexes. Importantly, accessibility of TCR beta chains in MB-pre-TCR complexes is restored by filling in the 'missing' variable (V) domain of pT alpha with a V domain from TCR alpha. Moreover, the proportion of pre-TCR complexes in which the TCR beta subunits are accessible to anti-TCR beta antibody varies with the cellular context, suggesting that TCR beta accessibility is controlled by a trans-acting factor. The way in which this factor might control TCR beta accessibility as well as the physiologic relevance of TCR beta masking for pre-TCR function are discussed. PMID- 11058579 TI - Transendothelial migration of 27E10+ human monocytes. AB - The myeloid-related proteins MRP8 (S100A8) and MRP14 (S100A9), two members of the S100 family of calcium-binding proteins, are co-expressed and form a cell-surface and cytoskeleton-associated heterodimer upon calcium mobilization which is recognized by the mAb 27E10. The heterodimer is abundantly expressed in the cytoplasm of granulocytes and a subpopulation of blood monocytes. Previously, we and others demonstrated endothelium-associated MRP8/14 in inflamed tissues in the vicinity of transmigrating leukocytes, suggesting a function of the proteins in this process. Here, we demonstrate that 27E10(+) cells represent a fast-migrating monocyte subpopulation which preferentially utilizes an ICAM-1-dependent mechanism. The following observations imply a function of MRP8/14 in the transmigration process: (i) higher secretion of MRP8/14 from 27E10(+) monocytes compared to 27E10(-) monocytes after interaction with activated endothelium, (ii) higher expression of CD11b on 27E10(+) compared to 27E10(-) monocytes, (iii) up regulation of CD11b on 27E10(-) monocytes in the presence of MRP14 or MRP8/14 heterodimers but not MRP8 and (iv) active participation of MRP14 but not of MRP8 in transmigration as shown by blocking with respective antibodies. We show that the interaction of 27E10(+) monocytes with activated endothelium leads to MRP8/14 release which may account for the high MRP8/14 concentrations in body fluids of patients with acute or chronic inflammatory diseases. Released MRP8/14 may serve a function by enhancing CD11b expression and/or affinity in human monocytes and by participating in the transendothelial migration mechanism. Thus, MRP8/14 substantially contributes to the recruitment of monocytes to an inflammatory site. PMID- 11058580 TI - Immune regulatory effects of central nervous system antigens in culture. AB - Evidence from several different experimental systems suggests that regulatory cells specific for self-antigens exist in the normal immune repertoire, and that these cells are necessary for maintenance of self-tolerance and prevention of autoimmune disease. We attempted to demonstrate the existence of regulatory cells specific for central nervous system (CNS) antigens in normal mice. We tested the effects of myelin basic protein (MBP), glial fibrillary acidic protein (GFAP) and a mixture of soluble brain proteins (SBP) on cultured splenocytes. MBP at 50 microg/ml inhibited antigen-driven proliferation and this suppressive effect could be partially blocked by neutralizing antibodies to transforming growth factor (TGF)-beta. MBP decreased expression of mRNA for the cytokines IL-2 and IFN-gamma, and slightly increased mRNA expression for TGF-beta. These effects did not appear to be mediated by regulatory cells specific for MBP, since MBP also suppressed proliferation in MBP-deficient shiverer mice and the suppressive effect could not be reproduced with selected MBP peptides. SBP at 250 microg/ml also inhibited antigen-driven proliferation, but this effect could not be blocked by neutralizing antibodies against IL-4, IL-10 or TGF-beta. SBP reduced expression of mRNA for IL-2, IL-10 and TGF-beta. These results are more consistent with the presence of a soluble inhibitory factor than with the action of SBP-specific regulatory cells. GFAP had no significant effect on proliferation. These results do not support the existence of regulatory cells specific for CNS antigens. Further investigation into non-antigen-specific mechanisms will be important in defining how autoimmune damage in the CNS is prevented. PMID- 11058581 TI - NKT lymphocyte ontogeny and function are impaired in low antibody-producer Biozzi mice: gene mapping in the interval-specific congenic strains raised for immunomodulatory genes. AB - NKT cells are CD4(+) or CD4(-)CD8(-) CD1d-restricted lymphocytes, characterized by the property to rapidly produce IL-4 and IFN-gamma in response to TCR ligation. This IL-4 burst is lacking in autoimmunity-prone SJL and NOD strains of mice, which suggests an immunoregulatory role for NKT cells. The NKT cell status was thus investigated in the genetically selected high (H) and low (L) antibody producer mice. The results show that (i) the frequency of cells expressing the NKT cell markers is 3- to 4-fold lower in thymus and spleen from L than H mice, (ii) L mice spleen cells did not produce IL-4 following injection of anti-TCR alpha beta antibody, and (iii) L mice thymus and spleen cells failed to produce IL-4 after in vitro stimulation by anti-TCR alpha beta antibody or alpha galactosylceramide, a newly described NKT cell ligand. These parameters were investigated in six interval-specific congenic strains raised for the quantitative trait loci which contain the immunomodulatory genes responsible for the high/low antibody production phenotypes. IL-4 production recovery occurred only in the congenic strain in which the H origin chromosome 4 segment was introgressed on the L background. This finding was not due to increased NKT cell frequency but appeared dependent of antigen-presenting cells in co-culture experiments. This result strongly suggests the presence of gene(s) modulating NKT function on chromosome 4, close to several genes predisposing to autoimmunity. PMID- 11058582 TI - CD4+ T lymphocytes as a primary cellular target for BAT mAb stimulation. AB - BAT is a monoclonal antibody (mAb) produced against membranes of a human Burkitt lymphoma cell line (Daudi) that was selected for its ability to stimulate lymphocyte proliferation. BAT manifests anti-tumor properties in mice bearing a variety of murine tumors. BAT also induced regression of human tumors inoculated into SCID mice that had been engrafted with human lymphocytes. The anti-tumor activity of BAT was related to its immune stimulatory properties. Previous data indicated that T lymphocytes and NK cells mediate in vivo the anti-tumor activity. In order to define the primary target cell for BAT stimulatory activity, the in vitro stimulatory effect of BAT on purified lymphocyte subpopulations was investigated. Human CD4(+), CD8(+) T cells and CD56(+) NK cells were purified and their in vitro response to BAT was investigated. Results indicate that BAT selectively stimulated CD4(+) cells as assessed by proliferation and secretion of IFN-gamma. FACS analysis has also revealed a selective increase in BAT antigen on CD4(+) T cells that were cultured with BAT antibody. The effector cells that mediate BAT-induced tumor eradication may, however, be distinct from those that serve as the primary cellular target of the antibody. Cytokines such as IFN-gamma that are produced by CD4(+) cells may be involved in activation of additional cell types that may be involved in tumor destruction. PMID- 11058583 TI - Rac/Cdc42 and p65PAK regulate the microtubule-destabilizing protein stathmin through phosphorylation at serine 16. AB - We have identified a rapid protein phosphorylation event at residue serine 16 of stathmin using two-dimensional gel electrophoresis coupled to matrix-assisted laser desorption/ionization mass spectrometry in combination with post-source decay analysis, which is induced by the epidermal growth factor receptor. Phosphorylation is specifically mediated by the small GTPases Rac and Cdc42 and their common downstream target, the serine/threonine kinase p65PAK. Both GTPases have previously been shown to regulate the dynamics of actin polymerization. Because stathmin destabilizes microtubules, and this process is inhibited by phosphorylation at residue 16, Rac and Cdc42 can potentially regulate both F actin and microtubule dynamics. PMID- 11058584 TI - Analysis of biological effects and signaling properties of Flt-1 (VEGFR-1) and KDR (VEGFR-2). A reassessment using novel receptor-specific vascular endothelial growth factor mutants. AB - Endothelial cells express two related vascular endothelial growth factor (VEGF) receptor tyrosine kinases, KDR (kinase-insert domain containing receptor, or VEGFR-2) and Flt-1 (fms-like tyrosine kinase, or VEGFR-1). Although considerable experimental evidence links KDR activation to endothelial cell mitogenesis, there is still significant uncertainty concerning the role of individual VEGF receptors for other biological effects such as vascular permeability. VEGF mutants that bind to either KDR or Flt-1 with high selectivity were used to determine which of the two receptors serves to mediate different VEGF functions. In addition to mediating mitogenic signaling, selective KDR activation was sufficient for the activation of intracellular signaling pathways implicated in cell migration. KDR stimulation caused tyrosine phosphorylation of both phosphatidylinositol 3-kinase and phospholipase Cgamma in primary endothelial cells and stimulated cell migration. KDR-selective VEGF was also able to induce angiogenesis in the rat cornea to an extent indistinguishable from wild type VEGF. We also demonstrate that KDR, but not Flt-1, stimulation is responsible for the induction of vascular permeability by VEGF. PMID- 11058585 TI - Characterization of p190RhoGEF, a RhoA-specific guanine nucleotide exchange factor that interacts with microtubules. AB - Rho family GTPases control numerous cellular processes including cytoskeletal reorganization and transcriptional activation. Rho GTPases are activated by guanine nucleotide exchange factors (GEFs) which stimulate the exchange of bound GDP for GTP. We recently isolated a putative GEF, termed p190RhoGEF that binds to RhoA and, when overexpressed in neuronal cells, induces cell rounding and inhibits neurite outgrowth. Here we show that the isolated tandem Dbl homology/pleckstrin homology domain of p190RhoGEF activates RhoA in vitro, but not Rac1 or Cdc42, as determined by GDP release and protein binding assays. In contrast, full-length p190RhoGEF fails to activate RhoA in vitro. When overexpressed in intact cells, however, p190RhoGEF does activate RhoA with subsequent F-actin reorganization and serum response factor-mediated transcription. Immunofluorescence studies show that endogenous p190RhoGEF localizes to distinct RhoA-containing regions at the plasma membrane, to the cytosol and along microtubules. In vitro and in vivo binding experiments show that p190RhoGEF directly interacts with microtubules via its C-terminal region adjacent to the catalytic Dbl homology/pleckstrin homology domain. Our results indicate that p190RhoGEF is a specific activator of RhoA that requires as yet unknown binding partners to unmask its GDP/GTP exchange activity in vivo, and they suggest that p190RhoGEF may provide a link between microtubule dynamics and RhoA signaling. PMID- 11058586 TI - Cloning and characterization of an atypical Type IV P-type ATPase that binds to the RING motif of RUSH transcription factors. AB - RUSH proteins are SWI/SNF-related transcription factors with RING finger signatures near their COOH termini. Long suspected of mediating protein-protein interactions, the RING motif was used to clone a binding partner. The RING finger binding protein (RFBP) is a Type IV P-type ATPase, a putative phospholipid pump, with conserved sequences for two loop segments, an ATP-binding site, a phosphorylation domain, and transmembrane passes potentially involved in substrate binding and translocation. However, RFBP differs from all other Type IV P-type ATPases in three ways. It has only three of four highly conserved NH(2) terminal transmembrane passes, it is located in the inner nuclear membrane, and it binds the RING domain. Topographically the orientation of the adjacent hydrophilic domains and the determinants of transport specificity are altered. As a result, the small, hydrophilic loop extends into the perinuclear space that is contiguous with the lumen of the endoplasmic reticulum. The large, conformationally flexible loop extends into the nucleoplasm to contact euchromatin. Competitive reverse transcriptase-polymerase chain reaction and high performance liquid chromatography analysis revealed that endometrial RFBP mRNA expression is hormonally regulated. The physical association of a hormone dependent RING finger-binding protein with transcriptionally active chromatin supports the speculation that RFBP plays a role in the subnuclear trafficking of transcription factors with RING motifs. PMID- 11058587 TI - Oligomeric state of the colon carcinoma-associated glycoprotein GA733-2 (Ep CAM/EGP40) and its role in GA733-mediated homotypic cell-cell adhesion. AB - The GA733-2 antigen (GA733) is a homotypic calcium-independent cell adhesion molecule (CAM) present in most normal human epithelial cells and gastrointestinal carcinomas. Because oligomerization of some CAMs regulates cell adhesion and signal transduction, the correlation between GA733 oligomeric state and cell-cell adhesion was investigated. Sedimentation equilibrium studies showed that full length (-FL) GA733 exists as dimers and tetramers in solution, whereas the GA733 extracellular domain (-EC) is a monomer. The Kd of GA733-FL is less than 10 nm for the monomer-dimer association, whereas the dimer-tetramer association is about 1000-fold weaker (Kd approximately 10 microm). Chemical cross-linking of purified GA733-FL in solution resulted in a major product corresponding to GA733 dimers, and minor amounts of trimers and tetramers. However, GA733-EC cross linked under the same conditions was consistently a monomer. Chemical cross linking of dissociated colon carcinoma cells produced predominantly GA733 dimers, whereas cross-linking of cells in monolayers yielded some tetramers as well. GA733-FL retained its cell-cell adhesion function as shown by inhibition of cell aggregation, whereas monomeric GA733-EC was inactive. These data show that GA733 exists predominantly as high affinity noncovalent cis-dimers in solution and on dissociated colon carcinoma cells. The lower affinity association of dimers to form tetramers is most likely the head-to-head interaction between GA733 cis dimers on opposing cells that represents its cell-cell adhesion activity. PMID- 11058588 TI - beta 1,3-Galactosyltransferase beta 3Gal-T5 acts on the GlcNAcbeta 1-->3Galbeta 1 ->4GlcNAcbeta 1-->R sugar chains of carcinoembryonic antigen and other N-linked glycoproteins and is down-regulated in colon adenocarcinomas. AB - We attempted to determine whether beta1,3-galactosyltransferase beta3Gal-T5 is involved in the biosynthesis of a specific subset of type 1 chain carbohydrates and expressed in a cancer-associated manner. We transfected Chinese hamster ovary (CHO) cells expressing Fuc-TIII with beta3Gal-T cDNAs and studied the relevant glycoconjugates formed. beta3Gal-T5 directs synthesis of Lewis type 1 antigens in CHO cells more efficiently than beta3Gal-T1, whereas beta3Gal-T2, -T3, and -T4 are almost unable to direct synthesis. In the clone expressing Fuc-TIII and beta3Gal-T5 (CHO-FT-T5), sialyl-Lewis a synthesis is strongly inhibited by swainsonine but not by benzyl-alpha-GalNAc, and sialyl-Lewis x is absent, although it is detected in the clones expressing Fuc-TIII and beta3Gal-T1 (CHO-FT T1) or Fuc-TIII and beta3Gal-T2 (CHO-FT-T2). Endo-beta-galactosidase treatment of N- glycans prepared from clone CHO-FT-T5 releases (+/-NeuAcalpha2-->3)Galbeta1- >3[Fucalpha1-->4]GlcNAcbeta1-->3Gal but not GlcNAcbeta1-->3Gal or type 2 chain oligosaccharides, which are found in CHO-FT-T1 cells. This result indicates that beta3Gal-T5 expression prevents poly-N-acetyllactosamine and sialyl-Lewis x synthesis on N-glycans. Kinetic studies confirm that beta3Gal-T5 prefers acceptors having the GlcNAcbeta1-->3Gal end, including lactotriosylceramide. Competitive reverse transcriptase mediated-polymerase chain reaction shows that the beta3Gal-T5 transcript is expressed in normal colon mucosa but not or poorly in adenocarcinomas. Moreover, recombinant carcinoembryonic antigen purified from a CHO clone expressing Fuc-TIII and beta3Gal-T5 reacts with anti-sialyl-Lewis a and carries type 1 chains on oligosaccharides released by endo-beta galactosidase. We conclude that beta3Gal-T5 down-regulation plays a relevant role in determining the cancer-associated glycosylation pattern of N-glycans. PMID- 11058589 TI - Inhibition of sodium-calcium exchange by ceramide and sphingosine. AB - Na(+)/Ca(2+) exchange activity in Chinese hamster ovary cells expressing the bovine cardiac Na(+)/Ca(2+) exchanger was inhibited by the short chain ceramide analogs N-acetylsphingosine and N-hexanoylsphingosine (5-15 micrometer). The sphingolipids reduced exchange-mediated Ba(2+) influx by 50-70% and also inhibited the Ca(2+) efflux mode of exchange activity. The biologically inactive ceramide analog N-acetylsphinganine had only modest effects on exchange activity. Cells expressing the Delta(241-680) and Delta(680-685) deletion mutants of the Na(+)/Ca(2+) exchanger were not inhibited by ceramide; these mutants show defects in both Na(+)-dependent and Ca(2+)-dependent regulatory behavior. Another mutant, which was defective only in Na(+)-dependent regulation, was as sensitive to ceramide inhibition as the wild-type exchanger. Inhibition of exchange activity by ceramide was time-dependent and was accelerated by depletion of internal Ca(2+) stores. Sphingosine (2.5 micrometer) also inhibited the Ca(2+) influx and efflux modes of exchange activity in cells expressing the wild-type exchanger; sphingosine did not affect Ba(2+) influx in the Delta(241-680) mutant. The effects of the exogenous sphingolipids were reproduced by blocking cellular ceramide utilization pathways, suggesting that exchange activity is inhibited by increased levels of endogenous ceramide and/or sphingosine. We propose that sphingolipids impair Ca(2+)-dependent activation of the exchanger and that in cardiac myocytes, this process serves as a feedback mechanism that links exchange activity to the diastolic concentration of cytosolic Ca(2+). PMID- 11058590 TI - Hyaluronidase induction of a WW domain-containing oxidoreductase that enhances tumor necrosis factor cytotoxicity. AB - To determine how hyaluronidase increases certain cancer cell sensitivity to tumor necrosis factor (TNF) cytotoxicity, we report here the isolation and characterization of a hyaluronidase-induced murine WW domain-containing oxidoreductase (WOX1). WOX1 is composed of two N-terminal WW domains, a nuclear localization sequence, and a C-terminal alcohol dehydrogenase (ADH) domain. WOX1 is mainly located in the mitochondria, and the mitochondrial targeting sequence was mapped within the ADH domain. Induction of mitochondrial permeability transition by TNF, staurosporine, and atractyloside resulted in WOX1 release from mitochondria and subsequent nuclear translocation. TNF-mediated WOX1 nuclear translocation occurred shortly after that of nuclear factor-kappaB nuclear translocation, whereas both were independent events. WOX1 enhanced TNF cytotoxicity in L929 cells via its WW and ADH domains as determined using stable cell transfectants. In parallel with this observation, WOX1 also enhanced TRADD (TNF receptor-associated death domain protein)-mediated cell death in transient expression experiments. Antisense expression of WOX1 raised TNF resistance in L929 cells. Enhancement of TNF cytotoxicity by WOX1 is due, in part, to its significant down-regulation of the apoptosis inhibitors Bcl-2 and Bcl-x(L) (>85%), but up-regulation of pro-apoptotic p53 ( approximately 200%) by the ADH domain. When overexpressed, the ADH domain mediated apoptosis, probably due to modulation of expression of these proteins. The WW domains failed to modulate the expression of these proteins, but sensitized COS-7 cells to TNF killing and mediated apoptosis in various cancer cells independently of caspases. Transient cotransfection of cells with both p53 and WOX1 induced apoptosis in a synergistic manner. WOX1 colocalizes with p53 in the cytosol and binds to the proline-rich region of p53 via its WW domains. Blocking of WOX1 expression by antisense mRNA abolished p53 apoptosis. Thus, WOX1 is a mitochondrial apoptogenic protein and an essential partner of p53 in cell death. PMID- 11058591 TI - The yeast glycerol 3-phosphatases Gpp1p and Gpp2p are required for glycerol biosynthesis and differentially involved in the cellular responses to osmotic, anaerobic, and oxidative stress. AB - We have characterized the strongly homologous GPP1/RHR2 and GPP2/HOR2 genes, encoding isoforms of glycerol 3-phosphatase. Mutants lacking both GPP1 and GPP2 are devoid of glycerol 3-phosphatase activity and produce only a small amount of glycerol, confirming the essential role for this enzyme in glycerol biosynthesis. Overproduction of Gpp1p and Gpp2p did not significantly enhance glycerol production, indicating that glycerol phosphatase is not rate-limiting for glycerol production. Previous studies have shown that expression of both GPP1 and GPP2 is induced under hyperosmotic stress and that induction partially depends on the HOG (high osmolarity glycerol) pathway. We here show that expression of GPP1 is strongly decreased in strains having low protein kinase A activity, although it is still responsive to osmotic stress. The gpp1Delta/gpp2Delta double mutant is hypersensitive to high osmolarity, whereas the single mutants remain unaffected, indicating GPP1 and GPP2 substitute well for each other. Transfer to anaerobic conditions does not affect expression of GPP2, whereas GPP1 is transiently induced, and mutants lacking GPP1 show poor anaerobic growth. All gpp mutants show increased levels of glycerol 3-phosphate, which is especially pronounced when gpp1Delta and gpp1Delta/gpp2Delta mutants are transferred to anaerobic conditions. The addition of acetaldehyde, a strong oxidizer of NADH, leads to decreased glycerol 3-phosphate levels and restored anaerobic growth of the gpp1Delta/gpp2Delta mutant, indicating that the anaerobic accumulation of NADH causes glycerol 3-phosphate to reach growth-inhibiting levels. We also found the gpp1Delta/gpp2Delta mutant is hypersensitive to the superoxide anion generator, paraquat. Consistent with a role for glycerol 3-phosphatase in protection against oxidative stress, expression of GPP2 is induced in the presence of paraquat. This induction was only marginally affected by the general stress-response transcriptional factors Msn2p/4p or protein kinase A activity. We conclude that glycerol metabolism plays multiple roles in yeast adaptation to altered growth conditions, explaining the complex regulation of glycerol biosynthesis genes. PMID- 11058592 TI - Human factor H-related protein 5 (FHR-5). A new complement-associated protein. AB - A novel human plasma protein has been identified as a universal component of complement deposits, when complement is detected immunohistochemically in vivo. The protein is homologous to complement factor H and related proteins and has been designated factor H-related protein 5 (FHR-5). FHR-5 was identified by a monoclonal antibody raised using pathologic human glomerular preparations as the immunogen. FHR-5 was purified by affinity chromatography from complement-lysed erythrocytes, and the peptide sequence was obtained. The cDNA was cloned from a human liver library, and FHR-5 was deduced to be a protein containing 551 amino acids organized into nine short consensus repeat motifs. The short consensus repeats of FHR-5 show homology to Factor H and to other Factor H-related proteins, with some unique features demonstrated. Recombinant FHR-5, expressed in insect cells, was shown to bind C3b in vitro. The strong association of FHR-5 with tissue complement deposits in vivo suggests that this additional member of the Factor H family of proteins has a function in complement regulation. PMID- 11058593 TI - Signal peptidase and oligosaccharyltransferase interact in a sequential and dependent manner within the endoplasmic reticulum. AB - We demonstrate that the signal peptides of prepro-alpha-factor and preinvertase must be cleaved before Asn-X-Ser/Thr acceptor tripeptides located near the signal peptides of these precursors can be efficiently glycosylated within the endoplasmic reticulum of the yeast Saccharomyces cerevisiae. The data support a model whereby the interaction of a signal peptide with the membrane prevents an acceptor tripeptide juxtaposed to the signal peptide from accessing the oligosaccharyltransferase active site until the signal peptide is cleaved. PMID- 11058595 TI - Recruitment of a specific amoeboid myosin I isoform to the plasma membrane in chemotactic Dictyostelium cells. AB - The Dictyostelium class I myosins, MyoA, -B, -C, and -D, participate in plasma membrane-based cellular processes such as pseudopod extension and macropinocytosis. Given the existence of a high affinity membrane-binding site in the C-terminal tail domain of these motor proteins and their localized site of action at the cortical membrane-cytoskeleton, it was of interest to determine whether each myosin I was directly associated with the plasma membrane. The membrane association of a myosin I heavy chain kinase that regulates the activity of one of the class I myosins, MyoD was also examined. Cellular fractionation experiments revealed that the majority of the Dicyostelium MyoA, -B, -C and -D heavy chains and the kinase are cytosolic. However, a small, but significant, fraction (appr. 7. -15%) of each myosin I and the kinase was associated with the plasma membrane. The level of plasma membrane-associated MyoB, but neither that of MyoC nor MyoD, increases up to 2-fold in highly motile, streaming cells. These results indicate that Dictyostelium specifically recruits myoB to the plasma membrane during directed cell migration, consistent with its known role in pseudopod formation. PMID- 11058594 TI - Vascular endothelial growth factor KDR receptor signaling potentiates tumor necrosis factor-induced tissue factor expression in endothelial cells. AB - Vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF alpha) have been shown to synergistically increase tissue factor (TF) expression in endothelial cells; however, the role of the VEGF receptors (KDR, Flt-1, and neuropilin) in this process is unclear. Here we report that VEGF binding to the KDR receptor is necessary and sufficient for the potentiation of TNF-induced TF expression in human umbilical vein endothelial cells. TF expression was evaluated by Western blot analysis and fluorescence-activated cell sorting. In the absence of TNF-alpha, wild-type VEGF- or KDR receptor-selective variants induced an approximate 7-fold increase in total TF expression. Treatment with TNF alone produced an approximate 110-fold increase in total TF expression, whereas coincubation of TNF-alpha with wild-type VEGF- or KDR-selective variants resulted in an approximate 250-fold increase in TF expression. VEGF lacking the heparin binding domain was also able to potentiate TF expression, indicating that heparin sulfate proteoglycan or neuropilin binding is not required for TF up-regulation. Neither placental growth factor nor an Flt-1-selective variant was capable of inducing TF expression in the presence or absence of TNF. Inhibition of protein tyrosine kinase or protein kinase C activity significantly blocked the TNF/VEGF potentiation of TF up-regulation, whereas phorbol 12-myristate 13-acetate, a protein kinase C activator, increased TF expression. These data demonstrate that KDR receptor signaling governs both VEGF-induced TF expression and the potentiation of TNF-induced up-regulation of TF. PMID- 11058596 TI - Importance of the anticodon sequence in the aminoacylation of tRNAs by methionyl tRNA synthetase and by valyl-tRNA synthetase in an Archaebacterium. AB - The mode of recognition of tRNAs by aminoacyl-tRNA synthetases and translation factors is largely unknown in archaebacteria. To study this process, we have cloned the wild type initiator tRNA gene from the moderate halophilic archaebacterium Haloferax volcanii and mutants derived from it into a plasmid capable of expressing the tRNA in these cells. Analysis of tRNAs in vivo show that the initiator tRNA is aminoacylated but is not formylated in H. volcanii. This result provides direct support for the notion that protein synthesis in archaebacteria is initiated with methionine and not with formylmethionine. We have analyzed the effect of two different mutations (CAU-->CUA and CAU-->GAC) in the anticodon sequence of the initiator tRNA on its recognition by the aminoacyl tRNA synthetases in vivo. The CAU-->CUA mutant was not aminoacylated to any significant extent in vivo, suggesting the importance of the anticodon in aminoacylation of tRNA by methionyl-tRNA synthetase. This mutant initiator tRNA can, however, be aminoacylated in vitro by the Escherichia coli glutaminyl-tRNA synthetase, suggesting that the lack of aminoacylation is due to the absence in H. volcanii of a synthetase, which recognizes the mutant tRNA. Archaebacteria lack glutaminyl-tRNA synthetase and utilize a two-step pathway involving glutamyl tRNA synthetase and glutamine amidotransferase to generate glutaminyl-tRNA. The lack of aminoacylation of the mutant tRNA indicates that this mutant tRNA is not a substrate for the H. volcanii glutamyl-tRNA synthetase. The CAU-->GAC anticodon mutant is most likely aminoacylated with valine in vivo. Thus, the anticodon plays an important role in the recognition of tRNA by at least two of the halobacterial aminoacyl-tRNA synthetases. PMID- 11058597 TI - IL-17E, a novel proinflammatory ligand for the IL-17 receptor homolog IL-17Rh1. AB - We report identification of interleukin (IL)-17E, a novel member of the IL-17 family of cytokines. IL-17E is a ligand for the recently identified protein termed EVI27/IL-17BR, which we term IL-17 receptor homolog 1 (IL-17Rh1) in light of the multiple reported ligand-receptor relationships. Murine EVI27 was identified through its location at a common site of retroviral integration in BXH2 murine myeloid leukemias. IL-17Rh1 shows highest level expression in kidney with moderate expression in multiple other organs, whereas IL-17E mRNA was detected at very low levels in several peripheral tissues. IL-17E induces activation of NF-kappaB and stimulates production of the proinflammatory chemokine IL-8. PMID- 11058598 TI - Retinoic acid-mediated activation of the mouse renin enhancer. AB - Previous studies demonstrate that the mouse renin gene is regulated by a complex enhancer of transcription located 2.6 kilobases upstream of the transcription start site which is under both positive and negative influence. We demonstrate herein that a positive regulatory element (Eb) is repeated 10 bp upstream (Ec), and both are required for baseline activity of the enhancer. The Eb and Ec core sequences are identical to the consensus sequence for the nuclear hormone receptor superfamily of transcription factors, and transcriptional activity of constructs containing the enhancer is increased after treatment with retinoic acid. Maximal induction requires both Eb and Ec. Expression of endogenous renin and a renin-promoter controlled transgene in As4.1 cells, and kidney renin mRNA in C57BL/6J mice was induced after retinoid treatment. Gel mobility supershift analysis revealed the binding of RARalpha and RXRalpha to oligonucleotides containing both Eb and Ec. Reverse transcriptase-polymerase chain reaction analysis revealed that As4.1 cells express both receptor isoforms, along with RARgamma, but do not express RARbeta, RXRbeta, or RXRgamma. Co-transfection of an expression vector encoding wild-type RARalpha increased enhancer activity, whereas a dominant negative mutant of RARalpha significantly attenuated retinoic acid-induced activity of the enhancer. These results demonstrate the importance of the Eb and Ec motifs in controlling baseline activity of the renin enhancer, and suggest the potential importance of retinoids in regulating renin expression. PMID- 11058600 TI - Functional role of critical stripe residues in transmembrane span 7 of the serotonin transporter. Effects of Na+, Li+, and methanethiosulfonate reagents. AB - Mutations at critical residue positions in transmembrane span 7 (TM7) of the serotonin transporter affect the Na(+) dependence of transport. It was possible that these residues, which form a stripe along one side of the predicted alpha helix, formed part of a water-filled pore for Na(+). We tested whether cysteine substitutions in TM7 were accessible to hydrophilic, membrane-impermeant methanethiosulfonate (MTS) reagents. Although all five cysteine-containing mutants tested were sensitive to these reagents, noncysteine control mutants at the same positions were in most cases equally sensitive. In all cases, MTS sensitivity could be traced to changes in accessibility of a native cysteine residue in extracellular loop 1, Cys-109. Moreover, none of the TM7 cysteines reacted with the biotinylating reagent MTSEA-biotin when tested in the C109A background. It is thus unlikely that the critical stripe forms part of a water filled pore. Instead, studies of the ion dependence of the reaction between Cys 109 and MTS reagents lead to the conclusion that TM7 is involved in propagating conformational changes caused by ion binding, perhaps as part of the translocation mechanism. The critical stripe residues on TM7 probably represent a close contact region between TM7 and one or more other TMs in the transporter's three-dimensional structure. PMID- 11058599 TI - Executioner caspase-3, -6, and -7 perform distinct, non-redundant roles during the demolition phase of apoptosis. AB - Apoptosis is orchestrated by a family of cysteine proteases known as the caspases. Fourteen mammalian caspases have been identified, three of which (caspase-3, -6, and -7) are thought to coordinate the execution phase of apoptosis by cleaving multiple structural and repair proteins. However, the relative contributions that the "executioner" caspases make to the demolition of the cell remains speculative. Here we have used cell-free extracts immuno depleted of either caspase-3, -6, or -7 to examine the caspase requirements for apoptosis-associated proteolysis of 14 caspase substrates as well as nuclear condensation, chromatin margination, and DNA fragmentation. We show that caspase 3 is the primary executioner caspase in this system, necessary for cytochrome c/dATP-inducible cleavage of fodrin, gelsolin, U1 small nuclear ribonucleoprotein, DNA fragmentation factor 45 (DFF45)/inhibitor of caspase activated DNase (ICAD), receptor-interacting protein (RIP), X-linked inhibitor of apoptosis protein (X-IAP), signal transducer and activator of transcription-1 (STAT1), topoisomerase I, vimentin, Rb, and lamin B but not for cleavage of poly(ADP-ribose) polymerase (PARP) or lamin A. In addition, caspase-3 was also essential for apoptosis-associated chromatin margination, DNA fragmentation, and nuclear collapse in this system. Surprisingly, although caspase-6 and -7 are considered to be important downstream effector caspases, depletion of either caspase had minimal impact on any of the parameters investigated, calling into question their precise role during the execution phase of apoptosis. PMID- 11058601 TI - Defective signaling to Fyn by a T cell antigen receptor lacking the alpha -chain connecting peptide motif. AB - A key role in the communication between the alphabetaTCR and the CD3/zeta complex is played by a specific motif within the connecting peptide domain of the TCR alpha chain (alpha-CPM). T cell hybridomas expressing an alpha-CPM-mutated TCR show a dramatic impairment in antigen-driven interleukin-2 production. This defect can be complemented by a calcium ionophore, indicating that activation of the calcium pathway is impaired. Several lines of evidence implicate Fyn in the regulation of calcium mobilization, at least in part through the activation of phospholipase Cgamma. Here we have investigated the potential involvement of Fyn in the TCR alpha-CPM signaling defect. Using T cell hybridomas expressing either a wild-type TCR or an alpha-CPM mutant, we show that Fyn fails to be activated by the mutant receptor following SEB binding and fails to generate tyrosine phosphorylated Pyk2, a member of the focal adhesion kinase family. This defect correlated with an impairment in phospholipase Cgamma phosphorylation. Production of interlukin-2 and activation of the transcription factor NF-AT in response to triggering of the TCR alpha-CPM mutant with SEB were fully restored in the presence of constitutively active Fyn. Hence the signaling defect generated by the TCR alpha-CPM mutation results at least in part from an impaired coupling of the TCR.CD3 complex to Fyn activation. PMID- 11058602 TI - Sugar binding properties of the two lectin domains of the tandem repeat-type galectin LEC-1 (N32) of Caenorhabditis elegans. Detailed analysis by an improved frontal affinity chromatography method. AB - The 32-kDa galectin (LEC-1 or N32) of the nematode Caenorhabditis elegans is the first example of a tandem repeat-type galectin and is composed of two domains, each of which is homologous to typical vertebrate 14-kDa-type galectins. To elucidate the biological meaning of this unique structure containing two probable sugar binding sites in one molecule, we analyzed in detail the sugar binding properties of the two domains by using a newly improved frontal affinity chromatography system. The whole molecule (LEC-1), the N-terminal lectin domain (Nh), and the C-terminal lectin domain (Ch) were expressed in Escherichia coli, purified, and immobilized on HiTrap gel agarose columns, and the extent of retardation of various sugars by the columns was measured. To raise the sensitivity of the system, we used 35 different fluorescence-labeled oligosaccharides (pyridylaminated (PA) sugars). All immobilized proteins showed affinity for N-acetyllactosamine-containing N-linked complex-type sugar chains, and the binding was stronger for more branched sugars. Ch showed 2-5-fold stronger binding toward all complex-type sugars compared with Nh. Both Nh and Ch preferred Galbeta1-3GlcNAc to Galbeta1-4GlcNAc. Because the Fucalpha1-2Galbeta1 3GlcNAc (H antigen) structure was found to interact with all immobilized protein columns significantly, the K(d) value of pentasaccharide Fucalpha1-2Galbeta1 3GlcNAcbeta1-3Galbeta1-4Glc-PA for each column was determined by analyzing the concentration dependence. Obtained values for immobilized LEC-1, Nh, and Ch were 6.0 x 10(-5), 1.3 x 10(-4), and 6.5 x 10(-5) m, respectively. The most significant difference between Nh and Ch was in their affinity for GalNAcalpha1 3(Fucalpha1-2)Galbeta1-3GlcNAcbeta1-3Galbeta1-4Glc-PA, which contains the blood group A antigen; the K(d) value for immobilized Nh was 4.8 x 10(-5) m, and that for Ch was 8.1 x 10(-4) m. The present results clearly indicate that the two sugar binding sites of LEC-1 have different sugar binding properties. PMID- 11058603 TI - The yeast gene MSC2, a member of the cation diffusion facilitator family, affects the cellular distribution of zinc. AB - The sequence of the yeast gene YDR205W places it within the family of cation diffusion facilitators: membrane proteins that transport transition metals. Deletion of YDR205W was reported to result in an increase in unequal sister chromatid recombination and was named meiotic sister chromatid recombination 2 (MSC2; Thompson, D. A., and Stahl, F. W. (1999) Genetics 153, 621-641). We report here that a msc2 strain shows a phenotype of decreased viability in glycerol ethanol media at 37 degrees C. Associated with decreased growth is an abnormal morphology typified by an increase in size of both cells and vacuoles. Addition of extracellular Zn2+ completely suppresses the morphological changes and partially suppresses the growth defect. Regardless of the concentration of Zn2+ in the media, the msc2 strain had a higher Zn2+ content than wild type cells. Zinquin staining also revealed that msc2 had a marked increase in fluorescence compared with the wild type, again reflecting an increase in intracellular Zn2+. The deletion strain accumulated excess Zn2+ in nuclei-enriched membrane fractions, and when grown at 37 degrees C in glycerol-ethanol media, it showed a decreased expression of Zn2+-regulated genes. The expression of genes regulated by either Fe2+ or Cu2+ was not affected. An epitope-tagged Msc2p was localized to the endoplasmic reticulum/nucleus. These results suggest that Msc2p affects the cellular distribution of zinc and, in particular, the zinc content of nuclei. PMID- 11058604 TI - Interaction of the transcription factors USF1, USF2, and alpha -Pal/Nrf-1 with the FMR1 promoter. Implications for Fragile X mental retardation syndrome. AB - Hypermethylation of the FMR1 promoter reduces its transcriptional activity, resulting in the mental retardation and macroorchidism characteristic of Fragile X syndrome. How exactly methylation causes transcriptional silencing is not known but is relevant if current attempts to reactivate the gene are to be successful. Understanding the effect of methylation requires a better understanding of the factors responsible for FMR1 gene expression. To this end we have identified five evolutionarily conserved transcription factor binding sites in this promoter and shown that four of them are important for transcriptional activity in neuronally derived cells. We have also shown that USF1, USF2, and alpha-Pal/Nrf-1 are the major transcription factors that bind the promoter in brain and testis extracts and suggest that elevated levels of these factors account in part for elevated FMR1 expression in these organs. We also show that methylation abolishes alpha Pal/Nrf-1 binding to the promoter and affects binding of USF1 and USF2 to a lesser degree. Methylation may therefore inhibit FMR1 transcription not only by recruiting histone deacetylases but also by blocking transcription factor binding. This suggests that for efficient reactivation of the FMR1 promoter, significant demethylation must occur and that current approaches to gene reactivation using histone deacetylase inhibitors alone may therefore have limited effect. PMID- 11058605 TI - Human Nod1 confers responsiveness to bacterial lipopolysaccharides. AB - The immune response to microbial pathogens is initiated by recognition of specific pathogen components by host cells both at the cell surface and in the cytosol. While the response triggered by pathogen products at the surface of immune cells is well characterized, that initiated in the cytosol is poorly understood. Nod1 is a member of a growing family of intracellular proteins with structural homology to apoptosis regulators Apaf-1/Ced-4 and a class of plant disease-resistant gene products. Here we show that bacterial lipopolysaccharides, but not other pathogen components tested, induced TLR4- and MyD88-independent NF kappaB activation in human embryonic kidney 293T cells expressing trace amounts of Nod1. Nod2, another Nod family member, also conferred responsiveness to bacterial components but with a response pattern different from that observed with Nod1. As it was reported for plant disease-resistant R proteins, the leucine rich repeats of Nod1 and Nod2 were required for lipopolysaccharide-induced NF kappaB activation. A lipopolysaccharide binding activity could be specifically coimmunopurified with Nod1 from cytosolic extracts. These observations suggest that Nod1 and Nod2 are mammalian counterparts of plant disease-resistant gene products that may function as cytosolic receptors for pathogen components derived from invading bacteria. PMID- 11058606 TI - Role of perspective and other uncertainties in cost-effectiveness assessments in advanced prostate cancer. PMID- 11058607 TI - Colorectal cancer: does it matter if you eat your fruits and vegetables? PMID- 11058608 TI - U.S. Presidential candidates define policies on cancer. PMID- 11058609 TI - Stat bite: Importance of health care issues in the 2000 presidential election. PMID- 11058610 TI - Laser capture microdissection comes into mainstream use. PMID- 11058611 TI - Genetic testing intensifies research on psychological impact of cancer. PMID- 11058612 TI - What is the role of bacteria in cancer carcinogenesis? PMID- 11058613 TI - Silicone breast implants not linked to breast cancer risk. PMID- 11058614 TI - National Cancer Institute uses breast cancer stamp proceeds to fund research. PMID- 11058615 TI - Metastasis-suppressor genes: a review and perspective on an emerging field. AB - Metastasis is the most lethal attribute of a cancer. There is a critical need for markers that will distinguish accurately those histologic lesions and disseminated cells with a high probability of causing clinically important metastatic disease from those that will remain indolent. While the development of new diagnostic markers of metastasis was the initial motivation for many studies, the biologic approach used to identify metastasis-suppressor genes has provided surprising insights into the in vivo mechanisms regulating the formation of metastases. This review and perspective describes the evolving view of the mechanisms that regulate metastasis and the importance of metastasis-suppressor genes in this process. The known metastasis-suppressor proteins or genes and the microcell-mediated chromosomal transfer strategy used to identify many of them are reviewed. New evidence for the role of these metastasis-suppressor proteins or genes in regulating the growth of disseminated cancer cells at the secondary site, the potential for the identification of novel therapeutic targets, and the multidisciplinary approach needed to translate this information into clinical tools for the treatment of metastatic disease are discussed. PMID- 11058616 TI - Cost-effectiveness of androgen suppression therapies in advanced prostate cancer. AB - BACKGROUND: The costs and side effects of several antiandrogen therapies for advanced prostate cancer differ substantially. We estimated the cost effectiveness of antiandrogen therapies for advanced prostate cancer. METHODS: We performed a cost-effectiveness analysis using a Markov model based on a formal meta-analysis and literature review. The base case was assumed to be a 65-year old man with a clinically evident, local recurrence of prostate cancer. The model used a societal perspective and a time horizon of 20 years. Six androgen suppression strategies were evaluated: diethylstilbestrol (DES), orchiectomy, a nonsteroidal antiandrogen (NSAA), a luteinizing hormone-releasing hormone (LHRH) agonist, and combinations of an NSAA with an LHRH agonist or orchiectomy. Outcome measures were survival, quality-adjusted life years (QALYs), lifetime costs, and incremental cost-effectiveness ratios. RESULTS: DES, the least expensive therapy, had a discounted lifetime cost of $3600 and the lowest quality-adjusted survival, 4.6 QALYs. At a cost of $7000, orchiectomy was associated with 5.1 QALYs, resulting in an incremental cost-effectiveness ratio of $7500/QALY relative to DES. All other strategies-LHRH agonists, NSAA, and both combined androgen blockade strategies-had higher costs and lower quality-adjusted survival than orchiectomy. These results were sensitive to the quality of life associated with orchiectomy and the efficacy of combined androgen blockade, and they changed little when prostate-specific antigen results were used to guide therapy. Under a wide range of other assumptions, the cost-effectiveness of orchiectomy relative to DES was consistently less than $20 000/QALY. Androgen suppression therapies were most cost-effective if initiated after patients became symptomatic from prostate metastases. CONCLUSIONS: For men who accept it, orchiectomy is likely to be the most cost-effective androgen suppression strategy. Combined androgen blockade is the least economically attractive option, yielding small health benefits at high relative costs. PMID- 11058617 TI - Prospective study of fruit and vegetable consumption and incidence of colon and rectal cancers. AB - BACKGROUND: Frequent consumption of fruit and vegetables has been associated with a reduced risk of colorectal cancer in many observational studies. METHODS: We prospectively investigated the association between fruit and vegetable consumption and the incidence of colon and rectal cancers in two large cohorts: the Nurses' Health Study (88 764 women) and the Health Professionals' Follow-up Study (47 325 men). Diet was assessed and cumulatively updated in 1980, 1984, 1986, and 1990 among women and in 1986 and 1990 among men. The incidence of cancer of the colon and rectum was ascertained up to June or January of 1996, respectively. Relative risk (RR) estimates were calculated with the use of pooled logistic regression models accounting for various potential confounders. All statistical tests were two-sided. RESULTS: With a follow-up including 1 743 645 person-years and 937 cases of colon cancer, we found little association of colon cancer incidence with fruit and vegetable consumption. For women and men combined, a difference in fruit and vegetable consumption of one additional serving per day was associated with a covariate-adjusted RR of 1.02 (95% confidence interval [CI] = 0.98-1.05). A difference in vegetable consumption of one additional serving per day was associated with an RR of 1.03 (95% CI = 0.97 1.09). Similar results were obtained for women and men considered separately. A difference in fruit consumption of one additional serving per day was associated with a covariate-adjusted RR for colon cancer of 0.96 (95% CI = 0.89-1.03) among women and 1. 08 (95% CI = 1.00-1.16) among men. For rectal cancer (total, 244 cases), a difference in fruit and vegetable consumption of one additional serving per day was associated with an RR of 1.02 (95% CI = 0.95-1.09) in men and women combined. None of these associations was modified by vitamin supplement use or smoking habits. CONCLUSIONS: Although fruits and vegetables may confer protection against some chronic diseases, their frequent consumption does not appear to confer protection from colon or rectal cancer. PMID- 11058618 TI - Prospective study of serum selenium levels and incident esophageal and gastric cancers. AB - BACKGROUND: From March 1986 through May 1991, we conducted a randomized nutritional intervention trial, the General Population Trial, in Linxian, China, a region with epidemic rates of squamous esophageal and adenomatous gastric cardia cancers. We found that participants who received selenium, beta-carotene, and vitamin E had significantly lower cancer mortality rates than those who did not. In the current study, we examined the relationship between selenium levels measured in pretrial (1985) sera from participants and the subsequent risk of developing squamous esophageal, gastric cardia, and gastric non-cardia cancers during the trial. METHODS: This study was designed and analyzed in accord with a stratified case-cohort sampling scheme, with the six strata defined by sex and three age categories. We measured serum selenium levels in 590 case subjects with esophageal cancer, 402 with gastric cardia cancers, and 87 with gastric non cardia cancers as well as in 1062 control subjects. Relative risks (RRs), absolute risks, and population attributable risk for cancers were estimated on the basis of the Cox proportional hazards models. All statistical tests are two sided. RESULTS: We found highly significant inverse associations of serum selenium levels with the incidence of esophageal (P: for trend <10(-4)) and gastric cardia (P: for trend <10(-6)) cancers. The RR and 95% confidence interval (CI) for comparison of highest to lowest quartile of serum selenium was 0.56 (95% CI = 0.44-0.71) for esophageal cancer and 0.47 (95% CI = 0.33-0.65) for gastric cardia cancer. The population proportion of these cancers that is attributable to low selenium levels was 26.4% (95% CI = 14.45-38.36). We found no evidence for a gradient of serum selenium associated with incidence of gastric non-cardia cancer (P: for trend =.96), with an RR of 1.07 (95% CI = 0.55-2.08) for the highest to lowest quartile of serum selenium. CONCLUSIONS: Our study supports findings from previous prospective studies and randomized trials that variations in selenium levels affect the incidence of certain cancers. In the United States, where intervention trials of selenium are in the planning stages, consideration should be given to including populations at high risk for squamous esophageal and gastric cardia cancers. PMID- 11058619 TI - Repair of tobacco carcinogen-induced DNA adducts and lung cancer risk: a molecular epidemiologic study. AB - BACKGROUND: Only a fraction of cigarette smokers develop lung cancer, suggesting that people differ in their susceptibility to this disease. We investigated whether differences in DNA repair capacity (DRC) for repairing tobacco carcinogen induced DNA damage are associated with differential susceptibility to lung cancer. METHODS: From August 1, 1995, through April 30, 1999, we conducted a hospital-based, case-control study of 316 newly diagnosed lung cancer patients and 316 cancer-free control subjects matched on age, sex, and smoking status. DRC was measured in cultured lymphocytes with the use of the host-cell reactivation assay with a reporter gene damaged by a known activated tobacco carcinogen, benzo[a]pyrene diol epoxide. Statistical tests were two-sided. RESULTS: Overall, lower DRC was observed in case patients than in control subjects (P:<.001) and was associated with a greater than twofold increased risk of lung cancer. Compared with the highest DRC quartile in the control subjects and after adjustment for age, sex, pack-years of smoking, family history of cancer, and other covariates, reduced DRC was associated with increased risk of lung cancer in a dose-dependent fashion (odds ratio [OR] = 1.8 with 95% confidence interval [CI] = 1.1-3.1, OR = 2.0 with 95% CI = 1.2-3.4, and OR = 4. 3 with 95% CI = 2.6 7.2 for the second, third, and fourth quartiles, respectively; P:(trend)<.001). Case patients who were younger at diagnosis (<60 years old), female, or lighter smokers or who reported a family history of cancer exhibited the lowest DRC and the highest lung cancer risk among their subgroups, suggesting that these subgroups may be especially susceptible to lung cancer. CONCLUSION: The results provide evidence that low DRC is associated with increased risk of lung cancer. The findings from this hospital-based, case-control study should be validated in prospective studies. PMID- 11058620 TI - Minimization of heterocyclic amines and thermal inactivation of Escherichia coli in fried ground beef. AB - BACKGROUND: Heterocyclic amine carcinogens are formed during the cooking of a number of foods, especially well-done meats. Lower temperatures and shorter cooking times can minimize the formation of these carcinogens, yet a major food safety concern is that pathogens in the meat must be thermally inactivated. This study investigated cooking techniques that minimize heterocyclic amine formation while simultaneously destroying contaminating bacteria. METHODS: Ground beef patties were inoculated with Escherichia coli K12 bacteria and fried to internal temperatures ranging from 35 degrees C to 70 degrees C in a skillet preheated to 160 degrees C, 180 degrees C, or 200 degrees C. Each patty was then analyzed for four common heterocyclic amines and for surviving bacteria. Additionally, the frequency of turning of the beef patty during cooking was varied (a single turn or multiple turns), length of time required for each patty to reach 70 degrees C was recorded, and heterocyclic amine levels were determined. An additional pan temperature of 250 degrees C was tested for its effect on heterocyclic amine formation but not on bacterial killing. Statistical tests were two-sided. RESULTS: Colony-forming bacteria were reduced by five orders of magnitude at internal temperatures greater than 60 degrees C, regardless of cooking method, and were completely inactivated at 70 degrees C. For patties turned just once, heterocyclic amine levels increased as the cooking temperatures increased. However, levels of heterocyclic amines were statistically significantly lower with turning every minute. For each pan temperature, patties reached 70 degrees C internal temperature sooner when they were turned every minute than when they were turned just once during cooking. CONCLUSION: Lowering the pan temperature and turning the patties frequently can greatly reduce the formation of heterocyclic amines and can simultaneously achieve bacterial inactivation with little or no increase in cooking time, ensuring a product that is safe for human consumption. PMID- 11058621 TI - Molecular analysis of urine sediment for follow-up of urinary tract cancers. PMID- 11058622 TI - Report on the first meeting of the International Collaborative Group on Hereditary Gastric Cancer. PMID- 11058623 TI - Evolution of diagnostic neuroradiology from 1904 to 1999. AB - Neuroradiology began in the early 1900s soon after Roentgen discovered x rays, with the use of skull radiographs to evaluate brain tumors. This was followed by the development of ventriculography in 1918, pneumoencephalography in 1919, and arteriography in 1927. In the beginning, air studies were the primary modality, but this technique was supplanted by angiography in the 1950s and 1960s. The first full-time neuroradiologist in the United States was Cornelius G. Dyke at the New York Neurological Institute in 1930. Neuroradiology took a firm hold as a specialty in the early 1960s when Dr Juan M. Taveras brought together fourteen neuroradiologists from the United States and Canada to establish the nucleus of what was to become the American Society of Neuroradiology, or ASNR. This society's initial goals were to perform research and to advance knowledge within the specialty. Neuroradiologists initially were able to diagnose vascular disease, infections, tumors, trauma, and alterations in cerebrospinal fluid flow, because the brain structure was invisible. Neuroradiology was forever changed with computed tomography (CT) because the brain structure became visible. Soon thereafter, magnetic resonance (MR) imaging was developed, and it not only provided anatomic but also made possible vascular and physiologic functional imaging. PMID- 11058624 TI - Anatomic extent of disease: a critical variable in reports of diagnostic accuracy. PMID- 11058625 TI - Low back pain. AB - The communication between radiologists and their surgical colleagues is particularly important in the setting of back pain. This common disorder often does not have a definable cause, even when the imaging findings are abnormal. A shared understanding of the various causes of back pain, the appropriate terminology, and the needs of the surgeon is vital to proper patient treatment. Unfortunately, little standardization in the terminology for and management of back pain syndromes exists. This article elucidates the approaches to problems of back pain used in one clinical setting. PMID- 11058626 TI - Diffusion-weighted MR imaging of the brain. AB - Diffusion-weighted magnetic resonance (MR) imaging provides image contrast that is different from that provided by conventional MR techniques. It is particularly sensitive for detection of acute ischemic stroke and differentiation of acute stroke from other processes that manifest with sudden neurologic deficits. Diffusion-weighted MR imaging also provides adjunctive information for other cerebral diseases including neoplasms, intracranial infections, traumatic brain injury, and demyelinating processes. Because stroke is common and in the differential diagnosis of most acute neurologic events, diffusion-weighted MR imaging should be considered an essential sequence, and its use in most brain MR studies is recommended. PMID- 11058627 TI - Sialolithiasis and salivary ductal stenosis: diagnostic accuracy of MR sialography with a three-dimensional extended-phase conjugate-symmetry rapid spin echo sequence. AB - PURPOSE: To evaluate the accuracy of magnetic resonance (MR) sialography in detecting salivary glandular calculi and ductal stenoses. MATERIALS AND METHODS: In a prospective study, 64 salivary glands in 61 consecutive patients with acute or recurrent parotid or submandibular glandular swelling were examined by using three-dimensional (3D) extended-phase conjugate-symmetry rapid spin-echo (EXPRESS) MR imaging. Transverse and sagittal-oblique source images and maximum intensity projection images were obtained. All MR images were analyzed independently by two radiologists, without knowledge of the final diagnosis. The reference standard was conventional sialography, ultrasonography (US), and sialendoscopy with or without surgery in 31 glands and was conventional sialography and US in 33 glands. RESULTS: Final diagnoses included sialolithiasis (n = 23), sialolithiasis and stenosis (n = 9), stenosis without lithiasis (n = 11), early Sjogren syndrome without ductal stenosis (n = 2), ductal displacement (n = 3), and normal salivary glands (n = 16). The sensitivity, specificity, and positive and negative predictive values of MR sialography to detect calculi were 91%, 94%-97%, 93%-97%, and 91%, respectively. False-negative readings occurred due to calculi with a diameter of 2-3 mm in nondilated salivary ducts. Ductal stenosis was assessed, with a sensitivity of 100%, specificity of 93%-98%, positive predictive value of 87%-95%, and negative predictive value of 100%. Interobserver agreement was very good (kappa = 0.85-0.97). CONCLUSION: MR sialography with 3D EXPRESS imaging enables reliable prediction of salivary gland calculi and stenoses. PMID- 11058628 TI - The fat C2 sign. PMID- 11058629 TI - Can noninvasive imaging accurately depict intracranial aneurysms? A systematic review. AB - PURPOSE: To perform a systematic review to determine the accuracy of computed tomographic (CT) angiography, magnetic resonance (MR) angiography, and transcranial Doppler ultrasonography (US) in depicting intracranial aneurysms. MATERIALS AND METHODS: A 1988-1998 literature search for studies with 10 or more subjects in which noninvasive imaging was compared with angiography was undertaken. Studies meeting initial criteria were evaluated by using intrinsically weighted standardized assessment to determine suitability for inclusion. Studies scoring greater than 50% were included. RESULTS: Of 103 studies that met initial criteria, 38 scored greater than 50%. CT angiography and MR angiography had accuracies per aneurysm of 89% (95% CI: 87%, 91%) and 90% (95% CI: 87%, 92%), respectively. For US, data were scanty and accuracy was lower, although the CIs overlapped those of CT angiography and MR angiography. Sensitivity was greater for detection of aneurysms larger than 3 mm than for detection of aneurysms 3 mm or smaller-for CT angiography, 96% (95% CI: 94%, 98%) versus 61% (95% CI: 51%, 70%), and for MR angiography, 94% (95% CI: 90%, 97%) versus 38% (95% CI: 25%, 53%). Diagnostic accuracy was similar for anterior and posterior circulation aneurysms. CONCLUSION: CT angiography and MR angiography depicted aneurysms with an accuracy of about 90%. Most studies were performed in populations with high aneurysm prevalence, which may have introduced bias toward noninvasive examinations. PMID- 11058630 TI - Preeclampsia-eclampsia: clinical and neuroradiographic correlates and insights into the pathogenesis of hypertensive encephalopathy. AB - PURPOSE: To investigate the clinical parameters that are associated with the development of brain edema of hypertensive encephalopathy in patients with preeclampsia-eclampsia. MATERIALS AND METHODS: Twenty-eight patients with preeclampsia-eclampsia and neurologic symptoms underwent magnetic resonance (MR) imaging. Clinical parameters recorded at the time of MR imaging included serum electrolytes and various indices of hematologic, renal, and hepatic function. Several data were available 1 week prior to the development of neurologic symptoms in 11 patients. Univariate analysis and multivariate logistic regression analyses were performed to study possible associations between these parameters and brain edema at MR imaging. RESULTS: The 20 patients with brain edema at MR imaging had a significantly greater incidence of abnormal red blood cell morphology (14 [82%] of 17 patients vs two [25%] of eight, P: <.005) and higher levels of lactic dehydrogenase (LDH) (339 U/L +/- 65 [SD] vs 258 U/L +/- 65, P: =.007) than the eight with normal MR imaging findings; multivariate logistic regression analysis showed a strong association with red blood cell morphology only. Moreover, LDH levels were elevated before the development of neurologic abnormalities (P: <.05). Blood pressures were not significantly different between groups at any time. CONCLUSION: Brain edema at MR imaging in patients with preeclampsia-eclampsia was associated with abnormalities in endothelial damage markers and not with hypertension level. PMID- 11058631 TI - Malignant gliomas: MR imaging spectrum of radiation therapy- and chemotherapy induced necrosis of the brain after treatment. AB - PURPOSE: To describe both the common and less frequently encountered magnetic resonance (MR) imaging features of radiation therapy- and chemotherapy-induced brain injury, with particular emphasis on radiation necrosis. MATERIALS AND METHODS: A cohort of 148 adult patients underwent surgical resection of malignant brain (glial) tumors and were subsequently entered into a research protocol that consisted of accelerated radiation therapy with carboplatin followed by chemotherapy with procarbazine, lomustine, and vincristine. Patients typically underwent sequential MR imaging at 6-8-week intervals during the 1st year and at 3-6-month intervals during subsequent years. In all patients, histopathologic confirmation of lesion composition was performed by board-certified neuropathologists. RESULTS: The patients exhibited different types of MR imaging detected abnormalities of the brain: pure radiation necrosis in 20 patients, a mixture of predominantly radiation necrosis with limited recurrent and/or residual tumor (less than 20% of resected tissue) in 16 patients, radiation necrosis of the cranial nerves and/or their pathways in two patients, radiation induced enhancement of the white matter in 52 patients, and radiation-induced enhancement of the cortex in nine patients. CONCLUSION: The frequent diagnostic dilemma of recurrent neoplasm versus radiation necrosis is addressed in this study through a description of the varying spatial and temporal patterns of radiation necrosis at MR imaging. PMID- 11058632 TI - Monitoring of tumor microcirculation during fractionated radiation therapy in patients with rectal carcinoma: preliminary results and implications for therapy. AB - PURPOSE: To measure microcirculatory changes during chemoirradiation and to correlate perfusion index (PI) values with therapy outcome. MATERIALS AND METHODS: Perfusion data in 11 patients with cT3 (clinical staging, tumor invaded the perirectal tissue) rectal carcinoma who underwent preoperative chemoirradiation were analyzed. Perfusion data were acquired by using a T1 mapping sequence with a whole-body magnetic resonance (MR) imager. After contrast medium was intravenously infused at a constant rate, concentration-and-time curves were evaluated for arterial blood and tumor. All patients underwent MR imaging before and at constant intervals during chemoirradiation. Clinical stages before therapy were compared with surgical stages after therapy. RESULTS: Spatial and temporal resolution on dynamic T1 maps were sufficient to reveal changes in contrast medium accumulation in the tumor. Comparison of PI values and radiation dose showed a significant increase in the 1st (P: =.003) and 2nd weeks (P: =.01) of treatment; values subsequently returned to pretreatment levels or showed a renewed increase. High initial PI values correlated with greater lymph node downstaging (P: =.042). CONCLUSION: Dynamic T1 mapping provides a suitable tool for monitoring tumor microcirculation during chemoirradiation and offers the potential for individual optimization of therapeutic procedures. Furthermore, these results indicate that the PI map may serve as a prognostic factor. PMID- 11058633 TI - US identification of the anal sphincter complex and levator ani muscle in neonates: infracoccygeal approach. AB - PURPOSE: To identify the anal sphincter complex and levator ani muscle at transperineal ultrasonography (US) with the infracoccygeal approach. MATERIALS AND METHODS: Infracoccygeal US was performed with a 7-MHz linear-array transducer in 40 healthy neonates. The babies were placed in the supine position, and both legs were drawn up to the chest. The buttocks were accordingly lifted up. The approach site was just inferior to the coccyx and posterior to the anus. Scanning was performed to obtain transverse images of the anorectal area. The thickness of the anal sphincter complex and the puborectalis muscle were measured. RESULTS: Infracoccygeal US revealed the internal anal sphincter (IAS), the external anal sphincter (EAS), and the puborectalis muscle in all babies. The IAS and EAS were depicted as inner and outer hypoechoic ringlike structures, respectively. A round, hyperechoic space (intersphincteral space) was present between the hypoechoic IAS and EAS. The puborectalis muscle was identified as a U-shaped hypoechoic structure. The bulbocavernosus and ischiocavernous muscles were also identified. Mean thicknesses were as follows: IAS, 1.3 mm +/- 0.3 (SD) (range, 0.8-1.9 mm); EAS, 1.6 mm +/- 0.3 (range, 1.2-2.3 mm); and puborectalis muscle, 1.1 mm +/- 0.3 (range, 0.6-1.9 mm). CONCLUSION: Infracoccygeal transperineal US is an excellent diagnostic modality for demonstrating the anal sphincter complex and levator ani muscle in neonates. PMID- 11058634 TI - Normal anal sphincter anatomy and age- and sex-related variations at high-spatial resolution endoanal MR imaging. AB - PURPOSE: To describe the various patterns of normal sphincter anatomy as seen at endoanal magnetic resonance (MR) imaging and to assess sex- and age-related variations in the dimensions of the anal sphincter to refine the diagnosis of sphincter disorders. MATERIALS AND METHODS: Endoanal MR imaging (1.5 T) was performed in 100 healthy volunteers (50 women, 50 men) evenly distributed between ages 20 and 85 years. The essential anatomic structures were evaluated, and various patterns in men and women were recorded. The thickness of the anal sphincter muscles and the length of the anal canal were measured, and age- and sex-related correlations were studied. RESULTS: Sex-related differences included a significantly shorter external sphincter in women than in men both laterally (mean, 27.1 mm +/- 5.4 vs 28.6 mm +/- 4.3; P: <.05) and anteriorly (mean, 14.0 mm +/- 3.0 vs 27.0 mm +/- 53.0; P: <.051). The superficial transverse perineal muscle is located more superiorly in women than in men. The central perineal tendon in men is a central muscular insertion point; in women, it represents an area where muscle fibers imbricate. Age-related variations included a significant decrease in the thickness of the external sphincter in men (P: <.01). Significant decrease in the thickness of the longitudinal muscle and increase in the thickness of the internal sphincter were noted in both sexes (P: <.01). CONCLUSION: High-spatial-resolution endoanal MR imaging provides excellent visualization of pelvic floor structures. Severe atrophy as it occurs in incontinent patients should be differentiated from physiologic, age-related thinning of the external sphincter and longitudinal muscle. PMID- 11058635 TI - Pancreas divisum and "santorinicele": diagnosis with dynamic MR cholangiopancreatography with secretin stimulation. AB - PURPOSE: To evaluate the usefulness of magnetic resonance (MR) cholangiopancreatography (MRCP) before and after secretin administration in diagnosing santorinicele in patients with pancreas divisum. MATERIALS AND METHODS: One hundred seven patients suspected of having pancreatic disease underwent MRCP before and after secretin administration (S-MRCP). S-MRCP images were evaluated for pancreas divisum and santorinicele and for size of the main pancreatic duct and santorinicele. The onset of duodenal filling was calculated on dynamic S-MRCP images. RESULTS: Pancreas divisum was detected in five (5%) of 107 patients at MRCP and in 10 (9%) of 107 patients at S-MRCP. Santorinicele was detected in three (21%) of 14 patients at MRCP and in an additional four (seven [50%] of 14) patients at S-MRCP in patients with pancreas divisum. Santorinicele was confirmed in six of seven patients at endoscopic retrograde cholangiopancreatography (ERCP); in one of seven patients, ERCP was unsuccessful. The duct of Santorini was significantly (P: <.05) larger in the pancreatic head in patients with pancreas divisum and santorinicele (3.6 mm) compared with those with only pancreas divisum (2.2 mm). A noteworthy reduction in size of the pancreatic duct (26%) and of the santorinicele (63%) was observed after sphincterotomy. The onset of duodenal filling was delayed significantly in patients with santorinicele (2.1 vs 1.3 minutes; P: <.05). CONCLUSION: S-MRCP helps in identifying pancreas divisum and santorinicele, which may be the cause of impeded pancreatic outflow. PMID- 11058636 TI - Overlooked gastric carcinoma: pitfalls in upper gastrointestinal radiology. AB - PURPOSE: To evaluate the reasons for misdiagnosis of gastric carcinoma at upper gastrointestinal radiography. MATERIALS AND METHODS: Upper gastrointestinal radiographs obtained within 3 years prior to diagnosis of gastric carcinoma in 336 patients were selected. Two radiologists who were initially blinded and then unblinded to the diagnosis reviewed the radiographs. Decisions were made by means of consensus. The reason for misdiagnosis was classified as perceptual error when the lesion was identified correctly at the blinded review, as possible perceptual error when the lesion was identified only at the unblinded review, and as technical error when the lesion could not be identified at either review and technical deficiencies were thought to be the cause. RESULTS: Twenty-four patients underwent upper gastrointestinal radiography within 3 years prior to diagnosis of 27 carcinomas. The reason for misdiagnosis was classified as perceptual error in 11, as possible perceptual error in four, and as technical error in five lesions. In the remaining seven lesions, the lesion could not be identified at either review, and technical deficiencies were not thought to be the cause. The most common overlooked finding was depression (10 of 15), and the most common presumed technical error was incomplete compression study (seven of 11). CONCLUSION: Careful attention should be paid to detect limited barium pooling during double-contrast studies to avoid overlooking depressions. PMID- 11058637 TI - Nondiffuse fatty change of the liver: discerning pseudotumor on MR images enhanced with ferumoxides-initial observations. AB - PURPOSE: To clarify the findings of nondiffuse fatty change of the liver on ferumoxides-enhanced magnetic resonance (MR) images. MATERIALS AND METHODS: Of 202 patients who underwent ferumoxides-enhanced MR imaging, eight who had nondiffuse fatty change of the liver at computed tomography (CT) were examined as study subjects. MR imaging findings before and 1 hour after ferumoxides administration were compared with CT findings. RESULTS: Focal fatty areas of the liver showing low attenuation on CT images were depicted as areas of relatively high intensity on the ferumoxides-enhanced T1-weighted images in all patients. On enhanced T2-weighted images, focal fatty change showed relatively high intensity in three and isointensity in one of the four patients. Focal spared areas appearing as areas of relatively high attenuation on CT images were depicted as areas of relatively low intensity on the ferumoxides-enhanced T1- and T2-weighted images in all patients. CONCLUSION: Although prior reports of hepatic MR imaging with ferumoxides indicated that there is accumulation of ferumoxides within focal fatty areas that are no longer seen after the administration of contrast medium, this study revealed that focal fatty change and focal spared areas of fatty liver may be pseudotumors because of the relatively high intensity of fatty areas of the liver. Radiologists can distinguish these conditions from hepatic tumors by using the opposed-phase gradient-echo sequence or the fat-saturation technique. PMID- 11058638 TI - Detection and mapping of intraabdominal adhesions by using functional cine MR imaging: preliminary results. AB - PURPOSE: To identify and map intraabdominal adhesions by using functional cine magnetic resonance (MR) Imaging. MATERIALS AND METHODS: Twenty-seven patients suspected of having intraabdominal adhesions were examined. Section-by-section dynamic depiction of induced visceral slide throughout the whole abdomen was achieved by using a transverse or sagittal true fast imaging with steady-state precession sequence. Location and type of diagnosed adhesions were documented by using the nine segments of the abdominal map. These criteria and intraoperative results were compared in 13 patients. RESULTS: MR images depicted a total of 42 intraabdominal adhesions; 21 (50%) were in the lower abdomen. The most common types of adhesions were located between the ventral abdominal wall and small bowel loops (n = 10 [24%]) and between adjacent small-bowel loops (n = 9 [21%]). Comparison with the intraoperative results showed a sensitivity of 87.5% and a specificity of 92.5%. MR imaging was most accurate in depicting adhesions to the abdominal wall (15 [94%] of 16) and subperitoneal space (eight [100%] of eight). The presence of adhesions between bowel loops was overestimated. CONCLUSION: Detection of visceral slide at functional cine MR imaging is easy to perform and represents a well-tolerated and accurate procedure for use in the identification of intraabdominal adhesions in patients with chronic pain and equivocal clinical findings. PMID- 11058639 TI - Focused abdominal US in patients with trauma. AB - PURPOSE: To evaluate the accuracy of focused abdominal ultrasonography (US) in detecting abdominal injuries that require in-hospital patient treatment in the setting of blunt abdominal trauma. MATERIALS AND METHODS: One thousand ninety patients with blunt abdominal trauma were assessed with focused abdominal US within 30 minutes of arrival at the hospital. Focused abdominal US results were positive if intra- or retroperitoneal fluid was detected. Patients with negative US results and no other major injuries were observed in the emergency department for 12 hours before discharge. Patients who deteriorated clinically after negative initial US underwent repeat US and/or emergency abdominopelvic computed tomography (CT). Patients with positive or indeterminate US results underwent emergency abdominopelvic CT. RESULTS: Nine hundred seventy-four (89%) patients had negative focused abdominal US results; eight of these underwent CT. Sixty-six (6%) had positive US results. Four (0.4%) had false-negative and 19 (1.7%) had false-positive US results. Twenty-seven (2.5%) had indeterminate US results; of these, five (18.5%) had positive CT results. One hundred twenty-four (11.4%) required emergency CT. After indeterminate cases were excluded, focused abdominal US had 94% sensitivity, 98% specificity, 78% positive predictive value, 100% negative predictive value, and 95% accuracy. CONCLUSION: Focused abdominal US has a high negative predictive value for major abdominal injury in patients with blunt abdominal trauma. PMID- 11058640 TI - Technique factors and image quality as functions of patient weight at abdominal CT. AB - PURPOSE: To investigate how changes in kilovolt peak and milliampere second settings, and patient weight affect transmitted x-ray energy fluence and the image contrast-to-noise ratio (CNR) at abdominal computed tomography (CT). MATERIALS AND METHODS: Cylinders of water were used as patient models, and x-ray spectra, including x-ray tube potentials of 80-140 kVp, were investigated. The mean photon energy and energy fluence transmitted through water cylinders with varying diameters and the image contrast for fat, muscle, bone, and iodine relative to water were determined. The effect of changing the x-ray tube potential on CNR also was investigated. RESULTS: At a constant kVp, increasing patient weight from 10 kg to 120 kg reduced the transmitted energy fluence by two orders of magnitude. Changing the x-ray tube potential from 80 kVp to 140 kVp increased the mean photon energy from approximately 52 keV to approximately 72 keV and thus reduced the image contrast relative to water by 12% for muscle, 21% for fat, 39% for bone, and 50% for iodine (approximate reduction values). Increasing the x-ray tube potential from 80 kVp to 140 kVp increased the CNR by a factor of 2.6 for muscle and by a factor of 1. 4 for iodine. CONCLUSION: With changes in patient weight at abdominal CT, x-ray tube potentials must be varied to maintain a constant detector energy fluence. Increasing the x-ray tube potential generally improves CNR. PMID- 11058641 TI - Case 32 PMID- 11058642 TI - Case 28: Proximal interruption of the right pulmonary artery. PMID- 11058643 TI - Bronchogenic cyst: imaging features with clinical and histopathologic correlation. AB - PURPOSE: To characterize the imaging features of bronchogenic cysts. MATERIALS AND METHODS: The computed tomographic (CT) and/or magnetic resonance (MR) or ultrasonographic images in 68 histopathologically proved cases of bronchogenic cyst in 38 male and 30 female patients, aged newborn to 72 years (mean, 22 years), were retrospectively reviewed. RESULTS: There were 58 mediastinal and 10 extramediastinal cysts. At CT (n = 62), 60 cysts were sharply marginated with smooth (n = 35) or lobulated (n = 25) borders. Twenty-five cysts were of water attenuation, 25 were of soft-tissue attenuation, two were air filled, two had an air-fluid level, and two had dependent milk of calcium. On T1-weighted MR images (n = 23), 18 cysts were hyperintense and five were isointense to cerebrospinal fluid. On T2-weighted MR images (n = 18), 17 cysts were isointense or hyperintense to cerebrospinal fluid. Of the 25 soft-tissue-attenuation lesions at CT, 11 appeared cystic because of internal homogeneity, lack of internal enhancement, mural enhancement, and characteristic location. Fourteen appeared solid based on morphology and attenuation. MR imaging of nine of the latter showed marked hyperintensity on T2-weighted images. CONCLUSION: CT of bronchogenic cysts typically shows sharply marginated mediastinal masses of soft tissue or water attenuation. Most appear cystic. A minority appear solid and can be confused with other lesions; MR imaging can be useful for elucidating the cystic nature of these lesions. PMID- 11058644 TI - Pulmonary embolism detection: prospective evaluation of dual-section helical CT versus selective pulmonary arteriography in 157 patients. AB - PURPOSE: To evaluate the accuracy of dual-section helical computed tomography (CT) in acute pulmonary embolism (PE) diagnosis. MATERIALS AND METHODS: Of 204 consecutive patients with clinically suspected acute PE (mean age, 58 years +/- 14 [SD]), 158 were enrolled. All patients underwent dual-section helical CT (2.7 mm effective section thickness) and selective pulmonary arteriography within 12 hours of each other. Each image was analyzed independently by two observers, who determined image quality and presence of PE among arterial segments, including at the subsegmental level. The final diagnosis was made with consensus. RESULTS: Selective pulmonary arteriography was considered optimal in 147 (93%), suboptimal in 10 (6%), and inconclusive in one (0.6%) of 158 patients. Dual-section helical CT findings were considered technically optimal in 140 (89%), suboptimal in 11 (7%), and inconclusive in six (4%). Selective pulmonary arteriography demonstrated PE in 62 patients. Four (6%) of 62 patients had isolated subsegmental PE. The sensitivity of dual-section helical CT was 90%, and the specificity was 94%. The positive and negative predictive values were 90% and 94%, respectively. CONCLUSION: Dual-section helical CT is an improvement in helical CT that offers a high sensitivity and specificity for the depiction of PE, including at the subsegmental level. Dual-section helical CT can replace pulmonary arteriography for the direct demonstration of PE in a majority of patients. PMID- 11058645 TI - Measuring performance in chest radiography. AB - PURPOSE: To use a standardized set of chest radiographs to quantify interobserver differences and to provide a basis for comparing the diagnostic performance of physicians. MATERIALS AND METHODS: A standardized set of 60 chest radiographs was presented to 162 study participants. Each participant reviewed the radiographs and recorded his or her diagnostic impression by using a fixed five-point scale. These response data were used to generate receiver operating characteristic curves and to establish performance benchmarks. The variations in performance were tested for statistical significance. RESULTS: Significant interobserver variability was identified during these assessments. The composite group of board certified radiologists demonstrated performance superior to that of the radiology residents and nonradiologist physicians. CONCLUSION: By using a receiver operating characteristic approach and a standardized set of chest radiographs, observer accuracy and variability are easily quantified. This approach provides a basis for comparing the diagnostic performance of physicians. When value is measured as a diminution in uncertainty, board-certified radiologists contribute substantial value to the diagnostic imaging system. PMID- 11058646 TI - Tracking coronary calcification by using dual-section spiral CT: a 3-year follow up. AB - PURPOSE: To investigate the accuracy of dual-section spiral computed tomography (CT) in tracking the progression of coronary calcification, as measured during a 3-year follow-up. MATERIALS AND METHODS: Two hundred forty-six patients with hypertension (mean age, 66 years +/- 6 [SD]) were preselected in accordance with the International Nifedipine Study Intervention as a Goal for Hypertension Therapy protocol. Subjects had no clinical coronary arterial disease prior to the study and no cardiovascular events during follow-up. All participants underwent baseline CT (3.2-mm section thickness; reconstruction increment, 1.5 mm) and follow-up CT after 3 years. Calcification progression was defined as any increase in total calcification score (TCS) and analyzed in accordance with five baseline TCS categories: 1-9, 10-35, 36-100, 101-250, and greater than 250. RESULTS: At baseline CT, 152 patients had a TCS greater than 0, and 106 (70%) showed progression after 3 years, while 94 had a baseline TCS of 0; of these, 26 (28%) showed progression (P: <.01 between groups). The mean TCS was significantly higher in each baseline TCS category after 3 years. The percentage increase was negatively correlated with baseline TCS (P: <.01) and ranged from 466% in the lowest category to 38% in the highest. CONCLUSION: Dual-section spiral CT depicts significant change in TCS over time and is useful in tracking calcified coronary atherosclerosis. PMID- 11058647 TI - Ductal carcinoma in situ diagnosed with stereotactic core needle biopsy: can invasion be predicted? AB - PURPOSE: To determine whether mammographic or histologic features can be used to predict which cases diagnosed as ductal carcinoma in situ (DCIS) without invasion by means of stereotactic core needle biopsy (SCNB) will have invasive disease at surgery. MATERIALS AND METHODS: From July 1992 to March 1999, DCIS without invasion was diagnosed by means of SCNB in 59 patients. Seventeen (29%) were found to have invasive disease after surgery. The underestimation rate for SCNB was compared with that obtained by means of open surgical biopsy. Mammographic and histologic features of cases with and those without invasion were compared. RESULTS: All patients had calcifications on mammograms. There was no significant difference (P: =.26) between the underestimation rate for SCNB with the 11-gauge vacuum-assisted device and that for open surgical biopsy. No statistically significant differences between cases with and those without invasion were seen in patient age, mean number of core specimens, level of suspicion, size of lesion, distribution and morphology of the calcifications, presence of an associated mass or density, subtype of DCIS, nuclear grade, or presence of necrosis or desmoplasia. CONCLUSION: Mammographic and histologic features cannot be used reliably to predict cases that are underestimated with SCNB. However, SCNB with the 11-gauge vacuum-assisted device was as reliable as open surgical biopsy for diagnosing DCIS without invasion. PMID- 11058648 TI - Radio-frequency electrocautery ablation of mammary tissue in swine. AB - PURPOSE: To establish the size, configuration, and histopathologic features of acute, subacute, and chronic radio-frequency (RF) electrocautery of mammary tissue in swine. MATERIALS AND METHODS: Eighteen RF treatments were performed in the mammary tissue of three domestic swine under ultrasonographic (US) guidance. Histopathologic examination was performed immediately after (acute animal); 2 weeks after (subacute animal); and 4 weeks after (chronic animal) treatment. RESULTS: In the acute animal, lesions were firm nodules on palpation and had a distinct line of demarcation between necrotic and viable mammary tissue (mean lesion volume, 14.24 cm(3); largest volume, 29.06 cm(3)). In the subacute animal, there was diffuse coagulation necrosis with neutrophilic infiltrates at the periphery (mean lesion volume, 6.46 cm(3); largest volume, 9.47 cm(3)), and two treatment areas had a secondary bacterial infection. In the chronic animal, lesions were still palpable and firm (mean lesion volume, 11.67 cm(3); largest volume, 25.5 cm(3)), and five of six treatment sites had an area of gray to white fibrotic tissue that blended with the surrounding tissue. However, one site had a pale yellow area of central necrosis surrounded by a fibrotic area. In both the subacute and chronic animals, two and one treatment site, respectively, had minimal areas of skin necrosis. CONCLUSION: RF ablation of breast tissue is feasible in this animal model. Problems included minimal skin erythema, residual firm treatment regions at 4 weeks, slightly variable margins of coagulation necrosis, and occasional bacterial infection. PMID- 11058649 TI - Local tumor recurrence following hepatic cryoablation: radiologic-histopathologic correlation in a rabbit model. AB - PURPOSE: To use radiologic-histopathologic correlation in an animal model to distinguish normal postoperative findings from evidence of residual tumor after cryoablation of malignant hepatic tumors. MATERIALS AND METHODS: Hepatic cryoablation was performed in 12 rabbits with VX2 tumors and in two healthy rabbits. Nonenhanced and dynamic contrast material-enhanced computed tomography (CT) and magnetic resonance (MR) imaging and power and color Doppler flow ultrasonography (US) were performed 7-8 days after cryoablation. Histopathologic findings were correlated with imaging findings. RESULTS: Twenty tumors of 5-20 mm (mean, 10 mm) and seven areas of normal liver were treated with cryolesions of 11 21 mm (mean, 15 mm). All cryolesions exhibited arterial phase rim enhancement at CT and MR imaging, and 13 (57%) of 23 lesions demonstrated peripheral flow at US because of granulation tissue. There was macroscopic recurrence in 15 (75%) of 20 treated tumors; 14 (93%) appeared as peripheral nodularity with low-grade enhancement. Necrotic tissue did not enhance. Intact vessels extended up to 6 mm inside cryolesion margins and caused focal internal enhancement and Doppler flow. Areas of high signal intensity on T2-weighted MR images correlated with liquefaction necrosis, granulation tissue, and tumor. CONCLUSION: In this animal model, recurrent tumor typically appeared as focal nodules at the cryolesion periphery. Rim and central foci of enhancement, Doppler flow, and increased signal intensity on T2-weighted MR images can be normal findings after hepatic cryoablation. PMID- 11058650 TI - Coronary sinus flow measurement by means of velocity-encoded cine MR imaging: validation by using flow probes in dogs. AB - PURPOSE: To validate coronary sinus flow measurements for quantification of global left ventricular (LV) perfusion by means of velocity-encoded cine (VEC) magnetic resonance (MR) imaging and flow probes. MATERIALS AND METHODS: Measurements of coronary sinus flow were performed in seven dogs by using VEC MR imaging at baseline, single coronary arterial stenosis, dipyridamole stress, and reactive hyperemia. These measurements were compared with flow probe measurements of coronary blood flow (CBF) in the left anterior descending coronary (LAD) and circumflex (CFX) arteries (CBF(LAD+CFX)) and coronary sinus. LV blood perfusion was calculated in milliliters per minute per gram from coronary sinus flow, and LV mass was obtained by using VEC and cine MR imaging. LV mass was validated at autopsy. RESULTS: CBF(LAD+CFX) and coronary sinus flow at VEC MR imaging showed close correlation (r = 0.98, P: <.001). The difference between CBF(LAD+CFX) and MR coronary sinus flow was 3.1 mL/min +/- 8.5 (SD). LV mass at cine MR imaging was not significantly different from that at autopsy (73.2 g +/- 12.8 vs 69. 4 g +/- 12.8). At baseline, myocardial perfusion was 0.40 mL/min/g +/- 0.09 at VEC MR imaging, and CBF(LAD+CFX) was 0.44 mL/min/g +/- 0. 08 (not significant). Reactive hyperemia resulted in 2.7- and 2. 3-fold increases in coronary sinus flow at VEC MR imaging and flow probe CBF(LAD+CFX), respectively. CONCLUSION: VEC MR imaging has the potential to measure coronary sinus flow during different physiologic conditions and can serve as a noninvasive modality to quantify global LV perfusion in patients. PMID- 11058651 TI - CT angiographic measurement of the carotid artery: optimizing visualization by manipulating window and level settings and contrast material attenuation. AB - PURPOSE: To evaluate a broad range of window and level settings for various contrast material attenuation coefficients and degrees of vascular stenosis to obtain the most accurate computed tomographic (CT) angiographic measurements. MATERIALS AND METHODS: A total of 25, 480 measurements were made transversely (perpendicular to the lumen) and by means of maximum intensity projection (MIP) in a phantom with stenoses of 0%-100%, contrast material with attenuation coefficients of 150-350 HU, and 14 window and 13 level settings. Edge definition was also evaluated. RESULTS: There was an inherent relationship between the contrast material attenuation coefficient and the optimal window and level settings in the measurement of stenoses at both transverse and MIP CT angiography. This relationship between the contrast material attenuation coefficient D: and the optimal settings for window W: and level L: was represented by the following simple equations: W:/D: = [-2 x (L:/D:)] + 1.3, where -0.2 < L:/D: < 0.5, and W:/D: = [3.3 x (L:/D:)] - 1.3, where 0.5 < L:/D: < 1.0. With a vascular contrast material attenuation coefficient of 250-350 HU, the best transverse and MIP display settings for the window and level were 96 and 150 HU, respectively. CONCLUSION: The use of optimized window and level settings at CT angiography reduces measurement variability. PMID- 11058652 TI - Intravascular MR imaging-guided balloon angioplasty with an MR imaging guide wire: feasibility study in rabbits. AB - PURPOSE: To develop a technique for intravascular magnetic resonance (MR)-guided balloon angioplasty with use of an MR imaging guide wire. MATERIALS AND METHODS: An MR imaging guide wire (0.6-mm loopless antenna) that could be placed within a balloon catheter was manufactured. The guide wire was expected to function as either an MR receiver probe in real-time MR imaging or a guide wire for use with interventional devices. Laparotomy was performed in eight rabbits, and a dilatable stenosis was created at the upper abdominal aorta. Balloon angioplasty, validated at pre- and postoperative MR aortography with renal contrast enhancement was performed by using a 1.5-T MR unit with a fast spoiled gradient echo pulse sequence, short repetition and echo times, and a rate of three frames per second. RESULTS: During MR tracking, the entire length of the MR imaging guide wire was always visible as a band of high signal intensity. In all cases, the MR imaging guide wires were passed through the aortic stenoses dilated by means of balloon inflation. Before balloon angioplasty, flow in the aorta distal to the stenosis was decreased, which caused mild contrast enhancement in each kidney. After balloon angioplasty, distal flow was restored, resulting in substantial renal enhancement. CONCLUSION: The MR imaging guide wire is a potential tool for use in endovascular interventional MR imaging. PMID- 11058653 TI - Effect of chronic, low-pressure, sterile vesicoureteral reflux on renal growth and function in a porcine model: a radiologic and pathologic study. AB - PURPOSE: To study the effects of chronic, low-pressure, sterile vesicoureteral reflux (VUR) on renal growth and function in a porcine model. MATERIALS AND METHODS: Unilateral VUR was created in five pigs, with the contralateral kidney serving as a control. Preoperatively, and 1 year later, ultrasonography, technetium 99m-dimercaptosuccinic acid (DMSA) scintigraphy, contrast material enhanced computed tomography (CT), and contrast-enhanced magnetic resonance (MR) imaging were performed. Morphologic abnormalities and relative uptake of (99m)Tc DMSA were recorded. The postcontrast enhancement ratios for parenchymal regions of interest at CT and MR imaging were determined. Ruthenium 103-labeled microspheres were used to determine regional blood flow. After the pigs were sacrificed, the kidneys were excised, weighed, and analyzed pathologically. RESULTS: Two of five refluxing kidneys had less than 45% function at scintigraphy. One of these two kidneys was small at postmortem examination. There were no other imaging or gross pathologic abnormalities. There was no significant difference in regional blood flow between the refluxing and nonrefluxing kidneys. In all of the operated on kidneys, histologic examination showed focal chronic inflammation and fibrosis. CONCLUSION: Low-pressure sterile reflux into previously normal kidneys led to mild, focal, chronic interstitial inflammation and fibrosis after 1 year. Imaging findings were normal apart from a subtle decrease in tubular function in two refluxing kidneys. PMID- 11058654 TI - Measurement of endometrial thickness at US in multicenter drug trials: value of central quality assurance reading. AB - PURPOSE: To assess the value of central quality assurance (QA) reading of transvaginal ultrasonographic (US) images obtained to measure endometrial thickness and to assess image quality. MATERIALS AND METHODS: Results of 2,000 US examinations performed in 1,000 subjects during one of two multicenter drug trials were evaluated. Endometrial thickness was measured at the study site; images were then sent to the QA center, where an experienced sonologist evaluated endometrial thickness and image quality. RESULTS: In 360 (18%) of the 2,000 examinations, image quality was insufficient for central QA reading. Repeat examinations were requested, and suggestions for improvement in technique were provided. In 349 (97%) of the 360 examinations, repeat US images were of acceptable quality. In 99 (5%) of the 1,989 examinations in which endometrial thickness was measured, central measurement of thickness differed by more than 2 mm from that of the site. In a group (n = 300) that was followed up for 1 year, requests for repeat US examinations decreased from 24% at baseline to 11% at 1 year. CONCLUSION: Central QA reading provides a consistent evaluation of endometrial thickness on US images obtained in multicenter drug trials and helps to ensure the acquisition of high-quality transvaginal US images. It further leads to demonstrable improvement in site performance. PMID- 11058655 TI - Fallopian tube disease: the cobblestone pattern as a radiographic sign. AB - PURPOSE: To identify radiographic signs of mucosal damage by comparing hysterosalpingography with salpingoscopy in a prospective study. MATERIALS AND METHODS: Forty-one candidates for laparoscopy underwent hysterosalpingography and peroperative salpingoscopy; at both, tubal patency was noted. Radiographic criteria for mucosal abnormality were rounded filling defects (ie, the cobblestone pattern) and the absence of longitudinal radiolucent bands in the ampullary tract. At salpingoscopy, tubal mucosa was categorized by means of inspection into five classes of fold pattern: classes I and II, normal; classes III-V, abnormal. Hysterosalpingographic and salpingoscopic results were compared by means of a two-by-two table and kappa statistics. RESULTS: Seventy-four tubes were evaluated. At hysterosalpingography, 31 tubes were distally nonpatent. Of these, 26 showed a distal obstruction at salpingoscopy. None of the patent tubes at hysterosalpingography showed obstruction at salpingoscopy. The agreement between hysterosalpingography and salpingoscopy in detecting abnormal mucosal pattern was 89.2% (kappa, 0.73; P: <.001). The cobblestone pattern always corresponded to intraluminal adhesions at salpingoscopy. The absence of radiolucent bands corresponded to abnormal mucosa at salpingoscopy in four of six cases. The cobblestone pattern was found only in hydrosalpinges and never in patent tubes. Six normal patent tubes at hysterosalpingography showed intraluminal adhesions at salpingoscopy. CONCLUSION: Results indicate that the cobblestone pattern is an effective radiographic sign of intraluminal adhesions in hydrosalpinges and suggest that intraluminal disease in patent tubes might not always be excluded on normal hysterosalpingograms. PMID- 11058656 TI - Cross-sectional study of osteopenia with quantitative MR imaging and bone densitometry. AB - PURPOSE: To evaluation the cancellous bone-induced intravoxel spin dephasing rate (R2') and its relationship to bone mineral density and marrow fat and to examine these parameters as predictors of vertebral fracture status. MATERIALS AND METHODS: R2' and R2, the rate constants for reversible and irreversible spin dephasing, and marrow fat fraction were measured in the lumbar vertebrae and proximal femur. One hundred thirty-nine subjects (mean age, 62.4 years +/- 11.4 [SD]; 33 men, 106 women) had spinal dual-energy x-ray absorptiometric bone mineral density (BMD) T scores ranging from +3 to -5. R2', BMD, and bone marrow composition as determinants of vertebral fracture status were examined. RESULTS: Strongest single predictors of fracture status for BMD and R2' were the Ward triangle (r(2) = 0.48) and trochanter (r(2) = 0.37), respectively. Combined, the two parameters and sites increased fracture prediction (r(2) = 0. 62), whereas the combination of multiple BMD sites did not. Multivariate regression involving marrow fat fraction further improved fracture status prediction. R2' was correlated with BMD at all sites, although slopes differed by a factor of up to 2.5, which reflected differences in trabecular orientation relative to the static field. R2, the true transverse relaxation rate, was negatively correlated with marrow fat fraction. A non-age-related increase in marrow fat fraction in osteoporosis parallels earlier findings in animal models. CONCLUSION: Cancellous bone marrow R2' measured in the proximal femur provides information, which, with BMD, improves prediction of vertebral fracture status. PMID- 11058657 TI - Shoulder appearances at MR imaging in long-term dialysis recipients. AB - PURPOSE: To examine shoulder appearances at magnetic resonance (MR) imaging in long-term dialysis recipients. MATERIALS AND METHODS: Twenty-two chronic dialysis recipients underwent 1.0-T MR imaging with a combination of T1-, T2-, and T2* weighted sequences. Rotator cuff tendon thickening was graded as present or absent by a musculoskeletal radiologist, who also measured the supraspinatus and subscapularis tendon thicknesses with electronic calipers. The long-axis dimension and location of focal osseous lesions, in addition to their T1, T2, and T2* signal intensities, were noted. RESULTS: Supraspinatus (n = 9) and subscapularis (n = 10) tendon thickening was frequently observed. Six (27%) of the 22 patients had combined thickening of the supraspinatus and subscapularis tendons without substantial involvement of the infraspinatus or teres minor tendons. These patients had undergone dialysis longer (median, 19.2 years; range, 16.3-22.8 years) than had the other patients (median, 11.7 years; range, 5.8-19.3 years; P: =.004). The 29 intraosseous lesions had high, intermediate, and low T2 signal intensity in six (21%), nine (31%), and 14 (48%) instances, respectively. CONCLUSION: Supraspinatus and/or subscapularis tendon thickening is common in chronic dialysis recipients. Bone lesions in such patients are of variable T2 signal intensity and usually subchondral or adjacent to the greater tuberosity. PMID- 11058658 TI - Treatment of Stanford type B aortic dissection with stent-grafts: preliminary results. AB - PURPOSE: To evaluate the feasibility and safety of endovascular stent-graft placement in treating Stanford type B aortic dissection. MATERIALS AND METHODS: Seven patients underwent endovascular stent-graft placement for type B aortic dissection. Five patients had acute and two had chronic dissection. In five patients, the proximal entry tear was within 2 cm of the origin of the left subclavian artery, and in two patients it was beyond this site. In three patients, the noncovered proximal portion of the stent-graft was placed across the origin of the left subclavian artery. The efficacy of the procedure was assessed at follow-up studies 3, 6, 12, and 24 months after intervention. RESULTS: The procedure was technically and clinically successful in six patients (86%). The left subclavian artery remained patent in all patients. In two patients with involvement of aortic branches, endovascular stent-graft placement restored adequate blood flow to the compromised branches. One patient was readmitted 1 month later because the dissection extended into the ascending aorta. In all but this patient, closure of the entry tear and thrombosis of the false lumen along the stent-graft were achieved. All false lumina shrank considerably. The mean follow-up time was 14 months (range, 1-25 months). CONCLUSION: Type B aortic dissections within and beyond 2 cm of the origin of the left subclavian artery can be treated safely and effectively by means of endovascular stent-graft placement. PMID- 11058659 TI - Covered retrievable expandable nitinol stents in patients with benign esophageal strictures: initial experience. AB - PURPOSE: To investigate the safety and clinical effectiveness of covered retrievable expandable nitinol stents in 25 patients with a benign esophageal stricture. MATERIALS AND METHODS: Under fluoroscopic guidance, covered retrievable expandable nitinol stents were placed in 25 patients with a benign esophageal stricture and were removed with a retrieval hook 1-8 weeks later. RESULTS: Stent placement was successful in all patients, with no procedural complications. After stent placement, all patients could ingest solid food. The stents were successfully removed from all but two patients. One patient passed the stent via the rectum, and the other regurgitated a high cervical stent. After stent removal, one patient developed a small esophagobronchial fistula, which spontaneously sealed within 1 week of stent removal. After stent removal or migration, all patients could ingest solid food. During follow-up (mean, 13 months; range, 2-25 months) after stent removal or migration, 12 patients maintained their improvement in dysphagia and needed no further treatment. Thirteen patients with recurrence were treated by means of repeat balloon dilation. CONCLUSION: Use of retrievable expandable nitinol stents seems to be a safe and effective method of treatment in selected patients with benign esophageal strictures. PMID- 11058660 TI - Hepatocellular carcinoma treated with percutaneous radio-frequency ablation: usefulness of power Doppler US with a microbubble contrast agent in evaluating therapeutic response-preliminary results. AB - PURPOSE: To evaluate the usefulness of power Doppler ultrasonography (US) with a microbubble contrast agent in assessing the therapeutic response of hepatocellular carcinomas (HCCs) treated with percutaneous radio-frequency (RF) ablation. MATERIALS AND METHODS: Forty patients with 45 nodular HCC lesions 1.0 3.8 cm in diameter underwent power Doppler US before and after intravenous injection of a microbubble contrast agent. The same procedures were repeated after US-guided percutaneous RF ablation. The results of these studies were compared with those of three-phase helical computed tomography (CT) performed immediately after RF ablation. RESULTS: Before RF ablation, nonenhanced power Doppler US demonstrated flow signals within tumor in 33 of 45 HCCs. After contrast agent administration, flow signals increased or newly appeared in all cases. After RF ablation, none of the ablated tumors showed intratumoral flow signals at nonenhanced power Doppler US, whereas six showed marginal intratumoral flow signals at contrast agent-enhanced power Doppler US. These six tumors were found to have small enhancing foci, suggestive of viable tumor, in corresponding areas at immediate follow-up CT. Additional RF ablation or transcatheter arterial chemoembolization was performed in these tumors. CONCLUSION: The results of power Doppler US with a microbubble contrast agent in HCCs treated with RF ablation correlated well with those of contrast-enhanced CT. Preliminary data suggest that contrast-enhanced power Doppler US can be a promising noninvasive technique for assessing therapeutic response. PMID- 11058661 TI - Cardiac imaging by means of electrocardiographically gated multisection spiral CT: initial experience. AB - The authors introduce a method for cardiac investigations by using electrocardiographically gated spiral scanning with a four-section computed tomographic system. Three-dimensional images were reconstructed by means of a 250 msec temporal resolution and continuous volume coverage by using a dedicated multisection cardiac volume reconstruction algorithm. Motion-free thin-section volume images were acquired with thin sections and overlapping image increments within a single breath hold. Data segment shifts in time allowed for multiphase imaging. PMID- 11058662 TI - Left ventricular function: correlation of quantitative gated SPECT and MR imaging over a wide range of values. AB - In 21 patients, the authors compared results with quantitative gated single photon emission computed tomography (SPECT) to results with magnetic resonance imaging in the assessment of left ventricular (LV) end-diastolic volume (LVEDV), end-systolic volume (LVESV), and ejection fraction (LVEF). Between the two methods, correlations were good for LVEF (r = 0.85), LVEDV (r = 0.94), and LVESV (r = 0.95). Quantitative gated SPECT can help determine LVEF, LVEDV, and LVESV. PMID- 11058663 TI - Breast biphasic compression versus standard monophasic compression in X-ray mammography. AB - Breast biphasic compression (22.5 degrees angled paddle, followed by progressive angle reduction) was compared with standard monophasic compression in x-ray mammography. The presence of the pectoral muscle was recorded for the craniocaudal (CC) view and the presence of the inframammary fold for the mediolateral oblique (MLO) view. The amount of breast in each study and image quality were assessed for both views. For all parameters, biphasic compression performed better than monophasic compression in both CC (P: <.006) and MLO (P: <.04) views. PMID- 11058664 TI - Subclavian MR arteriography: reduction of susceptibility artifact with short echo time and dilute gadopentetate dimeglumine. AB - At arterial phase gadolinium-enhanced magnetic resonance (MR) angiography, artifactual stenosis of the subclavian artery is sometimes seen adjacent to the subclavian vein on the side of the contrast material injection. Experiments in phantoms and in 19 patients showed increased artifact with longer echo time and higher concentration of injected contrast material. An effective method to substantially decrease this susceptibility artifact was threefold dilution of gadopentetate dimeglumine and use of a short echo time (1 msec). PMID- 11058665 TI - Markedly eccentric peripheral vascular stenoses: percutaneous atherectomy with an endomyocardial biopsy device. AB - In 23 patients, percutaneous atherectomy was performed with an endomyocardial biopsy device in 27 eccentric stenoses of lower limb arteries. The mean percentage stenosis was reduced from 82% to 14% (P: <.001). Histopathologic examination revealed no media or adventitial tissue in the extracted particles. The technical success and complication rates were similar to those reported for conventional percutaneous transluminal angioplasty of short symmetric lesions. PMID- 11058666 TI - Blood vessels: depiction at phase-contrast X-ray imaging without contrast agents in the mouse and rat-feasibility study. AB - The ability of phase-contrast x-ray imaging to depict blood vessels without contrast agents was tested by observing livers of a mouse and a rat with synchrotron x rays. Livers were excised by tying arteries and veins to prevent blood from flowing out of the liver. An x-ray interferometer was used to obtain x ray phase contrast. With the technique of phase-shifting x-ray interferometry, the image mapping x-ray phase shift caused by a liver was measured. The x-ray phase shift caused by blood was substantially different from that caused by other soft tissues; consequently, trees of blood vessels were revealed on the image. Vessels with diameter smaller than 0.1 mm were recognized. This result allows new possibilities for investigation of the vascular system. PMID- 11058667 TI - The authorship dilemma: will it ever be solved? PMID- 11058668 TI - Authors need to be educated on authorship principles. PMID- 11058669 TI - Presence or absence of gas in the appendix: additional criteria to rule out or confirm acute appendicitis-evaluation with US. PMID- 11058671 TI - Abstracts of current literature PMID- 11058670 TI - Primary stent placement in hemodialysis-related central venous stenoses: the dangers of a potential "radiologic dictatorship". PMID- 11058672 TI - Inhaled zanamivir for the prevention of influenza in families. Zanamivir Family Study Group. AB - BACKGROUND: As prophylaxis against influenza in families, amantadine and rimantadine have had inconsistent effectiveness, partly because of the transmission of drug-resistant variants from treated index patients. We performed a double-blind, placebo-controlled study of inhaled zanamivir for the treatment and prevention of influenza in families. METHODS: We enrolled families (with two to five members and at least one child who was five years of age or older) before the 1998-1999 influenza season. If an influenza-like illness developed in one member, the family was randomly assigned to receive either inhaled zanamivir or placebo. The family member with the index illness was treated with either 10 mg of inhaled zanamivir (163 subjects) or placebo (158) twice a day for 5 days, and the other family members received either 10 mg of zanamivir (414 subjects) or placebo (423) once a day as prophylaxis for 10 days. The primary end point was the proportion of families in which at least one household contact had symptomatic, laboratory-confirmed influenza. RESULTS: The proportion of families with at least one initially healthy household contact in whom influenza developed was smaller in the zanamivir group than in the placebo group (4 percent vs. 19 percent, P<0.001); the difference represented a 79 percent reduction in the proportion of families with at least one affected contact. Zanamivir provided protection against both influenza A and influenza B. A neuraminidase-inhibition assay and sequencing of the neuraminidase and hemagglutinin genes revealed no zanamivir-resistant variants. Among the subjects with index cases of laboratory confirmed influenza, the median duration of symptoms was 2.5 days shorter in the zanamivir group than in the placebo group (5.0 vs. 7.5 days, P=0.01). Zanamivir was well tolerated. CONCLUSIONS: When combined with the treatment of index cases, prophylactic treatment of family members with once-daily inhaled zanamivir is well tolerated and prevents the development of influenza. In this study there was no evidence of the emergence of resistant influenza variants. PMID- 11058673 TI - A comparison of levomethadyl acetate, buprenorphine, and methadone for opioid dependence. AB - BACKGROUND: Opioid dependence is a chronic, relapsing disorder with important public health implications. METHODS: In a 17-week randomized study of 220 patients, we compared levomethadyl acetate (75 to 115 mg), buprenorphine (16 to 32 mg), and high-dose (60 to 100 mg) and low-dose (20 mg) methadone as treatments for opioid dependence. Levomethadyl acetate and buprenorphine were administered three times a week. Methadone was administered daily. Doses were individualized except in the group assigned to low-dose methadone. Patients with poor responses to treatment were switched to methadone. RESULTS: There were 55 patients in each group; 51 percent completed the trial. The mean (+/-SE) number of days that a patient remained in the study was significantly higher for those receiving levomethadyl acetate (89+/-6), buprenorphine (96+/-4), and high-dose methadone (105+/-4) than for those receiving low-dose methadone (70+/-4, P<0.001). Continued participation was also significantly more frequent among patients receiving high-dose methadone than among those receiving levomethadyl acetate (P=0.02). The percentage of patients with 12 or more consecutive opioid-negative urine specimens was 36 percent in the levomethadyl acetate group, 26 percent in the buprenorphine group, 28 percent in the high-dose methadone group, and 8 percent in the low-dose methadone group (P=0.005). At the time of their last report, patients reported on a scale of 0 to 100 that their drug problem had a mean severity of 35 with levomethadyl acetate, 34 with buprenorphine, 38 with high-dose methadone, and 53 with low-dose methadone (P=0.002). CONCLUSIONS: As compared with low-dose methadone, levomethadyl acetate, buprenorphine, and high dose methadone substantially reduce the use of illicit opioids. PMID- 11058674 TI - Lack of correlation between psychological factors and subclinical coronary artery disease. AB - BACKGROUND: The relation between psychological variables and clinically evident coronary artery disease has been studied extensively, although the potential mechanisms of such a relation remain speculative. We studied the relation between multiple psychological variables and subclinical coronary artery disease to assess the possible role of such variables in atherogenesis. METHODS: We conducted a prospective study of 630 consecutive consenting, active-duty U.S. Army personnel, 39 to 45 years of age, without known coronary artery disease. Each participant was assessed for depression, anxiety, somatization, hostility, and stress. Subclinical coronary artery disease was identified by electron-beam computed tomography. RESULTS: The mean (+/-SD) age of the subjects was 42+/-2 years; 82 percent were male, and 72 percent were white. The prevalence of coronary-artery calcification was 17.6 percent (mean calcification score, 10+/ 49). The prevalence of prior or current psychiatric disorders was 12.7 percent. There was no correlation between the coronary-calcification score and the scores measuring depression (r= -0.07, P=0.08), anxiety (r=-0.07, P=0.10), hostility (r= 0.07, P=0.10), or stress (r=-0.002, P=0.96). Somatization (the number and severity of durable physical symptoms) was inversely correlated with calcification scores (r=-0.12, P=0.003), even after we controlled for age and sex. In multivariate logistic-regression models, a somatization score greater than 4 (out of a possible 26) was independently associated with the absence of any coronary-artery calcification (odds ratio, 0.49; 95 percent confidence interval, 0.25 to 0.96). CONCLUSIONS: Our data suggest that depression, anxiety, hostility, and stress are not related to coronary-artery calcification and that somatization is associated with the absence of calcification. PMID- 11058675 TI - Obesity, hypertension, and the risk of kidney cancer in men. AB - BACKGROUND: Obesity and hypertension have been implicated as risk factors for the development of renal-cell cancer. METHODS: We examined the health records of 363,992 Swedish men who underwent at least one physical examination from 1971 to 1992 and were followed until death or the end of 1995. Men with cancer (renal cell cancer in 759 and renal-pelvis cancer in 136) were identified by cross linkage of data with the nationwide Swedish Cancer Registry. Poisson regression analysis was used to estimate relative risks, with adjustments for age, smoking status, body-mass index, and diastolic blood pressure. RESULTS: As compared with men in the lowest three eighths of the cohort for body-mass index, men in the middle three eighths had a 30 to 60 percent greater risk of renal-cell cancer, and men in the highest two eighths had nearly double the risk (P for trend, <0.001). There was also a direct association between higher blood pressures and a higher risk of renal-cell cancer (P for trend, <0.001 for diastolic pressure; P for trend, 0.007 for systolic pressure). After the first five years of follow-up had been excluded to reduce possible effects of preclinical disease, the risk of renal-cell cancer was still consistently higher in men with a higher body-mass index or higher blood pressure. At the sixth-year follow-up, the risk rose further with increasing blood pressures and decreased with decreasing blood pressures, after adjustment for base-line measurements. Men who were current or former smokers had a greater risk of both renal-cell cancer and renal-pelvis cancer than men who were not smokers. There was no relation between body-mass index or blood pressure and the risk of renal-pelvis cancer. CONCLUSIONS: Higher body-mass index and elevated blood pressure independently increase the long-term risk of renal-cell cancer in men. A reduction in blood pressure lowers the risk. PMID- 11058676 TI - Images in clinical medicine. Pulmonary granulomas in an intravenous drug user. PMID- 11058677 TI - Primary immunodeficiency diseases due to defects in lymphocytes. PMID- 11058678 TI - Nonsurgical treatment of ectopic pregnancy. PMID- 11058679 TI - Influenza in the family. PMID- 11058681 TI - Correction: Drug Therapy for Breast-Feeding Women. PMID- 11058680 TI - Treating opioid dependence--new data and new opportunities. PMID- 11058682 TI - Membrane phospholipid asymmetry and signal transduction. PMID- 11058683 TI - Characterization of the increase in [Ca(2+)](i) during hypotonic shock and the involvement of Ca(2+)-activated K(+) channels in the regulatory volume decrease in human osteoblast-like cells. AB - The calcium indicator fura-2 was used to study the effect of hypotonic solutions on the intracellular calcium concentration, [Ca(2+)](i), in a human osteoblast like cell line. Decreasing the tonicity of the extracellular solution to 50% leads to an increase in [Ca(2+)](i) from approximately 150 nm up to 1.3 microm. This increase in [Ca(2+)](i) was mainly due to an influx of extracellular Ca(2+) since removing of extracellular Ca(2+) reduced this increase to approximately 250 nm. After cell swelling most of the cells were able to regulate their volume to the initial level within 800 sec. The whole-cell recording mode of the patch clamp technique was also used to study the effect of an increase in [Ca(2+)](i) on membrane currents in these cells. An increase in [Ca(2+)](i) revealed two types of Ca(2+)-activated K(+) channels, K(Ca) channels. Current through both channel types could not be observed below voltage of +80 mV with [Ca(2+)](i) buffered to 100 nm or less. With patch-electrodes filled with solutions buffering [Ca(2+)](i) to 10 microm both channels types could be readily observed. The activation of the first type was apparently voltage-independent since current could be observed over the entire voltage range used from -160 to +100 mV. In addition, the current was also blocked by charybdotoxin (CTX). The second type of K(Ca) channels in these cells could be activated with depolarizations more positive than -40 mV from a holding potential of -80 mV. This type was blocked by CTX and paxilline. Adding paxilline to the extracellular solution inhibited regulatory volume decrease (RVD), but could not abolish RVD. We conclude that two K(Ca) channel types exist in human osteoblasts, an intermediate conductance K(Ca) channel and a MaxiK-like K(Ca) channel. MaxiK channels might get activated either directly or by an increase in [Ca(2+)](i) elicited through hypotonic solutions. In combination with the volume-regulated Cl(-) conductance in the same cells this K(+) channel seems to play a vital role in volume regulation in human osteoblasts. PMID- 11058684 TI - Calcium channel current in rat dental pulp cells. AB - Voltage-gated Ca(2+) currents in early-passage rat dental pulp cells were studied using whole-cell patch-clamp techniques. With Ba(2+) as the charge carrier, two prominent inwardly-directed currents, I(f) and I(s), were identified in these cells that could be distinguished on the basis of both kinetics and pharmacology. I(f) was activated by membrane depolarizations more positive than -30 mV, and displayed fast inactivation kinetics, while I(s) was activated by steeper depolarizations and inactivated more slowly. At peak current, time constants of inactivation for I(f) and I(s) were approximately 17 vs. approximately 631 msec. Both I(f) and I(s) could be blocked by lanthanum. By contrast, only I(s) was sensitive to either Bay-K or nifedipine, a specific agonist and antagonist, respectively, of L-type Ca(2+) channels. I(s) was also blocked by the peptide omega-Conotoxin GVIA. Taken together, results suggested that I(f) was mediated by divalent cation flow through voltage-gated T-type Ca(2+) channels, whereas I(s) was mediated by L- and N-type Ca(2+) channels in the pulp cell membrane. The expression of these prominent, voltage-gated Ca(2+) channels in a presumptive mineral-inductive phenotype suggests a functional significance vis a vis differentiation of dental pulp cells for the expression and secretion of matrix proteins, and/or formation of reparative dentin itself. PMID- 11058685 TI - Candidate inhibitor of the volume-sensitive kinase regulating K-Cl cotransport: the myosin light chain kinase inhibitor ML-7. AB - K-Cl cotransport, KCC, is activated by swelling in many cells types, and promotes volume regulation by a KCl efflux osmotically coupled to water efflux. KCC is probably activated by swelling-inhibition of a kinase, permitting dephosphorylation, and activation of the cotransporter by a phosphatase. The myosin light chain kinase (MLCK) inhibitor ML-7 inhibits transporters activated by shrinkage. In red blood cells from three mammalian species, ML-7 stimulated KCC in a volume-dependent manner. Relative stimulation was greatest in more shrunken cells. Stimulation was reduced by moderate cell swelling and abolished by further swelling. The half-maximal stimulation is at approximately 20 microm ML-7, 50-fold greater than the IC(50) for inhibition of MLCK in vitro. Stimulation of KCC by ML-7 did not require cell Ca, while MLCK does. Therefore the target of ML-7 in stimulating KCC in red cells is probably not MLCK. The evidence favors stimulation of KCC by ML-7 by inhibiting the volume-sensitive kinase. Qualitatively similar effects of ML-7 on KCC in red cells from three mammalian species suggest a general mechanism. PMID- 11058686 TI - Characteristics of the quenching of 9-aminoacridine fluorescence by liposomes made from plant lipids. AB - Several laboratories have determined the surface charge density of membranes utilizing methods based on vesicle-induced quenching of the fluorescence of 9 aminoacridine and its relief by other cations. However, the computational methods by which surface charge density were calculated have not been verified in a model system. In this study, the quenching of 9-aminoacridine fluorescence by liposomes made from varying amounts of digalactosyldiacylglyceride and phosphatidic acid and relief of quenching by salts was examined. Quenching of 9-aminoacridine fluorescence increased with increasing amounts of phosphatidic acid added, independent of the composition of the added liposomes. In certain instances, the computational methods did not yield the surface charge density of the liposomes expected from their composition. However, when the effects of background ionic strength on surface potential were considered, there was a positive correlation between expected and calculated values. Therefore, the data support the contention that changes in the fluorescence of 9-aminoacridine can be used to calculate surface charge density of membranes. PMID- 11058687 TI - Spontaneous gating of olfactory cyclic-nucleotide-gated channels. AB - In vertebrates, cilia on the olfactory receptor neurons have a high density of cyclic-nucleotide-gated (CNG) channels. During transduction of odorous stimuli, cyclic AMP is formed. cAMP gates the CNG channels and this initiates the neuronal depolarization. Here it is shown that the ciliary CNG channels also open spontaneously. In the absence of odorants and second messengers, olfactory cilia have a small basal conductance to cations. Part of this conductance is similar to the cAMP-activated conductance in its sensitivity to channel inhibitors and divalent cations. The basal conductance may help to stabilize the neuronal membrane potential while limiting the sensitivity of odorant detection. PMID- 11058689 TI - Erratum. PMID- 11058688 TI - Slow gating of gap junction channels and calmodulin. AB - Certain COOH-terminus mutants of connexin32 (Cx32) were previously shown to form channels with unusual transjuctional voltage (V(j)) sensitivity when tested heterotypically in oocytes against Cx32 wild type. Junctional conductance (G(j)) slowly increased by severalfold or decreases to nearly zero with V(j) positive or negative, respectively, at mutant side, and V(j) positive at mutant side reversed CO(2)-induced uncoupling. This suggested that the CO(2)-sensitive gate might be a V(j)-sensitive slow gate. Based on previous data for calmodulin (CaM) involvement in gap junction function, we have hypothesized that the slow gate could be a CaM like pore plugging molecule (cork gating model). This study describes a similar behavior in heterotypic channels between Cx32 and each of four new Cx32 mutants modified in cytoplasmic-loop and/or COOH-terminus residues. The mutants are: ML/NN+3R/N, 3R/N, ML/NN and ML/EE; in these mutants, N or E replace M105 and L106, and N replace R215, R219 and R220. This study also reports that inhibition of CaM expression strongly reduces V(j) and CO(2) sensitivities of two of the most effective mutants, suggesting a CaM role in slow and chemical gating. PMID- 11058690 TI - Using patient-reportable clinical history factors to predict myocardial infarction. AB - Using a derivation data set of 1253 patients, we built several logistic regression and neural network models to estimate the likelihood of myocardial infarction based upon patient-reportable clinical history factors only. The best performing logistic regression model and neural network model had C-indices of 0.8444 and 0.8503, respectively, when validated on an independent data set of 500 patients. We conclude that both logistic regression and neural network models can be built that successfully predict the probability of myocardial infarction based on patient-reportable history factors alone. These models could have important utility in applications outside of a hospital setting when objective diagnostic test information is not yet be available. PMID- 11058691 TI - Multifractal texture analysis of perfusion lung scans as a potential diagnostic tool for acute pulmonary embolism. AB - A computer-assisted diagnostic (CAD) tool was developed for the diagnosis of acute pulmonary embolism (PE) in perfusion lung scans. Forty-five scans (with angiographic proof) were included in the study. The CAD tool was composed of two modules. The first module performs multifractal texture analysis on the posterior view of the perfusion scan. The second module is a decision algorithm that merges the multifractal parameters into a diagnosis regarding the presence or absence of PE. Linear and non-linear decision models were evaluated for the diagnostic task. A consensus neural network significantly outperformed all decision models including the physicians. PMID- 11058692 TI - Model-based estimation of male urethral resistance and elasticity using pressure flow data. AB - To assess urethral resistance and changes in the urethral elasticity during voiding, a lumped parameter model of the urethra was developed. The model uses pressure and flow measurements to estimate time-dependent resistance and elasticity factor. The model includes a resistance that has a function of the cross-section and urethral elasticity. Two resistivity types are compared in the constricted flow-controlling zone of the urethra: Poiseouille resistance and the Bernoulli effect. Using real pressure-flow data sets, the model was used to estimate urethral resistance and changes in urethral elasticity during voiding. Estimation of the elasticity show that in a normal patient relaxation of the urethra is a process that continues until the end of voiding. This has important implications with regard to the present methods that are used in the clinic to assess urethral obstruction or constriction. The resistance as calculated by this model, may be a useful indicator of urethral constriction and obstruction, since it is especially independent of the bladder function. Changes in the urethral elasticity during voiding which are estimated by the model add a new diagnostic parameter to pressure-flow studies. PMID- 11058693 TI - A comparison of non-linear non-parametric models for epilepsy data. AB - EEG spike and wave (SW) activity has been described through a non-parametric stochastic model estimated by the Nadaraya-Watson (NW) method. In this paper the performance of the NW, the local linear polynomial regression and support vector machines (SVM) methods were compared. The noise-free realizations obtained by the NW and SVM methods reproduced SW better than as reported in previous works. The tuning parameters had to be estimated manually. Adding dynamical noise, only the NW method was capable of generating SW similar to training data. The standard deviation of the dynamical noise was estimated by means of the correlation dimension. PMID- 11058694 TI - Multiple modeling in the study of interaction of hemodynamics and gas exchange. AB - Circulation plays an important rule in gas exchange. Therefore, there is an interaction between circulation and gas exchange. To understand the dynamic effect of these two physiological systems, a computer simulation model of hemodynamics and gas exchange is established in this work. This model includes two physiological systems, namely the respiratory and circulatory systems. It consists of five parts: the model of gas transport, exchange and storage within the body, the multi-element nonlinear mathematical model of human circulatory system, an alveolar ventilation controller, a cardiac output controller, and a controller of breathing frequency. Model simulations provide results consistent with both dynamic and steady-state responses under hypoxia. Simulation results can reflect the interaction of hemodynamics and gas exchange. Using this model, the changes of pulmonary arterial pressure and right ventricular pressure in high altitude are studied. The optimal mode of breathing extra oxygen using nasal prongs or a facial mask is studied. This model may provide a useful tool to study reaction of hypoxia and the oxygen inhalation mode under hypoxia environments. PMID- 11058695 TI - Soluble receptors for tumor necrosis factor-alpha (TNF-R p55 and TNF-R p75) in familial combined hyperlipidemia. AB - We investigated the potential role of the 75 kD receptor for tumor necrosis factor-alpha (TNF-alpha) (TNFRSF1B, located on chromosome 1 band p36.2) as a modifier gene in familial combined hyperlipidemia (FCH), based on previous linkage and association data. Age-corrected values for the soluble (s) extracellular domain of TNF-R p75 were lower in 156 well-characterized hyperlipidemic (HL) FCH relatives than in 168 normolipidemic (NL) relatives (P<0.01). Plasma concentrations of the soluble domain of the 55 kD receptor (sTNF R p55, the other TNF-alpha receptor) did not differ between HL and NL relatives. In conditional logistic regression analysis, plasma sTNF-R p75 concentration was the only non-lipid variable that contributed significantly to prediction of affected FCH status (regression coefficient=-0.413, P=0.01). The present findings have potentially important diagnostic and therapeutic implications in FCH. PMID- 11058696 TI - High prevalence of peripheral atherosclerosis in a rapidly developing country. AB - Cardiovascular disease is rapidly increasing in developing countries experiencing epidemiological transition. We investigated the prevalence of peripheral atherosclerosis in a rapidly developing country and compared our findings with data previously reported in Western populations. A cardiovascular risk factor survey was conducted in 1067 individuals aged 25-64 randomly selected from the general population of Seychelles. High-resolution ultrasonography of the right and left carotid and femoral arteries was performed in a random subgroup of 503 subjects (245 men and 258 women). In each of the four arteries, arterial wall thickness (in plaque-free segments) and atherosclerotic plaques (i.e. focal wall thickening at least 1.0 mm thick) were measured separately. The prevalence of peripheral atherosclerosis was high in this population. For instance, at least one plaque > or =1.0 mm was found in, respectively, 34.9 and 27.5% of men and women aged 25-34 and at least one plaque > or =2.5 mm was found in, respectively, 58.2 and 36.9% of men and women aged 55-64. With reference to data found in the literature, the prevalence of carotid atherosclerosis appeared to be significantly higher in Seychelles than in Western populations. This study provides further evidence for the importance of cardiovascular disease in developing countries. Determinants should be identified and relevant prevention and control programs implemented. PMID- 11058697 TI - 7beta-hydroxycholesterol induces Ca(2+) oscillations, MAP kinase activation and apoptosis in human aortic smooth muscle cells. AB - In the present study, we characterize the early cytotoxic effects of 7beta hydroxycholesterol, a major cytotoxin in oxidized LDL, in human aortic smooth muscle cells. Within a few minutes after addition, 7beta-hydroxycholesterol induced Ca(2+) oscillations with a frequency of approximately 0.3-0.4 min(-1). A few hours later, thapsigargin-sensitive Ca(2+) pools were depleted, indicating that 7beta-hydroxycholesterol perturbs intracellular Ca(2+) homeostasis. The mitogen-activated protein kinases (MAPKs) ERK1 and ERK2 (but not JNK) were activated within 5 min after addition of 7beta-hydroxycholesterol. The side-chain hydroxylated oxysterols 25-hydroxycholesterol and 27-hydroxycholesterol were more potent in inducing apoptosis than 7beta-hydroxycholesterol and cholesterol 5alpha,6alpha-epoxide, as determined by TUNEL staining. Addition of TNFalpha (10 ng/ml) and IFNgamma (20 ng/ml) enhanced the cytotoxicity of oxysterols and potentiated apoptosis. The cytokines alone were not toxic to smooth muscle cells at these concentrations. 25-Hydroxycholesterol and 7beta-hydroxycholesterol but not cholesterol inhibited protein synthesis at 4-8 h as determined by [35S]methionine incorporation assay. Morphologically, oxysterol-induced cell death was characterized by disorganization of the ER and Golgi membranes. The Ca(2+) and ERK signals preceded the ultrastructural changes induced by 7beta hydroxycholesterol. PMID- 11058698 TI - Group-II phospholipase A(2) enhances oxidized low density lipoprotein-induced macrophage growth through enhancement of GM-CSF release. AB - Inflammatory process plays an important role in the development and progression of atherosclerotic lesions. Recently, group-II phospholipase A(2) (PLA(2)), an inflammatory mediator, was reported to exist in human atherosclerotic lesions and to enhance the development of murine atherosclerotic lesions. Oxidized low density lipoprotein (Ox-LDL) stimulates the growth of several types of macrophages in vitro. Since proliferation of macrophages occurs in atherosclerotic lesions, it is possible to assume that the Ox-LDL-induced macrophage proliferation might be involved in the progression of atherosclerosis. In this study, the role of group-II PLA(2) in the Ox-LDL-induced macrophage growth was investigated using thioglycollate-elicited mouse peritoneal macrophages. Thioglycollate-elicited macrophages significantly expressed group-II PLA(2) and released it into the culture medium. The Ox-LDL-induced thymidine incorporation into thioglycollate-elicited macrophages was three times higher than that into resident macrophages, whereas under the same conditions, granulocyte/macrophage colony-stimulating factor (GM-CSF) equally induced thymidine incorporation into both types of macrophages. Moreover, the Ox-LDL induced GM-CSF release from thioglycollate-elicited macrophages was significantly higher than that from resident macrophages. In addition, the Ox-LDL-induced thymidine incorporation into macrophages obtained from human group-II PLA(2) transgenic mice and the GM-CSF release from these cells were significantly higher than those from their negative littermates, and the Ox-LDL-induced thymidine incorporation into human group-II PLA(2) transgenic macrophages was significantly inhibited by a polyclonal anti-human group-II PLA(2) antibody. These results suggest that the expression of group-II PLA(2) in thioglycollate-elicited macrophages may play an enhancing role in the Ox-LDL-induced macrophage growth through the enhancement of the GM-CSF release. PMID- 11058699 TI - Vascular smooth muscle cells preloaded with eicosapentaenoic acid and docosahexaenoic acid fail to respond to serotonin stimulation. AB - Epidemiological, animal and clinical studies indicate that n-3 fatty acids may benefit individuals with known history of cardiovascular disease or at risk of developing it. Though there is indirect evidence to suggest that the beneficial effects of n-3 fatty acids may be because of their ability to inhibit smooth muscle cell (SMC) proliferation, there are no studies that have examined this hypothesis. In this study, the mitogenic effect of serotonin (5HT) and platelet derived growth factor (PDGF), known mitogens for vascular SMC, on aortic SMCs preloaded with eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA) is examined. 5HT and PDGF could only partially stimulate proliferation of SMC that were preloaded with EPA or DHA as compared to the control cells. gamma-Linolenic acid (LA) and oleic acid (OA) did not block the 5HT or PDGF induced 3[H]thymidine incorporation suggesting that the anti-proliferative effect was specific to n-3 fatty acids only. Further, when EPA and DHA were combined in the ratio they are present in fishoils, there was a synergistic interaction in inhibiting the proliferation of SMC. Further, SMC grown in the presence of EPA or DHA, when stimulated with 5HT, failed to show an increase in 5HT(2) receptor mRNA. One of the potential mechanism by which fish oils may prevent the development of atherosclerosis or restenosis could be inhibition of the mitogen induced SMC proliferation. Combination of EPA with DHA is likely to be more beneficial. PMID- 11058700 TI - Effect of ursodeoxycholic acid on hepatic LDL binding and uptake in dietary hypercholesterolemic hamsters. AB - Administration of ursodeoxycholic acid (UDCA) has been shown to decrease serum total and low density lipoprotein (LDL) cholesterol in hypercholesterolemic patients with primary biliary cirrhosis. Results of previous studies prompted us to postulate that the cholesterol-lowering effect of UDCA may be due, at least in part, to a direct increment in hepatic LDL receptor binding [Bouscarel et al., Biochem J, 1991;280:589; Bouscarel et al., Lipids 1995;30:607]. The aim of the present investigation was to determine the ability of UDCA to enhance hepatocellular LDL receptor recruitment, as determined by its effect in vivo on LDL uptake, and its effect in vitro on LDL binding, under conditions of moderately elevated serum cholesterol. Study groups consisted of male golden Syrian hamsters fed either a standard chow diet (control), a 0.15% cholesterol containing diet, or a 0.15% cholesterol-containing diet supplemented with either 0.1% UDCA, or 0.1% chenodeoxycholic acid (CDCA). Cholesterol feeding increased (P<0.01) total serum cholesterol by 44%, and was associated with a 10-fold accumulation of cholesteryl esters in the liver (P<0.01). In vivo, hepatic uptake of [U-(14)C]sucrose-labeled hamster LDL was increased (P<0.05) to a level of 454+/-101 microl in animals fed a cholesterol-containing diet supplemented with UDCA, compared to that either without UDCA (337+/-56 microl), or with CDCA (240+/ 49 microl). The hepatic uptake of [U-(14)C]sucrose-labeled methylated human LDL, a marker of LDL receptor-independent LDL uptake, was unaffected by bile acid feeding. In vitro, specific binding of [125I]hamster LDL to isolated hepatocytes was determined at 4 degrees C, in presence and absence of 700 micromol/l UDCA. The K(D) ranged from 25 to 31 microg/ml, and was not affected by either cholesterol feeding or UDCA. In the presence of UDCA, the B(max) was increased by 19% (P<0.05) in cells isolated from control animals and by 29% (P<0.01) in cells isolated from hamsters fed a cholesterol-supplemented diet. In conclusion, in dietary hypercholesterolemic hamsters, both chronic in-vivo and acute in-vitro treatments with UDCA resulted in restoration of hepatic LDL binding and uptake to levels observed in control hamsters. PMID- 11058701 TI - The effects of lifibrol (K12.148) on the cholesterol metabolism of cultured cells: evidence for sterol independent stimulation of the LDL receptor pathway. AB - Lifibrol (4-(4'-tert. butylphenyl)-1-(4'-carboxyphenoxy)-2-butanol) is a new hypocholesterolemic compound; it effectively lowers low density lipoprotein (LDL) cholesterol. We studied the effects of lifibrol on the cholesterol metabolism of cultured cells. In the hepatoma cell line HepG2, Lifibrol decreased the formation of sterols from [14C]-acetic acid by approximately 25%. Similar to lovastatin, lifibrol had no effect on the synthesis of sterols from [14C]-mevalonic acid. Lifibrol did not inhibit 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase. Instead, cholesterol synthesis inhibition by lifibrol was entirely accounted for by competitive inhibition of HMG-CoA synthase. Lifibrol enhanced the cellular binding, uptake, and degradation of LDL in cultured cells in a dose dependent fashion. The stimulation of LDL receptors was significantly stronger than expected from the effect of lifibrol on sterol synthesis. In parallel, lifibrol increased the amount of immunologically detectable receptor protein. Stimulation of LDL receptor mediated endocytosis was observed both in the presence and in the absence of cholesterol-containing lipoproteins. In the absence of an extracellular source of cholesterol, both lifibrol and lovastatin induced microsomal HMG-CoA reductase. Co-incubation with LDL was sufficient to suppress the lifibrol mediated increase in reductase activity, indicating that lifibrol does not affect the production of the non-sterol derivative(s) which are thought to regulate HMG-CoA reductase activity at the post-transcriptional level. Considered together, the data suggest that the hypolipidemic action of lifibrol may, at least in part, be mediated by sterol-independent stimulation of the LDL receptor pathway. A potential advantage of lifibrol is that therapeutic concentrations do not interfere with the production of mevalonate which is required not only to synthesize sterols but also as a precursor of electron transport moieties, glycoproteins and farnesylated proteins. PMID- 11058702 TI - Effect of hormone replacement therapy on age-related increase in carotid artery intima-media thickness in postmenopausal women. AB - The aim of this study was to determine the changes in carotid artery intima-media thickness as measured by B-mode ultrasound in postmenopausal women receiving hormone replacement therapy (HRT) or not. One hundred and fifty-nine healthy postmenopausal women aged 45-65 years were recruited from our menopause clinic. All the selected women were free of cardio-vascular diseases and had no cardio vascular risk factors. None of the women were receiving lipid-lowering or antihypertensive drugs. Because carotid artery intima-media thickness was shown to be strongly and positively correlated with age in women aged 55 years and older but not before, women were divided into four groups according to age (<55 vs. > or =55 years) and use of HRT (current users vs. never users). All the treated women received non-oral 17beta-estradiol with a non-androgenic progestin and had started HRT within the first year after menopause. Scanning of the right common carotid artery was performed with a B-mode ultrasound imager and thickness of the intima-media complex as well as luminal diameter of the artery were determined using an automated computerized procedure. Within each age group (i.e. <55 or > or =55 years), women had comparable demographic characteristics and only differed by HRT use. Long-term treated women had significantly lower total cholesterol levels than untreated women (P=0.005). Triglycerides, low-density lipids (LDL)-cholesterol and high-density lipids (HDL)-cholesterol levels, systolic and diastolic blood pressure were not significantly different between users and non-users. In women <55 years, no significant difference in carotid intima-media thickness was found between current users (mean 2.5+/-1.4 years) and non users. In older women, the mean values of carotid intima-media thickness were significantly smaller in current users (mean 6.9+/-3.3 years) than in never treated women: 0.50+/-0.05 versus 0.56+/-0.07 mm, P<0.0001. Carotid artery intima media thickness was significantly correlated to age in never users (r=0.5, P<0.0001) but not in women who were currently receiving HRT (r=0.2, ns). These findings suggest an apparent protective effect of long-term HRT on age-related thickening of the intima-media of the right common carotid artery. This may contribute to explain the apparent cardio-protective effect of HRT after the menopause. PMID- 11058703 TI - The effect of atorvastatin on serum lipids and lipoproteins in patients with homozyous familial hypercholesterolemia undergoing LDL-apheresis therapy. AB - The efficacy of atorvastatin, a new hydroxymethylglutaryl (HMG)-CoA reductase inhibitor, in reducing serum lipid levels, modifying lipoprotein composition, and suppressing cholesterol synthesis was evaluated in patients with homozygous familial hypercholesterolemia (homozygous FH) undergoing LDL-apheresis therapy. Atorvastatin was given in escalating doses (10, 20, and 40 mg/day) to nine patients with homozygous FH. Five of nine patients responded well to atorvastatin; four of these patients were receptor-defective and the remaining one was receptor-negative. The change in LDL-cholesterol in the receptor defective patients averaged -20.6% compared to the baseline level at the highest dose of atorvastatin. Of five receptor-negative type patients, only one showed good response to atorvastatin therapy with a LDL-cholesterol reduction of 14.9%. Although the other four receptor-negative patients did not show a change in LDL cholesterol, all of them exhibited a considerable increase in HDL-cholesterol. All patients showed reduced urinary excretion of mevalonic acid, suggesting that atorvastatin decreases LDL-cholesterol by inhibiting cholesterol biosynthesis even where LDL-receptor activity is not present. Atorvastatin also decreased serum triglycerides in both receptor-negative and defective patients, especially in the latter. As cholesterol level rebounds quickly after each apheresis procedure, a combination therapy using atorvastatin and apheresis may increase the efficacy of the apheresis treatment, improving cost-benefit effectiveness by reducing the frequency of the apheresis treatment. PMID- 11058704 TI - Effect of hypertension and risk factors on diameters of abdominal aorta and common iliac and femoral arteries in middle-aged hypertensive and control subjects: a cross-sectional systematic study with duplex ultrasound. AB - There is a general tendency towards atherosclerosis and arterial dilatation in older age, and high blood pressure also tends to increase arterial diameters. The purpose of this study was to examine the effect of hypertension and other cardiovascular risk factors on aortic, common iliac and common femoral artery diameters. The diameters of the abdominal aorta and the iliac and femoral arteries and the extent of echogenic plaques in the aorta and the iliac arteries down to groin level were evaluated with ultrasound in 1007 middle-aged (40-60 years) men (505) and women (502), 496 with arterial hypertension and 511 controls. Twenty-eight subjects were excluded because of poor visualization. Men had significantly larger diameters of the abdominal aorta (mean 21.3+/-2.8 vs. 17.8+/-1.3 mm) and the common iliac (13.4+/-2.0 vs. 12.2+/-1.2) and common femoral arteries (11.0+/-1.4 vs. 9.7+/-0.9) than women (P for all <0.001), but arterial diameter was also related to the subject's size. Atherosclerotic plaques, age and height were associated with the diameter of the abdominal aorta in men, while high body mass index (BMI) had less significance. The diameter of the aorta was larger in hypertensive men aged 56-60 than in controls of the same age. In women, height, BMI and diastolic blood pressure (DBP) were associated with the diameter of the aorta, while systolic blood pressure (SBP) had less and age no effect. Age, plaques, height, BMI, DBP and SBP were associated with the diameters of the common iliac arteries in both genders, while smoking had an inverse correlation. The results on lipid values were inconsistent and an abnormal glucose tolerance test proved nonsignificant. In conclusion, arterial size measured as a diameter related to the subject's size was larger in men. Age, arterial plaques and blood pressure increased arterial diameter significantly. However, the hypertensive disease itself had only a minimal effect. The changes were smaller in women than in men. PMID- 11058705 TI - Catechin in the Mediterranean diet: vegetable, fruit or wine? AB - The aim of this study was to determine which type of diet contributes most to plasma concentration of (+)-catechin, a naturally occurring antioxidant flavonoid. Consecutive subjects (n=180) were screened. A blood sample was collected after a fasting period and (+)-catechin measurement in plasma was performed by high-performance liquid chromatography (HPLC) method using fluorescence detection. Dietary consumption of the last evening meal was assessed by a dietary recall method. Taking fruit, vegetable and wine consumption into account, four types of diet were identified. After adjustment for confounding factors, concentration of (+)-catechin in plasma was three-fold higher in diet with fruit and vegetable but without wine (449.5 microg/l), and four-fold higher in diet with wine but without vegetable and fruit (598.5 microg/l) in comparison to diet without fruit, vegetable and wine (131.6 microg/l). When the consumption of vegetable, fruit and wine was combined, the concentration was the highest (637.1 microg/l) (P<0. 001). Vegetable, fruit and wine were the major determinants of plasma (+)-catechin concentration (P<0.001). This study demonstrates that the highest plasma concentration of (+)-catechin was observed in subjects consuming fruit, vegetable and wine, and its antioxidant and antiaggregant activity could partly explain the relative protection against coronary heart disease (CHD). PMID- 11058706 TI - Do polymorphisms of apoB, LPL or apoE affect the hypocholesterolemic response to weight loss? AB - To assess whether there is a differential hypocholesterolemic response to weight loss for subjects carrying polymorphisms of the apolipoprotein B and other genes. A before and after comparison of lipid parameters following a calorie controlled diet for an intervention period of 12 weeks. A lipid clinic based in a large teaching hospital. The difference in slope coefficients relating the percentage change in lipid parameters to the change in body weight (adjusted for age, gender and initial body mass index (BMI)), for genotype subgroups defined by polymorphisms of the 5'VNTR apoB gene, two mutations of the LPL gene and ApoE. One hundred and forty six subjects completed the intervention diet. While, on average, the intervention was successful (mean weight loss 3.9%), there was no statistically significant difference in the slope coefficients relating lipid change to weight loss for most of the genotypes tested. The slope difference for long versus short 5'VNTR alleles of the apoB gene was 0.445 (-1.307, 2.198) for apolipoprotein B and -0. 104 (-1.486, 1.278) for total cholesterol. However, subjects carrying at least one varepsilon4 allele were significantly hypo responsive to weight loss, difference in slope coefficients -1.087 (-2.09, 0.084) and -1.320 (-2.589, 0.051) for total cholesterol and apoB, respectively. Although, this study is one of the largest of its kind, it has not replicated the findings of other smaller studies. These findings do not provide support for the use of genotype-targeted dietary advice in routine practice. PMID- 11058707 TI - The effect of concentrated n-3 fatty acids versus gemfibrozil on plasma lipoproteins, low density lipoprotein heterogeneity and oxidizability in patients with hypertriglyceridemia. AB - We evaluated in a double-blind randomized trial with a double-dummy design in 28 patients with primary hypertriglyceridemia, the effect of gemfibrozil (1200 mg/day) versus Omacor (4 g/day), a drug containing the n-3 fatty acids eicosapentaenoic (EPA) and docosahexaenoic acid (DHA), on lipid and lipoprotein levels, low density lipoprotein (LDL) subfraction profile and LDL oxidizability. Both Omacor and gemfibrozil therapy resulted in a similar significant decrease in serum triglyceride (TG), very low density lipoprotein (VLDL) triglyceride and VLDL cholesterol concentrations and an increase in high density lipoprotein (HDL) and LDL cholesterol concentrations. The increase in LDL cholesterol was due to a significant increase in cholesterol content of the relatively buoyant LDL subfractions LDL1, LDL2 and LDL3, whereas the relative contribution of the dense LDL subfractions LDL4 and LDL5 to total LDL tended to decrease. So, both therapies resulted in a more buoyant LDL subfraction profile, reflected by a significant increase of the value of parameter K (+10.3% on Omacor vs. +26.5% on gemfibrozil therapy, gemfibrozil vs Omacor P>0.05). Cu(2+)-induced oxidation of LDL was measured by continuous monitoring of conjugated dienes. After 12 weeks of Omacor treatment LDL appeared more prone to oxidative modification in vitro than LDL after gemfibrozil treatment, as measured by the significantly decreased lag time, preceding the onset of the lipid peroxidation. In both groups the rate of oxidation did not change with therapy. The amount of dienes formed during oxidation increased significantly on Omacor treatment, but not on gemfibrozil treatment. Plasma thiobarbituric acid reactive substances were higher after Omacor and lower after gemfibrozil treatment, although not significantly. We conclude that both Omacor and gemfibrozil have favorable effects on lipid and lipoprotein concentrations and the LDL subfraction profile. However, Omacor increased the susceptibility of LDL to oxidation, whereas gemfibrozil did not affect the resistance of LDL to oxidative modification in vitro. The clinical relevance of these changes remains to be established in the light of other postulated favorable effects of n-3 fatty acids on the course of cardiovascular disease. PMID- 11058708 TI - Gender difference in the influence of smoking on arterial wall thickness. AB - BACKGROUND: We hypothesized that arterial wall thickening, an early atherogenic alteration, might be associated with smoking differently according to gender, considering the cardiovascular protection of female sex hormones. METHODS AND RESULTS: We measured ultrasonographically carotid and femoral intima-media thickness (IMT) in 194 men and 330 women without risk factors other than smoking. In men: (i) current smokers had greater carotid and femoral IMT (P<0.01, P<0.001) and former smokers had greater femoral IMT (P<0.01) than never smokers; (ii) in pooled never, current and former smokers carotid and femoral IMT correlated to current daily smoking (P<0.01) and lifelong smoking (P<0.001); and (iii) carotid and femoral IMT correlated to age in never smokers (P<0.001), current smokers (P<0.01, P<0.001) and former smokers (P<0.01), with greater slopes in current than in former smokers at carotid site (P<0.05) and in current than in never smokers at femoral site (P<0.05). In women: (i) IMT did not differ by smoking status; (ii) in pooled smokers and non smokers femoral IMT correlated to current daily smoking (P=0.01) and to lifelong smoking (P<0.01) with a lower slope than in men (P<0.001), while carotid IMT did not; and (iii) carotid and femoral IMT correlated to age in never smokers (P<0.001), current smokers (P<0.001, P<0.05) and former smokers (P<0.001, P<0.01) with no different slopes. CONCLUSION: Smoking-related increase in IMT existed in men but not in women, suggesting a possible protection of female gender from early structural arterial alteration of smoking. PMID- 11058709 TI - Apolipoprotein E polymorphism and carotid artery intima-media thickness in a random sample of middle-aged men. AB - Genetic polymorphism of apolipoprotein E (apoE) is an important factor in the development of coronary artery disease but the results concerning apoE genotype and carotid artery atherosclerosis remain controversial. We investigated a random sample of 189 Finnish middle aged men (mean age 54 years, range 50-59) to assess the role of apoE in the process of carotid atherosclerosis. Intima-media thickness (IMT) of the carotid artery wall was measured at three standardised segments (common carotid artery, bifurcation and internal carotid artery) by B mode ultrasonography. Overall mean IMT value was also calculated. The carriers of E3/2 (n=20) genotype had significantly lower (P<0.01) total cholesterol and LDL cholesterol concentrations than carriers of E3/3 genotype (n=109) or the E4 allele (n=60). ApoE polymorphism was associated with common carotid artery IMT (P=0.034) when adjusted for age and body-mass index (model 1). The carriers of E3/2 had on average 9% (95% CI 0.8-16%, P=0.028) lower common carotid IMT values than the carriers of E3/3. After further adjustment with LDL and HDL cholesterol, systolic blood pressure, lipoprotein (a), apolipoprotein B and pack-years of smoking (model 2) the association was not statistically significant. The overall mean IMT varied significantly with apoE genotype (P=0.03 for model 1 and P=0.07 for model 2), and it was also lowest in the carriers of E3/2 genotype. This suggests that apoE E3/2 genotype is a protective factor in the development of carotid artery atherosclerosis in randomly selected middle-aged men. The favourable effect might be mediated at least partly by the lowering effect of E3/2 genotype on serum cholesterol. PMID- 11058710 TI - Apolipoprotein E polymorphism and atherosclerosis: association of the epsilon4 allele with defects in the internal elastic lamina. AB - The defects in the internal elastic lamina (IEL) have been proposed to be important for the migration of smooth muscle cells into the intima during atherosclerosis. We investigated the association of a genetic factor- apolipoprotein E (apoE) genotype--with the number of gaps in the IEL of the artery wall in 123 consecutive autopsy cases (90 male, 33 female) aged 18-93. At autopsy, the circumference of the IEL and the number of gaps in the IEL were measured in circular samples of the coeliac; (CA), superior mesenteric (SMA) and inferior mesenteric (IMA) arteries. In the series, the number of gaps per millimetre in the IEL of CA, SMA and IMA were associated with intimal thickening (P<0.0001, P=0.01 and P=0.005, respectively). In men, apoE genotype was significantly associated with the number of gaps in the IEL of the CA and IMA (P=0.033 and P=0.041, respectively). The carriers of epsilon4/3 or epsilon4/4 genotype had higher number of gaps in CA than the carriers of epsilon3/3 genotype (2.30+/-2.63 vs. 1.38+/-1.83 gaps/mm, P=0.035) and also higher number of gaps in IMA than the carriers of epsilon3/2 (2.18+/-1.71 vs. 0.66+/-0.60 gaps/mm, P=0.041). The results suggest that the apoE varepsilon4 allele may be involved with IEL fragmentation in men. This may be mediated through higher serum cholesterol associated with the varepsilon4 allele. PMID- 11058711 TI - The methylenetetrahydrofolate reductase gene is associated with increased cardiovascular risk in Japan, but not in other populations. AB - The methylenetetrahydrofolate reductase (MTHFR) gene has been associated with increased risk for cardiovascular disease in some, but not all studies. Our data sources included a MEDLINE search of the literature published before December 1998, a bibliography review, and expert consultation. Of 23 studies initially identified, 18 (9855 persons) met the inclusion criteria. Information on sample size, study design, Hardy-Weinberg equilibrium, method of genotype determination, plasma folate and homocysteine were abstracted by two reviewers using a standardized protocol. The overall odds ratio of the MTHFR gene on cardiovascular disease was estimated using the Mantel-Haenzel method. From 12 studies with angiographically-confirmed coronary artery disease (CAD), the overall odds ratio (OR) for CAD among those with heterozygous (V/A) was 1.3 (95% CI, 1.1-1.5), while it was 1.4 (1.2-1.6) for the homozygous mutant (V/V) compared to those with homozygous normal (A/A). However, the overall odds ratio for CAD among those with the V/V genotype versus A/A genotype was not statistically significant (OR: 1.1; 95% CI: 0.9-1.3) after excluding three Japanese studies. The corresponding OR for the three Japanese studies was 2.0 (1.6-2.7). For six studies with myocardial infarction (MI), the overall OR of MI was 1.0 (0.8-1.1) for those with the V/A genotype and 0.9 (0.7-1.1) for those with the V/V genotype, respectively; none of these ORs for MI was statistically significant. The MTHFR gene is associated with increased risk for CAD in Japan, but not in other populations. PMID- 11058712 TI - Correlations of elevated levels of hexacosanoate in erythrocyte membranes with risk factors for atherosclerosis. AB - We analyzed erythrocyte membrane C26:0 from 504 volunteers by high-performance liquid chromatography. The associations between the elevated levels of erythrocyte membrane C26:0 (0.20 or greater than 0.20%) and sex, obesity (body mass index> or =26.4), smoking (> or =20 cigarettes per day), present illnesses and past diseases were examined with the chi(2) test. The correlations among age and the levels of erythrocyte membrane C26:0, plasma total cholesterol, triglycerides, LDL cholesterol and HDL cholesterol were analyzed using Spearman's correlation coefficient. Moreover, the frequencies of high levels of erythrocyte membrane C26:0 were examined in male and female subjects divided into seven age groups. The elevated levels of erythrocyte membrane C26:0 were significantly more frequent in male subjects than in females, and were closely associated with obesity, smoking, and atherosclerosis-related diseases of present illnesses. The levels of erythrocyte membrane C26:0 were highly correlated with age and the levels of plasma total cholesterol, triglycerides and LDL cholesterol, and inversely with those of HDL cholesterol. The frequency of high levels of erythrocyte membrane C26:0 in male subjects was greater than that in female subjects in all of the seven age groups. Elevated levels of erythrocyte membrane C26:0 may be closely related with atherosclerosis. PMID- 11058713 TI - Enhancement of preheparin serum lipoprotein lipase mass by bezafibrate administration. AB - To clarify the clinical implication of preheparin serum lipoprotein lipase mass (preheparin LpL mass), we studied the relationships between preheparin LpL mass and serum lipids, including midband lipoproteins, which migrate between very low density lipoproteins and low density lipoproteins on polyacrylamide gel disc electrophoresis, in hyperlipidemias. And we also studied the changes of preheparin LpL mass in hypertriglyceridemic patients during bezafibrate administration, which is known to enhance LpL activity in postheparin plasma. Preheparin LpL mass correlated positively with high-density lipoprotein cholesterol (HDL-C) (r=0.418, P<0.01) and negatively with triglyceride (TG) (r= 0.256, P<0.01), but did not correlate with total cholesterol (TC) in 64 hyperlipidemic (type IIa, IIb and IV) patients. The midband lipoproteins were observed in 80% of hypertriglyceridemic patients (32/40). Preheparin LpL mass in midband lipoprotein-positive subjects was lower significantly than that in midband-negative subjects. When bezafibrate (400 mg/day) was administrated to 40 hypertriglyceridemic patients for 4 months, TG level significantly decreased ( 49+/-7%, P<0.01), TC levels decreased (-11+/-4%, not significant), and HDL-C levels increased (+27+/-4%, P<0.01). The midband lipoproteins disappeared in 95% of patients. Preheparin LpL mass significantly increased (+25+/-6%, P<0. 0005). In nine patients who stopped bezafibrate, TG levels significantly increased (+49+/-7%, P<0.01) and HDL-C levels decreased (-27+/-4%, P<0.01). Preheparin LPL mass significantly decreased (-25+/-6%, P<0.0005). These results suggested that bezafibrate administration enhanced preheparin LpL mass. And it might be implicated that enhanced LpL production by bezafibrate could reflect an increase of preheparin LpL mass. PMID- 11058714 TI - Statins and cardiovascular diseases: the multiple effects of lipid-lowering therapy by statins. AB - Cholesterol lowering involving different therapies improves the clinical outcome of patients. To define the underlying pathomechanism, we studied whether treatment with statins was associated with changes in blood thrombogenicity, endothelial dysfunction and soluble adhesion molecule levels. Fifty hypercholesterolemic patients were treated with pravastatin (40 mg/day, n=24) or simvastatin (20 mg/day, n=26). Lipid profile and blood thrombogenicity were assessed in all patients before and after 3 months of cholesterol reducing therapy. Blood thrombogenicity was assessed as thrombus formation, perfusing non anticoagulated blood directly from the patients' vein through the Badimon perfusion chamber (shear rate 1690/s). Endothelial-dependent vasomotor response was tested by laser-Doppler flowmeter. Soluble adhesion molecule level were measured by ELISA. Total and LDL cholesterol were reduced in the two treatment groups by statin therapy. Statin therapy was associated with a significant reduction in blood thrombogenicity and endothelium-dependent vasoresponse. No differences were observed between simvastatin or pravastatin treatment. Lipid lowering by statins had no effect on plasma levels of fibrinogen, sL-selectin, sP selectin and sICAM-1 antigen. Cholesterol lowering by both statins reduced the increased blood reactivity and endothelial dysfunction present under hypercholesterolemia. The multiple effects of lipid lowering therapy by statins may explain the benefits observed in recent epidemiological trials. PMID- 11058715 TI - Diabetic state induces lipid loading and altered expression and secretion of lipoprotein lipase in human monocyte-derived macrophages. AB - Non-insulin-dependent diabetes mellitus (NIDDM) is frequently associated with macroangiopathies and coronary heart diseases. Lipoprotein lipase (LPL), an enzyme known to undergo significant functional alterations in diabetic state, is also a potential atherogenic protein. Since, to the best of our knowledge, there are no data concerning LPL secreted by macrophages of NIDDM patients we conducted a study to assess the expression and activity of LPL secreted by monocyte-derived macrophages from NIDDM patients with cardiovascular complications versus cardiovascular patients without diabetes (controls). Isolated cells from NIDDM patients, after 7 days in culture in the presence of 20% autologous serum, readily exhibit a foam cell phenotype, in contrast to the cells from controls. Macrophages were mainly loaded with triglycerides, whose cellular amount was well correlated to triglyceridemia of NIDDM subjects. Concomitantly, macrophages from NIDDM patients displayed a approximately six-fold decrease of mRNA expression and a approximately two-fold reduction of the activity of secreted LPL, as compared to control cells. These data suggest that in complicated diabetic state, macrophage loading leading to foam cell formation is accelerated, at least in part, due to a diminished expression and activity of LPL. These observations add and extend the data that may explain the occurrence of accelerated atherogenesis and of the atherosclerotic complications associated with diabetes. PMID- 11058716 TI - Possible existence of platelet activation before the onset of cerebral infarction. AB - To study the existence of platelet activation before the onset of cerebral infarction, the ultrastructural features of platelets (7-day survival) and coagulation-fibrinolytic markers (70-100-min life span) were measured 2-12 h (acute phase), 7 days (subacute phase) and 6 months (chronic phase) after onset in 18 patients with cerebral infarction. Seven patients with atherosclerosis but without cerebral infarction and eight healthy subjects were studied as controls. Ultrastructural study included folds, pseudopods, vacuoles and centralization in addition to immunochemical staining such as platelet peroxidase and fibrinogen. Furthermore, beta-thromboglobulin, platelet factor-4, thrombin antithrombin complex and alpha(2)-plasmin inhibitor plasmin complex were examined as coagulation-fibrinolytic markers. Ultrastructural study of circulating platelets demonstrated no difference between acute and chronic phases and little difference between cerebral infarction and atherosclerosis, although plasma coagulation fibrinolytic markers showed an increase in cerebral infarction at the acute phase but no difference among the chronic phase of cerebral infarction, atherosclerosis and normal healthy subjects. It is considered that shape change in circulating platelets was caused by pre-existed atherosclerosis rather than the thrombotic event itself though coagulation-fibrinolytic markers were derived from the thrombotic event. PMID- 11058717 TI - The apolipoprotein A-IV-360His polymorphism determines the dietary fat clearance in normal subjects. AB - Apolipoprotein IV (apo A-IV) has been related to fat absorption and to the activation of some of the enzymes involved in lipid metabolism. Several polymorphic sites within the gene locus for apo A-IV have been detected. Previous studies have shown that the A-IV-2 isoform produces a different plasma lipid response after the consumption of diets with different fat and cholesterol content. The present study was designed to evaluate whether the apo A-IV 360His polymorphism could explain, at least in part, the interindividual variability observed during postprandial lipemia. Fifty-one healthy male volunteers (42 homozygous for the apo A-IV 360Gln allele (Gln/Gln) and nine carriers of the A-IV 360His allele), homozygous for the apo E3 allele, were subjected to a vitamin A fat load test consisting of 1 g of fat/kg body weight and 60000 IU of vitamin A. Blood was drawn at time 0 and every hour for 11 h. Plasma cholesterol (C), triacylglycerol (TG), and C, TG, apo B-100, apo B-48, apo A-IV and retinyl palmitate (RP) were determined in lipoprotein fractions. Data of postprandial lipemia revealed that subjects with the apo A-IV 360His allele had significantly greater postprandial levels in small triacylglycerol rich lipoproteins (TRL)-C (P<0.02), small TRL-TG (P<0.01) and large TRL-TG (P<0.05) than apo A-IV 360Gln/Gln subjects. In conclusion, the modifications observed in postprandial lipoprotein metabolism in subjects with the A-IV 360His allele could be involved in the different low density lipoprotein (LDL)-C responses observed in these subjects following a diet rich in cholesterol and saturated fats. PMID- 11058718 TI - Role of the macrophage galactose lectin in the uptake of desialylated LDL. AB - Desialylated low density lipoprotein (LDL) is rapidly taken up and accumulated by both peripheral blood monocytes and cells isolated from human arterial intima consisting predominantly of smooth muscle cells. It is shown that thioglycollate (TG)-elicited mouse macrophages and mouse peritoneal macrophages stimulated with lipopolysaccharide (LPS) show increased expression of a membrane-bound, galactose specific lectin that could be responsible for this uptake. In LPS-stimulated macrophages accumulation of desialylated LDL is increased ca. 2.6-fold. Accumulation of acetylated LDL in the same cells is reduced, suggesting that the galactose-specific lectin might be responsible for the uptake of desialylated LDL. Transfection of cells with the mouse macrophage Gal/GalNAc-specific lectin (MMGL) increased their capacity to take up asialofetuin (ASF) and, to a smaller extent, desialylated LDL. The uptake of desialylated LDL was small, most likely due to the high k(d) of MMGL for biantennary oligosaccharides as found on LDL, and low concentration of LDL achieved in tissue culture experiments. The data suggest that the expression of galactose-specific lectins can be elevated under inflammatory conditions, and that these receptors could contribute to foam cell formation under conditions of high desialylated LDL concentration, as might be found in arterial intima. PMID- 11058719 TI - High plasma levels of alpha- and beta-carotene are associated with a lower risk of atherosclerosis: results from the Bruneck study. AB - BACKGROUND AND PURPOSE: A large number of studies have contributed to the hypothesis that carotenoids, vitamins A and E are protective against atherosclerosis by acting as antioxidants. The aim of this study was to assess the relationship between plasma levels of carotenoids (alpha- and beta- carotene, lutein, lycopene, zeaxanthin, beta-cryptoxanthin), vitamins A and E, and atherosclerosis in the carotid and femoral arteries. METHODS: This prospective and cross sectional study involved a randomly selected population sample of 392 men and women aged 45-65 years. Carotid and femoral artery atherosclerosis was assessed by high-resolution duplex ultrasound. RESULTS: alpha- and beta- carotene plasma levels were inversely associated with the prevalence of atherosclerosis in the carotid and femoral arteries (P=0.004) and with the 5-year incidence of atherosclerotic lesions in the carotid arteries (P=0.04). These findings were obtained after adjustment for other cardiovascular risk factors (sex, age, LDL (low density lipoproteins), ferritin, systolic blood pressure, smoking, categories of alcohol consumption, social status, C-reactive protein). Atherosclerosis risk gradually decreased with increasing plasma alpha- and beta carotene concentrations (P=0.004). No associations were found between vitamin A and E plasma levels and atherosclerosis. CONCLUSIONS: This study provides further epidemiological evidence of a protective role of high alpha- and beta- carotene in early atherogenesis. PMID- 11058720 TI - Cholesterol absorption efficiency and sterol metabolism in obesity. AB - Role of enterohepatic cholesterol metabolism in obesity-induced increase of cholesterol synthesis was studied in healthy lean (BMI <24) and overweight (BMI >31) subjects by measuring serum lipids (including plant sterols, cholestanol and cholesterol precursors), cholesterol absorption % (double-label method), sterol balance and biliary lipids. New aspects of sterol metabolism in obesity were as follows: low efficiency of cholesterol absorption, reduced ratios to cholesterol of serum and biliary plant sterols and cholestanol (5alpha-derivative of cholesterol), and a marked increase of serum and biliary cholesterol precursor sterols. Percent of cholesterol absorption was positively related to serum cholestanol and plant sterols, and negatively to cholesterol synthesis, measured by the sterol balance technique or cholesterol precursor sterols in serum or bile. Total and endogenous cholesterol fluxes into the intestine were increased, but owing to low absorption percent, mass of cholesterol absorption was within control limits in the obese subjects. Thus, per gram of their large liver tissue the entry of intestinal cholesterol may even be subnormal. Percent of cholesterol absorption was insignificantly negatively (r=-0.256) related to intestinal cholesterol pool, but significantly to biliary concentrations of cholesterol (r= 0.581), bile acids (r=-0.513) and phospholipids (r=-0.469). Thus, dilution of labeled dietary cholesterol by expanded intestinal cholesterol pool could have contributed to subnormal efficiency of cholesterol absorption, or transfer of labeled dietary cholesterol from intestinal oil phase to micellar phase may be competitively inhibited by expanded biliary secretion, resulting in reduced absorption of dietary cholesterol. These mechanisms could have contributed to changes in metabolism of non-cholesterol sterols, especially of cholestanol and plant sterols. PMID- 11058721 TI - Lipoprotein (a) is associated with endothelial function in healthy postmenopausal women. AB - BACKGROUND: Lipoprotein (a) (Lp(a)) is an independent risk factor for atherosclerotic cardiovascular disease. The atherogenic potential of Lp(a) may be by impairment of endothelial function. Objectives. We investigated the relation of Lp(a) plasma levels to endothelium dependent and independent dilatation of the brachial artery in healthy postmenopausal women. METHODS: One hundred and five healthy postmenopausal women aged 52-67 years were included in the study. Endothelial function was assessed non-invasively by measuring percent lumen diameter change in the brachial artery after reactive hyperemia and sublingual nitroglycerine spray. RESULTS: Flow mediated dilatation was inversely related to the plasma logLp(a) level. Mean change per unit logLp(a) increase:-2.83% (95% CI: -5.22--0.43). Elevated Lp(a) (>239 mg/l) (upper quartile) was associated with an impaired flow mediated vasodilatation (2.4%+/-1. 2) compared to Lp(a) < or =239 mg/l (5.2%+/-0.7). Adjustment for other cardiovascular risk factors did not change the magnitude of the association. Nitroglycerine-induced vasodilatation was not significantly lower in the high Lp(a) level group, compared to the group with normal levels of Lp(a) (< or =239 mg/l) (8.0+/-1.2 vs. 11.4%+/-0.8). CONCLUSION: Elevated lipoprotein (a) levels are associated with an impaired endothelial function in healthy postmenopausal women, independent of conventional risk factors for cardiovascular disease. Since Lp(a) may be pathogenetically important for early vascular damage, elevated Lp(a) levels might contribute to the increased cardiovascular risk seen in postmenopausal women. PMID- 11058722 TI - Increased lipid peroxidation in a patient with CK-elevation and muscle pain during statin therapy. PMID- 11058723 TI - Larval growth, juvenile size and heterozygosity in laboratory reared mussels, Mytilus edulis. AB - Studies with marine bivalve juveniles have shown a positive correlation between growth and allozyme multi-locus heterozygosity (MLH), and, in some cases, between larval growth and juvenile growth, but there has been little research on the relationship between allozyme heterozygosity and larval growth. Larvae of M. edulis from different mating systems (half-sib families with a single female, or a single male parent, a reciprocal cross of two malesxtwo females and two mass matings of 13x13 and 8x17 females and males, respectively) were reared in the laboratory and selected into fast and slow growing groups when about 10-30% were undergoing metamorphosis. Offspring were reared to the juvenile stage (>3.00 mm) and both groups of each mating were electrophoresed and genotyped at up to 12 allozyme loci. There was generally good agreement with Mendelian inheritance (half-sibs and reciprocal cross) or the Hardy-Weinberg model (mass matings). Null alleles were detected at the Odh and Lap loci but there was no evidence that null allele heterozygotes grew slower than other genotypes. Over all cohorts, juveniles from the fast growing larval group were not significantly larger, or smaller, than juveniles from the slow growing group which suggests that larval growth rate may be independent of juvenile growth rate. This observation agrees with some, but not all, earlier studies and has commercial relevance. Tests of heterozygosity and juvenile shell length indicated no association between average heterozygosity across all allozyme loci and the size of juveniles in any cohort regardless of the mating system used or their larval growth rate. The association between MLH and juvenile growth in bivalves is seldom detected in cohorts from a limited genetic background. The lack of an association between heterozygosity and size might therefore be expected in the half-sib and reciprocal cross cohorts, but not in the mass matings. The results argue against any significant association between heterozygosity and larval size in mussels. PMID- 11058724 TI - Effects of reduced salinity on survival, growth, reproductive success, and energetics of the euryhaline polychaete Capitella sp. I. AB - Physiological adjustment to water of reduced salinity requires energy expenditure. In this study we sought to determine the fitness costs associated with such adjustment in the euryhaline polychaete Capitella sp. I, and the extent to which such costs could be explained by increased rates of energy expenditure. In a series of experiments conducted at 20 degrees C, salinity was reduced from 30 per thousand to either 25, 20, 15, 12, or 10 per thousand within 72 h after the larvae had been induced to metamorphose. Juveniles were reared on fine, organic-rich sediment. Over the next 15-30 days, we determined survival, growth, fecundity, and rates of respiration and feeding (via fecal pellet production). Larval salinity tolerance was also determined. Juvenile survival at salinities as low as 12-15 per thousand was comparable to that at 30 per thousand. The lower limit of salinity tolerance was 10-12 per thousand at 20 degrees C for both larvae and juveniles. Juveniles grew significantly more slowly at 12-15 per thousand in six of the seven experiments. Fecundity, however, was generally highest at intermediate salinities of 20-25 per thousand, and comparable at 30 and 15 per thousand. No individuals released embryos at 12 per thousand over the approximately 30-day observation periods in any of the three experiments in which the worms were reared at this low salinity. Reduced growth rates were not explained by differences in rates of respiration at different salinities: at reduced salinity, respiration rates were either statistically equivalent to (P>0.10) or significantly below (P<0.05) those recorded for animals maintained at 30 per thousand. Lower growth rates at lower salinities were best explained by reduced feeding rates. Further studies are required to determine whether digestive efficiency, growth hormone concentrations, or reproductive hormone concentrations are also altered by low salinity in this species. PMID- 11058725 TI - Developing the options for managing marine pests: specificity trials on the parasitic castrator, Sacculina carcini, against the European crab, Carcinus maenas, and related species. AB - The impacts of introduced marine pests are becoming increasingly apparent, prompting interest in the possibility of their biological control. We undertook laboratory and field experiments on host selection of one potential control agent (the endoparasitic barnacle, Sacculina carcini) against its natural host (the widely invasive European shore crab, Carcinus maenas) and several confamilial and more distantly related crustaceans. For comparison, we also tested host specificity in a related parasitic barnacle, Heterosaccus lunatus. The results confirm indistinct behavioral host selection in S. carcini, indicate very different mechanisms for host selection by S. carcini and H. lunatus (which could be related to differences between the two species in attachment points), and suggest host specificity in S. carcini depends on interactions between the parasite and the host's physiology. Development of convincing safety trials for marine parasites like S. carcini, in which the infective stage is a planktonic larva, will be more difficult than for many terrestrial parasites and will require detailed knowledge of the parasite's behavior and physiological interaction with its hosts. PMID- 11058726 TI - A fluctuating salinity regime mitigates the negative effects of reduced salinity on the estuarine macroalga, Enteromorpha intestinalis (L.) link. AB - We tested the response of Enteromorpha intestinalis to fluctuating reduced salinity regimes which may occur in coastal estuaries due to both natural and anthropogenic influences. In a fully crossed two factor experiment, we subjected E. intestinalis to 0, 5, 15 and 25 psu water enriched with nutrients for 1-, 5-, 11- and 23-day periods. Each period was followed by 24 h of exposure to 25 psu (ambient) water that was not nutrient enriched. Following 24 h in ambient salinity water, algae were returned to reduced salinity conditions for the appropriate period and the cycle continued over the 24 days for which all treatments were maintained. Exposure to 0 psu for 5 days or longer resulted in loss of pigmentation, decreased wet and dry biomass, increased wet wt:dry wt ratios, decreased removal of nitrogen (N) and phosphorus (P) from the water column and an accumulation of NH(4) in the water column. More frequent exposure to ambient salinity in the 1-day treatment mitigated these effects. Across all salinity levels tested, biomass increased as frequency of exposure to ambient salinity increased. At all durations of exposure to low salinity tested, biomass increased as salinity level increased. We conclude that growth of E. intestinalis is decreased by reduced salinity. E. intestinalis is able to withstand exposure to 0 psu but there is a temporal limit to this tolerance that is somewhere between 1 and 5 days. Populations of E. intestinalis in coastal estuaries may suffer from freshwater inputs if salinity conditions are persistently reduced. PMID- 11058727 TI - Ingestion and transformation of algal turf by Echinometra mathaei on Tiahura fringing reef (French Polynesia). AB - The sea urchin Echinometra mathaei is the most abundant herbivore on many tropical reefs. We studied the ingestion and digestion diel rhythms, transformation of algal turf and bioerosion attributable to this species on the Tiahura fringing reef in French Polynesia. Ingestion rates showed a circadian rhythm with most feeding taking place during the night. Absorption of food occurred throughout the day with urchins digesting food outside of the feeding period. A total of 73% of the faecal pellets consisted of CaCO(3) eroded from the reef, 20% consisted of organic matter and 7% the refractory organic matter. Of the organic matter, lipids, carbohydrates and chlorophyll were digested and absorbed and proteins were expelled in the faecal pellets. An average individual bioerosion of 0.32 g day(-1) was estimated for E. mathaei from approximately a 35 mm test diameter on the Tiahura fringing reef. We further estimated that E. mathaei release 70.5 g m(2) y(-1) of carbohydrates, 43.8 g m(2) y(-1) of lipid, 23.3 g m(2) y(-1) of protein and 2.0 g m(2) y(-1) of total chlorophyll pigments. PMID- 11058728 TI - Feeding, growth, and fecundity of Abarenicola pacifica in relation to sediment organic concentration. AB - For marine deposit-feeding invertebrates, the distribution of species with different life history strategies has long been known to be correlated with sediment organic concentration. Large populations of opportunistic species are found in sediments with enriched organic concentration, while equilibrium species populate low organic concentration sediments. Differences in both behavioral (e.g. feeding rate) and physiological (e.g. growth rate, reproductive output) adaptations determine the ability of species to establish populations in different environments. By systematically documenting differences in the way these factors vary as sediment organic concentration varies for both opportunistic and equilibrium species, we can better understand the mechanisms underlying this correlation between sediment organic concentration and species distributions. Here, we present the results of experiments examining the interactions among food concentration, feeding rate, growth rate, and reproductive output (measured as egg number and size) for the equilibrium species Abarenicola pacifica. A. pacifica is a large, long-lived, iteroparous, sub surface deposit-feeding polychaete. Individual worms were reared throughout most of one generation in sediments differing only in the concentration of organic matter. Juveniles (<20 mg AFDW) had higher feeding rates and growth rates in sediments of higher organic concentration throughout the range tested. These results are consistent with the predictions from optimal foraging theory. As worms grew, however, these patterns changed. Once worms reached a mean body size of approximately 50 mg AFDW, feeding rate was greater on sediments of lower organic concentration (although it took worms in the sediments with lower organic concentration longer to reach this size). Differences in growth rates among treatments decreased as worms grew. For worms >100 mg AFDW, growth rates were uniformly low ( approximately 1%/day) on all sediments, but the early advantage obtained by worms in the high organic treatments resulted in much greater body sizes after 200 days. Worms had higher tissue triacylglyceride concentrations and produced more eggs (independent of worm size) as sediment organic concentration increased. We conclude that A. pacifica alters its feeding rate in response to variations in food resources in such a way as to maximize its energy intake and thereby maximize fitness. Future studies should investigate whether opportunistic species (as well as other equilibrium species) also have this ability. PMID- 11058729 TI - An evaluation of lekking behavior in the fiddler crab Uca spp. AB - The reproductive behavior of Uca spp. has been extensively studied, especially the relationships between the dimorphically enlarged male claw and reproductive success. In contrast, little is known about the apparent congregations of adult males in marsh areas lacking vegetative cover where they engage in behaviors thought to attract mates. Similar congregations and displays in avian and mammalian species are termed 'leks.' In order to test the hypothesis that open area assemblages of Uca spp. are functioning as leks, we examined the sex ratios, juvenile/adult ratios, and the percentage of time that adult males spend eliciting reproductive behaviors within open and vegetated areas of marsh habitats. Moreover, to evaluate whether differences in sediment-based food resources could explain open-area aggregations, we compared substratum organic content of open and vegetated areas of marsh habitats. Substrate was also examined to determine if grain size composition varied between open and vegetated areas and might preclude the construction of breeding burrows in vegetated areas of the marsh. Three species of Uca from four marsh habitats in biogeographically distinct regions of North America were sampled including Dauphin Island, Alabama, Hunting Island, South Carolina, Saxis, Virginia, and Wallops Island, Virginia. Comparisons of male/female and juvenile/adult ratio means indicated that greater numbers of adult males occurred in open areas of all marshes. In addition, adult males allocated significantly greater time to reproductive behaviors in open rather than in vegetatively covered areas across-all biogeographic regions and among all species. Food levels (sediment organic content) in open areas were equal to or less than sediment organic contents in vegetated areas in marsh habitats at Dauphin Island and Hunting Island, the two marshes where the variable was examined. Similarly, substratum granulometry analysis revealed no significant differences between open and vegetatively covered areas of the marshes at Dauphin Island or Hunting Island that might influence choice of burrow location. Collectively, these observations support the hypothesis that lek behavior is an integral component of the reproductive repertoire in Uca spp. PMID- 11058730 TI - Impoverished second-order input to global linking in human vision. AB - Recent evidence points to the importance of global operations across spatial regions larger than individual cortical receptive fields. Studies of contour integration and motion trajectory detection suggest that network operations between local detectors underlie the encoding of extended contours in space and extended trajectories in motion. Here we ask whether such network operations also occur between second-order-detectors known to exist in visual cortex. We compared performance for stimuli composed of either first-order or second-order elements equated for visibility, and we show that unlike the first-order case, there is little or no linking interaction between local second-order detectors. Near chance performance was found for elements defined by second-order attributes when observers had to identify either an elongated spatial contour or an extended motion trajectory embedded in noise elements. This implies that the network operations thought to underlie these two global tasks receive, at best, an impoverished input from local detectors that encode second-order image attributes. PMID- 11058731 TI - Heading judgement from second-order motion. AB - We examined human heading judgement from second-order motion which was generated by random-dots with the contrast polarity determined randomly on each frame. It was found that human observers can judge heading fairly accurately from second order motion when pure translation is simulated or when self-motion toward a ground plane with gaze rotation is simulated but they cannot when self-motion toward cloud-like random dots with gaze rotations is simulated. It is suggested that the human visual system cannot decompose the flow fields into rotational and translational components by using second-order motion information alone, but it can do in some ways from the flow field of the ground plane. PMID- 11058732 TI - Opponent-color models and the influence of rod signals on the loci of unique hues. AB - To investigate how rod signals influence hue perception and how this influence can be incorporated into opponent-color models, we measured the shift of unique hue loci under dark-adapted conditions compared with cone-plateau conditions. Rod signals produced shifts of all spectral unique hues (blue, green, yellow) but in a pattern that was inconsistent with simple additive combinations of rod and cone inputs in opponent-color models. The shifts are consistent with non-linear models in which rod influence requires non-zero cone signals. Cone-signal strength may modulate or gate rod influence, or rod signals may change the gain of cone pathways. PMID- 11058733 TI - Reversed stereo depth and motion direction with anti-correlated stimuli. AB - We used anti-correlated stimuli to compare the correspondence problem in stereo and motion. Subjects performed a two-interval forced-choice disparity/motion direction discrimination task for different displacements. For anti-correlated 1d band-pass noise, we found weak reversed depth and motion. With 2d anti-correlated stimuli, stereo performance was impaired, but the perception of reversed motion was enhanced. We can explain the main features of our data in terms of channels tuned to different spatial frequencies and orientation. We suggest that a key difference between the solution of the correspondence problem by the motion and stereo systems concerns the integration of information at different orientations. PMID- 11058734 TI - Motion of contrast-modulated gratings is analysed by different mechanisms at low and at high contrasts. AB - We used a pedestal test [Lu & Sperling (1995a). Vision Research, 35, 2697-2722] to determine whether motion discrimination of contrast-modulated gratings has different properties at low contrast (4.5%) and at high contrast (45%). The amplitude-modulated gratings consisted of a 5 c/deg static carrier modulated by a moving 1 c/deg contrast envelope. We found that when contrast is low direction discrimination for contrast-modulated gratings is vulnerable to pedestals and becomes impossible at about 4 Hz. At high contrast contrast-modulated gratings are unaffected by pedestals and modulation sensitivity in a motion direction discrimination task remains high up to 12 Hz. These results are consistent with the hypothesis that separate mechanisms analyse motion of contrast-modulated gratings at low and at high contrast; at low contrast motion analysis is based on feature tracking, whereas at high contrast, contrast-modulated gratings are analysed by spatio-temporal filters. PMID- 11058735 TI - Exploring sequential stereopsis for co-planarity tasks. AB - We used the sequential stereopsis paradigm and apparatus described by Enright (Vision Research, 36, (1996) 307-312). The observer's task was to set targets to equidistance in Experiments 1-3, and to make them co-planar in Experiment 4. However, it is argued that in all experiments observers exploited a co-planarity setting strategy. Sequential stereopsis produced good performance throughout in terms of low disparity thresholds when head position was varied by rotations around three axes: vertical (azimuth condition); horizontal (elevation); and midline (tilt). It also produced good performance when the targets were shifted in position so that they both lay on one side of the median plane of the head. These results cannot be accounted for by Enright's isovergence hypothesis unless it is extended to incorporate other information about eye positions. Performance was better but not greatly so in control simultaneous stereopsis conditions, nor did it deteriorate much when the observer's view was restricted solely to the targets by removing visibility of the room in which the apparatus was located. Target settings were typically located on a concave arc centred on the median plane. This effect was quantitatively modelled using disparity correction for a relief task of co-planarity (Garding, Porrill, Mayhew, & Frisby. Vision Research, 35 (1995) 703-722). This modelling indicated over-estimations of c.10-20 cm in fixation distance for target distances in the range 71.5-112.5 cm. PMID- 11058736 TI - Evidence against a retinotopic-template matching in honeybees' pattern recognition. AB - Currently two hypotheses exist as to how insects process visual images, as photograph-like 'retinotopic-templates', or as a set of features extracted by the visual system. Several results obtained in honeybees cannot be reconciled with a retinotopic-template matching. (i) Bees discriminated between two patterns that should not be distinguished according to the template hypothesis. (ii) Bees preferred patterns that showed no overlap with the assumed template to patterns that had such an overlap. (iii) Bees showed a generalization of properties of the rewarded pattern to other patterns. Thus, in our paradigm, the bees must have used additional mechanisms and cues for the processing and classification of patterns. PMID- 11058737 TI - Suppression of displacement in severely slowed saccades. AB - Severely slowed saccades in Spinocerebellar ataxia have previously been shown to be at least partially closed-loop in nature, their long duration means that they can be modified in-flight in response to intrasaccadic target movements. In this study, a woman with these pathologically slowed saccades could modify them in flight in response to target movements, even when saccadic suppression of displacement (SSD) prevented conscious awareness of those movements. Thus, SSD is not complete, in that it provides perceptual information that is sub-threshold to consciousness but which can still be effectively utilised by the oculomotor system. PMID- 11058738 TI - Inhibition of saccade return (ISR): spatio-temporal properties of saccade programming. AB - Tasks such as reading or visual search consist of series of saccades. We have investigated to what extent saccades that are made within a series of self-paced movements are influenced by preceding movements. The present paper concerns an analysis of the duration of the fixations preceding saccades. We tested human subjects in a paradigm where they had to fixate two to four targets in a fixed order as fast as they could. We found that fixations before so-called 'return saccades' (saccades returning to the previously fixated position) are considerably longer (up to 40%) than other fixations. This phenomenon, which we call 'Inhibition of Saccade Return' (ISR), is present when return and regular saccades are mixed in one trial, and seems to be reset after each saccade. ISR is strongest at the previously fixated target, and decreases gradually from there. The radius of the area where ISR is found is about 4 degrees. The relation between ISR and 'Inhibition of Return' of spatial attention [Posner & Cohen, 1984] is discussed, as well as the neurophysiological basis of ISR. PMID- 11058739 TI - A bayesian model for the measurement of visual velocity. AB - Several models have been proposed for how the brain measures velocity from the output of motion-energy units. These models make some unrealistic assumptions such as the use of Gabor-shaped temporal filters, which are non causal, or flat spatial spectra, which are invalidated by existing data. We present a Bayesian model of velocity perception, which makes more realistic assumptions and allows the estimation of local retinal velocity regardless of the specific mathematical form of the spatial and temporal filters used. The model is consistent with several aspects of speed perception, such as the dependence of perceived speed on contrast. PMID- 11058740 TI - A transient deficit of motion perception in human. AB - We studied the motion perception abilities in a young adult, SF, who had her right occipito-temporal cortices resected to treat epilepsy. Following resection, SF showed transient deficits of both first- and second-order motion perception that recovered to normal within weeks. Previous human studies have shown either first- or second n order motion deficits that have lasted months or years after cerebral damage. SF also showed a transient defect in processing of shape-from motion with normal perception of shape from non-motion cues. Furthermore, she showed greatly increased reaction times for a mental rotation task, but not for a lexical decision task. The nature and quick recovery of the deficits in SF resembles the transient motion perception deficit observed in monkey following ibotenic acid lesions, and provides additional evidence that humans possess specialized cortical areas subserving similar motion perception functions. PMID- 11058741 TI - The fast oscillation of the electrooculogram reveals sensitivity of the human outer retina/retinal pigment epithelium to glucose level. AB - The effect of acute blood glucose elevations on human outer retinal function was examined. Electrooculograms were recorded as the background light cycled on/off with a 2-min period, eliciting rapid changes in the corneo-retinal standing potential known as the fast-oscillation of the electrooculogram. Recordings were made while subjects fasted and after they consumed 100 g of D-glucose. In all subjects, blood glucose levels strongly affected fast oscillation amplitude, which reflects photoreceptor-driven changes in RPE cell chloride concentration. The sensitivity of RPE metabolism to glucose fluctuations may relate to changes in the blood-retinal barrier that are known to occur in diabetes (e.g. macular edema). PMID- 11058742 TI - Lipocalins: unity in diversity. PMID- 11058743 TI - The lipocalin protein family: structural and sequence overview. AB - Lipocalins are remarkably diverse at the sequence level yet have highly conserved structures. Most lipocalins share three characteristic conserved sequence motifs the kernel lipocalins - while others are more divergent family members - the outlier lipocalins - typically sharing only one or two. This classification is a useful tool for analysing the family, and within these large sets are smaller groups sharing much higher levels of sequence similarity. The lipocalins are also part of a larger protein superfamily: the calycins, which includes the fatty acid binding proteins, avidins, a group of metalloproteinase inhibitors, and triabin. The superfamily is characterised by a similar structure (a repeated +1 topology beta-barrel) and by the conservation of a remarkable structural signature. PMID- 11058744 TI - Chromosomal location, exon/intron organization and evolution of lipocalin genes. AB - Lipocalins exhibit low sequence similarity that contrasts with a tightly conserved folding shared by all members of this superfamily. This conserved folding can be, at least partly, accounted for by a highly conserved gene structure. The array of lipocalin genes that have so far been studied mostly in mammals indicate a large conservation of a typical seven exon/six intron arrangement. Other conserved features include a partly coding exon 1 of variable size, fixed sizes of exons 2-5 that code for an array of lipocalin-specific beta strands and a tendency of the last exons to either fuse or expand into further exons without major changes in the length of the resulting open reading frame. The conserved exon/intron arrangement as well as a clustering of most lipocalin genes in given chromosomes of human and mouse indicate that the lipocalin genes diverged from a shared ancestor by successive rounds of duplications followed by late changes in exon arrangements. PMID- 11058745 TI - Evolution of the lipocalin family as inferred from a protein sequence phylogeny. AB - The lipocalins constitute a family of proteins that have been found in eubacteria and a variety of eukaryotic cells, where they play diverse physiological roles. It is the primary goal of this review to examine the patterns of change followed by lipocalins through their complex history, in order to stimulate scientists in the field to experimentally contrast our phylogeny-derived hypotheses. We reexamine our previous work on lipocalin phylogeny and update the phylogenetic analysis of the family. Lipocalins separate into 14 monophyletic clades, some of which are grouped in well supported superclades. The lipocalin tree was rooted with the bacterial lipocalin genes under the assumption that they have evolved from a single common ancestor with the metazoan lipocalins, and not by horizontal transfer. The topology of the rooted tree and the species distribution of lipocalins suggest that the newly arising lipocalins show a higher rate of amino acid sequence divergence, a higher rate of gene duplication, and their internal pocket has evolved towards binding smaller hydrophobic ligands with more efficiency. PMID- 11058746 TI - Experimentally determined lipocalin structures. AB - Twelve structures of distinct members of the lipocalin protein family have been solved experimentally. These structures have revolutionised our understanding of the properties of the lipocalins. Many more members of the family have been crystallised and now await structure solution. The number of solved lipocalin structures is steadily increasing, and with it increases our knowledge of this enigmatic and challenging protein family. PMID- 11058747 TI - Plasma retinol binding protein: structure and function of the prototypic lipocalin. AB - In terms of both structure and biological function, retinol binding protein (RBP) is one of the best characterized members of the lipocalin superfamily. The molecular interactions in which RBP participates are described herein. PMID- 11058748 TI - The transthyretin-retinol-binding protein complex. AB - Transthyretin (TTR, formerly called prealbumin), one of the transporters of the hormone thyroxine and the lipocalin retinol-binding protein (RBP), the specific carrier of the vitamin, are known to form, under physiological conditions, a macromolecular complex that is believed to play an important physiological role: prevention of glomerular filtration of the low molecular weight RBP in the kidneys. The physiological significance of complex formation is discussed first, followed by a brief description of the three-dimensional structure of the two participating proteins. The two X-ray models of the complex available are subsequently discussed and compared and finally the non-crystallographic evidence that supports these models is reviewed. PMID- 11058749 TI - The bacterial lipocalins. AB - The lipocalins were once regarded as a eukaryotic protein family, but new members have been recently discovered in bacteria. The first bacterial lipocalin (Blc) was identified in Escherichia coli as an outer membrane lipoprotein expressed under conditions of environmental stress. Blc is distinguished from most lipocalins by the absence of intramolecular disulfide bonds, but the presence of a membrane anchor is shared with two of its closest homologues, apolipoprotein D and lazarillo. Several common features of the membrane-anchored lipocalins suggest that each may play an important role in membrane biogenesis and repair. Additionally, Blc proteins are implicated in the dissemination of antibiotic resistance genes and in the activation of immunity. Recent genome sequencing efforts reveal the existence of at least 20 bacterial lipocalins. The lipocalins appear to have originated in Gram-negative bacteria and were probably transferred horizontally to eukaryotes from the endosymbiotic alpha-proteobacterial ancestor of the mitochondrion. The genome sequences also reveal that some bacterial lipocalins exhibit disulfide bonds and alternative modes of subcellular localization, which include targeting to the periplasmic space, the cytoplasmic membrane, and the cytosol. The relationships between bacterial lipocalin structure and function further illuminate the common biochemistry of bacterial and eukaryotic cells. PMID- 11058750 TI - Plant lipocalins: violaxanthin de-epoxidase and zeaxanthin epoxidase. AB - Violaxanthin de-epoxidase and zeaxanthin epoxidase catalyze the interconversions between the carotenoids violaxanthin, antheraxanthin and zeaxanthin in plants. These interconversions form the violaxanthin or xanthophyll cycle that protects the photosynthetic system of plants against damage by excess light. These enzymes are the first reported lipocalin proteins identified from plants and are only the second examples of lipocalin proteins with enzymatic activity. This review summarizes the discovery and characterization of these two unique lipocalin enzymes and examines the possibility of other potential plant lipocalin proteins. PMID- 11058751 TI - Tick histamine-binding proteins: lipocalins with a second binding cavity. AB - Tick histamine-binding proteins (HBPs) are lipocalins with two binding pockets. One of these binds histamine with a high affinity and is found at the position expected from other lipocalins, adjacent to the omega-loop at the open-end of the beta-barrel. A second binding cavity, which is a low-affinity site for histamine in one of the HBPs, is located at the end of the barrel that is closed off in other lipocalins. In order to create the second site, the 'closed-end' region has undergone a major reconstruction. Typical lipocalin characteristics, such as the 3(10) helix and a structural cluster of highly conserved residues, have been lost, while an alpha-helix now shields the cavity from the exterior. The prominence of acidic residues in the binding pockets is another distinctive characteristic of HBPs. Whereas most lipocalins have highly hydrophobic binding cavities designed to bind lipophilic compounds, HBPs have evolved to trap cationic, hydrophilic molecules. PMID- 11058752 TI - Lazarillo, a neuronal lipocalin in grasshoppers with a role in axon guidance. AB - In this report we present a review on the grasshopper lipocalin Lazarillo with special emphasis on how its molecular properties could account for its known function: the guidance of pioneer neurons during nervous system development. The expression and function of Lazarillo in a subset of developing neurons, its heavy glycosylation and its glycosylphosphatidylinositol linkage to the plasma membrane, make Lazarillo a unique member of the lipocalin family. We have built a model of the tertiary structure of Lazarillo in which we have studied the exposed surfaces in search for clues about ligand and protein interactions with Lazarillo. Our hypotheses about how this lipocalin can exert its function are discussed. PMID- 11058753 TI - Nitrophorins and related antihemostatic lipocalins from Rhodnius prolixus and other blood-sucking arthropods. AB - Recent gene sequence and crystal structure determinations of salivary proteins from several blood-sucking arthropods have revealed an unusual evolutionary relationship: many such proteins derive their functions from lipocalin protein folds. Many blood-sucking arthropods have independently evolved the ability to overcome a host organism's means of preventing blood loss (called hemostasis). Most blood feeders have proteins that induce vasodilation, inhibit blood coagulation, and reduce inflammation, but do so by distinctly different mechanisms. Despite this diversity, in many cases the antihemostatic activities in such organisms reside in proteins with lipocalin folds. Thirteen such lipocalins are described in this review, with a particular focus on the heme containing nitrophorins from Rhodnius prolixus, which transport nitric oxide, sequester histamine, and disrupt blood coagulation. Also described are the antiplatelet compounds RPAI, moubatin, and pallidipin from R. prolixus, Ornithodoros moubata, and Triatoma pallidipennis; the antithrombin protein triabin from T. pallidipennis; and the tick histamine binding proteins from Rhipicephalus appendiculatus. PMID- 11058754 TI - Amphibian choroid plexus lipocalin, Cpl1. AB - Choroid plexus lipocalin 1 (Cpl1) has been isolated from the African clawed toad (Xenopus laevis) and the cane toad (Bufo marinus). Xcpl1 has been used as a marker for studying early neural development. Due to its retinoid binding properties and the fact that it causes dysmorphogenesis when overexpressed in the early embryo, the protein product is considered to be part of the retinoic acid signalling pathway. Later in development and during adulthood, the epithelial cell sheet of the choroid plexus which forms the blood-cerebrospinal fluid barrier expresses cpl1 as the predominant secretory protein. These data, the similarity of Cpl1 to prostaglandin D(2) synthase and its functional homology to transthyretin will be discussed. PMID- 11058755 TI - Ex-FABP: a fatty acid binding lipocalin developmentally regulated in chicken endochondral bone formation and myogenesis. AB - Extracellular fatty acid binding protein (Ex-FABP) is a 21 kDa lipocalin specifically binding fatty acids, expressed during chicken embryo development in hypertrophic cartilage, in muscle fibers and in blood granulocytes. In chondrocyte and myoblast cultures Ex-FABP expression is increased by inflammatory agents and repressed by anti-inflammatory agents. In adult cartilage Ex-FABP is expressed only in pathological conditions such as in dyschondroplastic and osteoarthritic chickens. The possible mammalian counterpart is the Neu-related lipocalin (NRL), a lipocalin overexpressed in rat mammary cancer; NRL is homologous to the human neutrophil gelatinase associated lipocalin (NGAL) expressed in granulocytes and in epithelial cells in inflammation and malignancy and to the Sip24 (super-inducible protein 24), an acute phase lipocalin expressed in mouse after turpentine injection. Immunolocalization and in situ hybridization showed that NRL/NGAL is expressed in hypertrophic cartilage, in forming skeletal muscle fibers and in developing heart. In adult cartilage NRL/NGAL was expressed in articular cartilage from osteoarthritic patients and in chondrosarcoma. Moreover, NRL was induced in chondrocyte and myoblast cultures by an inflammatory agent. We propose that these lipocalins (Ex-FABP, NRL/NGAL, Sip24) represent stress proteins physiologically expressed in tissues where active remodeling is taking place during development and also present in tissues characterized by an acute phase response due to pathological conditions. PMID- 11058756 TI - The core lipocalin, bovine beta-lactoglobulin. AB - The lipocalin family became established shortly after the structural similarity was noted between plasma retinol binding protein and the bovine milk protein, beta-lactoglobulin. During the past 60 years, beta-lactoglobulin has been studied by essentially every biochemical technique available and so there is a huge literature upon its properties. Despite all of these studies, no specific biological function has been ascribed definitively to the protein, although several possibilities have been suggested. During the processing of milk on an industrial scale, the unpredictable nature of the process has been put down to the presence of beta-lactoglobulin and certainly the whey protein has been implicated in the initiation of aggregation that leads to the fouling of heat exchangers. This short review of the properties of the protein will concentrate mainly on studies carried out under essentially physiological conditions and will review briefly some of the possible functions for the protein that have been described. PMID- 11058757 TI - Glycodelin: a reproduction-related lipocalin. AB - Glycodelin, a human lipocalin, is a major endometrial protein with at least two differentially glycosylated isoforms. Glycodelin-A (GdA) is purified from human mid-trimester amniotic fluid, where it is secreted from the decidualized endometrium. Glycodelin-S (GdS) is synthesized in the male reproductive tract, mainly in the seminal vesicles, and secreted into seminal plasma. These two glycodelin isoforms, glycosylated in a completely different manner, serve as a good model for studying the effects of glycosylation on protein function and physicochemical properties. We have reviewed here the structure, expression and biological functions of glycodelin. PMID- 11058758 TI - Alpha-1-acid glycoprotein. AB - Alpha-1-acid glycoprotein (AGP) or orosomucoid (ORM) is a 41-43-kDa glycoprotein with a pI of 2.8-3.8. The peptide moiety is a single chain of 183 amino acids (human) or 187 amino acids (rat) with two and one disulfide bridges in humans and rats,respectively. The carbohydrate content represents 45% of the molecular weight attached in the form of five to six highly sialylated complex-type-N linked glycans. AGP is one of the major acute phase proteins in humans, rats, mice and other species. As most acute phase proteins, its serum concentration increases in response to systemic tissue injury, inflammation or infection, and these changes in serum protein concentrations have been correlated with increases in hepatic synthesis. Expression of the AGP gene is controlled by a combination of the major regulatory mediators, i.e. glucocorticoids and a cytokine network involving mainly interleukin-1 beta (IL-1 beta), tumour necrosis factor-alpha (TNF alpha), interleukin-6 and IL-6 related cytokines. It is now well established that the acute phase response may take place in extra-hepatic cell types, and may be regulated by inflammatory mediators as observed in hepatocytes. The biological function of AGP remains unknown; however,a number of activities of possible physiological significance, such as various immunomodulating effects, have been described. AGP also has the ability to bind and to carry numerous basic and neutral lipophilic drugs from endogenous (steroid hormones) and exogenous origin; one to seven binding sites have been described. AGP can also bind acidic drugs such as phenobarbital. The immunomodulatory as well as the binding activities of AGP have been shown to be mostly dependent on carbohydrate composition. Finally, the use of AGP transgenic animals enabled to address in vivo, functionality of responsive elements and tissue specificity, as well as the effects of drugs that bind to AGP and will be an useful tool to determine the physiological role of AGP. PMID- 11058759 TI - alpha(1)-Microglobulin: a yellow-brown lipocalin. AB - alpha(1)-Microglobulin, also called protein HC, is a lipocalin with immunosuppressive properties. The protein has been found in a number of vertebrate species including frogs and fish. This review summarizes the present knowledge of its structure, biosynthesis, tissue distribution and immunoregulatory properties. alpha(1)-Microglobulin has a yellow-brown color and is size and charge heterogeneous. This is caused by an array of small chromophore prosthetic groups, attached to amino acid residues at the entrance of the lipocalin pocket. A gene in the lipocalin cluster encodes alpha(1)-microglobulin together with a Kunitz-type proteinase inhibitor, bikunin. The gene is translated into the alpha(1)-microglobulin-bikunin precursor, which is subsequently cleaved and the two proteins secreted to the blood separately. alpha(1)-Microglobulin is found in blood and in connective tissue in most organs. It is most abundant at interfaces between the cells of the body and the environment, such as in lungs, intestine, kidneys and placenta. alpha(1)-Microglobulin inhibits immunological functions of white blood cells in vitro, and its distribution is consistent with an anti-inflammatory and protective role in vivo. PMID- 11058760 TI - Apolipoprotein D. AB - Apolipoprotein D (apoD) is a 29-kDa glycoprotein that is primarily associated with high density lipoproteins in human plasma. It is an atypical apolipoprotein and, based on its primary structure, apoD is predicted to be a member of the lipocalin family. Lipocalins adopt a beta-barrel tertiary structure and transport small hydrophobic ligands. Although apoD can bind cholesterol, progesterone, pregnenolone, bilirubin and arachidonic acid, it is unclear if any, or all of these, represent its physiological ligands. The apoD gene is expressed in many tissues, with high levels of expression in spleen, testes and brain. ApoD is present at high concentrations in the cyst fluid of women with gross cystic disease of the breast, a condition associated with increased risk of breast cancer. It also accumulates at sites of regenerating peripheral nerves and in the cerebrospinal fluid of patients with neurodegenerative conditions, such as Alzheimer's disease. ApoD may, therefore, participate in maintenance and repair within the central and peripheral nervous systems. While its role in metabolism has yet to be defined, apoD is likely to be a multi-ligand, multi-functional transporter. It could transport a ligand from one cell to another within an organ, scavenge a ligand within an organ for transport to the blood or could transport a ligand from the circulation to specific cells within a tissue. PMID- 11058761 TI - Human complement protein C8 gamma. AB - Human C8 gamma is a 22 kDa subunit of complement component C8, which is one of five components (C5b, C6, C7, C8, C9) that interact to form the cytolytic membrane attack complex (MAC) of complement. C8 contains three nonidentical subunits (alpha, beta, gamma) that are products of different genes. These subunits are arranged asymmetrically to form a disulfide-linked C8 alpha-gamma dimer that is noncovalently associated with C8 beta. C8 alpha and C8 beta are homologous to C6, C7 and C9 and together these proteins comprise what is referred to as the 'MAC protein family'. By comparison, C8 gamma is distinct in that it belongs to the lipocalin family of small, secreted proteins which have the common ability to bind small hydrophobic ligands. While specific roles have been identified for C8 alpha and C8 beta in the formation and function of the MAC, a function for C8 gamma and the identity of its ligand are unknown. This review summarizes the current status of C8 gamma structure and function and the progress made from efforts to determine its role in the complement system. PMID- 11058762 TI - Epididymal retinoic acid-binding protein. PMID- 11058763 TI - Major urinary proteins, alpha(2U)-globulins and aphrodisin. AB - The major urinary proteins (MUPs) are proteins secreted by the liver and filtered by the kidneys into the urine of adult male mice and rats, the MUPs of rats being also referred to as alpha(2U)-globulins. The MUP family also comprises closely related proteins excreted by exocrine glands of rodents, independently of their sex. The MUP family is an expression of a multi-gene family. There is complex hormonal and tissue-specific regulation of MUP gene expression. The multi-gene family and its outflow are characterized by a polymorphism which extends over species, strains, sexes, and individuals. There is evidence of evolutionary conservation of the genes and their outflow within the species and evidence of change between species. MUPs share the eight-stranded beta-barrel structure lining a hydrophobic pocket, common to lipocalins. There is also a high degree of structural conservation between mouse and rat MUPs. MUPs bind small natural odorant molecules in the hydrophobic pocket with medium affinity in the 10(4) 10(5) M(-1) range, and are excreted in the field, with bound odorants. The odorants are then released slowly in air giving a long lasting olfactory trace to the spot. MUPs seem to play complex roles in chemosensory signalling among rodents, functioning as odorant carriers as well as proteins that prime endocrine reactions in female conspecifics. Aphrodisin is a lipocalin, found in hamster vaginal discharge, which stimulates male copulatory behaviour. Aphrodisin does not seem to bind odorants and no polymorphism has been shown. Both MUPs and aphrodisin stimulate the vomeronasal organ of conspecifics. PMID- 11058764 TI - Mammalian odorant binding proteins. AB - Odorant binding proteins (OBPs) pertain to one of the most abundant classes of proteins found in the olfactory apparatus. OBPs are a sub-class of lipocalins, defined by their property of reversibly binding volatile chemicals, that we call 'odorants'. Numerous sequences of OBPs are now available, derived from protein sequencing from nasal mucus material, or from DNA sequences. The structural knowledge of OBPs has been improved too in recent years, with the availability of two X-ray structures. The physiological role of OBPs remains, however, essentially hypothetical, and most probably, not linked to a function of odor transport. The present knowledge on OBP biochemistry, sequence and structure will be examined here in relation to the different functional hypotheses proposed for OBPs. PMID- 11058765 TI - Human tear lipocalin. AB - Human tear prealbumin, now called tear lipocalin, was originally described as a major protein of human tear fluid, which was thought to be tear specific. However, recent investigations demonstrated that it is identical with lingual von Ebner's gland protein, and is also produced in prostate, nasal mucosa and tracheal mucosa. Homologous proteins have been found in rat, pig and probably dog and horse. Tear lipocalin is an unusual lipocalin member, because of its high promiscuity for relative insoluble lipids and binding characteristics that differ from other members. In addition, it shows inhibitory activity on cysteine proteinases similar to cystatins, a feature unique among lipocalins. Although it acts as the principal lipid binding protein in tear fluid, a more general physiological function has to be proposed due to its wide distribution and properties. It would be ideally suited for scavenging of lipophilic, potentially harmful substances and thus might act as a general protection factor of epithelia. PMID- 11058766 TI - Rat probasin: structure and function of an outlier lipocalin. AB - Probasin (PB) occurs both as a secreted and a nuclear protein that is abundantly expressed in the epithelial cells of the rat prostate. A genomic clone of 17.5 kb gene was isolated from a rat liver genomic library, determining that the probasin gene was comprised of seven exons where the splice donor/acceptor sites conformed to the GT/AG consensus sequence. The exon number and size are remarkably similar to those of aphrodisin, rat alpha(2)-urinary globulin and major urinary protein, outlier members of the lipocalin superfamily. In addition, alignment of the deduced amino acids determined that the probasin gene also contains the glycine-X tryptophan (G-X-W) motif similar to that of human retinol serum binding protein which binds retinol, and the C-X-X-X-C motif also found in insect lipocalins that bind pheromones. The cysteine residues in exons 3 and 6 are conserved, predicting a secondary structure of eight beta-sheets and the alpha-helix commonly seen in the lipocalin superfamily. Unique PB characteristics include a large genomic fragment (17.5 kb compared to the 3-5 kb seen in other lipocalin genes) and an isoelectric point (pI) of 11.5 which is very basic compared to that of the other more acidic lipocalins. Functionally, PB gene expression is regulated by androgens and zinc in the epithelial cells of the rodent prostate. The 5' flanking region of probasin contains two androgen receptor binding sites that allow androgen-specific gene expression as well as prostate-specific elements that target and maintain high levels of transgene expression in several PB transgenic mouse models. PMID- 11058767 TI - Biochemical, structural, genetic, physiological, and pathophysiological features of lipocalin-type prostaglandin D synthase. AB - Lipocalin-type prostaglandin (PG) D synthase (PGDS) catalyzes the isomerization of PGH(2), a common precursor of various prostanoids, to produce PGD(2), a potent endogenous somnogen and nociceptive modulator, in the presence of sulfhydryl compounds. PGDS is an N-glycosylated monomeric protein with an M(r) of 20000 31000 depending on the size of the glycosyl moiety. PGDS is localized in the central nervous system and male genital organs of various mammals and in the human heart and is secreted into the cerebrospinal fluid, seminal plasma, and plasma, respectively, as beta-trace. The PGDS concentrations in these body fluids are useful for the diagnosis of several neurological disorders, dysfunction of sperm formation, and cardiovascular and renal diseases. The cDNA and gene for PGDS have been isolated from several animal species, and the tissue distribution and cellular localization have also been determined. This enzyme is considered to be a dual functional protein; i.e. it acts as a PGD(2)-producing enzyme and also as a lipophilic ligand-binding protein, because the enzyme binds biliverdin, bilirubin (K(d)=30 nM), retinaldehyde, retinoic acid (K(d)=80 nM) with high affinities. X-ray crystallographic analyses revealed that PGDS possesses a beta barrel structure with a hydrophobic pocket in which an active thiol, Cys(65), the active center for the catalytic reaction, was located facing to the inside of the pocket. Gene-knockout and transgenic mice for PGDS were generated and found to have abnormalities in the regulation of nociception and sleep. PMID- 11058768 TI - Human neutrophil gelatinase-associated lipocalin and homologous proteins in rat and mouse. AB - Neutrophil gelatinase-associated lipocalin (NGAL) is a 25-kDa lipocalin originally purified from human neutrophils. It exists in monomeric and homo- and heterodimeric forms, the latter as a dimer with human neutrophil gelatinase. It is secreted from specific granules of activated human neutrophils. Homologous proteins have been identified in mouse (24p3/uterocalin) and rat (alpha(2) microglobulin-related protein/neu-related lipocalin). Structural data have confirmed a typical lipocalin fold of NGAL with an eight-stranded beta-barrel, but with an unusually large cavity lined with more polar and positively charged amino acid residues than normally seen in lipocalins. Chemotactic formyl-peptides from bacteria have been proposed as ligands of NGAL, but binding experiments and the structure of NGAL do not support this hypothesis. Besides neutrophils, NGAL is expressed in most tissues normally exposed to microorganisms, and its synthesis is induced in epithelial cells during inflammation. This may indicate either a microbicidal activity of NGAL or a role in regulation of inflammation or cellular growth, putative functions yet to be demonstrated. PMID- 11058769 TI - Immunocalins: a lipocalin subfamily that modulates immune and inflammatory responses. AB - A subset of the lipocalins, notably alpha(1)-acid glycoprotein, alpha(1) microglobulin, and glycodelin, exert significant immunomodulatory effects in vitro. Interestingly, all three are encoded from the q32-34 region of human chromosome 9, together with at least four other lipocalins (neutrophil gelatinase associated lipocalin, complement factor gamma-subunit, tear prealbumin, and prostaglandin D synthase) that also may have anti-inflammatory and/or antimicrobial activity. This review addresses important features of this genetically linked subfamily of lipocalins (involvement with the acute phase response, immunomodulatory and anti-inflammatory properties, the tissue localization, complex formation with other proteins and receptors, etc.). It is likely that these proteins have evolved to be an integrated part of the body's defense system as part of the extended cytokine network. Its members exert a regulatory, dampening influence on the inflammatory cascade, thereby protecting against tissue damage from excessive inflammation. That most major mammalian allergens are lipocalins may reflect this connection of lipocalins with the immune system. We propose that this immunologically active lipocalin subset be named the 'immunocalins', signifying not only the structural homology and close genetic linkage of its members, but also their protective involvement with immunological and inflammatory processes. As immune mediators, immunocalins appear to use at least three interactive sites: the lipocalin 'pocket', binding sites for other plasma proteins, and binding sites for cell surface receptors. PMID- 11058770 TI - Lipocalins as biochemical markers of disease. AB - Lipocalins as biochemical markers of disease have been used extensively. The clinical indications relate to almost any field of medicine, such as inflammatory disease, cancer, lipid disorders, liver and kidney function. Some of the more well-known lipocalins that have been used as markers of disease are orosomucoid, Protein HC (alpha(1)-microglobulin), apolipoprotein D, retinol-binding protein, complement C8 gamma, prostaglandin D synthase and human tear prealbumin, and these markers will be briefly reviewed in this article. Emphasis, however, will be put on the description of another newly described lipocalin, i.e. human neutrophil lipocalin/neutrophil gelatinase-associated lipocalin (HNL/NGAL), since the body fluid measurement of HNL/NGAL was shown to be a superior means to distinguish between acute viral and bacterial infections and also to accurately reflect the activity and involvement of neutrophils in a variety of other diseases. PMID- 11058771 TI - Lipocalins as allergens. AB - The term allergy refers to clinical conditions caused by an inappropriate immune response to innocuous proteins in genetically predisposed persons. Allergens of animal origin are responsible for a significant proportion of allergies. In recent years, it has become evident that practically all respiratory animal allergens characterized at the molecular level belong to the lipocalin family of proteins. The current list comprises the major allergens of horse, cow, dog, mouse, rat and cockroach as well as beta-lactoglobulin of cow's milk. While the molecular structure of all these allergens is known, far less information is available regarding their immunological characteristics. Knowing the way the immune system recognizes these allergens and reacts to them might, however, be the key for discovering the common denominator of the allergenicity of lipocalins. The human body contains numerous endogenous lipocalins, and the immune system has to adapt to their presence. We have proposed that under these conditions the immune response against the lipocalin allergens which are structurally related to endogenous lipocalins might be the pathway to allergy in genetically predisposed persons. The same might well apply also to other allergens with homologous endogenous counterparts. PMID- 11058772 TI - Lipocalins and cancer. AB - Lipocalins are mainly extracellular carriers of lipophilic molecules, though exceptions with properties like prostaglandin synthesis and protease inhibition are observed for specific lipocalins. The interest concerning lipocalins in cancer has so far been focussed to the variations in concentration and the modification of lipocalin expression in distinct cancer forms. In addition, lipocalins have been assigned a role in cell regulation. The influence of the extracellular lipocalins on intracellular cell regulation events is not fully understood, but several of the lipocalin ligands are also well-known agents in cell differentiation and proliferation. Lipophilic ligands can, after lipocalin mediated transport to the cell surface, penetrate the cell membrane and interact with proteins in the cytosol and/or the nucleus. The signaling routes of the lipocalin ligands, retinoids and fatty acids are presented and discussed. Tumor growth in tissue is restricted by extracellular protease/protease inhibitor interactions. Several lipocalins also have protease inhibitory properties and possess the ability to interact with tumor specific proteases, revealing another pathway for lipocalins to interact with cancer cells. PMID- 11058773 TI - Beyond the superfamily: the lipocalin receptors. AB - Lipocalins are characterized by multiple molecular recognition properties including the ability to bind to cell surface receptors. Receptors for a number of lipocalins have been identified. These include receptors for alpha-1 microglobulin, insecticyanin, glycodelin, retinol-binding protein, alpha-1-acid glycoprotein, beta-lactoglobulin and odorant-binding protein. The properties of these receptors are summarized and discussed. PMID- 11058774 TI - Lipocalins as a scaffold. AB - The concept of scaffolds that can be equipped with artificial biochemically active sites has gained recent interest in the field of protein design. Members of the lipocalin protein family represent promising model systems in this respect. Especially prototypic lipocalins, such as the retinol-binding protein or the bilin-binding protein (BBP), exhibit a structurally simple one-domain fold with a conformationally well conserved beta-barrel as their central motif. This type of supersecondary structure is made of a cylindrically closed beta-sheet of eight antiparallel strands. At the open end of the barrel the beta-strands are connected by four loops in a pairwise manner so that a pocket for the ligand is formed. In a rational protein design study a metal-binding site was functionally grafted on the solvent-exposed surface of the beta-barrel, whereby the rigid backbone conformation permitted the spatially defined arrangement of three His side chains. In a combinatorial protein design approach, the natural ligand pocket of a lipocalin was reshaped. In this manner variants of the BBP were engineered which exhibit high affinity and remarkable specificity for haptens like fluorescein and digoxigenin. The so-called 'anticalins', i.e. artificial lipocalins recognizing prescribed ligands, could provide an interesting alternative to recombinant antibody fragments. Consequently, the use of lipocalins as a scaffold opens new applications for members of this functionally diverse protein family in biotechnology and medicine. PMID- 11058775 TI - The lipocalin website. AB - We introduce a website devoted to the lipocalins. The website contains data on lipocalin structures and sequences, as well as reviewing lipocalin biology and biochemistry. Our hope is that it can act as a focus for future research into the lipocalin protein family. The website can be accessed at the following URL: http://www. jenner.ac.uk/lipocalin.htm. PMID- 11058776 TI - Using a Bayesian belief network to aid differential diagnosis of tropical bovine diseases. AB - The examination of presenting signs has always played an important role in the diagnosis of diseases in animal populations. In the case of diseases of tropical cattle, such expertise is often scarce and confined to those experts with many years of experience. To capture, conserve and disseminate such valuable expert knowledge remains a key challenge to the application of knowledge-based systems in veterinary medicine. In this communication, we explore the use of a Bayesian belief network to quantify expert opinion with a view to estimating the likelihood of various diseases in the presence and absence of certain signs. Information was elicited from a panel of 44 experienced veterinarians to provide the response matrix of 27 signs associated with 20 commonly occurring diseases in sub-Saharan cattle. Using this prior information, estimates of the probability of certain signs occurring with each disease were calculated from which the Bayesian belief network was able to propagate the posterior probability of each of the diseases based on the observed signs. The method as an aid in making diagnosis is discussed. It is recognised that such an approach is but one strand in the process of arriving at a diagnosis. For ease of use and accessibility, the approach has been converted into the software program CaDDiS (Cattle Disease Diagnosis System) which is available for consultation on the World Wide Web. PMID- 11058777 TI - Operation-management factors associated with early-postnatal mortality of US foals. AB - Of 7320 equine foals reported born alive during 1997 on 1043 operations that had equids on 1 January 1997, and that participated in the United States National Animal Health Monitoring System (NAHMS) Equine 1998 Study, 120 foals were reported to have died (by either euthanasia or natural causes) within the first 2 days of a live birth. The weighted estimate was 1.7% mortality (standard error=0.5) within the first 2 days of live birth for all foals born on operations in the 28 states included in the study.A multivariable logistic-regression model revealed that foals born in the southern region were more likely to have been reported to have died within the first 2 days of live birth than in the western region. In addition, the following operation-level factors were associated with increased odds of a foal dying within the first 2 days of live birth: not routinely testing newborn foals for adequate absorption of colostral immunoglobulins during the first 2 days of life; adding new resident equids to the operation during 1997; having non-resident equids stay on the operation for 1 30 days during 1997; never requiring an official health certificate (for operations that had non-resident equids stay on the operation for 1-30 days); using something other than straw or hay as the predominant bedding type; and feeding equids a vitamin-mineral supplement/premix with forage and/or grain. PMID- 11058778 TI - Response of Bali cattle (Bos javanicus) to vaccination with Brucella abortus strain 19 in West Timor. AB - A trial was conducted in two villages (one containing cattle infected with brucellosis and one not containing infected cattle) in Timor, Indonesia to determine the serological response to vaccination with Brucella abortus strain 19 in Bali cattle (Bos javanicus) (n = 599). Mature female cattle were immunised with low-dose strain 19 (2x10(8)-6x10(8) colony forming units) and calves (6-12 months) with high-dose strain 19 (4x10(10)-12x10(10) colony forming units). Other mature females and calves were inoculated with sterile vaccine diluent and formed a non-vaccinated in-contact control group. The seroprevalence and mean titres were highest in the vaccinated cattle 3 months after vaccination. These then receded, however, 1% of vaccinated calves and 1.9% of vaccinated cows from the village without infected cattle were still seropositive on the complement fixation test (CFT) 24 months after vaccination. Non-vaccinated seropositive animals were more likely to have aborted or had a stillbirth and were less likely to have produced a calf than were seronegative cows from the village containing infected animals. We concluded that strain 19 vaccine induced protection in Bali cattle and that this vaccine might play an important role in the control of bovine brucellosis in Timor. PMID- 11058779 TI - Spatial and temporal comparison of selected cancers in dogs and humans, Michigan, USA, 1964-1994. AB - Our aim was to investigate the geographic and time distributions of some biologically similar neoplasms in dogs and humans living in Michigan, USA, between 1964 and 1994. Our objective was to describe and compare the patterns of cancer in the two species while assessing the strength and dependence of those patterns. In this retrospective, registry-based study, histologically confirmed incident human and canine cancer cases were mapped, and second-order (K function) spatial analysis and one-dimensional nearest neighbor temporal analysis were performed on residence addresses and dates of hospital discharge/diagnosis. Included in the study were all 528 incident cases of canine lymphosarcoma, mammary adenocarcinoma, melanoma and spindle-cell sarcomas diagnosed at a veterinary teaching hospital between 1964 and 1994 having residence addresses in Ingham, Oakland, and Wayne Counties; and a stratified random sample of 913 incident human cases of comparable cancers diagnosed during the same time period from the same counties. Results suggest that processes determining spatial aggregation of cases in dogs and humans were not independent of each other, did not act uniformly over different geographic areas, operated at spatial scales <2000 m regardless of species, and tend to act upon dogs more strongly at shorter distances than on humans. Little evidence of interspecies concurrence of temporal clustering was found. PMID- 11058780 TI - Age and seasonal variations in the prevalence of Oestrus ovis larvae among sheep in northern Jordan. AB - During the period March 1996-July 1997, 417 heads of Awassi sheep slaughtered at the Irbid Abattoir (northern Jordan) were examined for the three larval instars (L1, L2 and L3) of Oestrus ovis. Of the 417 heads, 242 (58%) were infested with O. ovis larvae. Larval numbers were highly aggregated. The lowest number of larvae and the lower quartile were both zero, whilst the median was two and the upper quartile was 12. The highest number of larvae recovered from one head was 151. All three larval instars were observed in each month of the year. July and October had the highest proportions of L1, 75 and 78%, respectively, among infected animals (adjusted for age). The number of larvae increased with age. Infestation with live larvae was associated with inflammatory responses in the upper respiratory tract and with catarrhal or purulent discharge. The percentage of infested sheep and the mean monthly total number of larvae/sheep peaked in the warmer part of the year. Most larvae were L1 except during the spring when L2 and L3 predominated. Distribution analysis demonstrates that the numbers of larvae recovered in the sheep population followed a negative-binomial distribution. Furthermore, the negative-binomial constant k for each month correlated with the monthly prevalence. PMID- 11058781 TI - Preventing the spread of maedi-visna in sheep through a voluntary control programme in Finland. AB - The sheep disease maedi-visna (MV) was introduced into Finland in 1981 and had spread to eight flocks in the southwestern part of the country when first detected in a survey in 1994. Six more seropositive flocks were subsequently traced, bringing the total to 14. MV has a notifiable disease status in Finland that provides for official restrictive measures to which all infected herds are subject. These measures are withdrawn once the seropositive animals and their progeny are culled and the flock has showed negative signs in the test done twice, or after total culling. A voluntary control programme was initiated in January 1995 to extend official control efforts. The programme furnishes a guideline for culling, restrictions on contacts, and a timetable for testing the flock to attain MV-free status. Seven flocks of the 14 were slaughtered either immediately or after a period under restrictive measures. One flock finished sheep production after four years under restrictive measures. Selective culling and repeated testing was attempted with the other six flocks, three of which attained MV-free status. One flock finished sheep production after two years in the control programme, the other two dropped out of the programme when the restrictive measures were withdrawn. It was concluded that the control programme was salient in eradicating MV from Finland and that serological monitoring of the situation must be continuous. PMID- 11058782 TI - Genetic studies: look no further than your own backyard. PMID- 11058783 TI - Breeding of Escherichia coli based on colour. PMID- 11058784 TI - Biocavity lasers for biomedicine. AB - Laser technology has advanced dramatically and is an integral part of the healthcare delivery systems of today. Lasers are used in laboratory analyses of human blood samples and serve as surgical tools that kill, burn or cut tissue. Recent semiconductor microtechnology has reduced the laser size to the size of a biological cell or even a virus particle. The integration of these ultra-small lasers with biological systems makes it possible to create microelectrical mechanical systems that might revolutionize healthcare delivery. PMID- 11058785 TI - Frontiers in chemical genetics. AB - New methods enable the identification of compounds that both induce a specific cellular state and lead to identification of proteins that regulate that state. Together, developments in three critical areas: chemical diversity, phenotype based screening and target identification, enable the systematic application of this chemical genetic approach to almost any biological problem or disease process. PMID- 11058786 TI - New perspectives on the design of cytokines and growth factors. AB - A combination of molecular modelling, conventional epitope scanning and combinatorial techniques, such as phage display and DNA shuffling, has greatly improved our understanding of ligand-receptor interactions. It has therefore been possible to develop powerful cytokine-growth factor antagonists and new designer cytokines, with altered receptor specificities or with greatly enhanced biological activity. Recently, small circular peptides that mimic or block the effects of natural cytokines and growth factors have been developed; such small peptides are likely to open new avenues in therapeutics. PMID- 11058787 TI - Allosterically controllable maxizymes cleave mRNA with high efficiency and specificity. AB - Ribozymes are small and versatile nucleic acids that can cleave RNA molecules at specific sites. However, because of the limited number of cleavable sequences on the target mRNA, in some cases conventional ribozymes do not have precise cleavage specificity. To overcome this problem, an allosteric version (a maxizyme) was developed that displayed activity and specificity in vivo. More than five custom-designed maxizymes have demonstrated sensor functions, which indicates that the technology might be broadly applicable in molecular biology and possibly in the clinic. PMID- 11058788 TI - Bioencapsulation within synthetic polymers (Part 2): non-sol-gel protein-polymer biocomposites. AB - Since the introduction of sol-gel bioencapsulation and the demonstration that biological function can be incorporated into, and preserved within, polymer matrices, a number of alternative polymers have been used to immobilize proteins. Various enzymes have been trapped in such diverse polymers as epoxy-amine resins, polyvinyl plastics, polyurethane foams and silicone elastomers. Together with sol gel encapsulates, these biocomposites represent a powerful approach for immobilizing biological materials for applications as biosensors and biocatalysts, and hold promise as bioactive, fouling-resistant polymers for environmental, food and medical uses. Although still at the developmental stage, these biocomposites promise to revolutionize the whole arena of high-performance bioimmobilization. PMID- 11058789 TI - Polymorphisms in the alpha-2 macroglobulin gene in psychogeriatric patients. AB - Recent reports have suggested that genetic polymorphisms in the alpha-2 macroglobulin (A2M) gene are associated with an increased risk for Alzheimer's disease. In the present study we tested two polymorphisms in the alpha-2 macroglobulin gene, a 5 bp deletion at the 5' splice site of exon 18 and a G/A point mutation (V1000I) in exon 24, in a sample of 118 healthy, non demented controls and 238 consecutively recruited gerontopsychiatric patients, diagnosed as: Alzheimer's disease (N=88), mild cognitive impairment (N=32), subjective cognitive complaints (N=54) and depression/other psychiatric disorders (N=64). The aim of this study was to test whether the investigated polymorphisms has a high enough selectivity and specificity to distinguish between the different gerontopsychiatric disorders or to differentiate genetically AD from other forms of dementia, respectively. Also a possible relation to the APOE varepsilon4 polymorphism was examined. Our study failed to show an association between the two investigated polymorphisms in the alpha-2 macroglobulin gene and any of the four different psychogeriatric patient subgroups, either alone or in combination with the APOE varepsilon4 genotype. PMID- 11058790 TI - Effect of retinoic acid and ethanol on retinoic acid receptor beta and glial fibrillary acidic protein mRNA expression in human astrocytoma cells. AB - This work explores the hypothesis that perturbations caused by ethanol on the regulatory role of retinoids in brain development may be a mechanism involved in the neuropathology of fetal alcohol syndrome. The interaction of ethanol and retinoic acid (RA) on RA receptor (RAR) beta and glial fibrillary acidic protein (GFAP) mRNA expression is evaluated. In the U-373 MG astrocytoma, mRNA expression of RAR beta was increased and GFAP was decreased by RA. Ethanol decreased the expression of RAR beta mRNA, but increased that of GFAP. The RA-stimulated increase in RAR beta was not affected by the presence of ethanol. RA prevented the ethanol-induced increase in GFAP mRNA. Cycloheximide abolished only the GFAP response to ethanol. This work shows that an interrelationship between ethanol and RA exists in the astrocyte. PMID- 11058791 TI - Abnormal synaptic organization between granule cells and Purkinje cells in the new ataxic mutant mouse, pogo. AB - The pogo mouse is a new ataxic mutant derived from the Korean wild mouse. The pathological manifestations include difficulty in maintaining normal posture and the inability to walk straight. The ataxia becomes apparent at about 2 weeks of age. Electron microscopic studies of the pogo/pogo homozygous cerebellum, revealed that the ectopic spines emanating from the primary dendrite of Purkinje cells were observed. Major difference between pogo/pogo homozygous and non affected pogo/+ heterozygous was the synaptic organization of the molecular layer. Parallel fiber varicosities were larger than normal and a single fiber often established synaptic contacts with up to four dendritic spines of a Purkinje cell. This correlation between the presence of altered synaptic organization in the cerebellum and ataxia in pogo/pogo mutant mice warrants further investigation. PMID- 11058792 TI - Phosphofructokinase, a glycolytic regulatory enzyme has a crucial role for maintenance of synaptic activity in guinea pig hippocampal slices. AB - To clarify the importance of glucose metabolism in maintaining neural activity, we investigated the effects of lowering the concentration of glucose in perfusion medium on synaptic activity (population spikes, PS) and on the level of high energy phosphates in the region of dentate gyrus (DG), CA3 and CA1 area of hippocampal slices of guinea pig. Further we determined the activity of phosphofructokinase (PFK) and hexokinase (HK) of these areas. Lowering the concentration of glucose from 10 to 5, 4, 3, 1 and 0 (mM) caused a reduction in the PS amplitude. The PS in CA3 and CA1 region decayed and extinguished much faster than that in DG area. The ATP and CrP levels in these three areas were well preserved even after lowering the concentration of glucose to 3mM, and the patterns of reduction showed no difference in these three areas. On the other hand, there were significant differences in the activity of PFK in the DG, CA3 and CA1 areas, in contrast with HK which showed no significant differences. PFK activity was highest in the DG area and followed by CA3 and CA1 regions. The sensitivity in maintaining synaptic activity to lowered glucose concentrations showed good parallels with the activity of PFK in these regions, indicating that non-oxidative glycolytic metabolism regulated by PFK is crucial for the maintenance of synaptic activity PMID- 11058793 TI - The translating brain: cerebral activation patterns during simultaneous interpreting. AB - Brain activation was measured in professional interpreters during simultaneous interpreting (SI) vs. repetition (shadowing) of auditorily presented text by positron emission tomography (PET). SI into the native language (Finnish) elicited left frontal activation increases. SI into the non-native language (English) elicited much more extensive left-sided fronto-temporal activation increases. Our results indicate that SI activates predominantly left-hemispheric structures (particularly the left dorsolateral frontal cortex) previously related to lexical search, semantic processing and verbal working memory. Brain activation patterns were clearly modulated by direction of translation, with more extensive activation during translation into the non-native language which is often considered to a be more demanding task. PMID- 11058794 TI - The hemispherical laterality of the visual evoked potentials during simple dot stimulus in normal human subjects. AB - We studied scalp visual evoked potentials (VEPs) during a simple visual attention paradigm using a dot stimulus, which was presented every 1600 ms, at the center of a screen. The visual attention paradigm consisted of three tasks: task R, task L and task N. The subjects were instructed to press a button either with the right hand (task R) or with the left hand (task L) after seeing the dot. They were also instructed just to look at a dot, without pressing the button (task N). We defined N1 as a negative wave with a latency of 50-130 ms, P1 as a positive wave with a latency of 110-150 ms and N2 as a negative wave with a latency of 130 210 ms. During task R, P1-N2 amplitude at T6 was significantly greater than that at T5. The N1-P1 and N2 amplitudes at O2 were significantly greater than those at O1. During task L, the waveforms at T6 and O2 were more clearly detected than those at T5 and O1. We conclude that there is a functional dominance of the right cerebral hemisphere in our simple visual reaction tasks. PMID- 11058795 TI - Gene expression of alpha(1A)-adrenoceptor but not alpha(1B)-adrenoceptor in cultured myoblast C(2)C(12) cells of mice. AB - Subtypes of alpha(1)-adrenoceptor (alpha(1)-AR) in the cultured myoblast C(2)C(12) cells have been examined using molecular biological identifications. Expression of the two distinct mRNAs that encode proteins of alpha(1A)- and alpha(1B)-AR was studied using reverse transcription combined with polymerase chain reaction (RT-PCR). Results of RT-PCR demonstrated a marked expression of alpha(1A)-AR in the prostate of rats. Samples from the C(2)C(12) cells under the same amount of amplification showed the expression of alpha(1A)-AR at a level slightly lower than that from the prostate of rats. Western blotting analysis using receptor subtype-specific antibody also indicated that the alpha(1A)-AR was expressed in C(2)C(12) cells as well as in prostate of rats. However, although the expression of alpha(1B)-AR was obtained in the spleen of rats, expression of alpha(1B)-AR was undetectable in the C(2)C(12) cells either the results of RT-PCR or the Western blotting analysis. The present study suggests that alpha(1A)-AR is a major subtype of alpha(1)-AR in the C(2)C(12) cell line. PMID- 11058796 TI - Ionotropic glutamate receptors expressed in human retinoblastoma Y79 cells. AB - Mammalian retinal photoreceptors and pinealocytes have common characteristic that they secrete melatonin and L-glutamate as chemical transmitters. Although pinealocytes express glutamate receptors and receive glutamate signals, whether or not photoreceptors express glutamate receptors is unknown. Here, we investigated the expression of the glutamate receptors in cultured Y79 clonal human retinoblastoma cells, as model systems of photoreceptors. Reverse transcription-polymerase chain reaction (RT-PCR) analysis indicated that GluR1, GluR5, GluR7, EAA2, NR1, NR2A and NR2D mRNAs were present in the cultured cells. Northern analysis confirmed the presence of GluR7, EAA2, NR1, NR2A and NR2D mRNAs, while other mRNAs were under the detection limit. Addition of (RS)-alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid or kainate increases intracellular (Ca(2+)) in Fura-2 loaded cells, which is blocked by 6-cyano-7 nitroquinoxaline-2,3-dione. N-methyl-D-aspartate also increases intracellular (Ca(2+)). These results demonstrated the presence of functional ionotropic receptors in Y79 cells. PMID- 11058797 TI - Altered phase relation between sleep timing and core body temperature rhythm in delayed sleep phase syndrome and non-24-hour sleep-wake syndrome in humans. AB - Changes in the phase relation between sleep timing and the circadian pacemaker are suspected to have an etiological significance in circadian rhythm sleep disorders. Simultaneous recordings of rest-activity and rectal temperature in seven sighted delayed sleep phase syndrome (DSPS) patients, seven sighted non-24 h sleep-wake syndrome (non-24) patients, and 14 healthy controls were made for 10 14 days continuously in the subjects' homes. We found that sleep length and the interval from the body temperature (BT) trough to sleep offset were significantly longer in both non-24 and DSPS patients than in the controls, and that the interval between sleep onset and the BT trough was significantly less in the non 24 patients than in the DSPS patients and the controls. We postulate these alterations in phase relation to be associated with phase changes of the circadian pacemaker via different illumination timings. PMID- 11058798 TI - US3 protein kinase of herpes simplex virus protects primary afferent neurons from virus-induced apoptosis in ICR mice. AB - Possible roles of the US3 gene of the herpes simplex virus (HSV) in the interaction between the virus and primary afferent neurons were examined. Neuronal apoptosis was observed in the trigeminal ganglion of mice that were infected with the wild-type (wt) of HSV-2 strain 186 and with US3-deficient mutant virus (L1BR1). In wt virus-infected mice, many HSV-immunoreactive (HSV-ir) cells were seen throughout the trigeminal ganglion, although no apoptotic change was detected. On the other hand, HSV-ir cells in L1BR1-infected mice were found only in the ophthalmic division of the trigeminal ganglions. Examination by HSV immunohistochemistry combined with the terminal deoxynucleotidal transferase (Tdt)-mediated deoxyuridin 5'-triphosphate (dUTP) nick-end labeling (TUNEL) method showed that DNA fragmentation had occurred in almost all HSV-ir cells in the L1BRI-infected ganglion. Ultrastructurally, many viral particles were detected in apoptotic ganglionic neurons of mice infected with L1BR1. These results indicate that US3 protein kinase (US3pk) played a role in protecting HSV infected primary afferent neurons from apoptotic cell death. The present study suggests that US3pk plays a role when HSV establishes latent infections in the sensory ganglia. PMID- 11058799 TI - Effects of age on cholinergic vasodilation of cortical cerebral blood vessels in rats. AB - The present study examined the age-related changes in the cholinergic vasodilative system originating in the nucleus basalis of Meynert (NBM) and projecting to the cerebral cortex using Wistar rats of three different ages; young adult (4-7 months), old (24-25 months), and very old (32-42 months) rats. The vasodilative responses in frontal and parietal cortices, measured by laser Doppler flowmetry, induced by electrical stimulation of NBM without blood pressure response were well maintained in old rats, but declined significantly in very old rats. Extracellular acethylcholine (ACh) release in both cortices collected by a microdialysis technique showed both basal levels and response to NBM stimulation to be well maintained in both old and very old rats. The vasodilative cerebral blood flow response elicited by stimulation of the muscarinic ACh receptors, using their agonist, arecoline, was also well maintained in old and very old rats. Considering the present data and our previous finding that the cerebral cortical vasodilative response to activation of the nicotinic ACh receptors using their agonist, nicotine, was markedly reduced in very old rats (Neurosci. Lett., 228 (1997) 203), it was concluded that the age-related decline of nicotinic ACh receptor activity was a cause of the decline of the vasodilative responses elicited by NBM stimulation in very old rats. This result suggests that a reduction of the cholinergic vasodilative system in very old rats due to decreased activity of the nicotinic ACh receptor may cause insufficient blood flow in the cortex when the cortical neurons require. PMID- 11058800 TI - Synaptic communication of cellular oscillations in the rat suprachiasmatic neurons. AB - Circadian firing rhythms of cultured rat suprachiasmatic nucleus were measured simultaneously from 4-8 neurons by using a multi-electrode dish and neuronal interactions were examined by a cross-correlation analysis of spontaneous action potentials. Functional connections were detected in the neuron pairs showing synchronized circadian firing rhythms, and when the connections were lost, firing rhythms were desynchronized. After the prolonged treatment with tetrodotoxin, cross-correlation and circadian rhythm synchronization were abolished concomitantly in most neuron pairs. Cellular mechanisms involving Na(+)-channel dependent communication are responsible for the synchronization of the circadian rhythms in individual suprachiasmatic nucleus (SCN) neurons. PMID- 11058801 TI - Phosphorylation of c-Jun NH(2)-terminal kinase and p38 mitogen-activated protein kinase after transient forebrain ischemia in mice. AB - We investigated the activation of c-Jun NH(2)-terminal kinase (JNK) and p38 mitogen-activated protein kinase (MAPK) during transient forebrain ischemia to clarify the roles of these stress kinases during brain ischemia. Mice were subjected bilateral common carotid artery (BCCA) occlusion for 20 min followed by reperfusion. Immunohistochemical analysis and Western blot analysis for active JNK and active p38 MAPK were performed at 0, 5, 10, 30 and 150 min after reperfusion. After 5 min of reperfusion, active JNK and p38 MAPK immunoreactivities were enhanced in neurons in the cerebral cortex and hippocampus; this activation peaked at 30 min of reperfusion. Stress kinases activation dominantly occurred in the similar regions, in which neurons with fragmented DNA were detected at 72 h after reperfusion. Western blot analysis indicated that JNK 1, JNK 2 and p38 MAPK were activated at 10 and 30 min after reperfusion. These findings indicate that JNK and p38 MAPK pathways may play important roles in neuronal death during brain ischemia. PMID- 11058802 TI - Isolation and purification of an epidermal growth factor receptor-related inhibitor of cell growth from cultured rat astrocytes. AB - Astrocytes rarely undergo mitotic cell division in the developed adult. However, following injury to the central nervous system (CNS), astrocytes undergo rapid cell division and reactive astrocytes encapsulate exposed areas of the CNS producing a glial scar. Studies indicate the existence of a family of epidermal growth factor receptor (EGFr) related glycosidic mitogen inhibitors with growth inhibiting properties for astrocytes. Here, for the first time, we describe the isolation of such a factor from cultured rat astrocytes by ion-exchange and immunoaffinity chromatography. The isolated factor is glycosidic in nature, cross reacts with the anti-EGFr monoclonal antibody Clone 29.1, and inhibits the growth of primary astrocytes and N2a neuroblastoma cells in vitro. PMID- 11058803 TI - Electrochemical detection of nitroso-arginine as an intermediate between N hydroxy-arginine and citrulline. An in vitro versus in vivo study using microcarbon electrodes in neuronal nitric oxide synthase and mice brain. AB - The aim of the study was to describe in vivo and in vitro the transformation of N hydroxy-arginine (NHA) into nitrite and citrulline. The products of NHA oxidation were studied by electrochemical methods. Cyclic voltammetry of NHA on microcarbon electrode showed an oxidation in two steps with one electron and one proton exchanged at each step. The first step gave a radical species NHA(.) with a half life shorter than 1 micros and the second step gave nitroso-arginine (NA) with a half-life of about 1 s (1.5 s). Coulometric oxidation of NHA gave citrulline and nitrite. Differential pulse voltammetry (DPV) in vivo and in vitro gave a peak in reduction at -1.66 V vs Ag/AgCl for NA. After reductive adsorption of NA on the microelectrode surface in mice brain it gave the two peaks of NHA in oxidation plus another peak identified as nitrite. DPV in native and recombinant rat brain nitric oxide (NO)-synthase gave NA signal permitting K(m) and V(max) determination. All these results showed that NA was synthetized by NO-synthases before the final products, citrulline and nitrite. PMID- 11058804 TI - Bradykinin activates phospholipase D2 via protein kinase cdelta in PC12 cells. AB - Bradykinin (BK) activates phospholipase D (PLD) and induces several responses such as catecholamine secretion, collapse of growth cones, and gene expression in PC12 pheochromocytoma cells. Although two distinct PLD isozymes, PLD1 and PLD2, have been cloned from mammalian cells, the regulatory mechanism for each PLD isozyme by BK is not clear. In our present study, we investigated the activation mechanism of PLD2 by BK in PLD2-overexpressing PC12 cells. BK stimulated PLD2 activity in a concentration-dependent manner within 1 min and this activation was inhibited by pretreatment of the cells with protein kinase C (PKC) inhibitor. PKCalpha and PKCdelta translocated from cytosol to membrane upon BK treatment, and rottlerin potently inhibited the activation of PLD2 by BK. These results suggest that BK activates PLD2 via PKCdelta in PC12 cells. PMID- 11058805 TI - Skeletal drug delivery systems. AB - Selective drug delivery to any organ becomes very important in certain diseases and clinical manifestations, especially when the drug affects other exposed tissues adversely. The importance of selective drug action is still further increased when the affected part is poorly perfused. Although a network of blood vessels is present throughout skeletal tissue (or bone), it is not sufficient for immediate delivery of drugs to the desired site of action in the tissue and in sufficient amounts over appropriate time periods. Hence, selective drug delivery to the skeletal system has remained a great challenge to pharmaceutical scientists over the years. However, in the recent past, attention has been focused on the importance of skeletal drug delivery and the first Skeletal Drug Delivery System (SDDS) was introduced by Bucholz and Engelbrecht in 1970 for the delivery of drugs to skeletal tissues at a high concentration to achieve desirable therapeutic effects. SDDS is used to deliver the drug directly to skeletal tissue through self-setting cement, which also acts as a bone filler, thereby improving the therapeutic effectiveness of drugs in bone diseases. The present review highlights the applicability of various materials as bone fillers for the purpose of skeletal drug delivery. PMID- 11058806 TI - Solid dispersion of ketoprofen in pellets. AB - The formulation of ternary solid dispersions of ketoprofen with Macrogol and kollagen hydrolizate derivative as carriers was elaborated on the basis of the results of the experiments in which different methods of solid dispersion preparation (melting, solvent method, different cooling), different concentrations of drug/carriers and molecular weight of Macrogol were tested. The best solid dispersion consisted of: ketoprofen-Macrogol 6000-KLH(T) (1+8. 9+0.1) was chosen to formulate the pellets on the basis of the pharmaceutical availability of ketoprofen from solid dispersion and the physical chemical studies: thermomicroscopic, DSC and X-ray diffraction. The pellets were prepared by the extrusion and spheronization method. The mechanical properties of the pellets as well as ketoprofen released from pellets containing solid dispersion, in comparison with physical mixtures and the drug alone, were evaluated. The increase in the amount of released ketoprofen from solid dispersion pellets was 3.8-times greater than from the pellets containing the drug alone. The stability of solid dispersion pellets was satisfactory. PMID- 11058807 TI - Immobilization of antibodies on alginate-chitosan beads. AB - An anti-hapten IgG was covalently immobilized on glutaraldehyde-activated alginate-chitosan gel beads. The antibody immobilization efficiency was influenced by glutaraldehyde-bead reaction time, IgG concentration and pH. In addition, immobilization conditions such as glutaraldehyde and antibody concentrations influenced antibody hapten binding affinity. The immobilized IgG on the beads was stable and no reduction in the percent binding to hapten was noticed following 25 days of storage. It was concluded that antibodies could be successfully immobilized on alginate-chitosan gel beads. Such a system can be applied for the development of immunoaffinity purification and immunoassays. PMID- 11058808 TI - Physicochemical studies of lidocaine-menthol binary systems for enhanced membrane transport. AB - The melting properties of lidocaine and l-menthol binary systems were studied using differential scanning calorimetry (DSC). A eutectic mixture was obtained for the lidocaine:menthol ratio of 30:70 (w:w) with a eutectic point of 26 degrees C. The binary melt systems formed within a range of 30:70-50:50 (w:w) remained as homogeneous oils at ambient temperature. The solubilities of pure lidocaine and lidocaine from the binary melt systems were determined with and without propylene glycol in pH 8.0 phosphate buffer. Lidocaine from the melt systems was less soluble in the buffers due to the partition of lidocaine between the oil and aqueous phases. The addition of propylene glycol to the buffer significantly increased both the solubility and heat of solubilization of lidocaine. The permeation rates of lidocaine from the binary melt systems across shed snake-skin were concentration dependent and significantly higher than those from the reference solutions. PMID- 11058809 TI - Effect of nucleoside analogues and oligonucleotides on hydrolysis of liposomal phospholipids. AB - The hydrolysis of the bilayer forming phospholipids resulting first of all in lysophospholipids and fatty acids is one limiting factor determining the shelf life of liposomes. In several studies the influence of pH, buffer, lipid composition and other parameters on the hydrolysis of phospholipids have been demonstrated, but the influence of drugs has not yet been investigated systematically. In this study the influence of nucleoside analogues, especially 2', 2'-difluoro 2'-deoxycytidine (gemcitabine, dFdC) on the degradation of phospholipids was elucidated in more detail. It could be demonstrated that the interaction of dFdC with phospholipid bilayers promotes the hydrolysis of phospholipids in a concentration-dependent manner. Obviously two parts of the molecule, the amino group bound to the pyrimidine moiety and the 2', 2'-difluoro 2'-deoxyribose, seem to be responsible for the forced phospholipid hydrolysis. The dFdC-induced hydrolysis of phospholipids was influenced by pH, buffer, lipid composition and different anions. Optimization of the above parameters resulted in prolonged shelf-life of dFdC liposome dispersions, which is an important prerequisite for clinical practice. PMID- 11058810 TI - Robustness testing, using experimental design, of a flow-through dissolution method for a product where the actives have markedly differing solubility properties. AB - The use of experimental design for the robustness testing of a flow-through dissolution method (Ph Eur/USP Apparatus 4) for atovaquone, one of the drug substances in a dual-active anti-malarial tablet formulation, Malarone tablets, is described. This procedure was developed to overcome the suppression of the atovaquone solubility, caused by the presence of the co-drug proguanil hydrochloride and potential imprecision due to the poor solubility of the coating material in the basic dissolution media employed. For this testing a quarter fractional two-level factorial design was applied, assessing six factors in sixteen experiments, with a further six centre points to assess natural experimental variation. Results demonstrate that the method is robust to small changes in all the main factors evaluated at sample times of 30 min or greater. At 15 min, variations in the concentration of sodium hydroxide in the dissolution media, peristaltic pump speed and flow rate were assessed as statistically significant. This observation is a result of the initial steepness of the dissolution release curve and hence these factors are now controlled routinely in the method. Release of this poorly soluble drug is limited at the 45 min time point (Q=75%) according to pharmacopoeial guidelines. The approach may be applied for other dissolution procedures. PMID- 11058811 TI - Quantitative analysis of polymorphs in binary and multi-component powder mixtures by near-infrared reflectance spectroscopy. AB - Near-infrared reflectance spectroscopy was employed to quantify polymorphs in binary and multi-component powder mixtures. Sulfamethoxazole (SMZ) forms I and II were used as model polymorphs for this study. The instrument reproducibility, method error, precision, and limits of detection and quantification of the method were assessed. Physical mixtures of the polymorph pair were made by weight, ranging from 0 to 100% SMZ form I in II. Near-infrared spectra of the powder samples contained in glass vials were obtained over the wavelength region of 1100 2500 nm. A calibration plot was constructed by plotting SMZ form I weight percent against a ratio of second derivative values of log(1/R') (where R' is the relative reflectance) versus wavelength. The coefficients of determination, R(2), were generally greater than 0.9997 and standard errors were low for all the systems. Instrument error was assessed by analyzing a sample 10 times without perturbation. Method error was assessed in the same manner except the sample was re-mixed between analyses. A precision study was conducted by analyzing aliquots from a larger homogeneous sample. Limits of detection (LOD) and quantification (LOQ) were determined from the standard deviation of the response of the blank samples (100% SMZ form II, undiluted or diluted with 60% lactose). These limits were subsequently validated with independent samples. The results show that polymorphs can be quantified in binary and multi-component mixtures in the 2% polymorph composition range. These studies indicate that NIRS is a precise and accurate quantitative tool for determination of polymorphs in the solid-state, is comparable to other characterization techniques, and is more convenient to use than many other methods. PMID- 11058812 TI - Effect of particle size, air flow and inhaler device on the aerosolisation of disodium cromoglycate powders. AB - Recently, the dispersion of mannitol powders has demonstrated the importance of particle size, air flow and inhaler device (Chew and Chan, 1999). The aim of the present study is to extend our investigation to a different compound, disodium cromoglycate (DSCG) powders. Solid state characteristics of the powders were assessed by particle sizing, scanning electron microscopy, X-ray powder diffraction, moisture content, particle density determination and freeze fracture. The aerosol behaviour of the powders was studied by dispersion using Rotahaler(R) and Dinkihaler(R), connected to a four-stage liquid impinger operating at 30-120 l/min. Three amorphous powders with a mass median diameter (MMD) of 2.3, 3.7, 5.2 microm and a similar polydispersity were prepared. The particles were nearly spherical with a particle density of 1.6 g/cm(3) and moisture content of 6.6 wt.%. Using Rotahaler(R), the maximum fine particle fraction (FPF(max)) for all three powders was only 15 wt.%, attained at the highest flow of 120 l/min. Using Dinkihaler(R), the FPF(max) was two to four times higher, being 36 and 29 wt.% for the 2.3 and 3.7 microm powder, respectively, at 60 l/min; and 18 wt.% for the 5.2 microm powder at 120 l/min. Hence, the study shows that the FPF in the DSCG powder aerosols was determined by the interaction of the particle size, air flow and inhaler design. The attribution of the amorphous nature and the different physico-chemical properties of the powder may explain the incomplete and low dispersibility of DSCG. PMID- 11058813 TI - Microcrystalline cellulose from soybean husk: effects of solvent treatments on its properties as acetylsalicylic acid carrier. AB - Microcrystalline cellulose (MCC) is a very important product in pharmaceutic, food, cosmetic and other industries. In this work, MCC was prepared from soybean husk, produced in large quantities in soybean oil processing industries. It was characterized through various techniques (scanning electron microscopy (SEM), infrared spectroscopy (FTIR), thermogravimetry analysis (TGA) and differential scanning calorimetry (DSC)) and compared with a commercial MCC. The results obtained show that the prepared sample has similar crystallinity and lower particle size than the commercial MCC. Both MCC samples were treated with organic solvents (chloroform, acetone, ethanol and ethyl ether), for structural modifications to be introduced, and used as acetylsalicylic acid (ASA) carrier. Pretreated MCC and MCC/ASA 1:1 mixtures were analyzed through FTIR and thermal analysis. The drug release was evaluated in buffer solution of pH 4.5 and in pure water, at 37 degrees C. The MCC pretreated with different solvents show different thermal properties and ASA release rates, each MCC showing a particular behavior. PMID- 11058815 TI - Erratum to 'Proniosomes: A novel drug carrier Preparation'. PMID- 11058814 TI - High-efficiency gene transfection of macrophages by lipoplexes. AB - Macrophage transfection studies are crucial for understanding gene regulation and expression. However, gene transfection in macrophages is difficult. We have shown here that macrophages are more resistant to gene transfection compared with other cell types. To further develop an efficient gene delivery system for macrophages, we evaluated various liposomal and non-liposomal agents including LipofectAMINE(R), Lipofectin(R), DOTAP, DEAE-dextran, and the DNA condensing agent protamine sulfate for their ability to promote gene transfection. CMV luciferase was used as a reporter plasmid. Macrophage transfection was maximal at the DNA:LipofectAMINE:protamine ratio of 1:12:1 microg/ml. The LipofectAMINE formulation showed a 10-12-fold increase in transfection efficiency over DOTAP and a 4-5-fold increase over Lipofectin. This transfection method showed minimal toxicity at the concentrations tested and was at least 20-25-fold superior to the most frequently used DEAE-dextran method for macrophage transfection. PMID- 11058817 TI - Episodic memory in single cells. PMID- 11058816 TI - Emotional Stroop effect? PMID- 11058818 TI - Object knowledge in infancy: current controversies and approaches. AB - Studies relying on looking-time measures have found evidence of a far more precocious understanding of hidden objects than Piaget originally described. However, there is now a heated controversy surrounding the results from looking time studies - do they constitute any evidence of a conceptual or explicit understanding of objects? Moreover, even within the looking-time paradigm, young infants show rapid changes in their understanding of what constitutes a legitimate occlusion event, and in their ability to use feature information to individuate or keep track of the number of hidden objects. The picture that emerges from these studies is that young infants have a limited and sometimes fragmented understanding of hidden objects. We suggest that computational modelling could help provide a coherent account of the emergence of object directed behaviours in infancy, although the fit between current models and existing data remains poor. PMID- 11058819 TI - The episodic buffer: a new component of working memory? AB - In 1974, Baddeley and Hitch proposed a three-component model of working memory. Over the years, this has been successful in giving an integrated account not only of data from normal adults, but also neuropsychological, developmental and neuroimaging data. There are, however, a number of phenomena that are not readily captured by the original model. These are outlined here and a fourth component to the model, the episodic buffer, is proposed. It comprises a limited capacity system that provides temporary storage of information held in a multimodal code, which is capable of binding information from the subsidiary systems, and from long-term memory, into a unitary episodic representation. Conscious awareness is assumed to be the principal mode of retrieval from the buffer. The revised model differs from the old principally in focussing attention on the processes of integrating information, rather than on the isolation of the subsystems. In doing so, it provides a better basis for tackling the more complex aspects of executive control in working memory. PMID- 11058820 TI - Forward modeling allows feedback control for fast reaching movements. AB - Delays in sensorimotor loops have led to the proposal that reaching movements are primarily under pre-programmed control and that sensory feedback loops exert an influence only at the very end of a trajectory. The present review challenges this view. Although behavioral data suggest that a motor plan is assembled prior to the onset of movement, more recent studies have indicated that this initial plan does not unfold unaltered, but is updated continuously by internal feedback loops. These loops rely on a forward model that integrates the sensory inflow and motor outflow to evaluate the consequence of the motor commands sent to a limb, such as the arm. In such a model, the probable position and velocity of an effector can be estimated with negligible delays and even predicted in advance, thus making feedback strategies possible for fast reaching movements. The parietal lobe and cerebellum appear to play a crucial role in this process. The ability of the motor system to estimate the future state of the limb might be an evolutionary substrate for mental operations that require an estimate of sequelae in the immediate future. PMID- 11058821 TI - Event-related potential studies of attention. AB - Over the past 30 years, recordings of event-related potentials (ERPs) from normal individuals have played an increasingly important role in our understanding of the mechanisms of attention. This article reviews some of the recent ERP studies of attention, focusing on studies that isolate the operation of attention in specific cognitive subsystems such as perception, working memory, and response selection. Several conclusions are drawn. First, under some conditions attention modulates the initial feedforward volley of neural activity in intermediate visual processing areas. Second, these early effects can be observed for both the voluntary allocation of attention and for the automatic capture of attention following a peripheral visual transient. Third, these effects are present not only when attention is directed to a location in 2-dimensional space, but also when attention is directed to one of two spatially overlapping surfaces. Fourth, attention does not modulate sensory activity unless sensory systems are overloaded; when sensory systems are not taxed, attention may instead operate to influence memory or response processes. That is, attention operates to mitigate information overload in whichever cognitive subsystems are overloaded by a particular combination of stimuli and task. PMID- 11058822 TI - In vivo subcellular elemental dynamics in liver graft: With special reference to effect of non-selective endothelin receptor antagonist, TAK-044, on the graft injury. AB - Background: No data are available concerning the in vivo subcellular dynamics of elements in liver grafts and the effect of endothelin receptor antagonist, TAK 044, against graft injury. Methods: Liver transplantation was performed in porcine under active veno-venous bypass. The grafts stored in chilled preservation solution were recirculated following reflush with lactated Ringer's solution with or without TAK-044 (10 mg/kg). Cold and warm ischemic times of the grafts were comparable between the two groups. Elements (Na, K, Cl, Ca, P and S) were measured in three fractions of cytoplasm, mitochondria and nucleus by electron probe X-ray microanalysis for the graft biopsy specimens obtained at various time from donor laparotomy to 1 week after liver grafting. Liver functions also were compared between the two groups. Results: In both groups, concentration of each element changed in parallel among the three subcellular fractions and their changes were less marked in the nucleus. In the control group, there were significant increases in cytoplasmic Na and Cl after portal reperfusion and in cytoplasmic and mitochondrial Ca after hepatic artery reperfusion. These were accompanied by K and mitochondrial S decreases without a statistical significance. In the TAK group, such postreperfusion elemental alterations were significantly suppressed and early deterioration of the liver functions was alleviated, as compared with the control group. Conclusion: A supplemental use of TAK-044 in a rinse solution before reflush contributed to stability of subcellular elements after reperfusion and better preservation of early graft function. PMID- 11058823 TI - Improvement of fuel metabolism by nocturnal energy supplementation in patients with liver cirrhosis. AB - Aims: patients with liver cirrhosis exhibit abnormal fuel metabolism, including increased fat and decreased glucose oxidation. Such altered energy metabolism is similar to that observed after starvation and could lead to malnutrition. We therefore studied whether nocturnal energy supplementation might improve the fuel metabolism in cirrhotic patients. Methods: 12 cirrhotic patients and 14 healthy controls participated in this study. Subjects in the two groups ate isonitrogenous (1.2 g/kg/day) and isocaloric (35 kcal/day) diets for 1 week before and during the study. On day 1 of the study, indirect calorimetry was carried out in the morning after an overnight fast. The next morning, the same measurement was performed after the patients took a liquid nutrient (Ensure Liquid(R), 250 kcal) at 23:00 on day 1. Respiratory quotient (RQ), resting energy expenditure (REE), and substrate oxidation rates of glucose (% CHO), fat (% FAT) and protein were estimated from measured VO(2), VCO(2) and urinary nitrogen. Results: Significant decreases in RQ, and % CHO and a significant increase in % FAT were observed at baseline in cirrhotic patients as compared with controls. After the nocturnal energy supplementation, RQ, % CHO and % FAT in cirrhotic patients were significantly recovered, ending at levels close to normal. Conclusions: These results suggest that nocturnal energy supplementation could be useful to correct abnormal fuel metabolism and to prevent malnutrition in cirrhosis. PMID- 11058824 TI - The role of hepatitis C virus infection in glomerulopathy. AB - The characteristics and prevalence of hepatitis C virus (HCV)-associated glomerulopathy remain to be determined. To analyze the relationship between HCV infection and glomerulopathy, we enrolled three groups of individuals or patients. The first group consisted of 7776 individuals who were seen for routine checkups. The second group consisted of 86 patients with chronic hepatitis C, 40 patients with chronic hepatitis B, and 51 patients with non-viral liver diseases. The third group consisted of nine patients with HCV association glomerulopathy who underwent renal biopsy. Of the 7776 individuals undergoing medical checkups, 142 (1.8%) were positive for HCV antibody. The positive rate of proteinuria was significantly higher (P<0.030) in individuals with HCV antibody (2.1%) than in those without the antibody (0.6%). Abnormal levels of serum creatinine (5.8 vs. 0%, P=0.025) and complications of cryoglobulinemia (45 vs. 5%, P<0.001) were significantly more common in the 86 patients with chronic hepatitis C (5.8%) than in the 91 patients with other liver diseases. All patients with abnormal levels of serum creatinine had concomitant cryoglobulinemia. Of the nine patients with histologically proven HCV-associated glomerulopathy, four had cryoglobulinemia (all were type II). Elevations of serum creatinine level (4/4 vs. 0/5, P=0.048) and a glomerular legion of membranoproliferative glomerulonephritis (3/4 vs. 0/5, P=0.048), a severe type of glomerulonephritis, were more common in the four patients with cryoglobulinemia than in the remaining five patients. In conclusion, HCV infection was found to be significantly associated with glomerulopathy. In addition, the presence of cryoglobulinemia, which usually accompanies membranoproliferative glomerulonephritis, was found to be an indicator of renal insufficiency in patients with HCV-associated glomerulopathy. PMID- 11058825 TI - Late administration of dimethyl sulfoxide minimizes the hepatic microvascular inflammatory response to chloroform in rats. AB - Dimethyl sulfoxide (DMSO) protects against liver injury elicited by chloroform (CHCl(3)) in rats even when given 24 h after the toxicant. Aminobenzotriazole (ABT) is a cytochrome P-450 inhibitor as is DMSO. The present study was conducted to examine the effect of DMSO or ABT on the hepatic microvascular inflammatory response to CHCl(3) using in vivo microscopy. Rats received 0.75 ml/kg CHCl(3) by oral administration followed 24 h later by either 2.0 ml/kg DMSO, 30 mg/kg ABT, or saline injected intraperitoneally. Untreated animals served as controls. At 28 h after CHCl(3) administration, the number of leukocytes adhering to the sinusoidal wall was significantly increased associated with a reduction in the numbers of perfused sinusoids. DMSO, but not ABT, caused a restoration of perfused sinusoids to 85% of normal. At 32 h after, leukocyte adhesion and Kupffer cell phagocytic activity were significantly increased. The numbers of perfused sinusoids were decreased by 32% in periportal regions and 50% in centrilobular regions. Sinusoidal perfusion and Kupffer cell activity were modified significantly by either DMSO or ABT, while leukocyte adhesion was reduced by DMSO alone. The results suggest that attenuation of the hepatic microvascular response to CHCl(3) is a potential mechanism of post-treatment with DMSO. PMID- 11058826 TI - In vitro cytokine production of peripheral blood mononuclear cells in response to HCV core antigen stimulation during interferon-beta treatment and its relevance to sCD8 and sCD30. AB - We investigated the responses of peripheral blood mononuclear cells (PBMCs) to hepatitis C virus core protein in ten patients with chronic hepatitis C during interferon (IFN)-beta treatment to determine if the modulation of the immune reaction to hepatitis C virus by IFN treatment is associated with the viral clearance. Interleukin-2, interleukin-4, interleukin-10, and interferon-gamma in the supernatant of the patients' PBMC co-cultured with the HCV core antigen presenting autologous PBMCs were measured by ELISA. Serum levels of soluble CD (sCD) 8 and sCD30 in these patients were also measured by ELISA. The production of interleukin-2 and interferon-gamma by PBMCs of sustained responders (SRs) increased after IFN-treatment, although it did not reach a significant level. Interleukin-10 was detected only in non-responders (NRs) at 0 and 4 weeks after the start of IFN treatment. Serum sCD8 level increased significantly in SR with IFN treatment. A close correlation between the serum sCD8 levels and interferon gamma levels in the supernatant at week 8 was observed in SR. These results suggest that IFN treatment potentiates the cellular immune reaction against HCV core protein more efficiently in SR than in NR. PMID- 11058827 TI - Second generation amplicor-HCV monitor assay: clinical features and predictors of the response to interferon. AB - The aims of this study were to compare the amplicor-HCV monitor assay versions 1.0 and 2.0, and to investigate the clinical usefulness of this assay in patients with chronic hepatitis C. We retrospectively analyzed 154 patients, and 133 of these patients received interferon therapy. Sixty-nine patients were complete responders (CR), and 64 were non-responders. Serum HCV RNA levels of version 1.0 and version 2.0 and HCV genotypes were determined in all patients. There was a good correlation between versions 1.0 and 2.0 in both genotype 1b and 2a, 2b (r=0.907 and 0.726, respectively). In genotype 1b, the mean HCV RNA level obtained by version 1.0 was 384+/-547 kcopies/ml and that obtained by version 2.0 was 488+/-825 kI.U./ml. In genotype 2a/2b, the mean level obtained by version 1.0 was 170+/-369 kcopies/ml and that obtained by version 2.0 was 340+/-402 kI.U./ml. Discriminant analysis revealed that the discriminating points of IFN response were 168 kcopies/ml (genotype 1b, version 1.0), 106 kcopies/ml (genotype 2a and 2b, version 1.0), 102 kI.U./ml (genotype 1b, version 2.0), and 277 kI.U./ml (genotype 2a and 2b, version 2.0). When the patients were stratified according to the discriminating points, the CR rate below the discriminating points were 73.8 and 86.2% in versions 1.0 and 2.0, respectively, in genotype 1b, and the rates were 73.2 and 82.3% in genotype 2a/2b. In addition, receiver-operating characteristic analysis revealed that version 2.0 had significantly better discriminative ability in patients with genotype 1b. We conclude that the second version of the amplicor-HCV monitor assay measures HCV RNA levels with the same precision as version 1.0 and is more useful for the prediction of interferon response than version 1.0. PMID- 11058828 TI - Evaluation of balloon-occluded hepatic venography with carbon dioxide for portography and correlation between retrograde portogram and liver function in patients with liver cirrhosis. AB - Background/Aim: Hepatic venography with a positive-contrast medium has been reported as a method for evaluating liver disease. However, the contrast medium used in this method provides insufficient portal vein observation and may cause severe liver injuries. Carbon dioxide (CO(2)), a negative-contrast medium, may be able to depict the portal vein system with minimal hepatic toxicity. The aim of this study was to evaluate the usefulness and side-effects of balloon-occluded hepatic venography with CO(2) (CO(2) venography) and to evaluate the correlation between retrograde portogram and liver function in patients with cirrhosis. Subjects and methods: The subjects consisted of 23 biopsy-proven cirrhotic patients (male:female, 16:7; age, 58+/-12 years, range 34-80). The causes of cirrhosis were alcohol intake in ten, HCV infection in ten, HBV infection in one, primary biliary cirrhosis in one and Budd-Chiari syndrome in one. Of these patients, six were complicated with hepatocellular carcinoma (HCC). CO(2) venography was performed with an occlusion balloon catheter, and 25 ml of CO(2) was infused. CO(2) portograms were scored as follows: 0, no visualization of portal veins; 1, visualization of peripheral portal branches; 2, unilateral first portal branch; 3, bilateral first portal branches; 4, main portal vein; 5, left gastric vein, superior mesenteric vein and splenic vein. Hepatic venous pressure gradient (HVPG), cardiac functions, biochemical analysis, blood gas analysis and oxygen (O(2)) saturation monitoring were measured simultaneously. Arterio portography was also performed. To evaluate the usefulness of CO(2) venography in patients with HCC accompanied by portal vein tumor thrombus (PVTT), three patients were also examined. Results: No significant changes in ALT, AST, O(2) saturation or blood gas data were observed after CO(2) venography. A statistically significant positive correlation was observed between CO(2) portogram scores and Child-Pugh scores (r=0.68, P=0.003). The correlations between CO(2) portogram scores and HVPG, and the forms of gastroesophageal varices in patients without PVTT and major shunts were not significant. The CO(2) portogram score was significantly higher in patients with alcoholic liver cirrhosis than in those with HCV-positive cirrhosis (P=0.04). Cavernous transformation and peripheral portal branches were demonstrated in patients with HCC accompanied by PVTT. These findings could not be observed by arterio portography. Conclusion: CO(2) venography to obtain retrograde portogram is a safe and useful method for evaluating the portal vein system and liver function in patients with liver cirrhosis. PMID- 11058829 TI - Distinction between chronic hepatitis and liver cirrhosis in patients with hepatitis C virus infection. Practical discriminant function using common laboratory data. AB - In order to distinguish patients with cirrhosis from those with chronic hepatitis, multivariate discriminant analysis was performed using common laboratory data. A total of 205 consecutive patients were diagnosed by peritoneoscopy and biopsy as having chronic liver disease caused by hepatitis C virus (HCV), 168 with chronic hepatitis and 37 with cirrhosis. Twenty variables and their natural logarithmic transformation were employed in the multivariate analysis. After stepwise variable selection, the following function was finally obtained to discriminate the disease severity, z=0.120xgamma-globulin (%)+0.423xln (hyaluronate) (ug l(-1))-0.059xplatelet (x10(4) counts per mm(3)) 0.364xsex (male, 1; female, 2)-3.953. Since the function contained an expression of logarithm and was slightly troublesome to apply, we prepared another discriminant function composed of usual figures without logarithmic transformation, z=0.124x(gamma-globulin (%))+0.001x(hyaluronate) (ug l(-1)) 0.075x(platelet (x10(4) counts per mm(3)))-0.413xgender (male, 1; female, 2) 2.005. When a positive result is calculated in the latter equation, the diagnosis of the liver disease indicates cirrhosis, and negative result chronic hepatitis. Accuracy of the two discriminant functions was 90.3 and 91.2%, respectively. A concise linear discriminant function could successfully differentiate liver cirrhosis from chronic hepatitis with an accuracy of 91.2%. PMID- 11058830 TI - Amino acid mutations in the interferon sensitivity determining region and serum virus load in hepatitis C virus carriers with long-term normal ALT levels. AB - The amino acid mutations in a part of the non-structural region 5A (NS5A) of the hepatitis C virus (HCV) genome, called the interferon sensitivity determining region (ISDR), can affect the response to interferon (IFN) treatment. We analyzed the serial changes of the amino acid substitutions in the ISDR during the natural course of patients with sustained long-term normal alanine aminotransferase (ALT) levels in relation to the changes in virus load, and assessed the clinical significance of ISDR in the natural course and IFN treatment. The subjects were nine patients infected with HCV (genotype 1b) who had been examined for serum ALT levels every month for more than 1 year and had well-sustained normal levels. The amino acid sequence of the ISDR was determined by the direct sequencing method, and the number of amino acid mutations was evaluated in comparison with the prototype (HCV-J). Quantitation of serum HCV RNA levels was conducted by the Amplicor-monitor method (Nihon Roche). On the initial analysis of the ISDR, six patients were determined to have no mutations, and three patients had one or two mutations. However, an increased number in amino acid mutations compared with the wild type during the follow-up period was confirmed in only one patient, and that increase was limited to within two amino acids. Virus load changed regardless of the changes in amino acid substitutions in the ISDR. The ISDR was therefore inferred to be a stable region unrelated to the virus load in patients with well sustained normal ALT levels. Additional changes of amino acid sequence in this region were not a sensitive marker for determining whether IFN treatment is indicated. PMID- 11058831 TI - Mutations in the non-structural protein 5A gene in patients with chronic hepatitis C virus 1b infection during repeated interferon treatment. AB - It has been previously reported that the non-structural region 5A (NS5A) of the hepatitis C virus (HCV) includes an interferon sensitivity determining region (ISDR) and that amino acid substitutions in this region are closely associated with the response to interferon (IFN) treatment. We assessed the clinical significance of serial changes of amino acid sequences in the ISDR during repeated IFN treatment in patients with chronic hepatitis C (genotype 1b), related to serum HCV RNA load. During treatment, additional amino acid substitutions in the ISDR were observed in four of eight patients (50% 2/5 of complete responders (CR); 2/3 of non-responders (NR). However, comparing these amino acid substitutions to wild-type ISDR, the number of amino acid mutations was limited to only one amino acid identified in two CRs. The virus load changed regardless of the amino acid substitutions in the ISDR during treatment, and the wild-type and intermediate type (with less than three amino acid substitutions) showed wide variations in virus load. These data indicate that amino acid mutations in the ISDR, which indicate the switch to mutant-type do not occur easily even during repeated IFN treatment, and the additional amino acid substitutions in the ISDR are not a sensitive marker during repeated IFN treatment. In cases where virus load is used as a marker of response to repeated UN treatment, serial examinations are necessary to determine the precise virus load levels. PMID- 11058832 TI - Beneficial effect of Cyclosporin A on acute hepatic injury induced by galactosamine and lipopolysaccharide in rats. AB - Cyclosporin A (CsA), a potent immunosuppressive agent, is used clinically to prevent allograft rejection after organ transplantation. Although recent experimental studies show that CsA prevents severe hepatic injury associated with tumor necrosis factor-alpha (TNF-alpha), the efficacy of CsA has not been confirmed. In the present study, we evaluated whether CsA protects against the severe hepatic injury induced by simultaneous administration of D-galactosamine (GaIN, 200 mg/kg) and lipopolysaccharide (LPS, 10 ug/kg) into the portal vein. The method of CsA (10 mg/kg) was divided into four groups; i.e. preinjection (24 and 2 h before administration of GaIN and LPS), concomitant single-injection, multiple-interval injection (0, 3, 6, 9 and 15 h after administration of GaIN and LPS) and no treatment. In the group receiving multiple-interval injections of CsA, liver function parameters (total bilirubin, asparate aminotransferase and alanine aminotransferase levels in sera) were significantly improved and the development of massive hepatic necrosis was significantly prevented, but no improvement of liver function parameters or histological changes was shown either the pre- or concomitant single-injection groups. The serum levels of TNF-alpha and interferon-gamma showed no differences between the no treatment and multiple interval injection groups. However, the serum level of interleukin 8 and neutrophil infiltration into the liver tissue after 24 h GaIN and LPS administration were significantly lower than those of the control group. The apoptotic index of liver tissue using the TUNEL method was not significantly different. Our data suggest that CsA protects against acute hepatic injury, if it is administered at the appropriate time during the course of hepatic injury. PMID- 11058833 TI - A case of erythropoietic protoporphyria with liver cirrhosis suggesting a therapeutic value of supplementation with alpha-tocopherol. AB - Erythropoietic protoporphyria (EPP) was diagnosed in a 27-year-old man based on a typical clinical history, and a marked increase in erythrocyte and fecal protoporphyrin concentrations. Although liver disease is not a common feature in EPP, he had slight liver dysfunction. A percutaneous liver biopsy was performed and it showed minimal hepatocellular damage and many reddish brown pigment deposits in hepatocytes, Kupffer cells, portal histiocytes, bile canaliculi and small biliary radicles. Electron microscopic findings confirmed that these deposits were composed of protoporphyrin crystals. Liver biochemistry remained well for >2 years, but deteriorated rapidly and second liver biopsy obtained 28 months after the first biopsy revealed the development of liver cirrhosis. We treated the patient with intravenous administration of dl-a-tocopherol acetate (vitamin E; 500 IU/body/day). Following the administration of vitamin E, the concentration of protoporphyrin in erythrocytes decreased significantly and the liver function tests were improved. Sixteen weeks later he showed the full clinical and biochemical recovery suggesting that vitamin E supplementation might be useful in treating patients with EPP who developed liver damage. PMID- 11058834 TI - Randomized controlled trial of twice-a-day administration of natural interferon beta for chronic hepatitis C. AB - We conducted a randomized controlled trial to assess the efficacy of twice-a-day administration of natural interferon beta (IFNbeta) as an induction of IFN therapy for chronic hepatitis C. Seventy-one patients with chronic hepatitis C were enrolled into the trial and randomly assigned into three treatment groups. Six million units (MU) of IFNbeta were administered once-a-day for the first 4 weeks, and then thrice weekly for 12 weeks in 20 patients (once-a-day group). Three milion units of IFNbeta were administered twice-a-day for the first 2 weeks, 6 MU once-a-day for the next 2 weeks, and then thrice weekly for 12 weeks in 23 patients (twice-a-day+beta group), or 6 MU of lymphoblastoid IFNalpha were administered thrice weekly for the last 12 weeks instead of IFNbeta in 28 patients (twice-a-day+alpha group). Four patients in once-a-day group (20%), 9 in twice-a-day+beta group (39%), and 12 in twice-a-day+alpha group (43%) obtained sustained response. Sustained response rate in twice-a-day groups was higher than in once-a-day group, although there was no statistical significance. The present study suggested the possible superiority of twice-a-day administration of IFNbeta as an induction therapy to once-a-day administration, but further studies are needed to confirm this regimen. PMID- 11058835 TI - Quasi-dynamical electron diffraction - a kinematic type of expression for the dynamical diffracted-beam amplitudes AB - It is shown that to a good approximation the dynamical diffracted electron-beam amplitudes may be expressed in a form that is identical to that of the kinematic theory of electron diffraction. The validity of this approximate form of the dynamical electron diffraction is illustrated for thin films of GaAs and Au crystals, and its implications in electron crystallography for structural determination and refinement are discussed. PMID- 11058836 TI - Debye-waller factors of compounds with the caesium chloride structure AB - The lattice dynamics of five compounds with the caesium chloride structure have been investigated using shell models. Debye-Waller factors for these compounds are calculated over the temperature range from 1 to 1000 K and the results are presented analytically in a polynomial form. When experimental results are available, the calculated results are compared to the experimentally measured Debye-Waller factors and typically the discrepancies between these factors are less than 10%. PMID- 11058837 TI - A hybrid minimal principle for the crystallographic phase problem. AB - Simulated annealing is used to solve the X-ray phase problem formulated as a minimization problem. The cost function consists of two parts, one represents the discrepancy between measured and calculated intensities while the other monitors the probability distribution of the triplets. From a random real-space structure at the start, the atoms are moved one by one to gradually reduce the cost function until the best structure emerges. Trial calculations for structures including hexadecaisoleucinomycin (HEXIL) are presented. Comparison of this method with other related methods is made. PMID- 11058838 TI - Low-resolution direct phase determination in protein electron crystallography - breaking globular constraints. AB - Although the assumption of an overall globular scattering entity can be useful for determining crystallographic phases for a protein at low resolution, there is a point where this pseudoatomic model must be abandoned for further phase refinement. Using 6 A resolution electron diffraction data from aquaporin (AQP CHIP) as an example, phases of the 16 most intense reflections from a previous direct solution (Dorset & Jap (1998). Acta Cryst. D54, 615-621) were modified with a Hadamard error-correcting code to produce potential maps very similar to the ones obtained using phases from the Fourier transform of averaged electron micrographs. The choice of the optimal phase set was made via the cross correlation of experimental with anticipated density histograms using the autocorrelation function of the latter histogram as the desired endpoint. PMID- 11058839 TI - A model for diffraction from MCM-41 materials AB - A model involving only a small number of parameters provides a convenient way of interpreting diffraction patterns from MCM-41 materials. Each parameter of the model has a clear physical meaning, and this approach is clearly superior to extracting pore structure information by fitting Gaussians to an observed diffraction pattern. PMID- 11058840 TI - The statistical kinematical theory of X-ray diffraction as applied to reciprocal space mapping AB - The statistical kinematical X-ray diffraction theory is developed to describe reciprocal-space maps (RSMs) from deformed crystals with defects of the structure. The general solutions for coherent and diffuse components of the scattered intensity in reciprocal space are derived. As an example, the explicit expressions for intensity distributions in the case of spherical defects and of a mosaic crystal were obtained. The theory takes into account the instrumental function of the triple-crystal diffractometer and can therefore be used for experimental data analysis. PMID- 11058841 TI - The crystallite size-disorder relationship based on the spiral paracrystal. AB - A simple physical model of the relationship between crystallite size and disorder for paracrystalline materials is presented, based on the spiral paracrystal and distortion of the lattice cells. Simulations show that the model leads to relationships similar to the alpha* rule. Average crystallite sizes predicted by the model are in agreement with experimental data and also allow crystallite-size distributions to be proposed. The model provides a more satisfactory and complete explanation of this relationship than do current descriptions. PMID- 11058842 TI - Resolution of the phase-ambiguity problem in the centrosymmetric P [1] space group by Monte Carlo methods. AB - Simulated-annealing methods have been used to resolve the phase-ambiguity problem in the centrosymmetric P?bar?1? space group. First, an energy function based on the Sayre equation is introduced and a formal comparison with classical spin systems is drawn. The energy landscape is studied in detail and the validity of several energy criteria thoroughly tested. Classical Monte Carlo methods proved to be successful using a multistart optimization of the Sayre score, along with the additional monitoring of other energetic criteria. These involved the Terwilliger map quality index in reciprocal space in the absence of envelope information, and an envelope score if the shape of the molecule is known. The inherent phase-ambiguity problem of the P?bar?1? space group was therefore technically solved by Monte Carlo methods. The method should also work to resolve phase ambiguity in the SIR method of protein crystallography. PMID- 11058843 TI - General formalism for phase combination and phase refinement: a statistical thermodynamics approach in reciprocal space. AB - The mean-field optimization methodology has been used to recast in a single formalism the problem of phase optimization using an arbitrary energy function in the presence of an experimentally determined phase probability distribution function. It results naturally in the generalization of the notions of figure of merit and centroid phase where the weight of the energy refinement is controlled by an effective temperature in a self-consistent manner. In the limit of high temperature, the formalism reduces of course to the Blow & Crick [Acta Cryst. (1959), 12, 794-802] classical treatment. If a model is available, Sim's [Acta Cryst. (1960), 13, 511-512] weighting scheme for a combined map appears as the first step of a refinement to be conducted until self-consistency is achieved. Assuming that MIR phases exist and that they agree reasonably well with the phases of the model, a first-order expansion gives an estimate of the changes of weights and phases to be performed for the Fourier synthesis. This provides for a new way of doing phase combination that might prove useful in challenging cases of model refinement, e.g. in large macromolecular complexes. Thermodynamic considerations have been used to discuss the best determination of weights in phase refinement; they also suggest that a variational expression of maximum likelihood is best suited as a target for refinement because it is the free energy of the system. The formalism readily allows use of solvent flattening, density averaging and the atomicity criterion to refine phases, and automatically assigns a figure of merit to each reflection. Numerical tests of the method are presented in an attempt to resolve the phase-ambiguity problem of protein crystallography in the centrosymmetric P?1? space group using an energy derived from the Sayre equation. PMID- 11058844 TI - Topological analyses of cuprite, Cu2O, using high-energy synchrotron-radiation data AB - Topological analyses were performed on room-temperature and low-temperature high energy synchrotron-radiation data from cuprite, Cu(2)O, using both single exponential and analytical functions in the multipole model. Differences between the refinements of the data sets turned out to be small and the data were found to be well suited for the analyses and for evaluations of further properties, e. g. the electrostatic potential and the Laplacian. The ionic character of the copper-oxygen bond was impressively underlined by the results. On the 'empty' tetrahedral sites, minima of the charge density were always found. PMID- 11058845 TI - The experimental charge-density approach in the evaluation of intermolecular interactions. Application of a new module of the XD programming package to several solids including a pentapeptide. AB - A new module interfaced to the XD programming package has been used in the evaluation of intermolecular interactions and lattice energies of the crystals of p-nitroaniline, L-asparagine monohydrate and the pentapeptide Boc-Gln-D-Iva-Hyp Ala-Phol (Boc = butoxycarbonyl, Iva = isovaline = ethylalanine, Phol = phenylalaninol). The electrostatic interactions are evaluated with the atom centered distributed multipoles from KRMM (kappa'-restricted multipole model) refinements, using the Buckingham expression for non-overlapping charge densities. Results for p-nitroaniline are compared with Hartree-Fock (HF), density functional (DFT) and Moller-Plesset (MP2) supermolecular calculations and with HF and DFT periodic calculations. The HF and DFT methods fail to predict the stability of the p-nitroaniline crystal but the results of the experimental charge-density approach (ECDA) are in good agreement with both MP2 interaction energies and the experimental lattice energy. ECDA results for L-asparagine monohydrate compare well with those from DFT supermolecular and periodic HF calculations. The disorder of the terminal group in the pentapeptide, which persists at the experimental temperature of 20 K, corresponds to an energy difference of only 0.35 kJ mol(-1), which is too small to be reproduced with current methods. PMID- 11058846 TI - Solving crystal structures from two-wavelength X-ray powder diffraction data - breaking the phase ambiguity in the noncentrosymmetric case AB - Direct methods of breaking phase ambiguities in protein crystallography have been introduced in powder diffraction analysis. This is aiming at ab initio solution of noncentrosymmetric structures using two-wavelength anomalous powder diffraction data. The known structure of the hydrogen bromide salt of leotidine (C(14)H(20)O(2)N(2).HBr) in space group P2(1)2(1)2(1) was used for simulating two wavelength anomalous powder diffraction with the Br atom as anomalous scatterer. X-ray wavelengths are selected at lambda(1) = 0.920 and lambda(2) = 1.500 A. Unique reflections from the diffraction pattern of lambda(2) were able to locate the Br atom accurately. All overlapping diffraction peaks were uniformly partitioned to decompose into single reflections. Structure-factor amplitudes were then extracted. With these and the substructure of Br atoms, unique phases for centric reflections (hk0, h0l and 0kl) and phase doublets for noncentric reflections were obtained. The direct method was used to break the phase ambiguity leading to an interpretable electron-density map, from which five cycles of Fourier iteration yielded the complete structure. PMID- 11058847 TI - On possible extensions of X-ray crystallography through diffraction-pattern oversampling AB - It is known that sampling the diffraction pattern of a finite specimen, at a spacing somewhat finer than the Nyquist spacing (the inverse of the size of the diffracting specimen), corresponds to generating a no-density region surrounding the electron density of the specimen. This no-density region can then be used to retrieve the phase information. In earlier papers [Miao, Sayre & Chapman (1998). J. Opt. Soc. Am. A15, 1662-1669; Sayre, Chapman & Miao (1998). Acta Cryst. A54, 232-239], it was demonstrated, in the case of non-crystalline specimens, that this no-density region could be used to retrieve the phase information; here the same is performed for crystalline and near-crystalline specimens. By employment of an iterative algorithm, the phase information could be recovered from computer generated oversampled diffraction patterns of small specimens that are (a) perfect or imperfect crystals, or (b) have a repeated motif without orientational regularity, or (c) are an unrepeated motif, such as an amorphous glass, a single molecule or a single biological cell. Cases (a) and (b) represent an extension over work recently published [Miao, Charalambous, Kirz & Sayre (1999). Nature (London), 400, 342-344]. Our algorithm requires an approximate envelope for the specimen. It does not require any structural knowledge concerning the specimen and does not require data to atomic resolution (although it can use such data if present). After a few hundred to a few thousand iterations, the correct phase set and image are recovered. The oversampling technique thus greatly extends the specimen range of X-ray crystallography but it imposes a high radiation dose on the specimens compared with the situation in crystallography, in which it is usual for the pattern to be sampled at the (much less fine) Bragg spacing (the inverse of the size of the unit cell). In cases where the specimen is a crystal, there are also possibilities for oversampling relative to Bragg (instead of Nyquist) sampling, thus providing a lesser degree of oversampling and the possibility of lower dosage. Damage of the specimen in consequence of the dose will in many cases seriously affect the quality and resolution of the imaging, but in at least one case [the biological cell in (c) above] the imaging obtainable with the aid of a cryogenic protective technique should surpass any other present method of whole-cell imaging. In addition, with the possible appearance in the future of free electron lasers (>10(12) photons and <200 fs per pulse), it is possible to circumvent the radiation-damage problem by recording diffraction patterns before damage manifests itself. PMID- 11058848 TI - New developments of the TWIN algorithm for phase extension and refinement in disordered supramolecular structures. AB - A new development of the TWIN algorithm is described and used for phase extension/refinement in supramolecular complexes. A small number of phased reflections at low resolution are sufficient for the quasi-automated determination of all atomic coordinates, including disordered atoms. PMID- 11058849 TI - International Union of Crystallography. Report of the Executive Committee for 1999. PMID- 11058850 TI - Prices of IUCr journals PMID- 11058851 TI - A randomized prospective study of radially expanding trocars in laparoscopic surgery. AB - Trocar injury is one of the most serious and potentially preventable complications of laparoscopic surgery. Use of a blunt rather than a cutting trocar could be expected to lessen the likelihood of this injury. Therefore complications related to laparoscopic port design were studied by comparing conventional cutting trocars with radially expanding (blunt) trocars. A multicenter, prospective, randomized clinical trial was conducted in 250 adult patients undergoing elective laparoscopic procedures at tertiary care centers and community hospitals. The patients were randomly assigned to one of two groups: group C, conventional cutting trocars; or group S, radially expanding trocars. Sixteen surgeons performed 244 elective laparoscopic procedures; six patients were removed from the study. One hundred nineteen patients were assigned to group S and 125 to group C. The groups were similar with regard to age, sex, and type of procedure. The following data were collected: intraoperative complications related to the trocars, abdominal wall bleeding, visceral or vascular injury, other complications, fascial closure, procedure time, trocar site assessment at 4 and 24 hours postoperatively, and visual analog pain scores at 4, 8, 12, and 24 hours postoperatively. Fascial defects from 10 mm or larger trocars in group C were closed; the fascial defects in group S were not closed. The trocar sites were checked for incisional hernias at late follow-up. Mean operating time was not different between the two groups (group S, 92 +/- 73 minutes; group C, 100 +/ 74 minutes). There were no episodes of intraoperative cannula site bleeding in group S compared with 16 episodes in 13 patients (P < 0.001) in group C. Postoperative wound complications were fewer in group S (13 vs. 23; P < 0.05). Although the pain scores were generally lower in group S, the differences were not significant. Only 3% of the patients in group S had fascial defects of 10 mm or greater that had to be closed. Within a follow-up period of 6 to 18 months, there have been no incisional hernias in either group. This study shows that radially expanding trocars are safe and effective, and less likely than conventional trocars to result in intraoperative or postoperative complications. The defects created by the radially expanding trocars do not have to be routinely closed. PMID- 11058852 TI - Hepatic resection at a community hospital. AB - Hepatic resection remains the "gold standard" in the primary management of primary and metastatic tumors to the liver. Advanced surgical techniques along with more modern and sophisticated equipment have led to an increasing number of hepatic resections being performed with a concomitant decrease in morbidity and mortality. We followed prospectively 18 consecutive hepatic resections performed over a period of approximately 2.5 years. The setting was a community teaching hospital with a low volume of referrals for hepatic resection. Sixteen (88%) had metastatic disease and two had primary liver disease. There were four trisegmentectomies, four lobectomies, four segmentectomies, and six large wedge re-sections. Average estimated blood loss was 608 ml. Seven patients required transfusions. Complications occurred in five patients (27%). In-hospital mortality was 0%. Our experience suggests that liver resections in a low-volume community hospital can be performed safely provided an experienced surgical team with familiarity of advanced surgical techniques and sophisticated equipment used in hepatic resection is readily available. PMID- 11058853 TI - Quality of life and long-term survival after surgery for chronic pancreatitis. AB - The objective of this study was to evaluate the short-term and long-term outcome as well as quality of life in patients undergoing surgical management of chronic pancreatitis. Between January 1980 and December 1996, a total of 255 patients underwent surgery for chronic pancreatitis at The Johns Hopkins Hospital. The etiology of the disease, indications for surgery, patient characteristics, and long-term survival were analyzed. A visual analog quality-of-life questionnaire containing 23 items graded on a scale of 0 to 10 (0 = worst and 10 = best) was sent to patients postoperatively. Visual analog responses relating to before and after the chronic pancreatitis surgery were compared using a paired t test. During the17-year review period, 263 operations were performed for chronic pancreatitis in 255 patients. The most common presenting symptoms were abdominal pain (88%), weight loss (36%), nausea/vomiting (30%), jaundice (14%), and diarrhea (12%). The cause of the pancreatitis was resumed to be alcohol in 43%, idiopathic in 38%, pancreas divisum in 5%, ampullary abnormality in 4%, and gallstones in 3%. Pancreaticoduodenectomy was the most common procedure in 96 patients (37%), followed by distal pancreatectomy in 67 (25%), Puestow procedure in 52 (19%), sphincteroplasty in 37 (14%), and Duval procedure in five (2%). The overall mortality and morbidity rates were 1.9% and 35%, respectively. Two hundred twenty-seven (89%) of the 255 patients were alive at last follow-up. For the entire cohort of patients, the 5- and 10-year actuarial survivals were 88% and 82%, respectively. One hundred six (47%) of the 227 living patients responded to the visual analog quality-of-life questionnaire. Patients reported improvements in all aspects of the quality-of-life survey including enjoyment out of life, satisfaction with life, pain, number of hospitalizations, feelings of usefulness, and overall health (P < 0.005). In addition to improved quality of life after surgery, narcotic use was decreased (41% vs. 21%, P < 0.01) and alcohol use was decreased (59% vs. 33%, P < 0.001). However, patients often became insulin-dependent diabetics (12% vs. 41%, P < 0.0001) and required pancreatic enzyme supplementation (34% vs. 55%, P < 0.01) after surgical intervention. These data suggest that surgery for patients with chronic pancreatitis can be performed safely with minimal morbidity and excellent long term survival. Moreover, this study evaluates quality of life in a standardized analog fashion, with highly significant improvement reported in all quality-of life measures. We conclude that surgery remains an excellent option for patients with chronic pancreatitis. PMID- 11058854 TI - Two-stage trauma pancreaticoduodenectomy: delay facilitates anastomotic reconstruction. AB - A case of a gunshot wound to the head of the pancreas and superior mesenteric vein requiring pancreaticoduodenectomy is discussed. Managing such an injury is challenging, first because of the ongoing hemorrhage and second because of the technical difficulty in working with a normal pancreas and bile duct. In the case presented herein, enteric reconstruction was performed 72 hours after the initial surgery. A delay in reconstruction resulted in tissue changes that facilitated enteric reconstruction A two-stage pancreaticoduodenectomy may be considered if the surgeon is faced with an unstable patient. PMID- 11058855 TI - Specific pancreatic enzymes activate macrophages to produce tumor necrosis factor alpha: role of nuclear factor kappa B and inhibitory kappa B proteins. AB - The triggering events by which mononuclear cells throughout the body are induced to produce large amounts of cytokines during acute pancreatitis are unclear. However, recent work in our laboratory demonstrated that three specific pancreatic enzymes (elastase, carboxypeptidase A, and lipase) induced dramatic tumor necrosis factor-alpha (TNF-alpha) protein production from macrophages, whereas all others could not. This series of experiments was designed to examine the second messenger system by which this occurs. The rat macrophage cell line NR8383 was incubated for 3 hours with elastase, carboxypeptidase A, lipase, trypsin, or lipopolysaccharide (positive control). Activation of nuclear factor kappa B (NF-kappa B) was demonstrated by electrophoretic mobility shift assay, presence of inhibitory kappa B alpha and beta (I kappa B-alpha and I kappa B beta) by Western blot analysis, and TNF-alpha protein production by enzyme-linked immunosorbent assay. Elastase, carboxypeptidase A, and lipase induced degradation of I kappa B-beta (but not I kappa B-alpha), activation of NF-kappa B, and production of TNF-alpha protein, whereas inhibition of I kappa B with pyrrolidine dithiocarbamate attenuated this response. Trypsin was unable to elicit any of these responses. Macrophages can be induced by specific activated pancreatic enzymes-elastase, carboxypeptidase A, and lipase-to produce TNF-alpha. This process is dependent on I kappa B-beta degradation and NF- kappa B activation, suggesting that these enzymes trigger this second messenger system through specific membrane-bound receptors. PMID- 11058856 TI - Peripheral lymphocyte reduction in severe acute pancreatitis is caused by apoptotic cell death. AB - To investigate impairment of cellular immunity in severe acute pancreatitis, alterations of peripheral lymphocytes in acute pancreatitis were examined. In 48 patients with severe acute pancreatitis, the mean peripheral lymphocyte count on admission was 959 + 105/mm3, and it was significantly decreased in the patients with subsequent infection (623 +/- 90/mm3 ) in comparison to those without infection (1084 +/- 135/mm(3)). According to an analysis of lymphocyte subsets, although both B and T lymphocytes were decreased in peripheral circulation in the patients with infection, it was primarily CD8-positive lymphocytes that decreased in these subsets. Cell cycle analysis of lymphocytes collected from these patients indicated that apoptotic changes occurred after 24 hours' incubation in lymphocytes from patients with severe pancreatitis but not in lymphocytes from healthy control subjects. In a rat model of experimental necrotizing pancreatitis, total peripheral lymphocytes and T lymphocytes were significantly decreased 5 hours after induction of pancreatitis. In severe pancreatitis, peripheral lymphocytes are eliminated from systemic circulation possibly as a result of apoptosis. It has been suggested that impairment of cellular immunity due to peripheral lymphocyte apoptosis is linked to the development of subsequent infectious complications in acute pancreatitis. PMID- 11058857 TI - Somatostatinoma of the ampulla of vater in celiac sprue. AB - The increased incidence of gastrointestinal lymphoma and adenocarcinoma in patients with celiac sprue is well recognized, with 10% to 15% developing a gastrointestinal malignancy. Somatostatinomas are rare neuroendocrine tumors that occur most commonly within the pancreatic head or duodenum. Although fewer than 100 cases have been reported, somatostatinomas are often associated with multiple endocrine neoplasia-1 syndrome and von Recklinghausen's disease. The unusual case of a 43-year-old woman with celiac sprue in which a somatostatinoma involving the ampulla of Vater was identified and resected is presented. To our knowledge, somatostatinomas have not been previously reported in patients with celiac sprue. PMID- 11058858 TI - Anatomic dilatation of the cardia and competence of the lower esophageal sphincter: a clinical and experimental study. AB - Anatomic and clinical data suggest that the gastroesophageal junction or cardia in patients with gastroesophageal reflux disease GERD) may be dilated. We hypothesized that anatomic dilatation of the cardia induces a lower esophageal sphincter dysfunction that may be corrected by narrowing the gastroesophageal junction (i.e., calibration of the cardia). We measured the perimeter of the cardia during surgery in control subjects and patients with GERD and Barrett's esophagus. We then tested our hypothesis in a mechanical model. The model was based on a pig gastroesophageal specimen with perpendicularly placed elastic bands around the cardia simulating the action of the "sling" and "clasp" fibers. "Dilatation" of the cardia was induced by displacing the sling band laterally and decreasing its tension. "Calibration" of the cardia was performed by reapproximation of the sling band toward the esophagus but maintaining the same tension as the dilated model. In the "basal," "dilated," and "calibrated" states, the perimeter of the cardia was noted and rapid mechanized pullback manometry with a water-perfused catheter was performed. The opening pressure was determined, and three-dimensional sphincter pressure images were analyzed. The average cardia perimeter was 6.3 cm in control subjects, 8.9 cm in GERD patients, and 13.8 cm in patients with Barrett's esophagus. The arrangement of the bands in the experimental model generated a manometric high-pressure zone similar to that in the human lower esophageal sphincter. Dilatation of the cardia resulted in a decrease in the resting pressure, length, and vector volume of the high -pressure zone, and reduced the opening pressure. Calibration restored the resting and opening pressure, and normalized the three-dimensional pressure image. In patients with GERD and Barrett's esophagus, the cardia is dilated. Our model supports the hypothesis that lower esophageal sphincter function is compromised by anatomic dilatation of the cardia and can be restored by approximation of the "sling" fibers toward the lesser curvature "clasp" fibers). This provides evidence for a correlation between gastroesophageal sphincter dysfunction in reflux disease and its correction by antireflux surgery. PMID- 11058859 TI - Percutaneous replacement jejunostomy after esophagogastrectomy. AB - A surgically placed jejunostomy tube is a safe and effective means of delivering nutritional support for the postesophagogastrectomy patient. We have previously described a method that permits percutaneous replacement of surgically placed jejunostomy feeding tubes, and now present our results with the use of this technique in 350 consecutive esophagogastrectomy patients. Replacement jejunostomy as required in 17 patients (4.9%). All patients had successful percutaneous jejunostomy replacement. There were no procedural complications or deaths. The timing of feeding tube replacement following esophagogastrectomy was predictive of the indication. Before 16 weeks, the indication for feeding tube replacement was intubation and inability to eat (1 patient) or anorexia with weight loss and dehydration (7 patients). At or after 16 weeks, the indications for feeding tube replacement were all related to symptoms resulting from recurrent carcinoma. We conclude that the technique of percutaneous jejunostomy allows the surgeon tremendous flexibility in the management of the postesophagogastrectomy patient as it preserves the advantages of an adjuvant surgically placed feeding tube over the lifetime of the patient. The technique is safe, and the success rate is excellent. PMID- 11058860 TI - Management of esophageal perforation after pneumatic dilation for achalasia. AB - Current management of esophageal perforation after pneumatic dilation for achalasia is thoracotomy and repair with myotomy. This study aims to assess the outcome of patients managed by laparotomy, and the role of laparoscopic repair. The study was carried out by means of retrospective case review and prospective follow-up with a symptom questionnaire. Results were compared with results in patients undergoing elective Heller myotomy. Over a 20-year period, 445 dilations for achalasia were performed in 371 patients. There were 10 esophageal perforations. Nine patients were referred for surgery and were successfully managed with a transabdominal repair. Laparoscopic repair was attempted in four patients but was successful in only one because of the perforation site. After a mean follow-up of 5.4 years, grade 1 or 2 Visick scores were recorded in all patients. Residual symptoms of dysphagia occurred in 67% in the emergency group and 88% in the elective group. There was an increased incidence of heart-burn compared to elective myotomy. Early operation after perforation provides good results for treatment of achalasia. Mild dysphagia persists and there is an increasing sensation of heartburn. The site of perforation is typically posterolateral, which makes laparoscopic repair difficult. PMID- 11058861 TI - Prevention of mucosal atrophy: role of glutamine and caspases in apoptosis in intestinal epithelial cells. AB - Glutamine starvation induces apoptosis in enterocytes; therefore glutamine is important in the maintenance of gut mucosal homeostasis. However, the molecular mechanisms are unknown. The caspase family of proteases constitutes the molecular machinery that drives apoptosis. Caspases are selectively activated in a stimulus specific and tissue-specific fashion. The aims of this study were to (1) identify specific caspases activated by glutamine starvation and (2) determine whether a general caspase inhibitor blocks glutamine starvation-induced apoptosis in intestinal epithelial cells. Rat intestinal epithelial (RIE-1) cells were deprived of glutamine. Specific caspase activation was measured using fluorogenic substrate assay. Apoptosis was quantified by DNA fragmentation and Hoechst nuclear staining. Glutamine starvation of RIE-1 cells resulted in the time dependent activation of caspases 3 (10 hours) and 2 (18 hours), and the induction of DNA fragmentation (12 hours). Caspases 1 and 8 remained inactive ZVAD fluoromethyl ketone, a general caspase inhibitor, completely blocked glutamine starvation-induced caspase activation, DNA fragmentation, and nuclear condensation. These results indicate that glutamine starvation selectively activates specific caspases, which leads to the induction of apoptosis in RIE-1 cells. Furthermore, inhibition of caspase activity blocked the induction of apoptosis, suggesting that caspases are potential molecular targets to attenuate apoptotic responses in the gut. PMID- 11058862 TI - Teflon buttress inhibits recanalization of uncut stapled bowel. AB - The uncut Roux limb operation is designed to have the benefits of a Roux limb but still have electrical continuity from proximal to distal bowel, thus eliminating the risk of Roux stasis syndrome. The main complication has been recanalization of the uncut staple line leading to bile reflux. This study aims to employ a new technique, which will not allow recanalization of an uncut staple line but will not interfere with normal bowel myoelectric activity. Fourteen mongrel dogs, 25 to 35 kg, underwent a midline laparotomy under general anesthesia. An uncut staple line was placed 25 cm from the ligament of Treitz. In seven animals an uncut staple line alone was placed, and in the other seven animals the bowel was stapled between a sandwich of Teflon reinforcing strips such that the staples were held on both sides of the bowel by the Teflon. A jejunojejunostomy was placed 6 cm proximal to the staple line. Insulated bipolar electrical leads were placed around the staple line. After the electrical leads were monitored 2 days to 3 months postoperatively for bowel myoelectric activity, The animals were killed and the operative sites inspected. No animal suffered morbidity or mortality from the procedure. All seven unreinforced staple lines recanalized and all seven reinforced staple lines remained competent. The duodenal pacemaker potentials were transmitted through the staple line in five animals (3 controls and 2 with Teflon reinforcement) with in 1 week postoperatively. The uncut staple line does not reliably transmit the duodenal pacemaker potentials. The staple line does not recanalize when it is reinforced with a permanent material, increasing the utility of the "uncut" Roux limb operation. PMID- 11058863 TI - Effect of the distal remnant on ileal adaptation. AB - The ileum has a greater adaptive capacity than the jejunum after intestinal resection, which may be, in part, related to increased exposure to luminal contents and intrinsic properties of the ileum However, the intestinal remnant might contribute to this adaptive response as well. Our aim was to determine the effect of the distal intestinal remnant on ileal adaptation when the ileum is proximal in the intestinal tract. Twenty-one Lewis rats were included in the study. One group (n = 7) served as unoperated control subjects, the second group (n = 7) underwent transposition of the jejunum and ileum, and the third group (n = 7) underwent 50% proximal resection with syngeneic transplantation of the ileum Nutritional status and structural adaptation were studied at 14 days. Animals in both the transposition and transplant groups initially lost weight but weights returned to above preoperative levels at 14 days. Food intake, stool weight, and serum albumin levels were similar in these two groups. Intestinal weight and diameter were similar in the proximal end of the ileal segment in the two study groups and were significantly increased compared to control values (0.26 +/- 0.04 and 0.31 +/- 0.02 vs. 0.10 +/- 0.0 g/cm and 8.4 +/- 0.5 and 9.1 +/- 0.8 vs. 4.9 +/- 0.3 mm; P < 0.05) Intestinal weight and diameter of the distal end of the ileal segment were greater than those values in unoperated control animals but were greatest in the ileal transplant group (0.15 +/- 0.1 and 0.24 +/- 0.03 vs. 0.07 +/- 0.01 g/cm and 5.6 +/- 1.1 and 8.7 +/- 0.6 vs. 4.3 +/- 0.2 mm; P < 0.05). Villus height and crypt depth were similar in both the proximal and distal ends of the ileal segments in the two study groups and were significantly increased compared to control values (642 +/- 75 and 720 +/- 15 vs. 411 +/- 24 proximal and 443 +/- 49 and 500 +/- 46 vs. 343 +/- 22 microm distal, P < 0.05; 223 +/- 34 and 244 +/- 33 vs. 173 +/- 20 proximal and 192 +/- 28 and 209 +/- 18 vs. 144 +/- 26 microm distal, P < 0.05). Proximal placement of the ileum by either transposition or transplantation results in structural adaptation. This occurs to a similar extent whether the distal remnant is jejunum or ileum. Thus increased exposure to luminal contents and intrinsic properties appear to be the important factors in the adaptive capability of the ileum when the ileum is the proximal portion of the intestinal tract. PMID- 11058864 TI - Effects of glutamine isomers on human (Caco-2) intestinal epithelial proliferation, strain-responsiveness, and differentiation. AB - Enteral feeding with small amounts to stimulate bowel motility, and glutamine supplementation, which provides nutrients selectively used by intestinal epithelial cells, might preserve the gut mucosa during fasting. We evaluated the effects of the interaction between mechanical strain and glutamine supplementation in human Caco-2 intestinal epithelial cells, and pursued the finding of equivalent effects of L- and D-glutamine in Caco-2, HT-29, and primary malignant human colonocytes. Caco-2 cells were subjected to repetitive strain in media containing 2 mmol/L of L-glutamine and media supplemented with L- or D glutamine. Proliferation was determined by automated cell counting. Differentiation and cellular production of L-glutamine were determined spectroscopically. Rhythmic deformation stimulated Caco-2 proliferation in a frequency-dependent manner. Maximal stimulation occurred at 10 cpm, consistent with in vivo frequencies of peristalsis and villous motility. Deformation at 10 cpm and L-glutamine supplementation from 2 to 5 mmol/L concentrations independently stimulated Caco-2 proliferation; the combination further increased proliferation. D-Glutamine supplementation yielded similar results, although with lesser potency. Furthermore, both L- and D-glutamine equivalently reduced Caco-2 dipeptidyl dipeptidase activity. The effects of each isoform were blocked by 1 to 3 mmol/L acivicin, a selective antagonist of glutamine metabolism. Indeed Caco-2 and HT-29 cells and primary malignant colonocytes each metabolized D-glutamine to L-glutamine. Glutamine supplementation in fasting patients might prove synergistic with stimulation of bowel motility by non-nutritive feeding, whereas tissue-specific variations in D-glutamine metabolism might facilitate selective nutripharmaceutical targeting of the gut mucosa. PMID- 11058865 TI - Enhancement of metastatic properties of pancreatic cancer cells by MUC1 gene encoding an anti-adhesion molecule. AB - MUC1 mucin expression has been shown to be associated clinicopathologically with metastasis and poor clinical outcome in a variety of tumors. To further investigate this finding experimentally, human pancreatic cancer S2-013 cells overexpressing MUC1 were used for spontaneous metastatic potential in nude mice. It was found that the number of lung metastases of MUC1 transfectants was significantly higher than that of control cells. To analyze the molecular mechanisms that underlie the increased metastatic activity, in vitro adhesion assays were performed. MUC1 mucin expression enhancedin vitro invasiveness and motility of S2-013 cells, and decreased the binding of S2-013 cells to type I collagen, Type IV collagen and laminin. Similar effects were not observed for cells expressing tandem repeat-deleted MUC1 cDNA. Adhesion properties were abolished by benzyl-alpha-GalNAc treatment, indicating that glycosylation of the extracellular domain of MUC1 was essential for these biological adhesive functions. Our data support the hypothesis that MUC1 expression contributes to the metastatic ability of pancreatic cancer cells. PMID- 11058866 TI - Differential expression and subcellular localization of murine BRCA1 and BRCA1 delta 11 isoforms in murine and human cell lines. AB - BRCA1 mutations are involved in breast and ovarian cancer predisposition in humans. The biological functions of the murine BRCA1 gene have been extensively studied but little is known about murine BRCA1 proteins. To better characterize these proteins, we have cloned the full-length murine BRCA1 cDNA and a splice variant deleted of exon 11, BRCA1-delta 11, by RT-PCR method. Three polyclonal antibodies raised against various parts of murine BRCA1 were used in our study: D16, M20 and 5MO, which were generated in our laboratory. This allowed us to analyze the expression and subcellular localization of both isoforms in murine and human cell lines by immunoblotting, immunoprecipitation, cell fractionation and immunofluorescence. Endogenous BRCA1 was detected in murine cell lines but not splice variant BRCA1-delta 11, whereas both ectopically expressed murine isoforms were detected in transfected human Bosc 23 cells. Subcellular fractionation and immunofluorescence results showed that the BRCA1 protein was mainly located in the nucleus, whereas BRCA1-delta 11 was preferentially cytoplasmic. The conservation of exon 11 splicing and the differential subcellular localization of BRCA1 and BRCA1-delta 11 in human and mouse suggest that these proteins could play distinct roles and that they could differentially act in the pathological mechanisms leading to the development of breast and ovarian cancer. The characterization of the murine BRCA1 proteins and antibodies will be useful to further study BRCA1 functions in murine models. PMID- 11058867 TI - Uteroglobin reverts the transformed phenotype in the endometrial adenocarcinoma cell line HEC-1A by disrupting the metabolic pathways generating platelet activating factor. AB - Uteroglobin, originally named blastokinin, is a protein synthesized and secreted by most epithelia, including the endometrium. Uteroglobin has strong anti inflammatory properties that appear to be due, at least in part, to its inhibitory effect on the activity of the enzyme phospholipase A(2). In addition, recent experimental evidence indicates that uteroglobin exerts antiproliferative and antimetastatic effects in different cancer cells via a membrane receptor. The human endometrial adenocarcinoma cell line HEC-1A does not express uteroglobin. Thus, we transfected HEC-1A cells with human uteroglobin cDNA. The transfectants showed a markedly reduced proliferative potential as assessed by impaired plating efficiency as well as by reduced growth in soft agar. Cytofluorimetric analysis clearly indicated that in uteroglobin-transfected cells the time for completion of the cell cycle was increased. We previously demonstrated that HEC-1A cells actively synthesize platelet-activating factor, one of the products of phospholipase A(2) activity. In addition, we demonstrated that platelet activating factor stimulates the proliferation of these cells through an autocrine loop. In uteroglobin transfectants, the activity of phospholipase A(2) and platelet-activating factor acetyl-transferase, which are involved in the synthesis of platelet-activating factor, was significantly reduced compared with wild-type and vector-transfected cells (p < 0.05). Our results indicate that enforced expression of uteroglobin in HEC-1A cells markedly reduced their growth potential and significantly impaired the synthesis of platelet-activating factor, an autocrine growth factor for these cells. These data suggest that one possible mechanism for the recently observed antineoplastic properties of uteroglobin may be the inhibition of the synthesis of platelet-activating factor. PMID- 11058868 TI - Identification of differentially expressed genes in human melanoma cells with acquired resistance to various antineoplastic drugs. AB - Malignant melanoma displays strong resistance against various antineoplastic drugs. The mechanisms conferring this intrinsic resistance are unclear. To better understand the molecular events associated with drug resistance in melanoma, a panel of human melanoma cell variants exhibiting low and high levels of resistance to 4 commonly used drugs in melanoma treatment, i.e., vindesine, etoposide, fotemustine and cisplatin, was characterized by differential display reverse transcription-polymerase chain reaction (DDRT-PCR). Of 269 mRNA fragments found to be altered in expression level by DDRT-PCR, a total of 11 cDNA clones was characterized after confirmation of a differential expression pattern by Northern blot analyses. These clones include 3 genes (DSM-1, DSM-3 and DSM-5) of known function, 4 previously sequenced genes (DSM-2, DSM-4, DSM-6 and DSM-7) of uncharacterized function and 4 novel genes (DSM-8-DSM-11) without match in GenBank. All of these genes exhibited altered mRNA expression in high level etoposide-resistant cells, whereby 7 genes (DSM-1-DSM-6 and DSM-8) were found to be decreased in the transcription rate in these etoposide-resistant cells. The mRNA synthesis of the remaining genes (DSM-7 and DSM-9-DSM11) was enhanced in high level etoposide-resistant melanoma cells. The expression of 5 (DSM-5 and DSM 7-DSM-10) of the cloned cDNA encoding mRNAs was modulated in various independently established drug-resistant melanoma cells, indicating to be associated with drug resistance. Further characterization of these genes may yield inside into the biology and development of drug resistance in malignant melanoma. PMID- 11058869 TI - Expression of a catalytically inactive H118Y mutant of nm23-H2 suppresses the metastatic potential of line IV Cl 1 human melanoma cells. AB - Nm23-H1 and nm23-H2 are putative metastasis suppressor genes that encode nucleoside diphosphate kinase (NDPK) A and B. NDPKs form oligomers distributed between soluble and particulate fractions of cells and therefore may exert their effects as either soluble or bound proteins. To determine whether metastasis related functions of NDPKs are mediated by their catalytic activity in membrane bound or soluble complexes, we have stably transfected highly metastatic human melanoma Line IV Cl 1 cells with wild-type and catalytically inactive (H118Y) nm23-H1 and nm23-H2 genes and assayed their metastatic potential in nude mice. Transfection with wild-type nm23-H1 and nm23-H2 genes and catalytically inactive nm23-H1 did not significantly (all p > 0.10) alter the metastatic potential of Line IV Cl 1 cells while transfection with catalytically inactive nm23-H2 significantly (p < 0.01) reduced their metastatic potential. The lack of effect of transfection with wild-type and catalytically inactive nm23-H1 suggests that neither soluble nor membrane bound NDPK A affect the metastatic potential of Line IV Cl 1 cells. The metastasis suppressive effect of catalytically inactive NDPK B overexpression suggests that competition with bound complexes containing catalytically active NDPK B inhibits metastasis of Line IV Cl 1 cells. These results imply that bound NDPK B promotes metastasis and suggest that inhibition of its function or of its binding to critical sites may be a useful approach to limit the development of metastases in human melanoma. PMID- 11058870 TI - CDKN2A/p16 inactivation in the prognosis of oligodendrogliomas. AB - The cell-cycle regulator p16 inhibits the complex cdk4-cyclin D1 and controls G1 S transition. In human tumors, p16 inactivation is often accomplished by homozygous deletion (HD) of its encoding gene, CDKN2A. Methylation of the 5' CpG island promoter has been proposed as an alternative mechanism of inactivation. Expression of p16, CDKN2A HD and 5' CDKN2A CpG island methylation was studied in 25 oligodendrogliomas by immunohistochemistry and PCR amplification. Ten oligodendrogliomas were p16-immunonegative, and CDKN2A HD was determined in 8 of these cases. In the 2 immunonegative cases without HD, no CpG island methylation was found. The absence of CpG island methylation in the p16-immunonegative cases without HD suggests either non-genetic regulation of p16 or different genetic changes. CDKN2A HD did not correlate with histological grading (p = n.s.); however, it showed a correlation with survival (p = 0.03), supporting an important role of CDKN2A in the prognosis of oligodendrogliomas. PMID- 11058871 TI - Peritoneal colonization by human pancreatic cancer cells is inhibited by antisense FUT3 sequence. AB - Several alpha(1,3/1,4) fucosyltransferases expressed in human pancreatic cancer cells can participate in the biosynthesis of cell surface sialyl-Lewis a and sialyl-Lewis x antigens that contribute to hematogenous metastatis. Previously, we observed a significant increase of the alpha(1,4) fucosyltransferase activity in tumoral pancreatic cell lines, suggesting that FUT3 could be involved in the sialyl-Lewis antigen expression. Therefore, we invalidated the expression of FUT3 by expressing FUT3 antisense sequence in the human pancreatic tumor BxPC-3 cell line, which expresses the alpha(1,4) fucosyltransferase activity and harbors the cell surface sialyl-Lewis antigens. The decrease of FUT3 transcript after transfection of antisense cDNA of FUT3 in these cells results in a substantial reduction of sialyl-Lewis antigen expression on cell surface. This decreased antigen expression was associated with an inhibition of adhesive properties to E selectin and a decrease of metastatic power of FUT3 antisense-transfected BxPC-3 cells as tested in nude mice. Our study provides evidence that the expression level of FUT3 may regulate the expression of sialyl-Lewis a and sialyl-Lewis x surface antigens and consequently could play an important role in metastatic properties of human pancreatic cancer cells. PMID- 11058872 TI - Anti-sense suppression of epidermal growth factor receptor expression alters cellular proliferation, cell-adhesion and tumorigenicity in ovarian cancer cells. AB - Over-expression of epidermal growth factor receptor (EGFR) in ovarian cancer has been well documented. Human NIH:OVCAR-8 ovarian carcinoma cells were transfected with an expression vector containing the anti-sense orientation of truncated human EGFR cDNA. EGFR anti-sense over-expression resulted in decreased EGFR protein and mRNA expression, cell proliferation and tumor formation in nude mice. In accordance with the reduced levels of EGFR in EGFR anti-sense-expressing cells, tyrosine phosphorylation of EGFR was decreased compared to untransfected parental cells treated with EGF. In EGFR anti-sense-transfected cells, expression of erbB-3, but not erbB-2, was increased. In addition, basal and heregulin-beta 1 stimulated tyrosine phosphorylation of erbB-3 was higher in EGFR anti-sense vector-transfected cells. A morphological alteration in EGFR anti-sense gene expressing cells was correlated with a decrease in the expression of E-cadherin, alpha-catenin and, to a lesser extent, beta-catenin. Changes in the expression of these proteins were associated with a reduction in complex formation among E cadherin, beta-catenin and alpha-catenin and between beta-catenin and EGFR in EGFR anti-sense-expressing cells compared to sense-transfected control cells. These results demonstrate that EGFR expression in ovarian carcinoma cells regulates expression of cell adhesion proteins that may enhance cell growth and invasiveness. PMID- 11058873 TI - Prognostic impact of tissue inhibitor of matrix metalloproteinase-1 in esophageal carcinoma. AB - Tissue inhibitor of metalloproteinase-1 (TIMP-1) inhibits the activity of matrix metalloproteinase, which may play an important role in carcinoma invasion and metastasis. TIMP-1 is thus considered to inhibit carcinoma invasion and metastasis. However, TIMP-1 possesses another important function, cell growth promotion. The clinical significance of TIMP-1 expression has not been fully determined in esophageal carcinoma. We thus examined the expression of TIMP-1 mRNA in tumor (T) and corresponding normal (N) tissues of 85 esophageal carcinoma cases by RT-PCR. The T:N ratio of TIMP-1 mRNA expression in each case was evaluated semi-quantitatively with adjustment by an internal control gene. The mean T:N ratio was 2.0 (range 0.2-6.5). When comparing high-expression cases (T:N > 2.0, n = 37) with low-expression cases (T:N < or = 2.0, n = 48), the former showed a significantly higher frequency of lymph vessel invasion, vascular vessel invasion, lymph node metastasis and advanced-stage disease. The former cases showed a poorer prognosis than the latter. Multivariate analysis disclosed that TIMP-1 expression status was an independent determining factor for prognosis. Our findings suggest that TIMP-1 expression correlates with tumor extension of esophageal carcinoma and might, if validated, prove useful as a novel prognostic marker for esophageal carcinoma. PMID- 11058874 TI - Characterization of a mutant E-cadherin protein encoded by a mutant gene frequently seen in diffuse-type human gastric carcinoma. AB - The cell-cell adhesion molecule E-cadherin plays an essential role in the maintenance and function of epithelial tissues. Altered expression of E-cadherin has been implicated in tumor invasion. Analysis of mutations of the human E cadherin gene in gastric carcinoma of the diffuse type has revealed that deletion of exon 8 or 9 in its cDNA appears to be predominant. In this study, we carried out structural and functional analyses of a mutant form of E-cadherin in a cell line, HSC45-M2, established from a human signet ring-cell carcinoma. Although immunohistochemical analysis showed that the mutant cadherin was localized at cell-cell contact sites as usually seen with the wild type, these cells did not form compact colonies. HSC45-M2 cells expressed aberrant E-cadherin with an m.w. larger than that of the wild type. In these cells, we found deletion of the exon 9-intron 9 boundary including the splicing donor site in E-cadherin genomic DNA. RT-PCR indicated 2 transcripts, which appeared to be caused by the splicing defect. Northern blotting, however, showed that the transcript lacking exon 9 was predominantly detected in these cells. The electrophoretic mobilities on SDS-PAGE of the mutant E-cadherin protein in HSC45-M2 cells and the protein expressed from cDNA lacking exon 9 appeared identical. Analysis of the amino-terminal region of the mutant E-cadherin protein revealed that the cadherin was capable of becoming a mature form by removal of its amino-terminal peptide. However, the mutant E cadherin was susceptible to trypsinization in the presence of Ca(2+), which is not the case for wild-type E-cadherin, suggesting that the mutant E-cadherin frequently found in diffuse-type gastric carcinoma may have lost its Ca(2+) binding ability, leading to disruption of the tight cell-cell association. PMID- 11058875 TI - Expression and regulation of neuropilin-1 in human astrocytomas. AB - Vascular endothelial growth factor (VEGF), through activation of its endothelial receptors VEGFR-1 and VEGFR-2, is an important positive modulator of tumor angiogenesis and edema in solid tumors such as malignant astrocytomas. Neuropilin 1 (Npn-1) is a transmembrane receptor expressed by both endothelial and non endothelial cells, including tumor cells. Npn-1 has been postulated to function as a co-factor in activation of the biologically relevant VEGFR-2, by the most abundant VEGF165 isoform. However, the function of Npn-1 in normal and pathological angiogenesis, its expression pattern in relation to VEGF in tumors such as astrocytomas and whether it is similarly or differentially regulated compared to VEGF remain unknown. In our study, the expression pattern of Npn-1 and VEGF by human astrocytoma cell lines and specimens was closely correlated and associated with malignant astrocytomas. Mitogens, such as epidermal growth factor and activation of p21-Ras, previously demonstrated to be relevant in astrocytoma proliferation and induction of VEGF, also induce Npn-1 expression. Hypoxia, the main physiological inducer of VEGF expression, decreased Npn-1 expression. Increased Npn-1 expression was also demonstrated in a transgenic mouse astrocytoma model. Astrocytomas are an ideal system for furthering our understanding of the functional relevance, if any, of Npn-1 in tumor angiogenesis. PMID- 11058876 TI - Extracellular lipid-mediated signaling in tumor-cell activation and pseudopod protrusion. AB - We have pioneered an in vitro pseudopod-generation model wherein suspended tumor cells are stimulated to form pseudopods into glass micropipettes in response to soluble collagen type IV (CIV). Pertussis toxin and removing intracellular calcium were found previously to be inhibitory to that process. We now extend those observations to dissect the roles of transmembrane calcium influx and circulating fatty acids on pseudopod extension. Removal of fatty acids from BSA in basal media resulted in abrogation of pseudopod formation, while reconstitution of free fatty acids restored cell pseudopod protrusion. We thus hypothesized that fatty acids may provide necessary pseudopod stimulatory signals. Addition of lysophosphatidic acid (LPA) to the fatty acid-free CIV solution or in an opposite pipette without CIV permitted approximately 50% pseudopod recovery in all pipette directions in a dose-dependent fashion. Thapsigargin (TG), an agent that releases internal calcium stores and causes opening of store-operated calcium channels, restored pseudopod protrusion up to 80% in CIV with fatty acid-free albumin. [Ca(2+)](i) release was non-additive when cells were stimulated by TG and LPA, suggesting overlapping [Ca(2+)](i) stores. The combination of TG and LPA in fatty acid-free albumin fully restored the pseudopod response to CIV. Addition of EGTA to chelate stimulatory media calcium blocked the pseudopod response to CIV in the presence of fatty acids. This indicates that pseudopod protrusion requires transmembrane calcium entry. Thus, extracellular lipids and calcium mobilization are required to complement CIV in pseudopod protrusion from suspended cells. PMID- 11058877 TI - Polymorphisms of the DNA repair gene XRCC1 and risk of gastric cancer in a Chinese population. AB - Gastric cancer remains the leading cause of cancer death in China and other countries in eastern Asia. Studies of gastric cancer have revealed that it is a disease of complex etiology involving dietary, infectious, environmental, occupational and genetic factors. DNA repair capacity has been suggested as a genetic factor contributing to variation in susceptibility to cancer. In the present study, we described an association between 2 polymorphisms of the DNA repair gene XRCC1 and risk of gastric cancer in a Chinese population. We used a polymerase chain reaction-based assay to detect Pvu II and Nci I restriction fragment length polymorphisms (XRCC1 26304 C-->T and XRCC1 28152 G-->A, respectively) in 188 patients with gastric cancer and 166 healthy controls. The XRCC1 26304 T allele (194Trp) frequency (34.6%) was higher and the XRCC1 28152 A allele (399Gln) frequency (25.6%) was lower in healthy Chinese controls than previously reported healthy U.S. Caucasian controls (7.2% and 34.1%, respectively). Multivariate logistic regression analysis revealed that the putative high-risk genotypes XRCC1 26304 CC and XRCC1 28152 GA/AA were associated with a non-significant increased risk for gastric cancer (adjusted odds ratio [OR]=1.45, 95% confidence interval [CI]= 0.93-2.25 and OR=1.53, 95% CI= 0.98 2.39, respectively) compared with other genotypes. However, the XRCC1 26304 CC genotype was associated with a significantly increased risk for gastric cardia cancer (adjusted OR=1.86, 95% CI=1.09-3.20). Individuals with both putative high risk genotypes (CC and GA/AA) had a significantly higher risk (adjusted OR=1.73, 95% CI=1.12-2.69), particularly for gastric cardia cancer (adjusted OR=2.18, 95% CI=1.21-3.94) than individuals with other genotypes. These findings support the hypothesis that these 2 XRCC1 variants may contribute to the risk of developing gastric cancer, particularly gastric cardia cancer. PMID- 11058878 TI - Comparative genomic hybridization of fine needle aspirates from breast carcinomas. AB - Detailed knowledge of chromosomal aberrations in a specific tumor may facilitate the development of individually tailored chemotherapy, hormone or gene therapy. Unfortunately, karyotype analysis requires living cells and is complicated by the low number of good metaphase spreads obtained. Comparative genomic hybridization (CGH), however, is capable of detecting and mapping genome-wide amplifications and deletions using an equimolar mixture of normal and tumor cell DNA. We show here that even the few cells from a fine needle aspirate of a tumor are sufficient for a direct CGH assay, independent of DNA amplification. Ten primary breast cancers were analyzed by CGH. A fresh frozen fine needle aspirate and a formalin-fixed and paraffin-embedded section were used for each tumor. Metaphases from each CGH reaction were imaged, and a sum ratio profile was determined for every chromosome. The ratio profiles of DNA isolated from the 2 material sources were then compared. Fine needle aspirates and the paraffin-embedded material of a single tumor yielded the same fluorescence ratio profiles, albeit with slightly different confidence intervals. Different tumors showed a variety of aberrations. The most frequently observed changes were 1q+, 8q+, 14q-, 16p+, 16q-, 17p-, 17q+, 19q+, 20q+, 21q- and 22q-. The ability of CGH to assess chromosomal changes in breast cancer from fine needle aspirates could facilitate genetic evaluation of tumors prior to surgery. PMID- 11058879 TI - Genetic alterations in DNA diploid, aneuploid and multiploid colorectal carcinomas identified by the crypt isolation technique. AB - Loss of heterozygosity (LOH) and microsatellite instability (MSI) commonly occur in colorectal carcinomas. However, the role of these genetic alterations in determining DNA ploidy status of tumors (diploid, aneuploid and multiploid) remains unclear. In the present study, we attempted to clarify the relationship between genetic alterations and DNA ploidy status. Crypt isolation coupled with DNA cytometric sorting and polymerase chain reaction assay (17 microsatellite markers) were used to study allelic losses and MSI in 59 colorectal carcinomas (diploid, 15; aneuploid, 10 and multiploid, 34). Of the 15 diploid carcinomas, 6 exhibited MSI in which allelic losses were rarely found. The other 9 diploid tumors mostly exhibited allelic losses, but none displayed MSI status. Whereas allelic losses frequently occurred in the aneuploid carcinomas and the aneuploid populations of multiploid carcinomas, they were rarely detected in the diploid populations of multiploid carcinomas. MSI status was not observed in aneuploid carcinomas nor in either population of multiploid carcinomas. Although multiploid carcinomas genetically resemble aneuploid carcinomas in the expression of the severe LOH phenotype, the genetic alterations seen in the diploid populations of multiploid carcinomas may differ from those of diploid carcinomas. Furthermore, all diploid, aneuploid and both the diploid and aneuploid fractions of the multiploid tumors that were non-MSI exhibited a high rate of LOH, suggesting that LOH is independent of the tumor's ploidy status. PMID- 11058880 TI - Molecular analysis of PTEN and MXI1 in primary bladder carcinoma. AB - Loss of heterozygosity (LOH) on 10q is associated with late-stage events in urothelial neoplastic progression. The tumor suppressor gene PTEN, which is mutated or homozygously deleted in numerous cancers, maps to a region of 10q within the reported region of minimal loss in bladder tumors. In two recent studies alterations in the PTEN gene occur at a low frequency in bladder tumors displaying 10q LOH. We have screened 35 late-stage bladder tumors for mutations in PTEN and MXI1, both genes mapping to chromosome 10q. Using single-strand conformation polymorphism analysis, we identified 6 tumors harboring mutations in PTEN and 2 additional tumors displaying homozygous deletion at this locus. No MXI1 mutations were identified within the same tumor panel. Of 16 bladder tumor cell lines analyzed, 2 showed homozygous deletion of PTEN and 3 harbored point mutations resulting in an amino acid change. Two cell lines harbored missense mutations in MXI1. We report a significantly higher frequency of PTEN alterations in bladder carcinoma (23%) than was previously recorded, with no accompanying mutations in the MXI1 gene. PMID- 11058881 TI - Sequences of cytotoxic T-lymphocyte epitopes in the Epstein-Barr virus (EBV) nuclear antigen-3B gene in a Japanese population with or without EBV-positive lymphoid malignancies. AB - Latent infection antigens of EBV, including EBV nuclear antigens (EBNAs) and latent membrane proteins, are expressed in latently infected and immortalized B cells but work as target antigens for host cytotoxic T-lymphocyte (CTL) responses in an HLA class I-restricted manner. Among these latent antigens, the immunodominant CTL epitopes in EBNA3B (EBNA3B 399-408 and EBNA3B 416-424) are well characterized. Mutations and strain differences in these sequences, compared to the prototype A sequence, reduce CTL responses to latently infected B cells. These EBNA3B CTL epitopes in the normal Japanese population and in 2 lymphoid neoplasias, pyothorax-associated lymphoma (PAL) and nasal natural killer-cell lymphoma, were directly sequenced by PCR. Most EBV in peripheral blood leukocytes (PBLs) from healthy Japanese donors exhibited the prototype A sequence, with mutations in approximately 20% (3/16). The sequence of EBNA3B CTL epitopes in lymphoma tissue was obtained in 6 PAL cases, and 5 exhibited mutations or strain differences compared to the prototype A sequence. Furthermore, the EBNA3B sequence in PAL tissue was different from that in PBLs of the same patient or 1 of the sequences found in PBLs. However, the EBNA3B gene in nasal lymphoma tissues exhibited predominantly the prototype A sequence. Because PAL cells expressed EBNA3B mRNA, detected by RT-PCR, but nasal lymphoma cells did not, mutations and strain differences of the sequences of EBNA3B CTL epitopes were specific findings in EBNA3B-positive lymphomas. PMID- 11058882 TI - Identification of SART3-derived peptides capable of inducing HLA-A2-restricted and tumor-specific CTLs in cancer patients with different HLA-A2 subtypes. AB - We recently identified the SART3 antigen encoding shared tumor epitopes recognized by HLA-A2402-restricted and tumor-specific CTLs. Our study investigated whether the SART3 antigen encodes peptides recognized by the HLA-A2 restricted CTLs. The HLA-A2-restricted and tumor-specific CTL line recognized COS 7 cells co-transfected with the SART3 gene and either HLA-A0201, -A0206 or -A0207 cDNA but not those co-transfected with the SART3 gene and HLA-A2402 or -A2601 cDNA. The 2 SART3 peptides at positions 302 to 310 and 309 to 317 possessed the ability to induce HLA-A2-restricted and tumor-specific CTLs from peripheral blood mononuclear cells of cancer patients with various histological types and different HLA-A2 subtypes. Therefore, these 2 peptides could be useful for specific immunotherapy of a relatively large number of HLA-A2(+) cancer patients. PMID- 11058883 TI - Enhancement of antitumor effect of bleomycin by low-voltage in vivo electroporation: a study of human uterine leiomyosarcomas in nude mice. AB - Uterine leiomyosarcoma is an extremely malignant neoplasm with high rates of distant metastasis, and systemic chemotherapy is not particularly effective. Thus, the introduction of more active anticancer agents, or of a new drug delivery system, is urgently needed. Recently, electrochemotherapy has been introduced as a way of enhancing the cytotoxic effects of chemotherapeutic agents. This involves administering the drug in combination with electric pulses (which permeabilize tumor cell membranes and allow the drug to enter the cells). In particular, bleomycin (BLM) cannot cross the plasma membrane efficiently, but its cytotoxicity can be enhanced by electropermeabilization. The aim of the present study was to investigate the effect of low-voltage electroporation (EP) in combination with local BLM injection on the growth of uterine leiomyosarcoma in nude mice. Human uterine leiomyosarcoma cells (SK-LMS-1) were implanted subcutaneously into nude mice. Tumor growth in mice treated with EP (100 V/cm) plus BLM was compared with that in mice receiving BLM alone, EP alone, or no treatment (controls). Tissue BLM concentrations and histological analysis (including mitotic counts) were evaluated in tumor tissues. There was a significant reduction in tumor growth in mice that received EP with BLM. One hour after the treatment, the local BLM concentration was 10 times higher in the tumors that received EP with BLM than in those receiving only BLM. Moreover, the mitotic count was lower in the tumors that received EP plus BLM than in the controls. These results demonstrate the possible therapeutic value of low-voltage EP with BLM in human uterine leiomyosarcoma. PMID- 11058884 TI - Intra-tumoral injection of doxorubicin (adriamycin) encapsulated in liposome inhibits tumor growth, prolongs survival time and is not associated with local or systemic side effects. AB - Encapsulation of doxorubicin (Adriamycin) in liposome (LipADM) augments the anti tumor effects of the drug and reduces side effects such as cardiotoxicity. However, it does not always enhance anti-tumor effects because of entrapment by the reticuloendothelial system. In this study, we investigated the anti-tumor effect of LipADM injected directly into the tumor to augment tumor targeting. LipADM (7.5 mg/kg body weight), the same concentration as free ADM (FADM), was injected percutaneously or i.v. into 7-day-old established Meth-A tumors in mice. Mock liposome was injected percutaneously into tumors of control mice. Mean relative tumor weights of the 5 groups on day 15 were as follows: intra-tumoral injection of LipADM, 2.92 +/- 1.09; intra-tumoral injection of FADM, 6.99 +/- 2.92; i.v. injection of LipADM, 11.07 +/- 7.95; i.v. injection of FADM, 11.80 +/- 6.55; control, 23.94 +/- 9.03. Mean survival times were as follows: intra-tumoral injection of LipADM, 46.2 +/- 11.0 days; FADM, 34.6 +/- 9.6 days; mock control, 30.2 +/- 4.8 days. Histological examination showed no tissue damage at the site of s.c. injection of LipADM. ADM concentrations in tumor tissues after intra tumoral injection were persistently high in the LipADM-treated group. Our results indicate that direct injection of LipADM into the tumor is therapeutically useful by producing persistently high concentrations of ADM in the target tissue, with few local and systemic side effects. PMID- 11058885 TI - In vivo inhibition of PC-3 human androgen-independent prostate cancer by a targeted cytotoxic bombesin analogue, AN-215. AB - The effectiveness of chemotherapy targeted to bombesin (BN) receptors was evaluated in nude mice bearing PC-3 human androgen-independent prostate cancers. Cytotoxic BN analogue AN-215, consisting of 2-pyrrolinodoxorubicin (AN-201) linked to BN-like carrier peptide RC-3094, was injected i.v. at 150 nmol/kg on days 1, 11 and 21. After treatment with AN-215, tumor volume was 69% (p < 0.01) smaller than that in controls and tumor doubling time was extended from 8.5 +/- 0.7 days to 20.3 +/- 3.5 days (p < 0.05). Cytotoxic radical AN-201, carrier RC 3094 and their unconjugated mixture administered at the same dosage were ineffective. The mortality rate was 12.5% in the AN-201 group and 16.7% in the group treated with the mixture, but no deaths occurred in mice receiving AN-215. Because the ester bond linking AN-201 to the carrier molecule is hydrolyzed much faster in mouse serum than in human serum, in the second experiment we investigated the tolerance to AN-215 and its effect in nude mice bearing PC-3 tumors after pharmacological inhibition of serum carboxylesterases. Two applications of AN-201 at 200 nmol/kg were lethal, whereas no mortality was observed after 4 injections of AN-215 at the same dose. Administration of 200 nmol/kg AN-215 on days 1, 7, 17 and 26 again produced 69% tumor inhibition. BN receptors on membranes of PC-3 tumors were detected by (125)I-[Tyr(4)]BN binding, and expression of mRNA for BRS-3 and GRP-R subtypes was also found. AN-215 showed a high affinity to PC-3 tumors, displacing the radioligand at an IC(50) of 12.95 +/- 0.35 nM. Because BN receptors are present on primary and metastatic prostate cancer, targeted chemotherapy with AN-215 might benefit patients with advanced prostatic carcinoma who relapsed androgen ablation. PMID- 11058886 TI - Influence of mate drinking, hot beverages and diet on esophageal cancer risk in South America. AB - To estimate the effects of consuming hot beverages, including mate (an infusion of the herb Ilex paraguayensis), tea, coffee and coffee with milk, and other food items on esophageal cancer risk, we analyzed data from 830 cases and 1,779 controls participating in a series of 5 hospital-based case-control studies of squamous-cell carcinoma of the esophagus conducted in high-risk areas of South America. After adjusting for the strong effects of tobacco and alcohol consumption, both heavy mate drinking (>1 l/day) and self-reported very hot mate drinking were significantly associated with esophageal cancer risk in men and women. The magnitude and strength of the association for mate amount and, to a lesser extent, mate temperature were higher for women than men. The joint effects of mate amount and mate temperature were more than multiplicative, following a statistically significant synergistic interaction (p = 0.02) which was particularly evident among heavy drinkers (>1.50 l/day) of very hot mate (odds ratio = 4.14, 95% confidence interval: 2.24-7.67) compared to light drinkers (<0.50 l/day) of cold/warm/hot mate. Consumption of other very hot beverages, such as tea and coffee with milk but not coffee alone, was also significantly associated with an increased risk, in the 2- to 4-fold range. Statistically significant protective associations were identified for high consumption of vegetables, fruits, cereals and tea. In contrast, frequent consumption of meat, animal fats and salt was associated with a moderately increased risk. This pooled analysis adds evidence for a carcinogenic effect of chronic thermal injury in the esophagus induced by the consumption of very hot drinks, including mate. Our study further confirms the protective effect of a dietary pattern characterized by daily consumption of fruits and vegetables and low consumption of meat and animal fats. PMID- 11058887 TI - Role of smoking and diet in the cross-cultural variation in lung-cancer mortality: the Seven Countries Study. Seven Countries Study Research Group. AB - We examined the role of smoking and diet in the cross-cultural variation in lung cancer mortality, using aggregated data of the Seven Countries Study, a follow-up study comprising 12,763 middle-aged men in 16 cohorts in Europe, the United States and Japan, which started around 1960. Smoking habits were assessed with a standardised questionnaire. Dietary intake was collected in random sub-samples of each cohort by the dietary record method. Cohort-specific 25-year lung-cancer mortality among all men and among categories of smoking behaviour was related to smoking prevalence and population average dietary intake, respectively, using Poisson regression. Smoking prevalence was positively associated with lung-cancer mortality [risk ratio 1.47, 95% confidence interval (CI) 1.05-2.07, for an increase of 10 percentage points]. Lung-cancer mortality among smokers, which varied significantly among cultures, was positively associated with average fat intake, especially saturated fat intake (rate ratio 1.10, 95% CI 1.04-1.17, for an increase of 4.6 g) but not with unsaturated fat intake. Average fruit and vegetable intake were not related to lung-cancer mortality. Among never-smokers, the power to detect associations was low. In conclusion, both smoking prevalence and average fat intake, especially saturated fat, may play a role in the cross cultural variation in lung-cancer mortality, either independently or by effect modification. PMID- 11058888 TI - Second malignant neoplasms after cancer in childhood and adolescence: a population-based case-control study in the 5 Nordic countries. The Nordic Society for Pediatric Hematology and Oncology. The Association of the Nordic Cancer Registries. AB - Our purpose was to assess the risk of developing a second malignant neoplasm (SMN) after cancer in childhood and adolescence associated with different treatment modalities. Our investigation was performed as a nested case-control study within a Nordic cohort of 25,120 patients younger than 20 years old at first malignant neoplasm (FMN) diagnosed in 1960 through 1987. SMNs were diagnosed in 1960 through 1991. For each case of SMN, 3 controls were sampled, matched by sex, age, calendar year of diagnosis and length of follow-up. For the final analysis, there were 234 cases and 678 controls. Relative risks (RRs) of various exposures were estimated by means of conditional logistic regression, with non-exposed as the reference. The RR of developing SMN in the radiated volume was 4.3 (95% confidence interval 3.0-6.2). The risk was highest in children diagnosed before the age of 5 years; it increased with the dose of radiation and with increasing follow-up time after FMN. Chemotherapy alone was not associated with an increased RR, but it significantly potentiated the effect of radiotherapy. RRs were unchanged between the periods 1960-1973 and 1974-1987, and since the use of chemotherapy increased in the latter period, the number of SMNs may increase. Hereditary factors were important for the occurrence of SMN independently of therapy. We conclude that radiation was the most important treatment-related risk factor for the development of SMN. Chemotherapy appeared to play only an accessory role during the study period, potentiating the carcinogenic effect of radiotherapy. PMID- 11058889 TI - No increased risk of breast cancer after cholecystectomy. AB - An increased risk of breast cancer after cholecystectomy has been proposed, but the results in different studies have not been consistent. To explore this potential association, we conducted a large, population-based, record-linkage, cohort study in Sweden. The cohort consisted of 154,283 women who had been cholecystectomized during the period 1965-1994. The cohort was followed up for breast cancer by the Swedish Cancer Register. The standardized incidence ratio (SIR), the ratio of the observed to the expected number of incident cancers derived from the entire Swedish population, estimated relative risk. During a follow-up of 31 years, we observed 3,879 cases of breast cancer in the cohort vs. 3,843 expected [SIR = 1.01, 95% confidence interval (CI) 0.98-1.04]. No significant difference in risk was observed across duration of follow-up or age at follow-up. Among women followed up for more than 20 years after cholecystectomy, the observed 403 cases of breast cancer rendered an SIR of 0.96 (95% CI 0.87-1.06). The results remained virtually unchanged after controlling for possible confounding effects of obesity and diabetes. Also, we did not find any evidence of an association between gall stone disease per se and the risk of breast cancer. We conclude that cholecystectomy and breast cancer are not likely to be associated. PMID- 11058890 TI - Human bladder cancer, schistosomiasis, N-nitroso compounds and their precursors. PMID- 11058891 TI - Pronucleotides: toward the in vivo delivery of antiviral and anticancer nucleotides. AB - To overcome the many hurdles preventing the use of antiviral and anticancer nucleosides as therapeutics, the development of a prodrug methodology (i.e., pronucleotide) for the in vivo delivery of nucleotides has been proposed as a solution. The ideal pronucleotide should be non-toxic, stable in plasma and blood, capable of being i. v. and/or orally dosed, and intracellularly convertible to the corresponding nucleotide. Although this goal has yet to be achieved, many clever and imaginative pronucleotide approaches have been developed, which are likely to be important pharmacological tools. This review will discuss the major advances and future directions of the emerging field of antiviral and anticancer pronucleotide design and development. PMID- 11058892 TI - Immunophilins: switched on protein binding domains? AB - Peptidylprolyl isomerases (PPIases) are a group of cytosolic enzymes first characterized by their ability to catalyze the cis-trans isomerization of cis peptidylprolyl bonds. Subsequently, some PPIases were also identified as the initial targets of the immunosuppressant drugs-cyclosporin A (CsA), FK506, and rapamycin-have been called immunophilins. Immunophilins have been found to be both widely distributed and abundantly expressed leading to suggestions that they may play a general role in cellular biochemistry. However, the nature of this role has been difficult to elucidate and is still controversial in vivo. A number of roles for these enzymes have been identified in vitro including the ability to catalyze the refolding of partly denatured proteins and stabilize multiprotein complexes such as Ca(2+) channels, inactive steroid receptor complexes, and receptor protein tyrosine kinases. Generally, these effects appear to depend on the ability of immunophilins to selectively bind to other proteins. This review will examine in detail experimental and structural investigations of the mechanism of PPIase activity for both FKBPs and cyclophilins and suggest a mechanism for these enzymes, which depends on their ability to recognize a specific peptide conformation rather than sequence. Examination of structures of immunophilin-protein complexes will then be used to further suggest that the ability of these enzymes to recognize specific peptide conformations is central to the formation of these complexes and may constitute a general function of immunophilin enzymes. The binding of ligand to immunophilins will also be shown to stabilize specific conformations in surface loops of these proteins that are observed to play a critical role in a number of immunophilin-protein complexes suggesting that the immunophilins may constitute a class of ligand-triggered selective protein binders. PMID- 11058893 TI - Applications of Lewis acids for the efficient syntheses of diltiazem, cephems, and taxoids. AB - An efficient method is described for preparation of diltiazem hydrochloride (Herbesser(R)), a marketed calcium antagonist widely used for the treatment of ischemic heart disease. In the reaction of 2-nitrothiophenol (1) with trans-3 phenylglycidic esters (2) carrying various substituents on the benzene ring, both reactivity and stereoselectivity of the oxirane ring-opening of the glycidates were markedly influenced by the electronic nature of the substituents. As a result of our investigation on the catalytic effect of various Lewis acids in the reaction of 2a with 1, tin compounds were found to be effective catalysts for the cis-opening and readily produced the threo-nitro ester (3a-t), a key intermediate for the synthesis of diltiazem. Isolation of the crystalline complex from the reaction of 1 and SnCl(4); and its efficient catalytic activity similar to that of SnCl(4) suggests that the transition state involves co-coordination of tin derivatives both with 1 and the epoxy oxygen of 2a to result in highly specific cis-opening. We have also amplified this chemistry into other fields, leading to applications in the syntheses of cephem and taxoid templates. PMID- 11058894 TI - Hepatocyte nuclear factor 1 alpha (HNF-1 alpha) mutations in maturity-onset diabetes of the young. AB - Maturity-onset diabetes of the young (MODY) is a monogenic form of diabetes mellitus characterized by autosomal dominant inheritance, early age of onset (<25 years) and pancreatic beta-cell dysfunction. MODY is genetically heterogeneous with five different genes identified to date: hepatocyte nuclear factor-4 alpha (HNF-4 alpha) [MODY1]; glucokinase [MODY2]; hepatocyte nuclear factor-1 alpha (HNF-1 alpha) [MODY3]; insulin promoter factor-1 (IPF-1) [MODY4]; and hepatocyte nuclear factor-1 beta (HNF-1 beta) [MODY5]. Mutations in the HNF-1 alpha gene represent a common cause of MODY in the majority of populations studied. Sixty five different mutations have been described in a total of 116 families. The most common mutation is a C-insertion (P291fsinsC) in the polyC tract of exon 4, which has been reported in 22 families. The identification of an HNF-1 alpha gene mutation in a patient with type 2 diabetes confirms the diagnosis of MODY and has important implications for clinical management. PMID- 11058895 TI - Mutations in PMM2 that cause congenital disorders of glycosylation, type Ia (CDG Ia). AB - The PMM2 gene, which is defective in CDG-Ia, was cloned three years ago [Matthijs et al., 1997b]. Several publications list PMM2 mutations [Matthijs et al., 1997b, 1998; Kjaergaard et al., 1998, 1999; Bjursell et al., 1998, 2000; Imtiaz et al., 2000] and a few mutations have appeared in case reports or abstracts [Crosby et al., 1999; Kondo et al., 1999; Krasnewich et al., 1999; Mizugishi et al., 1999; Vuillaumier-Barrot et al., 1999, 2000b]. However, the number of molecularly characterized cases is steadily increasing and many new mutations may never make it to the literature. Therefore, we decided to collate data from six research and diagnostic laboratories that have committed themselves to a systematic search for PMM2 mutations. In total we list 58 different mutations found in 249 patients from 23 countries. We have also collected demographic data and registered the number of deceased patients. The documentation of the genotype-phenotype correlation is certainly valuable, but is out of the scope of this molecular update. The list of mutations will also be available online (URL: http://www.kuleuven. ac.be/med/cdg) and investigators are invited to submit new data to this PMM2 mutation database. PMID- 11058896 TI - PMM2 mutation spectrum, including 10 novel mutations, in a large CDG type 1A family material with a focus on Scandinavian families. AB - Carbohydrate-deficient glycoprotein syndrome type IA (CDG IA) is an autosomal recessive disease characterized clinically by severe involvement of the central and peripheral nervous system, and biochemically by complex defects in carbohydrate residues in a number of serum glycoproteins. CDG IA is caused by mutations in the PMM2 gene located in chromosome region 16p13. In this study, 61 CDG type IA patients (122 chromosomes) were screened for mutations in the PMM2 gene using a combination of SSCP and sequence analysis. More than 95% of the mutations could be detected. All of them were missense mutations. Mutations 422G>A and 357C>A were strikingly more common in the material and comprised 58% of mutations detected. Of the 20 mutations found, 10 were not reported previously. Seven mutations, e.g. 26G>A (five alleles) and 548T>C (seven alleles), were found only in Scandinavian families. The most common genotype was 357C>A/422G>A (36%). Three patients were homozygous, 357C>A/357C>A (two cases), and 548T>C/548T>C (one case). No patients homozygous for the most common mutation 422G>A were detected. The different mutations were clustered e.g., in that most were located in exon 5 (five) and exon 8 (six), while no mutation was detected in exon 2. When the frequencies of each mutation were included, exon 5 comprised 61% (65 chromosomes) of the mutations; in Scandinavian patients the frequency of these mutations was 72%. Thus, analysis of exon five in these patients enables both reliable and time-saving first screening in prenatal diagnostic cases. This could be followed by a second step of additional strategies for the detection of other mutations. PMID- 11058897 TI - Functional analysis of mutations in the OCTN2 transporter causing primary carnitine deficiency: lack of genotype-phenotype correlation. AB - Primary carnitine deficiency is an autosomal recessive disorder of fatty acid oxidation caused by defective carnitine transport. This disease is caused by mutations in the novel organic cation transporter OCTN2 (SLC22A5 gene). The disease can present early in life with hypoketotic hypoglycemia or later in life with skeletal myopathy or cardiomyopathy. To determine whether the variation in phenotypic severity is due to mutations retaining residual function, we extended mutational analysis of OCTN2 to four additional European families with primary carnitine deficiency. Three patients were homozygous for novel missense mutations (R169W, G242V, A301D). The fourth patient was compound heterozygous for R169W and W351R substitutions. Stable expression of all the mutations in CHO cells confirmed that all mutations abolished carnitine transport, with the exception of the A301D mutation in which residual carnitine transport was 2-3% of the value measured in cells expressing the normal OCTN2 cDNA. Analysis of the patients characterized in molecular detail by our laboratory failed to indicate a correlation between residual carnitine transport and severity of the phenotype or age at presentation. PMID- 11058898 TI - Jagged1 (JAG1) mutation detection in an Australian Alagille syndrome population. AB - Alagille syndrome (AGS) is an autosomal dominant disorder characterized by abnormal development of the liver, heart, skeleton, eye, and face. Mutations in the Jagged1 gene (JAG1) have been found to result in the AGS phenotype and both protein truncating mutations and missense mutations have been identified. Using single stranded conformational polymorphism analysis we have screened 22 AGS affected individuals from 19 families for mutations within Jagged1. Twelve distinct Jagged1 mutations were identified in 15 (68.2%) of the 22 AGS cases, seven of which are novel. The mutations include three small deletions (25%), two small insertions (16.6%), three missense mutations (25%), two nonsense mutations (16.6%), and two splice-site mutations (16.6%). These mutations are spread across the entire coding sequence of the gene and most are localized to highly conserved motifs of the protein predicted to be important for Jagged1 function. One-half of the mutations found in this study are located between exons 9 and 12, a region constituting only 12% of the coding sequence. A splice-donor site mutation in intron 11 was shown to cause aberrant splicing of Jagged1 mRNA, consequently terminating translation prematurely in exon 12. The results of this study are consistent with the proposal that either haploinsufficiency for wild type Jagged1 and/or dominant negative effects produced by mutated Jagged1 are responsible for the AGS phenotype. PMID- 11058899 TI - Mutation analysis of the hamartin gene using denaturing high performance liquid chromatography. AB - Denaturing high performance liquid chromatography (DHPLC) is a novel high capacity technique for gene mutation scanning. We have assessed the sensitivity and specificity of this method for analysis of the full coding sequence of the hamartin (TSC1) gene in 20 tuberous sclerosis patients, whose TSC1 genes previously had been studied by single strand conformation polymorphism analysis and protein truncation assay. All eight sequence variants previously identified were adequately detected by DHPLC. Additionally, this approach picked up three polymorphisms, one of which (IVS13-55 C>G) was hitherto unreported, therefore serving as proof of principle for this technique. Thus, DHPLC appears to be a highly sensitive method with advantages in terms of flexibility, fragments size analysis, cost and time and labor sparing, compared to classical approaches of mutation scanning. PMID- 11058900 TI - Hierarchical mutation screening protocol for the BRCA1 gene. AB - The identification of mutations in the BRCA1 gene poses difficulties in achieving a screening outcome that satisfies the twin needs of speed and accuracy. These needs must also take into account the patient's family history and the statistical evaluation of the probability of detecting a mutation. Given the above, we present here a hierarchical mutation screening strategy that comprises two tiers: first, multiplex heteroduplex and exon 13 duplication analysis; second, exon amplification and direct sequencing using a 96-well tray format. The advantages of this strategy are two-fold: first, the division of analytical tools in order to achieve low and high-resolution mutation screening, respectively; second, a streamlined sequencing approach that leads to a sensitive and rapid assay that reduces labor costs and handling errors. The success of this approach is shown by the identification of a novel deletion mutation in exon 14 of the BRCA1 gene, which was not detected by the more conventional protein truncation assay due to the small size of the predicted truncated protein. PMID- 11058901 TI - Diagnosis of haploidy and triploidy based on measurement of gene copy number by real-time PCR. AB - We report the development of a method for diagnosis of heterozygous deletions or duplications based on measurement of gene copy number. The method involves amplifications of a test locus with unknown copy number and a reference locus with known copy number using real-time PCR. Progress of the PCR reactions is monitored using fluorigenic probes and a real-time fluorescence detection system. For each reaction, the number of cycles is measured at which a defined threshold fluorescence emission is reached. Using standard curves, the copy number of the test DNA relative to a common standard DNA is determined for each locus. From the ratio of the relative copy numbers, the genomic copy number of the test locus is determined. In order to demonstrate the accuracy and reliability of the method for genetic testing, we analyzed 43 patients with hereditary neuropathy with liability to pressure palsies (HNPP), containing a heterozygous deletion of a 1.5 Mb region on chromosome 17p11.2-p12, eight patients with Charcot-Marie-Tooth disease, containing a heterozygous duplication of the same genomic region, and 50 normal control individuals. As a test locus we analyzed the PMP22 gene located within the 1.5 Mb region. The genomic copy number of the test locus was precisely measured, and the presence or absence of the genomic deletion or duplication was unambiguously diagnosed in all individuals. PMID- 11058902 TI - Comparative sequence analysis (CSA): a new sequence-based method for the identification and characterization of mutations in DNA. AB - Direct sequencing analysis is largely used to confirm and characterize mutations previously detected by more rapid tests. We have developed a method-Comparative Sequence Analysis (CSA)-that simplifies the analysis of sequencing data facilitating its use as a first screen for mutation detection. Sequence data were split into their component electrophoretograms and the use of a size standard enabled equivalent traces from different individuals to be overlaid. This allowed simple and rapid visual analysis of the results. Using this technique in a blind study, we tested 576 samples for mutations in the Von Hippel-Lindau tumor suppressor gene, VHL. We were able to identify and characterize all 78 known mutations present within the sample set (100% sensitivity and specificity). PMID- 11058903 TI - Genetic heterogeneity of glycogen storage disease type Ia in France: a study of 48 patients. AB - Forty-eight patients with glycogen storage disease type Ia (GSD Ia) were studied. Using a combination of single-strand conformation polymorphism (SSCP) analysis, restriction enzyme digestion and direct sequencing, we were able to identify 93/96 mutant alleles, comprising 23 different mutations in the glucose-6 phosphatase gene (G6PC). Among these, 7 are novel mutations of G6PC: M5R, T111I, A241T, C270R, F322L, and two deletions, 793delG and 872delC, resulting in the same mutation at the amino acid level, fs300Ter (300X). PMID- 11058904 TI - Novel splicing and missense mutations in autosomal dominant polycystic kidney disease 1 (PKD1) gene: expression of mutated genes. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a common disorder mostly characterized by cyst formation in kidney tubules. The majority of ADPKD cases is caused by mutations in the PKD1 gene, but no prevalent mutation has been reported. By heteroduplex analysis of the 3' single-copy region of the gene, we have searched for mutations in subjects from 40 ADPKD families of Northern Italy. Seven novel polymorphisms and three novel disease-associated mutations (R3718Q, L3851P and IVS45+56del25) were identified. Both missense mutations are located in the major extracellular loop of polycystin-1. The 25 bp deletion inside intron 45 did not affect 5' and 3' consensus splicing sites, but caused a 56 nucleotide out of frame-deletion due to activation of a cryptic 3' splice site in exon 46. The mutated RNA should produce a truncated polycystin 1 at the G binding peptide in the intracellular C-terminal end of the protein. RT-PCR analysis showed that the disease-associated mutations were present in transcribed sequences. In particular, RNA analysis of BHK cells transfected with PKD1 genomic DNA, including the deleted intron, showed that no normal transcript is produced by the deleted gene. This intronic mutation, found in a large pedigree, seems to be associated with a prevalence of cerebrovascular disease. PMID- 11058905 TI - Six novel MEN1 gene mutations in sporadic parathyroid tumors. AB - We report nine mutations of the multiple endocrine neoplasia type 1 (MEN1) gene in sporadic parathyroid adenomas. Six of them have not previously been described: E60X, P32R, 261delA, 934+2T-->G, S443P, and 1593insC. The tissue samples were initially submitted to LOH analysis at 11q13 followed by SSCP screening of LOH positive samples. Mutations were identified by direct sequencing and subcloning. Three (E60X, P32R, and 261delA) were in exon 2, one (934+2bp) in the splice junction of exon 5, one (S443P) in exon 9, and one (1593insC) in exon 10. The 3 mutations in exon 2 were associated with loss and/or creation of a restriction site. The corresponding germline sequence of the MEN1 gene was normal. Most mutations would likely result in a nonfunctional menin protein, and therefore in the loss of a tumor suppressor protein. PMID- 11058906 TI - Pharmacogenetics of catechol-O-methyltransferase: frequency of low activity allele in a Ghanaian population. AB - Catechol-O-methyltransferase (COMT) catalyses the O-methylation of neurotransmitters, catechol hormones and drugs such as levodopa and methyldopa. Ethnic differences in COMT activity have been observed in several populations. Previous studies suggest that the g1947G>A low activity allele is less common in individuals of African origin. COMT genotyping was performed using a mini sequencing method in 195 healthy Ghanaians with a frequency of the homozygous g1947G>A of 6%. This study provides confirmation that the low activity COMT allele is less common in individuals of African origin. This finding may be important clinically with regards to the treatment of many neuropsychiatric disorders and in the pathophysiology of various human disorders including estrogen-induced cancers, Parkinson's disease, depression and hypertension. PMID- 11058907 TI - Recurrent and novel mutations of GCDH gene in Chinese glutaric acidemia type I families. AB - Glutaric acidemia type I is caused by mutations of the glutaryl-CoA dehydrogenase (GCDH) gene resulting in loss of GCDH enzyme activity. Patients present with progressive dystonia and lesions in basal ganglia. Dietary treatment, when instituted from the early neonatal period, markedly reduces dystonia and morbidity. Early diagnosis and prenatal diagnosis will be facilitated by knowledge of locally prevalent GCDH mutations. Several common GCDH mutations have been found in different ethnic groups. GCDH mutations were studied in 5 Chinese glutaric acidemia type I families. We detected two novel recurrent mutations (A219T and IVS10-2A>C) which were found in two unrelated families. An asymptomatic carrier of IVS10-2A>C was also found on screening of 120 individuals. Other mutations were identified, including two other novel (R386G & IVS3+1G>A) and two known mutations (G178R & R355H). Fibroblasts from patients carrying the novel mutations were confirmed to be deficient for GCDH activity. This is the first report of GCDH mutations describing recurrent mutations in Chinese patients. The carrier rate of IVS10-2A>C may be particularly high in Chinese. PMID- 11058908 TI - A new case of alpha-1-antitrypsin frameshift mutation (1123insT) causing severe deficiency and emphysema. PMID- 11058909 TI - Transthyretin Ile84Thr is associated with familial amyloid polyneuropathy. PMID- 11058910 TI - A novel missense mutation (H119L) identified in a Taiwan Chinese family with glycogen storage disease Ia (Von Gierke disease). PMID- 11058911 TI - A novel germline mutation, P48T, in the CDKN2A/p16 gene in a patient with pancreatic carcinoma. PMID- 11058912 TI - Genetic variants in the promoter (g983G>T) and coding region (A92T) of the human cardiotrophin-1 gene (CTF1) in patients with dilated cardiomyopathy. PMID- 11058913 TI - Wiskott Aldrich syndrome in an Israeli family: identification of a novel G40V mutation. PMID- 11058914 TI - Identification of a polymorphism (G83S) in the TWIST gene in Taiwanese. PMID- 11058915 TI - Two 3' polymorphisms in DLX5: g126427delTATC and g126249T-->C. PMID- 11058916 TI - A novel mutation K167X of the XLRS1 gene (RS1) in a Taiwanese family with X linked juvenile retinoschisis. PMID- 11058917 TI - Identification of two polymorphisms (c189G>C; c190T>C) in exon 2 of the human MRP6 gene (ABCC6) by screening of Pseudoxanthoma elasticum patients: possible sequence correction? PMID- 11058918 TI - Identification of a novel nonsense mutation (Q24X) in the glucose-6-phosphatase gene of a Portuguese patient with GSD Ia (von Gierke disease). PMID- 11058919 TI - Identification of a novel mutation C144F in the Notch3 gene in an Australian CADASIL pedigree. PMID- 11058920 TI - A novel SSCP variant (c.828G>A) within the M2 domain of the human neuronal nicotinic acetylcholine receptor alpha 4 subunit gene, CHRNA4. PMID- 11058921 TI - Identification of 6 new polymorphisms, g.11177G>A, g.14622C>T (R49C), g.17540T>C, g.17639T>C, g.30929T>C, g.31074G>A (R454Q), in the human microsomal epoxide hydrolase gene (EPHX1) in a French population. PMID- 11058922 TI - A novel polymorphism (g722G>A) in exon 2 of the AVPR2 gene. PMID- 11058923 TI - Management of chronic aortic regurgitation. From physician to surgeon. PMID- 11058924 TI - Myocardial revascularization with coronary endarterectomy. Stratification of risk factors for early mortality. AB - OBJECTIVE: To determine the risk factors for mortality related to myocardial revascularization when performed in association with coronary endarterectomy. METHODS: We assessed retrospectively 353 patients who underwent 373 coronary endarterectomies between January '89 and November '98, representing 3.73% of the myocardial revascularizations in this period of time. The arteries involved were as follows: right coronary artery in 218 patients (58.45%); left anterior descending in 102 patients (27.35%); circumflex artery in 39 patients (10.46%); and diagonal artery in 14 patients (3.74%). We used 320 (85.79%) venous grafts and 53 (14.21%) arterial grafts. RESULTS: In-hospital mortality among our patients was 9.3% as compared with 5.7% in patients with myocardial revascularizations without endarterectomy (p=0.003). Cause of death was related to acute myocardial infarction in 18 (54.55%) patients. The most significant risk factors for mortality identified were as follows: diabetes mellitus (p=0.001; odds ratio =7.168), left main disease (<0.001; 9.283), female sex (0.01; 3.111), acute myocardial infarction (0.02; 3.546), ejection fraction <35% (<0.001; 5.89), and previous myocardial revascularization (<0.001; 4.295). CONCLUSION: Coronary endarterectomy is related to higher mortality, and the risk factors involved are important elements of a poor outcome. PMID- 11058925 TI - Response of blood pressure to maximum exercise in hypertensive patients under different therapeutic programs. AB - OBJECTIVE: To evaluate the behavior of blood pressure during exercise in patients with hypertension controlled by frontline antihypertension drugs. METHODS: From 979ergometric tests we retrospectively selected 49 hypertensive patients (19 males). The age was 53+/-12 years old and normal range rest arterial pressure (< or = 140/90 mmHg) all on pharmacological monotherapy. There were 12 on beta blockers; 14 on calcium antagonists, 13 on diuretics and 10 on angiotensin converting enzyme inhibitor. Abnormal exercise behavior of blood pressure was diagnosed if anyone of the following criteria was detected: peak systolic pressure above 220 mmHg, raising of systolic pressure > or = 10 mmHg/MET; or increase of diastolic pressure greater than 15 mmHg. RESULTS: Physiologic response of arterial blood pressure occurred in 50% of patients on beta blockers, the best one (p<0.05), in 36% and 31% on calcium antagonists and on diuretics, respectively, and in 20% on angiotensin converting enzyme inhibitor, the later the least one (p<0.05). CONCLUSION: Beta-blockers were more effective than calcium antagonists, diuretics and angiotensin-converting enzyme inhibitors in controlling blood pressure during exercise, and angiotensin converting enzyme inhibitors the least effective drugs. PMID- 11058926 TI - PANDORA - Survey of Brazilian cardiologists about cholesterol reduction. AB - OBJECTIVE: To report about a group of physicians' understanding of the recommendations of the II Brazilian Guidelines Conference on Dyslipidemias, and about the state of the art of primary and secondary prevention of atherosclerosis. METHODS: Through the use of a questionnaire on dyslipidemia, atherosclerosis prevention, and recommendations for lipid targets established by the II Brazilian Guidelines Conference on Dyslipidemias, 746 physicians, 98% cardiologists, were evaluated. RESULTS: Eighty-seven percent of the respondents stated that the treatment of dyslipidemia changes the natural history of coronary disease. Although most of the participants followed the total cholesterol recommendations (<200mg/dL for atherosclerosis prevention), only 55.8% would adopt the target of LDL-C <100 mg/dL for secondary prevention. Between 30.5 and 36.7% answered, in different questions, that the recommended level for HDL-C should be <35mg/dL. Only 32.7% would treat their patients indefinitely with lipid lowering drugs. If the drug treatment did not reach the proposed target, only 35.5% would increase the dosage, and 29.4% would change the medication. Participants did not know the targets proposed for diabetics. CONCLUSION: Although the participating physicians valued the role played by lipids in the prevention of atherosclerosis, serious deficiencies exist in their knowledge of the recommendations given during the II Brazilian Guidelines Conference on Dyslipidemias. PMID- 11058927 TI - Clinical and pathological assessment of 82 patients with cardiovascular diseases undergoing autopsy at the Hospital das Clinicas of the Faculdade de Medicina de Botucatu from 1988 to 1993. AB - OBJECTIVE: To evaluated the clinical diagnostic, efficiency for basic death causes in patients dying of circulatory disease and de relative frequency of those diseases. METHODS: Analysis of medical record data of 82 patients, ages from 16 to 84 years old (68 over 40 years old), whose died of circulatory disease and had undergone necropsy in the period from 1988 to 1993 years in the University Hospital of Medicine Faculty of Botucatu-UNESP, Br. RESULTS: The functional class of patients were III or IV, in 78%, and 81.7% needed urgent hospitalization. By the clinical judgment the death were by ischemic heart disease in 32 (21 acute myocardial infarction), Chagas'disease in 12, valvopathy in 11, cardiomyopathy in 7, heart failure with no specification of cardiopathy in 11 and other causes in 9. At the necropsy the death cause was ischemic heart disease in 34 patients, valvopathy in 10, Chagas'disease in 10, cardiomyopathy in 5, and heart failure with no specification of cardiopathy in 2. The concordance taxes were in the same order: 94.6%, 90.0%, 83.3%, 71.4% and 28.5%. CONCLUSION: There was a great efficiency of clinical diagnosis for death cause in a general university hospital. The ischemic heart disease were the main causes of death. PMID- 11058928 TI - Outcome of acute renal failure associated with cardiac surgery in infants. AB - OBJECTIVE: To analyze the impact of acute renal failure (ARF) on the evolution of infants undergoing cardiac surgery. METHODS: We assessed 15 infants undergoing cardiac surgery who developed (ARF). Their demographic, clinical and surgical data, and evolution were analyzed. RESULTS: Their mean age was 4.4+/-4.0 months (8 days to 24 months). Twelve infants were males, and 4 patients already had ARF at surgery. The primary cause of ARF was immediate acute cardiac dysfunction in 10 infants, cardiac dysfunction associated with sepsis in 2 infants, and isolated sepsis in 3 infants. All children depended on mechanical ventilation during their postoperative period, 14 infants used vasoactive drugs, and 11 had an infectious process associated with ARF. Thirteen infants required dialytic treatment. Eleven infants developed oluguric ARF, and all had to undergo peritoneal dialysis; of the 4 patients with non-oliguric, 2 required dialysis, the main indication being hypervolemia. Of these 13 dialyzed infants, 4 died in the first 24 hours because of the severity of the underlying cardiac disease (mean urea level of 49+/-20 mg/dl). The mortality rate for the entire group was 60%, and it was higher among the patients with oliguria ARF (73% vs 25%, p<0. 001). The cause of death was acute cardiac dysfunction in 6 infants (early type-1 ARF) and sepsis in the 3 remaining infants (late type-2 ARF). CONCLUSION: The mortality rate of ARF associated with cardiac surgery in infants was hight, being higher among children with oliguria; peritoneal dialysis was indicated due to clinically uncontrolled hypervolemia and not to the uremic hypercatabolic state. PMID- 11058929 TI - Cantrell syndrome. Case report of an adult. AB - Cantrell syndrome is characterized by defects that involve the diaphragm, abdominal wall, pericardium, heart, and lower region of the sternum. It is a rare entity, usually diagnosed at birth and accompanied by high mortality due to the complexity and gravity of the anomalies. In this report, we present a 32-year-old male patient, who was diagnosed in infancy but who reached adult age asymptomatic. PMID- 11058930 TI - A 73-year-old woman with retrosternal pain but no obstructive coronary artery lesion on coronary angiography. PMID- 11058931 TI - Beta-adrenergic blocking agents in heart failure. AB - Cardiac dysfunction in heart failure is widely recognized as a progressive process, regardless of the clinical signs and symptoms. An increase in cardiac sympathetic drive is one of the earliest neurohormonal responses occurring in patients with heart failure and may be one of the major causes of the progressive remodeling leading to the decline in myocardial function, and responsible for the poor prognosis of patients with heart failure. Therefore, recent data provided by several appropriately designed clinical trials clearly indicate the benefits of beta-adrenoceptor blocking agents, combined with diuretics, ACE inhibitors, and digoxin in chronic heart failure class II to IV due to systolic ventricular dysfunction. The benefits are related to symptoms, functional capacity, remodeling, and improvement in left ventricular function, reduction in cardiovascular hospitalization, a decrease in the overall and sudden cardiac death rate, and are similar in patients with ischemic or nonischemic cardiomyopathy, independent of age, gender, or functional class. In this review we describe the cardiovascular effects of the increase in sympathetic drive, the pharmacological properties of the beta-blockers most evaluated in heart failure therapy (metoprolol, bisoprolol, and carvedilol), the major clinical trials related to these agents in heart failure, the recommendations for their appropriate use in clinical practice, the precautions to be adopted, and how to handle the more common adverse reactions. PMID- 11058932 TI - Inoculation of BALB/c mice with Lacazia loboi. AB - In a previous study, the authors inoculated Swiss mice with Lacazia loboi (L. loboi) and succeeded in maintaining a granulomatous infiltrate and viable fungal cells up to one year and six months after inoculation. Considering the experimental work on paracoccidioidomycosis, 0.03 ml of a fungal suspension obtained from a biopsy of a Jorge Lobo's Disease patient were inoculated into both hind foot pads of 32 six week-old BALB/c mice of both sexes. The animals were sacrificed 1, 4, 7 and 10 months post inoculation. The suspension contained 1.3 x 10(6) fungi/ml and presented 38% viability. Seven months after inoculation, most of the animals presented profuse infiltrates consisting of isolated histiocytes, foreign body and Langhans' giant cells and a large number of fungi, most of them viable. Emergence of macroscopic lesions was observed during the 8th month. Based on fungal count, viability index before and after inoculation, presence of macroscopic lesions and histopathological findings similar to the findings in humans, the authors believe that BALB/c mice may be a good experimental model to study Jorge Lobo's Disease, mainly regarding therapeutic evaluation. PMID- 11058933 TI - Paragonimosis in the Cajabamba and Condebamba districts, Cajamarca, Peru. AB - Stool samples from 409 pre-school and school students, living in six villages of the Cajabamba and Condebamba districts, Cajamarca, Peru, were examined using wet preparations and Lumbreras' method, looking for Paragonimus eggs. Fecal and sputum samples from two children (0. 5%) of 6 and 8 year-old showed eggs of Paragonimus. One hundred and twenty freshwater crabs, Hypolobocera chilensis eigenmanni, were collected from the Condebamba valley and 21 (17.5%) of them were infected with P. mexicanus (syn. P. peruvianus) metacercariae. Our results show the persistence of Paragonimus in human beings and in the main source of infection, the crabs. PMID- 11058934 TI - Arbitrarily primed PCR fingerprinting of RNA and DNA in Entamoeba histolytica. AB - Differences were detected in the gene expression of strains of E. histolytica using RNA (RAP-PCR) and DNA fingerprinting (RAPD). Analysis of the electrophoretic profiles of the gels revealed some polymorphic markers that could be used in the individual characterization of the strains. The 260 bands generated by using five different primers for RAP-PCR, as well as RAPD, were employed in the construction of dendograms. The dendogram obtained based on the RAPD products permitted the distinction of symptomatic and asymptomatic isolates, as well the correlation between the polymorphism exhibited and the virulence of the strains. The dendogram obtained for the RAP-PCR products did not show a correlation with the virulence of the strains but revealed a high degree of intraspecific transcriptional variability that could be related to other biological features, whether or not these are involved in the pathogenesis of amebiasis. PMID- 11058935 TI - Characterization and optimization of bovine Echinococcus granulosus cyst fluid to be used in immunodiagnosis of hydatid disease by ELISA. AB - The aim of this work was to assess the influence in the diagnostic value for human hydatid disease of the composition of bovine hydatid cyst fluid (BHCF) obtained from fertile (FC) and non-fertile cysts (NFC). Eight batches from FC and 5 from NFC were prepared and analysed with respect to chemical composition: total protein, host-derived protein, carbohydrate and lipid contents. No differences were observed in the first two parameters but carbohydrate and lipid contents were shown to be higher in batches from FC than in those from NFC. Bands of 38 and 116 kD in SDS-PAGE profiles were observed to be present in BHCF from FC only. Two pools were prepared from BHCF batches obtained from FC (PFC) and NFC (PNFC), respectively. Antigen recognition patterns were analysed by immunoblot. Physicochemical conditions for adsorption of antigens to the polystyrene surface (ELISA plates) were optimized. The diagnostic value of both types of BHCF as well as the diagnostic relevance of oxidation of their carbohydrate moieties with periodate were assessed by ELISA using 42 serum samples from hydatid patients, 41 from patients with other disorders, and 15 from healthy donors. Reactivity of all sera against native antigen were tested with and without free phosphorylcholine. The best diagnostic efficiency was observed using BHCF from periodate-treated PFC using glycine buffer with strong ionic strength to coat ELISA plates. PMID- 11058936 TI - Diagnosis of hepatitis C virus in Brazilian blood donors using a reverse transcriptase nested polymerase chain reaction: comparison with enzyme immunoassay and recombinant protein immunoblot assay. AB - Screening blood donations for anti-HCV antibodies and alanine aminotransferase (ALT) serum levels generally prevents the transmission of hepatitis C virus (HCV) by transfusion. The aim of the present study was to evaluate the efficiency of the enzyme immunoassay (EIA) screening policy in identifying potentially infectious blood donors capable to transmit hepatitis C through blood transfusion. We have used a reverse transcriptase (RT)-nested polymerase chain reaction (PCR) to investigate the presence of HCV-RNA in blood donors. The prevalence of HCV-RNA positive individuals was compared with the recombinant immunoblot assay (RIBA-2) results in order to assess the usefulness of both tests as confirmatory assays. Both tests results were also compared with the EIA-2 OD/C ratio (optical densities of the samples divided by the cut off value). ALT results were expressed as the ALT quotient (qALT), calculated dividing the ALT value of the samples by the maximum normal value (53UI/l) for the method. Donors (n=178) were divided into five groups according to their EIA anti-HCV status and qALT: group A (EIA> or = 3, ALT<1), group B (EIA> or = 3, ALT>1), grou (1< or = EIA< 3, ALT<1), group D (1< or = EIA<3, ALT>1) and group E (EIA< or = 0.7). HCV sequences were detected by RT-nested PCR, using primers for the most conserved region of viral genome. RIBA-2 was applied to the same samples. In group A (n=6), all samples were positive by RT-nested PCR and RIBA-2. Among 124 samples in group B, 120 (96.8%) were RIBA-2 positive and 4 (3.2%) were RIBA-2 indeterminate but were seropositive for antigen c22.3. In group B, 109 (87.9%) of the RIBA-2 positive samples were also RT-nested PCR positive, as well as were all RIBA-2 indeterminate samples. In group C, all samples (n=9) were RT-nested PCR negative: 4 (44.4%) were also RIBA-2 negative, 4 (44.4%) were RIBA-2 positive and 1 (11.1%) was RIBA-2 indeterminate. HCV-RNA was detected by RT-nested PCR in 3 (37.5%) out of 8 samples in group D. Only one of them was also RIBA-2 positive, all the others were RIBA-2 indeterminate. All of the group E samples (controls) were RT- nested PCR and RIBA-2 negative. Our study suggests a strong relation between anti HCV EIA-2 ratio > or = 3 and detectable HCV-RNA by RT-nested PCR. We have also noted that blood donors with RIBA-2 indeterminate presented a high degree of detectable HCV-RNA using RT-nested PCR (75%), especially when the c22.3 band was detected. PMID- 11058937 TI - An initial examination of the epidemiology of malaria in the state of Roraima, in the Brazilian Amazon basin. AB - This study firstly describes the epidemiology of malaria in Roraima, Amazon Basin in Brazil, in the years from 1991 to 1993: the predominance of plasmodium species, distribution of the blood slides examined, the malaria risk and seasonality; and secondly investigates whether population growth from 1962 to 1993 was associated with increasing risk of malaria. Frequency of malaria varied significantly by municipality. Marginally more malaria cases were reported during the dry season (from October to April), even after controlling for by year and municipality. Vivax was the predominant type in all municipalities but the ratio of plasmodium types varied between municipalities. No direct association between population growth and increasing risk of malaria from 1962 to 1993 was detected. Malaria in Roraima is of the "frontier" epidemiological type with high epidemic potential. PMID- 11058938 TI - Biotyping, serotyping and ribotyping as epidemiological tools in the evaluation of Acinetobacter baumannii dissemination in hospital units, Sorocaba, Sao Paulo, Brazil. AB - Dissemination of Acinetobacter baumannii strains in different units of a hospital in Sorocaba, Sao Paulo, Brazil was evaluated over a period of two years. By using biotyping, serotyping and ribotyping, 27 distinct clones were differentiated among 76 strains isolated between 1993-94, from clinical specimens of hospitalized patients. Two clones, 2:O4:A (biotype:serotype:ribotype) and 2:O29:A accounted for the majority of strains widely disseminated in the units during 1993. The introduction in the hospital setting, of a new clone, 6:O13:B, at the end of 1993 and its predominance through 1994 is discussed. Among 15 strains isolated from neonates, 6 (40%) belonged to the same clone, 2:O4:A. Interestingly, this clone was almost all recovered in neonatal intensive care unit, nursery and in pediatric unit. All strains were susceptible to imipenem and polymyxcin B. Multiresistant strains (up to 12 antimicrobial agents) accounted for 66.7% and 84.8% of the strains isolated in 1993 and in 1994, respectively. PMID- 11058939 TI - Envenomation by the neotropical colubrid Boiruna maculata (Boulenger, 1896): a case report. AB - This is a case report of a Boiruna maculata snake bite in a child admitted to the Hospital Municipal de Pronto Socorro de Porto Alegre, Porto Alegre, RS, Brazil. The patient was bitten on the lower left limb, and exhibited pronounced local manifestations of envenomation. She was treated with Bothrops antivenom and was discharged from the hospital five days later with marked improvement of envenomation. PMID- 11058940 TI - Massive ocular hemorrhage resulting in blindness in a patient with the sickle cell trait who developed leptospirosis. Case report. AB - This case report describes the findings of a 18 year-old black male from Bahia, a Northeastern state in Brazil, with the sickle cell trait, who developed bilateral hyphema and vitreous hemorrhage with blindness in the course of leptospirosis. The patient started to complain of blurred vision four days after the start of fever and muscular pain and approximately twelve hours after the introduction of penicillin. The severity of the leptospirosis in conjunction with sickle cell trait was considered to be the most likely explanation for this ocular complication. PMID- 11058941 TI - Susceptibility of Biomphalaria tenagophila and Biomphalaria straminea to Schistosoma mansoni infection detected by low stringency polymerase chain reaction. AB - In order to determine Schistosoma mansoni infection rates in Biomphalaria tenagophila and B. straminea, low stringency polymerase chain reaction (LS-PCR) technique was used as a complementary method to light exposure technique. LS-PCR has already been standardized in our laboratory to detect the trematode DNA in B. glabrata. Higher S. mansoni infection rates were detected using conventional method and LS-PCR. The parasite DNA profile was detected in both species after 7 day exposure to miracidia, using LS-PCR. This technique enables early detection of schistosomiasis transmission focuses, in endemic areas, before the beginning of cercariae shedding. PMID- 11058942 TI - Preliminary studies on antigenic mimicry of Ascaris lumbricoides. PMID- 11058943 TI - Aloantibodies ABO in patients with ascariasis. PMID- 11058944 TI - [Reevaluation of the nomenclature and diagnostic criteria in 477 patients with severe hepatitis]. AB - OBJECTIVE: To explore the possibility of establishing more reasonable nomenclatures and diagnostic criteria for patients with severe hepatitis (SH) through analyzing clinical data of 477 cases of SH. METHODS: The clinical characteristics and outcomes of SH were analyzed according to different criteria. RESULTS: Chronic severe hepatitis (CSH) made up 88.5% of total cases of SH. The survival rate in the patients with hepatic encephalopathy was much lower than that without hepatic encephalopathy. About 1/5 of cases of subacute severe hepatitis (SSH) and CSH had neither ascites nor hepatic encephalopathy. When the period of 2 weeks was used in replace of 10 days for the diagnosis for acute severe hepatitis (ASH), the newly added cases were consistent with the characteristics of ASH. CONCLUSION: We suggest dividing SH into 2 types: encephalopathy and non-encephalopathy by using the nomenclature of fulminant hepatitis and severe type hepatitis, respectively. The late-onset form should be added besides of acute form and subacute form. It seems to use the period of 2 weeks as the new definition of onset time for ASH. The criteria of dividing SH into 3 forms, i.e. ascites, encephalopathy and ascites plus encephalopathy, and the nomenclature recommended by the International Association for the Study of the Liver Subcommittee are not satisfactory when used in clinical cases. The typing of CSH remains to be clarified. PMID- 11058945 TI - [Prognostic evaluation of heart-liver ratio of (99m)Tc-MIBI on liver cirrhosis]. AB - OBJECTIVE: To evaluate heart-liver ratio (H/L) of (99m)Tc-MIBI on the prognosis of cirrhotic patients. METHODS: Retrospective cohort study was used to analyze the survival rate among the patients who had received H/L detection, and the H/L value was classified by Child-Pugh class. RESULTS: All the patients were followed up for 0.1 to 63 months. H/L ratio was significantly different among Child-Pugh A, B, C groups (0.85 +/- 0.15, 0.99 +/- 0.19, 1.15 +/- 0.27, F=5.09, P<0.01); 6 month, 1-year, and 2-year survival rates in H/L<1.0 group were significantly higher than those in H/L>1.0 group (85.9%, 57.4%, 57. 4% vs 50.0%, 39.4%, 19.7%, P<0.01 ). Among patients with Child-Pugh degree C, 6-month, 1-year, and 2-year survival rates in H/L<1.0 group were significantly higher than those in H/L>1.0 group (71.4%, 42.9%, 42.9% vs 27.3%, 18.2%, 0). CONCLUSION: H/L is an effective index for evaluating prognosis of cirrhotic patients, and it could be complemented by Child-Pugh classification. PMID- 11058946 TI - [Study on hepatitis C virus infection of peripheral blood mononuclear cells in chronic hepatitis C patients]. AB - OBJECTIVE: To demonstrate HCV infection and replication in the peripheral blood mononuclear cells (PBMCs) of hepatitis patients by the methods of molecular biology, immunology and electron microscopy. METHODS: HCV RNA and antigens were detected in the PBMCs of 28 patients with chronic hepatitis C by RT-PCR, Immunohistochemistry and electron microscopy, respectively. RESULTS: The positive rate of HCV RNA and HCV antigens were 77.27% (17/22) and 75.00% (21/28), respectively. Two types of spherical HCV-like particles with diameter of approximately 65 nm and 110 nm were observed in cytoplasmic vesicles by electron microscopy in the PBMCs with high titer of HCV RNA and antigens in 10 patients. The budding and shedding of these particles could also be found in the cytoplasmic vesicles. Immunoelectron microscopy using antibodies against HCV core and NS(3) demonstrated that the particles contained HCV antigens. CONCLUSION: HCV can infect and replicate in the PBMCs of patients with chronic hepatitis C. PMID- 11058947 TI - [Lymphocyte subset and its apoptosis in chronic hepatitis C]. AB - OBJECTIVE: To explore the autoimmune pathogenesis induced by hepatitis C virus. METHODS: Serum anti-GOR and peripheral blood lymphocyte subset and its apoptosis rate after activated by Pokeweed mitogen (pwm) in vitro were measured by enzyme linked immunosorbent assay and flow cytometry, respectively. Eighteen individuals with chronic hepatitis C and 16 healthy controls were observed. RESULTS: The positive rates of serum anti-GOR (66.7%) and ANA (44.4%) were significantly higher in chronic hepatitis C than normal controls (chi(2)=26.86, chi(2)=9.30, P<0.005). The apoptosis percentage of PBMCs and CD3(+)T cell was elevated in individuals with chronic hepatitis C when compared with normal controls (t=2.44, P<0.05). However, the apoptosis percentage of CD4(+)T and CD19(+)B cells in PBMCs was significantly decreased in patients with anti-GOR positive compared to anti GOR negative (t=3.17, P<0.01; t=2.294, P<0.05). Similarly, the percentage of CD4(+)CD8(+) T cells in anti-GOR positive patients was increased and CD3(+)/CD4(+) was decreased (t=2. 18, P<0.05) compared to anti-GOR negative (t=2.44, P<0.05; t=2.18, P<0.05). CONCLUSION: autoimmune response exists in chronic hepatitis C patients. There is an imbalance of lymphocyte subset and its apoptosis, which may be one of the important reasons for the mechanism of autoimmune pathogenesis of chronic hepatitis C. PMID- 11058948 TI - [The change of serum melatonin and diagnosis value in hepatic encephalopathy]. AB - OBJECTIVE: To explore the change of serum melatonin (MT) and its diagnosis value in hepatic encephalopathy (HE). METHODS: The change of MT in serum in cirrhotic patients with HE and without HE was determined by ELISA, and normal serum served as control. The change of serum MT during exacerbation and remission stage of HE was also determined. RESULTS: The level of MT in patients with HE was higher than that without (P<0.01). Both groups were higher than normal group (312.7 +/- 77.4) ng/L, (149.8 +/- 38.4) ng/L, and (77.3 +/- 28.4)ng/L, respectively, P<0.01. The level of serum MT was higher in exacerbation stage than remission stage, (308.5 +/- 59.1) ng/L and (147.8 +/- 23.3) ng/L, respectively, P<0.01. CONCLUSION: The elevation of MT in serum may be one of pathogenic factors for HE. Dynamic measurement of MT in serum is conducive to the diagnosis of HE. PMID- 11058949 TI - [Expression of collagen XVIII mRNA in rat liver fibrosis]. AB - OBJECTIVE: To measure and study quantitatively the collagen XVIII mRNA in normal and fibrotic rat livers. METHODS: We used ribonuclease protection assay to investigate the collagen XVIII mRNA expression in rat liver fibrosis induced by complete bile duct occlusion (BDO). The expression level of procollagen 1 (XVIII) mRNA was compared with that of procollagen 1 (I) and tissue inhibitor of metalloproteinase 1 (TIMP1). RESULTS: mRNA levels of procollagen and TIMP 1 increased 20- and 4-fold in BDO rat livers, respectively. In contrast, hepatic procollagen 1 mRNA level increased only 1.8-fold in fibrotic rat livers. CONCLUSION: C XVIII mRNA is upregulated slightly in liver fibrosis, which is probably correlated with the fact that CXVIII is mainly expressed by hepatocytes. PMID- 11058950 TI - [Effects of hypoxia on expression and activity of matrix metalloproteinase-2 in hepatic stellate cell]. AB - OBJECTIVE: To study the effect of hypoxia on the expression and activity of matrix metalloproteinase-2 (MMP-2) in the hepatic stellate cell (HSC). METHODS: The expression of MMP-2 and tissue inhibitor of metalloproteinase-2 (TIMP-2) in cultured rat HSC was detected by immunocytochemistry (ICC). The relative amount of MMP-2 and TIMP-2 in the culture supernatant was assayed by ELISA and the activity of MMP-2 in supernatant was examined by zymography. RESULTS: The expression of MMP-2 was enhanced in hypoxia group as compared with that in control one (t' 3.4685, P<0.05 ), while the expression of TIMP-2 was decreased (t'=2.5673, P<0.05). The activity of MMP-2 was lower in hypoxia group than in control one (t'=4.270, P<0.01). Comparing the varied duration of hypoxia, the relative amount of MMP-2 protein was the highest in 6 h group (6h vs 12h, t=4. 0945, P<0.001; 6h vs 24h, t=2.4652, P<0.025), with a contrary curve in TIMP-2. CONCLUSION: Hypoxia promotes the expression of MMP-2 and inhibits the expression of TIMP-2 in HSC. It is more notable at early stage of hypoxia. PMID- 11058951 TI - [Regulation of transcription factors c-myb and liver activator protein on expression of alpha1(I) collagen gene in activated hepatic stellate cells]. AB - OBJECTIVE: To elucidate the role of two transcription factors, c-myb and liver activator protein (LAP, a member of the C/EBP family) in the expression of alpha1(I) collagen gene in activated hepatic stellate cells (HSCs). METHODS: Rat HSCs were prepared from SD rats by in situ perfusion and single-step density Nycodenz gradient. Two chimeric luciferase reporter gene plasmids containing the human collagen alpha1(I) gene promoter fragments (-804 approximately +1452 or 804 approximately +222) were constructed. Culture-activated HSCs were co transfected with the reporter gene constructs and mammalian vectors expressing c myb or/and LAP using the cationic-liposome mediated method. RESULTS: Transient transfection of the vector expressing LAP significantly increased basal transcription from PGL(3)-col and PGL(3)-col (intron) reporter gene vectors [(587 +/- 62)U/mg protein vs (315 +/- 45)U/mg protein and (326 +/- 52)U vs (220 +/- 70)U, t=10.4 and 3.6, respectively, both P<0.05]. C-myb showed no transactivation to both PGL(3)-col and PGL(3)-col (intron) reporter gene vectors. But co expression of LAP and c-myb increased basal transcription from PGL(3)-col reporter gene by approximate 3 fold (1261 +/- 130)U vs (315 +/- 45)U, t=20.6, P<0.01. Moreover, co-expression of a specific c-myb antisense vector with the LAP vector inhibited the transactivation of LAP to collagen alpha1(I) gene promoter activity (334 +/- 29)U vs (315 +/- 45)U, t=1.06, P>0.05), which was observed only in PGL(3)-col plasmids. CONCLUSION: The transcription factor LAP transactivates collagen alpha1(I) gene in activated HSCs. C-myb plays an important role in transcriptional regulation of alpha1(I) collagen gene in HSCs and this effect is mediated by the transcription factor LAP and the cis-acting element in the first intron of alpha1(I) collagen gene. PMID- 11058952 TI - Gene therapy for murine liver cancer by adenovirus-mediated cytosine deaminase gene. AB - OBJECTIVE: To investigate anti-tumor effects of AdCD/5FC system on mice bearing MM45T.Li liver cancer. METHODS: IC(50), bystander effect and cell apoptosis were observed in vitro. After treatment of the tumor with AdCD in situ and injection of 5FC peritoneally into the mice, the anti-tumor effects were observed. RESULTS: When MOI=100, AdCD infected MM45T.Li was sensitive to 5FC, and the IC(50) was less than 50 micromol/L. When 10% MM45T.Li cells were infected with AdCD, 72% cells died after 5FC treatment, which showed significant bystander effect. Through TUNEL test and Hoechst 33258 staining, it was demonstrated that there was cell apoptosis after AdCD/5FC treatment of the MM45T.Li cell 21 day after treatment of the mice bearing tumor. The tumor volume was reduced significantly (F=20.33, P<0.05), and the median survival time was prolonged from 41-43 day to 49 day. CONCLUSION: AdCD/5FC system shows significant anti-tumor effects. PMID- 11058953 TI - Fas ligand expression and apoptosis in primary rat hepatocytes induced by lipopolysaccharide. AB - OBJECTIVE: To study hepatocyte apoptosis induced by lipopolysaccharide (LPS) directly and indirectly, and to elucidate the mechanisms of liver damage in endotoxemia. METHODS: Rat hepatocytes were isolated using collagenase perfusion, and cultured in RPMI 1640 medium. After 24h or 48 h of LPS treatment at various concentrations (1, 5, 10 mug/ml), membrane-bound Fas ligand (mFasL) expression in hepatocytes was determined by immunocytochemistry, and apoptosis was detected by TUNEL. In another set of experiments it was examined whether LPS-treated hepatocytes and its supernatants can stimulate apoptosis in LPS-untreated hepatocytes. RESULTS: LPS markedly stimulated mFasL expression and apoptosis in hepatocytes in a dose and time (24-48 h) dependent manner. In the co-culture system LPS-treated hepatocytes significantly induced LPS-untreated hepatocyte apoptosis. In contrast, there was no apoptotic cells observed in the supernatant stimulation system. CONCLUSION: LPS not only directly causes hepatocyte apoptosis, but also indirectly induces apoptosis of LPS-untreated hepatocytes by way of stimulating mFasL expression in hepatocytes. PMID- 11058954 TI - Specific regulatory inhibition of transfected HepG(2) with HCV 5'NCR by antisense phosphorothioate oligodeoxynucleotides. AB - OBJECTIVE: To screen efficient and specific new drugs against hepatitis C virus (HCV) and find the best target sequence of antisense oligodeoxynucleotides (ASODNs) targeting at HCV gene. METHODS: Fifteen S-ASODNs targeting at the 5'NCR and start AUG of HCV RNA were designed and synthesized according to the predicted RNA secondary structure of the HCV 5'NCR and the adjacent AUG region. HepG(2) cells were co-transfected with pHCV-neo4 and S-ASODN. The inhibitory effects of S ASODNs on gene expression controlled by HCV 5'NCR were determined by the assay of luciferase activity. RESULTS: Five S-ASODNs, i.e. HCV65, HCV279, HCV363, HCV349 and HCV352, showed sequence-specific and dose-dependent inhibitory activities with an inhibition rate of more than 80% at a concentration of 100 nmol/L. According to 50% inhibitory concentrations, the inhibitory activity of HCV363 was the best. On the other hand, ASODNs had only little non-specific effect by negative control trials. In addition, the results also indicated that there were some coordinate effects between ASODNs at different targets. CONCLUSION: Stem loop 2a and 3d of HCV 5'NCR and the start AUG of polyprotein precursor are candidate targets of S-ASODNs. PMID- 11058955 TI - Hepatotropism of nonenveloped DNA virus in rhesus monkey infected by transfusion transmitted virus. AB - OBJECTIVE: To study wether the nonenveloped DNA virus transmitted via blood transfusion is hepatotropic. METHODS: Total DNA was extracted from tissues of 5 experimentally infected Rhesus monkeys. A dot hybridization was done with virus double DNA strand probe or single antisense strand probe. RESULTS: Both single- and double-strand probes were hybridized with DNA of the liver, spleen, stomach, small intestine and colon. The virus was conformed present in most of all the organs when double-strand probe was used. The positive was noted only in the liver and small intestine when single-strand antisense probe was used, which showed that in liver and small intestine might have replicative intermediates of the virus. CONCLUSION: It suggests that nonenveloped DNA virus replicate in the liver and small intestine, so it might be hepatotropic. PMID- 11058956 TI - Duck hepatitis B virus DNA integration induced by oxidative radicals and its transmission from parent cells to offspring ones. AB - OBJECTIVE: To investigate the role of oxidative radicals in duck hepatitis B virus DNA (DHBV DNA) integration and the transmission of integrated DHBV DNA from parent cells to their offspring cells. METHODS: TUNEL assay and Southern blot hybridization were used to detect the cellular DNA strand breaks and the incidence of new type of DHBV DNA integration into cellular DNA of LMHD(21-6) cell line after the cells treated by low dose of hydrogen peroxide (10.0 micromol/L) during its growth from one cell to 3 +/- 10(7) to 5 +/- 10(7) cells. RESULTS: The rate of cell death induced by low dose of H(2)O(2) (10.0 micromol/L) was 32.0%, but that induced by high dose (>10.0 micromol/L) was more than 50.0%. At the same time, the incidence of new DHBV DNA integration into cellular DNA induced by 10.0 micromol/L of H(2)O(2) was 50.0% (6/12) and that in control group was 8.33% (1/12). There was a significant difference between the two groups (P<0.05). Besides, the new integrated DHBV-DNA in the parent cells could be found in the DNA of their offspring cells with the same size of base pair. CONCLUSION: Oxidative radicals is one of the important inducing factors in DHBV-DNA integration into cellular DNA and the integrated DHBV-DNA is able to transmit from the parent cells to their offspring cells stably. PMID- 11058957 TI - Construction of the recombinant retrovirus vector of HBV-S gene and it's expression in eukaryotic cells. AB - OBJECTIVE: To investigate the effectiveness of recombinant retrovirus vector in gene therapy. METHODS: The retroviral vector PLXSN-S was constructed and transferred into PA317 by means of electroporation, then HepG(2), P815, and EL4 cells were infected with the pseudovirus produced from PA317, which highly expressed HBsAg. HBsAg expression was tested by RT-PCR and ELISA. RESULTS: HBsAg was expressed variously in the eukaryotic cells mentioned above. HBsAg (A value) of the cell supernatants (48 h) were 0.92, 0.09, 0.47, respectively. CONCLUSION: The vector used in this study is an effective one to carry genes of interest to target cells and it may be useful in the test for gene therapy. PMID- 11058958 TI - Effects of carbon tetrachloride-injured hepatocytes on hepatic stellate cell activation and salvianolic acid A preventive action in vitro. AB - OBJECTIVE: To investigate the effects of carbon tetrachloride (CCl(4))-injured hepatocyte on hepatic stellate cell (HSC) activation, the relation between lipid peroxidation and liver fibrosis, and the action mechanisms of salvianolic acid A (SA-A), one of water soluble components extracted from radix salvia miltiorrhizae, against HSC activation and liver fibrosis. METHODS: The hepatocytes were isolated from the normal rats, incubated with SA-A at different concentrations, respectively, and vitamin E severed as control. At the same time the cells were fumigated with CCl(4) for 24h, and ALT activities in the medium were measured. After the cell medium was changed and cultured for another 24h, the medium was collected as "hepatocyte conditioned medium" (HCM) and measured for malondiadehyde (MDA) content. Then HCM was incubated with primary cultured HSC for 48h, and HSC proliferation, type I collagen gene expression and protein production were observed. RESULTS: ALT activity and MDA content in the hepatocyte medium were increased remarkably after the cell was injured. HCM could obviously promote HSC proliferation, type I collagen mRNA expression and protein production. SA-A could markedly inhibit ALT activity and MDA content in hepatocytes, and decreased collagen gene expression and protein production. CONCLUSION: The secretion from the peroxidative injured hepatocyte could stimulate HSC activation, SA-A could decrease activation by alleviation of hepatocyte peroxidation. PMID- 11058959 TI - Effects of angiotensin II receptor blockade on hepatic fibrosis in rats. AB - OBJECTIVE: To investigate the effects of angiotensin II type 1 receptor blockade, losartan, on serum levels of components of extracellular matrix in experimental fibrotic rats. METHODS: Fifty male Spague-Dawley rats were separated into five groups (control, model, and 3 treatment groups). Excepting rats in control group, all rats were given subcutaneous injection of 40% carbon tetrachloride (once every 3 days for 6 weeks). Rats in 3 treatment groups were also given losartan of 10mg/kg, 5mg/kg, 2.5mg/kg daily for 6 weeks via gastrogavage, respectively. At the end of sixth week, all rats were sacrificed. Radioimmunoassay was performed to determine the serum levels of hyaluronic acid (HA), Laminin (LN), procollagen type III (PCIII) and collagen type IV. Van Giesion collagen staining was used to evaluate the extracellular matrix of the liver tissue. RESULTS: Compared with model group, losartan significantly reduced the serum levels of HA [from (911.66 +/- 345.49)microg/L to (425.05 +/- 115.80)microg/L], LN [from (209.87 +/- 91.57)microg/L to (83.56 +/- 22.12)microg/L, PCIII [from (31.82 +/- 6.90)microg/L to (22.78 +/- 8.38)microg/L] and collagen IV [from (54.09 +/- 19.81)microg/L to (30.51 +/- 12.39)microg/L] (P<0.05) and greatly attenuated the degree of liver fibrosis (P<0.05). CONCLUSION: Losartan can markedly reduce the serum levels of LN, HA, PCIII and collagen type IV of fibrotic rats induced by CCl(4) and greatly attenuate the degree of liver fibrosis. PMID- 11058960 TI - Rat hepatocellular apoptosis induced by glycodeoxycholate. AB - OBJECTIVE: To explore the relationship between glycodeoxycholate (GDC) and rat hepatocellular apoptosis. METHODS: GDC was used to treat rat hepatocytes cultured in vitro and the apoptotic cells were observed and analyzed with light microscope, transmission electron microscope, TUNEL in situ hybridization, DNA agarose gel electrophoresis and flow cytometry. RESULTS: When rat hepatocytes were incubated with GDC (final concentration of 100 micromol/L) for 2h, apoptotic hepatocytes were observed with light and transmission electron microscope, and TUNEL in situ hybridization. When the hepatocytes were incubated with GDC (final concentration of 50, 100, 150 micromol/L, respectively) for 2h, or with GDC (final concentration of 100 micromol/L) for 2, 4, 6, 8h, respectively, agarose gel electrophoresis of the hepatocytes DNA demonstrated the typical ladder patterns. CONCLUSION: GDC with final concentration of 50, 100, 150 micromol/L could induce rat hepatocellular apoptosis. PMID- 11058961 TI - Modulation of the far-upstream enhancer of the rat alpha-fetoprotein gene by members of the ROR alpha, Rev-erb alpha, and Rev-erb beta groups of monomeric orphan nuclear receptors. AB - Expression of the oncodevelopmental alpha-fetoprotein (AFP) gene is tightly regulated and occurs in the yolk sac, fetal liver and intestine, and cancerous liver cells. Transcription of the AFP gene is under the control of three enhancers that are very tissue specific. We have shown that the most upstream of these enhancers, located at -6 kb, works through the combined action of liver enriched factors and nuclear receptors that bind to three regions of this DNA regulatory element. This study showed that orphan nuclear receptors of the ROR alpha, Re-verb alpha, and Rev-erb beta groups can bind as monomers with high affinity and specificity to an evolutionarily conserved AGGTCA motif in the functionally important region 1 of this AFP enhancer. Transient transfection experiments performed with human HepG2 hepatoma cells showed that overproduction of ROR alpha 4 stimulated the activity of the AFP enhancer in a dose-dependent manner, while that of Rev-erb alpha and Rev-erb beta had the opposite effect. These effects were highly specific and required the integrity of the AGGTCA motif. The action of these nuclear receptors also occurred in the context of the entire 7-kb regulatory region of the rat AFP gene. These results suggest that altering the amounts or activities of these orphan receptors in cells of hepatic or endodermal origin could modulate AFP gene expression in response to a variety of developmental or carcinogenic stimuli. PMID- 11058962 TI - Genomic structure of murine mitochondrial DNA polymerase-gamma. AB - We have sequenced a genomic clone of the gene encoding the mouse mitochondrial DNA polymerase. The gene consists of 23 exons, which span approximately 13.2 kb, with exons ranging in size from 53 to 768 bp. All intron-exon boundaries conform to the GT-AG rule. By comparison with the human genomic sequence, we found remarkable conservation of the gene structure; the intron-exon borders are in almost identical locations for the 22 introns. The 5' upstream region contains approximately 300 bp of homology between the mouse and human sequences that presumably contain the promoter element. This region lacks any obvious TATA domain and is relatively GC rich, consistent with the housekeeping function of the mitochondrial DNA polymerase. Finally, within the 5' flanking region, both mouse and human genes have a region of 73 bp with high homology to the tRNA-Arg gene. PMID- 11058963 TI - Subphases of DNA replication in Drosophila cells. AB - Exponentially growing Drosophila S2 cells in suspension culture were synchronized at low- and high-resolution centrifugal elutriation, and DNA synthesis was measured by [(3)H]-thymidine incorporation throughout the S phase. At low resolution, one repair peak at the G(1)/G(0) border and two replication peaks known as early and late S subphases were observed. At high resolution, six chronologic compartments were distinguished. The distribution of these peaks indicated one repair peak at 2.05 C value, one minor replication peak at 2.43C, and four major subphases of replication corresponding to 2.64C, 2.89C, 3.32C, and 3.60C, representing 6.7%, 3.4%, 15.3%, 20.4%, 32.1%, and 22.0% of the synthetic activity, respectively. The five major peaks of cell growth with 2.32C, 2.56C, 2.85C, 3.18C, and 3.58C values consistently preceded those of replication subphases. PMID- 11058964 TI - Competition with TATA box-binding protein for binding to the TATA box implicated in human cytomegalovirus IE2-mediated transcriptional repression of cellular promoters. AB - The human cytomegalovirus IE2 is a sequence-specific DNA-binding protein. A consensus IE2-binding site (IBS) contains two copies of the dinucleotide CG separated by 10 not well-conserved but AT-rich nucleotides. In this report, we demonstrated that the TATA box of the insulin-like growth factor binding protein 4 (IGFBP4) promoter is embedded in an IBS. In a transient transfection study, IE2 mediated repression of a reporter driven either by a synthetic promoter containing the IGFBP4 TATA/IBS element or by the native IGFBP4 promoter was dependent on the intactness of the IBS. Competition with TBP for binding to the IGFBP4 TATA/IBS element may underlie the mechanism for the IE2-mediated repression, because IE2 and TATA box-binding protein (TBP) binding to the IGFBP4 TATA/IBS element are mutually elusive. Moreover, the TATA boxes of several other genes, including ADA and CPH-70, are likewise confined in IBS-like sequences. The IE2 interacts with those TATA/IBS elements in vitro and inhibits transcription driven by them in vivo, supporting the idea that competitive inhibition of TBP binding to the TATA box represents a novel mechanism exploited by IE2 to repress cellular gene expression. PMID- 11058966 TI - Casting a ballot for primary prevention. PMID- 11058965 TI - DNA sequence requirements for the activation of 434 P(RM) transcription by 434 repressor. AB - A dimer of the 434 repressor bound at O(R)2 activated transcription initiation from P(RM) by contacting RNA polymerase. Although DNA-binding site mutations at either end of O(R)2 decreased the ability of the repressor to activate P(RM) transcription, mutations proximal to the promoter had a greater effect on transcription activation. Orienting a repressor subunit bearing the altered specificity Gln-28 --> Ala mutation to the halfsite of O(R)2 proximal to the P(RM) promoter decreased the repressor's ability to activate transcription initiation at 434 P(RM) to a much greater extent than if this subunit was placed in the O(R)2 half-site distal to P(RM). In addition to showing that the downstream (promoter proximal) subunit of the O(R)2-bound 434 repressor functions in activating 434 P(RM), the results indicated that DNA sequence-dependent conformational changes alter the efficiency with which the repressor activates P(RM) transcription. These unexpected findings highlight the importance of the structure of the repressor-DNA interface in activating transcription from P(RM). PMID- 11058967 TI - Nurses should not foster enduring pain in elderly patients. PMID- 11058968 TI - Reader interested in increased paclitaxel reactions. PMID- 11058969 TI - Opioid tapering in hematopoietic progenitor cell transplant recipients. AB - PURPOSE/OBJECTIVES: To describe current opioid-tapering practice, patient pain levels and withdrawal symptoms, and nurse documentation during opioid tapers. DESIGN: Descriptive, exploratory, prospective, quantitative, and qualitative. SETTING: A 32-bed blood and marrow transplant unit in a large, tertiary U.S. care center. SAMPLE: 45 blood and marrow transplant recipients between the ages of 7 64. Types of transplant were autologous, allogeneic, and unrelated donor marrow; peripheral blood stem cell; and umbilical cord blood. METHODS: In daily interviews, patients reported pain levels and withdrawal symptoms during opioid tapers. Demographic, medication, and nurse documentation data were obtained from patient hospital records. MAIN RESEARCH VARIABLES: Taper length, daily opioid dosage, pain, withdrawal symptoms, and nurse documentation. FINDINGS: Length of taper ranged from 1-17 days (X = 6.53, SD = 4.26). Analysis of variance indicated no difference by disease or transplant type in length of taper, cumulative opioids given pretaper or during taper, or number of self-reports of withdrawal symptoms. Daily changes in nurse-administered opioid dosage during tapers ranged from a decrease of 67% to an increase of 14%. Children received significantly more opioids/kg during taper than adults. Means of adults' and children's self reports of pain were low. The means of patient-reported withdrawal symptoms were highest on taper days two through six. Nurse documentation was sparse. CONCLUSIONS: Daily taper rates vary widely, pain is well managed, and most patients experience several withdrawal symptoms. IMPLICATIONS FOR NURSING PRACTICE: Use of an opioid-taper guideline may promote consistency of tapering while not increasing levels of pain or withdrawal symptoms. PMID- 11058970 TI - Components of social support among Japanese women with breast cancer. AB - PURPOSE/OBJECTIVES: To describe social support from the perspective of Japanese women with breast cancer. DESIGN: Qualitative, descriptive, and interpretive. SETTING: Outpatient breast clinic at a cancer center in a metropolitan area in Japan. SAMPLE: Sixteen Japanese women diagnosed with stage I and II breast cancer who underwent either modified radical or radical mastectomy and were receiving outpatient follow-up care. The majority were in their 50s, married, and employed. METHODS: Narrative data were collected through individual interviews using a semistructured interview guide. Responses were analyzed using qualitative, inductive content analysis. MAIN RESEARCH VARIABLE: Social support experiences. FINDINGS: "Simply Do It!," "Doing With/Being With," and "Doing For" were three situations in which Japanese women described social support. Doing For was the provision of instrumental aid and was most consistent with Western findings. Each of the three situations was perceived as emotional support. Cultural norms of reciprocity and social obligation entailed strong social imperatives that were sources of conflict. Verbalizations of personal feelings and emotions often did not occur, even among closest family and friends, but were communicated by behavior. CONCLUSIONS: The influence of Japanese culture on interpersonal relationships is strong and guided by cultural norms that include social obligation, maintaining harmonious relationships, and reciprocity. Provision of instrumental aid and areas of conflict were most consistent with Western findings, but differences in sources and forms of social support were pronounced. Continued exploration of social support in Japanese women with breast cancer is warranted. IMPLICATIONS FOR NURSING PRACTICE: Understanding and valuing cultural diversity is vital to developing more effective and individualized nursing care consonant with patients' and families' desires and comfort. PMID- 11058971 TI - Cancer survivors and quality of life: a critical review of the literature. AB - PURPOSE/OBJECTIVE: To review and analyze the nature of quality of life (QOL) as a critical construct in psychosocial oncology research and to offer direction for future research in studies of cancer survivors. DATA SOURCES: Select and representative empirical studies of cancer survivors and QOL outcomes. DATA SYNTHESIS AND CONCLUSIONS: A consensus exists that the concept of QOL is multidimensional and subjective in nature and thereby requires a methodologic approach that examines the cognitive processes by which individuals achieve QOL. IMPLICATIONS FOR NURSING PRACTICE: Healthcare providers best ensure adequate clinical care for cancer survivors when both the biomedical and psychosocial effects of disease on health, well-being, and function are known as well as the varied ways in which these individuals adapt and live the remainder of their lives. PMID- 11058972 TI - An analysis of the readability and cultural sensitivity of information on the National Cancer Institute's Web site: CancerNet. AB - PURPOSE/OBJECTIVES: To ascertain the level of reading skills required by lay people who access patient-related cancer information through CancerNet, the Web site of the National Cancer Institute (NCI), and to assess the cultural sensitivity of the information targeted to certain ethnic groups. DESIGN: Descriptive study, repeated measures. SETTING: NCI's CancerNet Web site (http://cancernet.nci.nih.gov). SAMPLE: A convenience sample of 49 documents from the CancerNet Web site was analyzed. METHODS: The readability of each document was analyzed using the Flesch-Kincaid readability formula. Using Bloch's Ethnic/Cultural Assessment Tool as a guide, a content analysis of the ethnic related documents was performed to determine the cultural sensitivity of the information. MAIN RESEARCH VARIABLES: Readability and cultural sensitivity of the written cancer information. FINDINGS: The overall mean reading level was 12th grade. Little variation existed in the cultural content of the information even though several ethnic groups were targeted. CONCLUSION: Information on CancerNet needs to be modified to meet the information needs of people with low reading skills and to make the information more culturally sensitive for various ethnic groups. IMPLICATIONS FOR NURSING PRACTICE: Finding successful methods for educating patients and families is a primary responsibility of oncology nurses. Information can play a vital role in helping patients to engage in self-care behavior. Therefore, nurses must be knowledgeable about the readability, usefulness, and cultural-sensitivity of information on cancer Web sites so that they can guide patients to appropriate Internet resources. PMID- 11058973 TI - Knowledge and attitudes of nurses in veterans hospitals about pain management in patients with cancer. AB - PURPOSE/OBJECTIVES: To assess nurses' knowledge and attitudes about pain management and patients in pain. DESIGN: Exploratory, descriptive. SETTING: Seven medical-surgical inpatient units in two large veterans hospitals in Southwest Florida. SAMPLE: A convenience self-select sample of 85 nurses (RNs and LPNs working on the target units on all shifts). METHODS: Staff nurses were approached at work and asked to complete the data collection forms. MAIN RESEARCH VARIABLES: Knowledge about pain management principles and attitudes toward pain management and patients in pain. FINDINGS: Areas of major knowledge deficits included physiology of pain and pharmacology of analgesics. Nurses were most knowledgeable about the importance of asking patients about their pain, around-the-clock scheduling, tolerance, and use of distraction. Patient behavior, age, and gender seemed to unduly influence nurses in their pain management decisions. Regarding attitudes about pain management, the majority of nurses did not agree that patients and their families should have the most control over analgesic scheduling and that a constant level of analgesic should be maintained in the blood. In fact, 82% indicated that around-the-clock analgesics increase the risk for sedation and respiratory depression. CONCLUSIONS: Years after the Agency for Health Care Policy and Research published pain guidelines, nurses in veterans hospitals continue to lack knowledge and have negative attitudes that may negatively affect pain management in patients with cancer. IMPLICATIONS FOR NURSING PRACTICE: Basic and continuing education for nurses needs to include intensive content about pain management. Continued research is needed to document improvements in pain management by nurses. PMID- 11058974 TI - Evaluating patient distress from cancer-related fatigue: an instrument development study. AB - PURPOSE/OBJECTIVES: To develop a clinically useful and psychometrically sound tool for the measurement of the distress from fatigue in patients with cancer. DESIGN: Instrument development of a numeric rating scale to assess cancer-related fatigue (CRF) distress. SETTING: A comprehensive cancer center and a Veterans Administration hospital. SAMPLE: The interview sample consisted of 17 adults with cancer experiencing CRF. The sample for instrument testing consisted of 221 adults with cancer. Twenty-nine different types of cancers were among the diagnoses. Seventy-eight percent of the participants were receiving some form of cancer treatment. The mean age was 60. METHODS: The Cancer-Related Fatigue Distress Scale (CRFDS) was developed from 23 in-depth audiotaped interviews with patients experiencing CRF. Krippendorff's content analysis procedures were used for interview analysis. Twenty-three items were generated. Five cancer survivors assessed content validity. Factor analysis was used to assess construct validity, and coefficient alpha was used to estimate the reliability of the CRFDS. FINDINGS: Factor analysis resulted in all items loading on one factor, indicating a single scale. The coefficient alpha was 0.98 after the elimination of three items. CONCLUSIONS: The CRFDS has strong content validity, high reliability, and very good construct validity. IMPLICATIONS FOR NURSING PRACTICE: The CRFDS is a clinically useful and psychometrically sound tool for the measurement of CRF distress. This tool is clinically useful because of its brevity (20 items), clear instructions that required no training to use, and a readability score at the third-grade level. PMID- 11058975 TI - Factors that influence a patient's decision to participate in a phase I cancer clinical trial. AB - PURPOSE/OBJECTIVES: To explore factors that influence an individual's decision to participate in phase I cancer clinical trials. DESIGN: Exploratory, descriptive, and qualitative study. SETTING: Outpatient oncology clinic of a comprehensive cancer center in an urban setting. SAMPLE: 22 patients with cancer who had been asked at some point during their treatment to participate in a phase I cancer clinical trial were invited to participate in a focus group. Eight patients participated. METHODS: Two focus groups were conducted. MAIN RESEARCH VARIABLES: Factors that influenced patients' decisions to participate in a phase I cancer clinical trial. FINDINGS: Participants expressed that hope for a cure and trusting the oncologist's advice were the primary factors that influenced their decisions to participate in phase I research. The majority expressed surprise that anyone would participate in an experimental study for altruistic reasons. CONCLUSIONS: Patients choose to hope for some personal benefit from their participation in phase I clinical trials even though they realized that is not the purpose or goal of the trial. IMPLICATIONS FOR NURSING PRACTICE: Nurses are in an ideal position to offer information to individuals who are deciding whether to participate in a phase I cancer clinical trial. However, most nurses are not seizing this opportunity. Patients are unable to describe nurses' impact on their decision to enter a phase I cancer clinical trial. PMID- 11058976 TI - Correlates of fatigue during and following adjuvant breast cancer chemotherapy: a pilot study. AB - PURPOSE/OBJECTIVES: To examine patterns of and relationships between activity, sleep, symptom distress, health status, and fatigue during and following adjuvant breast cancer chemotherapy with doxorubicin and cyclophosphamide. DESIGN: Prospective, descriptive, repeated measures. SETTING: Midwestern, urban, oncology clinics. SAMPLE: 14 women, ages 32-69 (X = 52.4), with stage I or II breast cancer receiving four cycles of chemotherapy. METHODS: Wrist actigraph, modified Morin Sleep Diary, Symptom Experience Scale, Medical Outcomes Study-Short Form 36, and Piper Fatigue Scale were used to collect data two days prior to and during the 21-day cycle 3, and for three days at three weeks and two months following treatment 4. MAIN RESEARCH VARIABLES: Activity, sleep, symptom distress, health status, and fatigue. FINDINGS: Fluctuating patterns of lowered activity, disturbed sleep, mild-to-moderate symptom distress, and moderate fatigue were identified. Mean activity levels ranged from 65%-80% of norms during and following treatments. Patterns of sleep (total rest, sleep latency, wake after sleep onset, and sleep efficiency) differed from established norms. Patients experienced the highest levels of fatigue and symptom distress during the first four days after treatment 3. Correlates of fatigue were greater symptom distress, lower activity, and poorer physical and social health status; variables representing disturbed sleep trended toward associations with fatigue. CONCLUSION: Activity, sleep, symptom distress, and health status cluster in patterns associated with either lower or higher fatigue. IMPLICATIONS FOR NURSING PRACTICE: Efforts to manage symptoms, remain active, and obtain quality sleep, especially in women with poorer health status, may assist in modifying fatigue. PMID- 11058977 TI - Introduction: cancer prevention and early detection--from thought to revolution. PMID- 11058978 TI - Application of carcinogenesis theory to primary prevention. AB - PURPOSE/OBJECTIVES: To apply current cancer development theory to concepts of and interventions for cancer prevention. DATA SOURCES: Current medical and nursing literature. DATA SYNTHESIS: Carcinogenesis is a complex process that is only partially understood. The information that is available supports efforts to control the disease. However, the lack of detailed understanding limits attempts to alter the course of cancer. CONCLUSION: The understanding that cancer is the result of cumulative genetic errors can be used to direct oncology nursing interventions to prevent the disease. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses should be aware of the various factors that influence cancer development to prescribe primary prevention measures. Nurses also must be aware that efforts to understand carcinogenesis and exploit that knowledge are ongoing. Awareness of advances in the understanding of cancer development are keys to appropriate patient care. PMID- 11058979 TI - Issues in determining cancer screening recommendations: who, what, and when. AB - PURPOSE/OBJECTIVES: To provide an overview of the issues involved in evaluating and choosing cancer screening guidelines. DATA SOURCES: Current epidemiologic literature. DATA SYNTHESIS: Screening is universally accepted as being important for cancer control. A number of organizations have developed cancer screening guidelines. However, agreement on who should be screened, with what tests, and at what ages and intervals is far from universal. Choice of screening guidelines may be influenced by differences in populations of interest and considerations of screening effectiveness. CONCLUSION: Directing patients toward appropriate screening tests and intervals requires attention to information about screening tests and to individual and institutional characteristics. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses should be aware of the various issues that influence cancer screening recommendations. Nurses advising patients about or developing cancer screening services must consider identifying both the patient population to be screened and factors influencing screening test effectiveness. This information then can be used to choose the most appropriate guidelines from those available. Consideration of the issues also can be applied as advances in screening evolve. PMID- 11058980 TI - The role of the nurse in developing cancer screening programs. AB - PURPOSE/OBJECTIVE: To describe and provide an overview of the role that nurses can play in developing cancer screening programs. DATA SOURCES: Published articles, book chapters, and clinical experience. DATA SYNTHESIS: Many approaches and considerations are available for developing cancer screening programs. Planning considerations include needs assessment, patient-education strategies, funding, recruitment issues, follow-up strategies, staffing, policy development, and evaluation. An understanding of these considerations may lead to more effective cancer screening programs. CONCLUSIONS: Nurses can play an integral role in the development and management of cancer screening programs. Careful program planning may enhance this process. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses need to consider a wide range of issues when developing cancer screening programs, including the purpose of the program, how the program will be implemented, the population being served, and the potential impact on individuals being served. PMID- 11058981 TI - Cancer genetics nursing: impact of the double helix. AB - PURPOSE/OBJECTIVES: To describe the impact of genetic information on oncology nursing practice and to identify roles for oncology nurses in the field of cancer genetics. DATA SOURCES: Published articles, abstracts, books, and clinical experience. DATA SYNTHESIS: Oncology nurses in all areas of practice are affected by the recent explosion of genetic information. The identification of genetic mutations associated with increased risk for certain cancers and subsequent development of cancer predisposition testing have created a tremendous need for health care professionals who can explain and interpret genetic information. CONCLUSIONS: Oncology nurses already have many basic skills essential in the management of genetic information. An area in which oncology nurses may have the most to offer is in helping patients who carry a genetic predisposition to cancer understand the cancer surveillance and risk-management options available to them. Oncology nurses will be on the forefront in helping patients understand what this information means to them and how to apply it to their lives. IMPLICATIONS FOR NURSING PRACTICE: As we enter the 21st century, the advances in genetic information and the impact of the Human Genome Project will change oncology nursing as we know it. Oncology nurses will be expected to assess and interpret genetic and nongenetic cancer risk as an integrated whole, including genetic risk factors, environmental and lifestyle risk factors, and the interaction of the two. The challenge will be to make this complex information meaningful to patients as they make choices to manage their own cancer risk. PMID- 11058982 TI - Bioethical considerations in cancer prevention and early detection practice and research. AB - PURPOSE/OBJECTIVES: To review current ethical issues in cancer prevention and early detection, discuss some of the key methods and theories in contemporary bioethics, present clinical scenarios that illustrate moral problems that nurses may encounter, and show how the theories and methods can assist ethical decision making. DATA SOURCES: Published articles, abstracts, and books; case studies developed from clinical experience and literary sources. DATA SYNTHESIS: Oncology nurses face a host of ethical considerations when providing cancer prevention and early detection education and services and when conducting research in this arena. A variety of ethical methods and theories--including principlism, casuistry, utilitarianism, rights-based theories, virtue ethics, ethics of care, and communitarian ethics--may assist nurses in tackling ethical challenges. CONCLUSIONS: The growing field of cancer prevention and early detection raises a number of moral issues that have implications for oncology nursing. Because no single method or theory provides a complete account of bioethics, ethical problem solving should be approached using a variety of methods and theories whereby they can be coherently and appropriately applied together. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses need to increase their awareness of ethical issues related to cancer prevention and control and consider them when planning patient care and research programs. PMID- 11058983 TI - Educational programs in cancer prevention and detection: determining content and quality. AB - PURPOSE/OBJECTIVES: To provide an overview of available programs and to present recommendations for important components of a cancer prevention and detection educational and training experience. DATA SOURCES: Research, journal articles, Internet resources. DATA SYNTHESIS: Variations in the efforts to provide basic, advanced, and continuing medical education exist across healthcare disciplines. These variations suggest that differences in interest and in the perceptions of the most urgent cancer control issues may exist across disciplines. CONCLUSION: Nursing should increase its involvement in the development and testing of quality cancer prevention and detection programs that provide a multidisciplinary approach without losing the important nursing perspective. IMPLICATIONS FOR NURSING PRACTICE: The integration of this information may be helpful in enhancing nursing efforts to develop greater expertise in cancer control and may facilitate a multidisciplinary approach to cancer prevention and detection that includes essential nursing perspectives. PMID- 11058984 TI - Addressing issues for early detection and screening in ethnic populations--an update. PMID- 11058985 TI - Hysteria and the medical narrative. PMID- 11058986 TI - Tlatilco sculptures, diprosopus, and the emergence of medical illustrations. PMID- 11058987 TI - Why not call modern medicine "alternative"? PMID- 11058988 TI - Mind-body unity: Gregory of Nyssa and a surprising fourth-century ce perspective. PMID- 11058989 TI - Medieval contributions to the search for truth in clinical medicine. PMID- 11058990 TI - Charcot and Duchenne: of mentors, pupils, and colleagues. PMID- 11058991 TI - Must doctors still examine patients? PMID- 11058992 TI - Xenotransplantation in the new millennium: moratorium or cautious experimentation? PMID- 11058993 TI - Truth, justice, and genetics. PMID- 11058994 TI - Aesthetics in the swamps. PMID- 11058995 TI - Thomas Gold's deep hot biosphere and the origin of petroleum [1]. Essay review. PMID- 11058996 TI - [Prions in yeast and filamentous fungi]. PMID- 11058997 TI - [Heterodimeric nuclear receptors. I. Vitamin and hormone receptors]. PMID- 11058998 TI - [Heterodimeric nuclear receptors. II. Fatty acid and steroid metabolism regulation]. PMID- 11058999 TI - [Nitric oxide synthase genes: structure, regulation of expression, protein products]. PMID- 11059000 TI - [The group of protein kinases CKI]. PMID- 11059001 TI - [Protein kinase CKII]. PMID- 11059002 TI - [Complex I NADH-dehydrogenase of the respiratory chain. Basic subunits encoded by a nuclear genome]. PMID- 11059003 TI - [Mitochondrial genes of complex I NADH dehydrogenase respiratory chain subunits. Editing of their transcripts]. PMID- 11059004 TI - [Na+/H+ antiporters in mammalian cells]. PMID- 11059005 TI - [Sequencing of the human genome--two projects, two strategies]. PMID- 11059006 TI - [Significance of A. A. Ukhtomskii for the modern science]. PMID- 11059007 TI - [Experimental confirmation of A. A. Ukhtomskii's ideas of the dominant]. PMID- 11059008 TI - [A. A. Ukhtomskii's ideas in studies on the cognitive activity]. AB - Experimental data on spatial and temporal organisation of the human EEG from birth to maturity, at resting and during cognitive activity, is presented. The data obtained is interpreted in light of a confirmation of the Ukhtomsky's ideas on the "functional working organ" appearing in the brain for realising cognitive processes. Role of the alpha-rhythm in formation of plastic intracortical neuronal network is elucidated. A hypothesis is advanced stating that the gradually developing brain system exerting a local regulated activating effects upon the cortical level, is a significant factor of the functional intercentre integration. PMID- 11059009 TI - [Psychophysiology of set and the theory of dominant]. AB - The regulatory function of the set and dominanta in human cognitive activity is discussed. Role of the stimuli context in formation of the set is demonstrated. PMID- 11059010 TI - [Neurochemical organization of the brain reinforcing systems]. AB - Modern concepts of neurophysiological mechanisms of the brain reinforcing systems are reviewed from A.A. Ukhtomsky's concept of dominanta. The brain mesocorticolimbic system was shown to play a key role in functioning of the brain reinforcing systems. Morphological and neurochemical organisation of this system determining the emotional sphere, was studied. The dopamine system was found to be the main neurochemical tool of the mesocorticolimbic system. Other transmitters, neuromodulators, and hormones play a regulatory role in the latter. The data obtained corroborate the authors' concept of fluctuating emotional gradient. PMID- 11059011 TI - [A. A. Ukhtomskii's ideas of correlation between the temporal and the spatial factors in the nervous system activity]. PMID- 11059012 TI - [Reflection of the type and successfulness of the cognitive activity in the human EEG: application of nonlinear methods in psychophysiology]. AB - The information processing by the human brain was studied in two types of cognitive activity: solving a visual-spatial task and a verbal task. One of the Wechsler tests represented a visual-spatial task, whereas a speed-reading was used as a verbal task. A correlation dimension calculated from the EEG time series, was shown to depend on individual capabilities. Subjects with a higher correlation dimension in frontal brain areas executed more successfully the spatial task. PMID- 11059013 TI - [The principle of dominant and cortical mechanisms of the audiomotor correlation]. AB - The data obtained in behavioural, neurophysiological, and bioacoustic experiments revealed plasticity of functional organisation of the auditory input in the cat sensory-motor cortex and are reviewed from A.A. Ukhtomsky's "Dominanta" principle's standpoint. PMID- 11059014 TI - [Steroids in the reproductive function regulation in fish]. AB - Following ovulation cortisol and testosterone concentrations were lower in the stellate sturgeon coelomic fluid than in the blood serum, whereas the levels of progesterone was lower in the blood. Specific cytosol binding of sex steroids in the brain and the gonads differed in fish before and after the maturation. In the forebrain, the specific binding of androgens dropped sharply after the ovulation. In the hypothalamus the specific binding of estrogens was the highest after the final maturation and ovulation. A certain correlation between the blood sex steroids concentrations and their specific binding in the brain and gonads cytosol, were established. PMID- 11059015 TI - [Changes in the animal behavior caused by sequential changes of dominants related to reproduction]. AB - Female rats during the sex dominanta (proestrus) revealed lower anxiety scores and a higher general level of activity than the rats in diestrus. Responses to stress were also more obvious in proestrus. During gestation the rats revealed a lower activity and a higher anxiety. In lactation, the rats' behaviour was similar to that in proestrus. PMID- 11059016 TI - [Dynamics of the dominance of identified cardioregulatory neurons in the snail Achatina fulica] . AB - 9 cardioregulating neurones belonging to 5 different functional groups were studied in visceral and right parietal ganglia of the Giant African snail Achatina fulica. The neuronal network included multimodal and multifunctional cells exerting short- or long-lasting chronoionotropic effects on the cardiac electro- and mechanograms. Mechanisms of the differences in the cardioregulating effectiveness of these groups were discussed. PMID- 11059017 TI - [Dominant structural and functional adaptations of distant chemosensory systems in phylogenesis of Gastropoda]. AB - The invasion of land by gastropods independent and repeated in the course of their evolution, was shown to be accompanied by appearance of organisationally similar olfactory tentacular organs and special integrative centres. The majority of primary and secondary water gastropods in different phylogenetic groups had a different, more primitive organisation of the tentacular sensory system as compared to the terrestrial species. Regularities of the phylogenetic adaptations of mollusks to the habitat media and lifestyle are discussed. PMID- 11059018 TI - [Psychophysiological factors of changes in the functional state affecting the quality and effectiveness of human higher mental functions]. AB - A number of factors were shown to affect human functional state: a decrease in the tolerance of transitory hypoxia, an enhanced excitability in the brain stem structures, etc., with the aid of infraslow electrical processes. PMID- 11059019 TI - [Non-transmitter effect of acetylcholine in the neuromuscular preparation]. AB - Acetylcholine was shown to maintain the efficiency of a fatigued muscle of rat increasing the evoked transmitter release and inducing a hyperpolarization of the muscle fibre membrane. The effects proved to be long-term ones. Modulatory effects of acetylcholine were shown to be realized via structures which differed pharmacologically from the typical n- and m-cholinoreceptors with the participation of ouabain-sensitive isoforms of Na+, K(+)-ATPase. The data obtained corroborates existence of a long-term neuronal regulation of the neuromuscular transmission efficiency involving non-quantal acetylcholine. The regulatory pathways are supposed to be different in muscle fibres with different functional characteristics and different ability of adaptation under physiological loading. PMID- 11059020 TI - [Ca(2+)-signaling in peritoneal macrophages]. AB - Mechanisms of the Ca2+ signal generation and regulation in peritoneal macrophages activated with purinergic agonists (ATP, UTP), as well as endoplasmic Ca(2+) ATPase inhibitors, were investigated. Using a wide range of drugs affecting the intracellular signaling systems' components, an important role of second messenger systems and other key functional cellular systems in Ca2+ signals regulation in the macrophages, was shown. PMID- 11059021 TI - [Pinealocyte functioning in stress during daytime in rats]. AB - Several types of unit activity were detected in the rat pineal gland during a 48 hour water and food deprivation. The unit activity rate in stress was higher by 4 6 times than in intact rats (due to an increase in the "fast" cells number and switching of some cells from regular to a burst type of activity). Electrical stimulation of olfactory epithelium decreased the unit activity rate in the most active pinealocytes. The daytime increase in the pineal electrical activity reflects an intensification of secretion of the protein/peptide substances and, probably, serotonin but not the melatonin. Blocking exocytosis with colchicin revealed a close relation of the pinealocytes secretion with their electrical activity. Existence of central (olfactory in particular) mechanisms limiting the pineal activation, was shown. PMID- 11059022 TI - [Physiology of secretory cells in the mouse mammary gland]. AB - Secretory cells' membrane potential and transepithelial potential difference in the mouse mammary gland diminish within 2.5 hours following breast-feeding of the litter. The transepithelial resistance for up to 20 hours after the feeding did not drop below 40-70 k omega. The secret pressure in the mammary gland does not grow during this period. Therefore an increase of interval between litter feeding up to 20 hours does not entail any mechanical lesion of the secretory epithelium. The latter's cells seem to secrete organic and inorganic substances in concentrations which do not change significantly during their transfer along the outgoing ducts. PMID- 11059024 TI - [Cystic fibrosis]. PMID- 11059023 TI - [Response of membranes of mechanosensitive neurons in Astacus astacus to the intracellular injections of solutions under pressure]. AB - Injections of iso- and hypertonic solutions of some potassium salts caused an obvious depolarisation and reduction of electrical resistance in the crayfish slowly adapting stretch receptor neurones' membrane. This response could be eliminated with gadolinium. The stretch-activated channels in the crayfish stretch receptor neurone seem to be affected by an intracellular hydrostatic pressure. PMID- 11059025 TI - [Indications for lung transplantation in advanced cystic fibrosis]. AB - Lung transplantation has become a valid therapeutic option for cystic fibrosis patients with end-stage lung disease. The indication for transplantation does not rely on strict criteria only but must be evaluated case by case. In particular, the dynamics of the clinical course need to be considered with regard to impaired physical performance, recurrent infections, decline in pulmonary function and weight loss. Important risk factors are a poor nutritional status, osteoporosis, liver involvement, previous pleurodesis and the occurrence of multiresistant bacteria. Management and assessment of cystic fibrosis patients for lung transplantation is complex. Therefore patients should be referred to specialised centres at an early stage. PMID- 11059026 TI - [Early detection of exacerbation of lung infections in patients with cystic fibrosis by means of daily spirometry]. AB - The clinical course of cystic fibrosis is characterised by pulmonary involvement with mucus retention, chronic pulmonary infection and parenchymal inflammation. Recurrent infectious exacerbations are usually accompanied by a fall in lung volumes. This pilot study investigated whether exacerbations can be detected early by daily spirometry. Ten patients with cystic fibrosis (5 female; 5 male; mean age 24.9 years) performed daily spirometry using a portable transtelephonic spirometer (Spirophone). Infectious exacerbations were diagnosed on clinical grounds and treated without knowledge of the spirometry results. Data of 9 patients recorded over a period of 5-11 months were analysed. One patient was excluded due to non-compliance. A total of 20 infectious exacerbations occurred during the observation period. A fall of at least 20% in one or more of the following parameters was observed in 90% (18/20) of exacerbations: FVC, FEV1, PEF, and FEF25/75. A daily drop in lung volumes of 0.7% to 1.2% was recorded beginning at a median of 33 (20 to 120) days before infectious exacerbations were diagnosed. There was a 2-3% daily improvement in spirometric data under treatment with antibiotics. CONCLUSION: Daily spirometry allows early recognition of pulmonary infectious exacerbations in patients with cystic fibrosis. Daily spirometry may be used as an indicator for early antibiotic treatment. PMID- 11059027 TI - [Inhaled colistin in cystic fibrosis]. AB - The clinical course of cystic fibrosis (CF) is characterised by chronic bronchial infection with Pseudomonas aeruginosa. Therapy with inhaled aminoglycosides was introduced to decrease the rate of infectious exacerbations and to delay pulmonary progression. However, development of resistance to aminoglycosides is frequent. Few investigations are available into the resistance profile under treatment with colistin. Antibiotic resistance to colistin was analysed in 44 adult CF patients treated with inhaled colistin. Resistance to aminoglycosides was observed in 86% of cases (38/44) before therapy and decreased to 43% (19/44) under treatment with colistin. Five patients (11%) developed polymyxin resistance. After cessation of therapy pseudomonas became sensitive to polymyxin within a few months and enabled colistin to be reintroduced. In addition, we performed a pilot study analysing the effect of inhaled colistin on the growth of pseudomonas. The number of Pseudomonas aeruginosa decreased from 16.7 million (CFU) bacteria per ml sputum to 2.9 million under therapy with colistin. There was a more than tenfold increase in bacterial counts after inhaled colistin was stopped. Genotyping revealed no change in the type of pseudomonas strains. CONCLUSION: Development of resistance to polymyxin is not rare under long-term treatment with inhaled colistin and requires temporary interruption of therapy. Sputum cultures should therefore be tested regularly for polymyxin resistance in patients treated with inhaled colistin. PMID- 11059028 TI - [Swiss registry for patients with cystic fibrosis: design, programming, implementation and first examples of use]. AB - The Swiss Registry for Cystic Fibrosis (SRCF) was designed to collect demographic, clinical and therapeutic data from patients with cystic fibrosis (CF) in Switzerland. It was designed, programmed and implemented for standalone application in Swiss cystic fibrosis centres. It is part of the European Registry for Cystic Fibrosis (ERCF), which has been implemented in Europe to collect data on the use and safety of dornase alpha (Pulmozyme) in the treatment of cystic fibrosis. At the time of first evaluation 245 cystic fibrosis patients are registered, their mean age is 13 years, and 17% are over 18. In larger databases in Germany or North America we observe comparable demographic data, similar degrees of severity and similar therapeutic approaches to those in Swiss cystic fibrosis patients. The aim of the Swiss Registry is to cover the maximum possible number of cystic fibrosis patients from this country. PMID- 11059029 TI - [When cats scratch]. PMID- 11059030 TI - First sexual intercourse and contraception: a cross-sectional survey on the sexuality of 16-20-year-olds in Switzerland. AB - BACKGROUND: The onset of sexual activity represents an important development stage with positive aspects, such as love, discovery, intimate relationships, plans for the future, but also with fears of pregnancy, of sexually transmitted diseases and of AIDS. OBJECTIVE: To study perceptions, attitudes, and behaviour related to sexual life, AIDS and contraception; to explore the onset of first sexual relationships and the process of choice of a contraceptive method by the adolescents, in order to improve prevention programmes for young people. METHOD: Analysis of data from a Swiss national survey on adolescent sexuality using a computerised self-administered questionnaire, involving 2075 girls and 2208 boys between the ages of 16 and 20. The use of computers helps improve confidentiality, response rates and acceptability since survey questions are limited to the subjects' sexual experience only. RESULTS: The young people's responses emphasised the importance of emotion in sexual relationships, girls choosing intimacy and fidelity while boys attached more importance to physical pleasure. Three out of four respondents have had a sexual experience and one out of two have had penetrative sexual intercourse. The percentages of condom or oral contraception use are high: at first sexual intercourse, 86.5% used one or the other, while 7.4% did not declare any contraceptive method. The percentages are lower when age at first intercourse is below 15 years, when a girl had an older partner (age difference 7 years and more) and when the 1st relationship is a casual one. During their first stable relationship 41.1% of girls and 30.9% of boys say they have changed their contraceptive method from condom to contraceptive pill, 2.4% of girls and 2.9% of boys say they have given up any form of contraception. Among girls, condom use at first sexual intercourse with a new partner decreases in favour of oral contraception between first and last steady relationships (75.6 vs 58.0%, p < 0.05), the decrease being insignificant between the first and last casual relationships (73.5 and 62.2%, n.s.). Among boys the rates of condom use are equal at first intercourse with the first and last partner (steady relationship: 74.1 and 77.2%; causal relationship: 78.3 and 76.2% respectively). CONCLUSION: The use of condoms is high among Swiss adolescents, particularly at first sexual intercourse. By integrating the prevention of sexually transmitted diseases and of unwanted pregnancies, preventive programmes would address adolescents' needs more effectively. PMID- 11059031 TI - [Laparoscopic fundoplication for gastroesophageal reflux: experience with 49 surgical patients (1994-1999)]. AB - INTRODUCTION: Chronic gastrooesophageal reflux disease (GERD) is the most common benign pathology of the upper gastrointestinal tract in the western world. We report our experience of laparoscopic antireflux surgery. PATIENTS AND METHODS: 49 patients underwent laparoscopic antireflux surgery at our clinic between 1994 and 1999. 48 patients were followed up in a retrospective study. Mean follow-up was 30 months (2-66). 31 patients (64.6%) were male and 17 female (35.4%). Mean age was 48 years (26-74). The surgical method was tailored to the case: total Nissen fundoplication (87.5%) was indicated after ruling out oesophageal motility disorders by manometry. In 6 patients (12.5%) with coexisting dysphagia or pathological manometry, partial posterior fundoplication (Toupet) was performed. RESULTS: Mean operating time of 215 minutes (125-420) for the first 10 Nissens was significantly reduced to 119 minutes (70-190) for the last 10 procedures with increasing experience of the surgeon. No severe intraoperative complications occurred and mortality was 0%. Conversion rate was 4.2%. Mean hospital stay was 6.1 days (1-36). At follow-up 93.7% were free of reflux symptoms without medication, and only one patient (2.1%) suffered from regular reflux which had to be treated with PPI daily. 2 patients (4.2%) took PPI only occasionally. Persistent dysphagia occurred in 7 patients (14.8%). 5 (10.5%) underwent one or more endoscopic dilatations, after which 3 patients (6.3%) reported an improvement of dysphagia. No patient needed reoperation on due to mechanical complications. 1 patient (2.1%) developed a paraoesophageal hernia 4.5 years after a Nissen procedure. According to the Visick Score, 95.8% of all patients were satisfied with their outcome (Visick I/II). CONCLUSIONS: With careful investigation and indication, laparoscopic antireflux surgery is a safe and effective alternative method to long-term medication with PPI in the treatment of gastrooesophageal reflux disease. Morbidity is low. Persistent postoperative dysphagia can be reduced with either a short and floppy total fundoplication or a partial wrap. PMID- 11059032 TI - [Late manifestation of radiation injury to the plexus brachialis and plexus lumbosacralis]. AB - Radiotherapy of breast cancer, cervical cancer, testicular tumours and lymphoma is one of the most effective therapy options. Damage to the nervous system, in particular the brachial and lumbar plexus, is rare and typically leads to development of progressive sensory disturbances and motor weakness after years long latency. We present two cases exemplifying the diagnostic problems in differentiating between radiation-induced injury and recurrence of the primary tumour. A clinical course with sensorimotor symptoms and signs progressing over months, electomyographic recording of myokymic discharges, and absence of a space occupying mass suggest late-onset radiation-induced plexopathy. The literature on pathogenesis and incidence of radiation-induced plexopathy is reviewed. PMID- 11059033 TI - [Hand-foot syndrome (palmar-plantar erythrodysesthesia]. AB - The hand-foot syndrome (HFS) is an erythematous skin lesion of the palma and planta of the hand and feet is most often caused by cytostatic chemotherapy. The impact on the patients' quality of life depends on the extent of the disease. The pathogenesis of the hand-foot syndrome has not yet been sufficiently clarified and it can only be treated symptomatically. We performed an extensive literature search in Medline to evaluate the current state of knowledge concerning the hand foot syndrome and conclude with practical advice to physicians treating patients with hand-foot syndrome. PMID- 11059034 TI - [On closer examination]. PMID- 11059035 TI - Comparison between comparative tuberculin and gamma-interferon tests for the diagnosis of bovine tuberculosis in Ethiopia. AB - A study to determine and compare the sensitivities and specificities of the comparative cervical tuberculin (CCT) and gamma-interferon (IFN-gamma) tests for the diagnosis of bovine tuberculosis was conducted on 30 zebu oxen. The results of the tests were compared with the presence of acid-fast bacilli found by bacteriological culturing and histopathological examinations. The sensitivity and specificity of CCT test were found to be 90.9% and 100%, respectively. Those of the commercial IFN-gamma test were determined to be 95.5% and 87.7%, respectively. No significant differences were found between the sensitivities (Yates' corrected chi 2 = 0.32; p = 0.57) or the specificities (Yates' corrected chi 2 = 2.54; p = 0.11) of the two tests. Furthermore, a positive correlation (r = 0.76) was recorded between the increase in skin thickness following injection of bovine purified protein derivative (PPD) and the optical density in the gamma interferon assay with bovine PPD. On the other hand, the correlation (r = 0.47) between the change in skin thickness following injection of avian PPD and the optical density in the gamma-interferon assay with avian PPD was relatively weak. On the basis of this preliminary investigation, it was concluded that the choice between the two tests depends on their cost and simplicity and on livestock management and time factors rather than on their respective diagnostic value. PMID- 11059036 TI - Response of chickens to infection with Newcastle disease virus isolated from a guinea fowl. AB - An isolate of Newcastle disease virus obtained from a guinea fowl was characterized as a viscerotropic velogenic strain based upon pathogenicity index studies. Following inoculation of the viral isolate oronasally into 3-week-old chickens, clinical signs appeared after an incubation period of 4-5 days and included dullness, depression, dyspnoea, diarrhoea and leg paralysis. The virus caused a mortality of 56% with haemorrhages at the tip of the glands of the proventriculus and caecal tonsil. Histopathological changes were prominent in the lymphoid organs, being characterized by depletion, degeneration and necrosis of the lymphoid tissues. The brain was the first organ affected, with changes being noticed 3 days after infection. Isolation of virus from various organs was more frequent from 5 to 10 days after infection, but the virus persisted in some of the organs until 21 days after infection. In spite of the high mortality, a good immune response was elicited by the isolate, as was evident from the antibody titre. PMID- 11059037 TI - The role of management segregations in controlling intra-herd foot-and-mouth disease. AB - Transmission of foot-and-mouth disease virus (FMDV) by aerosol spread can occur over considerable distances. However, this is less effective in hot, dry environmental conditions, and a detailed study of an outbreak within a large dairy herd in Saudi Arabia has shown that contact spread is the main mode of transmission within a herd: both physical and spatial barriers curtailed the course of disease across the farm. Hence, the speed and path of an outbreak can be altered by changing the positioning of spatial or physical barriers. Extending the distances between pens, increasing the number of farm pens, decreasing the number of animals within the pens, and placing pens of well-protected stock between those of susceptible stock, can all contribute to the control of FMD involving contact and short-distance aerosol spread. Such management techniques offer a cost-effective supplement to control by vaccination. PMID- 11059038 TI - The effect of strategic anthelmintic treatment on internal parasites in communally grazed sheep in a semi-arid area as reflected in the faecal nematode egg count. AB - Communally grazed sheep were dosed at 4-, 12-, 24- or 48-week intervals for 1 year. Dosing every 4 weeks proved to be the most effective (p < 0.05), as reflected in a lower worm egg count compared to the 12-, 24- or 48-week intervals. Since most nematode life cycles lie between 3 and 6 weeks, the treatment has to be given during this critical period if maximum economic advantage is to be gained from deworming. However, treating communally grazed sheep every 12 weeks was found to keep worm egg numbers relatively low and may be advantageous in providing seasonal control, especially in semi-arid environments. Dosing communally grazed sheep once or twice a year under the same conditions is not recommended because reinfection appeared to result in similar faecal egg counts to those from the untreated animals (p > 0.05). PMID- 11059039 TI - Transmission of theileriosis in the traditional farming sector in the southern province of Zambia during 1995-1996. AB - The incidence of first contact with the protozoan Theileria parva was determined in three traditional cattle herds in the Southern Province of Zambia in 1995 and 1996. The majority of first contacts occurred during the dry season in June, July and August, at a time of nymphal activity and in the absence of Rhipicephalus appendiculatus adults, indicating that larva to nymph transmission plays a more prominent role than nymph to adult transmission under the prevailing conditions. PMID- 11059040 TI - Influence of minerals on the aetiology of geophagia in periurban dairy cattle in the derived savannah of Nigeria. AB - The aetiology of geophagia in periurban dairy cattle in Nigeria was assessed in relation to the mineral status of the serum, the herbage consumed, and the soil from where the herbage was consumed. The study was carried out using nine herds in the derived savannah zone during both the wet and dry seasons. Generally, the Na, Ca, Fe, Zn and Cu levels in the samples were adequate. However, the forages were low in Ca and the serum was deficient in P. Forage, soil and termite hill samples were also deficient in P. The low levels of P in the soil and termite hill samples showed that the consumption of sand by the cattle did not have any positive impact on their P intake. A seasonal effect was apparent (p < 0.05) on the serum Na+, PO4(3-), Cu2+ and Fe2+ concentrations but not on Ca2+ and Zn2+ concentrations. A seasonal effect was also significant (p < 0.05) on the minerals in the forage. There were also seasonal differences in most of the soil and termite hill minerals. Most samples had higher (p < 0.05) mineral levels in the dry season. Herd size and farming activity had no apparent influence on the aetiology of geophagia as they did not affect (p > 0.05) the concentration of P in the serum, the reported cause of geophagia. Geophagia is probably caused by a marginal level of P in the serum and low levels in the forage and soil. Experimental trials will be required to confirm these observations. PMID- 11059041 TI - Laying performance of broiler breeder chickens fed various millets or broken rice as a source of energy at a constant nutrient intake. AB - Laying performance, egg quality and carcase traits were recorded in broiler breeders (29-48 weeks of age) fed diets in which pearl millet (Pennisetum typhoides), finger millet (Eleusine coracana), foxtail millet (Setaria italica) or broken rice fully replaced yellow maize (50% of the diet) from the reference diet. Constant ratios of metabolizable energy to other nutrients were maintained in all the diets. Each diet provided 1.38 MJ metabolizable energy (ME)/bird per day to 8 replicate groups of 10 layers each, maintained in deep litter pens. The hen-day egg production and the ME required to produce 12 eggs in birds fed pearl millet or broken rice were similar (p > 0.05) to those fed on the maize diet, while birds fed foxtail millet or finger millet required more energy to produce the same number of eggs. The egg production and the efficiency of energy utilization were significantly (p < 0.05) reduced in the birds fed these two cereals. Egg quality was not affected by replacing maize with the different cereals. However, the yolk colour index was significantly (p < 0.05) reduced in the cereal-fed groups in comparison to the maize-fed birds. The gizzard and giblet weights were heavier (p < 0.05) in the birds fed foxtail millet or finger millet than in those fed broken rice. Deposition of abdominal fat and the liver fat content were significantly (p < 0.05) higher in the birds fed pearl millet than in the other groups. Depending on the local prices, the cost of feeding may be reduced considerably by using any of the cereals tested as the principal energy source in place of maize in broiler breeder diets. PMID- 11059042 TI - The value of vascularized tendon transfers with free flaps in perioral and cheek reconstructions. AB - The effect of gravity and the process of ageing necessitate the support of flaps used for the reconstruction of facial tissues. Perioral and cheek reconstructions exemplify this problem. Support can be obtained using vascularized tendons with free tissue transfers. The radial-forearm neurocutaneous flap with the vascularized palmaris longus tendon and the neurocutaneous lateral arm flap with the vascularized triceps tendon are useful options for resurfacing of these defects. We present 6 cases to highlight the use of the vascularized tendon transfer with free neurocutaneous tissue transfers as composite flaps in perioral and cheek reconstructions. PMID- 11059043 TI - Capsular contracture in augmentation mammaplasty. AB - In a group of 331 augmentation mammaplasties performed at the Clinic of Plastic Surgery in Prague from 1994-1998, capsular contracture was recorded in 6% cosmetic and 12% of reconstruction operations. The main cause was, in particular, the quality of tissue surrounding the implants along with individual disposition. The incidence of this complication did not differ significantly in relation to the placement of the prosthesis beneath the muscle or gland. Based on histological analysis and the clinical finding, it is obvious that disintegration of the prosthesis does not always lead to severe grades of capsular contracture. It is despite the presence of foreign bodies in the connective tissue. PMID- 11059044 TI - Alternative method for the reconstruction of defects with a loss of more than half of the upper eyelid. AB - The authors wish to present an alternative technique for the reconstruction of defects involving the loss of more than half of the upper lid. In a situation where the standard techniques could not be used due to the specific nature of the local findings, the described method seems to be a sound alternative for lid reconstruction. PMID- 11059045 TI - Hourly diuresis in patients with extensive burns. AB - The values of hourly diuresis in 40 patients with extensive burns were investigated and expressed graphically during the stage of burn shock. By analysis of the values and the patient's condition, a marked effect of the mental state on the course of burn shock and further treatment at the intensive care unit of the Prague Burns Centre was found. On a preliminary basis the effects of some drugs on the values of hourly diuresis were investigated. PMID- 11059046 TI - Marjolin's ulcer. AB - In 1828 John Nicolas Marjolini characterized ulcer with malignant degeneration which developed in scars after burns, but it occurs under varying clinical conditions. Typical feature is the latent period (on average 30 years). It is encountered in 2 forms: a shallow ulcer or exophytic tumour, most frequently on the lower extremities. At the Prague Burn Centre 11 patients were treated since 1978 till 1998. A unique case was a man suffering from congenital form of epidermolysis bullosa who developed Marjolin's ulcer on his foot (histological examination confirmed well differentiated squamous cell carcinoma). After 4 years he died with extreme cachexia and metastatic spread of the tumour, because he refused repeatedly amputation and lymph nodes dissection. To prevent Marjolin's ulcer several recommendations are presented. PMID- 11059047 TI - Poland-Mobius syndrome and disruption spectrum affecting the face and extremities: a review paper and presentation of five cases. AB - The author summarizes hitherto assembled experience with the clinical and genetic characteristics of Poland's and Mobius syndrome. Five selected case-records with this disease and the sequence of the Poland-Mobius syndrome are presented. Another case-record is devoted to an allied syndrome, hypoglossia-hypodactyly, found in a spontaneously aborted fetus. For establishment of a more accurate symptomatology, an irreplaceable place is held by anthropometric examination; for objectifying the asymmetry of the chest the so-called cyrtogram, the chest circumference recorded by means of a wire, is valuable. From the aspect of genetic counseling, preconception care is always provided to mothers from families with reproductive intentions, as well as ultrasonographic examination of the fetus in areas of assumed acral symptomatology (signaling phenotype). In two families ultrasonography was used for prenatal diagnosis. Invasive prenatal diagnosis by amniocentesis was employed in a family with Mobius syndrome. In these families dermatoglyphs have certain common characteristics, such a tendency towards simple patterns. In the wider family of one of our patients we detected in a cousin Parkes-Weber-Klippel-Trenaunay's syndrome, which may indicate common vascular predisposing factors. PMID- 11059048 TI - [Relationship between obesity and educational level in Portuguese young males in 1990]. AB - OBJECTIVE: To evaluate the relationship between the educational level, weight and obesity of young Portuguese males, aged 20 years. DESIGN: Cross-sectional study, based on the national military census files of 1990, provided by the military selection centres of Portugal (north, centre, south--including Azores and Madeira islands). SUBJECTS AND METHODS: In Portugal, all males aged 20 must present themselves to the military authorities to be submitted to a health exam in order to be screened for the armed forces. The data of 70,858 young Portuguese males aged 20 were gathered. Height and weight were measured by skilled teams. The educational level was confirmed by the presentation of educational attainment certificates. The population was divided into deciles of body mass index (BMI). The whole group was classified as obese or non-obese, according to a cut-off point of 27.8 Kg/sqm. The educational level was classified in four groups: group A--less than 5 years of schooling; group B--5 to 6 years; group C--7 to 9 years; group D--> or = 10 years. The level of education in each decile of BMI was determined and the percentage of obese subjects was calculated for each level of schooling. The odds of obesity associated with each group of schooling was determined as well as their 95% confidence intervals. Statistical significance was considered to exist when p < 0.05. RESULTS: The average BMI in the study population was 22.5 Kg/sqm. A slight increase in schooling was noticed from the lowest to the highest decile of BMI. The same result was obtained when we analysed the upper extreme of the BMI distribution in greater detail. According to our criteria for obesity, the percentages of obesity in each level of schooling were as follows: group A--4.2%; group B--5%; group C--5.1%; and group D -5.4%. The odds ratio (OR) of obesity according to each education level was higher in those with a higher level of education. In fact, those with 10 or more years of schooling presented an OR = 1.05(0.94;1.17) which was not statistically significant (ns) when compared to those with 7-9 years; while an OR = 1.08(0.97;1.21), was also ns when compared to those with 5-6 years, and, finally, an OR = 1.29(1.14;1.47) was statistically significant (p < 0.001) in comparison to those with only 0-4 years of schooling. CONCLUSIONS: As major conclusions one can say that the prevalence of obesity is low among young Portuguese males (4.9%), only 2.1% with a BMI > 30 Kg/sqm. Furthermore, our results show that, in Portugal, contrary to the results found mostly in western European countries, males aged 20 years with higher levels of education are slightly more prone to be obese than others with lower levels of schooling. PMID- 11059049 TI - [Efficacy and safety of diltiazem in the treatment of arterial hypertension in the elderly]. AB - A multicentric, non-comparative study was made to evaluate the efficacy and safety of diltiazem 180 mg in the elderly, with dose titration in subjects above the age of 60 years. The blood pressure measurements were done by ambulatory blood pressure monitoring (ABPM). We achieved a reduction of at least 10 mmHg in systolic blood pressure in 52.2% and in diastolic blood pressure in 42.5% of the patients. The estimated differences between the initial values and those after treatment are 30% (CI95%: 22-38%) for diurnal blood pressure load, 22% (CI95%: 14 29%) for nocturnal blood pressure load, 13 mmHg (CI95%: 10-16 mmHg) for median systolic BP, and 8 mmHg (CI95%: 6-10 mmHg) for median diastolic BP. In 62.5% of the subjects the daily dose of diltiazem was increased to 240 mg (180 mg in the morning and 60 mg at night). All the cardiovascular parameters evaluated showed a significant decrease at the end of the study. A significant change was not recorded in the laboratory parameters and the adverse event average was minimal. The results showed that diltiazem in monotherapy is effective in the control of hypertension in the elderly and can improve compliance to the treatment. PMID- 11059050 TI - [Leprosy. Comparative study of old and new patients]. AB - Leprosy represented, until a few decades ago, an important public health problem in mainland Portugal. According to the data of the General Directorate of Primary Health Care, the incidence and prevalence rates of the disease have been decreasing progressively and in a significant way during the last years. In view of the fact that it is an endemic disease in regression, we decided to estimate if there were significative changes in the clinical manifestations of the new cases. Therefore, we carried out a comparative study based on the clinical data of 18 patients (13 males and 5 females) with Leprosy who were being followed up in the Dermatology Clinics of Coimbra University Hospital. We differentiated two groups: patients with the diagnosis of the disease before 1990 (old patients), and those diagnosed after 1990 (new patients). The first group had only 8 subjects while the second group had the other 10. We recorded the patients' age, sex, nationality and residence, age at the onset of the disease, number of cases that began before the age of 20, clinical manifestations (according to Ridley and Jopling's classification), and the presence of incapacity determined by the consequences of the disease. We carried out the Student's-t test to appreciate some of the study parameters. We verified that the old patients were on average 11 years younger than the new patients. Leprosy in new patients occurred 11 years later than in the old patients and this difference was statistically significant after the Student's-t test (p < 0.05). Almost one half of the new cases (4 patients in 10) were possible cases of leprosy brought from Brazil, while all the old individuals had an autochthonous origin. The multibacillary forms were predominant in both groups, representing 89% of all patients. The frequency of handicaps was similar in both groups. Although it is based on a small number of patients, our study suggests that the clinical and epidemiological manifestations of leprosy have been changing significantly in the last years and that the changes observed in the form of presentation of the new cases are consistent with the present epidemiological situation. Our study also seems to suggest an increase in the importance of leprosy imported by immigrants from leprosy endemic countries. We point out that this phenomenon, as in the case of AIDS, can theoretically create the conditions for a recrudescence of leprosy, as currently observed with tuberculosis. PMID- 11059051 TI - [Menisci and posture]. AB - The first aim of this work is not only to review the localised perspective of meniscopathy, concerned with the consequences of meniscectomy, but to also view it in a broader dimension, in the behavioural aspect--related to postural activity. The second aim is to establish the relationship between these two dimensions. Meniscopathies invariably lead to degenerative alterations of the knee joint--not sufficiently explained by the local factors--that result in a situation of osteoarthritis. Some investigators established that the osteoarthritis process should not be confined only to the mechanical responsibility, due to some studies that also confirm the existence of biochemical alterations. However, others have also shown that the nervous system (NS) is likely to influence the inflammatory manifestations through the unmyelinated afferent fibers and sympathetic efferent fibers of the joints. These fibers can interact with non-neural elements, releasing some mediators, such as P substance (PS) and norepinephrine (NE), which, by themselves, or through other substances, contribute to the exacerbation of the inflammatory process. In order to relate the facts above, this longitudinal study comprised the following approaches clinical: anthropometric; biotechnical; and posturographic. It was characterised by five moments of data collection, the periodicity of which is related to the time of the surgery: the first moment is before surgery, followed by the remaining four, at six-week intervals, the sample being composed of--15 male caucasians, aged between 20 and 30 years, working for the Air Force. These Subjects were divided into two groups, according to the amount of meniscus removed in the longitudinal direction. Group A--meniscectomy < 1/2 the longitudinal body, composed of 7 subjects, with an average age of 21.4 years; and Group B, meniscectomy > 1/2 the longitudinal body, composed of: 8 subjects, with an average age of 24.1 years. The statistical analysis contained a descriptive approach that was comprised of--the average, the standard deviation and the limits of variation or variation range--and an inferential approach--ANOVA-One Way. The error probability chosen was p < or = 0.05. The tendency to recover was shown from the 3rd moment onwards (12th week), muscular involution also being implied by an involution of orthostatic postural activity, which confirms the interdependence between muscular tonicity and postural activity. In what concerns metabolic alterations, there was no parallel evolution with the other variables since, when the signs of recovery of the latter were verified, the osteo cartilaginous manifestations of destruction and reconstruction are more emphasised. On the other hand, the manifestations of postural activity reach their highest values before the manifestations protagonized by biochemical substances related to osteo-articular alterations. The accuracy of the posturographic tests for the diagnosis of deficiencies in what concerns of motor activity was shown, such as the inversion of the dislocation amplitudes on the YY' axis in relation to the XX', visible when the performance of the injured limb is evident, which leads us to admit that this fact is a consequence of meniscal pathology. PMID- 11059052 TI - [Mandatory notification of communicable diseases: what physicians think]. AB - OBJECTIVE: To find out the opinion of family doctors concerning the statutory notification of communicable diseases (SNCD) and to analyse factors associated to a good attitude towards SNCD. SETTING: Guimaraes Health Centre, Guimaraes. METHODS: Analytical cross-sectional study. Questionnaire survey. The 52 family doctors of the Health Centre were asked to express their opinion on some issues, including the utility of SNCD, reasons for the under-reporting of communicable diseases, up-to-dateness of the list of notifiable conditions, facility of filling in the notification form, and their own attitude towards notification. The variables utility of SNCD and attitude towards notification were then considered together with other variables (including postgraduate time, length of the family doctor's lists of patients, working in exclusivity, and how the degree of general practitioner is obtained). RESULTS: The SNCD was considered at least somewhat useful by 96% of the doctors. Thirty percent of the doctors considered excess work and/or lack of time as the main reason for under-reporting, and 29% it attributed to lack of sufficient motivation. The list of notifiable conditions was considered out dated by 46% of the doctors. Thirty eight percent of them considered the multiple notification of a case of disease not inconvenient and 54% stated that they had used it to notify all or almost all of cases they knew. The doctors who worked exclusively as civil servants and who had smaller lists of patients seemed to have a greater compliance regarding notification (p < 0.05). CONCLUSIONS: The majority of doctors considered the notification useful. They attributed the under-reporting to some conditions that are difficult to change. In this population, the doctors who worked exclusively as civil servants and who had smaller lists of patients showed a more favourable attitude towards reporting notifiable conditions. PMID- 11059053 TI - [Iatrogenic vascular injuries]. AB - Iatrogenic vascular injuries include an extremely broad group of lesions with multiple etiology and polymorphic manifestations. We present a series of 44 patients with iatrogenic vascular lesions that result from arterial and venous catheterizations and surgical interventions. The arterial catheterization for coronariography and arteriography and orthopaedic surgery were the agents that most frequently caused vascular injury. Arterial and venous reconstructive surgery were the predominant treatment procedures. In our series, morbidity and mortality were 4.6% and 2.3%, respectively. PMID- 11059054 TI - [Kikuchi and Fujimoto disease]. AB - We present a case of Kikuchi and Fujimoto's disease (KFD), revealed by malar rash, fever, arthritis and lymphadenopathy, suggesting Systemic Lupus Erythematosus (SLE). The relation between KFD and SLE is not yet completely understood and remains complex. There are few cases with a lupus-like presentation and the histopathologic features may be insufficient to confirm the KFD diagnosis definitively. The clinical presentation, the IgM deposits at the dermal-epidermal transition in the Lupus Band Test, and the presence of anticardiolipin antibody emphasises the relationship between these two entities and does not exclude the possibility of evolution to SLE, which makes the prognosis doubtful. PMID- 11059055 TI - [Rhodococcus equi pneumonia in patients with AIDS]. AB - Rhodococus equi infection is frequent in animals, but rare in humans. It usually appears as an opportunistic infection, occurring in clinical settings where a compromise of cellular immunity exists, such as in leukemia, neoplastic or transplanted patients. It is currently considered, as another opportunistic AIDS infection. Sixty six cases of Rhodococus equi pneumonia have been published until March 1996. The differential diagnosis includes TB and Staphyloccocal Pneumonia and is generally an infection which is difficult to treat, often resistant to Betalactamic drugs. Therapeutic options include an association of intracellular active antibiotics. The authors report the first Rhodococus equi pneumonia diagnosed in their Hospital and discuss the main difficulties in the diagnostic and therapeutic approach. PMID- 11059056 TI - [Development of the Portuguese version of MOS SF-36. Part I. Cultural and linguistic adaptation]. AB - No one aims at applying generic measures as substitutes for other more traditional clinical procedures. The whole history of the evolution of these types of measures has been based on comparisons with clinical measures, always seen by researchers as ways to validate health outcome measures and as a process to be recognized by clinicians as a way to detect changes in time not always detected by the usual measures. The measurement instrument presented in this paper is the Portuguese version of the MOS SF-36, originally a result of the Medical Outcomes Study, a study carried out by Rand Corporation researchers in the 80's. One of the objectives of these researchers was precisely to develop instruments to be used in continuous monitoring of outcomes. This paper describes the first time MOS SF-36 was culturally adapted to Portuguese, validated and implemented. The first part mentions some of the foundations and developments of the original instrument as well as some results obtained from some specific applications. The second part introduces operational definitions for each of the eight scales and describes the SF-36 measurement model as well as the factor structure with two dimensions. Next, we present the design used by us to transform the data from the time they are collected from the respondents to the time they are ready to be further used. Finally, the methodology used to culturally adapt the MOS SF-36 and create a Portuguese version which is culturally equivalent are presented. PMID- 11059057 TI - [Intermediate thalassemia: a new condition associated with hypercoagulability]. PMID- 11059058 TI - [Analysis of sperm aneuploidy in infertile subjects after chemotherapy treatment]. AB - The continuing search for a cure for cancer has developed more aggressive therapies that may damage germ cells, leading to clinical disease in offspring of survivors. Standard therapy for the majority of cancer today consists in combinations of high doses of radiation and chemotherapy drugs. We investigated the effect of cancer treatments on the reproductive potential of men. Multicolor fluorescence in situ hybridization has been used to recognize chromosomes X, Y and 8 in sperm of 10 severely oligozoospermic subjects (sperm concentration < 5,000,000/mL) treated for cancer at least 5 years before the beginning of this study. As controls, we analyzed sperm aneuploidies in 20 fertile men (sperm concentration > 20,000,000/mL) and in 20 severe idiopathic oligozoospermic subjects (sperm concentration < 5,000,000/mL). In all subjects, X- and Y-bearing spermatozoa were present in a normal 1:1 ratio; nevertheless the frequency of 24,XY, 24,XX and 24,YY disomic sperm was significantly higher in patients treated for cancer and in idiopathic oligozoospermic subjects with respect to normozoospermic men. These results suggest that the increase in sperm aneuploidies in treated patients cannot be reported directly to precedent chemotherapy, but reflects the alteration of testicular structure, as in the case of severe idiopathic oligozoospermic subjects. With the advent of intra cytoplasmic sperm injection, it is possible to offer the opportunity to conceive in men affected by severe oligozoospermia but it is also possible, when the spermatozoa of these subjects are used, to pass sex chromosome abnormalities on to the children. We therefore suggest caution before application of an artificial reproductive technique in severe oligozoospermic patients. PMID- 11059059 TI - Activated protein C resistance is a risk factor for central retinal vein occlusion. AB - Central retinal vein occlusion is one of the most common retinal vascular disorders. Few and contrasting data are available on the prevalence of hemostatic risk factors in patients with central retinal vein occlusion. The aim of this study was to investigate the most common hemostasis-related inherited risk factors for venous thrombosis in a group of 53 central retinal vein occlusion patients (median age 59 years, range 18-77 years) and in 53 comparable control subjects (median age 57 years, range 22-84 years). No difference was found in antithrombin III, protein C and protein S plasma levels between patients and controls. At univariate analysis, activated protein C resistance (odds ratio 5.8) and factor V Leiden (odds ratio 4.4) were significantly associated with central retinal vein occlusion whereas G20210A polymorphism of the prothrombin gene was not. After adjustment for sex, age, and the other classic vascular risk factors (hypertension, diabetes, hypercholesterolemia, smoking) activated protein C resistance remained the only independent risk factor for central retinal vein occlusion (odds ratio 11.5). These data indicate that activated protein C resistance may play a role in the pathophysiology of central retinal vein occlusion. PMID- 11059060 TI - [Pathogenetic and therapeutic aspects of secondary anorexia]. AB - Anorexia is an often underrated symptom in the clinical management of patients suffering from chronic diseases. Moreover, the anorexia accompanying chronic diseases (secondary anorexia) is often confused with anorexia nervosa, a typically neuropsychiatric disorder involving completely different pathogenic mechanisms and therapeutic strategies. Secondary anorexia is one of the main factors responsible for the development of malnutrition, which in turn negatively affects patient morbidity and mortality. Different mechanisms have been proposed to explain the pathogenesis of secondary anorexia. However, consistent experimental and clinical evidence seems to point to hypothalamic serotonergic system hyperactivity as a preeminent cause; this hyperactivity appears to be triggered by enhanced brain availability of tryptophan, the aminoacid precursor of serotonin. The hyperactive hypothalamic serotonergic system might also represent the final effector where different regulatory and modulating pathways, including cytokines, converge. The involvement of tryptophan and the hypothalamic serotonergic system is further supported by the effectiveness of a therapeutic strategy, based on the inhibition of tryptophan entry into the brain, in increasing the food intake of anorectic patients. Although these results represent an encouraging approach to the treatment of secondary anorexia, with possible beneficial effects on the nutritional status of patients, they need to be validated in larger trials. PMID- 11059061 TI - [Molecular medicine: new tools for better understanding and treatment of diseases in humans]. AB - The authors discuss the importance that molecular medicine has assumed in recent years. Molecular methodologies have clearly demonstrated that immunological diversity is based fundamentally on the rearrangement of the genes encoding antigen B and T cell receptors. The importance of oncogenes, and their translocation in tumoral pathologies is emphasized, a case in point being the alterations observed in chronic myeloid leukemia and acute promyelocytic leukemia and their implication for innovative therapy. The importance of prothrombin and factor V genetic-molecular alterations in thromboembolic pathology and of the activation of calcineurin phosphatase or other intracellular signal regulator molecules during cardiac insufficiency genesis is also discussed. Particular attention is paid to progress regarding the socially important Alzheimer's syndrome, and the diagnosis of endocrine tumors. Moreover, the authors believe that the identification of new endocrine nuclear receptors, "orphans" of hormonal ligands, will open up interesting prospects--even therapeutic--in endocrinology. The authors conclude by reviewing the therapeutic prospects for immunodeficiency syndromes and malignant tumors, offered by new gene therapy methodologies. They also discuss recent results of studies on the aging process which, until not many years ago, appeared adventuristic. Today they are opening prospects of great interest. PMID- 11059062 TI - [Possibly ranitidine-induced autoimmune hepatitis]. AB - Approved for clinical use in peptic ulcer in 1983, ranitidine competitively inhibits the interaction of histamine with H2 receptors. The incidence of adverse reactions has been low and generally minor. After reviewing the literature on hepatitis associated with ranitidine use, we report the case of a young woman, affected by multiple sclerosis, who developed severe liver injury associated with the intake of this drug. This case meets the principal criteria necessary to establish a causal relationship between the administration of a drug and the verification of an adverse reaction. The singularity of our case with respect to others reported in the literature arises from the demonstration of an anatomo pathological picture suggestive of toxic or idiosyncratic hepatitis after the initial administration that, at rechallenge, evolved into a histologic picture of autoimmune hepatitis. We believe that physicians should be aware of the potential hepatotoxicity of ranitidine. PMID- 11059063 TI - A case of pulmonary thromboembolism in thalassemia intermedia: are these patients at risk for thrombotic events? AB - We describe a case of pulmonary thromboembolism in a 48-year-old woman with thalassemia intermedia and no other risk factors. Multiple bilateral defects were detected by perfusion lung scan. No sources of emboli were detected, despite extensive evaluation. We suggest that a chronic hypercoagulable state due to multiple coagulation alterations might be a cause of thromboembolic events in thalassemia intermedia patients, even when no other risk factors are present. PMID- 11059064 TI - [Significance of "minor" genetic mutations in hereditary hemochromatosis: 2 case reports]. AB - We describe a case of homozygosity due to the substitution of aspartic acid with histidine at position 63 of the protein encoded by the gene (known as HFE) associated with hereditary hemochromatosis. Liver biopsy did not disclose stainable iron accumulation; serum ferritin was elevated (639 ng/mL), while the transferrin saturation index was within the normal range (38.1%). As the patient was affected by chronic hepatitis C virus, the high serum ferritin could be attributed to this disease, a frequent occurrence. We also describe a case of heterozygosity for both the substitution of tyrosine with cysteine at position 282 and the substitution of histidine to aspartic acid at position 63 (so-called "compound heterozygosity"). The patient had the typical biochemical abnormalities of iron overload: transferrin saturation index of 53.1% and elevated serum ferritin (658 ng/mL). The removal of > 5 g of iron by phlebotomies did not precipitate iron deficiency. Although the patient refused to undergo liver biopsy, clinical evidence alone enabled a diagnosis of hemochromatosis. These two cases concord with the present scientific orientation, i.e.: 1) homozygosity for the major mutation is associated with the phenotypical (clinical) picture of hemochromatosis, but compound heterozygosity also determines significant iron metabolism abnormalities; 2) homozygosity for the minor mutation does not appear to determine important phenotypical abnormalities. PMID- 11059065 TI - [Costal osteolytic lesions and multiple hepatic neoformations in a patient with inflammatory pseudotumor]. AB - Inflammatory pseudotumor is a rare disease, that is regarded as a benign reactive inflammatory process, although its etiology and pathogenesis are still unknown. The liver is one of the organs most frequently involved, but inflammatory pseudotumors have been reported in many other sites in the body. Inflammatory pseudotumor of the liver presents as a solitary or, less frequently, multiple space-occupying lesion, which the common imaging techniques do not clearly distinguish from primitive or metastatic hepatic malignancies. Biopsy of the lesion is therefore necessary for diagnosis. The case of inflammatory pseudotumor described here presented with radiologic features of multiple solid space occupying lesions in the liver, associated with multiple osteolytic lesions in the ribs. Such an association, very suggestive of malignancy, has not yet been reported for inflammatory pseudotumors. Optimum management of this disease has not yet been standardized. The majority of patients are treated by hepatic resection, although spontaneous regression has also been described. In our case, rapid improvement of both hepatic and costal lesions was observed, although the patient did not receive any specific treatment. PMID- 11059066 TI - [In Process Citation] PMID- 11059067 TI - Grain surface-layer treatment of diatomaceous earth for insect control. AB - This paper describes an alternative method to synthetic insecticides used for protection of stored agricultural products the purpose of which is to minimise the everyday human exposure to those chemicals. The method uses diatomaceous earth which is practically non-toxic to humans and fully acceptable for the environment. Fifty and 100-cm-deep layers of Hard Red Spring wheat Triticum aestivum (L.) in metal containers (cylinders), 30 cm in diameter and 150 cm in height were treated with 0.5 and 0.75 g of diatomaceous earth Protect-It per kg of wheat. The treatment reduced the population of Sitophilus oryzae (L.), Tribolium castaneum (Herbst) and Rhyzopertha dominica (Fabricius) by 98 to 100% with respect to controls. The conclusion is that a 100-cm-surface layer treated with 0.5 g/kg of Protect-It is sufficient to control these insects, and that no more than 20% of the total grain mass should be treated to minimise bulk density reduction. A field test using a similar design is essential to confirm the laboratory findings. PMID- 11059068 TI - Comparison of protocols for measuring activities of human blood cholinesterases by the Ellman method. AB - This paper presents a protocol for routine assays of human blood cholinesterase activities which separates erythrocytes from plasma by centrifugation and measures acetylcholinesterase activity in unwashed erythrocytes and butyrylcholinesterase activity in the plasma. The recommended substrate for both enzymes is 1.0 mM acetylthiocholine. The protocol is compared with other two recommended protocols for the activity measurements of the two enzymes using the Ellman method. The paper discusses the advantages and disadvantages of each and concludes with a proposal for an international agreement between laboratories for the evaluation of a standardized protocol. PMID- 11059069 TI - Screening of health care workers for hepatitis B virus and hepatitis C virus: criteria for fitness for work. AB - The aim of this study was to propose a protocol for assessment of markers of infection with hepatitis B virus (HBV) and hepatitis C virus (HCV) in exposed health care professionals and to define criteria for evaluation of fitness for the job of the infected personnel. The study comprised 800 persons involved in operative procedures, including 414 surgeons, 275 nurses, and 111 anaesthetists. A graduated protocol was created for monitoring markers of HBV and HCV infection. A well-defined combination of markers of antigen-antibody systems enabled identification of four groups of persons with HBV infection differing in fitness for work: 1) HBsAg-positive, HBeAg-positive, HBV DNA-positive; 2) HBsAg-positive, anti-HBe-positive, HBV DNA-positive; 3) HBsAg-positive, anti-HBe-positive, HBV DNA-negative; and 4) anti-HBs-positive, anti-HBc-positive, anti-HBe-positive group. For HCV infection, two groups with different job fitness were identified: 1) anti-HCV-positive, HCV RNA-negative and 2) anti-HCV-positive, HCV RNA positive. Screening of hospital personnel at risk to HBV and HCV infection requires a well-defined protocol which may help to evaluate the fitness of the infected personnel for a specific job. PMID- 11059070 TI - [Cancer of the respiratory organs with respect to environmental and life style factors in the areas of Split, Solina and Kastela] Citation]. AB - This investigation focused on Split and the industrial area of the Solin-Kastela bay, as it has been known for cement production for over 130 years, and for asbestos cement production for some 80 years. Environmental factors (air pollution) and lifestyle (cigarette smoking and alcohol consumption) were investigated in persons who died of cancer of respiratory organs. The aim was to see what is the combined effect of several noxious factors and how it may increase the risk of cancer of the respiratory organs. The sample of 473 subjects was randomly selected from 1490 persons who died of cancer of respiratory organs in the studied area between 1970 and 1990. The data on the subjects' lifespan, smoking habit, and alcohol consumption were obtained by interviewing their families. Environmental data showed that 5% had lived in an area with industrial air pollution (cement, asbestos-cement) for up to 15 years, 19% from 16 to 29 years, 33% from 30 to 49 years, and 44% for 50 years and over. The number of those exposed to air pollution for 50 years and over ranged from 36% to 68% in different districts. Of 473 subjects 85% had been cigarette smokers; 42% had smoked over 40 cigarettes a day, 83% had smoked for over 30 years, and 74% of subjects had consumed alcohol. The largest number of persons in the studied group were in the category with the longest exposure to air pollution and the longest and most intensive smoking experience. This indicates the possibility that simultaneous action of environmental factors and lifestyle may increase the risk for the development of cancer of respiratory organs. Smokers showed a significant shift (in percentage) towards shorter length of living in the studied area. That suggests that the risk of cancer of respiratory organs was greater in cigarette smokers than nonsmokers, regardless of the fact that both were exposed to comparable air pollution. PMID- 11059071 TI - [Biological effects of nonionizing radiation: low frequency electromagnetic fields]. AB - This article reviews various studies on effects of electric and magnetic fields of extremely low frequencies on human health and gives an overview of residential and occupational exposure to different sources, currently established exposure limitations, and protection measures. Throughout the evolution biological systems adapted to natural electric and magnetic fields. Only hundred years ago human exposure to radiation was limited to electric and magnetic fields arising either from extraterrestrial or terrestrial sources, yet both natural. For the past fifty years there has been large growth of artificial sources of electric and magnetic fields, especially with frequencies of 50 and 60 Hz (power generating and distribution systems). The concern about long-term exposure to artificial fields and possible adverse effects on human health has been entirely justified and led to numerous intensive epidemiological and laboratory studies. Results of several epidemiological studies confirm the connection between exposure to electric and magnetic fields of extremely low frequencies (up to 300 Hz) and increased risk of leukemia and brain tumor in children and adults. In addition, the risk of breast cancer in occupationally exposed population has increased. Laboratory studies on animal models, in vitro systems, and human volunteers did not confirm this connection. There is a growing interest in investigation of other possible adverse health effects such as neurodegenerative diseases (Alzheimer's disease and other forms of dementia, amyotrophic lateral sclerosis), cardiovascular disorders (arrhythmias and acute myocardial infarction), psychiatric disorders, and electrosensitivity. PMID- 11059072 TI - [Development of occupational medicine in Varazdin: from concept to creation]. AB - Since the antiquity and throughout the Middle Ages, physicians had been observing the harmful effects and the illnesses in people who worked in manufacture and/or mining. They would draw attention to such effects and propose solutions which often entailed improvement of working conditions and environment. The ancient physicians from Varazdin had been familiar with such literature, accepted the ideas thereof, and implemented them in practice. Moreover, they followed-up those who had been exposed to similar harmful effects. Some of their interpretations and understanding do not fall short of the modern perceptions of work-related harmful effects and some seem to be far ahead of their times. The introduction of administrative regulations was gradual; at start rare and related to exposure of certain individuals and later more frequent and comprehensive. Scientific approach to the need for systematic occupational safety has taken root in Croatia ever since World War I. Experts in the field have published important papers on the matter. Varazdin has joined the incentive for health care and protection of the exposed to harmful effects of modern industry and working environment since 1956. Physicians are specialising in occupational medicine. The section of occupational medicine in Varazdin was established within the Institute for Hygiene that later gradually transformed to a well organised unit for occupational health care. PMID- 11059073 TI - Sustainable rural practice. ...without a wife. PMID- 11059074 TI - Are evaluation and research mutually exclusive for divisions of general practice? PMID- 11059075 TI - Paying for consumer input. PMID- 11059076 TI - Immunisation and socioeconomic disadvantage. PMID- 11059077 TI - Antibiotic prescribing for URTIs. PMID- 11059078 TI - Changing hospital management of croup. What does this mean for general practice? AB - BACKGROUND: The hospital management of croup has altered significantly over the last decade, with current data suggesting that all children with croup who demonstrate an increase in difficulty breathing should be treated with corticosteroids, and children with more severe croup should be treated with nebulised adrenaline. OBJECTIVE: To discuss the assessment of croup severity, the recent changes in treatment and to make suggestions for the management of croup in general practice. DISCUSSION: Children with mild croup require reassurance. There is no evidence that steroids have a place in management in this group. A single dose of prednisolone is appropriate for children with stridor at rest, but no recession and they can be managed in the general practice setting provided they can be reviewed within 2-4 hours. Failure to improve after treatment with steroids means hospital referral. Children with more severe croup require hospital assessment and possible admission. PMID- 11059079 TI - Common corticosteroid injections. An anatomical and evidence based review. AB - BACKGROUND: The injection of depot corticosteroid preparations into soft tissues and joints has been used for some time to alleviate pain in a variety of musculoskeletal conditions. However the evidence, supporting the efficacy for these procedures, until recently, has been poor. OBJECTIVES: To review the recent literature on the efficacy and toxicity of commonly used corticosteroid injections in musculoskeletal medicine and to illustrate the key anatomy of the injection sites. DISCUSSION: Injections of corticosteroid into the lateral epicondyle, subacromial bursa, carpal tunnel, knee and plantar fascia all result in short term (weeks to months) alleviation of pain and other symptoms in the studies reviewed. The natural history of these conditions, however, is that the majority of patients improve over longer periods (months to a year) whether or not an injection has been given. PMID- 11059080 TI - A matter of minors. Minor procedures in children. AB - BACKGROUND: Laceration repair and removal of foreign bodies from the nose or ear in children are common procedures in family practice. Smaller anatomy and larger emotional response to these procedures in children can make the task challenging for even the most experienced practitioners. OBJECTIVE: This article revises some 'old tricks' and discusses some 'newer technologies' to aid the practitioner in dealing with these procedures. DISCUSSION: Proper preparation of the child is vital for a successful and distress free procedure. Enlisting the aid of parents is usually, but not always, helpful. Physical and/or pharmacological restraint should be used judiciously and only in a safe environment. A variety of methods of wound repair and foreign object removal are available for use in a general practice setting. The choice of method should be matched to the individual circumstances. PMID- 11059081 TI - Epistaxis. Strategies for management. AB - BACKGROUND: Epistaxis is a problem frequently encountered in general practice and may present as an emergency, as a chronic problem of recurrent bleeds or may be a symptom of a generalised disorder. OBJECTIVE: To provide practical guidelines for the management of the patient with epistaxis, with emphasis on the important aspects of patient history, examination and whether investigations are necessary. DISCUSSION: Given adequate knowledge of anatomy and clinical technique, epistaxis is easily managed. A focus on important aspects of history and examination assists in determining the site of bleeding and whether further investigations are necessary. Management includes resuscitation, if necessary, followed by cautery and/or packing of the site. PMID- 11059082 TI - Hypnotics. Options to help your patients stop. AB - BACKGROUND: There are a wide range of reasons why hypnotic agents may be started, but there is substantive evidence that they are often overused or misused. OBJECTIVE: To review the reasons for stopping hypnotic agents, how to detect their use, how to facilitate the decision to withdraw and how to most effectively achieve withdrawal. In this article, the target group is using but not intentionally abusing hypnotic agents. DISCUSSION: In many cases it is possible for patients to permanently stop using hypnotic drugs. Withdrawal can generally be achieved in the community, using a gradual taper regimen, although it is often a difficult process and is more effective when the patient is aware of the health benefits to be gained from withdrawal. Once withdrawn, other approaches to maintain sleep need to be reinforced to reduce the risk of recommencing hypnotic therapy. PMID- 11059083 TI - A case of mistaken identity. A rare presentation of gonadal dysgenesis. AB - BACKGROUND: The incidence of gonadal dysgenesis (hermaphroditism) is recognised to be low. Rarer still is an initial late presentation in the general practice setting. OBJECTIVE: To present a case study of a 35 year old man diagnosed as a hermaphrodite after routine investigations in general practice for lower abdominal pain. He has normal male external genitalia, a fully formed uterus and vagina, with no identifiable gonads. DISCUSSION: This incidental finding in general practice is supported by a 46,X,i(Yp)/45,X karyotype and mosaicism for an isochromosome of the short arm of the Y. It is not unusual that with normal male genitalia, such patients are likely to survive undiagnosed or incorrectly diagnosed into adulthood. PMID- 11059084 TI - Casting acute fractures. Part 2--Generic preparation and application of plaster slabs. AB - BACKGROUND: Slabs are ideal initial splints for the emergency treatment of many fractures. Construction of a broader slab of appropriate thickness with appropriate supportive elastic bandages produces a stronger, safer and more comfortable cast. OBJECTIVE: This article will introduce and demonstrate new materials and concepts that may be applied to slab construction in a generic fashion. It will form the basis for applying specific slabs demonstrated later in the series. DISCUSSION: The primary care physician is expected to apply slabs. Often his training has been minimal. The techniques shown are those used by full time casting technicians. These trade secrets are easily reproducible. Mastery will reduce material and labour costs for the general practitioner. The patient will have a stronger, more comfortable cast and fewer unplanned return visits. PMID- 11059085 TI - Geriatric medicine in practice. An overview. AB - Older patients with chronic health problems are a common presentation in general practice. The Central Coast Division of General Practice identified 'care of the elderly' as one of the priorities for local membership. This is the first in a series of articles based on the 'Geriatric Attachment Programme', which was jointly developed between the Division of General Practice and the Department of Geriatric Medicine, Central Coast Health. The recent introduction of the Health Assessment and Health Care Plans for the elderly into the Medical Benefit Schedule encourages preventive and anticipatory care. Doing this successfully requires a structured approach. This series will offer a practical guidance approach to enable readers to achieve this goal. In this article we provide an overview of the issues important to the care of the elderly. Subsequent articles deal with specific issues in detail. PMID- 11059086 TI - Growing older and wiser. Part 1--What is life all about? AB - This is an edited version of a presentation given at the 1999 RACGP Annual Scientific Meeting, held in Adelaide. Lady Valerie Hicks is the widow of Sir Stanton Hicks a renowned South Australian physiologist. As 1999 was the Year of the Older Person, Lady Valerie was asked for her reflections on what getting older means, the place of the older person in today's society and for her thoughts on life in general. Her erudite and thought provoking response to this task will be published over the next three issues. PMID- 11059087 TI - Choosing the most appropriate antibiotic for respiratory infections. PMID- 11059088 TI - Investigating a foreign body. Part 1--Where and how to look. PMID- 11059089 TI - Is this chronic asthma? PMID- 11059090 TI - Imaging changes in the ageing brain. PMID- 11059091 TI - To be or not to be. The practical aspects of contemplative practices--Part 2. PMID- 11059092 TI - Keeping up to date in the country. Finding the right balance. PMID- 11059093 TI - The learning preferences of rural and remote general practitioners. A quantitative analysis and its implications for the RACGP QA&CE program. AB - AIM: To establish the QA&CE needs of rural and remote GPs. In order to assess the fit between The Royal Australian College of General Practitioners' QA&CE program and these needs, learning preferences identified in this survey are compared to previous research published in Australia, 1987-1997, and matched against program objectives and guidelines. METHOD: In 1998 the RACGP surveyed 900 rural general practitioners (GPs) across Australia in three randomised samples. These samples were stratified respectively by membership of the Rural Faculty of the RACGP and practice location in RRMAs 3-7. The overall response rate was 78%. RESULTS: Self directed learning was rated by 65.3% of respondents as a preferred method of maintaining professional standards, a rating that corresponded significantly to familiarity with this approach to learning (p = 0.006), and even more so among younger GPs. This was second only to CME at 85.6%. By comparison, there was a low orientation toward some learning methods identified elsewhere as effective, such as clinical audit (15.6%) and peer review (14.5%). There were positive correlations, however, between preference and familiarity for these methods (p < 0.001). CONCLUSIONS: The results indicate that self directed learning, a major learning preference of rural and remote GPs, may currently be underserviced by the RACGP's QA&CE program. The program also needs to promote more strongly the effectiveness of less preferred modes of education, such as peer review and clinical audit, given the stronger preference indicated for less effective modes. PMID- 11059094 TI - Poisoning due to eating fungi in Victoria. AB - OBJECTIVES: To determine the range of fungi eaten in Victoria and the incidence and severity of associated illness. METHOD: From May 1997 to April 1999, 174 callers to the Victorian Poisons Information Centre who had eaten fungi posted samples for identification and 170 completed a questionnaire. The study was suspended for three months from 1 November 1997 to 10 January 1998 and for four months from 1 July 1998 to 30 October 1998. RESULTS: Species categorised as: 'poisonous', 'possibly poisonous', 'hallucinogenic', 'possibly hallucinogenic' and 'coprine containing' or 'possibly coprine containing' were identified in 87/174 (50%) samples. Accidental ingestions: 55 types of fungus were identified in the 126 ingestions; Coprinus species were the most common (24/126 [19%]). Illness 'likely' or 'possibly' due to the fungi was reported in 13/126 (10%) of these ingestions. Deliberate ingestions: The fungi were eaten for food in 46/47 of these cases; 41 of these (89%) were Agaricus xanthodermus or other Agaricus species. Illness 'likely' or 'possibly' due to the fungi was reported in 40/47 (85%) deliberate ingestions. In one case the reason for ingestion was unknown. CONCLUSIONS: A large range of fungi was eaten in the accidental ingestions; the incidence of illness was low. A small range of fungi was eaten deliberately. The predominant species was Agaricus xanthodermus, which was probably confused with other edible Agaricus species. The incidence of associated gastrointestinal irritation was high; it was of limited duration and mild severity. PMID- 11059095 TI - Handling information ethically. Some strategies for discussion. AB - BACKGROUND: A lack of consensus currently exists about which uses of patient data require informed consent or review by a human research ethics committee. However, any use of patient data other than for clinical care, requires the consent of the patient. OBJECTIVE: The aim of this paper is to discuss strategies for strengthening current practices regarding the use of patient information in general practice research, evaluation and audit. DISCUSSION: Increasing community discussion and debate with respect to the use of patient data for research and evaluation is a pressing issue. Divisions of general practice are well placed to lead in this debate given the extent of community representation in many divisions' activities. Development of guidelines for use by those undertaking research and evaluation activities in general practice, based upon the Privacy Principles, will ensure a high standard of protection for patients. Community based Human Research Ethics Committees may be an appropriate way of providing education and training as well as a review of general practice research and related activities. PMID- 11059096 TI - Low dose treatment with methotrexate-adverse drug reactions survey. AB - Antineoplastic drugs caused various and frequent adverse drug reactions (ADR) in connection with their pharmacodynamics. Methotrexate (MTX) ADRs are preferably gastrointestinal disorders and hepatotoxicity (hepatic enzyme abnormalities). The aim of this study was to detect and analyse ADR induced by low-dose MTX treatment in rheumatology. We observed 94 patients, 63 with rheumatoid arthritis and 31 with psoriatic arthritis. All patients were co-medicated with nonsteroidal anti inflammatory drugs (NSAID) as Diclofenacum, Indomethacinum, Piroxicamum and 51% with glycocorticosteroides. During the follow-up study we collected 18 case reports with ADR for 17% of the patients. From the patients with registered ADR, 11 was treated with standard dose of 7.5 mg MTX for a week and 7 patients received from 10 to 15 mg for a week. The distribution of the cases according patients' gender was 9 females and 7 males. Prevail individuals in age groups' 41 50 and over 61 years. The most frequent adverse drug reactions were leucopenia, trombocytopenia, skin reactions and gastrointestinal disorders as vomiting, melaena, epigastrial pain, etc. The primary risk connected with long therapy of low doses MTX is hepatotoxicity that diagnose and treatment are painful and expensive. As a result of the appearance of ADR in 5 patients the therapy with MTX was not changed, in two cases MTX is stopped timely or the dosage is changed and in the rest 11 patients MTX was excluded from the therapeutic scheme. PMID- 11059097 TI - Cytotoxicity test of Ipomoea involucrate. PMID- 11059098 TI - Synthesis of some N1-phenyl-N3-[2-aryl/aryloxymethyl-1,3,4-oxa(thia)diazol-2 yl]sulphonyl ureas as potential pesticides. AB - N1-Phenyl-N3-aryl/aryloxymethyl[1,3,4-oxa(thia)diazol-2-yl]s ulphonyl ureas (4 and 7) were conveniently prepared from 2-aryl/aryloxymethyl- 1,3,4 oxa(thia)diazol-2-sulphonamide (3 and 6) and phenyl isothiocyanate. Sulphonamides were synthesised by the reaction of 2-aryl/aryloxymethyl-1,3,4-oxa(thia)diazol-2 sulphonyl chlorides (2 and 5) with ammonia. All the compounds have been tested for fungicidal activity against the fungal species Cephalosporium saccharii and Helminthosporium oryzae. The two species of the aquatic fungi viz. Saprolegnia parasitica and Achlya orion responsible for fish mycoses were tested and also found remarkably active. All the compounds were also tested for herbicidal activity against Digiteria ciliaris, Oryza sativa and Raphunus sativa. PMID- 11059099 TI - Synthesis and antimicrobial activity of new 6-phenylimidazo[2,1-b]thiazole derivatives. AB - A series of new N2-substituted-6-phenylimidazo[2,1-b]thiazole-3-acetic acid hydrazides (2a-j) were synthesized and evaluated for in vitro antimicrobial activity against various bacteria and fungi. Some of the compounds showed antimicrobial activity against Staphylococcus aureus ATCC 6538, Staphylococcus epidermidis ATCC 12228, Trichophyton mentagrophytes var. Erinacei NCPF-375, Trichophyton rubrum and Microsporum audounii (MIC 25-0.24 micrograms/ml). The in vitro antimycobacterial activity of the new compounds was also investigated. The compounds exhibited different degrees of inhibition (17.98%) against Mycobacterium tuberculosis H37Rv in the primary screen that was conducted at 12.5 micrograms/ml using the BACTEC 460 radiometric system. PMID- 11059100 TI - Evaluation of Prosopis africana gum in the formulation of gels. AB - Prosopis africana gum was evaluated for use in the formulation of gels. The rate of release of salicylic acid from gels prepared from prosoopis gum was investigated. The rate of permeation of the drug through the gel was also evaluated. Surfactants were incorporated into the gels and the effect on the release and permeation was also investigated. Tragacanth gum gel was also prepared and used as the standard. The release and permeation of the drug from the gel was low. Incorporation of surfactants did not enhance the release of the drug. However the low release and permeation rates may be due to the poor water solubility of the incorporated drug. Correlation of the quantity of drug released with viscosity shows that drug release was dependent on the viscosity of the gels; the highly viscous gels showed slower release rates. PMID- 11059101 TI - Permeation kinetics of ketotifen fumarate alone and in combination with hydrophobic permeation enhancers through human cadaver epidermis. AB - Permeation kinetics for ketotifen fumarate from transdermal system through hairless mice skin and hog skin have been reported. However, permeation of ketotifen fumarate through human skin is not available till date. In view of the above, permeation profile and related kinetic parameters of ketotifen fumarate alone and in presence of three compatible enhancers namely Myristic acid isopropyl ester, Palmitic acid propyl ester and Lauric acid propyl ester through normal and delipidized human cadaver have been reported in the present study. The observed permeability flux, permeation coefficient and epidermal partition coefficient through 50% dried human cadaver epidermis were found to increase in presence of the three enhancers, amongst which the effect was found to be maximum with Myristic acid isopropyl ester. The more pronounced enhancing effect of Myristic acid isopropyl ester regarding permeability flux, permeation coefficient, epidermal partition coefficient and diffusion coefficient was attributed with solubility parameter being nearer to the skin lipid solubility parameter and probably due to its passage across the skin barrier through the lipid pathway. PMID- 11059102 TI - [Phase diagrams in the binary system]. AB - The properties of pharmaceutical formulations are profoundly influenced by physico-chemical interactions, particularly those occurring between a drug and an excipient. The prediction and comprehension of these interactions provide information to use during the development of a dosage form. Thermal analysis methods are commonly utilized to obtain data from which phase diagrams can be easily constructed for a number of binary systems. The aim of this review is the classification of the phase diagrams of binary systems, as a function of two fundamental factors: 1) the relative strength of interaction between similar (UAA, UBB) and different atoms (UAB); 2) the limiting permissible degree of deformation of the energy field. The main binary phase diagrams have been analyzed starting from the simplest case, in which the two components are virtually incapable of dissolving each other either in the liquid and in the solid states, to the case where the formation of unlimited solid solutions both in the liquid and in the solid states can be observed. PMID- 11059103 TI - Properties of films prepared from prosopis and mucuna gums. AB - Various properties of the films prepared from mucuna and prosopis gums have been evaluated. Sodium carboxymethylcellulose was used for comparison. When studied for elongation at break point, the most elastic of the gums was prosopis gum, followed by sodium carboxymethylcellulose, while mucuna gum was the least elastic. On microscopy, prosopis and sodium carboxymethylcellulose formed films of equal consistency without pores. PMID- 11059104 TI - Essential oil analysis of Eusteralis deccanensis Panigrahi (Lamiaceae). AB - The phytochemical potential of Eusteralis deccanensis has been revealed for the first time. The volatile oil obtained from the leaves and flowers was found to be rich in alpha- and beta-patchoulene. In addition, GC analysis revealed three sesquiterpenes (beta-caryophyllene, beta-bourbonene, beta-elemene), two oxygenated monoterpenes (geranyl acetate, neryl acetate) and one sesquiterpene alcohol (delta-cadinol). The plant possesses a patchoulene chemotype. PMID- 11059105 TI - Alterations in lymphocyte subpopulations in peripheral blood at manifestation of type 1 diabetes mellitus in childhood. AB - BACKGROUND: Alterations in cellular immunity at manifestation of type 1 diabetes mellitus, as described in publications so far, are equivocal. Moreover, the age of children was usually not taken into account. OBJECTIVES: Exact inferentially statistical measures were used to arrive at reliable information. METHODS: Thirty four diabetic children and 48 normals were taken randomly according to the established criteria, and scrutinized. Lymphocyte subpopulations counts were measured by flow cytometry using three-color-labelled monoclonal antibodies against cell surface markers. The resulting absolute cell counts as well as percentages from the total lymphocyte count were expressed in terms of univariate and bivariate 95% confidence intervals. They render an illustrative way for defining statistically significant (alpha = 5%) differences between health and disease. RESULTS: The CD8, CD16 absolute counts in younger diabetics were significantly decreased in average to 96-58% of the normal subgroup. For older children, CD4, CD8, CD16 and CD19 absolute counts were significantly lowered to 75-61% of the norm. Relative changes in Ly subpopulations were less pronounced. The immunoregulatory index increased significantly to 125-128% of the norm in either age group. The proportion of CD4 memory cells from the total of naive and memory cells was significantly increased to 122-133% of the norm in diabetic children of either age group. CONCLUSION: More significant changes of lymphocyte subpopulations than those given in literature were revealed at manifestation of childhood type 1 diabetes. They testify to the autoimmune pathogenesis of the type 1 diabetes mellitus. (Tab. 3, Fig. 4, Ref. 18.) PMID- 11059106 TI - [Virokines and viroceptors--viral immunomodulators with clinical and therapeutic implications]. AB - During evolution viruses have developed variety of sophisticated strategies for interactions with the immune system of the host. One of the defense strategies that counteract the immune responses of the infected organism exploits viral proteins that directly interfere with the host's cytokine system. Among such immunomodulatory molecules are classed also viral homologs of cytokines (virokines) and viral homologs of cytokine receptors (viroceptors), produced and secreted by the virus-infected cell. Virokines and viroceptors are encoded by large DNA viruses--herpesviruses and poxviruses. The respective genes have been obviously "stolen" by viruses from the host genomes and then have been modified. Detailed characterization of these viral elements, which induce or subvert the host's cytokine responses against viral infection, may contribute to a better understanding of the mechanisms which help the viruses to escape immune surveillance. Such knowledge have potential implications for viral epidemiology, treatment or prevention of viral and inflammatory diseases, and for the development of safer vaccines. Examples of viruses indicate, that "capturing" of the immunomodulatory genes may be a more general strategy used also by other types of pathogenic or parasitic organisms to evade the immune responses of their hosts. (Tab. 2, Fig. 2, Ref. 73.) PMID- 11059107 TI - [Circadian rhythm of the ventricular fibrillation threshold in a model of hypoventilation-reoxygenation in female Wistar rats]. AB - Hypoxia, similarly as myocardial ischemia, decreases the electric stability of the heart and thus produces the conditions for the genesis of ventricular arrhythmias within the 24 h period however a prompt restoration of oxygen supply or blood flow within myocardium causes serious ventricular arrhythmias. Therefore the aim of our study was to evaluate the reoxygenation impact on myocardium after hypoventilation in circadian dependence. The experiments were performed in female Wistar rats (pentobarbital anesthesia 40 mg/1 kg i.p., open chest experiments). The animals were adapted to the light regimen 12:12 hours, with the dark phase from 18.00 h to 06.00 h. Normal ventilation was used in the control group (n = 17) and in the second one (n = 4), 20 min reoxygenation followed 20 min of hypoventilation. The heart rate (HR) had been recorded just before the ventricular arrhythmias rose. Hypoventilation decreased significantly (p < 0.001) the VFT and HR when compared with the control group and changed the 24-hour rhythm of VFT to moderate bi-phase one. Reoxygenation counter changed this rhythm to inverse in the comparison with the control group. The lowest VFT values occurred when the top of VFT circadian rhythm was detected during normal ventilation. No dependence was detected between VFT and HR in both ventilation types. It is concluded that reoxygenation alternates the myocardial vulnerability to ventricular arrhythmias in dependence on alternation of light and dark and without the evident dependence on HR in the course of the whole 24-hour period. (Tab. 2, Fig. 2, Ref. 43.) PMID- 11059108 TI - [The vascular endothelium in inflammatory processes in vivo and in vitro]. AB - Endothelial injury occurs in atherosclerosis, infectious, rheumatic and vasculitic processes, leading to activation of transcription factors and endothelial expression of various cytokines and adhesion molecules. Endothelial cell cultures represent a valuable tool in research activities, with emphasis on the principal characteristics of angiogenesis, inflammatory response, transduction signals and endothelial functionality. In the laboratory of tissue cultures we prepared primary endothelial cultures by their isolation from the umbilical vein. This model system has been used to investigate the endothelial activation in vitro, adhesion alterations of immunocompetent cells to endothelium, adhesion molecule expression in the disease course monitoring and anti-inflammatory treatment. (Tab. 1, Fig. 5, Ref. 45.) PMID- 11059109 TI - [Comparison of various diagnostic kits and antigens for the detection of chlamydial antibodies in the serum of patients with respiratory diseases] . AB - Examination of sera of patients with respiratory diseases did not reveal any substantial difference in detection of chlamydial antibodies by ELISA using two commercial kits (Chlamydia STAT and rRLISA) demonstrating IgG antibodies and two corpuscular antigens prepared from Chlamydia psittaci and Chlamydia pneumoniae in the Institute of Virology, SAS, in Bratislava detecting whole serum antibodies. However, higher sensitivity of rELISA diagnostic kit was found when analyzing immunoglobulin classes, namely in case of IgG antibodies. Results of ELISA were in a great majority of cases, confirmed by examination of sera by microimmunofluorescence test, corpuscular antigens from the Institute of Virology being somewhat more sensitive than the commercial Micro-IF test kit. (Tab. 4, Ref. 23.) PMID- 11059110 TI - [Tularemia--an old and a new problem in the South Moravia Region]. AB - BACKGROUND: The first epidemic of ulceroglandular forms of tularemia acquired in coincidence with the manipulation with tularaemic hares took place in 1936 in the surroundings of Breclav and Valtice. The largest epidemic took place in the 1960's when hundreds of agricultural workers in the initial phases of the production of sugar within sugar refineries were afflicted by pulmonary forms of this disease. In the subsequent period which was interrupted only by smaller local epidemics, the number of new cases were gradually decreasing to the minimum at the beginning of 1990's. However, since 1994, the number of cases has began to increase again, namely those afflicted by ulceroglandular and oroglandular forms. OBJECTIVES [corrected]: In consequence of the long absence of this disease in clinical practice, the diagnostic awareness has decreased, and therefore the author has decided to indicate and review the current basic data on epidemiology and clinical manifestations of tularaemia. GROUP OF PATIENTS AND METHODS: The author has analysed the documentation of 577 of adults afflicted by tularemia and medically treated at the clinic of the Faculty Hospital in Brno in the period form 1959 to 1999. The study reviews the onset of the disease and the pathway of transmission of infection and its clinical manifestation. MAIN RESULTS: Following the long-termed sporadic occurrence of tularaemia after major epidemics of pulmonary forms of this disease in 1960's, interrupted only by smaller local epidemics, the incidence has began to increase again in 1994. The number of pulmonary forms has decreased whereas the occurrence of ulcerulceroglandular and oroglandular forms has increased. Hares have become the source of infection again. CONCLUSIONS: The fact that tularemia has repeatedly become a threat in Southern Moravia should be taken into account in the assessment of diagnosis in cases with unclear lymphadenitis and febrile states which defy the penicillin treatment especially in winter. (Tab. 3, Fig. 3, Ref. 23.) PMID- 11059111 TI - [Frequency analysis of the QRS complex: a new method of detecting development of myocardial damage after anthracycline cytostatics]. AB - The aim of the study was to investigate the effect of the new modification of frequency-domain analysis of ECG signal (gliding-window FFT) in long-term monitoring of cardiac status of patients who received a potentially cardiotoxic anthracycline therapy in childhood. Area ratio (60-120/0-120 Hz) peaks within QRS complex were significantly higher in 60 patients compared with 70 healthy children and young adults. Persistent abnormalities in frequency content of ECG signal in oncologic patients might indicate myocardial damage induced by anthracyclines. (Fig. 1, Ref. 17.) PMID- 11059112 TI - [Professionalism of teachers of theoretical and clinical disciplines in medical schools]. AB - Authors submit their view on the possibility of performing a serious evaluation of professionalism of teachers of theoretical and clinical subjects, particularly at school of medicine. The authors compare their ideas with those of A.C. Orstein and D.U. Levine (1989). In conclusion they point out the extraordinary position of university teachers so-called clinicians in the system of pregradual and post gradual education, and their evaluation. (Fig. 1, Ref. 15.) PMID- 11059113 TI - Acculturation of South Asian adolescents in Australia. AB - BACKGROUND: This investigation is part of a wider research project on the acculturation and identity-related issues of South Asian young people in Britain, Canada and the US. There has been no published research on the acculturation of this ethnic group in Australia. The project is embedded within Berry's (1994) theoretical framework of acculturation. AIMS: There are two major aims of the research. Firstly, to discover any pattern of acculturation of South Asian young people and to compare it with that of their counterparts in Britain and Canada. Secondly, to validate the existing Acculturation Scale (Ghuman, 1975, 1997, 1999a) with a group of South Asian young people living in a different socio political context. SAMPLES: The sample was drawn from three high schools--two being in Newcastle City, and one in a rural setting of New South Wales, Australia. A representative sample of 75 boys and girls (aged 14 to 16) from two social class backgrounds took part in the research. METHODS: The young people were requested to fill in a background questionnaire on their religion, languages spoken at home, best friend, frequency of visit to a temple, etc. They also completed the Aberystwyth Likert-type Acculturation Scale. RESULTS: Summated scores on the scale of two social class groups confirmed the previous finding that a non-manual group shows a higher degree of acculturation compared with a manual group. But, contrary to expectations, girls show lower acculturation than boys. The Spearman-Brown coefficient of reliability is 0.82--which is in line with the findings in England and Canada. CONCLUSIONS: The whole sample shows a lower degree of acculturation compared with their counterparts in Canada and England. This is fully contextualised: viz, at the time of research (March 1998) the socio-political of Australia was anti-Asian, due mainly to the impending general election in which immigration from Asia was a significant issue. The mean score of the sample reflects their bilingualism and biculturalism. PMID- 11059114 TI - Developing written discourse knowledge in whole language and code emphasis classrooms. AB - BACKGROUND: Although there have been a plethora of studies comparing the effects of whole language vs. code emphasis instruction on children's literacy development, few have examined what children actually learn about features of written discourse in the two instructional settings. AIM: The purpose of this study was to compare the development of written discourse knowledge among young children in whole language and code emphasis classrooms. SAMPLES: Participants were 64 first grade children, 29 boys and 35 girls, in four intact classrooms from a mid-western school district of the United States. About half of them (N = 34) received whole language instruction and the other half (N = 30) code emphasis instruction. METHODS: Each child was individually asked to compose a book-like story about a personally relevant experience for others to read at the beginning and end of the school year. The texts were analysed linguistically and statistically in terms of three fundamental features of written discourse- autonomy, conventionality, and specialised grammar. RESULTS: The children developed more knowledge about the autonomy and conventionality features of written discourse, but their understanding of its specialised grammar remained inchoate. Further, the whole language and code emphasis groups did not demonstrate statistically significant differences in their working knowledge of written discourse. CONCLUSIONS: The nature of instructional programme (whole language vs. code emphasis) appears to have little impact on children's developing understanding of written discourse. PMID- 11059115 TI - Effects of different forms of school contact on children's attitudes toward disabled and non-disabled peers. AB - BACKGROUND: There have been fluctuations in research interest into the inclusion of children with disabilities in mainstream schools over the last twenty years. It is still not clear what methods, practices and types of contact are most likely to promote positive attitudes in children toward disabled peers and disability generally. AIMS: To consider two theoretical models of inter-group contact, both claiming to identify precursors for generalised attitude change, in relation to the attitudes of non-disabled children toward disabled peers as a function of different classroom contact. SAMPLE: Participants were 256 non disabled school children aged 5-11 years (128 girls and 128 boys). METHODS: Measures of sociometric preference and the evaluation of psychological and physical attributes were used to ascertain children's perceptions of known and unknown peers with disabilities. RESULTS: A relationship was found between the type of contact the children had with disabled peers, and their perceptions of psychological and physical attributes (stereotypes) of groups of unknown disabled and non-disabled peers. CONCLUSIONS: Results show generalisation of stereotypic attitude/judgments from one type of disability to another as a consequence of the two types of contact situation. Findings have important implications for integrating disabled children into mainstream. PMID- 11059116 TI - The effects of grades on course enjoyment: did you get the grade you wanted? AB - BACKGROUND: Students tend to rate university courses more positively if they do well. Greenwald and Gillmore (1997a) suggested that it is not students' absolute grades that are important but rather how these grades compare to their expectations. However, this hypothesis is difficult to evaluate because few studies have measured grade expectations at the beginning of courses. AIM: By measuring students' grade expectations and enjoyment at several stages during a course, we hoped to evaluate the extent to which expectations modulate the impact of grades on course enjoyment. SAMPLE: Participants were 242 students in a university course in psychology. METHOD: Students were asked what grades they expected, and how much they were enjoying the course, at four stages. The effect of grades and grade expectations on enjoyment were analysed using restricted maximum likelihood (REML) and regression analyses. RESULTS: The best predictor of course enjoyment varied somewhat at different stages, but in general it was the extent to which students' grades surpassed their expectations. Students' expectations at the beginning of the course proved particularly influential. CONCLUSIONS: Grade expectations do influence how students react to course grades, but the prominent role of pre-course expectations suggests that it may be important to distinguish between grade aspirations and grade expectations. It appears to be students' aspirations--the grades they hope to achieve--that most strongly shape their emotional reactions, rather than the more realistic expectations they may form later in a course. PMID- 11059117 TI - (Re-) constructing pre-linguistic interpersonal processes to promote language development in young children with deviant or delayed communication skills. AB - BACKGROUND: Approaches to intervention with young children with delayed or deviant communication skills have tended to focus directly on the production and comprehension of linguistic forms. Research in developmental psychology, however, has highlighted the role of joint attention and action within a mutually negotiated frame in early communicative development. AIMS: An approach to intervention was developed which uses graded participation in such 'formats' to promote language development in 3- to 5-year-olds with significant delays or deviance in communication skills. This study was designed to evaluate this in comparison with a more conventional approach. SAMPLE: The sample comprised twenty 3- to 5-year-olds attending a diagnostic nursery or a speech and language support centre attached to a mainstream primary school. METHOD: In the intervention, developmentally ordered sequences of exchange games provided predictable ruled based and structured joint contexts of action, within which verbal and non-verbal exchanges between an adult and child could be meaningfully used. In the control condition, participants received more conventional intervention focusing more directly on language per se. Level of participation in dyadic play was assessed in terms both of role and cognitive complexity, and for the present study language development was assessed using the verbal subtests of the WPPSI. The study entailed both a between- and a within-subjects design. RESULTS: Cross sectional and longitudinal comparisons confirmed that this form of intervention was significantly more effective than the control condition in increasing children's level of participation in social game formats and in promoting their language development; moreover, these were significantly correlated. CONCLUSIONS: The results indicate that the development of language skills can be promoted more effectively through (re-) constructing the interpersonal framework of pre linguistic communicative development than through conventional intervention focusing directly upon language per se. PMID- 11059118 TI - Children's perceptions of school-based violence. AB - BACKGROUND: An important first step in understanding school-based violence is understanding children's subjective perceptions of the phenomena. Understanding these perceptions is likely to be a major factor in determining the integrity of both prevalence and intervention studies. AIMS: We investigated how elementary and secondary aged children perceived school-based violence. SAMPLE: A sample of 979 children from a nested random sample of elementary (grades 3-6) and middle school (grades 7-8) classrooms in Jerusalem participated in this study. METHODS: To understand children's perception of school violence, we used an instrument composed of 19 dichotomous items, each presenting a one-line description of a behaviour, which the respondent would define as either 'intentionally harmful' or not. RESULTS: Eighth graders were significantly less likely to label the behaviours described as violent compared to all other grades; and seventh graders were less likely as compared to third, fourth and fifth graders; also, some between-gender differences were found. CONCLUSION: The respondents often view the behaviours described as intentional and aggressive; this finding should serve as an impetus to widen the scope of school-based violence interventions to include these behaviours, especially for younger children. PMID- 11059119 TI - Item-level and subscale-level factoring of Biggs' Learning Process Questionnaire (LPQ) in a mainland Chinese sample. AB - BACKGROUND: The learning process questionnaire (LPQ) has been the source of intensive cross-cultural study. However, an item-level factor analysis of all the LPQ items simultaneously has never been reported. Rather, items within each subscale have been factor analysed to establish subscale unidimensionality and justify the use of composite subscale scores. AIMS: It was of major interest to see if the six logically constructed items groups of the LPQ would be supported by empirical evidence. Additionally, it was of interest to compare the consistency of the reliability and correlational structure of the LPQ subscales in our study with those of previous cross-cultural studies. METHODS: Confirmatory factor analysis was used to fit the six-factor item level model and to fit five representative subscale level factor models. SAMPLE: A total of 1070 students between the ages of 15 to 18 years was drawn from a representative selection of 29 classes from within 15 secondary schools in Guangzhou, China. Males and females were almost equally represented. RESULTS: The six-factor item level model of the LPQ seemed to fit reasonably well, thus supporting the six dimensional structure of the LPQ and justifying the use of composite subscale scores for each LPQ dimension. However, the reliability of many of these subscales was low. Furthermore, only two subscale-level factor models showed marginally acceptable fit. Substantive considerations supported an oblique three-factor model. CONCLUSIONS: Because the LPQ subscales often show low internal consistency reliability, experimental and correlational studies that have used these subscales as dependent measures have been disappointing. It is suggested that some LPQ items should be revised and other items added to improve the inventory's overall psychometric properties. PMID- 11059120 TI - Parents' education, cognitive ability, educational expectations and educational attainment: interactive effects. AB - BACKGROUND: The models that have been used so far to describe the process underlying educational attainment have been almost always linear. Little research has been aimed at studying interactions among the determinants of educational attainment. AIM: The aim of the study is to examine the interactions between parents' education, cognitive ability and educational expectations in determining educational attainment. SAMPLE: Participants were 8570 Americans who were born between 1957 and 1964. METHOD: The information was taken from the National Longitudinal Survey of Youth. Information about parents' education, cognitive ability and educational expectations was taken from the 1979 survey. Information about educational attainment was taken from the 1991 survey. RESULTS: The findings indicate that there is an offsetting relationship between the education of the two parents in the formation of expectations, but not in the determination of attainment; and that, both for expectations and for attainment, the cognitive ability of the child has an offsetting relationship with mother's education but not with father's education. The findings also indicate that there is a synergistic relationship between cognitive ability and educational expectations in determining educational attainment. CONCLUSIONS: There are theoretically meaningful interactions between the determinants of educational attainment. The pattern of these interactions capture some of the intricate psychological processes underlying the combined influence of background variables and children's characteristics on educational attainment. PMID- 11059121 TI - Social competition in school: relationships with bullying, Machiavellianism and personality. AB - BACKGROUND: Bullying is investigated as part of the individual's general framework of attitudes towards interpersonal relationships, social competition and motivation in school. AIMS: It was hypothesised that bullying behaviour and pro-bullying attitudes would be associated with socially competitive attitudes in the classroom, Machiavellianism, and the personality constructs of Psychoticism and Extraversion. SAMPLES: 198 9- to 12-year-old children from two Glasgow primary schools. METHODS: Children completed several measures: a newly developed questionnaire assessing motivations behind social competition and effort in class, the Kiddie-Mach scale, the Pro-Victim scale, items from the Olweus Bullying Questionnaire, and the Junior Eysenck Personality Questionnaire. RESULTS: A 'Desire for social success' factor (incorporating a deliberate lack of effort) was negatively correlated with support for victims of bullying, even after partialling out Machiavellianism, Psychoticism, and social desirability. Pro-victim attitudes were in turn negatively correlated with Machiavellianism and Psychoticism, and positively correlated with Lie score. Finally, children categorised as bullies scored significantly higher than controls on Machiavellianism, and significantly lower in terms of pro-victim attitudes. CONCLUSIONS: Results are discussed in terms of further study and implications for classroom practice and anti-bullying policy. PMID- 11059122 TI - Hepatitis C: medical information update. Canadian Liver Foundation. National Hepatitis C Education Program. PMID- 11059123 TI - Current status of hepatitis C in Canada. PMID- 11059124 TI - Hepatitis C in Canada's first nations and Inuit populations: an unknown burden. PMID- 11059125 TI - Public health and hepatitis C. AB - This paper reviews key public health aspects related to surveillance, transmission and primary prevention of hepatitis C. Hepatitis C is now a reportable disease in all Canadian provinces and territories. Although prevalence in Canada is estimated at under 1%, that associated with injection drug use (IDU) approaches 90%. The epidemiology of new HCV infections in Canada is now primarily defined by IDU behaviour, with annual incidence rates among new drug injectors exceeding 25%. HCV is less efficiently transmitted through other routes of exposure. An effective vaccine against HCV remains elusive. Some jurisdictions offer hepatitis A and hepatitis B vaccine to HCV-infected persons. An array of harm reduction strategies targeting IDU has been implemented but underdeployed across Canada, and has been ineffective to date in controlling the HCV epidemic. Public policy alternatives, such as legalization and regulation of injection drugs, are being debated. Improved HCV preventive strategies are urgently required and need careful evaluation. PMID- 11059127 TI - The hepatitis C prevention, support and research program: Health Canada initiatives on hepatitis C. Health Canada. AB - In September 1998, Health Minister Allan Rock announced new federal hepatitis C funding of +50 million over five years for initiatives relating to community based support, research, and disease prevention. Since then, broad cross-country consultations have taken place with individuals and their caregivers who are infected, or affected, by this disease; non-governmental organizations; provinces and territories; and health care professionals. The result is a relevant, compassionate and targeted new Health Canada Hepatitis C Program with a four point action agenda, that encompasses five components--prevention; community based support; care and treatment support; research; and ongoing management, evaluation and public involvement. PMID- 11059126 TI - Treatment options in patients with chronic hepatitis C. AB - Hepatitis C is a major health care problem plagued by the lack of a truly effective therapy. To date, the combination of interferon and ribavirin has provided the best chance of viral eradication. However, this therapy is expensive, has multiple side effects and works in less than half of patients. New strategies need to be developed to deal with the increasing burden of hepatitis C related disease, and we anxiously await the arrival of new drugs such as helicase and protease inhibitors. PMID- 11059128 TI - Living with hepatitis C. PMID- 11059129 TI - Through the eyes of a mother. PMID- 11059130 TI - Living with hepatitis C as a nurse. PMID- 11059131 TI - Hepatitis C virus diagnosis and testing. AB - Development of serological and nucleic acid testing (NAT) has revolutionized hepatitis C virus (HCV) diagnosis. Although third generation anti-HCV enzyme immunoassays (EIAs) are very effective for testing high prevalence populations, confirmatory testing is still necessary when these tests are applied to populations with a low HCV prevalence to exclude false positive results. Limitations of third generation anti-HCV EIAs include: the relatively prolonged time between acute infection and detection of seroconversion (which typically requires at least 5-6 weeks); delayed seroconversions in immunocompromised hosts (requiring months to years); and the inability of serological tests to confirm active HCV infection. In contrast, nucleic acid testing (NAT) can directly detect HCV RNA in serum, plasma or tissue and thereby confirm active infection as well as narrow the window between infection and HCV detection to as little as 1-2 weeks. Commercial NAT assays are now highly sensitive, specific, and reproducible and have largely replaced unreliable home brew nucleic acid amplification assays. Qualitative commercial NAT are typically more sensitive than quantitative assays and therefore the method of choice to confirm active infection. Given the efficacy of combination therapy with interferon/ribavirin and newer antiviral agents under development, HCV infection may become curable, which will likely impact future disease transmission. As the therapeutic costs are currently very high, there is clearly a need to assess the utility of quantitative NAT and to further evaluate the role of HCV genotyping to optimize antiviral therapy. Thus for the foreseeable future, a combination of both serological tests and NAT will be required for cost-effective HCV diagnosis and monitoring. PMID- 11059132 TI - Building a better blood system for Canadians. PMID- 11059133 TI - Hepatitis C: mental health issues. PMID- 11059134 TI - [Pars plana vitrectomy of subretinal neovascular membranes in age-related macular degeneration]. AB - The possibilities of treatment of chorioideal neovascular membrane (CNV) in age related macular degeneration are very limited. Conservative treatment has no effect, laser coagulation is accepted nowadays as the method of choice only in a selected group of patients and its results are more than disappointing. Surgical extraction of subretinal neovascular membranes opens wider opportunities for improvement of optic functions in indicated patients, in the majority it makes their stabilization possible. The authors present a group of their own 20 patients where extraction of CNV was performed. They describe the technique of extraction, indications and results. PMID- 11059135 TI - [Removal of the juxtafoveal neovascular membrane in age-related macular degeneration]. AB - Surgical extraction of the submascular neovascular membrane (SNM) is one of the new indications for surgery of the macula. The functional results after surgical extraction of the SNM depend on the state of the submascular pigmented epithelium and neuroepithelium which is, last not least, conditioned by the etiology of SNM. The majority of SNM in age-related macular degenerations has a poor prognosis, These SNM are as a rule nourished from several vascular sources and markedly destroy the pigmented epithelium because they penetrate beneath and above it. The authors evaluate in the submitted paper the results of surgical removal of the SNM in two eyes of two patients with age-related macular degeneration. Despite the relatively favourable juxtafoveal localization of the membrane, a non complicated course of the operation and postoperative course the functional result of the operation in the two eyes was not very satisfactory. Chorioretinal scars with extensive defects of the pigmented epithelium in the region of the macula were the cause of a decline of visual acuity to a level which however could be hardly preserved by a conservative procedure. PMID- 11059136 TI - [Ultrastructural analysis of tissue removed during surgery of idiopathic macular holes]. AB - AIM: To analyze the ultrastructural features of epimacular tissue removed during idiopathic macular hole (IMH) surgery and to compare them with those of idiopathic epimacular membranes. METHODS: Three consecutive patients with unilateral stage 3 IMH underwent pars plana vitrectomy with surgical removal of delicate gelatinous membranous epimacular tissue. One patient was previously treated with laser photocoagulation. The excised material was investigated by means of light and transmission electron microscopy. Results were compared with the findings of the excised material in two idiopathic epimacular membranes. RESULTS: The excised material included variable proportions of cellular and membranous material. Glial cells (Muller cells, fibrous astrocytes) and fibroblast-like cells dominated in the cellular component both in the specimens from IHM eyes and in the comparative material. Neural elements were revealed in two of three specimens from IMH eyes. Membraneous material was formed by fragments of internal limiting membrane. CONCLUSIONS: Dominance of the glial cells in the cellular component of epimacular formations may reflect the important role of these cells both in formation and in healing process of IMH. The presence of neural elements in the removed epimacular tissue suggests that, at least in some cases, IMH surgery with peeling of an epimacular membrane can be associated with some injury of the macula. PMID- 11059137 TI - [Diagnosis of swollen optic disks using color Doppler ultrasonography] . AB - The Goal of this study was prospective evaluation of the blood flow parameters maximal systolic velocity (MSV), minimal diastolic velocity (MDV) and Pourcelot's resistivity index (RI) in patients with swollen optic dis. 63 patients (79 eyes) were examined with swollen optic disc-10 patients (17 eyes) with intracranial hypertension, 12 patients (14 eyes) with pseudoedema of disk of the optic nerve, 7 patients (7 eyes) with inflammation swollen optic disc, 3 patients (6 eyes) with pseudotumor cerebri, 8 patients (12 eyes) with arteritic anterior ischemic optic neuropathy (A-AION) and 23 patients (23 eyes) with nonarteritic anterior ischemic optic neuropathy (N-AION) and 23 patients (23 eyes) with nonarteritic anterior ischemic optic neuropathy (N-AION). All patients had ophthalmological examinations and Color Doppler ultrasonography in central retinal artery (CRA), posterior ciliary artery (PCA), ophthalmic artery (OA) and in central retinal vein (CRV). The blood flow parameters of the swollen optic disc and normal optic disc were not significant changed at intracranial hypertension, pseudooedema cerebri, inflammation swelling, and at pseudotumour od the optic disc. A-AION and N-AION acallocated significant changes of the blood velocities and resistivity index. A-AION: significant increasing resistivity index at CRA, CPA, significant decrease of MSV and MDV and difficult mapping of the CPA in Color doppler mapping. At the fellow eye (without swollen optic disc) was significant decreasing of the blood velocities of the CRA, CPA, but not so much as at the defective eye with swollen optic nerve disk. N-AION: defective eye with swollen optic disc--there were: nonsignificant decreasing of MSV, MDV, significant decreasing of MSV, MDV and increasing of resistivity index of the CRA, CPA at the fellow eye. AO was without significant changes. Color Doppler information allowed to specify diagnosis of A-AION and N-AION. PMID- 11059138 TI - [Intraocular lens implantation in children]. AB - OBJECTIVE: To evaluate the results of primary and secondary implantations of intraocular lenses (IOL) in children--visual acuity, binocular function, resulting refraction and complications after operation. METHODS: The group comprises 47 eyes of 36 children operated in 1993-1997, aged 5-16 years (mean age of operated children 10.3 years). The follow up period after operation was 18-60 months. The mean follow period was 37 months. Twenty-two primary implantations of posterior chamber IOL were made and 25 secondary implantations, incl. 13 posterior chamber IOL and 12 anterior chamber IOL. From the total number of cataracts 26 were traumatic and 21 congenital (16 bilateral and 5 unilateral). RESULTS: A visual acuity of 6/12 or better was recorded after operation in 31 (67%) eyes, incl. vision 6/6 in 7 eyes. Vision 6/15 to 6/24 was found in 8 (18%) eyes. Binocular function after operation was recorded in 26 (73%) children. Of these 11 have stereopsis and 15 have fusion I-III. Nine children (27%) do not have binocular functions. Half the eyes have after operation a refraction within the interval +/- 1.0 D SE. Fifteen eyes (33%) are within the range of +/- 2 D SE from emetropy. Eight eyes had a refraction in the interval of +/- 3 D SE during the last check-up examination. The most frequent postoperative complication was a slight (48%) or more serious (23%) inflammatory reaction, decentration of the IOL (23%), and in case posterior capsulotomy was not performed during primary implantation, the complication in almost 8% of eyes was secondary cataract which had to be treated by capsulotomy with a NdYAG laser. One intraocular lens had to be explanted. CONCLUSION: Implantation of an intraocular lens in children aged 5 16 years is according to the results of postoperative visual acuity and binocular functions a suitable alternative of correction of child aphakia. PMID- 11059139 TI - [Cerebrospinal fluid cytology in retinoblastoma--possibility of early detection of tumor dissemination into the CNS]. AB - Retinoblastoma is a malignant tumourous which occurs in childhood. The most important factors which influence possible cure of patients with a retinoblastoma are early detection of the disease, correct diagnosis and adequate therapy. In the submitted case-record the authors provide evidence of the importance of cytological examination of cerebrospinal fluid which is important in retinoblastoma in particular for the follow-up of treatment. Cytological examination of cerebrospinal fluid in this type of tumour is very reliable and can reveal infiltration of tumour cells into the CNS much sooner than imaging methods. PMID- 11059140 TI - [Laser in situ keratomileusis in the treatment of myopia (retrospective study 1995-1996]. AB - In this article, the authors present their findings concerning the usage of the LASIK method in 1,602 cases of myopia. The LASIK procedure is shown to be a very effective and safe method in a high percentage of cases. All operations were performed under topical anesthesia with Oxybuprocain 0.4% and Lidocain 2.0%. Uncorrected visual acuity is used as the main criterion in assessing the effectiveness of refractive procedures, and our average values of improvement were from 0.08 pre-operatively to 1.01 post-operatively. The average spherical equivalent was improved from a pre-operative value of -5.74 D to a post-operative one of 0.26 D. Emetropy was achieved in 89% of cases, 8.5% of cases were undercorrected and 2.5% overcorrected. PMID- 11059141 TI - [Anterior chronic uveitis in chronic juvenile arthritis]. AB - BACKGROUND: Anterior uveitis (iridocyclitis) in association with juvenile chronic arthritis has two different clinical units: the acute and chronic form. They differ as to the clinical picture, course, complications, relation to antinuclear antibodies and HLA B 27 antigen, response to treatment and prognosis of optic functions. OBJECTIVE: To investigate the development of complications in patients with the chronic form of juvenile chronic arthritis in relation to the duration of uveitis. To focus attention on the treatment and prognosis of the disease. PATIENTS: The authors evaluated retrospectively 18 patients with anterior chronic uveitis and diagnosed juvenile chronic arthritis. The group comprised 13 girls and 5 boys, mean age 14.3 years (6-25 years) with a mean duration of uveitis of 7.5 years (2-21 years). RESULTS: Complications and deterioration of vision are frequent and serious in patients with the chronic form of uveitis. With the duration of the disease progression of complications occurs, their number increases and vision deteriorates. The adverse prognosis in some patients is promoted by an inadequate response to treatment. Six patients (8 eyes) were subjected to surgical treatment of complications. The postoperative results are not encouraging. CONCLUSION: The authors wish to draw attention to the need of close collaboration with a rheumatologist, paediatrician, ophthalmologist and the necessity to devote special care to children with juvenile chronic arthritis and uveitis. PMID- 11059142 TI - [The Hallgren syndrome]. AB - Hallgren syndrome is a hereditary disease with autosomal recessive inheritance. Its exact genetic background has not been elucidated so far. From the clinical aspect is comprises association of retinitis pigmentosa, atrophy of the optic nerve, nystagmus and congenital hearing damage combined with neurological and psychiatric symptoms. The authors describe two siblings with the clinical picture of this syndrome. It is a finding not published so far in the Czech literature. PMID- 11059143 TI - [Iridoschisis--case report]. AB - The authors present an account of a 78-year-old female patient with bilateral iridoschisis. The disease was present in the lower portions of the iris. The pupil was centred, mobile. Part of the ruptured fibres of the iris floated freely in the anterior chamber. The cornea was not decompensated. The patient had open angle glaucoma and cataract. The method of choice in the treatment of the condition is laser iridotomy or microsurgical iridectomy and extraction of the cataract with implantation of PC IOL. Conservative treatment attempts to prevent possible damage of the optic nerve. PMID- 11059144 TI - [Alternative method of reconstruction in traumatic loss of the upper eyelid]. AB - The authors wish to present an alternative technique of more than half upper lid defect reconstruction. In a situation, where the standard techniques couldn't be used due to the specific nature of the local findings, the described method seems to be a sound alternative for the lid reconstruction. PMID- 11059145 TI - [Evaluation of the quality of life]. PMID- 11059146 TI - [Modern trends in ophthalmologic genetics. I. Advances in molecular genetics in ophthalmology]. PMID- 11059147 TI - [To the 25th anniversary of the journal. Turning over the pages]. PMID- 11059148 TI - [Study of the brain organization of creativity. Part II. Positron emission tomography]. PMID- 11059149 TI - [The role of cerebral regulatory structures in formation of the human EEG]. PMID- 11059150 TI - [Functional organization of the cortex of large hemispheres during voluntary movements. Age factors]. PMID- 11059151 TI - [Perception of spatial relations between objects in early ontogenesis]. PMID- 11059152 TI - [Estimation of short musical fragments by normal subjects and patients with persistent impairment of depressive nature]. PMID- 11059153 TI - [Comprehensive bioelectrical analysis of mechanisms of the alternative state of consciousness]. PMID- 11059154 TI - [Preference of the high probability of attaining the goal or its subjective value]. PMID- 11059155 TI - [Evoked autonomic skin potentials (modern concepts of their mechanisms]. PMID- 11059156 TI - [Calcium mechanism and its regulation in man during adaptation to microgravity]. PMID- 11059157 TI - [Effect of the long-term space flight on the extracellular fluid volume]. PMID- 11059158 TI - [Information stress problem in operators]. PMID- 11059159 TI - [Comprehensive study of the ecological awareness in high school students]. PMID- 11059160 TI - [Autonomic regulation and mental activity in children during school studies under unfavorable climatic conditions of the middle Ob river]. PMID- 11059161 TI - [Aging prevention: the new science outline]. PMID- 11059162 TI - [The role of the spatial-frequency analysis, primitives, and the interhemispheric asymmetry in identification of visual images]. PMID- 11059163 TI - [Self-regulation of the autonomic homeostasis in emotional stress]. PMID- 11059164 TI - A brain-controlled switch for asynchronous control applications. AB - Asynchronous control applications are an important class of application that has not received much attention from the brain-computer interface (BCI) community. This work provides a design for an asynchronous BCI switch and performs the first extensive evaluation of an asynchronous device in attentive, spontaneous electroencephalographic (EEG). The switch design [named the low-frequency asynchronous switch design (LF-ASD)] is based on a new feature set related to imaginary movements in the 1-4 Hz frequency range. This new feature set was identified from a unique analysis of EEG using a bi-scale wavelet. Offline evaluations of a prototype switch demonstrated hit (true positive) rates in the range of 38%-81% with corresponding false positive rates in the range of 0.3% 11.6%. The performance of the LF-ASD was contrasted with two other ASDs: one based on mu-power features and another based on the outlier processing method (OPM) algorithm. The minimum mean error rates for the LF-ASD were shown to be significantly lower than either of these other two switch designs. PMID- 11059165 TI - ECG signal compression using analysis by synthesis coding. AB - In this paper, an elecrocardiogram (ECG) compression algorithm, called analysis by synthesis ECG compressor (ASEC), is introduced. The ASEC algorithm is based on analysis by synthesis coding, and consists of a beat codebook, long and short term predictors, and an adaptive residual quantizer. The compression algorithm uses a defined distortion measure in order to efficiently encode every heartbeat, with minimum bit rate, while maintaining a predetermined distortion level. The compression algorithm was implemented and tested with both the percentage rms difference (PRD) measure and the recently introduced weighted diagnostic distortion (WDD) measure. The compression algorithm has been evaluated with the MIT-BIH Arrhythmia Database. A mean compression rate of approximately 100 bits/s (compression ratio of about 30:1) has been achieved with a good reconstructed signal quality (WDD below 4% and PRD below 8%). The ASEC was compared with several well-known ECG compression algorithms and was found to be superior at all tested bit rates. A mean opinion score (MOS) test was also applied. The testers were three independent expert cardiologists. As in the quantitative test, the proposed compression algorithm was found to be superior to the other tested compression algorithms. PMID- 11059166 TI - A two-stage discrimination of cardiac arrhythmias using a total least squares based prony modeling algorithm. AB - In this paper, we describe a new approach for the discrimination among ventricular fibrillation (VF), ventricular tachycardia (VT) and superventricular tachycardia (SVT) developed using a total least squares (TLS)-based Prony modeling algorithm. Two features, dubbed energy fractional factor (EFF) and predominant frequency (PF), are both derived from the TLS-based Prony model. In general, EFF is adopted for discriminating SVT from ventricular tachyarrhythmias (i.e., VF and VT) first, and PF is then used for further separation of VF and VT. Overall classification is achieved by performing a two-stage process to the indicators defined by EFF and PF values, respectively. Tests conducted using 91 episodes drawn from the MIT-BIH database produced optimal predictive accuracy of (SVT, VF, VT) = (95.24%, 96.00%, 97.78%). A data decimation process is also introduced in the novel method to enhance the computational efficiency, resulting in a significant reduction in the time required for generating the feature values. PMID- 11059167 TI - Nonlinear transient chirp signal modeling of the aortic and pulmonary components of the second heart sound. AB - This paper describes a new approach based on the time-frequency representation of transient nonlinear chirp signals for modeling the aortic (A2) and the pulmonary (P2) components of the second heart sound (S2). It is demonstrated that each component is a narrow-band signal with decreasing instantaneous frequency defined by its instantaneous amplitude and its instantaneous phase. Each component is also a polynomial phase signal, the instantaneous phase of which can be accurately represented by a polynomial having an order of thirty. A dechirping approach is used to obtain the instantaneous amplitude of each component while reducing the effect of the background noise. The analysis-synthesis procedure is applied to 32 isolated A2 and 32 isolated P2 components recorded in four pigs with pulmonary hypertension. The mean +/- standard deviation of the normalized root-mean-squared error (NRMSE) and the correlation coefficient (rho) between the original and the synthesized signal components were: NRMSE = 2.1 +/- 0.3% and rho = 0.97 +/- 0.02 for A2 and NRMSE = 2.52 +/- 0.5% and rho = 0.96 +/- 0.02 for P2. These results confirm that each component can be modeled as mono-component nonlinear chirp signals of short duration with energy distributions concentrated along its decreasing instantaneous frequency. PMID- 11059168 TI - Improving the accuracy of the boundary element method by the use of second-order interpolation functions. AB - The boundary element method (BEM) is a widely used method to solve biomedical electromagnetic volume conduction problems. The commonly used formulation of this method uses constant interpolation functions for the potential and flat triangular surface elements. Linear interpolation for the potential on a flat triangular mesh turned out to yield a better accuracy. In this paper, we introduce quadratic interpolation functions for the potential and quadratically curved surface elements, resulting from second-order spatial interpolation. Theoretically, this results in an accuracy that is inversely proportional to the third power of element size. The method is tested on a four concentric sphere geometry, representative for electroencephalogram modeling, and compared to previous solutions of this problem in literature. In addition, a cylindrical test configuration is used. We conclude that the use of quadratic interpolation functions for the potential and of quadratically curved surface elements in BEM results in a significant increase in accuracy and in some cases even a reduction of the computation time with the same number of nodes involved in the calculations. PMID- 11059169 TI - Current multipole expansion to estimate lateral extent of neuronal activity: a theoretical analysis. AB - High-resolution magnetoencephalography (MEG) allows for a detailed description of focal neuronal current sources going far beyond the dipole approximation which merely indicates the center and magnitude of neuronal activity. Higher order multipole coefficients can be related to other bulk properties, like spatial extent or curvature. The possibility and limitations of measuring spatial extent by interpreting reconstructed multipole coefficients was tested under realistic noise conditions and for model misspecifications; for this analysis the primary cortical response ("N20") to electric median nerve stimulation was modeled by a one dimensional source distribution. The forward calculation was done analytically up to octapolar order for a spherical volume conductor. The multipole expansion is shown to estimate the lateral source extent with negligible bias; this estimate is to first-order stable against additional source features, like gyral curvature or spatial extent in a second direction (gyral depth, neuronal length). For a dipole moment of 20 nAm a lateral extent of 2 cm can be detected for a realistic noise level with large but experimentally still reasonable effort. Approximating a realistic head model by a sphere results in errors larger than the extent to be estimated; accordingly, studies on human cortical evoked responses will require multipole fitting in realistic head models. PMID- 11059170 TI - Ultrasound applicators with internal water-cooling for high-powered interstitial thermal therapy. AB - Internal water-cooling of direct-coupled ultrasound (US) applicators for interstitial thermal therapy (hyperthermia and coagulative thermal therapy) was investigated. Implantable applicators were constructed using tubular US sources (360 angular acoustic emittance, approximately 7 MHz) of 10 mm length and 1.5, 1.8, 2.2, and 2.5 mm outer diameter (OD). Directional applicators were also constructed using 2.2 mm OD tubes sectored to provide active acoustic sectors of 90 degrees and 200 degrees. A water-cooling mechanism was integrated within the inner lumen of the applicator to remove heat from the inner transducer surface. High levels of convective heat transfer (2100-3800 W/m2K) were measured for practical water flow rates of 20-80 mL/min. Comparative acoustic measurements demonstrated that internal water-cooling did not significantly degrade the acoustic intensity or beam distribution of the US transducers. Water-cooling allowed substantially higher levels of applied electrical power (> 45 W) than previous designs (with air-cooling or no cooling), without detriment to the applicators. High-temperature heating trials performed with these applicators in vivo (porcine liver and thigh muscle) and in vitro (bovine liver) showed improved thermal penetration and coagulation. Radial depth of coagulation from the applicator surface ranged from 12 to 20 mm for 1-5 min of sonication with 28-W applied power. Higher powers (41 W) demonstrated increased coagulation depths (approximately 9 mm) at shorter times (15 s). Thermal lesion dimensions (angular and axial expanse) produced with directional applicators were controlled and directed, and corresponded to the active zone of the transducer. These characteristic lesion shapes were also generally unchanged with different sonication times and power, and were found to be consistent with previous coagulation studies using air-cooled applicators. The implementation of water cooling is a significant advance for the application of ultrasound interstitial thermal therapy (USITT), providing greater treatment volumes, shorter treatment times, and the potential for treatment of highly perfused tissue with shaped lesions. PMID- 11059171 TI - Precision grip force dynamics: a system identification approach. AB - A linear model of the dynamics of the human precision grip is presented. The transfer function is identified as representing the peripheral motor subsystem, from the motoneuron pool to the final production of a grip force between the tip of the index finger and the thumb. The transfer function captures the limiting isometric muscle dynamics that, e.g., cortical motor areas have to act through. When identifying the transfer function we introduce a novel technique, common subsystem identification. This characterizes a specific subsystem in a complex biomechanical system. This technique requires data from two functionally different experiments that both involve the subsystem of interest. Two transfer functions, one for each experiment, are then estimated using a linear black box technique. The common mathematical factors, represented by poles and zeros, are used to form a new transfer function. It is concluded that this transfer function represents the common biological subsystem involved in both experiments. Here, we use one active and one reactive isometric grip force experiment to capture the subsystem of interest, i.e., the motoneuron pool, motor units, muscles, tendons and fingertip tissue. The characteristics of the dynamics are in agreement with previously published experiments on human neuro-muscular systems. The model, H(s) = 280/(s2 + 22s + 280), is well suited for the representation of a force producing end-effector in simulations including a control system with sensory feedback. PMID- 11059172 TI - Cryosurgical monitoring using bioimpedance measurements--a feasibility study for electrical impedance tomography. AB - The effectiveness of cryosurgery in treating tumors is highly dependent on knowledge of freezing extent, and therefore relies heavily on real-time imaging techniques for monitoring. Electrical impedance tomography (EIT), which utilizes tissue impedance variation to construct an image, is very well suited to cryosurgery since frozen tissue impedance is much higher than that of unfrozen tissue. In this study, we explore cryosurgical monitoring as a previously uninvestigated application for EIT. The feasibility of bio-impedance measurements to detect ice front propagation is demonstrated by freezing planar tissue samples one-dimensionally while measuring impedance along a linear array. The experimental results compare favorably to a simple finite element model designed to provide an electrical field visualization tool. PMID- 11059173 TI - Three-dimensional surface reconstruction and fluorescent visualization of cardiac activation. AB - Optical imaging of transmembrane potentials in cardiac tissue is a rapidly growing technique in cardiac electrophysiology. Traditional studies typically use a monocular imaging setup, thus limiting investigation to a restricted region of tissue. However, studies of large-scale wavefront dynamics, especially those during fibrillation and defibrillation, would benefit from visualization of the entire epicardial surface. To solve this problem, a panoramic cardiac visualization algorithm was developed which performs the two tasks of reconstruction of the surface geometry of the heart, and representation of the panoramic fluorescence information as a texture mapping onto the geometry that was previously created. This system permits measurement of epicardial electrodynamics over a geometrically realistic representation of the actual heart being studied. To verify the accuracy of the algorithm, the procedure was applied to synthetic images of a patterned ball; further verification was provided by application of the algorithm to a model heart placed in the experimental setup. Both sets of images produced mean registration image errors on the order of 2 pixels, corresponding to roughly 3 mm on the geometry. We demonstrate the algorithm by visualizing epicardial wavefronts on an isolated, perfused rabbit heart. PMID- 11059174 TI - Three-dimensional geometric modeling of the cochlea using helico-spiral approximation. AB - In this paper, the three-dimensional geometry of the human cochlea is modeled by the helico-spiral seashell model. The 3-D helico-spiral model, the generalized representation of the Archimedian spiral model, provides a framework for measuring cochlear features based on consistent estimation of model parameters. Nonlinear least square minimization based algorithms are developed for the identification of rotation, center and intrinsic parameters of the helico-spiral representation. Two algorithms are designed for the rotation axis aligned to the modiolar axis: one is more susceptible in the presence of noise, while the other allows applicability to two-dimensional data sets. The estimated center and intrinsic parameters allow the calculation of length, height and angular positions needed for frequency mapping of multichannel cochlear implant electrodes. Model performance is evaluated with numerically synthesized curves with different levels of added random noise, histologic data and real human cochlear spiral computed tomography data. PMID- 11059175 TI - In vivo measurement of tumor conductiveness with the magnetic bioimpedance method. AB - A noninvasive electromagnetic method has been developed that can effectively measure the in-vivo conductivity difference between rat tumor lines having a low and high metastatic potential. These tumor lines are used in the study of human prostate tumor. PMID- 11059177 TI - Gastroenterology on the Internet--IV. PMID- 11059176 TI - Neural spike sorting under nearly 0-dB signal-to-noise ratio using nonlinear energy operator and artificial neural-network classifier. AB - We report a result on neural spike sorting under conditions where the signal-to noise ratio is very low. The use of nonlinear energy operator enables the detection of an action potential, even when the SNR is so poor that a typical amplitude thresholding method cannot be applied. The superior detection ability facilitates the collection of a training set under lower SNR than that of the methods which employ simple amplitude thresholding. Thus, the statistical characteristics of the input vectors can be better represented in the neural network classifier. The trained neural-network classifiers yield the correct classification ratio higher than 90% when the SNR is as low as 1.2 (0.8 dB) when applied to data obtained from extracellular recording from Aplysia abdominal ganglia using a semiconductor microelectrode array. PMID- 11059178 TI - Red hot chilli pepper: irritating the irritable colon. PMID- 11059179 TI - Ultrasonographic assessment of gall bladder kinetics in the elderly. AB - BACKGROUND: The incidence of gallstones increases with age but the factors that influence gallstone formation in the elderly are poorly understood. Proposed factors include changes in bile composition and hypomotility of the gall bladder. Studies on gall bladder motility in the elderly have provided conflicting results, and none has been reported from India. AIM: To determine gall bladder contractility in healthy elderly subjects and compare it with that in young healthy volunteers. METHODS: Thirty healthy elderly (above the age of 60 years) and 30 young volunteers with no abdominal complaints were studied. Using real time ultrasonography and the ellipsoid method, gall bladder volume was measured after overnight fast and at 10, 20, 30, 40, 50 and 60 minutes after a standard fatty meal. Residual volume, delta volume and ejection fraction were calculated. RESULTS: Mean fasting gall bladder volume in elderly subjects was higher than that in young subjects (13.5 [5.8] mL vs 10.9 [3.6] mL; p < 0.05). However, there was no difference in the 60-min postprandial residual gall bladder volumes in the two groups. Change in gall bladder volume and ejection fraction were also similar in the two groups. CONCLUSION: There was no difference in gall bladder emptying between elderly and young subjects though the fasting gall bladder volume was higher in the elderly. PMID- 11059180 TI - Effect of cold pressor test and a high-chilli diet on rectosigmoid motility in irritable bowel syndrome. AB - AIM: Visceral hypersensitivity characterizes the irritable bowel syndrome (IBS). We evaluated the effect of a cold pressor test (CPT)--hand immersion in ice water for 1 minute, which evokes a sympathetic response--on rectosigmoid motility in patients with IBS and normal volunteers. Since many Indian patients with IBS complain of worsening of symptoms following a spicy meal, we also evaluated whether a high-chilli diet affects symptoms or changes rectosigmoid motility. METHODS: Fifteen men with IBS and 13 normal volunteers (all men) were studied. Baseline rectosigmoid manometry was done for 2 h, i.e., 1 h pre- and 1 h post CPT. The subjects were then kept in hospital on a regular diet (approximately 5 g chilli daily) for 3 days, following which symptoms were reassessed and manometry repeated for 1 h. This was followed by a high-chilli diet (approximately 15 g/day) for 3 days, after which symptoms were reassessed and manometry repeated for 1 h. RESULTS: There was no difference in the baseline study in the pre-CPT period between patients and control subjects. CPT did not change rectosigmoid motility in either group. IBS patients had varied effect on symptoms but no change in rectosigmoid motility after the high-chilli diet. In the normal volunteers, there was increased activity in the low rectum after the high-chilli diet. CONCLUSIONS: Cold pressor test does not affect rectosigmoid motility in patients with IBS or normal subjects. A high-chilli diet has varied effect on symptoms in patients with IBS and does not affect rectosigmoid motility. PMID- 11059181 TI - Use of filter paper disks for hepatitis A surveillance. AB - BACKGROUND: Venous blood collection is a cumbersome and uncomfortable procedure during hepatitis A surveillance. Collection of capillary blood by finger prick is an alternative method. AIM: To evaluate the reactivity of capillary blood/anti hepatitis A virus (HAV) IgG stored on filter paper disks for detection of anti HAV antibody. METHODS: Venous blood specimens were collected from healthy individuals. Simultaneous capillary blood specimens obtained by finger prick were stored on filter paper disks. A reference standard of anti-HAV IgG in known concentrations was spotted on filter paper disks. The reactivities of anti-HAV IgG and capillary blood specimens eluted from filter paper disks were tested by blocking ELISA for detection of anti-HAV antibody. The results were evaluated by comparing optical density (OD) and neutralization values with those obtained for WHO anti-HAV IgG stored in liquid phase and homologous venous blood specimens, respectively. RESULTS: Among both venous and capillary-blood specimens stored for 10 days, percent neutralization shown by the same 46 specimens was > 50 and that of the same 3 specimens was < 50, indicating anti-HAV positivity and negativity, respectively. There was significant correlation between the OD values displayed by anti-HAV IgG from liquid phase and that eluted from filter paper disk (p < 0.01). Sixteen serum specimens stored for a period of 2 months showed results similar to those of the corresponding filter paper disk elutes. CONCLUSION: Use of filter paper disks could be a suitable choice for pre- and post-immunization collection of blood specimens during hepatitis A surveillance. PMID- 11059182 TI - Ten-year serological follow up of hepatitis B vaccine recipients. AB - AIM: To determine long-term persistence of antibodies to hepatitis B surface antigen (anti-HBs) after vaccination against hepatitis B. METHODS: Thirty-four laboratory workers received hepatitis B vaccine in 1989 in a 0-1-6 month vaccination schedule. Group A (n = 16) received a booster at 3 years after vaccination whereas Group B (n = 18) did not. Anti-HBs was quantitated at 1 month and 1, 2, 3, 5, 6 and 8 years post-vaccination. RESULTS: At eight-year follow up, 10 of 15 subjects in Group A and 3 of 16 in Group B had protective levels of anti HBs; in addition, two and four subjects, respectively, had detectable anti-HBs though below protective levels. At ten years, 9/15 and 3/16 were anti-HBs positive in Groups A and B, respectively. One subject in each group had rise in anti-HBs titer at 6-year follow up but both of them tested negative for IgG antibodies to hepatitis B core antigen (anti-HBc). A booster dose at 10 years to anti-HBs negative subjects led to an anamestic response in 3/4 and 8/10 persons in Groups A and B, respectively. CONCLUSION: Immunological memory after vaccination against hepatitis B is maintained for at least 10 years. PMID- 11059183 TI - Acquisition of Helicobacter pylori infection and reinfection after its eradication are uncommon in Indian adults. AB - BACKGROUND: Eradication of Helicobacter pylori infection is known to decrease the recurrence rate of peptic ulcer disease. Data from India on the acquisition rate of H. pylori infection and reinfection after eradication are scant. AIM: To study the rates of acquisition of H. pylori infection and of reinfection after eradication in Indian adult patients. METHODS: We evaluated 116 consecutive patients with dyspepsia undergoing endoscopy. Sixty-four of them were H. pylori positive on gastric antral biopsy (rapid urease test and histology). Patients diagnosed to have H. pylori infection were treated with a four-drug regimen (omeprazole, bismuth subcitrate, tetracycline, furazolidine) for 2 weeks; those failing H. pylori eradication were treated with a second regimen (lansoprazole, amoxycillin, secnidazole) for one week. Patients who were H. pylori-negative to begin with and those who had successful H. pylori eradication were followed up clinically and endoscopically every 3 months for a median of one year. RESULTS: Ninety-six patients (50 H. pylori-positive) were available for study; the other 20 were lost to follow up after the first endoscopy. Fifty of the 96 (52%) were H. pylori-positive; four of these 50 patients did not follow up after first treatment. The eradication rate with the four-drug regimen was 89.1% (41/46). Four of the 5 non-responders eradicated H. pylori with the second regimen. At the end of median one year follow-up (range 9-15 months), one of the 45 patients (2.4%) who eradicated the organism developed reinfection; none of the 46 patients who were initially H. pylori-negative acquired new infection. CONCLUSIONS: The risk of reinfection after eradication is low in Indian subjects at the end of one year. The rate of acquisition of new infection is also low in the adult population. PMID- 11059184 TI - Endoscopic retrograde cholangiopancreatography in infants and children. AB - BACKGROUND: Although many reports describe the use of diagnostic endoscopic retrograde cholangiopancreatography (ERCP) in children, few mention therapeutic application of this technique in pediatric patients with pancreatic or biliary disease. METHODS: We report our 4-year experience of 80 ERCPs performed in 59 children for obstructive jaundice or cholestasis with dilatation of the biliary tree (32 children), biliary atresia (11), recurrent pancreatitis (8), and blunt trauma to the abdomen (8). RESULTS: The patients' ages ranged from 5 weeks to 18 years. The appropriate duct was cannulated in 94% of cases. Common bile duct sphincterotomy was performed in 35 patients and pancreatic duct sphincterotomy in one. Multiple procedures were done in 16 patients where biliary stents were inserted; in one patient with chronic pancreatitis and pancreaticolithiasis, pancreatic stent was inserted. Four patients developed mild pancreatitis, one had moderate pancreatitis and one had leak of contrast, which was treated by administration of clear fluids orally for one day. One patient with benign stenosis of the hepatic duct developed cholangitis after migration of the stent into the bowel lumen. CONCLUSION: Diagnostic and therapeutic ERCP can be done as safely and effectively in pediatric patients as in adults. PMID- 11059185 TI - Self-adhesive drape (Opsite) for management of leaking abdominal wounds. AB - BACKGROUND: Leaking abdominal wounds (LAW) are associated with high patient morbidity. OBJECTIVES: To evaluate the efficacy of a self-adhesive drape (Opsite) with suction drains for the management of LAW. METHODS: Twenty patients with LAW (14 intestinal fistula, 4 biliary fistula, 2 ascites leak) were subjected to the use of a self-adhesive drape with a Romovac suction drain. Conventional wound management was used for the first 5 days, followed by the application of Opsite drape. The parameters evaluated were quantity of the effluent, skin integrity, ease of application, patient comfort and cost effectiveness. A discomfort score (based on four parameters: mobility, skin excoriation, wetness and unpleasant odor) was recorded on day 1 (pre conventional), day 5 (post conventional-pre Opsite), and day 5 after Opsite application. Opsite drape was changed whenever required. RESULTS: The discomfort score was not altered with conventional therapy but was lower following Opsite application: mobility (0 vs 2), skin excoriation (0 vs 2), wetness (0.5 vs 2) and odor (0 vs 1). Opsite drape allowed accurate measurement of the effluent in all patients. The drape required change after a median of 14 days (range 10 to 18). CONCLUSIONS: Opsite drape is easy to apply on LAW, is effective in containing the effluent, and is associated with low patient morbidity. PMID- 11059186 TI - Neuroendocrine tumors of the pancreas. AB - Neuroendocrine tumors of the pancreas (NETP) are rare. We report our surgical experience of 11 patients with NETP. These included 5 patients with benign insulinomas. Raised serum insulin and C-peptide levels with hypoglycemia were always diagnostic. Ultrasonography, CT, visceral angiography, arterial stimulation and venous sampling, and intraoperative ultrasound localized the tumor in 0/5, 1/5, 3/4, 2/2, 3/3 cases, respectively. The 6 other malignant NETP (one gastrinoma, 2 carcinoids, 3 non-functioning) were managed by pancreatic resection (Whipple's operation = 3, distal pancreatectomy with total gastrectomy = 1, total pancreatectomy = 1, distal pancreatectomy with left nephrectomy and proximal gastrectomy = 1). Two patients died postoperatively. We had 5 major and 2 minor postoperative complications, with 2 deaths. During follow up from 1 to 7 years, one patient with malignant carcinoid tumor died at two and half years, of local recurrence. The other 8 patients are disease-free with good quality of life. PMID- 11059187 TI - Isolated splenic lymphoma: an elusive preoperative diagnosis. AB - Four patients underwent splenectomy for various clinical and radiological diagnoses and were found to have primary splenic lymphoma at surgery and histology. The diagnosis was classical Hodgkin's lymphoma, mixed cellularity type (one case); marginal zone B-cell non-Hodgkin's lymphoma (one case); and large B cell type non-Hodgkin's lymphoma (two cases). The first two patients had multiple nodules in the spleen measuring 0.1-0.5 cm while large cell lymphomas had large nodules (largest measuring 11 cm x 7 cm x 4 cm). The diagnoses were confirmed by immunohistochemical analysis. Mean follow up of these patients was 11 months; all patients received chemotherapy. One patient died, of causes not related to the disease process. PMID- 11059188 TI - Leiomyosarcoma of pancreas: imaging features. AB - Leiomysosarcoma of the pancreas is rare. Radiological findings of four patients with this condition were reviewed. CT scan appearances were those of a predominantly homogenous, large, solid, enhancing mass with few areas of necrosis. A cystic appearance with thickened and nodular enhancing walls was seen in one case. A constellation of imaging findings including large size at presentation, greater vascular enhancement and absence of biliary dilatation should suggest leiomyosarcoma. PMID- 11059189 TI - Occult primary adenocarcinoma causing pseudomyxoma peritonei with pleural and hepatic metastases. AB - Pseudomyxoma peritonei is characterized by diffuse collections of gelatinous flux resulting from implantation of malignant tumors or irritation from ruptured benign cysts. We report a patient with pseudomyxoma peritonei caused by an occult primary adenocarcinoma, who had both pleural and hepatic metastases. PMID- 11059190 TI - Volvulus and gangrene in intra-abdominal colon after colonoplasty for esophageal stricture. AB - Ten years after colonoplasty using the left colon for impassable corrosive stricture of the esophagus, a 27-year-old woman developed volvulus and gangrene of the remaining intra-abdominal colon. It was resected and colostomy was done. In the second stage, after 3 months, the ascending colon and cecum were mobilized and anastomosed to the rectal stump. Eight years later, the patient is asymptomatic. PMID- 11059191 TI - Bleeding scrotal varices as presentation of Budd-Chiari syndrome. AB - Budd-Chiari syndrome presents with ascites, edema and bleeding from esophageal varices. Presentation as bleeding scrotal varices is rare. We report a patient with Budd-Chiari syndrome who presented with recurrent bleeding from scrotal varices for 20 years. PMID- 11059192 TI - Paterson-Kelly syndrome and celiac disease--a rare combination. AB - Paterson-Kelly syndrome is characterized by an association of iron deficiency with dysphagia. We describe a patient with this syndrome who was later diagnosed to have celiac disease. PMID- 11059193 TI - Neural hypertrophy in carcinoma stomach. AB - Neural hypertrophy with hyperplastic Schwann cells in the wall of the stomach along with enterochromaffin cell hyperplasia was incidentally observed at histology in the gastrectomy specimen of a 43-year-old man with carcinoma stomach who had presented with upper abdominal pain of one year duration. The patient had no previous abdominal surgery or evidence of gastrointestinal obstructive pathology. The significance of this neural hypertrophy is not known. PMID- 11059194 TI - Vesical varix in cirrhosis of liver. AB - Vesical varices in portal hypertension are rare. We report a patient with portal hypertension who developed recurrent painless hematuria. Cystoscopy was normal. Doppler ultrasound and MR angiography showed a dilated paraumbilical vein within the falciform ligament coursing down to the urinary bladder wall and draining into the right internal iliac vein. He underwent liver transplantation for decompensated chronic liver disease. He is in good health and has not had further episodes of hematuria. PMID- 11059195 TI - VIPoma of pancreas in a child. AB - An eleven-year-old girl had massive watery diarrhea. She was found to have pancreatic VIPoma. It responded favorably to surgical resection of the tumor. There was no tumor recurrence at 18 months of follow-up. PMID- 11059196 TI - Acute pancreatitis in a child with idiopathic ulcerative colitis on long-term 5 aminosalicylic acid therapy. AB - Acute pancreatitis is a rare but known complication of inflammatory bowel disease in adults. In children, only a few cases with this complication have been reported. We describe a 10-year-old boy with ulcerative colitis who developed acute pancreatitis while on long-term treatment with 5-aminosalicylic acid. PMID- 11059197 TI - Recombinant interferon-alfa therapy in children with chronic hepatitis B and cured cancer. PMID- 11059198 TI - Hepatitis B virus: healthy carrier or chronic infection? PMID- 11059199 TI - Hepatitis B virus: healthy carrier or chronic infection? PMID- 11059200 TI - Gallstone ileus presenting as malignant jejunal obstruction. PMID- 11059201 TI - Professionals' use of different mentor sources at various career stages: implications for career success. AB - The authors investigated the various sources of mentors used by professionals, how these sources influenced both objective and subjective career success, and whether the participants used different sources of mentors at different stages of their careers. According to data from 430 faculty members at 2 U.S. research institutions, assistant professors with mentors in their professions, associate professors with mentors outside the work place, and professors with mentors within their organizations had the highest levels of objective career success. Assistant professors with multiple sources of mentors yielded significantly higher levels of both objective and subjective career success than did those with single sources or no mentor. If one links professorial rank to career stage, the results suggest that the participants used different sources of mentors at different stages of their careers. PMID- 11059202 TI - Abstinence and well-being among members of Alcoholics Anonymous: personal experience and social perceptions. AB - The authors examined the subjective experience of well-being (WB) among abstinent Alcoholics Anonymous (AA) members and social perceptions of an abstinent alcoholic's WB among 3 nonalcoholic French-Canadian samples: male police officers, Catholic nuns, and university women. The short-term abstinent AA members, along with the university women, reported the lowest self-ratings of WB, whereas the Catholic nuns reported the highest. However, among the abstinent AA members, the level of WB was positively related to the length of abstention. The 3 nonalcoholic groups evaluated an abstinent AA member more positively than a nonabstinent alcoholic. These evaluations of an abstinent AA member converged with the AA members' self-evaluations on the measure of WB. PMID- 11059203 TI - The effect of children's sleeping arrangements (communal vs. familial) on fatherhood among men in an Israeli kibbutz. AB - The author examined the effect of children's sleeping arrangements (communal vs. familial) on the extent of fathers' involvement in their children's lives and their level of satisfaction from fatherhood. Questionnaires assessing those aspects of fatherhood were administered to 40 fathers living in a kibbutz. Results indicated that the fathers of children sleeping at home were more involved with their children's lives and showed higher levels of satisfaction from fatherhood. Preference for a communal sleeping arrangement for children was expressed among 7 men, all of whom were older than 50 years. These results are discussed in the context of culturally changing concepts of paternal roles in the family and socioeconomic transition in the kibbutz. PMID- 11059204 TI - Perceptions of dual identity and separate groups among secular and religious Israeli Jews. AB - The authors examined the effects of perceptions of dual identity and separate groups on tendencies to handle intergroup conflict through problem solving and contention. Among secular Israeli Jews, regression analyses revealed a significant interaction between perceptions of dual identity and perceptions of separate groups: Only under high perception of dual identity was the perception of separate groups associated with contention. Among religious Israeli Jews, problem solving and contention were unrelated to either dual identity or to perceptions of separate groups. The results are discussed in terms of the common ingroup identity model (S. L. Gaertner, M. C. Rust, J. F. Dovidio, B. A. Bachman, & P. A. Anastasio, 1994) and in the context of the conflict between religious and secular Jews in Israel. PMID- 11059205 TI - Work values among Lebanese workers. AB - On the basis of a review of the existing literature, the authors tested 4 hypotheses to determine the applicability of work values in Arab societies to employees in Lebanese organizations. Only 1 hypothesis was supported: Organizational policies that ran counter to the worker's religious values had an adverse effect on job satisfaction. There was no support for the hypotheses (a) that workers' religiosity in inversely related to positive attitudes toward women's involvement at work, (b) that employee satisfaction is related to a mechanistic organizational design, or (c) that workers with an internal locus of control experience higher job satisfaction. The Lebanese workers, thus, did not appear to share some of the attributes claimed to exist in Arab societies. PMID- 11059206 TI - Self-efficacy and job-seeking activities in unemployed ethnic youth. AB - The authors examined variables predicting the self-efficacy and the job-seeking activities of 103 unemployed ethnic youth from diverse cultural backgrounds. Whereas self-esteem and acceptance by the host culture were significant predictors of self-efficacy, the major predictors of job seeking were (a) the extent to which the ethnic youth felt accepted by members of the dominant culture and (b) the difference between their cultural backgrounds and the dominant host culture. Implications of these findings for enhancing the employment of ethnic youth are discussed. PMID- 11059207 TI - Development and validation of a scale measuring attitudes toward smoking. AB - The authors developed the Smoking Attitudes Scale (SAS) and administered it to 2 samples of U.S. students who were smokers or nonsmokers. Exploratory and confirmatory factor analysis with LISREL (K. G. Joreskog & D. Sorbom, 1989) methodology revealed that the SAS consists of 4 factors. The overall instrument possesses good internal consistency and adequate construct validity as well. PMID- 11059208 TI - Birth order and civil disobedience: a test of Sulloway's "Born to Rebel" hypothesis. AB - In Born to Rebel, F. Sulloway (1996) argued that, throughout history, later-borns have been more likely than first-borns to challenge the status quo. The authors tested Sulloway's hypothesis among a group of U.S. college students who had participated in civil disobedience as part of a labor dispute. The authors predicted that there would be a higher percentage of later-borns among those who had been arrested than among a group of their friends who had not participated in civil disobedience or among a control group of students drawn from classes at the college. The findings, in fact, revealed a significant relationship between the number of times the students had been arrested and birth order. PMID- 11059209 TI - Confirmatory factor analyses of the full and short versions of the Marlowe-Crowne Social Desirability Scale. AB - Over the years, researchers have developed various short versions of the Marlowe Crowne Social Desirability Scale (D. P. Crowne & D. Marlowe, 1960). The authors used confirmatory factor analyses (J. L. Arbuckle, 1997) as well as item and scale analyses to evaluate the adequacy of the full version and various short versions. Overall, the results from 232 Canadian undergraduates showed (a) that all the short versions in the present study are a significant improvement in fit over the 33-item full scale and (b) that W. M. Reynolds's (1982) Forms A and B are the best fitting short versions. No gender differences were found for the full scale or any of the short versions. The results show that the full scale could be improved psychometrically and that the psychometrically sound short versions should be available because they require less administration time than the full scale. PMID- 11059210 TI - The commodified self in consumer culture: a cross-cultural perspective. AB - The author examined the premise that the pervasive consumer ideology in today's society has implications for the conceptions of self and others. She used structured interviews to assess conceptions of self and other and G. P. Moschis's (1978) questionnaire to measure consumer orientation among 76 U.S. and 62 Finnish college students. The principal hypotheses were (a) that the U.S. students would be more commodified than the Finnish students and (b) that the participants who were high in consumer orientation would be more likely than those lower in consumer orientation to see themselves and others in material terms and also to consider personal characteristics to have market values. The results supported both hypotheses. PMID- 11059211 TI - Understandings and experiences of cruelty: an exploratory report. AB - The authors investigated popular understandings of cruelty among 103 undergraduates who identified the cruelest acts that they had experienced vicariously and personally. The authors also examined the reasons that the cited acts were defined as cruel. Results indicated that most of the vicarious cruel acts involved intense aggression or sexual imposition, whereas personally experienced cruelty was milder, frequently consisting of teasing or gossip. Offense, victim, and perpetrator characteristics were all cited as reasons that acts were considered cruel. The authors also investigated gender differences in reported acts and reasons. Future researchers should address the discrepancies between vicarious and personally experienced cruelty. Findings with regard to personal acts also call for links to the literature on callousness and victimization. PMID- 11059212 TI - Italian, Bulgarian, and U.S. children's perceptions of gender-appropriateness of occupations. PMID- 11059213 TI - The relationship between parental control and scholastic achievement of children from single- and two-parent families. PMID- 11059214 TI - Depressive symptoms among U.S. and Indian college students: the effects of gender and gender role. PMID- 11059215 TI - Recent advances in hepatitis C. AB - Nearly 2% of Americans are infected with the hepatitis C virus, resulting in 8,000-10,000 deaths each year. The virus can be eradicated with interferon plus ribavirin in about 40% of patients treated for one year. Patients with a history of intravenous drug abuse or transfusion before July 1992 should be screened for hepatitis C, as should members of other high-risk groups. Infected patients should be counseled and if possible offered treatment. This will prevent unnecessary deaths from liver failure and hepatocarcinoma. PMID- 11059216 TI - The burden of cancer in Kentucky. The 1998 Kentucky cancer incidence report. AB - The purpose of collecting this important disease burden information is just for reports like this. The real value lies in use of data for cancer control research, program planning, resource allocation, program design, and evaluation. Data in the annual KCR report and on the website should be a valuable resource for health agencies, clinicians, policymakers, voluntary organizations, etc in their assessment of health problems in their area and as critical decisions are made about how to utilize limited intervention resources. The Kentucky Cancer Registry will continue to work very closely with the Kentucky Cancer Program, Kentucky Medical Association, and programs and research efforts at the University of Kentucky Cancer Control Program (Mid South Cancer Information Service, Appalachia Cancer Network, Kentucky Prevention Research Center) and the Brown Cancer Center at the University of Louisville to make these data available to partners throughout the Commonwealth to help guide their assessment and planning processes. If you have questions about this data you are encouraged to contact Regional Coordinators for the Kentucky Cancer Program in your Area Development District. The contact information is shown in Appendix I. PMID- 11059217 TI - Electronic distractions ... a medical concern? PMID- 11059218 TI - SAVE--school violence II. PMID- 11059219 TI - Anticancer drugs induce apoptosis in mouse hair follicles. AB - Apoptosis represents an important mode of cell death induced by chemotherapeutic drugs. It is plausible that apoptosis may also play an important role in alopecia caused by anticancer drugs. C57 BL/6 mice were induced to enter into anagen by hair shaft depilation and a single intraperitoneal injection of cyclophosphamide (150 mg/kg) was given. Skin specimens were taken from the backs of mice at day 9, 10, 11, 12 and 13 postdepilation (p.d.). Specimens were stained with TUNEL method, and DNA was isolated from the plucked hair follicles and was electrophoresed. Many TUNEL-positive cells were observed in hair bulbs in the cyclophosphamide group at days 10-13. At day 13 p.d., most follicles were damaged and extensive hair loss was observed. Fragmented DNA, but not so distinct, were observed in a DNA ladder pattern corresponding to the in situ results. This study suggests that anticancer drugs can induce apoptosis-related damage to the hair follicles in mice. Control mice when treated with saline also showed apoptosis when they spontaneously entered into catagen, but they never showed hair loss. Consequently, hair loss induced by anticancer drugs may result from a wave of induced apoptotic death. PMID- 11059220 TI - Immunohistochemical study on the expression of cyclin D1 and E in gastric cancer. AB - Cyclin D1 and E have been found to be deregulated and overexpressed in various types of cancers. In order to study the cell cycle regulatory mechanisms in gastric cancer, we have analyzed the protein expression of cyclin D1 and cyclin E in 76 tumor specimens from patients with primary gastric cancer, using immunohistochemistry. Overexpression of cyclin D1 was observed in 38 cases (50.0%). Overexpression of cyclin E was observed in 40 cases (52.6%). There was no significant difference between the expression of cyclin D1 and any clinicopathological factor. Cyclin E overexpression was correlated with a high incidence of lymph node metastasis, a low incidence of T1, and with Stage I. There was no significant difference in survival curves between cyclin D1 (+) and cyclin D1 (-). The survival curves of cyclin E (-) were significantly higher than those of cyclin E (+). These results suggested that in gastric carcinoma, cyclin E overexpression was useful as a prognostic indicator, but cyclin D1 was not. PMID- 11059221 TI - In vitro susceptibilities to 23 antimicrobial agents of Haemophilus influenzae from pediatric patients in Japan. AB - One hundred eighty-six clinical isolates of Haemophilus influenzae (H. influenzae) collected from January 1996 through December 1997 from 182 pediatric patients and 16 isolates from blood or cerebrospinal fluid (CSF) of 13 patients with bacteremia and purulent meningitis collected during the last ten years were examined for in vitro susceptibilities to 23 antibiotic agents, including 3 penicillins, 9 cephalosporins, 4 carbapenems and others, as well as for their encapsulated types and beta-lactamase production. Ceftriaxone (a third generation cephalosporin) had the highest activity against the strains in this study (minimal inhibitory concentration, MIC90 of 0.025 microgram/ml) and cefditoren (a new oral cephalosporin) was the most active oral antimicrobial agent (MIC90 of 0.05 microgram/ml). Meropenem had a much higher activity against H. influenzae (MIC90 of 0.2 microgram/ml) than the other carbapenems (imipenem, MIC90 of 1.56 micrograms/ml, panipenem, MIC90 of 1.56 micrograms/ml, and biapenem, MIC90 of 3.13 micrograms/ml). Regarding the serotyping of the encapsulated strains, 172 strains (85.1%) were nontypeable and 30 (14.9%) were serotyped (24 strains of type b, 4 strains of type e, one each of type a and c). Fifteen of the strains isolated from blood and CSF were type b and one was nontypeable. Sixteen of 202 strains (7.9%) produced beta-lactamase and all of them produced both penicillinase and cephalosporinase. The production of beta-lactamase in this study was lower than that reported in previous studies [1-3]. In this study, some strains were found against which the MICs of carbapenems were very high (highest MIC of imipenem was 12.5 micrograms/ml, of panipenem was 6.25 micrograms/ml and of biapenem was 25 micrograms/ml). Therefore, we assayed the binding affinities of imipenem for each of penicillin-binding proteins (PBPs) about one of these resistant strains. In resistant strains, inhibitory concentrations (IC50) of imipenem for PBP4 and 5 were much higher than those in susceptible strains. Thus, the results demonstrate the decrease of the affinity of imipenem for PBP4 and 5. It seems, therefore, that the major factor in the resistance to imipenem of H. influenzae was the low affinity of PBP4 and 5 for the drug. PMID- 11059222 TI - Biomechanical evaluation of foot pressure and loading force during gait in rheumatoid arthritic patients with and without foot orthosis. AB - Foot orthoses are commonly used in patients with rheumatoid arthritis (RA) to support the foot and relieve pain, however little is known about the biomechanical effects of in-shoe foot orthoses in reducing or redistributing high pressures and loading forces. The purpose of this study was to compare the foot pressures and loading forces during gait in rheumatoid arthritic patients and healthy subjects, and evaluate the biomechanical effects of the foot orthoses in the RA patients. Twelve female RA patients with foot pain in walking, all Steinbrocker class II, and 8 healthy women without foot pain were matched for age. Foot pressures and loading forces with and without orthoses were measured using the F-Scan program. The pressure distributions and loading forces were standardized by the body weight and compared, and the effects of the foot orthoses were evaluated. The foot orthoses of RA patients provided higher pressure reduction than those of the control group (3.00 +/- 0.38 g/cm2/BW and 3.29 +/- 0.29 g/cm2/BW respectively, p < 0.001). Similar redistribution of plantar pressures and loading forces were found between two groups but the RA patients had a greater change at the stance phase of gait (p < 0.0001). The foot orthosis produces greater pressure and loading force relief and redistribution in RA patients than in normal subjects. PMID- 11059223 TI - Efficacy of intermittent etidronate therapy for corticosteroid-induced osteoporosis in patients with diffuse connective tissue disease. AB - We conducted a one-year comparative study of 25 patients with corticosteroid induced osteoporosis associated with diffuse connective tissue disease. The patients were randomly divided into 2 groups: group A (9 patients), monotherapy with active vitamin D3 (V.D3); and group B (16 patients), combination therapy with V.D3 and etidronate. Four markers were employed: as an bonegenic marker, serum alkaline phosphatase (ALP); as a bone resorption marker, urinary deoxypyridinoline (DPD); as a bone salt minerals assay level, young adult mean (YAM); and bonefracture ratio. Results showed that: ALP decreased in both groups with no significant difference between groups; DPD increased significantly from baseline (p < 0.05) in group A, but it decreased significantly from baseline (p < 0.05) in group B, but again without a significant difference between groups; YAM resulted in no significant improvement in group A, but a significant improvement from baseline (p < 0.01) was shown in group B, with a significant difference between groups (p < 0.05); and a new spinal compression fracture ratio was extremely lower in group A than in group B. The findings indicated cyclical/intermittent etidronate therapy is effective in preventing corticosteroid-induced osteoporosis. PMID- 11059224 TI - Delayed CSF pseudocyst following shunt malfunction after myelomeningocele repair. AB - We report a case of delayed cerebrospinal fluid (CSF) pseudocyst following shunt malfunction in a 20-year-old patient with myelomeningocele. Magnetic resonance (MR) images and a radioimmunoassay shunt-gram detected the CSF fistula at the old scar of the myelomeningocele repaired 20 years before. Since the revision of the shunt system failed to keep the pseudocyst, the lesion was successfully directly repaired. Treatment of the delayed CSF pseudocyst following shunt malfunction thus requires a radical operation of the lesion. PMID- 11059225 TI - A case of duodenal papillary carcinoma complicated by repeated acute pancreatitis. AB - We present a patient with duodenal papillary carcinoma who repeatedly developed acute pancreatitis preoperatively. The patient was a 65-year-old male. In February 1997, the patient consulted a local hospital due to vomiting, high fever, and jaundice. With the diagnosis of obstructive jaundice, percutaneous transhepatic biliary drainage (PTBD) was performed, revealing a distal bile duct obstruction. Because duodenal papillary carcinoma was diagnosed based on endoscopic findings, the patient was admitted to Kurume University Hospital. Hypotonic duodenography (HDG) disclosed a protruding lesion with an irregular surface in the descending part of the duodenum, resulting in a diagnosis of positive duodenal invasion (du1). Because computed tomography (CT) demonstrated a protruding lesion on the medial side of the second portion of the duodenum, positive pancreatic invasion (panc2) was diagnosed. On March 18 and April 22, sudden abdominal pain, leukocytosis, and an increase in serum amylase were noted. CT revealed that the pancreas was diffusely enlarged, showing an ill-defined boundary between the pancreas and adipose tissue and fluid collection. On CT, the lesion was evaluated as Grade 3 and moderate. For treatment, pancreatic enzyme inhibitors and antibiotics were intravenously injected. Peritoneal perfusion was concomitantly performed during the second treatment. Because symptoms remitted thereafter, a pylorus preserving pancreatoduodenectomy (PpPD) was carried out. The postoperative histologic examination revealed negative pancreatic invasion. Concerning the etiology of acute pancreatitis, not pancreatic invasion, but impaction of the liberated tumor mass in the common canal was considered responsible for the repeated pancreatitis because the tumor showed a cauliflower like shape. PMID- 11059226 TI - A case of intrahepatic gallstone formation around nylon suture for hepatectomy. AB - A 69-year-old female underwent left lobectomy for hepatolithiasis in February 1994. She was admitted to the Kurume University Hospital in December 1997 because computed tomography (CT) showed calcification in the porta hepatis. Ultrasonography (US) revealed a hyperechoic area with an acoustic shadow in the right hepatic duct. Dilated intrahepatic bile ducts and a mural lucent area in the right hepatic duct were noted on endoscopic retrograde cholangiography (ERC). Although the above findings suggested a diagnosis of recurrent hepatolithiasis, percutaneous transhepatic biliary drainage (PTBD) for biopsy was performed in order to rule out cancer. Biopsy showed no evidence of malignancy. Under a cholangioscope, a tip of a nylon suture was found to be protruding into the bile duct. Although a gallstone had already slipped off, the surface of the nylon suture was covered with biliary sludge. The protruding tip of the nylon suture was considered to be the nucleus of the stone. The tip was removed under cholangioscopy. Postoperative CT confirmed the absence of calcification in the porta hepatis. There has been no recurrence of hepatolithiasis after surgery. Although the formation of gallstones around the core of nylon sutures is very rare, absorbable sutures should be used during surgery of the bile duct because nonabsorbable sutures can become the nucleus of gallstones. PMID- 11059227 TI - Bilateral pleural effusion following cervical abscess drainage: a case report. AB - Postoperative bilateral chylothorax after cervical surgery has been rarely reported, whereas unilateral chylothorax has been occasionally reported after thoracic surgery. Here, we report a rare case of bilateral pleural effusion that developed after cervical abscess drainage. On the second day after the drainage, the patient felt dyspnea, and bilateral pleural effusion was found on a chest X ray. The effusion was thought to be chyle and was successfully treated with conservative management. Additionally here, we have suggested that non-traumatic chylothorax was caused by increasing intraluminal pressure occurring inside the thoracic duct after its ligation. Careful follow up of any respiratory symptoms and of chest X-rays is recommended after cervical intervention. PMID- 11059228 TI - Two cases of cancer in the remnant stomach derived from gastritis cystica polyposa. AB - We have recently encountered two patients with early gastric cancer in the remnant stomach which resulted from gastritis cystica polyposa at the anastomosis site. The remnant stomach, which had been reconstructed with the Billroth II method, contained an elevated sessile lesion at the anastomosis site. One patient was a 73-year-old woman who had undergone gastrectomy for a gastric ulcer at 30 years earlier, cancer type I + IIa of the remnant stomach was diagnosed, and total remnant gastrectomy was performed. The other patient was a 59-year-old man who had undergone gastrectomy for a duodenal ulcer at 31 years earlier, cancer type I + IIa of the remnant stomach was diagnosed, and subtotal remnant gastrectomy was performed. Histological examination in each case showed that moderately differentiated adenocarcinoma had developed from gastritis cystica polyposa. These results suggested that this cancer has a close relationship with gastritis cystica polyposa. PMID- 11059229 TI - Two anomalous cases of the hepato-mesenteric and the gastro-splenic trunks independently arising from the abdominal aorta. AB - This report describes two arterial anomaly cases in the celiaco-mesenteric region, which were encountered in two Japanese male cadavers in the dissecting room at Kurume University School of Medicine in 1999. In these cases, the usual celiac trunks were not identified, and the hepato-mesenteric and the gastro splenic trunks were independently arising from the abdominal aorta. Moreover, in the first case, the common hepatic artery passed ventral side of the portal vein and divided into the hepatic proper and the gastroduodenal arteries. This type of arterial anomaly belongs to the Type V of Adachi's classification and the Type IV' of Morita's classification. In the second case, the common hepatic artery passed dorsal side of the portal vein and divided into the hepatic proper and the gastroduodenal arteries. This type of arterial anomaly belongs to the Type VI of Adachi's classification and the Type IV' of Morita's classification. PMID- 11059230 TI - Understanding and awareness of irradiated food in Japanese young students. PMID- 11059231 TI - [Videolaparoscopic cholecystectomy. Experience and results in 1019 cases]. AB - BACKGROUND: Laparoscopic cholecystectomy is now worldwide considered the elective therapy for biliary lithiasis. Morbidity, mortality and conversion rates reported in the literature are assessed between 1 and 6%, 0 and 0.9%, 3.6 and 7.2% respectively. Data on personal experience with 1019 attempted laparoscopic cholecystectomy are reported. METHODS: In the period between 1991 and 1997 1019 laparoscopic cholecystectomy were performed. Patients were 361 males and 658 females (ratio M:F 1:2), with an average age of 51 years (range 5-85). Indications were: 647 symptomatic cholelithiasis, 28 hydrops, 121 empyemas, 76 cholecystocholedocolithiasis and 13 alithiasic cholecystopathy. RESULTS: Conversion was necessary in 61 cases, with a conversion rate of 6%. Mean duration of surgery was 65 minutes (range 30-240) with a mean hospital stay of 2.1 days (range 1-10). No deaths occurred in our series, with a morbidity rate of 1.8% (18 cases, 7 major and 11 minor). Only 1 case of bile duct injury (0.1%) is reported. CONCLUSIONS: In consideration of low conversion rate, low early and late morbidity, absence of bile duct injury, advantages for the patient and the opportunity of evolution of this surgery, laparoscopic cholecystectomy can be considered the standard treatment for biliary lithiasis. PMID- 11059232 TI - [Hemorrhoidectomy in muco-hemorrhoidal prolapse using mechanical stapler]. AB - BACKGROUND: The treatment of hemorrhoids has changed throughout the latter years. In selected cases hemorrhoidectomy with circular stapler, in personal opinion, is the best surgical technique. The goal of the present study is to evaluate the therapeutic results of hemorrhoidectomy by means of the above technique in 150 consecutive cases. METHODS: From March 1997 to November 1999, 150 patients were surgically treated, 84 women and 66 men with a median age of 57 years. 130 patients had III degree, thus with a reducible prolapse, 10 IV degree and 10 patients with II degree hemorrhoids resistant to ambulatory banding and/or sclerotherapy. Every patient was operated with the circular stapler in Day Surgery regimen, and the patients were discharged after 24 hours. RESULTS: Eight immediate hemorrhages were seen, four of which were surgically treated; 10 patients had urinary retention which required an urinary catheter. Healing was complete in the 15-20th day. The follow-up after one month was performed on 140 patients, at 3 months and at one year, on 100 patients. No suture stenosis and no recurrences of the hemorrhoid prolapse were observed. Follow-up after 6, 12 and 24 months on 100 patients was also considered in order to evaluate the results after surgery and late complications or sequelae. Fifteen patients were reevaluated after one and two months with anorectal manometry which did not demonstrate important alterations of the continence. CONCLUSIONS: The advantages of this technique is to carry out a radical surgical procedure in a single event, by means of a rapid and mini-invasive technique, with an earlier physical recovery, with the absence of medication in the postoperative period, and an early social and working reestablishment. PMID- 11059233 TI - [Surgical treatment of hemorrhoidal disease using circular mechanical stapler. Analysis of costs]. AB - BACKGROUND: An analysis of surgical treatment costs of haemorrhoid disease with the use of a new circular stapler, is made comparing this operation with Milligan Morgan's technique. The money and management saving due to the reduction of operation time and postoperative stay compensates present costs due to stapler. METHODS: 35 uniform patients (number, age, sex, grade of haemorrhoid disease and surgical equipe) are considered and divided into two groups of study to evaluate perspectively the surgical costs. RESULTS: The cost of treatment with stapler per patient is like Milligan Morgan's treatment (1.714.681 lire versus 1.681.893), with an important management saving of postoperative days (16 hours versus 42 hours). CONCLUSIONS: Moreover there are psychologic and social advantages, not quantifiable but considerable, due to the early physical recovery and to the absence of out-patient dressing cycles peculiar of this surgical technique, with a rapid social and working reinstatement of patients (4-5 days versus 4-5 weeks of conventional intervention). PMID- 11059234 TI - [Results of preoperative staging using endosonography in rectal cancer]. AB - BACKGROUND AND AIM: The latest reports using transrectal ultrasound (TRUS) for the preoperative staging of rectal cancer show a diagnostic accuracy between 78 and 97% with regard to the local spread of disease, and between 62 and 86% for the diagnosis of lymph node metastasis. The correct choice of surgery depends on correct preoperative staging, as does the indication for any preoperative neoadjuvant treatment. The aim of this study was to evaluate the diagnostic precision of the method used by the authors since 1993 by comparing the preoperative transrectal ultrasound stage (uTMN) with the postoperative histological stage (pTNM). In addition, the study aimed to assess whether some anatomic and pathological characteristics of the neoplasm (differentiation, type of growth and presence of peritumoral inflammatory reaction) influenced the diagnostic precision of transrectal ultrasonography. METHODS: Forty-two patients with a preoperative histological diagnosis of adenocarcinoma localised in the rectal segment, extending up to 10 cm from the dentate line, undergoing radical surgical were selected from the group of patients with middle-lower rectal cancer studied preoperatively with TRUS. Preoperative TRUS was carried out in 42 cases by a single examiner. Anatomic and pathological examination of the removed portion was performed by examiners who were not familiar with the preoperative ultrasonographic diagnosis. RESULTS: In this study TRUS showed a diagnostic accuracy of 81% in the study of T and 71.4% in the study of N. In line with other studies, the most frequent diagnostic error was the overstaging of stage T2 tumours. Moreover, the presence of a peritumoral inflammatory reaction was found to be the only variable that significantly influenced the diagnostic accuracy of TRUS. CONCLUSIONS: TRUS was found to be a valid instrument for the preoperative staging of rectal cancer even in this preliminary study limited to 42 cases, in particular with regard to wall invasion. The limits of this method are linked to the presence of phenomena producing a situation of local infection (recent biopsies, radiotherapy, peritumoral inflammatory infiltrate) given that this prevented the correct visualisation of the layers of the rectal wall. As a result, this may limit its use in the re-staging of patients undergoing preoperative radiotherapy. PMID- 11059235 TI - [Sliding hiatal hernia in patients with gastroesophageal reflux: physiopathology and surgical treatment]. AB - BACKGROUND: The aims of the study were to evaluate how the sliding hiatal hernia, in patients with gastroesophageal reflux disease (GERD), acts on the lower esophageal sphincter (LES) and esophageal clearance, and how surgical therapy corrects the physiopathological parameters. METHODS: Records of 25 patients with only GERD and of 15 with GERD associated to hiatal hernia (> 3.5 cm) were reviewed. Ten subjects without symptoms and/or endoscopic and functional signs of GERD were considered as control group. The selection of the patients was done by reviewing radiographic examination, endoscopy and functional tests (esophageal manometry, pH-monitoring). RESULTS: Manometry showed a greater LES incompetence (pressure and length) and a worse peristalsis (distal amplitude) in the group with reflux and hiatal hernia against patients with reflux only. Also, patients with hiatal hernia had more acid exposure (total time pH < 4 in the distal esophagus) and a longer time of esophageal clearance, at pH-monitoring. The functional tests in 8 patients, before and after laparoscopic Toupet fundoplication with posterior closing of the crura, showed a normalised LES, esophageal clearance and acid exposure. Esophageal peristalsis did not show any statistically significance. CONCLUSIONS: The presence of hiatal hernia, in patients with GERD, causes worse LES, peristalsis and clearance with a greater acid exposure of the esophagus. Fundoplication, by reconstructing the sphincter diaphragm unit, normalises the preoperative physiopathology situation but without an effective peristalsis improvement. PMID- 11059236 TI - [Diagnosis of severity as a basic parameter in the treatment of acute biliary pancreatitis]. AB - BACKGROUND: Acute biliary pancreatitis (ABP) still retains high morbidity (15 50%) and mortality (20-35%). Therefore it appears to be crucial to clearly assess the aetiological factors (50% of idiopathic are in fact biliary pancreatitis) and to establish their severity in order to plan the appropriate treatment. METHODS: 58 ABP patients were diagnosed by ultrasound (77.5%) or by laboratory findings (22.4%). Following Ranson and APACHE II scoring 17 cases (29.3%) were classified as severe, 41 (70.6%) as mild. All patients with severe ABP, had emergency ERCP + ES (within 24-48 hrs) followed by LC (< or = 10 days). Patients with mild ABP had LC within 10 days; in these cases IOC was always done. RESULTS: In severe cases operative endoscopy cured pancreatic inflammation in 12 cases. Subsequent LC never showed serious morbidity, apart from subcutaneous emphysema in one case. In 5 cases laparotomy was required since pancreatic necrosis was present, with 60% mortality. In patients with mild pancreatitis LC was successfully performed in all cases, with 7.3% morbidity. IOC showed choledochal stones in 31.7% of cases, while in severe cases stones in the biliary tree were shown in 88.2% of cases. CONCLUSIONS: In conclusion ABP treatment is always surgical, and almost always with minimally-invasive procedures in severe cases (ERCP + ES with LC < or = 10 days) if surgery is performed within 24-48 hrs as well as in mild cases (LC + IOC) when surgery is done within 10 days. PMID- 11059237 TI - [Intestinal fistulas in Crohn disease]. AB - BACKGROUND: Only a part of patients suffering from Crohn's disease has enteric fistulae and a different behaviour of Crohn's disease with fistulae is reported in the literature. Aim of this paper is to evaluate if enteric fistulae are a factor conditioning mortality, morbidity and overall postoperative course, in patients with Crohn's disease. METHODS: Data on the postoperative course of 126 laparotomies for Crohn's disease, performed between November 1993 and July 1998, have been prospectively examined. Moreover, the presence of enteric fistula has been evaluated during surgery. RESULTS: Out of 126 interventions, in 58 (46%) enteric fistulae were present. Mortality (5.2% vs 0), morbidity (14.5% vs 7.3%), necessity for a temporary ostomy (20.4% vs 3.5%) were greater in those patients with fistula, as compared as those without fistula. CONCLUSIONS: In conclusion, it is suggested that Crohn's disease with fistulae is a different type of disease, with higher mortality and morbidity rates. PMID- 11059238 TI - [Spigelian hernia: an up-to-date]. AB - Nine cases of Spigelian hernia occurred and surgically treated in the years 1992 1997 are reported. Spigelian hernias were observed in 5 females and 3 males with mean age of 62.1 years (range 49-70). In 5 cases prosthetic repair has been done with preperitoneal mesh and a very good outcome. In a 61 years old obese female spigelian hernia was bilateral. Her left sided hernia needed an emergency operation for strangulation. Hernia has been repaired by simple suture and recurred early. For the diagnosis of Spigelian hernia it is essential to remember it inside the "Spigelian belt". The satisfactory results obtained at present by prosthetic repair are underlined. PMID- 11059239 TI - [Spigelian hernia: 3 case reports]. AB - Spigelian hernia is an uncommon hernia of the abdominal wall, often not easily diagnosed. Three cases of Spigelian hernia are reported. In the first patient it was an incidental discovery, in the second one hernia was complicated by strangulation of the small bowel, in the third patient hernia was symptomatic without complications. In all the patients, in their previous clinical history, an operation for hernia or other abdominal pathologies was observed. These patients were treated surgically, using direct reconstruction of the abdominal wall in two cases, while a non-absorbable mesh was positioned in the preperitoneal space in the third patient. The anatomopathological and clinical features of Spigelian hernia are described and the problems in diagnosis discussed; in addition, the importance of surgical cure in incidental hernia is underlined. PMID- 11059240 TI - [Septic shock caused by perforation of sigmoid diverticulum into the Retzius space. A case report]. AB - An uncommon complication of the sigmoid diverticulitis personally observed is described. A woman, 59 years old, presented a septic shock caused by perforated sigmoid diverticulum into the space of Retzius. The diagnosis was carried out by an X-ray of the abdomen that suspected the presence of air in the retroperitoneum and then by barium enema showing a perforation of the sigmoid colon. During laparotomy a perforation of sigmoid diverticulum in to the space of Retzius was observed. So the space of Retzius was opened and then pus and watery exudate containing bubbles of gas were found extended also to the retroperitoneum and the right abdomen wall. The patient was submitted to a sigmoid resection; bowel continuity was restored by circular stapled anastomosis and a temporary colostomy was made. Some drainages were placed into the space of Retzius, into the retroperitoneum and into the peritoneal cavity. Nevertheless the patient died because of the septic shock and the postoperative MOF. PMID- 11059241 TI - [Ischemic pathogenesis of acute pancreatitis. A case report]. AB - An ischemic origin of acute pancreatitis has been considered for a long time, at least as an aiding factor: ischemia has a fundamental role in the development of necrotizing pancreatitis from an oedematous one. Shock, heart condition and celiac-mesenteric ischemia can determine the onset of an acute pancreatitis through local ischemic lesions. Personal experience with a case of acute pancreatitis following an intestinal ischemic failure is reported. The duration of ischemia and, in particular, the free radicals formation during the organ reperfusion have been considered as the main pathogenetic factors. In the observed case, the ischemic hypothesis seems to be supported from the lack of other known factors and from the intestinal ischemic failure episode. PMID- 11059242 TI - [Posterior approach in the treatment of sacrococcygeal chordoma: a rare, locally infiltrating, destructive and recurrent tumor. A case report and review of the literature]. AB - Based on personal observation of a case of recurrent chordoma in the sacrococcygeal region four years after the first excision with subtotal resection of the coccyx using a posterior approach, the authors describe the unique characteristics of this rare neoplasm and underline the simplicity of this method of treatment. The surgical technique used to treat the recurrent form also used a posterior approach, with complete resection of the neoplasm including the scar of the previous operation and the last remaining segment of the coccyx. No intra- or postoperative complications were reported. The authors examine the clinical and histological aspects of this neoplasm, concluding that the correct form of treatment must aim both to protect the adjacent anatomic structures and to achieve radical exeresis, as well as achieving a simpler surgical approach unhampered by complications. The authors believe that this approach is simpler than the abdominal-dorsal approach, although this is necessary in the event of intra-abdominal infiltrations. PMID- 11059243 TI - [A case of complete section of the pancreas during closed abdominal trauma]. AB - A review of the literature on pancreatic trauma and its treatment in the light of recent experience is presented. The incidence, mechanism, classification, diagnosis, treatment and complications of pancreatic trauma are discussed. A case of pancreatic transection after blunt abdominal trauma is described. PMID- 11059244 TI - [Treatment of complex anal and rectovaginal fistulas using the endorectal mucosal flap technique]. AB - BACKGROUND: Aim of the study is to evaluate the advantages of the surgical technique for the treatment of complex anal and rectovaginal fistulas by means of an endorectal mucosal flap and the removal of the fistular tract. The surgical removal of a fistular tract can lead to incontinence troubles, mostly for high complex transsphincteric fistulas. The endorectal mucosal flap technique involves the meticulous care of the sphincterial structures and of the mucosa of the anal canal, and is, in personal opinion, the best technique for the treatment of such fistulas. METHODS: The procedure used involves: meticulous study of the anatomic characteristics of the fistula; excision of the secondary opening up to the sphincters level with an elliptic excision of the skin by avoiding keyhole deformities; excision of the primary opening from inside the anal canal, with the interruption of the muscular fibers and their reconstruction; the preparation of an endorectal mucosal flap that is transposed to cover the primary opening of the fistula. 21 patients were treated with this technique, 17 were complex anal fistulas, two were associated to Crohn's disease, and of two rectovaginal fistulas, one was found in a patient with Crohn's disease. RESULTS: Recurrences were found in five patients (24%), after a median follow-up of 20 months. The delay of fistular healing, over a month, was found in 50% of the cases. No patient had continence troubles. CONCLUSIONS: The mucosal flap technique is a simple procedure, adequate in preserving the sphincteric function and in preventing deformities of the anal canal. However, it has not demonstrated clear benefits in terms of recurrences compared to other techniques, but it is highly considered for the treatment of complex anal fistulas, in terms of continence and for the earlier and better healing of the wounds and for the better tolerance of the patient. PMID- 11059245 TI - [Photography methods in general and esthetic surgery: clinical significance and impact on surgical performance]. AB - The authors outline and emphasize the use of Polaroid technique in the surgical practice, either in emergency room (pre-, intra- and postoperative photography) or in plastic-reconstructive surgery, where informed consent is needed on the basis of surgical plan. Polaroid photos are suitable to be enclosed in the patient record as well as in the legal documentation. The Polaroid Macro-5SLR camera is specifically useful in the medical practice to improve the doctor patient relationship. PMID- 11059246 TI - [Legislative, medicolegal and economic aspects of day hospitals]. AB - After a short historical review and having dwelled on the nomenclature and legislative framework, the authors describe the medicolegal implications of the day hospital. They describe international experience within the various disciplines, with reference to economic aspects in terms of costs and the increased efficacy of these structures, the quality of service and government policies, in particular in industrialised countries. The authors conclude that day hospitals present considerable advantages both in economic terms and for patients. However, the organisation of the system needs to be improved and, where possible, procedures standardised. Bearing in mind the importance of costs to modern medicine, it is impossible not to take economic aspects into account, while also improving the quality of relations between doctors, patients and the health system. PMID- 11059247 TI - Systems analyst. PMID- 11059248 TI - The doctor is getting out. PMID- 11059249 TI - Physicians who write. PMID- 11059250 TI - My wife, my hero. PMID- 11059251 TI - Election 2000. What your vote may mean for health care policy. PMID- 11059252 TI - Antibiotic bacterial resistance in ambulatory patients. AB - BACKGROUND: This study evaluates trends in antibiotic resistance in patients who were treated in an ambulatory setting. METHODS: The authors compiled the data from all lower respiratory track(sputum) cultures collected from ambulatory patients who visited the Olmsted Medical Center and Mayo Clinic between 1985 and 1998. Cultured organisms were identified, and Minimal Inhibitory Concentration (MIC) values were presented and categorized as susceptible, intermediate, or resistant based on the National Committee for Clinical Laboratory Standards (NCCLS) guidelines for MIC and antibiotic susceptibility. RESULTS: 4,297 potentially pathogenic organisms were obtained from sputum cultures for 1,921 patients. The most discernible changes in antibiotic resistance appeared to be in cultures positive for Pseudomonas aeruginosa. A trend toward increasing resistance of isolates of Streptococcus pneumoniae to beta-lactam drugs was observed in a portion of the population. An emerging intermediate susceptibility among isolates of Klebsiella pneumoniae and Pseudoumonas species was noted. CONCLUSIONS: Trends in antimicrobial resistance of respiratory pathogens from ambulatory patients are less clear than those from hospitalized patients, but must be monitored because of the high percentage of ambulatory patients who receive empirical therapies. Trends in intermediate susceptibility patterns may help reveal emerging antimicrobial resistance. PMID- 11059253 TI - Medicare managed care reimbursement. Unjust yet constitutional. PMID- 11059254 TI - [The readiness to communication and self-presentation as chosen aspects of social activity in depressive patients]. AB - Willingness to communicate and self-presentation were examined in 68 patients with depression. The results of the patients were compared with a control group of healthy individuals. The results indicated changes in social life of patients, especially in interpersonal communication. Depressive patients showed significantly lower level of willingness to communicate than healthy persons. The self-presentation letters of patients mostly included information about the disease, negative feelings and thoughts connected with depression and negative self-description. PMID- 11059255 TI - [Sexual functioning of patients with major depression. Pilot study]. AB - Disturbances in sexual functioning in patients with endogenous mental disorders are frequently reported. The aim of the study was to estimate the occurrence of sexual disfunctions in patients hospitalised for major depression (unipolar affective disorder). The investigations covered 53 persons remaining in stable partnership. Apart from the assessment of psychological state, depressive symptoms and the course of treatment, we analysed the quality of relations with the partner, sexual satisfaction, willingness to and frequency of sexual intercourse, satisfaction with oneself as a sexual partner. The obtained results indicate that sexual problems must not be neglected in treatment of depressive patients. PMID- 11059256 TI - [The analysis of diagnoses and pharmacological treatment of patients on disability pension due to depression]. AB - The aim of the study was to assess the diagnoses and pharmacological treatment in 80 patients with symptoms of depression in reference to the psychiatric examination due to disability pension proceedings. Diagnoses were often not in accordance with the 10th revision of the International Statistical Classification of Diseases and Related Health Problems (ICD-10). Twenty two per cent of outpatients were treated using benzodiazepines only. In the group of antidepressants, mianserin, fluoxetine and doxepine were used most often (in 12%, 12% and 11% of outpatients respectively). Inpatients were usually treated with doxepine (24%) and mianserin (18%). As many as 15% of patients received benzodiazapines as the only drug during the hospitalisation. In contrast to the newer generation of drugs (moclobemide, fluoxetine), tricyclic antidepressants were usually used in low, not therapeutic doses. Further studies on the quality of treatment of depression in Poland, based on the more representative groups are necessary. PMID- 11059257 TI - [New implications for mental disorders treatment: pharmacogenetic studies of dopaminergic and serotonergic neurotransmission systems]. AB - The aim of this article is to summarize the present state of pharmacogenetical knowledge compiled and based on various publications in this matter. Thanks to developing cloning and DNA-analysis techniques it is possible to analyze numerous proteins synthetized in the central nervous system. Several polymorphism sites in coding genes have already been located--first of all for some genes coding receptors linked to protein G, especially for dopamine and serotonine receptors. Some of mutations may influence the primary structure of receptor protein and in that way be responsible for alteration of receptors functioning. This is likely to be the reason for the difference in reaction to drugs in many patients. A couple of trials dealing with patient's response to neuroleptics in correlation with receptor genes polymorphism have already been completed. The results are promising. The assumption that inventing new drugs should be correlated with collecting DNA samples from patients to evaluate further pharmacogenetical linkage seems to be essential. PMID- 11059258 TI - [Pharmacokinetics of second generation antidepressants]. AB - This paper presents the pharmacokinetic properties of newer second generation antidepressants, with an emphasis on the main metabolic pathways of drugs and metabolites as well as their effects on the cytochrome P450 enzymes. Except for Milnacipran all newer antidepressants are biotransformed in the presence of at least one cytochrome P450 izoenzyme. Drugs and their metabolites may be the substrates or inhibitors of cytochromes P450. The knowledge of drugs' metabolic pathways as well as the knowledge of substrates and inhibitors of izoenzymes assists in choice of a proper drug and its dose. It is particularly important in case of polypharmacotherapy when there is a high risk of adverse events. It is also important to remember about genetic polymorphism of some cytochrome P450 izoenzymes (CYP2D6, CYP2C19) that underlies the marked inpatient variability in drug metabolism. PMID- 11059259 TI - [Effect of therapeutic drug monitoring of amitriptyline and genotyping on efficacy and safety of depression therapy]. AB - Modern pharmacotherapy is based on precise adjustment of a dosage schedule to individual requirements of patient. Therapeutic drug monitoring is a method that allows for a more effective treatment approach, especially in the case of a narrow therapeutic index of a drug. Tricyclic antidepressant drugs are characterised by narrow therapeutic index as well as relationship between serum drug concentration and side effects. It was demonstrated that interindividual variability of blood concentrations of tricyclic antidepressant drugs is related to genetic polymorphism of oxidating enzymes participating in metabolism of these drugs. The aim of the study was to estimate the impact of therapeutic drug monitoring of tricyclic antidepressant drugs as well as genotyping on efficacy and safety of endogenous depression therapy. The study included 9 patients with established diagnosis of endogenous depression. Blood serum concentrations of amitryptyline was measured by fluorescence polarisation immune assay (FPIA, Abbott system). Genotype of cytochrome P450 isoenzyme CYP2D6 was determined using PCR-RFLP method. It was demonstrated that monitoring therapy of tricyclic antidepressant drugs in combination with determination of the genotype seems to be more safe and effective. Monitoring therapy and genotyping may be less expensive than the costs of prolonged hospitalisation and risk of side effects. PMID- 11059260 TI - [The involvement of 5-HT1a serotonin receptors in the pathophysiology and pharmacotherapy of schizophrenia]. AB - Many studies of schizophrenic brains indicate the dysfunction of dopamine and glutamate systems in the prefrontal and frontal cortex. It seems that better understanding of mechanisms regulating functions of these neuronal cortical systems could contribute to creation of new drugs acting in the cortex selectively. This might be profitable in cognition of dysfunction and negative symptoms in schizophrenia. This article presents preclinical data concerning the role of 5-HT1A serotonin receptors in the modulation of cortical dopamine system and in psychotomimetic effects of non-competitive NMDA receptor antagonists. Neurochemical studies have shown that 5-HT1A receptor agonists increase dopamine release in the rat prefrontal cortex on the one hand, and they inhibit the augmentation of dopamine release induced by stress or amphetamine, on the other. However, the increase of dopamine release induced by non-competitive NMDA receptor antagonists is blocked by 5-HT1A receptor antagonists. Blockade of 5 HT1A receptors seems to be important also in reduction of most psychotomimetic effects induced by non-competitive NMDA antagonists both involving (locomotor hyperactivity, working memory impair) and not involving (sensorimotor gating deficits) dopamine mechanism. Thus, binding with 5-HT1A receptors can be an important site for the regulation of cortical dopamine system, both in physiological conditions and in disregulation of the system induced by stress, psychostimulants or psychotomimetics. On the other hand, 5-HT1A receptors modulate most of psychotomimetic effects of non-competitive NMDA receptor antagonists. The above results of preclinical investigations indicate that 5-HT1A receptor can be involved in the pathology of schizophrenia, what is partly confirmed by clinical postmortem studies of schizophrenic brains. These studies showed the increase of 5-HT1A receptor density in prefrontal and frontal cortex in schizophrenic brains. It also seems that 5-HT1A receptors might be a good target for the antipsychotic drugs. Although the clinical studies have demonstrated controversial data, maybe further studies using substances with selectivity to 5-HT1A receptors would help to determine more precisely the role of these receptors in pathology and pharmacotherapy of schizophrenia. PMID- 11059261 TI - [Perspectives of therapy of Alzheimer's disease]. AB - Alzheimer's disease is the most common cause of memory disruption in elderly people. The main pathogenic factor of the disease is beta-amyloid protein, which may cause toxic damage of neurones. Other suggested pathogenic factors include an inflammatory process around the senile plaques, apoptosis and necrotic death of neurones, and, in consequence, changes in functioning of neurotransmitter systems. In this article the authors present the main directions in pharmacotherapy of Alzheimer's disease: causal therapy, which prevents the neurodegenerative changes and slows down the pathogenetic process, and symptomatic therapy. The aim of symptomatic therapy is to reduce memory disruption and psychiatric symptoms associated with the disease. Positive influence on cognitive processes is exerted by cholinergic drugs, e.g. the actually used inhibitors of acetylcholinesterase (rivastigmine, donepezil), the nootropic agents (piracetam, nefiracetam) and extracts of Gingko biloba. For treatment of the disease accompanying psychiatric symptoms (anxiety, depression, hallucinations, sleepness) the drugs with minimal influence on cognitive processes are recommended. Attempts at causal therapy are focussed on searching for the substances that can prevent the formation and toxicity of beta-amyloid (droloksifen, estrogens, agonists of muscarinic receptors M1), the cytotoxic influence of excitatory aminoacids (memantine, lamotrigine), calcium (nimodipine) and free radicals (selegiline, alpha-tocoferol), and the development of inflammatory process (non-steroidal antiinflammatory drugs). The new target of research is correction of deficits of nerve growth factor and neurotransmitters by intracerebral implantation of modified fibroblasts. Another way is prevention of the formation of amyloid plaques using appropriate antisense oligonucleotides. PMID- 11059262 TI - [The effect of zolpidem on EEG activity in falling asleep process]. AB - The effect of zolpidem on the brain bioelectric activity in falling asleep process of 13 healthy volunteers (8 women, 5 men), aged 20-24 years, was investigated. Its effect on sleep latency was studied. Quantitative analysis of sleep spindles occurrence was realized. It caused significant increase of sleep spindles activity in the first hour after its application (p < 0.001) and had influence on reduction of sleep latency in comparison with placebo. Our findings confirm the hypnotic effect of zolpidem. PMID- 11059263 TI - [The influence of whole body cryotherapy on mental health]. AB - The paper presents a little known issue about the influence of wholebody cryotherapy on mental health. Observations of patients' behaviour after passing the cryogenic chamber leads to an interesting hypothesis. Short exposition to extreme cold has doubtless a profitable influence on man's frame of mind. Immediately after passing the cryogenic chamber, apart from the well known analgetic effect, we detect changes in patients' mental state such as improvement of mood, deep relaxation, freshening up, consolation, euphoria. This unusual state lasts for a long time after ending the cycle of cryotherapy. Different mechanisms of this effect are considered. New possibilities of this method have been presented. Durability of such an advantageous phenomenon are investigated in our research centre in Wroclaw. PMID- 11059264 TI - [Transcranial magnetic stimulation versus electroconvulsive shocks: neuroanatomical investigations in rats]. AB - Since the time of introducing ECT to the clinical practice, the method always raised questions regarding possibility that the current running through structures of a brain may evoke structural changes and, as a result, evoke convulsive attacks. Pathological changes (swelling, gliosis, atrophy, necrosis) were observed most often after "mega"--schemes including series of several to several hundred ECT treatments. Regime used nowadays including only 8-12 ECT sessions seems to be entirely safe. There are, however, only a few experimental works dealing with this problem. In 1992 research started on a new neurophysiological technique--transcranial magnetic stimulation (TMS) in depression. The advantage of this method is that is does not seem to evoke convulsive attacks. Prolonged rapid rate TMS (rTMS) seems to be particularly efficient in treatment of depression. Despite thousands of works describing various functional effects of TMS, there are obviously no researches on structural effects of the technique. In the case of experimental research on animals a few works were published and their results seem to be ambiguous. We have examined the influence of prolonged repetitive rTMS (B = 1.4 T, t = 5.5 min, f = 30 Hz), and standard ECT (I = 150 mA, t = 0.5 s, f = 50 Hz) on the structure of brain tissue in rats. Both groups of animals (n = 10) received 12 stimulation sessions. After the treatment the animals were routinely processed for electron microscopy (EM) and for light microscopy (LM). Our investigations suggest that the technique of ECT shows a considerable neurotoxic potential. In comparison to ECT--the rTMS method seems to be more safe. PMID- 11059266 TI - Society of Psychophysiological Research 40th annual meeting. October 18-22, 2000. San Diego, California, USA. Abstracts. PMID- 11059265 TI - [Internet telediagnosis and teletherapy which should not do harm]. PMID- 11059267 TI - [Molecular modification and artificial evolution of microbial enzymes]. AB - There exists a remarkably progress in molecular modification and artificial evolution of microbial enzymes in recent years. Error-prone polymerase chain reaction and DNA shuffling have been developed and successfully applied and a lot of startling achievements have been obtained in artificial evolution of enzymes. PMID- 11059268 TI - [Bioconversion of hemicellulose hydrolysates for xylitol production]. AB - Xylitol has attracted much attention because of its many applications in the food, medicine and chemical industries. However the use has been limited by its high price. This coast is a result of the extensive purification steps needed for the preparation of a pure xylose solution, which is essential for the chemical process. The fermentative process of xylitol is an interesting alternative to conventional chemical process, since it does not require initial xylose purification. The present review describes the advantage of xylitol production by fermentation, xylitol-producting microorganisms, metabolic pathway of xylose in yeasts, detoxification of hemicellulose hydrolysates and fermentative conditions affecting xylitol production. PMID- 11059269 TI - [Synonymous codon usage in Pichia pastoris]. AB - According to the synonymous codons used in 28 open reading frames from Pichia pastoris, the codon usage in this species was calculated and 19 codons have been inferred to be its optimal codons. The results show that pattern of the codon usage in P. pastoris is similar to that in S. cerevisiae and in K. lactis except for the synonymous codon of glutamic acid, which may be the special bias of P. pastoris. PMID- 11059270 TI - [Expression and characterization of two kinds of recombinant snake neurotoxins]. AB - The cDNA encoding the precursor of cobrotoxin was cloned from the venom gland of the Chinese continental cobra (Naja naja atra) by RT-PCR. Its deduced amino acid sequence analysis showed that the mature protein was identical to that identified from the Taiwan cobra (Naja naja atra) by protein sequencing technique. The cDNA encoding the mature protein was then subcloned into the expression vector pMAL P2. The gene of CM11, which was formed by ligation of the fragments of the synthetic oligonucleotides, was also cloned into the expression vector pMAL-P2. After induction of IPTG, both of the neurotoxins were overexpressed as soluble fusion proteins which were confirmed by SDS-PAGE and western blotting. The expressed fusion proteins was purified by sepharose 6B-amylose affinity chromatography and DEAE-sepharose FF chromatography. Both of the recombinant proteins achieved after digestion by factor Xa showed the in vivo toxicity. PMID- 11059271 TI - [Studies on the expression of the recombinant human GM-CSF/IL-3 fusion protein]. AB - A human granulocyte-macrophage colony stimulating factor (GM-CSF)/interleukin 3(IL-3) fusion gene with a short linker between the GM-CSF and IL-3 gene has been successfully constructed and expressed in E. coli under the control of T7 promoter. The recombinant fusion protein was expressed as inclusion bodies after the IPTG induction. The yield of the GM-CSF/IL-3 fusion protein was over 30% of the total cellular proteins. Western-blotting results showed that the fusion protein could specifically combined with GM-CSF antibody and IL-3 antibody. The biological activity was detected by the GM-CSF and IL-3 dependent cell line TF-1. After solubilizing with 8 mol/L urea and renaturing with dialysis against Tris. HCl solution, the refolded fusion protein showed obvious activities to maintain the growth of TF-1 cell. PMID- 11059272 TI - [Cloning of a full length cDNA of human thrombopoietin receptor c-Mpl and construction of engineered cells that stably express c-mpl]. AB - A full length cDNA fragment encoding for human thrombopoietin receptor c-Mpl has been amplified by RT-PCR from the total RNA of human HEL cells. The complete sequence of the cloned cDNA was determined and is identical to that previously reported. Then the fragment was subcloned into the mammalian expression vector pcDNA3 and the resulting plasmid is designated as pcMPL. K562 cells, which do not express c-mpl, were transfected with pcMPL and pcDNA3, respectively. The transformants were selected with G418 and then tested by Northern and Southern blotting. A group of engineered cell lines stably expressing c-mpl have been obtained, which will facilitate further research on the signaling mediated by c Mpl. PMID- 11059273 TI - [Utilization of a microsatellite marker to identify rice blast resistance gene in two segregating populations]. AB - The microsatellite (SSLP) marker RM262, which is tightly linked to the blast resistance gene Pi-d(t) in rice, provides means to conduct marker-aided selection in a rice breeding program. The objective of this study was to investigate the ability of this marker to select the blast resistance gene, Pi-d(t), in two crosses for rice blast resistance breeding. The products with amplified the microsatellite primers were polymorphic between the three varieties examined. To examine the power of the identified microsatellite marker in predicting the Pi d(t) locus, we determined the genotypes of the two F2 populations at the Pi-d(t) locus by performing progeny testing for the disease respones. The results indicated an accuracy of more than 98% in identifying the resistant plants in both populations. Therefore the microsatellite marker can be utilized in marker assisted selection and breeding for new varieties with blast resistance. PMID- 11059274 TI - [Cloning cDNA of extracellular domain of human erythropoietin receptor and its expression in Escherichia coli]. AB - Human erythropoietin receptor (hEpoR) plays an important role in regulating the red blood cell production by promoting the proliferation and differentiation of RBC from erythroid precursors. hEpoR is a transmembrane protein, and its extrocellular domain (sEpoR) is of great importance in Epo signal transduction pathway. We cloned the gene of sEpoR by RT-PCR from the total RNA of human fetal liver and expressed it in E. coli after insertion of the gene in the expression vector pBV220. The cloned gene was confirmed by sequencing analysis and gene product was confirmed by both Western blot and its first 11 amino acid residues sequence of the N-terminal. In vitro bioassay showed that the purified gene product can repress the growth of TF-cells in the presence of Epo. PMID- 11059275 TI - [Induction of protective immune response in mice and rhesus monkeys by immunization with fusion protein of cholera toxin B subunit and multiples of Plasmodium falciparum]. AB - Recombinant fusion protein of cholera toxin B subunit (CTB) and poly-valent protective epitopes of plasmodium falciparum was given to i.m. to C57BL/6j mice and rhesus monkeys three times. In rhesus monkeys, high level of antibodies for CTB (1:6400) and malaria epitopes (1:3200) amtobpdoes were elicited as well as the specific CTL activity for P. plasmodium. After the mice were challenged with sporozoites of P. yeolli, about 50% of them were protected from the patent infection. A blood-stage challenge with 10(8) of P. cynomolgi parasite were given to rhesus monkeys, which showed that two animals in control group were patent infection for at least 30 days, in contrast, the two animals immunized were recovered respectively at the day of 11 and 15 after challenges. The results suggested that cholera toxin acts as an effective adjuvent in the development of malaria vaccine. PMID- 11059276 TI - [Functional and structural study of the prokaryotic enhancer-like element VV16 from vaccinia virus genome]. AB - An enhancer-like element VV16 from Vaccinia virus genome DNA was obtained by using the plasmid with CAT reporter gene. Sequence analysis showed the element of 112 bp is a part of the DNA-dependent RNA polymerase, polyA polymerase and DNA polymerase (RPO30 gene). It contains 4 AT-rich regions. Detection of beta galactosidase activity showed that VV16 in the positive direction can increase the activity 9.0 times and VV16 in the negative direction can increase 4.1 times. The RNA dot blotting confirmed the enhancing activity of the element are on the transcription level. DNA deletion experiment indicated the sequences of 10 bp at the 5' end and 12 bp at the 3' end in the element are important to its function and the sequence from nt76 to nt82 is essential to its activity. PMID- 11059277 TI - [Site-directed mutagenesis and effects on the enzymatic properties of subtilisin E]. AB - Site-directed mutagenesis was used to investigate the effects of S221C/P225A, N118S/S221C/P225A, D60N/N118S/S221C/P225A and Q103R/N118S/S221C/P225A mutations on the properties of Subtilisin E. It was found that S221C/P225A mutant is 73,000 fold decreased in amidase activity than subtilisin E and 3-fold increased than subtiligase in the ratio of esterase/amidase; N118S/S221C/P225A mutant has 3.6 fold and 15-fold decreased in amidase and esterase activity respectively and as a result, it has a 4-fold lower in the ratio of amidase/esterase than S221C/P225A mutant; Although it has no effect on the esterase activity, D60N/N118S/S221C/P225A mutant enhanced its ratio of amidase/esterase by 15 fold, 3.3-fold and 10.3 fold compared to N118S/S221C/P225A mutant, S221C/P225A mutant and subtiligase respectively; Q103R/N118S/S221C/P225A mutant, however, has a 5 fold enhanced in the amidase activity and 55-fold and 1000-fold decrease in the esterase activity and the ratio of esterase/amidase compared to N118S/S221C/P225A. PMID- 11059278 TI - [Purification and characterization of recombinant Aeromonas punctata prolyl endopeptidase]. AB - The study of down-stream techniques of recombinant Aeromonas punctata prolyl endopeptidase (apPEP) was presented here. High cell-density fermentation of E. coli BL21/pKKH-PEP in NBS BioFlo 3000 5 L fermentor was achieved, the final cell density was 22.5 g (DCW)/L after 14 h cultivation, the yield of apPEP expressed in soluble protein was 3.0 g per litter broth. After sonication, the supernatant of free cell extract was purified by ammonium sulfate fractionation, High performance Q sepharose FF, Phenyl sepharose 6 FF, the purity of apPEP reached 96%, enzyme specific activity was 65.5 u/mg, apPEP yield reached 0.86 g/L broth. Total recovery of enzyme protein was 8.2%, actviity recovery was 24.4%. The molecular weight of apPEP was 76,464 +/- 30 Da measured by MS, N terminus amino acids sequence consistent with that deduced from DNA sequence. pI 6.0, which was similar with PEP from Aeromonas hydrophila. PMID- 11059279 TI - [Expression of prourokinase in different mammalian cells]. AB - Comparison studies of recombinant prourokinase(pro-UK) in various host cells, and expression vectors were carried out. Expression levels of vectors constructed in this study in different cell lines were compared. Mammalian cells expressing pro UK were established. The levels of pro-UK expression in recombinant Namalwa, Vero and Sp2/0 cells are 200, 12.5 and 50 IU/(10(6) cells 24 h). pro-UK purities separated by immunoaffinity chromatograph are above 90%. Immunoabsorbent assay showed the ratio of pro-UK in CHO cells is lower than that from Vero and Namalwa cells. This study provide new host cells for pro-UK production. PMID- 11059280 TI - [cDNA cloning, fusion expression in Escherichia coli and activity assay of hIL 11]. AB - Human Interleukin-11 (hIL-11) is a multifunctional cytokine which plays an important role in regulating the proliferation and differentiation of cells in the hematopoeitic, lymphoid system etc. To obtain the IL-11 cDNA, a primary culture of Chinese fetal lung fibroblast was prepared from fresh tissue. Then the human IL-11 cDNA without the N-terminal signal peptide sequence was cloned by RT PCR from the cells induced by PMA. The sequence indicated that there are three bases different from those previously reported, but with no change of the amino acids. The cDNA was inserted into the 3' end of trxA gene in thioredoxin gene fusion expression system pTRXFUS to construct the trxA and hIL-11 fusion expression vector, and expressed in E. coli. The fusion hIL-11 accounts for more than 20% of the total bacteria proteins. The expression product is present in soluble forms and has the full biological and immunological activities. PMID- 11059281 TI - [Application of intermittent-feeding of growth-limiting nutrients in suspension culture of insect cells(Sf21)]. AB - On the basis of the growth and metabolism behavior inherent in suspended Sf21 insect cells, the intermittent feeding of growth-limiting nutrients(glucose and protein hydrolysates) was employed in the regulation of cellular growth during the cultivation by using the residual glucose concentration as a reference point. It was shown that as compared with the batch cultivation, the intermittent feeding of growth-limiting nutrients effectively prolonged the growth and stationary phase for Sf21 insect cells grown in two representative culture medium(TC-100 and IPL-41). The maximum cell density was increased from 3.0 x 10(6) cells/ml to 6.5 x 10(6) cells/mL in TC-100 medium, and in IPL-41 medium, the maximum cell density was increased from 7.05 x 10(6) cells/mL to 9.0 x 10(6) cells/mL. As the defined nutrient solution was used for feeding in lieu of the complicated and expensive base medium, the technique would find prospects in large scale high-density cultivation of insect cells. PMID- 11059282 TI - [Studies on lipase-catalyzed synthesis of L-ascorbyl palmitate in non-aqueous phase]. AB - The investigation on the reaction media and lipases (NOVO435, MML, LIPOLASE, PPL) for enzymatic synthesis of L-ascorbyl palmitate as well as the factors effecting initial rate of synthesis reaction (rotation speed, temperature, water content, enzyme concentration, and substrate concentration) is presented. Among these investigated solvents and lipases, amyl alcohol and NOVO435 is the optimum pair. The reaction conditions have been optimized: 200 r/min, 55 degrees C, water content = 0, the amount of enzyme = 12.5% of the substrate's quantity. PMID- 11059283 TI - [Studies on simultaneous production of chitosanase and chitosan degradation in situ with Trichoderma reesei in convoluted fibrous bed bioreactor]. AB - The cells of Trichoderma reesei were immobilized on a roll of porous polyurethane foam sheet and packed in a bubble column bioreactor for simultaneous production of chitosanase and degradation of chitosan in situ. The average degree of polymerization could be regulated by reaction time. Under the repeated-batch process with 2% soluble chitosan at pH4.8, 28 degrees C, the activity of chitosanase for each batch was above 0.15 IU/mL, the average yield of reducing sugar as D-glucosamine reached 73%. The novel immobilized bioreactor system run stably and effectively in the successive 10 batches lasting 30 days without notable change in the activity and productivity. PMID- 11059284 TI - [Growth and metabolism of hybridoma cells cultured in flask and spinner bottles]. AB - The growth and metabolism of WuT3 hybridoma cells were very different when cells were cultured in flask and spinner bottles. In the flask bottles, a longer culture duration and the higher cell density were met, whereas in the spinner bottles the more vigorous cell metabolism took place, resulting in that either the specific consumption rates of glucose and amino acids or the specific production rates of lactate, ammonia and alanine were higher. PMID- 11059285 TI - [Studies of the mechanism of 7-epi-taxol converted to taxol from Taxus extract catalyzed with Al2O3]. AB - Under the catalysis of Al2O3, 7-epi-taxol in Taxus yunnanensis extract could convert to taxol. The factors including the concentration of CH3OH in the mobile phase, the type of Al2O3 and the reaction time could influence the isomeric reaction. The catalysis mechanism fo Al2O3 was investigated. The isomerization of 7-epi-taxol was under the cooperation of the Lewis souci activity core and the basic activity core in the surface of Al2O3 at room temperature. The type and strength of activity core controlled the procedure of reaction, and the suitable content of CH3OH in the mobile phase would accelerate the reaction. PMID- 11059286 TI - [Enhancement of fermentative glycerol yield with heat shock treatment]. AB - A heat shock treatment was studied in glycerol fermentation of osmotolerant yeast Candida krusei. The experiment results showed that the optimal temperature and duration time for heat shock is 45 degrees C and 30 min respectively, and the optimal start time of the treatment is at the mid term in exponential growth stage. With such treatment, glycerol yield was enhanced greatly, while no significant effects on both cell growth and glucose consumption were observed. PMID- 11059287 TI - [Production of u-PA with rCHO cell culture on porous microcarriers in serum-free growth medium]. AB - A novel technique was developed to deal with apoptosis in large-scale animal cell culture. By means of replacing part of Cytopore porous microcarriers at regular intervals, a rCHO cell line, which produces urokinase-type plasminogen activitor(u-PA), was cultivated continuously with serum-free medium in a 30 L stirred tank for 91 days. The cell density was maintained at (1.3-2.6) x 10(7)/mL, and > 90% of cells was viable. In order to reduce the effect of cell density on cell growth and expression, a cyclic pressure oscillation was exerted on a 7.5 L reactor headspace to enhance cell expression at high cell density to a certain extent. During the 67 days of medium-replacement culture, the maximal cell density reached 2.64 x 10(7)/mL, and cell viability was always kept above 95% when combined with microcarrier-replacement. Compare to control culture, culture with cyclic pressure oscillation could enhance cell expression level and reduce the ratio of glucose metabolized anaerobically to produce lactate. With four-step purification process, about 80 g u-PA(approximately 90% scu-PA) was recovered from approximately 2100 liters supernatant which contained approximately 135 g u-PA. PMID- 11059288 TI - [Study on continuous synthesis of galacto-oligosaccharide by immobilized Bacillus stearothermophilus]. AB - The galacto-oligosaccharide was synthesized continuously by immobilized Bacillus stearothermophilu producing beta-galactosidase in fibrous bed reactor. The effect of substrate concentration, pH, reaction temperature and retention time on production of GOS was investigated. The optimal reaction conditions were determined. Substrate concentration were 450 g/L; Reaction temperature was 55 degrees C; pH was 7.0; Residence time was 100 min. The product yield reached up to 50.7%. GOS synthesis was promoted by feeding 1.5% D-glactose after 24 h. The immobilized cell reactor can work stably for 120 h. PMID- 11059289 TI - [Nutrition condition of hyaluronic acid fermentation with Streptococcus zooepidemicus]. AB - Based on the analysis of metabolic pathway Streptococcous zooepidemicus for hyaluronic acid (HA) synthesis, nucleotide, especially uracil, was considered to be important to cell growth and metabolism. When 0.005 g.L-1 uracil added in the media in which yeast extract as complex nitrogen source, cell growth and HA production were increased by 32% and 34% respectively. From analysis of amino acid in fermentation process, it was show that arginine(Arg) was needed for cell metabolism, and concentration of free Arg maintained at 0 g.L-1 in fermentation process, which was proposed to limit cell growth and HA production. By shake flask experiment HA concentration reached 0.510 g.L-1 when 0.06 g.L-1 Arg added, in the fermentation with 2.5 L fermentor, when uracil 0.005 g.L-1 and Arg 0.06 g.L-1 were added, the rate of cell growth increased, maximum of specific growth rate, concentration of HA and HA molecular weight reached 0.67 h-1, 5.2 g.L-1 and 2.15 x 10(6) Da from 0.54 h-1, 4.2 g.L-1, 2.0 x 10(6) Da, respectively. PMID- 11059290 TI - [High-level expression of BYDV GAV coat protein gene in Escherichia coli]. AB - The IPTG-inducible expression vector containing the BYDV GAV coat protein gene was constructed and transferred into E. coli BL21(DE3). High-level expression of the specific protein was achieved by IPTG induction. The results of SDS-PAGE and Western blotting show that the expression product which accumulates 19.5% of the total cellular proteins estimated by scanning is 24 kD BYDV GAV coat protein plus eleven amino acids of pET-5a. PMID- 11059291 TI - [A novel rabbit endothelin B receptor gene: cloning and sequence analysis]. AB - Endothelin(ET) is the most potent mammalian vasoconstrictor identified to data. As a pathogenic factor, ET is involved in the genesis of many diseases. In this study, a pair of primers was designed and synthesized according to the human ETB receptor gene (hETBR) sequence. A 394 bp of DNA fragment was amplified by polymerase chain reaction(PCR) and labeled with alpha-32P-CTP using Random Primer Labeling method. With this probe, rabbit lung cDNA library was screened by in situ hybridization and 11 positive clones were identified. Sequencing result showed that a complete reading frame of rabbit ETB receptor(rETBR) cDNA could be produced from three positive clones of eleven. By a series of subcloning, a recombinant plasmid including the 1326 bp of rETBR coding sequences, named pBlu Script-rETBR, was constructed. The deduced amino acid sequence indicated that the rETBR is 441 residues in length, with an expected molecular mass of approximately 49.44 kD. N-terminal 18 residues is the potential signal peptide (Score = 11.11) and therefore the molecular mass of mature rETBR is 47.65 kD with 423 amino acid residues. Analysis of the rETBR hydropathy profile indicates the presence of seven hydrophobic regions, putative transmembrane domains. Potential N glycosylation sites are the 60th and the 118th. The structure exhibits a significant sequence and topographical similarity with G protein-coupled receptors. PMID- 11059292 TI - [cDNA cloning, sequence analysis of trehalose-6-phosphate synthase gene from Saccharomyces cerevisiae AS2.1416]. AB - The trehalose-6-phosphate synthase gene tps1 was amplified from yeast S. cerevisiae AS2.1416 cDNA library by polymerase chain reaction. This 1.5 kb fragment was cloned into PstI and BamHI sites of pGEM-T easy vector and the sequence of the gene indicated the cloned tps1 gene contained 1485 nucleotides encoding for 495 amino acid and shared a sequence homology of 99.6% with that from S. cerevisiae S288C. PMID- 11059293 TI - [Fusion expression of cecropin X including the cleavage of FXa in Escherichia coli]. AB - PCR method was used to introduce the code sequence of Factor Xa cleavage site to the 5' end of cecropin CMIV mutant gene X, then the gene was cloned into the expression vector pGEX-KG, and was highly expressed in E. coli BL21 by IPTG induction. The fusion protein was purified by affinity-chromatography and was cleaved by Factor Xa. Cecropin X with antibacterial activity was obtained after purified by ion-exchange chromatography. PMID- 11059294 TI - [Importance of a wide perspective on meta-analysis. It may be of crucial significance for the patients]. PMID- 11059295 TI - [MEDLINE in the age of Internet]. AB - The use MEDLINE and related resources of literature information has increased significantly with the increased availability and speed of the Internet. However, literature information is presented in many different ways via the Internet and search facilities and web addresses are frequently changed. Many physicians and medical researchers find it difficult to keep track of what is available and many lack skills in effective searching. The present paper presents an overview of the MEDLINE database and related databases available from the National Library of Medicine and searchable through PubMed and Internet Greatfull Med. We discuss strategies for searching MEDLINE and the coverage of MEDLINE related to that of related databases from the NLM and other sources. PMID- 11059296 TI - [Online publishing in the Internet age]. AB - The availability of full text medical journal articles is rapidly increasing with the increased availability of the Internet. The potentials of the new technology present researchers, publishers, and librarians with new problems and challenges. Some resources are made available free of charge, whereas others are distributed as parts of large licences negotiated between publishers and consortia of research libraries. How can researchers maintain an overview of the constantly changing resources? How can libraries cope with tasks rapidly redefined by the technology? And how can publishers survive when production and distribution of literature information, including the handling of peer reviewing, might just as well be performed by the researchers themselves or their organisations? The present paper presents some of the resources available and discusses both national and international projects and activities that deal with these questions. PMID- 11059297 TI - [Acute pancreatitis]. AB - Acute pancreatitis is a common disease with significant morbidity and mortality. Despite recent improvements in our understanding of the disease process and the development of a range of supportive measures, today's treatment approaches are still less than ideal. Here I present some of the key points from generally accepted guidelines for the management of acute pancreatitis, including the operational classification system approved in Atlanta, USA, 1992. Moreover, the diagnostic value of magnetic resonance as well as new approaches to treatment, including data on prophylactic antibiotics, selective digestive decontamination, and enteral nutrition in acute pancreatitis, are discussed in more detail. PMID- 11059298 TI - [Treatment of HIV infections with antiretroviral drugs and recombinant interleukin-2]. AB - The clinical effect of combination antiretroviral therapy against HIV-infection is indisputable, but the current treatment does not produce complete immuno restitution. Many HIV-positive patients change treatment, because of side effects and virological failure. Owing to the limited number of treatment combinations, supplementary treatment is greatly needed. Intermittent subcutaneous rIL-2 treatment plus antiretroviral combination therapy results in a selective and long lasting induction of CD4+ cells in 70-80% of HIV-patients and lowers the amount of replication competent virus in blood and lymph nodes. The expanded cell population consists of both naive cells and memory cells with the ability to respond to antigenic stimulation. It is not known whether the rise in the number of CD4+ cells reflects a better clinical outcome. This question is currently under investigation in two global phase III trials, namely the SILCAAT and the ESPRIT studies. PMID- 11059299 TI - [Hearing ability of Danish school children when starting and leaving school]. AB - School health records for a group of Danish school children, who started school in 1977, 1987 and 1997, were studied in order to evaluate hearing ability at the time of starting school and leaving school. One thousand, six hundred and five children who were evaluated by audiometry were included in the study. We found a higher prevalence of hearing loss in children who started school in 1987 and 1997 compared to those who started in 1977. The typical hearing loss was in the high frequencies. At the end of school the hearing ability of children who had started school in 1977 was just as poor as of that of those who had started in 1987. Whether this hearing loss may lead to poorer learning capacities in the afflicted children should be evaluated by further studies that include information on exposure to noise. PMID- 11059300 TI - [Active and passive smoking among personnel at the Bispebjerg Hospital 1992 1999]. AB - The purpose of this study was to describe changes in smoking behaviour and exposure to passive smoking among hospital employees at a large Danish University Hospital (Bispebjerg Hospital) from 1992 until 1999 as part of a program toward a smoke-free hospital. The study was based upon three cross-sectional self administered questionnaires surveys carried out among all employees at the hospital--approximately 4000 persons--in October 1992, April 1997 and April 1999, participation rates being 84, 80 and 76 percent. During the seven year period the smoking rate has decreased from 46% to 32% among male and 40% to 33% among female employees. A decrease in smoking rate was found among all subgroups of employees. Among male employees the rate of heavy smokers has decreased from 25 to 16%, among female employees this decrease is lacking, the rate of heavy smokers being 15% during the whole period. The numbers of employees exposed to passive smoking all day or most of the day has changed from 39% to 25% from 1992 until 1999. Among the smokers 30%--8% of all employees--responded that they would not be able to manage without smoking tobacco during working hours. This answer is most commonly found among heavy smokers, smokers with short or no education and smokers who smoke at any time of day. It is concluded that even though there has been a reduction in the smoking rate, the exposure to passive smoking among employees at the hospital still is unacceptably high. Based upon these results it has been decided that Bispebjerg Hospital is smoke-free for all employees from the 1st of January 2000. There is a need for initiatives for the smokers, who can't manage work without smoking. PMID- 11059301 TI - [Desmoid tumor in familial adenomatous polyposis]. AB - INTRODUCTION: Desmoid tumors (DT) are rare benign tumors that do not metastasize, but tend to invade locally. DT are frequently seen in patients with familial adenomatous polyposis (FAP), and diagnosis and treatment are often difficult. METHOD: The article presents the clinical picture, diagnosis and treatment of DT in patients registered in the Danish Polyposis Register by the end of 1999. RESULTS: Twenty-seven of 486 patients (6%) had DT. Eighteen patients were alive at the time of evaluation. DT were found in the mesentery in 42%, in the abdominal wall in 40%, in the retroperitoneum in 8% and only 10% on the extremities. Fifty percent of the patients had complications (intestinal obstruction, hydronephrosis or fistulas), and 2/9 deaths were caused by DT. Ninety-three percent were treated with surgery, NSAIDs, antioestogenic drugs, chemotherapy or radiotherapy, but all modalities proved disappointing, except for treatment with a combination of the NSAID sulindac and tamoxifen. Five patients treated with this combination showed extensive and long lasting response. DISCUSSION: Surgical excision is recommended in patients with DT in the abdominal wall. First line treatment of mesenteric DT is Clinoril in combination with tamoxifen. Elective surgery may be considered in patients with a small well defined DT with no signs of invasion of vital structures, and in patients with imminent bowel ischaemia or obstruction. The prognosis for mesenteric DT is grave, and improvement of the therapeutic strategy awaits current international studies. PMID- 11059302 TI - [Psychogenic paralysis. A prospective study]. AB - INTRODUCTION: Patients with motor conversion disorder are frequently seen in neurological departments. Long term prognosis is usually considered to be good, although earlier research has been somewhat unsystematic and mostly retrospective. This study follows a well investigated sample of patients for two to five years and attempts to identify predictors associated with prognosis. MATERIAL AND METHODS: Thirty patients with a recent onset of motor conversion disorder were assessed for key psychiatric and demographic variables. They were reassessed two to five years later. RESULTS: Nineteen patients had recovered completely and eight patients had improved, while only three patients were unchanged or worse. Contrary to other follow-up studies none of the patients received a rediagnosis of neurological disease. The presence of a personality disorder, concomitant somatic disease, and low DSM-IV axis V score proved to be associated with poor outcome. DISCUSSION: The results of this study stresses the need for careful and well-conducted neurological and psychiatric assessments in patients with psychogenic paralyses, bearing in mind the substantial possibility for coinciding illnesses. If this is ensured, it appears that the risk of subsequent neurological rediagnosis is negligible. PMID- 11059303 TI - [Good prognosis. Assessment of the course in patients with acute low back problems in the municipality of Copenhagen using medical audit]. PMID- 11059304 TI - [Epidural abscess after epidural catheterization. Frequency and case reports]. AB - Spinal epidural abscess is considered a rare but serious complication after epidural analgesia. In a recently published prospective study the incidence was found to be one in 1,930 catheters. Two case reports are presented and the high incidence of epidural abscess is discussed. PMID- 11059305 TI - [Artificial respiration in the treatment of heart arrest--is it necessary?]. PMID- 11059306 TI - [The season of ticks]. PMID- 11059307 TI - [Treatment of upper respiratory tract infections]. PMID- 11059308 TI - [Elimination or eradication?]. PMID- 11059309 TI - [Monoclonal interleukin-2 receptor antibodies and their use in clinical kidney transplantation: basiliximab and daclizumab]. AB - Basiliximab and daclizumab are analogous immunosuppressive agents approved for clinical renal transplantation. Both drugs are gene manipulated chimeric human/murine monoclonal antibodies with specificity against the alpha-chain of the interleukin-2 receptor (IL-2R) on activated T lymphocytes. By combining the drugs with calcineurin inhibitor based immunosuppression, an inhibition of the interleukin-2 (IL-2) synthesis as well as the IL-2/IL-2R reception is obtained. Consequently, the allogeneic immune response is suppressed and the risk of acute rejection of the transplant is reduced. By treatment with basiliximab or daclizumab in the recommended doses, a blockade of the IL-2R is obtained for up to 12 weeks. The incidence of acute rejection episodes is reduced by approx. 33%. The incorporation of human amino acid sequences has almost eliminated the immunogenicity of the drugs and reduced their rates of elimination. The side effects of prophylactic treatment with basiliximab/daclizumab do not differ from those of placebo. PMID- 11059310 TI - [Berries which burn]. PMID- 11059311 TI - [Long QT and torsades de pointes as adverse effects of drugs]. PMID- 11059312 TI - An enduring partnership. AJN and Lippincott enter: another century together. PMID- 11059313 TI - From the publisher. From 'invalid chairs' to online education, the journal has reflected the times. PMID- 11059314 TI - Letters excerpts of AJN classics. PMID- 11059315 TI - The third decade of the HIV epidemic. Highlights from the 13th International AIDS Conference. PMID- 11059316 TI - How can you bear to be a nurse? An AJN classic April 1987. PMID- 11059317 TI - 100 years in pictures. Images from 100 years of American nursing: celebrating a century of caring. PMID- 11059318 TI - 'I am faithful, I do not give out'. Walt Whitman, Civil War poet--and nurse. PMID- 11059319 TI - The campaign to eliminate the midwife. PMID- 11059320 TI - Humanizing childbirth. PMID- 11059321 TI - A nurse on Mars? Why not? PMID- 11059322 TI - Martha Rogers's odyssey. PMID- 11059323 TI - The TB preventorium. PMID- 11059324 TI - Emerging infectious diseases. PMID- 11059325 TI - Think different. Inventions and innovations by nurses, 1850 to 1950. PMID- 11059326 TI - 'The patient is awake'. PMID- 11059327 TI - Thermometers & telephones. A century of nursing and technology. PMID- 11059328 TI - The gene genies. PMID- 11059329 TI - Nurses making a difference. Two nursing pioneers. PMID- 11059331 TI - When care is needed at home. Your rights under the Family and Medical Leave Act. PMID- 11059330 TI - The wisdom of our practice. PMID- 11059332 TI - A global connection. The International Council of Nurses works to unify the profession. PMID- 11059333 TI - What we may expect from ethics and the law. PMID- 11059334 TI - Breaking the mold. The many legacies of nurses in progressive movements. PMID- 11059335 TI - Work for nurses in play-schools. AJN, October 1900. PMID- 11059336 TI - Don't get stuck with unsafe needles. Instead, get involved in needle device selection. PMID- 11059337 TI - Nursing & philately. PMID- 11059338 TI - The clinical utility of measuring total PSA, PSA density, gamma-seminoprotein and gamma-seminoprotein/total PSA in prostate cancer prediction. AB - BACKGROUND: To evaluate whether serum total prostate-specific antigen (PSA), PSA density (serum total PSA level divided by prostate volume), gamma-seminoprotein and gamma-seminoprotein/total PSA ratio could predict prostate cancer (PCa) prior to biopsy. METHODS: A total of 316 consecutive patients who had undergone transrectal prostate biopsy and/or transurethral resection were examined. The prostate volume was determined by transrectal ultrasonography (TRUS) and the ability of the above-mentioned four variables to distinguish PCa from benign prostatic hyperplasia (BPH) was evaluated. RESULTS: PCa was detected in 61 cases. Receiver-operating characteristic (ROC) analysis revealed that both the PSA density and serum total PSA were the most useful predictors of PCa among the four variables. For the patients with a serum total PSA level of 4.1-10.0 ng/ml, PSA density was significantly more accurate than total PSA (p < 0.005). An optimum PSA density value of 0.18 was chosen as a cutoff because it showed the highest sum of sensitivity and specificity, 92 and 54%, respectively. Using this PSA density cutoff, the number of biopsies could have been reduced to 57 from 63% when compared with a PSA density of 0.15. CONCLUSIONS: PSA density was significantly more accurate than other variables in predicting PCa. To avoid unnecessary biopsies, the PSA density cutoff value of 0.18 would be recommendable for determining a prostate biopsy for Japanese males with a serum total PSA level of 4.1-10.0 ng/ml. PMID- 11059339 TI - BRCA1 mutations in Taiwanese with epithelial ovarian carcinoma and sporadic primary serous peritoneal carcinoma. AB - BACKGROUND: Germline BRCA1 mutations of sporadic ovarian cancers are presumed to be rare events, except among specific populations. To date, the status of germline BRCA1 mutations in Taiwanese with primary epithelial ovarian carcinoma (PEOC) is still unknown. In this study, we tried to answer part of this question. METHODS: Sixty-four patients documented with PEOC, four patients with family history of breast and/or ovary cancer syndrome and five patients with sporadic primary serous peritoneal carcinoma (PSPC) were enrolled in this retrospective study from January 1994 through June 1999. At the same time, 50 normal healthy Taiwanese without family history were enrolled in this study. Germline DNA from these patients was screened for mutations in the BRCA1 gene using polymerase chain reaction-based single-stranded conformation polymorphism analysis (PCR SSCP). Shifting DNA bands were sequenced. RESULTS: One of the 64 patients with PEOC (1.6%) exhibited germline BRCA1 heterozygous mutation which was exon11 single-base substitution at nucleotide1047 (CAG to TAG). One of the five patients with PSPC (20%) exhibited an exon11 single-base substitution at nucleotide 914 (TCT to TCC) with resultant silent mutation. One of the normal healthy Taiwanese (2%) was found to have an exon 2 single-base substitution at nucleotide 152 (A- >C) which was also a silent mutation. No mutations of BRCA1 were detected in four patients with a family history of breast and/or ovarian cancer. CONCLUSIONS: Based on this study, it was very difficult to obtain precise data to prove the value of applying genetic testing of BRCA1 mutations in Taiwanese patients with sporadic epithelial ovarian cancers or sporadic PSPC and even with a family history of breast and/or ovarian cancer because of its rare event and because of the too small number of cases available in this study. PMID- 11059340 TI - Low-grade B-cell lymphoma of mucosa-associated lymphoid tissue in the thymus of a patient with pulmonary amyloid nodules. AB - Low-grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT-type lymphoma) is a rare thymic tumor, with only seven previous cases described worldwide to date. We describe the only case to have presented with pulmonary amyloid nodules. A 63-year-old Japanese female was found to have an anterior mediastinal tumor and multiple bilateral pulmonary nodules during a medical check up in 1990 followed by chest radiography and computerized tomography. Because the mediastinal tumor grew larger, she was referred to the National Cancer Center Hospital East and hyperglobulinemia was pointed out. The thymus was resected through median sternotomy and pulmonary nodules were also resected through left thoracotomy. The solid and nodular tumor with several small satellite extensions and cyst formation was completely confined to within the thymus and the resected pulmonary nodules consisted of solid masses with a rough surface. Histologically, monotonous medium-sized centrocyte-like cells occupied the medulla of the thymus and infiltrated Hassall's corpuscles (lymphoepithelial lesions) and the resected pulmonary nodules consisted of eosinophilic amorphous deposits which showed birefringence on Congo Red staining. Immunohistochemically, the tumor cells were positive for CD20 and CD79a. IgG and kappa light chain restrictions were also found in plasmacytoid cells in the tumor. Clonal rearrangement of the immunoglobulin heavy chain gene was demonstrated by polymerase chain reaction. We diagnosed this case as low-grade B-cell MALT-type lymphoma in the thymus and nodular pulmonary amyloidosis. Since the patient had only localized amyloid deposits in the lung far from the thymic malignant lymphoma and had high serum immunoglobulins, the pulmonary amyloid deposits might be derived from a circulating precursor associated with hyperglobulinemia. PMID- 11059341 TI - Hemangioma of the rib: a case report. AB - A case of hemangioma of the left seventh rib is presented. In January 1999, a 59 year-old woman presented with an enlarged costal mass which had been followed up for 4 years. Preoperative examination suggested chondrosarcoma because of tumor growth beyond the disrupted bony cortex. She underwent resection of the left seventh rib along with the sixth and seventh intercostal muscles and reconstruction of the chest wall defect. The pathological diagnosis of the lesion was hemangioma. She was discharged after an uneventful postoperative course. There has been no evidence of recurrence after a 14-month follow-up. Tumor growth beyond the disrupted bony cortex was a characteristic feature by both imagery and pathological examination in this case. This case represents a difficulty of a preoperative definite diagnosis of the chest wall tumors by imagery alone. PMID- 11059342 TI - Lung carcinoma with polypoid growth in the main pulmonary artery: report of two cases. AB - Invasion into the lumen of the main pulmonary artery is an uncommon mode of extension in lung carcinoma and its prognostic significance remains unclear. We describe here two resected cases of lung carcinoma that showed such a rare tumor spread. Although a preoperative evaluation, such as angiography or perfusion scan of the lung, had shown a significant decrease in circulation, we could not diagnose the intraluminal tumor growth preoperatively. Pneumonectomy was finally needed to perform a curative operation. The tumors were centrally located and showed polypoid growth in the main pulmonary artery. Postoperative pathological examination revealed the tumors to be adenosquamous carcinoma of the lung in both cases. No intrapulmonary metastases were detected. One patient is doing well with no signs of recurrence after a follow-up period of 10 years. Although intra arterial polypoid growth of lung carcinoma is extremely rare, such tumor extension should be considered preoperatively to perform a curative surgical resection, especially when the tumor is centrally located. While arterial invasion is generally an ominous prognostic factor, curative surgical resection would offer a good prognosis, even for lung carcinoma invading the main pulmonary arterial trunk. PMID- 11059343 TI - Chronic neutrophilic leukemia with acute myeloblastic transformation. AB - We report a rare case of chronic neutrophilic leukemia (CNL) which terminated in acute myeloblastic transformation 3 years after the onset of the disease. The increased leukocytes were mainly neutrophils at various maturational stages until 1 month before transformation without dysplastic hematopoietic cells or other myeloproliferative disorders. Repeated analyses for the Philadelphia chromosome (Ph1), rearrangement of the BCR gene or chimeric BCR/ABL mRNA, major, minor and mu, were negative. Genomic analysis of granulocyte colony-stimulating factor (G CSF) receptor did not reveal any abnormality. The clinical manifestations were characterized by hyperleukocyte syndrome with respiratory distress and ischemic legs with gangrene. PMID- 11059344 TI - Lessons in treatment of gastric cancer: viewpoint of a UK surgeon. PMID- 11059345 TI - Robotic surgery. PMID- 11059346 TI - Normal formation and development defects of the human dentition. AB - Oral health and systemic health are intimately related, and a thorough evaluation of the oral health of children is critical in providing appropriate health care. By understanding the normal sequence and patterns of tooth development, clinicians can readily identify children who deviate from normal dental development and provide appropriate interventions or make appropriate referrals. Developmental defects of the human dentition are not uncommon and can severely adversely affect the physical and psychological health of children. Despite the severity of some developmental defects of the dentition, the ability to diagnose and manage these conditions, in most cases, allows children the benefit of optimal oral health. PMID- 11059347 TI - Dental caries. An infectious and transmissible disease. AB - Of the infectious diseases that affect humans, dental caries may be the most prevalent. Pediatric primary medical care providers are usually the first health care providers to examine the oral cavity of a child and so need to be able to recognize suspicious dental lesions. This article provides information on recognizing caries and reviews the epidemiology, pathogenesis, and treatment of caries and odontogenic infections. PMID- 11059348 TI - Prevention of dental disease. The role of the pediatrician. AB - Evidence increasingly suggests that to be successful in preventing dental disease, we must begin preventive interventions within the first year of life. Pediatricians are well positioned to begin this process with early assessment of oral health and provision of anticipatory guidance, including ensuring that patients establish a dental home in addition to their medical home. This article provides information that will enable pediatricians to assess caries risk and provide practical and effective advice to parents about preventing dental disease, including oral hygiene, diet, and fluoride recommendations. PMID- 11059349 TI - Infant oral health and oral habits. AB - Many oral diseases and conditions, including dental caries (cavities) and malocclusions, have their origins early in life. Prudent anticipatory guidance by the medical and dental professions can help prevent many of the more common oral health problems. This article provides information on the rationale for early dental examination and instructions for pediatric and family practitioners in scheduling and conducting an early oral intervention appointment. In addition, feeding practices, non-nutritive sucking, mouth breathing, and bruxing are discussed, including their effects on orofacial growth and development. PMID- 11059350 TI - Diagnosis and management of dental injuries in children. AB - Approximately half of children sustain some type of dental injury. Management of injuries to the anterior teeth of preschool children is directed toward minimizing potential damage to the developing permanent teeth; therefore, heroic measures to save primary teeth are not indicated. Crown fractures in the permanent dentition, even those exposing the dental pulp, can be successfully treated hours after an injury. Prompt referral for dental treatment is advisable. Displacement injuries to permanent teeth constitute genuine dental emergencies in which the prognosis is directly related to the timeliness of treatment. Avulsed permanent teeth should be immediately reimplanted by any capable person. If that is impossible, the teeth should be placed in cold milk and the child referred for immediate treatment by a dentist. Mouthguards prevent dental injuries but are not widely used outside of a few organized sports. Efforts should continue to promote mouthguard use in all contact sports. PMID- 11059351 TI - Facial growth and management of orthodontic problems. AB - This article enables pediatricians to identify and understand the implications of common facial growth problems in children and adolescents. Problems with facial growth can result in aesthetic and functional concerns. Using a simple method of clinical evaluation, pediatricians can identify facial growth problems in the anteroposterior, vertical, and transverse dimensions. These problems can then be referred for evaluation and treatment by various means. Because facial growth is the result of the interaction of genetic and environmental factors (some of which are functional), growth modification may be a possibility. Some problems may be camouflaged or treated by combined surgical and orthodontic means. Continued growth in early adulthood may enhance or detract from treatment results obtained in childhood or adolescence. These dynamic properties of the face make management of facial growth challenging but generally rewarding and successful because of substantial aesthetic and functional improvements. PMID- 11059352 TI - Pediatric oral pathology. Soft tissue and periodontal conditions. AB - Pediatric patients can present with various intraoral lesions that require accurate diagnosis, treatment or reassurance, and possible referral for a dental evaluation. Periodic review of oral soft-tissue pathology can help the medical team to easily recognize common and rare abnormalities affecting children. Recent years have brought new insights into the causes and treatment of periodontal diseases of children, making prevention or treatment of many formerly untreatable conditions possible. Early detection of these oral conditions may be life saving. PMID- 11059353 TI - Relationships between oral and systemic health. AB - Oral and systemic health cannot be separated. This article addresses some established and emerging relationships that highlight the association between systemic and oral health. The physician's role requires an understanding of the effects of disease and its treatment on oral health. Also, physicians should be able to identify abnormality in the oral cavity that might be attributable to disease or be a compromising factor in the health, growth and development, or functioning of children and make a referral. Cooperation between dentist and physician can mean good overall health for children, including oral health. PMID- 11059354 TI - Pharmacologic behavior management for pediatric dental treatment. AB - Sedatives are an important and necessary management technique for some children during dental procedures. Sedation can be administered safely and efficiently by competent practitioners who have special training in the use of the technique in children and who adhere to sedation guidelines. Nonetheless, some children present with special needs, such as medically compromising conditions, or multiple carious teeth in a child who is fearful or whose family must travel a long distance for care. Sedation cannot always safely and adequately meet the needs of these children. In these cases, general anesthesia in a hospital, ambulatory care facility, or office is indicated and must be provided by an appropriately licensed and trained physician or dentist. PMID- 11059355 TI - Public and clinical policy considerations in maximizing children's oral health. AB - Although children's oral health in the US has improved over recent decades, a subset of children continues to suffer extensive disease that is severe enough to constitute a public health problem with considerable individual consequences. Disparities in oral health are wide and growing. Dental caries prevalence may be increasing with shifts in child demographics. Policy approaches to maximizing children's oral health and pediatricians' roles in promoting effective policies are discussed. PMID- 11059356 TI - Does the GMC fulfil its mission statement--"protecting patients, guiding doctors"? PMID- 11059357 TI - Do NICE and CHI have no interest in safety? Opinion of the book NICE, CHI and the NHS reforms. Enabling excellence or imposing control? AB - Seventeen eminent and experienced people have contributed to this most valuable review of NICE and CHI and their potential impact on clinical practice in the UK. There is essentially 100% agreement that the basic concept is a good one; we all want to have the highest possible quality of clinical practice and improvements in health care. This is all motherhood and apple-pie stuff which goes without question but the problem is how it is put into effect. The contributors are also in agreement and fear that central desire for control will outweigh the benefits. The most recent NICE action, which was leaked to the media as a 'preliminary opinion', concerned the use of beta-interferon for the treatment of multiple sclerosis (MS). The opinion seems to be that beta-interferon is very expensive, that, yes, it does help some sufferers but, no, it does not help others and because it costs more than the NHS can afford no one can have it. This seems to me to be a most unsatisfactory outcome. Surely what clinical excellence demands is the refinement of diagnostic capabilities so that those who will benefit may be distinguished from those who will not. In the meantime we do the best we can even if it does mean that the NHS has to pay for some patients who do not respond. This is the inevitable consequence of the belief that a 'free' and comprehensive health service can be provided out of general taxation. Beta interferon for the treatment of MS is an example of the observable fact that medical science is advancing at a rate considerably in excess of possible increases in funding. Possibly the most important problem identified in this book is the absence of a relevant, high quality data source for the preparation of the numerous guidelines that NICE is expected to produce each year. In a fully grown science a starting point for a quantitative procedure is the establishment of a baseline and, having done that, the scientist's next step is to produce a standard curve for use in the measurement of further investigations. I have said previously that medicine is not a fully grown science (which is one of the problems) but that does not mean that basic scientific method can be abandoned. What is the baseline for the evaluation of clinical practice? The best would be records of the progression of a disease-state in untreated patients. That, for obvious reasons, is clearly not possible so a compromise is unavoidable. Unfortunately we do not even have that compromise baseline so how do we know what is better and what is worse? In simple, single disease states and within the limits of RCTs that is sometimes possible but in a population composed of many elderly people with multiple pathology it is greatly more difficult. If NICE is to produce authoritative guidelines then its first task is to define a (compromise) baseline. For the readers of this journal the absence of safety as one of the measures of clinical excellence must be a matter of concern. All clinical interventions may be casually related to adverse reactions which may, on occasion, be serious or even fatal. Perhaps excluding safety was a conscious decision by those who created NICE. At the time of market approval information on safety is almost always limited to events occurring more frequently than 1 in 1000 exposures which is far below the desirable level of precision. If NICE is to provide advice at the time of market authorization or shortly after then it will never be in a position to include an acceptable evaluation of safety. So why give a hostage to fortune by mentioning it in NICE's remit? Time alone will tell whether NICE and CHI achieve health improvement or whether they prove to be no more than a political gesture. PMID- 11059358 TI - Prepulsid withdrawn from UK & US markets. PMID- 11059359 TI - Drug regulation in the European Union. PMID- 11059360 TI - Resistance to antimicrobial agents: can we make a difference? PMID- 11059361 TI - Therapeutic applications of medicinal plants in the treatment of breast cancer: a review of their pharmacology, efficacy and tolerability. AB - Various active compounds (or their semi-synthetic derivatives) derived from medicinal plants have been assessed for their efficacy and tolerability in the treatment of breast cancer. Some of these plant species, including Taxus baccata (paclitaxel, docetaxel), Podophyllum peltatum (etoposide), Camptotheca acuminata (camptothecin) and Vinca rosea (vinblastine, vinorelbine) have well recognized antitumour activity in breast cancer, and have been evaluated in clinical trials. For example, results from recent Phase II/III trials have established docetaxel as the most active single agent in the treatment (first or second-line) of advanced metastatic breast cancer. For other plant species such as Panax ginseng and Allium sativum, antitumour activity has been evaluated in experimental studies using cultured cells and animal models, but the therapeutic potential in patients remains to be determined. Antitumour activity derived from medicinal plants may produce results via a number of mechanisms, including effects on cytoskeletal proteins which play a key role in mitosis (paclitaxel), inhibition of activity of topoisomerase enzymes I (camptothecin) or II (etoposide), stimulation of the immune system (Viscum album), or antiprotease-antioxidant activity. Medicinal plant-derived antineoplastic agents may be used in single agent or in combinational therapies, and have been used in first-line or second line (including anthracycline-refractory patients) treatment of localized or metastatic breast cancer. Adverse effects resulting from the use of these agents include neutropenia and peripheral neuropathies. PMID- 11059362 TI - Modern management of acute myocardial infarction. PMID- 11059363 TI - Cosmetics and skin care products. A historical perspective. AB - The history of cosmetics and skin care products parallels many important technologic developments in chemistry, materials, and packaging innovations. Much is revealed about ancient civilizations by the cosmetics that are uncovered in archaeologic excavations. Much can also be said about modern-day health and adornment practices based on products in the current marketplace. PMID- 11059364 TI - Modern skin cleansers. AB - The course of development of skin cleansers has been one of continual improvement. Soap-based products, used since antiquity, offered improved cleansing over mechanical methods or water alone but could irritate and dry skin. Bars based on synthetic detergents that offer improved skin compatibility compared with soap have become available over the past several decades. Body washes have been growing in consumer popularity. Some of the first body washes introduced into the market offered a moisturization benefit in addition to mildness. Some second-generation body washes that are now on the market use even more sophisticated formulation schemes, such as coacervate technology, to deliver emulsified petrolatum to the skin during washing, providing mild cleansing and a significant dry skin improvement benefit. Consumer demand and the formulation possibilities provided by new product formats, new technologies, and new ingredients will undoubtedly lead to the delivery of even greater skin benefits in the future. PMID- 11059365 TI - Modern approaches to photoprotection. AB - UV light reacts with skin to produce undesirable changes, including photoaging and skin cancer. Sunscreen strategies are useful for protection against UV-B and short-wave UV-A, but complete protection against long-wave UV-A has not been achieved. Because UV-A is especially efficient at generating reactive oxygen species, it is being recognized increasingly as an important cause of photoaging and skin cancer. PMID- 11059366 TI - Tanning preparations. AB - Increasing consumer awareness as to the hazards of UV light should fuel ongoing interest in self-tanning products. As a result of the benign toxicologic profile of DHA, products containing DHA represent a safe alternative to UV-induced tanning. The results obtained with these products depend on the final formulation, the individual application technique, and the consumer's complexion type. Greater experience in formulation combined with increasing sophistication on the part of the consumer should lead to continuing growth in interest and satisfaction with the use of DHA-containing sunless or self-tanning products. Individuals need to be informed that these products do not offer significant protection against UV-B. If formulated with standard sunscreens, consumers should be cautioned that the duration of UV protection is more short-lived than the color change. PMID- 11059367 TI - Therapeutic moisturizers. AB - Moisturizers have been adapted to perform many important roles on the skin surface. Simple moisturizers combine occlusives and humectants to enhance the water-holding capacity of the skin. The addition of carefully selected emollients can influence the esthetic properties of the moisturizer and the stability of the active ingredients. The addition of sunscreens to moisturizers has created a new product category with an added skin function. Further diversity in moisturizer formulation is created through the addition of specialty ingredients, designed to enhance the functioning of the skin. Moisturizers are an important part of the dermatologist's armamentarium. PMID- 11059368 TI - Cosmeceuticals. AB - The author uses kinetin, a plant-derived nucleotide, as an example to summarize the approach to advising a patient on a new product. (1) Does it penetrate the stratum corneum? Topically applied nucleotides can penetrate human skin, and one of the most active and useful of these for the treatment of actinic keratoses, 5 fluorouracil (5-FU), is used widely in dermatology. 5-FU derives some of its benefit from its uptake, specifically in actinic keratoses, which do not have a complete epidermal barrier. The molecule is able to penetrate the stratum corneum. It is not clear that topically applied nucleotides have the same degree of penetration on photodamaged skin without actinic keratoses. (2) Is there a plausible biochemical mechanism of action? 5-FU has a well-known mechanism of action (i.e., the inhibition of DNA/RNA synthesis by incorporation of a false pyrimidine analog). The mechanism of action for furfuryladenine in human skin remains unknown and needs to be shown. If furfuryladenine functions as an antioxidant, it may be useful as a photoprotectant. This function does not account for a mechanism of action for the reversal of photoaging, however. (3) Are there published peer-reviewed, double-blinded, placebo-controlled, statistically significant clinical trials to substantiate the efficacy claim? Peer-reviewed, double-blinded, statistically significant clinical trials on furfuryladenine have not been published to date. The field of cosmeceuticals presents a quandary. The list of cosmeceuticals for the dermatologist to assess grows longer each year. It is possible that some of the active ingredients are beneficial physiologically in human skin and that they can offer specific benefits, such as photoprotection. Further research needs to be conducted to validate these claims. It also is likely that a wide variety of molecules purported to be active in human skin do not have any physiologic benefit in human skin. Only further research can answer these difficult questions. PMID- 11059369 TI - Sebum, cosmetics, and skin care. AB - Recent contributions to the understanding of the physiology and biochemistry of sebum production and of the lipids on the skin surface are leading to the development of effective strategies to regulate excessive sebum production and its consequences through the use of cosmetics and skin care products. PMID- 11059370 TI - Colored facial cosmetics. AB - Colored cosmetics are an important part of the dermatologic armamentarium. They can camouflage contour and pigment abnormalities, provide moisturization, enhance oil control, add sun protection, deliver barrier-enhancing agents, increase acne treatment, and create a sense of personal well-being. Familiarity with these products allows the dermatologist to provide better patient care. PMID- 11059371 TI - Eye cosmetics. AB - There are many eye cosmetics available to enhance the beauty or improve the appearance of the face. To prevent infection, most eye cosmetics contain preservatives. Fragrance is usually absent to keep the products as safe as possible. Hypoallergenic products contain fewer ingredients and may be more appropriate for patients with sensitive skin. PMID- 11059372 TI - Lip cosmetics. AB - Throughout the centuries, the beauty of lips has been extolled by poets, and painters; and people in many different cultures decorate their lips. Modern cosmetics are designed not only to beautify the lips but also to moisturize and protect them from environmental hazards. Familiarity with the ingredients used in lip cosmetics is essential to recognizing and diagnosing the adverse reactions that are associated with these products. PMID- 11059373 TI - The biology of hair care. AB - Most hair damage occurs as a result of grooming habits and chemical exposure for cosmetic purposes. An evaluation of hair structure and biology points to the need for better protective mechanisms from cuticular damage and UV damage to maintain the cosmetic value of the hair. Through the cooperative efforts of dermatologists and cosmetic chemists, better hair care products can be developed. The dermatologist needs to elucidate the mechanism through which products can enhance hair shaft functioning, whereas the cosmetic chemist needs to identify substances and develop formulations to accomplish the desired end. PMID- 11059374 TI - Skin and hair cosmetic issues in women of color. AB - Women of color comprise many phenotypically heterogeneous groups. Despite the general heterogeneity, however, there are unique skin and hair care issues and needs. These issues often present therapeutically challenging problems for the dermatologist and the skin and hair care industry. PMID- 11059375 TI - Appearance, cosmetics, and body art in adolescents. AB - Appearance in adolescents is a means of communication, a language expressing self identity. Teenagers explore fashions to make personal statements. Teenagers are significant consumers of various toiletry and skin care products that fill their cleansing, hydrating, and photoprotective needs. They also are enthusiastic consumers of products aimed at adolescent fads, such as decorative hair and nail cosmetics. For some teenagers, the expression of individualism is achieved through body art, such as tattooing and body piercing. Areas of concern are the lack of motivation for sun protection and the risky behavior associated with body piercing and tattooing. PMID- 11059376 TI - Nail cosmetic issues. AB - Nail cosmetics are a part of good hygiene and self adornment. These cosmetics can provide solutions to problems involving the nail plate and also can create disease. This article provides a diagnosis and treatment-oriented approach to nail cosmetic issues. PMID- 11059377 TI - Adverse reactions to cosmetic ingredients. AB - Numerous cosmetics are used on a daily basis by men, women, and children. Despite a few adverse reactions, cosmetics are a remarkably safe group of products. PMID- 11059378 TI - Cosmetics. A dermatologist looks to the future: promises and problems. AB - The skin-care industry has created myriad new products to meet the needs of an aging population. Novel "bioactive" ingredients are derived from the sea, the earth, and the plant kingdom. Popular ingredients include Chinese herbs, vitamins, minerals, antioxidants, enzymes, hormones, and a multitude of "naturals". For academic medicine, this avalanche of new products poses the task of establishing international safety standards for determining how much product efficacy is science-based and how much is marketing hype. PMID- 11059379 TI - The use of psychotropic drugs in dermatology. AB - In approximately one third of dermatology patients, the effective management of a skin condition also involves consideration of associated emotional and psychosocial factors. Psychotropic drugs are an important part of the dermatologist's therapeutic armamentarium. This article reviews some of the salient pharmacologic guidelines and important drug-drug interactions involved in the use of the major classes of, for example, antidepressants and antipsychotic drugs, in the dermatologic patient. PMID- 11059380 TI - [Disorders of endothelium-dependent vascular reactions and of the arginase and NO synthase pathways of L-arginine metabolism in arterial hypertension]. AB - In normotensive rats (NTR) and spontaneously hypertensive rats (SHR) with high (subgroup 1) and low (subgroup 2) level of the systemic arterial pressure (SAP) we studied an activity of arginase and nitric oxide synthase (NOS) in different tissues, and the content of their metabolites: urea and nitrit anion (NO2-). In isolated preparations of a thoracic aorta we recorded endothelium-dependent (ED) dilator reactions on acetylcholine (Ach). It has been found that in heart, aorta, plasma and erythrocytes of rats (subgroup 2) both the activity of arginase and content of urea increase remarkably. In heart, the activity of arginase reaches 27.96 +/- 5.92 nmol.min-1.mg-1 of protein, in aorta 4.74 +/- 0.99 nmol.min-1.mg-1 of protein (as compared with NTR 1.32 +/- 0.12 nmol.min-1.mg-1 of protein and 1.12 +/- 0.07 nmol.min-1.mg-1 of protein, accordingly). Content of urea in heart reaches 679.5 +/- 121.19 nmol.mg-1 of protein, in aorta 350.6 +/- 63.6 nmol.mg-1 of protein (in NTR it was 36.8 +/- 5.3 nmol.mg-1 of protein and 43.02 +/- +/- 9.55 nmol.min-1.mg-1 of protein, accordingly). It was followed with a decrease in the NOS activity and heterogeneous changes in NO2- content in the tissues under exploration. For example, the activity of NOS in heart and aorta decreased to 0.018 +/- 0.005 nmol.min-1.mg-1 of protein, in aorta 0.183 +/- 0.037 nmol.min 1.mg-1 of protein, accordingly, as compared to 0.093 +/- 0.014 nmol.min-1.mg-1 of protein and 0.41 +/- 0.07 nmol.min-1.mg-1 of protein in NTR. Content of NO2- in aorta decreased by 0.79 +/- 0.06 nmol.mg-1 of protein, but in heart it increased to 0.63 +/- +/- 0.13 nmol.mg-1 of protein, (in NTR it was 2.15 +/- 0.18 nmol.mg-1 of protein and 0264 +/- 0.04 nmol.min-1.mg-1 of protein, accordingly). In rats, subgroup 2, ED dilator responses of the smooth muscle (SM) of the thoracic aorta were inhibited by Ach (10(-6) mol). Their amplitude reduced by almost twice, and a latency for their response became 4 times as much. All the changes in the biochemical parametres in heart, aorta, plasma and erythrocytes, and changes in contractile activity of vascular SM proved to be also characteristic for rats in subgroup 1, but they were less expressed quantitatively. Thus, for the first time we have studied an activity of two alternative pathways for the metabolism of L arginine on the model of arterial hypertension. The data obtained evidence that at hypertension non-oxidative (arginase) pathway of L-arginine metabolism is activated, while the oxidative pathway (NOS) is inhibited. Changes in the balance between them are followed with an essential inhibition of ED vasodilator responses. All this give us the prove to think of the origin for the arterial pressure increase to be both genetically and quantitatively determined damages in the biochemical homeostasis and dependent on it endothelial regulation of vascular tone. PMID- 11059381 TI - [Lipid peroxidation and the antioxidant system activity of the blood serum and tissues of the stomach and brain in interoceptive stimulation of the gastrointestinal tract]. AB - Changes of the level of lipids peroxide oxygenation and activity of antioxigenative system in blood serum and in the tissues of stomach, brain under the condition of interoceptive stimulation of the gastro-intestinal tract. Violation of correlation among the processes of free radical lipid oxidation and antioxidant defence of the organism towards activation of lipoperoxidation in the above mentioned conditions has been determined. The degree of this violation depends on the nature of the stimulus action. PMID- 11059382 TI - [The correlations between the morphometric indices of immunocompetent organs, the adrenal glands and the peripheral blood cells in different types of adaptive reactions in an experiment]. AB - The correlation coefficients between the masses of the thymus, morphometrics of the thymus, the masses of the spleen, of the adrenals and the lymphocytes, the eosinophils of the peripheral blood was studied in 92 white rats by the adaptation reactions: stress, orientation, activation, defective adaptation. The different changes depend on the type of the adaptation reaction. PMID- 11059383 TI - [The characteristics of the effect of thyroxine on sphingosine and phosphoinositide synthesis in the rat liver in ontogeny]. AB - The age peculiarities of sphingosine, phosphatidylinoside, phosphatidylinoside 4 phosphate and phosphatidylinoside 4,5-bisphosphate in liver and thyroxine influences on this processes has been investigated. It has been determined that sphingosine, phosphatidylinoside and phosphatidylinoside 4-phosphate synthesis in liver drop in old age. It was found that thyroxine enhances the labelled precursor incorporation into sphingosine of 3- and 24-old rat, and phosphatidylinoside 4,5-bisphosphate--24-old animals and decreases the synthesis of phosphatidylinoside and phosphatidylinoside 4-phosphate in liver of the young rats. PMID- 11059384 TI - [Sex steroids in the initiation of puberty genesis in female rats]. AB - In the female rats it has been estimated the initial (vaginal opening) and final (the establishment of the positive feedback and ovulation) stages of the sexual maturation are in most cases separated by time and regulated by different testosterone metabolites. The considerable decrease of testosterone and its biotransformation to 5 alpha-reduced androgens are necessary for induction of initial puberty stage. Duration between the beginning and the ending of the pubertogenesis depends on the way of the testosterone metabolism. The transformation of testosterone to 5 alpha-reduced androgen stimulates the vaginal opening, but enlarges the time of the first ovulation. The activation of testosterone aromatization promotes to the shortening of the time interval or even the temporal coincidence of the initial and final puberty stages. PMID- 11059385 TI - [The electrical activity of the hypothalamic arcuate nucleus and the morphofunctional characteristics of the gonads in birds during blockade and stimulation of the beta-adrenergic receptors]. AB - Beta-adrenoreceptors agonist isadrine as shown by measurement of hypothalamic arcuate nucleus (AN) summary electrical activity and by morphometry of its neurons and testis does not result in significant acceleration of maturation in maturing (4-6 week old) male japanese quails (Coturnix coturnix japonica). Anapryline-induced blockade of beta-adrenoreceptors leads to small inhibiting of the reproductive complex. This findings confirm our previous works, according to which brain beta-adrenergic system does not play leading role in regulation of maturation in birds. The direct correlation between morphometrical and electrophysiological features of AN neurosecretory cells was demonstrated as well. PMID- 11059386 TI - [The role of lysosomal proteinases in blood system reactions during inflammation]. AB - On the model of carrageenan-induced acute aseptic peritonitis in rats it is shown that in inflammation on the background of contrykal administration the accumulation and stimulation of leukocytes of an inflammatory focus, and the activation of granulomonocytopoiesis and of blood leukocytes are increased, and infiltration becomes calm earlier. The results indicate that in the natural development of inflammation lysosomal proteinases limit the accumulation and stimulation of leukocytes of an inflammatory focus, and activation of granulomonocytopoiesis and of blood leukocytes, and prolong the inflammatory reaction, i.e. they inhibit the protective adaptive reactions of blood in inflammation. PMID- 11059387 TI - [The effect of the preparation Wobe-Mugos E on the functional and biochemical status of the kidneys in the polyuric stage of sublimate-induced nephropathy]. AB - The protective effect of Wobe-Mugos appliance on the kidney function and biochemical state in polyuric stage of sublimate nephropathia at the moment of tubulointerstitial component formation was revealed in experiments on 40 white male rats. It appeared in the increase of hydrogenous ion excretion, titred acids, renal tissue fibrinolytic and proteolytic activity. The succinatdehydrogenase activation in renal cortex matter pointed out on the improvement of energy balance. PMID- 11059388 TI - [The effect of sodium alpha-ketoglutarate on the indices of the peripheral blood and lipid peroxidation and on the enzyme activity of antioxidant protection in irradiated rats]. AB - The influence of chronic roentgen irradiation in low doses on rats' quantitative and qualitative indices of peripheral blood, on lipid peroxidation and on erythrocyte antioxidant enzymes activity has been studied. It was shown that chronic roentgen irradiation in low doses had a destabilizing influence on leucocytes correlation, activated lipid peroxidation, depressed activity of erythrocytes antioxidant enzymes. Alpha-ketoglutarate injection in therapeutic doses normalized blood indices, limited the intensity of lipid peroxidation and activated antioxidant system enzymes. PMID- 11059389 TI - [The effect of magnetic-laser irradiation on the state of the cardiac muscle in adrenaline-induced myocardiodystrophy and food deprivation]. AB - In experiment on white rats was evaluated the positive effect of food deprivation on heart damaged by norephynephrine. Magneto-laser irradiation increased the adaptive resources of cardiovascular system during food deprivation and norephynephrine myocardiodystrophy. PMID- 11059390 TI - [Changes in the lysosomal apparatus of the neutrophilic leukocytes in the disseminated coagulation syndrome]. AB - The dependent of DIC development on lysosomal apparatus neutrophils activity was founded. The most severe changes in hemostasis system were registered in the period of maximum acid phosphatus activity. PMID- 11059391 TI - [The production of antihypertensive peptides in beta-casein proteolysis]. AB - Antihypertensive peptides were obtained after the proteolysis of beta-casein by starter cells Lactococcus lactis ssp. lactis and lactococci with pepsin or fromase. The peptides have shown the effect as inhibitors of angiotensin converting enzyme. The strongest action the peptide obtained after the proteolysis of beta-casein by synergic action of lactococci with pepsin has shown. It demonstrates a capability of formation of such peptides directly in milk products during their making and maturation under the action of proteolytic system of lactic acid bacteria and milk clothing proteases. PMID- 11059392 TI - [The effect of thyroxine on the enzymatic activity of the energy metabolism and antioxidant system in the neutrophilic granulocytes of piglets]. AB - The influence of thyroxine on activity of enzymes of energy metabolism (hexokinase, phosphofructokinase, pyruvate kinase, laktate dehydrogenase, glucose 6-phosphate dehydrogenase, NADP-isocitrate dehydrogenase, cytochrome-c oxidase) and antioxidative system (glutathione peroxidase, glutathione reductase, superoxide dismutase) of neonatal piglet neutrophils was investigated. It has been found, that after durable injections of hormone (4 mg/kg body weight) the increase of glycolytic enzymes activities as well as aerobic energy pathway catalyzers took place. Simultaneously the augmentation of superoxide dismutase reaction occurred after the thyroxine treatment. Such effect might represent an important link in compensatory mechanism, which prevents the destructive action of reactive oxygen species. PMID- 11059393 TI - On gratitude and gratification. AB - Expressions of gratitude from the patient may be regarded by the analyst as a much-needed validation or affirmation of competence. The analyst's need for gratitude may be a relatively silent presence when things are going smoothly or when the analyst's efforts are openly appreciated by the patient. Ungrateful patients, however, are likely to force the analyst to confront his or her unconscious background wish to enact a longed-for mode of relatedness as part of the daily work of psychoanalysis. The analyst's wish for a specific form of object relationship involving a selfless, devoted helper and an appreciative patient who acknowledges having been helped may be thwarted by certain patients at every turn, knowing they are depriving the analyst of a particular form of gratification in their work. For patients of this type, failure may mean success. The implications of this particular form of clinical stalemate are outlined, and a clinical example illustrates some of the challenges encountered in psychoanalytic work with ungrateful patients. PMID- 11059394 TI - Letting go: some thoughts about termination. AB - Termination of analysis is discussed from three perspectives. First, considered as a vicissitude of the analytic relationship, termination contains essential elements of the psychoanalytic process itself. Cycles of attachment, loss, mourning, and internalization mark moments in, as well as overviews of, every analysis from its beginning to well past its termination. Second, Freud's approach to the subject of termination is explored and widened, with an emphasis on its relation to mourning and on the depth and permanence of analytic transference--two dimensions relatively neglected by Freud, perhaps for personal reasons. Finally, clinical issues are presented that are meaningful to the author in his work with analysands, including his work as a training analyst. PMID- 11059395 TI - Suggestion and psychoanalytic technique. AB - The role of the analyst's suggestive influence on the course and outcome of psychoanalytic treatment is explored, and traditional and newer perspectives on analytic technique are contrasted. The intersubjective critique of the neutral, objective analyst in relation to suggestion is examined. The inevitable presence and need for suggestive factors in analysis, and the relationship of suggestion to transference susceptibility, are emphasized. The manner in which the analysis of suggestive factors is subsumed in transference analysis as part of traditional technique is highlighted. PMID- 11059396 TI - Future and potentiality in the psychoanalytic process. AB - A perspective is delineated on the dimension of the future in the psychoanalytic situation. Clinical manifestations are presented of the tension between actuality and potentiality that characterizes the treatment situation. This tension, an aspect of the intersubjective field that exists between patient and analyst, involves the analyst's hopes, expectations, anticipations, sense of purpose, and therapeutic intent, facets of the analyst's subjectivity that affect the clinical process. The question of the patient's individuality and autonomy is raised in the context of the notion of the "true self." To understand potentiality in the clinical situation, it is argued, the intersubjective emphasis on the inevitable mutual influence between analyst and patient must be complemented by a view of the self as emerging from within and gaining coherence through the unfolding of inherent dispositions and potentialities. PMID- 11059397 TI - The wish to regress in patient and analyst. AB - The analyst's wish to regress is used as a paradigm of the "forbidden" topic of what analysts want from their analysands. The aim is to expand the subjective domain of analysts' awareness so that they can analyze better by grasping more of their temptations with patients before enactment can occur. Clinical examples illustrate how the author temporarily joined patients in wish-fulfilling mutual regression. Analytic process is disrupted when the analyst wishes to relinquish the more differentiated role of the containing and interpreting analyst in favor of more childlike relatedness both with the patient and with the analyst's internal objects. The author, expecting a more typical counter-transference, had not anticipated that he might temporarily join these nonpsychotic patients in mutual regression. It is suggested that in the face of analytic impasse analysts should consider whether they might temporarily have joined the patient in mutually regressive wishes that have taken them away from more responsible analytic functioning. PMID- 11059398 TI - Passion, countertransference enactment, and breakdown in the psychoanalysis of a young woman. AB - Transference-countertransference involvement with patients the analyst finds "pleasurable" is examined. Analysis of such patients courts the danger that pathological identifications will go unrecognized and the analysis end at a point where the opportunity for decisive change, which to the patient appears physically and psychically catastrophic, is forgone. A presentation of one aspect of the analysis of a young woman reveals the psychic pain experienced by analysand and analyst when the feared breakdown places a pathological identification in doubt, and the difficulties that can ensue in treatment. Other psychodynamic lines of interpretation and differential diagnostic considerations are put aside in favor of an examination of the special role of enactment and countertransference enactment and their impact in promoting development or strengthening defense. PMID- 11059399 TI - The consultation process with an adult: a child analyst's perspective. AB - A close examination of a consultation is presented. Such a face-to-face encounter with an adult patient can suggest a parallel with another type of treatment: child analysis. It is argued that certain ways of working with children in analysis--the gliding between the interpersonal and the intrapsychic--were especially useful in bringing this adult patient to engage in analysis. Also highlighted in this account are interventions that function not as interpretations per se but as protointerpretations aimed at arousing preconscious affect states for the purpose of bringing them into conscious experience. PMID- 11059400 TI - The influence of the gender of patient and analyst in the psychoanalytic relationship. AB - An overview of current explorations of the influence of gender on the psychoanalytic situation is presented. The topic does not lend itself to simple generalizations because the accumulated experience of several generations of psychoanalysts is now merging with newer views of psychoanalytic technique, while at the same time psychoanalytic theory is being influenced by changing ideological crosscurrents and by new knowledge regarding the similarities and differences in the development of both genders. Major issues still open are the relationship between gender and sexuality and between erotic desire and love; the challenges of the boundary of the psychoanalytic relationship as a facilitating and containing frame for the exploration of oedipal conflicts; and the related temptations, prohibitions, and derivatives of the erotic tension in the transference-countertransference. PMID- 11059401 TI - Patient-therapist match: revelation or resistance? AB - Patient-therapist match is a relatively new yet frequently invoked concept within psychoanalysis. Despite Freud's appreciation of the influence of the analyst's past to his or her work within the analytic setting, psychoanalysts have historically held varied opinions about the degree to which the analyst's personality and conflicts affect the analytic process. As analysis was reconfigured as a two-person system, attention focused on the fit between patient and analyst. The literature on patient-therapist match is reviewed, and the conclusion reached that this intuitively appealing concept suffers from a lack of rigorous definition and operationalization. Many authors invoke match in ways that imply that it is real, static, external to the domain of analytic inquiry, and unaffected by analytic process. In its present form, the concept of patient therapist match obstructs rather than facilitates analytic exploration and obscures rather than clarifies what happens between analyst and analysand in psychoanalysis. By suggesting that match exists as a reality outside the domain of transference and countertransference, analysts may overlook the importance of psychoanalytic technique in creating a sense of match. Analysts may attribute stalemated or limited analyses to a bad match, rather than tenaciously exploring the transference-countertransference configurations that remain at the heart of analytic work. PMID- 11059402 TI - A return to basics: psychoanalytic conceptualization of motivation. PMID- 11059403 TI - The mechanism of action of psychoanalytic treatment. PMID- 11059404 TI - Modern defense analysis: three perspectives on the same hours. PMID- 11059405 TI - [Ability to work--new approach to evaluation methods]. AB - In Poland, the assessment of work ability has thus far been almost entirely objective, which means that it was based on the evaluation of the individual's health state. That is why a subjective method of work ability assessment with work ability index (WAI), developed by the Occupational Health Institute in Helsinki, was used in our present study. This method allows to indicate other possible factors which modify work ability. The study covered 189 men employed in five metallurgical plants, located in the region of Lodz. In the study population, WAI and work load on the basis of the expenditure of energy were measured, the health condition was evaluated, and information on life styles and non-occupational responsibilities was gathered. It was found that WAI values were inversely proportional to age and work load. They were also modified by individual characteristics, such as life style, body mass, and activities at leisure. It was found that the correlation between the general index of work ability and the objective health indicators was low with the concomitant considerably higher correlation with the values of the components which reflect subjective work abilities. In view of the results obtained, WAI can be recommended as a tool for assessing work ability. Due to this method it is possible to present conclusively all elements of individual characteristics and to identify at the same time links with working conditions. PMID- 11059406 TI - [Chemical hazards in fire-fighting environments]. AB - The assessment of the fire-fighters' exposure to harmful chemicals during the fire attendance is presented. The assessment was based on measurements of carbon monoxide, nitrogen dioxide, sulfur dioxide, formaldehyde and aromatic hydrocarbons of five fire-fighting actions. Using passive dosimeters, personal air samples were collected. A portable Draeger-PAC III was used for measuring carbon monoxide. Above 130 chemicals were detected in the environment of the fire attendance. Among them aliphatic hydrocarbons C6-C16 were dominant. Benzene and its aliphatic homologues were also found in all air samples. The carbon monoxide concentration accounted for up to 720 mg/m3. Concentrations of nitrogen dioxide, sulfur dioxide, formaldehyde and benzene ranged from 0.0 to 49.9 mg/m3; 84.5 mg/m3; 5.3 mg/m3 and 89.4 mg/m3, respectively. PMID- 11059407 TI - [Diagnostic neurologic procedures and certification in persons chronically exposed to carbon disulfide in light of personal cases]. AB - Carbon disulfide is a poison of particularly neotropic properties. In order to diagnose chronic occupational intoxication with carbon disulfide, a very careful examination of the central and peripheral nervous systems in required. The presence of subjective disorders only does not as yet provide grounds for diagnosing chronic intoxication. Organic changes like chronic encephalopathy or polyneuropathy after excluding the so called 'idiopathic' neurological diseases, may serve as a basis for certifying occupational disease. PMID- 11059408 TI - [Microflora of the farming work environment as an occupational risk factor]. AB - The paper presents the results of the studies concerning the air mould fungi concentration in the work environment during grain threshing, flax breaking and thyme cleaning. A high level of fungi was observed in the air at all workplaces with the highest concentrations during grain threshing. Of the total number of 12 farms, in 8 the air mould fungi concentration exceeded the proposed standard of 50,000 CFU/m3. Among the species containing cereals, flax and thyme, mould fungi of Alternaria alternata sp.--an etiologic factor of allergic respiratory diseases -occurred most frequently. During flax breaking, the presence of fungi of Fusarium poae sp., which affect grain husks and cause a great economic loss in agriculture, was observed. Mould fungi of Aspergillus candidus sp. were found during thyme cleaning. These fungi have been reported to be an etiologic factor of organic dust toxic syndrome among secondary school students employed in the grain dust environment polluted with the spores of Aspergillus candidus. The results obtained show that the agricultural work environment is considerably polluted with the spores of mould fungi of allergenic and immunotoxic properties, which cause a high degree of health risk to people employed in agriculture. PMID- 11059409 TI - [Evaluation of occupational exposure to mineral oil]. AB - The results of measurements of mineral oil concentrations in the air of selected workposts in five plants, taken in 1996-99, are presented. Absorption spectrometry in IR was used for determining mineral oils. In order to collect oil mist on a glass filter a personal sampling or stationary method was used. The results obtained show that the level of exposure to mineral oils in Polish industry is rather low. In few cases only oil mist was present in the air in amounts exceeding the Polish MAC value of 5 mg/m3. It was also noted that the situation has improved during the recent years. PMID- 11059410 TI - [Preliminary assessment of occupational exposure to Glutaraldehyde in selected endoscopic workplaces]. AB - The effect of occupational exposure to glutaric aldehyde in 17 endoscopic laboratories of the Wroclaw region was assessed. The level of glutaraldehyde concentration was measured, and the survey was carried out in a group of 34 laboratory employees. In 11 laboratories MAC and STEL values were exceeded. Undesirable effects following the contact with glutaric aldehyde were noted in 28 subjects (82%). The skin, mucous membrane and respiratory system irritations predominated among the symptoms observed. The data show that glutaric aldehyde may induce irritation or exert an allergic effects in medical personnel. PMID- 11059411 TI - [Analysis of costs involved in activities of regional occupational health centers]. AB - In this paper the authors continue considering the financial economy in the selected Regional Occupational Health Centres (ROHC). The paper contains an analysis of costs involved in the implementation of the ROHC activities, which constitute one of the components of the financial economy of health care institutions. Due to analyses carried out thus far it was possible to infer conclusions regarding the level and structure of the ROHC financing. The cost analysis supplements the considerations on the financial sources, and illustrates at the same time how the financial means have been used by these facilities. The ROHC expenditures were analysed in two dimensions: in the kind dimension and the subject-object dimension. Single costs were also the subject of the analysis. The comparison of the results obtained allowed for formulating conclusions on the financial economy of the selected ROHC. However, these conclusions cannot be generalised in terms of all facilities as the relationship between the structure and level of the costs, and the structure and level of financing was not common. The relationship between the highest share of salaries in the ROHC expenditures and a high level of financial support provided by regional Governors in 1998 was most frequently observed. A number of considerable limitations in the analysis performed were discussed in the final section of the paper. PMID- 11059412 TI - [Programmed cell death as a biological function of electromagnetic fields at a frequency of (50/60 Hz)--review]. AB - Apoptosis as physiological or programmed cellular death, and necrosis as pathological cellular death are two ways by which cells die. Recent data confirm that oxygen free radicals can be the mediators of apoptosis via signalling pathways in the cell. External magnetic fields are known to affect radical pair recombination and they may increase the concentration of oxygen free radicals in living cells. Therefore, it has been suggested that physical agents, such as electromagnetic fields of power-line frequency, could exert an effect on apoptosis via signalling pathways and oxygen free radicals. PMID- 11059413 TI - [Metallothioneins as stressor proteins modulating the immune response]. AB - Matallothioneins are a group of cellular proteins whose synthesis increases considerably in conditions of stressful reactions to harmful environmental factors, especially to heavy metals. They are considered to be proteins of intracellular activity whose specific effect on the metal-dependent enzymes is revealed within the cells, as well as proteins which regulate the transcription process. Metallothioneins released from cells to extracellular fluids, for example due to the damage of membrane, may disclose non-specific effect which usually does not occur. In such conditions, metallothioneins may also affect other cells of tissues and organs placed far away from targets of heavy metal. They also play an important role in the regulation of normal vital functions of the cells in the immune system. In numerous cases, because of their unique chemical structure and biological properties, metallothioneins perform the function of immunomodulators. In the majority of cases, the modulatory effect is favourable for the body as a whole. However, in some cases, this effect is negative, for example it hinders the removal of potentially neoplastic cells from the body. PMID- 11059414 TI - [Reduction of dustiness at non-stationary work stations]. AB - Many industrial processes are responsible for chemical and dust pollution. As the quantities of the substances emitted remain unknown in most cases, the data on the effect of technological parameters on the emission rates are not available. Various technical and organisational projects aimed at reducing dust hazards in rooms and at workposts are listed. Methods of assessing emission rates of pollutants from machines to the workplace air, and the efficiency of local exhaust ventilation are discussed. Results of the study in this area for a manual mechanised tool grinder and a tool grinder machine are presented. PMID- 11059415 TI - [International workshop of experts on "How to Protect Workers' Health?"--June 30 July 1, 2000. Lodz]. PMID- 11059416 TI - Public health in the new millennium II: Social exclusion. PMID- 11059417 TI - Truisms or truth? PMID- 11059418 TI - UNICEF says domestic violence against women and girls still a global epidemic. United Nations Children's Fund. PMID- 11059419 TI - Preventing firearm trauma: the role of local public health department. PMID- 11059420 TI - Mercury report released. PMID- 11059422 TI - Public health law at CDC. PMID- 11059421 TI - WHO assesses world's health systems. PMID- 11059423 TI - Soft drink "pouring rights": marketing empty calories to children. AB - Healthy People 2010 objectives call for meals and snacks served in schools to contribute to overall diets that meet federal dietary guidelines. Sales in schools of foods and drinks high in calories and low in nutrients undermine this health objective, as well as participation in the more nutritious, federally sponsored, school lunch programs. Competitive foods also undermine nutrition information taught in the classroom. Lucrative contracts between school districts and soft drink companies for exclusive rights to sell one brand are the latest development in the increasing commercialization of school food. These contracts, intended to elicit brand loyalty among young children who have a lifetime of purchases ahead of them, are especially questionable because they place schools in the position of "pushing" soft drink consumption. "Pouring rights" contracts deserve attention from public health professionals concerned about the nutritional quality of children's diets. PMID- 11059424 TI - Children and war. PMID- 11059425 TI - Bioterrorism preparedness and local public health agencies: building response capacity. PMID- 11059426 TI - "If it bleeds it leads"? Attributes of TV health news stories that drive viewer attention. AB - OBJECTIVE: Health advocates increasing y use the news media to educate the public. However, little is known about what motivates individuals to pay attention to health news. This study investigated which characteristics of TV health news stories attract viewer interest. METHODS: The authors surveyed airport patrons, the audience of a public health symposium, and municipal jurors, asking which attributes of TV heath news stories encouraged interest and which attributes discouraged interest. The authors ranked mean responses and compared them using Spearman rank correlations, RESULTS: The rankings assigned by the three samples were highly correlated. Respondents reported being most attracted to health stories about personally relevant topics. Interestingly, they also reported that sensational story elements such as "showing a bloody or injured person" and "being action packed" did not substantially influence their attention. CONCLUSIONS: This study suggests that viewers, regardless of their level of health knowledge, value the same attributes in TV health news stories. Emphasizing the personal relevance of health topics appears to be a viable strategy to capture viewer interest. Conversely, the tendency of broadcast news to sensationalize stories may be distracting in the case of health news. PMID- 11059427 TI - High incidence of extra-intestinal infections in a Salmonella Havana outbreak associated with alfalfa sprouts. AB - OBJECTIVE: To determine a vehicle and point source for an outbreak of Salmonella Havana. METHODS: The authors conducted a case-control study and traceback investigation of 14 residents of California and four from Arizona with onsets of illness from Apr 15, 1998, to June 15, 1998, and Salmonella Havana infections with identical PFGE patterns. RESULTS: Seventeen of 18 patients were women. Seventeen were adults 20-89 years of age. Nine (50%) had diarrheal illness, 6 (33%) had urinary tract infections, 2 (11%) had sepsis, and one had an infected surgical wound after appendectomy. Four patients were hospitalized, and one died. Eating alfalfa sprouts was associated with S. Havana infection (OR = 10.0; 95% confidence interval 1.2, 83.1; P = 0.01). CONCLUSIONS: This outbreak resulted in a high incidence of extra-intestinal infections, especially urinary tract infections, and high morbidity. Raw alfalfa sprouts, often considered a safe "heath food," can be a source of serious foodborne disease outbreaks. PMID- 11059428 TI - Self-reported postwar injuries among Gulf War veterans. AB - OBJECTIVE: From September 1995 to May 1996, the authors conducted a telephone survey of Iowa military personnel who had served in the regular military or activated National Guard or Reserve during the Gulf War period. To assess the association between military service in a combat zone and subsequent traumatic injury requiring medical consultation, the authors analyzed veterans' interview responses. METHODS: Using data from the larger survey, the authors compared rates of self-reported postwar injuries requiring medical consultation in a sample of Iowa Gulf War veterans to the rates in a sample of Iowa military personnel who served at the same time, but not in the Persian Gulf. RESULTS: Of 3695 veterans, 605 (16%) reported a traumatic injury in the previous three months requiring medical consultation. Self-reported injuries were associated with service in the Persian Gulf (odds ratio 1.26; 95% confidence interval 1.02, 1.55). CONCLUSION: This finding is consistent with the results of earlier studies of traumatic injury mortality rates among war veterans. PMID- 11059429 TI - External collaboration and performance: North Carolina local public health departments, 1996. AB - OBJECTIVES: This study examined the extent to which local public health departments in North Carolina collaborated with other groups and organizations, the health problems on which they worked together, and the effect of external collaboration on health departments' performance on core public health functions. METHODS: The author mailed a questionnaire asking about interactions with city and county government agencies, boards of health, schools, nonprofits, physicians/private clinics, community health centers/migrant clinics, community members, citizens' groups, state and federal agencies, and universities to all of the directors of local public health departments in North Carolina. Sixty-four directors returned the questionnaire, for a response rate of 74.4%. RESULTS: Local public health departments most frequently interacted with boards of health, state agencies, community members, schools, city and county government agencies, and nonprofit agencies. Large majorities reported productive relationships with boards of health, state agencies, city and county government agencies, schools, nonprofit agencies, and hospitals. Greater frequency of interaction with several types of partners was associated with better performance. CONCLUSIONS: While questions exist about whether performance on core functions improves the community's health status, the results suggest that it is important for local public health departments to continue to build relationships with other organizations in the community. PMID- 11059430 TI - National surveillance for infection with Cryptosporidium parvum, 1995-1998: what have we learned? AB - OBJECTIVE: Infection with Cryptosporidium parvum generally causes a self-limiting diarrheal illness. Symptoms can, however, last for weeks and can be severe, especially in immunocompromised individuals. In 1994, the Council of State and Territorial Epidemiologists (CSTE) recommended that cryptosporidiosis be a nationally notifiable disease. Forty-seven states have made infection with C. parvum notifiable to the Centers for Disease Control and Prevention (CDC), and laboratories in the three remaining states report cases to state health departments, which may report them to the CDC. To see what the data show about patterns of infection, the authors reviewed the first four years of reports to the CDC. METHODS: The authors analyzed reports of laboratory-confirmed cases of cryptosporidiosis for 1995-1998. RESULTS: During 1995-1998, 11,612 laboratory confirmed cases of cryptosporidiosis were reported to the CDC. All ages and both sexes were affected. An increase in case reporting was observed in late summer during each year of surveillance for people <20 years of age. CONCLUSION: The first national data on laboratory-confirmed cryptosporidiosis cases, although incomplete, provide useful information on the burden of disease in the nation as well as provide baseline data for monitoring of future trends. PMID- 11059431 TI - Elevated blood lead levels among adults in Massachusetts, 1991-1995. AB - OBJECTIVE: Lead poisoning, the oldest recognized occupational disease, remains a danger for children and adults. Data collected for 664 cases reported to the Massachusetts Occupational Lead Registry in 1991-1995 were summarized in a 1998 state report. Here, the authors present some of the key findings from that report for a wider audience. METHODS: The authors summarize key findings of the 1998 state report. FINDINGS: Construction workers, in particular licensed deleaders and house painters, accounted for almost 70% of occupational cases involving blood lead levels > or = 40 micrograms of lead per deciliter (mcg/dl) of blood. Among 100 workers with the highest blood lead levels (> or = 60 mcg/dl), 29% were house painters. Hispanic workers were over-represented in the Registry. A small proportion of cases were non-occupational, typically associated with recreational use of firing ranges or do-it-yourself home renovations. CONCLUSION: Lead poisoning is a preventable disease, yet these data indicate that additional prevention efforts are warranted. PMID- 11059432 TI - The health agency training program: continuing education courses in biostatistics and epidemiology. AB - The authors describe the development and evaluation of a continuing education program in biostatistics and epidemiology. Short courses were presented to public health and mental health professionals using teaching strategies that included lecture, discussion, practice-oriented examples, and interactive problem-solving. A total of 1723 health professionals attended one or more of the 120 courses presented from 1992 to 1996 in seven US states. Most course participants were female: the highest education level for 40% was a bachelor's degree, while 42% had advanced degrees. Approximately 66% of participants signed up for continuing education credits. The program represents a successful partnership between an academic institution and health agencies in a seven-state region. PMID- 11059433 TI - A university-public health practice linkage in Washington State. PMID- 11059434 TI - Excess risk of myocardial infarction in patients treated with antidepressant medications: association with use of tricyclic agents. AB - PURPOSE: Several studies have found that depression and the use of antidepressant medications are associated with an increased risk of cardiovascular disease. We assessed the association between the use of antidepressant drugs and myocardial infarction, and whether that association differs between the tricyclic and selective serotonin reuptake inhibitor (SSRI) classes of medication. PARTICIPANTS AND METHODS: We compared the experience of a cohort of 2,247 working, union health plan members who received at least one prescription for an antidepressant in an accrual period of 1991-1992 with that of 52,750 members who did not. Patients were followed for up to 4.5 years (minimum 6 months). Three antidepressant medication classes were defined: tricyclics, SSRIs, and others. The primary outcome was hospitalization or death due to myocardial infarction. RESULTS: Adjusted for age and sex, antidepressant users had a relative risk of myocardial infarction of 2.2 (95% confidence interval [CI] 1.3 to 3.7) compared with nonusers of antidepressants. There were 16 myocardial infarctions among 1,650 users of tricyclic antidepressants, 2 among 655 SSRI users, and none among 279 users of other antidepressants. Adjusting for age, gender, baseline heart disease, diabetes, hypertension, hyperlipidemia, anxiety, and cancer, the relative risk of myocardial infarction was 2.2 (95% CI 1.2 to 3.8) in users of tricyclic agents and 0.8 (95% CI 0.2 to 3.5) in users of SSRIs, as compared with subjects who did not use antidepressants. CONCLUSION: The association between use of tricyclic antidepressants, but not SSRIs, with an increased risk of myocardial infarction in our patients suggests that an earlier report that there is no difference in risk between the antidepressant classes, based on short-term studies, may not apply to long-term adverse cardiovascular outcomes. PMID- 11059435 TI - Nonalcoholic steatohepatitis and cryptogenic cirrhosis within kindreds. AB - PURPOSE: Familial forms of cryptogenic cirrhosis have been described. We have cared for families in which several members were afflicted with cryptogenic cirrhosis as well as the more recently recognized entity of nonalcoholic steatohepatitis. To examine the familial patterns of these disorders, we reviewed patients with nonalcoholic steatohepatitis, with and without cirrhosis, or cryptogenic cirrhosis to assess how frequently their relatives were afflicted with these disorders. SUBJECTS AND METHODS: Eighteen members of eight kindreds containing 2 or more afflicted members were studied. Diagnoses were based on histology in all but 3 patients (2 elderly women with liver atrophy and severe cirrhotic ascites diagnosed clinically with cryptogenic cirrhosis and 1 adult man with abnormal serum aminotransferase levels and hepatomegaly that was diagnosed as fatty liver by ultrasound). Other forms of liver disease were excluded by extensive serologic testing. RESULTS: There were 8 index patients (1 man, 7 women; 2 with cryptogenic cirrhosis, 4 with nonalcoholic steatohepatitis with cirrhosis, and 2 with nonalcoholic steatohepatitis without cirrhosis) and 10 relatives (4 men, 6 women; 2 with cryptogenic cirrhosis and 8 with nonalcoholic steatohepatitis). Nonalcoholic steatohepatitis and nonalcoholic steatohepatitis with cirrhosis coexisted within four kindreds, one of which also had an afflicted member with cryptogenic cirrhosis. Nonalcoholic steatohepatitis and cryptogenic cirrhosis coexisted within three additional kindreds. Patterns of afflicted patients included mother-daughter, sister-sister, sister-brother, father daughter, and male-female cousins. Fifteen (83%) of the 18 subjects were obese, and 11 (61%) had type 2 diabetes mellitus. CONCLUSIONS: The coexistence of nonalcoholic steatohepatitis with and without cirrhosis and cryptogenic cirrhosis within these kindreds suggests a common pathogenesis and possible genetic risk. These disorders were frequently but not invariably associated with female sex, obesity, and type 2 diabetes. PMID- 11059436 TI - Patient race and decisions to withhold or withdraw life-sustaining treatments for seriously ill hospitalized adults. SUPPORT Investigators. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - PURPOSE: Patient race is associated with decreased resource use for seriously ill hospitalized adults. We studied whether this difference in resource use can be attributed to more frequent or earlier decisions to withhold or withdraw life sustaining therapies. SUBJECTS AND METHODS: We studied adults with one of nine illnesses that are associated with an average 6-month mortality of 50% who were hospitalized at five geographically diverse teaching hospitals participating in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). We examined the presence and timing of decisions to withhold or withdraw ventilator support and dialysis, and decisions to withhold surgery. Analyses were adjusted for demographic characteristics, prognosis, severity of illness, function, and patients' preferences for life-extending care. RESULTS: The mean (+/- SD) age of the patients was 63 +/- 16 years; 16% were African-American, 44% were women, and 53% survived for 6 months or longer. Of the 9,076 patients, 5,349 (59%) had chart documentation that ventilator support had been considered in the event the patient's condition required such a treatment to sustain life, 2,975 charts (33%) had documentation regarding major surgery, and 1,293 (14%) had documentation of discussions about dialysis. There were no significant differences in the unadjusted rates of decisions to withhold or withdraw treatment among African-Americans compared with non-African-Americans: among African-Americans, 33% had a decision made to withhold or withdraw ventilator support compared with 35% among other patients, 14% had a decision made to withhold major surgery compared with 12% among other patients, and 25% had a decision made to withhold or withdraw dialysis compared with 30% among other patients (P >0.05 for all comparisons). After adjustment for demographic characteristics, prognosis, illness severity, function, and preferences for care, there were no differences in the timing or rate of decisions to withhold or withdraw treatments among African-Americans compared with non-African-American patients. CONCLUSION: Patient race does not appear to be associated with decisions to withhold or withdraw ventilator support or dialysis, or to withhold major surgery, in seriously ill hospitalized adults. PMID- 11059437 TI - Do automated calls with nurse follow-up improve self-care and glycemic control among vulnerable patients with diabetes? AB - PURPOSE: We sought to evaluate the effect of automated telephone assessment and self-care education calls with nurse follow-up on the management of diabetes. SUBJECTS AND METHODS: We enrolled 280 English- or Spanish-speaking adults with diabetes who were using hypoglycemic medications and who were treated in a county health care system. Patients were randomly assigned to usual care or to receive an intervention that consisted of usual care plus bi-weekly automated assessment and self-care education calls with telephone follow-up by a nurse educator. Outcomes measured at 12 months included survey-reported self-care, perceived glycemic control, and symptoms, as well as glycosylated hemoglobin (Hb A1c) and serum glucose levels. RESULTS: We collected follow-up data for 89% of enrollees (248 patients). Compared with usual care patients, intervention patients reported more frequent glucose monitoring, foot inspection, and weight monitoring, and fewer problems with medication adherence (all P -0.03). Follow-up Hb A,, levels were 0.3% lower in the intervention group (P = 0.1), and about twice as many intervention patients had Hb A1c levels within the normal range (P = 0.04). Serum glucose levels were 41 mg/dL lower among intervention patients than usual care patients (P = 0.002). Intervention patients also reported better glycemic control (P = 0.005) and fewer diabetic symptoms (P <0.0001 ), including fewer symptoms of hyperglycemia and hypoglycemia. CONCLUSIONS: Automated calls with telephone nurse follow-up may be an effective strategy for improving self-care behavior and glycemic control, and for decreasing symptoms among vulnerable patients with diabetes. PMID- 11059438 TI - Outcomes of care and resource utilization among patients with knee or shoulder disorders treated by general internists, rheumatologists, or orthopedic surgeons. AB - PURPOSE: Previous studies have suggested that specialists may achieve better clinical outcomes for patients, albeit often at greater cost. We sought to compare outcomes of care and resource utilization among patients with shoulder or knee problems who were treated by general internists, rheumatologists, and orthopedic surgeons. SUBJECTS AND METHODS: Outpatients with knee or shoulder complaints who were seen by general internists, rheumatologists, or orthopedic surgeons at an academic medical center were administered questionnaires at enrollment in the study and again 3 months later. The questionnaires included validated measures of satisfaction, functional status, and pain severity, as well as resource utilization. We compared baseline clinical characteristics, satisfaction with care, resource utilization, and changes in function and symptoms during 3 months of follow-up among patients who were cared for by the three different types of providers. RESULTS: A total of 534 patients responded to the baseline survey and 436 (82%) to the 3-month follow-up survey. About 60% (n = 323) had knee pain. Orthopedists cared for 40% (n = 211) of the patients, with the remainder treated in approximately equal numbers by general internists or rheumatologists. At baseline, patients of internists had less severe pain (differences of 0.3 to 0.6 points on a 1 to 5 scale, P <0.05) and functional limitations (differences of 0.4 to 0.6 points on a 1 to 5 scale, P <0.0006) than patients of rheumatologists and orthopedic surgeons. Adjusting for baseline differences, there were no significant differences among provider groups in pain relief or functional improvement during follow-up. However, in adjusted analyses, patients with shoulder pain who were cared for by orthopedic surgeons were least satisfied with the office environment [adjusted mean (+/- SD) satisfaction score of 1.6 +/- 0.8 on a 1 to 4 scale for orthopedic surgeons vs 1.3 +/- 0.8 for rheumatologists and 1.4 +/- 0.8 for internists, P = 0.004]. Among patients with knee pain, those treated by rheumatologists and orthopedic surgeons were more satisfied with the doctor-patient interaction (adjusted mean satisfaction scores of 1.1 +/- 0.9 for rheumatologists and 1.2 +/- 0.7 for orthopedic surgeons on a 1 to 4 scale vs 1.4 +/- 0.8 for general internists, P = 0.003). Orthopedic surgeons obtained significantly more radiographs of the knee or shoulder and more magnetic resonance imaging scans of the knee. Rheumatologists performed significantly more aspirations or injection procedures. Among all patients, those treated by rheumatologists were most satisfied with the physician interaction, and those treated by orthopedic surgeons were most satisfied with treatment results. CONCLUSION: The relative benefits of specialist compared with generalist care for patients with knee or shoulder pain depend on the importance attached to resource utilization, patient satisfaction, and health outcomes. PMID- 11059439 TI - Incident stroke after discharge from the hospital with a diagnosis of atrial fibrillation. AB - PURPOSE: Atrial fibrillation is an important risk factor for stroke. We analyzed stroke risk over time in patients discharged from the hospital with a diagnosis of incident atrial fibrillation as compared with the risk of stroke in the Danish population. SUBJECTS AND METHODS: In a random sample of half of the Danish population, we identified 13,625 men and 13,577 women, aged 50 to 89 years, with a hospital diagnosis of atrial fibrillation and no prior diagnosis of stroke during 1980 to 1993. Data on other medical conditions were also available from 1977 to 1993, but medication data were not available. Patients were followed from the diagnosis of atrial fibrillation until the first diagnosis of stroke (nonfatal or fatal cerebral ischemic infarct and cerebral hemorrhage), death, or the end of 1993. The risk of stroke in these patients was compared with the risk in the Danish population using Poisson regression modeling to estimate relative risks (RR) and 95% confidence intervals (CI). RESULTS: For men with atrial fibrillation, the stroke rates increased by age, from 13 per 1,000 person-years in those ages 50 to 59 years, to 22 per 1,000 person-years in those ages 60 to 69 years, to 42 per 1,000 person-years in those ages 70 to 79 years, to 51 per 1,000 person-years in those ages 80 to 89 years. Age-specific stroke rates were similar in women with atrial fibrillation. Patients with a hospital diagnosis of atrial fibrillation had an increased risk of stroke (RR = 2.4; 95% CI, 2.3 to 2.5 in men and RR = 3.0; 95% CI, 2.9 to 3.2 in women) compared with the Danish population. Stroke risk was greatest during the first year after discharge and decreased thereafter. Hypertension, diabetes, and peripheral atherosclerosis were also associated with an increased risk of stroke among patients with atrial fibrillation. Ischemic heart disease and heart failure were risk factors in men only. There was no reduction in the risk of stroke from 1980 to 1993. CONCLUSIONS: Men and women with atrial fibrillation are at a substantially increased risk of stroke, particularly in the first year after the diagnosis. PMID- 11059440 TI - Redefining medical treatment in the management of unstable angina. AB - In 1994, the Agency for Health Care Policy and Research sponsored the development of guidelines for diagnosing and managing patients with unstable angina. Since their publication, several important developments have occurred. The prognostic value of biochemical assays for cardiac-specific troponins T and I have been shown in many studies. The possible role for C-reactive protein in determining prognosis deserves further investigation. Substantial clinical benefits have been obtained with intravenous inhibitors of the platelet glycoprotein (GP) IIb-IIIa receptor (abciximab, eptifibatide, tirofiban) and with one of the low-molecular weight heparins (enoxaparin). The therapeutic potential of other low-molecular weight heparins, direct thrombin inhibitors, and oral GP IIb-IIIa inhibitors remains to be clarified. On the basis of this evidence, consideration should be given to measuring serum levels of a cardiac troponin (either T or I) and using intravenous GP IIb-IIIa inhibitors and low-molecular-weight heparin in the standard management of patients with unstable angina. PMID- 11059441 TI - Efficacy of newer medications for treating depression in primary care patients. AB - PURPOSE: Several medications have recently been introduced for the treatment of depression. We reviewed the literature to summarize their efficacy in the treatment of depression in adult patients in primary care settings. METHODS: We searched the literature published from 1980 to January 1998 using the Cochrane Collaboration Depression Anxiety and Neurosis Group's specialized registry of 8,451 clinical trials, references from trials and 46 pertinent meta-analyses, and consultation with experts. We included randomized controlled trials of at least 6 weeks' duration that measured clinical outcomes and compared one of 32 newer medications with another newer antidepressant, an older antidepressant, a placebo, or a psychosocial intervention for the treatment of depressed patients in primary care settings. The primary outcome was response rate, defined as the proportion of patients experiencing a 50% or greater improvement in depressive symptoms. RESULTS: There were 28 randomized controlled trials involving 5,940 adult primary care patients with major depression, depression requiring treatment, dysthymia, or mixed anxiety depression. Newer agents, including selective serotonin re-uptake inhibitors, serotonin norepinephrine inhibitors, reversible inhibitors of monoamine oxidase, and dopamine antagonists, were usually compared with tricyclic agents. Average response rates were 63% for newer agents, 35% for placebo, and 60% for tricyclic agents. Newer agents were significantly more effective than placebo [risk ratio = 1.6; 95% confidence interval (CI), 1.2 to 2.1), but similar to tricyclic agents (risk ratio = 1.0; 95% CI, 0.9 to 1.1). Response rates were similar in the different types of depressive disorders, except that two small trials in frail older patients showed no significant effects of newer agents compared with placebo. Dropout rates as a result of adverse effects were 8% with newer agents and 13% with tricyclic agents (P <0.05). CONCLUSIONS: In primary care settings, newer antidepressants are more effective than placebo and have similar efficacy compared with tricyclic agents in the acute treatment of depression. Dropout rates as a result of adverse effects are lower with newer compared with tricyclic agents. Future studies should compare the effectiveness of different therapies among primary care patients with less severe depression and greater medical and psychiatric comorbidity. PMID- 11059442 TI - Treatment of functional gastrointestinal disorders with antidepressant medications: a meta-analysis. AB - BACKGROUND: Functional gastrointestinal disorders are common, accounting for up to 50% of gastroenterology referrals, and several randomized controlled trials have evaluated antidepressant therapy for their treatment. METHODS: We performed a meta-analysis of published, English-language, randomized clinical trials on the use of antidepressants for the treatment of patients with functional gastrointestinal disorders. RESULTS: Twelve randomized placebo-controlled trials of antidepressant treatment of functional gastrointestinal disorders were identified. One was excluded for using a combination of a tricyclic and neuroleptic agent. The medications included tricyclic antidepressants (amitriptyline [n = 3], clomipramine [n = 1], desipramine [n = 2], doxepin [n = 1], and trimipramine [n = 2]), and the antiserotonin agent, mianserin (n = 2). In addition, one trial compared two different antidepressants (mianserin and clomipramine) with placebo. Data were abstracted for the dichotomous outcome of symptom improvement in seven studies, and for the continuous variable of pain score in eight studies. The summary odds ratio for improvement with antidepressant therapy was 4.2 (95% confidence interval [CI]: 2.3 to 7.9), and the average standardized mean improvement in pain was equal to 0.9 SD units (95% CI: 0.6 to 1.2 SD units). On average 3.2 patients needed to be treated (95% CI: 2.1 to 6.5 patients) to improve 1 patient's symptom. CONCLUSION: Treatment of functional gastrointestinal disorders with antidepressants appears to be effective. Whether this improvement is independent of an effect of treatment on depression needs further evaluation. PMID- 11059443 TI - Apoptosis in rheumatic diseases. PMID- 11059444 TI - What's in a name? Public knowledge, attitudes, and experiences with antibiotic use for acute bronchitis. PMID- 11059445 TI - Antidepressant use and the risk of myocardial infarction. PMID- 11059446 TI - Examining specialty care. PMID- 11059447 TI - "Sputum aeruginosa": time to rewrite the textbooks on virus-induced airway inflammation. PMID- 11059448 TI - Alliance to unify academic internal medicine. PMID- 11059449 TI - Ipsilesional versus contralesional neglect depends on attentional demands. AB - Right hemisphere injuries often produce contralesional hemispatial neglect (CN). In contrast to CN, some patients with right hemisphere damage can also show so called ipsilesional neglect (IN). Previous reports found that patients tend to show IN on line bisection tasks but CN on other tasks such as target cancellation. To learn why these two tasks induce different spatial biases in patients with right hemisphere injury, conventional (i.e. solid) line bisection was compared with two novel bisection tasks consisting of horizontally aligned strings of characters. The subjects' task was to mark a target character that was at or closest to the true midpoint of the simulated line. Four of the 5 patients showed a dissociation whereby IN occurred for solid lines while CN was observed on character lines. The two patients assessed with an antisaccade paradigm showed a "visual grasp" for leftward stimuli. The present results suggest that neglect on line bisection may reflect two opposing forces, an approach behavior or "visual grasp" toward left hemispace and an attentional bias toward right hemispace. PMID- 11059450 TI - Psychophysical properties of line bisection and body midline perception in unilateral neglect. AB - Past research associated unilateral neglect with a systematic ipsilesional shift of the perceived position of the body midline; however, this was not confirmed by recent experiments. We used the constant stimuli method to control for potential artifacts intrinsic to the techniques used in previous studies. Body midline perception was measured in the visual and proprioceptive modalities in ten patients with left unilateral neglect, ten control patients and ten normal subjects and compared with a visual line bisection task, also using the constant stimuli method. Neglect patients showed a significant rightward bias in the line bisection task, but no consistent directional bias either in the proprioceptive or in the visual body midline task. These results clearly counter the association between neglect and an ipsilesional shift of the body midline. However, in the body midline tasks neglect patients made more errors in judgement on both sides of their subjective midline, both with respect to the control groups and with respect to the line bisection task. This may imply that a specific impairment of body-centered representations is indeed present and manifests as a non directional increase in response variability, rather than as a systematic shift. It is suggested that body- and object-related tasks (such as line bisection) may be processed by independent cognitive computations. This interpretation is discussed with reference to a recent neuroimaging study investigating the same kinds of tasks. PMID- 11059451 TI - Predicting combinations of left and right asymmetries. AB - This paper explains how combinations of asymmetries for pairs of laterality variables may be predicted. It shows that many pairs combine as expected by chance, plus the influence of the RS+ gene hypothesised by Annett (1978, 1985). These include: eye dominance with writing hand, with throwing hand, and with foot for kicking; nonright handedness with planum temporale asymmetry, with asymmetry of the parietal operculum, and the association between these two anatomical asymmetries. Handedness for writing and throwing, and hand and foot preferences are more strongly associated, suggesting the presence of an additional influence. The reliability of the present analyses was supported by replication, especially in findings for hand preferences for writing and throwing described by Gilbert and Wysocki (1992). The relative frequencies of discordant patterns of preference, LR (writing, throwing) versus RL, are a function of the frequencies of the two variables in the population. These differ between age groups and also with the classification of 'either' hand preferences. Patterns of preference for writing, throwing and eye-dominance (RRR, RRL etc) are related in an orderly manner to differences between the hands for peg moving time. The success of the present application of the RS theory strengthens the argument that individual differences for hand preference depend on a continuous distribution of asymmetry, not on the 'types' commonly assumed in the literature. PMID- 11059452 TI - Vision and laterality: does occlusion disclose a feedback processing advantage for the right hand system? AB - The main purpose of this study was to examine whether manual asymmetries could be related to the superiority of the left hemisphere/right hand system in processing visual feedback. Subjects were tested when performing single (Experiment 1) and reciprocal (Experiment 2) aiming movements under different vision conditions (full vision, 20 ms on/180 ms off, 10/90, 40/160, 20/80, 60/120, 20/40). Although in both experiments right hand advantages were found, manual asymmetries did not interact with intermittent vision conditions. Similar patterns of results were found across vision conditions for both hands. These data do not support the visual feedback processing hypothesis of manual asymmetry. Motor performance is affected to the same extent for both hand systems when vision is degraded. PMID- 11059453 TI - Profound retrograde amnesia following mild head injury: organic or functional? AB - This paper describes a 56 year old female patient (JJ) who suffered a minor head injury at work and presented with profound retrograde amnesia for both public events and autobiographical material spanning her entire life. In addition, she complained of word-finding difficulties and anterograde memory impairment and neuropsychological assessment found evidence of mild executive dysfunction. Neurological investigations (CT and EEG) were essentially normal although changes indicative of small vessel disease were noted on MRI brain scan. Various forms and aetiologies of remote memory loss were considered including, simulated, psychogenic and organic amnesia, but differential diagnosis proved difficult. It is proposed that criteria used in clinical practice to differentiate functional and organic complaints are limited and this may be because (1) both factors can be involved in the aetiology of amnesia, and (2) a similar underlying brain mechanism, such as a retrieval deficit could underlie many instances of organic and psychogenic amnesia. Future research, complemented by functional brain imaging, is needed to explore the nature of retrieval deficits. PMID- 11059454 TI - What the locus of brain lesion tells us about the nature of the cognitive defect underlying category-specific disorders: a review. AB - Different models have been proposed to account for the nature of the cognitive defects underlying category-specific disorders for living and non-living things. One model assumes that the living/non-living distinction is the by-product of a more basic dichotomy, contingent upon the different weighting that visuo perceptual and functional attributes have in the identification of members of these categories. A second model submits that evolutionary pressure resulted in the elaboration of dedicated neural mechanisms for the domains of living (animals and plants) and non-living (artefacts) things. A third model proposes that the different level of interconnections existing between perceptual and functional features in living and non living things may be more important than the weighting of these features. Each of these models makes implicit assumptions about the extent and the localization of brain lesions provoking category-specific disorders. However, it must also be considered that these disorders are heterogeneous in nature, resulting from defects located at the semantic, lexical or visual level. In the present review of the literature, we kept this distinction in mind in trying to analyze the neuroanatomical correlates of living and non-living disorders. Our findings showed that there is a correlation between the locus of lesion and the patterns of categorical impairment: (a) a bilateral injury to the antero-mesial and inferior parts of the temporal lobes was found in patients with a category-specific semantic impairment for living things; (b) a lesion of the infero-mesial parts of the temporo-occipital areas of the left hemisphere was found in a group of patients showing a specific lexical impairment for members of the 'plants' category; (c) an extensive lesion of the areas lying on the dorso-lateral convexity of the left hemisphere was found in patients with a category-specific semantic impairment for man-made artefacts. Taken together, these results seem to show that the category-specific disorder is crucially related to the kind of semantic information processed by the damaged areas. PMID- 11059455 TI - Confabulation in a patient with fronto-temporal dementia and a patient with Alzheimer's disease. AB - This paper describes two patients, O.I. and B.Y., with a confabulatory syndrome. O.I. was diagnosed with probable fronto-temporal dementia, whereas B.Y. met the criteria for probable Alzheimer's disease. O.I., but not B.Y., was impaired on tests of frontal/executive functions, and performed better than B.Y. on clinical tests of memory. Both patients confabulated in episodic/autobiographical memory tasks and in personal future planning tasks. B.Y. confabulated also in a semantic memory task. It is argued that the pattern of confabulation and the cognitive profile shown by the two patients is explained better by the hypothesis proposed by Dalla Barba and co-workers (Dalla Barba et al., 1997b) than by current theories of confabulation. PMID- 11059456 TI - Functional imaging of visual semantic processing in the human brain. AB - Previous neuroimaging studies have identified a large network of cortical areas involved in semantic processing in the human brain, which includes left occipito temporal and inferofrontal areas. Most studies, however, investigated exclusively the associative/functional semantic knowledge by using mainly words and/or language related tasks, and this factor may have contributed to the large left hemisphere superiority found in semantic processing and to the controversial involvement of left prefrontal structures. The present study investigates the neural basis of visual objects knowledge, accessed exclusively through pictorial information. Regional cerebral blood flow (rCBF) was assessed using positron emission tomography (PET) during 3 conditions in right-handed normal volunteers: resting with eyes closed, retrieval of semantic information related to visual properties of objects (real size), and visual categorization based on physical properties of the image. Confirming previous experiments and neuropsychological findings, most activations were found in left occipito-temporal areas during retrieval of visual semantic knowledge. The absence of any activation in the left prefrontal inferior cortex for visual semantic processing confirms recent observations which suggest that this region would not be involved in retrieval of visual semantic knowledge from living entities. Rather, such knowledge about visual properties of objects, situated closely to cortical regions mediating perception of the visual attributes, can be retrieved directly from these regions when visual images are used as entry level stimuli. PMID- 11059457 TI - Verbal-response and manual-response versions of the Milner Landmark task: normative data. AB - The paper reports normative data relative to a shortened form of two versions of the Milner Landmark task, involving verbal and manual response, respectively, which have been found to provide crucial information to discriminate perceptual from response bias in unilateral neglect. Normative data based on a large group of subjects were believed to be necessary because the Landmark task is held to be worth further investigation: (a) in comparison with other tasks devised for similar purposes, (b) in elucidating clinico-anatomical correlations, and (c) in planning selective remediation programmes. The results of the investigation provide criteria on the basis of which a patient's bias can be classified as being normal, borderline, or pathological. PMID- 11059458 TI - Performance of a 2D image-based anthropometric measurement and clothing sizing system. AB - Two-dimensional, image-based anthropometric measurement systems offer an interesting alternative to traditional and three-dimensional methods in applications such as clothing sizing. These automated systems are attractive because of their low cost and the speed with which they can measure size and determine the best-fitting garment. Although these systems have appeal in this type of application, not much is known about the accuracy and precision of the measurements they take. In this paper, the performance of one such system was assessed. The accuracy of the system was analyzed using a database of 349 subjects (male and female) who were also measured with traditional anthropometric tools and techniques, and the precision was estimated through repeated measurements of both a plastic mannequin and a human subject. The results of the system were compared with those of trained anthropometrists, and put in perspective relative to clothing sizing requirements and short-term body changes. It was concluded that image-based systems are capable of providing anthropometric measurements that are quite comparable to traditional measurement methods (performed by skilled measurers), both in terms of accuracy and repeatability. PMID- 11059459 TI - Conditions for maintaining ageing operators at work--a case study conducted at an automobile manufacturing plant. AB - This is a case study on work/ageing relations in an automobile manufacturing company, where demographic trends and the work organisation determine the conditions under which ageing operators can work in repetitive tasks under tight time constraints. The methodology used is based on a comparison of different age operators at their regular workstation and in a new job, during the training process. The older workers appear to develop health-preserving strategies of work while achieving production goals. The possibility of setting up such strategies depends on the job characteristics. More generally, this case study allows us to better understand why some older workers are excluded from certain workstations, and thus why "polyvalency" or job rotation decreases with age. PMID- 11059460 TI - Sociotechnical principles for system design. AB - This paper offers a set of sociotechnical principles to guide system design, and some consideration of the role of principles of this kind. The principles extend earlier formulations by Cherns (1976, Human Relations, 29, 783-792; 1987, Human Relations, 40, 153-162). They are intended to apply to the design of new systems, including those incorporating new information technologies and a range of modern management practices and ways of working. They attempt to provide a more integrated perspective than is apparent in existing formulations. The principles are of three broad types: meta, content and process, though they are highly interrelated. They are for use by system managers, users and designers, and by technologists and social scientists. They offer ideas for debate and provide devices through which detailed design discussions can be elaborated. The principles are most likely to be effective if they are relatively freestanding, but supported by relevant methods and tools. The principles are necessary but not sufficient to make a substantial contribution to design practice. PMID- 11059461 TI - Voluntary pacing and energy cost of off-road cycling and running. AB - PURPOSE: The purposes of this study were (1) to compare self-chosen speed of off road cyclists and runners on a hilly course, (2) to compare the energy expenditure of off-road cyclists and runners on the same terrain, and (3) to describe changes in energy expenditure over the course of the exercise period. METHODS: Runners and cyclists performed three laps on a 2.75 km gravel course in a single exercise bout. The course was divided into 13 segments differing in grade and length. Position on the course and heart rate were recorded every few seconds. Speed was computed for each course segment on each lap; energy expenditure was estimated using recorded heart rates and exercise-specific maximal oxygen uptake measurements made prior to participation in the study. RESULTS: There were significant relationships between grade and speed for both runners (r = 0.64) and cyclists (r = 0.44). The differences between cyclists and runners were greatest on downhill segments. Energy expenditure rates were not significantly different for runners (71.6% VO2 peak) and cyclists (68.5% VO2 peak). CONCLUSIONS: Off-road cycling and running are comparable in energy demands. Variation in skill levels may account for the increased variability in speed among cyclists on downhill terrain. PMID- 11059462 TI - The influence of bed firmness on sleep quality. AB - We studied the relationship between sleep quality and bed surface firmness. Nine men were investigated, sleeping in their homes for at least 5 consecutive nights on a soft and a more firm mattress using a sensor pad placed under the mattress and a solid-state recording device. The subjective feeling of sleep quality did not always agree with the recorded sleep data. The difference was most marked when changing from the subject's own to one of the test mattresses. For the same subject the results were reproducible between nights provided there were no external disturbing factors. Four of the 9 subjects slept significantly better on the softer of the two mattresses and 2 on the hard mattress. The difference in sleep quality observed among the subjects tested makes it necessary to relate the results to the same person rather than considering a whole group as an entity. The adaptation period for a new sleep surface extended to many days. PMID- 11059463 TI - Assessment, re-design and evaluation of changes to the driver's cab in a suburban electric train. AB - A new type of electric train, called the Tangara, began replacing older trains on the Sydney city and suburban network in 1990. Shortly afterwards, some of the train drivers began reporting pain in the arms while driving the new train. The Ergonomics Unit of Worksafe Australia was then engaged to identify and assess ergonomic problems in the driver's cab. This process included direct observation of drivers at work, distribution of a self-administered questionnaire to all drivers, and analysis of anthropometric problems using a computer-aided design package. The analysis of 193 completed questionnaires and the problems shown by the design study were used in developing an improved design. A mock-up of the modified cab was made and tried out by 134 drivers whose comments led to further changes. The modified design was applied to new production, and existing cabs were modified during major maintenance. By 1996, 96% of the driver's cabs were to the new design. An evaluation of the new design was undertaken using written questionnaires, which were completed by 227 drivers. The results were strongly in favour of the new design, showing the effectiveness of the ergonomic modifications. PMID- 11059464 TI - A comparison of different postures for scaffold end-frame disassembly. AB - Overexertion and fall injuries comprise the largest category of nonfatal injuries among scaffold workers. This study was conducted to identify the most favourable scaffold end-frame disassembly techniques and evaluate the associated slip potential by measuring whole-body isometric strength capability and required coefficient of friction (RCOF) to reduce the incidence of injury. Forty-six male construction workers were used to study seven typical postures associated with scaffold end-frame disassembly. An analysis of variance (ANOVA) showed that the isometric forces (334.4-676.3 N) resulting from the seven postures were significantly different (p < 0.05). Three of the disassembly postures resulted in considerable biomechanical stress to workers. The symmetric front-lift method with hand locations at knuckle height would be the most favourable posture; at least 93% of the male construction worker population could handle the end frame with minimum overexertion risk. The static RCOF value resulting from this posture during the disassembly phase was less than 0.2, thus the likelihood of a slip should be low. PMID- 11059465 TI - Effects of semi-rigid arch-support orthotics: an investigation with potential ergonomic implications. AB - For many years, arch-support orthotics have been prescribed for individuals with discomfort and/or abnormal skeletal alignments in the structures of the lower extremity. Recently there has been an increased interest in promoting semi-rigid orthotics as an ergonomic aid for asymptomatic workers who must stand all day at their workplace. A laboratory study was performed to assess the biomechanical impact of prefabricated semi-rigid orthotics on asymptomatic individuals. Ten subjects wore semi-rigid arch-support orthotics (experimental condition) for two months and flexible polyurethane/Sorbothane shoe inserts (control condition) for two months. Throughout this 18-week testing period, the subjects returned to the lab to perform a battery of assessment tests at regularly scheduled intervals. These tests examined subject strength, standing posture, stability, fatigue effects, and body part discomfort. The results of this study showed no significant changes in the strength, posture, or stability as a function of insert type. The subjects reported a reduction in low-back discomfort along with an increase in foot discomfort during a fatiguing exertion task while wearing the semi-rigid orthotics as compared to the control condition. PMID- 11059466 TI - Visibility of road hazards in thermal, visible, and sensor-fused night-time imagery. AB - Sensor fusion combines the output of multiple imaging sensors within a single composite display. Ideally, a fused image will retain important spatial information provided by individual input images, and will convey useful spatial or chromatic emergent information derived from the contrast between input images. The present experiment assessed the potential benefits of sensor fusion as a method of enhancing drivers' night-time detection of road hazards. Observers were asked to detect a pedestrian within thermal and visible images of a night-time scene, and within chromatic and achromatic renderings created by sensor fusion of grayscale thermal and visible images. Results indicated that fusion can both improve spatial image content, and can effectively embellish spatial content with emergent chromatic information. The benefits of both sensor fusion and of color rendering, however, were inconsistent, varying substantially with quality of input images submitted for fusion. PMID- 11059467 TI - The influence of speed, grade and mass during simulated off road bicycling. AB - The purpose of this investigation was to examine the effects of bicycle mass, speed, and grade on oxygen consumption (VO2), heart rate (HR), and ratings of perceived exertion (RPE) during a simulated off-road riding paradigm. Nine adult subjects with mean +/- SD age, mass, and VO2 max of 26.1 +/- 5.6 years, 71.7 +/- 7.5 kg, 56.6 +/- 5.2 ml x kg(-1) x min(-1) respectively, were trained to ride a fully suspended Trek Y-22 mountain bike on a treadmill with a 3.8 cm bump affixed to the belt. Riders completed a maximum of nine separate trials encompassing three different bike masses (11.6, 12.6 and 13.6 kg), 3 speeds (2.7, 3.6 and 4.5 m x s(-1)), and 3 grades (0, 2.5, and 5%). Throughout a trial, bike mass and speed remained constant while riding grade was increased every 5 min. During simulated off-road riding on a fully suspended mountain bike, increases in speed and grade significantly increased VO2, heart rate, and RPE. Increases in bike mass had no significant effects on VO2, heart rate or RPE. In addition, speed and grade changes interacted to differentially affect VO2, heart rate, and RPE at all speeds and grades. PMID- 11059468 TI - The effects of interruptions in work activity: field and laboratory results. AB - The effects of interruptions in work activity were investigated, first in a field study where the operators' task was to card-index data about customers' phone lines. The interruptions due to customers' calls resulted in an increase of the processing time of the current task and in the use of several management strategies. A laboratory study was then designed in order to study the effects of temporal strain, complexity and similarity on time-sharing efficiency and to clarify the psychological mechanisms underlying the switching from one task to the other. The results showed especially a significant effect of temporal strains on performance and a strong increase in mean error rate at the very beginning of the processing of the second task. In conclusion, advice is given for both technical and organisational design. PMID- 11059469 TI - On the confusion between static load level and static task. PMID- 11059470 TI - Optimal gestational age and birth-weight cutoffs to predict neonatal morbidity. AB - BACKGROUND: Gestational age (GA) and birth weight (BW) criteria are used to identify newborns at risk for neonatal morbidity. Currently, preterm is GA less than 37 weeks; low birth weight is BW less than 2,500 grams; and small for gestational age (SGA) is BW less than the tenth percentile weight for the infant's GA. The optimal classification system balances the misclassification cost of false negatives against the cost of false positives. OBJECTIVE: To calculate the relative misclassification costs implied by the current 37-week and 2,500-gram cutoffs, and to test the validity of the current definition of SGA as a predictor of term morbidities. METHODS: GA, BW, and morbidity information were collected for 22,606 infants born between July 1981 and December 1992. Using this dataset, logistic regression coefficients were obtained modeling GA or BW as predictors of morbidities associated with prematurity. For a subset of 18,813 infants with GAs between 37 and 41 weeks, coefficients were obtained modeling both GA and BW as independent predictors of term morbidities. The logistic regression coefficients were used to calculate optimal birth weight, gestational age, and birth-weight-for-gestational-age cutoffs. RESULTS: The current definitions of low birth weight and preterm imply that it is 18 to 28 times more costly to misclassify a sick infant as low-risk than to misclassify a well infant as high-risk. CONCLUSIONS: Gestational age alone is better than birth weight alone at predicting preterm morbidities. No birth-weight cutoff can adequately predict term morbidities. A single weight-percentile cutoff for all gestational ages should not be used to identify newborns at high risk for neonatal morbidity. PMID- 11059471 TI - Early surgery versus conservative management of dissecting aneurysms of the descending thoracic aorta. AB - BACKGROUND: Optimal management for patients who present acutely with uncomplicated type III dissections of the descending thoracic aorta remains controversial. Patients with dissecting aneurysms represent a subgroup at high risk of rupture who may benefit from early elective surgery as an alternative to standard medical therapy. METHODS. The authors constructed a Markov decision model to compare the following clinical strategies: 1) early elective surgery immediately after diagnosis (EARLY SURGERY), 2) medical therapy with periodic computed tomography and with elective surgery when aortic diameter is projected to reach 6 cm (CT FOLLOW-UP), and 3) medical therapy with urgent surgery for dissection-related complications (WATCHFUL WAITING). Data sources included Medline (1966-1995) and a case series of patients with type III dissecting aneurysms who received medical therapy with radiographic follow-up. RESULTS: For a typical 60-year-old patient with an acute, uncomplicated 5-cm dissecting aneurysm of the descending thoracic aorta and an operative 30-day mortality rate of 14% for EARLY SURGERY, the model predicts that EARLY SURGERY improves survival compared with CT FOLLOW-UP (9.91 vs 9.44 QALYs). Conservative management may be preferred for patients who have maximum aneurysm diameters < or = 4 cm, are elderly (> or = 75 years), or have higher-than-expected risk of operative mortality. CONCLUSIONS: The choice between early surgery and medical therapy for uncomplicated dissecting aneurysm of the descending thoracic aorta should be tailored to the individual patient's operative risk, risk of dissection-related events, and age. Early surgery may be a reasonable alternative to medical therapy for carefully selected patients at centers with favorable perioperative mortality rates. PMID- 11059472 TI - Patient preference-based treatment thresholds and recommendations: a comparison of decision-analytic modeling with the probability-tradeoff technique. AB - BACKGROUND: Decision analysis (DA) and the probability-tradeoff technique (PTOT) are patient preference-based methods of determining optimal therapy for individuals. Using aspirin therapy for the primary prevention of stroke and myocardial infarction (MI) in elderly persons as an example, the objective of this study was to determine whether group-level treatment thresholds and individual-level treatment recommendations derived using PTOT are identical to those of DA incorporating the patients' own values. METHODS: Persons in a pilot study of the efficacy of aspirin in the prevention of stroke and MI were asked to participate. Participant values and utilities for pertinent health states (e.g., minor and major stroke, MI, major bleeding episode) were determined. Then, in three hypothetical clinical situations in which the chance of stroke or MI was varied, PTOT was used to directly determine treatment thresholds for aspirin therapy (i.e., the smallest reduction in MI or stroke risk for which participants would be willing to take aspirin). Using DA modeling, with the same probabilities of events as in the PTOT exercise and incorporating participants' own values, treatment thresholds for the three clinical situations were determined. The thresholds determined by the two approaches were compared. Finally, based on these treatment thresholds, using the best estimates of the efficacy of aspirin to prevent first-time stroke and MI, PTOT and DA treatment recommendations for individual participants were compared. RESULTS: The 42 participants reported that a major stroke was the least desirable health state, followed by MI, minor stroke, and major bleeding. The minimum risk reduction required to take aspirin was greater for MI prevention compared with stroke prevention. For the two clinical situations in which the hypothetical efficacy of aspirin to prevent stroke was varied, treatment thresholds for the PTOT versus DA approaches differed (p < 0.04), but this difference was not significant (p = 0.19) for the MI-based clinical situation. Using the best estimate of the efficacy of aspirin to prevent first-time stroke and MI, PTOT and DA treatment recommendations whether or not to take aspirin were discordant for 38% of participants (16 of 42) (p < 0.001). CONCLUSIONS: Patient preference-based group-level treatment thresholds and individual-level treatment recommendations can differ significantly depending on whether PTOT or DA is used, apparently because the two emphasize different aspects of the decision-making process. DA theory assumes that effective therapeutic decision making should maximize both quality and quantity of life; with PTOT, the emphasis for effective clinical decision making allows patients to be fully engaged in the process, thus hopefully leading to fully informed decisions that may result in satisfaction and compliance. PMID- 11059473 TI - Decision analysis with cumulative prospect theory. AB - BACKGROUND: Individuals sometimes express preferences that do not follow expected utility theory. Cumulative prospect theory adjusts for some phenomena by using decision weights rather than probabilities when analyzing a decision tree. METHODS: The authors examined how probability transformations from cumulative prospect theory might alter a decision analysis of a prophylactic therapy in AIDS, eliciting utilities from patients with HIV infection (n = 75) and calculating expected outcomes using an established Markov model. They next focused on transformations of three sets of probabilities: 1) the probabilities used in calculating standard-gamble utility scores; 2) the probabilities of being in discrete Markov states; 3) the probabilities of transitioning between Markov states. RESULTS: The same prophylaxis strategy yielded the highest quality adjusted survival under all transformations. For the average patient, prophylaxis appeared relatively less advantageous when standard-gamble utilities were transformed. Prophylaxis appeared relatively more advantageous when state probabilities were transformed and relatively less advantageous when transition probabilities were transformed. Transforming standard-gamble and transition probabilities simultaneously decreased the gain from prophylaxis by almost half. Sensitivity analysis indicated that even near-linear probability weighting transformations could substantially alter quality-adjusted survival estimates. CONCLUSION: The magnitude of benefit estimated in a decision-analytic model can change significantly after using cumulative prospect theory. Incorporating cumulative prospect theory into decision analysis can provide a form of sensitivity analysis and may help describe when people deviate from expected utility theory. PMID- 11059474 TI - A comparison of HUI2 and HUI3 utility scores in Alzheimer's disease. AB - PURPOSE: The Health Utilities Index (HUI) is a generic, multiattribute, preference-based health-status classification system. The HUI Mark 3 (HUI3) differs from the earlier HUI2 by modifying attributes and allowing more flexibility for capturing high levels of impairment. The authors compared HUI2 and HUI3 scores of patients with Alzheimer's disease (AD) and caregivers, and contrasted results of a cost-effectiveness analysis of new drugs for AD using the two systems. METHODS: In a cross-sectional study of 679 AD patient/caregiver pairs, stratified by patient's disease stage (questionable/mild/moderate/severe/profound/terminal) and setting (community/assisted living/nursing home), caregivers completed the combined HUI2/HUI3 questionnaire as proxy respondents for patients and for themselves. RESULTS: Mean (SD) global utility scores for patients were lower on the HUI3 (0.22[0.26]) than on the HUI2 (0.53 [0.21]). Patient HUI3 utility scores ranged from 0.47(0.24) for questionable AD to -0.23 (0.08) for terminal AD, compared with a range of 0.73 (0.15) to 0.14 (0.07) for the HUI2. Among the 203 patients in the severe, profound, and terminal stages, 96 (48%) had negative global HUI3 utility scores, while none had a negative HUI2 score. The utility scores for caregivers were similar on the HUI3 (0.87 [0.14]) and HUI2 (0.87 [0.11]). Cost effectiveness analysis of a new medication to treat AD showed somewhat more favorable results using the HUI3. CONCLUSIONS: The HUI2 and HUI3 discriminate well across AD stages. Compared with the HUI2, the HUI3 yields lower global utility scores for patients with AD, and more scores for states judged worse than dead. The HUI3 may yield substantially different results from the HUI2, particularly for persons who have serious cognitive impairments such as AD. PMID- 11059475 TI - Cardiopulmonary resuscitation preferences in Dutch community-dwelling and hospitalized elderly people: an interaction between gender and quality of life. AB - OBJECTIVE: To study the effects of information, gender, quality of life, and hospitalization on cardiopulmonary resuscitation (CPR) preferences and on the wish for information and participation in CPR discussions. METHODS: Seventy-five community-dwelling inhabitants of the city of Leiden and 45 consecutive patients in two hospitals in Leiden, The Netherlands, aged 75 years or older, were interviewed about their CPR preferences in their current states of health and in three hypothetical scenarios. Health-related quality of life (QOL) was assessed in separate items. The subjects were asked about their wishes for information and participation in CPR discussions. RESULTS: The chances of surviving CPR were overestimated. After receiving accurate information, 65% of the subjects, more women than men, did not want CPR. Overall QOL did not differ between men and women. Concerning the separate QOL items, men's CPR preferences were more associated with pain, whereas women's were more associated with being impaired in physical functioning and daily and social activities. CPR preferences in the current state of health did not differ significantly between community-dwelling and hospitalized participants. Although only 6% of all participants had ever discussed CPR with their doctors, 70% indicated they wanted routine CPR discussions (either when in good health at home or upon hospital admission), and 61% preferred to make the final decision about CPR themselves. CONCLUSIONS: CPR preferences are affected by different QOL items in men and women. CPR preferences in the current state of health do not differ between hospitalized and community dwelling elderly people. As the majority of elderly people want CPR discussions, they should be involved in decision making concerning CPR. PMID- 11059476 TI - Meta-analysis of ROC curves. AB - The authors present a method to combine several independent studies of the same (continuous or semiquantitative) diagnostic test, where each study reports a complete ROC curve; a plot of the true-positive rate or sensitivity against the false-positive rate or one minus the specificity. The result of the analysis is a pooled ROC curve, with a confidence band, as opposed to earlier proposals that result in a pooled area under the ROC curve. The analysis is based on a two parameter model for the ROC curve that can be estimated for each individual curve. The parameters are then pooled with a bivariate random-effects meta analytic method, and a curve can be drawn from the pooled parameters. The authors propose to use a model that specifies a linear relation between the logistic transformations of sensitivity and one minus specificity. Specifically, they define V = In(sensitivity/(1 - sensitivity)) and U = In((1 - specificity)/specificity), and then D = V - U, S = V + U. The model is defined as D = alpha + betaS. The parameters alpha and beta are estimated using weighted linear regression with bootstrapping to get the standard errors, or using maximum likelihood. The authors show how the procedure works with continuous test data and with categorical test data. PMID- 11059477 TI - Electronic trial banks: a complementary method for reporting randomized trials. AB - BACKGROUND: Randomized clinical trial (RCT) results are often difficult to find, interpret, or apply to clinical care. The authors propose that RCTs be reported into electronic knowledge bases-trial banks-in addition to being reported in text. What information should these trial-bank reports contain? METHODS: Using the competency decomposition method, the authors specified the ideal trial-bank contents as the information necessary and sufficient for completing the task of systematic reviewing. RESULTS: They decomposed the systematic reviewing task into four top-level tasks and 62 subtasks. 162 types of trial information were necessary and sufficient for completing these subtasks. These items relate to a trial's design, execution, administration, and results. CONCLUSION: Trial-bank publishing of these 162 items would capture into computer-understandable form all the trial information needed for critically appraising and synthesizing trial results. Decision-support systems that access shared, up-to-date trial banks could help clinicians manage, synthesize, and apply RCT evidence more effectively. PMID- 11059478 TI - A comprehensive league table of cost-utility ratios and a sub-table of "panel worthy" studies. AB - OBJECTIVES: The authors compiled a comprehensive league table of cost/QALY ratios, and a standardized table of analyses satisfying selected Reference Case criteria from the USPHS Panel on Cost-Effectiveness in Health and Medicine. METHODS: They identified 228 cost-utility analyses (CUAs) through literature searches, and abstracted data on methods and cost-utility ratios. The subset of "Panel-worthy" analyses used: a societal or broad health-care perspective, community or patient preference weights, net costs, incremental comparisons, and discounting of costs and QALYs. RESULTS: The 228 CUAs included ratios for 647 interventions, ranging from cost-saving to $52,000,000/QALY (median = $12,000/QALY). The standardized table presents 112 ratios that met the "Panel worthy" criteria, with articles published in recent years more likely to meet all of the criteria. CONCLUSIONS: The comprehensive league table (available on the Web) provides a useful reference, but ratios may not be comparable because of methodologic variations. The standardized table focuses on studies meeting basic methodologic criteria, potentially allowing for better comparison with future Reference Case analyses. Future studies should investigate the quality of analyses' underlying assumptions in addition to whether certain key procedural protocols were met. PMID- 11059479 TI - Determining the area under the ROC curve for a binary diagnostic test. AB - The authors provide a simple calculation for the unbiased estimation of the area under the ROC curve for a binary diagnostic test or a continuously valued test result that is effectively used in a binary way. The formula described can be used to interpret the discriminative ability of a diagnostic test. PMID- 11059480 TI - Law and ethics. PMID- 11059481 TI - Judgmental psychology. PMID- 11059482 TI - Use of green fluorescent protein color variants expressed on stable broad-host range vectors to visualize rhizobia interacting with plants. AB - We developed two sets of broad-host-range vectors that drive expression of the green fluorescent protein (GFP) or color variants thereof (henceforth collectively called autofluorescent proteins [AFPs]) from the lac promoter. These two sets are based on different replicons that are maintained in a stable fashion in Escherichia coli and rhizobia. Using specific filter sets or a dedicated confocal laser scanning microscope setup in which emitted light is split into its color components through a prism, we were able to unambiguously identify bacteria expressing enhanced cyan fluorescent protein (ECFP) or enhanced yellow fluorescent protein (EYFP) in mixtures of the two. Clearly, these vectors will be valuable tools for competition, cohabitation, and rescue studies and will also allow the visualization of interactions between genetically marked bacteria in vivo. Here, we used these vectors to visualize the interaction between rhizobia and plants. Specifically, we found that progeny from different rhizobia can be found in the same nodule or even in the same infection thread. We also visualized movements of bacteroids within plant nodule cells. PMID- 11059483 TI - Simultaneous imaging of Pseudomonas fluorescens WCS365 populations expressing three different autofluorescent proteins in the rhizosphere: new perspectives for studying microbial communities. AB - To visualize simultaneously different populations of pseudomonads in the rhizosphere at the single cell level in a noninvasive way, a set of four rhizosphere-stable plasmids was constructed expressing three different derivatives of the green fluorescent protein (GFP), namely enhanced cyan (ECFP), enhanced green (EGFP), enhanced yellow (EYFP), and the recently published red fluorescent protein (RFP; DsRed). Upon tomato seedling inoculation with Pseudomonas fluorescens WCS365 populations, each expressing a different autofluorescent protein followed by plant growth for 5 days, the rhizosphere was inspected using confocal laser scanning microscopy. We were able to visualize simultaneously and clearly distinguish from each other up to three different bacterial populations. Microcolonies consisting of mixed populations were frequently observed at the base of the root system, whereas microcolonies further toward the root tip predominantly consisted of a single population, suggesting a dynamic behavior of microcolonies over time. Since the cloning vector pME6010 has a broad host range for gram-negative bacteria, the constructed plasmids can be used for many purposes. In particular, they will be of great value for the analysis of microbial communities, for example in processes such as biocontrol, biofertilization, biostimulation, competition for niches, colonization, and biofilm formation. PMID- 11059484 TI - The sss colonization gene of the tomato-Fusarium oxysporum f. sp. radicis lycopersici biocontrol strain Pseudomonas fluorescens WCS365 can improve root colonization of other wild-type pseudomonas spp.bacteria. AB - We show that the disease tomato foot and root rot caused by the pathogenic fungus Fusarium oxysporum f. sp. radicis-lycopersici can be controlled by inoculation of seeds with cells of the efficient root colonizer Pseudomonas fluorescens WCS365, indicating that strain WCS365 is a biocontrol strain. The mechanism for disease suppression most likely is induced systemic resistance. P. fluorescens strain WCS365 and P. chlororaphis strain PCL1391, which acts through the production of the antibiotic phenazine-1-carboxamide, were differentially labeled using genes encoding autofluorescent proteins. Inoculation of seeds with a 1:1 mixture of these strains showed that, at the upper part of the root, the two cell types were present as microcolonies of either one or both cell types. Microcolonies at the lower root part were predominantly of one cell type. Mixed inoculation tended to improve biocontrol in comparison with single inoculations. In contrast to what was observed previously for strain PCL1391, mutations in various colonization genes, including sss, did not consistently decrease the biocontrol ability of strain WCS365. Multiple copies of the sss colonization gene in WCS365 improved neither colonization nor biocontrol by this strain. However, introduction of the sss-containing DNA fragment into the poor colonizer P. fluorescens WCS307 and into the good colonizer P. fluorescens F113 increased the competitive tomato root tip colonization ability of the latter strains 16- to 40-fold and 8- to 16-fold, respectively. These results show that improvement of the colonization ability of wild-type Pseudomonas strains by genetic engineering is a realistic goal. PMID- 11059485 TI - Bean dwarf mosaic virus BV1 protein is a determinant of the hypersensitive response and avirulence in Phaseolus vulgaris. AB - The capacities of the begomoviruses Bean dwarf mosaic virus (BDMV) and Bean golden yellow mosaic virus (BGYMV) to differeBean dwarf mosaic viru certain common bean (Phaseolus vulgaris) cultivars were used to identify viral determinants of the hypersensitive response (HR) and avirulence (avr) in BDMV. A series of hybrid DNA-B components, containing BDMV and BGYMV sequences, was constructed and coinoculated with BDMV DNA-A (BDMV-A) or BDMVA-green florescent protein into seedlings of cv. Topcrop (susceptible to BDMV and BGYMV) and the BDMV-resistant cvs. Othello and Black Turtle Soup T-39 (BTS). The BDMV avr determinant, in bean hypocotyl tissue, was mapped to the BDMV BV1 open reading frame and, most likely, to the BV1 protein. The BV1 also was identified as the determinant of the HR in Othello. However, the HR was not required for resistance in Othello nor was it associated with BDMV resistance in BTS. BDMV BV1, a nuclear shuttle protein that mediates viral DNA export from the nucleus, represents a new class of viral avr determinant. These results are discussed in terms of the relationship between the HR and resistance. PMID- 11059486 TI - Bromovirus movement protein conditions for the host specificity of virus movement through the vascular system and affects pathogenicity in cowpea. AB - Previously, we reported that CCMV(B3a), a hybrid of bromovirus Cowpea chlorotic mottle virus (CCMV) with the 3a cell-to-cell movement protein (MP) gene replaced by that of cowpea-nonadapted bromovirus Brome mosaic virus (BMV), can form small infection foci in inoculated cowpea leaves, but that expansion of the foci stops between 1 and 2 days postinoculation. To determine whether the lack of systemic movement of CCMV(B3a) is due to restriction of local spread at specific leaf tissue interfaces, we conducted more detailed analyses of infection in inoculated leaves. Tissue-printing and leaf press-blotting analyses revealed that CCMV(B3a) was confined to the inoculated cowpea leaves and exhibited constrained movement into leaf veins. Immunocytochemical analyses to examine the infected cell types in inoculated leaves indicated that CCMV(B3a) was able to reach the bundle sheath cells through the mesophyll cells and successfully infected the phloem cells of 50% of the examined veins. Thus, these data demonstrate that the lack of long distance movement of CCMV(B3a) is not due to an inability to reach the vasculature, but results from failure of the virus to move through the vascular system of cowpea plants. Further, a previously identified 3a coding change (A776C), which is required for CCMV(B3a) systemic infection of cowpea plants, suppressed formation of reddish spots, mediated faster spread of infection, and enabled the virus to move into the veins of inoculated cowpea leaves. From these data, and the fact that CCMV(B3a) directs systemic infection in Nicotiana benthamiana, a permissive systemic host for both BMV and CCMV, we conclude that the bromovirus 3a MP engages in multiple activities that contribute substantially to host-specific long-distance movement through the phloem. PMID- 11059487 TI - Saprophytic intracellular rhizobia in alfalfa nodules. AB - In indeterminate alfalfa nodules, the establishment of the senescent zone IV, in which both symbionts undergo simultaneous degeneration, has been considered, until now, as the end point of the symbiotic interaction. However, we now describe an additional zone, zone V, proximal to the senescent zone IV and present in alfalfa nodules more than 6 weeks old. In zone V, a new round of bacterial release occurs from remaining infection threads, leading to the reinvasion of plant cells that have completely senesced. These intracellular rhizobia are rod shaped and do not display the ultrastructural differentiation features of bacteroids observed in the more distal zones of the nodule. Interestingly, we have found that oxygen is available in zone V at a concentration compatible with both bacterial development and nitrogen fixation gene expression in newly released rhizobia. However, this expression is not correlated with acetylene reduction. Moreover, the pattern of nifH expression in this zone, as well as new data relating to expression in zone II, strongly suggest that nifH transcription in the nodule is under the control of a negative regulator in addition to oxygen. Our results support the conclusion that zone V is an ecological niche where intracellular rhizobia take advantage of the interaction for their exclusive benefit and live as parallel saprophytic partners. The demonstration of such an advantage for rhizobia in nodules was the missing evidence that Rhizobium-legume interactions are indeed symbiotic and, in particular, suggests that benefits to the two partners are associated with different developmental stages within the nodule. PMID- 11059488 TI - Site-directed mutagenesis of the magB gene affects growth and development in Magnaporthe grisea. AB - G protein signaling is commonly involved in regulating growth and differentiation of eukaryotic cells. We previously identified MAGB, encoding a Galpha subunit, from Magnaporthe grisea, and disruption of MAGB led to defects in a number of cellular responses, including appressorium formation, conidiation, sexual development, mycelial growth, and surface sensing. In this study, site-directed mutagenesis was used to further dissect the pleiotropic effects controlled by MAGB. Conversion of glycine 42 to arginine was predicted to abolish GTPase activity, which in turn would constitutively activate G protein signaling in magB(G42R). This dominant mutation caused autolysis of aged colonies, misscheduled melanization, reduction in both sexual and asexual reproduction, and reduced virulence. Furthermore, magB(G42R) mutants were able to produce appressoria on both hydrophobic and hydrophilic surfaces, although development on the hydrophilic surface was delayed. A second dominant mutation, magB(G203R) (glycine 203 converted to arginine), was expected to block dissociation of the Gbetagamma from the Galpha subunit, thus producing a constitutively inactive G protein complex. This mutation did not cause drastic phenotypic changes in the wild-type genetic background, other than increased sensitivity to repression of conidiation by osmotic stress. However, magB(G203R) is able to complement phenotypic defects in magB mutants. Comparative analyses of the phenotypical effects of different magB mutations are consistent with the involvement of the Gbetagamma subunit in the signaling pathways regulating cellular development in M. grisea. PMID- 11059489 TI - A guaB mutant strain of Rhizobium tropici CIAT899 pleiotropically defective in thermal tolerance and symbiosis. AB - Rhizobium tropici strain CIAT899 displays a high intrinsic thermal tolerance, and had been used in this work to study the molecular basis of bacterial responses to high temperature. We generated a collection of R. tropici CIAT899 mutants affected in thermal tolerance using TnS-luxAB mutagenesis and described the characterization of a mutant strain, CIAT899-10T, that fails to grow under conditions of high temperature. Strain CIAT899-10T carries a single transposon insertion in a gene showing a high degree of similarity with the guaB gene of Escherichia coli and other organisms, encoding the enzyme inosine monophosphate dehydrogenase. The guaB strain CIAT899-10T does not require guanine for growth due to an alternative pathway via xanthine dehydrogenase and, phenotypically, in addition to the thermal sensitivity, the mutant is also defective in symbiosis with beans, forming nodules that lack rhizobial content. Guanine and its precursors restore wild-type tolerance to grow at high temperature. Our data show that, in R. tropici, the production of guanine via inosine monophosphate dehydrogenase is essential for growth at extreme temperatures and for effective nodulation. PMID- 11059490 TI - Crg, a gene required for Ur-3-mediated rust resistance in common bean, maps to a resistance gene analog cluster. AB - Race-specific resistance to the bean rust pathogen (Uromyces appendiculatus) is provided by a number of loci in common bean (Phaseolus vulgaris). The Ur-3 locus controls hypersensitive resistance (HR) to 44 of the 89 races curated in the United States. To better understand resistance mediated by this locus, we developed new genetic material for analysis. We developed a population of mutagenized seed of cv. Sierra (genotype = Ur-3 ur-4 ur-6) that was screened with a bean rust race that is normally incompatible (HR response) on Ur-3 genotypes. We discovered two mutants of common bean, crg and ur3-delta3, in which uredinia formed on leaves (a compatible interaction) following infection. The F1 generation from a cross of these two mutants expressed the HR response, and the F2 generation segregated in a ratio of 9:7 (HR/uredinia formation). Therefore, the two genes are unlinked. Further genetic analysis determined that the mutation in ur3-delta3 was in the Ur-3 locus, and the mutation in crg was in a newly discovered gene given the symbol Crg (Complements resistance gene). Each mutation was inherited in a recessive manner. Unlike ur3-delta3, crg expressed reduced compatibility to bean rust races 49 and 47 that are normally fully compatible on genotypes, such as Sierra, that are homozygous recessive at the Ur-4 and Ur-6 loci. This suggests a gene mutated in crg is normally a positive compatibility factor for the bean-bean rust interaction. Polymerase chain reaction analysis of crg with primers to common bean resistance gene analogs (RGA) that contain a nucleotide-binding site sequence similar to those found in a number of plant disease resistance genes revealed that crg is missing the SB1 RGA, but not the linked SB3 and SB5 RGAs. Genetic analyses revealed that Crg cosegregates with the SB1 RGA. These results demonstrate that Crg is located near a RGA cluster in the common bean genome. PMID- 11059491 TI - Improved gfp and inaZ broad-host-range promoter-probe vectors. AB - A new set of broad-host-range promoter-probe vectors has been constructed. One subset contains the pVS1 and p15a replicons and confers resistance to either gentamicin or kanamycin. The other set contains the broad-host-range replicon from pBBR1 and confers resistance to kanamycin, tetracycline, ampicillin, or spectinomycin/streptomycin. Both plasmid sets are highly stable and are maintained without selection for more than 30 generations in several bacterial taxa. Each plasmid contains a promoter-probe cassette that consists of a multicloning site, containing several unique restriction sites, and gfp or inaZ as a reporter gene. The cassette is bound by transcriptional terminators to permit the insertion of strong promoters and to insulate the cassette from external transcription enabling the detection of weak or moderate promoters. The vector suite was augmented with derivatives of the kanamycin-resistant gfp promoter-probe plasmids that encode Gfp variants with different half-life times. PMID- 11059492 TI - Regulation of hrp genes and type III protein secretion in Erwinia amylovora by HrpX/HrpY, a novel two-component system, and HrpS. AB - Two novel regulatory components, hrpX and hrpY, of the hrp system of Erwinia amylovora were identified. The hrpXY operon is expressed in rich media, but its transcription is increased threefold by low pH, nutrient, and temperature levels- conditions that mimic the plant apoplast. hrpXY is autoregulated and directs the expression of hrpL; hrpL, in turn, activates transcription of other loci in the hrp gene cluster (Z.-M. Wei and S. V. Beer, J. Bacteriol. 177:6201-6210, 1995). The deduced amino -acid sequences of hrpX and hrpY are similar to bacterial two component regulators including VsrA/VsrD of Pseudomonas (Ralstonia) solanacearum, DegS/DegU of Bacillus subtilis, and UhpB/UhpA and NarX/NarP, NarL of Escherichia coli. The N-terminal signal-input domain of HrpX contains PAS domain repeats. hrpS, located downstream of hrpXY, encodes a protein with homology to WtsA (HrpS) of Erwinia (Pantoea) stewartii, HrpR and HrpS of Pseudomonas syringae, and other delta54-dependent, enhancer-binding proteins. Transcription of hrpS also is induced under conditions that mimic the plant apoplast. However, hrpS is not autoregulated, and its expression is not affected by hrpXY. When hrpS or hrpL were provided on multicopy plasmids, both hrpX and hrpY mutants recovered the ability to elicit the hypersensitive reaction in tobacco. This confirms that hrpS and hrpL are not epistatic to hrpXY. A model of the regulatory cascades leading to the induction of the E. amylovora type III system is proposed. PMID- 11059493 TI - Transposon insertion in the ftsK gene impairs in planta growth and lesion-forming abilities in Pseudomonas syringae pv. syringae B728a. AB - A Tn5 insertion in the ftsK gene of Pseudomonas syringae pv. syringae B728a impaired brown spot lesion formation on Phaseolus vulgaris, the ability to grow within bean leaves, and swarming ability on semisolid agar. Plasmids containing the ftsK gene were sufficient to complement the original Tn5 mutant for lesion formation and swarming and partially restored in planta growth. PMID- 11059494 TI - The I gene of bean: a dosage-dependent allele conferring extreme resistance, hypersensitive resistance, or spreading vascular necrosis in response to the potyvirus Bean common mosaic virus. AB - The resistance to the potyvirus Bean common mosaic virus (BCMV) conferred by the I allele in cultivars of Phaseolus vulgaris has been characterized as dominant, and it has been associated with both immunity and a systemic vascular necrosis in infected bean plants under field, as well as controlled, conditions. In our attempts to understand more fully the nature of the interaction between bean with the I resistance allele and the pathogen BCMV, we carefully varied both I allele dosage and temperature and observed the resulting, varying resistance responses. We report here that the I allele in the bean cultivars we studied is not dominant, but rather incompletely dominant, and that the system can be manipulated to show in plants a continuum of response to BCMV that ranges from immunity or extreme resistance, to hypersensitive resistance, to systemic phloem necrosis (and subsequent plant death). We propose that the particular phenotypic outcome in bean results from a quantitative interaction between viral pathogen and plant host that can be altered to favor one or the other by manipulating I allele dosage, temperature, viral pathogen, or plant cultivar. PMID- 11059495 TI - The dnaJ (hsp40) locus in Rhizobium leguminosarum bv. phaseoli is required for the establishment of an effective symbiosis with Phaseolus vulgaris. AB - P121R25 is a Tn5-induced mutant of the effective Rhizobium leguminosarum bv. phaseoli strain P121R that is unable to use glutamate as the sole carbon and nitrogen source and is defective in symbiotic nitrogen fixation. Enzymatic analysis showed that three enzymes implicated in glutamate metabolism (glutamate dehydrogenase, 2-oxoglutarate dehydrogenase, and glutamate synthase) were affected by this mutation. Sequencing of the chromosomal locus bordering the Tn5 in P121R25 indicated the presence of the dnaK and dnaJ genes in an arrangement similar to that described in R. leguminosarum bv. viciae (GenBank accession number Y14649). The mutation was located in the dnaJ (hsp40) gene. PMID- 11059496 TI - Agroinfiltration of Cauliflower mosaic virus gene VI elicits hypersensitive response in Nicotiana species. AB - Cauliflower mosaic virus strain W260 induces hypersensitive response (HR) in Nicotiana edwardsonii and systemic cell death in N. clevelandii. In contrast, the D4 strain of Cauliflower mosaic virus evades the host defenses in Nicotiana species; it induces chlorotic primary lesions and a systemic mosaic in both hosts. Previous studies with chimeric viruses had indicated that gene VI of W260 was responsible for elicitation of HR or cell death. To prove conclusively that W260 gene VI is responsible, we inserted gene VI of W260 and D4 into the Agrobacterium tumefaciens binary vector pKYLX7. Agroinfiltration of these constructs into the leaves of N. edwardsonii and N. clevelandii revealed that gene VI of W260 elicited HR in N. edwardsonii 4 to 5 days after infiltration and cell death in N. clevelandii approximately 9 to 12 days after infiltration. In contrast, gene VI of D4 did not elicit HR or cell death in either Nicotiana species. A frameshift mutation introduced into gene VI of W260 abolished its ability to elicit HR or cell death in both Nicotiana species, demonstrating that the elicitor is the gene VI protein. PMID- 11059497 TI - The way forward is to know the way back: the benefits of being adventurous. PMID- 11059498 TI - The cat sat on the mat: evolution of a scientific debate. PMID- 11059499 TI - The WHO forum on traditional medicine in health systems, Harare, Zimbabwe, February 14-18, 2000. PMID- 11059500 TI - Detection of antimicrobials in bee products with activity against viridans streptococci. AB - OBJECTIVES: Bee products have been studied extensively for their healing properties and have become part of cosmetic preparations and folk medicine. The major objective of this study was to examine the presence of antimicrobials in various bee products. DESIGN: Propolis, honeycomb lids, pollen, honeycombs, and honey were screened for antimicrobial compounds. Viridans streptococci were used as indicator strains. Ethanol extracts were applied onto paper disks, dried, and put on the surface of nutrient agar plates with the overlay containing viridans streptococci. The plates were then incubated and evaluated the next day for the presence of inhibition zones. The size of the inhibition zone represented a quantitative measure of antimicrobial activity in a sample. Thin-layer chromatography was used for separation of compounds in the samples and biodetection-an overlay with indicator strain-identified the antimicrobial compounds by formation of inhibition zones. RESULTS: Ethanol extracts of propolis and honeycomb lids, as well as honey containing honeycomb lids contained a mixture of antimicrobial compounds in various amounts. Thin-layer chromatography experiments with two different solvent systems differing in polarity suggested that major antimicrobials present in the samples prepared from honeycomb lids, honeycombs, pollen, and propolis have similar properties. These active compounds were not extremely hydrophobic. CONCLUSIONS: Extracts from the tested samples of bee products exhibited antimicrobial properties at various levels depending on the sample and bacterial species used for testing. Using chromatographic techniques, we demonstrated similar properties of the active compounds in various active samples. In conclusion, our results suggest that honey containing honeycomb lids may be a beneficial food supplement because of the presence of antibacterial compounds. PMID- 11059501 TI - Clinical evidence of subcutaneous CO2 insufflations: a systematic review. AB - OBJECTIVE: To evaluate the evidence of clinical effectiveness of subcutaneous CO2 insufflations (SCIs). STUDY LOCATION: The entire databases of MEDLINE, EMBASE Excerpta Medica (Field: Rehabilitation and Physical Medicine) and the Cochrane Library were screened for data up to August 1999. In addition, other potentially relevant journals were handsearched and references were checked. STUDY SELECTION: Uncontrolled observational trials (UCOTs) with sample sizes of at least n = 100, and controlled clinical trials (CCTs) and randomized controlled trials (RCTs) were included. Outcomes had to be patient-centered. English-, Czechoslovakian-, and German-language papers were considered. DATA ANALYSIS: For all study types, a criteria-based analysis was performed. For controlled trials only, quality was quantitatively assessed with the Jadad and Maastricht Scales. RESULTS: Thirteen studies (5 RCTs, 2 CCTs, and 6 UCOTs) were included. Mean scores on the Jadad and Maastricht Scales were 2 (Maximum 5) and 37 (Maximum 100), respectively. SCIs were found to be effective in addition to standard physical therapy in "peripheral arterial occlusive disease" (2 RCTs) and "stable angina pectoris" (1 RCT) and superior to sham needling for migraine headaches (1 RCT). There were no differences among SCIs and CO2 gas baths (1 RCT) or combined interventions, including SCIs (2 CCTs). All 6 UCOTs showed marked longitudinal effects. CONCLUSIONS: The low number and quality of the available studies precludes firm conclusions on the clinical effectiveness of SCIs. PMID- 11059502 TI - The effects of massage therapy alone and in combination with other complementary therapies on immune system measures and quality of life in human immunodeficiency virus. AB - OBJECTIVES: Determine effects of massage therapy alone and in combination with exercise or stress management-biofeedback treatment on enumerative immune measures, and quality of life in moderately immunocompromised human immunodeficiency virus (HIV) subjects. DESIGN: Randomized prospective controlled trial with 42 subjects randomly assigned to one of three treatment groups or a control group receiving standard care and intervention over a 12-week period. SETTING: Academic medical center. SUBJECTS: Forty-two (42) subjects with HIV infection (40 males; 2 females; aged 27-50 years) met eligibility requirements of CD4+ lymphocyte cell count greater than 200 cells per microliter; no present or recent signs or symptoms of acquired immunodeficiency syndrome (AIDS), and were not hospitalized. INTERVENTIONS: A 45-minute overall body massage once per week; similar massage and supervised aerobic exercise 2 other days per week; similar massage and biofeedback stress management once per week; control receiving standard treatment. OUTCOME MEASURES: Changes in peripheral blood levels of CD4+ lymphocytes, CD8+ lymphocytes, CD4+/CD8+ lymphocyte ratio and natural killer cells; six dimension quality-of-life assessment. RESULTS: No significant changes (p > 0.05) were found in any enumerative immune measure. Significant (p < 0.05) differences for quality-of-life assessment were in health care utilization and health perceptions, favoring massage and stress management compared to massage only and controls. CONCLUSIONS: Massage administered once per week to HIV infected persons does not enhance immune measures. Massage combined with stress management favorably alters health perceptions and leads to less utilization of health care resources. This suggests that HIV-infected persons receiving massage and stress management would tend to not overutilize health care services, thus possibly reducing health care costs. PMID- 11059503 TI - Use of complementary medicine by adult patients participating in HIV/AIDS clinical trials. AB - OBJECTIVE: To identify and characterize patterns of use of complementary and alternative (CAM) therapies by human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) patients participating in clinical trials in a research setting. DESIGN: A descriptive survey using a nonrandom sample of 100 patients was conducted over 17 months, using a 99-item interview schedule adapted from a previous study. SETTING: National Institutes of Health (NIH) Warren G. Magnuson Clinical Center. SUBJECTS: Patients diagnosed with HIV/AIDS, participating in clinical research protocols at the National Institute of Allergy and Infectious Diseases and the National Cancer institute (NCI) at the NIH. RESULTS: Ninety-one percent (91%) had used at least one CAM therapy, as defined by a 1993 study by Eisenberg et al., sometime in their lives. Postdiagnosis, 84% used at least one CAM therapy with an average of just fewer than 5. The increase in frequency of use from 64% prediagnosis was significant (P2 = 0.019). Therapies that became significantly more popular postdiagnosis were, imagery (P2 = 0.00012), high-dose vitamins, (P2 = 0.000019), weight gain (p2 = 0.000244), massage (p2 = 0.00091), relaxation (p2 = 0.0033), herbals (p2 = 0.013), spiritual (p2 = 0.024), and acupuncture (p2 = 0.035). They were primarily used for HIV/AIDS related problems: dermatological, nausea, depression, insomnia, and weakness. There was a high level of agreement that benefits of CAM use were: feeling better, 51 (98.1%), increased coping, 52 (100%), feeling in control, 44 (88.5%), and enhanced treatment outcome, 49 (94.2%) with 32 (61%) stating CAM was as, or more effective than conventional treatment. Fifty-three percent (53%) were specifically asked by physicians whether they were using adjunct therapies. CONCLUSIONS: Subjects used a variety of CAM therapies to cope with their diseases and rigors of treatment and clinical trials. Further research is needed to identify CAM therapies that may be used as adjunct treatments during clinical trials. PMID- 11059504 TI - A pilot evaluation of the anti-inflammatory activity of Culcasia scandens, a traditional antirheumatic agent. AB - OBJECTIVE: To screen the anti-inflammatory potentials of a popular traditional antirheumatic herb, Culcasia scandens. DESIGN: The leaves of Culcasia scandens were collected, identified, dried, and reduced to coarse powder and extracted with methanol. The methanol extract was fractionated into seven fractions. The fractions were subjected to phytochemical analysis to identify the biologically active constituents. The anti-inflammatory activity of the crude (methanol) extract (CE) and the fractions were determined in rats. The crude extract was also subjected to acute toxicity tests. RESULTS: The extract was partitioned into seven fractions (F1-F7) using preparative thin layer chromatography. Fraction F1 and the methanol-insoluble fraction (F9) did not exhibit any anti-inflammatory activity. The other fractions showed anti-inflammatory activity in the following order F6> F2> F5> F4> F8> F3> F7> CE. The anti-inflammatory potency of F2 and F6 at the dose tested were greater than that of aspirin (100 mg/kg). Phytochemical analysis of the extracts revealed the presence of reducing sugars, carbohydrates, alkaloids, glycosides, saponins, tannins, flavonoids and an unsaturated lactone ring of steroids. The median lethal dose (LD50) of the crude extract in the mice was greater than 5 g/kg. The results indicate that the leaves of C. scandens have good potential as an anti-inflammatory agent. PMID- 11059505 TI - Use of herbal therapies by adults seen in an ambulatory care research setting: an exploratory survey. AB - OBJECTIVE: To identify and characterize patterns of use of herbal products among patients participating in selected research clinics. DESIGN: Survey of three National Institutes of Health (NIH) ambulatory care research clinics. SUBJECTS: Convenience sample of 490 adult patients (168 male, 322 female) attending rheumatology, liver, and endocrinology/metabolic research clinics. RESULTS: Of the patients surveyed, 16.7%: (n = 82) reported using herbs. There were no significant sociodemographic differences between herb and nonherb users. Indications for herb use differed among the disease groups; patients in the endocrine and rheumatology clinics were taking herbs predominantly for "energy" or "wellness"; those attending the liver clinic tended to use herbal therapies as treatment for their disease. Mean and median monthly expenditure for herbal products was $30 and $10, respectively. There was a significant positive correlation between number of herbs used and use of other dietary supplements (p < 0.0001). CONCLUSIONS: One in six patients in ambulatory clinical research settings may be taking herbal products in addition to prescribed treatment. This figure is lower than in the general population, possibly because the patients may stop using herbs when participating in a research project. Although empirical evidence on the beneficial or adverse effects of herb therapy alone or in combination with drug therapies is limited, clinical researchers should be aware of the potential for confounding clinical trial results. PMID- 11059506 TI - Chiropractic through the eyes of physiotherapists, manual therapists, and osteopaths in The Netherlands. AB - OBJECTIVES: To identify current perceptions and levels of awareness of chiropractic among physiotherapists, osteopaths, and manual therapists in The Netherlands. In addition, to investigate how future communication and interprofessional collaboration between the four professions may be improved in the interests of patient care as perceived by these groups. DESIGN: Four hundred and ninety-four (494) questionnaires were distributed to 100 manual therapists, 299 physiotherapists, and 95 osteopaths across The Netherlands. Questionnaires were identical for each profession. RESULTS: An overall response rate of 48% was achieved. The majority of practitioners reported limited knowledge of chiropractic. However, chiropractic was generally perceived as a primary health care profession most suited to extramural care. Sixty-seven percent (67%) of the manual therapists regarded it as direct competition, while 45% of the osteopaths and 48% of physiotherapists considered chiropractic complementary to their professions. While the majority of osteopaths felt that statutory self-regulation should be granted to chiropractors in The Netherlands, this was not supported by the manual therapists and physiotherapists. Moreover, there was only minimal (4% 11%) support for the availability of chiropractic treatment as part of the Dutch National Health Service. Although most respondents had never had contact with a local chiropractor, all osteopaths and 50% of the manual therapists and physiotherapists considered chiropractors to be skilled practitioners. However, this was more likely to be so if they had had contact with a chiropractor in the past. Current levels of communication and cooperation were thought to be poor to nonexistent although the majority welcomed closer links, particularly in relation to the treatment of spinal complaints. CONCLUSION: Greater awareness appears to be associated with increased levels of interprofessional acceptance and respect. The professions may wish to pursue areas of broad agreement identified by their practitioners in the interests of professional development and optimal standards of care for individuals in need of musculoskeletal services. PMID- 11059507 TI - Patient risk-taking attitude and the use of complementary and alternative medical services. AB - OBJECTIVES: Study users and nonusers of complementary and alternative medicine (CAM) with respect to their attitude toward risk. SETTINGS AND SUBJECTS: National household telephone survey fielded in the United States in 1998 (n = 9,585). DEPENDENT VARIABLE: CAM use in previous year. RESULTS: Patients of CAM practitioners consider themselves more likely to take risks than the average person (odds ratio [OR] 2.47, 95 confidence interval [CI] [1.91, 3.19] compared to the general population). Risk attitude is as strong (or even stronger) a predictor of visits to CAM providers than the main sociodemographic predictors of female gender, higher education, or middle age. Individuals using only self administered CAM treatment rate themselves as being relatively more cautious (OR, 1.08; not statistically significant from the general population). CONCLUSIONS: Patients of CAM practitioners perceive themselves as risk taking, whereas patients that only rely on self-administered CAM treatment rate themselves not differently from the general population. This major difference within the group of CAM users and compared to the general population has not been studied before. PMID- 11059508 TI - Role of herbal compounds (PC-SPES) in hormone-refractory prostate cancer: two case reports. AB - PURPOSE: Herbal therapies are unconventional treatments that have been used for several different diseases. PC-SPES is an herbal mixture, composed of eight different herbs (chrysanthemum, isatis, licorice, Ganoderma lucidum, Panax pseudo ginseng, Rabdosia rubescens, saw palmetto, and scutellaria), which has been used as an alternative in the treatment of prostate cancer. We report two cases of hormone-refractory prostate cancer patients, who showed a favorable response to therapy with this herbal combination, controlling the progression of the disease. METHODS: We report two cases of biopsy proven prostate cancer patients with metastatic disease, treated with total androgen blockade, progressing to an androgen-independent status. These patients were offered traditional therapies for hormone-resistant prostate cancer, and they chose to take PC-SPES. The follow up as well as their evolution are described. RESULTS: PC-SPES extract decreased the prostate-specific antigen (PSA) value for both patients from an initial value of 100 and 386 ng/mL to 24 and 114 ng/mL after 1 year and 4 months, respectively, remaining stable until now. No gynecomastia or hot flashes were observed in these patients and the treatment was well tolerated. CONCLUSION: PC-SPES has shown a strong estrogenic in vitro and in vivo activity as an alternative tool in the management of prostate cancer patients. These cases suggest that PC-SPES might have some potential activity against hormone-independent prostate cancers. PMID- 11059509 TI - Traditional medicine and HIV/AIDS in Africa: a report from the International Conference on Medicinal Plants, Traditional Medicine and Local Communities in Africa (a parallel session to the Fifth Conference of the Parties to the Convention on Biological Diversity, Nairobi, Kenya, May 16-19, 2000). PMID- 11059510 TI - Association of school absence with air pollution in areas around arterial roads. AB - Association of school absence with air pollution from suspended particulate matter (SPM) and nitrogen dioxide was analyzed in areas around arterial roads for five years, from 1993 to 1997. The prevalence of absence was calculated using the data for school absence in two schools around arterial roads, one of which is near a crossing (Sch.A), and the other is adjacent to an arterial road (Sch.B). Although the results from annual correlation analyses did not indicate common findings for five years or in the two schools, the prevalence of absence correlated positively with SPM, nitrogen dioxide, or relative humidity, and negatively with atmospheric temperature. As the results from multiple regression analyses, atmospheric temperature in Sch.A was adopted as the optimum explanatory variable, whereas SPM and relative humidity were considered in Sch.B. Odds ratios for the prevalence of absence to SPM were elevated and were significant in Sch.B, when using a quintile method. The other odds ratios for the air pollutants were not significant, but exceeded 1. When the data were classified by day of the week, significant associations of the prevalence of absence were observed with atmospheric temperature in Sch.A and with SPM in Sch.B. The slope of the regressive equations by day of the week became steeper with the day in Sch.B. SPM was weakly associated with the prevalence of absence in Sch.A and was closely associated in Sch.B according to the optimum variables selected from the multiple regression analyses by day of the week. PMID- 11059511 TI - The risk for second primaries in gastric cancer patients: adjuvant therapy and habitual smoking and drinking. AB - STUDY PURPOSE: To examine whether adjuvant therapy for gastric cancer increases the risk for second primaries, and whether smoking and drinking increase the risk. SUBJECTS AND METHODS: 1,631 patients who were newly diagnosed with gastric cancer and underwent curative operation at Osaka Medical Center for Cancer and Cardiovascular Diseases during 1978-92. Incidence of second primaries were examined through linkage to Osaka Cancer Registry as of the end of December 1995. Observed number of second primaries (O) was compared with the expected according to the incidence in general population (E). Proportional hazards model was used to estimate hazard rate ratio (HR) for second primaries. RESULTS: 149 second primaries were observed. Seventeen cases, detected within 2 months after diagnosis of gastric cancer, were excluded. O/E ratio was 1.13 for adjuvant chemotherapy, 0.93 for immuno-chemotherapy, and 0.78 for immunotherapy, while 1.14 for operation only (not significant). Age-, sex-, and stage-adjusted HR was 1.02 (95% C.I. 0.69-1.50) for chemotherapy, 0.80 (0.41-1.57) for immuno chemotherapy, and 0.60 (0.08-4.34) for immunotherapy, as compared with the risk for operation only. Among males, HR for current smokers vs. never smokers was 1.82 (1.02-3.26). CONCLUSIONS: Adjuvant therapy for gastric cancer did not significantly increase the risk for second primaries, while smoking elevated it. PMID- 11059512 TI - Household size related to prevalence of smoking in women in Japan. AB - We investigated the relationship between the prevalence of smoking in females and various social factors, such as household size. Ten thousand and sixty-nine subjects over 20 years of age were randomly selected from the general population of Mie Prefecture. The results showed that the habit of smoking was significantly associated with household size in women (P< 0.01), but not in men. Regarding household size, current smoking rate of women aged 20-59 who live in three generation household was lower than those who live in others. Therefore, the steady replacement of the traditional three-generation household by smaller households in Japan may lead to an increase in the number of young women who smoke. PMID- 11059513 TI - Green tea consumption and chronic atrophic gastritis: a cross-sectional study in a green tea production village. AB - Chronic atrophic gastritis (CAG) is well known as a precancerous lesion of the stomach, and Helicobacter pylori (H. pylori) infection increases the risk of CAG. While recent studies have reported that green tea consumption decreases the risk of gastric cancer, there has been no study analyzing the relationship between green tea consumption and the both risks H. pylori infection and CAG. We conducted a cross-sectional study on 636 subjects living in a farming village in Japan to examine the relationship among green tea consumption, H. pylori infection, and CAG. Smoking, alcohol drinking, consumption of four beverages, including green tea, and of five foods were investigated as lifestyle factors that may affect H. pylori infection and CAG. The measurement of H. pylori-IgG antibodies was used to define H. pylori infection, and serum pepsinogens were used to define of CAG. The unconditional logistic regression model was used for analyzing each odds ratio (OR). H. pylori infection was positively associated with the risk of CAG (OR = 3.73; 95% confidence interval [CI], 2.59-5.36). High green tea consumption (more than 10 cups per day) was negatively associated with the risk of CAG, even after adjustment for H. pylori infection and lifestyle factors associated with green tea consumption (OR = 0.63; 95% CI, 0.43-0.93). These results support the hypothesis that high green tea consumption prevents CAG. PMID- 11059514 TI - Relationship between suicide and holidays. AB - Employing the vital statistics from January 1, 1979 to December 31, 1994 in Hokkaido Prefecture, Japan, we investigated the relationship between suicide (ICD 9 code, El15) and days of the week. In one day 1.97 males and 0.98 females committed suicide on average. On Saturdays the number of suicides per day was the smallest. On Mondays it was the highest. When the days were classified into (1) a holiday, (2) the day before a holiday, (3) the day after a holiday (4) the day both before and after a holiday and (5) others, the rate of suicide was the lowest in the days before a holiday. In the days after a holiday, however, the rate was the highest. The same tendency was also noted when the subjects were classified into several subgroups from the viewpoint of ages, seasons or calendar years. The time relation of holidays seemed to have something to do with the intention to commit suicide. PMID- 11059515 TI - Repeatability of the questionnaire for the aging level indices. AB - The purpose of the study is to confirm repeatability of the series of self administrating questions, which can be applied in large community populations. In 1994 and 1995, two surveys were conducted for the residents at Minamikawachi, Japan with a same questionnaire. The number of respondents for the both surveys was 887. Kappa statistics of all items of the series of questions for aging level indices were significantly high. Kappa statistics were over 0.4, and repeatability is good or excellent in 28 items of all the 45 items. The statistics of the items for medical treatments of chronic diseases were high, in such as diabetes (0.846) and hypertension (0.604). For activity of daily livings, such as shopping (0.619), kappa statistics were also high, but that of eating (0.162) was low. The statistics of subjective symptoms were moderate, however, that of impairment of hearing (0.672) was high, and that of decline of interest in opposite sex (0.256) were low. On the other hand, kappa statistics of acute diseases were low, in such as bone fracture (0.073). Correlation coefficients of the comprehensive aging level indices are around 0.6 among the people of 65 years old or older. In conclusion, the repeatability of the questions and indices were good, and they are appropriate to apply to communities. PMID- 11059516 TI - Importance of sex and age factor in assessing family history of stroke. AB - Incidence of stroke differs between men and women and it increases nearly exponentially with age. Therefore, assessment of family history of stroke disregarding sex and age of family members results in bias or misclassification. In this study the effects of sex and age on the positivity of the past history were analyzed numerically. Sex- and age-specific proportion of a positive history of stroke among 24,007 family members was obtained from a questionnaire survey of 2,316 high school students. By analyzing the sex- and age-specific proportion with the logistic regression model odds ratios resulting from sex and age difference were estimated. The odds ratio for sex difference was 2.458 (95% confidence interval: 2.067-2.924) and odds ratio for age difference was 1.064 (95% confidence interval: 1.058-1.070). This indicated that a positive history of stroke was 2.458 times higher in male members than in female members of the same age and that a positive history increased by (1.064)y, where y was age difference in years. Potential bias or misclassification resulting from disregarding sex and age can be substantial. Some measures to control for sex and age of family members are required in assessing the family history. PMID- 11059517 TI - Maternal active and passive smoking and fetal growth: A prospective study in Nagoya, Japan. AB - In order to examine the effect of maternal active and passive smoking on fetal growth, we carried out a population-based cohort study. A self-administered questionnaire was distributed to 15,207 women who notified their pregnancy from April, 1989 to March, 1991. A total of 7,411 mother-singleton infant pairs were analyzed in this study. Paternal smoking status and maternal hours exposed to environmental tobacco smoke (ETS) were used as indicators of passive smoking. Infants born to active smoking mothers were 96 g lighter, on an average, at birth than those born to non-smokers, and the relative risk for intrauterine growth retardation was 1.79 (95% confidence interval (CI) = 1.05-3.04) among active smoking mothers. Infants with smoking fathers weighted 11 g lighter, on an average, than those with non-smoking fathers, and mean birth weight of infants was reduced by 19 g among mothers exposed to ETS. The relative risk for intrauterine growth retardation in non-smoking pregnants with a smoking husband and those exposed to ETS was 0.95 (95% CI = 0.72-1.26) and 0.95 (95% CI = 0.71 1.26), respectively. Our findings indicated an adverse effect of maternal active smoking on fetal growth in Japanese pregnant population, but with small influence of maternal passive smoking. PMID- 11059518 TI - Behaviors and attitudes towards smoking among the nurses in Japan. AB - The study was carried to investigate on the actual conditions related to smoking of the nurses working in all medical institutions under a regional medical association in Mie Prefecture (regional medical institutions). Results obtained were as follows: smoking prevalence of female nurses is considered to be equal to that of the general female population in Japan. About 35% of the nurses with the smoking habit had an opinion to quit it, and about 45% of them practiced it seriously. Nearly 80% nurses favored restriction. In the way of thinking related to smoking, more than 90% of the nurses answered that women should not smoke for the health of the fetuses and infants, while only about 30% of them agreed to stop smoking working as members of the medical staff. The survey suggests that anti-smoking program is necessary to develop for smoking nurses working at medical facilities. PMID- 11059519 TI - Genetic polymorphism of enzymes involved in xenobiotic metabolism and the risk of colorectal cancer. AB - Environmental factors such as smoking cigarette, diets and alcohol may interact with genetic factors, which put one individual at a greater or lesser risk of a particular cancer than another. Advances in molecular biology have allowed many allelic variants of several drug metabolizing enzymes so that individuals with the susceptible genotypes can be determined easily. Many pieces of research have focused on the relationship between the distribution of polymorphic variants of different forms of the metabolic enzymes and colorectal cancer susceptibility because of importance roles of the metabolic enzymes in the activation of many procarcinogens or chemicals. In this respect five groups of the metabolic enzymes, cytochrome P450 (CYP) 1A1/CYP1A2, glutathione S-transferases (GSTs), N acetyltransferases (NATs), aldehyde dehydrogenase 2 (ALDH2) and methylenetetrahydrofolate reductase (MTHFR), have been discussed here. A positive association between development of colorectal cancer and the mutant homozygous genotype in Msp1 polymorphism of CYP1A1 gene has been reported in Japanese in Hawaii. The relation between genetic polymorphisms in GSTs and cancer risk has also taken an interest. At least nine studies have demonstrated the relation between the GST polymorphisms and colorectal cancer. Two of these studies suggested an increased risk of approximately 2-fold among those with the GSTM1 null genotype, while others found no risk increase. None of these studies examined the combined effect of CYP1A1 and GST polymorphisms. Either NAT2 or CYP1A2 alone have been slightly associated with colorectal cancer. When CYP1A2 and NAT2 phenotype were combined, a significant increased risk (odds ratio of 2.8) was seen among well done meat consumers with the rapid-rapid phenotype. Two published studies have found that the risk of colorectal cancer can be enhanced (2-3 fold) in alcohol drinkers with heterozygous genotype of ALDH2 in two Japanese populations recently. Findings from three published studies suggested that the mutant genotype of MTHFR inversely slightly associated with colorectal cancer. Although some of genetic polymorphisms discussed here have not shown statistically significant increase/decrease in risk, individuals with differing genotypes may have different susceptibilities to colorectal cancer, based on environmental factors. Further studies are needed to identify risk groups more specific and to determine factors of importance in colorectal cancer development. PMID- 11059520 TI - Inhaled glucocorticoid therapy in infants at risk for neonatal chronic lung disease. AB - The primary impetus for the study of inhaled glucocorticoid therapy in the treatment and prevention of neonatal chronic lung disease (CLD) was to achieve effective anti-inflammatory therapy with few adverse effects. Initial reports of inhaled glucocorticoid therapy in infants with established CLD suggest modest improvement in neonatal respiratory outcomes. Recent randomized trials also indicate that inhaled glucocorticoid therapy may provide some benefit, but have not demonstrated a reduction in CLD. Some studies suggest that the pulmonary response to systemic glucocorticoid may be greater and faster than response to inhaled glucocorticoid therapy. Few adverse effects have been noted with inhaled glucocorticoid therapy. One limitation of studies of inhaled glucocorticoid therapy is the uncertainty of the dose delivered and deposited in peripheral airways and regions of the lungs. Experience with and systematic study of inhaled glucocorticoid therapy is still in its early stages. The role of inhaled glucocorticoid therapy in the treatment and prevention of CLD is evolving. Advances in delivery devices and new developments of drug formulations should improve aerosol delivery and deposition in infants. Given the clinical dilemma of systemic glucocorticoid therapy and potential benefits demonstrated by recent trials of inhaled glucocorticoid therapy, further study of inhaled glucocorticoid therapy for CLD is warranted. PMID- 11059521 TI - Primary schoolteachers' knowledge of asthma: the impact of pharmacist intervention. AB - The Newcastle Asthma Knowledge questionnaire was used to determine primary schoolteachers' knowledge of asthma. The mean score achieved (20.71 +/- 3.20), was representative of a reasonable degree of knowledge. However, knowledge of symptoms associated with asthma and of medications used to manage the condition was poor. Teachers who had contact with an asthmatic individual or who had received previous training on asthma had a significantly greater knowledge of asthma. An asthma training session, delivered by a pharmacist, was found to significantly improve primary schoolteachers' knowledge of the condition, however, the extent of the improvement was low. PMID- 11059522 TI - A clinical trial of the Buteyko Breathing Technique in asthma as taught by a video. AB - The Buteyko Breathing Technique (BBT) is promoted as a drug-free asthma therapy. It is based on the premise that raising blood PaCO2 through hypoventilation can treat asthma. Our study was designed to examine whether the Buteyko Breathing Technique, as taught by a video, is an efficacious asthma therapy. Thirty-six adult subjects with mild to moderate asthma were randomized to receive either a BBT or placebo video to watch at home twice per day for 4 weeks. Asthma-related quality of life, peak expiratory flow (PEF), symptoms, and asthma medication intake were assessed both before and after intervention. Our results demonstrated a significant improvement in quality of life among those assigned to the BBT compared with placebo (p = 0.043), as well as a significant reduction in inhaled bronchodilator intake (p = 0.008). We conclude that the BBT may be effective in improving the quality of life and reducing the intake of inhaled reliever medication in patients with asthma. These results warrant further investigation. PMID- 11059523 TI - Children's illness drawings and asthma symptom awareness. AB - This study examines the relationship between children's abilities to perceive their symptoms of asthma via several previously researched subjective and objective procedures compared with their performance on a standardized children's drawing task and scale criteria. Results indicated that girls verbalized significantly more emotions about their drawings and were better able to detect airflow changes in their small airways than boys. The Gabriels Asthma Perception Drawing Scales (GAPDS) is a promising clinical tool for assessing children's perceptions and emotions about asthma via nonverbal methods. Varying methods of measuring asthma symptom awareness are not highly correlated; thus, more than one methodology is appropriate for use with children. PMID- 11059524 TI - Breastfeeding and asthma among Brazilian children. AB - We examined the association of breastfeeding and the presence of chronic respiratory symptoms among 5,182 Brazilian schoolchildren 7-14 years of age who were participants in the International Study on Asthma and Allergies in Childhood (ISAAC). The prevalence of medically diagnosed asthma and current wheeze were respectively 4.6% (95% confidence interval [CI] 4.0%-5.2%) and 11.9% (95% CI 11.0%-12.8%). Ninety percent of the mothers in our study population had breastfed their child. After adjusting for potential confounding factors, we found that children who had not been breastfed were more likely to have a medical diagnosis of asthma (odds ration [OR] = 1 .51, 95% CI 1.00-2.51), experience current wheeze (OR = 1.29, 95% CI 0.96-1.74), and wheeze after exercise (OR = 1.51, 95% CI 1.01 2.27) than children who had been breastfed for more than 6 months. This effect was only present among children with no family history of asthma (OR = 1.54, 95% CI 0.90-2.42 for medical diagnosis of asthma; OR = 1.27, 95% CI 0.93-1.75 for current wheezing; and OR = 1.74, 95% CI 1.12-2.6 for wheeze after exercise). We conclude that the low prevalence of asthma and wheeze observed in our population may be partly related to the high level of breastfeeding. PMID- 11059525 TI - Effect of a brief educational intervention on medical students' use of asthma devices. AB - Several studies have shown that a significant percentage of housestaff and attending physicians are deficient in both skill and knowledge of the metered dose inhaler (MDI). There are no studies involving medical students, or any including the peak flow meter (PFM). The setting was a large health science center with investigators in private conference rooms with individual medical students. Twenty-two medical students in the last semester before graduation were scored in the use of these devices pre-education and post-education (instruction included both discussion and demonstration). Results revealed a lack of skill initially, followed by dramatic improvement after the intervention. The total number of correct steps for each device (MDI with spacer and PFM) improved significantly (p < 0.0001). This group of medical students was deficient in the use of common asthma devices. A short educational intervention was effective in improving skill. PMID- 11059526 TI - Obstetric complications and asthma in childhood. AB - Studies have shown that perinatal factors are associated with childhood asthma. The current analyses examined the association between obstetric complications and risk of asthma at the age of 7 years using a prospectively population-based birth cohort in northern Finland. Results indicated that obstetric complications were associated with a higher risk of asthma among children. Those children who were administered special procedures at birth, i.e., cesarean section, vacuum extraction, and other procedures, including use of forceps, manual auxiliary, and extraction breech, had an adjusted odds ratio (OR) for asthma of 1.38 (95% confidence interval [CI] 1.00-1.92), 1.32 (95% CI 0.80-2.19), and 2.14 (95% CI 1.06-4.33), respectively, as compared to children who were delivered normally. Children who had a lower Apgar score at the first and the fifth minute after birth also had a higher risk as compared to those who had an Apgar score of 9-10. The results encourage further evaluation of the association between obstetric complications and risk of asthma among children in other populations, and further exploration of possible mechanisms underlying the association. PMID- 11059527 TI - Prevalence of nocturnal asthma in a general population sample: determinants and effect of aging. AB - Nocturnal asthma (NA) is important because of clinical and prognostic implications. Previous data on prevalence may be overestimated, because they are derived from selected series. Observations on monitoring of peak expiratory flow in elderly asthmatics suggested that prevalence of NA may increase with age. This study was designed to estimate the prevalence of NA-related symptoms in a sample drawn from a general population and evaluate the role of aging. Subjects (1,100, mean age 41.9, SD 22.8 years) were randomly selected from the lists of seven general practitioners. A questionnaire on nighttime and morning NA-associated symptoms was used and frequency of occurrence was rated as never, sometimes (less than once a week), and often (once a week or more). In the overall sample, symptoms were experienced "sometimes" by 2.3%-4.9% of subjects, whereas the response "often" was given by 0.9%-1.6% of subjects. Among subjects with a diagnosis of asthma, symptoms occurred sometimes in 16.7%-23.7% and often in 5% 15%. Symptoms reported the morning after were significantly more frequent among patients aged 65 years and older (p < 0.005), whereas the difference for nighttime symptoms was not statistically significant in different age groups, confirming an age-related blunted sensitivity. Logistic regression analysis shows that a diagnosis of asthma is the most important correlate of symptoms, with odds ratio (OR) up to 14.78 for cough; advanced age also proved to be an independent risk factor (OR 3.35-4.97). In conclusion, although the prevalence of NA was previously overestimated, our results indicate its importance, particularly among elderly patients who are exposed to a prominent risk of underdiagnosis and undertreatment. PMID- 11059528 TI - Asthma, inhaled corticosteroid use, and bone mass in prepubertal children. AB - The aim of this cross-sectional study was to describe the role of asthma, asthma severity, and medication usage in bone mineralization of prepubertal children. Asthma severity, medication usage, and physical activity were assessed by questionnaire and objective measures in 330 children. Bone densitometry and body composition were measured by dual-energy x-ray absorptiometry. Asthma ever was reported by 110 subjects (33%). A diagnosis of asthma was not associated with any deficit in bone mass, whereas usage of inhaled corticosteroids (ICS) in the last year (but not past use) was associated with deficits in bone in the total body (only after adjustment for confounders), particularly for doses of > or =400 microg/day. These observations support current recommendations with regard to ICS usage in children, but require confirmation in longitudinal studies. PMID- 11059529 TI - Poor perception of airway obstruction in children with asthma. AB - The aims of this study were to evaluate children's perception of asthma symptoms and to determine a clinically useful method for identifying poor patient perception of airway obstruction. Three methods were used to analyze the relationships among indices of lung function and perception of breathlessness in 35 children. Approximately half the children in our sample did not perceive either airway obstruction or bronchodilation. We propose that <20% improvement in visual analog scale scores post-bronchodilation may provide a simple index for identifying patients with poor perception of airway obstruction, who may be at risk for fatal or near-fatal asthma. PMID- 11059530 TI - Patients' perceptions of asthma control and impact on attitudes and self management. AB - The objective of this work was to describe the incidence of chronic breathing problems, particularly asthma-related breathing problems, in a cross-section of the United States population and to assess the perceived impact of these problems on daily living. An initial screening survey was used in a nationwide panel of 30,000 households; an in-depth follow-up questionnaire was sent to a random sample of respondents who reported a breathing problem. A sample (n = 2,685) of respondents who reported persistent cough, shortness of breath, or wheezing within the previous 2 years were sent a newly developed questionnaire. A subsample (n = 723) of respondents reported a primary diagnosis of asthma and of these, 59.4% were female and 90.2% were white. Their mean age was 37.4 years. Respondents (n = 723) characterized their level of perceived asthma control as completely controlled, well controlled, somewhat controlled, or poorly/not controlled. At least 31% of those with a perception of some control and 59% of those with a perception of poor/no control reported their breathing problems had increased in the last year. Those who perceived their asthma-related breathing problems as poorly controlled reported significantly greater symptom frequency, activity restriction, fears and concerns about their breathing difficulties, less helpful coping strategies, and less confidence in their doctor's ability to care for them. Responses to many of the questions indicated that the worst levels of disease control were associated with poorer quality of life and a more negative perception of the disease and its effects on daily living. A significant portion of the U.S. population appears to suffer from chronic breathing problems; this requires confirmation and further exploration to reduce the potential mortality and morbidity due to asthma in the United States. PMID- 11059531 TI - Child health in the new millennium. AB - In today's modern, industrialized and affluent countries, like Japan and Australia, better living conditions and hygiene, plentiful nutritious food and rapid advances in biology and medical technologies have helped to bring about dramatic improvements in child health. The previous heavy burdens of infections and undernutrition have been eliminated or can now be controlled or effectively treated. In these countries, child health standards are higher than ever and expectation of life at birth is much higher than in the past. Some of the technological advances that have helped bring about this transformation are immunization, antimicrobial therapy, successful treatment of childhood leukemias, transplantation of vital organs and implementation of genetic diagnosis and gene therapy. The use of genetically modified foods and the prospects for cloning of humans are areas of intense interest and controversy. However, these advances have their disadvantages (e.g. antibiotic-induced drug resistance). Urbanization has encouraged the 'westernization' of dietary patterns and the long-term 'lifestyle diseases' that can follow in adults. Accidents, violence and drug abuse are major problems in many parts of the world. Changes in attitudes to sexuality and the spread of HIV/AIDS is another major problem, especially in Africa and Asia. Environmental pollution and the degradation of agricultural lands, rivers and seas are also important. Ironically, standards of child health and the prospects for long life in countries like Japan are better than ever before, but social and environmental changes are presenting children and their carers with new and unanswered challenges as we enter the 21 st century and the new millennium. PMID- 11059532 TI - Coronary risks after high-dose gamma-globulin in children with Kawasaki disease. AB - OBJECTIVES: The goals of the present study were to develop a predictive coronary risk scoring system after intravenous gamma-globulin (IVGG) therapy of any dose for the different preparations currently used in the treatment of children with Kawasaki disease and to determine the predictive value of the system. The previously reported scoring systems were based on treatment with high-dose IVGG therapy at limited doses and were determined using investigative methods. METHODS: Four hundred and fifty-one patients were randomized into one of three groups and received either i.v. polyethylene glycol-treated human immunoglobulin at a dose of either 200 (n = 147) or 400 mg/kg per day (n = 152) or freeze-dried sulfonated human immunoglobulin at 200 mg/kg per day (n = 152) for 5 consecutive days. We documented 31 cases of coronary abnormalities (CA). Univariate and multivariate logistic regression was performed using 49 clinical variables and the resulting predictive model was validated. RESULTS: The duration of fever (odds (I day)/odds (- 5 days)= 0.158; 95% confidence interval (CI) 0.0385-0.648), hemoglobin (odds (Q1 = 10.3)/odds (Q3 = 11.6) = 3.97; 95% CI 1.92-8.20), IgG (odds (Q1 = 1,900)/odds (Q3=2,658)=2.72, 95% CI 1.18-6.25) and IgA (odds (Q1 =72)/odds (Q3= 160) = 0.415; 95% CI 0.253-0.680) levels after completion of gamma globulin infusion were independent predictors. The model is quasi-cross validated and has acceptable sensitivity and selectivity. The estimated risk and observed occurrence of CA coincide. CONCLUSIONS: Determinants of the risk of CA after IVGG therapy are a longer duration of fever, a lower IgG level, a higher IgA level and a lower hemoglobin level after IVGG infusion. This model is applicable for IVGG doses from 1 to 2 g/kg and for at least two different gamma-globulin preparations. PMID- 11059533 TI - Hemodynamic factors of thrombus formation in coronary aneurysms associated with Kawasaki disease. AB - BACKGROUND: We studied the mechanism of thrombus formation in coronary aneurysms based on rheological findings. METHODS: We studied 43 coronary aneurysms in 33 patients with Kawasaki disease (mean (+/- SD) age 6.1 +/- 4.3 years). These lesions were divided into three groups on the basis of maximum diameter: (i) small (group S); (ii) medium sized (group M); and (iii) large (group L) aneurysms. Using a Doppler flow guidewire and a pressure-monitoring guidewire, we measured coronary flow velocity and perfusion pressure inside aneurysms and in adjacent normal-looking vessels. We calculated the average peak velocity (APV) index, the mean coronary perfusion pressure (P) index and shear index. RESULTS: The APV index and shear index decreased significantly (p < 0.005) as the aneurysm size increased (APV index in groups S, M and L was 0.893 +/- 0.149, 0.573 +/- 0.242 and 0.128 +/- 0.131, respectively; shear index in groups S, M and L was 0.750 +/- 0.149, 0.328 +/- 0.153 and 0.020 +/- 0.028, respectively). However, coronary perfusion pressure showed no relationship to aneurysm size and was not significantly different from that in adjacent normal-looking vessels. CONCLUSIONS: The results of the present study suggest that the stagnation of flow and the reduction of shear stress in coronary aneurysms could initiate thrombus formation. PMID- 11059534 TI - Effects of heart rate and right ventricular pressure on right coronary arterial flow and its systolic versus diastolic distribution in a variety of congenital heart diseases in children. AB - BACKGROUND AND METHODS: In order to elucidate the underlying adjusting mechanism of human right coronary arterial (RCA) flow to increased right ventricular pressure (RVP) in children, we recorded RCA flow velocity in 24 pediatric cardiac patients at the orifice of its main trunk at the time of heart catheterization using the Doppler guidewire. RESULTS: The ratio of diastolic flow (DF)/total flow (TF), or the proportion of the DF time integral over a total of one cardiac cycle, had a negative correlation with heart rate (HR; r = -0.58, n = 11) in children with normal right ventricular systolic pressure (RVSP; RVSP < 35 mmHg). In contrast, the DF/TF ratio had a good correlation (r = 0.88, n = 24) with RVSP in all patients under study. The ratio of diastolic area (DA)/total area (TA), defined as the ratio of an area encircled by the aortic pressure curve above and the RVP curve below for diastole, over a total of one cardiac cycle, representing the overall effect of both HR and transcoronary pressure difference, also correlated well (r = 0.89, n = 24) with DF/TF. Total volume flow of the RCA also increased (r= 0.76, n = 24) with increases in RVSP, first by an increase in flow velocity through the RCA, during both systole and diastole, then by widening of the RCA lumen at very high pressures. These changes were initially more dependent on diastole with increasing RVSP because: (i) of a more marked augmentation of flow velocity in diastole compared with systole; and then (ii) of a significant decrease in flow velocity in systole at very high pressures. CONCLUSIONS: We clarify how the RCA manages to increase flow through it at different HR as a function of chronic RVP overload in pediatric cardiac patients. PMID- 11059535 TI - Celiac disease in Turkish children: presentation of 104 cases. AB - BACKGROUND: Celiac disease is characterized by life-long gluten intolerance. Clinical features of patients with celiac disease are variable. In the present study, clinical, laboratory and histologic features of 104 patients with celiac disease were evaluated. METHODS: Intestinal biopsy and serum antigliadin and anti endomysium antibodies were used for the diagnosis of patients. Mucosal lesions were classified according to the criteria of Marsh. Antigliadin antibodies were measured with a commercial enzyme-linked immunosorbent assay. Anti-endomysium antibodies were analyzed by indirect immunofluorescence with the use of a section of monkey esophagus. Routine hematological and biochemical analyses and measurement of immunoglobulin levels were undertaken. RESULTS: The mean (+/- SD) patient age was 5.9 +/- 4.1 years (range 10 months-16 years) and the most common symptom was diarrhea (81.7%) followed by abdominal distention, weight loss, anorexia and failure to thrive. Abdominal distention (60.6%), short stature (45.2%) and iron-deficiency anemia were the most common findings. High serum alanine aminotransferase levels were found in 38.3% of patients and these levels became normal after adoption of a gluten-free diet in all but two patients with cirrhosis. Immunoglobulin A, IgG antigliadin antibodies and IgA anti-endomysium antibodies were found in 76, 94 and 90% of patients, respectively. Biopsy of the small intestine revealed that 95, two and seven patients had type 3, type 2, and type 1 lesions, respectively, according to the Marsh classification. There was no statistically significant difference between patients and control groups with regard to antinuclear antibody, antismooth muscle antibody, anti-DNA and anticardiolipin antibodies (IgG and IgM). CONCLUSIONS: Although classical celiac disease was seen in most patients in the present study, clinical variability of the condition should be kept in mind. In particular, patients with uncommon findings, such as short stature, cryptogenic liver disease and iron-deficiency anemia, should also be screened for celiac disease. PMID- 11059536 TI - Six novel mutations of the fibrillin-1 gene in Korean patients with Marfan syndrome. AB - BACKGROUND: Mutations in the FBN1 gene, encoding fibrillin-1, result in Marfan syndrome (MFS). According to previous reports, the mutations in FBN1 share certain characteristics in each family with variable penetrance and overlapping symptoms, even in the same genotype. In the present study, we report six novel mutations and evaluate the clinical significance of these nucleotide changes. METHODS: To screen for nucleotide changes in all 65 exons of the FBN1 gene in 38 unrelated Korean patients, we performed polymerase chain reaction, single-strand conformational polymorphism (SSCP) and sequencing for the shift of the band in SSCP. RESULTS: We identified six mutations: a 2253 del 7 b.p., N1043S, C1254S, L1421F, C1895R and S2662P. CONCLUSIONS: These results suggest that many different mutations are responsible for MFS in the Korean population. PMID- 11059537 TI - Case-control study of perinatal factors and hepatoblastoma in children with an extremely low birthweight. AB - BACKGROUND: There is a significant association between hepatoblastoma and low birthweight. A case-control study was conducted to reveal risk factors for hepatoblastoma in children of extremely low birthweight (< 1,000 g). METHODS: Prenatal and postnatal histories, including parental histories, of 12 hepatoblastoma cases and 75 birthweight-matched controls were compared. RESULTS: The gestational age of the hepatoblastoma cases (23-32 weeks: median 25 weeks), tended to be lower than that of the controls (23-36 weeks; median, 27 weeks; P = 0.072). The time for an infant's bodyweight to return to the same level as the birthweight also tended to be longer in hepatoblastoma cases than in controls (P = 0.055). All hepatoblastoma cases received oxygen therapy for a period of 4-508 days (median 114 days), which was significantly longer than the 0-366 days (median 62 days) in the controls (P = 0.022). Furosemide was given to all hepatoblastoma cases and was used for a significantly longer period in these infants (6460 days; median 89 days) than in the controls (0-241 days; median 44 days P = 0.027). A univariate Cox regression demonstrated that the time taken to regain bodyweight at birth and the duration of both oxygen therapy and furosemide treatment were significantly associated with the development of hepatoblastoma (hazard ratio (HR)= 1.044, P= 0.013; HR = 1.006, P= 0.001; and HR = 1.007, P= 0.001, respectively). Although there were significant correlations between the factors, a multivariate Cox regression analysis identified the duration of oxygen therapy as a significant independent risk factor (HR = 1.006, P = 0.001). CONCLUSIONS: Oxygen therapy and furosemide treatment, along with the rate of growth, are risk factors for the development of hepatoblastoma in children of extremely low birthweight, and the duration of oxygen therapy is the most important factor in predicting the development of hepatoblastoma. Further studies are necessary to determine the real reasons for the development of hepatoblastoma and to protect children of low birthweight from the development of cancer. PMID- 11059538 TI - Urinary pyrimidine analysis in healthy newborns, infants, children, adults and patients with congenital metabolic diseases. AB - BACKGROUND: In previous reports, the reference range for urinary pyrimidine was determined on the basis of a small number of samples, with data for only a few patients being reported. In the present study, we measured urinary pyrimidine compounds in 25 healthy newborns, 33 healthy infants, 130 healthy children and 166 healthy adults. In addition, we also analyzed urinary pyrimidine compounds in various patients with abnormal pyrimidine metabolism, such as congenital pyrimidine metabolism disorders and urea cycle disorders. METHODS: We analyzed urines by high-performance liquid chromatography with column switching. Analyses were performed with both a reverse-phase column and an anion-exchange column. The columns were connected by a column switch, with all systems being controlled automatically by a computerized system controller. RESULTS: The excretion of pyrimidine compounds in patients with abnormal pyrimidine metabolism (containing heterozygotes) was out of our reference ranges. CONCLUSIONS: These results suggest that urinary pyrimidine analysis is a useful index for the diagnosis of pyrimidine metabolism disorders, urea cycle disorders and these heterozygotes. Based on this large-group analysis of healthy individuals, we were able to determine the reference ranges of urinary orotic acid, dihydrouracil and uracil for each age group. PMID- 11059539 TI - Congenital muscular torticollis: early and intensive treatment is critical. A prospective study. AB - BACKGROUND: Although the success of conservative management of congenital muscular torticollis has been well documented, relatively little is known about the determinants of this outcome, such as treatment duration and exercise frequency. The aim of the present study was to define factors related to treatment duration, to compare different frequencies and intensities of home treatment programs and their effect on the speed of recovery. METHODS: The present study was a prospective study of 45 infants (26 male, 19 female) with congenital muscular torticollis referred to the pediatric surgical outpatient during a I year period. Following a standardized initial assessment, parents were taught our intensive home treatment protocol, consisting of passive stretching exercises repeated every 3 h. RESULTS: The mean age at initial assessment was 38.6 days (range 15-120 days). Mean treatment duration was found to be 3.2 +/- 1.3 months. All patients were treated by use of our intensive protocol of passive stretching exercises (100% success). No surgery was necessary. CONCLUSIONS: In patients with early treated congenital muscular torticollis, there is no place for surgical treatment. This group of patients can be successfully treated using an intensive protocol of passive stretching exercises. In addition, this treatment protocol has a very short treatment duration compared with other standardized protocols. A successful outcome depends primarily on good cooperation with the parents, especially in developing countries. PMID- 11059540 TI - Role of infection in the development of acquired subglottic stenosis in neonates with prolonged intubation. AB - OBJECTIVE: To examine whether clinically diagnosed infection correlates with subsequent development of subglottic stenosis in intubated neonates. METHODS: Sixty-two neonatal infants intubated for more than 14 days were examined. Several risk factors for subglottic stenosis, including infection, duration of intubation, frequency of intubation, the size of the endotracheal tube etc., were evaluated by multiple logistic regression analysis. RESULTS: Infection that occurred within 14 days of intubation showed a positive correlation with subsequent subglottic stenosis. The duration of intubation, frequency of intubation and the size of the endotracheal tube did not affect the development of subglottic stenosis. The majority of infections were considered to be respiratory tract infections, including pneumonia. CONCLUSIONS: Infection occurring within 14 days of intubation is considered to be a risk factor for acquired subglottic stenosis in neonates intubated for more than 14 days. Prevention of infection within 14 days of intubation may reduce the incidence of subglottic stenosis in neonates. PMID- 11059541 TI - Investigation of the relationship between low Apgar scores and early neonatal thyroid function. AB - BACKGROUND: The aim of the present study was to assess the effects of low Apgar scores on perinatal thyroid function. METHODS: Forty full-term infants delivered by the normal spontaneous vaginal route were enrolled into the study. All babies had 1 and 5 min Apgar scores below 4. The control group consisted of 26 full-term healthy neonates. Cord blood and serum tri-iodothyronine (T3), thyroxine (T4), reverse tri-iodothyronine (rT3), free thyroxine (FT4), thyroid-stimulating hormone (TSH) and thyroid-binding globulin (TBG) determinations were performed by an enzyme immunoassay method. RESULTS: The mean values of FT4 and T4 observed in the cord blood of the study group were significantly lower compared with matched controls, whereas the mean TSH values were significantly higher. There were no differences in concentrations of T3, rT3 and TBG between the two groups. CONCLUSIONS: These results demonstrate the existence of transient hypothyroidism at birth in babies with Apgar scores below 4 delivered by the spontaneous vaginal route. PMID- 11059542 TI - Effects of bathing immediately after birth on early neonatal adaptation and morbidity: a prospective randomized comparative study. AB - OBJECTIVE: Because the risks and benefits of early bathing of newborn infants are not well established, we investigated the effects of bathing immediately after birth on rectal temperature, respiratory rate, heart rate, blood pressure, percutaneous arterial blood oxygen saturation (SpO2) and early neonatal morbidity. METHODS: The study was designed as a randomized prospective comparative study in the neonatal care unit of a university hospital. A total of 187 healthy term and near-term newborn infants, who were delivered vaginally without asphyxia, between January and December 1997 were the study subjects. We compared findings in newborns who were bathed 2-5 min after birth (n = 95) with those of a control group (n = 92) who received dry care instead. Groups were comparable with respect to gestational age, birthweight, male: female ratio, Apgar score and umbilical blood pH. Rectal temperature was measured with an electronic thermometer immediately before the intervention bathing or dry care and at 30 min and 1, 2, 3, 8 and 12 h after birth. Heart rate, respiratory rate, systolic and diastolic blood pressure and SpO2 were measured at 1, 2, 8 and 12 h after birth. The incidence of early neonatal morbidity, including hyperbilirubinemia and gastrointestinal and respiratory problems, was also compared. RESULTS: Rectal temperature changed over time postnatally in both groups (P < 0.0001, ANOVA) and there was a significant difference in rectal temperature between groups (P< 0.0001, ANOVA). Mean (+/- SEM) rectal temperature at 30 min after birth (i.e. approximately within 20 min after intervention) was significantly higher in the bathed group than in the control (dry care) group (37.30 +/- 0.06 is 37.00 +/- 0.05 degrees C, respectively; P = 0.000022). Respiratory rate, heart rate, blood pressure and the ratio of the number of infants with SpO2 90-94% and 95-100% did not differ significantly between the two groups. The incidence of early neonatal morbidity, including vomiting, acute gastric mucosal lesion, polycythemia, need for tube feeding, phototherapy and oxygen therapy, also did not differ between the two groups. CONCLUSIONS: Early bathing, minutes after birth, did not appear to adversely affect the adaptation of healthy full-term and near-term newborn infants. PMID- 11059543 TI - Usefulness of serum fibrinogen degradation product-E in sporadic cases of classical hemolytic uremic syndrome. AB - BACKGROUND: Hemolytic uremic syndrome (HUS) associated with Shiga toxin (Stx) producing Escherichia coli O157:H7 infection is one of the diseases causing acute renal failure in young children. Although HUS is still a serious disease in children, no reliable predictive markers for HUS nor markers of disease severity are available so far. Recently, we experienced a sporadic case of typical HUS caused by Stx-producing E. coli O157:H7 and detected, at the prodromal stage, a high level of serum fibrin/fibrinogen degradation product-E (FDP-E) fraction. METHODS: To assess the usefulness of FDP-E for the treatment of HUS in clinical practice, we retrospectively examined serum levels of FDP-E in 22 patients with bloody diarrhea with or without HUS. RESULTS: There were significantly increased levels of serum FDP-E in patients with HUS, but not in those without HUS. Furthermore, serum levels of FDP-E may correlate with disease severity in patients with HUS. CONCLUSIONS: These results suggest that serum levels of FDP-E may be a useful marker of HUS in clinical practice. PMID- 11059544 TI - Sepsis in children. AB - BACKGROUND: Sepsis remains lethal to children. At our institution, we have noted that approximately 2% of all hospitalized patients have had sepsis. In the present study, we analyzed episodes of sepsis that occurred in our ward. METHODS: Sepsis that occurred in our institution between January 1984 and December 1998 was reviewed and analyzed. RESULTS: Three hundred and sixty-six episodes of sepsis in 244 admitted patients were analyzed. Sepsis occurred in approximately 2% of all hospitalized patients. Forty-three of 244 patients were under 1 year of age. Eighty-seven percent (212/244) of cases had underlying diseases. Hematologic disorders or neoplasms were the most common underlying disease, comprising 55% of all patients (133/244). Two-hundred and fifty-one of 366 episodes of sepsis were acquired during hospitalization. We identified 409 causative agents. There were 25 polymicrobial infections (25/366; 7%). Gram-positive bacteria comprised 68% of all organisms (280/409). Staphylococcus aureus was the most common organism, comprising 18% of causing agents (75/409). Sixty-six organisms came from the insertion of a central venous catheter. Eighty-one patients experienced recurrent episodes of sepsis. In terms of complications, respiratory distress was the most common complication (36 episodes) and there were 15 episodes of shock. Thirty seven patients died of sepsis. Sepsis caused by Gram-negative bacteria showed significantly higher mortality than Gram-positive bacteria (11/43 (26%) vs 15/146 (10%); P= 0.053). CONCLUSIONS: In our institution, approximately 20% of septic patients were under 1 year of age and 90% had underlying diseases. The causative agents of sepsis affected the outcome. PMID- 11059545 TI - Septic arthritis in childhood. AB - BACKGROUND: The purpose of the present study was to determine whether there was a difference between septic arthritis (SA) combined with osteomyelitis and SA alone with regard to clinical and laboratory findings, such as symptoms on admission, age, sex, joint involvement and isolated micro-organisms, and a relationship between age and joint involvement in SA. In addition, we also aimed to determine the prognostic factors in SA. METHODS: The clinical and laboratory findings of 40 patients who were diagnosed with SA in our hospital were reviewed retrospectively. The diagnosis of SA was made according to the following criteria: immediate joint fluid aspiration (culture and Gram's stain positive, leukocyte count markedly elevated and glucose level low), blood culture positive and positive cultures from other possible sites of infection. RESULTS: Of the 40 patients, 22 were boys, 18 were girls and the male to female ratio was 1.2/1. Patient ages ranged from 6 months to 14 years (mean (+/- SD) 8.44 +/- 4.18 years). The most observed symptoms were fever (52.5%), arthralgia (50%) and joint swelling (45%). Thirty-four (85%) patients had only one joint and six patients (15%) had more than one joint involved. In total, arthritis was diagnosed in 49 joints. The joints diagnosed as having arthritis were the following: knee (n = 18), hip (n = 12), ankle (n = 12), elbow (n = 3), shoulder (n = 2), wrist (n = 1) and interphalangeal joint (n = 1). Of the 40 patients, 21 (52.5%) had SA alone and 19 (47.5%) had arthritis together with osteomyelitis. While arthritis was diagnosed in 27 joints in the group of patients with SA, it was diagnosed in 22 joints in the group of patients with SA combined with osteomyelitis; in the latter, an increase was not observed in the number of joints involved. Joint fluid culture was positive in 22 (55%) patients; the growth of Staphylococcus aureus was observed in 20 cases and Pseudomonas aeruginosa and Staphylococcus epidermidis were isolated in each patient. In contrast, in one patient, arthritis occured during meningococcal meningitis (in this patient, Gram-negative diplococci was isolated from a cerebrospinal fluid culture). Patients with SA combined with osteomyelitis and those with SA alone were compared for symptoms on admission, the history of trauma and antibiotic use, sex, age, fever, joint involvement, anemia, leukocytosis and micro-organisms isolated from joint fluid and blood; there were no significant differences for these parameters between the two groups (P > 0.05). In addition, we found that there was no relationship between age and joint involvement in SA and there was no effect of micro organisms on mortality. Three of 40 patients died; the mortality rate was 7.3%. Of the three patients who died, two had SA alone and one had SA combined with osteomyelitis. The primary disease was sepsis in these three patients; S. aureus was cultured from blood in two patients and Gram-positive cocci was observed following examination of the joint fluid in the other patient. CONCLUSIONS: We would like to emphasize that SA is mono-articular, frequently localized in the knee, hip and ankle in 85% of patients, joint fluid culture was positive in 55% of patients, bacteria was isolated from one or more cultures of blood, joint fluid, pus or bone in 70% of patients and the most common isolated micro-organism was S. aureus. In addition, it must be pointed out that children younger than 2 years of age with fever, a positive trauma history and/or abnormal joint findings should be carefully examined for SA because the rate of SA was lower (7.5%) than expected in this age group. We also found that the mortality of SA was not influenced by age, joint involvement and bacterial agents, and there was no significant difference in symptoms on admission, the history of trauma and antibiotic use, sex, age, fever, joint involvement,anemia, leukocytosis and micro organisms isolated from joint fluid and blood between patients with SA PMID- 11059546 TI - Serum C-reactive protein in the differential diagnosis of childhood meningitis. AB - BACKGROUND: Although determination of serum C-reactive protein (CRP) is considered one of the most useful tests for differentiating between bacterial and aseptic meningitis, its diagnostic accuracy in comparison with other laboratory parameters is yet to be further evaluated. METHODS: A total of 192 pediatric cases, aged between 2 months and 14 years, comprising patients with bacterial meningitis (n = 66) and aseptic meningitis (n = 126), were retrospectively analyzed on the basis of data from the initial examination. The area under the best fit binormal curve of the receiver operating characteristics (Az) for CRP was determined and compared with that for several other analytic parameters, including white blood cell count and erythrocyte sedimentation rate of peripheral blood, standard cerebrospinal fluid analysis variables and the combination test (probability of acute bacterial meningitis (pABM)) derived from Hoen's model. RESULTS: Compared with each of the other variables, the Az for serum CRP (0.97 +/ 0.02) was found to be significantly greater (P < 0.01) for all except pABM (0.99 +/- 0.01; P > 0.05). False-negative cases among the CRP test results were found to have been examined too early. CONCLUSIONS: The diagnostic accuracy of a single CRP determination was found to be equivalent to that of the most effective combination test. Patients with meningitis in whom serum CRP values are determined at least 12 h after the onset of fever and are < 2 mg/dL are far less likely to have bacterial meningitis. PMID- 11059547 TI - Parental concern towards the use of inhaled therapy in children with chronic asthma. AB - BACKGROUND: Parental attitudes towards the use of inhaled therapy in children with chronic asthma influence treatment adherence and outcome. In the present study, we evaluated the perceptions and concerns of parents of children with chronic asthma towards inhaled therapy. METHODS: A self-administered standard questionnaire was distributed to parents of children attending the Paediatric Asthma Clinic. All these children required inhaled steroids for treatment. RESULTS: One-hundred and twelve of 170 parents (66%) surveyed were concerned with inhaled therapy. The most common concern with its use was medication side effects (91%), followed by 'inhaler dependency' (86%), cost of the inhaler (34%) and difficulty in using the inhaler (15%). Parental perception that the oral route was superior to the inhaled route, preference for the oral route for asthma prophylaxis and a higher steroid dose required for prophylaxis were more likely to be associated with concerns towards inhaled therapy. More importantly, these children were also more likely to miss > 25% of their prescribed doses of inhaled steroids (46 vs 22% in the group concerned about inhaled therapy compared with the group that was not concerned, respectively; P = 0.007) and had a higher mean number of nebulization treatments in the last year (3.2 +/- 2.9 vs 1.8 +/- 1.3 in the group concerned about inhaled therapy compared with the group that was not concerned, respectively; P = 0.01). CONCLUSIONS: A significant proportion of parents whose children were on inhaled prophylaxis had concerns towards the use of inhaled therapy. Parental concern towards inhaled therapy appeared to increase the problem of non-adherence to treatment. Education for these parents will need to be addressed to improve asthma management in our patient population. PMID- 11059548 TI - Nasopharyngeal colonization with penicillin-resistant Streptococcus pneumoniae in Turkish children. AB - BACKGROUND: Streptococcus pneumoniae is one of the major infectious agents observed in children. In spite of the fact that penicillin is preferred in the treatment of infections caused by S. pneumoniae, there has been a world-wide increase in the frequency of penicillin-resistant S. pneumoniae. METHODS: One hundred and fifty sick children with a clinical diagnosis of pneumonia, meningitis, acute otitis media, acute sinusitis and septicemia or bacteremia, and 150 healthy children without any infection were examined. Streptococcus pneumoniae, which were isolated from the nasopharynx, were analyzed with respect to penicillin susceptibility using the agar dilution method. RESULTS: The S. pneumoniae carriage rate was observed to be 43.3% in the group of sick children and 30.0% in the control group (P < 0.05). The penicillin resistance of S. pneumoniae isolated from the nasopharynx was determined to be 35.4% from a total of 110 isolates, with an intermediate resistance of 32.7% and a high resistance of 2.7%. The penicillin resistance of S. pneumoniae carried in the nasopharynx was determined to be 41.5% in the group of sick children and 26.6% in the control group (P > 0.05). Resistance rates of other antibiotics were determined as follows: cefotaxime 2.7%, erythromycin 19%, clarithromycin 5.4%, tetracycline 21.8%, trimethoprim-sulfamethoxazole 4.5% and rifampin 0%. CONCLUSIONS: Penicillin resistance of S. pneumoniae has recently become a problem in Turkey. Because of this, we require new strategies to limit the spread of drug-resistant S. pneumoniae. PMID- 11059549 TI - Diagnosis and follow up in four cases of incontinentia pigmenti. PMID- 11059550 TI - Spondyloepiphyseal dysplasia tarda with progressive arthropathy: an important form of osteodysplasia in the differential diagnosis of juvenile rheumatoid arthritis. PMID- 11059551 TI - A case of achondroplasia with severe respiratory failure, profound developmental delay and hypercreatine phosphokinasemia. PMID- 11059552 TI - Asthma and gastroesophageal reflux in a girl with urticaria pigmentosa. PMID- 11059553 TI - Transcatheter coil closure of a coronary artery fistula in a 2-year-old child. PMID- 11059554 TI - Bronchial polyp in a child with endobronchial tuberculosis under fiberoptic bronchoscopic observation. PMID- 11059555 TI - A case of mesenteric panniculitis in a 4-year-old child. PMID- 11059556 TI - Campylobacter jejuni bacteremia in an immunocompetent Japanese child. PMID- 11059557 TI - Deletion of the long arm of chromosome 2 (2q22-q24.2): case report and review of the literature. PMID- 11059558 TI - A case of esophageal atresia with tracheoesophageal fistula, imperforate anus, persistent cloaca, incomplete fusion of the labium and chronic renal failure due to urethral obstruction. PMID- 11059559 TI - Sleep duration of young children is affected by nocturnal sleep onset time. PMID- 11059560 TI - Incomplete form of chronic infantile neurological cutaneous and articular syndrome. PMID- 11059561 TI - Abnormally low ratio of cholic acid to chenodeoxycholic acid due to a deficiency of 3-oxo-delta4-steroid 5beta-reductase. PMID- 11059562 TI - A case-control study of toenail selenium and cancer of the breast, colon, and prostate. AB - To study the possible role of dietary and supplementary selenium intake in the etiology of cancer, we carried out a case-control study of breast, colon, and prostate cancer in Montreal between 1989 and 1993. In this study, we were able to interview a total of 1,048 incidence cases of colon (402), breast (414) and prostate (232) cancer subjects and 688 population-based controls matched for age and gender. Of these, a total of 501 cancer cases and 202 controls produced toenail samples for their selenium concentrations, which were determined by neutron activation analysis. We found no association between toenail selenium and breast cancer (odds ratio [OR], 0.72; 95% confidence interval [CI], 0.4-1.31) or prostate cancer (OR, 1.14; 95% CI, 0.46-2.83), though we did observe a statistically significant inverse association between toenail selenium level and the risk of colon cancer for both genders combined (OR, 0.42; 95% CI, 0.19-0.93; P = .009) and for female subjects (P = .050). We also found that nonsmoker case and control subjects had higher selenium in their toenail samples. This could be due either to the nature of tobacco, which reduces selenium absorption, or to smokers' consumption of certain foods containing less selenium. Further epidemiologic studies are required to clarify the role of selenium in the etiology of certain cancers. PMID- 11059563 TI - Production of tumor necrosis factor-alpha and interferon-gamma from human peripheral blood lymphocytes by MGN-3, a modified arabinoxylan from rice bran, and its synergy with interleukin-2 in vitro. AB - Recently, we presented evidence for the role of MGN-3, an enzymatically modified arabinoxylan extracted from rice bran, in potent activation of human natural killer (NK) cell function in vivo and in vitro. In the current study, we examined the mechanism by which MGN-3 elevated NK cytotoxic activity. We did this by testing the action of MGN-3 on the levels of both tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) secretions and MGN-3 function on the expression of key cell surface receptors. Peripheral blood lymphocytes were treated with MGN-3 at concentrations of 0.1 mg/ml and 1 mg/ml, and supernatants were subjected to enzyme-linked immunosorbent assay. Results showed that MGN-3 is a potent TNF-alpha inducer. The effect was dose-dependent. MGN-3 concentration at 0.1 and 1 mg/ml increased TNF-alpha production by 22.8- and 47. 1-fold, respectively. MGN-3 also increased production of IFN-gamma but at lower levels as compared to TNF-alpha With respect to key cell surface receptors, MGN-3 increases the expression of CD69, an early activation antigen at 16 hours after treatment. Furthermore, the interleukin-2 receptor CD25 and the adhesion molecule ICAM-1 (CD54) were upregulated after treatment with MGN-3. Treating highly purified NK cells with MGN-3 also resulted in increased levels of TNF-alpha and IFN-gamma secretion in conjunction with augmentation of NK cell cytotoxic function. Furthermore, addition of MGN-3 to interleukin-2-activated NK cells resulted in a synergistic induction of TNF-alpha and IFN-gamma secretion. Overall, our data suggest that MGN-3, a novel biological response modifier, can be used as a safe alternative or as an adjuvant to the existing immunotherapeutic modalities. PMID- 11059564 TI - Inhibition of the cytochrome P-450 modulates all-trans-retinoic acid-induced differentiation and apoptosis of HL-60 cells. AB - We studied the effects of inhibition of cytochrome P-450 by proadifen (SKF525A) on the processes induced in myeloid leukemia HL-60 cells by all-trans-retinoic acid (ATRA). The parameters reflecting cell proliferation, differentiation, and apoptosis were detected by flow cytometry as the principal method at selected time intervals (24-96 hours). Changes in the expression of Bcl-2 protein were detected by Western blotting. The majority of experiments were designed as a factorial combination of the treatment and assessed for significance of the interactions. Proadifen was demonstrated synergistically (1) to potentiate the antiproliferative and differentiation effects of ATRA, and (2) to increase cell viability and prevent ATRA-induced apoptosis. Moreover, proadifen weakened ATRA induced downregulation of the Bcl-2 protein. Our results may be of practical importance because cytochrome P-450 inhibitors are used clinically in treating cancer patients. Assuming that effects on the leukemic cells in vivo would be similar, this type of combined therapy could help to achieve better results even with lower doses of ATRA. PMID- 11059565 TI - Altered expression of p53 and p27 proteins, alone or combined, as a predictor of metastatic potential in early invasive carcinoma of colon and rectum--a comparative clinicopathologic and molecular analysis. AB - To place the choice of therapy (endoscopic resection or radical surgery) in early invasive carcinoma (EIC) of colon and rectum on a more rational basis, this study sought to identify molecular predictors of metastasis. Several morphologic risk factors (histologic type, degree of tumor invasion, lymphatic and venous invasion) and expression of p53 and p27 proteins in the primary tumor were compared in 80 patients with EIC, including 12 (15%) with metastasis or recurrence (or both). Of the factors enumerated, deeper invasion of the submucosal layer, lymphatic-venous invasion, p53 overexpression, and decreased expression of p27 were correlated significantly with metastasis. The results also indicated that altered expression of p53 or p27 is independently relevant to metastasis of EIC. Analysis of these markers, together with determination of the morphologic risk factors, could complement the identification of patients with metastasis on the basis of known morphologic risk factors. Because the molecular factors can be assessed more objectively than can the morphologic parameters, they may strengthen the ability to identify EIC that has undergone, or will undergo, metastasis. PMID- 11059566 TI - Cytochrome P450 isoenzyme mRNA expression pattern in human urinary bladder malignancies and normal urothelium. AB - This study was designed to analyze the cytochrome P450 isoenzyme mRNA expression pattern of transitional cell carcinomas of the bladder (N = 19) and normal urothelium (N = 10). In addition, biopsies from normal urothelium (N = 32) taken at the time of transurethral resection of bladder cancer in eight patients from surrounding histologically normal urothelium also were characterized concerning their specific cytochrome P450 mRNA expression pattern. A total of 13 of 19 of the analyzed tumor specimens (68%) revealed expression of cytochrome P450 1B1. Cytochrome P450 4B1 and 1A1 mRNA expression were detected in 79% (15 of 19) and 53% (10 of 19) of the tumor specimens, with no correlation between tumor stage and grade of the neoplasm. Biopsies from macroscopically and histologically normal urothelium from tumor-invaded bladders also showed expression of cytochrome P450 1B1 in 75% (24 of 32), 4B1 in 62.5% (20 of 32), and 1A1 in 50% (16 of 32). Furthermore, a 75% homology concerning cytochrome P450 1B1 and 4B1 mRNA expression was observed between the bladder tumor and the biopsies from this bladder. The polymerase chain reaction analysis of normal urothelium from normal bladders that do not harbor a neoplasm revealed CYP450 mRNA expression for CYP450 1A1 in 6 of 10; 1B1 in 5 of 10; 4B1 in 6 of 10; 2D6 in 2 of 10; and 2E1 in 2 of 10. According to our data, CYP450 1B1 mRNA expression is not tumor-specific. The present findings are the first to compare CYP450 expression in bladder cancer with biopsies from the same tumor-bearing bladder, and they indicate that, from the enzymatic point of view, bladder cancer also is a panurothelial field disease present in even normal urothelium. PMID- 11059567 TI - Clinical application of NMP22 in the management of transitional cell carcinoma of the bladder. AB - The combination of a noninvasive, quantitative immunoassay, NMP22, with voided urinary cytology prior to cystoscopy was evaluated in patients with urothelial transitional cell carcinoma. Fifty-six patients with a history of transitional cell carcinoma were evaluated. Voided urine was obtained for NMP22 and cytology prior to cystoscopy. One hundred and twenty-three NMP22 assays, 124 cytologies, and 124 cystoscopies were performed. The type of anesthesia used for cystoscopic evaluation was determined by the NMP22 value in 30 patients. Cystoscopy results were considered positive on biopsy-confirmed malignancy. The reference value used for NMP22 was 10.0 U/ml. NMP22, cytology, and the combination of NMP22 and cytology were compared to cystoscopy and to pathologic grading and staging. Thirty-four recurrent transitional cell carcinoma episodes occurred; 22 were low grade (I-II), and 12 were high-grade (III-IV). Twenty-seven were stage Ta; four were T1; and three were T3b or 4. Within this group, NMP22 detected low- and high grade tumors equally, as compared to cytology, which was sensitive only to high grade tumors. Nineteen patients were NMP22-negative and underwent cystoscopy under topical anesthesia; 17 were tumor-free. Eleven patients were NMP22-positive and had anesthesia, and all had visible lesions, which were subjected to biopsy and were resected. Six lesions were tumors, five were inflammatory. Overall sensitivity of combined NMP22 and cytology was 70%; specificity was 72%; positive predictive value was 54%; and negative predictive value was 77%. An accurate assessment of the risk of a bladder cancer can be obtained with NMP22, cytology, and cystoscopy in patients with a history of bladder cancer. NMP22 values can be used to determine the level of anesthesia for cystoscopy in patients with a history of bladder cancer. PMID- 11059568 TI - Ocular effects of fenretinide, a vitamin A analog, in a chemoprevention trial of bladder cancer. AB - Fenretinide is a vitamin A derivative under investigation in cancer prevention trials. Because all available pharmacologic and toxicologic data were obtained from breast cancer patients, we measured plasma drug, metabolite, and vitamin A levels and studied their relationship with visual and ocular symptoms in a cohort formed mostly by male subjects belonging to a bladder cancer prevention trial. After 1 year, the mean plasma retinol levels (+/- standard deviation [SD]) were 168.2 +/- 75.8 ng/ml in 31 subjects treated with fenretinide and 594.5 +/- 168.4 ng/ml in 36 control subjects (P < .001). Plasma retinol levels were correlated inversely to drug and metabolite concentrations, which in turn were correlated inversely to the interval from last drug intake. The decline of plasma vitamin A levels accounted for a 41.7% cumulative incidence of diminished dark adaptability in the retinoid arm as compared to 6.8% in the control arm (odds ratio = 13.8; 95% confidence interval, 2.9-66.1). Although compliance as assessed by capsule count was high, three subjects originally assigned to the treatment group who proved to be noncompliers (8.8%, or 3 of 34) had no detectable plasma drug or metabolite levels. Our data confirm the specific pharmacologic and visual effects of fenretinide also in a male population and strengthen the importance of multiple blood measurements to monitor treatment compliance in prevention trials. PMID- 11059569 TI - Detection of minimal residual disease in patients with cancer: a review of techniques, clinical implications, and emerging therapeutic consequences. AB - The issue of minimal residual disease (MRD) manifesting itself by the presence of undetected disseminated isolated tumor cells in both tissues and hematopoietic autografts from patients with early-stage malignancies or from patients in clinical complete remission has been discussed widely during the last decade. Based on the current understanding of the pathogenesis of malignancy, disseminated tumor cells persisting after conventional oncologic treatment modalities or after reinfusion of contaminated autologous hematopoietic cells constitute the source of subsequent recurrence of disease. Accordingly, much emphasis is placed on the detection and characterization of disseminated isolated tumor cells in both basic and clinical research. This effort is aimed at a better understanding of the processes of metastasis and tumor dormancy and, ultimately, the estimation of prognosis, molecular monitoring, and the design of new therapeutic agents in oncology. In our review, we used computerized (MEDLINE, Embase) and manual searches to summarize laboratory and clinical data concerning MRD focusing on the issue of MRD in solid malignancies. We give a detailed overview of the methods used for the detection and molecular characterization of disseminated tumor cells and of the prevalence and prognostic significance of the detection of MRD in patients and hematopoietic autografts. Finally, we discuss the emerging therapeutic consequences of the detection of disseminated tumor cells, with special emphasis on the therapeutic potential of antibodies. We conclude that the detection of MRD represents a hallmark for the diagnosis, monitoring, and treatment of malignant conditions in future clinical trials. PMID- 11059570 TI - Application of boundary element method to calculation of the complex permittivity of suspensions of cells in shape of Dinfinityh symmetry. AB - A numerical method using the boundary element method was developed to calculate the complex permittivity of suspensions of particles in the shape of Dinfinityh symmetry covered with a shell phase. It was an extension of the analytical methods based on Maxwell-Wagner-Sillars' effects in suspensions of shelled ellipsoids. This method was applied to particles, which were relevant to budding yeast cells and erythrocytes, to examine the effects of the shape on frequency dependence of the permittivity and conductivity of their suspensions. Results of the calculations showed that the permittivity and conductivity at high frequencies were insensitive to the change in the shape. The change in shape affected the permittivity and conductivity at low frequencies and their frequency dependence in the intermediate frequency region. This behavior could not be imitated by the calculation using analytical methods with shelled spheroid models. PMID- 11059571 TI - Thresholds for electromagnetic field-induced hypoxia protection: evidence for a primary electric field effect. AB - We have recently reported that weak electromagnetic (EM) field exposure of chick embryos induces a response that can be used to protect against subsequent hypoxic insult. This work is continued here with an exposure response study using 20-min exposure to 60 Hz magnetic fields over a range of 2-10 microT. Once again, the biomarker used was induction of hypoxia protection. A sigmoidal response curve was found, with exposures to magnetic field strengths > or = 4 microT inducing maximum hypoxia protection (68% survival). We also attempted to determine whether the magnetic or induced electric component of the EM field was responsible for the observed protection. This was accomplished by making measurements with two different orientations of the magnetic fields (perpendicular and parallel to the major axis of the egg). Owing to the configuration of the embryo in the egg, the induced electric field at the embryo was lower when the magnetic field was parallel to the major axis even though the magnetic field strength was the same for each orientation. Exposure of the embryos to the parallel orientation resulted in a reduced protective response. An exposure-response curve generated for this orientation of the field also showed a more "drawn-out" appearance, consistent with the observed distribution of embryo positions within the egg. Our results suggest that the induced electric, not the applied magnetic field, plays a primary role in the protective effect observed in this chick embryo model. PMID- 11059572 TI - Electromagnetic field-induced protection of chick embryos against hypoxia exhibits characteristics of temporal sensing. AB - We previously studied the response of mammalian cultured cells to weak, 60 Hz electromagnetic (EM) fields. Two time constants, similar to those observed in chemotaxis, were found to govern the cellular response to the field. We concluded that a system of temporal sensing, similar to that employed in chemotaxis by motile bacteria, was operative. We termed the shorter time (approximately 0.1 s) the "sensing" time, and the longer time (approximately 10 s) the "memory" time. To investigate the possibility that temporal sensing was a general property of EM field-cell interaction, the temporal properties of another EM field-induced effect was studied. The EM field-induced protection against the effects of extreme hypoxia was examined in chick embryos. Embryos were exposed to 60 Hz magnetic fields, the amplitudes of which were regularly altered throughout the 20 min exposure. Alteration was accomplished either by turning the field off and on at regular intervals (1-50 s), or by introducing brief (10 or 100 ms), zero amplitude gaps, once each second, throughout exposure. When the field was turned on and off at 0.1 s intervals, the protective effect conferred by a constant field was lost. At progressively longer on/off intervals, protection was progressively restored, maximizing at intervals of 10-30 s. Gapping the magnetic field for 10 ms, each second of exposure conferred the same protection as that observed for an uninterrupted field, but gapping the field at 100 ms each second produced a significant reduction in protection. These data exhibit remarkable consistency with those obtained in similar temporal studies of the magnetic field induced enhancement of ornithine decarboxylase activity in L929 fibroblasts. It appears that temporal sensing is a general feature of the EM field-cell interaction. PMID- 11059573 TI - Changes in neurite outgrowth but not in cell division induced by low EMF exposure: influence of field strength and culture conditions on responses in rat PC12 pheochromocytoma cells. AB - The effects of low electromagnetic field (EMF) exposure (4.5-15.8 microT, 50 Hz AC) on neurite outgrowth and cell division in rat PC12 pheochromocytoma cells were examined. The study involved two separate experimental series in which culture conditions during exposure to the magnetic fields differed. In series 1 (14 experiments in which culture conditions were not strongly conducive to cell differentiation [15% serum]), exposure to 4.5-8.25 microT for 23 h significantly inhibited neurite outgrowth by 21.5 +/- 1.3% (by Manova, p = 0.003). In contrast, in series 2 (12 experiments in which culture conditions promoted cellular differentiation [4% serum]), exposure to 4.35-8.25 microT for 23 h significantly stimulated neurite outgrowth by 16.9 +/- 1.1% (by Manova, p = 0.009). Thus, in both series, exposure to a narrow range of low EMF has significant, but opposite effects on neurite outgrowth. Exposure to higher fields, 8.25-12.5 microT (series 1) and 8.25-15.8 microT (series 2) had no significant effect on neurite outgrowth. These data, when considered with other reports, suggest that neuronal differentiation can be altered by low level EMF exposure. While this may not be detrimental, it merits further research. At present, the reasons for the significant changes in neurite outgrowth being confined to the same narrow field strength are unclear. As stated above, culture conditions in series 2 were more conducive to cell differentiation than those in series 1. This is reflected in the lower number of cells in control samples in series 2, at the end of the 23-h incubation, than in series 1 (- 16.9 +/- 1.7%, p = 0.003). As the same numbers were plated in both series, the medium used in series 1 allows more of the PC12 cells to divide; this is consistent with some cells reverting to a non-neuronal adrenal chromaffin phenotype [L. Greene, A. Tischler. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc. Natl. Acad. Sci. U. S. A., 73 (1976) 2424-2426]. Exposure to both ranges of magnetic fields (4.35-8.25 and 8.25-15.8 microT) has no effect on cell division. Thus, there is no evidence in this study that there is a mitogenic effect arising from low EMF exposure. PMID- 11059574 TI - The effects of medium-strength electric impulses on human blood. AB - Leukocyte subsets, total leukocyte isolates or full blood samples were subjected to medium-strength square-wave electric impulses (100 V/cm field force, 5 ms duration). On the surface of the leukocytes, the expressions of several markers (CD3, CD4, CD8, CD11a, CD11b and ICAM-1) were determined in order to study the influence of pulsed ionic currents on different aspects of the cellular immune response. Large individual differences were observed among randomly chosen healthy donors, both in the initial expression rate and in the response patterns of different antigens. As a general conclusion, it can be stated that electric impulses with the above parameters activate the state of immune response alertness of human leukocytes. Changes in the activities of several enzymes in the serum in response to electric impulses were also tested in order to examine the feasibility of ex vivo electric treatment of human blood for the establishment of an antiviral and immune activated condition. Slightly elevated lactate dehydrogenase (LDH) levels point to a possibility of enhanced haemolysis, while the lack of an elevation in the membrane-bound peroxidase activity indicates the absence of haemolysis. Significant rises were detected in the serum superoxide dismutase (SOD) activities. Since most ex vivo blood manipulations are characterised by the appearance of superoxide radicals in the serum, a SOD activity enhancement is considered beneficial in these cases. A mild, but significant reduction in the blood clotting time indicates that electric treatment of human blood should be performed with special attention to thrombosis prone conditions, and adequate precautions and countermeasures should be introduced. Although wider examinations are required before this method can be fully recommended, ex vivo blood treatment with medium-strength electric impulses seems to be a promising adjuvant course for the establishment of acute immune potentiation and an antiviral state in patients undergoing dialysis treatment. PMID- 11059575 TI - Electrochemical studies of the effect of lanthanide ions on the activity of glutamate dehydrogenase. AB - Using the electrochemical method based on the "diffusion" layer concept of the convective system, the effect of lanthanide ions La3+ and Eu3+ on the activity of glutamate dehydrogenase (GLDH) has been determined. In suitable concentrations, the lanthanide ions La3+ and Eu3+ can activate the GLDH in the reductive amination of alpha-Ketoglutarate (alpha-KG). The activation mechanism would be that the lanthanide ions could interrupt the binding of NAD+ to GLDH by combining preferentially to NAD+. This mechanism was proposed by the voltammetric studies. PMID- 11059576 TI - Digression on chemical electromagnetic field effects in membrane signal transduction--cooperativity paradigm of the acetylcholine receptor. AB - There is ongoing public concern on potential hazards and risks of even small electromagnetic fields (EMFs) such as those emanating from electrical appliances. Evolution and persistence of life in the natural geofields and basic scientific experience in using technical EMFs (F) suggest that, in general, living matter is remarkably stable against external field perturbations within the technical safety limits of the EM field strengths. Besides the trivial primary effects of EMF on ionic charges and dipolar matter, it is explicitly elaborated that cellular biochemical reactivity and channel transport processes are field dependent. However, equilibrium (K) and rate (k) constants are only sensitive to F if the reaction moments deltaM are finite, as seen in the general van't Hoff relationship d ln K/dF = deltaM/RT. Indeed, it is the difference (deltaM) in the electric or magnetic moments (M), representing the difference in the field forces on the reaction partners, that determines the extent of the field-induced transitions, say from an inactive conformer of a macromolecule to an active one. If small EM fields, which locally can only cause small shifts in K and k, are to become effective for chemical reactivity, amplification is required. A widely encountered concept of chemical amplification is structural, and thus functional, cooperativity, realized in many biopolymers. The cooperation of n units of such a polymer yields a larger deltaM(n)= ndeltaM and exponentially increases the field sensitivity of the overall equilibrium constant K(n) = Kn. Using the acetylcholine receptor protein as an example for signal amplification by structural cooperativity, explicit proposals are specified for the presumed amplification of small local field effects on proteins of the classical signal transduction cascades. Electric membrane field amplification by interfacial polarization in external fields is discussed in the context of using electric field pulses to transiently permeabilize cells and tissue for the direct transfer of effector substances and genes in cancer and gene therapy. PMID- 11059577 TI - Effects of nonionic surfactants on electrochemical behavior of ubiquinone and menaquinone incorporated in a carbon paste electrode. AB - A carbon paste electrode, in which the carbon particles were coated with a thin layer of a nonionic surfactant (NIS), was constructed with a pasting liquid containing ubiquinone (UQ) or menaquinone (MQ). It has revealed that the layer acts not only as a diffusion barrier but also as a matrix for the redox reaction of quinones at electrode surface, and its effects on the electrochemical behavior of quinones depend on both the physico-chemical structure of a surfactant and the kind of quinones. Further, such a modification was applied to the preparation of an enzyme electrode in which the quinone molecule act as a redox mediator and the influences on the sensitivity of the glucose biosensor was demonstrated. PMID- 11059578 TI - Kinetics of uptake of cadmium by Chlorella marina in different media. AB - The kinetics of cadmium binding to living Chlorella marina cells during a short time (a few minutes) has been studied in seawater conditions, being also considered separately the influence of calcium and chloride. Cadmium complexes with surface groups of C. marina cells are labile in terms of differential pulse anodic stripping voltammetry in media (0.01-0.7) M NaNO3 since the complexation reaction is kinetically fast. However, in media 0.7 M NaCl, 0.7 M NaCl + 10(-2) M Ca2- and in synthetic seawater diluted 1:4 or not, the complexes are inert, since kinetics is determined by the rate of exchange with alkaline-earth metals and/or the chloride anion. PMID- 11059579 TI - Studies of mitochondrial porin incorporation parameters and voltage-gated mechanism with different black lipid membranes. AB - Our work in general aims to clarify the mechanism of what can be considered as a process of the kinetics of porin incorporation into bilayer planar membranes and to identify the parameters involved. In this paper, we report the results of systematically investigating the kinetics of porin incorporation into bilayer membranes made up of phosphatidylinositol or oxidized cholesterol using a simple and low-cost ac method. By means of a mathematical model, we provide evidence that two concurrent processes are present during the kinetics which can be interpreted as positive/negative cooperativity, and we investigate the parameters' dependence on external applied voltages. We observed a phase transition (or similar phenomenon) which seems to take place during the insertion process. The conductance measurement obtained by using data at the steady state conditions, provided indirect indications of two possible gating mechanisms. PMID- 11059580 TI - Structure of phosphatidyl serine (PS) involved in interbilayer salt bridging and hydrogen bonding. AB - For understanding the experimental results indicating salt bridging and hydrogen bonding between opposite polar surfaces in planar multibilayer structures of phosphatidyl serine (PS), (1) we carried out molecular modeling of the interacting surface layers. The interacting structures in the planar multibilayers are stabilized by salt bridge arrays of the phosphates with their counterions and by hydrogen bonds of ammonia from one polar surface with carbonyls in the opposite one. In multishell liposomes, where the distances between phosphates on the facing each other surfaces are not equal and they are bound to get out of register, the interbilayer interaction cannot extend over a large enough area to form stable structures, except if the salt bridges are strong enough to break down the curved surfaces and form planar multibilayers. PMID- 11059581 TI - Efficient DNA electrotransfer into tumors. AB - DNA transfer to tumor cells of antiproliferative genes or of genes coding for immunomodulatory or antiangiogenic products is a promising approach for cancer therapy. However, intratumoral injection of plasmid DNA either naked or associated to chemical vectors results in a low level of gene expression. Recently, electrically mediated gene transfer has been described to strongly increase foreign gene expression in various tissues. We confirm and extend these observations using long duration electric pulses for several murine and human tumor models, using a reporter gene encoding for luciferase. After plasmid intratumoral injection, eight electric pulses of 20-ms duration were delivered at a frequency of 1 Hz through two flat parallel stainless steel electrodes placed at each side of the tumor. Optimal gene transfer was obtained using a voltage-to distance ratio comprising between 400 and 600 V/cm. Two days after electrotransfer, we obtained a 10- to 1200-fold increase in gene expression over the naked DNA injection alone, leading to the expression of 0.6 to 300 ng luciferase per tumor. Moreover, histological results using beta-Gal reporter gene injected in H1299 tumor indicate that electrotransfer leads to a substantial increase in the percentage of beta-Gal positive cells. These results confirm the wide potential of electrotransfer for gene therapy in cancer. PMID- 11059582 TI - Testing of nonlinear electrofrictiophoresis in agarose gel. AB - It was theoretically predicted earlier that if a periodic force without constant component is applied to a particle, then the particle can produce a directed drift in some direction. The effect is named nonlinear electrofrictiophoresis, because it is crucial for its appearance that the friction force depends on the particle's velocity in a nonlinear manner. We test a possibility to observe this effect when a mixture of fragments of DNA (the DNA ladder) moves in the agarose gel. For this purpose, we study the nonlinear characteristics of a DNA ladder movement in the gel. The gels with the ladder were run under various electric field strengths. It was found that the friction coefficient for each DNA fragment in the ladder depends on the migration velocity, suggesting that energy dissipation during migration is a nonlinear function of velocity. This nonlinearity makes the system under consideration suitable for observing nonlinear electrofrictiophoresis. A possible velocity of directed drift under periodic electric drive without constant component was estimated numerically for experimentally observed dependencies. The velocity appeared to be comparable with that of migration under a constant field of moderate strength. A possible mechanism of energy dissipation during movement of DNA through the gel is discussed. PMID- 11059583 TI - Microstructuring of solid-supported lipid layers using SAM pattern generation by scanning electrochemical microscopy and the chemical lens. AB - To prepare patterns of adsorption sites for alkanethiols with high lateral resolution, we used the scanning electrochemical microscopy (SECM) to etch masks into uniform layers of nickel coated on gold surfaces. The patterning of the nickel mask was accomplished in aqueous solutions by electrogenerating nitric acid out of nitrite at an ultramicroelectrode. Due to the sluggish kinetics of nickel etching in acidic media, the pattern generated by a 10-microm tip was about 50-microm wide, depending on the duration of the etching. As an alternative, applying the principle of the chemical lens by adding potassium hydroxide as a scavenger, the size of the adsorption sites had been reduced to 4 microm, independent of the duration of etching. In a follow-up step, monolayers of 11-mercaptoundecanoic acid were formed on the exposed gold areas of the surface by self-assembly. Fluorescent liposomes containing tetramethylrhodamine labeled phospholipids were used to create solid-supported lipid layers (SSLLs). These fluorescent liposomes showed a selective binding affinity to the self assembled monolayers (SAMs) modified areas, but not to the nickel surface. The patterns generated were imaged by the SECM itself, as well as by optical and fluorescence microscopy. PMID- 11059584 TI - On-demand electrochemical release of DNA from gold surfaces. AB - Electrochemical quartz crystal microbalance (EQCM) is used to probe the electrochemically triggered release of nucleic acids from gold surfaces to solutions of physiological pH. The immobilization of nonthiolated DNA onto the gold surface is followed by an electrostatic desorption at -1.0 V (vs. Ag/AgCl). Steady-state frequency signals, corresponding to the removal of 261- and 644 ng/cm2 single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), respectively, were attained within 75 and 330 s. As expected for electrostatic repulsion, the amount released can be manipulated by tuning the potential. The small nonsteady state frequency signals observed at lower potentials indicate promise for a sustained DNA release. Applicability to gold ultramicroelectrodes (12.5-microm radius) is demonstrated in connection with voltammetric blocking experiments. We expect that such on-demand electrochemical release would be a useful addition to the arsenal of nonviral gene delivery routes. PMID- 11059585 TI - Revisiting spatial vision: toward a unifying model. AB - We report contrast detection, contrast increment, contrast masking, orientation discrimination, and spatial frequency discrimination thresholds for spatially localized stimuli at 4 degrees of eccentricity. Our stimulus geometry emphasizes interactions among overlapping visual filters and differs from that used in previous threshold measurements, which also admits interactions among distant filters. We quantitatively account for all measurements by simulating a small population of overlapping visual filters interacting through divisive inhibition. We depart from previous models of this kind in the parameters of divisive inhibition and in using a statistically efficient decision stage based on Fisher information. The success of this unified account suggests that, contrary to Bowne [Vision Res. 30, 449 (1990)], spatial vision thresholds reflect a single level of processing, perhaps as early as primary visual cortex. PMID- 11059587 TI - Upper-limit luminance for the surface-color mode appearance. AB - A series of experiments were carried out to reveal determinants for the mode of color appearance by measuring the upper-limit luminance of a color chip for the surface-color mode. We used a CRT color monitor to present test and surround stimuli in the surface-color mode. The stimuli were composed of a three-by-three array of color chips on a gray background with a white frame. The observer increased the luminance of a center test color until it just ceased to appear in the surface-color mode. Our results show that this upper-limit luminance was different among test colors, but their brightnesses, calculated from the luminance and brightness/ luminance values, were almost the same and were slightly below the brightness of the white frame. The existence of the surrounding color chips affected the results, but their sizes and spatial arrangements did not. When all of the luminances of the surrounds changed equally, the upper-limit luminances of the test colors for the surface-color mode appearance changed by the same ratio. This result indicates that the brightness of a target was a determinant for selecting the mode of color appearance and that the brightest surround stimulus acted as a cue for determining the judgment. PMID- 11059586 TI - Senescence of foveal and parafoveal cone sensitivities and their relations to macular pigment density. AB - Foveal and parafoveal increment thresholds were measured for 50 observers (12-88 years of age) under conditions that isolated retinal mechanisms dominated by short- (S-), middle- (M-), or long- (L-) wave-sensitive cones. Thresholds were obtained on the plateau of the threshold-versus-intensity function of each isolated mechanism and were referred to the retina by using individual measurements of ocular media and macular pigment density. Age-related increases in foveal thresholds, specified at the retina, were found for all three cone mechanisms. Parallel sensitivity losses for each cone mechanism were also observed at 4 degrees and 8 degrees in the temporal retina. A significant positive correlation was found between foveal macular pigment density and the S cone, but not the M- and L-cone, log sensitivity difference (0 degrees-8 degrees) specified at the retina. This relation is expected from the hypothesis that the macular pigment protects the photoreceptors from senescent losses in sensitivity. However, because this result is independent of age, it is interpreted as being due to local gain changes resulting from differential filtering of incident light by the macular pigment between the fovea and the parafovea. PMID- 11059588 TI - Probabilistic regularization in inverse optical imaging. AB - The problem of object restoration in the case of spatially incoherent illumination is considered. A regularized solution to the inverse problem is obtained through a probabilistic approach, and a numerical algorithm based on the statistical analysis of the noisy data is presented. Particular emphasis is placed on the question of the positivity constraint, which is incorporated into the probabilistically regularized solution by means of a quadratic programming technique. Numerical examples illustrating the main steps of the algorithm are also given. PMID- 11059589 TI - Shadow-invariant classification for scenes illuminated by daylight. AB - A physics-based method for shadow compensation in scenes illuminated by daylight is proposed. If the daylight is represented by a simplified form of the blackbody law and the camera filters are of infinitely narrow bandwidth, the relationship between red/blue (rm) and green/blue (gm) ratios as the blackbody's temperature changes is a simple power law where the exponent is independent of the surface reflectivity. When the CIE daylight model is used instead of the blackbody and finite bandwidths for the camera are assumed, it is shown that the power law still holds with a slight change to the exponent. This means that images can be transformed into a map of rm/gmA and then thresholded to yield a shadow independent classification. Exponent A can be precalculated from the CIE daylight model and the camera filter characteristics. Results are shown for four outdoor images that contain sunny and shadowed parts with vegetation and background. It is shown that the gray-level distributions of the pixels in the transformed images are quite similar for a given component whether or not it is in shadow. The transformation leads to bimodal histograms from which thresholds can easily be selected to give good classifications. PMID- 11059590 TI - Noise-residue filtering of interferometric phase images. AB - The effect of noise-induced phase inconsistency on interferometric phase information content is studied. Phase inconsistencies, or residues, hinder a correct unwrapping of the phase signal. The probability of noise-induced phase inconsistencies is obtained as a function of the signal-to-noise ratio. Moreover, a two-dimensional noise-residue filter, intended to be applied as a preprocessing step before phase unwrapping, is proposed. The method is based on the observation that the noise creates adjacent phase inconsistencies mainly in interferometric phase images. A local analysis leads to a set of rules to be applied to reduce noise-induced phase inconsistencies. The filter performances are tested on noisy synthetic and real phase data. PMID- 11059591 TI - Spectral model of a fluorescent ink halftone. AB - A model is presented of a fluorescent ink halftone. Unlike a nonfluorescent ink, which only absorbs, a fluorescent ink absorbs higher-energy photons and emits lower-energy photons. The amount of fluorescent light produced depends on the percent absorption of the incident light. For fluorescent ink printed on paper, both photon scattering within the paper substrate and multiple internal reflections between the ink layer and the paper substrate significantly increase the percent absorption, so a realistic model must include these effects. The model presented here utilizes the generalized Clapper-Yule theory, which accounts for photon diffusion that is due to both scatter and internal reflection. It is shown that while multiple internal reflections alone only marginally increase the percent absorption, when there are both scattering and internal reflection, the percent absorption is increased significantly. The current study is a theoretical model and does not present experimental results. PMID- 11059592 TI - Optimal processors for images with an arbitrary number of gray levels. AB - We present a new group of processors, optimal in a maximum-likelihood sense, for target location in images with a discrete number of gray levels. The discrete gray-level distribution can be of any arbitrary form. We compare the performance of the processor derived for five discrete levels with the performance of a processor derived for a continuous Gaussian distribution and show that there are cases when only the processor derived for discrete levels will exhibit satisfactory performance. We give an explanation of this difference based on moment analysis and show how the correlation orders are related to statistical moments of the input scene. PMID- 11059593 TI - Cone-beam reconstruction by backprojection and filtering. AB - A new analytical method for tomographic image reconstruction from cone-beam projections acquired on the source orbits lying on a cylinder is presented. By application of a weighted cone-beam backprojection, the reconstruction problem is reduced to an image-restoration problem characterized by a shift-variant point spread function that is given analytically. Assuming that the source is relatively far from the imaged object, a formula for an approximate shift invariant inverse filter is derived; the filter is presented in the Fourier domain. Results of numerical experiments with circular and helical orbits are considered. PMID- 11059594 TI - General propagation equation of flattened Gaussian beams. AB - The closed-form propagation equation of flattened Gaussian beams passing through a paraxial optical ABCD system, in which the linear gain and absorption media are included, is derived, and its general applicable advantage is illustrated with numerical examples. PMID- 11059595 TI - Beam propagation properties of radial laser arrays. AB - A detailed study of phase-locked and non-phase-locked radial laser arrays is presented. The closed-form propagation expressions for the beamlets and the resulting beam are given, which enable us to study beam propagation properties of radial laser arrays for both phase-locked and non-phase-locked cases. Numerical calculation examples are given to illustrate the application of our analytical results and the differences between phase-locked and non-phase-locked radial arrays. PMID- 11059596 TI - Propagation of Laguerre-Bessel-Gaussian beams. AB - New exact solutions to the paraxial wave equation are obtained in the form of a product of Laguerre polynomials, Bessel functions, and Gaussian functions. In the limit of large Laguerre-Gaussian beam size, the Bessel factor dominates and the solution sets reduce to the modes of closed resonators, hollow metal waveguides, and dielectric waveguides. In the opposite limit the solutions reduce to Laguerre Gaussian modes of open resonators and graded-index waveguides. These solutions are valid for electromagnetic waves traveling through free space, and they are valid for propagation through circularly symmetric optical systems representable by ABCD matrices as well. An interesting feature of the new solution set is the existence of three mode indices, where only two are required for an orthogonal expansion. As an example, Laguerre-Gaussian beam propagation through an optical system that contains a Bessel-like amplitude filter is discussed. PMID- 11059597 TI - Propagation-induced polarization changes in partially coherent optical beams. AB - Propagation of a partially coherent optical beam inside a linear, nondispersive, dielectric medium is studied, taking into account the vector nature of the electromagnetic field. Propagation-induced polarization changes are studied by using the Gaussian-Schell model for the cross-spectral-density tensor. The degree of polarization changes with propagation and also becomes nonuniform across the beam cross section. The extent of these changes depends on the coherence radius associated with the cross-correlation function. For optical beams with symmetric spectra, the bandwidth of the source spectra is found to play a relatively minor role. PMID- 11059598 TI - Low-coherence interferometry in random media. I. Theory. AB - We present a new nonperturbative theoretical method for the analytical description of light propagation in random multiscattering media. The method is illustrated through the calculation of an expression that describes optical backscattering from a semi-infinite disordered medium. A companion paper [J. Opt. Soc. Am. A 17, 2034 (2000)] compares the theoretical expression with experimental data. PMID- 11059599 TI - Low-coherence interferometry in random media. II. Experiment. AB - We present experimental results of measurements of light backscattering from semi infinite disordered media by low-coherence interferometry (LCI). These results are compared with the theory developed in part I [J. Opt. Soc. Am. A 17, 2024 (2000)]. A comparison of the experimental data with the theoretical formulas based on the coherent phase approximation allows us to extract substantial information about the structure of the studied media. Our results demonstrate that LCI is an effective optical technique for studying nonuniform media even in the case in which the dimensions of nonuniformities are much less than the wavelength of the scattered light. PMID- 11059600 TI - Features in coherent transmittance of a monolayer of particles. AB - Coherent and incoherent transmittance values of a monolayer of particles are considered. Such a monolayer is a set of particles whose centers are located in the same plane. We set forth the conditions for the effect of coherent transmittance quenching, which takes place as a result of the interference between incident and forward-scattered waves. Using the single-scattering approximation we determined size parameters and particle refractive indexes for this interference effect in the case of identical isotropic spherical particles. The influence of polydispersity and the fine structure of light-scattering characteristics on the quenching effect has been estimated. It is shown that the polydispersity destroys this interference effect only at large widths of particle size distribution functions. The influence of multiple scattering on this effect is considered in the quasi-crystalline approximation. Multiple scattering results in increasing size parameters and decreasing particle concentration at which coherent transmittance quenching takes place in comparison with the case of single scattering. Our theoretical results for suspensions of latex particles in water are in fairly good agreement with the experimental results. PMID- 11059601 TI - Real-space Green's function calculation for the solution of the diffusion equation in stratified turbid media. AB - We have derived the space-time Green's function for the diffusion equation in layered turbid media, starting from the case of a planar interface between two random scattering media. This new approach for working directly in real space permits highly efficient numerical processing, which is a decisive criterion for the feasibility of the inverse problem in biomedical optics. The results obtained by this method in the case of a two-layered medium are compared with Monte Carlo simulations. PMID- 11059602 TI - Cross talk in holographic memories with lensless phase-conjugate holograms. AB - Recently holographic memory with lensless phase-conjugate holograms has attracted much attention because it opens up the possibility of compact holographic memories. We investigate cross-talk noise in compact holographic memories with angular multiplexing. It turns out that the optimum angular separation is the same as that for the Fourier plane hologram in the leading order and that the noise-to-signal ratio is independent of the positions in the output plane, similar to the case of the image plane hologram. PMID- 11059603 TI - Phase-shift calibration algorithm for phase-shifting interferometry. AB - We propose a novel phase-shift calibration algorithm. With this technique we determine the unknown phase shift between two interferograms by examining the sums and differences of the intensities on each interferogram at the same spatial location, i.e., I1(x, y) +/- I2(x, y). These intensities are normalized so that they become sinusoidal in form. A uniformly illuminated region of the interferograms that contains at least a 2pi variation in phase is examined. The extrema of these sums and differences are found in this region and are used to find the unknown phase shift. An error analysis of the algorithm is provided. In addition, an error-correction algorithm is implemented. The method is tested by numerical simulation and implemented experimentally. The numerical tests, including digitization error, indicate that the phase step has a root-mean-square (RMS) phase error of less than 10(-6) deg. Even in the presence of added intensity noise (5% amplitude) the RMS error does not exceed 1 deg. The accuracy of the technique is not sensitive to nonlinearity in the interferogram. PMID- 11059604 TI - Depolarization and retardation of a birefringent slab. AB - Mueller matrices for normal transmission of light through a birefringent slab are formulated to analyze retardation and depolarization. A finite wave band, wedge slab, and microroughness may cause a spread in retardance, which in turn produces depolarization. The spectra of depolarization, cross-polarized transmittance, and retardance by rotating-analyzer ellipsometry are simulated for the quasi monochromatic effect with a finite bandwidth. These spectra agree excellently with the measured spectra for a sapphire slab. The depolarization spectrum simulated for the wedge effect fits the measured spectrum in the long-wave region but is too small in the short-wave region. The depolarization simulated for incoherent multiple reflections demonstrates the oscillating structure, which is small compared with the measured depolarization. The finite bandwidth effect contributes more than the other effects to the measured depolarization of a sapphire slab. PMID- 11059605 TI - Generalized formulation and symmetry properties of reciprocal nonabsorbing polarization devices: application to liquid-crystal displays. AB - We present a general formulation based on the Jones-matrix theory for reciprocal nonabsorbing polarization devices, including polarization interference filters and liquid-crystal displays. The development of this formulation is based on general symmetry conditions that relate the Jones matrix when the device is illuminated from the front side and from the back side. The application to liquid crystal displays results in a constraint of the Jones-matrix elements, which represents a generalization of the existing models that explain their modulation properties. PMID- 11059606 TI - Focusing of electromagnetic waves by paraboloid mirrors. I. Theory. AB - We derive a solution to the problem of a plane electromagnetic wave focused by a parabolic mirror. The solution is obtained from the Stratton-Chu integral by solving a boundary-value problem. Our solution can be considered self-consistent. We also derive the far-field, i.e., Debye, approximation of our formulas. The solution shows that when the paraboloid is infinite, its focusing properties exhibit a dispersive behavior; that is, the structure of the field distribution in the vicinity of the focus strongly depends on the wavelength of the illumination. We show that for an infinite paraboloid the confinement of the focused energy worsens, with the energy distribution spreading in the focal plane. 2000 Optical Society of America [S0740-3232(00)01309-0] OCIS codes: 260.0260, 260.2110, 050.1960, 260.5430. PMID- 11059607 TI - Focusing of electromagnetic waves by paraboloid mirrors. II. Numerical results. AB - We present results of numerical computations obtained from a theory described in Part 1 of our current investigations [J. Opt. Soc. Am. A 17, 2081 (2000)]. We show that a segment of a paraboloid mirror produces an intensity distribution identical to that of a high-aperture lens. It is shown that when the convergence angle of the paraboloid is increased beyond the pi/2 limit, the lateral resolution in the direction orthogonal to the incident polarization improves, whereas in the other direction the resolution worsens. Numerical results show that paraboloid mirrors of high convergence angle exhibit dispersion; that is, when the focal length is altered by a quarter of the wavelength the intensity in the focus changes from its maximum to its minimum value. A focal shift is observed that, in the case of a paraboloid of low convergence angle is identical to the Fresnel shift. However, a focal shift is also observed at large convergence angles. PMID- 11059608 TI - Cavity phase engineering for stable enhanced terahertz pulse trains. AB - We show that the cavity round-trip Gouy phase leads to pulse-to-pulse variation of the absolute phase and temporal profile of circulating few- or single-cycle pulses in empty resonators. This pulse-to-pulse variation can be eliminated by the proper insertion of a lens into the cavity. An application to terahertz resonators with phase-locked feedback is discussed. PMID- 11059609 TI - Derivation of a detectability index for correlated responses in multiple alternative forced-choice experiments. AB - In a recent paper [J. Opt. Soc. Am. A 17, 206 (2000)], a modified detectability index, d'r, was proposed to accommodate correlations between internal responses in multiple-alternative forced-choice (MAFC) experiments. The derivation given in that work pertained only to two-alternative forced choice, although it was shown empirically that the result held for general MAFC tasks when the correlation between responses is constant. Here we present a rigorous derivation that shows that the d'r result generalizes to MAFC tasks in this case. PMID- 11059610 TI - Poincare sphere representation of the fixed-polarizer rotating-retarder optical system. AB - The trajectory of the polarization state of a monochromatic light beam after it passes through a fixed linear polarizer and a rotating linear retarder of arbitrary retardance delta is determined on the Poincare sphere. The three dimensional figure-8 contour is shown to be the line of intersection of a right circular cylinder with the sphere. The cylinder is parallel to the polar (S3) axis, touches the sphere at the equator (at the point that represents the linear polarization transmitted by the fixed polarizer), and has a radius r = sin2(delta/2). Projections of the trajectory in the coordinate planes of the normalized Stokes parameter space (s1, s2, S3) are also determined. PMID- 11059611 TI - Diagonal versus affine transformations for color correction. AB - Standard methods for color correction involve the use of a diagonal-matrix transformation. Zaidi proposes the use of a two-parameter affine model; we show that this offers no improvement in terms of accuracy over the diagonal model, especially if a sharpening transformation is also used. PMID- 11059612 TI - Influence of right atrial structure on outcome of radio-frequency catheter ablation for common atrial flutter. AB - Radiofrequency catheter ablation (RFCA) targeting the cavotricuspid isthmus is usually an effective treatment for common atrial flutter (AFL), except in a small subset of patients and the reason for this has yet to be elucidated. The present study investigated the relationship between the outcome of RFCA for common AFL and the anatomy of the right atrium as seen on angiography. Twenty consecutive patients who underwent RFCA for common AFL were divided into 2 groups according to the results of RFCA. Group A comprised 13 patients whose AFL was abolished, fulfilling the criteria of success by the conventional catheter approach, and group B comprised 7 patients whose AFL could not be abolished according to the criteria for success (n=4) or was abolished following an additional superior vena cava approach (n=3). On angiography, the cavotricuspid isthmus was longer (3.5+/ 0.5 vs 2.2+/-0.6 cm) and deeper (0.94+/-0.35 vs 0.49+/-0.19 cm) in group B than in group A (both p<0.01). The height of the eustachian valve was also greater in group B than in group A (1.4+/-1.1 vs 0.48+/-0.48 cm, p<0.02). These results suggest that the anatomical structure of the cavotricuspid isthmus affects the outcome of RFCA for common AFL. PMID- 11059613 TI - Assessment of aortic atherosclerosis and carotid atherosclerosis in coronary artery disease. AB - The present study investigated the relationship between aortic atherosclerosis and carotid atherosclerosis, and studied the effects of coronary risk factors for these arteries. The subjects consisted of 78 patients with coronary artery disease (CAD) and 69 patients without CAD. All subjects underwent enhanced computed tomography and B-mode ultrasonography within a short time period to determine the extent of aorta and carotid atherosclerosis. Significant correlations between maximal aortic wall thickness (MAWT) and aortic wall volume (AWV) with carotid intima-media thickness (IMT) were demonstrated. MAWT, AWV and IMT were significantly higher in patients with CAD compared with controls (p=0.009, p=0.024, p=0.001, respectively). Furthermore, there were significant differences in MAWT, AWV and IMT among groups classified by the number of coronary artery stenoses, and no significant differences among groups classified by risk factors, but it was shown that MAWT, AWV and IMT increased gradually as the risk factors increased in number. MAWT, AWV and IMT had positive correlations with age, systolic blood pressure and triglyceride, and a negative correlation with high density lipoprotein-cholesterol. This study demonstrated that both aortic atherosclerosis and carotid atherosclerosis are closely correlated with coronary atherosclerosis, and that the atherosclerosis indices are independently associated with age and hyperlipidemia. PMID- 11059614 TI - Effect of a single oral dose of pilsicainide on pacing thresholds in pacemaker patients with and without paroxysmal atrial fibrillation. AB - A single oral dose of pilsicainide, a class 1c antiarrhythmic drug, is effective in terminating acute-onset atrial fibrillation (AF), but its effect on pacing thresholds in pacemaker patients is unknown. The present study measured atrial and ventricular pacing thresholds after a single oral dose of pilsicainide in patients with and without AF. Twelve patients with dual-chamber pacemakers were evaluated. Pacing thresholds as well as plasma pilsicainide concentration were measured prior to and then at 30, 60, 90, 120 and 180 min and 24h following a single oral dose of pilsicainide (150 mg). Six patients had paroxysmal AF and the remaining 6 did not. Pacing thresholds increased significantly (134+/-8%) in the atrium (p<0.05) and in the ventricle (155+/-11%; p<0.001) following pilsicainide administration in all 12 patients. Plasma concentrations of pilsicainide showed a positive liner correlation with pacing thresholds (R=0.62, p<0.0001 in the atrium; R=0.74, p<0.0001 in the ventricle). Atrial pacing thresholds in the patients with AF showed a significant increase at 90, 120 and 180 min compared with the patients without AF (p<0.05). There was no significant difference in either the ventricular pacing threshold or the plasma pilsicainide concentration in the patients with and without AF. It was concluded that a single oral dose of pilsicainide increases the pacing thresholds in both the atrium and ventricle in a selected group of pacemaker-implanted patients; that is, those who are aged and with AF. Thus, careful attention should be paid to pacemaker-dependent patients, particularly those with paroxysmal AF, when administering pilsicainide. PMID- 11059615 TI - Levels of serum interleukin-10 reflect disease activity in patients with cardiac sarcoidosis. AB - Cardiac involvement is the major determinant of morbidity and mortality in patients with sarcoidosis, but clinical evaluation of the disease activity is occasionally difficult in cardiac sarcoidosis. The present study examined whether serum levels of interleukin-10 (IL-10) could reflect the disease activity of patients with cardiac sarcoidosis. Serum IL-10 levels were measured using an enzyme-linked immunosorbent assay, and compared with clinical manifestation, levels of angiotensin-converting enzyme (ACE), levels of lysozyme and accumulation of gallium-67 citrate. Sera were collected from 8 patients with cardiac sarcoidosis (CS group), 22 patients with miscellaneous heart diseases except for sarcoidosis (MHD group), and 8 healthy control subjects (HC group). Serum IL-10 levels of the CS group were significantly higher than those of the 2 control groups. Before steroid therapy, the levels of IL-10 in the CS group showed a significantly positive correlation with levels of ACE (r=0.868, p<0.05) and lysozyme (r=0.890, p<0.05). In 5 patients who were analyzed before and after steroid therapy, the levels of IL-10 tended to correlate with a decrease of an abnormal accumulation in gallium-67 citrate. Serum IL-10 levels may play a role in evaluation of the disease activity in patients with cardiac sarcoidosis. PMID- 11059616 TI - Evaluation of flow volume and flow patterns in the patent false lumen of chronic aortic dissections using velocity-encoded cine magnetic resonance imaging. AB - In 21 patients with chronic aortic dissections and proven patent false lumens, the flow volume and flow patterns in the patent false lumens was evaluated using velocity-encoded cine magnetic resonance imaging (VENC-MRI) and the relationship between the flow characteristics and aortic enlargement was retrospectively examined. Flow patterns in the false lumen were divided into 3 groups: pattern A with primarily antegrade flow (n=6), pattern R with primarily retrograde flow (n=3), and pattern B with bidirectional flow (n=12). In group A, the rate of flow volume in the false lumen compared to the total flow volume in true and false lumens (%TFV) and the average rate of enlargement of the maximum diameter of the dissected aorta per year (deltaD) were significantly greater than in groups R and B (%TFV: 74.1+/-0.07 vs 15.2+/-0.03 vs 11.8+/-0.04, p<0.01; deltaD: 3.62+/-0.82 vs 0 vs 0.58+/-0.15 mm/year, p<0.05, respectively). There was a significant correlation between %TFV and deltaD (r=0.79, p<0.0001). Evaluation of flow volume and flow patterns in the patent false lumen using VENC-MRI may be useful for predicting enlargement of the dissected aorta. PMID- 11059617 TI - Plasma cardiac natriuretic peptides as biochemical markers of recurrence of atrial fibrillation in patients with mild congestive heart failure. AB - To determine changes in plasma brain natriuretic peptide (BNP) after direct current cardioversion (DC) and to evaluate the relationship between plasma atrial natriuretic peptide (ANP) and BNP and the recurrence of atrial fibrillation (AF) after DC in patients with mild congestive heart failure (CHF), plasma ANP and BNP were measured before and after DC in 71 patients with mild CHF and then followed. In 65 patients with successful DC, both ANP and BNP decreased 15 min after DC. Cox stepwise multivariate analysis among 14 variables such as age, history of AF, echocardiographic parameters, medication and ANP and BNP revealed that only low ANP (p=0.005) and high BNP before DC (p=0.0002) were independent predictors of recurrent AF. A ratio of ANP to BNP less than 0.44 was a significant risk factor for AF recurrence by Kaplan-Meier analysis (p=0.02). BNP began to decrease immediately after successful DC. High BNP and relatively low ANP compared with BNP were independent risk factors of AF recurrence in patients with mild CHF. PMID- 11059618 TI - Effect of a novel cardioprotective agent, JTV-519, on metabolism, contraction and relaxation in the ischemia-reperfused rabbit heart. AB - The effect of a novel cardioprotective agent, JTV-519 on myocardial metabolism and contraction during ischemia and reperfusion was investigated by means of phosphorus 31-nuclear magnetic resonance (31P-NMR) in Langendorff rabbit hearts. Normothermic, 20-min, global ischemia was followed by 30-min of postischemic reperfusion and JTV-519 was administered from 40 min prior to the global ischemia. Adenosine triphosphate (ATP), creatine phosphate (PCr), inorganic phosphate (Pi), intracellular pH (pHi), left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP) and coronary flow were measured. Fourteen hearts were divided into 2 experimental groups of 7: Group I were controls and Group II were perfused with JTV-519 (10(-6) mol/L). During ischemia, Group II showed a significant (p<0.01) inhibition of the increase in Pi and LVEDP and the decrease in ATP and pHi, compared with Group I. After postischemic reperfusion, Group II also showed a significant (p<0.01) improvement in ATP and pHi as compared with Group I. There were no differences in LVDP or coronary flow during ischemia and reperfusion between the 2 groups. In conclusion, JTV-519 had a significant beneficial effect on myocardial energy metabolism and relaxation during both myocardial ischemia and reperfusion. PMID- 11059619 TI - Comparison of the electropharmacological effects of verapamil and propranolol in the halothane-anesthetized in vivo canine model under monophasic action potential monitoring. AB - The cardiovascular profile of verapamil was assessed in the halothane anesthetized canine model and compared with that of propranolol. Verapamil was infused at the rates of 1, 3 and 10 microg x kg(-1) x min(-1) (n=6), whereas propranolol was administered at a fixed rate of 10 microg x kg(-1) x min(-1) (n=6). Each infusion was performed over 30 min, and the parameters were assessed for 20-30 min after the start of each infusion. Verapamil in a dose of 10 microg x kg(-1) x min(-1) significantly suppressed atrio-ventricular (AV) node conduction and slightly decreased the mean blood pressure, but no significant change was detected in the left ventricular end-diastolic pressure, maximum upstroke velocity of the left ventricular pressure, sinus automaticity, double product, cardiac output, intraventricular conduction, and ventricular repolarization phase and refractoriness. Propranolol suppressed AV node conduction to an extent similar to that of verapamil, but it also inhibited intraventricular conduction, sinus automaticity and ventricular contraction, increased the ventricular refractoriness, and decreased the double product and cardiac output, without any significant change in the other variables measured. These results suggest that verapamil can selectively affect the AV node, and that the greater part of the suppressive action of propranolol on the multiple cardiovascular performance is through a beta-blocking action and direct membrane effect, although the halothane inhalation itself might have modified each of the drug's effects. The abbreviation of the relative refractory period of the ventricle by propranolol may show its potential utility for re-entry type ventricular tachycardia. PMID- 11059620 TI - Primary leiomyosarcoma of the pulmonary artery mimicking massive pulmonary thromboembolism. AB - A 57-year-old man with dyspnea and dry cough exhibited pulmonary embolism. Pulmonary arteriography demonstrated absent perfusion of the left main and the right upper and middle lobe pulmonary arteries. A diagnosis of chronic pulmonary thromboembolism was assumed and surgical thromboendarterectomy was attempted under standard cardiopulmonary bypass. At operation, a tumor had invaded far into both the right and left pulmonary arteries and radical resection was impossible. The final pathological diagnosis was primary leiomyosarcoma of the pulmonary artery. The patient refused any adjuvant therapy and died 63 days after the surgery. The clinical presentation of this case was similar to that of pulmonary thromboembolism and its diagnosis and treatment were very difficult. PMID- 11059621 TI - Dexamethasone-induced cardiogenic shock rescued by percutaneous cardiopulmonary support (PCPS) in a patient with pheochromocytoma. AB - A 52-year-old man with pheochromocytoma had cardiogenic shock and was rescued using a percutaneous cardio pulmonary supporting system. After recovery, diagnostic tests including metaiodobenzylguanidine scintigraphy and computed tomography, revealed the pheochromocytoma which was confirmed by histology. It was postulated that the acute episode was induced by intra-joint dexamethasone, which increased the production of epinephrine and augmented the sensitivity of cardiomyocytes for catecholamine, thereby inducing the cardiomyopathy. PMID- 11059622 TI - Coronary microvascular abnormality in the reversible systolic dysfunction observed after noncardiac disease. AB - Acute reversible left ventricular wall motion abnormalities mimicking myocardial stunning have been reported with noncardiac disease and their coronary angiograms did not demonstrate organic stenosis or vasospasm in the epicardial coronary arteries. Thus, this mechanism has not yet been fully clarified. Two patients are reported as demonstrating acute reversible wall motion abnormalities after noncardiac disease. The electrocardiographic and echocardiographic findings mimicked myocardial stunning and confirmed the previous reports. The coronary angiograms did not show any corresponding coronary stenosis or vasospasm, but did show a reduced coronary flow reserve. Cardiac metaiodobenzylguanidine scintigraphy demonstrated regional defects involving the apex, a decreased heart/mediastinum ratio and an enhanced washout rate, which partially returned to normal after 3 months. Microvascular dysfunction and sympathetic nervous abnormalities might be responsible for the reversible contractile impairment. PMID- 11059623 TI - Atrial double potentials associated with the elimination of the electrical connection between the coronary sinus (CS) and the left atrium in two cases of Wolff-Parkinson-White syndrome with a CS-connected accessory pathway. AB - A curious retrograde conduction in connection with the coronary sinus (CS) musculature was observed in 2 patients. After the failed ablation procedure, the atrial electrogram during ventricular pacing presented double potentials, the first component of which was sharp and with an activation sequence that was the same before ablation (CS distal to proximal). The second component of the double potentials was dull and had a decremental property; its activation sequence was in reverse (proximal to distal). In both cases, the first component disappeared after successful ablation. These findings suggest that the first component was the CS electrogram conducted over the accessory pathway and the second component was the left atrial electrogram conducted through the inter-atrial septum. The separation of each electrogram is probably the result of a block between the accessory pathway connected to the CS musculature and the left atrium. These are unusual cases of an accessory pathway connected to the CS musculature, which separates the left atrial myocardium at the distal portion from the ostium. PMID- 11059624 TI - Papillary fibroelastoma in association with thrombosis on a mechanical valve. AB - A patient with a thrombosed mechanical valve underwent valve re-replacement during which a tumor of the left ventricular outflow tract with the typical macroscopic and microscopic characteristics of a papillary fibroelastoma was successfully removed surgically. The 60-year-old woman had undergone isolated mitral valve replacement with a St Jude Medical 29-mm valve for mitral regurgitation 15 years ago. The present admission was for investigation of dyspnea on exertion. Two-dimensional transthoracic echocardiography demonstrated a posteroseptal, pedunculated mass, measuring 1.3x1.0 cm, in the outflow tract of the left ventricle, mild mitral regurgitation and slight aortic stenosis. PMID- 11059625 TI - Intracoronary positive and negative contrast jets at a 'napkin ring' coronary stenosis: a case report. AB - A very short coronary stenosis is somewhat difficult to correctly evaluate by conventional coronary angiography. The demonstration of positive and negative contrast jets observed just distal to the lesion is helpful in identifying the severity of the disease causing this disorder. In the present case, intravascular ultrasound confirmed the presence of a very short stenosis and membranous disease at the site of the lesion. The presence of intracoronary contrast jets reflects the morphology and severity of disease of the membranous type, although the reason for the appearance of contrast jets with this type of lesion should be sought. PMID- 11059626 TI - Coronary stent implantation in a septal perforator artery: case report and review of the literature. AB - Septal perforator arteries play an important role in the blood supply of the anterior interventricular septum. Their intramyocardial course makes them inaccessible for coronary bypass revascularization. Although modern catheter based techniques might be superior to coronary bypass grafting in offering the most complete revascularization in selected patient populations, a systematic review of the literature revealed a paucity of data regarding the outcome of these patients. The present report describes coronary stent implantation in a dominant septal perforator artery and the analysis of the anatomic relationship between the stent and the intraventricular septum using a new imaging technique, catheter-based intracardiac ultrasound. PMID- 11059627 TI - Circulating hepatocyte growth factor as a marker of thrombus formation in unstable angina pectoris. AB - A new enzyme-linked immunosorbent assay can detect 10 pg/ml of human hepatocyte growth factor (HGF). Circulating HGF was significantly higher in patients with unstable angina (296+/-184 pg/ml, mean+/-SD, n=36) than in healthy volunteers (201+/-64 pg/ml, n=250, p<0.0001). Individual concentrations exceeded the mean control value +2 SD (329 pg/ml) in 12 of the 36 (33%) patients with unstable angina. The present study indicates that this new, sensitive HGF assay can successfully detect thrombosis in patients with unstable angina. PMID- 11059628 TI - The renin-angiotensin system and cardiovascular disease. AB - Suppression of the renin-angiotensin system (RAS) with angiotensin-converting enzyme (ACE) inhibitors is an established method for controlling blood pressure and reducing the risk of cardiovascular disease. In addition to reducing blood pressure, suppression of the RAS is able to protect against the target-organ damage that results from hypertension. Unfortunately, despite the use of ACE inhibitors and agents from the other classes of conventional antihypertensives, effective control of blood pressure remains poor. A major contribution to this failure to control blood pressure appears to be lack of compliance with the prescribed medication, arising from the presence of unacceptable side effects. Angiotensin II type 1 (AT1) receptor blockers, such as candesartan, are the latest class of antihypertensive agent to be developed. They target the AT1 receptor - the final common pathway for all the known negative cardiovascular effects of angiotensin II - and provide pronounced antihypertensive efficacy without the side effects of cough and angioneurotic oedema that are associated with the use of ACE inhibitors. PMID- 11059629 TI - Angiotensin II type 1 receptor blockade: a novel therapeutic concept. AB - Angiotensin II type 1 (AT1) receptor blockers, such as candesartan, are attractive alternatives to ACE inhibitors in the treatment of hypertension and cardiovascular disease. Although angiotensin-converting enzyme (ACE) inhibitors are able to suppress the renin-angiotensin system (RAS), their mechanism of action may limit their clinical utility in the treatment of hypertension. For example, they act as competitive inhibitors of ACE. This means that their effects can be overcome by high levels of angiotensin I, which occur after ACE inhibition due to removal of the negative feedback effect of angiotensin II on renal renin release. ACE inhibitors are also unable to block the production of angiotensin II by non-ACE-mediated pathways. Furthermore, ACE is not a specific enzyme. Its inhibition therefore has effects on other substances, such as bradykinin, leading to the class-specific side effects associated with ACE inhibitors. Candesartan, on the other hand, binds insurmountably to the AT1-receptor, thereby providing more complete blockade of the negative cardiovascular effects of angiotensin II than is possible with ACE inhibitors. The specificity of AT1-receptor blockade also ensures that efficacy is achieved without inducing the side effect of cough that results from the non-specific consequences of ACE inhibition. Preclinical and early clinical studies demonstrate that AT1-receptor blockers produce at least the same degree of target-organ protection as has been demonstrated for ACE inhibitors. Additional benefits of AT1-receptor blockers may arise from the fact that, unlike ACE inhibitors, they do not prevent the activity of angiotensin II on AT2-receptors in the heart, which is thought to reduce cardiac remodelling. From a mechanistic perspective, therefore, AT1-receptor blockers appear to have advantages over ACE inhibitors, in terms of a more complete blockade of angiotensin II effects, while also avoiding the specific side effects associated with ACE inhibition. PMID- 11059630 TI - Differences among angiotensin II type 1 receptor blockers: characteristics of candesartan cilexetil. AB - Several angiotensin II type 1 (AT1) receptor blockers are now available for the treatment of hypertension. Although the agents in this class all act by blocking the AT1-receptor, they differ in their pharmacokinetics and binding characteristics. One of the newest AT1-receptor blockers, candesartan cilexetil, is administered in an inactive form and is rapidly and completely converted to the active drug, candesartan, during gastrointestinal absorption. In vitro studies have shown that candesartan has the highest receptor affinity of all the available AT1-receptor blockers and is not displaced from the receptor by high concentrations of angiotensin II. The tight and long-lasting binding of candesartan to the AT1-receptor provides effective blockade of the negative cardiovascular effects of angiotensin II. PMID- 11059631 TI - Improving antihypertensive efficacy while maintaining placebo-like tolerability. AB - Potency and efficacy, duration of action, organ-specific effects and tolerability are the main considerations when choosing among antihypertensive therapies. Candesartan has been shown in in vitro animal models to bind insurmountably to the angiotensin II type 1 (AT1) receptor, thus providing effective blockade of all the major negative cardiovascular effects of angiotensin II. Its binding characteristics differentiate candesartan from other AT1-receptor blockers. Candesartan cilexetil has been found to produce a predictable and pronounced dose dependent decrease in blood pressure, with placebo-like tolerability even at the highest doses studied. In comparison with the standard 50-mg dose of losartan, candesartan cilexetil, 16 mg, was significantly more effective in suppressing the renin-angiotensin system and in reducing trough diastolic blood pressure. Pooled results from placebo-controlled trials also indicate that candesartan cilexetil has equivalent efficacy to irbesartan. In addition, the extent of blood pressure lowering by candesartan cilexetil has been shown to be similar to that of agents in the other major classes of antihypertensive drugs, and to be effective in combination therapy with diuretics and calcium channel blockers. Candesartan cilexetil combines 24-h blood pressure lowering with placebo-like tolerability and is therefore an important advance in antihypertensive therapy. PMID- 11059632 TI - Achieving quality 24-h blood pressure control with candesartan cilexetil. AB - Epidemiological evidence suggests that optimal blood pressure control requires strategies that lower blood pressure consistently and fully throughout 24 h. In order to maximize compliance, antihypertensive agents also need to be well tolerated and effective when administered at a convenient once-daily dose. The new angiotensin II type 1 (AT1) receptor blocker candesartan binds tightly to, and dissociates slowly from, the AT1-receptor and thereby provides long-lasting suppression of the renin-angiotensin system. This is likely to explain its pronounced antihypertensive efficacy, which is maintained smoothly over 24 h. The trough-to-peak ratio is a useful measure of the persistence of antihypertensive efficacy at the end of the dosing interval. This ratio was found to be close to the ideal of 1.0 for candesartan cilexetil, 8 and 16 mg, whereas it was 0.7 for the prototype AT1-receptor blocker losartan, 50 mg. The antihypertensive effect of candesartan cilexetil, 16 mg, was also significantly greater than that of losartan, 100 mg, as demonstrated by ambulatory blood pressure measurements 0-36 h after dosing and by clinic measurements 48 h after dosing. By controlling blood pressure well beyond the normal dosing interval, candesartan cilexetil provides cardiovascular protection even in those patients who may occasionally miss doses. PMID- 11059633 TI - Efficacy and tolerability of candesartan cilexetil in special patient groups. AB - Patients with hypertension do not comprise a homogeneous group, and the majority present with a variety of concomitant and associated conditions. Antihypertensive therapies should therefore be effective and well tolerated in a wide range of patients and should, ideally, ameliorate the negative target-organ effects of hypertension, such as atherosclerosis, cardiovascular remodelling and renal impairment. Evidence is accumulating that the new angiotensin II type 1 receptor blocker, candesartan cilexetil, lowers blood pressure effectively and is well tolerated in a variety of patient groups, including women and the elderly. In patients with severe hypertension, a treatment schedule based on candesartan cilexetil, with the addition of diuretic and calcium antagonist therapy as needed, has been found to control blood pressure successfully. Candesartan cilexetil does not affect glucose tolerance or lipid profiles in patients with diabetes mellitus, and it is not associated with any of the side effects of other antihypertensive agents that would make it unsuitable for use in patients with pulmonary disease. Initial clinical studies have indicated that candesartan cilexetil is well tolerated and effective in patients with heart failure. Furthermore, the available evidence shows that treatment with candesartan cilexetil can reverse the negative effects of hypertension on left ventricular hypertrophy and microalbuminuria. It therefore appears that the pronounced efficacy and placebo-like tolerability of candesartan cilexetil, as demonstrated in large clinical trials of patients with mild to moderate hypertension, can be extended to a wide range of specific patient groups. PMID- 11059634 TI - Improving prognosis in hypertension: exploring the benefits of angiotensin II type 1 receptor blockade. AB - Prognosis can be improved in hypertensive patients not only by reducing blood pressure, but probably also by effective suppression of adverse neurohormonal influences. Inhibition of the renin-angiotensin system by angiotensin-converting enzyme inhibitors effectively reduces left ventricular hypertrophy and decreases morbidity and mortality due to heart failure, as well as slowing the progression of renal disease. Initial data from studies of angiotensin II type 1 (AT1) receptor blockers indicate that these agents should also be effective in reducing cardiac and renal damage. In this class, candesartan, by virtue of its tight and long-lasting binding to the AT1-receptor, provides pronounced 24-h blood pressure control with effective blockade of all the major negative cardiovascular effects of angiotensin II. Candesartan cilexetil has also been shown to be effective and well tolerated in combination with hydrochlorothiazide in those hypertensive patients who require more than one agent to reach their target blood pressure. PMID- 11059635 TI - Preserving target-organ function with candesartan cilexetil in patients with hypertension. AB - Epidemiological evidence suggests that reducing blood pressure alone in hypertensive patients delays the onset of cardiovascular events without necessarily preventing the progression of chronic target-organ disease, such as end-stage renal failure and heart failure. Successful clinical management of hypertensive patients will therefore not be possible unless therapies are aimed both at the effective control of blood pressure and at the preservation of target organ function. The new angiotensin II type I (AT1) receptor blocker candesartan cilexetil has been shown to be effective in reducing target-organ damage in animal models of hypertension, even at doses that do not produce significant reductions in blood pressure. Protective effects of candesartan cilexetil towards the heart and kidney have also been demonstrated in the clinical studies that have been conducted to date. Thus, candesartan cilexetil has been shown to induce regression of left ventricular hypertrophy within 8-12 weeks of treatment and to improve renal haemodynamics, both acutely and after 6 weeks of treatment in hypertensive patients. Furthermore, in hypertensive patients with co-existent non insulin-dependent diabetes mellitus and microalbuminuria, 12 weeks of treatment with candesartan cilexetil, 8-16 mg, significantly reduced urinary albumin excretion. Clinical evidence is therefore accumulating that the antihypertensive efficacy and tolerability profile already established for candesartan cilexetil is combined with the renal and cardioprotective effects necessary for optimal management of hypertension. PMID- 11059636 TI - Reducing cardiovascular morbidity and mortality in the elderly. AB - Candesartan cilexetil is highly effective at lowering blood pressure, whilst maintaining placebo-like tolerability, in a wide range of patient groups. Although the benefit of lowering blood pressure in elderly patients with moderate hypertension has been demonstrated in several large-scale clinical trials, elderly patients with mild hypertension have rarely been studied. The high incidence of cardiovascular and cerebrovascular mortality and morbidity, including dementia, in the elderly means that control of blood pressure is particularly important in this patient group. A major new international clinical trial - SCOPE (Study on COgnition and Prognosis in the Elderly) - has therefore been initiated. This is a prospective, randomized, double-blind, parallel comparison of the effects of candesartan cilexetil, 8 or 16 mg once daily, and placebo in about 5000 patients who will be followed for a mean of 2.5 years. SCOPE is the first study designed to assess the effect of antihypertensive therapy in elderly patients (70-89 years of age) with mild hypertension (sitting systolic blood pressure of 160-179 mmHg and/or sitting diastolic blood pressure of 90-99 mmHg). The primary objective of the study is to determine the effect of candesartan cilexetil on major cardiovascular events (cardiovascular death, non fatal stroke and myocardial infarction, and silent myocardial infarction), while an important secondary objective is to determine the effect of such treatment on the prevention of cognitive impairment. SCOPE should provide definitive evidence of the cardiovascular and cerebrovascular benefits of treating mildly hypertensive elderly patients with angiotensin II type 1 receptor blockers, which not only reduce blood pressure, but may also provide significant protection from the negative effects of angiotensin II on target organs. PMID- 11059637 TI - Exploring new treatment strategies in heart failure. AB - Heart failure remains a major and increasing cause of mortality and morbidity, even when the best available treatments are used. One of its key causes is neuroendocrine activation via the sympathetic nervous system and the renin angiotensin system (RAS). Neuroendocrine blockers of the sympathetic nervous system (beta-blockers) and of the RAS (angiotensin-converting enzyme [ACE] inhibitors and angiotensin II type 1 [AT1] receptor blockers) therefore have an important potential therapeutic role in heart failure. The promising results from clinical trials with beta-blockers suggest that these drugs will become an established part of the future management of patients with mild to moderate symptomatic heart failure. Blockade of the RAS with ACE inhibitors has also been shown to be effective in reducing the risk of morbidity and mortality in patients with heart failure. Blockade of the AT1-receptor, with agents such as candesartan, produces more specific and, theoretically, more complete blockade of the major negative cardiovascular effects of angiotensin II than is possible using ACE inhibitors, whilst maintaining placebo-like tolerability. Furthermore, AT1-receptor blockade leads to increased stimulation of the angiotensin II type 2 (AT2) receptor, which, according to experimental data, may have favourable cardiovascular effects. Following encouraging results from two pilot studies, a major new international study programme - CHARM (Candesartan in Heart failure - Assessment of Reduction in Mortality and morbidity) - has been initiated to define the clinical benefits of candesartan cilexetil in a wide variety of patients with symptomatic heart failure. CHARM is the first study to accept all relevant heart failure patients who may benefit from RAS blockade, irrespective of their left ventricular function or tolerance of ACE inhibitors. The 6500 patients to be recruited will be divided among three integrated outcome studies. Two of these studies will examine the effect of candesartan cilexetil versus placebo in patients with an ejection fraction of 40% or less who are tolerant or intolerant of ACE inhibitors. The third study arm will examine the benefits of candesartan cilexetil in a previously seldom studied group: those with symptomatic heart failure, but with preserved left-ventricular systolic function. Recruitment of patients into the study has started. PMID- 11059638 TI - Differential effects of CD40 stimulation on normal and neoplastic cell growth. AB - CD40 is a molecule in the tumor necrosis factor receptor/nerve growth factor receptor (TNFR/NGFR) family that is present on both normal and neoplastic B lineage cells. It is also expressed on carcinoma and melanoma cells and can be augmented with interferon gamma. CD40 stimulation in normal B cells has been demonstrated to promote normal B cell differentiation and growth in vitro. In contrast to these effects, CD40 stimulation by either anti-CD40 antibodies or a recombinant soluble CD40 ligand can inhibit the growth of human breast carcinomas and aggressive histology B lymphomas in vitro and in vivo. This is believed to occur by activation-induced cell death (AICD) in which stimuli that promote the growth of normal cell types inhibit the growth of neoplastic counterparts. This occurs through the induction of apoptosis, necrosis and/or cell cycle arrest. Thus, CD40 stimulation may be of potential clinical use in the treatment of carcinomas and B cell lymphomas. This review shall provide an overview of the various effects of CD40 stimulation on both normal and neoplastic cell types. PMID- 11059639 TI - Rethinking globally relevant vaccine strategies to human immunodeficiency virus type-1. AB - According to the latest UNAIDS figures for 1999 there were an estimated 30.6 million people living with HIV-1, with 16,000 new HIV infections per day. The only global strategy of combating new HIV infections is to make a vaccine that is affordable to developing countries, where greater than 90% of new infections occur, and that has enough efficacy to interrupt high rates of transmission. This review critically examines: 1) important immune parameters that should be considered which will allow an understanding of preventative vaccine design and 2) the mechanisms underlying immune destruction during HIV-1 infection that will facilitate design of therapeutic vaccines. A realistic goal of a preventative vaccine is to elicit protective immune responses in vaccinees that would prevent HIV-1 from replicating extensively in the host. Components of protective immunity are thought to include neutralizing antibodies (NAB) and cytotoxic T lymphocytes (CTL). Rethinking vaccine strategies has to take into account that HIV-1 vaccines must elicit primary cellular and humoral immunity via dendritic cell and Langerhan cell priming. It is only under these conditions that boosting immunity with subsequent vaccinations will allow high enough CTL effector cells and NAB titres to impede or to prevent HIV-1 replication. Success of therapeutic vaccine strategies, has to take into consideration the pathology of persistent immune stimulation by chronic HIV-1 infection. To re-stimulate immunity and re-direct immune responses, chronic immune stimulation by HIV-1 has to be alleviated by reducing high levels of viral antigen presentation by suppressing virus with antiretroviral agents. Such treatment courses may only have to be transient, long enough for immunity to respond to an immunogenic stimulus. Short-course drug therapy may then be an affordable option for many countries already carrying a high burden of HIV-1/AIDS. PMID- 11059640 TI - The immunological effects of interleukin 2 therapy in HIV+ patients. AB - Human immunodeficiency virus (HIV) infection produces a profound impairment of immune functions that antiretroviral therapy is unable to restore. Because of its immuno-enhancing properties, interleukin 2 (IL-2) has been used as a therapeutic tool in HIV+ subjects. IL-2 produces an increase of CD4 and CD8 lymphocyte absolute counts that is preferentially due to the expansion of the "naive" cells. In addition, IL-2 increases cytokine production from the cells of the immune system and is able to up-regulate the expression of cytokine receptors, such as the chemokine receptors CCR-5 and CXCR-4. Less informations on the IL-2 activity on the CD8 subset are available at the moment. The advent of highly active antiretroviral therapy has changed this scenario, making the IL-2 effects less clear-cut than previously hypothesized. We suggest that the ongoing studies will define the precise role of IL-2 in the therapy of HIV infection. PMID- 11059641 TI - Cellular immune activation, neopterin production, tryptophan degradation and the development of immunodeficiency. AB - Cellular (Th1-type) immune response is centrally involved in the pathogenesis of various diseases. Within the immunological cascades of Th1-type immunity, interferon gamma (IFN-gamma), among other cytokines, is critically involved. It triggers a series of immune-relevant reactions mostly directed towards forward regulation of the antigen specific immune response. However, in chronic states of immune activation, systemically increased IFN-gamma is no longer antigen specific and is associated with the development of immunodeficiency. IFN-gamma also stimulates the production of neopterin, a low-mass compound, in human monocytes/macrophages. Accordingly, neopterin concentrations in humans reflect the degree of Th1-type immune activation. Since IFN-gamma also stimulates the release of reactive oxygen species (ROS) from immunocompetents cells, the amount of neopterin produced also serves as an indirect estimate of oxidative stress. In parallel, IFN-gamma activates the degradation of tryptophan, which appears to limit the growth of intracellular pathogens and the proliferation of cells, including T lymphocytes. Thus, during persisting states of immune activation, the production of IFN-gamma is not only associated with forward regulation of the immune response, but also with immunosuppressive mechanisms. The increased formation of neopterin and degradation of tryptophan may result in a decreased T cell responsiveness and development of immunodeficiency. PMID- 11059642 TI - Role of immune-derived diffusible mediators in AIDS-associated neurological disorders. AB - Neurologic abnormalities are common in HIV-1 infected patients and often represent the dominant clinical manifestation of pediatric AIDS. Although the neurological dysfunction has been directly related to CNS invasion by HIV-1, the pathogenesis of neurologic disorders remains unclear. This review will first discuss the spectrum of potential interactions between HIV-1 and neural (neuronal and glial) cells, in the face of experimental data. Next, we will focus on the role of immune-derived cytokines and other soluble compounds which have been proposed to act as neurotoxic mediators and appear to play a role in the pathogenesis of AIDS-associated neurodegeneration. PMID- 11059643 TI - B cell development and primary immunodeficiencies with hypogammaglobulinemia. AB - The differentiation of B cells along the pathway of B cell development has been well characterized. In the bone marrow, the differentiation from pro-B cells to immature B cells can be defined by several surface antigens, such as a surrogate light chain. Immature B cells become mature B cells and then circulate in the peripheral blood as naive B cells. In the peripheral lymphoid tissues, naive B cells differentiate into memory B cells, which express the CD27 molecule, or plasma cells. Primary immunodeficiencies with hypogammaglobulinemia are caused by defects of the specific molecules which are needed for the B cell differentiation. Recent studies of the genes responsible for such immunodeficiencies have clarified B cell development as well as their pathogenesis. We discuss here the molecules affecting the B cell development and primary immunodeficiencies with hypogammaglobulinemia. PMID- 11059644 TI - Macrophages and HIV-1-associated dementia. AB - One of the strongest predictors for HIV-1-associated dementia is the presence of monocytic infiltration in perivascular areas of the brain. Therefore, macrophages have been suggested to play a major role in the development of this disease. This review focuses on possible mechanisms through which the macrophage may enhance disease progression by mediating neuronal damage. PMID- 11059645 TI - Ki-67 reactivity in primary fallopian tube cancers. AB - Forty-four cases of primary cancer of the fallopian tube (PFTC) were analyzed as to Ki-67 expression, grade, stage and the cancer histological type. Among patients with an average age of 57.5 years (range 38-70 years), 27 patients were FIGO I, 7 were FIGO II and 10 were FIGO III. Histological classification of PFTC revealed 18 cases of endometrioid type, 9 serous, 7 undifferentiated, 6 urothelial, 2 clear-cell and 2 of other type. Histological grading revealed 11 cases of G1, 16 of G2 and 17 of G3 tumors. The quantity of Ki-67 positive cells was counted on 300 cancer cells in random high-power fields (10 x 40) and recorded as the labeling index (LI, %). Positive staining for Ki-67 was shown in the nuclei in all cases. Ki-67 LI values ranged from 14.2 to 97.2% (median 36.1). Ki-67 LI values were graded as > or = 36.1% as high and <36.1% as low. We did not find any significant differences in Ki-67 LI values among tumors of various clinical stages, histological grades and histological types. The p value was statistically significant only for stage as a prognostic factor. PMID- 11059646 TI - Effect of recombinant IFN-gamma on igE-dependent leukotriene generation by peripheral blood leukocytes in patients with pollinosis and asthma. AB - Interferon gamma (IFN-gamma) is considered one of the causative and intensifying factors in inflammation. The reaction to allergens releases IFN-gamma, an immunomodulatory cytokine known to inhibit IgE synthesis and Th cell proliferation. The aim of the study was to evaluate the influence of IFN-gamma on leukotriene (LT) release in vitro, from human leukocytes of atopic patients with pollinosis and asthma. Thirty-eight patients were enrolled in the study: 15 with pollinosis and 23 asthmatics. In the presence of IL-3, leukocytes were stimulated with specific allergens. Other samples of leukocytes were preincubated with different concentrations of IFN-gamma for 15 min before allergen stimulation. The concentration of LT in supernatants was measured according to the CAST-ELISA procedure. We stated that IFN-gamma had significantly diminished LT release in a dose-dependent mode from the leukocytes of pollinotics. IFN-gamma did not change LT release in the asthmatic group, although, in leukocytes the small and medium basic production of LT, IFN-gamma caused a statistically significant fall in LT generation. PMID- 11059647 TI - The influence of amifostine used alone or in combination with 2 chlorodeoxyadenosine on normal and chronic myelogenous leukemia granulocyte macrophage progenitor cells in vitro. AB - We evaluated the influence of amifostine used alone or in combination with 2 chlorodeoxyadenosine (2-CdA) on the colony growth of normal and chronic myeloid leukemia (CML) granulocyte-macrophage progenitor cells (CFU-GM) in semisolid culture in vitro. Amifostine at a concentration of 1 mg/ml was either added directly to the culture medium of normal and CML CFU-GM, or mononuclear cells (MNCs) were first preincubated with amifostine at the same concentration, washed in Iscove's modified Dulbecco minimum essential medium (IDMEM) and then added to the culture medium. Amifostine used alone inhibited the growth of CML CFU-GM colonies to a higher degree than those of normal CFU-GM, but the differences were not statistically significant. Amifostine preincubated with MNCs and used together with the highest concentration of 2-CdA significantly inhibited the colony growth of CML CFU-GM as compared to 2-CdA alone (p<0.05). In contrast, the colony growth inhibition of normal CFU-GM was not significantly lower compared to 2-CdA used alone. Our studies suggest that 2-CdA used together with amifostine is more toxic to leukemic CFU-GM than to their normal counterparts. PMID- 11059648 TI - Serum levels of cytokines in alcoholic liver cirrhosis and pancreatitis. AB - Although altered cytokine homeostasis has been implicated in the pathogenesis of both alcoholic liver and pancreas diseases, the serum cytokine pattern characteristic of concomitant alcoholic liver cirrhosis and pancreatitis has not been examined. In this paper we examine the serum levels of proinflammatory cytokines, such as IL-6, IL-8, TNF-alpha, and also antiinflammatory ones, such as IL-10 and TGF-beta, in 22 patients with alcoholic liver cirrhosis and 28 patients with chronic pancreatitis and compare them with those detected in the sera of 14 patients with concomitant alcoholic cirrhosis and pancreatitis. All patients were heavy alcohol drinkers, consuming more than 70 g of pure alcohol per day for at least 5 years. The control group consisted of 33 age- and sex-matched healthy subjects receiving an annual health examination. They were not addicted to alcohol and confirmed to be free of major cardiopulmonary, gastrointestinal and hepatobiliary-pancreatic diseases. The results indicated that the cytokine pattern in the sera of patients with concomitant liver cirrhosis and pancreatitis was characterized by increased levels of two proinflammatory cytokines: TNF alpha, the concentration of which seemed to be influenced by both liver and pancreas injury, and IL-6, which seemed to be rather connected with pancreas injury. Increased levels of IL-8, which were detected in the sera of patients with cirrhosis, pancreatitis and concomitant cirrhosis and pancreatitis, were rather connected with exacerbation of the disease processes which occurred only in some of the patients. No significant changes in the levels of IL-10 or TGF beta were detected in the sera of patients with chronic pancreatitis and concomitant cirrhosis and pancreatitis, while in patients with cirrhosis significantly decreased levels of IL-10 were found. A significant imbalance between proinflammatory/antiinflammatory signals was especially characteristic of alcoholic cirrhosis and concomitant cirrhosis with pancreatitis. PMID- 11059649 TI - Screening for germline p53 mutations in pediatric and adult patients of high-risk groups in Poland. AB - Germline mutations of the p53 gene lead to cell transformation in various tissues. Such a complex cancer phenotype makes it difficult to recognize the carriers of the defective allele. Several studies undertaken to identify high risk groups found germline p53 mutations in familial cancer aggregations and in patients with multiple tumors. We screened 189 pediatric and 48 adult patients. The high-risk groups comprised 41 patients with a family history of cancer and 35 with multiple neoplasms. Furthermore, 124 tumors were screened for somatic mutations. p53 exons 2 to 11 were analyzed by polymerase chain reaction and single strand conformation polymorphism (PCR-SSCP) followed by direct sequencing of abnormal DNA fragments. No germline p53 mutations were found and somatic mutations were detected in 5 of 59 sarcomas, globally, in 8 of 124 tumors. In conclusion, in Poland, p53 alterations do not seem very important for the predisposition to malignancy and development of sarcomas. PMID- 11059650 TI - Epstein-Barr virus association in pediatric abdominal non-Hodgkin-lymphomas from Turkey. AB - Recent studies have shown marked geographic variation associated with Epstein Barr virus (EBV) in pediatric Burkitt's lymphomas and Hodgkin's disease. In the present study we investigated 30 cases of pediatric extranodal high grade non Hodgkin's lymphomas (NHL) from Turkey with an abdominal localisation. To classify them histologically and to determine the role of EBV in these lymphomas, immunohistochemistry (IHC), in situ hybridisation (ISH) and polymerase chain reaction (PCR) were used. Our series contained two histologic types: the Burkitt's or Burkitt's-like lymphomas (BL/BLL) and high grade NHL. They all were of the B cell type. The immunoglobulin heavy chain gene rearrangement revealed monoclonality in 87% of the BL/BLL cases, in contrast to the NHL cases, showing monoclonality in only 43% of the cases. EBV was found in tumor cells in a high frequency, independent of the histological subtype. EBV strains A and B were detected in 9 cases, with a preponderance of the B subtype (4/9 BL/BLL; 4/9 NHL). Our data suggest that high grade NHLs with abdominal localisation of Turkish children show the pattern of immunodeficient lymphomas to some extent. PMID- 11059651 TI - Neurosurgery: a historical prologue to the future. 2000 Presidential address. AB - The author provides a brief history of the genesis of organized neurosurgery and, in particular, the formation and evolution of the American Association of Neurological Surgeons. The legacy of neurosurgery is noted and the author discusses the present and future practice of neurosurgery. PMID- 11059652 TI - Radiosurgery for Cushing's disease after failed transsphenoidal surgery. AB - OBJECT: Although transsphenoidal surgery has become the standard of care for Cushing's disease, it is often unsuccessful in normalizing cortisol production. In this study the authors investigate the safety and efficacy of gamma knife radiosurgery (GKRS) for Cushing's disease after failed transsphenoidal surgery. METHODS: The records of all patients who underwent GKRS at the authors' institution after unsuccessful transsphenoidal surgery for Cushing's disease were retrospectively reviewed. Successful treatment was considered a normal or below normal 24-hour urinary free cortisol (UFC) level. Records were also evaluated for relapse, new-onset endocrine deficiencies, interval change in tumor size, and visual complications. Forty-three patients underwent 44 gamma knife procedures with follow up ranging from 18 to 113 months (mean 39.1, median 44 months). Normal 24-hour UFC levels were achieved in 27 patients (63%) at an average time from treatment of 12.1 months (range 3-48 months). Three patients had a recurrence of Cushing's disease at 19, 37, and 38 months, respectively, after radiosurgery. New endocrine deficiencies were noted in seven patients (16%). Follow-up magnetic resonance images obtained in 33 patients revealed a decrease in tumor size in 24, no change in nine, and an increase in size in none of the patients. One patient developed a quadrantanopsia 14 months after radiosurgery despite having received a dose of only 0.7 Gy to the optic tract. CONCLUSIONS: Gamma knife radiosurgery appears to be safe and effective for the treatment of Cushing's disease refractory to pituitary surgery. Delayed recurrences and new hormone deficiencies may occur, indicating the necessity for regular long-term follow up. PMID- 11059653 TI - Hypopituitarism following traumatic brain injury and aneurysmal subarachnoid hemorrhage: a preliminary report. AB - OBJECT: Recognition of pituitary hormonal insufficiencies after head injury and aneurysmal subarachnoid hemorrhage (SAH) may be important, especially given that hypopituitarism-related neurobehavioral problems are typically alleviated by hormone replacement. In this prospective study the authors sought to determine the rate and risk factors of pituitary dysfunction after head injury and SAH in patients at least 3 months after insult. METHODS: Patients underwent dynamic anterior and posterior pituitary function testing. Results of the tests were compared with those of 18 age-, sex-, and body mass index-matched healthy volunteers. The 22 head-injured patients included 18 men and four women (mean age 28+/-10 years at the time of injury) with initial Glasgow Coma Scale (GCS) scores of 3 to 15. Eight patients (36.4%) had a subnormal response in at least one hormonal axis. Four were growth hormone (GH) deficient. Five patients (four men, all with normal testosterone levels, and one woman with a low estradiol level) exhibited an inadequate gonadotroph response. One patient had both GH and thyrotroph deficiency and another had both GH deficiency and borderline cortisol deficiency. At the time of injury, all eight patients with pituitary dysfunction had an initial GCS score of 10 or less and, compared with the 14 patients without dysfunction, were more likely to have had diffuse swelling, seen on initial computerized tomography scans (p < 0.05), and to have sustained a hypotensive or hypoxic insult (p = 0.07). Of two patients with SAH who were studied (Hunt and Hess Grade IV) both had GH deficiency. CONCLUSIONS: From this preliminary study, some degree of hypopituitarism appears to occur in approximately 40% of patients with moderate or severe head injury, with GH and gonadotroph deficiencies being most common. A high degree of injury severity and secondary cerebral insults are likely risk factors for hypopituitarism. Pituitary dysfunction also occurs in patients with poor-grade aneurysms. Postacute pituitary function testing may be warranted in most patients with moderate or severe head injury, particularly those with diffuse brain swelling and those sustaining hypotensive or hypoxic insults. The neurobehavioral effects of GH replacement in patients suffering from head injury or SAH warrant further study. PMID- 11059654 TI - Multiple pituitary adenomas in Cushing's disease. AB - OBJECT: Clinically evident multiple pituitary adenomas rarely occur. The authors assess the incidence and clinical relevance of multiple adenomas in Cushing's disease. METHODS: A prospective clinical database of 660 pituitary surgeries was analyzed to assess the incidence of multiple pituitary adenomas in Cushing's disease. Relevant radiographic scans, medical records, and histopathological reports were reviewed. Thirteen patients with at least two separate histopathologically confirmed pituitary adenomas were identified. Prolactinomas (nine patients) were the most common incidental tumors. Other incidental tumors included secretors of growth hormone ([GH], one patient) and GH and prolactin (two patients), and a null-cell tumor (one patient). In two patients, early repeated surgery was performed because the initial operation failed to correct hypercortisolism, in one instance because the tumor excised at the initial surgery was a prolactinoma, not an adrenocorticotropic hormone-secreting tumor. One patient had three distinct tumors. CONCLUSIONS: Multiple pituitary adenomas are rare, but may complicate management of patients with pituitary disease. PMID- 11059655 TI - Sellar reconstruction with resorbable vicryl patches, gelatin foam, and fibrin glue in transsphenoidal surgery: a 10-year experience with 376 patients. AB - OBJECT: Closure of the sella turcica after transsphenoidal surgery is mainly accomplished with autologous muscle fascia and fat or muscle; this requires a second surgical incision. The authors review the results of using resorbable vicryl patches, gelatin foam, and fibrin glue for sellar reconstruction. METHODS: A review was conducted of 376 consecutive patients who underwent surgery for pituitary adenomas, cysts, or subdiaphragmatic craniopharyngiomas in the sella turcica that the senior author (R.W.S.) had performed or directly supervised over the last 10 years. The sellar reconstruction was performed with a commercially available, synthetic absorbable patch composed of polyglactin 910/poly-p dioxanone, gelatin foam, and fibrin glue. The patch is essentially resorbed in 2 to 3 months and replaced by fibrous collagen tissue. There were 117 small, 112 medium-sized, and 147 large lesions. The overall nonendocrine postoperative morbidity rate was 2.8%, and included visual deterioration, meningitis, secondary epistaxis, nasal septum complication, and cerebrospinal fluid (CSF) leakage. Two patients with macroadenomas needed reoperation for persistent CSF leakage, which comprised 0.5% of the whole series or 0.8% of the 259 patients with medium-sized or large lesions. There was no mortality and no morbidity related to the implanted material, and in particular no delayed empty sella syndrome. CONCLUSIONS: Closure of the sella turcica with a synthetic absorbable vicryl patch, gelatin foam, and fibrin glue after transsphenoidal surgery is safe and very effective in preventing postoperative CSF fistulas. The use of this technique obviates the need for a second surgical incision and shortens the operating time. Because of the progressive resorption of the substitute material, the interpretation of postoperative magnetic resonance studies was not significantly hindered. PMID- 11059656 TI - Surgical treatment of intractable epilepsy accompanying cortical dysplasia. AB - OBJECT: Surgical treatment of cortical dysplasia (CD) together with intractable seizures is challenging because both visualization and localization of the lesion are difficult, correlation with seizure foci requires comprehensive study, and the surgical outcomes reported thus far are unsatisfactory. The authors report their experience in the surgical treatment of CD classified according to a surgical point of view. METHODS: The definition of CD used in this study was a dysplastic lesion visible on magnetic resonance (MR) images or a lesion that, although not visible on MR images, was diagnosed as moderate-to-severe dysplasia by using pathological analysis. During the last 4.5 years, the authors treated 36 patients with intractable epilepsy accompanied by CD. They divided the 36 cases of CD into four characteristic groups: Group A, diffuse bilateral hemispheric dysplasia; Group B, diffuse lobar dysplasia; Group C, focal dysplasia; and Group D, a moderate to severe degree of CD with a normal appearance on MR images. All but one patient in Group C were monitored in the epilepsy monitoring unit by using subdural electrodes for seizure localization and functional mapping. The incidence of CD among a cohort of 291 patients who had undergone epilepsy surgery at the authors' center during the study period was 12.4%. The mean age of the 36 patients was 21.3 years and the mean age at seizure onset was 8.5 years. The mean follow-up period was 26 months. Twenty-six patients (72.2%) belonged to Engel Class I or II (20 and six, respectively). There were five cases in Group A, nine in Group B, nine in Group C, and 13 in Group D. Patients in Groups A and B were significantly younger at seizure onset and had significantly poorer surgical outcomes compared with patients in Groups C and D (p < 0.05). If outcome is compared on the basis of the extent of removal of CD, patients in whom CD was completely removed had significantly better outcomes than those in whom CD was only partially removed (p < 0.001). CONCLUSIONS: The authors conclude that intractable epilepsy accompanied by CD can be treated surgically using comprehensive preoperative approaches. Deliberate resective procedures aimed at complete removal of dysplastic tissue ensure excellent seizure control without permanent neurological deficit. PMID- 11059657 TI - Functional magnetic resonance imaging of somatosensory cortex activity produced by electrical stimulation of the median nerve or tactile stimulation of the index finger. AB - OBJECT: Functional magnetic resonance (fMR) imaging was used to determine patterns of cerebral blood flow changes in the somatosensory cortex that result from median nerve stimulation (MNS). METHODS: Ten healthy volunteers underwent stimulation of the right median nerve at frequencies of 5.1 Hz (five volunteers) and 50 Hz (five volunteers). The left median nerve was stimulated at frequencies of 5.1 Hz (two volunteers) and 50 Hz (five volunteers). Tactile stimulation (with a soft brush) of the right index finger was also applied (three volunteers). Functional MR imaging data were transformed into Talairach space coordinates and averaged by group. Results showed significant activation (p < 0.001) in the following regions: primary sensorimotor cortex (SMI), secondary somatosensory cortex (SII), parietal operculum, insula, frontal cortex, supplementary motor area, and posterior parietal cortices (Brodmann's Areas 7 and 40). Further analysis revealed no statistically significant difference (p > 0.05) between volumes of cortical activation in the SMI or SII resulting from electrical stimuli at 5.1 Hz and 50 Hz. There existed no significant differences (p > 0.05) in cortical activity in either the SMI or SII resulting from either left- or right-sided MNS. With the exception of the frontal cortex, areas of cortical activity in response to tactile stimulation were anatomically identical to those regions activated by electrical stimulation. In the SMI and SII, activation resulting from tactile stimulation was not significantly different (p > 0.05) from that resulting from electrical stimulation. CONCLUSIONS: Electrical stimulation of the median nerve is a reproducible and effective means of activating multiple somatosensory cortical areas, and fMR imaging can be used to investigate the complex network that exists between these areas. PMID- 11059658 TI - Magnetic resonance imaging stereotactic target localization for deep brain stimulation in dystonic children. AB - OBJECT: The actual distortion present in a given series of magnetic resonance (MR) images is difficult to establish. The purpose of this study was to validate an MR imaging-based methodology for stereotactic targeting of the internal globus pallidus during electrode implantation in children in whom general anesthesia had been induced. METHODS: Twelve children (mean follow up 1 year) suffering from generalized dystonia were treated with deep brain stimulation by using a head frame and MR imaging. To analyze the influence of distortions at every step of the procedure, the geometrical characteristics of the frame were first controlled using the localizer as a phantom. Then pre- and postoperative coordinates of fixed anatomical landmarks and electrode positions, both determined with the head frame in place, were statistically compared. No significant difference was observed between theoretical and measured dimensions of the localizer (Student's t-test, ?t? > 2.2 for 12 patients) in the x, y, and z directions. No significant differences were observed (Wilcoxon paired-sample test) between the following: 1) pre- and postoperative coordinates of the anterior commissure (AC) (deltax = 0.3+/-0.29 mm and deltay = 0.34+/-0.32 mm) and posterior commissure (PC) (deltax = 0.15+/-0.18 mm and deltay = 0.34+/-0.25 mm); 2) pre- and postoperative AC-PC distance (deltaL = 0.33+/-0.22 mm); and 3) preoperative target and final electrode position coordinates (deltax = 0.24+/-0.22 mm; deltay = 0.19+/-0.16 mm). CONCLUSIONS: In the authors' center, MR imaging distortions did not induce detectable errors during stereotactic surgery in dystonic children. Target localization and electrode implantation could be achieved using MR imaging alone after induction of general anesthesia. The remarkable postoperative improvement in these patients confirmed the accuracy of the procedure (Burke-Marsden-Fahn Dystonia Rating Scale score delta = -83.8%). PMID- 11059659 TI - Relationship between drainage catheter location and postoperative recurrence of chronic subdural hematoma after burr-hole irrigation and closed-system drainage. AB - OBJECT: This study was conducted to determine the best position for the subdural drainage catheter to achieve a low recurrence rate after burr-hole irrigation and closed-system drainage of chronic subdural hematoma (CSDH). METHODS: The authors studied 63 patients with CSDH in whom the drainage catheter tip was randomly placed and precisely determined on postoperative computerized tomography (CT) scans and 104 patients with CSDH in whom CT scans were obtained 7 days postsurgery. The location of the subdural drainage catheter, the maximum postoperative width of the subdural space, and the percentage of the ipsilateral subdural space occupied by air postoperatively were determined and compared with the postoperative recurrence and reoperation rates. Patients with parietal or occipital drainage had a higher rate of CSDH recurrence and much more subdural air than those with frontal drainage. In addition, patients with residual subdural air demonstrated on CT scans obtained 7 days postsurgery also had a higher recurrence rate than those without subdural air collections. Furthermore, patients with a subdural space wider than 10 mm on CT scans obtained 7 days postsurgery had a higher recurrence rate than those with a space measuring 10 mm or less. CONCLUSIONS: The incidence of postoperative fluid reaccumulation seems to be reduced by placing the tip of the drainage catheter in the frontal convexity and by removing subdural air during or after surgery. PMID- 11059660 TI - Chronic subdural hematoma: evaluation of the clinical significance of postoperative drainage volume. AB - OBJECT: A wide variation in postoperative drainage volumes is observed during treatment of chronic subdural hematoma (CSDH) with twist-drill or burr-hole craniostomy and closed-system drainage. In this study the authors investigate the causes of the variation, the clinical significance thereof, and its influence on treatment outcome. METHODS: A total of 175 cases were investigated between January 1991 and December 1997. Of these, 145 patients had surgery for CSDH, of whom 30 had bilateral lesions. The cases of CSDH were divided into five subtypes (low-density, isodense, high-density, mixed-density, and layering types) on the basis of the brain computerized tomography (CT) findings. Burr-hole craniostomies with closed-system drainage were performed in all patients and the drainage was maintained for 5 days, during which daily amounts of fluid were measured. The mean drainage volume over 5 days was 320 ml, with the largest volume (413 ml) seen in the low-density type and the smallest (151 ml) in the mixed-density type of CSDH. There were recurrences in six patients (seven instances, 4%). The mixed density type had the highest recurrence rate (8.6%), whereas there was no recurrence for the low-density type. There were no recurrences in 81 patients in whom the total drainage volumes for 5 days were more than 200 ml, but there were recurrences in six (seven instances) of 94 patients in whom the total drainage volume was less than 200 ml. CONCLUSIONS: The postoperative drainage volumes varied greatly because of differences in the outer membrane permeability of CSDH, and such variation seems to be related to the findings on the CT scans obtained preoperatively. Patients with CSDH in whom there is less postoperative drainage than expected should be carefully observed, with special attention paid to the possibility of recurrence. PMID- 11059661 TI - High level of extracellular potassium and its correlates after severe head injury: relationship to high intracranial pressure. AB - OBJECT: Disturbed ionic and neurotransmitter homeostasis are now recognized as probably the most important mechanisms contributing to the development of secondary brain swelling after traumatic brain injury (TBI). Evidence obtained in animal models indicates that posttraumatic neuronal excitation by excitatory amino acids leads to an increase in extracellular potassium, probably due to ion channel activation. The purpose of this study was therefore to measure dialysate potassium in severely head injured patients and to correlate these results with measurements of intracranial pressure (ICP), patient outcome, and levels of dialysate glutamate and lactate, and cerebral blood flow (CBF) to determine the role of ischemia in this posttraumatic ion dysfunction. METHODS: Eighty-five patients with severe TBI (Glasgow Coma Scale Score < 8) were treated according to an intensive ICP management-focused protocol. All patients underwent intracerebral microdialyis. Dialysate potassium levels were analyzed using flame photometry, and dialysate glutamate and dialysate lactate levels were measured using high-performance liquid chromatography and an enzyme-linked amperometric method in 72 and 84 patients, respectively. Cerebral blood flow studies (stable xenon computerized tomography scanning) were performed in 59 patients. In approximately 20% of the patients, dialysate potassium values were increased (dialysate potassium > 1.8 mM) for 3 hours or more. A mean amount of dialysate potassium greater than 2 mM throughout the entire monitoring period was associated with ICP above 30 mm Hg and fatal outcome, as were progressively rising levels of dialysate potassium. The presence of dialysate potassium correlated positively with dialysate glutamate (p < 0.0001) and lactate (p < 0.0001) levels. Dialysate potassium was significantly inversely correlated with reduced CBF (p = 0.019). CONCLUSIONS: Dialysate potassium was increased after TBI in 20% of measurements. High levels of dialysate potassium were associated with increased ICP and poor outcome. The simultaneous increase in dialysate potassium, together with dialysate glutamate and lactate, supports the concept that glutamate induces ionic flux and consequently increases ICP, which the authors speculate may be due to astrocytic swelling. Reduced CBF was also significantly correlated with increased levels of dialysate potassium. This may be due to either cell swelling or altered vasoreactivity in cerebral blood vessels caused by higher levels of potassium after trauma. Additional studies in which potassium sensitive microelectrodes are used are needed to validate these ionic events more clearly. PMID- 11059662 TI - Cerebral microdialysis monitoring: determination of normal and ischemic cerebral metabolisms in patients with aneurysmal subarachnoid hemorrhage. AB - OBJECT: The success of treatment for delayed cerebral ischemia is time dependent, and neuronal monitoring methods that can detect early subclinical levels of cerebral ischemia may improve overall treatment results. Cerebral microdialysis may represent such a method. The authors' goal was to characterize patterns of markers of energy metabolism (glucose, pyruvate, and lactate) and neuronal injury (glutamate and glycerol) in patients with subarachnoid hemorrhage (SAH), in whom ischemia was or was not suspected. METHODS: By using low-flow intracerebral microdialysis monitoring, central nervous system extracellular fluid concentrations of glucose, pyruvate, lactate, glutamate, and glycerol were determined in 46 patients suffering from poor-grade SAH. The results in two subgroups were analyzed: those patients with no clinical or radiological signs of cerebral ischemia (14 patients) and those who succumbed to brain death (five patients). Significantly lower levels of energy substrates and significantly higher levels of lactate and neuronal injury markers were observed in patients with severe and complete ischemia when compared with patients without symptoms of ischemia (glucose 0 compared with 2.12+/-0.15 mmol/L; pyruvate 0 compared with 151+/-11.5 micromol; lactate 6.57+/-1.07 compared with 3.06+/-0.32 mmol/L; glycerol 639+/-91 compared with 81.6+/-12.4 micromol; and glutamate 339+/-53.4 compared with 14+/-3.33 micromol). Immediately after catheter placement, glutamate concentrations declined over the first 4 to 6 hours to reach stable values. The remaining parameters exhibited stable values after 1 to 2 hours. CONCLUSIONS: The results confirm that intracerebral microdialysis monitoring of patients with SAH can be used to detect patterns of cerebral ischemia. The wide range from normal to severe ischemic values calls for additional studies to characterize further incomplete and possible subclinical levels of ischemia. PMID- 11059663 TI - Impaired cerebral mitochondrial function after traumatic brain injury in humans. AB - OBJECT: Oxygen supply to the brain is often insufficient after traumatic brain injury (TBI), and this results in decreased energy production (adenosine triphosphate [ATP]) with consequent neuronal cell death. It is obviously important to restore oxygen delivery after TBI; however, increasing oxygen delivery alone may not improve ATP production if the patient's mitochondria (the source of ATP) are impaired. Traumatic brain injury has been shown to impair mitochondrial function in animals; however, no human studies have been previously reported. METHODS: Using tissue fractionation procedures, living mitochondria derived from therapeutically removed brain tissue were analyzed in 16 patients with head injury (Glasgow Coma Scale Scores 3-14) and two patients without head injury. Results revealed that in head-injured patients mitochondrial function was impaired, with subsequent decreased ATP production. CONCLUSIONS: Decreased oxygen metabolism due to mitochondrial dysfunction must be taken into account when clinically defining ischemia and interpreting oxygen measurements such as jugular venous oxygen saturation, arteriovenous difference in oxygen content, direct tissue oxygen tension, and cerebral blood oxygen content determined using near infrared spectroscopy. Restoring mitochondrial function might be as important as maintaining oxygen delivery. PMID- 11059664 TI - Neurobehavioral protection by the neuronal calcium channel blocker ziconotide in a model of traumatic diffuse brain injury in rats. AB - OBJECT: Abnormal accumulation of intracellular calcium following traumatic brain injury (TBI) is thought to contribute to a cascade of cellular events that lead to neuropathological conditions. Therefore, the possibility that specific calcium channel antagonists might exert neuroprotective effects in TBI has been of interest. The focus of this study was to examine whether Ziconotide produces such neuroprotective effects. METHODS: The authors report that the acceleration deceleration model of TBI developed by Marmarou, et al., induces a long-lasting deficit of neuromotor and behavioral function. The voltage-sensitive calcium channel blocker Ziconotide (also known as SNX-111 and CI-1009) exerts neuroprotective effects in this model of diffuse brain injury (DBI) in rats. The dose and time of injection of Ziconotide chosen for the present study was based on the authors' previous biochemical studies of mitochondria. Rats were trained in a series of motor and memory tasks, following which they were subjected to DBI using the Marmarou, et al., model. At 3, 5, and 24 hours, all rats were injected with 2 mg/kg Ziconotide for a total cumulative dose of 6 mg/kg Ziconotide. Control brain-injured animals were injected with an equal volume of saline vehicle at each of these time points. The rats were tested for motor and cognitive performance at 1, 3, 7,14, 21, 28, 35, and 42 days postinjury. Saline treated rats displayed severe motor and cognitive deficits after DBI. Compared with saline-treated control animals, rats treated with Ziconotide displayed better motor performance during inclined plane, beam balance, and beam walk tests; improved memory while in the radial arm maze; and improved learning while in the Morris water maze. CONCLUSIONS: These results demonstrated that the acceleration-deceleration model, which had been developed by Marmarou, et al., induces severe motor and cognitive deficits. We also demonstrated that Ziconotide exhibits substantial neuroprotective activity in this model of TBI. Improvement was observed in both motor and cognitive tasks, even though treatment was not initiated until 3 hours after injury. These findings support the development of neuronal N-type calcium channel antagonists as useful therapeutic agents in the treatment of TBI. PMID- 11059665 TI - Improvement in mitochondrial dysfunction as a new surrogate efficiency measure for preclinical trials: dose-response and time-window profiles for administration of the calcium channel blocker Ziconotide in experimental brain injury. AB - OBJECT: Determining the efficacy of a drug used in experimental traumatic brain injury (TBI) requires the use of one or more outcome measures such as decreased mortality or fewer neurological and neuropsychological deficits. Unfortunately, outcomes in these test batteries have a fairly large variability, requiring relatively large sample sizes, and administration of the tests themselves is also very time consuming. The authors previously demonstrated that experimental TBI and human TBI induce mitochondrial dysfunction. Because mitochondrial dysfunction is easy to assess compared with neurobehavioral endpoints, it might prove useful as an outcome measure to establish therapeutic time windows and dose-response curves in preclinical drug testing. This idea was tested in a model of TBI in rats. METHODS: Animals treated with the selective N-type voltage-sensitive calcium channel blocker Ziconotide (also known as SNX-111 and CI-1009) after cortical impact displayed significant improvement in brain mitochondrial function. When a single intravenous bolus injection of 4 mg/kg Ziconotide was given at different time intervals, ranging from 15 minutes before injury to 10 hours after injury, mitochondrial function was improved at all time points, but more so between 2 and 6 hours postinjury. The authors evaluated the effects on mitochondrial function of Ziconotide at different doses by administering 0.5 to 6 mg/kg as a single bolus injection 4 hours after injury, and found 4 mg/kg to be the optimum dose. CONCLUSIONS: The authors established these time-window profiles and dose-response curves on the basis of mitochondrial outcome measures in a total of 42 rats because there were such low standard deviations in these tests. Establishing similar time-window profiles and dose-response curves by using neurobehavioral endpoints would have required using 114 rats in much more elaborate experiments. PMID- 11059666 TI - No additional neuroprotection provided by barbiturate-induced burst suppression under mild hypothermic conditions in rats subjected to reversible focal ischemia. AB - OBJECT: Mild-to-moderate hypothermia is increasingly used for neuroprotection in humans. However, it is unknown whether administration of barbiturate medications in burst-suppressive doses-the gold standard of neuroprotection during neurovascular procedures-provides an additional protective effect under hypothermic conditions. The authors conducted the present study to answer this question. METHODS: Thirty-two Sprague-Dawley rats were subjected to 90 minutes of middle cerebral artery occlusion and randomly assigned to one of four treatment groups: 1) normothermic controls; 2) methohexital treatment (burst suppression); 3) induction of mild hypothermia (33 degrees C); and 4) induction of mild hypothermia plus methohexital treatment (burst suppression). Local cerebral blood flow was continuously monitored using bilateral laser Doppler flowmetry and electroencephalography. Functional deficits were quantified and recorded during daily neurological examinations. Infarct volumes were assessed histologically after 7 days. Methohexital treatment, mild hypothermia, and mild hypothermia plus methohexital treatment reduced infarct volumes by 32%, 71%, and 66%, respectively, compared with normothermic controls. Furthermore, mild hypothermia therapy provided the best functional outcome, which was not improved by additional barbiturate therapy. CONCLUSIONS: The results of this study indicate that barbiturate-induced burst suppression is not required to achieve maximum neuroprotection under mild hypothermic conditions. The magnitude of protection afforded by barbiturates alone appears to be modest compared with that provided by mild hypothermia. PMID- 11059667 TI - Ischemic tolerance in the rat neocortex following hypothermic preconditioning. AB - OBJECT: Ischemic neuronal damage associated with neurological and other types of surgery can have severe consequences for functional recovery after surgery. Hypothermia administered during and/or after ischemia has proved to be clinically beneficial and its effects often rival or exceed those of other therapeutic strategies. In the present study the authors examined whether transient hypothermia is an effective preconditioning stimulus for inducing ischemic tolerance in the brain. METHODS: Adult rats were subjected to a 20-minute period of hypothermic preconditioning followed by an interval ranging from 6 hours to 7 days. At the end of this interval, the animals were subjected to transient focal ischemia induced by clamping one middle cerebral artery and both carotid arteries for 1 hour. The volume of cerebral infarction was assessed 1 or 7 days postischemia. In the first series of experiments, hypothermic preconditioning (28.5 degrees C) with a postconditioning interval of 1 day reduced the extent of cerebral infarction measured 1 and 7 days postischemia. In the second series, hypothermic preconditioning (31.5 degrees C) with postconditioning intervals of 6 hours, 1 day, or 2 days (but not 7 days) reduced the extent of cerebral infarction measured 1 day postischemia. Treatment with the protein synthesis inhibitor anisomycin blocked the protective effect of hypothermic preconditioning. In a final series of experiments, in vitro brain slices prepared from hypothermia-preconditioned (nonischemic) animals were shown to tolerate a hypoxic challenge better than slices prepared from unconditioned animals. CONCLUSIONS: These findings indicate that hypothermic preconditioning induces a form of delayed tolerance to focal ischemic damage. The time course over which tolerance occurs and the ability of a protein synthesis inhibitor to block tolerance suggest that increased expression of one or more gene products is necessary to establish tissue tolerance following hypothermia. The attenuation of hypoxic injury in vitro following in vivo preconditioning indicates that tolerance is due, at least in part, to direct effects on the brain neuropil. Hypothermic preconditioning could provide a relatively low-risk approach for improving surgical outcome after invasive surgery, including high-risk neurological and cardiovascular procedures. PMID- 11059668 TI - Long-term testing of an intracranial pressure monitoring device. AB - OBJECT: Long-term monitoring of intracranial pressure (ICP) is limited by the lack of an implantable sensor with low drift. The goal of this study was to demonstrate that a new capacitive transducer system will produce accurate and stable ICP records over extended periods. METHODS: Intracranial pressure sensors were implanted into the frontal white matter of four dogs. In addition, a fluid filled catheter was placed in the cisterna magna (CM) to measure cerebrospinal fluid (CSF) pressure. The animals were tested using standard physiological maneuvers such as jugular vein compression, head elevation, and CSF withdrawal from and saline injection into the CM to verify that the ICP sensor precisely matched CSF pressure changes. The mean ICP pressure and CM pressure were compared for months to demonstrate that the transducer system produced minimal drift over time. The change in the ICP sensor record closely duplicated that of the CSF waveform in the CM in response to well-known physiological stimuli. More important, mean ICP pressure remained within 3 mm Hg of CM pressure for months, with a mean difference of less than 0.3 mm Hg. Histological examination of the dog brains revealed only minimal tissue reaction to the presence of the sensor. CONCLUSIONS: The authors demonstrate a new implantable solid-state sensor that reliably measures ICP for months, with minimal drift. The clinical application of this sensor and its telemetry is for long-term monitoring of patients with head injury, mass lesions, and hydrocephalus. PMID- 11059669 TI - Arterial dissections of penetrating cerebral arteries causing hypertension induced cerebral hemorrhage. AB - OBJECT: For the past 130 years, it has been believed that hypertension-induced cerebral hemorrhages are the result of ruptures of microaneurysms or ruptures of arteries that have degenerative changes. The majority of previous investigations have focused on autopsied brain. In this study, the authors attempted to verify the cause of hypertension-induced cerebral hemorrhage by using surgical specimens of the penetrating arteries responsible for the hemorrhages. METHODS: Between 1997 and 1999, the authors performed pathological studies in surgical specimens of lenticulostriate arteries that had been confirmed during microsurgery to be the cause of hypertension-induced hemorrhage of the putamen. Nineteen lenticulostriate arteries were collected from 12 patients. Fifteen of these arteries were verified as the pathological causes of hemorrhage. They included six arterial dissections, six arterial ruptures with substantial degenerative changes, and three arterial ruptures with few degenerative changes. The pathological findings in the lenticulostriate artery dissections were similar to those of typical arterial dissections in major cerebral arteries. CONCLUSIONS: To the best of the authors' knowledge, arterial dissections of lenticulostriate arteries have not been identified as a cause of hypertension-induced cerebral hemorrhages. When penetrating arteries are included as causative vessels, cerebral arterial dissections may be much more common than previously thought. PMID- 11059670 TI - Aberrant p21 regulation in radioresistant primary glioblastoma multiforme cells bearing wild-type p53. AB - OBJECT: A clearer understanding of the cellular mechanisms involved in the response to ionizing radiation is pivotal to the development of new therapeutic strategies for glioblastoma multiforme (GBM). To gain insight into dynamic functional aspects of cell cycle regulation and the control of apoptosis in GBMs, the authors investigated the molecular changes induced by ionizing radiation in genetically characterized primary GBMs in vitro compared with secondary GBMs, Grades II and III gliomas, and three GBM cell lines. METHODS: Irradiation of primary GBMs bearing wild-type (wt) p53 invariably fails to invoke the G, checkpoint and apoptosis in vitro. In approximately half of these primary GBMs a defect lies at or above the level of p53 because transcriptional activation of p21 and bax after irradiation does not occur. The failure of a p21 response to irradiation is invariably accompanied by overexpression of p21 mRNA under nonirradiated conditions. In all remaining primary GBMs transcriptional activation of p21 after irradiation does occur, suggesting that a defect downstream from p21 prevents G, arrest. CONCLUSIONS: These results show that the G, checkpoint and the p53 pathway are dysfunctional in primary GBMs in vitro, despite the presence of an intact p53 gene. The data also suggest that primary GBMs may be divided into two categories on the basis of their p21 response to irradiation. PMID- 11059671 TI - Reversal of thalamic hand syndrome by long-term motor cortex stimulation. AB - The authors describe a case of complete recovery from the so-called "thalamic hand" syndrome following chronic motor cortex stimulation in a 64-year-old man suffering from poststroke thalamic central pain. As of the 2-year follow-up examination, the patient's dystonia and pain are still controlled by electrical stimulation. It is speculated that a common mechanism in which the thalamocortical circuit loops are rendered out of balance may sustain hand dystonia and central pain in this case of thalamic syndrome. To the authors' knowledge this is the first reported case of its kind. PMID- 11059672 TI - Cortical reorganization after digit-to-hand replantation. AB - Functional recovery after digit-to-hand replantation depends on the interaction of various factors. In addition to peripheral mechanisms, cortical and subcortical reorganization of digit representation may play a substantial role in the recovery process. However, cortical processes during the first months after replantation are not well understood. In this 25-year-old man who had traumatically lost digits II to V (DII-V) on his right hand, the authors used magnetoencephalographic source imaging to document the recovery of somatosensory cortical responses after tactile stimulation at four sites on the replanted digits. Successful replantation of DIV and DV was accomplished at the original position of DIII and DIV with mixed innervation. Cortical evoked fields could be recorded starting from the 10th week after digit-to-hand replantation. Initially, signals from all sites showed decreased amplitudes and prolonged latencies. In the subsequent six recordings obtained between the 12th and 55th week postreplantation, a continuous increase in amplitude but only a slight recovery of latencies were observed. Components of the recorded somatosensory evoked fields were localized in the primary somatosensory cortex (SI). The localizations of the replanted DIV showed a gradual lateral-inferior shift in the somatosensory cortex over time, indicating cortical reorganization caused by altered peripheral input. The authors infer from this shift that the original cortical area of the missing finger (DII) was taken over by the replanted finger. From these data the authors conclude that magnetic source imaging might be a reliable noninvasive method to evaluate surgical nerve repair and that cortical reorganization of SI is involved in the regeneration process following peripheral nerve injury. PMID- 11059673 TI - Spontaneous regression of a germinoma in the pineal body after placement of a ventriculoperitoneal shunt. AB - The authors report a case in which a germinoma in the pineal body displayed spontaneous regression after placement of a ventriculoperitoneal (VP) shunt. Spontaneous regression of malignant tumors is extremely rare, occurring in only one of 60,000 to 100,000 patients. Although in rare cases spontaneous regression is known to occur in patients with testicular seminomas, only one case of spontaneous regression of a primary pineal germinoma has so far been reported. In the present case a 17-year-old man presented with headache. A tumor in the pineal body and acute hydrocephalus were revealed by head computerized tomography (CT) and magnetic resonance (MR) imaging, and VP shunt placement was performed. Computerized tomography scanning of the head was performed four times during a 2 week period following the operation, and the patient was temporarily discharged to return to school. At the time of discharge, CT scanning demonstrated no change in the size of the tumor. Two months later, the patient was readmitted to the hospital to undergo surgery. At that time, head MR imaging revealed regression of the tumor. The pathological diagnosis of the lesion was germinoma. The patient underwent three courses of chemotherapy, during which carboplatin and etoposide were administered, in addition to a 24-Gy dose of radiotherapy. No manifestations of nerve impairment were noticed, and the patient was observed on an outpatient basis. The authors think that the factors involved in tumor regression included the effects of the VP shunt, the effects of radiation absorbed during head CT scanning, and the role of the patient's own immune response. However, no conclusion has been reached concerning the actual cause. PMID- 11059674 TI - Diffuse vertebral body metastasis from a glioblastoma multiforme: a technetium 99m Sestamibi single-photon emission computerized tomography study. AB - The authors report on a case of right temporal glioblastoma multiforme (GBM) that metastasized to multiple bone regions (dorsolumbar vertebrae and iliac bone) 8 months after initial diagnosis, despite combined radio- and chemotherapy. Results of a whole-bone single-photon emission computerized tomography (SPECT) study using the imaging agent Sestamibi (MIBI) revealed extracranial metastases from the GBM. A magnetic resonance imaging study of the dorsolumbar spinal region completed the radiological investigation. Cells immunoreactive to glial fibrillary acidic protein were observed in a specimen obtained from the right iliac bone. Postmortem examination confirmed metastasis to extracranial bone and revealed two other metastatic localizations in the lung and heart. This is the first reported case of extracranial bone metastasis from a GBM demonstrated on a whole-bone MIBI SPECT scan. In patients with malignant glioma and lower-back pain (especially prolonged pain), bone metastasis, although uncommon, does occasionally occur and its possibility should be investigated; a MIBI SPECT study may prove useful in this regard. PMID- 11059675 TI - Endoscopically guided fenestration of the choroidal fissure for treatment of trapped temporal horn. AB - In recent years endoscopic procedures have been used more frequently to treat loculated hydrocephalus. The trapped temporal horn, a specific type of loculation, has traditionally been treated by means of ventricular shunt placement. By opening up loculations, however, this procedure can be simplified or, in some cases, even avoided. In this report the author discusses a case of trapped temporal horn that was caused by fungal meningitis and treated using endoscopically guided fenestration of the choroidal fissure, leading to clinical and radiographic resolution of the syndrome. Using this simple procedure allows the surgeon to take advantage of normal temporal horn anatomy and landmarks, avoiding the scarred and distorted region of ventricular obstruction. PMID- 11059676 TI - Unusual intradiploic hematoma. AB - This 58-year-old man presented with a left frontal bone lesion that had been growing over a 2-year period. The lesion increased in size, resulting in proptosis that affected the patient's visual acuity and eyeball movement. On computerized tomography and x-ray studies, a huge lesion located between the widened frontal diploic bone and involving the orbital roof and paranasal sinuses was noted. The entire lesion was radically resected. The authors unexpectedly found that an intradiploic organized hematoma had caused the swelling. PMID- 11059677 TI - Solitary sarcoid granulomatosis mimicking meningioma. PMID- 11059678 TI - Solitary tuberculoid hansen lesion of the ulnar nerve. PMID- 11059679 TI - Extradural posterior inferior cerebellar artery aneurysm. PMID- 11059680 TI - Rupture of an aneurysm during three-dimensional computerized tomography angiography. PMID- 11059681 TI - 3131C--World War II neurosurgery. AB - Preparation for surgical care of the wounded in a two-theater war was extensive and skillfully organized by Michael DeBakey, one of the prime advisors to the Surgeon General of the Army, and by his colleague, Eli Ginzberg, Ph.D. Some of the ways in which this organization was carried out are described. Although the number of neurosurgeons who can recall any involvement of neurosurgery in World War II is diminishing, there remain a significant number who do remember such involvement, many of whom have helped to provide information for this article. PMID- 11059682 TI - Radiosurgery and Cushing's disease. PMID- 11059683 TI - Multiple pituitary adenomas. PMID- 11059684 TI - Pneumoangiography. PMID- 11059685 TI - Long-term follow-up of retroviral vector-administered interferon-gamma (IFN gamma) gene in metastatic melanoma. PMID- 11059686 TI - Lymphoid abnormalities in CD40 ligand transgenic mice suggest the need for tight regulation in gene therapy approaches to hyper immunoglobulin M (IgM) syndrome. AB - Mutations in the CD40 ligand (CD40L) are responsible for human hyper immunoglobulin M (IgM) syndrome. The absence of the interaction between CD40L, expressed by T lymphocytes, and the CD40 receptor present on the surface of B cells is responsible for the inability of B cells to carry out the isotype switch from IgM to the other Ig classes. This leads to a fatal immunodeficiency for which no cure exists. For these reasons, the CD40L gene is a good candidate for gene therapy studies. To investigate the possible effects of the expression of this tightly regulated gene in vivo, we produced transgenic mice in which CD40L expression was deregulated. Widespread ectopic expression appears to be lethal. Overexpression in mature T cells is compatible with life, but in one-third of the cases, mice developed atypical lymphoid proliferations which, occasionally, progressed into frank lymphomas. Even though gene therapy is one of the most promising approaches to cure human hyper IgM syndrome, these results suggest that when we modify very tightly regulated genes such as cytokines or other growth factors, particular care has to be taken to avoid excessive stimulation of the target cells. PMID- 11059687 TI - Adenovirus-mediated antisense ATM gene transfer sensitizes prostate cancer cells to radiation. AB - Treatment failure after radiation therapy of prostate cancer (PC) could be a significant problem. Our objective is to design genetic radiosensitizing strategies for the treatment of PC. Cells from individuals with the genetic disorder ataxia telangiectasia (AT) are hypersensitive to ionizing radiation. Therefore, we examined whether attenuation of the AT gene product, AT mutated (ATM), in PC cells could result in an increased intrinsic radiosensitivity. A p53 mutant PC cell line, PC-3 was infected with adenoviral vectors, expressing antisense ATM RNA to various domains of the ATM gene. Immunoblot analyses of cellular extracts from antisense ATM-transfected PC-3 cells showed attenuated expression of the ATM protein within 2 days of viral infection. Compared with cells infected with an adeno-beta-galactosidase vector, antisense ATM-transfected PC-3 cells showed aberrant control of S-phase cell-cycle checkpoints after exposure to ionizing radiation. Under these conditions, the intrinsic radiosensitivity of the PC-3 cells was enhanced. Consequently antisense ATM gene therapy could serve as a paradigm for strategies that target the cellular survival mechanisms of an irradiated tumor cell and may provide therapeutic benefit to patients undergoing radiation therapy for PC. PMID- 11059688 TI - IkappaBalpha gene therapy in tumor necrosis factor-alpha- and chemotherapy mediated apoptosis of hepatocellular carcinomas. AB - The transcription factor nuclear factor kappaB (NFkappaB) is an essential antagonist of apoptosis during liver regeneration and embryonal development of hepatocytes. Several reports have indicated that NFkappaB may also inhibit the programmed cell death induced by cytokines, ionizing radiation, or cytotoxic drugs in some cancer cell lines. Because hepatocellular carcinomas (HCCs) are one of the most resistant tumors to systemic chemotherapy, we investigated the activation of NFkappaB and the consequence of its inhibition by an IkappaBalpha super repressor during tumor necrosis factor alpha (TNFalpha)- and chemotherapy induced apoptosis in HCC cell lines. We demonstrate that both TNFalpha and adriamycin activate NFkappaB in hepatoma cells. Activation of NFkappaB could be blocked through an adenoviral vector expressing the IkappaBalpha super repressor, regardless of the activating agent. Inhibition of NFkappaB enhanced the apoptosis induced by TNFalpha, whereas IkappaBalpha had an anti-apoptotic effect on chemotherapy-induced programmed cell death. A strong inhibition of chemotherapy- and TNFalpha-induced apoptosis by dominant-negative Fas-associated death domain indicated an essential contribution of death receptor-mediated apoptosis. To elucidate the different role of NFkappaB in chemotherapy-induced apoptosis, we investigated the expression of Fas (CD95) and Fas ligand (CD95 ligand), which have been described as important mediators of chemotherapy-induced cell death and as target genes of NFkappaB. However, our investigations demonstrated that in hepatoma cells, the chemotherapy-induced up-regulation of Fas (CD95) and Fas ligand (CD95 ligand) is not transcriptionally mediated through NFkappaB. Thus, other molecular mechanisms must account for the anti-apoptotic effect of IkappaBalpha in adriamycin-induced death of hepatoma cells. In summary, our investigations indicate that the activation of NFkappaB in response to cytotoxic drugs, in contrast to TNFalpha, exerts a pro-apoptotic stimulus rather than an anti-apoptotic function, which has implications for therapy of HCCs. PMID- 11059689 TI - Nitric oxide-mediated tumor cell killing of cisplatin-based interferon-gamma gene therapy in murine ovarian carcinoma. AB - We have determined the role of nitric oxide (NO)-mediated tumor cell killing in the treatment of an animal model of murine ovarian carcinoma grown in the peritoneum with a combination of cisplatin and cationic liposomes containing an expression vector for interferon-gamma (IFN-gamma). The approach was to determine whether the therapy was effective in mice homozygous for a disrupted inducible NO synthase (iNOS) allele; these mice were unable to produce NO in response to IFN gamma. iNOS (-/-) mice treated with both cisplatin and liposomal IFN-gamma gene did not produce a significant amount of NO in ascites (12.1+/-4.5 microM), although they expressed a high level of IFN-gamma (9002+/-723 U/mL of ascitic fluid). As a result, mice died of tumors within 11-62 days. However, wild-type mice treated with both cisplatin and liposomal IFN-gamma gene produced a significant amount of NO in ascites (113.7+/-17.9 microM) with a high level of IFN-gamma gene expression (9350+/-1254 U/mL of ascitic fluid) and were free of tumors for at least 80 days. This result confirmed that NO was a direct mediator of IFN-gamma cytotoxicity. PMID- 11059690 TI - Enhancement of adenoviral transduction with polycationic liposomes in vivo. AB - Although the high transfection efficiency with adenovirus in vitro is well documented, it is still not clear whether adenoviral vectors are effective in vivo in solid tumor models. In our preliminary experiment, transduction of tumor tissue was limited to just around the injection site after intratumoral injection of the adenoviral vector. To improve the transduction efficiency in vivo, we tried a combination of adenoviral vector and liposome in our animal model. Adenovirus carrying human placental alkaline phosphatase (AdALP) and Lipofectamine or 1,3-di-oleoyloxy-2-(6-carboxyspermyl)-propylamide were used as a marker gene and the cationic liposome, respectively. A >15-fold increase in the transfection efficiency was observed in CT26 tumor cell lines with the combination of AdALP adenovirus carrying murine granulocyte-macrophage colony stimulating factor (AdmGM-CSF), and liposome compared with adenovirus alone, showing the feasibility of the combination treatment. In the animal model, with the combination of liposome and AdALP, deeper and wider distribution of the marker gene in the tumor mass was shown. We conclude that the limitations of direct application of adenoviral vectors in a solid tumor model could be overcome by the use of cationic liposomes. This approach will facilitate the more effective delivery of adenoviral vectors in a clinical trial setting. PMID- 11059691 TI - In vivo tumor delivery of the green fluorescent protein gene to report future occurrence of metastasis. AB - The green fluorescent protein (GFP) gene was administered to intraperitoneally (i.p.) growing human stomach cancer in nude mice to visualize future regional and distant metastases. GFP retroviral supernatants were injected i.p. from day 4 to day 10 after i.p. implantation of the cancer cells. Tumor and metastasis fluorescence was visualized every other week with the use of fluorescence optics via a laparotomy on the tumor-bearing animals. At 2 weeks after retroviral GFP delivery, GFP-expressing tumor cells were observed in gonadal fat, greater omentum, and intestine, indicating that these primary i.p. growing tumors were efficiently transduced by the GFP gene and could be visualized by its expression. At the second and third laparotomies, GFP-expressing tumor cells were observed to have spread to lymph nodes in the mesentery and other regional sites. At the fourth laparotomy, widespread tumor growth was visualized by GFP expression, inducing liver metastasis. No normal tissues were found to be transduced by the GFP retrovirus. Thus, reporter gene transduction of the primary tumor enabled detection of its subsequent metastasis. This gene therapy model could be applied to primary tumors before resection or other treatment to have a fluorescent early detection system for metastasis and recurrence. PMID- 11059692 TI - A novel strategy for cancer therapy by mutated mammalian degenerin gene transfer. AB - Mammalian degenerin (MDEG) is a member of the amiloride-sensitive sodium ion channel family, and its site-directed active mutant (MDEG-G430F) induces massive Na+ influx into cells, leading to cell ballooning and cell bursting. We attempted a novel therapeutic approach for gastric cancers by transferring MDEG-G430F into cancer cells using tumor-specific promoters. In carcinoembryonic antigen (CEA) producing gastric cancer cells, the level of cell death observed when MDEG-G430F was used with a CEA promoter was similar to that observed when using a potent nonspecific promoter such as the cytomegalovirus promoter. In an in vivo study, fusogenic liposome complexes containing MDEG-G430F driven by the CEA promoter were injected intraperitoneally into CEA-producing gastric cancer cells in a mouse peritoneal dissemination model. Although all 15 of the control mice were dead by 50 days postinoculation, 13 of the 15 mice treated with MDEG-G430F survived. These results indicate that transferring MDEG-G430F into cancer tissues using tumor-specific promoters can achieve striking and selective cancer cell death irrespective of the transcriptional efficiency of the promoters used in vivo, and suggest that this approach is a promising new strategy for cancer gene therapy. PMID- 11059693 TI - In suicide gene therapy, the site of subcellular localization of the activating enzyme is more important than the rate at which it activates prodrug. AB - The bacterial enzyme carboxypeptidase G2 (CPG2) can be expressed both intracellularly (CPG2*) or tethered to the outer surface (stCPG2(Q)3) of mammalian cells, where it is able to activate mustard prodrugs for use in suicide gene therapy protocols. Here we compare the properties of CPG2 expressed in these two locations. CPG2 is active as a dimer, and one of the mutations required to block glycosylation of stCPG2(Q)3 destabilizes the dimers. Some of the mutations to this site partially correct the dimerization defect and recover a proportion of the activity. Surface tethering also recovers some enzyme activity, but through an unknown mechanism. The efficacy of CPG2 in these two locations is compared with the tumor cell lines A2780, SK-OV-3, and WiDr, which are sensitized to the prodrug 4-([2-chloroethyl][2-mesyloxyethyl]amino)benzoyl-L-glutamic acid (CMDA) by both CPG2* and stCPG2(Q)3 expression in suicide gene therapy protocols in vitro. We find that stCPG2(Q)3 is a more efficient mediator of CMDA-dependent cell killing than CPG2*. Lower levels of stCPG2(Q)3 activity are required to give cell killing that can only be achieved by higher levels of CPG2*. In bystander effect assays, low levels of stCPG2(Q)3 are required for efficient killing, whereas relatively high levels of CPG2* activity are required. Also, shorter exposures to prodrug are required for cell killing when stCPG2(Q)3 is expressed compared with when CPG2* is expressed. These data demonstrate that the location of the enzyme in the cell is more important than the enzyme activity as the determinant in mediating cytotoxicity. PMID- 11059694 TI - Induction of protective immunity against syngeneic rat cancer cells by expression of the cytosine deaminase suicide gene. AB - The use of the cytosine deaminase (CD)/5-fluorocytosine suicide system as a cancer gene therapy approach enables selective killing of CD-modified cells as well as the ablation of non-modified tumor cells due to a bystander effect that has been suggested to involve the immune system in vivo. Using a stable CD transfectant of the tumorigenic rat adenocarcinoma cell line AS (AS/CD), an antitumoral response against the CD expressing cell line as well as the parental cell line could be induced by stepwise vaccinations in syngeneic animals. AS/CD tumor regression occurred independently of 5-fluorocytosine treatment and was sufficient to protect 37% of the animals against subsequent challenge with tumorigenic doses of the parental AS cell line. Immune rats contained lymphocytes able to specifically lyse CD modified as well as unmodified AS tumor cells in vitro, most likely contributing to the in vivo antitumoral reaction. Thus, the CD suicide system seems to be suitable not only for a local tumor gene therapy but also for the application as therapy of metastatic tumors and minimal residual disease. PMID- 11059695 TI - Vaccination with B16 melanoma cells expressing a secreted form of interleukin 1beta induces tumor growth inhibition and an enhanced immunity against the wild type B16 tumor. AB - We have demonstrated previously that gene transfer of the mature human interleukin-1beta (IL-1beta) gene, fused to a signal sequence (ss), into mouse B16 melanoma cells results in an inhibition of their growth in vivo compared with control B16 cells. We here extend these results to show that intraperitoneal vaccinations with irradiated IL-1beta-secreting cells result in protection against subsequent subcutaneous challenge with wild-type (wt) B16 tumor cells in syngeneic C57BL/6 mice. This protection appears to be long-lasting, because rechallenge of cured mice 4 months after the first challenge also demonstrated resistance. In addition, we demonstrate that mice with established wt tumors subjected to therapeutic vaccinations with irradiated B16/ssIL-1beta cells starting 3 days after challenge isografting have a significantly inhibited tumor growth and 25-40% survival at the challenge doses given. In vitro coculture of spleen cells from B16/ssIL-1beta vaccinated animals and wt B16 cells induced an enhanced proliferative response, which correlated with elevated production of IL 2 and interferon-gamma. A significantly enhanced cytolytic activity against B16 wt target tumor cells was observed when spleen cells from B16/ssIL-1beta vaccinated mice were used as effector cells compared with spleen cells from control vaccinated mice. In vitro depletion experiments using anti-asialo GM1 revealed a prominent role for natural killer cells as effector cells. The data suggest that local IL-1beta secretion during the vaccination phase can provoke or augment protective immune responses to B16 melanoma cells, which are otherwise not recorded in mice bearing B16 tumors. PMID- 11059696 TI - Identification of neonatal hearing impairment: a cornerstone for newborn hearing screening. PMID- 11059697 TI - Identification of neonatal hearing impairment: a multicenter investigation. AB - OBJECTIVES: This article describes the design of a multicenter study sponsored by the National Institutes of Health. The purpose of this study was to determine the accuracy of three measures of peripheral auditory system status (transient evoked otoacoustic emissions, distortion product otoacoustic emissions, and auditory brain stem responses) applied in the perinatal period for predicting behavioral hearing status at 8 to 12 mo corrected age. The influences of the infant's medical status, the test environment, and test and response parameters on test performance were examined. DESIGN: Seven institutions participated in this study. There were 7179 infants evaluated in the perinatal period. All graduates of the neonatal intensive care unit (4478) and well babies with one or more risk factor for hearing loss (353) were targeted for follow-up testing using visual reinforcement audiometry (VRA) at 8 to 12 mo corrected age. Well babies without any risk indicators (N = 2348) were not targeted for follow-up VRA testing. However, 80 of these well babies did not pass the screening protocol and thus were targeted for follow-up VRA testing as well. Perinatal test performance was evaluated using the VRA data as the "gold standard." RESULTS: The results of this study are described in a series of 11 articles following this introductory article. CONCLUSIONS: The evaluation of newborn hearing tests required a longitudinal study in which newborn test results were compared with a gold standard based on behavioral audiometric assessment. Such an evaluation was possible because all newborns, passes as well as refers, were followed up long enough to permit reliable behavioral measurements. In addition, prenatal, perinatal, and maternal history information, test environment, and test parameter information were collected to provide data that led to a complete description of factors affecting test outcomes. All of these data were obtained in a sample of sufficient ethnic, medical, and geographic diversity in efforts to increase the generalizability of the results. Finally, the data were combined in a relational data base to examine the factors that influence test performance. Specific information related to these issues is presented in the articles that follow. PMID- 11059698 TI - Identification of neonatal hearing impairment: experimental protocol and database management. AB - OBJECTIVE: The purposes of this article are to describe the overall protocol for the Identification of Neonatal Hearing Impairment (INHI) project and to describe the management of the data collected as part of this project. A well-defined protocol and database management techniques were needed to ensure that data were 1) collected accurately and in the same way across sites; 2) maintained in a database that could be used to provide feedback to individual sites regarding enrollment and the extent to which the protocol was complete on individual subjects; and 3) available to answer project questions. This article describes techniques that were used to meet these needs. DESIGN: This study was a prospective, randomized study that was designed to evaluate auditory brain stem responses, transient evoked otoacoustic emissions, and distortion product otoacoustic emissions as hearing-screening tools, and to relate neonatal test findings to hearing status, defined by visual reinforcement audiometry at 8 to 12 mo of age. Measures of middle-ear function also were obtained at some sites as part of the neonatal test battery. In addition, other clinical and demographic data were gathered to determine the extent to which factors, other than auditory status, influenced test behavior. Three groups were evaluated: neonatal intensive care unit (NICU) infants (those who spent 3 or more days in a NICU), well babies with risk factors for hearing loss, and well babies without risk factors. Six centers participated in the trial. The testers for the project included audiologists, technicians, audiology graduate students, and medical research staff. The same computerized neonatal test program was applied at each center. This program generated the neonatal test database automatically. Clinical and demographic data were collected by means of concise data collection forms and were entered into a database at each site. After the neonatal test, subjects from the NICU and at-risk well babies were evaluated with visual reinforcement audiometry starting at 8 to 12 mo of age. All data were electronically transmitted to the core site where they were merged into one overall database. This database was exercised to provide feedback and to identify discrepancies throughout the course of the study. In its final form, it served as the database on which all analyses were performed. RESULTS AND CONCLUSION: The protocol was a departure from typical hearing screening procedures in terms of 1) its regimented application of three screening measures; 2) the detailed information that was obtained regarding subject clinical and demographic factors; and 3) its application of the same procedures across six centers having diverse geographic location and subject demographics. A learning curve for successfully executing the study protocols was observed. Throughout the study, monthly reports were generated to monitor subject enrollment, check for data completeness, and to perform data integrity checks. In combination with monthly data reports and checks that occurred throughout the progression of the study, miscellaneous data audits were performed to check accuracy of neonatal testing programs and to cross check information entered in the clinical and demographic database. The data management techniques used in this project helped to ensure the quality of the data collection process and also allowed for detailed analyses once data were collected. This was particularly important because it enabled us to evaluate not only the performance of individual measures as screening tools, but also permitted an evaluation of the influence of other variables on screening test results. PMID- 11059699 TI - Identification of neonatal hearing impairment: characteristics of infants in the neonatal intensive care unit and well-baby nursery. AB - OBJECTIVE: The objective of this study was to describe the demographic data, medical status, and incidence of risk factors for hearing impairment in the neonatal intensive care unit (NICU) and well-baby populations in a multicenter prospective study designed to assess neonatal hearing impairment and to evaluate factors that might affect neonatal hearing test performance. DESIGN: This was a prospective multicenter study funded by the National Institutes of Health National Institute on Deafness and Other Communication Disorders to evaluate the effectiveness of auditory brain stem response, transient evoked otoacoustic emissions, and distortion product otoacoustic emissions for newborn hearing screening. Research staff at each site obtained informed consent and detailed demographic and medical data, including information on established risk factors for hearing loss on 4478 high-risk infants cared for in the NICU, 2348 infants from the well-baby nurseries with no risk factor, and 353 infants from the well baby nurseries with risk factors. For follow-up purposes the sample was divided further to include a subgroup called selects. Selects were either infants from the well-baby nursery who had an established risk factor for hearing impairment (N = 353) or did not pass the neonatal hearing screen protocol (N = 80). In this study, we focus on the distribution of infants by nursery and risk factors only. Particular effort was made to enroll infants with risk factors for hearing loss in both the NICU and well-baby nurseries. Descriptive analyses are used to describe characteristics of this sample. RESULTS: All 10 of the risk factors established by the Joint Committee on Infant Hearing in 1994 were identified in the NICU population. The four most common were ototoxic medications (44.4%), very low birth weight (17.8%), assisted ventilation > 5 days (16.4%), and low Apgar scores at 1 or 5 min (13.9%). In contrast, only six risk factors were present in the well-baby nurseries: family history (6.6%), craniofacial abnormalities (3.4%), low Apgar scores (2.8%), syndromes (0.5%), ototoxic medications (0.2%), and congenital infection (0.1%). CONCLUSION: These descriptive risk factor data reflect both the newborn populations at the study sites and the bias for enrolling infants at risk for hearing loss. The high-risk NICU sample reflects the characteristics typically found in graduates of the NICU. The data summarized in this study will be used to assess the relationships between risk factor and hearing test outcome. PMID- 11059700 TI - Identification of neonatal hearing impairment: auditory brain stem responses in the perinatal period. AB - OBJECTIVES: 1) To describe the auditory brain stem response (ABR) measurement system and optimized methods used for study of newborn hearing screening. 2) To determine how recording and infant factors related to the screening, using well defined, specific ABR outcome measures. DESIGN: Seven thousand one hundred seventy-nine infants, 4478 from the neonatal intensive care unit (NICU) and the remaining from the well-baby nursery, were evaluated with an automated ABR protocol in each ear. Two channel recordings were obtained (vertex to mastoid or channel A and vertex to nape of neck or channel B) in response to click stimuli of 30 and 69 dB nHL in all infants as well as 50 dB nHL in infants who did not meet criteria for response at 30 dB. Criteria for response included F(SP) > or =3.1 and a tester-judgment of response. Criteria could be met in the first or repeat test with a maximum of 6144 accepted sweeps per test. RESULTS: More than 99% of infants could complete the ABR protocol. More than 90% of NICU and well baby nursery infants "passed" given the strict criteria for response, whereas 86% of those with high risk factors met criterion for ABR response detection. The number of infants who did not meet ABR response criteria in one or both ears was systematically related to stimulus level with the largest group not meeting criteria at 30 dB followed by 50 and 69 dB nHL. Meeting criteria on the ABR was positively correlated with the amplitude of wave V, with low noise and low electrode impedance. Factors that predicted how many sweeps would be needed to reach criterion F(SP) included noise level of the test site, state of the baby (for example, quiet sleep versus crying), recording noise, electrode impedance and response latency. Channel A (vertex to mastoid) reached criterion more often than channel B (vertex to nape of neck) due to higher noise in channel B. Average total test time for 30 dB nHL screening in both ears was under 8 minutes. Well babies with risk factors took slightly longer to evaluate than other groups with this automated ABR procedure and have higher noise levels. CONCLUSIONS: ABR implemented with an automated detection algorithm using a 30 dB nHL click stimulus is reliable technique for rapid assessment of auditory status in newborns. Factors other than hearing loss that influenced the test result include infant state, electrode location and impedance, testing site, and infant risk status. Even so, ABRs were reliably recorded in the vast majority of babies under circumstances in which most babies are found in the perinatal period. PMID- 11059701 TI - Identification of neonatal hearing impairment: distortion product otoacoustic emissions during the perinatal period. AB - OBJECTIVES: 1) To describe distortion product otoacoustic emission (DPOAE) levels, noise levels and signal to noise ratios (SNRs) for a wide range of frequencies and two stimulus levels in neonates and infants. 2) To describe the relations between these DPOAE measurements and age, test environment, baby state, and test time. DESIGN: DPOAEs were measured in 2348 well babies without risk indicators, 353 well babies with at least one risk indicator, and 4478 graduates of neonatal intensive care units (NICUs). DPOAE and noise levels were measured at f2 frequencies of 1.0, 1.5, 2.0, 3.0, and 4.0 kHz, and for primary levels (L1/L2) of 65/50 dB SPL and 75/75 dB SPL. Measurement-based stopping rules were used such that a test did not terminate unless the response was at least 3 dB above the mean noise floor + 2 SDs (SNR) for at least four of five test frequencies. The test would terminate, however, if these criteria were not met after 360 sec. Baby state, test environment, and other test factors were captured at the time of each test. RESULTS: DPOAE levels, noise levels and SNRs were similar for well babies without risk indicators, well babies with risk indicators, and NICU graduates. There was a tendency for larger responses at f2 frequencies of 1.5 and 2.0 Hz, compared with 3.0 and 4.0 kHz; however, the noise levels systematically decreased as frequency increased, resulting in the most favorable SNRs at 3.0 and 4.0 kHz. Response levels were least and noise levels highest for an f2 frequency of 1.0 kHz. In addition, test time to achieve automatic stopping criteria was greatest for 1.0 kHz. With the exception of "active/alert" and "crying" babies, baby state had little influence on DPOAE measurements. Additionally, test environment had little impact on these measurements, at least for the environments in which babies were tested in this study. However, the lowest SNRs were observed for infants who were tested in functioning isolettes. Finally, there were some subtle age affects on DPOAE levels, with the infants born most prematurely producing the smallest responses, regardless of age at the time of test. CONCLUSIONS: DPOAE measurements in neonates and infants result in robust responses in the vast majority of ears for f2 frequencies of at least 2.0, 3.0 and 4.0 kHz. SNRs decrease as frequency decreases, making the measurements less reliable at 1.0 kHz. When considered along with test time, there may be little justification for including an f2 frequency at 1.0 kHz in newborn screening programs. It would appear that DPOAEs result in reliable measurements when tests are conducted in the environments in which babies typically are found. Finally, these data suggest that babies can be tested in those states of arousal that are most commonly encountered in the perinatal period. PMID- 11059702 TI - Identification of neonatal hearing impairment: transient evoked otoacoustic emissions during the perinatal period. AB - OBJECTIVES: 1) To describe transient evoked otoacoustic emission (TEOAE) levels, noise levels and signal to noise ratios (SNRs) for a range of frequency bands in three groups of neonates who were tested as a part of the Identification of Neonatal Hearing Impairment multi-center consortium project. 2) To describe the relations between these TEOAE measurements and age, test environment, baby state, and test time. DESIGN: TEOAEs were measured in 4478 graduates of neonatal intensive care units (NICUs), 353 well babies with at least one risk indicator, and 2348 well babies without risk factors. TEOAE and noise levels were measured for frequency bands centered at 1.0, 1.5, 2.0, 3.0, and 4.0 kHz for a click stimulus level of 80 dB SPL. For those ears not meeting "passing" stopping criteria at 80 dB pSPL, a level of 86 dB pSPL was included. Measurement-based stopping rules were used such that a test did not terminate unless the response revealed a criterion SNR in four out of five frequency bands or no response occurred after a preset number of averages. Baby state, test environment, and other test factors were captured at the time of test. RESULTS: TEOAE levels, noise levels and SNRs were similar for NICU graduates, well babies with risk factors and well babies without risk factors. There were no consistent differences in response quality as a function of test environment, i.e., private room, unit, open crib, nonworking isolette, or working isolette. Noise level varied little across risk group, test environment, or infant state other than crying, suggesting that the primary source of noise in TEOAE measurements is infant noise. The most significant effect on response quality was center frequency. Responses were difficult to measure in the half-octave band centered at 1.0 kHz, compared with higher frequencies. Reliable responses were measured routinely at frequencies of 1.5 kHz and higher. CONCLUSIONS: TEOAEs are easily measured in both NICU graduates and well babies with and without risk factors for hearing loss in a wide variety of test environments. Given the difficulties encountered in making reliable measurements for a frequency band centered at 1.0 kHz, its inclusion in a screening program may not be justified. PMID- 11059703 TI - Identification of neonatal hearing impairment: ear-canal measurements of acoustic admittance and reflectance in neonates. AB - OBJECTIVES: 1) To describe broad bandwidth measurements of acoustic admittance (Y) and energy reflectance (R) in the ear canals of neonates. 2) To describe a means for evaluating when a YR response is valid. 3) To describe the relations between these YR measurements and age, gender, left/right ear, and selected risk factors. DESIGN: YR responses were obtained at four test sites in well babies without risk indicators, well babies with at least one risk indicator, and graduates of neonatal intensive care units. YR responses were measured using a chirp stimulus at moderate levels over a frequency range from 250 to 8000 Hz. The system was calibrated based on measurements in a set of cylindrical tubes. The probe assembly was inserted in the ear canal of the neonate, and customized software was used for data acquisition. RESULTS: YR responses were measured in over 4000 ears, and half of the responses were used in exploratory data analyses. The particular YR variables chosen for analysis were energy reflectance, equivalent volume and acoustic conductance. Based on the view that unduly large negative equivalent volumes at low frequencies were physically impossible, it was concluded that approximately 13% of the YR responses showed evidence of improper probe seal in the ear canal. To test how these outliers influenced the overall pattern of YR responses, analyses were conducted both on the full data set (N = 2081) and the data set excluding outliers (N = 1825). The YR responses averaged over frequency varied with conceptional age (conception to date of test), gender, left/right ear, and selected risk factors; in all cases, significant effects were observed more frequently in the data set excluding outliers. After excluding outliers and controlling for conceptional age effects, the dichotomous risk factors accounting for the greatest variance in the YR responses were, in rank order, cleft lip and palate, aminoglycoside therapy, low birth weight, history of ventilation, and low APGAR scores. In separate analyses, YR responses varied in the first few days after birth. An analysis showed that the use of a YR test criterion to assess the quality of probe seal may help control the false-positive rate in evoked otoacoustic emission testing. CONCLUSIONS: This is the first report of wideband YR responses in neonates. Data were acquired in a few seconds, but the responses are highly sensitive to whether the probe is fully sealed in the ear canal. A real-time acoustic test of probe fit is proposed to better address the probe seal problem. The YR responses provide information on middle ear status that varies over the neonatal age range and that is sensitive to the presence or absence of risk factors, ear, and gender differences. Thus, a YR test may have potential for use in neonatal screening tests for hearing loss. PMID- 11059704 TI - Identification of neonatal hearing impairment: recruitment and follow-up. AB - OBJECTIVE: The purpose of this study is to describe the recruitment and retention strategies as well as the sample demographics for families with infants completing the neonatal examination and returning for follow-up. These data are compared to those infants inactivated from the study. DESIGN: This study was a prospective, randomized clinical study. All infants who were confined to the neonatal intensive care unit (NICU) and well babies with at least one risk indicator were targeted for behavioral audiometric follow-up testing. In addition, infants without risk factors from the well-baby nursery, but who failed a newborn test, were also followed. Several variables were evaluated to determine those factors, if any, that might predict which families returned for follow-up testing. RESULTS: Recruitment was achieved as per study design with 4911 high risk infants and 2348 well-baby nursery infants (without risk indicators for hearing) enrolled. Of the 4911 high-risk infants enrolled, 64% were successfully recruited into the follow-up portion of the study. This was less than the projected rate of 80%. Factors predicting noncompliance with the study protocol for follow-up were predominantly sociodemographic and included nonwhite race, no insurance, substance abuse, young maternal age, more than two children at home, and late onset of prenatal care. CONCLUSIONS: Factors related to low socioeconomic status and increased social risk were the strongest predictors of poor study protocol compliance. Despite retention challenges, 64% of the targeted, high-risk infants subsequently returned for the 8-to 12-mo behavioral hearing assessment protocols for validation purposes. PMID- 11059705 TI - Identification of neonatal hearing impairment: hearing status at 8 to 12 months corrected age using a visual reinforcement audiometry protocol. AB - OBJECTIVES: 1) To describe the hearing status of the at-risk infants in the National Institutes of Health-Identification of Neonatal Hearing Impairment study sample at 8 to 12 mo corrected age (chronologic age adjusted for prematurity). 2) To describe the visual reinforcement audiometry (VRA) protocol that was used to obtain monaural behavioral data for the sample. DESIGN: All neonatal intensive care unit infants and well babies with risk factors (including well babies who failed neonatal tests) were targeted for follow-up behavioral evaluation once they had reached 8 mo corrected age. Three thousand one hundred and thirty-four (64.4%) of the 4868 surviving infants returned for at least one behavioral hearing evaluation, which employed a well-defined VRA protocol. VRA thresholds or minimum response levels (MRLs) were determined for speech and pure tones of 1.0, 2.0, and 4.0 kHz for each ear using insert earphones. RESULTS: More than 95% of the infants were reliably tested with the VRA protocol; 90% provided complete tests (four MRLs for both ears). Ninety-four percent of the at-risk infants were found to have normal hearing sensitivity (MRLs of 20 dB HL) at 1.0, 2.0, and 4.0 kHz in both ears. Of the infants, 2.2% had bilateral hearing impairment, and 3.4% had impairment in one ear only. More than 80% of the impaired ears had losses of mild-to-moderate degree. CONCLUSIONS: This may be the largest study to attempt to follow all at-risk infants with behavioral audiometric testing, regardless of screening outcome, in an effort to validate the results of auditory brain stem response, distortion product otoacoustic emission, and transient evoked otoacoustic emission testing in the newborn period. It is one of only a few studies to report hearing status of infants at 1 yr of age, using VRA on a clinical population. Successful testing of more than 95% of the infants who returned for the VRA follow-up documents the feasibility of obtaining monaural behavioral data in this population. PMID- 11059706 TI - Identification of neonatal hearing impairment: infants with hearing loss. AB - OBJECTIVE: This article describes the audiologic findings and medical status of infants who were found to have hearing loss, detected as part of the Identification of Neonatal Hearing Impairment (INHI) project. In addition, the neonatal and maternal health variables for the group of infants who could not be tested with visual reinforcement audiometry (VRA) due to developmental and visual disability are presented. DESIGN: The overall goal of the INHI project was to evaluate the test performance of auditory brain stem response and evoked otoacoustic emission (OAE) tests given in the newborn period. These tools were evaluated on the basis of the infants' hearing when tested behaviorally with VRA at 8 to 12 mo corrected age. The neonatal test results, VRA results, medical history information and a record of intercurrent events occurring between the neonatal period and the time of VRA were collated and reviewed. The purpose of this article is to review the characteristics of those infants who were found to have hearing loss. RESULTS: Of 2995 infants who had VRA tests judged to be of good or fair reliability, 168 had a finding of hearing loss for at least one ear, an incidence of 5.6%. Sixty-six infants had bilateral losses, an incidence of 2%, and 22 infants had bilateral hearing losses in the moderate to profound range, an incidence 0.7%. The prevalence of middle ear problems was greater than 50% among these infants with hearing loss. From the larger group of 168 infants with hearing loss, a group of 56 infants (86 ears) was chosen as those with a low probability that the hearing loss was due to transient middle ear pathology and was more likely hearing loss of a permanent nature. These were the infants used for the analyses of neonatal test performance (Norton et al., 2000). In this selected group there were 30 infants with bilateral impairment of at least mild degree, which is an incidence of 1%. There were approximately equal numbers of ears in the mild, moderate, severe and profound range of hearing loss. Risk factors associated with hearing loss were reviewed for the total sample of infants tested with VRA and for those infants with hearing loss. A history of treatment with aminoglycosides was the risk factor most often reported in the entire sample; however, there was no difference in prevalence of this risk factor for the normal-hearing and hearing-impaired groups. The risk factor associated with the highest incidence of hearing loss was stigmata of syndromes associated with sensorineural hearing loss and other neurosensory disorders. Sixty-seven infants who returned for follow-up could not be tested with VRA due to severe developmental delay or visual disability. Many of these infants had medical histories indicating the sequelae of extreme prematurity and/or very low birthweight. CONCLUSIONS: Most of the hearing losses found in this study were mild and, based on clinical history and tympanometry tests, many of the mild and some of the moderate impairments may have been acquired in early infancy due to middle ear effusion. In the group of infants used for determination of neonatal test performance there were approximately equal numbers of mild, moderate, severe and profound losses. Only a small percentage of infants with a conventional risk indicator for hearing loss actually had a hearing loss, and there were a significant number of infants with hearing loss who did not have a risk indicator. These findings support the need for an early identification program based on universal neonatal hearing screening rather than by targeted testing of those with risk indicators. PMID- 11059707 TI - Identification of neonatal hearing impairment: evaluation of transient evoked otoacoustic emission, distortion product otoacoustic emission, and auditory brain stem response test performance. AB - OBJECTIVES: The purpose of this study was to compare the performance of transient evoked otoacoustic emissions (TEOAEs), distortion product otoacoustic emissions (DPOAEs), and auditory brain stem responses (ABRs) as tools for identification of neonatal hearing impairment. DESIGN: A total of 4911 infants including 4478 graduates of neonatal intensive care units, 353 well babies with one or more risk factors for hearing loss (Joint Committee on Infant Hearing, 1994) and 80 well babies without risk factor who did not pass one or more neonatal test were targeted as the potential subject pool on which test performance would be assessed. During the neonatal period, they were evaluated using TEOAEs in response to an 80 dB pSPL click, DPOAE responses to two stimulus conditions (L1 = L2 = 75 dB SPL and L1 = 65 dB SPL L2 = 50 dB SPL), and ABR elicited by a 30 dB nHL click. In an effort to describe test performance, these "at-risk" infants were asked to return for behavioral audiologic assessments, using visual reinforcement audiometry (VRA) at 8 to 12 mo corrected age, regardless of neonatal test results. Sixty-four percent of these subjects returned and reliable VRA data were obtained on 95.6% of these returnees. This approach is in contrast to previous studies in which, by necessity, efforts were made to follow only those infants who "failed" the neonatal screening tests. The accuracy of the neonatal measures in predicting hearing status at 8 to 12 mo corrected age was determined. Only those infants who provided reliable, monaural VRA test results were included in the analysis. Separate analyses were performed without regard to intercurrent events (i.e., events between the neonatal and VRA tests that could cause their results to disagree), and then after accounting for the possible influence of intercurrent events such as otitis media and late-onset or progressive hearing loss. RESULTS: Low refer rates were achieved for the stopping criteria used in the present study, especially when a protocol similar to the one recommended in the National Institutes of Health (1993) Consensus Conference Report was followed. These analyses, however, do not completely describe test performance because they did not compare neonatal screening test results with a gold standard test of hearing. Test performance, as measured by the area under a relative operating characteristic curve, were similar for all three neonatal tests when neonatal test results were compared with VRA data obtained at 8 to 12 mo corrected age. However, ABRs were more successful at determining auditory status at 1 kHz, compared with the otoacoustic emission (OAE) tests. Performance was more similar across all three tests when they were used to identify hearing loss at 2 and 4 kHz. No test performed perfectly. Using either the two- or three frequency pure-tone average (PTA), with a fixed false alarm rate of 20%, hit rates for the neonatal tests, in general, exceeded 80% when hearing impairment was defined as behavioral thresholds > or =30 dB HL. All three tests performed similarly when a two-frequency (2 and 4 kHz) PTA was used as the gold standard; OAE test performance decreased when a three-frequency PTA (adding 1 kHz) was used as the gold standard definition. For both PTA and all three neonatal screening measures, however, hit rate increased as the magnitude of hearing loss increased. CONCLUSIONS: Singly, all three neonatal hearing screening tests resulted in low refer rates, especially if referrals for follow-up were made only for the cases in which stopping criteria were not met in both ears. Following a protocol similar to that recommended in the National Institutes of Health (1993) Consensus Conference report resulted in refer rates that were less than 4%. TEOAEs at 80 dB pSPL, DPOAE at L1 = 65, L2 = 50 dB SPL and ABR at 30 dB nHL measured during the neonatal period, and as implemented in the current study, performed similarly at predicting behavioral hearing status at 8 to 12 PMID- 11059708 TI - Identification of neonatal hearing impairment: summary and recommendations. AB - OBJECTIVES: This article summarizes the results of a multi-center study, "Identification of Neonatal Hearing Impairment," sponsored by the National Institutes of Health. The purpose of this study was to determine the performance characteristics of three measures of peripheral auditory system status, transient evoked otoacoustic emissions (TEOAEs), distortion product otoacoustic emissions (DPOAEs), and auditory brain stem responses (ABR), applied in the neonatal period in predicting hearing status at 8 to 12 mo corrected age. DESIGN: The design and implementation of this study are described in the first two articles in this series. Seven institutions participated in this study; 7179 infants were evaluated. Graduates of the neonatal intensive care unit and well babies with one or more risk factors for hearing loss were targeted for follow-up testing using visual reinforcement audiometry (VRA) at 8 to 12 mo corrected age. Neonatal test performance was evaluated using the VRA data as the "gold standard." RESULTS: The major results of the study are described in the nine articles preceding this summary article. TEOAEs in response to an 80 dB pSPL click, DPOAEs in response to L1 = 65 and L2 = 50 dB SPL and ABR in response to a 30 dB nHL click performed well as predictors of permanent hearing loss of 30 dB or greater at 8 to 12 mo corrected age. All measures were robust with respect to infant state, test environment and infant medical status. No test performed perfectly. CONCLUSIONS: Based on the data from this study, the 1993 National Institutes of Health Consensus Conference-recommended protocol-an OAE test followed by an ABR test for those infants failing the OAE test-would result in low referral rate (96 to 98%). TEOAEs for 80 dB pSPL, ABR for 30 dB nHL and DPOAEs for L1 = 65 dB SPL and L2 = 50 dB SPL perform well in predicting hearing status based on the area under the relative operating characteristic curve. Accuracy for the OAE measurements are best when the speech awareness threshold or the pure-tone average for 2.0 kHz and 4 kHz are used as the gold standard. ABR accuracy varies little as a function of the frequencies included in the gold standard. In addition, 96% of those infants returning for VRA at 8 to 12 mo corrected age were able to provide reliable ear specific behavioral thresholds using insert earphones and a rigorous psychophysical VRA protocol. PMID- 11059709 TI - Endoscopic plastic surgery. AB - This article discusses three of the most popular endoscopic procedures in plastic surgery. Brow lift, transaxillary breast augmentation, and abdominoplasty are all cosmetic procedures with a high demand on inconspicuous scars; however, many investigators are working on reconstructive endoscopically assisted procedures. The treatment of many facial fractures involving the upper third of the facial skeleton usually requires long bicoronal incisions similar to the incisions used in the traditional brow lift. Attempts are under way to use endoscopically assisted minimal-access techniques to reduce and fixate these fractures. Many flaps used in plastic surgery require long scars for harvest, as in the case of the latissimus dorsi muscle flap. A relatively long incision on the back is needed to gain access to the muscle so that it can be elevated from structures superficial and deep to it. Although it is unpopular, investigators have reported harvesting latissimus dorsi muscle flaps through fairly small incisions with the assistance of balloon dissectors and endoscopes. In the field of hand surgery, carpal tunnel release surgery has had more than one method proposed to transect the carpal ligament using endoscopes and special instrumentation. Although some reported series claim excellent results, many hand surgeons are reluctant to use endoscopes because of associated complications and a high recurrence rate of carpal tunnel syndrome. Plastic surgery has special demands that emphasize aesthetics in cosmetic and reconstructive procedures. Although the lack of natural optical cavities has slowed the incorporation of endoscopic surgery in the specialty, surgically created cavities are used to allow for minimal access incisions. The future of plastic surgery will include an ever-increasing number of endoscopically assisted procedures. Cosmetic and reconstructive procedures will benefit from this new technology. PMID- 11059710 TI - Minimal access breast surgery. AB - Minimal access procedures have great potential for providing patients with equal, if not superior, forms of breast cancer diagnosis and treatment. Many of these procedures are in a process of evolution. The reliability of each method probably depends heavily on the training, ability, and experience of the operator. Surgeons should be aware of the advantages and pitfalls of these techniques and exercise caution during the initial phases of their learning experience. PMID- 11059711 TI - Minimally invasive parathyroid surgery. AB - More surgeons are performing unilateral exploration for primary hyperparathyroidism (HPT) than ever before. This article reviews the factors that have led to the trend toward less invasive surgery. Discussion includes the history of unilateral exploration for HPT, the advent of magnetic resonance sestamibi imaging, and the development of intraoperative assays for parathyroid hormone. Results of minimally invasive techniques, including radio-guided parathyroidectomy, endoscopic parathyroidectomy, and outpatient parathyroidectomy, also are presented. PMID- 11059712 TI - Laparoscopic approach to adrenal and endocrine pancreatic tumors. AB - Laparoscopic adrenalectomy quickly has become the procedure of choice for benign adrenal lesions because it results in less pain, shorter hospital stay, comparable safety, and more patient satisfaction overall. The laparoscopic approach requires advanced laparoscopic surgical skills. Surgeons should be familiar with these techniques and the open approaches before attempting this procedure. When first learning the technique, small left-sided lesions are likely the easiest, and a more experienced surgeon should be present for the initial few cases; however, at this point, the laparoscopic approach to pancreatic endocrine tumors does not have a clear benefit, and it should be considered primarily investigational without clearly established benefits. PMID- 11059713 TI - Advanced laparoscopic gynecologic surgery. AB - What is the future for laparoscopy? Any procedure thought to be impossible to perform by laparoscopy or procedures that, based on conventional wisdom, should not be done laparoscopically are being performed or developed as the reader peruses this article. Technical advances in the endoscopic equipment and development of laparoscopic instruments have allowed for performance of sophisticated procedures with laparoscopic assistance. Appropriate laparoscopic skills allow surgeons to perform these procedures in a fashion nearly identical to an open procedure; however, modifications of historically proven techniques are controversial regarding the expenses generated, equipment necessary to perform the procedure, training necessary, and potential for complications. Has the obituary of laparotomy been written? The benefits of laparoscopically assisted or performed procedures are continuing to be analyzed. LAVH has been touted as a way to reduce the number of abdominal hysterectomies while increasing the number of vaginal hysterectomies. Therefore, indications for LAVH would ideally more resemble indications for abdominal hysterectomy than vaginal hysterectomy; however, LAVH does not seem to have increased the total number of vaginal hysterectomies. Conversely, the number of abdominal hysterectomies seems to be roughly the same, whereas the number of vaginal hysterectomies has decreased and the number of LAVHs has increased. Therefore, surgeons seem to be substituting LAVH for vaginal hysterectomy. Studies comparing laparoscopic Burch procedures and open Burch procedures are just now being reported. Many early reports described procedures that are not classic Burch colposuspensions. These changes make it impossible to assume that overall success and rate of complications are the same. The same can be said for techniques for correction of pelvic organ prolapse. Although laparoscopic performance and laparoscopic assistance are increasing in popularity, most cases are not handled in this way. Clearly, not every surgeon has embraced using the laparoscope to treat patients who would otherwise have undergone abdominal or vaginal surgery. PMID- 11059714 TI - Laparoscopy in urology. AB - Mention of all of the procedures in urology that have been attempted, or are being done, laparoscopically is beyond the scope of this article. The laparoscopic procedures outlined in this article are gaining increasing support as surgeons attempt to redefine gold standard minimally invasive therapies in the new millennium. Additional procedures, such as laparoscopic retroperitoneal lymph node dissections for low-stage, nonseminomatous germ cell testicular cancers and laparoscopic renal cryoablation of small renal cancers, are soon to be added to this list. As laparoscopic instrumentation and equipment continue to improve, it will become possible to explore even more procedures laparoscopically. Advances in imaging techniques, lasers, miniaturized robotics, and other areas may further define what is meant by the term minimal access surgery in the decades to follow. PMID- 11059715 TI - Laparoscopic lumbar interbody spinal fusion. AB - Laparoscopic ALIF is an evolving technique requiring the participation of a laparoscopic surgeon experienced in advanced laparoscopic techniques and knowledgeable in anterior lumbar spinal exposures. Initial enthusiasm for this technique was fostered by the development of interbody fusion devices and a method of exposing the anterior lumbar spine, which takes advantage of the ability of minimally invasive surgeries to improve exposure and visualization while minimizing collateral tissue damage and injury to healthy tissue. Preliminary studies have demonstrated laparoscopic ALIF feasibility. These same studies have been able to prove only minor advantages with the laparoscopic versus open technique using the current implants and bone grafting techniques for single-level disc disease. General acceptance of laparoscopic ALIF awaits further investigation. Reasons for a lack of general acceptance include the expense of the interbody fusion devices and laparoscopic equipment, the unfamiliarity of this advanced laparoscopic technique to spine and general surgeons, and the steep learning curve of the procedure. Intraoperative complications that arise are often severe, such as vascular injuries. Many skeptics appropriately believe that initial enthusiasm and zealousness must be tempered with scientific effort that provides data from long-term follow-up. For laparoscopic ALIF to gain general acceptance, randomized comparisons of laparoscopic ALIF to open ALIF and posterior lumbar spinal fusion and controlled studies with long-term follow-up documenting symptomatic outcome variables and spinal fusion rates must be completed. As new modalities are developed, minimally invasive techniques may facilitate their utility. The indications, procedures, and surgical principles of ALIF are unchanged, and physicians must not invent indications to justify the technique; however, eventually we may be able to redefine the indications to take full advantage of the endoscopic techniques and biological advances. PMID- 11059716 TI - Thoracoscopic esophagomyotomy for achalasia. AB - Achalasia is characterized by the absence of peristalsis in the distal two thirds of the esophagus, failure of receptive relaxation of the lower esophageal sphincter, and dysphagia to both solids and liquids. Diagnosis is confirmed by barium swallow, esophageal manometry, and flexible endoscopy. Treatment is based primarily on disruption of the lower esophageal sphincter, which can be achieved by forceful dilation of surgical esophagomyotomy. Esophagomyotomy produces relief of symptoms in more than 90% of patients. PMID- 11059717 TI - Video-assisted thoracic surgery for diseases within the mediastinum. AB - VATS and concepts of minimal access thoracic surgery have revitalized many aspects of general thoracic surgery, including the surgical approach to diseases and conditions of the mediastinum. Proven surgical options that have been shunned by patients and referring physicians because of the perceived morbidity of thoracotomy have been reconsidered with the emergence of these minimal access surgical options. Continued critical review of the accumulating experience in VATS techniques will refine the surgical indications for VATS and open thoracotomy. PMID- 11059718 TI - Thoracoscopic evaluation and treatment of thoracic trauma. AB - VATS has a diagnostic and therapeutic role in the treatment of patients with chest trauma, but the basic rule of safety over technology must be applied. It is an effective means for managing diaphragmatic injuries, hemothorax, empyemas, and persistent air leaks in selected hemodynamically stable patients. An overview of reported series (Table 1) demonstrates that VATS can be used successfully in the evaluation of patients with blunt and penetrating trauma. In appropriately selected cases, thoracoscopy can prove to be useful, with conversion to thoracotomy in only 10% of patients. Additional studies must be performed to determine any cost benefit compared with conventional therapy. PMID- 11059719 TI - Thoracoscopic evaluation and treatment of pulmonary disease. AB - VATS wedge resection and lobectomy can be performed with reasonable morbidity and mortality. A cautious approach is appropriate for VATS lobectomy with proper patient selection, and the completeness of the cancer surgery should not be compromised. Only surgeons with the VATS skills that allow them to perform complex procedures should perform the procedure. PMID- 11059720 TI - Suturing and knotting techniques for thoracoscopic cardiac surgery. AB - The preceding description of E-CABG may seem excessively detailed, even redundant, for trained cardiac surgeons; however, the authors' extensive experience with training surgeons on endoscopic techniques suggests that, despite a high level of proficiency and dexterity that a surgeon may possess in open surgery, becoming equally proficient and dexterous in the endoscopic environment is not simple. Participating in an in-depth, systematic endoscopic microvascular surgery training program in a laboratory setting is essential before applying the previously described E-CABG techniques in humans. The E-CABG procedure is one of the most challenging endoscopic techniques. Successful completion of this procedure requires that the surgeon be motivated to succeed and willing to invest the time and effort necessary to develop the new skills. Also critical is the avoidance of the temptation to use devices and systems that promise to obviate the need to bother with learning these difficult endoscopic skills. Long term results of the minimally invasive approach remain to be defined. However, some early studies of port-access procedures are encouraging. To date, a prospective randomized clinical trial comparing conventional LAD bypass to E-CABG has not been conducted. Although most investigators believe that long term patency of the IMA to the LAD using either technique should be the same, this is as yet unproven. Nonetheless, the adaption of endoscopic skills by the cardiac surgeon will further advance the evolution of this specialty. PMID- 11059721 TI - Coronary Surgery: off-pump and port access. AB - Attempts to minimize the invasiveness of cardiac surgery have focused on decreasing access trauma and eliminating cardiopulmonary bypass. The initial procedures, minimally invasive direct coronary artery bypass (MIDCAB, limited access beating heart) and port access (limited access arrested heart), have become niche procedures. Off-pump coronary artery bypass (OPCAB, median sternotomy beating heart) presently accounts for approximately 15% of all coronary bypass operations performed in the United States. Morbidity and cost appear to be decreased with these procedures. Feasibility trials of endoscopic coronary bypass surgery using robotic devices are underway in many centers. It is anticipated that over the next 5 years the alternative approaches to conventional coronary artery bypass surgery will continue to grow as methods of coronary revascularization. PMID- 11059722 TI - Reducing the trauma of congenital heart surgery. AB - While describing the circulatory system in De Moto Cordis, in 1628, William Harvey developed precepts for investigation, which could be modified slightly to guide the adoption of new technology and technique in the twenty-first century. Harvey might suggest (1) careful and accurate observation and description of a new technique, (2) a tentative explanation of how the technique improves on existing techniques, (3) a controlled testing of the hypothesis, and (4) conclusions based on the results of the experiments. Also, he might admonish surgery today, with its massively enhanced capabilities for information management, to rigorously test the validity of these conclusions with quantitative reasoning. In the future, precise measurement of the "trauma" of surgery, or even an individual surgeon, may be possible, and the long-term impact of a chest wall incision on a patient's self-esteem may be predictable. Absent such objective measures, justifications for "minimally invasive" deviations from conventional technique in surgery for CHD lack substance. Morbidity, mortality, and physiological endpoints will continue to form the foundation for therapeutic plans; however, the potential for emerging technology to reduce the trauma of these plans remains tantalizing. PMID- 11059723 TI - Minimally invasive surgical training solutions for the twenty-first century. AB - Despite the tremendous impact of laparoscopic cholecystectomy on the practice of surgery over the past 9 years, minimally invasive surgery faces many challenges that must be addressed. SAGES and the American College of Surgeons already have defined guidelines that, if properly implemented, could eliminate most of these challenges. Medical educators must formulate a detailed program as to how these guidelines can be widely deployed with acceptable effectiveness. The current educational philosophies and techniques will not ensure widespread access to a standardized program that would support the achievement of the goals set forth by major surgical governing bodies. Therefore, new educational strategies and techniques that are assisted with the integration of cost-effective technology are needed. Suggested solutions include the deployment of a standardized, objective-based skill-development program that has a large database to evaluate the progress of participants. Next, the Internet, with its ability to transfer content with the click of a mouse, will play an increasing role in distant education. Video and audio streaming techniques will allow the deployment of content previously shackled to a CD-ROM platform. CD-ROM interactive technology also can help in developing clinical judgment with innovative strategies, such as Objective-Based Clinical Competency Evaluation Scenarios. Telecommunications will fuse the components of a coordinated distant learning strategy. Also, telecommunications will allow the availability of new training capabilities in the form of teleproctoring and telementoring to hospitals, no matter what their size or location. All of these components combined enable the realization of a continuing education program in minimally invasive surgery that is readily available to hospitals worldwide. Last, institutions, resident training programs, and individual surgeons must commit the time to partake in these cutting-edge programs for challenges facing us to be completely eliminated. A high priority must be placed on the resolution of these issues. PMID- 11059724 TI - A decreased serum concentration of nitrite/nitrate correlates with an increased plasma concentration of lactate during and after major surgery. AB - A decrease in the production of nitric oxide (NO) due to surgical stress has been reported. We investigated whether this decrease in NO production was related to cytokine induction and/or other clinical parameters. We therefore measured the concentrations of serum nitrite/nitrate (a stable end product of NO), serum interleukin (IL)-6, and standard clinical parameters in 13 patients undergoing major upper abdominal and thoracoabdominal surgery at preanesthesia (PRE), 2h after a surgical incision (2H), at the end of surgery (END), and on the morning of postoperative days 1 (POD 1) and 3 (POD 3). The serum concentration of nitrite/nitrate was thus found to have significantly decreased at END, POD 1, and POD 3 compared with PRE. In addition, the serum nitrite/nitrate concentration correlated negatively with the plasma lactate level, and no relationship was observed between the serum nitrite/nitrate level and either the serum IL-6 level or any other clinical parameters. Our findings thus suggest that the decrease in the serum nitrite/nitrate level might therefore be related to tissue hypoperfusion both during and after major surgery. PMID- 11059725 TI - Changes in T-cell receptor subsets after cardiac surgery in children. AB - T cells are divided into two subsets, alphabeta and gammadelta, according to the T-cell receptor (TCR) expressed. Recent findings indicate that gammadelta T cells serve as the first defense against microbial pathogens, and represent a link between innate and acquired immunity. We conducted a study to investigate the changes in circulating TCR subsets after cardiac surgery in children. Blood samples from 24 children who underwent cardiac surgery with cardiopulmonary bypass (CPB) were collected serially to analyze TCR subsets by flow cytometry. The alphabeta T cells reached a nadir on postoperative day (POD) 1, but recovered to pre-CPB levels on POD 3. On the other hand, the gammadelta T cells decreased after CPB and did not recover to pre-CPB levels even after POD 7. The alphabeta/gammadelta T-cell ratio was increased after POD 3. In children, gammadelta T cells recover more slowly than alphabeta T cells after cardiac surgery. These changes in TCR subsets may contribute to postoperative immunosuppression. PMID- 11059726 TI - Analysis of the anatomic changes in the thoracic cage after a lung resection using magnetic resonance imaging. AB - The thoracic cage after a lung resection is filled by the remaining lobes, the elevated diaphragm, the diminished thoracic cage, and by mediastinal shifting. The changes in the thorax after a lung resection were quantified using magnetic resonance imaging. The study group consisted of 39 patients who had undergone a lobectomy, four who had undergone a pneumonectomy, and 14 controls. The left ventricular angle, ascending aortic angle, mediastinal shift, longitudinal length of the thoracic cage, the distance between the thoracic apex and the level of the aortic valve, and diaphragmatic elevation were all measured. After a right lower lobectomy, the mediastinum shifted more rightward than after a right upper lobectomy. The diaphragm became more greatly elevated after a right upper lobectomy than after a right lower lobectomy. When a chest wall resection was added to a right upper lobectomy, the mediastinal anatomical changes decreased. After a left upper lobectomy, the degree of mediastinal shifting was greater than after a left lower lobectomy. A left upper lobectomy shifted the mediastinum at the level of the right atrium. This method is easily reproducible and was found to be effective for quantifying the changes in the thorax after a lung resection. PMID- 11059727 TI - Key factors influencing bowel function after ileal W-pouch anal anastomosis: a spectral analysis of W-pouch motor activity. AB - Restorative proctocolectomy with ileal pouch anal anastomosis (IPAA) has become the standard surgical procedure for ulcerative colitis (UC). The purpose of this study was to determine which factors are important to achieve good anal continence after IPAA in terms of the motor activity and pressure-volume relationship. A total of 17 patients with UC who underwent IPAA were evaluated. The internal ileal pouch pressure was transanally measured with and without volume-loading of the pouch which induces the urge to evacuate. The maximum tolerable volume (MTV), first urge volume (FUV), and ileal pouch compliance were calculated and the internal ileal pouch pressure records were subjected to spectral analysis for intensive evaluation of the intraluminal pressure waves. The FUV, correlation of the compliance of the FUV with MTV, and the remaining volume up to the MTV (RVMTV) were analyzed. Compliance of the FUV was significantly correlated with the RVMTV (r = 0.736, P < 0.01). The frequency of the phasic waves in the pouch decreased with length of follow up, reflecting improved function (r = -0.588, P < 0.05). The findings of this intensive analysis of manometric measurement indicate that the key factors in postoperative pouch function are RVMTV and the frequency of phasic waves in the W-pouch. PMID- 11059728 TI - The clinical significance of lymph node metastases in patients undergoing surgery for hepatocellular carcinoma. AB - The frequency of lymph node (LN) metastasis in patients undergoing surgery for hepatocellular carcinoma (HCC) has rarely been studied. We evaluated the clinicopathologic characteristics and outcomes of six patients with nodal metastases from HCC among a total of 504 patients who underwent hepatic resection for HCC in our department over a 16-year period. The nodal metastases were diagnosed preoperatively in two patients. The average diameter of the resected tumors was 7.8 cm and all were confirmed as poorly differentiated HCC. All of the six patients had intrahepatic metastatic nodules and five also had portal vein invasion. One patient underwent limited resection, and the other five underwent bisegmentectomy. All of the regional LNs were removed in one patient, while only enlarged LNs were removed in the other five. One patient died of postoperative liver failure and the others all died later of intrahepatic or nodal recurrence. Our findings suggest that the prognosis of patients with nodal metastasis from HCC is generally poor, even if hepatic resection with regional LN dissection is performed. PMID- 11059729 TI - The effect of pentoxifylline on the healing of intestinal anastomosis in rats with experimental obstructive jaundice. AB - The aims of this study were (1) to investigate the effect of experimental obstructive jaundice on the healing of intestinal anastomosis, and (2) to investigate the effect of pentoxifylline on the healing of intestinal anastomosis in rats with obstructive jaundice. Obstructive jaundice was induced in rats by the ligation and division of the common bile duct. Four days after this operation, either pentoxifylline or isotonic saline solution was administered intraperitoneally to these jaundiced rats and controls, and then intestinal anastomosis was performed. The concentrations of serum tumor necrosis factor alpha (TNF-alpha) and serum triglyceride of jaundiced and nonjaundiced rats were measured, and the quality of healing was evaluated by measuring the bursting pressure and hydroxyproline content of the anastomoses on the fifth and tenth days of anastomotic healing. Obstructive jaundice resulted in an impaired wound healing of the intestinal anastomosis in the rats. The administration of pentoxifylline to the jaundiced rats resulted in better anastomotic wound healing. The beneficial effects of pentoxifylline on anastomotic healing in rats with obstructive jaundice was attributed to its inhibitor effect on the endotoxin induced TNF-alpha release from macrophages and monocytes, and the stabilizing effect on the neutrophils. PMID- 11059730 TI - The cytotoxic effects of bile acids in crude bile on human pancreatic cancer cell lines. AB - Pancreatic cancer frequently causes extrahepatic cholestasis. To identify the direct effects of bile acids in jaundiced serum on pancreatic cancer, the proliferation of PANC-1 and MIA PaCa-2 cells as well as the ultrastructural alteration of PANC-1 cells cultured in crude bile modified media were studied. The growth of these cells in the RPMI-1640 media with or without 1%, 2%, and 4% of the refined crude bile was assessed after 48 and 96 h of incubation. The ultrastructure of PANC-1 cells was investigated by scanning and transmission electron microscopy after 24 and 48 h of incubation. The proliferation of both cell lines in the bile-treated media was greatly inhibited. The inhibitory rates of bile on PANC-1 ranged from 24.1% +/- 3.3% to 66.9% +/- 6.6% (P < 0.01) and those on MIA PaCa-2 ranged from 16.7% +/- 3.8% to 50.7% +/- 5.5%. (P < 0.01). When the bile-added media were replaced, the cells were able to restore their proliferating ability. The PANC-1 cells incubated in the bile-supplied media indicated that the mirovilli, mitochondria, and other organelles had thus been injured. These results suggest that bile acids appear to inhibit the proliferation of PANC-1 and MIA PaCa-2 cells, and the probable inhibitory mechanism is mainly considered to be due to the cytotoxicity of such bile acids. PMID- 11059731 TI - The likely transformation of papillary thyroid carcinoma into anaplastic carcinoma during postoperative radioactive iodine-131 therapy: report of a case. AB - We report herein a case of papillary carcinoma which appeared to transform into anaplastic carcinoma during postoperative radioactive iodine-131 (131I) therapy. A 67-year-old man who was diagnosed as having papillary thyroid carcinoma with bilateral neck lymph node involvement and multiple lung metastases underwent total thyroidectomy prior to 131I therapy. Immediately after a second course of 131I therapy, the patient complained of right neck pain and swelling, and a biopsy of the swollen neck lymph node was taken. Histologic examination of this biopsy specimen revealed anaplastic carcinoma. With p53 immunohistochemical staining, both the primary tumor and the biopsy specimen were positive. We speculate that first, some DNA damage in tumor cells was induced by the initial 131I therapy, but neither DNA repair nor cell apoptosis occurred because the p53 gene was already mutated; then further DNA damage was induced by the second 131I therapy, leading to anaplastic transformation. PMID- 11059732 TI - Delayed infection of a lymphocele following mastectomy with immediate breast reconstruction: report of a case. AB - We report herein a rare case of delayed infection of a lymphocele following mastectomy with immediate breast reconstruction. A 38-year-old woman presented to our hospital 7 months after undergoing a left-modified radical mastectomy with an immediate breast reconstruction, following the sudden development of a giant mass in the left thoracoabdominal region as well as a high fever and shivering. Ultrasonography and a computed tomographic scan revealed massive fluid retention extending from the left axilla to the lower abdominal region. Puncture drainage was performed three times and the injection of an antibiotic directly into the cyst resulted in resolution of the fluid. This massive retraction of fluid was considered to have resulted from a delayed infection of an axillary lymphocele. PMID- 11059733 TI - Completion pneumonectomy of the residual left lung to treat lung cancer in a patient with hemophilia A: report of a case. AB - Hemophilia A is a sex-linked recessive hereditary disease that is relatively rare and the number of patients with this disorder who undergo major surgery is limited. Although replenishing coagulation factors can allow hemophiliac patients to undergo similar surgery to that performed for patients without hemophilia, there have been few reports on major surgery and none on the resection of lung cancer in patients with hemophilia A. We recently performed completion pneumonectomy of the left lung in a 70-year-old man with hemophilia A, for squamous cell carcinoma in the residual left lung. The administration of a recombinant DNA coagulation factor VIII preparation allowed this operation to be successfully carried out. This case serves to demonstrate that the recombinant DNA coagulation factor VIII preparation described may enable us to safely perform major surgery on hemophiliac patients, since there is no risk of viral infection or any other adverse effects, such as deterioration of immunocompetence or hemolysis, which are occasionally encountered with human plasma-derived preparations. PMID- 11059734 TI - Isolated gastric tuberculosis presenting as massive hematemesis: report of a case. AB - Tuberculous involvement of the stomach is rare. We report herein the unusual case of a 25-year-old man in whom a benign gastric ulcer was found along the lesser curvature after he presented with massive upper gastrointestinal bleeding. Histopathological examination helped to confirm a diagnosis of tuberculosis. The granulomas typical of tuberculosis were caseation with epithelioid and giant cells. The patient was successfully treated by a combination of appropriate surgical therapy and prompt institution of antituberculosis medication. PMID- 11059735 TI - Development of an inveterate gastroduodenal ulcer caused by antral G-cell hyperplasia of the stomach (pseudo-Zollinger-Ellison Syndrome): report of a case. AB - We describe herein the case of a 54-year-old Japanese woman in whom an inveterate peptic ulcer developed in association with pseudo-Zollinger-Ellison Syndrome (pseudo-ZES). The patient presented with weight loss and abdominal distension caused by antral and duodenal stenosis due to an inveterate peptic ulcer. Her serum gastrin level was very high; however, no evidence of a gastrinoma or carcinoid tumor was detected by preoperative examinations or surgery. A total gastrectomy and double-tract reconstruction was performed, and pathological examination revealed a gastric ulcer (UL-IV) with no histopathological evidence of a neoplasm. Immunohistochemical staining showed an obvious increase in the number of endocrine cells that were positive for chromogranin A, and marked G cell hyperplasia was observed in the antral mucosa. Furthermore, the number of enterochromaffin-like cells was remarkably high. From the results of the immunohistochemical examination, the patient was diagnosed as having hypergastrinemia due to antral G-cell hyperplasia. Postoperatively, the patient's serum gastrin level fell rapidly to within the normal range, her nutritional status improved, and her weight increased by about 10 kg within 1 year. PMID- 11059736 TI - Adenocarcinoma of the ileum producing carbohydrate antigen 19-9: report of a case. AB - We report herein the case of an 81-year-old woman found to have small intestinal carcinoma producing carbohydrate antigen (CA)19-9, in whom recurrence on the abdominal wall was strongly suspected 4 months after resection. She presented to our hospital with acute abdominal pain with severe anemia. Marked serum elevation of CA19-9 to 164.8 U/ml suggested a progression to malignancy. A fluorography using an ileus tube revealed an abnormal mucosal pattern. An exploratory laparotomy showed an incomplete annular constrictive Borrmann type 2 tumor, located approximately 190 cm from Treitz's ligament, without any signs of peritoneal or hepatic metastases. Histological examination confirmed a diagnosis of papillotubular adenocarcinoma without metastases of the regional lymph nodes. CA19-9 antigenicity was detected in the cytoplasm and on the surface of the cancer cells, using the monoclonal CA19-9 antibody, NS19-9. In this report, we demonstrate the CA19-9 productivity and distribution of the cancer tissues in relation to their prognosis. PMID- 11059737 TI - Successful resection of a liver metastasis from gastric leiomyoblastoma: report of a case. AB - A 20-year-old woman was referred to our hospital for detailed investigation of a gastric submucosal tumor. A leiomyoma was preoperatively diagnosed and laparoscopic-assisted enucleation was performed. The resected tumor was 4 x 3 x 1.5 cm in size and postoperative histological examination identified it as a gastric leiomyoblastoma. Therefore, a secondary resection in the form of a distal gastrectomy was carried out. No tumor cells were found in the gastric specimen or in the lymph nodes; however, 5 months after the operation, an abdominal computed tomography scan revealed a recurrence in the liver, and she was readmitted for further examinations. The lesion was diagnosed as a single liver metastasis from the gastric leiomyoblastoma and successfully resected. The histopathological findings of the liver tumor resembled those of the primary gastric tumor. Her postoperative course was uneventful and she has been well, without any evidence of recurrence, to date. Only 12 other cases of leiomyoblastoma of the stomach with liver metastasis have been reported in Japan, all of which were associated with a very poor prognosis. Therefore, patients with this unusual disease entity should be carefully followed up after resection of the primary tumor. PMID- 11059738 TI - Insulinoma occurring in association with fatty replacement of unknown etiology in the pancreas: report of a case. AB - A 66-year-old woman with a 10-year-history of diabetes mellitus was admitted to our hospital for investigation of several recent attacks of hypoglycemia. Her fasting blood glucose level was very low, at 30-40 mg/dl, and abdominal ultrasonography and computed tomography revealed a tumor in the pancreatic tail with fatty changes. Endoscopic retrograde cholangiopancreatography revealed absence of the main pancreatic duct from the body to tail of the pancreas. Abdominal angiography showed a hypervascular tumor stain in the pancreas, and percutaneous transhepatic portal vein sampling demonstrated a step-up of immunoreactive insulin levels in the splenic vein. Based on these clinical findings, we made a preoperative diagnosis of an insulinoma accompanied by fatty changes in the pancreatic body and tail. During laparotomy for the insulinoma, fat tissue was identified in the anatomic location of the pancreatic body and tail, and resected. Pathological examination of the resected specimen revealed a number of Langerhans islets in the adipose tissue, and an islet cell tumor with fatty replacement of the pancreatic tissue around the tumor. The insulinoma was found not to have caused obstruction of the main pancreatic duct. We present herein a rare case of an insulinoma that developed in the pancreas, and was associated with fatty replacement of unknown etiology. PMID- 11059739 TI - Inflammatory pseudotumor of the spleen: report of a case. AB - A case of a 45-year-old Japanese man with a splenic inflammatory pseudotumor is described. This benign lesion is rarely reported in the world literature. We preoperatively could not rule out the possibility of a malignant neoplasm, due to the fact that the tumor had grown in size after a 2-year observation. However, after performing a splenectomy, a histological examination of the mass revealed an inflammatory process. Inflammatory pseudotumors often pose diagnostic difficulties because the clinical and radiological findings tend to suggest a malignancy. The clinical and pathological features of such previously reported cases are also reviewed. PMID- 11059740 TI - Musculoskeletal and adipose tissue hydatidosis based on the iatrogenic spreading of cystic fluid during surgery: report of a case. AB - Hydatidosis or echinococcosis is a parasitic disease caused by Echinococcus granulosus or E. multilocularis, which forms cysts in the liver and lung after penetrating the duodenal mucosa and entering the portal circulation. The liver and lung act as a filter but some embryos enter the general circulation and disseminate throughout the body. Musculoskeletal involvement is a rare manifestation of hydatidosis, which is usually reported to affect a single muscle. We report here a rare case of a 68-year-old man with widespread hydatidosis of the retroperitoneum and the subcutaneous adipose tissue, and with multiple muscle involvement in the absence of liver, lung, and spleen involvement. The patient underwent surgical excision of a subcutaneous hydatid cyst 7 years earlier. It is likely that the large dissemination of parasites resulted from accidental rupture of the primary focus during surgery with consequent release and spreading of scolices via lymphatics. PMID- 11059741 TI - Abdominal cocoon: report of a case. AB - The abdominal cocoon is a rare cause of intestinal obstruction most often found in adolescent girls from tropical and subtropical countries. It is characterized by a thick fibrotic sac covering the small bowel partially or completely, the etiology of which is unknown. A correct diagnosis is not often made preoperatively; however, following simple surgical release of the entrapped bowel, these patients usually do well. We report herein our experience of a case of abdominal cocoon with a brief review of the medical literature on this unusual disease entity. PMID- 11059742 TI - Internal hernia with triple hiatus of congenital origin: report of a case. AB - We report herein the case of a 65-year-old woman who was referred to our department with prolonged ileus symptoms despite conservative therapy. A plain abdominal radiograph showed intestinal gas shadows with an air-fluid level in the lesser curvature of the stomach. As no improvement was achieved by the insertion of a short tube, a long tube was inserted. A loop formation of the long tube in the subphrenic region was detected on an abdominal radiograph, and an enterogram showed an interruption in the ileum in the lower abdomen. The patient was diagnosed as having an adhesional ileus and a strangulated ileus due to a lesser sac hernia. A laparotomy was performed which revealed that the small intestine had herniated into the lesser sac space through a hiatus of Treitz' fossa and a hiatus in the transverse mesocolon. Furthermore, part of the small intestine had herniated through an omental hiatus. The herniated intestine was manually reduced and the hiatus was closed. However, as the right ovary was found to have adhered to the ileum and stenosis was seen, we were forced to perform partial resection of the ileum. Considering that this patient had no history of laparotomy in the upper abdomen, abdominal injury, or acute abdomen, it was surmised that the three abnormal hiatuses were congenital. PMID- 11059743 TI - Hepatic resection using the harmonic scalpel. AB - We describe herein our technique of performing extensive resection of the liver by blunt dissection in combination with excision using a harmonic scalpel. A ball coagulator was inserted at 3-cm intervals along the proposed cutting line in the liver, and the liver parenchyma between these holes was then cut using coagulation shears. Regardless of the condition of the liver, good coagulation and cutting were achieved using the harmonic scalpel without vascular occlusion when dividing the shallow layer of the liver, and no complications in association with the harmonic scalpel, such as postoperative bleeding, bile leakage, or abscess formation at the cut margins, occurred. In the deep layer below the main trunk of the hepatic vein, blunt dissection was used, since it was difficult to achieve sufficient control of bleeding from large vessels using the harmonic scalpel alone. Therefore, when used in combination with other techniques, the harmonic scalpel appears to be an effective device for liver surgery that minimizes bleeding and decreases the vascular clamping time. PMID- 11059744 TI - An improved method for distinguishing the intersegmental plane of the lung. AB - An improved technique for distinguishing the intersegmental plane of the lung was developed as follows. After the involved bronchus is identified, the lobe is inflated and the segmental bronchus is then tied to maintain gas inside of the segments that will be removed, and thereafter is severed at a point proximal to the tie. When almost done closing the stump, a line will develop between the deflated and the inflated area, which represents the intersegmental plane to be operated on. This technique is therefore completely different from the technique described in textbooks, in which the preserved segment is kept inflated while the resected one is kept deflated. Once the line develops, one can operate just on the line using either electrocautery under adequate tension or staples between the collapsed and inflated segments. The cutting surface is so close to the real intersegmental plane that the amounts of air leak and bleeding are negligible. PMID- 11059745 TI - Celecoxib inhibits N-butyl-N-(4-hydroxybutyl)-nitrosamine-induced urinary bladder cancers in male B6D2F1 mice and female Fischer-344 rats. AB - Epidemiological studies have shown that nonsteroidal anti-inflammatory drugs (NSAIDs) may have a role in the prevention of human cancers. A number of preclinical studies have also suggested that inhibition of cyclooxygenase (COX) with NSAIDs has an anticancer effect in animal models of colon, urinary bladder, skin, and breast. In these studies, we evaluated the COX-2 inhibitor celecoxib in two rodent models of urinary bladder cancer. Male B6D2F1 mice treated with N butyl-N-(4-hydroxybutyl)-nitrosamine (OH-BBN) developed transitional and squamous cell urinary bladder cancers, many of which grew rapidly and caused substantial morbidity that required sacrifice of the mice. Groups of mice received various daily doses of celecoxib in the diet (1250, 500, or 200 mg/kg of diet) beginning 7 days before the initiation of 12 weekly doses of OH-BBN. Mice were checked weekly for the presence of palpable urinary bladder masses. The study was terminated at 8 months following the initial treatment with OH-BBN. The percentage of mice with large palpable bladder lesions, which necessitated sacrifice of the mice, was 40% in the OH-BBN control group. In contrast, only 10% of all celecoxib-treated mice required sacrifice before the scheduled termination of the experiment, implying that all three doses of celecoxib inhibited the formation of large palpable lesions. Celecoxib did not significantly alter the incidence of preneoplastic bladder lesions, but did dose-dependently decrease the total number of urinary bladder cancers/mouse, palpable plus microscopic, by 77, 57, and 43% at dosages of 1250, 500, and 200 mg of celecoxib/kg of diet, respectively. In the second model, female Fischer-344 rats were administered OH BBN twice/week for a period of 8 weeks. After 8 months, all rats developed preneoplastic lesions, whereas roughly 60% of the rats developed relatively small urinary bladder cancers. Rats were treated continually with celecoxib in the diet (500 or 1000 mg/kg of diet) beginning either 1 week prior to the initial OH-BBN treatment or beginning 1 week following the last OH-BBN treatment. Neither celecoxib treatment regimen significantly altered the number of preneoplastic lesions. Whereas celecoxib treatment initiated prior to OH-BBN administration decreased cancer incidence roughly 65%, celecoxib treatment initiated beginning 1 week after the last dose of OH-BBN profoundly decreased cancer incidence (>95%). Celecoxib did not alter the body weights of the mice or rats, or cause other signs of toxicity at any of the doses studied. Taken together these results demonstrate that: (a) celecoxib effectively inhibits tumor growth and enhances survival in the mouse model of urinary bladder cancer; and (b) celecoxib profoundly inhibits development of urinary bladder cancers in the rat model even when administered following the last dose of OH-BBN. Clinical trials will be necessary to determine whether COX-2 inhibitors will provide a clinical benefit in human bladder cancer. PMID- 11059746 TI - Expression and function of CYR61, an angiogenic factor, in breast cancer cell lines and tumor biopsies. AB - We have previously shown that expression of heregulin (HRG) is closely correlated with breast cancer progression. We have subsequently isolated Cyr61, a ligand for the alpha(v)beta3 integrin that is differentially expressed in HRG-positive cells, and have shown that it is expressed in all of the invasive and metastatic breast cancer cell lines tested. Preliminary evaluation of Cyr61 expression in breast tumor biopsies revealed expression of Cyr61 in about 30% of invasive breast carcinomas. Significantly, we demonstrated that Cyr61 is a downstream effector of HRG action, because a Cyr61-neutralizing antibody abolished the ability of HRG-expressing cells to migrate in vitro. Furthermore, we have shown that HRG-expressing cells denote higher levels of alpha(v)beta3 expression, and we have established that Cyr61 action is mediated, at least in part, through its receptor alpha(v)beta3, because a functional blocking antibody of the alpha(v)beta3 blocked the Matrigel outgrowth of HRG-expressing cells. These results strongly suggest that Cyr61 is necessary for HRG-mediated chemomigration and that Cyr61 plays a functional role in breast cancer progression, possibly through its interactions with the alpha(v)beta3 receptor. PMID- 11059747 TI - Chromosomal instability in unirradiated cells induced in vivo by a bystander effect of ionizing radiation. AB - Using a bone marrow transplantation protocol in which we transplanted a mixture of irradiated and nonirradiated bone marrow cells that were distinguishable by a cytogenetic marker, we have demonstrated chromosomal instability in the progeny of nonirradiated hemopoietic stem cells. This first demonstration of a link between a bystander effect of ionizing radiation and the induction of genomic instability in vivo clearly poses a major challenge to current views of the mechanisms of radiation-induced DNA damage with mechanistic implications for the health consequences of radiation exposure particularly in the context of the induction of malignancy. PMID- 11059748 TI - A variant within the DNA repair gene XRCC3 is associated with the development of melanoma skin cancer. AB - Exposure to UV radiation is a major risk factor for the development of malignant melanoma. DNA damage caused by UV radiation is thought to play a major role in carcinogenesis induction. Multiprotein pathways involved in repairing UV-DNA damage are the base excision, the nucleotide excision, and the homologous double stranded DNA repair pathways. This study used a sequence-specific primer PCR (PCR SSP) genotyping method to investigate the association between polymorphisms in DNA repair genes from these pathways with the development of malignant melanoma. The patient cohort was comprised of 125 individuals with malignant melanoma with lesions or staging suggesting a high risk of relapse or metastatic disease. The control population consisted of 211 individuals. We found the presence of a T allele in exon 7 (position 18067) of the XRCC3 gene was significantly associated with melanoma development (P = 0.004; odds ratio, 2.36; relative risk, 1.74). This gene codes for a protein involved in the homologous pathway of double stranded DNA repair, thought to repair chromosomal fragmentation, translocations, and deletions. These results may provide further insights into the pathogenesis and the mechanism of UV-radiation induced carcinogenesis as well as having a role in prevention. PMID- 11059749 TI - Silibinin up-regulates insulin-like growth factor-binding protein 3 expression and inhibits proliferation of androgen-independent prostate cancer cells. AB - Silibinin, a naturally occurring flavonoid antioxidant found in the milk thistle, has recently been shown to have potent antiproliferative effects against various malignant cell lines, but the underlying mechanism of action remains to be elucidated. We investigated the effect of silibinin on androgen-independent prostate cancer PC-3 cells. At pharmacologically achievable silibinin concentrations (0.02-20 microM), we observed increased insulin-like growth factor binding protein 3 (IGFBP-3) accumulation in PC-3 cell conditioned medium and a dose-dependent increase of IGFBP-3 mRNA abundance with a 9-fold increase over baseline at 20 microM silibinin. An IGFBP-3 antisense oligodeoxynucleotide that attenuated silibinin-induced IGFBP-3 gene expression and protein accumulation reduced the antiproliferative action of silibinin. We also observed that silibinin reduced insulin receptor substrate 1 tyrosine phosphorylation, indicating an inhibitory effect on the insulin-like growth factor I receptor mediated signaling pathway. These results suggest a novel mechanism by which silibinin acts as an antiproliferative agent and justify further work to investigate potential use of this compound or its derivatives in prostate cancer treatment and prevention. PMID- 11059750 TI - Association between survival after treatment for breast cancer and glutathione S transferase P1 Ile105Val polymorphism. AB - A glutathione S-transferase (GST) P1 polymorphism results in an amino acid substitution, Ile105Val; the Val-containing enzyme has reduced activity toward alkylating agents. Cancer patients with the variant enzyme may differ in removal of treatment agents and in outcomes of therapy. We evaluated survival according to GSTP1 genotype among women (n = 240) treated for breast cancer. Women with the low-activity Val/Val genotype had better survival. Compared with Ile/Ile, hazard ratios for overall survival were 0.8 (95% confidence interval, 0.5-1.3) for Ile/Val and 0.3 (95% confidence interval, 0.1-1.0) for Val/Val (P for trend = 0.04). Inherited metabolic variability may influence treatment outcomes. PMID- 11059751 TI - Scavenging of reactive oxygen species leads to diminished peritoneal tumor recurrence. AB - Previously, we demonstrated that RBCs inhibit the recurrence of perioperatively spilled tumor cells. The aim of this study was to identify on which RBC component(s) the inhibitory effect is based. By using a cell-seeding model in rats, the effect of RBC-related antioxidant scavengers [hemoglobin, catalase, and superoxide dismutase (SOD)] on peritoneal tumor recurrence was investigated. i.p. injection of hemoglobin caused 45% more tumor load (P < 0.0001). At least 40% inhibition of tumor recurrence was achieved with the use of catalase or SOD (P < 0.05). Combining SOD and catalase did not lead to additional inhibition of tumor recurrence. Inhibition of the overwhelming oxidative potential after surgical peritoneal trauma with the use of scavengers may lead to interesting new approaches for diminishing peritoneal tumor recurrence. PMID- 11059752 TI - Hyperinducibility of hypoxia-responsive genes without p53/p21-dependent checkpoint in aggressive prostate cancer. AB - Hypoxia limits tumor growth but selects for higher metastatic potential. We tested the functional activity of hypoxia-inducible factor-1 (HIF-1) in prostate cell lines ranging from normal epithelial cells (PrEC), hormone-dependent LNCaP, hormone-independent DU145, PC-3 to highly metastatic PC-3M cancer cell lines. We found that HIF-1-stimulated transcription was the lowest in PrEC and LNCaP cells and the highest in PC-3M cells. The induction by hypoxia of the HIF-1 dependent genes Cap43 and GAPDH was the highest in the most aggressive PC-3M cancer cells. Because these advanced prostate cancer cell lines have lost p53 function, this further shifts a balance from p53 to HIF-1 transcriptional regulation, and a high ratio of HIF-1-dependent:p53-dependent transcription was a marker of the advanced malignant phenotype. Transient transfection of HIF-1alpha expression vector induced transcription from p21 promoter construct in prostate cancer cell lines. Furthermore, hypoxia slightly induced p21 mRNA in these cells. However, neither expression of p21 nor hypoxia caused growth arrest in PC-3M cells. Therefore, high inducibility of HIF-1-dependent genes, loss of p53 functions with high ratio of HIF-1-dependent:p53-dependent transcription, and loss of sensitivity to p21 inhibition is a part of hypoxic phenotype associated with aggressive cancer behavior. PMID- 11059753 TI - Alcohol stimulates estrogen receptor signaling in human breast cancer cell lines. AB - Epidemiological studies suggest that moderate alcohol consumption increases the risk of breast cancer, and that alcohol combined with estrogen replacement therapy may synergistically enhance the risk. However, the mechanism(s) of alcohol-induced mammary cancer is unknown. In human breast cancer cell lines, we found that ethanol (EtOH) caused a dose-dependent increase of up to 10- to 15 fold in the transcriptional activity of the liganded estrogen receptor (ER alpha), but did not activate the nonliganded receptor. Significant stimulation of ER-alpha activity was observed at EtOH concentrations comparable with or less than blood alcohol levels associated with intoxication and at doses below the threshold for in vitro cytotoxicity. These findings may be explained, in part, by an EtOH-induced down-regulation of the expression of BRCA1, a potent inhibitor of ER-alpha activity, and, in part, by a modest increase in the ER-alpha levels. Our findings suggest that inactivation of BRCA1 and increased estrogen-responsiveness might contribute to alcohol-induced breast cancer. PMID- 11059754 TI - Somatic mutations of the CD95 gene in Hodgkin and Reed-Sternberg cells. AB - Hodgkin and Reed-Sternberg (H/RS) cells in classical Hodgkin's disease (cHD) are thought to be derived from preapoptotic germinal center B cells. However, little is known about the transforming events rescuing the precursor of the H/RS cells from apoptosis. Given the importance of CD95 (Apo-1/Fas)-mediated apoptosis for negative selection within the germinal center, single micromanipulated H/RS cells from 10 cases of cHD were analyzed for somatic mutations within the CD95 gene. Three clonal mutations within the 5' regions were amplified from single H/RS cells in one case. From H/RS cells of another case, two mutations within the last exon coding for the death domain were detected. About half of these H/RS cells carried a monoallelic stop-codon; the remaining tumor cells harbored a monoallelic replacement mutation. Both mutations likely impair CD95 function. Because all these H/RS cells also bear clonal mutations inactivating the IkappaB alpha gene, the IkappaB alpha mutations occurred earlier than those of the CD95 gene in the sequence of transforming events leading to cHD. In conclusion, somatic mutations of the CD95 gene occur in a fraction of cHD cases and may favor the escape of the precursor of the H/RS clone from apoptosis. PMID- 11059755 TI - BCL-6 mutations are associated with immunoglobulin variable heavy chain mutations in B-cell chronic lymphocytic leukemia. AB - The cell of origin of B-cell chronic lymphocytic leukemia (B-CLL) is still uncertain. Recent studies have indicated that a fraction of B-CLL displays somatically mutated immunoglobulin variable heavy chain (IgV(H)) genes, which suggests an origin from a post-germinal center (GC) B cell. It has been shown that the 5' noncoding region of the BCL-6 proto-oncogene is affected by mutations in normal GC B-lymphocytes and in lymphoid malignancies displaying GC/post-GC phenotype. To further explore the cellular origin of B-CLL, we have analyzed 34 cases for mutations in the BCL-6 5' noncoding region and in the IgV(H) genes. We found somatically mutated IgV(H) genes in 24 (73%) of 33 samples (average frequency, 6.5 x 10(-2)/bp) and BCL-6 mutations in 8 (24%) of 34 cases (average frequency, 0.14 x 10(-2)/bp in the mutated cases). The occurrence of BCL-6 mutations was restricted to those cases displaying IgV(H) mutations. Analysis of BCL-6 protein expression as a marker of GC phenotype showed that, regardless of the presence of IgV(H) or BCL-6 mutations, B-CLLs express BCL-6 at levels clearly below those found in normal or transformed GC B cells. These results indicate that a subset of B-CLL derives from a cell that has been exposed to the somatic hypermutation mechanism and support the hypothesis that BCL-6 mutations result from the same process that targets immunoglobulin genes. PMID- 11059756 TI - Melanoma-targeting properties of (99m)technetium-labeled cyclic alpha-melanocyte stimulating hormone peptide analogues. AB - Preliminary reports have demonstrated that (99m)technetium (Tc)-labeled cyclic [Cys(3,4,10), D-Phe7]alpha-MSH(3-13) (CCMSH) exhibits high tumor uptake and retention values in a murine melanoma mouse model. In this report, the tumor targeting mechanism of 99mTc-CCMSH was studied and compared with four other radiolabeled alpha-melanocyte stimulating hormone (alpha-MSH) peptide analogues: 125I-(Tyr2)-[Nle4, D-Phe7]alpha-MSH [125I-(Tyr2)-NDP]; 99mTc-CGCG-NDP; 99mTc Gly11-CCMSH; and 99mTc-Nle11-CCMSH. In vitro receptor binding, internalization, and cellular retention of radiolabeled alpha-MSH analogues in B16/F1 murine cell line demonstrated that >70% of the receptor-bound radiolabeled analogues were internalized together with the receptor. Ninety % of the internalized 125I-(Tyr2) NDP, whereas only 36% of internalized 99mTc-CCMSH, was released from the cells into the medium during a 4-h incubation at 37 degrees C. Two mouse models, C57 mice and severe combined immunodeficient (Scid) mice, inoculated s.c. with B16/F1 murine and TXM-13 human melanoma cells were used for the in vivo studies. Tumor uptake values of 11.32 and 2.39 [% injected dose (ID)/g] for 99mTc-CCMSH at 4 h after injection, resulted in an uptake ratio of tumor:blood of 39.0 and 11.5 in murine melanoma-C57 and human melanoma-Scid mouse models, respectively. Two strategies for decreasing the nonspecific kidney uptake of 99mTc-CCMSH, substitution of Lys11 in CCMSH with Gly11 or Nle11, and lysine coinjection, were evaluated. The biodistribution data for the modified peptides showed that Lys11 replacement dramatically decreased the kidney uptake, whereas the tumor uptakes of 99mTc-Nle11- and 99mTc-Gly11-CCMSH were significantly lower than that of 99mTc CCMSH. Lysine coinjection significantly decreased the kidney uptake (e.g., from 14.6% ID/g to 4.5% ID/g at 4 h after injection in murine melanoma-C57 mice) without significantly changing the value of tumor uptake of 99mTc-CCMSH. In conclusion, the compact cyclic structure of 99mTc-CCMSH, its resistance to degradation, and its enhanced intracellular retention are the major contributing factors to the superior in vivo tumor targeting properties of 99mTc-CCMSH. Lys11 residue in 99mTc-CCMSH is critical to the tumor targeting in vivo, and lysine coinjection rather than lysine replacement can significantly decrease the nonspecific renal radioactivity accumulation without impeding the high melanoma targeting properties of 99Tc-CCMSH. The metal-cyclized CCMSH molecule displays excellent potential for the development of melanoma-specific diagnostic and therapeutic agents. PMID- 11059757 TI - Down-regulation of X-linked inhibitor of apoptosis protein induces apoptosis in chemoresistant human ovarian cancer cells. AB - Cisplatin-centered chemotherapy is a key treatment for ovarian cancer, but resistance to chemotherapeutic agents remains a major cause of treatment failure. Multiple factors are known to contribute to the development of this chemoresistance. Although it has been demonstrated that X-linked inhibitor of apoptosis protein (Xiap) prevents apoptosis by inhibiting effector caspases, if and how it is important in chemoresistance in ovarian cancer has not been studied. The effects of Xiap down-regulation and/or restoration of wild type p53 by recombinant adenovirus infection were examined on four ovarian epithelial cancer cell lines [C13*, A2780-s (wild type p53), A2780-cp (mutant p53), and SKOV3 (null p53)]. Apoptosis and protein expression (e.g., Xiap, caspase-3, p53, MDM2, and p21waf1) were assessed by Hoechst 33258 stain and Western blot, respectively. We demonstrated that Xiap down-regulation following adenoviral antisense expression induces apoptosis in the wild-type p53 cells, but not in the mutated or null cells. Xiap down-regulation resulted in caspase-3 activation, caspase-mediated MDM2 processing, and p53 accumulation. Restoration of wild type p53 in the p53-mutated or -null cells significantly enhanced the proapoptotic effect of Xiap antisense expression. Down-regulation of Xiap induced apoptosis in chemoresistant ovarian cancer cells, a process dependent on p53 status. PMID- 11059758 TI - Histone H1 and H3 dephosphorylation are differentially regulated by radiation induced signal transduction pathways. AB - We recently demonstrated that linker histone H1, which is thought to have a fundamental role in higher-order chromatin structure, becomes transiently dephosphorylated after ionizing radiation (IR) in a mutated ataxia telangiectasia (ATM) dependent manner. To establish whether H1 dephosphorylation was a component of a damage-response pathway that included dephosphorylation of other histones, we asked whether H3 was dephosphorylated in response to IR in a manner similar to H1. H1 and H3 are maximally phosphorylated in metaphase and both are dephosphorylated after IR. However, the duration of IR-induced H3 dephosphorylation is significantly longer than that of IR-induced H1 dephosphorylation. Moreover, H1 dephosphorylation is ATM-dependent, whereas H3 dephosphorylation is ATM-independent. These observations suggest that the damage sensing pathways regulating H3 and H1 dephosphorylation diverge upstream of ATM. PMID- 11059759 TI - Interferon-gamma treatment elevates caspase-8 expression and sensitizes human breast tumor cells to a death receptor-induced mitochondria-operated apoptotic program. AB - In this report, we have assessed the role of IFN-gamma as a sensitizing agent in apoptosis mediated by activation of death receptor CD95 in breast tumor cells. Treatment of the tumor cell lines MCF-7 and MDA-MB231 with IFN-gamma significantly facilitated apoptosis induced by CD95 receptor ligation at the plasma membrane, independently of p53 status. In contrast, IFN-gamma treatment did not enhance the apoptotic effect of the DNA-damaging drug, doxorubicin. Analysis of apoptosis regulators indicated that caspase-8 mRNA and protein levels were up-regulated in both of the cell lines after treatment with IFN-gamma. Furthermore, IFN-gamma sensitized MCF-7 and MDA-MB231 cells to CD95-mediated activation of caspase-8, induction of cytochrome c release from mitochondria, and processing of caspase-9. Release of cytochrome c, caspases activation, and apoptosis were prevented in MCF-7 cells overexpressing Bcl-2. Altogether these results indicate that IFN-gamma, maybe through the elevation of caspase-8 levels, sensitizes human breast tumor cells to a death receptor-mediated, mitochondria operated pathway of apoptosis. PMID- 11059760 TI - The relationship between benzo[a]pyrene-induced mutagenesis and carcinogenesis in repair-deficient Cockayne syndrome group B mice. AB - Cockayne syndrome (CS) patients are deficient in the transcription coupled repair (TCR) subpathway of nucleotide excision repair (NER) but in contrast to xeroderma pigmentosum patients, who have a defect in the global genome repair subpathway of NER, CS patients do not have an elevated cancer incidence. To determine to what extent a TCR deficiency affects carcinogen-induced mutagenesis and carcinogenesis, CS group B correcting gene (CSB)-deficient mice were treated with the genotoxic carcinogen benzo(a)pyrene (B[a]P) at an oral dose of 13 mg/kg body weight, three times a week. At different time points, mutant frequencies at the inactive lacZ gene (in spleen, liver, and lung) as well as at the active hypoxanthine phosphoribosyltransferase (Hprt) gene (in spleen) were determined to compare mutagenesis at inactive versus active genes. B[a]P treatment gave rise to increased mutant frequencies at lacZ in all of the organs tested without a significant difference between CSB-/- and wild-type mice, whereas B[a]P-induced Hprt mutant frequencies in splenic T-lymphocytes were significantly more enhanced in CSB-/- mice than in control mice. The sequence data obtained from Hprt mutants indicate that B[a]P adducts at guanine residues were preferentially removed from the transcribed strand of the Hprt gene in control mice but not in CSB-/- mice. On oral treatment with B[a]P, the tumor incidence increased in both wild-type and CSB-deficient animals. However, no differences in tumor rate were observed between TCR-deficient CSB-/- mice and wild-type mice, which is in line with the normal cancer susceptibility of CS patients. The mutagenic response at lacZ, in contrast to Hprt, correlated well with the cancer incidence in CSB-/- mice after B[a]P treatment, which suggests that mutations in the bulk of the DNA (inactive genes) are a better predictive marker for carcinogen-induced tumorigenesis than mutations in genes that are actively transcribed. Thus, the global genome repair pathway of NER appears to play an important role in the prevention of cancer. PMID- 11059761 TI - Both (+/-)syn- and (+/-)anti-7,12-dimethylbenz[a]anthracene-3,4-diol-1,2-epoxides initiate tumors in mouse skin that possess -CAA- to -CTA- mutations at Codon 61 of c-H-ras. AB - We have determined the tumor-initiating activity of (+/-)syn- and (+/-)anti-7,12 dimethylbenz[a]anthracene-3,4-diol-1,2-epoxide (syn- and anti-DMBADE), the two metabolically formed bay-region diol epoxides of DMBA, and we have also analyzed mutations in the H-ras gene from tumors induced by these compounds. Using a two stage, initiation-promotion protocol for tumorigenesis in mouse skin, we have found that both syn- and anti-DMBADE are active tumor initiators, and that the occurrence of papillomas is carcinogen dose dependent. All of the papillomas induced by syn-DMBADE (a total of 40 mice), 96% of those induced by anti-DMBADE (a total of 25 mice), and 94% of those induced by DMBA (a total of 16 mice) possessed a -CAA- to -CTA- mutation at codon 61 of H-ras. No mutations in codons 12 or 13 were detected in any tumor. Topical application of syn- and anti-DMBADE produced stable adducts in mouse epidermal DNA, most of which comigrated with stable DNA adducts formed after topical application of DMBA. Further analysis of the data showed that levels of the major syn- and anti-DMBADE-deoxyadenosine adducts formed after topical application of DMBA are sufficient to account for the tumor-initiating activity of this carcinogen on mouse skin. Previously, we showed that both the syn- and anti-DMBADE bind to the adenine (A182) at codon 61 of H-ras. Collectively, these results indicate that the adenine adducts induced by both bay-region diol epoxides of DMBA lead to the mutation at codon 61 of H ras and, consequently, initiate tumorigenesis in mouse skin. PMID- 11059762 TI - Inhibition of ornithine decarboxylase (ODC) decreases tumor vascularization and reverses spontaneous tumors in ODC/Ras transgenic mice. AB - We have shown that ornithine decarboxylase (ODC) overexpression in the skin of TG.AC v-Ha-ras transgenic mice induces the formation of spontaneous skin carcinomas. Treatment of ODC/Ras double transgenic mice with alpha difluoromethylornithine (DFMO), a specific inhibitor of ODC enzyme activity, causes a rapid regression of these spontaneous tumors. DFMO treatment led to dramatic decreases in ODC activity and putrescine levels, but v-Ha-ras expression was not affected in the regressed tumors. Moreover, cyclin D1 continued to be strongly expressed in the basal epithelial cells of regressed tumors, and there was no decrease in the proliferative index of these same tumor cells. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling analyses revealed increased DNA fragmentation in DFMO regressed tumors compared with similarly sized spontaneous tumors from ODC/Ras transgenic mice not treated with DFMO. Moreover, the blood vessel count was significantly decreased in regressed tumors within the first four days of DFMO treatment. The decreased vasculature in DFMO regressed tumors was not attributable to altered expression of murine vascular endothelial growth factor (VEGF) isoforms. Elevated levels of ODC activity in the skin of K6/ODC transgenic mice increased the dermal vascularization compared with that in nontransgenic normal littermates. Our results suggest that ODC stimulates an angiogenic factor(s) other than VEGF and/or may play a key role in a cell survival effector pathway of Ras that is independent of a Ras-induced proliferation pathway. PMID- 11059763 TI - Estrogenic and antiproliferative properties of glabridin from licorice in human breast cancer cells. AB - There is an increasing demand for natural compounds that improve women's health by mimicking the critical benefits of estrogen to the bones and the cardiovascular system but avoiding its deleterious effects on the breast and uterus. The estrogenic properties of glabridin, the major isoflavan in licorice root, were tested in view of the resemblance of its structure and lipophilicity to those of estradiol. The results indicate that glabridin is a phytoestrogen, binding to the human estrogen receptor and stimulating creatine kinase activity in rat uterus, epiphyseal cartilage, diaphyseal bone, aorta, and left ventricle of the heart. The stimulatory effects of 2.5-25 microg/animal glabridin were similar to those of 5 microg/animal estradiol. Chemical modification of glabridin showed that the position of the hydroxyl groups has a significant role in binding to the human estrogen receptor and in proliferation-inducing activity. Glabridin was found to be three to four times more active than 2'-O-methylglabridin and 4' O-methylglabridin, and both derivatives were more active than 2',4'-O methylglabridin. The effect of increasing concentrations of glabridin on the growth of breast tumor cells was biphasic. Glabridin showed an estrogen receptor dependent, growth-promoting effect at low concentrations (10 nM-10 microM) and estrogen receptor-independent antiproliferative activity at concentrations of > 15 microM. This is the first study to indicate that isoflavans have estrogen-like activities. Glabridin and its derivatives exhibited varying degrees of estrogen receptor agonism in different tests and demonstrated growth-inhibitory actions on breast cancer cells. PMID- 11059764 TI - Increased risk of prostate cancer and benign prostatic hyperplasia associated with a CYP17 gene polymorphism with a gene dosage effect. AB - The CYP17 gene (CYP17) codes for the cytochrome P450c17alpha enzyme, which mediates two key steps in the sex steroid synthesis. There is a polymorphism (a T to-C substitution) in the 5'-untranslated region, which may influence the transcription level of CYP17 mRNA. There is a continuing controversy as to whether the variant allele is associated with a subset of breast cancer or polycystic ovary syndrome. In prostate cancer research, there are contradictory data concerning the CYP17 risk allele. We explored the association between CYP17 polymorphism and a risk of prostate cancer or benign prostatic hyperplasia (BPH) in a Japanese population. This study included 252 prostate cancer patients, 202 BPH patients, and 131 male controls. A 451-bp fragment encompassing the polymorphic site was amplified by PCR, treated with restriction enzyme MspA1, and electrophoresed on an agarose gel. The MspA1-undigested allele with the published sequence and the MspA1-digested variant allele were designated as A1 and A2, respectively. There was a significant difference (P < 0.05) in the genotypes between prostate cancer patients and male controls, and between BPH patients and male controls. Men with the A1/A1 CYP17 genotype had an increased risk of prostate cancer [odds ratio (OR), 2.57; 95% confidence interval (CI) = 1.39-4.78] and BPH (OR, 2.44; 95% CI = 1.26-4.72) compared with those with the A2/A2 genotype. Men with the A1/A2 genotype had an intermediate increased risk of prostate cancer (OR, 1.45; 95% CI = 0.84-2.54) and BPH (OR, 1.60; 95% CI = 0.89 2.87) compared with those with the A2/A2 genotype. The trend of an increasing risk of prostate cancer and BPH with an increasing number of the A1 allele was statistically significant (prostate cancer versus male control, P = 0.003; OR, 1.57; 95% CI = 1.16-2.12; BPH versus male control, P = 0.008; OR, 1.55; 95% CI = 1.12-2.13). There was no significant association between the CYP17 genotype and the tumor status (grade and stage) of prostate cancer. Our results suggest that the A1 allele of the CYP17 polymorphism is associated with an increased risk of prostate cancer and BPH, with a gene dosage effect. However, the CYP17 genotype does not seem to influence the disease status in prostate cancer. PMID- 11059765 TI - Role of the immune response during neuro-attenuated herpes simplex virus-mediated tumor destruction in a murine intracranial melanoma model. AB - Neuro-attenuated herpes simplex virus-1 (HSV-1) gamma34.5 mutants can slow progression of preformed tumors and lead to complete regression of some tumors. However, the role of the immune response in this process is poorly understood. Syngenic DBA/2 tumor-bearing mice treated with HSV-1 1716 fourteen days after tumor implantation had significant prolongation in survival when compared with mice treated with viral growth sera (mock; 38.9 +/- 2.3 versus 24.9 +/- 0.6, respectively; P < 0.0001). Additionally, 60% of the animals treated on day 7 had complete regression of the tumors. However, no difference was observed in the mean survival rates of viral- or mock-treated tumor-bearing SCID mice (15 +/- 1.7 versus 14.8 +/- 2.2, respectively). When DBA/2 mice syngenic for the tumor were depleted of leukocytes by cyclophosphamide administration (before and during viral administration), there was again no significant difference observed in the survival times (19.0 +/- 1.9 versus 19.5 +/- 2.7, respectively). These data demonstrate that the immune response contributes to the viral-mediated tumor destruction and the increase in survival. Immune cell infiltration was up regulated, specifically CD4+ T cells and macrophages (which are found early after viral administration). Prior immunity to HSV-1 increased survival times of treated mice over those of naive mice, an important consideration because 50-95% of the adult human population is sero-positive for HSV-1. Our results imply that the timing of viral administration and the immune status of the animals will be an important consideration in determining the effectiveness of viral therapies. PMID- 11059766 TI - Treatment of colorectal liver metastases by adenoviral transfer of tissue inhibitor of metalloproteinases-2 into the liver tissue. AB - Metastatic disease is the leading cause of death in cancer patients. Here, we describe a novel gene therapeutic strategy for prevention of metastatic spread by providing a suitable defense mechanism for the target organ. The production of metalloproteinase (MMP) enzymes by cancer cells is critical for local invasion and for infiltration of metastatic cells into distant sites. Using a nude mouse model of colorectal liver metastasis, we have overexpressed the MMP inhibitor, tissue inhibitor of MMP-2 (TIMP-2) in the liver prior to, or following, tumor challenge by metastatic LS174T cells in vivo. Transduction of approximately 50% of hepatocytes resulted in 95% reduction in metastasis after tumor challenge compared with controls. Furthermore, TIMP-2 gene transfer into livers with preexisting metastatic spread resulted in a 77% reduction in tumor cell growth. Our data imply that MMP activity of metastatic cancer cells is required for spread and subsequent tumor growth and that enhancing antiproteolytic defense mechanisms in target organs represents a novel form of cancer gene therapy. PMID- 11059767 TI - The human leukemic T-cell line, TALL-104, is cytotoxic to human malignant brain tumors and traffics through brain tissue: implications for local adoptive immunotherapy. AB - Preclinical studies with the human MHC nonrestricted cytotoxic T-cell leukemic line, TALL-104, were performed in anticipation of its use in cellular immunotherapy trials for primary malignant brain tumors. In this study, we have: (a) quantitated the in vitro brain tumor cell lysis; (b) measured the cytokine secretion upon coincubation of TALL-104 cells with brain tumor cells; (c) investigated the effect of dexamethasone on brain tumor cell cytolysis by TALL 104 cells; (d) explored the effects of lethal irradiation and cryopreservation on TALL-104 cell viability and lytic efficacy; and (e) estimated the damage TALL-104 cells induce to murine normal and tumor brain cells and their trafficking patterns in both normal and tumor-bearing rat brain upon intracranial infusion. In vitro coincubation of TALL-104 cells with human brain tumor cells, explants, and cell lines resulted in significant lysis of them, but normal brain cells were spared. Lysis of tumor at 4 h was unaffected by dexamethasone or lethal irradiation. Secretion of tumor necrosis factor-alpha, tumor necrosis factor beta, IFN-gamma, or granulocyte/macrophage-colony stimulating factor upon TALL 104 cell coincubation with brain tumor cells variably occurred without always correlating with lysis. In vivo experiments using irradiated TALL-104 cells, placed at multiple times into normal cannulated rat brain, produced focal sterile abscesses at the instillation site but no widespread allergic encephalitic reaction. Cells morphologically consistent with TALL-104 cells specifically trafficked from the site of instillation through the neuropil, occasionally into the contralateral brain, and egressed at perivascular and leptomeningeal spaces. In vivo experiments with cannulated rats bearing 9L gliosarcoma showed a preferential localization of the TALL-104 cells in tumor compared with normal brain. Taken together, these data support the concept that TALL-104 cells can be used as a novel nontoxic and efficacious paradigm for cellular immunotherapy trials in human primary malignant brain tumors. PMID- 11059768 TI - CM101 treatment overrides tumor-induced immunoprivilege leading to apoptosis. AB - CM101, a bacterial polysaccharide exotoxin produced by group B Streptococcus (GBS), also referred to as GBS toxin, has been shown to target pathological neovasculature and activate complement (C3), thereby inducing neovascularitis, infiltration of inflammatory cells, inhibition of tumor growth, and apoptosis in murine tumor models. Data from refractory cancer patients in a Phase I clinical trial with CM101 indicated a similar mechanism of tumor-targeted inflammation. To further our understanding of the mechanism of action of CM101 as an antitumor agent, we examined the role of the inflammatory response in inducing tumor apoptosis in a normal mouse and tumor-bearing mouse model. The i.v. infusion of CM101 into B16BL-6 melanoma tumor-bearing mice elevated p53 mRNA in circulating leukocytes as measured by reverse transcription-PCR, and immunohistochemistry demonstrated infiltration and sequestration of leukocytes. Whole tumor lysates from excised tumors exhibited an increase in binding to the murine p21(Waf1/Cip1) derived p53 DNA binding sequence compared with control whole tumor lysates, in which minimal or no DNA binding was observed. CM101 infusion led to elevated levels of Fas protein within the tumors as well as a decrease in the expression of fas ligand (fasL). Furthermore, tumors were apoptotic as determined by terminal deoxynucleotidyl transferase-mediated nick end labeling and DNA fragmentation assays. Collectively, these data suggest that CM101 up-regulates p53 in tumor-infiltrating leukocytes, initiating a loss of tumor immunoprivilege and consequently rendering the tumor sensitive to Fas/fasL-mediated apoptosis. CM101 induced loss of tumor immunoprivilege through changes in the expression of leukocyte p53, tumor Fas and fasL coupled with neovascularitis and leukocyte infiltration, constitutes a plausible molecular pathway for tumor reduction observed in cancer patients. PMID- 11059769 TI - Changes in subcellular distribution of topoisomerase IIalpha correlate with etoposide resistance in multicell spheroids and xenograft tumors. AB - The outer cells of Chinese hamster V79 spheroids are about 10 times more resistant than monolayers to DNA damage and cell killing by the topoisomerase (topo) II inhibitor etoposide. Although the amount and catalytic activity of topo IIalpha are identical for monolayers or the outer cells of spheroids, and the cell proliferation rate is the same, our previous results indicated that phosphorylation of topo IIalpha is at least 10 times higher in V79 monolayers than in spheroids. Because phosphorylation of topo IIalpha has been associated with nuclear translocation, we examined subcellular distribution of Topo IIalpha in monolayers, spheroids, and xenograft tumors using immunohistochemistry. Topo IIalpha was located predominantly in the nucleus of V79, human SiHa, and rat C6 monolayers but was found mainly in the cytoplasm of the proliferating outer cells of spheroids formed from these cell lines. Conversely, the outer cells of WiDr human colon carcinoma spheroids showed predominantly nuclear localization of topo IIalpha, and only WiDr cells showed no increase in resistance to etoposide when grown as spheroids. Cells sorted from xenografts resembled the spheroids in terms of sensitivity to etoposide and location of topo IIalpha. When the outer cells of V79 spheroids were returned to monolayer growth, the rate of redistribution of topo IIalpha to the nucleus occurred with similar kinetics as the increase in sensitivity to killing by etoposide. Removal and return of individual outer V79 spheroid cells to suspension culture resulted in the translocation of topo IIalpha to the nucleus for the first 24 h, accompanied by an increase in sensitivity to DNA damage by etoposide. Therefore, the cytoplasmic topo IIalpha distribution in outer spheroid cells and tumors appears to correlate not with morphological changes associated with growth in suspension but rather with the presence of neighboring, noncycling cells. PMID- 11059770 TI - Apo2 ligand/TNF-related apoptosis-inducing ligand and death receptor 5 mediate the apoptotic signaling induced by ionizing radiation in leukemic cells. AB - Ionizing radiation is a major tool for cancer treatment. The response of eukaryotic cells to ionizing radiation includes apoptosis, a process which requires activation of multiple genes. We sought to determine whether radiation induced gene expression plays a role in radiation-induced apoptosis. We found Apo2 ligand (Apo2L, also called TRAIL) mRNA induction following gamma-irradiation of Jurkat, MOLT-4, CEM, and PBMC, all human T lineage-derived cells. Increased Apo2L protein levels were found in MOLT-4 and Jurkat cells. Radiation also activated the Apo2L death receptor (DR)5 (also called Apo2, TRAIL-R2, or KILLER) in MOLT-4 cells, which harbor a wild-type p53. We isolated 1152 bp of 5' flanking region of the Apo2L gene and a shorter fragment of 716 bp, both of which showed promoter activity driving the expression of a luciferase reporter gene; however, the response to radiation in MOLT-4 cells was lost when only 430 bp of 5' proximal flanking sequence was maintained. Exogenous Apo2L induced phosphatidylserine exposure on cell membranes, caspase 8 and caspase 3 activation, key markers of apoptosis, confirming that the Apo2L/DR5 pathway is functional in these cells. Bid, a Bcl-2 family protein also known to contribute to receptor-mediated apoptosis, was also activated. To determine whether Apo2L and DR5 were critical for radiation signaling to apoptosis, we stably expressed a dominant negative DR5delta-receptor in Jurkat cells. Cell survival was significantly augmented, indicating that increased Apo2L expression contributed to radiation-induced apoptosis. Clonogenic assays demonstrated that purified, recombinant soluble Apo2L enhanced the lethality of low, therapeutic doses (1-2 Gy) of gamma-irradiation. These data suggest that production of Apo2L may cooperate synergistically with the cytotoxic effect of radiation, and that combinations of Apo2L and radiation may become a powerful tool in clinical therapy. PMID- 11059771 TI - Extensive contribution of the multidrug transporters P-glycoprotein and Mrp1 to basal drug resistance. AB - Despite accumulating evidence that multidrug resistance transporter proteins play a part in drug resistance in some clinical cancers, it remains unclear whether the relatively low levels of multidrug resistance transporter expression found in most untreated tumors could substantially affect their basal sensitivity to antineoplastic drugs. To shed light on this problem, the drug sensitivities of wild-type mouse cell lines were compared with those of lines in which the Mdr1a and Mdr1b genes encoding P-glycoprotein (P-gp) were inactivated and lines in which the Mrp1 gene was inactivated in addition to Mdr1a and Mdr1b. These models permit a clean dissection of the contribution of each transporter to drug resistance at expression levels similar to those in normal tissues and avoid complications that might arise from previous exposure of cell lines to drug selection. For substrate drugs, we found that these contributions can indeed be very substantial. Lines lacking functional P-gp were, on average, markedly more sensitive to paclitaxel (16-fold), anthracyclines (4-fold) and Vinca alkaloids (3 fold). Lines lacking both P-gp and Mrp1 were (compared with wild-type lines) hypersensitive to an even broader array of drugs, including epipodophyllotoxins (4-7-fold), anthracyclines (6-7-fold), camptothecins (3-fold), arsenite (4-fold) and Vinca alkaloids, especially vincristine (28-fold). Thus, even very low levels of P-gp and Mrp1 expression that may be difficult to detect in tumors could significantly affect their innate sensitivity to a wide range of clinically important substrate drugs. An implication is that the use of resistance reversal agents to sensitize drug-naive tumors may be appropriate in more cases than is presently appreciated. PMID- 11059772 TI - Selective inhibition of cyclooxygenase-2 suppresses growth and induces apoptosis in human esophageal adenocarcinoma cells. AB - Adenocarcinoma in Barrett's esophagus has been increasing in incidence at a rapid rate for more than two decades. Cyclooxygenase (COX)-2 appears to play an important role in gastrointestinal carcinogenesis, and COX-2 overexpression has been demonstrated both in esophageal adenocarcinomas and in the metaplastic epithelium of Barrett's esophagus. The aim of our study was to determine whether selective inhibition of COX-2 by NS-398 would alter the rates of cell growth and apoptosis in human Barrett's-associated esophageal adenocarcinoma cell lines. COX 1 and COX-2 expression in adenocarcinoma cell lines was determined using reverse transcription-PCR and Western blotting for mRNA and protein, respectively. Esophageal adenocarcinoma cell lines were treated with various concentrations of NS-398 (selective for COX-2 inhibition) and flurbiprofen (selective for COX-1 inhibition). Cell growth was compared in flurbiprofen-treated and untreated tumor cell lines; cell growth and apoptosis were compared in NS-398-treated and untreated tumor cell lines. COX-2 mRNA and protein were detected in two of three cell lines (SEG-1 and FLO); the third cell line, BIC-1, did not express COX-2 mRNA or protein under basal conditions or after stimulation with phorbol 12 myristate 13-acetate. Treatment with COX-1-selective concentrations of flurbiprofen did not affect cell growth in any of the three tumor cell lines. In contrast, treatment with COX-2-selective concentrations of NS-398 significantly suppressed cell growth and increased apoptosis in the cell lines that expressed COX-2 (SEG-1 and FLO), but not in the cell line that did not express COX-2 (BIC 1). We conclude that the administration of a selective inhibitor of COX-2 significantly decreases cell growth and increases apoptosis in Barrett's associated adenocarcinoma tumor cells that express COX-2. These observations suggest a potential role for selective COX-2 inhibitors in the prevention and treatment of esophageal adenocarcinoma for patients with Barrett's esophagus. PMID- 11059773 TI - Selective radiosensitization of drug-resistant MutS homologue-2 (MSH2) mismatch repair-deficient cells by halogenated thymidine (dThd) analogues: Msh2 mediates dThd analogue DNA levels and the differential cytotoxicity and cell cycle effects of the dThd analogues and 6-thioguanine. AB - Mismatch repair (MMR) deficiency, which underlies hereditary nonpolyposis colorectal cancer, has recently been linked to a number of sporadic human cancers as well. Deficiency in this repair process renders cells resistant to many clinically active chemotherapy agents. As a result, it is of relevance to find an agent that selectively targets MMR-deficient cells. We have recently shown that the halogenated thymidine (dThd) analogues iododeoxyuridine (IdUrd) and bromodeoxyuridine (BrdUrd) selectively target MutL homologue-1 (MLH1)-deficient human cancer cells for radiosensitization. The levels of IdUrd and BrdUrd in cellular DNA directly correlate with the ability of these analogues to increase the sensitivity of cells and tissues to ionizing radiation, and data from our laboratory have demonstrated that MLH1-mediated MMR status impacts dThd analogue DNA levels, and consequently, analogue-induced radiosensitization. Here, we have extended these studies and show that, both in human and murine cells, MutS homologue-2 (MSH2) is also involved in processing dThd analogues in DNA. Using both E1A-transformed Msh2+/+ and Msh2-/- murine embryonic stem (ES)-derived cells (throughout this report we use Msh2+/+ and Msh2-/- to refer to murine ES-derived cell lines that are wild type or mutant, respectively, for the murine Msh2 gene) and human endometrial cancer cells differing in MSH2 status, we see the classic cytotoxic response to 6-thioguanine (6-TG) in Msh2+/+ and human HEC59/2-4 (MSH2+) MMR-proficient cells, whereas Msh2-/- cells and human HEC59 (MSH2-/-) cells are tolerant (2-log difference) to this agent. In contrast, there is very little cytotoxicity in Msh2+/+ ES-derived and HEC59/2-4 cells to IdUrd, whereas Msh2-/- and HEC59 cells are more sensitive to IdUrd. High-performance liquid chromatography analysis of IdUrd and BrdUrd levels in DNA suggests that this differential cytotoxicity may be due to lower analogue levels in MSH2+ murine and human tumor cells. The DNA levels of IdUrd and BrdUrd continue to decrease over time in Msh2+/+ cells following incubation in drug-free medium, whereas they remain high in Msh2-/- cells. This trend was also found in MSH2-deficient human endometrial cancer cells (HEC59) when compared with HEC59/2-4 (hMsh2-corrected) cells. As a result of higher analogue levels in DNA, Msh2-/- cells are selectively targeted for radiosensitization by IdUrd. Fluorescence-activated cell sorting analysis of Msh2+/+ and Msh2-/- cells shows that selective toxicity of the halogenated nucleotide analogues is not correlated with a G2-M cell cycle arrest and apoptosis, as is found for selective killing of Msh2+/+ cells by 6-TG. Together, these data demonstrate MSH2 involvement in the processing of IdUrd and BrdUrd in DNA, as well as the differential cytotoxicity and cell cycle effects of the halogenated dThd analogues compared with 6-TG. Therefore, IdUrd and BrdUrd may be used clinically to selectively target both MLH1- and MSH2-deficient, drug resistant cells for radiosensitization. PMID- 11059774 TI - Activation of lytic Epstein-Barr virus (EBV) infection by radiation and sodium butyrate in vitro and in vivo: a potential method for treating EBV-positive malignancies. AB - The consistent presence of the EBV genome in certain tumors offers the potential for novel EBV-directed therapies. Switching the latent form of EBV infection present in most EBV-positive tumor cells into the cytolytic form may be clinically useful because lytic EBV infection leads to host cell destruction, and very few normal cells contain the EBV genome. It would also be therapeutically advantageous to induce expression of EBV-encoded lytic proteins that convert the nucleoside analogues ganciclovir (GCV) and 3'-azido-3'deoxythymidine (AZT) into their active, cytotoxic forms. In this report, we have explored two different approaches for activating the lytic form of EBV infection in tumors. We show that gamma-irradiation at clinically relevant doses induces lytic EBV infection in lymphoblastoid cell lines in vitro as well as in EBV-positive B-cell tumors in SCID mice. In addition, sodium butyrate (given as a single i.p. dose) is effective for activating lytic viral infection in some EBV tumor types in SCID mice. We also examined whether low-dose gamma-irradiation treatment of EBV positive lymphoblastoid cells in vitro promotes GCV or AZT susceptibility. The combination of radiation with either GCV or AZT induced significantly more cell killing in vitro than either radiation or prodrug treatment alone. Most importantly, we found that the combination of gamma-irradiation and GCV was much more effective in treating EBV-positive lymphoblastoid tumors in SCID mice than either agent alone. Thus, GCV or AZT treatment could potentially enhance the therapeutic efficacy of radiation therapy for EBV-positive lymphomas in patients. PMID- 11059775 TI - Loss of interferon-gamma inducibility of TAP1 and LMP2 in a renal cell carcinoma cell line. AB - The inadequate ability of cancer cells to present antigen on the cell surface via MHC class I molecules is one mechanism by which tumor cells evade antitumor associated antigen immunity. In many cases, such as in renal cell carcinoma (RCC), the lack of MHC class I antigen presentation can be attributed to the down regulation of genes needed for antigen processing, such as the transporters associated with antigen processing (TAP)1 and TAP2, and the proteasomal components low molecular weight proteins (LMP)2 and LMP7. The TAP1 and LMP2 genes are transcribed from a shared bidirectional promoter containing an IFN response factor element that confers IFN-gamma inducibility. Here, we investigate the differential responsiveness to IFN-gamma of RCC cell lines, Caki-1 and Caki-2, which have been reported to have abnormally low expressions of TAP1 and LMP2. We now demonstrate that the Caki-2 cell line is defective in the IFN-gamma signaling pathway. The effects of IFN-gamma on TAP1 and LMP2 expression revealed a loss of up-regulation in Caki-2 cells, but not in Caki-1 cells. In vivo DNA footprinting shows a specific loss of occupancy at the IFN response factor element site in Caki-2 cells, whereas Caki-1 cells show full promoter occupancy. Furthermore, in vitro DNA-binding studies indicated that Caki-2 cells do not have IFN-regulatory factor 1- or signal transducer and activator of transcription 1 (Stat1)-binding activity after IFN-gamma stimulation. Examination of Stat1, Jak1, and Jak2 proteins demonstrated that the proteins were expressed, however, not phosphorylated, upon IFN-gamma treatment in Caki-2 cells. Also, this cell line expressed both IFN-gamma receptor chains. IFN-gamma inducibility could not be rescued by introduction of normal Jak1 and/or Jak2 proteins. However, overexpression of Jak1 did increase TAP1 and LMP2 expression independent of IFN gamma, indicating that the Stat1 and IFN-regulatory factor 1 proteins present in Caki-2 can be activated. These findings suggest that the loss of TAP1 and LMP2 induction is a defect in the earliest steps of the IFN-gamma signaling pathway resulting in the inability of Caki-2 cells to up-regulate the MHC class I antigen processing pathway. Because immunotherapy may be one of the most promising approaches for treating RCC, understanding the mechanisms by which these tumors circumvent cytokine signaling, thereby evading antitumor-specific-antigen immunity, would greatly aid the efficacy of such therapy. PMID- 11059776 TI - Pretransplant tumor antigen-specific immunization of allogeneic bone marrow transplant donors enhances graft-versus-tumor activity without exacerbation of graft-versus-host disease. AB - Allogeneic bone marrow transplantation (BMT) causes a beneficial graft-versus tumor (GVT) immune response that is often associated with graft-versus-host disease (GVHD). There is substantial interest in developing therapeutic strategies that augment GVT without GVHD. We have demonstrated recently that immunization of BMT donors with cellular tumor vaccines leads to curative GVT but induces unacceptable GVHD because of the presence of recipient minor histocompatibility antigens (mHAgs) in whole-cell tumor vaccines. This study tested the hypothesis that immunization of BMT donors against a defined tumor specific antigen with a vaccine not containing recipient mHAgs would help to separate the two responses by enhancing GVT activity without exacerbating GVHD, even when cellular vaccines were used after BMT. Recipient strain C57BL/6 fibrosarcoma cells engineered to express the well-characterized model tumor antigen, influenza nucleoprotein (NP), were used in these studies. C3H.SW donors were immunized against NP prior to BMT, and cytolytic T cells were transferred along with bone marrow into irradiated H-2-matched, mHAg-mismatched C57BL/6 recipients with established micrometastatic 205-NP tumors. Donor immunization led to a significant increase in GVT activity, as measured by reduction in tumor growth and enhanced survival. However, deaths in recipients of tumor antigen specific immune BMT ultimately occurred because of the growth of antigen-loss variants; such tumor growth did not occur in animals receiving BMT from donors treated with whole-cell vaccines. Donor immunization did not lead to an exacerbation of GVHD, even when BMT recipients received additional immunization after BMT with a 205-NP "whole" tumor cell vaccine (which was shown to induce fatal GVHD when used for donor immunization). In conclusion, immunization of allogeneic BMT donors against a tumor-specific antigen significantly enhances GVT activity without an associated exacerbation of GVHD. PMID- 11059777 TI - Hormone-dependent tumor regression in vivo by an inducible transcriptional repressor directed at the PAX3-FKHR oncogene. AB - In alveolar rhabdomyosarcomas (ARMSs), a specific chromosomal translocation creates a fusion transcription factor, PAX3-FKHR, that is oncogenic due to transcriptional activation. As a strategy for down-regulation of PAX3-FKHR target genes, we created conditional PAX3 repressors by fusing the PAX3 DNA-binding motifs to the hormone binding domain (HBD) of the estrogen receptor and to the KRAB repression domain. We validated proper expression, specific DNA binding, corepressor interaction, and nuclear localization for the KRAB-PAX3-HBD protein and showed it to be a 4-hydroxytamoxifen-dependent transcriptional repressor of transiently transfected and integrated PAX3 reporters in ARMS cells. We established ARMS cell lines that exhibited stable expression of the conditional PAX3 repressor proteins and used them to down-regulate the malignant growth under low serum or anchorage-independent conditions in a hormone-dependent manner. Terminal deoxynucleotidyl transferase-mediated nick end labeling assays revealed that hormonal activation of the PAX3 repressors induced extensive apoptosis that correlated with down-regulation of BCL-X(L) expression. SCID mice that were engrafted with the KRAB-PAX3-HBD ARMS cell lines and were implanted with 4 hydroxytamoxifen timed-release pellets exhibited suppression of tumor growth and an altered vascularity that was not observed in the control mice. These observations strongly suggest that we have directly repressed the PAX3 target genes that are deregulated by the PAX3-FKHR oncogene in ARMS. PMID- 11059778 TI - Apoptosis induced by DNA damage O6-methylguanine is Bcl-2 and caspase-9/3 regulated and Fas/caspase-8 independent. AB - In the therapy of various kinds of tumors, methylating agents generating O6 methylguanine (O6MeG) in DNA are used. We studied the molecular mechanism of cell death induced by these agents by comparing isogenic cell lines proficient (MGMT+) and deficient (MGMT-) for the DNA repair protein alkyltransferase and exhibiting the tolerance phenotype. Hypersensitivity to methylation-induced cell killing of MGMT- cells is attributable to the potent induction of apoptosis. We show that apoptosis is a late event occurring >48 h after methylation. It was preceded by decrease in Bcl-2 protein level and accompanied by activation of caspase-9 and caspase-3. We also observed cytochrome c release and hypophosphorylation of Bad. Other members of the Bcl-2 family (Bag-1, Bak, Bax, and Bcl-xL) were not altered in expression. Transfection of MGMT- cells with bcl-2 protected against methylation-induced apoptosis, indicating that Bcl-2 plays a key role in the response. Induction of apoptosis in MGMT- cells was not triggered by Fas and Fas ligand (CD95, Apo-1) because both proteins remained unaltered in expression and receptor-proximal caspase-8 was not activated after methylation. Also, inhibition of caspase-8 was ineffective in modifying the apoptotic response, whereas inhibition of caspase-3 and caspase-9 blocked apoptosis. Tolerant cells that are unable to repair O6MeG and are impaired in mismatch repair were less sensitive regarding the induction of apoptosis and Bcl-2 decline, supporting the view that O6MeG-induced apoptosis requires mismatch repair. The ultimate O6MeG-derived lesions triggering the apoptotic pathway are likely to be DNA double-strand breaks, which were significantly formed in MGMT- but not in MGMT+ and tolerant cells and which preceded apoptosis. Overall, the data indicate that O6MeG induces apoptosis via secondary lesions that trigger Bcl-2 decline, cytochrome c release, and caspase-9 and caspase-3 activation independently of Fas/Fas ligand and p53, for which the cells are mutated. PMID- 11059779 TI - Ligand-independent activation of the androgen receptor by the differentiation agent butyrate in human prostate cancer cells. AB - Androgens are potent differentiation agents that regulate prostate-specific antigen (PSA) gene expression via the androgen receptor (AR) that binds to androgen response elements (AREs) on the PSA gene to initiate transcription. However, in the absence of androgens, PSA gene expression can become elevated. This suggests that either the AR can be activated in the absence of androgen to elevate PSA gene expression through AREs on the PSA gene or that another transcription factor acting on the PSA promoter is stimulated. We have previously shown in vivo that butyrate, a differentiation agent that causes cell cycle arrest, increases serum PSA levels in castrated animals. Therefore, to determine the mechanism of butyrate induction of PSA, we used the LNCaP human prostate cancer cell line. Northern analyses and transfection experiments using a PSA reporter plasmid demonstrated induction of PSA gene expression by butyrate in LNCaP cells. Application of the antiandrogen bicalutamide blocked the induction of PSA mRNA by butyrate, suggesting a mechanism dependent on the AR. Consistent with this conclusion, electromobility shift assays showed increased AR-ARE complex formation with nuclear extracts from butyrate-treated cells. In addition, other reporter gene constructs that contain AREs were also induced by butyrate. Western blot analysis showed an increase in nuclear levels of AR protein in cells exposed to butyrate, whereas whole cell levels remained unchanged, suggesting that butyrate causes nuclear translocation of the AR. Thus, the differentiation agent butyrate causes ligand-independent activation of the AR to increase expression of the differentiation marker PSA in human prostate cancer cells. PMID- 11059780 TI - RhoC GTPase, a novel transforming oncogene for human mammary epithelial cells that partially recapitulates the inflammatory breast cancer phenotype. AB - Inflammatory breast cancer (IBC) is the most aggressive form of breast cancer and is phenotypically distinct from other forms of locally advanced breast cancer. In a previous study, we identified specific genetic alterations of IBC that could account for a highly invasive phenotype. RhoC GTPase was overexpressed in 90% of IBC archival tumor samples, but not in stage-matched, non-IBC tumors. To study the role of RhoC GTPase in contributing to an IBC-like phenotype, we generated stable transfectants of human mammary epithelial cells overexpressing the RhoC gene. The HME-RhoC transfectants formed large colonies under anchorage independent growth conditions, were more motile, and were invasive. In conjunction with an increase in motility, overexpression of RhoC led to an increase in actin stress fiber and focal adhesion contact formation. Furthermore, orthotopic injection into immunocompromised mice led to tumor formation. Taken together, these data indicate that RhoC GTPase is a transforming oncogene in human mammary epithelial cells and can lead to a highly invasive phenotype, akin to that seen in IBC. PMID- 11059781 TI - Tumor development is retarded in mice lacking the gene for urokinase-type plasminogen activator or its inhibitor, plasminogen activator inhibitor-1. AB - In vivo tumor progression in mice with targeted deficiencies in urokinase-type plasminogen activator (UPA-/-) and its inhibitor, plasminogen activator inhibitor 1 (PAI-1-/-), was studied using a fibrosarcoma tumor model. Murine T241 fibrosarcoma cells were s.c. implanted into three groups of mice with the following genotypes, wild-type (WT), UPA-/-, and PAI-1-/-. A significantly diminished primary tumor growth in UPA-/- and PAI-1-/- mice occurred, relative to WT mice. Tumors in UPA-/- and PAI-1-/- mice displayed lower proliferative and higher apoptotic indices and displayed a different neovascular morphology, as compared with WT mice. These results are consistent with the decreased growth rates of this tumor in these gene-deleted mice. Immunohistochemical analyses of the tumors revealed a decrease in vascularity and vascular endothelial growth factor expression only in tumors in PAI-1-/- mice. Analyses of the relative extents of corneal angiogenesis in these same animals, induced by basic fibroblast growth factor, corroborated the resistance of PAI-1-/- mice to neovascularization. The results obtained suggest that the host fibrinolytic system plays an important role in tumor growth in this model. Alterations in host expression of components of this system may alter tumor growth and dissemination by affecting the balance between tumor cell death and proliferation, as well as extracellular matrix changes needed for invasiveness and angiogenesis. PMID- 11059782 TI - A new Mr 55,000 surface protein implicated in melanoma progression: association with a metastatic phenotype. AB - Emergence of the invasive phenotype is a key event in the progression of human melanoma from benign proliferative lesions to malignant lesions. Recently we successfully selected in vivo from a poorly metastatic M4Beu. human melanoma cell line two variants (7GP and T1P26) that generate a higher frequency of spontaneous metastases to the lungs into immune-suppressed neonatal rats. Both cell lines showed no significant differences in the integrin profile of the subunits analyzed except for beta3, which was reduced to a background level in metastatic variants. To investigate how these variant sublines of human melanomas manage to sustain growth in the absence of alpha(v)beta3, a subtractive immunization approach was used to elicit host antibody response against cell surface proteins expressed on metastatic variants. In this study, a new monoclonal antibody (MoAb), LY1, that is highly specific for the 7GP and T1P26 variants, was isolated. LY1 identifies a membrane protein of Mr 55,000 on melanoma variants with epitopes that were resistant to sugar-cleaving enzymes. Immunostaining cells from variants by LY1 showed that staining is distributed to the cell periphery with high labeling intensity at the cell-to-cell contact points. This MoAb significantly inhibited invasion of metastatic variants through a reconstituted basement membrane (Matrigel) in vitro. Moreover, tumor growth of melanoma variants was dramatically affected in vivo with this MoAb. In vitro studies indicate that the LY1 MoAb does not inhibit chemotactic migration of the metastatic variants, the adhesion of tumor cells to vitronectin, collagen IV, fibronectin, and laminin, or cell proliferation. Expression of this antigen is high in human striated muscle, heart, spleen, brain, and lung and absent in kidney, liver, and pancreas. Using 59 fixed, paraffin-embedded archival tissues of human melanomas and nevi, LY1-reactive cells were not observed in melanocytes, nevi, or radial growth phase primary melanomas. In sharp contrast, LY1 selectively stained melanocytes derived from the vertical growth phase of many primary melanomas and metastatic melanomas. These results provide evidence that the Mr 55,000 protein expressed by selected variants with increased metastatic properties in vivo plays a functionally important role in determining metastasis. This molecule may represent a new metastatic risk marker in human melanoma and may be of biological importance in the identification of fatal metastatic subpopulations that have acquired competence for metastasis production. PMID- 11059783 TI - Reduced infiltration of tumor-associated macrophages in human prostate cancer: association with cancer progression. AB - Tumor-associated macrophages (TAMs) are highly active immune effector cells that may either positively or negatively regulate the growth of various malignant cells, depending on the biological context. However, the role of TAMs in human prostate cancer progression is unclear. TAMs were immunohistochemically labeled using a monoclonal (CD68) antibody in radical prostatectomy specimens derived from 81 prostate cancer patients. CD68-positive cells were counted with the aid of a microscope and expressed as macrophage index (MphiI), including TAMs/mm2 total tumor tissue (MphiItotal), TAMs/mm2 tumor stroma (MphiIstroma), and TAMs/mm2 cancer cell area (MphiIcancer). MphiIs were analyzed in association with patients' clinical and pathological stage, recurrence status, and histological grade of the cancer. There were significant inverse relationships between MphiItotal and MphiIstroma and clinical stage (P = 0.016 and P = 0.006, respectively). Reduced MphiItotal was also associated with the presence of positive lymph nodes (P = 0.010). Interestingly, although all of the MphiIs differed between Gleason score groups, only MphiIcancer was positively associated with Gleason score. Univariate analysis of MphiItotal and multivariate analysis of MphiItotal with specific pathological markers revealed that MphiItotal was an independent predictor for disease-free survival after surgery (Cox proportional hazard model, P = 0.044 and P = 0.007, respectively). For patients with high MphiItotal (> or = 185.8, the mean MphiItotal value), the disease-free probability 5 years after surgery was 0.75, which was significantly higher than for those with low MphiItotal (0.31, P = 0.0008). Additional immunohistochemical studies that evaluated cytotoxicity-related biomarkers in stroma-associated mononuclear cells suggested reduced functional activities in highly aggressive prostate cancer compared with less aggressive disease. Our results indicate that reduced MphiItotal is a novel prognostic marker for prostate cancer. PMID- 11059784 TI - B16 melanoma cell arrest in the mouse liver induces nitric oxide release and sinusoidal cytotoxicity: a natural hepatic defense against metastasis. AB - The formation of liver metastases involves interactions between intravascular cancer cells and the hepatic microvasculature. Here we provide evidence that the arrest of intravascular B16F1 melanoma cells in the liver induces a rapid local release of nitric oxide (NO) that causes apoptosis of the melanoma cells and inhibits their subsequent development into hepatic metastases. B16F1 melanoma cells (5 x 10(5)) labeled with fluorescent microspheres were injected into the portal circulation of C57BL/6 mice. The production of NO in vivo was detected by electron paramagnetic resonance spectroscopy ex vivo using an exogenous NO trapping agent. A burst of NO was observed in liver samples examined immediately after tumor cell injection. The relative electron paramagnetic resonance signal intensity was 667 +/- 143 units in mice injected with tumor cells versus 28 +/- 5 units after saline injection (P < 0.001). Two-thirds of cells arrested in the sinusoids compared with the terminal portal venules (TPVs). By double labeling of B16F1 cells with fluorescent microspheres and a TdT-mediated UTP end labeling assay, we determined that the melanoma cells underwent apoptosis from 4-24 h after arrest. The mean rate of apoptosis was 2-fold greater in the sinusoids than in the TPVs at 4, 8, and 24 h after injection (P < 0.05-0.01). Apoptotic cells accounted for 15.9 +/- 0.8% of tumor cells located in the sinusoids and 7.1 +/- 0.9% of tumor cells in the TPVs. The NO synthase inhibitor N(G)-nitro-L-arginine methyl ester completely blocked the NO burst (P < 0.001) and inhibited the apoptosis of B16F1 cells in the sinusoids by 77%. However, the rate of tumor cell apoptosis in the TPVs was not changed. There were 5-fold more metastatic nodules in the livers of N(G)-nitro-L-arginine methyl ester-treated mice (P < 0.05). The inactive enantiomer N(G)-nitro-D-arginine methyl ester had no effect on the initial NO burst or on apoptosis of tumor cells in vivo. Both annexin V phosphatidylserine plasma membrane labeling and DNA end labeling of apoptotic cells were demonstrated after a 5-min exposure (a time equivalent to the initial transient NO induction in vivo) of B16F1 cells to a NO donor in vitro. These results identify the existence of a natural defense mechanism against cancer metastasis whereby the arrest of tumor cells in the liver induces endogenous NO release, leading to sinusoidal tumor cell killing and reduced hepatic metastasis formation. PMID- 11059785 TI - Caveolin-1 levels are down-regulated in human colon tumors, and ectopic expression of caveolin-1 in colon carcinoma cell lines reduces cell tumorigenicity. AB - Caveolin-1 expression and function were investigated in human colon cancer. Low levels of caveolin-1 mRNA and protein were detected in several colon carcinoma cell lines. Moreover, caveolin-1 protein levels were significantly reduced in human tumor epithelial mucosa (3.6 +/- 1.4-fold) when compared with normal colon mucosa for a majority (10 of 15) of the patients characterized. To directly assess the role of caveolin-1 in tumor development, caveolin-1 was reexpressed in the HT29 and DLD1 colon carcinoma cells, and the resulting HT29-cav-1 or DLD1-cav 1 cells were tested for tumorigenicity in nude mice. In most experiments, tumor formation was either blocked or retarded for HT29-cav-1 cells (10 of 13 mice) and DLD1-cav-1 cells (5 of 7 mice), as compared with both mock-transfected and parental HT29 or DLD1 cells. Interestingly, basal caveolin-1 levels were significantly reduced in HT29-cav-1 and DLD1-cav-1 cells isolated from tumors. Likewise, endogenous caveolin-1 mRNA and protein levels were found to be reduced in NIH-3T3 cells recovered from tumors after injection into nude mice. Thus, reexpression of caveolin-1 in colon carcinoma lines reduced the probability of tumor formation in vivo, and when tumors did develop from either HT29-cav-1, DLD1 cav-1, or NIH-3T3 cells, lower basal levels of caveolin-1 were detected. Finally, evidence was obtained indicating that initial caveolin-1 down-regulation in colon cancer cells need not be an entirely irreversible process because cell survival on selection for either drug resistance or increased metastatic potential correlated with increased caveolin-1 expression levels. PMID- 11059786 TI - Epidermal growth factor receptor transcriptionally up-regulates vascular endothelial growth factor expression in human glioblastoma cells via a pathway involving phosphatidylinositol 3'-kinase and distinct from that induced by hypoxia. AB - Glioblastomas are highly vascular malignant brain tumors that often overexpress vascular endothelial growth factor (VEGF). They also frequently overexpress epidermal growth factor receptor (EGFR) and contain regions of hypoxia, both conditions that can induce VEGF. We examined VEGF regulation in U87 MG human glioblastoma cells and in U87/T691 cells, a clonal derivative that contains a truncated erbB2/Neu receptor that interferes with EGFR signaling through the formation of nonfunctional heterodimeric receptor complexes. U87/T691 cells contained approximately one-half as much VEGF mRNA as did U87 MG cells under normoxic conditions (21% oxygen). Pharmacological inhibition of EGFR, Ras, or PI(3) kinase, but not MAP kinase, led to a significant decrease in VEGF mRNA levels in U87 MG cells. VEGF promoter activity in transient transfections was decreased by either pharmacological or genetic inhibition of EGFR, Ras, or phosphatidylinositol 3'-kinase [PI(3) kinase]. However, inhibition of PI(3) kinase or EGFR did not completely abolish induction of VEGF mRNA by hypoxia (0.2% oxygen). Likewise, VEGF mRNA expression was induced 3-fold by hypoxia in EGFR inhibited U87/T691 cells, comparable with the fold induction seen in parental U87 MG cells, although the absolute level of message under hypoxia was higher in U87 MG cells. In transient transfections, a luciferase reporter construct containing a 1.2-kb fragment of the VEGF promoter, lacking the known hypoxic-responsive element (HRE), showed up-regulation after EGF stimulation to the same degree as the full-length, 1.5-kb VEGF promoter construct retaining the HRE. Furthermore, activity of the HRE-deleted, 1.2-kb promoter luciferase reporter was down regulated by PI(3) kinase inhibition. Therefore, in glioblastoma cells, transcriptional regulation of the VEGF promoter by EGFR appears to involve Ras/PI(3) kinase and to be distinct from signals induced by hypoxia. PMID- 11059787 TI - Inhibition of HER2/neu (erbB-2) and mitogen-activated protein kinases enhances tamoxifen action against HER2-overexpressing, tamoxifen-resistant breast cancer cells. AB - HER2/neu (erbB-2) overexpression has been causally associated with tamoxifen resistance in human breast cancer cells. Forced expression of HER2 in MCF-7 breast cancer cells resulted in mitogen-activated protein kinase (MAPK) hyperactivity and tamoxifen resistance. Inhibition of HER2 and MAPKs with AG1478 and U0126, respectively, as well as dominant-negative MEK-1/2 constructs restored the inhibitory effect of tamoxifen on estrogen receptor (ER)-mediated transcription and cell proliferation. Both AG1478 and U0126 also restored the tamoxifen-mediated association of ER with nuclear receptor corepressor (N-CoR) in the antiestrogen-resistant MCF-7 cells. Treatment with a combination of tamoxifen and a HER2 kinase inhibitor reduced tumor MAPK activity and markedly prevented growth of HER2-overexpressing MCF-7 xenografts in athymic mice. Thus, blockade of HER2 and MAPK signaling may enhance tamoxifen action and abrogate antiestrogen resistance in human breast cancer. PMID- 11059788 TI - A modified p53 overcomes mdm2-mediated oncogenic transformation: a potential cancer therapeutic agent. AB - The antiproliferative activities of wild-type (wt) p53 are inhibited by mdm2 (murine double minute2) oncogene product. We tested growth suppression activity of p53 14/19, an engineered p53 variant, which does not bind mdm2 and is completely resistant to the inhibition by mdm2. p53 14/19, unlike wt p53, suppressed the growth of cancer cells that contain amplified mdm2 oncogene efficiently by direct DNA transfection or adenovirus-mediated gene transfer. In addition, p53 14/19 also inhibited the growth of several different cancer cell lines expressing low levels of mdm2 oncogene product as efficiently as wt p53. We further examined the antioncogenic potencies of p53 14/19 in the rat embryo fibroblast cotransformation assay. Addition of wt p53 failed to cause any significant decrease in ras plus mdm2 foci counts. In contrast, cotransfection of p53 14/19 with ras and mdm2 significantly reduced foci number. In similar experiments, cotransfection of wt p53 or 14/19 p53 resulted in significant inhibition of oncogenic transformation in rat embryo fibroblast mediated by an activated ras plus c-myc, adenovirus E1A, or human papillomavirus E7 oncogenes. Therefore, these results suggest that p53 14/19 modified tumor suppressor gene may be a promising therapeutic agent for human cancers that express abnormally high levels of mdm2 oncogene product. PMID- 11059789 TI - Antiserum raised against an epitope of the cholecystokinin B/gastrin receptor inhibits hepatic invasion of a human colon tumor. AB - Serum gastrin is known to be elevated in patients with liver-metastasizing colon cancer; thus, cholecystokinin (CCK) B/gastrin receptors may also be up-regulated. A liver-invasive model of colon cancer was established with the human colonic cell line C170HM2, which expresses the CCKB/gastrin receptor at both the gene and protein level. An antiserum has been derived that is directed against the NH2 terminal 17 amino acids of the human CCKB/gastrin receptor coupled to diphtheria toxoid. The peptide was denoted gastrin receptor protein (GRP) 1. The therapeutic effect of GRP1 antiserum was evaluated on the liver invasion of C170HM2 tumors. Biodistribution studies revealed that GRP1 antiserum localized preferentially within the liver tumors when compared with normal liver tissue (1.5-fold increase after 24 h; P < 0.05). Antiserum against GRP1 inhibited both tumor take rate and final liver tumor weight when compared with treatment with control serum in mice with an increasing tumor burden. Liver tumor weights were reduced from 0.37 to 0.10 gram (P = 0.0155), 1.25 grams to 0.76 gram (P = 0.003) and 1.89 grams to 0.76 gram (P = 0.0068, all Mann-Whitney nonparametric U test). Necrosis and apoptosis were increased in the GRP1 antiserum-treated tumors when compared with control serum-treated tumors. As shown by Western blotting, CCKB/gastrin receptor expression of C170HM2 xenografts after treatment with GRP1 antiserum shifted to a predominantly lower molecular weight form (Mr 45,000) that is known to be an internalized form of the receptor. In conclusion, targeting of the CCKB/gastrin receptor may yield a valuable therapeutic modality for the treatment of advanced colon cancer. PMID- 11059790 TI - The endosomal-lysosomal system of neurons in Alzheimer's disease pathogenesis: a review. AB - A prominent feature of brain pathology in Alzheimer's disease is a robust activation of the neuronal lysosomal system and major cellular pathways converging on the lysosome, namely, endocytosis and autophagy. Recent studies that identify a disturbance of the endocytic pathway as one of the earliest known manifestation of Alzheimer's disease provide insight into how beta amyloidogenesis might be promoted in sporadic Alzheimer's disease, the most prevalent and least well understood form of the disease. Primary lysosomal dysfunction has historically been linked to neurodegeneration. New data now directly implicate cathepsins as proteases capable of initiating, as well as executing, cell death programs in certain pathologic states. These and other studies support the view that the progressive alterations of lysosomal function observed during aging and Alzheimer's disease contribute importantly to the neurodegenerative process in Alzheimer's disease. PMID- 11059792 TI - Cholinesterase inhibitors stabilize Alzheimer disease. AB - During the last decade, a systematic effort to develop a pharmacological treatment for Alzheimer disease (AD) has resulted into three drugs being registered for the first time in USA and Europe for this specific indication. All three are cholinesterase inhibitors (ChEI). The major therapeutic effect of ChEI on AD patients is to maintain cognitive function at a constant level during a 6 months to one year period of treatment as compared to placebo. Additional drug effects might be slowing cognitive deterioration and improving behavioral and daily living conditions. Comparison of clinical effects of 6 ChEI demonstrates a rather similar magnitude of improvement in cognitive measures. For some drugs. this may represent an upper limit while for other it may still be possible to increase further the benefit. In order to maximize and prolong positive drug effects it is important to start early and adjust dosage during the treatment. Recent studies show that in many patients the stabilization effect produced by ChEI can be prolonged for as long as a 24 month period. In order to explain the stabilizing effect of ChEI, a mechanism other than AChE inhibition, based on beta amyloid metabolism, is postulated. PMID- 11059791 TI - Neuroinflammatory signaling upregulation in Alzheimer's disease. AB - Alzheimer's disease (AD) is a progressive, neurodestructive process of the human neocortex, characterized by the deterioration of memory and higher cognitive function. A progressive and irreversible brain disorder, AD is characterized by three major pathogenic episodes involving (a) an aberrant processing and deposition of beta-amyloid precursor protein (betaAPP) to form neurotoxic beta amyloid (betaA) peptides and an aggregated insoluble polymer of betaA that forms the senile plaque, (b) the establishment of intraneuronal neuritic tau pathology yielding widespread deposits of agyrophilic neurofibrillary tangles (NFT) and (c) the initiation and proliferation of a brain-specific inflammatory response. These three seemingly disperse attributes of AD etiopathogenesis are linked by the fact that proinflammatory microglia, reactive astrocytes and their associated cytokines and chemokines are associated with the biology of the microtubule associated protein tau, betaA speciation and aggregation. Missense mutations in the presenilin genes PS1 and PS2, implicated in early onset familial AD, cause abnormal betaAPP processing with resultant overproduction of betaA42 and related neurotoxic peptides. Specific betaA fragments such as betaA42 can further potentiate proinflammatory mechanisms. Expression of the inducible oxidoreductase cyclooxygenase-2 and cytosolic phospholipase A2 (cPLA2) are strongly activated during cerebral ischemia and trauma, epilepsy and AD, indicating the induction of proinflammatory gene pathways as a response to brain injury. Neurotoxic metals such as aluminum and zinc, both implicated in AD etiopathogenesis, and arachidonic acid, a major metabolite of brain cPLA2 activity, each polymerize hyperphosphorylated tau to form NFT-like bundles. Further, epidemiological and longitudinal studies have identified a reduced risk for AD in patients (<70 yrs) previously treated with non-steroidal anti-inflammatory drugs for non-CNS afflictions that include arthritis. This review will focus on the interrelationships between the mechanisms of PS1, PS2 and betaAPP gene expression, tau and betaA deposition and the induction, regulation and proliferation in AD of the neuroinflammatory response. Novel therapeutic interventions in AD are discussed. PMID- 11059793 TI - Increased calpain expression is associated with apoptosis in rat spinal cord injury: calpain inhibitor provides neuroprotection. AB - Calpain content was investigated in the lesion of rat spinal cord at 1, 4, 24, and 72 h following injury induced by the weight-drop (40 g-cm force) technique. Calpain content was increased in the lesion, and was highest at 24 h following injury. microCalpain mRNA level in the lesion was increased by 58.4% (p = 0.0135) at 24 h following trauma, compared to sham. Alterations in mRNA expression in the lesion increased bax/bcl-2 ratio by 20.8% (p = 0.0395) at this time point, indicating a commitment to apoptosis. Therapeutic effect of the calpain inhibitor E-64-d (1 mg/kg) was studied in SCI rats following administration for 24 h. Internucleosomal DNA fragmentation (apoptosis) was observed in SCI rats, but not in sham or E-64-d treated rats. These results indicate a new information that E 64-d has the therapeutic potential for inhibiting apoptosis in SCI. PMID- 11059794 TI - 5-HT-HPA interactions in two models of transgenic mice relevant to major depression. AB - Reciprocal interactions between central 5-HT system and hypothalamo-pituitary adrenal (HPA) axis are of particular relevance with regard to depression, in which alterations of both systems have been evidenced. In order to further explore these interactions, two models of mutant mice have been used. They consisted of knock-out mice lacking the 5-HT transporter (5-HTT-/-) and of transgenic mice with impaired glucocorticoid receptor (GR-i) expression. Under control conditions. the functional properties of 5-HT(1A) autoreceptors in GR-i mice were as in their paired wild-type. However, both chronic stress and long term treatment with fluoxetine induced abnormal adaptive changes in 5-HT(1A) autoreceptor functioning in GR-i mice. On the other hand, a marked desensitization of 5-HT(1A) autoreceptors was found in 5-HTT-/- mice as compared with paired wild-type animals, and this phenomenon was further enhanced by exposure to stressful conditions. These data show that alterations of HPA axis at the gene level has consequences on 5-HT neurotransmission, and reciprocally, that 5-HTT knock-out affects HPA-dependent responses to stress. PMID- 11059795 TI - New neurochemical markers for psychosis: a working hypothesis of their operation. AB - Reelin (Reln) is expressed in specific GABAergic neurons in layer I and II of neocortex, and is secreted into the extracellular matrix where it surrounds dendrites, spines and neurite arborizations, and binds to integrin receptors located on post-synaptic densities of apical dendritic spines. Experiments in rodents (including wild type or reeler heterozygous mice) and non-human primates suggest the Reln secreted in the extracellular matrix of neocortex, via integrin receptors, modulates the function of the adaptor protein DAB1(drosophila disable gene) homologous product) thereby participating in dynamic processes associated with plasticity changes in dendrites, dendritic spines and their synapses. A local protein synthesis at dendritic spines (ie the activity regulated cytoskeleton associated protein, Arc) probably acts as a signal for plastic modulatory activities in synapses operative in neural group interactions. A research strategy directed toward identifying specific neurochemical markers operative in the etiopathology of psychotic disorders lead to the identification of a downregulation (30-50%) of Reln and glutamic acid decarboxylase 67(GAD67) expression in prefrontal cortex and other brain areas of schizoprenia and bipolar disorder patients with psychosis. These downregulations were not due to neuronal damage, postmortem interval, or antipsychotic medication. The dysfunction of GABAergic interneurons observed in psychotic brains in combination with reduced Reln expression and downregulation of Reln-integrin receptor interaction, may provide an explanation for the reported decrease in neuropile expression including dendritic spine density reduction, in neocortex of schizophrenia patients. This downregulation of neuropile plasticity may be a factor to be considered in the etiology of the disintegration of consciousness, which is one of the primary signs of psychosis. PMID- 11059796 TI - Allostasis, allostatic load, and the aging nervous system: role of excitatory amino acids and excitotoxicity. AB - The adaptive responses of the body to challenges, often known as "stressors", consists of active responses that maintain homeostasis. This process of adaptation is known as "allostasis", meaning "achieving stability through change". Many systems of the body show allostasis, including the autonomic nervous system and hypothalamo-pituitary-adrenal (HPA) axis and they help to re establish or maintain homeostasis through adaptation. The brain also shows allostasis, involving the activation of nerve cell activity and the release of neurotransmitters. When the individual is challenged repeatedly or when the allostatic systems remain turned on when no longer needed, the mediators of allostasis can produce a wear and tear on the body that has been termed "allostatic load". Examples of allostatic load include the accumulation of abdominal fat, the loss of bone minerals and the atrophy of nerve cells in the hippocampus. Circulating stress hormones play a key role, and, in the hippocampus, excitatory amino acids and NMDA receptors are important mediators of neuronal atrophy. The aging brain seems to be more vulnerable to such effects, although there are considerable individual differences in vulnerability that can be developmentally determined. Yet, at the same time, excitatory amino acids and NMDA receptors mediate important types of plasticity in the hippocampus. Moreover, the brain retains considerable resilience in the face of stress, and estrogens appear to play a role in this resilience. This review discusses the current status of work on underlying mechanisms for these effects. PMID- 11059797 TI - Neurochemical and molecular pharmacological aspects of the GABA(B) receptor. AB - Metabotropic gamma-aminobutyric acid (GABA)B receptors are known to modulate the synaptic release of various neurotransmitters in the nervous system. Activation of GABA(B) receptor induces the inhibition of adenylyl cyclase activity, while it does not stimulate the formation of inositol phosphates. Activation of a potassium conductance and suppression of a calcium conductance are also recognized, similarly to some of G protein-coupled receptors. Recent molecular cloning has revealed that GABA(B) receptor possesses a large extracellular domain including the binding site for GABA and seven transmembrane domains. Their molecular structures in the brain are unique and interesting because of heterodimerization consisting of two distinct genes: GABABR1 and GABABR2. Such assembled receptors can be classified as a novel type of the metabotropic receptor superfamily. PMID- 11059798 TI - Pharmacological and functional characterization of astrocytic GABA transport: a short review. AB - GABA neurotransmission is terminated by high affinity transport mediated by a number of carriers on neurons and astrocytes. So far four different carriers have been cloned and their cellular distribution has been partly worked out. It is generally believed that GAT-1 (mouse homologue GAT1) is the quantitatively most important of the transporters and it is primarily present on GABAergic neurons but also to some extent on astrocytes. The pharmacological properties of neuronal and astrocytic GABA uptake have been studied extensively and recently the GABA analogue N-methyl-Exo-THPO has been reported to act as a selective and potent (IC50 28 microM) astroglial GABA transport inhibitor with a 15-fold selectivity. It has moreover been reported to act as an anticonvulsant in animal models of epilepsy. This may underline the functional importance of astrocytic GABA uptake in relation to seizure activity. PMID- 11059799 TI - Regulation of AMPA receptors by phosphorylation. AB - The AMPA receptors for glutamate are oligomeric structures that mediate fast excitatory responses in the central nervous system. Phosphorylation of AMPA receptors is an important mechanism for short-term modulation of their function, and is thought to play an important role in synaptic plasticity in different brain regions. Recent studies have shown that phosphorylation of AMPA receptors by cAMP-dependent protein kinase (PKA) and Ca2+- and calmodulin-dependent protein kinase II (CaMKII) potentiates their activity, but phosphorylation of the receptor subunits may also affect their interaction with intracellular proteins, and their expression at the plasma membrane. Phosphorylation of AMPA receptor subunits has also been investigated in relation to processes of synaptic plasticity. This review focuses on recent advances in understanding the molecular mechanisms of regulation of AMPA receptors, and their implications in synaptic plasticity. PMID- 11059800 TI - Molecular signals for development of neuronal circuitry in the retina. AB - In this review article, we summarize recently accumulated knowledge regarding the molecular mechanisms, which control retinal development. Retinal neurons are born in two waves of cytogenesis. In the first wave, neurons of cone circuitry are generated, whereas in the second wave, rod circuitry is added. Neurons generated in these two waves of cytogenesis differ in many respects, including the molecular cues used for migrational guidance. The neurons generated in the second phase of proliferation are arranged in radial columns associated with Muller cells, whereas those of the first phase are often found outside the radial columns. Certain early born cone photoreceptors may form templates for the arrangement of additional mosaics of other cell types. These mosaic arrangements of cell bodies are subsequently refined by lateral displacement of cells and apoptosis. Muller cells may play an important role in directing migration of second phase neurons within groups of radial columns and also in guiding the projections of these neurons so that specific connections are formed. The Muller cell's ability to exert these influences perhaps resides in a variety of cell adhesion molecules such as L1/NgCAM, F11, and 5A11, which are expressed on the surface of Muller cells and retinal neurons. CAMs also promote neurite outgrowth through second messenger pathways. PMID- 11059801 TI - Synaptosomes still viable after 25 years of superfusion. AB - Superfused synaptosomes have been utilized in studies of neurotransmitter release during 25 years. This review summarizes the aspects of neurotransmission that have been and could be successfully investigated with this technique. The major aim of the article is to draw attention on the versatility of superfused synaptosomes and to suggest how the system could be exploited in clarifying several aspects of synaptic neurochemistry including neurotransmitter transport, receptor localization, receptor-receptor interactions, functional aspects of multi-sited receptor complexes, receptor heterogeneity and mechanisms of neurotransmitter exocytosis-endocytosis. PMID- 11059802 TI - Synaptic vesicle proteins and neuronal plasticity in adrenergic neurons. AB - The neurons in the superior cervical ganglion are active in plasticity and re modelling in order to adapt to requirements. However, so far, only a few studies dealing with synaptic vesicle related proteins during adaptive processes have been published. In the present paper, changes in content and expression of the synaptic vesicle related proteins in the neurons after decentralization (cutting the cervical sympathetic trunk) or axotomy (cutting the internal and external carotid nerves) were studied. Immunofluorescence studies were carried out using antibodies and antisera against integral membrane proteins, vesicle associated proteins, NPY, and the enzymes TH and PNMT. For colocalization studies, the sections were simultaneously double labelled. Confocal laser scanning microscopy was used for colocalization studies as well as for semi-quantification analysis, using the computer software. Westen blot analysis, in situ 3'-end DNA labelling, and in situ hybridization were also employed. After decentralization of the ganglia several of the synaptic vesicle proteins (synaptotagmin I, synaptophysin, SNAP-25, CLC and GAP-43) were increased in the iris nerve terminal network, but with different time patterns, while TH-immunoreactivity had clearly decreased. In the ganglia, these proteins had decreased at 1 day after decentralization, probably due to degeneration of the pre-ganglionic nerve fibres and terminals. At later intervals, these proteins, except SNAP-25, had increased in the nerve fibre bundles and re-appeared in nerve fibres outlining the principal neurons. PMID- 11059803 TI - Signaling events during swelling and regulatory volume decrease. AB - Brain cell swelling compromises neuronal function and survival by the risk of generation of ischemia episodes as compression of small vessels occurs due to the limits to expansion imposed by the rigid skull. External osmolarity reductions or intracellular accumulation of osmotically active solutes result in cell swelling which can be counteracted by extrusion of osmolytes through specific efflux pathways. Characterization of these pathways has received considerable attention, and there is now interest in the understanding of the intracellular signaling events involved in their activation and regulation. Calcium and calmodulin, phosphoinositides and cAMP may act as second messengers, carrying the information about a cell volume change into signaling enzymes. Small GTPases, protein tyrosine kinases and phospholipases, also appear to be part of the signaling cascades ultimately modulating the osmolyte efflux pathways. This review focus on i) the influence of hyposmotic and isosmotic swelling on these signaling events and molecules and ii) the effects of manipulating their function on the osmolyte fluxes, particularly K+, CI- and amino acids, and on the consequent efficiency of cell volume adjustment. PMID- 11059805 TI - Neurochemistry of brain neuroendocrine immune system: signal molecules. AB - The aim of this review is not so much to show the problem of neuroendocrine, neurophysiologic, and neurochemical mechanisms of the immune system regulation of the organism by brain (there is a great deal of literature about it), as to solve the problem of whether the brain itself is an immune organ, and also to define cellular, neurochemical, and immunological properties of the brain for its immune defense when the blood-brain barrier is not damaged in spite of the penetration of the infection to brain. The accumulated literary data on CNS interaction with the immune system, expression of several cytokines and their receptors in the neurons of human brain culture, in astrocytes and microglia, all testify to the existence of a brain immune system. Recently studies appeared on the expression of major histocompatibility complex in brain neurons. It does not exclude the possibility of expression of immunoglobulins (or immunoglobulin-like proteins) in brain cells. Data obtained by us on the biosynthesis of a number of known interleukins and new cytokines in neurosecretory neurons of hypothalamus (N. Paraventricularis and N. Supraopticus) demonstrate that neuroendocrine nuclei of the hypothalamus are the center for neuroendocrine and immune systems of brain. PMID- 11059806 TI - Isoprostanes, novel markers of oxidative injury, help understanding the pathogenesis of neurodegenerative diseases. AB - Isoprostanes are prostaglandin-like compounds which are formed by free radical catalysed peroxidation of arachidonic acid esterified in membrane phospholipids. They are emerging as a new class of sensitive, specific and reliable markers of in vivo lipid peroxidation and oxidative damage. Since their initial description of in 1990, the rapid development of analytical methods for isoprostane measurement has allowed to overcome some of the pitfalls of the previous and most widely used methods of assessing free radical injury. Here, we summarise the current knowledge on these novel class lipid peroxidation products and the advantages of monitoring their formation to better define the involvement of oxidative stress in neurological diseases. Although the literature data are still not abundant, they indicate that in vivo or post mortem cerebrospinal fluid and brain tissue levels of isoprostane are increased in some diseases such as multiple sclerosis, Alzheimer's disease, Huntington's disease, and Creutzfeldt Jakob disease. PMID- 11059804 TI - NO synthase and NO-dependent signal pathways in brain aging and neurodegenerative disorders: the role of oxidant/antioxidant balance. AB - Nitric oxide and other reactive nitrogen species appear to play several crucial roles in the brain. These include physiological processes such as neuromodulation, neurotransmission and synaptic plasticity, and pathological processes such as neurodegeneration and neuroinflammation. There is increasing evidence that glial cells in the central nervous system can produce nitric oxide in vivo in response to stimulation by cytokines and that this production is mediated by the inducible isoform of nitric oxide synthase. Although the etiology and pathogenesis of the major neurodegenerative and neuroinflammatory disorders (Alzheimer's disease, amyothrophic lateral sclerosis, Parkinson's disease, Huntington's disease and multiple sclerosis) are unknown, numerous recent studies strongly suggest that reactive nitrogen species play an important role. Furthermore, these species are probably involved in brain damage following ischemia and reperfusion, Down's syndrome and mitochondrial encephalopathies. Recent evidence also indicates the importance of cytoprotective proteins such as heat shock proteins (HSPs) which appear to be critically involved in protection from nitrosative and oxidative stress. In this review, evidence for the involvement of nitrosative stress in the pathogenesis of the major neurodegenerative/ neuroinflammatory diseases and the mechanisms operating in brain as a response to imbalance in the oxidant/antioxidant status are discussed. PMID- 11059807 TI - MR spectroscopy: a powerful tool for investigating brain function and neurological diseases. AB - Magnetic resonance spectroscopy (MRS) has attracted much attention in recent years and has become an important tool to study in vivo particular biochemical aspects of brain disorders. Since the proton is the most sensitive stable nucleus for MRS, and since almost all metabolites contain hydrogen atoms, investigation by in vivo 1H MRS provides chemical information on tissue metabolites, thus enabling a non-invasive assessment of changes in brain metabolism underlying several brain diseases. In this review a brief description of the basic principles of MRS is given. Moreover, we provide some explanations on the techniques and technical problems related to the use of 1H MRS in vivo including water suppression, localization, editing, quantitation and interpretation of 1H spectra. Finally, we discuss the more recent advancement in three major areas of neurological diseases: brain tumors, multiple sclerosis, and inborn errors of metabolism. PMID- 11059808 TI - One path to cell death in the nervous system. AB - Both acute and chronic insults to the nervous system can result in changes in homeostasis that result in cell death or recovery processes that alter function. The signaling mechanisms for this broad spectrum of events that impair neurological function span the gamut from abrupt injury to the slow onset of neurodegenerative diseases in extreme aging. A common element in all of these events is the triggering of signal cascades that determine cellular commitment to apoptosis as a ameliorative alternative to inflammatory necrosis. Key in these cascades is the activation of the caspase and Bcl-family of proteins by the NF kappaB transcription factor. Here we consider aspects of specificity of activation as a result of the differential expression of NF-kappaB proteins and their regulation of selective genes as a result of binding to select DNA consensus sequences out of the 64 different combinations that constitute the NF kappaB DNA binding consensus sequence. PMID- 11059809 TI - 31P-MRS-based determination of brain intracellular and interstitial pH: its application to in vivo H+ compartmentation and cellular regulation during hypoxic/ischemic conditions. AB - In the last decade, significant progress has been made in the characterization of pH regulation in nervous tissue in vitro. However, little work has been directed at understanding how pH regulatory mechanisms function in vivo. We are interested in how ischemic acidosis can effect pH regulation and modulate the extent of post ischemic brain damage. We used 31P-MRS to determine normal in vivo pH(i) and pH(e) simultaneously in both the isolated canine brain and the intact rat brain. We observed that the 31P(i) peak in the 31P-MRS spectrum is heterogeneous and can be deconvoluted into a number of discrete constituent peaks. In a series of experiments, we identified these peaks as arising from either extracellular or intracellular sources. In particular, we identified the peak representing the neurons and astrocytes and showed that they maintain different basal pH (6.95 and 7.05, respectively) and behave differently during hypoxic/ischemic episodes. PMID- 11059810 TI - Mechanisms of L-cysteine neurotoxicity. AB - We review here the possible mechanisms of neuronal degeneration caused by L cysteine, an odd excitotoxin. L-Cysteine lacks the omega carboxyl group required for excitotoxic actions via excitatory amino acid receptors, yet it evokes N methyl-D-aspartate (NMDA) -like excitotoxic neuronal death and potentiates the Ca2+ influx evoked by NMDA. Both actions are prevented by NMDA antagonists. One target for cysteine effects is thus the NMDA receptor. The following mechanisms are discussed now: (1) possible increase in extracellular glutamate via release or inhibition of uptake/degradation, (2) generation of cysteine alpha-carbamate, a toxic analog of NMDA, (3) generation of toxic oxidized cysteine derivatives, (4) chelation of Zn2+ which blocks the NMDA receptor-ionophore, (5) direct interaction with the NMDA receptor redox site(s), (6) generation of free radicals, and (7) formation of S-nitrosocysteine. In addition to these, we describe another new alternative for cytotoxicity: (8) generation of the neurotoxic catecholamine derivative, 5-S-cysteinyl-3,4-dihydroxyphenylacetate (cysdopac). PMID- 11059811 TI - Phosphate-activated glutaminase and mitochondrial glutamine transport in the brain. AB - A review of the properties of purified and tissue bound phosphate activated glutaminase (PAG) in brain and kidney (pig and rat) is presented, based on kinetic, electron microscopic and immunocytochemical studies. PAG is a mitochondrial enzyme and two pools can be separated, a soluble and membrane associated one. Intact mitochondria appear to express PAG accessible only to the outer phase of the inner mitochondrial membrane. This PAG has properties similar to that of the membrane fraction and polymeric form of purified enzyme. PAG in the soluble fraction has properties similar to that of the monomeric form of purified enzyme and is assumed to be dormant due to the high matrix concentration of the inhibitor glutamate. A hypothetical model for the localization of PAG in the mitochondria is presented. The activity of PAG in vivo is assumed to be regulated by cytosolic glutamate and other compounds, that affect the activation by phosphate. Glutamine is transported into brain and kidney mitochondria by a protein catalyzed energy requiring process, which may be mediated by more than one protein. There is no correlation between glutamine hydrolysis and transport. PMID- 11059813 TI - William Hogarth, unwitting neurochemist? AB - William Hogarth's famous etching Gin Lane is often used to illustrate the debilitating results of alcohol addiction. Less well known is the companion etching Beer Street in which death, murder and squalor are replaced by health, orderliness and joy. Some 250 years later, the rise of science, and specifically of neurochemical research, has defined how the malnutrition, including avitaminosis, resulting from addiction to distilled spirits (rather than more judicious use of less potent alcoholic beverages) disturbs brain metabolism and function. These two etchings, which have survived for their historical and artistic value, continue to have sociological and clinical relevance. PMID- 11059812 TI - How many endobains are there? AB - Oxidative metabolism is very active in brain, where large amounts of chemical energy as ATP molecules are consumed, mostly required to maintain cellular Na+/K+ gradients through the participation of the sodium pump (Na+,K+-ATPase), whose activity is selectively and potently inhibited by the alkaloid ouabain. Na+/K+ gradients are involved in nerve impulse propagation, in neurotransmitter release and cation homeostasis in the nervous system. Likewise, enzyme activity modulation is crucial for maintaining normal blood pressure and cardiovascular contractility as well as renal sodium excretion. The present article reviews the progress in disclosing putative ouabain-like substances, examines their denomination according to different research teams, tissue or biological fluid sources, extraction and purification, assays, biological properties and chemical and biophysical features. When data is available, comparison with ouabain itself is mentioned. Likewise, their potential action in normal physiology as well as in experimental and human pathology is summarized. PMID- 11059814 TI - A journey in the neurosciences from 1950 to 2000. PMID- 11059815 TI - Glial fibrillary acidic protein: GFAP-thirty-one years (1969-2000). AB - It is now well established that the glial fibrillary acidic protein (GFAP) is the principal 8-9 nm intermediate filament in mature astrocytes of the central nervous system (CNS). Over a decade ago, the value of GFAP as a prototype antigen in nervous tissue identification and as a standard marker for fundamental and applied research at an interdisciplinary level was recognized (Raine, 135). As a member of the cytoskeletal protein family, GFAP is thought to be important in modulating astrocyte motility and shape by providing structural stability to astrocytic processes. In the CNS of higher vertebrates, following injury, either as a result of trauma, disease, genetic disorders, or chemical insult, astrocytes become reactive and respond in a typical manner, termed astrogliosis. Astrogliosis is characterized by rapid synthesis of GFAP and is demonstrated by increase in protein content or by immunostaining with GFAP antibody. In addition to the major application of GFAP antisera for routine use in astrocyte identification in the CNS, the molecular cloning of the mouse gene in 1985 has opened a new and rich realm for GFAP studies. These include antisense, null mice, and numerous promoter studies. Studies showing that mice lacking GFAP are hypersensitive to cervical spinal cord injury caused by sudden acceleration of the head have provided more direct evidence for a structural role of GFAP. While the structural function of GFAP has become more acceptable, the use of GFAP antibodies and promoters continue to be valuable in studying CNS injury, disease, and development. PMID- 11059816 TI - Was the ASN a myelin society? American Society for Neurochemistry. PMID- 11059817 TI - Antiretroviral therapy 2000. AB - As we enter the new millennium, there have been dramatic improvements in the care of patients with HIV infection. These have prolonged life and decreased morbidity and mortality. There are fourteen currently available antiretrovirals approved in the United States for the treatment of this infection. The medications, including their pharmacokinetic properties, side effects, and dosing are reviewed. In addition, the current approach to the use of these medicines is discussed. PMID- 11059818 TI - Total synthesis of a demethoxy-egonol from Styrax obassia. AB - The total synthesis of a demethoxy-egonol isolated from Styrax obassia, 5-(3 hydroxypropyl)-2-(3',4'-methylenedioxyphenyl)benzofuran (9), is described. The key steps involve the construction of a 2-arylbenzofuran skeleton 7 from methyl 3 (4-hydroxyphenyl)propionate with 2-chloro-2-methylthio-(3',4' methylenedioxy)acetophenone (6) in the presence of ZnCl2 and successive desulfurization of the resulting product 7. PMID- 11059819 TI - Determination of terbutaline enantiomers in human plasma by coupled achiral chiral high performance liquid chromatography. AB - Achiral-chiral column switching HPLC assay was developed to allow the separation and quantification of the enantiomers of terbutaline in human plasma by means of fluorescence detection. Plasma samples were prepared by solid-phase extraction with sep-pak silica, followed by HPLC assay. The enantiomers of terbutaline and the internal standard were separated from the biological matrix on a silica column, and the two enantiomers were resolved and quantified on a Sumichiral OA 4900 column. The two columns were connected by a switching valve equipped with silica trap column. The trap column was used to concentrate the terbutaline in the eluent from the achiral column before back flushing onto the chiral phase. For each enantiomers, the assay was linear between 2.5-125 ng/ml (r=0.9999) and detection limit was 1.0 ng/ml. PMID- 11059820 TI - Synthesis of heterocyclic quinones containing bridgehead nitrogen atom from 2 aminonaphtho[2,3-d]thiazole-4,9-dione. AB - Imidazonaphthothiazole derivatives 3-6 were prepared by treatment of 2 aminonaphtho[2,3-d]-thiazole-4,9-dione(1) with phenacyl bromide, chloroacetic acid, diethyl oxalate and 2,3-dichloroquinoxaline respectively. The reaction of 1 with ethyl acrylate, ethyl acetoacetate and diethyl malonate gave the corresponding naphthothiazolopyrimidine derivatives 8-11. PMID- 11059821 TI - Synthesis and in vitro cytotoxicity of 2-alkylaminosubstituted quinoline derivatives. AB - Eight 2-alkylaminosubstituted 5,8-dimethoxy-4-methylquinolines and nine 2 alkylaminosubstituted or 2,6-disubstituted 4-methylquinoline-5,8-diones were synthesized and evaluated in vitro cytotoxicity against four human cancer cell lines (HOP62, SK-OV-3, HCT15 and SF295). PMID- 11059822 TI - Antioxidative phenolic compounds from the roots of Rhodiola sachalinensis A. Bor. AB - The acetone extract of the roots of Rhodiola sachalinensis has furnished six phenolic compounds which exhibited significant scavenging effects against DPPH free radical. The structures of these compounds were identified and determined as gallic acid (1), (-)-epigallocatechin 3-O-gallate (2), kaempferol (3), kaempferol 7-O-alpha-L-rhamnopyranoside (4), herbacetin 7-O-alpha-L-rhamnopyranoside, (5) and rhodiolinin (6) by physico-chemical and spectral evidences. PMID- 11059823 TI - Spirobenzylisoquinoline alkaloids from Corydalis ochotensis. AB - Separation of the alkaloids from the aerial parts of Corydalis ochotensis afforded a new spirobenzylisoquinoline alkaloid, 8-O-acetylcorysolidine along with two known spirobenzylisoquinoline alkaloids, isoochotensine and corysolidine. PMID- 11059824 TI - Hypoglycemic and hypolipidemic effects of tectorigenin and kaikasaponin III in the streptozotocin-lnduced diabetic rat and their antioxidant activity in vitro. AB - Tectorigenin and kaikasaponin III from the flowers of Pueraria thunbergiana showed potent hypoglycemic and hypolipidemic effects in the streptozotocin induced diabetic rats. Intraperitoneal administration of these two compounds with 5 and 10 mg/kg, respectively, for seven days to streptozotocin-induced rats significantly reduced the blood glucose, total cholesterol, LDL- and VLDL cholesterol and triglyceride levels when compared with those of control group. Glycitein in which 5-OH is unlinked and tectoridin (7-O-glycoside of tectorigenin) isolated from the flowers of P. thunbergiana did not improve hyperglycemia and hyperlipidemia. In addition, tectorigenin showed in vitro antioxidant effects on 1,1diphenyl-2-pirylhydrazyl (DPPH) radical, xanthine xanthine oxidase superoxide anion radical, and lipid peroxidation in rat microsomes induced by enzymatic and non-enzymatic methods. We further found that tectorigenin and kaikasaponin III protected the Vero cell line (normal monkey kidney) from injury by hydrogen peroxide. From these findings, it seems likely that the antioxidant action of tectorigenin and kaikasaponin III may alleviate the streptozotocin-induced toxicity and contribute to hypoglycemic and hypolipidemic effects. PMID- 11059825 TI - Furanocoumarins from the root of Angelica dahurica. AB - Five furanocoumarins including a new one were isolated from the root of Angelica dahurica by repeated silica gel column chromatography. Their chemical structures were determined to be isoimperatorin (1), oxypeucedanin hydrate-3"-butyl ether (2), imperatorin (3), knidilin (4), and oxypeucedanin hydrate (5). This represents the first study in which the compound 2 has been isolated and identified. The long-range coupling (5J) in the 1H-NMR spectrum observed in the linear furanocoumarin skeleton was also investigated in detail. PMID- 11059826 TI - Erectogenic effect of the selective phosphodiesterase type 5 inhibitor, DA-8159. AB - DA-8159, a new phosphodiesterase 5 inhibitor, was assessed for its erectogenic potential by a penile erection test in rats, the relaxation of isolated rabbit corpus cavernosum (CC), and estimation of the intracavernous pressure (ICP) in the anesthetized dog. Oral administration of DA-8159 (0.3 to 1 mg/kg) increased the number of erections in rats with increasing dosage, with the highest penile erection index at 10 mg/kg. DA-8159 induced the relaxation of phenylephrine (PHE) induced contractions in the rabbit CC and decreased the IC50 of the nitric oxide donor sodium nitroprusside (SNP) in a dose-dependent fashion. In pentobarbital anesthetized dogs, the intravenous administration of DA-8159 (1 approximately 300 g/kg) potentiated the increase in ICP induced by the intracavernosal SNP in a dose-related manner. These findings suggest that DA-8159 has significant therapeutic potential in the treatment of erectile dysfunction. PMID- 11059827 TI - Cytotoxicity and L-amino acid oxidase activity of crude insect drugs. AB - The cytotoxicity of crude insect drugs was measured using HeLa cells originating from human cervix and uterine cancer, using the dye uptake assay in order to find potential anticancer agents. Three kinds of extracts (buffer, methanol and ethylacetate) were prepared from 26 insects and used as raw materials for the activity assay. Among these, the buffer extracts from Tabanus, Mylabris and Huechys showed a potent anticancer activity, and those from Catharsius, Red ant, Scorpion, Tabanus and Vespae Nidus showed a strong L-amino acid oxidase (AAO) activity as well as cytotoxicity. In contrast, buffer extracts from Gryllotalpa orientalis and Apriona germari larvae showed greater/more rapid Hela cell growth than that of other insects. PMID- 11059828 TI - Antitumor activity of Bifidobacterium spp. isolated from a healthy Korean. AB - The antitumor activity of Bifidobacterium breve K-110, and K-111, and B. infantis K-525 was investigated. These Bifidobacterial cells and their cell wall preparations (WPG) significantly increased the survival rate of mice who had been intraperitoneally implanted with sarcoma 180 cells. Solid tumor growth was inhibited even when the sarcoma 180 cells were implanted into the groins of the mice. However, the Bifidobacterial cells did not show in vitro cytotoxicity against tumor cell lines. Cell kinetic studies revealed that these WPGs induced neutrophils, which were followed by macrophages, at the site of peritoneal injection. The WPGs directly activated these cells to inhibit the growth of tumor cells in in vitro assays. Our results suggest that Bifidobacterial WPGs induce and activate nonspecific phagocytes in situ to reject growing tumor cells in the mouse peritoneal cavity. PMID- 11059829 TI - Subacute nicotine exposure in cultured cerebellar cells increased the release and uptake of glutamate. AB - Cerebellar granule and glial cells prepared from 7 day-old rat pups were used to investigate the effects of sub-acute nicotine exposure on the glutamatergic nervous system. These cells were exposed to nicotine in various concentrations for 2 to 10 days in situ. Nicotine-exposure did not result in any changes in cerebellar granule and glial cell viability at concentrations of up to 500 microM. In cerebellar granule cells, the basal extracellular levels of glutamate, aspartate and glycine were enhanced in the nicotine-exposed granule cells. In addition, the responses of N-methyl-D-aspartate (NMDA)-induced glutamate release were enhanced at low NMDA concentrations in the nicotine-exposed granule cells. However, this decreased at higher NMDA concentrations. The glutaminase activity was increased after nicotine exposure. In cerebellar glial cells, glutamate uptake in the nicotine-exposed glial cells were either increased at low nicotine exposure levels or decreased at higher levels. The inhibition of glutamate uptake by L-trans-pyrollidine-2,4-dicarboxylic acid (PDC) was lower in glial cells exposed to 50 microM nicotine. Glutamine synthetase activity was lower in glial cells exposed to 100 or 500 microM of nicotine. These results indicate that the properties of cerebellar granule and glial cells may alter after subacute nicotine exposure. Furthermore, they suggest that nicotine exposure during development may modulate glutamatergic nervous activity. PMID- 11059830 TI - Effect of S-adenosylmethionine on hepatic injury from sequential cold and warm ischemia. AB - We investigated whether S-adenosylmethionine (SAM) treatment improved ischemic injury using perfused rat liver after sequential periods of 24 h cold and 20 min re-warming ischemia. SAM (100 micromol/L) was added to University of Wisconsin (UW) solution and Ringers lactate solution. After cold and sequential warm ischemia, releases of lactate dehydrogenase (LDH) and purine nucleoside phosphorylase (PNP) markedly increased during reperfusion. The increase in PNP was significantly reduced by SAM treatment. While the concentration of reduced glutathione (GSH) in ischemic livers significantly decreased, the concentration of glutathione disulfide (GSSG) increased. This decrease in GSH and increase in GSSG were suppressed by SAM treatment. Lipid peroxidation was elevated in cold and warm ischemic and reperfused livers, but this elevation was also prevented by SAM treatment. Hepatic ATP levels were decreased in the ischemic and reperfused livers to 42% of the control levels. However, treatment with SAM resulted in significantly higher ATP levels and preserved the concentration of AMP in ischemic livers. Our findings suggest that SAM prevents oxidative stress and lipid peroxidation and helps preserve hepatic energy metabolism. PMID- 11059831 TI - Antifibrotic effect of Stephania tetrandra on experimental liver fibrosis induced by bile duct ligation and scission in rats. AB - We examined the antifibrotic effect of a methanol extract from Stephania tetrandra (ST) on experimental liver fibrosis. Liver fibrosis was induced by bile duct ligation and scission (BDL/S) in rats. In BDL/S rats, activity levels of aspartate transaminase (AST), alanine transaminse (ALT), alkaline phosphatase (ALP), concentration of total bilirubin in serum, and hydroxyproline content of the liver were significantly increased. The ST treatment (either 100 mg/kg/day or 200 mg/kg/day, p.o. for 4 weeks) in BDL/S rats reduced the serum AST, ALT and ALP activity levels significantly (p< 0.01). Similarly, when compared to the control group, the concentration of hydroxyproline in the livers of the BDL/S rats treated with 100mg or 200mg ST treated rats decreased by 40% and 33% respectively, when compared to the BDL/S control group (p<0.01). The morphological characteristics of fibrotic liver that were observed in the BDL/S control group, improved in the ST treated BDL/S group. In the fibrotic liver of BDL/S rats treated with ST, a marked reduction in the numbers of alpha smooth muscle cell actin positive stellate cells was observed. These results indicate that doses of either 100 or 200 mg/kg/day of methanol extract from S. tetrandra, had an antifibrotic effect in rats with liver fibrosis induced by bile duct ligation and scission. PMID- 11059832 TI - Optimization study on the formulation of roxithromycin dispersible tablet using experimental design. AB - This study set out to improve the physical and pharmaceutical characteristics of the present formulation using an appropriate experimental design. The work described here concerns the formulation of the dispersible tablet applying direct compression method containing roxithromycin in the form of coated granules. In this study 2(3) factorial design was used as screening test model and Central Composite Design (CCC) associated with response surface methodology was used as optimization study model to develop and to optimize the proper formulation of roxithromycin dispersible tablet. The three independent variables investigated were functional excipients like binder (X1), disintegrant (X2) and lubricant (X3). The effects of these variables were investigated on the following responses: hardness (Y1), friability (Y2) and disintegration time (Y3) of tablet. Three replicates at the center levels of the each design were used to independently calculate the experimental error and to detect any curvature in the response surface. This enabled the best formulations to be selected objectively. The effect order of each term to all response variable was X3> X2> X1> X1*X2> X2*X2> X2*X3> X3*X3> X1*X3> X1*X1 and model equations on each response variables were generated. Optimized compositions of formula were accordingly computed using those model equations and confirmed by following demonstration study. As a result, this study has demonstrated the efficiency and effectiveness of using a systematic formulation optimization process to develop the tablet formulation of roxithromycin dispersible tablet with limited experiment. PMID- 11059833 TI - Enterohepatic recycling of estrogen and its relevance with female fertility. AB - Enterohepatic recycling of estrogen after oral administration of 1 mg non radioactive estriol was studied in fourteen women selected as the control subjects and ten infertile women in whom the infertility was appearing to be of endocrine origin. The extent of enterohepatic recycling of estriol (E3) during the early follicular phase of menstrual cycle was assessed by monitoring during 48 h the urinary excretion of its two major metabolites i.e; estriol 16 alpha glucuronide (E3-16 alpha-CG) and estriol-3 glucuronide (E3-3-G). The change in urinary level of E3-3-G with respect to E3-16 alpha-G was considered to reflect the extent of enterohepatic recycling of estriol. Lower values of urinary output of both metabolites in the infertile women as compared with the control subjects and the urinary excretion profile of both metabolites during 48 h after estriol ingestion reveal that the reduced extent of enterohepatic recycling could possibly be one of the factors which contribute towards the incidence of infertility in women. PMID- 11059834 TI - Differential expression of protein kinase C subtypes during ginsenoside Rh2 lnduced apoptosis in SK-N-BE(2) and C6Bu-1 cells. AB - We examined the modulation of protein kinase C (PKC) subtypes during apoptosis induced by ginsenoside Rh2 (G-Rh2) in human neuroblastoma SK-N-BE(2) and rat glioma C6Bu-1 cells. Apoptosis induced by G-Rh2 in both cell lines was confirmed, as indicated by DNA fragmentation and in situ strand breaks, and characteristic morphological changes. During apoptosis induced by G-Rh2 in SK-N-BE(2) cells, PKC subtypes alpha, beta and gamma were progressively increased with prolonged treatment, whereas PKC delta increased transiently at 3 and 6 h and PKC epsilon was gradually down-regulated after 6 h following the treatment. On the other hand, PKC subtype zeta markedly increased at 24 h when maximal apoptosis was achieved. In C6Bu-1 cells, no significant changes in PKC subtypes alpha, gamma, delta, epsilon and zeta were observed during apoptosis induced by G-Rh2. These results suggest the evidence for a possible role of PKC subtype in apoptosis induced by G-Rh2 in SK-N-BE(2) cells but not in C6Bu-1 cells, and raise the possibility that G-Rh2 may induce apoptosis via different pathways interacting with or without PKC in different cell types. PMID- 11059835 TI - Developmental patterns of mST3GalV mRNA expression in the mouse: in situ hybridization using DIG-labeled RNA probes. AB - mST3GalV synthesizes ganglioside GM3, the precursor for simple and complex a- and b- series gangliosides, and the expression and regulation of mST3GalV (CMP-NeuAc: lactosylceramide alpha2,3-sialyltransferase) activity is central to the production of almost all gangliosides, a class of glycosphingolipids implicated in variety of cellular processes such as transmembrane signaling, synaptic transmission, specialized membrane domain formation and cell-cell interactions. To understand the developmental expression of mST3GalV in mice, we investigated the spatial and temporal expression of mST3GalV mRNA during the mouse embryogenesis [embryonic (E) days; E9, E11, E13, E15] by in situ hybridization with digoxigenin-labeled RNA probes. All tissues from E9 and E11 were positive for mST3GalV mRNA. On E13, mST3GalV mRNA was expressed in various neural and non neural tissues. In contrast to these, on E15, the telencephalon and liver produced a strong expression of mST3Gal V which was a quite similar to that of E13. In this stage, mST3GalV mRNA was also expressed in some non-neural tissues. These data indicate that mST3GalV is differently expressed at developmental stages of embryo, and this may be importantly related with regulation of organogenesis in mice. PMID- 11059836 TI - Regulation of cytokine production by exogenous nitric oxide in murine splenocyte and peritoneal macrophage. AB - Nitric oxide (NO), products of activated macrophages, have a great impact on the regulation of cytokine production. The role of NO in non-specific host cells is commonly accepted. On the contrary, its role as an immuno-regulatory molecule is still controversial. In this study, we have investigated the effect of NO on the production of cytokines from murine splenocytes and macrophages. S-nitroso-L glutathione inhibited the release of both interferone-gamma and interleukin-2 produced by Th1 cells and tumor necrosis factor-alpha and interleukin-1beta produced by macrophages, but did not affect the release of interleukin-4 and interleukin-10 produced by Th2 cells. These results suggest that NO exerts a down regulatory effect on the secretion of cytokines from Th1 cells and macrophages which are implicated in immune response. Thus, NO may have an important role as an immuno-modulatory as well as effector molecule in the immune system. PMID- 11059837 TI - Environmental tobacco smoke uptake by clothing fabrics. AB - The uptake of cigarette smoke by a variety of clothing fabrics was determined. The tested fabrics included wool, linen, cotton, silk, rayon, acetate and polyester. While a variety of factors effect such absorption, polyester fabrics were found to significantly take up less smoke than all the other tested cloths. A simple and rapid technique is described for measuring such absorption. PMID- 11059838 TI - Characterisation and source identification of PM10 aerosol samples collected with a high volume cascade impactor in Brisbane (Australia). AB - PM10 (particulate matter with aerodynamic diameter < 10 microm) samples of Brisbane air were collected and fractionated into six size fractions (< 0.5, 0.5 0.61, 0.61-1.3, 1.3-2.7, 2.7-4.9 and 4.9-10 microm) with a high volume cascade impactor. The chemical composition of the samples was analysed by techniques including Ion Beam Analysis. On average, 42% of the aerosol mass is in the > 2.7 microm size fraction, with the < 0.5-microm size fraction also contributes 41% of the aerosol mass. The composition of the < 1.3-microm aerosols is significantly different to that of the > 1.3-microm aerosols. The aerosol mass and concentrations of chemical components related to human activities show a bimodal size-distribution pattern, with most of the mass in the accumulation range (< 0.65 microm). The size geometric mean of aerosol mass is 0.96 microm in the samples collected from an industrial/residential site, and is 1.74 microm in the samples collected from a suburban site. The size geometric mean of concentrations of chemical components related to human activities ranges from 0.16 to 0.57 microm. The concentrations of crustal matter and sea salt show a unimodal size distribution pattern, and with geometric means of 3.73 and 4.12 microm, respectively. Four source factors were resolved by multivariate analysis techniques for the size-fractionated aerosol samples, namely the soil, sea salt, organics and vehicular exhausts factors. The source fingerprints of the factors vary in the size ranges and have implications on the formation and dispersal processes of the particles. On average, the soil and sea salt factors contribute more than 80% of the aerosol mass in the > 2.7-microm fractions, while the organics and vehicular exhausts factors explain almost all the aerosol mass in the < 0.61-microm fractions. PMID- 11059839 TI - Application of double focusing sector field ICP-MS for multielemental characterization of human hair and nails. Part II. A study of the inhabitants of northern Sweden. AB - Double-focusing sector-field, inductively coupled plasma-mass spectrometry was used for the determination of 71 elements in scalp hair and fingernail samples from an urban population group living in the north-east of Sweden. Samples (n = 114 for hair and n = 96 for nails) were taken from subjects without known occupational exposure to metals. From these results, concentration ranges were calculated and compared with published intervals. Statistical analysis was used to elucidate differences according to sex, age and smoking habit. It was found that significant correlations exist between different elements in hair and nails, as well as between hair and nail concentrations for several elements. Strong positive correlation for Hg, Cd, Pb, Sb and Bi levels between these media confirms that both can be used for exposure assessment for these elements. Several examples on the use of distribution patterns for the rare-earth elements (REE) and of Pb isotope ratios for assessment of exposure are given. PMID- 11059840 TI - Methyl mercury bioaccumulation in long-finned eels, Anguilla dieffenbachii, from three rivers in Otago, New Zealand. AB - This research focuses on mercury (Hg) bioaccumulation in New Zealand long-tinned eels (Anguilla dieffenbachii) from the aquatic environment. Total Hg (HgT) and methyl mercury (MeHg) concentrations were determined in muscle tissue from eels living in three South Island rivers dominated respectively by urban, native bush and agricultural land-uses. Most of the Hg in eels was MeHg (> 84%) and the MeHg concentrations increased linearly with both length and eel age for a given river habitat. The annual growth rates for eels from the urban and agricultural streams were greater than for eels from the native bush stream. The average MeHg accumulation rate was significantly higher for the eels in the agricultural stream compared with either the urban or native bush catchments. These results are probably due to a combination of factors and further investigations in the lower food web are necessary to elucidate the exact mechanisms of MeHg bioaccumulation in these creatures. PMID- 11059841 TI - Urinary excretion of lead during pregnancy and postpartum. AB - We have compared lead isotopic ratios and lead concentrations in 53 spot urine and 59 24-h urine samples from 13 subjects covering the interval from pre pregnancy through 180 days postpartum to estimate the amount of lead excreted in urine and renal clearance relative to blood. The total amount of lead excreted in 24-h urine samples ranges from 0.8 to 5.9 microg Pb with an arithmetic mean of 2.2+/-1.1 microg (geometric mean 1.90 microg). This compares with amounts of 0.9 10 microg of extra lead per day estimated to be released into blood from the skeleton during pregnancy and postpartum. There were no differences in excretion rates during the trimesters of pregnancy and between pregnancy and postpartum time periods. The renal clearance relative to blood ranged from 0.8 to 10 g/h (arithmetic mean 3.2+/-1.9; geometric mean 2.7). Renal clearance relative to blood was somewhat higher in trimesters 2 and 3 compared with postpartum 150-180 days (P = 0.004, 0.006, respectively). Reassessment of earlier published blood and dietary data for Australian pregnant controls indicates there is no increased gastrointestinal absorption of lead during pregnancy and postpartum. This differs from calcium, which shows increased absorption during late pregnancy. In light of the inconvenience of sampling and potential contamination at the low levels of lead found in most of these subjects, we do not consider the 24 h urines to provide sufficient useful information. PMID- 11059842 TI - An improved analytical procedure for determination of total actual acidity (TAA) in acid sulfate soils AB - An improved analytical procedure is proposed for the determination of total actual acidity (TAA) in acid sulfate soils. The proposed method involves the use of a superior extracting solution, 0.5 M BaCl2, instead of the 1 M NaCl used by Konsten et al. (Konsten CJM, Brinkman R, Andriesse W. A field laboratory method to determine total potential and actual acidity in acid sulfate soils. In: Dost H, editor. Selected papers of the Dakar Symposium on Acid Sulfate Soils. Wageningen: ILRI Publication 44, 1988:106-134.) and improved experimental design to obtain correction factors for calculating TAA. The introduction of a multi choice procedure also enables increased accuracy of analytical results to be obtained if more accurate TAA estimation is required. PMID- 11059843 TI - Ambient sulfate concentration and chronic disease mortality in Beijing. AB - In this study, ecological analysis was used to assess the relationship between ambient air pollution and human mortality. All the data on environmental measures and related factors, population size and number of deaths were collected for the city of Beijing, PR China and its eight districts for the years 1980-1992. In this study the concentration of SO(4)2- was selected as a main indicator of environmental pollution for the following reasons: (i) SO(4)2- data are available to cover all urban and suburban areas in Beijing compared with other air pollutants during the study period; (ii) SO(4)2- levels indicate the concentration of sulfide (include sulfate) and acid fog in the air, and they are significantly lower in cleaner districts than in others; and (iii) analyses showed that SO(4)2- levels are significantly correlated with daily mean concentrations of sulfur dioxide and nitrogen oxide, annual coal combustion, number of households using gas fuel, counts of motor vehicles and population density. Age-standardised mortality rates due to specific diseases were calculated using the Chinese population census data in 1990. Statistically significant correlations were observed between SO(4)2- concentration and total mortality and mortality due to cardiovascular disease, malignant tumour and lung cancer (r > 0.50 in all cases). The correlations were not only found between the current SO(4)2- concentration and these mortalities, but also for SO(4)2- levels measured up to 12 years prior to death, which may suggest long-term effects of air pollution. No significant correlations were observed for mortality from respiratory diseases and cerebrovascular diseases (r = 0.30-0.50). This study indicates that the concentration of SO(4)2- in air is a useful air pollution indicator in the areas where coal is used as the main source of energy. Areas with high levels of SO(4)2- experienced higher mortality due to a variety of chronic diseases. PMID- 11059844 TI - 137Cs and relationships with major and trace elements in edible mushrooms from Mexico. AB - 137Cs and 40K specific activity together with major and trace elements were determined in soil samples and in different edible wild mushroom species collected from a seminatural temperate forest ecosystem located in the central part of the Mexican Volcanic Belt. The activity measurements were made using a gamma-ray spectrometer system with a high purity germanium (HpGe) detector. The major and trace elements were determined using emission spectrography and mass spectrometry, respectively. The aggregated transfer factors for 137Cs were estimated in 30 local mushroom species collected from 1993 to 1999. Differences as large as three orders of magnitude were observed. The contribution of mushrooms for the total 137Cs dietary intake by the local population was estimated to be 37%. Mushrooms also showed to be good accumulators for Rb, Cu, Cs and Se. PMID- 11059845 TI - Estimation of apparent rate coefficients for radionuclides interacting with marine sediments from Novaya Zemlya. AB - To assess the impact of radionuclides entering the marine environment from dumped nuclear waste, information on the physico-chemical forms of radionuclides and their mobility in seawater-sediment systems is essential. Due to interactions with sediment components, sediments may act as a sink, reducing the mobility of radionuclides in seawater. Due to remobilisation, however, contaminated sediments may also act as a potential source of radionuclides to the water phase. In the present work, time-dependent interactions of low molecular mass (LMM, i.e. species < 10 kDa) radionuclides with sediments from the Stepovogo Fjord, Novaya Zemlya and their influence on the distribution coefficients (Kd values) have been studied in tracer experiments using 109Cd2+ and 60Co2+ as gamma tracers. Sorption of the LMM tracers occurred rapidly and the estimated equilibrium Kd(eq)-values for 109Cd and 60Co were 500 and 20000 ml/g, respectively. Remobilisation of 109Cd and 60Co from contaminated sediment fractions as a function of contact time was studied using sequential extraction procedures. Due to redistribution, the reversibly bound fraction of the gamma tracers decreased with time, while the irreversibly (or slowly reversibly) associated fraction of the gamma tracers increased. Two different three-compartment models, one consecutive and one parallel, were applied to describe the time-dependent interaction of the LMM tracers with operationally defined reversible and irreversible (or slowly reversible) sediment fractions. The interactions between these fractions were described using first order differential equations. By fitting the models to the experimental data, apparent rate constants were obtained using numerical optimisation software. The model optimisations showed that the interactions of LMM 60Co were well described by the consecutive model, while the parallel model was more suitable to describe the interactions of LMM 109Cd with the sediments, when the squared sum of residuals were compared. The rate of sorption of the irreversibly (or slowly reversibly) associated fraction was greater than the rate of desorption of the reversibly bound fractions (i.e. k3 > k2) for both radionuclides. Thus, the Novaya Zemlya sediment are supposed to act as a sink for the radionuclides under oxic conditions, and transport to the water phase should mainly be attributed to resuspended particles. PMID- 11059846 TI - Regional ecological risk assessment of selenium in Jilin province, China. AB - Integrating the biokinetic model of selenium with Monte Carlo analysis, this article carries out a quantitative study on ecological risk assessment in blood selenium levels of residents in Jilin province. The result shows that the established biokinetic model can be employed to predict the blood selenium levels of residents in a region. The predicted average blood selenium is 0.044 microg/ml and standard deviation is 0.013 microg/ml. Compared with the determined average blood selenium (0.043 microg/ml) and standard deviation (0.019 microg/ml), there was no obvious difference between each other (P > 0.05). The predictive risk degree (19.8%) is also similar to that of the Keshan disease occurrence (16.4%) of residents in Jilin province. The advance of the ecological risk assessment model of selenium offers a good example for the study on predictive models of other trace elements in the human body. PMID- 11059847 TI - Heavy metal accumulation in the periwinkle Littorina littorea, along a pollution gradient in the Scheldt estuary. AB - Inductively coupled plasma atomic emission spectrophotometry (ICP-AES) was used to determine heavy metal concentrations in the shells and soft tissues of the periwinkle Littorina littorea, collected at seven sites along the Scheldt estuary. Several metals accumulate in the animals soft body parts, and are related to the seawards decreasing pollution gradient. No clear correlation with a previously detected shell size patterning could be established. PMID- 11059848 TI - The effect of large anthropogenic particulate emissions on atmospheric aerosols, deposition and bioindicators in the eastern Gulf of Finland region. AB - The effect of the emissions from large oil shale fuelled power plants and a cement factory in Estonia on the elemental concentration of atmospheric aerosols, deposition, elemental composition of mosses and ecological effects on mosses, lichens and pine trees in the eastern Gulf of Finland region has been studied. In addition to chemical analysis, fly ash, moss and aerosol samples were analysed by a scanning electron microscope with an energy dispersive X-ray spectrometer (SEM/EDS). The massive particulate calcium emissions, approximately 60 kton/year (1992), is clearly observed in the aerosols, deposition and mosses. The calcium deposition is largest next to the Russian border downwind from the power plants and in south-eastern part of Finland. This deposition has decreased due to the application of dust removal systems at the particulate emission sources. At the Virolahti EMEP station approximately 140 km north from the emission sources, elevated elemental atmospheric aerosol concentrations are observed for Al, Ca, Fe, K and Si and during episodes many trace elements, such as As, Br, Mo, Ni, Pb and V. The acidification of the soil is negligible because of the high content of basic cations in the deposition. Visible symptoms on pine trees are negligible. However, in moss samples close to the power plants, up to 25% of the leaf surface was covered by particles. Many epiphytic lichen species do not tolerate basic stemflow and on the other hand most species are also very sensitive for the SO2 content in air. Consequently a large lichen desert is found in an area of 2500 km2 in the vicinity of the power plants with only one out of the investigated 12 species growing. PMID- 11059849 TI - Changes in rainfall chemistry and airborne particulates during a period of major local industrial change. AB - Data from a site in central Scotland were used to quantify the changes in rainfall quality from 1989 to 1998. During this period there have been major changes in industrial activities in the area, particularly the decline in local steel-making and steel-processing activities. Many element concentrations in rainfall decreased over time in parallel with the phased reduction in the activity of local pollutant sources. Trend analyses of the rainfall data identified that the most significant responses have been the lower concentrations of Ca, SO4-S and Mn. There was also a dramatic decline in the capture of airborne particulates by the interception rainfall gauges. Particulates were found to contain mainly hematite, magnetite and quartz, that is similar to what would be expected to be derived from the neighbouring steel industries. The eventual disappearance of these particulates and the responses in rainfall quality match the timescale for the decline and closure of some of the potential sources of pollutants. PMID- 11059850 TI - Effects of oral exposure to polychlorinated biphenyls (PCBs) on the development and metamorphosis of two amphibian species (Xenopus laevis and Rana temporaria). AB - This study examined the effects of polychlorinated biphenyls (PCBs) on development of families of amphibians using the African clawed frog (Xenopus laevis) and the European common frog (Rana temporaria). Amphibians were orally exposed to the technical PCB-mixture Clophen A50 or to the non-ortho-3,3',4,4',5 CB congener (PCB 126) either for a 10-day period or until metamorphosis. Occurrence and rate of malformations, mortality, period until metamorphosis and thyroid hormone levels were measured. Mortality increased in a dose-dependent manner, as did the rates of malformation. Time until metamorphic transformation was prolonged and the weight of froglets was increased. Although not statistically significant, thyroid hormone levels were also lowered. PHAHs such as PCBs may affect important aspects of amphibian fitness and may influence amphibian reproductive success. PMID- 11059851 TI - The application of reporter gene assays for the determination of the toxic potency of diffuse air pollution. AB - Diffuse air pollution consists of a mixture of numerous compounds. It is emitted by many distributed sources and is omnipresent due to atmospheric transport. Risk assessment of the complex mixture of air pollutants on the basis of the toxicity of the individual compounds is not yet possible because the chemical identity and/or toxicity of the constituencies of a substantial fraction is unknown. In addition, no adequate procedures are available to integrate toxicity data of such complex mixtures, so that an individual risk assessment of the constituents of air pollution disregards possible combination effects. In the present study, an approach has been developed to assess the toxic potency by using in vitro bio assay techniques. Genotoxicity was assessed in the umu-assay, a reporter gene assay using a strain of Salmonella typhimurium stably transfected with a plasmid (pSK1002) carrying the SOS-gene umuC fused to the reporter gene lacZ. Arylhydrocarbon-receptor activation was assessed in the DR-CALUX-assay, using a stably transfected H4IIE hepatoma cell line containing a plasmid for the luciferase gene under transcriptional control of dioxin-responsive elements. Samples of airborne particulate matter (APM) were collected with a high volume sampler next to a highway and in a natural conservation area. Both assays proved to be applicable to quantify genotoxicity and the presence of polycyclic aromatic hydrocarbons (PAHs) in small extracts from air-filter samples. Results indicate that PAHs from traffic exhausts seem to be largely responsible for an increased genotoxic activity of APM collected down-wind from the highway (western wind). APM collected at eastern wind directions seems to have a different composition of compounds, with a higher genotoxic activity that is less related to highway emitted PAH-like compounds. At northern wind directions, APM is relatively less genotoxic and contains less PAHs than at other wind directions. Dioxin-like compounds contribute negligibly to the Ah-receptor agonistic potency of APM. Airborne pollutants with genotoxic and/or PAH-like characteristics form an undesired mutagenic risk, which will be evaluated in further in vivo studies. PMID- 11059852 TI - Heavy metals in rivers of Latvia. AB - Total heavy metal concentrations in waters and sediments (HNO3 digestible Pb, Cu, Co, Ni, Mn, Zn) and their speciation forms in sediments (exchangeable, carbonate bound, iron-manganese oxide bound, organic matter bound and residual) in major and common small watercourses (31 sampling stations) along their flow in Latvia were determined. The metal loads entering the Baltic Sea from Latvia were calculated. Increased metal concentrations were found only in lower reaches of the largest rivers and locally around known industrial pollution sources. Differences in metal concentrations and loads in rivers from different regions of Latvia were related to natural geochemical processes. Metal speciation analysis showed that the dominant metal species are residual metals and those bound to organic matter. Residual and carbonate-bound metal dominated only in rhitral regions of rivers. The concentrations of exchangeable metals increased below pollution sources. PMID- 11059853 TI - Neuro-endocrine biomarkers of pollution-induced stress in marine invertebrates. AB - Nitric oxide (NO) is a molecular mediator of the non-specific immune response of vertebrates but production has also been recorded in response to bacterial infection in invertebrates. Together with biogenic amines and cytokines, production of NO in invertebrate phagocytic haemocytes is thought to represent a putative stress response. In this study the hypothesis that NO might be a sensitive biomarker of stress in invertebrates has been tested. The spectrophotometric Griess assay for nitrite and nitrate has been modified so that the final breakdown products of NO are suitable for measurement in invertebrate haemolymph. The assay was linear in the range 1-100 microM with a sensitivity of 4 microM. The baseline NOx measurement in unstressed mussels was 0.74 mM NOx/mg protein. Mytilus edulis were kept at 15 micro C in filtered seawater for at least 2 days prior to exposure to TBTO (0.001-0.1 mg/l) for 24 h. Preliminary results suggest that TBTO perturbs the NO response and that the outlined assay protocol is a sensitive means of detecting those changes. It is proposed that NO measurements potentially offer a highly sensitive, non-invasive means of monitoring stress responses associated with environmental change. PMID- 11059854 TI - Reporting personal results to participants of exposure studies. AB - In most exposure studies personal results are reported to the participating subjects. This paper describes how this was achieved in two different studies, the large scale German Environmental Survey and the smaller Helsinki part of the multi-national, multi-centre EXPOLIS study. In spite of the different approaches both independently reported personal results in a very similar fashion, involving automation and graphical display of measured values. PMID- 11059855 TI - Magnetic resonance cholangiography in patients with biliary disease: its role in primary sclerosing cholangitis. AB - BACKGROUND/AIM: Magnetic resonance cholangiography (MRC) is a non-invasive diagnostic procedure whose role in the management of patients with primary sclerosing cholangitis (PSC) is unclear. The aim of this study was to determine the usefulness of MRC in the evaluation of the biliary tree in patients with suspected biliary disease, and in particular, PSC. METHODS: MRC and invasive cholangiography (ERCP or PTC) were both performed in 73 patients, (33 male, 40 female, mean age 56 years) with clinical and/or biochemical evidence of cholestasis. Images were interpreted by two radiologists unaware of the results of other studies. RESULTS: Forty-two patients (58%) had benign biliary disease, including 23 patients (32%) with PSC; 9 patients (12%) had malignant biliary disease; and 22 patients (30%) had a normal biliary tree. Diagnostic quality images were obtained in 73/73 (100%) of MRC, and in 70/73 (96%) of invasive cholangiography (68 ERCP's, 2 PTC's) procedures. Using ERCP/PTC findings as the reference standard, MRC had an accuracy greater than 90% in the diagnosis of normal bile ducts, biliary dilatation, biliary obstruction, bile duct stones, and PSC. Using the final diagnosis, MRC had an overall diagnostic accuracy of 90% in the detection of biliary disease compared to 97% for invasive cholangiography. Additional diagnostic/therapeutic interventions were performed during ERCP in 73% of patients with PSC and in 43% of patients without PSC (p=0.02). CONCLUSIONS: MRC has excellent diagnostic accuracy in the presence of biliary disease. Because of its noninvasive nature, MRC may have advantages over invasive cholangiography when diagnosis is the major goal of the procedure. PMID- 11059856 TI - Antibodies against the COOH-terminal region of E. coli ClpP protease in patients with primary biliary cirrhosis. AB - BACKGROUND/AIMS: The presence of antibodies in sera from patients with autoimmune diseases is an important tool for diagnosis and for providing insights into the mechanisms leading to autoimmunity. The aim of this study was to characterize new reactive antigens in liver autoimmune diseases. METHODS: Sera of patients with liver-related autoimmune (n=74) and non-liver-related autoimmune (n= 211) diseases, non-autoimmune liver diseases (n=18) and healthy controls (n=160) were evaluated for antibodies against E. coli ClpP protease (EClpP) and 20S proteasome by immunoblot analysis. RESULTS: Antibodies against EClpP were detected in 15 of 50 patients with primary biliary cirrhosis, in only one of 100 patients with systemic lupus erythematosus, and in three healthy subjects (Chi-square 59.1, d.f. 2, p< 0.001). Antibodies to 20S proteasome were found in only 35 of 100 patients with systemic lupus erythematosus. All other sera from patients with autoimmune diseases, liver diseases other than primary biliary cirrhosis, and healthy controls were negative for both antigens. Both IgG and IgM classes of antibodies against EClpP were present in primary biliary cirrhosis patient sera with titers of 1/400-1/1000. By using recombinant techniques and peptide ELISA, the immunodominant EClpP epitope recognized by the sera from primary biliary cirrhosis patients was localized in the amino acid sequences 177-194 (QIERDTERDRFLSAPEAV) within the COOH-terminal of EClpP. Affinity-purification of these anti-EClpP antibodies and immunoabsorption experiments established that the antibodies are specific for the bacterial EClpP. CONCLUSIONS: Bacterial ECIpP has been identified as a new antigen specifically reacting with sera from approximately one third of patients with primary biliary cirrhosis. PMID- 11059857 TI - Overlap of autoimmune hepatitis and primary sclerosing cholangitis: an evaluation of a modified scoring system. AB - BACKGROUND/AIM: Recently, the scoring system for the diagnosis of autoimmune hepatitis (AIH) was modified by the International AIH Group. Our aim was to determine the prevalence of AIH in patients with cholangiographically proven primary sclerosing cholangitis (PSC) using this new scoring system. METHODS: A total of 211 PSC patients were evaluated. RESULTS: Three (1.4%) patients scored more than 15 points ('definite' AIH); 13 (6%) patients scored between 10 and 15 points ('probable' AIH); the remaining 195 (93%) patients had less than 10 points, allowing the exclusion of AIH. The separation of patients with PSC plus AIH from patients with PSC alone was based mostly on serum levels of total globulins (p=0.01), IgG (p=0.001), titers of autoantibodies (p<0.001) and histologic score (p<0.001). Using the older scoring system, four (2%) patients met the criteria for the diagnosis of PSC plus 'definite' AIH and 40 (19%) the diagnosis of PSC plus 'probable' AIH. CONCLUSIONS: Overlap of PSC and AIH occurs rarely. The new scoring system seems to more precisely define the potential overlap syndrome between PSC and AIH, although further modification of the new scoring system may provide even better discrimination among these conditions. PMID- 11059858 TI - High prevalence of autoimmune hepatitis among patients with primary sclerosing cholangitis. AB - BACKGROUND/AIMS: Traditionally, autoimmune hepatitis (AIH) and primary sclerosing cholangitis (PSC) are regarded as separate disease entities. We report on a group of patients that suggests the existence of an overlap syndrome of the two conditions and on the prevalence of this syndrome among patients with PSC. Furthermore, the impact of the recently revised AIH scoring system for diagnosing AIH in this context was assessed. METHODS: Retrospective analysis of consecutive patients of a tertiary referral centre for liver disease with a diagnosis of PSC. RESULTS: Diagnosis of the overlap syndrome was established for nine patients (8%) of a total group of 113 PSC patients. Four patients initially presented with features of AIH and in five cases PSC was diagnosed first. All patients responded to immunosuppressive therapy; in three cases long-term remission was achieved. Three patients underwent liver transplantation after 4 months and 7 and 9 years, respectively. The original and revised versions of the AIH scoring system gave essentially the same results in the patients with the PSC-AIH overlap syndrome. CONCLUSIONS: Patients with overlapping features of AIH and PSC may be more common than is currently assumed. Recognition of this syndrome is of clinical significance, considering the important therapeutical consequences. PMID- 11059859 TI - Effects of budesonide and prednisolone on hepatic kinetics for urea synthesis. AB - BACKGROUND/AIMS: Glucocorticoids upregulate hepatic urea synthesis and cause protein breakdown to prevail over synthesis, releasing amino acids into the blood stream and increasing the substrate supply for hepatic urea synthesis. Budesonide is a new generation glucocorticoid that may be used for treatment of inflammatory diseases, e.g. Crohn's disease and autoimmune hepatitis. Due to its extensive first-pass metabolism in the liver, it has a potential adverse effect profile superior to that of prednisolone. Little attention has been directed towards differences in nitrogen catabolic properties between budesonide and prednisolone. METHODS: Eight normal male subjects (age 20-44 years; BMI 21.6-28.2 kg/m2) were randomly studied 3 times: 1) At baseline, 2) after 6 days of prednisolone (50 mg/day), and 3) after 6 days of budesonide (9 mg/ day). We measured urea nitrogen synthesis rates (UNSR) and blood alpha-amino-nitrogen (N) levels before, during, and after a 3-h constant infusion of alanine (2 mmol/(kg BW x h)). UNSR was estimated hourly as urinary excretion corrected for gut hydrolysis and accumulation in body water. The slope of the linear relationship between UNSR and amino-N concentration represents the hepatic kinetics of conversion of amino- to urea-N, and is denoted the functional hepatic nitrogen clearance (FHNC). RESULTS: Prenisolone, but not budesonide, administration increased basal blood and amino nitrogen concentrations (3.5 +/- 0.1 mmol/l (control) vs 3.8 +/- 0.1 mmol/l (prednisolone) (p<0.05) and 3.6 +/- 0.1 mmol/l (budesonide) (NS). Basal UNSR values were significantly increased following prednisolone (23.3 +/- 6.5 (control) vs 51.2 +/- 6.3 (prednisolone) (p<0.05)), while budesonide had no effect on basal UNSR (33.7 +/- 4.2 (budesonide) (NS)). Prednisolone administration increased FHNC (from 24.6 +/- 4.7 l/h (control) to 47.3 +/- 5.9 l/h (prednisolone) (p<0.05). Budesonide administration did not significantly increase FHNC (33.7 +/- 4.2 l/h (budesonide), (vs control; p=0.12, vs prednisolone: p<0.05)). CONCLUSIONS: Prednisolone administration led to increased levels of amino acids in blood and loss of N as urea, the latter in part due to a specific hepatic mechanism as shown by the increased FHNC. Budesonide led to unaltered levels of amino acids in blood, no changes in loss of N as urea, and unaltered hepatic kinetics for urea synthesis. Thus, oral budesonide administration had very limited effects on the hepatic contribution to nitrogen homeostasis and metabolism via urea synthesis, making treatment with budesonide superior to that of conventional glucocorticoids in this respect. PMID- 11059860 TI - Hepatic microvascular features in experimental cirrhosis: a structural and morphometrical study in CCl4-treated rats. AB - BACKGROUND/AIMS: In this study, a detailed morphometrical analysis of the hepatic microvasculature in the different zones of hepatic parenchyma was performed in normal and cirrhotic rat liver (CCl4-induced). The aims were to detect, in CCl4 induced cirrhosis, the real presence of the "capillarization" of hepatic sinusoids and to assess alterations of the sinusoid/parenchyma ratio within the nodule. METHODS: Cirrhosis was promoted by controlled intragastric CCl4 administration. Scanning electron microscopy of the vascular corrosion cast technique associated with light microscopy and transmission electron microscopy were used. RESULTS: Evidence of connective tissue in the space of Disse was found only in sinusoids located near portal tracts or large fibrotic areas, and this was also confirmed by laminin immunohistochemistry. In contrast, all the intranodular sinusoids lacked real basal membrane and connective fibers in the space of Disse and, displayed normal fenestrations. The parenchymal area, sinusoidal area, mean sinusoidal area, sinusoidal perimeter, hepatocyte area and the reciprocal ratios were all considered in the morphometrical analysis. The sinusoids were of uniform size in the periportal, periseptal and pericentral areas of the cirrhotic liver without the typical zonal differences of the normal liver. The areas occupied by sinusoids per unit of parenchyma and the sinusoid/hepatocyte interfaces disposable for metabolic exchanges were markedly smaller (p<0.01) in cirrhotic than normal liver. CONCLUSION: Our findings indicate that capillarization of hepatic sinusoids occurs only in very limited regions of the cirrhotic parenchyma, and thus this phenomenon does not have relevant functional consequences. Furthermore, the cirrhotic parenchyma appears not to be supplied by sinusoids and lacks features of zonation, which is a condition that could play a major role in the development and progression of liver failure. PMID- 11059861 TI - Oral ciprofloxacin after a short course of intravenous ciprofloxacin in the treatment of spontaneous bacterial peritonitis: results of a multicenter, randomized study. AB - BACKGROUND/AIMS: Oral quinolones have been suggested as treatment of cirrhotic patients with uncomplicated spontaneous bacterial peritonitis. To evaluate the efficacy of oral quinolones in all patients with this complication, oral ciprofloxacin after a short course of intravenous (i.v.) ciprofloxacin was compared to i.v. ciprofloxacin. METHODS: Eighty patients were allocated to receive ciprofloxacin i.v. 200 mg/12 h for 7 days (group A, n= 40) or i.v. 200 mg/12 h during 2 days followed by oral 500 mg/12 h for 5 days (group B, n=40). All patients with spontaneous bacterial peritonitis admitted to the hospital were included. Twenty-five variables obtained 48 h after treatment were introduced into univariate and multivariate analyses to identify predictors of survival and outcome. RESULTS: In the baseline condition, no differences were found between the two groups in clinical data, hepatic and renal function tests and Child Pugh score. The infection resolution rate was 76.3 % in group A and 78.4 % in group B, and hospital survival was 77.5% in both groups. In multivariate analysis serum creatinine and serum leukocytes 48 h after treatment were associated with prognosis. CONCLUSIONS: Oral ciprofloxacin after a short course of i.v. ciprofloxacin is effective in the treatment of spontaneous bacterial peritonitis. This regimen can be applied to all patients admitted to the hospital with this complication, and could be an alternative to treating these patients as outpatients. PMID- 11059862 TI - HSP70 induction by cyclosporine A in cultured rat hepatocytes: effect of vitamin E succinate. AB - BACKGROUND/AIMS: The effect of vitamin E succinate was studied in vitro against cyclosporine A (CsA) cytotoxicity in reference to the induction of heat shock protein 70 (HSP70) in rat hepatocytes. METHODS: Primary cultures of hepatocytes were incubated with CsA, in the range of 0 to 50 microM, in the presence or absence of 50 microM vitamin E succinate for 24 h. Peroxides were quantified by using 2',7'dichlorodihydrofluorescein diacetate, HSP70 levels were measured by Western blot analysis and apoptosis was detected by the hypodiploid peak of propidium iodide stained DNA. RESULTS: At 24 h of incubation with CsA, intracellular peroxide content increased in a dose-dependent manner. HSP70 also increased in parallel to CsA concentration, and apoptosis showed a biphasic change, increasing at concentrations between 0 and 10 microM and decreasing from 10 to 50 microM. The effect of vitamin E was studied at 24 h of coincubation with CsA. The values obtained show that vitamin E significantly counteracted the effect of CsA, diminishing the CsA-induced increase in intracellular peroxides and the lysis of cell membrane. These vitamin E effects were accompanied by a decrease in HSP70 and an enhancement of apoptosis. CONCLUSIONS: We conclude that in primary hepatocyte cultures, peroxide generation and cytotoxicity induced by CsA was accompanied by HSP70 induction and that the CsA cytotoxicity significantly decreased in the presence of vitamin E succinate parallel to a disappearance in HSP70 and to an increase in apoptosis. PMID- 11059863 TI - Hepadnaviral hepatocarcinogenesis: in situ visualization of viral antigens, cytoplasmic compartmentation, enzymic patterns, and cellular proliferation in preneoplastic hepatocellular lineages in woodchucks. AB - BACKGROUND/AIMS: Hepadnaviral hepatocarcinogenesis induced in woodchucks with and without dietary aflatoxin B1 has been established as an appropriate animal model for studying the pathogenesis of human hepatocellular carcinoma in high-risk areas. Our aim in this study was the elucidation of phenotypic cellular changes in early stages of this process. METHODS: Woodchucks were inoculated as newborns with woodchuck hepatitis virus (WHV), and partly also exposed to aflatoxin B1. Sequential hepatocellular changes in the expression of viral antigens, ultrastructural organization, cellular proliferation and apoptosis were studied in situ by electron microscopy, enzyme and immunohistochemistry. RESULTS: A characteristic finding in WHV-infected animals (with and without aflatoxin B1) was proliferative areas of minimal structural deviation, which predominated periportally, comprised glycogen-rich, amphophilic, and ground-glass hepatocytes, and expressed the woodchuck hepatitis core and surface antigens. Two main types of proliferative foci emerged from minimal deviation areas, glycogenotic clear cell foci and amphophilic cell foci (being poor in glycogen but rich in mitochondria), giving rise to the glycogenotic-basophilic and the amphophilic preneoplastic hepatocellular lineages. A gradual loss in the expression of viral antigens appeared in both lineages, particularly early in the glycogenotic basophilic cell lineage. Whereas glycogenosis was associated with an enzymic pattern suggesting an early activation of the insulin-signaling pathway, amphophilic cells showed changes in enzyme activities mimicking a response of the hepatocytes to thyroid hormone, which may also result from early changes in signal transduction. CONCLUSION: Preneoplastic hepatocellular lineages in hepadnaviral and chemical hepatocarcinognesis show striking phenotypic similarities, indicating concordant and possibly synergistic early changes in signaling. PMID- 11059864 TI - In vitro and in vivo suppression of growth of rat liver epithelial tumor cells by antisense oligonucleotide against protein kinase C-alpha. AB - BACKGROUND/AIMS: It has been hypothesized that liver stem cells may be activated and proliferate upon liver injury and may participate in the development of liver cancer. GP7TB, a rat liver epithelial tumor cell line, possesses characteristics of liver stem-like cells and can develop into a tumor in syngeneic Fischer 344 rat. We found that protein kinase C-alpha (PKC-alpha) is overexpressed in GP7TB cells. The importance of PKC-alpha for this liver tumor cell was elucidated. METHODS: Antisense oligonucleotide (ODN) was applied to suppress the production of PKC-alpha in GP7TB cells in vitro and in vivo. Cell viability was measured by acid phosphatase assay. The cellular levels of PKC-alpha and Bcl-2 were monitored by Western-blot analysis. Activation of nuclear factor NF-kappaB was analyzed by electrophoretic mobility shift assay. Cell cycle phase distribution was monitored by FACScan. Cell apoptosis was detected by TUNEL assay and histochemical staining of tumor tissue sections. The in vivo experiment was conducted by implanting tumor mass of GP7TB in the liver of F-344 rat and continuous delivery of the ODN by a mini-osmotic pump. RESULTS: Antisense ODN effectively suppressed the level of PKC-alpha that resulted in the decrease of Bcl-2 and nuclear NF-kappaB. The cumulative viable cells also decreased dramatically for the antisense-treated group. FACScan showed that the cells were arrested at early S-phase. These cells in turn went into apoptosis without completing a cell cycle. It was found that growth of the tumor was suppressed efficiently by antisense ODN. Cell apoptosis was found in the orthotopic tumor. The normal liver cells were not affected. CONCLUSIONS: A lethal effect of depressing the level of PKC-alpha in GP7TB cells and success in suppressing orthotopic tumor growth in vivo suggests that PKC alpha antisense ODN would be a promising therapeutic agent for some liver cancers. PMID- 11059865 TI - The impact of liver disease and medical complications on quality of life and psychological distress before and after liver transplantation. AB - BACKGROUND/AIM: The impact of liver disease and medical complications on quality of life (QOL) and psychological distress before and after orthotopic liver transplantation (OLT) is a matter of growing interest. METHODS: Perceived QOL (LEIPAD Quality of Life test) and psychological distress (Brief Symptom Inventory, BSI) were assessed in 40 cirrhotic patients listed for OLT (Group A) and in 101 liver transplant recipients (Groups B to G=0-6, 7-12, 13-24, 25-36, 37 48, 49-60 months post-OLT). Patients were also evaluated for medical complications, blood levels of immunosuppressive agents and recurrence of liver disease. RESULTS: QOL and psychological distress were significantly better in most of the post-OLT groups than in cirrhotic patients. Among post-OLT patients, a significantly worse QOL was perceived at 13-24 months (Life Satisfaction: Group D vs G, p=0.024; Cognitive Functioning: Group D vs F, p=0.024), while significantly greater psychological distress was detected at 7-12 months (Anxiety and Interpersonal Sensitivity: Group C vs Group B, p=0.032 and p=0.023, respectively). Medical complications and immunosuppressive therapy did not influence QOL or psychological distress after OLT. Within 6 months after OLT, patients with HCV recurrence showed significantly greater Depression (p=0.023), Anxiety (p=0.038), Phobic Anxiety (p=0.001), and Paranoid Ideation (p=0.033) than anti-HCV negative patients. CONCLUSIONS: Liver transplantation improves psychological distress and most, but not all, QOL domains. Recurrent HCV infection is associated with greater psychological distress. PMID- 11059866 TI - Serum thioredoxin levels as an indicator of oxidative stress in patients with hepatitis C virus infection. AB - BACKGROUND/AIM: It has recently been suggested that oxidative stress may be associated with hepatitis C virus (HCV) infection. Thioredoxin (TRX) is a stress inducible thiol-containing protein. The aim of this study was to evaluate the clinical significance of serum TRX levels in patients with HCV-related chronic liver diseases. METHODS: Serum TRX levels were determined with a sandwich enzyme linked immunosorbent assay kit in 174 serum HCV-RNA positive patients, including 6 asymptomatic carriers, 124 chronic hepatitis, 20 liver cirrhosis, and 24 hepatocellular carcinoma, and in 15 healthy volunteers. RESULTS: The serum TRX levels (medians and [ranges], ng/ml) were significantly elevated in the HCV infected patients; 30.9 [20.7-37.7] in asymptomatic carriers, 34.5 [8.6-135.6]* in chronic hepatitis, 42.5 [21.4-97.2]* in liver cirrhosis, and 43.9 [11.7 180.3]** in hepatocellular carcinoma (*p<0.05, **p<0.001, vs. 24.9 [1.3-50.7] in healthy controls). Serum TRX levels were significantly correlated with the serum levels of ferritin and fibrogenesis markers, and with the histological stage of hepatic fibrosis. The serum TRX levels before interferon treatment of patients whose serum HCV-RNA was still positive on day 14 following interferon treatment (42.6 [20.1-90.0]) were significantly higher than those of patients whose serum HCV-RNA was negative on day 14 following interferon treatment (25.8 [7.4-59.8], p<0.05). CONCLUSIONS: The serum TRX levels of patients with HCV infection increased with their serum ferritin levels and the progression of liver fibrosis. Patients with higher serum TRX levels exhibited resistance to interferon therapy. Oxidative stress may therefore be responsible for the pathological mechanism of HCV-related liver diseases and be one of the impediments to eradication of HCV during interferon treatment. PMID- 11059867 TI - Resting energy expenditure in chronic hepatitis C. AB - BACKGROUND/AIMS: Hypermetabolism is considered to be of clinical interest in liver disease and in several chronic viral infections. Whether resting energy expenditure (REE) increases during chronic hepatitis C is not known. Our aims were: (a) to determine the metabolic state of patients with chronic hepatitis C, and (b) to evaluate the effects of interferon therapy on REE. METHODS: Forty seven patients and 20 controls were studied. Sixteen patients failed to respond to interferon and 12 patients stopped the treatment during the first 2 months for various reasons. The 19 responders all received 1 year of interferon. REE (indirect calorimetry) and fat-free mass (FFM, bioelectric impedance analysis) were evaluated before (day 0) and after 90, 180, and 360 days of interferon. The virus load was evaluated in patients before treatment. RESULTS: On day 0, REE expressed as a ratio of FFM (REE/FFM) was higher in patients than in controls (129.2 +/- 14.7 vs 117.9 +/- 9.6 kJ kg FFM(-1) 24 h(-1), p<0.01), and was positively correlated with the viral load (r=0.45, p=0.01). On day 90, REE/FFM had significantly decreased in responders but it did not decrease in non responders (p<0.01). In responders, REE/FFM on days 180 and 360 was similar to that of the controls. CONCLUSIONS: Chronic hepatitis C induces hypermetabolism that is normalized by interferon therapy in responders. The underlying mechanisms of chronic hepatitis C-induced hypermetabolism and its clinical relevance remain to be determined. PMID- 11059868 TI - The relationship of hepatitis B virus infection between adults and their children in Guangxi Province, China. AB - BACKGROUND/AIM: This study aimed to describe the seroepidemiology of hepatitis B virus (HBV) infection, with emphasis on transmission of HBV infection between adults and their children. METHODS: We analyzed the hepatitis sero-survey data collected from 2132 persons aged 1-59 years (624 families) in Guangxi Province, China, 1992. Blood was tested for the presence of the hepatitis B surface antigen (HBsAg), the antibody to hepatitis B core antigen (anti-HBc), and the antibody to hepatitis B surface antigen (anti-HBs). RESULTS: Of the 2132 persons surveyed, 119 (5.6%) reported receiving HBV vaccination. Among those persons who did not receive HBV vaccination, 19% were HBsAg positive (current HBV infection) and 57% had a past HBV infection (they were HBsAg negative and either anti-HBc positive or anti-HBs positive). Among 519 children aged 1-10 years who did not receive HBV vaccination, 21% had current HBV infection and 37% had past HBV infection. Among 289 children of both parents who were HBsAg negative, 16% had current HBV infection and 36% had past HBV infection. CONCLUSIONS: The high prevalence of community-acquired HBV infection in children and the low HBV vaccination coverage in Guangxi should alert public health agencies to re-examine their current policies for preventing HBV transmission. PMID- 11059869 TI - The effects of the conserved extreme 3' end sequence of hepatitis C virus (HCV) RNA on the in vitro stabilization and translation of the HCV RNA genome. AB - BACKGROUND/AIMS: The discovery of an additional 98-base in the extreme 3' end of the hepatitis C virus (HCV) has fueled much speculation as to the role of this sequence on the behavior of the virus. It is now known that this additional 98 base sequence is present and conserved amongst HCV genotypes. This sequence is capable of forming complex and stable high-order structures that may be important in stabilizing the RNA to degradation, facilitating translation and regulating replication of the virus. We have examined the possible role of the HCV extreme 3' end sequence in stabilizing the HCV RNA genome and regulating translation in vitro. METHODS: The extreme 3' end sequence was cloned to downstream of two pre existing two HCV clones: HCV1 (genotype 1a) and HCV-BK (genotype 1b). The reconstructed full-length clones were then tested in vitro for their stability and translation efficiency. RESULTS: We showed that the addition of the conserved 3' end sequence greatly enhanced the stability of HCV1 RNA but had only minimal effect on HCV-BK RNA in mammalian cytoplasmic extracts, suggesting that the requirements for HCV RNA stability vary amongst isolates. Following the optimization of in vitro translation conditions, it was demonstrated that the addition of this 3' end sequence did not affect the translation level from either HCV clone. CONCLUSIONS: The conserved 3' end of the HCV genome confers differential stabilizing effects on two HCV genotype 1 isolates and has no obvious role in the in vitro translation of either clone. PMID- 11059870 TI - Hepatocellular proliferation in patients with chronic hepatitis C and persistently normal or abnormal aminotransferase levels. AB - BACKGROUND/AIMS: Some patients chronically infected with the hepatitis C virus (HCV) have persistently normal alanine aminotransferase (ALT) levels while progressive liver damage is observed histologically. In the present study, we compared the rate of proliferation, apoptosis, and necrosis in liver biopsy specimens of patients with persistently normal or elevated ALT levels. METHODS: Fourteen patients with persistently normal and 14 age- and sex-matched patients with elevated ALT levels were enrolled. Proliferation was detected using anti-Ki 67 in 10-microm liver biopsy specimens of the patients. Apoptosis was measured by TUNEL-assay and by monoclonal anti-M30 directed against caspase-cleaved cytokeratin 18 filaments. RESULTS: The mean number of anti-Ki 67 positive hepatocytes was lower in patients with persistently normal aminotransferases (3.1 +/- 2.8/10(3) vs 10.8 +/- 8.8/10(3) hepatocytes, p<0.0011) and was correlated with serum ALT (r=0.86, p<0.01) and aspartate aminotransferase levels (r=0.83, p<0.01). The rate of apoptosis detected by TUNEL assay was low and not different between patients with persistently normal and elevated aminotransferases. Staining with anti-M30 revealed a granular staining pattern and showed a trend towards higher cell death rates in patients with elevated aminotransferase levels (apoptotic hepatocytes with >75% staining: 3.97 +/- 6.24/10(3) hepatocytes vs 13.65 +/- 19.41/10(3) hepatocytes; p=0.08). CONCLUSIONS: Patients with chronic hepatitis C and normal aminotransferases have significantly lower hepatocyte proliferation rates and show a trend towards lower apoptosis rates compared with patients with elevated aminotransferases. PMID- 11059871 TI - Helicobacter pylori seroprevalence in hepatitis C virus positive patients with cirrhosis. AB - BACKGROUND/AIMS: Liver cirrhosis is a significant cause of death in Italy and one of the most frequent causes of hospitalization. Acute peptic ulcer and upper gastrointestinal bleeding reportedly occur in over one-third of cirrhotic patients. Since Helicobacter pylori (H. pylori) infection strongly correlates with peptic ulcer, we sought to ascertain the prevalence of H. pylori infection in cirrhotic patients. METHODS: In a case-control study, we examined 254 consecutive patients (127 male and 127 female, age range 30-82 years) suffering from hepatitis C virus (HCV)-related cirrhosis and 463 sex- and age-matched patients admitted to the Department of Emergency Care of our hospital (254 male, 209 female, age range 30-79 years) resident in the same area. RESULTS: Antibodies to H. pylori were present in 226/254 (89%) cirrhotic patients and in 275/463 (59%) controls (p<0.0001). The difference was significant both in males and in females. CONCLUSIONS: The very high prevalence of H. pylori infection may explain the frequent occurrence of gastroduodenal ulcer in cirrhotic patients and may possibly determine the prognosis of those who are also infected with HCV. PMID- 11059872 TI - Cost-effectiveness of combination therapy for naive patients with chronic hepatitis C. AB - BACKGROUND/AIMS: The higher initial cost of combination therapy is a factor which may discourage its use in naive patients with histologically mild or moderate chronic hepatitis C. However, chronic hepatitis C is a slowly progressive disease associated with a decrease in life expectancy and quality of life. The objective of this study was to determine if the incremental sustained response rate of combination therapy is sufficient to outweigh its extra cost. METHODS: Chronic hepatitis C progression was studied using a Markov model in which cohorts of patients were treated with combination therapy for 6-12 months or with interferon for 12 months. The sustained virological response rates applied were 43, 35 and 19%, respectively, for combination therapy for 12 months, for 6 months and for interferon for 12 months. Costs for each clinical state were calculated according to clinical practice in Spain. RESULTS: In a 30-year-old patient with moderate chronic hepatitis C, combination therapy for 12 months increases life expectancy by 4.1 years compared with interferon for 12 months. In mild disease, the increase in life expectancy is lower. The cost per life-year saved in patients with chronic hepatitis C ranges from 880 to 64.421 euros depending on the age of the patient, the degree of hepatic lesion and the type and duration of therapy. Compared to other treatments accepted as standard practice in other therapeutic areas, combination therapy for chronic hepatitis C is cost-effective. CONCLUSIONS: In patients of any age with moderate chronic hepatitis C and those with mild disease under 50 years of age, combination therapy for 12 months is the most cost-effective schedule, whereas in older patients with mild hepatitis, combination therapy for 6 months is the preferred option based on cost effectiveness criteria. PMID- 11059873 TI - The role of magnetic resonance cholangiography in primary sclerosing cholangitis. PMID- 11059874 TI - Diagnosis of primary sclerosing cholangitis--autoimmune hepatitis overlap syndrome: to score or not to score? PMID- 11059875 TI - Combination therapy in naive patients with chronic hepatitis C: more value for the money. PMID- 11059876 TI - Radiofrequency ablation in the treatment of hepatocellular carcinoma--a clinical viewpoint. PMID- 11059877 TI - Liver nodules resembling focal nodular hyperplasia after hepatic venous thrombosis. AB - We report the case of focal nodular hyperplasia-like nodular hepatic lesions, that developed in the liver of a 35-year-old Caucasian female who required orthotopic liver transplantation for Budd-Chiari syndrome. The rapid development of focal nodular hyperplasia-like lesions in a severe hepatic vascular disorder and in the absence of cirrhosis may represent an additional argument in favor of the vascular origin of focal nodular hyperplasia. The pathogenesis of the nodules is not clear, but pathological arterialization of the liver in hepatic vein thrombosis may be a candidate mechanism. PMID- 11059878 TI - Successful treatment of hepatitis B virus associated polyarteritis nodosa with a combination of prednisolone, alpha-interferon and lamivudine. AB - Therapy of hepatitis B virus (HBV)-associated poly-arteritis nodosa is still evolving. Here we report a successful treatment with a short-term steroid administration in combination with a-interferon and lamivudine and a complete sequence analysis of the HBV genome. A 58-year-old man presented with the symptoms of mononeuritis multiplex associated in time with the onset of highly replicative hepatitis B. Polyarteritis nodosa was confirmed by biopsy. During an initial course with alpha-interferon and prednisolone no clinical improvement or hepatitis B virus seroconversion was observed. After addition of lamivudine to the protocol with fast tapering of prednisolone, HBV DNA fell to undetectable levels within 1 month and liver transaminases normalized. After 6 months of treatment HBeAg seroconversion took place, followed by HBsAg seroconversion 2 months later. Clinical symptoms of polyarteritis improved. No relapse of polyarteritis or hepatitis B was seen during the follow up of 9 months. Complete sequence analysis of the HBV genome revealed 6 nucleotide mutations but none in a relevant antigenic epitope. The present protocol of short-term prednisolone administration combined with alpha-interferon and lamivudine was effective for the treatment of HBV-related polyarteritis nodosa and may be a promising new therapeutic approach. PMID- 11059879 TI - Hepatic angiomyolipoma: a rarely described tumor. PMID- 11059880 TI - Is it useful to perform the RNA test for hepatitis C in health care workers after an accidental needlestick? PMID- 11059881 TI - Development and application of bioartificial skin. PMID- 11059882 TI - Dialysate flow distribution in hollow fiber hemodialyzers with different dialysate pathway configurations. AB - The efficiency of a hemodialyzer is largely dependent on its ability to facilitate diffusion, since this is the main mechanism by which small solutes are removed. The diffusion process can be impaired if there is a mismatch between blood and dialysate flow distribution in the dialyzer. The objective of the paper was to study the impact of different dialysate compartment designs on dialysate flow distribution and urea clearances. Eighteen hollow fiber 1.3 m2 hemodialyzers were studied, 6 each of 3 designs: Type A--standard fiber bundle (PAN 65DX Asahi Medical, Tokyo, Japan); Type B--spacing filaments external to the fibers (PAN 65SF Asahi Medical, Tokyo, Japan); Type C--fibers waved to give Moire structure (FB130 Nissho-Nipro, Osaka, Japan). IN VITRO STUDIES: 3 dialyzers of each type were studied following dye injection into the dialysate compartment. Dynamic sequential imaging of longitudinal sections of the dialyzer were undertaken, using a new generation helical CT scanner (X-Press/HS1 Toshiba Corporation, Tokyo, Japan). In vivo studies: 3 dialyzers of each type were studied, in randomized sequence, in 3 different patients under standardized dialysis conditions. Blood- and dialysate-side urea clearances were measured at 30 and 150 minutes of treatment. Macroscopic and densitometrical analysis revealed that flow distribution was most homogeneous in the dialyzer with Moire structure (Type C) and least homogeneous in the standard dialyzer (Type A). Space yarns (Type B) gave an intermediate dialysate flow distribution. Significantly increased urea clearances (p<0.001) were seen with Types B and C, compared to the standard dialyzer. Type C (Moire) had the highest clearances although these were not significantly greater than Type B (space yarns). In conclusion, more homogeneous dialysate flow distribution and improved small solute clearances can be achieved by use of spacing yarns or waved (Moire structure) patterns of fiber packing in the dialyzer. These effects are achieved probably as a result of reduced dialysate channeling resulting in a lower degree of mismatch between blood and dialysate flows. The new radiological technique using the helical CT scanner allows detailed flow distribution analysis and has the potential for testing future modifications to dialyzer design. PMID- 11059883 TI - Blood flow around hollow fibers. AB - In an artificial lung, blood is oxygenated and flows around a bundle of hollow fibers while gas flows inside the fiber. The objective of this study is to understand the hydrodynamics of three different fiber banks (inline square IS, staggered square SS and equilateral triangle ET) and to investigate the influence of both a Newtonian and non-Newtonian Casson viscosity model on the flow field. A two-dimensional finite element model for permanent, isothermal, laminar blood flow perpendicular to hollow fibers is used. All fibers are assumed identical, straight and parallel. Porosity ranges from 0.4 to 0.6 and Reynolds number varies from 1 to 60. For a given Re, ET generates less resistance than SS, the latter being comparable with IS. A lower porosity increases resistance. Non-Newtonian viscosity affects velocity patterns only at low Re (<0.5) and higher porosity (>0.5). Resistance at low Re is significantly elevated in the fiber banks due to an overall increase in viscosity. This model makes it possible to study the influence of geometry and viscosity on hydrodynamics in fiber banks and may aid in the optimization of hollow fiber artificial lung design. PMID- 11059884 TI - Is there a role for hemoperfusion/hemodialysis as a treatment option in severe tricyclic antidepressant intoxication? AB - OBJECTIVE: Suicidal self-poisoning with tricyclic antidepressants like doxepin is a major therapeutic problem in emergency medicine with a high fatality rate. Deaths are mainly caused by cardiotoxicity with arrhythmias, intraventricular conduction disturbances and myocardial depression. For treatment, alkalinization and hypertonic saline are recommended. The role of extracorporeal, treatment procedures is not clear. The possible benefit of hemoperfusion/hemodialysis is discussed in a case report with respect to the published literature. CASE REPORT: After ingestion of an amount of at least 5000 mg doxepin a 37-year-old man with endogenous depression developed cardiac arrest. After preclinical resuscitation with prolonged external cardiac massage, he was admitted to the intensive care unit with persistently severe hypotension and wide QRS complexes (230-260 ms). Despite fluid load, alkalinization, hypertonic saline and high-dose vasoactive substances the patient's condition did not improve. Hemoperfusion over hemoresin combined with hemodialysis led to an impressive clinical improvement with shortening of QRS duration (from 230 to 120 ms) and hemodynamic stabilization. The patient fully recovered without neurologic deficits. CONCLUSION: We report a successful treatment with hemoperfusion over hemoresin and hemodialysis in a patient with life-threatening doxepin poisoning intractable with the generally recommended treatment. In such acute TCA intoxication with severe cardiotoxicity, hemoperfusion/hemodialysis should be considered a potential treatment option, as the "toxicokinetics" of drugs may totally differ from their usual pharmacokinetic behaviour. Experimental and clinical studies are needed to clarify the toxicokinetics of TCA in order to improve the therapeutic approach. PMID- 11059885 TI - Tissue-engineered heart valve leaflets: an effective method for seeding autologous cells on scaffolds. AB - BACKGROUND: A precondition for the successful formation of tissue-engineered heart valves is the generation of a proper matrix on biodegradable scaffolds over a limited period of time. The aim of this study was to find an effective method of seeding autologous cells on these scaffolds to create a new matrix for heart valves. METHODS: Myofibroblasts and endothelial cells were isolated and cultured from an ovine artery. A synthetic biodegradable scaffold consisting of polyglycolic and polylactic acids was seeded first with the myofibroblasts, then coated with endothelial cells. Three different methods of myofibroblast seeding were compared: I) daily seeding of myofibroblasts (1x10(6)) for ten days and culture for four days; II) seeding of myofibroblasts (1x10(7)) and culture for 14 days with the use of a simple medium; III) seeding of myofibroblasts (1x10(7)) with the use of a medium containing collagen and culture for 14 days. Light and electron microscopic analyses were performed. RESULTS: The group that used the medium containing collagen showed the best results in terms of seeding efficiency. CONCLUSION: Seeding autologous cells with a medium containing collagen onto the scaffold showed the largest cell population and might generate the best matrix on the scaffold. PMID- 11059886 TI - Frequent replacement and conventional daily wear soft contact lens symptomatic patients: tear film and ocular surface changes. AB - We analyzed tear film changes, ocular surface tissue alterations and inflammatory conditions in Frequent Replacement Soft (FRS) Contact Lens (CL) or Conventional Soft (CS)-CL wearers suffering from eye discomfort, by applying complementary diagnostic tests. Our data suggest the following conclusions: i) eye discomfort symptoms begin after 1-3 years of successful wear, mainly in FRS-CL wearers; ii) tear film stability is affected by CL wear, mainly in FRS-CL wearers; iii) CL wear produces a subclinical moderate inflammation based on mononuclear cell recruitment in 3 out of 4 patients of both types of lenses; iv) CL wear induces dry eye conditions, more pronounced in FRS-CL users; v) discomfort symptoms, presence of subclinical inflammation, degree of dryness are not directly related to a clinical presentation of intolerance, mainly in FRS-CL wearers; vi) CL wear related discomfort symptoms should never be underestimated, since a hidden inflammation or dryness can be present beyond an apparently good clinical picture. PMID- 11059887 TI - DALI LDL-apheresis: anticoagulation with r-hirudin in a patient with heparin induced thrombocytopenia (HIT II). AB - A 50-year old male patient with familial hypercholesterolemia and hyperlipoproteinemia (a), who underwent low density lipoprotein-apheresis treatment developed heparin-induced thrombocytopenia type II (HIT II). Because heparin is contraindicated in patients with HIT, an alternative LDL-apheresis system and modified anticoagulation regimen was necessary. Treatment was changed to a new system called DALI (direct adsorption of lipids). After confirmation of the diagnosis HIT II, DALI LDL-apheresis was carried out with recombinant-hirudin (lepirudin) and citrate in order to prevent hypercoagulability. Efficient LDL apheresis therapy with minimum therapeutic blood levels of lepirudin (1.4 mg/dl) was achieved with an initial intravenous bolus of 0.114 mg/kg of lepirudin followed by continuous lepirudin infusion of 0.350 mg/h. Thrombin-antithrombin III complex production was well controlled and other hemostatic markers showed no abnormalities. LDL-cholesterol and lipoprotein(a) concentrations were effectively reduced. R-hirudin offers a novel anticoagulation strategy and is, at present, the only alternative for patients with HIT II requiring LDL-apheresis on a regular basis. PMID- 11059888 TI - Acute intravascular hemolysis in two patients transfused with dry-platelet units obtained from the same ABO incompatible donor. AB - Since 1989 we have been collecting dry-platelets on a routine basis. Dry platelets are those collected along with 25-30 ml of contaminating plasma cell with separators such as the Amicus, AS 104 and the Excel Pro. Platelets are resuspended in non plasma media for storage and for at least 60 hours their viability and functionality are not impaired. In this article we report on two hemolytic crises determined by two O Rh D + units of single donor platelets (SPD) taken from the same donor in a double-apheresis session. The two split units were administered to two A Rh D + patients suffering from metastatic breast cancer and severe aplastic anemia (SAA) respectively. In both cases the hemolytic reaction was of the intravascular type, with a drop in hemoglobin (Hgb) level from 8.6 to 5.4 and from 8.8 down to 5.3 g/dl respectively. From the patients' RBC only alpha agglutinins were eluted and donor's indirect antiglobulin test (IAT) was negative with extended panel RBCs. In the first case the clinical course after erythroexchange (Erex) was uneventful whereas in the second one, that suffering from SAA, after Erex, acute renal failure and shock did complicate the clinical course and the patient died seven days after the incriminated platelet transfusion. PMID- 11059889 TI - A physical model of the human systemic arterial tree. AB - A physical model of the human arterial tree has been developed to be used in a computer controlled mock circulatory system (MCS). Its aim is to represent systemic arterial tree properties and extend the capacity of the MCS to intraortic balloon pump (IABP) testing. The main problem was to model the aorta simply and to accurately reproduce aortic impedance and related flow and pressure waveforms at different sections. The model is composed of eight segments; lumped parameter models are used for its peripheral loads. After the numerical simulation, the physical model was reproduced as a silicon rubber tapered tube. This rubber was chosen for its stability over time and the acceptable behaviour of its Young's modulus (Ey = 22.23 gf x mm(-2)) with different loads and in comparison with data from the literature (Ey approximately 20.4 gf x mm(-2)). The properties of each segment of the aorta were defined in terms of compliance, resistance and inertance as a function of length, radius and thickness. The variable thickness was obtained using positive and negative molds. Total static compliance of the aorta model is about 1.125 x 10(-3) g(-1) x cm4 x sec2 (1.5 cm3 x mmHg(-1)). Measurements were performed both on numerical and physical models (in open and closed loop configuration). Data reported show pressure and flow waveforms along with input impedance modulus and phase. The results are in good agreement with data from the literature. PMID- 11059890 TI - XXVII ESAO Congress. European Society for Artificial Organs. CHUV Lausanne, Switzerland, August 31-September 2, 2000. Summary of the meeting. PMID- 11059891 TI - Hiroshi Asanuma (1926-2000). PMID- 11059892 TI - Calcium binding proteins in motoneurons at low and high risk for degeneration in ALS. AB - Recent reports challenge the hypothesis that expression of calcium binding proteins contributes to the greater resistance of some motoneurons to degeneration in amyotrophic lateral sclerosis (ALS). We therefore re-examined, using immunohistochemistry, the expression of calbindin, calretinin and parvalbumin in vulnerable (hypoglossal, XII; and cervical spinal) and resistant (oculomotor, III) motoneurons of adult rats. Calbindin immunoreactivity was lacking in motor nuclei but strong in the dorsal horn. Calretinin was expressed in spinal, but not III or XII, motoneurons. Parvalbumin immunoreactivity, tested with a polyclonal antibody, was intense in spinal and III, but not XII, motoneurons; however, no staining in the ventral horn was observed with a monoclonal antibody. Differential expression of calretinin and parvalbumin within vulnerable motoneurons suggests that immunoreactivity for these proteins is not a reliable marker for resistance to degeneration in ALS. PMID- 11059893 TI - Time course of focal slow wave activity in transient ischemic attacks and transient global amnesia as measured by magnetoencephalography. AB - In this longitudinal study multichannel MEG was used to localize and to quantify focal pathological spontaneous neuromagnetic activity in six patients with transient ischemic attacks (TIA) and two patients with transient global amnesia (TGA). Slow (2-6 Hz) and beta (14-30 Hz) activity were monitored up to 10 weeks. Results were compared with normative data, and changes over time were statistically analyzed. MEG detected pathological activity that persisted clinical symptoms. Focal slow activity originating from sensorimotor (TIA) and mesiotemporal (TGA) cortices exceeded normal values up to 14 times during the first hours after the attack and recovered to normal within 11 days. Focal beta activity was not useful to monitor the time course of TIA or TGA. PMID- 11059894 TI - Sonic hedgehog promotes proliferation and tyrosine hydroxylase induction of postnatal sympathetic cells in vitro. AB - The role of Sonic hedgehog (shh) in neural crest development was initially suggested by its involvement in patterning of the neural tube. While largely implicated in cell fate determination during development, its recently discovered role in the development of neurons postnatally prompted the possibility that neural crest derivatives of the sympathoadrenal lineage may respond to Shh postnatally. In the present study, we show that Shh promotes proliferation of postnatal sympathetic cells in culture. While it has been previously found to induce tyrosine hydroxylase (TH) production in the developing midbrain, we also demonstrated that Shh is capable of promoting TH induction of mature sympathetic neurons in vitro. This duality in Shh can be inhibited by activation of protein kinase A. These findings suggest that cell response to Shh is conserved in sympathetic ganglia derived from the neural crest, and further supports the notion that Shh can function postnatally in a dose-dependent manner to mediate neuronal cell fate. PMID- 11059895 TI - Exposure to pulsed high-frequency electromagnetic field during waking affects human sleep EEG. AB - The aim of the study was to investigate whether the electromagnetic field (EMF) emitted by digital radiotelephone handsets affects brain physiology. Healthy, young male subjects were exposed for 30 min to EMF (900 MHz; spatial peak specific absorption rate 1 W/kg) during the waking period preceding sleep. Compared with the control condition with sham exposure, spectral power of the EEG in non-rapid eye movement sleep was increased. The maximum rise occurred in the 9.75-11.25 Hz and 12.5-13.25 Hz band during the initial part of sleep. These changes correspond to those obtained in a previous study where EMF was intermittently applied during sleep. Unilateral exposure induced no hemispheric asymmetry of EEG power. The present results demonstrate that exposure during waking modifies the EEG during subsequent sleep. Thus the changes of brain function induced by pulsed high-frequency EMF outlast the exposure period. PMID- 11059896 TI - Learning effects on event-related brain potentials. AB - This study investigates whether learning can manifest itself in EEG changes without explicit awareness of the acquired knowledge and without any motor requirement (to rule out acquisition of motor skills). Subjects passively heard particular stimulus combinations interspersed with random combinations. Immediately after this, stimulus combinations were presented which contained violations of the previously learned regular sequences. These violations elicited two frontally distributed negative waves peaked at about 250 ms and 500 ms, respectively. None of the participants could detect any regularity of the stimulation. No significant difference between regular vs random stimulus combinations was found during acquisition. These findings indicate that learning without explicit knowledge does not require motor activity (i.e. is independent of learning response sequences) and does not depend on stimulus probabilities. PMID- 11059897 TI - Changes in the apparent diffusion coefficient of water and T2 relaxation time in gerbil hippocampus after mild ischemia. AB - Selective neuronal death of the hippocampal CA1 area after mild ischemia is known as delayed neuronal death (DND). Progression of DND was evaluated using 7T MRI in gerbils. In the CA1, the T2 relaxation time started to increase on day 3 and was significantly higher on day 4 than that of the control gerbils. However, the apparent diffusion coefficient of water (ADC) was significantly lower on day 1 than in the control and continued to decline up to day 4. Changes in T2 coincided with loss of MAP2 immunoreactivity, while the ADC decrease preceded these changes. After 1 month, the ADC had returned to within the normal range, while T2 had decreased to below the control level. These results suggest that MRI is useful for monitoring DND. PMID- 11059898 TI - Coincidence of ipsilateral ocular dominance peaks with orientation pinwheel centers in cat visual cortex. AB - Geometrical relationships among multiple cortical maps, such as those between ocular dominance and orientation maps, are a prominent feature of the brain's functional architecture. It is also well known that there is a strong bias of cortical responses toward the contralateral eye during early postnatal development. We wondered therefore whether and how such an imbalance of cortical responsiveness in a developing animal might influence the mutual geometrical relationships between orientation and ocular dominance maps in adult animals. The results of our study indicate the existence of a strong tendency for the peaks of the ipsilateral eye domains to coincide with the location of point singularities (pinwheel centers) in orientation maps. No such relationship was found for the peaks of contralateral eye domains. Computational studies reproduced similar asymmetry in the coincidence under the contralateral eye bias of inputs. Our study raised the idea that the pinwheel centers play an important role for retaining the weaker ipsilateral eye inputs during normal development. PMID- 11059899 TI - Peripheral axotomy increases cholecystokinin release in the rat anterior cingulate cortex. AB - The anterior cingulate cortex (ACC) is a limbic region with a high density of cholecystokinin (CCK) immunoreactivity, that has been suggested to be of importance for the affective and emotional component of pain. In the present microdialysis study, performed in the awake rat, we demonstrate a bilateral 4- to 6-fold increase of the potassium-induced release of CCK-like immunoreactivity (CCK-LI) in the ACC 2-3 weeks after a unilateral transection of the sciatic nerve (axotomy), an animal model of phantom limb or deafferentiation pain. Considering the implication of CCK in pain modulation and anxiety, we suggest that an altered activity of CCK containing neurons in the ACC may be of importance for the affective component of certain pain conditions. PMID- 11059900 TI - Activation of kappa opioid receptors in the rostral ventromedial medulla blocks stress-induced antinociception. AB - Prior work has shown that kappa opioids may attenuate the effects of analgesic mu receptor agonists in some neural circuits related to pain modulation. This study examined whether hypoalgesia following exposure to a signal for shock is attenuated by infusions of the kappa agonist U69593 into the rostral ventromedial medulla (RVM). Rats were trained with paired or unpaired presentations of white noise and foot shock. On test days, tail flick latencies were measured before, during, and after exposure to the auditory conditioned stimulus (CS). One of three doses of U69593 (0.0445, 0.178 and 1.00 microg) or an equivalent volume of saline was injected into the RVM. Rats that had received noise-shock pairings displayed conditional hypoalgesia (CHA) compared to those given unpaired presentations. Expression of CHA was completely blocked by the highest dose of U69593 (1.00 microg) injected 20 min before testing, indicating an antagonistic effect of U69593 on expression of CHA. These findings are discussed in terms of the evidence for antagonism of morphine- and DAMGO-induced hypoalgesia by kappa agonists. PMID- 11059901 TI - Neurobehavioural effects of hypergravity conditions in the adult mouse. AB - To evaluate the behavioural response to a hypergravity condition in CD-1 mice, young adult subjects of both sexes were exposed to 2 g for a single 60 min rotational session. Motion sickness (MS) and ethological-type scoring of different activities were used to evaluate the behavioural response. Nerve growth factor (NGF) and brain derived neurotrophic factor (BDNF) levels were also assessed. Behavioural scores indicated a transient mild sickness associated with hypergravity, with reduction in spontaneous activity. In males kaolin consumption (a MS index) increased following rotation while females consumed more kaolin irrespective of whether they have been rotated or simply exposed to the noise and vibration of the rotational apparatus. In males, hypothalamic NGF levels were markedly increased after rotation while no major changes were observed in central BDNF expression. These results indicate mice may represent a suitable MS model. PMID- 11059902 TI - Toxic effect of the beta-amyloid precursor protein C-terminus fragment and Na+/Ca2+ gradients. AB - There is evidence to suggest that certain metabolic fragments of the beta-amyloid precursor protein (betaAPP) containing the whole of the beta-amyloid (Abeta) sequence are toxic to cells. We showed previously that the 105-amino acid C terminal peptide (CT105) fragment, incorporating Abeta, is particularly toxic to Xenopus oocytes as well as to mammalian neurons. Here, we investigated the contributions of Na+ and Ca2+ gradients to intracellular CT105-induced toxicity in oocytes, monitored by measuring the membrane resting potential. The concentration gradients of Na+ and Ca2+ were manipulated to determine the involvement of the trans-membrane concentration gradients of these ions in the mode of action of CT105. The results suggested that Na+ influx and intracellular events are mainly responsible for the observed CT105-induced toxicity. PMID- 11059903 TI - Expression of 5-HT1A receptor mRNA in rat dorsal raphe nucleus and ventrolateral periaqueductal gray neurons after peripheral inflammation. AB - In the present study we observed the expression of 5-hydroxytryptamine (5-HT)1A receptor mRNA in the dorsal raphe nucleus (DRN) and ventrolateral periaqueductal gray (vlPAG) neurons, especially in 5-HT immunoreactive neurons (5-HT-IR), using in situ hybridization (ISH) and double staining with fluorescent ISH (FISH) and immunohistochemical (FIH) techniques. The findings of this study demonstrated that 5-HT1A receptor mRNA was expressed with moderate to high level in the DRN and vlPAG neurons. Following carrageenan inflammation, the expression of 5-HT1A receptor mRNA in the DRN and bilateral vlPAG neurons was significantly increased. The peak occurred at 3-8h followed by a clear decrease at 24 h, which basically corresponded to the time-course of behavioral hyperalgesia. Moderate 5-HT1A receptor mRNA and 5-HT immunoreactive (5-HT-IR) double-labeled cells were observed in the DRN and vlPAG, suggesting that some of 5-HT1A receptors in the DRN and vlPAG may be autoreceptors. Eight hours after carrageenan injection, the number of the double labeled cells was significantly increased. These results suggest that the synthesis of 5-HT1A receptors, including autoreceptors, is increased in the DRN and vlPAG during peripheral inflammation. PMID- 11059904 TI - Estimation of striatal dopamine spillover and metabolism in vivo. AB - A microdialysis probe with an attached microinjection cannula was inserted into rat striatum. [3H]dopamine (or [14C]sucrose as a reference substance for diffusion) was infused via the cannula, with microdialysate sampled for concentrations of endogenous and [3H]-labeled dopamine and its metabolites. The calculated specific activities of the [3H]-labeled metabolites led to the conclusions that striatal extracellular dopamine undergoes inactivation mainly by extraneuronal but also by neuronal uptake and intracellular metabolism. Some of the dopamine taken up into nerve terminals slowly re-enters (spillover) the extracellular fluid unchanged. This spillover was calculated to be about 5 pmol/min. Destruction of dopaminergic terminals increases the turnover of vesicular stores in the surviving terminals, both by increased vesicular leakage and by increased release into the extracellular fluid. PMID- 11059905 TI - Transection of dysmyelinated optic nerve axons in adult rats lacking myelin basic protein. AB - Injury to myelin or oligodendrocytes may manifest as dysmyelinating or demyelinating conditions of the CNS. Previous studies using dysmyelinated animal models (myelin basic protein mutants) suggest possible axonal dysfunction with complete loss of myelin. In this present study, we evaluated retinal ganglion cell survival after axotomy in MBP mutants to determine if prolonged dysmyelination of CNS axons exerted a detrimental effect on neuronal survival. We demonstrated that the survival of retinal ganglion cells with dysmyelinated axons is identical to retinal ganglion cells with myelinated axons after survival times up to 180 days. In myelin diseases where axon transection is a consistent consequence of demyelination resulting in progressive neurological deterioration, the absence of myelin does not accelerate neuronal death. PMID- 11059906 TI - Glucocorticoids modulate BDNF mRNA expression in the rat hippocampus after traumatic brain injury. AB - Brain-derived neurotrophic factor (BDNF) expression in rat hippocampus is increased after experimental traumatic brain injury (TBI) and may be neuroprotective. Glucocorticoids are important regulators of brain neurotrophin levels and are often prescribed following TBI. The effect of adrenalectomy (ADX) on the expression of BDNF mRNA in the hippocampus after TBI has not been investigated to date. We used fluid percussion injury (FPI) and in situ hybridization to evaluate the expression of BDNF mRNA in the hippocampus 4 h after TBI in adrenal-intact or adrenalectomized rats (with or without corticosterone replacement). FPI and ADX independently increased expression of BDNF mRNA. In animals undergoing FPI, prior ADX caused further elevation of BDNF mRNA and this upregulation was prevented by corticosterone replacement in ADX rats. These findings suggest that glucocorticoids are involved in the modulation of the BDNF mRNA response to TBI. PMID- 11059907 TI - Cytokine production in unmedicated and treated schizophrenic patients. AB - Recent findings have strengthened the hypothesis that immunological dysfunctions may contribute towards the multifactorial pathogenesis of schizophrenia. The validity of these findings is questioned by the fact that most studied subjects have received potentially immunomodulatory medication upon investigation. In order to rule out such confounding effects, 24 initially unmedicated acutely ill schizophrenic patients were studied immunologically and psychiatrically (PANSS) before and during 4 weeks of neuroleptic treatment. The production of IFN-gamma was decreased upon admission and after 2 weeks of treatment compared to matched healthy controls. No differences in IL-2 and IFN-gamma production between unmedicated and medicated states were observed. These results do not support the notion that neuroleptic medication in vivo might influence TH1 cytokine production in schizophrenia. PMID- 11059908 TI - CGMP increases in satellite cells of nitric oxide synthase-containing sensory ganglia. AB - cGMP and the enzymes, nitric oxide synthase (NOS) and heme oxygenase-1 (HO-1), the products of which stimulate soluble guanylyl cyclase activity, were investigated in cultured dorsal root ganglia (DRG), and nodose ganglia of adult rats. A dramatic increase of cGMP-positive satellite cells in ganglia cultured for 24 or 48 h was observed, particularly in Th8-L2 DRG and in nodose ganglia. These ganglia also contained most NOS-positive neurones, as reflected by NADPH diaphorase histochemistry. HO-1 immunoreactivity increased in satellite cells, but in different cells to those in which cGMP increased. These results suggest that both NO and CO could be involved in signalling between neurones and satellite cells in sensory ganglia during regeneration. PMID- 11059909 TI - Role of cholinergic and GABAergic systems in the feedback inhibition of dorsal raphe 5-HT neurons. AB - Several observations indicate that 5-HT1A receptors found on a long neuronal feedback loop, originating from the medial prefrontal cortex, regulate 5-HT neuronal firing. In the present study, the muscarinic (M) receptor antagonists atropine and scopolamine as well as the M2 receptor antagonist AF-DX 116, but not the preferential M1 receptor antagonist pirenzepine, reduced the suppressant effect of the 5-HT1A receptor agonist 8-OH-DPAT on the spontaneous firing activity of rat dorsal raphe 5-HT neurons. Moreover, AF-64A-induced lesions of cholinergic neurons directly in the medial prefrontal cortex and after its i.c.v. injection attenuated the effect of 8-OH-DPAT. Finally, the NMDA receptor antagonist (+)MK-801 and the GABA(B) receptor antagonist SCH-50911, but not the GABA(A) receptor antagonist (-)bicuculline, dampened the latter response. The present study unveiled a key role for the cholinergic and GABAergic systems in the feedback inhibition of dorsal raphe 5-HT neurons. PMID- 11059910 TI - Enhanced GABA(A) inhibition enhances synchrony coding in human perception. AB - The benzodiazepine, lorazepam enhances the efficiency of local, inhibitory GABA(A) synapses in the cortex, which stabilize postsynaptic, excitatory activity by synchronizing their own discharges at around 40 Hz. Treatment with lorazepam has also been shown to adversely influence detection performance in perceptual tasks, suggesting a role for GABA(A)-mediated synchronization during visuo perceptual organization. Consistent with these findings we report that reaction times to target stimuli were slower following lorazepam treatment. However, when targets followed presentation of a synchronized prime, presented within a flickering 40-Hz display matrix, the effects of priming were amplified relative to baseline and control conditions. We conclude that enhanced GABA(A)-induced inhibition enhances stimulus-evoked synchronization with differential effects upon mechanisms of perceptual segmentation and grouping. PMID- 11059911 TI - Perinatal organization of a sexually dimorphic aromatase enzyme-containing immunoreactive nucleus. AB - Sexual dimorphisms have been identified in the brains of many vertebrates. The adult musk shrew brain contains a sexually dimorphic nucleus of aromatase (AROM) immunoreactive (ir) neurons within the medial preoptic area (mPOA). This cell group is larger and contains more neurons in males than in females. We examined the effects of perinatal hormone manipulations on the organization of this nucleus. Administration of testosterone (T) to female pups (days 1-5 postnatal) increased the size of the AROM-ir nucleus. Males castrated on postnatal day 1 had a smaller AROM-ir nucleus as adults than did males that remained gonadally intact during development. These data show that T and/or its estrogenic metabolite act during an early critical period to organize the development of AROM-ir neurons in the mPOA. PMID- 11059912 TI - Firing patterns of pallidal cells in parkinsonian patients correlate with their pre-pallidotomy clinical scores. AB - It is unclear how the disordered activity of cells in the basal ganglia contributes to the symptoms of Parkinson's disease (PD). We recorded from single neurons extracellularly in 3 regions of the globus pallidus (GPe, GPie and GPii) in patients undergoing pallidotomy for PD. Movement-related cell firing patterns, analysed using hidden Markov models, were significantly correlated with patients' preoperative clinical scores (off drugs). Responses of cells in GPii correlated best with the scores for specific motor tasks, rather than general ones related to activities of daily living, but the reverse was true for responses from GPe. In both GPii and GPe, a higher score (i.e. greater parkinsonian severity) was associated with greater variability in cell firing rather than an increase in firing rate itself. PMID- 11059913 TI - ECG of the Month. Give P's a Chance. Incomplete AV dissociation. PMID- 11059914 TI - Medical management of pediatric chronic sinusitis. AB - Pediatric sinusitis can be a challenging disease to treat, whether by a primary care physician or an otolaryngologist. When initial appropriate therapy fails to resolve the disorder, frustration may develop on the part of the patient, the family, and the physician. In addition to treatment with appropriate antibiotics for a sufficient length of time, other associated conditions that can exacerbate the condition must be considered and addressed as necessary. These may include viral upper respiratory infections, allergic rhinitis, immune deficiencies, asthma, and gastroesophageal reflux disease. Unless all associated conditions have been optimized, treatment of chronic sinusitis will often be unsuccessful. Recognition that there may be another factor contributing to the patient's continuing illness should prompt appropriate evaluation and occasionally referral to appropriate specialists. Except for the unusual pediatric patient with a truly anatomic disorder or an underlying chronic illness such as cystic fibrosis, proper medical management will almost always resolve chronic sinusitis. PMID- 11059915 TI - Radiology case of the month. A groin mass. Adductor muscle pseudotumor. PMID- 11059916 TI - Walker Percy's Magic mountain. PMID- 11059917 TI - Adolescent violence. PMID- 11059918 TI - Proven practices for reducing aggressive and noncompliant behaviors exhibited by young children at home and at school. AB - One of the single most powerful predictors of aggressive and noncompliant behaviors exhibited in early childhood is coercive parent-child interaction. Coercive parent-child interaction has been linked to multiple negative outcomes in the lives of children. When children learn to relate to their parents and the world in the context of coercive interaction, they are likely to experience significant deficits in the prosocial skills critical to school success. These children are much more likely to experience school failure and teacher and peer rejection. Further, when noncompliant and aggressive children enter school, they are most frequently exposed to a series of ineffective and increasingly restrictive treatments. Proven strategies exist to teach parents and children prosocial ways of interacting and to address these problems in the classroom, but in many cases these types of services are not easily accessible or routinely available. This paper makes recommendations for identifying effective, proven treatment strategies when practitioners observe coercive parent-child interaction or child noncompliance and aggression. PMID- 11059919 TI - A developmental psychopathology approach to understanding and preventing youth violence. AB - There are many views in both the lay and professional literatures as to the causes of violent behavior. These views influence the types of interventions that are designed and tested for preventing violence or for treating violent individuals. In this paper, the author provides a developmental psychopathology framework in which violent behavior is viewed as a developmental outcome that can result from many different pathways, each involving a somewhat different interaction of causal processes. This way of viewing violent behavior has already helped to guide some of the more effective prevention and treatment strategies, with the key to their success being a comprehensive and individualized approach to intervention. This approach for understanding violent behavior also points the way to some important goals for a next generation of prevention and treatment programs. PMID- 11059920 TI - The effects of community violence exposure on Louisiana's children. PMID- 11059921 TI - Violence prevention: myth or reality? AB - Newspapers and television daily attest to the fact that violence is a pervasive element in our society, especially among our youth. We have come to a point in this nation where we see violence everywhere. It is on the streets, in the workplace, and especially in the schools. How did this happen to our society? Is this unrivaled period of juvenile violent crime a new phenomenon? Our society demands that children have a safe environment in which to learn and grow, yet continued reports of youth violence indicates that our efforts have not been successful. Is violence prevention a myth or reality? Since children are our future, how can we provide them with the skills that will afford them the opportunity to become productive members of society? PMID- 11059922 TI - Children, adolescents, and guns in Louisiana: a thought experiment. AB - More children and youth in Louisiana die from firearm injuries than from any other injury, including motor vehicle accidents. Many survive their injuries to lead lives with permanent disabilities. The cost to the victims and to society as a whole is enormous. What is the best way to address this issue? What is the physician's role, both as an individual and as a member of a medical organization? This paper describes a way of thinking about firearm injury prevention. It introduces the reader to the Haddon Matrix and the Intervention Decision Matrix. It then reviews six options and offers one model, motor vehicle injury reduction, as a way to consider intervention options. PMID- 11059923 TI - Spontaneous regression of hepatocellular carcinoma. PMID- 11059924 TI - A long way toward understanding the pathogenesis of functional dyspepsia, but we should go forward with the accumulation of clinical evidence. PMID- 11059925 TI - Portal-systemic encephalopathy in non-cirrhotic patients: classification of clinical types, diagnosis and treatment. AB - Hepatic encephalopathy is suspected in non-cirrhotic cases of encephalopathy because the symptoms are accompanied by hyperammonaemia. However, the cause of the large portal-systemic shunt formation observed in these cases is not clear, as cirrhosis and portal hypertension are absent. The frequency of such cases reported in the literature is increasing with progress and spread of abdominal imaging diagnostic techniques. Some cases have been misdiagnosed as psychiatric diseases (dementia, depression and others) and consequently patients have been hospitalized in psychiatric institutions or geriatric facilities. Some paediatric cases have also been misdiagnosed. Therefore, the importance of accurate diagnosis of this disease should be strongly emphasized. Some paediatric cases have also been misdiagnosed. When psychoneurological symptoms are suggestive of hepatic encephalopathy but objective and subjective symptoms or abnormal values of liver function tests are not sufficiently indicative of liver cirrhosis, portal-systemic encephalopathy should be suspected. Abnormal angiograms of the portal vein, superior mesenteric vein or splenic vein are conclusive evidence of portal-systemic encephalopathy. Transrectal portal scintigraphy also provides information useful for detection of shunts and a quantitative estimation of shunt index. We classified the disease into five types based on whether the shunt is formed inside or outside the liver. Type I (intrahepatic type) designates cases in which shunts are located between the portal and systemic veins. Type II designates a type of intra/extrahepatic shunt that originates from the umbilical part of the portal vein and serpentines in the liver, then leaves the liver. Type III (extrahepatic type) occurs most frequently. Type IV (extrahepatic) is accompanied by shunts similar to those in type III, but hepatic pathology presents as idiopathic portal hypertension. Type V (extrahepatic) represents the congenital absence of the portal vein, where the superior mesenteric vein joins the intrahepatic inferior vena cava or the left renal vein. The prevalence of each type in our country was examined by a nationwide investigation. In addition to the conventional diet or drug treatments, obliteration by less invasive interventional radiology using a metallic coil and ethanol has recently been used more frequently than surgical occlusion of shunts. Shunt-preserving disconnection of portal and systemic circulation and partial splenic artery embolization are also performed. International investigation of the disease status and establishment of diagnostic and therapeutic methods for the disease are awaited and investigation of long-term prognosis after therapy is also necessary. PMID- 11059926 TI - Magnetic resonance imaging (MRI) and diseases of the liver and biliary tract. Part 1. Basic principles, MRI in the assessment of diffuse and focal hepatic disease. AB - Magnetic resonance imaging (MRI) relies on the physical properties of unpaired protons in tissues to generate images. Unpaired protons behave like tiny bar magnets and will align themselves in a magnetic field. Radiofrequency pulses will excite these aligned protons to higher energy states. As they return to their original state, they will release this energy as radio waves. The frequency of the radio waves depends on the local magnetic field and by varying this over a subject, it is possible to build the images we are familiar with. In general, MRI has not been sufficiently sensitive or specific in the assessment of diffuse liver disease for clinical use. However, because of the specific characteristics of fat and iron, it may be useful in the assessment of hepatic steatosis and iron overload. Magnetic resonance imaging is useful in the assessment of focal liver disease, particularly in conjunction with contrast agents. Haemangiomas have a characteristic bright appearance on T2 weighted images because of the slow flowing blood in dilated sinusoids. Focal nodular hyperplasia (FNH) has a homogenous appearance, and enhances early in the arterial phase after gadolinium injection, while the central scar typically enhances late. Hepatic adenomas have a more heterogenous appearance and also enhance in the arterial phase, but less briskly than FNH. Hepatocellular carcinoma is similar to an adenoma, but typically occurs in a cirrhotic liver and has earlier washout of contrast. The appearance of metastases depends on the underlying primary malignancy. Overall, MRI appears more sensitive and specific than computed tomography with contrast for the detection and evaluation of malignant lesions. PMID- 11059927 TI - Magnetic resonance imaging and diseases of the liver and biliary tract. Part 2. Magnetic resonance cholangiography and angiography and conclusions. AB - Magnetic resonance cholangiography (MRC) relies on the strong T2 signal from stationary liquids, in this case bile, to generate images. No contrast agents are required, and the failure rate and risk of serious complications is lower than with endoscopic retrograde cholangiopancreatography (ERCP). Data from MRC can be summated to produce an image much like the cholangiogram obtained by using ERCP. In addition, MRC and conventional MRI can provide information about the biliary and other anatomy above and below a biliary obstruction. This provides information for therapeutic intervention that is probably most useful for hilar and intrahepatic biliary obstruction. Magnetic resonance cholangiography appears to be similar to ERCP with respect to sensitivity and specificity in detecting lesions causing biliary obstruction, and in the diagnosis of choledocholithiasis. It is also suited to the assessment of biliary anatomy (including the assessment of surgical bile-duct injuries) and intrahepatic biliary pathology. However, ERCP can be therapeutic as well as diagnostic, and MRC should be limited to situations where intervention is unlikely, where intrahepatic or hilar pathology is suspected, to delineate the biliary anatomy prior to other interventions, or after failed or inadequate ERCP. Magnetic resonance angiography (MRA) relies on the properties of flowing liquids to generate images. It is particularly suited to assessment of the hepatic vasculature and appears as good as conventional angiography. It has been shown to be useful in delineating vascular anatomy prior to liver transplantation or insertion of a transjugular intrahepatic portasystemic shunt. Magnetic resonance angiography may also be useful in predicting subsequent variceal haemorrhage in patients with oesophageal varices. PMID- 11059928 TI - Inhibition of mitogen-induced murine lymphocyte proliferation by Helicobacter pylori cell-free extract. AB - BACKGROUND AND AIMS: Previous studies have shown that lysates of Helicobacter pylori inhibit mitogen-induced proliferation of human peripheral blood mononuclear cells. The objective of the present study was to determine whether H. pylori cell-free extract (HPCE) has similar effects on murine lymphoid cells and could, therefore, be used to further delineate the mechanisms of alteration of lymphocyte function by H. pylori. METHODS: The HPCE was prepared from a H. pylori reference strain and from five clinical strains with varying status of cagA and vacA. Mouse splenic and mesenteric lymph node cells were cultured in microwell plates in the presence or absence of varying concentrations of HPCE (0.625-12.5 microg/mL). T cell mitogens were added into the culture 2 h later and the cells were cultured at 37 degrees C in 5% CO2 for a further 72 h. Cell proliferation was determined by a non-radioactive rapid dye assay and the percentage inhibition caused by HPCE was calculated. RESULTS: Pre-exposure to HPCE significantly inhibited concanavalin A-induced proliferation of murine spleen and mesenteric lymph node cells (up to 100% inhibition; P < or = 0.01). The HPCE also inhibited lymphocyte proliferation stimulated by mitogens phorbol-myristate-acetate and ionomycin and by the anti-CD3epsilon monoclonal antibody (P < or = 0.05). The inhibition was dose-dependent, but independent of the presence of virulence genes cagA or vacA. Treatment of HPCE at 80 degrees C for 30 min, but not at 55 degrees C for 60 min, completely abolished its inhibitory action. The HPCE, pretreated with pronase E, proteinase K, trypsin, acid or alkali also completely lost its inhibitory effect (P < or = 0.01), while in contrast, treatment with carboxypeptidase and leucine aminopeptidase had no effect. CONCLUSION: Helicobacter pylori produces heat-labile proteins or peptides that suppress T cell mitogen-induced proliferation of murine lymphoid cells in a similar manner to that observed with human peripheral blood mononuclear cells. The mouse cell culture system can, therefore, be used as a model to further study the mechanisms of action and antigen specificity of these immunomodulatory factors. PMID- 11059929 TI - Pooled analysis of Helicobacter pylori eradication regimes in Asia. AB - BACKGROUND AND AIMS: Differences in Helicobacter pylori strains and their resistance to antimicrobials between Asian and Western countries may affect the success in eradicating this bacterium. Our objective was to systematically review the regimens that have been tested in Asia. METHODS: Data on anti-H. pylori therapies reported from Asia in a large number of publications identified up to December 1998 were pooled into a few groups based on the combination of drugs used. A comparison of different groups was made by calculating the pooled eradication rates. RESULTS: Seventy-three studies with 134 treatment arms were reviewed. Pooled eradication rates of dual, triple and quadruple therapies were 61.0, 86.5 and 93.4%, respectively. Proton pump inhibitor (PPI)-based combinations were more widely used and effective, with overall eradication rates of 90.7% in triple therapy and 93.4% in quadruple therapy. Bismuth combined with tetracycline and metronidazole also showed a high eradication rate of 92.0%. CONCLUSIONS: Proton pump inhibitor-based triple therapy with either clarithromycin, amoxycillin or metronidazole was one of the most commonly used and effective anti-H. pylori triple therapy regimens in Asia. The classical triple therapy with a bismuth, tetracycline and metronidazole combination has a similar efficacy. The results of anti-H. pylori treatment in Asia are not different from those in Western countries. PMID- 11059930 TI - Effect of acute and long-term oral tobacco use on oesophageal motility. AB - BACKGROUND AND AIMS: Nicotine administration is known to decrease lower oesophageal sphincter (LOS) pressure. Although a few studies have assessed the effect of tobacco on the LOS, the effect of acute and long-term oral tobacco use on oesophageal motility is not known. The study was designed to investigate the effect of acute and long-term oral tobacco use on LOS and distal oesophageal motility. METHODS: Thirty-six healthy men (aged 18-65 years, median 34 years; 18 oral tobacco users, 18 non-tobacco users) underwent oesophageal manometry using a water-perfusion system. After baseline manometry, tobacco users were asked to keep 0.5 g tobacco in their mouth for 10 min; non-users of tobacco were kept in quiet surroundings for a similar period. Manometry was then repeated. RESULTS: The LOS basal pressures were similar in tobacco users and non-tobacco users (mean +/- SD 15.4 +/- 6.3 vs 13.4 +/- 5.3 mmHg). In the distal oesophageal body, the velocity (4.4 +/- 3.1 vs 4.9 +/- 2.6 cm/s), amplitude (92.7 +/- 38.3 vs 84.8 +/- 33.2 mmHg) and duration of contraction (2.1 +/- 0.7 vs 1.7 +/- 0.9 s) were similar in tobacco users and non-users. Acute tobacco use did not affect these parameters. The numbers of abnormal waves (triple peaks and non-transmitted contractions) were also similar in the two groups. CONCLUSION: Oral tobacco use does not appear to affect LOS pressures and distal oesophageal motility acutely or in the long term. PMID- 11059931 TI - Real-time ultrasonographic assessment of antroduodenal motility after ingestion of solid and liquid meals by patients with functional dyspepsia. AB - BACKGROUND AND AIMS: Although antroduodenal motility has usually been studied by using manometric or scintigraphic methods, ultrasonography is an established, non invasive method to evaluate duodenogastric motility. We used ultrasonography to evaluate gastric motility in patients with functional dyspepsia. METHODS: Sixty four patients with functional dyspepsia and 36 asymptomatic healthy subjects were given liquid and solid test meals. We investigated the gastric emptying rate, motility index, and duodenogastric reflux for the liquid meal and gastric emptying time, half-emptying time, and motility index for the solid meal. RESULTS: After the liquid meal, the gastric emptying rate and motility index were significantly lower and the duodenogastric reflux was significantly higher in functional dyspepsia patients than in healthy subjects. After the solid meal, gastric emptying time, half-emptying time and the motility index were significantly lower in the patients than in the healthy subjects. Delayed gastric emptying of both meals occurred in only 20.3% of patients. Delayed emptying of the liquid or solid meal occurred in 62.5% of patients. In both groups, gastric emptying time of the solid meal was positively correlated with the motility index at 15 min post-ingestion. CONCLUSION: In functional dyspepsia patients, delayed gastric emptying of a solid meal was related to antral hypomotility during the early postprandial phase. Ultrasonographic assessment of gastric motility in both liquid and solid meals may provide a better understanding of the pathogenesis of functional dyspepsia. PMID- 11059932 TI - Mortality association of enhanced CD44v6 expression is not mediated through occult lymphatic spread in stage II colorectal cancer. AB - BACKGROUND AND AIMS: In the absence of other metastatic disease, the presence of lymph node metastasis remains the most important determinant of survival in colorectal cancer (CRC). Cluster designation 44 variant 6 (CD44v6) over expression is associated with worse outcome in all stages of CRC. The CD44v6 is believed to confer metastatic potential through its facilitation of migration, extravasation and proliferation, although the specific means by which it conveys an adverse prognosis in CRC is unknown. The aim of the present study was to determine if CD44v6 over-expression in Stage II CRC subjects was associated with the presence of lymph node micrometastases. METHODS: We assessed tumour CD44v6 expression in 43 randomly sampled subjects who had resections for Stage II CRC between 1984 and 1991 by using immunohistochemistry. Micrometastases were sought in corresponding lymph node (LN) sections using keratin immunohistochemistry. RESULTS: There was a statistical trend between tumour CD44v6 over-expression and mortality (P = 0.09) and a significant relationship between LN cytokeratins and mortality (P = 0.01). There was no association between the detection of LN cytokeratins and tumour CD44v6 over-expression. CONCLUSION: We conclude that the adverse survival effect of CD44v6 over-expression is not mediated though lymphatic spread and postulate that it may therefore facilitate haematogenous metastasis. PMID- 11059933 TI - Adult coeliac disease: prevalence and clinical significance. AB - BACKGROUND AND AIMS: Although coeliac disease is a common condition, the role of population screening is not clear. The aim of this study was to determine the prevalence and clinical significance of coeliac disease in the adult population of Christchurch, New Zealand. METHODS: A total of 1064 adults randomly selected from the 1996 Christchurch electoral rolls were enlisted. The subjects were screened for coeliac disease using the anti-endomysial antibody test (EMA), and all those with positive tests were reviewed and underwent a small bowel biopsy. RESULTS: Twelve of the 1064 persons tested (1.1%) were EMA positive and all had small bowel biopsy histology consistent with coeliac disease. Two of the 12 subjects were previously known to be EMA positive although neither had a small bowel biopsy. One additional subject with known and treated coeliac disease was also enrolled but was EMA negative. Thus, the overall prevalence of coeliac disease was 13 of 1064 subjects (1.2%, or 1:82), 10 of whom were newly diagnosed (0.9%, or 1:106) and three were previously known or suspected to have coeliac disease (0.3%, or 1:355). The prevalence in both sexes was similar. Nine of the 12 EMA-positive coeliac disease subjects identified by the use of screening reported symptoms, of which tiredness and lethargy were the most common. The subjects were of normal stature, although females tended to be lean. None of the subjects were anaemic, but four were iron deficient and four folate deficient. Five of the 12 had sustained bone fractures. Bone mineral density was reduced in males but not in females. CONCLUSIONS: The prevalence of coeliac disease in the adult population of Christchurch, New Zealand, is 1.2%. Unrecognized coeliac disease which was detected by population screening was three-fold more common than proven or suspected coeliac disease. Population screening may identify subjects who could benefit from treatment. PMID- 11059934 TI - Incidence and prevalence of ulcerative colitis in the Songpa-Kangdong District, Seoul, Korea, 1986-1997. AB - BACKGROUND AND AIMS: Ulcerative colitis (UC) is regarded as a rare disease in developing countries, but accurate data are generally lacking. We performed the present study to evaluate the incidence and prevalence of UC in Korea. METHODS: A retrospective study was performed from 1986 to 1997 in the Songpa-Kangdong district of Seoul, Korea. To recruit UC patients as completely as possible, multiple information sources including all medical facilities in the study area and three referral centres located nearby, but outside the study area were used. The incidence and prevalence rates were adjusted using the 1997 Korean population statistics. RESULTS: During the study period, a total of 94 incident cases were identified, for an adjusted mean annual incidence rate of 0.68 per 100,000 inhabitants. On 31 December 1997, 91 patients with UC lived in the study area, giving an adjusted prevalence rate of 7.57 per 100,000 inhabitants. By using the Poisson regression analysis, the annual incidence rate increased significantly from 0.20 per 100,000 inhabitants in 1986-1988 to 1.23 per 100,000 inhabitants in 1995-1997 (P < 0.005). Patient age at diagnosis, the interval from onset of symptoms to diagnosis, and the disease extent at diagnosis were fairly constant throughout the study period. CONCLUSIONS: The incidence and prevalence of UC in our study area are still low compared with those of Western countries, but the incidence rate is steadily increasing. PMID- 11059935 TI - Detection of hepatitis A viral RNA in sera of patients with acute hepatitis A. AB - BACKGROUND AND AIMS: The Detection of hepatitis A virus (HAV) is important for diagnosis and epidemiological studies of hepatitis A. The polymerase chain reaction (PCR) technique is a sensitive test to detect HAV-RNA in specimens. The aims of the present study were to clarify the detection rate of serum HAV-RNA by PCR and the natural history of HAV viraemia, and to determine the correlation between viraemia and the clinical characteristics in patients with acute hepatitis A. METHODS: Hepatitis A virus RNA was tested in 74 serum samples which were serially collected from 27 patients with acute hepatitis A. A nested reverse transcription (RT)-PCR for HAV-RNA was performed with primer sets located at the VP1 region of the HAV genome and the PCR products were electrophoresed on a 1.5% agarose gel. RESULTS: Hepatitis A virus RNA was found in 18 of 27 (67%) patients with hepatitis A. There were no significant differences between groups positive and negative for HAV-RNA in clinical and laboratory data, except the time interval between clinical onset and initial serum sampling for RT-PCR (10 +/- 6 vs 19 +/- 14 days) and the alanine aminotransferase (ALT) level at initial serum sampling for RT-PCR (1436 +/- 1416 vs 518 +/- 432 IU/L). The mean duration of HAV viraemia was 30 +/- 19 days (range, 5-59 days). The duration of HAV viraemia and duration of abnormal ALT levels from clinical onset were positively correlated (r = 0.685, P = 0.007). CONCLUSION: In conclusion, HAV-RNA RT-PCR is a useful tool to detect HAV viraemia and to study the molecular epidemiology of HAV infection. PMID- 11059936 TI - Sequence-motif analysis of 5'-untranslated region of GB virus-C in Japanese patients. AB - BACKGROUND: GB Virus C (GBV-C) is considered to belong to the Flaviviridae; however, the structures of the N-terminal end of its putative polyprotein are not well known. The internal ribosomal entry site (IRES) at the 5'-untranslated region of GBV-C and an initiating codon at nucleotides (nt) 552-554 have been proposed. We investigated the validity of this proposal. METHODS: The 5' untranslated region of GBV-C was amplified from serum samples of 17 Japanese patients. Polymerase chain reaction-amplified products were directly sequenced and the obtained sequences were analysed by comparing them with the IRES structure of other viruses. RESULTS: Fifteen of the 17 (88%) GBV-C strains in our patients were classified as being Asian type. The box-A-like sequence (UUUC) and box-B-like sequence (AUCAUGG) observed in the IRES of picornaviruses were highly conserved in all the strains. Based on pair-wise comparisons with the multiple alignment data, overall sequence divergence for the 5'-terminus was 2.9-12%. When compared with the proposed secondary structure of the IRES model, the sequence divergences of the Asian-type GBV-C were higher at the regions of loop structures and lower at the regions of double-stranded RNA. The AUG codons, except for the one located at nt 552-554, produced truncated polyproteins or were not in-frame with the putative protein. CONCLUSIONS: Our examination of the sequence motif of GBV-C supports the proposal that the GBV-C has common structural motifs for IRES at its 5'-untranslated region and the AUG codon at nt 552-554 may be an initiating codon. PMID- 11059937 TI - Hepatitis C virus clearance is prominent in women in an endemic area. AB - BACKGROUND: The clinical and virological backgrounds of cases with previous hepatitis C virus (HCV) infection (positive for HCV antibody (anti-HCV) and HCV RNA negative) in an HCV endemic area were examined to identify factors related to the clearance of HCV. METHODS: The study population comprised 3117 inhabitants, 1037 male and 2080 female, from an HCV endemic area. Hepatitis C virus antibody was detected by a passive haemagglutination test. The HCV-RNA and the HCV genotype were detected by using the polymerase chain reaction method. The HCV serotype was determined by enzyme immunoassay by using the peptides of the core region. RESULTS: Twenty-two per cent of the inhabitants were positive for anti HCV, with males and the elderly having a significantly higher antibody titre (P < 0.01) than youths and females. Hepatitis C virus-RNA was detected in 78% of the HCV antibody-positive cases. The rate of HCV-RNA positivity was significantly higher in males than in females (P < 0.01). No relationship was found between HCV RNA positivity and age. The HCV genotype 1b was the predominant genotype among the HCV-RNA-positive cases. Mixed genotypes (1b + 2a) were observed in 12% of cases, primarily in elderly males and females. In cases with previous HCV infection, serotype 1 was the most common serotype, and there appeared to be no relationship between the distribution of HCV serotypes and age and gender. There was a female predominance with regard to previous HCV infection, but not to being HCV carriers (P < 0.01). CONCLUSIONS: Gender, not HCV genotype, is the primary factor influencing HCV clearance. PMID- 11059938 TI - Acute and chronic effects of isosorbide-5-mononitrate administration on effective renal plasma flow and the renin-aldosterone system in cirrhotic patients. AB - BACKGROUND AND AIMS: Isosorbide-5-mononitrate (ISMO) has been shown to be effective in reducing the risk of variceal bleeding in patients with cirrhosis. However, recent studies have suggested that this drug compromises renal function. The present study was conducted to assess the acute and chronic effects of ISMO on effective renal plasma flow (ERPF) and the renin-aldosterone profile in cirrhotic patients. METHODS: Fifteen cirrhotic patients were included in the present study. The mean arterial pressure (MAP), heart rate (HR), serum renin concentration (SR), ERPF and plasma aldosterone concentration (PA) were checked before ISMO treatment (baseline study), after a single oral dose of 20 mg ISMO (acute effect study) and after 3 weeks of ISMO treatment (chronic effect study). RESULTS: Our data showed that the oral administration of a single dose (20 mg) of ISMO to cirrhotic patients was associated with significant decreases in ERPF (from 405.18 to 369.06 mL/min) and MAP (from 93.26 to 86.40 mmHg), and increases in HR (from 65.53 to 70.06 beats/min), SR (from 24.15 to 54.41 pg/mL), and PA (from 105.1 to 148.7 pg/mL). However, no significant changes were observed in HR, MAP, PA, SR, or ERPF after 3 weeks of ISMO treatment when compared with the baseline study. CONCLUSIONS: The administration of ISMO causes a decrease in ERPF in cirrhotic patients and its use in patients with renal impairment should be considered cautiously. PMID- 11059939 TI - Telomerase activity in peripheral blood for diagnosis of hepatoma. AB - BACKGROUND: Telomerase activity may be used as a molecular marker for the detection of circulating hepatoma cells in blood of patients with hepatoma. METHODS: Telomerase activity in peripheral blood from hepatocellular carcinoma (HCC) patients was assessed by using a highly sensitive and non-radioisotope telomerase polymerase chain reaction (PCR) ELISA. Initially, tissue telomerase activity was measured in the hepatoma and non-tumour portions by using PCR ELISA within the same specimen, to compare its sensitivity with the conventional telomeric repeat amplification protocol (TRAP) method. Second, telomerase activity was measured in the peripheral blood obtained from patients with HCC, patients with chronic liver disease and in healthy controls. RESULTS: Of the 17 HCC patients, telomerase activity was found to be positive in 14 (82%) by using TRAP and 15 (88%) by using PCR ELISA, indicating that PCR ELISA is a reliable tool for the measurement of telomerase activity. By using the Telomerase PCR ELISA assay, telomerase activities in the peripheral blood of 20 HCC patients was 1.65 +/- 0.78 units. This was significantly greater than the results obtained for 20 chronic liver disease patients (0.43 +/- 0.36 units) and 20 healthy controls (0.39 +/- 0.14 units; P < 0.0001).When the arbitrary cut-off level was set at 0.7 units (maximum value of healthy controls + 0.1), the positive frequency of telomerase activity was 25% for chronic liver disease and 80% for HCC patients (sensitivity 80%, specificity 75%). Among the HCC patients, high telomerase activity in the peripheral blood was shown at stage III HCC with vascular invasion (2.10 +/- 0.62 units, n = 9). This was significantly higher than patients at stage II of HCC (1.28 +/- 0.72 units, n = 11, without vascular invasion; P = 0.012). CONCLUSION: These results suggest that peripheral blood telomerase activity, which may reflect haematogenous micrometastasis, is potentially a practical diagnostic/predictive marker of HCC. PMID- 11059940 TI - Human biliary beta-glucuronidase activity before and after relief of bile duct obstruction: is it the major role in the formation of pigment gallstones? AB - BACKGROUND AND AIMS: The bacterial beta-glucuronidase (bBG) can deconjugate conjugated bilirubin to form calcium bilirubinate gallstone. Yet, the role of the human biliary beta-glucuronidase (hBG) in the pathogenesis of pigment gallstone formation still remains unsolved. METHODS: Hepatic bile was collected from bile duct-obstructed patients on the day of, and 3 days after, biliary drainage. Patients were divided into pigment-stone (PS) group (n = 34) and stone-free (SF) group (n = 29). All patients of the PS group had the complication of cholangitis. The concentrations of bile contents and the activities of bBG and hBG were measured. RESULTS: The activities of hBG and bBG in bile obtained on the day of biliary drainage were higher in the PS group than in the SF group (activities corrected for bile salt concentration: hBG 128.7 +/- 340.0 vs 13.1 +/- 25.0 U/mmol; bBG 12.5 +/- 22.2 vs 4.6 +/- 7.7 U/mmol, P < 0.05). This difference disappeared after biliary drainage. The changes of enzyme activity in the bile of the SF group were unremarkable before and after biliary drainage. The mean concentrations of bile pigments and free calcium in the PS group were lower than those in the SF group. CONCLUSIONS: An increase in the activity of hBG may be a secondary response, developed after bile duct inflammation because it was elevated only when the bile duct obstruction was associated with infection. PMID- 11059941 TI - Images of interest. Gastrointestinal: Behcet's disease. PMID- 11059942 TI - Images of interest. Hepatobiliary and pancreatic: a man with a dull abdominal pain. PMID- 11059943 TI - Spontaneous regression of hepatocellular carcinoma and review of literature. AB - A 68-year-old man presented with multiple hepatocellular carcinoma, which was considered to be unresectable at the first admission in January 1994. Pathological diagnosis was made by biopsy of the one lesion among them. From January 1994 to December 1997, 10 transarterial chemoembolizations and six percutaneous ethanol injection therapies were performed on the tumours in the cirrhotic liver. In February 1998 the tumour situated in the right lobe began to increase in size. The maximum tumour diameter was 6.3 cm measured by computed tomography (CT). In the beginning of May 1998 moderate ascites was present and mild hepatic encephalopathy was noticed. The patient was in the terminal stage of hepatocellular carcinoma and no further treatment was possible at that time. However, serum alpha-fetoprotein and protein induced by vitamin K absence or antagonist II dramatically decreased in June 1998. The CT scan also showed that the tumour had completely regressed without specific treatment. In February 1999 a new biopsy-proven hepatocellular carcinoma, 2 cm in diameter, developed in the lateral segment of the liver. It was well treated by percutaneous ethanol injection therapy. The patient was alive in good condition without any symptoms or tumour recurrence in June 1999. It was concluded that a rare case of spontaneous regression of hepatocellular carcinoma had occurred. PMID- 11059944 TI - Diet and asthma. AB - The role of food intolerance in asthma is well recognized, and where food avoidance measures are instituted considerable improvement in asthma symptoms and in reduction in drug therapy and hospital admissions can result. These benefits may have a greater impact in those patients with greater symptoms. However, the promise of such benefits should not result in an approach which ignores inhaled drug therapy, or in a dietary regime which is inappropriate in the face of mild symptoms. Whilst sub-optimal intake of dietary nutrients is also a recently recognized potential risk factor for asthma, available data are insufficient to implicate any as casual. A number of studies have sought to establish the role of the antioxidant vitamins, A, C and E and selenium, yet others of the elements sodium and magnesium. Sub-optimal nutrient intake may enhance asthmatic inflammation, consequently contributing to bronchial hyperreactivity. Prospective studies of supplementation therapy are needed to confirm this. PMID- 11059945 TI - Effects of naturally-occurring acid fog on inflammatory mediators in airway and pulmonary functions in asthmatic patients. AB - Floating fog occurs every summer in Kushiro City in Japan, and the annual average of fog water pH in the past 4 years has been under 5.0. We previously reported that epidemiologically fog was the most important positive factor contributing to increased hospital visits of asthmatic patients compared with other meteorological values and air pollutants. This study aimed to investigate the mechanism of the effects of naturally-occurring acid fog on asthmatic patients. We compared pulmonary functions and inflammatory mediators in induced sputum between the foggy (July 1995) and the non-foggy (May 1996) season, and assessed airway responsiveness to hypo-osmolar aerosol. Forty-four out of 118 asthmatic patients of Kushiro City residents participated, pulmonary function tests were completed in 36 patients, and sputum data were available in 26 patients in both seasons. Percent forced expiratory volume in 1 sec (FEV1) was significantly (P< 0.05) decreased, and % peak expiratory flow rate (PEFR) had a trend to decrease in the foggy season more than in the non-foggy, and sputum eosinophil cationic protein (ECP) and interleukin (IL)-8 were higher in the foggy season but not significantly. A moderate inverse correlation was revealed between sputum ECP and %PEFR in the foggy season (r= -0.55, P<0.005). Subjects were divided into two groups according to the best PEFR; one had >10% lower PEFR levels in the foggy season than in the non-foggy season (Group A, n = 7), the remainder did not (Group B, n = 19). In group A, sputum ECP was significantly increased (P< 0.01) in the foggy season, but there were no changes in IL-8 and prostaglandin D2. Ultrasonic nebulized distilled water provocation test revealed no differences between group A and B. These results suggested that eosinophilic inflammation rather than hypo-osmolar effect of fog might contribute to respiratory deterioration by inhalation of naturally-occurring acid fog. PMID- 11059946 TI - Chemoradiotherapy for advanced lymphoepithelioma-like carcinoma of the lung. AB - Lymphoepithelioma-like carcinoma (LELC) of the lung, an Epstein-Barr virus associated undifferentiated carcinoma, is a rare entity of pulmonary malignancy. It tends to affect young non-smoking Asians and is often resectable. However, little is known of the treatment of the even rarer locally advanced or metastatic cases. We report our experience of three Chinese patients with advanced LELC of the lung who were treated with combination-chemotherapy (5-fluorouracil, leucovorin, and cisplatin) and radiotherapy. The encouraging response of these patients supports the use of this regime in other patients. PMID- 11059947 TI - Fenoterol hydrobromide delivered via HFA-MDI or CFC-MDI in patients with asthma: a safety and efficacy comparison. AB - The main objective of the study was to compare the long-term safety and tolerability of fenoterol hydrobromide delivered using a metered-dose inhaler formulated with the alternative propellant, hydrofluoroalkane 134a (HFA-MDI), with delivery using the currently available chlorofluorocarbon MDI (CFC-MDI; Berotec 100). A further objective was to compare the efficacy of fenoterol HFA MDI with fenoterol CFC-MDI, using the pulmonary function parameters of forced expiratory volume in 1 sec (FEV1), forced vital capacity (FVC) and peak expiratory flow (PEF). Following a 2-week run-in phase, a 12-week, double-blind parallel group comparison was undertaken in 290 patients randomized on a 2:1 basis to two puffs of 100 microg fenoterol four times a day (HFA-MDI=197 patients; CFC-MDI=93 patients). A total of 236 patients in this multi-centre study completed the trial as planned. The overall incidence of adverse events (AEs) was similar in both groups (29.9% of HFA-MDI patients and 28% of CFC-MDI patients). Reports of respiratory disorder AEs were also comparable (21.8% HFA MDI; 22.6% CFCMDI). End of study laboratory tests, ECG, pulse, blood pressure and physical examination showed no significant differences from pre-study baselines in either group and both treatments appeared to be well tolerated. Pre-dose FEV1 measurements taken at the three clinic visits were constant and increase in FEV1 at 5 and 30 min post-dose demonstrated equivalent efficacy for the two formulations. No difference between the two groups was observed in PEF or in the use of rescue medication. We conclude from these findings that the long-term safety and efficacy profile of fenoterol HFA-MDI is comparable to that of the fenoterol CFC-MDI. PMID- 11059948 TI - Risk factors for community-acquired pneumonia diagnosed upon hospital admission. British Thoracic Society Pneumonia Study Group. AB - A case-control study of risk factors for community-acquired pneumonia in adults admitted to hospital is reported. Cases were surviving patients (n = 178) admitted to 14 hospitals in England. Controls were individuals (n = 385) randomly selected from the electoral registers of the areas served by the hospitals. The two groups were compared with regard to risk factors for pneumonia using a standardized postal questionnaire. Independent risk factors associated with cases in log-linear regression analysis were age, heart disease (as indicated by congestive heart failure and/or digitalis treatment), lifetime smoking history, chronic airway disease (chronic bronchitis and/or asthma), occupational dust exposure, pneumonia as a child, single marital status and unemployment. Corticosteroid and bronchodilator therapy were also independent risk factors in the log-linear regression analysis, but may reflect the severity of underlying lung disease for which these drugs were prescribed. These data suggest that cigarette smoking is the major avoidable risk factor for acute pneumonia in adults. PMID- 11059949 TI - Human leucocyte antigens (HLA) and trimellitic anhydride (TMA) immunological lung disease. AB - Occupational immunological lung disease, due to low molecular weight, reactive chemicals such as trimellitic anhydride (TMA), is an emerging health problem. If there were a marker that was highly predictive of the ability of the immune system to recognize TMA as an allergen, better prevention strategies could be employed with at risk individuals. The purpose of this study is to evaluate whether human leucocyte antigen (HLA) class specificity is associated with the development of late respiratory systemic syndrome (LRSS) or asthma due to immunological sensitivity to trimellitic anhydride (TMA). This is a case control study of 17 individuals with LRSS, 12 with asthma and 22 TMA similarly exposed individuals who did not develop LRSS or asthma. Comparing the sensitized individuals (LRSS or asthma) with the non-sensitized individuals (controls), we found no difference in frequency of any HLA antigen. In summary, the lack of association of HLA antigens with LRSS or asthma due to TMA suggests that these will not be useful markers to identify at risk individuals. PMID- 11059950 TI - Self-reported sleepiness while driving as a risk factor for traffic accidents in patients with obstructive sleep apnoea syndrome and in non-apnoeic snorers. AB - The obstructive sleep apnoea syndrome (OSAS) is a condition causing daytime sleepiness and has been related to an increased risk for traffic accidents. However, the evidence linking severity of OSAS to a higher rate of automobile crashes is based on limited data. The aims of this study were to study the traffic accident rate in the last 5 years in patients referred to our sleep clinic because of clinical suspicion of OSAS and to analyse variables related to an increased risk for traffic accidents. A series of 189 consecutive patients with a driving license referred for a sleep study because of OSAS clinical suspicion and a control group (CG) of 40 hospital staff workers who denied snoring, matched for age and sex with the study population, were studied. Patients underwent a full-night polysomnography and both patients and the CG completed a self-answered questionnaire. One hundred and twenty-two patients were diagnosed as OSAS and 67 patients as non-apnoeic snorers (NAS). The self-reported number of accidents was significantly higher in OSAS patients compared with CG. The self-reported number of times off the road was significantly higher in OSAS patients compared with NAS and with CG. Variables associated with an increased risk for traffic accidents were self-reported sleepiness while driving (OR 5, 95%CI 2.3-10.9), having quit driving because of sleepiness (OR 3, 95%CI 1.1-8.6) and being currently working (OR 2.8, 95%CI 1.1-7.7). We conclude that self reported sleepiness while driving is associated with an increased risk for traffic accidents in OSAS patients and in NAS. We suggest that this symptom can be used to alert patients and to give priority in the sleep clinic for study and treatment. PMID- 11059951 TI - Is asthma underestimated as a cause of sick leave? AB - Public interest needs to be focused on the economic burden of asthma on society because of the increasing prevalence of the condition. Asthma is common in individuals of working age and sick leave is an important health-economic issue. In the present study we looked at the prevalence of asthma in a sick leave register. Individuals on sick leave due to asthma, individuals on sick leave due to any other respiratory disorder or symptom and individuals on sick leave due to non-respiratory conditions were included in a questionnaire based study. Individuals in the register diagnosed with asthma could be classified as current asthmatics or possible asthmatics in respectively 94% and 99% of the cases. They were also ex-smokers to a greater extent than the other groups, which was more pronounced in males. However, individuals on sick leave due to 'any other respiratory disorder' could be classified as current asthmatics or possible asthmatics in respectively 19% and 30% of the cases. The corresponding figures in the group on sick leave due to 'other diagnoses' were 7% and 10%. Hence, there is evidence that asthma is an under-reported diagnosis and this must be taken into consideration when sick leave registers are used in health-economic studies. PMID- 11059952 TI - The availability of smoking cessation advice from health professionals--a census from one East London District. AB - A survey of 382 hospital inpatients and a survey of 500 adults attending a GP open access chest X-ray service showed that 18% and 25% respectively were current smokers. Sixty per cent of the inpatient smokers and three quarters of the community smokers expressed a wish to stop smoking, and 44% of the inpatient smokers and 62% of the community smokers reported having received advice from their primary care physician to stop smoking. However, when the community smokers were asked about more specific advice they had received to help them stop smoking, only 13% had received advice regarding nicotine replacement therapies and under 5% had been given the telephone number of a smoking cessation support service (Smokers Quitline). Use of nicotine replacement therapies nearly doubles the success rate for smoking cessation, and it is essential for all health professionals to be able to give specific advice as to how smokers may be able to quit. PMID- 11059953 TI - Exhaled nitric oxide partitioned into alveolar, lower airways and nasal contributions. AB - During the last year exhaled nitric oxide (NO) has been proposed as a marker of airway inflammation. More knowledge of the production and transfer of this molecule are needed in order for NO analysis to become a clinical tool. This was the aim of the study. Exhaled NO values from multiple flow rates were used to model alveolar NO, transfer rate and tissue concentration of NO in the airways. Three flows rates, 0.005, 0.1 and 0.51 sec(-1) were found to be optimal. The NO transfer rate of the airways was 9 +/- 2 ml sec(-1), the tissue source was 75 +/- 28 ppb and the alveolar fraction of NO was 2 +/- 1 ppb in 10 healthy subjects (mean +/- CI95%). In conclusion, we have shown that it is possible to get more information about the distribution of NO in the lungs and the airways than only a single value from one expiratory flow rate can give. Further studies will reveal if this airway modelling can be useful in disease of the respiratory system. PMID- 11059954 TI - Arterial and bronchoalveolar lavage fluid endothelin-1 concentration in asthma. AB - Endothelins are a family of peptide mediators that have a number of biological properties, including the ability to act as bronchoconstrictors and vasoconstrictors of isolated airways and vessels. Endothelin-1 (ET-1) is the more potent peptide compared with the other two peptides of the family. To examine a possible involvement of ET- 1 in the pathogenesis of asthma, we measured arterial ET-1 levels in 11 patients with atopic asthma during attack and during remission, and in 11 healthy control subjects. We also performed fiberoptic bonchoscopy and bronchoalveolar lavage (BAL) to measure ET-1 levels in the 11 asthmatic patients during remission, and in the 11 healthy control subjects. ET-1 concentrations in arterial blood and in BAL were measured by a radioimmunoassay method. Arterial ET 1 levels were very significantly higher in asthma attack (3.67 +/- 0.51 pg ml( 1)) and in asthma remission (2.65 +/- 10.62 pg ml(-1)) compared with those of the healthy controls (1.37 +/- 0.14 pg ml(-1)) (P < 0.001). Arterial ET-1 levels were also very significantly higher during asthma attack compared with those in asthma remission (P < 0.001). BAL ET-1 levels were significantly higher in asthma remission (0.73 +/- 0.53 pmol g(-1)) compared with those of the healthy controls (0.16 +/- 0.02 pmol g(-1)) (P < 0.05). No correlation was observed between arterial and BAL ET-1 levels, PaO2 and peak expiratory flow rate (PEFR). These data are consistent with the hypothesis that ET-1 contributes to the pathophysiology of asthma. However, it is likely that the true importance of this vasoconstrictor peptide will only be revealed by pharmacological studies with specific receptor antagonists. PMID- 11059955 TI - Is it possible to predict the success of non-invasive positive pressure ventilation in acute respiratory failure due to COPD? AB - There is now sufficient evidence that non-invasive positive pressure ventilation (NIPPV) in selected patients with severe hypercapnic acute respiratory failure due to chronic obstructive pulmonary disease (COPD) is more effective than pharmacological therapy alone. The aim of this study was to identify prognostic factors to predict the success of this technique. Fifty-nine consecutive patients with COPD admitted to a respiratory ward for 75 episodes of acute respiratory failure treated with NIPPV were analysed: success (77%) or failure (23%) were evaluated by survival and the need for endotracheal intubation. There were no significant differences in age, sex, cause of relapse and lung function tests between the two groups. Patients in whom NIPPV was unsuccessful were significantly underweight, had an higher Acute Physiology and Chronic Health Evaluation (APACHE) II score, and a lower serum level of albumin in comparison with those in whom NIPPV was successful. They demonstrated significantly greater abnormalities in pH and PaCO2 at baseline and after 2h of NIPPV. The logistic regression analysis demonstrated that, when all the variables were tested together, a high APACHE II score and a low albumin level continued to have a significant predictive effect. This analysis could predict the outcome in 82% of patients. In conclusion, our study suggests that low albumin serum levels and a high APACHE II score may be important indices in predicting the success of NIPPV. PMID- 11059956 TI - Questionnaire survey of male infertility in cystic fibrosis. AB - Most male cystic fibrosis (CF) patients are infertile due to obstructive azospermia but little is known about the best time to counsel patients on infertility. All male patients attending the Adult Nottingham CF unit were invited to complete an anonymous questionnaire on infertility. The response rate was 60%. The median age that the patients first became aware of male infertility was 17 years (range 13-24) but the preferred age of receiving this information was 14 years (range 8-16). Patients first learnt about male infertility from the CF team (six patients), parents (five), from written information (two) or unexpectedly (five). Five out of 18 patients had undergone seminal analysis at a median age of 26 years but 17/18 patients felt that this should be offered routinely. Our survey has shown that patients would like infertility discussions at a younger age and routine seminal analysis. PMID- 11059957 TI - Case reports of death during pregnancy in patients with cystic fibrosis--three out of four patients were colonized with Burkholderia cepacia. PMID- 11059958 TI - Cattle TB schemes: control or eradication--a critical reappraisal. PMID- 11059959 TI - Pulmonary fibrosis in association with human T cell lymphotropic virus type 1 (HTLV-1) infection. PMID- 11059960 TI - The use of biopsy to study airway inflammation. PMID- 11059961 TI - Designing bronchial biopsy studies. AB - Bronchial biopsy provides valuable information about the inflammatory processes in lung tissue, but optimal results are only achieved if the design of intervention studies is sufficiently rigorous. The parallel-group design has merit, but the cross-over design is statistically superior, providing the wash out period is effective. Heterogeneity of contributing pathologies in asthma patients results in large inter-patient variability which must be controlled for, for example by using strict inclusion criteria, which should ideally relate to the specific inflammatory marker being studied. The inclusion of a placebo group helps to quantify sample variability. The study must have sufficient statistical power to detect inter-group differences for each variable; appropriate adjustments should be made when multiple tests are used. Studies with larger patient numbers are best performed using a multi-centre design, with one centre analysing all tissue samples to reduce variability. Study duration depends on the type of investigation, but should ideally be short. Longer studies are necessary to evaluate chronic changes such as tissue remodelling. Changes in clinical status and cellular events may follow different time courses after intervention. Biopsy measurements are less reproducible than physiological tests, and diurnal variation in the number and function of inflammatory cells can further complicate measurement. The timing of clinical trial assessments needs to allow for these idiosyncrasies. Finally, a balance must be maintained between the risk, albeit small, and the benefit of performing bronchial biopsies. PMID- 11059962 TI - Biopsy markers of airway inflammation and remodelling. AB - Bronchial inflammation is a consistent feature of asthma and its chronicity probably determines disease progression. Clinical evaluation of drugs with potential disease-modifying activity requires measurement of their effects on the inflammatory and remodelling process using a variety of techniques including bronchial biopsy, and analysis of sputum, bronchoalveolar lavage, blood, urine and exhaled air. Markers of the key components of the inflammatory process, such as the number and activation of T-cells. the number of mast cells, cytokine and chemokine release or gene expression, and eosinophil and neutrophil recruitment, can be determined in biopsy samples. Biopsies also allow assessment of the integrity and structure of the airway epithelium, the thickness of the reticular basement membrane and the numbers and ultrastructure of contractile cells. These and other markers may allow differentiation between subtypes of asthma patient according to atopic status and will help to distinguish asthma from chronic obstructive pulmonary disease. Airway remodelling may be a consequence of chronic bronchial inflammation and is a characteristic of chronic asthma, particularly in severe asthma and when there is progressive decline in lung function. There are changes in the surface epithelium, reticular basement membrane, bronchial smooth muscle, blood vessels and mucous glands. Reliable markers of remodelling need to be identified to improve our ability to evaluate chronic asthma therapy. PMID- 11059963 TI - The anti-inflammatory profile of inhaled corticosteroids: biopsy studies in asthmatic patients. AB - The beneficial effects of inhaled corticosteroids in the treatment of asthma are well established. A potent topical anti-inflammatory action is assumed to underlie the therapeutic effect, given that these agents alter the number and function of a range of inflammatory cells and markers in airway biopsies. This activity profile is shown by all inhaled corticosteroids, in a variety of patient types and study designs. Thus, treatment with inhaled corticosteroids leads to consistent reductions in the number and activation of mast cells and eosinophils in biopsy specimens. Other relevant findings include reductions in T-lymphocytes, which contribute to chronic inflammation via the secretion of pro-inflammatory cytokines (some of which are responsible for eosinophil accumulation and activation). Inhaled corticosteroids may therefore act by down-regulating immunoreactivity, so reducing activation of T lymphocytes and (consequently) eosinophils. There is considerable interest in whether corticosteroids can inhibit or reverse some structural changes in the airways, including basement membrane thickening, collagen deposition and increased airway vascularity; it has been suggested that these changes may contribute towards airway hyperresponsiveness and irreversible airway obstruction. In summary, inhaled corticosteroids have a broad spectrum of anti-inflammatory activity in asthma patients, but the relationship between changes in clinical and immunopathological parameters, particularly in the long-term, requires further study. PMID- 11059964 TI - The effect of long-acting beta2-agonists on airway inflammation in asthmatic patients. AB - Early observations suggested that the inhibition by long-acting beta2-agonists (LABAs) of non-specific bronchial hyperresponsiveness following allergen challenge was unrelated to bronchodilation or functional antagonism and might be a reflection of anti-inflammatory activity. Investigation of the effect of LABAs on airway inflammatory responses has demonstrated an inhibition of eosinophil recruitment in allergen challenge studies. Nevertheless, results from biopsy and other studies suggest that the chronic inflammatory process in asthma patients is unaffected by these drugs. There is no evidence from biopsy studies that LABAs are pro-inflammatory or that they mask existing inflammation. The beneficial effects of LABAs in allergen challenge are probably mediated through stabilization of mast cells. Recent evidence suggests that LABAs may reduce numbers of neutrophils and their associated markers; this observation needs to be confirmed in future studies and its relevance to the treatment of asthma determined. PMID- 11059965 TI - The anti-inflammatory profile of inhaled corticosteroids combined with salmeterol in asthmatic patients. AB - Inhaled corticosteroid (ICS) therapy such as fluticasone propionate (FP) is effective in moderate-to-severe asthma, but for patients on ICS who still experience symptoms, treatment guidelines recommend either increasing the dose of ICS or adding a long-acting beta2-agonist such as salmeterol or formoterol. Several studies have now shown that adding salmeterol provides greater clinical benefit than increasing the dose of ICS, raising the question of whether salmeterol has an additive or complementary anti-inflammatory effect to that of ICS. Recent studies on bronchial biopsies and bronchoalveolar lavage from asthmatic patients treated with either salmeterol. FP or placebo in addition to low-dose ICS have demonstrated that addition of salmeterol produces independent or additional reductions in several pro-inflammatory cells, cytokines and cell adhesion molecules compared with FP. Such complementary anti-inflammatory effects may explain the improved control of asthma symptoms and exacerbations observed when salmeterol is added to low-dose ICS therapy, and may help to modify the long term sequelae of asthma. These findings also indicate, contrary to earlier speculation, that salmeterol does not have a pro-inflammatory effect or mask persistent airway inflammation. This review presents the results of recent studies and suggests possible mechanisms for the additional antiinflammatory effects of salmeterol. PMID- 11059966 TI - Relating inflammatory changes in asthma to clinical status. PMID- 11059967 TI - Independent autonomic modulation of sinus node and ventricular myocardium in healthy young men during sleep. AB - INTRODUCTION: The aim of this study was to investigate whether autonomic modulation of ventricular repolarization may spontaneously differ from that of the sinoatrial node. METHODS AND RESULTS: Onset of P waves, QRS complexes, and the apex and end of T waves were detected beat to beat in high-resolution ECGs from nine healthy young men during the night. There were time-dependent fluctuations in the QT/RR slopes of consecutive 5-minute segments that could not be explained by the mean RR cycle length of the respective segment. Because the variability found in QT intervals could not be explained by either possible effects of rate dependence or hysteresis, autonomic effects were obvious. Power spectral analysis was performed for consecutive 5-minute segments of PP and QT tachograms. In a given subject, trends in the time course of low-frequency (LF) and high-frequency (HF) power in PP and QT often were similar, but they were quite different at other times. The mean LF/HF ratio for QTend (0.75 +/- 0.1) was different from that of PP (1.8 +/- 0.2; P = 0.002), indicating differences in sympathovagal balance at the different anatomic sites. Furthermore, at a given mean heart rate, averaged QT intervals were different on a time scale of several minutes to hours. The QT/RR slope of 5-minute segments correlated significantly with the HF power of QT variability but not with that of PP variability, indicating effects of the autonomic nervous system on ventricular action potential restitution. CONCLUSION: These differences demonstrate that changes in sinus node automaticity are not necessarily indicative of the autonomic control of ventricular myocardium. PMID- 11059968 TI - Gender-related differences in modulation of heart rate in patients with congestive heart failure. AB - INTRODUCTION: The prognosis of women with congestive heart failure (CHF) is better than that for men, but the mechanisms underlying the female survival advantage are not well understood. CHF is characterized by profound abnormalities in cardiac autonomic control that contribute to progressive circulatory failure and influence survival. METHODS AND RESULTS: Time- and frequency-domain heart rate variability (HRV) indexes were obtained from 24-hour Holter recordings and compared to assess the role of gender in 131 men and 68 women with CHF (mean age 60 +/- 13.6 years, range 21 to 87; New York Heart Association Functional Class III [66%] and IV [34%]). Gender-related differences in HRV were observed only in the subset of patients with nonischemic heart failure (55 men and 39 women). Among the time-domain indexes, the SD of the RR intervals (76 +/- 5.3 msec vs 55.3 +/- 3.2 msec, P < 0.0001) and indexes denoting parasympathetic modulation, the percentage of RR intervals with >50 msec variation (4.0% +/- 1.0% vs 6.5% +/- 1.3%, P = 0.02), and the square root of mean squared differences of successive RR intervals (19.1 +/- 3.3 vs 28.4 +/- 3.8, P = 0.004) were higher in women. Among the frequency-domain indexes, the total power (7.5 +/- 0.13 ln-msec2 vs 8.3 +/- 0.14 ln-msec2, P = 0.0002), the ultralow-frequency power (7.2 +/- 0.11 ln-msec2 vs 8.0 +/- 0.14 In-msec2, P < 0.0001), the low-frequency power (3.8 +/- 0.25 ln msec2 vs 4.8 +/- 0.28 ln-msec2, P = 0.006), and the high-frequency power (3.8 +/- 0.24 ln-msec2, vs 4.6 +/- 0.26 ln-msec2, P = 0.003) were greater in women than in men. CONCLUSION: Women with nonischemic CHF have an attenuated sympathetic activation and parasympathetic withdrawal compared with men. Gender-based differences in autonomic responses in the setting of CHF may be related to the female survival advantage. PMID- 11059969 TI - Circadian variation of vasovagal syncope. AB - INTRODUCTION: Circadian patterns have been demonstrated for several cardiovascular catastrophes. Chronobiologic factors play a role in the emergence of vasovagal syncope (VVS); however, diurnal variation of syncopal episodes in VVS has not been reported previously. METHODS AND RESULTS: We assessed daily distribution of the time of syncopal episodes in VVS. Time of syncope could be determined in 80 episodes in 54 patients (32 men and 22 women; mean age 37 years, range 12 to 67). Patients who were prescribed beta blockers or vasodilators, and who had syncopes related to alcohol intake, were excluded from the study. Head-up tilt testing was performed in 53 patients. The distribution of the episodes of VVS in 3-hour intervals differed significantly from uniform occurrence (P < 0.0001), with a peak frequency between 6 A.M. and noon (67.5% of total episodes). In patients who had experienced the initial syncope in the morning, most (78%) of the next syncopal episodes also occurred in the morning. There was no significant correlation between the time of last syncopes and tilt testing results. CONCLUSION: We demonstrated a prominent circadian variation in the frequency of VVS, with a peak in the morning. Recognition of the daily distribution of VVS is useful for patient education and therapeutic strategy. PMID- 11059970 TI - Long-term effects of biatrial synchronous pacing to prevent drug-refractory atrial tachyarrhythmia: a nine-year experience. AB - INTRODUCTION: Results of previous studies suggest that atrial resynchronization by multisite atrial pacing may contribute to prevention of recurrences in patients with drug-refractory atrial tachyarrhythmias and significant intra atrial conduction delay. METHODS AND RESULTS: To verify this hypothesis, a prospective noncontrolled study of 86 patients (mean age 66 +/- 10 years) was conducted in a single center between January 1989 and February 1998. Inclusion criteria were P wave duration > or = 120 msec with interatrial conduction time > or = 100 msec, and history of multiple recurrences of atrial tachyarrhythmias (mean 7 +/- 4.8 episodes) evolving in a persistent mode for at least 6 months despite optimized drug treatment (mean 2.7 +/- 1.8 drugs/patient). Patients were chronically implanted with a pacing system that ensured permanent biatrial pacing using two atrial leads, one placed in the high right atrium and the other one into the mid or the distal part of the coronary sinus. P wave duration decreased from a mean value of 187 +/- 29 msec before implant to 106 +/- 14 msec (P < 0.0001) under biatrial pacing. After a 33-month mean follow-up (range 6 to 109), 55 patients (64%) remained in sinus rhythm, including 28 patients (32.6%) without any documented recurrence and 27 patients with one or more recurrences in a paroxysmal or in a persistent form. In these 55 patients, drug treatment was significantly reduced in relation to the preimplantation period (1.4 +/- 0.6 vs 1.7 +/- 0.5 drugs/patient; P = 0.011). The other 31 patients went into chronic atrial arrhythmia after a mean period of 26 months. The only predictive factor of positive response was a spontaneous P wave duration < 160 msec at baseline. CONCLUSION: The results are consistent with a preventive effect of permanent biatrial pacing on recurrent and drug-refractory atrial arrhythmias associated with intra-atrial conduction delay. PMID- 11059971 TI - Is there a role for biatrial pacing in the prevention of atrial arrhythmias? PMID- 11059972 TI - Radiofrequency ablation of idiopathic left ventricular tachycardia at the site of earliest activation as determined by noncontact mapping. AB - INTRODUCTION: The most effective method for guiding radiofrequency (RF) ablation of idiopathic left ventricular tachycardia (ILVT) has yet to be determined. We investigated the use of noncontact mapping in five patients with this condition. METHODS AND RESULTS: The multielectrode array was positioned in the left ventricular apex via the retrograde approach. Isopotential color maps of ILVT were examined to determine the site of earliest endocardial activation. The ablation catheter was steered to the target site using the locator signal. Pace mapping was performed and contact electrograms examined for diastolic potentials. RF energy was applied to the target site. Sustained ventricular tachycardia was induced in 2 patients and nonsustained ventricular tachycardia in 3. The site of earliest activation was at the apical septum in 3, the inferior apex in 1, and the base of the inferior wall in 1. Mean timing was 21 +/- 10 msec before onset of the surface QRS. Diastolic activity was visualized with noncontact mapping at the base of the septum in 1 patient. A Purkinje potential was seen at the ablation site in only 1 patient. No diastolic activity was seen in the remaining 3 patients. Tachycardia was successfully terminated in all 5 patients with a median of four RF applications. No patient suffered a recurrence after 9.6 +/- 4.7 months of follow-up. CONCLUSION: By identifying the precise site of earliest activation during ILVT, noncontact mapping has been shown to be an effective and safe method for guiding RF ablation. PMID- 11059973 TI - Catheter ablation of idiopathic left ventricular tachycardia: use of new mapping technologies--when and why. PMID- 11059974 TI - Dual chamber arrhythmia detection in the implantable cardioverter defibrillator. AB - INTRODUCTION: Dual chamber implantable cardioverter defibrillator (ICD) technology extended ICD therapy to more than termination of hemodynamically unstable ventricular tachyarrhythmias. It created the basis for dual chamber arrhythmia management in which dependable detection is important for treatment and prevention of both ventricular and atrial arrhythmias. METHODS AND RESULTS: Dual chamber detection algorithms were investigated in two Medtronic dual chamber ICDs: the 7250 Jewel AF (33 patients) and the 7271 Gem DR (31 patients). Both ICDs use the same PR Logic algorithm to interpret tachycardia as ventricular tachycardia (VT), supraventricular tachycardia (SVT), or dual (VT+ SVT). The accuracy of dual chamber detection was studied in 310 of 1,367 spontaneously occurring tachycardias in which rate criterion only was not sufficient for arrhythmia diagnosis. In 78 episodes there was a double tachycardia, in 223 episodes SVT was detected in the VT or ventricular fibrillation zone, and in 9 episodes arrhythmia was detected outside the boundaries of the PR Logic functioning. In 100% of double tachycardias the VT was correctly diagnosed and received priority treatment. SVT was seen in 59 (19%) episodes diagnosed as VT. The causes of inappropriate detection were (1) algorithm failure (inability to fulfill the PR/=16 microg/ml) have been predominantly characterized with a C-->T transition in codon 86 of gyrA. The gyrA gene encodes one subunit of DNA gyrase, which is a primary target for fluoroquinolone antibiotics. This study establishes a rapid PCR-based TaqMan method for identifying ciprofloxacin-resistant C. jejuni strains that carry the C ->T transition in codon 86 of gyrA. The assay uses real-time detection, eliminating the need for gel electrophoresis. Optimization of the assay parameters using purified Campylobacter DNA resulted in the ability to detect femtogram levels of DNA. The method should be useful for monitoring the development of ciprofloxacin resistance in C. jejuni. Compiled nucleotide sequence data on the quinolone resistance-determining region of gyrA in Campylobacter indicate that sequence comparison of this region is a useful method for tentative identification of Campylobacter isolates at the species level. PMID- 11060055 TI - Improved immunodiagnosis of cystic hydatid disease by using a synthetic peptide with higher diagnostic value than that of its parent protein, Echinococcus granulosus antigen B. AB - The assays are used for the diagnosis of hydatid disease are still imperfect. The reported diagnostic sensitivity and specificity vary greatly depending on the panel of sera used, the laboratory conducting the assay, and, more critically, the antigen used. To contribute to its standardization, we have recently ranked the diagnostic performances of the major parasite antigens and the available synthetic peptides using a large collection of serum samples. That work showed that antigen B (AgB) possesses the highest diagnostic value among these antigens. In the present work we further dissected its antigenicity by analyzing the reactivity of the same panel of sera against a set of synthetic peptides spanning the sequence of both AgB subunits. The N-terminal extension of these subunits appeared to be immunodominant in human infections. A 38-mer peptide (p176) delineated from the N-terminal extension of the AgB/1 subunit performed in an enzyme-linked immunosorbent assay with a higher diagnostic sensitivity (80%) and specificity (94%) than native AgB, Ag5, or any other peptide antigen tested against this collection of serum samples. In view of its high diagnostic value and its nature as a well-defined reproducible antigen, p176 could conveniently be used as a reference standard antigen in the diagnosis of hydatid disease. PMID- 11060056 TI - Molecular epidemiological analysis of Cryptosporidium spp. in the United Kingdom: results of genotyping Cryptosporidium spp. in 1,705 fecal samples from humans and 105 fecal samples from livestock animals. AB - Cryptosporidium present in 1,705 fecal samples from humans and 105 from livestock animals were analyzed by PCR-restriction fragment length polymorphism of the Cryptosporidium oocyst wall protein. Overall, genotype 1 (human exclusive type) was detected in 37.8% of the samples from humans, genotype 2 (broad host range) was detected in 61.5%, a third genotype designated genotype 3 (Cryptosporidium meleagridis) was detected in 0.3%, and both genotypes 1 and 2 were recovered from 0.4%. All samples from livestock yielded genotype 2. Among 469 patients infected during eight drinking water-related outbreaks, five outbreaks were predominantly due to genotype 1, and three were due to genotype 2. Fifty-four samples were collected from patients involved with five swimming pool-associated outbreaks: two outbreaks were due to genotype 1, one was due to genotype 2, and the remaining two involved both genotypes 1 and 2. Among 26 family outbreaks and 1 children's nursery outbreak (2 to 3 members per group), the same genotype was recovered from the different members of each outbreak: 13 were due to genotype 1, and 14 were due to genotype 2. In eighteen patients reporting contact with animals and/or farms, genotype 1 was recovered from one patient and genotype 2 was recovered from the remaining 17. Among the sporadic cases, there were distinct geographical and temporal variations in the distribution of the genotypes. The spring peak in cases was due to genotype 2. Genotype 1 was significantly more common in patients infected during the late-summer-autumn peak and in those with a history of foreign travel. PMID- 11060057 TI - Diversity of domain V of 23S rRNA gene sequence in different Enterococcus species. AB - The highly conserved central loop of domain V of 23S RNA (nucleotides 2042 to 2628; Escherichia coli numbering) is implicated in peptidyltransferase activity and represents one of the target sites for macrolide, lincosamide, and streptogramin B antibiotics. DNA encoding domain V (590 bp) of several species of Enterococcus was amplified by PCR. Twenty enterococcal isolates were tested, including Enterococcus faecium (six isolates), Enterococcus faecalis, Enterococcus avium, Enterococcus durans, Enterococcus gallinarum, Enterococcus casseliflavus (two isolates of each), and Enterococcus raffinosus, Enterococcus mundtii, Enterococcus malodoratus, and Enterococcus hirae (one isolate of each). For all isolates, species identification by biochemical testing was corroborated by 16S rRNA gene sequencing. The sequence of domain V of the 23S rRNA gene from E. faecium and E. faecalis differed from those of all other enterococci. The domain V sequences of E. durans and E. hirae were identical. This was also true for E. gallinarum and E. casseliflavus. E. avium differed from E. casseliflavus by 23 bases, from E. durans by 16 bases, and from E. malodoratus by 2 bases. E. avium differed from E. raffinosus by one base. Despite the fact that domain V is considered to be highly conserved, substantial differences were identified between several enterococcal species. PMID- 11060058 TI - Analytical performance and clinical utility of a nucleic acid sequence-based amplification assay for detection of cytomegalovirus infection. AB - A nucleic acid sequence-based amplification (NASBA) assay for qualitative detection of human cytomegalovirus (CMV) pp67 mRNA was evaluated in a multicenter study. Negative results were obtained for all specimens from 50 CMV-seronegative and 50 CMV-seropositive low-risk whole-blood donors. No interference with CMV mRNA amplification was observed in the testing of 288 specimens containing various potential interfering substances, nonspecifically reacting substances (including mRNA from other herpesviruses), and three anticoagulants. A total of 95% (50 of 51) of CMV-positive (cell culture- and antigenemia immunofluorescence [AG-IFA]-positive) clinical specimens were positive by the NASBA assay. Results from different operators over multiple testing days were consistent for each of four panel members containing different concentrations of CMV mRNA, indicating the reproducibility of the assay. The estimated 95% reliable upper detection limit of the assay was 600 mRNA copies; the lower limit of detection was less than 25 mRNA copies. The clinical utility of the assay was evaluated with longitudinally collected specimens from solid-organ transplant patients (n = 21). A total of 98% (81 of 83) of the specimens from CMV-negative patients were negative by the NASBA assay, while 90% (10 of 11) of patient specimens that were positive by cell culture or AG-IFA were positive by the NASBA assay. Positive NASBA assay results were obtained earlier than AG-IFA or cell culture results for 55% of the patients and at the same time for the remainder of the patients (45%). The overall agreement between the NASBA assay and current reference tests was 86% when active CMV infection was present. These studies indicate that the CMV pp67 mRNA NASBA assay has reproducible and sensitive performance characteristics that should enable more rapid diagnosis of CMV infection. PMID- 11060059 TI - Evaluation of an immunocapture-agglutination test (Brucellacapt) for serodiagnosis of human brucellosis. AB - We evaluated the validity and the usefulness of a new test for the diagnosis of human brucellosis based on an immunocapture-agglutination technique. A total of 315 sera from 82 patients with a diagnosis of brucellosis, 157 sera from patients in whom brucellosis was suspected but not confirmed, and 412 sera from people living in rural areas with endemic brucellosis were studied. The seroagglutination test (SAT), Coombs anti-Brucella test, and Brucellacapt test were evaluated. All the initial sera from the 82 patients proved to be positive in Brucellacapt and Coombs tests, while only 75 (91.4%) were positive in the SAT. If a >/=1/160 diagnostic threshold titer was defined for the Brucellacapt test, Coombs test, and SAT, the sensitivities were 95.1, 91.5, and 65.8%, respectively. Taking the same diagnostic threshold titer for the 157 sera from the unconfirmed but suspected patients, the specificities of the Brucellacapt, Coombs, and SAT were 81.5, 96.2, and 100%, respectively; for the 412 control sera, the specificities were 99.0, 99.8, and 100%. The diagnostic efficiency (area below the receiver operating characteristic curve) of Brucellacapt was 0.987852 (95% confidence interval [CI], 0.95109 to 0.99286), very similar to the diagnostic efficiency of the Coombs test (0.97611; 95% CI, 0.94781 to 0.99146) and higher than that of SAT (0.91013; 95% CI, 0.86649 to 0.94317). The results of the Brucellacapt test were compared with those of the Coombs test (correlation coefficient, 0.956; P = 0.000) and SAT (correlation coefficient, 0.866; P = 0.000). The study shows very good correlation between the Brucellacapt and Coombs tests, with a high concordance between titers obtained in the two tests. Nevertheless, lower correlation and concordance were found between the Brucellacapt and Coombs tests when the results for titers of >/=1/160 were compared (0.692; P = 0.000). In acute brucellosis, the Brucellacapt and Coombs tests render positive titers of >/=1/160. When the titers are lower, they increase significantly in the following 30 days, despite the evolution of SAT titers. In contrast, Brucellacapt and Coombs titers are always high (>/=1/640) in brucellosis with long evolution, whether SAT titers are higher or lower than 1/160. PMID- 11060060 TI - Detection of cytomegalovirus DNA in human specimens by LightCycler PCR. AB - Detection of human cytomegalovirus (CMV) DNA in clinical specimens is considered a cornerstone in the diagnosis of CMV disease. The aim of this study was to evaluate a newly designed LightCycler-based quantitative CMV PCR. Specimens of human origin (n = 200) were tested using the LightCycler PCR, the quantitative COBAS AMPLICOR CMV MONITOR (CACM) assay, and a qualitative in-house PCR assay for the presence of CMV DNA. Samples that were reactive in at least two of the three assays were considered CMV DNA positive (n = 95 [47. 5%]), while samples that were nonreactive in two of the three assays were considered CMV DNA negative (n = 105 [52.5%]). Using the LightCycler assay, CMV DNA was detected in 91 of the 95 CMV DNA-positive human specimens (sensitivity, 95.8%; 95% confidence interval [CI], 89.6 to 98.8) and in 1 of the CMV DNA-negative specimens (specificity, 99%; 95% CI, 94.8 to 99.8). Results of CMV load determination as assessed by both quantitative test systems were correlated (r = 0.73; P < 0.0001; 95% CI, 0.61 to 0.81). Results for undiluted samples containing a high CMV load were more accurate with the LightCycler test than were results obtained with the CACM test, which underestimated the viral load of samples containing high DNA copy numbers. The high level of sensitivity, specificity, accuracy, and rapidity provided by the LightCycler technology are favorable for the use of this system in the detection of CMV DNA in clinical specimens. PMID- 11060061 TI - Development of a highly sensitive and specific enzyme-linked immunosorbent assay based on recombinant matrix protein for detection of avian pneumovirus antibodies. AB - The matrix (M) protein of avian pneumovirus (APV) was evaluated for its antigenicity and reliability in an enzyme-linked immunosorbent assay (ELISA) for diagnosis of APV infection, a newly emergent disease of turkeys in United States. Sera from APV-infected turkeys consistently contained antibodies to a 30-kDa protein (M protein). An ELISA based on recombinant M protein generated in Escherichia coli was compared with the routine APV ELISA that utilizes inactivated virus as antigen. Of 34 experimentally infected turkeys, 33 (97.1%) were positive by M protein ELISA whereas only 18 (52.9%) were positive by routine APV ELISA 28 days after infection. None of the serum samples from 41 uninfected experimental turkeys were positive by M protein ELISA. Of 184 field sera from turkey flocks suspected of having APV infection, 133 (72.3%) were positive by M protein ELISA whereas only 99 (53.8%) were positive by routine APV ELISA. Twelve serum samples, which were negative by M protein ELISA but positive by routine APV ELISA, were not reactive with either recombinant M protein or denatured purified APV proteins by Western analysis. This indicates that the samples had given false positive results by routine APV ELISA. The M protein ELISA was over six times more sensitive than virus isolation (11.5%) in detecting infections from samples obtained from birds showing clinical signs of APV infection. Taken together, these results show that ELISA based on recombinant M protein is a highly sensitive and specific test for detecting antibodies to APV. PMID- 11060062 TI - Quality of human immunodeficiency virus viral load testing in Australia. AB - This study determined the proficiencies of laboratories measuring human immunodeficiency virus type 1 (HIV-1) viral loads and the accuracies of two assays used for HIV-1 viral load measurement in Australia and investigated the variability of the new versions of these assays. Quality assessment program panels containing (i) dilutions of HIV-1 subtype B, (ii) replicates of identical samples of HIV-1 subtype B, and (iii) samples of subtype E and B were tested by laboratories. Total variability (within and between laboratories) was tested with quality control samples. The coefficients of variation (CVs) for the Roche AMPLICOR HIV-1 MONITOR version (v) 1.0 and Chiron Quantiplex bDNA 2.0 assays ranged from 53 to 87% and 22 to 31%, respectively. The widespread occurrence of invalid runs with the AMPLICOR HIV-1 MONITOR 1.0 assay was identified. The CVs of the new versions of the assays were 82 to 86% for the AMPLICOR HIV-1 MONITOR v 1.5 assay and 16 to 23% for the Quantiplex bDNA 3.0 assay. For virus dilution samples, all but 5 of 19 laboratories obtained results within 2 standard deviations of the mean. The Quantiplex bDNA 2.0 assay reported values lower than those reported by the AMPLICOR HIV-1 MONITOR version 1.0 assay for samples containing HIV-1 subtype B, whereas the reverse was true for subtype E. Identification and resolution of the problem of invalid runs markedly improved the quality of HIV-1 viral load testing. The variability observed between laboratories and between assays, even the most recent versions, dictates that monitoring of viral load in an individual should always be by the same laboratory and by the same assay. Results for an individual which differ by less than 0.5 log(10) HIV-1 RNA copy number/ml should not be considered clinically significant. PMID- 11060063 TI - Serological relationships of Cryptococcus spp.: distribution of antigenic factors in Cryptococcus and intraspecies diversity. AB - The antigenic formulas of 34 species in the genus Cryptococcus were determined by using type strains and eight factor sera prepared from adsorption experiments with Cryptococcus neoformans serotypes. These antigenic factors were shared by 19 species. The strains used could be divided into eight serological groups. The patterns of groups 1, 2, 3, 5, and 6 were the same as the patterns of C. neoformans serotypes A, D, A-D, B, and C, respectively. The species belonging to group 4 reacted to factor sera 1, 2, and 3. Group 7 contained one species that reacted only to factor serum 1. The 15 species in group 8 did not react to any of the factor sera used. Compared to the reported molecular phylogenetic tree, the serological and phylogenetic data were correlated in the Filobasidium lineage. All the members of the albidus clade in the Filobasidium lineage had antigens 1, 2, and 3, and all the strains in the magnus clade belonged to serogroup 8. Moreover, intraspecies diversity was examined using strains of C. curvatus, C. humicolus, and C. laurentii. Serological heterogeneity was observed in the species C. humicolus and C. laurentii, as well as in phylogenetic relationships previously published. Using serological features, similarities and differences between Cryptococcus species were demonstrated. Our study contributes to a better description of the genus Cryptococcus and related species phenotypically and phylogenetically. PMID- 11060064 TI - Molecular and pathogenic characterization of Borrelia burgdorferi sensu lato isolates from Spain. AB - Fifteen Borrelia burgdorferi sensu lato isolates from questing ticks and skin biopsy specimens from erythema migrans patients in three different areas of Spain were characterized. Four different genospecies were found (nine Borrelia garinii, including the two human isolates, three B. burgdorferi sensu stricto, two B. valaisiana, and one B. lusitaniae), showing a diverse spectrum of B. burgdorferi sensu lato species. B. garinii isolates were highly variable in terms of pulsed field gel electrophoresis pattern and OspA serotype, with four of the seven serotypes described. One of the human isolates was OspA serotype 5, the same found in four of seven tick isolates. The second human isolate was OspA serotype 3, which was not present in ticks from the same area. Seven B. garinii isolates were able to disseminate through the skin of C3H/HeN mice and to cause severe inflammation of joints. One of the two B. valaisiana isolates also caused disease in mice. Only one B. burgdorferi sensu stricto isolate was recovered from the urinary bladder. One isolate each of B. valaisiana and B. lusitaniae were not able to disseminate through the skin of mice or to infect internal organs. In summary, there is substantial diversity in the species and in the pathogenicity of B. burgdorferi sensu lato in areas in northern Spain where Lyme disease is endemic. PMID- 11060065 TI - Multicenter comparison of Roche COBAS AMPLICOR MONITOR version 1.5, Organon Teknika NucliSens QT with Extractor, and Bayer Quantiplex version 3.0 for quantification of human immunodeficiency virus type 1 RNA in plasma. AB - The performance and characteristics of Roche COBAS AMPLICOR HIV-1 MONITOR version 1.5 (CA MONITOR 1.5) UltraSensitive (usCA MONITOR 1. 5) and Standard (stCA MONITOR 1.5) procedures, Organon Teknika NucliSens HIV-1 RNA QT with Extractor (NucliSens), and Bayer Quantiplex HIV RNA version 3.0 (bDNA 3.0) were compared in a multicenter trial. Samples used in this study included 460 plasma specimens from human immunodeficiency virus (HIV) type 1 (HIV-1)-infected persons, 100 plasma specimens from HIV antibody (anti-HIV)-negative persons, and culture supernatants of HIV-1 subtype A to E isolates diluted in anti-HIV-negative plasma. Overall, bDNA 3.0 showed the least variation in RNA measures upon repeat testing. For the Roche assays, usCA MONITOR 1.5 displayed less variation in RNA measures than stCA MONITOR 1.5. NucliSens, at an input volume of 2 ml, showed the best sensitivity. Deming regression analysis indicated that the results of all three assays were significantly correlated (P < 0.0001). However, the mean difference in values between CA MONITOR 1.5 and bDNA 3.0 (0.274 log(10) RNA copies/ml; 95% confidence interval, 0.192 to 0.356) was significantly different from 0, indicating that CA MONITOR 1.5 values were regularly higher than bDNA 3.0 values. Upon testing of 100 anti-HIV-negative plasma specimens, usCA MONITOR 1.5 and NucliSens displayed 100% specificity, while bDNA 3.0 showed 98% specificity. NucliSens quantified 2 of 10 non-subtype B viral isolates at 1 log(10) lower than both CA MONITOR 1.5 and bDNA 3.0. For NucliSens, testing of specimens with greater than 1,000 RNA copies/ml at input volumes of 0.1, 0.2, and 2.0 ml did not affect the quality of results. Additional factors differing between assays included specimen throughput and volume requirements, limit of detection, ease of execution, instrument work space, and costs of disposal. These characteristics, along with assay performance, should be considered when one is selecting a viral load assay. PMID- 11060066 TI - Real-time quantitative PCR for human herpesvirus 6 DNA. AB - The diagnosis of human herpesvirus 6 (HHV-6) infection represents a complex issue because the most widely used diagnostic tools, such as immunoglobulin G antibody titer determination and qualitative DNA PCR with blood cells, are unable to distinguish between latent (clinically silent) and active (often clinically relevant) infection. We have developed a new, highly sensitive, quantitative PCR assay for the accurate measurement of HHV-6 DNA in tissue-derived cell suspensions and body fluids. The test uses a 5' nuclease, fluorogenic assay combined with real-time detection of PCR amplification products with the ABI PRISM 7700 sequence detector system. The sensitivity of this method is equal to the sensitivity of a nested PCR protocol (lower detection limit, 1 viral genome equivalent/test) for both the A and the B HHV-6 subgroups and shows a wider dynamic range of detection (from 1 to 10(6) viral genome equivalents/test) and a higher degree of accuracy, repeatability, and reproducibility compared to those of a standard quantitative-competitive PCR assay developed with the same reference DNA molecule. The novel technique is versatile, showing the same sensitivity and dynamic range with viral DNA extracted from different fluids (i.e., culture medium or plasma) or from tissue-derived cell suspensions. Furthermore, by virtue of its high-throughput format, this method is well suited for large epidemiological surveys. PMID- 11060067 TI - Human T-cell lymphotropic virus type 1 gag indeterminate western blot patterns in Central Africa: relationship to Plasmodium falciparum infection. AB - To gain insight on the significance of human T-cell lymphotropic virus type 1 (HTLV-1) indeterminate serological reactivities, we studied villagers of South Cameroon, focusing on a frequent and specific HTLV-1 Gag indeterminate profile (HGIP) pattern (gag p19, p26, p28, and p30 without p24 or Env gp21 and gp46). Among the 102 sera studied, 29 from all age groups had a stable HGIP pattern over a period of 4 years. There was no epidemiological evidence for sexual or vertical transmission of HGIP. Seventy-five percent of HGIP sera reacted positively on MT2 HTLV-1-infected cells by immunofluorescence assay. However, we could not isolate any HTLV-1 virus or detect the presence of p19 Gag protein in cultures of peripheral blood mononuclear cells obtained from individuals with strong HGIP reactivity. PCR experiments conducted with primers for HTLV-1 and HTLV-2 (HTLV 1/2 primers) encompassing different regions of the virus did not yield HTLV-1/2 proviral sequences from individuals with HGIP. Using 11 peptides corresponding to HTLV-1 or HTLV-2 immunodominant B epitopes in an enzyme-linked immunosorbent assay, one epitope corresponding to the Gag p19 carboxyl terminus was identified in 75% of HGIP sera, while it was recognized by only 41% of confirmed HTLV-1 positive sera. A positive correlation between HTLV-1 optical density values and titers of antibody to Plasmodium falciparum was also demonstrated. Finally, passage of sera through a P. falciparum-infected erythrocyte-coupled column was shown to specifically abrogate HGIP reactivity but not the HTLV-1 pattern, suggesting the existence of cross-reactivity between HTLV-1 Gag proteins and malaria-derived antigens. These data suggest that in Central Africa, this frequent and specific Western blot is not caused by HTLV-1 infection but could instead be associated with P. falciparum infection. PMID- 11060068 TI - Evaluation of fluorescence-based amplified fragment length polymorphism analysis for molecular typing in hospital epidemiology: comparison with pulsed-field gel electrophoresis for typing strains of vancomycin-resistant Enterococcus faecium. AB - Fluorescence-based amplified fragment length polymorphism (fbAFLP) is a novel assay based on the fluorescent analysis of an amplified subset of restriction fragments. The fbAFLP assay involves the selective PCR amplification of restriction fragments from a total digest of genomic DNA. The ligation of adapters with primer-specific sites coupled with primers containing selective nucleotides allowed the full potential of PCR to be realized while maintaining the advantages of restriction endonuclease analysis. Fluorescence-based fragment analysis with polyacrylamide gel electrophoresis provides the accurate band sizing required for homology assessment. The large number of phylogenetically informative characters obtained by fbAFLP is well suited for cluster analysis and database development. The method demonstrated excellent reproducibility and ease of performance and interpretation. We typed 30 epidemiologically well characterized isolates of vancomycin-resistant enterococci from an outbreak in a university hospital by fbAFLP. Clustering of fbAFLP data matched epidemiological, microbiological, and pulsed-field gel electrophoresis data. This study demonstrates the unprecedented utility of fbAFLP for epidemiological investigation. Future developments in standardization and automation will set fbAFLP as the "gold standard" for molecular typing in epidemiology. PMID- 11060069 TI - Rapid detection of west nile virus from human clinical specimens, field-collected mosquitoes, and avian samples by a TaqMan reverse transcriptase-PCR assay. AB - The authors report on the development and application of a rapid TaqMan assay for the detection of West Nile (WN) virus in a variety of human clinical specimens and field-collected specimens. Oligonucleotide primers and FAM- and TAMRA-labeled WN virus-specific probes were designed by using the nucleotide sequence of the New York 1999 WN virus isolate. The TaqMan assay was compared to a traditional reverse transcriptase (RT)-PCR assay and to virus isolation in Vero cells with a large number ( approximately 500) of specimens obtained from humans (serum, cerebrospinal fluid, and brain tissue), field-collected mosquitoes, and avian tissue samples. The TaqMan assay was specific for WN virus and demonstrated a greater sensitivity than the traditional RT-PCR method and correctly identified WN virus in 100% of the culture-positive mosquito pools and 98% of the culture positive avian tissue samples. The assay should be of utility in the diagnostic laboratory to complement existing human diagnostic testing and as a tool to conduct WN virus surveillance in the United States. PMID- 11060070 TI - Detection and quantification of Plasmodium falciparum in blood samples using quantitative nucleic acid sequence-based amplification. AB - A quantitative nucleic acid sequence-based amplification (QT-NASBA) assay for the detection of Plasmodium parasites has been developed. Primers and probes were selected on the basis of the sequence of the small-subunit rRNA gene. Quantification was achieved by coamplification of the RNA in the sample with one modified in vitro RNA as a competitor in a single-tube NASBA reaction. Parasite densities ranging from 10 to 10(8) Plasmodium falciparum parasites per ml could be demonstrated and quantified in whole blood. This is approximately 1,000 times more sensitive than conventional microscopy analysis of thick blood smears. Comparison of the parasite densities obtained by microscopy and QT-NASBA with 120 blood samples from Kenyan patients with clinical malaria revealed that for 112 of 120 (93%) of the samples results were within a 1-log difference. QT-NASBA may be especially useful for the detection of low parasite levels in patients with early stage malaria and for the monitoring of the efficacy of drug treatment. PMID- 11060071 TI - Diagnostic performance of the Roche AMPLICOR PCR in detecting Neisseria gonorrhoeae in genitourinary specimens from female sex workers in Cotonou, Benin. AB - The objective of this study was to evaluate the diagnostic performance of the Roche multiplex AMPLICOR Chlamydia trachomatis/Neisseria gonorrhoeae PCR test for the detection of Neisseria gonorrhoeae infection in female urine specimens and wet and dry endocervical swabs. Endocervical swabs and urine specimens were collected from 342 female sex workers from Cotonou, Benin, and were tested using the AMPLICOR C. trachomatis/N. gonorrhoeae test (Roche Diagnostic Systems, Inc., Branchburg, N.J.) with internal control detection. Endocervical swabs were also cultured on Thayer-Martin medium. A series of alternate standards that included a combination of all the tests but not the test being evaluated was used to assess the performance of the test with each type of specimen. The sensitivity, specificity, and positive and negative predictive values for the urine were 53.8, 98.9, 93.5, and 87.5%, respectively. Corresponding figures for the wet swab were 91.5, 100, 100, and 97.4%, respectively. Those for the dry swab were 96.3, 96.2, 88.5, and 98.8%, respectively. Based on this study, the AMPLICOR PCR assay showed a low sensitivity for detection of N. gonorrhoeae infection in urine specimens, whereas the test was found to be highly sensitive and specific with endocervical specimens. PMID- 11060072 TI - Detection and identification of mycobacteria by amplification of the internal transcribed spacer regions with genus- and species-specific PCR primers. AB - We evaluated the usefulness of PCR assays that target the internal transcribed spacer (ITS) region for identifying mycobacteria at the species level. The conservative and species-specific ITS sequences of 33 species of mycobacteria were analyzed in a multialignment analysis. One pair of panmycobacterial primers and seven pairs of mycobacterial species-specific primers were designed. All PCRs were performed under the same conditions. The specificities of the primers were tested with type strains of 20 mycobacterial species from the American Type Culture Collection; 205 clinical isolates of mycobacteria, including 118 Mycobacterium tuberculosis isolates and 87 isolates of nontuberculous mycobacteria from 10 species; and 76 clinical isolates of 28 nonmycobacterial pathogenic bacterial species. PCR with the panmycobacterial primers amplified fragments of approximately 270 to 400 bp in all mycobacteria. PCR with the M. tuberculosis complex-specific primers amplified an approximately 120-bp fragment only for the M. tuberculosis complex. Multiplex PCR with the panmycobacterial primers and the M. tuberculosis complex-specific primers amplified two fragments that were specific for all mycobacteria and the M. tuberculosis complex, respectively. PCR with M. avium complex-, M. fortuitum-, M. chelonae-, M. gordonae-, M. scrofulaceum-, and M. szulgai-specific primers amplified specific fragments only for the respective target organisms. These novel primers can be used to detect and identify mycobacteria simultaneously under the same PCR conditions. Furthermore, this protocol facilitates early and accurate diagnosis of mycobacteriosis. PMID- 11060073 TI - Emergence and rapid spread of carbapenem resistance during a large and sustained hospital outbreak of multiresistant Acinetobacter baumannii. AB - Beginning in 1992, a sustained outbreak of multiresistant Acinetobacter baumannii infections was noted in our 1,000-bed hospital in Barcelona, Spain, resulting in considerable overuse of imipenem, to which the organisms were uniformly susceptible. In January 1997, carbapenem-resistant (CR) A. baumannii strains emerged and rapidly disseminated in the intensive care units (ICUs), prompting us to conduct a prospective investigation. It was an 18-month longitudinal intervention study aimed at the identification of the clinical and microbiological epidemiology of the outbreak and its response to a multicomponent infection control strategy. From January 1997 to June 1998, clinical samples from 153 (8%) of 1,836 consecutive ICU patients were found to contain CR A. baumannii. Isolates were verified to be A. baumannii by restriction analysis of the 16S-23S ribosomal genes and the intergenic spacer region. Molecular typing by repetitive extragenic palindromic sequence-based PCR and pulsed-field gel electrophoresis showed that the emergence of carbapenem resistance was not by the selection of resistant mutants but was by the introduction of two new epidemic clones that were different from those responsible for the endemic. Multivariate regression analysis selected those patients with previous carriage of CR A. baumannii (relative risk [RR], 35.3; 95% confidence interval [CI], 7.2 to 173.1), those patients who had previously received therapy with carbapenems (RR, 4.6; 95% CI, 1.3 to 15.6), or those who were admitted into a ward with a high density of patients infected with CR A. baumannii (RR, 1.7; 95% CI, 1.2 to 2.5) to be at a significantly greater risk for the development of clinical colonization or infection with CR A. baumannii strains. In accordance, a combined infection control strategy was designed and implemented, including the sequential closure of all ICUs for decontamination, strict compliance with cross-transmission prevention protocols, and a program that restricted the use of carbapenem. Subsequently, a sharp reduction in the incidence rates of infection or colonization with A. baumannii, whether resistant or susceptible to carbapenems, was shown, although an alarming dominance of the carbapenem-resistant clones was shown at the end of the study. PMID- 11060074 TI - Detection of influenza A viruses from different species by PCR amplification of conserved sequences in the matrix gene. AB - The recently raised awareness of the threat of a new influenza pandemic has stimulated interest in the detection of influenza A viruses in human as well as animal secretions. Virus isolation alone is unsatisfactory for this purpose because of its inherent limited sensitivity and the lack of host cells that are universally permissive to all influenza A viruses. Previously described PCR methods are more sensitive but are targeted predominantly at virus strains currently circulating in humans, since the sequences of the primer sets display considerable numbers of mismatches to the sequences of animal influenza A viruses. Therefore, a new set of primers, based on highly conserved regions of the matrix gene, was designed for single-tube reverse transcription-PCR for the detection of influenza A viruses from multiple species. This PCR proved to be fully reactive with a panel of 25 genetically diverse virus isolates that were obtained from birds, humans, pigs, horses, and seals and that included all known subtypes of influenza A virus. It was not reactive with the 11 other RNA viruses tested. Comparative tests with throat swab samples from humans and fecal and cloacal swab samples from birds confirmed that the new PCR is faster and up to 100-fold more sensitive than classical virus isolation procedures. PMID- 11060075 TI - Mycobacterium heckeshornense sp. nov., A new pathogenic slowly growing Mycobacterium sp. Causing cavitary lung disease in an immunocompetent patient. AB - A pathogenic scotochromogenic Mycobacterium xenopi-like organism was isolated from the lung of an immunocompetent young woman. This pathogen caused severe bilateral cavitary lung disease, making two surgical interventions necessary after years of chronic disease. This case prompted us to characterize this mycobacterium by a polyphasic taxonomic approach. The isolate contained chemotaxonomic markers which were typical for the genus Mycobacterium, i.e., the meso isomer of 2,6-diaminopimelic acid, arabinose, and galactose as diagnostic whole-cell sugars, MK-9(H(2)) as the principal isoprenoid quinone, a mycolic acid pattern of alpha-mycolates, ketomycolates, and wax ester mycolates, unbranched saturated and unsaturated fatty acids plus a significant amount of tuberculostearic acid, and small amounts of a C(20:0) secondary alcohol. On the basis of its unique 16S rRNA and 16S-23S spacer gene sequences, we propose that the isolate should be assigned to a new species, Mycobacterium heckeshornense. This novel species is phylogenetically closely related to M. xenopi. The type strain of M. heckeshornense is strain S369 (DSM 44428(T)). The GenBank accession number of the 16S rRNA gene of M. heckeshornense is AF174290. PMID- 11060076 TI - PCR detection of Escherichia coli O157:H7 directly from stools: evaluation of commercial extraction methods for purifying fecal DNA. AB - Rapid identification of Escherichia coli O157:H7 is important for patient management and for prompt epidemiological investigations. We evaluated one in house method and three commercially available kits for their ability to extract E. coli O157:H7 DNA directly from stool specimens for PCR. Of the 153 stool specimens tested, 107 were culture positive and 46 were culture negative. The sensitivities and specificities of the in-house enrichment method, IsoQuick kit, NucliSens kit, and QIAamp kit were comparable, as follows: 83 and 98%, 85 and 100%, 74 and 98%, and 86 and 100%, respectively. False-negative PCR results may be due to the presence of either inherent inhibitors or small numbers of organisms. The presence of large amounts of bacteria relative to the amount of the E. coli O157:H7 target may result in the lower sensitivities of the assays. All commercial kits were rapid and easy to use, although DNA extracted with the QIAamp kit did not require further dilution of the DNA template prior to PCR. PMID- 11060077 TI - Species-specific identification of human adenoviruses by a multiplex PCR assay. AB - A multiplex PCR assay was developed by using primers to the fiber gene that could differentiate human adenovirus (Ad) species A through F in a single amplification reaction. The assay correctly identified the species of all 49 recognized Ad prototype strains as well as 180 geographically and temporally diverse Ad field isolates. Ad serotype 6 (Ad6) (species C), Ad16 (species B), Ad31 (species A), and Ad40 and Ad41 (species F) could also be distinguished by amplicon size within each respective species. In comparison, a previously described Ad species specific multiplex PCR assay that used primers to the Ad hexon gene gave equivocal results with several serotypes of species B, whereas our multiplex assay amplified all species B serotypes equally well. Our multiplex PCR assay will permit rapid, accurate, and cost-effective classification of Ad isolates. PMID- 11060078 TI - Real-time PCR for quantitative detection of Toxoplasma gondii. AB - The protozoan Toxoplasma gondii is one of the most common infectious pathogenic parasites and can cause severe medical complications in infants and immunocompromised individuals. We report here the development of a real-time PCR based assay for the detection of T. gondii. Oligonucleotide primers and a fluorescence-labeled TaqMan probe were designed to amplify the T. gondii B1 gene. After 40 PCR cycles, the cycle threshold values (C(T)) indicative of the quantity of the target gene were determined. Typically, a C(T) of 25.09 was obtained with DNA from 500 tachyzoites of the T. gondii RH strain. The intra-assay coefficients of variation (CV) were 0.4, 0.16, 0.24, and 0.79% for the four sets of quadruplicate assays, with a mean interassay CV of 0.4%. These values indicate the reproducibility of this assay. Upon optimization of assay conditions, we were able to obtain a standard curve with a linear range (correlation coefficient = 0.9988) across at least 6 logs of DNA concentration. Hence, we were able to quantitatively detect as little as 0.05 T. gondii tachyzoite in an assay. When tested with 30 paraffin-embedded fetal tissue sections, 10 sections (33%) showed a C(T) of <40 and were scored as positive for this test. These results were consistent with those obtained through our nested-PCR control experiments. We have developed a rapid, sensitive, and quantitative real-time PCR for detection of T. gondii. The advantages of this technique for the diagnosis of toxoplasmosis in a clinical laboratory are discussed. PMID- 11060079 TI - Analysis of immunoreactivity to a Streptococcus equi subsp. zooepidemicus M-like protein To confirm an outbreak of poststreptococcal glomerulonephritis, and sequences of M-like proteins from isolates obtained from different host species. AB - The etiologic agent of a large 1998 outbreak of poststreptococcal acute glomerulonephritis (PSGN) in Nova Serrana, Brazil, was found likely to be a specific strain of Streptococcus equi subsp. zooepidemicus from contaminated cheese (S. Balter et al., Lancet 355:1776-1780, 2000). In the present study, we used a serologic screen for a known surface-exposed virulence factor to confirm the epidemiologic findings. Using primers flanking a previously characterized M like protein gene (J. F. Timoney et al., Infect. Immun. 63:1440-1445, 1995), we amplified and sequenced the M-like protein (designated Szp5058) gene and found it to be identical among four independent acute-phase PSGN patient isolates. Convalescent-phase sera from 33 of 44 patients in the PSGN outbreak were found to contain antibodies highly reactive to a purified Szp5058 fusion protein, compared with 1 of 17 control sera (P < 0. 0001), suggesting that Szp5058 was expressed during infection and further implicating this strain as the cause of the PSGN outbreak. The predicted signal sequence and cell wall association motif of Szp5058 were highly conserved with the corresponding sequence from S. equi subsp. zooepidemicus SzpW60, while the predicted surface-exposed portions differed markedly between these two proteins. The 5' end of the szp5058 gene, including its variable region, was identical to the szp gene from another strain associated with a previous PSGN outbreak in England (M. Barham et al., Lancet i:945-948, 1983), and the corresponding szp sequence found from the Lancefield group C type strain isolated from a guinea pig. In addition, the hypervariable (HV) portion of szp5058 was identical to a previously published HV sequence from a horse isolate (J. A. Walker and J. F. Timoney, Am. J. Vet. Res. 59:1129-1133, 1998). Three other strains of S. equi subsp. zooepidemicus, including another strain previously associated with a PSGN outbreak, were each found to contain a distinct szp gene. Two of these szp genes had HV regions identical to szp regions from isolates recovered from different host species. PMID- 11060080 TI - Outbreak of Bacillus cereus infections in a neonatal intensive care unit traced to balloons used in manual ventilation. AB - In 1998, an outbreak of systemic infections caused by Bacillus cereus occurred in the Neonatal Intensive Care Unit of the University Hospital Vrije Universiteit, Amsterdam, The Netherlands. Three neonates developed sepsis with positive blood cultures. One neonate died, and the other two neonates recovered. An environmental survey, a prospective surveillance study of neonates, and a case control study were performed, in combination with molecular typing, in order to identify potential sources and transmission routes of infection. Genotypic fingerprinting by amplified-fragment length polymorphism (AFLP) showed that the three infections were caused by a single clonal type of B. cereus. The same strain was found in trachea aspirate specimens of 35 other neonates. The case control study showed mechanical ventilation with a Sensormedics ventilation machine to be a risk factor for colonization and/or infection (odds ratio, 9.8; 95% confidence interval, 1.1 to 88.2). Prospective surveillance showed that colonization with B. cereus occurred exclusively in the respiratory tract of mechanically ventilated neonates. The epidemic strain of B. cereus was found on the hands of nursing staff and in balloons used for manual ventilation. Sterilization of these balloons ended the outbreak. We conclude that B. cereus can cause outbreaks of severe opportunistic infection in neonates. Typing by AFLP proved very useful in the identification of the outbreak and in the analysis of strains recovered from the environment to trace the cause of the epidemic. PMID- 11060081 TI - Genetic diversity and clonal patterns among antibiotic-susceptible and -resistant Streptococcus pneumoniae colonizing children: day care centers as autonomous epidemiological units. AB - Characterization by antibiotype of the 1,096 Streptococcus pneumoniae recovered from 2,111 nasopharyngeal samples of children attending 16 day care centers (DCCs) in Lisbon, Portugal, and molecular typing of 413 drug-resistant pneumococci (DRPn) and 89 fully drug-susceptible pneumococci (DSPn) has allowed several conclusions. (i) There was an increase in the frequency of DRPn colonizing children in DCCs from 40% in 1996 to 45% in 1997 to 50% in 1998. (ii) Drug resistance spread by cross-transmission of DRPn clones. A few (8 out of 57) DRPn clones were repeatedly isolated from a large number of children in several DCCs and during each period of surveillance, suggesting the epidemic nature of these clones, which included lineages representing internationally spread S. pneumoniae clones. (iii) Dissemination of resistance determinants among pneumococci colonizing the nasopharynx occurred. Association of identical pulsed field gel electrophoresis patterns with diverse antibiotypes among pneumococci colonizing children suggests that the high prevalence of DRPn involves not only cross-transmission of resistant strains but also dispersal of resistance genes through recombinational mechanisms. (iv) DCCs are autonomous epidemiological units. Among the 413 DRPn, 57 different lineages were detected; these lineages were dispersed among the 16 DCCs to produce unique microbiological profiles for each of the DCCs. Higher genetic diversity and less sharing of clonal types were observed among the DSPn. PMID- 11060082 TI - Rapid method for species-specific identification of Vibrio cholerae using primers targeted to the gene of outer membrane protein OmpW. AB - The distribution of genes for an outer membrane protein (OmpW) and a regulatory protein (ToxR) in Vibrio cholerae and other organisms was studied using respective primers and probes. PCR amplification results showed that all (100%) of the 254 V. cholerae strains tested were positive for ompW and 229 ( approximately 98%) of 233 were positive for toxR. None of the 40 strains belonging to other Vibrio species produced amplicons with either ompW- or toxR specific primers, while 80 bacterial strains from other genera tested were also found to be negative by the assay. These studies were extended with representative number of strains using ompW- and toxR-specific probes in DNA dot blot assay. While the V. cholerae strains reacted with ompW probe, only one (V. mimicus) out of 60 other bacterial strains tested showed weak recognition. In contrast, several strains belonging to other Vibrio species (e.g., V. mimicus, V. splendidus, V. alginolyticus, V. fluvialis, V. proteolyticus, V. aestuarianus, V. salmonicida, V. furnissii, and V. parahaemolyticus) showed weak to strong reactivity to the toxR probe. Restriction fragment length polymorphism analysis and nucleotide sequence data revealed that the ompW sequence is highly conserved among V. cholerae strains belonging to different biotypes and/or serogroups. All of these results suggest that the ompW gene can be targeted for the species specific identification of V. cholerae strains. The scope of this study was further extended through the development of a one-step multiplex PCR assay for the simultaneous amplification of ompW and ctxA genes which should be of considerable value in the screening of both toxigenic and nontoxigenic V. cholerae strains of clinical as well as environmental origin. PMID- 11060083 TI - Rapid detection of contagious caprine pleuropneumonia using a Mycoplasma capricolum subsp. capripneumoniae capsular polysaccharide-specific antigen detection latex agglutination test. AB - Latex microspheres (diameter, 8 microm) were coated with anti-Mycoplasma capricolum subsp. capripneumoniae polyclonal immunoglobulin G (IgG) antiserum (anti-F38 biotype). The coated microspheres, when used in a latex agglutination test (LAT), detected M. capricolum subsp. capripneumoniae antigen in the serum of goats with contagious caprine pleuropneumoniae (CCPP). Beads also agglutinated strongly in the presence of purified M. capricolum subsp. capripneumoniae capsular polysaccharide (CPS). Preabsorption of CPS-specific antibodies prior to coating of the beads removed agglutinating activity in the presence of M. capricolum subsp. capripneumoniae, strongly suggesting that CPS is the likely soluble antigen recognized by the test. In addition, the specificity of the LAT exactly mirrored that of an M. capricolum subsp. capripneumoniae CPS-specific monoclonal antibody (WM25): of the 8 other mycoplasma species tested, agglutination was observed only with bovine serogroup 7. The LAT detected all 11 strains of M. capricolum subsp. capripneumoniae examined in this study, with a sensitivity level of 2 ng of CPS, or the equivalent of 1.7 x 10(4) CFU, in a reaction volume of 0.03 ml of serum. With field sera from goats with CCPP, the results of the LAT exhibited a 67% correlation with the results of the currently used complement fixation test (CFT), with the main discrepancy in diagnosis resulting from the increased sensitivity of the LAT compared to that of CFT. This antigen-detection LAT should prove particularly useful in identifying animals in the earliest stages of CCPP and combines sensitivity and low cost with ease of application in the field, without the need for any specialist training or equipment. PMID- 11060084 TI - Characterization of a Borrelia burgdorferi VlsE invariable region useful in canine Lyme disease serodiagnosis by enzyme-linked immunosorbent assay. AB - Sera collected from dogs experimentally infected with Borrelia burgdorferi by tick inoculation were analyzed for an antibody response to each of the six invariable regions (IRs; i.e., IR(1) to IR(6)) of VlsE, the variable surface antigen of B. burgdorferi. Six synthetic peptides (C(1) to C(6)), which reproduced the six IR sequences were used as peptide-based, enzyme-linked immunosorbent assay (ELISA) antigens. Two IRs, IR(2) and IR(6), were found to be immunodominant. Studies with serially collected serum samples from experimentally infected dogs revealed that the antibody response to IR(6) appears earlier and is stronger than that to IR(2). Thus, the IR(6) sequence alone appeared to be sufficient for serodiagnosis. When C(6) alone was used as antigen, the peptide based ELISA was positive in 7 of 23 dogs (30%) as early as 3 weeks postinfection. All dogs (n = 33) became strongly positive 1 or 2 weeks later, and this response persisted for the entire study, which lasted for 69 weeks. Of 55 sera submitted by veterinarians from dogs suspected of having Lyme disease, 19 were also positive by the C(6) ELISA, compared to 20 positives detected by immunoblot analysis using cultured B. burgdorferi lysates as antigen. The sensitivity of using C(2) and C(6) together for detecting specific antibody in both experimentally infected and clinically diagnosed dogs was not better than sensitivity with C(6) alone, confirming that C(6) suffices as a diagnostic probe. Moreover, the C(6) ELISA yielded 100% specificity with serum samples collected from 70 healthy dogs, 14 dogs with infections other than B. burgdorferi, and 15 animals vaccinated with either outer surface protein A, whole-spirochete vaccines, or the common puppy-vaccines. Therefore, this C(6) ELISA was both sensitive and specific for the serodiagnosis of canine Lyme disease and could be used with vaccinated dogs. PMID- 11060085 TI - Comparison of the BACTEC MGIT 960 and ESP culture system II for growth and detection of mycobacteria. AB - The performances of two continuously monitoring mycobacterial culture systems-ESP Culture System II (ESP II; Trek Diagnostics, Inc. , Westlake, Ohio) and BACTEC MGIT 960 (BD Biosciences, Sparks, Md. )-were compared. In addition to both liquid media, all specimens were plated onto Middlebrook 7H11/7H11 selective agar. A total of 3, 151 specimens of all types (56.3% were respiratory specimens) were cultured; 231 (7.3%) yielded mycobacteria. The most common species recovered were Mycobacterium avium complex (69 isolates) and Mycobacterium tuberculosis complex (MTBC; 65 isolates). The recovery rates for ESP II, BACTEC MGIT 960, and Middlebrook agar, respectively, were 71.2, 63.9, and 61.8% for all mycobacteria; 70.2, 72.6, and 66.3% for all mycobacteria except Mycobacterium gordonae; and 73.8, 84.6, and 87.7% for MTBC. For liquid plus solid medium combinations, recovery rates for all mycobacteria and for MTBC, respectively, were 84.1 and 92.3% for ESP II plus Middlebrook agar and 81.5 and 98.5% for BACTEC MGIT 960 plus Middlebrook agar. The differences in recovery of all mycobacteria by ESP II and by BACTEC MGIT 960 were not significant; for the individual species, the only significant difference was recovery of more isolates of M. gordonae by ESP II. For those isolates recovered in both automated systems, mean times to detection of all mycobacteria and MTBC, respectively, were 15.8 and 17.4 days for ESP II and 12.5 and 11.9 days for BACTEC MGIT 960 (P < 0.05). False-positive signals occurred with 23 (0.7%) BACTEC MGIT 960 cultures and 84 (2.7%) ESP II cultures (P < 0.01). Overall contamination rates were 17.1% for BACTEC MGIT 960, 18.9% for ESP II, and 11.0% for Middlebrook agar. In summary, the ESP II and BACTEC MGIT 960 systems performed comparably with regard to growth and detection of mycobacteria, and the contamination rates were similar. However, with ESP II, times to detection of all mycobacteria and of MTBC were significantly longer, the recovery rate of M. gordonae was significantly higher, and the number of false positive signals was greater than with BACTEC MGIT 960. PMID- 11060086 TI - Improved version 2.0 qualitative and quantitative AMPLICOR reverse transcription PCR tests for hepatitis C virus RNA: calibration to international units, enhanced genotype reactivity, and performance characteristics. AB - Version 2.0 qualitative and quantitative AMPLICOR reverse transcription-PCR tests for HCV were designed to improve on the performance of first version of the hepatitis C virus (HCV) tests. The new tests were calibrated in international units, the new commonly accepted standard unit of measurement for HCV RNA. The sensitivity of the qualitative tests was enhanced by modifying the specimen processing procedure to achieve a limit of detection 50 IU/ml. The limit of detection for the quantitative tests was 600 IU/ml. Modifications to the amplification reaction mixture and thermal cycling conditions enabled all genotypes to be amplified with similar efficiency. The quantitative tests exhibited a linear range extending from 500 to 500,000 IU/ml and excellent reproducibility, with coefficients of variation ranging from 18 to 39%, within the linear range. These data indicate that the version 2. 0 AMPLICOR HCV tests will improve diagnosis of HCV infection and will yield more-accurate titers for prognosis and for monitoring therapeutic efficacy, particularly at low viral loads. Furthermore, it will be possible to compare the performance characteristics and viral load measurements of AMPLICOR tests to those of other tests that adopt the international unit as the standard of measurement. PMID- 11060087 TI - Evaluation of PCR-based methods for discrimination of Francisella species and subspecies and development of a specific PCR that distinguishes the two major subspecies of Francisella tularensis. AB - Previous studies have demonstrated that the four subspecies of the human pathogen Francisella tularensis, despite showing marked variations in their virulence for mammals and originating from different regions in the Northern Hemisphere, display a very close phylogenetic relationship. This property has hampered the development of generally applicable typing methods. To overcome this problem, we evaluated the use of PCR for discrimination of the subspecies using various forms of long arbitrary primers or primers specific for repetitive extragenic palindromic sequences (REP) or enterobacterial repetitive intragenic consensus (ERIC) sequences. Patterns generated by use of REP, ERIC, or long arbitrary primers allowed differentiation at the species level and of the four subspecies of F. tularensis. With each of these three methods, similar or identical clustering of strains was found, and groups of strains of different geographical origins or differing in virulence showed distinct patterns. The discriminatory indices of the methods varied from 0.57 to 0.65; thus, the patterns were not sufficiently discriminatory to distinguish individual strains. The sequence of a fragment generated by amplification with an arbitrary primer was determined, and a region showing interstrain heterogeneity was identified. Specific primers were designed, and a PCR was developed that distinguished strains of F. tularensis subsp. holarctica from strains of other F. tularensis subspecies, including strains of the highly virulent F. tularensis subsp. tularensis. Notably, one European isolate showed the genetic pattern typical of the highly virulent F. tularensis subsp. tularensis, generally believed to exist only in North America. It is proposed that a combination of the specific PCR together with one method generating subspecies-specific patterns is suitable as a rapid and relatively simple strategy for discrimination of Francisella species and subspecies. PMID- 11060088 TI - Panfungal PCR and multiplex liquid hybridization for detection of fungi in tissue specimens. AB - A procedure based on panfungal PCR and multiplex liquid hybridization was developed for the detection of fungi in tissue specimens. The PCR amplified the fungal internal transcribed spacer (ITS) region (ITS1-5.8S rRNA-ITS2). After capture with specific probes, eight common fungal pathogens (Aspergillus flavus, Aspergillus fumigatus, Candida albicans, Candida krusei, Candida glabrata, Candida parapsilosis, Candida tropicalis, and Cryptococcus neoformans) were identified according to the size of the amplification product on an automated sequencer. The nonhybridized products were identified by sequencing. The performance of the procedure was examined with 12 deep-tissue specimens and 8 polypous tissue biopsies from the paranasal sinuses. A detection level of 0.1 to 1 pg of purified DNA (2 to 20 CFU) was achieved. Of the 20 specimens, PCR was positive for 19 (95%), of which 10 (53%) were hybridization positive. In comparison, 12 (60%) of the specimens were positive by direct microscopy, but only 7 (35%) of the specimens showed fungal growth. Sequencing of the nonhybridized amplification products identified an infecting agent in six specimens, and three specimens yielded only sequences of unknown fungal origin. The procedure provides a rapid (within 2 days) detection of common fungal pathogens in tissue specimens, and it is highly versatile for the identification of other fungal pathogens. PMID- 11060089 TI - Coyotes (Canis latrans) as the reservoir for a human pathogenic Bartonella sp.: molecular epidemiology of Bartonella vinsonii subsp. berkhoffii infection in coyotes from central coastal California. AB - Bartonella vinsonii subsp. berkhoffii was originally isolated from a dog suffering infectious endocarditis and was recently identified as a zoonotic agent causing human endocarditis. Following the coyote bite of a child who developed clinical signs compatible with Bartonella infection in Santa Clara County, Calif., this epidemiological study was conducted. Among 109 coyotes (Canis latrans) from central coastal California, 31 animals (28%) were found to be bacteremic with B. vinsonii subsp. berkhoffii and 83 animals (76%) had B. vinsonii subsp. berkhoffii antibodies. These findings suggest these animals could be the wildlife reservoir of B. vinsonii subsp. berkhoffii. PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of the gltA and 16S rRNA genes for these 31 isolates yielded similar profiles that were identical to those of B. vinsonii subsp. berkhoffii. Partial sequencing of the gltA and 16S rRNA genes, respectively, indicated 99.5 and 100% homology between the coyote isolate and B. vinsonii subsp. berkhoffii (ATCC 51672). PCR-RFLP analysis of the 16S-23S intergenic spacer region showed the existence of two different strain profiles, as has been reported in dogs. Six (19%) of 31 Bartonella bacteremic coyotes exhibited the strain profile that was identified in the type strain of a canine endocarditis case (B. vinsonii subsp. berkhoffii ATCC 51672). The other 25 bacteremic coyotes were infected with a strain that was similar to the strains isolated from healthy dogs. Based on whole bacterial genome analysis by pulsed field gel electrophoresis (PFGE) with SmaI restriction endonuclease, there was more diversity in fingerprints for the coyote isolates, which had at least 10 major variants compared to the two variants described for domestic dog isolates from the eastern United States. By PFGE analysis, three Bartonella bacteremic coyotes were infected by a strain identical to the one isolated from three healthy dog carriers. Further studies are necessary to elucidate the mode of transmission of B. vinsonii subsp. berkhoffii, especially to identify potential vectors, and to determine how humans become infected. PMID- 11060090 TI - Application of tRNA intergenic spacer PCR for identification of Enterococcus species. AB - tRNA intergenic spacer PCR (tDNA-PCR) was evaluated for its usefulness in the differentiation of enterococcal species of human and animal origin. This technique was carried out for 124 strains belonging to 17 enterococcal species and generated DNA fragments, which were separated by capillary electrophoresis. tDNA-PCR enabled us to discriminate for all species tested. Enterococcus faecium showed minor but reproducible differences with Enterococcus durans, while Enterococcus hirae was easily distinguishable. Enterococcus avium, Enterococcus malodoratus, and Enterococcus raffinosus generated highly similar though distinctive patterns. PMID- 11060091 TI - Granulocytic Ehrlichiae in Ixodes persulcatus ticks from an area in China where Lyme disease is endemic. AB - A total of 372 adult Ixodes persulcatus ticks were collected from vegetation in a forest area of Heilongjiang Province in northeastern China, where Lyme disease is known to be endemic. The ticks were examined for the presence of granulocytic ehrlichiae by heminested PCR with primers derived from the 16S rRNA gene. Of 310 ticks obtained from the Dahe forestry farm, two pools (each containing 5 ticks) were found positive, with a minimum infection rate of 0.6%. Ehrlichial DNA was also detected in one female (1.6%) of 62 ticks collected from the Yulin forestry farm. The overall minimum infection rate of the 372 I. persulcatus adults was 0.8%. The nucleotide sequences of 919-bp PCR products from the three positive tick specimens were identical to each other and very closely related to the members of the Ehrlichia phagocytophila genogroup. This is the first identification of granulocytic ehrlichiae in ticks in Asia and the first report of infection in I. persulcatus anywhere. PMID- 11060092 TI - Comparison of intertypic antigenicity of Aino virus isolates by dot immunobinding assay using neutralizing monoclonal antibodies. AB - Neutralizing monoclonal antibodies (MAbs) against the Aino virus were prepared, and the neutralizing epitopes of the virus were defined by competitive binding assay. Seven continuous and overlapping neutralizing epitopes existed on the G1 glycoprotein of the Aino virus. Two antigenic domains were identified and were designated I and II, with domain II consisting of six epitopes. Dot immunobinding assays (DIAs) were performed with MAbs that recognized these seven neutralizing epitopes. DIAs were performed with 1 Australian strain and 21 isolates found in Japan between the years 1964 and 1995. The MAb response patterns of all isolates were divided into four groups. The Japanese isolates did not show large differences in antigenicity, but the antigenicity of the Australian strain collected in 1968 was significantly different from that of the Japanese strains; the Australian strain lacked reactivity to three epitopes and showed only low reactivity to one epitope. PMID- 11060093 TI - Detection of Legionella pneumophila using a real-time PCR hybridization assay. AB - A real-time PCR hybridization assay for Legionella pneumophila is described; the assay uses LightCycler (Idaho Technology) methodology to specifically detect 2.5 CFU/reaction, equivalent to 1,000 CFU/liter of starting water sample. The assay, including DNA extraction and confirmation of product identity, is completed within 90 min of receipt of a sample. PMID- 11060094 TI - Detection of Ehrlichia platys DNA in brown dog ticks (Rhipicephalus sanguineus) in Okinawa Island, Japan. AB - Thirty-two Rhipicephalus sanguineus females collected from eight free-roaming dogs in Okinawa Island, Japan, were examined for ehrlichial DNA by 16S rRNA-based PCR and subsequent sequencing. Partial sequences of Ehrlichia platys 16S rRNA (678 to 679 bp) were detected in three ticks (9.4%) from two dogs. This is the first report of detection of E. platys in Japan, and also the first report of detection in ticks. PMID- 11060095 TI - Extensive sequence divergence among bovine respiratory syncytial viruses isolated during recurrent outbreaks in closed herds. AB - The nucleotides coding for the extracellular part of the G glycoprotein and the full SH protein of bovine respiratory syncytial virus (BRSV) were sequenced from viruses isolated from numerous outbreaks of BRSV infection. The isolates included viruses isolated from the same herd (closed dairy farms and veal calf production units) in different years and from all confirmed outbreaks in Denmark within a short period. The results showed that identical viruses were isolated within a herd during outbreaks and that viruses from recurrent infections varied by up to 11% in sequence even in closed herds. It is possible that a quasispecies variant swarm of BRSV persisted in some of the calves in each herd and that a new and different highly fit virus type (master and consensus sequence) became dominant and spread from a single animal in connection with each new outbreak. Based on the high level of diversity, however, the most likely explanation was that BRSV was (re)introduced into the herd prior to each new outbreak. These findings are highly relevant for the understanding of the transmission patterns of BRSV among calves and human respiratory syncytial virus among humans. PMID- 11060096 TI - Detection of extended-spectrum beta-lactamases in klebsiellae with the Oxoid combination disk method. AB - The Oxoid combination disk method for detecting extended-spectrum beta-lactamases (ESBLs) depends on comparing the inhibition zones of cefpodoxime (10-microg) and cefpodoxime-plus-clavulanate (10- plus 1-microg) disks. The presence of clavulanate enlarged the zones for all of 180 ESBL-producing klebsiellae by >/=5 mm, whereas zones for cefpodoxime-susceptible isolates and cefpodoxime-resistant isolates with AmpC and K1 beta-lactamases were enlarged by Cs+ > Na+ for I(HA) and Na+ > K+ >> Cs+ for I(DA). I(HA) was activated and deactivated instantaneously and showed no inactivation whereas I(DA) was activated, inactivated and deactivated within tens of milliseconds. These currents were inhibited by external calcium with an IC50 of 0.3 microM for I(DA) and an IC50 of 20 microM for I(HA). Cyclopiazonic acid (CPA) induced an outward, but not an inward current. SK&F 96365, a blocker of store-operated Ca2+ channels, suppressed I(DA) with a half maximal inhibitory concentration of 9 microM but was ineffective in inhibiting I(HA) at concentrations up to 100 microM. Gd3+ and La3+ strongly suppressed I(DA) at 1 and 10 microM, respectively and were less effective in blocking I(HA) (complete inhibition required a concentration of 100 microM for both). Carbachol at 10-100 microM evoked about a 3-fold increase in I(HA) amplitude and completely abolished I(DA). We conclude that I(HA) and I(DA) are Ca2+-blockable cationic currents with different ion selectivity profiles that are carried by different channels. I(DA) shows novel voltage-dependent properties for a cationic current. PMID- 11060130 TI - Inactivation and tachyphylaxis of heat-evoked inward currents in nociceptive primary sensory neurones of rats. AB - Membrane currents evoked by repeated noxious heat stimuli (43-47 degrees C) of 3 s duration were investigated in acutely dissociated dorsal root ganglion (DRG) neurones of adult rats. The heat stimuli generated by a fast solution exchanger had a rise time of 114 +/- 6 ms and a fall time of 146 +/- 13 ms. When heat stimuli were applied to heat-sensitive small (< or = 32.5 microm) DRG neurones, an inward membrane current (I(heat)) with a mean peak of 2430 +/- 550 pA was observed (n = 19). This current started to activate and deactivate with no significant latency with respect to the heat stimulus. The peak of I(heat) was reached with a rise time of 625 +/- 115 ms. When the heat stimulus was switched off I(heat) deactivated with a fall time of 263 +/- 17 ms. During constant heat stimulation I(heat) decreased with time constants of 4-5 s (inactivation). At the end of a 3 s heat stimulus the peak current was reduced by 44 +/- 5 % (n = 19). Current-voltage curves revealed outward rectifying properties of I(heat) and a reversal potential of -6.3 +/- 2.2 mV (n = 6). Inactivation was observed at all membrane potentials investigated (-80 to 60 mV); however, inactivation was more pronounced for inward currents (37 +/- 5 %) than for outward currents (23 +/- 6 %, P < 0.05). When neurones were investigated with repeated heat stimuli (3 to 5 times) of the same temperature, the peak current relative to the first I(heat) declined by 48 +/- 6 % at the 3rd stimulus (n = 19) and by 54 +/- 18 % at the 5th stimulus (n = 4; tachyphylaxis). In the absence of extracellular Ca2+ (buffered with 10 mM EGTA) inactivation (by 53 +/- 6 %) and tachyphylaxis (by 42 +/- 7 % across three stimuli) were still observed (n = 8). The same was true when intracellular Ca2+ was buffered by 10 mM BAPTA (inactivation by 49 +/- 4 %, tachyphylaxis by 52 +/- 7 % across three stimuli; n = 13). Thus, inactivation and tachyphylaxis were mainly independent of intra- and extracellular Ca2+. These results indicate that inactivation and tachyphylaxis of heat-evoked inward currents can be observed in vitro, similar to adaptation and suppression of action potential discharges elicited by comparably fast heat stimuli in vivo. Whereas the voltage dependence of I(heat) resembles that of capsaicin-induced membrane currents (I(Caps)), the independence of inactivation and tachyphylaxis of I(heat) from calcium is in clear contrast to I(Caps). A similar difference in calcium dependence of inactivation has been reported between heat-evoked and capsaicin-induced currents through the cloned capsaicin receptor channel VR1. Thus, the properties of I(heat) and of VR1 largely account for the adaptation and suppression of heat-evoked nociceptor discharges. PMID- 11060131 TI - Descending inhibitory reflexes involve P2X receptor-mediated transmission from interneurons to motor neurons in guinea-pig ileum. AB - The role of P2X receptors in descending inhibitory reflexes evoked by distension or mucosal distortion in the guinea-pig ileum was studied using intracellular recording from the circular muscle in a two-chambered organ bath. This allowed separate superfusion of the sites of reflex stimulation and recording, thereby allowing drugs to be selectively applied to different parts of the reflex pathway. Inhibitory junction potentials (IJPs) evoked by electrical field stimulation (EFS) in the recording chamber were compared with those evoked during reflexes to control for effects of P2 receptor antagonists on neuromuscular transmission. The P2 receptor antagonists suramin (100 microM) and pyridoxal phosphate-6-azophenyl-2',4'-disulphonic acid (10 and 60 microM; PPADS), when added to the recording chamber, depressed reflexly evoked IJPs significantly more than those evoked by EFS. In particular, 10 microM PPADS depressed IJPs evoked by distension or mucosal distortion by about 50 %, but had little effect on IJPs evoked by EFS. Blockade of synaptic transmission in the stimulation chamber with a low Ca2+-high Mg2+ solution depressed, but did not abolish, IJPs evoked by distension. The residual reflex IJPs were unaffected by PPADS (10 microM), hyoscine (1 microM), hyoscine plus hexamethonium (200 microM), or hysocine plus hexamethonium plus PPADS in the recording chamber. We conclude that P2X receptors are important for synaptic transmission from descending interneurons to inhibitory motor neurons in descending inhibitory reflex pathways of guinea-pig ileum. Transmission from anally directed axons of distension-sensitive intrinsic sensory neurons to inhibitory motor neurons is unlikely to involve P2X, muscarinic or nicotinic receptors. PMID- 11060132 TI - Functional interdependence of neurons in a single canine intrinsic cardiac ganglionated plexus. AB - To determine the activity characteristics displayed by different subpopulations of neurons in a single intrinsic cardiac ganglionated plexus, the behaviour and co-ordination of activity generated by neurons in two loci of the right atrial ganglionated plexus (RAGP) were evaluated in 16 anaesthetized dogs during basal states as well as in response to increasing inputs from ventricular sensory neurites. These sub-populations of right atrial neurons received afferent inputs from sensory neurites in both ventricles that were responsive to local mechanical stimuli and the nitric oxide donor nitroprusside. Neurons in at least one RAGP locus were activated by epicardial application of veratridine, bradykinin, the beta1-adrenoceptor agonist prenaterol or glutamate. Epicardial application of angiotensin II, the selective beta2-adrenoceptor agonist terbutaline and selective alpha-adrenoceptor agonists elicited inconsistent neuronal responses. The activity generated by both populations of atrial neurons studied over 5 min periods during basal states displayed periodic coupled behaviour (cross correlation coefficients of activities that reached, on average, 0.88 +/- 0.03; range 0.71-1) for 15-30 s periods of time. These periods of coupled activity occurred every 30-50 s during basal states, as well as when neuronal activity was enhanced by chemical activation of their ventricular sensory inputs. These results indicate that neurons throughout one intrinsic cardiac ganglionated plexus receive inputs from mechano- and chemosensory neurites located in both ventricles. That such neurons respond to multiple chemical stimuli, including those liberated from adjacent adrenergic efferent nerve terminals, indicates the complexity of the integrative processing of information that occurs within the intrinsic cardiac nervous system. It is proposed that the interdependent activity displayed by populations of neurons in different regions of one intrinsic cardiac ganglionated plexus, responding as they do to multiple cardiac sensory inputs, forms the basis for integrated regional cardiac control. PMID- 11060133 TI - Characterisation of dark adaptation in human cone pathways: an application of the equivalent background hypothesis. AB - It is well accepted that in rod photoreceptors the photoproducts generated by a bleach cause desensitisation during dark adaptation. We examine whether this notion holds for cones. A model of cone dark adaptation is developed based on the equivalent background concept. The underlying theory of the model relies on a series of assumptions that link psychophysically determined detection thresholds to cone phototransduction. Correction of thresholds for the reduced quantum catching ability of the cones (due to the depletion of photopigment caused by a bleaching light) is an important aspect of the model. Foveal detection thresholds were measured for a small test flash presented on a large steady background field or presented alone after adapting to the background field. Test and background fields were monochromatic, with wavelengths closely matched to promote detection by the luminance mechanism. The model provided a good description of the data collected under these conditions. Parameters of the model were similar for all wavelengths and each observer, as were the derived equivalent background relationships. Analysis of previously published data for Stiles' pi5 mechanism gave analogous results. The model is made up of two components. The early (fast) component is likely to be due to the direct action of the cone equivalent of inactivated Rh* on the G-protein cascade and/or the reverse reaction of the cone equivalent of inactivated Rh* to Rh*. The later (slow) component may be due to the direct action of cone opsin on the G-protein cascade. PMID- 11060134 TI - Action of polysaccharides of similar average mass but differing molecular volume and charge on fluid drainage through synovial interstitium in rabbit knees. AB - Hyaluronan (HA), an anionic polysaccharide of synovial fluid, attenuates fluid loss from joints as joint pressure is raised ('outflow buffering'). The buffering is thought to depend on the expanded molecular domain of the polymer, which causes reflection by synovial extracellular matrix, leading to flow-dependent concentration polarization. We therefore assessed the effects of polysaccharides of differing average molecular volume and charge. Trans-synovial fluid drainage( 8d s) was measured at controlled joint fluid pressure (Pj) in knees of anaesthetized rabbits. The joints were infused with polydisperse HA of weight average mass 2100 kDa (4 mg x ml(-1), n = 17), with polydisperse neutral dextran of similar average mass (2000 kDa; n = 7) or with Ringer solution vehicle (n = 2). The role of polymer charge was assessed by infusions of neutral or sulphated dextran of average molecular mass 500 kDa (n = 6). When HA was present, 8d s increased little with pressure, forming a virtual plateau of approximately 4 microl x min(-1) from 10 to 25 cmH2O. Neutral dextran 2000 failed to replicate this effect. Instead, 8d s increased steeply with Pj, reaching eight times the HA value by 20 cmH2O (P = 0.0001, ANOVA). Dextran 2000 reduced flows in comparison with Ringer solution. Analysis of the aspirated joint fluid showed that 31 +/- 0.07 % (s.e.m.) of dextran 2000 in the filtrand was reflected by synovium, compared with > or = 79 % for HA. The viscometric molecular radius of the dextran, approximately 31 nm, was smaller than that of HA (101-181 nm), as was its osmotic pressure. Anionic dextran 500 failed to buffer fluid drainage, but it reduced fluid escape and synovial conductance d 8d s/dPj more than neutral dextran 500 (P < 0.0001, ANOVA). The anionic charge increased the molecular volume and viscosity of dextran 500. The results support the hypothesis that polymer molecular volume influences its reflection by interstitial matrix and outflow buffering. Polymer charge influences flow through an effect on viscosity and possibly electrostatic interactions with negatively charged interstitial matrix. PMID- 11060135 TI - Effects of betamethasone administration to the fetal sheep in late gestation on fetal cerebral blood flow. AB - Glucocorticoid administration to women at risk of preterm delivery to accelerate fetal lung maturation has become standard practice. Antenatal glucocorticoids decrease the incidence of intraventricular haemorrhage as well as accelerating fetal lung maturation. Little is known regarding side effects on fetal cerebral function. Cortisol and synthetic glucocorticoids such as betamethasone increase fetal blood pressure and femoral vascular resistance in sheep. We determined the effects of antenatal glucocorticoid administration on cerebral blood flow (CBF) in fetal sheep. Vehicle (n = 8) or betamethasone (n = 8) was infused over 48 h via the jugular vein of chronically instrumented fetal sheep at 128 days gestation (term 146 days). The betamethasone infusion rate was that previously shown to produce fetal plasma betamethasone concentrations similar to human umbilical vein concentrations during antenatal glucocorticoid therapy. Regional CBF was measured in 10 brain regions, using coloured microspheres, before and 24 and 48 h after onset of treatment, and during hypercapnic challenges performed before and 48 h after onset of betamethasone exposure. Betamethasone exposure decreased CBF in all brain regions measured except the hippocampus after 24 h of infusion (P < 0.05). The CBF decrease was most pronounced in the thalamus and hindbrain (45-50% decrease) and least pronounced in the cortical regions (35-40% decrease). It was mediated by an increase in cerebral vascular resistance (CVR, P < 0.05) and led to a decrease in oxygen delivery to subcortical and hindbrain structures of 30-40%, to 8.6 +/- 1.1 ml x (100 g)(-1) x min(-1), and 40-45 %, to 11.0 +/- 1.6 ml x 100 g(-1) x min(-1), respectively (P < 0.05). After 48 h of betamethasone treatment, the reduction in CBF was diminished to about 25-30 %, but was still significant in comparison to vehicle-treated fetuses in all brain regions except three of the five measured cortical regions (P < 0.05). CVR and oxygen delivery were unchanged in comparison to values at 24 h of treatment. The CBF increase in response to hypercapnia was diminished (P < 0.05). These observations demonstrate for the first time that glucocorticoids exert major vasoconstrictor effects on fetal CBF. This mechanism may protect the fetus against intraventricular haemorrhage both at rest and when the fetus is challenged. Betamethasone exposure decreased the hypercapnia-induced increase in CBF (P < 0.05) due to decreased cerebral vasodilatation (P < 0.05). PMID- 11060136 TI - Time-varying changes in corticospinal excitability accompanying the triphasic EMG pattern in humans. AB - Nine healthy subjects performed single rapid wrist movements from neutral to targets at 20 deg of flexion or extension in response to an auditory cue. Surface EMG was recorded from the wrist flexors and extensors together with wrist position. Movements in both directions were characterised by the usual triphasic pattern of EMG activity in agonist (AG1), antagonist (ANTAG) and again in agonist (AG2) muscles. Single pulses of transcranial magnetic stimulation (TMS) were applied over the motor cortex at an intensity of 80 % of resting threshold at random times between 80 and 380 ms after the cue. We measured the peak-to-peak amplitude of the evoked motor potential (MEP) and the integrated EMG (IEMG) activity that preceded the MEP. In a separate set of experiments H reflexes were elicited in the wrist flexors instead of MEPs. MEP amplitudes in the agonist muscle increased by an average of 10 +/- 8 ms (range -1 to 23 ms) prior to the onset of the AG1 burst and were associated with an increase of over sevenfold in the MEP:IEMG ratio, irrespective of movement direction. Agonist H reflex amplitudes were linearly related to, and increased at the same time as, changes in agonist IEMG. The principal ANTAG burst was not preceded by an increase in the antagonist muscle MEP:IEMG ratio. No relationship was found between the amplitude of the antagonist H reflexes and the preceding antagonist IEMG. Five subjects showed an increase in the MEP:IEMG ratio preceding and during the initial part of the AG2 burst. Our method of analysis shows that changes in motor cortical excitability mediating the initiation of movement occur much closer to the onset of EMG activity (less than 23 ms) than the 80-100 ms lead time previously reported. The lack of such changes before the onset of the ANTAG burst suggests that this may be initiated by a different, perhaps subcortical, mechanism. PMID- 11060137 TI - Effect of prolonged, submaximal exercise and carbohydrate ingestion on monocyte intracellular cytokine production in humans. AB - The present study was undertaken to examine the effect of exercise and carbohydrate (CHO) ingestion on intracellular monocyte cytokine production. Subjects performed 2 h of cycling at 70 % peak pulmonary O2 uptake (VO2,peak) while ingesting either an 8 % CHO beverage or a sweet placebo. Whole blood was incubated with (stimulated) or without (spontaneous) lipopolysaccharide (LPS) and surface stained for monocyte surface antigens. The cells were permeabilised, stained for intracellular cytokines and analysed using flow cytometry. Exercise had no effect on the number of monocytes spontaneously producing cytokines, but the number of stimulated IL-1alpha-, TNF-alpha- and IL-6-positive monocytes were elevated (P < 0.01) immediately post-exercise and 2 h post-exercise. These stimulated cells produced less (P < 0.05) TNF-alpha immediately post-exercise, and less (P < 0.05) TNF-alpha and IL-1alpha 2 h post-exercise. There was a small, but significant increase (P < 0.05) in the plasma IL-6 concentration immediately post-exercise. Exercise resulted in an elevation (P < 0.01) in the plasma adrenaline concentration in the placebo trial, and ingestion of CHO attenuated this increase. CHO ingestion had no effect on monocyte cytokine production, plasma IL-6 or circulating leukocyte numbers. These data suggest that circulating monocytes are not the origin of increased levels of plasma IL-6 during exercise: prolonged cycling exercise increased the number of monocytes producing cytokines upon stimulation, but these cells produced less cytokines post-exercise. In addition, attenuation of plasma adrenaline levels had no effect on plasma IL-6 or monocyte cytokine production. PMID- 11060138 TI - The role of gravity in human walking: pendular energy exchange, external work and optimal speed. AB - During walking on Earth, at 1.0 g of gravity, the work done by the muscles to maintain the motion of the centre of mass of the body (W(ext)) is reduced by a pendulum-like exchange between gravitational potential energy and kinetic energy. The weight-specific W(ext) per unit distance attains a minimum of 0.3 J x kg(-1) x m(-1) at about 4.5 km x h(-1) in adults. The effect of a gravity change has been studied during walking on a force platform fixed to the floor of an aircraft undergoing flight profiles which resulted in a simulated gravity of 0.4 and 1.5 times that on Earth. At 0.4 g, such as on Mars, the minimum W(ext) was 0.15 J x kg(-1) x m(-1), half that on Earth and occurred at a slower speed, about 2.5 km x h(-1). The range of walking speeds is about half that on Earth. At 1.5 g, the lowest value of W(ext) was 0.60 J x kg(-1) x m(-1), twice that on Earth; it was nearly constant up to about 4.3 km x h(-1) and then increased with speed. The range of walking speeds is probably greater than that on Earth. A model is presented in which the speed for an optimum exchange between potential and kinetic energy, the 'optimal speed', is predicted by the balance between the forward deceleration due to the lift of the body against gravity and the forward deceleration due to the impact against the ground. In conclusion, over the range studied, gravity increases the work required to walk, but it also increases the range of walking speeds. PMID- 11060139 TI - Adipose tissue, the anatomists' Cinderella, goes to the ball at last, and meets some influential partners. PMID- 11060140 TI - Management of leg ulcers. AB - Leg ulcer is a leading cause of morbidity among older subjects, especially women in the Western world. About 400 years BC, Hippocrates wrote, "In case of an ulcer, it is not expedient to stand, especially if the ulcer be situated on the leg". Hippocrates himself had a leg ulcer. The best treatment of any leg ulcer depends upon the accurate diagnosis and the underlying aetiology. The majority of leg ulcers are due to venous disease and/or arterial disease, but the treatment of the underlying cause is far more important than the choice of dressing. The aetiology, pathogenesis, treatment, and the future trends in the management of the leg ulcers are discussed in this review. PMID- 11060141 TI - Pregnancy and the lungs. PMID- 11060142 TI - Unknown primary tumours. AB - Unknown primary tumours (UPTs) are defined by the absence of a primary tumour in biopsy proved metastatic cancer. These tumours have a specific biology with clinical characteristics of rapid progression and random atypical metastases. Cytogenetic abnormalities have been demonstrated, particularly deletion of chromosome 1p. Diagnostic evaluation that includes pathology review, physical examination, chest radiography, computed tomography of the abdomen, and mammography is directed at the identification of treatable subsets. Based on clinicopathological criteria, therapy responsive subsets of patients with UPTs can be defined. These subsets have a better prognosis than the average median survival time of four months in patients with UPTs. PMID- 11060143 TI - Influence of smoking on asthmatic symptoms and allergen sensitisation in early childhood. AB - BACKGROUND: Apart from heredity, several early life environmental factors are implicated in the development of childhood asthma. Maternal smoking is believed to increase asthmatic symptoms but its influence on the development of allergen sensitisation is debatable. STUDY DESIGN: A whole population birth cohort was reviewed at ages 1, 2, and 4 years. Of 1218 children seen at 4 years, 981 (80.5%) were skin prick tested with a battery of common food and aeroallergens. Smoking history was recorded at birth and updated at each follow up and its impact on the development of asthma and allergen sensitisation in the children was assessed. RESULTS: Two hundred and fifty mothers smoked during pregnancy (20.5%) and 307 (25.2%) after childbirth. Maternal smoking in pregnancy was associated with low birth weight (mean (SD): 3.3 (0.5) v. 3.5 (0.5) kg; p<0.001). Smoking mothers were more often from lower social classes (31.8% v. 16%, p<0. 001) and they breast fed their babies for a shorter duration (8.5 (11.4) v. 16.6 (15.2) weeks; p<0.001). The difference in breast feeding duration was partly due to a higher proportion of smoking mothers who never breast fed their babies. Although at age 2 years asthmatic symptoms were associated with exposure to maternal tobacco smoke (odds ratio 2.2, 95% confidence interval 1.5 to 3.4; p<0.001), this association was lost by 4 years. However, maternal smoking was a significant risk factor in a subgroup of children with asthmatic symptoms but negative skin prick test. Maternal smoking did not increase allergen sensitisation at age 4 years. No effect of paternal smoking on asthma was observed in the children. PMID- 11060144 TI - Gallstones and cholecystectomy in modern Britain. AB - BACKGROUND: It has been suggested that gallstone disease is now commoner, and that this might explain an increase in cholecystectomy rates, though conclusive evidence has been lacking. METHODS: All the non-forensic necropsy results for Dundee 1953-98 were examined to assess the prevalence of gallstone disease. The NHS Scotland annual cholecystectomy figures were extracted from their earliest availability in 1961 up to the present. The subgroup of patients from Dundee was analysed separately, as were laparoscopic procedures, which were recorded from 1991. RESULTS: Gallstone disease was much commoner in 1974-98 than in 1953-73. Increasing age was the main determinant of gallstone disease. Though gallstone disease was commoner in women than men aged 40-89, there was no sex difference under 40 or over 90 years. Cholecystectomy became much commoner in the 1960s when frequency of gallstone disease did not change. It increased further in the 1970s, peaking in 1977-8. There was a gradual fall in rates in the 1980s when gallstone prevalence remained high. There was a further moderate rise in the 1990s after the wide introduction of laparoscopic cholecystectomy. Cholecystectomy is now much commoner in young women and this change started in the 1960s. By contrast, cholecystectomy in men has become more prevalent in the older age group. CONCLUSIONS: Gallstones were definitely more common in both sexes at all ages over 40 in the last 25 years. Changes in the cholecystectomy rates are only partly explained by changes in gallstone prevalence, and are more determined by surgical practice. PMID- 11060145 TI - Disseminated fungal infection complicated with pulmonary haemorrhage in a case of acute myeloid leukaemia. AB - Pulmonary haemorrhage is a common necropsy finding in acute leukaemia, however, it is rarely diagnosed during life. A man with acute myeloid leukaemia is reported who presented with disseminated fungal infection, anaemia, thrombocytopenia, and subconjuctival and petechial haemorrhages. During the course of the patient's illness, the chest infection was complicated with bilateral pulmonary haemorrhage. The diagnosis of pulmonary haemorrhage was based on characteristic clinical and radiological findings. The patient improved on treatment. PMID- 11060146 TI - Primary haematogenous osteomyelitis of the patella: a rare cause for anterior knee pain in an adult. AB - Acute osteomyelitis of the patella is a very rare condition, which commonly affects children between the ages of 5-15 years. Primary haematogenous osteomyelitis in an adult usually occurs in patients with associated risk factors like intravenous drug abuse, HIV infection, and trauma. This report discusses a similar condition in a 46 year old women with no associated predisposing risk factors. The rarity of this condition and its atypical presentation should be borne in mind while treating an adult patient with anterior knee pain. Point tenderness over the patella should alert a physician to the possibility of osteomyelitis of the patella. The value of bone scan and computed tomography in the early stages to help diagnose this condition has been stressed. The literature has been reviewed and discussed briefly. PMID- 11060147 TI - Neuralgic amyotrophy as a presenting feature of infective endocarditis. AB - A 35 year old man presented to his general practitioner with severe right shoulder pain and subsequent weakness and wasting of the muscles in the affected shoulder girdle three weeks after a dental filling. His symptoms persisted despite standard treatment. He developed malaise, night sweats, weight loss, a petechial rash and a microcytic anaemia. On admission to hospital three months after the start of his symptoms he had also developed splenomegaly and the murmur of aortic regurgitation. Investigations confirmed the diagnoses of infective endocarditis and neuralgic amyotrophy. In this case neuralgic amyotrophy appears to have been the presenting feature of infective endocarditis. This association has not previously been described. PMID- 11060148 TI - Dental anomalies in Williams syndrome. PMID- 11060149 TI - Sweet's syndrome in association with Crohn's disease. AB - A case of Sweet's syndrome in association with Crohn's disease in a young woman is reported. Sweet's syndrome is a rare extraintestinal manifestation of Crohn's disease and ulcerative colitis. PMID- 11060150 TI - A case of digital vasculitis. PMID- 11060151 TI - Abdominal wall thickening in a middle aged man. PMID- 11060152 TI - A 71 year old man with a facial lesion. PMID- 11060153 TI - The pale and limping child. PMID- 11060154 TI - Rigidity, hyperthermia, and altered mental status. PMID- 11060155 TI - A rare cause of recurrent melaena in an elderly women. PMID- 11060156 TI - Abdominal lump in an infertile man. PMID- 11060157 TI - A vanishing pituitary mass. PMID- 11060158 TI - Shortness of breath and diffuse chest pain. PMID- 11060159 TI - An elderly man with muscle cramps. PMID- 11060161 TI - A case of digital vasculitis PMID- 11060160 TI - Left kidney mass in a 45 year old woman. PMID- 11060162 TI - Abdominal wall thickening in a middle aged man PMID- 11060163 TI - A 71 year old man with a facial lesion PMID- 11060165 TI - Rigidity, hyperthermia, and altered mental status PMID- 11060164 TI - The pale and limping child PMID- 11060166 TI - A rare cause of recurrent meleana in an elderly women PMID- 11060167 TI - Abdominal lump in an infertile man PMID- 11060168 TI - A vanishing pituitary mass PMID- 11060169 TI - Shortness of breath and diffuse chest pain PMID- 11060171 TI - Left kidney mass in a 45 year old woman PMID- 11060170 TI - An elderly man with muscle cramps PMID- 11060172 TI - The real cost of aspirin. AB - Aspirin is a widely used drug and perceived by most physicians to be inexpensive. High rates of concurrent gastroprotective agents are reported from a study of cardiology outpatients. Aspirin takers are more likely to also be taking a proton pump inhibitor, H(2) antagonist, or antacid than non-aspirin takers. They are more than 10 times as likely to be experiencing upper gastrointestinal symptoms. Although aspirin is inexpensive, it is emphasised that the overall cost implications for therapy can be significant and it is suggested that it may be more appropriate to consider the use of alternative antiplatelet agents in patients who tolerate aspirin poorly. PMID- 11060173 TI - Indomethacin induced psychosis. AB - Indomethacin is a commonly prescribed non-steroidal anti-inflammatory drug. While its adverse effects on gastrointestinal and renal systems are well described, its central nervous system effects are less well known. This case report describes an elderly man, prescribed indomethacin for gout, who presented with psychosis. PMID- 11060174 TI - Perceptions of malaria transmission before Ross' discovery in 1897. PMID- 11060175 TI - Evidence and clinical medicine PMID- 11060177 TI - Diagnosing cancer in primary care PMID- 11060176 TI - Evidence-based clinical practice-concepts & approaches PMID- 11060178 TI - A colour handbook of gastroenterology PMID- 11060180 TI - Davies textbook of adverse drug reactions PMID- 11060179 TI - ABC of sexual health PMID- 11060181 TI - Overuse and underuse of colonoscopy in a European primary care setting. AB - BACKGROUND: Efforts to decrease overuse of health care may result in underuse. Overuse and underuse of colonoscopy have never been simultaneously evaluated in the same patient population. METHODS: In this prospective observational study, the appropriateness and necessity of referral for colonoscopy were evaluated by using explicit criteria developed by a standardized expert panel method. Inappropriate referrals constituted overuse. Patients with necessary colonoscopy indications who were not referred constituted underuse. Consecutive ambulatory patients with lower gastrointestinal (GI) symptoms from 22 general practices in Switzerland, a country with ready access to colonoscopy, were enrolled during a 4 week period. Follow-up data were obtained at 3 months for patients who did not undergo a necessary colonoscopy. RESULTS: Eight thousand seven hundred sixty patient visits were screened for inclusion; 651 patients (7.4%) had lower GI symptoms (mean age 56.4 years, 68% women). Of these, 78 (12%) were referred for colonoscopy. Indications for colonoscopy in 11 patients (14% of colonoscopy referrals or 1.7% of all patients with lower GI symptoms) were judged inappropriate. Among 573 patients not referred for the procedure, underuse ranged between 11% and 28% of all patients with lower GI symptoms, depending on the criteria used. CONCLUSIONS: Applying criteria from an expert panel of nationally recognized experts indicates that underuse of referral for colonoscopy exceeds overuse in primary care in Switzerland. To improve quality of care, both overuse and underuse of important procedures must be addressed. PMID- 11060182 TI - Factors affecting insertion time and patient discomfort during colonoscopy. AB - BACKGROUND: Successful colonoscopy depends on insertion of the instrument to the cecum, precise observation, and minimal patient discomfort during the procedure. The aim of this prospective study was to determine whether certain variables are associated with insertion time and patient discomfort during colonoscopy. METHODS: Nine hundred nine consecutive colonoscopic examinations performed by a single endoscopist in patients without obstructive disease of the colorectum were analyzed. Four liters of Colonlyte (Taejun, Seoul, Korea) were used for bowel cleansing, and meperidine (25 mg) was administered intramuscularly 10 minutes before the procedure. The degree of patient discomfort was assessed using a 5 level Likert scale. RESULTS: Among 909 study patients, colonoscopy was completed to the cecum in 876 patients (96.4%). The adjusted completion rate was 98% and mean insertion time for complete colonoscopy was 6.9+/-4.2 minutes. Colonoscopy caused less patient discomfort than barium enema or esophagogastroduodenoscopy. Multivariate logistic regression analysis demonstrated that inadequate bowel cleansing, advanced age, and constipation as an indication are independent factors associated with prolonged insertion time (>10 minutes). Female gender was the only independent factor associated with significant discomfort (> or = level 4) during colonoscopy. CONCLUSIONS: Among the factors affecting insertion time and patient discomfort during colonoscopy, unsatisfactory bowel preparation was the only correctable factor. PMID- 11060183 TI - Patient tolerance of colonoscopy without sedation during screening examination for colorectal polyps. AB - BACKGROUND: The administration of sedative drugs at colonoscopy has its drawbacks such as increases in the rate of complications and cost. Our aim was to study how individuals, drawn from a population registry and invited to undergo screening colonoscopy for colorectal polyps, experienced the procedure without conscious sedation. METHODS: Four hundred fifty-one individuals underwent the screening examination (median age 67 years, range 63 to 72). The cecum was intubated in 369 (82%). Fourteen days after the examination, 429 of the attendees received a questionnaire designed to evaluate their tolerance of the procedure. RESULTS: Four hundred nine participants (95%) replied.Twenty-one (5%) of these individuals found the examination very uncomfortable, 184 (45%) found it moderately uncomfortable, and 204 (50%) did not find it uncomfortable. A larger proportion of women than men, 110 (63%) versus 79 (41%), found the procedure very or moderately uncomfortable (p<0.001). Three hundred sixty-eight (90%) individuals stated that they would undergo repeat colonoscopy in 5 years. CONCLUSION: In this screening setting, routine use of conscious sedation did not seem to be necessary, as most participants found the examination to be only moderately uncomfortable or not at all uncomfortable. Colonoscopy without conscious sedation may, however, reduce the rate of intubation of the cecum and increase the risk of missing adenomas and cancers. PMID- 11060184 TI - EUS changes predictive for recurrence of esophageal varices in patients treated by combined endoscopic ligation and sclerotherapy. AB - BACKGROUND: Recurrence of varices is still common after endoscopic treatment of esophageal varices. In this study, predictive signs of variceal recurrence were investigated by ultrasonic (US) miniature probe in patients treated by combined endoscopic ligation and sclerotherapy. METHODS: Detectability of vessels by US miniature probe was evaluated first in rats. In 41 patients treated by combined therapy, the esophagus and the cardia region were examined by US miniature probe. In 25 patients examined by percutaneous transhepatic portography, the relationship between US miniature probe and percutaneous transhepatic portography findings was evaluated. RESULTS: The smallest vessel detected by US miniature probe was 0.3 mm in diameter in the study using intra-abdominal vessels of rat. After variceal eradication, US miniature probe showed intramural vessels in the cardia that were classified as follows: grade I, a few vessels (19 patients, 46%); grade II, uniformly scattered vessels (11, 27%); grade III, abundant vessels resembling a honeycomb (11, 27%). As the sonographic grade increased, the rate of variceal recurrence increased. As the venographic grade of staining in the distal esophagus increased, the esophageal wall became thicker and the sonographic grade at the cardia increased. CONCLUSIONS: Endosonographic evaluation of the distal esophagus and cardia is predictive of variceal recurrence. PMID- 11060185 TI - Sphincter of Oddi contractile function after balloon dilation: detailed manometric evaluation in conscious dogs. AB - BACKGROUND: Endoscopic balloon dilation is under investigation as a treatment modality for bile duct stones. It may have an advantage of preserving the sphincter of Oddi function, but little is known about sphincter of Oddi cyclic motility after dilation. METHODS: Four dogs with a duodenal cannula underwent sphincter of Oddi dilation and repeated manometry to assess sphincter of Oddi cyclic motility until 3 months after dilation. Histologic changes in the sphincter of Oddi were examined in another group of four dogs. RESULTS: Motility index (sum of amplitude of sphincter of Oddi phasic waves counted per minute) and basal pressure decreased on day 3. Sphincter of Oddi amplitude during phase III of the duodenal migrating motor complex tended to be increased on day 3 and decreased to the minimum on day 21. Thereafter, it gradually recovered to baseline. By histology, severe acute inflammation was present in the sphincter of Oddi muscle layer on day 3. However, basal pressure remained significantly low even 3 months after dilation. CONCLUSIONS: Sphincter of Oddi amplitude is incompletely reduced on day 3 after balloon dilation. Sphincter of Oddi basal pressure and motility index in the early phase of sphincter of Oddi cyclic motility remain low for at least 3 months after dilation. Further long-term follow-up is necessary to determine whether sphincter of Oddi function is actually preserved. PMID- 11060186 TI - Influence of cholangiography on biliary sphincter of Oddi manometric parameters. AB - BACKGROUND: When sphincter of Oddi manometry (SOM) and endoscopic retrograde cholangiopancreatography are performed at the same session, SOM is usually performed immediately before ductography because of concern about the accuracy of the manometric recording after contrast medium injection. However, it would be preferable to inject contrast medium first to identify other causes for a patient's symptoms, allowing selective use of SOM. The aim of this study was to evaluate the effect of cholangiography on sphincter of Oddi (SO) basal pressure. METHODS: Twenty-five patients with suspected SO dysfunction were prospectively studied. Conventional station pull-through manometry of the biliary part of the sphincter was performed before and after cholangiography. The intraductal pressure and basal sphincter pressure were evaluated. RESULTS: The mean intraductal pressure was 8+/-5.5 mm Hg before and 13.3+/-6.8 mm Hg after contrast medium injection (p< 0.01). However, the basal sphincter pressure was not significantly altered (52.9+/-42.1 mm Hg vs. 55.1+/-38.1 mm Hg, p = 0.52). Concordance (normal vs. abnormal) between the basal sphincter pressure before and after ductography was seen in 24 of 25 patients (96%). CONCLUSIONS: Intraductal installation of contrast medium immediately before SOM infrequently alters SO basal pressure in a clinically significant manner. We therefore believe that this sequence can be utilized in clinical practice. PMID- 11060187 TI - Cholangioscopic findings in bile duct tumors. AB - BACKGROUND: Cholangioscopy has been used in the treatment of bile duct stones and the diagnosis of various bile duct tumors. However, the cholangioscopic characteristics of the various types of bile duct tumors have not been clearly described. We analyzed the results of cholangioscopic examinations and classified the findings according to tumor histology. METHODS: Cholangioscopic findings from 111 patients with benign or malignant bile duct tumors were reviewed. The mucosal changes, the presence of neovascularization, and the patterns of luminal narrowing were analyzed and compared with the histologic diagnosis. RESULTS: Bile duct adenocarcinoma can be classified into 3 different types according to the cholangioscopic findings: nodular, papillary, and infiltrative. Bile duct adenoma, hepatocellular carcinoma and other types of bile duct cancer such as mucin-hypersecreting cholangiocarcinoma, biliary cystadenocarcinoma, and squamous cell carcinoma also presented unique cholangioscopic characteristics. CONCLUSIONS: Bile duct tumors exhibit characteristic cholangioscopic findings and cholangioscopy seems to be useful for differential diagnosis. PMID- 11060188 TI - Tumor vessel: a valuable cholangioscopic clue of malignant biliary stricture. AB - BACKGROUND: An irregularly dilated and tortuous vessel, the so-called tumor vessel, is considered to be one of the cholangioscopic features that suggest biliary malignancy. This is a prospective analysis of the presence of a tumor vessel as a finding that discriminates between benign and malignant biliary strictures. METHODS: From August 1997 to August 1998, a total of 63 patients with biliary strictures diagnosed with endoscopic retrograde cholangiography or percutaneous transhepatic cholangiography obtained during percutaneous transhepatic biliary drainage tube placement were included in this study. Strictures were characterized as benign or malignant based on the observation of tumor vessels. The results were compared with those of percutaneous transhepatic cholangiography-guided biopsy and final diagnosis. RESULTS: Forty-one patients were confirmed to have malignant strictures and 22 had benign biliary strictures. Cancer was confirmed by histopathologic evaluation of biopsies in 33 of 41 patients with malignancy (80.4%). Tumor vessel was seen in 25 of 41 patients with malignancy (61%). No patients with benign stricture had tumor vessels. Of the 8 patients with negative percutaneous transhepatic cholangioscopy-guided biopsies but with a final diagnosis of malignancy, 6 had tumor vessels. Combining the observation of tumor vessel and percutaneous transhepatic cholangiography-guided biopsy resulted in a diagnosis of malignancy in 39 of 41 patients (96%) and significantly increased the rate of preoperative diagnosis when compared with percutaneous transhepatic cholangiography-guided biopsy or presence of tumor vessel alone (p<0.05). CONCLUSION: The presence of tumor vessel may be a valuable cholangioscopic finding that indicates the presence of a malignant biliary stricture. The combination of tumor vessel observation and percutaneous transhepatic cholangiography-guided biopsy may improve the preoperative diagnosis of malignancy. PMID- 11060189 TI - Long-term follow-up after treatment of Mirizzi syndrome by peroral cholangioscopy. AB - BACKGROUND: The standard treatment for Mirizzi syndrome is surgical, although endoscopic and percutaneous management have also been described. The aim of this study was to evaluate the usefulness of shock wave lithotripsy combined with peroral cholangioscopy and its long-term outcome in patients with Mirizzi syndrome. METHODS: The records of 25 patients with Mirizzi syndrome who underwent endoscopic treatment between April 1990 and November 1998 were retrospectively reviewed. Shock wave lithotripsy was performed under direct vision with a "mother baby" endoscope system in 2 patients with type I and 23 with type II Mirizzi syndrome (12 men and 13 women, mean age 60 years). Follow-up data were obtained from clinical records or through telephone interviews. RESULTS: In the two patients with type I, the cholangioscopic approach failed and both patients underwent open cholecystectomy. The 23 patients with type II were all successfully treated with shock wave lithotripsy alone. The cholangioscopic approach was unsuccessful in the treatment of residual gallbladder stones. Follow up data were obtained in all but one patient (mean 43.6 months, range 4 to 103 months). Of the 23 patients with type II, 12 with no gallbladder stones had remained asymptomatic during the follow-up period. Of the 6 patients with type II with large residual gallbladder stones, 4 had acute cholangitis due to stone migration 6, 9, 28, and 34 months after endoscopic treatment. Two patients died during the follow-up period, one of non-biliary causes and the other of coexistent gallbladder carcinoma. CONCLUSIONS: Endoscopic treatment of Mirizzi syndrome using peroral cholangioscopy is a safe and effective alternative to surgery, especially in patients with the type II syndrome. A favorable long-term outcome depends on the absence of large residual gallbladder stones. PMID- 11060190 TI - Safety and usefulness of percutaneous transhepatic cholecystoscopy examination in high-risk surgical patients with acute cholecystitis. AB - BACKGROUND: The aim of the present study was to evaluate the diagnostic and therapeutic usefulness of percutaneous transhepatic cholecystoscopy in high-risk surgical patients with acute cholecystitis. METHODS: Between January 1992 and June 1998, there were 33 consecutive patients who underwent percutaneous transhepatic cholecystostomy and subsequent percutaneous transhepatic cholecystoscopy for the management of acute cholecystitis. RESULTS: Percutaneous transhepatic cholecystostomy and subsequent percutaneous transhepatic cholecystoscopy were successfully accomplished in all 33 patients. During percutaneous transhepatic cholecystoscopy, minor complications (2 episodes of minor bleeding during electrohydraulic lithotripsy, 2 of tube dislodgement, and 1 of bile leakage to peritoneum) occurred in 5 patients. Percutaneous transhepatic cholecystoscopy revealed gallstones in 26 cases, sludge ball in 3, gallbladder carcinoma in 3, and 1 case of clonorchiasis related with acute cholecystitis. The 3 gallbladder cancers which were not identified radiologically were found incidentally during percutaneous transhepatic cholecystoscopy. For the 26 patients with gallstones, percutaneous transhepatic cholecystoscopy and concomitant stone removal were successful in 1 to 4 consecutive sessions (mean 2.2 sessions). Gallstones recurred in 3 of 22 patients (14%) during the mean follow-up period of 27 months. All of them remain asymptomatic. CONCLUSION: Percutaneous transhepatic cholecystostomy may be justified in the management of acute cholecystitis in selected patients with high surgical risk. PMID- 11060191 TI - Endoscopic assessment of hiatal hernia repair. PMID- 11060192 TI - Esophageal squamous papilloma causing dysphagia. PMID- 11060193 TI - Hepatocellular carcinoma invading bile duct. PMID- 11060194 TI - Endoscopic hemoclip placement for post-sphincterotomy bleeding. PMID- 11060195 TI - Endoscopic valvuloplasty for GERD. AB - BACKGROUND: The transoral, endoscopic route has been suggested as a possible approach for the correction of severe gastroesophageal reflux. Such a procedure would involve no mobilization of the cardia or other structures. The optimal placement, number, and configuration of sutures remains undefined. METHODS: With the use of a previously developed endoscopic sewing machine, this study was undertaken in baboons with two suture arrangements immediately below the lower esophageal sphincter. A linear arrangement (group I) and a circular arrangement (group II) were compared. During the 6 months after the procedure, the animals were evaluated using manometry, fluoroscopic barium swallow, upper gastrointestinal endoscopy, and a pressure volume test. RESULTS: A significant increase in lower esophageal sphincter length was demonstrated only in group II (p = 0. 010). A significant increase in lower esophageal sphincter pressure was demonstrated only in group I animals (p = 0.008). The abdominal length increased in group I (p = 0.004) and group II (p = 0.004). The yield pressure and yield volume did not differ significantly from those measured previously in control animals. No evidence of reflux, stricture formation, esophagitis, or other pathology was noted. CONCLUSIONS: Some manometric parameters associated with gastroesophageal reflux are altered by the endoscopic placement of sutures below the gastroesophageal junction, with no associated serious complications. PMID- 11060196 TI - EUS imaging of the arteria lusoria: case series and review. AB - BACKGROUND: The arteria lusoria is an aberrant right subclavian artery that passes dorsally between the esophagus and spine after branching off from the aortic arch. The role of endoscopic ultrasonography (EUS) in the diagnosis of the arteria lusoria has not been established. The possibility of demonstrating this vascular anomaly with EUS and estimating its prevalence was assessed. METHODS: From December 1991 to September 1998, EUS of the upper gastrointestinal tract was performed in 3334 consecutive patients for various reasons. After imaging of the target organ(s), the echoendoscope was slowly pulled back while imaging the mediastinum proximally to the superior aspect of the aortic arch. In every patient, the area of the aortic arch was carefully inspected and special attention was given to vessels crossing from left to right between the esophagus and spine. RESULTS: During the study period, an arteria lusoria was discovered in 12 of 3334 patients (0.36%: 95% CI [0.16%, 0.56%]). There were 6 men and 6 women; mean age was 64+/-14.7 years. In all 12 patients the arteria lusoria passed between the esophagus and spine. In none of the patients could symptoms be attributed to the presence of this anatomic variant. CONCLUSIONS: An arteria lusoria can be detected with upper gastrointestinal EUS and was found in 0.36% of patients. EUS can accurately demonstrate this vascular anomaly. PMID- 11060197 TI - Solitary duodenal ulcer: a new presentation of eosinophilic gastroenteritis. PMID- 11060198 TI - Metastatic breast cancer causing jaundice by a unique mechanism. PMID- 11060199 TI - Colonoscopy in flowing water for lower GI bleeding: a reliable method for confirmation of bleeding points for endoscopic treatment. PMID- 11060200 TI - Lift and ligate: a new technique to treat a bleeding polypectomy stump. PMID- 11060201 TI - Squamous metaplasia of the rectum after argon plasma coagulation. PMID- 11060202 TI - EUS diagnosis of lymphomatous polyposis. PMID- 11060203 TI - Endoscopic treatment of chronic pancreatitis in a case of "horseshoe" pancreas. PMID- 11060204 TI - Pyopneumopericardium due to an esophagopericardial fistula: treatment with a coated expandable metal stent. PMID- 11060206 TI - Is referral for colonoscopy underutilized by primary care physicians? PMID- 11060205 TI - Methemoglobinemia related to topical benzocaine use: is it time to reconsider the empiric use of topical anesthesia before sedated EGD? PMID- 11060208 TI - Endoscopy simulators: the state of the art, 2000. PMID- 11060207 TI - The societal perspective: monocular or bifocal? Presidential address. PMID- 11060209 TI - Comment PMID- 11060210 TI - Complications of outpatient colonoscopy. PMID- 11060211 TI - Response PMID- 11060212 TI - Grading ERCPs by degree of difficulty: a great concept. PMID- 11060213 TI - Genetic heterogeneity of Usher syndrome: analysis of 151 families with Usher type I. AB - Usher syndrome type I is an autosomal recessive disorder marked by hearing loss, vestibular areflexia, and retinitis pigmentosa. Six Usher I genetic subtypes at loci USH1A-USH1F have been reported. The MYO7A gene is responsible for USH1B, the most common subtype. In our analysis, 151 families with Usher I were screened by linkage and mutation analysis. MYO7A mutations were identified in 64 families with Usher I. Of the remaining 87 families, who were negative for MYO7A mutations, 54 were informative for linkage analysis and were screened with the remaining USH1 loci markers. Results of linkage and heterogeneity analyses showed no evidence of Usher types Ia or Ie. However, one maximum LOD score was observed lying within the USH1D region. Two lesser peak LOD scores were observed outside and between the putative regions for USH1D and USH1F, on chromosome 10. A HOMOG chi(2)((1)) plot shows evidence of heterogeneity across the USH1D, USH1F, and intervening regions. These results provide conclusive evidence that the second most-common subtype of Usher I is due to genes on chromosome 10, and they confirm the existence of one Usher I gene in the previously defined USH1D region, as well as providing evidence for a second, and possibly a third, gene in the 10p/q region. PMID- 11060214 TI - Colour constancy in the swallowtail butterfly Papilio xuthus. AB - We have recently shown that the Japanese yellow swallowtail butterfly Papilio xuthus uses colour vision when searching for food. In the field, these butterflies feed on nectar provided by flowers of various colours not only in direct sunlight but also in shaded places and on cloudy days, suggesting that they have colour constancy. Here, we tested this hypothesis. We trained newly emerged Papilio xuthus to feed on sucrose solution on a paper patch of a certain colour under white illumination. The butterflies were then tested under both white and coloured illumination. Under white illumination, yellow- and red trained butterflies selected the correctly coloured patch from a four-colour pattern and from a colour Mondrian collage. Under four different colours of illumination, we obtained results that were fundamentally similar to those under white illumination. Moreover, we performed critical tests using sets of two similar colours, which were also correctly discriminated by trained butterflies under coloured illumination. Taken together, we conclude that the butterfly Papilio xuthus exhibits some degree of colour constancy when searching for food. PMID- 11060215 TI - The transporter-like protein inebriated mediates hyperosmotic stimuli through intracellular signaling. AB - We cloned the inebriated homologue MasIne from Manduca sexta and expressed it in Xenopus laevis oocytes. MasIne is homologous to neurotransmitter transporters but no transport was observed with a number of putative substrates. Oocytes expressing MasIne respond to hyperosmotic stimulation by releasing intracellular Ca(2+), as revealed by activation of the endogenous Ca(2+)-activated Cl(-) current. This Ca(2+) release requires the N-terminal 108 amino acid residues of MasIne and occurs via the inositol trisphosphate pathway. Fusion of the N terminus to the rat gamma-aminobutyric acid transporter (rGAT1) also renders rGAT1 responsive to hyperosmotic stimulation. Immunohistochemical analyses show that MasIne and Drosophila Ine have similar tissue distribution patterns, suggesting functional identity. Inebriated is expressed in tissues and cells actively involved in K(+) transport, which suggests that it may have a role in ion transport, particularly of K(+). We propose that stimulation of MasIne releases intracellular Ca(2+) in native tissues, activating Ca(2+)-dependent K(+) channels, and leading to K(+) transport. PMID- 11060216 TI - Hindlimb extensor muscle function during jumping and swimming in the toad (Bufo marinus). AB - Many anurans use their hindlimbs to generate propulsive forces during both jumping and swimming. To investigate the musculoskeletal dynamics and motor output underlying locomotion in such physically different environments, we examined patterns of muscle strain and activity using sonomicrometry and electromyography, respectively, during jumping and swimming in the toad Bufo marinus. We measured strain and electromyographic (EMG) activity in four hindlimb muscles: the semimembranosus, a hip extensor; the plantaris, an ankle extensor; and the gluteus and cruralis, two knee extensors. During jumping, these four muscles are activated approximately simultaneously; however, joint extension appears to be temporally staggered, with the hip beginning to extend prior to or initially faster than the more distal knee and ankle joints. Mirroring this pattern, the gluteus and plantaris shorten quite slowly and over a small distance during the first half of limb extension during take-off, before beginning to shorten rapidly. The hip and knee extensors finish shortening near the point of take-off (when the feet leave the ground), while the ankle-extending plantaris, which exhibits the longest-duration EMG burst, on average, always completes its shortening after take-off (mean 26 ms). During swimming, activation of the four muscles is also nearly synchronous at the start of a propulsive stroke. The onset of fascicle shortening is temporally staggered, with the knee extensors beginning to shorten first, prior to the hip and ankle extensors. In addition, the knee extensors also often exhibit some degree of slow passive shortening prior to the onset of EMG activity. The offset of muscle shortening during swimming is also staggered, and to a much greater extent than during jumping. During swimming, the cruralis and gluteus finish shortening first, the semimembranosus finishes 30-60 ms later, and the plantaris, which again exhibits the longest EMG burst, finishes shortening last (mean 150 ms after the cruralis). Interestingly, much of this extended shortening in the plantaris occurs at a relatively slow velocity and may reflect passive ankle extension caused by fluid forces, associated with previously generated unsteady (accelerative) limb movements, acting on the foot. Average EMG burst intensity tends to be greater during jumping than during swimming in all muscles but the gluteus. However, EMG burst duration only changes between jumping and swimming in the cruralis (duration during jumping is nearly twice as long as during swimming). The cruralis is also the only muscle to exhibit substantially greater fractional shortening during jumping (mean 0.28) than during swimming (mean 0.20 active strain, 0.22 total strain). On the basis of these results, it appears that toad hindlimb function is altered between jumping and swimming. Moreover, these functional differences are influenced by passive effects associated with physical differences between the external environments, but are also actively mediated by shifts in the motor output and mechanical behavior of several muscles. PMID- 11060217 TI - Mass spectrometric survey of peptides in cephalopods with an emphasis on the FMRFamide-related peptides. AB - A matrix-assisted laser desorption/ionization (MALDI) mass spectrometric (MS) survey of the major peptides in the stellar, fin and pallial nerves and the posterior chromatophore lobe of the cephalopods Sepia officinalis, Loligo opalescens and Dosidicus gigas has been performed. Although a large number of putative peptides are distinct among the three species, several molecular masses are conserved. In addition to peptides, characterization of the lipid content of the nerves is reported, and these lipid peaks account for many of the lower molecular masses observed. One conserved set of peaks corresponds to the FMRFamide-related peptides (FRPs). The Loligo opalescens FMRFa gene has been sequenced. It encodes a 331 amino acid residue prohormone that is processed into 14 FRPs, which are both predicted by the nucleotide sequence and confirmed by MALDI MS. The FRPs predicted by this gene (FMRFa, FLRFa/FIRFa and ALSGDAFLRFa) are observed in all three species, indicating that members of this peptide family are highly conserved across cephalopods. PMID- 11060218 TI - Novel aspects of the transport of organic anions by the malpighian tubules of Drosophila melanogaster. AB - Para-aminohippuric acid (PAH) is a negatively charged organic ion that can pass across the epithelium of Malpighian tubules. Its mode of transport was studied in Malpighian tubules of Drosophila melanogaster. PAH transport was an active process, with a K(m) of 2. 74 mmol l(-)(1) and a V(max) of 88.8 pmol min(-)(1). Tubules had a low passive permeability to PAH, but PAH transport rates (832 nmol min(-)(1 )mm(2)) and concentrative ability ([PAH](secreted fluid):[PAH](bath)=81.2) were the highest measured to date for insects. Competition experiments indicated that there were two organic anion transporters, one that transports carboxylate compounds, such as PAH and fluorescein, and another that transports sulphonates, such as amaranth and Indigo Carmine. PAH transport appears to be maximal in vivo because the rate of transport by isolated tubules is not increased when these are challenged with cyclic AMP, cyclic GMP, leucokinin I or staurosporine. Basolateral PAH transport was inhibited by ouabain and dependent on the Na(+) gradient. The Malpighian tubules appeared not to possess an organic acid/ &agr; -keto acid exchanger because PAH accumulation was not affected by low concentrations (100 micromol l(-)(1)) of &agr; -keto acids ( &agr; -ketoglutarate, glutarate, citrate and succinate) or the activity of phosphokinase C. PAH transport may be directly coupled to the Na(+) gradient, perhaps via Na(+)/organic acid cotransport. Fluorescence microscopy showed that transport of the carboxylate fluorescein was confined to the principal cells of the main (secretory) segment and all the cells of the lower (reabsorptive) segment. Organic anions were transported across the cytoplasm of the principal cells both by diffusion and in vesicles. The accumulation of punctate fluorescence in the lumen is consistent with exocytosis of the cytoplasmic vesicles. Apical PAH transport was independent of the apical membrane potential and may not occur by an electrodiffusive mechanism. PMID- 11060219 TI - Function of the heterocercal tail in white sturgeon: flow visualization during steady swimming and vertical maneuvering. AB - Basal ray-finned fishes possess a heterocercal tail in which the dorsal lobe containing the extension of the vertebral column is longer than the ventral lobe. Clarifying the function of the heterocercal tail has proved elusive because of the difficulty of measuring the direction of force produced relative to body position in the aquatic medium. We measured the direction of force produced by the heterocercal tail of the white sturgeon (Acipenser transmontanus) by visualizing flow in the wake of the tail using digital particle image velocimetry (DPIV) while simultaneously recording body position and motion using high-speed video. To quantify tail function, we measured the vertical body velocity, the body angle and the path angle of the body from video recordings and the vortex ring axis angle and vortex jet angle from DPIV recordings of the wake downstream from the tail. These variables were measured for sturgeon exhibiting three swimming behaviors at 1.2 L s(-)(1), where L is total body length: rising through the water column, holding vertical position, and sinking through the water column. For vertical body velocity, body angle and path angle values, all behaviors were significantly different from one another. For vortex ring axis angle and vortex jet angle, rising and holding behavior were not significantly different from each other, but both were significantly different from sinking behavior. During steady horizontal swimming, the sturgeon tail generates a lift force relative to the path of motion but no rotational moment because the reaction force passes through the center of mass. For a rising sturgeon, the tail does not produce a lift force but causes the tail to rotate ventrally in relation to the head since the reaction force passes ventral to the center of mass. While sinking, the direction of the fluid jet produced by the tail relative to the path of motion causes a lift force to be created and causes the tail to rotate dorsally in relation to the head since the reaction force passes dorsal to the center of mass. These data provide evidence that sturgeon can actively control the direction of force produced by their tail while maneuvering through the water column because the relationship between vortex jet angle and body angle is not constant. PMID- 11060220 TI - Modulatory effects of nitric oxide on synaptic depression in the crayfish neuromuscular system. AB - A characteristic physiological property of the neuromuscular junction between giant motor neurones (MoGs) and fast flexor muscles in crayfish is synaptic depression, in which repetitive electrical stimulation of the MoG results in a progressive decrease in excitatory junction potential (EJP) amplitude in flexor muscle fibres. Previous studies have demonstrated that l-arginine (l-Arg) modulates neuromuscular transmission. Since l-Arg is a precursor of nitric oxide (NO), we examined the possibility that NO may be involved in modulating neuromuscular transmission from MoGs to abdominal fast flexor muscles. The effect of a NO-generating compound, NOC7, was similar to that of l-Arg, reversibly decreasing the EJP amplitude mediated by the MoG. While NOC7 reduced the amplitude of the EJP, it induced no significant change in synaptic depression. In contrast, a scavenger of free radical NO, carboxy-PTIO, and an inhibitor of nitric oxide synthase, l-NAME, reversibly increased the EJP amplitude mediated by MoGs. Synaptic depression mediated by repetitive stimulation of MoGs at 1 Hz was partially blocked by bath application of l-NAME. Bath application of a NO scavenger, a NOS inhibitor and NO-generating compounds had no significant effects on the depolarisation of the muscle fibres evoked by local application of l glutamate. The opposing effects on EJP amplitude of NOC7 and of carboxy-PTIO and l-NAME suggest that endogenous NO presynaptically modulates neuromuscular transmission and that it could play a prominent role at nerve terminals in eliciting MoG-mediated synaptic depression in the crayfish Procambarus clarkii. PMID- 11060221 TI - Processes contributing to metabolic depression in hepatopancreas cells from the snail Helix aspersa. AB - Cells isolated from the hepatopancreas of the land snail Helix aspersa strongly depress respiration both immediately in response to lowered P(O2) (oxygen conformation) and, in the longer term, during aestivation. These phenomena were analysed by dividing cellular respiration into non-mitochondrial and mitochondrial respiration using the mitochondrial poisons myxothiazol, antimycin and azide. Non-mitochondrial respiration accounted for a surprisingly large proportion, 65+/-5 %, of cellular respiration in control cells at 70 % air saturation. Non-mitochondrial respiration decreased substantially as oxygen tension was lowered, but mitochondrial respiration did not, and the oxygen conforming behaviour of the cells was due entirely to the oxygen-dependence of non-mitochondrial oxygen consumption. Non-mitochondrial respiration was still responsible for 45+/-2 % of cellular respiration at physiological oxygen tension. Mitochondrial respiration was further subdivided into respiration used to drive ATP turnover and respiration used to drive futile proton cycling across the mitochondrial inner membrane using the ATP synthase inhibitor oligomycin. At physiological oxygen tensions, 34+/-5 % of cellular respiration was used to drive ATP turnover and 22+/-4 % was used to drive proton cycling, echoing the metabolic inefficiency previously observed in liver cells from mammals, reptiles and amphibians. The respiration rate of hepatopancreas cells from aestivating snails was only 37 % of the control value. This was caused by proportional decreases in non-mitochondrial and mitochondrial respiration and in respiration to drive ATP turnover and to drive proton cycling. Thus, the fraction of cellular respiration devoted to different processes remained constant and the cellular energy balance was preserved in the hypometabolic state. PMID- 11060222 TI - Catalytic activity and heat production by the Ca(2+)-ATPase from sea cucumber (Ludwigothurea grisea) longitudinal smooth muscle: modulation by monovalent cations. AB - In muscle cells, excitation-contraction coupling involves the translocation of Ca(2+) between intracellular compartments and the cytosol. Heat derived from the hydrolysis of ATP by the sarcoplasmic reticulum Ca(2+)-ATPase of skeletal muscle plays an important role in the thermoregulation and energy balance of the cell. Although several Ca(2+)-ATPase isoforms have been described in vertebrates, little is known about Ca(2+) transport in invertebrates. In this report, a Ca(2+) ATPase is identified in the microsomal fraction obtained from sea cucumber (Ludwigothurea grisea) smooth muscle. The activity of this enzyme is enhanced three- to fivefold by K(+) and Na(+). During Ca(2+) transport, the ATPase can synthesise ATP from ADP and inorganic phosphate (P(i)) using the energy derived from the Ca(2+) gradient formed across the microsomal membrane (ATP<->P(i) exchange). The apparent affinity of the enzyme for P(i) is increased by more than one order of magnitude by K(+). In the presence of K(+), the fraction of ATP synthesised during the exchange reaction by sea cucumber microsomes was found to be larger than that measured in microsomes derived from either rabbit or trout muscle. Like the isoforms found in skeletal muscle, the sea cucumber Ca(2+) ATPase can convert osmotic energy into heat. The amount of heat produced after the hydrolysis of each ATP molecule increases two- to threefold when a Ca(2+) gradient is formed across the microsomal membrane. PMID- 11060223 TI - Hearing threshold and frequency discrimination in the purely aquatic frog Xenopus laevis (Pipidae): measurement by means of conditioning. AB - Hearing threshold and frequency discrimination for underwater sound were measured in the clawed frog Xenopus laevis by means of conditioning. A go/no go discrimination procedure was used in which the test tone was presented concurrently with a wave on the surface of the water. The tone signalled whether or not the frog should respond to the wave. The hearing range of X. laevis was 200-4000 Hz. Similar thresholds of 92-96 dB re 1 microPa were found at 600 Hz, 1400-1800 Hz and 3200-3600 Hz. A high threshold at 1000-1300 Hz suggested that this was the frequency range between the sensitivities of the amphibian and basilar papillae. Relative frequency discrimination was approximately 5 % at 400 800 Hz, 45 % at 1000 Hz and 2.4-6 % at 1600-2500 Hz. This last range encompasses the dominant frequencies of the advertisement call of this species. High discrimination acuity at these frequencies may be used in distinguishing between calling males. The threshold for a one-third-octave bandpass noise centred at 600 Hz was 27.6 dB lower than that for a pure tone of 600 Hz, suggesting that sound intensity was integrated within this bandwidth, possibly by a critical-band mechanism. PMID- 11060224 TI - Thyroid hormone concentrations in captive and free-ranging West Indian manatees (Trichechus manatus). AB - Because thyroid hormones play a critical role in the regulation of metabolism, the low metabolic rates reported for manatees suggest that thyroid hormone concentrations in these animals may also be reduced. However, thyroid hormone concentrations have yet to be examined in manatees. The effects of captivity, diet and water salinity on plasma total triiodothyronine (tT(3)), total thyroxine (tT(4)) and free thyroxine (fT(4)) concentrations were assessed in adult West Indian manatees (Trichechus manatus). Free-ranging manatees exhibited significantly greater tT(4) and fT(4) concentrations than captive adults, regardless of diet, indicating that some aspect of a captive existence results in reduced T(4) concentrations. To determine whether this reduction might be related to feeding, captive adults fed on a mixed vegetable diet were switched to a strictly sea grass diet, resulting in decreased food consumption and a decrease in body mass. However, tT(4) and fT(4) concentrations were significantly elevated over initial values for 19 days. This may indicate that during periods of reduced food consumption manatees activate thyroid-hormone-promoted lipolysis to meet water and energetic requirements. Alterations in water salinity for captive animals did not induce significant changes in thyroid hormone concentrations. In spite of lower metabolic rates, thyroid hormone concentrations in captive manatees were comparable with those for other terrestrial and marine mammals, suggesting that the low metabolic rate in manatees is not attributable to reduced circulating thyroid hormone concentrations. PMID- 11060225 TI - Sexual dimorphism in forelimb muscles of the bullfrog, Rana catesbeiana: a functional analysis of isometric contractile properties. AB - In many species of frog, the forelimb muscles important in amplexus are known to be much larger in males than in females. We studied this dimorphism in three forelimb muscles in the bullfrog [abductor indicus longus (AIL), flexor carpi radialis (FCR) and extensor carpi radialis (ECR)] by testing their isometric contractile properties. One muscle that is not dimorphic, the extensor carpi ulnaris (ECU), was also studied as a control. In addition to being greater in wet mass and in cross-sectional area in the males, our data show that the dimorphic muscles also produce significantly larger isometric forces in males than in females. The tetanic force per cm(2) of muscle cross-sectional area did not differ between the sexes, so that force within a muscle varies directly with muscle size. However, a number of the contractile variables we measured show that male muscles differ functionally from those of females. The male twitch contraction times were significantly longer in the AIL, and the male half relaxation times were longer in both the AIL and FCR. These two dimorphic muscles were also significantly less fatiguable in males than were the corresponding female muscles. Their higher endurance resulted from the maintenance of high levels of unrelaxed force sustained between trains of stimuli during the fatigue test. This sustained force is much less pronounced in the female muscles, suggesting that high levels of sustained force may be a key functional feature that enables males to maintain amplexus economically for prolonged periods. PMID- 11060226 TI - Foraging energetics and diving behavior of lactating New Zealand sea lions, Phocarctos hookeri. AB - The New Zealand sea lion, Phocarctos hookeri, is the deepest- and longest-diving sea lion. We were interested in whether the diving ability of this animal was related to changes in its at-sea and diving metabolic rates. We measured the metabolic rate, water turnover and diving behavior of 12 lactating New Zealand sea lions at Sandy Bay, Enderby Island, Auckland Islands Group, New Zealand (50 degrees 30'S, 166 degrees 17'E), during January and February 1997 when their pups were between 1 and 2 months old. Metabolic rate (rate of CO(2) production) and water turnover were measured using the (18)O doubly-labeled water technique, and diving behavior was measured with time/depth recorders (TDRs). Mean total body water was 66.0+/-1.1 % (mean +/- s.d.) and mean rate of CO(2) production was 0. 835+/-0.114 ml g(-)(1 )h(-)(1), which provides an estimated mass-specific field metabolic rate (FMR) of 5.47+/-0.75 W kg(-)(1). After correction for time on shore, the at-sea FMR was estimated to be 6.65+/-1.09 W kg(-)(1), a value 5.8 times the predicted standard metabolic rate of a terrestrial animal of equal size. The mean maximum dive depth was 353+/-164 m, with a mean diving depth of 124+/-36 m. The mean maximum dive duration was 8.3+/-1.7 min, with an average duration of 3.4+/-0.6 min. The deepest, 550 m, and longest, 11.5 min, dives were made by the largest animal (155 kg). Our results indicate that the deep and long duration diving ability of New Zealand sea lions is not due to a decreased diving metabolic rate. Individual sea lions that performed deeper dives had lower FMRs, which may result from the use of energetically efficient burst-and-glide locomotion. There are differences in the foraging patterns of deep and shallow divers that may reflect differences in surface swimming, time spent on the surface and/or diet. Our data indicate that, although New Zealand sea lions have increased their O(2) storage capacity, they do not, or cannot, significantly reduce their at-sea metabolic rates and are therefore likely to be operating near their physiological maximum. PMID- 11060227 TI - Relative metabolic utilization of in situ rabbit soleus muscle: thermistor-based measurements and model. AB - Electrically conditioned skeletal muscle can provide the continuous power source for cardiac assistance devices. Optimization of the available sustained power from in vivo skeletal muscle requires knowledge of its metabolic utilization and constraints. A thermistor-based technique has been developed to measure temperature changes and to provide a relative estimate for metabolic utilization of in situ rabbit soleus muscle. The relative thermistor response, active tension and muscle displacement were measured during cyclic isometric and isotonic contractions across a range of muscle tensions and contraction durations. The thermistor response demonstrated linear relationships versus both contraction duration at a fixed muscle length and active tension at a fixed contraction duration (r(2)=0.90+/-0.14 and 0.70+/-0.21, respectively; means +/- s.d.). A multiple linear regression model was developed to predict normalized thermistor response, DeltaT, across a range of conditions. Significant model variables were identified using a backward stepwise regression procedure. The relationships for the in situ muscles were qualitatively similar to those reported for mammalian in vitro muscle fiber preparations. The model had the form DeltaT=C+at(c)F+bW, where the constant C, and coefficients for the contraction duration t(c) (ms), normalized active tension F and normalized net work W were C=-1.00 (P<0.001), a=5.97 (P<0.001) and b=2.12 (P<0.001). PMID- 11060228 TI - The Gsh2 homeodomain gene controls multiple aspects of telencephalic development. AB - Homeobox genes have recently been demonstrated to be important for the proper patterning of the mammalian telencephalon. One of these genes is Gsh2, whose expression in the forebrain is restricted to the ventral domain. In this study, we demonstrate that Gsh2 is a downstream target of sonic hedgehog and that lack of Gsh2 results in profound defects in telencephalic development. Gsh2 mutants have a significant decrease in the expression of numerous genes that mark early development of the lateral ganglionic eminence, the striatal anlage. Accompanying this early loss of patterning genes is an initial expansion of dorsal telencephalic markers across the cortical-striatal boundary into the lateral ganglionic eminence. Interestingly, as development proceeds, there is compensation for this early loss of markers that is coincident with a molecular re-establishment of the cortical-striatal boundary. Despite this compensation, there is a defect in the development of distinct subpopulations of striatal neurons. Moreover, while our analysis suggests that the migration of the ventrally derived interneurons to the developing cerebral cortex is not significantly affected in Gsh2 mutants, there is a distinct delay in the appearance of GABAergic interneurons in the olfactory bulb. Taken together, our data support a model in which Gsh2, in response to sonic hedgehog signaling, plays a crucial role in multiple aspects of telencephalic development. PMID- 11060229 TI - Meiotic maturation induces animal-vegetal asymmetric distribution of aPKC and ASIP/PAR-3 in Xenopus oocytes. AB - The asymmetric distribution of cellular components is an important clue for understanding cell fate decision during embryonic patterning and cell functioning after differentiation. In C. elegans embryos, PAR-3 and aPKC form a complex that colocalizes to the anterior periphery of the one-cell embryo, and are indispensable for anterior-posterior polarity that is formed prior to asymmetric cell division. In mammals, ASIP (PAR-3 homologue) and aPKCgamma form a complex and colocalize to the epithelial tight junctions, which play critical roles in epithelial cell polarity. Although the mechanism by which PAR-3/ASIP and aPKC regulate cell polarization remains to be clarified, evolutionary conservation of the PAR-3/ASIP-aPKC complex suggests their general role in cell polarity organization. Here, we show the presence of the protein complex in Xenopus laevis. In epithelial cells, XASIP and XaPKC colocalize to the cell-cell contact region. To our surprise, they also colocalize to the animal hemisphere of mature oocytes, whereas they localize uniformly in immature oocytes. Moreover, hormonal stimulation of immature oocytes results in a change in the distribution of XaPKC 2-3 hours after the completion of germinal vesicle breakdown, which requires the kinase activity of aPKC. These results suggest that meiotic maturation induces the animal-vegetal asymmetry of aPKC. PMID- 11060230 TI - Two homeobox genes define the domain of EphA3 expression in the developing chick retina. AB - Graded expression of the Eph receptor EphA3 in the retina and its two ligands, ephrin A2 and ephrin A5 in the optic tectum, the primary target of retinal axons, have been implicated in the formation of the retinotectal projection map. Two homeobox containing genes, SOHo1 and GH6, are expressed in a nasal-high, temporal low pattern during early retinal development, and thus in opposing gradients to EphA3. Retroviral misexpression of SOHo1 or GH6 completely and specifically repressed EphA3 expression in the neural retina, but not in other parts of the central nervous system, such as the optic tectum. Under these conditions, some temporal ganglion cell axons overshot their expected termination zones in the rostral optic tectum, terminating aberrantly at more posterior locations. However, the majority of ganglion cell axons mapped to the appropriate rostrocaudal locations, although they formed somewhat more diffuse termination zones. These findings indicate that other mechanisms, in addition to differential EphA3 expression in the neural retina, are required for retinal ganglion axons to map to the appropriate rostrocaudal locations in the optic tectum. They further suggest that the control of topographic specificity along the retinal nasal temporal axis is split into several independent pathways already at a very early time in development. PMID- 11060231 TI - Patterning of the C. elegans 1 degrees vulval lineage by RAS and Wnt pathways. AB - In C. elegans, the descendants of the 1 degrees vulval precursor cell (VPC) establish a fixed spatial pattern of two different cell fates: E-F-F-E. The two inner granddaughters attach to the somatic gonadal anchor cell (AC) and generate four vulF cells, while the two outer granddaughters produce four vulE progeny. zmp-1::GFP, a molecular marker that distinguishes these two fates, is expressed in vulE cells, but not vulF cells. We demonstrate that a short-range AC signal is required to ensure that the pattern of vulE and vulF fates is properly established. In addition, signaling between the inner and outer 1 degrees VPC descendants, as well as intrinsic polarity of the 1 degrees VPC daughters, is involved in the asymmetric divisions of the 1 degrees VPC daughters and the proper orientation of the outcome. Finally, we provide evidence that RAS signaling is used during this new AC signaling event, while the Wnt receptor LIN 17 appears to mediate signaling between the inner and outer 1 degrees VPC descendants. PMID- 11060232 TI - Early myotome specification regulates PDGFA expression and axial skeleton development. AB - Reciprocal defects in signaling between the myotome and the sclerotome compartments of the somites in PDGFRalpha and Myf5 mutant embryos lead to alterations in the formation of the vertebrae and the ribs. To investigate the significance of these observations, we have examined the role of PDGF signaling in the developing somite. PDGFA ligand expression was not detected in the myotome of Myf5 null mutant embryos and PDGFA promoter activity was regulated by Myf5 in vitro. PDGFA stimulated chondrogenesis in somite micromass cultures as well as in embryos when PDGFA was knocked into the Myf5 locus, resulting in increased vertebral and rib development. PDGFA expression in the myotome was fully restored in embryos in which MyoD has been introduced at the Myf5 locus but to a lesser extent in similar myogenin knock-in embryos. These results underscore the importance of growth factor signaling within the developing somite and suggest an important role for myogenic determination factors in orchestrating normal development of the axial skeleton. PMID- 11060233 TI - The C. elegans F-box/WD-repeat protein LIN-23 functions to limit cell division during development. AB - In multicellular eukaryotes, a complex program of developmental signals regulates cell growth and division by controlling the synthesis, activation and degradation of G(1) cell cycle regulators. Here we describe the lin-23 gene of Caenorhabditis elegans, which is required to restrain cell proliferation in response to developmental cues. In lin-23 null mutants, all postembryonic blast cells undergo extra divisions, creating supernumerary cells that can differentiate and function normally. In contrast to the inability to regulate the extent of blast cell division in lin-23 mutants, the timing of initial cell cycle entry of blast cells is not affected. lin-23 encodes an F-box/WD-repeat protein that is orthologous to the Saccharomyces cerevisiae gene MET30, the Drosophila melanogaster gene slmb and the human gene betaTRCP, all of which function as components of SCF ubiquitin ligase complexes. Loss of function of the Drosophila slmb gene causes the growth of ectopic appendages in a non-cell autonomous manner. In contrast, lin-23 functions cell autonomously to negatively regulate cell cycle progression, thereby allowing cell cycle exit in response to developmental signals. PMID- 11060234 TI - Temporally restricted expression of transcription factor betaFTZ-F1: significance for embryogenesis, molting and metamorphosis in Drosophila melanogaster. AB - FTZ-F1, a member of the nuclear receptor superfamily, has been implicated in the activation of the segmentation gene fushi tarazu during early embryogenesis of Drosophila melanogaster. We found that an isoform of FTZ-F1, betaFTZ-F1, is expressed in the nuclei of almost all tissues slightly before the first and second larval ecdysis and before pupation. Severely affected ftz-f1 mutants display an embryonic lethal phenotype, but can be rescued by ectopic expression of betaFTZ-F1 during the period of endogenous betaFTZ-F1 expression in the wild type. The resulting larvae are not able to molt, but this activity is rescued again by forced expression of betaFTZ-F1, allowing progression to the next larval instar stage. On the other hand, premature expression of betaFTZ-F1 in wild-type larvae at mid-first instar or mid-second instar stages causes defects in the molting process. Sensitive periods were found to be around the time of peak ecdysteroid levels and slightly before the start of endogenous betaFTZ-F1 expression. A hypomorphic ftz-f1 mutant that arrests in the prepupal stage can also be rescued by ectopic, time-specific expression of betaFTZ-F1. Failure of salivary gland histolysis, one of the phenotypes of the ftz-f1 mutant, is rescued by forced expression of the ftz-f1 downstream gene BR-C during the late prepupal period. These results suggest that betaFTZ-F1 regulates genes associated with ecdysis and metamorphosis, and that the exact timing of its action in the ecdysone-induced gene cascade is important for proper development. PMID- 11060235 TI - Loss- and gain-of-function mutations show a polycomb group function for Ring1A in mice. AB - The products of the Polycomb group (PcG) of genes act as transcriptional repressors involved in the maintenance of homeotic gene expression patterns throughout development, from flies to mice. Biochemical and molecular evidence suggests that the mouse Ring1A gene is a member of the PcG of genes. However, genetic evidence is needed to establish PcG function for Ring1A, since contrary to all other murine PcG genes, there is no known Drosophila PcG gene encoding a homolog of the Ring1A protein. To study Ring1A function we have generated a mouse line lacking Ring1A and mouse lines overexpressing Ring1A. Both Ring1A(-/-)and Ring1A(+/-) mice show anterior transformations and other abnormalities of the axial skeleton, which indicates an unusual sensitivity of axial skeleton patterning to Ring1A gene dosage. Ectopic expression of Ring1A also results in dose-dependent anterior transformations of vertebral identity, many of which, interestingly, are shared by Ring1A(-/-) mice. In contrast, the alterations of Hox gene expression observed in both type of mutant mice are subtle and involve a reduced number of Hox genes. Taken together, these results provide genetic evidence for a PcG function of the mouse Ring1A gene. PMID- 11060236 TI - Asymmetric nodal signaling in the zebrafish diencephalon positions the pineal organ. AB - The vertebrate brain develops from a bilaterally symmetric neural tube but later displays profound anatomical and functional asymmetries. Despite considerable progress in deciphering mechanisms of visceral organ laterality, the genetic pathways regulating brain asymmetries are unknown. In zebrafish, genes implicated in laterality of the viscera (cyclops/nodal, antivin/lefty and pitx2) are coexpressed on the left side of the embryonic dorsal diencephalon, within a region corresponding to the presumptive epiphysis or pineal organ. Asymmetric gene expression in the brain requires an intact midline and Nodal-related factors. RNA-mediated rescue of mutants defective in Nodal signaling corrects tissue patterning at gastrulation, but fails to restore left-sided gene expression in the diencephalon. Such embryos develop into viable adults with seemingly normal brain morphology. However, the pineal organ, which typically emanates at a left-to-medial site from the dorsal diencephalic roof, becomes displaced in position. Thus, a conserved signaling pathway regulating visceral laterality also underlies an anatomical asymmetry of the zebrafish forebrain. PMID- 11060237 TI - A micromere induction signal is activated by beta-catenin and acts through notch to initiate specification of secondary mesenchyme cells in the sea urchin embryo. AB - At fourth cleavage of sea urchin embryos four micromeres at the vegetal pole separate from four macromeres just above them in an unequal cleavage. The micromeres have the capacity to induce a second axis if transplanted to the animal pole and the absence of micromeres at the vegetal pole results in the failure of macromere progeny to specify secondary mesenchyme cells (SMCs). This suggests that micromeres have the capacity to induce SMCs. We demonstrate that micromeres require nuclear beta-catenin to exhibit SMC induction activity. Transplantation studies show that much of the vegetal hemisphere is competent to receive the induction signal. The micromeres induce SMCs, most likely through direct contact with macromere progeny, or at most a cell diameter away. The induction is quantitative in that more SMCs are induced by four micromeres than by one. Temporal studies show that the induction signal is passed from the micromeres to macromere progeny between the eighth and tenth cleavage. If micromeres are removed from hosts at the fourth cleavage, SMC induction in hosts is rescued if they later receive transplanted micromeres between the eighth and tenth cleavage. After the tenth cleavage addition of induction-competent micromeres to micromereless embryos fails to specify SMCs. For macromere progeny to be competent to receive the micromere induction signal, beta-catenin must enter macromere nuclei. The macromere progeny receive the micromere induction signal through the Notch receptor. Signaling-competent micromeres fail to induce SMCs if macromeres express dominant-negative Notch. Expression of an activated Notch construct in macromeres rescues SMC specification in the absence of induction-competent micromeres. These data are consistent with a model whereby beta-catenin enters the nuclei of micromeres and, as a consequence, the micromeres produce an inductive ligand. Between the eighth and tenth cleavage micromeres induce SMCs through Notch. In order to be receptive to the micromere inductive signal the macromeres first must transport beta-catenin to their nuclei, and as one consequence the Notch pathway becomes competent to receive the micromere induction signal, and to transduce that signal. As Notch is maternally expressed in macromeres, additional components must be downstream of nuclear beta catenin in macromeres for these cells to receive and transduce the micromere induction signal. PMID- 11060238 TI - Hereditary spherocytosis in zebrafish riesling illustrates evolution of erythroid beta-spectrin structure, and function in red cell morphogenesis and membrane stability. AB - Spectrins are key cytoskeleton proteins with roles in membrane integrity, cell morphology, organelle transport and cell polarity of varied cell types during development. Defects in erythroid spectrins in humans result in congenital hemolytic anemias with altered red cell morphology. Although well characterized in mammals and invertebrates, analysis of the structure and function of non mammalian vertebrate spectrins has been lacking. The zebrafish riesling (ris) suffers from profound anemia, where the developing red cells fail to assume terminally differentiated erythroid morphology. Using comparative genomics, erythroid beta-spectrin (sptb) was identified as the gene mutated in ris. Zebrafish Sptb shares 62.3% overall identity with the human ortholog and phylogenetic comparisons suggest intragenic duplication and divergence during evolution. Unlike the human and murine orthologs, the pleckstrin homology domain of zebrafish Sptb is not removed in red cells by alternative splicing. In addition, apoptosis and abnormal microtubule marginal band aggregation contribute to hemolysis of mutant erythrocytes, which are features not present in mammalian red cells with sptb defects. This study presents the first genetic characterization of a non-mammalian vertebrate sptb and demonstrates novel features of red cell hemolysis in non-mammalian red cells. Further, we propose that the distinct mammalian erythroid morphology may have evolved from specific modifications of Sptb structure and function. PMID- 11060239 TI - Evidence that members of the Cut/Cux/CDP family may be involved in AER positioning and polarizing activity during chick limb development. AB - In vertebrates, the apical ectodermal ridge (AER) is a specialized epithelium localized at the dorsoventral boundary of the limb bud that regulates limb outgrowth. In Drosophila, the wing margin is also a specialized region located at the dorsoventral frontier of the wing imaginal disc. The wingless and Notch pathways have been implicated in positioning both the wing margin and the AER. One of the nuclear effectors of the Notch signal in the wing margin is the transcription factor cut. Here we report the identification of two chick homologues of the Cut/Cux/CDP family that are expressed in the developing limb bud. Chick cux1 is expressed in the ectoderm outside the AER, as well as around ridge-like structures induced by (&bgr;)-catenin, a downstream target of the Wnt pathway. cux1 overexpression in the chick limb results in scalloping of the AER and limb truncations, suggesting that Cux1 may have a role in limiting the position of the AER by preventing the ectodermal cells around it from differentiating into AER cells. The second molecule of the Cut family identified in this study, cux2, is expressed in the pre-limb lateral plate mesoderm, posterior limb bud and flank mesenchyme, a pattern reminiscent of the distribution of polarizing activity. The polarizing activity is determined by the ability of a certain region to induce digit duplications when grafted into the anterior margin of a host limb bud. Several manipulations of the chick limb bud show that cux2 expression is regulated by retinoic acid, Sonic hedgehog and the posterior AER. These results suggest that Cux2 may have a role in generating or mediating polarizing activity. Taking into account the probable involvement of Cut/Cux/CDP molecules in cell cycle regulation and differentiation, our results raise the hypothesis that chick Cux1 and Cux2 may act by modulating proliferation versus differentiation in the limb ectoderm and polarizing activity regions, respectively. PMID- 11060240 TI - The maternal NF-kappaB/dorsal gradient of Tribolium castaneum: dynamics of early dorsoventral patterning in a short-germ beetle. AB - In the long-germ insect Drosophila melanogaster dorsoventral polarity is induced by localized Toll-receptor activation which leads to the formation of a nuclear gradient of the rel/ NF-kappaB protein Dorsal. Peak levels of nuclear Dorsal are found in a ventral stripe spanning the entire length of the blastoderm embryo allowing all segments and their dorsoventral subdivisions to be synchronously specified before gastrulation. We show that a nuclear Dorsal protein gradient of similar anteroposterior extension exists in the short-germ beetle, Tribolium castaneum, which forms most segments from a posterior growth zone after gastrulation. In contrast to Drosophila, (i) nuclear accumulation is first uniform and then becomes progressively restricted to a narrow ventral stripe, (ii) gradient refinement is accompanied by changes in the zygotic expression of the Tribolium Toll-receptor suggesting feedback regulation and, (iii) the gradient only transiently overlaps with the expression of a potential target, the Tribolium twist homolog, and does not repress Tribolium decapentaplegic. No nuclear Dorsal is seen in the cells of the growth zone of Tribolium embryos, indicating that here dorsoventral patterning occurs by a different mechanism. However, Dorsal is up-regulated and transiently forms a nuclear gradient in the serosa, a protective extraembryonic cell layer ultimately covering the whole embryo. PMID- 11060241 TI - PIN-FORMED 1 regulates cell fate at the periphery of the shoot apical meristem. AB - The process of organ positioning has been addressed, using the pin-formed 1 (pin1) mutant as a tool. PIN1 is a transmembrane protein involved in auxin transport in Arabidopsis. Loss of function severely affects organ initiation, and pin1 mutants are characterised by an inflorescence meristem that does not initiate any flowers, resulting in the formation of a naked inflorescence stem. This phenotype, combined with the proposed role of PIN1 in hormone transport, makes the mutant an ideal tool to study organ formation and phyllotaxis, and here we present a detailed analysis of the molecular modifications at the shoot apex caused by the mutation. We show that meristem structure and function are not severely affected in the mutant. Major alterations, however, are observed at the periphery of the pin1 meristem, where organ initiation should occur. Although two very early markers of organ initiation, LEAFY and AINTEGUMENTA, are expressed at the periphery of the mutant meristem, the cells are not recruited into distinct primordia. Instead a ring-like domain expressing those primordium specific genes is observed around the meristem. This ring-like domain also expresses a boundary marker, CUP-SHAPED COTYLEDON 2, involved in organ separation, showing that the zone at the meristem periphery has a hybrid identity. This implies that PIN1 is not only involved in organ outgrowth, but that it is also necessary for organ separation and positioning. A model is presented in which PIN1 and the local distribution of auxin control phyllotaxis. PMID- 11060242 TI - A role for Pax6 in the normal development of dorsal thalamus and its cortical connections. AB - The transcription factor Pax6 is widely expressed throughout the developing nervous system, including most alar regions of the newly formed murine diencephalon. Later in embryogenesis its diencephalic expression becomes more restricted. It persists in the developing anterior thalamus (conventionally termed "ventral" thalamus) and pretectum but is downregulated in the body of the posterior (dorsal) thalamus. At the time of this downregulation, the dorsal thalamus forms its major axonal efferent pathway via the ventral telencephalon to the cerebral cortex. This pathway is absent in mice lacking functional Pax6 (small eye homozygotes: Sey/Sey). We tested whether the mechanism underlying this defect includes abnormalities of the dorsal thalamus itself. We exploited a new transgenic mouse ubiquitously expressing green fluorescent protein tagged with tau, in which axonal tracts are clearly visible, and co-cultured dorsal thalamic explants from Pax6(+/+ )or Pax6(Sey/Sey )embryos carrying the transgene with wild type tissues from other regions of the forebrain. Whereas Pax6(+/+ )thalamic explants produced strong innervation of wild-type ventral telencephalic explants in a pattern that mimicked the thalamocortical tract in vivo, Pax6(Sey)(/Sey) explants did not, indicating a defect in the ability of mutant dorsal thalamic cells to respond to signals normally present in ventral telencephalon. Pax6(Sey)(/Sey) embryos also showed early alterations in the expression of regulatory genes in the region destined to become dorsal thalamus. Whereas in normal mice Nkx2.2 and Lim1/Lhx1 are expressed ventral to this region, in the mutants their expression domains are throughout it, suggesting that a primary action of Pax6 is to generate correct dorsoventral patterning in the diencephalon. Our results suggest that normal thalamocortical development requires the actions of Pax6 within the dorsal thalamus itself. PMID- 11060243 TI - Overlapping roles of two Hox genes and the exd ortholog ceh-20 in diversification of the C. elegans postembryonic mesoderm. AB - Members of the Hox family of homeoproteins and their cofactors play a central role in pattern formation of all germ layers. During postembryonic development of C. elegans, non-gonadal mesoderm arises from a single mesoblast cell M. Starting in the first larval stage, M divides to produce 14 striated muscles, 16 non striated muscles, and two non-muscle cells (coelomocytes). We investigated the role of the C. elegans Hox cluster and of the exd ortholog ceh-20 in patterning of the postembryonic mesoderm. By examining the M lineage and its differentiation products in different Hox mutant combinations, we found an essential but overlapping role for two of the Hox cluster genes, lin-39 and mab-5, in diversification of the postembryonic mesoderm. This role of the two Hox gene products required the CEH-20 cofactor. One target of these two Hox genes is the C. elegans twist ortholog hlh-8. Using both in vitro and in vivo assays, we demonstrated that twist is a direct target of Hox activation. We present evidence from mutant phenotypes that twist is not the only target for Hox genes in the M lineage: in particular we show that lin-39 mab-5 double mutants exhibit a more severe M lineage defect than the hlh-8 null mutant. PMID- 11060244 TI - The Phox2b transcription factor coordinately regulates neuronal cell cycle exit and identity. AB - In the vertebrate neural tube, cell cycle exit of neuronal progenitors is accompanied by the expression of transcription factors that define their generic and sub-type specific properties, but how the regulation of cell cycle withdrawal intersects with that of cell fate determination is poorly understood. Here we show by both loss- and gain-of-function experiments that the neuronal-subtype specific homeodomain transcription factor Phox2b drives progenitor cells to become post-mitotic. In the absence of Phox2b, post-mitotic neuronal precursors are not generated in proper numbers. Conversely, forced expression of Phox2b in the embryonic chick spinal cord drives ventricular zone progenitors to become post-mitotic neurons and to relocate to the mantle layer. In the neurons thus generated, ectopic expression of Phox2b is sufficient to initiate a programme of motor neuronal differentiation characterised by expression of Islet1 and of the cholinergic transmitter phenotype, in line with our previous results showing that Phox2b is an essential determinant of cranial motor neurons. These results suggest that Phox2b coordinates quantitative and qualitative aspects of neurogenesis, thus ensuring that neurons of the correct phenotype are generated in proper numbers at the appropriate times and locations. PMID- 11060245 TI - Synaptogenesis in the giant-fibre system of Drosophila: interaction of the giant fibre and its major motorneuronal target. AB - The tergotrochanteral (jump) motorneuron is a major synaptic target of the Giant Fibre in Drosophila. These two neurons are major components of the fly's Giant Fibre escape system. Our previous work has described the development of the Giant Fibre in early metamorphosis and the involvement of the shaking-B locus in the formation of its electrical synapses. In the present study, we have investigated the development of the tergotrochanteral motorneuron and its electrical synapses by transforming Drosophila with a Gal4 fusion construct containing sequences largely upstream of, but including, the shaking-B(lethal) promoter. This construct drives reporter gene expression in the tergotrochanteral motorneuron and some other neurons. Expression of green fluorescent protein in the motorneuron allows visualization of its cell body and its subsequent intracellular staining with Lucifer Yellow. These preparations provide high resolution data on motorneuron morphogenesis during the first half of pupal development. Dye-coupling reveals onset of gap-junction formation between the tergotrochanteral motorneuron and other neurons of the Giant-Fibre System. The medial dendrite of the tergotrochanteral motorneuron becomes dye-coupled to the peripheral synapsing interneurons between 28 and 32 hours after puparium formation. Dye-coupling between tergotrochanteral motorneuron and Giant Fibre is first seen at 42 hours after puparium formation. All dye coupling is abolished in a shaking-B(neural) mutant. To investigate any interactions between the Giant Fibre and the tergotroachanteral motorneuron, we arrested the growth of the motorneuron's medial neurite by targeted expression of a constitutively active form of Dcdc42. This results in the Giant Fibre remaining stranded at the midline, unable to make its characteristic bend. We conclude that Giant Fibre morphogenesis normally relies on fasciculation with its major motorneuronal target. PMID- 11060246 TI - Delta 1-activated notch inhibits muscle differentiation without affecting Myf5 and Pax3 expression in chick limb myogenesis. AB - The myogenic basic helix-loop-helix (bHLH) transcription factors, Myf5, MyoD, myogenin and MRF4, are unique in their ability to direct a program of specific gene transcription leading to skeletal muscle phenotype. The observation that Myf5 and MyoD can force myogenic conversion in non-muscle cells in vitro does not imply that they are equivalent. In this paper, we show that Myf5 transcripts are detected before those of MyoD during chick limb development. The Myf5 expression domain resembles that of Pax3 and is larger than that of MyoD. Moreover, Myf5 and Pax3 expression is correlated with myoblast proliferation, while MyoD is detected in post-mitotic myoblasts. These data indicate that Myf5 and MyoD are involved in different steps during chick limb bud myogenesis, Myf5 acting upstream of MyoD. The progression of myoblasts through the differentiation steps must be carefully controlled to ensure myogenesis at the right place and time during wing development. Because Notch signalling is known to prevent differentiation in different systems and species, we sought to determine whether these molecules regulate the steps occurring during chick limb myogenesis. Notch1 transcripts are associated with immature myoblasts, while cells expressing the ligands Delta1 and Serrate2 are more advanced in myogenesis. Misexpression of Delta1 using a replication-competent retrovirus activates the Notch pathway. After activation of this pathway, myoblasts still express Myf5 and Pax3 but have downregulated MyoD, resulting in inhibition of terminal muscle differentiation. We conclude that activation of Notch signalling during chick limb myogenesis prevents Myf5 expressing myoblasts from progressing to the MyoD-expressing stage. PMID- 11060247 TI - Nanos interacts with cup in the female germline of Drosophila. AB - Nanos (Nos) is a translational regulator that governs abdominal segmentation of the Drosophila embryo in collaboration with Pumilio (Pum). In the embryo, the mode of Nos and Pum action is clear: they form a ternary complex with critical sequences in the 3'UTR of hunchback mRNA to regulate its translation. Nos also regulates germ cell development and survival in the ovary. While this aspect of its biological activity appears to be evolutionarily conserved, the mode of Nos action in this process is not yet well understood. In this report, we show that Nos interacts with Cup, which is required for normal development of the ovarian germline cells. nos and cup also interact genetically--reducing the level of cup activity specifically suppresses the oogenesis defects associated with the nos(RC) allele. This allele encodes a very low level of mRNA and protein that, evidently, is just below the threshold for normal ovarian Nos function. Taken together, these findings are consistent with the idea that Nos and Cup interact to promote normal development of the ovarian germline. They further suggest that Nos and Pum are likely to collaborate during oogenesis, as they do during embryogenesis. PMID- 11060248 TI - [Inflammation, infection and coronary artery disease: myths and realities. Special XXXV Conference of the National Congress of the Spanish Society of Cardiology]. AB - In the past decades it has become apparent that inflammation plays a role in atherogenesis and rapid coronary artery disease progression. Active, or vulnerable, atheromatous plaques are responsible for acute coronary events and contain high concentrations of inflammatory cells as well as molecules involved in the inflammatory process, such as cytokines, adhesion molecules and growth factors. From a clinical perspective, early detection of these plaques may prevent the occurrence of serious coronary events. Unfortunately, current diagnostic techniques -i.e. angiography- do not allow the characterization of events taking place in the arterial wall. Therefore, these diagnostic tools cannot identify vulnerable plaques. Recent studies have suggested that markers of systemic inflammation may help in the detection of high risk patients. Although the role of inflammation in the pathogenesis of atherosclerosis is established, it is not known what triggers inflammation in this context. Infectious agents such as viruses and Gram negative bacteria -i.e. Chlamydia pneumoniae- have been postulated to play a role. Several mechanisms, involving inflammation and immunological processes, have been suggested to explain how chronic infections may cause atherosclerosis. Small pilot studies have also been carried out which suggest a causal role of infection in coronary artery disease. These results, however, await confirmation by other large, currently ongoing, studies. The infectious hypothesis of atherosclerosis is still a matter of debate; however, this theory has contributed to the rapid advance of our knowledge regarding the pathogenesis of coronary artery disease in the past few years. Moreover, the notion that coronary artery disease can be considered to be an inflammatory condition in its own right has opened new and challenging avenues for research. PMID- 11060249 TI - [Where is atrial fibrillation research going to?]. PMID- 11060250 TI - [Intraoperative transesophageal echocardiography. What for? How? For whom?]. PMID- 11060251 TI - [Influence of pre-infarction angina on mid-term mortality after acute myocardial infarction]. AB - INTRODUCTION AND OBJECTIVES: Pre-infarction angina may reduce the extent of myocardial cell necrosis and improves the prognosis after myocardial infarction. The aim of this study was to analyze the total mortality six-month after acute myocardial infarction according to the presence or absence of pre-infarction angina. METHODS: One hundred seventy-five consecutive patients with acute myocardial infarction were prospectively included, 72 (41.4%) with pre-infarction angina. They were followed for 6 months. There were 16 deaths (15.5%) in the group of patients without pre-infarction angina and 7 (9.7%) in the group with pre-infarction angina (log-rank = 1.03; p = 0.311). The hazard-risk function curves showed a higher risk of death during the entire follow-up in the group without pre-infarction angina. In the multivariate logistic regression model, the presence of pre-infarction angina does not significantly reduce the risk of death (OR = 0.43; CI 95% = 0.09-2. 22; p = 0.303). We detected a significant interaction between treatment with sulfonylureas before the infarction and the presence of pre-infarction angina (p = 0.017). CONCLUSIONS: In this study no significant differences were observed in total mortality six months after acute myocardial infarction according to the presence of pre-infarction angina. However, the risk of death seemed to be higher in the group of patients without pre-infarction angina during the entire follow-up. A significant interaction was found between the treatment with sulfonylurea drugs before infarction and the presence of pre-infarction angina. PMID- 11060252 TI - [Effect of the use of echoenhancers on interobserver variability in dobutamine stress echocardiography]. AB - INTRODUCTION: Dobutamine stress echocardiography is an accurate technique for the noninvasive diagnosis of coronary artery disease. However, interobserver variability is an important limitation of stress echocardiography. Image quality and echocardiographer experience have been described to influence interobserver agreement. AIM: The aim of this study was to determine whether use of contrast agents during dobutamine stress echocardiography improves the agreement between an experienced and a unexperienced observer, and if learning period would be influenced by the use of contrast. METHODS: Two blind observers interpreted all the studies: one experienced echocardiographer (A) and one unexperienced observer (B) in this technique. The contrast agent Levovist/Levograf 2.5 g was administered by two bolus (at rest and at peak stress). In all cases, second harmonic imaging and stress digitalisation packs were used. The kappa test was used to determine interobserver agreement. RESULTS: Fifty-two unselected consecutive studies in 51 patients were analyzed. Twenty-two studies were performed with contrast. The agreement between the experienced and the unexperienced observer was Kappa 0.58 and 0.52, with and without the use of contrast, with no statistically significant difference being archived. CONCLUSIONS: The routine use of contrast provides better although not significant, interobserver agreement. However, this improvement is not sufficient to substitute specific training. PMID- 11060253 TI - [Anatomopathological and biochemical correlation of lesions produced by standard and irrigated 4 mm radiofrequency catheters]. AB - INTRODUCTION AND OBJECTIVES: The influence on the size of radiofrequency lesions by cooling of the tip of the electrode remains unclear. Moreover, the possible effects of two different cooling systems, closed and open, have not been well differentiated. We designed this study to compare both systems of irrigated-tip catheters and the lesions produced with standard 4 mm catheters and also to evaluate the pathological and biochemical marker release correlation (cardiac troponin I) in an experimental model. METHODS: The study was performed in 20 pigs. Applying between 1-8 radiofrequency pulses, at a power of 15, 25 or 50 watts, for 15-60 seconds to each animal. After 7 days, the pigs were sacrificed for anatomopathological study. RESULTS: A total of 54 lesions were produced, 25 with standard catheters and 29 with irrigated catheters. The mean volume of the lesions produced with standard catheters was 146 +/- 110 microl and with irrigated-tip catheters 856 +/- 864 microl (p < 0.001). Peak values of cardiac troponin I were also higher for irrigated catheters (18 +/- 15 ng/ml) than for standard (6.5 +/- 3 ng/ml). The correlation between the size of the lesion and the levels of cardiac troponin I were 0.86 and 0.79 with the standard and irrigated-tip catheters, respectively. The incidence of cratering was higher with standard catheters (60%) than with irrigated (27%). CONCLUSIONS: The lesions produced with an irrigated catheter are greater than those observed with standard catheters. The mean peak value of postablation cardiac troponin demonstrate a good correlation with the real size of the necrosis. PMID- 11060254 TI - [Epicardial mapping of reentrant activation during ventricular fibrillation. An experimental study]. AB - INTRODUCTION AND OBJECTIVES: High-resolution epicardial mapping was used in an experimental model to analyze reentrant activation during ventricular fibrillation. METHODS: In 30 isolated Langendorff-perfused rabbit hearts, recordings were made of ventricular fibrillation activity using an epicardial multiple electrode. In the activation maps with reentrant activation patterns, determinations were made of the number of consecutive rotations, the maximum length of the central core, the area encompassed by the core and two electrodes surrounding it, and the cycle defined by reentrant activation. RESULTS: Most of the activation maps analyzed showed complex patterns with two or more wave fronts that either collided or remained separated by functional block lines (514 maps, 86%). In 112 maps (19%) activation patterns compatible with epicardial breakthrough of the depolarization process were observed. Reentrant activity was recorded in 42 maps (7%) - the maximum number of consecutive rotations being 3 (mean = 1.3 +/- 0.5). The maximum length of the central core ranged from 3 to 7 mm (mean = 5 +/- 1 mm), while the area encompassed by the central core plus two electrodes surrounding it ranged from 35 to 55 mm2 (mean = 45 +/- 6 mm2). The reentrant cycle length (mean = 47 +/- 8 ms) showed a linear relation to the maximum length of the central core reentry (cycle = 4.52 x length + 24.6; r = 0.7; p < 0.0001). CONCLUSIONS: a) Epicardial mapping allowed the identification of reentrant activation patterns during ventricular fibrillation in the experimental model used; b) the reentrant activity detected is infrequent and unstable, and c) a linear relation exists between the duration of the cycles defined by reentrant activity and the maximum length of central core reentry. PMID- 11060255 TI - [Pre and perioperative factors determining early in-hospital mortality in patients over 75 years of age undergoing cardiac surgery]. AB - INTRODUCTION AND OBJECTIVES: To know the in-hospital morbidity and mortality and the related factors in patients over 75 years old undergoing cardiac surgery. PATIENTS AND METHODS: A retrospective analysis was carried out in 252 out of 2043 consecutive patients (129 female, 123 male) over 75 years of age (mean age 77.8 years; range 75 to 89 years) undergoing open heart surgery from january 1, 1994 to november 30, 1997. Isolated aortic valve replacement was performed in 128 patients, 78 underwent isolated coronary artery bypass grafting and 46 combined surgery. Preoperative determinants of morbidity and mortality were analyzed. RESULTS: The overall hospital mortality was 15.1%, 13.2% in the aortic group, 12.8% in the coronary group and 23.9% in the combined surgery group. The overall morbidity rate was 38.6% and 25.8%, 34.2% in the aortic and coronary groups, respectively. Preoperative risk factors were prior surgery (p < 0.0004) and emergency operation (p < 0.04). In aortic valve replacement, NYHA class IV (p < 0.05), prior operation (p < 0. 01) and emergency surgery (p < 0.01) were determinant. Perioperative factors of early mortality were: prolonged cross clamping > 60 min (p < 0.02), cardiopulmonary bypass time > 90 min (p < 0.002), need for inotropic drugs (p < 0.005) and postoperative complications (p < 0.00001). Mean postoperative length of hospital stay was 12.8 +/- 8. 5 days. CONCLUSIONS: Despite the greater rate of early morbimortality in patients over 75 years of age, cardiac surgery may be performed avoiding emergency surgery, functional grade IV and prolonged length of surgery. PMID- 11060256 TI - [Intraoperative transesophageal echocardiography: guidelines for anesthesiologist training. Joint Task Force of the Echocardiography Section and other Imaning Techniques of the Spanish Society of Cardiology and of the Cardiothoracic Surgery Section of the Spanish Society of Anesthesiology, Recovery, and Pain Therapy]. PMID- 11060257 TI - [Practice guidelines of the Spanish Society of Cardiology on endocarditis]. AB - Infectious endocarditis is a disease which mainly involves the cardiac valves. It has a bad prognosis and is caused by a great variety of microorganisms. Prophylaxis is important although the effectiveness and the best way to achieve it remain unclear. Recommendations are herein presented. The diagnosis is based on clinical, bacteriological, and echocardiographic findings mainly based on Duke's criteria. Transthoracic and transesophageal echography are not only of diagnostic value but are also a tool to determine the therapy to follow. Antibiotic therapy should be selected according to the organisms isolated and their in vitro susceptibility. Guidelines for empirical antibiotic therapy in cases of negative cultures are also included. Lastly, indications and time for surgery are discussed. PMID- 11060258 TI - [Anomalous origin of the right coronary artery]. PMID- 11060259 TI - [Single coronary artery and right coronary ostium agenesis]. PMID- 11060260 TI - [Cardioinhibitory vagal response not related to pain, and induced by radiofrequency application during ablation of right posteroseptal accessory pathway]. AB - Inappropriate sinus tachycardia is the most common arrhythmia induced by radiofrequency energy delivery in the posteroseptal area. It has been suggested that this could be secondary to parasymphathetic nerve injury. We report a patient with extreme sinus bradycardia and PR interval prolongation induced by radiofrequency energy delivered in the coronary sinus ostium area, but not related to any other stimulus. The most probable mechanism of the disorder was transient stimulation of the vagal afferent nerve fibers located in this anatomical area. PMID- 11060261 TI - [Mycotic aneurysm caused by Aspergillus of the aortic suture line after heart transplantation]. AB - The first case of mycotic aneurysm of aorta by Aspergillus in a patient with heart transplantation is described, in which the infection was produced by direct surgical contamination of the aortic suture. The period of latency was of eight months. The unusualness of the case and its diagnostic difficulties, are is commented. PMID- 11060262 TI - [The bidirectional cavopulmonary (Glenn) shunt without cardiopulmonary bypass: a safe and advisable technique]. AB - The bidirectional cavopulmonary (Glenn) shunt is almost a routine first step procedure for total cavopulmonary connection in children with single-ventricle cardiac anomalies. It is usually performed with cardiopulmonary bypass, of which adverse effects can be especially deleterious in these cardiac conditions. To avoid these adverse effects, we performed the cavopulmonary shunt in 5 children through sternotomy without cardiopulmonary bypass. There was no mortality nor morbidity. We think that this technique is safe, reproducible, and even advisable in children with single-ventricle anomalies. PMID- 11060263 TI - [Oblique orientation of the accessory pathway demonstrated by radiofrequency application]. AB - Activation mapping of atrial and ventricular insertion has suggested an oblique orientation of some accessory pathways. However, this aspect has not been demonstrated by radiofrequency application. This report presents two patients with Wolff-Parkinson-White syndrome and an accessory pathway with bidirectional conduction and oblique orientation. The accessory pathway oblique orientation was demonstrated by transient and permanent conduction abolition following radiofrequency application in two separate ventricular and atrial sites. These findings may explain the failure to ablate accessory pathway by radiofrequency application in the ventricular side of the mitral annulus guided by retrograde atrial activation occasionally observed in patients with a concealed accessory pathway. PMID- 11060264 TI - [Myocardial scintigraphy and patency of coronary bypass graft with mammary artery]. PMID- 11060266 TI - [ [In Process Citation] PMID- 11060265 TI - [Exercise treadmill test in patients with unstable angina]. PMID- 11060267 TI - [ [In Process Citation] PMID- 11060268 TI - [The National Committee of our specialty]. PMID- 11060269 TI - [Minimally invasive parathyroid surgery with 99mTc-sestamibi scintigraphy and probe-radioguided surgery: preliminary results]. AB - OBJECTIVE: The most common pathological finding in patients with sporadic primary hyperparathyroidism is a single adenoma. The success of parathyroid surgery is determined by the identification and removal of the responsible adenoma. The aim of our study was to minimize interventional techniques, hospital stay and overall costs of patients with parathyroid adenoma who do not require a complete neck examination. METHODS: Seven patients with biochemically confirmed hyperparathyroidism were included in our study. All the patients underwent 99mTc Tetrofosmin and 99mTc-Pertecnetate neck scans in the days prior to the surgery. On the day of the operation, we administrated a dose of 740-925 MBq 99mTc sestamibi. The scintigraphy study and radioguided surgery examination were performed with a 10 mm hand-held gamma probe at 2-3 hours of the administration. The counts were measured in the four neck quadrants. RESULTS: We identified seven adenomas and six were removed with a 2 cm incision. The removal of the adenoma resulted in a decline in radioactivity in that quadrant, these being comparable in all the neck quadrants. Ex vivo counts identified parathyroid tissue from fat and lymph node. The histopathologic exam and a decrease of the intact PTHi levels at 30 and 60 minutes after removal of the adenoma confirmed the correct surgery. CONCLUSIONS: Radioguided parathyroid surgery allows for a minimally invasive unilateral neck examination with a small scar and better cosmetic results, reduces operative and anesthetic times, requires minimal times spent in the hospital and achieves a reduction in overall cost. PMID- 11060270 TI - [Bone scintigraphy findings in patients with recently diagnosed adenocarcinoma of the prostate: relationship with prostate specific antigen levels]. AB - BACKGROUND AND OBJECTIVES: Although it has been suggested that bone scintigraphy (BS) may not be necessary in patients with prostate adenocarcinoma (PA) and normal prostate specific antigen (PSA) plasma levels, controversy still remains. The objective of the study was to evaluate the existing relationship between PSA plasma levels and BS findings in patients with a recently diagnosed PA in order to assess whether BS may be omitted on the basis of the PSA levels in these patients. METHODS: The 475 patients (70+/-7 years old) consecutively diagnosed of PA between 1994 and 1998 in our institution made up the study population. PSA plasmatic levels were determined and BS was performed (body planar study after 99mTc-methyl diphosphonate 900 MBq administration) in all the patients. RESULTS: In 362 patients (76.2%), BS was negative, in 108 (22.7%) positive and undetermined in the remaining 5 patients (1.1%). The mean PSA level in the whole study population was 74+/-267 ng/ml (range 0.4-4.200) and was higher in patients with positive GO (218+/-512 vs 31+/-89, p<0,0001). As PSA increased, the rate of patients with positive BS was significantly higher, this being 0%, 16.4%, 9.7%, 14.0%, 31.0% and 48.3% in patients with PSA 4, 4.1-10; 10.1-20; 20.1-30; 30.1-40, and >40 ng/ml, respectively (p<0.0001). In patients with positive BS, the PSA levels were 20 and 10 ng/ml in 30.6% and 18.6% of the patients, respectively. CONCLUSION: According to our experience, there is a significant association between PSA plasma levels and the BS results in patients with recently diagnosed PA. However, a significant proportion of patients with bone metastasis have normal PSA levels, and therefore BS should be performed in all patients with recently diagnosed PA regardless of the PSA levels. PMID- 11060271 TI - [Scintigraphy with 99mTc-MIBI in the diagnosis of axillary lymph node invasion of breast cancer]. AB - The presence of affected locoregional lymph nodes should be considered as one of the most important prognostic factors of breast cancer. At present, the clinician is conditioned by an absolute lack of an efficient methodology to evaluate the possible invasion of the axillary lymph nodes, which if negative, would make it possible to avoid surgical excision. In this study, we will evaluate the use of the 99mTc-MIBI scintigraphy in the pre-surgical diagnosis of axillary lymph node invasion and will analyze the relationship between the 99mTc-MIBI uptake and the number of lymph nodes affected. MATERIAL AND METHODS: 84 patients diagnosed of breast cancer were analyzed in this study. All of them underwent a 99mTc-MIBI scintigraphy, and the tumor/background ratio was determined semiquantitively for each image. The axillary lymph node invasion was determined following surgery. RESULTS: The sensitivity of the breast scintigraphy with 99mTc-MIBI for detection of lymph node invasion is 36% and the specificity is 100%. The positive predictive value is 100% and the negative one 48%. In the current study, we failed to detect correlation between the intensity of 99mTc-MIBI uptake in the primary tumor and the number of affected axillary lymph nodes. CONCLUSION: 99mTc MIBI breast scintigraphy can provide complementary information for the presurgical diagnosis of breast cancer axillary lymph node invasion. 99mTc-MIBI breast scintigraphy shows high specificity and a high predictive value. PMID- 11060272 TI - The effect of aminophylline administration on 99mTc-MIBI lung and liver uptake in patients with or without myocardial ischemia. AB - This work aims to analyze the influence of aminophylline in the pulmonary and hepatic uptake of 99mTc-methoxyisobutil isonitrile (99mTc-MIBI). 72 patients were studied and a myocardial perfusion (MPS) single photon emission computed tomography (SPECT) with 99mTc-MIBI was carried out after the administration of dipyridamole. According to the MPS, the patients were classified into 2 groups: Group A: 45 patients without myocardial ischemia and Group B: 27 with ischemia. Each group was divided into 2 subgroups according to whether they had (I) or had not (II) received intravenous aminophylline. The dipyridamole was administered for 4 minutes at a dose of 0.56 mg/kg. If the patients presented any complication, intravenous aminophylline was administered. At 30 minutes p.i., planar images were obtained during a scintigraphy in the interior projection after the injection of 99mTc-MIBI. The regions of interest in the heart, hepatic cupula, and most active area of the left lung were outlines and the activity rates were calculated: lung/heart (LHR) and liver/heart (LivHR). No statistically significant differences were observed in the uptake of 99mTc-MIBI between subgroups I and II. However, the LHR rates in both subgroups were significantly lower in the patients with normal myocardial perfusion than in the patients with ischemia: LHR group A1 vs B1: 0. 32 +/- 0.08 vs 0.36 +/- 0.06, p = 0.03; group AII vs BII 0.31 +/- 0. 07 vs 0.35 +/- 0.07, p = 0.01 respectively. In conclusion, the administration of aminophylline, after the infusion of dipyridamole for MPS, does not modify the pulmonary or hepatic uptake of 99mTc-MIBI. PMID- 11060273 TI - [Clinico-biological impact of pS2 positivity in estrogen receptor negative infiltrating ductal carcinomas of the breast]. AB - The pS2 protein is regulated by estrogens, but it can also be expressed in hormone independent breast carcincomas. We have carried out the present study in order to analyze the clinical-biological impact of pS2 positivity (>2 ng/mg prt.) in negative estrogen receptors (< 10 fmol/mg prt.) infiltrating ductal breast carcinomas (IDC). MATERIAL AND METHODS: 97 negative ER-IDC have been included in our study. We established the doses of the cytosol levels of pS2, progesterone receptors (PR), cathepsin D, tissue -type plasminogen activator (t-PA) and hyaluronic acid (HA), as well as the levels of HA, epidermal growth factor receptor (EGFR), CD44v5 and CD44v6 in cell surface membranes. We also considered the menopausal status, histological grade, ploidy, cellular synthesis phase, tumor size, axillary lymph node involvement and the existence of distant metastasis. The same results were obtained when the progesterone receptor status was also considered. RESULTS: ER-/pS2+ IDC presented higher (p <0,05) PR, t-PA and HA cytosol level, as well as lower EGFR concentrations, S-phase > 7% and S phase >14% and lower N+>10 percentages and aneuploidy. They were also more frequently CD44v6+. The same results were observed when the positivity of the progesterone receptors was considered. CONCLUSIONS: The above results lead us to consider that the positivity for pS2 in ER- IDC is associated with hormone dependent parameters, good differentiation and lower cellular proliferation, which can explain a better clinical outcome. PMID- 11060274 TI - [Alteration in the biodistribution of 99mTc-albumin microspheres after its administration through a canalized umbilical vein]. AB - We present the case of a newborn female patient with a suspected right hypoplastic lung who was referred to our Service to perform a perfusion lung scintigraphy. The tracer (99mTc-albumin microspheres) was injected with a central catheter through the umbilical vein. The lung scan showed abnormal extrapulmonary activity that was attributed to an erroneous canalization (catheter in Arantius duct). A new study, with injection through the peripheral vein, showed the lung perfusion, with a global decrease of activity in the right lung. This case demonstrates a cause of abnormal extrapulmonary perfusion tracer uptake, verifying that using the correct access route is mandatory. PMID- 11060275 TI - State of the art of therapy in nuclear medicine. PMID- 11060276 TI - [Hepatosplenic scintigraphy in a case of multiple abdominal splenosis]. PMID- 11060277 TI - [Scintigraphy with 99mTc-HMPAO-marked leukocytes in toxic megacolon]. PMID- 11060278 TI - [Informed consent in radioisotopic treatment of metastatic bone pain. Sociedad Espanola de Medicina Nuclear]. PMID- 11060281 TI - Rational design of a potent anticoagulant thrombin. AB - Thrombin acts as a procoagulant when it cleaves fibrinogen and promotes the formation of a fibrin clot and functions as an anticoagulant when it activates protein C with the assistance of the cofactor thrombomodulin. The dual function of thrombin in the blood poses the challenge to turn the enzyme into a potent anticoagulant by selectively abrogating fibrinogen cleavage. Using functional and structural data, we have rationally designed a thrombin mutant, W215A/E217A, that cleaves fibrinogen with a value of k(cat)/K(m) about 20,000-fold slower than wild type but activates protein C in the presence of thrombomodulin with a specificity comparable with wild-type. This mutant demonstrates for the first time that the relative specificity of thrombin toward fibrinogen and protein C can be completely reversed. PMID- 11060282 TI - Molecular cloning of a novel scavenger receptor for oxidized low density lipoprotein, SR-PSOX, on macrophages. AB - Receptor-mediated endocytosis of oxidized low density lipoprotein (OxLDL) by macrophages has been implicated in foam cell transformation in the process of atherogenesis. Although several scavenger receptor molecules, including class A scavenger receptors and CD36, have been identified as OxLDL receptors on macrophages, additional molecules on macrophages may also be involved in the recognition of OxLDL. From a cDNA library of phorbol 12-myristate 13-acetate stimulated THP-1 cells, we isolated a cDNA encoding a novel protein designated SR PSOX (scavenger receptor that binds phosphatidylserine and oxidized lipoprotein), which acts as a receptor for OxLDL. SR-PSOX was a type I membrane protein consisting of 254 amino acids, expression of which was shown on human and murine macrophages with a molecular mass of 30 kDa. SR-PSOX could specifically bind with high affinity, internalize, and degrade OxLDL. The recognition of OxLDL was blocked by polyinosinic acid and dextran sulfate but not by acetylated low density lipoprotein. Taken together, SR-PSOX is a novel class of molecule belonging to the scavenger receptor family, which may play important roles in pathophysiology including atherogenesis. PMID- 11060283 TI - Heterogeneity within animal thioredoxin reductases. Evidence for alternative first exon splicing. AB - Animal thioredoxin reductases (TRs) are selenocysteine-containing flavoenzymes that utilize NADPH for reduction of thioredoxins and other protein and nonprotein substrates. Three types of mammalian TRs are known, with TR1 being a cytosolic enzyme, and TR3, a mitochondrial enzyme. Previously characterized TR1 and TR3 occurred as homodimers of 55-57-kDa subunits. We report here that TR1 isolated from mouse liver, mouse liver tumor, and a human T-cell line exhibited extensive heterogeneity as detected by electrophoretic, immunoblot, and mass spectrometry analyses. In particular, a 67-kDa band of TR1 was detected. Furthermore, a novel form of mouse TR1 cDNA encoding a 67-kDa selenoprotein subunit with an additional N-terminal sequence was identified. Subsequent homology analyses revealed three distinct isoforms of mouse and rat TR1 mRNA. These forms differed in 5' sequences that resulted from the alternative use of the first three exons but had common downstream sequences. Similarly, expression of multiple mRNA forms was observed for human TR3 and Drosophila TR. In these genes, alternative first exon splicing resulted in the formation of predicted mitochondrial and cytosolic proteins. In addition, a human TR3 gene overlapped with the gene for catechol-O methyltransferase (COMT) on a complementary DNA strand, such that mitochondrial TR3 and membrane-bound COMT mRNAs had common first exon sequences; however, transcription start sites for predicted cytosolic TR3 and soluble COMT forms were separated by approximately 30 kilobases. Thus, this study demonstrates a remarkable heterogeneity within TRs, which, at least in part, results from evolutionary conserved genetic mechanisms employing alternative first exon splicing. Multiple transcription start sites within TR genes may be relevant to complex regulation of expression and/or organelle- and cell type-specific location of animal thioredoxin reductases. PMID- 11060284 TI - tRNA recognition of tRNA-guanine transglycosylase from a hyperthermophilic archaeon, Pyrococcus horikoshii. AB - In the biosynthesis of archaeosine, archaeal tRNA-guanine transglycosylase (TGT) catalyzes the replacement of guanine at position 15 in the D loop of most tRNAs by a free precursor base. We examined the tRNA recognition of TGT from a hyperthermophilic archaeon, Pyrococcus horikoshii. Mutational studies using variant tRNA(Val) transcripts revealed that both guanine and its location (position 15) were strictly recognized by TGT without any other sequence-specific requirements. It appeared that neither the global L-shaped structure of a tRNA nor the local conformation of the D loop contributed to recognition by TGT. A minihelix composed of the acceptor stem and D arm of tRNA(Val), designed as a potential minimal substrate, failed to serve as a substrate for TGT. Only a minihelix with mismatched nucleotides at the junction between the two domains served as a good substrate, suggesting that mismatched nucleotides in the helix provide the specific information that allows TGT to recognize the guanine in the D loop. Our findings indicate that the tRNA recognition requirements of P. horikoshii TGT are sufficiently limited and specific to allow the enzyme to recognize efficiently any tRNA species whose structure is not fully stabilized in an extremely high temperature environment. PMID- 11060285 TI - Solution structure and dynamics of myeloid progenitor inhibitory factor-1 (MPIF 1), a novel monomeric CC chemokine. AB - MPIF-1, a CC chemokine, is a specific inhibitor of myeloid progenitor cells and is the most potent activator of monocytes. The solution structure of myeloid progenitor inhibitor factor-1 (MPIF-1) has been determined by NMR spectroscopy. The structure reveals that MPIF-1 is a monomer with a well defined core except for termini residues and adopts the chemokine fold of three beta-strands and an overlying alpha-helix. In addition to the four cysteines that characterize most chemokines, MPIF-1 has two additional cysteines that form a disulfide bond. The backbone dynamics indicate that the disulfide bonds and the adjacent residues that include the functionally important N-terminal and N-terminal loop residues show significant dynamics. MPIF-1 is a highly basic protein (pI >9), and the structure reveals distinct positively charged pockets that could be correlated to proteoglycan binding. MPIF-1 is processed from a longer proprotein at the N terminus and the latter is also functional though with reduced potency, and both proteins exist as monomers under a variety of solution conditions. MPIF-1 is therefore unique because longer proproteins of all other chemokines oligomerize in solution. The MPIF-1 structure should serve as a template for future functional studies that could lead to therapeutics for preventing chemotherapy associated myelotoxicity. PMID- 11060286 TI - Induction of rapid histone degradation by the cytotoxic T lymphocyte protease Granzyme A. AB - The cytotoxic T lymphocyte protease granzyme A induces caspase-independent cell death in which DNA single-strand nicking is observed instead of oligonucleosomal fragmentation. Granzyme A is a specific tryptase that concentrates in the nucleus of targeted cells and synergistically enhances DNA fragmentation induced by the caspase activator granzyme B. Here we show that granzyme A treatment of isolated nuclei enhances DNA accessibility to exogenous endonucleases. In vitro and after cell loading with perforin, GrnA completely degrades histone H1 and cleaves core histones into approximately 16-kDa fragments. Histone digestion provides a mechanism for unfolding compacted chromatin and facilitating endogenous DNase access to DNA during T cell and natural killer cell granule-mediated apoptosis. PMID- 11060287 TI - Unique protein determinants of the subtype-selective ligand responses of the estrogen receptors (ERalpha and ERbeta ) at AP-1 sites. AB - The two subtypes of human estrogen receptor, alpha (hERalpha) and beta (hERbeta), regulate transcription at an AP-1 response element differently in response to estradiol and the anti-estrogens tamoxifen and raloxifene. To better understand the protein determinants of these differences, chimeric and deletional mutants of the N-terminal domain and the F region of ERalpha and ERbeta were made and tested in transient transfection assays at the classical estrogen response element (ERE) site as well as at an AP-1 site. Although the same regions on each receptor subtype appeared to be primarily responsible for estradiol activation at an ERE and in HeLa cells, major differences between ERalpha and ERbeta mutants were seen in the estrogen and anti-estrogen responses at an AP-1 site. This differential ligand response maps to the N-terminal domain and the F region. These results suggest that different estrogenic and anti-estrogenic ligands use different mechanisms of activation and inhibition at the AP-1 site. In contrast to previous studies, this work also shows that many of subtype-specific responses are not transferred to the other subtype by swapping the N-terminal domain of the receptor. This implies that there are other unique surfaces presented by each subtype outside of the N-terminal domain, and these surfaces can play a role in subtype-selective signaling. Together, these data suggest a complex interface between ligand, response element, and receptor that underlies ligand activation in estrogen signaling pathways. PMID- 11060288 TI - Olfactory receptor trafficking involves conserved regulatory steps. AB - Olfactory receptors are difficult to functionally express in heterologous cells. They are typically retained in the endoplasmic reticulum of cells commonly used for functional expression studies and are only released to the plasma membrane in mature cells of the olfactory receptor neuron lineage. A recently developed olfactory cell line, odora, traffics olfactory receptors to the plasma membrane when differentiated. We found that undifferentiated odora cells do not traffic olfactory receptors to their surface, even though they release the receptors to the Golgi apparatus and endosomes. This behavior differs from other cell lines tested thus far. Differentiated odora cells also properly traffic vomeronasal receptors of the VN1 type, which lack sequence similarity to olfactory receptors. ODR-4, a protein that is necessary for plasma membrane trafficking of a chemosensory receptor in nematodes, facilitates trafficking of rat olfactory receptor U131 in odora and Chinese hamster ovary cells. Olfactory receptor trafficking from the endoplasmic reticulum to the plasma membrane involves at least two steps whose regulation depends on the maturation state of cells in the olfactory receptor neuron lineage. These results also indicate that some components of the regulatory mechanism are conserved. PMID- 11060289 TI - Rho GTPases as modulators of the estrogen receptor transcriptional response. AB - The estrogen receptor alpha (ER) is a ligand-dependent transcription factor that plays a critical role in the development and progression of breast cancer, in part, by regulating target genes involved in cellular proliferation. To identify novel components that affect the ER transcriptional response, we performed a genetic screen in yeast and identified RDI1, a Rho guanine nucleotide dissociation inhibitor (Rho GDI), as a positive regulator of ER transactivation. Overexpression of the human homologue of RDI1, Rho GDIalpha, increases ERalpha, ERbeta, androgen receptor, and glucocorticoid receptor transcriptional activation in mammalian cells but not activation by the unrelated transcription factors serum response factor and Sp1. In contrast, expression of constitutively active forms of RhoA, Rac1, and Cdc42 decrease ER transcriptional activity, suggesting that Rho GDI increases ER transactivation by antagonizing Rho function. Inhibition of RhoA by expression of either the Clostridium botulinum C3 transferase or a dominant negative RhoA resulted in enhanced ER transcriptional activation, thus phenocopying the effect of Rho GDI expression on ER transactivation. Together, these findings establish the Rho GTPases as important modulators of ER transcriptional activation. Since Rho GTPases regulate actin polymerization, our findings suggest a link between the major regulators of cellular architecture and steroid receptor transcriptional response. PMID- 11060290 TI - The role of G protein activation in the toxicity of amyloidogenic Abeta-(1-40), Abeta-(25-35), and bovine calcitonin. AB - More than 16 different proteins have been identified as amyloid in clinical diseases; among these, beta-amyloid (Abeta) of Alzheimer's disease is the best characterized. In the present study, we performed experiments with Abeta and calcitonin, another amyloid-forming peptide, to examine the role of G protein activation in amyloid toxicity. We demonstrated that the peptides, when prepared under conditions that promoted beta-sheet and amyloid fibril (or protofibril) formation, increased high affinity GTPase activity, but the nonamyloidogenic peptides had no discernible effects on GTP hydrolysis. These increases in GTPase activity were correlated to toxicity. In addition, G protein inhibitors significantly reduced the toxic effects of the amyloidogenic Abeta and calcitonin peptides. Our results further indicated that the amyloidogenic peptides significantly increased GTPase activity of purified Galpha(o) and Galpha(i) subunits and that the effect was not receptor-mediated. Collectively, these results imply that the amyloidogenic structure, regardless of the actual peptide or protein sequence, may be sufficient to cause toxicity and that toxicity is mediated, at least partially, through G protein activation. Our abilities to manipulate G protein activity may lead to novel treatments for Alzheimer's disease and the other amyloidoses. PMID- 11060291 TI - Characterization of the two eIF4A-binding sites on human eIF4G-1. AB - Eukaryotic translation initiation factor 4G-1 (eIF4G) plays a critical role in the recruitment of mRNA to the 43 S preinitiation complex. eIF4G has two binding sites for the RNA helicase eIF4A, one in the central domain and one in the COOH terminal domain. Recombinant eIF4G fragments that contained each of these sites separately bound eIF4A with a 1:1 stoichiometry, but fragments containing both sites bound eIF4A with a 1:2 stoichiometry. eIF3 did not interfere with eIF4A binding to the central site. Interestingly, at the same concentration of free eIF4A, more eIF4A was bound to an eIF4G fragment containing both eIF4A sites than the sum of binding to fragments containing the single sites, indicating cooperative binding. Binding of eIF4A to an immobilized fragment of eIF4G containing the COOH-terminal site was competed by a soluble eIF4G fragment containing the central site, indicating that a single eIF4A molecule cannot bind simultaneously to both sites. The association rate constant, dissociation rate constant, and dissociation equilibrium constant for each site were determined by surface plasmon resonance and found to be, respectively, 1.2 x 10(5) m(-1) s(-1), 2.1 x 10(-3) s(-1), and 17 nm for the central site and 5.1 x 10(3) m(-1) s(-1), 1.7 x 10(-3) s(-1), and 330 nm for the COOH-terminal site. PMID- 11060292 TI - Crystal structure of fibroblast growth factor 9 reveals regions implicated in dimerization and autoinhibition. AB - Fibroblast growth factors (FGFs) constitute a large family of heparin-binding growth factors with diverse biological activities. FGF9 was originally described as glia-activating factor and is expressed in the nervous system as a potent mitogen for glia cells. Unlike most FGFs, FGF9 forms dimers in solution with a K(d) of 680 nm. To elucidate the molecular mechanism of FGF9 dimerization, the crystal structure of FGF9 was determined at 2.2 A resolution. FGF9 adopts a beta trefoil fold similar to other FGFs. However, unlike other FGFs, the N- and C terminal regions outside the beta-trefoil core in FGF9 are ordered and involved in the formation of a 2-fold crystallographic dimer. A significant surface area (>2000 A(2)) is buried in the dimer interface that occludes a major receptor binding site of FGF9. Thus, we propose an autoinhibitory mechanism for FGF9 that is dependent on sequences outside of the beta-trefoil core. Moreover, a model is presented providing a molecular basis for the preferential affinity of FGF9 toward FGFR3. PMID- 11060293 TI - The reactive oxygen species--and Michael acceptor-inducible human aldo-keto reductase AKR1C1 reduces the alpha,beta-unsaturated aldehyde 4-hydroxy-2-nonenal to 1,4-dihydroxy-2-nonene. AB - The human aldo-keto reductase AKR1C1 (20alpha(3alpha)-hydroxysteroid dehydrogenase) is induced by electrophilic Michael acceptors and reactive oxygen species (ROS) via a presumptive antioxidant response element (Burczynski, M. E., Lin, H. K., and Penning, T. M. (1999) Cancer Res. 59, 607-614). Physiologically, AKR1C1 regulates progesterone action by converting the hormone into its inactive metabolite 20alpha-hydroxyprogesterone, and toxicologically this enzyme activates polycyclic aromatic hydrocarbon trans-dihydrodiols to redox-cycling o-quinones. However, the significance of its potent induction by Michael acceptors and oxidative stress is unknown. 4-Hydroxy-2-nonenal (HNE) and other alpha,beta unsaturated aldehydes produced during lipid peroxidation were reduced by AKR1C1 with high catalytic efficiency. Kinetic studies revealed that AKR1C1 reduced HNE (K(m) = 34 microm, k(cat) = 8.8 min(-1)) with a k(cat)/K(m) similar to that for 20alpha-hydroxysteroids. Six other homogeneous recombinant AKRs were examined for their ability to reduce HNE. Of these, AKR1C1 possessed one of the highest specific activities and was the only isoform induced by oxidative stress and by agents that deplete glutathione (ethacrynic acid). Several hydroxysteroid dehydrogenases of the AKR1C subfamily catalyzed the reduction of HNE with higher activity than aldehyde reductase (AKR1A1). NMR spectroscopy identified the product of the NADPH-dependent reduction of HNE as 1,4-dihydroxy-2-nonene. The K(m) of recombinant AKR1C1 for nicotinamide cofactors (K(m) NADPH approximately 6 microm, K(m)(app) NADH >6 mm) suggested that it is primed for reductive metabolism of HNE. Isoform-specific reverse transcription-polymerase chain reaction showed that exposure of HepG2 cells to HNE resulted in elevated levels of AKR1C1 mRNA. Thus, HNE induces its own metabolism via AKR1C1, and this enzyme may play a hitherto unrecognized role in a response mounted to counter oxidative stress. AKRs represent alternative GSH-independent/NADPH-dependent routes for the reductive elimination of HNE. Of these, AKR1C1 provides an inducible cytosolic barrier to HNE following ROS exposure. PMID- 11060294 TI - Abnormal contractile function in transgenic mice expressing a familial hypertrophic cardiomyopathy-linked troponin T (I79N) mutation. AB - This study characterizes a transgenic animal model for the troponin T (TnT) mutation (I79N) associated with familial hypertrophic cardiomyopathy. To study the functional consequences of this mutation, we examined a wild type and two I79N-transgenic mouse lines of human cardiac TnT driven by a murine alpha-myosin heavy chain promoter. Extensive characterization of the transgenic I79N lines compared with wild type and/or nontransgenic mice demonstrated: 1) normal survival and no cardiac hypertrophy even with chronic exercise; 2) large increases in Ca(2+) sensitivity of ATPase activity and force in skinned fibers; 3) a substantial increase in the rate of force activation and an increase in the rate of force relaxation; 4) lower maximal force/cross-sectional area and ATPase activity; 5) loss of sensitivity to pH-induced shifts in the Ca(2+) dependence of force; and 6) computer simulations that reproduced experimental observations and suggested that the I79N mutation decreases the apparent off rate of Ca(2+) from troponin C and increases cross-bridge detachment rate g. Simulations for intact living fibers predict a higher basal contractility, a faster rate of force development, slower relaxation, and increased resting tension in transgenic I79N myocardium compared with transgenic wild type. These mechanisms may contribute to mortality in humans, especially in stimulated contractile states. PMID- 11060295 TI - Subcytotoxic H2O2 stress triggers a release of transforming growth factor-beta 1, which induces biomarkers of cellular senescence of human diploid fibroblasts. AB - Stress-induced premature senescence (SIPS) is induced 3 days after exposure of human diploid fibroblasts to subcytotoxic oxidative stress with H(2)O(2), with appearance of several biomarkers of replicative senescence. In this work, we show that transforming growth factor-beta1 (TGF-beta1) regulates the induction of several of these biomarkers in SIPS: cellular morphology, senescence-associated beta-galactosidase activity, increase in the steady-state level of fibronectin, apolipoprotein J, osteonectin, and SM22 mRNA. Indeed, the neutralization of TGF beta1 or its receptor (TGF-beta RII) using specific antibodies decreases sharply the percentage of cells positive for the senescent-associated beta-galactosidase activity and displaying a senescent morphology. In the presence of each of these antibodies, the steady-state level of fibronectin, osteonectin, apolipoprotein J, and SM22 mRNA is no more increased at 72 h after stress. Results obtained on fibroblasts retrovirally transfected with the human papillomavirus E7 cDNA suggest that retinoblastoma protein (Rb) regulates the expression of TGF-beta1 in stressful conditions, leading to SIPS and overexpression of these four genes. PMID- 11060296 TI - Cleavage of the amino terminus of the prion protein by reactive oxygen species. AB - Relatively limited information is available on the processing and function of the normal cellular prion protein, PrP(C). Here it is reported for the first time that PrP(C) undergoes a site-specific cleavage of the octapeptide repeat region of the amino terminus on exposure to reactive oxygen species. This cleavage was both copper- and pH-dependent and was retarded by the presence of other divalent metal ions. The oxidative state of the cell also decreased detection of full length PrP(C) and increased detection of amino-terminally fragmented PrP(C) within cells. Such a post-translational modification has vast implications for PrP(C), in its processing, because such cleavage could alter further proteolysis, and in the formation of the scrapie isoform of the prion protein, PrP(Sc), because abnormal cleavage of PrP(Sc) occurs into the octapeptide repeat region. PMID- 11060297 TI - Polyhydroxybenzoates inhibit ascorbic acid activation of mitochondrial glycerol-3 phosphate dehydrogenase: implications for glucose metabolism and insulin secretion. AB - Glycerol-3-phosphate dehydrogenase from pig brain mitochondria was stimulated 2.2 fold by the addition of 50 microm l-ascorbic acid. Enzyme activity, dependent upon the presence of l-ascorbic acid, was inhibited by lauryl gallate, propyl gallate, protocatechuic acid ethyl ester, and salicylhydroxamic acid. Homogeneous pig brain mitochondrial glycerol-3-phosphate dehydrogenase was activated by either 150 microm L-ascorbic acid (56%) or 300 microm iron (Fe(2+) or Fe(3+) (62%)) and 2.6-fold by the addition of both L-ascorbic acid and iron. The addition of L-ascorbic acid and iron resulted in a significant increase of k(cat) from 21.1 to 64.1 s(-1), without significantly increasing the K(m) of L-glycerol 3-phosphate (10.0-14.5 mm). The activation of pure glycerol-3-phosphate dehydrogenase by either L-ascorbic acid or iron or its combination could be totally inhibited by 200 microm propyl gallate. The metabolism of [5-(3)H]glucose and the glucose-stimulated insulin secretion from rat insulinoma cells, INS-1, were effectively inhibited by 500 microm or 1 mm propyl gallate and to a lesser extent by 5 mm aminooxyacetate, a potent malate-aspartate shuttle inhibitor. The combined data support the conclusion that l-ascorbic acid is a physiological activator of mitochondrial glycerol-3-phosphate dehydrogenase, that the enzyme is potently inhibited by agents that specifically inhibit certain classes of di-iron metalloenzymes, and that the enzyme is chiefly responsible for the proximal signal events in INS-1 cell glucose-stimulated insulin release. PMID- 11060298 TI - Activation of Rac1 and the p38 mitogen-activated protein kinase pathway in response to all-trans-retinoic acid. AB - Several signaling pathways are activated by all-trans-retinoic acid (RA) to mediate induction of differentiation and apoptosis of malignant cells. In the present study we provide evidence that the p38 MAP kinase pathway is activated in a RA-dependent manner in the NB-4, acute pro-myelocytic leukemia, and the MCF-7, breast carcinoma, cell lines. RA treatment of cells induces a time- and dose dependent phosphorylation of p38, and such phosphorylation results in activation of its catalytic domain. p38 activation is not inducible by RA in a variant NB-4 cell line, NB-4.007/6, which is resistant to the effects of RA, suggesting a role for this pathway in the induction of RA responses. Our data also demonstrate that the small G-protein Rac1 is activated by RA and functions as an upstream regulator of p38 activation, whereas the MAPKAPK-2 serine kinase is a downstream effector for the RA-activated p38. To obtain information on the functional role of the Rac1/p38/MAPKAPK-2 pathway in RA signaling, the effects of pharmacological inhibition of p38 on RA-induced gene transcription and cell differentiation were determined. Our results indicate that treatment of cells with the SB203580 inhibitor does not inhibit RA-dependent gene transcription via retinoic acid response elements or induction of Stat1 protein expression. However, treatment with SB203580 or SB202190 strongly enhances RA-dependent induction of cell differentiation and RA-regulated growth inhibitory responses. Altogether, our findings demonstrate that the Rac1/p38 MAP kinase pathway is activated in a RA dependent manner and exhibits negative regulatory effects on the induction of differentiation. PMID- 11060299 TI - Threonine 180 is required for G-protein-coupled receptor kinase 3- and beta arrestin 2-mediated desensitization of the mu-opioid receptor in Xenopus oocytes. AB - To determine the sites in the mu-opioid receptor (MOR) critical for agonist dependent desensitization, we constructed and coexpressed MORs lacking potential phosphorylation sites along with G-protein activated inwardly rectifying potassium channels composed of K(ir)3.1 and K(ir)3.4 subunits in Xenopus oocytes. Activation of MOR by the stable enkephalin analogue, [d Ala(2),MePhe(4),Glyol(5)]enkephalin, led to homologous MOR desensitization in oocytes coexpressing both G-protein-coupled receptor kinase 3 (GRK3) and beta arrestin 2 (arr3). Coexpression with either GRK3 or arr3 individually did not significantly enhance desensitization of responses evoked by wild type MOR activation. Mutation of serine or threonine residues to alanines in the putative third cytoplasmic loop and truncation of the C-terminal tail did not block GRK/arr3-mediated desensitization of MOR. Instead, alanine substitution of a single threonine in the second cytoplasmic loop to produce MOR(T180A) was sufficient to block homologous desensitization. The insensitivity of MOR(T180A) might have resulted either from a block of arrestin activation or arrestin binding to MOR. To distinguish between these alternatives, we expressed a dominant positive arrestin, arr2(R169E), that desensitizes G protein-coupled receptors in an agonist-dependent but phosphorylation-independent manner. arr2(R169E) produced robust desensitization of MOR and MOR(T180A) in the absence of GRK3 coexpression. These results demonstrate that the T180A mutation probably blocks GRK3- and arr3-mediated desensitization of MOR by preventing a critical agonist-dependent receptor phosphorylation and suggest a novel GRK3 site of regulation not yet described for other G-protein-coupled receptors. PMID- 11060300 TI - Phosphatidic acid and diacylglycerol directly activate NADPH oxidase by interacting with enzyme components. AB - The enzyme NADPH oxidase is regulated by phospholipase D in intact neutrophils and is activated by phosphatidic acid (PA) plus diacylglycerol (DG) in cell-free systems. We showed previously that cell-free NADPH oxidase activation by these lipids involves both protein kinase-dependent and -independent pathways. Here we demonstrate that only the protein kinase-independent pathway is operative in a cell-free system of purified and recombinant NADPH oxidase components. Activation by PA + DG was ATP-independent and unaffected by the protein kinase inhibitor staurosporine, indicating the lack of protein kinase involvement. Both PA and DG were required for optimal activation to occur. The drug reduced activation of NADPH oxidase by either arachidonic acid or PA + DG, with IC(50) values of 46 and 25 microm, respectively. The optimal concentration of arachidonic acid or PA + DG for oxidase activation was shifted to the right with, indicating interference of the drug with the interaction of lipid activators and enzyme components. inhibited the lipid-induced aggregation/sedimentation of oxidase components p47(phox) and p67(phox), suggesting a disruption of the lipid-mediated assembly process. The direct effects of on NADPH oxidase activation complicate its use as a "specific" inhibitor of DG kinase. We conclude that the protein kinase independent pathway of NADPH oxidase activation by PA and DG involves direct interaction with NADPH oxidase components. Thus, NADPH oxidase proteins are functional targets for these lipid messengers in the neutrophil. PMID- 11060301 TI - Role of protein kinase Czeta in Ras-mediated transcriptional activation of vascular permeability factor/vascular endothelial growth factor expression. AB - Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF), a multifunctional cytokine, is regulated by different factors including degree of cell differentiation, hypoxia, and certain oncogenes namely, ras and src. The up regulation of VPF/VEGF expression by Ras has been found to be through both transcription and mRNA stability. The present study investigates a novel pathway whereby Ras promotes the transcription of VPF/VEGF by activating protein kinase Czeta (PKCzeta). The Ras-mediated overexpression of VPF/VEGF was also found to be inhibited by using the antisense or the dominant-negative mutant of PKCzeta. In co-transfection assays, by overexpressing oncogenic Ha-Ras (12 V) and PKCzeta, there was an additive effect up to 4-fold in activation of Sp1-mediated VPF/VEGF transcription. It has been shown through electrophoretic mobility shift assay that Ras promoted the PKCzeta-induced binding of Sp1 to the VPF/VEGF promoter. In the presence of PDK-1, a major activating kinase for PKC, the Ras-mediated activation of VPF/VEGF promoter through PKCzeta was further increased, suggesting that PKCzeta can serve as an effector for both Ras and PDK-1. In other experiments, with the use of a dominant-negative mutant of phosphatidylinositol 3 kinase, the activation of VPF/VEGF promoter through Ras, PDK-1, and PKCzeta was completely repressed, indicating phosphatidylinositol 3-kinase as an important component of this pathway. Taken together, these data elucidate the signaling mechanism of Ras-mediated VPF/VEGF transcriptional activation through PKCzeta and also provide insight into PKCzeta and Sp1-dependent transcriptional regulation of VPF/VEGF. PMID- 11060302 TI - The Arabidopsis thaliana isogene NIT4 and its orthologs in tobacco encode beta cyano-L-alanine hydratase/nitrilase. AB - Nitrilases (nitrile aminohydrolases, EC ) are enzymes that catalyze the hydrolysis of nitriles to the corresponding carbon acids. Among the four known nitrilases of Arabidopsis thaliana, the isoform NIT4 is the most divergent one, and homologs of NIT4 are also known from species not belonging to the Brassicaceae like Nicotiana tabacum and Oryza sativa. We expressed A. thaliana NIT4 as hexahistidine tag fusion protein in Escherichia coli. The purified enzyme showed a strong substrate specificity for beta-cyano-l-alanine (Ala(CN)), an intermediate product of cyanide detoxification in higher plants. Interestingly, not only aspartic acid but also asparagine were identified as products of NIT4 catalyzed Ala(CN) hydrolysis. Asn itself was no substrate for NIT4, indicating that it is not an intermediate but one of two reaction products. NIT4 therefore has both nitrilase and nitrile hydratase activity. Several lines of evidence indicate that the catalytic center for both reactions is the same. The NIT4 homologs of N. tabacum were found to catalyze the same reactions and protein extracts of A. thaliana, N. tabacum and Lupinus angustifolius also converted Ala(CN) to Asp and Asn in vitro. NIT4 may play a role in cyanide detoxification during ethylene biosynthesis because extracts from senescent leaves of A. thaliana showed higher Ala(CN) hydratase/nitrilase activities than extracts from nonsenescent tissue. PMID- 11060303 TI - Characterization of the initiation factor eIF2B and its regulation in Drosophila melanogaster. AB - Eukaryotic initiation factor (eIF) 2B catalyzes a key regulatory step in the initiation of mRNA translation. eIF2B is well characterized in mammals and in yeast, although little is known about it in other eukaryotes. eIF2B is a hetropentamer which mediates the exchange of GDP for GTP on eIF2. In mammals and yeast, its activity is regulated by phosphorylation of eIF2alpha. Here we have cloned Drosophila melanogaster cDNAs encoding polypeptides showing substantial similarity to eIF2B subunits from yeast and mammals. They also exhibit the other conserved features of these proteins. D. melanogaster eIF2Balpha confers regulation of eIF2B function in yeast, while eIF2Bepsilon shows guanine nucleotide exchange activity. In common with mammalian eIF2Bepsilon, D. melanogaster eIF2Bepsilon is phosphorylated by glycogen synthase kinase-3 and casein kinase II. Phosphorylation of partially purified D. melanogaster eIF2B by glycogen synthase kinase-3 inhibits its activity. Extracts of D. melanogaster S2 Schneider cells display eIF2B activity, which is inhibited by phosphorylation of eIF2alpha, showing the insect factor is regulated similarly to eIF2B from other species. In S2 cells, serum starvation increases eIF2alpha phosphorylation, which correlates with inhibition of eIF2B, and both effects are reversed by serum treatment. This shows that eIF2alpha phosphorylation and eIF2B activity are under dynamic regulation by serum. eIF2alpha phosphorylation is also increased by endoplasmic reticulum stress in S2 cells. These are the first data concerning the structure, function or control of eIF2B from D. melanogaster. PMID- 11060304 TI - Targeting of membrane proteins to the regulated secretory pathway in anterior pituitary endocrine cells. AB - Unlike the neuroendocrine cell lines widely used to study trafficking of soluble and membrane proteins to secretory granules, the endocrine cells of the anterior pituitary are highly specialized for the production of mature secretory granules. Therefore, we investigated the trafficking of three membrane proteins in primary anterior pituitary endocrine cells. Peptidylglycine alpha-amidating monooxygenase (PAM), an integral membrane protein essential to the production of many bioactive peptides, is cleaved and enters the regulated secretory pathway even when expressed at levels 40-fold higher than endogenous levels. Myc-TMD/CD, a membrane protein lacking the lumenal, catalytic domains of PAM, is still stored in granules. Secretory granules are not the default pathway for all membrane proteins, because Tac accumulates on the surface of pituitary endocrine cells. Overexpression of PAM is accompanied by a diminution in its endoproteolytic cleavage and in its BaCl(2)-stimulated release from mature granules. Because internalized PAM/PAM-antibody complexes are returned to secretory granules, the endocytic machinery of the pituitary endocrine cells is not saturated. As in corticotrope tumor cells, expression of PAM or Myc-TMD/CD alters the organization of the actin cytoskeleton. PAM-mediated alterations in the cytoskeleton may limit maturation of PAM and storage in mature granules. PMID- 11060305 TI - Phage display-selected sequences of the heavy-chain CDR3 loop of the anti-digoxin antibody 26-10 define a high affinity binding site for position 16-substituted analogs of digoxin. AB - The heavy-chain CDR3 region of the high affinity (K(a) = 1.3 x 10(10) M(-)1) anti digoxin monoclonal antibody 26-10 was modified previously to shift its specificity, by substitution of tryptophan 100 by arginine, toward binding analogs of digoxin containing substitutions at position 16. To further change specificity, two 5-mer libraries of the randomly mutagenized phage-displayed 26 10 HCDR3 region (positions 94-98) were panned against digoxin-bovine serum albumin (BSA) as well as against 16-acetylgitoxin-BSA. When a mutant Fab that binds 16-substituted analogs preferentially was used as a parent sequence, clones were obtained with affinities for digoxin increased 2-4-fold, by panning on digoxin-BSA yet retaining the specificity shift. Selection on 16-acetylgitoxin BSA, however, resulted in nine clones that bound gitoxin (16-OH) up to 150-fold higher than the wild-type 26-10, due to a consensus mutation of Ser(H95) to Gly(H95). The residues at both position H95 (serine) and position H100 (tryptophan) contact hapten in the crystal structure of the Fab 26-10-digoxin complex. Thus, by mutating hapten contact residues, it is possible to reorder the combining site of a high affinity antibody, resulting in altered specificity, yet retain or substantially increase the relative affinity for the cross-reactive ligand. PMID- 11060306 TI - Combinatorial control of cyclin B1 nuclear trafficking through phosphorylation at multiple sites. AB - Entry into mitosis is regulated by the Cdc2 kinase complexed to B-type cyclins. We and others recently reported that cyclin B1/Cdc2 complexes, which appear to be constitutively cytoplasmic during interphase, actually shuttle continually into and out of the nucleus, with the rate of nuclear export exceeding the import rate (). At the time of entry into mitosis, the import rate is increased, whereas the export rate is decreased, leading to rapid nuclear accumulation of Cdc2/cyclin B1. Although it has recently been reported that phosphorylation of 4 serines within cyclin B1 promotes the rapid nuclear translocation of Cdc2/cyclin B1 at G(2)/M, the role that individual phosphorylation sites play in this process has not been examined (, ). We report here that phosphorylation of a single serine residue (Ser(113) of Xenopus cyclin B1) abrogates nuclear export of cyclin B1. This serine lies directly within the cyclin B1 nuclear export sequence and, when phosphorylated, prevents binding of the nuclear export factor, CRM1. In contrast, analysis of phosphorylation site mutants suggests that coordinate phosphorylation of all 4 serines (94, 96, 101, and 113) is required for the accelerated nuclear import of cyclin B1/Cdc2 characteristic of G(2)/M. Additionally, binding of cyclin B1 to importin-beta, the factor known to be responsible for the slow interphase nuclear entry of cyclin B1, appears to be unaffected by the phosphorylation state of cyclin B. These data suggest that a distinct import factor must be recruited to enhance nuclear entry of Cdc2/cyclin B1 at the G(2)/M transition. PMID- 11060307 TI - Inhibition of a Gi-activated potassium channel (GIRK1/4) by the Gq-coupled m1 muscarinic acetylcholine receptor. AB - The G protein-coupled inwardly rectifying K+ channel, GIRK1/GIRK4, can be activated by receptors coupled to the Galpha(i) subunit. An opposing role for Galpha(q) receptor signaling in GIRK regulation has only recently begun to be established. We have studied the effects of m1 muscarinic acetylcholine receptor (mAChR) stimulation, which is known to mobilize calcium and activate protein kinase C (PKC) by a Galpha(q)-dependent mechanism, on whole cell GIRK1/4 currents in Xenopus oocytes. We found that stimulation of the m1 mAChR suppresses both basal and dopamine 2 receptor-activated GIRK 1/4 currents. Overexpression of Gbetagamma subunits attenuates this effect, suggesting that increased binding of Gbetagamma to the GIRK channel can effectively compete with the G(q)-mediated inhibitory signal. This G(q) signal requires the use of second messenger molecules; pharmacology implicates a role for PKC and Ca2+ responses as m1 mAChR mediated inhibition of GIRK channels is mimicked by PMA and Ca2+ ionophore. We have analyzed a series of mutant and chimeric channels suggesting that the GIRK4 subunit is capable of responding to G(q) signals and that the resulting current inhibition does not occur via phosphorylation of a canonical PKC site on the channel itself. PMID- 11060308 TI - Glutathiolation of proteins by glutathione disulfide S-oxide derived from S nitrosoglutathione. Modifications of rat brain neurogranin/RC3 and neuromodulin/GAP-43. AB - S-Nitrosoglutathione (GSNO) undergoes spontaneous degradation that generates several nitrogen-containing compounds and oxidized glutathione derivatives. We identified glutathione sulfonic acid, glutathione disulfide S-oxide (GS(O)SG), glutathione disulfide S-dioxide, and GSSG as the major decomposition products of GSNO. Each of these compounds and GSNO were tested for their efficacies to modify rat brain neurogranin/RC3 (Ng) and neuromodulin/GAP-43 (Nm). Among them, GS(O)SG was found to be the most potent in causing glutathiolation of both proteins; four glutathiones were incorporated into the four Cys residues of Ng, and two were incorporated into the two Cys residues of Nm. Ng and Nm are two in vivo substrates of protein kinase C; their phosphorylations by protein kinase C attenuate the binding affinities of both proteins for calmodulin. When compared with their respective unmodified forms, the glutathiolated Ng was a poorer substrate and glutathiolated Nm a better substrate for protein kinase C. Glutathiolation of these two proteins caused no change in their binding affinities for calmodulin. Treatment of [(35)S]cysteine-labeled rat brain slices with xanthine/xanthine oxidase or a combination of xanthine/xanthine oxidase with sodium nitroprusside resulted in an increase in cellular level of GS(O)SG. These treatments, as well as those by other oxidants, all resulted in an increase in thiolation of proteins; among them, thiolation of Ng was positively identified by immunoprecipitation. These results show that GS(O)SG is one of the most potent glutathiolating agents generated upon oxidative stress. PMID- 11060309 TI - SM-20 is a novel mitochondrial protein that causes caspase-dependent cell death in nerve growth factor-dependent neurons. AB - Sympathetic neurons undergo protein synthesis-dependent apoptosis when deprived of nerve growth factor (NGF). Expression of SM-20 is up-regulated in NGF-deprived sympathetic neurons, and ectopic SM-20 is sufficient to promote neuronal death in the presence of NGF. We now report that SM-20 is a mitochondrial protein that promotes cell death through a caspase-dependent mechanism. SM-20 immunofluorescence was present in the cytoplasm in a punctate pattern that colocalized with cytochrome oxidase I and with mitochondria-selective dyes. Analysis of SM-20/dihydrofolate reductase fusion proteins revealed that the first 25 amino acids of SM-20 contain a functional mitochondrial targeting sequence. An amino-terminal truncated form of SM-20 was not restricted to mitochondria but instead localized throughout the cytosol and nucleus. Nevertheless, the truncated SM-20 retained the ability to induce neuronal death, similar to the wild type protein. SM-20-induced death was accompanied by caspase-3 activation and was blocked by a general caspase inhibitor. Additionally, overexpression of SM-20, under conditions where cell death is blocked by a general caspase inhibitor, did not result in widespread release of cytochrome c from mitochondria. These results indicate that SM-20 is a novel mitochondrial protein that may be an important mediator of neurotrophin-withdrawal-mediated cell death. PMID- 11060310 TI - Enoate reductases of Clostridia. Cloning, sequencing, and expression. AB - The enr genes specifying enoate reductases of Clostridium tyrobutyricum and Clostridium thermoaceticum were cloned and sequenced. Sequence comparison shows that enoate reductases are similar to a family of flavoproteins comprising 2,4 dienoyl-coenzyme A reductase from Escherichia coli and old yellow enzyme from yeast. The C. thermoaceticum enr gene product was expressed in recombinant Escherichia coli cells growing under anaerobic conditions. The recombinant enzyme was purified and characterized. PMID- 11060311 TI - KiSS-1 represses 92-kDa type IV collagenase expression by down-regulating NF kappa B binding to the promoter as a consequence of Ikappa Balpha -induced block of p65/p50 nuclear translocation. AB - The 92-kDa type IV collagenase (MMP-9) plays a critical role in tissue remodeling. We undertook a study to determine whether the KiSS-1 gene, previously shown to suppress cancer spread (metastases), negatively regulates MMP-9 expression. Six cell lines positive for MMP-9 mRNA were deficient in KiSS-1 mRNA. One of these cell lines, HT-1080, stably transfected with a KiSS-1 expression construct, demonstrated substantially lower MMP-9 enzyme activity/protein and in vitro invasiveness. The lower MMP-9 enzyme activity reflected reduced steady state mRNA levels which, in turn, was due to attenuated transcription. Activation of ERKs and JNKs by phorbol 12-myristate 13-acetate and tumor necrosis factor alpha, respectively, leading to increased MMP-9 amounts was not antagonized by KiSS-1 expression, suggesting that MAPK pathways modulating MMP-9 synthesis are not the target of KiSS-1. Although MMP-9 expression is regulated by AP-1, Sp1, and Ets transcription factors, KiSS-1 did not alter the binding of these factors to the MMP-9 promoter. However, NF-kappaB binding to the MMP-9 promoter required for expression of this collagenase was reduced by KiSS-1 expression. Diminished NF-kappaB binding reflected less p50/p65 in the nucleus secondary to increased IkappaBalpha levels in the cytosols of the KiSS-1 transfectants. Thus, KiSS-1 diminishes MMP-9 expression by effecting reduced NF-kappaB binding to the promoter. PMID- 11060312 TI - A hydrophobic stretch of 12 amino acid residues in the middle of alpha-synuclein is essential for filament assembly. AB - Neuronal and oligodendrocytic aggregates of fibrillar alpha-synuclein define several diseases of the nervous system. It is likely that these inclusions impair vital metabolic processes and compromise viability of affected cells. Here, we report that a 12-amino acid stretch ((71)VTGVTAVAQKTV(82)) in the middle of the hydrophobic domain of human alpha-synuclein is necessary and sufficient for its fibrillization based on the following observations: 1) human beta-synuclein is highly homologous to alpha-synuclein but lacks these 12 residues, and it does not assemble into filaments in vitro; 2) the rate of alpha-synuclein polymerization in vitro decreases after the introduction of a single charged amino acid within these 12 residues, and a deletion within this region abrogates assembly; 3) this stretch of 12 amino acids appears to form the core of alpha-synuclein filaments, because it is resistant to proteolytic digestion in alpha-synuclein filaments; and 4) synthetic peptides corresponding to this 12-amino acid stretch self polymerize to form filaments, and these peptides promote fibrillization of full length human alpha-synuclein in vitro. Thus, we have identified key sequence elements necessary for the assembly of human alpha-synuclein into filaments, and these elements may be exploited as targets for the design of drugs that inhibit alpha-synuclein fibrillization and might arrest disease progression. PMID- 11060313 TI - MAP-1, a novel proapoptotic protein containing a BH3-like motif that associates with Bax through its Bcl-2 homology domains. AB - A novel Bax-associating protein, named MAP-1 (Modulator of Apoptosis), has been identified in a yeast two-hybrid screen. MAP-1 contains a BH3-like (BH: Bcl-2 homology) motif and mediates caspase-dependent apoptosis in mammalian cells when overexpressed. MAP-1 homodimerizes and associates with the proapoptotic Bax and the prosurvival Bcl-2 and Bcl-X(L) of the Bcl-2 family in vitro and in vivo in mammalian cells. Mutagenesis analyses revealed that the BH3-like domain in MAP-1 is not required for its association with Bcl-X(L) but is required for association with Bax and for mediating apoptosis. Interestingly, in contrast to other Bax associating proteins such as Bcl-X(L) and Bid, which require the BH3 and BH1 domains of Bax, respectively, for binding, the binding of MAP-1 to Bax appears to require all three BH domains (BH1, BH2, and BH3) of Bax, because point mutation of the critical amino acid in any one of these domains is sufficient to abolish its binding to MAP-1. These data suggest that MAP-1 mediates apoptosis through a mechanism that involves binding to Bax. PMID- 11060314 TI - Molecular cloning of a divinyl ether synthase. Identification as a CYP74 cytochrome P-450. AB - Lipoxygenase-derived fatty acid hydroperoxides are metabolized by CYP74 cytochrome P-450s to various oxylipins that play important roles in plant growth and development. Here, we report the characterization of a Lycopersicon esculentum (tomato) cDNA whose predicted amino acid sequence defines a previously unidentified P-450 subfamily (CYP74D). The recombinant protein, expressed in Escherichia coli, displayed spectral properties of a P-450. The enzyme efficiently metabolized 9-hydroperoxy linoleic acid and 9-hydroperoxy linolenic acid but was poorly active against the corresponding 13-hydroperoxides. Incubation of recombinant CYP74D with 9-hydroperoxy linoleic acid and 9 hydroperoxy linolenic acid yielded divinyl ether fatty acids (colneleic acid and colnelenic acid, respectively), which have been implicated as plant anti-fungal toxins. This represents the first identification of a cDNA encoding a divinyl ether synthase and establishment of the enzyme as a CYP74 P-450. Genomic DNA blot analysis revealed the existence of a single divinyl ether synthase gene located on chromosome one of tomato. In tomato seedlings, root tissue was the major site of both divinyl ether synthase mRNA accumulation and enzyme activity. These results indicate that developmental expression of the divinyl ether synthase gene is an important determinant of the tissue specific synthesis of divinyl ether oxylipins. PMID- 11060315 TI - Hypusine is required for a sequence-specific interaction of eukaryotic initiation factor 5A with postsystematic evolution of ligands by exponential enrichment RNA. AB - Hypusine is formed through a spermidine-dependent posttranslational modification of eukaryotic initiation factor 5A (eIF-5A) at a specific lysine residue. The reaction is catalyzed by deoxyhypusine synthase and deoxyhypusine hydroxylase. eIF-5A is the only protein in eukaryotes and archaebacteria known to contain hypusine. Although both eIF-5A and deoxyhypusine synthase are essential genes for cell survival and proliferation, the precise biological function of eIF-5A is unclear. We have previously proposed that eIF-5A may function as a bimodular protein, capable of interacting with protein and nucleic acid (Liu, Y. P., Nemeroff, M., Yan, Y. P., and Chen, K. Y. (1997) Biol. Signals 6, 166-174). Here we used the method of systematic evolution of ligands by exponential enrichment (SELEX) to identify the sequence specificity of the potential eIF-5A RNA targets. The post-SELEX RNA obtained after 16 rounds of selection exhibited a significant increase in binding affinity for eIF-5A with an apparent dissociation constant of 1 x 10(-7) m. The hypusine residue was found to be critical for this sequence specific binding. The post-SELEX RNAs shared a high sequence homology characterized by two conserved motifs, UAACCA and AAUGUCACAC. The consensus sequence was determined as AAAUGUCACAC by sequence alignment and binding studies. BLAST analysis indicated that this sequence was present in > 400 human expressed sequence tag sequences. The C terminus of eIF-5A contains a cold shock domain like structure, similar to that present in cold shock protein A (CspA). However, unlike CspA, the binding of eIF-5A to either the post-SELEX RNA or the 5' untranslated region of CspA mRNA did not affect the sensitivity of these RNAs to ribonucleases. These data suggest that the physiological significance of eIF-5A RNA interaction depends on hypusine and the core motif of the target RNA. PMID- 11060316 TI - THIK-1 and THIK-2, a novel subfamily of tandem pore domain K+ channels. AB - Two cDNAs encoding novel K(+) channels, THIK-1 and THIK-2 (tandem pore domain halothane inhibited K(+) channel), were isolated from rat brain. The proteins of 405 and 430 amino acids were 58% identical to each other. Homology analysis showed that the novel channels form a separate subfamily among tandem pore domain K(+) channels. The genes of the human orthologs were identified as human genomic data base entries. They possess one intron each and were assigned to chromosomal region 14q24.1-14q24.3 (human (h) THIK-1) and 2p22-2p21 (hTHIK-2). In rat (r), THIK-1 (rTHIK-1) is expressed ubiquitously; rTHIK-2 expression was found in several tissues including brain and kidney. In situ hybridization of brain slices showed that rTHIK-2 is strongly expressed in most brain regions, whereas rTHIK-1 expression is more restricted. Heterologous expression of rTHIK-1 in Xenopus oocytes revealed a K(+) channel displaying weak inward rectification in symmetrical K(+) solution. The current was enhanced by arachidonic acid and inhibited by halothane. rTHIK-2 did not functionally express. Confocal microscopy of oocytes injected with green fluorescent protein-tagged rTHIK-1 or rTHIK-2 showed that both channel subunits are targeted to the outer membrane. However, coinjection of rTHIK-2 did not affect the currents induced by rTHIK-1, indicating that the two channel subunits do not form heteromers. PMID- 11060317 TI - Type II transmembrane serine proteases. Insights into an emerging class of cell surface proteolytic enzymes. PMID- 11060318 TI - Introduction PMID- 11060319 TI - Presidential address. PMID- 11060320 TI - Cancer chemoprevention in the 21st century: genetics, risk modeling, and molecular targets. PMID- 11060321 TI - Lynch syndrome: genetics, natural history, genetic counseling, and prevention. AB - Lynch syndrome is the most common hereditary form of colorectal cancer (CRC). Its natural history has been investigated extensively, so that highly targeted surveillance and management strategies, melded to its natural history, have proven effective in cancer control. Most important is the early age of onset of cancer (approximately 44 years), involving CRC and the several extracolonic cancers that are integral to the syndrome. With respect to CRC, approximately 70% of cases occur proximal to the splenic flexure. Synchronous and metachronous CRCs are extremely common. Full colonoscopy should be initiated when the patient is between the ages of 20 and 25, and because of the accelerated carcinogenesis of CRC, it should be performed every 1 to 2 years. The presence of initial CRC requires subtotal colectomy, given the mentioned increased frequency of metachronous cancer. Options available for germ-line mutation carriers, in addition to cancer screening, include prophylactic colectomy as well as prophylactic total abdominal hysterectomy and bilateral salpingo-oophorectomy. The discovery of mismatch repair germ-line mutations (most commonly MSH2 or MLH1) has added significantly to the recognition of this disease as well as to the search for high-risk individuals throughout families who, with genetic counseling, may become candidates for germ-line mutation testing. Clearly, hereditary nonpolyposis colorectal cancer provides an excellent opportunity for learning about the etio-pathogenesis of cancer at the molecular and clinical levels and how this knowledge might ultimately be exploited for cancer control. A search for chemoprevention agents, such as cyclo-oxygenase 2 inhibitors, as well as for putative environmental effects and how they may interact with the genetic component in CRC etiology should abet this entire cancer control process. PMID- 11060322 TI - Postoperative adjuvant chemoradiation therapy for patients with resected gastric cancer: intergroup 116. PMID- 11060323 TI - Chemotherapy for advanced non-small-cell lung cancer: modest progress, many choices. PMID- 11060324 TI - Commentary on "cytoreduction nephrectomy in metastatic renal cancer: the results of Southwest Oncology Group Trial 8949". PMID- 11060325 TI - Notes on the Ontario trial in the context of breast-conserving therapy for early stage breast cancer. PMID- 11060326 TI - Vascular endothelial growth factor as a target for antiangiogenic therapy. PMID- 11060327 TI - Enhanced apoptosis with combination C225/radiation treatment serves as the impetus for clinical investigation in head and neck cancers. AB - PURPOSE: Epidermal growth factor receptor (EGFr) is overexpressed in a majority of head and neck squamous cell carcinomas, and this overexpression is associated with a poor prognosis. Therefore, EGFr has become the target of investigations aimed at disabling the receptor to determine whether this process leads to improved tumor kill with conventional treatment. MATERIALS AND METHODS: C225 is an anti-EGFr monoclonal antibody that inhibits receptor activity by blocking the ligand binding site. A panel of human head and neck squamous cell carcinoma cell lines was used to study the combination of C225 and radiation. RESULTS: It was determined that the combination of C225 (5 microgram/mL) delivered simultaneously with radiation (3 Gy) resulted in a greater decrement in cellular proliferation than either treatment alone. This reduction in proliferation correlated with reduced EGFr tyrosine phosphorylation and a reduction in phosphorylated signal transducer and activator of transcription-3 (STAT-3) protein (known to protect cells from apoptosis). Also, the decrement in proliferation correlated with increased apoptotic events, thereby indirectly linking C225/radiation-induced regulation of STAT-3 protein to apoptosis. CONCLUSION: This preclinical work serves as important support for the ongoing clinical investigation of C225 and radiotherapy for patients with head and neck carcinomas. The initial results of these clinical studies have been promising. PMID- 11060328 TI - New therapeutic agents targeting the epidermal growth factor receptor. PMID- 11060329 TI - The genetic testing process: how much counseling is needed? PMID- 11060330 TI - Organizing cancer genetics programs: the Swiss model. AB - PURPOSE: Inherited predisposition to cancer is a complex issue, and risk assessment, counseling, and management are multidisciplinary tasks. An infrastructure is needed through which clinical services may be delivered and research activities conducted. METHODS: The Swiss Institute for Applied Cancer Research (SIAK) Network for Cancer Predisposition Testing and Counseling was established in 1999. The goals of the Network are to set common standards for clinical services, to coordinate research activities, to address psychosocial and ethical issues concerning genetic counseling and testing, and to translate the knowledge gained into medical practice. RESULTS: An algorithm for genetic testing and counseling of the families studied by the SIAK Network centers has been established in a consensual process. Eight regional centers currently operate according to this algorithm. Furthermore, a proposal has been submitted for the setup of a Swiss databank for the characterization of BRCA1 and BRCA2 mutation patterns and the identification of potentially unique mutations in probands who seek genetic counseling and testing at the respective centers of the SIAK Network. CONCLUSION: Our coordinated effort will ensure state-of-the-art genetic testing and counseling services in Switzerland and will allow us to acquire knowledge for appropriate risk assessment and surveillance/prevention strategies for individuals with an inherited cancer predisposition. PMID- 11060331 TI - Current applications of genetic technology in predisposition testing and microsatellite instability assays. PMID- 11060332 TI - DNA repair defects inactivate tumor suppressor genes and induce hereditary and sporadic colon cancers. PMID- 11060333 TI - Clinical diagnosis and management of hereditary colorectal cancer syndromes. PMID- 11060334 TI - Chemoprevention options for BRCA1 and BRCA2 mutation carriers. AB - BRCA1 and BRCA2 breast cancer predisposition gene mutation carriers are at markedly increased risk of breast and other cancers. The consideration of chemopreventative options will depend on the cancer site and age-specific penetrance curve. Most chemoprevention studies to date have investigated the role of endocrine intervention in women at increased risk of breast cancer, and study results are conflicting. At the present time, there is uncertainty regarding whether endocrine intervention, particularly with tamoxifen, is as effective in BRCA1 and BRCA2 mutation carriers as in other women who are at increased risk of breast cancer because of hormonal factors or genes with moderately conferred cancer risks. Furthermore, if chemoprevention were needed for at least 10 years to produce an effect, new chemoprevention agents will need to be developed for women in their 30s, as the breast cancer risk curves are steepest between 40 and 50 years of age. Consideration is now being given to types of chemoprevention in this younger age group. There is also an increased risk of other cancers (in particular ovarian cancer and, in men, prostate cancer), and considerations regarding chemoprevention will have to encompass cancer at these sites. PMID- 11060335 TI - Prophylactic oophorectomy in BRCA1 and BRCA2 mutation carriers. AB - The availability of genetic testing for inherited mutations in the BRCA1 and BRCA2 genes provides potentially valuable information to women at elevated risk of breast or ovarian cancer. Unfortunately, women who have inherited a mutation in BRCA1 or BRCA2 have relatively few clinical management options available to reduce their risk of developing breast or ovarian cancer. Because most options for ovarian cancer prevention are not highly efficacious, many high-risk women consider the option of bilateral prophylactic oophorectomy (BPO), in the hope that removal of healthy ovarian tissue will reduce their risk of developing invasive malignancy. It is clear that BPO cannot completely prevent the subsequent development of ovarian cancers because reports have been made of patients who have developed cancers of epithelial ovarian origin subsequent to surgery. However, a number of studies have suggested that BPO may reduce the risk of subsequent breast or ovarian cancers in women. In general, these studies have been conducted in women who represent a heterogeneous group with respect to breast/ovarian cancer risk. Only one study of BPO has been undertaken in women whose elevated cancer risk has been based on knowledge of inherited mutations. This study indicated that a 50% to 70% breast cancer risk reduction could be achieved in women with BRCA1 mutations who underwent BPO. However, substantial additional information is required to provide clinically useful information about cancer prevention to women who carry mutations in BRCA1 or BRCA2. PMID- 11060336 TI - Are BRCA1- and BRCA2-associated breast cancers different? PMID- 11060337 TI - Immunophenotypic and pathologic differences between BRCA1 and BRCA2 hereditary breast cancers. AB - Morphologically and clinically, breast cancer is a heterogeneous group of diseases. This heterogeneity may be a manifestation of differences in the molecular genetic events underlying distinct breast cancer pathogenesis pathways. Examination of hereditary breast cancers (HBC), which have in common an underlying germline mutation in BRCA1 or BRCA2, may provide further insight into this concept. Multiple studies have confirmed that BRCA1-associated HBC (BRCA1 HBC) generally exhibit a specific phenotype that is characterized by high tumor grade and estrogen receptor negativity. Conversely, discrepancies exist between the findings of studies that have examined BRCA2-HBC, and a specific phenotype has not been consistently described. The characteristic phenotype of BRCA1 associated tumors may prove a useful additional tool in selecting individuals with breast cancer who should be offered BRCA1 mutation testing, although further studies are required. Lastly, evidence is emerging to suggest that BRCA1 might be involved in the pathogenesis of a subgroup of non-HBC (by gene underexpression rather than mutation) and that these tumors may exhibit the same phenotype as their hereditary counterparts. PMID- 11060338 TI - Are BRCA1- and BRCA2-associated breast cancers different? Prognosis of BRCA1 associated breast cancer. AB - PURPOSE: To review the available literature regarding the outcome of breast cancer arising in the setting of a germline BRCA1 mutation. METHODS: Reports of relevant studies obtained from a search of MEDLINE and studies referenced in those reports were reviewed. In addition, data from a previously published study of breast conservation therapy among women of Ashkenazi descent were reanalyzed to discern the effect of germline BRCA1 status. RESULTS: Although BRCA1 associated breast cancers have been associated repeatedly with poor prognostic features, studies that examine the impact of germline BRCA1 status on outcome have not consistently shown an adverse effect of BRCA1 mutations. In part, this may result from methodologic limitations of earlier trials. Recent studies that examine relatively unselected groups of Ashkenazi women have indicated a negative prognostic effect of specific BRCA1 mutations. It remains to be determined, however, whether BRCA1 status is an independent prognostic factor and whether the findings can be generalized to other mutations. CONCLUSION: BRCA1-associated breast cancers present with histopathologic features that seem to confer an adverse prognosis, at least in certain subgroups. However, it is not clear whether BRCA1 status has an independent effect on outcome. Until further data become available, treatment for BRCA1-associated breast cancer should be determined by the same factors that influence therapy of nonhereditary disease. PMID- 11060339 TI - Prognostic significance of germline BRCA2 mutations in hereditary breast cancer patients. AB - PURPOSE: Breast cancer in BRCA2 gene mutation carriers differs from BRCA1 associated breast cancer or so-called sporadic breast cancer in clinical features and behavior. These differences may be of importance for the prevention, screening, and ultimately treatment of breast cancer in women with such germline mutations. METHODS: We reviewed the few studies that have reported on survival in patients with BRCA2-associated breast cancer. In this article we discuss why family history is no substitute for hereditary breast cancer with regard to studying survival and possible reasons why studies using family history yield contradictory results, why BRCA2-associated breast cancer should be considered a unique entity, and what methodological problems may exist, especially with regard to family-based studies. RESULTS: Five studies have reported on survival in BRCA2 associated breast cancer. Two studies showed a statistically significant worse survival for BRCA2 patients, but the patients from one of these studies were later claimed to have a trend toward better prognosis when controls were matched for age and year of diagnosis. The other study found that the unfavorable prognosis of BRCA2 patients was, to a great extent, due to a worse stage of the disease at time of diagnosis. The remaining three studies showed no significant effect of germline BRCA2 mutations on survival. The numbers of BRCA2 patients investigated in these studies were 42, 20, 23, 28, and 54 patients. Five-year overall survival in these patients varied from 65% to 74%. CONCLUSION: No definite conclusion can be made with regard to the prognosis of BRCA2-associated breast cancer, but large differences in comparison with sporadic breast cancer are not likely to exist. Breast cancer caused by BRCA2 mutations is also a distinct entity with its own features when compared with BRCA1-associated breast cancer. In contrast with BRCA1-associated breast cancer, BRCA2 tumors tend to be more often steroid receptor-positive. PMID- 11060340 TI - Aggregation, fusion, and vesicle formation of modified low density lipoprotein particles: molecular mechanisms and effects on matrix interactions. AB - Initiation of atherosclerosis is characterized by accumulation of aggregates of small lipid droplets and vesicles in the extracellular matrix of the arterial intima. The droplets and vesicles have features that suggest that they are formed from modified plasma-derived low density lipoprotein (LDL) particles. A variety of hydrolytic enzymes and prooxidative agents that could lead to extracellular assembly of LDL-derived droplets and vesicles are present in the arterial intima. In fact, in vitro studies have demonstrated that extensive oxidation of LDL and treatment of LDL with either proteolytic or lipolytic enzymes will induce LDL aggregation and fusion and treatment of LDL with cholesterol esterase will cause formation of vesicles. Fusion of LDL particles proceeds faster in vitro when they are bound to components of the extracellular matrix derived from the arterial intima, such as proteoglycans, and, depending on the type of modification, the strength of binding of modified LDL to the matrix components may either increase or decrease. In the present article, we discuss molecular mechanisms that provide clues as to how aggregated lipid droplets and vesicles may be derived from modified LDL particles. We also describe how these modified forms of LDL, by means of their trapping to the extracellular matrix, may lead to extracellular lipid accumulation in the arterial intima. PMID- 11060341 TI - Rapid initial removal of chylomicron remnants by the mouse liver does not require hepatically localized apolipoprotein E. AB - Apolipoprotein E (apoE) is a ligand for the low density lipoprotein receptor (LDLR) and the low density lipoprotein receptor-related protein (LRP). The aim of the present study was to clarify the role of hepatically localized apoE in the rapid initial removal of chylomicron remnants by using the isolated perfused liver. Radiolabeled chylomicron remnants were perfused in a single nonrecirculating pass into the livers of C57BL/6J (wild-type) mice, apoE-knockout mice, and apoE/LDLR-knockout mice for a period of 20 min. Aliquots of the perfusate leaving the liver were collected at regular intervals and the rate of removal of radioactivity was determined. At a trace concentration of chylomicron remnants (0.05 microgram of protein per ml), wild-type mouse livers removed at a steady state of 50-55% of total chylomicron remnants perfused per pass; livers from apoE-knockout mice had the same capacity as wild-type mouse livers. When the concentration of remnants was increased to 12 microgram of protein per ml, a level at which it has been shown that LDL receptor and LRP are near saturation, the capacity of the wild-type mouse livers to remove chylomicron remnants was decreased to 10-25% per pass, confirming that the removal mechanisms were nearing saturation. However, instead of finding a greater reduction in the removal rates or impairment in chylomicron remnant removal, livers from apoE-knockout mice were just as efficient as those from wild-type mice in removing remnants. Livers of mice that lacked both apoE and the LDLR also had a similar rate of removal at relatively low remnant concentrations (0.05-0.5 microgram/ml), but had reduced capacity in removing remnants at a relatively high concentration (4-12 microgram/ml) of chylomicron remnants ( approximately 20% per pass). The rate of removal at these concentrations, however, was similar to that attributed to the LRP in previous studies. Chylomicron remnants, whose apolipoproteins were disrupted by trypsinization, were removed at a normal rate by wild-type mouse livers but there was almost no removal by apoE-knockout mouse livers. At higher concentrations, however, the removal of apolipoprotein-disrupted chylomicron remnants was decreased. Our present findings do not support the hypothesis that hepatically localized apoE is a critical factor in the rapid initial removal of chylomicron remnants by either of the major pathways but do suggest that hepatically localized apoE can be added to lipoproteins to accelerate their uptake, although this process may have a limited capacity to compensate for apoE deficiency on lipoproteins. PMID- 11060342 TI - Intracellular events in the assembly of chylomicrons in rabbit enterocytes. AB - The aim of this study was to determine the intracellular events in chylomicron assembly in adult villus enterocytes. We have used novel methods for separation of the intracellular components of the secretory compartment [rough and smooth endoplasmic reticulum (RER and SER, respectively) and Golgi], and their membrane and luminal components, from villus enterocytes isolated from rabbit small intestine. The steady state composition of the components of the secretory compartment and the intracellular pools of newly synthesized apolipoprotein B-48 (apoB-48) and triacylglycerol (TAG) was determined. The observations indicate that the SER is the main site of TAG synthesis and of chylomicron assembly. Newly synthesized apoB-48 and TAG accumulate in the SER membrane and are transferred into the lumen in a microsomal triglyceride transfer protein-dependent step. In enterocytes isolated from chow-fed rabbits, in which fat absorption is relatively slow, transfer of apoB-48 and TAG from the SER membrane into the lumen appears to be rate limiting. In enterocytes from fat-fed rabbits, TAG accumulates in the lumen of the SER, suggesting that movement out of the SER lumen becomes rate limiting, when chylomicron secretion is markedly stimulated. In these cells, the cytosolic TAG also increased to 450 microgram/g enterocytes, compared with 12 microgram/g enterocytes from chow-fed rabbits, indicating that transfer of TAG from the SER membrane into the secretory pathway can become saturated, so that newly synthesized TAG moves into the cytosol. PMID- 11060343 TI - Lipid-binding proteins modulate ligand-dependent trans-activation by peroxisome proliferator-activated receptors and localize to the nucleus as well as the cytoplasm. AB - Peroxisome proliferator-activated receptors (PPARs) are activated by a variety of fatty acids, eicosanoids, and hypolipidemic and insulin-sensitizing drugs. Many of these compounds bind avidly to members of a family of small lipid-binding proteins, the fatty acid-binding proteins (FABPs). Fatty acids are activated to CoA esters, which bind with high affinity to the acyl-CoA-binding protein (ACBP). Thus, the availability of known and potential PPAR ligands may be regulated by lipid-binding proteins. In this report we show by transient transfection of CV-1 cells that coexpression of ACBP and adipocyte lipid-binding protein (ALBP) exerts a ligand- and PPAR subtype-specific attenuation of PPAR-mediated trans activation, suggesting that lipid-binding proteins, when expressed at high levels, may function as negative regulators of PPAR activation by certain ligands. Expression of ACBP, ALBP, and keratinocyte lipid-binding protein (KLBP) is induced during adipocyte differentiation, a process during which PPARgamma plays a prominent role. We present evidence that endogenous ACBP, ALBP, and KLBP not only localize to the cytoplasm but also exhibit a prominent nuclear localization in 3T3-L1 adipocytes. In addition, forced expression of ACBP, ALBP, and KLBP in CV-1 cells resulted in a substantial accumulation of all three proteins in the nucleus. These results suggest that lipid-binding proteins, contrary to the general assumption, may exert their action in the nucleus as well as in the cytoplasm. PMID- 11060344 TI - Mitochondrial and peroxisomal targeting of 2-methylacyl-CoA racemase in humans. AB - 2-Methylacyl-CoA racemase is an auxiliary enzyme required for the peroxisomal beta-oxidative breakdown of (2R)-pristanic acid and the (25R)-isomer of C(27) bile acid intermediates. The enzyme activity is found not only in peroxisomes but also is present in mitochondria of human liver and fibroblasts. The C terminus of the human racemase, a protein of 382 amino acids with a molecular mass of 43,304 daltons as deduced from its cloned cDNA, consists of KASL. Hitherto this sequence has not been recognized as a peroxisomal targeting signal (PTS1). From the in vitro interaction between recombinant racemase and recombinant human PTS1 receptor (Pex5p), and the peroxisomal localization of green fluorescent protein (GFP) fused to the N terminus of full-length racemase or its last six amino acids in tranfected Chinese hamster ovary (CHO) cells, we concluded that ASL is a new PTS1 variant. To be recognized by Pex5p, however, the preceding lysine residue is critical. As shown in another series of transfection experiments with GFP fused to the C terminus of the full-length racemase or racemase with deletions of the N terminus, mitochondrial targeting information is localized between amino acids 22 and 85.Hence, our data show that a single transcript gives rise to a racemase protein containing two topogenic signals, explaining the dual cellular localization of the activity. PMID- 11060345 TI - Characterization of the lipid-binding properties and lipoprotein lipase inhibition of a novel apolipoprotein C-III variant Ala23Thr. AB - We have identified a G-to-A transition in exon 3 of the APOC3 gene resulting in a novel Ala23Thr apolipoprotein (apo) C-III variant, associated with apoC-III deficiency in three unrelated Yucatan Indians. The Ala23Thr substitution modifies the hydrophobic/hydrophilic repartition of the helical N-terminal peptide and hence could disturb the lipid association. In vitro expression in Escherichia coli of wild-type and mutant apoC-III enabled the characterization of the variant. Compared with wild-type apoC-III-Ala23, the mutant apoC-III-Thr23 showed reduced affinity for dimyristoylphosphatidylcholine (DMPC) multilamellar vesicles with higher amounts of free apoC-III. Displacement of apoE from discoidal apoE:dipalmitoylphosphatidycholine (DPPC) complex by apoC-III-Thr23 was comparable to wild type but the less efficient binding of the apoC-III-Thr23 to the discoidal complex resulted in a higher apoE/apoC-III (mol/mol) ratio (34%) than with wild-type/apoE:DPPC mixtures. The inhibition of lipoprotein lipase (LPL) by apoC-III-Thr23 was comparable to that of wild type, and therefore effects on LPL activity could not explain the lower triglyceride (Tg) levels in Thr-23 carriers. Thus, these in vitro results suggest that in vivo the less efficient lipid binding of apoC-III-Thr23 might lead to a faster catabolism of free apoC-III, reflected in the reduced plasma apoC-III levels identified in Thr 23 carriers, and poorer competition with apoE, which might enhance clearance of Tg-rich lipoproteins and lower plasma Tg levels seen in Thr-23 carriers. PMID- 11060346 TI - Aberrant pathways in the late stages of cholesterol biosynthesis in the rat. Origin and metabolic fate of unsaturated sterols relevant to the Smith-Lemli Opitz syndrome. AB - Minor aberrant pathways of cholesterol biosynthesis normally produce only trace levels of abnormal sterol metabolites but may assume major importance when an essential biosynthetic step is blocked. Cholesta-5,8-dien-3beta-ol, its Delta(5,7) isomer, and other noncholesterol sterols accumulate in subjects with the Smith-Lemli-Opitz syndrome (SLOS), a severe developmental disorder caused by a defective Delta(7) sterol reductase gene. We have explored the formation and metabolism of unsaturated sterols relevant to SLOS by incubating tritium-labeled Delta(5,8), Delta(6, 8), Delta(6,8(14)), Delta(5,8(14)), and Delta(8) sterols with rat liver preparations. More than 60 different incubations were carried out with washed microsomes or the 10,000 g supernatant under aerobic or anaerobic conditions; some experiments included addition of cofactors, fenpropimorph (a Delta(8);-Delta(7) isomerase inhibitor), and/or AY-9944 (a Delta(7) reductase inhibitor). The tritium-labeled metabolites from each incubation were identified by silver ion high performance liquid chromatography on the basis of their coelution with unlabeled authentic standards, as free sterols and/or acetate derivatives. The Delta(5,8) sterol was converted slowly to cholesterol via the Delta(5,7) sterol, which also slowly isomerized back to the Delta(5,8) sterol. The Delta(6,8) sterol was metabolized rapidly to cholesterol by an oxygen requiring pathway via the Delta(7,9(11)), Delta(8), Delta(7), and Delta(5,7) sterols as well as by an oxygen-independent route involving initial isomerization to the Delta(5,7) sterol. The Delta(8) sterol was partially metabolized to Delta(5,8), Delta(6,8), Delta(7,9(11)), and Delta(5,7,9(11)) sterols when isomerization to Delta(7) was blocked.The combined results were used to formulate a scheme of normal and aberrant biosynthetic pathways that illuminate the origin and metabolic fate of abnormal sterols observed in SLOS and chondrodysplasia punctata. PMID- 11060347 TI - Human apolipoprotein E7:lysine mutations in the carboxy-terminal domain are directly responsible for preferential binding to very low density lipoproteins. AB - Apolipoprotein E7 (apoE7) (apoE3 E244K/E245K) is a naturally occurring mutant in humans that is associated with increased plasma lipid levels and accelerated atherosclerosis. It is reported to display defective binding to low density lipoprotein (LDL) receptors, high affinity binding for heparin, and like apoE4, preferential association with very low density lipoproteins (VLDL). There are two potential explanations for the preference of apoE7 for VLDL: lysine mutations, which occur in the major lipid-binding region (residues 244-272) of the carboxy terminal domain of apoE7, could either directly determine the lipoprotein-binding preference or could interact with negatively charged residues in the amino terminal domain, resulting in a domain interaction similar to that in apoE4 (interaction of Arg-61 with Glu-255), which is responsible for the apoE4 VLDL preference. To distinguish between these possibilities, we determined the binding preferences of recombinant apoE7 and two amino-terminal domain mutants, apoE7 (E49Q/E50Q) and apoE7 (D65N/E66Q), to VLDL-like emulsion particles. ApoE7 and both mutants displayed a higher preference for the emulsion particles than did apoE3, indicating that the carboxy-terminal lysine mutations in apoE7 are directly responsible for its preference for VLDL. Supporting this conclusion, the carboxy-terminal domain 12-kDa fragment of apoE7 (residues 192;-299) displayed a higher preference for VLDL emulsions than did the wild-type fragment. In addition, lipid-free apoE7 had a higher affinity for heparin than did apoE. However, when apoE7 was complexed with dimyristoylphosphatidylcholine or VLDL emulsions, the affinity difference was eliminated. In contrast to previous studies, we found that apoE7 does not bind defectively to the LDL receptor, as determined in both cell culture and solid-phase assays. We conclude that the two additional lysine residues in the carboxy-terminal domain of apoE7 directly alter its lipid- and heparin-binding affinities. These characteristics of apoE7 could contribute to its association with increased plasma lipid levels and atherosclerosis. PMID- 11060348 TI - Long-term kinetic study of beta-carotene, using accelerator mass spectrometry in an adult volunteer. AB - We present a sensitive tracer method, suitable for in vivo human research, that uses beta-[(14)C]carotene coupled with accelerator mass spectrometry (AMS) detection. Using this approach, the concentration-time course of a physiological (306 microgram 200 nCi) oral dose of beta-[(14)C]carotene was determined for 209 days in plasma. Analytes included beta-[(14)C]carotene, [(14)C]retinyl esters, [(14)C]retinol, and several [(14)C]retinoic acids. There was a 5.5-h lag between dosing and the appearance of (14)C in plasma. Labeled beta-carotene and [(14)C]retinyl esters rose and displayed several maxima with virtually identical kinetic profiles over the first 24-h period; elevated [(14)C]retinyl ester concentrations were sustained in the plasma compartment for >21 h postdosing. The appearance of [(14)C]retinol in plasma was also delayed 5.5 h postdosing and its concentration rose linearly for 28 h before declining. Cumulative urine and stool were collected for 17 and 10 days, respectively, and 57.4% of the dose was recovered in the stool within 48 h postdosing. The stool was the major excretion route for the absorbed dose. The turnover times (1/k(el)) for beta-carotene and retinol were 58 and 302 days, respectively. Area under the curve analysis of the plasma response curves suggested a molar vitamin A value of 0.53 for beta carotene, with a minimum of 62% of the absorbed beta-carotene being cleaved to vitamin A.In summary, AMS is an excellent tool for defining the in vivo metabolic behavior of beta-carotene and related compounds at physiological concentrations. Further, our data suggest that retinyl esters derived from beta-carotene may undergo hepatic resecretion with VLDL in a process similar to that observed for beta-carotene. PMID- 11060349 TI - Pristanic acid and phytanic acid: naturally occurring ligands for the nuclear receptor peroxisome proliferator-activated receptor alpha. AB - Phytanic acid and pristanic acid are branched-chain fatty acids, present at micromolar concentrations in the plasma of healthy individuals. Here we show that both phytanic acid and pristanic acid activate the peroxisome proliferator activated receptor alpha (PPARalpha) in a concentration-dependent manner. Activation is observed via the ligand-binding domain of PPARalpha as well as via a PPAR response element (PPRE). Via the PPRE significant induction is found with both phytanic acid and pristanic acid at concentrations of 3 and 1 microM, respectively. The trans-activation of PPARdelta and PPARgamma by these two ligands is negligible. Besides PPARalpha, phytanic acid also trans-activates all three retinoic X receptor subtypes in a concentration-dependent manner. In primary human fibroblasts, deficient in phytanic acid alpha-oxidation, trans activation through PPARalpha by phytanic acid is observed. This clearly demonstrates that phytanic acid itself, and not only its metabolite, pristanic acid, is a true physiological ligand for PPARalpha. Because induction of PPARalpha occurs at ligand concentrations comparable to the levels found for phytanic acid and pristanic acid in human plasma, these fatty acids should be seen as naturally occurring ligands for PPARalpha. These results demonstrate that both pristanic acid and phytanic acid are naturally occurring ligands for PPARalpha, which are present at physiological concentrations. PMID- 11060350 TI - Specific linoleate deficiency in the rat does not prevent substantial carbon recycling from [(14)C]linoleate into sterols. AB - Compared with classic essential fatty acid deficiency or the feeding of a fat free diet, little is known about specific linoleate deficiency in the rat. Carbon recycling into de novo lipogenesis has been reported to be an obligatory feature of linoleate metabolism in the liver, even in extreme linoleate deficiency (LA D). The present study had two objectives: 1) to report a brief summary of the tissue n-6 polyunsaturated fatty acid (PUFA) profiles in specific LA-D, and 2) to quantify whole body carbon recycling from [(14)C]linoleate in specific LA-D. Rats consumed a linoleate-deficient diet for 12 weeks and then received a bolus of [1 (14)C]linoleate by gavage. In linoleate-deficient rats, the triene/tetraene ratio in several organs increased by 18- to 100-fold. The amount of (14)C appearing in organ sterols (dpm/g) of linoleate-deficient rats was 2- to 10-fold higher than in the controls and equaled 16.3% of the [(14)C]linoleate dose given, compared with 7.4% in the controls. We conclude that a similar amount (about 10%) of the carbon skeleton of linoleate is normally recycled into lipids synthesized de novo, as remains in the whole body pool of n-6 polyunsaturates. PMID- 11060351 TI - Context-dependent and invariant associations between lipids, lipoproteins, and apolipoproteins and apolipoprotein E genotype. AB - Variation in apolipoprotein (apo)E genotypes predicts variation in plasma cholesterol and apoB; however, the context-dependent associations between high density lipoprotein (HDL) cholesterol, apoA-I, triglycerides, and lipoprotein[a] (Lp[a]) and this polymorphism remain unsettled. We genotyped 5,025 women and 4,035 men sampled to represent a white general population in the age range 20 to 80+ years (mean ages 58 and 57 years for women and men, respectively). The relative frequencies of the varepsilon22, varepsilon32, varepsilon42, varepsilon33, varepsilon43, and varepsilon44 genotypes were 0.005, 0.127, 0.027, 0.564, 0.251, and 0. 027, respectively. Variations in apoE genotype (in the order listed above) predicted stepwise increases in cholesterol and apoB in both genders (all ANOVAs: P < 0.001), and stepwise decreases in HDL cholesterol and apoA-I in women (both ANOVAs: P < 0.001), but not in men. In both genders varepsilon33 individuals had the lowest levels of nonfasting triglycerides, whereas the highest levels were found in individuals with varepsilon22 and varepsilon44 genotypes (both ANOVAs: P < 0.001). Finally, a stepwise increase in Lp[a] was seen in women (ANOVA: P < 0.001), but not in men. In women, the association between variation in nonfasting triglycerides and Lp[a], and variation in apoE genotypes was mainly seen in those with the highest alcohol consumption, similar to the consumption of most men. Variations in apoE genotype predicted 5% and 11% in women, and 2% and 6% in men, of the total variation in plasma cholesterol and apoB, respectively. Variation in levels of plasma lipoproteins is associated with variation in apoE genotypes in the population at large, with the most pronounced association in women, except for nonfasting triglycerides, for which the association is most pronounced in men.Whereas the associations between variation in plasma cholesterol and apoB and the variation in apoE genotypes seem invariant, the associations with variation in plasma HDL cholesterol, apoA-I, nonfasting triglycerides, and Lp[a] seem context dependent. PMID- 11060352 TI - Intracellular trafficking of pigeon beta-very low density lipoprotein and low density lipoprotein at low and high concentrations in pigeon macrophages. AB - Foam cell formation occurs in vitro at lipoprotein concentrations above 50 microgram/ml in pigeon macrophages. Hypothetically, intracellular trafficking of lipoproteins at higher concentrations may differ from uptake of lipoproteins associated with low concentrations, revealing a separate atherogenic endocytic pathway. Macrophage intracellular trafficking of pigeon beta-very low density lipoprotein (beta-VLDL) and low density lipoprotein (LDL) at low concentrations (12 microgram/ml) near the saturation of high affinity binding sites and high lipoprotein concentrations (50-150 microgram/ml) used to induce foam cell formation were examined. Pigeon beta-VLDL and LDL, differentially labeled with colloidal gold, were added simultaneously to contrast trafficking of beta-VLDL, which causes in vitro foam cell formation, with LDL, which does not. The binding of lipoproteins to cell surface structures, distribution of lipoproteins in endocytic organelles, and the extent of colabeling in the endocytic organelles were determined by thin-section transmission electron microscopy. At low concentrations, the intracellular trafficking of pigeon LDL and beta-VLDL was identical. At high concentrations, LDL was removed more rapidly from the plasma membrane and reached lysosomes more quickly than beta-VLDL. No separate endocytic route was present at high concentrations of beta-VLDL; rather, an increased residence on the plasma membrane, association with nonmicrovillar portions of the plasma membrane, and slower trafficking in organelles of coated-pit endocytosis reflected a more atherogenic trafficking pattern. PMID- 11060353 TI - Kinetic steps for the hydrolysis of sphingomyelin by Bacillus cereus sphingomyelinase in lipid monolayers. AB - The sphingomyelinase (Sphmase) activity degrading sphingomyelin (Sphm) monolayers shows a slow-reaction latency period before exhibiting constant rate catalysis. These two kinetic regions are regulated independently by the lateral surface pressure and by lipids that are biomodulators of cell function such as ceramide, glycosphingolipids, fatty acids, and lysophospholipids. Knowledge of the interfacial adsorption of Sphmase, precatalytic activation, initiation of effective catalysis, and the corresponding kinetic parameters is necessary for studying the level at which different lipids modulate the activity. We dissected some kinetic steps and determined the rate constants for degradation of Sphm, under controlled intermolecular organization, by Sphmase. Six models, adapted to two dimensions, were used to elucidate possible mechanisms for the interfacial activation of Sphmase during the lag time. The models consider enzyme binding to the substrate monolayer and a subsequent, essentially irreversible interfacial activation; this is supported experimentally by monolayer transfer experiments. Some mechanisms involve enzyme-substrate binding and associated states of the enzyme in the bulk subphase or at the interface, prior to complete activation. The activity of Sphmase is consistent with kinetics involving enzyme partitioning into the interface followed by substrate association, and by a process that proceeds with bimolecular kinetic dependence on the interfacial Sphmase concentration, and a subsequent slow step of activation. A possible equilibrium between the apparent monomolecular and bimolecular activated states of the interfacial enzyme, coupled to a slow activation, constitute rate-limiting steps that can explain the existence of lag time and the achievement of a maximum constant rate of degradation of Sphm monolayers by Sphmase. PMID- 11060354 TI - Functional expression of a high affinity mammalian hepatic choline/organic cation transporter. AB - Uptake by the liver of the organic cation and essential nutrient choline is required for the hepatic synthesis of phosphatidylcholine. Uptake of other organic cations is also important for the metabolism and secretion of numerous endobiotics and drugs. Although a high affinity mammalian hepatic choline transporter has been kinetically defined, it has not been previously identified. We have developed stable transfectants of BALB/3T3 cells, using a murine member of the organic cation transporter gene family (mOct1/Slc22a1), and used these cells to characterize the transport of the organic cation choline and model organic cation tetraethylammonium (TEA). Functional expression of mOct1/Slc22a1 in BALB/3T3 cells confers the saturable, temperature-dependent uptake of choline with a K(m) of 42 micrometer, and uptake of TEA with a K(m) of 43 micrometer. We subsequently used our cell culture uptake system to kinetically define in HepG2 cells a high affinity choline uptake process, which transports choline with a K(m) similar to that of mOct1/Slc22a1 protein. We also demonstrated that organic cation transport by mOct1/Slc22a1 is inhibited by several organic cations, and that the gene is expressed in the perinatal period, at a time when phosphatidylcholine synthesis increases. We conclude that mOct1/Slc22a1 encodes a high affinity mammalian hepatic choline/organic cation transporter. This transporter may be important for hepatic phosphatidylcholine synthesis, and for the metabolism and secretion of many organic cationic drugs. PMID- 11060355 TI - HDL modification by secretory phospholipase A(2) promotes scavenger receptor class B type I interaction and accelerates HDL catabolism. AB - During inflammatory states plasma levels of high density lipoprotein (HDL) cholesterol and apolipoprotein A-I (apoA-I) are reduced. Secretory group IIa phospholipase A(2) (sPLA(2)) is a cytokine-induced acute-phase enzyme associated with HDL. Transgenic mice overexpressing sPLA(2) have reduced HDL levels. Studies were performed to define the mechanism for the HDL reduction in these mice. HDL isolated from sPLA(2) transgenic mice have a significantly lower phospholipid content and greater triglyceride content. In autologous clearance studies, (125)I labeled HDL from sPLA(2) transgenic mice was catabolized significantly faster than HDL from control mice (4.24 +/- 1.16 vs. 2.84 +/- 0.1 pools per day, P < 0.008). In both sPLA(2) transgenic and control mice, the cholesteryl ester component of HDL was more rapidly catabolized than the protein component, indicating a selective uptake mechanism. In vitro studies using CHO cells transfected with scavenger receptor class B type I (SR-BI) showed that sPLA(2) modified HDL was nearly twice as efficient as a substrate for cholesteryl ester transfer. These data were confirmed in in vivo selective uptake experiments using adenoviral vector overexpression of SR-BI. In these studies, increased hepatic selective uptake was associated with increased (125)I-labeled apolipoprotein uptake in the kidney. We conclude that during inflammation sPLA(2) hydrolysis of HDL phospholipids alters the lipid composition of the particle, allowing for more efficient SR-BI-mediated selective cholesteryl ester uptake. This enhanced SR-BI activity generates HDL remnants that are preferentially catabolized in the kidney. PMID- 11060356 TI - Lipoprotein lipase- and hepatic triglyceride lipase- promoted very low density lipoprotein degradation proceeds via an apolipoprotein E-dependent mechanism. AB - Apolipoprotein E (apoE) is the primary recognition signal on triglyceride-rich lipoproteins responsible for interacting with low density lipoprotein (LDL) receptors and LDL receptor-related protein (LRP). It has been shown that lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) promote receptor mediated uptake and degradation of very low density lipoproteins (VLDL) and remnant particles, possibly by directly binding to lipoprotein receptors. In this study we have investigated the requirement for apoE in lipase-stimulated VLDL degradation. We compared binding and degradation of normal and apoE-depleted human VLDL and apoE knockout mouse VLDL in human foreskin fibroblasts. Surface binding at 37 degrees C of apoE knockout VLDL was greater than that of normal VLDL by 3- and 40-fold, respectively, in the presence of LPL and HTGL. In spite of the greater stimulation of surface binding, lipase-stimulated degradation of apoE knockout mouse VLDL was significantly lower than that of normal VLDL (30, 30, and 80%, respectively, for control, LPL, and HTGL treatments). In the presence of LPL and HTGL, surface binding of apoE-depleted human VLDL was, respectively, 40 and 200% of normal VLDL whereas degradation was, respectively, 25 and 50% of normal VLDL. LPL and HTGL stimulated degradation of normal VLDL in a dose-dependent manner and by a LDL receptor-mediated pathway. Maximum stimulation (4-fold) was seen in the presence LPL (1 microgram/ml) or HTGL (3 microgram/ml) in lovastatin-treated cells. On the other hand, degradation of apoE depleted VLDL was not significantly increased by the presence of lipases even in lovastatin-treated cells. Surface binding of apoE-depleted VLDL to metabolically inactive cells at 4 degrees C was higher in control and HTGL-treated cells, but unchanged in the presence of LPL. Degradation of prebound apoE-depleted VLDL was only 35% as efficient as that of normal VLDL. Surface binding of apoE knockout or apoE-depleted VLDL was to heparin sulfate proteoglycans because it was completely abolished by heparinase treatment. However, apoE appears to be a primary determinant for receptor-mediated VLDL degradation. Our studies suggest that overexpression of LPL or HTGL may not protect against lipoprotein accumulation seen in apoE deficiency. PMID- 11060357 TI - Deletion of the propeptide of apolipoprotein A-I reduces protein expression but stimulates effective conversion of prebeta-high density lipoprotein to alpha-high density lipoprotein. AB - The properties of the mature and pro-forms of recombinant apolipoprotein A-I (apoA-I) were compared with those of apoA-I isolated from human plasma. When the synthesis and secretion of pro- and mature forms of apoA-I from a baculovirus/insect cell expression system were compared in parallel experiments, the amount of the pro-form of apoA-I synthesized and secreted was severalfold higher than that of the mature form of apoA-I. A comparison of the properties of the pro- and mature forms of recombinant apoA-I and human plasma apoA-I showed no difference between all three in their secondary structure, their ability to self associate, lipid-binding capacity, lecithin: cholesterol acyltransferase activation, and binding to the phospholipid transfer protein. The properties of reconstituted high density lipoprotein (HDL) particles formed from the proteins and their ability to promote cholesterol and phospholipid efflux from human skin fibroblasts were also similar. However, their ability to bind to plasma HDL subfractions differed, because twice as much proapoA-I associated with prebeta(1) HDL and prebeta(2)-HDL subfractions compared with both mature recombinant and plasma apoA-I. Correspondingly, the amount of proapoA-I in alpha-HDL subfractions, especially in alpha(1)-HDL and alpha(2)-HDL, was decreased. We conclude that while the propeptide of apoA-I is required for the effective synthesis and secretion of apoA-I, cleavage of this peptide is a requisite for the effective interconversion of HDL subfractions. PMID- 11060358 TI - Hyperabsorption and retention of campestanol in a sitosterolemic homozygote: comparison with her mother and three control subjects. AB - We measured the percent absorption, turnover, and distribution of campestanol (24 methyl-5alpha-cholestan-3beta-ol) in a sitosterolemic homozygote, her obligate heterozygous mother, and three healthy human control subjects. For reasons relating to sterol hyperabsorption, the homozygote consumed a diet low in plant sterols that contained campestanol at about 2 mg/day. The heterozygote and three control subjects were fed a diet supplemented with a spread that contained campestanol at 540 mg/day and sitostanol (24-ethyl-5alpha-cholestan-3beta-ol) at 1.9 g/day as fatty acid esters. Plasma campestanol concentrations determined by capillary gas-liquid chromatography were 0.72 +/- 0.03 mg/dl in the homozygote, 0.09 +/- 0.04 mg/dl in the heterozygote, and 0.05 +/- 0.03 mg/dl for the control mean. After simultaneous pulse labeling with [3alpha-(3)H]campestanol intravenously and [23-(14)C]campestanol orally, the maximum percent absorption measured by the plasma dual-isotope ratio method as a single time point was 80% in the homozygote, 14.3% in the heterozygote, and 5.5 +/- 4.3% as the mean for three control subjects. Turnover (pool size) values estimated by mathematical analysis of the specific activity versus time [3alpha-(3)H]campestanol decay curves were as follows: 261 mg in the homozygote, 27.3 mg in the heterozygote, and 12.8 +/- 7.6 mg in the three control subjects (homogygote vs. controls, P < 0.001). The calculated production rate (mg/24 h) equivalent to actual absorption in the presence of dietary sterols and stanols was 0.67 mg/day or 31% of intake in the homozygote, 2.1 mg/day or 0.3% of intake in the heterozygote, and 0.7 +/- 0.3 mg/day or 0.1% of intake in the three control subjects. However, the excretion constant from pool A (K(A)) was prolonged markedly in the homozygote, but was 100 times more rapid in the heterozygote and three control subjects.Thus, campestanol, like other noncholesterol sterols, is hyperabsorbed and retained in sitosterolemic homozygotes. However, campestanol absorption was only slightly increased in the sitosterolemic heterozygote and removal was as rapid as in control subjects. PMID- 11060359 TI - Subcellular localization and physiological role of alpha-methylacyl-CoA racemase. AB - alpha-Methylacyl-CoA racemase plays an important role in the beta-oxidation of branched-chain fatty acids and fatty acid derivatives because it catalyzes the conversion of several (2R)-methyl-branched-chain fatty acyl-CoAs to their (S) stereoisomers. Only stereoisomers with the 2-methyl group in the (S) configuration can be degraded via beta-oxidation. Patients with a deficiency of alpha-methylacyl-CoA racemase accumulate in their plasma pristanic acid and the bile acid intermediates di- and trihydroxycholestanoic acid, which are all substrates of the peroxisomal beta-oxidation system. Subcellular fractionation experiments, however, revealed that both in humans and rats alpha-methylacyl-CoA racemase is bimodally distributed to both the peroxisome and the mitochondrion. Our findings show that the peroxisomal and mitochondrial enzymes are produced from the same gene and that, as a consequence, the bimodal distribution pattern must be the result of differential targeting of the same gene product. In addition, we investigated the physiological role of the enzyme in the mitochondrion. Both in vitro studies with purified heterologously expressed protein and in vivo studies in fibroblasts of patients with an alpha-methylacyl CoA racemase deficiency revealed that the mitochondrial enzyme plays a crucial role in the mitochondrial beta-oxidation of the breakdown products of pristanic acid byconverting (2R,6)-dimethylheptanoyl-CoA to its (S)-stereoisomer. PMID- 11060360 TI - Coronal acetabular fractures: the anterior approach in computed tomography navigated minimally invasive percutaneous fixation. AB - PURPOSE: To demonstrate the technical feasibility of the anterior approach to the coronal roof component of carefully selected acetabular fractures in computed tomography (CT)-navigated closed reduction and percutaneous fixation (CRPF). METHODS: Four patients with nondisplaced or slightly displaced coronal fractures of the acetabular roof were treated with percutaneous screw fixation. Screws were implanted over guidepins placed under CT navigation. Mean clinical and radiological follow-up was 16 months. RESULTS: All screws could be placed as intended. There were no peri- or postoperative complications. Radiological follow up showed primary osseous union. Clinical results were excellent according to a median Merle-d'Aubigne score of 18. CONCLUSION: Nondisplaced or slightly displaced coronally oriented fractures of the acetabular roof can be treated by minimally invasive percutaneous CT-navigated fixation through an anterior approach that does not endanger the sciatic nerve. Early clinical results are encouraging. Close cooperation between trauma surgeons and radiologists and careful selection of cases is mandatory. PMID- 11060361 TI - Retrospective comparison of the patency of Wallstents and Palmaz long-medium stents used for TIPS. Transjugular intrahepatic portosystemic shunts. AB - PURPOSE: To compare patency rates of transjugular intrahepatic portosystemic shunts (TIPS) after placement of long-medium Palmaz stents or Wallstents. METHODS: We performed a retrospective review of TIPS performed at our institution between December 1997 and December 1998. During this time period we placed long medium Palmaz stents for TIPS procedures in 17 patients and Wallstents in 20 patients as the initial stent. Patency was determined on follow-up by ultrasound, angiography, or pathologic examination in the event of transplant. RESULTS: Primary patency in the Palmaz stent group was 70.6% (12/17 patients) (follow-up 1 399 days, mean 127 days). Both primary assisted and secondary patency in the Palmaz group was 100% (17/17 patients) (follow up 1-399 days, mean 154 days). Primary patency in the Wallstent group was 50% (10/20 patients) (follow up 1-370 days, mean 65 days). Primary assisted patency in the Wallstent group was 80% (16/20 patients) (follow up 1-601 days, mean 141 days). Secondary patency in the Wallstent group was 100% (20/20 patients) (follow up 2-601 days, mean 142 days). Kaplan-Meier analysis of the two groups of patients yielded a primary patency of 266 days (standard error 45 days) for TIPS with the Palmaz stent and 139 days (standard error 45 days) for the Wallstent (p =.04). The 3, 6, and 12-month primary patency rates were .84, .63, and .42 respectively for the Palmaz stents and .36, .36, and .18 respectively for the Wallstent. There was no significant difference in primary assisted or secondary patency between the two stent groups. The mean tract curvature in the patients with Palmaz stents was 23.5 degrees (SD 18.2 degrees, range 0-69.0 degrees ) compared with 57 degrees (SD 34.5 degrees, range 7.0-144.0 degrees ) in patients with Wallstents (p =.01). CONCLUSIONS: Our nonprospective, nonrandomized study suggests that TIPS created with the long medium Palmaz stent have a higher primary patency than those created with the Wallstent in tracts that are relatively straight. PMID- 11060362 TI - Doppler flow wire evaluation of renal blood flow reserve in hypertensive patients with normal renal arteries. AB - PURPOSE: To study the vasomotor responses of the renal microcirculation in patients with essential hypertension. METHODS: We studied the reactivity of the renal microcirculation to papaverine, with intraarterial Doppler and quantitative arteriography, in 34 renal arteries of 19 hypertensive patients without significant renal artery stenosis. Isosorbide dinitrate was given to maximally dilate proximal renal arteries. APV (average peak blood flow velocity) was used as an index of renal blood flow. RESULTS: Kidneys could be divided into two distinct subgroups based on their response to papaverine. An increase in APV of up to 55% occurred in 21 kidneys, an increase > 55% in 13 kidneys. Within each group the values were normally distributed. Both baseline APV and the effect of papaverine on mean velocity differed significantly between groups. CONCLUSION: There seems to be a subgroup of patients with essential hypertension that has an impaired reactivity to papaverine, consistent with a functional impairment of the renal microcirculation. Further studies are required to determine whether this abnormality contributes to or results from elevated blood pressure. PMID- 11060363 TI - Better visualization of transbronchial biopsy using CT fluoroscopy. AB - PURPOSE: Computed tomography fluoroscopy (CTF) provides the capability for continuous CT imaging and has been increasingly used in interventional procedures. Our objective was to assess the usefulness of CTF in the monitoring of transbronchial biopsy procedures. METHODS: We evaluated nine patients in whom yield of "conventional" transbronchial biopsies had failed. CTF was performed on a Somatom Plus 4 Power scanner (CARE Vision CT, Siemens, Forchheim, Germany) using 120 kV, 50 mA at a frame rate of eight images per second on a matrix of 256 x 256. Image reconstruction was based on a partial scan with an acquisition time of 0.5 sec. The maximal time without interruption was 79 sec; after stopping for a few seconds a new period of 79 sec was available. The number of biopsies, procedure times, applied dose, and histologic results were documented. RESULTS: With CTF-guided transbronchial biopsy, the yield of the biopsies was improved. In seven patients biopsy yielded bronchial cancer; in one patient histopathologic examination showed tuberculosis. Only in one patient did CTF-guided transbronchial biopsy fail. The mean number of biopsies was four in each patient. Mean fluoroscopy time was 165 +/- 92 sec (range 111-272 sec) and mean procedure time was 800 +/- 302 sec (range 480-1081 sec). The applied dose ranged between 500 and 1224 mSv; the mean applied dose was 743 +/- 414 mSv. There were no fatal complications. CONCLUSION: Computed tomography fluoroscopy appears to facilitate visualization of transbronchial biopsy procedures, with the drawback of increased radiation exposure. To compare the "conventional" method versus CTF a randomized prospective study is necessary. PMID- 11060364 TI - Bronchial artery embolization for hemoptysis due to benign diseases: immediate and long-term results. AB - PURPOSE: To clarify the immediate effect and long-term results of bronchial artery embolization (BAE) for hemoptysis due to benign diseases and the factors influencing the outcomes. METHODS: One hundred and one patients (aged 34-89 years) received bronchial artery embolization with polyvinyl alcohol particles and gelatin sponge for massive or continuing moderate hemoptysis caused by benign pulmonary diseases and resistant to medical treatment. RESULTS: After BAE, bleeding stopped in 94 patients (94%). The immediate effect was unfavorable in cases where feeder vessels were overlooked or the embolization of the intercostal arteries was insufficient. Long-term cumulative hemoptysis nonrecurrence rates after the initial embolization were 77.7% for 1 year and 62.5% for 5 years. In bronchitis (n = 9) and active tuberculosis (n = 4) groups, an excellent (100%) 5 year cumulative nonrecurrence rate was obtained. The rate was lower in groups with pneumonia/abscess/pyothorax (n = 8) or with pulmonary aspergillosis (n = 9) (53.3%, 1-year cumulative nonrecurrence). There were higher incidences of early recurrence among patients with massive hemorrhage or more marked vascularity and systemic artery-pulmonary artery shunt in angiography: however, these trends were not statistically significant CONCLUSIONS: BAE can yield long-term benefit in patients with hemoptysis due to benign diseases. Technical problems in the procedure had an impact on the short-term effect. The degree of hemorrhage or the severity of angiographical findings were not significant factors affecting the outcome. The most significant factor affecting long-term results was whether the inflammation caused by the underlying disease was medically well controlled. PMID- 11060365 TI - Graft distortion after endovascular repair of abdominal aortic aneurysm: association with sac morphology and mid-term complications. AB - PURPOSE: To investigate the incidence, significance, and mechanism of stent-graft distortion after endovascular repair (EVR) of abdominal aortic aneurysm. METHODS: EVR of abdominal aortic aneurysm was performed in 51 cases (49 modular, bifurcated; 2 tube). Thirty-two patients were followed for 6 or more months and had equivalent baseline and follow-up images which could be used to determine changes in graft configuration. Sac dimensions were measured using computed tomographic (CT) images and graft-related complications were recorded. RESULTS: Amongst 32 patients evaluated on follow-up, there was graft distortion in 24. Distorted grafts were significantly (p = 0.002) associated with sac diameter reduction (mean 5 mm) and sac length reduction (mean 8.1 mm). All graft-related complications occurred in the limbs of eight distorted grafts, with a mean reduction of sac length in this group of 7.8 mm on reformatted CT images. CONCLUSION: There was a highly significant association between graft distortion and limb complications, and reduced sac dimensions. PMID- 11060366 TI - Long-term follow-Up of percutaneous balloon angioplasty in adult aortic coarctation. AB - PURPOSE: To assess long-term outcomes following percutaneous transluminal angioplasty (PTA) of congenital aortic coarctation in adults. METHODS: Seventeen patients underwent PTA for symptomatic adult coarctation of the aorta. Sixteen patients, with a mean age of 28 years (range 15-60 years), were reviewed at a mean interval after angioplasty of 7.3 years (range 1.5-11 years). Assessment included magnetic resonance imaging (MRI), Doppler echocardiography, and clinical examination. Current clinical measurements were compared with pre- and immediate post-angioplasty measurements. RESULTS: At follow-up 16 patients were alive and well. The patient not included in follow-up had undergone surgical repair and excision of the coarctation segment following PTA. Mean brachial systolic blood pressure for the group decreased from 174 mmHg before angioplasty to 130 mmHg at follow-up (p = 0.0001). The mean gradient had fallen significantly from 50.9 to 17.8 at follow-up (p = 0.001). The average number of antihypertensive drugs required per patient decreased from 0.56 to 0.31 (p = 0.234). No significant residual stenoses or restenoses were seen at MRI. Small but clinically insignificant residual pressure gradients were recorded in all patients using Doppler echocardiography. Complications included one transient ischemic attack at 5 days, one external iliac dissection requiring stent insertion, and a further patient who developed a false aneurysm close to the coarctation site at 12 months which subsequently required surgical excision. CONCLUSION: PTA of adult coarctation is safe and effective in the long term. Although primary stenting has recently been advocated in the treatment of this condition, our results suggest that PTA remains the treatment of choice. PMID- 11060367 TI - MR-Guided PTA in experimental bilateral rabbit renal artery stenosis and MR angiography follow-up versus histomorphometry. AB - PURPOSE: To assess in vivo 1) MR-guided percutaneous transluminal renal angioplasty (PTRA) in experimental bilateral rabbit renal artery stenosis (RAS); 2) postprocedural follow-up by gadolinium-enhanced MR angiography versus histomorphometry. METHODS: Fifteen male NZW rabbits of mean weight 4.0 kg (range 3.5-4.2 kg) underwent bilateral RAS induction by combined overdilation deendothelialization with a gadolinium-filled balloon, passively MR-guided by the artifact of a 0.014-inch guidewire. After 4 weeks the rabbits were randomized into two groups: group A (n = 8) underwent right-sided PTRA for treatment of RAS, group B (n = 7) underwent left-sided PTRA. After another 4 weeks the rabbits were killed to assess by histomorphometry recurrent stenosis and contralateral induction injury stenosis lesions. Each step was preceded by gadolinium-enhanced three-dimensional MR angiography, and the cortex-to-aorta (C/A) signal intensity ratio was calculated. RESULTS: RAS induction was successful in all cases. Fourteen arteries developed restenosis and 13 only initial stenosis. MR-guided PTRAs were feasible in 22 arteries (73%). For a successful catheterization of the ostium (20 arteries, 66% success rate), 10-25 steps were required. Five to eight steps were required for balloon localization and inflation for each PTRA. The restenosis effect was reflected by a 16% (12%-27%) decrease in C/A values on MR angiograms (p < 0.05). CONCLUSION: MR guidance and MR angiography represent a feasible, less invasive alternative for performing and assessing experimental PTRA in RAS. PMID- 11060368 TI - Assessment of a polyester-covered nitinol stent in the canine aorta and iliac arteries. AB - PURPOSE: To evaluate the patency and healing characteristics of a woven polyester fabric-covered stent in the canine model. METHODS: Twenty-four self-expanding covered stents were placed in the infrarenal aorta and bilateral common iliac arteries of eight dogs and evaluated at 1 (n = 2), 3 (n = 2), and 6 (n = 4) months. Stent assessment was done using angiography prior to euthanasia, and light and scanning electron microscopy. RESULTS: Angiographically, just prior to euthanasia, 8 of 8 aortic and 14 of 16 iliac endovascular covered stents were patent. Histologically, the stented regions showed complete endothelialization 6 months after graft implantation. A neointima had formed inside the stented vessel regions resulting in complete encasement of the fabric-covered stent by 3 months after graft implantation. Medial compression with smooth muscle cell atrophy was present in all stented regions. Explanted stent wires, examined by scanning electron microscopy, showed pitting but no cracks or breakage. CONCLUSION: The covered stent demonstrated predictable healing and is effective in preventing stenosis in vessels 10.0 mm or greater in diameter but does not completely preclude stenosis in vessels 6.0 mm or less in diameter. PMID- 11060369 TI - Percutaneous transcatheter aortic disc valve prosthesis implantation: a feasibility study. AB - PURPOSE: Over the past 30 years there have been experimental efforts at catheter based management of aortic valve regurgitation with the idea of extending treatment to nonsurgical candidates. A new catheter-based aortic valve design is described. METHODS: The new catheter-delivered valve consists of a stent-based valve cage with locking mechanism and a prosthetic flexible tilting valve disc. The valve cage is delivered first followed by deployment and locking of the disc. In acute experiments, valve implantation was done in four dogs. RESULTS: Valve implantation was successful in all four animals. The implanted valve functioned well for the duration of the experiments (up to 3 hr). CONCLUSION: The study showed the implantation feasibility and short-term function of the tested catheter-based aortic disc valve. Further experimental studies are warranted. PMID- 11060370 TI - Percutaneous ablation of an internal iliac aneurysm using tissue adhesive. AB - We report the percutaneous injection of tissue adhesive (Tisseal, Immuno, Vienna, Austria) to ablate a 12-cm internal iliac aneurysm. The complex history of this lesion included previous surgery for a ruptured aortic aneurysm, attempted repair of the internal iliac aneurysm, and several embolization procedures. These factors precluded further open repair or transcatheter techniques and dictated the choice of a more direct approach. PMID- 11060371 TI - Successful transcatheter embolization of penetrating aortic ulcer using interlocking detachable coils. AB - A 54-year-old man with persistent chest pain was hospitalized for hypertension and DeBakey type IIIb aortic dissection. The false lumen of the dissection was almost completely thrombosed; however, a penetrating atherosclerotic ulcer (PAU) was observed 5 weeks later. At that time, we successfully embolized the PAU with a microcatheter and interlocking detachable coils (IDCs). The patient is well with no episodes of relapse in 20 months of follow-up. This case suggests the utility of the microcatheter and IDC system as an alternative to surgery. PMID- 11060372 TI - Anastomoses of the ovarian and uterine arteries: a potential pitfall and cause of failure of uterine embolization. AB - Four women with symptomatic uterine fibroids were treated by uterine artery embolization (UAE). In all cases both uterine arteries were embolized via a single femoral puncture with polyvinyl alcohol using a selective catheter technique. In three cases, the ovarian artery was not visible on the initial angiogram before embolization, but appeared after the second uterine artery had been treated. In one case of clinical failure following UAE, a repeat angiogram demonstrated filling of the fibroids from the ovarian artery. Anastomoses between uterine and ovarian arteries may cause problems for radiologists performing UAE and are a potential cause of treatment failure. PMID- 11060373 TI - Thrombin injection for treatment of brachial artery pseudoaneurysm at the site of a hemodialysis fistula: report of two patients. AB - We report two patients with arteriovenous hemodialysis fistulas that were complicated by brachial artery pseudoaneurysms. Each pseudoaneurysm was percutaneously thrombosed with an injection of thrombin, using techniques to prevent escape of thrombin into the native brachial artery. In one patient, an angioplasty balloon was inflated across the neck of the aneurysm during thrombin injection. In the second patient, thrombin was injected during ultrasound-guided compression of the neck of the pseudoaneurysm. Complete thrombosis of each pseudoaneurysm was achieved within 30 sec. No ischemic or embolic events occurred. This technique may be useful in treating pseudoaneurysms of smaller peripheral arteries. PMID- 11060374 TI - Portal hypertension secondary to spontaneous arterio-portal venous fistulas: transcatheter arterial embolization with n-butyl cyanoacrylate and microcoils. AB - We report a 73-year-old man with recurrent variceal bleeding due to portal hypertension caused by multiple intrahepatic arterio-portal venous fistulas, which were successfully occluded by embolization with n-butyl cyanoacrylate and micro-coils. PMID- 11060375 TI - Retrieval of a Greenfield IVC filter displaced to the right brachiocephalic vein. AB - During insertion of a central venous sheath an inferior vena cava stainless steel Greenfield filter was dislodged to the right brachiocephalic vein without a free end. Successful retrieval was achieved by using a combination of a guidewire and a snare. Percutaneous retrieval of this vena cava filter is feasible with minimal risk using this method. PMID- 11060376 TI - Adjuvant therapy in the treatment of complications following surgery for hepatic echinococcal cysts. AB - Thirty-two patients had surgery for hepatic echinococcal cysts (HEC). Serious complications were observed in 16 patients (50%): cyst recurrence (n = 4), infected residual cyst cavity (n = 7), infected residual cyst cavity with biliary and duodenal fistulae (n = 2), recurrent biliary obstruction following open surgery for a ruptured HEC into the biliary tree (n = 2), delayed rupture of an HEC into the biliary tree following laparoscopic surgery with secondary biliary obstruction (n = 1). These major complications were successfully managed by percutaneous methods in 8 of 16 patients while antihelmintic therapy was sufficient in two patients with a small recurrent cyst and ERCP was used in one patient to relieve biliary obstruction. Surgery was required in two patients only. With a success rate of 87.5%, the nonsurgical approach is the preferred method for treating a post-surgical complication. PMID- 11060377 TI - Re: jejunal artery angioplasty and coronary stent placement for acute mesenteric ischemia. PMID- 11060379 TI - Announcements PMID- 11060378 TI - Re: the safe use of cetrimide and hypertonic saline for percutaneous ablation of hepatic echinococcal cysts complicated by intrabiliary rupture. PMID- 11060380 TI - [Isotretinoin and pregnancy]. PMID- 11060381 TI - [Neonatal lupus erythematosus (NLE). Towards a better knowledge of the physiopathology]. PMID- 11060382 TI - [Isotretinoin in childbearing women: compliance with strengthen warnings]. AB - INTRODUCTION: The recommendations for prescription and dispensing of Roaccutane (isotretinoid) were strengthened in 1997 in order to reduce the number of pregnancies exposed to Roaccutane. The aim of this study was to evaluate the incidence of exposed pregnancies since this time and the compliance with the new recommendations. MATERIAL AND METHODS: All pregnancies exposed to Roaccutane reported to French Regional Drug Monitoring Centers, to Laboratoire Roche or to the Information Center for Teratogenic Agents since the publication of these recommendations for prescription were studied (March 1997-December 1998). In addition, compliance with the new recommendations was evaluated by sampling 169 drug prescriptions dispensed at 105 pharmacies in France. RESULTS: Thirty-seven pregnancies were exposed to Roaccutane during the risk period because of failure of contraceptive methods (28 p. 100), contraception incorrectly followed (52 p. 100) or not prescribed (20 p. 100). The incidence of pregnancies exposed to Roaccutane during the risk period evaluated at 0.6/1,000 women of child-bearing age [0.4-0.8] is very close to the incidence reported in the earlier study which prompted the new recommendations. Thirty-three percent of the 169 prescriptions studied did not carry all the legal warnings. Roaccutane was correctly prescribed for only 18 p. 100 of women, i.e. with a contraceptive method as recommended by the French Drugs Monitoring Agency and with full, correct information. Although the most important recommendations had been followed, 12 p. 100 of women were treated with Roaccutane without contraception and 16 p. 100 received a contraceptive method not recommended by the French Drugs Monitoring Agency (in particular Diane). DISCUSSION: The provision of information must be further improved by enforcing the modalities of prescription and dispensing of Roaccutane. However, it appears that there is no way of completely avoiding patient-related failure. PMID- 11060383 TI - [Cutaneous neonatal lupus erythematosus: discordant expression in identical twins]. AB - BACKGROUND: Neonatal lupus erythematosus is a rare syndrome characterized essentially by cutaneous lesions and/or congenital heart block occurring in infants at birth, or shortly after. It is related to transplacental crossing of maternal auto antibodies (usually anti Ro/SS-A, La/SS-B or rarely anti-U(1) RNP) from the mother to the infant. Mothers of affected children have signs of systemic lupus erythematosus or other collagenosis or are asymptomatic. CASE REPORT: We report a case of neonatal lupus erythematosus in one identical twin, revealed at the age of 3 months by erythematous and annular cutaneous lesions of the face and limbs. These lesions were preceded at birth by an asymmetrical livedo pattern of the lower limbs. Her twin sister was unaffected but both infants had a high rate of anti-Ro/SSA antibodies. The diagnosis of neonatal lupus erythematosus permitted to reveal a biological lupus syndrome in their asymptomatic mother. Cutaneous lesions cleared almost completely within 1 year whereas antiRo/SSA antibodies disappeared. CONCLUSION: Cases of neonatal lupus erythematosus in twins are rare and mostly described in heterozygotic twins. Clinical discordance is usual and could partly be explained by genetic factors. In monozygotic twins, like in our case, chromosome X inactivation could be an explanation of the differences observed. PMID- 11060384 TI - [Medium-term renal involvement in adult patients with Henoch-Schonlein purpura]. AB - INTRODUCTION: The aim of this study was to estimate the frequency of medium-term renal involvement, in a series of adult patients with Henoch-Schonlein purpura treated in a dermatology department. PATIENTS AND METHODS: Seventeen patients with Henoch-Schonlein purpura followed from 1991 to 1997 in the department of dermatology were included in the study. Renal tests included: research of microscopic hematuria, proteinuria, plasma creatinine levels and calculated creatinine clearance which were measured during initial hospitalization and at the date of the study in May 1998. RESULTS: 10 men and 7 women (mean-age 51 years) were followed up for 39 months (6 to 79 months). Initial renal involvement was observed in 11 patients (65 p. 100). A microscopic hematuria was also observed in 9 patients (53 p. 100), a proteinuria in 6 (35 p. 100), an association of hematuria and proteinuria in 4 (23.5 p. 100). No patient had renal failure. In May 1998, only 2 patients (11.7 p. 100) had renal involvement, that consisted of proteinuria: 3.3 and 3. 9 g/d respectively, with no renal failure. DISCUSSION: The 65 p. 100 frequency of renal involvement discovered during the acute phase of HSP in our series was similar to other series mentioned in the literature, which mainly consisted of hematuria and/or proteinuria. The frequency of long-term renal involvement depends on the origin of patient recruitment. Nephrological studies have reported high levels of renal involvement, that probably overestimated the true frequency. In the present study, as well as in two other studies from a non-nephrological recruitment, the frequency of long term renal involvement was estimated between 11 p. 100 and 16 p. 100 and was primarily persistent proteinuria. CONCLUSION: The frequency of renal involvement in the adult patient with HSP during the acute phase of the disease and after a medium-term follow-up was approximately 50 p. 100 and between 11 p. 100 to 16 p. 100 respectively. PMID- 11060385 TI - [Muckle-Wells syndrome: 4 cases in three generations]. AB - BACKGROUND: Muckle-Wells syndrome is a hereditary condition with variable penetrance. The main manifestations are urticarial rash, malaise in the evening, joint pain, perception deafness and renal amylosis. CASE REPORT: We describe a family with 4 affected members in 3 successive generations. Clinical expression was variable. DISCUSSION: Despite the absence of renal amylosis in our patients, this family presented the syndrome described by Muckle and Wells in 1962. As for other cases reported in the literature, the clinical course was favorable with low-dose corticosteroid therapy. PMID- 11060386 TI - [Cheyletiella dermatitis: an uncommon cause of vesiculobullous eruption]. AB - BACKGROUND: Species of Cheyletiella mites are parasites hosted by dogs, cats and rabbits. In humans, they cause a dermatosis not well known by dermatologists. We report a case of an unusual, purely vesiculobullous eruption due to Cheyletiella blakei acquired from an infected cat. CASE REPORT: A 76-year-old woman presented a very pruritic eruption of vesicles and bullous lesions located on the trunk and external areas of the arms. Biopsy showed dermoepidermal cleavage and polynuclear infiltrate with prominent eosinophils, consistent with the diagnosis of bullous pemphigoid. We suspected a Cheyletiella dermatitis due to the aspect and distribution of the elementary lesions and the fact that prior to the eruption the patient had acquired a cat that sometimes slept in her bed. The diagnosis was confirmed by a veterinary examination and isolation of Cheyletiella blakei from the cat's skin. The cat was treated successfully with ivermectin, while the household was disinfected with permethrin. A treatment with benzyl benzoate (Ascabiol) resolved all the patient's symptoms. DISCUSSION: This case is particularly interesting due to the purely vesiculobullous pattern of the rash and by the difficulty and considerable delay of diagnosis. Patients who have recently acquired a cat or dog before developing a pruritic dermatosis may indeed have cheyletiellosis. PMID- 11060387 TI - [Solar urticaria: one case treated by intravenous immunoglobulin]. AB - BACKGROUND: Solar urticaria is a rare photodermatosis which often begins from the third to the fifth decade. Usual treatment consists of photoprotection measures and antihistamines although disease control is sometimes unsatisfactory with both. We report herein a very severe case of solar urticaria we treated with intravenous immunoglobulins. CASE-REPORT: A 55-year-old woman suffered for 3 years from very severe solar urticaria which resisted treatment. Phototests revealed two action spectra: the first in UVA near 380 nm with a minimal urticarian dose of 0.025 J/cm(2), the second near 500 nm in visible light. RESULTS: As last resort treatment, we gave our patient intravenous immunoglobulins. After the third course of treatment, the improvement was impressive as the patient could tolerate visible light and 15 minutes of intense solar exposure. The minimal urticarian dose was raised from 0.025 J/cm(2) to 27 J/cm(2) in UVA. One year after treatment, the solar urticaria has disappeared. CONCLUSIONS: We report herein the first case of solar urticaria treated with success with immunoglobulin. Intravenous immunoglobulin treatment is well for its effectiveness in many autoimmune diseases such as autoimmune thrombocytopenic purpura, and also, as recently proven, in some cases of severe idiopathic chronic urticaria. PMID- 11060388 TI - [Rhabdomyosarcoma at site of pacemaker implantation]. AB - BACKGROUND: Malignant proliferation at the site of implantation of a pacemaker generator is uncommon. We report the case of a patient who developed rhabdomyosarcoma. CASE REPORT: A 85-year-old man presented with a voluminous and rapidly evolving tumor localised beneath the right clavicle. This inflammatory and necrotic lesion developed on the area where a titanium pacemaker had been implanted five years earlier. Rhabdomyosarcoma was diagnosis on the basis of immunohistochemistry findings. In spite of a wide surgical excision of this primitive tumour, visceral dissemination appeared, rapidly leading to the patient's death. DISCUSSION: This observation rises the question of the role of the pacemaker implantation in tumor development. The excellent in vivo tolerance and the widespread utilization of titanium as biomaterial is an argument against its carcinogenic action. Inversely, a metal-related chronic inflammatory reaction could favor the neoplastic process in predisposed subjects as has been observed with prosthetic materials used in orthopaedics. PMID- 11060389 TI - [Minocycline hypersensibility syndrome]. PMID- 11060391 TI - [Cutaneous leiomyomas and leiomyosarcomas and Epstein-Barr virus]. PMID- 11060392 TI - [Pseudomonas aeruginosa toe web infections: successful treatment by ointment polymyxine B - oxytetracycline (Primyxine)]. PMID- 11060393 TI - [Pyoderma gangrenosum in the French West Indies (Martinique): 7 cases]. PMID- 11060394 TI - [An umbilical lesion. Isolated umbilical polyp]. PMID- 11060395 TI - [Ulceration of the plantar vault. Ecthyma gangrenosum]. PMID- 11060396 TI - [Dermatologic care for minimal acute burns]. PMID- 11060397 TI - [Dynamic phototherapy for skin cancer and precancer lesion]. PMID- 11060398 TI - [Biological diagnosis of porphyria cutanea tarda]. PMID- 11060399 TI - [How useful is the study of the T-cell repertoire in cutaneous T-cell lymphomas?]. PMID- 11060400 TI - [Bullae induced by acute neuroleptic intoxication]. PMID- 11060401 TI - [Mondor's disease]. PMID- 11060402 TI - [Acarbose (Glucor) and severe polymorphous erythema]. PMID- 11060404 TI - [Pruritus probios: ad pad] [In Process Citation] PMID- 11060403 TI - [The Library of the Karolinska Institutet in Stockholm]. PMID- 11060405 TI - [Immunomodulator structure-activity relationships: contribution of molecular modeling]. AB - Molecular modeling used to compare 64 immunostimulant compounds with pyrrolie quinolein or purine nuclei has pointed out that a common spatial structure is found in most of the active compounds. An additional study of immunostimulants (levamisole, muramyldipeptide) or immunosuppressive molecules (rapamycin) was performed. A common pharmacophore was found on every studied compound. It was composed of three neighboring electroattractive atoms and a further fourth atom. The favorable conformation of rapamycin for immunosuppressive action, which is not the more stable conformation, could explain the loss of its activity, or those of related macrolides, when some minor chemical modifications are tested. These findings validate the proposed concept and provide a view of the mechanism of action of most of the immunomodulator compounds for preparing novel compounds PMID- 11060406 TI - [Study of lipid peroxidation inhibition in human blood by several endogenous and exogenous compounds]. AB - Numerous studies suggest that lipid peroxidation is involved in the atherosclerosis and hemolytic disease processes. Nicanartine improves in vitro resistance of LDL (low density lipoproteins) to oxidation in the conjugated dienes model. Polarographic assays show that hemin-bound drugs inhibit erythrocyte membrane peroxidation. A method to measure the antioxidant capacity of plasma is proposed and tested in sickle cell disease. These in vitro results suggest drugs and drugs combination which could be efficient to inhibit lipid peroxidation in vivo. PMID- 11060407 TI - [Rigorous algorithms for calculating the exact concentrations and activity levels of all the different species during acid-base titrations in water]. AB - The principles of two algorithms allowing the calculations of the concentration and activity levels of the different species during acid-base titrations in water are described. They simulate titrations at constant and variable ionic strengths respectively. They are designed so acid and base strengths, their concentrations and the titrant volume added can be chosen freely. The calculations are based on rigorous equations with a general scope. They are sufficiently compact to be processed on pocket calculators. The algorithms can easily simulate pH-metric, spectrophotometric, conductometric and calorimetric titrations, and hence allow determining concentrations and some physico-chemical constants related to the occurring chemical systems. PMID- 11060408 TI - [Synthesis and biological evaluation of indole derivatives acting as anti inflammatory or antitumoral drugs]. AB - Two axes of research have been explored, one about promising non-acidic non steroidal anti-inflammatory derivatives, with indolin-2-one as structural core and another one about aromatase inhibitors, characterized by azolylmethyl or alpha-azolylbenzyl chain on indole nucleus. Knoevenagel reaction led to indolin-2 ones substituted by either 2,6-di-tert-butylphenol chain or 1, 4-dihydropyridine chain, revealing antioxydant or anti-inflammatory activities. Aromatase is a logical target in the treatment of hormono-dependent breast cancer in postmenopausal women. Among non steroidal inhibitors of this enzyme, diverse compounds with anilino or azaheterocyclic moiety are currently used or undergoing clinical trials. Our pharmacomodulation in azolylmethylindole or alpha azolylbenzylindole series led to compounds with high level aromatase inhibitory activity. Work to determine their selectivity by measuring their inhibitory effect on P450 17alpha enzyme was also carried out. A first molecular modeling approach with Discover software was performed to evaluate interactions between our molecules and the catalytic site of P450cam. PMID- 11060409 TI - [Recognizing interventional biology as a specialty on its own]. AB - Changing therapeutic strategies have led to the development of new techniques where elements of the human body, including cells, tissues or even organs, are removed from the body in order to modify their potential before reintroducing them into the body. The ex vivo manipulations are performed by biologists working in close coordination with clinicians who determine the indications and perform the procurement and reintroduction steps. We propose the term interventional biology for this type of activity and suggest that, beyond the various domains considered including reproduction biology, and cell and gene therapy, it should be recognized as a specific form of practice of medical biology. Recognized as a specialty on its own, this type of activity could be better organized helping improve training, rules of good practice, performance assessment, and establishment of individual responsibilities. This would provide the means for guaranteeing efficacy and safety for a rapidly growing field of activity. PMID- 11060410 TI - [The subgenus Paraphlebotomus (Phlebotomus--Phlebotominae-- Psychodidae- Diptera): a review. Morphological and molecular studies]. AB - A systematic review of the subgenus Paraphlebotomus (a group of phlebotomine sandflies which contains several vectors of leishmaniasis) is proposed. In the morphological approach, we study the available types and a lot of sandflies coming from different geographic areas. The taxonomic status of each species is revised. We consider P. marismortui as a synonym of P. alexandri. We suppose that P. mofidii is a valid species. P. similis is clearly differentiated from P. sergenti. A new species is described from North Africa: P. riouxi. We discuss the validity of P. saevus. Two molecular phylogenies based on ribosomal DNA sequences are proposed. The first one used the D2 domain of the 28 S sub-unit. It emphasises a very close relation between the subgenera Paraphlebotomus and Phlebotomus, a likely paraphyly of the genus Phlebotomus and the position of the genera Sergentomyia and Lutzomyia as two sister groups. The second one used the ITS 2. It emphasises the informative capacity of this gene for the resolution of the phylogenetic relations within the subgenera Paraphlebotomus. PMID- 11060411 TI - [Flavonoid and dianthranoid levels of the St.-John's-wort flowering tops]. AB - Dried flowering tops of Hypericum perforatum L. (16 batches) and H. maculatum (3 batches) were studied according to the harvest period. The mean levels of the flavonoid and dianthranoid compounds were respectively: total flavonoids 3.20 and 3.92%, hyperoside 0.87 and 2. 12%, rutin 0.54 and 0.02%, isoquercitrin 0.48 and 0.75%; total dianthranoids 0.14 and 0.13%, hypericin 0.04 and 0.02%, pseudohypericin 0.08 and 0.11%. The flavonoid and dianthranoid levels were higher in young flowering tops of H. perforatum. 3 commercial batches were also examined for a comparative study. Pharmacopoeial specifications are proposed for a revision of the monograph "St.-John's-wort". PMID- 11060412 TI - [Extrarenal salt excretion in oceanic birds]. AB - Oceanic birds feed mainly on marine invertebrates which are isosmotic with sea water. The kidneys of birds have a low concentrating ability and are unable to produce urine with a sodium concentration more than 300mmol/l. How do these birds avoid dehydration? The excess salt is eliminated by the secretory work of a paired gland in the head known for centuries to anatomists as nasal glands or supraorbitary glands, and now that their function is elucidated called salt excreting glands or simply salt glands. They connect through a duct with the nasal cavity. Oceanic birds expel by the nares a clear and colourless watery liquid that contains primarily Na(+) and Cl(-), at high and constant concentration, with a small amount of K(+) and HCO(3)(-). These salt glands do not function continually as the kidney does, becoming active only in response to an osmotic load. The salt gland of oceanic birds is one of the most efficient systems of ionic transport in the living world. PMID- 11060413 TI - [Vanadium and diabetes: pancreatic and peripheral insulinomimetic properties] [In Process Citation] AB - A very large body of research has been devoted to diabetes mellitus with the goal of better understanding this complex disease for better patient treatment. Pathophysiological research in the rat has focused on Pharmacological research suggests that vanadium could be a possible therapeutic agent due to its activity on insulin secretion and its peripheral insulinomimetic properties. Results suggest that the antidiabetic properties of vanadium result from Vanadium derivatives thus comprise a novel class of compounds with promising therapeutic potential and favorable pharmacokinetic properties (oral administration). PMID- 11060414 TI - [ [In Process Citation] PMID- 11060415 TI - [A century of progress. From sutures to arterial reconstruction]. PMID- 11060416 TI - [Intravascular ultrasound: limitation and prospects]. AB - The aims of intravascular ultrasound (IVUS) are visualization of the arterial wall, characterization of the arterial plaque, measurement of vessel diameter and endoluminal procedure assessment. The main sources of errors are: limitation of resolution, distortion of images due to non uniform angular velocity of mechanical type transducers, overestimation or underestimation of cross sectional area secondary to eccentric position or non coaxial orientation of the catheter. Awareness of these problems may prevent misinterpretations. Combined balloon imaging catheters, forward-looking sector scanners and ultrasound imaging guide wires may be achieve by miniaturization of transducers. Three dimensional imaging, blood flow quantification, local elasticity of tissues may be achieve by new softwares. PMID- 11060417 TI - [Blood and arterial wall rheology and cardiovascular risk factors]. AB - The vascular endothelium and circulating blood cells are exposed to a hemodynamic environment related to the pulsatile nature of the pressure and blood flow which influence the morphology, the physical properties and the metabolism of the cells. Among these mechanical forces, shear stresses, related to the speed gradients and to the blood viscosity, control vascular vasomotor tone and thrombogenecity, stimulating the production of the endothelial factors of relaxation or contraction, of the coagulation factors or activating erythrocyte and platelet aggregation or disaggregation phenomena. Low shear stress is considered to be a critical factor in the causation of thickening of the arterial wall and in the formation of atheromatous plaques. These plaques develop predominantly in specific parts of the arterial tree where asymmetries in the velocity profiles occur. A close relationship has been found in a large general population between plasma viscosity and thickening of the carotid bifurcation. The blood cells share the same environment as the endothelial cells. Thus, the shear stresses to which the erythrocytes are submitted is a major determinant of the blood viscosity. Shear is also an important physiological parameter regulating platelet aggregation in flowing suspensions. However, in vivo, the response of platelets depends on the balance between activation of the platelets by shear and the same rheological forces acting on the endothelial cells which produce mediators which inhibit this activation in order to maintain blood fluidity. PMID- 11060418 TI - [Rationalization of risk factors for venous thromboembolism in medical inpatients. A prospective study]. AB - BACKGROUND: In terms of preventive management of venous thromboembolism in medical inpatients, very large differences may be observed. Rationalization of behaviour requires the evaluation of simple and logical parameters, which takes into account both patient safety and economic considerations. AIM: The aim of this study was to evaluate a preventive scheme including the rationalization of the indications and the use of low molecular weight heparin. METHODS: EXPERIMENTAL DESIGN: Epidemiologic investigation. SETTING AND PATIENTS: Patients hospitalized in five medical departments in the Hospital Center of Nantes, France. INTERVENTION: The risk of venous thromboembolism was rated as high, intermediate and low. Patients with high or intermediate risk were eligible for prevention therapy (table I). MEASURES: The main criterion was the occurrence during hospital stay of deep or superficial venous thrombosis of the lower limbs, pulmonary embolism, or unexplained sudden death. The screening was based on clinical features double-checked by venous doppler ultrasonography of the lower limbs and/or ventilation-perfusion lung scanning. RESULTS: 24,497 patients were eligible (table II), 15% were considered at risk and treated with Nadroparin, 6% had the same risk profile but were not treated and 14. 7% had low risk and no prevention. No bleeding event was reported. The incidence of venous thromboembolism was 0.75%, 1.7% and 0.14% respectively (p <0.01) (table III). This efficacy does not appear to depend on body weight or the existence of multiple risk factors observed (table IV and V). CONCLUSIONS: This analysis of risk factors separates two populations with rates of incidence dramatically and significantly different. The prevention of venous thromboembolism by fixed dose of low molecular weight heparin remains justified since it reduces the risk of venous thromboembolism by a factor of 2.5. PMID- 11060419 TI - [Upper extremity deep venous thrombosis. 40 hospitalized patients]. AB - Deep venous thrombosis is 50 times less frequent in upper than in lower limbs. Data remain poor in the literature. Forty consecutive patients (24 males, 16 females, mean age: 54.5 years) were retrospectively analysed from 161 subjects who underwent venous explorations of the upper extremity for a 3.5 year period in the same center. Diagnosis of thrombosis was made by duplex ultrasonography (n =37) or phlebography (n =3). Main clinical manifestations were edema (n =36) and pain (n =29). Location of thrombosis was humeral (n =1), axillary (n =2), or sub clavian (n =37, 2 bilateral). The majority of thrombosis (n =29) were secondary to cancer and venous catheter (n =19, 15 implanted ports), to central catheter alone (n =3) or cancer alone (n =7). The 11 others were associated with thoracic outlet syndrome (n =6) or apparent primary thrombosis (n =5). Thrombophilia was identified in 6 out of these 11. During follow up [mean of 9 months (0,5-36)], two patients developed pulmonary embolism, 14 a post-thrombotic syndrome and 16 patients died. Initial therapy included heparin (n =36) or fibrinolysis (n =4). Upper extremity deep venous thrombosis are mostly associated with cancers and venous catheters. Thrombophilia is frequent in the other cases. Heparin followed by oral anticoagulation is the optimal therapy whose duration depends upon underlying condition. Fibrinolysis has not been useful for preventing post thrombotic syndrome in our study. PMID- 11060420 TI - [Placebo controlled efficacy of class 1 elastic stockings in chronic venous insufficiency of the lower limbs]. AB - OBJECTIVE: To compare, in a 4-week clinical trial, the efficacy of class 1 Elastic Compression Stockings (pressure at the ankle 10 to 15 mmHg) to that of Placebo Stockings (pressure at the ankle 3 to 6 mmHg) in women consulting in general practice for mild, symptomatic, chronic venous insufficiency of the lower limbs. PATIENTS AND METHODS: This was a 4-week multicentre, randomised, double blind, placebo-controlled clinical trial conducted on two parallel groups of patients presenting with mild chronic venous insufficiency grade C(1-3S) E(p) A(S1-5) according to the CEAP classification. Treatment efficacy was assessed by the following criteria: global impairment as assessed at each visit on a visual analogue scale (primary efficacy parameter), Quality of Life measured by the CIVIQ questionnaire, symptoms index (sum of individual scores for pain, limb heaviness, paresthesias, cramps and evening limb oedema), limb volume measured by volumetry, and the evolution of global impairment during the course of the trial as assessed by repeated auto-evaluations on visual analogue scales. Compliance was assessed by the number of days patients wore the stockings for at least ten hours per day. Treatment tolerability was assessed by the record of the possible adverse events. RESULTS: Three hundred and forty one patients were included in total. Statistically significant differences were observed on global score of quality of life in favour of class 1 Elastic Compression Stockings group. In the subgroup of patients with evening limb oedema at inclusion, the decrease of limb volume noted with class 1 Elastic Compression Stockings was all the more marked that evening limb oedema was more pronounced at baseline. A good compliance was observed in 76% to 80% of patients according to the treatment group. Tolerability of class 1 Elastic Compression Stockings was comparable to that of the Placebo Stockings. CONCLUSIONS: After four weeks of treatment, compression with class 1 Elastic Compression Stockings allows a statistically significant improvement of Quality of Life and a decreasing of limb oedema in patients presenting with mild chronic venous insufficiency of the lower limbs. Moreover, the tolerability of class 1 Elastic Compression Stockings was confirmed by the high level of patient compliance. PMID- 11060421 TI - [Abdominal aortic aneurysm repair treated with endoprosthesis: technical and economic evaluation by ANAES(National Agency of Health Accreditation and Evaluation)]. AB - OBJECTIVE: To evaluate the safety, efficacy and cost of stent-grafts for the endovascular repair of abdominal aortic aneurysms (AAA). MATERIALS AND METHODS: The ANAES report was based on a critical analysis of the published international literature and on consultation of technology assessment agencies. The report was reviewed by a group of experts and validated by the ANAES Scientific Council (Evaluation Section). RESULTS: The literature analysis indicated that the technique was feasible. However, it also revealed a shortage of methodologically sound clinical trials. The long-term outcome of the stent-grafts and the accompanying complications are not yet known. CONCLUSION: Further clinical studies are needed for a truly objective assessment. The ANAES working group has issued a series of recommendations designed to help physicians define the optimal conditions for the effective and safe use of these stent-grafts. PMID- 11060422 TI - [Consensus conference. Erysipelas and necrotizing fasciitis. Short text]. PMID- 11060423 TI - [Spontaneous internal carotid artery dissection]. AB - Spontaneous carotid artery dissection is one of the main causes of stroke in young adults. We report two cases occurring after non-traumatic hyperextension of the head. Most of these events result from minor head trauma. Some family history cases would suggest the possibility of an underlying artery wall disorder. The favorable natural history of carotid dissection emphasizes the need for a noninvasive approach for diagnosis. Delayed diagnosis may lead to poor prognosis. Ultrasonography and magnetic resonance angiography are adequate for diagnosis. PMID- 11060424 TI - [Kaposiform angiodermatitis (Bluefarb-Stewart syndrome) caused by a superficial arteriovenous malformation of the trochanter area: a case report]. AB - We reported a case of kaposiform angiodermatitis (Bluefarb-Stewart syndrome) complicating a superficial arterio-venous malformation of the skin and the sub cutaneous tissue of the right trochanter area of a 28 year-old-man. This lesion resulted from large arterio-venous shunts occurring over a one-year period inside the vascular malformation, which remained stable for 27 years. This case report underlines that any vascular malformation may take an uncertain turn and needs an attentive follow-up with adequate treatment. PMID- 11060425 TI - [A "forgotten" first in surgery: resection of an aortic arch aneurysm, Olivier Monod, Paris, 1947]. PMID- 11060426 TI - [Perfusion imaging of a dynamic scanner: contribution to functional assessment of renal artery stenosis]. PMID- 11060427 TI - [What is the role of scintigraphy in the diagnosis of suspected renovascular renal insufficiency?]. PMID- 11060428 TI - [Long-term influence of associated arthrodesis on adjacent segments in the treatment of lumbar stenosis: a series of 127 cases with 9-year follow-up]. AB - PURPOSE OF THE STUDY: Little is known about the impact of posterolateral arthrodesis on adjacent levels. In order to examine this question, we analyzed the radiological evolution of the lumbar spine in patients treated for lumbar stenosis, comparing cases where posterolateral arthrodesis was used with the other cases. Our aim was to determine whether the long-term radiographical modifications were affected by the arthrodesis. MATERIAL AND METHODS: Among our series of patients presenting with lumbar stenosis between 1984 and 1992, we retained two groups: patients in group 1 (n=46) who underwent single-level decompressions at L4-L5 or L4-L5 and L5-S1 level; and patients in group II (n=81) who underwent decompressions on the same levels associated with posterolateral arthrodesis extending from L4 to the sacrum with or without instrumentation. We compared the course of the two levels above the decompression (L2-L3 and L3-L4) between the two groups. We compared three radiological parameters: disc height, intervertebral slipping, and intersegmental mobility. We also examined the correlations between radiological modifications and functional outcome. Mean follow-up for these 127 patients was 9 years. RESULTS: The two groups were comparable for age, gender, follow-up, and presurgical functional score, disc height and intervertebral slipping at equivalent levels. At last follow-up, disc narrowing was observed at L2-L3 and L3-L4; it was significantly greater in the group with complementary arthrodesis. At L3-L4, intervertebral slipping also worsened more in the arthrodesis patients. Use of osteosynthesis significantly increased the risk of developing such radiological lesions. These lesions were associated, solely in the arthrodesis group, with poorer functional outcome. CONCLUSION: Our findings allow the conclusion that, despite the effect of physiological aging, the observed long-term degenerative lesions in patients undergoing treatment of lumbar stenosis are related to the associated arthrodesis which increases their frequency and severity, deteriorating the functional outcome. PMID- 11060429 TI - [Bankart procedure: clinical and radiological long-term outcome]. AB - PURPOSE OF THE STUDY: There are few reports onlong-term outcome after Bankart procedure. The purpose of this study was to determine the rate of recurrent dislocation, the clinical results and the incidence of glenohumeral osteoarthritis after a minimum 10-year follow-up. MATERIAL AND METHODS: Ninety seven Bankart procedures were performed in 97 patients between 1972 and 1986 for treatment of anterior shoulder instability with recurrent dislocations. We retrospectively reviewed 74 patients and obtained 64 complete radioclinical evaluations for an average follow-up of 16 years. Clinical evaluation was based on the G. Walch and the Duplay group score but for easier comparisons, we also calculated the Rowe et al. score. Radiographical evaluation was established on the Samilson and Prieto classification but real glenohumeral osteoarthritis with joint narrowing was noted independently as grade four. We also studied the contralateral shoulder. RESULTS: At last follow-up, 7 shoulders (9.5%) had recurrent dislocation, but two of them occurred subsequent to severe trauma over 18 months. Most patients (95 %) were satisfied or very satisfied. Six patients (8.1%) had persistent apprehension but in some it was not due to anterior apprehension. According to the Duplay score (or the Rowe score), 25 shoulders (44.6%) had an excellent result (35/61.4 %) 16 (28.6%) a good result (7/12.3%), 11 (19.7%) a fair result (11.19.3) and 4 (5.4 %) a poor result (4/7%). Operated shoulders were pain free for 75% and painful for forced movements only for 25%. External rotation at 90 degrees of abduction was reduced by 8.7 +/- 15.7 degrees. There was no limitation of internal rotation. Patients returned to preoperative sports activities at the same level for 70.9 % and at a lower level for 12.7%. According to the Samilson classification, 7 (13%) of the shoulders had grade 2 and 2 (3.7%) had grade 3 glenohumeral osteoarthritis. We found 4 cases (7.4%) of real glenohumeral osteoarthritis (grade four) and 2 of these patients had contralateral osteoarthritis of a non unstable shoulder. There was no perioperative complication. DISCUSSION: In our hands the Bankart procedure is appeared as a safe procedure with a low rate of glenohumeral osteoarthritis and a high rate of patient satisfaction. PMID- 11060430 TI - [Arthroscopic rotator cuff repair: a multicentric retrospective study of 87 cases with anatomical assessment]. AB - PURPOSE OF THE STUDY: The aim of this study was to evaluate the anatomical and technical limits of endoscopic rotator cuff repair. Anatomical results were also compared with functional assessment of the shoulder. MATERIAL AND METHODS: A multicentric serie of 87 patients presenting a full thickness rotator cuff tear repaired endoscopically was retrospectively reviewed at 25.4 months (mean) post surgery. Limits of the surgical technique were studied in correlation with functional results and anatomic radiographic evaluation (arthroscans in 93 p. 100). RESULTS: Anatomical repair (normal thickness and no contrast in the subacromial space on arthroscan) was achieved in 83 p. 100 of the rotator cuffs with limited damage to the frontal part of the supra spinatus tendon. This percentage fell to 57.8 p. 100 in case of posterior extension of the tear to the supra spinatus tendon and further dropped to 40.8 p. 100 in case of retraction to the apex of the humeral head. Functional outcome evaluated with the Constant score was strongly related to the radiographic cuff condition (p <0.05). For distal and anterior tears of the supra spinatus tendon, the Constant score at revision was 89.8 points in cases with anatomic repair at revision. This score fell to 75 when the rotator cuff tear was evidenced radiographically (p <0.0001). For tears involving the entire supra spinatus tendon repaired by arthroscopy, the functional difference at revision was 8 points on the Constant scale. Objective and subjective analysis of the surgical procedure identified significant peroperative elements predictive of clinical and anatomical outcome (difficult reduction, p <0.05; subjective degree of solidity, p <0.0001; anatomical aspect of the repaired cuff, p <0.05). DISCUSSION: A comparison of our findings with data on equivalent lesions reported in the literature suggests that endoscopic surgery for rotator cuff tears offers both functional and anatomic results equivalent to those achieved with conventional open surgery. This assumes that the surgical procedure is carried out by surgeons experienced in shoulder arthroscopy who can precisely gauge the posterior/anterior extension of the tears and the limits of the surgical technique. PMID- 11060431 TI - [Congenital pseudarthrosis of the clavicle: 25 childhood cases]. AB - PURPOSE OF THE STUDY: Congenital pseudarthrosis of the clavicle is rare, only 200 cases having been reported. Based on 25 personal cases and an overview of the literature, we try to explain the etiology of this condition and the different kinds of treatment. MATERIAL AND METHODS: A retrospective analysis was performed on twenty-five children (16 females, 9 males, mean age at the end of the follow up - 11.5 yrs) from three different centers. We assessed the outcome of surgical and nonsurgical procedures, based on pain, functional ability, cosmetic results, and x-ray examination. RESULTS: A family background was noted in three children. The lesion always involved the right side. Twenty patients presented a bump over the middle third of the clavicle, thirteen a foreshortened shoulder girdle, three complained of discomfort. In two cases, palpation of the clavicular area was painful. No neurovascular compressive syndrome was reported. None of the patients complained of a decrease in the range of motion or in the strength of the upper limb. X-rays showed a middle third defect. In five cases we found abnormal first ribs. Seventeen patients underwent surgery, at a mean age of 6 years and 4 months. The procedure always included excision of the pseudarthrosis at both ends and internal fixation with a wire or a plate. In only eight cases a bone graft was used. Healing was achieved in fourteen patients. Three patients needed a second surgical procedure. In these 3 cases we had not used bone grafting. All patients had a normal range of shoulder motion, except a twelve year old girl who complained of discomfort of the right upper limb. The cosmetic result was good in eleven cases, one surgical wound was noted as hypertrophic, and one developed a keloid. An asymmetry of the trunk was still noted in seven cases. The x-rays showed symmetric clavicles in ten cases. Eight patients were treated conservatively. All of them had a normal range of motion of the shoulder, six had a good cosmetic result and two cases a poor one. DISCUSSION: According to Alldred, the anomaly results from the failed coalescence of the two primary ossification centers of the clavicle. The overview of the literature and our findings (in one case) confirm that the cartilage which covers both ends of the bone is made of growth cartilage. However, the true mechanism of the nonunion is still unknown. The three familial cases of our work suggest a possible genetic transmission of the disease. The diagnosis is based on the following criteria: right side lesion, found in infancy, without previous fracture, increasing size with growth, without major functional consequences, without neurofibromatosis or cleidocranial dysostosis symptom. X-rays or histologic examination will confirm the diagnosis showing the usual findings described above. Complications of the pseudarthrosis of the clavicle are rare and late. Conservative management appears to give good results as seen with our eight patients. However surgical treatment ensures symmetrical shoulder girdles and good function with few complications. Therefore, we recommend performing an excision of the cartilaginous caps, followed by an iliac bone graft and an internal fixation with wire. Surgical management will be preferred in symptomatic patients, in the case of major or increasing deformity, or on parental request. PMID- 11060432 TI - [Functional gait adaptations in patients with painful hip]. AB - PURPOSE OF THE STUDY: This prospective study was conducted to analyze the mechanisms of gait compensation in patients with painful hip and to search for correlations with preoperative clinical and radiographic findings. MATERIAL AND METHODS: Optoelectronic and multicomponent force-plate datas were used to calculate joint motion, moments and intersegmental forces for 26 patients with unilateral hip pain and 20 normal age and sex-matched patients. Height was similar in the two groups but mean weight in the study group (83 kg) was greater than in the controls (68 kg). The preoperative Harris score was 53 in the study group and 16 patients had a permanent flexion contracture of the knee (mean 15 degrees, range 5-30 degrees). Radiographically, there were 22 cases of osteo arthritis hip disease and 4 cases of necrosis. RESULTS: Gait analysis showed a significant 0.66 +/- 0.06 m (12 p. 100) reduction in step length. Patients who had severe hip pain walked with a decreased dynamic range of motion (18 +/- 5 degrees, p<0.0001) with a curve reversal as they extended the hip. They also reduced dynamic range of motion of the knee and ankle. Patients who presented a reversal in their dynamic hip range of motion had a greater passive flexion contracture and a greater loss in range of motion during gait than those with a smooth regular pattern (p<0.0001). Patients with hip pain walked with significantly decreased external extension, adduction, and internal and external rotation moments (p<0.0001). They also unloaded the ipsilateral knee and ankle. The decreased hip extension moment was significantly correlated with an increased level of pain (p<0.0001). There was no correlation with radiological findings. DISCUSSION: Reversal of dynamic hip range of motion was interpreted as a mechanism to increase effective hip extension during stance phase through increased anterior pelvic tilt and lumbar lordosis. CONCLUSION: Patients with painful hip walked with a manner that was asymmetric. These gait modifications were related to hip limitation in passive motion and pain. Patients with flexion contracture adopted a compensatory gait mechanism. This study confirms relation between hip pain and forces across the hip joint. PMID- 11060433 TI - [Noncemented total hip arthroplasty: influence of extramedullary parameters on initial implant stability and on bone-implant interface stresses]. AB - PURPOSE OF THE STUDY: After total hip replacement, the initial stability of the cementless femoral stem is a prerequisite for ensuring bone ingrowth and therefore long term fixation of the stem. For custom made implants, long term success of the replacement has been associated with reconstruction of the offset, antero/retro version of the neck orientation and its varus/valgus orientation angle. The goals of this study were to analyze the effects of the extra-medullary parameters on the stability of a noncemented stem after a total hip replacement, and to evaluate the change of stress transfer. MATERIAL AND METHODS: The geometry of a femur was reconstructed from CT-scanner data to obtain a three-dimensional model with distribution of bone density. The intra-medullary shape of the stem was based on the CT-scanner. Seven extra-medullary stem designs were compared: 1) Anatomical case based on the reconstruction of the femoral head position from the CT data; 2) Retroverted case of - 15 degrees with respect to the anatomical reconstruction; 3) Anteverted case with an excessive anteversion angle of + 15 degrees with respect to the anatomical case; 4) Medial case: shortened femoral neck length (- 10 mm) inducing a medial shift of the femoral head offset; 5) Lateral case: elongated femoral neck length (+ 10 mm) inducing lateral shift of the femoral head offset 6) Varus case with CCD angle 127 degrees; 7) Valgus case with CCD angle 143 degrees. The plasma sprayed stem surface was modeled with a frictional contact between bone and implant (friction coefficient: 0.6). The loading condition corresponding to the single limb stance phase during the gait cycle was used for all cases. Applied loads included major muscular forces (gluteus maximus, gluteus medius, psoas). RESULTS: Micromotions (debonding and slipping) of the stems relative to the femur and interfacial stresses (pressure and friction) were different according to the extra-medullary parameters. However, the locations of peak stresses and micromotions were not modified. The highest micromotions and stresses corresponded to the lateral situation and to the anteverted case (micro-slipping and pressure were increased up to 35 p.100). High peak pressure was observed for all designs, ranging from anatomical case (34 MPa) to anteverted case (44 MPa). The peak stresses and micromotions were minimal for the anatomical case. The maximal micro-debonding was not significantly modified by the extra-medullary design of the femoral stem. DISCUSSION: The extra medullary stem design has been shown to affect the primary stability of implant and the stress transfer after THR. Most interfacial regions present small micro slipping which normally allows the occurrence of bone ingrowth. The anatomical design presents the lowest micromotions and the lowest interfacial stresses. The worst cases correspond to the anteverted and lateralized cases. Probably, the anteverted situation involves higher torsion torque, which in turn may induce high torsion shear micromotions and higher stress at the interface. Moreover, the lever arm of the weight bearing force on the femoral head is augmented for the augmented neck length situation. This increases the bending moment, and therefore may increase the stresses as well as the stem shear micromotions. In summary, the present results could be taken as biomechanical arguments for the requirement of anatomical reconstruction of not only the intra-medullary shape but also the extra-medullary parameters (reconstruction of the normal hip biomechanics). PMID- 11060434 TI - [Risks and results after simultaneous intramedullary nailing in bilateral femoral fractures: a retrospective study of 40 cases]. AB - PURPOSE OF THE STUDY: A retrospective series of 40 patients who underwent simultaneous intramedullary nailings for bilateral femoral shaft fractures was analyzed. The aim of our study was to verify that simultaneous nailing without reaming does not increased risk of fat embolism and to assess clinical and radiological outcome. MATERIAL AND METHOD: This series included 27 men and 13 women, mean age 27.8 years, who underwent first intention intramedullary nailing between 1986 and February 1999. Thirty-two patients had multiple fractures. Mean ISS was 23 (range 9 to 59). Among the 80 femoral shaft fractures, 15 were open fractures, 3 were associated with sciatic paralysis, and 4 were complicated by an interruption of the femoral vessels. The AO classification was: type A=44; type B=25; type C=11. Mean delay to simultaneous centromedullary nailing was 3. 8 days: surgery was performed on the day of arrival for 25 patients. General anesthesia was used in all cases with respiratory assistance (FIO(2) =50 to 100 p. 100). Mean nail diameter was 11.6 (range 10-14). Gurd criteria and PaO(2) were followed to assess pulmonary function. Clinical and radiological outcome was assessed using the modified Thorensen criteria. RESULTS: Preoperatively, PaO(2) was< 87 mmHg in 8 patients. Four of these patients showed a discrete drop off and three improved well above the normal level. Only one patient experienced an important decrease but did not develop respiratory distress. Among the 32 patients with a normal level preoperatively, PaO(2) remained in the normal range in 18, fell to a limit level but below 87 mmHg in 4, and showed a substantial drop off of 46 to 172 mmHg in 10. Two of these 10 patients developed respiratory distress due to fat embolism which was fatal in one case. One other patient died in the immediate postoperative period of an undetermined cause. All of the other patients recovered normal gas levels within a few hours or days. There were four cases of phlebitis, including one with pulmonary embolism, one case of respiratory distress by pulmonary superinfection, and one case of septicemia. Both femoral fracture sites became infected in one patient. Malunion occurred in two cases. Two vascular repairs of the femoropopliteal axis were unsuccessful, leading to above knee amputations. Thirty-four patients have been examined after a minimal 12 months follow-up (mean 30 months). Outcome was excellent for 48 femurs, good for 10 and fair for 10. DISCUSSION: This continuous series of simultaneous bilateral femoral shaft intramedullary nailings appears to be the only such report to date. The clinical and radiological outcomes were comparable with those achieved in one-side femoral fractures. The risk of fat embolism is inevitable after long bone fractures. Many factors favoring the risk are recognized, the most important being delay to fixation. Reaming creates excessive pressure in the medullary canal and could thus contribute to the risk. The presence of an associated chest trauma is not a formal contraindication if effective hematosis is preserved as evidenced by the blood gases. CONCLUSION: Simultaneous nailing of bilateral femoral shaft fractures can be performed if blood gases remain acceptable and minimal reaming is used. PMID- 11060435 TI - [Osteochondral lesions of the talar dome: surgical treatment in a series of 30 cases]. AB - PURPOSE OF THE STUDY: We reviewed 30 cases of osteochondral lesions of the astragalar vault treated surgically. MATERIAL AND METHODS: Among the 30 patients, 17 participated in sports activities and 24 had a history of trauma. Mean delay to surgery was 10 months. Treatment included osteochondritis curettage and Pridie perforations. Direct access was used in 11 cases, malleolar osteotomy in 13 and arthroscopy in 6. Cancellous bone grafts were used in 6 cases. RESULTS: Mean follow-up was 3 years 7 months (minimum 2 years). All patients had an arthroscan at last follow-up. Evaluation of post-operative outcome was based on clinical assessment and arthroscan findings. Surgical treatment provided very good results in 75 p. 100 of cases with pain relief and improved walking distance. DISCUSSION: Our cases pointed out the important contribution of the FOG (Fracture Osteonecrosis Geode) classification to pathogenic and prognostic analysis. The Berndt and Harty classifications were not found to be useful. CONCLUSION: In case of localized necrosis, we propose arthroscopic perforation curettage. In case of bone loss, a direct cancellous graft may be used. PMID- 11060436 TI - [Aneurysmal bone cyst of the patella. A case report and literature review]. AB - We report a case of an aneurysmal cyst localized in the patella of a 37-year-old man. The lesion was secondary to a chondroblastoma at six years follow-up after initial curettage and bone graft. It were no recurrence. Treatment of aneurysmal cysts depends on the degree of articular involvement. We made a detailed study of 11 cases of this rare localization of aneurysmal cysts reported in the literature. PMID- 11060437 TI - [False popliteal aneurysm after tibial osteotomy: a case report]. AB - A 55-year-old man developed a pseudoaneurysm of the popliteal artery after tibial valgization osteotomy performed for degenerative genu varum. A tourniquet was used for the procedure. A wedge osteotomy was performed two centimeters under the joint line; the correction angle was ten degrees. Immediately after the end of the procedure, the distal pulses disappeared for ten minutes. Doppler exploration of the arterial network did not demonstrate any anomaly. Ten days postoperatively, the patient complained of sudden onset pain in the knee and tension in the popliteal fossa. Arteriography demonstrated a pseudo-aneurysm of the popliteal artery. The lesion caused an interruption of arterial flow and was successfully treated by emergency resection and suture. PMID- 11060438 TI - Report of the third international workshop on human chromosome 10 mapping and sequencing 1999. PMID- 11060439 TI - Mini review: form and function in the human interphase chromosome. AB - A key feature of interphase chromosomes is their compaction into discrete "territories" in the nucleus. In this review, we focus on the compartmentalization of the genome conferred by this organization and evaluate our current understanding of the role of large-scale chromatin folding in the regulation of gene expression. We examine evidence for the hypothesis that transcription occurs at the external surfaces of chromosomes and follow its evolution to include transcription at the surfaces of chromatin-rich domains within chromosomes. We also present prevailing views regarding the details of large-scale chromatin folding and the functional relationship between chromatin and the enigmatic nuclear matrix. PMID- 11060440 TI - Cloning of leptin cDNA and assignment to the long arm of chromosome 5 in the marsupial Sminthopsis crassicaudata. AB - We have isolated and sequenced full-length cDNA clones for leptin in the dasyurid marsupial Sminthopsis crassicaudata (fat-tailed dunnart). Southern and in situ hybridisation data indicated a single leptin gene in the S. crassicauda- ta genome, localised to arbitrary chromosome bands 5q24--> q31 on the long arm of chromosome 5, the short-arm terminus of which bears the only nucleolar organising region. The nucleotide sequence of the cDNAs revealed that the primary translation product of S. crassicaudata leptin is composed of 167 amino acid residues, with a potential signal peptide of 21 residues. The mature protein of 146 amino acids is 82% similar to both the mouse and human proteins and is predicted to have a molecular weight of 16.26 kDa. Northern blot analysis revealed that the corresponding mRNA is approximately 3.9 kb in size and is expressed only in white adipose tissue of this marsupial species. Evolutionary analyses indicate that S. crassicaudata leptin cDNA has evolved at a significantly faster rate than cDNAs from eutherian mammals. PMID- 11060441 TI - Characterization of three hairy cell leukemia- derived cell lines (ESKOL, JOK-1, and hair-M) by multiplex-FISH, comparative genomic hybridization, FISH, PRINS, and dideoxyPRINS. AB - Hairy cell leukemia (HCL) is a chronic B-lymphocyte leukemia. Due to the proliferative inertia of the malignant cells, HCL-derived cell lines are an important tool for studies on this disease. We have elaborated the karyotypes of three HCL-derived cell lines, ESKOL, JOK-1, and Hair-M, by combining a range of molecular cytogenetic techniques, including multiplex (multicolor) fluorescence in situ hybridization (M-FISH), comparative genomic hybridization (CGH), conventional FISH, primed in situ labeling (PRINS), and dideoxyPRINS. We found ESKOL to be monoclonal with a single chromosome aberration, der(7)t(3;7)(q26.3;q31). JOK-1 also appeared to be monoclonal, having the karyotype 48,XY,der(4) t(1;4)(1pter-->p32::4qter-->pter), der(6)(6qter-->p22::q12 -> qter), +der(7)t(7;11)(7pter-->q21::11p15-->pter),der(8)t(5;8;12) (5pter- >p14::12p11.2-->p12::8q12-->q21::8?cen-->-->24 .?2), der(14)t(8;14)(8qter- >q24.?2::14q32.3-->pter),+20. These karyotypes differ from the original descriptions of ESKOL and JOK-1. The Hair-M cells analyzed by us were found to be peritetraploid with numerous chromosomal rearrangements. The cell line was also found to be multiclonal. On this basis, we do not regard the Hair-M cell line to be suitable for HCL studies. PMID- 11060442 TI - Differential increase of steroid sulfatase activity in XX and XY trophoblast cells from human term placenta with syncytia formation in vitro. AB - Steroid sulfatase (STS, EC 3.1.6.2) catalyzes the hydrolysis of the sulfate ester bonds of a variety of sulfated steroids, such as cholesterol, dehydroepiandrosterone, and estrone sulfate, a reaction influencing fertility and breast cancer in mammals. The activity of the enzyme is substantially elevated in placental syncytiotrophoblasts and is lower in other somatic cells. The polypeptide sequence of the enzyme is encoded by a gene located on the distal short arm of the human X chromosome. Prior studies have shown that the STS gene escapes X-chromosome inactivation. We studied the expression of the STS gene in primary cultures of cytotrophoblasts from human term placentae and compared it with the expression of autosomally encoded placental alkaline phosphatase (PALP) and X-linked glucose-6-phosphate dehydrogenase (G6PD). During 90 h in culture, the mononucleated cytotrophoblast cells did not proliferate, but differentiated into multinucleated, syncytiotrophoblast-like cells. STS activity in freshly isolated cytotrophoblasts was low (about 17%), compared to placental tis- sue, and about 1.7-fold higher in female than in male cells. During cultivation, STS activity increased 2- to 3-fold in female, but not in male, cells. PALP activity was very low in freshly isolated cytotrophoblasts (about 5% of placental tissue), and no significant difference between female and male cells was detectable. Within 90 h in culture, PALP activity increased in all preparations about 2- to 4 fold. G6PD activity in freshly isolated cytotrophoblasts showed great variation among preparations, and no significant difference between female and male cells was detectable. In both male and female cells the activity declined to about 50% of initial activity during cultivation. We conclude that human cytotrophoblasts in primary culture show a sex-specific regulation of STS activity, perhaps as a unique feature of the STS gene. The cytotrophoblast system may offer a new possibility to study the regulation of STS gene expression. PMID- 11060443 TI - Human secretin (SCT): gene structure, chromosome location, and distribution of mRNA. AB - Secretin is an endocrine hormone that stimulates the secretion of bicarbonate rich pancreatic fluids. Recently, it has been discussed that secretin deficiency may be implicated in autistic syndrome, suggesting that the hormone could have a neuroendocrine function in addition to its role in digestion. In the present study, the human secretin gene (SCT) was isolated from a bacterial artificial chromosome genomic library. SCT contains four exons, with the protein coding regions spanning 713 bp of genomic DNA. Human SCT is similar structurally to the secretin genes of other species. Amino acid conservation, however, is most pronounced within the exon encoding the biologically active mature peptide. Northern blot analysis shows that human SCT transcripts are located in the spleen, intestinal tract, and brain. Radiation hybrid mapping places the SCT locus on chromosome 11p15.5. PMID- 11060444 TI - Genetic and physical mapping of the porcine melatonin receptor 1B gene (MTNR1B) to chromosome 9. PMID- 11060445 TI - Assignment of the centromere protein H (Cenph) gene to mouse chromosome band 13D1 by in situ hybridization and interspecific backcross analyses. PMID- 11060446 TI - Assignment of karyopherin alpha 1 (KPNA1) to human chromosome band 3q21 by in situ hybridization. PMID- 11060447 TI - The human inward rectifier K(+) channel subunit kir5.1 (KCNJ16) maps to chromosome 17q25 and is expressed in kidney and pancreas. AB - A novel human Kir5.1 (inward rectifier K+ channel subunit, gene name KCNJ16) was identified through database searches. This human KCNJ16 was mapped to chromosome 17q25. The full-length cDNA was identified and its genomic structure was determined. Tissue distribution studies showed that human KCNJ16 is significantly expressed in human kidney, pancreas and thyroid gland. In situ hybridization revealed expression in convoluted tubule cells of kidney and in the acinar and ductal cells of pancreas. These suggest that human Kir5.1 may be involved in the regulation of fluid and pH balance, thus making it a potential therapeutic target for hypertension, renal failure, or pancreatic disease. PMID- 11060448 TI - Assignment of the MDM2 binding protein gene (MTBP) to human chromosome band 8q24 by in situ hybridization. PMID- 11060449 TI - Assignment of the canine paired-box 3 (PAX3) gene to chromosome 37q16-->q17 by in situ hybridization. PMID- 11060450 TI - Assignment of transcription factor NFAT5 to human chromosome 16q22.1, murine chromosome 8D and porcine chromosome 6p1.4 and comparison of the polyglutamine domains. AB - To date, transcription factors of the NFAT family (nuclear factors of activated T cells) have been described for mouse and man. Recently, we mapped the human NFAT5 gene to chromosome 16 by PCR using DNA from hybrid cell lines. Here we report the exact position of the human gene between D16S496 and WI5254 within the 16q22.1 subband, the localization of the murine gene at chromosome 8D, and the identification and mapping of the porcine counterpart to chromosome 6p1.4. PMID- 11060451 TI - Expression and comparative chromosomal mapping of MKP-5 genes DUSP10/Dusp10. AB - We have isolated a mouse cDNA for a novel MAPK phosphatase, designated as MKP-5. Two MKP-5 mRNA transcripts of 3.5 kb and 2.7 kb were detected. The 3.5-kb transcript was expressed in almost all the tissues examined, and was particularly abundant in cerebellum, skeletal muscle, and bone marrow. On the other hand, the 2.7-kb transcript was specifically and highly expressed in testis. The MKP-5 genes (DUSP10/Dusp10) were localized to chr 1q41, Chr 1H5 and chr 13q26 in human, mouse and rat, respectively. They were mapped in regions where conserved linkage homology has been identified among the three species. PMID- 11060452 TI - Molecular cloning, expression and chromosome location of the human pelota gene PELO. AB - The pelota gene of Drosophila melanogaster encodes a protein that was found to be included in cell cycle regulation. Mutations were found to result in spermatogenic arrest, female sterility and disturbances in the patterning of the eye. Here we describe the cloning of the human pelota cDNA (PELO) that encodes a 385-amino-acid protein. Southern blot and fluorescence in situ hybridization analyses revealed that PELO is present as a single copy gene in the human genome and is localized on chromosome 5q11.2. Northern blot analysis revealed the presence of a 1.6-kb transcript in all tissues studied and an additional 2.0-kb transcript in testis. PMID- 11060453 TI - Spontaneous frequencies of aneuploid and diploid sperm in 10 normal Chinese men: assessed by multicolor fluorescence in situ hybridization. AB - Many studies have been published establishing the background frequencies of disomic and diploid sperm in normal men by fluorescence in situ hybridization (FISH) analysis, with highly significant variance among the reports. Besides interdonor heterogeneity and differences in the experimental protocols used, the question of inherent differences in chromosome malsegregation and meiotic arrest among different geographic and ethnic groups of donors has been raised. In this study, multicolor FISH analysis was carried out on semen samples from 10 nonsmoking, nondrinking Chinese men from the People's Republic of China. The results were compared to FISH data on 10 nonsmoking, nondrinking Canadians under the same experimental conditions, in the same laboratory. A total of 200,497 sperm was scored in the Chinese donors and compared to 202,320 sperm from Canadian donors. Approximately 10,000 sperm per chromosome probe per donor were analyzed. The mean hybridization efficiency was 99.99%. The frequencies of X bearing and Y-bearing sperm were not significantly different from the expected 50% for each individual and for the combined data from all donors (49.73% vs. 49.46%, P = 0.3946). The mean disomy frequencies (range) were 0.07% (0.02%-0.12%) for chromosome 13, 0.18% (0.09%-0.19%) for chromosome 21, 0.05% (0. 01%-0.09%) for 24,XX, 0.02% (0.01%-0.06%) for 24,YY, and 0.29% (0. 13%-0.49%) for 24,XY. The mean diploidy frequency (range) was 0.38% (0.22%-0.73%) for 13-21 hybridizations and 0.32% (0.07%-0.70%) for XY hybridizations. Highly significant interdonor heterogeneity was found for diploidy (P = 0.0000) and for XY disomy (P = 0.0011), but no age effect was observed in any category of disomic or diploid sperm. The data reported here show no marked differences in disomy and diploidy frequencies between the mainland Chinese and Canadian groups, if donor heterogeneity is taken into account. PMID- 11060454 TI - Assignment of BCAT1, the gene encoding cytosolic branched chain aminotransferase, to sheep chromosome band 3q33 and to cattle and goat chromosome bands 5q33 by in situ hybridization. PMID- 11060455 TI - Detection of chromosomal imbalances in leiomyosarcoma by comparative genomic hybridization and interphase cytogenetics. AB - Leiomyosarcomas comprise a group of malignant soft-tissue tumors with smooth muscle differentiation. In this study, 14 cases of leiomyosarcoma were screened for changes in relative chromosome copy number by comparative genomic hybridization. A high number of imbalances (mean, 16.3; range, 6-26) was detected, with chromosomal gains occurring about twice as much as losses. The most frequent gains were found in 5p15, 8q24, 15q25-->q26, 17p, and Xp (43% to 50%), whereas the most frequent losses were found in 10q and 13q (50% and 78%, respectively). Twenty high-level amplifications affecting 15 different chromosomal subregions were detected in nine different tumors. In three leiomyosarcomas, sequences on chromosome arm 17p were found to be highly amplified, with a minimal overlapping region on subbands 17p12-->p11. We further discovered that the Smith-Magenis syndrome critical region on 17p11.2 is included in the 17p amplicons of two leiomyosarcoma cases. Using probes flanking this genetically unstable region, a mean of 14 and 22 signals per nucleus, respectively, was detected in both leiomyosarcomas by fluorescence in situ hybridization. In conclusion, this analysis identifies a number of characteristic chromosomal imbalances in leiomyosarcomas and provides evidence for the localization of potential oncogenes and tumor suppressor genes active in leiomyosarcoma genomes. PMID- 11060456 TI - DNA methylation and chromosome instability in lymphoblastoid cell lines. AB - In order to gain more insight into the relationships between DNA methylation and genome stability, chromosomal and molecular evolutions of four Epstein-Barr virus transformed human lymphoblastoid cell lines were followed in culture for more than 2 yr. The four cell lines underwent early, strong overall demethylation of the genome. The classical satellite-rich, heterochromatic,juxtacentromeric regions of chromosomes 1, 9, and 16 and the distal part of the long arm of the Y chromosome displayed specific behavior with time in culture. In two cell lines, they underwent a strong demethylation, involving successively chromosomes Y, 9, 16, and 1, whereas in the two other cell lines, they remained heavily methylated. For classical satellite 2-rich heterochromatic regions of chromosomes 1 and 16, a direct relationship could be established between their demethylation, their undercondensation at metaphase, and their involvement in non-clonal rearrangements. Unstable sites distributed along the whole chromosomes were found only when the heterochromatic regions of chromosomes 1 and 16 were unstable. The classical satellite 3-rich heterochromatic region of chromosomes 9 and Y, despite their strong demethylation, remained condensed and stable. Genome demethylation and chromosome instability could not be related to variations in mRNA amounts of the DNA methyltransferases DNMT1, DNMT3A, and DNMT3B and DNA demethylase. These data suggest that the influence of DNA demethylation on chromosome stability is modulated by a sequence-specific chromatin structure. PMID- 11060457 TI - Thirteen type I loci from HSA4q, HSA6p, HSA7q and HSA12q were comparatively FISH mapped in four river buffalo and sheep chromosomes. AB - Thirteen goat BAC clones containing coding sequences from HSA7, HSA12q, HSA4 and HSA6p were fluorescence in situ mapped to river buffalo (Bubalus bubalis, BBU) and sheep (Ovis aries, OAR) R-banded chromosomes. The following type I loci were mapped: BCP to BBU8q32 and OAR4q32, CLCN1 to BBU8q34 and OAR4q34, IGFBP3 to BBU8q24 and OAR4q27, KRT to BBU4q21 and OAR 3q21, IFNG to BBU4q23 and OAR3q23, IGF1 to BBU4q31 and OAR3q31, GNRHR to BBU7q32 and OAR6q32, MTP to BBU7q21 and OAR6q15, PDE6B to BBU7q36 and OAR6q36, BF to BBU2p22 and OAR20q22, EDN1 to BBU2p24 and OAR20q24, GSTA1 to BBU2p22 and OAR20q22, OLADRB (MHC) to BBU2p22 and OAR20q22. All mapped loci appeared to be located on homologous chromosomes and chromosome bands in both bovids. Comparison between gene orders in bovid (BBU and OAR) and human (HSA) chromosomes revealed complex rearrangements, especially between BBU7/OAR6 and HSA4, as well as between BBU2p/OAR20 and HSA6p. PMID- 11060458 TI - Isolation and characterization of a new FHL1 variant (FHL1C) from porcine skeletal muscle. AB - Four and a half LIM domain protein 1 (FHL1) was initially described as an abundant skeletal muscle protein with four LIM domains and a GATA like zinc finger. FHL1 was shown to be expressed in skeletal muscle as well as in a variety of other tissues. Recently, alternatively spliced FHL1 mRNAs were identified coding for C-terminal truncated proteins. The tissue distribution of these variants is more restricted and their functional properties seem to be different. We have isolated and characterized a new variant of FHL1 from porcine skeletal muscle (FHL1C). FHL1C is characterized by a newly identified start codon resulting in a 16 amino acids longer N- terminal region. We have isolated and characterized the porcine FHL1C gene spanning approximately 14 kb and harboring six exons. Using primer extension analysis, the transcription start site of FHL1C was mapped, indicating that FHL1C is regulated by an alternative promoter. The tissue distribution of FHL1C expression was studied by RT-PCR. The porcine FHL1C gene was assigned to the distal part of the long arm of the X chromosome by fluorescence in situ hybridization and screening of a somatic porcine/rodent cell hybrid panel. PMID- 11060459 TI - Genomic structure and chromosome mapping of human and mouse RAMP genes. AB - The cDNAs for human and murine Receptor Activity Modifying Proteins and for the associated murine Calcitonin Receptor Like Receptor were isolated. The human RAMP1 and RAMP3 genes possess two introns and human RAMP2 possesses three introns. Human RAMP1 was assigned to chromosome 2q36-->q37.1, RAMP2 to 17q12- >q21.1 and RAMP3 to 7p13-->p12. Mouse Ramp1 was assigned to chromosome 1 and Ramp2 and Ramp3 were assigned to chromosome 11. PMID- 11060460 TI - No deleterious mutations in the FOXJ1 (alias HFH-4) gene in patients with primary ciliary dyskinesia (PCD). AB - The transcription factor FOXJ1 (alias HFH-4 or FKHL13) of the winged helix/forkhead family is expressed in cells with cilia or flagella, and seems to be involved in the regulation of axonemal structural proteins. The knockout mouse Foxj1(-/-) shows abnormalities of organ situs, consistent with random determination of left-right asymmetry, and a complete absence of cilia. The human FOXJ1 gene which maps to chromosome 17q, is thus an excellent candidate gene for Kartagener Syndrome (KS), a subphenotype of Primary Ciliary Dyskinesia (PCD), characterized by bronchiectasis, chronic sinusitis and situs inversus. We have collected samples from 61 PCD families, in 31 of which there are at least two affected individuals. Two families with complete aciliogenesis, and six families, in which the affected members have microsatellite alleles concordant for a locus on distal chromosome 17q, were screened for mutations in the two exons and intron exon junctions of the FOXJ1 gene. No sequence abnormalities were observed in the DNAs of the affected individuals of the selected families. These results demonstrate that the FOXJ1 gene is not responsible for the PCD/KS phenotype in the families examined. PMID- 11060461 TI - Assignment of microrchidia (Morc) to mouse chromosome 16 by interspecific backcross linkage analysis and human chromosome 3q13 using somatic cell hybrids and in situ hybridization. PMID- 11060462 TI - Molecular cloning of Xp11 breakpoints in two unrelated mentally retarded females with X;autosome translocations. AB - Mental retardation is a very common and extremely heterogeneous disorder that affects about 3% of the human population. Its molecular basis is largely unknown, but many loci have been mapped to the X chromosome. We report on two mentally retarded females with X;autosome translocations and breakpoints in Xp11, viz., t(X;17)(p11;p13) and t(X;20)(p11;q13). (Fiber-) FISH analysis assigned the breakpoints to different subbands, Xp11.4 and Xp11.23, separated by approximately 8 Mb. High-resolution mapping of the X- chromosome breakpoints using Southern blot hybridization resulted in the isolation of breakpoint-spanning genomic subclones of 3 kb and 0. 5 kb. The Xp11.4 breakpoint is contained within a single copy sequence, whereas the Xp11.23 breakpoint sequence resembles an L1 repetitive element. Several expressed sequences map close to the breakpoints, but none was found to be inactivated. Therefore, mechanisms other than disruption of X chromosome genes likely cause the phenotypes. PMID- 11060463 TI - Chromosome mapping of the genes for murine arylamine N-acetyltransferases (NATs), enzymes involved in the metabolism of carcinogens: identification of a novel upstream noncoding exon for murine Nat2. AB - Arylamine N-acetyltransferases (NATs) catalyse acetylation reactions which can result in either detoxification or activation of arylamine carcinogens. The human NAT loci (NAT1, NAT2, and a pseudogene, NATP) have been mapped to human chromosome 8p22, a region frequently deleted in tumours. There are three functional genes in mice (Nat1, Nat2, and Nat3) encoding for three NAT isoenzymes. Different alleles at the Nat2 locus are responsible for the acetylation polymorphism identified in different mouse strains. We show that Nat3 is close to Nat1 and Nat2, by screening of a P1 artificial chromosome (PAC) library and provide cytogenetic evidence for co-localisation of the three genes in chromosome region 8 B3.1-B3.3. The Nat region of mouse and human is homologous. We also provide sequence information and a restriction map in the vicinity of Nat1 and Nat2 and describe a noncoding exon located 6 kb upstream of the Nat2 coding region. PMID- 11060464 TI - Organization of telomere sequences in birds: evidence for arrays of extreme length and for in vivo shortening. AB - Telomeres are the specialized ends of chromosomes consisting of highly conserved repeat (5'-TTAGGG-3')(n) sequences. Lack of information regarding the existence of an in vivo telomere clock function in birds, conflicting data regarding telomere array length in the chicken model, and the paucity of molecular telomere information for other avian species led us to study telomere array organization within and among 18 species and subspecies of birds. Most of the species contained between 2% and 4% telomere sequence per diploid genome. Arrays spanning 0.5-10 kb (Class I) and 10-40 kb (Class II) were observed in all of the species studied. Extremely long arrays, ranging from hundreds of kilobases to 1-2 Mb (Class III) were observed in all except two raptor species, the northern goshawk and American bald eagle. In chicken, there was evidence for shortening of the Class II arrays in vivo, based on intraindividual comparisons of somatic versus germline tissues in birds of different ages; terminally differentiated erythrocyte arrays were, on average, 2.3 kb shorter than sperm (germline) arrays. This study provides the first evidence for the existence of telomere arrays significantly larger than have been described for any vertebrate species to date and for developmentally programmed in vivo telomere shortening in the Aves taxa. The novel finding of megabase-sized telomere arrays may be an important feature of avian karyotypes that contain a large number of very small genetic units, the microchromosomes. PMID- 11060465 TI - Cloning and chromosomal mapping of the mouse and human genes encoding the orphan glucocorticoid-induced receptor (GPR83). AB - The mouse glucocorticoid-induced receptor (GIR) is an orphan G protein-coupled receptor highly expressed in brain and thymus (Harrigan et al., 1989; 1991). We have cloned the mouse GIR gene (Gpr83), determined its genomic organization and compared it with the human gene. The genomic organization of the gene is similar in both species although differences leading to specific splicing variants in the mouse have been found. Three introns interrupting the coding sequence are common to both mouse and human. A short sequence in the second intron of the mouse gene can be alternatively spliced in, leading to an insertion in the second intracellular loop of the receptor. This insertion constitutes an additional exon which is not present in the human genome. The human GIR polypeptide shares 89.5% and 91.5% identity with its mouse and dog orthologs respectively. Splice variants lacking the first extracellular loop and the third transmembrane domain have been found in human and mouse species. The receptor variants resulting from these minor transcripts are likely to be non functional. Comparative genetic mapping of the Gpr83 gene showed that it maps to regions of conserved synteny on mouse chromosome 9 (A2-3 region) and human chromosome 11 (q21 region). PMID- 11060466 TI - Clustering of two fragile sites and seven homeobox genes in human chromosome region 2q31-->q32.1. AB - In this study we have used FISH to examine the relationship between a group of homeobox genes, namely DLX1/DLX2, EVX2 and four HOXD genes (10, 11, 12, 13), that map to region q31 on chromosome 2, and the FRA2G and FRA2H fragile sites located at 2q31 and 2q32.1 respectively. Our results indicate that these homeobox genes lie between the two fragile regions. PMID- 11060467 TI - Electrolytic treatment of colorectal liver tumour deposits in a rat model: a technique with potential for patients with unresectable liver tumours. AB - BACKGROUND/AIMS: Patients with unresectable malignant liver tumours have a poor prognosis. A technique is needed which improves long-term survival. Previous studies in the rat have shown that electrolysis is a safe, predictable and reproducible method for creating areas of necrosis in the normal rat liver. This study examined the effects of electrolysis on colorectal liver 'metastases' in the rat. METHODS: Tumours of colorectal origin were implanted into the livers of Wistar-WAG rats. Two weeks after implantation the tumours were treated with electrolysis. A direct current generator, connected to 2 platinum intrahepatic electrodes was used to examine the effects of various electrode configurations on the extent of tumour necrosis. RESULTS: Significant (p<0.001) tumour ablation was achieved with all electrode configurations. Tumour necrosis was more complete (p<0.05) with the electrodes positioned on either side of the tumour than with both electrodes placed in the centre of the tumour. Liver enzymes (AST and ALT) were significantly (p<0.001) elevated after treatment, but returned towards normal by 2 days. CONCLUSIONS: This study has shown that colorectal liver 'metastasis' can be ablated by electrolysis in a rat model. Two separate mechanisms of tumour ablation were observed: With the electrodes directly in or adjacent to the tumour, necrosis resulted from the action of cytotoxic electrode products, whereas by positioning the electrodes proximal to the tumour, necrosis was induced by a 'secondary' ischaemic effect. The findings confirm the view that electrolysis has great potential for treating patients with unresectable malignant liver tumours. PMID- 11060468 TI - New perspectives in gastric acid suppression: genetic polymorphisms predict the efficacy of proton pump inhibitors. AB - Proton pump inhibitors are highly effective in the management of acid-peptic diseases. These drugs potently inhibit acid secretion from gastric parietal cells by irreversibly inhibiting activity of the H(+), K(+) ATPase (proton pump). Early studies of the pharmacokinetics of proton pump inhibitors demonstrated considerable variation in drug clearance rates among patients and healthy volunteers. This variation was also reflected in a wide range of the efficacy of acid suppression by standard doses of proton pump inhibitors among study subjects; those with slower clearance and higher drug concentrations experienced superior acid suppression. Proton pump inhibitors are predominantly inactivated by the 2C19 isoform of the hepatic cytochrome P450 mixed function oxidase system. The cytochrome P450 2C19 gene is polymorphic, with three known inactivating mutations. Individuals with one or two mutant cytochrome P450 2C19 alleles metabolize proton pump inhibitors more slowly than those with two wild-type alleles and experience higher drug levels. An individual's cytochrome P450 2C19 genotype predicts the degree of acid suppression in response to a standard dose of a proton pump inhibitor. Emerging data suggests that the clinical effectiveness of proton pump inhibitors in the treatment of acid-peptic diseases may also be dependent on cytochrome P450 2C19 genotype. PMID- 11060469 TI - Topographic esophageal manometry: an emerging clinical and investigative approach. AB - Topographic esophageal manometry utilizes an increased number of pressure sensors and three-dimensional displays to fully reveal the pressure continuum representing peristalsis. The techniques demonstrate that peristalsis is comprised of a chain of pressure segments, beginning at the upper esophageal sphincter and continuing through lower sphincter aftercontraction. Topographic methods have already proven useful in both research and clinical settings. Description of the techniques, the topographic representation of normal and abnormal esophageal motility, and advantages of topography over conventional manometric methods are discussed in this review. PMID- 11060470 TI - Fecal occult blood testing for iron deficiency: a reappraisal. AB - Fecal occult blood (FOB) tests have been evaluated primarily for the application of colorectal cancer screening. Less is known about the performance characteristics of FOB tests for the evaluation of iron deficiency, the most common other application. As most clinically important occult gastrointestinal bleeding arises from the proximal gut, it is critical that FOB tests target analytes that are stable during the enteric transit. Available data indicate that guaiac-type and immunochemical tests are insensitive for the detection of proximal gut bleeding, and their use may confound the evaluation of iron deficiency. In contrast, the heme porphyrin test is sensitive for both proximal and distal sources of occult gastrointestinal bleeding, and this FOB test would appear to be the most rational selection for use in patients with iron deficiency or anemia. Outcome data are needed to better assess the impact of FOB testing on algorithms for evaluation of iron deficiency. PMID- 11060471 TI - Chronic intestinal pseudo-obstruction. AB - Chronic intestinal pseudo-obstruction (CIPO) is a syndrome defined by the presence of chronic intestinal dilation and dysmotility in the absence of mechanical obstruction or gross inflammatory disease. Specific diseases may affect any level of the brain-gut axis. For most patients, the diagnosis relies upon a combination of historical, laboratory, manometric and histological features. Recent advances into the autoimmune nature of etiologies such as Chagas' disease and paraneoplastic dysmotility and into the genetic basis of mitochondrial neurogastrointestinal encephalomyopathy, multiple endocrine neoplasia IIB and Hirschsprung's disease have greatly refined our understanding and diagnosis of these disorders. At present, medical therapy of CIPO remains limited. Current and future developments in pharmacologic agents targeting specific enteric neurotransmitters and motility patterns hold much promise for improving the care of the patients afflicted with this complex and often debilitating syndrome. PMID- 11060472 TI - Pathophysiology and pharmacological treatment of gastroesophageal reflux disease. AB - Gastroesophageal reflux disease (GERD) is one of the most common diagnoses in a gastroenterologist's practice. Gastroesophageal reflux describes the retrograde movement of gastric contents through the lower esophageal sphincter (LES) to the esophagus. It is a common, normal phenomenon which may occur with or without accompanying symptoms. Symptoms associated with GERD include heartburn, acid regurgitation, noncardiac chest pain, dysphagia, globus pharyngitis, chronic cough, asthma, hoarseness, laryngitis, chronic sinusitis and dental erosions. The introduction of fiberoptic instruments and ambulatory devices for continuous monitoring of esophageal pH (24-hour pH monitoring) has led to great improvement in the ability to diagnose reflux disease and reflux-associated complications. The development of pathological reflux and GERD can be attributed to many factors. Pathophysiology of GERD includes incompetent LES because of a decreased LES pressure, transient lower esophageal sphincter relaxations (TLESRs) and deficient or delayed esophageal acid clearance. Uncomplicated GER may be treated by modification of life style and eating habits in an early stage of GERD. The various agents currently used for treatment of GERD include mucoprotective substances, antacids, H(2) blockers, prokinetics and proton pump inhibitors. Although these drugs are effective, they do not necessarily influence the underlying causes of the disease by improving the esophageal clearance, increasing the LESP or reducing the frequency of TLESRs. The following article gives an overview regarding current concepts of the pathophysiology and pharmacological treatment of GERD. PMID- 11060473 TI - Esophageal cast: a manifestation of graft-versus-host disease. AB - Diffuse involvement of the gastrointestinal tract by graft-versus-host disease (GVHD) is a common complication of bone marrow transplantation. The esophageal involvement in this disease tends to be a vesiculobullous, ulcerative or desquamative process. To our knowledge, esophageal cast has not been described in the context of GVHD. However, it has been described as a result of trauma to the esophagus or in association with bullous disease of the skin. We present a case of esophageal cast in a patient with chronic GVHD following bone marrow transplant. PMID- 11060474 TI - Variation between and within drug treatment modalities: data from the National Treatment Outcome Research Study (UK). AB - This paper describes treatment practices in 54 drug treatment programmes taking part in the National Treatment Outcome Research Study (NTORS). Programmes were representative of the 4 main treatment modalities in the UK: in-patient, residential rehabilitation, methadone maintenance and methadone reduction. Distinguishing features of these forms of treatment were identified. Substantial variations in treatment practices were also observed within each modality, particularly for detoxification and prescribing arrangements, counselling and non drug treatment services, and planned duration of treatment. Many programmes reported extensive waiting lists. These findings are discussed in the context of the growing international evidence of the association between patient outcome and the manner in which programme services are delivered. PMID- 11060475 TI - New European instruments for treatment outcome research: reliability of the maudsley addiction profile and treatment perceptions questionnaire in Italy, Spain and Portugal. AB - This report describes the field testing of two recently developed instruments for treatment evaluation research - the Maudsley Addiction Profile (MAP) and the Treatment Perceptions Questionnaire (TPQ) - in Italy, Portugal and Spain. The MAP and TPQ have been developed in the United Kingdom as brief instruments which contain measures of high face validity for research applications with the adult psychoactive substance use disorder population. The present study assesses the application of these instruments in the continental European context and assesses the internal and test-retest reliabilities of the items. A total of 206 subjects participated in the study (124 subjects participated in the MAP test-retest study, and 95 clients completed the TPQ). Thirteen subjects completed both the MAP and the TPQ questionnaires. Results of the study indicated that the MAP can be administered to clients in 15 min or less. The internal and test-retest reliabilities of the MAP and TPQ are satisfactory. Both instruments are suitable for treatment evaluation and other relevant research purposes in the European Union. PMID- 11060476 TI - Effectiveness of inpatient drug detoxification: links between process and outcome variables. AB - The Psychiatric University Hospital of Zurich offers a 2-week residential detoxification treatment program for patients abusing illegal drugs. 96 out of 160 patients were interviewed at three intervals: upon admission, on the 10th day of treatment, and about 1 month after leaving the clinic. Approximately 17% of the patients were completely abstinent 1 month after treatment. About 50% of the patients could give up the consumption of particular drugs. For around one third of the dependents, the drug consumption remained the same or had increased. The analysis of the predictors of drug consumption after treatment stresses the importance of the social integration of the dependents before detoxification as well as of characteristics of the treatment process (relapse, dropping out of treatment, and referral to a follow-up program). PMID- 11060477 TI - An open study with acamprosate in Belgium and Luxemburg: results on sociodemographics, supportive treatment and outcome. AB - This open study on 614 alcohol-dependent patients from 51 centres in Belgium over 24 weeks collected data on the sociodemographic, medical and drug safety profiles and the type of psychosocial support most commonly associated with acamprosate. Psychiatric problems (34%) and gastro-intestinal ulcers (15%) were the most frequent other medical diagnoses. As supportive treatment, 48% of patients received individual psychotherapy, 25% group therapy, 16% relapse prevention and coping skills, 9% brief intervention and 2% family therapy. Fifteen (29%) centres used only 1 form of supportive treatment, 14 (27%) used 2 forms of support, 16 (31%) used 3 forms, 4 (8%) used 4 and only 2 centres (4%) used all 5 supportive treatment options. Three hundred and fifty-nine patients dropped out of the study. At any moment during the treatment period, up to 11% relapsed, 9% had binge drinking and 9% had drinking lapses. The quantity of alcohol consumption during relapse, lapse or binge drinking showed a considerable drop in the mean number of drinks per day, from 19.53 before treatment to 4.23-7.83 drinks for lapse drinking, 9.27-14.62 drinks for binge drinking and 4.96-10.29 drinks per day for relapse. All 8 dimensions of the SF36 quality of life questionnaire improved over the treatment period. Acamprosate was well tolerated by all patients. PMID- 11060478 TI - Pathways to abstinence: two-year follow-up data on 60 abstinent former opiate addicts who had been turned away from treatment. AB - Structural changes in the organization of drug treatment services in the northwest of England during the early 80s provided a unique opportunity to study a group of opiate addicts who were turned away from treatment. This paper reports on 60 opiate addicts who were abstinent at follow-up (2-3 years after their original referral). For the majority of subjects, any treatment received in the intervening period had been from a range of local, non-specialist treatment providers, including: general practices (n = 24) and local hospitals (n = 32). The subjects had also received assistance from non-statutory agencies (n = 35). Informal sources of support were drawn upon in the form of friends, family, and voluntary agencies. Social acceptance, legal problems, financial difficulties, and imprisonment were frequently cited as motivational factors that led to abstinence. At follow-up, improvements were reported in personal relationships and family circumstances, although there was less progress in relation to employment, finances, and housing. Increased involvement with the criminal justice system was significantly associated with a greater use of treatment services at follow-up. This paper presents the treatment and non-treatment pathways that led to abstinence amongst this group of opiate addicts. It also points to the importance of including non-treatment samples in evaluations of treatment interventions. PMID- 11060479 TI - Criminality in female inpatients with substance use disorders. AB - Three groups of female inpatients were compared with each other with regard to criminal behavior: 51 patients with alcohol dependence, 39 patients with drug (mostly opioid) dependence and 488 patients not fulfilling DSM-IV criteria for substance dependence. 45% of the alcohol-dependent patients, 74% of the drug dependent patients and 15% of the psychiatric comparison group had a criminal record compared to only 2% of the female reference general population. Comparing criminal with noncriminal patients in both groups with substance dependence, patients with a criminal record were significantly younger than patients without a criminal record; however, otherwise only very few differences appeared. In contrast, alcohol-dependent and drug-dependent female patients with a criminal record appear to represent two quite different populations. PMID- 11060480 TI - A comparison of heroin chasers with heroin injectors in Switzerland. AB - In a cross-sectional multicentre study, we compared the characteristics of heroin chasers and heroin injectors. Subjects were 162 primarily opioid-dependent volunteers for whom either chasing (n = 85) or injecting (n = 77) was the principal route of heroin administration. Each subject was rated by means of the Swiss version of the European Addiction Severity Index. Additionally, subjects completed a questionnaire battery including the Severity of Dependence Scale, the Symptom Checklist SCL-90-R, a self-constructed peergroup questionnaire and a semantic differential list to assess the connotative meaning of heroin chasers and injectors. The heroin injectors were older and more likely to use one or more other drugs besides heroin than the chasers. They had longer heroin-using careers, a longer duration of detention and a higher prevalence of some type of hepatitis. In conclusion, the differences between chasers and injectors were rather related to a longer history of heroin use than to the route of heroin administration. PMID- 11060481 TI - Structure and biology of stinging insect venom allergens. AB - Bees, fire ants and vespids cause insect sting allergy. These insects have unique as well as common venom allergens. Vespids, including hornets, paper wasps and yellow jackets, have common allergens. Bees and vespids have one common allergen with hyaluronidase activity; they also have unique allergens with different phospholipase activities. Fire ants and vespids have one common allergen, antigen 5 of unknown biologic activity. The common venom allergens with < 70% sequence identity have barely detectable levels of antigenic cross-reactivity. Possible uses of modified allergens for immunotherapy are described. PMID- 11060482 TI - The Th2 cytokine environment of the placenta. AB - It is now accepted that local changes to the balance of Th1/Th2-type cytokines occur during pregnancy within the maternal uterus and fetoplacental unit. These changes in cytokine profiles contribute to implantation of the embryo, development of the placenta, and survival of the fetus to term. Overall within the placenta there is a bias in the ratio of Th1:Th2 cytokines towards the Th2 type cytokines. However, there are specific fluctuations in this balance at implantation and during the initiation of parturition. The predominant cytokines at each stage of gestation function both to limit maternal immune rejection of the semi-allogeneic embryo/fetus, especially at the maternofetal interface; and to facilitate the on-going physiological processes within the maternal reproductive tract. These two, at times conflicting, roles are discussed in this review, with key evidence concerning cytokine expression and function from mouse and humans. PMID- 11060483 TI - Molecular cloning of paramyosin, a new allergen of Anisakis simplex. AB - BACKGROUND: Anisakis simplex is a fish parasite that, when accidentally ingested by humans, may cause allergic reactions in sensitized individuals. The main objectives of our study were to: (1) construct a cDNA expression library of A. simplex; (2) identify clones producing specific IgE binding protein antigens, and (3) produce and purify the protein/s codified by the isolated clones produced in Escherichia coli. METHODS: An expression cDNA library from the third stage larvae (L3) of A. simplex was constructed. This library was first screened with a rabbit anti A. simplex hyperimmune serum. The positive clones, identified using the rabbit serum, were rescreened with a pool of human sera containing high titers of IgE antibodies against A. simplex. RESULTS: Two positive clones were isolated carrying the genes which codify for paramyosin. The paramyosin protein was produced in E. coli and purified. The partial sequence of a second paramyosin gene was also identified. The frequency of specific IgE binding to the recombinant and native forms of paramyosin using the sera of 26 A. simplex sensitive individuals was 23 and 88%, respectively. Both paramyosins were able to inhibit 11% of the specific IgE binding to a total extract. CONCLUSIONS: We describe the primary structure of a paramyosin of A. simplex. It can be considered as an allergen based on its IgE binding capacity. We suggest that the recombinant protein does not maintain the complete allergenic properties of the native paramyosin, considering its lower IgE binding capacity of the recombinant protein. However, both proteins have the same specific IgE inhibition capacity. The recombinant protein can be produced in large quantities in E. coli. We propose the term Ani s 2 for this allergen. PMID- 11060484 TI - Electrophoretic and immunochemical characterization of allergenic proteins in buckwheat. AB - BACKGROUND: Buckwheat allergies are not common, however, it is considered to be a very potent allergen. Ingestion of small amounts has been found to produce anaphylactic reactions, particularly in children. Identification and characterization of the major allergen(s) in buckwheat are currently underway, however, there are some discrepancies in the findings. METHODS: Identification of the major allergen(s) was determined through Western blotting using buckwheat allergic patients' sera. Once the allergenic proteins were identified, they were purified, their IgE-binding activity assessed through an indirect ELISA and the N terminal amino acid sequence completed. To assess the stability of the IgE binding epitopes, protein fractions were exposed to various treatments and assayed using an indirect ELISA. Lastly, the presence of anti-buckwheat IgG in the patients' sera was analyzed through Western blotting and ELISA. RESULTS: IgE binding was detected to proteins with molecular masses of approximately 14 and 18 kDa. N-terminal sequencing was completed and found to share some homology with rice proteins associated with rice allergies and cross-allergenicity with buckwheat proteins. When the water-soluble protein fraction was heated, exposed to acidic and alkaline conditions and fully denatured, IgE-binding activity was reduced. When the fraction was partially denatured through urea, IgE-binding activity increased. Furthermore, IgG-binding activity was detected with proteins only above the 20 kDa region. CONCLUSIONS: Proteins with molecular masses around 14 and 18 kDa were identified as the major allergenic proteins in the buckwheat allergic patients' sera tested in this study. Results also indicate that these two proteins possess IgE-binding capability. PMID- 11060485 TI - Cultivar-specific IgE-epitopes in date (Phoenix dactylifera L.) fruit allergy. Correlation of skin test reactivity and ige-binding properties in selecting date cultivars for allergen standardization. AB - BACKGROUND: Date fruits are allergenic and standardized extracts are required for diagnosis and therapy of this allergy. Since there are several cultivars of dates, this study was carried out to assess the allergenicity of different cultivars in order to select suitable source material for standardization. METHODS: The protein profiles of 18 of the most commonly sold varieties were compared by SDS-PAGE and their relative allergenicity assessed by SPT and IgE based ELISA and immunoblotting. Thirty-two date fruit-sensitive patients were skin tested with a pooled extract from all the cultivars. Six of the patients with high SPT results (> or =3+) who volunteered were further tested with the 18 cultivars and their sera used in ELISA and immunoblotting. RESULTS: Six of the cultivars gave high SPT-positive reactions in > or =4 of patients. Five of these high SPT-reactive cultivars gave high IgE ELISA scores (> or =0.58) but individual cultivars varied in their number of IgE immunoblot bands. Cultivar specific IgE-binding patterns indicated that only certain cultivars bound IgE at molecular weights of < or =14.3 and 27-33 kDa whilst all cultivars bound to a 54 58 kDa doublet. Cultivars that bind to the < or =14.3 and 27-33 kDa bands appeared to form the majority of the high SPT-reactive cultivars. When individual sera of 24 of the 32 SPT-positive patients were used in IgE immunoblots with the pooled cultivar extract, all sera bound IgE at < or =14.3 and 27-33 kDa and about 60% of sera bound to a 54-58 kDa doublet bands. CONCLUSIONS: These results indicate that allergenicity of date fruits is a cultivar-specific phenomenon. Sixty to 100% of sera from date fruit-allergic patients bind IgE to three major allergens of < or =14.3, 27-33 and 54-58 kDa. Five of the cultivars that evoke high SPT reactions, high IgE ELISA scores and bind IgE to the major allergens, can be selected for the preparation of 'in-house' allergen extracts and for allergen standardization. PMID- 11060486 TI - Inverse association between skin response to aeroallergens and Schistosoma mansoni infection. AB - BACKGROUND: Helminthic infections and allergic disease are highly prevalent in many areas of the world. It is known that IgE antibodies are involved in the pathogenesis of both helminthiasis and atopy. However, the consequences of the presence of helminthic infections in atopic patients are still not completely understood. METHODS: Subjects infected by Schistosoma mansoni with more than 200 eggs/g of feces (n = 42) and uninfected subjects (n = 133) were selected from an endemic area of schistosomiasis. The history of allergy and results of the immediate hypersensitivity prick tests with inhalant allergen extracts were registered. Total IgE and IgE specific to S. mansoni and aeroallergens were measured in serum by ELISA. RESULTS: The proportion of individuals with a positive skin test to allergens was higher in the uninfected group (24.3%) than in the group with more than 200 eggs/g of feces (4.8%). The odds of atopy (defined as a positive test for at least one of the antigens) were 5 times higher (odds ratio = 7.0; 95% confidence interval = 1.6-31.1%; p = 0.01) in the uninfected group, after taking into account the potential influence of gender and age. While there was a tendency for higher total and S. mansoni-specific IgE levels in infected patients, an opposite trend, that is higher aeroallergen specific IgE, was observed in uninfected subjects. CONCLUSIONS: There was a strong and statistically significant inverse association between the immediate skin test response to common aeroallergens and infection by S. mansoni. The results indicate that immediate hypersensitivity reactions may be suppressed in S. mansoni-infected individuals. PMID- 11060487 TI - Anti-tetanus toxoid antibody production and protection against lethal doses of tetanus toxin in hu-PBL-SCID mice. AB - BACKGROUND: In this study, severe combined immunodeficiency (SCID) mice, which permit the survival of lymphoid cells of human origin, were used to study the human anti-tetanus immune response. METHODS: Human peripheral blood lymphocytes (hu-PBL) obtained from 88 healthy donors (aged from 18 to 62) were transplanted into SCID mice, and anti-tetanus toxoid (Ttd) antibody production and protection against lethal doses of tetanus toxin (Ttx) were investigated in the hu-PBL-SCID mice. RESULTS: The transfer of human PBL evoked significant human anti-Ttd IgG antibody production for 37.5% of the donors. After in vivo immunization, the percentage of donors with PBL exhibiting positive anti-TtD IgG production in the mice increased to 54.5%. Mean anti-Ttd IgG levels in the sera were also significantly elevated in response to immunization. The mean IgG titer for the mice injected with PBL from donors under the age of 40 was significantly higher than that of the mice injected with PBL from donors aged 40 or older. Four weeks after the cell transfer, the mice were challenged with Ttx. The induction of protection against Ttx challenge was observed mostly in mice with PBL transferred from donors under the age of 40. In vivo immunization in SCID mice with Ttd increased the number of cases of resistance to Ttx. CONCLUSIONS: These results suggest that hu-PBL-SCID mice might serve as a tool for predicting the protective ability against pathogens in PBL donors and also for evaluating vaccine efficacy. PMID- 11060488 TI - Hexamethylene diisocyanate causes contraction of canine tracheal smooth muscles through activation of muscarinic receptors. AB - BACKGROUND: Asthma caused by occupational exposure to hexamethylene diisocyanate (HDI) is well known; however, the exact pathogenic mechanisms remain unclear. METHODS: Experiments were performed using a standard canine tracheal smooth muscle (CTSM) strip preparation in an isolated bath to determine the effect of HDI on tracheal smooth muscle contraction. HDI concentration-response curves were constructed and the effects of different receptor antagonists on HDI-induced smooth muscle contraction were determined. To determine whether HDI and acetylcholine (ACh) bind to a common muscarinic receptor, ACh concentration response curves in the absence or presence of HDI and concentration-response curves for HDI and ACh in the presence or absence of atropine were plotted. RESULTS: HDI caused contraction of CTSM, with a threshold concentration of 10(-7) M. The EC(50) (HDI concentration that produced 50% of the maximal response) was 6.2+/-0.7 x10(-7) M and the maximal contractile response (174+/-55 g/g of tissue) occurred at a concentration of 5.0+/-0.8 x 10(-6) M. Atropine, a muscarinic blocker, significantly inhibited HDI-induced contractile responses. HDI shifted the ACh concentration-response curve to the right. The mean pA(2) for atropine against ACh (8.93+/-0.27) was not significantly different from that against HDI (8.03+/-0.12). CONCLUSIONS: Our results indicated that HDI causes contraction of CTSM through the activation of muscarinic receptors. Direct stimulation of muscarinic receptors by HDI may play an important role in HDI-induced asthma. PMID- 11060489 TI - Effects of mizolastine in vitro on human immunocompetent and airway cells: evidence for safety and additional property. AB - BACKGROUND: Mizolastine is a potent, peripherally acting, selective H1-receptor antagonist with potential anti-inflammatory properties. The aim of the study was to evaluate the in vitro effects of mizolastine on the expression of adhesion molecules by primary human airway epithelial and stromal cultures; moreover, the activity of mizolastine on parameters which reflect the immune response efficacy was investigated. METHODS: Airway epithelial and stromal cells were collected from hypereosinophilic subjects by enzymatic digestion of polyps or turbinates. Cells were stimulated with interferon (IFN)-gamma (500 IU/ml) in the presence of various mizolastine concentrations (6 x 10(-8)-6 x 10(-6) M) for 24 h and the expression of CD106, CD54, CD58 and HLA class I was evaluated. Peripheral blood mononuclear cells from healthy volunteers were incubated with 1% phytohemagglutinin or anti-CD3 monoclonal antibody (20 ng/ml) in the presence of mizolastine, then T lymphocyte proliferation, HLA-DR expression and T cell subpopulations were evaluated. RESULTS: Both in epithelial and stromal cultures, IFN-gamma significantly upregulated all of the tested surface molecules (p<0.05). The highest dose of mizolastine (6 x 10(-6) M), corresponding to 10-fold the peak plasma level after a single oral administration of 10 mg, was able to act on fibroblasts, significantly downregulating the expression of CD54 (p<0.05). Regarding T lymphocyte proliferation, the addition of mizolastine did not induce any significant change; furthermore, mizolastine was ineffective at all of the tested concentrations on both HLA-DR expression and CD4+/CD8+ ratio. CONCLUSIONS: This study demonstrated that mizolastine is able to selectively downregulate CD54 expression on stimulated stromal but not epithelial cells without impairing the immune system effectors. The possible clinical significance of these results are an antiallergic property and CD54 modulation on fibroblasts with a good safety profile as far as the lymphocyte response is concerned. PMID- 11060490 TI - Early DNA synthesis and cytokine expression in the nickel activation of peripheral blood mononuclear cells in nickel-allergic subjects. AB - BACKGROUND AND OBJECTIVE: Nickel sulphate stimulates the proliferation of lymphocytes in nickel-allergic subjects. However, nickel-induced stimulation of lymphocytes from control persons without clinical symptoms of nickel allergy has also been reported. The aim of the present study was to correlate T cell activity, measured by DNA synthesis and the expression of Th1 [interleukin (IL) 2, tumour necrosis factor (TNF)-beta and interferon (IFN)-gamma] and Th2 (IL-4) cytokines, in short-term (up to 72 h) culture of nickel-stimulated peripheral blood mononuclear cells (PBMC) from nickel-allergic patients compared to control subjects. METHODS: DNA synthesis was measured by the incorporation of [methyl (3)H]thymidine. The production of IL-2, TNF-beta, IFN-gamma and IL-4 was measured in the supernatants of the cultures by ELISA. In situ hybridization for mRNA was performed using oligonucleotide probes for IL-4, IFN-gamma and TNF-beta in cell smears. RESULTS: Already after 24 h and proceeding through the remaining culture period, there was a statistically significant (p<0.001) difference in the concentrations of IL-2 between patients and controls. There was a significant (p<0.01) difference in DNA synthesis (stimulation index) between the patients and control subjects at 72 h, and also at the same time a difference in the concentrations of TNF-beta (p<0.05) and IL-4 (p<0.01). In the in situ hybridization study, TNF-beta was found to be the only one of the studied cytokines that differed between the nickel-allergic and control subjects, this difference being most evident at 72 h (p<0.01). CONCLUSIONS: Our results indicate a difference between nickel-allergic and non-allergic subjects in the synthesis of DNA and production of cytokines when PBMC are stimulated by nickel sulphate, and IL-2 may be regarded as a critical and early-occurring cytokine. PMID- 11060491 TI - IgA deficiency in Czech healthy individuals and selected patient groups. AB - BACKGROUND: Selective IgA deficiency (IgAD) is the most common immunoglobulin deficiency with a variety of clinical manifestations. The frequency of IgAD differs depending on the ethnic origin and clinical symptoms of investigated persons. METHODS: The prevalence of IgAD (serum IgA level <0.05 g/l) was determined in 5,310 Czech blood donors, 10,326 patients who had undergone immunological investigation, and 246 first-degree relatives of IgAD and common variable immunodeficiency (CVID) patients. RESULTS: IgAD was detected in 13 (1/408; 0.24%) of the blood donors. The prevalence of IgAD was increased both in children (48/3,113; 1.5%) and adults (33/3,824; 0.9%) referred for frequent respiratory tract infections (in both cases p<0.001) compared to the healthy population. The frequency of IgAD was 12/189 (6%) in first-degree relatives of IgAD patients and 9/57 (16%) in relatives of CVID patients, with the highest frequency observed in children of CVID patients. CONCLUSIONS: The prevalence of IgAD in the Czech healthy population is comparable to that in other Caucasians. The frequency is increased in children with recurrent respiratory tract infections and especially in relatives of patients with immunoglobulin deficiencies. PMID- 11060492 TI - Novel signaling pathways contributing to vascular changes in hypertension. AB - In hypertension, increased peripheral resistance maintains elevated levels of arterial blood pressure. The increase in peripheral resistance results, in part, from abnormal constrictor and dilator responses and vascular remodeling. In this review, we consider four cellular signaling pathways as possible explanations for these abnormal vascular responses: (1) augmented signaling via the epidermal growth factor receptor to cause remodeling of the cerebrovasculature; (2) reduced sphingolipid signaling leading to blunted vasodilation and increased smooth muscle proliferation; (3) increased signaling via Rho/Rho kinase leading to enhanced vasoconstriction, and (4) a relative state of microtubular depolymerization favoring vasoconstriction in hypertension. These novel cell signaling pathways provide new pharmacological targets to reduce total peripheral vascular resistance in hypertension. PMID- 11060493 TI - Actions of melatonin in the reduction of oxidative stress. A review. AB - Melatonin was discovered to be a direct free radical scavenger less than 10 years ago. Besides its ability to directly neutralize a number of free radicals and reactive oxygen and nitrogen species, it stimulates several antioxidative enzymes which increase its efficiency as an antioxidant. In terms of direct free radical scavenging, melatonin interacts with the highly toxic hydroxyl radical with a rate constant equivalent to that of other highly efficient hydroxyl radical scavengers. Additionally, melatonin reportedly neutralizes hydrogen peroxide, singlet oxygen, peroxynitrite anion, nitric oxide and hypochlorous acid. The following antioxidative enzymes are also stimulated by melatonin: superoxide dismutase, glutathione peroxidase and glutathione reductase. Melatonin has been widely used as a protective agent against a wide variety of processes and agents that damage tissues via free radical mechanisms. PMID- 11060494 TI - Apoptosis occurs in a new model of thermal brain injury. AB - Apoptosis has been implicated recently as a prominent response of the brain to a variety of insults, such as ischemia and trauma. In this study, we demonstrate that apoptosis is a prominent part of the brain's response to a thermal insult. To examine the brain's response to a thermal insult, a new model of thermal brain injury in the laboratory rat was developed. Water heated to 60 degrees C was passed over an area of thinned calvarium for 1 min. This resulted in an actual brain temperature of 47-48 degrees C. A uniform area of 2,3,5-triphenyl tetrazolium chloride pallor was demonstrated and pyknotic neurons were seen in the area of injury by hematoxylin-eosin staining. Apoptosis was demonstrated by the characteristic DNA fragmentation seen by agarose gel electrophoresis, ApopTag in situ staining and electron microscopy. The findings of apoptosis were localized to the area of thermal injury and were time dependent, starting 6 h after the insult and peaking approximately 18 h after the insult. This represents one of the first demonstrations that apoptosis occurs in the brain in response to a thermal injury. PMID- 11060495 TI - Age-associated changes of superoxide dismutase and catalase activities in the rat brain. AB - Oxygen free radicals have been proposed to be involved in the process of aging. Superoxide dismutase (SOD) and catalase are important for antioxidative defense. In this study, profiles of SOD, catalase, and their mRNA levels were investigated in the frontal, parietal, temporal and occipital lobes, subcortex and cerebellum of male Wistar rats at ages 1-21 months. The total SOD and Mn SOD activities increased with age and exhibited higher levels at 6 and 12 months but decreased thereafter. Activity of catalase showed a similar trend and notably peaked at 12 months. The mRNA levels of Cu/Zn SOD, Mn SOD, and catalase remained constant in all areas tested (frontal, parietal, temporal and occipital lobes, and subcortex) except the cerebellum. Post-transcriptional regulation was involved in modulating the enzymes' activities during aging. Furthermore, the rate of mitochondrial generation of the superoxide anion (O(2)(-).) increased gradually with aging. Taken together, the results suggest that the increase of oxidative potential and the loss of proper antioxidant defense in the rats appear to be highly involved in the aging process of the brain. PMID- 11060496 TI - Role of flhDC in the expression of the nuclease gene nucA, cell division and flagellar synthesis in Serratia marcescens. AB - We investigated in Serratia marcescens the functions of the flhDC operon, which controls motility and cell division in enteric bacteria. Included in our evaluations were investigation of cell division, flagellar synthesis and regulation of the expression of nuclease (encoded by the nucA(Sm) gene, one of the virulence factors). Interruption of the chromosomal flhDC operon in S. marcescens CH-1 resulted in aberrant cell division and loss of nuclease and flagella. Expression of nucA(Sm) and other mutated phenotypes was restored in the flhDC mutant by the induction of overexpression of flhDC in a multicopy plasmid. Multicopied flhDC also induced the formation of differentiated cells (polyploid aseptate cells with oversynthesis of peritrichous flagella) in broth culture using minimal growth medium. Expression of the flhDC operon showed positive autoregulation, and was growth phase dependent (upregulated in early log phase). In addition, flhDC expression was inhibited when the temperature increased from 30 to 37 degrees C, and when osmolarity was increased, but was not influenced by glucose catabolite repression. These results show that FlhD/FlhC is a multifunctional transcriptional activator involved in the process of cell differentiation, swarming and virulence factor expression. PMID- 11060497 TI - Functional implication of human serine/threonine kinase, hAIK, in cell cycle progression. AB - Protein phosphorylation is involved in many biological activities and plays important roles in cell cycle progression. In the present study, we identified a serine/threonine kinase, hAIK, from human hepatic cells using degenerated polymerase chain reactions with a pair of primers derived from the highly conserved sequence in the catalytic domain of kinases. The full-length hAIK cDNA was then obtained, which contained 403 amino acids and was homologous to Drosophila Aurora2 and yeast Ipl1 proteins. Northern blotting analysis revealed that hAIK was highly expressed in the testis but not in other tissues. Expressions of hAIK drastically increased in cancer tissues/cell lines but not in fibroblasts or nontumorigenic cell lines. The recombinant hAIK protein phosphorylated itself and histone H1; this phosphorylation activity was totally abolished after a point mutation at the catalytic domain (hAIKm). During the interphase cell, hAIK was found mainly in the cytoplasm; during mitosis hAIK accumulated at the centrosomes. In addition, overexpression of hAIK in cancer cell lines (HEK293T and HeLa) appeared to inhibit cell cycle progression. None of these phenomena were observed in hAIKm whose kinase activity was rendered inactive. Our results suggest that hAIK protein/activity might modulate cell cycle progression by interacting with the centrosomes and/or proteins associated with these structures. PMID- 11060498 TI - Characterization of TRBP1 and TRBP2. Stable stem-loop structure at the 5' end of TRBP2 mRNA resembles HIV-1 TAR and is not found in its processed pseudogene. AB - TRBP1 and TRBP2 cDNAs have been isolated based on the ability of the protein that they encode to bind HIV-1 TAR RNA. The two cDNAs have different 5' end-termini resulting in 21 additional amino acids for TRBP2 protein compared to TRBP1. The corresponding gene is conserved in mammalian species. By PCR amplification of a human library, we have isolated an additional 22 nucleotides in the 5' end of TRBP2 cDNA. Based on the addition of these 22 new nucleotides, the first 87 nucleotides of TRBP2 mRNA can fold into a stable stem-loop structure that resembles TAR RNA. We have also isolated the DNA sequence that represents the TRBP processed pseudogene. The absence of full alignment between TRBP2 full length cDNA and this sequence suggests that the stem-loop structure could have prevented a complete reverse transcription during pseudogene formation. Using different antibodies, three forms of TRBP can be identified in primate cells at 40, 43 and 50 kD, suggesting a differential expression from the cDNAs and post translational modifications. Both TRBP1 and TRBP2 activate the basal and the Tat activated level of the HIV-1 LTR in human and murine cells. Our data indicate that TRBP proteins act at a level prior to Tat function. TRBP could contribute to improved HIV expression in murine models. PMID- 11060499 TI - Mutation L210W of HIV-1 reverse transcriptase in patients receiving combination therapy. Incidence, association with other mutations, and effects on the structure of mutated reverse transcriptase. AB - Mutation L210W of HIV-1 reverse transcriptase (RT) is one of the six main mutations that confer in vivo resistance to zidovudine. Surprisingly, this mutation has received scant appraisal and its contribution to the genotypic resistance to nucleoside analogs is not well understood. The aim of this study was: (1) to study the frequency of mutation L210W in a large collection of HIV-1 sequences (2,049 samples, including 395 DNA and 1,654 RNA sequences) from patients receiving combination therapy, and (2) to analyze its association with the other mutations that confer resistance to zidovudine. A mutation at codon 210 (mainly L210W) was found in 647 (32%) of the 2,049 sequences analyzed. Only 43 (<7%) of these 647 genomes were also mutated at codon 70 (p < 10(-5)). In contrast, 98% of these 647 sequences were also mutated at codon 215 (essentially T215Y/F), and 94% at codon 41 (mainly M41L). These data showing a close association between L210W, T215Y/F, and M41L, and a mutual exclusion between K70R and L210W, were confirmed by analyzing the sequences stored in the HIV-1 sequences available through the Stanford HIV RT and Protease Database. Follow-up studies demonstrated that L210W appeared always after T215Y/F. This observation is consistent with crystallographic studies which suggested that the aromatic side chain of Trp 210 could stabilize the interaction of Phe/Tyr215 with the dNTP binding pocket. This molecular cross-talk between amino acid chains occurs nearby the conserved Asp113 residue. Since the lateral chain of Arg70 may also interact with Asp113, this is likely to create a sterical hindrance around this residue. Thus, the R-->K reversion of codon 70 may represent a compensatory mechanism allowing a functional rearrangement of the dNTP-binding pocket in the mutated RT. PMID- 11060500 TI - Expression of a truncated retroviral envelope gene enhances expression of normal cellular phenotypes. AB - The envelope gene of Moloney murine leukemia virus (Mo-MLV) and its various functional domains have been studied extensively but not as much in terms of their biological effects on cell growth. In this study, we report the biological characterization of a truncated Mo-MLV envelope gene, LN11, which is devoid of a signal peptide. Its expression in various cell types, as compared to the control, enabled the transduced cells to assume a more normal phenotype, which is defined by an increase in contact inhibition and factor dependence, as well as reduced tumorigenicity. LN11-transduced fibroblasts exhibited a higher degree of contact inhibition, assumed a more flattened morphology and were more adherent compared to the control. In v-abl transformed hematopoietic cells, expression of LN11 resulted in slower cell growth, which was due to an enhanced dependence on exogenous growth factors. Enforced expression of LN11 also resulted in a slower rate of tumor development and a reduced tumor load. Thus, modification of a retroviral genome could have a significant impact on cell growth and development. This is one example where we need to consider the safety issue carefully when constructing retrovirus vectors for gene therapy. PMID- 11060501 TI - Neutralizing antibody provided protection against enterovirus type 71 lethal challenge in neonatal mice. AB - Experimental infection with enterovirus type 71 (EV71) induced death in neonatal mice in an age- and dose-dependent manner. The mortality rate was 100% following intraperitoneal inoculation 1-day-old ICR mice and this gradually decreased as the age at the time of inoculation increased (60% in 3-day-old mice and no deaths occurred in mice older than 6 days of age). A lethal dose greater than 10(8) PFU was necessary. Lethargy, failure to gain weight, rear limb tremors and paralysis were observed in the infected mice before death. EV71 was isolated from various tissues of the dead mice. Using a reverse transcription polymerase chain reaction technique with a specific primer pair, a 332-bp product was detected in the tissues that produced a culture positive for EV71. Protection against EV71 challenge in neonatal mice was demonstrated following passive transfer of serum from actively immunized adult mice 1 day after inoculation with the virus. Pups from hyperimmune dams were resistant to EV71 challenge. Additionally, maternal immunization with a formalin-inactivated whole-virus vaccine prolonged the survival of pups after EV71 lethal challenge. PMID- 11060502 TI - Presence of cloning vector sequences in the untranslated region of some genes in Genbank. AB - We found multiple cloning site sequences in the reported untranslated regions (UTR) of several genes in Genbank. The erroneous information can result in the failure to amplify DNA fragments containing untranslated regions by RT-PCR. It is suggested that a BLAST search be performed when primers are designed for PCR amplification of the 5' or 3' UTR of genes to ensure that the reported UTR does not contain plasmid-derived sequences. PMID- 11060503 TI - Executive cognitive impairment: a novel perspective on dementia. AB - In 1994 the American Psychiatric Association added impairment of executive control functions (ECF) to its list of cognitive domains that should be considered in the assessment of dementia. This recommendation has not been widely implemented. None the less, there is growing evidence that ECF impairment is common, strongly associated with disability and functional decline, and not well detected by traditional dementia screening tests. This article reviews the implications of ECF for the epidemiology of dementia. The total number of dementia cases may be much greater than previously thought and we are likely to be selectively missing cases with reversible causes of ECF impairment. PMID- 11060504 TI - Epidemiology of primary central nervous system tumors in Estonia. AB - During the period from 1986 to 1996, 1,665 cases of primary central nervous system (CNS) tumors were identified in the resident population of Estonia. Histological verification was available in 81% of the cases. Gliomas were more common in men, while meningiomas and neurinomas were more common in women. No significant difference was observed between the sexes for all primary CNS tumors. The age-specific incidence increased from the age of 30, reached a maximum in the age range of 50-69 years and declined in the elderly which may reflect under diagnosis. The age-adjusted incidence rate for CNS tumors was 8.5/100,000 population. A comparison of our results with those of a previous study carried out in Estonia revealed a significant histology-specific increase in incidence in all age groups. PMID- 11060505 TI - Cognitive test performance and presence of subclinical cardiovascular disease in the cardiovascular health study. AB - The purpose of the present study was to investigate the relationship between performance on a comprehensive battery of neuropsychological tests and the presence of clinical, subclinical or no cardiovascular disease in an elderly community-dwelling population. The results confirm previous reports of significant associations of age, education and gender with test performance. When performance was examined controlling for these variables, significant associations of disease group were seen with five measures emphasizing speed of performance; Parts A and B of the Trail Making Test, the WAIS-R Digit Symbol and Block Design subtests and category verbal fluency. These results add to the evidence that, in addition to other health implications, cardiovascular disease is related to cognitive functioning in the elderly even at subclinical levels. PMID- 11060506 TI - Relationships between cerebrovascular events, APOE polymorphism and Alzheimer's disease in a community sample. AB - To investigate whether the APOE*4 allele modified the relationship between cerebrovascular events and Alzheimer's disease (AD), we collected evidence of previous stroke or transient ischemic attack (TIA) and determined APOE genotype among 102 subjects with AD and 375 nondemented subjects in a community-based study of dementia. Subjects with a history of stroke or TIA were twice as likely to have AD as subjects without such a history. However, APOE*4 carriers with a history of stroke/TIA were 5 times more likely than APOE*4 carriers without such a history to have AD (odds ratio = 5.3, 95% confidence interval = 1.4-20.9). History of stroke/TIA had little effect on the likelihood of having AD in subjects without an APOE*4 allele. PMID- 11060507 TI - Apolipoprotein E phenotype alone does not influence survival in Alzheimer's disease: a population-based longitudinal study. AB - Apolipoprotein E4 (ApoE4) phenotype is a known risk factor for development of Alzheimer's disease (AD). Contradictory results exist concerning the role of ApoE4 in the rate of decline and mortality in AD. Conflicting findings have also been reported about ApoE and gender interactions with respect to survival. We examined the survival of subjects with AD and non-AD controls with respect to ApoE phenotype and gender in a population-based longitudinal study. Cognitive evaluation was performed for a total of 980 subjects (then aged 69-78 years), and 48 cases with AD were identified. ApoE4 phenotype was more frequently present among subjects with AD. In the whole study population, survival was not related to the presence of AD or ApoE4 phenotype. Risk of death was increased for men compared to women, independently of the ApoE4 phenotype (HR 0.5, 95% confidence interval 0.44-0.69). In subjects with AD, the presence of ApoE4 alone did not influence survival. However, in the AD group, ApoE4-negative men had significantly increased risk of mortality compared to the risk in ApoE4-negative women (p < 0.01). We conclude that the presence of ApoE4 phenotype or AD did not influence mortality in the aged population. Once AD had become manifest, ApoE4 alone did not relate to survival. However, in subjects with AD not carrying ApoE4, men had reduced survival compared to women. PMID- 11060508 TI - Parkinson's disease and environmental factors. Matched case-control study in the Limousin region, France. AB - The study included 140 patients with Parkinson's disease (PD) and 280 non Parkinson age-matched controls to evaluate environmental risk factors associated with PD. The effect of exposure to environmental and dietary factors was determined using conditional logistic regression. This multivariate analysis showed that PD in first-degree relatives and tea drinking were the main risk factors for PD. Smoking appeared to be a protective factor. Exposure to toxic compounds was not a significant risk factor. Further research is needed to validate that tea consumption increases the risk of PD. PMID- 11060509 TI - Epidemiology of Reye's syndrome, United States, 1991-1994: comparison of CDC surveillance and hospital admission data. AB - This investigation describes the epidemiology of Reye's syndrome (RS) during 1991 1994 and compares two different sources of information in the United States. Estimates of the incidence of RS from the Centers for Disease Control and Prevention (CDC) are compared with hospital inpatient data from approximately one third of the hospitals from HCIA, Inc. During 1991-1994, 48 RS cases were reported to the CDC and 93 RS hospitalizations based on HCIA data. When the HCIA data are extrapolated to the US population, there were an estimated 284 hospitalizations. Cases reported from both data sources were similar in distribution by onset, age, and sex. CDC data probably underestimate the incidence of RS due to incomplete reporting and HCIA data may overestimate it because not all cases were known to meet the CDC case definition. The true annual incidence of RS during the study years was probably between 0.2 and 1.1 cases per million population <18 years of age. PMID- 11060510 TI - Life crisis and the body within. AB - The authors take into account body involvement during the existential critical periods. The concept of body involvement considers both the biological (organic) involvement of the body, i.e. all the 'psychosomatic' (in a wider sense) dimensions, and the involvement of the mental representation of one's body. This involvement refers to some clinical conditions as 'abnormal somatic styles of existence', hypochondriasis and dysmorphophobia (all grouping in the DSM-IV 'somatoform disorders') and the anorexic states. From a structural psychogenetic point of view, this firm involvement of the body (naturally we are referring to the 'experienced body') is stressed in light of its importance in self identity construction and maintenance, especially when, as in some existential critical periods, identity physiologically faces significant modifications and hazardous movements. PMID- 11060511 TI - Psychological disorders in survivors of torture: exhaustion, impairment and depression. AB - Post-traumatic stress disorder (PTSD) has been described as the characteristic sequel to extreme events in life such as war and especially torture. This limitation to a single approach in regard to diagnosis and treatment has been criticised as being a too narrow concept to describe the effects following extreme events in life, especially as most studies so far were limited to PTSD and a small range of symptoms or disorders. The study presents data on psychiatric disorders in a group of exiled survivors of torture presenting to an out-patient department for psychiatry. A DSM-III-R-based psychiatric interview, including the general assessment of functioning scale (GAF), an open list of symptoms and the Vienna diagnostic criteria in regard to depression were used to evaluate a broader range of possible sequels. The most frequent present diagnosis in 44 patients seen over a period of 3 years was PTSD (n = 40), but criteria for a present diagnosis of other disorders were fulfilled in 34 patients, even years after torture, mainly major depression or dysthymia (n = 26). Criteria for functional psychosis were fulfilled in 4 patients. Many patients reported symptoms not assessed by DSM-III-R criteria, including feelings of shame and guilt, and ruminations on existential fears. The impairment as indicated by the GAF (mean 59.1) correlated best with the presence of the endogenomorphic depressive axial syndrome, but not with duration of imprisonment, age or other factors. Research on sequels to extreme trauma should not be restricted to a simple diagnosis of PTSD, but should continue to look for a broader conceptualisation, including neglected categories like the axial syndrome, as PTSD is common, but might not be the only factor of importance for research and treatment. ICD-10 might offer a more adequate interpretation of sequels. PMID- 11060512 TI - Quality of life in depression and anxiety disorders: an exploratory follow-up study after intensive inpatient cognitive behaviour therapy. AB - Thirty-seven patients with depression and anxiety disorder, who participated in an intensive inpatient cognitive behaviour therapy program for 6 weeks, were interviewed before treatment and 6 weeks after the end of treatment; in addition to other measures, quality of life was assessed with the Berlin Quality of Life Profile. Substantial reduction in subjective quality of life, objective functioning and environmental assets was found at baseline. At follow-up, according to clinical global impression, 13.5% of the patients were very much improved, 45.9% much improved; in 26.3% only slight improvement and in 16.2% no improvement was reported. Quality of life changed for the better in areas like work and education, leisure, housing, social relations, psychological well-being and a global rating of satisfaction with life, but not in marital relations, health in general and in finances. PMID- 11060513 TI - Seasonal variation in suicide in a predominantly Caucasian tropical/subtropical region of Australia. AB - Seasonal variations in suicide were examined in a Caucasian population living relatively close to the equator. A spring/early summer peak, but no secondary autumn peak, was found for males. An autumn trough was found for females. No significant seasonal variation was found for rurality, distance from the equator, employment status, or methods of suicide. Post-mortem blood alcohol levels were higher in spring and summer, possibly reflecting socialization patterns. The modest associations are consistent with suggestion that climatic influences may produce greater variation in suicide rates where the climatic variation itself is greater. PMID- 11060514 TI - Pain complaints in depressed inpatients. AB - Pain complaints were assessed in 150 depressed inpatients at admission (D0), after 10 days (D10) and 28 days of treatment (D28) using the Symptom Check List, 90 items, revised version (SCL-90R). Pain complaints were present in 92% of patients at D0, several pain complaints being reported by 76% of patients. Headache and chest pain were more frequent in women, whereas myalgia and numbness were more frequent in men. Pain complaints were related to depressive and anxious complaints as assessed by the SCL-90R, but not to age, suicide attempts and depression severity as assessed by the psychiatrist. Pain complaints decreased between D0 and D10, whereas depression scores decreased both between D0 and D10 and between D10 and D28. As compared to responders to treatment at D28, nonresponders had lower Montgomery and Asberg Depression Rating Scale scores at D0 and higher pain complaint scores at D0 and D10. PMID- 11060515 TI - Clinical features of delusional beliefs in schizophrenic and unipolar mood disorders: a comparative study. AB - We assessed comparatively 13 clinical features of delusions in a sample of 132 deluded inpatients of both sexes with schizophrenic (n = 89) or unipolar mood disorders (n = 43). Patients with schizophrenic disorders exhibited higher levels of severity than those with unipolar depression with respect to the features of vagueness-illogicality, bizarreness, systematization, conviction, duration and affective incongruence, whereas the reverse held true with respect to the feature of emotional impact. Furthermore, the two diagnostic groups were compared to each other with respect to patients' scores on five dimensions of their delusions obtained through factor analysis, namely emotional and behavioral impact, cognitive disintegration, delusional certainty, volitional dyscontrol and affective inappropriateness. Schizophrenic patients exhibited higher levels of severity than depressives on the second and fifth dimensions, whereas the reverse held true with respect to the fourth one. Our results suggest that particular features of delusions as well as broader dimensions thereof, may assist in the differential diagnosis of unipolar depression with psychotic features from schizophrenic disorders. PMID- 11060516 TI - Depressive symptomatology and short-term stability at a Nigerian psychiatric care facility. AB - The aims of the study were to examine: the frequency of the subtypes and symptoms of depression, the stability of symptoms in 3 months and to compare the data with previous studies. Ninety-six consecutive patients (31 males, 65 females) fulfilling ICD-10 criteria were assessed. Compared with the WHO study, the core symptoms of depression were similar; pathological guilt and suicidal behaviour were less prevalent, and psychoticism was more prevalent. Overall, 61.5% had severe depression, 64.6% had somatic syndrome, and subjects were in remission at follow-up. The results were similar to those of a report from the hospital 26 years earlier. The purported rarity of guilt, self-depreciation, suicide and psychotic symptoms should not imply rarity of severe depression among Africans; for these are not its commonest symptoms. PMID- 11060517 TI - Relationship between diagnostic subtypes of depression and occupation in Japan. AB - The relationship between occupational groups and the subtypes of endogenous depression was investigated. 98 patients who were diagnosed as having endogenous depression and hospitalized in the psychiatric department of a medical school hospital over the 15-year period between 1981 and 1996 were divided into two occupational groups: 67 patients were classified as belonging to the industrialized society occupation group (e.g., office workers, managers, teachers and technicians) and 31 patients were classified as belonging to the traditional society occupation group (e.g., farmers and skilled manual workers). The unipolar to-bipolar depression ratio for the industrialized society occupation group was revealed to be higher than that of the traditional society occupation group. This finding seems to support the view that the incidence of unipolar depression has increased with the process of industrialization. However, bipolar depression is less likely to be affected by societal and cultural changes due to the probably biologically dominant etiology of this form of depression. PMID- 11060518 TI - Treatment decisions for the extremely premature infant. PMID- 11060519 TI - Additional confirmation of the lack of effect of intravenous immunoglobulin in the prevention of neonatal infection. PMID- 11060520 TI - Initial management of immune thrombocytopenic purpura in children: is supportive counseling without therapeutic intervention sufficient? PMID- 11060521 TI - Genes, brains, and development: linking preschool developmental disorders to medical science and care. PMID- 11060522 TI - Neurodevelopmental outcome after the fontan procedure in children with the hypoplastic left heart syndrome and other forms of single ventricle pathology: challenges unresolved. PMID- 11060523 TI - The vitamin A paradox. PMID- 11060524 TI - Treatment choices for extremely preterm infants: an international perspective. AB - OBJECTIVE: To compare treatment choices of neonatal physicians and nurses in 11 European countries for a hypothetical case of extreme prematurity (24 weeks' gestational age, birth weight of 560 g, Apgar score of 1 at 1 minute). STUDY DESIGN: An anonymous, self-administered questionnaire was completed by 1401 physicians (response rate, 89%) and 3425 nurses (response rate, 86%) from a large, representative sample of 143 European neonatal intensive care units. Italy, Spain, France, Germany, the Netherlands, Luxembourg, Great Britain, Sweden, Hungary, Estonia, and Lithuania participated. RESULTS: Most physicians in every country but the Netherlands would resuscitate this baby and start intensive care. On subsequent deterioration of clinical conditions caused by a severe intraventricular hemorrhage, attitudes diverge: most neonatologists in Germany, Italy, Estonia, and Hungary would favor continuation of intensive care, whereas in the other countries some form of limitation of treatment would be the preferred choice. Parental wishes appear to play a role especially in Great Britain and the Netherlands. Nurses are more prone than doctors to withhold resuscitation in the delivery room and to ask parental opinion regarding subsequent treatment choices. CONCLUSION: An extremely premature infant is regarded as viable by most physicians, whereas after deterioration of the clinical conditions decision-making patterns vary according to country. These findings have implications for the ethical debate surrounding treatment of infants of borderline viability and for the interpretation and comparison of international statistics. PMID- 11060525 TI - Survival and neonatal morbidity at the limits of viability in the mid 1990s: 22 to 25 weeks. AB - OBJECTIVE: We determined neonatal survival and morbidity rates based on both fetal (stillborn) and neonatal deaths for infants delivered at 22 to 25 weeks' gestation. STUDY DESIGN: Two hundred seventy-eight deliveries at 22 to 25 weeks' completed gestation were analyzed by gestational age groups between January 1993 and December 1997. Logistic regression models were used to identify maternal and neonatal factors associated with survival. RESULTS: The rate of fetal death was 24%; 76% of infants were born alive and 46% survived to discharge. Survival rates including fetal death at 22, 23, 24, and 25 weeks were 1.8%, 34%, 49%, and 76%; and survival rates excluding fetal death were 4.6%, 46%, 59%, and 82%, respectively. Logistic regression analyses showed that higher gestational age (P<.0002), higher birth weight (P<.001), female sex (P<.005), and surfactant (P<.003) were associated with neonatal survival. Cesarean section was associated with decreased survival (P <.006). CONCLUSION: Hospital neonatal survival rates of infants at the limits of viability are significantly lower with the inclusion of fetal deaths. This information should be considered when providing prognostic advice to families when mothers are in labor at 22 to 25 weeks' gestation. PMID- 11060526 TI - Preterm infants with low immunoglobulin G levels have increased risk of neonatal sepsis but do not benefit from prophylactic immunoglobulin G. AB - OBJECTIVE: In a prospective, randomized, placebo-controlled, multicenter study, we evaluated the prevention of neonatal infections with intravenous immunoglobulin G (IVIgG) prophylaxis for preterm infants (gestational age <33 weeks) with umbilical cord blood IgG levels < or =4 g/L. STUDY DESIGN: Intravenous IgG or placebo (albumin), 1 g/kg body weight, was given on days 0, 3, 7, 14, and 21 to 81 infants with umbilical cord blood IgG levels < or =4 g/L: (1) IVIgG group, n = 40, mean (SD) gestational age 27.5 (2.2) weeks and birth weight 1.06 (0.39) kg; (2) placebo group, n = 41, mean (SD) gestational age 27.7 (2.5) weeks and birth weight 1.13 (0.38) kg. Infants with umbilical cord blood IgG levels >4 g/L (n = 238) served as a separate comparison group. Neonatal infections according to European Society of Pediatric Infectious Disease criteria were monitored until 28 days of life. RESULTS: Infants with IgG levels < or =4 g/L at birth who received IVIgG had no significant reduction in infectious episodes or mortality rate when compared with those given placebo. However, infants with a serum concentration of IgG >4 g/L at birth had significantly fewer infectious episodes (culture-proven sepsis) than infants with low serum concentrations of IgG (< or =4 g/L) when compared at the same gestational ages (26 to 29 weeks, P <.003). CONCLUSIONS: Prophylactic immunotherapy with IVIgG did not improve the immune competence in preterm infants with low serum IgG concentrations at birth. We speculate that a spontaneously high serum IgG concentration at birth reflects placenta function and is an indicator of a more mature immune system capable of protecting the preterm infant against severe neonatal infections. PMID- 11060527 TI - The clinical course of immune thrombocytopenic purpura in children who did not receive intravenous immunoglobulins or sustained prednisone treatment. AB - OBJECTIVE: To demonstrate the result of watchful waiting without specific therapy in unselected children with acute immune thrombocytopenic purpura (ITP). STUDY DESIGN: Between May 1992 and October 1999, 55 consecutive children (aged 2 months to 16 years; 28 boys and 27 girls) with acute ITP did not receive intravenously administered immune globulin G (IVIG) or sustained prednisone treatment. Patients with extensive mucosal bleeding were given prednisone, 2 mg/kg/d, for 3 days. RESULTS: In 37 of 55 patients the initial platelet count was <10,000/microL. Ten of these patients had active mucosal bleeding. Five additional patients with bleeding had platelet counts between 10,000 and 20,000/microL. Four patients were given a 3-day course of prednisone. Chronic ITP occurred in 7 (13%) of the patients; 29 patients achieved remission within 6 weeks, and 19 patients, between 6 weeks and 6 months. No life-threatening bleeding occurred, and no patient died. CONCLUSION: Most children with severe thrombocytopenia do not have active mucosal bleeding. This management approach, which did not administer specific therapy, avoided side effects, reduced cost, and was effective. PMID- 11060528 TI - Etiologic yield of single domain developmental delay: a prospective study. AB - OBJECTIVE: To determine the etiologic yield in young children with single domain developmental delay (either developmental language disorder or isolated motor delay) after a specialty diagnostic evaluation. METHODS: During an 18-month period, all children <5 years of age, who were consecutively referred to pediatric neurology or developmental pediatric clinics at a single tertiary pediatric center, were prospectively enrolled. Etiologic yield was determined after completion of clinical assessments and selected laboratory studies requested by the evaluating physician. RESULTS: Seventy-two children (60 boys) were found to have a developmental language disorder, and 22 children (11 boys) had isolated motor delay, of whom 6 had an associated diagnosis of cerebral palsy. An etiologic diagnosis was rarely made in the children with developmental language disorder (3/72, 4.1%). Laboratory investigations (metabolic, cytogenetic, imaging), aside from audiometry, were uniformly uninformative. In those children with isolated motor delay, an etiology was apparent in more than half (13/22, 59%). Slightly more than half (7/13, 54%) of etiologies identified in this group were potentially preventable. Successful etiologic determination in children with motor delay often had an impact on recurrence risk estimation, medical management, or specific therapy offered (8/13, 62%). The presence of physical findings on initial assessment was found to be highly predictive of successful etiologic determination in children with isolated motor delay (13/17 vs 0/5, P =.002). CONCLUSION: Etiologic yield differs substantially according to the subgroup of single domain developmental delay. PMID- 11060529 TI - Neurodevelopmental status of newborns and infants with congenital heart defects before and after open heart surgery. AB - BACKGROUND: Neurodevelopmental disabilities in children with congenital heart defects (CHDs) have been primarily attributed to intraoperative events without consideration of preoperative and postoperative factors. OBJECTIVE: To describe the preoperative and postoperative neurodevelopmental status of newborns and infants with CHDs. STUDY DESIGN: One hundred thirty-one children (56 newborns and 75 infants) were evaluated before and after surgery by using standardized neurobehavioral (newborn) and motor assessments (infant) and neurologic examinations. RESULTS: In newborns, neurobehavioral abnormalities were documented in >50% before surgery, with abnormalities persisting in most after surgery. In infants, neurodevelopmental abnormalities were observed in 38% before surgery. There was a significant association between preoperative and postoperative neurodevelopmental status, with status remaining unchanged in most. Newborns with acyanotic heart lesions were more likely to demonstrate neurologic compromise than those with cyanotic defects. For infants, arterial oxygen saturations <85% were significantly associated with an abnormality. There was a trend for a longer circulatory arrest time to be associated with greater risk for neurologic sequelae in newborns, whereas prolonged cardiopulmonary bypass was an important risk factor for infants. CONCLUSIONS: Neurodevelopmental abnormalities are common in young infants with CHDs and are often present before open heart surgery. These developmental concerns are clinically underappreciated. Early systematic developmental screening may be warranted in this population of interest. PMID- 11060530 TI - Neurodevelopmental outcome of patients after the fontan operation: A comparison between children with hypoplastic left heart syndrome and other functional single ventricle lesions. AB - OBJECTIVE: To compare neurodevelopmental outcome (NDO) in patients with hypoplastic left heart syndrome (HLHS), other functional single ventricle lesions, and the standard population and to investigate predictors of NDO in the population of children with functional single ventricle (FSV). STUDY DESIGN: A time- and age-defined cohort of patients with the Fontan circulation was recruited to participate in neurodevelopmental testing, behavioral evaluation, and imaging of the central nervous system. The Wechsler Intelligence test was the primary measure of NDO. Analysis included comparison of patients with HLHS with other patients with functional single ventricles. Other potential clinical predictors of NDO were investigated. RESULTS: The mean Full Scale Wechsler Intelligence score was 101.4+/-5.4. For the HLHS subgroup the mean Full Scale Wechsler score was 93.8+/-7.3, and for the non-HLHS subgroup it was 107.0+/-7.0. Although the HLHS group had significantly lower scores than the non-HLHS subgroup, neither subgroup scored significantly different from the standard population on the Wechsler Scales. Socioeconomic status, circulatory arrest, and perioperative seizures also were predictive of neurodevelopmental outcome. CONCLUSION: Neurodevelopmental and behavioral outcome in patients who have undergone the Fontan procedure including patients with HLHS is good in the preschool and early school years, with Wechsler Intelligence scores generally in the normal range. PMID- 11060531 TI - Outcome of preterm infants with congenital heart disease. AB - OBJECTIVES AND STUDY DESIGN: To evaluate the morbidity and mortality of preterm infants with congenital heart disease (CHD), a chart review was performed for infants with CHD, excluding isolated patent ductus arteriosus, who were <37 weeks' gestation, weighed <2500 g, and were admitted to our neonatal intensive care unit from 1976 to 1999 (N = 201). RESULTS: Patients in the study represented 1.9% of the total neonatal intensive care unit population <37 weeks' gestation and <2500 g. The median gestational age was 33 weeks, and the mean birth weight was 1852 g. CHD diagnosis frequencies were similar to those reported in other large incidence studies, except for a higher percentage of conotruncal defects. The risk of necrotizing enterocolitis was 1.7 times higher and the overall mortality twice as high in our patients compared with patients in the neonatal intensive care unit who did not have CHD. Cardiac surgery (n = 133) was performed on 108 patients. During the recent period of 1985 to 1999, compared with our institution's overall results for CHD surgery, the operative mortality rate was 10.4% versus 5.4% for closed procedures and 25.4% versus 10.5% for open procedures. The actuarial survival rate is 51% at 10 years; survival improved as the study period progressed. CONCLUSIONS: Infants with both CHD and prematurity did significantly worse than either group alone. Such outcome data are required for proper allocation of resources to care for this high-risk pediatric population. PMID- 11060532 TI - Vitamin A supplements and diarrheal and respiratory tract infections among children in Dar es Salaam, Tanzania. AB - OBJECTIVE: To determine the effect of vitamin A supplementation on the risk of diarrhea and of acute respiratory infection. DESIGN: Double-blind, randomized, placebo-controlled trial. SETTING: Dar-es-Salaam, Tanzania. SUBJECTS: Six hundred eighty-seven children, 6 to 60 months old, hospitalized with pneumonia, who received vitamin A or placebo at baseline and at 4 and 8 months after discharge from hospital. MAIN OUTCOME VARIABLES: Incidence and duration of episodes of diarrhea and respiratory tract infections during the year after discharge from the hospital. RESULTS: Relative to those receiving placebo, children receiving vitamin A had a significantly smaller risk of severe watery diarrhea (multivariate odds ratio = 0.56, 95% CI = 0.32-0.99, P =.04) but a higher risk of cough and rapid respiratory rate (multivariate odds ratio = 1.67, 95% CI = 1.17 2.36, P =.004). Vitamin A supplementation was also associated with increased risk of acute diarrhea among normally nourished children or children with stunted growth but was relatively protective among children with wasting disease (P value for interaction =.01). The apparently increased risk of respiratory tract infection was limited to children who were seronegative for human immunodeficiency virus (HIV) (P value for interaction =.07). CONCLUSIONS: Vitamin A supplements provide a low-cost intervention against morbidity in HIV-infected and undernourished children. Supplements may also have serious non-lethal adverse outcomes in well-nourished individuals. Whether these apparent detrimental effects of vitamin A are transient or long-term needs to be examined. PMID- 11060533 TI - Neonatal factors predicting childhood height in preterm infants: evidence for a persisting effect of early metabolic bone disease? AB - OBJECTIVES: Preterm infants are known to remain small during childhood. We previously found that evidence of neonatal metabolic bone disease was associated with reduced length at 18 months. We aimed to further investigate factors predicting childhood height and to test the hypothesis that evidence of early metabolic bone disease is associated with reduced later height. STUDY DESIGN: A cohort of preterm infants was measured prospectively at 18 months (n = 765), 7. 5 to 8 years (n = 772), and 9 to 12 years of age (n = 503). RESULTS: Preterm infants remained short for their age and sex at all follow-ups. Later height was most strongly predicted by parental height and, at 9 to 12 years, by pubertal status. Neonatal factors associated with later height were birth weight SD score, maternal hypertension/toxemia, and a high peak plasma alkaline phosphatase during the neonatal period. After adjustment for these factors plus interim heights, height at 9 to 12 years was greatest in those who were tallest at 7.5 to 8 years, those who had shown the greatest increase in height percentile between 18 months and 7.5 to 8 years and, as expected, those who were pubertal, whereas children with a peak neonatal plasma alkaline phosphatase >1200 IU were significantly shorter. CONCLUSIONS: Childhood height in preterm infants is strongly influenced by genetic factors. However, biochemical evidence of metabolic bone disease during the neonatal period may have a long-term stunting effect persisting up to 12 years later, providing support for current practices that aim to prevent this condition. PMID- 11060534 TI - Cutaneous manifestations of neonatal lupus without heart block: characteristics of mothers and children enrolled in a national registry. AB - OBJECTIVE: To extend the information base on cutaneous manifestations of neonatal lupus erythematosus (NLE) with regard to maternal disease, sex of child, onset, localization, influence of UV light, prognosis, and recurrence rates in subsequent pregnancies. METHODS: Review of records from the Research Registry for Neonatal Lupus. RESULTS: The cohort includes 47 mothers (83% white) whose sera contain anti-SSA/Ro, anti-SSB/La, and/or anti-U1-ribonucleoprotein antibodies and their 57 infants (20 boys and 37 girls) diagnosed with cutaneous NLE (absent heart disease) between 1981 and 1997. At detection of the child's rash, 13 mothers were asymptomatic, 11 had an undifferentiated autoimmune syndrome (UAS), 9 had systemic lupus erythematosus (SLE), 7 Sjogren's syndrome (SS), 6 SLE/SS, and 1 rheumatoid arthritis/SS; 20 reported photosensitivity. Within 5 years, 7 asymptomatic mothers experienced disease progression: 1 developed photosensitivity, 2 SLE, 3 SS, 1 SLE/SS; in 2 mothers UAS progressed to SLE; and 2 mothers with SS developed SLE. The infant's rash often followed UV light exposure; mean age at detection was 6 weeks, and mean duration was 17 weeks. All had facial involvement (periorbital region most common) followed by the scalp, trunk, extremities, neck, and intertriginous areas. In 37, the rash resolved without sequelae, 43% of which were untreated. A quarter had residual sequelae that included telangiectasia and dyspigmentation. One child developed Hashimoto's thyroiditis, and 2 developed systemic-onset juvenile rheumatoid arthritis. Of 20 subsequent births, 7 children were healthy, 2 had congenital heart block (CHB) only, 4 CHB and skin rash, and 7 skin rash only. CONCLUSIONS: Future pregnancies should be monitored by serial echocardiograms, given the substantial risk for heart block. Affected children should be observed for later development of a rheumatic disease. PMID- 11060535 TI - Age at symptom onset predicts severity of motor impairment and clinical outcome of glutaric acidemia type 1. AB - OBJECTIVES: In patients with glutaric acidemia type 1 (GAI), biochemical and molecular markers fail to predict the course of individual patients; therefore we sought to identify nonbiochemical variables that correlate with severity of motor deficits or overall clinical outcome. STUDY DESIGN: Archival data was collected from 42 published articles describing 115 patients with GA1. A forward, stepwise, multiple regression analysis was used to find predictors for outcome. RESULTS: Analyses show that in patients who did not have a precipitating illness before the first appearance of motor symptoms, the age at onset was significantly associated with the severity of motor impairments and overall clinical outcome. In patients who had a precipitating illness, the age at onset did not predict the outcome. In both groups of patients, basal ganglia degeneration, enlargement of spaces containing cerebrospinal fluid, and white matter abnormalities were indicative of a poorer prognosis. Treatment given after the appearance of symptoms was not associated with a better clinical outcome or fewer motor deficits. CONCLUSION: Because the age at symptom onset can significantly predict the severity of motor deficits and the overall outcome, it is important to identify patients with GA1 as early as possible. Several studies suggest that presymptomatic treatment may prevent or postpone the onset of symptoms. PMID- 11060536 TI - Long-term follow-up and outcome of 39 patients with chronic granulomatous disease. AB - OBJECTIVES: To evaluate the clinical long-term course in patients with chronic granulomatous disease (CGD) with respect to different CGD subtypes and currently used antimicrobial prophylactic measures. STUDY DESIGN: The records of 39 patients with CGD who were monitored during a period of 22 years were reviewed. All infections, infectious complications, and clinical outcomes were documented for a total observation period of 610 patient-years and were stratified with respect to different CGD subtypes. RESULTS: Lymphadenitis, skin abscesses, and pneumonia occurred in 87%, 72%, and 59% of the patients, respectively. In 151 microbiologic isolates Staphylococcus aureus, Aspergillus species, Candida species, Pseudomonas species, and Salmonella species were the most frequently detected microorganisms. There were 167 severe infections requiring hospitalization and intravenous antimicrobial treatment, resulting in an incidence of 3.7 severe infections per 100 patient months (SI/100 PM). Long-term antibiotic prophylaxis significantly reduced the incidence of severe bacterial infections from 4.8 SI/100 PM to 1. 6 SI/100 PM (P =.0035). In contrast, fungal infections increased under antibiotic prophylaxis from a mean incidence of 0.2 SI/100 PM to 1.9 SI/100 PM (P =.04). We found a 50% survival rate through the fourth decade of life, with a plateau after the third decade of life. Patients with a complete absence of cytochrome b(558) showed an earlier manifestation of their disease and a higher incidence of infections and had significant lower survival than patients with only diminished cytochrome b(558) or autosomal recessive CGD. CONCLUSIONS: Infections with Aspergillus species have become the major cause of infectious complications and death in patients with CGD. Prophylactic and therapeutic measures are needed to further increase life expectancy and quality for patients with CGD. PMID- 11060537 TI - Knowledge of Centers for Disease Control and Prevention guidelines for the use of vancomycin at a large tertiary care children's hospital. AB - OBJECTIVE: In 1994, the Centers for Disease Control and Prevention (CDC) published guidelines to encourage prudent use of vancomycin. We sought to determine whether physicians could demonstrate knowledge consistent with the guidelines. DESIGN: Survey consisting of 18 clinical vignettes based on the CDC guidelines. PARTICIPANTS: All residents, fellows, and attending physicians involved in pediatric inpatient services. SETTING: Tertiary care children's hospital providing service to an inner-city population and community referral base. MAIN OUTCOME MEASURES: Comparison of survey scores and individual responses among respondents. RESULTS: Survey scores did not vary with level of training or whether the respondent was a pediatrician or non-pediatrician. Average scores of attending physicians, fellows, and residents were 74.1% (SD = 13.1), 77.2% (SD = 11.5), and 73.4% (SD = 10.5), respectively, and did not differ significantly. Questions incorrectly answered by more than 30% of respondents concerned the use of vancomycin as: (1) first-line treatment of Clostridium difficile colitis, (2) a topical solution for wound infection, (3) initial, empiric treatment of patients with fever and neutropenia, (4) peri-operative prophylaxis, (5) a preferred agent over beta-lactam antimicrobial agents. CONCLUSION: Deficits in knowledge regarding appropriate vancomycin use can be localized to certain clinical settings. This observation lends optimism to the notion that targeted educational intervention may improve the appropriate use of vancomycin. PMID- 11060538 TI - Impact of extreme prematurity on families of adolescent children. AB - OBJECTIVES: To compare the impact of burden of illness on families of teenaged children who were extremely low birth weight (ELBW) with that of members of a term control group (C) and to determine whether the attitudes toward active treatment of very immature infants differ between the 2 cohorts. DESIGN: In a cross-sectional survey, parents of 145 (86%) of 169 members of an ELBW cohort and 123 (85%) of 145 members of a control cohort completed a 23-item self-completed questionnaire encompassing occupational, marital, and family-related issues and attitudes toward treatment of infants of borderline viability. RESULTS: Both positive (P =.0003) and negative (P <.005) effects on marriage were higher in parents of the ELBW group; although more parents in the ELBW group felt that their child had brought their families closer together (P =.0001), their child's health had adversely affected their emotional health (P =.02) and that of other children in the family (P =.003). Despite this result, a significant proportion of parents from both cohorts supported saving all infants (ELBW 68%; C 58%) and favored the role of parents in decision making (ELBW 98%; C 97%). CONCLUSIONS: In the long term, it appears that parents of ELBW children have adjusted fairly well to their work and family life. Although some negative effects were identified, there was still considerable support for active treatment of infants of borderline viability. PMID- 11060539 TI - Exposure to violence and victimization, depression, substance use, and the use of violence by young adolescents. AB - OBJECTIVE: To examine the relationships among exposure to violence; tobacco, alcohol, and other substance use; depression; church attendance; and the use of violence among very young adolescents. METHODS: An 86-item confidential questionnaire was administered to 722 sixth grade students (mean age = 11.9+/-0.8 years) attending 4 middle schools serving neighborhoods in and around public housing. RESULTS: Boys had a higher mean violence scale score than girls (P < or =.0001), and students living in public housing had higher violence scale scores than other students (P< or =.0001). Self-reported use of violence was significantly associated with exposure to violence (r =.45); age (r =.28); frequency of church attendance (r = -.14); depression (r =.28); the probability of being alive at age 25 (r = -.09); the frequency of use of cigarettes (r =.39), alcohol (r =.37), and multiple substances (r =.38); and interest in a gang (r =.37). When all of these variables were analyzed with multiple linear regression, multiple substance use, exposure to violence, interest in a gang, male gender, cigarette smoking, and depression level accounted for 49.7% of the variation in the use of violence scale. CONCLUSION: Recent multiple substance use and lifetime exposure to violence and victimization were the strongest correlates with the frequency that these youth reported using violence and carrying weapons. PMID- 11060540 TI - Protein C deficiency related to valproic acid therapy: a possible association with childhood stroke. AB - We report a case of stroke in a child with acquired protein C deficiency receiving valproic acid (VPA). To investigate the possible association of VPA with protein C deficiency, protein C levels were measured in 20 children receiving VPA monotherapy and 20 children receiving other anticonvulsants. Protein C levels were reduced in up to 45% of the VPA-treated subjects. PMID- 11060541 TI - Severe hepatic Wilson's disease in preschool-aged children. AB - A 3-year-old girl presented with hemolytic anemia, hepatosplenomegaly, ascites, and evidence of decompensated chronic liver disease. Genotypic DNA analysis revealed that the patient was homozygous for a splice site mutation now designated IVS4-1:G>C, expected to destroy completely the functional gene product of ATP7B, the gene responsible for Wilson's disease. We suggest that this severe mutation caused very early liver disease. Wilson's disease should be considered in the differential diagnosis of established liver disease in the preschool-aged child. PMID- 11060542 TI - Treatment of severe complicated Kawasaki disease with oral prednisolone and aspirin. AB - We report on 7 patients with severe, complicated Kawasaki disease treated with oral prednisolone, after apparently unsuccessful intravenous immunoglobulin treatment. An additional eighth patient was a Jehovah's Witness, who was given steroid and aspirin as first-line treatment. These findings support a beneficial role for steroids in intravenous immunoglobulin-resistant Kawasaki disease. PMID- 11060543 TI - Children with morphea have normal self-perception. AB - We evaluated the self-esteem and quality of life of 47 children with morphea with the use of the Harter self-perception profile for children and Visual Analog Scale. Most children with morphea have normal self-worth and a high quality of life. Morphea, like some other childhood chronic illnesses, does not impair self esteem. PMID- 11060544 TI - A decimillennium in neonatology. PMID- 11060545 TI - Reducing heat loss at birth in very preterm infants. PMID- 11060547 TI - Correction. Effect of polyethylene occlusive skin wrapping on heat loss in very low birth weight infants at delivery: a randomized trial. PMID- 11060549 TI - Evaluating and monitoring the adolescent with pulmonary function testing. AB - This article reviews pulmonary function tests (quantitative measurements of physiologic functioning of lungs, chest wall, and respiratory muscles) as valuable tools for the diagnosis and management of chronic respiratory disease in adolescents. Advances in technology have made pulmonary function testing more available and, for some tests, easier to perform. Pulmonary function evaluation is used for diagnosis of pulmonary disease. Testing will identify abnormal physiologic processes that result in obstructive or restrictive pulmonary disease and, together with a complete history, physical examination, and other tests, help to determine the specific etiology. Pulmonary function measurement may be used to manage a patientis disease over time. Quantitative evaluation of the response to many specific therapies, such as asthma medications and environmental control, can only be achieved by measuring pulmonary function. Monitoring pulmonary toxicity of medications, such as chemotherapeutic agents, is also done by measuring pulmonary function. In many patients with chronic pulmonary disease, such as cystic fibrosis, the pulmonary function laboratory can be used to assess the patient's physiologic limitation. PMID- 11060550 TI - Medical care of the adolescent with asthma. AB - Asthma is the most common chronic disease in adolescents. Despite advances in the understanding of this disease and the availability of more specific treatment, the prevalence of asthma and its morbidity and mortality are increasing. This trend is especially prominent and worrisome in the age group that includes adolescents and young adults. Possible factors contributing to this significant problem in adolescents include a lack of knowledge about the disease, delays in seeking medical attention due to denial of symptoms or overuse of acute relief medication that mask the progression of the inflammation, and various psychological problems such as low self-esteem or depression. These factors, in addition to the typical developmental behaviors recognized in adolescence, contribute to the complexity of asthma management in this age group. This article comprehensively reviews the pathophysiology and precipitating factors of asthma as well as all aspects of medical care of affected individuals, including monitoring and self-care. PMID- 11060551 TI - Allergy evaluation and immunotherapy. AB - Up to 10% of children and adolescents in the United States may have asthma and up to 40% may be affected by allergic rhinitis. Most people know that asthma is a serious disease, but "hay fever" is often mistakenly considered trivial. However, hay fever symptoms can significantly influence a patient's quality of life, causing fatigue, headache, and even cognitive impairment. Both asthma and allergic rhinitis can result in lost sleep, many missed school days, and the inability to participate in sports and other recreational activities in which young people engage. An appropriate diagnosis is the first step toward improving quality of life for these young patients. This article reviews diagnostic procedures for allergic disease, and-because compliance is frequently an issue with school-aged patients-it also reviews current thinking on allergen immunotherapy, a treatment that provides effective long-term control in appropriately selected patients. PMID- 11060552 TI - Use of complementary and alternative medicine in the treatment of asthma. AB - Complementary and alternative therapies are a growing part of our nation's health care and culture. The numbers of people using such forms of treatment are increasing, especially for patients with chronic diseases. There are an estimated 15 million prescription drug users nationwide who also use herbal medication. At the same time, asthma has increased in prevalence and severity, despite the increase in therapeutic options. Asthma patients and their parents may be interested in complementary medicine because of the long-term nature of their illness and perceived toxicities of therapies such as inhaled corticosteroids. Many alternative nutritional therapies are available in stores without a prescription, and this may also appeal to an adolescent's desire for independence. This article examines the phenomenon of alternative medicine, the possible reasons for its rising prevalence, and who may be using it. The emergence of increasing numbers of patients with asthma and the pathophysiology of this disease is discussed briefly. Finally, individual therapies with their proposed mechanism of action and known studies related to their use in asthma are examined. PMID- 11060553 TI - Respiratory diseases with a psychosomatic component in adolescents. AB - Adolescence is often a time of emotional upheaval and it is no wonder that many respiratory diseases with a psychosomatic component find their origins or time of exacerbation during this time of life. Adolescents who present with unexplained respiratory diseases may also suffer from some form of psychosomatic illness. Recognition of the psychological contributions to symptoms related to the respiratory tract is essential for practitioners who care for adolescents. This article includes some of the more common respiratory or related conditions that have psychological etiologies or components and are encountered in the adolescent patient. These include psychogenic cough, sighing dyspnea, hyperventilation, vocal cord dysfunction, and emotional state as a trigger for asthma. This review provides a general discussion of these conditions and an overview of issues related to psychological/psychiatric evaluation and the reluctance of patients and their parents to access mental health treatment. PMID- 11060554 TI - Pulmonary effects of smoking. AB - The problems of tobacco addiction have evolved over centuries. The possible relationship between smoking and oral cancer was recognized as early as the 19th century. The use of tobacco results in an estimated 4 million deaths each year worldwide. Approximately 3,000 adolescents start smoking every day; 4.5 million children and adolescents smoke cigarettes; 1 million use smokeless tobacco. This article reviews the effects of environmental tobacco smoke and primary smoking on lung health and maturation and the pathophysiology of smoking-related pulmonary disease. Smoking prevention and timely smoking cessation will significantly reduce the risk of not only lung diseases (chronic obstructive pulmonary disease, cancer, chronic bronchitis, asthma, etc.) but also suboptimal lung growth during preadolescent and adolescent years. PMID- 11060555 TI - Office interventions for adolescent smokers. AB - Smoking-prevention efforts can be undertaken at a national level, with enactment and enforcement of laws on the use of tobacco products by youth; at the state and local level, with involvement of community organizations; and through school systems, with education regarding the harmful effects of tobacco use. This review, however, focuses on the role of individual practitioners who also can make significant contributions by working at an individual level to incorporate prevention and treatment strategies in their daily medical practice. This article reviews two types of smoking cessation interventions-behavioral and pharmacologic. Currently available data on the prevention and treatment of nicotine addiction in adolescents, particularly pharmacotherapy, are quite limited. The individual clinician can contribute to prevention and treatment of tobacco use among children and adolescents by using many of the known behavioral and pharmacologic strategies. PMID- 11060556 TI - Cystic fibrosis in adolescents and young adults. AB - Advances in knowledge and medical science have resulted in an increased life span and quality of life of patients with cystic fibrosis (CF). The median age of survival for CF patients is 32.3 years of age and patients 18 years of age or older now constitute one third of the total patients with CF. Because of these advances, a new patient population has emerged: the adolescents and young adults with CF. Adolescence is normally a time of great cognitive, social and developmental changes. Adolescents with CF not only have to deal with the normal changes expected, but also have to deal with the transition of assuming responsibility for their care from the parents and transitioning their care from a pediatric to an adult care team. Moreover, many of these young adults have to deal with the impact of the progressive deterioration of their CF disease. This review discusses issues of significance to this emerging patient population, including medical care, issues of disability, and psychosocial and other medical conditions associated with an increased life expectancy. PMID- 11060557 TI - Evaluation and treatment of sleep disorders in adolescents. AB - Sleep disorders are quite common in adolescents and sleep disturbances occur most often during the mid-adolescent years. These disorders result in considerable negative consequences in terms of increased accidents, decreased academic performance, and increased behavioral difficulties. Over the past 25 years, the evaluation, diagnosis, and treatment of sleep-related breathing disorders has undergone dramatic changes. The diagnosis and treatment of pediatric sleep disorders are now the focus of more complex and collaborative research efforts. This article discusses the differences between childhood and adolescent sleep cycles, delayed sleep phase syndrome, sleep-related breathing disorders, narcolepsy, parasomnias, and behavior-related sleep problems. PMID- 11060558 TI - Respiratory problems in the adolescent with developmental delay. AB - Patients with developmental disorders, including adolescents, comprise a large and heterogeneous group of individuals who vary in underlying diagnosis and degree of disability. The largest numbers of patients are those with cerebral palsy and with traumatic brain injury. While these conditions themselves do not directly cause airway or parenchymal lung dysfunction, consequences of neuromuscular dysfunction, especially aspiration and ineffective cough, may lead to lung damage. Poor nutritional status, impairment of airway clearance by muscular weakness or incoordination and poor pulmonary reserve (due to chest wall or spine deformity) increase the risk of significant morbidity and mortality from respiratory infections. Individuals who were premature infants or who had prolonged neonatal courses may also have residual chronic lung disease (bronchopulmonary dysplasia) contributing to their pulmonary problems. This review discusses conditions that have adverse effects on the airway and lung (drooling, feeding problems, gastroesophageal reflux, aspiration, spasticity, scoliosis) and some of the consequences of these insults (disordered airway clearance, pneumonia, sleep apnea). Also discussed are issues important to the prevention or amelioration of respiratory difficulties, including preventive care, the effects of exercise, dental hygiene, and surgical intervention. PMID- 11060559 TI - Pulmonary complications in neuromuscular disease. AB - Pulmonary compromise can result from primarily peripheral or central neuropathic disease (including neurodegenerative diseases) or diseases of the muscle or neuromuscular junction. Some diseases present with a combination of nervous system and muscle tissue involvement (due to secondary changes like atrophy or primary pathology, as in mitochondrial diseases). While clinically different in pathophysiology, these diseases may progress to produce and isharei common pulmonary pathology by the time the patient reaches the adolescent years. This article presents a review of the categories of neurologic disease that may lead to respiratory compromise, followed by thorough description of the pulmonary complications that can result from these types of diseases. Finally, a review of currently accepted treatment options that may aid in improving the quality of life of these patients is offered. PMID- 11060560 TI - Gastroesophageal reflux in the adolescent. AB - Gastroesophageal reflux (GER) is relatively common in adolescence. The severity of gastrointestinal symptoms associated with gastroesophageal reflux varies from an occasional burp to persistent emesis. Evaluation of most of these patients reveals no definable anatomic, metabolic, infectious, or neurologic etiology. The clinical determination of a cause-and-effect relationship between GER and other disorders, including associated respiratory disease, is often difficult and must be approached with considerable caution. Tests that merely document the presence of GER add little to the diagnosis. The adolescent with GER often has persistent symptoms of esophagitis that lead to appropriate intervention. Understanding the capabilities and limitations of the various diagnostic maneuvers available to assess GER is important to avoid subjecting these patients to invasive, costly, and inappropriate testing. This article includes a general discussion of physiology, diagnostic evaluation, and therapy of GER, followed by a review of respiratory and other complications. PMID- 11060561 TI - Pulmonary infections in the adolescent with immunodeficiency. AB - Pulmonary infections in the immunocompromised adolescent are important causes of significant morbidity and mortality. Advances in the fields of chemotherapy, organ transplantation, and use of corticosteroid and immunosuppressive therapies have led to significant improvement in the outcome of patients with malignancy and a variety of other disorders. HIV infection has become a major additional cause of immune suppression. In turn, physicians are encountering growing numbers of patients with an impaired immune system presenting with respiratory and other infections. This article presents a brief review of defense mechanisms in the respiratory tract, selected conditions leading to impaired immune responses in the adolescent, specific pulmonary pathogens and their evaluation in the immunocompromised adolescent, and general considerations of the management of pulmonary infections in the immunocompromised adolescent. Infections of the upper respiratory tract and pulmonary infections seen in adolescents with cystic fibrosis are not discussed. PMID- 11060562 TI - Community-acquired pneumonia in adolescents. AB - Community acquired pneumonia (CAP) is defined as pneumonia acquired outside of the hospital setting. Extensive studies of CAP in adolescents that characterize the true incidence of various etiologic pathogens are not available. However, Streptococcus pneumoniae, Mycoplasma pneumoniae, and Chlamydia pneumoniae appear to be the most frequently encountered pathogens. These organisms often cause CAP in adults as well; other infections are noted as well, including Legionella. "Atypical pneumonia" refers to pneumonia not presenting with the usual clinical picture of pneumococcal infection (which includes high fever, productive cough, chills, and other "classic" features). The term is frequently used in adolescents with CAP. However, this classification may not help in individual patients, who often show a high degree of variability in the clinical presentation of pneumonia; also it does not always predict microbial cause. There is currently a trend away from the concept of atypical pneumonia syndrome and more discussion of atypical pathogens as commonly causes of CAP. This article reviews recent literature on CAP with special emphasis on its diagnosis and management in adolescent patients. PMID- 11060563 TI - Unexpected mortality reduction with abciximab for in-stent restenosis. AB - Treatment of in-stent restenosis is generally considered low risk, and it is not clear if adjunctive use of abciximab is beneficial in this low-risk population. We determined the effect of adjunctive abciximab during percutaneous coronary intervention (PCI) for treatment of in-stent restenosis. Two hundred and ninety three patients with in-stent restenosis underwent PCI at the Cleveland Clinic between January 1996 and December 1998. Patients undergoing directional atherectomy, laser treatment and brachytherapy were excluded (9 patients). Of the remaining 284, seventy-nine patients received abciximab during PCI and 205 were treated without abciximab. The groups were similar with respect to age, gender, left ventricular function, number of vessels involved, history of prior coronary artery bypass grafting and unstable symptoms at presentation. There were more diabetics, hypertensives, and patients with elevated cholesterol in the abciximab treated group. At 1-year follow-up, there was a significantly lower incidence of myocardial infarction (2.5% versus 5.3%; p < 0.05) and lower mortality (1.2% versus 5.8%; p < 0.01) in the abciximab-treated group. There was no difference in the incidence of revascularization. The findings of a lowered mortality and myocardial infarction rate with abciximab warrants further prospective study in patients with in-stent restenosis. PMID- 11060564 TI - Avoidance of adverse events in restenotic lesions with abciximab? PMID- 11060565 TI - Transradial coronary angioplasty and stent implantation in acute myocardial infarction: initial experience. AB - Transradial coronary angioplasty and stent implantation in have been associated with reduced complications, length of stay and hospital costs when compared to the transfemoral approach. Fourteen high-risk patients with acute myocardial infarction underwent transradial coronary angioplasty and stent placement. All diagnostic and interventional procedures were successfully completed using 6 French guide catheters and ACS Tristar stents (Guidant Corporation, Santa Clara, California) up to 4 mm in diameter. Thirteen patients received glycoprotein IIb/IIIa inhibitors. There were no procedural or access site complications. The mean length of stay was 3.5 days and the mean time interval from initial radial cannulation compared favorably with 14 acute myocardial infarction patients undergoing transfemoral angioplasty and stent placement. Transradial angioplasty in acute myocardial infarction appears to be a safe and feasible option. The procedure time is not increased in experienced hands, and the combination of rare access site complications and early ambulation may lead to decreased morbidity and lower costs. Transradial angioplasty in acute myocardial infarction may be an attractive option in thrombolytic therapy patients (facilitated percutaneous coronary intervention) or those who require aggressive anticoagulation or antiplatelet therapy. PMID- 11060566 TI - Transradial coronary angioplasty and stenting: an alternative to the femoral approach? PMID- 11060567 TI - Is radial stenting ready for the "primary" role? PMID- 11060568 TI - Effects of 308 nanometer excimer laser energy on 316 L stainless-steel stents: implications for laser atherectomy of in-stent restenosis. AB - PURPOSE: To determine the effects of the incidental exposure of stents to pulsed 308 nanometer ultraviolet excimer laser energy. METHODS: Five types of 316 L stainless-steel coronary stents were subjected to two types of study. First, for endurance testing, sixty stents were deployed in 3.0Eth 4.0 mm polymer tubes in three geometries. Up to 1,000 laser pulses were delivered while advancing a 2.0 mm eccentric catheter through the lumen of the stent. These stents were next subjected to 400 million simulated heartbeats and then analyzed for metal etching and fatigue. Second, six additional stents were irradiated with 1,000 pulses underwater and then analyzed for particulates, anions and cations liberated from the stent. RESULTS: Photomicroscopy revealed surface etching on a number of stents. Two stent models exhibited multiple strut fractures at the strut joints in both test samples and controls. In no case was a break observed at the site of laser-stent interaction. Breakage frequency was not significantly different between lazed stents and controls. Lazed stents produced a mean of 14 micrograms of sodium and 4 micrograms of iron more than controls. No excess particulates were detected. CONCLUSION: Under model conditions typical of clinical use, excimer laser treatment does not alter stainless-steel stent endurance or liberate clinically significant material from the stent. PMID- 11060569 TI - Efficacy and safety of direct stenting in coronary angioplasty. AB - BACKGROUND: Direct stenting is the deployment of an intracoronary stent without lesion predilation. Potential advantages include shorter procedural time, lower contrast dose and reduced spiral dissections. There is also the potential financial benefit of less balloon and/or stent usage. Concern still exists among some operators, however, regarding failure of stent deployment and local complications. METHODS: Of 467 consecutive angioplasty cases at the Alfred Hospital between August 1, 1997 and May 22, 1998, direct stenting was attempted in 93 patients (20%). Interventionalist preference determined whether direct stenting was attempted. Vessels with excessive calcification, severe proximal tortuosity or small caliber were typically considered unsuitable for direct stenting. RESULTS: A total of 102 lesions (38 type A, 60 type B, and 4 type C) were treated with direct stenting. Initial deployment was successful in 98 of 102 lesions, with a further 3 lesions successfully stented following predilation. A stent was unable to be deployed in only 1 case; however, the lesion was treated with balloon angioplasty alone. The majority of lesions required only 1 stent (an average of 1.1 stents were used per lesion). Distal complications occurred in 5 patients. In 3 patients, a small distal dissection was successfully stented, and in 1 case embolization of debris occurred down the distal vessel, resulting in a small procedural myocardial infarction. Only 1 patient out of 93 (1%) developed a large distal dissection requiring the deployment of multiple stents, compared with 22 of the remaining 374 patients (5.9%) who underwent conventional angioplasty. This was a significant difference in favor of direct stenting (Chi square, p < 0.05). When compared with a cohort of patients matched by lesion grade treated with conventional stenting, direct stenting used significantly less contrast per case (154 +/- 7.6 ml compared with 202 +/- 9.5 ml for conventional stenting; p = 0.0001). CONCLUSION: Direct stenting is a safe and effective method for treating coronary artery disease. In appropriately selected cases, it has a low rate of procedural failure and results in less contrast usage and fewer distal complications than conventional angioplasty and stenting. PMID- 11060570 TI - Accuracy of biplane analysis of left ventricular ejection fraction based on two consecutive single plane studies. AB - BACKGROUND AND PURPOSE: Mainly due to the high costs of biplane equipment many cardiac laboratories run single plane angiographic equipment only. Consequently, a biplane ventriculogram may only be done with two consecutive single plane studies. The aim of this investigation was to assess the accuracy of a biplane analysis of two consecutive single plane studies. METHODS: A total of 42 patients (62 +/- 10 years, 76% males), able to tolerate two consecutive ventriculograms without arrhythmia during the first study underwent two consecutive biplane studies (LAO 60, RA0 30), using 40 ml of contrast each. After the first injection, the x-ray tube was moved in a neutral position, and then was replaced in the 30 RAO/60 LAO position. Digital data was analyzed by two separate investigators using commercially available software. RESULTS: Intra-observer variability of left ventricular ejection fraction (LVEF) showed a high degree of agreement (single plane 1 vs. 2: r = 0.98; standard error of regression (Sy.x.): 2.8); the variability was slightly higher with two investigators (single plane: r = 0.92, Sy.x: 5.5 ) and with biplane analysis (biplane 1 vs. 2: r = 0.90, Sy. x: 5.7). End-diastolic volume index (EDVI) increased significantly from the first to the second study (84 +/- 28 ml/m2 vs 87 +/- 30 ml/m2; p = 0.017): Still LVEF of the two consecutive biplane studies showed very good agreement (biplane 1 vs. 2: mean difference (MD), -1.0; standard deviation of the difference (SDD), 5.2%). This agreement was almost as good as the one of LVEF values calculated from two consecutive single plane, but biplane analyzed studies compared to simultaneous biplane studies (MD, -0.5; SDD, 4.3%). CONCLUSION: Despite the significant increase in EDVI after contrast injection, LVEF values determined from two consecutive studies remained virtually unchanged. Biplane analysis of LVEF values based on consecutive single plane studies resulted in similar and reliable values as determined by two consecutive biplane studies. PMID- 11060571 TI - 'Cold hand, ischemic heart': treatment by stenting of the left subclavian artery. AB - A 59-year-old man presented with worsening angina and a cold, painful left hand, eight years after coronary artery bypass surgery. Coronary angiography showed extensive coronary atherosclerosis with blocked vein grafts to his left circumflex and right coronary arteries. There was a severe narrowing in the left subclavian artery before the origin of the left internal mammary artery (LIMA) which appeared patent. PTCA and stent implantation to the left subclavian artery stenosis restored normal flow to the left hand and the LIMA with abolition of his ischemic hand symptoms and marked improvement of his angina. PMID- 11060572 TI - Successful treatment of vasospastic angina with a coronary stent. AB - Calcium antagonists are the treatment of choice in vasospasm angina when no stenosis or mild stenosis are present. We present a case in which ergonovine echocardiography showed vasospasm of the right coronary artery despite optimal medical treatment. Stenting of a mild stenosis in that artery successfully controlled vasospasm and a pre-discharge ergonovine echocardiographic test was negative. The patient remains asymptomatic one year after stenting. PMID- 11060573 TI - Successful sealing of an angioplasty-related saphenous vein graft rupture with a PTFE-covered stent. AB - A patient developed severe hemodynamic compromise after saphenous vein graft rupture during high-pressure coronary stent deployment. Immediate balloon inflation followed by implantation of a polytetrafluoroethylene-covered stent solved the problem. The next day, follow-up angiography confirmed persistent sealing of the rupture site. Clinical and angiographic features of vessel rupture during coronary angioplasty as well as several therapeutic options for this complication are discussed. PMID- 11060574 TI - Choosing your allies. PMID- 11060575 TI - Canadian implantable defibrillator study: what does it mean after the antiarrhythmics versus implantable defibrillators trial? PMID- 11060576 TI - Prevention of ventricular arrhythmias in the congenital long QT syndrome. AB - Life-long therapy is necessary for patients with symptomatic long QT syndrome to prevent arrhythmic death. The merits and limitations of the different therapeutic modalities are discussed. beta-blockers remain the mainstay of therapy, but this medication may not be sufficient for cardiac arrest survivors and for those with the LQT3 genotype. "Genotype-specific" therapy, like potassium-channel openers for patients with inadequate potassium outflow (LQT1 and LQT2 genotypes) or sodium-channel blockers for patients with excessive sodium inflow (LQT3), significantly shortens the QT interval, but the effects of these drugs on arrhythmia prevention is less well established. Cardiac pacemakers may be especially beneficial for patients with LQT2 or LQT3 and for those with pause dependent torsade de pointes. More important is to recognize that device programming for preventing tachyarrhythmias in patients with long QT differs from the standard pacemaker programming. Finally, implantable defibrillators with dual chamber pacing capability are indicated for patients at high risk for arrhythmic death, including all cardiac arrest survivors. PMID- 11060577 TI - The Brugada syndrome. AB - The Brugada syndrome is a hereditary disease causing sudden cardiac death in apparently healthy individuals with a structurally normal heart. The disease is caused by mutations in the cardiac sodium channel gene SCN5A. Patients with this disease have a peculiar electrocardiogram with elevation of the ST segment in leads V1 to V3, an electrocardiogram that every doctor should recognize. There exist variants of the electrocardiogram with minimal ST segment elevation and even concealed forms that can only be unmasked by the administration of class I antiarrhythmic drugs. When left untreated or when treated with all known antiarrhythmic drugs, patients with Brugada syndrome have a high mortality (approximately 10% per year). The only effective treatment to prevent sudden death is the implantable defibrillator. PMID- 11060578 TI - Management of ventricular tachycardia in patients with clinically normal hearts. AB - The majority of patients who present with ventricular tachycardia have underlying structural heart disease. However, there has been increasing appreciation of the existence of multiple forms of idiopathic ventricular tachycardia with distinct features and unique mechanisms. The most common form of idiopathic ventricular tachycardia originates from the right ventricular outflow tract, is characterized by sensitivity to adenosine, and appears to be due to cyclic AMP-mediated triggered activity. Other forms of idiopathic ventricular tachycardia include intrafascicular left ventricular tachycardia, due to reentry, which is sensitive to verapamil, and automatic, propranolol-sensitive ventricular tachycardia. PMID- 11060579 TI - Ventricular arrhythmias, sudden death, and prevention in patients with hypertrophic cardiomyopathy. AB - Sudden unexpected death, often occurring in young asymptomatic patients, is the most devastating facet of the natural history of hypertrophic cardiomyopathy (HCM). It appears to be the consequence of primary ventricular tachyarrhythmias arising in an electrically unstable myocardial substrate characterized by disorganized cellular architecture, ischemia, cell death, and replacement scarring. Although identification of the HCM patient subset at high risk for a catastrophic event with precision continues to present challenges, treatment strategies for the prevention of sudden death are now available. In particular, the implantable cardioverter-defibrillator has a high degree of efficacy in sensing and terminating potentially lethal ventricular tachyarrhythmias and a life-saving role in both the primary and secondary prevention of sudden cardiac death in HCM. PMID- 11060580 TI - Catheter ablation of ventricular tachycardia related to coronary artery disease: the role of noncontact mapping. AB - There are a number of limitations associated with conventional mapping for ablation of ventricular tachycardia (VT) in ischemic heart disease, such as the high recurrence rates after initially successful ablation. The development of a noncontact mapping system capable of producing high-resolution isopotential maps of the entire left ventricle has enabled rapid identification of diastolic activity that maintains VT for ablation. With this system it is possible to map nonsustained and fast unstable as well as stable VTs. In this article we review the historic background and concepts of noncontact mapping, its clinical application, and the results of ablations for human VT guided by this mapping system. PMID- 11060581 TI - Strategies for catheter ablation of scar-related ventricular tachycardia. AB - Ventricular tachycardia (VT) due to reentry in and around regions of ventricular scar from an old myocardial infarction or cardiomyopathic process is often a difficult management problem. Radiofrequency catheter ablation is an option for controlling frequent VT episodes. Patient and VT characteristics determine the mapping and ablation approach and efficacy. In patients with a VT that is hemodynamically tolerated to allow mapping, prevention of recurrent VT is achieved in 54% to 66% of patients with a procedure related mortality of 1% to 2.7%. Multiple morphologies of monomorphic VT and circuits that are located deep to the endocardium are common problems that reduce efficacy. Mapping to identify target regions for ablation can be difficult if VT is rapid and not tolerated, or not inducible. Ablation of these "unmappable VTs" by designing ablation lines or areas based on the characteristics of the scar as assessed during sinus rhythm, and using approaches to assess global activation from a limited number of beats has been shown to be feasible. Ablation of multiple VTs, epicardial VTs, and poorly tolerated VTs are feasible. Future studies defining efficacy and risks are needed. PMID- 11060582 TI - Review of stroke in octogenarians undergoing coronary artery surgery with and without cardiopulmonary bypass. PMID- 11060583 TI - Minimal access aortic root, valve, and complex ascending aortic surgery. AB - We report our entire experience with minimal access aortic root, valve, and complex ascending aortic surgery. A total of 290 consecutive patients underwent aortic root, valve, and ascending aortic surgery between July 1996 and February 2000. Four groups were identified: isolated aortic valve replacement (AV group, n = 227), aortic root replacement (AR group, n = 44), aortic valve replacement with concomitant replacement of the supracoronary ascending aorta (V/A group, n = 9), and isolated ascending aortic replacement (AA group, n = 10). The procedures were performed through a partial upper hemisternotomy (87%) or a right parasternal approach (13%). Overall mortality was 3.1% (n = 7) for the AV group, 2.3% (n = 1) for the AR group, 0% for the V/A group, and 10.0% (n = 1) for the AA group. Complications included reoperation for bleeding in 10 (4.5%), two (4.7%), one (11.1%), and one (11.1%) for the four groups respectively; and sternal wound infection in eight (3.6%) patients of the AV group and one (2.3%) patient of the AR group. Five (2.3%) patients of the AV group suffered stroke. Isolated or more complicated aortic valve, root and ascending aortic surgery is feasible and safe through a minimally invasive approach with acceptable incidence of complications and mortality, without compromising the efficacy of the procedure. PMID- 11060584 TI - Closed chest totally endoscopic coronary artery bypass surgery: fantasy or reality? AB - With the introduction of the da Vinci robotic surgical system (Intuitive Surgical, Mountain View, CA) into minimally invasive cardiac surgery, the outlook of performing coronary artery bypass operations "closed chest" became a reality. Between May 1999 and July 2000 this wrist-enhanced instrumentation was used in 143 patients (107 men, 36 women, median age 63 10.3 y). Thirteen patients suffering from coronary artery disease (CAD) were treated as totally endoscopic coronary artery bypass (TECAB), 79 patients underwent a minimally invasive direct coronary artery bypass procedure, and 35 patients were treated using the robotic enhanced Dresden Technique. Preoperative survival was 100%. All patients in the TECAB group were operated upon via a three- or four-point stab incision using the da Vinci robot for internal mammary artery takedown and for performance of anastomoses. These new robotic-enhanced surgical techniques promote an optimistic way of thinking about the further development of these procedures and its application in patients suffering from CAD. PMID- 11060585 TI - Surgical alternatives to transplantation and assist devices in the treatment of heart failure. AB - It is estimated that in the United States 30,000 patients with end-stage heart disease are eligible for heart transplantation, although less than 2500 are actually performed each year. As the population ages, several thousand other patients who are ineligible for cardiac transplantation will be left with limited options. Surgical strategies aimed at halting the progression of heart failure have therefore been devised. High-risk coronary artery bypass grafting can improve ejection fraction and alleviate heart failure symptoms in appropriately selected patients. Mitral valve repair and ventricular aneurysm resection/plication procedures may improve heart failure by decreasing the volume load on the left ventricle. Myoreduction operations (Batista operation) aim to improve the volume and wall stress relationship of the dilated left ventricle, but their clinical application has remained experimental. PMID- 11060586 TI - Minimal access reoperative mitral and aortic valve surgery. AB - Minimally invasive cardiac surgery has allowed surgeons to perform valve procedures with a morbidity and mortality comparable with conventional resternotomy approaches while reducing postoperative ventilatory and intensive care unit requirements and overall hospital length of stay. Additionally, patient satisfaction with rapid recovery, earlier return to work, and improved cosmetic results has pushed the pendulum of reoperative valve surgery towards minimally invasive techniques. We reviewed our institutional data consisting of 129 patients requiring reoperative valve surgery over the past 4 years, which was accomplished using these minimally invasive approaches. PMID- 11060587 TI - Coronary anastomotic devices: theory and patented ideas on micromechanical fastening. AB - Coronary anastomotic devices are being designed to reduce the laborious, complex suturing approach to endoscopic coronary surgery. An anastomotic device should be safe and reliable, it should allow full view of the vessel parts to be bonded, it must provide a simple and rapid deployment, and should be hemodynamically adequate. Three anastomotic device categories found in the (patent) literature are discussed that use micromechanical fastening techniques. First, devices using individual bonding elements; second, devices using bonding elements anchored to extra-luminal frames; third, devices using an internal frame, often a stent-like structure. Anastomotic devices described in the (patent) literature to date fail to meet all requirements for endoscopic coronary application. PMID- 11060588 TI - The cancer patient with chronic pain due to herpes zoster. AB - Postherpetic neuralgia (PHN) is the most common complication of herpes zoster, and as such has been an area of extensive medical research for the past three decades. The patients at highest risk for PHN include those older than 50 years, those with severe acute cases of zoster, and those with shingles in a trigeminal distribution. As persons with malignancy are at a high risk for developing zoster itself, PHN is a complication that will be faced by many of these patients and their caregivers. This article reviews the available treatments and preventative measures for this debilitating condition. PMID- 11060589 TI - The role of radiofrequency in the management of complex regional pain syndrome. AB - The nomenclature, pathophysiology, and treatment modalities of complex regional pain syndrome (CRPS) are controversial. Thus far there are no specific, scientifically valid treatments for the management of CRPS. The numerous modalities of treatment range from sympathetic ganglion blocks, intravenous regional blocks, administration of a multitude of pharmacologic agents and behavioral interventions, to surgical sympathectomy. Minimally invasive radiofrequency lesioning for managing CRPS is a modality in its developmental stages. This article describes radiofrequency lesioning techniques in managing CRPS. PMID- 11060590 TI - Genetic therapy for pain management. AB - Two approaches to genetic therapy for the management of chronic pain have recently been investigated in animal models of pain. First, transgene-mediated delivery of antinociceptive molecules to the cerebrospinal fluid has been performed with engineered cell lines transplanted to the subarachnoid space and with recombinant adenoviruses that transduce pia mater cells. Second, the phenotype of nociceptive neurons has been altered by recombinant herpes viruses overexpressing antinociceptive peptides or reducing expression of endogenous nociceptive molecules. Both approaches attenuate or reverse persistent nociceptive states, suggesting use in the development of genetic therapy for pain management in humans. PMID- 11060591 TI - Nociceptors for the 21st century. AB - This review summarizes recent developments in the context of the neurochemical classification of nociceptors and explores the relationships between functionally and neurochemically defined subgroups. Although the complete picture is not yet available, several lines of intriguing evidence suggest that despite the complexity and diversity of nociceptor properties, a relatively "simple" neurochemical classification fits well with several recently identified molecular characteristics. PMID- 11060592 TI - The role of nitric oxide in nociception. AB - Pharmacologic, electrophysiologic, and immunohistochemical studies have suggested a role of nitric oxide (NO) in nociception processing. Recent studies have indicated that NO may modulate spinal and sensory neuron excitability through multiple mechanisms that may underlie its distinctive roles in different pain states. Differential regulation of a family of NO-producing enzymes, NO synthases, contributes mainly to the complexity underlying the role of NO in nociception. This review summarizes the latest advances in our understanding of the contribution of NO to pain transduction. Possible cellular mechanisms regarding the connection between NO production and the abnormal sensation derived from different stimuli and pathologic conditions are discussed. PMID- 11060593 TI - Exploring the pain "neuromatrix". AB - A considerable number of functional imaging studies have demonstrated the involvement of multiple central regions during the experience of pain. These regions process information in circuits that can broadly be assumed to process the affective, sensory, cognitive, motor, inhibitory, and autonomic responses stimulated by a noxious event. The concept of a "neuromatrix" for pain processing is, therefore, well supported. There is, however, scant evidence for any particular regional or circuit dysfunction during clinical pain. To be clinically useful, functional imaging may have to step beyond the generalities of the neuromatrix. PMID- 11060594 TI - Visceral nociception. AB - Visceral pain is of great concern to the medical community because it remains particularly resistant to current clinical treatments. A serendipitous and initially unexplainable clinical finding that a punctate midline dorsal column lesion is effective in eliminating visceral pain, however, has initiated a resurgence of interest in the study of the basic mechanisms of visceral nociception. Clinical and anatomic findings have determined that visceral pain either of thoracic or pelvic origin can be relieved by carefully placed lesions directed at the lateral edge or the medial edge of the gracile fasciculus, respectively. Studies are demonstrating that visceral pain is quite unique from cutaneous pain. PMID- 11060595 TI - Calcium channel blockers and pain therapy. AB - This review focuses on the advances in the development of N-type calcium channel blockers as analgesic agents over the last 2 years. Firstly, it highlights the clinical progress with SNX-111 (Ziconotide; Elan Pharmaceuticals, Smithfield, RI) and then secondly, it outlines the various approaches being taken by researchers to design orally active, selective, small molecule modulators without the perceived disadvantages associated with SNX-111. PMID- 11060596 TI - Visceral pain. AB - Visceral pain, although different from somatic pain in several important features, is not as widely researched and consequently our knowledge of neurophysiologic mechanisms as well as clinical management of visceral pain states remains unsatisfactory. Several recent studies have employed different visceral pain animal models to provide insight into the peripheral and central nervous system mechanisms underlying pain originating from the urinary bladder, ureter, and gastrointestinal tract. The effects of opioid and nonopioid drugs in these models have also been evaluated and are reviewed in this article. The importance of anatomic pathways relaying pain sensation in the central nervous system, particularly the newly described dorsal column pathway, is also discussed. In human subjects, new techniques like positron emission tomography are now being used to better understand visceral pain perception. Such findings deriving from basic animal research and human studies summarized in the present overview lead to a better understanding of visceral pain states and may be helpful in developing better treatment strategies to combat visceral pain states in the clinical setting. PMID- 11060597 TI - Strong confinement and oscillations in two-component bose-einstein condensates AB - We present a new model of Bose-Einstein condensate dynamics based on strong confinement near the ground state. The model is based on a combined particle-wave view of the condensate and predicts oscillations in a two-component condensate, based on interference of nonspreading wave packets moving within a pair of tilted nearly square potentials. The oscillations are similar to those recently reported for a magnetically trapped 87Rb condensate, and the model's predictions give good quantitative agreement with the experiments. PMID- 11060598 TI - Effect of noise on a quantum bound state AB - The evolution of a quantum wave function on a lattice with time-dependent random disorder is studied. It is found that the bound state wave function of a quantum system exhibits intrinsic instability with respect to noisy disorder in the statistical sense. Under the general scheme of time coarse graining, we show nonperturbatively that the stationary attractive potential is completely washed out at large time scales by the existence of the noisy disorder to any order of density correlations. PMID- 11060599 TI - New constraints on neutrino physics from BOOMERANG data AB - We have performed a likelihood analysis of the recent data on the cosmic microwave background radiation anisotropy from the BOOMERANG experiment. These data place a strong upper bound on the radiation density present at recombination. Expressed in terms of the equivalent number of neutrino species the 2sigma bound is N(nu)>Omega(i). Nonlinearly, these waves lead to multiscale spatially coherent structures, substantial cross-field transport, ion energization, and phase-space diffusion. Large spikes are formed in the parallel electric field time series. These signatures are similar to the Fast Auroral Snapshot satellite observations in the upward current region. PMID- 11060620 TI - Extended X-Ray absorption fine structure from hydrogen atoms in water AB - We report the first quantitative measurement of extended x-ray absorption fine structure (EXAFS) from hydrogen atoms. A single oscillation is observed from gaseous water consistent with the location of the covalently bonded hydrogen in H 2O. The experimental phase and amplitude of the oscillation are in excellent agreement with curved wave multiple scattering calculations for isolated water molecules. With this determination of the O-H scattering phase shift we have quantified the covalent hydrogen bond distance (0.95+/-0.03 A) in liquid water, thus demonstrating that hydrogen EXAFS can become a valuable complement to existing structural methods in chemistry and biology. PMID- 11060621 TI - Dynamic heterogeneity of relaxations in glasses and liquids AB - We report an investigation of the heterogeneity in supercooled liquids and glasses using the non-Gaussianity parameter. We simulate selenium and a binary Lennard-Jones system by molecular dynamics. In the non-Gaussianity three time domains can be distinguished: an increase on the ps scale due to the vibrational (ballistic) motion of the atoms, followed by a growth, due to local relaxations ( beta relaxation) at not too high temperatures, and finally a slow drop at long times. The non-Gaussianity follows in the intermediate time domain a sqrt[t] law. This is explained by collective hopping and dynamic heterogeneity. We support this finding by a model calculation. PMID- 11060622 TI - Diffusion coefficients in a lamellar lyotropic phase: evidence for defects connecting the surfactant structure AB - We measure diffusion coefficients in the lamellar phase of the nonionic binary system C(12)EO(6)/H(2)O using fluorescence recovery after photobleaching. The diffusion coefficient across the lamellae shows an abrupt increase upon approaching the lamellar-isotropic phase transition. We interpret this feature in terms of defects connecting the surfactant structure. An estimation of the defect density and of the variation in defect energy close to the transition is given in terms of a simple model. PMID- 11060623 TI - Anisotropic nonlinear elastic properties of an icosahedral quasicrystal AB - We show that the nonlinear behavior of transverse acoustic waves can reveal the anisotropic structure of an icosahedral quasicrystal, at the macroscopic level. We report experiments performed in i-Al-Pd-Mn. We observe that a primary transverse acoustic wave can generate a second harmonic transverse acoustic wave. We also observe a specific relation between the polarization directions of those waves. These observations are manifestations on a macroscopic scale of the long range order in quasicrystals. PMID- 11060624 TI - Conformations of random polyampholytes AB - We study the size R(g) of random polyampholytes (i.e., polymers with randomly charged monomers) as a function of their length N. All results of our extensive Monte Carlo simulations can be rationalized in terms of the scaling theory we develop for the Kantor-Kardar necklace model, although this theory neglects the quenched disorder in the charge sequence along the chain. We find approximately N1/2. The elongated globule model, the initial predictions of both Higgs and Joanny ( approximately N1/3) and Kantor and Kardar ( approximately N), and previous numerical estimates are ruled out. PMID- 11060625 TI - A discotic disguised as a smectic: A hybrid columnar bragg glass AB - We show that discotics, lying deep in the columnar phase, can exhibit an x-ray scattering pattern which mimics that of a somewhat unusual smectic liquid crystal. This exotic, new glassy phase of columnar liquid crystals, which we call a "hybrid columnar Bragg glass," can be achieved by confining a columnar liquid crystal in an anisotropic random environment of, e.g., strained aerogel. Long ranged orientational order in this phase makes single-domain x-ray scattering possible, from which a wealth of information could be extracted. We give detailed quantitative predictions for the scattering pattern in addition to exponents characterizing anomalous elasticity of the system. PMID- 11060626 TI - Critical behavior of the de gennes elastic constants near the nematic-smectic- A transition of TBBA AB - We have studied the behavior of elastic constants A, B, and C deduced from ultrasound velocity anisotropies in the vicinity of the nematic-smectic- A transition of terephthal-bis- p- p(')-butylaniline. A is associated with compressibility, B with layer compression, and C with the coupling between compressibility and layer compression. We show that the exponent of A is of the preasymptotic 3D- XY type, whereas those of B and C are in between the 3D- XY values and those associated with the anisotropic fixed point. This behavior is consistent with the extended crossover regime predicted by Patton and Andereck [Phys. Rev. Lett. 69, 1556 (1992)]. PMID- 11060627 TI - Supercooled confined water and the mode coupling crossover temperature AB - We present a molecular dynamics study of the single-particle dynamics of supercooled water confined in a silica pore. Two dynamical regimes are found. Close to the hydrophilic substrate molecules are below the mode coupling crossover temperature, T(C), already at ambient temperature. The water closer to the center of the pore (free water) approaches upon supercooling T(C) as predicted by mode coupling theories. For free water the crossover temperature and crossover exponent gamma are extracted from power-law fits to both the diffusion coefficient and the relaxation time of the late alpha region. PMID- 11060628 TI - Primary and secondary nucleation of the transition between the A and B phases of superfluid 3He AB - We have studied nucleation in superfluid 3He across the A-B phase transition driven by a magnetic field, in a controllable environment at very low temperatures. Both B-->A and A-->B secondary nucleation appear to be governed by the survival of pockets of the new phase trapped at surfaces. We find that, at fields near B(AB), primary A-->B nucleation cannot be triggered by ionizing or neutron irradiation even at very high intensities. In our cell primary A-->B nucleation can only be triggered externally by mild mechanical shock. PMID- 11060629 TI - High-frequency acoustics of 3He in aerogel AB - High-frequency ( approximately 15 MHz) acoustics were performed on 3He in 98% porous silica aerogel using an acoustic cavity technique. Measurements of the sound attenuation in the normal Fermi liquid and superfluid display behavior quite different from the bulk owing to strong elastic scattering of quasiparticles. The transition from first-to-zero sound is completely obscured with a quasiparticle mean-free path estimated to be in the range of 200-300 nm. No collective mode attenuation peak was observed at or below the superfluid transition. PMID- 11060630 TI - Meniscus instability in a thin elastic film AB - A new kind of meniscus instability leading to the formation of stationary fingers with a well-defined spacing has been observed in experiments with elastomeric films confined between a plane rigid glass and a thin curved glass plate. The wavelength of the instability increases linearly with the thickness of the confined film, but it is remarkably insensitive to the compliance and the energetics of the system. However, lateral amplitude (length) of the fingers depends on the compliance of the system and on the radius of curvature of the glass plate. A simple linear stability analysis is used to explain the underlying physics and the key observed features of the instability. PMID- 11060631 TI - Theoretical analysis of the growth mode for thin metallic films on oxide substrates AB - We show how the growth mode of a thin metallic film on an insulating substrate can be predicted theoretically by combining thermodynamic considerations with ab initio calculations for ordered metal/insulator interfaces at low coverage. Our approach is illustrated by calculations for Ag film deposited on an MgO substrate. Ab initio calculations predict high mobility of adsorbed Ag atoms on MgO, even at low temperatures, which greatly aids their aggregation. PMID- 11060632 TI - Exploiting the difference in lattice structures for formation of self-assembled PbS dots on InP(110) AB - Via the example of PbS self-assembled nanodots on InP(110) we show that spontaneous self-organization at semiconductor interfaces can be achieved due to different lattice structure of the constituents with close lattice constants. Regularly shaped triangular pyramidal PbS dots almost perfectly oriented on the substrate and distributed reasonably uniform in size can be grown in this way by molecular beam epitaxy. The average lateral dot size can be varied between 15 and 35 nm by appropriate choice of growth temperature and total PbS coverage. PMID- 11060633 TI - Direct observation of vibrational energy delocalization on surfaces: CO on Ru(001) AB - We report the experimental observation of the gradual transition from a local oscillator to a two-dimensional delocalized phonon, observed for the CO-stretch vibration of carbon monoxide adsorbed on a Ru(001) surface by means of broadband infrared saturation sum-frequency spectroscopy. The data are theoretically reproduced by an exchange model with residence times of the excitation down to 2.5 ps. PMID- 11060634 TI - Spontaneous deformation of the fermi surface due to strong correlation in the two dimensional t- J model AB - The Fermi surface of the two-dimensional t- J model is studied using the variational Monte Carlo method. We study the Gutzwiller-projected d-wave superconducting state with an additional variational parameter t(')(v) corresponding to the next-nearest-neighbor hopping term. It is found that the finite t(')(v)<0 gives the lowest variational energy in the wide range of hole doping rates. The obtained momentum distribution function shows that the Fermi surface deforms spontaneously. It is also shown that the Van Hove singularity is always located very close to the Fermi energy. Using the Gutzwiller approximation, we show that this deformation is due to the Gutzwiller projection operator or the strong correlation. PMID- 11060635 TI - X-Ray anomalous scattering study of a charge-ordered state in NaV2O5 AB - Charge ordering of V4+ and V5+ in NaV2O5 has been studied by an x-ray diffraction technique using anomalous scattering near a vanadium K-absorption edge to critically enhance a contrast between the two ions. A dramatic energy dependence of the superlattice intensities is observed below T(C) = 35 K. The charge ordering pattern is the fully charged zigzag-type ladder with the unit cell 2ax2bx4c, but not the chain-type originally proposed for the spin-Peierls state. Charge disproportionation suggested in our model as the average valence V(4.5+/ delta(c)/2) is observed below T(C), showing continuous variation of delta(c) as a function of temperature. PMID- 11060636 TI - Independent freezing of charge and spin dynamics in La1.5Sr0.5CoO4 AB - We present elastic and quasielastic neutron scattering measurements characterizing peculiar short-range charge-orbital and spin order in the layered perovskite material La1.5Sr0.5CoO4. We find that below T(c) approximately 750 K holes introduced by Sr doping lose mobility and enter a statically ordered charge glass phase with loosely correlated checkerboard arrangement of empty and occupied d(3z(2)-r(2)) orbitals ( Co3+ and Co2+). The dynamics of the resultant mixed spin system is governed by the anisotropic nature of the crystal-field Hamiltonian and the peculiar exchange pattern produced by the orbital order. It undergoes a spin freezing transition at a much lower temperature, T(s) less, similar30 K. PMID- 11060637 TI - Analysis of the metallic phase of two-dimensional holes in SiGe in terms of temperature dependent screening AB - We find that temperature dependent screening can quantitatively explain the metallic behavior of the resistivity on the metallic side of the so-called metal insulator transition in p-SiGe. Interference and interaction effects exhibit the usual insulating behavior which is expected to overpower the metallic background at sufficiently low temperatures. We find empirically that the concept of a Fermi liquid describes our system with its large interaction parameter r(s) approximately 8. PMID- 11060638 TI - Is the intrinsic thermoelectric power of carbon nanotubes positive? AB - The thermoelectric power (TEP) of single-walled carbon nanotubes (SWNTs) is extremely sensitive to gas exposure history. Samples exposed to air or oxygen have an always positive TEP, suggestive of holelike carriers. However, at fixed temperature the TEP crosses zero and becomes progressively more negative as the SWNTs are stripped of oxygen. The time constant for oxygen adsorption/desorption is strongly temperature dependent and ranges from seconds to many days, leading to apparently "variable" TEP for a given sample at a given temperature. The saturated TEP can be accounted for within a model of strong oxygen doping of the semiconducting nanotubes. PMID- 11060639 TI - Detecting electronic states at stacking faults in magnetic thin films by tunneling spectroscopy AB - Co islands grown on Cu(111) with a stacking fault at the interface present a conductance in the empty electronic states larger than the Co islands that follow the stacking sequence of the Cu substrate. Electrons can be more easily injected into these faulted interfaces, providing a way to enhance transmission in future spintronic devices. The electronic states associated with the stacking fault are visualized by tunneling spectroscopy, and its origin is identified by band structure calculations. PMID- 11060640 TI - Anomalous thermopower in the metalliclike phase of a 2D hole system AB - We report very low temperature ( T) thermopower and resistivity ( rho) measurements on variable-density, two-dimensional hole systems confined to GaAs quantum wells. As the hole density is lowered from 1.49x10(11) cm(-2) to 0.14x10(11) cm(-2), the system crosses from an insulating ( drho / dT less, similar0) to a metallic regime ( drho / dT>0) and finally displays insulating behavior ( drho / dT<0). Diffusion thermopower shows a striking sign reversal in a narrow range of density in the metallic regime, suggesting a qualitative change in the conduction or the scattering mechanism. PMID- 11060641 TI - Strong-coupling expansions for multiparticle excitations: continuum and bound states AB - We present a new linked cluster expansion for calculating properties of multiparticle excitation spectra to high orders. We use it to obtain the two particle spectra for systems of coupled spin-half dimers. We find that even for weakly coupled dimers the spectrum is very rich, consisting of many bound states. The number of bound states depends on both geometry of coupling and frustration. Many of the bound states can only be seen by going to sufficiently high orders in the perturbation theory, showing the extended character of the pair attraction. PMID- 11060642 TI - Observation of quantum coherence in mesoscopic molecular magnets AB - By applying a transverse magnetic field B( perpendicular) of sufficient strength to the uniaxial molecular magnets Fe8 and Mn12, the tunneling splitting Delta(t) of their S = +/-10 magnetic ground states can be made large compared to perturbations such as hyperfine and dipolar interactions. We present evidence for such a Delta(t) from magnetic specific heat data below 1 K that is consistent with coherent quantum mechanical tunneling in a "mesoscopic" system under such conditions. PMID- 11060643 TI - Optical anisotropy of the SiC((001))- (3x2) surface: evidence for the two-adlayer asymmetric-dimer model AB - The structure of the ( 3x2) reconstruction of beta-SiC(001) surface has been identified by comparing reflectance anisotropy spectra calculated from first principles with recent measurements. Only the calculations for the two-adlayer asymmetric-dimer model agree with experiment. The two prominent peaks at 3.6 and 5.0 eV found experimentally are assigned to electronic transitions between surface and bulklike electronic states. A further pronounced anisotropy at 2.0 eV, due to transitions between surface states, is predicted. PMID- 11060644 TI - First observation of nonreciprocal X-Ray gyrotropy AB - We report the first observation of a nonreciprocal x-ray linear dichroism caused by the time-reversal odd, real part zeta of the complex gyrotropy tensor zeta(*) which is dominated by electric dipole-electric quadrupole E1E2 interference terms. A nonreciprocal transverse anisotropy was observed in the low temperature insulating phase of a Cr doped V2O3 Mott crystal when a single antiferromagnetic domain was grown by magnetoelectric annealing along the hexagonal c axis. This new element (edge) specific spectroscopy could nicely complement x-ray magnetic circular dichroism which is silent for antiferromagnetic materials. PMID- 11060645 TI - Salt-induced DNA-histone complexation. AB - We study numerically the binding of one semiflexible charged polymer onto an oppositely charged sphere. Using parameters appropriate for DNA-histone complexes, we find complete wrapping for intermediate salt concentrations only, in agreement with experiments. For high salt concentrations, a strongly discontinuous dewrapping occurs. For low salt concentrations, we find multiple conformational transitions, leading to an extended DNA configuration. The wrapped states are characterized by spontaneously broken rotational and mirror symmetries, giving rise to four distinct structures. PMID- 11060646 TI - Fluctuation-facilitated charge migration along DNA. AB - We propose a model Hamiltonian for charge transfer along the DNA double helix with temperature-driven fluctuations in the base pair positions acting as the rate limiting factor for charge transfer between neighboring base pairs. We compare the predictions of the model with the recent work of Barton and Zewail on the unusual two-stage charge transfer of DNA. PMID- 11060647 TI - Dilution wave and negative-order crystallization kinetics of chain molecules. AB - We show that the crystal growth rate of a very long-chain n-alkane C C(198)H(398) from solution can decrease with increasing supersaturation and follow strongly negative order kinetics. The experimental behavior can be well represented by a theoretical model which allows the molecule to attach and detach as either extended or folded in two. The obstruction of extended-chain growth by unstable folded depositions increases disproportionately with increasing concentration. As a consequence of this abnormal kinetics, a "dilution wave" can propagate and trigger a folded-to-extended-chain transformation on its way. PMID- 11060649 TI - Comment on "Astrophysical jets as exact plasma equilibria" PMID- 11060648 TI - Universal aspects of the chemomechanical coupling for molecular motors. AB - The directed movement of molecular motors is studied theoretically within a general class of nonuniform ratchet models in which the motor can attain M internal states and undergo transitions between these states at K spatial locations. The functional relationship between the motor velocity and the concentration of the fuel molecule is analyzed for arbitrary values of M and K. This relationship is found to exhibit universal features which depend on the number of unbalanced transitions per motor cycle arising from the enzymatic motor activity. This agrees with experimental results on dimeric kinesin and is predicted to apply to other cytoskeletal motors. PMID- 11060650 TI - Bogoyavlenskij replies: PMID- 11060651 TI - Comment on "Hydrodynamic coarsening of binary fluids" PMID- 11060652 TI - Solis, olvera de la cruz, and smith reply: PMID- 11060653 TI - Comment on "Degenerate wannier theory for multiple ionization" PMID- 11060654 TI - Pattard and rost reply: PMID- 11060655 TI - Comment on "New approach in equilibrium theory for strained layer relaxation" PMID- 11060656 TI - Fischer replies: PMID- 11060657 TI - Novel therapy for COPD. AB - The incidence of chronic obstructive pulmonary disease (COPD) is increasing throughout the world. Much less is known about the pathogenesis of COPD than that of asthma and there is little response to current therapy. Most patients with COPD have acquired their lung disease through smoking cigarettes, and the major step in management is to minimise further damage by stopping this habit. A number of therapies are being developed for the treatment of COPD; including new bronchodilators such as tiotropium bromide, agents to block inflammation induced by neutrophils and macrophages, as well as strategies to combat proteases and oxidants. The long-term goal is to provide therapy that retards the accelerated loss of lung function occurring in COPD. Development of novel therapies for COPD requires reliable Phase II decision making before entering large scale Phase III studies. The patient with COPD is often overlooked compared to their asthmatic counterpart, who benefit from an urgent need to identify novel targets and better therapy. PMID- 11060658 TI - Novel therapy for asthma. AB - The health burden of asthma is increasing globally at an alarming rate, providing a strong impetus for the development of new therapeutics. Currently available inhaled bronchodilators and anti-inflammatory drugs are effective in most asthmatics, but this palliative therapy requires long-term daily administration. Despite considerable efforts by the pharmaceutical industry, it has been difficult to develop novel therapeutic agents; the leukotriene antagonists and synthesis inhibitors being the only new class of asthma treatments to have been licensed in the last 30 years. It is clearly important to understand more about the underlying mechanisms of asthma and about how current drugs work before rational improvements in therapy can be expected. There are numerous therapies in clinical development that combat the inflammation found in asthma, specifically targeting eosinophils, IgE, adhesion molecules, cytokines and chemokines, inflammatory mediators and cell signalling. In particular, there is the obvious need for new therapy for severe asthma that is poorly controlled by high doses of corticosteroids, as well as agents to counter acute emergency asthma. A long-term goal is to develop disease-modifying immunotherapy, that could be introduced in childhood to alter the natural history of asthma. Thanks to the extensive efforts of the pharmaceutical industry, in the near future we can expect the introduction of a range of novel therapies for asthma. PMID- 11060659 TI - CCR3 blockade as a new therapy for asthma. AB - Among the inflammatory cells infiltrating the lungs of asthmatic patients, eosinophils and Th2 cells are thought to play a central role in the pathogenesis of this disease. Several studies have implicated that chemokines are prime candidates for being responsible for the selective recruitment of the leukocyte subsets found in atopic diseases. Regulated upon activation, normal T-cell expressed and secreted (RANTES), monocyte chemoattractant protein-3 (MCP-3), MCP 4 and the eotaxins, for example, have been shown in vitro to potently induce eosinophil chemotaxis as well as initiate several other pro-inflammatory activities such as integrin activation, lipid mediator biosynthesis and degranulation. Ligand binding and chemotaxis experiments with these chemokines demonstrated that a G-protein coupled-receptor (GPCR) cloned from eosinophils, termed CCR3, was responsible for producing a chemokine selectivity profile identical to that of eosinophils. In addition, blocking CCR3 on eosinophils, with a monoclonal antibody, completely abolished eosinophil responses to these chemokines. Together these studies strongly suggest a central role for this receptor in eosinophil trafficking. CCR3 has also been found on in vitro derived Th2 cells and on T-cells co-localising with eosinophils in diseased tissue, thus revealing a possible pathogenic mechanism for T-cell recruitment into the airways. Therefore, blockade of CCR3 represents a highly attractive and innovative strategy for asthma therapy. PMID- 11060660 TI - Postnatal glucocorticosteroid therapy for treatment and prevention of neonatal chronic lung disease. AB - Neonatal chronic lung disease (CLD) is a persistent complication, primarily of premature infants. Postnatal glucocorticoid therapy is widely used in the treatment and prevention of CLD. Most studies reveal acute improvement in the pulmonary status of infants treated with postnatal glucocorticoid therapy. Recent studies of 'earlier' intervention (< 14 days of age) demonstrated a reduction in mortality and in the occurrence of CLD between 28 days of age and 36 weeks postmenstrual age. Great concern remains, however, regarding the potential adverse outcomes, including growth inhibition, infection, catastrophic GI complications and CNS injury. Therefore, the use of postnatal glucocorticoid therapy remains controversial with respect to the clinical indications for initiating therapy, the dose, duration, onset and route of administration, as well as potential benefits and risks. Inhaled glucocorticoid therapy is increasingly used to treat and prevent CLD in order to avoid adverse effects of high dose systemic glucocorticoid therapy. Recent studies with inhaled glucocorticoid therapy show promise. Further work, however, for improving aerosol delivery and deposition, will be needed to refine their role in the prevention and treatment of CLD. Future studies enabling early, accurate identification of infants at greatest risk for CLD, coupled with a more comprehensive understanding of the different pathogeneses, will provide information regarding appropriate timing of onset, dosing, route of therapy and duration of intervention. PMID- 11060661 TI - Ascomycins: promising agents for the treatment of inflammatory skin diseases. AB - Ascomycin derivatives represent a novel class of anti-inflammatory macrolactams currently under development for the treatment of skin diseases. The main biological effect of ascomycins is an inhibition of the synthesis of both Th1 and Th2-type cytokines in target cells. Several compounds are being developed with SDZ ASM 981 being at the most advanced stage. It has high anti-inflammatory activity in animal models of skin inflammation and does not induce skin atrophy. Topical application of SDZ ASM 981 was shown to be effective in atopic dermatitis (AD), allergic contact dermatitis and also in psoriasis under semi-occlusive conditions. In patients with AD, SDZ ASM 981 cream led to consistently low systemic exposure even when applied on large areas of skin. SDZ ASM 981 overcomes the drawbacks of current topical therapies of inflammatory skin diseases as its safety profile is better than that of topical corticosteroids. Studies continue to investigate its efficacy and safety in the treatment of inflammatory skin diseases. PMID- 11060662 TI - New drugs in the treatment of psoriasis. AB - Psoriasis is one of the most common skin disorders affecting approximately 2% of the population; the disease is recurrent and can be very debilitating. The cause of psoriasis is unknown, although it appears to be an autoimmune disease with a genetic component to its aetiology. Past topical treatments such as emollients, coal tar and dithranol have been messy, cosmetically unacceptable or of low efficacy, while older systemic therapies have suffered from significant side effects. Newer drugs with better therapeutic indexes and new antiproliferative/immunomodulatory therapies based on an increased understanding of the origins of psoriasis have brought us closer to the goal of safely and efficaciously treating the disease. This review will cover the newest topical and systemic drugs currently in use, in clinical trials or preclinical development. PMID- 11060663 TI - The treatment of psoriasis with IL-10: rationale and review of the first clinical trials. AB - By virtue of its anti-inflammatory and immunosuppressive properties, IL-10 plays a crucial role in several immune reactions, including regulatory mechanisms in the skin. In psoriasis, a common cutaneous immune disease, a relative deficiency in cutaneous IL-10 expression is observed. Several lines of evidence suggest that IL-10 could have antipsoriatic abilities. One pilot and two Phase II trials with sc. IL-10 administration over 3 - 7 weeks in patients with moderate to severe psoriasis have supported this hypothesis. The therapy was well-tolerated and clinical efficiency was found in the majority of patients. Immunosuppressive effects (depressed monocytic HLA-DR expression, TNF-alpha and IL-12 secretion capacity, IL-12 plasma levels and responsiveness to recall antigens) as well as a shift towards a Type 2 cytokine pattern (increasing proportion of IL-4, IL-5, and IL-10 producing T-cells, selective increase in IgE serum levels) were observed. These investigations suggest that IL-10 is of major importance in psoriasis and show that IL-10 administration represents a new therapeutic approach. However, long-term administration of large recombinant protein limits the value of this novel therapeutic approach. As such, neither oral nor topical applications are possible; there is a risk of the development of neutralising antibodies. PMID- 11060664 TI - Anti-TNF agents in Crohn's disease. AB - The current treatment of Crohn's disease is limited by a lack of long-term efficacy of corticosteroid therapy and the associated side effects. Biological treatment strategies aimed at neutralising immune responses, offer new opportunities for the management of chronic inflammatory disorders. In Crohn's disease, anti-TNF agents have taken the lead in development of immune-modulating drugs since TNF is known to be a pivotal cytokine in this illness. Different strategies have been explored aimed at inhibiting TNF but at present, the majority of clinical data have been obtained with monoclonal antihuman TNF antibodies. The chimeric anti-TNF IgG1 antibody infliximab (cA2, Remicade, Centocor) has been proven, in multiple clinical trials, to be an effective and well tolerated therapy for the management of acute Crohn's disease and recently this compound has obtained FDA and European Medicines Evaluation Agency approval. Although there are some concerns about immunogenicity of the anti-TNF antibody resulting in the formation of human antichimeric antibodies (HACA) as well as lymphoproliferative disorders, the clinical benefit in the treatment of refractory Crohn's disease is a major therapeutic breakthrough. Further studies will be needed to establish the role and optimal dosing scheme of anti-TNF antibodies in maintenance of remission, monitor safety in the long run and to evaluate the effectiveness of alternative anti-TNF agents such as the TNF receptor/Fc fusion protein etanercept (Enbrel, Immunex) and TNF synthesis inhibitor thalidomide. PMID- 11060665 TI - IL-1 inhibitors: novel agents in the treatment of rheumatoid arthritis. AB - IL-1 is a pleiotropic cytokine shown to play a major role in synovitis and in the mechanisms that lead to the progressive joint destruction of rheumatoid arthritis (RA). IL-1 receptor antagonist (IL-1Ra), a member of the IL-1 family, binds IL-1 receptors but does not induce a cellular response. IL-1Ra competitively inhibits the binding of IL-1 to its cell surface receptors and thus acts as an endogenous anti-inflammatory mediator. In different experimental animal models of arthritis systemic administration of IL-1Ra, or local delivery into the joints by gene therapy attenuated the severity of the inflammatory response and reduced articular destruction. In addition, treatment of RA patients with IL-1Ra led to an improvement in different clinical and biological parameters and to a reduction in the radiological signs of joint erosions. Recently, interesting results were obtained using IL-1Ra in combination with methotrexate, a well-known antirheumatic drug, or in combination with other strategies designed to block the effects of tumour necrosis factor (TNF)-alpha. Encouraging results also have been reported in both in vitro and in vivo experimental models of arthritis by using other strategies designed to block the effects of IL-1. PMID- 11060666 TI - Therapeutic and chemical developments of cholecystokinin receptor ligands. AB - Cholecystokinin (CCK) is an important 'brain-gut' hormone located both in the gastrointestinal (GI) system and in the CNS. At least two different G-coupled high affinity receptors have been identified: the CCK-A and the CCK-B receptors. Although the complex biological role of CCK is, as yet, not fully understood, its connection with many different physiological processes both at the GI level and at the CNS level is now well established. There is much potential for therapeutic use of CCK receptor ligands, however, clear investigations have yet to be completed. Several chemical families have been investigated over the last 20 years to find potent, subtype selective and stable CCK receptor agonists and antagonists. The main goal was to discover new therapeutic drugs acting on GI and/or on CNS diseases and also, to obtain powerful pharmacological tools that could permit a better understanding of the biological role of CCK. Despite promising results from investigations into medicinal chemistry of CCK receptor ligands, the therapeutical applications of these ligands still remains to be defined. This article reviews the main biological role of CCK, the therapeutic potential of CCK-A and CCK-B receptor agonists and antagonists and the common compounds from the different families of ligands. PMID- 11060667 TI - Pharmacology and clinical experience with alosetron. AB - Alosetron (Lotronex) is a potent, highly selective 5-HT(3) antagonist. Animal models have shown it to be active in anxiety, psychosis, cognitive impairment, emesis and drug withdrawal, though its application in humans has been almost entirely restricted to irritable bowel syndrome (IBS). Alosetron does not cause adverse pharmacodynamic effects, is absorbed rapidly after oral administration and is widely distributed throughout tissues after oral or iv. dosing in animals. Its metabolism is rapid and extensive with N-demethylation, hydroxylation and oxidation. The drug, or its two principal metabolites, is equally excreted through the biliary tract and kidneys. Alosetron has proved safe in a range of toxicity studies; at high repeated dosing, clinical signs were transient and repeated administration produced no significant adverse effects on fertility, reproductive performance or fetal development. In pharmacokinetic studies, bioavailability of alosetron in healthy volunteers is approximately 60% and the plasma half-life is about 1.5 h. There are some gender differences in the pharmacokinetic profile, with 30 - 50% higher alosetron concentrations in females. No consistent differences in alosetron serum concentrations between the young and elderly were observed. The pharmacokinetics of single, oral doses of alosetron are linear up to 8 mg. In human pharmacodynamic studies, alosetron increased basal jejunal water and electrolyte absorption, increased colonic transit time and, consequently, whole gut transit time. Alosetron has been evaluated in two large Phase II trials (randomised, double-blinded, placebo controlled) and in Phase III trials which included a four-week observation period after cessation. Dose response studies suggested that the effective dosages could be between 1 and 2 mg, twice-daily. In Phase II trials, alosetron, 1 mg b.i.d., resulted in a greater proportion of non-constipated IBS patients reporting adequate relief of pain and discomfort, as well as improvement of bowel symptoms, frequency, urgency and stool consistency when compared with placebo. However, this beneficial effect was seen exclusively among females. Phase III studies evaluated exclusively females with non-constipated IBS and confirmed the results of the Phase II studies. Alosetron was well-tolerated in all studies, with the most frequently recorded adverse event being constipation. Thus, alosetron appears promising in the treatment of abdominal pain and discomfort and normalising of bowel function in patients with non-constipated IBS. It also improves quality of life, has a high degree of tolerability and has an excellent safety profile to date. PMID- 11060668 TI - Cerivastatin: the low-dose HMG-COA reductase inhibitor. PMID- 11060669 TI - Drugs in development for the treatment of multiple sclerosis: antigen non specific therapies--an update. AB - In the past few years there has been significant progress in the development of therapy for the treatment of relapsing remitting and secondary progressive multiple sclerosis. Research interest in multiple sclerosis (MS) therapeutics has remained high and clinical investigation into potential new therapies continues. This review summarises the advances with currently available therapies and briefly outlines the results from studies with other drugs being developed for the treatment of multiple sclerosis. PMID- 11060670 TI - HIV eradication revisited. AB - In the last three years, most efforts of virologists and immunologists involved in the field of HIV have been focussed on strategies aimed at eradicating the infection. So far, however, all attempts have been unsuccessful. Indeed, even if the currently available potent antiretroviral regimens are able to induce a profound and durable suppression of viral replication, they have poor effect on a viral reservoir of latently infected CD4+ T lymphocytes that rapidly reactivate after treatment discontinuation. Different approaches, including combined (immune based and antiretroviral) therapies, are therefore under investigation. The goal is to achieve an equilibrium where virus-induced immune dysfunction is prevented by an effective anti-HIV immune response. PMID- 11060671 TI - Interference with complement regulatory molecules as a possible therapeutic strategy in HIV infection. AB - Drugs which inhibit different stages of the HIV infection process, such as cell entry through CD4 and chemokine receptors, production of double stranded DNA from the HIV genome and maturation of newly produced viruses, are now proposed for AIDS therapy. None of these treatments, however, solve the problem of complete HIV eradication and the frequent appearance of mutants displaying drug resistance. We have recently detailed a strategy describing how HIV protects itself from the human complement and propose that interference of this resistance could be a possible target for therapy. PMID- 11060672 TI - Therapeutic developments in cytomegalovirus retinitis. AB - The incidence of cytomegalovirus (CMV) retinitis in AIDS has declined significantly due to the use of highly active antiretroviral therapy (HAART). However, patients with HIV, especially those failing HAART, may still suffer with CMV retinitis, which can lead to significant loss of vision and blindness. Ganciclovir has traditionally been considered the recommended treatment for CMV retinitis; however, due to side effects and the possibility of developing viral resistance, other agents may be preferred in certain situations. Foscarnet, which has similar efficacy to ganciclovir but a different side effect profile, is more difficult to administer and is less well-tolerated. Intravenous cidofovir, which may be more effective than either iv. ganciclovir or foscarnet, can also be used as a first line agent; however, it is associated with toxicity (renal and ocular) and thus needs careful use. Local therapy for CMV retinitis has been a significant advance. The intraocular ganciclovir implant has the highest efficacy of the approved agents and is well-tolerated. Fomivirsen, an oligonucleotide injected intravitreally, is a newly approved agent which offers alternative treatment. Intravitreal ganciclovir or foscarnet, although not approved, have been used successfully in some patients especially those with recurrent or refractory disease. The development of new anti-CMV agents has been stalled by the decreased incidence of the disease. Valganciclovir, a prodrug of ganciclovir, offers excellent oral bioavailability and is the closest to approval of all the new anti-CMV drugs. High ganciclovir blood levels are achieved without the complications associated with the requirement for long-term iv. access. The monoclonal antibody (mAb) MSL-109, did not offer a significant advantage when added to traditional anti-CMV therapy. Development plans of other agents such as cyclic HPMPC and lobucavir have been put on hold by their respective manufacturers. Adefovir is a nucleotide analogue that possesses anti-CMV activity, but is currently only being pursued for the treatment of hepatitis B virus. Other compounds possessing significant anti-CMV activity, including BAY 38 4766 and GW1263W94 are still in the early stages of development. PMID- 11060673 TI - Recent progress in antiviral chemotherapy for respiratory syncytial virus infections. AB - The recent progress in antiviral chemotherapy against respiratory syncytial virus (RSV) infections was reviewed. RSV infections among high risk individuals, such as premature babies, infants with congenital disease of cardiopulmonary system or immune system and the aged, hospitalised patients with immunosuppressed status are threatened, with high mortality rates and thus need anti-viral chemotherapy. Clinical efficacy of ribavirin and humanized monoclonal antibody (mAb) against RSV infections as well as experimental reports of novel anti-RSV compounds under investigation such as membrane fusion inhibitors were introduced. PMID- 11060674 TI - Novel oral cephalosporins. AB - The therapeutic use of oral cephalosporins to treat infectious diseases continues to challenge clinicians; many attempts have been made over recent years to improve the efficacy and spectrum of these anti-infectives. Many oral cephalosporins are in development and include cefdinir, cefprozil, cefetamet pivoxil, cefcapene pivoxil, cefcanel daloxate hydrochloride in Phase II trials, S 1090 in Phase III trials and the novel compounds E1100, E1101 and BRL-57347. Differences between these drugs are sometimes subtle and the improvement over existing compounds modest, although recent cephalosporins have shown greater activity against Gram-negative bacterial infections. A better knowledge of the pharmacodynamic and pharmacokinetic interrelationships of existing oral cephalosporins is demanded for a more rational use of these compounds and to avoid the subsequent development of resistance. Perhaps with such an approach, the perceived need for new cephalosporins will diminish. PMID- 11060675 TI - beta-Lactamase epidemiology and the utility of established and novel beta lactamase inhibitors. AB - beta-Lactamase inhibitor:beta-lactam combinations remain one of the most successful strategies for the treatment of bacterial infections. Over the last 20 years the number and diversity of serine and metallo active site beta-lactamases has increased dramatically. This review highlights some of the new additions to the beta-lactamase arena and discusses how the commercially available beta lactamase inhibitors are keeping pace with the changing epidemiology of beta lactamases. In addition, we survey the progress with the design of novel inhibitors of serine and metallo-beta-lactamases. Focus is given to the recent advances in the design of metallo-beta-lactamase inhibitors as these enzymes pose a serious emerging threat to the use of all beta-lactam based therapies. PMID- 11060676 TI - Cholic acid derivatives: novel antimicrobials. AB - Mimics of squalamine and polymyxin B (PMB) have been prepared from cholic acid in hope of finding new antimicrobial agents. The squalamine mimics include the polyamine and sulphate functionalities found in the parent antibiotic, however, the positions relative to the steroid nucleus have been exchanged. The PMB mimics include the conservation of functionality among the polymyxin family of antibiotics, the primary amine groups and a hydrophobic chain. Although the squalamine and PMB mimics are morphologically dissimilar, they display similar activities. Both are simple to prepare and demonstrate broad spectrum antimicrobial activity against Gram-negative and Gram-positive organisms. Specific examples may be inactive alone, yet effectively permeabilise the outer membranes of Gram-negative bacteria rendering them sensitive to hydrophobic antibiotics. Problems associated with some of the squalamine and PMB mimics stem from their haemolytic activity and interactions with serum proteins, however, examples exist without these side effects which can sensitise Gram-negative bacteria to hydrophobic antibiotics. PMID- 11060677 TI - Antifungal peptides: potential candidates for the treatment of fungal infections. AB - Many diversely produced natural peptides, as well as those produced semisynthetically and synthetically, have been found to inhibit the growth or even be lethal to a wide range of fungi. Some of these have the potential to aid mankind in combating mycoses caused by emerging pathogens or as a result of the increasing number of antibiotic-resistant fungi. Antifungal peptides may also assist in non-medical fields such as agriculture. For example, introduction by transgenic research of antifungal peptides could improve crop production yields by increasing host resistance to fungal invasion. The aim of this review is to provide information on research on these important peptides. PMID- 11060678 TI - Development of vaccines against human parasitic diseases: tools, current status and perspectives. AB - Vaccines against malaria, leishmaniasis and schistosomiasis are in the most advanced stages of development of all vaccines for human parasitic diseases. Despite the remarkable progress made in identifying and producing protective antigens, at present there are no generally accepted vaccines against parasitic diseases. Vaccines for malaria and leishmaniasis have been taken to clinical trials while vaccines for schistosomiasis are in Phase I/II trials. This review will focus on the most promising antigenic preparations, emphasising the tools, present status and perspectives for development of vaccines against malaria, leishmaniasis and schistosomiasis. PMID- 11060679 TI - Oncolytic viruses as novel anticancer agents: turning one scourge against another. AB - Although the use of viruses as oncolytic agents is an historic concept, the use of genetically modified viruses to selectively target tumour cells is relatively novel and recent. The ability of viruses to efficiently infect and lyse cells, combined with the potential augmentation of this effect by progeny viruses throughout the tumour provide justification for exploitation of these agents in cancer therapy. Before application to humans, though, issues related to tumour cell selectivity, lack of toxicity to normal tissues and the effect of the antiviral immune response, will have to be clarified. The more commonly used oncolytic viruses are based on mutant strains of herpes simplex virus, adenovirus and reovirus. The tumour selectivity of each of these strains is discussed, particularly the complementation of the viral defect by cellular pathways involved in tumourigenesis. The combination of oncolytic viruses with radiation, chemotherapy and gene therapy is also reviewed. Further study of the interaction of viral proteins with cellular pathways involved in cell cycle control will provide the rationale for viral mutants with increased selectivity for tumour cells. PMID- 11060680 TI - Transcription therapy for acute promyelocytic leukaemia. AB - Transcription factors are proteins that regulate gene transcription and expression. In many cases of acute leukaemia chromosomal aberrations are translocations of transcription factors which change their expression and induce the leukaemic phenotype. These abnormal transcription factors are tumour-specific and can be targets for novel treatments approaches. Acute promyelocytic leukaemia (APL) is a distinct and unique subtype of acute myeloid leukaemia (AML) characterised by a reciprocal translocation between chromosomes 15 and 17 t(15q22;17q21). The breakpoints of chromosome 15 and 17 are in the PML and RARalpha genes, respectively, forming the fusion PML-RARalpha gene expressed exclusively and universally in APL. The normal RARalpha is an all-trans retinoic acid- (ATRA-)dependent transcription factor involved in the normal differentiation of myeloid cells. The aberrant fusion PML-RARalpha protein remains sensitive to ATRA and underlies the pathogenesis of the APL. ATRA modulation of gene transcription mediated by PML-RARalpha results in a major clinical response. Almost all newly diagnosed APL cases can be induced into complete remission with ATRA with or without chemotherapy by in vivo differentiation of the APL cells. Randomised clinical trials have shown that the most significant effect of ATRA is an additive or synergistic activity with chemotherapy to improve the long-term outcome of the disease. On the other hand, ATRA with or without induction chemotherapy did not increase the complete remission rate compared to chemotherapy alone. In addition, the relapse rate was significantly lower for patients randomised to induction with concurrent ATRA/chemotherapy than with ATRA followed by chemotherapy. Chemotherapy and/or ATRA maintenance may further improve the long-term outcome compared to no maintenance. PML-RARalpha fusion transcripts can be assayed by RT-PCR to identify PCR positive cells during remission, which are highly predictable of a subsequent haematological relapse. The goal of therapy has been modified to induce a molecular remission with a negative PCR to the PML-RARalpha transcript. This is the first example of an effective response to treatment with a ligand binding to a mutated form of its natural transcription factor. The transcription factor mutation, caused by translocation to another gene, underlies the pathogenesis of the disease. PMID- 11060681 TI - Therapeutic approaches to acquired von Willebrand syndrome. AB - Acquired von Willebrand syndrome (AVWS) is a rare acquired bleeding disorder similar to the congenital von Willebrand disease (VWD) in terms of laboratory findings. Diagnosis of AVWS can be very difficult, with treatment normally taking an empirical form. Although more than 200 cases have been reported since 1968, no retrospective or prospective studies are available on AVWS. Recently, an International Registry on AVWS, gathering data directly from worldwide Departments of Haematology-Oncology and Haemophilia Centres, has been organised by a group working on behalf of the Subcommittee on VWF in the Scientific Standardisation Committee (SSC) of International Society on Thrombosis and Haemostasis (ISTH). Information about an additional 211 AVWS patients is now available, with more detailed data on demography, type of haemorrhage, diagnostic tests for AVWS and management of bleeding episodes. The additional 211 AVWS cases are associated with lymphoproliferative (47%) or myeloproliferative (19%) disorders, cardiovascular diseases, neoplasia (7%) and other miscellaneous diseases (14%). Bleeding episodes of AVWS patients were managed by different compounds including desmopressin (22%), FVIII/VWF concentrates (26%) and high dose immunoglobulin (10%), plasmapheresis (2%), steroids (5%) and immunosuppressive drugs (20%). Based on complied data, we can conclude that none of the therapeutic approaches proposed are 100% effective in all AVWS cases. Therefore, treatment must be customized for each patient according to the underlying disorder, as well as to the type and the severity of bleeding episode and must be targeted to each specific case. PMID- 11060682 TI - Stroke: antithrombin versus antiplatelet therapy. AB - The success of thrombolytic therapy for acute stroke has demonstrated that neurologic outcome can be improved with timely treatment. However, the severely restricted use of thrombolytics has reinforced the need to develop alternative and complementary therapies. Antithrombin and antiplatelet agents represent promising therapeutic approaches for stroke management. Antiplatelet therapy has modestly improved outcome in both acute stroke (aspirin) and in secondary stroke prevention (aspirin with or without dipyridamole; adenosine receptor antagonists), although bleeding and other adverse events associated with antithrombin therapy have largely negated their potential benefit. These findings have prompted innovative solutions to the pharmacokinetic and pharmacodynamic challenges that are crucial to advancing these strategies for acute, primary and secondary stroke therapy. Currently, inhibitors of the platelet surface glycoprotein IIb/IIIa (GP IIb/IIIa, fibrinogen) receptor are being examined in clinical trials while antithrombin therapies focus on thrombin antagonists and inhibitors as well as inhibitors of Factor Xa. Further advances in stroke treatment will include combination therapies. Additionally, the successful design of future drug therapies will result from a more complete understanding of the activity of these agents not only on platelet function and the coagulation cascade, but also for their effects on the endothelium and within the brain parenchyma. The sum of these activities will allow for the maintenance of cerebral blood flow, blood-brain barrier integrity and neuronal function. PMID- 11060683 TI - Pharmacology and clinical experience with amprenavir. AB - Amprenavir (APV, Agenerase) is the fifth protease inhibitor (PI) available for clinical use. It is highly active and its pharmacokinetics allow convenient twice daily administration. It is metabolised by the cytochrome P450 system, leading to a number of drug interactions that have been well defined. Mutations at codons 50 (along with additional changes at codons 46 and 47) lead to the development of resistance, both in vitro and in vivo. In over 200 drug-naive patients, APV-based combination therapy has led to maximal suppression of plasma viral load (generally below 50 copies/ml) in over 50% patients participating in clinical trials lasting 24 - 48 weeks. Benefit has also been demonstrated in over 500 previously treated patients, especially if they are naive to PIs as a therapy. APV-based therapy also appears to be effective in children. Major adverse effects observed in clinical usage have been gastrointestinal (GI) intolerance, skin rash and peri-oral paresthesias. At the present time, it is unclear whether APV offers any advantage over similar types of drugs. Overall, it may be easier to take than some other PIs, but the cost in terms of pill burden is quite high. The toxicity profile (especially the risk of serious skin rashes) may also be an issue. It may be also important in the treatment of isolates that are resistant to other PIs. Its specific role in therapy can only be clarified once the results of ongoing trials (particularly comparative trials of its use in previously treated patients) are available. PMID- 11060684 TI - A review of clinical trials with fluoroquinolones with an emphasis on new agents. AB - This review aims to provide a comparison between the antimicrobial spectra, pharmacokinetics and clinical efficacy of the newer fluoroquinolones with older agents in this class, as well as other antibiotics used to treat lower respiratory and urinary tract infections (LRTIs and UTIs) respectively. Increasing antimicrobial resistance among common uro- and respiratory pathogens has focused attention on the development of fluoroquinolones, which have a broad spectrum of activity and improved tissue penetration. The new and developmental quinolones can be administered on a once-daily basis and exhibit high oral bioavailability, which reduces the need for parenteral therapy in hospitalised patients and may therefore potentially reduce the need for hospitalisation. These attributes, coupled with their expanded spectrum suggest that the newer fluoroquinolones are so far the most ideal agents for the empirical treatment of many common infections. PMID- 11060685 TI - beta-blockers in heart failure: recently completed and ongoing clinical trials. AB - beta-Blockers have emerged as an important therapy in patients with symptomatic left ventricular systolic dysfunction. Early studies demonstrated that beta blocker therapy improved left ventricular function, reduced neurohumoral activity and reduced heart failure symptoms in these patients. While none of these small studies demonstrated a significant benefit in terms of overall survival, several meta-analyses suggested that beta-blocker therapy could, in fact, reduce mortality in patients with left ventricular systolic dysfunction and mild to moderate heart failure symptoms (New York Heart Association class II or III). Three large, recently completed, trials have confirmed the benefit of beta blockade in these patients. This report reviews some of the initial clinical studies of beta-blockade in heart failure, examines the findings of the three large multicentre trials and other relevant research. Finally, ongoing trials designed to assess the relative efficacy of different beta-blockers and evaluate the utility of beta-blockade in specific subsets of patients with heart failure are discussed. PMID- 11060686 TI - Recent developments in the use of vitamin D analogues. AB - The non-classical effects of 1alpha,25-dihydroxyvitamin D(3) (1alpha, 25(OH)(2)D(3)) create possible therapeutic applications for immune modulation (e.g., autoimmune diseases and graft rejection), inhibition of cell proliferation (e.g., psoriasis, cancer) and induction of cell differentiation (e.g., cancer). The major drawback related to the use of 1alpha,25(OH)(2)D(3) is its calcaemic effect, which prevents the application of pharmacological concentrations. Intensive research has led to the development of analogues of 1(2)D(3) characterised by a clear dissociation of the antiproliferative and prodifferentiating capacity from the calcaemic effects. Due to this dissociation, these analogues can be used not only for the treatment of bone disorders but also for non-classical applications. In the present review, a summary is given on the use of the 1alpha,25(OH)(2)D(3) analogues for the treatment of cancer, skin and immune disorders and for the prevention of graft rejection. Moreover a brief overview is given on the use of analogues for secondary hyperparathyroidism. PMID- 11060687 TI - Platelet substitutes and novel platelet products. AB - Despite many advances in the safety, processing and storage of conventional 22 degrees C liquid-stored allogeneic platelet concentrates, there are still significant drawbacks to standard platelet concentrates used in transfusions for patients with thrombocytopenia. Efforts to overcome these shortcomings have been undertaken in both academic and commercial settings, resulting in an array of novel platelet products and substitutes that are currently at various stages of development. This review summarises the recent developments in lyophilised platelets, infusible platelet membranes (IPM), red cells bearing arginine-glycine aspartic acid (RGD) ligands, fibrinogen-coated albumin microcapsules and liposome based agents as putative alternatives to conventional transfusions involving allogeneic platelet concentrates. These various products are designed to replace the use of allogeneic donor platelets with modified or artificial platelets, to augment the function of existing platelets and/or provide a procoagulant material capable of achieving primary haemostasis in patients with thrombocytopenia. Preclinical studies have been encouraging for several of these platelet substitutes and novel platelet products, however, to date, only a few of these products have entered human trials. With the ongoing development of these diverse products, properties necessary for haemostatic effectiveness will become apparent. Safety and efficacy, however, must be demonstrated in preclinical and Phase I - III clinical trials, before these novel agents can be used clinically for patients with thrombocytopenia. PMID- 11060688 TI - DNA vaccines. AB - The premise that DNA coding for antigens produces proteins to stimulate the immune system when inoculated directly into muscle tissues, has the immense attractions of simplicity, versatility and economy. When other vaccination approaches are experiencing practical problems, meeting such challenges as AIDS and malaria, considerable attention has focused on DNA vaccines with entire conferences and a flood of commercial companies devoted to exploring the possibilities. A number of clinical trials for both infectious diseases and cancer have already commenced, even though a number of major issues have to be resolved. PMID- 11060689 TI - Immunologic approaches to antigen discovery for cancer vaccines. AB - Since the early 1990s, scientists have identified an ever-expanding number of antigens to serve as targets for experimental cancer treatments, based on the stimulation of a patient's immune system. Using both immune cells and serum to screen potential candidates, several promising antigens are currently components of vaccines directed against a wide range of tumour types. These antigens vary in their tumour- and tissue-specificity. Their utility as a single reagent or as part of a multi-dimensional approach is as varied as the genes themselves. However, there are already reports indicating that the promise of evoking a clinically beneficial immune response, toward human tumours, is being fulfilled. In this review, we provide a summary of the current status of immunologic approaches to antigen discovery. We also discuss the need for additional, supportive data from non-immunologic techniques, as well as the progression of the preclinical process towards target validation. PMID- 11060690 TI - IL-5: biology and potential therapeutic applications. AB - IL-5 is the predominant cytokine associated with antigen-induced eosinophilic inflammation in the lung. The activation of Th-2 cells leads to the production of IL-5. The pro-eosinophilic effects of IL-5 include: (1) enhanced replication and differentiation of eosinophilic myelocytes; (2) enhanced degranulation of eosinophils; (3) prolonged survival time of eosinophils: and (4) enhanced adhesion of eosinophils. The effects of IL-5 are mediated via the interaction of IL-5 with receptors (IL-5R) that are expressed on the eosinophil cell membrane. Intracellular signalling produced by occupation of the IL-5R by IL-5 occurs via the JAK-STAT system. IL-5 is a 45 kDa glycoprotein consisting of two identical polypeptide chains. The 5'-promoter region of the IL-5 gene contains elements that are down-regulated by glucocorticoids. Anti-IL-5 reagents have the potential to suppress IL-5 activity without the side effects of glucocorticoids. Studies using monoclonal antibodies (mAbs) against IL-5 have established the feasibility of suppressing eosinophilic inflammation by specifically blocking IL-5 activity. Studies with antisense IL-5 are beginning to provide the basis for non glucocorticoid, sequence-specific oligonucleotide inhibitors of IL-5. Research has begun on the development of mAbs and antisense oligonucleotide inhibitors of IL-5 that can be inhaled and applied topically. PMID- 11060691 TI - Soluble cytokine receptors: novel immunotherapeutic agents. AB - Being mediators of immune and inflammatory reactions, abnormal or excessive production of cytokines is often the main cause of the pathology in many types of disease. Targeting cytokines by means of inhibitory drugs may thus offer a valid therapeutic approach in particular diseases. Soluble forms of cytokine receptors (sCR) normally participate in the control of cytokine activity in vivo by inhibiting the ability of cytokines to bind their membrane receptors and from generating a biological response. The ability of sCR to act as cytokine inhibitors, coupled to their specificity, high affinities and low immunogenicities have prompted considerable interest in their use as immunotherapeutic agents. In fact, many types of sCR have been shown to inhibit the biological activity of their cytokines in vitro and in different experimental models. Several sCR, particularly the soluble TNF receptors sTNFR-I (p55) and sTNFR-II (p75), have been modified by linking them to the Fc portion of human immunoglobulin (e.g., 'immunoadhesins') or by the addition of polyethylene-glycol (PEG) (e.g., 'PEGylation'), in order to enhance their affinity and/or biological half-life. These agents have shown significant therapeutic value in clinical trials of patients with rheumatoid arthritis (RA). Indeed, a sTNFR-II:Fc hybrid molecule (etanercept), the first sCR-derived therapeutic agent to receive approval for human use, is already utilised for the treatment of some forms of RA. Additional applications of this drug in other inflammatory conditions are currently being evaluated, while another sCR-derived agent, a human sIL-4R, is undergoing trials for the treatment of asthma. Many other sCR, such as sIL-1R, sIL-5R, sIFNgammaR, may also have significant potential for the treatment of a wide variety of human diseases. PMID- 11060692 TI - Allergen immunotherapy: current and new therapeutic strategies. AB - Allergic individuals respond to an environmental allergen encounter by producing T-cell cytokines, predominantly IL-4 and IL-5, which in turn drive the production of allergen-specific IgE antibodies and recruitment of an eosinophil-rich inflammatory infiltrate. Allergen-specific immunotherapy (SIT) involves the repeated injection of the allergen to specifically downregulate this predominantly Th2-type immune response. SIT is a clinically proven effective treatment for allergic diseases, including rhinoconjunctivitis and asthma. However, despite having been in clinical practice since early this century, its use remains empirical. Best practice protocols are based on clinical experience and include recommendations for selecting patients for treatment, SIT regimes and avoidance of adverse events. More rational and safer SIT regimes will result from new insights into the underlying immune mechanisms for allergic disease, in particular the critical role of helper T-cells in orchestrating this response. The development of recombinant techniques for producing purified allergens and allergen derivatives has led to a dramatic improvement in the ability to standardise allergen preparations and to develop novel vaccines for allergy treatment. Potential vaccines include short peptides based on dominant T-cell epitopes of allergens, allergen fragments and mutant allergens. All of these preparations are designed to target T-cells without binding IgE and inducing local and systemic side effects. Additional strategies under consideration include DNA vaccines and fusion protein constructs incorporating immunomodulatory elements such as bacterial cell proteins, cytokines and immunostimulatory sequences of DNA. Different forms of allergens are being evaluated for the more practical mucosal administration of allergy vaccines. The identification of recombinant allergens suitable for diagnostic use and the development of reliable laboratory assays, based on T-cell function to monitor clinical efficacy of SIT, are important practical outcomes from this research. PMID- 11060693 TI - Novel immunomodulators for topical skin disease therapy. AB - The use of topical corticosteroids has revolutionised the treatment of inflammatory skin diseases. However, problems including pharmacological resistance, as well as the side effect profile of potent topical corticosteroids, has prompted studies to investigate into other topical non-corticosteroidal agents in inflammatory skin diseases. This review outlines the major types of inflammatory skin diseases and discusses emerging therapies based on topical immunosuppressive macrolide antibiotics. In particular, tacrolimus and ascomycin derivatives have been shown to be effective for treating atopic dermatitis with a surprising lack of side effects. It is expected that these agents will play an important role in future dermatological therapy. Accumulating evidence suggests the importance of lipid-derived mediators of inflammation (eicosanoids and platelet-activating factor) in cutaneous inflammatory diseases. The role of these mediators in skin inflammation is also addressed in this review. Though there appears to be a large amount of redundancy in the activities of these lipid mediators, this family of agents could potentially serve as targets for anti inflammatory therapy. Inasmuch as the phospholipase A(2) family of enzymes serve to synthesise both eicosanoids and platelet-activating factor, inhibition at this step could have important therapeutic benefits in designing therapy for inflammatory skin diseases. PMID- 11060694 TI - Adenosine deaminase deficiency as the first target disorder in gene therapy. AB - In the past decade, the advent of gene therapy has been acclaimed as a revolutionary medical intervention, embraced with great enthusiasm. However, recent disappointing results of the considerable clinical trials have also clearly demonstrated that such an initial expectation was an overestimation of gene therapy. There are only a few successful cases despite the 3000 patients who have been treated with various forms of gene therapy. Gene therapy for severe combined immunodeficiency (SCID) caused by adenosine deaminase (ADA) deficiency is one of the few such cases where results have been promising. In particular, peripheral T-lymphocytes-directed gene therapy provides further immunological improvements for patients with ADA-SCID receiving the PEG-ADA treatment whereas gene therapy targeting haematopoietic stem cell has so far proved insufficient for clinical benefits. This report will review crucial problems elucidated in the past five clinical trials for ADA-SCID and gives an outline of the next generation of stem cell gene therapy in Japan. PMID- 11060695 TI - Therapeutic potential of adenosine kinase inhibitors. AB - Adenosine kinase (AK; EC 2.7.1.20) is a key intracellular enzyme regulating intra and extracellular concentrations of adenosine (ADO), an endogenous modulator of intercellular signalling that reduces cell excitability during tissue stress and trauma. The inhibitory effects of ADO are mediated by interactions with specific cell-surface G-protein coupled receptors (GPCR), which regulate membrane cation flux, membrane polarisation and the release of excitatory neurotransmitters. Inhibition of AK potentiates local extracellular ADO levels at cell and tissue sites which are undergoing accelerated ADO release. Thus, AK inhibition represents a mechanism to selectively enhance the endogenous protective actions of ADO during cellular stress while potentially minimising the non-specific effects associated with the systemic administration of ADO receptor agonists. Novel, potent AK inhibitors have recently been synthesised that demonstrate high specificity for this particular enzyme as compared to other ADO metabolic enzymes, transporters and receptors. AK inhibitors have been shown to increase ADO concentrations in various systems in vitro, as well as in an in vivo model of neurotoxicity. In addition, AK inhibitors have demonstrated efficacy in animal models of epilepsy, cerebral ischaemia as well as pain and inflammation, thus suggesting their potential therapeutic utility for these conditions. PMID- 11060696 TI - Therapeutic advances in small cell lung cancer. AB - Small cell lung cancer (SCLC) is characterised by neuroendocrine differentiation, early metastatic potential and initial responsiveness to cytotoxic therapy. Unfortunately, despite recent therapeutic advances, most patients relapse and the overall five-year survival rate is only 5%. Standard treatment of SCLC consists of platinum-based combination chemotherapy, with thoracic irradiation added for patients with limited-stage disease. Several newer chemotherapeutic drugs have recently been shown to have significant activity in patients with untreated or relapsed SCLC. These agents include: the topoisomerase I inhibitors, topotecan and irinotecan; the taxanes, paclitaxel and docetaxel; the pyrimidine analogue, gemcitabine; and the vinca alkaloid, vinorelbine. Recent advances in our understanding of the molecular events involved in the pathogenesis and progression of SCLC have led to the identification of a variety of potential targets for novel therapeutic interventions. Strategies aimed at inhibiting the myriad of growth factor pathways that control the proliferation of SCLC cells, include: broad spectrum neuropeptide antagonists (e.g., substance P analogues); growth factor/receptor-specific inhibitors (e.g., anti-GRP monoclonal antibodies, bradykinin antagonist dimers); and a variety of selective protein kinase inhibitors. The importance of cell death pathways in carcinogenesis and treatment resistance has led to several novel strategies targeting apoptotic mediators, such as bcl-2, that are frequently dysregulated in SCLC (e.g., bcl-2 antisense). Our current challenges are to further refine these promising therapeutic strategies, efficiently evaluate their activity in the clinical setting and integrate them into more effective treatment regimens to improve the overall prognosis of patients with SCLC. PMID- 11060697 TI - Low molecular weight heparin in unstable coronary artery disease. AB - Since the recognition that any thrombosis overlying a ruptured atherosclerotic plaque is a central component of the pathogenesis of unstable coronary artery disease (CAD), a number of antithrombotic treatment strategies have been investigated in randomised clinical trials. Aspirin reduces the occurrence of symptomatic and silent ischaemia, myocardial infarction (MI) and death in patients with unstable CAD, both in the acute phase and during continued long term treatment and is now considered routine therapy. The addition of unfractionated heparin infusion further reduces cardiac events during the treatment period, but is not associated with any sustained benefits during long term follow-up. Low molecular weight heparins (LMWHs) are completely absorbed by the sc. route. A predictable anticoagulant effect is maintained by sc. injections every 12 - 24 h without laboratory monitoring. The FRISC trial demonstrated that, in conjunction with aspirin, the LMWH dalteparin reduced death and MI by more than 50% in the acute phase. Four similar trials have directly compared LMWH with unfractionated heparin and demonstrated at least the same efficacy in the acute phase, treated between three and eight days. The LMWH enoxaparin was more effective at reducing death or MI than unfractionated heparin infusion during the three days of treatment, an effect which lasted up to 12 months. Prolonged out patient treatment beyond the acute phase has been evaluated in four trials. It was demonstrated, in two of these trials, that continuing twice-daily injections of dalteparin further reduced the risk of death, MI and need for revascularisation at least during the initial six weeks of treatment. This effect was confined, however, to patients with signs of myocardial damage, i.e., elevation of troponin, at admission. During prolonged out-patient treatment, there is an increased risk of severe bleeding due to the combination of LMWH with aspirin. Based on successful results, the LMWHs, having the convenience of treatment and the option for continuation in an out-patient setting, should replace unfractionated heparin as the routine treatment in unstable CAD. If an early invasive procedure is considered, the LMWH therapy should be continued until the intervention as a 'bridge-to-revascularisation'. New antiplatelet agents are also emerging as additional useful tools for treating these patients, thus it is urgent to evaluate the comibination of these new therapies with the LMWHs. PMID- 11060698 TI - Novel triazole antifungal agents. AB - The risk of opportunistic infections is greatly increased in patients who are immunocompromised due to AIDS, cancer chemotherapy and organ or bone marrow transplantation. Candida albicans is often associated with serious systemic fungal infections, however other Candida species such as Candida krusei, Candida tropicalis and Candida glabrata, as well as Cryptococcus neoformans and filamentous fungi such as Aspergillus, have also emerged as clinically significant fungal pathogens. Two triazole antifungal agents, fluconazole and itraconazole, were introduced over a decade ago and since then have been used extensively for the prophylaxis and treatment of a variety of fungal infections. Although both drugs are effective and have their place in therapy, limitations regarding the utility of these agents do exist. For example, fluconazole is not effective for the prophylaxis or treatment of Aspergillus species and has limited activity against C. krusei and C. glabrata. The use of itraconazole has been limited secondary to concerns regarding unpredictable bioavailability. The rising incidence of fungal infections and the reported increase of non-albicans candidal infections noted over the past two decades highlight the need for new antifungal agents with improved spectra of activity. Several new triazole agents are in various phases of preclinical and clinical trials and may be available for human use in the near future. Three such agents voriconazole, posaconazole and ravuconazole are reviewed and compared with existing agents. PMID- 11060699 TI - Ebselen: prospective therapy for cerebral ischaemia. AB - Stroke occurs due to haemorrhage or occlusive injury and results in ischaemia and reperfusion injury. A variety of destructive mechanisms are involved including oxygen radical generation, calcium overload, cytotoxicity and apoptosis as well as the generation of inflammatory mediators. Ebselen, 2-phenyl-1, 2 benzisoselenazol-3(2H)-one (PZ 51, DR3305), is a mimic of GSH peroxidase which also reacts with peroxynitrite and can inhibit enzymes such as lipoxygenases, NO synthases, NADPH oxidase, protein kinase C and H(+)/K(+)-ATPase. Ebselen is in a late stage of development for the treatment of stroke. The molecular actions of ebselen contribute to its anti-inflammatory and anti-oxidant properties, which have been demonstrated in a variety of in vivo models. Numerous in vitro experiments using isolated LDL, liposomes, microsomes, isolated cells and organs have established that ebselen protects against oxidative challenge. Unlike many inorganic and aliphatic selenium compounds, ebselen has low toxicity as metabolism of the compound does not liberate the selenium moiety, which remains within the ring structure. Subsequent metabolism involves methylation, glucuronidation and hydroxylation. Experimental studies in rats and dogs have revealed that ebselen is able to inhibit both vasospasm and tissue damage in stroke models, which correlates with its inhibitory effects on oxidative processes. Results from randomised, placebo-controlled, double-blind clinical studies on the neurological consequences of acute ischaemic stroke, subarachnoid haemorrhage and acute middle cerebral artery occlusion, have revealed that ebselen significantly enhances outcome in patients who have experienced occlusive cerebral ischaemia of limited duration. The benefit achieved with ebselen is closely related to the rapidity with which the treatment is initiated, following the onset of the stroke attack. Safety and tolerability are good and no adverse effects have become apparent. Ebselen is currently at the pre-registration stage for subarachnoid haemorrhage and stroke in Japan. PMID- 11060700 TI - Therapeutic developments in multiple sclerosis. AB - Recently there have been considerable advances made in the treatment of multiple sclerosis. For the first time since its initial clinical description in the 1800s, there are now available several medications which unequivocally exert favourable clinical effects through the lowering of the biological activity of the human illness. The therapeutic efficacy of IFN-beta preparations seems particularly well established in this regard on the basis of five large, independent, trials of this agent. These trials have demonstrated remarkably consistent reductions in both attack rates and disability levels using a combination of clinical and magnetic resonance imaging outcome measures. The therapeutic benefit of glatiramer acetate also has been well established, although there is less available data on this agent than there is for interferon. It is important to recognise, however, that, although these agents represent an important first step in the management of patients with multiple sclerosis, they are only partial therapies. In order to actually cure the illness or even to substantially improve patient outcome we need considerably better agents than we have currently. Nevertheless, it is likely that, with improved knowledge of the role that interferon beta plays in the pathogenesis of multiple sclerosis and with better understanding of the mechanism by which glatiramer acetate exerts its therapeutic effect, greatly improved therapeutic agents will be available in the future. In addition, it seems likely that, in the future (by analogy to the experience in oncology), we will begin utilising combinations of therapies in order to better control the biological activity of this debilitating disease. Such combination therapy will almost certainly include combinations of partially effective agents as well as combinations of these agents with other medications (e.g., the immunosuppressive drugs) which, by themselves, have only been demonstrated to exert marginal clinical benefits on the course of illness. Moreover, it also seems likely that, increasingly, therapeutic strategies that enhance or promote myelin repair will become a major focus of clinical research in this area. PMID- 11060701 TI - Anti-inflammatory drugs: a hope for Alzheimer's disease? AB - Human brain cells are capable of initiating and amplifying a brain specific inflammatory response involving the synthesis of cytokines, acute-phase proteins, complement proteins, prostaglandins and oxygen radicals. In Alzheimer's disease (AD), all signs of an inflammatory microglial and astroglial activation are present inside and outside amyloid depositions and along axons of neurones with neurofibrillary tangles. Cell culture and animal models suggest a bidirectional relationship between inflammatory activation of glial cells and the deposition of amyloid. Although it remains unclear which of the different pathophysiological processes in AD may be the driving force in an individual case, the inflammatory activation may increase the speed of cognitive decline. Epidemiological studies point to a reduced risk of AD among users of anti-inflammatory drugs. Therefore, anti-inflammatory drugs have become the focus of several new treatment strategies. A clinical trial with the non-steroidal anti-inflammatory drug (NSAID) indomethacin showed promising results, while a clinical trial with steroids did not show a beneficial effect. Further trials with NSAIDs such as unselective cyclooxygenase (COX) and selective cyclooxygenase-2 (COX-2) inhibitors are on their way. COX inhibitors may not only act on microglial and astroglial cells but also reduce neuronal prostaglandin production. New data suggest that prostaglandins enhance neurotoxicity or induce pro-inflammatory cytokine synthesis in astroglial cells. Amongst these promising new strategies to reduce microglial or monocyte activation, interfering with intracellular pathways has been shown to be effective in various cell culture and animal models but clinical studies have not yet been performed. PMID- 11060702 TI - Oestrogen and nerve growth factor - neuroprotection and repair in Alzheimer's disease. AB - The neurogenetics and neuropathology of Alzheimer's disease (AD) are still largely unknown, even though recent work has clarified some genetic components in this common and devastating neurodegenerative disease. Most of the genetic mutations have been shown to be, at least in the early onset type of AD, related to the function of a large transmembrane protein, amyloid precursor protein (APP). This protein is cleaved into various smaller fragments that are either soluble or aggregating. It is thought that this processing of APP is inherently important for the initiation and progression of AD. Recent animal models have suggested that it is not the formation of beta-amyloid plaques per se, but the altered processing of APP and the subsequent loss of soluble APP, that sets the stage for the massive neuronal cell loss which occurs in AD. We would like to propose a three-way relationship between oestrogen, APP and nerve growth factor (NGF) in the neural pathways of the brain which are involved in learning and memory - the limbic system. The degeneration of the cholinergic innervation from the basal forebrain to the hippocampal formation in the temporal lobe is thought to be one of the factors determining the progression of memory decay, both during normal ageing and AD. Oestrogen and NGF are among the neuroprotective agents that have shown some potential for the treatment of AD. Previous results of treatment with these two agents and their relationship to the amyloid proteins, will be discussed in this review. PMID- 11060703 TI - Neuroprotection in cerebrovascular disease. AB - The role of neuroprotection in the management of acute cerebrovascular disease is reviewed. Neuroprotection is a valuable adjunct to thrombolytic therapy in acute cerebral ischaemia. Various pharmacological approaches for neuroprotection are based on the current knowledge of molecular events in the pathophysiology of cerebral ischaemia. Reperfusion injury following restitution of circulation is also considered to be mediated by free radicals. Various strategies include free radical scavengers, anti-excitotoxic agents, apoptosis (programmed cell death) inhibitors, anti-inflammatory agents, metal ion chelators, ion channel modulatory, antisense oligonucleotides and gene therapy. The various agents aim to prevent the progression of ischaemic cascade therefore reducing brain damage and some of these intervene at more than one point in the ischaemic cascade. Neuroprotection is considered as an adjunct to therapies designed to improve cerebral circulation such as thrombolytic agents for arterial thrombosis. Clinical effectiveness of some of the strategies has not be proven in clinical trials, some of which had to be abandoned due to adverse effects outweighing the beneficial effects. Efforts to develop new neuroprotective agents continue and prospects for the introduction of an effective neuroprotective agent(s) in the next few years are good. Apart from acute cerebrovascular disease, neuroprotective therapy has a role in preventing cerebral ischaemia in high risk cardiovascular procedures as well as in neurodegenerative disorders which has some common pathomechanisms with cerebrovascular disease. Currently, the most promising agents are free radical scavengers. In the near future, gene therapy approaches are likely to prove more effective in neuroprotection. PMID- 11060704 TI - Current status of viral gene therapy for brain tumours. AB - Malignant glial tumours represent the majority of primary brain tumours. Despite the use of many adjunctive treatment strategies in addition to surgery, the prospect of cure or even long-term survival is poor. In the last decade, there has been an explosion of interest in the development of delivery systems that will allow the expression of exogenous genes in the CNS. For the most part, these systems are based upon modified viruses. To date, the greatest experience has been with retroviruses, herpes simplex virus 1 (HSV), adenovirus and adeno associated virus (AAV). This review will outline the biology of these viral vectors, modifications permitting in vivo administration and their respective advantages and disadvantages for the treatment of malignant brain tumours. The present obstacles to gene therapy strategies will also be described. To date, no convincing clinical trial has emerged that provides objective proof of the superiority of gene therapy strategies as compared to conventional treatment. PMID- 11060705 TI - Immunotherapeutic approaches to paraneoplastic neurological disorders. AB - Paraneoplastic neurological disorders (PNDs) are disturbances of the nervous system function in cancer patients, which are not due to a local effect of the tumour or its metastases. This review focuses on the neuromuscular PND and central nervous system (CNS) PND, which are by far the most problematic group to treat. Most of these clinically well-defined syndromes are associated with lung cancer, especially small cell lung cancer (SCLC), lymphoma and gynaecological tumours. Since auto-antibodies directed against proteins expressed in neurones and tumour cells have been found, PNDs are suspected to be autoimmune. In neuromuscular PND, immunosuppressive therapies, plasmapheresis and iv. immunoglobulins (iv. Ig) have proven to be effective. In PND of the CNS, therapy of the tumour itself or immunosuppressive drugs seem to have little effect on the neurological syndromes. Plasmapheresis reduces the auto-antibody titre in the sera of these patients, but does not lead to a clinical improvement. This review presents an overview on the pathogenesis of PND and the possible benefit of an early immunosuppressive or immunomodulatory treatment, especially treatment with iv. immunoglobulins. PMID- 11060706 TI - Neurokinin receptor antagonists. AB - Studies on tachykinin peptides and the corresponding neurokinin receptors (NKr) have increased dramatically recently due to the discovery of selective, orally active, metabolically stable and sometimes CNS penetrating NKr antagonists. After demonstrating the potential use for NKr antagonists in animal models, some compounds have recently progressed into clinical trials and a few results have been published. NKr antagonists have demonstrated efficacy for the treatment of emesis and depression, while results in other areas have been disappointing. Nonetheless, this area is coming to the exciting time of proof of concept in humans. Demonstration of the involvement of tachykinin peptides in biological functions continues to grow, as do the potential indications for NKr antagonists. More drug candidates are undergoing clinical trials for various conditions and these results could widen the potential use for NKr antagonists. PMID- 11060707 TI - Apoptosis modulators in the therapy of neurodegenerative diseases. AB - Apoptosis is a prerequisite to model the developing nervous system. However, an increased rate of cell death in the adult nervous system underlies neurodegenerative disease and is a hallmark of multiple sclerosis (MS) Alzheimer's- (AD), Parkinson- (PD), or Huntington's disease (HD). Cell surface receptors (e.g., CD95/APO-1/Fas; TNF receptor) and their ligands (CD95-L; TNF) as well as evolutionarily conserved mechanisms involving proteases, mitochondrial factors (e.g. , Bcl-2-related proteins, reactive oxygen species, mitochondrial membrane potential, opening of the permeability transition pore) or p53 participate in the modulation and execution of cell death. Effectors comprise oxidative stress, inflammatory processes, calcium toxicity and survival factor deficiency. Therapeutic agents are being developed to interfere with these events, thus conferring the potential to be neuroprotective. In this context, drugs with anti-oxidative properties, e.g., flupirtine, N-acetylcysteine, idebenone, melatonin, but also novel dopamine agonists (ropinirole and pramipexole) have been shown to protect neuronal cells from apoptosis and thus have been suggested for treating neurodegenerative disorders like AD or PD. Other agents like non-steroidal anti-inflammatory drugs (NSAIDs) partly inhibit cyclooxygenase (COX) expression, as well as having a positive influence on the clinical expression of AD. Distinct cytokines, growth factors and related drug candidates, e.g., nerve growth factor (NGF), or members of the transforming growth factor-beta (TGF-beta ) superfamily, like growth and differentiation factor 5 (GDF-5), are shown to protect tyrosine hydroxylase or dopaminergic neurones from apoptosis. Furthermore, peptidergic cerebrolysin has been found to support the survival of neurones in vitro and in vivo. Treatment with protease inhibitors are suggested as potential targets to prevent DNA fragmentation in dopaminergic neurones of PD patients. Finally, CRIB (cellular replacement by immunoisolatory biocapsule) is an auspicious gene therapeutical approach for human NGF secretion, which has been shown to protect cholinergic neurones from cell death when implanted in the brain. This review summarises and evaluates novel aspects of anti-apoptotic concepts and pharmacological intervention including gene therapeutical approaches currently being proposed or utilised to treat neurodegenerative diseases. PMID- 11060708 TI - Therapeutic potential of positive AMPA receptor modulators in the treatment of neurological disease. AB - Excitatory neurotransmission in the CNS depends heavily upon alpha-amino-3 hydroxy-5-methylisoxazole-4-propionate (AMPA)-type glutamate receptors. Derangements in AMPA receptor mediated synaptic transmission may be a contributing factor in neurological and neurodegenerative diseases and could be a target for therapeutic intervention. Recently, drugs that positively modulate AMPA receptors have been identified, having differential effects upon certain AMPA receptor subunits and different effects upon physiological properties of AMPA receptors. These drugs facilitate AMPA receptor mediated processes and may have beneficial therapeutic effects. For example, certain AMPA modulators facilitate long-term potentiation, which is considered a cellular mechanism that may be important for memory storage and they also facilitate memory encoding in behavioural experiments. Thus, AMPA modulators might ameliorate memory deficits that occur in dementia, such as Alzheimer's disease (AD). However, AMPA receptor mediated excitotoxicity may occur with excessive AMPA receptor activation which occurs in seizures or ischaemia and positive AMPA modulators could promote neuronal injury in those conditions. Ultimately, the clinical utility of positive AMPA modulators will be dependent upon understanding the role of AMPA receptors in certain neurological disorders, identifying receptor subtypes involved in specific neurological disorders and developing drugs with selective actions upon specific AMPA receptor properties that also possess receptor subtype specificity. Currently available drugs have provided significant insight into the physiology and structural determinants of important AMPA receptor properties and some insight into potential clinical uses as well as potential dangers of such drugs. PMID- 11060709 TI - Mechanisms of action and resistance to tubulin-binding agents. AB - Tubulin binding agents constitute an important class of antimitotics and are widely used for the treatment of solid tumours an haematopoietic malignancies. These compounds, currently represented by the vinca alkaloids and the taxanes, differ from most of the other clinically useful antimitotics in that their target is not nucleic acids, but the mitotic spindle, which is an essential component of the mitotic machinery. Recent data on the mechanisms of action of and mechanisms of resistance to tubulin binding agents are presented. The importance of microtubule dynamics is emphasised, in particular in relationship to the usefulness of drug combinations. Concerning the reported resistance mechanisms, an emerging body of data show that altered microtubule structure may be involved in reduced sensitivity to these compounds. Promising new molecules, including those derived from marine organisms are described. PMID- 11060710 TI - Immune-adhesion molecules in the prevention of allograft rejection and reperfusion injury. AB - Control of the immune system is of indispensable importance for graft acceptance and function. Immunological changes in the graft before and after organ harvesting, the transplantation procedure itself and the organ recipients clinical state contribute to the immune response. Leukocyte trafficking [1] into a graft is regulated by various signal transducing molecules, which have been characterised during the past years. Ligand molecules on endothelial cells and in the organ parenchyma are the counterparts for leukocyte adhesion and tissue infiltration. The expression of these ligand molecules is regulated by soluble factors and cell-cell interactions [2]. The regulation of tissue inflammation and repair mechanisms involving components of the immune system therefore depends on a number of cell-surface interactions. The processes of intravascular adhesion, transmigration and infiltration by leukocytes and platelets are mainly mediated by receptor ligand interactions with target cells (cell-cell) and extracellular matrix proteins (cell-matrix). The main molecular families of adhesion receptor/ligand molecules have been identified. Today, we are still far from understanding this network of interactions. The numbers of molecules and factors involved are still increasing. This review summarises the currently available knowledge on the intervention in this system by monoclonal antibodies (mAbs), peptides and blocking agents. From this review, it is evident that further investigations are justified. PMID- 11060711 TI - Influence of photodynamic therapy on immunological aspects of disease - an update. AB - Photodynamic therapy (PDT) utilises light-absorbing compounds combined with directed photo-irradiation to produce clinical effects. This review updates advances in the understanding of the biochemical pathways triggered by PDT within cells, its influence upon different immune parameters and progress in the use of PDT against human immune-mediated disease. Several works have further defined the notable capacity of PDT to foster anticancer immunity. PMID- 11060712 TI - Ziprasidone: comprehensive overview and clinical use of a novel antipsychotic. AB - Ziprasidone (5-[2-(4-(1,2,-benzisothiazol-3-yl) piperazin-l-yl] ethyl]-6 -chloro indolin-2-one hydrochloride hydrate) is a novel antipsychotic with a pattern of receptor occupancy and preclinical attributes predictive of broad therapeutic efficacy and a favourable tolerability profile in the treatment of psychotic illness. Clinical trials indicate that ziprasidone is effective against positive, negative and affective symptoms in schizophrenia and schizoaffective disorder with minimal motor, cognitive, weight gain, prolactin related, or anticholinergic side effects. In addition, an im. formulation appears to be rapidly effective with significantly less motor side effect liability than haloperidol. PMID- 11060713 TI - An assessment of rufinamide as an anti-epileptic in comparison with other drugs in clinical development. AB - This article evaluates rufinamide, a new anti-epileptic drug (AED) in Phase III development. This review is done against the background of therapeutic challenges of epilepsy, old established AEDs, newly introduced AEDs and AEDs in clinical development. Pharmacological properties of 12 AEDs in clinical trials (Phases I - III) are compared: ADCI, AWD 131-138, DP-VPA, ganaxolone, levetiracetam, losigamone, pregabalin, remacemide hydrochloride, retigabine, rufinamide, soretolide and TV1901. One of these, levetiracetam has been approved in the USA and is waiting approval in other countries. The protective index of rufinamide, as shown in rodent models of epilepsy, is much higher than that of most common AEDs. Features which make it a desirable AED are: (i) a broad spectrum of anti epileptic actions including both partial and symptomatic generalised epilepsy; (ii) a statistically significant reduction in seizure frequency in clinical trials; (iii) efficacy and safety shown in a broad range of age groups including children and the elderly; (iv) rapid oral absorption enabling quick titration to effective dose and (v) a benign adverse event profile. Most of the events did not lead to discontinuation in clinical trials. These features offer considerable advantages over the existing anti-epileptic drugs. It is one of the two drugs in development which have reached Phase III and is expected to be approved by the year 2001 - 2002. PMID- 11060714 TI - Zaleplon - a review of a novel sedative hypnotic used in the treatment of insomnia. AB - Zaleplon (N-[3-(3-cyanopyrazolo[1,5-a] pyrimidin-7-yl) phenyl]-N-ethyl acetamide) is a non-benzodiazepine recently introduced for clinical use. This agent is indicated for the short-term treatment of insomnia. Preclinical studies have shown that the benzodiazepines triazolam and Ro17-1812 can substitute for zaleplon in animals trained to distinguish zaleplon from saline. The benzodiazepine antagonist flumazenil can antagonise the discriminative stimulus effect of zaleplon. These findings suggest that zaleplon is recognised by animals as a benzodiazepine agent. Zaleplon is active after ip. and oral administration in a variety of motor performance tests, including locomotor activity, rotarod and the loaded grid. Zaleplon has been shown to be active in a number of different anticonvulsant models, including the pentylenetetrazole, isoniazid and electroshock models. The compound is also reported to be active against convulsions induced by bicuculline, picrotoxin and strychnine. Studies in anxiolytic models suggest that zaleplon may have weak anxiolytic activity. From preclinical studies, it appears zaleplon possesses a reduced risk of tolerance compared to triazolam, is less likely to potentiate the effects of ethanol and is unlikely to produce amnestic effects. In man, zaleplon is rapidly absorbed and undergoes extensive presystemic metabolism. The compound has a plasma half-life of approximately one hour and is metabolised primarily via the aldehyde oxidase system to form 5-oxo-zaleplon. This metabolite, along with other minor metabolites formed in vivo, do not appear to contribute to the activity of zaleplon. Metabolites of zaleplon are excreted primarily via the urine. Phase I studies suggest that single daytime doses of zaleplon up to 15 mg are well tolerated. Short-term impairment of performance occurs when zaleplon is administered during the day at doses epsilon 20 mg. However, given the short half life of the compound, significant impairment of daytime performance is unlikely if zaleplon is administered at bedtime or shortly after retiring for the evening. Results from Phase II/III studies suggest that zaleplon (5 - 20 mg) produces a dose-dependent reduction in sleep latency in patients suffering from primary insomnia. The clinical efficacy of zaleplon persists for at least four weeks at doses of 10 mg and 20 mg. Studies in patients with a history of drug abuse suggest that the abuse potential of zaleplon (at doses above the therapeutic dose range) is similar to that seen with the benzodiazepine triazolam. PMID- 11060715 TI - Nicaraven for the treatment of cerebral vasospasm in subarachnoid haemorrhage. AB - Cerebral vasospasm is a complication of subarachnoid haemorrhage and can cause cerebral ischaemia. Antivasospastic agents are used to relieve vasospasm after subarachnoid haemorrhage. A large number of agents with varying modes of action currently being investigated are reviewed. Pharmacology and clinical trials of nicaraven are discussed. The drug has been found to have both antivasospastic as well as neuroprotective effects. Clinically, the most documented efficacy of nicaraven is in the management of vasospasm associated with subarachnoid haemorrhage based on its free radical scavenging effect. Other potential areas for application are cerebral oedema associated with intracerebral haemorrhage and for neuroprotection in cerebral infarction. Nicaraven is in pre-registration by Chugai Pharma Ltd. in Japan for the treatment of vasospasm following subarachnoid haemorrhage. The regulatory atmosphere in Japan regarding the approval of neuroprotectives is reviewed and nicaraven is likely to be approved by the year 2001 when the patent on it expires. PMID- 11060716 TI - Remacemide: current status and clinical applications. AB - Remacemide (RMC) is a non-competitive, low-affinity N-methyl-D-aspartate (NMDA) receptor antagonist that does not cause the behavioural and neuropathological side effects seen with other NMDA receptor antagonists. RMC and its active metabolite, AR-R 12495 AR, which has moderate affinity for the NMDA receptor, also interact with voltage-dependent neuronal sodium channels. Both agents show efficacy in a variety of animal models of epilepsy, parkinsonism and cerebral ischaemia. There is no evidence for teratogenicity or genotoxicity. RMC delays the absorption of L-dopa and elevates the concentrations of drugs metabolised by the hepatic cytochrome P450 3A4 isoform. RMC and AR-R 12495 AR have moderate protein binding and linear pharmacokinetics. Controlled studies show evidence of efficacy in treating epilepsy and Parkinson's disease. Post-surgical outcomes in RMC-treated patients at risk for intra-operative cerebral ischaemia are also encouraging. Adverse effects are related to the gastrointestinal and central nervous systems. RMC is a promising drug with numerous potential applications for both acute or chronic conditions associated with glutamate-mediated neurotoxicity. PMID- 11060717 TI - Pharmacology and clinical experience with repaglinide. AB - Repaglinide (NovoNorm((R))) is a novel oral antidiabetic agent, the first of a new class of insulin secretagogues known as the prandial glucose regulators to be approved for use in patients with Type 2 diabetes. Prandial glucose regulation is aimed at restoring the first-phase insulin response that follows consumption of a meal, which is missing in patients with Type 2 diabetes. After repaglinide administration, the resulting insulin profile reflects that of healthy individuals more closely, providing tighter glycaemic control and reducing the risk of hypoglycaemic events. Repaglinide is quickly absorbed and rapidly eliminated through biliary excretion, making it suitable for use in patients with renal impairment. It appears in the bloodstream within 15 to 30 min of dosing, stimulating short-term insulin release from the pancreatic beta-cells by binding to a unique site on the beta-cell membrane. Rapid elimination ensures that postprandial insulin levels quickly return to preprandial levels as the high prandial glucose level subsides. Repaglinide is given on a 'one meal, one tablet; no meal, no tablet' basis. It is particularly effective in patients who have not previously been treated with an oral antidiabetic agent, significantly reducing glycosylated haemoglobin (HbA(1c)) levels by 1.6%. It also offers increased mealtime flexibility and safety, compared with other oral antidiabetic agents. As a result of the short plasma half-life and lack of accumulation of repaglinide with repeated dosing, the risk of between-meal and nocturnal hypoglycaemia is substantially reduced compared with other oral antidiabetic agents. Repaglinide acts synergistically with metformin, consistently improving glycaemic control in patients who were insufficiently controlled by metformin alone. Results from recent studies have shown similar synergistic effects with neutral protamine Hagedorn (NPH)-insulin or troglitazone. PMID- 11060718 TI - Ongoing trials in Alzheimer's disease. AB - Researchers have sought to understand the underlying pathophysiology of Alzheimer's disease (AD) ever since Dr A Alzheimer first described the condition in 1907. Unfortunately however, until recently, they have done so with limited success. This lack of clarity has deterred advancements in therapeutic drug research beyond all but the purely symptomatic treatment relief currently available. However, through spatio-temporal analysis of the two types of cerebral lesions that characterise the disorder (senile plaques and neurofibrillary tangles) and the compilation of genetic data concerning familial AD, there now exists the foundation for a more comprehensive understanding of the disease. Although symptomatic cholinergic strategies have beneficial effects, their benefits are modest and current research has turned to the development of other promising strategies, including oestrogen replacement, anti-inflammatory agents, free radical scavengers, anti-oxidants and monoamine oxidase-B (MAO-B) inhibitors. Many of these strategies may have some merit, however further analysis and structured research are necessary before a definitive decision can be made about their efficacy and possible role in AD therapy. Strategies that are directed at halting the underlying biochemical changes in AD are nearing clinical testing and offer the promise for meaningful therapeutic outcomes. PMID- 11060719 TI - Ongoing trials in HIV protease inhibitors. AB - The development of antiretrovirals has led to a revolution in the care of patients infected with HIV. What was once a uniformly fatal syndrome has become a more treatable, chronic, infectious disease. Central to this revolution have been the protease inhibitors, a class of drugs with potent antiretroviral activity. The first member of this class was approved for use in 1995 and there are now five protease inhibitors approved by the US Food and Drug Administration (FDA): amprenavir, indinavir, nelfinavir, ritonavir and saquinavir. As a result of the magnitude of the HIV pandemic coupled with the clinically proven efficacy of protease inhibitors, there are currently hundreds of ongoing clinical trials with these agents. Trial designs include comparisons between the various licensed protease inhibitors, comparisons of protease inhibitors to other classes of potent antiretroviral drugs, investigations with new protease inhibitors, investigations of protease inhibitor-related toxicities and attempts at simplifying current dosing regimens. PMID- 11060720 TI - Therapeutic potential of cyclic nucleotide phosphodiesterase inhibitors in heart failure. AB - There are several reasons to believe that agents that augment cAMP-mediated signalling in cardiac myocytes should have beneficial effects in patients with heart failure. However, clinical trials of first-generation cyclic nucleotide phosphodiesterase (PDE3) inhibitors, which raise cAMP content by blocking its hydrolysis, have shown that chronic administration of these drugs affect survival adversely. The problem may be the non-selective activation of a broad spectrum of cAMP-regulated cellular responses these agents elicit. More selective (or alternatively selective) cyclic nucleotide PDE inhibitors might improve results by evoking a more restricted set of cellular responses. PMID- 11060721 TI - Therapeutic potential of complement inhibitors in myocardial ischaemia. AB - Under normal conditions, the complement system functions to eradicate microbes and other membrane bound pathogens. In other situations, complement activation comprises a pivotal mechanism for mediating tissue demolition in inflammatory disorders, including ischaemia/reperfusion injury. Complement-mediated tissue damage has long been recognised as a significant contributor to myocardial reperfusion injury. However, clinical use of complement inhibitors to reduce the extent of irreversible tissue injury related to reperfusion, remains in the early stages of development. Activation of the complement system generates anaphylatoxins, opsonins and the lytic moiety known as the membrane attack complex (MAC). In addition, fragments of the complement cascade proteins (e.g., C3a and C5a) secondarily initiate processes deleterious to myocytes by recruiting and stimulating inflammatory cells, such as neutrophils and macrophages, within the area of reperfusion. Damaged tissue itself, is capable of upregulating the genes that encode the formation of complement proteins leading to assembly of the MAC, which in turn further advances tissue injury. All of these factors contribute to the development of myocardial infarction subsequent to ischaemia and reperfusion. This paper provides an overview of how the complement system operates and examines the various inhibitors, both endogenous and exogenous, that regulate the complement cascade. Activation and inhibition of the complement system will be discussed primarily in the context of myocardial ischaemia and reperfusion injury. PMID- 11060722 TI - Therapeutic potential of matrix metalloproteinase inhibitors in atherosclerosis. AB - The activity of matrix-degrading metalloproteinases (MMPs) is essential for many of the processes involved in atherosclerotic plaque formation, for example, infiltration of inflammatory cells, smooth muscle cell migration and proliferation and angiogenesis. Furthermore, matrix degradation by MMPs may cause the plaque instability and rupture that leads to the clinical symptoms of atherosclerosis; unstable angina, myocardial infarction and stroke. Together, the family of MMPs can degrade all of the components of the blood vessel extracellular matrix and their activity therefore, is tightly regulated in normal blood vessels. The increased MMP activity during atherosclerotic plaque development and instability must therefore be caused by increased cytokine and growth factor-stimulated gene transcription, elevated zymogen activation and an imbalance in the MMP:TIMP ratio. It is therefore conceivable that inhibition of MMPs or re-establishing the MMP:TIMP balance may be useful in treating the symptoms of atherosclerosis. Recent studies using synthetic MMP inhibitors and gene therapy have highlighted the potential of such an approach. PMID- 11060723 TI - Therapeutic potential of ribozymes in haematological disorders. AB - Ribozymes are RNA molecules that possess the ability to cleave and thus destroy other RNA molecules. As a result of this ability, they are ideal specific agents to use against the messenger RNAs of important genes found to be linked with disease (of cellular and viral origin). This review will briefly describe the different types of ribozyme and the potential they have as therapeutic compounds against viruses, oncogenes and drug resistance in haematological settings. The latest news from the various Phase I and II ribozyme clinical trials is discussed, as is the potential for the ribozymes' future as therapeutic agents. PMID- 11060724 TI - Gene therapy and heart transplantation. AB - The application of gene transfer technologies to the field of solid organ transplantation is uniquely appealing due to open access to the donor organ at the time of removal and the need for a local biological effect limited to the allograft. The objectives of gene transfer technology in the field of experimental heart transplantation include: firstly, modification of allograft phenotype and secondly, modulation of the host alloimmune response. Both objectives can theoretically decrease or eliminate the need for lifelong immunosuppression with its attendant risks. This article will review the principles and current methodology of gene transfer technology, applications of gene transfer technology to allo- and xeno- transplantation and the current status of clinical trials on gene therapy. PMID- 11060725 TI - Gene therapy for kidney disease. AB - Gene therapy has distinct potential to treat disease at the most fundamental level. However, the ability to pursue gene therapy for renal disease has been limited by the availability of an adequate system for gene delivery to the kidney and for regulation of transgene expression. Presently, there are several limitations to overcome before clinical use of viral vector systems for targeting kidney can be considered. Non-viral vectors such as haemagglutinating virus of Japan (HVJ)-liposome mediated gene transfer and cationic liposome are promising but need to be improved. Given that the systemic delivery of the functional protein can serve as therapy for the renal diseases, skeletal muscle targeting gene therapy might be an alternative strategy for the treatment of renal disease. Gene therapy to the transplant kidney may potentially improve the graft outcome by reducing acute and chronic rejection. We review emerging strategies of gene transfer with reference to the kidney and discuss the potential application of gene therapy to renal diseases. PMID- 11060726 TI - The renoprotective potential of endothelin receptor antagonists. AB - The endothelin system has been identified as having a substantial role in renal failure, both acute and chronic. Beside its well characterised haemodynamic effects, its mitogenic and pro-fibrotic properties have gained increased interest in the pathophysiology of chronic renal failure. This review outlines the role of endothelin in the pathogenesis of various renal diseases with a special focus on the potential of blocking this system with endothelin receptor antagonists. So far, most data were derived from animal models, but they provide strong evidence that endothelin receptor antagonists may represent a powerful therapeutic strategy in ameliorating the course of acute and chronic renal failure. PMID- 11060727 TI - Cyclosporin treatment of glomerular diseases. AB - Cyclosporin is a potent immunosuppressive agent that has become the first line therapy in organ transplantation. Its efficacy has led to its use in a variety of immune-mediated glomerular diseases. A selection of controlled and uncontrolled trials has studied the effects of cyclosporin in patients with minimal change disease (MCD), focal segmental glomerulosclerosis (FSGS), membranous nephropathy (MN), IgA nephropathy, membranoproliferative glomerulonephritis (MPGN) and lupus nephritis. We review the recent literature and suggest recommendations for using cyclosporin in these diseases, based on this evidence and our experience. PMID- 11060728 TI - Necatoriasis: treatment and developmental therapeutics. AB - Two hookworm parasites, Necator americanus and Ancylostoma duodenale, infect approximately one billion people worldwide. These hookworms are one of the leading causes of iron-deficiency anaemia especially in children, resulting directly from intestinal capillary blood loss following the feeding activities of fourth-stage (L(4)) larva and adult worms. If ignored, human hookworm infections can retard growth and the intellectual development of children. Another clinical manifestation often associated with hookworm infections is cutaneous larva migrans (CLM). It is a well recognised, usually self-limiting condition caused by the infectious larvae of nematodes, especially Ancylostoma spp. CLM is characterised by skin eruption and represents a clinical description rather than a definitive diagnosis. Of the hookworm parasites, the dog and cat worm A. braziliense and A. caninum are the most common nematodes causing CLM, although many other species have also been implicated. The major subject of this review article will be discussion of the evolution of therapies and treatment of human necatoriasis and the development of experimental infections with N. americanus. Difference in the clinical efficacy of mebendazole and albendazole will be discussed along with drug resistance of N. americanus. PMID- 11060729 TI - Chemokine receptor antagonism as a new therapy for multiple sclerosis. AB - New information about the role of tissue inflammation in the pathogenesis of multiple sclerosis (MS) has driven a search for effective and specific therapeutics that address leukocyte trafficking. These developments in understanding MS are complemented by advances in clarifying the molecular mechanisms of leukocyte extravasation and providing the knowledge base needed to modulate tissue inflammation. Of particular interest are the chemokines and their receptors. Chemokines constitute a large family of chemoattractant peptides that regulate the vast spectrum of leukocyte migration events. This review discusses MS and proposes that identifying the chemokines and receptors involved in the inflammation associated with this disorder may lead to therapeutic intervention. PMID- 11060730 TI - Pharmacology and clinical assessment of cariporide for the treatment coronary artery diseases. AB - Myocardial protection through pharmacological approaches represents a large therapeutic challenge and is an important therapeutic strategy in patients with coronary artery disease, particularly after myocardial infarction. Extensive animal experiments have repeatedly demonstrated the efficacy of sodium-hydrogen exchange (NHE) inhibition as a potent cardioprotective approach. The heart possesses primarily the NHE1 isoform which has led to the development of NHE1 specific inhibitors for cardiovascular therapeutics. Cariporide (HOE 642) is the first of such agents to have been developed and subjected to clinical trial. Preclinical studies with cariporide revealed excellent protection against necrosis, apoptosis, arrhythmias and mechanical dysfunction in hearts subjected to ischaemia and reperfusion. Cariporide has recently been evaluated in a large dose-finding Phase II/Phase III clinical trial (GUARDIAN) to assess its efficacy in patients with acute coronary syndromes. Overall results failed to demonstrate protection but sub-group analysis revealed significant risk reductions with the highest cariporide dose (120 mg t.i.d.) especially in high risk patients undergoing coronary artery bypass surgery. This suggests that insufficient dosage may have accounted, at least in part, for the less than optimum results. Another NHE1 inhibitor, eniporide, is currently in Phase II clinical trial (ESCAMI) in patients with acute myocardial infarction (MI) who are given angioplasty or thrombolysis. Although the study has not been completed interim findings appear positive. Both drugs were well-tolerated and produced no excess side effects compared with placebo. Further studies are needed to confirm the efficacy of NHE1 inhibitors for the treatment of coronary heart disease, even so initial results are encouraging. PMID- 11060731 TI - Clinical overview of the novel inotropic agent toborinone. AB - Toborinone (OPC-18790, Otsuka Pharmaceutical Co. Ltd, 2(1H) -quinolone,6-[3-[ [3,4-dimethoxyphenyl)methyl] amino]-2-hydroxy prop oxyl]-,(.+-.)-) is a novel iv. inotropic agent. Positive inotropic effects are produced by PDE inhibition with the resulting increase in cAMP and intracellular calcium levels. Unlike other inotropic agents that increase cAMP, there is an absence of positive chronotropic effects, which are attributed to prolongation of the action potential due to blockade of delayed rectifier currents. There is also marked venous and arterial vasodilating properties. The absence of heart rate increases results in decreased myocardial oxygen consumption compared with conventional inotropes. Studies in human heart failure patients have been consistent with previous work in animal studies, confirming the effects of toborinone as being positive inotropy (relatively weak), marked arterial and venous vasodilatation and absence of increase in myocardial oxygen consumption. Data regarding safety in larger clinical trials, particularly regarding arrhythmias, is at present unavailable. This information will determine whether this agent becomes an accepted iv. therapeutic option for congestive heart failure. PMID- 11060732 TI - Bivalirudin: a new generation antithrombotic drug. AB - Bivalirudin (Angiomax, The Medicines Company) is a synthetic 20 amino acid peptide rationally designed on the basis of structural studies of hirudin, a naturally occurring anticoagulant. Bivalirudin represents a new class of anticoagulant drugs that directly inhibits thrombin, a key component in blood clot formation and extension. With its high binding affinity and specificity for thrombin, bivalirudin acts directly on thrombin, rather than via other clotting factors. The compound has a variety of potential uses as an alternative to heparin in the management of cardiovascular disease and related medical procedures i.e., unstable angina (UA), myocardial infarction (MI) and percutaneous transluminal coronary angioplasty (PTCA). PMID- 11060733 TI - Effects of melagatran, a novel direct thrombin inhibitor, during experimental septic shock. AB - Sepsis and endotoxaemia initiate the generation of thrombin, which is responsible for the conversion of fibrinogen to fibrin, platelet aggregation and acts as an inflammatory mediator affecting numerous types of cells, including myocardial, smooth muscle and endothelial cells. Human Gram-negative septic shock, frequently seen in intensive care units, is a condition with high mortality. This condition can be replicated in the endotoxaemic pig. As many of the toxic effects of sepsis are due to thrombin generation, it was of interest to study, using this porcine experimental septic shock model, whether inhibition of thrombin could alleviate the effects of endotoxaemia. For this purpose melagatran, a direct synthetic thrombin inhibitor with a molecular weight of 429 Da, was employed. Melagatran does not significantly interact with any other enzymes in the coagulation cascade or fibrinolytic enzymes aside from thrombin. Furthermore, melagatran does not require endogenous co-factors such as antithrombin or heparin co-Factor II for its antithrombin effect, which is important, as these inhibitors are often consumed in septic patients. We have shown that melagatran exerts a beneficial effect on renal function, as evaluated by plasma creatinine and urinary output, during experimental septic shock. These effects were most pronounced during the later phase of the experimental period, after the infusion of melagatran had been discontinued. Prevention of intrarenal coagulation may be attributable to this finding. In addition, melagatran had beneficial effects on systemic haemodynamics (left ventricular stroke work index, pulmonary capillary wedge pressure and systemic vascular resistance index) in endotoxaemic pigs. This result may be explained by the ability of melagatran to inhibit thrombin, thereby counteracting thrombin's cellular effects. Thus, it can be seen, using this experimental model of septic shock, that melagatran may help to alleviate some of the damaging effects of endotoxaemia, although more research is required to test this further. PMID- 11060734 TI - Evaluation of mitoxantrone for the treatment of multiple sclerosis. AB - Mitoxantrone (Novantrone((R))), an antineoplastic agent, has been approved for treating patients with secondary progressive multiple sclerosis (MS). Mitoxantrone, which is usually categorised as an immunosuppressant drug, is now also considered to be a specific immunomodulator. Autoimmune mechanism of pathogenesis of MS is the basis of immunosuppressive therapeutic approaches to MS whereas immunoregulatory abnormalities including defective IFN-alpha production provide the rationale for immunomodulating therapies. Clinical trials have shown that mitoxantrone had a statistically significant impact on reduction of relapse rate and delay in disability progression in these patients. Advantages of mitoxantrone as therapy for MS are: (1) considerable information is available about its pharmacokinetics, metabolism and toxicology from previous use in oncology; (2) it requires administration only once in three months which is not only convenient for the patient but also cost-effective; (3) mitoxantrone is one of the two drugs to be approved for secondary progressive MS (the other is IFN beta1) which offers an advantage over IFN-beta1a preparations and glatiramer acetate which are indicated only for relapsing remitting MS. However, the duration of therapy is usually limited to two to three years because the maximum cumulative dose recommended is 120 mg/m(2) due to concern for possible cardiotoxicity. Potential market value of the mitoxantrone, based on the cost of treatment per patient and the number of patients likely to be treated in the first year of introduction, is US$210 million. PMID- 11060735 TI - Novel non-operative treatment and treatment strategies in pancreatic cancer. AB - Patients with advanced pancreatic cancer have traditionally been treated with palliative care only. The last decade has seen significant improvements in the surgical treatment of this disease but until the late 1990s there was no effective non-surgical treatment for these tumours. The introduction of gemcitabine has given clinicians treating patients with pancreatic cancer a new option. The published randomised data of gemcitabine in patients with pancreatic cancer has shown both a small survival advantage and significant improvements in quality of life indicators in these patients. These data have stimulated a resurgence of interest in pancreatic tumours and several studies have been or are currently investigating novel treatments or treatment strategies. The explosion in the molecular knowledge of cancer has led to the development of several 'molecular designer drugs' that have been tested in pancreatic cancer. The furthest advanced of these is a matrix metalloproteinase (MMP) inhibitor called marimastat. The first randomised data using this new class of agents is increasing and suggests that marimastat may have a role in the future treatment of patients with pancreatic cancer. Other agents such as gastrimmune, are about to enter Phase III studies and several other molecular treatment strategies are progressing from the in vitro stage towards the clinical arena. Each of these treatments and treatment regimens are discussed along with their current progress. PMID- 11060736 TI - Targeted therapies for the myeloid leukaemias. AB - Although the standard approach to myeloid leukaemias remains chemotherapy, the agents currently available rarely result in cure. Recent advances in understanding the biology of these disorders have lead to the development of targeted treatment strategies. In acute promyelocytic leukaemia (APL), all-trans retinoic acid (ATRA), sodium phenylbutyrate and arsenic trioxide are agents which either induce differentiation or apoptosis and have been used to successfully achieve remission. The tyrosine kinase inhibitor, STI-571, antisense oligonucleotides, and bcr-abl vaccines are strategies which focus on the oncogenic events in chronic myelogenous leukaemia (CML). Two anti-CD33 monoclonal antibody conjugates, Y90-HuM195 and CMA-676, have been used in acute myelogenous leukaemia (AML) and have shown some efficacy. Although the preliminary results with these targeted therapies are promising, further studies are needed to establish them as effective, less toxic alternatives to the current standard of care. PMID- 11060737 TI - Molecular advances to treat cancer of the brain. AB - Malignant primary and metastatic brain tumours continue to be associated with poor prognosis. Nevertheless, recent advances in molecular medicine, specifically in the strategies of gene therapy, targeting tumour cells, anti-angiogenesis and immunotherapy, have created novel tools that may be of therapeutic value. To date, gene therapy trials have not yet demonstrated clinical efficacy because of inherent defects in vector design. Despite this, advances in adenoviral technology, namely the helper-dependent adenoviral constructs (gutless) and the uncovering of brain parenchymal cells as effective and necessary targets for antitumour benefits of adenoviral-mediated gene transfer, suggest that developments in vector design may be approaching the point of clinical utility. Targeting tumour cells refers to strategies that destroy malignant but spare normal cells. A new assortment of oncolytic viruses have emerged, capable of specific lysis of cancer tissue while sparing normal cells and propagating until they reach the tumour borders. Furthermore, peptides have been transformed into bullets that specifically seek and destroy cancer cells. The concept of tumour angiogenesis has been challenged by new but still very controversial findings that tumour cells themselves may form blood channels. These results may lead to the redirecting of the molecular targets toward anti-angiogenesis in some tumours including glioblastoma multiform. Unfortunately, our knowledge regarding the immunological ignorance of the tumour is still limited. Even so, newly discovered molecules have shed light on novel pathways leading to the escape of the tumour from the immune system. Finally, significant limitations in our current experimental tumour models may soon be overcome by firstly, the development of models of reproducible organ-specific tumours in non-inbred animals and secondly applying genomics to individualize therapy for a particular tumour in a specific patient. PMID- 11060738 TI - Prostate cancer immunotherapy at the dawn of the new millennium. AB - Standard treatments for adenocarcinoma of the prostate, such as surgery, hormones, radiation and chemotherapy, often achieve a clinical response, but this is usually short-lived. Prostate cancer frequently recurs and second-line therapies have a poor response rate. Many clinicians seem comfortable in limiting their philosophy of treating advanced recurrent disease merely to new regimens of failed therapies, such as combination chemotherapy. However, other medical researchers have chosen to pursue novel approaches, including immunotherapy, several of which are summarised in this review. Although ranging widely in antigen specificity, all attempt to exploit the body's natural antitumour immunity. Furthermore, all aim to stimulate immunity above a threshold level necessary for tumour regression or to induce stability in the face of progression. The goal of in vivo or ex vivo gene therapy is the modification of gene expression within an antigen-presented cell by the introduction of a vector, DNA, or RNA. Within that field, much progress has been made and is ongoing currently concerning gene delivery systems, target identification and characterisation. Comparatively, monoclonal antibodies are an established type of cancer immunotherapy. However, the more recent development of humanized or fully human antibodies, as well as novel moieties they can be coupled to, renews their prospects for clinical impact. Lastly, various cell-based therapies are the focus of several recent clinical studies demonstrating tumour regression or stabilisation. Immune cells, for example, T-lymphocytes and dendritic cells, have already demonstrated treatment benefit, as well as the ability to maintain an excellent quality of life for participants. Overall, there is a multitude of approaches being considered for the treatment of prostate cancer. The following review concentrates on those approaches that are currently in human or animal studies and have a specific emphasis on prostate cancer. PMID- 11060739 TI - Recombinant antibodies: a novel approach to cancer diagnosis and therapy. AB - Recombinant antibodies and their fragments currently represent over 30% of all biological proteins undergoing clinical trials for diagnosis and therapy. These reagents dominate the cancer-targeting field, as highlighted by the recent approval of the first engineered therapeutic antibodies by the Food and Drugs Administration (FDA). Last year, important advances have been made in the design, selection and production of recombinant antibodies. The natural immune repertoire and somatic cell affinity maturation has been superseded by large antibody display libraries and rapid molecular evolution strategies. These novel libraries and selection methods have enabled the rapid isolation of high-affinity cancer targeting and antiviral antibodies, the latter capable of redirecting viruses for gene therapy applications. In alternative strategies for cancer diagnosis and therapy, recombinant antibody fragments have been fused to radioisotopes, drugs, toxins, enzymes and biosensor surfaces. Antibody-directed cancer pre-targeting followed by prodrug activation (ADEPT) has proved a most promising therapeutic strategy. Multi-specific antibodies have been effective for cytotoxic T-cell recruitment and antibody-fusion proteins have delivered enhanced immunotherapeutic and vaccination strategies. The new millennium is indeed an exciting time for the design, selection and formulation of a range of new antibody-based products for cancer diagnosis and therapy. PMID- 11060740 TI - Microbial pathogens and apoptotic anticancer therapy. AB - In spite of tremendous efforts to control cancer, the mortality associated with this disease has been increasing in developed countries in the recent decades. Inadequate efficiency of existing therapeutic regimens and the rise of multi-drug resistant cancer cells are the main factors which require a broadening of investigations into novel anticancer strategies. Enhancement of apoptosis in tumours has been suggested recently as a new anticancer strategy. It targets the disruption of equilibrium between cell proliferation and cell death in tumours and suggests to restore it through the use of pharmacological agents or genetic approaches. Apoptotic therapy has attracted many groups of investigators and several companies have entered the race to develop the first generation of apoptotic anticancer agents. The review discusses the role that pathogenic microorganisms may have as the source of agents for apoptotic therapy. PMID- 11060741 TI - Complexes of gallium(III) and other metal ions and their potential in the treatment of neoplasia. AB - The metal complexes of a variety of ligands show diverse pharmacological properties. The potential of these compounds as antineoplastic agents is underlined by the success of the clinically used platinum complex cisplatin (cis [(NH(3))(2)PtCl(2)]). In the current review, specific examples of gallium, copper, ruthenium and titanium complexes are discussed with special relevance to their use in the treatment of cancer. Some of these complexes have demonstrated marked activity in a number of animal models and for some compounds, clinical trials are anticipated or have already begun. Collectively, the results in the literature indicate that the study of metal complexes as antineoplastic agents deserves continued intensive investigation. PMID- 11060742 TI - Small molecule alpha(v) integrin antagonists: novel anticancer agents. AB - The members of the integrin family are targets that potentially provide both therapeutic and diagnostic opportunities. Advances in the understanding of the signalling pathways, transcriptional regulation and the structure/function relationships of the adhesion molecules to extracellular matrix proteins have all contributed to these opportunities. The role of the integrins in pathological processes in both acute and chronic diseases, include ocular, cancer (solid tumours and metastasis), cardiovascular (stroke and heart failure) and inflammatory (rheumatoid arthritis) conditions. Various therapeutic candidates, including antibodies, cyclic peptides and peptidomimetics, have been identified. This review will focus on the key role of the alpha(v) integrin (alpha(v)beta(3) and alpha(v)beta(5)) in angiogenic processes in tumours, including its potential use in cancer diagnostics and therapy. PMID- 11060743 TI - alpha(v)beta(3) Integrin antagonists as inhibitors of bone resorption. AB - The alpha(v)beta(3) integrin is a non-covalent, heterodimeric, cell-surface protein that is expressed with varying density on numerous cell types, including osteoclasts, vascular smooth muscle cells, endothelial cells and a variety of tumour cells. Functionally, alpha(v)beta(3) mediates a diverse range of biological events including the adhesion of osteoclasts to bone matrix, smooth muscle cell migration and angiogenesis. Specifically, there has been significant attention focused on the preparation of inhibitors of alpha(v)beta(3) for use as inhibitors of bone resorption, in recognition of the medical need for improved prevention and treatment of osteoporosis. Herein, we summarise the pertinent chemistry and biological advances in the medicinal design and biological evaluation of peptide and small molecule alpha(v)beta(3) antagonists as inhibitors of bone resorption. PMID- 11060744 TI - The parathyroid hormone, its fragments and analogues--potent bone-builders for treating osteoporosis. AB - As populations age a rising number of men and women, but especially women during the first decade after menopause, become victims of a severe, accelerated loss of bone with crippling fractures known as osteoporosis. This often results in costly, prolonged hospitalisation and perhaps indirectly, death. Osteoporosis in women is caused by the menopausal oestrogen decline, which removes several key restraints on the generation, longevity and activity of bone-resorbing osteoclasts. Although there are many antiresorptive drugs on or coming onto the market (calcitonin, bisphosphonates, oestrogen and SERMS) that can slow or stop further bone loss, there are none that can restore lost bone mechanical strength by directly stimulating osteoblast activity and bone growth. However, there is a family of potent bone-building peptides, namely the 84 amino acid parathyroid hormone (PTH). Its 31 to 38 amino acid N-terminal fragments are currently in or about to enter clinical trials. We can predict that these peptides will be effective therapeutics for osteoporosis especially when supplemented with bisphosphonates or SERMs to protect the new bone from osteoclasts. These peptides should also accelerate the healing of fractures in persons of all ages and restore lost bone mass and mechanical strength to astronauts following their return to earth after long voyages in space. PMID- 11060745 TI - Novel anti-obesity drugs. AB - There is increasing evidence that body weight is homeostatically regulated and that in obesity this regulation maintains weight at a high level. Weight loss activates mechanisms that are designed to return individuals to their pre existing weight. This explains the universally poor results of current strategies to maintain weight loss. On this basis, life-long drug therapy may be justified for those with significant obesity. Currently available drugs include selective serotonin re-uptake inhibitors (e.g., fluoxetine), noradrenergic re-uptake inhibitors (e.g., phentermine), a serotonin and noradrenergic re-uptake inhibitor (sibutramine) and an intestinal lipase inhibitor (orlistat). An active research program is underway to develop new agents based on the rapidly expanding knowledge of the complex mechanisms regulating body weight. Leptin, a hormone produced by adipocytes that inhibits food intake, has undergone clinical trials and analogues are currently being developed. Other agents include amylin, melanocortin-4 receptor agonists, neuropeptide Y antagonists, beta(3) adrenergic agonists and glucagon-like peptide-1 agonists. As some redundancy exists in the central regulatory system controlling body weight, some agents might need to be used in combination to be effective. PMID- 11060746 TI - The role of NPY in metabolic homeostasis: implications for obesity therapy. AB - Neuropeptide Y (NPY) is a 36 amino acid amidated peptide which has now emerged as an important regulator of feeding behaviour. Upon intracerebroventricular (icv.) administration, NPY produces a pronounced feeding response in a variety of species. The actions of NPY are believed to be mediated by a family of receptor subtypes named Y1 - y6. Recent studies suggest that the Y1 and Y5 receptor subtypes are intimately involved in NPY induced feeding. This review presents preclinical data obtained with receptor subtype selective agonists and antagonists as well as findings from knockout mice. These new data suggest that NPY receptor antagonists may become an additional option for treating human obesity. PMID- 11060747 TI - Insulin sensitiser drugs. AB - Insulin resistance is the predominant early pathological defect in Type 2 diabetes. As well as being a risk factor for the development of Type 2 diabetes, insulin resistance is also associated with increased cardiovascular risk and other metabolic disturbances including visceral adiposity, hyperinsulinaemia, impaired glucose tolerance, hypertension and dyslipidaemia [1-4]. The newest approach to oral antidiabetic therapy is to target improvements in insulin sensitivity at muscle, adipose tissue and hepatic level. This results in improvements in glycaemic control and other features of the insulin resistance syndrome, with potential long-term benefits in preventing/delaying the onset of diabetic complications and macrovascular disease. PMID- 11060748 TI - Novel agents in the therapy of endotoxic shock. AB - Endotoxic shock, or Gram-negative septic shock, can occur as a component of Gram negative sepsis and is characterised by hypotension, poor tissue perfusion and multi-organ dysfunction. Despite antibiotic therapy and intensive care management, the morbidity and mortality rates of Gram-negative septic shock remain high. Endotoxin mediates its effects through interaction with receptors on the surface of a variety of host cells. These interactions result in the production and release of numerous biochemical mediators including nitric oxide, cytokines, prostaglandins and leukotrienes and toxic oxygen radicals. It is these biochemical mediators that exert toxic effects during endotoxic shock and which are often the target of novel treatment strategies. Several of these pharmacological agents are currently being investigated for use in Gram-negative septic shock and include inhibitors of the enzyme responsible for nitric oxide production, scavengers of the nitric oxide molecule and cytokine modulators. Although many agents have been studied for potential use as modulators of cytokine levels, this study will focus on pentoxifylline and the 21 aminosteroids, or lazaroids. Examination of the literature regarding pharmacological agents used to treat endotoxic shock often yields confusing and contradictory results. The reasons for these mixed results include differences in models, drug dosages, dosing methods and intervals and timing of administration relative to disease duration and severity. However, despite mixed results, several of the drugs discussed in this paper offer promise in the therapy of an often frustrating and lethal condition. PMID- 11060749 TI - An update on satraplatin: the first orally available platinum anticancer drug. AB - This update focuses on the clinical development of the first orally available platinum-containing anticancer drug, satraplatin (JM216, BMS 182751, BMY 45594). Satraplatin was selected for clinical study on the basis of possessing several promising preclinical features the first of which is it's potent in vitro growth inhibitory properties against several tumour types (mean IC(50) approximately 1 microM). Secondly, it possesses in vivo oral antitumour activity against a variety of murine and human sc. tumour models, broadly comparable to the level of activity obtainable with parenterally administered cisplatin or carboplatin. Lastly, it has a relatively mild toxicity profile with myelosuppression being dose-limiting. Satraplatin entered clinical trials in 1992 and is now undergoing Phase III evaluation. Non-linear pharmacokinetics, probably due to saturable absorption, was observed when the drug was administered as a bolus every 3 - 4 weeks. Subsequent Phase II trials have used a daily schedule for five consecutive days, at doses of around 120 mg/m(2)/day. The drug produced relatively mild side effects with controllable nausea and vomiting and, as predicted from the mouse studies, myelosuppression as the dose-limiting effect (neutropoenia and thrombocytopoenia). Combination trials are also ongoing with paclitaxel or radiation. The metabolism of satraplatin is complex, with at least six biotransformation products observed in the plasma of patients. The platinum(II) complex JM118 is the main metabolite, three other minor metabolites have been identified, there is no detectable parent drug. Tumour responses have been recorded, particularly in patients with small cell lung cancer and hormone refractory prostate cancer. These clinical studies with satraplatin indicate that oral platinum-based chemotherapy is feasible. PMID- 11060750 TI - TNP-470: an angiogenesis inhibitor in clinical development for cancer. AB - TNP-470, an analogue of fumagillin, has been shown to inhibit angiogenesis in vitro and in vivo. In 1992, TNP-470 entered clinical development for cancer as an anti-angiogenic agent. It is currently in Phase I/II trials in Kaposi's sarcoma, renal cell carcinoma, brain cancer, breast cancer, cervical cancer and prostate cancer. In early clinical reports, TNP-470 is tolerated up to 177 mg/m(2) with neurotoxic effects (fatigue, vertigo, ataxia, and loss of concentration) being the principal dose limiting toxicity (DLT). Terminal half-life values are short and have shown intermittent and intrapatient variation (range: 0.05 - 1.07 h). Recently, mechanistic studies have identified cell cycle mediators and the protein methionine aminopeptidase-2 (MetAP-2) as molecular targets of TNP-470 and fumagillin. Animal studies confirm some toxic effects on normal angiogenic processes such as the female reproductive system and wound healing, which will require caution and close monitoring in the clinic. TNP-470 is one of the first anti-angiogenic compounds to enter clinical trials, making it a valuable prototype for future trials of angiogenesis inhibitors in oncology. PMID- 11060751 TI - Evaluation of memantine for neuroprotection in dementia. AB - Memantine, a non-competitive NMDA antagonist, has been approved for use in the treatment of dementia in Germany for over ten years. The rationale for use is excitotoxicity as a pathomechanism of neurodegenerative disorders. Memantine acts as a neuroprotective agent against this pathomechanism, which is also implicated in vascular dementia. HIV-1 proteins Tat and gp120 have been implicated in the pathogenesis of dementia associated with HIV infection and the neurotoxicity caused by HIV-1 proteins can be blocked completely by memantine. Memantine has been investigated extensively in animal studies and following this, its efficacy and safety has been established and confirmed by clinical experience in humans. It exhibits none of the undesirable effects associated with competitive NMDA antagonists such as dizocilpine. The efficacy of memantine in a variety of dementias has been shown in clinical trials. Memantine is considered to be a promising neuroprotective drug for the treatment of dementias, particularly Alzheimer's disease for which there is no neuroprotective therapy available currently. It can be combined with acetylcholinesterase inhibitors which are the mainstay of current symptomatic treatment of Alzheimer's disease. Memantine has a therapeutic potential in numerous CNS disorders besides dementias which include stroke, CNS trauma, Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), epilepsy, drug dependence and chronic pain. If memantine is approved by the FDA for some of these indications by the year 2005, it can become a blockbuster drug by crossing the US$1 billion mark in annual sales. PMID- 11060752 TI - Metalloproteinase inhibitors: new opportunities for the treatment of rheumatoid arthritis and osteoarthritis. AB - The clinical efficacy of anti-TNF-alpha therapies have highlighted the apparently central role that TNF-alpha plays in the pathology of rheumatoid arthritis, particularly the inflammatory component. Recent identification of a metalloproteinase from the metzincin superfamily responsible for the production of the soluble form of this cytokine, has generated a large amount of pharmaceutical interest and presents the prospect of a metalloproteinase inhibitor as an anti-inflammatory drug. However, the traditional focus of metzincin inhibitor research has been the identification of inhibitors of matrix metalloproteinases; enzymes associated with matrix destruction, a feature common to both rheumatoid arthritis and osteoarthritis. Inhibitors of this class of metalloproteinase are now in clinical evaluation in patients. This review summarises the current development status of metalloproteinase inhibitors in arthritic diseases and discusses some of the issues that have arisen during their progress to become clinical treatments for these diseases. PMID- 11060753 TI - Biological agents: a novel approach to the therapy of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is the most common chronic autoimmunopathy, clinically leading to joint destruction as a consequence of the chronic inflammatory processes. The pathogenesis of this disabling disease is not well understood, but molecular events leading to tissue inflammation with cartilage and bone destruction are now defined in more detail. Established therapy, slow acting disease-modifying antirheumatic drugs (DMARDs) as with low-dose methotrexate (MTX) are the accepted 'golden standard' therapies and both lead to a significant improvement of disease symptoms, however are unable to stop joint destruction. Due to these disappointing treatment options and the identification of some inflammatory mediators as therapeutic targets, novel therapeutic agents such as monoclonal antibodies (mAbs), cytokine receptor-human immunoglobulin constructs or recombinant human proteins have been tested in RA with some success. In particular, clinical trials testing anti-TNF-alpha agents either alone or in combination with MTX have convincingly demonstrated the feasibility and efficacy of these novel approaches to the therapy of RA. Importantly, a clinical trial testing combination therapy with chimeric (mouse-human) anti-TNF-alpha mAb cA2 (Remicadetrade mark) and MTX could, for the first time in any RA trial, show that average radiological progression in the cA2/MTX groups could be completely prevented over a 12 month observation period. Similar encouraging results might evoke from trials employing other TNF-alpha-directed agents like the fully human mAb D(2)E7 or the p75 TNF-alpha-receptor-Ig construct, etanercept. PMID- 11060754 TI - Tetracyclines for the treatment of rheumatoid arthritis. AB - In this review the rationale for the possible beneficial effect of tetracycline derivatives for the treatment of rheumatoid arthritis is discussed. Early studies (Sanchez, Skinner et al. and Brown et al. ) and the two open trials of the 1980s are briefly discussed. The three double-blind studies conducted in the 1990s (Kloppenburg et al. , The Netherlands; the MIRA trial, USA and the O'Dell et al., USA) are described in detail. The baseline clinical and demographic data for these patients, as well as the efficacy and toxicity data are described in the text and summarised in tables. The long-term data of the O'Dell et al.'s trial is described. Finally, side effects not observed during the conduct of these trials, but reported to occur in other patients, for example those receiving minocycline for the treatment of acne, are also listed. PMID- 11060755 TI - Novel strategies for the treatment of osteoarthritis. AB - Osteoarthritis is a worldwide heterogeneous group of conditions that leads to joint symptoms, which are associated with defective integrity of articular cartilage, in addition to related changes in the underlying bone at the joint margins. The prevalence of the disease after the age of 65 years, is about 60% in men and 70% in women. The aetiology of osteoarthritis is multifactorial, with the end result being mechanical joint failure and varying degrees of loss of joint function. The pathophysiological events associated with osteoarthritis are beginning to be understood. Essential inflammatory cytokines, such as IL-1beta and TNF-alpha, are involved initiating a vicious cycle of catabolic and degradative events in cartilage, mediated by metalloproteinases, which degrade cartilage extracellular matrix. The role of inflammation in the pathophysiology and progression of early osteoarthritis is supported further by the observation that C-reactive protein levels are raised in women with early knee osteoarthritis and higher levels predict those whose disease will progress. The synovium from osteoarthritis joints stains for IL-1beta and TNF-alpha. Nitric oxide, which exerts pro-inflammatory effects, is released during inflammation. Cartilage from patients with rheumatoid arthritis and osteoarthritis spontaneously produces nitric oxide in vitro. In experimental osteoarthritis, nitric oxide induces chondrocyte apoptosis, thus contributing to cartilage degradation. Hence unregulated nitric oxide production in humans plays a part in the pathophysiology of the disease. These recent observations suggest that therapy can now be targeted at specific sites of pathophysiological pathways involved in the pathogenesis of osteoarthritis. The novel strategies under consideration for the treatment of osteoarthritis can be divided into five main areas. These are COX-2 inhibitors, nitric oxide synthesis inhibitors and anti-oxidants, chondrocyte and bone growth promoters, metalloproteinase and cytokine inhibitors and gene therapy. PMID- 11060756 TI - Psoriatic arthritis. AB - Psoriatic arthritis occurs in 5 - 42% of patients with psoriasis. It is an inflammatory arthritis distinct from rheumatoid, being usually sero-negative, asymmetrical and often affecting the spine, sacro-iliac and distal interphalangeal joints. It runs a very variable course, from a mild non destructive disease to a severe rapidly progressive erosive arthropathy, producing an 'arthritis mutilans' with a combination of bone lysis and joint ankylosis. Its pathogenesis is not as well understood as rheumatoid arthritis, but is thought to be similarly immune driven, with a qualitatively similar immunomodulatory cascade and cytokine profile. Quantitatively, however, there are distinct differences in cell ratios and cytokine levels that may well impact on therapeutic strategies. Current therapies, such as methotrexate and sulphasalazine, have yet to be shown to be significantly more effective than placebo in delaying damage and produce only marginal improvements in symptoms. The newer specific biological agents, such as the anticytokine antibodies, interleukins and more specific anti-T-cell therapies, are starting to be studied in psoriatic arthritis. The rationale for their use comes mostly from extrapolation of their efficacy in rheumatoid arthritis. It has yet to be seen whether they will be efficacious in treating the osteolysis, fibrosis and new bone formation particular to psoriatic arthritis. Any treatment for the arthritis must also help the skin. Greater understanding of psoriatic arthritis, its pathogenesis and natural history is required if we are to target these exciting but expensive therapies effectively. PMID- 11060757 TI - Gene therapy for cystic fibrosis. AB - The gene for cystic fibrosis was identified in 1989 and this together with the emerging technology of gene therapy heralded a new dawn for the treatment of genetic disease. The initial optimism however gave way to the realisation that gene therapy for cystic fibrosis was unlikely to be straightforward. The lung was considered an ideal organ to target due to ease of access, but subsequent research has shown that the airway surface provides an efficient barrier to topically applied gene transfer agents. A number of Phase I clinical safety trials were carried out through the 1990s and provided proof of concept evidence that delivery of DNA by either viral or non-viral means was safe though not clinically efficacious. Current research is now focusing more on the barriers faced by delivery agents, with the aim that more efficient gene delivery will lead to a gene therapeutic for cystic fibrosis. PMID- 11060758 TI - Novel therapeutic approaches to gastric and duodenal ulcers: an update. AB - Over the last 25 years, a remarkable revolution in the pathophysiology and treatment of gastric and duodenal ulcers has occurred. Effective therapies were developed not only to heal ulcers, but also to cure most patients. The two principal causes for gastric and duodenal ulcers are either infection with Helicobacter pylori or the use of non-steroidal anti-inflammatory drugs (NSAIDs). With H. pylori eradication, gastric and duodenal ulcers are rapidly becoming historical diseases. This communication reviews the salient pharmacology of the novel anti-ulcer drugs currently in development, with particular emphasis on the treatment of gastric and duodenal ulcers. Intense research is currently focused on the development of proton pump inhibitors primarily for the treatment and prevention of gastroesophageal reflux disease. The older proton pump inhibitors, omeprazole and lansoprazole, are effective in healing gastric and duodenal ulcers. Furthermore, both drugs are effective in eradicating H. pylori when given with various antibiotics. Pantoprazole, rabeprazole and esomeprazole are new proton pump inhibitors, which appear to have comparable therapeutic profiles with omeprazole and lansoprazole. Rebamipide is a new mucosal protective drug, which is effective in healing gastric ulcers. Polaprezinc and nocloprost are also mucosal protective drugs, which are in clinical development. However, none of these three cytoprotective drugs have been evaluated for their efficacy in eradicating H. pylori when given in combination with antibiotics. Likewise, no published literature exists on the use of these drugs for preventing NSAID induced ulcers. With the rapid eradication of H. pylori currently happening in the developed world, the therapeutic challenge is now directed toward preventing NSAID-associated ulcer. Significant reduction of NSAID-induced ulcers is achieved by using continuous prophylactic anti-ulcer therapy (misoprostol or omeprazole) or by using NSAIDs possessing selective COX-2 inhibitory activity. However, outcome clinical studies are needed to compare the adjuvant anti-ulcer therapies given with COX-1 inhibitors versus the selective COX-2 inhibitors given alone. PMID- 11060759 TI - New agents in the treatment of soft-tissue sarcomas. AB - The list of agents with significant activity in soft-tissue sarcomas is very short and arguably, only includes doxorubicin and ifosfamide. Several other agents such as dacarbazine, cisplatin and etoposide, to name but a few, have marginal activity and are sometimes used in combination regimes. The need for the identification of new agents with activity against this disease is therefore paramount. Soft-tissue sarcomas are very rare and diverse and as a result, new drug trials are few and far fetched. The conventionally conducted Phase II trials, which include all histologies of soft-tissue sarcomas, run the risk of an inadequate assessment of the new drug in individual subsets, which may have variable biological behaviours. On the other hand, histology-specific Phase II trials are fraught with the problems of length due to the infrequent nature of the disease and the considerable running costs. The end result is that in this era of cost-containment, soft-tissue sarcomas are not high on the priority list of many funding agencies, thus explaining the lack of any significant progress over the past two decades. Efforts at optimising the dose intensity and schedule of administration of doxorubicin and ifosfamide, have resulted in superior activity when combined and improvement in the survival of patients with high-risk localised disease. However, efforts at identifying new agents with adequate activity have not had much success. Patients, physicians and pharmaceutical industries must be encouraged to participate in multidisciplinary clinical trials in order to improve cure rates for patients with advanced disease. PMID- 11060760 TI - Cannabinoid receptor ligands: clinical and neuropharmacological considerations, relevant to future drug discovery and development. AB - This review highlights some important advances that have taken place in cannabinoid research over the last four years. It focuses on novel ligands that are of interest either as experimental tools or as lead compounds for therapeutic agents and possible clinical applications for some of these ligands. The molecular targets for these compounds are various components of the system of endogenous cannabinoids (endocannabinoids) and receptors that together constitute the 'endocannabinoid system'. These are CB(1) cannabinoid receptors that are present mainly on central and peripheral neurones, CB(2) cannabinoid receptors that are expressed predominantly by immune cells, the biochemical mechanisms responsible for the tissue uptake or metabolism of endocannabinoids and vanilloid receptors. Other cannabinoid receptor types may also exist. Recently developed ligands include potent and selective agonists for CB(1) and CB(2) receptors, a potent CB(2)-selective antagonist/inverse agonist and inhibitors of endocannabinoid uptake or metabolism. Future research should be directed at characterising the endocannabinoid system more completely and at obtaining more conclusive clinical data about the possible beneficial effects of cannabinoids as well as their adverse effects. There is also a need for improved cannabinoid formulations/modes of administration in the clinic and advances in this area should be facilitated by the recent development of a potent water-soluble CB(1)/CB(2) receptor agonist. A growing number of strategies for separating sought-after therapeutic effects of cannabinoid receptor agonists from the unwanted consequences of CB(1) receptor activation are now emerging and these are discussed at the end of this review. PMID- 11060761 TI - Novel approaches to fracture healing. AB - The fractures that occur as a result of trauma frequently require multiple stage surgical procedures to achieve adequate union. Bone grafting with autogenous cancellous or cortico cancellous bone grafts is the traditional method used to repair bone defects. Most fractures will heal using this traditional procedure, however a number of fractures, up to 10% of the cases in United States alone, will result in delayed or impaired healing. Novel approaches are currently being investigated for the augmentation and acceleration of fracture healing. Some of these approaches include the use of biodegradable matrices; cell based approaches supplemented with osteogenic factors and genetic therapy. Cell based approaches for fracture healing have roused intense interest because of the great advance in the isolation and expansion of cells from the marrow that have the ability to differentiate into various types of cells including osteoblasts. In addition, the discovery and cloning of several proteins (bone morphogenetic proteins) that have the ability to induce bone formation, have contributed to the investigation of novel approaches to augment fracture healing. Use of genetic therapy for the augmentation of fracture healing has also recently gained strong interest. The attractive feature of gene therapy is that therapeutic proteins can be delivered locally to the fracture site in relatively high concentrations and in a sustained fashion. This review discusses these novel approaches and presents an assessment of their future clinical applicability. PMID- 11060762 TI - Therapy of systemic lupus erythematosus: new agents and new evidence. AB - Systemic lupus erythematosus (SLE) is a disease of relatively low prevalence with a wide range of clinical manifestations. Due in part to these two facts, there is little new evidence on the treatment of lupus. In fact, randomised controlled studies and prospective series are few and usually involve a small number of patients. Despite this, some therapies have shown to be beneficial within the last five years, while others emerge as possibilities in the near future. Among the former, antimalarials appear to be the treatment of choice for maintaining mild to moderate disease in remission. Methotrexate may be an alternative to other corticosteroid-sparing drugs, especially in patients with active arthritis and skin disease. Cyclosporin can be of use in proteinuric nephritis, although the incidence of hypertension with this drug is high. Thalidomide is useful for refractory skin lesions, but the efficacy of lower, less toxic doses is still to be studied. Immunoglobulins should probably be limited to selected patients with manifestations such as thrombocytopoenia. Experience is more limited with cladribine, fludarabine, tacrolimus, danazol and pentoxifylline. New therapies for severe SLE include mycophenolate mofetil, a potent immunosuppressive drug with a reasonable safety profile and immunoablative therapy with or without stem cell transplantation, in highly resistant cases or those with a poor prognosis. Other recently developed molecules, including anti-CD40L monoclonal antibodies (mAbs), are still under investigation. PMID- 11060763 TI - Recent advances in clinical trials of the direct and indirect selective Factor Xa inhibitors. AB - Over the years, pharmacological intervention to prevent undesired intravascular coagulation and the associated detrimental effects has been a clinical challenge. The first generation of anticoagulant agents, warfarin and unfractionated heparin (UFH), involve indirect mechanisms of inhibiting the coagulation cascade. Fractionated, or low-molecular weight, heparins (LMWHs) are more selective for coagulation Factor Xa (FXa) over thrombin (FIIa). LMWHs also utilise an indirect mechanism of inhibition and have improved pharmacokinetic, pharmacodynamic and therapeutic profiles over UFH. The success of LMWHs, along with the pivotal location of FXa in the coagulation cascade, has prompted interest in the discovery and development of selective FXa inhibitors. There are two general classes of FXa inhibitors in development, of which SR90107A/ORG31540, an antithrombin-III-dependent pentasaccharide and DX-9065a, a small molecule direct FXa inhibitor, have published clinical data. SR90107A/ORG31540 and DX-9065a offer safe, predictable pharmacokinetic and pharmacodynamic profiles when administered subcutaneously and intravenously, respectively, to healthy volunteers and appear to be progressing through clinical development. The purpose of this review is to compile and summarise the published Phase I and II clinical data for SR90107A/ORG31540 and DX-9065a. PMID- 11060764 TI - Inhaled nitric oxide in the neonatal period. AB - Pulmonary hypertension of the newborn is an important cause of hypoxaemia, particularly in the at term or near term neonate. It can occur as a primary condition or secondary to a variety of other diseases. Endogenous nitric oxide is an important modulator of vascular tone in pulmonary circulation. Initial uncontrolled studies indicated that inhalation of nitric oxide resulted in a reduction in pulmonary hypertension, with improvement in oxygenation, but no change in the systemic vascular resistance. There have now been a number of randomised trials performed exploring the efficacy of inhaled nitric oxide. These trials have demonstrated that in at term or near term infants, inhaled nitric oxide reduces the combined end point of death or the need for extracorporeal membrane oxygenation. The significant effect seems due to the reduced extracorporeal membrane oxygenation requirement. No such beneficial effect has been consistently reported in infants with congenital diaphragmatic hernia. Randomised trials have failed to highlight long-term positive results in preterm infants. Inhaled nitric oxide has side effects, although those due to nitrogen dioxide and methaemoglobin formation can be minimised by appropriate nitric oxide delivery. It is important to use the smallest effective nitric oxide dose, continuous nitric oxide and nitrogen dioxide monitoring and frequent methaemoglobin analyses. Careful patient selection should be undertaken, avoiding those at high risk of haemorrhagic complications. Longer term follow-up studies are required to determine the real risk:benefit ratio of inhaled nitric oxide treatment. PMID- 11060765 TI - An assessment of levetiracetam as an anti-epileptic drug. AB - A brief review of epilepsy as a disease, anti-epileptic drugs (AEDs) and methods of evaluation of AEDs are presented as a background for the assessment of levetiracetam which has been approved by the FDA as add-on therapy for the treatment of partial seizures with or without secondary generalisation in adults. The exact mechanism of action of levetiracetam is not known but its action differs from that of other anti-epileptic drugs. A specific binding site for levetiracetam has been identified and is possibly related to anticonvulsant activity. Levetiracetam offers an effective and broad spectrum treatment of epileptic seizures, partial as well as generalised epilepsy. Levetiracetam has been shown to be effective in genetic and kindled animal models of epilepsy and against chemoconvulsant-induced partial epileptic seizures. Levetiracetam has a near perfect pharmacokinetic profile, with rapid absorption following oral administration, excellent bioavailability, quick attainment of steady-state concentrations, linear kinetics and minimal plasma protein binding. Levetiracetam does not interact with commonly used drugs and other AEDs. In recent Phase III clinical trials, the responder rate was 39.4 - 42.1% on 3000 mg dose, compared with placebo rates of 10.9 - 16.7%. Levetiracetam has a favourable safety profile and the most frequently reported adverse events were somnolence, asthenia and dizziness. Overall, levetiracetam is considered to have several advantages over current AEDs. PMID- 11060766 TI - D-Sotalol: death by the SWORD or deserving of further consideration for clinical use? AB - D-Sotalol is the dextro-rotatory isomer of sotalol and a class III anti arrhythmic. D-Sotalol prolongs cardiac repolarisation by inhibiting the fast component of the delayed outward rectifying potassium channel. In animal studies, D-sotalol has been shown to be more effective in prolonging atrial, rather than ventricular, action potentials, suggesting that D-sotalol may be more effective against supra-ventricular than ventricular arrhythmias. Furthermore, in animal studies, D-sotalol induces after-depolarisations, which are predictors of pro arrhythmic activity. D-Sotalol shows little or no reverse use dependence in animal and humans and has slow offset kinetics. This suggests that, in addition to being a preventative treatment for arrhythmias, D-sotalol may be effective at the start or during arrhythmia. As D-sotalol does not block the slow component of the delayed outward rectifying potassium channel, which is activated by the sympathetic nervous system, D-sotalol will not protect against sympathetic hyperactivity. D-Sotalol also has no effect on the K(ATP) channel, which is activated in ischaemia to shorten the action potential. Thus D-sotalol is less effective in ischaemia. Anti-arrhythmic activity with D-sotalol has been demonstrated in dog models of ventricular tachycardia and sudden death. Arrhythmias with D-sotalol have been demonstrated in an ischaemic guinea-pig ventricle model in the absence of action potentials. D-Sotalol is a weak beta adrenoceptor antagonist and may also be a positive inotrope. In humans, D-sotalol has 100% systemic oral bioavailability, a terminal half-life of 7.2 h and is mainly excreted unchanged in the urine. Preliminary, mainly hospital-based, clinical trials showed that D-sotalol was effective in a variety of supraventricular and ventricular arrhythmias. However, a large clinical trial of D-sotalol as a preventative treatment for arrhythmias and sudden death after myocardial infarction, the SWORD trial, was terminated early because of increased mortality with D-sotalol. The group at greatest risk was those with a remote myocardial infarction and relatively good left ventricular function, the group that showed the lowest mortality when untreated. It is assumed that excessive prolongation of the action potential leading to pro-arrhythmia with D-sotalol, underlies the increased risk of death. However, there is little objective evidence in the SWORD trial to support this. Obviously D-sotalol should not be used in humans with a remote myocardial infarction and relatively good left ventricular function. D-Sotalol could still be considered for short-term hospital use in resistant arrhythmias and for longer-term use to prevent atrial fibrillation in those with remote myocardial infarction and poor left ventricular function. PMID- 11060767 TI - Eniluracil: an irreversible inhibitor of dihydropyrimidine dehydrogenase. AB - One of the most widely used drugs in cancer chemotherapy is 5-fluorouracil (5 FU). 5-FU is optimally delivered via continuous iv. infusion, which is both cumbersome and expensive. Prolonged oral dosing of 5-FU could mimic continuous infusion with less inconvenience and cost. However, oral administration of 5-FU has been hampered by incomplete and erratic bioavailability due to substantial variability in the activity of dihydropyrimidine dehydrogenase (DPD), the rate limiting enzyme in 5-FU catabolism. Eniluracil (ethynyluracil, GlaxoWellcome, USA), a uracil analogue, which irreversibly inhibits DPD, increases the oral bioavailability of 5-FU to 100%, facilitating uniform absorption and predictable toxicity. Cytotoxicity is enhanced one- to five-fold in cell lines treated with eniluracil plus 5-FU compared with 5-FU alone. Though eniluracil is neither toxic nor active as a single agent in animals, it improves the antitumour efficacy and therapeutic index of 5-FU. In Phase I trials, eniluracil markedly reduced the maximum tolerated dose of oral 5-FU, increased the half-life 20-fold and decreased the clearance 22-fold. DPD is completely inactivated within 1 h of eniluracil administration. Two dosing schedules have been evaluated in combination with oral 5-FU: a 5-day schedule every 28 days and a 28-day schedule every 35 days. The dose-limiting toxicity on the first schedule is myelosuppression with diarrhoea being dose-limiting on the 28-day schedule. Phase II trials employing the 28-day schedule have been completed in cancers of the colon, breast, liver and pancreas. Phase III trials in colorectal and pancreatic carcinoma have been completed and await analysis. Eniluracil is a promising drug, which permits reliable and safe administration of oral 5-FU and has the potential to overcome 5-FU resistance mediated by overexpression of DPD. PMID- 11060768 TI - Anti-inflammatory therapies in sepsis and septic shock. AB - Despite advances in supportive care, the morbidity and mortality rate resulting from sepsis and septic shock remain high (30 - 50%). A central hypothesis driving sepsis research in recent years is that this syndrome is the result of excessive inflammation. Therapies designed to inhibit the inflammatory response were first shown to be markedly beneficial in animal models of sepsis and then tested in numerous clinical trials involving thousands of patients. Three broad anti inflammatory strategies have been investigated. First, glucocorticoids in high doses administered at the onset of sepsis were studied. This approach proved unsuccessful. More recently, however, glucocorticoids in lower doses have been found to have a beneficial effect in patients with septic shock. Whether the mechanism of this treatment benefit is through inhibition of inflammation, or by counteracting a relative steroid refractoriness occurring during sepsis, remains unknown. The next focus of research were agents active against the endotoxin molecule. However, as with the experience with glucocorticoids, this approach lacked a consistent pattern of efficacy. It is unclear if this lack of efficacy is the result of endotoxin being a poor therapeutic target, or from testing agents which lacked the appropriate biological activity. Most recently, clinical trials in sepsis have focused on inhibiting specific host pro-inflammatory mediators (e.g., TNF, interleukins). While individual trials of inhibitors of these pro-inflammatory mediators failed to show a convincing benefit, pooling the results of these trials suggest that this approach has a marginal effect, supporting a role for excessive inflammation in sepsis. An unanswered question is reconcilling the very favourable effects obtained with anti-inflammatory treatments in animal models with the marginal results in humans. Further clinical and laboratory research is needed and may provide insight into more effective ways to use the anti-inflammatory agents already tested, or to investigate other potentially more effective anti-inflammatory agents in this syndrome. PMID- 11060769 TI - Antimicrobial resistance--why do we have it and what can we do about it? PMID- 11060770 TI - Anti-adhesive strategies in the prevention of infectious disease at mucosal surfaces. AB - Binding of microbial cell surface adhesins to host receptor molecules is a critical early step in microbial infection and pathogenesis. Anti-adhesive strategies aimed at blocking this interaction offer an attractive means of preventing infection at an early stage. The strategy should reduce the likelihood of resistant strains of microorganisms emerging, since those that do not bind will not be subjected to sustained selective pressure, as may occur with antibiotic therapy. Three classes of adhesion-blocking agent have been investigated, namely anti-adhesin antibodies, adhesin analogues and receptor analogues. The effectiveness of a number of these adhesion-blocking compounds has been demonstrated in human and animal models of infection. Direct application to the tooth surface of anti-adhesin monoclonal antibody, or a synthetic peptide adhesion epitope, prevented infection with the oral pathogen, Streptococcus mutans in humans. Intranasal administration of a soluble receptor analogue significantly reduced virus production and symptoms following experimental infection with rhinovirus. Similarly, all three types of anti-adhesion agent protected against a variety of infections at other mucosal surfaces in animal models. A common finding from these studies is the long duration of protection, which cannot be due to persistence of the anti-adhesion agent, but may be the result of competitive exclusion by members of the normal flora at specific mucosal surfaces. Development of these novel antimicrobial agents is particularly timely in view of the increasing concern over the spread of antibiotic resistance. PMID- 11060771 TI - Cationic antimicrobial peptides: towards clinical applications. AB - Cationic antimicrobial peptides are important components of the innate immune defences of all species of life. Variants of these natural molecules have a broad range of antibiotic, antifungal, antiviral and anti-endotoxic activity. Two of these cationic peptides have shown signs of efficacy in early clinical trials of oral mucositis and the sterilisation of central venous catheters, respectively and are currently proceeding through Phase III clinical trials. Thus, cationic antimicrobial peptides are currently being investigated as topical agents. In addition, the cationic protein rBPI 21 has recently completed Phase III clinical trials of parenteral use for meningococcaemia. PMID- 11060772 TI - Protegrins: new antibiotics of mammalian origin. AB - Protegrins and their derivatives are a new class of peptide antibiotics based on mammalian antimicrobial peptides. Their pharmacological properties include an unusually broad spectrum of antimicrobial activity against Gram-positive and Gram negative bacteria, fungi and some enveloped viruses. Preclinical and clinical studies of the lead compound, IB-367, developed for topical applications, show promise for the prevention of chemotherapy- and radiation-induced oral mucositis. PMID- 11060773 TI - Novel agents for the therapy of varicella-zoster virus infections. AB - Varicella-zoster virus (VZV), a member of the herpesvirus family, is responsible for both primary (varicella or chickenpox) as well as recurrent (zoster or shingles) infections. Acyclovir has been the mainstay for treating VZV infections in both immunocompetent and immunocompromised patients. Recently, newer anti-VZV drugs, i.e., valaciclovir (the oral prodrug form of acyclovir) and famciclovir (the oral prodrug form of penciclovir) have been developed and have enlarged the therapeutic options to treat VZV infections. Both acyclovir and penciclovir are dependent on the virus-encoded thymidine kinase (TK) for their intracellular activation. Although emergence of drug-resistant strains does not occur in immunocompetent patients, several reports have documented the isolation of drug resistant VZV strains following long-term acyclovir therapy in immunocompromised patients. Mutations at the level of the TK are responsible for development of resistance to drugs that depend on the viral TK for their phosphorylation (i.e., acyclovir and penciclovir). Foscarnet, a direct inhibitor of the viral DNA polymerase, which does not require activation by the viral TK, is the drug of choice for the treatment of TK-deficient VZV mutants emerging under acyclovir therapy. Recently, emergence of foscarnet-resistant strains has also been reported. Both TK-deficient strains and foscarnet-resistant mutants are sensitive to the acyclic nucleoside phosphonate cidofovir, CDV, HPMPC, (S)-1-(3-hydroxy-2 phosphonylmethoxypropyl)cytosine. This agent does not depend on the virus-encoded TK, but on cellular enzymes for its conversion to the diphosphoryl derivative, which then inhibits the viral DNA polymerase. PMID- 11060774 TI - Recent advances in diagnosis and therapy of human papillomaviruses. AB - Infection with human papillomavirus is extremely common throughout the world. Almost 50% of sexually active young women are infected with human papillomavirus and although most infections are transient, a subset has the potential to progress to invasive cancer. During the last 20 years, our understanding of the human papillomavirus life cycle and the role of human papillomavirus in human cancer has dramatically increased. Recent technological advances in human papillomavirus detection have provided the means to detect the presence of human papillomavirus with great sensitivity. In the context of patient care, there is still substantial debate regarding the optimal diagnostic and prognostic use of information derived from hybrid capture or polymerase chain reaction-based detection. The inventory of available treatment options is growing somewhat slowly. The most promising advances are being made in the clinical evaluation of candidates for prophylactic vaccination. This review is focused on the current status and future directions of prevention, diagnosis and therapy. PMID- 11060775 TI - Inhibitors of aminoacyl-tRNA synthetases as novel anti-infectives. AB - Resistance to existing antibiotics has emerged as a major problem in healthcare. Novel antibiotics for which bacteria have not yet acquired resistance need to be developed to combat drug-resistant pathogens. Aminoacyl-tRNA synthetases are leading targets for novel anti-infectives. The validation of aminoacyl-tRNA synthetases as drug targets for anti-infectives has been established in an animal system. Using several conceptually distinct approaches, new inhibitors of synthetases have been developed as drug prototypes. PMID- 11060776 TI - Developing G-quartet oligonucleotides as novel anti-HIV agents: focus on anti-HIV drug design. AB - Recently, a new class of oligonucleotides, forming G-quartet structures, has been developed as novel anti-HIV agents. Several critical structure-activity relationships between HIV-1 integrase and G-quartet oligonucleotides have been demonstrated. In addition the mechanism of the inhibition of HIV-1 integrase by G quartet oligonucleotides, such as T30695 and its derivatives, has been explored. This review summarises the preliminary studies of developing G-quartet oligonucleotides as novel anti-HIV agents in several aspects including structure activity relationship, stability-activity correlation, mechanism of HIV-1 integrase inhibition, substitution of phosphorothioates and targeting HIV-1 integrase in infected cells, which, hopefully, could help for developing a novel, efficient anti-HIV agent. PMID- 11060777 TI - Antifungals targeted to sphingolipid synthesis: focus on inositol phosphorylceramide synthase. AB - Currently available antifungal drugs for serious infections have essentially two molecular targets, 14alpha demethylase (azoles) and ergosterol (polyenes). The former is a fungistatic target, vulnerable to resistance development; the latter, while a fungicidal target, is not sufficiently different from the host to ensure high selectivity. Antifungals in clinical development have a third molecular target, beta-1,3-glucan synthase. Drugs aimed at totally new targets are required to increase our chemotherapeutic options and to forestall, alone or in combination chemotherapy, the emergence of drug resistance. Sphingolipids, essential membrane components in eukaryotic cells, but distinct in mammalian and fungal cells, present an attractive new target. Several natural product inhibitors of sphingolipid biosynthesis have been discovered in recent years, some of which act at a step unique to fungi and have potent and selective antifungal activity. PMID- 11060778 TI - New investigational antifungal agents for treating invasive fungal infections. AB - Systemic fungal infections have been recognised as a major cause of morbidity and mortality during the last two decades. There are only a few therapeutic options for these infections. Severe toxicity, such as impairment of renal function, limits the use of amphotericin B. Flucytosine is associated with side effects and drug resistance. Fluconazole and itraconazole are safer, though emergence of resistance and innate resistance in some fungal pathogens is a concern in their use. Therefore, there is a need for developing novel drugs and/or treatment strategies to combat these infections. In recent years, increased efforts by the pharmaceutical industry and academia have led to the discovery of new re engineered or reconsidered antifungal agents that are more efficacious, safer and have a broad spectrum of activity. Lipid formulations of polyene antifungal agents, amphotericin B and nystatin, have the advantage of improved therapeutic index. Activity against resistant fungi, high bioavailability, safety and longer half-life are the properties that encourage development of the newer triazoles (e.g., voriconazole, ravuconazole and posaconazole). Echinocandin-like lipopeptide antibiotics are among the antifungal agents with a novel mode of action. In addition to these lead investigational compounds, development of newer antifungal agents is underway. PMID- 11060779 TI - Artemisinin drugs: novel antimalarial agents. AB - Artemisinin and its derivatives, artesunate and artemether, represent a new class of antimicrobial drug with potent activity against Plasmodium falciparum. Although they show excellent efficacy in both severe and uncomplicated malaria, dosage regimens still need to be optimised and pharmacokinetic profiles defined. In the treatment of uncomplicated malaria, the artemisinin drugs should be used in combination with a long acting antimalarial to protect both drugs against the emergence of resistance. In the treatment of severe malaria, parenteral artemether is at least as effective as quinine and is simpler to use. The use of rectal preparations of artesunate and artemisinin at the rural health level will facilitate early initiation of the treatment of falciparum malaria and this may reduce the proportion of patients progressing to severe disease. All of the artemisinin drugs have comparable efficacy; the choice of derivative should be based upon availability, cost and quality of the preparation. Artemisinin, artesunate and artemether are well-tolerated in both adults and children, with no evidence to date of serious clinical toxicity. PMID- 11060780 TI - The therapeutic potential of phytoestrogens. AB - Phytoestrogens, such as the soya isoflavones genistein and daidzein, are currently being extensively investigated through both molecular, preclinical and clinical studies to determine their potential health benefits. Phytoestrogens may protect against chronic diseases such as hormone-dependent cancer (e.g., breast and prostate cancer), cardiovascular disease and osteoporosis. Investigations of phytoestrogen metabolism and bioavailability are also of great relevance. Conversion by gut microflora of daizein to its isoflavan metabolite equol, which is a more potent oestrogen and anti-oxidant, occurs only in some individuals (about 35% of subjects tested are equol excretors). This has considerable implications for daidzein bioavailability and also for cancer risk. Oxidative damage has been implicated in the development of heart disease and cancer and soya phytoestrogens have been reported to decrease plasma F(2)-isoprostane concentrations (biomarker for in vivo lipid peroxidation) and increase low density lipoprotein oxidation resistance. This anti-oxidant action of phytoestrogens could potentially contribute to their therapeutic efficacy. The findings from the current ongoing studies are all likely to contribute to determining the potential use of phytoestrogens as therapeutic agents. PMID- 11060781 TI - Squalene: potential chemopreventive agent. AB - Squalene is a triterpene that is an intermediate of the cholesterol biosynthesis pathway and it can be obtained from the diet. Olive oil contains 0.2-0.7% squalene. The average intake of squalene is 30 mg/day in the United States, however, when consumption of olive oil is high, the intake of squalene can reach 200-400 mg/day as observed in Mediterranean countries. The decreased risk for various cancers associated with high olive oil consumption may be due to the presence of squalene. Experimental studies have shown that squalene can effectively inhibit chemically-induced colon, lung and skin tumourigenesis in rodents. The protective effect is observed when squalene is given before and/or during carcinogen treatment. The mechanisms involved for the chemopreventive activity of squalene may include inhibition of Ras farnesylation, modulation of carcinogen activation and anti-oxidative activities. However, several factors must be taken into consideration when the evidence for the inhibition of carcinogenesis by squalene is examined, these include the effective dose used and the time of exposure. The information obtained is from animal bioassays and the long-term effects from consuming increased levels of squalene are not known. Although animal studies have enhanced our understanding of the possible action of squalene in decreasing carcinogenesis, one must apply caution in extrapolating the information obtained in animal studies to humans, because of possible species differences. In order to evaluate the overall implications of squalene to human cancer prevention, further studies are needed to fully identify its protective effects, as well as possible detrimental effects. PMID- 11060782 TI - Cyclin-dependent kinase inhibitors: novel anticancer agents. AB - In current models of cell cycle control, the transition between different cell cycle states is regulated at checkpoints. Transition through the cell-cycle is induced by a family of protein kinase holoenzymes, the cyclin-dependent kinases (CDKs) and their heterodimeric cyclin partner. Orderly progression through the cell-cycle involves co-ordinated activation of the CDKs, which in the presence of an associated CDK-activating kinase, phosphorylate target substrates including members of the 'pocket protein' family. This family includes the product of the retinoblastoma susceptibility gene (the pRb protein) and the related p107 and p130 proteins. Activity of these holoenzymes is regulated by post-translational modification. Phosphorylation of inhibitory sites on a conserved threonine residue within the activation segment is regulated by CDK7/cyclin H, referred to as CDK-activating kinase [1]. In addition, the cdc25 phosphatases activate the CDKs by dephosphorylating their inhibitory tyrosine and threonine phosphorylated residues [2,3]. Among the many roles for endogenous inhibitors (CDKIs), including members of the p21(CIP1/Waf1) family and the p16 family, one role is to regulate cyclin activity. Cellular neoplastic transformation is accompanied by loss of regulation of cell cycle checkpoints in conjunction with aberrant expression of CDKs and/or cyclins and the loss or mutation of the negative regulators (the CDKIs or the pocket protein pRb). One strategy to inhibit malignant cellular proliferation involves inhibiting CDK activity or enhancing function of the CDKI. Novel inhibitors of CDKs showing promise in the clinic include flavopiridol and UCN-01, which show early evidence of human tolerability in clinical trials. This review examines pertinent advances in the field of CDK inhibitors. PMID- 11060783 TI - Substance P antagonists: novel agents in the treatment of depression. AB - The field of neuropeptides has been expanding very rapidly in recent years. Apart from understanding their physiology and elucidating their functional role as putative neurotransmitters, research has focused on producing drugs that may treat a variety of illnesses in a novel way. Substance P antagonists occupy a central role in this area of intensive scientific activity. Substance P (SP), an undecapeptide, is abundant both in the periphery and in the CNS, where it is usually co-localised with one of the classical neurotransmitters, most commonly serotonin (5-HT). A role for SP is proposed in the regulation of pain, asthma, psoriasis, inflammatory bowel disease and, in the CNS, emesis, migraine, schizophrenia, depression and anxiety. A recently published positive study of MK 869, in depression, a novel SP antagonist has generated excitement amongst psychopharmacologists. It is the first time that a drug, not directly related to monoamine transmitters, has showed efficacy in depression. Although MK 869 has been suspended from further development, a host of other compounds, with similar action and better pharmacological profile, are currently under development. In this review, the pharmacology of central SP and its receptors are discussed, together with the exploration of the prospects and implications for future treatments of depression. PMID- 11060784 TI - Gatifloxacin: a new fluoroquinolone. AB - Gatifloxacin is a new 8-methoxy-fluoroquinoline antimicrobial agent. It has enhanced activity against Gram-positive and atypical agents, while retaining broad-spectrum antiGram-negative activity. For example, the MIC(90) values for respiratory tract pathogens are < or = 0.5 microg/ml for organisms such as Streptococcus pneumoniae (regardless of penicillin susceptibility), Haemophilus influenzae (beta-lactamase positive or negative), Moraxella catarrhalis (beta lactamase positive or negative), Legionella species, Mycoplasma pneumoniae, methicillin-sensitive Staphylococcus aureus, beta-haemolytic Streptococci (macrolide sensitive or resistant), Neisseria species, most Enterobacteriaceae, Neisseria gonorrhoeae, Neisseria meningitidis, Pasteurella species, Vibrio species and Yersinia enterocolitica. For methicillin-resistant S. aureus, ciprofloxacin-resistant S. aureus, Citrobacter freundii, Providencia species, Serratia species, Pseudomonas aeruginosa and other non-fermentative Gram-negative bacilli, the MIC(90) are elevated. Gatifloxacin is bactericidal and exhibits a post-antibiotic effect against Gram-positive and -negative bacteria. The standard dose is 400 mg once daily and is available in both oral and iv. formulation. Gatifloxacin appears to have a low propensity for the selection of resistant mutants. Clinical trial data supports the use of gatifloxacin for treatment of patients with respiratory tract, urinary tract, skin and soft tissue infections. The side effect profile for gatifloxacin is similar to that with other agents. PMID- 11060785 TI - Pharmacology and clinical experience with exemestane. AB - Since the introduction of the first generation aromatase inhibitor, aminoglutethimide, for breast cancer treatment 30 years ago, we now have at hand novel, potent and well-tolerated steroidal and non-steroidal compounds, allowing near complete inhibition of oestrogen synthesis. The third-generation aromatase inhibitor, or more accurately termed inactivator, exemestane, is a potent suppressor of oestrogen synthesis and is shown to be an effective antitumour agent in postmenopausal breast cancer patients. Exemestane has been shown to be effective in patients failing multiple endocrine regimens. A large randomised study has revealed that exemestane improves time-to-disease progression as well as overall survival compared with megestrol acetate as second-line therapy in patients failing tamoxifen. In current studies, exemestane is compared with tamoxifen as first-line therapy for metastatic disease. Sequential therapy with tamoxifen followed by exemestane is also being compared with tamoxifen monotherapy in the adjuvant setting. In addition, the drug may have potential for breast cancer prevention. PMID- 11060786 TI - Haemoglobin-based oxygen carriers. AB - Haemoglobin-based oxygen carriers are being developed for use in blood replacement therapies, either for perioperative haemodilution or for resuscitation from haemorrhagic blood loss. There is a high demand for these products because of risks associated with blood transfusions and pending worldwide blood shortages. Development of these products has required new technologies in protein engineering; since the haemoglobin is cell-free in solution, the molecule must be modified to be retained within blood circulation. Three classes of haemoglobin are under development: intramolecular cross-linked, intermolecular polymerised and surface conjugated with polyethylene glycol. Two products based on cross-linking chemistry have been discontinued because of serious adverse events and/or increased mortality rate in Phase III clinical trials. Three products based on polymerisation chemistry are in ongoing Phase III clinical trials. A new product based on surface conjugation is in preclinical evaluation. Although cross-linked and polymerised products have shown to be safe in preclinical and early Phase I/II clinical trials, they have had difficulty in proving efficacy. The primary adverse effect for the majority of cross-linked or polymerised products is a haemodynamic response, leading to increased vascular resistance to blood flow. The physiological mechanisms are still incompletely understood, so that safety and efficacy cannot be completely dissociated. New understandings on the mode of action of these products will help to define their utility and application. New products are under development, designed specifically to maximise blood flow and tissue perfusion and therefore, oxygenation. PMID- 11060787 TI - Therapeutic developments in peroxisome biogenesis disorders. AB - Clinically, peroxisome biogenesis disorders (PBDs) are a group of lethal diseases with a continuum of severity of clinical symptoms ranging from the most severe form, Zellweger syndrome, to the milder forms, infantile Refsum disease and rhizomelic chondrodysplasia punctata. PBDs are characterised by a number of biochemical abnormalities including impaired degradation of peroxide, very long chain fatty acids, pipecolic acid, phytanic acid and xenobiotics and impaired synthesis of plasmalogens, bile acids, cholesterol and docosahexaenoic acid. Treatment of PBD patients as a group is problematic since a number of patients, especially those with Zellweger syndrome, have significant neocortical alterations in the brain at birth so that full recovery would be impossible even with postnatal therapy. To date, treatment of PBD patients has generally involved only supportive care and symptomatic therapy. However, the fact that some of the milder PBD patients live into the second decade has prompted research into possible treatments for these patients. A number of experimental therapies have been evaluated to determine whether or not correction of biochemical abnormalities through dietary supplementation and/or modification is of clinical benefit to PBD patients. Another approach has been pharmacological induction of peroxisomes in PBD patients to improve overall peroxisomal biochemical function. Well known rodent peroxisomal proliferators were found not to induce human peroxisomes. Recently, our laboratory demonstrated that sodium 4-phenylbutyrate induces peroxisome proliferation and improves biochemical function (very long chain fatty acid beta-oxidation rates and very long chain fatty acid and plasmalogens levels) in fibroblast cell lines from patients with milder PBD phenotypes. Dietary supplementation and/or modification and pharmacological induction of peroxisomes as treatment strategies for PBD patients will be the subject of this review. PMID- 11060788 TI - Novel approaches to Behcet's disease. AB - Behcet's disease is a systemic inflammatory disorder. The patients have repeated exacerbations and remissions of the symptoms. This disease may produce a wide variety of symptoms. In mild cases, mucocutaneous lesions are only the symptoms during the whole clinical course, whereas ocular lesions, which occur in about 70% of the patients, can cause blindness. Involvement of the gastrointestinal tract, CNS and large vessels is less frequent, but sometimes life-threatening. Colchicine, NSAIDs, corticosteroids and immunosuppressants are employed for the treatment of Behcet's disease with therapies tailored to individual patients depending on clinical manifestations. Cyclosporin A is the most effective drug for ocular lesions at the present, but its neurotoxicity, which occurs in 20-30% of patients receiving cyclosporin A, restricts usage of the agent. Many patients are still suffering from a severe form of uveitis and serious neurological symptoms, which are resistant to any conventional therapies. New drugs have been investigated for Behcet's disease. IFN-alpha therapy has shown significant efficacy for common symptoms including ocular lesions without any serious adverse effects. Thalidomide and its analogues also appear to be applicable to this disease. Monoclonal antibody to TNF-alpha is now in clinical trials. These novel therapeutic approaches may provide much needed treatment options for patients with Behcet's disease. PMID- 11060789 TI - Sjogren's syndrome: current therapies remain inadequate for a common disease. AB - Sjogren's syndrome (SS) is a systematic autoimmune disease characterised by dysfunction of the lacrimal and salivary glands. This dryness leads to the symptoms of dry eyes and keratoconjunctivitis sicca, which is painful and may predispose patients to ocular infections. Also, SS patients develop dry mouth, which is uncomfortable and associated with progressive dental disease. SS is divided into secondary SS (where the dryness symptoms are associated with another well defined autoimmune disorder such as rheumatoid arthritis, systemic lupus erythematosus, or scleroderma) and primary SS (where the patients do not fulfil criteria for another well defined associated autoimmune disease). Primary SS has extra glandular organ involvement including lung (interstitial pneumonitis), renal (interstitial nephritis), peripheral and central nervous system manifestations, vasculitis of skin and other organs and increased frequency of lymphoma. This review will concentrate on primary SS. Therapies are divided into agents for topical replacement of deficient secretions (artificial tears, artificial salivas), stimulation of muscarinic M3 receptors (pilocarpine, cevimeline) to increase aqueous secretions, reduction of topical inflammation (topical cyclosporin or corticosteroids for the eye and fluorides or antibacterial varnishes for the mouth) and modification of the immune response in a manner similar to treatment of systemic lupus (antimalarial drugs, methotrexate, cyclophosphamide and perhaps newer agents such as leflunomide or TNF inhibitors). PMID- 11060790 TI - Immunoadsorption, current status and future developments. AB - The association of abnormalities in the cellular and humoral immune system with various autoimmune diseases provides the rationale for apheresis technologies. While plasmapheresis or plasma exchange is limited by its non-selective removal of all plasma components, modern apheresis techniques aim to provide more specific elimination according to clinical needs and avoid plasma product replacement. However, the commercialisation has not met the expectations in the early 80's and the number of patients treated by extracorporeal immunoadsorption remains small due to a lack of well-defined controlled trials and limited reimbursement. This review highlights the immunological and technical basis for extracorporeal immunoadsorption, as well as its current status in the treatment of immunologically-mediated diseases. PMID- 11060791 TI - Potential use of non-classical pathways for the transport of macromolecular drugs. AB - Since an increasing number of drug delivery strategies utilising proteins and peptides exhibiting 'non-classical' transport activities have been proposed, studies have begun to establish underlying functional relationships between different vectors. These attempts to find common factors have been hampered by a lack of biophysical data for the various potential protein and peptide transporters, as well as by the structural and functional diversity of the group as a whole. We describe the various types of vectors being considered for use and the preliminary therapeutic successes that have been achieved. Additionally, the various models that have been proposed for non-classical import and export are outlined and discussed in relation to therapeutic delivery. Possible future developments are also discussed. PMID- 11060792 TI - Animal models in xenotransplantation. AB - The severe shortage of donor organs has provided a strong impetus to push the investigation into the use of animal organs for humans. Xenotransplantation will not only benefit patients, but also represents a unique and potentially profitable business opportunity. However, there are many barriers to successful clinical xenotransplantation, including immunological barriers, physiological incompatibility, zoonosis and ethical concerns. This overview will focus on currently available animal models used in attempts to break through the immunological barriers to xenotransplantation. There are many advantages to using small animal, namely rodent, models in xenotransplantation research. For example, the use of the mouse model allows the use of knockout mice and careful dissection of rejection mechanisms at the molecular level. The following models can be used to study hyperacute rejection (HAR): guinea-pig-to-rat, mouse-to-rabbit, guinea pig-to-mouse, rat-to-presensitised mouse and rat-to-alpha-Gal knockout mouse. The hamster-to-rat, mouse-to-rat and rat-to-mouse models are commonly used to study acute vascular rejection. Large animal models are complex and expensive, but they are more relevant to clinical xenotransplantation. Based on experiments using transgenic pig-to-primate models, HAR can be overcome. However, acute vascular rejection remains a major barrier at the present time. A pig cartilage-to-monkey model has been developed to study chronic rejection. Other novel models such as pig venous segment-to-monkey model and rat-to-primate model may represent viable options to study immunological barriers following xenotransplantation. Like many other medical breakthroughs, animal research will continue to make enormous contributions towards the eventual success of xenotransplantation. PMID- 11060793 TI - Advances in skin gene therapy. AB - Specific anatomical and biological properties make the skin a very interesting target organ for gene therapy approaches. Different cell types of the epidermis, such as keratinocytes, melanocytes, or dendritic cells, can be genetically modified to treat a broad spectrum of diseases, including genetically inherited skin disorders, tumour diseases, metabolic disorders and infectious diseases. The easy accessibility of skin suggests that different methods for gene delivery can be pursued, depending on the desired application. The approach used to deliver DNA to the skin will influence not only the efficiency of DNA delivery, but also the level and duration of transgene expression. Furthermore, the desired biological effect will also influence the decision of which gene transfer method is the best choice. Among the current challenges of cutaneous gene therapy are: optimising the efficiency of direct in vivo gene delivery; targeting specific epidermal cells, including keratinocyte stem cells; achieving sustained gene expression and regulating gene expression in vivo. This review summarises recent advances in the field of skin gene therapy and evaluates possible strategies to overcome obstacles and achieve successful clinical applications of skin gene therapy. PMID- 11060794 TI - Cancer treatment with inhibitors of urokinase-type plasminogen activator and plasmin. AB - The urokinase-type plasminogen activator-plasmin system plays an important role in many normal physiological processes including clot lysis, wound healing, embryogenesis and tissue remodelling. It is also involved in the pathogenesis of human malignancy through its ability to mediate tumour cell growth, invasion and metastatic dissemination. Interfering with this system is an appealing approach for experimental therapy of malignancy for several reasons. This concept is supported by a wealth of preclinical data. Evidence exists suggesting a role for this system in several major human tumour types. Preliminary evidence suggests that agents which block this pathway are effective in therapeutic doses that are already defined and relatively non-toxic. This form of treatment is not likely to carry cross-resistance with other types of cancer therapy and should be applicable to both localised and advanced tumours. Since heterogeneity in responsiveness among various tumour types is expected, clinical effects in given tumours would provide a basis for interpreting mechanisms of tumour progression in vivo and for future development of drugs with improved efficacy. Inhibition of the urokinase-type plasminogen activator-plasmin system remains a promising, but largely untested, area of experimental cancer therapeutics. PMID- 11060795 TI - Progress in active-specific immunotherapy of brain malignancies. AB - Despite the significant advances in neurosurgical techniques and oncology treatment regimens, the prognosis of patients with brain malignancies remains dismal. Brain tumours remain as lethal in the beginning of this new millennium as they were 30 years ago. Among the promising treatment modalities being tested are various immunotherapeutic approaches. Development of cancer vaccines, also known as active-specific immunotherapy, for malignant brain tumours is summarised in this review. Understanding the mechanisms behind vaccinations and the initiation of immune response have helped the design and improvement of the efficacy of clinical vaccines. The emergence of the antigen-presenting properties of dendritic cells brings the cancer vaccine field into a new generation. Preclinical work on the use of dendritic cell-based vaccine for malignant brain tumours are encouraging. The move from these preliminary studies to the clinic is anticipated with high hope. PMID- 11060796 TI - The therapeutic potential of flavonoids. AB - Four most widely investigated flavonoids, flavopiridol, catechins, genistein and quercetin are reviewed in this article. Flavopiridol is a novel semisynthetic flavone analogue of rohitukine, a leading anticancer compound from an Indian tree. Flavopiridol inhibits most cyclin-dependent kinases and displays unique anticancer properties. It is the first cyclin-dependent kinase inhibitor to be tested in Phase II clinical trials. Catechin and its gallate are major ingredients in green tea and their anti-oxidant and cancer preventive effects have been widely investigated. A Phase I study of green tea extract GTE-TP91 has been conducted in adult patients with solid tumours. Similarly, genistein is a major ingredient in soybean and has been shown to prevent cancer and have antitumour, anti-oxidant and anti-inflammatory effects. Two antibody-genistein conjugates, B43-genistein and EGF-genistein, are currently in clinical development for the treatment of acute lymphoblastic leukaemia and breast cancer, respectively. Finally, most recent updates of quercetin are briefly described. PMID- 11060797 TI - Potential use of lipoxygenase inhibitors for cancer chemoprevention. AB - Increasing evidence suggests that lipoxygenase (LO)-catalysed metabolites have a profound influence on the development and progression of human cancers. Compared with normal tissues, significantly elevated levels of LO products have been found in breast tumours, colon cancers, lung, skin and prostate cancers, as well as in cells from patients with both acute and chronic leukaemias. LO-mediated products elicit diverse biological activities needed for neoplastic cell growth, influencing growth factor and transcription factor activation, oncogene induction, stimulation of tumour cell adhesion and regulation of apoptotic cell death. Agents that block LO catalytic activity may be effective in preventing cancer by interfering with signalling events needed for tumour growth. In the past ten years, pharmaceuticals agents that specifically inhibit the 5-LO metabolic pathway have been developed to treat inflammatory diseases such as asthma, arthritis and psoriasis. Some of these compounds possess anti-oxidant properties and may be effective in preventing cancer by blocking free radical induced genetic damage or by preventing the metabolic activation of carcinogens. Other compounds may work by negatively modulating DNA synthesis. Pharmacological profiles of potential chemopreventive agents are compiled from enzyme assays, in vitro testing (e.g., cell proliferation inhibition in human cancer cells) and in vivo animal carcinogenesis models (e.g., N-methyl-N-nitrosourea-induced rat mammary cancer, benzo(a)pyrene-induced lung tumours in strain A/J mice and hormone-induced prostate tumours in rats). In this way, compounds are identified for chemoprevention trials in human subjects. Based on currently available data, it is expected that the prevention of lung and prostate cancer will be initially studied in human trials of LO inhibitors. PMID- 11060798 TI - Focus on HDL: a new treatment paradigm for athero-thrombotic vascular disease. AB - Atherosclerotic vascular disease is the leading cause of mortality and morbidity in much of the Western world. Although advances in lifestyle and risk factor modification, pharmacotherapy, endovascular interventions and surgery have considerably improved clinical outcome, 40 - 50% of adverse cardiovascular events continue to occur despite current strategies. A number of new targets for therapeutic exploitation are currently being investigated that include, among others, apolipoprotein A-I, the major structural component of high density lipoprotein (HDL) particle. The strong negative relationship between HDL cholesterol levels and coronary heart disease in epidemiological studies, as well as data from experimental models suggest that HDL-based therapies could be an important new paradigm for prevention, treatment and reversal of atherosclerotic vascular disease. PMID- 11060799 TI - CTLA4-Ig: a novel immunosuppressive agent. AB - Activation of naive T-cells requires two signals: one is antigen-specific and based on T-cell receptor (TCR) recognition of a peptide-MHC complex and the second is antigen-nonspecific and delivered by specific T-cell receptors after ligation with their ligands (costimulatory molecules) expressed by antigen presenting cells (APCs). Engagement of the B7 family of molecules on APCs with their T-cell associated ligands, CD28 and CTLA-4 (CD152), provides a pivotal costimulatoty signal in T-cell activation. The lack of costimulation after engagement of the T-cell receptor by antigen, results in a state of antigen specific unresponsiveness, termed anergy. Manipulation of CD28/B7 pathway has therefore been envisioned as a potential strategy for achieving therapeutically useful immunosuppression or tolerance. CTLA4-Ig has been initially developed by Bristol-Myers Squibb as a competitive inhibitor of CD28/B7 pathway (BMS-188667). Thereafter, CTLA4-Ig was produced by Repligen and also in some individual laboratories. In various animal models, discussed in this paper, CTLA4-Ig has been shown to inhibit T-cell-dependent antibody responses, significantly prolong transplanted organ survival, induce long-term donor-specific tolerance in some models, slow progression of autoimmune disease and to have immunomodulatory function in several other immunological disease models. Recently, CTLA4-Ig has entered Phase I clinical trials for the treatment of psoriasis, a T-cell mediated skin disease and treatment of graft-versus-host disease in allogeneic bone marrow transplantation. Large clinical randomised trials on the use of CTLA4-Ig are missing, nevertheless, its immunosuppressive effects coupled with features such as specificity of interaction and low toxicity, make CTLA4-Ig a promising new therapeutic agent for induction of donor-specific immunological tolerance, the ultimate goal of clinical immunosuppression. PMID- 11060800 TI - Prinomastat, a hydroxamate-based matrix metalloproteinase inhibitor. A novel pharmacological approach for tissue remodelling-related diseases. AB - Prinomastat (formerly AG3340, Agouron Pharmaceuticals, Inc.) is a potent, selective oral inhibitor of matrix metalloproteinase-2, -9, -13 and -14. This peculiar selectivity should represent an advantage for prinomastat in terms of efficacy/tolerability. The drug has been shown to inhibit tumour growth and angiogenesis in a variety of preclinical models, including cancer of colon, breast, lung and intriguingly in melanoma and glioma models. Moreover, the combination of prinomastat and several chemotherapeutic agents was shown to induce additive effects. The drug is currently in Phase III clinical trials for patients with non-small cell lung cancer in combination with paclitaxel and carboplatin, as well as in advanced hormone refractory prostate cancer in combination with mitoxantrone. The most common side effects are musculoskeletal pain and stiffness. These side effects generally cease with treatment interruption. Finally, considering the pathophysiology of MMPs, Agouron is exploring the utility of prinomastat in ophthalmology and dermatology. PMID- 11060801 TI - Ongoing trials with matrix metalloproteinase inhibitors. AB - Excessive or poorly regulated matrix metalloproteinase (MMP) activity has been implicated as a pathogenic factor in a range of diseases where the extracellular matrix is degraded or remodelled. Synthetic, potent, low molecular weight MMP inhibitors (MMPIs) have been developed and, over the past five years, these agents have begun clinical testing in patients with cancer, rheumatoid arthritis, osteoarthritis and acute macular degeneration. The past year has seen a number of disappointments with the halting of clinical trials of Ro 32-3555 in patients with rheumatoid arthritis and of BAY 12-9566 in patients with cancer. There have, however, been some successes with perhaps the clearest indication of efficacy being seen in the results of a Phase III trial of marimastat in patients with advanced gastric cancer. Clinical trials are continuing with marimastat and other MMPIs, including prinomastat, solimastat, BMS 275291, metastat and neovastat. Results from these trials are expected in the next two years and it is likely that clinical trials with MMPIs will begin in patients with other diseases where MMPs are believed to be involved, such as restenosis, cerebral haemorrhage and multiple sclerosis. Future research is likely to focus on the identification of specific MMP targets in different diseases, both in order to improve efficacy and to reduce the musculoskeletal side effect profile that has characterised several of the first generation oral MMPIs. PMID- 11060802 TI - Drugs in development for social anxiety disorder: more to social anxiety than meets the SSRI. AB - Individuals with social phobia (SP) fear and avoid a wide variety of social and performance situations in which they are exposed to unfamiliar persons or to possible scrutiny by others. The lifetime prevalence of SP is estimated to be as high as 13%. It is frequently co-morbid with and usually precedes the onset of other psychiatric illnesses and is associated with significant occupational and social impairment, including academic and vocational underachievement. Fortunately, there are effective treatments for this common and debilitating condition. There is currently considerable evidence for the efficacy of pharmacotherapy and especially the monoamine oxidase inhibitors (MAOIs) and selective serotonin re-uptake inhibitors (SSRIs) in the treatment of this disorder. However, SSRIs are generally preferred as the first-line treatment of choice due to the advantages of SSRIs over MAOIs in terms of safety and tolerability. Despite encouraging results, current treatments most often produce partial symptomatic improvement, rather than high end-state functioning. While current first line treatments for social phobia target the serotonergic system, it is important to remember that different social fears are likely to have different developmental roots and may be based on quite different neurobiological systems. In this article we provide a review of current pharmacotherapeutic options for SP, current knowledge of the neurobiology of SP, and a review of new and promising directions in pharmacological research. It is increasingly clear that serotonin (5-HT) is unlikely to be the whole story in SP and that other brain chemical systems, especially the dopaminergic, noradrenaline-corticotropin releasing hormone and gamma-aminobutyric acid (GABA) dependent systems, most probably have an important role to play in a substantial percentage of cases. A number of new and novel agents, including the substance P antagonists, GABA agonists and CRF antagonists show considerable promise in the treatment of SP. However, in order to enhance the understanding of the neurobiology and treatment response of SP, we need to develop more sophisticated theory-driven typologies of SP. PMID- 11060803 TI - Therapeutic potential of NMDA receptor antagonists in the treatment of alcohol and substance use disorders. AB - Despite the fact that the use of alcohol, nicotine and other drugs is the major external factor contributing to mortality in industrialised countries, there are few medications available to treat alcohol and substance use disorders. In recent years, major advances have been made in the understanding of the neurobiological basis for these disorders and these advances should lead to the development of new pharmacotherapeutics. A substantial amount of the research suggests that N methyl-D-aspartate (NMDA) receptor neurotransmission contributes to mediating the behavioural effects of alcohol and other drugs of abuse. This research supports the therapeutic potential of NMDA receptor antagonists in alcohol and substance use disorders. In this paper the authors present their opinion on the goals and stages of pharmacological treatment of these complex psychiatric disorders. Available preclinical research using designs that model aspects of alcohol and substance use disorders is summarised, with an emphasis on research published in the last two years. In animal models, NMDA antagonists inhibit physical dependence and the reinforcing effects of a variety of abused substances. The ability of NMDA antagonists to inhibit tolerance to drug effects and contribute possible antidepressant and anxiolytic effects are also important from the perspective of drug development. This review summarises the relevant clinical laboratory and treatment data. Finally, it presents the status of the current development of NMDA receptor antagonists and discusses candidates with the greatest potential for clinical development. PMID- 11060804 TI - Involvement of cholecystokinin within craving for cocaine: role of cholecystokinin receptor ligands. AB - In the brain, cholecystokinin (CCK) has been described to act as a central neurotransmitter or neuromodulator involved in functions such as food consumption, stress and anxiety. Recently, the CCK system has been involved in drug dependence phenomena and proposed to be correlated to a putative state of 'drug preferring' phenotype within free choice tests. CCK exerts its action in the CNS through at least two different G-protein coupled high affinity receptors, CCK1 and CCK2. Various selective CCK receptor agonists and antagonists have been synthesised. In particular, L-364,718 has been demonstrated to be a potent and selective CCK1 receptor antagonist, whereas L-365,260 is a potent and selective CCK2 receptor antagonist. More recently, GV150013 has been reported to be a highly selective CCK2 receptor antagonist. This paper reviews the putative role of the CCK system within drug dependence phenomena. In particular, it analyses the relationship between central CCK activity and the exhibition of spontaneous preference for drugs of abuse, such as cocaine or alcohol. The potential therapeutic role for CCK receptor antagonists is also discussed. PMID- 11060805 TI - Muscarinic M(1) agonists in the treatment of Alzheimer's disease. AB - The treatment of Alzheimer's disease attempts to correct cholinergic deficiency in the brain. In addition to the established, but restricted, efficacy of acetylcholinesterase inhibitors, attempts are being made to develop agents which will stimulate muscarinic receptors directly. This approach is logical and was found efficacious in several animal models of the disease; however none of these agents succeeded in clinical studies. Several reasons might account for this failure, which are discussed, as well as the prospects for the future. PMID- 11060806 TI - Is there a role for potassium channel openers in neuronal ion channel disorders? AB - Malfunction in ion channels, due to mutations in genes encoding channel proteins or the presence of autoantibodies, are increasing being implicated in causing disease conditions, termed channelopathies. Dysfunction of potassium (K(+)) channels has been associated with the pathophysiology of a number of neurological, as well as peripheral, disorders (e.g., episodic ataxia, epilepsy, neuromyotonia, Parkinson's disease, congenital deafness, long QT syndrome). K(+) channels, which demonstrate a high degree of diversity and ubiquity, are fundamental in the control of membrane depolarisation and cell excitability. A common feature of K(+) channelopathies is a reduction or loss of membrane potential repolarisation. The identification of K(+) channel subtype specific openers will allow the recovery of the mechanism(s) responsible for counteraction of uncontrolled cellular depolarisation. Synthetic agents that demonstrate K(+) channel opening properties are available for a variety of K(+) channel subtypes (e.g., K(ATP), BK(Ca), GIRK and M-channel). This study reviews the realistic therapeutic potential that may be gained in a broad spectrum of clinical conditions by K(+) channel openers. K(+) channel openers would therefore identify dysfunctional K(+) channel as therapeutic targets for clinical benefit, in addition being able to modulate normally functioning K(+) channels to gain clinical management of pathophysiological events irrespective of the cause. PMID- 11060807 TI - Therapeutic potential of anti-inflammatory drugs in focal stroke. AB - The importance of cytokines, especially TNF-alpha and IL-1beta, are emphasised in the propagation and maintenance of the brain inflammatory response to injury. Much data supports the case that ischaemia and trauma elicit an inflammatory response in the injured brain. This inflammatory response consists of mediators (cytokines, chemokines and adhesion molecules) followed by cells (neutrophils early after the onset of brain injury and then a later monocyte infiltration). De novo upregulation of pro-inflammatory cytokines, chemokines and endothelial leukocyte adhesion molecules occurs soon after focal ischaemia and trauma, as well as at the time when the tissue injury is evolving. The significance of this brain inflammatory response and its contribution to brain injury is now becoming more understood. In this review, we discuss the role of TNF-alpha and IL-1beta in traumatic and ischaemic brain injury and associated inflammation and the co operative actions of chemokines and adhesion molecules in this process. We also address novel approaches to target cytokines and reduce the brain inflammatory response and thus brain injury, in stroke and neurotrauma. The mitogen-activated protein kinase (MAPK), p38, has been linked to inflammatory cytokine production and cell death following cellular stress. Stroke-induced p38 enzyme activation in the brain has been demonstrated and treatment with a second generation p38 MAPK inhibitor, SB-239063, provides a significant reduction in infarct size, neurological deficits and inflammatory cytokine expression produced by focal stroke. SB-239063 can also provide direct protection of cultured brain tissue to in vitro ischaemia. This robust SB-239063-induced neuroprotection emphasises a significant opportunity for targeting MAPK pathways in ischaemic stroke injury and also suggests that p38 inhibition should be evaluated for protective effects in other experimental models of nervous system injury and neurodegeneration. PMID- 11060808 TI - Novel therapeutic approaches to Guillain-Barre syndrome. AB - Guillain-Barre syndrome is an autoimmune disease which occurs throughout the world. Whilst the majority of patients can expect a reasonable recovery, about 10% die and 10% are left disabled with current therapy. The standard treatment is a five day course of iv. immunoglobulin, given at a dose of 0.4 g/kg/day, with plasma exchange as an equally efficacious alternative. Steroids are ineffective in Guillain-Barre syndrome. All new potential therapeutic agents need to be tested in addition to the standard agents available. Future potential therapies are suggested by the study of the animal model experimental autoimmune neuritis in the Lewis rat. Whilst in theory it is possible to target the different stages of the immune response, in practice not all of the steps at which experimental autoimmune neuritis can be prevented will be translatable to human Guillain-Barre syndrome. This is because Guillain-Barre syndrome probably presents after the immune reaction has been ongoing for some time and therefore early aspects of the immune response cannot be prevented. Many of the possible measures would have widespread immunosuppressive effects which would be unacceptable to patients. Interfering with the immune response by attempting to block antigen binding or inducing tolerance may not be practical, owing to the possibility of exacerbating disease. Once we have a more thorough understanding of the pathogenesis of Guillain-Barre syndrome, then immune-specific therapy for Guillain-Barre syndrome may become a possibility, rather than general immunosuppressive measures. Trials of beta-interferon and of a combination of steroid and i.v. immunoglobulin are underway. A trial of a second course of i.v. immunoglobulin is planned. PMID- 11060809 TI - Transplantation therapy for Parkinson's disease. AB - This review paper will provide an overview of the advent of neural transplantation therapy and the milestones achieved over the last 20 years for its use in treating Parkinson's disease. A discussion of technical factors that influence the outcome of neural transplantation is presented, with emphasis given on three sections dealing with immunosuppressants, alternative grafts and trophic factors which have recently been the focus of basic research and development of early phase clinical trials. Some views on the clinical assessment of transplanted Parkinson's disease patients are given at the end of the paper, with a synopsis highlighting the importance of basic research in advancing the potential clinical benefits of neural transplantation therapy in the treatment of Parkinson's disease. PMID- 11060810 TI - Neuroimmunophilin ligands: evaluation of their therapeutic potential for the treatment of neurological disorders. AB - Neuroimmunophilin ligands are a class of compounds that hold great promise for the treatment of nerve injuries and neurological disease. In contrast to neurotrophins (e.g., nerve growth factor), these compounds readily cross the blood-brain barrier, being orally effective in a variety of animal models of ischaemia, traumatic nerve injury and human neurodegenerative disorders. A further distinction is that neuroimmunophilin ligands act via unique receptors that are unrelated to the classical neurotrophic receptors (e.g., trk), making it unlikely that clinical trials will encounter the same difficulties found with the neurotrophins. Another advantage is that two neuroimmunophilin ligands (cyclosporin A and FK-506) have already been used in humans (as immunosuppressant drugs). Whereas both cyclosporin A and FK-506 demonstrate neuroprotective actions, only FK-506 and its derivatives have been clearly shown to exhibit significant neuroregenerative activity. Accordingly, the neuroprotective and neuroregenerative properties seem to arise via different mechanisms. Furthermore, the neuroregenerative property does not involve calcineurin inhibition (essential for immunosuppression). This is important since most of the limiting side effects produced by these drugs arise via calcineurin inhibition. A major breakthrough for the development of this class of compounds for the treatment of human neurological disorders was the ability to separate the neuroregenerative property of FK-506 from its immunosuppressant action via the development of non immunosuppressant (non-calcineurin inhibiting) derivatives. Further studies revealed that different receptor subtypes, or FK-506-binding proteins (FKBPs), mediate immunosuppression and nerve regeneration (FKBP-12 and FKBP-52, respectively, the latter being a component of steroid receptor complexes). Thus, steroid receptor chaperone proteins represent novel targets for future drug development of novel classes of compounds for the treatment of a variety of human neurological disorders, including traumatic injury (e.g., peripheral nerve and spinal cord), chemical exposure (e.g., vinca alkaloids, Taxol) and neurodegenerative disease (e.g. , diabetic neuropathy and Parkinson's disease). PMID- 11060811 TI - Novel approaches to the treatment of primary amyloidosis. AB - Primary (AL, amyloid light-chain) amyloidosis is a plasma cell disorder in which deposits of amyloid light-chain protein cause progressive organ failure. It is important to recognise that amyloidosis is a dynamic process and chemotherapy induced reduction of the activity of the plasma cell clone reduces the supply of the amyloid precursor protein and can result in a major regression of the deposits. The most common target organ is the kidney and renal amyloidosis manifests as proteinuria or nephrotic syndrome. Proteinuria is seen in three quarters of patients. Amyloid related nephrotic syndrome and renal failure are potentially reversible. Fatigue, congestive heart failure, hepatomegaly, peripheral neuropathy, orthostatic hypotension, carpal tunnel syndrome and macroglossia are other common features. The median survival is one to two years. Conventional-dose melphalan as standard treatment can prolong the median duration of survival by about ten months, but the clinical response rates with improvement of impaired organ function are low. Up-front high-dose chemotherapy with autologous peripheral blood stem cell transplantation is much more effective and can result in a major improvement in the clinical condition of patients. However, the toxicity related to this treatment can be relevant due to impaired organ function. Conventional-dose chemotherapy consisting of vincristine, doxorubicin and dexamethasone or high-dose dexamethasone or interferon-alpha are other possible approaches to treatment. The improvement of patient condition with an effective conventional-dose chemotherapy may increase the tolerability of high dose chemotherapy and reduce transplantation related problems. PMID- 11060812 TI - Inhibitors of bacterial two-component signalling systems. AB - Bacterial two-component regulatory systems (TCS) play a pivotal role in the process of infection. These signal transduction systems enable bacterial pathogens to mount an adaptive response and cope with diverse environmental stresses, including nutrient deprivation, antibiotic onslaught and phagocytosis. Interest in these systems as novel bacterial targets has been rekindled by the recent discovery of several essential systems in important Gram-positive and Gram negative pathogens. Several series of TCS inhibitors derived from broad screening approaches have been reported in the literature, however, most appear to suffer from poor selectivity, excessive protein binding and/or limited bioavailability. Consequently, pharmaceutical chemists have turned to alternate strategies, such as the design of substrate-based inhibitors, the generation of combinatorial libraries and the isolation of natural products, to identify inhibitors with more desirable properties. Recent structural studies of the histidine protein kinase and response regulator proteins that constitute TCS may provide a foundation for a structure-based design approach to TCS inhibitors. PMID- 11060813 TI - Fibrin sealant in wound repair: a systematic survey of the literature. AB - Fibrin sealants have recently been approved for clinical use in the US by the FDA and have been available for clinical use in Europe for years. The indication for use in the US is haemostasis. Nevertheless, both commercial and non-commercial fibrin sealant preparations are also for wound healing and for prevention of abdominal adhesions in the US and Europe. To the non-cognoscenti of fibrin sealants, their use to promote wound repair and to prevent abdominal adhesions appears contradictory since an agent that promotes connective tissue repair might be expected to promote abdominal adhesion rather than prevent them. In this systematic survey of the animal and clinical data evidence is presented that supports both off-label uses. However there is much inconsistency in the data secondary to the use of various fibrin sealant preparations, different animal models and clinical situations and different application techniques. It is clear from this survey that standard preparation and application of fibrin sealant for a particular surgical setting are needed to resolve the many apparent discrepancies in the literature. A corollary to this is the likelihood that different fibrin sealant preparations may be preferred for different clinical situations. PMID- 11060814 TI - Review of the acetylcholinesterase inhibitor galanthamine. AB - Galanthamine (or galantamine, Reminyl) is a tertiary alkaloid acetylcholinesterase inhibitor (AChEI) which has been approved in several countries for the symptomatic treatment of senile dementia of the Alzheimer's type. Derived from bulbs of the common snowdrop and several Amaryllidaceae plants, (-)-galanthamine (GAL) has long been used in anaesthetics to reverse neuromuscular paralysis induced by turbocurarine-like muscle relaxants and more recently, has been shown to attenuate drug- and lesion-induced cognitive deficits in animal models of learning and memory. GAL directly inhibits acetylcholinesterase activity, while demonstrating much weaker activity on butyrylcholinesterase (BuChE). GAL also stimulates pre- and postsynaptic nicotinic receptors, although the clinical significance of this finding is yet unclear. Numerous variants and analogues of GAL have also been developed, with varying potency in inhibiting AChE activity. GAL is readily absorbed after oral administration, with a t(max) of 52 min and a plasma elimination t(1/2) of 5.7 h. The efficacy of GAL administered to Alzheimer's disease (AD) patients has been well demonstrated by large-scale clinical trials. Typical of AChEIs, the most common adverse events associated with GAL are nausea and vomiting. In conclusion, evidence to date suggests galanthamine to be similar to other AChEIs in improving cognitive function in AD patients. PMID- 11060815 TI - An evaluation of intrathecal ziconotide for the treatment of chronic pain. AB - Ziconotide, the synthetic form of cone snail peptide pi-conotoxin MVIIA, is a neurone-specific N-type calcium channel blocker with an analgesic and neuroprotective effect. Intrathecal ziconotide has been recommended for approval by the FDA for the management of chronic pain. Spinally administered ziconotide produces analgesia by blocking neurotransmitter release from primary nociceptive afferents and prevents the propagation of pain signals to the brain. It has an advantage over intrathecal morphine in that there is no development of tolerance after prolonged use. Systemic toxicity is considerably reduced by administration of smaller doses intrathecally and selective delivery to the site of action in the nervous system. Nevertheless, there are neurological adverse effects due to delay in clearance of ziconotide from the neural tissues. Overall, ziconotide has a favourable risk/benefit ratio with advantages over several currently available intrathecal therapies for pain. PMID- 11060816 TI - The association of infection and coronary artery disease: an update. AB - Numerous studies have reported an association of coronary atherosclerosis and restenosis with certain bacterial and viral infections. This article reviews the pathophysiology of atherosclerosis, the role of infectious agents (cytomegalovirus, Chlamydia pneumoniae and Helicobacter pylori) in atherogenesis and studies supporting the potential beneficial effects of antibiotics or antiviral agents in the management of atherosclerotic disease. The interactions of cytomegalovirus and the arterial wall have been extensively studied. However, despite the successful preliminary therapeutic trials with the use of macrolides in augmenting possible C. pneumoniae-induced cardiovascular events, the exact mechanisms of how C. pneumoniae enters the arterial wall remains unknown at this point. For H. pylori, regardless of the large number of studies performed to assess the association between H. pylori and coronary artery disease, no definitive conclusion could be made at this time, due to contradictory results. Before one can widely adopt the use of antibiotics or antiviral agents as treatment for atherosclerosis, further studies must be designed to address some important issues. In vivo animal models need to be established to further examine the various hypotheses regarding the interaction of infectious agents and atherosclerosis and restenosis. Large-scale prospective cohort studies should be designed to relate evidence of infection to future risk of cardiovascular diseases. Confounding variables, such as other cardiovascular risk factors and socio-economic status, should be controlled in order to strengthen the association. Further interventional studies are also required to establish the best antibiotic or antiviral regimen to maximise efficacy and minimise side effects. PMID- 11060817 TI - Adenosine therapy: a new approach to chronic heart failure. AB - Both the prevention and attenuation of chronic heart failure (CHF) are important issues for cardiologists. There are three different strategies to prevent patients from deleterious sequels. The first strategy is to remove the causes of CHF if possible; the second is to attenuate the events that may lead to CHF, such as myocardial ischaemia and reperfusion injury, cardiomyopathy and myocarditis, cardiac hypertrophy and ventricular remodelling; the third is to prevent or attenuate the progression of CHF. Adenosine has a number of actions which merit it as a possible cardioprotective and therapeutic agent for CHF. Firstly, adenosine induces collateral circulation via inducing growth factors and triggering ischaemic preconditioning, both of which induce ischaemic tolerance in advance. Adenosine is also known to reduce the release of noradrenaline, production of endothelin and attenuate the activation of renin-angiotensin system all of which are believed to cause cardiac hypertrophy and remodelling. Secondly, exogenous adenosine is known to reduce the severity of ischaemia and reperfusion injury. Thirdly, adenosine is reported to counteract neurohumoral factors, i.e., cytokine systems, known to be related to the pathophysiology of CHF. Recently, we revealed that adenosine metabolism is changed in patients with CHF and increases in adenosine levels may aid to reduce the severity of CHF. Thus, there are many potential mechanisms for cardioprotection attributable to adenosine and we postulate the use of adenosine therapy will be beneficial in patients with CHF. PMID- 11060818 TI - Therapeutic potential of H(3)-receptor agonists in myocardial infarction. AB - Sympathetic over-activity accompanied by excessive noradrenaline (NA) release within the heart is a recognised cause of dysfunction in myocardial ischaemia. Myocardial infarction is often accompanied by arrhythmias with high morbidity and mortality. Indeed, NA enhances intracellular Ca(2+) by increasing its influx through voltage-dependent channels, mobilising it from intracellular stores and favouring its inward transport by Na(+)/Ca(2+) exchange. Ca(2+) overload eventually results in dysrhythmia and uncoordinated myocyte contraction. Moreover, NA increases metabolic demand. In concert with other contributing factors, this will aggravate the primary ischaemia and initiate a vicious cycle that can culminate in myocardial damage and heart failure. Therefore, reduction of NA release from cardiac sympathetic nerves is an important protective measure. Adrenergic nerves possess inhibitory receptors, such as alpha(2)-adrenoceptors, adenosine A(1)-receptors and histamine H(3)-receptors (H(3)R). In myocardial infarction, NA is released by both exocytotic (Ca(2+)-dependent) and carrier mediated (Na(+)/H(+) exchange-dependent) mechanisms, associated with short-term and protracted ischaemia, respectively. Unlike alpha(2)-adrenoceptor agonists that reduce NA exocytosis, but enhance carrier-mediated NA release, H(3)R agonists inhibit both exocytotic and carrier-mediated NA release. Moreover, unlike adenosine A(1)-receptor agonists, H(3)R agonists do not depress sinoatrial and atrioventricular nodes, nor cause bronchoconstriction. Therefore, stimulation of H(3)R on cardiac sympathetic nerve endings is an important new way to protect the heart from the consequences of ischaemia and infarction. Although H(3)R agonists alleviate reperfusion arrhythmias in isolated hearts by reducing NA release, this protective action needs to be demonstrated in classical in vivo models of occlusion/reperfusion. Regardless, H(3)R agonists offer the promise of a novel strategy in the treatment of myocardial ischaemia and infarction. PMID- 11060819 TI - Therapeutic developments in sudden cardiac death. AB - Sudden cardiac death is characterised by the unexpected death of a patient who has been clinically stable. It is frequently due to the development of ventricular tachyarrhythmias. With appropriate treatment, patients can be appropriately resuscitated. Clinically, it is essential to develop treatment strategies to prevent such an episode, as most patients do not survive out-of hospital cardiac arrest. beta-Blockers are an effective pharmacological therapy in patients following myocardial infarction and in those with congestive heart failure. They may also be effective in other types of heart disease. Anti arrhythmic agents are not useful as prophylactic drug therapy for reducing mortality in patients at risk for sudden cardiac death. Amiodarone is a notable exception, which may have some benefit, particularly in some subgroups. The implantable cardioverter-defibrillator has emerged as the most effective therapy for preventing sudden cardiac death in high-risk patients. Further work is required to enhance the characterisation of high-risk patients. Genetic analyses in patients with cardiovascular disorders may also identify new approaches to the prevention of sudden cardiac death. PMID- 11060820 TI - Novel approaches for the prevention of restenosis. AB - Restenosis, the re-narrowing of the lumen of the coronary artery, in the months following a successful percutaneous balloon angioplasty or stenting, remains the main limitation to percutaneous coronary revascularisation. Serial intravascular ultrasound studies have shown that restenosis after conventional balloon angioplasty represents a complex interplay between elastic recoil, smooth muscle proliferation and vascular remodelling, while restenosis after stent deployment is due almost entirely to smooth muscle hyperplasia and matrix proliferation. Despite intensive investigation in animal models and in clinical trials, most pharmacological agents have been found to be ineffective in preventing restenosis after percutaneous balloon angioplasty or stenting. Although studies frequently report success in the suppression of neointimal proliferation in animal models of balloon vascular injury, few of them have been successful in clinical trials. Lately, the advent of endovascular radiation, new antiproliferative agents, recombinant DNA, growth factor regulators and novel local drug delivery systems have shown promising results. In the past five years, intracoronary radiation with gamma- and beta-emitting sources has been evaluated intensively with very encouraging results. This is the first potent non-pharmacological approach that has been successful in a large number of patients in controlling excessive tissue proliferation. It is very likely that a combination of stents and pharmacological and/or non-pharmacological inhibition of neointimal hyperplasia will likely result in further reductions in the incidence if restenosis. The continued attractiveness of percutaneous coronary revascularisation, as an alternative to medical treatment or bypass surgery for patients with coronary artery disease, will depend upon our ability to control the restenotic process. Due to the vast literature on the subject, this review will focus mainly on clinical trials that show the most promise and will highlight those that warrant further investigation. PMID- 11060821 TI - Novel approaches to the treatment of acute renal failure. AB - Acute renal failure (ARF) occurs frequently in hospitalised patients and is associated with significant morbidity and mortality. Many therapeutic strategies have been undertaken both to prevent acute renal injury and, once ARF occurs, to improve renal function and reduce mortality. Among the available pharmacological options, no specific therapy has been shown to alter the course of ARF. This article reviews the efficacy of several strategies in experimental renal disease and raises the possibility that similar interventions might be available to the clinician in the near future for the prevention and management of ARF. The prospect of these novel strategies together with the ever-increasing understanding of the complex pathophysiology of ARF, offers the promise of effective and more physiological therapeutic interventions in this new millennium. PMID- 11060822 TI - Renal protective potential of antihypertensive drugs. AB - The objective of this review is to discuss recent experimental and clinical data concerning the effectiveness of antihypertensive drugs in preventing or delaying renal changes caused by diabetes mellitus and hypertension and to examine possible future developments. A brief description of the mechanisms involved in the development of renal failure in diabetes and hypertension is included. Evidence is presented to show that in addition to renoprotection offered by reduction in arterial pressure, some antihypertensive agents may give more nephroprotection. This added renoprotective potential of antihypertensive agents, which are either already in use or are being developed, is discussed. The nephroprotective action of conventional antihypertensive drugs, such as beta blockers, calcium antagonists and angiotensin-converting enzyme inhibitors is briefly reviewed. It is noted that several studies indicate that angiotensin converting enzyme inhibitors may be more effective in preventing or retarding renal failure than other conventional drugs. The renoprotective potential of newly developed agents, such as angiotensin II Type 1 receptor antagonists, vasopeptidase inhibitors and endothelin receptor antagonists is also examined. Emphasis is placed on a possible superior renoprotective effect of combination therapy over monotherapy. PMID- 11060823 TI - Therapeutic potential of ACE inhibitors for the treatment of hypertension in Type 2 diabetes. AB - Type 2 diabetes mellitus is associated with hypertension. If untreated, hypertension has a major impact on the clinical course of Type 2 diabetes and its vascular complications. In this review, we discuss rationale for the use of ACE inhibitors (ACEI) in hypertensive Type 2 diabetic patients and compare those theoretical assumptions with results of recent major clinical trials. Furthermore, possible directions for future clinical and experimental research are outlined. The RAS and its effector angiotensin II are important players in a number of cardiovascular and renal disorders. Recent evidence suggests that RAS and factors functionally linked to RAS are activated in Type 2 diabetes. Therefore, there is a theoretical basis for the use of ACEI in the treatment of hypertension in diabetic patients. Some recent studies reported superior outcome in patients treated with ACEI-based antihypertensive regimens compared with non ACEI based treatments in reducing the risk of macrovascular disease (CAPPP, FACET, ABCD) or both micro- and macrovascular complications in Type 2 diabetes (HOPE). However, at least two of the large prospective studies discussed in this review (UKPDS 38, HOT), supported by results from previously published SHEP study, have recently suggested that the degree of reduction of blood pressure, rather than the choice of a particular class of antihypertensive agent, is associated with decreased risk of cardiovascular events. Studies focusing on renal end-points suggest that ACEI have a superior antiproteinuric effect than the other agents. However, whether ACEI are more nephroprotective, as assessed by the rate of decline in renal function, still remains to be elucidated. Despite promising results from recent trials, large numbers of patients progress despite ACEI treatment. Incomplete inhibition of the RAS may underlie this phenomenon. Treatment strategies that could enhance the degree of RAS inhibition represent one possible direction for clinical research in the near future. However, it is unlikely that the course of such a complex syndrome as Type 2 diabetes could be dramatically changed by just one class of antihypertensive agents. This goal is more likely to be achieved by multifactorial intervention, reflecting the complexity of metabolic syndrome. ACEI should be viewed as an important, but not the only, part of this complex approach. PMID- 11060824 TI - Novel lipid-regulating drugs. AB - This review describes the major developments in lipid-regulating drugs during the past two years. Most of the novel compounds that have been launched or were in their final stages of development during this period are aimed primarily at lowering low-density lipoprotein (LDL)-cholesterol. These include bile acid sequestrants, HMG-CoA reductase inhibitors and a cholesterol absorption inhibitor. In addition, there have been preclinical reports suggesting the potential usefulness of orally bioavailable inhibitors of cholesterol ester transfer protein in plasma and of acylcoA:cholesterol acyltransferase in monocyte macrophages. The recent discovery of the roles of the scavenger receptor class B Type 1 and ATP-binding cassette transporter 1 in high-density lipoprotein (HDL) cholesterol transport may shift the trend of future developments away from compounds that lower LDL to those aimed at promoting HDL turnover. PMID- 11060825 TI - Therapeutic potential of vitamin E in heart disease. AB - Low density lipoprotein (LDL) oxidation is considered an important step in the atherogenic process. Oxidatively modified particles induce the expression of adhesion molecules, stimulate the production of inflammatory cytokines and impair endothelial function. The measurement of oxidised LDLs in vivo is very difficult, therefore most investigators rely on the measurement of in vitro oxidability of these particles to evaluate their deleterious effects. Supplementation with water and lipid soluble anti-oxidant vitamins, especially vitamin C and E, significantly increase the resistance to LDL oxidation. Vitamin E supplementation also improves endothelium-dependent vasodilation in hypercholesterolaemic and subjects who smoke cigarettes. Epidemiological studies have not consistently demonstrated a protective effect of vitamin E consumption as food or supplements on coronary events or stroke. Likewise, only one of five large prospective trials has shown a beneficial effect of vitamin E supplementation on cardiovascular events or mortality. One report showed that supplemented haemodialysed patients had a lower incidence of cardiovascular events. Thus, presently, there is not enough evidence to widely recommend the use of vitamin E supplements for vascular protection. PMID- 11060826 TI - Therapeutic potential of total homocysteine-lowering drugs on cardiovascular disease. AB - An elevated total homocysteine (tHcy) plasma concentration is associated with increased morbidity and mortality due to cardiovascular disease in the general population and in patients with impaired renal function. The prevalence of hyperhomocysteinaemia (plasma levels above 15 micromol/l) in the general population is less than 5% and can be as high as 50% in patients with vascular disease. In patients with renal insufficiency, elevated tHcy plasma levels are detected in 50 - 100% of the patients. Total homocysteine plasma levels can be lowered or normalised by folic acid and/or vitamin B(6) and vitamin B(12) supplementation. In patients with advanced chronic renal insufficiency or end stage renal disease, hyperhomocysteinaemia is partially resistant to folic acid or vitamin therapy. However, higher tHcy plasma levels may also reflect tissue damage and the increase in Hcy after an acute incident such as stroke or myocardial infarction may be necessary for tissue repair mechanisms. This implies, that lowering tHcy may even be harmful to some patients. Currently, prospective studies are underway to clarify whether folate supplementation, with or without additional other vitamins, improves cardiovascular disease morbidity and mortality in the general population, as well as in renal failure patients. While population-wide screening for and treatment of hyperhomocysteinaemia is generally not recommended, treatment of high risk patients may be considered. PMID- 11060827 TI - NK-104: a novel synthetic HMG-CoA reductase inhibitor. AB - An elevated level of low-density lipoprotein (LDL)-cholesterol has been recognised as the most important risk factor for coronary artery disease (CAD). Development of the inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) ('statins'), a rate-limiting key enzyme of cholesterol synthesis pathway, has revolutionised the cholesterol-lowering therapy. In the last decade, effective primary and secondary preventive measures have been established in several statin trials to prevent future events of CAD by lowering LDL-cholesterol levels. These results supported the 'lower is better' hypothesis in the relationship between LDL-cholesterol levels and CAD. NK-104 (pitavastatin, previously named as itavastatin or nisvastatin, Kowa Company Ltd., Tokyo) has recently been developed as a new chemically synthesised and powerful statin. On the basis of reported data, the potency of NK-104 is dose-dependent and appears to be equivalent to that of atorvastatin. This new statin is safe and well-tolerated in the treatment of patients with hypercholesterolaemia. The cytochrome P450 system only slightly modifies NK-104, which suggests the clinical advantage of this agent, because the prevalence of clinically significant interactions with a number of other commonly used drugs can be considered to be extremely low. NK-104 can provide a new and potentially superior therapeutic agent when compared with currently available other statins. Randomised controlled clinical trials to assess the long-term effects of this new statin on CAD would be required. PMID- 11060828 TI - Colesevelam hydrochloride: a non-absorbed, polymeric cholesterol-lowering agent. AB - Colesevelam hydrochloride (formerly known as Cholestagel((R)) and re-named WelCholtrade mark, GelTex Pharmaceuticals, Inc. and Sankyo Parke-Davis) is a new, polymeric, high potency, water-absorbing hydrogel. It has been shown to be a safe and effective cholesterol-lowering agent with a non-systemic mechanism of action, good tolerability and minimal side effects. To date, the lipid-lowering activity of colesevelam has been evaluated in approximately 1400 subjects. Colesevelam reduces low density lipoprotein (LDL)-cholesterol levels, in a dose-dependent manner, by as much as 20% (median) in patients with hypercholesterolaemia. Dosing regimen evaluations indicate that colesevelam is effective at both once per day and twice daily dosing and that concurrent administration of colesevelam with hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins), specifically lovastatin, does not alter the absorption of the statin. Combination therapy with HMG-CoA reductase inhibitors, including lovastatin, simvastatin and atorvastatin, produces an additional reduction (8 - 16%) in LDL-cholesterol levels above that seen with the statin alone. The overall incidence of adverse effects with colesevelam alone and in combination with statins is comparable with that seen with placebo. Colesevelam lacks the constipating effect seen with typical bile acid sequestrants, a trait that would be expected to improve compliance with lipid-lowering therapy. Colesevelam, recently approved by the US FDA, represents a valuable non-absorbed alternative in the armamentarium against hypercholesterolaemia, both for monotherapy and combination therapy, as an adjunct to diet and exercise. PMID- 11060829 TI - Lotrafiban: an oral platelet glycoprotein IIb/IIIa blocker. AB - Platelets play a major role in thrombus formation, as well as in the pathogenesis of atherothrombosis. Inhibition of platelet function is now emphasised more than ever for prevention and treatment of almost all vascular diseases, since thrombosis is established as the key pathogenic event causing acute ischaemic coronary and cerebrovascular syndromes. Although acetylsalicylic acid (aspirin) has been shown to reduce the incidence of myocardial infarction and stroke, its effect is weak and more effective antithrombotic agents are required to manage patients at high-risk for recurrent vascular events. Platelet glycoprotein IIb/IIIa receptor (GPIIb/IIIa) blockade represents a significant advance in interventional cardiology and treatment of acute ischaemic syndromes. The past several years have seen the introduction of many platelet GPIIb/IIIa blockers into the clinical arena targeting the unique platelet GPIIb/IIIa receptor for the adhesive proteins, fibrinogen and von Willebrand Factor. Platelet GPIIb/IIIa blockers administered intravenously have proven efficacious in mitigating arterial thrombosis in acute coronary syndromes (unstable angina and non-ST elevation myocardial infarction) and percutaneous coronary interventions (PCI) such as balloon dilatation and stent implantation. Currently, orally-active platelet GPIIb/IIIa blockers are being developed to provide additional benefits for primary and secondary prevention of thrombosis as chronic treatment, especially in high-risk patients. Lotrafiban (SmithKline Beecham) is a member of the latest generation of orally-active platelet GPIIb/IIIa blockers undergoing Phase III clinical trials to test the relative effectiveness versus other oral platelet inhibitors for ischaemic conditions including unstable angina, restenosis after PCI and stroke. Lotrafiban is converted from an esterified prodrug by plasma and liver esterases to a peptidomimetic of the arginine-glycine aspartic acid amino acid sequence. This sequence itself mimics the binding site of fibrinogen and von Willebrand Factor to the platelet GPIIb/IIIa receptor. Preliminary results of the clinical trial APLAUD (antiplatelet useful dose) show that lotrafiban is clinically safe and well-tolerated in patients with recent myocardial infarction, unstable angina, transient ischaemic attack (TIA), or stroke when added to aspirin therapy. With lotrafiban, a worldwide large-scale Phase III clinical trial BRAVO (blockage of the GPIIb/IIIa receptor to avoid vascular occlusion) is currently underway. In general, GPIIb/IIIa blockade seems clinically very promising. A number of unresolved issues, however, remain to be elucidated. PMID- 11060830 TI - Pharmacology and clinical trial results of lanoteplase in acute myocardial infarction. AB - New bolus fibrinolytic agents derived from the recombinant human tissue plasminogen activator (t-PA) have emerged as a new means of dissolution of the occlusive thrombosis associated with acute myocardial infarction. Lanoteplase is a fibrinolytic drug derived from t-PA by deleting its fibronectin finger-like and epidermal growth factor domains and mutating Asn(117) to Gln(117). Lanoteplase has a reduced plasma clearance and a prolonged half-life such that it can be administered as a single bolus. In the InTIME I trial, patency (TIMI grade 2 or 3 flow) with the 120 KU/kg dose was higher compared with front-loaded t-PA. The InTIME II trial demonstrated that lanoteplase was as effective as alteplase with regard to mortality. However, the rate of intracranial haemorrhage was significantly higher in lanoteplase-treated patients and further development of this compound has been halted. PMID- 11060831 TI - Dofetilide: a class III anti-arrhythmic drug for the treatment of atrial fibrillation. AB - Dofetilide is a class III anti-arrhythmic drug that has been approved for the treatment of atrial fibrillation. Two clinical studies, which enrolled 996 patients, demonstrated pharmacological conversion to sinus rhythm to occur in 30% of patients. Following pharmacological or electrical conversion, median time to relapse exceeded one year. Two large clinical studies that enrolled 3028 patients have been performed in high-risk patients with severe heart failure and large myocardial infarctions. The outcomes of these studies were neutral with respect to survival and demonstrated the safety of dofetilide. After pharmacological or electrical conversion of atrial fibrillation to sinus rhythm in these studies, the probability of remaining in sinus rhythm during the following year was 75%. Dofetilide has a single significant side effect: risk of developing torsade de pointes ventricular tachycardia. Therefore, dosage must be carefully adjusted to the length of QTc interval, calculated creatinine clearance and the presence of heart failure or recent infarction. In addition, treatment must be initiated in hospital with three days of continuous telemetry. Dofetilide can be co administered with digoxin and beta-blockers. Other anti-arrhythmic drugs, as well as drugs that interfere with the renal elimination or the metabolism of dofetilide, must be avoided. Dofetilide is an option when persistent atrial fibrillation is a clinical problem. In the setting of severe heart failure and large myocardial infarctions, only amiodarone and dofetilide have proven safety and dofetilide is a strong candidate for first choice treatment when the aim is to achieve sinus rhythm. PMID- 11060832 TI - Azimilide dihydrochloride: a new class III anti-arrhythmic agent. AB - Azimilide dihydrochloride (Stedicor) is a new class III anti-arrhythmic agent that is being developed by Proctor & Gamble to treat supraventricular and ventricular arrhythmias. Development of this agent is being undertaken due to the high prevalence of atrial fibrillation and the lack of satisfactory therapy for this arrhythmia, along with the desire to develop therapy to reduce the risk of life-threatening ventricular arrhythmias in patients following myocardial infarction. The mechanism of action of azimilide is to block both the slowly conducting (I(Ks)) and rapidly conducting (I(Kr)) rectifier potassium currents in cardiac cells. This differs from other class III agents that block I(Kr) exclusively or in combination with sodium, calcium, or transient outward (I(to)) potassium current channels. Azimilide is distinguished by a relative lack of reverse use-dependence, excellent oral absorption, no need for dose titration, an option for out-patient initiation, no need for adjustment associated with renal or liver failure and a lack of interaction with warfarin or digoxin. It carries some risk of torsade de pointes and rarely, neutropoenia. Azimilide has shown dose-related efficacy in prolonging the time to recurrence of atrial fibrillation. A large trial examining the impact of azimilide on mortality in high-risk patients following myocardial infarction has completed enrolment and should yield data in the next couple of years and further studies are planned. Even if this trial fails to show a survival benefit, a neutral effect on mortality will make the agent attractive for atrial arrhythmias. PMID- 11060833 TI - Evaluation of intravenous parecoxib for the relief of acute post-surgical pain. AB - Parecoxib is a prodrug of valdecoxib, which is a potent and selective inhibitor of COX-2. Intravenous preparation of parecoxib is in Phase III clinical trials for the management of acute and severe post-surgical pain. It is the only COX-2 inhibitor that is available in a parenteral formulation. Clinical results compare parecoxib with ketorolac, a NSAID, which is the only non-narcotic analgesic available in parenteral formulation that can be administered for the relief of moderate to severe acute pain. Pharmacokinetic studies have shown that parecoxib is converted to valdecoxib within a short time following administration by im. or iv. injection. In clinical trials, parecoxib compares favourably with ketorolac and produces less gastric or duodenal ulcers, the predominant adverse effect, than ketorolac. Parecoxib, thus, fulfils some of the desirable characteristics of an ideal non-narcotic analgesic for severe post-surgical pain and has application in other acutely painful conditions. Parecoxib is expected to be filed for approval before the end of 2000 and is expected to be introduced in the market in 2001. It has favourable prospects for a fair share of the post-surgical pain relief market which is valued at approximately US$ 1 billion for the year 2000. PMID- 11060834 TI - A brief history of flow cytometry and sorting. PMID- 11060835 TI - Principles of flow cytometry: an overview. PMID- 11060836 TI - Laser scanning cytometry. PMID- 11060837 TI - Principles of confocal microscopy. PMID- 11060838 TI - Optical measurements in cytometry: light scattering, extinction, absorption, and fluorescence. PMID- 11060839 TI - Flow cytometric fluorescence lifetime measurements. PMID- 11060840 TI - Principles of data acquisition and display. PMID- 11060841 TI - Time as a flow cytometric parameter. PMID- 11060842 TI - Protein labeling with fluorescent probes. PMID- 11060843 TI - Preparation of cells from blood. AB - Peripheral blood is a bountiful source of numerous cell types that are easily analyzed by flow cytometry for a variety of properties. The monodisperse nature of blood makes preparation of these cells relatively easy if the coagulation cascade is inhibited. For some studies the presence of serum can be a confounding factor, although this is often overcome by merely pelleting the cells from the serum and washing the remaining cells several times. Blood, particularly from humans and primates, should be considered highly infectious whether or not from a healthy donor. Therefore universal precautions should be followed at all times. Techniques and route of collection may have a profound influence on the condition and nature of the blood being obtained. Performing venipuncture with an extremely small gauge needle may disrupt cells and prevent satisfactory analysis. Venous blood and arterial blood may yield differing data. Even blood collection at different times of day may yield diurnal variations in some assays. The demands of each cell type and of each assay dictate specific preparation, fixation, storage, and staining protocols. However, the overall goal for preparation of cells from blood, for all of the assays, remains the same--collect and prepare the cells in such a manner that they accurately represent the in vivo state. PMID- 11060844 TI - Cell preparation for the identification of leukocytes. PMID- 11060845 TI - Strategies for cell permeabilization and fixation in detecting surface and intracellular antigens. PMID- 11060846 TI - Stoichiometry of immunocytochemical staining reactions. AB - This chapter reviewed and presented my biases about the factors that affect our ability to make quantitative measurements of epitope (for this chapter, equal to a protein) with monoclonal antibodies in a flow cytometric system. The discussion has illustrated that the chemical structure of the permeabilized cell and the affinity and specificity of the antibody are the "two" factors that are important. Pursuit of this discipline would be significantly enhanced with highly specific antibodies with a high affinity--higher than we have so far observed with the antibodies against cell cycle regulatory proteins. When moving to multiparametric space, optical systems that mask off interbeam cross talk, primary labeled antibodies, and rigorous use of fluorescence compensation is essential for the highest quality work. PMID- 11060847 TI - Standardization and quantitation in flow cytometry. PMID- 11060848 TI - Methods to identify mitotic cells by flow cytometry. PMID- 11060849 TI - Cell cycle kinetics estimated by analysis of bromodeoxyuridine incorporation. PMID- 11060850 TI - Flow cytometric analysis of cell division history using dilution of carboxyfluorescein diacetate succinimidyl ester, a stably integrated fluorescent probe. PMID- 11060851 TI - Antibodies against the Ki-67 protein: assessment of the growth fraction and tools for cell cycle analysis. PMID- 11060852 TI - Detection of proliferating cell nuclear antigen. PMID- 11060853 TI - Lymphocyte activation associated antigens. PMID- 11060854 TI - Analysis of mitochondria during cell death. PMID- 11060855 TI - Cytometry of caspases. PMID- 11060856 TI - Analysis of apoptosis in plant cells. PMID- 11060857 TI - Difficulties and pitfalls in analysis of apoptosis. PMID- 11060858 TI - Analysis of cell migration. PMID- 11060859 TI - Three-dimensional extracellular matrix substrates for cell culture. AB - In summary, the understanding of cell biology will be furthered as cell culture expands from 2-D to 3-D systems. In choosing which substrate, synthetic or biologically derived, is most well suited for a specific application, substrate composition and structure as well as cell type(s) must be carefully considered. In addition, optimization of seeding densities, medium conditions, growth factor supplements, and other culture parameters may be necessary. Finally, cytometric analyses of such 3-D culture systems will require concurrent innovations in 3-D imaging and methods for quantitating cell morphology, phenotype, and function. PMID- 11060860 TI - Three-dimensional imaging of extracellular matrix and extracellular matrix-cell interactions. AB - In summary, noninvasive and nondestructive imaging modalities such as reflection and autofluorescence can readily be used in conjunction with the 3-D optical sectioning capabilities of confocal and multiphoton microscopy to investigate biological processes within living systems. The elimination of specimen fixation and extensive processing reduces the possibility of structural artifacts and facilitates repeat observations within a single sample. Therefore, information representing up to four dimensions (x, y, z, and time) can be readily collected and reconstructed for purposes of visualization and/or quantitative analysis. An advantage of using the techniques described in this chapter is the possibility of performing quantitative measurement of cell size, surface area, volume, depth (in matrix), orientation, receptor density, as well as fluorescence-based indicators of phenotype and function. At present, we are effectively utilizing these techniques to study collagen fibrillogenesis and ECM assembly, structural aspects of ECM-based biomaterials, as well as cell interactions within 3-D matrices (e.g., migration). New insights provided by these techniques regarding ECM and ECM-cell signaling will further the understanding of tissue structure and function and contribute to the development of new and improved strategies for tissue repair, replacement, and maintenance. PMID- 11060861 TI - Cytometric analysis of cell contact and adhesion. PMID- 11060862 TI - Invadopodia: unique methods for measurement of extracellular matrix degradation in vitro. PMID- 11060863 TI - From the Einthoven galvanometer to the implantable loop recorder: revelations in store. PMID- 11060864 TI - The implantable loop recorder: the herald of a new age of implantable monitors. PMID- 11060865 TI - Assessment of the chronotropic response at the anaerobic threshold: an objective measure of chronotropic function. AB - The evaluation of the heart rate response to exercise is important for the diagnosis of chronotropic incompetence and the assessment of a rate responsive algorithm of sensor-controlled pacemakers. The aim of the present study was to examine a classification of the chronotropic response at an individually moderate exercise level. Sixteen pacemaker patients (patient group, age 62.9 +/- 7.6 years) with sick sinus syndrome and 15 age-matched healthy subjects (control group, age 57.6 +/- 9.4 years) underwent a maximum cardiopulmonary exercise test on a treadmill after a protocol with individually selected incremental steps. To analyze the patients' intrinsic heart rate response, the rate responsive mode of the pacemaker was switched off. Chronotropic incompetence was diagnosed in eight patients whose maximal heart rate was < 80% of the age-predicted heart rate. The heart rate at the anaerobic threshold was significantly lower in the chronotropically incompetent subgroup than in the chronotropically competent patients and the healthy subjects (85.9 +/- 6.6 beats/min vs 100.3 +/- 9.9 beats/min and 112.9 +/- 11.7 beats/min, respectively). The chronotropic slope of the heart rate reserve as a function of the metabolic reserve was significantly higher in the control group than in the patient groups with either mild or severe chronotropic incompetence (0.94 +/- 0.17 vs 0.64 +/- 0.08 and 0.43 +/- 0.14, respectively). Furthermore, the chronotropically incompetent response could be divided into a linear type with and without a threshold, an exponential, and a logarithmic type. The anaerobic threshold was an objectively detectable breakpoint at an individually moderate exercise level that could be used for characterization of chronotropic function. At the anaerobic threshold, a physiological heart rate response was about 220--age--50 beats/min. A deviation of more than 10 beats/min below this physiological value characterized chronotropic incompetence. PMID- 11060866 TI - Effects of a thin-sized lead body of a transvenous single coil defibrillation lead on ICD implantation. Kainox RV Study Group. AB - In the interest of patients receiving implantable cardioverter defibrillators (ICDs), the clinical benefits of newer and thinner transvenous defibrillation leads have to be determined. The aims of this study were to evaluate the ICD procedure duration and the frequency of lead dislocation at the 3-month follow-up of a new defibrillation lead with a thin-sized lead body and its conventional sized predecessor. The thin-sized single coil defibrillation lead (Kainox RV, Biotronik; lead body 6.7 Fr) was implanted in 61 patients and the conventional sized defibrillation lead (SPS, Biotronik; lead body 7.8 Fr) in 60 patients. Both leads were connected to a left-sided, prepectorally implanted Phylax ICD (Biotronik) with active housing. The lead implantation time and total procedure duration were determined. Lead implantation time was defined as the time from lead insertion to the end of the pacing measurements. The total procedure duration spanned skin incision to closure. The incidence of lead repositioning during the lead implantation time and during ventricular fibrillation conversion testing was also assessed. The frequency of lead dislocations was recorded at the 3-month follow-up. Mean lead implantation time and total procedure duration of the thin-sized lead (23 +/- 22 minutes 76 +/- 37 minutes) were not statistically different from the time needed for the conventional-sized lead (22 +/- 20 minutes 81 +/- 34 minutes). The number of lead repositionings during the lead implantation time was similar (thin-sized lead: 1.4 +/- 2.4; conventional-sized lead: 1.1 +/- 1.9). An additional lead repositioning was not necessary during ventricular fibrillation conversion testing in 93.4% of the patients with thin sized and in 94.4% with conventional-sized leads (not significant). At the 3 month follow-up, there were four (6.6%) lead dislocations in the thin-sized and four (6.7%) in the conventional-sized lead group. In conclusion, the down-sized lead body of the new defibrillation lead influenced neither ICD procedure duration nor the incidence of lead dislocation during follow-up. PMID- 11060867 TI - Influence of autothreshold sensing and sinus rate on mode switching algorithm behavior. AB - Mode switching is beneficial to pacemaker patients with paroxysmal atrial tachyarrhythmias. However, the optimal mode switching algorithm is still in evolution. Mode switching algorithms and atrial sensing circuitry can influence mode switching behavior. This study compared the mode switching behavior of four Medtronic, Inc. implantable devices: Thera DR model 7960 pacemaker, Kappa 700 model KDR701 pacemaker, Gem DR model 7271 dual chamber pacing defibrillator, and Jewel AF model 7250 dual chamber pacemaker atrial and ventricular defibrillator. The Thera and Gem DR use the same mean atrial rate mode switch algorithm. The Kappa and Jewel AF use four of seven short atrial intervals and an atrial fibrillation evidence counter algorithm, respectively. The Thera and Kappa devices use fixed gain sensing and the Gem DR and Jewel AF use autothreshold atrial sensing. Digitally recorded atrial electrograms from 52 episodes of human atrial fibrillation were fed into each device with differing simulated sinus rates before and after the atrial fibrillation. The percent of appropriate mode switching was highest for the Kappa 700 (94%) and lowest for the Thera (85%) (P = 0.046). The time to mode switching was significantly longer for the Thera and Gem DR compared to the Kappa 700 or Jewel AF (all P < 0.05). The time to mode switching was shorter for the Gem DR (9.0 +/- 1.6 s) using autothreshold atrial sensing than for the fixed gain Thera (11.1 +/- 2.1 s, P < 0.05). The mean atrial electrogram amplitude and cycle length were not correlated with the time to mode switching for any device. Faster sinus rates shortened the time to mode switching and prolonged the time to resynchronization in the two devices using the mean atrial interval algorithm. In conclusion, (1) mode switching function among these devices is influenced by algorithms and sensing circuitry, (2) the time to mode switching among these devices is influenced by the algorithm and use of autothreshold atrial sensing, and (3) the sinus rate before and after episodes of atrial fibrillation greatly influences the times to mode switching and resynchronization in devices using the mean atrial interval algorithm. PMID- 11060868 TI - Electroanatomic mapping to identify breakthrough sites in recurrent typical human flutter. AB - The accuracy of conventional techniques in localizing previous radiofrequency (RF) ablation sites and thus breakthrough sites of recurrent atrial flutter is somewhat limited. We investigated the role of electroanatomic mapping for identifying breakthrough sites or "gaps" at the tricuspid annulus and inferior vena cava (IVC)/eustachian ridge isthmus to help RF ablation in patients with recurrent typical flutter. Twelve patients (8 men, 4 women, age 63 +/- 10 years) with recurrent typical atrial flutter were included in the study. An electroanatomic mapping system (CARTO) was used to create a voltage map and activation and propagation patterns in the right atrium. Detailed voltage, activation, and propagation mapping of the tricuspid annulus and IVC/eustachian ridge isthmus allowed precise identification of gaps in all 12 patients at the tricuspid annulus (eight sites), IVC ridges (two sites), mid-isthmus region (one site), and tricuspid annulus and IVC ridges (one site). Radiofrequency energy directed at these sites eliminated atrial flutter in all 12 patients, confirmed by noninducibility of atrial flutter and demonstration of conduction block during atrial pacing on either side of the lesion lines. During a mean follow-up of 14.8 +/- 3.5 months (range 8-19 months), paroxysmal atrial flutter recurred in only one patient and was subsequently treated with amiodarone, although this had been ineffective prior to ablation. Electroanatomic mapping can precisely identify gaps in the lesion line responsible for breakthrough of recurrent typical atrial flutter at the tricuspid annulus and at the IVC/eustachian ridge isthmus. These sites can be targeted with RF ablation with a high degree of success. PMID- 11060869 TI - Differentiating the ligament of Marshall from the pulmonary vein musculature potentials in patients with paroxysmal atrial fibrillation: electrophysiological characteristics and results of radiofrequency ablation. AB - It was reported that paroxysmal atrial fibrillation (PAF) can be initiated by ectopic atrial beats originating from the pulmonary vein (PV) or left atrial tract (LAT) within the ligament of Marshall (LOM). The aim of this study was to differentiate the LAT from the PV potentials, and to investigate the results of radiofrequency ablation guided by these potentials. Ten patients (age 60 +/- 12 years) with PAF who had a recording of double potentials (DPs) in or around the left PV were included. Group I had five patients with the second deflection of DPs (D2) due to activation of the LAT, and Group II had five patients with D2 due to activation of the PV musculature. There were no significant difference in the isoelectric interval between DPs, the activation time, and amplitude of D2 between Groups I and II. During distal coronary sinus (CS) pacing, the CS ostium (CSO) to D2 interval was shorter compared with that during sinus rhythm in Group I (39 +/- 19 vs 71 +/- 25 ms, P = 0.04), but was longer in Group II (96 +/- 16 vs 44 +/- 19 ms, P = 0.04). During ectopic activation, three patients in Group I, but no Group II patients, had transformation of recorded DPs into triple potentials. Radiofrequency ablation guided by the earliest activation of the LAT potential was performed with transient suppression of PAF, but ablation guided by the earliest activation of the PV potentials had a high success rate in eliminating PAF. In conclusion, differentiating the LAT from the PV potentials for initiation of PAF is feasible by an electrophysiological approach, and may be important for radiofrequency ablation of PAF. PMID- 11060870 TI - Closed-loop stimulation using intracardiac impedance as a sensor principle: correlation of right ventricular dP/dtmax and intracardiac impedance during dobutamine stress test. AB - Changes of the unipolar right ventricular impedance during the cardiac cycle are related to the changing content of blood (low impedance) and tissue (high impedance) around the tip of the pacing electrode. During myocardial contraction, the impedance continuously increases reaching its maximum in late systole. This impedance increase is thought to correlate with right ventricular contractility, and thus, with the inotropic state of the heart. In the new Inos2 DDDR pacemaker, integrated information from the changing ventricular impedance (VIMP) is used for closed-loop regulation of the rate response. The aim of this study was to analyze the effect of increasing dobutamine challenge on RV contractility and the measured impedance signals. In 12 patients (10 men, 68 +/- 12 years) undergoing implantation of an Inos2 DDDR pacemaker (Biotronik), a right ventricular pigtail catheter was inserted for continuous measurements of RV-dP/dtmax and simultaneous VIMP signals during intrinsic and ventricular paced rhythm. Then, a stress test with a stepwise increase of intravenous dobutamine (5-20 micrograms/kg per min) was performed. To assess the relationship between RV contractility and measured sensor signals, normalized values of dP/dtmax and VIMP were compared by linear regression. There was a strong and highly significant correlation between dP/dtmax and VIMP for ventricular paced (r2 = 0.93) and intrinsic rhythm (r2 = 0.92), although the morphologies of the original impedance curves differed quite substantially between paced and intrinsic rhythm in the same patient. Furthermore, VIMP correlated well with sinus rate (r2 = 0.82), although there were at least four patients with documented chronotropic incompetence. We conclude, that for intrinsic and ventricular paced rhythms sensor signals derived from right ventricular unipolar impedance curves closely correlate with dP/dtmax, and thus, with a surrogate of right ventricular contractility during dobutamine stress testing. Our results suggest that "inotropy-sensing" via measurement of intracardiac impedance is highly accurate and seems to be a promising sensor principle for physiological rate adaptation in a closed-loop pacing system. PMID- 11060871 TI - Clinical evaluation of a pacemaker algorithm that adjusts the pacing rate during sleep using activity variance. AB - Even though rate responsive pacemakers are able to regulate pacing rates based on sensor activity, they are set with a minimum rate that is not adjusted to provide rate decreases during sleep. The aim of this study was to evaluate the performance of the "Sleep Rate" feature, as compared to patient diaries and a validated method that identifies sleep from wrist actigraphy. In 19 patients (15 men; age 69 +/- 8 years) with Pacesetter Trilogy DR+ pacemakers, the base rate and the sleep rate were set to 80 and 50 ppm, respectively. When the patients returned 2 days later, data recorded by the pacemaker and wrist actigraph were analyzed to find the agreement in corresponding sleep/wake periods. In 17 (89%) patients, the pacemaker went into the sleep mode. The total sleep time derived from actigraphy significantly exceeded the time during which the pacemaker was in sleep mode (1156.8 +/- 83.4 vs 307.3 +/- 77.2 minutes). Frequent reversions out of the sleep mode limited the total sleep time derived from the pacemaker. Cumulative analysis of the pacemaker data showed that the maximum time in the sleep mode was 78 minutes, and exceeded 1 hour in six instances, 30 minutes in 32 instances, and 15 minutes in 83 instances. Epoch by epoch comparisons revealed a good agreement (93.6 +/- 1.8%) during wakefulness between the corresponding actigraph and pacemaker epochs. However, only 24.6 +/- 3.7% of the corresponding epochs during sleep were identical, and the overall agreement was 54.4 +/- 3.7%. Except for one patient who reported palpitations, patients did not suffer from a pacemaker rate change. The Sleep Rate feature provides rate reduction during sleep, while assuring rapid frequency response during physical activity. However, the current algorithm does not allow long periods of slow pacing rate during continuous sleep, possibly due to its conservative design and the presence of movement arousals, which has to be improved in future generation algorithms. PMID- 11060872 TI - Electromagnetic interference of an implantable loop recorder by commonly encountered electronic devices. AB - Electromagnetic interference of pacemaker systems has been well established and can lead to an inappropriate function of these devices. Recently, an implantable loop recorder (ILR) (REVEAL, Medtronic Inc.) has been introduced to evaluate the possible arrhythmic etiology of patients with recurrent syncope. We evaluated the interference of this device in two patients with implantable ILR and in three nonimplanted ILRs with four electromagnetic sources: cellular phones (GSMs), electronic article surveillance systems (EASs), metal detector gates (MDGs), and magnetic resonance imaging (MRI). The GSM did not affect appropriate function of the ILR whereas radiofrequency (RF) EAS could interfere with normal function in implanted and nonimplanted systems. The MDG had no influence on ILR function. The magnetic field induced by the MRI resulted in an irreversible error in one nonimplanted ILR. Therefore, although interference between electromagnetic sources and ILRs appears to be rare in our study, physicians should be aware of possible malfunctioning of these devices. PMID- 11060873 TI - The Bayesian approach improves the electrocardiographic diagnosis of broad complex tachycardia. AB - Despite numerous attempts at devising algorithms for diagnosing broad complex tachycardia (BCT) on the basis of the electrocardiogram (ECG), misdiagnosis is still common. The reason for this may lie with difficulty in implementing existent algorithms in practice, due to imperfect ascertainment of ECG features within them. An attempt was made to approach the problem afresh with the Bayesian inference by the construction of a diagnostic algorithm centered around the likelihood ratio (LR). Previously studied ECG features most effective in discriminating ventricular tachycardia (VT) from supraventricular tachycardia with aberrant conduction (SVTAC), according to their LR values, were selected for inclusion into a Bayesian diagnostic algorithm. A test set of 244 BCT ECGs was assembled and shown to three independent observers who were blinded to the diagnoses made at electrophysiological study. Their diagnostic accuracy by the Bayesian algorithm was compared against that by clinical judgement with the diagnoses from EPS as the criterial standard. Clinical judgement correctly diagnosed 35% of SVTAC, 85% of VT, and 47% of fascicular tachycardia. In comparison, by the Bayesian algorithm devised, 52% of SVTAC, 95% of VT, and 97% of fascicular tachycardia were correctly diagnosed. The Bayesian algorithm devised has proved to be superior to the clinical judgement of the observers who participated in this study, and theoretically will obviate the problem of imperfect ascertainment of ECG features. Hence, it holds the promise for being an effective tool for routine use in clinical practice. PMID- 11060874 TI - Paradoxical AV delay shortening of a pacemaker. AB - Paradoxical shortening of the paced AV delay (atrial paced-ventricular paced or Ap-Vp interval) was observed at rest in the DDD and DDDR modes in three patients with implanted CPI Vigor DR pacemakers programmed with a long AV delay and a relatively narrow difference between the lower and upper rates. This behavior is related to the VA extension algorithm designed to prevent the sensor-driven atrial pacing rate from exceeding the programmed upper rate whenever a sensed conducted QRS complex continually follows an atrial stimulus. We found that this algorithm also becomes manifest at rest and may cause shortening of the Ap-Vp intervals. The VA extension algorithm is best conceptualized in terms of a separate atrial upper rate that functions on exercise and at rest. The atrial and ventricular upper rates are equal but the atrial upper rate is initiated by an atrial-paced or sensed event and the ventricular upper rate is initiated by a paced or sensed ventricular event. Under certain circumstances delay in the release of the atrial stimulus Ap to conform to the atrial upper rate interval produces variable abbreviation of the paced AV (Ap-Vp) delays with resultant variation in the duration of the atrial escape intervals despite fundamental ventricular-based lower rate timing. PMID- 11060875 TI - Heart rhythm during syncope and presyncope: results of implantable loop recorders. AB - Ambulatory ECG monitoring in patients with recurrent syncope is nondiagnostic in the majority of cases. Recently, an ECG implantable loop recorder (ILR) has been introduced. The ILR performs continuous ECG monitoring over a period of at least 14 months. From February 1997 to September 1999, 35 patients underwent implantation of an ILR. During a mean follow-up of 11 +/- 8 months, 24 (69%) patients had recurrent syncope or presyncope events. Four (11%) patients were not capable of activating the ILR to save the event. A symptom-rhythm correlation could be studied in 20 (83%) of 24 patients. Forty of 44 recurrences were captured by the ILR. There were 14 (40%) patients with at least one syncopal episode. An arrhythmic cause for syncope was found in eight of them (bradycardia in four and tachycardia in four). In the other six patients the heart rhythm was normal. In 17 (49%) patients with 1-year follow-up, the mean syncope event rate 12 months before ILR implantation was 4.7 +/- 2.4, whereas the mean syncope event rate 12 months after ILR implantation was 1.3 +/- 0.7 (P < 0.01). Resolution of symptoms was observed in 6 (17%) patients. These patients were significantly younger than patients without resolution (50 +/- 18 vs 69 +/- 14 years, p < 0.01) and five were women. Three (9%) patients died during follow-up, all of them were noncompliant during their follow-up. In conclusion, the ILR made symptom-rhythm correlation possible in 83% of patients with recurrent syncope. Syncope recurrences decreased significantly after implantation of the device, especially in the younger patients. Noncompliant patients had a high mortality rate. PMID- 11060876 TI - Time dependent changes in duration of ventricular repolarization after AV node ablation: insights into the possible mechanism of postprocedural sudden death. AB - Although effective, there is a disturbing incidence of sudden death after AV node ablation. The mechanism may be related to proarrhythmia associated with prolongation in ventricular repolarization from the sudden decrease in heart rate. To examine this issue, we studied 15 patients undergoing complete radiofrequency ablation of the AV node for rapid atrial arrhythmias. Twelve-lead ECGs of paced rhythms at rates of 60, 80, 100, and 120 beats/min were recorded at time points of 30 minutes, 24 hours, 1 week, and 1 month after ablation. The QT interval was measured in the limb and precordial leads with the best T wave offset. The change in the QT interval (delta QT) relative to the measurement at 30-minute postablation was calculated. For comparison, a similar procedure was performed on patients receiving pacemakers for primary bradycardia (n = 5). The mean QT interval at 60 beats/min, 30-minutes postablation was significantly longer than at time points thereafter (482 +/- 39 vs 446 +/- 28 ms at 1 month, limb leads, for example, P < 0.05). Analysis of delta QT revealed a significant shortening of the QT interval at nearly every paced rate at every time point relative to the value at 30-minute postablation. The QT intervals shortened and stabilized after 24 hours. Neither the QT interval nor delta QT changed significantly in patients paced for primary bradycardia. We conclude that there is a relative increase in the duration of ventricular repolarization after AV node ablation, which then decreases and stabilizes after 24 hours. Such changes are not seen in patients being paced for primary bradycardia. This data is consistent with the hypothesis that sudden death after AV node ablation may be related to proarrhythmia from prolonged ventricular repolarization. PMID- 11060877 TI - How can we identify the best implantation site for an ECG event recorder? AB - The aim of this study was to show how to find the preferable implantation site for an ECG event recorder (ECG-ER). We compared the quality of bipolar ECG recordings (4-cm electrode distance, vertical position) in 65 patients at the following sites: left and right subclavicular, left and right anterior axillary line (4th-5th interspace), left and right of the sternum (4th-5th interspace), heart apex, and subxyphoidal. The results were compared to the standard ECG lead II. In 30 patients, an additional comparison between vertical and horizontal ECG registrations was done using the same sites. ECG signals in five patients were compared positioning the electrodes towards the skin with turning them towards the muscle during ECG-ER implantation. The best ECG quality (defined as highest QRS amplitude, best visible P wave and/or pacemaker spike, best measurable QRS duration, and QT interval) and best agreement with the standard lead II was found in 68% on the left of the sternum, significantly less often (P < 0.001) on the right of the sternum (14.1%), left subclavicular (6.9%), apical (5.5%) and subxyphoidal (4.2%). A significantly higher QRS amplitude was measured and the P wave was more often visible in the vertical electrode position than in the horizontal position. In all five ECG-ER patients, there was a good agreement between the bipolar surface ECG at the implantation site and ECG-ER stored signals. A significant noise signal occurred in all five patients when the ECG-ER was implanted with electrodes towards the muscle. A P wave was visible in only three of those patients, but there was an insignificantly higher QRS amplitude than in ECG-ERs implanted with electrodes towards the skin. From these results, it can be concluded that the best implantation site for an ECG-ER is right or left of the sternum, positioning the electrodes vertically and towards the skin. PMID- 11060878 TI - Women and arrhythmias. PMID- 11060879 TI - Double potentials in the right superior pulmonary vein. PMID- 11060880 TI - Paroxysmal supraventricular tachycardia in identical twins with the same left lateral accessory pathways and innocent dual atrioventricular pathways. AB - We report on 16-year-old, female identical twins who both have atrioventricular reentrant tachycardia caused by the same left lateral atrioventricular accessory pathway. The Kent pathway in twin A was a unidirectional retrograde accessory pathway. A manifest Kent pathway was demonstrated in twin B. Both pathways were successfully ablated by radiofrequency (RF) energy and without recurrence. In addition, innocent dual AV nodal pathways were shown in both patients. These findings suggest that genetic factors may play a role in the pathogenesis of the formation of accessory atrioventricular pathways and dual AV nodal pathways. PMID- 11060881 TI - Concealed accessory pathway manifesting clinically only after pacemaker implantation. AB - A 66-year-old man with coronary artery disease and persistent left superior vena cava received a DDDR pacemaker for symptomatic 2:1 heart block. There was no previous history of tachyarrhythmias. Endless loop tachycardia and repetitive nonreentrant ventriculoatrial synchrony occurred afterwards and were triggered by a late coupled atrial premature beat. ECGs suggested a concealed left posterior accessory pathway that was confirmed during electrophysiological study. Effective palliation was achieved with extension of the PVARP and enabling noncompetitive atrial pacing operation. PMID- 11060882 TI - Device eccentricity: postmagnet behavior of DDDR pacemakers with automatic threshold tracking. AB - The normal pacing system function associated with the AutoCapture Pacing System algorithm triggered concern on the part of the clinical staff caring for the patient when the initiation of a threshold search sequence was detected during a transtelephonic follow-up evaluation. The analysis of the rhythm demonstrates that the behavior of the system is normal and consistent with the design of the AutoCapture algorithm. As a variety of new algorithms are introduced, similar unexpected but normal behaviors can be anticipated. PMID- 11060883 TI - Insertable loop recorder in unmasking the cause of syncope. AB - During the last few years closed loop recorders are being used increasingly in the investigation of unexplained syncope. Our patient had a 5-year history of pre and syncopal episodes in which the usual invasive and noninvasive tests were nondiagnostic. Finally, a loop recorder was implanted that revealed the cause of the syncope a few days after implantation: a fast, sustained, ventricular tachycardia originating from the right ventricular outflow tract. Loop recorders will undoubtedly contribute toward the decrease in the percentage of undiagnosed syncope cases and possibly to the reduction of the investigation cost. PMID- 11060884 TI - Demonstration of myocardial ischemia by an internal loop recorder. AB - An internal loop recorder (ILR) implanted to evaluate syncope was activated during an episode of chest pain. Analysis of the recorded event revealed a marked increase in the amount of ST-segment depression over baseline. In addition to rhythm analysis, the ILR may be able to assess myocardial ischemia. Further refinements of filtering may make analysis more accurate. PMID- 11060885 TI - [The intraspecific differentiation of the burrow tick Ixodes crenulatus (Ixodidae)]. AB - In four extensive disjunct areas of the distribution range of Ixodes crenulatus Koch, 1844 complexes of samples in 8 locations, and separate samples in two locations have been studied (fig.). Morphological characters (sizes of organs of idiosome, gnathostoma, legs, as well as some proportions of organs), which show statistically significant differences between complexes of samples on all corresponding stages of ontogenesis were revealed (tabl. 1, 2). Statistically significant differences were determined by Student's criterion (table. 3, 4). We use the term "transit" characters to denote these characters if they have to addition a similar tendency at all stages of ontogenesis. Complexes of samples showing statistically significant differences of transit characters are considered by us as morphotypes. Apparently these differences of morphotypes were formed evolutionally. In European disjunct area morphotypes divided into two groups on the basis of the degree of differences: western (A, B), and eastern (C, D, E). Degree of differences of morphotypes within each group is low, whereas one between of these two groups is high and corresponds to differences of morphotypes A, B from all Asian morphotypes (F, G, H, I, J), and of morphotypes C, D, E from Asian ones in mountain disjunct areas I. crenulatus (G, H, I, J). At the same time Eastern-European complex of morphotypes (C, D, E) is morphologically similar to the morphotype from north Kazakhstan disjunct area (F). All Asian morphotypes have high degree of differences one from the other. Species of the genus Marmota are initial hosts of I. crenulatus, they retain the main role as hosts in Eastern European and Asian morphotypes: C, D, E, F--M. bobac bobac (Muller, 1776), G--M. baibacina centralis (Thomas, 1909), I, J--M. sibirica sibirica (Radde, 1862). The most aberrant in all characters morphotypes are the ones inhabiting European areas (A, B), where marmots are exterminated. These morphotypes parasitize on hibernating carnivores. PMID- 11060886 TI - [The longevity of Leptopsylla segnis fleas (Siphonaptera: Leptopsyllidae)]. AB - In experiments, the mean life duration of fleas Leptopsylla segnis on white mice (abundance of fleas within natural limits, up to 10 fleas per mouse) was 22.7 days in females and 18.8 day in males. Maximum life duration was 51 and 37 days respectively. In cases, when the initial numbers of fleas were 20 and 28-34 fleas, the duration of life was decreased. The maximum limit decreased greater than the mean duration of life. A survival dynamics of fleas depended upon the flea number. It was found out, that in cases of high abundance of fleas in the beginning of experiments, the mortality rate of males was lower than in females. During the stay on a host the fleas lost gradually an ability to endure a starvation. Possible mechanisms of the regulation of flea abundance are discussed. PMID- 11060887 TI - [A karyotypic analysis of 2 species of blackflies in the genus Prosimulium (Diptera: Simuliidae) from the Kola Peninsula and Kamchatka]. AB - Significant karyotypic differences between the triploid (3n = 9) Prosimulium macropygum Lundstrom and the diploid (2n = 6) P. macropygum ventosum Rubzov have been found. It is suggested to consider the latter taxon as an independent species P. ventosum. PMID- 11060888 TI - [The current composition of the subfamily Encotyllabinae (Monogenoidea: Capsalidae)]. AB - The historical account and recent taxonomic consideration of the monogenean subfamily Encotyllabinae are given. Main criteria used for diagnostics of the subfamily and its genera are discussed. PMID- 11060889 TI - [The cestode fauna of the fam. Hymenolepididae in the anatine birds of Chukotka. The genus Dicranotaenia]. AB - There are 5 species of the genus Dicranotaenia recorded in the North-west of Chukotka: Dicranotaenia coronula (?) (Dujardin, 1845), D. fallax (Krabbe, 1869), D. parvisaccata (Shepard, 1943), D. sacciperum (Mayhew, 1925) and D. clangulae sp. n. Brief characteristic of the new species. Rostellar hooks 24-28, their total length 17-20 mkm, blade 10-12 mkm, guard processus 7-9 mkm, basis 13-15 mkm. Blade is parallel to basis, significantly longer than guard processus, hook form of aploparaksoid type. Cirrus pouch with s-like bend and distal enlargement (as in D. fallax). Cirrus almost cylindrical, 115-136 mkm in length, diameter at base 20-23 mkm, distal diameter 13-16 mkm. Internal acessory sac (SAI) situated anterior and dorsal from base of cirrus, gradually attenuate to distal end, length of SAI 27-30, diameter at base 30-32. Descriptions of adult forms and metacestodes of four other species are given. PMID- 11060890 TI - [The morphological variability of the maritae of Phyllodistomum umblae and Phyllodistomum folium (Trematoda: Gorgoderidae) from fish in the Lake Baikal basin]. AB - The host variability of adults of Phyllodistomum umblae and P. folium in fishes of the Baikal region has been studied. The absence of considerable geographical variability of P. umblae in Coregonus lavaretus (Lake Baikal, Lake Storsjon in Sweden) has been shown. The complexes of most stable (the size of acetabulum and eggs, location of acetabulum, ovary and vitelline bodies) and variable (the distance from oral sucker to intestinal bifurcation, distance from intestinal back edge to body end, width of seminal vesicle) features of trematodes of the genus Phyllodistomum have been determined. PMID- 11060891 TI - [The distribution of entomopathogenic nematodes of the families Steinernematidae and Heterorabditidae in Russia and their morphological characteristics]. AB - The data on a distribution of nematode species of the families Steinernematidae and Heterorabditidae in the northern and southern regions of the Russian Federation are given. Morphological characters of geographic isolates of some nematode species are not different, while their invasive activities in dependence of a temperature are different. Steinernema feltiae occurs in all climate zones, S. carpocapsae is restricted in distribution, nematode species of the family Heterorabditidae were found in southern regions only. PMID- 11060892 TI - [The morphology of the macrogametes and oocysts of Alveocystis intestinalis (Sporozoa: Coccidia)]. AB - Macrogametes and oocysts of the coccidium Alveocystis intestinalis parasitizing in priapulids have been examined. It is shown, that the macrogametes of this species have small granules of glycoproteinaceous nature. These granules are wall forming bodies. It was considered these bodies are absent in A. intestinalis. The oocysts in the investigated material contained mainly 4 sporozoites. The oocysts with 8 sporozoites, which were observed to be most frequent in the materials used for the original description of A. intestinalis, is not a standard for this species. Therefore the joining of A. intestinalis and Pfeifferinella gugleri into one genus based on a similarity of their oocyst structure is incorrect. This conclusion is also supported by the long evolutionary and ecological distances between hosts of these species. PMID- 11060893 TI - [The independence of the family Channaculinidae (Crustacea: Copepoda: Cyclopoida)]. AB - New family name Pillainidae Kazatchenko fam. nov. is suggested instead of family name Channaculinidae. The keys of the families and genera of fish parasitic copepods of suborder Cyclopoida are given. PMID- 11060894 TI - [Planorbinae snails--obligate intermediate hosts of paramphistomatid trematodes in the central Polesye of Ukraine]. AB - The data on species composition, ecology and distribution of snails of the subfamily Planorbinae (tribes Planorbini, Segmentinini) in water basis of Tsentral'noye Polesye are given. In 13 species belonging to the genera Planorbis, Anisius, Choanomphalus, Segmentina, Hippeutis, the parthenitae and larvae of Paramphistomum ichikawai Jamaguti has been recorded. Infection rate varies from 0.5 to 18.5%. PMID- 11060895 TI - [The biology of the trematodes Nenimandijea kashmirensis and Pleurogenoides medians (Pleurogenidae)--parasites of frogs in the Maritime Territory]. AB - The experimental study of life cycles of the trematodes Nenimandijea kashmirensis Kaw, 1950 and Pleurogenoides medians Olsson, 1876 was carried out. It was found out, that their life cycles include: the first intermediate host--the mollusc Boreoelona contortrix ussuriensis, the second intermediate host--dragonfly larvae of the genus Cordulia, and the final host--the frogs Rana nigrimaculata and R. semiplicata. Based on obtained data it is suggested, that Pleurogenoides japonicus (Yamaguti) should not be considered as a separate species. PMID- 11060896 TI - Mechanism and signal transduction of 14 (R), 15 (S)-epoxyeicosatrienoic acid (14,15-EET) binding in guinea pig monocytes. AB - 14(R), 15(S)-epoxyeicosatrienoic acid (14,15-EET) is a cytochrome P-450 monooxygenase (epoxygenase) metabolite of arachidonic acid (AA). In this study, we have identified a population of specific high affinity binding sites for 14,15 EET in the guinea pig mononuclear (GPM) cells. The results of competition studies showed that 14(R), 15(S)-EET was an effective competing ligand with a Ki of 226.3 nM followed by 11(R), 12(S)-EET, 14(S), 15(R)-EET, 14,15 thia(S)-ET, and 14,15 aza(N)-ET. The binding was sensitive to various protease treatments suggesting that the binding site is protein in nature. Cholera toxin (CT) and dibutyryl cAMP attenuated 14,15-EET binding in GPM cells. Mean binding site density (Bmax), decreased 32.0% and 19.1% by the pretreatment with cholera toxin (200 micrograms/ml) and dibutyryl cAMP (100 nM), respectively, without changing the dissociation constant. A specific protein kinase A (PKA) inhibitor, H-89, but not the PKC inhibitor K252a reversed the down regulation of 14,15-EET receptor binding caused by dibutyryl cAMP in GPM cells. Thus, the results sug-gest that the specific binding site of 14,15-EET in GPM cells be associated with a receptor that could be down regulated through an increase in intracellular cAMP and activation of a PKA signal trans-duction. We propose that the signal transduction mechanism begins with the binding of 14,15-EET to its receptor that leads to increase intracellular cAMP levels and the activation of PKA, and finally, with the down regulation of 14,15-EET receptor binding. PMID- 11060897 TI - Isoprostanes induce plasma extravasation in rat skin. AB - Isoprostane E2 (8-iso PGE) and isoprostane F2 alpha (8-iso PGF) contribute to numerous vascular, proinflammatory, and nociceptive functions. The underlying mechanisms for many of their actions are still under investigation. We examined the ability of isoprostanes to promote cutaneous inflammation using the Evan's blue dye method. Our data show that 4 micrograms subcutaneously (s.c.) injected 8 iso PGE or 8-iso PGF induced plasma extravasation in glabrous rat skin. Dye extravasation was also elicited in hairy skin after injections of 8-iso PGE, but not after 8-iso PGF. Isoprostane-evoked dye extravasation can be reduced by pretreatment with both the S+ and R- isomers of the cyclooxygenase (COX) inhibitor ibuprofen (30 mg/kg intraperitoneally), indicating perhaps a nonspecific inhibition; pretreatment with ketorolac (1 and 10 mg/kg i.v.) was without effect. Unlike isoprostane-induced cutaneous nociceptor sensitization, which is blocked in a stereospecific and dose-dependent manner by COX-inhibitors, the effect of these drugs on isoprostane-induced cutaneous plasma extravasation is less consistent. We conclude that at least a large component of the isoprostane effect on cutaneous plasma extravasation is COX-independent. PMID- 11060898 TI - Regulation of leptin release and lipolysis by PGE2 in rat adipose tissue. AB - The role of eicosanoids formed by adipose tissue from rats was examined in the presence of the specific cyclooxygenase-2 inhibitor NS-398. This agent totally blocked the release of prostaglandin E2 (PGE2) by rat adipose tissue over a 24-h incubation in primary culture. The final concentration of PGE2 after 24 h was 12 nM, and half-maximal inhibition of PGE2 formation required 35 nM NS-398. While inhibition of PGE2 formation by NS-398 had no effect on basal leptin release or lipolysis, it enhanced the lipolytic action of 10 nM isoproterenol by 36%. The in vivo administration of PGE2 doubled serum leptin. PGE2 also directly stimulated leptin release by rat adipose tissue incubated in the presence of 25 nM dexamethasone, which inhibited endogenous PGE2 formation by 94%. The inhibition of lipolysis as well as the stimulation of leptin release by PGE2 were mimicked by N6-cyclopentyladenosine (CPA). These data indicate that exogenous PGE2 can stimulate leptin release by adipose tissue when the basal formation of PGE2 is blocked by dexamethasone. However, while the endogenous formation of PGE2 does not appear to regulate basal lipolysis or leptin release, it may play a role in the activation of lipolysis by catecholamines. PMID- 11060899 TI - Oxidative response and membrane modification of diabetic platelets challenged with PAF. AB - Alterations in the functional activities of platelets (PLT) in type I diabetes have been widely observed. These changes play a key role in the development of cardiovascular complications in diabetes. Various functional activities of PLT are the result of the interaction of numerous stimuli with PLT plasma membrane. This study was designed to evaluate the oxidative response and membrane modifications of diabetic PLT stimulated by platelet activating factor (PAF). The oxidative response was assessed by employing luminol- and lucigenin-amplified chemiluminescence. Luminol-amplified chemiluminescence is sensitive to the release of hydrogen peroxide whereas lucigenin-amplified chemiluminescence is sensitive to the production of superoxide anion. Membrane fluidity and polarity were studied using fluorescence spectroscopy. Membrane fluidity was investigated by measuring steady-state fluorescence anisotropy of 1-[4-trimethylammonium phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH) and membrane polarity was studied by measuring the steady-state fluorescence emission and excitation spectra of 2 dimethylamino[6-lauroyl]-naphthalene (Laurdan). The diabetic group consisted of 20 type I diabetic children with good metabolic control. Our results show a significant decrease in the luminol- and lucigenin-amplified chemiluminescence of PAF stimulated PLT in the diabetic group with respect to controls. These data indicate a decrement in the release of reactive oxygen species by diabetic PLT. We observed a significant increase in steady-state fluorescence anisotropy of diabetic PLT membrane that reflects a decrease in membrane fluidity. Laurdan showed a blue shift of the fluorescence emission and excitation spectra in diabetic PLT with respect to the control group, indicating a decrease in membrane polarity. The addition of PAF to PLT induced a red shift of Laurdan spectra in both groups, indicating an increase in membrane polarity. Our study [table: see text] demonstrates an altered oxidative response to PAF stimulation of diabetic PLT, probably due to altered generation or handling of reactive oxygen species, and alterations in the physico-chemical properties of the plasma membrane which could influence various functional activities of PLT. PMID- 11060900 TI - Topical application of a selective cyclooxygenase inhibitor suppresses UVB mediated cutaneous inflammation. AB - Ultraviolet B (UVB) radiation causes much of the cutaneous damage after both acute and long-term exposure, and is also the most important etiologic agent in human skin cancer. UVB exposure initially induces an inflammatory response characterized by edema, dermal infiltration of leukocytes, sunburn cell formation, as well as the induction of cyclooxygenase-2 (COX-2) gene expression and subsequent increase in the production and release of prostaglandins. This process of inflammation induced by UVB exposure has been linked to tumor formation. Recently, a specific COX-2 inhibitor, Celecoxib, was developed, which inhibits COX-2-induced inflammation without inhibiting the cytoprotective function of cyclooxygenase-1 (COX-1). The present study compared the effects of topical treatment with Celecoxib (a specific COX-2 inhibitor) and Ibuprofen (a nonspecific COX inhibitor) on the acute UVB-induced cutaneous inflammatory response. We show that the specific inhibition of COX-2 effectively reduced many parameters of UVB-mediated inflammation, including edema, dermal neutrophil infiltration and activation, prostaglandin E2 (PGE2) levels and the formation of sunburn cells. By inhibiting this inflammatory response, topical Celecoxib treatment may ultimately be effective in preventing UVB-induced tumor development in the skin. PMID- 11060901 TI - Mediation by platelet-activating factor of 12-hydroxyeicosatetraenoic acid induced cytosolic free calcium concentration elevation in neutrophils. AB - 12(R)-hydroxyeicosatetraenoic acid (HETE) shows biphasic increase in cytosolic free calcium concentration ([Ca2+]i) in rabbit and human neutrophils; the initial transient phase and the continuous falling phase. 12(S)-HETE was less potent in both species. BN50739, a platelet-activating factor (PAF) receptor antagonist, inhibited both phases of 12(R)-HETE-induced [Ca2+]i rise but did not affect leukotriene B4 (LTB4)-induced [Ca2+]i rise. N-tosyl-L-phenylalanine chloromethyl ketone (TPCK), a PAF synthesis inhibitor, and manoalide, a phospholipase A2 inhibitor, reduced 12(R)-HETE-induced [Ca2+]i rise. These blockers inhibited the continuous phase of [Ca2+]i rise induced by N-formyl-methionyl-leucyl phenylalanine (FMLP) with little effect on the initial phase. It had no significant effect on LTB4-induced [Ca2+]i rise. SC-41930, a LTB4-receptor antagonist, did not block 12-HETE-induced [Ca2+]i rise. In 12(R)-HETE-, FMLP- and LTB4-stimulated cells, accumulations of cell-associated PAF and released PAF were detected but not in unstimulated cells. BN50739 did not affect the accumulation of cell-associated PAF and release of PAF in 12(R)-HETE-stimulated cells. These results suggest that 12(R)-HETE-induced and partially, FMLP-induced, but not LTB4 induced [Ca2+]i rise are mediated by PAF, which is produced and released by stimulation of the cells by 12(R)-HETE and FMLP, respectively. PMID- 11060902 TI - Difference in urinary LTE4 and 11-dehydro-TXB2 excretion in asthmatic patients. AB - Bronchoconstrictor cysteinyl leukotrienes (LT) and thromboxane (TX) A2 have been implicated in the pathogenesis of asthma. Determination of urinary leukotriene E4 (LTE4) and 11-dehydro-TXB2 levels are often used to assess cysteinyl LT and TXA2 production in humans. To define the potential role in the pathogenesis of asthma, we investigated the urinary LTE4 and 11-dehydro-TXB2 levels. LTE4 and 11-dehydro TXB2 levels were determined using liquid chromatography/tandem mass spectrometry (LC/MS) and gas chromatography/mass spectrometry (GC/MS), respectively. Urinary LTE4 levels in asthmatic patients (192 +/- 122 pg/mg creatinine, n = 14) were significantly higher (P < 0.005) than those in healthy volunteers (55 +/- 16 pg/mg creatinine, n = 13), but no significant difference in 11-dehydro-TXB2 levels was observed. A significant inverse correlation (r = -0.821, P < 0.005) was found between urinary LTE4 levels and the forced expiratory volume in 1 s (FEV1) but no significant correlation was observed between urinary 11-dehydro TXB2 levels and FEV1. The present findings suggest that cysteinyl LTs play a more important role in the pathogenesis of asthma than TXA2. PMID- 11060903 TI - The heart of the matter: R. D. Laing's enigmatic relationship with psychoanalysis. PMID- 11060904 TI - On R. D. Laing: style, sorcery, alienation. PMID- 11060905 TI - Awakening to love: R. D. Laing's phenomenological therapy. PMID- 11060906 TI - R.D. Laing's contribution to existentialism and humanistic psychology. PMID- 11060907 TI - R. D. Laing's contribution to the "treatment" of "schizophrenia": responsible responses to suffering and malaise. PMID- 11060908 TI - R. D. Laing's existential-humanistic practice: what was he actually doing? PMID- 11060909 TI - Improving anticoagulant care in Scotland--can we "go Dutch"? PMID- 11060910 TI - Intracerebral haemorrhages and oral anticoagulation in the north of Scotland. AB - The aim of this study has been twofold: 1--to examine the impact of oral anticoagulant (OAC) use on a possible recent rise in the admission rate of intracerebral haemorrhages to Aberdeen Royal Infirmary (ARI), and 2--to estimate the absolute risk of intracranial haemorrhage for outpatients followed up in the OAC Clinic at ARI. The number of patients admitted to ARI with intracerebral bleedings increased by 60% between 1993 and 1998. A corresponding increase in the proportion of patients with concurrent OAC use (4.7% vs 15.7%, p = 0.055) cannot sufficiently explain the increase in the total number of intracerebral haemorrhages. The average annual incidence of intracranial haemorrhages for the OAC Clinic at ARI is found to be acceptably low at 0.33% per year. Further audit of the large number of patients receiving warfarin outwith the supervision of the clinic is urgently required. PMID- 11060911 TI - Course, management and outcome of oral-anticoagulant-related intracranial haemorrhages. AB - The aim of this study was to compare the clinical course and radiological features of oral anticoagulant (OAC)-related intracranial haemorrhages with those of haemorrhages unrelated to OAC use admitted over the last six years to a tertiary care centre in the North of Scotland. We furthermore wished to determine the measures taken for reversal of OAC therapy and the resulting short-term outcome. Sixty-eight patients had been treated with OACs at the time of intracranial haemorrhage (32% subdural, 62% intracerebral). Patients admitted with OAC-related and unrelated haemorrhages did not differ significantly in any of the clinical features considered. On CT scan, there was no significant difference according to OAC use in the mean size of subdural (depth 15 +/- 5 vs. 18 +/- 8 mm, p = 0.36), or intracerebral haematomas (max. diameter 40 +/- 21 vs. 41 +/- 20 mm, p = 0.73). No reversal measures were taken in 38% of OAC-treated patients. In-hospital mortality was significantly higher for OAC-related haemorrhages compared to unrelated haemorrhages (38% vs. 18%, p = 0.001). To further elucidate the effects of anticoagulant reversal on the outcome of OAC related intracranial haemorrhages, a large-scale prospective study is warranted. PMID- 11060912 TI - Which acute stroke patients with atrial fibrillation are prescribed warfarin therapy? Results from one-year's experience in Dundee. AB - The recommended treatment of ischaemic stroke patients with atrial fibrillation (AF) is anticoagulation therapy with warfarin sodium and if this is contraindicated then aspirin should be used. The management of patients on warfarin therapy can be complicated and there is a risk of intra-cranial haemorrhage in elderly patients. However, these are the patients who stand to gain the most benefit from this treatment and therefore increased use of warfarin for secondary prophylaxis is likely to lead to a lower rate of subsequent admissions and less morbidity. The recommended treatment for these patients has often not been fully instigated in practice. This study was carried out in order to determine whether a group of patients admitted to a teaching hospital with diagnosis of ischaemic stroke and atrial fibrillation received appropriate antithrombotic therapy. Details of patients admitted with acute stroke during 1997 were obtained from the Dundee Stroke Database and information was extracted from the relevant clinical notes. Twenty-five out of 42 patients (60%) were considered eligible for anticoagulation and 14 out of those 25 (56%) were found to be on warfarin either on admission or subsequently. Of patients aged less than 75 years, 8/10 (80%) were on warfarin, whereas only 6/15 (40%) of those aged 75 years and older were being anticoagulated. PMID- 11060913 TI - The incidence of neurosurgical disease in patients who are referred to a neurosurgical centre for CT scanning: should district general hospital scanners be more active? AB - This study assessed the incidence of neurosurgical intervention in those patients who were referred to a neurosurgical unit for a CT scan in whom the diagnosis was unclear. A prospective consecutive patient survey was performed over a 29 week period in a regional neurosurgical unit. One hundred patients were included. Twenty-eight patients required neurosurgical management, of whom, nine required surgery within 24 hours. Twenty-three patients required ventilation, of whom 14 did not have a neurosurgical condition. In this group of patients, the majority did not have a neurosurgical condition. Increasing the availability of scanning services in district hospitals may avoid the unnecessary transfer of potentially unstable patients. PMID- 11060914 TI - Mental health in medical students. A case control study using the 60 item General Health Questionnaire. AB - This paper describes a cross-sectional case control study to measure the prevalence of psychological morbidity in first year medical students and compare it to the prevalence in in a randomly selected control group of other first year students at Edinburgh University. The study was conducted anonymously using the 60 item General Health Questionnaire. Participation rates were over 90% in both subjects and controls. A total of 17% of medical students had symptoms of psychological morbidity which may benefit from treatment and a further 29% of medical students had symptoms of psychological distress which would be expected to remit spontaneously. A similar rate was found in the control group of students. This suggests that if medical students or doctors, later in their careers, fare badly in terms of mental health then this may well be related to aspects of their lives and is not an intrinsic characteristic. PMID- 11060915 TI - The importance of identifying intracranial haemorrhage as a cause of transient focal neurological symptoms. AB - Until recently, there have been no reports of intracerebral haemorrhage presenting with transient neurological symptoms. We present two cases of intracerebral haemorrhage presenting as transient ischaemic attacks and discuss the radiological changes on early and late CT scans. It would seem justified to scan most patients presenting with TIA early to institute appropriate secondary prevention measures. PMID- 11060916 TI - Vertebral artery dissection diagnosed by non-invasive magnetic resonance imaging. AB - A forty-year-old man developed right-sided neck discomfort whilst cycling to work. On admission to hospital he was found to have signs of bilateral cerebellar dysfunction. Magnetic resonance imaging of the brain demonstrated bilateral areas of cerebellar infarction. Doppler ultrasound of the vertebral arteries was abnormal and non-invasive gradient echo time of flight magnetic resonance angiography confirmed the clinical diagnosis of vertebral artery dissection. The patient was anticoagulated for a period of three months and made a full recovery. PMID- 11060917 TI - An audit of the provision of parenteral nutrition in two acute hospitals: team versus non-team. AB - Studies have indicated that provision of artificial nutritional support services by a multidisciplinary nutrition support team results in significant health benefits and cost savings. An audit was conducted to compare the provision of parenteral nutrition in two hospitals, one with a nutrition support team and one without, with published standards for nutritional care. In the hospital with the nutrition support team there was greater use of nutritional assessment, the energy content of prescribed regimens and energy intake of patients was closer to estimated requirements and the incidence of some metabolic complications was lower. It has been recommended that a nutrition support team should be formed in the hospital that currently has no such facility. PMID- 11060918 TI - Letter from the editor. The computer age. PMID- 11060919 TI - The case for colorectal cancer screening. PMID- 11060920 TI - Accuracy of double-contrast barium enema in diagnosing colorectal polyps and cancer. AB - CRC is a common malignancy, and reduced mortality can be achieved through detection and treatment of early cancers and by removal of colonic adenomas. Although current screening recommendations, especially in the average-risk individual, typically promote the use of FOBT and FS, a substantial minority of colonic cancers and many colonic adenomas are not detected by these methods. Modalities that examine the entire colon, such as the barium enema and colonoscopy, can detect most clinically important colorectal neoplasms; however, their additional costs and potential risks have limited their use as initial screening examinations. But recent changes in governmental policies regarding reimbursement for CRC screening and increasing emphasis on total colon examinations have altered these recommendations. This review on the accuracy of the DCBE has emphasized the detection of colonic polyps and cancers and has updated the changing role of this examination in screening patients at variable risk for CRC. The efficacy of the barium enema depends on many factors that radiologists must understand and control to perform accurate examinations. Current recommendations for CRC screening and approved reimbursement of the barium enema for that purpose provide a new impetus to radiologists to maintain and improve their skills in performing and interpreting this radiologic examination. The barium enema may have a future in the new millennium. PMID- 11060921 TI - Tips for the comfortable double-contrast barium enema: the open tube technique with active drainage. PMID- 11060922 TI - Diagnosis of colorectal neoplasms on double-contrast barium enemas: interpretive aspects. PMID- 11060923 TI - Staging of colorectal cancer. AB - CT, MR, and TRUS play complementary roles in staging CRC. Further improvements in these techniques will improve the accuracy of preoperative staging and thereby help optimize patient treatment and outcome. PMID- 11060924 TI - Computed tomographic colonography: current and future status for colorectal cancer screening. PMID- 11060925 TI - The role of MR colonography for colorectal cancer screening. PMID- 11060926 TI - The role of colonoscopy for screening of colorectal cancer. PMID- 11060927 TI - [Clinical neosporosis in the dog: a review]. AB - Neospora caninum is an intracellular protozoan parasite that was first recognized in dogs in 1988. N. caninum may cause neuromuscular disease in dogs. Later, it was discovered that N. caninum has a wide host range and is an important cause of abortion in cattle. In this article, the literature on N. caninum in the dog is reviewed, with emphasis on clinical signs, pathology, diagnosis, treatment, and prognosis. PMID- 11060928 TI - [The role of the dog in the epidemiology of neosporosis in cattle]. AB - Neospora caninum is an intracellular protozoan parasite that was discovered in a dog in 1988. Since then, N. caninum has been demonstrated in a variety of animal species and it has been recognized as an important cause of abortion in cattle. An infection with N. caninum can be maintained in cattle herds for several generations by transplacental transmission from cow to calf. Recently, it was demonstrated that dogs can act as definitive hosts of N. caninum and therefore may be a source of infection for other species by shedding oocysts. Further evidence of a role of the dog in spreading the infection to cattle has been derived from epidemiological studies. The present state of knowledge is reviewed in this paper. PMID- 11060929 TI - [Some misunderstandings about the angora rabbit]. PMID- 11060930 TI - [Discussion about cow comfort and claw health]. PMID- 11060931 TI - [About the KNMvD and the AUV]. PMID- 11060932 TI - [Verdict from the Veterinary Disciplinary Board concerning fraud]. PMID- 11060934 TI - [The cattle veterinarian as interdisciplinary advisor]. PMID- 11060933 TI - [On request from the editor: "Letter to a student friend"]. PMID- 11060935 TI - Counterconditioning in the treatment of spider phobia: effects on disgust, fear and valence. AB - From the perspective that disgust is a core feature of spider phobia, we investigated whether the treatment efficacy could be improved by adding a counterconditioning procedure. Women with a clinically diagnosed spider phobia (N = 34) were randomly assigned to the regular one-session exposure condition (EXP) or to the exposure with counterconditioning condition (CC). In the CC-condition tasty food-items were used during the regular exposure exercises and the participants' favourite music was played. Both treatment conditions appeared very effective in reducing avoidance behaviour and self-reported fear of spiders, strongly attenuated the disgusting properties of spiders and altered the affective evaluations in a positive direction. CC was not more effective in altering the affective valence of spiders than EXP and was not superior with respect to the long term treatment efficacy at 1 year follow up. Apparently, regular exposure treatment is already quite effective in altering the affective evaluative component of spider phobia and it remains to be seen whether it is possible to further improve treatment outcome by means of procedures which are specifically designed to reduce the spiders' negative affective valence. PMID- 11060936 TI - Mood, personality disorder symptoms and disability in obsessive compulsive hoarders: a comparison with clinical and nonclinical controls. AB - Hoarding is a symptom of obsessive compulsive disorder (OCD), as well as a diagnostic criterion for obsessive compulsive personality disorder (OCPD). One recent study suggests that people who suffer from compulsive hoarding report more general psychopathology than people who do not [Frost, R.O., Krause, M.S., & Steketee, G. (1996). Hoarding and obsessive compulsive symptoms. Behavior Modification, 20, 116-132]. The present study addressed whether persons with OCD hoarding exhibit more depression, anxiety, OCD and personality disorders symptoms than community controls, OCD nonhoarders, or other anxiety disorder patients. Disability was also examined. Hoarding subjects were older than the other three groups, but age did not account for any of the differences observed among the groups. Compared to controls, OCD hoarding, nonhoarding OCD and anxiety disorder patients showed elevated YBOCS scores, as well as higher scores on depression, anxiety, family and social disability. Compared to nonhoarding OCD and anxiety disorder patients, OCD hoarding patients scored higher on anxiety, depression, family and social disability. Hoarding subjects had greater personality disorder symptoms than controls. However, OCD hoarding subjects differed from OCD nonhoarding and anxiety disorder subjects only on dependent and schizotypal personality disorder symptoms. The findings suggest that hoarding is associated with significant comorbidity and impairment compared to nonhoarding OCD and other anxiety disorders. PMID- 11060937 TI - Prospective evaluation of the etiology of anxiety sensitivity: test of a scar model. AB - A large body of research has suggested that anxiety sensitivity (AS) acts as a specific vulnerability factor in the development of anxiety pathology. More recently, attention has turned to the etiology of AS per se. The present study represents a specific test of a Scar model of AS. A Scar model posits that the experience of distress will affect the vulnerability factor. We were specifically interested in evaluating the effects of a specific stressor (spontaneous panic) as well as general distress on changes in AS over time. A large nonclinical sample of young adults (N = 1296) was prospectively followed over a five week highly stressful period of time (i.e. military basic training). Findings were consistent with the Scar model and suggested that the specific stressor of experiencing a panic attack as well as general stressors creating significant anxiety symptoms uniquely contributed to increased levels of AS (regardless of prior history of panic). Moreover, the experience of spontaneous panic in the context of generally low levels of distress (both anxiety and depression) appeared to be particularly pernicious in terms of resulting in greater increases in AS. In sum, anxiety-related stressors appear to have the potential to 'scar' individuals in regard to this cognitive vulnerability factor. PMID- 11060938 TI - Intrusive thoughts and auditory hallucinations: a comparative study of intrusions in psychosis. AB - Several theories of auditory hallucinations implicate the involvement of intrusive thoughts and other theories suggest that the interpretation of voices determines the distress associated with them. This study tested the hypotheses that patients who experience auditory hallucinations will experience more intrusive thoughts and be more distressed by them and interpret them as more uncontrollable and unacceptable than the control groups. It also examines whether the interpretation of hallucinations is associated with the distress caused by them and whether there are differences in the way that patients respond to and interpret their thoughts and voices. A questionnaire examining the frequency of intrusive thoughts and the reactions to them was administered to a group of patients with a diagnosis of schizophrenia who experienced auditory hallucinations, a psychiatric control group and a non-patient control group. In addition, the patients in the first group completed a similar questionnaire in relation to their voices. Analyses of covariance showed that patients who experienced auditory hallucinations had more intrusive thoughts than the control groups and that they found their intrusive thoughts more distressing, uncontrollable and unacceptable than the control groups. Correlational analyses revealed that patients' interpretations of their voices were associated with the measures of distress in relation to them. Repeated measures analyses of covariance found no differences between thoughts and voices on the dimensions assessed. The theoretical and clinical implications of these findings are discussed. PMID- 11060939 TI - Probability ratings in claustrophobic patients and normal controls. AB - Forty-nine DSM-IV diagnosed claustrophobics and 49 sex- and age-matched community controls, without any current or past psychiatric disorder, were asked to estimate the probability that three types if events would occur if they were in the described situations. The events were claustrophobic, generally negative, and positive in nature. The results showed that claustrophobics significantly overestimated the probability of events they specifically feared, i.e. the claustrophobic events, while there was no difference between the groups regarding generally negative events and positive events. This finding remained when the higher scores for claustrophobics on the Claustrophobia scale and the Anxiety Sensitivity Index were covaried out. The conclusion that can be drawn is that claustrophobics' probability ratings are characterized by distortions that are specifically connected to anxiety-arousing events and not negative events in general. The hypothesis is proposed that this may be explained by an exaggerated use of simplified rules-of-thumb for probability estimations that build on availability in memory, simulation, and representativity. PMID- 11060940 TI - Suppressing and attending to pain-related thoughts in chronic pain patients. AB - Attempted suppression of pain-related thoughts was investigated in consecutive referrals for pain management (N = 39). Participants monitored their pain-related thoughts for three 5-min periods. In period 1, all participants were instructed to think about anything. For period 2, participants were instructed to either suppress pain-related thoughts, attend to pain-related thoughts, or to continue to think about anything. In period 3, all participants were again instructed to think about anything. Participants instructed to attend to their pain reported more pain-related thoughts than suppressors and controls in both periods 2 and 3. Suppressors experienced reduced pain-related thoughts during period 2. There was no immediate enhancement or delayed increase. PMID- 11060941 TI - The development and validation of Spanish versions of the State and Trait Food Cravings Questionnaires. AB - OBJECTIVE: We developed and tested the psychometric properties of Spanish versions of the Trait and State Food Cravings Questionnaires (FCQ-T and FCQ-S respectively). METHOD: The instruments were translated and adapted to Spanish and administered to undergraduate students from a Southern university in Spain (N = 271). The data were analyzed using confirmatory factor analysis to compare the factor structure of the English and Spanish versions of both questionnaires. RESULTS: The factors structure of both questionnaires obtained excellent fit indices across their Spanish versions with the one exception that some factors of the FCQ-S were more highly intercorrelated among the Spanish sample than the American. DISCUSSION: This study supports the conceptualization of food cravings as universal multidimensional motivational states that can be reliably measured and supports the use of the Spanish versions of the FCQ. PMID- 11060942 TI - Specialist lists. PMID- 11060943 TI - Xylotox labelling. PMID- 11060944 TI - CPD learning. PMID- 11060945 TI - Timings inquiry. PMID- 11060946 TI - Drinking to excess? PMID- 11060947 TI - Antibiotic use. PMID- 11060948 TI - Sign of the times. PMID- 11060949 TI - As others see us. AB - Another installment from the literary archives focuses on the public image of dentists and dentistry. The feedback is not exactly flattering. PMID- 11060950 TI - Patients with cardiac disease: considerations for the dental practitioner. AB - The provision of dental treatment under both local anaesthesia and sedation has an excellent safety record, although medical problems may occur. The high prevalence of cardiac disease in the population, particularly ischaemic heart disease, makes it the most common medical problem encountered in dental practice. Additionally, the increasing survival of children with congenital heart disease makes them a significant proportion of those attending for dental treatment. While most dental practitioners feel confident in performing cardio-pulmonary resuscitation, treating patients with co-existent cardio-vascular disease often causes concern over potential problems during treatment. This article aims to allay many of these fears by describing the commoner cardiac conditions and how they may affect dental treatment. It outlines prophylactic and remediable measures that may be taken to enable safe delivery of dental care. PMID- 11060951 TI - Dental post-operative sensitivity associated with a gallium-based restorative material. AB - INTRODUCTION: This study forms part of a 2-year longitudinal clinical trial to compare the performance of a gallium-based restorative material (Galloy) with a high copper, mercury based (Dispersalloy) control material. METHOD: Following Ethical Committee approval, 25 galloy restorations and 25 Dispersalloy controls were placed in 14 adult patients, by a single operator. The cavities were of moderate size, indicating the use of amalgam as the restorative material. All restorations were polished within 1 week of placement, photographed and a silicone impression of the tooth and restoration recorded. In addition, a visual analogue scale (VAS), indicating the extent of any post-operative sensitivity, was completed by each patient for each restoration, immediately prior to polishing. A score of 0 indicated no sensitivity, while a score of 10 indicated the greatest possible sensitivity. At 6-month recall, the VAS scores, silicone impressions and photographs were repeated. RESULTS: The mean sensitivity scores for the galloy and Dispersalloy restorations at 1 week were 5.1 (+/- 3.4) and 1.0 (+/- 1.5), respectively and at 6 months, 1.8 (+/- 3.0) and 0.2 (+/- 0.1) respectively. The differences between these means at 1 week and at 6 months were significant (P < 0.01). CONCLUSION: Galloy restorations were associated with a much greater severity of post-operative sensitivity than Dispersalloy restorations. PMID- 11060952 TI - A survey of alcohol and drug use among UK based dental undergraduates. AB - OBJECTIVE: This study was designed to investigate the prevalence of alcohol and drug use. DESIGN: Anonymous self-report questionnaire. SETTING: A UK dental school in May 1998. SUBJECTS AND METHODS: 1st-5th year dental undergraduates (n = 264) were questioned on their use of alcohol and tobacco, cannabis and other illicit drugs whilst at dental school, and before entry. RESULTS: Eighty two per cent of male and 90% of female undergraduates reported drinking alcohol. Of those drinking, 63% of males and 42% of females drank in excess of sensible weekly limits (14 units for females, 21 units for males), with 56% of males and 58.5% of females 'binge drinking'. Regular tobacco smoking (10 or more cigarettes a day) was found to have a statistically significant association with year of study, 4th 5th year undergraduates being eight times more likely to regularly smoke than their junior colleagues. Fifty five per cent of undergraduates reported cannabis use at least once or twice since starting dental school, with 8% of males and 6% of females reporting current regular use at least once a week. CONCLUSION: Dental undergraduates are drinking above sensible weekly limits of alcohol, binge drinking and indulging in illicit drug use. Dental Schools should designate a teacher responsible for education of undergraduates regarding alcohol and substance abuse. PMID- 11060953 TI - The size of occlusal rest seats prepared for removable partial dentures. AB - OBJECTIVE: The aim of this study was to test whether rest seats cut by a group of general dental practitioners for a removable partial denture differed in size and shape from those prepared by either a group of postgraduate students or their academic teachers. METHOD: The occlusal surfaces of a number of plastic teeth were scanned by a laser profilometer. Each tooth was then placed in a set of articulated phantom head dental arches. 30 dental practitioners, 16 postgraduates and 11 dental academics were asked to cut a rest seat preparation in the mesial marginal ridge suitable for the construction of a removable chromium cobalt partial denture. The tooth was removed from the models, rescanned, and this data converted to grey level images for measurement of the width, length and area of each rest seat. Depth was calculated as the difference between the pre and post preparation scanned profiles. RESULTS: There was a wide variation in the size of the individual rest seat preparations. There was no significant difference between the measured parameters from the images of the preparations made by academic staff and postgraduate students. The two sets of data were therefore combined. The length, width and area of the rest seats prepared by the staff and postgraduate group were significantly greater than those cut by the dental practitioner group. However, there was no significant difference in the depths measured. The outline form of the rests prepared by the dental practitioners was often round with sharply defined margins contrasting with the smooth triangular preparation the staff and postgraduates prepared. CONCLUSION: A 'refresher' in tooth modification for GDPs designing partial dentures would improve the long term success of the prosthesis. PMID- 11060954 TI - Review of competency-based education in dentistry. AB - Dental education was previously structured in such a way that students largely learned what teachers chose to teach them. The aim was to produce a dentist with prescribed packages of knowledge upon graduation. This traditional approach was mainly disciplined-based. Increasingly, the current trend is towards competency based education, which provides a sequence of defined learning experiences to students so that on graduation they may be considered as qualified beginners in dental practice. The adoption of competency-based learning requires the re assessment and revision of old curricula, but there remains some common ground between traditional and competency-based education. PMID- 11060955 TI - Can calcium and vitamin D supplementation adequately treat most patients with osteoporosis? PMID- 11060956 TI - Osteoporosis: which current treatments reduce fracture risk? AB - Randomized controlled trials showed that several antiosteoporosis drugs decrease the incidence of fractures, which is a better measure of efficacy than are changes in bone mineral density or serum markers of bone turnover. With effective agents available, physicians should make osteoporosis treatment a priority, especially for patients at high risk, such as those who have already had a fracture. PMID- 11060957 TI - Tricyclic antidepressant poisoning. AB - Tricyclic antidepressant poisoning causes predictable electrocardiographic abnormalities and can be lethal. Cardiac arrhythmias, hypotension, seizures, and coma are common. Sodium bicarbonate is still considered the treatment of choice for severe toxicity, although a variety of supportive measures may be taken. Hypertonic saline appears to be a promising alternative. A QRS interval longer than 100 ms appears to be a better predictor of serious complications than is an elevated serum tricyclic antidepressant level. Cardiovascular toxicity is classically manifested as ventricular dysrhythmias, hypotension, heart block, bradyarrhythmias, or asystole. Activated charcoal binds tricyclic antidepressants. Give 30 to 50 g orally or by nasogastric tube with or without a cathartic (sorbitol 0.5 g/kg or 30 g of magnesium sulfate). Sodium bicarbonate is indicated if the QRS duration is more than 100 ms or the terminal right-axis deviation is more than 120 degrees. The suggested dosage is 1 to 2 mEq/kg, repeated as needed. Tricyclic antidepressants are used not only for depression but also for chronic pain syndromes, obsessive-compulsive disorder, panic and phobic disorders, eating disorders, migraine prophylaxis, and peripheral neuropathies. PMID- 11060958 TI - The evolving role of hormone therapy in advanced prostate cancer. AB - Earlier diagnosis and treatment of prostate cancer has changed the face of late stage disease, and the use of mainstay hormonal therapies--orchiectomy, luteinizing hormone releasing-hormone analogs, and combined androgen ablation- are evolving rapidly. New approaches such as antiandrogen monotherapy and intermittent therapy are being evaluated. In addition, palliative treatments for patients with androgen-independent tumors have expanded. The most common clinical presentation of advanced prostate cancer is a rising prostate-specific antigen level following primary therapy (radical prostatectomy or radiotherapy or both). Due to the negative psychological implications of orchiectomy, many patients are opting for treatment with luteinizing hormone-releasing hormone analogs. Because studies of combined androgen ablation have not provided conclusive results, it is reasonable to forego antiandrogen therapy for patients who undergo bilateral orchiectomy. Management options for patients with androgen-independent prostate cancer are expanding and include antiandrogen removal, antiandrogen therapy, and glucocorticoids. PMID- 11060959 TI - Diastolic dysfunction and heart failure: causes and treatment options. AB - Diastolic dysfunction is the underlying problem in one third of patients with heart failure, but it is still not well understood. Carefully excluding other causes of heart failure and recognizing indicators of diastolic dysfunction on invasive and noninvasive tests are important in establishing the diagnosis and in guiding therapy. Left ventricular relaxation and stiffness and left atrial function are the most important factors acting together to maintain adequate cardiac output under normal filling pressure. Echocardiography is the most important tool for the diagnosis of diastolic heart dysfunction. It is portable, safe, and excludes other causes of heart failure, such as valvular disease. Diuretics can be used to reduce volume overload, but caution is advised, as aggressive diuresis decreases stroke volume more in diastolic dysfunction than in systolic dysfunction. PMID- 11060960 TI - How to help your patients lose weight: current therapy for obesity. AB - Obesity is epidemic and dangerous. Weight loss is difficult but worth the effort. Although new weight-loss drugs are available, there are no magic bullets: to lose weight and keep it off, people must eat less and exercise more. This article presents a practical approach on how physicians can help their patients lose weight through diet, behavior modification, and adjunctive pharmacologic therapy. An appropriate initial goal is to lose 5% to 10% of one's baseline weight over 3 to 6 months. Drug therapy should not be used in isolation, but it can be an adjunct to diet, exercise, and behavior modification if a patient is committed and able to make necessary changes in eating and activity, and if the patient has a BMI of 30 or higher or a BMI greater than 27 with weight-related comorbid conditions. Anorectic therapy is unlikely to succeed and should be stopped if the patient does not lose at least 4 lb in the first 4 weeks of therapy. Orlistat is unlikely to be of benefit if patients do not lose at least 3% of their baseline weight by 12 weeks. Because obesity is a chronic disease, drug treatment should be continued indefinitely. The physician and patient must understand the intention to treat long-term. The weight loss plan devised should improve upon previous plans: for example, implementing a regular, convenient exercise program that had not been included in the past, or offering pharmacotherapy. PMID- 11060962 TI - Gastroesophageal reflux disease (GERD) PMID- 11060961 TI - Diagnosis and treatment of gastroesophageal reflux disease in the elderly. AB - Gastroesophageal reflux disease poses special diagnostic and therapeutic challenges in the elderly. These patients may not report the classic symptoms of dysphagia, chest pain, and heartburn, and they are more likely to develop severe disease and complications such as esophageal ulceration and bleeding. Therapeutic options include lifestyle changes, medication, and surgery. Polypharmacy and changes in renal, hepatic, and gastrointestinal function can complicate treatment. Proton pump inhibitors can help optimize disease management. The most common primary presenting symptoms of GERD in the elderly are regurgitation, dysphagia, dyspepsia, vomiting, and noncardiac chest pain, rather than heartburn. Because the elderly commonly take multiple drugs for various comorbidities, drug interactions and treatment responses must be carefully assessed in this patient population. Nonpharmacologic measures may be helpful but often do not relieve nighttime GERD symptoms. PMID- 11060964 TI - Putting soy in your diet PMID- 11060963 TI - Incorporating soy protein into a low-fat, low-cholesterol diet. AB - The US Food and Drug Administration recommends including four servings of at least 6.25 g each (25 g/day) of soy protein into a diet low in saturated fat and cholesterol to reduce the risk of heart disease. Patients are more likely to comply with this dietary change if they have their physician's support. The author discusses how the clinician can help patients incorporate soy protein into a low-cholesterol, low-fat diet. A meta-analysis found that soy protein consumption achieved an average 9.3% decrease in total cholesterol, a 12.9% decrease in low-density lipoprotein (LDL) cholesterol, and a 10.5% decrease in triglycerides. Soy pills and supplements such as isoflavone are not recommended. The cholesterol-lowering benefit has only been observed when the intact soy protein is used. Soy milk can be used in place of milk in coffee or over breakfast cereal, as well as in milkshakes and other blended drinks. Soy milk can be substituted for milk in many recipes. PMID- 11060966 TI - Get ready for CDR Weekly online. PMID- 11060965 TI - Case control study links salad vegetables to national increase in multiresistant Salmonella typhimurium DT104. PMID- 11060967 TI - The response of Indian gastroenterologists to Helicobacter pylori. PMID- 11060968 TI - Epidemiology of Helicobacter pylori in India. PMID- 11060969 TI - Transmission routes of Helicobacter pylori. PMID- 11060970 TI - Diagnosis of Helicobacter pylori infection in primary and tertiary care centers. PMID- 11060971 TI - Eradication of Helicobacter pylori in nonulcer dyspepsia. PMID- 11060972 TI - Treatment of peptic ulcer disease in the Helicobacter pylori era. AB - All patients with gastroduodenal ulcer disease should be tested and treated for Helicobacter pylori whether the ulcer is active or in remission. The rapid urease test, being the most commonly available, is the test of choice in our country. It is prudent to do H. pylori serology in the case of bleeding ulcers. Physicians should choose the regimen with the greatest efficacy regardless of costs. Bazzoli's low-dose triple therapy seems to be an excellent choice from our experience. Outside study protocols, routine checking for ulcer healing and eradication of H. pylori is recommended only for complicated ulcers. However, symptomatic patients need to be re-investigated with endoscopy and rapid urease test. PMID- 11060973 TI - Recurrence of Helicobacter pylori infection after eradication. PMID- 11060974 TI - Drug resistance in Helicobacter pylori infection. PMID- 11060975 TI - Helicobacter pylori--practical management issues. PMID- 11060977 TI - Treatment of Helicobacter pylori in peptic ulcer disease PMID- 11060976 TI - Diagnosis of Helicobacter pylori infection: who should be tested with intention to treat? PMID- 11060978 TI - Recurrence of Helicobacter pylori infection after eradication PMID- 11060979 TI - Drug resistance in Helicobacter pylori infection PMID- 11060980 TI - How organizational factors contribute to innovations in service delivery. AB - Twelve staff members from three community centers providing services to people with mental retardation were interviewed about the implementation of a new approach to service delivery that utilized Essential Lifestyle Planning. Findings indicate that although the innovation proposed and the technical assistance provided were quite similar, the outcomes varied greatly across the three sites. By highlighting the importance of organizational factors, such as timing and agency supports, these findings provide direction for future training and technical assistance activities of those who promote change in service-delivery philosophy and methods. PMID- 11060981 TI - Later-life planning: promoting knowledge of options and choice-making. AB - The effectiveness of a person-centered later-life planning training program designed to teach older adults (N = 60) with mental retardation about later-life planning issues and increase their participation in choice-making was examined. Using quantitative data analyses, we assessed the impact of the program on intervention and control groups. Results indicated that the intervention group gained more knowledge of concepts in the curriculum and made more choices over time than did the control group. The wide variety of goals that participants set were examined through qualitative analyses; 87% of the participants met or partially met their goals. Information on the supports and barriers to meeting goals is provided. PMID- 11060982 TI - Substitute decision-making and personal control: implications for self determination. AB - Levels of personal control exercised by 76 adults with mental retardation were contrasted by substitute decision-making status. Individuals with no guardian or conservator exercised more personal control than did those with a conservator, who exerted more personal control than did participants with a guardian. Similar group differences in self-determination competencies were also observed. When self-determination competencies were controlled statistically, significant group differences in exercise of personal control remained. Restrictive substitute decision-making status, inappropriate to current competencies, may have constrained individuals' levels of personal control. Reviewing substitute decision-making status on a regular basis and limiting or removing guardianship/conservatorship when it is not appropriate, may enhance personal control. PMID- 11060983 TI - Strengthening parent-community member relations on agency boards: comparative case study. AB - Relations between parents of children with developmental disabilities and other community members on agency boards can be difficult to negotiate. In this comparative study of the boards of three community agencies, we examined the forces that influence the quality of parent-community member relations. The results suggest that when differences between groups are acknowledged and respected at the same time that similarities are recognized and valued, board operations are more likely to be successful and focused on providing effective services. The case descriptions included here suggest that an organization's ability to manage the tension between intergroup differences and similarities is influenced by (a) organizational history of intergroup relations, (b) group identification, and (c) organizational practices that bridge group differences. Action implications are discussed. PMID- 11060984 TI - Changing visions into reality. PMID- 11060985 TI - Lessons from the margins, narrating mental retardation: a review essay. PMID- 11060986 TI - Looking backward, looking forward: mental retardation and the question of equality in the new millennium. PMID- 11060987 TI - Eugenics, "Baby Doe," and Peter Singer: towards a more "perfect" society: a response. PMID- 11060988 TI - Parallels in size of residential settings and use of Medicaid-financed programs. PMID- 11060989 TI - Nonmarital childbearing in the United States, 1940-99. AB - OBJECTIVES: This report presents information on trends and variations in nonmarital childbearing in the United States and includes information on the factors that have contributed to the recent changes. Data are presented for 1940 99 with emphasis on the trends in the 1990's. METHODS: Data in this report are presented on a variety of measures of nonmarital childbearing, including numbers, rates, and percent of births to unmarried women. Most of the data is from the National Vital Statistics System. Additional data explaining the trends shown are from the National Survey of Family Growth, the U.S. Census Bureau, and other national surveys. Data are presented and interpreted principally in charts and related tables. RESULTS: After rising dramatically during the half century from 1940 to 1990, out-of-wedlock childbearing leveled off, or slowed its rate of increase, in the 1990's. Many factors contributing to the long-term changes as well as more recent trends are described. These include changes in marriage patterns, sexual activity, contraceptive use, and abortion. The experience of the United States is put into context with data on nonmarital childbearing for other industrialized countries. Possible future trends in out-of-wedlock births are considered in the context of current population and birth patterns. PMID- 11060990 TI - [Estimation of age from teeth using the racemization of aspartic acid (racemization method)]. AB - We report a method for estimation of age from teeth using the racemization of amino acids (racemization method). This method is based on the characteristics of the constant age-related increase in the amount of D-aspartic acid in dentin. We estimated age by measuring the ratio of D-aspartic acid to L-aspartic acid, i.e. the ratio of racemization ?ratio of D/L, ln[(1 + D/L)/(1 - D/L)]?. Because different D/L ratios have been obtained from different teeth in the same individuals and from different sites of dentin in the same tooth, we usually prepare bucco-lingual longitudinal sections at the central part of each tooth, and prepare samples of powdered whole dentin. This powder is then mixed and used to measure the D/L ratio in the dentin. To accurately estimate age from forensic specimens, we simultaneously measured the D/L ratios in more than four control teeth of the same type obtained from subjects of known age. Use of control teeth is necessary because it is sometimes difficult to maintain constant running conditions for gas chromatography to obtain reproducible values in different runs. Therefore, for every measurement, we determined an equation for calculating age from the D/L ratios of control teeth, and estimated the age of the specimen tooth by substituting in its D/L ratio. The most reliable results were obtained using samples of lower incisors or premolars, which are single-rooted teeth with a relatively small volume of dentin. Thus sampling of the dentin is easier than for other teeth. It is better to keep control teeth desiccated because racemization does not proceed readily under such conditions. The deviation from the actual age in the cases we examined was less than 3 years. Thus, racemization of amino acids can be used for accurate estimation of age from teeth. PMID- 11060991 TI - [Application of the PCR-APLP method to determine ABO genotypes in forensic samples]. AB - We carried out ABO genotyping of forensic samples by the amplified product length polymorphism (APLP) technique. We present two novel systems. One is termed as eight primers system, in which eight allele-specific primers are added into a single PCR reaction. Another is termed as six primers system. In both APLP systems, all alleles were clearly detected using DNA purified from forensic samples. In PCR amplification with direct addition of specimen, ABO genotyping was also possible to blood stain, seminal stain, blood, saliva and urine. Furthermore, ABO genotyping worked only to chimpanzee. This PCR-APLP method should be convepffnt and valuable for forensic practice. PMID- 11060992 TI - [Immunological identification of human hemoglobin--a practical system for detecting human bloodstains]. AB - Identification of human blood is very important in the practice of criminal investigation. Methods that are species-specific and highly sensitive usually require special laboratory equipment. To develop a method that is specific, sensitive, and convenient for use at the crime scene, we applied a sandwich hybridization method for human blood identification. The test kit, which uses anti-human hemoglobin (Hb) monoclonal antibody, showed high species specificity and could detect as little as 20 ng human Hb. Cross-reactivity was observed only to baboon. It was able to detect dilutions up to 5,000,000 times and to identify a 15.5-year-old human bloodstain. Because the method is rapid (2 minutes) and does not require any special equipment, it is considered useful for crime scene investigation. PMID- 11060993 TI - [Application of Q.E.D. and Alco-Screen test kits to measurements of ethanol in forensic samples]. AB - We have investigated the applicability of the Q.E.D. (Quantitative Ethanol Detector) and Aloco-Screen test kits for screening ethanol concentrations in forensic samples, such as hemolyzed/decomposed blood, urine and vitreous humor. Because both kits were based on enzymatic color reactions, direct application of the kits to hemoglobin-rich samples gave unsatisfactory results. The deproteinization of blood with trichloroacetic acid followed by membrane filtration overcame such problem. This procedure was also effective for pretreatment of urine and vitreous humor samples to suppress excessive color development in the Alco-Screen test. The ethanol concentrations in whole blood (n = 29), urine (n = 7) and vitreous humor (n = 6) samples determined by the Q.E.D. kit correlated well with those determined by gas chromatography; the correlation coefficients were 0.986, 0.975 and 0.993, respectively. Because of its high specificity and sensitivity to ethanol, Q.E.D. seems to be highly reliable for quantitative estimation of ethanol concentrations in forensic samples. Alco Screen also had high sensitivity, the specificity to ethanol was relatively low; the color reaction was also observed in the presence of acetone, n-propanol, toluene, methanol, ethylene glycol, methamphetamine, diazepam and dichrovos. Therefore, if forensic samples are analyzed by the Alco-Screen, it is essential to confirm the positive results using other analytical methods. PMID- 11060994 TI - [Aircraft crashes in sky sports. Report of two autopsy cases and review of the accidents during 1981 to 1997 in Japan]. AB - The authors report two forensic autopsy cases of pilots who died in glider and ultra-light plane crashes in Aso, Kumamoto and review sky sports accidents in Japan (1981-1997). In the glider crash, sharp abdominal pain due to gallstones in a 78-year-old pilot was a possible cause of the accident. In the ultra-light plane crash, unskillful control of the plane by a 38-year-old pilot was the cause of the accident. The incidence of sky sports accidents increased from 12 cases in 1981 to 62 cases in 1997. The mortality rate of the victims of the accidents is very high. Investigation of natural diseases in pilots as a cause of accidents and the mechanisms of fatal injuries will help to assess preventive measures against sky sports accidents. PMID- 11060995 TI - [A proposal of essentials for forensic pathological diagnosis of sudden infant death syndrome (SIDS)]. AB - There are many sudden unexpected infant death cases which are easily diagnosed as sudden infant death syndrome (SIDS) both with or without autopsy in Japan. A SIDS diagnosis may provide a cover for accidental or criminal death. SIDS should not be a convenient diagnostic box that shelters the cases of unexpected infant death which lack the necessary antemortem information to make the correct diagnosis. The authors consider that SIDS should be diagnosed according to the direction of the international definition of SIDS, and propose the following essentials for a forensic pathological diagnosis. 1) A thorough autopsy should be performed based on precise autopsy protocol, including not only histological observation, but also, if necessary, toxicological, bacteriological, viral and/or biochemical examinations. 2) The forensic pathologist should be provided with pertinent information regarding antemortem health status, past clinical history, social circumstances, death scene investigation, etc. In order to collect more precise information, the authors recommend using a questionnaire such as the example in this report to record information from the deceased's guardians. 3) Suspicion of accidental death or infanticide should be completely ruled out. SIDS should be diagnosed only after these three essentials have been satisfied. When there is even a slight suspicion of accidental death or infanticide, or when the forensic pathologist can not obtain pertinent information about the deceased, the causes and classification of the death should be diagnosed as unspecified or undetermined. That is, the causes and classification of the death are undetermined as to whether it is a natural or unnatural death. Furthermore, several warning flags indicating a possible SIDS diagnosis were proposed: a case found dead in a supine position, the existence of a foreign body in the respiratory tract or mild infectious findings. The authors also emphasize the physician's responsibility to report a case found dead or dying of unnatural or clinically unexplained causes to the police. This is the crucial first step in getting an accurate diagnosis of SIDS. PMID- 11060996 TI - Role of calcium in bone health during childhood. AB - A discussion of observational and longitudinal studies examining the effect of early-life calcium intake on bone health is provided. A critical analysis of pediatric calcium supplementation trials is conducted by determining annualized percent changes in bone mineral density (BMD). The focus of the analysis is to identify consistent findings at specific bone sites, determine whether effects differed by the age of children studied, and establish the relationship between bone changes and baseline calcium intake. We found that increases in BMD owing to higher calcium intake among children appear to occur primarily in cortical bone sites, are most apparent among populations with low baseline calcium intakes, and do not seem to persist beyond the calcium supplementation period. Older (e.g., pubertal) children appear to have greater annual increases in lumbar BMD than younger (e.g., prepubertal) children. The annual percent increase in midradius BMD appears to be greater at higher intakes among the older children, but such a relationship is less apparent among the younger children. PMID- 11060997 TI - Studies on young child malnutrition in Iraq: problems and insights, 1990-1999. AB - Many reports on Iraq proclaimed a rise in rates of death and disease since the Gulf War of January/February 1991. Several of the studies on nutritional status are not readily accessible, and few have been compared to identify secular trends. Here, 27 studies examining nutrition among Iraqi children in the 1990s are reviewed. Only five studies were found to be of comparable methodologic quality. These are analyzed to identify major trends in child nutrition between August 1991 and June 1999. Limitations of existing studies and recommendations for future studies are discussed. PMID- 11060998 TI - Vitamin E and prevention of heart disease in high-risk patients. AB - Several observational studies have suggested that high intake of vitamin E may slow the development and progression of atherosclerosis. Some clinical trials also reported beneficial effects of vitamin E supplementation in the secondary prevention of cardiovascular events. However, results of recent large, multicenter clinical trials reported that vitamin E supplementation was not effective in reducing the incidence of cardiovascular events in high-risk patients. PMID- 11060999 TI - Evidence base for specialized nutrition support. AB - Data regarding the use of both parenteral and enteral specialized nutritional support (SNS) are available for a variety of common clinical scenarios. Herein, the data are reviewed for SNS in the context of critical illness, perioperative care, wasting syndromes (including HIV disease and cancer), and gastrointestinal disease (including short bowel syndrome, inflammatory bowel disease, and pancreatitis). PMID- 11061000 TI - Haemophilus influenzae type b vaccine in Papua New Guinea: what can we expect, and how should we determine priority for child health interventions? PMID- 11061001 TI - Salinity tolerance of Anopheles farauti Laveran sensu stricto. AB - To assess the salt tolerance of the malaria vector Anopheles farauti sensu stricto, larvae were collected from a freshwater environment on the outskirts of Honiara, Solomon Islands and placed in trays containing water with salinity varying from freshwater to seawater. Dead larvae and pupae and emerged adults were recorded and preserved. Most adults and nearly half of the larvae and pupae were then subjected to DNA analysis for species identification. No adult An. farauti emerged after prolonged immersion of larvae in undiluted seawater (3.5% salinity), although temporary immersion before pupation was compatible with survival. Salinities of up to 2.2% to 2.5% were compatible with good survival and adult emergence, at least from fourth instars. The results suggest that higher salinities may slow larval development and show that mortality at a given salinity is not uniform. PMID- 11061002 TI - Superior orbital fissure syndrome in a latent type 2 diabetic patient. AB - Although isolated cranial nerve palsies are common in diabetic patients, multiple, simultaneous cranial neuropathies are rare. We describe the second case of a complete superior orbital fissure syndrome including the optic nerve in a middle-aged Papuan man with newly diagnosed diabetes mellitus. The differential diagnosis included septic cavernous sinus thrombosis and Tolosa Hunt syndrome, and management was initially directed at excluding these serious, treatable conditions. PMID- 11061003 TI - Mortality rates and the utilization of health services during terminal illness in the Asaro Valley, Eastern Highlands Province, Papua New Guinea. AB - Between 1980 and 1989 we carried out fortnightly demographic surveillance in a random sample of people living in Goroka town, periurban areas and rural areas in the Lowa and Asaro Census Divisions, all within 1 1/2 hours' drive of the town in the Asaro Valley, Eastern Highlands Province. Cause of death was determined by verbal autopsy supplemented by any available health service information. Crude death and birth rates were 10 and 32 per 1000 person-years, respectively, in 59,906 person-years at risk. The standardized mortality ratio increased with increasing distance from town. Life expectancy at birth was 57 years for males and 55 years for females. The stillbirth rate was 19 per 1000 births, neonatal and infant mortality 21 and 60 per 1000 livebirths, respectively, and 1-4-year mortality 9 per 1000 person-years. Maternal mortality was 3 per 1000 births. Neonatal and infant mortality were respectively 7 and 3 times as high in Asaro Census Division as in Goroka town. Acute lower respiratory tract infections accounted for 22% of all deaths, chronic obstructive lung disease 10%, trauma 8% and gastroenteritis/dysentery 7%. 76% of deaths occurred at home and 44% of people who died had no treatment during their terminal illness. Health services were used most frequently by urban dwellers and by the young. To reduce mortality, a political commitment to provide functioning health services in rural areas is needed; regular supervision of health staff, ensuring the safety of staff and their families, availability of antibiotics as near people's homes as possible and regular mobile maternal and child health clinics are essential. Health education should include recognition of signs of severe disease and the importance of seeking treatment early. In view of high maternal and neonatal mortality, user fees should be waived for pregnant women. PMID- 11061004 TI - A familial cluster of Parkinson's disease identified in Milne Bay Province, Papua New Guinea. AB - Parkinson's disease is a chronic debilitating condition, the prevalence of which has not been fully established in Papua New Guinea. We describe a cluster of 9 cases of the disease, restricted to two generations of one family, and the key ideas and beliefs held within the family regarding disease aetiology. Many of the concerns and feelings of guilt expressed by family members were alleviated following supportive listening and culturally appropriate counselling, explanation and advice from trained health professionals assisted by bilingual family facilitators. This is the first time that such a family has been reported in Papua New Guinea and may warrant more detailed assessment. Addressing patient and community perceptions of disease aetiology should be at the heart of health promotion initiatives and counselling. PMID- 11061005 TI - Past and present research activities of the Papua New Guinea Institute of Medical Research. PMID- 11061006 TI - [Principles in construction of mathematical model for neonatal corpse cooling studies]. AB - The process of cooling of the body of a dead human was studied with consideration for the anatomical characteristics of adult and newborn human corpses. The body was simulated as a homogeneous elongated rotation ellipsoid and mathematical model was developed as function of relationship between body temperature and time. After appropriate characteristics in the mathematical model were established, cooling processes and effects of various factors on this process were analyzed for the final ellipsoid. The model suggests the possibility of amendments for environmental temperature and body parameters. PMID- 11061007 TI - [Forensic medical diagnosis of asymptomatic gastrointestinal disease]. AB - Twelve cases with congenital and acquired gastrointestinal diseases from expert practice are analyzed. The disease ran asymptomatically during life and eventuated in death. Potentialities of forensic medical diagnosis for such cases are demonstrated. PMID- 11061008 TI - [Forensic medical evaluation of professional working capacity in victims, who needs additional care]. AB - The philosophy of evaluating the need of a victim in extra care is discussed. The method for evaluating the need in transport vehicles for the victim is described in detail. Legislative documents which help solve such problems are cited, including those used by committees of forensic medical evaluations. PMID- 11061009 TI - [Forensic medical evaluation of harm severity inflicted to one's health by slight craniocerebral injuries in the remote period]. AB - A pressing problem is discussed: forensic medical evaluation of harm inflicted to health by slight craniocerebral injuries. Analysis of 2150 expert forensic medical and medico-social conclusions revealed serious organization and methodological errors in evaluation of harm inflicted to health by slight craniocerebral injuries. Approaches to improving the quality of such forensic medical expert evaluations are outlined. PMID- 11061010 TI - [Problems in organization of medical criminological registration and personality identification for subjects occupationally exposed to life risk]. AB - The paper discusses problems in organization of identification studies under conditions of mass deaths as exemplified by forensic medical records of medical criminological identification studies of subjects killed during war conflict in Chechnya. The evolution of the organization model of identification studies is shown transformation of organization philosophy, formation of expert algorithms, formalization and technologic realization of expert solutions. PMID- 11061011 TI - [Computer-aided personality identification by skull and life-time photography by POSKID 1.1 method]. AB - An improved method for computer-aided personality identification by the skull, based on the POSKID 1.1 software, consists in investigation of enlarged images of the skull and life-time photograph of the probable individual by coordinates of 49 anatomical points; independent quantitative evaluation of the aspect of each of the compared objects by the X, Y, and Z axes; formal evaluation of the results of comparative study of the skull-portrait by multidimensional discriminant analysis models. The proposed version differs from the POSKID 1.0 software in the method for evaluating the spatial position of the head on the portrait and adequate orientation of the skull in space, which necessitates the utilization of coordinate-regulated holder POSKID 1.1 method is based on multidimensional discriminant analysis and suggests a virtually reliable solution in 76.13-80.65% cases, a probable solution (positive and negative) in 11.61-18.06% cases, and motivated refusal from solution in 5.81-7.74% cases. In case of a probable or indefinite solution further investigations are recommended making use of life time photographs with different aspects. PMID- 11061012 TI - [Medical criminological personality identification by morphological signs of upper limbs segments]. AB - Studies of the morphology of segments of human upper limbs helped develop a method for medical criminological personality identification by metric parameters of the arm and hand in expert evaluation of a dismembered corpse. The sampling consisted of 270 corpses of Europeoids of both sexes (150 men and 120 women) aged 17-98 years. Ten quantitative signs of the upper limbs were analyzed. Statistical modeling of body length and weight and of head and chest circumferences was performed on the basis of the hand, arm, and forearm characteristics. The data can be used for rapid personality identification in criminal cases, war actions, terroristic acts, and calamities with multiple victims. PMID- 11061013 TI - [Typing of STR locus in biological objects with pronounced degeneration: analysis of DNA in exhumed remains of victims of war actions]. AB - The authors describe some specific features of enzymatic amplification typing of DNA preparations obtained from degraded tissues of remains of humans, which were brought from regions of war actions in the Chechen Republic. Special attention is paid to the specific artefact of polymerase chain reaction, for the first time detected by the authors in examinations of the above-mentioned objects. This so called "ladder" effect manifesting by simultaneous nonspecific amplification of many variants of allele fragments of the STR locus on the individual DNA matrix, which can be erroneously interpreted as an evidence of mixed DNA preparation, is apparently characteristic of individual objects with pronounced degradation of biological material. Such phenomena were observed in typing of STR locuses in biological tissues subjected to biological, thermal, and physicohemical degradation. The phenomenon was studied in detail. PMID- 11061014 TI - [Detection of P1 antigen in blood stains by absorption elution test with protease C]. AB - Some lots of goat anti-P1 sera intended for analysis of liquid blood can detect the antigen in blood stains by the absorption-elution test. PMID- 11061015 TI - [Study of microinjuries to hair cuticle of human head by scanning electron microscopy]. AB - The author presents the data on microclues in the free edges of hair cuticles of human head: microinjuries, microdefects (cracks, breaks, clefts, large and small waves) in representatives of Asia, Africa, Latin America, and Eastern Europe. The studies were carried out by scanning electron microscopy. Microinjuries of cuticle cells forming during hair combing by different types of combs and as a result of "permanent" curling are described. Scanning electron microscopy at the optimal magnification of x 3000 is a highly informative method for investigating microinjuries to the hair cuticle as additional signs in expert evaluation of the hair. PMID- 11061016 TI - [Method for estimating index of coronary arteries patency]. AB - The authors present an original method for evaluating the index of coronary artery patency by perfusion technology. The method is simple, requires no expensive equipment, and effectively diagnoses stenotic processes in coronary arteries. It was tried in cases of sudden cardiac death. The method is intended for forensic medical expert evaluations and for pathology studies. PMID- 11061017 TI - Medicalize rather than legalize street drugs. PMID- 11061018 TI - Risk issues associated with complementary and alternative medicine. PMID- 11061019 TI - Some emerging liabilities in complementary and alternative medicine. PMID- 11061020 TI - The Medical Examining Board and alternative medicine. PMID- 11061021 TI - Sudden cardiac death in a patient taking an over-the-counter diet pill. AB - The case of an otherwise healthy woman with a history of palpitations who survived sudden cardiac death from a ventricular arrhythmia is presented. Her only medications at the time of the event were alprazolan prn and over-the counter diet pills. The relation between ventricular arrhythmias and over-the counter diet pills is explored. Basic electrophysiologic principles are discussed as well as an overview for the work-up and long term management of sudden cardiac death. PMID- 11061022 TI - Reye's syndrome in an adult: a case report. AB - Reye's syndrome (RS), a condition characterized by encephalopathy and fatty liver, is associated with aspirin use and carries a poor prognosis. The majority of patients with RS are children and adolescents. We report a case of RS in an adult. PMID- 11061023 TI - A case of diarrhea and orthopnea in a 57-year-old female. AB - A 57-year-old female patient with known cardiac disease developed a 4 to 6 week history of diarrhea, followed by onset of orthopnea and subsequent right-sided cardiac failure. On hospital admission she was found to have pure tricuspid regurgitation, without evidence of cardiac ischemia, pulmonary embolism, bacterial endocarditis or pericardial disease. A 24-hour urine collection for 5 HIAA was elevated, and a subsequent octreotide scan documented abnormal uptake in the pelvic cul-de-sac. Bilateral ovarian masses were found at laparotomy, which on pathological examination were found to be a benign left ovarian cystic teratoma, and a right carcinoid tumor of the ovary. This patient presented with systemic complaints of diarrhea, and orthopnea and right sided heart failure that on evaluation were ultimately found to be due to a unilateral primary carcinoid tumor of the ovary, which accounts for less than 0.1% of all ovarian carcinomas, and only 5% of all carcinoids. Treatment of this malignant carcinoid syndrome presentation consisted of debulking of the tumor and continuation of her diuretics and digoxin. Diarrhea and orthopnea ceased within 2 weeks after her oophorectomy. On evaluation 6 weeks and 6 months postoperatively, her cardiac function was stable, though unchanged. 5-HIAA levels were within normal limits, demonstrating the curative function of surgery in patients with unilateral ovarian carcinoid without evidence of metastases, as well as preserved cardiac function in otherwise stable patients. PMID- 11061024 TI - The hepatopulmonary syndrome: a case report and review of the literature. PMID- 11061025 TI - Pseudoascites in the clinical setting: avoiding unwarranted and futile paracenteses. AB - Ascites is diagnosed on physical exam by findings of abdominal distension, bulging flanks, shifting dullness and a prominent fluid wave. However, as the following cases demonstrate, these signs may also be positive in pseudoascites due to thick layers of adipose tissue in the abdomen. A history of recent food binging and a lack of a prolonged prothrombin time should raise the index of suspicion for pseudoascites in a patient with a protuberant abdomen. In light of equivocal physical signs, physicians may employ ultrasonography to prevent patients with pseudoascites from suffering multiple futile attempts at paracentesis. PMID- 11061026 TI - Draft guidance on compliance plans for physician practices. PMID- 11061027 TI - Luminescence studies of the phagocyte response to endotoxin infusion into normal human subjects: multiple discriminant analysis of luminescence response and correlation with phagocyte morphologic changes and release of elastase. AB - A blood luminescence system (BLS) was employed to analyze blood phagocyte function in response to infusion of endotoxin (4 ng Escherichia coli lipopolysaccharide (LPS)/kg body weight) into 7 healthy human subjects. The subjects were closely monitored clinically, and extensive chemical, hematological and coagulation measurements were taken during the pretreatment, early (symptomatic, 1-8 h post-LPS), and late (asymptomatic, 12-48 h post-LPS) phases of acute inflammation. BLS assessment included measurement of basal and PMA stimulated phagocyte oxidase and myeloperoxidase (MPO) activities, and also included measurement of circulating (COR) and PAF-primed maximum (MOR) opsonin receptor-dependent phagocytic activities. Basal oxidase activity peaked at T + 1 h and showed an additional peak at T + 24 h post-LPS. The COR activity also peaked at 1-2 h, but remained elevated through T + 24 h post-LPS, while the basal MPO activity peaked only once at T + 1 h. We concluded that while MPO evidence of phagocyte respiratory activation returned to baseline by T + 4 h, COR evidence of receptor expression (receptor alert) remained elevated through T + 24 h. During this early (0-8 h) period, elastase/alpha 1AT complex concentration peaked at T + 3-4 h and again at T + 8 h. Peak numbers of circulating polarized and vacuolated phagocytes also appeared at T + 3 h and 7 h. We concluded that there was biochemical and morphological evidence of continuing phagocyte activity beyond T + 4 h to T + 8 h, and that this corresponded with the period during which the subjects were symptomatic. In addition, the appearance of a second peak of basal oxidase activity at T + 24 h, multiple discriminant analyses of all the luminescence data, and the sustained elevation of lactate suggested that there was a later second stage (T + 12 h to 48 h) of the human response to endotoxin, during which time the subjects were asymptomatic. PMID- 11061028 TI - Reversible binding of heparin to the loop peptide of endotoxin neutralizing protein. AB - Endotoxin neutralizing protein (ENP) from Limulus polyphemus is an amphipathic, 11.8 kDa protein with an isoelectric point of 10.2. ENP neutralizes lipopolysaccharide (LPS) and possesses antibacterial activity against Gram negative bacteria. Heparin binds to ENP and blocks its LPS-neutralizing activity. The relative blocking activity of heparin is equal to low molecular weight heparin and polyanetholsulfonic acid > heparan sulfate > chondroitin sulfate A > chondroitin sulfate C. Endoproteinase Glu-C hydrolysis of recombinant ENP results in four major peptides, three of which are seen following separation on reversed phase HPLC. Heparin binds to the loop peptide (31-72), which includes the heparin binding consensus sequence XBBXBX between the two cysteine residues of ENP. When heparin is added to the digest and then applied to a C18 column, the loop peptide is bound; however, it dissociates and elutes with either 5 M NaCl or 0.1 M sodium phosphate, demonstrating reversible binding to heparin. LPS and lipid A both bind to the loop peptide and remove it from digests of ENP; however, neither complex could be dissociated by salt or sodium phosphate. Heparin, LPS, and lipid A individually bind to the same site on ENP. PMID- 11061029 TI - A comparative trial of imipenem versus ceftazidime in the release of endotoxin and cytokine generation in patients with gram-negative urosepsis. Urosepsis Study Group. AB - Evidence from in vitro experiments and animal and human studies indicate that antibiotic therapy may induce the release of endotoxin from the outer membrane of Gram-negative bacteria. Antibiotics that bind preferentially to penicillin binding protein-2 (PBP-2)--such as imipenem--are associated with little release of endotoxin, while antibiotics that preferentially bind to PBP-3--such as ceftazidime--are associated with far greater release of endotoxin. We conducted a randomized, multicenter, double-blind study comparing imipenem to ceftazidime in patients with urinary tract infections caused by Gram-negative bacilli associated with signs and symptoms of systemic inflammation. A total of 33 patients were randomized to receive either imipenem (n = 14) or ceftazidime (n = 19) for initial treatment for urosepsis. No differences in plasma endotoxin, interleukin 6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) or urine endotoxin, IL-6 or IL 8 levels were found between the two treatment groups within the first 8 h after antibiotic administration. We conclude that, if differences exist with respect to endotoxin release by these two antimicrobial agents, these differences are not readily demonstrable in this clinical study with carefully defined patients with Gram-negative urinary tract infections. PMID- 11061030 TI - Lipopolysaccharide (LPS)-induced cell death of C3H mouse peritoneal macrophages in the presence of cycloheximide: different susceptibilities of C3H/HeN and C3H/HeJ mice macrophages. AB - Lipopolysaccharide (LPS) induced cytotoxicity toward mouse peritoneal macrophages from C3H/HeN mice but not C3H/HeJ mice in vitro in the presence of cycloheximide (CHX). More than 1 ng/ml LPS induced a significant time-dependent release of a cytoplasmic enzyme, lactate dehydrogenase (LDH), while even 1000 ng/ml LPS failed to induce it in LPS-non-responsive C3H/HeJ mouse macrophages. Although similar LPS-induced cytotoxicity was observed in a murine macrophage-like cell line, J774.1, but not in an LPS-resistant mutant of J774.1, the LPS1916 cell line, these results suggest that the induction of this cytotoxicity is linked to the LPS-sensitivity of mouse macrophages. A recombinant TNF-alpha (rTNF-alpha) at 100 ng/ml augmented LDH release from both C3H/HeN and C3H/HeJ macrophages treated with LPS and CHX, while rTNF-alpha alone or in combination with LPS or CHX failed to induce LDH release. These results suggest that this cytotoxicity might be partially regulated by high concentrations of exogenous TNF-alpha in both C3H/HeN and C3H/HeJ macrophages, implying a possibility of paracrine regulation of TNF alpha in mice toward LPS-treated macrophages under impaired protein synthesis. PMID- 11061031 TI - Endotoxin-induced alterations of lipid and fatty acid compositions in rat liver peroxisomes. AB - The structure/function of peroxisomal lipids in rat liver treated with a sublethal dose of endotoxin, a lipopolysaccharide (LPS), was investigated. Peroxisomes isolated from LPS-treated rat liver had remarkable alterations in lipid content compared with saline treated control liver peroxisomes. Cholesterol and phospholipids (PL) decreased significantly by 28.7% and 50.8%, respectively, leading to the change in the ratio of cholesterol/phospholipids (control 0.081 versus LPS 0.118, P < 0.001). A quantitative analysis from LPS-treated rat liver peroxisomes showed a general decrease in all classes of PL. No such alterations were observed in lipid content of other subcellular organelles. The peroxisomal fatty acid composition in LPS-treated animals was also altered. An analysis of fatty acid composition in PL and phosphatidylcholine from LPS-treated peroxisomes showed an increase in arachidonic acid (C20:4) and docosahexaenoic acid (C22:6). Very long chain (VLC) fatty acids (> C22:0) were also found increased in all classes of lipids in LPS-treated peroxisomes. Gadolinium chloride (GAD) mediated inactivation of Kupffer cells (KC) normalized cholesterol/PL ratio in LPS-treated peroxisomes. Collectively, the results indicate that the peroxisome metabolism of lipids and fatty acids is specifically altered in endotoxin-treated rat liver and at least part of the alterations may be mediated by factors released by KC. PMID- 11061032 TI - Limits of a deletion spanning Tlr4 in C57BL/10ScCr mice. AB - Proceeding from our observation that LPS-unresponsive mice of the strain C57BL/10ScCr mice fail to express the Tlr4 gene [Poltorak A, He X. Smirnova I et al. Defective LPS signaling in C3H/HeJ and C57BL/10ScCr mice: mutations in Tlr4 gene. Science 1998; 282: 2085], we have defined the exact limits of a deletion encompassing Tlr4 in the C57BL/10ScCr genome. The deletion removes 74723 bp of DNA, with reference to the control strain 129/J (from which the complete sequence of the Tlr4 locus was obtained). There is no inserted element, and no re arrangement of the chromosome (e.g. inversion or translocation) in the immediate region of Tlr4; the deletion removes only one recognizable gene. Hence, other immunological anomalies that have been identified in C57BL/10ScCr mice (a non healing phenotype in Leishmania inoculation and failure to produce interferon gamma in response to numerous microbial infections) must be ascribed to one of two causes. Mutation(s) at other loci may be responsible for these defects. Alternatively, Tlr4 locus deletion may have phenotypic consequences that exceed the well known blockade of LPS signal transduction. PMID- 11061033 TI - Synthesis of neoglycoproteins containing Kdo epitopes specific for Chlamydophila psittaci lipopolysaccharide. AB - The oligosaccharides alpha-Kdop-(2-->8)-alpha-Kdop-(2-->6)-beta-D- GlcpNAc-(1- >OAll) 4, alpha-Kdop-(2-->4)-alpha- Kdop-(2-->4)-alpha-Kdop-(2-->6)-beta-D GlcpNAc-(1-->OAll+ ++) 10, and the branched Kdo tetrasaccharide alpha- Kdop-(2- >4)-[alpha-Kdop-(2-->8)]-alpha-Kdop-(2-->4)-a lpha-Kdop-(2-->OAll) 21 have been prepared using en bloc transfer of Kdo oligosaccharide bromide donors to protected mono- or disaccharide acceptors. Radical addition of cysteamine to the anomeric allyl glycosides afforded good yields of the corresponding 3-(2 aminoethylthio)propyl glycosides 5, 11 and 22. The spacer ligands were activated with thiophosgene and reacted with bovine serum albumin to give the neoglycoconjugates 6, 12 and 23 which were used to prepare solid-phase antigens in enzyme immuno-assays for the characterization of monoclonal antibodies against chlamydial LPS. The data showed that the (2-->8)-linked Kdo disaccharide and the (2-->8)-(2-->4)-linked Kdo trisaccharide portion of the neoglycoconjugate 23 were not available for binding of antibodies which recognize these structures as di- and trisaccharide, respectively. PMID- 11061034 TI - Possible mechanisms of late-life-onset allergic diseases and asthma in the senior citizen. AB - The clinical spectrum of allergic diseases and asthma changes over the life span of the individual and is influenced by a wide variety of anatomic, physiologic, and immunologic factors. Nowhere do these changes play a more important role than in the elderly patient with allergic disease or asthma where the culmination of these events contribute to disease expression, which at times can result in irreversible endstage disease. It is estimated that between 2010 and 2030 the elderly population will increase by 75% and will represent a significant proportion of consumers of total health care resources. This presentation will examine possible mechanism(s) that contribute to the development of late-onset allergic diseases and asthma in the elderly as a possible basis for identification of antecedents of endstage disease and interventive strategies for the prevention of the irreversible consequences in this population. PMID- 11061035 TI - Immune-mediated dermatologic skin disorders in the senior patient. AB - The baby boomer population, marching on the journey from infancy through the portals of middle life, now enters an uncertain period of life in which the only certainty is the fact that all the biological clocks continue the countdown toward the inevitable failure of a previously well-served immune system. We are faced with the ever-increasing probability that the very immune system that sustained us so well throughout our youth will eventually falter, predisposing us to a vast array of disorders resulting in immune dysregulation. It is in this setting that I will review the dermatologic disorders that may befall the "senior" patient as we journey toward that stage of life where we, as physicians and as an aging population ourselves, encounter a whole new set of medical challenges as we make the turn at the bend in the road. Accordingly, this review will focus on autoimmune bullous disorders, cutaneous manifestations of systemic disease, endocrine disorders, skin cancers, and paraneoplastic syndromes. PMID- 11061036 TI - Ocular allergic disease in the senior patient: diagnosis and management. AB - Ocular allergic disease may be the most common clinical problem involving the eye. Ocular allergy encompasses a spectrum of diseases characterized by type I hypersensitivity responses, and knowledge of the pathogenesis of these diseases is central to understanding the role of therapeutic agents used to treat these diseases. The differential diagnosis of ocular allergic disorders includes any disease in which the eye becomes red associated with itching, burning, and tearing. Many therapeutic options are available; however, treatment strategies may require a combination of prescriptive drops to control the disease. PMID- 11061037 TI - Avoiding fraud and abuse in complying with the new federal documentation guidelines. PMID- 11061038 TI - Penicillin allergy: improving patient care and the medical record. PMID- 11061039 TI - The value of skin testing for penicillin allergy in an inpatient population: analysis of the subsequent patient management. AB - It was decided to assess the value of skin testing in a group of inpatients with a remote history of penicillin allergy, in terms of whether or not beta-lactams were subsequently given, if any adverse reactions occurred as a result of this therapy, and if labeling of the patient record was changed subsequent to skin testing and/or challenge. All patients seen in consultation with a history of penicillin allergy were assessed. When done, skin tests were performed with the major and minor determinants of penicillin and semisynthetic penicillins. Charts were reviewed after discharge in terms of the antibiotics given during admission, adverse events, and the medical record and hospital database labeling for drug allergy at discharge. Skin testing was carried out in 79% of 67 patients assessed and in all, the tests were negative. Beta-lactam therapy was recommended in 51/53 patients but was given in only 57% of these cases. At discharge, 49% of patients' records still carried the penicillin allergy label, despite negative skin testing and/or successful completion of a course of beta-lactam therapy. So, in approximately half of the patients reviewed, beta-lactams were not given despite negative skin tests and a recommendation to do so, if indicated, and 49% of patients were still inappropriately labeled as being penicillin-allergic on discharge. PMID- 11061040 TI - Latex protein: a hidden "food" allergen? AB - Avoidance of latex allergens is the primary method to prevent adverse reactions. Natural rubber latex is found in many different products in both the health care industry and in modern society, and consequently results in unexpected exposures of sensitized individuals. The use of latex gloves by food handlers provides one potential route for inadvertent exposure to latex allergens. In this study we have used two immunological methods to determine whether latex proteins are transferred to foods following contact with latex gloves. Direct transfer of latex protein to cheese was visualized using a modified immunoblot method. Sliced cheese was touched with a gloved finger. A nitrocellulose membrane was applied to lift the potential fingerprints and a rabbit anti-latex antiserum was used to visualize the transfer of any latex finger-prints. After handling lettuce with gloves, transferred protein was recovered by extracting the lettuce and quantified using an inhibition ELISA for latex proteins. Fingerprints of latex protein were readily detectable on cheese after contact with powdered latex gloves, but not with vinyl gloves. Furthermore, powdered latex glove use resulted in measurable amounts of latex protein on lettuce with an exposure-dependent increase in the latex protein levels. Lettuce alone or lettuce handled with vinyl gloves was negative for latex protein. The use of latex gloves by food handlers is the source of an indirect food additive in the form of latex proteins. It is recommended that food handlers avoid the use of latex gloves to eliminate inadvertent exposure of latex-sensitive individuals. PMID- 11061041 TI - Persistent cough: differential diagnosis. AB - Cough may be defined as a physiologic response to foreign or noxious substances that enter or irritate the respiratory tract. It is the fifth most common symptom complex for which patients seek medical care and which results in more than 30 million office visits per year. When cough is present for more than three weeks it is referred to as chronic or persistent cough. This presentation will examine the differential diagnosis of persistent cough together with a description of the autonomic innervation of the human airways, mechanism(s) of cough, and approach to the patient. PMID- 11061042 TI - Pathophysiology of bronchial inflammation: chemoreceptors as therapeutic targets. AB - Asthma is an inflammatory disease characterized by reversible airway obstruction from a combination of bronchoconstriction and inflammatory changes including airway edema, infiltration of inflammatory cells such as eosinophils, mast cells, CD4+ T helper cells and monocyte/macrophages. Pharmacotherapy approaches include relievers of acute symptoms and controllers of inflammation. Inhaled corticosteroids are the sentinel therapy for asthma inflammation but are not always totally effective for a variety of reasons. With the increased understanding about the molecular basis for asthma inflammation, new therapies are emerging to target specific molecular networks, including antibodies against IgE and the TH2 cytokine IL-5; soluble IL-4 receptors and recombinant TH1 cytokines such as IL-12. Further, allergen immunotherapy for asthma is based upon its ability to change a TH2 to a TH1 response by inhibiting IL-4 and/or stimulating IFN gamma production. Although each of these modalities have their potential strengths and weaknesses, the approach offers a fresh attempt to better define the syndrome called asthma, show diversity of etiologies within even the same patient, and develop more effective, long lasting therapies for patients with this condition. PMID- 11061043 TI - Combination of inhaled corticosteroids plus other medications in the management of moderate to severe persistent asthma. AB - Severe persistent asthma accounts for a small percentage, probably less than 5% of all patients with asthma, but is responsible for the major portion of health care costs associated with the illness. According to the National Institutes of Health (National Asthma Education and Prevention Program) guidelines for the management of asthma, patients with severe asthma should be treated with high dosages of inhaled corticosteroids. These inhaled corticosteroids can be given in conjunction with a brief course of oral or parenteral systemic steroids, but it is best to decrease or eliminate systemic corticosteroid therapy whenever possible to prevent the side effects of long-term oral prednisone therapy. If inhaled corticosteroids do not control the asthma, then one or perhaps two, and even three, other long-term control medications can be added to the therapy regimen. Current guidelines recommend adding a long-acting beta 2-agonist such as salmeterol to the inhaled corticosteroid. Recent evidence suggests that leukotriene receptor antagonists can also be used in conjunction with inhaled steroids. Theophylline is also recommended as another controller agent to be considered. Unfortunately, no studies have comparatively evaluated all of these different classes of agents, even in moderate asthma, in head-to-head trials. This manuscript will review the current literature and provide the author's perspective on the combination of these medications in the pharmacologic management of moderate to severe persistent asthma. PMID- 11061044 TI - Asthma among the famous. Djuna Barnes (1892-1982) American author. PMID- 11061045 TI - Brain natriuretic peptide increases acutely and much more prominently than atrial natriuretic peptide during coronary angioplasty. PMID- 11061047 TI - Detecting coronary artery calcification. PMID- 11061046 TI - True-positive exercise electrocardiogram/false-negative thallium-201 scintigram: a proposal of a mechanism for the paradox. PMID- 11061048 TI - The assessment of myocardial viability and hibernation using resting thallium imaging. AB - Rest-redistribution thallium-201 imaging is widely used to assess recovery of regional systolic dysfunction in patients with chronic coronary artery disease. In several studies, this technique has demonstrated very high sensitivity but reduced specificity, as reported in general for radionuclide imaging. In clinical terms, this implicates that many dysfunctional territories will not recover after revascularization despite a substantial amount of tracer uptake. Yet, the amount of tracer uptake in a given myocardial segment, although not perfect, remains the best indicator for predicting reversible dysfunction. In fact, the occurrence of redistribution after rest injection is not very common and it does not substantially contribute to the accuracy of the test. However, it is still undetermined whether the presence of redistribution is relevant for prognostic implications. PMID- 11061049 TI - New and emerging pharmacologic strategies in the management of chronic heart failure. AB - Chronic heart failure (CHF) is a complex syndrome involving activation of multiple cellular, metabolic, and neurohumoral pathways following the initial myocardial insult. Recently, there have been considerable advances in the pharmacologic management of CHF. The current approach to treatment recognizes the need to target neurohormonal activation, and the use of angiotensin-converting enzyme (ACE) inhibitors and beta blockers should now be regarded as part of standard therapy in many patients with CHF. However, because of the complexity of the disease, blockade of additional pathways is likely to be required to maximize the therapeutic benefit of intervention. To this end, there are several agents under active late-phase clinical evaluation. The most advanced of these new strategies (beyond renin-angiotensin-aldosterone blockade) is inhibition of the endothelin system. There is a substantial body of evidence that this system is intimately involved in CHF disease progression. Early-phase clinical data are very encouraging and support the potential utility of long-term endothelin inhibition. Other novel approaches involve the use of cytokine antagonists (e.g., agents that block tumor necrosis factor-alpha activity) and the augmentation of natriuretic peptides. If all these potential agents prove to be of benefit in CHF, the question of which agent or combination of agents to use in which patients will arise. There is therefore a need to develop scientific approaches in order to be able to identify more accurately patients who will obtain benefit from specific classes or combinations of drugs. PMID- 11061050 TI - Postoperative troponin I values: insult or injury? AB - BACKGROUND: Troponin I (TnI) is increasingly employed as a highly specific marker of acute myocardial ischemia. The value of this marker after cardiac surgery is unclear. HYPOTHESIS: The purpose of this study was to measure serum TnI levels prospectively at 1, 6, and 72 h after elective cardiac operations. In addition, TnI levels were measured from the shed mediastinal blood at 1 and 6 h postoperatively. Serum values were correlated with cross clamp time, type of operation, incidence of perioperative myocardial infarction, as assessed by postoperative electrocardiograms (ECG) and regional wall motion, as documented by intraoperative transesophageal echocardiography (TEE). METHODS: Sixty patients underwent the following types of surgery: coronary artery bypass graft (CABG) (n = 45), valve repair/replacement (n = 10), and combination valve and coronary surgery (n = 5). Myocardial protection consisted of moderate systemic hypothermia (30-32 degrees C), cold blood cardioplegia, and topical cooling for all patients. RESULTS: Of 60 patients, 57 (95%) had elevated TnI levels, consistent with myocardial injury, 1 h postoperatively. This incidence increased to 98% (59/60) at 6 h postoperatively. There was a positive correlation between the length of cross clamp time and initial postoperative serum TnI (r = 0.70). There was no difference in the serum TnI values whether or not surgery was for ischemic heart disease (CABG or CABG + valve versus valve). There were no postoperative myocardial infarctions as assessed by serial ECGs. There was no evidence of diminished regional wall motion by TEE. Levels of TnI in the mediastinal shed blood were greater than assay in 58% (35/60) of the patients at 1 h and in 88% (53/60) at 6 h postoperatively. Patients who received an autotransfusion of mediastinal shed blood (n = 22) had on average a 10-fold postoperative increase in serum TnI levels between 1 and 6 h. Patients who did not receive autotransfusion average less than doubled their TnI levels over the same interval. At 72 h, TnI levels were below the initial postoperative levels but still indicative of myocardial injury. CONCLUSION: Postoperative TnI levels are elevated after all types of cardiac surgery. There is a strong correlation between intraoperative ischemic time and postoperative TnI level. Further elevation of TnI is significantly enhanced by reinfusion of mediastinal shed blood. Despite these postoperative increases in TnI, there was no evidence of myocardial infarction by ECG or TEE. The postoperative TnI value is even less meaningful after autotransfusion of shed mediastinal blood. PMID- 11061051 TI - Aortic atherosclerotic lesions in the thoracic aorta detected by multiplane transesophageal echocardiography as a predictor of coronary artery disease in elderly patients. AB - BACKGROUND: The presence of atherosclerotic lesions in the thoracic aorta by transesophageal echocardiography (TEE) appears to be a marker for the presence of significant coronary artery disease (CAD) in the general population. HYPOTHESIS: We investigated whether atherosclerotic lesions in the thoracic aorta, by multiplane TEE, could be a marker for CAD in elderly patients. METHODS: In all, 127 patients (67 men, 60 women, aged 68 +/- 13 years), underwent a TEE study with imaging of the thoracic aorta and cardiac catheterization with coronary angiography. The presence of a distinct, linear, or focal, highly echogenic mass protruding into the vessel lumen was the criterion for the diagnosis of atherosclerotic plaque. RESULTS: Atherosclerotic lesions were found in 30 of 36 patients (83.3%) with and in 20 of 91 (22%) without CAD. Of the 41 patients > or = 70 years, atherosclerotic lesions were detected in 14 of 17 (82.3%) with and in 13 of 24 patients (54%) without CAD. The sensitivity, specificity, and positive and negative predictive values in this group were 82.3, 46, 52, and 78.6%, respectively. Multivariate logistic regression analysis revealed that in patients aged > or = 70 years only advanced atherosclerotic lesions were independent predictors of significant CAD. However, the high negative predictive value of the method indicates that the absence of aortic plaque is a strong predictor of the absence of CAD. CONCLUSIONS: The presence of atherosclerotic lesions in the thoracic aorta is a strong predictor of CAD only in patients < 70 years old. However, the negative predictive value of the method is high for all patients regardless of age. PMID- 11061052 TI - Emergent angioplasty prevents left ventricular dilation in patients with acute anterior wall myocardial infarction and cardiogenic shock. AB - BACKGROUND: Percutaneous transluminal coronary angioplasty (PTCA) reduces in hospital mortality and improves long-term outcome in patients with acute myocardial infarction (MI) complicated by cardiogenic shock. However, no study has evaluated the effects of different reperfusion therapies on left ventricular (LV) dimension and cardiac function in long-term survivors of MI with cardiogenic shock. HYPOTHESIS: We investigated the effects of PTCA on the development of LV dilation in patients who survived MI complicated by cardiogenic shock. METHODS: We studied 34 patients with a first MI and cardiogenic shock in whom two dimensional echocardiography was performed immediately after admission and 1 month after infarction. Group A consisted of 17 patients who underwent emergent PTCA during the acute phase of MI, and Group B consisted of 17 patients who did not undergo PTCA. We also studied 119 patients with a first uncomplicated acute anterior MI, including 53 who underwent PTCA (Group C) and 66 who did not (Group D). The length and wall thickness of the infarcted and noninfarcted endocardial segments were determined immediately after MI and 1 month later, and LV ejection fraction (LVEF) was measured during the chronic phase. RESULTS: The lengths of the infarcted and noninfarcted endocardial segments were significantly greater in Group B than in the other three groups (p < 0.05). The LVEF was significantly lower in Group B than in the other three groups (p < 0.05). CONCLUSIONS: We conclude that PTCA performed in patients during the acute phase of MI complicated by cardiogenic shock lowers in-hospital mortality and prevents both LV dilation and a decrease in LVEF. PMID- 11061053 TI - Improved survival after acute myocardial infarction in patients with advanced Killip class. AB - BACKGROUND: The continuing applicability of the Killip classification system to the effective stratification of long-term and short-term outcome in patients with acute myocardial infarction (MI) and its influence on treatment strategy calls for reanalysis in the setting of today's primary reperfusion treatments. HYPOTHESIS: Our study sought to test the hypothesis that Killip classification, established on admission in patients with acute MI, is an effective tool for early prediction of in-hospital mortality and long-term survival. METHODS: A series of 909 consecutive Olmsted County patients admitted with acute MI to St. Marys Hospital, Mayo Clinic, between January 1988 and March 1998 was analyzed. Killip classification was the primary variable. Endpoints were in-hospital death, major in-hospital complications, and post-hospital death. RESULTS: Patients analyzed included 714 classified as Killip I, 170 classified as Killip II/III, and 25 classified as Killip IV. Increases in in-hospital mortality and prevalence of in-hospital complications correspond significantly with advanced Killip class (p < 0.01), with in-hospital mortality 7% in class I, 17.6% in classes II/III, and 36% in class IV patients (p < 0.001). Killip classification was strongly associated with mode of therapy administered within 24 h of admission (p < 0.01). Killip IV patients underwent primary angioplasty most commonly and were less likely to receive medical therapy. CONCLUSIONS: Killip classification remains a strong independent predictor of in-hospital mortality and complications, and of long-term survival. Early primary angioplasty has contributed to a decrease in mortality in Killip IV patients, but effective adjunctive medical therapy is underutilized. PMID- 11061054 TI - Changes in QT dispersion during adenosine infusion. AB - BACKGROUND: QT dispersion (QTd) measurement during treadmill stress testing has been to shown to improve the accuracy of exercise electrocardiogram (ECG) in the detection of significant coronary artery disease (CAD). HYPOTHESIS: The aim of this study was to determine whether adenosine-induced changes in QTd could predict significant CAD and to assess its efficacy as a diagnostic index in patients undergoing adenosine stress test. METHODS: QT interval measurements were made in 57 consecutive patients undergoing adenosine sestamibi stress test. Patients with an abnormal stress test underwent coronary angiography. Patients with significant disease by coronary angiography (> 70% stenosis) were classified as having CAD (Group 1), and those with normal stress images and/or normal coronaries by angiography were classified as having no CAD (Group 2). RESULTS: QT dispersion increased from 28.2 +/- 4.5 to 43.8 +/- 4.5 ms with a delta QTd of 15.53 +/- 3.68 in Group 1 (p = 0.001) and from 28.4 +/- 2.6 to 34.8 +/- 2.8 ms with a delta QTd of 6.58 +/- 2.21 ms in Group 2 (p = 0.006). Patients in Group 1 had a significantly higher increase in QTd (delta QTd) than the patients in Group 2 (p < 0.03). Addition of delta QTd (> 10 ms) to the ST depression during adenosine infusion would increase the sensitivity of the ECG from 23 to 65% and decrease the specificity from 91 to 70% for diagnosis of significant CAD. CONCLUSIONS: delta QTd is significantly more prolonged in patients with CAD during adenosine infusion. It increases the sensitivity of the stress ECG in diagnosis of CAD during adenosine infusion when used as an adjuvant index. PMID- 11061055 TI - Monotherapy with amlodipine or atenolol versus their combination in stable angina pectoris. AB - BACKGROUND: The basic cause of angina pectoris is imbalance between the metabolic needs of the myocardium and the capacity of the coronary circulation to deliver sufficient oxygenated blood to satisfy these needs. HYPOTHESIS: The study was undertaken to evaluate whether the effect of combined amlodipine and atenolol therapy on patients with stable angina pectoris and with ST-depression during exercise testing and 48-h ambulatory electrocardiographic monitoring is superior to that of either agent given alone. METHODS: Patients with stable angina pectoris and ST depression during exercise and ambulatory monitoring were randomized to receive amlodipine (n = 116) or atenolol (n = 116), or both (n = 119). All patients were also treated with short- and long-acting nitrates. The design was a double-blind, randomized, triple-arm parallel group study with 10 weeks of administration of the test medication. RESULTS: In terms of time to onset of ST depression > 1 mm, time to onset of angina, total exercise time, maximum achieved workload, and peak intensity of angina, amlodipine and atenolol alone were as effective as their combination. During ambulatory monitoring, atenolol was more effective than amlodipine regarding total time and number of ST depression episodes, and as effective as the combined drugs. CONCLUSION: For individual patients with stable angina pectoris, combination of a beta blocker with a calcium antagonist is not necessarily more effective than either drug given alone. PMID- 11061056 TI - Relationship between mean right atrial pressure and Doppler parameters in patients with right ventricular infarction. AB - BACKGROUND: The incidence of an inferior left ventricular infarction involving the right ventricle is very high, ranging from 14 to 84%. Isolated right ventricular infarction accounts for < 3% of all cases of infarction. HYPOTHESIS: The aim of the present study was to assess the relationship between Doppler parameters of hepatic vein and tricuspid inflow, as well as mean right atrial (RA) pressure in patients with right ventricular infarction. METHODS: In all, 59 consecutive patients with inferior left ventricular infarction involving the right ventricle were selected for the study. All patients underwent Doppler echocardiographic evaluation of tricuspid and hepatic vein parameters and catheterization of the right side of the heart. Patients were divided into two groups according to the presence or absence of severe tricuspid regurgitation. RESULTS: In patients with severe tricuspid regurgitation, a significant correlation (r = 0.64; p < 0.001) between RA maximal volume and mean right atrial pressure (RAP) was found, and the sensitivity of RA maximal volume in identifying mean RAP > 7 mmHg was 64% with a specificity of 78%. In patients without severe tricuspid regurgitation, the most significant relationship was observed between mean RAP and inferior vena cava collapse index. Significant correlations between maximal and minimal diameters of the inferior vena cava were also observed. CONCLUSIONS: Echocardiographic and Doppler parameters may be useful for evaluating mean RAP in patients with right ventricular infarction. In patients with severe tricuspid regurgitation, the more important parameters are maximal and minimal RA volumes. In patients without severe tricuspid regurgitation together with right atrial volume, the important parameters are acceleration and deceleration time of the tricuspid inflow peak E velocity and hepatic systolic and diastolic venous flow. PMID- 11061057 TI - Transient increase in plasma brain (B-type) natriuretic peptide after percutaneous transluminal coronary angioplasty. AB - BACKGROUND: Brain (B-type) natriuretic peptide (BNP) is known to be secreted predominantly from the myocardium. Brain natriuretic peptide plasma concentrations have been shown to be markedly increased in patients with acute myocardial infarction; however, plasma BNP response during episodes of myocardial ischemia has not been established. HYPOTHESIS: This study was designed to examine plasma BNP in patients with transient myocardial ischemia induced by inflation of a percutaneous transluminal coronary angioplasty (PTCA) balloon. METHODS: Thirty consecutive patients (26 men and 4 women; mean age 61 years) who underwent PTCA, and another 49 patients (39 men and 10 women; mean age 63 years) who underwent diagnostic coronary angiography were enrolled in this study. Serum BNP concentrations were assayed in all patients. RESULTS: Plasma BNP was increased significantly with a peak concentration of 66.1 +/- 65.2 pg/ml 24 h after PTCA. Coronary angiography did not cause plasma BNP increase (immediately before 30.4 +/- 29.0 pg/ml, 24 h after 33.7 +/- 30.6 pg/ml). No significant differences were present in hemodynamic parameters measured immediately before and 24 h after PTCA. CONCLUSION: Plasma BNP is increased by transient myocardial ischemia induced by PTCA. PMID- 11061058 TI - Assessment of left ventricular systolic function in patients with idiopathic mitral valve prolapse using dobutamine stress echocardiography. AB - BACKGROUND: Some previous studies performed with radionuclide ventriculography and thallium scintigraphy reported that patients with idiopathic mitral valve prolapse (MVP) had some degree of left ventricular (LV) systolic dysfunction and that this dysfunction was more commonly found in symptomatic patients. HYPOTHESIS: The aim of the present prospective study was to investigate LV systolic function and its relationship with symptoms in patients with MVP with dobutamine stress test without associated certain mitral regurgitation and coronary artery disease. METHODS: Thirty-three patients with echocardiographically diagnosed idiopathic MVP were enrolled into the study and were divided into two groups as symptomatic (MVP-s) and asymptomatic (MVP-a). Patients underwent dobutamine stress echocardiography (DSE) to determine wall motion abnormalities and ejection fraction (EF) changes during rest state and increased heart rates. Results were compared with the DSE findings of 25 healthy individuals. RESULTS: Symptomatic patients (MVP-s) had lower EFs during the pretest period than the control group (59.0 +/- 4.8% and 68.3 +/- 5.7%, respectively, p < 0.05). Basal wall motion abnormalities were found in one patient in the MVP-a group (6%) and in two patients in the MVP-s group (12%). During DSE, new wall motion abnormalities (inferoapical dyskinesia) occurred in two patients in the MVP-s group at submaximal heart rates. For EF values calculated when patients reached submaximal heart rate, the MVP-s group showed only a 2.7 +/- 3.1% increase from baseline values. This increase was 5.1 +/- 3.8% in the MVP-a group and 9.3 +/- 4.3% in the control group (p < 0.05 between MVP-s and control groups). CONCLUSION: There is a close relationship between symptoms and ventricular function in patients with idiopathic MVP, and although many asymptomatic patients had nearly normal LV function, a subgroup of symptomatic patients showed diminished LV function and wall motion abnormalities. PMID- 11061059 TI - Atherosclerotic coronary artery aneurysm. PMID- 11061060 TI - Effects of theophylline on exercise indices in a patient with chronotropic incompetence. AB - Several investigators have documented the successful use of oral sustained release theophylline in treating symptomatic bradycardia and sick sinus syndrome. This paper reports a case of chronotropic incompetence in which specific exercise indices, including the chronotropic response index, were used to measure the therapeutic efficacy of theophylline. PMID- 11061061 TI - Transcatheter closure of a patent ductus arteriosus in an elderly patient with the Gianturco-Grifka vascular occlusion device. AB - This paper reports the unusual case of a 76-year-old woman who was discovered to have a hemodynamically significant patent ductus arteriosus following uneventful mitral valve replacement, utilizing cardiopulmonary bypass. The shunt was successfully eliminated using a new transcatheter device, thereby obviating the need for further surgery. The patient did not have calcification within the ductus, making diagnosis prior to surgery more difficult. PMID- 11061062 TI - Giovanni Battista Morgagni and the foundation of modern medicine. PMID- 11061063 TI - Diagnosis and treatment of common sexually transmitted diseases in women. AB - Chlamydia trachomatis and Neisseria gonorrhoeae are 2 very common sexually transmitted organisms, whose clinical manifestations in women can range from an asymptomatic carrier state to active pelvic inflammatory disease with known serious sequelae, including chronic pelvic pain, infertility, and ectopic pregnancy. The economic and clinical burden of these 2 infectious organisms are significant in the sexually active population. New developments in diagnosis and treatment of these infections raise great hope that substantial reduction in morbidity and disease prevalence rates can be achieved. Herpes simplex virus is probably better publicized and more feared in the sexually active population, and is far more prevalent than previously recognized; fortunately, however, it is not generally associated with significant morbidity. This article will review the current diagnoses and treatments of these conditions and consider some of the issues surrounding the impact of screening asymptomatic sexually active individuals. The treatment guidelines will emphasize the 1998 Guidelines for Treatment of Sexually Transmitted Diseases from the Centers for Disease Control and Prevention. PMID- 11061064 TI - Workup of the abnormal Pap test. AB - Cytologic evaluation of cells obtained from the cervix and vagina was first proposed by Papanicolaou and Traut in the 1940s as a method of detecting cervical cancer and its precursor lesions. Since that time, cervical cytology has proved to be the most efficacious and cost-effective method of cancer screening. By increasing detection of preinvasive and early invasive disease, use of Papanicolaou's (Pap) test has decreased both the incidence and mortality of cervical cancer in areas with well-established screening programs (1). The American Cancer Society has estimated that in the United States, cervical cancer will be diagnosed in 14,900 women and 4500 women will die as a result of this disease during the year 2000. More than 50 million Pap tests are performed annually in the United States, and about 5% of them will be abnormal (2). Consequently, it is incumbent on the practicing primary care physician to be familiar with the clinical significance and natural history of abnormal cervical cytologic diagnoses as well as the available treatment options. This discussion will delineate practical management protocols for the full range of cervical dysplasia commonly encountered in clinical practice. PMID- 11061065 TI - Abnormal genital tract bleeding. AB - The etiology of abnormal genital tract bleeding encompasses a wide range of disorders that can be secondary to anatomic changes of the female genital tract, infection, endocrinologic disorders, malignancies, and systemic illness. Appropriate workup is guided by age-related differential diagnoses for abnormal bleeding. Modern diagnostic tools can quickly focus the evaluation and allow timely intervention. Most abnormal genital tract bleeding is uterine bleeding, which is one of the most common gynecologic problems that health care providers will face. It accounts for approximately 15% of office visits and 25% of gynecologic operations. Abnormal uterine bleeding in reproductive-age women is defined as bleeding at abnormal or unexpected times or by excessive flow at the time of an expected menses. The average menstrual cycle length and duration of flow is 28 days and 4 days, respectively, with an average blood loss of 35 cc (1). Any bleeding should be considered abnormal in premenarchal girls and in post menopausal women except for those with predictable withdrawal bleeding taking hormone replacement therapy. This article will review the categories of abnormal genital tract bleeding and the diagnostic tools needed to establish the correct diagnosis. Common clinical cases will be presented to illustrate the presenting symptoms, differential diagnoses, workup, treatment, and long-term follow-up. PMID- 11061066 TI - Vaginitis: meeting the clinical challenge. AB - Vaginitis is a common gynecologic disorder that is responsible for 10 million office visits to physicians each year. Infectious vaginitis is the most common cause of a vaginal discharge, but other important diagnostic considerations include infectious cervicitis, a physiologic discharge, atrophic vaginitis, and allergic or irritant vaginitis. Although the history and gynecologic examination may suggest the diagnosis, laboratory confirmation should be routinely sought by performance of the vaginal pool wet mount examination, the amine whiff test, determination of the vaginal pH, and the Q-tip test. Once a precise diagnosis is made, effective therapy can then be prescribed. For patients with Candida vaginitis, therapeutic options include either the vaginal administration of a number of available imidazole or triazole antifungal agents or the prescription of the oral triazole agent fluconazole. Oral metronidazole remains the only effective treatment for trichomoniasis in the United States. Bacterial vaginosis, which has been linked to a number of obstetric and gynecologic complications, is effectively treated with oral metronidazole, although vaginal metronidazole gel and oral and vaginal clindamycin formulations are available as well. PMID- 11061067 TI - Contraceptive update Y2K: need for contraception and new contraceptive options. AB - Despite the major strides made in birth control, which have produced a decline in unintended pregnancies over the past decade and the lowest rates of teen pregnancies seen since 1974 (1,2), significant problems still remain. Almost half (48%) of US pregnancies in 1995 were unintended (1) and many more that were "intended" were not planned or prepared for (3). To optimize maternal and fetal outcomes, it is incumbent that physicians both emphasize the need for women to be physically, emotionally, and socially prepared for pregnancy before they conceive as well as ensure the availability of effective methods to allow them to do so. Today, contraceptives are available that permit couples to choose if and when to have children. Although only 5% of women who are sexually active and say they do not want to become pregnant are using no method of birth control (4), that group accounts for nearly 40% of the unintended pregnancies. More than half of all unintended pregnancies occur in women who had used a method in the month of conception (1). The strategy with these women should be to find ways to make the method they select work better for them or to switch them to more effective methods. Unfortunately, the most effective reversible methods are among the least utilized--in part because they have the highest initial costs. Some states, such as California and Maryland, have passed Contraceptive Equity Acts, which require insurance companies that provide any prescriptive drug coverage to cover all forms of prescription contraception. Many other states, as well as the federal government, are now considering similar legislation. It is important, therefore, both from the perspective of quality patient care and also from a fiscal standpoint, that all who care for reproductive-aged women become familiar with the full array of contraceptive options. This article will review the methods of reversible birth control now available in the United States, including the most recent efficacy rates and new practical tips to achieve better utilization, as well as summarize the features of some of the new options that may be nearing introduction. PMID- 11061068 TI - Incidence of exit-site infection with various exchange systems in continuous ambulatory peritoneal dialysis. AB - BACKGROUND AND PURPOSE: The disconnect twin-bag (TB) system was first introduced in Taiwan for use as an exchange system in continuous ambulatory peritoneal dialysis (CAPD) in 1995. Following its introduction, the incidence of CAPD associated peritonitis declined, but the incidence of exit-site infection (ESI) increased. To determine the cause of the increase in ESI incidence after the introduction of the TB system, this study compared the incidence of ESI among patients using the O set, ultraviolet antiseptic (UV) device, and the TB system. METHODS: A total of 170 patients who had received CAPD for more than 3 months were enrolled in this study. Poisson test and Kaplan-Meier survival analysis were used to compare the ESI incidence and ESI-free catheter survival among patients using the O set, UV device, or TB system. Cox stepwise forward proportional hazard analysis was used to assess the impact of sex, education, cause of uremia, age, and type of exchange system on ESI. RESULTS: The incidences of ESI differed significantly among patients using the three exchange systems, with 20.9, 13.8, and 4.0 episodes per 100 patient-years for patients using the TB system, O set, and UV device, respectively. New patients using the TB system also had a shorter mean interval of ESI-free catheter survival than those using the UV device (26.9 vs 58.8 months, p = 0.002). In the Cox stepwise forward proportional hazard analysis, non-lupus patients had a lower risk of developing ESI than lupus patients (relative risk [RR] 0.40, p = 0.03). The RR of ESI in patients using the UV device was also lower than in those using the TB system (RR 0.15, p < 0.01). CONCLUSION: In this study, use of the TB system was associated with a higher incidence of ESI. The increased ESI incidence may be related to the heavier mini transfer set of the TB system. Therefore, special attention should be given to fastening the mini-transfer set tightly during the exchanging procedure to prevent traction on the exit-site, which is associated with an increased incidence of subsequent ESI. PMID- 11061069 TI - Prevalence of urinary incontinence and intention to seek treatment in the elderly. AB - BACKGROUND AND PURPOSE: Urinary incontinence (UI) is a common, distressing, and often disabling condition in the elderly. The objectives of this study were to estimate the prevalence and clinical characteristics of UI among elderly individuals living at home and to explore their perceptions of UI and intention to seek medical care. METHODS: A total of 504 elderly subjects aged 65 and older residing in Tungkang town (located in the southwestern part of Taiwan) were randomly sampled and surveyed face to face by registered nurses. The prevalence, clinical types, and perceptions of UI, and intention to seek treatment, were compared with chi-square statistics across various sociodemographic characteristics. Logistic regression analyses were conducted to identify factors associated with UI experience and intention to seek treatment. RESULTS: About 22% of respondents reported that they had experienced involuntary loss of urine in daily life. Women, people who were overweight, and those who were aged 70 years or older were at higher risk of UI. While women were more likely to suffer from stress incontinence, men were at higher risk of urge incontinence. Women, illiterate individuals, and those who perceived UI as a normal part of the aging process showed low intention to seek treatment for UI. CONCLUSIONS: The results of this study suggest that public awareness programs about UI and promotion of available treatment options are necessary to increase the intention to seek treatment among the elderly. Culturally sensitive programs should be designed, particularly for female and illiterate elderly, to provide incentives to seek medical care. The increasing availability of various treatment modalities coupled with education to correct commonly held misconceptions about UI might enable more elderly individuals to receive treatment for this common condition. PMID- 11061070 TI - Locoregional recurrence in patients with one to three positive axillary nodes after mastectomy without adjuvant radiotherapy. AB - PURPOSE: To retrospectively assess the risk of locoregional recurrence (LRR) and analyze the prognostic factors of this pattern of failure in patients with breast cancer and one to three positive axillary lymph nodes treated with modified radical mastectomy (MRM) without adjuvant radiotherapy. METHODS: From April 1991 through December 1997, 649 patients received a diagnosis of invasive breast cancer, and 545 were treated with MRM. Eighty-one of these patients who were found to have one to three positive axillary nodes and had a minimum follow-up of 2 years were included in this study. None of the 81 patients received adjuvant radiation therapy after mastectomy; 43 patients received adjuvant chemotherapy; and 60 patients received adjuvant hormone therapy. The median duration of follow up was 39 months. RESULTS: Thirteen patients had LRR during follow-up, all within 2 years after mastectomy. The 3-year LRR rate was 14%. The 3-year rates of distant metastasis for patients with and without LRR were 48% and 14% (p = 0.03), respectively. The 3-year survival rates for patients with and without LRR were 73% and 87% (p = 0.01), respectively. In univariate analysis, age (p = 0.01), estrogen receptor (ER) status (p = 0.02), and the addition of hormone therapy (p < 0.001) were significant risk factors for LRR; in multivariate analysis, negative ER status (p = 0.02) was the only statistically significant risk factor. The 3-year LRR rates for ER-negative patients and those with positive or unknown ER status were 31% and 11%, respectively. CONCLUSIONS: LRR after mastectomy is a substantial clinical problem, despite the use of adjuvant chemotherapy and/or hormone therapy. Further randomized trials of postmastectomy radiotherapy for patients with one to three positive axillary nodes and specific risk factors are urgently needed to determine its potential benefit in locoregional control and survival, especially for young and ER-negative patients. PMID- 11061071 TI - Ectopic atrial tachycardia in children. AB - BACKGROUND AND PURPOSE: Ectopic atrial tachycardia (EAT) is an unusual and potentially risky arrhythmia that can result in left ventricular dysfunction if not properly managed. In adults, EAT is mainly caused by diseased atrial myocardium and responds poorly to antiarrhythmic drugs. The characteristics of EAT in children might be different from those in adults because of their immature myocardium and the different electrophysiologic characteristics of their conduction tissue. We examined the natural history and treatment of EAT in children. METHODS AND RESULTS: From June 1990 through June 1999, 24 children (8 girls and 16 boys; median age 4 mo [1 d-10 yr]) were admitted to our hospital with a diagnosis of EAT. Fifteen had healthy hearts, six had congenital heart disease, one had myocarditis, one had bronchopulmonary dysplasia with severe pulmonary hypertension, and one had hypertrophic cardiomyopathy. Thirteen patients presented with congestive heart failure. Only four patients had symptoms of prodromal airway infection. The maximum atrial rate was 244 +/- 66 beats per minute. Atrioventricular block was documented at least once during tachycardia in 10 patients. Warm-up or cool-down phenomenon was seen at the initiation or termination of tachycardia in thirteen patients. Primary pharmacologic treatment was attempted in all patients. EAT was initially controlled in all patients using digoxin plus propranolol (18 patients), propranolol only (4), or digoxin plus procainamide (2). EAT was controlled using medication in 22 patients. Fifteen patients had sinus rhythm but did not receive medication for 39 +/- 25 months. Two patients died of uncontrolled arrhythmia, and two of underlying disease. Recurrence was seen within 3 months after initial therapy in five patients. Surgery was performed to correct the underlying heart disease in three patients with frequently recurring EAT, all of whom remained tachycardia-free after surgery, without pharmacologic treatment. The spontaneous remission rate was 75% (18/24). CONCLUSION: EAT in children without underlying heart disease can be effectively treated using antiarrhythmic drugs. Spontaneous resolution of EAT after medication in children was frequent (75%) in this series. The results of this study suggest that a step-wise approach using digoxin, a beta-blocker, and a class I antiarrhythmic drug may be the most effective treatment for EAT. PMID- 11061072 TI - Urinary uric acid/creatinine ratio as an additional marker of perinatal asphyxia. AB - PURPOSE: To study the validity of urinary uric acid (UA) as a marker of perinatal asphyxia in term and premature infants. METHODS: The urinary ratio of UA to creatinine (Cr) was obtained within 24 hours after birth in four groups of infants: 17 term infants and 18 premature infants with perinatal asphyxia, and 22 healthy term infants and 20 premature infants without perinatal asphyxia. Perinatal asphyxia was defined as an Apgar score of 3 or less at 1 minute or 5 or less at 5 minutes, and/or a first blood gas pH of less than 7.25 and a base deficit of at least 12 mmol/L. RESULTS: The urinary ratio of UA to Cr was significantly higher in term infants with perinatal asphyxia than in term infants without asphyxia (1.53 +/- 0.71 vs 0.73 +/- 0.45; p < 0.005). The same result was found between premature infants with and without perinatal asphyxia (3.89 +/- 1.84 vs 2.45 +/- 0.88; p < 0.01). The urinary ratio of UA to Cr in premature infants was significantly higher than in term infants. When the urinary ratio of UA to Cr was greater than 0.95, perinatal asphyxia was identified with a sensitivity of 80% and a specificity of 71% in term infants. In premature infants, a cut-off value of UA/Cr for perinatal asphyxia of 2.9 had a sensitivity of 71% and a specificity of 70%. CONCLUSIONS: The results of this study indicate that the urinary ratio of UA to Cr may be used as an additional marker of perinatal asphyxia in term and premature infants. In comparison with other markers such as xanthine, hypoxanthine, and ascorbic acid, it is a simple, quick, and inexpensive way to detect hypoxic episodes in a neonatal intensive care unit within 24 hours after birth. PMID- 11061073 TI - Meningeal involvement in B-cell chronic lymphocytic leukemia: report of two cases. AB - Symptomatic central nervous system (CNS) involvement in chronic lymphocytic leukemia (CLL) or its variants is rare. We report two cases of CLL with leptomeningeal involvement. Patient one was an 81-year-old male who had CLL stage C (IV) at diagnosis and developed meningeal disease 29 months later. Patient 2 was a 42-year-old male with a diagnosis of CLL stage A (II) that evolved into mixed-cell CLL/prolymphocytic leukemia (PLL) 1.5 years later, with leptomeningeal infiltration of prolymphocytes developing 26 months after initial diagnosis. Meningeal leukemia was diagnosed by cerebrospinal fluid examination, with flow cytometry showing the same immunophenotypic findings of lambda-light chain restriction as the lymphocytes in bone marrow in one patient, and with morphologic characteristics exhibiting exclusively prolymphocytes in the other patient. The CNS disease of both patients responded effectively to intrathecal chemotherapy and cranial irradiation. However, both patients died of infection, a major cause of morbidity and mortality in patients with CLL. The clinicopathologic features of these two patients indicate that, despite the rarity of CNS involvement in CLL patients, any neurologic manifestation in CLL patients should arouse suspicion of meningeal leukemia and patients should be examined and managed accordingly. PMID- 11061074 TI - Tuboovarian abscess mimicking malignancy: report of two cases. AB - Tuboovarian abscess is a well-known sequela of acute or chronic salpingitis. In a small percentage of patients, these inflammatory masses compress or even rupture into the adjacent viscera, thus simulating the condition of pelvic malignancy, particularly when the clinical presentations are indolent. We describe two cases of tuboovarian abscess mimicking malignancy. Case 1: A 39-year-old woman with an intrauterine device had a clinical presentation mimicking an exophytic submucosal colorectal tumor with suspicious mucosal invasion. She complained of tenesmus but did not experience fever or adnexal tenderness. A right tuboovarian abscess with fistula formation into the rectosigmoid colon was noted during laparotomy. Case 2: A 46-year-old woman with an intrauterine device had a preoperative diagnosis of uterine myoma with degeneration. At laparotomy, an omentum cake with dense pelvic adhesions was noted. Malignancy appeared to be present, and debulking surgery was performed. The final pathologic examination revealed bilateral chronic tuboovarian abscesses and focal omental abscess. PMID- 11061075 TI - Infected chylothorax caused by Streptococcus agalactiae: a case report. AB - Chylothorax is bacteriostatic in nature. Bacterial infection rarely develops in chylothorax and has never been reported in a non-immunocompromised host. A 33 year-old woman was admitted to National Taiwan University Hospital because of fever and right pleuritic pain. Chest roentgenography and computed tomography revealed right pleural effusion. Examination of the pleural effusion revealed a profile compatible with empyema and chylothorax. Culture of the pleural effusion yielded Streptococcus agalactiae. The woman was not immunocompromised. This is the first report of infected chylothorax caused by Streptococcus agalactiae in a non-immunocompromised host. PMID- 11061076 TI - Sick sinus syndrome in a patient with single coronary artery anomaly. AB - Single coronary artery anomaly is very rare. The reported manifestations include angina pectoris and congestive heart failure. Here we describe a case of single coronary artery anomaly presenting as sick sinus syndrome, which has no literature precedence. A 47-year-old woman had complained of intermittent dizziness for years. A Holter electrocardiogram showed sinus bradycardia and junctional or ventricular rhythm with a maximal ventricular pause of up to 3.2 seconds. Electrophysiologic study revealed prolonged corrected sinus nodal recovery time. Coronary angiography showed that the left anterior descending artery had a long course with a side branch originating from the proximal part and coursing anteriorly to the territory of the proximal portion of the right coronary artery. The sinus node is usually supplied by the sinoatrial branch via the right coronary artery. Aortography showed that the right coronary artery ostium was absent. A permanent pacemaker was implanted and the patient was discharged in good condition. The present case suggests that coronary artery anomaly may lead to compromised blood supply to the sinus node, and hence sick sinus syndrome. PMID- 11061077 TI - gamma-sarcoglycan deficiency muscular dystrophy in two adults. AB - All dystrophin-associated proteins contain sarcoglycan complex. Different forms of muscular dystrophy are caused by defective expression of different proteins of this structure. gamma-Sarcoglycan deficiency muscular dystrophy, so-called severe childhood autosomal recessive muscular dystrophy (SCARMD), is a rare disease that has not been previously reported in Taiwan. This paper describes two Taiwanese adults with this disease: a 26-year-old man with calf pseudohypertrophy who had weakness in both legs for 1 year; and a 43-year-old woman who had progressive weakness in all four limbs, with the initial symptom of gait disturbance at the age of 32 years. Analysis of muscle biopsy specimens, which showed total deficiency of gamma-sarcoglycan protein on immunostaining, confirmed the diagnosis of SCARMD in both cases. However, the clinical manifestations in these two patients, including lower proximal limb weakness initially developing in adulthood with a slow progressive course, are different from previously reported cases of SCARMD. The literature on this disease is reviewed and possible mechanisms of these distinct clinical presentations are discussed. PMID- 11061078 TI - Acute lymphoblastic leukemia in one of two siblings with Alstrom syndrome. AB - Alstrom syndrome is a rare autosomal recessive disease; less than 60 cases have been reported. No Chinese patient with this disease has been reported previously in the literature. Here, we describe an 11-year-old Chinese boy with this condition. His elder sister also had Alstrom syndrome, and his father had non insulin-dependent diabetes mellitus. Both siblings had degenerative retinopathy, obesity, mental retardation, perceptive hearing loss, short stature, non-insulin dependent diabetes mellitus, nephropathy, hyperlipidemia, acanthosis nigricans, and hepatic dysfunction. The boy also developed acute lymphoblastic leukemia, which was confirmed by cytochemistry and immunophenotyping findings. He received chemotherapy and radiotherapy for the malignancy. The present case suggests that acute lymphoblastic leukemia may be coincident with or may be a previously undescribed systemic manifestation of Alstrom syndrome. PMID- 11061079 TI - Renal malformations in children with Turner's syndrome. AB - Between 1988 and 1999, renal sonography and intravenous urography were performed to detect renal malformations in 54 patients with Turner's syndrome (TS). The mean age of these patients at diagnosis of TS was 9.2 +/- 4.6 years. Renal malformations were detected in 21 patients by intravenous urography and there was no significant difference in the frequency of renal malformations among different karyotype groups. Horseshoe kidney was the most common renal malformation, followed by duplex kidney. Fifteen of 21 renal malformations were not detected by renal sonography. We conclude that these TS patients had a high frequency of renal malformations, and that the detection rate of horseshoe kidney and duplex kidney by renal sonography was not satisfactory. Although renal sonography alone can be used to detect more severe renal malformations that may need further management, it may underestimate the frequency of renal malformation in children with TS. PMID- 11061080 TI - [The value of serum C-reactive protein as a survival determinant in patients with advanced non-small-cell lung cancer]. AB - We retrospectively reviewed the information in our data base concerning 127 patients with advanced non-small-cell lung cancer. Seventeen pretreatment clinical variables including serum C-reactive protein (CRP) were analyzed to determine the factors related to survival using Cox's proportional hazards model. Univariate analysis revealed that eleven explanatory variables were significant. In multivariate statistical technique for those variables, six were selected as significant factors. The highest hazard ratio was observed in the serum CRP (3.82). The other factors were therapy (2.52), serum lactate dehydrogenase (2.41), serum total protein (2.20), white blood cell counts (1.98) and performance status (1.80). Median survival times estimated by the Kaplan-Meier procedures in patients with normal CRP (CRP < 0.2 mg/dl) and high positive CRP (CRP > or = 3.0 mg/dl) were 24.9 months and 3.7 months, respectively. These results suggest that serum CRP is an independent survival determinant in advanced non-small-cell lung cancer. PMID- 11061081 TI - [Impact of discontinuing peak flow monitoring in stable asthma]. AB - We conducted a prospective study to examine the influence of discontinuing peak flow monitoring (PFM) in stable asthmatics who had already been properly educated and were monitoring their own peak expiratory flows (PEF). All subjects had been performing PFM for at least 3 months prior to their entry into the study, and PFM was then stopped for a period of 3 months. Comparisons of endpoints were made between a period of 3 months prior to, and after discontinuing PFM. Forty patients with a mean age of 52 were studied. Only one patient experienced a single emergency room visit either before or after discontinuing PFM. Short courses of oral steroids were administered in 6 patients (15%), both before and after discontinuing PFM. There was no significant change in pulmonary function, beta 2-agonist use and asthma symptoms during a 3-month period before and after discontinuing PFM. These results suggest that simply continuing PFM in stable, educated asthmatics may have little impact on asthma control, although its long term influence should be carefully examined. PMID- 11061082 TI - [Role of nitric oxide in airway mucociliary dysfunction in diffuse panbronchiolitis]. AB - Nitric oxide (NO) stimulates the ciliary motility of the airway epithelium, thereby assisting in the regulation of mucociliary transport in the respiratory tract. In the present study, to elucidate a possible involvement of NO in mucociliary disturbance in diffuse panbronchiolitis (DPB), we measured nasal clearance time (NCT) using the saccharin test, NO concentration in the exhaled air with a chemiluminescence analyzer, and electrolyte concentration in the sputum. Compared with healthy nonsmokers and smokers, patients with DPB showed a lower NO concentration in the exhaled air (p < 0.05), prolonged NCT (p < 0.01), and a higher Cl concentration in the sputum (p < 0.05). Among these variables, exhaled NO concentration was negatively correlated with NCT (p < 0.01) and sputum Cl concentration (p < 0.05). These results suggest that the inhibition of NO generation associated with an elevated Cl concentration in the airway surface liquid may contribute to airway mucociliary dysfunction in DPB. PMID- 11061083 TI - [A case of idiopathic pulmonary hemosiderosis of adult onset]. AB - A 51-year-old man presenting with hemoptysis was admitted to our hospital. Chest radiography revealed air space consolidation in the right lung field. Laboratory data showed anemia, hypoxemia, and no evidence of inflammatory signs, bleeding tendency, renal dysfunction, or collagen vascular diseases. Tests of anti-GBM antibody, P-ANCA, and C-ANCA were negative. Microscopic examination of the lung tissue specimens obtained by video assisted thoracic surgery revealed hemorrhage and numerous hemosiderin-laden macrophages in the alveoli. No deposition of immunoglobulin and vasculitis were seen. These findings were consistent with a diagnosis of idiopathic pulmonary hemosiderosis. Steroid therapy had a limited effect, and the patient died. Idiopathic pulmonary hemosiderosis of adult onset is rare in Japan. PMID- 11061084 TI - [A case of multidrug-resistant pulmonary tuberculosis]. AB - A 44-year-old woman with multidrug-resistant pulmonary tuberculosis was admitted to our hospital in August 1998. She had been treated with the anti-tuberculosis agents isoniazid (INH), rifampicin (RFP), pyrazinamide and streptomycin (SM) for two months. However, tubercule bacilli found in a sputum culture on admission showed resistance to INH, RFP and SM, and so these agents were replaced with kanamycin (KM), ethionamide, cycloserine and levofloxacin. Unfortunately, the bacilli persisted in the sputum smears, and the patient complained of prolonged pain in the sites of intramuscular injection of KM. In January 1999, inhalation of KM was begun, resulting in the disappearance of the bacilli from the sputum and in improvements in chest radiographs. Inhalation of KM could be an effective therapy, with fewer adverse effects, in cases of multidrug-resistant pulmonary tuberculosis. PMID- 11061085 TI - [Hypersensitivity pneumonitis caused by Lyophyllum aggregatum in two sisters]. AB - We encountered two sisters with hypersensitivity pneumonitis caused by Lyophyllum aggregatum. In Case 1, the patient was a 47-year-old woman who consulted Iiyama Red Cross Hospital because of a non-productive cough. She had worked with Lyophyllum aggregatum cultures for 7 years. In 1996, she complained of a dry cough, and had had a severe cough in November 1998. She was admitted to Iiyama Red Cross Hospital because of dyspnea on effort. Laboratory data revealed a high level of C-reactive protein. Chest radiography and CT scanning showed ground glass opacities in both lower lobes, and centrilobular nodules. Bronchoalveolar lavage fluid revealed a decrease in the CD4/CD8 ratio, and the lymphocyte fraction was very high. Transbronchial lung biopsy revealed lymphocytic granuloma and interstitial thickening, and the lymphocyte stimulation test was positive for Lyophyllum aggregatum. Case 2 occurred in the 51-year-old sister of Patient 1. She had worked with Lyophyllum aggregatum cultures for 10 years. In 1994, she had experienced dyspnea and a dry cough while working. In January 1999, she was admitted to Iiyama Red Cross Hospital because of a dry cough, sputum and chest oppression. Chest radiography and CT scanning revealed ground glass opacities in both lower lobes and centrilobular nodules. The CD4/CD8 ratio in the bronchoalveolar lavage fluid was low, and the lymphocyte fraction conspicuously high. The transbronchial lung biopsy revealed lymphocytic granuloma and interstitial thickening. The lymphocyte stimulation test was negative for Lyophyllum aggregatum. She was discharged and returned to work, but the dry cough and low-grade fever recurred. However, the occupational provocation test is not regarded as positive because chest radiography, laboratory examinations, and a pulmonary function test were not performed. The symptoms of both cases were eliminated only on isolation of the antigen. There has not been any previous report of hypersensitivity pneumonitis caused by Lyophyllum aggregatum. PMID- 11061086 TI - [Clinical study of three myelodysplastic syndrome (MDS) patients with BOOP-like pulmonary disease]. AB - Three cases of myelodysplastic syndrome (MDS) associated with BOOP-like pulmonary disease were reported. They were diagnosed from transbronchial biopsies and clinical features. In two cases, chest radiographs showed ground glass opacities or air space consolidation in both lung fields, and air space consolidation in the right lower lobe in the other case. BALF showed a marked increase of lymphocytes in one case, and organizing pneumonia and alveolitis, and alveolar spaces filled with foamy macrophages were identified histologically. Although no steroid therapy was employed, all three cases improved. However, one patient suffered a relapse 6 months later and thereafter did not respond to corticosteroid therapy. PMID- 11061087 TI - [A case of radiation lung injury with characteristics of bronchiolitis obliterans organizing pneumonia after radiation therapy for breast carcinoma]. AB - We report a case of sputum and dyspnea with patchy migratory air space infiltrates that developed in a 49-year old woman after she started breast radiation therapy following surgery for breast carcinoma. Our case clearly differed from ordinary radiation pneumonitis. Chest roentogenography and computed tomographic (CT) scanning demonstrated alveolar opacities. Bronchoalveolar lavage showed a moderate elevation of the total cell concentration and a considerable increase of lymphocytes, and transbronchial lung biopsies revealed a histological pattern of bronchiolitis obliterans organizing pneumonia with intra-alveolar granulation tissue. The present case suggests that breast irradiation may contribute to the development of a histological pattern of bronchiolitis obliterans organizing pneumonia. PMID- 11061088 TI - [Three cases of measles-associated interstitial pneumonia]. AB - It has been reported that measles-associated pneumonia is easily complicated with fatal respiratory failure. During the 6 months from January to June 1998, we treated three young patients with measles-associated pneumonia who had previously been healthy. Their chest X-ray and CT examinations revealed mainly interstitial and partially alveolar pneumonia surrounding extensive areas of bronchiolitis. In one patient, respiratory failure was present, and arterial blood gas analysis showed severe hypoxemia. Therefore, this patient was treated with a mechanical ventilator, which dramatically improved the condition. The other two patients received steroid-pulse therapy and improved rapidly. We believed that in measles associated pneumonia, steroid pulse therapy is to be recommended for the severely ill patient. According to a report by the Japanese Ministry of Health and Welfare, measles-like disease was widespread in Japan in 1991 and occasionally encountered in 1996. But in Yamanashi Prefecture, the disease was prevalent in 1991, 1994 and 1998. The number of patients in our hospital had reflected the prevalence in the prefecture. It is suggested that the measles is a disease of regional prevalence. PMID- 11061089 TI - [A case of primary pulmonary artery myxosarcoma associated with severe pulmonary hypertension]. AB - A 50-year-old man presented with progressive dyspnea on exertion, but with no history of chest pain or syncope. Chronic pulmonary thromboembolism was suspected and he was referred to our hospital. On ausculation, a grade 3 systolic murmur was heard, that was loudest in the fifth intercostal space lateral to the right sternal border. Chest radiography showed mild cardiomegaly and ventilation perfusion scan revealed absence of perfusion in the left lung and the upper field of the right lung. Contrast-enhanced helical CT showed large mural defects in both main pulmonary arteries, clearly delineated by contrast medium. The left pulmonary artery was nearly completely occluded, and eccentric defects were observed projecting into the lumen of the pulmonary trunk. A tumor originating in the pulmonary artery was suspected, but a definitive diagnosis of the mass could not be made with pulmonary angiography and magnetic resonance imaging. The mean pulmonary arterial pressure was 50 mmHg. Further radiologic examinations failed to reveal the source of the embolus or tumor. It was decided to attempt surgical excision under total cardiopulmonary bypass. At operation, a gelatinous, lustrous, yellowish mass was found partially occluding the right main pulmonary artery and completely occluding the left. The tumor adhered tightly to the intima of the vessel and was inoperable. The patient could not be weaned from percutaneous cardiopulmonary support and died 3 days after surgery. Histologic examination of the excised specimen revealed myxosarcoma. PMID- 11061090 TI - [A case of Sjogren's syndrome with pleural effusion]. AB - A 45-year-old woman was admitted to our hospital because of a fever. A round erythema was noted on the skin, suggesting collagen disease. Bilateral pleural effusion developed during hospitalization, and serum and pleural effusion were positive for antinuclear antibody, RA factor, anti-SS-A antibody, and anti-SS-B antibody. A diagnosis of Sjogren's syndrome was made on the basis of reduced lacrimation and the histological findings in a biopsy specimen from the lip. The cells in the pleural effusion were predominantly lymphocytes, and so a pleural lesion associated with Sjogren's syndrome was suspected, but reports of this condition have been scarce. Good therapeutic results were obtained by corticosteroid administration. Sjogren's syndrome should be considered in the differential diagnosis of pleural effusion associated with collagen disease. PMID- 11061091 TI - [Pneumocystis carinii pneumonia associated with acquired immunodeficiency syndrome followed by KL-6, surfactant protein-D and beta-D-glucan in serum]. AB - A 40-year-old man was admitted to our hospital with acute respiratory failure. The patient was given a diagnosis of Pneumocystis carinii pneumonia (PCP) associated with acquired immunodeficiency syndrome (AIDS). After treatment with trimethoprim-sulfamethoxazole and corticosteroid, the respiratory failure was improved and the abnormal shadows disappeared. The serum beta-D-glucan level, significantly elevated (76.0 pg/ml) on admission, returned to the normal range within two weeks. Serum KL-6 (max. 7580 U/ml) and surfactant protein-D (SP-D) (max. 235 ng/ml), which are produced by type II pneumocytes, increased after elevation of the beta-D-glucan level and decreased gradually following successful treatment. These findings suggest that beta-D-glucan may be a serological marker for PCP infection and KL-6 may be a serological marker for lung injury in PCP with AIDS. PMID- 11061092 TI - [Respiratory failure due to diaphragmatic dysfunction in Charcot-Marie-Tooth disease: a case report]. AB - A 56-year-woman with type 2 respiratory failure due to diaphragmatic dysfunction in Charcot-Marie-Tooth disease (CMT) is reported. The patient, who had a 50-year history of CMT, was referred to our hospital because of nocturnal dyspnea. Arterial blood gas analysis on admission showed marked hypoxia with hypercapnia, and physical examination revealed thoracoabdominal paradoxus in the supine position. Chest radiography revealed elevation of both sides of the diaphragm. The vital capacity and arterial blood gas pressure in the sitting position were markedly higher than those in the supine position. Electrical phrenic nerve stimulation failed to produce any convincing muscle action potential in the diaphragm. These findings suggested that her respiratory failure was induced by both diaphragmatic dysfunction caused by bilateral phrenic nerve palsy due to CMT. Treatment of this patient was started at home with a pressure support ventilator, resulting in satisfactory clinical improvement. In general, respiratory muscle impairment is a rare phenomenon in a patient with CMT. However when a patient with CMT complains of dyspnea or if unexpected heart failure develops, it is important to keep in mind that CMT may be associated with phrenic nerve palsy. PMID- 11061093 TI - [Compensatory growth of residual lung after pneumonectomy in childhood]. AB - Though pneumonectomy proved to be a potent stimulus to compensatory growth in the animal model, the literature on this subject in human patients is sparse. We report a rare case of compensatory growth after pneumonectomy in childhood. A 17 year-old man was admitted to our hospital because of chest pain. He had been capable of normal exercise despite a history of pneumonectomy at the age of 4. Chest radiography showed pneumothorax and a displaced heart. In a chest CT scan, both pleural spaces were filled with the enlarged left lung. Since thoracoscopic examination showed a bulla at the surface of S3, bullectomy was performed. The specimen resected from the lung showed slight dilatation of the alveoli. It is considered that the enlargement of the residual lung had occurred as a result of alveolar multiplication rather than by dilatation of the existing alveoli. This case demonstrates lung regeneration in childhood. PMID- 11061094 TI - [The second chromosome in the account]. PMID- 11061095 TI - [Usefulness of fine needle aspiration biopsy in thyroid diseases]. PMID- 11061096 TI - [Lupus nephropathy at the beginning of the year 2000]. PMID- 11061097 TI - [Tropical sprue: a non-existent or overlooked disease?]. PMID- 11061098 TI - [Gene therapy for portal hypertension]. PMID- 11061099 TI - [Comparison of the usefulness of intraoperative examination and fine needle aspiration biopsy in thyroid lesions. Analysis of discordant cases in aspiration biopsy]. AB - INTRODUCTION: Fine-needle aspiration biopsy (FNAB) is currently considered the most reliable and cost effective examination for diagnosis of solitary thyroid nodules prior to surgery. Because of its great utility the indication of intraoperative examination (IOE) (macroscopic examination, cytology and frozen section), has recently been questioned. OBJECTIVE: To compare the accuracy of the FNAB and IOE, in those patients with nodular thyroid disease who undergo thyroidectomy. As well as to analyse the, discrepant cases by FNAB. MATERIAL AND METHODS: The results of IOE and FNAB were compared in a period of two years (1997 98) at the Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran (INCMNSZ). In all cases the diagnosis established from paraffin embedded tissue sections was considered to be the gold standard. The cytologyc smears and the histologic sections of the discrepant cases were reviewed. RESULTS: One thousand and fourteen IOE were done during two consecutive years; from these cases, 136 (13.4%) were thyroid lesions. Half of the thyroid lesions (69 cases) corresponded to carcinomas, the others were follicular adenomas (13 cases) and non-neoplasic lesions (54 cases). The analysis of the copacity to discriminate between malignant and benign diseases with IOE and FNAB demonstrated sensitivity of 89% (CI: 78.2-95.1) and 97.7% (CI: 86.8-99.9), specificity of 100% (CI: 93.1-100) and 90% (CI: 90.4-96.7). The positive predictive value of 100% and 91.6%, negative predictive value of 90.4% and 97.3%, and equal accuracy (94.6% and 94.1% respectively). When the analysis of capacity to stratify diseases was performed, IOE and FNAB showed similar percentages in all the cases, including the follicular adenomas and carcinomas. The causes of false positive and false negative cases by FNAB, reproduce what has been written in the literature. CONCLUSIONS: FNAB preoperative examination is adequate for the selection of patients that undergo thyroidectomy. Nevertheless, IOE should be done in most of the cases, and routinely in those cases with inconclusive FNAB results and in cases of follicular tumor. PMID- 11061100 TI - [Histopathology of sun prurigo]. AB - BACKGROUND: Solar or actinic prurigo (PS) is one of the most common skin diseases observed in our country, particularly in the pediatric age; in our institution it is on the 14 place with a frequency of 1.34%. It represents an abnormal reaction to solar light and in its pathogenesis diverse factors participate, mainly immunogenetical and environmental. Although their clinical characteristics are already specified, the histopathological features are still not well documented. MATERIAL AND METHODS: We carried out a retrospective and observational study in order to analyze the histopathologic changes in 20 samples of skin coming from 20 pediatric patients with PS; in all of them diagnosis was well established on clinical criteria. RESULTS: Twenty children with PS of more than one year of evolution, ten of each sex, all mestizos or indigenous inhabitants of Mexico City and vicinity; we identified recent and late lesions, with epidermal and dermal changes, the more conspicuous were: spongiotic papules usually with psoriasiform epidermal hyperplasia, in association with perivascular, superficial and mild lymphocytic infiltrates, usually with few eosinophils, exocytosis of lymphocytes into the epidermis, pigment incontinence and extravasation of erythrocytes. CONCLUSIONS: The PS is an inflammatory cutaneous disease for which histopathological diagnosis is feasible of being established on skin biopsies of recent or late lesions. PMID- 11061101 TI - [Clinical course and prognostic factors in lupus nephropathy]. AB - BACKGROUND: The present study was designed to know the clinical course of lupus nephritis and the risk factors associated with the development of end stage renal disease. METHODS: This a retrospective study performed in a cohort of 154 patients with biopsy proven lupus nephritis that were seen in our hospital between 1984 and 1990. The clinical records of all patients were reviewed in order to collect the following information at the time of the biopsy: age, sex, number and type of lupus criteria according with the American College of Reumathology, mean arterial pressure, serum creatinine, BUN, and albumin, as well as urinary protein excretion. The follow up was registered from the day the biopsy was performed to one of the following end points: end stage renal disease (defined as requirement of chronic dialysis), death or the end of study. All biopsies were analyzed by light microscopy to obtain the hystological subtype of lupus nephritis (WHO classification) and when type IV was diagnosed, the activity and chronicity indexes were also assessed. Kaplan-Meier survival tables were constructed. The association of clinical and laboratory variables with the development of end stage renal disease was obtained by log rank analysis. Variables obtained as significant were used to evaluate their individual impact using either the Cox multivariate proportional hazard method. RESULTS: Follow up was complete in 144 patients with a follow up time of 68 +/- 38 months. Ninety three patients were female with mean age of 28 +/- 9 years. At the time of the biopsy, renal manifestations had been present for 35 +/- 38 months and the number of lupus criteria per patient were 4 +/- 1. The clinical picture at the time of the biopsy was: nephrotic syndrome in 60%, non nephrotic proteinuria in 40%, and nephritic syndrome in only 2%. The hystological type of lupus nephritis was: I in 2%, II in 8%, III in 6%, IV in 71% and V in 11%. At the end of the study 28 patients developed end stage renal disease. For the whole group the survival of renal function was 85% at 70 months and 70% at 140 months. All, but one patient that developed end stage renal disease exhibited type IV nephropathy. In this subpopulation the mean activity and chronicity indexes were 8.5 +/- 3.5 and 3.1 +/- 2.4, respectively. By multivariate analysis the strongest predictors of end stage renal disease were the serum creatinine at the time of the biopsy, chronicity index, and age. The higher the serum creatinine and chronicity index at the time of biopsy, the higher the probability of developing end stage renal disease. CONCLUSIONS: We conclude that the clinical course of lupus nephritis in our population is similar to that seen in other series. The variables indicating advanced renal disease, such as high serum creatinine and chronicity index, were the strongest predictors of end stage renal disease. PMID- 11061102 TI - Validation of a prognostic index in the critically ill newborn. AB - OBJECTIVE: Elaborate and assess the degree of validity of a prognostic model for evaluating patients admitted to the Neonatal Intensive Care Unit (NICU). DESIGN: Cases and controls nested in a cohort. SETTING: NICU of two tertiary hospitals and another second level hospital. PATIENTS: The study was carried out in two phases. During the first phase (elaboration of a model), 336 newborns were studied; 112 belonged to the case group (dead patients) and 224 to the control group (live patients discharged). For the second phase (model validation), 300 patients were included that did not participate in the first phase, 100 cases and 200 controls. MEASUREMENTS: For each patient admitted to the study, clinical, paraclinical, perinatal and comorbidity factors were determined within the first 12 hours. Variables of statistical significance in the bivariate analysis were included in a logistic regression model with the objective of identifying a prognostic model. RESULTS: The variables that constituted the prognostic index were gestational age x birth weight, the paO2/FiO2 ratio x O2 saturation, arrest cardiac, major congenital malformations, septicemia and base excess. The model showed to have a sensitivity of 70% and a specificity of 91% during the elaboration cohort. In the validation cohort, sensitivity was 68% and specificity was 92%, a positive predictive value of 80%, negative predictive value of 85% and a correct classification rate was 84%. CONCLUSIONS: The Neonatal Mortality Prognostic Index (NMPI) developed in this study showed to be useful for the evaluation of hospital mortality for severely ill newborns admitted to NICU. PMID- 11061103 TI - Horizontal appendicular dysmetria in four patients with a typical Wallenberg's syndrome. AB - During the acute stage of a Wallenberg's syndrome ipsilateral appendicular dysmetria is frequently seen. The dysmetria is more apparent in the ipsilateral upper extremity. These patients also have a peculiar type of dysmetric eye movements that are characterized by hypermetric saccades toward the side of the lesion and hypometric saccades to the opposite side. We examined four patients with acute Wallenberg's syndrome and found horizontal dysmetria of the affected extremity. Hypermetric arm and hand movements were present to the side of the lesion and hypometric movements toward the opposite side. This type of dysmetria is probably related to the same patophysiological mechanism that underlies dysmetric eye movements in the Wallenberg's syndrome. The dysmetria tends to disappear with time although it continues to be present in some patients six months after the ischemic damage. PMID- 11061104 TI - [Cost-effectiveness analysis of ceftriaxone and cefotaxime in the treatment of community-acquired pneumonia]. AB - OBJECTIVE: To assess the cost-effectiveness ratio of ceftriaxone and cefotaxime to treat moderate to severe community acquired pneumonia (CAP). METHODS: A clinical trial was done in five hospitals of the Instituto Mexicano del Seguro Social, at the metropolitan area of Mexico City. Ceftriaxone and cefotaxime were compared to treat moderate to severe CAP, and the costs of purchasing, preparation, administration, hospitalization, and therapeutic success were quantified. Cost-effectiveness ratio was calculated, and sensitivity analysis and incremental analysis were done. RESULTS: The main isolated germs were Streptococcus pneumoniae (23.6%) and Staphylococcus aureus (18.5%). Most of the microorganisms were sensitive to ceftriaxone, ceftazidime, and cefotaxime, and were resistant to penicillin, ampicillin, and erythromycin. Therapeutic success was 98% in the ceftriaxone group and 83% in the cefotaxime group (p = 0.0091). Cost-effectiveness ratio for per cent unit of success was $19,458.62 Mexican pesos in the ceftriaxone group and $29,218.08 in the cefotaxime group. Sensitivity analysis showed consistently a lower cost-effectiveness ratio in the ceftriaxone group. Incremental analysis based on the treatment of 55 patients showed that using ceftriaxone instead of cefotaxime resulted in saving $35,170.79 per each additional cured patient. CONCLUSIONS: Ceftriaxone has a lower cost effectiveness ratio than cefotaxime to treat patients with CAP and bad prognosis criteria requiring hospitalization. PMID- 11061105 TI - Frequency of glutamic acid decarboxylase autoantibodies in Mexican diabetic children. AB - OBJECTIVE: Use radio binding assay (RBA) to quantify the frequency of autoantibodies to glutamic acid decarboxylase in Mexican children with type 1 diabetes mellitus (DM 1). RESEARCH DESIGN AND METHODS: GAD antibodies were measured in 140 mestizo children with DM 1, 66 female (47.14%) and 74 male (52.8%); age 11.7 +/- 3.55 years, and range 1.10 to 18.5 years. Most patients were treated with intermediate acting insulin, and some with the former combined with regular insulin. Mean disease duration was 3.11 +/- 2.94 years, and range 1 month to 14.5 years. Once the signed written consent was obtained, a 5.0-mL blood sample was drawn, immediately centrifuged, and the serum was kept frozen to -20 degrees C until RBA evaluation was performed with a commercial kit. RESULTS: The anti-GAD was positive in 76 DM 1 patients (54.28%) with values from 1.11 to 156.73 U/mL, and negative in 64 (45.71%). In 19 positive anti-GAD patients, the test was repeated and levels were found between 1.38 and 156.62 U/mL. An initial control group consisting of 25 healthy non-related volunteers matched by sex and age, showed negative anti-GAD for all. CONCLUSIONS: The frequency of anti-GAD in these patients was lower than that of the DM 1 European patients, but similar to that of Asians. This supports the heterogeneity of the etiopathogenic factors of DM 1 in different ethnic groups. PMID- 11061106 TI - [Physiology of erection and treatment of erectile dysfunction]. AB - Erectile dysfunction is a very prevalent disease, associated with chronic diseases mainly from the cardiovascular system. Better understanding of penile physiology has permitted the development of new drugs which offers each type of patient a specific and directed treatment. In the future, more basic and clinical research will surely result in better drugs. This article is intended to give insight to modern physiological concepts and apply them to a better understanding of therapeutic options available nowadays. PMID- 11061107 TI - [Genetic aspects of asthma]. AB - Asthma is a complex disease associated with bronchial hyperreactivity and atopy, making asthma a disease with a phenotype that has been clinically difficult to define. Despite intense research, prevalence of asthma remain relatively high. The key reason for the high prevalence and morbility is that the fundamental mechanisms predisposing individuals to the development of asthma are not understood. Familial aggregation observed in this pathology has prompted for the search of an involved genetic component. This task is difficult due to the complex nature of asthma. A universally accepted definition for this disease is not available, clinical expression can be modulated by environmental factors, and inheritance does not follow a clear Mendelian pattern. Establishment of more precise clinical and laboratory criteria has improved the design and interpretation of genetic studies. Twin analysis and segregation studies have demonstrated an important genetic component with a probably multifactorial pattern of inheritance. "Sib pair" studies and familial segregation analyses have shown linkage between some chromosomal regions and asthma, including chromosome 5, 6, 7, 11 and 14. The search for major genes in these chromosomal segments has been focused on loci involved in the allergic process. Among these, the loci for IL-9 and IL-13 in chromosome 5 seem to play an important role in the pathogenesis of asthma. Understanding the fundamental gene-environmental interactions in the development of asthma should lead to earlier identification of susceptible individuals and more effective approaches for disease prevention. PMID- 11061108 TI - [Crystal arthropathy]. PMID- 11061109 TI - [Role of genes of the major histocompatibility complex in infections]. PMID- 11061110 TI - [Hodgkin's disease and Epstein-Barr virus in Mexico; more evidence of their possible causal relationship]. PMID- 11061111 TI - [Diffuse lung calcinosis in a case or primary hyperparathyroidism]. PMID- 11061112 TI - [Generic drugs. Should they undergo clinical evaluation?]. PMID- 11061113 TI - [Contributions of the INCMNSZ to medicine: Thrombosis, antibodies, and systemic lupus erythematosus]. PMID- 11061114 TI - In search of oncology answers. PMID- 11061115 TI - The health care executive's view of management. PMID- 11061116 TI - Case identification and selection outcomes. PMID- 11061117 TI - The necessity of health information management. PMID- 11061118 TI - Building confidence in managed care. PMID- 11061119 TI - Positive outcomes: a case for rehab equipment specialists. PMID- 11061120 TI - Managed care is going down a bumpy road. PMID- 11061121 TI - Are you ready? Y2K compliance issues. PMID- 11061122 TI - Behavior adjustments in traumatic brain injury. A case study. AB - A traumatic assault to the brain often leaves significant residual effects, including severe behavioral dyscontrol. The person may demonstrate unwanted behaviors, such as physical aggression or verbal abuse, and lack motivation or the skills to engage in desirable behavior. ReMed, a community-based brain injury program, specializes in using a behavior analytic approach to address unwanted behaviors while understanding and developing strategies to further recovery from the brain injury. PMID- 11061123 TI - Payer-based case management: perspectives on managing brain-injured patient. AB - As the payer environment is carved into many segments, each with its own accountability relative to its financial liability, family members, providers, and even other case managers often find it difficult to comprehend the perspectives of the various parties represented. Indeed, one brain injury case I recently evaluated had four payer case managers: one from the patient's health plan, one from a disease management program for his premorbid diabetes, and two from Medicaid-sponsored programs for the disabled. For payer-based case managers, the ubiquitous case manager role confusion compounds product unfamiliarity. PMID- 11061124 TI - Functional rehabilitation of employees with chronic musculoskeletal pain. AB - Many patients with chronic musculoskeletal pain suffer associated functional limitations in work capacity and personal productivity. However, pain differs among patients and is quite difficult to measure. Although some validity lies in an individual's reports of pain, no generally accepted measurements are applicable between individuals. On a scale of 0 to 10, changes in pain that a patient reports are valid only with respect to that person. However, what constitutes one person's rating of 10 may represent only 5 or 6 to another person, so pain rating scales have only a limited role in assessing impairment. PMID- 11061125 TI - Chronic pain management: evaluating the use of opioids. AB - A patient with chronic pain who is on multiple medications raises important questions for the case manager. Is the patient's underlying problem actually pain, or is it addiction? Has the patient been thoroughly evaluated? Does the patient have a coordinated management plan, or are several physicians independently writing prescriptions and recommending treatment? How can the case manager facilitate appropriate management? PMID- 11061126 TI - Keys to managing oncology care. AB - Each year, more than 1 million Americans are diagnosed with some form of cancer. Although the term cancer includes more than 100 different diseases, half of all cancers can be successfully treated or cured if the disease is detected early and appropriate therapy is initiated and administered as scheduled with minimal interruptions and dose modifications. Because of higher cure rates and longer survival, the current approach is to view cancer as a chronic rather than terminal illness. PMID- 11061127 TI - Oncology clinical trials. AB - In 1971, President Nixon declared a "war on cancer" and initiated substantial funding for the National Cancer Program, which has been sustained through the years with a significant return on investment. Recently released 1998 statistics from the National Cancer Institute (NCI), the Centers for Disease Control and Prevention, and the American Cancer Society show the first real decline in cancer since the 1930s. Still, more than 1.2 million Americans will be diagnosed with cancer this year. New developments in biomedicine and advances in science and technology likely will lead to greater declines in cancer incidence and mortality. PMID- 11061128 TI - Oncology reimbursement changes on the horizon. AB - Did anyone ever say being a case manager was simple? Today's rapidly changing health care environment definitely has altered that perspective. Now more than ever, the case manager must be aware of the latest clinical interventions for the population of patients he or she serves and the financial picture painted by a variety of reimbursement sources. PMID- 11061129 TI - [Marie--a little miracle--views of a pregnant mother]. PMID- 11061130 TI - [The Asiatic infant]. PMID- 11061131 TI - [Psychosocial development of the infant during pregnancy, birth and early infancy]. PMID- 11061132 TI - [Pediatric gymnastic treatment possibilities in hip joint disorders in childhood and adolescence]. PMID- 11061133 TI - [Every night wide awake--sleeplessness in children]. PMID- 11061134 TI - [Methods of pain diagnosis and measurement of pain level in nursing practice. 2]. PMID- 11061135 TI - [Patients' rights in Germany today]. PMID- 11061136 TI - [The sick and healthy child, as painted by Otto Dix, from the viewpoint of a pediatrician]. PMID- 11061137 TI - [Survival strategies for gynecologic clinics and health policy reform measures from the viewpoint of the Society of Midwifes]. PMID- 11061138 TI - [Psychosomatic diseases in children and adolescents]. PMID- 11061139 TI - [Supine positioning of the sick child]. PMID- 11061140 TI - [Psychosocial conditions of drug-addicted mothers]. PMID- 11061141 TI - [Surgery on very small premature infants at the neonatal intensive care station]. PMID- 11061142 TI - [Children as visitors at a hospice]. PMID- 11061143 TI - [Speech development in children and speech disorders]. PMID- 11061144 TI - [He who criticizes me makes me wiser]. PMID- 11061145 TI - [Client orientation as a therapeutic objective at the clinic]. PMID- 11061146 TI - [Iodide uptake in children and adolescents in Germany--a situation analysis]. PMID- 11061147 TI - ["Laughter therapy with Dr. Troot, please!" Professional hospital-clowns brighten everyday life for hospitalized children]. PMID- 11061148 TI - [The Contergan (Thalidomide) case. Catastrophic consequences]. PMID- 11061149 TI - [Integrated basic training in pediatric and geriatric nursing]. PMID- 11061150 TI - [Chronic inflammatory intestinal diseases in childhood]. PMID- 11061151 TI - [Do we still need pediatric nurses?]. PMID- 11061152 TI - [Whooping cough in children and adults: SITKO (Permanent Inoculation Commission) draws conclusions]. PMID- 11061153 TI - [Definition of the Corporate Identity at the Intensive Care Station]. PMID- 11061154 TI - Regaining the public's trust. PMID- 11061155 TI - Associate degree graduates and the rapidly aging RN workforce. AB - This second segment of a four-part series examines the inter-relationship between the growth in associate degree nursing programs and the aging of the RN workforce. A growing proportion of new RNs have entered the workforce via associate degree programs, increasing from 40% in 1977 to 60% in 1996. New graduates, as well as working RNs, are approximately 5 years older in 1996 than 20 years earlier. Findings suggest that the rapid aging of the RN workforce can not be directly attributed to the rise in the number of older-aged graduates of associate degree programs. Rather, the declining propensity of those born after 1960 to enter nursing has resulted in fewer young RNs, and therefore: (1) an aging workforce, and (2) fewer new grads from baccalaureate programs (which have always attracted younger RNs) relative to grads from associate degree programs (which have always attracted older RNs). PMID- 11061156 TI - Alternative cost-effective anesthesia care teams. AB - Private and public payers are increasingly seeking an overall per-diem or global surgery rates that put hospitals at significant financial risk for anesthesia services. Other payers are demanding deep discounts in anesthesia fees and negotiating global capitation rates that put both hospitals and physicians at risk for all care including anesthesia. This study examines some of the various organizational models for using physician anesthesiologist (MDA) and nurse anesthetist (CRNA) resources most cost effectively and safely. Variations in percentages of these practitioners can be seen in that California reportedly has 47 MDAs for every 10 CRNAs while Michigan has just 6 MDAs for every 10 CRNAs practicing in that more highly managed care environment. Four various anesthesia practice models are described in detail without declaring any one a universal model. The cost per year for MDAs averages $294,000 while the cost per year for CRNAs is less than half as much. PMID- 11061157 TI - Ethics and economics. AB - The author suggests that one way to better manage the burgeoning costs in acute care settings and improve patient care is by the earlier use of ethics case consultations and end-of-life support from ethics teams. This study determined that, in several very diverse clinical scenarios, timely facilitation of meaningful communication and decision making between patients, families, and health care providers can result in the more appropriate use of health care resources. While few of the patients in this study had recorded advanced directives in place, and there was initially a lack of family consensus in some cases, compliance with the ethics team recommendations led to a more appropriate clinical unit placement; and improved family support helped manage the costs of care and focus on the patients' quality of life. The decrease in the use of medical interventions and therapies after ethics consultations was consistent in all cases presented here. PMID- 11061158 TI - Outcomes from use of an evidence-based practice guideline. AB - The aim of evidence-based guidelines is primarily to improve patient outcomes without adding to the existing cost of care because both payers and policymakers want to identify health care costs that do not result in benefit to the patient. The purpose of the reported project was to generate a practice guideline for the treatment of uncomplicated acute cystitis in a female population, to determine the extent to which the guideline would be used by providers and to measure the cost and quality of outcomes from its use. A retrospective chart review was used to gather pre-guideline practice and cost data. Measurements included the type, frequency, and duration of antibiotic therapy and the use of urine cultures and both complications and routine followup visits. The implementation of an outpatient practice guideline resulted in a significant change in antibiotic prescribing and a trend toward a change in ordering cultures and clinic followup. There was also a significant decrease in treatment costs. PMID- 11061159 TI - Disease management gains acceptance--and finds its legs--with automation. PMID- 11061160 TI - Making a play in the policy and politics arena: nursing and the national election. AB - With health care surfacing in political campaigns across the country and referencing virtually every one of the policy topics discussed here, nurses have an important opportunity to share their views regarding both policy substance as well as engage in the political arena. For those nurses not yet ready to run for elective office, participating in grassroots efforts of campaigns, on advisory panels, and making financial contributions to preferred candidates are important opportunities not to be missed. Given the interest in and challenges ahead associated with health care broadly and nursing specifically, there is a great deal at stake. PMID- 11061161 TI - The leader as a retention specialist. AB - Retention problems will plague us for the near future. Blanchard and Waghorn (1997) believe that everyone wants to be magnificent, and not just ordinary. Unfortunately, often we expect only the ordinary and not the magnificent. People respond to our expectations. We can help staff to achieve magnificence by providing unconditional love and remembering that people are magnificent. Sometimes their behavior is a problem and certain things block the expression of their greatness, but all people are magnificent. If we create "sanctuaries of caring" where nurses and others are given the opportunity to achieve their innate nobility and magnificence, people will flock to participate in the noble and magnificent experience of patient care. PMID- 11061162 TI - Integrated delivery systems: what's happening? PMID- 11061163 TI - The domain of telenursing: issues and prospects. PMID- 11061164 TI - Pay--the job's half done. PMID- 11061165 TI - Seize the moment. PMID- 11061166 TI - . . . and the penny dropped. PMID- 11061167 TI - Nursing against all odds. PMID- 11061168 TI - Under guarantee. PMID- 11061169 TI - Working it all out. PMID- 11061170 TI - Just a minute. PMID- 11061171 TI - Helpful people. PMID- 11061172 TI - Watching the clock. PMID- 11061173 TI - No workaholics need apply. PMID- 11061174 TI - Evidence-based nursing guidelines on the Web. PMID- 11061175 TI - Primary care groups: nurses on the board. AB - This article describes what happened in one region of England where 59 nurses who had put themselves forward were short-listed for appointment to 12 primary care groups. It identifies the leadership and motivational factors of those nurses and takes a retrospective view of the selection processes involved in appointing nurses to PCG boards in the region. PMID- 11061176 TI - Are we PREPared? AB - The author determined, by means of a small questionnaire survey carried out at a district general hospital, what understanding nurses have of post-registration education and practice (PREP) and whether they were prepared to meet the requirements set out by the United Kingdom Central Council for Nursing, Midwifery and Health Visiting (UKCC). His study also aimed to establish how Continuing Professional Development (CPD) requirements of staff at the hospital could be met. PMID- 11061177 TI - Screening for dementia and depression in older people. AB - Nurses have a valuable role to play in screening for dementia and depression in older people but they must be confident, adequately trained and supported. The authors conclude that screening and assessment is vital for older people to gain access to specialist mental health services. PMID- 11061178 TI - Pressure area care. PMID- 11061179 TI - Develop your career. PMID- 11061180 TI - Are you listening? PMID- 11061181 TI - A cyanobacterial recombination study, involving an efficient N2-fixing non heterocystous partner. AB - Many filamentous cyanobacteria fix atmospheric nitrogen under natural conditions in specialized anaerobic compartments, heterocysts, interspersed between vegetative cells, which provide protection to the O2-sensitive nitrogenase. A few unicellular cyanobacterial strains are also known to fix nitrogen aerobically at a slower rate. Filamentous cyanobacteria lacking heterocysts are not known so far to fix nitrogen. We describe the isolation and purification of a non heterocystous filamentous cyanobacterium from the fronds of the water-fern Azolla, fixing nitrogen at 18.7+/-0.2 n moles ethylene microg Chl. a(-1) h(-1) when grown in nitrogen-free medium at a low level of oxygen between two layers of agar. This strain of Anabaena azollae has been designated as het- nif+ (non heterocystous and nitrogen-fixing), and is found to be easily and effectively preserved in nitrogen-free medium in standard synthetic cyanobacterial nutrient medium (pH 8.5) at a continuous light intensity of 2800 lx at 25+/-1 degrees C. This het- nif+ strain is an effective donor of the nif+ marker to a het+ nif- strain of another cyanobacterium, Nostoc muscorum, when both are grown together in a recombination study. PMID- 11061182 TI - A study on the microbiota from olive-mill wastewater (OMW) disposal lagoons, with emphasis on filamentous fungi and their biodegradative potential. AB - The microbial composition of olive mill wastewater (OMW) from four disposal ponds has been studied. Such OMW samples contained a variable (but high) number of bacteria, yeasts and molds. Among the latest, members of twelve different genera (Acremonium, Alternaria, Aspergillus, Chalara, Fusarium, Lecytophora, Paecilomyces, Penicillium, Phoma, Phycomyces, Rhinocladiella and Scopulariopsis) were found. Members of five genera (Chalara, Fusarium, Paecilomyces, Penicillium and Scopulariopsis) were widely distributed, and they were able to grow efficiently in undiluted OMW as a sole source of nutrients. Strains of Fusarium, Paecilomyces, Penicillium and Scopulariopsis showed a marked capacity for OMW detoxification, depleting its antibacterial activity almost completely. PMID- 11061183 TI - Microflora of technogenous wastes characterised by fatty acid profiling. AB - Fatty acid methyl ester (FAME) profiles obtained directly in situ have been used to estimate microbial community structure in different technogenous wastes. The effect of nutrients added, simulating the effect of plant-derived exudates on the indigenous microflora in the heaps during the reclamation process, was also studied in microcosms. The wastes such as coal-mine spoil, non-ferrous metallurgical slag and coal fly-ash were characterised by a poorly developed microflora as compared to a typical sandy loam soil. However, the most similar to the soil was the community structure in the coalmine spoil. The high content of 18: 2omega6,9 found in the metallurgical slag indicated the domination of fungi in this waste. In contrast, representatives of the Cytophaga-Flavobacterium group dominated the coal fly-ash, for which 16:1omega5c was used as the marker acid. The waste amendment resulted in changes of FAME profiles obtained. However, the changes were site-specific, indicating response of particular microbial groups to the added nutrients. PMID- 11061184 TI - Isolation and characterization of two hydrocarbon-degrading Bacillus subtilis strains from oil contaminated soil of Kuwait. AB - Two strains of hydrocarbon-utilizing bacteria were isolated from soil samples of the Kuwait Burqan oil field at a temperature of 37 degrees C. The bacteria were motile endospore-forming rods with slight differences in their metabolic patterns and 16S rRNA sequence. Vegetative cells of the strains designated as AHI and AHII had an ultrastructure typical of gram-positive bacteria and showed gram-positive staining. The bacteria did not show pigmentation. Best growth was observed at 37 degrees C at neutral pH and NaCl concentrations in the range of 5-10 g per l. Both strains were obligatory aerobic and developed on synthetic media with either Diesel fuel, n-decan or naphthalene as the sole carbon and energy source. No specific growth factors were required. On the basis of their morphological, physiological and biochemical features, as well as their 16S rRNA analysis and electron microscope study, both strains were assigned to the species of Bacillus subtilis. PMID- 11061185 TI - Expression of soluble, recombinant alphavbeta3 integrin fragments in Escherichia coli. AB - No prokaryotic expression of integrin alphavbeta3 has been reported so far. We report here the expression of C-terminally truncated alphavbeta3 receptors in E. coli considering the known features required for dimerization and ligand binding. The expressed protein was insoluble despite of the addition of 'solubilizers' to the culture medium. Osmotic stress conditions combined with added exogenous solutes resulted in a small part of soluble receptor. The alphavbeta3 variants were purified from inclusion bodies or from soluble cytoplasmic maltose binding protein fusions. Heterodimerization of the subunits was proved by immunoprecipitation assays. Receptor-ligand binding was found to depend on the concentration. A competition assay with RGD peptides referred to unspecific receptor-ligand interaction. The latter fact was consistent with the finding that soluble receptors did not bind on RGD peptide-coupled sepharose (GRGDSPK sepharose). PMID- 11061186 TI - Reliable amplification of actin genes facilitates deep-level phylogeny. AB - The gene for actin as a highly conserved and functionally essential genetic element is developing into a major tool for phylogenetic analysis within a broad organismic range. We therefore propose a set of universally applicable primers that allow reliable amplification of actin genes. For primer construction the amino acid sequences of 57 actin genes comprising fungi, animals, plants and protists were analysed, aligned and used for the definition of six well-conserved regions which are suitable as priming sites in PCR amplification experiments. Ten primers were designed for specific in vitro amplification of actin gene fragments from a wide range of microorganisms. The corresponding gene fragments provide a strong basis to isolate nearly complete actin genes for further molecular characterization and for establishing phylogenies based on actin gene trees. PMID- 11061187 TI - An immunoassay for detection of heat-stable proteases from thermoduric psychrotrophic Bacillus spp. of dairy origin. AB - A homogeneous preparation of a thermostable protease from Bacillus sp. B-17 was used to raise an antiserum in rabbits. IgG of this antiserum was used to study the antigenic relationship of proteases in cell-free extracts of 21 bacilli of milk origin. Based on immunological cross reactivity, the 21 bacilli were divided into 3 serological subgroups. To raise antibodies of broader specificity, protease from Bacillus sp. B-11 (group II) and B-3 (group III) were purified, mixed with purified B-17 protease, and an antiserum was raised against this mixture. IgG of this antiserum was purified (IgG anti-bacilli protease). A sandwich ELISA was standardized using IgG anti-bacilli protease as capture antibody. The assay could detect 1.2 ng ml(-1) of protease in milk or buffer, but the assay failed to detect 4 of 21 bacilli proteases. The results suggest that this assay is useful for the detection of proteases of Bacillus spp. in dairy industry. PMID- 11061188 TI - Growth and survival of phosphate-solubilizing bacteria in calcium alginate. AB - Calcium alginate was superior to conventional charcoal-soil (3:1) carrier for phosphate-solubilizing bacteria (PSB). High populations of Pseudomonas striata (27) and Bacillus polymyxa (H-5) could be maintained in this polymer during storage. Incorporation of charcoal-soil (3:1) adversely affected the initial loading of these organisms in alginate gel. Alginate alone supported maximum survival of these organisms at elevated storage temperature (40 degrees C). PMID- 11061189 TI - Effect of water-soluble vitamins on the production of indole-3-acetic acid by Azospirillum brasilense. AB - The effects of six water-soluble vitamins on tryptophan-dependent synthesis of indole-3-acetic acid in Azospirillum brasilense were investigated. A multifactorial regression analysis was employed to produce models of indole-3 acetic acid synthesis versus concentrations of tryptophan and the vitamins added to the growth medium. Very low levels of the B-group vitamins added at 10 to 100 microg l(-1) affected production of indole-3-acetic acid in A. brasilense. The largest release of this phytohormone was observed after amendment with pyridoxine and nicotinic acid. Results of the study suggest a role these vitamins may fulfil in the regulation of indole-3-acetic acid synthesis in A. brasilense. PMID- 11061190 TI - Influence of salinity on methanogenesis and associated microflora in tropical rice soils. AB - In a laboratory incubation study, methane (CH4) production in two saline soils and a nonsaline soil sample was investigated under flooded conditions. Mean CH4 production was remarkable (630.86 ng CH4/g) in nonsaline alluvial soil, but low (12.97 ng CH4/g) in acid sulfate saline (Pokkali) soil which was attributed to the high sulfate content of the later. CH4 production was also low in the coastal saline (Canning) soil (142.36 ng CH4/g) but increased upon leaching the soil of its salt content. Addition of salts to the nonsaline alluvial soil at 4, 8, 16 and 20 dS/m progressively decreased CH4 production. The inhibition of CH4 production was related to low microbial activities as reflected by decreased microbial biomass C and low soil microbial population including that of methanogens. PMID- 11061191 TI - Impact of phosphate-solubilizing fungi on the yield and phosphorus-uptake by wheat and faba bean plants. AB - Three fungal isolates (phosphate-dissolvers), Aspergillus niger, A. fumigatus and Penicillium pinophilum were isolated from the rhizosphere of different plants grown in Ismailia and South Sinai Governorates. They effectively solubilized rock phosphate or tricalcium phosphate in Pikovskaya's liquid medium. In pot and column experiments, they significantly reduced pH and increased available phosphorus in the soil treated with either rock phosphate or superphosphate. The yield components of wheat and faba bean plants increased as a result of soil inoculation with the isolated fungi. Penicillium pinophilum was the most efficient isolate. It increased the yield of wheat grains by 28.9 and 32.8% in the soil treated with rock phosphate and superphosphate, respectively. Similarly, it increased the production of faba bean seeds by 14.7 and 29.4% with the same treatments. The uptake of phosphorus by both crops significantly increased due to inoculation of the soil with the tested fungi. PMID- 11061192 TI - Bacterial endobionts in the big non-mycorrhizal roots of Scots pine (Pinus sylvestris L.). AB - Subsurface bacterial growth occurred in an N-free medium inoculated with interior tissues of big non-mycorrhizal roots (7 to 8 mm diameter) of 15-20 years-old Scots pine (Pinus sylvestris L.) growing on sand dunes at the Baltic Sea of Poland. The bacteria were not N2 fixers as determined by the acetylene reduction method. Light microscopic and scanning electron microscopic observations revealed massive bacterial clusters residing in the cortical cells underlying epidermis and parenchyma. The bacteria produced yellow-green pigments on King's medium, which fluoresced under ultraviolet (UV) irradiation at 366 nm wavelength, and could be a siderophore-producing Pseudomonas. PMID- 11061193 TI - Antifungal characteristics of a fluorescent Pseudomonas strain involved in the biological control of Rhizoctonia solani. AB - A plant growth-promoting isolate of a fluorescent Pseudomonas spp. EM85 was found strongly antagonistic to Rhizoctonia solani, a causal agent of damping-off of cotton. The isolate produced HCN (HCN+), siderophore (Sid+), fluorescent pigments (Flu+) and antifungal antibiotics (Afa+). Tn5::lacZ mutagenesis of isolate EM85 resulted in the production of a series of mutants with altered production of HCN, siderophore, fluorescent pigments and antifungal antibiotics. Characterisation of these mutants revealed that the fluorescent pigment produced in PDA and the siderophore produced in CAS agar were not the same. Afa- and Flu- mutants had a smaller inhibition zone when grown with Rhizoctonia solani than the EM85 wild type. Sid- and HCN mutants failed to inhibit the pathogen in vitro. In a pot experiment, mutants deficient in HCN and siderophore production could suppress the damping-off disease by 52%. However, mutants deficient in fluorescent pigments and antifungal antibiotics failed to reduce the disease severity. Treatments with mutants that produced enhanced amounts of fluorescent pigments and antibiotics compared with EM85 wild type, exhibited an increase in biocontrol efficiency. Monitoring of the mutants in the rhizosphere using the lacZ marker showed identical proliferation of mutants and wild type. Purified antifungal compounds (fluorescent pigment and antibiotic) also inhibited the fungus appreciably in a TLC bioassay. Thus, the results indicate that fluorescent pigment and antifungal antibiotic of the fluorescent Pseudomonas spp. EM85 might be involved in the biological suppression of Rhizoctonia-induced damping-off of cotton. PMID- 11061194 TI - The effect of benomyl on growth and ultrastructure of two isolates of Phytophthora infestans from Egypt. AB - The effect of benomyl as a fungicide on the growth rate and ultrastructure of two isolates (P1319 and P623) of Phytophthora infestans is compared. Benomyl caused a concentration-dependent inhibition of the mycelial growth of both isolates. The isolate P1319 was found to be more sensitive to benomyl than the isolate P623. Ultarstructural studies confirmed these observations. The hyphae of isolate P1319 subjected to 100 and 500 ppm benomyl showed more severe changes in the cytoplasm than those of isolate P623. An increase in lipid bodies and vacuoles in the hyphal cytoplasm was the characteristic phenomenon after treatment with benomyl, particularly at a concentration of 500 ppm. PMID- 11061195 TI - Amputees in Limburg: incidence, morbidity and mortality, prosthetic supply, care utilisation and functional level after one year. AB - OBJECTIVES: To describe for the province of Limburg, the Netherlands, the incidence of lower limb amputation (LLA), the demographic characteristics of the amputees, reason and level of amputation, care the amputees receive, prosthetic supply and functional level after one year. DESIGN: A prospective descriptive cohort study. METHODS: Anonymized data on all amputees in all nine hospitals were collected during 1994. Follow-up was one year. RESULTS: 191 major lower limb amputations in 164 patients. Incidence was 17.1 per 100,000. Amputation levels: 77 trans-tibial, 52 transfemoral and 43 knee disarticulation. Twenty (20) patients died during hospitalisation. Eighty-seven (87) amputees gave permission for followup, 60 could be actually followed. Twenty-two (22) patients received in patient rehabilitation, 16 primary day-care, 21 went to a nursing home and one went home without rehabilitation. The mean duration of rehabilitation was 35 weeks. In 53 patients a functional prosthesis was indicated. Forty-three (43) of these patients walked with the prosthesis after one year, 7 of whom more than 500 metres. Nineteen (19) amputees performed the Get Up and Go Test (GUGT) safely. Amputees on average have a low level of functioning, as indicated by SIP68 and Barthel Index Scores. The ability to walk is closely related to this daily function and quality of life. CONCLUSION: After one year the majority of amputees have low walking skills and the walking distance is limited. They are often ADL dependent and their amputation greatly limits their daily function. PMID- 11061196 TI - Developments in the trans-tibial prosthetic socket fitting process: a review of past and present research. AB - A revolution in trans-tibial prosthetic design began at the end of World War II with the development of new materials and a dramatic improvement in the understanding of biomechanics. Early research was based mainly on the improvement of existing prosthetic design practice. Today, research has been focused on providing a better understanding of stump/socket interface biomechanics and improving socket fit by attempting to quantify the normal/direct stresses at the interface. The purpose of this review paper is to question whether research and prosthetic education/training to date has significantly improved our understanding of what makes a good socket. Although there is no doubt that advances in socket fitting techniques have been made what is not clear is the actual extent to which these advances have improved the quality of sockets fitted. It is suggested that a new approach is needed which can overcome some of the inherent problems of designing and manufacturing a comfortable high quality socket. It is also suggested that current research and education/training in the fields of pressure/interfacial interaction measurement and Finite Element Analysis techniques have limited potential to address many of these problems. There is also little evidence that current computer aided design systems offer any significant advantages over more conventional techniques. PMID- 11061197 TI - Equilibrium and movement control strategies in trans-tibial amputees. AB - This study was aimed at identifying changes in equilibrium and movement control strategies in trans-tibial amputees (TTA) related to both the biomechanical changes and the loss of afferent inflow. The coordinations between equilibrium and movement were studied in traumatical TTA and in controls during transition from bipedal to monopodal stance. TTA failed to perform the task in a high percentage of trials both when the sound and the prosthetic limb were supporting. Significant differences were also found between TTA and controls in the duration of the weight transfer phase, in the length of the initial centre of pressure (CP) displacement and in the electromyographic (EMG) patterns. Despite adaptive posturomotor control strategies, transition from bipedal to monopodal stance remains a difficult task to perform for TTA, both when the supporting limb is the affected one and when the sound one is. The results of this study are discussed with respect to the rehabilitation programme and the prosthesis design for transtibial amputees. PMID- 11061198 TI - The functional demands on the intact limb during walking for active trans-femoral and trans-tibial amputees. AB - The aim of this study was to investigate the loading demands placed on the intact limb in terms of joint moments and power for active trans-femoral and trans tibial amputees in comparison to a group of able-bodied subjects. Four (4) trans tibial, 4 trans-femoral amputees and 10 able-bodied subjects walked at 1.2m.s(-1) along a walkway whilst kinematic data from both the intact and prosthetic limbs, and kinetic data from the intact limb only were collected. A Panasonic VHS video camera was used to film subjects walking in the sagittal plane with simultaneous force data collected from a Kistler force platform. The amputees were found to compensate for the functional loss of one or more joints by increasing net joint moments and power output on their intact limb compared to able-bodied subjects. At the intact limb ankle, the range of motion, peak dorsiflexor moment and power generation at toe-off increased. At the intact limb knee, power generation during stance and extensor moments and power absorption at toe-off increased. At the intact limb hip, extensor moment and power absorption during stance, and hip flexor moment and power generation at toe-off increased. These findings were partly attributed to the prostheses used but mainly to adaptation mechanisms displayed by trans-femoral and trans-tibial amputees. They have implications for the mobility of amputees and the long term health of their joints. It was recommended that prosthesis design, prosthesis fitting and training in the use of the prosthesis were all factors which could be investigated with a view to minimising intact limb loading. PMID- 11061199 TI - Lower limb prosthesis utilisation by elderly amputees. AB - The goal of prosthetic rehabilitation is to compensate for the loss of a limb by amputation by, in the case of a lower limb, encouraging walking, and to achieve the same level of autonomy as prior to the amputation. However, because of difficulties walking, elderly amputees may use their prosthesis to a greater or lesser degree or simply stop using it during the rehabilitation period. The objective of this research was to study factors such as physical and mental health, rehabilitation, physical independence and satisfaction with the prosthesis to understand why amputees use their prosthesis or not. The sample was composed of 65 unilateral vascular amputees 60 years old or over living at home. The information was collected from medical records, by telephone interview and by mail questionnaire. Prosthesis use was measured by a questionnaire on amputee activities developed by Day (1981). Eighty-one per cent (81%) of the subjects wore their prosthesis every day and 89% of this group wore it 6 hours or more per day. Less use of the prosthesis was significantly related to age, female gender, possession of a wheelchair, level of physical disability, cognitive impairment, poorer self-perceived health and the amputee's dissatisfaction. A multiple regression analysis showed that satisfaction, not possessing a wheelchair and cognitive integrity explained 46% of the variance in prosthesis use. PMID- 11061200 TI - Evaluation of the psychophysical detection threshold level for vibrotactile and pressure stimulation of prosthetic limbs using bone anchorage or soft tissue support. AB - In the present study the psychophysical detection threshold levels mechanical stimulation of 32 prosthetic limbs were determined. Prosthetic limbs were anchored to the bone by means of an implant (n=17) or supported by a socket enclosing the amputation stump (n=15). Detection threshold levels were assessed for pressure and vibratory stimulation of the prosthesis and the limb at the contralateral side (control). Following vibratory stimulation, thresholds were increased on an average 20% for socket prostheses. but approached those of the control for bone-anchored prostheses. For pressure stimulation, thresholds were increased up to 60% for socket prostheses and 40% for bone-anchored prostheses compared to the control. While bone-anchored prostheses yielded significantly lower threshold levels than socket prostheses, there was no significant difference between both treatments regarding pressure stimulation. Results were applicable to both upper and lower limb amputees. It could be concluded that detection thresholds for pressure and especially vibratory stimulation of prosthetic limbs were generally higher than for control limbs. The outcome was related to the prosthetic limb design with bone-anchored prostheses yielding better perception than socket prostheses. PMID- 11061201 TI - Factors conditioning the return to work of upper limb amputees in Asturias, Spain. AB - Reintegration into a social and work environment, as the final objective rehabilitation therapy, is one of the greatest challenges faced in this speciality. The aim of this study was to analyse the reintegration into the workforce of 43 upperlimb amputees in Asturias, Spain (1,100,000 inhabitants) whose amputations were as a result of accidents at work. For this purpose various factors related to their return to work were studied. The most important factor was the year in which the amputation was carried out, since reintegration was more likely to occur in those amputees whose accident at work took place before the 1980s. PMID- 11061202 TI - Effectiveness and biomechanics of spinal orthoses in the treatment of adolescent idiopathic scoliosis (AIS). AB - In this prospective study, the effectiveness and biomechanical factors of spinal orthoses in the treatment of moderate adolescent idiopathic scoliosis (AIS) patients were investigated. In the first 20 months of orthotic treatment, the values of standing AP Cobb's angle, apical vertebral rotation, lumbar lordosis as well as thoracic kyphosis showed significant reduction (P<0.05), however, the angle of trunk inclination and trunk listing did not. The values of those reducible parameters reached their lowest values within the first 12 months of orthotic treatment and then the values gradually increased but they were still below the pre-brace values. The mean pressure of The pressure pads was found to be 7.09 +/- 1.77 kPa (53.2 +/- 13.3 mmHg) while the mean tension of the straps was 26.8 +/- 5.2N. The standing AP Cobb's angle strongly correlated with the pad pressure (correlation coefficient=0.931, p<(.05) and strap tension (correlation coefficient=0.914, p<0.05). The strap tension and pad pressure strongly correlated and the correlation coefficient was 0.873 (p<0.05). This suggests that in the consideration of biomechanical function of spinal orthoses, the focus may be upon how tightly the orthosis was fastened and if the location and direction of the pressure pads are the correct. Therefore, for enhancing independent standard tension should be set in each strap, and regular and close monitoring is needed. PMID- 11061203 TI - Changes in interface pressure and stump shape over time: preliminary results from a trans-tibial amputee subject. AB - Interface stresses and stump shape were measured during sessions over a two-month interval on a trans-tibial amputee subject. Results from thirteen transducer sites monitored during four sessions showed greater interface pressure changes over time at anterior sites than at lateral or posterior locations. There was a trend of decreased pressure with stump swelling and increased pressure for stump atrophy. During one session in which stump shape was monitored over a 23.1 min interval after ambulation, stump swelling was localised. Swelling tended to increase in the regions of initial enlargement, as opposed to redistributing through different areas over time. Regions of swelling were anterior lateral and posterior proximal, areas of thick underlying soft tissue. Identification of localised areas of swelling and atrophy and understanding of their effects on interface pressures could be used to improve individual socket design. PMID- 11061204 TI - Non-surgical options to autologous finger transplants. AB - This case report describes the rehabilitation path of a man who sustained severe burns to his body and extremities as a result of a bushfire. The fingers of both hands were amputated except the thumbs and although he was a candidate for autologous transplant, declined this option for various reasons. Bilateral prostheses were made and resulted in a significant improvement in functional outcome without the need for surgery. Prostheses are practical alternatives to surgery and should be considered in selected patients. PMID- 11061205 TI - The relation between foetor ex ore, oral hygiene and periodontal disease. AB - Bad breath usually originates in the mouth. It is described with different names as oral malodor, halitosis or foetor ex ore. Dental plaque, bacterial products from deep periodontal pockets and bacterial products from the tongue probably cause bad breath but also bacterial products from tonsils and pharynx probably are involved. In this study we clinically examined subjects with very strong bad breath, foetor ex ore. Foetor ex ore was defined as strong evil-smelling odor from the mouth of the patient which had an affect on the examiner and made the oral examination excruciating. Subjects with foetor ex ore are not aware of it. It is usually noticed by others. There are also persons who complain of bad breath that cannot be detected by others, halitophobia. Our aim was to study the relation between foetor ex ore, halitophobia and oral hygiene, periodontal disease. A total of 840 men, mean age 35.7(+/- 2.8 SD) and 841 women, mean age 35.7+/- 2.9 SD), participated. Clinical findings were noted, including the presence or absence of foetor ex ore. The subjects also filled in a self-reported questionnaire concerning problems in the oral cavity and teeth. Foetor ex ore was present in 2.4 percent of the subjects. Multiple regression analysis showed that calculus (P < 0.001), plaque (P < 0.01), and dental visits once every 3 yr. (P < 0.01) were significantly correlated to foetor ex ore. Periodontitis patients with foetor ex ore had more severe disease (P < 0.001) than those without. Foetor ex ore was not related to suspected halitosis. One percent of the subjects had suspected halitosis. Using multiple regression analysis, we found a significant correlation between calculus (P < 0.001) and suspected halitosis. In conclusion this study shows that foetor ex ore was correlated to oral hygiene and dental visits. Periodontitis patients with foetor ex ore had more severe disease than those without. PMID- 11061206 TI - Temporomandibular disorders in juvenile chronic arthritis patients. A clinical study. AB - In a prospective study, 105 children with juvenile chronic arthritis (JCA) were clinically and radiographically examined. The aim was to investigate the distribution of symptoms and clinical signs of temporomandibular disorders and to study correlations with radiographic mandibular condylar lesions. The present material appeared to be a representative sample of Swedish JCA children with respect to the distribution of genders and JCA subtypes as well as the peak of onset of the disease. Symptoms from the masticatory system were common (26%) and pain at mandibular function and stiffness at mouth opening the most frequent ones. Restricted maximal voluntary mouth opening (MVM) was the most frequent clinical finding. Radiographic condylar lesions, frequently found in the present material (39%), were significantly correlated with postnormal occlusion, restricted MVM, anterior open bite and mandibular retrognathia. However, most of the JCA children with radiographic condylar lesions did not present with symptoms from the masticatory system, postnormal occlusion, restricted MVM, anterior open bite or mandibular retrognathia. PMID- 11061207 TI - Fixed prosthodontics in adults in Jonkoping, Sweden in 1983 and 1993. An epidemiological study of prevalence and choice of material. AB - The aim of the study was to assess the prevalence of fixed prostheses, i.e. single crowns and fixed partial dentures, in adults 20-80 years old in two cross sectional studies carried out in 1983 and 1993 and to analyse whether the choice of material for fixed prostheses had changed during this time period. The material comprised 586 and 593 individuals. A descriptive analysis of number of individuals with fixed prostheses was made concerning the number of crowns and pontics, the distribution in the jaws, and the materials used. The prevalence of individuals with fixed prostheses was shown to increase with age both in 1983 and 1993. In 1993 the number of individuals with fixed prostheses was slightly lower than in 1983 (44 and 48 percent respectively). There was no difference according to gender. The proportion of individuals with fixed prostheses was lower or almost unchanged in the 20-, 30-, 40-, 50-, 60-, and 80-year-olds in 1993 compared to 1983. In the 70-year-olds, however, the proportion of individuals with fixed prostheses was considerably higher in 1993 compared to 1983. Most individuals with fixed prostheses had a small number of crowned teeth, and about half of the individuals had not more than four crowned teeth. Likewise most individuals with pontics had a small number of pontics. The distributions of crowned teeth and pontics in the jaws were similar in 1983 and 1993. Pontics and crowns were more frequent in the upper than in the lower jaw. Crowns made of porcelain or metal ceramic had increased by 1993 in the 30-80 year age groups, and the study thus confirms general clinical experience that porcelain and metal ceramic are more often the materials of choice in fixed prosthodontics in adults today. PMID- 11061208 TI - Ability to estimate oral health status and treatment need in elderly receiving home nursing--a comparison between a dental hygienist and a dentist. AB - The aim of this study was to compare the estimation ability of a dental hygienist to that of a dentist when, independently, recording the oral health status and treatment need in a population of elderly, receiving home nursing. Seventy-three persons, enrolled in a home nursing long-time care programme, were recruited. For the oral examination a newly developed protocol with comparatively blunt measurement variables was used. The oral examination protocol was tested for construct validity and for internal consistency reliability. Statistical analyses were performed using Wilcoxon matched pairs signed rank sum test for testing differences, while inter-examiner agreement was estimated by calculating the kappa-values. Comparing the two examiners, good agreement was demonstrated for all mucosal recordings, colour, form, wounds, blisters, mucosal index, and for the palatal but not the lingual mucosa. For the latter, the dental hygienist recorded significantly more changes. The dental hygienist also recorded significantly higher plaque index values. Also regarding treatment intention and treatment need, the dental hygienist's estimation was somewhat higher. In conclusion, when comparing the dental hygienist's and the dentist's ability to estimate oral health status, treatment intention, and treatment need, some differences were observed, the dental hygienist tending to register "on the safe side", calling attention to the importance of inter-examiner calibration. However, for practical purpose the inter-examiner agreement was acceptable, constituting a promising basis for future out-reach activities. PMID- 11061209 TI - Fluoride release in vitro from a resin-modified glass ionomer after exposure to NaF solutions and toothpastes. AB - The aims of the present investigation were: (i) to study the release of fluoride from a resin-modified glass ionomer cement (Vitremer) after exposing ("recharging") the material with NaF toothpastes and NaF solutions with different fluoride concentrations, and (ii) to study the effect of covering the material with a sealant layer (glaze) in this respect. Totally 160 specimens were made, which were placed in water for 13 weeks to receive a low fluoride release value. The specimens were then randomly divided into 10 groups with 16 discs each. Five of the groups were exposed once a day for 7 days to one of the following fluoride agents: 0.05, 0.2 or 2% NaF solutions and slurries of two NaF toothpastes (Acta and Pepsodent). The other five groups received the same treatment twice a day. All treatments had a recharging effect. However, the 2% NaF solution resulted in significantly more fluoride release than the other solutions and toothpastes. Treatment twice a day gave higher total release than once a day, but the difference were only significant for the 0.2 and 2% NaF solutions. With glaze material covering the specimens, almost no fluoride release was observed. However, when the glaze was removed, a burst of fluoride occurred. PMID- 11061210 TI - Is semicarbazide-sensitive amine oxidase in blood plasma partly derived from the skeleton? AB - Semicarbazide-sensitive amine oxidases (SSAOs) are widely expressed copper containing enzymes. One enzyme of this family have high specific activity towards benzylamine and is present in human blood plasma. This enzyme is altered in several diseases, for instance in diabetes. Presently it is unclear where the plasma SSAO is synthesized. Previous autoradiographic studies have suggested that SSAO may be expressed in bone tissue. In the current study we have analyzed levels of SSAO in serum from cases with 'skeletal disease', i.e. patients with severe skeletal metastases of prostate cancer and subjects having recent fractures. Interestingly, subjects with metastases showed significantly elevated levels of SSAO in serum compared to individuals having prostate cancer without skeletal metastases. It is speculated that, at least in part, SSAO in the blood stream may be derived from bone tissue. PMID- 11061211 TI - Modification of dopamine release by selective inhibitors of MAO-B. AB - Chronic low dose deprenyl treatment in rats causes an increase in striatal extracellular dopamine level, without significant reduction in deaminated metabolite formation. This effect could be the result of increased endogenous levels of the MAO-B substrate beta-phenylethylamine, which is both a releaser of dopamine as well as an inhibitor of the neuronal membrane active dopamine uptake. In guinea pigs, however, striatal extracellular dopamine was not increased either by deprenyl or by clorgyline. Local infusion of the dopamine uptake inhibitor GBR 12909 caused a greater increase in striatal dopamine in microdialysate in rats than in guinea pigs. Intra-species differences in synaptic architecture or in density of dopamine transporter expression may account for these differences. PMID- 11061212 TI - Transdermal formulations of deprenyl: guinea pig and pig models. AB - The efficacy of many drugs relies on their presence at the site of action over a period of time. The retardation or programmed release capability of the conventional dosage forms like oral and parenteral are limited and toxic and undesired side-effects may occur after their applications. These problems may be solved using transdermal delivery systems. Transdermal systems are aimed for local, or systemic action. In the letter case controlling the rate of delivery or modulating the distribution in the organism. The selection of an adequate biological method of evaluating a new transdermal formulation is a critical point of the development. The in vitro methods can help in the characterization of the different formulas, but without an in vivo disposition study they cannot give relevant information about the expectable therapeutic behavior. We adapted and improved an in vivo test system for the evaluation of new transdermal particulate systems (patches) and liposomes containing deprenyl selegiline as active ingredient. The in vivo evaluation system consists of two steps: 1. Full biodisposition study on guinea pig, using isotope labeled selegiline. 2. Biodisposition studies on domestic pigs including dose, area, surface dependence and comparative bioavailability with traditional dosage forms and application moods. Specific examples of these studies and experimental technology are presented. PMID- 11061213 TI - An autoradiographic method of visualising semicarbazide-sensitive amine oxidase activity in mouse tissue sections. AB - Under the influence of semicarbazide-sensitive amine oxidase (SSAO), methylamine is deaminated to formaldehyde, which can react with various macromolecules and form irreversible adducts. We hereby present an autoradiographic method of visualising SSAO activity by measuring the in vivo formation of such adducts from 14C-methylamine. Our results revealed high concentrations of radioactive deposits in the intestinal wall, brown adipose tissue, spleen and bone marrow. Hydralazine is a potent SSAO inhibitor and pretreatment with this irreversible inactivator resulted in a nearly complete loss of radioactive deposits in the tissues. By giving 14C-methylamine at different time-points after irreversible inhibition of SSAO, it was also possible to determine the resynthesis rate of SSAO. Interestingly, the recovery rate of SSAO after such inactivation was tissue specific. The possible therapeutic value of a specific SSAO inhibitory drug has been discussed. PMID- 11061214 TI - (-)Deprenyl (Selegiline): past, present and future. AB - (-)Deprenyl (Selegiline), the N-propargyl analogue of (-)methamphetamine, is the only drug in clinical case which, by enhancing the impulse propagation mediated release of noradrenaline and dopamine in the brain (catecholaminergic activity enhancer, CAE, effect), keeps in small doses without side-effects the catecholaminergic brain system on a higher activity level. (-)Deprenyl stimulates the catecholaminergic neurons selectively in the brain because, in contrast to PEA and the amphetamines which induce the continuous release of noradrenaline and dopamine from their intraneuronal stores, (-)deprenyl is devoid of this property. It is due to the CAE effect that a) the maintenance of rats on (-)deprenyl during the postdevelopmental phase of their life slows the age-related decline of sexual and learning performances and prolongs life significantly; b) patients with early, untreated Parkinson's disease maintained on (-)deprenyl need levodopa significantly later than their placebo-treated peers, and when on levodopa plus ( )deprenyl, they live significantly longer than patients on levodopa alone; and c) in patients with moderately severe impairment from Alzheimer's disease, treatment with (-)deprenyl slows the progression of the disease. It is reasonable to expect that a prophylactic low dose administration of a safe catecholaminergic activity enhancer substance during the postdevelopmental phase of life will slow the age related decline of behavioral performances, delay natural death and decrease susceptibility to Parkinson's disease and Alzheimer's disease. PMID- 11061215 TI - Selective inhibitors and computer modelling of the active site of monoamine oxidase. AB - MAO inhibitors can be employed for computer modelling of the active site of MAO A and B. Competitive fully reversible MAO inhibitors with rigid structure and limited number of conformers are preferential compounds for these studies. Among various isatin analogues with nearllanar structure selective MAO B inhibitors fit to 3D box of 8.5x5.1x1.8 A, whereas 3D box of 14.2x5.6x1.8 A accommodates selective MAO A inhibitors. Validity of these data was tested using a series of pyrazinocarbazoles, analogues of short-acting antidepressant pirlindole. Rigid analogues exhibiting potent and selective inhibition of MAO A have 3D size limits of 13x7x4.4 A. Flexible analogues also demonstrated potent inhibition of MAO B and in contrast to rigid analogues their inhibitory activity did not show any dependence on 3D sizes. 3D-QSAR with CoMFA of isatin and pirlindole analogues of MAO A and B revealed differences in the models of MAO A and B. PMID- 11061216 TI - Inhibitor sensitivity of human serum and vascular semicarbazide-sensitive amine oxidases. AB - Recent data suggest that elevated serum semicarbazide-sensitive amine oxidase activity (SSAO) may cause endothelial injury. Formation of cytotoxic metabolites (especially formaldehyde) and increased oxidative stress might lead to initiation or progression of atherosclerosis. Effective and selective inhibitors of human SSAO might exert cytoprotective effect on endothelial cells. To compare the inhibitor sensitivity of human serum and vascular tissue SSAO enzyme, the inhibitory effect of semicarbazide and MDL 72974A was investigated. Serum and vascular SSAO activity has been determined using 14C-benzylamine as a substrate. The IC50 values of semicarbazide were estimated to be 5x10(-3) M and 5x10(-4) M for SSAO from human serum and saphenous vein, respectively. MDL 72974A amine oxidase inhibitor was more than thousand times more effective than semicarbazide. The IC50 values were 10(-7) M and 10(-8) M for SSAO from human serum and saphenous vein, respectively. This finding supports the hypothesis that soluble and membrane-bound vascular SSAO enzymes might have similar structure. PMID- 11061217 TI - The vulnerable coronary plaque. AB - Vulnerable coronary plaques are asymptomatic atherosclerotic lesions with the tendency to rupture. Plaque rupture is the initiating event in most acute coronary syndromes including sudden cardiac death, acute myocardial infarction, and unstable angina. Vulnerable plaques are commonly found in coronary arteries at autopsy but are virtually undetectable by standard diagnostic techniques such as stress testing and coronary angiography. Using new imaging techniques, in particular intravascular ultrasound and magnetic resonance imaging (MRI), scientists are now able to identify these plaques in vivo. A better understanding of the pathophysiology of plaque vulnerability and rupture will eventually lead to the therapeutic goal of plaque stabilization in the prevention of acute coronary syndromes. This article reviews the role of plaque vulnerability in coronary artery disease. The anatomy and pathophysiology of vulnerable plaques as well as diagnostic and therapeutic implication will be described. PMID- 11061218 TI - Inflammation and infection in acute coronary syndrome. AB - Basic science research has revealed that monocytes and macrophages are important factors in atherogenesis. Immune system activation occurs at all stages of plaque formation, from the fatty streak to an advanced, complicated lesion. The inflammatory response not only stimulates changes in coronary artery endothelial cells causing endothelial injury and dysfunction, but also plays a role in plaque instability and rupture. New perspectives of atherosclerosis and acute coronary syndromes will be discussed in relation to inflammation. In addition, discussion will focus on bacterial and viral infectious microorganisms as a potential factor that may induce and promote inflammation and lead to acute coronary events. Clinical studies in humans have provided insight relating inflammation and infectious agents to atherosclerosis and plaque vulnerability. Other studies focus on specific interventions that may aid in diagnosis and treatment. PMID- 11061219 TI - Surgical management of unstable angina and symptomatic coronary artery disease. AB - The treatment of coronary artery disease and, in particular, acute coronary syndromes has evolved from watchful waiting to an early aggressive intervention strategy. Patients are currently receiving either percutaneous or surgical revascularization. Several major clinical trials have identified those patients mostly likely to benefit from surgical intervention. These patients typically include those with left-main coronary artery disease, triple vessel disease with decreased left ventricular function, and other clinical risk factors. As a result of these studies, unique needs and outcomes of special populations have been identified. This article will present an overview of surgical treatment of coronary artery disease with emphasis on patient selection with particular attention to women, older persons, diabetic patients, and innovations in surgical techniques that may improve patient outcomes. PMID- 11061220 TI - Lipids--how low do you go: plaque regression and passivation. AB - Cholesterol-lowering and its relationship to acute coronary syndromes is the focus of this article. The pathophysiology of plaque formation and plaque rupture are briefly discussed as is the current thinking on the impact of cholesterol lowering on these processes. How cholesterol affects endothelial function and plaque passivation (the complex process of stabilizing the active plaque at risk of rupture) are key elements. A few hallmark clinical trials and their impact on coronary heart disease (CHD) clinical event rates are discussed as is the impact of statin therapy on CHD management. A brief description of the National Cholesterol Education Program and its recommendations for primary and secondary prevention are included. PMID- 11061221 TI - Pharmacology: antiplatelet and antithrombin therapy in acute coronary syndromes. AB - Coronary artery disease is the leading cause of death in the United States. According to estimates, 1.25 million acute myocardial infarctions (AMI) occur in the United States each year, 500,000 of which result in death (half of those resulting from sudden death within the first hour). Worldwide, the annual death toll from AMI is approximately 4 million. Inhibitors of platelet function have been associated with decreased morbidity in patients with atherosclerotic cardiovascular disease. Reperfusion of occluded coronary arteries via thrombolytic therapy has markedly reduced mortality and morbidity resulting from events of acute myocardial infarction. This article addresses several antiplatelet and antithrombin agents currently in clinical use throughout the United States. PMID- 11061222 TI - Factors influencing blood pressure: development of a risk model. AB - A review of the literature suggests that blood pressure (BP) is multifactorial and is affected by the interactions of genetics, physiology, responses to the environment, and lifestyle factors that have increasing influence as one ages. The effect of these factors on hypertension (HTN) risk as one ages is depicted in the author's Hypertension Risk Model. The model emphasizes HTN risk in the older adult--age 50 and older--and shows the interaction of the factors influencing HTN development and BP assessment methods. Implications of three methods of BP assessment--(1) static BP, (2) 24-hour ambulatory blood pressure monitoring, and (3) clinic blood pressure reactivity (BPR) protocol--are discussed. The model may be useful for understanding factors that contribute to HTN and for guiding BP assessment for clinical researchers. Addressing the factors associated with the sympathetic system activity from various environmental stressors requires assessing dynamic BP, particularly in the older hypertensive adult who has increased BPR. Further research should focus on ambulatory blood pressure studies in older adults that would provide the methods and instrumentation needed to assess HTN and therefore decrease mortality and morbidity in this population. PMID- 11061223 TI - On the similarity of properties in solution or in the crystalline state: a molecular dynamics study of hen lysozyme. AB - As protein crystals generally possess a high water content, it is assumed that the behaviour of a protein in solution and in crystal environment is very similar. This assumption can be investigated by molecular dynamics (MD) simulation of proteins in the different environments. Two 2ns simulations of hen egg white lysozyme (HEWL) in crystal and solution environment are compared to one another and to experimental data derived from both X-ray and NMR experiments, such as crystallographic B-factors, NOE atom-atom distance bounds, 3J(H N alpha) coupling constants, and 1H-15N bond vector order parameters. Both MD simulations give very similar results. The crystal simulation reproduces X-ray and NMR data slightly better than the solution simulation. PMID- 11061224 TI - Heteronuclear relayed E.COSY revisited: determination of 3J(H(alpha),C(gamma)) couplings in Asx and aromatic residues in proteins. AB - Constant-time 3D heteronuclear relayed E.COSY [Schmidt et al. (1996) J. Biomol. NMR, 7, 142-152], as based on generic 2D small-flip-angle HMQC-COSY [Schmidt et al. (1995) J. Biomol. NMR, 6, 95-105], has been modified to allow for quantitative determination of heteronuclear three-bond 3J(H(alpha),C(gamma)) couplings. The method is applicable to amino acid spin topologies with carbons in the gamma position which lack attached protons, i.e. to asparagine, aspartate, and aromatic residues in uniformly 13C-enriched proteins. The pulse sequence critically exploits heteronuclear triple-quantum coherence (HTQC) of CH2 moieties involving geminal H(beta) proton pairs, taking advantage of improved multiple quantum relaxation properties, at the same time avoiding scalar couplings between those spins involved in multiple-quantum coherence, thus yielding E.COSY-type multiplets with a splitting structure that is simpler than with the original scheme. Numerical least-squares 2D line-shape simulation is used to extract 3J(H(alpha),C(gamma)) coupling constants which are of relevance to side-chain chi1 dihedral-angle conformations in polypeptides. Methods are demonstrated with recombinant 15N,13C-enriched ribonuclease T1 and Desulfovibrio vulgaris flavodoxin with bound oxidized FMN. PMID- 11061225 TI - Direct measurement of 1H-1H dipolar couplings in proteins: a complement to traditional NOE measurements. AB - An intensity-based constant-time COSY (CT-COSY) method is described for measuring 1H-1H residual dipolar couplings of proteins in weakly aligned media. For small proteins, the overall sensitivity of this experiment is comparable to the NOESY experiment. In cases where the 1H-1H distances are defined by secondary structure, such as 1H(alpha)-1H(N) and 1H(N)-1H(N) sequential distances in alpha helices and beta-sheets, these measurements provide useful orientational constraints for protein structure determination. This experiment can also be used to provide distance information similar to that obtained from NOE connectivities once the angular dependence is removed. Because the measurements are direct and non-coherent processes, such as spin diffusion, do not enter, the measurements can be more reliable. The 1/r3 distance dependence of directly observed dipolar couplings, as compared with the 1/r6 distance dependence of NOEs, also can provide longer range distance information at favorable angles. A simple 3D, 15N resolved version of the pulse sequence extends the method to provide the improved resolution required for application to larger biomolecules. PMID- 11061226 TI - Differential multiple-quantum relaxation arising from cross-correlated time modulation of isotropic chemical shifts. AB - In this paper it is demonstrated that cross-correlated time modulation of isotropic chemical shifts ('conformational exchange') leads to differential relaxation of double- and zero-quantum coherences, respectively. Quantitative information can be obtained from the time dependence of the interconversion between the two two-spin coherences 2IxSx and 2IySy, induced by the differential relaxation. The effect is illustrated with an application to 13C,15N-labeled quail CRP2(LIM2), by studying 15N-1H(N) multiple-quantum relaxation. Significant cross-correlated fluctuations of isotropic chemical shifts were observed for residues which are part of a disordered loop region connecting two beta-strands in CRP2(LIM2). Differential 1H(N) and 15N exchange contributions to multiple quantum relaxation observed at these sites illustrate the complex interplay between hydrogen bonding events and conformational reorientations in proteins. PMID- 11061227 TI - Random coil chemical shifts in acidic 8 M urea: implementation of random coil shift data in NMRView. AB - Studies of proteins unfolded in acid or chemical denaturant can help in unraveling events during the earliest phases of protein folding. In order for meaningful comparisons to be made of residual structure in unfolded states, it is necessary to use random coil chemical shifts that are valid for the experimental system under study. We present a set of random coil chemical shifts obtained for model peptides under experimental conditions used in studies of denatured proteins. This new set, together with previously published data sets, has been incorporated into a software interface for NMRView, allowing selection of the random coil data set that fits the experimental conditions best. PMID- 11061228 TI - Simulations of NMR pulse sequences during equilibrium and non-equilibrium chemical exchange. AB - The McConnell equations combine the differential equations for a simple two-state chemical exchange process with the Bloch differential equations for a classical description of the behavior of nuclear spins in a magnetic field. This equation system provides a useful starting point for the analysis of slow, intermediate and fast chemical exchange studied using a variety of NMR experiments. The McConnell equations are in the mathematical form of an inhomogeneous system of first-order differential equations. Here we rewrite the McConnell equations in a homogeneous form in order to facilitate fast and simple numerical calculation of the solution to the equation system. The McConnell equations can only treat equilibrium chemical exchange. We therefore also present a homogeneous equation system that can handle both equilibrium and non-equilibrium chemical processes correctly, as long as the kinetics is of first-order. Finally, the same method of rewriting the inhomogeneous form of the McConnell equations into a homogeneous form is applied to a quantum mechanical treatment of a spin system in chemical exchange. In order to illustrate the homogeneous McConnell equations, we have simulated pulse sequences useful for measuring exchange rates in slow, intermediate and fast chemical exchange processes. A stopped-flow NMR experiment was simulated using the equations for non-equilibrium chemical exchange. The quantum mechanical treatment was tested by the simulation of a sensitivity enhanced 15N-HSQC with pulsed field gradients during slow chemical exchange and by the simulation of the transfer efficiency of a two-dimensional heteronuclear cross-polarization based experiment as a function of both chemical shift difference and exchange rate constants. PMID- 11061229 TI - Identification of compounds with binding affinity to proteins via magnetization transfer from bulk water. AB - A powerful screening by NMR methodology (WaterLOGSY), based on transfer of magnetization from bulk water, for the identification of compounds that interact with target biomolecules (proteins, RNA and DNA fragments) is described. The method exploits efficiently the large reservoir of H2O magnetization. The high sensitivity of the technique reduces the amount of biomolecule and ligands needed for the screening, which constitutes an important requirement for high throughput screening by NMR of large libraries of compounds. Application of the method to a compound mixture against the cyclin-dependent kinase 2 (cdk2) protein is presented. PMID- 11061230 TI - Assignment of the 1H, 13C, and 15N resonances of the DNA binding domain of gpNu1, a genome packaging protein from bacteriophage lambda. PMID- 11061231 TI - Sequence-specific 1H, 13C and 15N resonance assignments of the major cherry allergen Pru a 1. PMID- 11061232 TI - Sequence-specific resonance assignment of the Ras-binding domain of AF6. PMID- 11061233 TI - Backbone 1H, 13C, and 15N resonance assignments of an 18.2 kDa protein, E. coli peptidyl-prolyl cis-trans isomerase b (EPPIb). PMID- 11061234 TI - Psychiatric-mental health nursing on the Web. PMID- 11061235 TI - Assessment of inpatient treatment of persons with schizophrenia: implications for practice. AB - This descriptive study examined a stratified random sample by month of 457 persons with schizophrenia hospitalized on an inpatient unit at a nonprofit hospital located in Honolulu, HI in 1997. White patients were overrepresented in the sample when compared to the number in the general population. Part-Hawaiian patients had longer lengths of stay. Women were more likely to be admitted involuntarily than men. The majority of the patients were prescribed atypical antipsychotic medications. Only 8.8% of the patients were in restraints during their hospitalization. Problematic areas were patient teaching, monitoring of medication side effects, treatment planning, and discharge planning. The study site is fully accredited, attending physicians were all psychiatrists, and the majority of the nurses were certified. How pervasive there deficiencies are in other settings is not known. The findings of the study were used to revise a schizophrenic care path used on the inpatient unit. PMID- 11061236 TI - A view of prescriptive practice collaboration: perspectives of psychiatric-mental health clinical nurse specialists and psychiatrists. AB - A majority of states require collaborative prescribing agreements between advanced practice nurses and physicians. Unfortunately, there is limited research describing the collaboration that occurs between the clinicians who have such prescribing agreements. This exploratory study identifies the characteristics, activities, and outcomes of collaboration between psychiatric-mental health clinical nurse specialists (CNSs) with prescribing agreements and their collaborating psychiatrists. Surveys were sent to all the 73 prescribing psychiatric-mental health CNSs identified by the Minnesota Board of Nursing in 1998 and their primary collaborating psychiatrists. Forty-nine CNSs and 32 psychiatrists returned the surveys with 31 matched collaborating dyads identified. Overall satisfaction with the collaborative relationship was high, CNSs (chi = 4.34/5) and psychiatrists (chi = 4.46/5). Good communication, trust, shared goals for patient outcomes, shared professional values, and respect for clinical competency were identified as important characteristics for effective collaboration. CNSs identified increased professional growth and job satisfaction as professional benefits, while psychiatrists reported shared workload responsibilities. Fewer than half of the CNSs and psychiatrists perceived professional liability as a professional constraint. Psychiatric-mental health CNSs and psychiatrists agreed that the continuity of patient care and efficient access to mental health care were patient benefits. The statistically significant differences between the CNSs' and psychiatrists' responses were related to the number of years they had been in practice, the number of years the CNS had been a prescriber, and the length of time the CNS and psychiatrist had worked together within a collaborative prescribing agreement. PMID- 11061237 TI - Opening caregiver minds: National Alliance for the Mentally Ill's (NAMI) provider education program. AB - The belief that poor parenting and dysfunctional families give rise to mental illness has been perpetuated by psychodynamic and family systems theories that lack supporting scientific evidence, and interventions based on these theories have failed to produce clinical improvements. Nevertheless the National Alliance for the Mentally III (NAMI) found that many clinical training programs continue to teach these outdated theories and interventions and that the mental health system is often destructive to family systems. This article describes a new 10 week program that is designed to educate service providers that will include families in the care of their chronically ill loved one. The program is based on a competence and adaptation rather than a pathology foundation and it shifts the discourse from causes to effects of illness. PMID- 11061238 TI - Helping women caregivers obtain support: barriers and recommendations. AB - Women frequently assume the role of caregiver, and the demands of the caregiving role may lead to stress. Social support may moderate the effects of this stress. This article describes characteristics of effective social support, barriers to obtaining adequate support, and recommendations for how a woman in a caregiving role can obtain the support she needs. Nurses caring for women and families throughout the life span can use this information to help caregivers to obtain the necessary social support. PMID- 11061240 TI - ISPN 3-year strategic plan. International Society of Psychiatric Nurses. PMID- 11061239 TI - Frontotemporal dementia: a different kind of dementia. AB - Frontotemporal dementia (FTD) is an umbrella term for a number of uncommon illnesses, including Pick's disease, which affect the frontal and/or temporal lobes of the brain and produce a dementia syndrome that is quite characteristic but unfamiliar to most clinicians. It is the third most common cause of cortical dementia. An overview of clinical manifestations, diagnosis, and management of FTD is provided with some specific information in relation to Pick's disease and a case example. Caregivers, in particular, need the education and support that could be provided by informed nurses. PMID- 11061241 TI - Fas(t) track to apoptosis in MS: TNF receptors may suppress or potentiate CNS demyelination. PMID- 11061242 TI - What causes intracerebral hemorrhage during warfarin therapy? PMID- 11061243 TI - Assessment: prevention of post-lumbar puncture headaches: report of the therapeutics and technology assessment subcommittee of the american academy of neurology. PMID- 11061244 TI - Pharmacologic treatment of pain in polyneuropathy. AB - Tricyclic antidepressants and anticonvulsants have become the mainstay in the treatment of pain in polyneuropathy. Within the last decade, controlled trials have shown that numerous other drugs relieve such pain. To estimate the efficacy of the different treatments, the authors identified all placebo-controlled trials and calculated numbers needed to treat (NNT) to obtain one patient with more than 50% pain relief. The NNT was 2.6 for tricyclic antidepressants, 6.7 for selective serotonin reuptake inhibitors, 2.5 for anticonvulsant sodium channel blockers, 4.1 for the anticonvulsant calcium channel blocker gabapentin, and 3.4 for the mixed opioid and monoaminergic drug tramadol, as calculated from a sufficiently large number of patients. Favorable point estimates of NNT of 1.9 for the NMDA antagonist dextromethorphan and 3.4 for L-dopa were determined from a limited number of data. For capsaicin, the NNT calculated from many exposed patients was 5.9, but most of the data are controversial owing to trial methodology. Finally, the NNT for the antiarrhythmic sodium channel blocker mexiletine was 38, but this value may be biased because of a lack of dichotomous data in several positive trials. Tricyclic antidepressants are at the moment still the drugs of first choice, and drugs such as gabapentin, carbamazepine, and tramadol may be tried if contraindications or tolerability problems are encountered with the tricyclics. PMID- 11061245 TI - Defective T cell fas function in patients with multiple sclerosis. AB - BACKGROUND: Fas (CD95) triggers programmed cell death and is involved in shutting off the immune response. Inherited deleterious mutations hitting Fas or its signaling pathway cause autoimmune/lymphoproliferative syndrome (ALPS). OBJECTIVE: To assess the possibility that decreased Fas function plays a role in development of MS. METHODS: The authors evaluated Fas function in long-term T cell lines (21 days of culture) from 32 patients with relapsing-remitting MS (RRMS), 15 with secondary progressive MS (SPMS), and 15 with primary progressive MS (PPMS) by assessing cell survival upon Fas triggering by monoclonal antibodies (Mab). RESULTS: Fas-induced cell death was significantly lower in all patient groups than in controls, and lower in SPMS than in RRMS. Moreover, 8/15 patients with PPMS, 10/15 with SPMS, and 8/32 with RRMS were frankly resistant to Fas. Frequency of resistance to Fas-induced cell death was significantly higher in all patient groups than in controls (2/75), and higher in SPMS than in RRMS. The findings that the parents of two Fas-resistant patients were Fas-resistant and that fusion of T cells from two Fas-resistant patients with Fas-sensitive HUT78 cells gave rise to Fas-resistant hybrid lines suggest that Fas-resistance is due to inherited alterations of the Fas signaling pathway, with production of molecules exerting a dominant negative effect on a normal Fas system. CONCLUSIONS: Defects of the immune response shutting-off system may be involved in the pathogenesis of MS, particularly in its progressive evolution. PMID- 11061246 TI - Apoptosis mediators fasL and TRAIL are upregulated in peripheral blood mononuclear cells in MS. AB - OBJECTIVE: To investigate the expression of apoptosis-inducing ligand and receptor molecules in patients with MS. BACKGROUND: Dysregulation of apoptosis may induce autoimmune conditions, possibly through inadequate termination of immune responses, and could be of importance for pathogenesis of MS. METHODS: Messenger RNA (mRNA) levels of two apoptosis-related members of the tumor necrosis factor (TNF) receptor family, Fas and TNF-related apoptosis-inducing ligand (TRAIL) receptor 2 (TRAIL-R2), and their ligands, Fas ligand (FasL) and TRAIL, were quantified by competitive reverse transcription PCR in unstimulated peripheral blood mononuclear cells in 47 untreated patients with MS and 46 control subjects. RESULTS: The expression of FasL was increased in patients with MS compared with healthy control subjects. Analysis of clinical subgroups revealed that the increase was marked in relapsing-remitting MS, being especially high in remission (p = 0.0002), but less so in chronic progressive MS (p = 0.14). Compared with healthy control subjects, TRAIL mRNA levels were also upregulated in patients with MS (p = 0.0001) but did not differ between clinical subgroups. The expression of TRAIL-R2 was slightly elevated in patients with MS (p = 0.02) whereas the expression of Fas was similar in patients and control subjects. The ratio of expression levels for two isoforms of TRAIL-R2, TRICK2a and TRICK2b, in patients with MS differed from healthy control subjects (p = 0.04). CONCLUSIONS: There was increased expression of both FasL and TRAIL in peripheral blood lymphocytes. It remains to be determined whether this increased expression represents a disease-promoting autoimmune process or is merely the effect of a secondary compensatory mechanism that downregulates the inflammatory response. PMID- 11061247 TI - Fatigue and declines in cognitive functioning in multiple sclerosis. AB - OBJECTIVE: To determine whether cognitive fatigue, defined as a decline in cognitive performance over a single testing session, could be identified in MS. METHODS: Forty-five individuals with MS and 14 healthy control participants completed a 4-hour session of cognitive testing that involved a baseline neuropsychological battery, a continuous effortful cognitive task (completing mental arithmetic problems administered on a computer), and a repeat neuropsychological battery. Self-report measures of fatigue and affect were completed before each step of the testing session. RESULTS: The pattern of change in cognitive performances over the testing session significantly differed between the MS and control participants. Individuals with MS showed declines on measures of verbal memory and conceptual planning, whereas the control participants showed improvement. Although there were no significant differences between the groups on any of the baseline cognitive measures, the MS participants performed worse than the control subjects on tests of visual memory, verbal memory, and verbal fluency that were repeated following the continuous effortful cognitive task. Both MS and control participants reported increased mental and physical fatigue across the testing session compared with their baseline values. CONCLUSION: Individuals with MS show declines in cognitive performance during a single testing session and fail to show the improvement exemplified by healthy control subjects. PMID- 11061248 TI - Magnetization transfer imaging to monitor the evolution of MS: a 1-year follow-up study. AB - OBJECTIVES: To assess the sensitivities of magnetization transfer imaging (MTI) derived measures in detecting changes over time of macro- and microscopic lesion burdens in different MS phenotypes and to compare them with those of T2-weighted and T1-weighted lesion volumes. METHODS: A total of 96 patients were studied: 39 with relapsing-remitting MS (RRMS), 19 with secondary progressive MS (SPMS), nine with primary progressive MS, and nine with benign MS; 20 with clinically isolated syndromes suggestive of MS at presentation; and 20 healthy subjects. Brain T2 weighted, T1-weighted, and MTI scans were obtained at baseline and after 12 months. The authors measured T2-weighted and T1-weighted lesion volumes and average lesion MT ratio (MTR). The authors also derived MTR histograms from whole brain tissue (WBT) and normal-appearing brain tissue (NABT). RESULTS: In healthy control subjects, there was no significant change of any of the MTR histogram parameters. At follow-up, in the entire patient group, T2-weighted lesion volume significantly increased and average lesion MTR, WBT-MTR, NABT-MTR, and histogram peak positions significantly decreased. Patients with RRMS and SPMS had significantly higher changes in T2-weighted lesion volume and all the MTI-derived metrics compared with the other subgroups. MTI changes were more prominent (and significantly different) in patients with SPMS than in those with RRMS. Compared with patients with benign MS, patients with RRMS had significantly greater changes in T2-weighted lesion volume and WBT- and NABT-MTR metrics. Compared with patients with SPMS, patients with primary progressive MS had similar changes of T1-weighted and T2-weighted lesion volumes, but significantly lower changes of MTI-derived measures. CONCLUSIONS: MTI-derived measures are sensitive for detecting MS-related changes and might provide valuable outcome measures when assessing treatment effects in clinical trials of patients with MS. PMID- 11061249 TI - Warfarin-associated hemorrhage and cerebral amyloid angiopathy: a genetic and pathologic study. AB - BACKGROUND: Intracerebral hemorrhage (ICH) is the most feared complication of warfarin therapy. The pathogenesis of this often-fatal complication remains obscure. Cerebral amyloid angiopathy (CAA) is a major cause of spontaneous lobar hemorrhage in the elderly and is associated with specific alleles of the APOE gene. OBJECTIVE: To assess the role of CAA in warfarin-associated ICH. METHODS: Clinical characteristics and APOE genotype were compared between 41 patients with warfarin-related ICH (from a cohort of 59 consecutive patients aged > or = 65 years with supratentorial ICH on warfarin) and 66 randomly selected individuals aged > or = 65 years without ICH taking warfarin. In addition, all neuropathologic specimens from ICH patients were reviewed for the presence and severity of CAA. RESULTS: Hemorrhages tended to be in the lobar regions of the brain, and most (76%) occurred with an international normalized ratio of < or = 3.0. The APOE epsilon2 allele was overrepresented among patients with warfarin associated lobar hemorrhage (allele frequency 0.13 versus 0.04 in control subjects; p = 0.031). After controlling for other variables associated with ICH, carriers of the epsilon2 allele had an OR of 3.8 (95% CI, 1.0 to 14.6) for lobar ICH. CAA was pathologically diagnosed as the cause of lobar hemorrhage in 7 of 11 patients with available tissue samples. CONCLUSIONS: CAA is an important cause of warfarin-associated lobar ICH in the elderly. Although diagnosis of CAA before hemorrhage is not yet possible, these data offer hope that future patients at high risk for hemorrhage may be identified before initiation of warfarin therapy. PMID- 11061250 TI - Predicting prognosis after stroke: a placebo group analysis from the National Institute of Neurological Disorders and Stroke rt-PA Stroke Trial. AB - BACKGROUND: Physicians are often asked to predict outcome after acute stroke. Very little information is available that can reliably predict the likelihood of severe disability or death. OBJECTIVE: To develop a practical method for predicting a poor outcome after acute ischemic stroke. METHODS: Data from the placebo arms of Parts 1 and 2 of the National Institute of Neurological Disorders and Stroke rt-PA [recombinant tissue plasminogen activator] Stroke Trial were used to identify variables that could predict a poor outcome, defined as moderately severe disability, severe disability, or death (Modified Rankin Scale score >3) 3 months after stroke. RESULTS: Baseline variables that predicted poor outcome were the NIH Stroke Scale (NIHSS) >17 plus atrial fibrillation, yielding a positive predictive value (PPV) of 96% (95% CI, 88 to 100%). The best predictor at 24 hours was NIHSS >22, yielding a PPV of 98% (95% CI, 93 to 100%). The best predictor at 7 to 10 days was NIHSS >16, yielding a PPV of 92% (95% CI, 85 to 99%). CONCLUSIONS: Patients with a severe neurologic deficit after acute ischemic stroke, as measured by the NIHSS, have a poor prognosis. During the first week after acute ischemic stroke, it is possible to identify a subset of patients who are highly likely to have a poor outcome. These findings require confirmation in a separate study. PMID- 11061251 TI - Pulmonary arteriovenous malformations: cerebral ischemia and neurologic manifestations. AB - BACKGROUND: There is an increasingly recognized association between pulmonary arteriovenous malformations (PAVM) and cerebral ischemia, frequently attributed to paradoxical embolization. PAVM occur in 20 to 30% of the hereditary hemorrhagic telangiectasia (HHT) population. OBJECTIVE: To evaluate the risk determinants for cerebral ischemia and neurologic manifestations in patients with PAVM. METHODS: A retrospective cross-sectional study was performed on consecutive patients admitted between 1988 and 1992 for treatment of PAVM. The number of PAVM, feeding artery (FA) diameters, and aneurysmal sizes were determined by pulmonary angiography. Patients were categorized as having single or multiple PAVM with an FA diameter of > or = 3 mm. History, examination, and cerebral imaging studies were used to determine the prevalence of neurologic manifestations. Patients were defined as having cerebral paradoxical embolization if there was radiologic evidence of cortical infarction. RESULTS: There were 75 cases: 26 single PAVM and 49 multiple PAVM. Cortical infarction was present in 14% of patients with single PAVM. Patients with multiple PAVM had a greater prevalence of any infarction (OR 3.2; 95% CI, 1.2 to 9.44, p = 0.030), cortical infarctions (OR 2.3; 95% CI, 0.58 to 9.2, p = 0.230), subcortical infarctions (OR 2.1; 95% CI, 0.58 to 7.95, p = 0.249), abscesses (OR 2.3; 95% CI, 0.46 to 11.94; p = 0.295), and seizures (OR 6.4, 95% CI 0.77 to 53.2, p = 0.054). Patients with multiple PAVM had markedly greater odds of having any clinical or radiologic evidence of cerebral ischemic involvement (OR 4.5; 95% CI, 1.47 to 14; p = 0.008). CONCLUSION: There is a strong association between single PAVM and various neurologic manifestations. The prevalence is greater for patients with multiple PAVM, suggesting increased predisposition for paradoxical embolization with a greater number of malformations. PMID- 11061252 TI - A randomized, controlled trial of high-dose dextromethorphan in facial neuralgias. AB - BACKGROUND: NMDA glutamate receptor antagonists such as ketamine and dextromethorphan reduce pain in certain neuropathic pain conditions. However, there have been no controlled trials of NMDA antagonists in facial neuralgias. METHODS: A randomized, double-blind, crossover trial compared 6 weeks of oral dextromethorphan with active placebo (low-dose lorazepam) in 19 patients, stratified into three groups: 11 with facial pain and possible trigeminal neuropathy, five with anesthesia dolorosa, and three with idiopathic trigeminal neuralgia. Dosage was titrated in each patient to the highest level reached without disrupting normal activities. RESULTS: Patients completing the trial included 10 with possible trigeminal neuropathy, four with anesthesia dolorosa, and two with trigeminal neuralgia. In patients with possible trigeminal neuropathy and anesthesia dolorosa, dextromethorphan decreased pain by a mean of only 2 to 4%, and these estimates were not significant. Both patients with trigeminal neuralgia had more pain during dextromethorphan treatment than during placebo treatment. Of three patients who demonstrated an analgesic response to dextromethorphan during the main trial, only one repeatedly responded in four subsequent confirmatory drug-placebo crossovers. CONCLUSIONS: Dextromethorphan shows little or no analgesic efficacy in pain due to possible trigeminal neuropathy and anesthesia dolorosa. Additional trials are necessary to conclusively evaluate the efficacy of NMDA-receptor antagonists in trigeminal neuralgia. PMID- 11061253 TI - Lack of efficacy of riluzole in the treatment of peripheral neuropathic pain conditions. AB - OBJECTIVE: To assess the efficacy, tolerability, and safety of riluzole in the treatment of peripheral neuropathic pain conditions. BACKGROUND: Both basic and clinical research has demonstrated that drugs with sodium channel and NMDA antagonism can be effective in alleviating neuropathic pain. Riluzole, a drug currently used for treatment of ALS, possesses these properties. It was hypothesized that riluzole would be effective in reducing the pain in subjects with peripheral neuropathic pain. METHODS: Two randomized, placebo-controlled, crossover studies were performed at two sites. Study 1 compared 100 mg/day of riluzole (the currently recommended dosage for treatment of ALS) versus placebo, and Study 2 compared 200 mg/day of riluzole versus placebo. Each treatment phase (both studies) was 2 weeks long, separated by 2-week wash-out periods. Outcome measures included change in the score on a 100-mm pain intensity visual analog scale, the Neuropathic Pain Scale, allodynia, hyperalgesia, and preference for study treatment phase. RESULTS: Twenty-two subjects completed Study 1, and 21 subjects completed Study 2. Four subjects (two from each study) discontinued the study because of intolerable side effects. No statistical difference was found for any study outcome measure between riluzole and placebo for either study. In Study 1, pain intensity was more likely to increase than decrease with riluzole (mean treatment difference 8.7 mm; 95% CI -19.5 to +2.1 mm). In Study 2, very slight pain reduction was observed with riluzole compared with placebo (mean treatment difference 1.4 mm; 95% CI -5.1 to +8.0 mm). In both studies, the majority of subjects chose "no change" in pain on the category relief scale after placebo and riluzole treatment phases. On study completion, no treatment preference was reported by 76% of the subjects in Study 1 and by 61% of the subjects in Study 2. CONCLUSIONS: Doses of riluzole at (100 mg) or above (200 mg) those used for the treatment of ALS were not effective in alleviating peripheral neuropathic pain. PMID- 11061254 TI - Changes in spinal cord excitability in patients affected by ulnar neuropathy. AB - OBJECTIVE: To evaluate whether ulnar neuropathy could induce changes in spinal cord and motor cortex excitability and therefore predispose to development of focal dystonia. BACKGROUND: A high incidence of ulnar neuropathy has been observed in patients with musician's cramp. Polygraphic electromyograph recordings in patients with entrapment of the ulnar nerve at the elbow have demonstrated long-duration bursts of co-contraction in antagonistic muscles, similar to those observed in focal dystonia. METHODS: All control subjects and 12 patients with ulnar neuropathy underwent an electrophysiologic protocol consisting of polygraphic recordings of a repetitive tapping task of the fourth finger, assessment of reciprocal inhibition in forearm muscles, and investigation of motor cortex excitability after paired transcranial magnetic stimulation. RESULTS: Eleven of 12 patients with ulnar neuropathy showed a loss of alternation and of well-formed bursts in both flexor and extensor muscles. Evaluation of reciprocal inhibition in these patients revealed a reduction in the amount of inhibition in the disynaptic and presynaptic phases. None of the patients presented with a clinically evident dystonia of the upper limb. The study of intracortical excitability after paired shocks did not reveal any difference in the amount of intracortical inhibition and facilitation compared with the control group. CONCLUSIONS: A peripheral nerve injury can induce a rearrangement of reciprocal inhibition circuits at the spinal cord level. These changes might predispose to the development of a focal dystonia. However, it is likely that another, yet unknown, factor is required to alter the intracortical circuits and produce a clinically evident dystonia. PMID- 11061255 TI - The influence of normal aging on the cortical processing of a simple motor task. AB - OBJECTIVE: To assess the influence of normal aging on the cortical physiology of motor behavior. METHODS: The authors studied cortical activation in eight elderly (55 to 76 years of age) and eight younger (18 to 27 years of age) healthy subjects while they performed a simple motor task. A 28-channel EEG was recorded; task-related power changes associated with repetitive, metronome-paced (1 Hz) finger movements were computed as a measure of cortical activation. RESULTS: Distinct, age-dependent activation patterns were expressed in four distinct frequency ranges: low-alpha (10 to 11 Hz), high-alpha (12 to 13 Hz), low-beta (16 to 17 Hz), and high-beta (22 to 23 Hz) bands. The main findings were a greater overall activation and, more specifically, a pronounced bilateral activation of sensorimotor regions in elderly subjects for both alpha bands. Additionally, in the elderly subjects there was increased activation of the mesial frontocentral cortex (supplementary motor area region) in the high-beta band, whereas younger volunteers had a prominent activation of the left lateral premotor and sensorimotor region in this frequency range. CONCLUSIONS: These findings demonstrate that the functional anatomy of the human motor system changes during normal aging. It appears that, for a given motor task, the aging brain recruits additional primary sensorimotor and premotor regions of both hemispheres. PMID- 11061256 TI - A novel de novo mutation in the desmin gene causes desmin myopathy with toxic aggregates. AB - OBJECTIVE: To determine the cause and pathogenic mechanisms of a 21-year-old patient's cardioskeletal myopathy. The patient's muscle atrophy and weakness began in distal parts of limbs; cardiac and facial muscles were later involved. BACKGROUND: Desmin myopathy is a skeletal myopathy often associated with cardiomyopathy, caused by mutations in the desmin gene and characterized by desmin accumulation in affected muscle fibers, a leading marker of myofibrillar myopathies. Two kinds of deletions and seven missense mutations in the desmin gene have been identified. METHODS: Clinical examination, electron microscopy of muscle tissue, two-dimensional gel electrophoresis, DNA sequencing, restriction enzyme analysis, and gene transfection were performed. RESULTS: Electron microscopy showed disruption of sarcomeres at Z discs and electron-dense aggregates in biopsied skeletal and heart muscle. Two-dimensional gel electrophoresis of the patient's skeletal muscle proteins showed massive accumulation of desmin. The authors identified a novel desmin mutation, L385P in one allele in the carboxyl end of the rod domain 2B in the patient's leukocytes and skeletal muscle; neither parent had the mutation. Serologic study and DNA markers confirmed the de novo mutation. A peptide harboring desmin rod domains 2A and 2B with L385P tagged with green fluorescent protein induced cytoplasmic aggregates, nuclear DNA condensation, and cell death. CONCLUSIONS: A novel de novo mutation, L385P, causes desmin myopathy. An expression study indicated the toxic effect of the L385P mutation. PMID- 11061257 TI - Rapid-onset dystonia-parkinsonism: a clinical and genetic analysis of a new kindred. AB - BACKGROUND: Rapid-onset dystonia-parkinsonism (RDP) is an autosomal dominant disorder linked to chromosome 19q13 that is characterized by sudden onset of primarily bulbar and upper limb dystonia with parkinsonism. METHODS: The authors evaluated 12 individuals from three generations of an Irish family and obtained detailed medical records on a deceased member. The authors describe the clinical, psychiatric, and genetic features of the affected individuals. RESULTS: Five of eight affected members developed sudden-onset (several hours to days) dystonia with postural instability. Four of the five also had bulbar symptoms. Two have stable focal or segmental limb dystonia. One has intermittent hemidystonia with dysarthria that comes on abruptly in times of stress or anxiety. Three had a history of profound difficulty socializing, and at presentation two developed depression. Three patients had a trial of dopamine agonists without benefit. Genetic analysis suggests linkage to chromosome 19 with lod score of 2.1 at zero recombination. CONCLUSION: This is the third reported family with chromosome 19q13 rapid-onset dystonia-parkinsonism. Psychiatric morbidity appeared common in affected members of this family and may be part of the RDP phenotype. PMID- 11061258 TI - Motor imagery in normal subjects and in asymmetrical Parkinson's disease: a PET study. AB - OBJECTIVE: To investigate, using PET and H2(15)O, brain activation abnormalities of patients with PD during motor imagery. To determine whether motor imagery activation patterns depend on the hand used to complete the task. BACKGROUND: Previous work in PD has shown that bradykinesia is associated with slowness of motor imagery. METHODS: The PET study was performed in eight patients with PD with predominantly right-sided akinesia, and in eight age-matched control subjects, all right-handed. Regional cerebral blood flow was measured by PET and H2(15)O while subjects imagined a predetermined unimanual externally cued sequential movement with a joystick with either the left or the right hand, and during a rest condition. RESULTS: In normal subjects, the prefrontal cortex, supplementary motor area (SMA), superior parietal lobe, inferior frontal gyrus, and cerebellum were activated during motor imagery with either the left or the right hand. Contralateral primary motor cortex activation was noted only when the task was imagined with the right (dominant) hand, whereas activation of the dorsolateral prefrontal cortex was observed only during imagery with the left hand. In patients with PD, motor imagery with the right ("akinetic") hand was characterized by lack of activation of the contralateral primary sensorimotor cortex and the cerebellum, persistent activation of the SMA, and bilateral activation of the superior parietal cortex. Motor imagery with the left ("non akinetic") hand was also abnormal, with lack of activation of the SMA compared with controls. CONCLUSIONS: In patients with PD with predominantly right-sided akinesia, brain activation during motor imagery is abnormal and may appear even with the less affected hand. In normal subjects, brain activation during motor imagery depends on the hand used in the imagined movement. PMID- 11061259 TI - Obstructive sleep apnea is common in medically refractory epilepsy patients. AB - BACKGROUND: Previous reports have documented the coexistence of obstructive sleep apnea (OSA) and epilepsy and the therapeutic effects of treatment on seizure frequency and daytime sleepiness. The authors' objective was to determine the prevalence of OSA and its association with survey items in a group of patients with medically refractory epilepsy undergoing polysomnography (PSG). METHODS: Thirty-nine candidates for epilepsy surgery without a history of OSA underwent PSG as part of a research protocol examining the relationship of interictal epileptiform discharges to sleep state. Subjects also completed questionnaires about their sleep, including validated measures of sleep-related breathing disorders (Sleep Apnea Scale of the Sleep Disorders Questionnaire [SA/SDQ]) and subjective daytime sleepiness (Epworth Sleepiness Scale [ESS]). RESULTS: One third of subjects had OSA, defined by a respiratory disturbance index (RDI) > or = 5. Five subjects (13%) had moderate to severe OSA (RDI > 20). Subjects with OSA were more likely to be older, male, have a higher SA/SDQ score, and more likely to have seizures during sleep than those without OSA (p < 0.05). Seizure frequency per month, the number or type of antiepileptic drugs (AED) prescribed, the localization of seizures (temporal versus extratemporal), and the ESS were not statistically different between the two groups. CONCLUSIONS: In our sample, previously undiagnosed obstructive sleep apnea was common, especially among men, older subjects, and those with seizures during sleep. The impact of treating OSA on seizure frequency and daytime sleepiness in medically refractory epilepsy patients warrants further controlled study. PMID- 11061260 TI - Caregivers' preferences for the treatment of patients with Alzheimer's disease. AB - OBJECTIVES: To learn caregivers' preferences for the treatment of AD with a disease-slowing therapy, and to identify relationships between these preferences and the characteristics of caregivers and patients. METHODS: A structured interview with 40 caregivers of patients with AD. Preferences were measured for an AD-slowing medicine with the benefits of a gain in survival and a delay to nursing home placement (NHP), and risks of three degrees of severity of GI bleeding. RESULTS: Using a six-point scale ranging from "not at all important" to "extremely important," the median rating of the importance of survival as a treatment benefit was "very important" and of a delay to NHP was "extremely important." Fifty-five percent of the caregivers identified a benefit more important than these two benefits. Qualitative data showed that caregivers' reasons for these preferences featured the importance of patient quality of life, and the preservation of patient cognition and function. Bivariate analyses showed that increasing importance of the benefit of survival was related to higher assessments of the patient's health and quality of life, and lower ratings of the caregiver's experience of burden. In order to slow disease progression by 1 year, 25/38 (66%) of caregivers would accept some risk of death from gastrointestinal bleeding. Regression models showed that risk tolerance was higher among caregivers who were working, adult children caring for early stage patients or from families with an history of dementia. CONCLUSIONS: Caregivers generally are willing to tolerate notable amounts of risk to slow AD progression. Factors that describe the caregiver's experience and perception of the patient and the patient caregiver relationship influence how they want to treat the disease. This information may be useful for decisions about how to study and prescribe AD treatments. PMID- 11061261 TI - The Goltz-Ferrier debates and the triumph of cerebral localizationalist theory. AB - OBJECTIVE: To analyze the significance of the Goltz-Ferrier debates held at the International Medical Congress of 1881 for the development of ideas on cerebral localization. BACKGROUND: Cerebral localization was the subject of vigorous debate throughout the 19th century. At the Congress of 1881, David Ferrier, a leading proponent of cerebral localization, and Friedrich Leopold Goltz, an equally prominent anti-localizationist, had the opportunity to present their experimental research before 3,000 of the world's leading medical figures. METHODS: The authors reviewed and translated the presentations by Goltz and Ferrier at the Congress and supporting publications in contemporary medical journals. RESULTS: In his presentation to the Physiology Section, Goltz criticized localizationists for their widely divergent conclusions about the exact anatomic sites of cortical centers and for their failure to adequately explain functional restitution after cortical ablations. He noted that localizationist theories could, like an apple, "look very tempting and still have a worm inside." He described his own studies on massive decerebrations in dogs and noted that despite complete destruction of the cortices of both hemispheres these animals failed to exhibit motor weakness or deficits in primary sensation. Ferrier noted that Goltz's results were irreconcilable with his own experiments in monkeys, in which circumscribed lesions produced clear and reproducible functional deficits. Both investigators exhibited animals with cortical ablations. Ferrier's presentation of a hemiplegic monkey prompted Charcot's famous utterance, "C'est un malade!" ["It's a patient!"]. A distinguished committee examined the brains of the animals, and confirmed that Ferrier had indeed succeeded in producing a circumscribed lesion in the frontoparietal cortex, whereas the cortical ablations in Goltz's dogs were much less widespread than anticipated. CONCLUSIONS: Ferrier's dramatic demonstration of the effects produced by localized lesions in macaques triumphed over Goltz's unitary view of brain function, providing a major impetus for the subsequent successful development of neurologic surgery. PMID- 11061262 TI - Repetitive transcranial magnetic stimulation does not replicate the Wada test. AB - The authors compared inferior frontal speech arrest from repetitive transcranial magnetic stimulation (rTMS) with bilateral Wada tests in 17 epilepsy surgery candidates. Although rTMS lateralization correlated with the Wada test in most subjects, rTMS also favored the right hemisphere at a rate significantly greater than the Wada test. Postoperative language deficits were more consistent with Wada results. Available methods for inducing speech arrest with rTMS do not replicate the results of Wada tests. PMID- 11061263 TI - TLE patients with postictal psychosis: mesial dysplasia and anterior hippocampal preservation. AB - The authors studied six patients with refractory temporal lobe epilepsy and postictal psychosis using quantitative MRI and histopathology, and compared the results with 45 patients with temporal lobe epilepsy without postictal psychosis. Total hippocampal volumes were not different between the two groups. However, patients with postictal psychosis had a relatively preserved anterior hippocampus, and temporal lobe dysplasia was more frequent (p = 0.006, chi-square test). These findings may be associated with the clinical symptoms. PMID- 11061264 TI - Adult myoclonic epilepsy: a distinct syndrome of idiopathic generalized epilepsy. AB - The authors present 11 cases of idiopathic generalized epilepsy that began in adulthood at a mean age of 39 years. All patients had myoclonic jerks, five had absence seizures, and nine had infrequent generalized tonic-clonic seizures. A majority had a family history of seizures. EEG in all patients showed generalized epileptiform abnormalities, whereas neuroimaging and neurologic examination results were normal. This series appears to represent a previously undescribed idiopathic generalized epilepsy syndrome of adult myoclonic epilepsy. PMID- 11061265 TI - Corpus callosum size in autism. AB - The size of the seven subregions of the corpus callosum was measured on MRI scans from 22 non-mentally retarded autistic subjects and 22 individually matched controls. Areas of the anterior subregions were smaller in the autistic group. In a subsample, measurements were adjusted for intracranial, total brain, and white matter volumes and the differences between groups remained significant. No differences were found in the other subregions. This observation is consistent with the frontal lobe dysfunction reported in autism. PMID- 11061266 TI - Adult-onset MLD: a gene mutation with isolated polyneuropathy. AB - A 22-year-old man presented with recurrent ulnar mononeuropathies and diffusely slow nerve conduction velocities. Arylsulfatase A (ASA) activity from leukocytes and fibroblasts was reduced, and urinary sulfatides were increased. Sural nerve biopsy revealed a reduction in myelinated fibers and Schwann cell inclusions. Results of studies of CNS integrity, including cranial MRI, evoked potentials, and neuropsychologic tests, were normal. Molecular genetic analyses revealed a novel homozygous missense mutation (Thr286Pro) in the ASA gene. PMID- 11061267 TI - CACNA1A gene de novo mutation causing hemiplegic migraine, coma, and cerebellar atrophy. AB - Familial hemiplegic migraine is caused by CACNA1A missense mutations in 50% of families, including all families with cerebellar ataxia. A patient with healthy parents, who experienced prolonged attacks of migraine with hemiplegia, coma, and seizures, is reported. The patient also had mental retardation, permanent cerebellar ataxia with cerebellar atrophy, and right-sided brain atrophy. This patient carried a de novo Tyr 1385 Cys mutation in the CACNA1A gene and illustrates a novel phenotype associated with CACNA1A mutations. PMID- 11061268 TI - Indomethacin reduces CSF pressure in intracranial hypertension. AB - The CSF-pressure-lowering effects of indomethacin in seven patients with idiopathic intracranial hypertension and one patient with symptomatic intracranial hypertension due to a non-space-occupying meningioma are reported. CSF opening pressure between 350 and 500 mm H2O (mean 400 mm H2O) was promptly reduced by 80 to 200 mm H2O (mean reduction, 139 mm H2O) for at least 10 minutes in all patients after IV administration of 50 mg indomethacin. Four patients had mild and transient side effects (dizziness). Indomethacin might be an alternative drug for treatment of intracranial hypertension. PMID- 11061269 TI - Complications during apnea testing in the determination of brain death: predisposing factors. AB - Apnea testing in brain death determination may result in cardiovascular complications. Hypotension occurred in 24% and cardiac arrhythmias occurred in <1% of the 145 apneic oxygenation procedures. Complications were noted in only 15% of apnea tests performed without any predisposing factors. Significantly more complications (39%) were observed in apnea tests with inadequate precautions, particularly in apnea tests without adequate preoxygenation (50%). PMID- 11061270 TI - Cortical MRI findings associated with rapid correction of hyponatremia. AB - The authors describe two patients with clinical manifestations of the osmotic demyelination syndrome (ODS) and unusual MRI findings of gadolinium-enhancing peripheral cortical abnormalities. They propose that these represent extrapontine manifestations of ODS because neither patient had a notable hypoxic-ischemic insult. Recognizing this imaging appearance is important because prognosis in ODS may be less uniformly grim than for hypoxia-ischemia. PMID- 11061271 TI - Thalamic microglial activation in ischemic stroke detected in vivo by PET and [11C]PK1195. AB - Using quantitative PET, the authors studied the binding of [11C]PK11195, a marker of activated microglia, in the thalamus of patients with chronic middle cerebral artery infarcts. All patients showed increased [11C]PK11195 binding in the ipsilateral thalamus, indicating the activation of microglia in degenerating projection areas remote from the primary lesion. A persistent increase in [11C]PK11195 binding suggests active, long-term thalamic microstructural changes after corticothalamic connection damage. PMID- 11061272 TI - A new mutation in the prion protein gene: a patient with dementia and white matter changes. AB - The authors describe the clinical characteristics, MRI abnormalities, and molecular findings in a patient with a novel variant of a two-octarepeat insertion mutation in the prion protein gene. This patient presented with moderately progressive dementia of presenile onset and gait ataxia. MRI showed extensive cortical atrophy and white matter abnormalities. The mutation consists of a two-octarepeat insertion mutation and irregularities in the nucleotide sequence of the octarepeat region. PMID- 11061273 TI - MRI findings in Mobius syndrome: correlation with clinical features. AB - The authors studied the MRI findings of three patients with Mobius syndrome. Mobius syndrome is a rare congenital disorder characterized by complete or partial facial diplegia accompanied by other cranial nerve palsies. MRI demonstrated brainstem hypoplasia with straightening of the fourth ventricle floor, indicating an absence of the facial colliculus. These MRI features suggest the diagnosis of Mobius syndrome and correlate with the clinical and neurophysiologic findings. PMID- 11061274 TI - MRI evidence of mesial temporal sclerosis in patients with psychogenic nonepileptic seizures. PMID- 11061275 TI - Prevalence of taeniosis among patients with neurocysticercosis is related to severity of infection. TheCysticercosis Working Group in Peru. PMID- 11061276 TI - Rituximab for myasthenia gravis developing after bone marrow transplant. PMID- 11061277 TI - Cerebral air embolism after central venous catheter removal. PMID- 11061278 TI - Is fluid-attenuated inversion recovery MRI more sensitive than conventional MRI for hypoglycemic brain injury? PMID- 11061279 TI - HMG-CoA reductase inhibitors (statins): a promising approach to stroke prevention. PMID- 11061280 TI - A randomized controlled trial of prednisone in Alzheimer's disease. PMID- 11061281 TI - Use of "unidentified bright objects" on MRI for diagnosis of neurofibromatosis 1 in children. PMID- 11061282 TI - Combined assessment of tau and neuronal thread protein in Alzheimer's disease CSF. PMID- 11061283 TI - Neurologic side effects in neuroleptic-naive patients treated with haloperidol or risperidone. PMID- 11061284 TI - Genomic organization of the human CD5 gene. AB - CD5 is a member of the family of receptors which contain extracellular domains homologous to the type I macrophage scavenger receptor cysteine-rich (SRCR) domain. Here, we compare the exon/intron organization of the human CD5 gene with its mouse homologue, as well as with the human CD6 gene, the closest related member of the SRCR superfamily. The human CD5 gene spans about 24.5 kb and consists of at least 11 exons. These exons are conserved in size, number, and structure in the mouse CD5 homologue. No evidence for the biallelic polymorphism reported in the mouse could be found among a population of 100 individuals of different ethnic origins. The human CD5 gene maps to the Chromosome (Chr) 11q12.2 region, 82 kb downstream from the human CD6 gene, in a head-to-tail orientation, a situation which recalls that reported at mouse Chr 19. The exon/intron organization of the human CD5 and CD6 genes was very similar, differing in the size of intron 1 and the number of exons coding for their cytoplasmic regions. While several isoforms, resulting from alternative splicing of the cytoplasmic exons, have been reported for CD6, we only found evidence of a cytoplasmic tailless CD5 isoform. The conserved structure of the CD5 and CD6 loci, both in mouse and human genomes, supports the notion that the two genes may have evolved from duplication of a primordial gene. The existence of a gene complex for the SRCR superfamily on human Chr 11q (and mouse Chr 19) still remains to be disclosed. PMID- 11061285 TI - Cloning, sequencing, and analysis of expression of a second IL-1beta gene in rainbow trout (Oncorhynchus mykiss). AB - The full-length sequence of a second IL-1beta gene (IL-1beta2) in rainbow trout (Oncorhynchus mykiss) has been obtained. As with the first IL-1beta gene, IL 1beta2 is organized into six exons/five introns. There are only small differences in their intron/exon sizes, with the exception of intron 3, which is 334 bp smaller in IL-1beta2. The transcript encoded by the IL-1beta2 gene contains a 5' untranslated region (UTR) of 121 bp, followed by a 762-bp open reading frame and a 518-bp 3'UTR. The 3'UTR contains seven instability attta motifs, typical of inflammatory genes, and a polyadenylation site 11 bp upstream of a 17-bp poly(A) tail. The predicted 254 amino acid sequence of the second IL-1beta gene has 82% similarity to the first gene, 45% similarity to carp IL-1beta, and 40% similarity to human IL-1beta. Comparison of the two trout genes reveals that the IL-1beta2 gene has a deletion of 9 bases in exon 3 and an altered splicing site at the 5' end of exon 4 giving rise to a further 9-bp deletion in the resulting cDNA. As with other nonmammalian IL-1beta genes, no interleukin-converting enzyme (ICE) cut site has been found but the alignment of the amino acid sequence with other species shows a possible cut site between Arg89 and Ala90 that would give arise to a 165-amino acid mature peptide. Expression studies performed by RT-PCR using primers specific for the IL-1beta2 transcript revealed a clear dose-dependent induction of this gene in cultured trout leukocytes by stimulation with lipopolysaccharide. PMID- 11061286 TI - CD3epsilon homologues in the chondrostean fish Acipenser ruthenus. AB - CD3epsilon is an essential component of the T-cell receptor (TCR) complex for antigen. We report here molecular cloning and characterization of cDNAs encoding the CD3epsilon homologues in sterlet (Acipenser ruthenus), a representative of primitive chondrostean fishes. Sequence analysis of the cDNA clones demonstrated unexpectedly high CD3epsilon gene heterogeneity in this species. While some cDNAs encoded proteins with the structure typical of mammalian CD3epsilon, others coded for proteins lacking the membrane-proximal half of the extracellular domain. Two cDNAs contained in-frame stop codons in the region encoding the cytoplasmic domain. Based on genomic blot analysis and RT-PCR typing of individual spleen RNAs, we suggest that sterlet may possess two highly polymorphic CD3epsilon loci, of which one can produce alternatively spliced transcripts. The structural elements shown to be functionally important in the mammalian CD3epsilon are strongly conserved in the sterlet CD3epsilon. The cytoplasmic region contains an immunoreceptor tyrosine-based activation motif (ITAM) with YEPI and YSGL tyrosine containing sequences that are characteristic of only this TCR subunit. The pattern of sequence conservation indicates also that strong selection pressure was imposed on a motif VYYW at the C-end of the transmembrane domain and on a CD3epsilon-specific proline-rich motif RXPPVP juxtaposed to the N-terminus of the ITAM. Weak similarity of the sterlet CD3epsilon with the chicken and Xenopus CD3gamma/delta indicates that these two TCR subunits diverged before radiation of bony fishes and tetrapods. While the role of CD3epsilon heterogeneity in sterlet remains to be elucidated, the data obtained show that the basic mechanisms of TCR signaling have ancient evolutionary origin. PMID- 11061287 TI - Expression of SpC3, the sea urchin complement component, in response to lipopolysaccharide. AB - The homologue of the vertebrate complement component C3 that is expressed in the coelomocytes of the purple sea urchin, Strongylocentrotus purpuratus, designated SpC3, was investigated for changes in response to immune challenge or injury. Immunoquiescent animals were used in this study because they have reduced or no detectable SpC3 in their coelomocytes or coelomic fluid (CF). Animals were injected with lipopolysaccharide (LPS) or sterile sea water (SSW, injury control). Changes in the amounts of SpC3 in coelomic fluid and in coelomocytes were then followed over time by Western blots and ELISA. Changes in mRNA from the SpC3 gene (Sp064) were also followed by RT-PCR. Although all animals responded to injury with increased levels of SpC3 in the coelomic fluid, those challenged with LPS had greater amounts of SpC3 in both CF and coelomocytes than those receiving SSW. In most of the animals receiving LPS, initial increases in SpC3 were observed within 1 h post-injection, while the earliest response in the animals receiving SSW was 6 h. The appearance of SpC3 in the coelomocytes was delayed compared to its appearance in CF, and was first detected several days after challenge. Changes in mRNA from the Sp064 gene paralleled the appearance of SpC3 in the coelomic fluid. Increases in the number of coelomocytes per milliliter of CF and in the percentage of coelomocytes that were SpC3+ also occurred after challenge with LPS or in response to injury, with a slightly greater increase in response to LPS. Although the changes in SpC3 were not as great as those identified previously for human C3 expressed in macrophages, the kinetics of the response are similar to that of acute-phase reactants in mammals. PMID- 11061288 TI - SpC3, the complement homologue from the purple sea urchin, Strongylocentrotus purpuratus, is expressed in two subpopulations of the phagocytic coelomocytes. AB - The lower deuterostomes, including the echinoderms, possess an innate immune system that includes a subsystem with similarities to the vertebrate complement system. A homologue of the central component of this system, C3, has recently been identified in the purple sea urchin, Strongylocentrotus purpuratus, and is called SpC3. We determined previously that coelomocytes specifically express the SpC3 gene (Sp064); however, the sea urchin has at least four different types of coelomocytes: amoeboid phagocytes, red spherule cells, colorless spherule cells, and vibratile cells. To determine which of these subpopulations expresses Sp064 and produces SpC3, coelomocytes were separated by discontinuous gradient density centrifugation. Relatively homogenous fractions were obtained consisting of the four major cell types in addition to two types of amoeboid phagocytes with different densities and distinct morphologies. Analysis of proteins from separated cell subpopulations by Western blot and analysis of gene expression by RT-PCR revealed that phagocytes express the gene and contain the protein. Immunolocalization showed that SpC3+ phagocytes are present as subsets of both the low- and high-density subpopulations of phagocytes; however, the subcellular localization of SpC3 is different in these two subpopulations. PMID- 11061289 TI - The HLA-DRB4 gene is present in half of the Spanish HLA-DQ2-negative celiac patients. AB - We studied nine consecutive DQ2-negative celiacs [from a group of 186 consecutive celiac disease (CD) patients] for the presence of the HLA-DQB1, DRB1, and DRBx alleles. HLA-DR53 was present in only 5 out of 9 (55%) of DQ2-negative patients. DRB4 (DR53) positivity -39% of chromosomes--among Spanish DQ2-negative CD patients is due to both DR4- and DR7-positive cases. Spanish DQ2-positive patients show a high frequency of DR7/DR11 heterozygous carriers of DRB4 (DR53). One-third of our DQ2-negative celiac patients have DRB1*04 (DR4). Six patients are DR4 negative: at least one of the DQ2 alleles (DQA1*0501/DQB1*02) is present in four cases, but none of the alleles of risk, including DR53, were found in the remaining two cases, both of whom carry DQB1*06 alleles (*0602/3 and 0604). The fact that half of our DQ2-negative patients lack DRB4 (DR53) leads us to believe that this gene is not an essential factor to confer CD susceptibility. PMID- 11061290 TI - A mutation determining the loss of HLA-A2 antigen expression in a cervical carcinoma reveals novel splicing of human MHC class I classical transcripts in both tumoral and normal cells. PMID- 11061291 TI - A new HLA-Cw allele (Cw*0808) found in a Colombian Mestizo individual possibly generated by an intralocus/interloci gene conversion. PMID- 11061292 TI - Arthroscopic management of septic arthritis. PMID- 11061293 TI - Dynamic stability after ACL injury: who can hop? AB - Single-leg hops are used clinically to assess knee function in patients following anterior cruciate ligament (ACL) rupture and reconstruction. Researchers study ACL-deficient individuals in order to identify movement strategies in the absence of a major knee stabilizer, thereby providing information to clinicians regarding treatment options. Single-leg hops represent an activity which places higher demands on the knee than walking or jogging. Hops are thought by some to represent demands that are more comparable to those found during high level sports. Therefore hopping might provide more information about knee stability during dynamic activities than less strenuous activities. This paper reflects one component of a larger study involving comparisons of joint motions and muscle activity patterns in uninjured individuals (n=10) and two groups of athletes who had complete ACL ruptures; one group had substantial knee instability (noncopers, n=10), and the other had no signs of knee instability (copers, n=11). In this paper we report the findings from the single-leg hop activity. The results indicate that coper subjects move in a manner nearly identical to uninjured persons. Kinetic data suggest that copers stabilize their knees with greater contributions from the ankle extensor muscles. Muscle activity data demonstrate that there is no reduction in quadriceps femoris activity in the coper subjects. In the group of ten subjects with knee instability (noncopers) who participated in the overall study involving walking, jogging, hopping, and a step activity only four were willing to hop. Work in our laboratory has established that when high level athletes rupture their ACL, the majority of them cannot return to high level sports. The small number of noncopers in this study who were willing to hop supports our previous findings. Those noncopers who did hop displayed reduced knee range of motion and external knee flexion moments, a movement strategy remarkably similar to that found during other activities. Neither the copers nor the noncopers showed evidence that quadriceps activation was diminished. PMID- 11061294 TI - Arthroscopic management of septic arthritis: stages of infection and results. AB - Seventy-six patients with septic arthritis (78 affected joints) were treated with a combination of arthroscopic irrigation, debridement, and antibiotic therapy according to the tested bacterial sensitivity. There were 62 knee, 10 shoulder, 5 ankle joints, and 1 hip joint. No antibiotics were added to the irrigating solution. The arthroscopic and radiological stage of infection, treatment, and outcome in these patients was analyzed. The patients were classified into three groups according to initial stage of joint infection (stage I: 21 patients, 22 joints; stage II: 43 patients, 44 joints; stage III: 12 patients, 12 joints). Causes of infection were: hematogenous dissemination in 54%, postoperative wound infection in 28% (17% after open, 11% after arthroscopic procedures). Other causes were: 10% intra-articular steroid injections, 3% diagnostic punctures, and 3% open traumatic injury of the joint. In 78% of the infected joints the causative organism could be identified: Staphylococcus aureus was the most common organism found (42%), followed by streptococci (15%), pneumococci (6%), Escherichia coli (4%), Staphylococcus epidermidis (3%), Borrelia burgdorferi (3%), and others in 5%. In the stage I group only one patient needed repeated arthroscopic irrigation, in the stage II group 52%, and in the stage III group 75%. Open revision for eradication of the infection was necessary in one joint with stage II and in two joints with stage III infection (3%). Two joints of the stage III group needed additional surgery after successful treatment of the infection. The combination of arthroscopic irrigation and systemic antibiotic therapy was able to cure 91% of the affected joints. Open revision was necessary in 4% of joints. The number of arthroscopic procedures and the efficacy of treatment depended on the initial stage of the infection. It is concluded that an arthroscopic staging of the initial joint infection has prognostic and therapeutic consequences. PMID- 11061295 TI - Evaluation of harvested and normal patellar tendons: a reliability analyses of magnetic resonance imaging and ultrasonography. AB - This study compared the reliability (interchangeability) of magnetic resonance imaging (MRI) and ultrasonography (US) examinations of the patellar tendon after using central third patellar tendon autografts during anterior cruciate ligament reconstruction. Nineteen consecutive patients (7 women, 12 men) underwent bilateral MRI and US of the patellar tendons 27 (24-29) months after anterior cruciate ligament reconstruction using ipsilateral central third patellar tendon autografts. Two experienced radiologists blinded to one another evaluated the examinations. Measurements of the length of the noninjured patellar tendon showed the greatest reliability between MRI and US, with no systematic difference (P=0.48), a small mean difference (-0.1 mm), and an interclass correlation coefficient of 0.74. The measurements of the thickness and width of the noninjured side were also judged as reliable. However, on the injured side a lower reliability was found between MRI and US. We conclude that MRI and US are reliable (interchangeable) methods only for evaluating noninjured patellar tendons. PMID- 11061296 TI - Type V collagen is increased during rabbit medial collateral ligament healing. AB - To understand the reparative process of medial collateral ligament (MCL), fibrillar collagen and their relative ratios in healing MCL with anterior cruciate ligament (ACL) reconstruction were analyzed. Skeletally mature New Zealand white rabbits were subjected to a mop-end tear of MCL without repair with ACL reconstruction. Rabbits were killed 6 and 52 weeks after injury. Ligamentous tissues from the injury site and sham controls were soaked in 0.5 M acetic acid for 24 h, minced, and treated with pepsin to solubilize collagen. Pepsin solubilized about 80% of the total collagen as determined by hydroxyproline analysis of the pepsin residues. Sodium dodecyl sulfate polyacrylamide gel electrophoresis analysis of the solubilized collagen revealed presence of fibrillar collagen types I, III, and V. Densitometric scanning of the protein bands corresponding to types I, III, and V collagen indicated that in sham controls types III and V collagen represented about 8% and 12%, respectively, of the type I collagen whereas the healed MCL ligaments at 6 weeks showed significant increase in type III and V collagen to about 19% and 24%, respectively. By 52 weeks type III collagen in the healed MCL had returned to that of sham controls while type V collagen remained elevated at approximately 18%. These data suggest that presence of type V collagen in high concentration in healing ligaments may have an influence on collagen fibril diameters seen in healed ligament and should be included in the analysis when evaluating ligament healing. PMID- 11061297 TI - Anterior knee symptoms after four-strand hamstring tendon anterior cruciate ligament reconstruction. AB - Proponents of hamstring anterior cruciate ligament (ACL) reconstruction suggest that anterior knee symptoms (AKS) may be less than following the use of bone patella-bone autograft. Our aim was to assess the incidence of AKS in a cohort of patients who had undergone hamstring reconstructions. Forty-four of 50 consecutive patients who had undergone arthroscopically assisted four-strand gracilis/semitendinosus hamstring ACL reconstructions were reviewed at a minimum follow-up of 24 months. The frequency and severity of anterior knee pain experienced during activities of daily living, sports, prolonged sitting, stair climbing and kneeling was recorded by means of the Shelboume and Trumper anterior knee pain questionnaire. The location of both pain and any perceived sensory change was recorded using patient-drawn diagrams. Although mild or moderate symptoms occurred in a proportion of patients, only 2% experienced significant symptoms that caused limitation with daily activity, 7% with strenuous work or sport, 12% with kneeling, 5% with stairs and none with prolonged sitting. The pain was not specifically related to the incision for tendon harvest and drilling of the tibial tunnel. Areas of sensory change over the front of the knee were identifiable in 50% of patients, and of these, 86% demonstrated sensory change in the distribution of the infragenicular branch of the saphenous nerve. Although rarely a cause of limitation of activity, AKS can be a problem after hamstring ACL reconstruction and patients should be counselled accordingly. PMID- 11061298 TI - Revision ACL reconstruction using autogenous patellar tendon graft. AB - This retrospective study examined revision anterior cruciate ligament reconstruction using a bone-tendon-bone autograft of the patellar ligament. We followed up 44 patients (mean age 27.9 years) for an average of 41.2 months. Clinical examination with the Lachmann and pivot shift tests showed clearly improved stability; KT-1000 arthrometer measurements had a mean difference of 3.5 mm in side-to-side comparison. The evaluated knee scores were significantly improved (P<0.01); the median Lysholm score was 85 and the median Tegner activity score 5.0 at follow-up. In the IKDC ranking system 75.0% of knees were rated normal or nearly normal (grades A and B). According to a modified Fairbank scale, progression of radiographic signs of osteoarthritis was noted in 36.4%. There was a significant difference (P<0.05) in progression of radiographic signs of osteoarthritis between patients with major (grades III, IV) versus minor (grades I, II) lesions of the articular cartilage surface and between knees with versus without extensive synovitis due to previous synthetic graft reconstruction (P<0.05). Revision anterior cruciate ligament reconstruction using an autogenous patellar tendon graft shows good results with improved knee function compared to the prerevision status and is in line with various operative techniques described in the literature. Progression of osteoarthritis must be expected in patients with major lesions of the articular cartilage surface and knees with long-term extensive synovitis due to previous anterior cruciate ligament reconstruction using synthetic grafts. PMID- 11061299 TI - Motor control performance in the lower extremity: normals vs. anterior cruciate ligament reconstructed knees 5-8 years from the index surgery. AB - We compared motor control function in 50 patients who had undergone anterior cruciate ligament reconstruction using a bone-tendon-bone graft to that in 50 normal controls. Surgical subjects patients had undergone reconstruction with a one- (n=37) or two-incision (n=13) technique with the same rehabilitation protocol; mean time from the index surgery was 6.1 years (range of 5-8 years). For inclusion patients required an excellent outcome, category A IKDC score, and a KT-1000 side-to-side difference of 3 mm or less. Motor control evaluations were conducted using the KAT 2000 with static and dynamic tests. Normal controls had substantially better scores than did the surgical patients. There was no statistical difference the single-limb static test between scores of operated and nonoperated limbs. However, the operated limb scores were slightly better overall than those for the nonoperated limb, and the right knee scores tended to be better than those for the left knee. This may be explained by limb dominance. The test method employed in this investigation shows that anterior cruciate ligament reconstructed patients had a clear motor control deficit compared to normal control subjects even after several years. PMID- 11061300 TI - Isokinetic torque deficit of the knee extensor muscles after arthroscopic partial meniscectomy. AB - Isokinetic torque deficit of the knee extensor muscles in the operated leg was measured in 21 male patients (mean age 26.4 +/- 1.9 years) who had undergone arthroscopic partial medial meniscectomy. The isokinetic torque testing was performed 1, 3, and 6 months postoperatively using the Cybex II dynamometer according to standard technique. Isokinetic knee extension peak torque (PT) at angular velocities of 60 degrees and 180 degrees/s was determined in both legs, and the proportional PT deficit in the operated leg was compared with that in the nonoperated leg. A significant (P<0.001) isokinetic PT deficit in the operated leg in testing with angular velocity of 60 degrees and 80 degrees/s was observed at 1 month (28.6% and 31.0%, respectively) and 3 months (19.8% and 15.8%, respectively) postoperatively. At 6 months postoperatively a significant (P<0.001) isokinetic PT deficit (18.2%) of the knee extensor muscles in the operated leg was observed only in testing with angular velocity of 60 degrees/s; no significant differences (P>0.05) in isokinetic PT between the operated and nonoperated leg in testing with angular velocity of 180 degrees/s was found 6 months postoperatively. Thus in patients with arthroscopic partial meniscectomy the postoperative recovery of isokinetic strength of the knee extensor muscles in the injured leg is closely related to testing velocity, while it is more delayed at low than intermediate angular velocities. PMID- 11061301 TI - Prolonged peroneal nerve dysfunction after high tibial osteotomy: pre- and postoperative electrophysiological study. AB - To evaluate electrophysiological incidence and the type of peroneal nerve lesions seen after high tibial osteotomy we conducted an electrophysiological study (electromyography and nerve conduction velocity studies) in 11 patients who were suffering from medial gonarthrosis and treated by Maquet barrel-vault type high tibial valgization osteotomy. All the patients were tested both pre- and postoperatively. Every patient was examined postoperatively for a minimum of a 6 months after surgery to eliminate spontaneously reversible lesions. Results obtained from nonoperated legs served as controls. Three patients (27%) with peroneal nerve lesions were detected electrophysiologically; one had only motor involvement, one only sensory involvement, and one both motor and sensory involvement. Clinically only one of these patients was symptomatic, and the other two were detected by electrophysiological means. Peroneal nerve lesions which may be overlooked by mild weakness and hypesthesia in the early postoperative period can be detected by electrophysiological means at a higher rate than expected. These lesions persist a relatively long time and even can be permanent despite the absence of clinical symptoms. PMID- 11061302 TI - The coracoacromial ligament: anatomical and biomechanical properties with respect to age and rotator cuff disease. AB - The coracoacromial ligament (CAL) plays an important role in the pathoetiology of the subacromial impingement syndrome especially in those patients who do not have bony abnormalities. A total of 40 shoulders were dissected to determine the anatomical and biomechanical properties of the CAL in shoulders with either intact rotator cuffs or rotator cuff disease, taken from cadavers of persons who were of various ages at death. The specimens from cadavers with rotator cuff degeneration had a shorter lateral and medial band of the CAL than those of the specimens taken from shoulders with intact rotator cuffs. The cross-sectional area of the lateral band was also enlarged in older specimens with rotator cuff degeneration. Analysis of the structural properties showed a higher load to failure and a higher stiffness in the younger than in older specimens. In material properties, there was a higher failure stress in specimens with normal rotator cuffs than in the specimens with rotator cuff disease but only in older specimens. The decreased material properties in older specimens with rotator cuff disease may be caused by the previously reported histological differences with tissue disorganization and a lack of parallel bundle orientation associated with rotator cuff disease. PMID- 11061303 TI - Results of the original Putti-Platt procedure for shoulder instability: review of Putti's scholar experience. AB - Twenty patients who underwent Putti-Platt shoulder capsulorraphy were retrospectively evaluated at long-term follow-up (24-34 years). The aim of this study was to analyse the efficacy and possible degenerative changes associated with this procedure. Patients' charts were reviewed to analyse the pre-operative picture, surgical technique and post-operative program. All patients were re examined using ASES, Rowe and Lysholm scales. Range of movement (ROM) of the shoulder operated on was compared with the non-operated one. A-P, axillary and outlet views were taken for radiographic control. Samilson criteria were followed to determine the degree of arthrosis. Clinical outcome was satisfactory in 85% of the cases with only three cases with fair results (15%). No recurrence was present in this series. The mean limitation of external rotation in abduction was 9 degrees with respect to the contra-lateral shoulder. Severe degenerative changes were observed in two cases. In conclusion, after a long follow-up period, the Putti-Platt technique has shown highly satisfactory results, with an incidence of loss of motion and joint degeneration changes similar to other procedures. PMID- 11061304 TI - Peroxidative degradation of selected PCB: a mechanistic study. AB - The enzyme-induced decomposition and biodegradation of PCB were investigated. 2,5 Dichlorobiphenyl (PCB 9) and 2,2',5,5'-tetrachlorobiphenyl (PCB 52) were used as example compounds to study efficiency and mechanism of the degradation processes. It was found that the application of horseradish peroxidase (HRP) together with defined amounts of hydrogen peroxide removed 90%, of the initial concentration of PCB 9 and 55% of the initial concentration of PCB 52 from an aqueous solution after a reaction period of 220 min. Dechlorination was observed as the initial step. Although the metabolites identified were mainly chlorinated hydroxybiphenyls, benzoic acids and non-substituted 1,1'-biphenyl, some higher chlorinated biphenyl isomers also appeared. The biodegradation of PCB 9 using the white rot fungus Trametes multicolor took about four weeks and reduction was about 80%, of the initial concentration. The metabolites produced (dichlorobenzenes, chlorophenols and alkylated benzenes) were not quite the same as those observed upon incubation with HRP. PMID- 11061305 TI - Remediation of s-triazines contaminated water in a laboratory scale apparatus using zero-valent iron powder. AB - Atrazine, propazine and simazine were tested separately and in mixture by batch procedure in a laboratory-constructed apparatus. 3.75 l of a buffered s-triazines pesticide solution was treated at room temperature by 325-mesh zero-valent iron powder (ZVIP) (20 g/l). High performance liquid chromatography was used to separate by-products and study the decline in the pesticide's concentrations. Results obtained show that the order of degradation was simazine, atrazine and then propazine. The half-lives (t1/2) of the s-triazines pesticides are, respectively, 7.4, 9.0 and 10.6 min when they are treated separately, and 9.8, 11.2 and 13.7 min when they are treated together under the same conditions. The final by-product obtained after 50 min of contact of simazine with ZVIP shows a shift to longer wavelength in its UV spectrum. A similar phenomenon is shown for atrazine and propazine. Identical primary by-products are produced and subsequently degraded to 4,6-(diamino)-s-triazine, which seems to be the major by product of the reductive treatment process. Pathways for the degradation of the studied s-triazines by ZVIP are proposed. PMID- 11061306 TI - Polycyclic aromatic hydrocarbons in the bulk precipitation and surface waters of Northern Greece. AB - Bulk (wet and dry) precipitation and surface water sampling was undertaken in the main plain of central Macedonia in Northern Greece. Fourteen polycyclic aromatic hydrocarbons (PAHs) included in the US EPA's priority pollutant list were analysed. The concentrations determined in bulk precipitation were in general within the range of values worldwide reported. Concentrations were highest in the cold months. Deposition fluxes of PAHs were of the same order of magnitude as reported data. The greatest values were found when high concentrations of PAHs in precipitation coincided with large precipitation amounts. The concentrations of PAHs in surface waters (main rivers, tributaries, ditches, etc) were in general lower than those in bulk precipitation, and among the lowest reported for European rivers, excepting Np and Ph. Bulk deposition and domestic effluents are suggested as being the main PAH sources into surface waters. PMID- 11061307 TI - Characteristics of sorption losses of polychlorinated biphenyl congeners onto glass surfaces. AB - Sorption losses to glass surfaces of five polychlorinated biphenyl (PCB) congeners in aqueous solutions were investigated. Adsorption/desorption experiments were conducted under conditions that simulated actual sample handling procedures for environmental samples. It was found that the adsorption loss is related to the degree of chlorination. PCB congener 180 lost the most onto glass surfaces, followed by congeners 138, 101/28, and 52, in decreasing order. More PCB adsorption occurred onto glass under conditions of agitation and higher temperature (22 degrees C) during the five-day experimental period. The salinity effect ("salting out effect") was also observed in this work. The efficiency of desorption (rinsing three times with solvent) was found to be ineffective in extracting adsorbed PCBs. It was necessary to use mechanical shaking for extraction. Storage of samples up to five days resulted in sorption losses as much as 30%, 17%, 30%, 40%, and 55% of PCB 28, 52, 101, 138, and 180, respectively. Sorption losses need to be considered when conducting water sampling or toxicological studies to avoid underestimation of the actual PCB concentrations and their toxic effects. PMID- 11061308 TI - Coating effects on the glass adsorption of polychlorinated biphenyl (PCB) congeners. AB - The effects of coating materials on polychlorinated biphenyl (PCB) adsorption in aqueous solution were assessed in an attempt to minimize PCB sorption loss during sampling processes. A coating material, which enhances PCB adsorption and allows adsorbed PCBs to be readily extracted by solvents, can act as a sampling concentrator to reduce PCB losses from both adsorption and evaporation. Several coating materials were evaluated, including paraffin oil, silicone oil, dimethyldichlorosilane (Sylon-CT), Prosil 28 and polydimethylsiloxane (PDS) with viscosity 0.65, 50 (PDS 50), and 500 (PDS 500) cSt. PDS and silicone oil enhanced adsorption for all five congeners examined (IUPAC No. 28, 52, 101, 138, and 180). Sylon-CT, paraffin oil and Prosil 28 had inconsistent effects on adsorption of different congeners. Desorption of adsorbed PCBs onto all coating types was assessed. The recovery efficiency of extracting PCBs with solvents was enhanced greatly with all coatings as opposed to non-coated surfaces, with the exception of paraffin oil. Coating with silicon oil, PDS 50, and 500 resulted in virtually 100% recovery of adsorbed PCBs. It was also found that Teflon containers were poor substitutes for glass containers and failed to minimize PCB losses. Among the materials studied, the best coating that could be used as a sampling concentrator was PDS 500. PMID- 11061309 TI - Biodegradation of 2,4,6-trichlorophenol in the presence of primary substrate by immobilized pure culture bacteria. AB - In this study, pure strains that are capable of utilizing 2,4,6-trichlorophenol have been isolated from the mixed culture grown on substrates containing chlorophenolic compounds. Studies have been carried out on the capability of these isolated pure strains in suspended and immobilized forms to decompose 2,4,6 trichlorophenol. Additionally, the influence of primary substrates (e.g., phenol, 2-chlorophenol, 3-chlorophenol, 4-chlorophenol, 2,4-dichlorophenol) on the decomposition of 2,4,6-trichlorophenol by the isolated pure strains grown in immobilized form is also investigated. The results are: Through bacterial isolation and identification, three pure strains have been obtained: Pseudomonas spp. strain 01, Pseudomonas spp. strain 02 and Agrobacterium spp. Whether in suspended or immobilized forms, all strains have poor removal efficiencies of 2,4,6-trichlorophenol. However, addition of 200 mg/l phenol will enable the immobilized Pseudomonas spp. strain 01, and Pseudomonas spp. strain 02 to achieve 65% and 48% removal of 2,4,6-trichlorophenol, respectively. Addition of phenol will assist the immobilized Pseudomonas spp. strain 02 in achieving removal of 2,4,6-trichlorophenol but the removal efficiency is not good if the phenol concentration is too low. The optimum phenol concentration should be between 200 and 400 mg/l. PMID- 11061310 TI - Dechlorination of polychlorinated biphenyls, dibenzo-p-dioxins and dibenzofurans on fly ash. AB - Dechlorination of commercial mixtures of polychlorinated biphenyls (PCB) as well as polychlorinated dibenzo-p-dioxins (PCDD) and dibenzofurans (PCDF) on extracted and non-extracted fly ash obtained from municipal waste incinerator (MWI) was studied in closed systems under nitrogen atmosphere at temperatures of 260 degrees C and 340 degrees C. Decomposition results (given as the difference between PCB or PCDD/F molar amounts before and after the experiment (in %) due predominantly to dechlorination reactions) and detoxification data (expressed similarly but related to toxic PCB and PCDD/F congeners only and given in I-TEQ units) are reported. Detoxification of Delor 105/80T at 260 degrees C and 340 degrees C at a loading of 0.65 wt%, was 99.48% and 100%, respectively. The decomposition of Delor 103 at 340 degrees C and for the loading of 0.75 wt%, corresponded to 99.99%. The detoxification capability of PCDD/Fs on extracted and non-extracted fly ash for loading of 130 and 264 ng/0.4 g of fly ash at 340 degrees C made 96 and 98%, respectively. PMID- 11061311 TI - A new method of predicting of gas chromatographic retention indices for polychlorinated dibenzofurans (PCDFs). AB - A new method has been developed to describe the quantitative relationship between molecular structures of PCDFs and their gas chromatographic retention indices on a 30-m fused silica column coated with DB-5 stationary phase. The regression equation is derived with a multiple correlation coefficient greater than 0.9995. The highest residual is 20 index units. The standard deviation is less than 7 index units. Using this regression equation, the retention indices of PCDFs for which data is not available have also been predicted. PMID- 11061312 TI - Polychlorinated organic compounds in Yangtse River sediments. AB - Polychlorinated organic compounds (PCOCs) were analyzed in Yangtse River sediments. The results show that the concentrations of PCOCs in Yangtse River sediments followed the order of DDTs > HCB > HCHs > PCBs. High PCOCs concentrations were detected in sediments from station Y02a and Y02b, which are located in the main input of the Yangtse River (Nanjing section). Results also show that the PCOCs values were highly correlated with organic carbon content and heavy metal contamination. PMID- 11061313 TI - The effect of oils on PAH, PCDD, PCDF, and PCB emissions from a spark engine fueled with leaded gasoline. AB - The effect of synthetic and mineral oils on the formation of polyaromatic hydrocarbons (PAHs), polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs), and biphenyls (PCBs) in emissions from a spark ignition engine was studied on a Skoda Favorit engine fueled with leaded gasoline. The test cycle simulated urban traffic conditions on a chassis dynamometer, in accordance with the ECC 83.00 test. The data for selected PAHs as well as PCDDs, PCDFs, and PCBs congener profiles are presented. PCDD/Fs emissions for an unused oil and the oil after 10000-km operation varied from 300 to 2000 fmol/m3, PCBs emissions from 75 to 178 pmol/m3, and PAHs emissions from 150 to 420 microg/m3. The content of PCBs in oils varied from 2 to 920 mg/kg. PMID- 11061314 TI - Tissue distribution of polychlorinated naphthalenes (PCNs) and non-ortho chlorinated biphenyls (non-ortho CBs) in harbour porpoises (Phocoena phocoena) from Swedish waters. AB - Polychlorinated naphthalenes (PCNs) and non-ortho chlorinated biphenyls (non ortho CBs) were analysed in blubber, nuchal fat, liver, muscle, kidney and brain of three male harbour porpoises (Phocoena phocoena) from the west coast of Sweden. To estimate spatial variation, PCNs and non-ortho CBs were analysed in six blubber samples collected at different anatomical sites of each animal. Highest wet weight concentrations of sigma PCNs were detected in the lipid rich tissues (blubber and nuchal fat) and liver (520-730 and 520 pg/g, respectively) and lowest in brain (22 pg/g). TetraCNs were most abundant in muscle, kidney and brain, while the hexaCNs were most abundant in the lipid rich tissues and liver. The highest lipid weight concentration recorded (11 ng/g) was for the hexaCN congeners no. 66/67 in liver. These coeluting hexaCN congeners accounted for 80 100% of total hexaCNs in all tissues examined. Concentrations of sigma non-ortho CBs were highest in lipid rich tissues (220-280 pg/g wet weight). Non-ortho CB no. 77 and 169 constituted between 62-86% and 4.9-9.3%, respectively, of total sigma non-ortho CBs. No major variation of sigma non-ortho CB concentrations was found between the six different blubber sites but higher sigma PCN concentrations (wet weight) were found dorsally at the peduncle. Toxic equivalent concentrations (TEQs) showed that non-ortho CB no. 126 was the main contributor to total TEQs in all tissues, except liver in which hexaCN congener nos. 66/67 contributed to about 50% of total TEQs. PMID- 11061315 TI - Analysis of dioxin-like compounds in vegetation and soil samples burned in Catalan forest fires. Comparison with the corresponding unburned material. AB - Only a few data are reported about the formation of polychlorinated dibenzo-p dioxins (PCDD) and dibenzofurans (PCDF) in forest fires. However, the inventories of sources undertaken by several European and American countries consider natural fires as a possible source of PCDD/F and, in some cases, as one of the most important. In our work we have analysed vegetation and soil samples burned in four forest fires which happened in Catalonia in the summer of 1998. The concentration of dioxin-like compounds (PCDD/F, non-ortho polychlorinated bi phenyls (PCB) and mono-ortho PCB) has been compared to the concentration present in the corresponding unburned material collected in places near the fires but not affected. The results of this preliminary study show very low concentrations in all the samples, both burned and unburned. Although a change in the profile (proportional increase of tetrachlorinated congeners in PCDD/F) is observed in burned samples compared to unburned ones, the absolute values of concentration decrease in most samples. Therefore, natural fires seem not to be an important source of dioxin-like compounds. These results will be confirmed with air emissions measurements in future studies. PMID- 11061316 TI - The scholar I. Athanasiu and his scientific work. AB - The personality of the scientist I. Athanasiu united the happy complementarity of an exceptional professor and researcher. If Professor I. Athanasiu perceived science by its past and transmitted it in a synthetized form to his contemporaries, the researcher I. Athanasiu was permanently interested in the future, in the following stages of scientific research. In his short life, Professor I. Athanasiu illustrated brilliantly both aspects of a scientist's personality. Adding these qualities to his vivid, nonalterated and discrete patriotism, shown honestly among foreigners in the major European countries as well as among Romanians and foreigners in his country, his worries for the future of his homeland, especially for its youth, we realise another aspect of I. Athanasiu's personality, that of a scientist and patriot. By his scientific research, I. Athanasiu surpassed the limits of medical-veterinary area, approaching with talent and efficiency its related fields: human medicine and animal biology. PMID- 11061317 TI - Professor Ioan Athanasiu's contributions in the promotion of the graphic and cinematographic method in physiology. PMID- 11061318 TI - Ioan Athanasiu--founder of the new Institute of Physiology at the University of Bucharest. PMID- 11061319 TI - Professor Ioan Athanasiu--the main promotor of the Superior School of Veterinary Medicine at the standards of a faculty. PMID- 11061320 TI - Professor Ioan Athanasiu, teacher and protector of university youth. PMID- 11061321 TI - [Professor Ioan Athanasiu and military medicine]. PMID- 11061322 TI - References on Professor I. Athanasiu's contribution to the developement of physiology (a chronology of the works). PMID- 11061323 TI - Newer issues on the topics of stress and of exercise physiology. AB - The background concerns the present-day psychobiological construct on stress and its models. Psychoneuroendocrine mechanisms, cellular and molecular alterations, oxidative and immune reactions and stress system disorder, related to physical, psychosocial and emotional stress became hot research areas. Other important directions regard occupational stress, objective and subjective assessment methods and stress prevention and management. For the physiology of exercise the investigation of central and peripheral limiting factors of human physical performances and the involvement of neuroscience to exercise studies are of main interest. A commentary of the stress concept in terms of testability (Popper) and of a paradigm (Kuhn) follows. The contributions of the Cluj laboratory to the neuroendocrine control in stress, exercise and work, leading to some practical outcomes, and the connections between psychoanalysis and the stress construct are discussed. PMID- 11061324 TI - The adaption to environmental factors: hazard or programme. AB - Any living structure, from cell to the entire organism, can adapt to internal or external factors only by triggering pre-existing mechanisms. All these mechanisms have a morphological support programmed within foetal inactive genes or adult genes with low activity. These genes are activated by a chain of reactions that usually starts with perceiving the stimulus at the membrane level and finishes with the synthesis of some proteins (enzymes, membrane receptors, etc.). The adaptative changes of the enzymatic and structural register would be impossible without the activation of genes other than those operating in normal condition. It is obvious that the same pre-existing genetic mechanism also functions in myocardial hypertrophy or neoangiogenesis. Actually, "a genetic orchestra" is ready to perform at a given moment the new tone suggested by the ever changing condition under which the myocardial fiber acts. However, the concept of programmed adaption has not only a theoretical value; in medical practice, the prediction, prevention and treatment of ischemic heart disease will benefit from the theoretic support of understanding the cardiac adaptative behaviour. It is obvious that this adaptation programmes cannot always save the heart from necrosis and the subject from death due to fatal arrhythmias or cardiogenic shock. PMID- 11061325 TI - The role of the Gregg phenomenon in cardiac performance. AB - The Gregg phenomenon has not, yet, a full and complete explanation. We are allowed to consider it a "garden house"-like effect. Positive inotropic factors are released as a consequence of "shear stress" phenomenon following an increased coronarian flow. In our study, it is more probably a "garden house"-like effect that causes a sarcomere stretch. This could explain the increased left ventricle performance found, in some circumstances, in our study. We used the data obtained by coronarography in 22 patients carrying various degrees of coronarian obstruction and we assessed the left ventricle performance during the repeated injection of contrast medium, that increases the pressure in the coronarian bed. The results obtained support the theory regarding the Gregg effect but do not allow us to consider this effect as a fundamental determinant of the cardiac performance. PMID- 11061326 TI - The involvement of endogenous opiates in emotional-cognitive interaction in stress. AB - Stress system is very important in the co-ordination of the generalised stress response, which takes place when a stressor of any kind exceeds a threshold. In our experiments we have analysed aspects of opiate involvement in the interactions among the emotional, motivational and cognitive processes, as well as aspects concerning the relations between physiological and behavioural mechanisms, both in normal conditions and under stress. We also analysed aspects of the interaction between stress system and endogenous opioid systems (e.o.s.). As emotional states interfere with perception and cognition, in our experiments we used suitable variants of the open-field test, which evaluates the exploratory drive and emotional reactivity. We used 30 Wistar male rats, 18 months old, weighing 350-400 g, divided into three groups, of 10 animals each. In order to assess the behaviour of the animals, we have analysed exploratory (cognitive) and non-exploratory (emotional) parameters. In order to evaluate the manner in which endogenous opioid systems are involved in these aspects, different groups of rats were treated with an opiate agonist (codeine syrup, 0.117, 5 ml/kg to approximately 0.5 mg/kg morphine as active compound, p.o.) and an antagonist (naloxone 0.4 mg/kg i.p.), before testing in open-field. To distinguish between the effect of habituation and the effect of the opiate agonist and antagonist, the placebo treatment and the test/retest method were used. At these doses, the opiate agonist has stimulated the exploratory behaviour, while the antagonist has inhibited the cognitive behaviour and enhanced the emotional response. Our experimental data together with data from literature allowed us to present an interactive model that points out a general manner of interaction between emotions and cognition, in relation to the interactions between physiological and behavioural mechanisms. PMID- 11061327 TI - Intrinsic neuronal time delays can be compensated in cat visual cortex and frog tectum with regard to motion analysis. AB - Motion detection is an essential biological property of vertebral brain. In order to localize moving objects exactly, intrinsic time delays of the neuronal network must be compensated for. Invariance of position with regard to velocity of a stimulus due to a negative spatial shift is one option for compensation. Experimental results found in the present study support the view that negative spatial shift occurs in the visual cortex of the cat and the tectum of the frog. An order of 30% of the visual neurons may be suited to compensate intrinsic time delays. PMID- 11061328 TI - Research on red cell membrane permeability in arterial hypertension. AB - Arterial hypertension, including the elucidation of hypertension pathogenic mechanisms involving elements in the composition of the blood, continues to represent a topical research area. Recent work, such as nuclear magnetic resonance studies looking into red cell permeability, illustrates the presence of modifications of red cell permeability to water (RCPW) related to the stage of arterial hypertension. The identification of a significant increase of RCPW compared to that present in the population with normal arterial pressure values can be useful both in early diagnosis and in warning about a possible predisposition for this condition. At the same time, the dynamic investigation of protonic relaxation time of both intra- and extra-erythrocytic water, the assessment of proton exchange time across the red cell and the calculation of permeability to water enable one not only to diagnose arterial hypertension but also to ascertain the evolution of the disease, its complications and the effectiveness of anti-hypertensive medication. Our studies have also proven the existence of a correlation between the values of systolic arterial pressure and red cell permeability to water. The curve describing the interdependence of the two values has the shape of a bell, in the case of males. The peak of the curve is reached for a systolic pressure of 160 mmHg and gets below the values of the control group in the case of systolic pressures above 200 mmHg. The RCPW test can also be considered a valuable indicator in evaluating the risk of stroke in hypertensive patients. In the chronic therapy of arterial hypertension with various types of anti-hypertensive drugs, one can note differences in the RCPW values related to the effectiveness of the respective medication, to the clinical form and stage of the disease, the sex of the patient as well as to the existence of cerebro-vascular complications. PMID- 11061329 TI - Activity of Na+/K+-ATPase and of Ca++-ATPase under the action of adenosine triphosphate in experimental myocardial hypertrophy. AB - This paper presents the results of our research on the mechanisms involved in the modifications occurring in the activity of ionic pumps (Na+/K+-ATPase and Ca++ ATPase) at the level of the sarcolemma, the sarcoplasmic reticulum and mitochondrial membrane of the myocardial cell in experimental myocardial hypertrophy induced by isoproterenol. We also studied the effects of concomitant administration of adenosine triphosphate. Thus, we found the activity of the sarcolemmal Ca++-ATPase intensely increased under the action of isoproterenol, while Ca++-ATPase activity in the sarcoplasmic reticulum decreased significantly. At the same time, the Na+/K+-ATPase activity decreased both in the sarcolemma and in the sarcoplasmic reticulum. Administration of adenosine triphosphate induces opposite effects, of lower intensity, upon the activity of the two ATPase, that tend to offset the the effects of isoproterenol. This was found in the group of rats given concomitantly both isoproterenol and adenosine triphosphate. A better understanding of the processes involved in those modifications of membrane ATPase activity allows us to consider their different behaviour to isoproterenol and adenosine triphosphate as the expression of intrinsic mechanisms by means of which the myocardial cell intervenes in maintaining a physiological concentration of calcium ions (Ca++) that is necessary both in order to avoid a failure of the contractile apparatus and in order to preserve the mitochondria role of ATP supplier. Our results demonstrate the antiadrenergic action of adenosine triphosphate that attenuates isoproterenol effects on cardiac myocytes. PMID- 11061330 TI - Influence of bilateral olfactory bulbectomy on the circadian rhythm of phagocytic activity and phagocytic response in mice. AB - The bilateral olfactory bulbectomy (OB) has been proposed as an animal model of depression. The present study, carried out on NMRI adult male mice kept on a natural LD 12:12 regimen, aimed to assess the influence of OB on the circadian rhythm of blood neutrophils phagocytosis. The results show that OB mice present an about 20% reduction of the basal phagocytic activity at 09.00 hrs, 15.00 hrs. and 21.00 hrs. and a 40% reduction at 03.00 hrs., leading to a flattened circadian phagocytic curve. The results indicate that bilateral olfactory bulbectomy depresses phagocytosis, alters its circadian rhythm and consequently increases susceptibility to infections. PMID- 11061331 TI - Circadian rhythm of phagocytosis in mice. AB - There has been reported, in some diurnal or equivocal species (man, respectively guinea pig), a circadian rhythm of the phagocytic activity of blood neutrophils, with an acrophase occurring at the end of the light span. The present study, carried out on NMRI adult male mice kept on a LD 12:12 regimen, aimed to assess any circadian variation in the blood neutrophils' phagocytosis level. Basal phagocytic activity was tested against E. coli, every three hours of a 24 h cycle. The results show that phagocytosis in mice's blood neutrophils also presents a rhythmic circadian variation, whose acrophase is delayed with about eight hours compared to that in man, occurring in the second half of the dark period (3:00 h). The occurrence of high circadian phagocytic levels appears to be correlated with the activity type of the species and, of this point of view, mice cannot be used as a model for chronotherapeutical approaches in humans, without keeping in mind the differences between the time structure of the two species. PMID- 11061332 TI - High glucose concentrations increase the tumor necrosis factor-alpha production capacity by human peripheral blood mononuclear cells. AB - The proinflammatory cytokines tumor necrosis factor-a (TNF) and interleukin-1b (IL-1) play an important role in the pathogenesis of insulin-dependent diabetes mellitus, while TNF is also involved in promoting insulin resistance. It has been recently shown that glucose can induce the synthesis of TNF and IL-6 in human monocytes. The aim of the present study was to investigate the effect of glucose on unstimulated and lipopolysaccharide (LPS)-induced TNF and IL-1 production by human peripheral blood mononuclear cells (PBMC). No effect of glucose on spontaneous release of TNF or IL-I could be observed. The LPS-stimulated production of TNF was enhanced when cells were preincubated with increasing glucose concentrations. In contrast, no effect of glucose preincubation on LPS induced IL-I synthesis was found. In conclusion, high glucose concentrations can increase the stimulated TNF production capacity, with possible important consequences for patients with diabetes mellitus. PMID- 11061333 TI - The effects of dopaminergic mediation upon colloidopectic activity of the mononuclear phagocyte system. AB - The investigations were carried out on 30 Wistar albino male rats. A group of animals received Levodpa (Nakom), another group received Methyldopa (Dopegyt). One and a half hour after administration, the phagocytic activity was measured using colloidal carbon. Subsequent to Levodopa administration a significant increase occurred in the phagocytic activity. Following the treatment with Methyldopa the phagocytic activity in macrophages decreased. Our studies confirm the idea that the central adrenergic system stimulates the colloidopectic capacity of macrophages. PMID- 11061334 TI - Sources on the anterior and posterior banks of the central sulcus identified from magnetic somatosensory evoked responses using multistart spatio-temporal localization. AB - A Multi-Start Spatio-Temporal (MSST) multidipole localization algorithm was used to study sources on the anterior and posterior banks of the central sulcus localized from early somatosensory magnetoencephalography (MEG) responses. Electrical stimulation was applied to the right and left median nerves of 8 normal subjects. Two sources, one on the anterior and one on the posterior bank of the central sulcus, were localized from 16 data sets (8 subjects, 2 hemispheres). Compared with the more traditional practice of single-dipole fits to peak latencies, MSST provided more reliable source locations. The temporal dynamics of the anterior and posterior central sulcus sources, obtained using MSST, showed considerable temporal overlap. In some cases, the two sources appeared synchronous. On the other hand, in the traditional single-dipole peak latency fit approach, there is no time course other than a focal dipole moment activated only at the selected peak latency. The same group of subjects also performed a motor task involving index-finger lifting; the anterior central sulcus source obtained from electrical median nerve stimulation localized to the same or similar region in the primary motor area identified from the finger-lift task. The physiological significance of the anterior central sulcus source is discussed. The findings suggest that one can test the integrity of cortical tissue in the region of primary motor cortex using electrical somatosensory stimulation. PMID- 11061335 TI - Coupling of regional activations in a human brain during an object and face affect recognition task. AB - Magnetic field tomography (MFT) was used to extract estimates for distributed source activity from average and single trial MEG signals recorded while subjects identified objects (including faces) and facial expressions of emotion. Regions of interest (ROIs) were automatically identified from the MFT solutions of the average signal for each subject. For one subject the entire set of MFT estimates obtained from unaveraged data was also used to compute simultaneous time series for the single trial activity in different ROIs. Three pairs of homologous areas in each hemisphere were selected for further analysis: posterior calcarine sulcus (PCS), fusiform gyrus (FM), and the amygdaloid complex (AM). Mutual information (MI) between each pair of the areas was computed from all single trial time series and contrasted for different tasks (object or emotion recognition) and categories within each task. The MI analysis shows that through feed-forward and feedback linkages, the "computation" load associated with the task of identifying objects and emotions is spread across both space (different ROIs and hemispheres) and time (different latencies and delays in couplings between areas)-well within 200 ms, different objects separate first in the right hemisphere PCS and FG coupling while different emotions separate in the right hemisphere FG and AM coupling, particularly at latencies after 200 ms. PMID- 11061336 TI - Quantitative maps of GAbAergic and glutamatergic neuronal systems in the human brain. AB - GABAergic and glutamatergic neuronal systems in adult normal human brains were shown quantitatively and in detail through the distributions of glutamate decarboxylase (GAD) and glutamate dehydrogenase (GDH), respectively. Consecutive coronal sections containing part of the striatum and the substantia nigra were obtained from the right hemisphere of three deceased persons with no history of neurological or psychiatric diseases and were stained immunohistochemically for GAD and GDH. Each stained section was divided into approximately 3 million microareas and the immunohistochemical fluorescence intensity in each area was measured by a human brain mapping analyzer, which is a microphotometry system for analysis of the distribution of neurochemicals in a large tissue slice. In the analyzed brain regions, conspicuously intense GAD-like immunoreactivity was observed in the substantia nigra, globus pallidus, and hypothalamus. GDH was widely and rather evenly distributed in the gray matter compared to GAD, although intense GDH-like immunoreactivity was observed in the lateral geniculate nucleus and substantia nigra. Within the substantia nigra, the globus pallidus, and other regions, characteristic distributions of GAD- and GDH-like immunoreactivity were found. We believe that the analysis of the human brain by this novel technique can help to understand the functional distribution of neuronal systems in the normal human brain and may be able to identify abnormal changes in the diseased human brain. It can also provide basic data to help in the interpretation of functional magnetic resonance imaging or positron emission tomography. PMID- 11061337 TI - Visual exploration of form and position with identical stimuli: functional anatomy with PET. AB - Visual form and position perception in primates is thought to engage two different sets of cortical visual areas. However, the original concept of two functionally different and anatomically segregated pathways has been challenged by recent investigations. Using identical stimuli in the centre of the visual field with no external cues, we examined whether discrimination of form aspects and position aspects would indeed activate occipito-temporal and occipito parietal areas, respectively. We measured and localised regional cerebral blood flow (rCBF) changes in the brain with positron emission tomography (PET) and 15O butanol while the subjects performed four visual tasks: position discrimination (PD), form discrimination (FD), joint form and position discrimination (FPD), and a control task. Discrimination of form contrasted with discrimination of position resulted in rCBF increases in the lateral occipital and fusiform gyri. Discrimination of position contrasted with discrimination of form yielded rCBF increases in the left frontal eye field and middle frontal gyrus. No extra activations were seen when the joint form and position discrimination task was contrasted with either the individual form and position discrimination tasks. When the individual form and position discrimination tasks were contrasted with the control task, form discrimination resulted in activations in both occipito temporal and occipito-parietal visual cortical regions, as well as in the right middle-frontal gyrus. Position discrimination resulted in activation in occipito parietal visual cortical regions, the left frontal eye field and the left middle frontal gyrus. These findings are consistent with the view that the processing of visual position information activates occipito-parietal visual regions. On the other hand, the processing of 2D visual form information, in addition to the activation of occipito-temporal neuronal populations, also involves the parietal cortex. Form and position discrimination activated different nonsymmetrical prefrontal fields. Although the visual stimuli were identical, the network of activated cortical fields depended on whether the task was a form discrimination task or a position discrimination task, indicating a strong task dependence of cortical networks underlying form and position discrimination in the human brain. In contrast to former studies, however, these task-dependent macronetworks are overlapping in the posterior parietal cortex, but differentially engage the occipito-temporal and the prefrontal cortex. PMID- 11061338 TI - Analysis of a distributed neural system involved in spatial information, novelty, and memory processing. AB - Perceiving a complex visual scene and encoding it into memory involves a hierarchical distributed network of brain regions, most notably the hippocampus (HIPP), parahippocampal gyrus (PHG), lingual gyrus (LNG), and inferior frontal gyrus (IFG). Lesion and imaging studies in humans have suggested that these regions are involved in spatial information processing as well as novelty and memory encoding; however, the relative contributions of these regions of interest (ROIs) are poorly understood. This study investigated regional dissociations in spatial information and novelty processing in the context of memory encoding using a 2 x 2 factorial design with factors Novelty (novel vs. repeated) and Stimulus (viewing scenes with rich vs. poor spatial information). Greater activation was observed in the right than left hemisphere; however, hemispheric effects did not differ across regions, novelty, or stimulus type. Significant novelty effects were observed in all four regions. A significant ROI x Stimulus interaction was observed - spatial information processing effects were largest effects in the LNG, significant in the PHG and HIPP and nonsignificant in the IFG. Novelty processing was stimulus dependent in the LNG and stimulus independent in the PHG, HIPP, and IFG. Analysis of the profile of Novelty x Stimulus interaction across ROIs provided evidence for a hierarchical independence in novelty processing characterized by increased dissociation from spatial information processing. Despite these differences in spatial information processing, memory performance for novel scenes with rich and poor spatial information was not significantly different. Memory performance was inversely correlated with right IFG activation, suggesting the involvement of this region in strategically flawed encoding effort. Stepwise regression analysis revealed that memory encoding accounted for only a small fraction of the variance (< 16%) in medial temporal lobe activation. The implications of these results for spatial information, novelty, and memory processing in each stage of the distributed network are discussed. PMID- 11061339 TI - Molecular polarity in endothelial cells and tumor-induced angiogenesis. AB - Endothelial cells expose receptors for vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) at the abluminal, basal surface that work as basic regulators of tumor-induced angiogenesis. Their specific localization makes them susceptible to the activity of tumor-released stimulatory factors, like VEGF/VPF, which induce proliferation of the endothelial cell toward the extracellular matrix. At the same time, VEGF/VPF stimulates endothelial cells to expose tissue factor (TF), the high-affinity transmembrane receptor and cofactor for cellular initiation of the plasma coagulation protease cascades through the extrinsic pathway, so generating thrombin. Thrombin exerts a number of activities: it forms an extracellular fibrin barrier from the VEGF/VPF dependent fibrinogen extravasation; it activates progelatinase-A (pro-MMP-2), which destroys the basal membrane, allowing proliferation of endothelial cells (ECs) in the novel tumoral fibrin matrix; finally, it induces EC proliferation, potentiating the VEGF effect. Another important factor exposed at the abluminal endothelial cell surface is membrane type 1 matrix metalloproteinase (MT1-MMP), a membrane-bound metalloproteinase, which also activates progelatinase-A, allowing an alternative pathway to that of thrombin to destroy the basal membrane. In addition, we will see that MT1-MMP is also engaged in a direct, cell-associated fibrinolytic activity, essential for tubulogenesis of the novel outsprouting capillary. PMID- 11061340 TI - Mutational alterations of the p16CDKN2A tumor suppressor gene have low incidence in mesenchymal chondrosarcoma. AB - Mutational inactivation of the cyclin-dependent kinase inhibitors (CDKIs) (p16INK4A/MTS1) tumor suppressor gene has been found in a variety of human tumor types. To investigate the involvement of CDKI abnormality in mesenchymal chondrosarcoma, alterations of CDKIs were examined in human mesenchymal chondrosarcoma tissues using a quantitative DNA/PCR, PCR-SSCP. Seven of 33 specimens (21.2%) showed abnormally low levels of p16CDKN2A amplification, suggesting that the allelic deletion of the gene might be a less frequent event in progression of this tumor. To detect subtle sequence alterations such as point mutations, SSCP analysis of the entire coding region of the p16CDKN2A gene, exons 1, 2, and 3 regions, showed no altered SSCP patterns in 33 mesenchymal chondrosarcoma specimens. A low incidence of genetic alterations of the p16CDKN2A was found in mesenchymal chondrosarcoma. Through this study, we conclude that alteration of the p16CDKN2A gene does not participate significantly in the tumorigenesis of mesenchymal chondrosarcoma. PMID- 11061341 TI - Coamplification of 3-hydroxy-3-methylglutaryl coenzyme A reductase genes in methotrexate-resistant human leukemia cell lines. AB - Methotrexate (MTX)-resistant K562 human myelocytic leukemia sublines with 20- and 200-fold amplified dihydrofolate reductase (DHFR) genes localized to homogeneously staining regions (HSRs) on the long arms of chromosomes 5, 6, and 19 were used to examine whether other genes mapping close to the DHFR genes were coamplified. The gene for 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, located on chromosome 5q13.3-14, was coamplified 4-14-fold, corresponding to the levels of resistance exhibited by these cells. Similar observations were made with a MTX-resistant subline of the promyelocytic leukemia cell line, HL-60R, with 200 gene copies of DHFR. These observations indicate a tight linkage of DHFR and HMG-CoA genes on chromosome 5q. PMID- 11061342 TI - CD18/CD54(+CD102), CD2/CD58 pathway-independent killing of lymphokine-activated killer (LAK) cells against glioblastoma cell lines T98G and U373MG. AB - For natural killer cell-mediated cytolysis (NK-lysis) and lymphokine-activated killer cell-mediated cytolysis (LAK-lysis), the co-stimulatory signals of CD18/CD54(+CD102) and CD2/CD58 pathways are essential. However, in this report, we describe a LAK-lysis that does not depend upon these two pathways. The killed cells were glioblastoma cell lines T98G and U373MG. The LAK cells were induced from peripheral blood lymphocytes in the presence of interleukin-2. 1) The T98G and U373MG did not express CD54 or CD102, but expressed CD58. 2) However, when they were pretreated with an anti-CD58 (TS2/9), the LAK-lysis was not blocked. 3) The LAK-lysis was markedly inhibited by pretreating with Concanamycin A and slightly inhibited by treating with antitumor necrosis factor-related apoptosis inducing ligand (anti-TRAIL) antibody. 4) Nineteen percent of the LAK cells adhered to the T98G. The adhered LAK cells killed it. But nonadherent LAK cells could not kill the T98G or U373MG but killed lymphoblastoma cell lines Raji and NALM-6. These findings suggested that this type of the LAK-lysis might not depend upon the CD18/CD54(+CD102) pathway or CD2/CD58 pathway. The effector cells that killed the T98G and U373MG might not always be the same as the effector cells that killed the other cell lines. The LAK cells contain several subsets, and one of the subsets might kill these two target cell lines. PMID- 11061343 TI - Inhibitory effect of liposomal MDP-Lys on lung metastasis of transplantable osteosarcoma in hamster. AB - MDP-Lys (N2-[(N-acetylmuramyl)-L-alanyl-D-isoglutaminyl]-N6-stearoyl-L-lysine), a macrophage activator, is a lipophilic derivative of muramyl dipeptide (MDP). Multilamellar liposome incorporated MDP-Lys was prepared using phosphatidylcholine and phosphatidylserine by conventional film method, and its inhibitory effect on lung metastasis was compared with MDP-Lys as a solution in hamster's osteosarcoma. The lung metastatic rates after transplantation of the tumor to a lower extremity, in which the extremity was amputated 3 weeks later, were 50% and 100% 3 and 7 weeks, respectively, after transplantation. The rates after amputation were reduced by the treatment with MDP-Lys proportionally to the logarithmic MDP-Lys dose, and the rates 7 weeks after transplantation were 55% and 60%, respectively, in the MDP-Lys solution (50 microg/day) and liposomal MDP Lys (20 microg twice/week) groups. Fifty percent of hamsters treated with liposomal MDP-Lys survived for more than 6 months. Considering that hamsters had a lung metastasis rate of 50% before MDP-Lys treatment, liposomal MDP-Lys given at a dose of 20 microg twice/week was effective for inhibiting lung metastasis at a far lower dose of MDP-Lys than that given as a solution (40 microg vs. 350 microg per week). No significant side effect of liposomal MDP-Lys, as evaluated by the comparison of body weight changes among differently treated hamsters, was detected. This greater inhibitory effect of liposomal MDP-Lys can be considered to be due to the longer retention of the liposomal form in the lung. PMID- 11061344 TI - Angiogenic interactions of vascular endothelial growth factor, of thymidine phosphorylase, and of p53 protein expression in locally advanced gastric cancer. AB - The assessment of the angiogenic profile of tumors may become an important tool as a guide for the inclusion of novel drugs and molecular therapies into the standard chemoradiotherapy policy. Several studies have shown the prognostic importance of microvessel density (MVD) and of angiogenic factor expression in operable gastric cancer. In the present study we investigated, with immunohistochemistry the MVD, the expression of vascular endothelial growth factor (VEGF) and of thymidine phosphorylase (TP) expression, as well as the nuclear expression of p53 protein, in a series of patients with locally advanced inoperable gastric cancer. A strong association of VEGF with TP expression was noted (P = 0.005), and tumors coexpressing these factors had a statistically higher MVD (P = 0.0001). Nuclear p53 accumulation was also related to a high MVD (P = 0.004), and this was independent of VEGF or TP expression. Microvessel density showed a bell-shaped association with prognosis; cases with an intermediate MVD exhibit a favorable outcome (P < 0.05). A trend of nuclear TP expression to define a group of patients with poorer prognosis was noted (P = 0.06), while none of the remaining variables showed any significant association. The immunostaining results allowed the grouping of the angiogenic profile in four major categories: 1) highly vascularized tumors with VEGF and/or TP expression (about 36% of cases); 2) highly angiogenic tumors with p53 nuclear accumulation and low VEGF/TP expression (7% of cases); 3) poorly vascularized tumor with low VEGF/TP and negative nuclear p53 staining (32% of cases); 4) poorly vascularized tumors with TP expression (7% of cases). Specific therapies targeting hypoxia, VEGF, or TP expression as well as p53 gene therapy have entered clinical experimentation or are already available for clinical use. Using the suggested markers more than 80% of locally advanced gastric carcinomas can be grouped in different categories according to their angiogenic profile. Such a categorization may be useful for phase III trials on novel therapies targeting the major angiogenesis-related features studied here. PMID- 11061345 TI - Expression of p53 and bcl-2 in clinically localized prostate cancer before and after neo-adjuvant hormonal therapy. AB - The prognostic significance of p53 and bcl-2 expression in prostate carcinoma is currently under investigation. The aim of the present study was to analyze their expression in diagnostic biopsies and in prostatectomies performed after neo adjuvant hormonal therapy to investigate their role in hormone resistance. One hundred and six patients with advanced prostate carcinoma were treated for 3 months with LHRH analogues before radical surgery. The expression of p53 and bcl 2 was analyzed by immunohistochemistry in all cases of prostatectomy and in available biopsies obtained before treatment, and was correlated with clinicopathologic parameters and follow-up. A significant increase in p53 expression was found following hormonal therapy, whereas no changes were observed in the expression of bcl-2. The increase in p53 did not correlate with the presence of therapy-induced morphological changes in prostate cancers, but it did correlate significantly with histologic grade and pathologic stage, biochemical progression of the disease, and short overall survival. At multivariate analysis, only grade and stage proved to be independent predictors of shorter survival. There were no correlations between bcl-2 and clinicopathologic variables whether in biopsies or in prostatectomies. The unfavorable clinical course associated with p53-positive carcinomas suggests that neo-adjuvant hormonal therapy may cause the selection of minor p53 mutated clones, rather than the induction of wild-type p53. In any case, the enhanced expression of p53 could label hormone resistant cancers for further adjuvant therapy. PMID- 11061346 TI - Endothelial dysfunction in the pulmonary vascular bed. AB - The pulmonary endothelium modulates vascular tone by the release of endothelium derived constricting (EDCF) and relaxing (EDRF) factors, among them endothelin-1, nitric oxide, prostacyclin, and putative endothelium-derived hyperpolarizing factors. Abnormalities in EDCF and EDRF generation have been demonstrated in a number of cardiopulmonary disease states, such as primary and secondary pulmonary hypertension, chronic obstructive lung disease, cardiopulmonary bypass, and congestive heart failure. An imbalance between EDCF and EDRF, termed "pulmonary endothelial dysfunction," may contribute to the alteration in vascular tone characteristic of pulmonary disease. The following review summarizes the present knowledge of the role of EDCF and EDRF in such processes with major focus on pulmonary endothelial dysfunction in hypoxia-induced pulmonary hypertension. PMID- 11061347 TI - Biochemical and metabolic effects of very-low-salt diets. AB - In cross-cultural studies, very low sodium intakes are associated with a low prevalence of hypertension and minimal increase of blood pressure with aging. Disorders of lipid and carbohydrate metabolism are rare. In short-term clinical studies, very low sodium intake (<50 mmol/d) has been associated with greater values for total and low-density lipoprotein cholesterol, fasting and post glucose insulin, uric acid, plasminogen activator inhibitor-1, and activity of the renin-angiotensin system. Thus, the long-term safety of the very-low-salt diets suggested by these observations, in which sodium is one of many differences between population groups, is not entirely consonant with the short-term clinical trials data in which sodium is studied as an isolated intervention. This may reflect transient effects of abrupt and large changes in sodium consumption. Nevertheless, differences in diet composition and nutrient intake other than sodium including potassium, magnesium, and a range of antioxidants may also contribute to the discrepancies between ecological observations and clinical studies. Further research on the effects of selective changes of dietary sodium versus more global changes in diet composition on biochemical and hemodynamic variables could provide the basis for an even more effective public health policy. PMID- 11061348 TI - Dyspnea scales in the assessment of illiterate patients with chronic obstructive pulmonary disease. AB - BACKGROUND: Multiple physiological, psychological, social and environmental factors may affect the perception of dyspnea. Although different scales have been used to record the severity of dyspnea in subjects with chronic obstructive pulmonary disease (COPD), none has reported evaluating the properties of such tools in illiterate patients. The objective of this study was to evaluate the reliability and features of concurrent validity of 4 dyspnea scales in illiterate (IL) subjects with COPD. METHODS: One hundred COPD patients submitted to spirometry and were asked to score their breathlessness using a visual analogue scale (VAS), a numerical rating scale (NRS), the Borg scale (BS), and the basal dyspnea index (BDI). Each scale was presented to the patients before and after they had performed spirometry and measurement of residual volume. The obtained scores were analyzed according to the literacy status of the patients. RESULTS: Thirty-three patients were classified as IL and 67 as literate (L). Both groups showed similar respiratory impairment and median scores of dyspnea (VAS, L = 45.0, IL = 49.0; NRS, L = 5.0, IL = 5.0; BS, L = 3.0, IL = 3.0; BDI, L = 5.0, IL = 4.0). No significant differences were found between the dyspnea scores obtained before and after spirometry for all scales in both groups. The degree of correlation between forced expiratory volume in 1 second (FEV1) and usual dyspnea evaluated by BDI did not show a statistical difference between the two groups (L, r = 0.37; IL, r = 0.51). CONCLUSION: The employed dyspnea scales showed comparable reliability in both L and IL COPD subjects. PMID- 11061349 TI - Myocardial function during chronic food restriction in isolated hypertrophied cardiac muscle. AB - BACKGROUND: The effect of food restriction (FR) on myocardial performance has been studied in normal hearts. Few experiments analyzed the effects of undernutrition on hearts subjected to cardiac overload. The aim of this study was to determine whether chronic FR promotes more significant changes in hypertrophied hearts than in normal hearts. METHODS: Myocardial performance was studied in isolated left ventricular papillary muscle from young male spontaneously hypertensive rats (SHR) and age-matched normotensive Wistar-Kyoto rats (WKY) submitted to FR or to control diet. The animals subjected to FR were fed 50% of the amount of food consumed by control groups for 60 days. Isolated muscles were studied while contracting isometrically and isotonically. RESULTS: FR decreased the body weight and the left ventricular weight in both groups. FR increased the left ventricular weight-to-body weight ratio in the WKY rats and tended to decrease this ratio in SHR (P = 0.055). The arterial systolic pressure was greater in SHR than in WKY groups and did not change with FR. In the animals with normal diet, myocardial performance was better in SHR than in WKY. FR increased time to tension to fall from peak to 50% of peak tension and time to peak tension in the WKY rats and time to peak tension in the SHR. CONCLUSIONS: FR for 60 days has a trend to attenuate the development of cardiac hypertrophy and does not promote more mechanical functional changes in the hypertrophied myocardium than in the normal cardiac muscle. PMID- 11061350 TI - Indoor air effects after building renovation and in manufactured homes. AB - PROBLEM: The objective was to measure and compare the neurobehavioral and respiratory effects of exposures to indoor air in people living in manufactured homes and occupying buildings during renovation and compare them with effects on subjects exposed to formaldehyde at work. METHODS: Ten people living in manufactured homes and 10 people exposed to chemicals during renovation of their offices or homes had measurements made of balance, visual fields, reaction time, hearing, grip strength, and vibration sense. Psychological measurements included cognition, recall, perceptual motor speed, long-term memory, and mood states. RESULTS: Exposures to indoor air were associated with abnormal simple and choice reaction time, abnormal balance with the eyes open and with the eyes closed, abnormalities of color confusion index, scotoma in visual fields, reduced verbal recall, digit symbol score, and elevated abnormal moods. The effects on the two groups of 10 were similar and resembled those from formaldehyde exposure but with less cognitive impairment. CONCLUSIONS: Adverse effects from indoor air in manufactured homes and during renovations were less severe but similar to those from occupational formaldehyde exposures. This suggests that formaldehyde has a major role in health problems from indoor air. PMID- 11061351 TI - Native valve infective endocarditis: what is the optimal timing for surgery? AB - IE remains a dreaded disease masquerading under a myriad of presentations in an evolving epidemiological environment. In our continuing endeavor against this deadly disease, echocardiography has evolved into an indispensable diagnostic tool to define structural complications and guide therapy. Timing of surgical intervention for IE remains a subject of intense debate and depends on the cardiac and systemic complications of the infection, the virulence of the organism, and the responsiveness to medical therapy. A judicious agreement among cardiologist, cardiovascular surgeon, and infectious disease specialist should define whether surgical intervention is warranted and, if so, the optimal timing. Further optimization of guidelines will help in the diagnosis and treatment of endocarditis but will never be a substitute for sound judgment and experience. PMID- 11061352 TI - In pursuit of the prevention of breast cancer. PMID- 11061353 TI - Post-herpes encephalitic anterior pituitary insufficiency with hypothermia and hypotension. AB - A 49-year-old man with herpes simplex encephalitis at age 22 was admitted with hypotension (90/60 mm Hg) and hypothermia (33.7 degrees C). His blood pressure was 80-90/50-60 mm Hg, with temperatures averaging 35 degrees C, for at least 3 years before admission. Evaluation of his hypothermia and hypotension revealed a low free triiodothyronine, low normal thyrotropin, luteinizing hormone < 2 mIU/L, follicle stimulating hormone <3 mIU/L, and low testosterone of 1.39 ng/dL. A baseline cortisol of 13.9 microg/dL was stimulated to 41.8 microg/dL with corticotropin, indicating he had partial anterior hypopituitarism with an intact pituitary-adrenal axis. Posterior pituitary function was normal. MRI revealed a "bright" posterior pituitary on a T1-weighted image, further indicating a normal posterior pituitary. Extensive decreased T1-weighting on MRI in the right and left temporal lobes was consistent with encephalomalacia. With thyroid hormone replacement, his blood pressure increased to 110/70 mm Hg with a temperature of 37 degrees C. PMID- 11061354 TI - Unilateral internuclear ophthalmoplegia and recovery with thiamine in Wernicke syndrome. AB - Internuclear ophthalmoplegia is usually caused by multiple sclerosis, tumors, or vascular lesions of the brain stem. We report a patient with Wernicke syndrome who presented with a right-sided internuclear ophthalmoplegia. He recovered completely with intravenous thiamine (vitamin B1). There were no lesions in the magnetic resonance image (MRI) of the brain, suggesting a derangement at the cellular level as the cause. PMID- 11061355 TI - Rapid growth of a basilar aneurysm. AB - We describe a patient who developed, over a 22-month period, a giant aneurysm of his basilar artery. A prior MRI of the brain done for nonspecific symptoms showed a normal brainstem and basilar artery. At presentation, he had a repeat MRI scan for a 4-month history of a partial right oculomotor nerve palsy and left hemiparesis. The MRI revealed a giant aneurysm of the top of the basilar artery. This was treated by angiographic placement of Guglielmi detachable coils (GDC) after surgical intervention was deemed unfeasible. This case illustrates the acquired nature of intracranial aneurysms. All inoperable intracranial aneurysms should be closely monitored and MRI and MR angiography may currently be the best noninvasive methods for this purpose. Intra-arterial GDC embolization of aneurysms is an alternative treatment when surgery is not possible. PMID- 11061356 TI - Left hydronephrosis caused by Crohn disease successfully treated conservatively. AB - We report the case of a 35-year-old man who presented with fever, diarrhea, and a left abdominal mass. Diagnostic studies confirmed Crohn disease and revealed an abdominal mass obstructing the left ureter with hydroureter and hydronephrosis. The patient was successfully treated conservatively, with corticosteroids and mesalamine, A review of the literature indicates a predominance of right ureteral involvement in Crohn disease, associated with a high incidence of ileocecal disease. Most of these patients were treated surgically, with resection of ileocecal lesion and/or ureterolysis. Ureteral obstruction as a complication of Crohn disease is discussed, with emphasis on conservative treatment. PMID- 11061357 TI - Amiodarone-induced thyrotoxicosis type 2: a case report and review of the literature. AB - Amiodarone-induced thyrotoxicosis (AIT) is not an uncommon complication in countries with low iodine intake. Two types of AIT have been described. Type 1 is associated with an underlying disorder of the gland that becomes clinically evident because of the high intrathyroidal iodine content, which acts as a trigger for hyperthyroidism. Type 2 is characterized by an ongoing inflammatory process of the thyroid, with derangement of the parenchyma causing the release of thyroid hormones into the circulation. Yet, there are no definitive tools for the differential diagnosis of these two entities, and the therapeutic approach is still a subject of controversy. A case of AIT type 2 successfully treated with steroids is described followed by a review of the literature. PMID- 11061358 TI - The heterogeneity of Castleman disease: report of five cases and review of the literature. AB - Castleman disease (CD; angiofollicular lymphoid hyperplasia) is a heterogeneous group of lymphoproliferative disorders of uncertain cause. Three histologic variants (hyaline vascular, plasma cell, and mixed) and two clinical types (localized and multicentric) of CD have been described. We report 5 cases of CD treated in our institute and review the literature about the management of this relatively rare disorder. Localized and multicentric CD may be different clinical disorders with overlapping histologic features. Localized disease generally presented with a single enlarged lymph node or widening of the mediastinum, whereas multicentric disease is a systemic lymphoproliferative disorder characterized by lymphadenopathy, hepatosplenomegaly, constitutional symptoms, anemia, hypoalbuminemia, and hypergammaglobulinemia. Unlike the localized type, for which surgical excision is curative regardless of the histologic type, multicentric disease often necessitates aggressive systemic therapy and portends a poorer outcome. PMID- 11061359 TI - Thrombocytopenia associated with octreotide. AB - A 42-year-old woman with a history of hepatitis C-induced cirrhosis, gastrointestinal bleeding, and alcohol abuse presented to the hospital with hematemesis and melena. Based on our previous experience, octreotide (Sandostatin) therapy was started at 50 mg/hr and continued for 5 days. Platelet count on admission (122 x 10(9)/L) dropped immediately after octreotide therapy was started; upon discontinuation, platelet count began trending up from 72 x 10(9)/L. However, octreotide was not suspected at this point as the cause of thrombocytopenia. In a subsequent admission, octreotide was again administered with a resultant prompt decrease in platelet count. To our knowledge, this is only the second case report of octreotide-induced thrombocytopenia, and the first case of this adverse effect demonstrated by inadvertent rechallenge. PMID- 11061360 TI - Exercise training as a form of cardiovascular therapy in chronic heart failure. PMID- 11061361 TI - Endothelial dysfunction in chronic heart failure: some new basic mechanisms. AB - Endothelial dysfunction contributes to the maintenance of peripheral vasoconstriction and abnormal vascular compliance in chronic heart failure. Endothelial dysfunction results in an imbalance between vasodilation and vasoconstriction, particularly when adjustments in blood flow are required. Recently, new factors have been recognized to determine endothelial dysfunction: a) disturbances of the L-arginine/nitric oxide pathway, either at the enzymatic or substrate level; b) increased synthesis of endothelin-1; c) microvessel structural remodeling; d) increased adhesive properties to blood cell components; and e) apoptotic cell injury. The understanding of the complex interplay among these factors is the basis for development of new targeted strategies to correct endothelial dysfunction in chronic heart failure. PMID- 11061362 TI - Pharmacogenetics: a molecular sophistication or a new clinical tool for cardiologists? AB - The study of genetic risk factors for multifactorial diseases is attracting increasing interest. In particular interest has been focused on the interaction between genetic polymorphisms and environmental factors in determining the risk of disease. Among environmental factors therapeutic approaches should be considered. Therapeutic responses to a given drug, failure of drug efficacy, interindividual variability in side effects and toxicity of drugs could be at least partially accounted for by genetic polymorphisms. This paper summarizes the presently available applications of genetic concepts to some drugs commonly used in patients with cardiovascular disease. Statins and probucol fail to lower cholesterol levels in carriers of specific polymorphisms. The progression of cardiovascular disease is decreased by pravastatin only when certain polymorphisms are present. Induction problems and bleeding complications of warfarin occur in subgroups of populations carrying specific genetic variants of key enzymes in the drug metabolism. A new interpretation of the results of a thrombosis prevention trial will be given in the light of a genetic approach to pharmacology; indeed, prevention and treatment of thrombotic disease could be better focused on the basis of this knowledge. Future clinical trials and cost effectiveness evaluation of drugs should be conducted taking these gene-drug interactions into account. PMID- 11061363 TI - Your cardiac patient wants to become a mother. Risk considerations and advice. Part II--Your cardiac patient is pregnant. AB - In this part the most updated indications for evidence-based counseling of cardiac patients during pregnancy, labor and the postpartum period are reviewed. Moreover, indications and contraindications for the use of some cardiac drugs during pregnancy and lactation are provided. PMID- 11061364 TI - Long-term changes in the response of conductance and resistance coronary vessels to endothelium-dependent and independent vasodilators. A double-blind placebo controlled study of the effect of a 6-month treatment with cilazapril. AB - BACKGROUND: The aim of this study was to assess the effect of a 6-month treatment with the inhibitor of the angiotensin-converting enzyme cilazapril on the response of conductance and resistance vessels to endothelium-dependent and independent vasodilators, in a randomized placebo-controlled parallel group single center study. METHODS: Quantitative angiographic and Doppler flow time averaged peak velocity measurements were performed in an artery with < 30% diameter stenosis after sequential selective intracoronary injection of papaverine (7 mg), acetylcholine (0.036, 0.36 and 3.6 microg/ml at 2 ml/min) and isosorbide dinitrate. Repeated assessment was performed after a 6-month treatment with cilazapril 20 mg/day or placebo. Thirty-four patients were enrolled in the study undergoing elective percutaneous coronary interventions for stable angina. Main outcome measures were percent differences from baseline and absolute measurements of mean coronary cross-sectional area, coronary flow time-averaged peak velocity and flow resistance in the initial study and at follow-up for the placebo and the treated group. RESULTS: No significant differences between the placebo and the treated group were observed in the modifications of cross sectional area after acetylcholine and isosorbide dinitrate and in the response of time-averaged peak velocity to papaverine. After the maximal concentration of acetylcholine a high but statistically not significant increase in flow and a decrease in flow resistance were observed in the treated group (medians: 45% increase vs 4% increase for coronary flow, and 44% decrease vs 1% increase for flow resistance in the cilazapril and in the treated group, respectively, p = NS). CONCLUSIONS: In patients with coronary artery disease, a 6-month treatment with 20 mg of cilazapril/day did not modify the response to endothelium independent and dependent vasodilators of epicardial arteries without any significant stenoses but induced a consistent, although not significant, increase in flow and decrease in flow resistance after acetylcholine. PMID- 11061365 TI - Non-invasive estimation of right atrial pressure by combined Doppler echocardiographic measurements of the inferior vena cava in patients with congestive heart failure. AB - BACKGROUND: In patients with congestive heart failure, evaluation of right atrial pressure (RAP) provides useful therapeutic, functional and prognostic information. The aim of this study was to investigate whether a combination of inferior vena cava variables measured by Doppler echocardiography could provide a reliable non-invasive estimate of RAP. METHODS: One hundred consecutive patients with severe congestive heart failure (ejection fraction 24 +/- 6%) due to dilated cardiomyopathy were evaluated by simultaneous Doppler echocardiography and hemodynamic studies. RAP, end-expiratory (IVCDmax) and end-inspiratory (IVCDmin) diameters of the inferior vena cava, its collapse index [CIIVC = (IVCDmax - IVCDmin/IVCDmax)*100] and systolic fraction of forward inferior vena cava flow were measured and correlated by both single and multilinear regression analysis. The accuracy of generated equations was tested in a separate testing group of 61 patients at baseline and a subgroup of 20 patients after loading manipulations, prospectively studied in the same methodological setting. RESULTS: All Doppler echocardiographic variables were correlated with RAP. The IVCDmin showed the strongest correlation (r = 0.84, p < 0.0001). Stepwise regression analysis identified two equations for predicting RAP: 1) RAP = (6.4*IVCDmin + 0.04*CIIVC - 2) (r = 0.82, p < 0.0001, SEE 1.7 mmHg) in all patients, and 2) RAP = (4.9*IVCDmin + 0.01*CIIVC - 0.2) (r = 0.92, p < 0.0001, SEE 1.2 mmHg) in patients without tricuspid regurgitation. In the testing group estimated and measured RAP was strongly correlated at baseline (r = 0.95, SEE 1.3 mmHg, p < 0.00001) and after loading manipulations (r = 0.96, SEE 1.2 mmHg, p < 0.00001). The agreement between invasive and non-invasive measurements of RAP in identifying patients with normal (< or = 5 mmHg), moderately increased (< 5 RAP < 10 mmHg) and markedly increased (> or = 10 mmHg) RAP was 81 or 93% using equation 1 or 2, respectively. CONCLUSIONS: Our results provide evidence that in patients with congestive heart failure indices derived from Doppler measurements of the inferior vena cava can be used to produce an accurate, strong and non-invasive estimate of RAP. This is another example of the usefulness of Doppler echocardiography in evaluating hemodynamic profile and its changes in patients with congestive heart failure. Echocardiographic assessment of the inferior vena cava should be included in the evaluation of patients with congestive heart failure. PMID- 11061366 TI - Post-discharge recurrences of new-onset atrial fibrillation following cardiac surgery: impact of low-dose amiodarone and beta-blocker prophylaxis. AB - BACKGROUND: Atrial fibrillation (AF) is the most frequent complication following cardiac surgery. It occurs mostly between the second and fourth postoperative days and often recurs within 30 days after surgery. While postoperative AF has been well analyzed, post-discharge recurrences of AF have not been clearly described even if they are reported as a frequent cause of re-hospitalization. METHODS: We followed up 185 patients for 10 +/- 5 months with the aim of characterizing the post-discharge recurrences of AF. All patients had recently undergone cardiac surgery complicated by AF and were in sinus rhythm at the time of admission to our Center. We also compared the efficacy of the main prophylactic regimens adopted in the referral Centers (amiodarone, beta-blockers, amiodarone plus beta-blockers) during the first postoperative month. RESULTS: In the first postoperative month AF recurred after discharge in 60 patients. The event rate was not different in patients treated with amiodarone and controls (47 vs 50%, p = NS), while it was significantly lower in patients taking beta blockers either alone or associated with amiodarone (10 and 9% respectively, p = 0.002). At the end of follow up (10 +/- 5 months), AF persisted in 3 out of 176 study patients (1.7%). CONCLUSIONS: In patients undergoing cardiac surgery, post discharge recurrences of AF are frequent during the first postoperative month and have a clinical relevance. Beta-blockers (not amiodarone) seem to be an effective prophylactic measure. The phenomenon tends to vanish in the long term, and a chronic prophylaxis is not justified. PMID- 11061367 TI - Totally endoscopic atrial septal defect closure using robotic techniques: report of two cases. AB - BACKGROUND: The development of minimally invasive cardiac surgery has shown good clinical results with shorter recovery time and better cosmetic results. We report 2 cases of totally endoscopic atrial septal defect (ASD) closure using a robotic system. Open-heart closure of an ASD without opening the chest has never been previously reported. METHODS: Following percutaneous cannulation for cardiopulmonary bypass, aortic occlusion and delivery of cardioplegia, 2 patients with an ASD were successfully operated on using a robotic surgical device. After exclusion of the right lung, two robotic arms and an endoscopic camera were inserted through ports in the right hemithorax. A fourth port was inserted for an accessory endoscopic instrument. The ASD closure was carried out with interrupted stitches in one case and with a continuous suture in the other. RESULTS: Cardiopulmonary bypass and cardioplegic arrest times were respectively 130 and 75 min in the first and 87 and 60 min in the second case. Extubation was carried out 3 and 5 hours postoperatively. Both patients resumed a totally normal lifestyle 1 week after the operation. CONCLUSIONS: Totally endoscopic open-heart ASD closure can be carried out safely using robotic techniques with rapid postoperative recovery and excellent cosmetic results. This modality of treatment can be considered an alternative to the transcatheter closure of ASD. PMID- 11061368 TI - Cardiogenic shock due to acute left main coronary artery occlusion: successful treatment with primary angioplasty. AB - A 56-year-old patient was admitted with cardiogenic shock due to an acute anterior myocardial infarction. Cardiac catheterization with coronary angiography disclosed a thrombotic occlusion of the left main coronary artery. Prompt mechanical recanalization of the infarct-related artery with multiple stent implantations associated with prolonged circulatory and respiratory supports allowed for a partial recovery of the left ventricular function and the discharge of the patient. PMID- 11061369 TI - Ebstein's anomaly associated with ventricular septal defect and pulmonary stenosis. PMID- 11061370 TI - Recent developments regarding heart transplantation, in particular the difficult donor harvesting. PMID- 11061371 TI - A message from the Executive Vice President regarding the ECFVG program. Educational Commission for Foreign Veterinary Graduates. PMID- 11061372 TI - In praise of women who choose career and family. PMID- 11061373 TI - In opposition of forced molting. PMID- 11061374 TI - Concerns about euthanasia report. PMID- 11061375 TI - Feels alternative and complementary therapies need scientific evidence. PMID- 11061376 TI - Contamination of feedstuffs caused by farm dogs. PMID- 11061377 TI - Responding to blame by blamectomy and blamotomy. PMID- 11061378 TI - What is your diagnosis? Diffuse increase in pulmonary opacity with a bronchial pattern and bronchiectasis. PMID- 11061379 TI - Predictors of owner response to companion animal death in 177 clients from 14 practices in Ontario. AB - OBJECTIVE: To identify predictors of grief and client desires and needs as they relate to pet death. DESIGN: Cross-sectional mail survey. SAMPLE POPULATION: 177 clients, from 14 randomly selected veterinary practices, whose cat or dog died between 6 and 43 days prior to returning the completed questionnaire. PROCEDURE: Veterinary practices were contacted weekly to obtain the names of clients whose pets had died until approximately 200 clients were identified. Clients were contacted by telephone, and a questionnaire designed to measure grief associated with pet death was mailed to those willing to participate within 1 to 14 days of their pet's death. The questionnaire measured potential correlates and modifiers of grief and included three outcome measures: social/emotional and physical consequences, thought processes, and despair. Demographic data were also collected. RESULTS: Approximately 30% of participants experienced severe grief. The most prominent risk factors for grief included level of attachment, euthanasia, societal attitudes toward pet death, and professional support from the veterinary team. CONCLUSIONS AND CLINICAL RELEVANCE: Bivariate and multivariate analyses highlighted the impact owners' attitudes about euthanasia and professional intervention by the veterinary team had on reactions to pet death. Owners' perceptions of societal attitudes, also a predictor of grief, indicate that grief for pets is different than grief associated with other losses. PMID- 11061380 TI - Racing boards and the practice of veterinary medicine. PMID- 11061381 TI - Implementation and assessment of a career and life skills program for matriculating veterinary medical students. PMID- 11061382 TI - Survey of chief livestock officials regarding bioterrorism preparedness in the United States. PMID- 11061383 TI - Controversies and clarifications regarding bovine lentivirus infections. Subcommittee for the Bovine Retrovirus Committee, US Animal Health Association. PMID- 11061384 TI - Results of ambulatory electrocardiography in overtly healthy Doberman Pinschers with echocardiographic abnormalities. AB - OBJECTIVE: To identify, by means of 24-hour ambulatory electrocardiography, electrocardiographic abnormalities in overtly healthy Doberman Pinschers in which results of echocardiography were abnormal. DESIGN: Clinical case series. ANIMALS: 56 (35 male, 21 female) overtly healthy Doberman Pinschers with echocardiographic evidence of cardiomyopathy on initial examination that subsequently died of cardiomyopathy. PROCEDURE: Twenty-four-hour ambulatory electrocardiographic (Holter) recordings obtained at the time of initial examination were reviewed. For all dogs, scan quality was > 90%. RESULTS: Initial Holter recordings of all 56 dogs contained ventricular premature contractions (VPC). Thirty-six (65%) dogs had > 1,000 VPC/24 h, 17 (31%) had > 5,000 VPC/24 h, and 11 (19%) had > 10,000 VPC/24 h. Fifty-four (96%) dogs had couplets of VPC, 37 (66%) had triplets of VPC, and 36 (64%) had episodes of nonsustained (< 30 seconds) ventricular tachycardia. Number of VPC/24 h during the initial Holter recordings was positively correlated with numbers of couplets and triplets of VPC and number of ventricular escape beats and negatively correlated with left ventricular fractional shortening. Twenty-eight dogs died suddenly prior to the putative onset of congestive heart failure. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that along with echocardiography, 24-hour ambulatory electrocardiography can be used to help identify overtly healthy Doberman Pinschers with cardiomyopathy. PMID- 11061385 TI - Video-assisted thoracoscopic division of the ligamentum arteriosum in two dogs with persistent right aortic arch. AB - Video-assisted division of the ligamentum arteriosum can be performed successfully in dogs with minimal postoperative complications and hospitalization time. Single-lung ventilation and thoracic insufflation are not mandatory and standard instrumentation may be used for most of the procedure. PMID- 11061386 TI - Complications and outcomes associated with use of gastrostomy tubes for nutritional management of dogs with renal failure: 56 cases (1994-1999). AB - OBJECTIVE: To evaluate complications and outcomes associated with use of gastrostomy tubes in dogs with renal failure. DESIGN: Retrospective study. ANIMALS: 56 dogs. PROCEDURE: Medical records were reviewed for dogs with renal failure that were treated by use of gastrostomy tubes. RESULTS: Mean +/- SD BUN concentration was 134 +/- 79 mg/dl and mean serum creatinine concentration was 9.0 +/- 3.8 mg/dl. Low-profile gastrostomy tubes were used for initial placement in 10 dogs, and traditional gastrostomy tubes were used in 46 dogs. Mild stoma site complications included discharge, swelling, erythema, and signs of pain in 26 (46%) of dogs. Twenty-six gastrostomy tubes were replaced in 15 dogs; 11 were replaced because of patient removal, 6 were replaced because of tube wear, and 3 were replaced for other reasons. Six tubes were replaced by low-profile gastrostomy tubes. Gastrostomy tubes were used for 65 +/- 91 days (range, 1 to 438 days). Eight dogs gained weight, 11 did not change weight, and 17 lost weight; information was not available for 20 dogs. Three dogs were euthanatized because they removed their gastrostomy tubes, 2 were euthanatized because of evidence of tube migration, and 1 died of peritonitis. CONCLUSIONS AND CLINICAL RELEVANCE: Gastrostomy tubes appear to be safe and effective for improving nutritional status of dogs with renal failure. PMID- 11061387 TI - Palmar-plantar axial sesamoidean approach to the digital flexor tendon sheath in horses. PMID- 11061388 TI - Clinical application of a polymerase chain reaction assay in the diagnosis of pneumonia caused by Rhodococcus equi in a horse. AB - Diagnosis of pneumonia caused by Rhodococcus equi can be made more rapidly by use of a polymerase chain reaction (PCR) assay than by use of conventional bacteriologic culture techniques. Use of a PCR assay aids in the differentiation between virulent and avirulent strains of R equi, and the assay may be used to identify R equi in feces and soil of breeding farms. PMID- 11061389 TI - Removal of large fragments of the extensor process of the distal phalanx via arthrotomy in horses: 14 cases (1992-1998). AB - OBJECTIVE: To evaluate the outcome of horses with large fragments of the extensor process of the distal phalanx that were removed by use of arthrotomy. DESIGN: Retrospective study. ANIMALS: 14 horses with large fragments of the extensor process of the distal phalanx. PROCEDURE: Medical records for horses with large fragments of the extensor process that were removed by use of arthrotomy were reviewed. Data retrieved from medical records included signalment, use of horse, affected limb, lameness history, lameness examination findings, radiographic findings, surgical technique, and outcome. Follow-up evaluation was obtained by telephone interview. RESULTS: Most affected horses were < 5 years old and had a history of chronic lameness. Lameness grade ranged from 1/5 to 4/5. Fragments involved 20 to 45% of the dorsopalmar articular surface of the distal phalanx. Eight of 14 horses had a successful outcome. Outcome was not associated with age, duration or severity of lameness, or fragment size. CONCLUSIONS AND CLINICAL RELEVANCE: Despite involvement of a large portion of the articular surface and use of arthrotomy, joint instability and permanent soft tissue injury was not a problem in most horses. Outcome may be improved by selection of horses with lameness of < 2 years' duration and careful management after surgery. A fair prognosis may be anticipated for removal of large fragments of the extensor process via arthrotomy. PMID- 11061390 TI - Association of serologic status for Neospora caninum with postweaning weight gain and carcass measurements in beef calves. AB - OBJECTIVE: To determine the seroprevalence for Neospora caninum in a population of beef calves in a feedlot and the association of serologic status with postweaning weight gain and carcass measurements. DESIGN: Longitudinal observational study. ANIMALS: 1,009 weaned beef steers from 92 herds. PROCEDURE: Samples were obtained from all steers at time of arrival at a feedlot. Serologic status for Neospora spp was determined, using an agglutination test. Results of serologic testing were compared with calf growth and carcass data, using multivariate regression with generalized estimating equations. RESULTS: Of 1,009 calves, 131 (12.98%) were seropositive, and 54 of 92 (58.7%) consignments had > or = 1 seropositive calf. Median within-consignment prevalence for consignments in which there was > or = 1 seropositive calf was 20%. Seropositive status was associated with significant reductions in average daily gain, live body weight at slaughter, and hot carcass weight and an increase in ribeye area-to-hot carcass weight ratio. Seropositive status also was associated with significant increases in cost of treatment and significant reductions in income. Sick seropositive calves had the highest cost of treatment. An economic loss of $15.62/calf was projected for seropositive calves. CONCLUSIONS AND CLINICAL RELEVANCE: Significant reductions in postweaning weight gain, carcass weight, and economic return were associated with detection of antibodies to N caninum in beef calves in a feedlot. PMID- 11061391 TI - Spatial associations among density of cattle, abundance of wild canids, and seroprevalence to Neospora caninum in a population of beef calves. AB - OBJECTIVE: To determine the epidemiologic plausibility of a sylvatic transmission cycle for Neospora caninum between wild canids and beef cattle. DESIGN: Spatial analysis study. ANIMALS: 1,009 weaned beef steers from 94 beef herds in Texas. PROCEDURE: Calves were grouped on the basis of seroprevalence for N caninum and ecologic region in Texas. The Morans I test was used to evaluate spatial interdependence for adjusted seroprevalence by ecologic region. Cattle density (Number of cattle/259 km2 [Number of cattle/100 mile2] of each ecologic region) and abundance indices for gray foxes and coyotes (Number of animals/161 spotlight transect [census] km [Number of animals/100 census miles] of each ecologic region) were used as covariates in spatial regression models, with adjusted seroprevalence as the outcome variable. A geographic information system (GIS) that used similar covariate information for each county was used to validate spatial regression models. Results-Spatial interdependence was not detected for ecologic regions. Three spatial regression models were tested. Each model contained a variable for cattle density for the ecologic regions. Results for the 3 models revealed that seroprevalence was associated with cattle density and abundances of gray foxes, coyotes, or both. Abundances of gray foxes and coyotes were collinear. Results of a GIS-generated model validated these spatial models. CONCLUSIONS AND CLINICAL RELEVANCE: In Texas, beef cattle are at increased risk of exposure to N caninum as a result of the abundance of wild canids and the density of beef cattle. It is plausible that a sylvatic transmission cycle for neosporosis exists. PMID- 11061392 TI - Cardiorespiratory effects of four alpha2-adrenoceptor agonist-ketamine combinations in captive red wolves. AB - OBJECTIVE: To evaluate the cardiopulmonary effects of immobilizing doses of xylazine-ketamine (XK), medetomidine-ketamine (MK), medetomidine-ketamine acepromazine (MKA), and medetomidine-butorphanol-ketamine (MBK) in captive red wolves. DESIGN: Prospective study. ANIMALS: 32 adult captive red wolves. PROCEDURE: Wolves were randomly assigned to 1 of 4 treatment groups: XK, MK, MKA, or MBK. Physiologic variables measured included heart rate, blood pressure, respiratory rate, tidal volume, oxygen-hemoglobin saturation (Spo2), end-tidal CO2, arterial blood gases, and rectal temperature. Induction time, muscle relaxation, and quality of recovery were assessed. RESULTS: Heart rates were lower in wolves in the MBK group than for the other groups. All 4 drug combinations induced considerable hypertension, with diastolic pressures exceeding 116 mm Hg. Blood pressure was lowest in wolves receiving the MBK combination. Respiratory rate was significantly higher in wolves receiving XK, MK, and MKA. Tidal volumes were similar for all groups. Wolves receiving XK, MK, and MKA were well-oxygenated throughout the procedure (SPo2 > 93%), whereas those receiving MBK were moderately hypoxemic (87% < Spo2 < 93%) during the first 20 minutes of the procedure. Hyperthermia was detected initially following induction in all groups. CONCLUSIONS AND CLINICAL RELEVANCE: The alpha2-adrenoceptor agonist-ketamine combinations provide rapid reversible anesthesia for red wolves but cause severe sustained hypertension. Such an adverse effect puts animals at risk for development of cerebral encephalopathy, retinal hemorrhage, pulmonary edema, and myocardial failure. Although the MBK combination offers some advantages over the others, it is advised that further protocol refinements be made to minimize risks associated with acute hypertension. PMID- 11061393 TI - Differences and similarities between behavioral and internal medicine. PMID- 11061394 TI - Coronary revascularisation in the private and public sectors. Does the Australian healthcare system deliver value for money? PMID- 11061395 TI - Evidence-based management of melanoma. Clinical practice guidelines are now available for managing this cancer which occurs in relatively young people and is a particular problem in Australia. PMID- 11061396 TI - Cancer among Indigenous Australians: time for decisive action. Cancers in Indigenous people are largely preventable. PMID- 11061397 TI - Coronary angiography and coronary artery revascularisation rates in public and private hospital patients after acute myocardial infarction. AB - OBJECTIVE: To determine the rates of coronary angiography or coronary artery revascularisation procedures in patients with acute myocardial infarction (AMI) managed in private versus public hospitals. DESIGN: Case record linkage analysis of data from the Victorian Inpatient Minimum Dataset for admissions for AMI in the 12 months after the index admission. SETTING: Victorian acute care hospitals from July 1995 to December 1997. PATIENTS: Victorian residents aged 15-85 years admitted to hospital with AMI. MAIN OUTCOME MEASURES: Rates of coronary angiography or coronary artery revascularisation procedures after AMI. RESULTS: Compared with public patients in public hospitals, patients with AMI managed in private hospitals were more likely to undergo coronary angiography (rate ratio [RR], 2.17; P< 0.001; 95% CI, 2.06-2.29), coronary angioplasty or stenting (RR, 3.05; P<0.001; 95% CI, 2.82-3.31), and coronary artery bypass grafting (RR, 1.95; P<0.001; 95% CI, 1.79-2.14). Once coronary angiography had been performed, patients in private hospitals were more likely to undergo angioplasty or stenting (RR, 1.94; P<0.001; 95% CI, 1.79-2.11), but were only marginally more likely to undergo coronary artery bypass grafting (RR, 1.17; P<0.001; 95% CI, 1.07-1.28). CONCLUSIONS: In Victoria, management of patients with acute myocardial infarction is influenced by the public or private status of the patient, and by whether management occurs in private or public hospitals. Patients are more likely to undergo coronary angiography and coronary artery revascularisation procedures in private hospitals. PMID- 11061398 TI - Costs, charges and revenues of elective coronary angioplasty and stenting: the public versus the private system. AB - OBJECTIVE: To calculate the costs of elective coronary angioplasty and stenting (CAS) in the public and private healthcare systems and to compare these costs with the charges levied and the revenues obtained. DESIGN: A prospective health economics study. SETTING: A tertiary care public hospital and a co-located tertiary care private hospital in the 12 months from February 1998. STUDY POPULATION: 186 consecutive patients (124 public, 62 private) undergoing elective CAS. MAIN OUTCOME MEASURES: Outcome of CAS; exact costs of CAS in the two hospitals; exact charges to private patients; estimated charges in a typical, not co-located, "industry standard private hospital"; estimated costs to the Federal Government of CAS in the public and private system. RESULTS: The immediate and six-month outcomes in the two groups were similar. The average cost of CAS in public patients was $5,516, compared with $5,844 in private patients. The length of stay, number of stents per case and use of nonstent consumables was similar for both groups. Average charges for CAS in patients in the co-located private hospital were $13,347, and estimated average charges for CAS in an industry standard private hospital were $14,978. Estimated current costs to the government for CAS in a public hospital, a co-located private hospital, and an industry standard private hospital were $5664, $5,394 and $6,201, respectively. CONCLUSIONS: Despite similar treatments and similar treatment costs, CAS in the private system, as a consequence of the charges levied, is more than twice as expensive as in the public system, with government costs similar for both systems. These data (together with data from other studies showing that CAS is performed more frequently in private patients) suggest that encouraging more people to take out private health insurance will, paradoxically, increase government costs for CAS as well as increasing overall health expenditure. PMID- 11061399 TI - Cancer among people living in rural and remote Indigenous communities in Queensland. AB - OBJECTIVE: To describe the pattern of cancer among people living in rural and remote Indigenous communities in Queensland and to consider what implications the results have for cancer control. DESIGN AND SETTING: Descriptive analysis of data on incidence and mortality from the population-based Queensland Cancer Registry for the years 1982-1996. MAIN OUTCOME MEASURES: Age-standardised incidence and mortality rates for different cancer sites. RESULTS: The pattern of cancer was different from that found in the Queensland population as a whole. Of all the cancer sites, cervical cancer showed the biggest difference: the age-standardised incidence was 4.7 times the State average (95% CI, 3.2-6.6) and the mortality rate was 13.4 times the State average (95% CI, 7.8-21.4). Rates of lung cancer and other smoking-related cancers, although not as high as those for cervical cancer, were also significantly higher than the Queensland average, while rates for prostate and colorectal cancer were significantly lower. CONCLUSION: The cancers that are over-represented among Indigenous people are amenable to preventive measures. The cancer burden among Indigenous people could be reduced by lowering the prevalence of smoking and improving participation in cervical cancer screening and follow-up of screening-detected abnormalities. PMID- 11061400 TI - Three fatal pneumococcal polysaccharide vaccine failures. AB - Three Indigenous Australian adults died of invasive pneumococcal disease, despite vaccination with the pneumococcal polysaccharide vaccine an average of 18 months previously. Pneumococcal isolates from the patients had serotypes that are included in the vaccine. Two of the adults were alcoholics and the other had heavy recent alcohol intake. The public health implications, and possible optimal vaccination schedules to minimise vaccine failures, are discussed. PMID- 11061401 TI - The "Sherman effect": decreased ambulatory care volumes in Atlanta during the 1996 Summer Olympic Games. AB - OBJECTIVE: To estimate the effect of the 1996 Atlanta Summer Olympic Games on visits to local ambulatory healthcare facilities. DESIGN: Comparison of median visit rates by time period, obtained from retrospective review of administrative data. SETTING: The emergency department of the designated athletes' hospital, the public hospital's adult emergency department and adult walk-in clinics, and the adult and paediatric outpatient facilities of a large health maintenance organisation. PATIENTS: All 132,826 visitors to the designated facilities during the study interval. MAIN OUTCOME MEASURE: Daily visit frequencies at each facility. Our informal observations had suggested that volumes were not as high as expected. RESULTS: In all but the athletes' designated hospital, there was a decrease in average volumes the week before the opening ceremonies, ranging from zero to 8.4% of baseline. Average daily volumes in these non-venue facilities varied from 3.2% above to 16.1% below baseline during the two weeks of the Games, but all experienced an increase in volumes the week after the closing ceremonies, ranging from 3.0% to 13.7% of baseline. CONCLUSION: Unlike the venue-related facility, community ambulatory care sites did not encounter a significant rise in volumes until after the closing ceremonies. Although confirmation from other events is needed, our data suggest that, in addition to increased preparedness for sudden volume surges, overtime staffing of local facilities during planned mass gatherings should occur not during, but immediately after, the event. PMID- 11061402 TI - Drug testing at the Sydney Olympics. AB - With pre-Olympic and out-of-competition testing, as well as a new, validated test for erythropoietin, athletes will be exposed to more comprehensive drug testing at the Sydney Olympics. PMID- 11061403 TI - Newer drugs used to enhance sporting performance. AB - Controversy surrounding drug use in sport makes this a difficult area for rigorous research. However, it is striking that what data there are on drugs currently used for performance enhancement rarely indicate any clear benefit. PMID- 11061404 TI - Monitoring acute diseases during the Sydney 2000 Olympic and Paralympic Games. AB - The Sydney 2000 Olympic Games (the XXVII Olympiad) will be the biggest peacetime event ever held in Australia. During the Games, all public health decisions will be centralised, with daily briefing sessions held to review emerging public health issues and facilitate responses. Infectious diseases will be monitored and reported through the Olympic Surveillance System, with particular attention to foodborne diseases and conditions spread via the respiratory route. This system relies heavily on the cooperation of key notifiers such as emergency departments, laboratories and general practitioners. The lessons learned during the Games, and the new and enhanced systems and linkages that have been developed to support it, will strengthen future disease surveillance in NSW. PMID- 11061405 TI - Anabolic-androgenic steroids: medical assessment of present, past and potential users. AB - OBJECTIVE: To document adverse effects of anabolic-androgenic steroid (AAS) use in community-based users attending a medical clinic. DESIGN AND SETTING: Prospective recruitment, questionnaire-based interview, physical examination and investigations, with follow-up, of people who attended, anonymously, an inner city hospital clinic established specifically to examine AAS use. PARTICIPANTS: 58 men, comprising 27 past AAS users, 14 present users and 17 potential users (who formed the control group). MAIN OUTCOME MEASURE: Clinical adverse effects and abnormal laboratory findings. RESULTS: Cyclical use of oral and intramuscular, human and veterinary AASs were reported. The most commonly reported source of AASs was friends (59%), gymnasiums (25%) and doctors (14%). The most common reported adverse effects were alterations in libido (61%), changes in mood (48%), reduced testis volume (46%) and acne (43%). Although mean systolic and diastolic blood pressure was not significantly different between groups, five present (29%), 10 past (37%) and one potential user (8%) were hypertensive. Gynaecomastia was found in 10 past users (37%; P<0.01 v. potential users), two present users (12%) and no potential users. Mean testis volume was significantly smaller in present users (18 mL; P<0.02) than in the other groups. Twenty past users (83%), eight present users (62%) and five potential users (71%) had abnormal liver function test results (P=0.5). After discussion of test results, only 11 participants (19%) reported they would not use AASs in the future. CONCLUSIONS: Adverse effects were reported by or detected in most of the AAS users who attended the clinic. Despite awareness of adverse consequences, most participants planned future use of AASs. PMID- 11061406 TI - The health needs of people with intellectual disability. PMID- 11061407 TI - Adenoma size is important. PMID- 11061408 TI - Evidence to support colonoscopic screening is insufficient. PMID- 11061409 TI - Are Australian surgeons convinced about colorectal cancer screening? PMID- 11061410 TI - FOBT is being superseded. PMID- 11061411 TI - Conference overemphasised FOBT. PMID- 11061412 TI - Determining the validity of advance directives. PMID- 11061413 TI - The rise of embryology in Italy: from the Renaissance to the early 20th century. PMID- 11061414 TI - The "Stazione Zoologica Anton Dohrn" and the history of embryology. PMID- 11061415 TI - Molecularizing embryology: Alberto Monroy and the origins of Developmental Biology in Italy. PMID- 11061416 TI - Valerio Monesi, a pioneer of modern Reproductive Biology. PMID- 11061417 TI - On the present situation of Developmental Biology in Italy. PMID- 11061418 TI - From a home-made laboratory to the Nobel Prize: an interview with Rita Levi Montalcini. PMID- 11061419 TI - From human to sea urchin development: an interview with Giovanni Giudice. PMID- 11061420 TI - Regulation of primordial germ cell development in the mouse. AB - Primordial germ cells (PGCs) are the founders of the gametes. They arise at the earliest stages of embryonic development and migrate to the gonadal ridges, where they differentiate into oogonia/oocytes in the ovary, and prospermatogonia in the testis. The present article is a review of the main studies undertaken by the author with the aim of clarifying the mechanisms underlying the development of primordial germ cells. Methods for the isolation and purification of migratory and post-migratory mouse PGCs devised in the author's laboratory are first briefly reviewed. Such methods, together with the primary culture of PGCs onto suitable cell feeder layers, have allowed the analysis of important aspects of the control of their development, concerning in particular survival, proliferation and migration of mouse PGCs. Compounds and growth factors affecting PGC numbers in culture have been identified. These include survival anti apoptotic factors (SCF, LIF) and positive regulators of proliferation (cAMP, PACAPs, RA). Evidence has been provided that the motility of migrating PGCs relies on integrated signals from extracellular matrix molecules and the surrounding somatic cells. Moreover, homotypic PGC-PGC interaction has been evidenced that might play a role in PGC migration and in regulating their development. Several molecules (i.e. integrins, specific types of oligosaccharides, E-cadherin, the tyrosine kinase receptor c-kit) have been found to be expressed on the surface of PGCs and to mediate adhesive interactions of PGCs with the extracellular matrix, somatic cells and neighbouring PGCs. PMID- 11061421 TI - Female sterile mutations and egg chamber development in Drosophila melanogaster. AB - Drosophila oogenesis provides an excellent opportunity to study fundamental aspects of developmental biology and to learn the importance of multiple signalling pathways in the regulation of cellular morphogenesis. Taking advantage of the genetic and molecular approaches extremely powerful in this organism, over the years an enormous collection of data has accumulated on the genes involved in important steps of egg chamber development, such as germline and somatic stem cell maintenance, division and differentiation; oocyte determination and positioning; establishment of follicle cell fate and axes formation. These different processes are mediated by a reciprocal cross-talk between germline and somatic follicle cells. Here, in a schematic and simplified form, we point out what we believe are the main recent results on the molecular and cellular mechanisms underlying ovarian development and outline our recent contribution to this field. PMID- 11061422 TI - Paracrine actions of oocytes in the mouse pre-ovulatory follicle. AB - In mammals, ovulation requires a tight control of extracellular matrix modifications, within both the follicle wall and the inner mass of granulosa cells surrounding the oocyte, namely the cumulus cells. During the pre-ovulatory period, mural granulosa cells promote selective degradation of perifollicular matrix, resulting in the formation of a follicle rupture site. Conversely, cumulus cells synthesize a large amount of a muco-elastic matrix that plays an essential role in the extrusion of the oocyte from the follicle and in the subsequent fertilization process. Formation of such matrix by cumulus cells in the pre-ovulatory follicle appears to be controlled by a paracrine influence by the oocyte. We have shown that mouse oocytes modulate the response of cumulus cells to an ovulatory gonadotropin stimulus by promoting the synthesis and preventing the degradation of cumulus matrix. Therefore, although gonadotropins are essential for triggering the complex events involved in ovulation, the oocyte appears to have an active role in this process. In the present review current data and hypotheses concerning molecular mechanisms involved in the organization and synthesis of cumulus matrix are discussed. PMID- 11061423 TI - The role of stem cell factor and of alternative c-kit gene products in the establishment, maintenance and function of germ cells. AB - The c-kit gene plays a fundamental role during the establishment, the maintenance and the function of germ cells. In the embryonal gonad the c-kit tyrosine kinase receptor and its ligand Stem Cell Factor (SCF) are required for the survival and proliferation of primordial germ cells. In the postnatal animal, c-kit/SCF are required for the production of the mature gametes in response to gonadotropic hormones, i.e. for the survival and/or proliferation of the only proliferating germ cells of the testis, the spermatogonia, and for the growth and maturation of the oocytes. Finally, a truncated c-kit product, tr-kit, specifically expressed in post-meiotic stages of spermatogenesis and present in mature spermatozoa, causes parthenogenetic activation when microinjected into mouse eggs, suggesting that it might play a role in the final function of the gametes, fertilization. PMID- 11061424 TI - Sperm-egg interaction at fertilization: glycans as recognition signals. AB - This article first examines the events occurring in male and female genital tracts, which prepare human sperm to encounter the egg. Central is a glycoprotein, gp20, homologous to the leukocyte antigen CD52. This protein is secreted in the epididymal cells, inserted in the sperm plasma membrane and exposed in the equatorial region of the head at the end of the capacitation process. The mechanisms and molecules of the first interaction event between gametes in the mollusk bivalve Unio elongatulus and the current state of our knowledge of the same interaction in other species is then considered. The egg of Unio is very peculiar because it is highly polarized. Similar to other well-known egg models, the ligand for recognition is located on the egg coat which is a sort of fibrous network made up of very few glycoproteins, while the receptor is on the sperm surface. The difference is that in this egg, the ligand molecules are not uniformly distributed but are restricted to an area of the egg coat at the vegetal pole, the crater area. The role of carbohydrates in ligand function and of a specific type of oligosaccharide chain in particular, is discussed in the wider context of glycans acting as recognition signals. PMID- 11061425 TI - Multiple functions of Dlx genes. AB - Dlx genes comprise a highly conserved family of homeobox genes homologous to the distal-less (Dll) gene of Drosophila. They are thought to act as transcription factors. All Dlx genes are expressed in spatially and temporally restricted patterns in craniofacial primordia, basal telencephalon and diencephalon, and in distal regions of extending appendages, including the limb and the genital bud. Most of them are expressed during morphogenesis of sensory organs and during migration of neural crest cells and interneurons. In addition, Dlx5 and Dlx6 are expressed in differentiating osteoblasts. Gene targeting of Dlx1, Dlx2, Dlx3 and Dlx5 in the mouse germ-line has revealed functions in craniofacial patterning, sensory organ morphogenesis, osteogenesis and placental formation. However, no effect on limb development has yet been revealed from gene inactivation studies. A role for these genes in limb development is however suggested by the linkage of the Split Foot/Hand Malformation human syndrome to a region containing DLX5 and DLX6. As for most transcription factors, these genes seem to have multiple functions at different stages of development or in different tissues and cell types. PMID- 11061426 TI - Homeobox genes in the genetic control of eye development. AB - Vertebrate eye formation is a complex process which involves early specification of the prospective eye territory, induction events, patterning along the polarity axes and regional specification, to bring about the proper morphogenetic movements, cell proliferation, cell differentiation and neural connections allowing visual function. The molecular machinery underlying such complex developmental events is presently under an intense research scrutiny and many associated genetic factors have been isolated and characterized. These studies produced striking knowledge in the field, especially with respect to uncovering the role of key genes and their possible evolutionary conservation. Presently, a major task is to define the complex interactions connecting the multiplicity of molecular players that regulate eye development. We recently identified two homeobox genes, Xrx1 and Xvax2, and studied their function by using the Xenopus embryo as a developmental model system. Xrx1 and Xvax2 control key aspects of eye development. In particular, Xrx1 appears to play a role in the early specification of anterior neural regions fated to give rise to retina and forebrain structures, and in promoting cell proliferation within these territories. On the other hand, Xvax2 is involved in regulating the eye proximo distal and/or dorsoventral polarity, and the morphogenetic movements taking place during formation of the optic stalk and cup. Here we review the experimental results addressing the roles of Xrx1 and Xvax2 and their vertebrate orthologues, and discuss their relationship with other molecules also playing a related function in eye development. PMID- 11061427 TI - Homeobox genes and sea urchin development. AB - We describe the expression of three Paracentrotus lividus homeobox-containing genes of the dispersed class during sea urchin embryogenesis and discuss their possible roles in the mechanisms of cell specification and embryo morphogenesis. PlHbox12 represents the first regulator identified in sea urchin that belongs to the zygotic class of transcription factors. Its early and transient expression and the localization of transcripts suggests that PlHbox12 is involved in cell fate specification of the oral or aboral ectodermal territories at the early cleavage stages. PlHbox9 is expressed just after the completion of gastrulation in a narrow stripe of cells at the ectoderm-endoderm boundary. It probably organizes a novel spatial boundary which definitely separates the archenteron and the aboral ectoderm. Finally, the spatial and temporal expression of the PlOtp gene strongly indicate that this regulator is conditionally activated in few cells of the oral ectoderm and is involved in patterning of this territory at late stages. Furthermore, our data indicate that PlOtp acts upstream of signaling systems that lead to the activation of the primary mesenchyme cell gene expression program and skeletal morphogenesis. PMID- 11061428 TI - Met signaling mutants as tools for developmental studies. AB - The Met receptor is widely expressed in embryonic and adult epithelial tissues; its ligand (hepatocyte growth factor/scatter factor, HGF/SF) is expressed in the mesenchymal component of various organs. The generation of hgf and met null mice has revealed an essential role for this ligand-receptor pair in the development of the placenta, liver, and limb muscles. However the early lethality of the null mutants has precluded analysis of Met function in late development. To extend the possible observation period, we generated mutant metalleles of different severity. This was done by impairing the ability of the receptor to transduce the HGF/SF signal, via mutation of consensus sequences in the multifunctional docking site present in the C-terminal tail of the receptor. Mice expressing a Met mutant still active as a kinase, but unable to recruit its effectors, died in mid gestation with the same phenotype as the metknockout, proving the importance of phosphotyrosine-SH2 interactions in vivo. Mice expressing a Met receptor with partial loss of signaling function survived until birth and revealed novel aspects of HGF/SF-Met function during muscle development. PMID- 11061429 TI - "Tissue" transglutaminase in animal development. AB - The "tissue" transglutaminase is a multifunctional enzyme that in its cross linking configuration catalyzes Ca2+ -dependent reactions resulting in post translational modification of proteins by establishing epsilon(gamma-glutamyl) lysine cross-links and/or covalent incorporation of biogenic amines (di- and poly amines and histamine) into proteins. Several laboratories have shown that in Vertebrates, "tissue" transglutaminase (tTG) gene expression specifically characterizes cells undergoing apoptosis or programmed cell death (PCD). The Ca2+ -dependent activation of this enzyme leads to the formation of detergent insoluble cross-linked protein polymers in cells undergoing PCD. This insoluble protein scaffold could stabilize the integrity of the dying cells before their clearance by phagocytosis, preventing the non-specific release of harmful intracellular components (e.g. lysosomal enzymes, nucleic acids, etc.) and consequently inflammatory responses and scar formation in bystander tissues. In this review we attempt to present an overview of the current knowledge on tTG expression and regulation in animal reproduction and development. The data available so far further strengthen the relationship existing between tTG expression and the induction of PCD. PMID- 11061430 TI - Otx and Emx homeobox genes in brain development. AB - Over the last few years great progress has been made in the understanding of the formation and regionalisation of the mouse brain. In this review we will focus our attention on two families of homeobox-containing genes essentially coding for four transcription factors involved in brain and forebrain development: the two Emx and the two Otx genes. Here we describe the expression pattern of these genes in the developing mouse, as well as the characterisation of the corresponding knockout mice with special emphasis on Emx2. Whereas Otx genes are clearly involved in the formation and regionalisation of the whole rostral brain, comprised of forebrain and midbrain, our data suggest a role for Emx2 in the specification of the cytoarchitecture of the cerebral cortex, achieved through the control of proliferation of neuronal precursors and of migration of newly formed neurons to their final destination. PMID- 11061431 TI - The role of Otx and Otp genes in brain development. AB - Over the last ten years, many genes involved in the induction, specification and regionalization of the brain have been identified and characterized at the functional level through a series of animal models. Among these genes, both Otx1 and Otx2, two murine homologues of the Drosophila orthodenticle (otd) gene which encode transcription factors, play a pivotal role in the morphogenesis of the rostral brain. Classical knock-out studies have revealed that Otx2 is fundamental for the early specification and subsequent maintenance of the anterior neural plate, whereas Otx1 is mainly necessary for both normal corticogenesis and sense organ development. A minimal threshold of both gene products is required for correct patterning of the fore-midbrain and positioning of the isthmic organizer. A third gene, Orthopedia (Otp) is a key element of the genetic pathway controlling development of the neuroendocrine hypothalamus. This review deals with a comprehensive analysis of the Otx1, Otx2 and Otp functions, and with the possible evolutionary implications suggested by the models in which the Otx genes are reciprocally replaced or substituted by the Drosophila homologue, otd. PMID- 11061432 TI - Genetic and epigenetic control of midbrain dopaminergic neuron development. AB - The relatively few dopaminergic (DA) neurons in the mammalian brain regulate many important neural functions, including motor integration, neuroendocrine hormone release, cognition, emotive behaviors and reward. A number of laboratories, including ours, have contributed to unravel the mechanisms of DA phenotype induction and maturation and elucidated the role of epigenetic factors involved in specification, development and maintenance of midbrain dopaminergic functions. DA progenitors are first "committed" to give rise to DA neurons by the action of two secreted factors, Sonic hedgehog and fibroblast growth factor 8 (FGF8). Subsequently, the function of selectively activated transcription factors, Nurr1 and Ptx3, is required for the DA final determination. Further development of DA neurotransmission requires specific interactions with the developing target striatal cells, which modulate key DA functions, namely synthesis and uptake of the neurotransmitter. Committed and determined DA neurons express the key genes involved in DA neurotransmission at different times in development. In rodents, synthesis and intracellular accumulation of DA is achieved shortly after expression of Nurr1, while the onset of high affinity uptake, responsible for ending the neurotransmission, takes place after a few days. Cell contacts between the presynaptic DA neurons and target striatal neurons are apparently necessary for the fine modulation of DA function, in vivo and in vitro. Strikingly, the in situ maturation and phenotypic specialization of DA neurons grafted into the adult striatum/caudate-putamen parallels the normal development of committed fetal dopamine neurons during neurogenesis. The correct matching between the right presynaptic and postsynaptic neurons is required also for grafted DA cells. PMID- 11061433 TI - Acetylcholine synthesis and neuron differentiation. AB - Development of the nervous system is dependent on the co-operation between cell determination events and the action of epigenetic factors; in addition to well known factors, e.g. growth factors, neurotransmitters have been assigned a role as "morphogens" and modulators of neuronal differentiation in an early developmental phase. The possible role of acetylcholine as a modulator of neuronal differentiation has been considered in two experimental systems. A neuroblastoma cell line, which does not synthesise any neurotransmitter, has been transfected with a choline acetyltransferase construct; activation of acetylcholine synthesis, thus achieved, is followed by a higher expression of neuronal specific traits. The presence in these cells of muscarinic receptors is consistent with the existence of an autocrine loop, which may be responsible for the more advanced differentiation state observed in the transfected cells. Expression of cholinergic markers appears as a common feature of DRG sensory neurons, independently of the neurotransmitter used. Choline acetyltransferase can be detected in DRG at early developmental stages. The distribution of muscarinic receptors in DRG has suggested that activation of acetylcholine synthesis may be related in an early developmental phase to the interaction between neurons and nonneuronal cells and to modulation of cell differentiation. Both systems suggest that acetylcholine may have a role as a modulator of neuronal differentiation. PMID- 11061434 TI - Determination, diversification and multipotency of mammalian myogenic cells. AB - In amniotes, myogenic commitment appears to be dependent upon signaling from neural tube and dorsal ectoderm, that can be replaced by members of the Wnt family and by Sonic hedgehog. Once committed, myoblasts undergo different fates, in that they can differentiate immediately to form the myotome, or later to give rise to primary and secondary muscle fibers. With fiber maturation, satellite cells are first detected; these cells contribute to fiber growth and regeneration during post-natal life. We will describe recent data, mainly from our laboratory, that suggest a different origin for some of the cells that are incorporated into the muscle fibers during late development. We propose the possibility that these myogenic cells are derived from the vasculature, are multi-potent and become committed to myogenesis by local signaling, when ingressing a differentiating muscle tissue. The implications for fetal and perinatal development of the whole mesoderm will also be discussed. PMID- 11061435 TI - Developmental control of chondrogenesis and osteogenesis. AB - During vertebrate embryogenesis, bones of the vertebral column, pelvis, and upper and lower limbs, are formed on an initial cartilaginous model. This process, called endochondral ossification, is characterized by a precise series of events such as aggregation and differentiation of mesenchymal cells, and proliferation, hypertrophy and death of chondrocytes. Bone formation initiates in the collar surrounding the hypertrophic cartilage core that is eventually invaded by blood vessels and replaced by bone tissue and bone marrow. Over the last years we have extensively investigated cellular and molecular events leading to cartilage and bone formation. This has been partially accomplished by using a cell culture model developed in our laboratory. In several cases observations have been confirmed or directly made in the developing embryonic bone of normal and genetically modified chick and mouse embryos. In this article we will review our work in this field. PMID- 11061436 TI - Role of the extracellular matrix and growth factors in skull morphogenesis and in the pathogenesis of craniosynostosis. AB - The complex and largely obscure regulatory processes that underlie ossification and fusion of the sutures during skull morphogenesis are dependent on the conditions of the extracellular microenvironment. The concept that growth factors are involved in the pathophysiology of craniosynostosis due to premature fusion of skull bone sutures, is supported by recent genetic data. Crouzon and Apert syndromes, for example, are characterized by point mutations in the extracellular or transmembrane domains of fibroblast growth factor-2 receptor. In primary cultures of periosteal fibroblasts and osteoblasts obtained from Apert and Crouzon patients, we observed that Crouzon and Apert cells behaved differently with respect to normal cells as regards the expression of cytokines and extracellular matrix (ECM) macromolecule accumulation. Further modulation of ECM components observed after the addition of cytokines provides support for an autocrine involvement of these cytokines in Crouzon and Apert phenotype. Changes in ECM composition could explain the altered osteogenic process and account for pathological variations in cranial development. We suggest that a correlation exists between in vitro phenotype, clinical features and genotype in the two craniosynostotic syndromes. New research into signal transduction pathways should establish further connections between the mutated genotype and the molecular biology of the cellular phenotype. PMID- 11061437 TI - Integrin function and regulation in development. AB - Integrins are a large family of membrane receptors, consisting of alpha and beta subunits, that play a pivotal role in the interaction of cells with the extracellular matrix. Such interaction regulates the organization of cells in organs and tissues during development as well as cell differentiation and proliferation. We have shown that unfertilized oocytes express integrins that might be important during fertilization. We also analyzed nervous system and muscle tissue development showing that integrin expression is precisely regulated during organization of these tissues. The results indicate that two distinct integrin alpha subunits mediate the outgrowth of processes in nerve and glial cells. Alpha1 integrin, a laminin receptor, is up-regulated by nerve growth factor and other differentiation stimuli and is involved in neurite extension by nerve cells. In contrast, process extension by glial cells is likely to involve the alphaV integrin. Moreover, the latter integrin subunit is also transiently expressed in muscle of the embryo body where it localizes predominantly at developing myotendinous junctions. After birth this integrin disappears and is substituted by the alpha7 subunit. At the same time, important changes also occur in the expression of the associated beta subunit. In fact, the beta1A isoform which is expressed in fetal muscles, is substituted by beta1D. These isoforms are generated by alternative splicing and differ in only a few amino acid residues at the COOH terminus of the protein. This region of the molecule is exposed at the cytoplasmic face of the plasma membrane and is connected to the actin filaments. Our results show that beta1D, which is expressed only in striated muscle tissues, binds to both cytoskeletal and extracellular matrix proteins with an affinity higher than beta1A. Thus, beta1D provides a stronger link between the cytoskeleton and extracellular matrix necessary to support mechanical tension during muscle contraction. These results indicate that cells can regulate their interactions with the extracellular matrix by changing their expression of alpha integrin subunits and thus ligand specificity, or by more subtle changes involving alternative usage of different cytoplasmic domains. The important role of both alpha and beta integrin subunit cytoplasmic domains during development is further illustrated by the analysis of targeted mutations which we have generated by homologous recombination in mice. PMID- 11061438 TI - Cell-cell signaling and adhesion in phagocytosis and early development of Dictyostelium. AB - Cell-cell signaling and adhesion regulate transition from the unicellular to the multicellular stage of development in the cellular slime mold Dictyostelium. Essential gene networks involved in these processes have been identified and their interplay dissected. Heterotrimeric G protein-linked signal transduction plays a key role in regulating expression of genes mediating chemotaxis or cell adhesion, as well as coordinating actin-based cell motility during phagocytosis and chemotaxis. Two classes of cell adhesion molecules, one cadherin-like and the second belonging to the IgG superfamily, contribute to the strength of adhesion in Dictyostelium aggregates. The developmental role of genes involved in motility and adhesion, and their degree of redundancy, have been re-assessed by using novel developmental assay conditions which are closer to development in nature. PMID- 11061439 TI - The molecular organization of endothelial junctions and their functional role in vascular morphogenesis and permeability. AB - We review here our work on the molecular and functional organization of endothelial cell-to-cell junctions. The first part of the review is dedicated to VE-cadherin, characterized by our group few years ago. This protein is a member of the large family of transmembrane adhesion proteins called cadherins. It is endothelial cell specific and plays a major role in the organization of adherens junctions. Inactivation of VE-cadherin gene or in vivo truncation of its cytoplasmic tail leads to a lethal phenotype due to the lack of correct organization of the vasculature in the embryo. We found that the defect was due to apoptosis of endothelial cells, which became unresponsive to the survival signal induced by vascular endothelial cell growth factor. Our data indicate that VE-cadherin may act as a scaffolding protein able to associate vascular endothelial cell growth factor receptor and to promote its signaling. In the second part of the review we consider another protein more recently discovered by us and called junctional adhesion molecule (JAM). This protein is a small immunoglobulin which is located at tight junctions in the endothelium and in epithelial cells. Evidence is discussed indicating that JAM takes part in the organization of tight junctions and modulates leukocyte extravasation through endothelial intercellular junctions in vitro and in vivo. The general role of tight junctions in endothelial cells is also discussed. PMID- 11061440 TI - Acetabular reconstruction with impacted morcellized cancellous bone autograft and cemented primary total hip arthroplasty: a 10- to 17-year follow-up study. AB - During the period 1979 through 1986, 69 acetabular reconstructions in 63 patients were performed with the use of autologous morcellized bone-grafts because of acetabular bone stock loss. Nine cases (10 hips) were lost to follow-up. Eleven patients (12 hips) died <10 years after surgery; none had a revision. The results for the remaining 43 patients (47 hips) were reviewed at an average interval of 12.3 years (range, 10-17 years). No preoperative Harris hip score was available. The average Harris hip score at follow-up was 88 (range, 60-100). Radiographically, all grafts united. One hip developed a deep infection. Three other hips (6%) were revised because of aseptic loosening of the acetabular component. An additional 3 acetabular components were considered radiographic failures. Excluding the infected case, the overall survival rate of these acetabular reconstructions with a revision as endpoint was 94% at an average follow-up of 12.3 years. Reconstruction of acetabular bone stock loss with autologous morcellized bone-grafts is an attractive technique with a good potential for long-term success. PMID- 11061441 TI - Low-friction arthroplasty in patients younger than 40 years old: 20- to 25-year results. AB - A total of 67 low-friction arthroplasties were performed from 1971 through 1978 in patients <40 years old and followed an average of 21.7 years. There have been 33 cup loosenings and 17 stem loosenings. The average rate of wear of the cup was 0.12 mm/y for the entire series and 0.16 mm/y for the revised cups. Although the preoperative diagnosis leading to low-friction arthroplasty commonly reflected some deficiency in bone structure, particularly in the acetabulum, there were no primary osteoarthrosis cases. The femoral stem proved to be durable, but acetabular cups gave poorer results in younger patients than in older patients. The 2 major factors limiting the longevity of the cemented cup were acetabular bone quality and wear. PMID- 11061442 TI - Cementless primary total hip arthroplasty with a tapered, proximally porous coated titanium prosthesis: a 4- to 8-year retrospective review. AB - A consecutive series of 100 primary total hip arthroplasties were performed at a single institution on 87 patients using a cementless collared titanium press-fit stem. Of patients, 87% received a hemispheric porous-coated cup, and 13% received a nonmodular titanium fibermesh press-fit cup. Ten hips were excluded from the longer-term evaluation: 6 were lost to follow-up, and 4 patients were deceased. Ninety hips, with an average follow-up of 81 +/- 12 months, were retrospectively reviewed. The average postoperative hip score was 94, compared with an average preoperative hip score of 42. No postoperative infections were observed, but there were 2 cases of postoperative dislocation (2%) and 1 case of thigh pain (1%) at last follow-up. There were 2 revisions, both for cup failures. There were no femoral component loosenings or revisions. There was no evidence of stem subsidence or instability. These midterm results are encouraging with this stem design. PMID- 11061443 TI - Clinical experience with a modular noncemented femoral component in revision total hip arthroplasty: 4- to 7-year results. AB - A series of 163 revision total hip arthroplasties performed using a modular proximally porous-coated device was reviewed. Twenty patients died before achieving minimum follow-up, and 13 hips with Paprosky type IV femora were excluded. One patient was lost to follow-up. Minimum 4-year clinical data on the remaining 129 hips showed an improvement in modified Harris hip scores, from an average of 47.7 to 87.5. Spot welds at the sleeve-bone interface were found in 83 of the 102 hips with minimum radiographic follow-up. The aseptic failure rate was 2.9%; these hips showed progressive subsidence, with 1 resulting in the only repeat revision in the series. Three hips had osteolytic lesions of <5 mm; none threatened implant stability. There were no complications related to modularity. PMID- 11061444 TI - Reconstruction of major column defects and pelvic discontinuity in revision total hip arthroplasty. AB - Acetabular reconstruction with severe bone loss after failed total hip arthroplasty is a difficult problem. Defects were defined as major segmental and cavitary loss (type III anterior or posterior) or pelvic discontinuity (type IV). Seventeen cases were treated, of which 7 were type III and 10 were type IV. Bulk allograft was used in 16 of 17 cases, of which 7 were whole acetabular grafts, 2 were posterior segmental acetabular grafts, and 7 were femoral heads. Fourteen of 17 patients were female. The extensile triradiate approach was used in 12 cases. Long pelvic bone plates were applied to the posterior column and anterior brim of the pelvis in most cases. Allografts united to host-bone in 15 cases. Average follow-up was 83 months. The overall revision rate was 47%, of which 3 of 7 press fit and 2 of 10 cemented cups had failed. The dislocation rate for the extensile approach was 50%; 2 patients had excisional arthroplasty for infection, and 2 patients had exploration of the sciatic nerve for release from migrating pelvic plate screws. Because of the overall poor results, this approach cannot be recommended for general use. PMID- 11061445 TI - The use of a modular rotating hinge component in salvage revision total knee arthroplasty. AB - Revision total knee arthroplasty (TKA) using a second-generation modular rotating hinge design was performed on 16 knees in 15 patients over a 5-year period. Follow-up of 2 to 6 years (mean, 51 months) was obtained in 14 knees in 13 patients. Indications for revision were aseptic loosening of a hinged prosthesis (8 knees), loosening and bone loss associated with chronic extensor mechanism disruption (2 knees), component instability with chronic medial collateral ligament disruption (3 knees), and comminuted distal femur fracture (1 knee). Clinical and radiographic results were reviewed and compared with 87 patients who underwent revision TKA using a standard condylar revision design during the same period. Early results showed comparable postoperative knee scores and range of motion between the 2 groups despite the use of the rotating hinge component in more complex revision cases. No patient has exhibited radiographic evidence of definite component loosening. Alignment of 5 degrees to 10 degrees of valgus in the frontal plane and within 2 degrees of neutral in the sagittal plane was achieved consistently. Short-term clinical and radiographic results are encouraging and suggest that a second-generation modular rotating hinge component can be used successfully in selected salvage revision cases. PMID- 11061446 TI - Intraoperative electromyography of the superior gluteal nerve during lateral approach to the hip for arthroplasty: a prospective study of 12 patients. AB - The purpose of this study was to evaluate the incidence of intraoperative superior gluteal nerve irritation and to identify specific surgical maneuvers that may harm the nerve. Continuous intraoperative electromyography (EMG) monitoring of the superior gluteal nerve-innervated muscles (gluteus medius and tensor fascia lata muscles) was performed in 12 patients undergoing total hip arthroplasty. A modified lateral approach was used, including a partial anterior osteotomy of the greater trochanter with splitting of the gluteus medius and vastus lateralis muscles. All patients had a clinical follow-up examination 1 year postoperatively to evaluate abductor muscle function. Irritation of the nerve occurred first during splitting of the gluteus medius muscle, then with increased gluteus medius retraction for exposure of the acetabulum, and finally during positioning of the leg for preparation of the femur. The detected EMG alterations were important because they were found in a single patient with persistent abductor muscle weakness. PMID- 11061447 TI - Arthrodesis of the knee: experience with intramedullary nailing. AB - Knee arthrodesis using an intramedullary nail has gained acceptance as treatment in difficult cases such as infection after total knee arthroplasty (TKA), neuropathic joint, and obesity. A retrospective review of 22 cases treated at our institution using an intramedullary nail for knee arthrodesis was performed. Deep infection after primary (11) or revision (6) TKA was the most common indication for this procedure. A long intramedullary nail was used in 3 cases, a long nail with a proximal interlocking screw was used in 6 cases, and a customized nail with a valgus bend and a proximal interlocking screw was used in 11 cases. A modular knee fusion nail was used in 1 case. Successful fusion occurred in all cases, although 4 patients required additional surgery. Average operative blood loss was 748 mL, and average time to union was 7 months. Shortening of the extremity averaged 3.2 cm. Tibiofemoral alignment was improved by using a customized valgus nail (average, 3.1 valgus; range, 1-5) when compared with a straight nail (average, 0.2 valgus; range, 3 varus to 3 valgus). No patient developed infection in the hip or ankle region as a result of the long intramedullary nail. Intramedullary nailing is an excellent technique for knee arthrodesis in difficult cases. A customized proximal interlocking nail with 5 degrees to 7 degrees of valgus and 5 degrees of anterior angulation improves tibiofemoral alignment and is straightforward to insert or extract should it be necessary. Stability and pain relief are rapid, and the fusion rate is maximized. PMID- 11061448 TI - The impact of health status on waiting time for major joint arthroplasty. AB - This study was conducted to determine the impact of health status on waiting time for major joint arthroplasty in a universal publicly funded health system. Data were collected prospectively from a cohort of 553 patients waiting for total hip or total knee arthroplasty. The WOMAC and SF-36 health status instruments were administered at the time the patient was placed on the waiting list. The outcome measure was length of waiting time. Multivariate analyses found increased body mass index and decreased social function as the only determinants of waiting time. This model explained only 4% of the variance in waiting time. The association between health status and waiting time for arthroplasties appears to be small. These findings suggest that major joint arthroplasty is not prioritized on the basis of burden of illness. PMID- 11061449 TI - Are the findings in the Swedish National Total Hip Arthroplasty Register valid? A comparison between the Swedish National Total Hip Arthroplasty Register, the National Discharge Register, and the National Death Register. AB - The Swedish National Total Hip Arthroplasty (THA) Register was initiated in 1979, and it is one of the oldest quality registers in the world. The register covers all hospitals in Sweden, and today it contains > 205,000 hip arthroplasties. The failure endpoint definition in the register is revision. There is no information about quality of life and mortality. The aim of this study was to validate the results presented by the Swedish THA register by comparison with the Discharge register (the Swedish National Board of Health and Welfare) and to study mortality after hip arthroplasties. All hip arthroplasties from the Discharge register, performed in 1986 and thereafter, were compared with the Swedish THA register. Epidemiologic parameters, including mortality, were documented from the Swedish Death register. The mortality for primary THAs for men was 1% higher and for women 6% higher when compared with an age-matched and sex-matched cohort. For revision, the numbers were 7% and 9% higher. The risk for death compared with an age-matched and sex-matched population was lower for patients with osteoarthrosis treated with hip arthroplasty. The results with revision as failure endpoint showed that the Swedish THA register is reliable. The register includes >95% of the primary and revision THAs performed in Sweden between 1986 and 1995. PMID- 11061450 TI - Radiographic anatomic structure of the arthritic acetabulum and its influence on total hip arthroplasty. AB - Acetabular bone structure is not the same in all patients and can be defined by the radiolucent triangle superior to the acetabulum. Of 132 hips, 81 had an isosceles triangular shape, which was named type A acetabulum. Forty-six hips had an extension of the triangle into the teardrop, which created a thickened medial wall and was named type B. Five hips had a right-angle triangle, which was found only with congenital disease of the hip and was named type C. The density of the superior acetabular bone in the triangle could be normally radiolucent (stage I), have vertical and transverse trabeculae throughout the triangle (stage II), or have the triangle filled with bone and cysts (stage III). The relationship between progressive radiolucent lines and acetabular type showed that type A3 (thin medial wall with dense triangle bone) had the highest incidence of progressive radiolucent lines (P < .05). PMID- 11061451 TI - Variation in the wear rate during the life of a total hip arthroplasty: a simulator and retrieval study. AB - The limitation of wear is fundamental to the optimization of total hip arthroplasty longevity. The maintenance of the supersmooth femoral head surface is considered to be paramount in maximizing prosthesis life expectancy. Ex vivo studies have failed to substantiate a relationship between roughness and the clinical wear factor, however. A hip simulator wear study was undertaken to investigate this contradiction. Three explanted femoral heads were articulated for 5 million cycles against new acetabular liners. The simulator wear rate was 5 times the ex vivo value. This difference can be explained only if the explant head roughness was not that which existed for most of the joint's life. The relationship between surface roughness and wear deduced for simulator testing is substantially different from that of unidirectional wear screening methods. The multiphasic nature of wear in cementless joints has been illustrated: a wear-in period, followed by a steady-state phase, until a head-roughening event causes a rapid wear period. PMID- 11061452 TI - Movement loci of selected points on the femoral head for individual total hip arthroplasty patients using three-dimensional computer simulation. AB - Wear of ultra-high-molecular-weight polyethylene (UHMWPE) and the subsequent adverse tissue reaction to the wear particles has been cited as the predominant problem affecting total joint arthroplasties. Wear of the UHMWPE acetabular cup in total hip arthroplasties (THAs) is influenced by the sliding distance and direction of individual points on the femoral head, which has as yet been ascertained only for normal subjects. This study seeks to determine the trajectory of specified points on the femoral head for individual THA patients and the distances traversed by these points. A computer program was designed to use gait data from THA patients to simulate the motion of these points. Gait analysis was performed on 19 THA patients at a period of 5 years after operation. The orientation of the acetabular cup was ascertained from radiographs to determine the position of the points on the head most involved in the wear process. The loci of the points differed widely in size, shape, and direction between subjects. The largest average distance traversed was 140% greater than the smallest average distance traversed. Shorter longitudinal paths would tend to cause less wear than larger, wider paths, which cross a greater number of adjacent paths. PMID- 11061453 TI - Effect of posterior cut angle on tibial component loading. AB - Although clinical studies have shown that posterior cut angle affects tibial component stability, biomechanical studies are lacking. Fifteen Sawbones tibiae were divided into 5 groups and prepared with 0 degrees , 3 degrees, 6 degrees, 9 degrees, and -5 degrees tibial surface cuts, and a tibial component was implanted with cement. Using a standard and then a highly congruent polyethylene insert, the knee was loaded at 0 degrees and 30 degrees of flexion. There were statistically significant increases in anterior micromotion of the standard polyethylene component for each increase in posterior slope cut angle, which increased for the highly congruent polyethylene component. The anterior slope cut (-5 degrees) led to significant posterior micromotion of the tibial polyethylene component. Increased posterior slope cut angle significantly decreased tibial anterior compressive strains and significantly increased tibial posterior compressive strains. The highly congruent insert significantly increased this posterior strain. The results indicate that cutting the articular surface of the tibia at a 0 degrees or 3 degrees posterior slope provides the greatest tibial component stability. PMID- 11061454 TI - Lavage technique in total hip arthroplasty: jet lavage produces better cement penetration than syringe lavage in the proximal femur. AB - Sixteen paired human cadaver femora were prepared using conventional broaches. Cancellous bone was irrigated with 1 L pulsed lavage in one femur and 1 L syringe lavage in the contralateral femur. The specimens were embedded in specially designed pots, and vacuum-mixed bone-cements were applied in a retrograde manner. After application of a standard pressure to the pots, the femora were removed and radiographed, and horizontal sections were obtained and analyzed to assess cement penetration into cancellous bone and the ratio of the area of supported to unsupported cancellous bone (Rcb). Our results show that in equal quality bone, the use of jet lavage yields significantly (P < .0001) improved cement penetration and Rcb compared with syringe lavage specimens. Jet lavage should be considered routine to achieve interdigitation with cancellous bone in cemented total hip arthroplasty. PMID- 11061455 TI - Long-term survival of the T-28 versus the TR-28 cemented total hip arthroplasties. AB - Between 1974 and 1980, 550 total hip arthroplasties (THAs) (479 patients) were performed using T-28 and TR-28 cemented prostheses (TR-28 is shot-blast chrome and T-28 is polished stainless steel). There were 379 cemented THAs in 321 patients in the T-28 group and 171 cemented THAs in 158 patients in the TR-28 group. Average follow-up of the patients still alive at the end of the study was 20.96 years in the T-28 group and 17.54 years in the TR-28 group. When considering failure as revision of the hip for aseptic acetabular loosening, there were 36 (9.5%) failures in the T-28 group and 12 (7%) failures in the TR-28 group. This difference was statistically significant (P = .0132). When considering failure as radiographic acetabular loosening with or without radiographic femoral loosening, there were 52 failed acetabula (13.7%) in the T 28 group and 18 failed acetabula (10.5%) in the TR-28 group. These differences were not statistically significant. When considering failure as revision for aseptic femoral loosening with or without acetabular component loosening, there were 42 failures (11.1%) in the T-28 group and 22 failures (12.8%) in the TR-28 group. This difference was not statistically significant. When considering failure as radiographic femoral loosening with or without acetabular component loosening, there were 42 failures (11.1%) in the T-28 group and 27 failures (15.8%) in the TR-28 group. This difference was statistically significant for log rank test (P = .0318) and Wilcoxon's test (P = .0083). Surface finish may be an important contributor to the survival of cemented femoral stems. PMID- 11061456 TI - Disassembly of a Howmedica modular resection system. AB - We report the postoperative disassociation of a Morse taper junction of a Howmedica Modular Resection system implanted after the resection of a malignant distal femoral bone tumor. The disassembly occurred without apparent trauma. This complication was managed by closed reduction under epidural anesthesia. This disassembly probably occurred because of decreased tension of the soft tissue around the prosthesis or an impaired locking mechanism. PMID- 11061457 TI - Serum titanium level for diagnosis of a failed, metal-backed patellar component. AB - A case is presented in which an elevated serum titanium level was used to make the diagnosis of a failed metal-backed patellar component. The preoperative serum titanium level was 536.8 ppb, which was 98 times higher than the patient's previous level (taken 1 year earlier, when he was asymptomatic) and 2 orders of magnitude higher than the expected level with a well-functioning implant of this type. Revision surgery confirmed that the polyethylene portion of the patellar component had worn through, leaving the titanium portion of the patellar implant to articulate with the femoral component. Wear-through was not evident on preoperative radiographs or clinical examination. As knowledge about the expected ranges for serum metal ion levels after total joint arthroplasty continues to increase, the diagnostic utility of serum metal ion testing in the evaluation of joint arthroplasty function will continue to improve. PMID- 11061458 TI - Stem displacement during reduction of a dislocated cemented total hip arthroplasty with a polished tapered stem. AB - Displacement of the polished stem of a total hip arthroplasty from its cemented mantle occurred as a complication of an attempted closed reduction after luxation. No cement fractures were seen during subsequent reoperation. The complication can be avoided if cement is placed over the shoulder of the prosthesis. PMID- 11061459 TI - Plugging the intramedullary canal of the femur in total knee arthroplasty: reduction in postoperative blood loss. AB - A prospective, randomized trial comparing postoperative drainage was carried out in 120 consecutive knees undergoing total knee arthroplasties divided into 2 groups. In one (55 knees), the entry point for the femoral intramedullary rod was left open. In the other (65 knees), the entry point was closed by an autologous bone plug. The mean drainage after 24 hours and the total drainage were lower when the femoral canal was plugged (800 vs 960 mL and 925 vs 1,165 mL). The bone plug always united, and no loose bodies were seen 6 months after operation. We conclude that the femoral intramedullary guide hole should be plugged with autologous bone because this technique results in a small but significant reduction in early blood loss without countervailing disadvantages. PMID- 11061460 TI - Wavelet-based speckle noise reduction in ultrasound B-scan images. AB - Speckle noise is known to be signal-dependent in ultrasound imaging. Hence, separating noise from signal becomes a difficult task. This paper describes a wavelet-based method for reducing speckle noise. We derive from the model of the displayed ultrasound image the optimal wavelet-domain filter in the least mean square sense. Simulations on synthetic data have been carried out in order to assess the performance of the proposed filter with regards to the classical wavelet shrinkage scheme, while phantom and tissue images have been used for testing it on real data. The results show that the filter effectively reduces the speckle noise while preserving resolvable details. PMID- 11061461 TI - Axial strain imaging using a local estimation of the scaling factor from RF ultrasound signals. AB - The main signal-processing techniques used in elastography compute strains as the displacement derivative. They perform well for very low deformations, but suffer rapidly from decorrelation noise. Aiming to increase the range of accurate strain measurements, we developed an adaptive method based on the estimation of strains as local scaling factors. Its adaptability makes this method appropriate for computing scaling factors resulting from larger strains or a wide spread of strain variations. First, segments corresponding to the same part of tissue are adaptively selected in the rest and stressed state echo signals. Then, local scaling factors are estimated by iteratively varying their values until reaching the zero of the phase of the complex cross-correlation function. Results from simulation and from experimental data are presented. They show how this adaptive method can track various local deformations and its accuracy for strain up to 7%. PMID- 11061462 TI - Interpolation of missing part of human follicle border on ultrasonic B-mode image by iterative revision. AB - In clinical infertility treatments, assessment of ovarian follicle growth by ultrasonography is important. In order to measure the geometrical characteristics of the human follicle, such as the area of the cross-sectional image and the volume inside the follicle, a method based on manual tracing of the follicle contour from the ultrasonic B mode image is widely used. However, the observable ultrasonic B mode images are sometimes imperfect and some parts of the follicle border are missing due to the existence of the acoustic shadow. In this paper, a method that interpolates the missing part of the follicle border from the known part is proposed. This method uses a priori information of the follicle, which is usually known in actual cases: (1) the follicle's surface is so smooth that its border is assumed to be a smooth closed curve; and (2) the position of the follicle's center is roughly predicted in advance in the ultrasonic B-mode image. In the proposed method, the missing part of the human follicle border is interpolated from the known part by applying an iterative revision so as to satisfy the smoothness condition of the follicle. This method is also applied to three-dimensional image reconstruction of the human follicle. PMID- 11061463 TI - Feasibility of biomedical applications of Hall effect imaging. AB - Hall effect imaging is a new technique for mapping the electrical properties of a sample. Its principle has been demonstrated in two- and three-dimensional phantom images. Based on the experimental data and theoretical understanding of this technique developed over the past few years, this paper addresses the most relevant question for biomedical applications: whether Hall effect imaging is ultimately applicable to complex biological systems such as the human body. The arguments are given at the basic physics level, so that the conclusion is independent of current technology status. These arguments are corroborated with imaging data of an aorta sample. The conclusion is that Hall effect imaging is not suited for quantifying the electrical constants in complex biological samples. This technique is able to produce high-resolution volume images of samples in vitro that reflect their electrical heterogeneity. However, quantitative measurements of electrical constants are not practical for complex samples. PMID- 11061464 TI - The trouble with sinuses. PMID- 11061465 TI - Dose-dependent effects of intranasal steroids: how relevant? PMID- 11061466 TI - Desensitization and test-dosing for the drug-allergic patient. PMID- 11061467 TI - Allergic vulvovaginitis. AB - LEARNING OBJECTIVES: The reader of this review will learn about the different clinical forms of allergic vulvovaginitis. This specific and important chapter has not been previously summarized and described in the medical literature. Vaginal mucosa is also able to show an allergic response similarly to the nose, eyes, lungs, and skin. Physicians should be familiarized with this kind of manifestation in order to make the proper diagnosis and evaluation of this entity. DATA SOURCES: MEDLINE searches were undertaken since 1966 for citations of any kind of allergic vulvovaginitis. Relevant reviews and articles identified in this process were surveyed for additional and earlier citations. Textbooks of medicine, gynecology, dermatology, and infectious diseases have also been consulted. Old medical textbooks and journals of allergy and internal medicine were recovered from the Division of History of the Medicine of the Faculdade de Medicina da Universidade Federal de Minas Gerais (Federal Medical College), Belo Horizonte, Brazil. CONCLUSIONS: A great variety of allergens are able to provoke allergic reactions in the female genital tract. The immunology of the vagina, the influence of hormones, menstrual cycle, and psychologic factors are also highlighted in this review. A possibility of vaginal hyperreactivity is proposed in this text. Adequate management provides important relief of symptoms in the majority of cases. PMID- 11061468 TI - Pseudoallergic toxic reaction. PMID- 11061469 TI - "Allergic sinusitis" revisited. AB - BACKGROUND: "Allergic sinusitis" is a frequently used term but there is question about its true existence. OBJECTIVE: To detect the presence of allergic inflammation of the sinuses in five highly symptomatic ragweed sensitive adults. METHOD: Three imaging techniques were utilized: SPECT bone imaging, SPECT Indium111 labeled WBC uptake, and FDG F-18 (PET scanning). RESULTS: We could find no evidence of sinus involvement in association with severe allergic rhinitis. CONCLUSIONS: Employing three different imaging modalities, we were unable to demonstrate inflammation of the sinuses in patients with allergic rhinitis. PMID- 11061470 TI - Dose-dependent effects of budesonide aqueous nasal spray on symptoms in a daily nasal allergen challenge model. AB - BACKGROUND: It has been difficult to demonstrate dose-dependent clinical effects of anti-allergic glucocorticosteroid drugs in allergic rhinitis. OBJECTIVE: To determine dose-dependent effects on rhinitis symptoms of clinical doses of the glucocorticosteroid budesonide in a standardized daily allergen challenge model. METHODS: Twenty-five patients with seasonal allergic rhinitis were examined outside the pollen season. The highest 256 microg once daily and lowest 64 microg once daily clinically recommended doses of budesonide aqueous nasal spray and placebo were given in a double-blind, placebo-controlled, randomized, and crossover design with 4 weeks' washout between treatments. After 1 week's treatment, the patients received individually titrated nasal allergen challenges once every morning for 8 days while treatment continued. Nasal symptoms were scored in diary cards. Nasal symptoms from the 6th to the 8th challenge days were used in the analysis. RESULTS: The provocation model produced clinically relevant, and around the clock well tolerable rhinitis symptoms, suggesting that after several days of repeated allergen challenges, a season-like, transient allergic disease condition had been established. Both 64 microg and 256 microg of budesonide aqueous nasal spray reduced nasal symptoms. Budesonide 64 microg reduced total nasal symptoms scores from 5.19 +/- 0.5 to 4.23 +/- 0.53 (P < .05), and budesonide 256 microg reduced total nasal symptoms scores to 3.41 +/- 0.51 (P < .001). A significant difference in nasal symptoms after challenge between budesonide aqueous nasal spray 64 microg and 256 microg (P = .03), indicated a dose-dependent effect. CONCLUSIONS: A dose-dependent, symptom-reducing effect of once-daily treatment with topical aqueous nasal sprays of budesonide for two weeks was demonstrated, suggesting that this model is relevant for assessments of dose-dependent effects of anti-inflammatory drugs. PMID- 11061471 TI - Passive dispersion of latex aeroallergen in a healthcare facility. AB - BACKGROUND: Prompted by worsening asthma in a dental assistant with latex allergy and occupational asthma while under personal latex precautions, we confirmed continuing latex aeroallergen exposure. OBJECTIVES: To determine the source of latex aeroallergen and ascertain the effects of site-wide substitution of nonpowdered low allergen latex glove in a health care site. METHODS: Using a volumetric sampler, baseline latex aeroallergen levels were measured in rooms where she worked and nearby rooms, as well as shared X-ray, laboratory, and waiting rooms. Allergen levels were measured in upholstery fabric samples, ventilation duct dust, and latex gloves. Alterations in aeroallergen levels following change of glove types were prospectively determined. RESULTS: Baseline latex aeroallergen levels ranged from 6 to 25 ng/m3 in the patient's work areas and in other rooms from 29 to 90 ng/m3 during work hours. Latex antigen was found in three brands of powdered latex gloves (chi = 1,156 microg/g) used in the nearby opertories and the hygiene room, and in upholstery fabric, carpet dust, but not ventilation duct dust. In the absence of any other control measures, airborne latex became undetectable (<5 ng/m3) with exclusive use of nonpowdered latex gloves. DISCUSSION: Latex aeroallergen is primarily generated by active glove use; carpeting and fabric upholstery can serve as important aeroallergen repositories. Site-wide substitution of nonpowdered latex gloves eliminates detectable latex aeroallergen. PMID- 11061472 TI - Atopy in Danish children and adolescents: results from a longitudinal population study. AB - BACKGROUND AND OBJECTIVE: Atopy is an objectively measurable trait related to the presence of rhinitis and asthma, but our knowledge about its longitudinal predictors is limited. Data from a 6-year follow-up study of a population sample of children and adolescents (n = 408), aged 7 to 17 years at enrollment, were analyzed to investigate the prevalence and predictors of atopy. METHODS: Case history, including allergic diseases and smoking habits, was obtained by interview and questionnaire. Skin prick test reactivity to common allergens, total serum IgE, airway responsiveness, and pulmonary function were measured using standard techniques. RESULTS: The point prevalence of atopy increased from the first to the second survey, 26% and 44%, respectively; 23% of the participants had a positive skin prick test only at the second survey. Sensitization to house dust mite (HDM) (P < .001), grass (P < .001), dog (P < .001), cat (P < .001), and birch (P = .02) increased significantly in both males and females. No gender differences in the prevalence of positive reactions were found at the first survey, whereas atopy to grass (P = .01) and HDM (P = .02) were more prevalent in males than in females at the second survey. Confining the analysis to participants who were found to be non-atopic at the first survey showed that exposure to maternal smoking (OR 2, CI 1.3 to 3.1; P = .002), increased serum IgE (OR 1.7, CI 1.2 to 2.3; P = .001), new asthma (OR 1.6, CI 1.2 to 2.7; P = .03), and new rhinitis (OR 2.1, CI 1.2 to 3.6; P = .01), but not active smoking, were associated with an increased risk for the presence of a positive skin prick test at the second survey. CONCLUSIONS: This longitudinal population study showed an increase in the point prevalence of atopy in Danish children and adolescents; and, furthermore, that exposure to maternal smoking during childhood, increased serum IgE, and new symptoms of asthma or rhinitis were associated with an increased risk for developing sensitization to common aeroallergens in late adolescence. PMID- 11061473 TI - IgE immune response to Ginkgo biloba pollen. AB - BACKGROUND: The ginkgo (Ginkgo biloba L.) continues to be planted as a shade tree in preference to other species in Seoul, Korea. The proportion of ginkgo to total shade trees was 43.2% in 1998, but the allergenic characteristics of ginkgo pollen has not been elucidated. OBJECTIVES: This study was undertaken to obtain information regarding the skin reactivity rate to ginkgo pollen in a population of Korean subjects with respiratory allergy. Possible ginkgo pollen allergens and the cross-reactivity of ginkgo pollen with other prevalent pollens were also examined. METHODS: Four hundred and forty-seven patients with asthma and/or allergic rhinitis were skin prick tested with extract of ginkgo pollen (1:20 wt/vol). Of these patients, positive skin responders (A/H ratio > or =2+) were selected for ELISA and immunoblot experiments. RESULTS: A total of 21 patients (4.7%) showed skin reactivity (A/H ratio > or =2+) to ginkgo pollen in the skin prick test. They were also cosensitized to many other tree, grass, and weed pollens. Sixteen (76%) of the 21 positive skin responders showed specific IgE responses to ginkgo pollen in ELISA. In inhibitory ELISA, IgE binding to ginkgo pollen was inhibited by more than 80% by oak, ryegrass, mugwort, and ragweed; and 34% by hop Japanese; and 10% by rBet v 2 at 10 microg/mL. In immunoblot, 10 out of 21 sera (48%) reacted to the 15-kD protein of ginkgo pollen, 9 (43%) to 33-35 kD, and 8 (38%) to 36-38 kD. In inhibitory immunoblot, IgE binding to ginkgo pollen proteins was almost completely inhibited by oak, ryegrass, mugwort and ragweed, but only partially by hop Japanese and rBet v 2. CONCLUSION: The skin reactivity rate to ginkgo pollen is approximately 4.7% in a population of Korean subjects with respiratory allergy. Since ginkgo pollen has a high cross reactivity with other prevalent pollens, it could cause clinical symptoms during its pollen season by cross-reacting with the IgE produced in response to other pollens in patients sensitized to multiple pollens. PMID- 11061474 TI - Effect of anti-asthmatic drugs on the response to inhaled endotoxin. AB - BACKGROUND: Endotoxin is a pro-inflammatory agent contaminating the dust that has been associated with the risk to develop pulmonary diseases. There is no data on the protective efficacy of anti-asthmatic drugs on the response induced by inhaled endotoxin in human. METHODS: Twelve mildly asthmatic subjects were submitted weekly to bronchial challenge tests with 20 microg endotoxin. The response was evaluated by the changes in FEV1, blood cells count, neutrophils activation (measured with the luminol-enhanced chemiluminescence) and blood concentration in the acute phase proteins, C-reactive protein (CRP) and haptoglobin. In a double-blind randomized cross-over placebo-controlled design, a single dose each of 500 microg beclomethasone dipropionate, 200 microg salbutamol, and 50 microg salmeterol were administered 30 minutes before the endotoxin challenge test. RESULTS: The 20-microg endotoxin challenge test induced a significant decrease in FEV1 and luminol-enhanced chemiluminescence (P < .001 and <.05, respectively). There was an increase in the blood neutrophils count (P < .05), in CRP (P < .02) and in haptoglobin (P < .03) concentrations. Pretreatment with beclomethasone dipropionate did not have any significant effect on the response to inhaled endotoxin. Salbutamol and salmeterol completely prevent the FEV1 decline due to their potent bronchodilatation activity. Salmeterol and salbutamol did not have any significant effect on the blood inflammation induced by endotoxin inhalation. CONCLUSION: The bronchodilating properties of beta2-agonists prevent the lung function response to inhaled endotoxin. When given in a single dose, an inhaled corticosteroid does not have protective activity on the endotoxin-induced blood inflammation. PMID- 11061475 TI - Case reports of evaluation and desensitization for anti-thymocyte globulin hypersensitivity. AB - BACKGROUND: Biologic products of heterologous sera have been used to treat a variety of conditions. One example is anti-thymocyte globulin (ATG), which is approved for use in the management of renal transplantation and for aplastic anemia. As ATG is a product of heterologous sera it has the potential for adverse reactions, including anaphylaxis. Patients can be skin tested prior to ATG administration to aid in determining hypersensitivity status to ATG. OBJECTIVE: To provide case reports to illustrate evaluation for ATG hypersensitivity. Also, to discuss desensitization procedures for patients who are found to have ATG hypersensitivity, and yet are to receive the medication as it is judged to be essential. CASE REPORTS: We report four patients who were to receive ATG. The results of skin testing and each patient's response to ATG are reviewed to illustrate problems that can occur in evaluating the hypersensitivity status of these patients. Further, some patients also underwent ATG desensitization, but none completed the entire protocol successfully. Their outcomes are reviewed to illustrate problems that can occur with the desensitization procedure. CONCLUSION: Anti-thymocyte globulin is a product of heterologous sera and has the potential to produce anaphylaxis. It is recommended that patients be skin tested prior to administration to aid in determining hypersensitivity status. Those patients who demonstrate hypersensitivity to ATG should not receive ATG unless it is deemed essential and benefits are judged to outweigh risks. In these circumstances, patients are candidates for ATG desensitization. Complications with desensitization occurred in the cases attempted, and highlights that desensitization to ATG, a xenogeneic protein, carries risk and can be difficult. Physicians involved in such cases should be familiar with interpretation of skin tests and problems that can occur with desensitization. PMID- 11061476 TI - Local and systemic reactions during immunotherapy with adsorbed extracts of house dust mite in children. AB - BACKGROUND: We retrospectively evaluated the incidence of local and systemic reactions to injections of adsorbed extracts of house dust mites (Dermatophagoides pteronyssinus and Dermatophagoides farinae) applied according to a conventional schedule in children. METHOD: Eighty-eight patients aged 6 to 15 years, suffering from allergic asthma or asthma together with rhinitis, at various stages of treatment with immunotherapy from January 1989 to November 1997 were included. RESULTS: Out of 5,760 injections, 5,542 (96.21%) were not associated with a reaction, 206 injections (3.57%) caused local reactions (144, <20 mm in diameter; 62, >20 mm), and systemic reactions were seen after 12 injections (0.2%). Twelve patients experienced 12 systemic reactions. Of these, 7 patients (58.3%) experienced no local reactions prior to a systemic reaction. Eleven males and one female had systemic reactions. Most of both local and systemic reactions occurred within less than 30 minutes after the injection. CONCLUSIONS: Our study supports the safety of immunotherapy with house dust mites in children. The majority of reactions were local. Although five of 12 patients who experienced systemic reactions had local reactions prior to a systemic reaction, in general the presence of local reactions was not helpful in predicting which patients would develop systemic reactions. Males and patients with asthma together with rhinitis appeared to be at greater risk for systemic reactions. PMID- 11061477 TI - Relationship between renal dysfunction and bone metabolism disorder in male rats after long-term oral quantitative cadmium administration. AB - To investigate the importance of the cadmium (Cd) exposure condition in the evaluation of toxic effect on renal function and bone metabolism, six groups of Male Wistar rats were given Cd at respective daily doses of 2, 5, 10, 20, 30 and 60 mgCd/kg (as CdCl2) via a gastric tube for 6 consecutive days a week for 60 weeks. In the groups given a low Cd dose (2, 5 and 10 mgCd/kg), relatively more Cd accumulated in the kidney without liver damage than in the liver. In the high Cd dose groups (20, 30 and 60 mgCd/kg), on the other hand, more Cd accumulated in the liver than in the kidney. The daily intake of Cd dose from the intestinal tract in each experimental group was deduced to be about 0.36%-0.54% of the cumulative dose of oral Cd administration. The daily intake of Cd into the body was estimated as 7, 22, 40, 100, 120, 260 microgCd/kg/day in the experimental groups of 2, 5, 10, 20, 30 and 60 mgCd/kg/day, respectively. Increase of plasma enzyme activity (GOT, GPT) and of urinary enzyme excretion (NAG, AAP, GST), reflecting hepatic damage and renal dysfunction, was found in the high Cd dose groups (30 and 60 mgCd/kg) from the 5th week. Non-CdMT concentration in the kidney was also significantly high in the high Cd dose groups. In the low Cd dose groups (2 and 5 mgCd/kg), although the renal Cd concentration was higher than that of the high Cd dose groups, prominent renal dysfunction and hepatic damage were not observed. Regeneration, vacuolization, and eosinophilic bodies in proximal tubular tissue were mainly observed in the groups subjected to 20, 30 and 60 mgCd/kg administration. Very slight regeneration was also observed in the renal proximal tubular tissue at the 30th week for the 5 mgCd/kg and 10 mgCd/kg groups, and at the 60th week for the 2 mgCd/kg group. Remarkable decrease of bone mineral density at the midpoint of the femur was found in the high Cd dose groups. Also, the decrease in bone mineral density was observed before or after the manifestation of the renal dysfunction, depending on the dose and the duration of Cd administration. Urinary excretion of Pyr, DPyr, and Ca increased and plasma BGP decreased in the higher Cd dose groups. Osteoid volume in the femur tissue was not increased significantly by Cd exposure. Based on these results, it was suggested that Cd exposure caused osteoporotic change. The results of the present study suggested that the toxic effect of Cd on renal function and that on bone metabolism were caused at different times and that renal Cd concentration after long-term oral Cd administration depended on the dose and the duration of Cd exposure. PMID- 11061478 TI - Numerical simulations to determine the most appropriate welding and ventilation conditions in small enclosed workspace. AB - In order to improve arc welding work in a small enclosed workspace, numerical simulations were conducted to find the most appropriate welding and ventilation conditions, such as welding currents, hood position and flow rates with no blowhole formation. In the simulations, distributions of airflow vectors and fume concentrations were calculated for two hood opening positions: one faced a welder's breathing zone, the other a contaminant source. As a result it was predicted that a hood opening facing a breathing zone remarkably lowered the fume concentration in the breathing zone compared with that facing a contaminant source. The reliability was confirmed in CO2 arc welding experiments in the enclosed workspace by using a welding robot. In addition, the number of blowholes in welds, examined with x-ray, decreased with the increase in the welding current and with the decrease in the exhaust flow rate. These results showed that the fume concentration near welder's breathing zone and the number of blowholes could be reduced effectively by appropriate selection of the welding current and hood position, and it was confirmed that the numerical simulations were sufficiently useful to predict these appropriate welding conditions. PMID- 11061479 TI - Thermal perception threshold testing for the evaluation of small sensory nerve fiber injury in patients with hand-arm vibration syndrome. AB - The aim of the present study was to investigate whether thermal perception threshold testing is a useful method that could replace pain threshold testing in the evaluation of small sensory nerve fiber injury in vibration-induced neuropathy. Vibration, pain, and thermal (warm and cold) perception thresholds were examined on both middle fingers of 50 patients with hand-arm vibration syndrome and 29 healthy controls of similar age. The patients were divided into three subgroups according to the Stockholm Workshop sensorineural scale. Thermal (warm and cold) thresholds as well as vibration and pain thresholds were significantly more deteriorated among the patients than in the controls. Among the patients, warm thresholds elevated and cold thresholds lowered according to the Stockholm Workshop scale. Thermal thresholds were significantly correlated with pain thresholds, and the sensitivity of the thermal threshold testing tended to be greater than that of the pain threshold testing. The present findings indicate that thermal threshold testing for warm and cold perception can be a useful substitute for pain threshold testing to examine small nerve fiber injury in vibration-induced neuropathy. PMID- 11061480 TI - Zinc protoporphyrin IX concentrations between normal adults and the lead-exposed workers measured by HPLC, spectrofluorometer, and hematofluorometer. AB - To establish the relationship between Zinc protoporphyrin (ZPP) concentrations and blood lead (PbB) levels and to identify reliable analytical methods of ZPP and Protoporhyrin (PP), blood samples were obtained from 263 office workers without the history of occupational lead exposure and 49 lead-acid battery workers. The mean concentrations of PbB for the normal adults and the battery workers were 9.26 microg/dl and 42.60 microg/dl, respectively. The geometric mean concentrations of ZPP and PP by HPLC were 18.73 microg/dl and 2.27 microg/dl for normal adults and were 46.99 microg/dl and 5.53 microg/dl for the exposed workers, respectively. The geometric mean concentrations of ZPP and PP by a spectrofluorometer (SF) were 30.27 microg/dl and 5.16 microg/dl for normal adults and were 50.91 microg/dl and 6.69 +/- 1.39 microg/dl for the exposed workers. The geometric mean ZPP concentration measured by a hematofluorometer (HF) was 30.88 microg/dl for normal adults. The results showed that ZPP concentrations measured by HF were consistently higher than those by HPLC and SF for normal adults, and lower for the exposed workers. ZPP concentrations were not correlated with PbB levels for normal adults but a statistically significant correlation was found among the exposed workers. PMID- 11061481 TI - Performance of respirator filters using quality factor in Korea. AB - A respirator filter of good quality has not only high aerosol collection efficiency but also low air resistance. "Quality factor", which is expressed with aerosol penetration and pressure drop, can be used to rank the performance of respirator filters within the same category. This study focuses on evaluating several respirator filters which are widely used in Korea using quality factor. Two mechanical filters and three filtering facepieces made by different manufacturers were measured aerosol penetrations and pressure drops by an automatic filter tester (CertiTest Model 8110, TSI Inc., St. Paul, USA) at four flow rates of 10, 32, 64 and 85 L/min. NaCl aerosols used were reported to be mean size of 0.1 microm and geometric standard deviation of <1.9. The penetrations and pressure drops of all filters have strong flow rate dependency. The filter quality factors decrease sharply as flow rates are increased. The mechanical filter S and filtering facepiece M are shown better filter quality than others in the same category. Since some certified filters are found to be inappropriate in the workplace exposed to fume, this result suggests that the current certified filter test protocol for respirators should be changed for the new protocol using smaller aerosols. PMID- 11061482 TI - Validity and cost-effectiveness of diagnostic procedures in CS2 poisoning. AB - To determine relatively useful diagnostic procedures of carbon disulfide (CS2) poisoning in terms of validity and cost-effectiveness, several diagnostic tests are evaluated on 1,552 people by prevalence ratio (exposed/nonexposed), dose response relationship, sensitivity and specificity, and the cost of the tests. Several symptoms with high kappa indices are found to be useful in various combinations, showing a consistent dose-response relationship and high exposed nonexposed ratio. In clinicopathologic tests on functions of the kidney, liver and hematopoietic systems, eight items out of 22 have been shown to have significant dose-response relationship, mostly in liver function tests. A thorough dental examination failed to identify any useful indicator peculiar to the CS2 poisoning. Unlike Western people, the prevalence of coronary heart disease among Koreans was too low to be useful in the diagnosis of CS2 poisoning. Among four elective tests, i.e., Minnesota Multiphasic Personality Inventory (MMPI), Brain magnetic resonance imaging (MRI), nerve conduction velocity (NCV), and fluorescent angiography (FAG), the NCV appeared to be a more sensitive and specific test than the others are. Combinations of the tests improved the probability of diagnosing CS2 poisoning cases when any one test out of four was positive. Addition of other valid tests increased the probability of excluding non-cases. It was concluded that diagnosis of CS2 poisoning could be made validly and inexpensively if the diagnostic tests were carefully chosen step by step. PMID- 11061483 TI - Effectiveness of smoking-cessation intervention in all of the smokers at a worksite in Japan. AB - In Japan, the prevalence of smoking among males and females was 56.1% and 14.2%, respectively, in 1997. Male smoking prevalence was exceedingly high as compared to those in other industrialized countries. We conducted a randomized controlled intervention study on smoking cessation for all smokers in a worksite regardless of their willingness to quit smoking. All of the male smokers in a radiator manufacturing factory (n=263) were randomly allocated to an intervention group (n=132) or a control group (n=131). Subjects in the intervention group received individual counseling by a doctor, and those who signed a Smoking Cessation Declaration underwent a five-month intervention. Subjects in the control group received equivalent delayed intervention for four months. The cessation rate after the original intervention was 12.9% (17/132) and 3.1% (4/131) in the intervention and control groups, respectively (p=0.003). Among those who once succeeded in quitting, 48.6% (18/37) maintained cessation at the long-term survey. Overall, the cessation rate was 8.4% (22/263) and the prevalence of smoking among males significantly decreased from 62.9 to 56.7% (p=0.038). As a conclusion, intervention in all smokers at a worksite regardless of their willingness to quit is effective and impacts the overall smoking rate. PMID- 11061484 TI - Effect of repeated exposure to methanol and toluene vapor on the metabolism of rats. AB - Wistar male rats were repeatedly exposed to methanol and toluene vapors individually and simultaneously by inhalation 6 hours a day, five days a week for 4 weeks. Blood was obtained from the tail of the rats up to 23 hours after the end of 4-week exposure and the methanol and toluene concentrations were measured. Major metabolites of methanol and toluene, that is, formic acid and hippuric acid in urine were measured up to 6 days after the end of 4-week exposure. The biological half time of toluene in blood in the simultaneous exposure group was shorter than that in the toluene exposure group. This tendency was almost the same as that for one-day exposure, although the biological half time of solvents in the rat blood was prolonged. The half times of methanol were also longer than those for one-day exposure. PMID- 11061485 TI - Galactosylated liposomes as carriers for targeting meso-2,3-dimercaptosuccinic acid to cadmium storage sites in cadmium exposed mice. AB - In this study an attempt has been made to examine the efficacy of meso-2,3 dimercaptosuccinic acid (DMSA) using galactosylated liposomes as carriers for mobilization of cadmium from the body of mice preexposed to cadmium chloride (0.005 mmoles/kg intraperitoneally daily for 4 days). Cadmium-exposed mice after a rest period of 8 weeks were administered DMSA intravenously, two injections 15 micromoles/kg with an interval of 48 h, as free form of DMSA, or DMSA encapsulated in liposomes composed of phosphatidyl choline, cholesterol and phosphatidyl ethanolamine (7:2:1; PC-lip-DMSA) or in liposomes to which p aminophenyl galactoside had been anchored (Gal-lip-DMSA). Excretion of cadmium through urine and feces was monitored for 5 days. Thereafter animals were sacrificed, liver, kidneys, spleen and isolated hepatocytes were analysed for cadmium, copper and zinc concentration. Efficacy for cadmium mobilization from the body was found to be in the order Gal-lip-DMSA>PC-lip-DMSA>DMSA. These results show that liposomes can be used as targeted carrier system for chelating agents safely and efficiently as compared to administration of free chelating agent. PMID- 11061486 TI - Occupational exposures of pharmacists and pharmaceutical assistants to 60 Hz magnetic fields. AB - We carried out the study to assess, using field surveys and personal dosimetry, the potential exposure of pharmacists and pharmaceutical assistants to 60 Hz magnetic fields in a medical center of Taiwan. Field surveys were conducted twice in the pharmacy where two workers were randomly selected and solicited to wear personal dosimetry instruments for a full-shift assessment of personal exposure. We used an EMDEX II for on site measurements and did not consider any specific instrument or equipment for health care services as potential sources of magnetic field. The results showed that the average magnetic flux densities for the selected areas were between 0.63 mill-Gauss (mG) and 2.23 mG, while the full shift time-weighted-average exposure for the two selected workers was 4.98 mG and 6.54 mG, respectively. Both inadequate consideration for the field survey of the temporal variability in magnetic flux densities over the workday and that the monitored workers spent almost half of the full-shift working in places outside of the study areas may have contributed to such discrepancy in results between field survey and personal dosimetry. This study suggests that the potential for elevated exposure to 60 Hz magnetic fields in health care settings does exist, and that using job title as a surrogate for magnetic fields exposure classification might entail certain degrees of misclassification. Although limited in its scope and sample size, the study presented here seems to demonstrate the inadequacy of using stationary workplace measurements for the assessment of personal occupational exposure to 60 Hz magnetic fields. PMID- 11061487 TI - Ergonomic guidelines for using notebook personal computers. Technical Committee on Human-Computer Interaction, International Ergonomics Association. AB - In the 1980's, the visual display terminal (VDT) was introduced in workplaces of many countries. Soon thereafter, an upsurge in reported cases of related health problems, such as musculoskeletal disorders and eyestrain, was seen. Recently, the flat panel display or notebook personal computer (PC) became the most remarkable feature in modern workplaces with VDTs and even in homes. A proactive approach must be taken to avert foreseeable ergonomic and occupational health problems from the use of this new technology. Because of its distinct physical and optical characteristics, the ergonomic requirements for notebook PCs in terms of machine layout, workstation design, lighting conditions, among others, should be different from the CRT-based computers. The Japan Ergonomics Society (JES) technical committee came up with a set of guidelines for notebook PC use following exploratory discussions that dwelt on its ergonomic aspects. To keep in stride with this development, the Technical Committee on Human-Computer Interaction under the auspices of the International Ergonomics Association worked towards the international issuance of the guidelines. This paper unveils the result of this collaborative effort. PMID- 11061488 TI - Monitoring osteoporosis therapy with bone densitometry: a vital tool or regression toward mediocrity? PMID- 11061489 TI - Clinical Review 116: Bone mineral density, androgens, and the polycystic ovary: the complex and controversial issue of androgenic influence in female bone. PMID- 11061490 TI - Editorial: somatostatin receptor-based scintigraphy and antitumor treatment--an expanding vista? PMID- 11061491 TI - Human ovarian cancers express somatostatin receptors. AB - Characteristics of receptors for somatostatin (SST) analog RC-160 on 17 surgical specimens of human epithelial ovarian cancer and two human ovarian cancer lines were determined by ligand competition assays. The expression of mRNA for four SST receptor subtypes (sst1, sst2A, sst3 and sst5) was investigated by RT-PCR. Thirteen of 17 specimens (76%) exhibited high affinity binding sites for RC-160 with Kd = 6.55 nmol/L and a Bmax = 575.4 fmol/mg membrane protein. Specific receptors for RC-160 were also found in xenografts of OV-1063 and UCI-107 human ovarian cancer lines. The mRNA for sst1 was detected in 65% of the ovarian cancer specimens, while the incidence of sst2A, sst3 and sst5 was 65%, 41% and 24%, respectively. Both ovarian cancer cell lines also expressed mRNA for these four subtypes. The presence of these SST receptor subtypes in human ovarian cancers allows the use of SST analogs and their radionuclide and cytotoxic derivatives for the diagnosis and treatment of this malignancy. PMID- 11061492 TI - Editorial: cardiovascular disease in primary hyperparathyroidism. PMID- 11061493 TI - Augmentation of central arterial pressure in mild primary hyperparathyroidism. AB - Primary hyperparathyroidism (PHPT) is associated with increased cardiovascular risk, although the mechanisms involved remain unclear. Recent evidence has shown increased pulse pressure to be a powerful predictor of cardiovascular events. As increases in pulse pressure are due largely to arterial stiffening, we measured arterial stiffness in 21 subjects with PHPT (18 women and 3 men; 46-71 yr old) and 21 age- and sex-matched healthy controls using pulse wave analysis, a technique that measures peripheral arterial pressure waveforms and generates corresponding central aortic waveforms. This allows determination of the augmentation of central pressure resulting from wave reflection and augmentation index, a measure of vessel stiffness. Metabolic parameters were also measured. The serum calcium level among PHPT subjects was (mean +/- SD) 2.74+/-0.14 mmol/L. pulse wave analysis showed that both augmentation and the augmentation index were significantly higher in the PHPT group vs. controls [16+/-5 vs. 10+/-4 mm Hg (P < 0.001) and 36+/-9% vs. 25+/-6% (P < 0.001)] despite comparable brachial systolic pressures between groups (136+/-13 vs. 134+/-18 mm Hg). Patients with PHPT had higher fasting serum insulin levels [median (range), 15.8 (7.4-39.4) vs. 11.6 (5.1-23) mU/L; P < 0.05] and triglyceride (1.6+/-0.6 vs. 1.2+/-0.4 mmol/L; P < 0.05), but lower high density lipoprotein cholesterol (1.4+/-0.4 vs. 1.6+/-0.3 mmol/L; P < 0.05). These data indicate that subjects with mild PHPT (calcium, <3.0 mmol/L) have increased arterial stiffness, as evidenced by higher augmentation of central aortic pressures. Enhanced vessel stiffness may arise from a combination of structural and functional vascular changes due to hypercalcemia and/or metabolic abnormalities. Increased vascular stiffness in subjects with PHPT may account in part for the increased cardiovascular risk in this group. PMID- 11061494 TI - Metabolic lessons from the study of young adolescents with polycystic ovary syndrome--is insulin, indeed, the culprit? PMID- 11061495 TI - Sensitization to insulin in adolescent girls to normalize hirsutism, hyperandrogenism, oligomenorrhea, dyslipidemia, and hyperinsulinism after precocious pubarche. AB - Precocious pubarche in girls is often preceded by low weight at birth and followed by hirsutism, ovarian hyperandrogenism, and oligomenorrhea in adolescence, the latter usually being accompanied by dyslipidemia and hyperinsulinism, which are, in turn, two major risk factors for cardiovascular disease in later life. We hypothesized that insulin resistance may be a key pathogenetic factor in this sequence. We tested the hypothesis by assessing the effects of an insulin-sensitizing agent, metformin, given at a daily dose of 1275 mg for 6 months to 10 nonobese adolescent girls (mean age, 16.8 yr; body mass index, 21.9 kg/m2; birth weight, 2.7 kg) with hirsutism, ovarian hyperandrogenism (diagnosis by GnRH agonist test), oligomenorrhea, dyslipidemia, and hyperinsulinemia after precocious pubarche. Before the metformin trial, longitudinal studies in these girls had shown that hyperinsulinism was present at prepubertal diagnosis of precocious pubarche, and that it increased markedly in late puberty or early postmenarche. Metformin treatment was well tolerated and was accompanied by a marked drop in hirsutism score, insulin response to oral glucose tolerance test, free androgen index, and baseline testosterone, androstenedione, dehydroepiandrosterone, and dehydroepiandrosterone sulfate levels (all P < 0.01). During metformin treatment, the LH and 17 hydroxyprogesterone hyperresponses to GnRH agonist were attenuated (P < 0.01); serum triglyceride, total cholesterol, and low density lipoprotein cholesterol levels decreased; and high density lipoprotein cholesterol rose. All girls reported regular menses within 4 months. Withdrawal of metformin treatment was followed, within 3 months, by a consistent reversal toward pretreatment conditions. In conclusion, metformin treatment reduced hyperinsulinemia, hirsutism, and hyperandrogenism; attenuated the LH and 17-hydroxyprogesterone hyperresponses to GnRH agonist; improved the atherogenic lipid profile; and restored eumenorrhea in nonobese adolescent girls with a history of precocious pubarche. These observations corroborate the idea that insulin resistance may indeed be a prime factor underpinning the sequence from reduced fetal growth, through precocious pubarche, to adolescent endocrinopathies that are reminiscent of so-called polycystic ovary syndrome. PMID- 11061496 TI - Are ectopic or abnormal membrane hormone receptors frequently present in adrenal Cushing's syndrome? AB - Twenty consecutive patients with adrenal Cushing's syndrome were studied with an in vivo protocol to determine the prevalence and diversity of the presence of ectopic or abnormal hormone receptors in their adrenal tissues. All six patients with bilateral ACTH-independent macronodular adrenal hyperplasia were found to have one or two abnormal adrenal receptors, including those for gastric inhibitory polypeptide, vasopressin (V1-vasopressin), beta-adrenergic agonists, LH/human CG, or serotonin 5-HT4. The presence of abnormal hormone receptors was found to be less frequently present in unilateral adenomas or carcinomas (3 of 14). The identification of abnormal adrenal hormone receptors can allow new pharmacological therapies of hypercortisolism. We suggest that the clinical screening for the presence of abnormal hormone receptors should be conducted in patients with adrenal Cushing's syndrome and, more particularly, in those with ACTH-independent macronodular adrenal hyperplasia, in the hope of offering medical therapy as an alternative to bilateral adrenalectomy. PMID- 11061497 TI - Early changes in serum N-telopeptide and C-telopeptide cross-linked collagen type 1 predict long-term response to alendronate therapy in elderly women. AB - The aim of this study was to determine whether early changes in serum markers of bone resorption could predict long-term responses in bone mineral density (BMD) after alendronate therapy in elderly women. One hundred and twenty women (mean age, 70 yr) were randomized to alendronate or placebo in this double blind, placebo-controlled clinical trial for 2.5 yr. Outcome measures were hip and spine BMD and biochemical markers of bone resorption, including serum N-telopeptide and C-telopeptide cross-linked collagen type I (NTx and CTx, respectively). Serum NTx and CTx were highly correlated at baseline (r = 0.73; P < 0.001) and remained so throughout the study (range, r = 0.36-0.56; all P < 0.05). After treatment with alendronate, serum NTx decreased 30.4+/-16.0% at 6 months, reaching a nadir of 36.7+/-18.0% by 24 months (P < 0.001). Serum CTx decreased 43.5+/-67.0% at 6 months and continued to decrease to 67.3+/-19.3% at 2.5 yr (P < 0.001). Moreover, decreases in serum NTx and CTx at 6 months were correlated with long-term improvements in vertebral BMD at 2.5 yr in patients receiving alendronate therapy (NTx: r = -0.42; CTx: r = -0.31; both P < 0.05). We conclude that early changes in serum NTx and CTx, markers of bone resorption, predict long-term changes in vertebral BMD in elderly women receiving alendronate therapy and provide a useful tool to assess skeletal health. PMID- 11061498 TI - Bone mineral density increases with vitamin D repletion in patients with coexistent vitamin D insufficiency and primary hyperparathyroidism. AB - Two hundred and twenty-nine consecutive subjects, 202 women and 27 men, referred for evaluation of osteoporosis or low bone mineral density (BMD) had serum measurements of immunoreactive PTH (iPTH) and 25-hydroxyvitamin D (25OHD) performed. Fifteen individuals (mean age +/- SE, 75+/-2.4 yr) had depressed serum 25OHD (<15 pg/mL) and concomitantly elevated (>65 pg/mL) iPTH levels. After successful treatment of vitamin D insufficiency in all subjects, iPTH remained inappropriately high or frankly elevated in 5, describing a 2.2% prevalence rate of coexistent primary hyperparathyroidism and vitamin D insufficiency in our population. Despite persistent primary hyperparathyroidism, normalization of serum 25OHD levels in these 5 subjects increased their BMD at an annual rate of 6.3% and 8.2% in spine and hip, respectively. Our results suggest that coexistent vitamin D insufficiency can obscure the diagnosis of primary hyperparathyroidism and, when treated effectively, can result in substantial short-terms gains in BMD despite persistence of the inappropriate production of PTH. PMID- 11061499 TI - Hyperandrogenemia in human immunodeficiency virus-infected women with the lipodystrophy syndrome. AB - A novel lipodystrophy syndrome characterized by insulin resistance, hypertriglyceridemia, and fat redistribution has recently been described in human immunodeficiency virus (HIV)-infected men and women. Women with the HIV lipodystrophy syndrome exhibit a marked increase in waist-to-hip ratio and truncal adiposity; however, it is unknown whether androgen levels are increased in these patients. In this study, we assessed androgen levels in female patients with clinical lipodystrophy based on evidence of significant fat redistribution in the trunk, extremities, neck and/or face (LIPO: n = 9; age, 35.7+/-1.7 yr; BMI, 24.7+/-0.8 kg/m2) in comparison with age- and BMI-matched nonlipodystrophic HIV-infected females (NONLIPO: n = 14; age, 37.6+/-1.1 yr; BMI, 23.4+/-0.6 kg/m2) and healthy non-HIV-infected control subjects (C: n = 16; age, 35.8+/-0.9 yr; BMI, 23.1+/-0.4 kg/m2). Fasting insulin, lipid levels, virologic parameters, and regional body composition using dual energy x-ray absorptiometry were also assessed. Total testosterone [ LIPO, 33+/-6 ng/dL (1.1+/-0.2 nmol/L); NONLIPO, 17+/-2 ng/dL (0.6+/-0.1 nmol/L); C, 23+/-2 ng/dL (0.8+/-0.1 nmol/L); P < 0.05 LIPO vs. C and LIPO vs. NONLIPO] and free testosterone determined by equilibrium dialysis [LIPO, 4.5+/-0.9 pg/mL (16+/-3 pmol/L); NONLIPO, 1.7+/-0.2 pg/mL (6+/-1 pmol/L); C, 2.4+/-0.2 pg/mL (8+/-1 pmol/L); P < 0.05 LIPO vs. C and LIPO vs. NONLIPO] were increased in the lipodystrophic patients. Sex hormone-binding globulin levels were not significantly different between LIPO and C, but were significantly lower in the LIPO vs. NONLIPO patients (LIPO 84+/-7 vs. NONLIPO 149+/-17 nmol/L, P < 0.05). The LH/FSH ratio was significantly increased in the LIPO group compared with the NONLIPO and C subjects (LIPO, 2.0+/-0.6; NONLIPO, 1.1+/-0.1; C, 0.8+/-0.1; P < 0.05 LIPO vs. NONLIPO and LIPO vs. C). Body fat distribution was significantly different between LIPO and C subjects. Trunk to extremity fat ratio (1.46+/-0.17 vs. 0.75+/-0.05, LIPO vs. C, P < 0.05) was increased and extremity to total fat ratio decreased (0.40+/-0.03 vs. 0.55+/ 0.01, LIPO vs. C, P < 0.05). In contrast, fat distribution was not different in the NONLIPO group vs. control subjects. Among the HIV-infected patients, free testosterone correlated with percent truncal fat (trunk fat/trunk mass) (r = 0.43, P = 0.04). These data suggest that hyperandrogenemia is another potentially important feature of the HIV-lipodystrophy syndrome in women. Additional studies are necessary to determine the clinical significance of increased androgen levels and the relationship of hyperandrogenism to fat redistribution and insulin resistance in this population of patients. PMID- 11061500 TI - Hexosamines regulate leptin production in human subcutaneous adipocytes. AB - The hexosamine biosynthetic pathway has recently been proposed as a mechanism through which cells "sense" nutrient flux to regulate leptin release. This study was undertaken to examine the regulation of leptin production by hexosamines in human adipocytes. Adipose tissue UDP-N-acetylglucosamine, an end product of hexosamine biosynthesis, was elevated 3.2-fold, and ob messenger ribonucleic acid was elevated 2-fold in the sc adipose tissue of 17 obese [body mass index (BMI), 41.3+/-12.0 kg/m2; age, 31+/-5 yr] subjects compared to 14 lean (BMI, 23.4+/-1.6 kg/m2; age, 33+/-11 yr) subjects. Serum leptin was increased 2.7-fold in the obese subjects. A significant positive relationship was found between adipose tissue UDP-N-acetylglucosamine and BMI (Spearman correlation = 0.576; P = 0.0007) and between UDP-N-acetylglucosamine and serum leptin (Spearman correlation = 0.4650; P = 0.0145). Treatment of isolated sc adipocytes with 1 mmol/L glucosamine, an intermediate product in UDP-N-acetylglucosamine biosynthesis, increased leptin release 21.4+/-17.6% (mean +/- SD) over control (P = 0.0365) and 74.5+/-82.8% over control (P = 0.0271) in adipocytes from lean (BMI, 23.2+/-1.6 kg/m2; n = 6) and obese (BMI, 55.4+/-13.0 kg/m2,; n = 9) subjects, respectively, by 48 h of culture. Inhibition of UDP-N-acetylglucosamine biosynthesis with 6 diazo-5-oxo-norleucine reduced glucose-stimulated leptin release from cultured adipocytes 21.8+/-32.4% (P = 0.0395; n = 12) and ob gene expression 19.9+/-18.9% (P = 0.0208; n = 8) by 48 h of treatment. These findings suggest that hexosamine biosynthesis regulates leptin production in human adipose tissue. PMID- 11061501 TI - Relationship between hyperinsulinemia and remnant lipoprotein concentrations in patients with impaired glucose tolerance. AB - This study was performed to explore further the association between insulin resistance and plasma remnant lipoprotein (RLP) concentration. For this purpose we used the sum of the plasma insulin concentrations before and 30, 60, 90, 120, and 180 min after a 75-g oral glucose load (sigmaIRI) as a surrogate measure of insulin resistance in 61 subjects with impaired glucose tolerance. SigmaIRI was determined on 2 occasions, before and 16 weeks after initiation of a diet and exercise program. At baseline, sigmaIRI correlated with the sum of the plasma glucose concentrations in response to the 75-g oral glucose load (r = 0.26; P < 0.04) as well as plasma concentrations of triglyceride (r = 0.21; P = 0.09), RLP cholesterol (r = 0.41; P < 0.001), and RLP-triglyceride (r = 0.46; P < 0.001). In contrast, neither total (r = 0.07) nor high density lipoprotein (HDL) cholesterol (r = 0.04) concentrations correlated with sigmaIRI. SigmaIRI was lower in 42 subjects following life-style intervention, associated with significant (P < 0.005) reductions in sigmaglucose, and fasting glucose, insulin, triglyceride, RLP-cholesterol, and RLP-triglyceride concentrations. However, none of these variables decreased in the 19 subjects whose sigmaIRI did not fall. Finally, the change in sigmaIRI following intervention with diet and exercise was significantly associated with differences in sigmaglucose (r = 0.63; P < 0.001) and fasting glucose (r = 0.26; P < 0.05), insulin (r = 0.79; P < 0.001), triglyceride (r = 0.29; P < 0.03), RLP-cholesterol (r = 0.71; P < 0.001), and RLP triglyceride (r = 0.49; P < 0.001) concentrations. These results demonstrate that variations in concentrations of RLPs are highly correlated with changes in sigmaIRI, consistent with the possibilities that 1) RLP measurements are useful estimates of insulin resistance; and 2) an increase in RLP concentrations may provide the mechanistic link between insulin resistance and coronary heart disease. PMID- 11061502 TI - Sexual dimorphism in the influence of advanced aging on adrenal hormone levels: the Rancho Bernardo Study. AB - In recent years, adrenal function and aging has been the subject of intense interest. This cross-sectional study examines age and gender differences in plasma levels of cortisol, dehydroepiandrosterone (DHEA), DHEA-sulfate (DHEAS), and the molar ratio of cortisol/DHEAS in 50-89-yr-old community-dwelling adults. Plasma hormone levels were assayed in samples obtained between 0730 h and 1100 h from 857 men and 735 nonestrogen-using, postmenopausal women. Hormone levels were stratified by 10-yr age groups and compared by two-factor (gender and age) ANOVA. Overall, age and BMI-adjusted DHEA and DHEAS [collectively DHEA(S)] levels were 40% lower and cortisol levels 10% higher in women than men, resulting in a 1.7 fold higher cortisol/DHEAS molar ratio for women (both, P < 0.001). Cortisol levels increased progressively (20% overall) with age in both men and women (both, P < 0.01). Although DHEA(S) levels declined 60% and the cortisol/DHEAS ratio increased 3-fold across the 40-yr age range for both men and women (all P < 0.001), the pattern of the change differed (all P < 0.01 for interaction). For men, DHEA(S) fell in a curvilinear fashion, with the degree of change decreasing with each decade. In contrast, DHEA(S) levels in women fell 40% from the 50s to 60s, were unvarying from 60-80 yr of age, and declined an additional 18% in the 80s. The cortisol/DHEAS ratio increased in a linear fashion for men, but was flat during the 60-80-yr age range for women. Despite these differences in the effect of aging, levels of DHEA(S) remained lower and cortisol and the cortisol/DHEAS ratio higher, in women than men throughout the 50-89-yr age range. These results were independent of adiposity, smoking, and alcohol consumption. In summary, among older, healthy adults DHEA(S) levels are lower and cortisol levels higher in women than men. The age-related decline in adrenal androgens persists into advanced age for both men and women, but exhibits a sexually dimorphic pattern. In contrast, cortisol levels in men and women show a parallel, linear increase with aging. These findings may have important implications for a host of age related processes that exhibit gender differences, including brain function, bone metabolism, and cardiovascular disease. PMID- 11061503 TI - The desmopressin test in the differential diagnosis between Cushing's disease and pseudo-Cushing states. AB - Differentiating Cushing's disease (CD) from pseudo-Cushing (PC) states may still be difficult in current practice. Because desmopressin (1-deamino-8D-arginine vasopressin, DDAVP), a vasopressin analogue, stimulates ACTH release in patients with CD but not in the majority of normal, obese, and depressed subjects, we investigated its ability to discriminate CD from PC states. One hundred seventy three subjects (76 with active CD, 30 with PC, 36 with simple obesity, and 31 healthy volunteers) were tested with an iv bolus of 10 microg DDAVP. Sixty-one of these subjects also underwent a control study with saline. DDAVP induced marked ACTH and cortisol rises in CD (P < 0.005 vs. saline, for both ACTH and cortisol) but not in PC. A significant ACTH elevation occurred upon DDAVP administration also in normal and obese subjects, but it was much smaller than that observed in patients with CD (P < 0.0001). A peak absolute ACTH increase (> or =6 pmol/L), after DDAVP, allowed us to recognize 66 of 76 patients with CD and 88 of 97 subjects of the other groups. The same criterion correctly identified 18 of 20 patients with mild CD (24-h urinary free cortisol < or = 690 nmol/day) and 29 of 30 PC, resulting in a diagnostic accuracy of 94%, which was definitely higher than that displayed by urinary free cortisol, overnight 1-mg dexamethasone suppression test, and midnight plasma cortisol. In conclusion, the DDAVP test seems to be a useful adjunctive tool for the evaluation of hypercortisolemic patients chiefly because of its ability to differentiate mild CD from PC states. PMID- 11061504 TI - Degradation of endogenous and exogenous gastric inhibitory polypeptide in healthy and in type 2 diabetic subjects as revealed using a new assay for the intact peptide. AB - Gastric inhibitory polypeptide (GIP) is susceptible to degradation, but only recently has dipeptidyl peptidase IV been identified as the enzyme responsible. Most RIAs recognize both intact GIP-(1-42) and the noninsulinotropic N-terminally truncated metabolite, GIP-(3-42), hampering measurement of plasma concentrations. The molecular nature of GIP was examined using high pressure liquid chromatography and a newly developed RIA specific for the intact N-terminus of human GIP. In healthy subjects after a mixed meal, intact GIP (N-terminal RIA) accounted for 37.0+/-2.5% of the total immunoreactivity determined by C-terminal assay. High pressure liquid chromatographic analysis of fasting samples by C terminal assay revealed one major peak (73.8+/-2.9%) coeluting with GIP-(3-42). One hour postprandially, two major peaks were detected, corresponding to GIP-(3 42) and GIP-(1-42) (58.1+/-2.7% and 35.7+/-4.2%, respectively). GIP-(3-42) was not detected by N-terminal assay; the major peak coeluted with intact GIP (86.4+/ 5.8% and 81.3+/-0.9%, 0 and 1 h, respectively). After iv infusion, intact GIP constituted 37.1+/-4.1% and 41.3+/-3.4% of the total immunoreactivity in healthy and type 2 diabetic subjects, respectively. The plasma t1/2 was shorter (P < 0.0001) when determined by N-terminal compared with C-terminal assay (7.3+/-1.0 vs. 16.8+/-1.6 and 5.2+/-0.6 vs. 12.9+/-0.9 min, healthy and diabetic subjects, respectively), and both t1/2 were shorter in the diabetic group (P < 0.05). We conclude that dipeptidyl peptidase IV is important in GIP metabolism in humans in vivo, and that an N-terminally directed assay is required for determination of plasma concentrations of biologically active GIP. PMID- 11061505 TI - Liquid chromatography-mass spectrometry detection of corticotropin-releasing hormone and proopiomelanocortin-derived peptides in human skin. AB - We have previously shown expression of CRH and POMC genes and peptides in the human skin. To ascertain the identity of those peptides, we used methods of peptide extraction and purification combined with the highly specific technique of liquid chromatography-mass spectrometry. Testing extracts of human skin, we identified endogenous peptides with masses and retention times corresponding to CRH, ACTH 1-39, ACTH 1-13, and alpha-MSH standards. Thus, conclusive evidence is provided for the presence of CRH and the POMC-derived ACTH 1-39, ACTH 1-13, and alpha-MSH peptides in human skin. Direct identification of these peptides is consistent with translation of the corresponding genes, and it also suggests intermediate pituitary lobe-like POMC peptide processing. PMID- 11061506 TI - The A19G polymorphism in the 5' untranslated region of the human obese gene does not affect leptin levels in severely obese patients. AB - Recently, the presence of different polymorphisms in the regulatory region of the ob gene has been associated with variations in leptin levels. However, the results of these studies are still contradictory. The aim of the present investigation was to evaluate the presence of the A19G polymorphism in an Italian population of obese patients and to verify its association with leptin levels and anthropometric, metabolic, and clinical parameters. Two hundred five obese patients [body mass index (BMI) > 36 kg/m2; 135 women and 70 men; mean age, 46.9+/-14.23 yr] were screened for presence of the polymorphism; 61 normal-weight controls (mean BMI, 21.05 kg/m2; 53 women, 8 men) were also screened to compare polymorphism frequency. For obese patients, BMI, waist-to-hip ratio, resting energy expenditure, body composition, fasting leptin, total cholesterol, high density lipoproteins, triglycerides, and caloric intake were determined. Genotype frequencies in obese and control subjects were compared using the contingency table chi-square test; in obese subjects an ANOVA was performed to evaluate association between the polymorphism and several clinical parameters. No significant differences in genotype distribution between control and obese subjects were found. No significant correlations were found between this polymorphism and serum leptin levels and the other parameters considered. These findings confirm the results obtained in both a Finnish and a French population; taken together, these observations might rule out a significant role for the A19 >G polymorphism in the regulation of leptin levels and other clinical, anthropometric, and metabolic parameters. PMID- 11061507 TI - Administration of a single low dose of recombinant human thyrotropin significantly enhances thyroid radioiodide uptake in nontoxic nodular goiter. AB - Radioiodine (131I) is increasingly used as treatment for volume reduction of nontoxic, nodular goiter. A high dose of 131I is often needed because of low thyroid radioiodide uptake (RAIU). We investigated whether pretreatment with a single, low dose of recombinant human TSH (rhTSH; Thyrogen, Genzyme Transgenics Corp.) enhances RAIU in 15 patients with nontoxic, nodular goiter (14 women and 1 man; aged 61+/-11 yr). Four patients were studied twice, and 1 patient was studied 3 times. RAIU was measured both under basal conditions and after pretreatment (im) with rhTSH, given either 2 h (0.01 mg; n = 7) or 24 h [0.01 mg (n = 7) or 0.03 mg (n = 7)] before 131I administration (20-40 microCi). Serum levels of TSH, free T4 (FT4), and total T3 were measured at 2, 5, 8, 24, 48, 72, 96, and 192 h after rhTSH administration. After administration of 0.01 mg rhTSH, serum TSH rose from 0.7+/-0.5 to a peaklevel of 4.4+/-1.1 mU/L (P < 0.0001), FT4 rose from 16.0+/-2.6 to 18.5+/-3.7 pmol/L (P < 0.0001), and T3 rose from 2.10+/ 0.41 to 2.63 - 0.66 nmol/L (P < 0.0001). After administration of 0.03 mg rhTSH, TSH rose from 0.6+/-0.4 to 15.8+/-2.3 mU/L (P < 0.0001), FT4 rose from 15.2+/-1.5 to 21.7+/-2.9 pmol/L (P < 0.0001), and T3 rose from 1.90+/-0.43 to 3.19+/-0.61 nmol/L (P < 0.0001). Peak TSH levels were reached at 5-8 h and peak FT4 and T3 levels at 8-96 h after rhTSH administration. Administration of 0.01 mg rhTSH 2 h before 131I increased 24-h RAIU from 30+/-11% to 42+/-10% (P < 0.02), 0.01 mg rhTSH administered 24 h before 131I increased 24-h RAIU from 29+/-10% to 51+/-10% (P < 0.0001), and 0.03 mg rhTSH administered 24 h before 131I increased 24-h RAIU from 33+/-11% to 63+/-9% (P < 0.0001). After administration of 0.01 mg rhTSH 2 h before 131I, 24-h RAIU did not increase in 1 patient, whereas the increase in 24 h RAIU was less than 10% in 2 other patients. In contrast, administration of rhTSH 24 h before 131I increased 24-h RAIU by more than 10% in all 14 patients (by >20% in 10 and by >30% in 6). In conclusion, pretreatment with a single, low dose of rhTSH in patients with nontoxic, nodular goiter increased RAIU considerably. Our observations hold promise that administration of rhTSH before 131I therapy for nontoxic, nodular goiter will allow treatment with lower doses of 131I in these patients. PMID- 11061508 TI - Effects of sleep and sleep deprivation on interleukin-6, growth hormone, cortisol, and melatonin levels in humans. AB - The objective of this study was to evaluate the effects of nocturnal sleep, partial night sleep deprivation, and sleep stages on circulating concentrations of interleukin-6 (IL-6) in relation to the secretory profiles of GH, cortisol, and melatonin. In 31 healthy male volunteers, blood samples were obtained every 30 min during 2 nights: uninterrupted, baseline sleep and partial sleep deprivation-early night (awake until 0300 h). Sleep was measured by electroencephalogram polysomnography. Sleep onset was associated with an increase in serum levels of IL-6 (P < 0.05) during baseline sleep. During PSD-E, the nocturnal increase in IL-6 was delayed until sleep at 0300 h. Sleep stage analyses indicated that the nocturnal increase in IL-6 occurred in association with stage 1-2 sleep and rapid eye movement sleep, but levels during slow wave sleep were not different from those while awake. The profile of GH across the 2 nights was similar to that of IL-6, whereas the circadian-driven hormones cortisol and melatonin showed no concordance with sleep. Loss of sleep may serve to decrease nocturnal IL-6 levels, with effects on the integrity of immune system functioning. Alternatively, given the association between sleep stages and IL-6 levels, depressed or aged populations who show increased amounts of REM sleep and a relative loss of slow wave sleep may have elevated nocturnal concentrations of IL-6 with implications for inflammatory disease risk. PMID- 11061509 TI - Arginine counteracts the inhibitory effect of recombinant human insulin-like growth factor I on the somatotroph responsiveness to growth hormone-releasing hormone in humans. AB - Insulin-like growth factor I (IGF-I) exerts a negative feedback effect on GH secretion via either direct actions at the pituitary level or indirect ones at the hypothalamic level, through stimulation of somatostatin (SS) and/or inhibition of GHRH release. In fact, recombinant human IGF-I (rhIGF-I) in humans inhibits spontaneous GH secretion as well as the GH response to GHRH and even more to GH/GH-releasing peptides, whose main action is on the hypothalamus, antagonizing SS and enhancing GHRH activity. The aim of the present study was to further clarify in humans the mechanisms underlying IGF-I-induced inhibition of somatotroph secretion. In six normal young volunteers (all women; mean +/- SEM: age, 28.3+/-1.2 yr; body mass index, 21.3+/-1.2 kg/m2) we studied the GH response to GHRH (1 microg/kg, iv, at 0 min), both alone and combined with arginine (ARG; 0.5 g/kg, iv, from 0-30 min), which probably acts via inhibition of hypothalamic SS release, after pretreatment with rhIGF-I (20 microg/kg, sc, at -180 min) or placebo. rhIGF-I increased circulating IGF-I levels (peak at -60 vs. -180 min: 54.9+/-3.9 vs. 35.9+/-3.3 mmol/L; P < 0.05) to a reproducible extent, and these levels remained stable and within the normal range until 90 min. The mean GH concentration over 3 h (from -180 to 0 min) before ARG and/or GHRH was not modified by placebo or rhIGF-I. After placebo, the GH response to GHRH (peak, 23.6+/-2.9 microg/L) was strikingly enhanced (P < 0.05) by ARG coadministration (69.6+/-9.9 microg/L). rhIGF-I blunted the GH response to GHRH (13.1+/-4.5 microg/L; P < 0.05), whereas that to GHRH plus ARG was not modified (59.5+/-8.9 microg/L), although it occurred with some delay. Mean glucose and insulin concentrations were not modified by either placebo or rhIGF-I. In conclusion, ARG counteracts the inhibitory effect of rhIGF-I on somatotroph responsiveness to GHRH in humans. These findings suggest that the acute inhibitory effect of rhIGF I on the GH response to GHRH takes place on the hypothalamus, possibly via enhancement of SS release, and that ARG overrides this action. PMID- 11061510 TI - Search for abnormalities of nuclear corepressors, coactivators, and a coregulator in families with resistance to thyroid hormone without mutations in thyroid hormone receptor beta or alpha genes. AB - The syndrome of resistance to thyroid hormone (RTH) is characterized by decreased tissue responsiveness to thyroid hormones. Inheritance is usually autosomal dominant due to mutations in the ligand-binding domain or adjacent hinge region of the thyroid hormone receptor beta (TRbeta) gene. Six of 65 families with the RTH phenotype studied in our laboratory had normal TRbeta1 and TRbeta2 gene sequences. Their clinical characteristics were not different from those of subjects with TRbeta gene mutations. Four of the 6 families were amenable to linkage analysis, and TRalpha involvement was excluded. Candidate genes were then evaluated for their possible involvement in the RTH phenotype in these 4 families: 2 coactivators [NCoA-1 (SRC-1) and NCoA-3 (AIB-1)], 2 corepressors (NCoR and SMRT), and a coregulator (RXRgamma). DNA was obtained from 8 affected subjects and 41 of 45 living first degree relatives. In 2 of the 4 families, the mode of inheritance could be determined by pedigree analysis and was found to be autosomal dominant. Linkage analyses were performed using polymorphic markers near or within the 5 candidate genes. When analyses were not informative or linkage could not be excluded, direct sequencing of the genes in question was performed. Involvement of NCoA-1 was excluded in all four families assuming autosomal dominant inheritance. Roles for NCoR, SMRT, and NCoA-3 were excluded in three and a role for RXRgamma was excluded in two of the four families. However, if the two families without proven dominant mode of inheritance were compound heterozygous, only the involvement of NCoA-1 could be excluded in both. Roles for NCoR, SMRT, and RXRgamma were excluded in one of these two families. Thus, NCoA-1 and RXRgamma genes were not found to be the cause of RTH in subjects without TR gene mutations even though the absence of NCoA-1 and RXRgamma is the cause of RTH in mice. Involvement of other candidate genes in the mediation of thyroid hormone action as well as intracellular hormone transport needs to be explored in these families with non-TRbeta, TRalpha RTH. PMID- 11061511 TI - Melatonin does not shift circadian phase in baboons. AB - It has been suggested that the pineal hormone melatonin can modulate circadian rhythmicity and may have clinical utility in treating biological clock disorders. Thus, there is considerable clinical interest in using melatonin to treat disorders, such as jet-lag. Yet, despite growing enthusiasm for the use of melatonin, it is not clear whether melatonin indeed shifts the circadian phase in humans and other primate species. Thus, to assess whether melatonin can influence circadian phase, we studied the phase-shifting effects of melatonin on baboons to provide insights into the role of melatonin. Mel- atonin was administered orally to baboons (0.5, 3, 5, or 10 mg) either in the early morning hours from circadian time (CT) 0 to CT3 or late in the afternoon from CT9 to CT12, and changes in circadian phase were assessed. Surprisingly, at all doses and times tested, melatonin did not shift circadian phase. Physical activity was reduced after 5- and 10-mg doses given late in the afternoon, but not after doses given early in the morning. These observations suggest that melatonin does not shift circadian phase in baboons using doses similar to those prescribed for treating human circadian system disorders. PMID- 11061512 TI - Detection of recurrent thyroid cancer by sensitive nested reverse transcription polymerase chain reaction of thyroglobulin and sodium/iodide symporter messenger ribonucleic acid transcripts in peripheral blood. AB - To investigate whether circulating thyroglobulin (Tg) messenger ribonucleic acid (mRNA) and sodium/iodide symporter (NIS) mRNA transcripts in peripheral blood are valuable in the follow-up of patients with thyroid cancer, we developed highly sensitive nested Tg and NIS mRNA detection assays and compared their accuracy with serum thyroglobulin (sTg) and whole body scan with 131I during the monitoring of 34 patients with well differentiated thyroid carcinoma who had undergone total thyroidectomy (17 of 34 also submitted to thyroid ablation with radioiodine) and were taking T4. Circulating Tg mRNA was found in 13 of 34 patients, 5 of 13 with detectable and 8 of 13 with undetectable sTg. From these 8 patients with undetectable Tg, 6 showed no cervical radioiodine uptake, and 3 presented proven metastatic disease (2 of them positive for antithyroglobulin antibodies). NIS mRNA was detected in 11 of 34 patients, but its measurement did not improve the ability to detect patients with metastases. Overall, identification of metastatic thyroid cancer was better associated with Tg mRNA than with NIS mRNA, sTg, or whole body scan (83% vs. 16.6% vs. 50% vs. 50%; P < 0.001). These data showed that circulating Tg mRNA is not only a more sensitive marker of residual thyroid tissue or thyroid cancer than sTg, particularly in patients during T4 therapy and with positive antithyroglobulin antibodies, but also was more sensitive than NIS mRNA in all patients. PMID- 11061513 TI - Natural history of a proinsulin-secreting insulinoma: from symptomatic hypoglycemia to clinical diabetes. PMID- 11061514 TI - Circulating thyrotropin bioactivity in sporadic central hypothyroidism. AB - The etiopathogenesis of sporadic central hypothyroidism (CH) involves pituitary and hypothalamic lesions. Pituitary CH (pCH) implies a diminished number of functioning thyrotropes, accounting for the quantitative impairment of TSH secretion. Hypothalamic CH (hCH) is characterized by normal or even increased TSH concentrations and qualitative abnormalities of TSH secretion, including a decreased bioactivity of circulating TSH. However, controversy still exists about the actual occurrence of bioinactive TSH among CH patients, and no data are available in pCH. Therefore, we studied 41 CH patients with different hypothalamic-pituitary disorders. Immunoreactive TSH (TSH-I) ranged from 0.08 11.1 mU/L (normal, 0.24-4.0), free T4 (FT4) ranged from 0.6-8.8 pmol/L (normal, 9 18), and FT3 ranged from 1.2-5.4 pmol/L (normal, 4-8). A blunted TSH response to TRH (<4 mU/L), indicating prevalent pCH, was found in 56% of the patients, and a net TSH-I increment > or =4 mU/L, indicating prevalent hCH, was found in the remaining 44%. Net TSH-I increments showed significant correlation with basal FT4 (P < 0.02), indicating the relevance of pituitary TSH reserve in the pathogenesis of CH. Circulating TSH was immunoconcentrated and tested in bioassay and in ricin affinity chromatography. The ratio between biological (B) and immunological (I) activities of circulating TSH was reduced (n = 25; TSH B/I, 0.38+/-0.19) compared to the values recorded in normal subjects (n = 26; TSH B/I, 1.53+/-0.54; P < 0.001) and primary hypothyroid patients (n = 24; TSH B/I, 0.74+/-0.31; P < 0.001), but no difference between pCH (n = 9; 0.36+/-0.16) and hCH (n = 16; 0.39+/-0.20) was seen. TSH B/I values in CH patients showed a limited overlap with normal values (20%) and a highly significant correlation with the FT3 response to endogenous TRH-stimulated TSH (P < 0.005). The elevated sialylation degree of TSH molecules may explain part of these findings. In conclusion, the secretion of TSH molecules with reduced bioactivity is a common alteration in the patients with hypothalamic-pituitary lesions, contributing along with the impairment of pituitary TSH reserve to the pathogenesis of CH. PMID- 11061515 TI - Mutations in the steroidogenic acute regulatory protein (StAR) in six patients with congenital lipoid adrenal hyperplasia. AB - Congenital lipoid adrenal hyperplasia (lipoid CAH), the most severe form of CAH, is caused by mutations in the steroidogenic acute regulatory protein (StAR). Lipoid CAH is common among the Japanese, Korean, and Palestinian Arab populations, but is rare elsewhere. We describe six patients with lipoid CAH: four Japanese, one Palestinian, and one Guatemalan Native American. All had classical clinical presentations of normal female external genitalia in both genetic sexes, with severe glucocorticoid and mineralocorticoid deficiency presenting in the first month of life. Quite atypically, one patient had small adrenal glands shown by computed tomographic scanning. The StAR genes were characterized in all six patients. Three of the Japanese patients were compound heterozygotes for the common Japanese mutation Q258X in association with three different novel frameshift mutations; the fourth Japanese patient was homozygous for the mutation R182L, which is common among Palestinian patients but has not been described previously in a Japanese patient. Our Palestinian and Native American patients were each homozygous for novel frameshift mutations. Thus we have found five new frameshift mutations, but no new amino acid replacement (missense) mutations. This would be consistent with the view that only a small number of residues in the StAR protein are crucial for biological activity. The tomographic finding of small adrenals in a patient with genetically proven lipoid CAH due to a StAR mutation suggests a substantially broader spectrum of clinical findings in this disease than has been appreciated previously. PMID- 11061516 TI - Use of recombinant human thyrotropin before radioiodine therapy in patients with advanced differentiated thyroid carcinoma. AB - The use of 131I for radioablative therapy in patients with differentiated thyroid cancer (DTC) requires a sufficient serum concentration of TSH for efficient thyroid tissue uptake of iodine. We describe the use of recombinant human TSH (rhTSH) in conjunction with ablative radioiodine therapy (RIT) in 11 patients (16 total treatments) with advanced and/or recurrent DTC (5 papillary, 6 follicular) for whom withdrawal of thyroid hormone suppression therapy (THST), the standard method to increase serum TSH, was not an option. Indications for rhTSH use in these patients included inability to tolerate withdrawal of thyroid hormones due to very poor physical condition or inability to achieve sufficient serum TSH levels after THST withdrawal. Ten patients had undergone thyroidectomy, and most (9 of 11) had received prior ablative RIT after THST withdrawal. Baseline thyroglobulin levels ranged from 25 to nearly 30,000 ng/mL, reflecting the heterogeneity of the patient population. In 7 cases (5 patients), posttherapy thyroglobulin levels assessed at a mean of 4.3 months (range, 2-10 months) after 131I therapy were decreased by at least 30% compared to pretherapy levels. In follow-up visits, an additional 3 patients showed marked clinical improvement or decreased or stabilized tumor burden in whole body scans compared to pretherapy scans. Three patients died of progressive disease within 2 months of therapy before follow-up assessments occurred. No adverse events were reported among the 8 surviving patients. The results suggest that rhTSH offers a promising alternative to THST withdrawal to allow ablative RIT after effective TSH stimulation in patients with advanced recurrent DTC who would not otherwise be able to receive this treatment. This therapeutic indication extends the clinical potential of this new agent, already demonstrated to be effective for use with 131I for diagnostic purposes. PMID- 11061517 TI - Chronic alcohol intake differently influences glucose metabolism according to nutritional status. AB - We investigated the potential different effects of a chronic alcohol intake on glucose metabolism according to nutritional status in growing rats. Eighty weanling 4-week-old male Sprague Dawley rats were fed with low (5%, wt/wt) or control (22%) protein diet for 8 weeks. Each group was subdivided into alcohol (5 g/kg(-1) x day(-1)) or saline gavage rats during the last 4 weeks. At 12 weeks of age, we measured the weights of the body, pancreas, and epididymal fat; glycogen synthase activity of gastrocnemius muscle; and insulin content of the pancreas. We performed an ip glucose tolerance test and a euglycemic hyperinsulinemic clamp test. Weight gain was almost arrested in protein-deficient rats. The relative weight and insulin content of the pancreas and glycogen synthase activity were not different among the four groups, but the relative amount of epididymal fat decreased only in protein-deficient saline rats. Insulin response after glucose challenge and glucose disposal rate during the euglycemic clamp were both markedly decreased in protein-deficient saline rats, but not changed in protein deficient alcohol rats. Protein-deficiency per se causes deterioration both in insulin secretory function and in sensitivity, but these defects are protected by a chronic alcohol intake. In a protein-sufficient state, alcohol intake gives no additional effects on glucose metabolism. Therefore, according to individual nutritional status, the metabolic effect of alcohol intake appears differently. PMID- 11061518 TI - High dose recombinant human growth hormone (GH) treatment of GH-deficient patients in puberty increases near-final height: a randomized, multicenter trial. Genentech, Inc., Cooperative Study Group. AB - GH production rates markedly increase during human puberty, mostly as an amplitude-modulated phenomenon. However, GH-deficient children have been dosed on a standard per kg BW basis similar to prepubertal children. This randomized study was designed to compare the efficacy and safety of standard recombinant human GH (rhGH) therapy (group I, 0.3 mg/kg x week) vs. high dose therapy (group II, 0.7 mg/kg x week) in GH-deficient adolescents previously treated with rhGH for at least 6 months. Ninety-seven children with documented evidence of GH deficiency (peak GH in response to stimuli, <10 ng/mL), with either organic or idiopathic pathology, were recruited. Both groups were matched for sex (group I, 42 males and 7 females; group II, 41 males and 7 females), age [group I, 14.0+/-1.6 (+/ SD) yr; group II, 13.7+/-1.6], standardized height (group I, -1.4+/-1.1; group II, -1.2+/-1.1), bone age (group I, 13.1+/-1.3 yr; group II, 13.1+/-1.3) etiology, maximum stimulated GH, previous growth rate, and midparental target height. All subjects were in puberty (Tanner stage 2-5) at study entry. Of the 97 subjects enrolled, 45 were treated for 3 yr or more; 48 completed the study. Of the subjects who discontinued the study, the most common reason was satisfaction with their height, although others discontinued for adverse events or personal reasons. The frequency of patients who discontinued was the same in both groups. The primary efficacy analysis was the difference between dose groups for near adult height, defined as the height attained at a bone age of 16 yr or more in males and 14 yr or more in girls; all subjects who qualified were included in the analysis. This difference was statistically significant at 4.6 cm by analysis of covariance (ANCOVA; P < 0.001; n = 75). For subjects who received at least 4 yr of rhGH treatment, the difference between dose groups at that time point was 5.7 cm (by ANCOVA, P = 0.024; n = 20). The mean height SD score at near-adult height was -0.7+/-0.9 in the standard dose group and 0.0+/-1.2 in the high dose group. At 36 months the cumulative change in height (centimeters) was 21.5+/-5.3 cm (group I) vs. 25.1+/-4.9 (group II; P < 0.001, by ANCOVA); the change in Bayley Pinneau predicted adult height was 4.8+/-4.2 cm (group I) vs. 8.4+/-5.7 (group II; P = 0.032). Median plasma IGF-I concentrations at baseline were 427 microg/L (range, 204-649) in group I and 435 microg/L (range, 104-837) in group II; at 36 months they were 651 microg/L (range, 139-1079) in group I vs. 910 microg/L (range, 251-1843) in group II (P = NS). No difference in change in bone age was detected between groups at any interval. High dose rhGH was well tolerated, with a similar safety profile as standard dose treatment and no difference in hemoglobin A1c or glucose concentrations between groups. In summary, compared to conventional treatment, high dose rhGH therapy in adolescents 1) increased near adult height and height SD scores significantly, 2) did not increase the rate of skeletal maturation, and 3) appears to be well tolerated and safe. In conclusion, high dose rhGH therapy may have a beneficial effect in adolescent GH-deficient patients, particularly those who are most growth retarded at the start of puberty. PMID- 11061519 TI - Relation of plasma oxytocin and prolactin concentrations to milk production in mothers of preterm infants: influence of stress. AB - Responses of oxytocin and PRL to mechanical breast pumping and the influence of physiological indicators of stress were measured at 2, 4, and 6 weeks postpartum to determine potential causes of inadequate milk production in 18 women with prematurely delivered, nonnursing (<1500 g) infants. Median milk production was similar to that reported in breastfeeding mothers, but a third of mothers were producing less than half as much by week 6. Plasma oxytocin was similar to that previously reported for breastfeeding mothers. The oxytocin area under the curve (AUC) for breast-pumping sessions (70 min) was correlated at each occasion (r = 0.37, 0.58, and 0.55, respectively) with milk yield. Unlike reports of PRL levels in breast-feeding women, PRL AUC declined between weeks 2 and 6 weeks postpartum (P = 0.03); significant increases in plasma PRL occurred in response to pumping at 2 and 4 weeks, but not at 6 weeks. Salivary amylase, a measure of alpha adrenergic activity, was highly negatively correlated on each occasion with PRL AUC (r = -0.58, -0.68, and -0.86, respectively), but not with oxytocin. Salivary cortisol was negatively correlated to a lesser degree. We hypothesize that deficiencies in preterm lactation are mediated in part upon stress-induced suppression of PRL secretion through an adrenergic mechanism. PMID- 11061520 TI - 3beta-hydroxysteroid dehydrogenase/delta5-->4-isomerase activity associated with the human 17beta-hydroxysteroid dehydrogenase type 2 isoform. AB - The type 2 isoform of human 17beta-hydroxysteroid dehydrogenase (17betaHSD2) efficiently catalyzes the oxidative metabolism of androgens and estrogens, and it is expressed in a large series of human peripheral tissues. To obtain a better understanding of the regulation of local steroid biosynthesis and metabolism in human tissues, we have established a dual steroidogenic activity of the 17betaHSD2 enzyme after transfection of human 17betaHSD2-transfected human embryonic kidney (293) cells. After transient transfection, the metabolism of testosterone, pregnenolone, and dehydroepiandrosterone (DHEA) in intact transfected 293 cells was evaluated by TLC-based radiometric assays. 17betaHSD2 transfected cells converted 91% of testosterone (1 micromol/L) into androstenedione in a 2-h incubation period. In addition, pregnenolone (1 micromol/L) was converted to progesterone (18.5%), whereas DHEA (1 micromol/L) was metabolized to androstenedione (8.3% conversion) in a 15-h incubation period. The kinetics of the 3beta-hydroxysteroid dehydrogenase (3betaHSD) and 17betaHSD2 activities using cell homogenate protein of stably transfected 293 cells indicated that the catalytic efficiency (apparent catalytic efficiency = maximum velocity/Km) of this 3betaHSD activity is approximately 2000-fold (pregnenolone as substrate) or 3000-fold (DHEA as substrate) weaker than that of 17betaHSD2 activity. It is noteworthy, however, that the apparent catalytic efficiency of the HSD3B2 gene product is only approximately 50-fold higher than that of the 3betaHSD aspect of the 17betaHSD2 gene product. Pregnenolone or DHEA effectively inhibited 17betaHSD2 activity in a noncompetitive fashion. Furthermore, the potent 5alpha-reductase/3betaHSD inhibitor, 17beta-N,N-diethylcarbamoyl-4-methyl 4-aza-5alpha-androstane-3-one , inhibited neither 3betaHSD nor 17betaHSD2 activities. We conclude that human 17betaHSD2 enzyme exhibits 3betaHSD activity. Notwithstanding that this 3betaHSD activity is reduced compared to 17betaHSD oxidative activity, it may account for at least some of the reports of 3betaHSD activity found in human peripheral tissues that express notable amounts of the 17betaHSD2 isozyme as well as in individuals with severe classic 3betaHSD deficiency. PMID- 11061521 TI - Effect of weight loss on the pulsatile insulin secretion. AB - The aim of the study was to assess whether pulsatile insulin secretion is variable in the same individual and is related to changes in insulin sensitivity. Insulin sensitivity and pulsatility were measured before and after weight reduction in nine obese subjects. A pulsatility analysis was done using the PulsDetekt program. Blood was sampled every 2 min over a period of 90 min. The secretion randomness was quantified using approximate entropy (ApEn), and ApEn normalized by SD of the insulin time series (nApEn). Lower values indicate more regular secretion. Insulin sensitivity was measured using the homeostasis model assessment. Data are presented as median, minimum-maximum. After weight loss insulin sensitivity was increased (12.16, 7.60-76.70 vs. 38.96, 19.88-74.96%), the number of insulin pulses was reduced (11, 8-16 vs. 9,6-12), and they were more regular (ApEn, 0.92, 0.53-133 vs. 0.69,0,40-1.27; nApEn, 1.07, 0.74-1.33 vs. 0.97, 0.54-1.42). Before and after the weight loss there was a correlation between ApEn and nApEn and insulin sensitivity. Therefore, insulin secretion regularity is variable in the same individual and is related to insulin sensitivity. PMID- 11061522 TI - Thyrotoxicosis in prepubertal children compared with pubertal and postpubertal patients. AB - The course of Graves' thyrotoxicosis in 7 prepubertal children (6.4+/-2.4 yr) was compared with that in 21 pubertal (12.5+/-1.1 yr) and 12 postpubertal (16.2+/ 0.84 yr) patients. In the prepubertal group the main complaints were weight loss and frequent bowel movements (86%), whereas typical symptoms (irritability, palpitations, heat intolerance, and neck lump) occurred significantly less often (P < 0.01). The most prominent manifestation at diagnosis was accelerated growth and bone maturation: their height SD score was significantly greater than that of the pubertal and postpubertal patients (2.6+/-0.7 us. 0.15+/-0.65 and 0.15+/-0.9, respectively, P < 0.001), and their bone age to chronological age ratio was 1.39+/-0.35 compared with 0.98+/-0.06 in the pubertal children (P = 0.02). T3 levels were also significantly higher than in the other two groups (9.9+/-2.9 nmol/L vs. 6.32+/-1.9 nmol/L and 6.02+/-2.0 nmol/L, P = 0.01). All patients were initially prescribed antithyroid drugs (ATDs). Overall, adverse reactions to ATDs occurred in 35%, with a higher rate among the prepubertal children (71%) than the pubertal (28%) and postpubertal (25%) patients (P = 0.08). Major adverse reactions were noted in two children, both prepubertal. Remission was achieved in 10 patients (28%). Although the rate of remission did not differ among the three groups, time to remission tended to be longer in the prepubertal children (P = 0.09). In conclusion, thyrotoxicosis has an atypical presentation and more severe course in prepubertal children. Considering their adverse reactions to ATD, overall low remission rate, and long period to remission, definitive treatment should be considered earlier in this age group. PMID- 11061523 TI - Effect of systemic oxytocin administration on dexamethasone-induced leptin secretion in normal and obese men. AB - To establish whether the regulatory mechanism of leptin secretion is sensitive to oxytocin (OT), seven healthy nonobese men were tested with dexamethasone (dex; 4 mg, iv, at 0730 h) in feeding (2000 Cal given at 3 meals over 7 h) conditions either in the absence (iv normal saline infusion) or in the presence of a constant iv infusion of OT (1, 2, or 4 mIU/min from 0730 h for 10 h). In six additional subjects under similar experimental conditions, normal saline or OT (1, 2, or 4 mIU/min from 0730 h for 10 h) were infused iv without the previous treatment with dexamethasone. Serum leptin concentrations were measured in samples taken at 60-min intervals during infusion. Leptin levels remained constant during the infusion of normal saline or OT (1, 2, or 4 mIU/min) alone. In contrast, serum leptin concentrations rose significantly from the baseline after dex administration. The leptin response to dex was not modified by the concomitant infusion of 1 mIU/min OT, whereas it was completely abolished by the administration of 2 or 4 mIU/min OT. These findings led us to evaluate the secretory pattern of leptin in 12 obese patients in similar experimental conditions. In all patients basal leptin levels were significantly higher than those in normal weight subjects. In 6 obese subjects, the infusion of OT alone (1, 2, or 4 mIU/min) was unable to change serum leptin levels. In the remaining 6 obese subjects, dex administration significantly increased serum leptin levels; however, the leptin response to dex was not modified by the concomitant infusion of 1, 2, or 4 mIU/min OT. These data show inhibition by elevated circulating OT levels of glucocorticoid-induced, but not basal, leptin secretion in normal weight subjects, suggesting a possible role for OT in the regulatory control of leptin. Furthermore, the results obtained in obese subjects indicate that this regulation is disrupted in obesity. PMID- 11061524 TI - A novel mutation causing complete thyroxine-binding globulin deficiency (TBG-CD Negev) among the Bedouins in southern Israel. AB - T4-binding globulin (TBG) is the major thyroid hormone transport protein in human serum. Inherited TBG abnormalities do not usually alter the metabolic status and are transmitted in X-linked inheritance. A high prevalence of complete TBG deficiency (TBG-CD) has been reported among the Bedouin population in the Negev (southern Israel). In this study we report a novel single mutation causing complete TBG deficiency due to a deletion of the last base of codon 38 (exon 1), which led to a frame shift resulting in a premature stop at codon 51 and a presumed truncated peptide of 50 residues. This new variant of TBG (TBG-CD-Negev) was found among all of the patients studied. We conclude that a single mutation may account for TBG deficiency among the Bedouins in the Negev. This report is the first to describe a mutation in a population with an unusually high prevalence of TBG-CD. PMID- 11061525 TI - High nocturnal melatonin in adolescents with chronic fatigue syndrome. AB - Decreased quality of sleep is frequently reported by chronic fatigue syndrome (CFS) patients. The pineal hormone melatonin is involved in regulation of sleep. We analyzed the nocturnal rise in melatonin in 13 adolescent CFS patients and 15 healthy age-matched controls. Saliva samples were collected at hourly intervals between 1700 and 0200 h. Nocturnal saliva melatonin levels were significantly higher in CFS patients, compared with controls, at midnight, 0100 h, and 0200 h (P < 0.001). No differences were observed in timing of melatonin increase in saliva between patients and controls. Time of sleep onset and duration of sleep did not differ significantly between patients and controls. However, all CFS patients and only one of the controls in our study group reported unrefreshing sleep. Our data demonstrate that sleep problems in adolescents with CFS are associated with increased melatonin levels during the first part of the night. Based on these data, we suggest that there is no indication for melatonin supplementation in adolescents with CFS. PMID- 11061526 TI - Retesting young adults with childhood-onset growth hormone (GH) deficiency with GH-releasing-hormone-plus-arginine test. AB - Within an appropriate clinical context, severe GH deficiency (GHD) in adults has to be defined biochemically by provocative testing of GH secretion. Patients with childhood-onset GHD need retesting in late adolescence or young adulthood to verify whether they have to continue recombinant human GH treatment. GHRH + arginine (GHRH+ARG) is the most reliable alternative to the insulin-induced hypoglycemia test (ITT) as a provocative test for the diagnosis of GHD in adulthood, provided that appropriate cut-off limits are assumed (normal limits, 16.5 microg/L as 3rd and 9.0 microg/L as 1st centile). We studied the GH response to a single GHRH (1 microg/kg iv) + ARG (0.5 g/kg iv) test in 62 young patients who had undergone GH replacement in childhood, based on the following diagnosis: 1) organic hypopituitarism with GHD (oGHD) In = 18: 15 male (M), 3 female (F); age, 26.8+/-2.2 yr; GH peak < 10 microg/L after two classical tests]; 2) idiopathic isolated GHD (iGHD) [n = 23 (15 M, 8 F); age, 23.0+/-1.5 yr; GH peak < 10 microg/L after two classical tests]; and 3) GH neurosecretory dysfunction (GHNSD) [n = 21 (10 M, 11 F); age, 25.1+/-1.6 yr; GH peak > 10 microg/L after classical test but mGHc < 3 microg/L]. The GH responses to GHRH+ARG in these groups were also compared with that recorded in a group of age-matched normal subjects (NS) [n = 48 (20 M, 28 F); age, 27.7+/-0.8 yr]. Insulin-like growth factor I levels in oGHD subjects (61.5+/-13.7 microg/L) were lower (P < 0.001) than those in iGHD subjects (117.2+/-13.1 microg/L); the latter were lower than those in GHNSD subjects (210.2+/-12.9 microg/L), which, in turn, were similar to those in NS (220.9+/-7.1 microg/L). The mean GH peak after GHRH+ARG in oGHD (2.8+/-0.8 microg/L) was lower (P < 0.001) than that in iGHD (18.6+/-4.7 microg/L), which, in turn, was clearly lower (P < 0.001) than that in GHNSD (31.3+/-1.6 microg/L). The GH response in GHNSD was lower than that in NS (65.9+/ 5.5 microg/L), but this difference did not attain statistical significance. With respect to the 3rd centile limit of GH response in young adults (i.e. 16.5 microg/L), retesting confirmed GHD in all oGHD, in 65.2% of iGHD, and in none of the GHNSD subjects. With respect to the 1st centile limit of GH response (i.e. 9.0 microg/L), retesting demonstrated severe GHD in 94% oGHD and in 52.1% of iGHD. All oGHD and iGHD with GH peak after GHRH+ARG lower than 9 microg/L had also GH peak lower than 3 microg/L after ITT. In the patients in whom GHD was confirmed by retesting, the mean GH peak after GHRH+ARG was higher than that after ITT (3.4+/-0.5 vs. 1.9+/-0.4). In conclusion, given appropriate cut-off limits, GHRH+ARG is as reliable as ITT for retesting patients who had undergone GH treatment in childhood. Among these patients, severe GHD in adulthood is generally confirmed in oGHD, is frequent in iGHD, but never occurs in GHNSD. PMID- 11061527 TI - Defect of villous cytotrophoblast differentiation into syncytiotrophoblast in Down's syndrome. AB - The syncytiotrophoblast (ST) is one of the major components of the human placenta, as it is involved in feto-maternal exchanges and the secretion of pregnancy-specific hormones. The aim of this study was to elucidate the formation and function of the ST in trisomy 21 (Down's syndrome). We first used the in vitro model of cytotrophoblast differentiation into ST. Cytotrophoblasts were isolated from 15 trisomy 21-affected placentas (12-35 weeks gestation) and 10 gestational age-matched control placentas. In vitro cytotrophoblasts isolated from normal placenta fused to form the ST. This was associated with an increase in transcript levels and in the secretion of hCG, human placental lactogen, placental GH, and leptin. In trisomy 21-affected placentas, we observed a defect (or a delay) in ST formation and a dramatic decrease in the synthesis and secretion of these hormones compared to those in cultured cells isolated from control age-matched placentas. These results were confirmed by a significant (P < 0.001) decrease in gene expression in total homogenates of trisomy 21-affected placentas compared to controls. These results will be of help in understanding the maternal hormonal markers of fetal trisomy 21 and the consequences of placental defects for fetal development. PMID- 11061528 TI - Two decades of screening for congenital hypothyroidism in The Netherlands: TPO gene mutations in total iodide organification defects (an update). AB - Presented is a cohort study to assess the nature and frequency of thyroid peroxidase (TPO) mutations in 45 patients (35 families) with congenital hypothyroidism due to a total iodide organification defect; incidence is 1:66,000 in The Netherlands. The presentation is consistently similar with a severe form of congenital hypothyroidism and also characterized by a complete and immediate release of accumulated radioiodide from the thyroid after sodium perchlorate administration. Sixteen different mutations were found, including eight novel mutations; the majority occurs in exons 8, 9, or 10. The GGCC insertion in exon 8 at nucleotide 1277, leading to an early termination signal in exon 9, is the most frequently occurring mutation. These mutations were detected in 29 families in both TPO alleles (13 homozygous and 16 compound heterozygous). In one family, partial maternal isodisomy of 2p was detected, in four families only one mutated TPO allele could be detected, and in one family no inactivating TPO mutation could be found. Because all patients clearly had the clinicopathologic features of a total iodide organification defect, we conclude that in these five families the mutations in the (other) alleles could be either located in the intronic sequences or in the promoter region. Mutations in the TPO gene result in total iodide organification defects. PMID- 11061529 TI - A comparison between the 1-microg adrenocorticotropin (ACTH) test, the short ACTH (250 microg) test, and the insulin tolerance test in the assessment of hypothalamo-pituitary-adrenal axis immediately after pituitary surgery. AB - The short ACTH stimulation test is an easy, reliable, and extensively used test in the assessment of the hypothalamo-pituitary-adrenal (HPA) axis. However, its use immediately after pituitary surgery is a matter of debate. The insulin tolerance test (ITT) is the gold standard in the evaluation of the HPA axis, but it is not always without side effects and may be unpleasant early after pituitary surgery. Our aim was to investigate the value of the 1-microg ACTH test in the assessment of the HPA axis early after pituitary surgery. We also aimed to determine the value of the 1-microg and 250-microg ACTH tests and the ITT in the estimation of HPA axis status after 3 months postoperatively. Nineteen patients subjected to pituitary tumor surgery were included in the study, and the ITT and the 1-microg and 250-microg ACTH tests were performed between the 4th and 11th days of surgery. The tests were repeated at the first month in 3 patients with subnormal peak cortisol responses (454, 125, and 301 nmol/L) and in 18 patients at the third month postoperatively. ACTH stimulation tests were performed by using 1 microg and 250 microg ACTH iv as a bolus injection, and blood samples were drawn at 0, 30, and 60 min for measurement of serum cortisol levels. The ITT was performed by using iv regular insulin, and serum glucose and cortisol levels were measured. The 1-microg and 250-microg ACTH stimulation tests and the ITT were performed consecutively. At least 48 h were allowed between each test. A peak serum cortisol level of 550 nmol/L or greater was considered as a normal response for both the ITT and the ACTH tests. The serum cortisol level was measured by RIA using commercial kits. Serum glucose was determined by glucose oxidase method. There were correlations between the peak cortisol response to the ITT and the 1-microg ACTH test (r = 0.39, P < 0.05) in the early postoperative period. No correlation was found between the ITT and the 250-microg ACTH test responses. In the early postoperative period, two patients showed normal cortisol responses (> or =550 nmol/L) to the 1-microg ACTH test and five patients showed normal cortisol responses to the 250-microg ACTH test among the six patients with subnormal cortisol responses to the ITT. Three patients with subnormal cortisol responses to ITT and baseline cortisol values less than 240 nmol/L showed normal HPA axis at the end of the first month. In the late postoperative period, at the third month, all the patients showed normal HPA axis. In the early postoperative period of pituitary surgery, the 1-microg ACTH test results are more concordant than the 250-microg ACTH test in comparison with the ITT. Our results also indicate that HPA axis dysfunction shown by ACTH stimulation tests and the ITT in early postoperative period may be normalized 1-3 months after surgery. For this reason, we think that dynamic tests including the ITT may not be useful early after pituitary surgery. PMID- 11061530 TI - Recombinant growth hormone (GH) therapy in GH-deficient adults: a long-term controlled study on daily versus thrice weekly injections. AB - Currently, replacement recombinant GH (rGH) therapy in GH-deficient (GHD) adults is performed in daily injections. This modality of treatment is not complied with by the totality of GHD patients, who are supposed to receive life-long replacement. The aim of our study was to compare daily vs. thrice weekly (TIW) rGH injection effects on lipid profile, body composition, bone metabolism, and bone density in 34 GHD patients (13 women and 21 men; median age, 39 yr; range, 30-55 yr) randomly assigned to different therapeutic regimens. Group A included 18 patients receiving daily rGH injections, and group B included 16 patients receiving TIW injections of rGH. The starting dose of rGH was 10 microg/kg x day in both groups. Subsequently, the dose was adjusted to maintain serum insulin like growth factor I (IGF-I) concentrations in the normal age-adjusted range. IGF I levels were assessed before and after 1, 3, 6, and 12 months of rGH treatment, and lipid profile, body composition, bone metabolism, and bone density were evaluated before and after 6 and 12 months of treatment. Thirty-four healthy subjects served as controls. In the basal condition, lipid profile, body composition, bone metabolism, and bone density were significantly different in patients compared to controls. Conversely, patients included in groups A and B had similar serum IGF-I levels, lipid profile, body composition, bone metabolism, and bone density. After 3 months of rGH treatment, IGF-I levels were normalized in 15 of 18 patients (83.3%) in group A and in 7 of 16 patients (43.7%) in group B (chi2 = 4.21; P = 0.04). At this time point, serum IGF-I levels in patients in group A (202+/-57.5 microg/L) were significantly higher than those in patients in group B (155+/-45.1 microg/L; P = 0.001). After 6 months of therapy, serum IGF-I levels were normalized in all patients and were similar in both groups (223+/ 35.2 vs. 212+/-41.4 microg/L, A vs. B, respectively). IGF-I levels remained normal until the 12-month follow-up. After 6 months of rGH replacement, total cholesterol, low density lipoprotein cholesterol, triglycerides, bioelectrical impedance, and body fat mass were significantly reduced, whereas high density lipoprotein cholesterol levels and lean body mass were significantly increased in both groups of patients, without any difference between them. No further change in lipid profile and body composition was observed after 12 months of treatment. Serum bone GLA protein and procollagen III levels were significantly increased after 6 months, and a downward trend was observed after 12 months of rGH replacement. However, a slight, but significant, increase in bone mineral density was observed in both groups only after 12 months (P = 0.0001). All patients in group B had good compliance to the TIW treatment, whereas 5 patients in group A had poor compliance to the treatment (chi2 = 3.2; P = 0.07). In conclusion, our randomized, prospective, and controlled study confirmed that rGH therapy with TIW injection regimen is effective in normalizing IGF-I levels and improving lipid profile, body composition, bone metabolism, and bone density. It also demonstrated that this efficacy is comparable to that observed in patients treated with daily rhGH therapy, with few side-effects and good compliance. PMID- 11061531 TI - A 1-year prospective study on the relationship between physical activity, markers of bone metabolism, and bone acquisition in peripubertal girls. AB - We conducted a 1-yr prospective study to evaluate the association between physical activity and biochemical markers of bone formation and resorption with bone mineral acquisition in 155 peripubertal Caucasian girls (51 gymnasts, 50 runners, and 54 nonathletic controls). The bone mineral density (BMD) of the femoral neck, the greater trochanter, and the lumbar spine were measured by dual energy x-ray absorptiometry. Serum biochemical markers of bone formation (osteocalcin, bone-specific alkaline phosphatase, amino-terminal propeptide of type I procollagen) and bone resorption (degradation product of C-terminal telopeptide of type I collagen) were measured. The 1-yr increase in BMD (adjusted for age, height, Tanner stage, BMD at baseline, and increases in height and weight) of the femoral neck was 0.037 g/cm2 x yr [95% confidence interval (CI), 0.019-0.051 g/cm2 x yr), and that of the greater trochanter was 0.020 g/cm2 x yr (95% CI, 0.003-0.039 g/cm2 x yr) greater in gymnasts than in controls. The corresponding figures for gymnasts compared with runners were 0.038 g/cm2 x yr (95% CI, 0.009-0.041 g/cm2 x yr) and 0.033 g/cm2 x yr (95% CI, 0.006 to 0.043 g/cm2 x yr). The figures for the lumbar spine did not differ significantly between study groups. The baseline serum concentrations of formation markers and resorption marker accounted for 2.3-12.8% (P < 0.05) of the variation in the 1-yr increase in BMD at the femoral neck and lumbar spine. However, there was no significant difference between the levels of adjusted baseline bone turnover markers of the gymnasts, runners, and controls. The present data add considerable support to the argument that high impact mechanical loading is extremely important and beneficial for the acquisition of BMD of the hip during peripubertal years. Our results indicate also that a high rate of bone turnover, reflected as elevated bone markers, is only weakly associated with the 1-yr bone gain in peripubertal girls. PMID- 11061532 TI - Tissue specificity of glucocorticoid sensitivity in healthy adults. AB - Contradicting data exist as to whether interindividual patterns in glucocorticoid (GC) sensitivity vary between different target tissues in humans. This study therefore measured GC sensitivity in 36 healthy subjects in three target tissues: the immune system; the cardiovascular system, and the hypothalamus-pituitary adrenal axis. For this purpose, dexamethasone inhibition of lipopolysaccharide induced interleukin-6 and tumor necrosis factor-alpha production in peripheral leukocytes, beclomethasone dipropionate-induced skin blanching, and suppression of cortisol levels after low-dose (0.5 mg) dexamethasone suppression test were determined in each subject. The results showed the expected glucocorticoid induced suppression of interleukin-6 and tumor necrosis factor-alpha production (both P < 0.001), dose-dependent skin blanching (P < 0.001), and suppression of salivary cortisol response to awakening (P < 0.001). However, neither simple correlations nor cluster analysis revealed a significant association among the three bioassays for GC sensitivity. In contrast to the idea that interindividual variation in GC sensitivity is an intrinsic trait affecting all tissues, these results suggest that this variability is target tissue specific in healthy subjects. PMID- 11061533 TI - Suppression of systemic, intramuscular, and subcutaneous adipose tissue lipolysis by insulin in humans. AB - In addition to sc and visceral fat deposits, muscle has been shown to contain relevant amounts of lipids whose breakdown is subject to hormonal regulation. The aim of the present study was to determine insulin dose-response characteristics of systemic, sc adipose tissue and muscle lipolysis in humans. We used a combination of isotopic (primed continuous infusion of [d5]glycerol) and microdialysis techniques (catheters placed in the anterior tibial muscle and sc abdominal adipose tissue) during a three-step hyperinsulinemic-euglycemic clamp (insulin infusion, 0.1, 0.25, 1.0 mU/kg x min) in 13 lean, healthy volunteers. The glycerol rate of appearance was used as the index for systemic lipolysis; interstitial glycerol concentrations were used as the index for muscle and sc adipose tissue lipolysis. The insulin concentrations resulting in a half-maximal suppression (EC50) of systemic lipolysis, adipose tissue, and muscle lipolysis were 51, 68, and 44 pmol/L, respectively (between one another, P < 0.001). For each compartment there were significant correlations between the EC50 and the insulin sensitivity index for glucose disposal (r > 0.67; P < 0.05). However, lipolysis (as percent of baseline) was similar during the first two insulin infusion steps, but was significantly lower in adipose (22+/-2%) than in muscle (53+/-4%; P < 0.001) during step 3. Although we have no direct measurement of interstitial insulin concentrations, we conclude that based on the EC50 values, muscle is more sensitive with respect to the net effect of circulating insulin (transendothelial transport plus intracellular action) on lipolysis than sc adipose tissue in terms of exerting its full suppression within the physiological insulin range. This could be important in muscle for switching from preferential utilization of free fatty acids to glucose in the postprandial state. Inadequate suppression of im lipolysis resulting in excessive local availability of free fatty acids may represent a novel mechanism contributing to the pathogenesis of impaired glucose disposal, i.e. insulin resistance, in muscle. PMID- 11061534 TI - Endocrine and metabolic responses in children with meningoccocal sepsis: striking differences between survivors and nonsurvivors. AB - To get insight in the endocrine and metabolic responses in children with meningococcal sepsis 26 children were studied the first 48 h after admission. On admission there was a significant difference in cortisol/ACTH levels between nonsurvivors (n = 8) and survivors (n = 18). Nonsurvivors showed an inadequate cortisol stress response in combination to very high ACTH levels, whereas survivors showed a normal stress response with significantly higher cortisol levels (0.62 vs. 0.89 micromol/L) in combination with moderately increased ACTH levels (1234 vs. 231 ng/L). Furthermore, there was a significant difference between nonsurvivors and survivors regarding pediatric risk of mortality score (31 vs. 17), TSH (0.97 vs. 0.29 mE/L), T3 (0.53 vs. 0.38 nmol/L), reverse T3 (rT3) (0.75 vs. 1.44 nmol/L), C-reactive protein (34 vs. 78 mg/L), nonesterified fatty acids (0.32 vs. 0.95 mmol/L), and lactate (7.3 vs. 3.2 mmol/L). In those who survived, the most important changes within 48 h were seen in a normalization of cortisol and ACTH levels, but without a circadian rhythm; a decrease of rT3 and an increase in the T3/rT3 ratio; and a decrease in the levels of the nonesterified free fatty acids and an unaltered high urinary nitrogen excretion. At this moment, it is yet unknown whether the hormonal abnormalities are determining factors in the outcome of acute meningococcal sepsis or merely represent secondary effects. Understanding the metabolic and endocrine alterations is required to design possible therapeutic approaches. The striking difference between nonsurvivors and survivors calls for reconsideration of corticosteroid treatment in children with meningococcal sepsis. PMID- 11061535 TI - Effect of two years of growth hormone and insulin-like growth factor-I suppression on prostate diseases in acromegalic patients. AB - The insulin-like growth factors (IGFs) have mitogenic effects on normal and tumoral prostate epithelial cells and have been suggested to be involved in prostate cancer. Moreover, chronic GH and IGF-I excess causes prostate overgrowth in patients with acromegaly. This study was designed to investigate whether the suppression of GH and IGF-I levels by surgery or pharmacotherapy could induce the regression of prostatic hyperplasia in acromegalic patients. To this end, prostate volume (PV) as well as the occurrence of prostatic diseases were studied by transrectal ultrasonography in 23 untreated acromegalic patients (with elevated GH and IGF levels). None of the patients reported symptoms due to prostatic disorders or obstruction. At study entry, prostate hyperplasia was found in half patients. After 2 yr, GH, IGF-I, and IGFBP-3 levels were decreased, whereas prostate-specific antigen levels did not change. PV was decreased in the 16 patients who were well controlled. Among the 6 patients with prostate hyperplasia at study entry who achieved disease control, 4 regained a normal PV at the end of the 2 yr of treatment, whereas none of the 5 patients with prostate hyperplasia at study entry and not achieving disease control normalized their PV. When patients were divided according to age, prostate volume decreased after 2 yr only in the 8 controlled patients aged below 50 yr, but not in those controlled and with age above 50 yr despite similar decrease in GH, IGF-I, and IGFBP3 levels. No clinical, transrectal ultrasonography, or cytological evidence of prostate cancer was detected during the study period. These data suggest that hyperplasia, but not cancer, is frequent in acromegalic men, and that the GH-IGF axis and age are independently associated with the development of this process. PMID- 11061536 TI - Effects of 7 years of growth hormone replacement therapy in hypopituitary adults. AB - Short-term studies of GH replacement in adult hypopituitarism have usually demonstrated beneficial effects on body composition and circulating lipids, with neutral or occasionally adverse effects on glucose tolerance. Fasting hyperinsulinemia has been reported. GH effects on cardiac function have been variable. The effects of long-term GH therapy, taking into account the consequences of increasing age, are not fully known. Thirty-three hypopituitary, initially middle-aged adults were studied over a 7-yr period; 12 patients took GH therapy (mean, 0.7 mg daily) continuously (group A); 11 took GH for only 6-18 months, a minimum of 5 yr previously (group B); and 10 patients never received GH therapy (group C). Other pituitary replacement was maintained. Effects on anthropometry, body composition (by bioimpedance analysis, total body potassium, and dual energy x-ray absorptiometry), circulating lipids, glucose and insulin concentrations, cardiac 2-dimensional and Doppler echocardiography, and exercise tolerance were assessed before and after the treatment period. Continuous GH therapy had no significant effect on body weight, but it prevented the increase in waist circumference and waist to hip ratio that occurred in the patients without GH substitution (waist to hip ratio, group A, 0.87+/-0.08 at baseline, 0.85+/-0.09 at 7 yr; group B, 0.89+/-0.11 at baseline, 0.94+/-0.11 at 7 yr; P < 0.005 for GH effect; group C, 0.87+/-0.10 at baseline, 0.92+/-0.10 at 7 yr; P < 0.005 for GH effect). A GH-induced decrease in subscapular skinfold thickness was also observed. By bioimpedance analysis, GH therapy caused an increase in total body water and fat-free mass, and a decrease in the percent body fat. Although changes occurred with time in all groups, no significant additional GH therapy effects were observed on glucose tolerance, insulin concentrations, lipid levels, cardiac dimensions, echocardiographic diastolic function, or exercise tolerance. In conclusion, prolonged GH substitution in middle-aged hypopituitary adults causes a sustained improvement in body composition. Other benefits, e.g. on lipid levels and exercise tolerance, were not apparent at 7 yr when comparisons were made with GH-untreated hypopituitary controls. Potentially adverse effects on glucose tolerance and insulinemia did not develop with prolonged GH therapy. PMID- 11061537 TI - Insulin resistance and substrate utilization in human endotoxemia. AB - Infection results in a state of insulin resistance, but the pathogenesis is poorly understood. Intravenous administration of bacterial lipopolysaccharide (LPS) has been used to mimic the febrile and systemic inflammatory responses to infection, but it is unclear whether LPS induces insulin resistance in man. To investigate the effects of LPS on insulin sensitivity and substrate utilization, we administered, in paired cross-over studies, either 20 U/kg Escherichia coli endotoxin or saline control to healthy volunteers (n = 6) 120 min after the start of a 10-h euglycemic hyperinsulinemic clamp (insulin infusion rate, 80 mU/m2 x min). LPS induced a fever, tachycardia, and mild arterial hypotension. Glucose utilization increased abruptly 120 min after LPS administration (+64.1+/-12.0%; P < 0.003), but then declined progressively, and insulin resistance was evident by 420 min (+1.9+/-3.5%; P < 0.05). The reduction in glucose utilization, like that observed in sepsis, was related to impaired nonoxidative glucose disposal and not abnormal glucose oxidation. The cortisol and GH responses to LPS were of sufficient duration and magnitude to explain the insulin resistance. LPS administration results in metabolic responses very similar to those observed in sepsis and could provide a useful model for the study of insulin resistance in human critical illness. PMID- 11061538 TI - Hormonal and metabolic effects of radiotherapy in acromegaly: long-term results in 128 patients followed in a single center. AB - Conventional radiotherapy is usually indicated in acromegaly when surgery fails to normalize GH secretion. However, the benefits of radiotherapy are delayed. This has raised questions about the potency of this treatment for reaching the safe GH level of 2.5 microg/L and for normalizing insulin-like growth factor I (IGF-I) levels, both of which are currently recommended as the therapeutic goal. To evaluate the long-term hormonal and metabolic effects of radiotherapy in acromegaly, a retrospective analysis was undertaken studying 128 patients followed for 11.5+/-8.5 yr (mean +/- SD) in a single center. The preradiation GH levels decreased as a function of time to 50% at 2 yr, 20% at 5 yr, and 10% at 10 yr. Basal GH levels below 2.5 microg/L were obtained in 7% of the patients at 2 yr, 35% at 5 yr, 53% at 10 yr, and 66% at 15 yr. A basal GH level below 2.5 microg/L was associated with suppression of GH below 2 microg/L during an oral glucose tolerance test and normalization of IGF-I levels in 9 of 10 patients. Preradiation GH levels was the sole factor that could predict the delay in GH fall to below 2.5 microg/L (P = 0.008). At the last follow-up, IGF-I levels were normalized in 79% of the patients (37 of 47; mean follow-up, 15.0+/-11.3 yr). In the 32 patients presenting with diabetes mellitus, improvement of glucose tolerance was associated with lower GH levels after treatment (35+/-78 microg/L in the group of 13 patients still presenting diabetes; 9+/-12 microg/L in the group of 4 patients with glucose intolerance; 5+/-8 microg/L in the 14 patients with normal glucose tolerance; P = 0.04). Ten years after termination of radiotherapy gonadotroph, thyreotroph and corticotroph deficiencies were observed in 80%, 78%, and 82% of the patients, respectively. In conclusion, conventional radiotherapy can reduce GH levels below the optimal level of 2.5 microg/L and normalize IGF-I levels in acromegaly. However, the incidence of late hypopituitarism is high, and the delay to obtain this safe GH secretory status can be long, depending on the preradiation GH level. These parameters should be considered when adjuvant therapy is needed after surgery. PMID- 11061539 TI - Body composition, blood pressure, and lipid metabolism before and during long term growth hormone (GH) treatment in children with short stature born small for gestational age either with or without GH deficiency. AB - To assess the effects of long-term continuous GH treatment on body composition, blood pressure (BP), and lipid metabolism in children with short stature born small for gestational age (SGA), body mass index (BMI), skinfold thickness measurements, systemic BP measurements, and levels of blood lipids were evaluated in 79 children with a baseline age of 3-11 yr with short stature (height SD score, < -1.88) born SGA (birth length SD-score, < -1.88). Twenty-two of the 79 children were GH deficient (GHD). All children participated in a randomized, double-blind, dose-response multicenter GH trial. Four- and 6-yr data were compared between two GH dosage groups (3 vs. 6 IU/m2 body surface/day). Untreated children with short stature born SGA are lean (mean BMI SD-score, -1.3; mean SD score skinfolds, -0.8), have a higher systolic BP (SD-score, 0.7) but normal diastolic BP (SD-score, -0.1), and normal lipids (total cholesterol, 4.7 mmol/L; low-density lipoprotein, 2.9 mmol/L; high-density lipoprotein, 1.3 mmol/L) compared with healthy peers. During long-term continuous GH treatment, the BMI normalized without overall changes in sc fat compared with age-matched references, whereas the BP SD-score and the atherogenic index decreased significantly. Although the mean 6-yr increase in height SD-score was significantly higher in the children receiving GH treatment with 6 IU/m2 x day (2.7) than in those receiving treatment with 3 IU/m2 day (2.2), no differences in the changes in BMI, skinfold measurements, BP, and lipids were found between the GH dosage groups. The pretreatment SD-scores for BMI, skinfold, and BP, as well as the lipid levels, were not significantly different between GHD and non-GHD children, but after 6 yr of GH treatment the skinfold SD-score and BP SD-score had decreased significantly more in the GHD than in the non-GHD children. Our data indicate that GH treatment has at least up to 6 yr positive instead of negative effects on body composition, BP, and lipid metabolism. In view of the reported higher risk of cardiovascular diseases in later life in children born SGA, further research into adulthood remains warranted. PMID- 11061540 TI - DR- and DQ-associated protection from type 1A diabetes: comparison of DRB1*1401 and DQA1*0102-DQB1*0602*. AB - The transmission disequilibrium test was used to analyze haplotypes for association and linkage to diabetes within families from the Human Biological Data Interchange type 1 diabetes repository (n = 1371 subjects) and from the Norwegian Type 1 Diabetes Simplex Families study (n = 2441 subjects). DQA1*0102 DQB1*0602 was transmitted to 2 of 313 (0.6%) affected offspring (P < 0.001, vs. the expected 50% transmission). Protection was associated with the DQ alleles rather than DRB1*1501 in linkage disequilibrium with DQA1*0102-DQB1*0602: rare DRB1*1501 haplotypes without DQA1*0102-DQB1*0602 were transmitted to 5 of 11 affected offspring, whereas DQA1*0102-DQB1*0602 was transmitted to 2 of 313 affected offspring (P < 0.0001). Rare DQA1*0102-DQB1*0602 haplotypes without DRB1*1501 were never transmitted to affected offspring (n = 6). The DQA1*0101 DQB1*0503 haplotype was transmitted to 2 of 42 (4.8%) affected offspring (P < 0.001, vs. 50% expected transmission). Although DRB1*1401 is in linkage disequilibrium with DQB1*0503, neither of the two affected children who carried DQA1*0101-DQB1*0503 had DRB1*1401. However, all 13 nonaffected children who inherited DQA1*0101-DQB1*0503 had DRB1*1401. In a case-control comparison of patients from the Barbara Davis Center, DQA1*0101-DQB1*0503 was found in 5 of 110 (4.5%) controls compared with 3 of 728 (0.4%) patients (P < 0.005). Of the three patients with DQB1*0503, only one had DRB1*1401. Our data suggest that both DR and DQ molecules (the DRB1*1401 and DQA1*0102-DQB1*0602 alleles) can provide protection from type 1A diabetes. PMID- 11061541 TI - Hashimoto's thyroiditis: countrywide screening of goitrous healthy young girls in postiodization phase in India. AB - Countrywide salt iodization, to prevent nutritional iodine deficiency, has been achieved in India recently. The current study was planned to evaluate the prevalence of goiter and thyroid autoimmunity and assess thyroid functional status in a cohort of 6283 healthy schoolgirls from different parts of the country in the postiodization phase. Goitrous girls (n = 1810; 28% of subjects) were investigated for serum T4 and TSH, antithyroid microsomal antibody (TMA) and antithyroglobulin antibody (TGA), urinary iodine excretion, and cytomorphology by fine-needle aspiration cytology (FNAC). FNAC carried out successfully in 764 goitrous girls revealed juvenile autoimmune thyroiditis (JAT) in 58 (7.5%), which included Hashimoto's thyroiditis in 43 (5.6%) and focal lymphocytic thyroiditis in 15 (1.9%). TMA and TGA estimated in 722 goitrous girls detected significantly positive titers of TMA (> or =1:1600) and TGA (> or =1:160) in 52 (7.2%) and 4 (0.55%) girls, respectively. Only 29 (67.4%) girls with Hashimoto's thyroiditis were TMA positive. In patients with FNAC-proven JAT, overt clinical and biochemical hypothyroidism was seen in three (6.5%) and subclinical hypothyroidism in seven (15%). Subclinical hyperthyroidism was detected in 5.1% cases of JAT, and none had overt hyperthyroidism. No definite correlation was seen between urinary iodine excretion and thyroid autoimmunity. PMID- 11061542 TI - Growth hormone secretagogue binding sites in peripheral human tissues. AB - The family of GH secretagogues (GHS) includes peptidyl (hexarelin) and nonpeptidyl (MK 0677) molecules possessing specific receptors in the brain, pituitary, and thyroid. GHS receptor subtypes have also been identified in the heart; and a gastric-derived peptide, named ghrelin, has recently been proposed as a natural ligand. Our aim was to investigate the presence of GHS receptors in a wide range of human tissues, by radioreceptor assay with [125I]Tyr-Ala hexarelin. GHS receptors were detected mainly in the myocardium, but they were also present (in order of decreasing binding activity) in adrenal, gonads, arteries, lung, liver, skeletal muscle, kidney, pituitary, thyroid, adipose tissue, veins, uterus, skin, and lymphnode. In contrast, negligible binding was found in parathyroid, pancreas, placenta, mammary gland, prostate, salivary gland, stomach, colon, and spleen. Hexarelin, MK 0677, and human ghrelin completely displaced the radioligand from binding sites of endocrine tissues, but MK 0677 and ghrelin were less potent than hexarelin. In nonendocrine tissues, both MK 0677 and ghrelin were inactive in displacement of [125I]Tyr-Ala hexarelin, whereas hexarelin was as active as a displacing agent in endocrine tissues. This study provides the first detailed analysis of the tissue localization of GHS receptors and suggests that a still unknown receptor subtype, specific for peptidyl GHS, may exist in the heart and in other tissues. PMID- 11061543 TI - Placental peroxisome proliferator-activated receptor-gamma is up-regulated by pregnancy serum. AB - Lipid metabolism plays an important role in normal pregnancy adaptation and in pathological pregnancy (e.g. preeclampsia). In the current studies we examined the expression of peroxisome proliferator-activated receptor-gamma (PPARgamma) in tissues and cells relevant to human pregnancy. We found that PPARgamma is expressed in placental cytotrophoblasts in vivo and in trophoblasts (primary and choriocarcinoma cells) and fetal endothelial cells in vitro. We characterized primary cytotrophoblasts and two cell lines with which to study PPARgamma regulation in human pregnancy. Like primary cytotrophoblasts, the choriocarcinoma cell line JEG-3 has endogenous PPARgamma expression. Normal positive and negative PPARgamma regulation was observed in the latter cells. We also created a new JEG 3-derived cell line (EP-JEG) by stable insertion of a PPAR response element luciferase reporter gene construct. Together, these cell lines are useful for studying PPARgamma expression and activation in human trophoblasts. We examined PPARgamma regulation in these cells by human serum and found that PPARgamma protein expression and activation are dramatically increased by sera from pregnant women. Preliminary characterization of the regulatory principle(s) is consistent with a prostanoid or fatty acid derivative. The results suggest that increased activation of PPARgamma may play an important role in maternal metabolism during human pregnancy. PMID- 11061544 TI - Luteinizing hormone-releasing hormone induces nuclear factor kappaB-activation and inhibits apoptosis in ovarian cancer cells. AB - More than 80% of human ovarian cancers express LHRH and its receptor as part of a negative autocrine mechanism of growth control. This study was conducted to investigate whether LHRH affects apoptosis in ovarian cancer. EFO-21 and EFO-27 ovarian cancer cells were treated with LHRH agonist Triptorelin or with cytotoxic agent Doxorubicin in the absence or presence of Triptorelin. Apoptotic cells were quantified by flow cytometry. Expression of nuclear factor kappa B (NFkappaB) was assessed by RT-PCR and immunoblotting. For determination of Triptorelin-induced NFkappaB activation, cells were transfected with a NFkappaB-secreted alkaline phosphatase reporter gene plasmid (pNFkappaB-SEAP) and cultured for 96 h with or without Triptorelin. The causal relation between Triptorelin-induced NFkappaB activation and Triptorelin-induced protection against apoptosis was investigated using SN50, an inhibitor for nuclear translocation of activated NFkappaB. Apoptosis induction by Triptorelin was never observed. Treatment with Doxorubicin (1 nmol/L) for 72 h increased the percentage of apoptotic cells in EFO-21 and EFO 27 ovarian cancer cell lines to 31% or 34%, respectively. In cultures treated simultaneously with Triptorelin (100 nmol/L), the percentage of apoptotic cells was reduced significantly, to 17% or 18%, respectively (P < 0.001). RT-PCR and immunoblotting experiments showed that NFkappaB subunits p50 and p65 were expressed by ovarian cancer cell lines EFO-21 and EFO-27. When EFO-21 or EFO-27 cells were transfected with pNFkappaB-SEAP and subsequently treated with Triptorelin (100 nmol/L), NFkappaB-induced SEAP expression increased 5.3-fold or 4.7-fold, respectively (P < 0.001). Triptorelin-induced reduction of Doxorubicin induced apoptosis was blocked by SN50-mediated inhibition of NFkappaB translocation into the nucleus. We conclude that LHRH induces activation of NFkappaB and thus reduces Doxorubicin-induced apoptosis in human ovarian cancer cells. This possibility to protect ovarian cancer cells from programmed cell death is an important feature in LHRH signaling in ovarian tumors, apart from the inhibitory interference with the mitogenic pathway. PMID- 11061545 TI - Regulation of insulin-like growth factor-binding protein 1 by hypoxia and 3',5' cyclic adenosine monophosphate is additive in HepG2 cells. AB - Insulin-like growth factor-binding protein 1 (IGFBP-1) is important in regulating minute-to-minute IGF bioavailability in the circulation and is primarily an inhibitor of IGF action systemically and in most cellular systems. Understanding regulation of IGFBP-1 is, thus, important in understanding regulation of IGF actions. The IGFBP-1 promoter contains a cAMP response element, and cAMP stimulates IGFBP-1 gene expression at the transcriptional level. Recently, we have found three consensus sequences for the hypoxia response element in intron 1 of the IGFBP-1 gene. Herein, we have investigated the effects of hypoxia and a cAMP analog, 8-bromoadenosine-3',5'-cyclic monophosphate (8-Br-cAMP), on IGFBP-1 expression in HepG2 cells, a model system for IGFBP-1 gene regulation. HepG2 cells were exposed to normoxia (20% pO2) or hypoxia (2% pO2) for 24 h in the absence or presence of 8-Br-cAMP (0.1, 0.5, and 1 mM). Western ligand blotting revealed IGFBP-1 as the predominant IGFBP in HepG2-conditioned media, which increased in a dose-dependent manner after incubation with 8-Br-cAMP in normoxia and hypoxia (3-fold and 7-fold at 1 mM, respectively). Under hypoxic, compared with normoxic, conditions, IGFBP-1 protein and messenger RNA (mRNA) levels increased approximately 10-fold and 20-fold, respectively. In normoxia, 8-Br-cAMP stimulated IGFBP-1 protein and mRNA levels in a dose-dependent manner (7-fold and 10-fold at 1 mM). Hypoxia and 8-Br-cAMP showed additive stimulatory effects on IGFBP-1 protein and mRNA levels (35-fold and 50-fold at 1 mM) that were time and dose dependent. Primary transcripts of IGFBP-1 mRNA were increased concordantly with IGFBP-1 mRNA. The half-life of the IGFBP-1 mRNA was markedly increased (approximately 6-fold) by hypoxia, and cAMP minimally enhanced this effect. These results demonstrate that hypoxia and compounds that increase intracellular cAMP additively regulate IGFBP-1 gene expression by transcriptional and posttranscriptional mechanisms. Regulation of IGFBP-1 mRNA and protein by cAMP and hypoxia may be important for understanding the physiologic and pathophysiologic roles of IGFBP-1. PMID- 11061546 TI - Hormonal regulation of estrogen receptor alpha and beta gene expression in human granulosa-luteal cells in vitro. AB - Estrogen is one of the major sex steroid hormones that is produced from the human ovary, and its actions are established to be a receptor-mediated process. Despite the demonstration of estrogen receptor (ER) expression, little is known regarding the regulation of ER in the human ovary. In the present study we investigated the expression and hormonal regulation of ERalpha and ERbeta in human granulosa luteal cells (hGLCs). Using RT-PCR amplification, both ERalpha and ERbeta messenger ribonucleic acid (mRNA) were detected from hGLCs. Northern blot analysis revealed that ERalpha is expressed at a relatively lower level than ERbeta. Basal expression studies indicated that ERalpha mRNA levels remain unchanged, whereas ERbeta mRNA levels increased with time in culture in vitro, suggesting that ERbeta is likely to play a dynamic role in mediating estrogen action in hGLCs. The regulation of ERalpha and ERbeta expression by hCG was examined. hCG treatment (10 IU/mL) significantly attenuated the ERalpha (45%; P < 0.01) and ERbeta (40%; P < 0.01) mRNA levels. The hCG-induced decrease in ERalpha and ERbeta expression was mimicked by 8-bromo-cAMP (1 mmol/L) and forskolin (10 micromol/L) treatment. Additional studies using a specific protein kinase A (PKA) inhibitor (adenosine 3',5'-cyclic monophosphorothioate, Rp-isomer, triethylammonium salt) and an adenylate cyclase inhibitor (SQ 22536) further implicated the involvement of the cAMP/PKA signaling pathway in hCG action in these cells. The hCG-induced decrease in ERalpha and ERbeta mRNA levels was prevented in the presence of these inhibitors. Next, the effect of GnRH on ER expression was studied. Sixty-eight percent (P < 0.001) and 60% (P < 0.001) decreases in ERalpha and ERbeta mRNA levels, respectively, were observed after treatment with 0.1 micromol/L GnRH agonist (GnRHa). Pretreatment of the cells with a protein kinase C (PKC) inhibitor (GF109203X) completely reversed the GnRHa induced down-regulation of ERalpha and ERbeta expression, suggesting the involvement of PKC in GnRH signal transduction in hGLCs. In agreement with the semiquantitative RT-PCR results, Western blot analysis detected a decrease in ERalpha and ERbeta proteins levels in hGLCs after treatment with hCG (10 IU/mL), GnRH (0.1 micromol/L), 8-bromo-cAMP (1 mmol/L), forskolin (10 micromol/L), or phorbol 12-myristate 13 acetate (10 micromol/L). Functionally, we demonstrated an inhibition of progesterone production in hGLCs in vitro by 17beta-estradiol, and this inhibitory effect was eliminated by pretreatment of 10 IU/mL hCG or 0.1 micromol/L GnRHa for 24 h before 17beta-estradiol administration. In summary, we observed a differential expression of ERalpha and ERbeta mRNA in hGLCs in vitro. The demonstration of hCG- and GnRHa-induced down-regulation of ERalpha and ERbeta gene expression suggests that hCG and GnRH may contribute to the control of granulosa-luteal cell function. Furthermore, our data suggest that the effects of hCG and GnRH on ERalpha and ERbeta expression in hGLCs are mediated in part by activation of PKA and PKC signaling pathways, respectively. PMID- 11061547 TI - Estrogen receptor beta (ERbeta) messenger ribonucleic acid (mRNA) expression within the human forebrain: distinct distribution pattern to ERalpha mRNA. AB - Estrogen has been shown to influence several brain functions as well as the expression of neuropsychiatric diseases. To date, two estrogen receptor (ER) subtypes have been identified, ERalpha and ERbeta. ERalpha messenger ribonucleic acid (mRNA) distribution in the human forebrain was recently characterized, and the highest expression was found in restricted areas of the amygdala and hypothalamus. However, no information exists with regard to ERbeta mRNA distribution in the human brain. To this end, the anatomical distribution pattern of ERbeta mRNA expression in the human forebrain was investigated in the present study. Overall, the ERbeta mRNA hybridization signal was relatively low, but the most abundant ERbeta mRNA areas were the hippocampal formation (primarily the subiculum), claustrum, and cerebral cortex; expression was also present in the subthalamic nucleus and thalamus (ventral lateral nucleus). In contrast to ERalpha (studied on adjacent brain sections), ERbeta mRNA expression was low in the hypothalamus and amygdala. Based on the revealed anatomical distribution of the human ERbeta gene expression, a putative role for ERbeta in the modulation of cognition, memory, and motor functions is suggested. PMID- 11061548 TI - Insights from a successful case of intrahepatic islet transplantation into a type 1 diabetic patient. AB - We report a case of long-term (>4 yr) successful intrahepatic islet transplantation into a type 1 diabetic patient chronically immunosuppressed for a prior kidney graft. The exogenous insulin requirement decreased progressively after transplantation, and insulin treatment was withdrawn at 6 months. Glycosylated hemoglobin levels were in the normal range at 1 and 2 yr (5.3%) and increased slightly above the upper normal limit at 3 and 4 yr (6.3% and 6.4%). Fasting C peptide levels remained stable during the entire follow-up, but the proinsulin to insulin ratios increased dramatically at yr 3. Glycemic levels after an oral glucose tolerance test showed a diabetic profile at 1 yr, a normal profile at 2 yr, and an impaired glucose tolerance profile at 3 yr. Intravenous glucose tolerance test-induced first phase insulin release, present at 1 and 2 yr, disappeared at 3 yr. Diabetes-related autoantibodies (islet cell antibodies, glutamic acid decarboxylase antibodies, and tyrosine phosphatase-like protein antibodies) were undetectable before transplantation and remained so during the entire follow-up. The patient died of myocardial infarction 50 months after transplantation while she was still in good metabolic control (glycosylated hemoglobin, <6.8%) in the absence of exogenous insulin administration. The autoptic liver showed well granulated islets, richly vascularized and without evidence of lympho-mononuclear cell infiltration. The morphometrically extrapolated intrahepatic beta-cell mass was 99.9 mg. In conclusion, this successful islet graft showed a bell-shaped clinical effect, maximal at 2 yr after transplantation, followed by a slow progressive decline. The absence of allo- and autoreactivities against the transplanted islets points to a nonimmune mediated beta-cell loss as the cause of graft functional deterioration. PMID- 11061549 TI - Effects of norplant on endometrial tissue factor expression and blood vessel structure. AB - Abnormal uterine bleeding after Norplant administration is primarily responsible for the high discontinuation rate of this safe and effective long-acting implantable progestin-only contraceptive agent. Although tissue factor (TF) is the primary initiator of hemostasis, previous studies indicated that Norplant associated bleeding persists despite relatively high TF levels in the stromal compartment. Recently, we determined that progestin-enhanced TF expression during decidualization of human endometrial stromal cells involves both the epidermal growth factor receptor and progesterone receptor (PR]. The current study evaluated TF levels in endometrial bleeding (BL) and nonbleeding (NBL) sites obtained by camera-guided hysteroscopy during Norplant contraception. After 1 yr of therapy, immunohistochemical TF levels were unexpectedly higher at BL than at NBL sites. Use of immunohistochemistry and Western blotting indicated that both sites displayed elevated epidermal growth factor receptor levels and that the BL sites exhibited high levels of the PR, as well as the PR(A) and the PR(B) isoforms. Microscopic examination of 1-yr biopsies revealed that significantly larger numbers of enlarged, distended vessels were present in BL, compared with NBL sites. Elevated TF levels and abnormally enlarged blood vessels in the BL sites are consistent with the recently discovered angiogenic role of TF. By promoting aberrant angiogenesis, chronic endometrial overexpression of TF could produce fragile vessels, which are at increased risk to bleed. Analysis of endometrial BL and NBL sites, during Norplant contraception, offers the potential of elucidating local mechanisms that control enhanced TF expression, leading to abnormal angiogenesis at specific endometrial sites. PMID- 11061550 TI - Genetic and histologic studies of somatomammotropic pituitary tumors in patients with the "complex of spotty skin pigmentation, myxomas, endocrine overactivity and schwannomas" (Carney complex). AB - Carney complex (CNC) is a familial multiple neoplasia and lentiginosis syndrome with features overlapping those of McCune-Albright syndrome (MAS) and other multiple endocrine neoplasia (MEN) syndromes, MEN type 1 (MEN 1), in particular. GH-producing pituitary tumors have been described in individual reports and in at least two large CNC patient series. It has been suggested that the evolution of acromegaly in CNC resembles that of the other endocrine manifestations of CNC in its chronic, often indolent, progressive nature. However, histologic and molecular evidence has not been presented in support of this hypothesis. In this investigation, the pituitary glands of eight patients with CNC and acromegaly [age, 22.9+/-11.6 yr (mean +/- SD)] were studied histologically. Tumor DNA was used for comparative genomic hybridization (CGH) (four tumors). All tumors stained for both GH and prolactin PRL (eight of eight), and some for other hormones, including alpha-subunit. Evidence for somatomammotroph hyperplasia was present in five of the eight patients in proximity to adenoma tissue; in the remaining three only adenoma tissue was available for study. CGH showed multiple changes involving losses of chromosomal regions 6q, 7q, 11p, and 11q, and gains of 1pter-p32, 2q35-qter, 9q33-qter, 12q24-qter, 16, 17, 19p, 20p, 20q, 22p and 22q in the most aggressive tumor, an invasive macroadenoma; no chromosomal changes were seen in the microadenomas diagnosed prospectively (3 tumors). We conclude that, in at least some patients with CNC, the pituitary gland is characterized by somatotroph hyperplasia, which precedes GH-producing tumor formation, in a pathway similar to that suggested for MAS-related pituitary tumors. Three GH-producing microadenomas showed no genetic changes by CGH, whereas a macroadenoma in a patient, whose advanced acromegaly was not cured by surgery, showed extensive CGH changes. We speculate that these changes represent secondary and tertiary genetic "hits" involved in pituitary oncogenesis. The data (1) underline the need for early investigation for acromegaly in patients with CNC; (2) provide a molecular hypothesis for its clinical progression; and (3) suggest a model for MAS- and, perhaps, MEN 1-related GH-producing tumor formation. PMID- 11061551 TI - Quantitative determination of sst2 gene expression in neuroblastoma tumor predicts patient outcome. AB - Neuroblastoma (NB) is the most common pediatric neuroendocrine tumor, and it is characterized by a quite variable clinical course. We previously found a great variability in the expression of somatostatin receptor type 2 (sst2) in several human NB cell lines and primary tumors. In this report we investigated whether expression of sst2 is somehow related to clinical outcome. We performed a retrospective study on 54 patients with a maximum follow-up of 100 months. The concentration of specific messenger ribonucleic acid (mRNA) for sst2 was measured by competitive RT-PCR and validated, in a small subset of samples, by quantitative imaging of gene (in situ hybridization) and protein (immunohistochemistry) expression. We found that sst2 mRNA was variably expressed in all NB tumors (range, 2.5 x 10(5) to 8 x 10(9) molecules/microg RNA) with a relevant reduction in the more advanced stage (P < 0.01). Analysis of Kaplan Meier curves indicated that sst2 expression is positively related to the overall (P < 0.0001) and event-free (P < 0.0001) survival. Expression of sst2 was negatively related to tumor stage (P < 0.02) and MYCN amplification (P < 0.001), a poor prognostic factor. However, the prognostic information derived from sst2 is apparently independent from MYCN amplification, as assessed by stratifying sst2 values according to MYCN. In addition, the expression of sst2 was the only significant prognostic factor (P < 0.02) when it was included in a multivariate model containing other well known prognostic factors such as age, stage, and MYCN amplification. Hence, we propose that sst2 expression represents a new prognostic marker for NB. The main clinical value of a quantitative measure of sst2 lies in its ability to detect patients at low risk, independently from other prognostic factor, including MYCN amplification. PMID- 11061552 TI - Peroxisome proliferator-activated receptor-gamma modulates differentiation of human trophoblast in a ligand-specific manner. AB - The ligand-dependent nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) regulates the differentiation of several tissues and cell types. PPARgamma was recently determined to be essential for murine placental development and differentiation. We therefore assessed the influence of PPARgamma on differentiation of human placental trophoblasts. We initially used immunohistochemistry to examine term human placentas for PPARgamma expression and found that PPARgamma is present in syncytiotrophoblasts and cytotrophoblasts in placental villi. We correlated the expression of PPARgamma with differentiation of primary human trophoblasts and found that 8-bromo-cAMP, a known enhancer of trophoblast differentiation, stimulates PPARgamma activity, but has no effect on PPARgamma expression. We demonstrated that the PPARgamma ligand 15-deoxy delta12,14-prostaglandin J2 (15deltaPGJ2) and the thiazolidinedione troglitazone stimulate PPARgamma activity in the trophoblast cell line BeWo. Importantly, whereas exposure of cultured primary trophoblasts to troglitazone enhances biochemical and morphological trophoblast differentiation, 15deltaPGJ2 diminishes trophoblast differentiation. Furthermore, 15deltaPGJ2, but not troglitazone, up regulates p53 expression and promotes trophoblast apoptosis. These data indicate that PPARgamma is expressed in human placental trophoblasts, and that ligand specific activation of PPARgamma results in opposing effects on trophoblast differentiation. Our results suggest that PPARgamma plays an important role in placental differentiation during human pregnancy. PMID- 11061553 TI - Subcellular distribution of somatostatin sst2A receptors in human tumors of the nervous and neuroendocrine systems: membranous versus intracellular location. AB - The distribution of the sst2A receptor was investigated, using immunohistochemistry, with the specific antipeptide antibody R2-88, in a total of 120 tumors of the nervous and the neuroendocrine systems, including small-cell lung carcinomas, medulloblastomas, neuroblastomas, pheochromocytomas, and paragangliomas. The great majority of the tumor samples, frozen or formalin fixed, showed a positive immunohistochemical staining with R2-88, and an excellent correlation with receptor autoradiography using 125I[Tyr3]-octreotide. Whereas small-cell lung carcinomas and medulloblastomas had a predominantly plasma membrane staining, pheochromocytomas and neuroblastomas had variable ratios of cell surface and intracellular staining. Strikingly, a preferentially cytoplasmic staining was seen in tumors with a high level of somatostatin gene expression, whereas a more plasma membranous staining was seen in tumors lacking somatostatin messenger RNA. Specificity of both the plasma membrane and the cytoplasmic staining pattern was confirmed in immunoblots, which showed the immunoreactive receptor migrating as a characteristic 70-kDa broad band. In both immunohistochemical and immunoblotting experiments, staining was abolished by antibody blockade with 100 nM antigen peptide. These results describe, for the first time, the localization of the sst2A receptor protein in human small-cell lung carcinomas, medulloblastomas, neuroblastomas, and paragangliomas. Moreover, it is the first report investigating possible causes for distinct subcellular localizations of sst2A in human tissues. We show that the subcellular distribution of the receptor may be dependent on the surrounding somatostatin concentration, consistent with both the known effect of somatostatin to cause sst2A receptor internalization and an autocrine regulation of tumors by the peptide they produce. Moreover, our demonstration that the sst2A receptor can be identified in this group of tumors using simple immunohistochemical methods in formalin-fixed, paraffin-embedded material opens numerous diagnostic, therapeutic, and prognostic opportunities. PMID- 11061554 TI - A role for activin A and betacellulin in human fetal pancreatic cell differentiation and growth. AB - Activin A (Act.A), a member of the transforming growth factor beta family of secreted proteins, has been implicated in the regulation of growth and differentiation of various cell types. Betacellulin (BTC), a member of the epidermal growth factor family, converts exocrine AR42J cells to insulin expressing cells when combined with Act.A. We have used primary cultures of human fetal pancreatic tissue to identify the effects of Act.A and/or BTC on islet development and growth. Exposure to Act.A resulted in a 1.5-fold increase in insulin content (P < 0.005) and a 2-fold increase in the number of cells immunopositive for insulin (P < 0.005). The formation of islet-like cell clusters, containing mainly epithelial cells, during a 5-day culture, was stimulated 1.4-fold by BTC (P < 0.05). BTC alone caused a 2.6-fold increase in DNA synthesis (P < 0.005). These data suggest that Act.A induces endocrine differentiation, whereas BTC has a mitogenic effect on human undifferentiated pancreatic epithelial cells. PMID- 11061555 TI - Tyrosines 1015 and 1062 are in vivo autophosphorylation sites in ret and ret derived oncoproteins. AB - Point mutations of the RET receptor tyrosine kinase are responsible for the inheritance of multiple endocrine neoplasia (MEN) type 2 syndromes and are also present in a fraction of sporadic medullary thyroid carcinomas. Somatic rearrangements of the RET gene generating the chimeric RET/papillary thyroid carcinoma (PTC) oncogenes are the predominant molecular lesions associated with papillary carcinoma, the most frequent thyroid malignancy in humans. Oncogenic mutations cause constitutive activation of the kinase function of RET, which, in turn, results in the autophosphorylation of RET tyrosine residues critical for signaling. In vitro kinase assays previously revealed six putative RET autophosphorylation sites. The aim of the present study was to assess the phosphorylation of two such residues, tyrosines 1015 and 1062 (Y1015 and Y1062), in the in vivo signaling of RET and RET-derived oncogenes. Using phosphorylated RET-specific antibodies, we demonstrate that both Y1015 and Y1062 are rapidly phosphorylated upon ligand triggering of RET. Moreover, regardless of the nature of the underlying activating mutation, the concomitant phosphorylation of Y1015 and Y1062 is a common feature of the various oncogenic RET products (MEN2A, MEN2B, and PTC). This study shows that Ab-pY1062 is a useful tool with which to detect activated RET in human tumor cells and surgical samples. Finally, the microinjection of Ab-pY1062 antibodies into living cells demonstrates that Ret/PTC1 signaling is required to maintain the mitogenesis of a human carcinoma cell line expressing the Ret/PTC1 oncoprotein. PMID- 11061556 TI - Early identification of children predisposed to low peak bone mass and osteoporosis later in life. AB - The amount of bone that is gained during adolescence is the main contributor to peak bone mass, which, in turn, is a major determinant of osteoporosis and fracture risk in the elderly. We examined whether computed tomography measurements for the density and the volume of bone in the axial and the appendicular skeletons could be tracked through puberty in 40 healthy white children (20 girls and 20 boys). Longitudinal measurements of the cross-sectional area and cancellous bone density of the vertebral bodies and the cross-sectional and cortical bone areas of the femurs at the beginning of puberty accounted for 62-92% of the variations seen at sexual maturity; on average, 3 yr later. When baseline values for these bone traits were divided into quartiles, a linear relation across Tanner stages of sexual development was observed for each quartile in both girls and boys. The regression lines differed among quartiles for each trait, paralleled each other, and did not overlap. Thus, we are now in a position to identify those children who are genetically prone to develop low values for peak bone mass and toward whom osteoporosis prevention trials should be geared. PMID- 11061557 TI - Expression of vascular endothelial growth factor and its receptors in the human corpus luteum during the menstrual cycle and in early pregnancy. AB - To investigate the possible role of vascular endothelial growth factor (VEGF) and its receptors in the human corpus luteum (CL), expression of VEGF and its receptors, the fms-like tyrosine kinase and the kinase insert domain-containing region (KDR), was analyzed in the CL during the menstrual cycle and in early pregnancy. Immunohistochemistry revealed that VEGF was localized in luteal cells and both flt-1 and KDR were also localized in luteal cells, in addition to vascular endothelial cells. Messenger RNA (mRNA) expression of VEGF, flt-1, and KDR remained constant in the CL during the luteal phase and was lower in the regression phase. In the pregnant CL, VEGF mRNA expression was higher compared with that in the midluteal phase, and mRNA expression of both flt-1 and KDR was the same as that in the midluteal phase. Western blot analyses revealed that the change in protein expression of VEGF, flt-1, and KDR was similar to that in their mRNA expression. To study the effect of human CG (hCG) on VEGF expression in the CL, corpora lutea obtained from the midluteal phase were incubated with hCG (1 IU/ml) for 6 h. hCG increased the expression of mRNA and protein of VEGF. In conclusion, VEGF and its receptors may play important roles in development and function of the CL, and VEGF may exert a paracrine-autocrine role in regulating luteal function. hCG may act to prolong the life span of the CL by stimulating VEGF expression when pregnancy occurs. PMID- 11061558 TI - Expression of transcription factor GATA-4 during human testicular development and disease. AB - GATA-4 is a highly conserved transcription factor that plays a critical role in regulating embryonic morphogenesis and cellular differentiation. Given the emerging role of GATA-4 in the development and function of murine gonads, we have now studied its role in human testis. We find that GATA-4 is expressed from early human fetal testicular development to adulthood. This transcription factor is evident in Sertoli cells through fetal and postnatal development. Expression of GATA-4 in Sertoli cells peaks at 19-22 weeks gestation at the time of high circulating fetal FSH. In Leydig cells, GATA-4 is expressed during fetal period and after puberty, coinciding with the periods of active androgen synthesis in the testis; this suggests a link between GATA-4 and steroidogenesis. Also, fetal germ cells and prepubertal spermatogonia express GATA-4, and it is down-regulated in these cells after puberty. As hormonal regulation of GATA-4 in human testis was not possible to study directly, we used testicular samples from patients who had endocrine abnormalities or were hormonally treated. Testicular expression of GATA-4 in hCG-treated cryptorchidism does not differ from that in controls. In androgen resistance, GATA-4 expression in Sertoli and germ cells is weak or totally absent. GATA-4 protein is abundantly present in Sertoli and Leydig cell tumors, suggesting a relationship to tumorigenesis or tumor progression in somatic cell-derived testicular neoplasms. PMID- 11061559 TI - Leukemia inhibitory factor (LIF) stimulates the human HLA-G promoter in JEG3 choriocarcinoma cells. AB - HLA-G is a non-classic class I MHC molecule specifically expressed by human invasive cytotrophoblast cells, which has been suggested to play a role in facilitating the immune tolerance of the conceptus. So far, very little is known about the regulation of the human HLA-G gene. The present study was, thus, designed to investigate the regulation of the human HLA-G promoter. JEG3 choriocarcinoma cells, which express HLA-G endogenously, were used as a model. A 890 bp fragment of the human HLA-G promoter was amplified by nested PCR from genomic DNA, cloned into pCR-Script and, after sequencing, subcloned into pGL3 Luc in front of the luciferase reporter gene. This vector was then used in transient transfection experiments in JEG3 cells. Parallel transfection experiments were performed using an alpha subunit (alphaSU)-Luc reporter plasmid as a control. Using this system, several potential modulating substances were tested in different concentrations and for different periods of time: phorbol ester (TPA), cAMP, IFNgamma, IL-1, and leukemia inhibitory factor (LIF), with only LIF administration resulting in induction of the HLA-G promoter. LIF treatment also resulted in induction of HLA-G mRNA. JEG3 cells are shown to possess LIF receptors. LIF is a pleiotropic cytokine produced at the maternal fetal interface which has been shown to play an essential role in implantation in mice. LIF is produced in high amounts by the human endometrium and the trophoblast itself, and LIF receptors are present on cytotrophoblast cells. LIF could, thus, play a role in modulating HLA-G production and immune tolerance at the maternal-fetal interface. PMID- 11061560 TI - Rarity of anti- Na+/I- symporter (NIS) antibody with iodide uptake inhibiting activity in autoimmune thyroid diseases (AITD). AB - The search for antibody against the Na+/I- symporter (NIS) has seen conflicting results over the years. Prior to cloning of NIS, Raspe et al found iodide uptake inhibiting sera were rare in autoimmune thyroid diseases (AITD) while post cloning, others reported the presence of antibody in 12-15% of Hashimoto's thyroiditis (HT) and 30-84% of Graves' disease (GD). To evaluate the role of NIS as a potential antigen in AITD, a stable COS 7 cell line expressing high level of functional hNIS was established which allowed the screening of large number of sera for iodide uptake inhibiting activity in a 96-well plate format. Five hundred and fourteen serum samples taken from normal subjects and patients with AITD, non-autoimmune thyroid diseases, and non-thyroid autoimmune diseases were assayed for presence of iodide uptake inhibiting activity. Under the influence of these sera, iodide uptake showed a normal frequency distribution and diminution of uptake 2 SDs below the mean of controls was observed with 14 sera. Among these, 7 that were available for further study were re-evaluated after dialysis and/or Ig G extraction. All 7 sera lost their iodide uptake inhibiting activity, indicating that the effects were not antibody mediated and unknown serum factors had been responsible. In conclusion, contrary to previous results, the present study indicates that antibodies capable of modulating NIS activity are rare in AITD. PMID- 11061561 TI - Expression of pro-EPIL peptides encoded by the insulin-like 4 (INSL4) gene in chromosomally abnormal pregnancies. AB - The recent development of a specific immunoassay based on monoclonal antibodies directed to chain C and chain A of early placenta insulin-like peptide (EPIL) encoded by the INSL4 gene, has made it possible to demonstrate pro-EPIL peptide expression during normal pregnancy. In the present study, we report on the expression of pro-EPIL peptides in chromosomally abnormal pregnancies, namely trisomy 21 and 18. EPIL peptide levels were measured in amniotic fluid (AF) and maternal serum (MS) from pregnancies with trisomy 21 (n=16) or 18 (n=14) and compared to levels detected in AF and MS from 33 chromosomally normal pregnancies between 12 and 32 weeks of gestation. Pro-EPIL peptide levels were significantly higher in amniotic fluids from T21 than in AF from chromosomally normal pregnancies (mean pro-EPIL levels +/- SEM, 449+/-129.2 ng/mL vs 137+/-29.6 ng/mL, P = 0.0195), whereas there was only a trend towards an increase in pro-EPIL peptide levels in maternal serum. In a limited matched gestational age range (15 to 17 weeks), it was confirmed that pro-EPIL peptide levels were significantly higher in AF from T21 pregnancies (644.0+/-155.9 ng/mL, n = 11) than in AF from normal pregnancies (177.8+/-39.0 ng/mL, n = 12; P < 0.0001). Interestingly, the expression patterns of pro-EPIL peptides, human chorionic gonadotropin (hCG) and its free subunits were parallel in T21 pregnancies as recently observed in normal pregnancies. These results are in line with previous observations suggesting that the biosynthesis of both hCG and EPIL follows common regulation pathways. PMID- 11061562 TI - How the assembly dynamics of the nematode major sperm protein generate amoeboid cell motility. AB - Nematode sperm are amoeboid cells that use a major sperm protein (MSP) cytoskeleton in place of a conventional actin cytoskeleton to power their amoeboid motility. In these simple, specialized cells cytoskeletal dynamics is tightly coupled to locomotion. Studies have capitalized on this feature to explore the key structural properties of MSP and to reconstitute motility both in vivo and in vitro. This review discusses how the mechanistic properties shared by the MSP machinery and actin-based motility systems lead to a "push-pull" mechanism for amoeboid cell motility in which cytoskeletal assembly and disassembly at opposite ends of the lamellipodium are associated with independent forces for protrusion of the leading edge and retraction of the cell body. PMID- 11061563 TI - Functional specificity of actin isoforms. AB - Actin, one of the main proteins of muscle and cytoskeleton, exists as a variety of highly conserved isoforms whose distribution in vertebrates is tissue specific. Synthesis of specific actin isoforms is accompanied by their subcellular compartmentalization, with both processes being regulated by factors of cell proliferation and differentiation. Actin isoforms cannot substitute for each other, and the high-level synthesis of exogenous actins leads to alterations in cell organization and morphology. This indicates that the highly conserved actins are functionally specialized for the tissues in which they predominate. The first goal of this review is to analyze the data on the polymerizability of actin isoforms to show that cytoskeleton isoactins form less stable polymers than skeletal muscle actin. This difference correlates with the dynamics of actin microfilaments versus the stability of myofibrillar systems. The three dimensional actin structure as well as progress in the analysis of conformational changes in both the actin monomer and the filament allows us to view the data on the structure and polymerization of isoactins in terms of structure-function relationships within the actin molecule. Most of the amino acid substitutions that distinguish actin isoforms are located apart from actin-actin contact sites in the polymer. We suggest that these substitutions can modulate the ability of actin monomers to form more or less stable polymers by long-range (allosteric) regulation of the contact sites. PMID- 11061564 TI - Cell biology of cardiac development. AB - Building a vertebrate heart is a complex task and involves several tissues, including the myocardium, endocardium, neural crest, and epicardium. Interactions between these tissues result in the changes in function and morphology (and also in the extracellular matrix, which serves as a substrate for morphological change) that are requisite for development of the heart. Some of the signaling pathways that mediate these changes have now been identified and several investigators are now filling in the missing pieces in these pathways in hopes of ultimately understanding the molecular mechanisms that govern healthy heart development. In addition, transcription factors that regulate various aspects of heart development have been identified. Transcription factors of the GATA and Nkx2 families are of particular importance for early specification of the heart field and for regulating expression of genes that encode proteins of the contractile apparatus. This chapter highlights some of the most significant discoveries made in the rapidly expanding field of heart development. PMID- 11061565 TI - Role of programmed cell death in development. AB - Programmed cell death (PCD) is an integral part of both animal and plant development. In animals, model systems such as Caenorhabditis elegans, Drosophila melanogaster, and mice have shown a general cell death profile of induction, caspase mediation, cell death, and phagocytosis. Tremendous strides have been made in cell death research in animals in the past decade. The ordering of the C. elegans genes Ced-3, 4 and 9, identification of caspase-activated DNase that degrades nuclear DNA during PCD, identification of signal transduction modules involving caspases as well as the caspase-independent pathway, and the involvement of mitochondria are some of the findings of immense value in understanding animal PCDs. Similarly, the caspase inactivation mechanisms of infecting viruses to stall host cell death give a new dimension to the viral infection process. However, plant cell death profiles provide an entirely different scenario. The presence of a cell wall that cannot be phagocytosed, absence of the hallmarks of animal PCDs such as DNA laddering, formation of apoptotic bodies, a cell-death-specific nuclease, a biochemical machinery of killer enzymes such as caspases all point to novel ways of cell elimination. Large gaps in our understanding of plant cell death have prompted speculative inferences and comparisons with animal cell death mechanisms. This paper deals with both animals and plants for a holistic view on cell death in eukaryotes. PMID- 11061566 TI - Reversible vacuolation of T-tubules in skeletal muscle: mechanisms and implications for cell biology. AB - The majority of investigations of the transverse tubules (T-system) of skeletal muscle have been devoted to their role in excitation-contraction coupling, with particular reference to contact with the sarcoplasmic reticulum and the mechanism of Ca2- release. By contrast, this review is concerned with structural and functional aspects of the vacuolation of T-tubules. It covers experimental procedures used in reversible vacuolation induced by the efflux-influx of glycerol and other small nonelectrolytes, sugars, and ions. The characteristics of the phenomenon, associated alterations in muscle function, and the swelling of analogous structures in nonmuscle cells are considered. Possible functions of reversible vacuolation in water balance, transport, membrane repair, muscle pathology, and fatigue are considered, and the potential application of reversible vacuolation in the transfection of skeletal muscle is discussed. In relation to the possible mechanisms involved in reversible vacuolation, particular attention is given to the dynamic and structural aspects of the opening and closing of T-tubules, the origin of vacuolar membranes, and the localized character of tubular swelling. PMID- 11061567 TI - Diagnostic value of the detection of t(14;18) chromosome translocation in malignant hematological and immunopathological diseases using polymerase chain reaction. AB - The majority of the t(14;18) chromosome translocations that occur in non-Hodgkin centroblastic-centrocytic follicular lymphoma can be detected by various methods. During the translocation process the bcl-2 gene located on chromosome 18 (18q21) is translocated to the JH region of the immunoglobulin gene of chromosome 14 (14q32). The most frequent type of bcl-2 translocations is the mbr type, whereas the immunoglobulin gene breaks mainly at the JH1-6 exons. About one of the 10(5) cells bearing the translocation can already be detected by using nested polymerase chain reaction (PCR). Eight patients suffering from follicular lymphoma were included in this study, which considered the usefulness of the PCR method. The results are in good agreement with those obtained by conventional diagnostic methods. Translocation can be detected, however, in patients with non malignant diseases such as Sjogren's syndrome (about 5% of the patients) and in a patient with Whipple disease. In addition, translocation was detected in lymphocytes of peripheral blood of a healthy donor. Since lymphomas are detected in patients with Sjogren's syndrome with a relative high frequency, an early diagnosis of the translocation could improve the treatment of the disease. Nevertheless, a diagnosis of lymphoma is valid only in cases of bone marrow translocation-positivity. PMID- 11061568 TI - Characterization of epstein-barr virus-infected mantle cell lymphoma lines. AB - It has been reported that Epstein-Barr virus (EBV) resides in resting B cells in vivo. However, an ideal in vitro system for studying EBV latent infection in vivo has not yet been established. In this study, a mantle cell lymphoma line, SP53, was successfully infected with a recombinant EBV containing a neomycin-resistant gene. The EBV-carrying SP53 cells were obtained by selection using G418. They expressed EBER-1, EBNAs, and LMP1; this expression pattern of the EBV genes was similar to that in a lymphoblastoid cell line (LCL). However, proliferation assay showed that the EBV-carrying SP53 cells have a doubling time of 73 h, compared with 57 h of SP53 cells. Transplantation of 10(8) SP53 cells to nude mice formed tumors in 4 of 10 mice inoculated, but the EBV-carrying SP53 cells did not. Unexpectedly, EBV infection reduced the proliferation and tumorigenicity of SP53 cells. However, the EBV-carrying SP53 cells showed higher resistance to apoptosis induced by serum starvation than did the SP53 cells. The inhibition of proliferation and the resistance to apoptosis induced in SP53 cells by EBV infection indicate that this cell line might to some extent provide a model of in vivo EBV reservoir cells. PMID- 11061569 TI - Calcineurin antagonists inhibit interferon-gamma production by downregulation of interleukin-18 in human mixed lymphocyte reactions. AB - Tacrolimus (FK-506) and cyclosporin A (CsA) are calcineurin antagonists used widely as T-cell immunosuppressants; however, their relative efficacy on the production of interleukin-18 (IL-18) remains undefined. We have examined the effects of FK-506 and CsA on the cytokine generation of human peripheral blood mononuclear cells (PBMCs) in mixed lymphocyte reaction (MLR) with lipopolysaccharide (LPS). We studied the levels of interleukin-18 (IL-18), IL-12, IL-10, IL-6, IL-2 and interferon-gamma (IFN-gamma) in the supernatant in allo-MLR by ELISA assay. Supernatant levels of IFN-gamma, IL-2, IL-6, IL-10 and IL-12 were detected 12 h after MLR and markedly increased thereafter. In contrast, production of IL-18 was detected at 12 h, reached a near maximum level at 24 h and decreased at 72 h. These results suggested that IFN-gamma production depended on IL-18, IL-12 and IL-2 in the early phase of MLR and depended mainly on IL-12 and IL-2 in the late phase. Both calcineurin antagonists inhibit the generation of IL-18, which plays a large role in allogeneic cell interactions, in macrophages and they also promote an equivalent down-regulation of T helper 1 (Th1) and Th2 responses in a concentration-dependent manner. About 90% of IFN gamma production induced by MLR was inhibited by an anti-IL-18 antibody, showing that IL-18 can trigger IFN-gamma production in MLR. These results suggest that dual signaling consisting of antigen-driven nuclear factor of activated T cells (NFAT) activation and LPS-mediated NF-kappaB activation is crucial for IL-18 production in macrophages, and that IL-18 can trigger IFN-gamma production in T cells by MLR. PMID- 11061570 TI - Fracture of the penis: treatment and complications. AB - Fracture of the penis is an unusual blunt traumatic injury of the erect penis. Twenty-five such cases, treated at the 2 main urological departments in the town of Varna between 1984 and 1999, were analyzed retrospectively. The average annual incidence in this series was 0.33 per 100,000 inhabitants. Most of the injuries occurred during vigorous sexual intercourse. Clinical diagnosis was mainly based on the patient's history and the physical findings. The diagnosis was verified by ultrasonography. In a few cases retrograde urethrography and cavernosography were used as additional diagnostic tools. Cavernosography was complicated by postprocedural priapism in 1 case. Unilateral tear of the corpora cavernosa was found in 24 cases. In 3 cases, partial (in 2) or complete (in 1) disruption of the corpus spongiosum and the urethra was found as well. The trauma was bilateral in 1 case in which 2/3 of the entire circumference of the penis was ruptured, including both corpora cavernosa and the urethra. Seventeen patients received immediate surgical repair. Most of them experienced an uneventful postoperative period with no serious consequences affecting their sexual function. Surgery was delayed in 6 and rejected in 2 cases. In all cases in which conservative (nonsurgical) management was the first treatment option, late complications (penile aneurysm, induration, penile curvature, erectile dysfunction) were observed and the final results were assessed as satisfactory or poor. We recommend immediate surgical treatment of all cases of penile fracture. Emergency surgical repair offers a chance for complete recovery and is the best method for providing a good functional prognosis. PMID- 11061571 TI - Biochemical modulation of 5-fluorouracil with a streptococcal preparation, OK 432, against murine colon-26 carcinoma. AB - Conventional therapy for colorectal carcinoma using 5-fluorouracil (5-FU) has shown limited antitumor action. The purpose of our study was to investigate synergistic antitumor effects of the streptococcal preparation of OK-432 and 5 FU, and to elucidate the mechanisms of interaction between the 2 agents in mice. Biochemical modulation of OK-432 and 5-FU were determined in vivo against colon 26 carcinoma. The concentration of 5-FU and its metabolites, and the activity of thymidylate synthase and thymidine kinase, respectively, were measured using cytosolic extracts of the tumors. Combination treatment with OK-432 produced a significant increase in intratumor 5-FU and 5-FU in RNA (F-RNA) concentrations, increased the thymidylate synthetase inhibition rate, and decreased thymidine kinase activity, as compared with the results observed in the control mice. These additive antitumor effects are obtained by use of the 2 agents; the mechanism of action is considered to be the suppression of both the de novo and the salvage pathway for DNA synthesis, along with the suppression of RNA synthesis. PMID- 11061572 TI - Anti-malarial activity of leaf-extract of hydrangea macrophylla, a common Japanese plant. AB - To find a new anti-malarial medicine derived from natural resources, we examined the leaves of 13 common Japanese plants in vitro. Among them, a leaf-extract of Hydrangea macrophylla, a common Japanese flower, inhibited the parasitic growth of Plasmodium falciparum. The IC50 of Hydrangea macrophylla leaf extract to Plasmodium falciparum was 0.18 microg/ml. The IC50 to NIH 3T3-3 cells, from a normal mouse cell line, was 7.2 microg/ml. Thus, selective toxicity was 40. For the in vivo test, we inoculated Plasmodium berghei, a rodent malaria parasite, to ddY mice and administered the leaf-extract of Hydrangea macrophylla (3.6 mg/0.2 ml) orally 3 times a day for 3 days. Malaria parasites did not appear in the blood of in the treated mice, but they did appear in the control group on day 3 or 4 after inoculation with the parasites. When leaf extract was administered to 5 mice 2 times a day for 3 days, malaria parasites did not appear in 4 of the mice but did appear in 1 mouse. In addition, the leaf-extract was administered orally 3 times a day for 3 days to Plasmodium berghei infected mice with a parasitemia of 2.7%. In the latter group, malaria parasites disappeared on day 3 after initiating the treatment, but they appeared again after day 5 or 6. Although we could not cure the mice entirely, we confirmed that the Hydrangea macrophylla leaf extract did contain an anti-malarial substance that can be administered orally. PMID- 11061573 TI - Endoscopic submucosal tumor biopsy using Stiegmann-Goff endoscopic ligator. AB - Endoscopic biopsy is an important tool for histological diagnosis of lesions residing in gastrointestinal tracts. However, it is less useful in submucosal lesions due to the existence of normal overlying mucosa. We developed a new and safe technique for the diagnosis of submucosal tumor using Stiegmann-Goff endoscopic ligator. After removing surface mucosa to expose submucosal tissue by this method, conventional secured histological diagnosis could be performed. To determine definitive histological diagnosis, this technique is useful as well as Endoscopic Ultrasound (EUS) with fine needle aspiration biopsy and other modalities. PMID- 11061574 TI - Induction of P-glycoprotein by rifampin increases intestinal secretion of talinolol in human beings: a new type of drug/drug interaction. AB - BACKGROUND: P-Glycoprotein is an efflux pump in many epithelial cells with excretory function. It has been demonstrated that rifampin (INN, rifampicin) induces P-glycoprotein, particularly in the gut wall. We therefore hypothesized that rifampin affects pharmacokinetics of the P-glycoprotein substrate talinolol, a beta1-blocker without appreciable metabolic disposition but intense intestinal secretion in human beings. METHODS: Pharmacokinetics of talinolol (a single dose of 30 mg administered intravenously or 100 mg administered orally for 7 days) and duodenal expression of the MDR1 gene product P-glycoprotein as assessed by reverse transcriptase-polymerase chain reaction of the MDR1-messenger ribonucleic acid, by immunohistochemistry and Western blot analysis were analyzed before and after coadministration of rifampin (600 mg per day for 9 days) in 8 male healthy volunteers (age 22 to 26 years). RESULTS: During rifampin treatment, the areas under the curve of intravenous and oral talinolol were significantly lower (21% and 35%; P < .05). Treatment with rifampin resulted in a significantly increased expression of duodenal P-glycoprotein content 4.2-fold (2.9, 6.51) (Western blot) and messenger RNA was increased in six of the eight volunteers. P-Glycoprotein expression in biopsy specimens of gut mucosa correlated significantly with the systemic clearance of intravenous talinolol (rs = 0.74; P < .001). CONCLUSIONS: Rifampin induces P-glycoprotein-mediated excretion of talinolol predominantly in the gut wall. Moreover, clearance of talinolol from the blood into the lumen of the gastrointestinal tract may be predicted by the individual intestinal P glycoprotein expression. Thus we describe a new type of steady-state drug interaction affecting compounds that are subject to transport rather than metabolism. PMID- 11061575 TI - In vivo and in vitro induction of human cytochrome P4503A4 by dexamethasone. AB - PURPOSE: The aims of these experiments were to determine the effect of a therapeutic regimen of dexamethasone on cytochrome P4503A4 (CYP3A4) activity in healthy volunteers; and the concentration-effect relationship between dexamethasone and CYP3A4 activity in primary human hepatocyte cultures. METHODS: The effect of dexamethasone (8 mg administered by mouth two times a day for 5 days) on CYP3A4 activity in 12 healthy volunteers was assessed with the erythromycin breath test and urinary ratio of dextromethorphan to 3 methoxymorphinan. Concentration-effect of dexamethasone on CYP3A4-dependent testosterone 6-beta-hydroxylation was determined in human hepatocytes treated with 2 to 250 micromol/L dexamethasone. RESULTS: The percent of erythromycin metabolized per hour increased from 2.20% +/- 0.60% (mean +/- SD) at baseline to 2.67% +/- 0.55% on day 5 of dexamethasone (mean increase in hepatic CYP3A4 activity 25.7% +/- 24.6%; P = .004). The mean urinary ratio of dextromethorphan to 3-methoxymorphinan was 28 (4.8 to 109) and 7 (1 to 23) at baseline and on day 5 of dexamethasone (mean decrease = 49%; P = .06). Substantial intersubject variability was observed in the extent of CYP3A4 induction. The extent of CYP3A4 induction was inversely correlated with baseline erythromycin breath test (r2 = 0.58). In hepatocytes, dexamethasone 2 to 250 micromol/L resulted in an average 1.7-fold to 6.9-fold increase in CYP3A4 activity, respectively. The extent of CYP3A4 induction with dexamethasone in hepatocyte preparations was inversely correlated with baseline activity (r2 = 0.59). CONCLUSIONS: These data demonstrate that dexamethasone at doses used clinically increased CYP3A4 activity with extensive intersubject variability and that the extent of CYP3A4 induction was, in part, predicted by the baseline activity of CYP3A4 in both healthy volunteers and human hepatocyte cultures. PMID- 11061576 TI - Steady-state pharmacokinetics of indinavir in cerebrospinal fluid and plasma among adults with human immunodeficiency virus type 1 infection. AB - To characterize steady-state indinavir pharmacokinetics in cerebrospinal fluid and plasma, 8 adults infected with human immunodeficiency virus underwent intensive cerebrospinal fluid sampling while receiving indinavir (800 mg every 8 hours) plus nucleoside reverse transcriptase inhibitors. Nine and 11 serial cerebrospinal fluid and plasma samples, respectively, were obtained from each subject. Free indinavir accounted for 94.3% of the drug in cerebrospinal fluid and 41.7% in plasma. Mean values of cerebrospinal fluid peak concentration, concentration at 8 hours, and area under the concentration-time profile calculated over the interval 0 to 8 hours [AUC(0-8)] for free indinavir were 294 nmol/L, 122 nmol/L, and 1616 nmol/L x h, respectively. The cerebrospinal fluid-to plasma AUC(0-8) ratio for free indinavir was 14.7% +/- 2.6% and did not correlate with indexes of blood-brain barrier integrity or intrathecal immune activation. Indinavir achieves levels in cerebrospinal fluid that should contribute to control of human immunodeficiency virus type 1 replication in this compartment. The cerebrospinal fluid-to-plasma AUC(0-8) ratio suggests clearance mechanisms in addition to passive diffusion across the blood-cerebrospinal fluid barrier, perhaps by P-glycoprotein-mediated efflux. PMID- 11061577 TI - Combined phenotypic assessment of CYP1A2, CYP2C19, CYP2D6, CYP3A, N acetyltransferase-2, and xanthine oxidase with the "Cooperstown cocktail". AB - BACKGROUND: Simultaneous administration of several probes enhances the utility of phenotyping, but poor specificity, side effects, and use of drugs not approved by the Food and Drug Administration limit the usefulness of prior phenotyping cocktails. OBJECTIVES: To evaluate potential drug-drug interactions associated with use of a cocktail of caffeine, omeprazole, dextromethorphan, and midazolam for simultaneous phenotyping of CYP1A2, CYP2C19, CYP2D6, CYP3A, N acetyltransferase-2, and xanthine oxidase. METHODS: Twelve subjects received caffeine + dextromethorphan, omeprazole, and midazolam (each alone), and a cocktail of caffeine + dextromethorphan + omeprazole + midazolam. Blood samples were collected at 120 minutes for omeprazole and 5/-hydroxyomeprazole, and at 0, 5, 30, 60, 120, 240, 300, and 360 minutes for midazolam. Twelve-hour urine samples were collected for analysis of dextromethorphan, caffeine, and metabolites. RESULTS: The median CYP1A2 metabolic ratio after administration of caffeine + dextromethorphan was not significantly different from that obtained with the cocktail (P = .84). Likewise, the median N-acetyltransferase-2, xanthine oxidase, and CYP2D6 metabolic ratios were not significantly different after cocktail administration (P = .977 for each N-acetyltransferase-2; P = .795 for xanthine oxidase; P = .75 for CYP2D6). The median CYP2C19 metabolic ratio after cocktail administration was not significantly different from that obtained after omeprazole administered alone (P = 1.000). Also, midazolam plasma clearance was not significantly different after cocktail administration compared with that after administration of midazolam alone (P = .708). The only side effect was sedation, which was associated with intravenous midazolam and occurred to a similar extent after both individual and cocktail phenotyping. CONCLUSIONS: These results indicate no pharmacokinetic or pharmacodynamic interactions that would limit the utility of this phenotyping cocktail for simultaneous measurement of the activity of multiple drug-metabolizing enzymes. PMID- 11061578 TI - Duration of effect of grapefruit juice on the pharmacokinetics of the CYP3A4 substrate simvastatin. AB - BACKGROUND: Grapefruit juice is a potent inhibitor of CYP3A4-mediated drug metabolism. We wanted to investigate how long the inhibitory effect of grapefruit juice lasts, with the CYP3A4 substrate simvastatin used as a model drug. METHODS: This crossover study consisted of 5 study days, during which 10 healthy volunteers ingested 40 mg simvastatin with water (control), with "high-dose" grapefruit juice (200 mL double-strength grapefruit juice three times a day for 3 days), or 1, 3, and 7 days after ingestion of "high-dose" grapefruit juice. For safety reasons, the study was performed in three parts to allow simvastatin-free days between the study days. Serum concentrations of simvastatin and simvastatin acid were measured by liquid chromatography-tandem mass spectrometry up to 12 hours. RESULTS: When simvastatin was taken with grapefruit juice, the mean peak serum concentration (Cmax) and the mean area under the serum concentration-time curve [AUC(0-infinity)] of simvastatin were increased 12.0-fold (P < .001) and 13.5-fold (P < .001), respectively, compared with control. When simvastatin was administered 24 hours after ingestion of the last dose of grapefruit juice, the Cmax and AUC(0-infinity) were increased 2.4-fold (P < .01) and 2.1-fold (P < .001), respectively, compared with control. When simvastatin was given 3 days after ingestion of grapefruit juice, the Cmax and AUC(0-infinity) were increased 1.5-fold (P = .12) and 1.4-fold (P = .09), respectively, compared with control. Seven days after ingestion of grapefruit juice, no differences in the Cmax or AUC(0-infinity) of simvastatin were seen. The mean Cmax and AUC(0-infinity) of simvastatin acid were increased 5.0-fold and 4.5-fold, respectively (P < .001), compared with control when simvastatin was taken with grapefruit juice and 1.7 fold (P < .01) when it was taken 24 hours after ingestion of grapefruit juice. After an interval of 3 or 7 days between ingestion of grapefruit juice and simvastatin, the pharmacokinetic variables of simvastatin acid did not differ significantly from those in the control phase. CONCLUSIONS: When simvastatin is taken 24 hours after ingestion of "high-dose" grapefruit juice, the effect of grapefruit juice on the AUC of simvastatin is only about 10% of the effect observed during concomitant intake of grapefruit juice and simvastatin. The interaction potential of even high amounts of grapefruit juice with CYP3A4 substrates dissipates within 3 to 7 days after ingestion of the last dose of grapefruit juice. PMID- 11061579 TI - Itraconazole alters the pharmacokinetics of atorvastatin to a greater extent than either cerivastatin or pravastatin. AB - BACKGROUND: 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors (statins) are metabolized by distinct pathways that may alter the extent of drug-drug interactions. Cerivastatin is metabolized by cytochrome P450 (CYP)3A4 and CYP2C8. Atorvastatin is metabolized solely by CYP3A4, and pravastatin metabolism is not well defined. Coadministration of higher doses of these statins with CYP3A4 inhibitors has the potential for eliciting adverse drug-drug interactions. OBJECTIVE: To determine the comparative effect of itraconazole, a potent CYP3A4 inhibitor, on the pharmacokinetics of cerivastatin, atorvastatin, and pravastatin. METHODS: In this single-site, randomized, three-way crossover, open labeled study, healthy subjects (n = 18) received single doses of cerivastatin 0.8 mg, atorvastatin 20 mg, or pravastatin 40 mg without and with itraconazole 200 mg. Pharmacokinetic parameters [AUC(0-infinity), AUC(0-tn), peak concentration (Cmax), time to reach Cmax (tmax), and half-life (t1/2)] were determined for parent statins and major metabolites. RESULTS: Concomitant cerivastatin/itraconazole treatment produced small elevations in the cerivastatin AUC(0-infinity), Cmax, and t1/2 (27%, 25%, and 19%, respectively; P < .05 versus cerivastatin alone). Itraconazole coadministration produced similar changes in pravastatin pharmacokinetics [AUC elevated 51% (P < .05 versus pravastatin alone), 24% (Cmax), and 23% (t1/2), respectively]. However, itraconazole dramatically increased atorvastatin AUC (150%), Cmax (38%), and t1/2 (30%) (P < .05). The elevation in atorvastatin AUC was significantly greater than that of cerivastatin (P < .005) or pravastatin (P < .005). CONCLUSION: Itraconazole markedly elevated atorvastatin plasma levels (2.5-fold) after 20 mg dosing, suggesting that concomitant itraconazole/atorvastatin therapy be carefully considered. Itraconazole produced modest elevations in the plasma levels of cerivastatin 0.8 mg or pravastatin 40 mg (1.3-fold and 1.5-fold, respectively), indicating that combination treatment with itraconazole with cerivastatin or pravastatin may be preferable. PMID- 11061580 TI - In vivo modulation of CYP enzymes by quinidine and rifampin. AB - BACKGROUND: The metabolism of drugs and other xenobiotics is mediated by enzymes whose activities can be modulated by different compounds. The activities of these modulators have the potential to be used to optimize drug action, prevent toxicity, or identify the enzymes involved in a reaction. This approach requires that selective agents be used for specific enzymes. However, selectivity of action has been poorly characterized in vivo. METHODS: This study investigated the effect of 3 and 28 days of treatment with quinidine (200 mg daily) and rifampin (INN, rifampicin) (600 mg daily) on the activities of four cytochrome P450 enzymes and N-acetyltransferase in 28 healthy young male volunteers divided into three groups with a cocktail of drug probes used, including caffeine, mephenytoin, debrisoquin (INN, debrisoquine), and dapsone. RESULTS: Quinidine selectively and almost completely inhibited the activity of CYP2D6 from day 3 through day 28 without affecting any other enzymes. Rifampin showed evidence of time-dependent induction of the activities of all measured oxidative routes of metabolism but decreased the acetylation ratio in fast acetylators. The quinidine/rifampin combination resulted in selective CYP2D6 inhibition and induction of all other enzymes evaluated over this time period, suggesting that predictable complex interactions occur with the drug combination. CONCLUSIONS: These observations illustrate the value of simultaneous assessment of the effect of modulators on the activities of multiple specific enzymes with the drug cocktail approach. PMID- 11061581 TI - The effect of hormone replacement therapy on CYP3A activity. AB - BACKGROUND: The effect of menopause and hormone replacement therapy on hepatic and intestinal wall CYP3A activity is poorly defined. This study was therefore designed to determine the effect of menopause and estrogen replacement therapy on hepatic and intestinal CYP3A activity with a specific CYP3A substrate, midazolam. METHODS: Twelve young women (27 +/- 5 years), 10 elderly women receiving estrogen replacement therapy (71 +/- 6 years), and 14 elderly women not receiving estrogen replacement therapy (71 +/- 5 years) received simultaneous intravenous (0.05 mg/kg over 30 minutes) and oral (3 to 4 mg of a stable isotope, 15N3-midazolam) doses of midazolam. Serum and urine samples were assayed for midazolam, 15N3 midazolam, and metabolites by use of gas chromatography-mass spectrometry. RESULTS: No significant (P > .05) differences were observed in systemic clearance and oral clearance between the three groups. Likewise, no differences were observed in oral, hepatic, or intestinal availability. A significant correlation was observed between oral and intestinal availability and not hepatic availability. CONCLUSION: Neither menopause nor menopause with estrogen replacement therapy altered intestinal or hepatic CYP3A activity relative to that in a control group of young women. PMID- 11061582 TI - Vascular effects of 5-HT1B/1D-receptor agonists in patients with migraine headaches. AB - OBJECTIVES: Second-generation triptans are believed to have fewer cardiovascular effects than sumatriptan. This was investigated in vivo by comparing the vascular effects of equipotent therapeutic dosages of selective 5-HT1B/1D-receptor agonists. METHODS: Sixteen patients with migraine headaches completed a double blind, placebo-controlled, four-way crossover study. With ultrasonography and applanation tonometry used 1.5 hours after the oral intake of sumatriptan (50 mg), rizatriptan (10 mg), zolmitriptan (2.5 mg), or placebo arterial vessel wall properties, blood flow and pressure waveforms were measured in common carotid, brachial, and temporal arteries. At the brachial artery, flow-induced vasodilation (an endothelium-dependent process) was evaluated, and blood pressures were recorded. RESULTS: Mean arterial pressure, 91 +/- 2 mm Hg after placebo, increased (P < .05) by 4% to 6% after administration of each triptan. Each active treatment decreased (P < .001) both brachial and carotid artery diameter. Isobaric compliance of the brachial artery, 0.077 +/- 0.010 mm2/kPa after placebo, decreased (P < .01) by 11% +/- 8%, 11% +/- 11%, and 23% +/- 7% after administration of sumatriptan, rizatriptan, and zolmitriptan, respectively. Isobaric compliance of the carotid artery was 1.31 +/- 0.10 mm2/kPa after placebo (no change). Zolmitriptan was the only triptan that decreased temporal artery diameter significantly (by 12% +/- 3%, P < .001). The resistance of the temporal artery vascular bed increased after administration of sumatriptan (by 26% +/- 11%, P < .05) and zolmitriptan (by 40% +/- 9%, P = .001). Flow-induced vasodilation was unaffected. CONCLUSIONS: Selective 5-HT1B/1D-receptor agonists induce vasoconstriction and decrease compliance of conduit arteries. These effects are more pronounced at muscular (temporal, brachial) compared with elastic (carotid) arteries. Resistance is only increased at the temporal artery vascular bed, suggesting cranioselectivity for resistance vessels. Endothelial function is not differently affected by any of the triptans tested. PMID- 11061583 TI - Simvastatin, capillary permeability, and acetylcholine-mediated vasomotion in atherosclerotic, hypercholesterolemic men. AB - OBJECTIVES: The aim of this study was to test the effect of high-dose simvastatin therapy on vascular permeability, a key variable in the atherogenic process, and endothelial-mediated vasodilator responses in patients with hypercholesterolemic atherosclerosis. METHODS: The transcapillary albumin escape rate (TERalb, the 1 hour decline rate of intravenous 125I-albumin, a measure of macromolecular permeability of capillary endothelium) and forearm vasodilatation (venous plethysmography) to intraarterial acetylcholine and sodium nitroprusside (7.5, 15, 30 microg/min and 0.8, 1.6, and 3.2 microg/min respectively, 5 minutes at each rate) to account for endothelium-dependent and independent mechanisms, were measured at baseline and after 1-month simvastatin (40 mg once daily) in 16 hypercholesterolemic (low-density lipoprotein cholesterol >130 mg/dL), atherosclerotic men. Thirteen healthy, untreated subjects were the controls. RESULTS: Baseline TERalb was higher and responsiveness to both acetylcholine and sodium nitroprusside was depressed in patients compared with controls. One-month high-dose simvastatin reduced low-density lipoprotein cholesterol by 39%, normalized TERalb, and improved local vasomotor responses to acetylcholine, without modifying those to sodium nitroprusside. Changes in TERalb and acetylcholine-mediated vasodilatation were dissociated and unrelated to lipid modifications. CONCLUSIONS: Low-density lipoprotein cholesterol reduction through 1 month of high-dose simvastatin normalized the exaggerated transvascular albumin leakage of patients with hypercholesterolemic atherosclerosis, perhaps by restoring an exaggerated endothelial permeability, apparently through mechanisms independent of circulating lipids. Improvements in acetylcholine-mediated vasomotion were also evident, but were dissociated from TERalb, demonstrating a heterogeneous behavior of the 2 indices of endothelial function in response to high-dose statin treatment. PMID- 11061584 TI - Paroxetine decreases platelet serotonin storage and platelet function in human beings. AB - BACKGROUND: Serotonin is a platelet agonist and potent vasoconstrictor that has recently received attention concerning its potential role in acute coronary artery thrombosis. Selective serotonin-reuptake inhibitors, such as paroxetine, are widely used antidepressant agents. We sought to characterize the potential inhibitory effect of paroxetine on platelet function. METHODS: Healthy male volunteers received 20 mg/d paroxetine for 2 weeks in a randomized, double-blind, placebo-controlled, two-way cross-over trial. RESULTS: Paroxetine decreased intraplatelet serotonin concentrations by -83% (P < .01). This inhibited platelet plug formation as reflected by a 31% prolongation of closure time measured with the platelet function analyzer-100 (P < .05). Furthermore, paroxetine lowered expression of the platelet activation marker CD63 in response to two different concentrations of thrombin receptor-activating peptide (P < .01). Plasma concentrations of prothrombin fragment, von Willebrand factor antigen, and circulating P-selectin remained unchanged in either period, indicating that paroxetine does not increase activation of coagulation, endothelium, or platelets in vivo, underlining a favorable safety profile. CONCLUSIONS: Paroxetine substantially decreases intraplatelet serotonin content and thereby reduces platelet plug formation under shear stress, and responsiveness to thrombin receptor activating peptide-induced platelet activation. Further studies will reveal whether these pharmacodynamic effects can be exploited for treatment of thrombotic artery disease. PMID- 11061585 TI - Hypotensive effect of calcium channel blockers is parallel with carbonic anhydrase I inhibition. AB - In this article we studied in vitro and in vivo the effect of calcium channel blockers (verapamil and amlodipine) on erythrocyte carbonic anhydrase I activity, on carbonic anhydrase I isolated from vascular smooth muscles, and on arterial blood pressure values in human beings and in animals. Our in vitro and in vivo results have shown that verapamil and amlodipine are strong inhibitors of carbonic anhydrase I both in erythrocytes (in human beings) and in vascular smooth muscles (in animals). In human beings calcium channel blockers reduce arterial blood pressure in subjects with hypertension and progressively reduce erythrocyte carbonic anhydrase I activity. We assume that verapamil and amlodipine possess a dual mechanism of action: the first mechanism consists of their action on calcium channels, and the second mechanism, proposed by us, shows that verapamil and amlodipine inhibit vascular smooth muscle carbonic anhydrase I activity with consecutive pH increase. The increase of pH might be an additional factor involved in intracellular calcium influx through calcium channels. This dual mechanism of action would bring new data regarding the hypotensive effect of verapamil and amlodipine, effects that might also be parallel and dependent on carbonic anhydrase I inhibition. PMID- 11061586 TI - Immunomodulatory effects of IL-12 in relation to the pineal endocrine function in metastatic cancer patients. AB - In addition to IL-2, IL-12 would constitute one of the most promising cytokines in the treatment of human neoplasms. IL-2 has been proven to induce in vitro and in vivo several evident changes in the secretion of cytokines and various other immunoinflammatory substances. In contrast, very little data are available about the immune effects of IL-12 in humans. The present study was carried out to investigate the in vivo immunoinflammatory effects of IL-12 by analyzing the secretions of neopterin, soluble IL-2 receptor (SIL-2R), tumor necrosis factor alpha (TNF), IL-2 and IL-6 in relation to the neuroendocrine function of the pineal gland, which is one of the most important organs involved in neuroimmunomodulation. Pineal endocrine function was investigated by evaluating the whole daily urinary excretion of the main catabolite of its hormone melatonin, 6-sulfatoxymelatonin (6-MTS). The study was performed on metastatic renal cell cancer patients. Each course of IL-12 consisted of 1.25 microg/ kg b.w. subcutaneously in the morning once a week for 3 consecutive weeks. The study evaluated 10 IL-12 courses. Mean serum levels of neopterin, SIL-2R and TNF significantly increased in response to IL-12, whereas no significant change occurred in IL-6 and IL-2 mean concentrations. Finally, 6-MTS urinary excretion was significantly reduced by IL-12 injection, particularly during the dark phase of the day. This preliminary study would suggest that IL-12 may induce important changes in the in vivo immunoinflammatory response. Moreover, IL-12 administration would suppress pineal endocrine activity, thus confirming its previously suggested involvement in the neuroimmunomodulatory processes. Because of the fundamental role of the pineal gland in neuroimmunomodulation, IL-12 induced immune variations could depend at least in part on its action at central neuroendocrine sites. PMID- 11061587 TI - Decrease of activated lymphocytes four and nine hours after a subcutaneous injection of a Viscum album L. extract in healthy volunteers. AB - The immunomodulatory substance, VaQuFrF (an aqueous extract of Viscum album L. of the oak tree) is used as an adjuvant and as monotherapy in the treatment of cancer and AIDS. After subcutaneous injection, there is a local inflammatory reaction at the injection site and systemic elevation of activated lymphocytes. The immunomodulatory effect of VaQuFrF in the first 24 h after subcutaneous injection on blood leukocyte and lymphocyte subpopulation was investigated. Because a significant natural circadian variation of these cellular parameters exist, the influence was studied in regard to this. In two groups of healthy volunteers, one group receiving VaQuFrF, the following parameters were measured every 2-3 h over a period of 24 h: leukocytes, band form, segmented and eosinophilic granulocytes, monocytes, total lymphocytes and CD4-, CD8-, CD3/25- and CD8/38-positive lymphocytes in count and percentage. In regard to the natural circadian variation 24 h after injection of VaQuFrF, a statistically significant fall in the absolute numbers and percentage of CD3/ 25- and CD8/38-positive lymphocytes was observed. Also, monocytes in percent and absolute numbers show a transient fall 6-9 h, lymphocytes only in absolute and CD4-positive lymphocytes only in percentage 2 h after injection. The results demonstrate that there is increased extravasation of (activated) lymphocytes and monocytes after subcutaneous injection of 1 mg VaQuFrF. PMID- 11061588 TI - The concept of an immune mechanism of chemical homeostasis and its importance in biology and medicine. AB - This paper outlines research that has led to the concept of the inevitable participation of the immune system in an organism's chemical homeostasis. This function of the immune system is tentatively named the 'immune mechanism of the chemical homeostasis' (IMCH). It is based on the theory of a permanent physiological synthesis of antibodies to endogenous biologically active substances. Minimal accumulation of biologically active substances as a result of the influence of different factors specifically activates the immune system in order to maintain its chemical homeostasis. The concept suggests the necessity of widening the notion of the range of the immune system's censorial functions. The concept explains the preexistence of immunocompetent cells preadapted to biologically active substances and autoantibodies specific to them; the natural clonality of the B lymphocyte pool; the polyclonal lymphocyte activation triggered by mitogens, foreign proteins, erythrocytes, and microbes, and tolerance to drugs. PMID- 11061589 TI - Autoantibody level modification in adult patients with idiopathic thrombocytopenic purpura following intravenous immunoglobulin treatment. AB - The aim of this study was to determine whether treatment of patients with immune thrombocytopenic purpura (ITP) with intravenous immunoglobulin (IVIg) is associated with a modification in the antiplatelet glycoprotein (GP) antibodies (Abs). Fourteen patients with ITP (11 females and 3 males, mean age 36.6 years, range 18-72) received one to four IVIg treatment courses. The preparation used was ISIVEN that was given in a dose of 2 g/kg body weight in a 5-day schedule and in monthly intervals. Levels of IgG, IgM and IgA isotypes of Abs to GPs IIb/IIIa and Ib/IX were measured before the treatment, and before and after each treatment course. Two patients did not respond to IVIg, 6 had a temporary response, 5 had a sustained response and 1 patient responded well to the treatment but was lost to follow-up. The patients had a high prevalence of serum Abs directed against GPs IIb/IIIa and Ib/IX before the treatment, and the mean IgG isotype levels of both Abs increased after each treatment course, and decreased again before the following course began. Whenever high Ab levels of either isotype (> 10 U/ml) were detected before the treatment, they were significantly decreased before the last treatment course. The elevated levels of IgG Abs to IIb/IIIa and Ib/IX after every course are probably a result of displacement of these Abs from Fc receptors by the IVIg, rather than of exogenous infusion of these Abs contained within the IVIg, whereas the decrease in high Ab levels after a few treatment courses results from the immunomodulatory effects of IVIg: suppression of Ab formation, and the presence of anti-idiotypes. PMID- 11061590 TI - Age-associated hematopoietic alterations in the spleen of tumor-bearing hosts. AB - The present investigation was carried out to study the effect of tumor growth on the spleen of an aged host. Dalton's lymphoma (DL), a spontaneous T cell lymphoma, was grown in mice of different age groups classified as young, adult or old on the basis of their reproductive status. Splenocytes obtained from normal and tumor-bearing young, adult and old mice were checked for an in vitro blastogenic response to concanavalin A (Con A), colony-forming ability and apoptosis. There was an enhanced apoptosis of splenocytes and a concomitant inhibition of splenocyte blastogenesis and their responsiveness to the mitogenic stimulus of Con A in aged mice. The counts of granulocyte macrophage- and macrophage-colony forming units were significantly enhanced in the spleen of tumor-bearing adult mice. It is proposed that the DL-growth-dependent increase in the size of the spleen in adult mice is due to an increased blastogenesis of splenocytes, which, however, may not be applicable in the case of old tumor bearing mice. The role of splenic macrophages in the regulation of the functions of the spleen by macrophage-derived NO is shown. PMID- 11061591 TI - CSF-1 (M-CSF) enhances the inflammatory response of fibronectin-primed macrophages: pathways involved in activation of the cytokine network. AB - We have previously reported that the priming of thioglycollate-elicited peritoneal macrophages (PMphi), as a representative population of mononuclear phagocytes (MNP), by macrophage-colony-stimulating factor (M-CSF or CSF-1) rendered these cells more susceptible to secondary stimulation by extracellular matrix (ECM) proteins, in particular fibronectin (FN), and that at least two beta1 integrins, VLA 4 (alpha4beta1 or CD49d) and VLA 5 (alpha5beta1 or CD49e), regulate IL-6 gene expression when PMphi come into contact with FN. In this report, we focused our attention on resident PMphi, as a more mature/differentiated MNP subpopulation. By using granulocyte-macrophage colony stimulating factor (GM-CSF)- and IL-6-knockout (null) mice, we demonstrated that the cooperative effect between CSF-1 and FN in IL-6 release was a result of a sequential stimulation of the GM-CSF, but not the TNF-alpha, gene via interaction with VLA 5. We also showed that regardless of the presence or absence of CSF-1 or FN, IL-6 inhibits GM-CSF and TNF-alpha gene expression in an autocrine manner. The observed effects were specific because CSF-1 enhanced VLA 5 expression and blocking FN-treated resident PMphi in vitro with VLA 5 monoclonal antibodies inhibited the IL-6 response. We found that treatment of resident PMphi with the protein kinase C inhibitor, staurosporine, and the activator, phorbol myristate acetate (PMA), resulted in marked modulation of either FN- or FN/CSF-1-induced cytokine release. An increased level of VLA 5 expression was observed in PMA treated resident PMphi. We concluded that in inflammatory processes, CSF-1 drives a number of pathways involved in the regulation of the expression of several genes and renders MNP highly susceptible to stimulation by ECM proteins that transform the MNP into secretory inflammatory cells. PMID- 11061592 TI - Expression of Fas and Fas ligand and apoptosis in tumor-associated lymphocytes and in tumor cells from malignant pleural effusions. AB - The CD95 (APO-1/Fas)-Fas ligand (FasL) system is an important mediator of antitumor T cell cytotoxicity. The aim of the current study was to assess its significance in human cancer. Malignant effusions were selected as an environment allowing direct cell-to-cell contact in a fluid phase. Malignant pleural effusions collected from 23 patients with metastatic carcinoma of the bronchus, ovary, stomach or breast were examined by means of flow cytometry. The expression ofFas and FasL, probed with the appropriate antibodies, apoptosis of tumor cells and the characteristics of tumor-associated lymphocytes (TAL) were determined by TUNEL reaction in malignant and nonmalignant (control) effusions. All malignant cells had partially or completely lost the expression of CD95 and expressed an elevated level of FasL. In contrast, TAL obtained from malignant pleural effusions demonstrated a marked decrease in the expression of surface FasL and an increase in surface-bound Fas. The percentage of apoptotic malignant cells was significantly decreased, as compared to TAL and lymphocytes from nonmalignant pleural effusions. There were also differences in the expression of Fas and FasL among mononuclear cells from malignant and nonmalignant pleural effusions. The ability of TAL from malignant pleural effusions to induce apoptosis of K562 cells was diminished, as compared to peripheral blood lymphocytes. Taken together, these data suggest that tumor cells in the microenvironment of malignant pleural effusions can evade immune attack by downregulation of the CD95 receptor and by killing lymphocytes through the expression of FasL. These results confirm earlier reports which showed that lymphocytes from a tumor microenvironment appear to have a depressed cytotoxic action. PMID- 11061593 TI - Clinical implications of natural killer cytotoxicity in patients with squamous cell carcinoma of the penis. AB - Impairment of natural cytotoxicity mediated by natural killer (NK) cells may play a role in the pathogenesis of penile carcinoma. The aim of this study was to examine the NK activity profile and its prognostic significance in patients with squamous cell carcinoma of the penis. The NK activity was measured in peripheral blood mononuclear cells (PBMCs) from 39 patients diagnosed histologically as having invasive squamous cell penile carcinoma and 4 patients with verrucous carcinoma of the penis. Of 39 patients with invasive squamous cell carcinoma, 4 had undergone previous penile amputation. According to the prognosis, the patients with invasive squamous cell carcinoma were divided into two groups: with metastasis and without metastasis. The patients were evaluated in relation to clinicopathologic variables using univariate analyses. NK cell activity was significantly decreased in all patients with penile carcinoma when compared with the control groups (p < 0.0001). There was no statistically significant difference between the groups with and without metastasis. We conclude that there is a decrease in NK activity in PBMCs from patients with penile carcinoma and that the presence of advanced disease or metastatic involvement is not responsible for this reduction. PMID- 11061594 TI - Apoptosis induced in T cells by human neuroblastoma cells: role of Fas ligand. AB - The CD95/CD95L (Fas/Fas ligand) receptor/ligand system plays an important role in regulation of cell survival and induction of a programmed cell death. It is also involved in regulation of effector phase of T and NK cell cytotoxicity, establishment of immune privilege sites, and tumor escape from immune recognition. In this study, we assessed expression of CD95L in tumors obtained from patients with neuroblastoma (NB) and in established NB cell lines. We measured the presence of intratumoral T cell infiltrates and T cell survival in tumor tissue samples. High levels of apoptosis were observed in tumor-associated lymphocytes as well as in Jurkat T cells cocultured with NB cells in vitro. T cell death was reduced after treatment of NB cells (in vitro) with antibody to FAS ligand (FasL). Overall, our data suggest that NB-induced apoptosis of Fas sensitive Jurkat T cells is mediated by functional FasL expressed on NB and Fas/FasL interaction may be responsible for the elimination of T cells in the NB microenvironment. PMID- 11061595 TI - Cancer mortality up, life expectancy down in Eastern Europe. PMID- 11061596 TI - Tobacco companies tried to destroy WHO. PMID- 11061597 TI - US administration halts cancer tests in Oklahoma. PMID- 11061598 TI - Breast cancer and the axilla: not entirely out of the labyrinth. PMID- 11061599 TI - Infections and human cancer. PMID- 11061600 TI - Anaplastic thyroid carcinoma: current diagnosis and treatment. AB - BACKGROUND: Anaplastic thyroid carcinoma (ATC), accounting for 5% to 15% of primary malignant thyroid neoplasms, is one of the most aggressive solid tumors in humans. Generally, it is rapidly fatal, with a mean survival of six months after diagnosis. Multimodality treatment with surgery and/or external beam radiotherapy and chemotherapy are of fundamental importance for local control of disease and to enhance survival. DESIGN: We evaluated consecutive patients with ATC observed at the Mayo Clinic from 1971 to 1993 and reviewed relevant articles published in major English-language medical journals. We used the MEDLINE database, selected bibliographies, and articles available in our personal files. RESULTS: ATC usually does not concentrate radioiodine or express thyroglobulin. It is essential to verify the diagnosis histologically because insular thyroid cancer, lymphomas, and medullary thyroid cancer are occasionally confused with undifferentiated neoplasms. Immunohistochemical study is helpful in establishing the diagnosis. Multimodal therapy and the development of effective systemic chemotherapeutic agents should result in improvements in survival, although no single agent has yet been identified. CONCLUSIONS: Aggressive multimodality treatment regimens show promise in improving local control in patients with ATC. However, survival rates remain low. Despite intense application of such therapy, no standardized successful treatment protocol has been established. PMID- 11061601 TI - Strategies to reduce the risk of virus-related cancers. AB - BACKGROUND: Experimental and epidemiological evidence has established an association between at least eight viruses and various cancer sites. Recent estimates (at least 10% of cancer worldwide) have revealed that viruses, together with tobacco and diet, account for the largest proportion of cancer in the world. RESULTS: Improvements in the detection of viruses and biomarkers of chronic infection have led to the identification of strong associations with cancer, particularly for human papillomavirus (HPV), hepatitis B virus (HBV), and human immunodeficiency virus (HIV). For some cancer viruses (e.g., HIV and hepatitis C virus, HCV), the spectrum of malignancies involved has still to be well defined. For HBV and HPV, vaccination aimed at cancer prevention is already a reality or a possibility. Whereas HBV vaccination already emerged as one of the most cost effective ways to reduce adult cancer mortality, for HPV vaccination some technical problems still await a solution. For other infectious agents (e.g., HCV, HIV) prospects for a vaccine are not immediate. CONCLUSIONS: In order to apply new knowledge on viruses to cancer prevention, large vaccination trials are warranted. These will have to be large (many thousands of people), prolonged (5 10 years), and match scientific excellence with a feasible design. Mistrust between scientists and the public will have to be prevented by means of absolute openness in scientific information and economical interests involved. PMID- 11061602 TI - Microsatellite alterations and TP53 mutations in plasma DNA of small-cell lung cancer patients: follow-up study and prognostic significance. AB - BACKGROUND: Small-cell lung cancer (SCLC), one of the major types of lung cancer, is associated with many different somatic molecular genetic changes. These alterations, observed in tumor DNA, have also been identified in the plasma DNA of patients. We undertook the present study to make a prospective investigation into the correlation between abnormal plasma DNA and patient survival. PATIENTS AND METHODS: Thirty-five patients with SCLC were selected after histological diagnosis. Polymorphic markers (ACTBP2, UT762 and AR) were chosen for their reported high rate of alterations in SCLC and analyzed in tumor tissue, normal blood cells and plasma DNA. Furthermore, we looked for mutations of the TP53 gene in tumor and plasma DNA. RESULTS: In 25 patients (71%) at least one molecular change precisely matching that of the primary tumor was detected in the plasma DNA. No difference in survival was observed between patients with aberrant plasma DNA and patients without plasma DNA alterations. However, patients with microsatellite modifications and TP53 mutations concomitantly, showed a significant difference (P = 0.02) in survival compared with patients bearing only one of these molecular changes. In 15 cases it was possible to find a correlation either between tumor response and disappearance of abnormal plasma DNA, or tumor progression and persistence of plasma DNA alterations. CONCLUSIONS: Free plasma DNA with molecular alterations is present to a high degree in plasma DNA of SCLC patients and may have a role as a prognostic factor. PMID- 11061603 TI - Acute hematologic toxicity and practicability of dose-intensified BEACOPP chemotherapy for advanced stage Hodgkin's disease. German Hodgkin's Lymphoma Study Group (GHSG). AB - BACKGROUND: Evidence is recently accumulating that the novel BEACOPP (bleomycin (B), etoposide (E), adriamycin (A), cyclophosphamide (C), vincristine (O), procarbazine (P), prednisone (P)) chemotherapy is a highly effective treatment for advanced stage Hodgkin's disease. Two dose variants of BEACOPP are currently tested in a phase III randomized multicenter trial of the GHSG. To enable more extensive testing of BEACOPP we characterized its practicability regarding schedule adherence, acute hematotoxicity and need for supportive treatment. PATIENTS AND METHODS: Data of 858 patients (6592 therapy cycles) from 184 participating institutions were evaluated. Planned total drug doses of the baseline variant (arm 1) were 80, 2400, 200, 5200, 11.2, 5600 and 4480 mg/m2 for B, E, A, C, O, P and P, respectively. Compared to arm 1, the doses of E, A and C in the dose-intensified variant (arm 2) were escalated by factor 2.0, 1.4, 1.92, respectively, using G-CSF assistance. Stepwise dose reductions were specified in case of dose-limiting toxicities. Both variants are given in eight three-weekly courses. RESULTS: Median dose adherence (dose actually given relative to planned arm 1 dose) in arm 1 was 1.0 for all drugs. Relative dose escalation of E, A, and C actually maintained in arm 2 was 1.83, 1.37 and 1.77 (medians), respectively, and 70% of patients maintained elevated dose levels throughout the entire treatment. Dose-limiting toxicities occurred in 25% of cycles in arm 2, most frequently due to leukocytopenia and thrombocytopenia. Time courses of leukocytes in arm 2 showed more severe but not more prolonged leukocytopenia compared with arm 1. WHO grades 3-4 infections were documented in 2.1% (arm 1) and 3.1% (arm 2) of all cycles. Erythrocytes were transfused in 61% (arm 1) and 28% (arm 2), platelets in < 1% (arm 1) and 6% (arm 2) of all cycles. CONCLUSIONS: Both BEACOPP schemes are practicable in a large multicenter setting. Despite increased hematotoxicity, moderate dose escalation is safe for the majority of the patients with G-CSF assistance and standard supportive treatment. PMID- 11061604 TI - Identification of prognostic subgroups among patients with metastatic 'IGCCCG poor-prognosis' germ-cell cancer: an explorative analysis using cart modeling. AB - OBJECTIVES: The IGCCCG classification has identified three prognostic groups of patients with metastatic germ-cell tumors. 'Poor prognosis' is based on primary tumor localization, the presence of visceral metastases, and/or high tumor-marker levels. The overall survival rate of these patients is about 45%-55%. The present analysis attempts to identify subsets of patients with a more or less favorable outcome among the 'poor-prognosis' group. PATIENTS AND METHODS: We retrospectively explored prognostic subgroups in 332 patients with 'IGCCCG' poor risk GCT using the classification-and-regression-tree model (CART). The following variables were included: primary tumor localization, presence of visceral or lung metastases, presence of an abdominal tumor, number of metastatic sites, serum levels of beta-HCG, AFP and LDH. All patients had been treated with cisplatin etoposide-based chemotherapy within controlled clinical trials between 1984 and 1997. RESULTS: PATIENT CHARACTERISTICS: gonadal/retroperitoneal (G/R) primary tumor 260 patients (78%), mediastinal primary tumor 72 patients (22%), visceral metastases 205 patients (62%) including 33 patients with CNS metastases, lung metastases 247 patients (74%), abdominal tumor 241 patients (72%), elevated AFP, beta-HCG or LDH levels 235 (71%), 253 (76%) and 275 (83%) of patients, respectively. Patients with primary mediastinal disease plus lung metastases exhibited the worst two-year PFS (28%), whereas patients with a primary G/R tumor and without visceral metastases showed the highest chance of two-year PFS (75%). The latter group of patients without visceral metastases and with a primary G/R tumor also had the most favourable two-year OS (84%). In contrast, patients with a primary mediastinal tumor and visceral metastases displayed the worst two-year OS (49%). CONCLUSIONS: Different prognostic subsets of patients can be identified among the group of 'poor-prognosis' GCT patients. The CART analysis model results in a hierarchy of prognostic factors which may allow to more precisely estimate the individual patient's prognosis. Identifying subgroups of 'very poor prognosis' among 'poor-prognosis' patients may allow to test for new treatment strategies in selected subgroups. PMID- 11061605 TI - Effect of single chemotherapeutic agents on the growing skeleton of the rat. AB - BACKGROUND: To establish the effect of chemotherapeutics on the growing skeleton, male Wistar rats were studied. DESIGN: Between the ages of 4 and 13 weeks the rats were given i.v. doxorubicin 15 mg/m2 body surface area (BSA), methotrexate 60 mg/m2 BSA or cisplatin 7.5 mg/m2 BSA. For each group of drug-treated rats there was a diet-matched control group that was injected with a placebo only. Rats fed ad libitum served as the basic control group for length and weight growth. Body weight and tibial length were measured weekly. Kidney and liver weight were determined at the end of the study. RESULTS: Weight gain and length growth were significantly decreased in the diet controlled groups (P < 0.05). Doxorubicin reduced length growth with 4.12 mm or 18% (P < 0.05). Methotrexate reduced length growth with 1.11 mm or 5% (P < 0.05). Length growth in the cisplatin treated rats did not differ from the diet controls. CONCLUSIONS: Doxorubicin and methotrexate decrease length growth in the rat tibia by, respectively, 18% and 5%. Cisplatin does not affect length growth. The decrease in growth might be a direct effect of doxorubicin and methotrexate on the tibial growth plate and metaphysis, but may be more pronounced due to the malnutrition. PMID- 11061606 TI - Octreotide acetate long-acting release in patients with metastatic neuroendocrine tumors pretreated with lanreotide. AB - BACKGROUND: In the present study we investigated the efficacy and tolerability of i.m. octreotide acetate (octreotide LAR) in patients with metastatic neuroendocrine tumors (NETs) previously treated and failed on i.m. lanreotide. PATIENTS AND METHODS: Fifteen patients (8 females, 7 males, median age 67 years, range 28-81 years) with metastatic NETs (8 endocrine pancreatic tumors, 7 midgut carcinoids) were enrolled in the study. All patients were in progressive disease (objective: 11 patients, symptomatic: 10 patients, biochemical: 11 patients) after treatment with slow release lanreotide, 30 mg every 14 days for a median time of 8 months (range 3-19 months). All patients had measurable disease; 12 patients had elevated serum and/or urine markers and 11 were symptomatic. Octreotide scintigraphy was positive in 13 of 15 patients. Octreotide LAR was administered as i.m. injection at the dose of 20 mg every four weeks until disease progression. RESULTS: An objective partial response (PR) was documented in one patient (7%), no change (NC) in six (40%), and progressive disease (PD) in eight patients (53%). The PR was observed in one patient with non-functioning endocrine pancreatic tumor with progressive liver and lymph node metastases after 16 months of i.m. lanreotide therapy. The median duration of disease stabilization was 7.5 months (range 6-12+ months). The overall biochemical response rate was 41%, including CRs (33%) and PRs (8%); biochemical responses were observed in carcinoids as well as in endocrine pancreatic tumors; the median duration of response was 5 months for CRs and 7.5 months for PRs. The overall symptomatic response rate was 82%. The median duration of response for diarrhoea, abdominal pain, or both was 6.5 months (range 3-12+ months). Improvement in performance status (PS) was obtained in 5 of 11 patients with PS of 1 at study entry. Median duration of octreotide LAR treatment was seven months (range 3-12+ months). No serious adverse events were reported; mild side effects were reported in 26% of patients. CONCLUSIONS: Octreotide LAR 20 mg shows significant efficacy in terms of objective response rate (PR + SD), biochemical and symptomatic control in patients with metastatic NETs of the GEP system pretreated and progressing on slow release lanreotide. PMID- 11061607 TI - Primary care physicians' knowledge and attitudes towards genetic testing for breast-ovarian cancer predisposition. AB - BACKGROUND: Primary health care providers are expected to be directly involved in the genetic testing for cancer susceptibility. This study assessed physicians' knowledge, attitude and perception of their role towards testing for hereditary breast ovarian cancer. DESIGN: A mail-in survey was sent to all general practitioners, internists, obstetrician-gynecologists and oncologists in private practice in Geneva county, Switzerland. Questions included socio-demographic variables, knowledge about hereditary breast ovarian cancer, attitude towards testing and assessment of their role in the pre- and post-test procedure. RESULTS: Two hundred fifty-nine (65%) of four hundred questionnaires were returned of which two hundred forty-three (61%) were analysed. Response rates were similar between specialties; women answered more frequently. The majority of the respondents (87%) approved of genetic susceptibility testing. The most common objection to testing was the absence of approved strategies for the prevention and detection of early breast cancer. Most physicians felt they had an active part to play in the pre-test procedure, the disclosure of results, and especially the consultants' long-term care and support (99%). Physicians correctly answered a third (32%) of the knowledge questions. The abstention rate for individual items ranged from 13% to 60%. Scores varied by specialty. Oncologists were more knowledgeable than gynecologists, internists and general practitioners. CONCLUSIONS: The majority of the primary care physicians in this study have a favourable attitude and are ready to play a prominent role in genetic counseling and testing for breast ovarian cancer predisposition. Defective knowledge scores, however, underline the need for targeted educational programs. PMID- 11061608 TI - Impact of nodal status on indication for adjuvant treatment in clinically node negative breast cancer. Istituto Nazionale per lo Studio e las Cura dei Tumori. AB - We addressed the problem of the need for axillary dissection in clinically node negative breast cancer by determining how the information provided by the dissection suggests a different treatment to that indicated by primary tumour characteristics and age alone. We examined retrospectively 260 cases of clinically node negative early breast cancer all of whom underwent breast surgery, radiotherapy and axillary dissection. We assigned adjuvant therapy according to accepted guidelines with and without consideration of pathological node status and compared the difference. Fifty-six patients had involved axillary nodes. There was no change in adjuvant chemotherapy indication for the 44 cases over 65 years. The change in indication for the remaining 216 cases was 18.5% to 6%, depending on whether none or all of the intermediate risk patients were originally assigned chemotherapy (all were originally assigned tamoxifen). Since the trend is for most intermediate risk patients is to be assigned adjuvant chemotherapy, and since tamoxifen is anyway considered effective therapy for low and intermediate risk patients, we conclude that the information provided by axillary dissection is probably not necessary if guidelines recommending wide application of systemic adjuvant chemotherapy are applied. Satisfactory prognostic information can be obtained by consideration of primary tumour characteristics, while new prognostic markers are likely to further refine prognostic precision in the near future. PMID- 11061609 TI - Phase II study of infusional chemotherapy with doxorubicin, vincristine and etoposide plus cyclophosphamide and prednisone (I-CHOPE) in resistant diffuse aggressive non-Hodgkin's lymphoma: CALGB 9255. Cancer and Leukemia Group B. AB - BACKGROUND: Patients with resistant diffuse aggressive non-Hodgkin's lymphoma (DA NHL) have a poor prognosis. Studies have suggested infusional therapy may be beneficial. PATIENTS AND METHODS: This trial used an infusional regimen called I CHOPE in resistant patients who had previously received only bolus CHOPE or CHOP regimen. Resistance was defined as: a) primary refractory disease, b) progression on therapy, c) partial response, d) complete remission lasting less than one year. Eligibility criteria included a diagnosis of DA-NHL (IWF E-H), no prior irradiation and adequate organ function. RESULTS: Thirty-seven patients were entered and twenty-nine were eligible. Reasons for ineligibility were incorrect histology (5) and other (3). The median age was 57 years (range 29-81) with 21 males. The performance status scores were: 0 (12 patients); 1 (9 patients); 2 (8 patients). Prior therapy consisted of standard CHOP (26 patients), bolus CHOPE (2 patients), high dose CHOP (1 patient). Therapy consisted of a 120 hour continuous intravenous infusion of doxorubicin 10 mg/m2/day, vincristine 0.28 mg/m2/day (maximum 0.4 mg/day), and etoposide 48 mg/m2/day. Cyclophosphamide 750 mg/m2 was given as an i.v. bolus day 6 and prednisone was given at 100 mg/day p.o. on days 1-5. G-CSF was allowed for myelosuppression. The overall response rate was 48% (CR 17%; PR 31%). Freedom from progression was 24% at six months and 8% at one year. Survival was 69% at six months and 40% at one year. In an exploratory analysis a prior CR or PR predicted response to I-CHOPE. Twelve of sixteen patients who had a CR/PR on previous therapy responded while two of thirteen who had no prior response, responded to I-CHOPE (P = 0.003). The toxicity was tolerable with grade 3-4 hematologic toxicity being leucopenia 94% and thrombocytopenia 41%. The grade 3-4 non-hematologic toxicities were infection in 28%, phlebitis in 11%, and stomatitis in 15%. CONCLUSIONS: I-CHOPE can induce responses in this group of patients with a poor prognosis, but most were seen in those who had previously had a response to bolus chemotherapy. PMID- 11061610 TI - Multimodality therapy in inflammatory breast cancer: is there a place for surgery? AB - BACKGROUND: In many centres surgery is used as part of a combined modality approach to the treatment of inflammatory breast cancer (IBC). Nevertheless, its value is controversial given the high risk of metastatic relapse and poor overall prognosis. We have reviewed patients with true IBC prospectively treated at the Royal Marsden Hospital in chemotherapy trials to assess further the role of surgery as part of combined modality treatment. PATIENTS AND METHODS: Fifty-four patients who had responsive or stable disease to primary chemotherapy went on to have either radiotherapy alone (n = 35) or surgery plus radiotherapy (n = 19); the decision on surgery was based partly on clinician preference and partly on clinical response. RESULTS: The 35 patients undergoing radiotherapy alone had a median progression-free survival (PFS) of 16 months and median overall survival (OS) of 35 months. Twenty-four patients (69%) have relapsed with a total of twelve (34%) relapsing locally. In comparison, the 19 patients receiving both surgery and radiotherapy had a PFS of 20 months, and a median OS of 35 months. Fifteen patients (79%) have relapsed, eight (42%) of these locally. None of these differences were statistically significant. CONCLUSIONS: These results do not suggest a clinical advantage for surgery in addition to chemotherapy and radiotherapy for patients with IBC. They support the need for a prospective randomised trial to address this question. PMID- 11061611 TI - Vinorelbine and cisplatin in metastatic breast cancer patients previously treated with anthracyclines. AB - PURPOSE: To assess the antitumor efficacy and safety of a vinorelbine and cisplatin combination in patients with metastatic breast cancer previously treated with anthracyclines. PATIENTS AND METHODS: Fifty-three patients with assessable metastatic breast cancer with previous exposure to anthracyclines (adjuvant n = 6, palliative n = 47) were studied. Cisplatin 75 mg/m2 on day 1 was given followed by 25 mg/m2 vinorelbine (VNR) on days 1 + 8, in a five-min i.v. infusion. Courses were repeated every three weeks on an outpatient basis. Treatment continued until disease progression, excess toxicity or patient refusal. Patients were classified according to their response to anthracyclines: anthracycline refractory patients were patients who had never responded under an anthracycline regimen. Anthracycline resistant patients were either metastatic patients who progressed within four months of completing anthracycline-based chemotherapy or patients who progressed within six months of completion of an anthracycline adjuvant treatment. Patients who progressed four months after the end of an anthracycline regimen in metastatic setting or six months after the end of an anthracycline regimen in adjuvant setting were considered as patients previously treated with anthracyclines and were called 'relapsed'. RESULTS: Four patients (8%) achieved a complete response (CR) and twenty-two patients (41%) achieved a partial response (PR) with an overall response rate (OR) of 49% (95% confidence interval (CI): 35-63). Stable disease (SD) was observed in five patients (9%), twenty-two patients had progressive disease (PD). Responses according to previous sensitivity to anthracycline were as follow: 5 refractory patients achieved a PR from 14 patients (36%). Seven of sixteen resistant patients responded (44%), six with PR and one with CR. Among 23 'relapsed' patients, 14 responses were observed (61%), with 3 CR and 11 PR. There was no statistical difference in RR among the three groups. The median duration of response for all patients was 7 months, the median time to progression (TTP) 5 months and median overall survival 12 months. All patients were assessed for toxicity. The main toxicity was neutropenia grade 3 and 4 in 49% of patients. Febrile neutropenia requiring hospitalization was uncommon (2 patients). There were no treatment related deaths. Despite potential overlapping neurologic toxicities of the two drugs, only eight patients (15%) developed neuropathy, which was, however, mild (grades 1 and 2). CONCLUSIONS: This cisplatin VNR regimen is well tolerated and active in patients who failed anthracyclines. The response rate, TTP and survival data are encouraging and indicate that cisplatin VNR may have a place as second-line treatment alternative to taxanes or other less active regimens. If these results can be verified in multi-institution trials, this combination of drugs would merit investigation as first-line therapy in this patient population. PMID- 11061612 TI - Phase II trial of gemcitabine in patients with previously untreated metastatic cancer of the esophagus or gastroesophageal junction. AB - BACKGROUND: There were approximately 12,500 cases of esophageal carcinoma diagnosed in the US in 1992 and 12,200 deaths. The impact of chemotherapy on patients with metastatic disease is marginal with a median survival of only five months. Gemcitabine (LY188011,2,2,-difluorodeoxycytidine: dFdC), an analog of cytosine arabinoside (ara-C), is a pyrimidine antimetabolite. Gemcitabine has shown interesting clinical activity in initial phase II clinical trials in a variety of malignancies, including the aerodigestive malignancies, squamous-cell carcinoma of the head/neck and both non-small-cell and small-cell lung cancer. PATIENTS AND METHODS: A total of 21 patients with chemotherapy-naive metastatic esophageal carcinoma were entered. Nineteen patients were evaluable for toxicity and seventeen patients were evaluable for response. Gemcitabine was administered intravenously at 1250 mg/m2 over 30-60 minutes on days 1, 8, and 15 followed by 1 week of rest. This four-week schedule defined a cycle of treatment. Patients may have received a maximum of six cycles. RESULTS: Gemcitabine was well tolerated with minimal non-hematologic toxicity and grade 3-4 anemia, granulocytopenia, and thrombocytopenia occurring in 10.5%, 21%, and 0% of patients, respectively. No responses were seen in the seventeen evaluable patients. CONCLUSIONS: At the dose and schedule studied it would appear that gemcitabine has no activity in patients with chemotherapy-naive esophageal carcinoma. PMID- 11061613 TI - A phase II study of the 5-lipoxygenase inhibitor, CV6504, in advanced pancreatic cancer: correlation of clinical data with pharmacokinetic and pharmacodynamic endpoints. AB - PURPOSE: Primary objective was to determine response rate of patients with advanced pancreatic cancer to a novel lipoxygenase and thromboxane A2 synthetase inhibitor (CV6504); secondary objectives included estimation of pharmacokinetics of CV6504, target-enzyme inhibition, safety and tolerance, quality of life and survival. PATIENTS AND METHODS: Thirty-one patients with advanced pancreatic cancer were planned to receive CV6504, 100 mg TDS, orally for three months, at which point CT scans were performed to assess therapeutic response rates. Steady state concentrations of CV6504 and thromboxane B2 (an indirect measure of thromboxane A2 synthetase (TA2S) inhibition) were made. Of the 31 patients entered into the study, 23 were considered fully evaluable for response. RESULTS: The drug was well tolerated with few side effects; no partial or complete responses were seen, but 10 patients had stable disease at 3 months; quality of life was maintained during therapy; mean CV6504 steady state plasma concentrations of 14 +/- 6 ng/ml resulting in 75 +/- 18% inhibition of TA2S were achieved; median-survival time for all patients considered eligible for assessment of efficacy was 36.6 weeks after the initial dose of study medication. The actuarial one-year survival was approximately 25%. CONCLUSION: CV6504 inhibits its target enzyme in vivo, maintains stable disease in 32% of evaluable patients and is well tolerated. PMID- 11061614 TI - Aspirin and ovarian cancer: an Italian case-control study. AB - BACKGROUND: A role for inflammation, and consequently of non-steroidal anti inflammatory drugs, in ovarian carcinogenesis has been proposed, but epidemiological evidence is scanty. PATIENTS AND METHODS: Data were derived from a hospital-based case-control study conducted in Italy between 1992 and 1999. Cases were 749 women, aged 18-80 years (median age 56 years), with incident, histologically confirmed ovarian cancer. Controls were 898 non-hysterectomized women, aged 17-80 years (median age 58 years), admitted to hospital for acute conditions, unrelated to risk factors for ovarian cancer. RESULTS: The multivariate odds ratio (OR) was 0.93 (95% confidence interval (95% CI): 0.53 1.62) for regular aspirin use for more than six months, 1.38 (95% CI: 0.57-3.36) for current use and 0.72 (95% CI: 0.35-1.47) for former use. The OR was not significantly different from unity for duration of use, age at starting use, indication (analgesia or cardiovascular prevention), and in women aged < 60 and > or = 60 years at ovarian cancer diagnosis. CONCLUSIONS: This study provides little support for the hypothesis that aspirin may reduce the risk of ovarian cancer. PMID- 11061615 TI - A randomised, prospective, phase III clinical trial of primary bleomycin, ifosfamide and cisplatin (BIP) chemotherapy followed by radiotherapy versus radiotherapy alone in inoperable cancer of the cervix. AB - BACKGROUND: Phase II studies have shown primary (neo-adjuvant) chemotherapy with bleomycin, ifosfamide and cisplatin (BIP) is active against inoperable cervical cancer. We present here results of a randomised phase III multicentre trial comparing radical radiotherapy with neo-adjuvant BIP chemotherapy followed by radical radiotherapy in patients with inoperable cervical cancer, designed to discover whether this combination might improve survival. PATIENTS AND METHODS: Patients with inoperable cervical carcinoma were randomised to pelvic radiotherapy alone [RT] or two to three cycles of bleomycin 30 units/24-hour infusion, ifosfamide 5 g/m2/24 hours, and cisplatin 50 mg/m2) chemotherapy followed by pelvic radiotherapy (BIP + RT). Randomisation was stratified by stage and radiotherapy centre. RESULTS: One hundred seventy-two eligible women were randomised into this trial; eighty-six to RT and eighty-six to BIP + RT. A total of 190 cycles of chemotherapy were given. Median follow-up for the 47 patients still alive is 9 years with a minimum follow-up of 3 years. Complete or partial response occurred in 51 of 86 (59%) of those randomised to RT and 60 of 86 (69%) of those randomised to BIP + RT. The difference between response rates does not reach statistical significance (chi2 = 2.06, P = 0.15). Median survival is two years with an actuarial survival at five years of 32% (95% confidence interval (95% CI): 25%-39%). There is no significant difference between the treatment groups (chi2log-rank = 0.11, P = 0.74). CONCLUSIONS: This study does not show any survival benefit from the use of neo-adjuvant BIP chemotherapy in advanced cervical cancer. PMID- 11061616 TI - GM-CSF with biochemotherapy (cisplatin, DTIC, tamoxifen, IL-2 and interferon alpha): a phase I trial in melanoma. AB - BACKGROUND: Ineffective tumour antigen processing is recognised as an important cause of failure of immunotherapy in melanoma. GM-CSF may augment the cytotoxic lymphocyte response by activating antigen-presenting cells. This study evaluates a schedule combining GM-CSF with biochemotherapy. PATIENTS AND METHODS: Nineteen patients with advanced malignant melanoma received cisplatin (25 mg/m2 days 1-3). dacarbazine (220 mg/m2 days 1-3), interleukin-2 (9 MIU/m2/24 h) and interferon alpha2b (5 MIU/m2) both days 6-10 and days 17-21, and tamoxifen 40 mg/day continuously. Subcutaneous GM-CSF was given in escalating doses to three cohorts: 1) 450 microg/m2 days 4-5 and 15-16; 2) as 1) plus 225 microg/m2 days 6-10 and 17 21; 3) 450 microg/m2 days 4-10 and 15-21. Each cycle was 28 days. RESULTS: Constitutional side effects were the major non-haematological toxicity and lymphopaenia the main haematological toxicity. Six patients responded (32%, 95% confidence interval: 13%-57%), two patients had complete remission. There was an apparent trend for increasing responses with increasing GM-CSF dose; zero of six responses in cohort 1, two of seven in cohort 2 and three of six in cohort 3 (P = 0.016). Median overall survival was 6.2 months. Increasing GM-CSF doses significantly increased serum concentrations of neopterin and TNF-alpha. CONCLUSIONS: The combination of GM-CSF with biochemotherapy is feasible and there appears to be a dose-response relationship with GM-CSF in terms of host immunological response, and possibly clinical efficacy. PMID- 11061617 TI - Distal ischemic changes related to combination chemotherapy with cisplatin and gemcitabine: description of four cases. PMID- 11061618 TI - Hepatocellular carcinoma after long-term tamoxifen therapy. AB - We describe a case of hepatocellular carcinoma (HCC) after long term tamoxifen therapy in a 71-year-old woman. The patient was prescribed tamoxifen for 12 years following right mastectomy and axillary node clearance for breast carcinoma in 1985. In 1997, she complained of abdominal pain and fullness. An abdominal ultrasound scan showed lesions in the right lobe of liver which were thought to be metastases. However, a biopsy showed primary HCC. Studies in rats suggest that tamoxifen is involved in hepatic carcinogenesis but studies in humans have failed to show any increased risk. However, these studies followed up patients for less than five years. An increased risk of HCC may not become apparent until after a decade or more of tamoxifen therapy. In addition, HCC in tamoxifen treated patients may be under-reported since there may be reluctance to biopsy liver tumours which are assumed to be secondary carcinoma of the breast. PMID- 11061619 TI - Inflammatory demyelinating polyneuropathy: a complication of immunotherapy in malignant melanoma. AB - Paraneoplastic syndromes (PNS) involving the central nervous system are a rare manifestation of malignant disease. As they commonly precede the diagnosis of malignancy their acute manifestations do not often present themselves to oncologists in the first instance. It is currently believed that most, if not all, neurological PNS are autoimmune in nature. Proteins expressed ectopically on the surface of tumour cells generate an immune response which cross-reacts with the same, or similar, proteins in the nervous system resulting in damage. This can involve a single cell type of the nervous system whilst in other cases the impairment is more widespread. The following report is of a case of chronic inflammatory demyelinating polyneuropathy (CIDP) occurring in metastatic malignant melanoma, following treatment with interferon-alpha. We review the current literature on this rare association and speculate on its pathogenesis, and the implications for future therapeutic strategies in melanoma targeting tumour antigens. PMID- 11061620 TI - Access to hospital care, clinical stage and survival from colorectal cancer according to socio-economic status. PMID- 11061621 TI - Phase I and pharmacological study of increased dose oral topotecan in combination with intravenous cisplatin. PMID- 11061622 TI - Is there a range of time for initiation of adjuvant chemotherapy in patients with malignancy? PMID- 11061623 TI - Virulence profiles and other biological characters in water isolated Aeromonas hydrophila. AB - Thirty water isolates of A. hydrophila were tested for potential virulence profiles, antibiotic resistance and Bacteriocin-Like Substances (BLS) production. Cytotoxic activity was present in all strains tested, 87% were hemolytic and 70% adhesive. Lysine decarboxylase reactions (LDC) positivity was correlated with virulence factors: 100% versus cytotoxicity, 84% versus adherence, 76% versus hemolytic activity. The correlation was also present in the LDC-negative strains. Hemolytic and cytotoxic activities were frequently associated: high cytotoxicity, corresponding to high hemolytic activity and vice versa. The in vitro susceptibility of A. hydrophila to 28 antibacterial agents showed that cefotaxime was the most active beta-lactam antibiotic, and Cefuroxime inhibited 90% of the strains. Isolates were resistant to Penicillin G, Ampicillin, Carbenicillin, Amoxicillin, Cephalotin and Cefaclor. Tetracycline, Chloramphenicol, Nitrofurantoine, the quinolones and the aminoglycosides (except Streptomycin) were consistently active. BLS production never emerged against closely-related microorganisms. On the contrary A. hydrophila presented a heteroinhibitory activity against non-taxonomically related genera such as Listeria spp. (L. seeligeri NCTC 11856, L. welshimeri NCTC 11857, L. ivanovii NCTC 11846) and S. aureus ATCC 25923. Although a large number of strains showed virulence determinants together with other biological characters such as antibiotic resistance and BLS production, it was not possible to relate these factors to the observed plasmids. PMID- 11061624 TI - Multicenter evaluation of the new HIV DUO assay for simultaneous detection of HIV antibodies and p24 antigen. AB - A multicenter survey was performed to evaluate a new semi-automated human immunodeficiency virus fourth generation antibodies and antigen simultaneous assay. This assay showed a sensitivity of 100% and specificity of 99.6% among sera obtained from hospitalized patients or blood donors. Sera obtained from commercially available as well as in-house seroconversions were tested showing that HIV DUO is able to reveal an infected state in 11 out of 14 cases earlier than conventional tests. This new assay improves old test performances in terms of sensitivity, maintaining specificity at very high levels. PMID- 11061625 TI - Results from the first computerised Italian surveillance of human Salmonella isolates. The Italian SALM-NET Working Group. AB - We report all the first computerised data collected in Italy for the surveillance of Salmonella isolates. Primarily, within the wide framework of the European Community Human Salmonella Surveillance Project (SALM-NET), we report data on the most commonly isolated serotypes in Italy from January 1994 to December 1996. In addition, we report all computerised data historically collected by some Italian regions regarding the period 1980-1993. Total data included in the Italian SALM NET data base account for 59,336 Salmonella isolates. In the list of the most frequent isolates starting from 1989, S. Enteritidis always ranked first, followed by S. Typhimurium. PMID- 11061626 TI - Comparative evaluation of two commercially available antigen enzyme immunoassays (EIA) for the detection of Legionella pneumophila urinary antigen in frozen non concentrated urine samples. AB - We evaluated two commercial enzyme immunoassay kits, Binax EIA (for detection of soluble antigen of Legionella pneumophila serogroup 1) and Biotest EIA (for detection of antigens of Legionella pneumophila serogroups and other Legionella spp.) in order to introduce this test routinely for the diagnosis of Legionnaires' disease (LD) in our Laboratory. Frozen non-concentrated urine samples belonging to 45 patients with and without LD were tested. The sensitivity of Binax EIA and Biotest EIA was 47.4% and 42.1% respectively, the specificity was 95% by both tests. Biotest did not detect antigen from a patient with culture proven infection of L. pneumophila serogroup 6. The detection of urinary antigen by both EIA tests is a useful tool for rapid diagnosis of LD, especially when samples are unavailable for culture; the sensitivity may be increased if the assay is performed on unfrozen and concentrated samples. PMID- 11061627 TI - Evaluation of OptiMAL Assay test to detect imported malaria in Italy. AB - This study evaluated a newly developed rapid malaria diagnostic test, OptiMAL Assay, to detect "Plasmodium falciparum malaria" and "non Plasmodium falciparum malaria" in blood samples from 139 individuals with a presumptive clinical diagnosis of imported malaria in Italy. OptiMAL Assay utilizes a dipstick coated with monoclonal antibodies against the intracellular metabolic enzyme, plasmodium Lactate Dehydrogenase (pLDH) present in and released from parasite-infected erythrocytes. Blood samples from 56 cases out of 139 were found "Plasmodium falciparum malaria" positive by microscopy; with these samples OptiMAL Assay and the ParaSight-F test, which is a kit detecting the P. falciparum histidin-rich protein 2 (HRP-2), showed an overall sensitivity of 83% and 94%, respectively, in comparison with microscopy. Parasitemia levels tested in the 56 P. falciparum positive blood samples by microscopy ranged from <0.004% to 20%. A correlation between sensitivity and parasitemia was evident and OptiMAL Assay and ParaSight-F test were more sensitive (96-100%; 100%) with samples with 0.1%-20% levels of parasitemia, while proved less sensitive (0-44%; 50-88%) with <0.004-0.01% levels of parasitemia. PMID- 11061628 TI - Assessment of a monoclonal antibody-based competitive enzyme linked immunosorbent assay (cELISA) for diagnosis of brucellosis in infected and Rev. 1 vaccinated sheep and goats. AB - In this study a cELISA for the diagnosis of brucellosis due to B. melitensis in sheep and goats was evaluated and its capability of discriminating vaccinated from infected animals was assessed. Information is provided indicating that the cELISA has a diagnostic sensitivity (99.4%) and specificity (98.9%) in sheep and goats comparable to that of many standard indirect ELISA methods. In addition, the test proved able to distinguish between vaccinated and infected animals with an accuracy of up to 90% and results reproducibility of 93%. It was concluded that the cELISA could be useful for differentiation of Rev.1 vaccinated and naturally infected sheep and goats. PMID- 11061629 TI - Biofilm formation on the surface of ceramic tiles. AB - The aim of the study was to investigate the formation of biofilm on the surface of ceramic tiles, widely present in public and private buildings, using six parallel flow chambers. Our flow system was conceived and made to compare biofilm results by parallel distributed rectangular tiles. The tiles, divided into two identical A and B sections, were placed within the flow chambers. Biofilm formation was performed after 72 h and was quantified by viable counts of bacteria. Average viable counts ranged from 1.1x10(7) to 7.3x10(7) cfu cm(-2) and from 1.1x10(7) to 5.8x10(7) cfu cm(-2) respectively for biofilm A and B sections. As statistical analysis does not show significant differences, we can conclude that biofilms obtained were so similar to each other that they confirmed the system reproducibility. Our next step will be to use our system to study Legionella pneumophila and to evaluate the efficacy of antibacterial agents. PMID- 11061630 TI - Influence of an extended incubation period on values of minimum inhibitory concentrations of antibiotics in Stenotrophomonas maltophilia clinical strains. AB - In 106 Stenotrophomonas maltophilia clinical strains the susceptibility to 19 kinds of antibiotics was tested by the broth dilution micromethod at 24 h and 48 h incubation. Isolated strains demonstrated the lowest frequency of resistance to cotrimoxazole (7.5% of resistant strains at 24 h incubation and 18.9% at 48 h), ofloxacin (13.2% and 30.2%), ciprofloxacin (19.8% and 50.9%) and to cefoperazone/sulbactam (20.8% and 37.7%). The smallest growth of the number of resistant strains after extended incubation was recorded in gentamicin (by 10.4%), ceftazidime (by 11.3%) and cotrimoxazole (by 11.4%). On the contrary, the largest growth of resistance was demonstrated in cefoperazone and ciprofloxacin (by 31.1%). Average values of the growth of minimum inhibitory concentrations (MICs) were lowest in ciprofloxacin and ofloxacin (2.3 times) and highest in piperacillin/tazobactam (4.5 times) and piperacillin (5.0 times). As far as the stability of MIC is concerned, the largest occurrence of strains with the MIC growth doubled as a maximum was found in ceftazidime (78.4%), ofloxacin (76.1%) and ciprofloxacin (75.3%), the smallest in piperacillin/tazobactam (43.2%) and piperacillin (38.9%). The importance of incubation extended to 48 h during the testing of S. maltophilia strains was noted for correctly setting their susceptibility to antibiotics. PMID- 11061631 TI - Conservation in probiotic preparations of Lactobacillus with inhibitory capacity on other species. AB - Strains of Lactobacillus isolated from dairy products and genital tract competed with Candida albicans through a membrane of 12000 dalton cut-off. This inhibition was due to hydrogen peroxide and was trypsin-stable, heat-sensitive and antagonized by catalase. Lactobacillus coming from "starters" showed antimicrobial activity against fungus isolated in a yogurt factory. Penicillium, Alternaria, Phialophora, Microsporum and Candida spp. were inhibited when 10(2) spores were inoculated in the assay. No inhibition was observed with 10(5) spores. Besides, one of 21 Lactobacillus strains isolated from the vaginas of healthy women inhibited pathogenic bacteria by means a bacteriocin trypsin sensitive, heat-stable and retained by dialysis membrane. Tablets for future probiotic use were prepared and the viability of bacteria was assayed using media with different compositions. Pharmaceutical preparations with polyethyleneglycol was the best formulation for the Lactobacillus viability, the counts remained between 10(7) and 10(6) cfu/tablet for up to 1 year. PMID- 11061632 TI - Effect of sodium chloride and citric acid on growth and toxin production by A. caviae and A. sobria at moderate and low temperatures. AB - The effect of sodium chloride and citric acid on hemolysin and caseinase production by Aeromonas caviae and Aeromonas sobria at 32 degrees C and 5 degrees C was investigated. At 32 degrees C, although both strains were tolerant to 3% NaCl in TSB, the production of caseinase was decreased in the presence of 1-3% NaCl, and the production of hemolysin was abolished by 2-3% NaCl. Citric acid (0.03%) was less effective than NaCl in reducing hemolysin and caseinase production by both strains at 32 degrees C. A combination of low temperature (5 degrees C) and citric acid treatment reduced hemolysin and caseinase production by both strains. A combination of low temperature (5 degrees C) and NaCl (3%) treatment was the most effective procedure in reducing growth and hemolysin and caseinase production by the tested strains. PMID- 11061633 TI - Establishment of a canine monocyte cell line. AB - A canine monocyte cell line was established from the peripheral blood sample collected from a healthy young male Beagle dog. The cloned cells grew easily and were serially passaged in vitro in the medium, a slight modification further made on Dulbecco's modified Eagle medium, supplemented with 10% fetal bovine serum. The morphology of single cell was shown in triangular or round form, however, it became epithelioid in a densely grown monolayer. Non-specific esterase was detected in all cells by a cytochemical examination. The cells reacted rapidly to the addition of a small amount of LPS and differentiated to the cells of morphologically typical macrophages. Both complement receptor (CD35) and Fc gamma receptor (CD64) were demonstrated on the cell membrane. PMID- 11061634 TI - Absence of Helicobacter pylori in tonsillar swabs from dyspeptic patients. AB - Aim of this study was to evaluate whether tonsils might be a potential reservoir for Helicobacter pylori (H. pylori) infection. A total of 72 consecutive dyspeptic patients undergoing endoscopy for the first time were studied. For each patient, a bilateral tonsillar swab was performed, before endoscopy, for microbiological culture and immunochemical analysis. Antral biopsies were also collected at endoscopy for microbiological culture, rapid urease test, and histological examination. Helicobacter pylori infection was detected in 42 of 72 (58.3%) patients. All tonsillar specimens were negative for H. pylori on both microbiological culture and immunochemical analysis, suggesting that the tonsils are not an extragastric reservoir for H. pylori infection. PMID- 11061635 TI - Pathogenic mechanisms of Bartonella quintana. AB - Bartonella quintana is an epi- and intracellular gram-negative rod responsible for both acute and chronic clinical manifestations. We review the literature about pathogenic mechanisms of B. quintana and discuss our data. Our efforts to clarify Bartonella quintana pathogenesis run on two parallel tracks. The first one concerns interactions between Bartonella quintana and endothelial cells by evaluation and modulation of apoptosis, signal transduction pathways and inflammation. The second one concerns some biological activities of Bartonella quintana endotoxin on human whole blood and endothelium. The elucidation of the mechanisms regulating the inflammatory/proliferative pattern of chronic clinical manifestations of Bartonella quintana infections may offer a contribution for addressing the pathogenesis of intracellular bacterial persistence. PMID- 11061636 TI - Behavioral prevention is today's AIDS vaccine! AB - INTRODUCTION: Behavioral prevention is currently the only effective way to stem the further spread of HIV. This article reviews the pro-active programmatic model of behavioral research that has led to the development and testing of successful HIV/STD preventive interventions in the last fifteen years. OBJECTIVE: To present (1) a model of behavioral prevention adapting phases of clinical trials research: Phase I: Discovery; Phase II: Exploratory; Phase III: Efficacy; and Phase IV: Effectiveness; and (2) a theoretical framework for behavioral prevention; and (3) A Lifespan Model of Health Promotion and Disease Prevention which can be used to design HIV/STD prevention programs across the lifespan, at different levels (e.g., individual, couple, family, community, societal) using different intervention modalities. CONCLUSIONS: Behavioral prevention is effective with different age groups and at different levels of intervention when the prevention program has a theoretical basis. Behavioral prevention works now and can be mobilized within a community to address all of the factors associated with the rapid development of an epidemic. Behavioral prevention is cost effective and can be delivered in communities that have limited resources. PMID- 11061637 TI - An overview of the global HIV/AIDS epidemic, with a focus on the United States. AB - The HIV/AIDS epidemic is a global human tragedy, especially in sub-Saharan Africa. The pandemic affects people in the primer of their lives moving from at risk populations to broader cross-sections of society. There have been more than 47 million adults and children infected since the beginning of the epidemic, and more than 18.8 million people have died. Over 95% of the global total of all AIDS cases are in the developing world, with prevalence among adults at less than 1% in India and Europe, to more than 10% in several African countries. The overwhelming majority of all infections globally are acquired through unprotected sexual intercourse, with at least 70% resulting from heterosexual intercourse. There have been more than 733,374 AIDS cases reported to the Centers for Disease Control and Prevention (CDC) in the US since the beginning of the epidemic, and more than 430,000 deaths. The largest number and proportion of AIDS cases reported have occurred among gay and bisexual men. This trend continues today, although racial and ethnic minorities, women, and youth are becoming infected in increasing proportions. The south has the most people living with AIDS, followed by the north-east. The global situation is improving in some areas, but even if all HIV transmission could be completely stopped tomorrow, the long-term health, social and economic consequences will be devastating well into the 21st century. The magnitude of the epidemic and the continuing explosive risk of infection, coupled with the economic and infrastructural realities of the regions of the world, make prevention the only realistic approach. PMID- 11061638 TI - Multiple levels of analysis and intervention in HIV prevention science: exemplars and directions for new research. AB - The need to continue the scientific development of evidence-based HIV-1 prevention strategies is clear. The epidemic continues to rage out of control, and a vaccine against HIV-1 is nowhere in sight. We think that it is important to consider the influence of multiple social units on HIV risk behavior. In this article, we use ecological theory to outline multiple levels of analysis at which preventive interventions can be conceptualized. These levels include the individual, dyadic/small group, organizational, community, and societal/cultural. We discuss advantages and disadvantages of locating HIV risk at each level, and provide exemplars of HIV prevention for each in an effort to encourage HIV prevention scientists to consider the level at which they are locating the determinants of HIV risk behavior when conducting research. We conclude by stating that scientists and research funding has favored the individual level of analysis, but that to be most successful, the field of HIV prevention science should address risk behavior at all levels of analysis. PMID- 11061639 TI - Consequences of HIV prevention interventions and programs: spectrum, selection, and quality of outcome measures. AB - The outcome measures employed in an HIV prevention intervention study should match the research and policy questions at hand. If the question is 'did the intervention work to prevent HIV infection?', then seroincidence data may be insufficient. However, if the question is 'why did the intervention work?', then more detailed behavioral data are necessary (and sometimes behavior change itself is the real goal of an intervention study). Given the wide range of questions asked by HIV prevention policy makers, funders and researchers, a spectrum of outcome measures is needed across HIV prevention intervention studies. These include measures of behavioral determinants, HIV-related risk behaviors, HIV incidence (and other biologic markers), morbidity, mortality, and cost effectiveness factors (such as cost per quality-adjusted life year saved). In this paper, we review the range of outcome measures used and needed in these intervention studies. Particular attention is paid to the psychometric properties of self-reported behavior change measures of sexual behavior and substance use. Additional emphasis is placed on the role of cost-effectiveness measures in intervention studies. A general framework is proposed for conceptualizing the array of outcome measure possible for any given HIV prevention intervention study. PMID- 11061640 TI - HIV prevention interventions with gay or bisexual men and youth. AB - This article describes and reviews the findings of several well-designed and controlled outcome trials of HIV prevention interventions that have been undertaken with men who have sex with men. The interventions reviewed have been of two types: face-to-face group or workshop interventions and community-level programs undertaken in gay communities. Both have shown robust effectiveness in promoting risk reduction behavior change among gay men and young people at risk for contracting HIV infection. Conclusions can now be reached about the benefits of these types of interventions. However, there remains an urgent need to develop and tailor HIV prevention approaches that can promote the maintenance of behavior change; to reach community segments that remain vulnerable, especially young and minority men who have sex with men; and to address the changing context of the epidemic. PMID- 11061641 TI - HIV risk reduction behavioral interventions with heterosexual adolescents. AB - This article provides a comprehensive review of research on the effects of behavioral interventions on heterosexual adolescents' HIV sexual-risk behavior. It details adolescents' risk of sexually transmitted HIV infection and describes challenges associated with adolescent intervention research, including obtaining school and parent approval and the validity of self-reported measures. It describes central characteristics of 36 controlled intervention studies assessing the HIV sexual-risk behavior of over 30,000 male and female adolescents 11-21 years of age. It summarizes the participants' race/ethnicity and age, the theoretical framework, and the intervention setting, duration and outcome. This review reveals that the most commonly assessed behavioral outcomes were condom use and abstinence, and the largest effects sized were on condom use and condom acquisition. Effect sizes for abstinence and number of sexual partners were the smallest. Perceived self-efficacy and behavioral interventions were the most commonly assessed theoretical mediators. Key questions this research engaged in included whether behavioral skills can be increased, whether intervention-induced behavior change can be sustained, whether matching the race/ethnicity and gender of facilitators and participants enhances the effectiveness of culturally sensitive interventions, whether classroom teachers can effectively facilitate interventions, whether the behavior of high-risk populations can be changed, and which kinds of interventions are most effective. This review concludes that carefully designed theory-based interventions that take into account the characteristics of the particular population or culture can cause positive changes in adolescents' HIV sexual-risk behavior, but boundary conditions for their effectiveness still need to be identified. Several suggestions for additional research are proffered. PMID- 11061642 TI - Prevention of sexual risk behavior for HIV infection with women. AB - This paper defines the components of gender-specific interventions for HIV infections for women, i.e. negotiation skills with male partners for condom use, integration of strategies against HIV and other STD infections and for contraception, the urgent need for female controlled methods, the importance of the inclusion of heterosexual men and an expansion to couples in prevention programs. This paper also presents a critical update in HIV prevention articles for women since the beginning of the AIDS epidemic through March 1996. All reviewed interventions were conducted in the U.S., Canada or Puerto Rico and described a psychological, behavioral, or educational component that addressed sexual risk reduction and included a behavioral evaluation. Manual and computer searches identified 47 studies that targeted women and provided a female-specific analysis of intervention effects. Overall, the findings demonstrate that HIV prevention programs can be effective in reducing risky sexual behavior among women. Program effectiveness varied by intervention type, session duration, and whether studies included women alone or both men and women. The most efficacious HIV prevention programs were specifically directed toward women, focused on relationship and negotiation skills, and involved multiple, sustained contacts. Evidence also indicated that community-level interventions hold promise. It is recommended that outcomes for women be expanded to include strategies beyond the male condom, such as refusing or avoiding unsafe sex or using the female condom. PMID- 11061643 TI - HIV prevention programs with heterosexuals. AB - OBJECTIVE: Efficacious HIV prevention programs designed for heterosexual adults were identified. METHODS: Thirty-two programs designed with a comparison group and aimed at preventing heterosexual transmission for HIV were identified utilizing computerized data bases and key informants. RESULTS: Three types of efficacious interventions were identified: (1) those based on social cognitive theories that aimed to improve HIV-related knowledge, attitudes, norms, and behavioral practices (n = 27); (2) treatment of sexually transmitted diseases (STDs) (n = 3); and (3) pre- and post-test HIV testing and counseling programs (n = 2). The high incidence of HIV and STD in international settings has resulted in these trials demonstrating the greatest reductions in risk for HIV, reflected in biological markers of infection. Only five of 12 studies with injecting drug users were successful in reducing sexual risk behaviors. The optimal STD treatment strategy (syndromic case management, mass treatment) varies across communities. HIV testing and counseling appears an efficacious strategy, particularly for seropositive adults, yet current models have not considered the impact of new technologies on HIV testing paradigms. CONCLUSION: Each successful prevention strategy faces significant challenges before broad dissemination of the intervention approach can be achieved. PMID- 11061644 TI - Effectiveness of HIV prevention interventions in developing countries. AB - OBJECTIVE: To review the effectiveness of projects and programs in developing countries that aim to reduce sexual transmission of HIV infection or transmission related to injection drug use. DESIGN: We identified 34 published studies undertaken in 18 developing countries that met rigorous inclusion criteria. These criteria included the length of follow-up, use of statistical analysis, the inclusion of a comparison group, and type of outcomes measured. RESULTS: We found that behavioral change interventions are effective when targeted to populations at high risk, particularly female sex workers and their clients. Few studies have evaluated harm reduction interventions in injecting drug users (IDUs). Evidence on the effectiveness of voluntary counseling and testing programs was promising, and VCT was most effective when directed at discordant couples. Treatment of sexually transmitted diseases (STD) appears highly effective in reducing HIV/STD transmission, particularly in the earlier stages of the epidemic. CONCLUSIONS: This review demonstrates that HIV prevention interventions can be effective in changing risk behaviors and preventing transmission in low and middle-income countries. When the appropriate mix of interventions is applied, they can lead to significant reductions in the prevalence of HIV at the national level. Additional research is needed to identify effective interventions, particularly in men who have sex with men, youth, IDUs and HIV-infected persons. Structural and environmental interventions show great promise, although more evaluation is needed. PMID- 11061645 TI - Interventions to prevent HIV risk behaviors. National Institutes of Health Consensus Development Conference Statement February 11-13, 1997. AB - OBJECTIVE: To provide health care providers, patients, and the general public with a responsible assessment of behavioral intervention methods that may reduce the risk of HIV infection. PARTICIPANTS: A non-Federal, nonadvocate, 12-member panel representing the fields of psychiatry, psychology, behavioral and social science, social work, and epidemiology. In addition, 15 experts in psychiatry, psychology, behavioral and social science, social work, and epidemiology presented data to the panel and a conference audience of 1000. EVIDENCE: The literature was searched through Medline and an extensive bibliography of references was provided to the panel and the conference audience. Experts prepared abstracts with relevant citations from the literature. Scientific evidence was given precedence over clinical anecdotal experience. CONSENSUS PROCESS: The panel, answering predefined questions, developed its conclusions based on the scientific evidence presented in open forum and the scientific literature. The panel composed a draft statement that was read in its entirety and circulated to the experts and the audience for comment. Thereafter, the panel resolved conflicting recommendations and released a revised statement at the end of the conference. The panel finalized the revisions within a few weeks after the conference. CONCLUSIONS: Behavioral interventions to reduce risk for HIV/AIDS are effective and should be disseminated widely. Legislative restriction on needle exchange programs must be lifted because such legislation constitutes a major barrier to realizing the potential of a powerful approach and exposes millions of people to unnecessary risk. Legislative barriers that discourage effective programs aimed at youth must be eliminated. Although sexual abstinence is a desirable objective, programs must include instruction on safer sex behaviors. The erosion of funding for drug abuse treatment programs must be halted because research data clearly show that such programs reduce risky drug abuse behavior and often eliminate drug abuse itself. Finally, new research must focus on emerging risk groups such as young people, particularly those who are gay and who are members of ethnic minority groups, and women, in whom transmission of HIV virus to their children remains a major public health problem. PMID- 11061646 TI - Cidofovir added to HAART improves virological and clinical outcome in AIDS associated progressive multifocal leukoencephalopathy. AB - OBJECTIVES: To analyse the virological and clinical efficacy of cidofovir combined with highly active antiretroviral therapy (HAART) in AIDS-related progressive multifocal leukoencephalopathy (PML). DESIGN: Multicentre observational study of consecutive HIV-positive patients with histologically or virologically-proven PML. Group A, 26 patients treated with HAART; group B, 14 patients treated with HAART plus cidofovir 5 mg/kg intravenously per week for the first 2 weeks and alternate weeks thereafter. JC virus DNA was quantified in cerebrospinal fluid (CSF) by PCR. RESULTS: Baseline virological, immunological and clinical characteristics were homogeneous between the groups. In one case cidofovir was discontinued because of severe proteinuria. There was no significant difference in HIV RNA responses and changes in the number of CD4 cells between group A and B. After 2 months of therapy, five out of 12 (42%) patients from group A and seven out of eight (87%) from group B reached undetectable JC virus DNA in the CSF (Chi-square P = 0.04); moreover, 24% of group A and 57% of group B patients showed neurological improvement or stability (P = 0.038). One-year cumulative probability of survival was 0.67 with cidofovir and 0.31 without (log-rank test, P = 0.01). Variables independently associated with longer survival were the use of cidofovir, HAART prior to the onset of PML, a baseline JC virus DNA load in CSF < 4.7 log10 copies/ml, and a baseline Karnofsky performance status > or = 60. CONCLUSIONS: In AIDS-related PML, cidofovir added to HAART is associated with a more effective control of JCV replication, with improved neurological outcome and survival compared with HAART alone. PMID- 11061647 TI - Concurrent infections and HIV pathogenesis. PMID- 11061648 TI - Immune activation in africa is environmentally-driven and is associated with upregulation of CCR5. Italian-Ugandan AIDS Project. AB - BACKGROUND: HIV infection in Africa is associated with immune activation and a cytokine profile that stimulates CCR5 expression. We investigated whether this immune activation is environmentally driven; if a dominant expression of CCR5 could indeed be detected in African individuals; and if R5 HIV strains would be prevalent in this population. METHODS: Freshly drawn peripheral blood mononuclear cells from HIV-uninfected African and Italian individuals living in rural Africa, from HIV-uninfected Africans and Italians living in Italy, and from HIV-infected African and Italian patients were analysed. Determinations of HIV coreceptor specific mRNAs and immunophenotype analyses were performed in all samples. Virological analyses included virus isolation and characterization of plasma neutralizing activity. FINDINGS: Results showed that: immune activation is detected both in Italian and African HIV-uninfected individuals living in Africa but not in African subjects living in Italy; CCR5-specific mRNA is augmented and the surface expression of CCR5 is increased in African compared with Italian residents (CXCR4-specific mRNA is comparable); R5-HIV strains are isolated prevalently from lymphocytes of African HIV-infected patients; and plasma neutralizing activity in HIV-infected African patients is mostly specific for R5 strains. CONCLUSIONS: Immune activation in African residents is environmentally driven and not genetically predetermined. This immune activation results in a skewing of the CCR5 : CXCR4 ratio which is associated with a prevalent isolation of R5 viruses. These data suggest that the selection of the predominant virus strain within the population could be influenced by an immunologically driven pattern of HIV co receptor expression. PMID- 11061649 TI - Sperm washing and virus nucleic acid detection to reduce HIV and hepatitis C virus transmission in serodiscordant couples wishing to have children. AB - BACKGROUND: Use of a motile spermatozoa isolation process was assessed for reducing the transmission of HIV and hepatitis C virus (HCV) during artificial insemination in HIV-serodiscordant couples in which the man is infected. PATIENTS: Thirty-two HIV-1-infected clinically asymptomatic men, having a median CD4 cell count of 396 x 10(6)/l and a median blood plasma HIV-1 RNA content of 414 copies/ml. Of these, 16 were infected with both HIV and HCV. METHODS: Motile spermatozoa were isolated from 51 semen samples by density gradient and 'swim up'. HIV-1 and HCV genomes were detected and quantified in the blood plasma and seminal plasma, and detected in seminal cell fractions obtained during spermatozoa isolation. RESULTS: HIV-1 RNA was detected in 30% of seminal plasma samples. HIV-1 genomes were found in 18% of seminal cell samples, but in none of the motile spermatozoa fractions after 'swim-up'. There was no correlation between the HIV-1 RNA concentrations in the blood and seminal plasma. HIV-1 genome was detected intermittently in patients who gave more than one sample. HCV RNA was detected in 20% of seminal plasma samples from HCV viraemic patients, but in no seminal cells or motile spermatozoa fractions. CONCLUSIONS: Purification of motile spermatozoa by density gradient plus 'swim-up' reduced the HIV-1 and HCV genomes in the semen of infected individuals to undetectable levels. This method, associated with a standardized virus assay, could be useful for serodiscordant couples (males infected) who wish to have children. PMID- 11061650 TI - Effect of menstrual cycle on HIV-1 levels in the peripheral blood and genital tract. WHS 001 Study Team. AB - OBJECTIVE: To assess the variation in HIV-1 over the menstrual cycle, including RNA levels in the female genital tract, plasma HIV-1-RNA levels, CD4 cell counts, and culturable virus. DESIGN: A prospective analysis of 55 HIV-1-infected women. METHODS: Blood and genital tract specimens were collected weekly over 8 weeks, spanning two complete menstrual cycles. Applying repeated-measures models that used menses as the reference level, the variation in viral RNA levels was compared in endocervical canal fluid and cells (collected by Sno-strips and cytobrush, respectively) and ectocervicovaginal lavage (CVL) fluid. Repeated measures models were also used to assess the variation in plasma CD4 cell counts and viral load. RESULTS: Shedding patterns differed among the three sampling methods, independent of genital tract co-infections. Genital tract HIV-1-RNA levels from CVL fluid and endocervical canal cytobrush specimens were highest during menses and lowest immediately thereafter (P = 0.001 and P = 0.04). The HIV 1-RNA level in endocervical canal fluid was highest in the week preceding menses (P = 0.003). The menstrual cycle had no effect on blood levels of RNA (P = 0.62), culturable virus (P = 0.34), or CD4 cell counts (P = 0.55). HIV-1-RNA levels were higher in endocervical canal fluid than in peripheral blood plasma during the late luteal phase (P = 0.03). CONCLUSION: HIV-1-RNA levels vary with the menstrual cycle in the female genital tract but not the blood compartment. HIV-1 RNA levels are higher in endocervical canal fluid than in blood plasma. These findings may have important implications for sex-specific pathogenesis, heterosexual transmission, and contraceptive hormone interventions in HIV-1 infected women. PMID- 11061651 TI - Relationship of human herpesvirus 8 peripheral blood virus load and Kaposi's sarcoma clinical stage. AB - OBJECTIVE: To determine the relationship between human herpesvirus 8 (HHV-8 or Kaposi's sarcoma-associated herpesvirus) peripheral blood virus load and Kaposi's sarcoma (KS) clinical stage. DESIGN: Blinded, cross-sectional analysis of peripheral blood HHV-8 DNA levels in persons with AIDS-related KS in Harare, Zimbabwe. METHODS: Subjects were stratified by KS clinical stage. The amount of HHV-8 DNA in plasma and peripheral blood mononuclear cells (PBMC) was determined by quantitative real-time PCR amplification of the HHV-8 open reading frame 26. RESULTS: Thirty-one HIV-1/HHV-8-coinfected persons were studied: 26 subjects had histologically confirmed KS (one stage II, 11 stage III and 14 stage IV) and five subjects had antibodies to HHV-8 but did not have KS. The age, CD4 lymphocyte count and plasma HIV-1 RNA levels were similar in all groups. HHV-8 DNA was detected in the plasma of all HHV-8-infected subjects (range < 2.4 to 5.2 log10 copies/ml), but plasma HHV-8 DNA levels were not associated with KS disease stage. In contrast, the amount of HHV-8 DNA in PBMC (range < 0.7 to 4.5 log10 copies/microg) was strongly associated with KS clinical stage (P = 0.005). Among stage IV KS cases, there was a linear relationship between plasma and PBMC HHV-8 DNA levels (r2 = 0.42; P = 0.01). CONCLUSION: The strong association observed between the extent of KS disease and the levels of HHV-8 DNA in PBMC provides further evidence for a relationship between HHV-8 virus load and KS pathogenesis. PMID- 11061652 TI - Influence of CCR5 promoter haplotypes on AIDS progression in African-Americans. AB - OBJECTIVES: To test the hypothesis that the CCR5 promoter variants in HIV-1 infected African-Americans affect the rate of progression to AIDS and to determine the extent of linkage disequilibrium between the CCR5P1 allele and the CCR5 59029A variant (referred to here as CCR5-2459A), both of which have been shown independently to accelerate AIDS progression in Caucasians. DESIGN: We used survival analysis to assess the effects of CCR5 promoter variants in HIV-1 seroincident Caucasians and African-Americans. SUBJECTS AND METHODS: Genotypes were determined for 806 Caucasians and 1067 African-Americans, which included 700 seroconverters, enrolled in four HIV/AIDS natural history cohort studies. These genotypes were used to determine linkage and haplotypes for CCR2 and CCR5 alleles. Survival analysis was used to assess the effect of CCR2, CCR5, and CCR5 promoter haplotypes on progression to AIDS in seroincident African-Americans. RESULTS: A survey of Caucasians and African-Americans demonstrated complete linkage disequilibrium between CCR5P1 and CCR5-2459A sites. The composite CCR5P1 haplotype (including the CCR5-2459A allele) is shown to be associated with rapid progression to AIDS endpoints in both African-American and Caucasian cohorts, but the effect is recessive in Caucasians and dominant in African-Americans. This is probably due to the presence of modulating genes or as yet unidentified polymorphisms that may differ between racial groups. PMID- 11061653 TI - Clinical and metabolic presentation of the lipodystrophic syndrome in HIV infected children. AB - OBJECTIVE: To investigate body fat distribution and glucose and lipid metabolism in HIV-infected children with the aim of describing the lipodystrophic syndrome in children. DESIGN: Cross-sectional study including 39 HIV-infected children aged 3-18 years. MAIN OUTCOME MEASURES: Clinical lipodystrophy was defined as peripheral fat wasting (facial and/or buttock and/or limb atrophy with arm skinfold thickness lower than the third percentile of the reference values for sex and age) and/or truncal adiposity (breast enlargement and/or buffalo neck and/or relative abdominal obesity with trunk : arm skinfold ratio > 2 standard deviations). Fasting serum lipid concentrations were measured and an oral glucose tolerance test was performed. RESULTS: Of 39 HIV-infected children, lipodystrophy was observed in 13 children (33.3%): eight with truncal lipohypertrophy, three with peripheral lipoatrophy and two with combined lipodystrophy. Combined lipodystrophies were observed only in adolescents with a more severe presentation than in prepubertal children. Lipodystrophic children had higher fasting insulinaemia (7.0+/-8.5 versus 3.0+/-2.3 microU/ml; P = 0.07), suggesting a certain degree of insulin-resistance. Hypercholesterolaemia (23% versus 15%; P = 0.59 ) and hypertriglyceridaemia (15% versus 11%; P = 0.76) were observed with the same proportion in the lipodystrophic as in the non-lipodystrophic groups; 23% of the non-lipodystrophic children had dyslipidaemia. CONCLUSION: The lipodystrophic syndrome prevails in HIV-infected children in the three clinical forms initially described in adults but appears less severe before the initiation of puberty. Insulin-resistance occurs in lipodystrophic children only, whereas dyslipidaemia exists in non-lipodystrophic children as well, suggesting that dyslipidaemia could reflect subclinical alteration of the adipose tissue. PMID- 11061654 TI - The impact of protease inhibitor-containing highly active antiretroviral therapy on progression of HIV disease and its relationship to CD4 and viral load. AB - OBJECTIVE: To compare the rate of disease progression according to viral load and CD4 cell count in patients receiving or not receiving highly active antiretroviral therapy (HAART), defined as protease inhibitor-containing regimens. DESIGN: An observational study, with prospectively collected data. METHODS: All patients attending the HIV Outpatient clinic as of 1 January 1995 (n = 2083) were included. Follow-up was until the first AIDS-defining event or death. Associations between viral load or CD4 cell count and disease progression were assessed using a person-years approach. Event rates were compared using Poisson regression analysis; a multivariate model was used to assess the independent effects of CD4, viral load and treatment group on event rates and to consider interactions between these variables. RESULTS: The event rates increased with lower CD4 cell count and higher viral load for both treatment groups and were generally lower in the HAART group. In a multivariate analysis, lower CD4 cell counts and higher viral loads remained significantly associated with disease progression, irrespective of treatment group. However, the event rate was significantly lower for the HAART group compared with the control group (relative rate 0.53, P < 0.001). CONCLUSIONS: HAART-treated patients with high viral loads and CD4 cell counts experienced reduced disease progression compared with individuals with the same CD4 cell count and viral load not receiving HAART. Consequently, the short-term prognosis associated with viral load levels and CD4 cell counts may differ in patients on HAART. Whether this effect will be observed with non-protease-inhibitor-containing HAART is not known at this time. PMID- 11061655 TI - Zidovudine triphosphate and lamivudine triphosphate concentration-response relationships in HIV-infected persons. AB - OBJECTIVE: To quantitate intracellular concentrations of zidovudine and lamivudine triphosphate and explore relationships with virologic and immunologic responses to antiretroviral therapy. DESIGN: Eight antiretroviral-naive, HIV infected persons with CD4 T cell counts > 100 x 10(6) cells/l, and HIV RNA in plasma > 5000 copies/ml participating in a prospective, randomized, open-label study of standard dose versus concentration-controlled therapy with zidovudine, lamivudine, and indinavir. METHODS: Peripheral blood mononuclear cells and plasma were collected frequently throughout the study for quantitation of intracellular zidovudine triphosphate and lamivudine triphosphate concentrations, and zidovudine and lamivudine concentrations in plasma. CD4 T cells and HIV RNA in plasma (Roche Amplicor Ultrasensitive Assay) were measured at baseline and every 4 weeks throughout the study. Relationships among intracellular and plasma concentrations, and CD4 T cells and HIV RNA in plasma were investigated with regression analyses. RESULTS: Significant relationships were observed between the intracellular concentrations of zidovudine triphosphate and lamivudine triphosphate and the baseline level of CD4 cells. Lamivudine triphosphate concentrations were related in a linear manner to the apparent oral clearance of lamivudine from plasma. A direct linear relationship was found between the intracellular concentrations of zidovudine triphosphate and lamivudine triphosphate. The percent change in CD4 cells during therapy and the rate of decline in HIV RNA in plasma were related to the intracellular concentrations of zidovudine triphosphate and lamivudine triphosphate. CONCLUSION: These studies into the intracellular clinical pharmacology of nucleoside reverse transcriptase inhibitors illustrate potential clinical implications as determinants of therapeutic success. Moreover, these findings provide several leads and a strong impetus for future investigations with nucleoside reverse transcriptase inhibitors particularly when given in combination and sequentially. PMID- 11061656 TI - Long-term hydroxyurea in combination with didanosine and stavudine for the treatment of HIV-1 infection. Swiss HIV Cohort Study. AB - OBJECTIVE AND METHODS: In 1998 we reported on a randomized comparison between stavudine plus didanosine plus placebo versus stavudine plus didanosine plus hydroxyurea (HU), in patients with a CD4 count of 200-500 x 10(6) cells/l. After 3 months, the HU group had a higher proportion of patients with viral load < 200 x 10 cells/l. At the end of the 3 months blinded period, patients in the placebo group had the option to add HU if their viral load remained > 200 x 10(6) cells/l. We report results after 24 months. RESULTS: Seventy-two patients were randomized to the HU arm, and a further 30 elected to add HU after 12 weeks. Twenty-four months after the start of the trial, only 25% of the 72 patients originally randomized to HU, and 20% of the 30 who added HU after week 12, were still taking it. The reasons for stopping HU were: lack of efficacy (45%), adverse events (37%) and patient or physician preference (18%). Side effects were more frequent in the didanosine/stavudine/HU group than in the didanosine/stavudine group: neuropathy (35 versus 15%, P< 0.02), fatigue (22 versus 7%, P< 0.01), and nausea or vomiting (26 versus 9%, P< 0.01). Of those who had discontinued HU, 73% were taking three drugs including a protease inhibitor. Patients who had started HU were compared with similar patients who had started protease inhibitors in the Swiss cohort. The probability of stopping HU was higher than the probability of stopping nelfinavir or indinavir, and similar to the probability of stopping ritonavir. CONCLUSION: HU increased the antiviral effect of stavudine plus didanosine. However, side effects were more frequent, and after 24 months the majority of patients had switched to protease inhibitor regimens. PMID- 11061657 TI - Prevention of nevirapine-associated exanthema using slow dose escalation and/or corticosteroids. AB - OBJECTIVE: The appearance of rash is one of the most frequent and limiting side effects during the first 4 weeks of treatment with nevirapine (NVP). We explored the efficacy and safety of four different strategies for reducing the incidence of this complication. PATIENTS AND METHODS: Four-hundred and sixty-nine patients were assigned randomly to accomplish the induction phase of NVP following either the standard recommendation of 200 mg daily during the first 2 weeks (n = 166), or any of three new strategies: adding prednisone 50 mg each other day during the first 2 weeks (n = 93); using a slowly escalating dose, beginning with 100 mg daily the first week, and increasing the dose by 100 mg/week up to the full daily dose of 400 mg (n = 107); and combining both the addition of prednisone with the slowly escalating dose (n = 103). A pharmacokinetic substudy was performed in seven patients receiving 100 mg of NVP during the first week. RESULTS: The incidence of rash diminished from 18.7% using the standard recommendation to 9.2% using the alternative approaches (P = 0.003). Rash appeared in 11.2%, 8.6%, and 7.7% of subjects assigned to receive the slowly escalating dose, prednisone, or both, respectively, without significant differences among them. The rate of drug discontinuation was also diminished by one-half using the new approaches (8.5% versus 4.3%; P = 0.06). NVP plasma concentrations within the first week of treatment using 100 mg daily were above the 90% inhibitory concentration for wild type HIV-1 in all instances. CONCLUSION: The incidence of rash complicating the first few weeks of treatment with NVP can be diminished by adding corticosteroids for 2 weeks to the standard recommendation, or by using a slowly escalating dose. This second approach is proven to be pharmacokinetically safe. PMID- 11061658 TI - The cost-effectiveness of expanded HIV counselling and testing in primary care settings: a first look. AB - OBJECTIVE: To estimate the cost-effectiveness of approaches to expanded HIV counselling and testing. DESIGN: A cost-effectiveness analysis. SETTING: Primary care practices in the USA. PARTICIPANTS: New patient visits. INTERVENTIONS: Two approaches were examined: (i) requesting all patients to complete an HIV-risk screening instrument, with counselling as well as testing offered only to patients disclosing risk factors ('risk histories' option); and (ii) routine offering of voluntary testing to all patients, with consent obtained but no pre test counselling ('routine testing'). MAIN OUTCOME MEASURES: The primary outcome was the cost per infection identified. We also examined: (i) the costs and numbers of infections averted if individuals change their risk behaviours; and (ii) the additional years of life and quality-adjusted life years (QALY) gained as a result of earlier HIV testing and treatment for infected individuals. RESULTS: Routine testing is the most cost-effective approach to identifying infected individuals at an incremental cost of US$4200 per infection identified. Although using risk histories is more costly and less effective than routine testing, it becomes similarly cost-effective using plausible ranges for sensitivity analyses. If at least 10% of HIV-positive individuals change their behavior, both routine testing and using risk histories would save money. If testing identifies infected individuals one year earlier than they otherwise would have been diagnosed, routine testing would cost US$22000 per QALY gained. CONCLUSION: Routine testing is the most cost-effective approach to identifying new HIV infections. However, using risk histories may be similarly cost-effective under various assumptions. Both routine testing and using risk histories are more cost-effective than current practices. PMID- 11061659 TI - Pregnancies before and after HIV diagnosis in a european cohort of HIV-infected women. European Study on the Natural History of HIV Infection in Women. AB - OBJECTIVES: Because most HIV-infected women are of reproductive age, we investigated whether their reproduction planning was affected by their HIV diagnosis. DESIGN: The European women study is a prospective, multicentre cohort of 485 HIV-infected women with a known interval of seroconversion. METHODS: The incidence of pregnancy was measured with person-time methods. Generalized estimating equation analysis was used to determine risk factors for pregnancy and pregnancy outcomes. RESULTS: In 449 women, the age-adjusted incidence of pregnancies decreased from 8.6 before HIV diagnosis to 8.2 and 6.0 per 100 person years in 0-4 and over 4 years after HIV diagnosis, respectively (P = 0.14). The proportion of induced abortions increased from 42% before to 53% after HIV diagnosis (P < 0.05). The risk of spontaneous abortion did not increase as a result of HIV infection. Since 1995, the proportion of births increased (P = 0.009), whereas that of induced abortions decreased (P = 0.01) compared with earlier years. An increased risk of pregnancy after HIV diagnosis was found in northern and central European women compared with southern European women; there was a lower risk in single women than in women with a steady partner. Of all pregnant women, single women, women between 15 and 25 years of age, and women with multiple partners were at increased risk for induced abortion both before and after HIV diagnosis. CONCLUSION: The incidence of pregnancy decreased with HIV disease progression. Pregnancies after HIV diagnosis appear to be related largely to social and cultural attitudes. The number of induced abortions was high before HIV diagnosis and it significantly increases thereafter. PMID- 11061660 TI - Back-calculation by birth cohort, incorporating age- specific disease progression, pre-AIDS mortality and change in European AIDS case definition. European Union Concerted Action on Multinational AIDS Scenarios. AB - OBJECTIVES: To adapt and improve methodology for back-calculation of AIDS in Europe and to examine the feasibility of estimating past HIV incidence by birth cohort. METHODS: Empirical Bayesian back-calculation (EBBC) used Markov disease progression models, modified to allow for three diseases added to the AIDS case definition in 1993 and for pre-AIDS mortality, and estimation by penalized maximum likelihood with a neighbour prior. EBBC by 5-year birth cohort assumed a minimum age at infection and age-dependent progression rates; three versions, with varying age effects, were investigated using AIDS cases diagnosed prior to the introduction of highly active antiretroviral therapies (HAART). RESULTS: Compared with the no age-effect version, EBBC by birth cohort tended to produce flattened HIV incidence curves in country-exposure groups with < 1000 AIDS cases, reflecting effects of the neighbour prior when data become sparse. Otherwise, birth cohort analysis, with moderate effects of age on progression, gave initially increasing incidence curves and consistent patterns across countries, with the 1960-1964 cohort most affected. In the European Union, incidence is estimated to have peaked in 1983 among homosexual men and in 1988 among injecting drug users; 460000 persons were estimated to be living with HIV/AIDS at the end of 1995. CONCLUSIONS: Our improved methodology deals effectively with the change in AIDS case definition and has allowed quantitative assessments of the HIV epidemic by birth cohort using all AIDS cases diagnosed before 1996, thus providing a sound basis for public health policy at a time when estimation of more recent prevalence is compromised by the effects of HAART. PMID- 11061661 TI - The Atlanta Urban Networks Study: a blueprint for endemic transmission. AB - OBJECTIVE: To study prospectively social networks and behavior in a group of persons at risk for HIV because of their drug-using and sexual practices, with particular emphasis on the interaction of risks and concomitant network structure. METHODS: A longitudinal study was conducted of 228 respondents in Atlanta, Georgia in six inner-city community chains of connected persons, interviewing primary respondents and a sample of their contacts every 6 months for 2 years. Ascertained were: HIV and immunologic status; demographic, medical, and behavioral factors; and the composition of the social, sexual, and drug-using networks. RESULTS: The prevalence of HIV in this group was 13.3% and the incidence density was 1.8% per year. Substantial simultaneity of risk-taking was observed, with a high level of both non-injecting (crack, 82%) and injecting (heroin, cocaine or both, 16 30%) drug use, the exchange of sex or money for drugs by men (approximately 35%) and women (57-71%), and high frequency of same sex sexual activity by men (9.4%) and women (33%). The intensity of interaction, as measured by network features such as microstructures and concurrency, was significantly greater than that observed in a low prevalence area with little endemic transmission. CONCLUSION: The traditional hierarchical classification of risk for HIV may impede our understanding of transmission dynamics, which, in the setting of an inner-city population, is characterized by simultaneity of risk taking, and moderately intense network interactions. The study provides further evidence for the relationship of network structure to transmission dynamics, but highlights the difficulties of using network information for prediction of individual seroconversion. PMID- 11061662 TI - Novel HIV-1 co-receptor-CCR5 promoter mutations in simians: identification of two highly polymorphic regions with extensive deletions. PMID- 11061663 TI - Drug-related behaviour in a high HIV prevalence rate population at Myktyina drug treatment centre, Kachin State, northern Myanmar (Burma). PMID- 11061664 TI - Detection of genotypically drug-resistant HIV-1 variants and non-B subtypes in recently infected antiretroviral-naive adults in Italy. PMID- 11061665 TI - Expression of sarcolectin in sera of HIV-1-infected patients during progression of the disease. PMID- 11061666 TI - Counselling to pregnant HIV-seropositive women with regard to feeding their babies. PMID- 11061667 TI - Pharmacokinetics of indinavir and low-dose ritonavir in children with HIV-1 infection. PMID- 11061668 TI - Circulating levels of IL-18 in adult and paediatric patients with HIV-1 infection. PMID- 11061669 TI - IL-18 underexpression reduces IL-2 levels during HIV infection: a critical step towards the faulty cell-mediated immunity? PMID- 11061670 TI - Osteonecrosis of the femoral head in HIV-1 patients: four additional cases. PMID- 11061671 TI - Patterns of lymphotropic herpesvirus viraemia in HIV-infected patients with Kaposi's sarcoma treated with highly active antiretroviral therapy and liposomal daunorubicin. PMID- 11061672 TI - Highly active antiretroviral therapy does not protect against Kaposi's sarcoma in HIV-infected individuals. PMID- 11061673 TI - Bone mineral loss in HIV-positive patients receiving antiretroviral therapy. PMID- 11061674 TI - HIV risk behavior among bisexual and heterosexual drug users. AB - This study examined the sexual and drug use behaviors for bisexual and heterosexual drug users (n=11,435 males and n=5,636 females) who participated in the NIDA AIDS Cooperative Agreement study. Results of the study suggest that, for males, bisexuality was highly associated with being homeless, having ever been paid for sex, having five or more sex partners in the month preceding the interview, having an IV drug-using sexual partner in the month preceding the interview, using crack, and sharing injection equipment in the month preceding the interview. For females, bisexuality was associated with ever having been arrested, past substance abuse treatment, ever having been paid for sex, ever having paid for sex, having five or more sexual partners in the month preceding the interview, ever using cocaine, and sharing injection equipment in the month preceding the interview. Overall, results from this study indicate that both male and female bisexuals, when compared to heterosexuals, were at higher risk for HIV and were more likely to be HIV positive. One implication of these results is that a universal prevention message may not be as effective as targeting prevention messages specifically for bisexual males and females. PMID- 11061675 TI - Drug information libraries on the Internet. AB - In the past several years there has been a dramatic proliferation of drug-related sites on the Internet. This article reviews the information found at selected Internet drug information libraries, and comments on its accuracy and implications. Drug-related sites were found by initially performing an Internet search on "psychoactive drugs" and then exploring links among the sites identified. Sites were chosen on the basis of comprehensiveness of information and positive or tolerant attitude toward drug use. While all classes of drugs are discussed at these sites, the primary foci of interest are synthetic and naturally occurring hallucinogens. Many of the biological materials discussed are legal and readily available. Information surveyed at these sites was largely accurate regarding the effects of various substances and biological sources of psychoactive compounds. CONCLUSIONS: Internet drug information libraries contain large amounts of information about a wide variety of drugs, including previously little-known biological sources of hallucinogens. The availability of this information could have significant effects on patterns of drug use. PMID- 11061676 TI - Sex trading and psychological distress in a street-based sample of low-income urban men. AB - This article examines the relationship between sex trading and psychological distress and assesses sexual human immunodeficiency virus (HIV) risk behaviors and HIV seroprevalence in a sample of young men recruited from the streets of Harlem. The authors interviewed 477 men, aged 18 to 29 years, of whom 43 (9.0%) had received money or drugs in exchange for sex in the preceding 30 days and were categorized as sex traders. Psychological distress was measured by using the Brief Symptom Inventory (BSI). Sex traders scored significantly higher than non sex traders on the General Severity Index and on all nine subscales of the BSI. According to multivariate analysis after adjusting for perceived HIV risk, current regular crack cocaine use and homelessness, sex traders scored 0.173 units higher on the General Severity Index than non-sex traders (p < .001). More of the sex traders tested positive for HIV (41% versus 19%, p < .001). The alarmingly high HIV seroprevalence rate in sex traders in this sample underscores the need to redouble HIV prevention efforts for this population. The high levels of psychological distress and crack cocaine dependence among sex traders may undermine their ability to adopt safer sex behaviors and should be considered in intervention designs. PMID- 11061677 TI - Blood alcohol content and death from fatal injury: a study in the metropolitan area of Sao Paulo, Brazil. AB - This study analyzed 5,690 toxicological screenings carried out on blood and viscera of fatally injured victims at the Institute of Forensic Medicine in the Metropolitan area of Sao Paulo during 1994. The screenings analyzed correspond to 39.5% of all deaths due to injury in this metropolitan area during the same period. Almost half of the victims (48.3%) presented a positive blood alcohol content (BAC). The exact proportion however, varied according to the cause of death with 64.1% of victims of drowning testing positive for alcohol, 52.3% of homicides, 32.2% of suicides and 50.6% of motor vehicle accidents. Blood alcohol concentration was also found to vary, with suicide victims presenting low concentrations and 70% of pedestrians hit by cars presenting high concentrations (0.2% or greater). Few cases tested positive for drugs other than alcohol, and of those who did, the majority were positive for cocaine. However, it should be emphasized that the methods used for the detection of substances other than alcohol were only accurate enough to detect cases of overdose. These findings highlight the need to improve surveillance of alcohol-related fatalities in Brazil and suggest an important link between alcohol intoxication and fatal injury. PMID- 11061678 TI - A comparative clinical study of the effect of WeiniCom, a Chinese herbal compound, on alleviation of withdrawal symptoms and craving for heroin in detoxification treatment. AB - WeiniCom is a Chinese herbal compound. The purposes of this double blind study were to evaluate (1) the efficacy of WeiniCom in reducing acute opioid withdrawal symptoms and craving, and (2) the side effects of WeiniCom, in each instance by comparing WeiniCom with buprenorphine, an established opioid detoxification treatment agent. Forty-two heroin addicts meeting the criteria of dependence in DSM-IV were randomly assigned to two treatment groups: a WeiniCom group (21 cases), and a buprenorphine group (21 cases). The Withdrawal Symptom Rating Scale and the Craving Rating Scale were employed to assess acute withdrawal symptoms and craving for heroin, and the Side Effects Rating Scale was used to measure side effects in the 14-treatment period. Both the WeiniCom and buprenorphine treatments are well-tolerated and very safe. Overall, the relief from opioid withdrawal symptoms and craving was better in the WeiniCom group than in the buprenorphine group. The rate of reduction in the severity of the withdrawal symptoms was faster in the WeiniCom group than in the buprenorphine group. By day nine to 10, the WeiniCom group showed very few withdrawal symptoms. In contrast, from day five on, the buprenorphine group continued to report relatively high scores for withdrawal symptoms and craving. WeiniCom demonstrated positive effects quickly, and required a shorter treatment period to achieve a desired degree of elimination of acute withdrawal symptoms and craving. PMID- 11061679 TI - Adoption of injection practices in heroin users in Guangxi Province, China. AB - This preliminary study collected data from 326 heroin users in Guangxi Province, China, in 1997. Logistic regression analysis was performed to identify the risk factors for injection. Survival analysis identified factors independently associated with time from initiation of heroin use to adoption of injection. Four factors were independently associated with injection: number of friends who used heroin in the last year, duration of heroin use, dose of heroin consumed, and total number of times detoxified in drug treatment and rehabilitation centers. Only gender and duration of heroin use were independently associated with time to first injection. Median time to first injection was 11 months for males and 22 months for females. Median time to first injection varied by age. Median time to injection for those who used heroin for more than one year was 8.1 months; it was 19.1 months for on to five years of use, and 40.5 months for more than five years of use. This study's preliminary findings suggest that younger, more recent heroin users, and males are at increased risk of becoming injectors, a major risk behavior for HIV acquisition. PMID- 11061680 TI - Prevalence and treatment of substance abuse in the mentally retarded population: an empirical review. AB - This article presents the first comprehensive review of studies of alcohol and illicit substance use in mentally retarded individuals, including prevalence, and recommendations for assessment and treatment. Mentally retarded persons appear to use/abuse alcohol at about the same rate as their noncognitively-impaired counterparts, and illicit drugs at moderately lower rates. However, little is known regarding which assessments and interventions are most effective in this population, given the absence of published treatment outcome studies and case examples. This is particularly disconcerting as detrimental consequences resulting from substance use have been identified in mentally retarded samples. Anecdotal data suggests that treatment for these individuals require modifications of existing empirically-derived substance abuse interventions to accommodate their unique needs. PMID- 11061681 TI - Clinical judgments of high-risk behavior during recovery. AB - This study utilized focus group research to explore high-risk behavior during recovery from drugs and alcohol. Participants in the focus group were professional substance abuse counselors. The findings identified specific high risk behaviors and began an exploration of the processes that support them. Specific information is included on such issues as causes, time frames, developmental issues, and other factors associated with high-risk behavior. Attention was paid to existential issues in recovery, as well as childhood factors such as sexual abuse. High-risk behavior was seen as a means to avoid the existential dilemma of continuing in recovery or returning to drug use and as a possible means to leave this crisis unresolved without actual relapse to drug use, or dealing with the issues of advancing in recovery. PMID- 11061682 TI - Cessation of drug use: impact of time in treatment. AB - Many studies have found that the longer a drug user remains in treatment, the more positive the outcome. The majority of studies on the effects of time in treatment have followed subjects from the time they enter treatment. The subjects of the present study are injection drug users and crack users who were out of treatment at the time of their recruitment to the study. Between the initial and six-month follow-up interviews, some chose to enroll in drug treatment. The more time a subject spent in treatment during the follow-up period, the more likely it was that s/he was not using heroin or cocaine at follow-up (OR=.51; 95% C.I., .39 .67; p<.001). Unlike the results of some prior studies, positive effects of time in treatment were found even when time in treatment was less than 90 days. The findings of the present study strongly suggest that treatment is beneficial even for those who remain for less than 90 days. Those who provide treatment services to drug users should attempt to maintain contact with dropouts, and support their return to treatment. PMID- 11061683 TI - Contingency contracting and systematic desensitization for heroin addicts in methadone maintenance programs. AB - The use and effectiveness of contingency contracting and systematic desensitization with heroin addicts being treated in methadone maintenance programs are discussed. Both behavior therapies can be practically implemented in methadone maintenance programs to supplement methadone pharmacotherapy. Contingency contracting has been effectively employed to reduce illicit drug use and to manage patients in the clinic. Systematic desensitization has less effect on actual heroin usage yet effectively reduces the fear of withdrawal and general anxiety, while improving self-image, assertiveness, and adjustment in the community. A clinic protocol that incorporates all three therapies-methadone maintenance, contingency contracting, and systematic desensitization-is proposed. PMID- 11061684 TI - Bufotenine: toward an understanding of possible psychoactive mechanisms. AB - A review of the neuropharmacology of the alleged hallucinogen bufotenine is presented, including recent experimental results showing activity similar to LSD and other known hallucinogens (psilocin and 5-MeO-DMT) at the purported hallucinogenic serotonin (5-HT) receptors, 5-HT2A and 5-HT2C. In addition, current reports of computer modeling of the receptors and ligand binding sites give evidence of bufotenine's ability to bind and activate these receptors. While binding and activation of the purported hallucinogenic receptors are not the full extent of the hallucinogenic signature, this evidence shows support for the rationale that the reported lack of the drug's classic hallucinogenic response in human experiments is due to poor ability to cross the blood brain barrier (BBB), not lack of activation of the appropriate brain receptors. Further evidence is reviewed that in some physiological states, some drugs with characteristics similar to bufotenine which do not normally cross the BBB, cross it and enter the brain. While direct human experimental evidence of bufotenine's hallucinogenic activity seems lacking, the above combined factors are considered, and possible explanations of bufotenine's reported psychoactivity are suggested. Additionally, updated experimental models testing the possible nature of bufotenine's hallucinogenic potential are proposed. PMID- 11061685 TI - After the drinking stops: completed suicide in individuals with remitted alcohol use disorders. AB - A substantial minority of suicide victims have remitted alcohol use disorders. The authors hypothesized that psychiatric disorders are likely to be present in this group to create the necessary conditions for suicide. They compared suicide victims with active alcohol use disorders and those with remitted alcohol use disorders. Using data on a community sample of suicide victims (N=141), it was determined that 39% (N=55) had a history of alcohol misuse. These subjects were categorized by alcohol use disorder status (remitted versus active) and by age (<50, > or =50), creating four cells: younger remitted (N=8), older remitted (N=9), younger active (N=22), and older active (N=16) alcohol misusers. Results comparing proportions of DSM-III-R Axis I disorders in the four groups indicated that suicide victims with remitted alcohol use disorders were predominantly younger victims with psychotic disorders or older victims with major depression. These findings have implications for identifying those at risk for suicide even after the cessation of alcohol misuse. Case examples of suicide victims with remitted alcohol use disorders-a younger woman victim with a psychotic disorder and an older man with major depression-are presented. PMID- 11061686 TI - Issues of fatherhood and recovery for VA substance abuse patients. AB - Drug-addicted fathers bring to treatment many uncertainties about their relevance to their children. Whether they are in contact with their children or not, they often believe their children are better off without contact with them. In working with these fathers, the authors have observed these men raising a number of issues concerning the father role. These include having no concept of what a father should be, confusing the roles of manhood and fatherhood, feeling inadequate as a provider, and not knowing how to reconnect with children they have not seen, particularly daughters. The fathers also have to learn to deal with their own guilt concerning their abandonment of their children. Suggestions for interventions with the fathers are given and include offering a workshop for fathers where they are shown visual images of positive fathering and can discuss their own parenting experiences. PMID- 11061687 TI - The poet syndrome: opiates, psychosis and creativity. AB - For some people, heroin is a self-medicating tool used to control innate psychic sensitivity. The habitual use of heroin provides the sensitive addict with a definition of consciousness by containment, psychic buffering, and psychic marking. This article is an examination of the political and social history of opiates, opiates as antipsychotics, "drug of choice" as a determinant of self medication, and the connection between creativity, spirituality, psychosis and addiction. Using clinical observation, the article explores the poet syndrome hypothesis and offers direction for an alternative drug treatment paradigm. PMID- 11061688 TI - Bone tumor dynamics: an orthopedic pathology perspective. AB - The diagnosis and classification of primary bone tumors remains as much a challenge today as it has for the last 80 plus years. Although pathology is invariably equated with the image of a diagnostic microscope, the vast majority of diagnoses are made grossly with the unaided eye, as are the tissue specimens selected for microscopic "confirmation." Radiologic studies, particularly plain radiographs, remain the gold standard in gross pathologic diagnosis of the skeleton. Today, confirmation and final classification continue as the pathologist's domain, but perhaps not for long, considering the evolving ancillary imaging techniques and progressive sophistication of magnetic resonance (MR) imaging. The bone tumor cases collected and compiled by Ernest Codman, M.D. during the second through fourth decades of this century formed the basis of the first tumor registry. The Codman Bone Sarcoma Registry demonstrated among other things the importance of radiographic/pathologic correlation, underscoring the reliability of a bone tumor's location, margin (host bone/tumor interface), periosteal reaction, and matrix patterns as an accurate guide to classification and likely future biologic behavior. "A General Theory of Bone Tumors," written by Lent C. Johnson nearly 50 years ago and published in the Bulletin of The New York Academy of Medicine (February 1953, second series, vol. 29, no. 2, pp. 164 171), provided a conceptual cellular approach to the understanding bone tumor dynamics reinforcing radiologic/pathologic correlation as a reliable diagnostic tool. At the time of Dr. Lent C. Johnson's death (1910-1998), he was literally working on an updated version of his original article, the latter of which is being reprinted as the core of this illustrated revision. Our continued experience with bone tumors over the past five decades has only served to validate, on a daily basis, the fundamental principles outlined in Johnson's original article. In like fashion, it is important to keep in mind that terminology and nomenclature has also evolved since 1953, despite a continued inability to achieve complete consensus. PMID- 11061689 TI - Staging of bone neoplasms: an orthopedic oncologist's perspective. AB - The process of staging bone tumors is complex. The goal of staging is to define the type of tumor and its extent. Like staging for other neoplasms, it stratifies patients into groups based on prognosis and established treatment protocols. Staging is a multidisciplinary effort involving orthopedic oncologists, musculoskeletal radiologists, and orthopedic pathologists. The diagnosis is often suggested on clinical examination and review of the radiographs. The biopsy usually confirms the clinical and radiographic impression. However, biopsy is difficult and leads to errors in diagnosis in nearly 20% of cases. These errors may make limb salvage impossible and adversely affect survival. For this reason, staging and especially the biopsy should be done in the institution where definitive treatment is planned. PMID- 11061690 TI - Osteoid-producing tumors of bone. AB - Osteoid-producing skeletal tumors are commonly encountered lesions by radiologists. Lesions include both benign tumors--osteoma, enostosis, osteoid osteoma, and osteoblastoma--as well as the malignant neoplasm, osteosarcoma. Radiologic features of these lesions are emphasized in this article and are frequently diagnostic. In addition, clinical characteristics, treatment, and prognosis are reviewed. PMID- 11061691 TI - Benign chondroid neoplasms of bone. AB - Benign cartilage lesions discussed in this article include osteochondroma (solitary, epiphyseal, and multiple), chondroblastoma, periosteal chondroma, and chondromyxoid fibroma. These lesions often demonstrate imaging appearances strongly suggesting the above diagnosis, particularly the "ring and arc" mineralization characteristic of cartilage lesions, which reflects their underlying pathology. This article emphasizes the imaging spectrum of these lesions with a multimodality approach. PMID- 11061692 TI - Enchondroma and chondrosarcoma. AB - Enchondroma and chondrosarcoma are two of the most commonly encountered primary bone lesions in the typical radiology practice. The purpose of this article is to review the clinical, radiological, and pathological features that distinguish conventional chondrosarcoma from enchondroma. Chondrosarcoma is almost always associated with pain and tends to present in the axial skeleton of middle aged adults. Enchondroma tends to present in young adults in the appendicular skeleton, particularly the hands, and is often an incidental finding. Although both lesions have characteristic radiographic appearances, difficulty separating these two entities most often occurs when a lesion is seen in the long tubular bones. The judicious use of computed tomography, magnetic resonance imaging, and nuclear medicine in conjunction with appropriate clinical data allows the radiologist to establish the correct diagnosis of benign or malignant medullary chondroid lesion in the majority of cases. PMID- 11061693 TI - Primary musculoskeletal tumors of fibrous origin. AB - Tumors of fibrous origin include fibrous dysplasia (FD), fibroxanthoma (nonossifying fibroma), cortical desmoid, desmoplastic fibroma, fibrosarcoma, and malignant fibrous histiocytosis (MFH). Benign fibrous lesions (FD, fibroxanthoma, and cortical desmoid) frequently demonstrate pathognomonic radiologic characteristics obviating the need for biopsy. Indeed, biopsy of these lesions can occasionally lead to confusion with more aggressive lesions. Desmoplastic fibroma and the malignant fibrous lesions (fibrosarcoma and MFH) often reveal nonspecific imaging features of a solitary nonmineralized lesion with aggressive characteristics. However, imaging is important as with other neoplasms in delineating the extent of involvement for staging purposes. This article reviews the spectrum of clinical characteristics, pathology, imaging appearances, treatment, and prognosis of lesions of fibrous origin in bone. PMID- 11061694 TI - Alphabet soup: cystic lesions of bone. AB - Giant cell tumor (GCT), aneurysmal bone cyst (ABC), and simple bone cyst (SBC) represent relatively common tumors and tumorlike conditions to affect bone. This article describes the clinical presentation of these lesions, as well as the characteristic radiologic and pathologic findings of each. In addition, differential diagnoses, disease course, and various treatment options are discussed. PMID- 11061695 TI - Angiomatous skeletal lesions. AB - Vascular lesions involving osseous structures are relatively common neoplasms. We will review the appearance of many musculoskeletal angiomatous lesions using the multimodality approach. Lesions to be discussed include osseous hemangioma, glomus tumor, angiomatosis and associated syndromes (Osler-Weber-Rendu, Klippel Trenaunay-Weber, and Kasabach-Merritt), Gorham, tumor-induced osteomalacia, and aggressive vascular neoplasms (hemangioendothelioma, hemangiopericytoma, and angiosarcoma). PMID- 11061696 TI - Lesions of unknown histogenesis: Langerhans cell histiocytosis and Ewing sarcoma. AB - Langerhans cell histiocytosis (LCH) and Ewing sarcoma represent lesions of unknown histogenesis. Both lesions typically affect children and adolescents. Imaging features frequently suggest the diagnosis, and both lesions may demonstrate aggressive characteristics. LCH shows a broader spectrum of imaging findings, depending on lesion activity and location, which are reviewed in this article. Ewing sarcoma typically reveals both osseous and soft tissue involvement, although direct continuity of the components is often not apparent. Knowledge of the spectrum of clinical and radiologic features facilitates early diagnosis and can be used to evaluate the effects of therapy. PMID- 11061697 TI - Myeloma and lymphoma. AB - This article reviews current information regarding myeloma, especially multiple myeloma and plasmacytoma as well as primary lymphoma of bone (PLB). The emphasis is on diagnostic imaging, including radiography, computed tomography (CT), magnetic resonance (MR) imaging, and skeletal scintigraphy. Relevant clinical information, histologic findings, treatment, and outcome data are also discussed. PMID- 11061698 TI - Imaging evaluation of the response of high-grade osteosarcoma and Ewing sarcoma to chemotherapy with emphasis on dynamic contrast-enhanced magnetic resonance imaging. AB - Response of high-grade osteosarcoma and Ewing sarcoma to preoperative chemotherapy can significantly affect the surgical approach to patients with these tumors and their disease-free survival. This article presents the findings and limitations of different imaging modalities (radiography, angiography, sonography, scintigraphy, and magnetic resonance [MR] imaging and spectroscopy) for evaluating this chemotherapeutic response. Particular emphasis is given to the currently most effective modality--dynamic contrast-enhanced MR imaging. PMID- 11061699 TI - Shoulder and humerus trauma. AB - Three bones and two joints comprise the pectoral girdle. These are the clavicle, scapula, proximal humerus, acromioclavicular joint, and shoulder joint. The pectoral girdle binds the upper extremity to the torso. Thus, injury to any of the pectoral girdle's components can impact the function of the entire extremity. The spectrum of pectoral girdle injuries ranges from a simple acromioclavicular joint sprain to scapulothoracic dissociation. Whereas the former is a painful but self-limiting injury with minimal sequelae, the latter is life threatening. However, the severity of most pectoral girdle injuries lies between these two extremes. Fractures and dislocations are common in this region throughout life, and a clear understanding of the patterns of injury and their radiographic spectrum is essential for all radiologists who deal with trauma. PMID- 11061700 TI - Fractures of the elbow and forearm. AB - Fractures and dislocations of the elbow usually occur secondary to indirect trauma. In the adult, fractures of the distal humerus almost always involve the condyles. Fractures of the radial head and neck may be subtle, and the appearance of secondary signs, such as the elevated fat pads from an elbow joint effusion, may be diagnostically useful. Dislocations of the elbow can be associated with fractures, such as those involving the ulnar coronoid process. In children, the presence of epiphyseal and apophyseal ossification centers can confuse the inexperienced observer in the setting of elbow trauma. Osteochondral injury may be difficult to identify without adjunctive imaging techniques, such as magnetic resonance (MR) imaging. Soft tissue injury at the elbow is also well characterized by MR imaging. Fractures of a single forearm bone may occur in isolation, usually due to a direct blow, but these are usually associated with fracture of displacement of the other bone in that forearm. PMID- 11061701 TI - Wrist and hand injuries. AB - Wrist and hand injuries are common occurrences. These are often caused by a fall on the outstretched hand, but other mechanisms, from sports-related injuries to high-speed motor vehicle collisions, can lead to injuries that may be subtle on initial imaging or may produce characteristic radiographic findings. In all cases, careful attention to certain anatomic details along with a sound understanding of different injury patterns should allow an accurate diagnosis to be made. The following this article illustrates these concepts, emphasizing features that should be familiar to both radiologists and orthopedic surgeons. PMID- 11061702 TI - Hip and femur trauma. AB - Fractures that involve the acetabulum and proximal femur are common. These injuries frequently occur due to high-energy trauma and may result in significant morbidity and long-standing disability, or high mortality when associated with other injuries, particularly to vascular structures. In this article, classifications pertinent to fractures of the acetabulum, hip dislocations, and fractures of the femoral neck, intertrochanteric region, and subtrochanteric region of the femur are discussed in detail with emphasis on their radiographic features. PMID- 11061703 TI - Injuries about the knee and tibial/fibular shafts. AB - Injuries to the knee and tibial/fibular shafts are extremely common, with knee injuries alone accounting for over 1.3 million emergency department visits yearly in the United States. Many of these injuries will present with straightforward radiographic findings, but others will have a subtle or complex appearance. This article reviews injuries of the knee and proximal tibial/fibular shaft with emphasis on normal anatomic features that, when disrupted, indicate the presence of subtle but important bone or soft tissue trauma. Although the emphasis is on plain radiography, magnetic resonance (MR) and computed tomographic (CT) findings are discussed where appropriate. PMID- 11061704 TI - Ankle and foot trauma. AB - This article reviews the most common foot and ankle injuries that the general radiologist and general orthopedic surgeon are likely to encounter. Fractures and dislocations seen in children and adults are included as well as soft tissue injuries. Ankle fracture classification schemes are discussed briefly. Techniques for advanced imaging with computed tomography (CT) and magnetic resonance (MR) are discussed where appropriate. PMID- 11061705 TI - Wound dressings reclassified by the FDA. PMID- 11061706 TI - Patient, wound, and treatment characteristics associated with healing in pressure ulcers. AB - OBJECTIVE: To examine the associations among selected patient, wound, and treatment factors and healing in pressure ulcers. DESIGN: Nonexperimental, retrospective study using data extracted from charts of patients having a Stage II, III, or IV pressure ulcer between July 1994 and November 1996. SETTING: A long-term-care facility associated with a major metropolitan medical center. RESULTS: Evidence of healing was significantly associated with patient weight (P < .05) and negatively associated with body temperature (P < .05), time on a pressure-relieving bed (P < .05), amount of exudate (P < .001), and stage of pressure ulcer (P < .001). In a regression analysis, pressure ulcer stage, patient weight, and mean body temperature explained 25% of the variability in healing; specifically, lower pressure ulcer stage, higher patient weight, and lower mean body temperature predicted improved healing. Healing was not predicted by chronologic age, pressure ulcer location, number of illnesses, use of tube feedings, use of pressure-relieving beds, mean arterial pressure, or Braden Scale score. In a regression analysis of patient factors alone, lower body temperature and higher weight together predicted 9% of the variance in healing. When wound variables were analyzed, 19% of the variability in healing was explained by the stage of the pressure ulcer. Of the treatment variables, only shorter time on a pressure-relieving bed predicted healing, explaining 6% of the variance. CONCLUSIONS: Strategies for healing pressure ulcers in nursing home patients should include programs for early recognition of pressure injury and prevention of pressure ulcer progression to higher stages; attention to weight gain and/or weight maintenance in at-risk patients; and early recognition and treatment of infections and febrile episodes. PMID- 11061707 TI - Osteomyelitis related to pressure ulcers: the cost of neglect. AB - Twelve patients with documented chronic osteomyelitis of the pelvis resulting from truncal pressure ulcers were examined retrospectively to identify the cost of treatment for this significant health care problem. The retrospective review of each case spanned an 18-month period--6 months prior to the initial positive bone biopsy to 1 year following bone biopsy. The financial charges associated with treatment of osteomyelitis were identified using the University of Michigan Health System's databases for hospital charges, professional charges, and pharmacy charges. Prior treatment of these patients included surgical debridement of the pressure ulcer, pelvic bone biopsy, and culture-specific antibiotic therapy. The total charges for this group of 12 patients was $715,204, or an average charge of $59,600 per patient. Each patient was hospitalized, with hospitalization charges of $587,212, or an average of $48,934 per patient. Pharmacy charges for culture-specific antibiotics totaled $85,217 for the 12 patients. Six of 8 flap repairs achieved successful surgical closure of the pressure ulcer (75%) postantibiotic therapy. Surgery charges are not included in the totals. PMID- 11061708 TI - Evaluating and managing the diabetic foot: an overview. PMID- 11061709 TI - Preventing diabetic foot complications. PMID- 11061710 TI - Wound characteristics by types. PMID- 11061711 TI - When to use hydrogel dressings. PMID- 11061712 TI - Assessment and treatment of involuntary weight loss and protein-calorie malnutrition. AB - Involuntary weight loss and protein-calorie malnutrition have a great impact on the entire health care system, resulting in reduced quality of life for the affected patient, compromised recovery, and added financial costs to the institution where the patient is receiving care. In addition, regulatory agencies are placing increased emphasis on the timely assessment and treatment of involuntary weight loss and protein-calorie malnutrition. In this context, a multidisciplinary approach to prevention of weight loss and protein-calorie malnutrition offers the greatest opportunity for success. Early recognition of protein-calorie malnutrition using anthropometric data, laboratory findings, and physical examination with a comprehensive diet history is essential to timely and effective treatment. Treatment must be closely monitored by a nutrition professional and must proceed along the accepted continuum of interventions. PMID- 11061713 TI - Involuntary weight loss and the nonhealing wound. AB - Wounds elicit a hypermetabolic, net catabolic state that can precipitate profound involuntary weight loss, particularly in the lean tissue compartment. To promote anabolic conditions more conducive to wound healing, an integrated treatment approach is warranted. Effective management includes optimal wound care, treatment or prevention of infection, nutritional intervention (e.g., protein supplementation), moderate exercise, and adjunctive anabolic treatment in patients whose involuntary weight loss has reached at least 10% of usual body weight. PMID- 11061714 TI - [Intravascular ultrasonographic evaluation of direct stents, implanted without predilatation. Comparison of results according to lesion types]. AB - INTRODUCTION AND OBJECTIVES: Direct stenting is a safe and feasible technique in selected lesions yielding excellent angiographic results. However, there are no studies providing intravascular ultrasonographic examination after direct stenting. The aim of this study was to evaluate direct stent expansion with ultrasonography and to know whether there are differences in the results based on lesion types. METHODS: Patients with amenable lesions for direct stenting were enrolled; including patients with no occlusion, no calcification, no significant tortuosity or angulation, a length 15 mm and a reference lumen diameter 2.5 mm. Intravascular ultrasonography was performed after stent implantation. The ultrasonographic criteria for optimal expansion were: complete apposition and a minimal intrastent lumen area > 80% of the average reference luminal area and 90% of the distal reference lumen area. RESULTS: We included 40 patients (50 lesions). The final angiographic result was good in all the patients but in one case an additional stent was used due to dissection. The ultrasonographic examination did not show significant differences between type A and B lesions. Optimal expansion was achieved in 14/21 (66%) of type A lesions and 17/29 (58%) of type B lesions (p = 0.5). The balloon/artery ratio was the only factor significantly related to ultrasonographic results. When this ratio was 1.1-1.2 (25 cases), 76% of the stents were optimally expanded and when the ratio was < 1. 1 (25 cases) only in a 48% an optimal result was achieved (p < 0.05). CONCLUSIONS: Direct stenting in selected lesions provides ultrasonographic results comparable to those expected with conventional stenting and these results could be even improved if a balloon artery ratio 1.1-1.2 is used. Taking into consideration the selection criteria the differences observed between lesion types A and B are not significant. PMID- 11061715 TI - [Clinical criteria to establish waiting times and prioritization for cardiovascular surgery. Official report of the Spanish Society of Cardiology and the Spanish Society of Cardiovascular Surgery]. PMID- 11061716 TI - The politics of risk: the case of BSE. If we want less paternalism we must first educate ourselves. PMID- 11061717 TI - Millennial musings. PMID- 11061718 TI - Clinical trials of medicines in children. PMID- 11061719 TI - Ten years of German unification. PMID- 11061720 TI - Taking heart failure seriously. PMID- 11061722 TI - In brief PMID- 11061721 TI - BSE inquiry plays down errors. PMID- 11061723 TI - Consent needed for organ retention, BMA says. PMID- 11061724 TI - Spain tops the table for organ donation. PMID- 11061725 TI - Celebrity illnesses raise awareness but can give wrong message. PMID- 11061726 TI - FDA panel advises against omeprazole as over the counter drug. PMID- 11061727 TI - Detection rates for breast cancer rising PMID- 11061728 TI - Where angels fear to tread. PMID- 11061729 TI - Bleeding and pneumonia in intensive care patients given ranitidine and sucralfate for prevention of stress ulcer: meta-analysis of randomised controlled trials. AB - OBJECTIVES: To determine the effectiveness of ranitidine and sucralfate in the prevention of stress ulcer in critical patients and to assess if these treatments affect the risk of nosocomial pneumonia. DESIGN: Published studies retrieved through Medline and other databases. Five meta-analyses evaluated effectiveness in terms of bleeding rates (A: ranitidine v placebo; B: sucralfate v placebo) and infectious complications in terms of incidence of nosocomial pneumonia (C: ranitidine v placebo; D: sucralfate v placebo; E: ranitidine v sucralfate). Trial quality was determined with an empirical ad hoc procedure. MAIN OUTCOME MEASURES: Rates of clinically important gastrointestinal bleeding and nosocomial pneumonia (compared between the two study arms and expressed with odds ratios specific for individual studies and meta-analytic summary odds ratios). RESULTS: Meta-analysis A (five studies) comprised 398 patients; meta-analysis C (three studies) comprised 311 patients; meta-analysis D (two studies) comprised 226 patients: and meta-analysis E (eight studies) comprised 1825 patients. Meta-analysis B was not carried out as the literature search selected only one clinical trial. In meta analysis A ranitidine was found to have the same effectiveness as placebo (odds ratio of bleeding 0.72, 95% confidence interval 0.30 to 1.70, P=0.46). In placebo controlled studies (meta-analyses C and D) ranitidine and sucralfate had no influence on the incidence of nosocomial pneumonia. In comparison with sucralfate, ranitidine significantly increased the incidence of nosocomial pneumonia (meta-analysis E: 1.35, 1.07 to 1.70, P=0.012). The mean quality score in the four analyses (on a 0 to 10 scale) ranged from 5.6 in meta-analysis E to 6.6 in meta-analysis A. CONCLUSIONS: Ranitidine is ineffective in the prevention of gastrointestinal bleeding in patients in intensive care and might increase the risk of pneumonia. Studies on sucralfate do not provide conclusive results. These findings are based on small numbers of patients, and firm conclusions cannot presently be proposed. PMID- 11061730 TI - Randomised, clinically controlled trial of intensive geriatric rehabilitation in patients with hip fracture: subgroup analysis of patients with dementia. AB - OBJECTIVE: To evaluate the effect of intensive geriatric rehabilitation on demented patients with hip fracture. DESIGN: Preplanned subanalysis of randomised intervention study. Settting: Jyvaskyla Central Hospital, Finland. PARTICIPANTS: 243 independently living patients aged 65 years or older admitted to hospital with hip fracture. INTERVENTION: After surgery patients in the intervention group (n=120) were referred to the geriatric ward whereas those in the control group were discharged to local hospitals. MAIN OUTCOME MEASURES: Length of hospital stay, mortality, and place of residence three months and one year after surgery for hip fracture. RESULTS: The median length of hospital stay of hip fracture patients with moderate dementia (mini mental state examination score 12-17) was 47 days in the intervention group (n=24) and 147 days in the control group (n=12, P=0.04). The corresponding figures for patients with mild dementia (score 18-23) were 29 days in the intervention group (n=35) and 46.5 days in the control group (n=42, P=0.002). Three months after the operation, in the intervention group 91% (32) of the patients with mild dementia and 63% (15) of the patients with moderate dementia were living independently. In the control group, the corresponding figures were 67% (28) and 17% (2). There were no significant differences in mortality or in the lengths of hospital stay of severely demented patients and patients with normal mini mental state examination scores. CONCLUSIONS: Hip fracture patients with mild or moderate dementia can often return to the community if they are provided with active geriatric rehabilitation. PMID- 11061731 TI - Circadian variation in onset of epistaxis: analysis of hospital admissions. PMID- 11061732 TI - Safety and costs of initiating angiotensin converting enzyme inhibitors for heart failure in primary care: analysis of individual patient data from studies of left ventricular dysfunction. AB - OBJECTIVE: To estimate the costs and consequences of diagnosing symptomatic heart failure with left ventricular systolic dysfunction and initiating angiotensin converting enzyme inhibitors in primary care. DESIGN: Analysis of individual patient data from studies of left ventricular dysfunction (SOLVD) to identify complications during test dose and titration phases. SETTING: Two randomised controlled trials in secondary care. PARTICIPANTS: 7487 patients taking a test dose of enalapril at enrolment to the treatment and prevention trials; 2569 patients with clinical signs of heart failure and established left ventricular dysfunction entered the treatment trial. MAIN OUTCOME MEASURES: Discontinuation during the test dose period. Discontinuation or reduction of dose during the first year of treatment for heart failure. Costs of diagnosis and titration of treatment. RESULTS: During the test dose phase, 585 patients (7.8%) reported side effects; 136 (1.8%) of these discontinued because of severe side effects. During the titration phase, compared with placebo, enalapril was associated with an increased risk of dose reduction due to hypotension (odds ratio 2.09, 95% confidence interval 1.15 to 3.82). However, overall, there was no difference in the rates of side effects leading to dose reduction or withdrawal between the enalapril and placebo groups. The costs of diagnosing heart failure with left ventricular systolic dysfunction and initiating and titrating an angiotensin converting enzyme inhibitor in primary care are pound300 to pound400. CONCLUSIONS: Treatment with angiotensin converting enzyme inhibitors can be safely started for patients with heart failure and left ventricular systolic dysfunction in primary care. PMID- 11061733 TI - Single gene disorders or complex traits: lessons from the thalassaemias and other monogenic diseases. PMID- 11061734 TI - ABC of colorectal cancer: primary treatment-does the surgeon matter? PMID- 11061735 TI - Poppy day again PMID- 11061736 TI - International comparators and poverty and health in Europe. PMID- 11061737 TI - Online patient-helpers and physicians working together: a new partnership for high quality health care. PMID- 11061738 TI - The role of complementary and alternative medicine. PMID- 11061739 TI - Broadening access to undergraduate medical education. PMID- 11061740 TI - Interprofessional education and teamworking: a view from the education providers. PMID- 11061741 TI - Teams: lessons from the world of sport. PMID- 11061742 TI - Will most people live in cities? PMID- 11061743 TI - Safeguards for research using large scale DNA collections. PMID- 11061744 TI - A healthy old age: realistic or futile goal? PMID- 11061745 TI - Cardiology: the development of a managed clinical network. PMID- 11061746 TI - Why a pomegranate? PMID- 11061747 TI - Decline in teenage smoking with rise in mobile phone ownership: hypothesis. PMID- 11061748 TI - Health hazards of mobile phones. Prevalence of headache is increased among users in Singapore. PMID- 11061749 TI - Health hazards of mobile phones. Do laptop computers also pose a risk? PMID- 11061750 TI - Anticoagulation to prevent stroke in atrial fibrillation. Rapid anticoagulant testing is not available in general practitioners' clinics in Japan. PMID- 11061751 TI - National claims database belongs to NHS as a whole. PMID- 11061752 TI - Reasons for inadequate health care vary. PMID- 11061753 TI - Informed consent is being neglected. PMID- 11061754 TI - RCGP should press for increase in resources. PMID- 11061755 TI - Parents do not always understand things doctors might say to them. PMID- 11061756 TI - Substance use seems to be increasing among 11 year olds. PMID- 11061757 TI - Earlobes are less convenient than fingerpricks for blood glucose testing. PMID- 11061758 TI - The new genetics: which genie out of which bottle? PMID- 11061759 TI - The role of task force junior doctor PMID- 11061760 TI - Surfactant treatment of neonates with respiratory failure and group B streptococcal infection. Members of the Collaborative European Multicenter Study Group. AB - OBJECTIVE: Connatal pneumonia caused by group B streptococcal (GBS) infection may be associated with surfactant dysfunction. We investigated the effects of surfactant treatment in term and preterm neonates with GBS infection and respiratory failure, in comparison with corresponding data from a control population of noninfected infants treated with surfactant for respiratory distress syndrome (RDS). DESIGN/METHODS: The study comprised 118 infants with respiratory failure, clinical and/or laboratory signs of acute inflammatory disease, and GBS infection proven by culture results. They were recruited retrospectively from a database of patients treated with surfactant at 28 neonatology units participating in European multicenter trials (1987-1993) and prospectively from the same units in the following years. A nonrandomized control group of 236 noninfected infants was selected from the same database. The primary parameters evaluated were oxygen requirement, ventilator settings, and incidence of complications. RESULTS: Median birth weight in the GBS study group was 1468 g (25th-75th percentiles: 1015-2170), and median gestational age was 30 (27-33) weeks. Thirty-one percent of the infants weighed >2000 g. Median age at surfactant treatment was 6 hours. The mean initial surfactant dose was 142 mg/kg (standard deviation: 53). Ninety of the infants were treated with Curosurf (Chiesi Farmaceutici, Parma, Italy), 13 with Survanta (Abboth GmbH, Wiesbaden, Germany), 12 with Alveofact (Dr Karl Thomae GmbH, Biberach, Germany), and 3 with Exosurf (Wellcome GmbH, Burgwedel, Germany). Within 1 hour of surfactant treatment, median fraction of inspiratory oxygen was reduced from .84 (25th-75th percentiles:.63-1.0) to.50 (.35-.80). The incidence of complications in the study group (mortality: 30%; pneumothorax: 16%; intracranial hemorrhage: 42%) was high, compared with infants with RDS. CONCLUSIONS: Surfactant therapy improves gas exchange in the majority of patients with GBS pneumonia. The response to surfactant is slower than in infants with RDS, and repeated surfactant doses are often needed. The mortality and morbidity are substantial, considering the relatively high mean birth weight of the treated infants. PMID- 11061761 TI - Safety and immunogenicity of heptavalent pneumococcal vaccine conjugated to CRM(197) among infants with sickle cell disease. Pneumococcal Conjugate Vaccine Study Group. AB - OBJECTIVES: To determine the immunogenicity and safety of heptavalent pneumococcal polysaccharide vaccine (serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F) conjugated to CRM(197) (7-valent conjugate pneumococcal vaccine [7VPnC]) among infants with sickle cell disease (SCD) and a comparison group of infants without SCD (non-SCD). DESIGN: Two cohorts of infants were enrolled and received open label doses of 7VPnC vaccine; infants enrolled before 2 months of age received 7VPnC vaccine at 2, 4, and 6 months of age followed by 23-valent pneumococcal polysaccharide vaccine (PS-23) at 24 months of age for those infants with SCD (schedule A), and infants enrolled between 2 and 12 months of age received 7VPnC at 12 months of age followed by PS-23 at 24 months of age for infants with SCD (schedule B). Safety data were collected for 3 days after each dose of vaccine. Antibody concentrations were measured to each of the 7VPnC serotypes by enzyme linked immunosorbent assay before each vaccine dose and 1 month after the last 7VPnC dose and the PS-23 vaccine dose. RESULTS: Forty-five infants (34 SCD and 11 non-SCD) were vaccinated according to schedule A and 16 infants (13 SCD and 3 non SCD) according to schedule B. The 7VPnC vaccine was highly immunogenic for all serotypes among infants with and without SCD who received 3 doses of vaccine according to schedule A: depending on serotype, 89% to 100% achieved antibody concentrations above.15 microg/mL and 56% to 100% achieved antibody concentrations above 1.0 microg/mL. Among infants immunized according to schedule B, a single dose of 7VPnC vaccine resulted in antibody concentrations above.15 microg/mL in 53% to 92% by serotype and above 1.0 microg/mL in 31% to 71% by serotype. A single dose of PS-23 resulted in dramatic increases in the antibody concentrations to all serotypes regardless of 1- or 3-dose priming. There was no difference in the reactogenicity of the 7VPnC vaccine between those with and without SCD. There were no serious reactions to the 7VPnC or PS-23 vaccines, even among those with high antibody concentrations before immunization. CONCLUSIONS: Infants with SCD respond to 7VPnC vaccine with antibody concentrations that are at least as high as infants without SCD. Infants immunized with 7VPnC vaccine at 2, 4, and 6 months of age developed antibody concentrations in the same range as those achieved among infants without SCD enrolled in a large trial that demonstrated vaccine efficacy against invasive disease. Significant rises were seen in antibody concentrations to all 7VPnC serotypes after the PS-23 booster in children receiving schedule A or B. PMID- 11061762 TI - Economic impact of influenza vaccination in preschool children. AB - OBJECTIVE: The economic impact of routine vaccination of preschool children with inactivated influenza vaccine was investigated. DESIGN: A decision analysis was performed using data from the literature. Direct and indirect costs of each vaccination strategy were calculated and compared with a strategy of not vaccinating. SETTING: Two settings were evaluated: a setting in which vaccination was available during flexible hours and a setting in which vaccination was available only during usual work hours (8:00 am-5:00 pm). RESULTS: Vaccination resulted in a net cost savings in both settings. The net savings per vaccine recipient were $21.28 in the flexible setting and $1.20 in the restricted setting. Although the analysis was performed for the inactivated vaccine, sensitivity analysis showed that the nasal vaccine could also result in a net cost savings depending on the price of the cold-adapted vaccine when it is licensed. CONCLUSION: Vaccinating preschool children is economically advantageous. Serious consideration should be given to recommending vaccination in this age group. PMID- 11061763 TI - Predictors of bacteremia in febrile children 3 to 36 months of age. AB - PURPOSE: To develop an improved model for the prediction of bacteremia in young febrile children. METHODS: A retrospective review was performed on patients 3 to 36 months of age seen in a children's hospital emergency department between December 1995 and September 1997 who had a complete blood count and blood culture ordered as part of their regular care. Exclusion criteria included current use of antibiotics or any immunodeficient state. Clinical and laboratory parameters reviewed included age, gender, race, weight, temperature, presence of focal bacterial infection, white blood cell count (WBC), polymorphonuclear cell count (PMN), band count, and absolute neutrophil count (ANC). Logistic regression analyses were used to identify factors associated with bacteremia, defined as growth of a pathogen in a blood culture. The model that was developed was then validated on a second dataset consisting of febrile patients 3 to 36 months of age collected from a second children's hospital (validation set). RESULTS: There were 633 patients in the derivation set (46 bacteremic) and 9465 patients in the validation set (149 bacteremic). The mean age of patients in the derivation and validation sets were 15.8 months (95% confidence interval [CI]: 15.2-16.5) and 16.6 months (95% CI: 16.5-16.8), respectively; the mean temperatures were 39.1 degrees C (95% CI: 39. 0-39.2) and 39.8 degrees C (95% CI: 39.7-39.8); 56% were male in the derivation set and 55% male in the validation set. Predictors of bacteremia identified by logistic regression included ANC, WBC, PMN, temperature, and gender. Receiver operator characteristic (ROC) analysis showed similar performance of ANC and WBC as predictors of bacteremia. When placed into a multivariate logistic regression model, band count was not significantly associated with bacteremia. Information regarding focal infection was available for 572 patients in the derivation set. The percentage of patients diagnosed with bacteremia with a focal bacterial infection was not significantly different from the percentage who had bacteremia without a focal bacterial infection (16/200 vs 30/372). Based on this dataset, a logistic regression formula was developed that could be used to develop a unique risk value for each patient based on temperature, gender, and ANC. When the final model was applied to the validation set, the area under the ROC curve (AUC) constructed from these data indicated that the model retained good predictive value (AUC for the derivation vs validation data =.8348 vs 0.8221, respectively). CONCLUSIONS: Use of the formulas derived here allows the clinician to estimate a child's risk for bacteremia based on temperature, ANC, and gender. This approach offers a useful alternative to predictions based on fever and WBC alone.bacteremia, detection, white blood cell. PMID- 11061764 TI - Intrapartum fever and unexplained seizures in term infants. AB - OBJECTIVE: Early-onset neonatal seizures are a strong predictor of later morbidity and mortality in term infants. Although an association of noninfectious intrapartum fever with neonatal seizures in term infants has been reported, it was based on only a small number of neonates with seizures. We therefore conducted a case control study to investigate this association further. METHODS: All term infants with neonatal seizures born at Brigham and Women's Hospital between 1989 and 1996 were identified. For this study, cases consisted of all term neonates with a confirmed diagnosis of seizure born after a trial of labor for whom no proximal cause of seizure could be identified. Infants with sepsis or meningitis were excluded. Four controls matched by parity and date of birth were identified for each case. The rate of intrapartum maternal temperature >100.4 degrees F was compared for case infants and controls. Potential confounding was controlled in logistic regression analysis. RESULTS: Cases comprised 38 term infants with unexplained seizures after a trial of labor. We identified 152 controls. Infants with seizures were more likely to be born to mothers who were febrile during labor (31.6% vs 9.2%). In almost all cases, the fever developed during labor (94.7% cases, 97.4% controls). At admission, mothers of infants with seizures were not significantly more likely to have factors associated with concern about infection such as a white blood cell count >15 000/mm(3) (28. 9% vs 19.1%) and premature rupture of the membranes (15.8% vs 17.8%). In a logistic regression analysis controlling for confounding factors, intrapartum fever was associated with a 3.4-fold increase in the risk of unexplained neonatal seizures (odds ratio = 3.4, 95% confidence interval = 1.03-10.9). CONCLUSION: Our data indicate that intrapartum fever, even when unlikely to be caused by infection, is associated with a fourfold increase in the risk of unexplained, early-onset seizures in term infants. PMID- 11061765 TI - A randomized, double-blind, placebo-controlled study of sumatriptan nasal spray in the treatment of acute migraine in adolescents. AB - OBJECTIVE: To compare the efficacy and tolerability of sumatriptan nasal spray (NS; 5 mg, 10 mg, and 20 mg) with placebo for the treatment of acute migraine in adolescents. METHODS: A randomized, double-blind, placebo-controlled, single attack study was conducted in 653 US adolescents (12-17 years of age). Patients with at least a 6-month history of migraine, who met International Headache Society criteria for migraine (with or without aura) were eligible for participation. Headache relief 2 hours postdose, complete relief, presence or absence of associated symptoms, headache recurrence, and use of rescue medications were recorded. The primary efficacy endpoint was headache relief 2 hours postdose sumatriptan NS (20 mg) versus placebo. Safety and tolerability were assessed by examining adverse events, changes in electrocardiograms, vital signs, physical examinations, and clinical laboratory tests. RESULTS: Headache relief 1 hour postdose was significantly greater for patients using 10 mg (56%) and 20 mg (56%) of sumatriptan NS compared with placebo (41%). Headache relief 2 hours postdose was significantly greater for patients using 5 mg of sumatriptan NS (66%) compared with placebo (53%), and approached statistical significance for 20 mg (63%) compared with placebo (53%). Complete relief 2 hours postdose was significantly greater for patients using 20 mg of sumatriptan NS compared with placebo (36% vs 25%, respectively). Each dose of sumatriptan (5 mg, 10 mg, and 20 mg) was superior to placebo with respect to the cumulative percentages of patients first reporting headache relief within 2 hours of dosing (Kaplan-Meier). The sumatriptan 20-mg dose was superior to placebo with respect to the cumulative percentages of patients first reporting complete relief within 2 hours of dosing (Kaplan-Meier). Photophobia and phonophobia were significantly reduced 2 hours postdose for sumatriptan NS (20 mg), compared with placebo (36% vs 48% and 25% vs 44%, respectively). Taste disturbance was the most commonly reported adverse event (2%, 19%, 30%, and 26% for placebo, 5 mg, 10 mg, and 20 mg, respectively). No drug-related serious adverse events or clinically relevant changes in laboratory parameters, electrocardiograms, or vital signs were reported. CONCLUSIONS: Sumatriptan NS is effective and well-tolerated for the treatment of acute migraine in adolescents, with the 20-mg dose providing the best overall efficacy and tolerability profiles. PMID- 11061766 TI - Severity of neonatal retinopathy of prematurity is predictive of neurodevelopmental functional outcome at age 5.5 years. Behalf of the Cryotherapy for Retinopathy of Prematurity Cooperative Group. AB - OBJECTIVE: The purpose of this study was to assess the relation between neonatal retinopathy of prematurity (ROP) in very low birth weight infants and neurodevelopmental function at age 5.5 years. METHODS: Longitudinal follow-up of children occurred in 2 cohorts of the Multicenter Cryotherapy for Retinopathy of Prematurity Study. The extended natural history cohort followed 1199 survivors of <1251 g birth weight from 5 centers. The threshold randomized cohort (ThRz) followed 255 infants <1251 g from 23 centers who developed threshold ROP and who consented to cryotherapy to not more than 1 eye. At 5.5 years both cohorts had ophthalmic and acuity testing and neurodevelopmental functional status determined with the Functional Independence Measure for Children (WeeFIM). RESULTS: Evaluations were completed on 88.7% of the extended natural history cohort; 87% had globally normal functional skills (WeeFIM: >95). As ROP severity increased, rates of severe disability increased from 3.7% among those with no ROP, to 19.7% of those with threshold ROP. Multiple logistic regression analysis demonstrated that better functional status was associated with favorable visual acuity, favorable 2-year neurological score, absence of threshold ROP, having private health insurance, and black race. Evaluations were completed on 87.4% of the ThRz children. In each functional domain, the 134 children with favorable acuity in their better eye had fewer disabilities than did the 82 children with unfavorable acuity: self-care disability 25.4% versus 76.8%, continency disability 4.5% versus 50.0%, motor disability 5.2% versus 42.7%, and communicative-social cognitive disability 22.4% versus 65.9%, respectively. CONCLUSION: Severity of neonatal ROP seems to be a marker for functional disability at age 5. 5 years among very low birth weight survivors. High rates of functional limitations in multiple domains occur in children who had threshold ROP, particularly if they have unfavorable visual acuity. PMID- 11061767 TI - Effectiveness of a clinical pathway for inpatient asthma management. AB - BACKGROUND: Clinical pathways for asthma are tools that have the potential to improve compliance with nationally recognized management guidelines, but their effect on patient outcomes has not been documented. OBJECTIVES: To determine the effect of an asthma clinical pathway on patients' length of stay, use of nebulized beta-agonist therapy while hospitalized, and use of acute care clinics for 2 weeks after discharge. DESIGN/METHODS: The study was a randomized, controlled trial. Patients between the ages of 2 and 18 years admitted with an asthma exacerbation and not under the care of an asthma specialist were eligible for the study. Patients were randomized either to a conventional ward (control group) or to a ward using the clinical pathway (intervention group). For 2 weeks after discharge, we collected data to determine whether patients visited a health care provider for worsening asthma. RESULTS: One hundred ten patients (26%) were enrolled. Control and intervention groups had similar demographic and asthma severity profiles. The intervention group had an average length of stay 13 hours shorter than did the control group. In addition, at every dosing interval, the intervention group received less nebulized beta-agonist therapy. There were no deaths in either group. CONCLUSION: A clinical pathway for inpatient asthma decreased the length of stay and beta-agonist medication use with no adverse outcomes or increased acute-care encounters through 2 weeks after discharge. PMID- 11061768 TI - Respiratory symptoms in mothers of young children. AB - OBJECTIVES: Children receiving child care outside the home are at greater risk of upper respiratory infection, but whether parents of those children are also at increased risk is undocumented. We describe the incidence of 2 or more respiratory symptoms in the previous 2 weeks among 185 mothers of children 3 years of age or younger by child care use. METHODS: Mothers in Michigan and Nebraska were interviewed by phone regarding respiratory symptoms, use of outside child care (for an index child), sleeping habits, and demographic information. RESULTS: Nearly one half (46.5%) reported 2 or more symptoms during the past 2 weeks; 15.1% had contacted a health care provider and 13.0% spent 1 or more days in bed because of their symptoms, which lasted an average of 5.5 days. Prevalence of symptoms was invariant to sociodemographic characteristics. Mothers using outside child care (74.6%) were twice as likely as those without outside care to have been ill in the past 2 weeks (odds ratio: 2.26; 95% confidence interval [CI]: 1.12,4.54). Most mothers (69.2%) reported having their sleep interrupted by their children at least once in the last 2 weeks or sharing a bed with a child part or all of the night (61.1%); 25.4% slept 6 hours or less nightly. Women reporting that they rarely or never felt rested (26. 5%) were 2.65 times more likely to be ill (95% CI: 1.26,5.55), compared with those reporting that they frequently or always felt rested (46.5%), after adjusting for any outside child care. CONCLUSIONS: Future studies should focus on risk factors that can be modified to reduce illness among both children and their parents. PMID- 11061769 TI - School disconnectedness: identifying adolescents at risk. AB - OBJECTIVE: School connectedness, or the feeling of closeness to school personnel and the school environment, decreases the likelihood of health risk behaviors during adolescence. The objective of this study was to identify factors differentiating youth who do and do not feel connected to their schools in an effort to target school-based interventions to those at highest health risk. METHODS: The study population consisted of all students attending the 7th through 12th grades of 8 public schools. The students were asked to complete a modified version of the in-school survey designed for the National Longitudinal Study of Adolescent Health (Add Health). The school connectedness score (SCS) was the summation of 5 survey items. Bivariate analyses were used to evaluate the association between SCS and 13 self-reported variables. Stepwise linear regression was conducted to identify the set of factors best predicting connectedness, and logistic regression analysis was performed to identify students with SCS >1 standard deviation below the mean. RESULTS: Of the 3491 students receiving surveys, 1959 (56%) submitted usable surveys. The sample was 47% white and 38% black. Median age was 15. Median grade was 9th. The SCS was normally distributed with a mean of 15.7 and a possible range of 5 to 25. Of the 12 variables associated with connectedness, 7 (gender, race, extracurricular involvement, cigarette use, health status, school nurse visits, and school area) entered the linear regression model. All but gender were significant in the logistic model predicting students with SCS >1 standard deviation below the mean. CONCLUSIONS: In our sample, decreasing school connectedness was associated with 4 potentially modifiable factors: declining health status, increasing school nurse visits, cigarette use, and lack of extracurricular involvement. Black race, female gender, and urban schools were also associated with lower SCS. Further work is needed to better understand the link between these variables and school connectedness. If these associations are found in other populations, school health providers could use these markers to target youth in need of assistance. PMID- 11061770 TI - Care to underserved children: Residents' attitudes and experiences. AB - OBJECTIVES: To examine: 1) the relationship between residents' responses toward caring for underserved children and families during residency and their perceptions of their continuity clinic experience; and 2) which characteristics are related to continuing to care for the underserved after completion of training. DESIGN: Cross-sectional descriptive study. METHODS: A 49-item questionnaire was mailed to 210 third-year pediatric residents at 12 urban training programs in the Northeast in May 1995. Information was collected about residents' emotional responses toward caring for underserved families, their assessments of clinic operations, their sense of effectiveness in caring for underserved patients in continuity clinic, preresidency experiences with the underserved, and their intent to care for the underserved after training. RESULTS: Of 210 surveys mailed, 71% were returned. Thirty-six percent of residents planned to pursue a career in primary care, 53% did not, and 11% did not answer or were planning a year as chief resident. Fifty-seven percent of all residents planned to devote a portion of their practice toward caring for the underserved after training. Residents whose emotional responses toward caring for the underserved included: 1) not worrying that they had become numb to children's psychosocial difficulties, 2) not feeling angry with how families cared for their children, and 3) feeling more empathy with the underserved had significantly higher mean scores on both their assessment of clinic operations and their sense of effectiveness. The only demographic characteristic associated with a greater sense of effectiveness was being black. To better characterize which residents planned to care for the underserved after training, we examined a subsample of 46 residents who recalled an interest in caring for the underserved during residency training and who were pursuing a career in primary care. Residents that did not recall an interest in caring for the underserved at the onset of residency training were unlikely to have plans to care for the underserved after the completion of training. Within this group residents who planned to care for the underserved after training differed significantly from residents who did not plan to continue this work by feeling a greater sense of effectiveness in clinic, feeling less worried about becoming numb, and having greater empathy for underserved families. CONCLUSIONS: There are a number of identifiable emotional responses residents develop toward caring for the underserved that relate to their perceptions of continuity clinic and whether a resident chooses to continue to care for the underserved after training. PMID- 11061771 TI - Cisapride and QTc interval in children. AB - BACKGROUND: Recent reports about cisapride have raised some concerns about the safety and efficacy of this medication in children. The aim of this study was to identify electrocardiographic changes and a predisposition to develop arrhythmias in children. METHODS: Patients were divided in 2 groups: 1) 63 children (mean age: 29 months) who received cisapride (0.2 mg/kg/dose 3 times/day), and 2) 57 children (mean age: 27 months) who did not receive cisapride (they served as controls). Both groups did not have any associated disease. Electrocardiogram (EKG) was performed to children when they were included in the study. The QT interval was corrected using Bazett's formula. Twenty-four-hour Holter recording was performed in children with prolonged QT interval (PQTI). When PQTI was identified in group 1, cisapride was discontinued and a new EKG was performed. RESULTS: Five children from group 1 and 6 from group 2 had PQTI. In 3 children with PQTI, the QTc interval returned to normal values when cisapride was discontinued. In children under 4 months of age, a statistical difference was found, with QTc interval being longer in group 2 (without cisapride) than in group 1. Holter recordings were normal in all children with PQTI. CONCLUSION: PQTI can be found in normal children with or without cisapride. In our study PQTI was not associated with any life-threatening event. PMID- 11061772 TI - Factors influencing infant visits to emergency departments. AB - OBJECTIVES: To follow the 1995 birth cohort of infants, born in the State of Missouri, through their first birthday to: 1) examine their rates of visits to emergency departments (EDs), 2) identify predictors of any ED visit, 3) examine rates of nonurgent ED visits, and 4) identify predictors of nonurgent visits. METHODS: This was a retrospective population cohort study. Using deterministic linkage procedures, 2 databases at the Missouri Department of Health (DOH; (the patient abstract database and the birth registry database) were linked by DOH personnel. International Classification of Diseases, Ninth Revision-Clinical Modification codes for ED visits were classified as emergent, urgent, or nonurgent by 2 researchers. Eight newborn characteristics were chosen for analysis. Negative binomial regression was used to examine the rates and predictors of both total and nonurgent ED visits. RESULTS: There were 935 total ED visits and 153 nonurgent ED visits per 1000 infant years. The average number of visits was.94, with 59% of infants having no visits, 21% having 1 ED visit, and 20% having 2 or more visits. Factors associated with increases in both total and nonurgent ED visits were Medicaid, self-pay, black race, rural region, presence of birth defects, and a nursery stay of >2 days. Significant interactions were found between Medicaid and race and Medicaid and rural regions on rates of ED use and nonurgent use. The highest rate of ED use, 1.8 per person year, was seen in white, rural infants on Medicaid, and the lowest rate (.4 per person year) was seen in urban white infants not on Medicaid. The highest rates of nonurgent use,.3 per person year, were among urban and rural Medicaid infants of both races and among black infants on commercial insurance. The lowest nonurgent rate,.04 per person year, was seen in white urban infants on commercial insurance. CONCLUSION: Infants in the State of Missouri have high rates of ED visits. Nonurgent visits are only a small portion of ED visits and cannot explain large variations in ED usage. Increased ED use by Medicaid patients may reflect continuing difficulties in accessing primary care. PMID- 11061773 TI - Can epinephrine inhalations be substituted for epinephrine injection in children at risk for systemic anaphylaxis? AB - BACKGROUND: For out-of-hospital treatment of anaphylaxis, inhalation of epinephrine from a pressurized metered-dose inhaler is sometimes recommended as a noninvasive, user-friendly alternative to an epinephrine injection. OBJECTIVE: To determine the feasibility of administering an adequate epinephrine dose from a metered-dose inhaler in children at risk for anaphylaxis by assessing the rate and extent of epinephrine absorption after inhalation. METHODS: We performed a prospective, randomized, observer-blind, placebo-controlled, parallel-group study in 19 asymptomatic children with a history of anaphylaxis. Based on the child's weight, 10, 15, or 20 carefully supervised epinephrine or placebo inhalations were attempted. Before dosing, and at intervals from 5 to 180 minutes after dosing, we monitored plasma epinephrine concentrations, blood glucose, heart rate, blood pressure, and adverse effects. RESULTS: Eleven children (mean +/- standard error of the mean: 9 +/- 1 years and 33 +/- 3 kg) in the epinephrine group were able to inhale 11 +/- 2 (range: 3-20) puffs, equivalent to 74% +/- 7% of the precalculated dose or 0.078 +/- 0.009 mg/kg. They achieved a mean peak plasma epinephrine concentration of 1822 +/- 413 (range: 230-4518) pg/mL at 32.7 +/- 6.2 minutes. Eight children (10 +/- 1 years of age and 33 +/- 5 kg) in the placebo group were able to inhale 12 +/- 2 (range: 8-20) puffs, 89% +/- 3% of the precalculated dose, and had a peak endogenous plasma epinephrine concentration of 1316 +/- 247 (range: 522-2687) pg/mL at 44.4 +/- 16.7 minutes. In the children receiving epinephrine compared with those receiving placebo, mean plasma epinephrine concentrations were not significantly higher at any time, mean blood glucose concentrations were significantly higher from 10 to 30 minutes, mean heart rate was not significantly different at any time, and mean systolic and diastolic blood pressures were not significantly increased at most times. After the inhalations of epinephrine or placebo, the children complained of bad taste and many experienced cough or dizziness. After inhaling epinephrine, 1 child developed nausea, pallor, and muscle twitching. CONCLUSIONS: Despite expert coaching, because of the number of epinephrine inhalations required and the bad taste of the inhalations, most children were unable to inhale sufficient epinephrine to increase their plasma epinephrine concentrations promptly and significantly. Therefore, we urge caution in recommending epinephrine inhalation as a substitute for epinephrine injection for out-of-hospital treatment of anaphylaxis symptoms in children. PMID- 11061774 TI - Primitive neuroectodermal tumors of the brainstem: investigation of seven cases. AB - OBJECTIVE: We discuss the clinical aspects, pathology, and molecular genetics of 7 patients with primitive neuroectodermal tumors (PNETs) arising in the brainstem that were treated at our institution from 1986 through 1995. Most neuro oncologists avoid performing biopsies in children with pontine tumors. This article raises the question as to whether biopsies should be performed, because treatment recommendations might differ if a PNET was diagnosed rather than a pontine glioma. PATIENTS AND METHODS: We reviewed the clinical neuro-oncology database and the files of the Division of Neuropathology at New York University Medical Center from 1986 through 1995 and identified 7 histologically confirmed PNETs arising in the brainstem among 146 pediatric brainstem tumors. The clinical, neuroradiological, and neuropathological data were reviewed. Postmortem examinations were performed in 2 cases. Formalin-fixed, paraffin-embedded tumor tissues were also available in 6 of 7 patients that were tested for p53 gene mutations using single-strand conformation polymorphism analysis. We also tested 9 cerebellar PNETs, 9 brainstem gliomas, and 3 normal brains for p53 gene mutations as controls. RESULTS: All 7 patients presented with focal cranial nerve deficits, and 2 were also hemiparetic. The median age at diagnosis was 2.7 (1-8 years). Magnetic resonance imaging (MRI) characteristics included a focal intrinsic exophytic nonenhancing brainstem lesion that had low T1-weighted and high T2-weighted signals. Hydrocephalus was present in 5 patients at diagnosis, 3 of whom had leptomeningeal dissemination. Meningeal dissemination occurred later in the course of the disease in 3 other patients. Five children required shunts at diagnosis and another 2 at recurrence. Despite therapy, all 7 PNET patients died within 17 months of diagnosis with a mean survival of 8 (4-17) months. No mutation in the p53 gene was detected. CONCLUSIONS: Brainstem PNETs tend to arise at a younger age than brainstem gliomas and medulloblastomas. The MRI pattern suggests a localized rather than a diffuse intrinsic nonenhancing brainstem tumor. Like other PNETs, brainstem PNETs have a high predilection to disseminate within the central nervous system. The absence of p53 mutations is similar to other PNETs. Despite their origin close to the cerebellum, brainstem PNETs exhibit a more aggressive behavior and result in worse clinical outcomes than do cerebellar PNETs. PMID- 11061775 TI - The Child-Friendly Healthcare Initiative (CFHI): Healthcare provision in accordance with the UN Convention on the Rights of the Child. Child Advocacy International. Department of Child and Adolescent Health and Development of the World Health Organization (WHO). Royal College of Nursing (UK). Royal College of Paediatrics and Child Health (UK). United Nations Children's Fund (UNICEF). AB - OBJECTIVE: Although modern medical technology and treatment regimens in well resourced countries have improved the survival of sick or injured children, most of the world's families do not have access to adequate health care. Many hospitals in poorly resourced countries do not have basic water and sanitation, a reliable electricity supply, or even minimal security. The staff, both clinical and nonclinical, are often underpaid and sometimes undervalued by their communities. In many countries there continues to be minimal, if any, pain control, and the indiscriminate use of powerful antibiotics leads to a proliferation of multiresistant pathogens. Even in well-resourced countries, advances in health care have not always been accompanied by commensurate attention to the child's wider well-being and sufficient concerns about their anxieties, fears, and suffering. In accordance with the United Nations Convention on the Rights of the Child,(1) the proposals set out in this article aim to develop a system of care that will focus on the physical, psychological, and emotional well-being of children attending health care facilities, particularly as inpatients. DESIGN OF THE PROGRAM: To develop in consultation with local health care professionals and international organizations, globally applicable standards that will help to ensure that practices in hospitals and health centers everywhere respect children's rights, not only to survival and avoidance of morbidity, but also to their protection from unnecessary suffering and their informed participation in treatment. Child Advocacy International will liase closely with the Department of Child and Adolescent Health and Development of the World Health Organization (WHO) and the United Nations Children's Fund (UNICEF) in the implementation of the pilot scheme in 6 countries. In hospitals providing maternity and newborn infant care, the program will be closely linked with the Baby Friendly Hospital Initiative of WHO/UNICEF that aims to strengthen support for breastfeeding. United Nations Children's Fund, United Nations Convention on the Rights of the Child, child protection, breastfeeding, pain control, palliative care, child abuse. PMID- 11061776 TI - Using a health concerns checklist as a bridge from reason for encounter to diagnosis of girls attending an adolescent health service. AB - OBJECTIVE: We assessed the extent to which a health concerns checklist (HCC) helps bridge the gap between the reason for encounter (RFE) described by girls entering an adolescent health service and the ultimate diagnosis. METHODS: The sample, 547 consecutive 12- to 18-year-old girls visiting an adolescent health service, first underwent a structured intake procedure, including a self administered form on which they described their RFEs and other health concerns, as well as a psychosocial interview and medical evaluation performed by staff members. The RFEs, HCC items, and diagnoses, grouped into somatic, sexuality related, and psychosocial categories, were then compared. RESULTS: Among the 399 girls expressing specific RFEs on entering the clinic, one-third were diagnosed with psychosocial disorders and one-fifth with sexuality-related concerns. Of the patients receiving a sexuality-related diagnosis, 57% presented with a sexuality related request; another 26% noted it on the checklist. For those diagnosed with psychosocial problems, 22% stated this as the RFE, and another 50% indicated it on the HCC. The contribution of the HCC to the diagnosis was higher among adolescents not stating a specific RFE. CONCLUSION: The findings highlight the HCC's contribution in identifying health problems, especially among adolescents who find it difficult to verbalize sensitive issues. PMID- 11061777 TI - Variations in practice and outcomes in the Canadian NICU network: 1996-1997. AB - BACKGROUND: Previous reports of variations in outcomes among neonatal intensive care units (NICUs) examined only specific subpopulations of interest (eg, very low birth weight [VLBW] infants <1500 g of birth weight [BW]). OBJECTIVES: We report on current practice and outcomes variations in a population-based national study of Canadian NICUs from January 8, 1996 to October 31, 1997. METHOD: Information on 20 488 admissions to 17 tertiary level NICUs across Canada was prospectively collected by trained abstractors using a standard manual of operations and definitions. Data were verified and analyzed in concert with a steering committee comprising experienced researchers and neonatologists. Patient information included demographic information, antenatal history, mode of delivery, problems at delivery, status of infant and problems at birth, illness severity (Clinical Risk Index for Babies, Score for Neonatal Acute Physiology, Score for Neonatal Acute Physiology-Version II), therapeutic intensity (Neonatal Therapeutic Intensity Scoring System [NTISS]), selected NICU practices and procedures, use of technology and resources, and selected patient outcomes. Patients were tracked until death or discharge home. RESULTS: The mean number of annual admissions to an NICU was 657, with 26% outborn infants. Fifty-three percent were <2500 g BW, 20% were <1500 g BW (VLBW), and 65% were preterm (<38 weeks' gestational age [GA]). Only 2% of mothers received no prenatal care. Antenatal steroids were given to 58%, but there was wide variation in use (23% 76%). Congenital anomalies were present in 14%, and 4% were small for GA (less than the third percentile). Admission illness severity was lowest among infants 33 to 37 weeks of GA and correlated with risk of death. Ninety-six percent of patients survived until discharge, but fewer survived at lower GA. No infant <22 weeks' GA survived. Seven percent of infants had at least 1 episode of infection, but 75% received antibiotics in the NICU. Forty-three percent received respiratory support, and 14% received surfactant. Nitric oxide was given to 150 term infants and to 102 preterm infants. Selected outcomes of VLBW infants were: survival rate (87%); chronic lung disease (26%); >/=stage 3 retinopathy of prematurity (ROP; 11%); >/=grade 3 intraventricular hemorrhage (IVH; 10%); nosocomial infection (22%); necrotizing enterocolitis (NEC; 7%). Sixty-nine percent of VLBW infants survived without major morbidity (>/=grade 3 IVH, chronic lung disease, NEC, >/=grade 3 ROP). The mean duration of NICU stay was 19 days. Forty-seven percent of infants were discharged from the hospital, and 43% were retrotransferred to a community facility before discharge home. Significant variation in practices and outcomes were observed in all aspects of NICU care. CONCLUSION: This study provides population-based information about NICU outcomes. Significant variation in NICU practices and outcomes was observed despite Canada's universal health insurance system. This national database provides valuable information for planning research, allocating resources, designing health and public policy, and serving as a basis for longitudinal studies of NICU care in Canada. PMID- 11061778 TI - Necrotizing enterocolitis in neonates with congenital heart disease: risk factors and outcomes. AB - OBJECTIVE: Necrotizing enterocolitis (NEC) is primarily a disease of the premature infant. Among children born at term, however, congenital heart disease may be an important predisposing factor for this condition. To determine risk factors for NEC in patients with congenital heart disease, we conducted a case control study of neonates with cardiac disease admitted to the cardiac intensive care unit at our center during the 4-year period from January 1995 to December 1998. METHODS: Cardiac diagnosis and age at admission were analyzed for association with NEC among the 643-patient inception cohort. Demographic, preoperative, and operative variables were recorded retrospectively in 21 neonates with congenital heart disease who developed NEC and 70 control neonates matched by diagnosis and age at admission. Using parametric and nonparametric analysis, cases and controls were compared with respect to previously identified risk factors for NEC. RESULTS: Among the entire cohort of 643 neonates with heart disease admitted to the cardiac intensive care unit, diagnoses of hypoplastic left heart syndrome (odds ratio [OR] = 3.8 [1.6-9.1]) and truncus arteriosus or aortopulmonary window (OR = 6.3 [1.7-23.6]) were independently associated with development of NEC by multivariable analysis. In the case-control analysis, earlier gestational age at birth (36.7 +/- 2. 7 weeks vs 38.1 +/- 2.3 weeks), prematurity (OR = 3.9 [1.2-12.5]), highest dose of prostaglandin >0.05 microg/kg/minute (OR = 3.9 [1. 2-12.5]), and episodes of low cardiac output (meeting specific laboratory criteria) or clinical shock (OR = 6.5 [1.8-23.5]) correlated with the development of NEC. Earlier gestational age and episodes of low output were the only factors that remained significantly associated with NEC by multivariable analysis. Although there was no difference in hospital mortality between patients with and without NEC, mean hospital stay was significantly longer in those who developed NEC (36 +/- 22 days vs 19 +/- 14 days). CONCLUSIONS: The risk of NEC in neonates with congenital heart disease is substantial. Factors associated with an elevated risk of NEC in infants with heart disease include premature birth, hypoplastic left heart syndrome, truncus arteriosus, and episodes of poor systemic perfusion or shock. Heightened suspicion is warranted in newborns with these risk factors. PMID- 11061779 TI - Daily physical activity program increases bone mineralization and growth in preterm very low birth weight infants. AB - OBJECTIVE: A study of daily physical activity was performed with 32 preterm infants to evaluate changes in body weight and bone mineralization. STUDY DESIGN: Subjects were matched by birth weight and gestational age and randomly assigned to the physical activity (PA; n = 16) or to the control (C; n = 16) program. PA consisted of range of motion against passive resistance to all extremities for 5 to 10 minutes daily. Peripheral dual-energy x-ray of the right forearm (ulna and radius); biomarkers of bone formation (serum type I collagen C-terminal propeptide [PICP]) and resorption (urine pyridinoline cross-links of collagen [Pyd]); serum calcium, phosphate, alkaline phosphatase, parathyroid hormone (PTH), and 1, 25-(OH)(2) vitamin D; and urine levels of calcium, phosphate, and creatinine were obtained. All measurements were made at study entry and at 2.0 kg of body weight. RESULTS: Despite a similar nutrient intake at advised levels for preterm infants, gains in body weight (g) and forearm bone length (cm), bone area (BA; cm(2)), bone mineral content (BMC; mg), and fat-free mass (g) were greater in PA infants. Forearm bone mineral density and fat mass gains did not differ between groups. Serum PICP levels remained constant in PA infants but decreased in C infants suggesting a slower rate of bone formation. Urine Pyd or bone resorption activity was similar between groups. A higher level of serum PTH was observed in PA infants at 2. 0 kg of body weight; however, the change from study entry to completion did not differ between groups. All other serum and urine values were similar and within normal limits. CONCLUSION: A daily PA program promotes greater gains in body weight, forearm length, BA, BMC, and fat-free mass in premature infants. PMID- 11061780 TI - Blood-brain phenylalanine relationships in persons with phenylketonuria. AB - OBJECTIVES: Clinicians caring for persons with phenylketonuria (PKU) have been perplexed by the occasional normal individual with the classical biochemical profile consistent with the diagnosis of PKU. Usually untreated subjects with the biochemical profile of blood phenylalanine (Phe) levels >1200 micromol/L are severely mentally retarded and may have neurological findings. Preliminary reports have recently appeared suggesting that low brain Phe levels, in comparison with elevated blood Phe levels, account for the occurrence of these occasional unaffected individuals with the biochemical profile consistent with PKU. METHOD: Magnetic resonance imaging/magnetic resonance spectroscopy was used to measure brain Phe content compared with simultaneously obtained blood Phe levels determined on the amino acid analyzer. This comparison was obtained in 5 normal non-PKU persons, 4 carriers of the gene causing PKU, and in 29 individuals with the proven form of the disorder. RESULTS: Blood-brain measurements in 5 normal persons ranged from.051 to.081 mmol/L, with a mean of.058 mmol/L. Their simultaneously measured brain levels of Phe ranged from.002 to.15 mmol/L, with a mean of.09 mmol/L. Similar measurements were obtained in 4 carriers of the gene causing PKU. Their blood levels varied between.068 and.109 mmol/L, with a mean of.091 mmol/L and simultaneously obtained brain levels of Phe varied between.06 and.21 mmol/L, with a mean of.11 mmol/L. Twenty subjects with a mean IQ of 104 exhibited a mean blood level of 1.428 mmol/L and a simultaneous mean brain level of.23 mmol/L, whereas 9 persons with a mean IQ of 98.7 exhibited a mean blood Phe level of 1.424 and a mean brain Phe level of.64 mmol/L. The correlation between blood and brain levels was not significant. CONCLUSION: In usual cases, intellectually normal persons who have never been treated but who have a biochemical profile consistent with classical PKU exhibit lower brain levels of Phe. Such individuals are exceptional and may not need the vigorous restriction of their blood Phe levels that is required by the majority of persons with PKU. PMID- 11061781 TI - Do parents understand immunizations? A national telephone survey. AB - BACKGROUND: Immunization may now be undervalued because vaccines have largely eliminated the threat of serious infectious diseases in childhood. As the incidence of vaccine-preventable diseases has declined, concern about vaccine safety has increased. Significant erosion of public confidence in vaccine safety could lead to reduced immunization rates and a resurgence of vaccine-preventable diseases. OBJECTIVE AND METHODS: To assess parents' understanding of vaccine preventable diseases, vaccines, immunization practices, and policies, we conducted a telephone survey in the United States with a nationally representative sample (n = 1600) of parents with children A (G71R), which is the most common mutation detected in the East Asian population, and the mutant enzyme had one third of the normal activity. G71R is the most common missense mutation we found in our analyses in Japanese patients with Gilbert's syndrome, and it corresponds to a UGT1A1 polymorphism in the Japanese population (the allele frequency is.16). One was heterozygous for 1456T-->G (Y486D) and homozygous for 211G-->A. Six were heterozygous for 211G-->A. One was heterozygous for both 211G-->A and a TATA box mutation (A(TA)7TAA). One had a heterozygous mutation in an enhancer region (C- >A at -1353). We did not detect a homozygous A(TA)7TAA mutation, which was the most common cause of Gilbert's syndrome in European population, in this study of Japanese infants with prolonged hyperbilirubinemia triggered by breast milk. CONCLUSIONS: The results indicate that defects of UGT1A1 are an underlying cause of the prolonged unconjugated hyperbilirubinemia associated with breast milk. One or more components in the milk may trigger the jaundice in infants who have such mutations. The mutations we found were identical to those detected in patients with Gilbert's syndrome, a risk factor of neonatal nonphysiologic hyperbilirubinemia and a genetic factor in fasting hyperbilirubinemia. PMID- 11061797 TI - Can urine clarity exclude the diagnosis of urinary tract infection? AB - OBJECTIVES: To determine the association of clear urine by visual inspection with the absence of significant bacteruria, and to compare it with standard urinalysis. METHODS: The study was performed in the emergency department of Children's Hospital Medical Center, Cincinnati, Ohio. It was a prospective, convenience sample of children <21 years of age who had catheterized or midstream clean-catch urine specimen collected for culture. Clinical findings including the presence or absence of fever, abdominal pain, dysuria, frequency, and urgency were collected for each patient. Urine was visually assessed for clarity by 2 independent observers using a standardized technique. Standard laboratory urinalysis and microscopy were also performed on all specimens. A positive urine culture was defined as >/=10(4) colony-forming unit (CFU)/mL of a urinary pathogen if obtained by catheterization and >/=10(5) CFU/mL if obtained by midstream. RESULTS: Samples were obtained from 159 patients ranging in age from 4 weeks to 19 years. Females comprised 77% of the patients. One hundred ten of the samples (69%) were clear to visual inspection. There were a total of 29 positive cultures; however, 3 were in children with clear urine. The finding of clear urine on visual inspection had a negative predictive value of 97.3%. These results were similar to those obtained with standard urinalysis. CONCLUSION: Clear urine on visual inspection cannot completely eliminate the possibility that a child has a urinary tract infection. However, it is a reproducible test that offers the advantages of being simple, fast, and inexpensive. The finding of clear urine should be considered a reasonable and relatively effective bedside screen for the presence of a urinary tract infection. PMID- 11061798 TI - Pneumococcal facial cellulitis in children. AB - OBJECTIVE: To review the epidemiology and clinical course of facial cellulitis attributable to Streptococcus pneumoniae in children. DESIGN: Cases were reviewed retrospectively at 8 children's hospitals in the United States for the period of September 1993 through December 1998. RESULTS: We identified 52 cases of pneumococcal facial cellulitis (45 periorbital and 7 buccal). Ninety-two percent of patients were <36 months old. Most were previously healthy; among the 6 with underlying disease were the only 2 patients with bilateral facial cellulitis. Fever (temperature: >/=100.5 degrees F) and leukocytosis (white blood cell count: >15 000/mm(3)) were noted at presentation in 78% and 82%, respectively. Two of 15 patients who underwent lumbar puncture had cerebrospinal fluid with mild pleocytosis, which was culture-negative. All patients had blood cultures positive for S pneumoniae. Serotypes 14 and 6B accounted for 53% and 27% of isolates, respectively. Overall, 16% and 4% were nonsusceptible to penicillin and ceftriaxone, respectively. Such isolates did not seem to cause disease that was either more severe or more refractory to therapy than that attributable to penicillin-susceptible isolates. Overall, the patients did well; one third were treated as outpatients. CONCLUSIONS: Pneumococcal facial cellulitis occurs primarily in young children (<36 months of age) who are at risk for pneumococcal bacteremia. They present with fever and leukocytosis. Response to therapy is generally good in those with disease attributable to penicillin-susceptible or nonsusceptible S pneumoniae. Ninety-six percent of the serotypes causing facial cellulitis in this series are included in the heptavalent-conjugated pneumococcal vaccine recently licensed in the United States. PMID- 11061799 TI - Day-to-day reactogenicity and the healthy vaccinee effect of measles-mumps rubella vaccination. AB - OBJECTIVE: Revaccination policies adopted in many countries to control measles have raised various safety issues including those concerning the second vaccine dose. We performed a prospective, double-blind, crossover trial among twins receiving a measles-mumps-rubella (MMR) vaccine. STUDY DESIGN: The study comprised 1162 monozygous and heterozygous twins, each of whom randomly received placebo and then vaccine, or vice versa, 3 weeks apart, at 14 to 83 months of age. Most of the oldest children had previously been vaccinated against measles, and one half of the remainder of children had had the disease. Symptoms and signs were recorded daily on structured forms. Statistical methods included a complex analysis of the vaccine attributability of the symptoms and conditional logistic regression. RESULTS: Vaccination-attributable events occurred in 6% overall. At 14 to 18 months of age, reactions developed between days 6 and 14, peaking at day 10. The clearest vaccine-attributable effect was fever exceeding 101.3 degrees F (38. 5 degrees C; odds ratio: 3.28; 95% confidence interval: 2.23-4.82; P <.001), but the same trend was found for rash, arthralgia, conjunctivitis, staying in bed, drowsiness, and irritability. At 6 years of age, systemic reactions occurred 5 to 15 times less frequently, only arthralgia being associated with vaccination. Zygocity, gender, history of allergy, or infections did not modify reactions. Instead, respiratory symptoms developed within days postinjection to a level of 15% to 20% without subsequent decline and with no difference between vaccinees and placebo recipients. CONCLUSION: Vaccination was avoided during infections, but many small children became mildly ill within a week or so with no relation to vaccination (the healthy vaccinee effect). MMR vaccine was virtually nonreactogenic when given at 6 years of age. vaccine, measles, mumps, rubella, reactogenicity, adverse events, zygocity, healthy vaccinee effect. PMID- 11061800 TI - Should central venous catheters be removed as soon as candidemia is detected in neonates? AB - BACKGROUND: Controversy exists regarding the most appropriate acute management of central venous catheters (CVCs) in neonates with candidemia, with up to two thirds of neonatologists preferring to attempt antifungal therapy without removing CVCs. OBJECTIVE: To determine whether CVCs should be removed as soon as candidemia is detected in neonates. Methods. A cohort study of candidemia and CVC was conducted in infants in a neonatal intensive care unit (NICU) over a 5-year period (1994-1998). RESULTS: Fifty infants had early-removal CVC (ER-CVC) within 3 days and 54 infants had late-removal CVC (LR-CVC) >3 days after the first positive blood culture for Candida species. All infants were treated with amphotericin B. There was no significant difference between infants in the ER-CVC and LR-CVC groups in terms of gender, ethnicity, birth weight, gestational age, age at candidemia, severity-of-illness scores, distribution of types of CVC, or in the distribution of Candida species causing candidemia. The ER-CVC group had significantly shorter duration of candidemia (median: 3 days; range: 1-14 days), compared with the LR-CVC group (median: 6 days; range: 1-24 days). The case fatality rate of Candida albicans candidemia was significantly affected by the timing of CVC removal: 0 of 21 (95% confidence interval [CI]: 0-14) infants died in the ER-CVC group in contrast to 9 of 23 (39%; 95% CI: 19-59) in the LR-CVC group. CONCLUSION: Failure to remove CVC as soon as candidemia was detected in neonates was associated with significantly increased mortality in C albicans candidemia and prolonged duration of candidemia regardless of Candida species. PMID- 11061801 TI - Snoring in Portuguese primary school children. AB - OBJECTIVE: To determine the prevalence of snoring and its potential associations with sleep problems, such as daytime symptoms, medical conditions, school performance, and behavioral disturbances in Portuguese children attending primary school. METHODS: A previously validated questionnaire was sent to the parents of 1381 children attending primary schools in a parish of Coimbra, Portugal. To assess behavioral disturbances, the Portuguese version of Rutter's Children's Behavior Questionnaire for completion by teachers was used. RESULTS: Of the 988 questionnaires returned (71.5%), complete information concerning snoring was obtained for 976 children (496 girls and 480 boys; mean age: 8.1 +/- 1.5 years). Loud snoring during sleep was reported as frequent or constantly present (LSn) in 84 children (8.6%), as occasionally present in 299 children (30.6%), and as never present (NSn) by 593 children (60.8%). The LSn and NSn groups did not differ with respect to age, gender, sleep duration, time to fall asleep, frequency of night wakings, bedwetting, daytime tiredness, and school achievement. However, LSn was significantly associated with increased bedtime problems (fears and struggles), increased need for comforting activities to fall asleep, behaviors suggestive of parasomnias (sleep talking, teeth grinding, and night terrors), increased daytime sleepiness and irritability, and behavioral disturbances. Children in the LSn group were also more likely to report recurrent medical problems particularly those involving infections of the respiratory tract. CONCLUSIONS: Snoring is a common symptom in Portuguese children that is associated with behavioral daytime and sleep time disturbances. Children with loud snoring may benefit from early evaluation and intervention. PMID- 11061802 TI - Ocular injuries by elastic cords in children. AB - BACKGROUND: Elastic cords hitting the eyeball as high-speed projectiles can severely damage ocular structures and can produce permanent visual function impairment. OBJECTIVES: To evaluate the frequency, mechanics, and severity of eye injuries caused by elastic cords in children to adopt the most appropriate preventive measures. METHODS: A retrospective medical records review of hospital admissions secondary to ocular trauma between 1991 and 1997 in a pediatric ophthalmology unit at an urban tertiary care pediatric hospital was performed to select all children admitted for ocular injury caused by an elastic cord. RESULTS: Eight children fulfilled the inclusion criteria; the prevalence ratio was 2% of all pediatric trauma admissions. In all cases the mechanics of trauma was a combination of blunt and high-speed projectile injury. The mechanism of trauma in younger patients was typically a cord that was misused during unsupervised playtime, whereas cord slipping from car roof racks was noted in older patients. One patient suffered a severe permanent visual impairment caused by retinal detachment. All other children regained full visual acuity at the time of discharge and maintained it through a mean follow-up of 22 months (range: 18 29). CONCLUSION: Circumstances of injury in younger children are different from those found in older children, the latter being similar to those reported for adults. Prevention is the primary measure to be taken to reduce the prevalence of this injury and to lower the risk for ocular severe anatomic damage as much as possible. This can be achieved primarily by modifying the design of the hooks, intensifying educational campaigns, and keeping elastic cords out of children's reach. PMID- 11061803 TI - The pediatrician's role in reducing tobacco exposure in children. AB - OBJECTIVE: Pediatricians have a unique and important role to play in the prevention and treatment of childhood and adolescent tobacco use, the protection of patients from the harmful effects of environmental tobacco smoke, and the encouragement of smoking cessation among parents. However, because recent research indicates that physician training in tobacco dependence is woefully weak and lacks a model for training, this article constructs a useful approach to this problem. METHODOLOGY: A comprehensive review of the literature served as the basis for the development of a new model for pediatrician training in tobacco dependence. RESULTS: A comprehensive model is presented for training pediatricians in the areas of reducing infant and child exposure to environmental tobacco smoke, preventing youth smoking initiation, and providing smoking cessation assistance for adolescents and parents. CONCLUSIONS: Pediatricians have been called on to play an active role in the antitobacco arena. Because of their unique opportunity to interact with children, adolescents, and parents, pediatricians can and should be antitobacco interventionists. For this to occur, however, additional guidance should be provided to pediatricians during their training to better prepare them to carry out effective assessment and intervention practices. smoking initiation, smoking prevention, smoking cessation, environmental tobacco smoke, pediatricians. PMID- 11061804 TI - Major factors influencing breastfeeding rates: Mother's perception of father's attitude and milk supply. AB - OBJECTIVE: To determine factors influencing feeding decisions, breastfeeding and/or bottle initiation rates, as well as breastfeeding duration. SETTING: A family medicine practice of a 530-bed community-based hospital in northwestern Pennsylvania. PARTICIPANTS: All mothers whose infants received well-child care from birth to 1 year of age. OUTCOME MEASURE: A survey of 28 simple questions was developed and mailed to 245 mothers. The survey assessed: 1) demographics, 2) prenatal and postnatal care, 3) sources of breastfeeding information, 4) timing of decision, 5) preference, 6) type of feeding selected, 7) duration of breastfeeding, 8) factors influencing decisions to breastfeed and/or to bottle feed, and 9) factors that would have encouraged bottle-feeding mothers to breastfeed. RESULTS: The breastfeeding initiation rate was 44.3%. By the time the infant was 6 months old, only 13% of these were still breastfeeding. The decision to breastfeed or to bottle-feed was most often made before pregnancy or during the first trimester. The most common reasons mothers chose breastfeeding included: 1) benefits the infant's health, 2) naturalness, and 3) emotional bonding with the infant. The most common reasons bottle-feeding was chosen included: 1) mother's perception of father's attitude, 2) uncertainty regarding the quantity of breast milk, and 3) return to work. By self-report, factors that would have encouraged bottle-feeding mothers to breastfeed included: 1) more information in prenatal class; 2) more information from TV, magazines, and books; and 3) family support. CONCLUSIONS: To overcome obstacles, issues surrounding perceived barriers, such as father's attitude, quantity of milk, and time constraints, need to be discussed with each parent. To achieve the goal of 75% of breastfeeding mothers, extensive education regarding the benefits must be provided for both parents and optimally the grandmother by physicians, nurses, and the media before pregnancy or within the first trimester. PMID- 11061805 TI - Distribution of accidents, injuries, and illnesses by family type. ALSPAC Study Team. Avon Longitudinal Study of Pregnancy and Childhood. AB - OBJECTIVE: To investigate whether family type and psychosocial risks indexed by family type were systematically associated with differences in health outcomes in children. DESIGN AND SUBJECTS: The study is based on a longitudinal, prospective study of a large (n = approximately 10 000) community sample of families, the Avon Longitudinal Study of Pregnancy and Childhood. MAIN OUTCOME MEASURES: Frequency of accidents, illnesses, and medical interventions. RESULTS: At 2 years of age, children in single-parent and stepfamilies were disproportionately likely to experience accidents and receive medical treatment for physical illnesses. In addition, children in single-parent families and stepfamilies were more likely to be hospitalized or receive attention from a hospital doctor for an injury or illness. Exposure to psychosocial risks also were elevated in single-parent families and stepfamilies, compared with intact or nonstepfamilies, and these factors primarily accounted for the connection between family type and children's physical health. CONCLUSIONS: The consequences of family transitions on children's health extend beyond traditional mental health and behavioral outcomes and include accident proneness, illness, and receipt of medical attention. The mediating processes are not entirely attributable to social class differences connected to family type and may instead be associated with a range of psychosocial risks that are more frequently found in single-parent families and stepfamilies, compared with intact or nonstepfamilies. Prevention and intervention efforts directed toward children at risk for poor behavioral and mental health adjustment secondary to family disruption should consider children's physical health and health-related behaviors. PMID- 11061806 TI - Targeting lead screening: The Ohio Lead Risk Score. AB - OBJECTIVE: Annual blood lead (BPb) screening is recommended for children /=12% of BPb test results >/=10 microg/dL. RESULTS: Data from 897 census tracts were available. Higher risk for lead toxicity existed in areas where: 1) >/=55% of houses were built before 1950 (adjusted odds ratio [AOR]: 10.9 [6.1,19.6]); 2) >/=35% of residents were black (AOR: 3.5 [2.0,6. 3]); 3) >/=35% of residents had less than a high school education (AOR: 6.1 [3.6,10.4]); and 4) >/=50% of housing units were renter-occupied (AOR: 3.6 [2.1,6.2]). Receiver operator characteristic (ROC) curves demonstrated no significant differences after applying the model in a second dataset. CONCLUSIONS: Several community characteristics predict risk for lead toxicity in children and may provide a useful approach to focus lead screening, especially in communities where public health resources are limited. The approach described here may also prove helpful in identifying factors within a community associated with other environmental public health hazards for children. PMID- 11061807 TI - Sleep and periodic limb movement in sleep in juvenile fibromyalgia. AB - OBJECTIVES: Fibromyalgia has been recently recognized in children and adolescents as juvenile fibromyalgia (JF). In adult fibromyalgia, subjective complaints of nonrestorative sleep and fatigue are supported by altered polysomnographic findings including a primary sleep disorder known as periodic limb movements in sleep (PLMS) in some subjects. Although poor sleep is a diagnostic criterion for JF, few reports in the literature have evaluated specific sleep disturbances. Our objectives were to evaluate in a controlled study the polysomnographic findings of children and adolescents with JF for alterations in sleep architecture as well as possible PLMS not previously noted in this age group. METHODS: Sixteen consecutive children and adolescents (15.0 +/- 2.6 years of age) diagnosed with JF underwent overnight polysomnography. Polysomnography was also performed on 14 controls (14.0 +/- 2.2 years of age) with no history of an underlying medical condition that could impact on sleep architecture. Respiratory variables, sleep stages, and limb movements were measured during sleep in all subjects. RESULTS: JF subjects differed significantly from controls in sleep architecture. JF subjects presented with prolonged sleep latency, shortened total sleep time, decreased sleep efficiency, and increased wakefulness during sleep. In addition, JF subjects exhibited excessive movement activity during sleep. Six of the JF subjects (38%) were noted to have an abnormally elevated PLMS index (>5/hour), indicating PLMS in these subjects. CONCLUSION: Our study demonstrated abnormalities in sleep architecture in children with JF. We also noted PLMS in a significant number of subjects. This has not been reported previously in children with this disorder. We recommend that children who are evaluated for JF undergo polysomnography including PLMS assessment. juvenile fibromyalgia; periodic limb movement in sleep; restless legs syndrome. PMID- 11061808 TI - Chest compressions in an infant with osteogenesis imperfecta type II: No new rib fractures. AB - The case report of a newborn female with osteogenesis imperfecta type II who underwent cardiopulmonary resuscitation (CPR) with manual chest compressions for several minutes is presented. Chest radiographs taken before and after the chest compressions were administered were reviewed by several radiologists from 3 different hospitals and demonstrated no new radiographically visible rib fractures. Collagen analysis, the patient's clinical appearance, and clinical course, as well as a consultant's opinion aided in confirmation of the diagnosis of osteogenesis imperfecta type II. A review of 4 previous studies concerning rib fractures and CPR is included. This unique case supports previous articles that have concluded that rib fractures rarely, if ever, result from CPR in pediatrics, even in children with a lethal underlying bone disease, such as osteogenesis imperfecta type II. cardiopulmonary resuscitation, chest compressions, osteogenesis imperfecta, rib fractures, bone disease. PMID- 11061809 TI - Early puberty: rapid progression and reduced final height in girls with low birth weight. AB - OBJECTIVE: To assess whether, in girls with early onset of puberty, low birth weight is a risk factor for rapid progression to menarche and for short adult stature. DESIGN: Longitudinal clinical assessment of 54 Catalan (Northern Spanish) girls followed from early onset of puberty (onset of breast development between 8.0 and 9.0 years of age) to final height. The timing of menarche and the final height were analyzed a posteriori according to birth weight, the cutoff level between normal and low birth weight subgroups being -1.5 standard deviation (SD; approximately 2.7 kg at term birth). RESULTS: Normal and low birth weight girls had similar target heights and characteristics at diagnosis of early puberty. However, menarche occurred on average 1.6 years earlier in low versus normal birth weight girls (11.3 +/-.3 years vs 12.9 +/-.2 years), and final height was >5 cm shorter in low birth weight girls (parental adjusted height SD: .6 +/-.2 cm vs.3 +/-.2 cm). CONCLUSION: The timing of menarche and the level of final height in Catalan girls with early onset of puberty was found to depend on prenatal growth. Girls with normal birth weight tend to progress slowly through puberty with a normal timing of menarche and normal final height. In contrast, girls with low birth weight tend to progress relatively rapidly to an early menarche and to a reduced final height. If these findings are confirmed in other ethnic and/or larger groups, then a subgroup has been identified that will most likely benefit from any therapeutic intervention aiming at a delay of pubertal development and/or an increase of final height. PMID- 11061810 TI - Growth pattern of breastfed and nonbreastfed infants with atopic dermatitis in the first year of life. AB - OBJECTIVE: The growth of infants with atopic dermatitis (AD) has been poorly investigated based on the early type of feeding. The aim of this study was to assess the growth pattern of AD infants during the first 12 months of life in comparison to healthy infants, according to the early type of feeding (breastfed or nonbreastfed). METHODS: Fifty-five term AD infants (36 breastfed and 19 nonbreastfed) and 114 term healthy infants (58 breastfed and 56 nonbreastfed) were evaluated by standardized growth indices (z scores; National Center for Health Statistics-World Health Organization data) through the first 12 months of life. RESULTS: No difference was found between AD and healthy groups at birth. In AD infants, weight (WA) and length (LA) z scores decreased with age and were significantly lower, compared with healthy infants from the second month of age onward. The difference of mean z scores between AD and healthy infants at 12 months of age was -.69 (95% confidence interval [CI]: -1.00 to -.38) for WA and .67 (95% CI: -.98 to -.36) for LA. The growth pattern of AD infants was not influenced by the early type of feeding, whereas in the 6- to 12-month period, the delay in growth was more pronounced in patients with more severe dermatitis. CONCLUSIONS: In the first year of life, AD infants show a progressive impairment in growth irrespective of the early type of feeding. The severity of disease may be an independent factor negatively influencing growth. PMID- 11061816 TI - Research priorities in complementary feeding: International Paediatric Association (IPA) and European Society of Paediatric Gastroenterology, Hepatology, and Nutrition (ESPGHAN) workshop. PMID- 11061817 TI - Canadian recommendations. PMID- 11061818 TI - Patterns of complementary feeding (weaning) in countries of the European Union: topics for research. PMID- 11061819 TI - American Academy of Pediatrics recommendations for complementary feeding. PMID- 11061820 TI - Overview of complementary feeding (weaning) in countries of Asia. PMID- 11061821 TI - Feeding practices, growth, and morbidity in Tunisia. PMID- 11061822 TI - Infant feeding and weaning practices in an urbanizing traditional, hunter gatherer society. PMID- 11061823 TI - Taste acquisition and appetite control. PMID- 11061824 TI - Taste and appetite in infancy: possible topics for research. PMID- 11061825 TI - Complementary feeding and neuromuscular development. PMID- 11061826 TI - Complementary feeding and genetic programming of gastrointestinal function. PMID- 11061827 TI - Gastrointestinal digestive and absorptive function. PMID- 11061828 TI - Complementary feeding and infant growth and body composition. PMID- 11061829 TI - The influence of breastfeeding and complementary foods on growth until three years of age in the Euro-Growth Study. PMID- 11061830 TI - Feeding mode, infections, and anthropometric status in early childhood. PMID- 11061831 TI - Potential renal solute load: considerations relating to complementary feedings of breastfed infants. PMID- 11061832 TI - Complementary foods and the development of food allergy. PMID- 11061833 TI - Complementary feeding and enteropathies. PMID- 11061834 TI - Complementary feeding and later health. PMID- 11061835 TI - Determining limiting nutrients by linear programming: A new approach to predict insufficient intakes from complementary foods. PMID- 11061836 TI - WHO/UNICEF review on complementary feeding and suggestions for future research: WHO/UNICEF guidelines on complementary feeding. PMID- 11061837 TI - Complex carbohydrates and sugars. PMID- 11061838 TI - Protein and amino acids. PMID- 11061839 TI - Are there negative effects of an excessive protein intake? PMID- 11061840 TI - Lipids in complementary foods. PMID- 11061841 TI - Iron and zinc intake from complementary foods: some issues from Pakistan. PMID- 11061842 TI - The adequacy of micronutrients in complementary foods. PMID- 11061843 TI - Improving complementary feeding practices and responsive parenting as a primary component of interventions to prevent malnutrition in infancy and early childhood. PMID- 11061844 TI - Complementary feeding and breastfeeding. PMID- 11061845 TI - Cows' milk in complementary feeding. PMID- 11061846 TI - Research priorities on safety of complementary feeding. PMID- 11061847 TI - Paving with good intentions: learning from HCV lookback. PMID- 11061848 TI - NAT and blood safety: what is the paradigm? PMID- 11061849 TI - Why is there such difficulty in defining the natural history of hepatitis C? PMID- 11061850 TI - NAT of the United States and Canadian blood supply. PMID- 11061851 TI - An evaluation of the process and costs associated with targeted lookbacks for HCV and general notification of transfusion recipients. AB - BACKGROUND: The Commission of Inquiry on the Blood System in Canada recommended that hospitals notify patients who received blood between 1978 and May 1990 of the risks of contracting HIV (up to the end of 1985 only) and HCV infection. The commission also recommended that patients should be informed of any transfusion received. STUDY DESIGN AND METHOD: General notifications for HIV and HCV for this period were begun in mid-1994. Notification after discharge of transfusions received after May 1990 was begun in 1997. Targeted HCV lookback was performed from 1995 to 1999. RESULTS: Of 21,016 transfusion recipients from January 1978 to May 1990 identified in the general look-back process and believed still alive, 13,549 (64%) were presumed contacted, by registered mail. The overall contact rate for the ongoing notifications (transfusions after May 1990) cannot be accurately determined, as registered mail was not used and a reply not requested. The total cost for these two processes was CAN$373,481, or $13 per patient believed contacted. Most (56%) of this cost was for the conversion to electronic form of paper transfusion records for the period 1978 through early 1984. In the targeted HCV lookback program 1995 through 1999, 94 percent of 256 recipients of specific components identified as likely to have transmitted HCV either were contacted or had died. Of 84 living recipients, 47 (56%) are HCV positive. The last documented potential seroconversion occurred after a transfusion in November 1991, during the period of first-generation EIA testing. If the targeted HCV lookback had been restricted to transfusions after 1987, as the FDA recommended, we would have failed to identify 39 living patients, of whom 21 are HCV positive. The cost per HCV-positive patient notified in the targeted HCV lookback was CAN $4,174. CONCLUSION: The cost of compliance with the com-mission's recommendations was CAN$569,636. Over 28,000 of 36,773 transfusion recipients were notified or presumed notified, and 272 targeted HCV lookbacks to 256 recipients were performed. Performance of this task required the existence of transfusion records back to 1978, conversion of paper records to electronic form, and adequate secretarial and financial support. PMID- 11061852 TI - Evaluation of the effectiveness of targeted lookback for HCV infection in the United States-interim results. AB - BACKGROUND: As part of a nationwide program to identify persons at increased risk for HCV infection, persons who received blood from donors who later tested positive for anti-HCV are being directly notified. STUDY DESIGN AND METHODS: In December 1999, all 198 blood collection establishments (BCEs) and 5442 hospital transfusion services (TSs) in the United States were surveyed by mailed questionnaire to evaluate their progress in carrying out this notification. RESULTS: Eighty-one percent of the BCEs and 64 percent of the TSs responded. After correcting for nonresponse, an estimated 98,484 components at potential risk for transmitting HCV, according to previous testing of multiantigen-screened donors, were identified nationwide, of which 85 percent had been transfused to recipients. Lookback for these recipients was completed for 80 percent, of whom 69 percent had died. Of those living, 78 percent were successfully notified. An estimated 49.5 percent of those notified were tested; 18.9 percent of those tested were anti-HCV positive, and 32 percent of that group knew they were positive before notification. On the basis of an 85.5 percent reported completion rate for component notifications back through 1988, an estimated 1520 persons will have been newly identified as anti-HCV-positive when lookback related to multiantigen screening of donors is completed. CONCLUSION: Targeted lookback related to previous multiantigen screening of donors will identify <1 percent of the estimated 300,000 HCV-positive persons in the United States who may have acquired their infection via blood transfusion. PMID- 11061853 TI - A model of the health and economic impact of posttransfusion hepatitis C: application to cost-effectiveness analysis of further expansion of HCV screening protocols. AB - BACKGROUND: Cost-effectiveness analyses are needed to decide the value of further expansion of the screening protocols for HCV in blood donors. However, such analyses are hampered by imperfect knowledge of the health and economic repercussions of posttransfusion hepatitis C (PTHC). STUDY DESIGN AND METHODS: A Monte Carlo simulation of a Markov model representing the outcomes of patients transfused with HCV-infective blood was used to estimate the health and economic impact of PTHC and to calculate the cost-effectiveness ratio of various HCV screening methods. RESULTS: Median survival for hypothetical patients with PTHC and for controls without hepatitis was 11.25 and 11.75 years, respectively. Overall, 12.3 percent of patients receiving HCV-infective blood will develop chronic hepatitis, 9.3 percent will progress to liver failure, and 9. 25 percent will eventually die of liver disease after a median time of 20.75 years (range, 6 70). Ninety-one percent of the infected blood recipients had no reduction in life expectancy due to PTHC, and the average loss per patient was 0.754 years. The present value of the lifetime health costs incurred by patients with PTHC is $6330 per case. HCV antibody testing increases the patients' life expectancy by 20.4 hours per blood collection tested, and it results in net savings by decreasing the number of patients that will require treatment for liver disease in the future. Adding HCV NAT increases the patients' life expectancy by 0.08 hours per blood collection tested, at a cost-effectiveness ratio of $1,829,611 per QALY gained. CONCLUSION: PTHC has low health benefits because of the advanced age of many blood recipients. Testing donors for HCV antibodies results in net savings for the health care system, despite low health benefits. Adding HCV NAT would produce little additional gain at a very high cost. PMID- 11061854 TI - Testing of individual blood donations for HCV RNA reduces the residual risk of transfusion-transmitted HCV infection. AB - BACKGROUND: To allow cost-effective RNA testing with NAT techniques, the national authorities of several countries have planned or already introduced tests of mixed specimens, that is, plasma pools. STUDY DESIGN AND METHODS: High-throughput extraction, amplification, and detection of HCV RNA from individual blood donations were optimized and validated. The feasibility of the method and the frequency of anti-HCV-negative, HCV RNA-positive donations were determined in a prospective study of 27,745 allogeneic and 792 autologous individual donations. RESULTS: The 50- and 95-percent detection limits of the method were determined at 44 IU per mL and 162 IU per mL, respectively (World Health Organization HCV reference material). When 201 HCV RNA-positive sera were taken as a reference, the sensitivity was 97.5 percent. The assay specificity was determined at 99.77 percent. During a 20-month period, two seronegative blood donors tested positive in HCV PCR. The viral load of these donations was 6 x 10(6) and 3 x 10(7) copies per mL, respectively. Thus, the yield of HCV RNA testing in this study was 7. 63 per 100,000 screened donations (95% CI, 1.25-22.07). In both PCR-positive donors, seroconversion was found in subsequent blood samples. CONCLUSION: This study compares the feasibility of single-donation HCV RNA screening, with the detection of a relatively high percentage of window-phase donations, to data reported from groups using HCV RNA testing of plasma pools. The relative yield of NAT of individual donations versus minipools should be directly investigated in the near future. PMID- 11061855 TI - Efficacy of HCV core antigen detection during the preseroconversion period. AB - BACKGROUND: The purpose of this study was to compare the performances of HCV core antigen (HCV Ag) testing with HCV RNA detection during the preseroconversion period. STUDY DESIGN AND METHODS: Six HCV antibody (HCV Ab)-negative and HCV RNA positive blood samples from 6 donors and 135 serial samples from 28 patients who had undergone hemodialysis, collected a mean of 90 days before the detection of HCV Ab, were tested by ELISA for the detection of HCV Ag and by PCR to quantify HCV RNA. RESULTS: Five of the six donors were positive for HCV Ag. The donor with a negative HCV Ag test had the lowest viral load. In the hemodialysis patients, the 43 first specimens of the series were HCV RNA negative. Of the 92 specimens that were HCV RNA positive, 81 (88%) were positive for HCV Ag. Among the 74 samples with more than 10(5) RNA copies, 71 (96%) were HCV Ag positive. Average time from first viremic bleed to first HCV Ag-positive bleed was estimated at 2.0 days and that to first HCV Ab-positive bleed at 50.8 days. CONCLUSION: HCV Ag testing permits the detection of an HCV infection about 1.5 months earlier than the HCV Ab screening tests and an average of only 2 days later than quantitative HCV RNA detection in individual specimens. PMID- 11061856 TI - Clinical illness due to parvovirus B19 infection after infusion of solvent/detergent-treated pooled plasma. AB - BACKGROUND: Lipid-enveloped viruses such as HIV, HBV, and HCV can be inactivated by treatment with solvents and detergents. HAV and human parvovirus B19 lack lipid envelopes and are not inactivated. Solvent/detergent-treated pooled plasma (S/D plasma) contains neutralizing antibodies, but it is not known whether the parvovirus B19 antibody content is sufficient to prevent transmission of the disease. A patient is described who developed a clinical illness due to parvovirus B19 infection after the infusion of S/D plasma. CASE REPORT: A 36-year old woman with myasthenia gravis underwent five plasma exchange procedures from January 15 to January 25, 1999, using albumin, except for 5 units of SD plasma given because of a low fibrinogen level. Four of the 5 units were implicated in a recall after high levels of parvovirus B19 DNA were found in several lots. Two weeks after the infusion, the patient developed fatigue, a rash, and severe polyarthralgias. Parvovirus B19 IgG and IgM antibody titers were consistent with an acute infection. CONCLUSION: Clinically apparent parvovirus B19 infection can follow the use of S/D plasma that contains high levels of parvovirus B19 DNA. PMID- 11061857 TI - Transfusion errors in New York State: an analysis of 10 years' experience. AB - BACKGROUND: While public focus is on the risk of infectious disease from the blood supply, transfusion errors also contribute significantly to adverse outcomes. This study characterizes such errors. STUDY DESIGN AND METHODS: The New York State Department of Health mandates the reporting of transfusion errors by the approximately 256 transfusion services licensed to operate in the state. Each incident from 1990 through 1998 that resulted in administration of blood to other than the intended patient or the issuance of blood of incorrect ABO or Rh group for transfusion was analyzed. RESULTS: Erroneous administration was observed for 1 of 19, 000 RBC units administered. Half of these events occurred outside the blood bank (administration to the wrong recipient, 38%; phlebotomy errors, 13%). Isolated blood bank errors, including testing of the wrong specimen, transcription errors, and issuance of the wrong unit, were responsible for 29 percent of events. Many events (15%) involved multiple errors; the most common was failure to detect at the bedside that an incorrect unit had been issued. CONCLUSION: Transfusion error continues to be a significant risk. Most errors result from human actions and thus may be preventable. The majority of events occur outside the blood bank, which suggests that hospitalwide efforts at prevention may be required. PMID- 11061858 TI - Evaluation of an automated system for the collection of packed RBCs, platelets, and plasma. AB - BACKGROUND: This study evaluated the quality of WBC-reduced platelets, RBCs, and plasma collected on a new system (Trima, Gambro BCT) designed to automate the collection of all blood components. The study also evaluated donor safety and suitability of these components for transfusion. STUDY DESIGN AND METHODS: In Phase I, the quality of the components collected on the new system was evaluated by standard in vitro and in vivo testing methods. Results were compared to those from control components collected by currently approved standard methods. In Phase II, additional collections were performed to evaluate the acceptability of the new system and the safety of platelets collected. RESULTS: In vivo 24-hour RBC recovery was 76.8 +/- 3.1 percent for the test RBC units and 77.1 +/- 4.4 percent recovery for whole-blood (control) RBCs. The differences between test and control platelet results in the in vivo and in vitro assays were not clinically significant. Plasma clotting factors and fibrinogen levels met international standards. The system was well accepted by donors, and no major adverse donor reactions were reported for the 68 procedures performed. No problems were reported with transfusing the blood components collected. CONCLUSION: Blood components collected with the Trima are equivalent to currently available components, and they meet the applicable regulatory standards. This system provides consistent, standardized components with predictable yields. It provides the option of fully automating the collection of all blood components. PMID- 11061859 TI - Leukapheresis components may be cryopreserved at high cell concentrations without additional loss of HPC function. AB - BACKGROUND: The aim of this study was to assess the feasibility of freezing mobilized peripheral blood progenitor cell (PBPC) components at higher cell concentrations than are classically recommended for bone marrow. This approach might have potential benefits, such as lower cost of processing and storage and less risk of the complications associated with the transfusion of large component volumes and large quantities of DMSO. STUDY DESIGN AND METHODS: In the first phase, small aliquots of 19 apheresis components were cryopreserved at standard and higher cell concentrations (Aliquots A and B, respectively). In the second phase, 21 apheresis components were split into two bags each and frozen at standard (Bag A) and high (Bag B) cell concentrations. The differences in viability, cloning efficiency, and nucleated cell recovery in Bags A and B were examined. Finally, the hematologic recovery of 10 patients who underwent autologous transplantation with PBPC components frozen at high cell concentrations was analyzed. RESULTS: The median cell concentration at freezing was 94 (57-100) x 10(6) per mL and 291 (220-467) x 10(6) per mL for Aliquots A and B, respectively, and 90.9 (45.4-92) x 10(6) per mL and 332 (171-582) x 10(6) per mL for Bags A and B, respectively. The viability was significantly lower in samples frozen at higher cell concentrations: 92 versus 83 percent (p = 0.001) and 87 versus 77 percent (p<0.001) for Aliquots and Bags A and B, respectively. Significant differences were not observed in the recovery of total nucleated cells (102 vs. 101% and 98 vs. 105%) or the cloning efficiency after thawing (13 vs. 16% and 27 vs. 23%) for Aliquots and Bags A and B, respectively. The time to granulocyte engraftment >0.5 x 10(9) per L and platelet engraftment >20 x 10(9) per L was 9 (8-11) and 10.5 (7-21) days, respectively. CONCLUSION: The cryopreservation of PBPC components at standard concentrations and 3.3 (1.8-6.2) fold cell concentrations has no adverse effect on the function of HPCs after thawing. PMID- 11061860 TI - Preoperative predictors of the need for allogeneic blood transfusion in lung cancer surgery. AB - BACKGROUND: The use of blood-saving techniques in elective surgery can produce a favorable cost-benefit ratio only when there is a reasonable likelihood that transfusion will be required. To apply a targeted blood-sparing technique in lung cancer surgery, the patient's preoperative characteristics that predict the use of allogeneic blood transfusion (ABT) in this practice were investigated. STUDY DESIGN AND METHODS: One hundred seventy-three consecutive patients who underwent primary lung cancer surgery were included in this retrospective study. Clinical and epidemiologic variables, lung tumor extension (TNM staging), and surgery type were analyzed by logistic regression to discover the preoperative predictors of ABT. RESULTS: Thirty patients, 17.3 percent of all who underwent surgery and 19.9 percent of those who underwent resolvent surgery, received ABT. Excluding a patient who needed 18 units of RBCs, the number of ABT units required by transfused patients was 1. 93 +/- 0.88 (mean +/- SD). Extensive surgery, patient's age (< or =64 years), and elevated erythrocyte sedimentation rate (>45 mm/hour) were the preoperative variables that influenced the need for ABT. The definitive predictive model was able to recognize 82.3 percent of patients who received ABT and 95.6 percent of those who did not. CONCLUSION: A predictive model can preoperatively identify patients at risk for needing ABT in lung cancer surgery. The model could be utilized to tailor blood-sparing intervention programs. PMID- 11061861 TI - Total hip replacement surgery does not influence RBC survival. AB - BACKGROUND: RBC survival may be affected by mechanical or oxidative stress as well as the inflammatory effect of surgery, and thus may contribute to postoperative anemia. STUDY DESIGN AND METHODS: RBC survival was studied in otherwise healthy patients who underwent total hip replacement (THR). RBCs of five patients scheduled for elective THR surgery were labeled with (51)Cr 2 weeks before the surgical procedure. The disappearance of (51)Cr-labeled RBCs was calculated for the preoperative period and for the period from postoperative Day 4 on, when blood loss had ceased. RESULTS: The half-life of (51)Cr-labeled RBCs did not change if estimated before (29.0 +/- 4.4 days) and after (27.4 +/- 3.6 days, p = 0.55) surgery. This indicates that the lifespan of RBCs is not influenced by the surgical procedure. CONCLUSION: A decrease in RBC survival does not contribute to the pathogenesis of the anemia found after THR surgery. PMID- 11061862 TI - Quantitative determination of anti-K (KEL1) IgG and IgG subclasses in the serum of severely alloimmunized pregnant women by ELISA. AB - BACKGROUND: Severe cases of HDN occur after the immunization of the mother with K (KEL1) antigen. To date, the only means of evaluating the concentration of anti-K in maternal serum is by titration with an indirect antiglobulin test (IAT). A more accurate estimation of the serum anti-K concentration is needed. STUDY DESIGN AND METHODS: An ELISA technique was developed for the determination of the absolute concentration of anti-K IgG and IgG subclasses in the sera of alloimmunized patients. In this technique, after absorption of anti-K on K positive RBCs and subsequent elution at acid pH, the concentration of anti-K in the eluate was measured with a sensitive and reproducible ELISA. This method was validated with monoclonal and polyclonal anti-K. It was then used to assay the sera of eight pregnant women with anti-K immunization, associated with early fetal anemia (Hct, 7-17%) detected between the 20th and the 31st week of pregnancy. In addition, in most of these cases, the anemia was associated with fetal hydrops. RESULTS: The anti-K IgG concentration measured by ELISA in the sera of the eight women varied from 1.0 to 4.1 microg per mL (mean, 2.2 microg/mL). Therefore, severe and early forms of fetal anemia can be observed with a relatively low concentration of anti-K (as compared to the concentration of anti-D in similar cases of fetal anemia due to anti-D). The mean proportion of each IgG subclass of anti-K in these sera was IgG1, 95.9 percent; IgG2, 2.4 percent; IgG3, 1.3 percent; and IgG4, 0.4 percent. CONCLUSION: A simple method for quantitative estimation of anti-K in human serum has been developed. Low concentrations of anti-K can cause fetal anemia relatively early in pregnancy. This method should lead to a better identification of pregnant women whose fetuses are at risk for severe fetal anemia due to anti-K. PMID- 11061863 TI - An amino acid substitution in the putative second extracellular loop of RBC band 3 accounts for the Froese blood group polymorphism. AB - BACKGROUND: The low incidence RBC antigen Fr(a) has been excluded from 17 of the 25 established blood group systems. Previous genetic analysis assigned the gene controlling Fr(a) expression to the same chromosomal region as the solute carrier family 4, anion exchanger member 1 gene (SLC4A1). Because SLC4A1 encodes RBC band 3 and controls the expression of Diego blood group system antigens, the possible relationship of Fr(a) to the Diego blood group system was investigated by molecular analysis of SLC4A1. STUDY DESIGN AND METHODS: Blood samples were obtained from the members of two unrelated Mennonite kindreds segregating for Fr(a). DNA was extracted, amplified by PCR using intronic primer sets flanking exons 11-20 of SLC4A1, and screened by single-strand conformation polymorphism (SSCP) analysis. Those exons displaying SSCPs were subjected to DNA sequence analysis. RESULTS: An exon 13 SSCP mobility shift was observed in the DNA from all Fr(a+) persons that was not seen in the DNA from Fr(a-) family members or control subjects. Linkage between the exon 13 SSCP and FR:(a) was established, with peak lods = 3.62 at theta = 0.00 for combined paternal and maternal meioses. DNA sequencing revealed a GAG --> AAG mutation that underlies a Glu480Lys substitution in RBC band 3. CONCLUSIONS: A point mutation in exon 13 of SLC4A1 accounting for a Glu480Lys substitution in band 3 controls Fr(a) expression. On the basis of these our results, the International Society of Blood Transfusion Working Party on Terminology for Red Cell Surface Antigens has assigned Fr(a) to the Diego blood group system, with the designation DI20. PMID- 11061864 TI - Human promyelocytic cell line: a convenient tool for studying the molecular basis of WBC filtration. AB - BACKGROUND: Blood filtration is a technique widely used to reduce the levels of WBCs in blood components. Several studies have been conducted to define the factors that are involved in WBC reduction, but the various mechanisms are not clearly delineated. This study explored the role of WBC adhesion molecules in WBC reduction during filtration. STUDY DESIGN AND METHODS: A minifilter has been developed that has properties similar to those of the standard filter (Sepacell, Asahi Medical) but that allows a smaller volume of blood to be used (15 mL). WBC reduction was achieved to a similar extent in the standard filter and the minifilter (4.15 log and 4.18 log, respectively). Samples of human promyelocytic cell line (HL60) were filtered before and after differentiation induced by vitamin D3 (D3-HL60). Flow cytometry was used to characterize the D3-HL60 filtrates and to count the WBCs after filtration. RESULTS: HL60 was retained in the filter to the same extent as all other WBCs. A higher level of integrin receptors (CD11b/CD18; CD11c/CD18) was expressed by D3-HL60 than by HL60. When the blood was incubated with anti-CD11b, anti-CD11c, or anti-CD18, fewer D3-HL60 cells were trapped by the filter, while only anti-CD11b alters HL60 retention in the filter. CONCLUSION: The receptors CD11b/CD18 and CD11c/CD18 appear to bind to the filter fibers and to be one of the mechanisms responsible for WBC retention. PMID- 11061865 TI - The effect of storage on the expression of platelet membrane phosphatidylserine and the subsequent impacton the coagulant function of stored platelets. AB - BACKGROUND: Platelet concentrates (PCs) derived from whole blood and stored under standard blood bank conditions undergo changes that are referred to as the platelet storage lesion. This study assesses the effect of PC preparation and storage on the distribution of phosphatidylserine (PS) in the platelet membrane and the effect that this distribution may have on the thrombogenic potential of stored PCs. STUDY DESIGN AND METHODS: Fresh platelets and PCs donated by healthy donors were obtained. PCs derived from platelet-rich plasma were studied on Day 1, Day 3, and Day 6 of storage under blood bank conditions. RESULTS: Platelet aggregation after exposure to the platelet agonists ADP and epinephrine singly declined progressively, but, when ADP and epinephrine in combination and collagen and thrombin in combination were used as agonists, the decline in platelet aggregation was less marked. PS expression as measured by Annexin V binding (mean and SD) was 2.02 +/- 0.93 percent in fresh platelet samples and increased to 5.39 +/- 4.2 percent on Day 1, 22. 1 +/- 7.1 percent on Day 3, and 39.5 +/- 12.1 percent on Day 6. Platelet prothrombinase activity (mean +/- SD) as measured by thrombin generation increased from 1.49 +/- 0.7 micro per mL in fresh platelet samples to 3.68 +/- 1.1 micro per mL in Day 1 platelets (p<0.001), 5.15 +/- 2.5 micro per mL in Day 3 platelets (p<0.001), and 4.65 +/- 2.48 micro per mL in Day 6 platelets (p<0. 001). CONCLUSION: These results show that PS expression increases after preparation of PCs from platelet-rich plasma and rises progressively during platelet storage under blood bank conditions. Furthermore, the greater PS expression is associated with increased platelet- dependent thrombin-generating capacity. PMID- 11061866 TI - Completely converting a blood service region to the use of safer plasma. AB - BACKGROUND: Three types of plasma are widely available for transfusion. Two plasma components, FFP donor retested (FFP-DR), and solvent/detergent-treated plasma (SDP), are now considered to be safer from infectious complications than FFP. STUDY DESIGN AND METHODS: A large regional blood center attempted to provide FFP-DR exclusively to all its 42 hospitals. Significant planning, increases in computer capabilities, and expansion of component storage areas were completed before initiation of this program. RESULTS: During the first 6 months of the FFP DR program, the blood center was not able to supply the entire region exclusively with FFP-DR. Consequently, SDP was utilized to supplement the program and to successfully and completely convert the region's 42 hospitals to the use of safer plasma. CONCLUSION: Two new plasma components were utilized to completely convert a blood service region to the use of safer plasma. PMID- 11061867 TI - Residues of NAT: questions and commentary. PMID- 11061868 TI - Summary of a workshop on the implementation of NAT to screen donors of blood and plasma for viruses. PMID- 11061869 TI - A novel cis-AB variant allele arising from a nucleotide substitution A796C in the B transferase gene. PMID- 11061870 TI - Spontaneous improvement of acute mixed lineage leukemia after blood transfusion. PMID- 11061871 TI - Platelet contamination and immunomagnetic cellular selection with the Isolex 300i cell-selection device. PMID- 11061872 TI - Prolonged antibody-negative HCV viremia in a US blood donor with apparent HCV transmission to a recipient. PMID- 11061873 TI - Pelvic prolapse. AB - PURPOSE: Pelvic prolapse is a common problem affecting women of all ages. We reviewed the pathophysiology, presentation, evaluation and treatment of pelvic prolapse. MATERIALS AND METHODS: We comprehensively reviewed the literature using MEDLINE, resources cited in those peer reviewed articles and abstracts from recent international meetings. RESULTS: Pelvic prolapse involves the herniation of various portions of the vaginal wall. Symptoms vary according to the area of the vagina affected. Proper evaluation is imperative for providing proper treatment. Various surgical approaches to repair have been developed and techniques continue to evolve. CONCLUSIONS: With the increasing involvement of urologists in the treatment of pelvic prolapse it is essential for us to become familiar with the anatomy, and the evaluation and management options available. We provide an overview of the care of patients with pelvic prolapse. PMID- 11061874 TI - A prospective analysis of time to normalization of serum testosterone after withdrawal of androgen deprivation therapy. AB - PURPOSE: Patients with prostate cancer are treated with neoadjuvant, adjuvant and intermittent androgen deprivation therapy. Prostate specific antigen (PSA) is altered during androgen deprivation therapy, and as a result the prognostic significance and accuracy of PSA values measured before serum testosterone has normalized are questionable because the patient is still effectively on androgen deprivation therapy. We determine the time it takes for serum testosterone to return to normal after withdrawal of androgen deprivation therapy. MATERIALS AND METHODS: Serial serum testosterone was prospectively measured at 3-month intervals in 68 men after withdrawal of androgen deprivation therapy. The number of months to return to normal serum testosterone 270 ng./dl. or greater, was calculated for each patient. Patients were stratified according to duration of androgen deprivation, age and type of luteinizing hormone releasing hormone agonist used. RESULTS: Median patient age was 71 years (range 46 to 88). Median time to normalization of testosterone was 7 months (range 1 to 58). At 3, 6 and 12 months 28%, 48% and 74% of men had normal serum testosterone, respectively. Serum testosterone took significantly longer to return to normal in patients on androgen deprivation therapy for 24 months or greater compared to those on therapy for less than 24 months (log-rank p = 0.0034). There was no statistical significance based on age or type of luteinizing hormone releasing hormone agonist used. CONCLUSIONS: Androgen deprivation has an effect on serum testosterone that extends beyond the cessation of treatment. Serum testosterone should be measured in all men until normalization. These results should be applied to the interpretation of PSA levels after withdrawal of androgen deprivation therapy. In addition, these data have implications regarding dose scheduling and definition of biochemical (PSA) failure after primary therapy. PMID- 11061875 TI - An anterior extraperitoneal incision for donor nephrectomy that spares the rectus abdominis muscle and anterior abdominal wall nerves. AB - PURPOSE: To perform open donor nephrectomy with the least amount of body alteration and complication, a short extraperitoneal incision was developed that preserves the ipsilateral rectus abdominis muscle and abdominal branches of the intercostal nerves. MATERIALS AND METHODS: Donor nephrectomy using the new incision was performed in 11 consecutive patients. RESULTS: Morphine was required for pain relief for 1 to 3 days (mean 1.8), patients tolerated diet at 1 to 4 days (mean 2.2) and postoperative stay was 2 to 5 days (mean 3.5). Complications were minimal, consisting of postoperative fever 1 day in duration. A viable kidney was provided for transplantation in each case. CONCLUSIONS: The extraperitoneal, rectus abdominis muscle and nerve sparing incision resulted in a short course of pain medication, early diet toleration and a short hospital stay. PMID- 11061876 TI - Surveillance of upper urinary tract transitional cell carcinoma: the role of ureteroscopy, retrograde pyelography, cytology and urinalysis. AB - PURPOSE: A select group of patients with upper tract transitional cell carcinoma are treated with ureteroscopic resection. We determine the validity and accuracy of urinalysis, bladder cytology, upper tract biopsy/cytology and retrograde pyelography for the detection of recurrent upper tract transitional cell carcinoma compared to endoscopic findings. MATERIALS AND METHODS: Patients with ureteroscopically treated upper tract transitional cell carcinoma were followed with surveillance every 3 to 6 months. Surveillance included urinalysis with dipstick and microscopic examination, bladder cytology, retrograde pyelography read by a urologist and radiologist, and ureteropyeloscopy with cytology and biopsy of suspicious areas. Not all results were available for all surveillance procedures. Measures of sensitivity and specificity for the aforementioned surveillance procedures were determined relative to endoscopic findings that were defined as the standard. Confidence intervals were also estimated. Initially, a generalized estimation equation approach was used to take into account the clustering of repeated testing within patients. The accuracy of each procedure was also calculated. RESULTS: There were 23 patients with previously resected low grade upper tract transitional cell carcinoma who underwent a total of 88 surveillances in 30 months. A total of 56 of 88 (64%) recurrences were detected ureteroscopically, including 11 (12%) associated bladder recurrences. In patients who did not have bladder recurrences urinalysis had a sensitivity of 37.5% but specificity was 85%, while bladder cytology had a sensitivity of 50% and specificity was 100%, and retrograde pyelography read in the endoscopy room revealed a sensitivity of 71.7% and specificity of 84.7%. Ureteroscopic biopsy/cytology had a sensitivity and specificity of 93.4% and 65.2%, respectively. CONCLUSIONS: Our findings indicate that compared to ureteroscopy, urinalysis, bladder cytology, retrograde pyelography and ureteroscopic cytology/biopsy are less valid and accurate in detecting upper tract transitional cell carcinoma recurrences. Based on our data we recommend ureteroscopic evaluation as an essential procedure for the surveillance of patients treated endoscopically for upper tract transitional cell carcinoma. PMID- 11061877 TI - Low success rate of repeat shock wave lithotripsy for ureteral stones after failed initial treatment. AB - PURPOSE: We determined the number of shock wave lithotripsy treatments that should be given for a single ureteral stone before alternate modalities are used. MATERIALS AND METHODS: We compared the stone-free rate of initial shock wave lithotripsy for ureteral calculi with that of subsequent treatments. We evaluated 1,593 ureteral stones treated with the Dornier MFL 5000 lithotriptor* from January 1, 1994 to September 1, 1999 using various parameters associated with treatment outcome. RESULTS: The stone-free rate after initial treatment was 68% (1,086 of 1,593 stones), which decreased to 46% (126 of 273) after re-treatment 1. We observed a further decrease in the stone-free rate after re-treatment 2 to 31% (19 of 61 stones, p = 0.001). The cumulative stone-free rate increased to 76% (1,212 of 1,593 stones) after 2 treatments and to 77% (1,231 of 1593) after 3. The stone-free rate for stones 10 mm. or less was significantly better than that of stones 11 to 20 mm. initially (64% versus 43%) and after re-treatment (49% versus 37%). A ureteral stent decreased the stone-free rate of initial treatment and re-treatment 1 by 12% and 14%, respectively (p = 0.001). After initial treatment the stone-free rate of the upper and mid ureter was significantly higher than that of the lower ureter. Patient weight had no significant impact on success in either group. CONCLUSIONS: The stone-free rate of re-treating ureteral calculi with shock wave lithotripsy decreases significantly after the initial treatment. These findings imply that ureteroscopic management of ureteral stones may be better than shock wave lithotripsy after initial shock wave lithotripsy fails. PMID- 11061878 TI - Comparison of cystoscopic and histological findings in patients with suspected interstitial cystitis. AB - PURPOSE: Although the exact etiology of interstitial cystitis remains elusive, bladder inflammation appears to be common in many patients. The National Institutes of Health (NIH) have established diagnostic criteria for interstitial cystitis based on the presence of irritative voiding symptoms in the absence of other identifiable pathology. Cystoscopic examination with hydrodistention performed in patients under anesthesia is part of the NIH diagnostic criteria. We determine if the severity of cystoscopic findings correlated with histological evidence of inflammation in patients with suspected interstitial cystitis. MATERIALS AND METHODS: A total of 69 patients who met NIH symptom criteria for interstitial cystitis and underwent cystoscopy, hydrodistention and bladder biopsy under anesthesia were reviewed. There were 2 investigators blinded to the histological data who independently reviewed operative reports. A urological pathologist blinded to the clinical data reviewed biopsies for inflammation severity. Cystoscopic and histological findings were then converted to a numeric scale. Numeric data were analyzed using the Pearson correlation coefficient. RESULTS: Cystoscopic examination revealed no evidence of interstitial cystitis in 6 patients (9%), mild changes in 27 (39%), moderate changes in 23 (33%) and severe changes in 13 (19%). Histological examination revealed no inflammation in 21 patients (30%), mild inflammation in 28 (41%), moderate inflammation in 11 (16%) and severe inflammation in 9 (13%). Histological scores correlated poorly with total and scaled cystoscopic severity scores (r = 0.295 and 0.349, respectively). CONCLUSIONS: Severity of cystoscopic findings observed during hydrodistention with anesthesia does not appear to correlate with the degree of inflammation identified histologically in patients with suspected interstitial cystitis. PMID- 11061879 TI - Intravesical bacillus Calmette-Guerin and dimethyl sulfoxide for treatment of classic and nonulcer interstitial cystitis: a prospective, randomized double blind study. AB - PURPOSE: We conducted a prospective, double-blind study with a crossover design of intravesical bacillus Calmette-Guerin (BCG) and dimethyl sulfoxide to determine whether patients with classic and nonulcer interstitial cystitis, respectively, might benefit from either regimen. MATERIALS AND METHODS: A total of 21 patients, including 11 with classic and 10 with nonulcer interstitial cystitis, randomly underwent treatments with intravesical BCG or dimethyl sulfoxide and, if not improved, were treated with the other substance after a washout period. All 21 patients were evaluated with symptom questionnaires, including a visual analog pain scale and voiding diaries. RESULTS: Regardless of regimen, there was no improvement in maximal functional capacity. There was a reduction in urinary frequency following dimethyl sulfoxide treatment but only in the classic subtype (p <0.05), whereas no reduction was seen following BCG in either subtype. A substantial pain decrease was noted in classic (p <0.05) as well as nonulcer (p <0.05) interstitial cystitis following dimethyl sulfoxide. CONCLUSIONS: Intravesical BCG has been presented as a promising new option for treatment of interstitial cystitis. We failed to demonstrate benefit from this treatment. Dimethyl sulfoxide had no positive effect on maximal functional capacity but resulted in a significant reduction in pain and urinary frequency, although only in patients with classic interstitial cystitis. PMID- 11061880 TI - Improvement of interstitial cystitis symptoms and problems that developed during treatment with oral IPD-1151T. AB - PURPOSE: We examined the efficacy of Suplatast Tosilatedouble dagger (IPD-1151T), a new immunoregulator that suppresses helper T cell mediated allergic responses, including IgE production and eosinophilic inflammation for treating patients with interstitial cystitis. MATERIALS AND METHODS: A total of 14 women (average age 43.7 years) with interstitial cystitis, which was nonulcerative in 13 and ulcerative in 1, were treated with 300 mg. IPD-1151T orally daily for 12 months. All patients received laboratory assessments, including hematology (eosinophils and CD20 positive cells) and serum chemistry (IgE, and interleukin-4 (IL-4) and 5, and immunohistochemical analyses of urine leukocytes (CD45RO positive cells as a T cell marker) before treatment. These parameters were also measured 4 and 12 months after continuous treatment. The voiding chart, and interstitial cystitis symptom and problem indexes were evaluated before and after IPD-1151T treatment. RESULTS: IPD-1151T treatment for 1 year resulted in a significantly increased bladder capacity and decreased symptoms, such as urinary urgency, frequency and lower abdominal pain, in patients with nonulcerative interstitial cystitis. These effects also correlated with a reduction in blood eosinophils, CD20 positive cells and IgE, and urine CD45RO positive memory T cells. No major side effects were observed. CONCLUSIONS: Our study suggests that immunological responses are involved in the development of interstitial cystitis symptoms. IPD-1151T could be a new oral agent for treatment of voiding symptoms and bladder pain in patients with interstitial cystitis. PMID- 11061881 TI - Interstitial cystitis--the great enigma. PMID- 11061882 TI - Immunostaining of cytokeratin 20 in cells from voided urine for detection of bladder cancer. AB - PURPOSE: Cytokeratin 20 is a cytoskeletal protein expressed in colon and bladder cancer cells but only rarely in normal urothelium. Previous studies have shown that identification of RNA coded for cytokeratin 20 in urine samples using polymerase chain reaction is highly sensitive for detection of bladder tumors. We examined the efficacy of immunocytology on cells from voided urine samples using monoclonal antibodies against cytokeratin 20 for noninvasive detection of bladder tumors. MATERIALS AND METHODS: A total of 174 patients comprised the study, including 80 who were evaluated because of hematuria or irritative voiding symptoms and 94 who were examined during followup visits after resection of bladder tumors. Voided urine samples were obtained for immunocytology and cytopathology. Each patient underwent cystoscopy, and biopsies were done when a bladder tumor was detected or carcinoma in situ was suspected. Indirect immunoperoxidase staining was done on cytocentrifuge slides using a monoclonal antibody against cytokeratin 20. RESULTS: Bladder tumors were found in 87 patients. Immunocytology resulted in 81.6% sensitivity, 77% specificity and 80% accuracy. False-negative results occurred in 12 patients with superficial low grade tumors and in 4 with high grade invasive tumors. Cytopathology resulted in 51.7% sensitivity and 94.2% specificity. CONCLUSIONS: Immunocytology of the cytokeratin 20 antigen is significantly more sensitive than cytopathology for bladder tumor detection, especially low grade and low stage cancer. The results are not affected by inflammation or previous treatments with bacillus Calmette Guerin. Immunocytology may miss high grade tumors and, therefore, cannot be used alone as a substitute for cystoscopy. PMID- 11061883 TI - Urinary level of nuclear matrix protein 22 in the diagnosis of bladder cancer: experience with 130 patients with biopsy confirmed tumor. AB - PURPOSE: We prospectively evaluated the value of nuclear matrix protein 22 (NMP22dagger) and cytology in the diagnosis of bladder cancer. MATERIALS AND METHODS: We analyzed NMP22 in voided urine from 235 patients before cystoscopy. Of the patients 130 had transitional cell carcinoma of the bladder and subsequently underwent surgery. In a subset of 200 patients bladder washout samples for cytology were collected during cystoscopy. The cutoff for NMP22 was 10.0 units per ml. For cytology only high grade atypia was considered positive. RESULTS: Histology showed 77 superficial (pTa, pTis) and 53 invasive (pT1 or greater) tumors. Sensitivity of NMP22 was 51% and specificity was 83%. NMP22 sensitivity was 36% for superficial tumors and 73% for invasive transitional cell carcinoma. Overall sensitivity of cytology was 52% and specificity was 89%. Cytology sensitivity was 38% for superficial tumors and 83% for invasive transitional cell carcinoma. NMP22 sensitivity for grades 1, 2 and 3 tumors was 30%, 56% and 68%, respectively. Cytology sensitivity for grades 1, 2 and 3 tumors was 30%, 50% and 91%, respectively. Combined NMP22 and cytology had a sensitivity of 70%. CONCLUSIONS: NMP22 has sensitivity and specificity similar to those of cytology from bladder washout samples. Particularly in low stage and low grade tumors both tests show the same disappointing sensitivity. Because of a false negative rate of 49%, NMP22 cannot replace cystoscopy in clinical practice, as the danger of missing NMP22 negative tumors is too high to rely on its results in an individual patient. PMID- 11061884 TI - Factors predicting recovery of erections after radical prostatectomy. AB - PURPOSE: Because preservation of functioning penile erections is a major concern for many patients considering treatment for localized prostate cancer, we analyzed various factors determined before and after radical retropubic prostatectomy to identify those significantly associated with recovery of erectile function. MATERIALS AND METHODS: Our prospective database of patients undergoing pelvic lymphadenectomy and radical retropubic prostatectomy was used to determine factors predictive of erection recovery after radical prostatectomy. The study included 314 consecutive men with prostate cancer treated with radical retropubic prostatectomy between November 1993 and December 1996. Preoperative potency satisfactory for intercourse and degree of neurovascular bundle preservation during the operation were documented. RESULTS: Patient age, preoperative potency status and extent of neurovascular bundle preservation but not pathological stage were predictive of potency recovery after radical prostatectomy. At 3 years after the operation 76% of men younger than age 60 years with full erections preoperatively who had bilateral neurovascular bundle preservation would be expected to regain erections sufficient for intercourse. Compared to the younger men, those 60 to 65 years old were only 56% (95% confidence interval [CI] 37 to 84) and those older than 65 years were 47% (95% CI 30 to 73) as likely to recover potency. Patients with recently diminished erections were only 63% (95% CI 38 to 100) as likely to recover potency as men with full erections preoperatively, and those with partial erections were only 47% (95% CI 23 to 96) as likely to recover potency. Resection of 1 neurovascular bundle reduced the chance of recovery to 25% (95% CI 10 to 61) compared to preserving both nerves. CONCLUSIONS: Knowledge of preoperative erectile function and patient age before the operation and the degree of neurovascular bundle preservation afterward may aid in patient counseling regarding potency recovery after radical prostatectomy. PMID- 11061885 TI - Effect of sildenafil citrate on post-radical prostatectomy erectile dysfunction. AB - PURPOSE: We assess the effect of sildenafil in a subgroup of patients after prostatectomy with erectile dysfunction and determine whether nerve preservation improves sildenafil response in this subgroup. MATERIALS AND METHODS: Between April 1998 and January 1999, 53 patients who had undergone radical retropubic prostatectomy and were prescribed oral sildenafil were surveyed using a confidential mail questionnaire. Of the patients 21 underwent bilateral and 15 unilateral neurovascular bundle sparing procedures, while in 17 a nonnerve sparing procedure was performed. All patients received 25 to 100 mg. sildenafil in a flexible dose escalation manner. Response, satisfaction and side effects were assessed using a modified, self-administered International Index of Erectile Function questionnaire. Response was defined as erection sufficient for intercourse. Preoperative and postoperative/pretreatment erectile functions were assessed for baseline comparison in each patient, and partner overall satisfaction with sildenafil was measured. Statistical data analysis was performed using analysis of variance and Newman-Keuls multiple comparison tests. RESULTS: Of the 21 patients who underwent a bilateral nerve sparing procedure 15 had a positive response. Of the 15 patients who had undergone a unilateral nerve sparing procedure 12 had a positive response, and only 1 of the 17 patients who had undergone a nonnerve sparing procedure responded to sildenafil. The most commonly reported adverse events of all causes were headaches (21%), flushing (8.3%), visual disturbance (6.3%) and nasal congestion (6.3%). CONCLUSIONS: Sildenafil is an equally effective treatment for erectile dysfunction after bilateral and unilateral nerve sparing procedures, and patient response to sildenafil is confirmed by the partners. However, patients who undergo nonnerve sparing procedures do not respond satisfactorily to sildenafil. PMID- 11061886 TI - Vasectomy reversal for the post-vasectomy pain syndrome: a clinical and histological evaluation. AB - PURPOSE: The cause of the post-vasectomy pain syndrome is unclear. Some postulated etiologies include epididymal congestion, tender sperm granuloma and/or nerve entrapment at the vasectomy site. To our knowledge nerve proliferation has not been evaluated previously as a cause of pain. Vasectomy reversal is reportedly successful for relieving pain in some patients. We report our experience and correlate histological findings in resected vasal segments with outcome to explain the mechanism of pain in these patients. MATERIALS AND METHODS: We retrospectively reviewed the records of 13 men who underwent vasectomy reversal for the post-vasectomy pain syndrome. We compared blinded histological evaluations of the vasal ends excised at vasectomy reversal in these patients with those of pain-free controls who underwent vasectomy reversal to reestablish fertility. Controls were matched to patients for the interval since vasectomy. Histological features were graded according to the degree of severity of vasitis nodosum, chronic inflammation and nerve proliferation. RESULTS: Mean time to pain onset after vasectomy was 2 years. Presenting symptoms included testicular pain in 9 cases, epididymal pain in 2, pain at ejaculation in 4 and pain during intercourse in 8. Physical examination demonstrated tender epididymides in 6 men, full epididymides in 6, a tender vasectomy site in 4 and a palpable nodule in 4. No patient had testicular tenderness on palpation. Unilateral and bilateral vasovasostomy was performed in 3 and 10 of the 13 patients, respectively. Postoperatively 9 of the 13 men (69%) became completely pain-free. Mean followup was 1.5 years. We observed no differences in vasectomy site histological features in patients with the post-vasectomy pain syndrome and matched controls, and no difference in histological findings in patients with the post-vasectomy pain syndrome who did and did not become pain-free postoperatively. CONCLUSIONS: No histological features aid in identifying a cause of pain or provide prognostic value for subsequent pain relief. Vasectomy reversal appeared to be beneficial for relieving pain in the majority of select patients with the post-vasectomy pain syndrome. PMID- 11061887 TI - Clinical stage I pure yolk sac tumor of the testis in adults has different clinical behavior than juvenile yolk sac tumor. AB - PURPOSE: We evaluated the clinical behavior of clinical stage I pure yolk sac tumor of the testis in adults to determine whether the behavior of this entity is different than that of clinical stage 1 nonseminoma. MATERIALS AND METHODS: We searched the testis cancer database at our institution for adults with clinical stage I pure yolk sac tumor of the testis who underwent retroperitoneal lymph node dissection. We identified 12 such patients and reviewed the database and hospital charts to determine clinical behavior. RESULTS: Disease was pathological stage I in 8 of the 12 patients (66%), including 1 with recurrence after retroperitoneal lymph node dissection. Disease was pathological stage II in 14 patients (33%), including 1 who remains disease-free after electing adjuvant bleomycin, etoposide and cisplatin. Of the 3 patients who elected observation after retroperitoneal lymph node dissection only 1 has had recurrence, while 2 (66%) were cured by retroperitoneal lymph node dissection only. CONCLUSIONS: Contrary to juvenile yolk sac tumor, which has a strong tendency toward hematogenous metastasis, the behavior of clinical stage I adult pure yolk sac tumor is similar to that of all other stage I nonseminomas in adulthood. PMID- 11061888 TI - Molecular epidemiological evidence for ascending urethral infection in acute bacterial prostatitis. AB - PURPOSE: To test the ascending urethral infection in the pathogenesis of acute bacterial prostatitis, we assessed the clonality of Escherichia coli strains isolated from urine and rectal swab of patients with acute bacterial prostatitis using molecular typing methods. MATERIALS AND METHODS: A total of 50 E. coli strains each isolated from urine and rectal swabs of 9 men with acute bacterial prostatitis at diagnosis were examined for 6 urovirulence determinant profiles and pulsed field gel electrophoresis patterns. In 1 case E. coli isolates from the rectal swab of the patient's wife were also examined at diagnosis and after 5 weeks. RESULTS: The urovirulence profile and pulsed field gel electrophoresis demonstrated that causative E. coli was monoclonal in each case, and present in the rectal swab as a predominant (96% to 100%) fecal clone in 2 and a minority clone (2% to 8%) in 4. Furthermore, causative E. coli dominated in the rectal swab of the 1 patient's wife. CONCLUSIONS: Our results are consistent with the ascending route of infection in acute bacterial prostatitis. However, causative E. coli might possibly originate from either intestinal reservoir of the host or household member. Owing to limitations of the cross-sectional design of this study, longitudinal studies are necessary to establish the ascending route of infection in this disease. PMID- 11061889 TI - Scoring the short form ICSmaleSF questionnaire. International Continence Society. AB - PURPOSE: The International Continence Society (ICS) ICSmale questionnaire was devised to provide a thorough evaluation of the occurrence and bothersomeness of lower urinary tract symptoms and their impact on the lives of men with benign prostatic disease. This report completes the developmental work on the questionnaire, producing the concise short form instrument, ICSmaleSF, with a valid, reliable and scientifically justified scoring system. MATERIALS AND METHODS: Two data sets were used. The short form version of the questionnaire was devised and initially evaluated using data on men with uncomplicated lower urinary tract symptoms who were involved in the CLasP randomized controlled trial comparing laser therapy with transurethral prostatic resection and conservative management or monitoring without active intervention. External validation of the scoring system was undertaken using data from phase II of the ICS benign prostatic hyperplasia (BPH) study, an observational study of outcome in men with lower urinary tract symptoms related to benign prostatic enlargement. All patients completed the developmental version of the ICSmale questionnaire. Parallel analysis on the CLasP data set identified items that were responsive to change or highly problematic, allowing other redundant and overlapping items to be eliminated. Factor analysis and Cronbach's alpha coefficients were used to examine the clustering of items. Regression models were used to investigate the validity of followup scores within and across treatment groups in the CLasP and ICS/BPH studies. RESULTS: The questionnaire, which originally comprised 22 items, was shortened to 11 items in the 2 distinct factors of voiding (ICSmaleVS) and incontinence (ICSmaleIS) symptoms. Cronbach's alpha coefficients were high at 0.76 for ICSmaleVS and 0.78 for ICSmaleIS. A simple additive score was calculated by adding the 5 items in ICSmaleVS and 6 in ICSmaleIS. ICSmaleVS and ICSmaleIS detected expected improvement in the laser therapy and transurethral prostatic resection groups, and stability in the conservative management group within CLasP. Similarly each subscore but particularly ICSmaleVS was sensitive to differences in the outcome of the range of treatments in the ICS/BPH study. While frequency and nocturia were highly problematic and sensitive to change individually, they did not load into the other main factors or correlate with each other. It is suggested that these symptoms should be evaluated separately with the additional inclusion of a single item measure of the impact of lower urinary tract symptoms on daily life. CONCLUSIONS: The ICSmaleSF represents a comprehensive, concise, valid and reliable instrument for evaluating men with lower urinary tract symptoms. Unlike other questionnaires in the field it contains subscores for the domains of voiding and incontinent symptoms as well as the separate consideration of frequency, nocturia and impact on daily life. We hope that it will become the tool of choice for the comprehensive evaluation of treatment of men with lower urinary tract symptoms associated with benign prostatic disease. PMID- 11061890 TI - Use of the complex between prostate specific antigen and alpha 1-protease inhibitor for screening prostate cancer. AB - PURPOSE: We assess whether the complex between prostate specific antigen (PSA) and alpha1-protease inhibitor in serum can be used to reduce further the number of false-positive PSA screen results independent of total and free PSA. MATERIALS AND METHODS: Sera from 304 consecutive screen positive subjects, including 78 with and 226 without prostate cancer, and serum PSA of 4 to 10 microg./l. or higher in the Finnish, randomized, population based prostate cancer screening trial were analyzed for PSA-alpha-protease inhibitor, and total and free PSA. Main outcome measures were specificity, sensitivity and area under receiver operating characteristics curve for proportions of free PSA and PSA-alpha 1 protease inhibitor, and for a combination of these among screen positive cases. RESULTS: The proportion of serum PSA-alpha 1-protease inhibitor of total PSA was lower in cancer cases than in controls (0.9% versus 1.6%, p <0.001). Logistic regression analysis of total PSA, free PSA and PSA-alpha 1-protease inhibitor showed that PSA-alpha 1-protease inhibitor in serum was an independent variable for discrimination between subjects with and without prostate cancer (p = 0.006) in the PSA range of 4 to 10 microg./l. The proportion of PSA-alpha 1-protease inhibitor alone improved specificity less than the proportion of free PSA but when these were combined by logistic regression they performed better than the proportion of free PSA alone at sensitivities of 85% to 95% (p <0.001). CONCLUSIONS: Serum PSA-alpha 1-protease inhibitor improves the specificity of total and free PSA in a screening population with total PSA 4 to 10 microg./l. PMID- 11061891 TI - Prostate cancer detection in Black and White men with abnormal digital rectal examination and prostate specific antigen less then 4 ng./ml. AB - PURPOSE: Prostate cancer is more common in black than in white American men. Experience in a longitudinal prostate cancer screening program implies that cancer detection is greater in black than in white men with an abnormal digital rectal examination and prostate specific antigen (PSA) less than 4 ng./ml. We investigated potential racial differences in cancer detection in men treated in clinical practice who had an abnormal digital rectal examination and PSA less than 4 ng./ml. MATERIALS AND METHODS: Between January 1992 and December 1999 prostate biopsy was done at a Veterans Affairs Medical Center in 179 black and 357 white men with an abnormal digital rectal examination, PSA less than 4 ng./ml. and no history of prostate surgery. Significant racial differences in demographic and clinical parameters were limited to PSA, which was higher in black men (p = 0.01). RESULTS: Cancer was detected in 38 black (21%) and 65 white (18%) men (p = 0.42). There were no significant racial differences in the PSA adjusted cancer detection rate or in the Gleason score of detected disease. In men with PSA less than 1.0, 1.0 to 1.9, 2.0 to 2.9 and 3.0 to 3.9 ng./ml. the detection rate was 4%, 15%, 27% and 29%, respectively. CONCLUSIONS: In clinical practice prostate cancer detection appears to be equivalent in black and white men when an abnormal digital rectal examination is the only indication of malignancy. PMID- 11061892 TI - Prostate cancer in men age 50 years or younger: a review of the Department of Defense Center for Prostate Disease Research multicenter prostate cancer database. AB - PURPOSE: Prostate cancer in men age 50 years or younger traditionally has accounted for approximately 1% of those diagnosed with prostate cancer. Prior studies of prostate cancer in men of this age led many clinicians to believe that they have a less favorable outcome than older men. Most of these studies were conducted before the advent of prostate specific antigen (PSA) screening programs. We evaluated a surgically treated cohort of men age 50 years or younger to determine whether disease recurred more frequently among them than in those 51 to 69 years old in the PSA era. MATERIALS AND METHODS: We reviewed the medical records of 477 men who underwent radical prostatectomy between 1988 and 1997. Age, ethnicity, preoperative PSA, clinical and pathological stage, margin and seminal vesicle involvement, and recurrence were compared between 79 men age 50 years or younger (study group) and 398, 51 to 69 years old (comparison group). Disease-free survival rates were compared using Kaplan-Meier and Cox regression techniques. RESULTS: There were 6 (7.6%) recurrences in the study group (79) and 107 (26.9%) in the comparison group (398). The disease-free survival curves were significantly different (log-rank p = 0.010). Age remained a significant prognostic factor (Wald p = 0.033) in multivariate Cox regression analyses that controlled for race, clinical and pathological stage, and pretreatment PSA. Similar results were found when the comparison group was limited to 116 patients 51 to 59 years old (log-rank p = 0.034, Wald p = 0.069). CONCLUSIONS: These data suggest that patients in the PSA era who underwent radical prostatectomy and were age 50 years or younger have a more favorable disease-free outcome compared to older men. PMID- 11061893 TI - Is preoperative donation of autologous blood rational for radical retropubic prostatectomy? AB - PURPOSE: We investigated differences in the rate of homologous blood transfusion and the degree of anemia to determine whether it is rational to have patients donate autologous blood before radical prostatectomy. MATERIALS AND METHODS: We retrospectively reviewed the charts of 221 consecutive men who underwent radical retropubic prostatectomy performed by 1 surgeon in a 14-month period. About half of the patients donated autologous blood preoperatively. We evaluated perioperative hemoglobin, and the rate of autologous and homologous transfusion. RESULTS: The groups did not significantly differ in terms of demographic data, co morbid conditions, clinical variables or hospitalization. Preoperatively mean hemoglobin plus or minus standard deviation was 13.4 +/- 1 and 14.7 +/- 1 gm./dl. in patients who did and did not donate blood, while homologous transfusion was required in 1 (1%) and 4 (3.5%), respectively (p = 0. 18). At hospital discharge anemia was more prevalent in nondonors. Of the men who did versus did not donate blood hemoglobin was less than 10 and less than 9 gm./dl. in 8.4% versus 34% (p <0.0001), and 12.5% versus 0% (p <0.0004), respectively. CONCLUSIONS: Our retrospective review of a cohort of patients who underwent radical retropubic prostatectomy showed no difference in homologous blood transfusion based on preoperative autologous donation status. Autologous donors had lower hemoglobin preoperatively, a higher rate of transfused units and higher hemoglobin at hospital discharge. Preoperative donation of autologous blood may not decrease the need for homologous transfusion in healthy patients undergoing radical retropubic prostatectomy. PMID- 11061894 TI - Urinary function and bother after radical prostatectomy or radiation for prostate cancer: a longitudinal, multivariate quality of life analysis from the Cancer of the Prostate Strategic Urologic Research Endeavor. AB - PURPOSE: We measure the effect of time on urinary function and bother during the first 2 years following treatment for early stage prostate cancer. MATERIALS AND METHODS: We studied urinary function and bother in 564 men recently diagnosed with early stage prostate cancer and treated with radiotherapy or radical prostatectomy with or without nerve sparing. Outcomes were assessed with the UCLA Prostate Cancer Index, which is a validated, health related quality of life instrument that includes these 2 domains. To minimize the influence of other factors we adjusted for age, co-morbidity, general health, pad use, anticholinergics or procedures for urethral stricture. All subjects were drawn from the Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE), which is a national longitudinal database. RESULTS: Urinary function improved with time during the first year after surgery but remained fairly constant during year 2. Urinary function remained stable throughout the 2 years after radiation. Urinary bother was worse after radiation throughout the 2 years, although it improved markedly by the end of year 1. Age, ethnicity and co-morbidity did not impact urinary function or bother but being married did have an advantage. CONCLUSIONS: Patients undergoing surgery or radiation showed different longitudinal profiles of urinary function and bother during the first 2 years after treatment. PMID- 11061895 TI - Major surgery to manage definitively severe complications of salvage cryotherapy for prostate cancer. AB - PURPOSE: Severe complications of salvage cryotherapy may be debilitating and chronic but these complications may be managed by definitive extirpative surgical procedures. We evaluated the effectiveness of the major surgical procedures performed to manage these complications, and assessed patient survival and complications after extirpative surgery. MATERIALS AND METHODS: Between 1992 and 1995 salvage cryotherapy was performed in 150 men with biopsy proved, locally recurrent prostate cancer after radiotherapy and/or systemic therapy. We retrospectively reviewed patient charts to assess the complications managed by extirpative surgery. RESULTS: Extirpative surgery was performed in 6 of the 150 patients for serious complications, including uncontrollable hematuria, osteitis pubis, rectourethral fistula, refractory perineal pain, bladder outlet obstruction and complete urinary incontinence. Cystoprostatectomy was done in 4 patients, of whom 3 also underwent en bloc pubic symphysectomy. In the remaining 2 men salvage prostatectomy was performed with bladder neck closure and continent catheterizable stomal creation. Surgery successfully managed severe cryotherapy complications in all 6 cases. The complications of extirpative surgery included superficial wound infection in 1 patient and 3 incisional hernias in another. Prostate specific antigen was undetectable in 4 of the 6 men at 36, 38, 39 and 42 months, and detectable in 2 at 31 and 41 months, respectively. CONCLUSIONS: Extirpative surgery may successfully alleviate severe salvage cryotherapy complications without major additive morbidity. Long survival duration justifies extirpative surgery in select patients with severe complications of salvage cryotherapy. PMID- 11061896 TI - The extent of biopsy involvement as an independent predictor of extraprostatic extension and surgical margin status in low risk prostate cancer: implications for treatment selection. AB - PURPOSE: We identify predictors of extraprostatic extension and positive surgical margins in patients with low risk prostate cancer (prostate specific antigen [PSA] 10 ng./ml. or less, biopsy Gleason score 7 or less and clinical stage T1c 2b). MATERIALS AND METHODS: From August 1997 to January 1999, 143 previously untreated patients underwent radical retropubic prostatectomy for clinically localized prostate cancer. A total of 62 patients were low risk, with PSA 10 ng./ml. or less, biopsy Gleason score 7 or less and clinical stage T1c-2b, and had sextant biopsy with separate pathological evaluation of each sextant cores. PSA, clinical stage, biopsy Gleason score, average percentage of cancer in the entire biopsy specimen, maximum percentage of cancer on the most involved core, number of cores involved and bilaterality were evaluated for association with extraprostatic extension, seminal vesicle involvement and positive surgical margins. RESULTS: Of the 62 patients 13 (21%) had extraprostatic extension, 6 (10%) seminal vesicle involvement and 20 (32%) positive surgical margins. Average percentage greater than 10% and maximum percentage greater than 25% were associated with extraprostatic extension (p = 0.01 and 0.004, respectively). Average percentage greater than 10%, maximum percentage greater than 25%, more than 2 cores involved and bilaterality were associated with positive surgical margins (p = 0.007, 0.01, 0.002 and 0.03, respectively). On multivariate analysis maximum percentage remained the only independent predictor of extraprostatic extension (p = 0.03), and the number of cores involved remained an independent predictor of positive surgical margins (p = 0.01). Biopsy Gleason score, PSA and clinical stage did not correlate with extraprostatic extension or positive surgical margins in this patient population. CONCLUSIONS: In low risk prostate cancer the extent of biopsy involvement significantly correlates with the risk of extraprostatic extension and positive surgical margins. Biopsy information should be considered when selecting and modifying treatment modalities. PMID- 11061897 TI - Preoperative prostate needle biopsy p27 correlates with subsequent radical prostatectomy p27, Gleason grade and pathological stage. AB - PURPOSE: Loss of p27 protein expression in radical prostatectomy specimens has been shown to be an adverse prognostic factor in patients with clinically localized prostate cancer. To our knowledge no studies have examined p27 expression in prostate needle biopsies. To test the potential predictive power of p27 in prostate biopsies we compared p27 expression in preoperative biopsies and matched prostatectomy specimens of patients with clinically localized prostate cancer. MATERIALS AND METHODS: Matched biopsies and radical prostatectomy specimens from 44 patients were examined. Mean followup was 22.7 months (range 1 to 46). Tumors expressing less than 30% positive nuclei were classified as low expressors and tumors expressing greater than 30% positive nuclei were classified as high expressors of p27 protein. RESULTS: Expression of p27 in prostate biopsies correlated significantly with subsequent p27 expression in radical prostatectomy specimens (p = 0.002). Sensitivity and specificity of biopsy p27 for predicting subsequent prostatectomy p27 were 87.5% and 88.9%, respectively (p <0.001). Univariate analysis showed that low expression of p27 in the biopsy correlated significantly with biopsy and prostatectomy Gleason score (p = 0.000 and 0.001, respectively), and final pathological stage (p = 0.028). Despite the small sample size and short followup, 36.4% of patients with low p27 expression had a biochemical recurrence compared to only 12.1% with high expression (hazards ratio 3.56). In addition, Kaplan-Meier analysis suggested that low p27 expression in prostate biopsies may be associated with a shorter time to recurrence, although this did not reach statistical significance (p = 0.081). CONCLUSIONS: Expression of p27 in prostate biopsies can be used to predict the degree of expression in radical prostatectomy specimens. As loss of p27 protein expression in prostatectomy specimens has been shown to correlate with biochemical recurrence and shortened prostate specific survival, these results suggest that biopsy p27 may help identify high risk patients preoperatively. PMID- 11061898 TI - Androgen receptor gene amplification at primary progression predicts response to combined androgen blockade as second line therapy for advanced prostate cancer. AB - PURPOSE: Amplification of the androgen receptor gene has been found in a third of hormone refractory prostate carcinomas. It is possible that amplification facilitates cell growth ability in low concentrations of androgens remaining in the serum after androgen deprivation therapy. We evaluate whether androgen receptor gene amplification at primary progression is associated with response to second line combined androgen blockade for prostate cancer. MATERIALS AND METHODS: A total of 77 patients with prostate cancer were treated initially with androgen deprivation monotherapy followed by combined androgen blockade after the first progression. After initiation of second line combined androgen blockade patients were followed every 3 months to evaluate treatment responses. Biopsies were taken from the prostate at the first progression under endocrine monotherapy. Androgen receptor gene copy number was determined by fluorescence in situ hybridization. RESULTS: Androgen receptor gene amplification was found in 10 of the 77 cases (13%) at the primary disease progression, and was associated with a favorable response to second line combined androgen blockade. Only 1 of 34 (3%) patients classified as nonresponders had androgen receptor gene amplification, whereas 9 of 41 (21%) classified as having either stable disease or response had amplification (p = 0.016). Patients with androgen receptor gene amplification also had a decrease in prostate specific antigen more often after combined androgen blockade than those with no amplification (p = 0.079). However, androgen receptor gene amplification was not associated with patient survival after the first progression. CONCLUSIONS: Androgen receptor gene amplification detected in tumors progressing during androgen deprivation monotherapy is associated with favorable treatment response to second line combined androgen blockade. This finding suggests that at least some androgen receptor amplified tumors retain a high degree of dependency on residual androgens remaining in serum after monotherapy. PMID- 11061899 TI - Molecular markers of cancer progression. Ready or not, here they come. PMID- 11061900 TI - Salvage radical prostatectomy for radiorecurrent prostate cancer: morbidity revisited. AB - PURPOSE: With the advent of prostate specific antigen (PSA) testing and transrectal ultrasound guided prostate biopsy there has been stage migration in the diagnosis of prostate cancer, so that more younger men are being diagnosed with organ confined prostate cancer. Many patients elect radiation therapy, while some have recurrent or new prostate cancer with absent systemic disease and life expectancy greater than 10 years. We present our experience with salvage radical prostatectomy in these cases. MATERIALS AND METHODS: Between 1995 and 2000, 6 men treated with curative intent with radiotherapy for prostate cancer were subsequently treated with salvage surgery for clinically localized prostate cancer. All men had biopsy proved recurrent or persistent prostate cancer, increasing serum PSA, no evidence of systemic disease at surgery and life expectancy greater than 10 years. We assessed the morbidity associated with this procedure and compared results to those in the contemporary literature. RESULTS: Six patients underwent salvage radical prostatectomy. Initial pre-radiation PSA was 4.5 to 15.7 ng./ml. Pre-radiation disease was clinical stage T1c in 5 cases and B2 in 1. The interval from radiotherapy to repeat biopsy was 12 to 48 months. A mean of 6.3 months after local recurrence was detected and before salvage radical prostatectomy was performed 4 patients underwent androgen deprivation therapy. Mean operative time was 195 minutes, intraoperative blood loss was 680 cc, and hospital stay and catheterization time were 3.2 and 13.8 days, respectively. There were no rectal injuries. All 6 patients are impotent, 5 are continent and 1 has mild stress incontinence. There was biochemical failure in 1 case 36 months after salvage radical prostatectomy and no evidence of recurrence in the remaining 5 at a mean followup of 27 months. CONCLUSIONS: Salvage radical prostatectomy is a technically challenging procedure. In the past it was associated with a high incidence of rectal injury, urinary incontinence and anastomotic stricture. The results of our relatively small series are encouraging and concur with those of recent studies that the morbidity of salvage radical prostatectomy is lower than previously reported. We believe that salvage radical prostatectomy may be considered a reasonable treatment option in appropriate patients with radiorecurrent prostate cancer. PMID- 11061901 TI - Laparoscopic suction irrigator for percutaneous removal of renal pelvic bezoar. PMID- 11061902 TI - Fibrin sealant treatment of splenic injury during open and laparoscopic left radical nephrectomy. AB - PURPOSE: We describe the use of fibrin sealant for rapid and definitive hemostasis of splenic injuries incurred during open and laparoscopic left nephrectomy. MATERIALS AND METHODS: In 2 patients undergoing left nephrectomy for a suspicious renal mass splenic laceration occurred during mobilization of the colonic splenic flexure at open nephrectomy and laparoscopic upper pole dissection, respectively. Fibrin sealant was applied topically in each case. RESULTS: In each patient fibrin sealant achieved immediate hemostasis and each recovered without further splenic bleeding. CONCLUSIONS: The topical application of fibrin sealant safely, rapidly and reliably achieves definitive hemostasis of splenic injuries. It is simple to use in the open and laparoscopic approaches. PMID- 11061903 TI - Outcome results of transurethral collagen injection for female stress incontinence: assessment by urinary incontinence score. AB - PURPOSE: We assessed the results of collagen injection for female sphincteric incontinence using strict subjective and objective criteria. MATERIALS AND METHODS: We evaluated 63 consecutive women with sphincteric incontinence who underwent a total of 131 transurethral collagen injections. Sphincteric incontinence was confirmed by urodynamics. All patients were treated with 1 to 5 transurethral collagen injections and treatment outcome was classified according to a new outcome score. Cure was defined as no urinary loss due to urge or stress incontinence documented by a 24-hour diary and pad test, and patient assessment that cure was achieved. Failure was defined as poor objective results and patient assessment that treatment failed. Cases that did not fulfill these cure and failure criteria were considered improved and further classified as a good, fair or poor response. RESULTS: Mean patient age plus or minus standard deviation was 67.7 +/- 12.8 years. All women had a long history of severe stress urinary incontinence, 18 (29%) underwent previous anti-incontinence surgery, and 41% had combined stress and urge incontinence. Preoperatively diary and pad tests revealed a mean of 7.5 +/- 4.6 incontinence episodes and 152 +/- 172 gm. of urine lost per 24 hours. Overall 1 to 5 injections were given in 26, 17, 13, 3 and 4 patients, respectively. Mean interval between injections was 4.4 +/- 5.7 months, mean followup was 12 +/- 9.6 months, and mean interval between the final injection and outcome assessment was 6.4 +/- 4.9 months. There was a statistically significant decrease in the total number of incontinence episodes per 24-hour voiding diary after each injection session. Although there was a clear trend toward decreased urinary loss per 24-hour pad test, statistical significance was not established. Using the strict criteria of our outcome score overall 13% of procedures were classified as cure, 10%, 17% and 42% as good, fair and poor, respectively, and 18% as failure. CONCLUSIONS: As defined by strict subjective and objective criteria, we noted a low short-term cure rate after collagen injection in women with severe sphincteric incontinence. It remains to be determined how patients with less severe incontinence would fare using our outcome assessment instruments. PMID- 11061904 TI - Cortical evoked potentials of the dorsal nerve of the clitoris and female sexual dysfunction in multiple sclerosis. AB - PURPOSE: We evaluated whether disrupting genital central nervous system pathways is associated with subjective reports of sexual dysfunction in women with multiple sclerosis. MATERIALS AND METHODS: We performed pudendal somatosensory evoked potential testing in and had sexual questionnaires completed by 14 women with a mean age of 47 years who had multiple sclerosis. RESULTS: The mean expanded disability status score was 5. All but 1 woman reported the desire for sexual intercourse. There was a high rate of dissatisfaction with their sex life and all study participants had concomitant bladder and bowel function problems. The most common sexual complaint was difficult or no orgasm, which was statistically associated with abnormalities or absence of 1 or both pudendal cortical evoked potentials. Fatigue and arousal disorders were also common. CONCLUSIONS: Women with multiple sclerosis have a high self-reported rate of sexual dysfunction, which decreases quality of life. Electrodiagnostic data imply that pudendal somatosensory innervation is necessary for normal female orgasmic function. More study is needed to confirm these findings. PMID- 11061905 TI - Video urodynamic findings in men with the central cord syndrome. AB - PURPOSE: The central cord syndrome reportedly has a favorable prognosis and rehabilitation outcome. However, to our knowledge the status of the lower urinary tract in patients with the central cord syndrome is unclear. We report on 22 men with the central cord syndrome who were evaluated by video urodynamics. MATERIALS AND METHODS: From 1986 to the present we identified 22 men with a mean age of 51 years who had the central cord syndrome and were included in the Houston Veterans Affairs spinal cord registry. All patients underwent video urodynamic evaluation a mean of 34.5 months after injury. RESULTS: Video urodynamic testing for vesicourethral dysfunction was normal in 3 patients, while it showed bladder outlet obstruction secondary to benign prostatic hyperplasia in 2, detrusor areflexia in 4, external detrusor-sphincter dyssynergia in 11, detrusor hyperreflexia with a synergistic external urethral sphincter in 1 and detrusor hypocontractility in 1. Urinary tract infection recurred in 3 patients with external detrusor-sphincter dyssynergia and urolithiasis developed in 2. CONCLUSIONS: Urodynamic testing revealed a high incidence of external detrusor sphincter dyssynergia in men with the central cord syndrome. Due to the potential for upper tract deterioration all patients with the central cord syndrome should undergo baseline urodynamic studies. Those at high risk for upper tract deterioration with external detrusor-sphincter dyssynergia or a loss of compliance should be treated more aggressively with clean intermittent catheterization and anticholinergic medication when possible. PMID- 11061906 TI - Benign medullary fibroma of the kidney. PMID- 11061907 TI - Hypertension: a rare presentation of renal cell carcinoma. PMID- 11061908 TI - Intrarenal mixed germ cell tumor. PMID- 11061909 TI - Protracted urinary retention necessitating urethrolysis following tension-free vaginal tape surgery. PMID- 11061910 TI - Desquamative penile lesion as a manifestation of Behcet's disease. PMID- 11061911 TI - Testicular metastasis of transitional cell carcinoma of the prostate. PMID- 11061912 TI - Primary leiomyosarcoma of the seminal vesicle. PMID- 11061913 TI - Allopurinol induced meningitis. PMID- 11061914 TI - Re: removal of the entrapped basketed ureteral calculus: a novel technique. PMID- 11061915 TI - Re: Appendicovesicostomy: the Mitrofanoff procedure--a 15-year perspective. PMID- 11061916 TI - Re: A prospective study of interstitial cystitis: results of longitudinal followup of the interstitial cystitis data base cohort. PMID- 11061917 TI - Re: Dermatomyositis associated with testicular germ cell cancer. PMID- 11061918 TI - Re: Radical prostatectomy for localized prostate cancer provides durable cancer control with excellent quality of life: a structured debate. PMID- 11061919 TI - Re: Prostate specific antigen bounce after radioactive seed implantation followed by external beam radiation for prostate cancer. PMID- 11061920 TI - Magnetic resonance imaging of the dysplastic renal moiety and ectopic ureter. AB - PURPOSE: We determined the role of magnetic resonance imaging (MRI) in symptomatic children with clinically suspected and radiologically occult dysplastic renal moieties and ectopic ureters. MATERIALS AND METHODS: We reviewed clinical, imaging, cystoscopic, surgical and histological findings in 6 symptomatic children 1 to 15 years old with dysplastic renal moieties. RESULTS: After multiple conventional imaging studies failed to delineate urinary tract anatomy MRI provided detailed multiplanar images of dysplastic renal moieties that were diagnostic and predictive of subsequent intraoperative findings. Dysplastic upper pole moieties identified in 4 children were associated with ectopic ureters inserting into the vagina, prostatic urethra, bladder neck and bladder neck ureterocele in each. A solitary kidney with contralateral blind ending ectopic ureters inserted into the bladder base in 2 cases. Pelvic cystic structures visualized by ultrasound in 3 patients were tortuous distal ureters on MRI. MRI specifically identified ureteral insertion sites that were not evident in 3 of the 5 patients who underwent cystoscopy. CONCLUSIONS: MRI may facilitate diagnosis, guide cystoscopy and aid in preoperative planning in children with poorly functioning renal moieties and ectopic ureters. PMID- 11061921 TI - Ectopic ureterocele: results of open surgical therapy in 40 patients. AB - PURPOSE: The treatment of ectopic ureterocele is controversial. In addition to debate on optimal therapy, discussion exists on whether there is further risk of deteriorating bladder function after extensive bladder surgery during the first year of life, which is a reason to postpone surgery. In a prospective nonrandomized trial we treated 40 patients regardless of age who had ectopic ureterocele with complete surgical reconstruction of the lower urinary tract and upper pole resection of poorly functioning upper pole moieties at referral. Excluded from study were 3 patients with only 1 affected renal moiety initially. MATERIALS AND METHODS: We treated 31 female and 9 male patients 0 to 8.8 years old (mean age 2.17) at surgery for ectopic ureterocele extending into the bladder neck and urethra, including 19 younger than 1 year. Primary ureterocele excision was performed in 37 cases with reconstruction of the urethra, bladder neck and trigone, and ureteral reimplantation. Because of small ureterocele size, the ureterocele was left in situ in 3 patients, leading to secondary ureterocele removal due to obstructive voiding and urinary incontinence in 1 each. A staged procedure in 5 neonates involved primary lower urinary tract reconstruction with upper pole cutaneous ureterostomies followed by upper pole resection or ureteral reimplantation a few months later. After bladder neck reconstruction in 16 cases colposuspension was also done to create a normal vesicourethral angle. All patients underwent clinical and urodynamic evaluation at least 1.25 years after surgery (mean followup 5.59). Patients who were too young for the clinical assessment of continence by January 1999 were excluded from study. RESULTS: All patients are continent. A secondary endoscopic procedure was required in 13 cases, including cystoscopy only in 2, scar incision near the ureteral orifice in 3, endoscopic reflux treatment in 4, ureterocele remnant resection in 2 and bladder neck incision for obstructive voiding in 2. Secondary open bladder reconstruction was performed in another case due to a diverticulum. Postoperatively only 1 or 2 uncomplicated episodes of urinary tract infection developed in 11 patients, while there were recurrent urinary tract infections in 4. In a patient with a preexisting loss of renal function a severe infection led to renal scarring. The voiding pattern was normal in 29 patients but 11 had dysfunctional voiding, including 5 with recurrent urinary tract infection. Urodynamic followup confirmed these clinical findings. Bladder capacity in these patients was relatively high at an average of 124% of expected capacity for age. We noted no statistically significant difference in followup parameters in patients who underwent surgery before and after age 1 year. Additional colposuspension in 16 patients did not result in any significant change in outcome compared with that in patients without this procedure. CONCLUSIONS: When compared with results in the literature, complete primary lower urinary tract reconstruction in patients with ectopic ureterocele appears to have better results than a staged approach with initial endoscopic treatment. Moreover, our study provides no proof that extensive reconstructive bladder surgery in neonates and infants leads to bladder function deterioration at a later age. PMID- 11061922 TI - Further experience with seromuscular colocystoplasty lined with urothelium. AB - PURPOSE: We report our continuing experience with seromuscular colocystoplasty lined with urothelium. This procedure is designed to preserve the urothelium and potentially decrease the incidence of complications associated with standard bladder augmentation. MATERIALS AND METHODS: We retrospectively reviewed the charts of 32 patients who underwent seromuscular colocystoplasty lined with urothelium between April 1994 and July 1999. Data were collected on patient demographics, surgical indications, previous and adjunctive surgical procedures, preoperative and postoperative urinary continence, upper urinary tract changes, urodynamic parameters, surgical complications and histological findings. RESULTS: Mean patient age at surgery plus or minus standard deviation was 11.1 +/- 4.8 years. Mean followup was 1.6 +/- 1 years. A mean of 1.5 +/- 0.9 years postoperatively urodynamic studies available in 28 cases showed that total and safe bladder capacity increased by 1.8 and 2.4-fold, respectively. Continence was achieved in 71% of patients after the initial procedure, increasing to 81% after secondary procedures. Hourglass deformity developed in 7 cases (22%), augmentation failed in 4 (12.5%) and there were bladder calculi in 2 (6%). New onset or increased hydronephrosis and reflux were present in 6 of 62 (10%) and 9 of 60 (15%) evaluated renal units, respectively. Of the 7 interpretable biopsies 5 revealed various degrees of repeat colonic mucosal growth. There was no bladder perforation or metabolic abnormalities, and mucous production was not clinically significant. CONCLUSIONS: Seromuscular colocystoplasty lined with urothelium is a viable alternative to standard bladder augmentation. The 2 procedures have a similar overall complication rate. Comparatively there appears to be a low incidence of bladder calculi, mucous production has not been clinically significant, metabolic disturbances have not developed and perforation has not occurred during short-term followup. We are enthusiastic about this technique and continue to apply it in select patients. PMID- 11061923 TI - Voiding pattern in healthy children 0 to 3 years old: a longitudinal study. AB - PURPOSE: We describe the development of voiding patterns and bladder control in healthy children during the first 3 years of life. MATERIALS AND METHODS: We determined voiding patterns, bladder capacity and post-void residual urine volume per 4 hours individually and noninvasively every 3 months in 36 female and 23 male healthy infants using the 4-hour voiding observation. RESULTS: Voiding frequency decreased slowly from 5 to 2 voiding episodes per 4 hours from ages 3 months to 3 years. We noted interrupted voiding in 33% of subjects at age 3 months but this condition was rare after age 2 years. Voiding during sleep occurred mainly during the first 7 months of life and did not continue after age 18 months. Bladder capacity increased from a median of 52 to 67, 68 and 123 ml. during years 1 to 3, respectively. As measured by post-void residual urine volume, bladder emptying was unchanged during years 1 and 2 but it decreased during year 3 (median 6 versus 0 and mean 4 versus 3 ml. per 4 hours). CONCLUSIONS: During the first 3 years of life the number of voiding episodes, including interrupted voiding, post-void residual urine and voiding during sleep, decreased while bladder capacity increased. PMID- 11061924 TI - Results of umbilicoplasty for bladder exstrophy. AB - PURPOSE: The umbilicus is an important aesthetic landmark and its absence or deformity may be associated with poor self-image. In patients born with bladder exstrophy the umbilicus is attached to the upper margin of the bladder and reconstructive surgery often removes the navel. The umbilicus marks the waistline and serves to complete the harmony of the curved lines above and below the waist. We present our experience with children born with exstrophic anomalies during the last 2 decades. MATERIALS AND METHODS: Our database included 61 children born with classic bladder and 8 born with cloacal exstrophy treated between 1980 and 1998. We performed primary reconstruction in 35 children, while 34 children and young adults were referred for secondary surgical repair, including bladder augmentation, continent diversion, genitoplasty and so forth. Neoumbilicoplasty was done in all of the former and in 30 of the 34 latter cases. Early in the series a V-shaped flap was raised and buried subcutaneously. The flap eventually became a tube around the cystotomy tube and the cicatrix formed the umbilical dimple. This method necessitated packing with iodoform gauze for 4 weeks with weekly dressing. The technique evolved into a tubularized U-shaped flap. A rubber tube was placed indwelling as a stent to maintain inward projection of the neoumbilicus. RESULTS: In 66 of the 69 cases the early results of umbilicoplasty were described by the surgeon as excellent or satisfactory. In 3 cases the neoumbilicus appeared flat, lost depth and was described as unsatisfactory. Long term followup of more than 1 year was available in 48 patients, of whom 2 underwent umbilical repositioning for an off center or low umbilicus and 3 underwent repeat umbilicoplasty for a flat umbilicus that had lost depth. The best cosmetic results were achieved in patients with a relatively thick layer of subcutaneous fat, whereas cosmesis was suboptimal in thin children. Nevertheless, the patients and parents were generally pleased with the umbilical appearance even when the surgeon was not. CONCLUSIONS: Although the navel is a functionless depressed scar, it represents an important and pleasing landmark. Umbilical construction should be attempted early during functional closure or urinary diversion. PMID- 11061925 TI - Intraoperative pharmacological erection as an aid to pediatric hypospadias repair. AB - PURPOSE: The intraoperative evaluation of erection is a major advance in hypospadias surgery. We determined the advantages of erections induced pharmacologically over those induced by intracorporeal saline injection for evaluating chordee during hypospadias surgery. MATERIALS AND METHODS: During the repair of hypospadias or chordee without hypospadias 56 boys 6 months to 13 years old underwent pharmacological erection induced by 14 microg. alprostadil administered intracavernously. Phenylephrine (40 microg. ) was given for detumescence. We monitored the adequacy of erection and detumescence, changes in blood pressure and pulse, and in 3 cases intracorporeal pressure. Intraoperative artificial erection was also induced in 30 patients. RESULTS: Erection occurred within 1 minute of injection. It was judged to be excellent in 47 cases and adequate in 6, while it failed in 3 probably due to injection outside of the corpora. Erection involved the whole penis, in contrast to artificial erection when tourniquet placement altered the erection and left the penile base flaccid. The degree of chordee remained stable during evaluation compared to artificial erection when curvature varied with the force of the saline injection. Erection persisted during chordee repair as long as the corpora were not opened. Detumescence occurred within seconds in all cases in which phenylephrine was given. There were no cases of priapism, and systemic blood pressure and pulse did not change. Intracorporeal pressure during pharmacological erection was 47 to 70 mm. Hg, whereas during artificial erection pressure was 50 to 250 mm. Hg depending on how much saline was injected and how rapidly fluid drained through the tourniquet. Chordee was induced by over injection. CONCLUSIONS: Pharmacological erection in hypospadias repair is effective and reliable with no significant complications. It is especially valuable in severe hypospadias and in patients with a large suprapubic fat pad. Artificial erection with saline injection should be performed with only moderate force since over filling is unphysiological and may falsely induce chordee. PMID- 11061926 TI - Periurethral muscle complex reassembly for exstrophy-epispadias repair. AB - PURPOSE: Continence is a difficult goal in exstrophy-epispadias complex repair. It is presumed that all anatomical components involved in the exstrophy epispadias abnormality are present but laterally and anteriorly displaced. The penile disassembly technique for epispadias restores the normal anatomical relationship of the male genital components. Its extension to complete primary bladder exstrophy closure enables deeper positioning of the bladder neck within the pelvic diaphragm. We identified the perineal striated muscular complex and present its appropriate periurethral reassembly as a main step in exstrophy epispadias complex repair. MATERIALS AND METHODS: Bladder exstrophy and epispadias repairs were performed in 10 male and 3 female consecutive patients with the exstrophy-epispadias complex, including 1-stage reconstruction in 2 male newborns and 2 females with exstrophy, and as further surgery in a female with cloacal exstrophy and previous failed 1-stage repair, 4 males with incontinent epispadias (secondary repair in 1) and 4 males with epispadias in whom exstrophy closure had been previously done. In the males after bladder plate closure and corporeal body splitting a sagittal incision was made in the intersymphyseal tissue and extended posteriorly to the perineal body midline. The bipolar electrical stimulator was used to identify pelvic muscle components in the sagittal plane and reapproximate them along the tubularized posterior urethra to form the periurethral muscle complex. In the 3 females the urethral plate and vagina were similarly mobilized posterior through the sagittal incision of the perineal body. No patient underwent bladder neck plasty. RESULTS: At 9 months to 4 years of followup cosmesis was good in 12 patients, while 1 required secondary glanular urethroplasty. There was mild pyelectasis in 3 cases but no severe hydronephrosis and no renal function deterioration. Pyelonephritis developed in 6 patients (46%). Cystography at 1 year showed that bladder capacity was 35 to 80 and 65 to 120 cc in exstrophy and epispadias cases, respectively. There was cyclic voiding with 30 to 90-minute dry intervals in 7 patients (54%), of whom 5 had exstrophy and 2 had epispadias. Daytime voiding control with a 2 to 3-hour voiding interval was achieved in 1 female with exstrophy and 2 patients with epispadias (23%). Incontinence was present in 2 patients with previous exstrophy closure and 1 with cloacal exstrophy (23%). CONCLUSIONS: Early restoration of a physiological vesicourethral balance of coordinated activity is feasible for the progressive achievement of continence in patients with the exstrophy-epispadias complex. Sagittal splitting of the perineal tissue with identification of the muscle components as well as midline reassembly of the periurethral striated muscular complex helps to reconfigure the pelvic anatomy in a more normal fashion and allows better restoration of coordinated vesicourethral activity. PMID- 11061927 TI - Holmium laser lithotripsy for ejaculatory duct calculi. PMID- 11061928 TI - Testicular immature teratoma with primitive neuroectodermal tumor in early childhood. PMID- 11061929 TI - Does the urorectal septum fuse with the cloacal membrane? AB - PURPOSE: Traditional theories of cloacal embryogenesis assume that the urorectal septum fuses with the cloacal membrane before the anal membrane disintegrates. However, recent observations in humans and other species raise doubt about this assumption. We determined whether urorectal septum fusion occurs in rats. MATERIALS AND METHODS: Rat embryos were harvested at specific times between days 11 and 16 of gestation. We evaluated the morphology, growth and relationship of the urorectal septum to the cloacal membrane on serial histological sections. RESULTS: The urorectal septum consistently fused with the cloacal membrane on day 15 of gestation before the cloacal membrane began to disintegrate. CONCLUSIONS: In rats the urorectal septum fuses with the cloacal membrane, after which the urogenital membrane and anal membrane disintegrate by a process of apoptosis. PMID- 11061930 TI - Pediatric oncology refracted through the prism of Wilms tumor: a discourse. PMID- 11061931 TI - Robotic remote laparoscopic nephrectomy and adrenalectomy: the initial experience. AB - PURPOSE: We evaluated the feasibility of performing laparoscopic nephrectomy and adrenalectomy exclusively by using robotic telepresent technology from a remote workstation and compared outcomes with those of conventional laparoscopy in an acute porcine model. MATERIALS AND METHODS: Five pigs underwent bilateral laparoscopic nephrectomy (robotic in 5 and conventional in 4) and adrenalectomy (robotic in 4 and conventional in 3). In the 9 robotic laparoscopic procedures all intraoperative manipulations were completely performed telerobotically from a remote workstation without any conventional laparoscopic assistance on site. Animals were sacrificed acutely. RESULTS: Robotic laparoscopic nephrectomy required significantly longer total operative (85.2 versus 38.5 minutes, p = 0.0009) and actual surgical (73.4 versus 27.5 minutes, p = 0.0002) time than conventional laparoscopy. However, blood loss and adequacy of surgical dissection were comparable in the 2 groups. Robotic laparoscopic adrenalectomy required longer total operative (51 versus 32.3 minutes, p = 0.13) and actual surgical (38.5 versus 18.7 minutes, p = 0.14) time than conventional laparoscopy. The solitary complication in this study was an inferior vena caval tear during robotic right adrenalectomy, which was adequately repaired by sutures telerobotically in a remote manner. CONCLUSIONS: To our knowledge we present the initial experience with remote telerobotic laparoscopic nephrectomy and adrenalectomy. Telepresent laparoscopic surgery is feasible. PMID- 11061932 TI - Feasibility of pathological evaluation of morcellated kidneys after radical nephrectomy. AB - PURPOSE: The indications for laparoscopic nephrectomy have grown to include renal malignancy. Although morcellation of these specimens has been described, to our knowledge we present the first systematic review of the feasibility and validity of pathological evaluation of these tumors with regard to grade and stage. MATERIALS AND METHODS: Nine formalin fixed and 5 fresh intact radical nephrectomy specimens were evaluated by 2 pathologists before and after high speed electrical tissue morcellation. The ability to distinguish tissue histology, and tumor size, stage and grade were compared. Impermeability of the laparoscopy sack after morcellation was also evaluated using indigo carmine stained normal saline placed in the used sack. RESULTS: The 9 preserved specimens included 7 renal cell carcinomas and 2 oncocytomas, while 4 of the 5 fresh specimens were renal cell carcinoma and 1 was oncocytoma. Overall tumor size was 2 to 7 cm. (mean 4.9). The 4 fresh renal cell carcinomas were of the clear cell type. Comparison of pathological evaluation after morcellation by another pathologist revealed identical histology, grade and stage for each tumor. Four cases of perinephric fat invasion (3 fixed and 1 fresh specimens) were identified after morcellation. Only tumor size was not assessed after morcellation. Laparoscopy sack integrity was confirmed in 13 of 14 cases. In 1 case involving a formalin fixed specimen a gross defect in the laparoscopy sack was demonstrated after morcellation. CONCLUSIONS: Morcellation of radical nephrectomy specimens in vitro did not alter the determination of histology, grade or local invasiveness of tumor. For all fresh tissue and remarkably for all but 1 formalin fixed tissue specimen the laparoscopy sack remained intact. Preliminary data from this in vitro model imply that limited in vivo morcellation of radical nephrectomy specimens may be performed without sacrificing staging information. PMID- 11061933 TI - Expression of the SART3 tumor rejection antigen in renal cell carcinoma. AB - PURPOSE: We recently reported that SART3 tumor rejection antigen is recognized by HLA class I restricted cytotoxic T lymphocytes from patients with esophageal cancer. We now investigate the expression of SART3 antigen in renal cell carcinoma to identify an appropriate molecule that may be used in specific immunotherapy of renal cell carcinoma. MATERIALS AND METHODS: Renal cell carcinoma and nontumorous kidney tissues were obtained at surgery. A section of each sample was minced with scissors and stored at -80C until use. SART3 antigen expression was examined in uncultured renal cell carcinoma and nontumorous kidney tissues. We also evaluated the ability of derived peptides to include cytotoxic T lymphocytes in peripheral blood mononuclear cells from patients with renal cell carcinoma. RESULTS: The SART3 antigen was detected in all renal cell carcinoma cell lines, primary cultures of renal cell carcinoma and nontumorous kidney tissues, and in the cytosol of 57% and 15% of renal cell carcinoma and nontumorous kidney tissues, respectively. HLA-A2402 restricted and tumor specific cytotoxic T lymphocytes (KE4) used in cloning of the SART3 gene were significantly cytotoxic to cells from renal cell carcinoma cell lines and primary cultures of renal cell carcinoma tissue but they did not lyse normal cells, including those from primary cultures of nontumorous kidney tissue. The SART3 peptides derived from positions 109-118 and 315-323 induced HLA-A24 restricted cytotoxic T lymphocytes to renal cell carcinoma cells from peripheral blood mononuclear cells of patients with renal cell carcinoma. CONCLUSIONS: The SART3 antigen and derived peptides may be applied to the specific immunotherapy of HLA A24+ renal cell carcinoma. PMID- 11061934 TI - The effect of pneumoperitoneum on dissemination and scar implantation of intra abdominal tumor cells. AB - PURPOSE: The role of laparoscopy for the treatment of cancer remains controversial, and a particular concern is port site metastases after laparoscopic surgery. Since laparoscopy is being performed with increasing frequency, the question arises as to whether it is a safe oncological procedure. After intraperitoneal inoculation of renal cell carcinoma cells in a mouse model, we compare abdominal wall scar implantation following laparoscopic trocar insertion and pneumoperitoneum with standard laparotomy, and examine the effects on tumor dissemination in the peritoneal cavity. MATERIALS AND METHODS: Following intra-abdominal RENCA cell inoculation, Balb/c mice were randomized into group 1 20 mice that underwent carbon dioxide pneumoperitoneum and telescope trocar insertion, group 2-20 subjected to laparotomy and group 3-10 anesthetized only. All animals were sacrificed 2 weeks after inoculation, and abdominal wall metastases and intraperitoneal tumor distribution were evaluated. RESULTS: Overall, intra-abdominal implantation of inoculated RENCA tumor cells was detected in 15 of 20 animals (75%) in group 1, 14 of 20 (70%) in group 2 and 10 of 10 (100%) in group 3. Wound metastases developed in 46.7% of the mice in group 1 and 50% in group 2. CONCLUSIONS: There was no difference among the groups in the pattern of intraperitoneal tumor implants and scar seeding incidence. Pneumoperitoneum does not facilitate port site metastases. PMID- 11061935 TI - Flow characteristics of 3 unique ureteral stents: investigation of a Poiseuille flow pattern. AB - PURPOSE: The pattern of flow in the stented ureter (intraluminal and/or extra luminal) has only been defined for the standard pigtail stent. No data are available on stent flow for any specialty stents. To our knowledge we present the first investigation characterizing the type of flow through a stent (Poiseuille versus nonPoiseuille flow). MATERIALS AND METHODS: Flow was measured in an unstented ureter, a standard 7Fr double pigtail stent and the 7/3Fr Tail stent, 7Fr Spirastent and 14/7Fr endopyelotomy stent using a previously developed stent flow model. In vitro pressure flow studies were also done in nonfenestrated 14/7Fr, Tail and standard 7Fr stents. These stents were infused at a constant flow rate of 2 to 10 ml. per minute with monitoring of the corresponding pressure gradients. Resistance to flow was determined for these stents using pressure flow plots and Poiseuille's law. RESULTS: In vivo the 7Fr pigtail, 14/7Fr endopyelotomy and 7/3Fr Tail stents had statistically similar flow rates. Flow through each of these stents exceeded the flow through an unstented ureter. The Spirastent had the least flow in all categories tested. There was no correlation of Poiseuille flow parameters measured in vitro for nonfenestrated stents with in vivo stent flow. CONCLUSIONS: In stented ureters fluid drains through and around the stent regardless of its design. The flow characteristics of these 3 specialty stents were not predictable according to lumen or stent size. In vitro Poiseuille's flow did not correlate with in vivo stent flow. PMID- 11061936 TI - Bacterial adherence in a rat bladder augmentation model: ileocystoplasty versus colocystoplasty. AB - PURPOSE: Various intestinal segments are used to reconstruct the urinary tract. For unclear reasons asymptomatic chronic bacteriuria is common in patients treated with reconstruction. We compared bacterial adherence in ileum, colon and bladder in rats with ileal and colonic bladder augmentation. MATERIALS AND METHODS: Bladder augmentation using ileum or colon was performed in 8-week-old rats. After 3 months urinary pH was measured and urine was cultured. Urovirulence factors of Escherichia coli aspirated from the augmented bladders were detected by polymerase chain reaction. In rats with negative urine culture after augmentation experimental cystitis was induced by the transurethral inoculation of E. coli C5, with type I pili and aerobactin or E. coli C92 with type I pili, P fimbriae and aerobactin at a concentration of 10(5) colony forming units per 0.3 ml. After 14 days we counted the colony forming units per cm.(2) of bladder and cm.(2) of intestinal augmentation tissue. RESULTS: When cultures were negative, mean urinary pH plus or minus standard deviation for ileocystoplasty (7.35 +/- 0.33) was significantly higher than that for colocystoplasty (6.80 +/- 0.45) or in controls (6.67 +/- 0.30). Bacterial colonization occurred in 60 of 96 ileocystoplasties (62.5%) and 36 of 68 colocystoplasties (52.9%). All 32 E. coli strains aspirated from ileocystoplasties had type I pili. In colocystoplasties 14 strains had type I pili, 4 had P fimbriae and type I pili, and 1 had no virulence factor. In experimental cystitis in the ileal patch and bladder there were 10(3.2) to 10(6.2) (log mean 4.9) and 10(1.1) to 10(5.1) (log mean 3.5) colony forming units of E. coli C5, respectively. In the colonic patch and bladder there were 10(2.2) to 10(6.2) (log mean 3.9) and 10(2.1) to 10(5.1) (log mean 3.7) colony forming units of E. coli C5, respectively. In the ileal patch and bladder versus the colonic patch and bladder there were 10(3.2) to 10(6.2) (log mean 5.0) and 10(3.1) to 10(6.1) (log mean 4.5) versus 10(3.2) to 10(6.2) (log mean 4.3) and 10(2.1) to 10(6.1) (log mean 3.8) colony forming units of E. coli C92, respectively. E. coli C5 adhered to more ileum than bladder, while bacterial adherence did not differ for colon and bladder. Adherence of E. coli C92 did not differ significantly in bladder and implanted ileum or colon. CONCLUSIONS: The colonic segment offers more resistance to E. coli than the ileal segment in urinary diversion. PMID- 11061937 TI - Electromotive administration of intravesical bethanechol and the clinical impact on acontractile detrusor management: introduction of a new test. AB - PURPOSE: It is often difficult to determine the functional status of the detrusor muscle in patients with detrusor areflexia. We performed a clinical study to establish a test defining residual detrusor capacity in such patients. MATERIALS AND METHODS: In phase 1, 5 controls with detrusor areflexia were tested with an intravesical instillation of 20 mg. bethanechol in 150 cc of sodium chloride 0.3% with and without 20 mA. of pulsed current applied via an electrode catheter through the saline. Cystometry simultaneously recorded intravesical pressure changes. In phase 2, 45 patients with detrusor areflexia were tested with electromotive administration of intravesical bethanechol. In phase 3, 25 mg. bethanechol given orally once daily were prescribed for 15 patients and voiding control was assessed after 6 weeks of therapy. RESULTS: Neither bethanechol without current nor current through saline only led to increased intravesical pressure. However, we noted a mean pressure increase of 34 cm. water during the electromotive administration of bethanechol in 24 of 26 patients with areflexia and neurological disease compared to only 3 cm. water in 3 of 11 with a history of chronic bladder dilatation. Oral bethanechol restored spontaneous voiding in 9 of 11 patients who had had a positive response to the electromotive administration of bethanechol, whereas all 4 without a pressure increase during the electromotive administration of bethanechol did not void spontaneously. CONCLUSIONS: Electromotive administration of intravesical bethanechol identifies patients with an atonic bladder and adequate residual detrusor muscle function who are candidates for restorative measures, such as oral bethanechol and intravesical electrostimulation. Those who do not respond to the electromotive administration of bethanechol do not benefit from oral bethanechol and are candidates for catheterization. PMID- 11061938 TI - Bladder epithelial cells from patients with interstitial cystitis produce an inhibitor of heparin-binding epidermal growth factor-like growth factor production. AB - PURPOSE: The etiology of interstitial cystitis is unknown. We previously identified an interstitial cystitis urine factor, antiproliferative factor, that inhibits proliferation of bladder epithelial cells in vitro and complex changes in epithelial growth factor levels, including profound decreases in heparin binding epidermal growth factor-like growth factor (HB-EGF). Bladder and renal pelvic catheterization of patients with interstitial cystitis indicated that the antiproliferative factor is made and/or activated in the distal ureter or bladder. Therefore, we determined whether bladder epithelial cells from interstitial cystitis cases produced the antiproliferative factor and whether purified antiproliferative factor could alter production of growth factors known to be abnormal in interstitial cystitis. MATERIALS AND METHODS: Antiproliferative factor activity was determined by 3H-thymidine incorporation into primary bladder epithelial cells. The antiproliferative factor was purified by size fractionation followed by sequential chromatography involving ion exchange, hydrophobic interaction and high performance liquid chromatography. HB-EGF, epidermal growth factor, insulin-like growth factor and insulin-like growth factor binding protein 3 levels were determined by enzyme-linked immunosorbent assay. RESULTS: Bladder epithelial cells from patients with interstitial cystitis produced a single antiproliferative factor with the same purification profile as that purified from interstitial cystitis urine. Purified antiproliferative factor specifically inhibited HB-EGF production by bladder epithelial cells in vitro, and the effect of interstitial cystitis urine or purified antiproliferative factor on bladder cell proliferation was inhibited by recombinant human HB-EGF in a dose dependent manner. Similar to urine HB-EGF, serum HB-EGF was also significantly lower in interstitial cystitis cases than in controls. CONCLUSIONS: Bladder epithelial abnormalities in interstitial cystitis may be caused by a negative autocrine growth factor that inhibits cell proliferation by down-regulating HB-EGF production. Furthermore, decreased levels of urine and serum HB-EGF indicate that interstitial cystitis may be a urinary tract manifestation of a systemic disorder. PMID- 11061939 TI - Pentosanpolysulfate inhibits mast cell histamine secretion and intracellular calcium ion levels: an alternative explanation of its beneficial effect in interstitial cystitis. AB - PURPOSE: Mast cells are ubiquitous cells derived from the bone marrow and are responsible for allergic reactions as they release numerous vasodilatory, nociceptive and pro-inflammatory molecules in response to immunoglobulin E (IgE) and specific antigen. Mast cell secretion is also triggered by a number of peptides, such as bradykinin and substance P, and may also be involved in the development of inflammatory responses. An example is interstitial cystitis, which is a sterile painful bladder disorder that has been associated with a defective glycosaminoglycan bladder mucosal layer and an increased number of activated mast cells. Pentosanpolysulfate is a synthetic, sulfated polysaccharide that has been approved for the treatment of interstitial cystitis on the premise that it may replenish the defective glycosaminoglycan layer. We hypothesize that pentosanpolysulfate may also have an additional or alternate action on bladder mast cells. We report that pentosanpolysulfate has a powerful dose dependent inhibitory effect on mast cell release of histamine induced by the mast cell secretagogue compound 48/80. MATERIALS AND METHODS: Inhibition of mast cell secretion was documented by light and electron microscopy and extended to stimulation by substance P or IgE and antigen. RESULTS: The inhibition was more potent than that seen with the clinically available mast cell stabilizer disodium cromoglycate (cromolyn). Maximal inhibition by pentosanpolysulfate was apparent within 1 minute, was unaffected by the length of pre-incubation and persisted after the drug was washed off. In contrast, the effect of cromolyn was limited by rapid tachyphylaxis. In addition, while cromolyn has no effect on mucosal or rat basophilic leukemia cells, pentosanpolysulfate inhibited histamine secretion from both. Confocal microscopy using a calcium indicator dye showed that pentosanpolysulfate decreased intracellular calcium ion levels. CONCLUSIONS: Pentosanpolysulfate appears to be a potent inhibitor of allergic and nonimmune mast cell stimulation, which is an alternative explanation of its benefit in interstitial cystitis. PMID- 11061940 TI - Plasma glutathione S-transferase pi 1-1 AND alpha 1-1 levels in patients with bladder cancer. AB - PURPOSE: Transitional cell carcinomas of the human bladder and many gastrointestinal tumors often contain high amounts of the detoxification enzyme glutathione S-transferase pi (GSTP1-1). Elevated levels of GSTP1-1 have been found in serum and plasma from patients with gastrointestinal, lung or head and neck cancer. GSTP1-1 and glutathione S-transferase alpha (GSTA1-1) have been reported to be increased in 10 of 15 patients (67%) with bladder cancer. We evaluate the role of GSTP1-1 and GSTA1-1 as plasma tumor markers in 50 patients with bladder cancer before and after treatment. MATERIALS AND METHODS: Blood from patients with bladder cancer was sampled in ethylenediaminetetraacetic acid tubes. Plasma GSTA1-1 and GSTP1-1 were measured using the sensitive and specific sandwich enzyme-linked immunosorbent assay. RESULTS: Respective plasma GSTA1-1 and GSTP1-1 levels were above the upper normal reference limit in 2 (4%) and 14 (28%) of the 50 patients with bladder cancer. No significant decrease in plasma GSTA1-1 or GSTP1-1 was noted in matched pairs of plasma samples collected before and after treatment. CONCLUSIONS: In contrast to earlier reports, only a limited number of patients with bladder cancer had increased plasma GSTA1-1 or GSTP1-1, which did not decrease after tumor resection. These findings argue against the use of GSTP1-1 or GSTA1-1 as plasma markers for bladder cancer. PMID- 11061941 TI - Urinary Interleukin-8 and 18 predict the response of superficial bladder cancer to intravesical therapy with bacillus Calmette-Guerin. AB - PURPOSE: We evaluate the predictive value of urinary cytokine levels of interleukin (IL) 8 and 18 for response in patients receiving intravesical bacillus Calmette-Guerin (BCG) for prevention of recurrences of superficial bladder cancer and treatment of carcinoma in situ. MATERIALS AND METHODS: In 28 patients with superficial bladder cancer treated with BCG IL-8 expression in the urine during the first 6 hours after the first BCG instillation was determined. In 17 patients IL-18 levels were also evaluated during the first 12 hours after BCG instillation. IL-8 and 18 levels were determined by solid phase double ligand enzyme-linked immunosorbent assay. RESULTS: In 12 of the 28 patients assessed for IL-8 expression disease recurred after a median followup of 66 months. Median IL 8 expression during the first 6 hours for these patients was 851 ng. (range 232 to 8,497). Median IL-8 expression during the first 6 hours in patients without recurrence was 4,200 ng. (range 432 to 12, 232). Of 8 patients with a followup of greater than 36 months 7 (88%) had no recurrent disease and IL-8 levels greater than 4,000 ng. Patients secreting more than 4,000 ng. IL-8 into the urine after BCG have a significantly higher chance of remaining disease-free (p <0.05), and those with elevated IL-18 expression have a significantly longer disease-free survival (p <0.05). After a median followup of 23 months (range 7 to 93) 6 of the 17 patients assessed for IL-18 expression had treatment failure. Median IL-18 expression in those patients during the first 12 hours was 2,632 pg. (range 860 to 8,298). Median IL-18 expression during the first 12 hours in patients without recurrence was 12,258 pg. (range 1,727 to 151,495). CONCLUSIONS: In this study we confirmed the value of quantitative IL-8 expression in the urine during the first 6 hours after BCG instillation for superficial bladder cancer to predict freedom of disease. Furthermore, to our knowledge we report for the first time the potential value of IL-18 expression in the urine during the first 12 hours after BCG to predict freedom from disease. These findings may help improve the treatment of patients with superficial bladder cancer, especially by identifying those with a high risk of disease recurrence and progression after BCG therapy. PMID- 11061942 TI - Identification by quantitative chromatin pattern analysis of patients at risk for recurrence of superficial transitional bladder carcinoma. AB - PURPOSE: Based on the actual clinical outcomes of 132 fully documented patients with superficial transitional cell carcinoma of the bladder, we characterize the risk of recurrence and/or progression by computer assisted image microscopy applied to Feulgen stained nuclei. MATERIALS AND METHODS: Each tumor was characterized by the conventional grading and staging systems as well as by cytometry generated variables describing nuclear DNA content, nuclear morphometry and chromatin patterns. These data were submitted to discriminant analysis to establish a model distinguishing between 2 groups of patients. Group 1 included cases with remission for more than 60 months and group 2 cases presented with recurrence with or without progression within 12 months of transurethral bladder resection. This latter model was then validated by Kaplan-Meyer analysis of the full data set. RESULTS: As evidenced by Kaplan-Meier analysis, the discriminant factor generated by discriminant analysis of cytometry generated variables provided a cutoff value for distinguishing between low and high risks of recurrence (p <0.00001). In contrast, conventional grading and staging systems were not able to make such efficient distinction. CONCLUSIONS: These 2 groups can be used as references with which new cases can be compared to prognosticate disease behavior independently of histopathological grading and/or clinical staging. PMID- 11061943 TI - Possible role of human growth hormone in penile erection. AB - PURPOSE: Treatment with recombinant human growth hormone in adult patients with growth hormone deficiency increases nitric oxide and cyclic guanosine monophosphate (cGMP). We examined the functional in vitro effects of recombinant human growth hormone on tissue tension and cyclic nucleotide levels of human corpus cavernosum and detected changes in growth hormone in the cavernous and peripheral blood during different phases of penile erection. MATERIALS AND METHODS: Relaxant responses of human corpus cavernosum were investigated using the organ bath technique. Tissue levels of cGMP were determined by a specific radioimmunoassay after dose dependent exposition of isolated human corpus cavernosum strips to recombinant human growth hormone. In 35 healthy potent volunteers blood samples were obtained simultaneously from the corpus cavernosum and cubital vein during different functional conditions of the penis, including flaccidity, tumescence, rigidity and detumescence. Penile erection was induced by audiovisual and tactile stimulation. Serum growth hormone was determined by an immunoradiometric assay. RESULTS: Recombinant human growth hormone elicited dose dependent relaxation of human corpus cavernosum strips in vitro. The relaxing potency of recombinant human growth hormone was paralleled by its ability to elevate intracellular levels of cGMP. In vivo the peripheral growth hormone serum profile of the respective penile conditions did not significantly differ from those of cavernous serum. The main increase in growth hormone to greater than 90% was determined during developing penile tumescence, followed by a transient decrease afterward. CONCLUSIONS: These results suggest that penile erection may probably be induced by growth hormone through its cGMP stimulating activity on human corpus cavernosum smooth muscle. PMID- 11061944 TI - Sorting the hype from the facts in testicular cancer: is testicular cancer related to trauma? AB - PURPOSE: The rate of testicular cancer is increasing. Trauma severe enough to cause testicular atrophy is a putative risk factor for testicular cancer but the epidemiological evidence is not conclusive. A population based, multicenter case control study was performed from 1995 to 1997 to investigate potential risk factors for gonadal and extragonadal germ cell cancer. MATERIALS AND METHODS: The study was done in 5 German regions. Interviews were performed with 269 eligible male patients with a histologically verified diagnosis and 797 controls. Detailed information on medical and family history was collected at personal interviews. RESULTS: We identified a significantly elevated risk for testicular cancer in relation to testis and/or groin trauma (odds ratio 2.5, 95% confidence interval [CI] 1.51 to 4.20). After introducing a lag time by excluding reports of trauma within the last 12 months before diagnosis or interview the corresponding odds ratio was 2.1 (95% CI 1.24 to 3.61). Analysis of the circumstances and the reported types of injury allowed us to restrict the study to testis trauma specifically, which had an odds ratio of 3.49 (95% CI 1.78 to 6.81). To account for a potential reporting bias analysis was restricted to traumatic episodes for which medical attention was sought. This restriction resulted in an odds ratio of 0.70 (95% CI 0.19 to 2.63) after excluding from study trauma reports within the last 12 months. CONCLUSIONS: The results of our study do not support the hypothesis that testicular trauma is an important risk factor for testicular cancer. The possibility of recall bias should be considered. PMID- 11061945 TI - Immortalization of a human prostate stromal cell line using a recombinant retroviral approach. AB - PURPOSE: We established an immortalized human prostate stromal cell line with retained markers of cell differentiation and alpha1-adrenergic receptor expression. MATERIALS AND METHODS: Primary human prostate stromal explants were infected with an amphotrophic retrovirus encoding the E6/E7 open reading frame of the human papillomavirus type 16. Immunohistochemistry was used to verify the expression of prostate stromal markers. alpha1-Adrenergic receptor expression was investigated using ribonuclease protection assays and radioligand binding. Cell proliferation was measured by the WST-1 assay and cell counting. RESULTS: Clonal isolates of individual prostate stromal cells were isolated and passed in selection media. E6 and E7 expression was verified using reverse transcriptase polymerase chain reaction in the selected cell line. The new prostate stromal cell line PS30 was established which maintains the expression of alpha-smooth muscle actin and expresses 22 fmol./mg. of protein of alpha 1-adrenergic receptors, approximately equal to native human prostate alpha 1-adrenergic receptor expression. However, at a subtype level alpha 1a-adrenergic receptor expression is down-regulated and not detectable by ribonuclease protection assays or radioligand binding, while alpha 1b and alpha 1d-adrenergic receptor expression is enhanced. From a physiological prospective PS30 cells do not form tumors in nude mice and stimulation with phenylephrine does not increase cell proliferation. CONCLUSIONS: We successfully established and characterized an in vitro human prostate stromal cell line. This cell line should facilitate studies designed to characterize the role of the adrenergic nervous system in the regulation of prostate growth. PMID- 11061946 TI - Serum keratinocyte growth factor measurement in patients with prostate cancer. AB - PURPOSE: Keratinocyte growth factor (KGF) is a stromally derived growth factor important in mediating androgen induced activities in benign prostatic hyperplasia (BPH) and prostate cancer. We assessed whether serum KGF could be used as a molecular marker in patients with prostate cancer. MATERIALS AND METHODS: Using a modified double sandwich enzyme-linked immunosorbent assay, we measured serum KGF in 56 men with prostate cancer and 81 men with BPH. Comparative analyses were made with total serum prostate specific antigen (PSA), disease stage and clinical grade. RESULTS: Following optimization, a sensitive and reproducible assay for serum KGF measurement was developed. Serum KGF levels tend to be higher in men with BPH compared to those with prostate cancer (1,242 and 828 pg./ml., respectively). A weak but significant linear relationship between PSA and KGF (p = 0.034) was found in patients with BPH. There was no association between KGF and tumor grade or stage but there was a strong positive linear relationship between PSA and KGF (p = 0.006, R(2) = 68.3%) in low grade tumors. In those men with serum PSA less than 10 ng./ml. KGF levels were significantly higher in BPH compared to prostate cancer cases (965 +/- 245 and 133 +/- 61.3 pg./ml., respectively, p = 0.0058). Using a KGF threshold range of 500 to 900 pg./ml., specificity for detecting BPH was 88% to 100% and the positive predictive value was 92% to 100%. CONCLUSIONS: We have optimized a reproducible and sensitive enzyme-linked immunosorbent assay system for the measurement of serum KGF. Overall KGF levels tend to be lower in patients with prostate cancer than with BPH. In patients with serum PSA less than 10 ng./ml. serum KGF levels were significantly higher in the BPH compared to the prostate cancer group. A large prospective study is indicated to assess the role of serum KGF measurement in patients with prostate cancer. PMID- 11061947 TI - Prognostic value of cell cycle proteins p27(kip1) and MIB-1, and the cell adhesion protein CD44s in surgically treated patients with prostate cancer. AB - PURPOSE: Molecular tissue markers may give the clinician additional information about patients with prostate cancer at risk for treatment failure after retropubic radical prostatectomy. We substantiate the prognostic value of 3 tissue markers, the cell cycle proteins p27(kip1) and MIB-1, and the cell adhesion protein CD44s, in addition to more conventional pathological prognosticators in an historical (before prostate specific antigen) cohort of patients with prostate cancer. MATERIALS AND METHODS: Representative tumor sections from 92 patients who underwent retropubic radical prostatectomy were immunohistochemically stained with antibodies against p27(kip1), MIB-1 (Ki-67) and CD44s, and assessed in a semiquantitative manner. Gleason score and pathological tumor stage were recorded. All variables were correlated with clinical progression and disease specific survival on univariate and multivariate analyses. RESULTS: On univariate analysis low (less than 50%) p27(kip1), high (10% or greater) MIB-1 and loss of CD44s expression were significantly associated with clinical outcome parameters, although MIB-1 did not reach statistical significance for disease specific survival. All 3 molecules were highly correlated with Gleason score and pathological tumor stage. Multivariate analysis showed that low p27kip1 was independent of grade and stage in predicting clinical recurrence (p <0.001) and disease specific survival (p = 0.045), while loss of CD44s was an additional independent prognostic factor for clinical recurrence (p = 0.02). CONCLUSIONS: Reduced p27(kip1) expression is an independent predictor of poor outcome in prostate cancer, while MIB-1 is not. Decreased expression of CD44s yields additional information in predicting clinical recurrence. These tissue markers may identify patients at risk for disease recurrence after retropubic radical prostatectomy who may benefit from adjuvant therapy. PMID- 11061948 TI - Essential role for G proteins in prostate cancer cell growth and signaling. AB - PURPOSE: G proteins are involved in the regulation of multiple cellular functions, including metabolism and proliferation. We studied the role of Gi/o protein subunits in the growth and survival of prostate cancer cells. MATERIALS AND METHODS: We investigated the effects of pertussis toxin and the G beta gamma sequestrant peptide G protein coupled receptor kinase 2 carboxy terminus on the growth and mitogenic signaling of prostate cells. RESULTS: Pertussis toxin treatment inhibited the lysophosphatidic acid and serum mediated growth of prostate cancer PC-3 cells by 70% to 80% but showed no effect on insulin-like growth factor 1 (IGF-1) or epidermal growth factor (EGF) mediated growth of these cells. Growth and survival of cells are dependent on activation of intracellular signaling cascades, including those of the mitogen activated protein kinase and Akt pathways. Treatment of the PC-3 cells with lysophosphatidic acid, EGF or serum induced an 8-fold increase in the phosphorylation levels of the mitogen activated protein kinases Erk 1 and 2, and a 3-fold increase in the phosphorylation level of Akt. Erk 1/2 and Akt phosphorylation by lysophosphatidic acid and serum was inhibited by pertussis toxin, suggesting a Gi/o subunit dependent mechanism. EGF and IGF-1 mediated increase in phosphorylation of Erk 1/2 and Akt was independent of pertussis toxin action. Expression of the G beta gamma sequestrant peptide G protein coupled receptor kinase 2 carboxy terminus inhibited the lysophosphatidic acid and serum mediated activation of Erk 1/2 and Akt but showed no effect on the IGF-1 or EGF mediated response. Finally, we showed that activation of the Erk 1/2 pathway in the prostate cancer cells by lysophosphatidic acid and serum is dependent on the EGF receptor and c-Src protein tyrosine kinases. Whereas activation of Akt by these stimuli is not dependent on protein tyrosine kinase activation, it is mediated by PI3K. CONCLUSIONS: These data indicate that lysophosphatidic acid and serum induce proliferation and mitogenic signaling of prostate cancer cells. Importantly, the serum mediated growth of these cells is dependent on Gi beta gamma subunits, suggesting an important regulatory role for G proteins in the growth of prostate cancer cells. PMID- 11061949 TI - Prostatic levels of fatty acids and the histopathology of localized prostate cancer. AB - PURPOSE: The consumption of various fatty acids has been associated with advanced stage and fatal prostate cancer. While numerous mechanisms have been postulated, to our knowledge there physiological data linking exposure and prognosis in humans are lacking. We examined prostatic levels of individual fatty acids in relation to the prevalence of histopathological characteristics associated with invasiveness and the risk of progression in 49 men undergoing radical prostatectomy for localized prostate cancer. MATERIALS AND METHODS: Fatty acids were measured using capillary gas chromatography in fresh nonmalignant prostate tissue collected at surgery. Markers of invasiveness and increased risk of progression (Gleason sum 7 or greater, perineural invasion, anatomical or surgical margin involvement, extracapsular extension, seminal vesical involvement and stage T3 tumor) were evaluated separately. Each marker was dichotomized into a yes (case) and no (control) level with patients grouped accordingly. Mean concentrations were compared using the Wilcoxon rank sum test. RESULTS: The percent of total prostatic polyunsaturated fat and polyunsaturated-to-saturated fat ratios were significantly lower in the presence of perineural invasion, seminal vesical involvement and stage T3 tumor (p = 0.02 to 0.049). alpha Linolenic acid was significantly lower when tumor extended to an anatomical or surgical margin (p = 0.008). The omega-3 and omega-3-to-omega-6 fatty acid ratios were 1.5 to 3.3-fold lower in cases than in controls, reaching borderline significance in nearly all comparisons (p = 0.052 to 0.097). Saturated and monounsaturated fatty acids were not associated with the traits examined. CONCLUSIONS: These data suggest that polyunsaturated fatty acids and perhaps essential fatty acids in particular help to regulate prostate carcinogenesis in humans. PMID- 11061950 TI - Adenovirus mediated cytosine deaminase gene transduction and 5-fluorocytosine therapy sensitizes mouse prostate cancer cells to irradiation. AB - PURPOSE: We assess the ability of adenovirus mediated expression of the Escherichia coli cytosine deaminase gene in conjunction with the prodrug 5 fluorocytosine to result in radiation sensitization in the mouse prostate cancer cell line RM-1 in vitro. MATERIALS AND METHODS: To document cytotoxicity of gene therapy, RM-1 cells were exposed to escalating doses of adenovirus mediated cytosine deaminase and a fixed dose of 5-fluorocytosine or phosphate buffered saline. Viable cells as determined by exclusion of trypan blue were counted the following day. Cytosine deaminase expressing RM-1 cells were then irradiated as single cell suspensions at various doses of radiation in a cesium source (4.4 Gy. per minute) and randomized to receive 5-fluorocytosine therapy at different times in relation to the external radiation therapy. End points were determined in a clonogenic assay by counting colonies with greater than 50 cells 7 days after replating. RESULTS: Use of adenovirus mediated cytosine deaminase plus 5 fluorocytosine demonstrated viral dose dependent killing of RM-1 cells to a maximum of 85%, while either therapy alone was nontoxic. Neither adenovirus mediated cytosine deaminase infection nor 5-fluorocytosine alone influenced external radiation therapy killing. However, after controlling for death due to gene therapy alone, the combination of adenovirus mediated cytosine deaminase plus 5-fluorocytosine and external radiation therapy resulted in synergistic activity to approximately 2 logs of cell kill at low doses of radiation (p = 0.001). While altering the chronology of prodrug exposure in relation to external radiation therapy maintained synergy in all scenarios tested, starting 5 fluorocytosine 24 hours before external radiation therapy resulted in the most profound killing (p = 0.04), which indicates the importance of maintaining prodrug therapy during external radiation therapy. CONCLUSIONS: The combination of adenovirus mediated cytosine deaminase plus 5-fluorocytosine and radiation therapy resulted in radiation sensitization with clinically relevant doses of radiation suggesting a potential usefulness of this treatment in patients with prostate cancer. PMID- 11061951 TI - Increased prevalence and analysis of risk factors for indinavir nephrolithiasis. AB - PURPOSE: Indinavir is a protease inhibitor used for treating HIV-1. The drug is lithogenic and was thought to cause a 3% incidence of kidney stones. We evaluated a cohort of patients positive for HIV on indinavir to determine the incidence of indinavir nephrolithiasis and identify risk factors for indinavir stone formation. MATERIALS AND METHODS: Our cohort study of the prevalence of indinavir nephrolithiasis included 155 patients with HIV for 5,732 patient-weeks. The same cohort was then used for a retrospective chart review to assess patient age, weight, duration of drug use, time to stone formation, CD4 count, creatinine, alanine transaminase, and urinary pH and specific gravity as risk factors for stone formation. RESULTS: We estimated the cumulative incidence of indinavir stone formation by the Kaplan-Meier product limit estimator method. At 78 weeks 43.2% of patients had stones (95% confidence interval [CI] 0.292 to 0.543). Increasing age was the only variable that was a statistically significant predictor of indinavair urolithiasis (relative risk 0.955, 95% CI 0.918 to 0.993, p = 0.0159). The mean duration plus or minus standard deviation of indinavir use was statistically the same in each group (42.5 +/- 27. 2 and 40.3 +/- 27.1 weeks in those without and with stones, respectively) despite the observed mean time to stone formation of 23.0 +/- 19.8 weeks. CONCLUSIONS: The clinical prevalence of indinavir nephrolithiasis is much greater than initially reported. Nephrolithiasis during indinavir use does not appear to induce patients to withdraw from the drug. PMID- 11061952 TI - Hendra virus: a highly lethal zoonotic agent. PMID- 11061953 TI - Parasitic gastritis: responsibility for pathological change. PMID- 11061954 TI - Hendra (equine morbillivirus) AB - Hendra has been recognized in Australia as a new zoonotic disease of horses since 1994/5 and subsequent work has shown that the viral agent is endemic in certain species of fruit bat. The Hendra virus is the type species of a new genus within the sub-family Paramyxovirinae, which also contains another newly identified zoonotic bat virus, namely Nipah. It is assumed that contact with bats has led to the Hendra virus being transferred to horses on each of the three separate incidents that have been reported in the last five years. No evidence has been found for widespread subclinical infection of horses. Infected horses can develop a severe and often fatal respiratory disease characterized by dyspnoea, vascular endothelial damage and pulmonary oedema. Nervous signs may also occur. Fatal respiratory disease has been seen in cats and guinea pigs following experimentally induced infections. Transmission of the virus from horses to other horses or man seems to have taken place, but very close contact was required. Three human cases have been recognized, all in association with equine cases. There have been two human fatalities, one due to respiratory failure and the other from a delayed-onset encephalitis. A number of diagnostic methods have been developed, but great care must be taken in obtaining samples from suspected cases. PMID- 11061955 TI - Pathophysiology of abomasal parasitism: is the host or parasite responsible? AB - Nematode larvae developing within the glands cause local loss of parietal cells and mucous cell hyperplasia whereas reduced acid secretion, increased serum gastrin and pepsinogen concentrations and generalized histological changes are associated with parasites in the abomasal lumen. Parietal cells with dilated canaliculi and/or degenerative changes typical of necrosis are present soon after the transplantation of adult worms, and abomasal secretion is also affected. Anaerobic bacteria survive in greater numbers as the pH rises, with bacterial densities becoming similar to ruminal populations at an abomasal pH of 4 and above. Failure to lyse bacteria may affect adversely the nutrition of the host. The parasites may initiate the pathophysiology through the release of excretory/secretory (ES) products which either act directly on parietal cells or indirectly through enterochromaffin-like (ECL) cells by provoking inflammation or by disrupting the protective mucosal defence system. Parietal cell dysfunction is proposed as a key event which leads to loss of mature chief cells and mucous cell hyperplasia, as well as hypergastrinaemia. Inflammation increases circulating pepsinogen concentrations and may also contribute to increased gastrin secretion. Stimulation of mucosal proliferation and differentiation of parietal cells in the isthmus by the raised serum gastrin levels will be beneficial by generating a new population of active parietal cells and adequate acid secretion. PMID- 11061956 TI - Swine vesicular disease: an overview. AB - Swine vesicular disease (SVD) is a notifiable viral disease of pigs included on the Office International des Epizooties List A. The first outbreak of the disease was recognized in Italy in 1966. Subsequently, the disease has been reported in many European and Asian countries. The causative agent of the disease is SVD virus which is currently classified as a porcine variant of human coxsackievirus B5 and a member of the genus enterovirus in the family picornaviridae. From a clinical point of view, SVD is relatively unimportant, rarely causing deaths and usually only a minor setback to finishing schedules. However, the clinical signs which it produces are indistinguishable from those caused by foot-and-mouth disease, and its presence prevents international trade in pigs and pig products. This article reviews recent findings on all aspects of the virus and the disease which it causes. PMID- 11061957 TI - Tuberculosis in deer: perceptions, problems and progress. AB - Since the emergence of deer farming as an alternative farming enterprise over the past 30 years, there has been an increasing awareness of the potential threat posed by tuberculosis (TB) to domesticated deer. TB, caused by Mycobacterium bovis, has been found in deer in every country involved with deer farming. Different types of TB control policies, which vary from whole-herd depopulation to selective testing and slaughter of reactor animals, have been implemented. Extensive research has been carried out, incorporating modern microbiological and immunological concepts and advanced molecular methodologies, to find new solutions for the eradication of TB from domesticated deer. This work has resulted in valuable new insights into the aetiology, transmission, pathogenesis, diagnosis, prevention and heritability of resistance to M. bovis infection in ruminants. This knowledge has complemented the existing literature database on bovine and human TB and will provide new strategies for improved diagnosis, vaccination and selective breeding to control TB, which should be relevant for human, domestic livestock and wildlife populations. PMID- 11061958 TI - Studies on the experimental induction of ptosis in horses. AB - The precise appearance of ptosis due to lesions at different sites was investigated in experimental ponies. The angles of the eyelashes to the head was used as an objective measurement of ptosis after local anaesthesia of the sympathetic trunk or the palpebral nerve and the administration of an ocular alpha agonist or antagonist. It was shown that ptosis is not an inevitable consequence of palpebral nerve pathology, that ocular alpha antagonists can induce ptosis, and that alpha agonist eyedrops have an inconsistent effect on the equine pupil, but are consistent at reversing ptosis induced by sympathetic denervation in unsedated horses. PMID- 11061959 TI - Body centre of mass movement in the sound horse. AB - The body centre of mass (BCM) is a key factor in the analysis of equine locomotion, as its position and movement determines the distribution and magnitude of loads on the limbs. In this study, the three-dimensional (3D) movement of the BCM in walking and trotting horses was assessed using a kinematic, segmental method. Thirty markers representing 20 body segments were recorded in 12 sound horses while standing, walking and trotting on a treadmill using a high-speed video system. Based on segmental inertial data, 3D positions of the segmental centres of mass as well as the total BCM were calculated. The position within the trunk during square standing and the movements of the BCM were determined for the three planes. The position of the BCM in the standing horse is presented relative to external reference points. At the trot, vertical displacement amplitude of the BCM amounted to 53 (6) mm as mean (sd), which was 27% smaller than external trunk movement. Medio-lateral displacement amplitude of the BCM was 19 (4) mm, 34% less than trunk amplitude. Sagittal forward-backward oscillations of the BCM independent from general forward movement were 13 (3) mm, being 24% less than trunk movements. At the walk, vertical, medio-lateral and sagittal BCM movements were smaller than trunk movements by 43, 65 and 65% respectively. The results show reduced and efficient BCM movements compared to the trunk and form a basis for the assessment of various clinical conditions such as lameness, the influence of a rider and various dressage performances. PMID- 11061960 TI - Influence of left atrial enlargement and body weight on the development of atrial fibrillation: retrospective study on 205 dogs. AB - We studied 205 dogs with cardiac diseases associated with left atrial enlargement (LAE). On the basis of electrocardiogram results, they were divided into: Group A, 50 dogs with atrial fibrillation (AF) and Group B, 155 dogs without AF. Group B was further subdivided in Group BI (123 dogs with sinus rhythm) and Group BII (32 dogs with cardiac arrhythmias other than AF). Bodyweight (BW) and left atrial diameter (LA) of Group A dogs were significantly greater (P< 0.05) than dogs in all other groups. The left atrium/aorta (LA/Ao) ratio of Group A dogs was significantly higher (P< 0.05) than that of dogs of Group B and BI. Using the couple of variables BW and LA, the logistic regression models were able to predict "non-development of AF" (with 92.3% probability) vs. "development of AF" (with 74% probability), and "maintaining sinus rhythm" (with 95.1% probability) vs."development of AF" (with 86% probability). PMID- 11061961 TI - Use of whole blood for spectrophotometric determination of cholinesterase activity in dogs. AB - Whole blood has been compared with erythrocytes and plasma for spectrophotometric cholinesterase determination in the dog. Cholinesterase activity was characterized using two substrates: acetylthiocholine and butyrylthiocholine. Acetylcholinesterase was the only form of cholinesterase present on erythrocytes and hydrolysed only acetylthiocholine. Butyrylcholinesterase (pseudocholinesterase) was predominant in plasma, hydrolysing mainly butyrylthiocholine. Based on these results, a method based on the use of two substrates (acetylthiocholine for monitoring acetylcholinesterase and butyrylthiocholine for determining butyrylcholinesterase) in the same whole blood sample is recommended for canine cholinesterase analysis. This way of monitoring both enzymes can be easily automated, yielding good within (CVs < 5%) and between run (CVs < 7%) precision. PMID- 11061962 TI - Differences in experimental virulence of bovine viral diarrhoea viral strains isolated from haemorrhagic syndromes. AB - In the late 1980s, a new hypervirulent and epidemic form of bovine viral diarrhoea virus (BVDV) infection appeared in North America. A similar but sporadic syndrome was later reported in Europe. To compare the pathogenic characters of the North American and European hypervirulent strains, we inoculated BVDV naive calves with BVDV strains isolated from haemorrhagic syndromes originating in Belgium, France and the USA. The experimental procedure comprised daily clinical examination and measurement of blood and virological parameters. The American BVD890/256 strain induced severe thrombocytopaenia, profuse diarrhoea and pneumonia in all calves, indicating that hypervirulent BVDV could be the primary infectious agent of pneumonia. Interestingly, a strong correlation was observed between the intense viraemia and a decreased platelet count. None of the European strains tested induced significant pathological signs, although isolated from cases presenting haemorrhagic syndrome. PMID- 11061963 TI - The effect of pig farming on copper and zinc accumulation in cattle in Galicia (north-western Spain). AB - Copper and zinc are frequently added at high concentrations to pig diets as growth promoters. Livestock grazing pasture contaminated with pig slurry may, therefore, be at risk from excessive intake of these elements. High liver copper concentrations have been detected in cattle from the agricultural region of Galicia (NW Spain), especially where there is intensive pig farming. The aim of this study was to evaluate whether pig farming does affect accumulation of copper and zinc in cattle in Galicia. Hepatic copper and zinc concentrations in calves were elevated in areas with naturally high levels of these elements in the soil. The densities of young pigs (piglets and growing-finishing pigs), but not reproductive sows, also influenced copper accumulation in calves. Liver copper levels in calves were significantly and positively related to the density of young pigs in the region. In areas with the highest pig densities, more than 20% of the cattle analysed had hepatic copper concentrations that exceeded the potentially toxic concentration of 150 mg/kg fresh weight. There was no evidence that zinc accumulation in calves was affected by pig density. PMID- 11061964 TI - Effect of phenobarbitone treatment against signal grass (Brachiaria decumbens) toxicity in sheep. AB - The effect of phenobarbitone against signal grass (Brachiaria decumbens) toxicity was studied in 26 male crossbred sheep. Grazing on signal grass significantly decreased the concentration of cytochrome P-450 and the activity of drug metabolizing enzymes, viz. aminopyrine-N-demethylase, aniline-4-hydroxylase, UDP- glucuronyltransferase and glutathione-S-transferase in liver and kidneys of affected sheep.Oral administration of phenobarbitone (30 mg/kg body weight) for five consecutive days before grazing on B. decumbens pasture, and thereafter, for three consecutive days every two weeks, resulted in significant increases in hepatic and renal activities of drug-metabolizing enzymes. The induction of drug metabolizing activity in sheep grazing on signal grass group was found to be lower than in animals given phenobarbitone alone. Induction by phenobarbitone provided a degree of protection against the toxic effects of B. decumbens as indicated by the delay in the appearance of signs of toxicity. Furthermore, these were much milder compared to those in the sheep not treated with phenobarbitone. The present study suggests that phenobarbitone-type cytochrome P-450 isoenzyme induction may increase resistance against signal grass (B. decumbens) toxicity in sheep. PMID- 11061965 TI - Different proteasome subtypes in a single tissue exhibit different enzymatic properties. AB - It is concluded from many experiments that mammalian tissues and cells must contain a heterogeneous population of 20 S proteasome complexes. We describe the purification and separation by chromatographic procedures of constitutive 20 S proteasomes, 20 S immuno-proteasomes and intermediate-type 20 S proteasomes from a given tissue. Our data demonstrate that each of these three groups comprises more than one subtype and that the relative ratios of the subtypes differ between different rat tissues. Thus, six subtypes could be identified in rat muscle tissue. Subtypes I and II are constitutive proteasomes, while subtypes V and VI comprise immuno-proteasomes. Subtypes III and IV belong to a group of intermediate-type proteasomes. The subtypes differ with regard to their enzymatic characteristics. Subtypes I-III exhibit high chymotrypsin-like activity and high peptidylglutamylpeptide hydrolysing activity, while these activities are depressed in subtypes IV-VI. In contrast, trypsin-like activity of subtypes IV-VI is enhanced in comparison to subtypes I-III. Importantly, the subtypes also differ in their preferential cleavage site usage when tested by digestion of a synthetic 25mer polypeptide substrate. Therefore, the characteristics of proteasomes purified from tissues or cells represent the average of the different subtype activities which in turn may have different functions in vivo. PMID- 11061966 TI - Heptameric ring structure of the heat-shock protein ClpB, a protein-activated ATPase in Escherichia coli. AB - The heat-shock protein ClpB is a protein-activated ATPase that is essential for survival of Escherichia coli at high temperatures. ClpB has also recently been suggested to function as a chaperone in reactivation of aggregated proteins. In addition, the clpB gene has been shown to contain two translational initiation sites and therefore encode two polypeptides of different size. To determine the structural organization of ClpB, the ClpB proteins were subjected to chemical cross-linking analysis and electron microscopy. The average images of the ClpB proteins with end-on orientation revealed a seven-membered, ring-shaped structure with a central cavity. Their side-on view showed a two-layered structure with an equal distribution of mass across the equatorial plane of the complex. Since the ClpB subunit has two large regions containing consensus sequences for nucleotide binding, each layer of the ClpB heptamer appears to represent the side projection of one of the major domains arranged on a ring. In the absence of salt and ATP, the ClpB proteins showed a high tendency to form a heptamer. However, they dissociated into various species of oligomers with smaller sizes, depending on salt concentration. Above 0.2 M NaCl, the ClpB proteins behaved most likely as a monomer in the absence of ATP, but assembled into a heptamer in its presence. Furthermore, mutations of the first ATP-binding site, but not the second site, prevented the ATP-dependent oligomerization of the ClpB proteins in the presence of 0.3 M NaCl. These results indicate that ClpB has a heptameric ring-shaped structure with a central cavity and this structural organization requires ATP binding to the first nucleotide-binding site localized to the N-terminal half of the ATPase. PMID- 11061967 TI - Molecular-level thermodynamic and kinetic parameters for the self-assembly of apoferritin molecules into crystals. AB - The self-assembly of apoferritin molecules into crystals is a suitable model for protein crystallization and aggregation; these processes underlie several biological and biomedical phenomena, as well as for protein and virus self assembly. We use the atomic force microscope in situ, during the crystallization of apoferritin to visualize and quantify at the molecular level the processes responsible for crystal growth. To evaluate the governing thermodynamic parameters, we image the configuration of the incorporation sites, "kinks", on the surface of a growing crystal. We show that the kinks are due to thermal fluctuations of the molecules at the crystal-solution interface. This allows evaluation of the free energy of the intermolecular bond phi=3.0 k(B)T=7.3 kJ/mol. The crystallization free energy, extracted from the protein solubility, is -42 kJ/mol. Published determinations of the second virial coefficient and the protein solubility between 0 and 40 degrees C revealed that the enthalpy of crystallization is close to zero. Analyses based on these three values suggest that the main component in the crystallization driving force is the entropy gain of the water molecules bound to the protein molecules in solution and released upon crystallization. Furthermore, monitoring the incorporation of individual molecules in to the kinks, we determine the characteristic frequency of attachment of individual molecules at one set of conditions. This allows a correlation between the mesoscopic kinetic coefficient for growth and the molecular-level thermodynamic and kinetic parameters determined here. We found that step growth velocity, scaled by the molecular size, equals the product of the kink density and attachment frequency, i.e. the latter pair are the molecular level parameters for self-assembly of the molecules into crystals. PMID- 11061968 TI - The interaction of the B-ring of colchicine with alpha-tubulin: a novel footprinting approach. AB - Tubulin, the major structural component of the microtubules, participates actively in mitotic spindle formation and chromosomal organization during cell division. Tubulin is the major target for a variety of anti-mitotic drugs. Some of the drugs, such as Vinca alkaloids and taxol, are routinely used for cancer chemotherapy. It is unfortunate that our knowledge of the binding sites on tubulin of these drugs is limited because of lack of a useful and appropriate tool. The photoaffinity labeling approach is the major technique available at present to detect the binding sites of drugs on tubulin. This method, however, has several limitations. First, only part of the binding site can be identified, namely, the residues which react with the photoaffinity label. Second, there are regions of tubulin which are not at the binding site but are affected by the binding of the drug; these regions can not be detected by the photoaffinity labeling approach. The third, and perhaps most serious, limitation is that the traditional approach can detect areas which have nothing to do with the binding of the ligand but which are within a certain distance of the binding site, that distance being less than the length of the photoreactive moiety attached to the ligand. There has been a great deal of controversy on the localization of the binding site of colchicine on tubulin, with some reports suggesting that the binding site is on alpha and some supporting a binding site on beta. Colchicine also has significant effects on tubulin conformation, but the regions which are affected have not been identified. We have attempted here to address these questions by a novel "footprinting" method by which the drug-binding sites and as well as the domain of tubulin affected by drug-induced conformational changes could be determined. Here, we report for the first time that the interaction of the B-ring of colchicine with the alpha-subunit affects a domain of tubulin which appears to be far from its binding site. This domain includes the cysteine residues at positions 295, 305, 315 and 316 on alpha-tubulin; these residues are located well away from the alpha/beta interface where colchicine appears to bind. This is correlated with the stabilizing effect of colchicine on the tubulin molecule. Furthermore, we also found that the B-ring of colchicine plays a major role in the stability of tubulin while the A and the C-rings have little effect on it. Our results therefore, support a model whereby colchicine binds at the alpha/beta interface of tubulin with the B-ring on the alpha-subunit and the A and the C-rings on the beta-subunit. PMID- 11061969 TI - How the multifunctional yeast Rap1p discriminates between DNA target sites: a crystallographic analysis. AB - Rap1p from Saccharomyces cerevisiae is a multifunctional, sequence-specific, DNA binding protein involved in diverse cellular processes such as transcriptional activation and silencing, and is an essential factor for telomere length regulation and maintenance. In order to understand how Rap1p discriminates between its different DNA-binding sites, we have determined the crystal structure of the DNA-binding domain of the Rap1p (Rap1pDBD) in complex with two different DNA-binding sites. The first DNA sequence is the HMRE binding site found at silencers, which contains four base-pair substitutions in comparison to the telomeric binding site present in our earlier crystal structure of the Rap1pDBD TeloA complex. The second complex contains an alternative telomeric binding site, TeloS, in which two half-sites are spaced closer together than in the TeloA complex. The determination of these structures was complicated by the presence of merohedral twinning in the crystals. Through identification of the twinning operator and determination of the twin fraction of the crystals, we were able to deconvolute the twinned intensities into their untwinned components, and to calculate electron density maps for both complexes. The structural information shows that the two domains present in the Rap1pDBD bind to these two biologically relevant binding sites through subtle side-chain movements at the protein-DNA interface, rather than through global domain rearrangements. PMID- 11061970 TI - The homologous pairing domain of RecA also mediates the allosteric regulation of DNA binding and ATP hydrolysis: a remarkable concentration of functional residues. AB - Switching between the active (ATP and DNA bound) and inactive conformations of the homologous recombination RecA protein is regulated by ATP hydrolysis. First, we use the homologous pairing domain of RecA derived from its mobile loop L2 to show that the interaction of this random coil peptide with the gamma-phosphate of ATP results in a peptide beta-conformation similar to that previously shown to be induced by DNA binding. Next, we show that in the whole RecA protein two residues in this L2 domain, Gln194 and Arg196, are catalytic amino acid residues for ATP hydrolysis and functionally resemble the corresponding residues engaged in GTP hydrolysis by two distinct classes of G proteins. Finally, we show that the role of DNA and high salt in the stimulation of the ATPase of RecA is to stabilize this highly mobile region involved in hydrolysis. This is a role similar to that described for RGSs in the activation of the GTPase of heterotrimeric G proteins. Therefore, (i) a prototypical DNA-dependent ATPase and ATP-stimulated DNA-binding protein, RecA, and eukaryotic signaling proteins share common stereochemical regulatory mechanisms; and (ii) in a remarkable example of parsimony, loop L2 is a molecular switch that controls both ATP promoted DNA binding and pairing reactions and DNA stimulated ATP hydrolysis. PMID- 11061971 TI - Crystallographic analysis reveals common modes of binding of medium and long chain fatty acids to human serum albumin. AB - Human serum albumin (HSA) is an abundant plasma protein that is responsible for the transport of fatty acids. HSA also binds and perturbs the pharmacokinetics of a wide range of drug compounds. Binding studies have revealed significant interactions between fatty acid and drug-binding sites on albumin but high resolution structural information on ligand binding to the protein has been lacking. We report here a crystallographic study of five HSA-fatty acid complexes formed using saturated medium-chain and long-chain fatty acids (C10:0, C12:0, C14:0, C16:0 and C18:0). A total of seven binding sites that are occupied by all medium-chain and long-chain fatty acids have been identified, although medium chain fatty acids are found to bind at additional sites on the protein, yielding a total of 11 distinct binding locations. Comparison of the different complexes reveals key similarities and significant differences in the modes of binding, and serves to rationalise much of the biochemical data on fatty acid interactions with albumin. The two principal drug-binding sites, in sub-domains IIA and IIIA, are observed to be occupied by fatty acids and one of them (in IIIA) appears to coincide with a high-affinity long-chain fatty acid binding site. PMID- 11061972 TI - Crystal structure of N-acyl-D-glucosamine 2-epimerase from porcine kidney at 2.0 A resolution. AB - The X-ray crystallographic structure of N-acyl-d-glucosamine 2-epimerase (AGE) from porcine kidney, which has been identified to be a renin-binding protein (RnBP), was determined by the multiple isomorphous replacement method and refined at 2.0 A resolution with a final R-factor of 16.9 % for 15 to 2.0 A resolution data. The refined structure of AGE comprised 804 amino acid residues (one dimer) and 145 water molecules. The dimer of AGE had an asymmetric unit with approximate dimensions 46 Ax48 Ax96 A. The AGE monomer is composed of an alpha(6)/alpha(6) barrel, the structure of which is found in glucoamylase and cellulase. One side of the AGE alpha(6)/alpha(6)-barrel structure comprises long loops containing five short beta-sheets, and contributes to the formation of a deep cleft shaped like a funnel. The putative active-site pocket and a possible binding site for the substrate N-acetyl-d-glucosamine (GlcNAc) were found in the cleft. The other side of the alpha(6)/alpha(6)-barrel comprises short loops and contributes to the dimer formation. At the dimer interface, which is composed of the short loops and alpha-helices of the subunits, five strong ion-pair interactions were observed, which play a major role in the dimer assembly. This completely ruled out the previously accepted hypothesis that the formation of the RnBP homodimer and RnBP renin heterodimer requires the leucine zipper motif present in RnBP. PMID- 11061973 TI - X-ray structures of five renin inhibitors bound to saccharopepsin: exploration of active-site specificity. AB - Saccharopepsin is a vacuolar aspartic proteinase involved in activation of a number of hydrolases. The enzyme has great structural homology to mammalian aspartic proteinases including human renin and we have used it as a model system to study the binding of renin inhibitors by X-ray crystallography. Five medium-to high resolution structures of saccharopepsin complexed with transition-state analogue renin inhibitors were determined. The structure of a cyclic peptide inhibitor (PD-129,541) complexed with the proteinase was solved to 2.5 A resolution. This inhibitor has low affinity for human renin yet binds very tightly to the yeast proteinase (K(i)=4 nM). The high affinity of this inhibitor can be attributed to its bulky cyclic moiety spanning P(2)-P(3)' and other residues that appear to optimally fit the binding sub-sites of the enzyme. Superposition of the saccharopepsin structure on that of renin showed that a movement of the loop 286-301 relative to renin facilitates tighter binding of this inhibitor to saccharopepsin. Our 2.8 A resolution structure of the complex with CP-108,420 shows that its benzimidazole P(3 )replacement retains one of the standard hydrogen bonds that normally involve the inhibitor's main-chain. This suggests a non-peptide lead in overcoming the problem of susceptible peptide bonds in the design of aspartic proteinase inhibitors. CP-72,647 which possesses a basic histidine residue at P(2), has a high affinity for renin (K(i)=5 nM) but proves to be a poor inhibitor for saccharopepsin (K(i)=3.7 microM). This may stem from the fact that the histidine residue would not bind favourably with the predominantly hydrophobic S(2) sub-site of saccharopepsin. PMID- 11061974 TI - A snapshot of a transition state analogue of a novel thermophilic esterase belonging to the subfamily of mammalian hormone-sensitive lipase. AB - EST2 is a novel thermophilic carboxylesterase, isolated and cloned from Alicyclobacillus (formerly Bacillus) acidocaldarius, which optimally hydrolyses esters with acyl chain lengths of six to eight carbon atoms at 70 degrees C. On the basis of the amino acid sequence homology, it has been classified as a member of the mammalian hormone-sensitive lipase (HSL) subfamily. The crystal structure of EST2, complexed with a sulphonyl derivative, has been determined at 2.6 A resolution by a multiple wavelength anomalous diffraction experiment on a seleno methionine derivative. EST2 presents a canonical alpha/beta hydrolase core, shielded at the C-terminal side by a cap region built up of five helices. It contains the lipase-like catalytic triad, Ser155, His282 and Asp252, whereby the nucleophile is covalently modified. This allows an unambiguous view of the putative active site of EST2, detecting the oxyanion hole, in whose formation the amino acid sequence motif His81-Gly82-Gly83-Gly84 is involved, and the hydrophobic binding pocket for the acyl chain. The structural model here reported provides the first example of a transition state analogue of an esterase/lipase belonging to the HSL group, thus affording useful information for the design of medical inhibitors. Moreover, as the first X-ray structure of a thermophilic carboxylesterase, the comparison with its mesophilic homologue, the Brefeldin A esterase (BFAE) from Bacillus subtilis, allows the identification of putative determinants of thermal stability. PMID- 11061975 TI - Multiple-step kinetic mechanism of DNA-independent ATP binding and hydrolysis by Escherichia coli replicative helicase DnaB protein: quantitative analysis using the rapid quench-flow method. AB - The kinetic mechanism of DNA-independent binding and hydrolysis of ATP by the E. coli replicative helicase DnaB protein has been quantitatively examined using the rapid quench-flow technique. Single-turnover studies of ATP hydrolysis, in a non interacting active site of the helicase, indicate that bimolecular association of ATP with the site is followed by the reversible hydrolysis of nucleotide triphosphate and subsequent conformational transition of the enzyme-product complex. The simplest mechanism, which describes the data, is a three-step sequential process defined by:?eqalign???rm Helicase+ATP?&?mathop??rightleftharpoons? ?k_1?_?k_?-1????rm (H ATP)??mathop??rightleftharpoons? ?k_2?_?k_?-2????rm (H-ADP?cdot Pi)??cr &?mathop??rightleftharpoons? ?k_3?_?k_?-3????rm (H-ADP?cdot Pi)? *?The sequential character of the mechanism excludes conformational transitions of the DnaB helicase prior to ATP binding. Analysis of relaxation times and amplitudes of the reaction allowed us to estimate all rate and equilibrium constants of partial steps of the proposed mechanism. The intrinsic binding constant for the formation of the (H-ATP) complex is K(ATP)=(1.3+/-0.5)x10(5) M(-1). The analysis of the data indicates that a part of the ATP binding energy originates from induced structural changes of the DnaB protein-ATP complex prior to ATP hydrolysis. The equilibrium constant of the chemical interconversion is K(H)=k(2)/k(-2) approximately 2 while the subsequent conformational transition is characterized by K(3)=k(3)/k(-3) approximately 30. The low value of K(H) and the presence of the subsequent energetically favorable conformational step(s) strongly suggest that free energy is released from the enzyme-product complex in the conformational transitions following the chemical step and before the product release.The combined application of single and multiple-turnover approaches show that all six nucleotide-binding sites of the DnaB hexamer are active ATPase sites. Binding of ATP to the DnaB hexamer is characterized by the negative cooperativity parameter sigma=0.25(+/-0.1). The negative cooperative interactions predominantly affect the ground state of the enzyme-ATP complex. The significance of these results for the mechanism of the free energy transduction of the DnaB helicase is discussed. PMID- 11061976 TI - How do substrates enter and products exit the buried active site of cytochrome P450cam? 1. Random expulsion molecular dynamics investigation of ligand access channels and mechanisms. AB - Cytochrome P450s form a ubiquitous protein family with functions including the synthesis and degradation of many physiologically important compounds and the degradation of xenobiotics. Cytochrome P450cam from Pseudomonas putida has provided a paradigm for the structural understanding of cytochrome P450s. However, the mechanism by which camphor, the natural substrate of cytochrome P450cam, accesses the buried active site is a long-standing puzzle. While there is recent crystallographic and simulation evidence for opening of a substrate access channel in cytochrome P450BM-3, for cytochrome P450cam, no such conformational changes have been observed either in different crystal structures or by standard molecular dynamics simulations. Here, a novel simulation method, random expulsion molecular dynamics, is presented, in which substrate-exit channels from the buried active site are found by imposing an artificial randomly oriented force on the substrate, in addition to the standard molecular dynamics force field. The random expulsion molecular dynamics method was tested in simulations of the substrate-bound structure of cytochrome P450BM-3, and then applied to complexes of cytochrome P450cam with different substrates and with product. Three pathways were identified, one of which corresponds to a channel proposed earlier on the basis of crystallographic and site-directed mutagenesis data. Exit via the water-filled channel, which was previously suggested to be a product exit channel, was not observed. The pathways obtained by the random expulsion molecular dynamics method match well with thermal motion pathways obtained by an analysis of crystallographic B-factors. In contrast to large backbone motions (up to 4 A) observed in cytochrome P450BM-3 for the exit of palmitoleic acid, passage of camphor through cytochrome P450cam only requires small backbone motions (less than 2.4 A) in conjunction with side-chain rotations. Concomitantly, in almost all the exit trajectories, salt-links that have been proposed to act as ionic tethers between secondary structure elements of the protein, are perturbed. PMID- 11061977 TI - How do substrates enter and products exit the buried active site of cytochrome P450cam? 2. Steered molecular dynamics and adiabatic mapping of substrate pathways. AB - Three possible channels by which substrates and products can exit from the buried active site of cytochrome P450cam have been identified by means of random expulsion molecular dynamics simulations. In the investigation described here, we computed estimates of the relative probabilities of ligand passage through the three channels using steered molecular dynamics and adiabatic mapping. For comparison, the same techniques are also applied to investigate substrate egress from cytochrome P450-BM3. The channel in cytochrome P450cam, for which there is the most supporting evidence from experiments (which we name pathway 2a), is computed to be the most probable ligand exit channel. It has the smallest computed unbinding work and force. For this channel, the ligand exits between the F/G loop and the B' helix. Two mechanistically distinct, but energetically similar routes through this channel were observed, showing that multiple pathways along one channel are possible. The probability of ligand exit via the next most probable channel (pathway 3), which is located between the I helix and the F and G helices, is estimated to be less than 1/10 of the probability of exit along pathway 2a. Low-frequency modes of the protein extracted from an essential dynamics analysis of a 1 ns duration molecular dynamics simulation of cytochrome P450cam with camphor bound, support the opening of pathway 2a on a longer timescale. On longer timescales, it is therefore expected that this pathway becomes more dominant than estimated from the present computations. PMID- 11061978 TI - Formation of fibrillar multimers of oat beta-glucosidase isoenzymes is mediated by the As-Glu1 monomer. AB - Oat beta-glucosidase (EC 3.2.1.21) exists in two isomeric forms of homomultimer (type I) and heteromultimer (type II), which are comprised of two 60 kDa monomers of As-Glu1 and As-Glu2. The cDNA of As-Glu2 was cloned in this study, whereas As Glu1 was previously cloned as As-P60. The As-Glu2 cDNA encodes a plastid directing transit peptide of 57 amino acid residues and a mature protein of 521 amino acid residues. The amino acid sequence of As-Glu2 is highly homologous to that of As-Glu1, except for their C-terminal portions. When the two cDNAs of the mature proteins were expressed as T7.Tag-fused proteins in Escherichia coli, they produced soluble and enzymatically active T7.Tag-As-Glu1 and T7.Tag-As-Glu2 proteins. The T7.Tag-As-Glu1 was assembled into a donut-shaped hexamer ring which was in turn stacked in integer numbers to form long fibrillar homomultimers of different lengths with a molecular mass of up to several million daltons. On the other hand, the T7.Tag-As-Glu2 primarily formed a dimer rather than a multimer. When both cDNAs of As-Glu1 and As-Glu2 were co-expressed as T7.Tag-fused mature proteins, they were also assembled into a hexamer ring comprised of the two monomers in a 1:1 stoichiometry. The heteromeric hexamer was stacked in smaller numbers to form the heteromultimer of T7. Tag-As-Glu1 and -As-Glu2. The results indicate that the As-Glu1 monomer plays a crucial role in the formation of both the As-Glu1 homomultimer and the As-Glu1 and As-Glu2 heteromultimer. We describe here a unique structure for the oat beta-glucosidase fibrillar multimer that is formed by stacking the hexamer rings composed of As-Glu1 and/or As-Glu2. PMID- 11061979 TI - Parenteral antibiotic therapy in the treatment of lower respiratory tract infections. Strategies to minimize the development of antibiotic resistance. AB - Antibiotic use is often imputed for increases in the prevalence of infections due to antibiotic-resistant bacteria. Resistance depends on the variety of genotypes in the large bacterial population and also on the selective pressures that are produced along the antibiotic concentration gradients in the body. In effect, at certain selective concentrations the antibiotic eliminates the susceptible majority, leaving a selected remainder intact. Therefore, the choice of antibiotics for the treatment of lower respiratory tract infections should take into consideration not only their effectiveness but also the pharmacokinetics of each agent and its delivery schedule. In fact, the potential therapeutic efficacy of an antibiotic depends not only on its spectrum of action, but also on the concentration it reaches at the site of infection. Most infections occur in the tissues of the body rather than in the blood and that it is accepted that appropriate antibiotic therapy requires the maintenance of significant concentrations of antibiotics at the site of infection in the lung long enough to eliminate the invading pathogen. Thus, the development of dosing schedules for most antimicrobials has been based on the postulate that drug levels need to be above the minimal inhibitory concentration (MIC) at this site for most or all the dosing interval. The selection of antimicrobial resistance appears to be strongly associated with suboptimal antimicrobial exposure, defined as an AUIC(0-24)/MIC ratio of less than 100O125. Antimicrobial regimens that do not achieve these values cannot prevent the selective pressure that leads to overgrowth of resistant bacterial subpopulations. It has been suggested that resistance can be avoided with attention to dosing, since dosing which provides an AUIC(0-24)/MIC ratio of at least 100 appears to reduce the rate of the development of bacterial resistance. Unfortunately, very different serum or lung concentration profiles can result in the same AUIC(0-24)/MIC. High doses administered sufficiently may often completely prevent any possibility of attaining a selective concentration. Alternatively, an antibiotic which has good bactericidal potency and maintains tissue and/or serum concentrations greater than the MIC or, better, minimal bactericidal concentration (MBC) throughout the dosing interval is equally effective in minimizing the development of antibiotic resistance. PMID- 11061980 TI - Transforming growth factor-beta1 induces endothelin-1 in a bovine pulmonary artery endothelial cell line and rat lungs via cAMP. AB - We investigated the mechanism of Endothelin-1 regulation by transforming growth factor-beta1 (TGF-beta1) in bovine pulmonary artery endothelial cells (BPAECs) and in isolated perfused rat lungs. Our data show that TGF-beta1 induces ET-1 gene expression and ET-1 peptide synthesis in BPAECs. The induction of preproET-1 mRNA level was due to de novo transcription, as well as mRNA stabilization, and new protein synthesis was not required for this induction. To investigate the role of cAMP-protein kinase A pathway in TGF-beta1-stimulated-ET-1 induction, we exposed BPAECs to various compounds which modulate this pathway. Dibutyryl-cAMP led to an increase in preproET-1 mRNA and Rp-cAMP abolished the induction of preproET-1 mRNA and ET-1 peptide by TGF-beta1. TGF-beta1 increased cAMP in BPAECs. Dexamethasone up-regulated preproET-1 mRNA expression and ET-1 peptide synthesis under basal and TGF-beta1-stimulated conditions. In isolated perfused rat lungs, TGF-beta1 increased preproET-1 mRNA abundance whereas Rp-cAMP inhibited the TGF-beta1-induced ET-1 gene activation. Thus our data suggest that TGF-beta1 stimulates ET-1 gene expression in BPAECs and in rat lungs via a cAMP dependent mechanism. PMID- 11061981 TI - Allergic airway hyperresponsiveness and eosinophil infiltration is reduced by a selective iNOS inhibitor, 1400W, in mice. AB - Nitric oxide (NO) hyperproduction has been reported in asthmatic airways and may contribute to airway inflammatory responses. The purpose of this study was to examine the role of NO via inducible NO synthase (iNOS) in allergic airway inflammation using a selective iNOS inhibitor, N-[3-(aminomethyl)benzyl] acetamidine (1400W), in ovalbumin (OVA)-sensitized Balb/c mice. Sensitized animals were challenged with aerosolized 0.5% OVA for 1 h on two occasions 4 h apart. 1400W or the vehicle was administered by osmotic mini-pump from 2 h before to 24 h after OVA challenge. Twenty-four hours after OVA challenge, the vehicle treated mice showed a significant airway hyperresponsiveness to intravenous methacholine (P<0.05) as well as an influx of eosinophils into the airways (P<0.05). iNOS immunoreactivity was obvious in the epithelial and, to a lesser extent, the infiltrated inflammatory cells. iNOS protein in the airway assessed by Western blotting also increased. Pretreatment with 1400W almost completely abolished the OVA-induced airway hyperresponsiveness and to a lesser extent eosinophil accumulation into the airways. These results suggest that NO synthesized by iNOS may participate in airway hyperresponsiveness and eosinophil infiltration into the airways after allergic reaction. PMID- 11061982 TI - Role of the 5-HT(2A)receptor and alpha(1)-adrenoceptor in the contractile response of rat pulmonary artery to 5-HT in the presence and absence of nitric oxide. AB - This study investigated the role of 5-HT(2A)receptors and alpha(1)-adrenoceptors in the contractile response to 5-HT in the first branch pulmonary artery of the rat and their interaction with endogenous nitric oxide. 5-HT and phenylephrine induced concentration-dependent contractions. The alpha(1)-adrenoceptor antagonists prazosin, HV723 and phentolamine produced concentration-dependent rightward shifts of the 5-HT concentration-response curves (CRC) consistent with an action at alpha(1)-adrenoceptors. The 5-HT(2)receptor antagonists ritanserin, ketanserin and methysergide produced rightward shifts that were less than would have been predicted for an action solely at 5-HT(2A)receptors. 5-HT and phenylephrine CRCs were shifted to the left by l -NAME. Endothelium denudation also increased the tissue sensitivity to 5-HT. In the presence of l -NAME, ketanserin produced greater antagonism of the 5-HT CRC but not the phenylephrine CRC. Ketanserin also produced greater antagonism of the 5-HT CRC in endothelium denuded rings compared with endothelium intact rings. These findings indicate (a) that both the alpha(1)-adrenoceptor class and the 5-HT(2A)receptor is involved in the contractile response to 5-HT; (b) in the presence of endogenous nitric oxide the contractile response to 5-HT is mediated predominently by alpha(1) adrenoceptors; (c) inhibition of endogenous nitric oxide potentiates the 5 HT(2A)receptor-mediated component of the contraction. PMID- 11061983 TI - Duration of action of inhaled vs. Intravenous beta(2)-adrenoceptor agonists in an anaesthetized guinea-pig model. AB - We compared the duration of action of the short-acting alpha(2)-adrenoceptor agonist salbutamol and the long-acting alpha(2)-adrenoceptor agonists salmeterol and formoterol when administered iv or by inhalation in a histamine-induced bronchoconstriction model in the guinea-pig. Following aerosol dosing, maximal bronchoprotector effects were seen for salbutamol, salmeterol and formoterol at concentrations of 1 mg/ml, 100 microg/ml and 30 microg/ml respectively, giving a potency order of formoterol > salmeterol > salbutamol. All displayed similar maximum effects in this system. A maximal concentration of salbutamol showed bronchoprotection at 1 h but not at 3 h post-dosing whereas maximal concentrations of formoterol and salmeterol showed protection up to 5 h post aqueous-aerosol dosing, giving a duration order of salmeterol > formoterol > salbutamol. All three alpha(2)-adrenoceptor agonists showed dose-dependent bronchoprotection and duration of action following intravenous administration; salbutamol and salmeterol were equipotent and both were less potent than formoterol. Bronchoprotection obtained with sub-maximal concentrations of all three alpha(2)-adrenoceptor agonists faded within 30 min following iv administration, but this could be extended by increasing the doses. These results demonstrate that the route of administration is important in determining the duration of action of alpha(2)-adrenoceptor agonists in the lung. Furthermore, such findings lend support to the suggestion that the physico-chemical characteristics of salmeterol govern its duration of action rather than sustained binding of this agonist to a alpha(2)-adrenoceptor exo-site. PMID- 11061984 TI - CGS 21680, dibutyryl cyclic AMP and rolipram attenuate the pro-inflammatory interactions of the Pseudomonas aeruginosa -derived pigment, 1-hydroxyphenazine, with human neutrophils. AB - The effects of the intracellular adenosine 3':5' cyclic monophosphate (cAMP) elevating agents, CGS 21680 (0.01- 1 microM) and rolipram (0.01-1 microM), as well as those of dibutyryl cAMP (0. 05-4 mM) on the pro-inflammatory interactions of the P. aeruginosa -derived pigment, 1-hydroxyphenazine (1-hp, 3.1 and 12.5 microM), with human neutrophils have been investigated in vitro. Ca(2+)fluxes in FMLP-activated neutrophils were measured using a fura-2/AM spectrofluorimetric procedure, while a colourimetric method was used to measure release of the primary granule enzyme, elastase, from the cells. Treatment with 1-hp resulted in delayed clearance of Ca(2+)from the cytosol of N -formyl- L -methionyl- L -leucyl L -phenylalanine (FMLP, 1 microM)-activated neutrophils and increased release of elastase. All 3 test agents caused dose-related antagonism of 1-hp-mediated potentiation of elastase release from activated neutrophils, which was associated with restoration of Ca(2+)homeostasis. These observations demonstrate the potential of cAMP-elevating agents, acting on Ca(2+)clearance mechanisms in activated neutrophils, to attenuate the potentially harmful pro-inflammatory effects of 1-hp. PMID- 11061986 TI - Volume 13 index PMID- 11061985 TI - Lung function improvement in smokers suffering from COPD with zafirlukast, a CysLT(1)-receptor antagonist. AB - The present study was designed to evaluate the bronchodilating role of zafirlukast, a CysLT(1)receptor antagonist, at the standard dosage currently recommended in the marketing of this agent in smokers with COPD. The study was performed using a double-blind, cross-over, randomized design and was conducted on 2 non-consecutive days. Sixteen outpatients suffering from stable COPD received 40 mg oral zafirlukast, or placebo. Lung function was controlled before drug administration and 30, 60, 120, 180, 240 min thereafter. At the end of the 4 h period, each patient received 400 microg inhaled salbutamol and spirometric testing was performed 30 min later. Zafirlukast, but not placebo, produced a significant (P<0.05) bronchodilation between 30 min and 4 h following administration, with a maximum mean increase in FEV(1)of 0.134 l (11.2%) above baseline after 2 h. Nine of 16 patients showed an increase in FEV(1)of at least 15% above baseline after zafirlukast. The maximum mean increase in FEV(1)after zafirlukast in these subjects, who were considered responders, observed after 2 h, was 0.221 (19.4%). The mean difference of post-salbutamol FEV(1)values after zafirlukast and placebo (-0.036 l) was not significant (P<0.05). In responders, the mean of differences in pre- and post-salbutamol FEV(1)values after zafirlukast was 0.077 l, whereas the mean of differences between post-salbutamol values after zafirlukast and those after placebo was -0.064 l. The mean AUC(0-4 h)for all patients was 0.121 l for placebo and 0.385 l for zafirlukast. The difference between the placebo and zafirlukast AUC(0-4 h)was significant (P<0.05). The individual FEV(1)AUC(0-4 h)after zafirlukast were higher than those after placebo in 12 out of 16 patients. These findings suggest that cysteinyl leukotrienes might be one of the causes of persistent bronchoconstriction in COPD, at least in several smokers, but do not confirm the hypothesis that the effects of zafirlukast and salbutamol are independent and additive. PMID- 11061987 TI - Identification of a novel taxol-sensitive kinase activity associated with the cytoskeleton. AB - The microtubule-targeted drug, taxol, enhances assembly of alphabeta tubulin dimers into microtubules. Recent work has established that taxol also elicits diverse effects on intracellular signaling. In-gel kinase assays with myelin basic protein as substrate revealed that taxol treatment significantly (P 5-fold increase in tripalmitin mass and a 60% decrease in polyunsaturated triacylglycerol molecular species mass (i.e. triacylglycerols containing at least one acyl residue with more than two double bonds); (ii) a 46% increase in myocardial phosphatidylinositol mass; (iii) a 44% increase in myocardial plasmenylethanolamine mass and (iv) a 22% decrease in 1-stearoyl-2-arachidonoyl phosphatidylethanolamine content. Each of the changes in phospholipid classes, subclasses and individual molecular species were prevented by insulin treatment after induction of the diabetic state. In sharp contrast, the alterations in triacylglycerol molecular species were not preventable by peripheral insulin treatment after induction of the diabetic state. These results segregate diabetes induced alterations in myocardial lipid metabolism into changes that can be remedied or not by routine peripheral insulin treatment and suggest that peripheral insulin therapy alone may not be sufficient to correct all of the metabolic alterations present in diabetic myocardium. PMID- 11062061 TI - Effect of alpha-1 antitrypsin Portland variant (alpha 1-PDX) on HIV-1 replication. AB - The present work investigated the potential role of alpha-1 antitrypsin Portland variant (alpha 1-PDX), a bioengineered serine proteinase inhibitor (serpin), in the interference with the viral replication of HIV-1, induction of syncytia and maturation of envelope glycoprotein gp160 to gp120 and gp41. A Jurkat lymphoid cell line transfected with a plasmid containing the alpha 1-PDX cDNA (J-PDX) and expressing the protein in a stable manner was infected with HIV-1(Lai). Controls were Jurkat cells transfected with the same vector pcDNA3 without the cDNA insert (J-pcDNA3). The results showed that viral replication of HIV-1 was significantly inhibited with a delay in replication kinetics in J-PDX cells as compared with J pcDNA3 cells. In addition, a comparison of the infectious capacity of viruses produced in the presence and absence of alpha 1-PDX revealed that this capacity differed. It was found that alpha 1-PDX exerts its effect by interfering with the formation of syncytia between J-PDX cells infected with gp160 recombinant vaccinia virus, or after infection by HIV-1 and co-culture with uninfected Molt-4 cells. In contrast, when the same experiments were performed with J-pcDNA3 cells, a large number of syncytia was obtained. Analysis of viral proteins by Western blotting and densitometry showed that the inhibition of the cytopathic effect of HIV-1 and viral replication was correlated with the capacity of alpha 1-PDX to interfere with the maturation of gp160 to gp120 and gp41. PMID- 11062062 TI - Thermodynamic mixing of molecular states of the epidermal growth factor receptor modulates macroscopic ligand binding affinity. AB - The epidermal growth factor receptor (EGFr), when expressed on the cell surface, has long been known to display two distinct affinities for epidermal growth factor (EGF) binding. In addition, the treatment of cells expressing the EGFr with phorbol esters has been shown to cause a loss of the high-affinity binding capacity of the receptor. In the present study, point mutations that alter acidic or phosphorylation sites have been made in an intracellular domain near Tyr-992 (residues 988-992) of the EGFr. Equilibrium (125)I-EGF binding studies demonstrate that the conversion of Tyr-992 into glutamate induces a 4-fold decrease in the EGFr apparent low-affinity dissociation constant, whereas the mutation of two acidic residues, Asp-988 and Glu-991, or the conversion of Tyr 992 into phenylalanine does not alter EGFr affinity. Phorbol ester treatment of EGFr-expressing Chinese hamster ovary cells results in a loss of high-affinity binding and an increase in the apparent low-affinity dissociation constant of the receptor, similar to the effect of a truncation mutant in which the C-terminal 190 residues are deleted. These results are examined in the context of a new model for regulation of the affinity of the EGFr for EGF in which a cytosolic particle stabilizes the high-affinity conformation of the EGFr and a rapid equilibrium exists between EGFr high-affinity and low-affinity conformations. This model demonstrates that the macroscopic affinities of the EGFr can differ from the affinities of individual EGFr molecules and provides a theoretical framework whereby the measured affinities of the EGFr are modulated by intracellular interactions. PMID- 11062063 TI - 3'-untranslated regions of oxidative phosphorylation mRNAs function in vivo as enhancers of translation. AB - Recent findings have indicated that the 3'-untranslated region (3'-UTR) of the mRNA encoding the beta-catalytic subunit of the mitochondrial H(+)-ATP synthase has an in vitro translation-enhancing activity (TEA) [Izquierdo and Cuezva, Mol. Cell. Biol. (1997) 17, 5255-5268; Izquierdo and Cuezva, Biochem. J. (2000) 346, 849-855]. In the present work, we have expressed chimaeric plasmids that encode mRNA variants of green fluorescent protein in normal rat kidney and liver clone 9 cells to determine whether the 3'-UTRs of nuclear-encoded mRNAs involved in the biogenesis of mitochondria have an intrinsic TEA. TEA is found in the 3'-UTR of the mRNAs encoding the alpha- and beta-subunits of the rat H(+)-ATP synthase complex, as well as in subunit IV of cytochrome c oxidase. No TEA is present in the 3'-UTR of the somatic mRNA encoding rat mitochondrial transcription factor A. Interestingly, the TEA of the 3'-UTR of mRNAs of oxidative phosphorylation is different, depending upon the cell type analysed. These data provide the first in vivo evidence of a novel cell-specific mechanism for the control of the translation of mRNAs required in mitochondrial function. PMID- 11062064 TI - In vitro membrane-inserted conformation of the cytochrome b(5) tail. AB - The cytochrome b(5) tail is a 43-residue membrane-embedded domain that is responsible for anchoring the catalytic domain of cytochrome b(5) to the endoplasmic reticulum membrane. Different models for the structure of the membrane domain of cytochrome b(5) have been proposed, including a helical hairpin and a single transmembrane helix. In the present study, CD spectroscopy was used to investigate the conformation of the tail in different environments, and as a function of temperature, with the goal of understanding the nature of the membrane-bound conformation. Whereas residue property profiling indicates that bending of a helix in the middle of the peptide might be possible, the experimental results in small unilamellar vesicles and lysophosphatidylcholine micelles are more consistent with a single transmembrane helix. Furthermore, although there is evidence for some refolding of the polypeptide with temperature, this is not consistent with a hairpin-to-transmembrane transition. Rather, it appears to represent an increase in helical content in fluid lipid environments, perhaps involving residues at the ends of the transmembrane segment. PMID- 11062065 TI - Macrophage cholesteryl ester hydrolases and hormone-sensitive lipase prefer specifically oxidized cholesteryl esters as substrates over their non-oxidized counterparts. AB - The oxidative modification of low-density lipoprotein (LDL) has been implicated as a pro-atherogenic process in the pathogenesis of atherosclerosis. Macrophages rapidly take up oxidized LDL via scavenger-receptor-mediated pathways and thereby develop into lipid-laden foam cells. The uptake mechanism has been studied extensively and several types of scavenger receptors have been identified. In contrast, the intracellular fate of oxidized LDL lipids is less well investigated. We studied the degradation of specifically oxidized cholesteryl esters by murine macrophages using an HPLC-based assay, and found that oxidized substrates are hydrolysed preferentially from a 1:1 molar mixture of oxidized and non-oxidized cholesteryl esters. This effect was observed at both neutral and acidic pH. Similar results were obtained with lysates of human monocytes and with pure recombinant human hormone-sensitive lipase. These data suggest that the intracellular oxidation of cholesteryl esters may facilitate intracellular cholesteryl ester hydrolysis, and thus may represent an anti-atherogenic process. PMID- 11062066 TI - Lysophosphatidic acid prevents apoptosis in fibroblasts via G(i)-protein-mediated activation of mitogen-activated protein kinase. AB - Lysophosphatidic acid (LPA) is a naturally occurring phospholipid with multiple biological functions. In the present study, we demonstrate that, besides its mitogenic activity, LPA is a potent survival factor, preventing serum-deprivation induced apoptosis in fibroblasts and other cell types. Both the proliferative effect and survival activity of LPA are sensitive to the action of pertussis toxin (PTX), indicating that both processes are mediated by G(i) protein(s). We therefore focused on the role of G(i)-protein-mediated signalling events in the promotion of cell survival by LPA. In addition to activation of mitogen-activated protein kinase (MAPK), LPA stimulates a modest PTX-sensitive phosphorylation/activation of the serine/threonine kinase Akt, a survival mediator downstream of phosphoinositide 3-kinase (PI3K). Inhibition of PI3K with LY 294002 or wortmannin resulted in a marked inhibition of LPA-induced DNA synthesis, and yet the survival activity of LPA decreased by only 20-30%, suggesting a limited input of the PI3K-Akt cascade in LPA-induced cell survival. In contrast, inhibition of MAPK activation by the MEK-1 inhibitor, PD 98059, blocked both the proliferative and survival effects of LPA. These results indicate that LPA promotes cell survival largely via G(i)-protein-mediated activation of ERK1/ERK2, or other PD 98059-sensitive member(s) of the MAPK family. PMID- 11062067 TI - Synergistic activation of stress-activated protein kinase 1/c-Jun N-terminal kinase (SAPK1/JNK) isoforms by mitogen-activated protein kinase kinase 4 (MKK4) and MKK7. AB - Stress-activated protein kinase 1 (SAPK1), also called c-Jun N-terminal kinase (JNK), becomes activated in vivo in response to pro-inflammatory cytokines or cellular stresses. Its full activation requires the phosphorylation of a threonine and a tyrosine residue in a Thr-Pro-Tyr motif, which can be catalysed by the protein kinases mitogen-activated protein kinase kinase (MKK)4 and MKK7. Here we report that MKK4 shows a striking preference for the tyrosine residue (Tyr-185), and MKK7 a striking preference for the threonine residue (Thr-183) in three SAPK1/JNK1 isoforms tested (JNK1 alpha 1, JNK2 alpha 2 and JNK3 alpha 1). For this reason, MKK4 and MKK7 together produce a synergistic increase in the activity of each SAPK1/JNK isoform in vitro. The MKK7 beta variant, which is several hundred-fold more efficient in activating all three SAPK1/JNK isoforms than is MKK7 alpha', is equally specific for Thr-183. MKK7 also phosphorylates JNK2 alpha 2 at Thr-404 and Ser-407 in vitro, Ser-407 being phosphorylated much more rapidly than Thr-183 in vitro. Thr-404/Ser-407 are phosphorylated in unstimulated human KB cells and HEK-293 cells, and phosphorylation is increased in response to an osmotic stress (0.5 M sorbitol). However, in contrast with Thr 183 and Tyr-185, the phosphorylation of Thr-404 and Ser-407 is not increased in response to other agonists that activate MKK7 and SAPK1/JNK, suggesting that phosphorylation of these residues is catalysed by another protein kinase, such as CK2, which also phosphorylates Thr-404 and Ser-407 in vitro. MKK3, MKK4 and MKK6 all show a strong preference for phosphorylation of the tyrosine residue of the Thr-Gly-Tyr motifs in their known substrates SAPK2a/p38, SAPK3/p38 gamma and SAPK4/p38 delta. MKK7 also phosphorylates SAPK2a/p38 at a low rate (but not SAPK3/p38 gamma or SAPK4/p38 delta), and phosphorylation occurs exclusively at the tyrosine residue, demonstrating that MKK7 is intrinsically a 'dual-specific' protein kinase. PMID- 11062068 TI - Catalytic activation of mitogen-activated protein (MAP) kinase phosphatase-1 by binding to p38 MAP kinase: critical role of the p38 C-terminal domain in its negative regulation. AB - Mitogen-activated protein (MAP) kinase phosphatase-1 (MKP-1) is the archetypal member of the dual-specificity protein phosphatase family, the expression of which can be rapidly induced by a variety of growth factors and cellular stress. Since MKP-1 protein localizes in the nucleus, it has been suggested to play an important role in the feedback control of MAP kinase-regulated gene transcription. Recently it has been demonstrated that the interaction of several cytosolic MAP kinase phosphatases with MAP kinases can trigger the catalytic activation of the phosphatases. It is unclear whether such a regulatory mechanism can apply to nuclear MAP kinase phosphatases and serve as an additional apparatus for the feedback control of MAP kinase-mediated gene expression. Here we have shown that MKP-1 associates directly with p38 MAP kinase both in vivo and in vitro, and that this interaction enhances the catalytic activity of MKP-1. The point mutation Asp-316-->Asn in the C-terminus of p38, analogous to the ERK2 (extracellular-signal-regulated kinase 2) sevenmaker mutation, dramatically decreases its binding to MKP-1 and substantially compromises its stimulatory effect on the catalytic activity of this phosphatase. Consistent with its defective interaction with MKP-1, this p38 mutant also displays greater resistance to dephosphorylation by the phosphatase. Our studies provide the first example of catalytic activation of a nuclear MAP kinase phosphatase through direct binding to a MAP kinase, suggesting that such a regulatory mechanism may play an important role in the feedback control of MAP kinase signalling in the nuclear compartment. PMID- 11062069 TI - Identification of Rab6 as an N-ethylmaleimide-sensitive fusion protein-binding protein. AB - In this study we show the interaction of N-ethylmaleimide-sensitive fusion protein (NSF) with a small GTP-binding protein, Rab6. NSF is an ATPase involved in the vesicular transport within eukaryotic cells. Using the yeast two-hybrid system, we have isolated new NSF-binding proteins from the rat lung cDNA library. One of them was Rab6, which is involved in the vesicular transport within the Golgi and trans-Golgi network as a Ras-like GTPase. We demonstrated that the N terminal domain of NSF interacted with the C-terminal domain of Rab6, and these proteins were co-immunoprecipitated from the rat brain extract. This interaction was maintained preferentially in the presence of hydrolysable ATP. Recombinant NSF-His(6) can also bind to C-terminal Rab6-glutathione S-transferase under the conditions to allow the ATP hydrolysis. Surprisingly, Rab6 stimulates the ATPase activity of NSF by approx. 2-fold as does alpha-soluble NSF attachment protein receptor. Anti-Rab6 polyclonal antibodies significantly inhibited the Rab6 stimulated ATPase activity of NSF. Furthermore, we found that Rab3 and Rab4 can also associate with NSF and stimulate its ATPase activity. Taken together, we propose a model in which Rab can form an ATP hydrolysis-regulated complex with NSF, and function as a signalling molecule to deliver the signal of vesicle fusion through the interaction with NSF. PMID- 11062070 TI - Divergence in the anti-apoptotic signalling pathways used by nerve growth factor and basic fibroblast growth factor (bFGF) in PC12 cells: rescue by bFGF involves protein kinase C delta. AB - The mechanisms whereby nerve growth factor (NGF) and basic fibroblast growth factor (bFGF) block apoptosis in serum-deprived PC12 cells were investigated. NGF, but not bFGF, strongly activated Akt/protein kinase B, a downstream effector of phosphoinositide (phosphatidylinositol) 3-kinase (PI 3-kinase). In addition, inhibition of PI 3-kinase by LY294002 partially blocked inhibition of apoptosis by NGF, but not that by bFGF, suggesting divergence in NGF and bFGF anti apoptotic signalling pathways. Both growth factors strongly activated mitogen activated protein (MAP) kinases, but blockade of signalling through this pathway, either by the expression of dominant-negative Ras or by treatment with the MAP kinase/ERK kinase (MEK) inhibitor U0126, partially inhibited only bFGF, but not NGF, anti-apoptotic signalling. Use of isoform-specific protein kinase C (PKC) inhibitors such as bisindoylmaleimide-I and Go 6983 suggested that PKC delta is a likely component of bFGF trophic signalling. A role for PKC delta was confirmed in PC12 cells expressing a dominant-negative PKCdelta fragment, in which reversal of apoptosis by bFGF was partially blocked. The PKC delta signal was not mediated by the MAP kinase cascade, as bFGF activation of this pathway was not affected in cells expressing the dominant-negative PKC delta fragment. Full inhibition of bFGF anti-apoptotic signalling occurred when both the PKCdelta and Ras/MAP kinase pathways were inhibited. Together, these data demonstrate that inhibition of apoptosis by bFGF in PC12 cells operates differently from that mediated by NGF, requiring the addition of signals from both the Ras/MAP kinase and PKC signalling pathways. PMID- 11062071 TI - Lentil seed aquaporins form a hetero-oligomer which is phosphorylated by a Mg(2+) dependent and Ca(2+)-regulated kinase. AB - In plants, aquaporins regulate the water flow through membranes during growth, development and stress responses. We have isolated two isoforms of the aquaporin family from the protein-storage vacuoles of lentil (Lens culinaris Med.) seeds. Chemical cross-linking experiments showed that both isoforms belong to the same oligomer in the membrane and are phosphorylated by a membrane-bound protein kinase. We assigned the kinase activity to a 52 kDa protein that is magnesium dependent and calcium-regulated. PMID- 11062072 TI - Bovine prion protein as a modulator of protein kinase CK2. AB - On the basis of far-Western blot and plasmon resonance (BIAcore) experiments, we show here that recombinant bovine prion protein (bPrP) (25-242) strongly interacts with the catalytic alpha/alpha' subunits of protein kinase CK2 (also termed 'casein kinase 2'). This association leads to increased phosphotransferase activity of CK2alpha, tested on calmodulin or specific peptides as substrate. We also show that bPrP counteracts the inhibition of calmodulin phosphorylation promoted by the regulatory beta subunits of CK2. A truncated form of bPrP encompassing the C-terminal domain (residues 105-242) interacts with CK2 but does not affect its catalytic activity. The opposite is found with the N-terminal fragment of bPrP (residues 25-116), although the stimulation of catalysis is less efficient than with full-size bPrP. These results disclose the potential of the PrP to modulate the activity of CK2, a pleiotropic protein kinase that is particularly abundant in the brain. PMID- 11062073 TI - Angiotensin II stimulates cyclic ADP-ribose formation in neonatal rat cardiac myocytes. AB - To examine the role of cyclic ADP-ribose (cADP-ribose) as a second messenger downstream of angiotensin II (Ang II) receptor activation in the heart, ADP ribosyl cyclase activity was measured in a crude membrane fraction of ventricular myocytes. Ang II at 10-100 nM increased ADP-ribosyl cyclase activity by 40-90% in the ventricular muscle of neonatal (2-4-day-old) rats, but not in fetal or adult hearts. This increase was inhibited by the Ang II antipeptide. Stimulation of ADP ribosyl cyclase was reproduced by GTP and guanosine 5'-[gamma-thio]triphosphate, and prevented by guanosine 5'-[beta-thio]diphosphate. Prior treatment of the rats with cholera toxin A and B subunits also blocked the Ang II-induced activation. The density of Ang II receptors detected as [(3)H]Ang II binding was higher in neonatal than adult rats. These results demonstrate the existence of a signalling pathway from Ang II receptors to membrane-bound ADP-ribosyl cyclase in the ventricular muscle cell and suggest that the Ang II-induced increase in cADP ribose synthesis is involved in the regulation of cardiac function and development. PMID- 11062074 TI - Dominance of G(s) in doubly G(s)/G(i)-coupled chimaeric A(1)/A(2A) adenosine receptors in HEK-293 cells. AB - A(1) adenosine receptors inhibit adenylate cyclase by activating G(i)/G(o), whereas A(2A) receptors activate G(s). We examined how regions of A(1) and A(2A) receptors regulate coupling to G-proteins by constructing chimaeras in which the third intracellular loops (3ICL or L) and/or the C-termini (or T) were switched. Pertussis toxin (PTX) was used in membrane radioligand binding assays to calculate the fraction of recombinant receptors coupled to G(i)/G(o) and in whole cells to differentially influence agonist-stimulated cAMP accumulation. Switching A(1)/A(2A) 3ICL domains results in receptors that maintain binding selectivity for ligands but are doubly coupled. Receptor chimaeras with an A(1) 3ICL sequence (A(2A)/A(1)L or A(2A)/A(1)LT) respond to agonist stimulation with elevated cAMP despite being coupled predominantly to G(i)/G(o). These chimaeras have basal cAMP levels lower than those of wild-type A(2A) receptors, similar to wild-type A(1) receptors. The A(1) C-terminus modulates the coupling of receptors with A(1) 3ICL such that A(2A)/A(1)LT is better coupled to G(i)/G(o) than A(2A)/A(1)L. The C terminus has little impact on coupling to receptors containing A(2A) 3ICL sequence. Our results show that the C-terminus sequence selectively facilitates coupling to G(i)/G(o) mediated by A(1) 3ICL and not by other intracellular domains that favour G(i) coupling. The C-terminus sequence has little or no effect on coupling to G(s). For doubly G(s)/G(i)-coupled adenosine receptors in HEK-293 cells, G(s)-mediated stimulation predominates over G(i)/G(o)-mediated inhibition of adenylate cyclase. We discuss the signalling consequences of simultaneously activating opposing G-proteins within single cells. PMID- 11062075 TI - Regulation by glucagon (cAMP) and insulin of the promoter of the human phosphoenolpyruvate carboxykinase gene (cytosolic) in cultured rat hepatocytes and in human hepatoblastoma cells. AB - A promoter fragment (-457 to +65) of the human cytosolic phosphoenolpyruvate carboxykinase gene, which by analogy to the rat promoter contains regulatory regions conferring glucagon (cAMP) and insulin responsiveness to the phosphoenolpyruvate carboxykinase gene, was cloned into a luciferase expression vector and transfected into cultured rat hepatocytes and human hepatoblastoma cells (HepG2) to study the regulation of the transgene by glucagon (cAMP) and insulin. A reporter gene that contained the rat promoter sequence from -493 to +33 was used for comparison. In cultured rat hepatocytes glucagon and its second messenger cAMP increased luciferase expression 4-6-fold over basal levels. Insulin reduced this effect by 40-70%. Luciferase expression was also stimulated by the combination of dexamethasone and cAMP in HepG2 cells and this effect was inhibited by insulin. The phosphoinositide 3-kinase (PI 3-kinase) inhibitor, wortmannin, abolished this action of insulin in cultured rat hepatocytes. The results show that the promoter of the human phosphoenolpyruvate carboxykinase gene mediates the stimulatory action of glucagon and its second messenger cAMP. The inhibitory action of insulin was exerted through the PI 3-kinase pathway in cultured rat hepatocytes. PMID- 11062076 TI - Peroxynitrite activates the phosphoinositide 3-kinase/Akt pathway in human skin primary fibroblasts. AB - Peroxynitrite is a potent oxidizing and nitrating species formed in a diffusion limited reaction between nitrogen monoxide and superoxide. It induces apoptosis through unknown mechanisms and is believed to interfere with receptor tyrosine kinase signalling through nitration of tyrosine residues. One pathway emanating from receptor tyrosine kinases is that leading to activation of the anti apoptotic kinase Akt. In the present study we provide evidence that peroxynitrite, administered to cells using two different delivery systems, results in the dose- and time-dependent activation of Akt. Akt activation is rapid and followed by phosphorylation of glycogen synthase kinase-3, an established substrate of Akt. Akt activation is inhibited in the presence of the phosphoinositide 3-kinase (PI-3K) inhibitors wortmannin and LY294002, and by treatment with the platelet-derived growth factor (PDGF) receptor (PDGFR) inhibitor AG1295, indicating a requirement for PDGFR and PI-3K in mediating peroxynitrite-induced Akt activation. Accordingly, the PDGFR-A and PDGFR-B isoforms were shown to undergo rapid tyrosine phosphorylation on treatment with peroxynitrite. Prior exposure of cells to peroxynitrite interferes with PDGF induced Akt phosphorylation. Our findings suggest that Akt activation occurs as an acute response to peroxynitrite treatment and could play an important role in influencing cell survival and/or alter the cellular response to other growth regulatory signals. PMID- 11062077 TI - Dietary P(i) deprivation in rats affects liver cAMP, glycogen, key steps of gluconeogenesis and glucose production. AB - We previously reported [Xie, Li, Mechin and van de Werve (1999) Biochem. J. 343, 393-396] that dietary phosphate deprivation for 2 days up-regulated both the catalytic subunit and the putative glucose-6-phosphate translocase of the rat liver microsomal glucose-6-phosphatase system, suggesting that increased hepatic glucose production might be responsible for the frequent clinical association of hypophosphataemia and glucose intolerance. We now show that liver cAMP was increased in rats fed with a diet deficient in P(i) compared with rats fed with a control diet. Accordingly, in the P(i)-deficient group pyruvate kinase was inactivated, the concentration of phosphoenolpyruvate was increased and fructose 2, 6-bisphosphate concentration was decreased. Phosphoenolpyruvate carboxykinase activity was marginally increased and glucokinase activity was unchanged by P(i) deprivation. The liver glycogen concentration decreased in the P(i)-deficient group. In the fed state, plasma glucose concentration was increased and plasma P(i) and insulin concentrations were substantially decreased in the P(i) deficient group. All of these changes, except decreased plasma P(i), were cancelled in the overnight fasted P(i)-deficient group. In the fasted P(i) deficient group, immediately after a glucose bolus, the plasma glucose level was elevated and the inhibition of endogenous glucose production was decreased. However, this mild glucose intolerance was not sufficient to affect the rate of fall of the glucose level after the glucose bolus. Taken together, these changes are compatible with a stimulation of liver gluconeogenesis and glycogenolysis by the P(i)-deficient diet and further indicate that the liver might contribute to impaired glucose homeostasis in P(i)-deficient states. PMID- 11062078 TI - Endocytic uptake of advanced glycation end products by mouse liver sinusoidal endothelial cells is mediated by a scavenger receptor distinct from the macrophage scavenger receptor class A. AB - Previous studies with peritoneal macrophages obtained from macrophage scavenger receptor class A (MSR-A) knock-out mice showed that the endocytic uptake of advanced glycation end products (AGE) by macrophages was mediated mainly by MSR A. However, it is controversial whether the endocytic uptake of intravenously injected AGE proteins by liver sinusoidal endothelial cells (LECs) is similarly explained by receptor-mediated endocytosis via MSR-A. The present study was conducted to compare the capacity to endocytose AGE proteins in LECs and peritoneal macrophages obtained from MSR-A knock-out and littermate wild-type mice. The endocytic degradation capacity of MSR-A knock-out LECs for AGE-BSA was indistinguishable from that of wild-type LECs, whereas that of MSR-A knock-out peritoneal macrophages for AGE-BSA was decreased to 30% of that in wild-type cells. Similarly, the endocytic degradation of MSR-A knock-out LECs for acetylated low-density lipoprotein (acetyl-LDL) did not differ from that of wild type LECs, whereas the endocytic degradation of acetyl-LDL by MSR-A knock-out peritoneal macrophages was less than 20% of that in wild-type cells. Furthermore, formaldehyde-treated serum albumin (f-Alb), a ligand known to undergo scavenger receptor-mediated endocytosis by LECs, was effectively taken up by MSR-A knock out LECs at a capacity that did not differ from that of wild-type LECs. Moreover, the endocytic uptake of AGE-BSA by LECs was effectively competed for by unlabelled f-Alb or acetyl-LDL. These results indicate that the scavenger receptor ligands AGE proteins, acetyl-LDL and f-Alb are endocytosed by LECs through a non-MSR-A pathway. PMID- 11062079 TI - [Nuclear oncology 2000: the end of individualized professionalism]. PMID- 11062080 TI - [Treatment with (131)I of toxic multinodular goiter. Assessment of a fixed dose protocol]. AB - OBJECTIVE: Present the results of treatment with 131I in a cohort of 119 patients diagnosed of toxic multinodular goiter (TMG) from a specific geographic area who had been treated with fixed doses of isotopes (555 MBq). The indication for the administration of the 131I was hyperthyroidism correction in all of the cases. MATERIAL AND METHODS: 111 of the 119 patients treated were women (93%) and 8 were men (7%) and their ages ranged from 38 to 88 years. The follow-up periods were always greater than one year (1.78 +/- 1.05 years). Treatment with thyroid medication was discontinued in all of the patients at least 7 days prior to the administration of the isotope. RESULTS: 93 (78.1%) of the 119 patients remained euthyroidic, 10 (8.4%) presented hypothyroidism, 11 (9.2%) hyperthyroidism and 5 (4.2%) had sub-clinical levels of hyperthyroidism at the end of the follow-up period. No statistically significant relationship were found between hypothyroidism, gender and previous administration of thyroid medication. CONCLUSIONS: The use of 131I is an alternative to the treatment of TMG produced hyperthyroidism. The fixed doses of 555 Mbq that we propose are comfortable, make it unnecessary for the patients to travel in extensive geographic areas and produce a low rate of hypothyroidism. In our opinion, the differences in the iodine content of the diets that continue to exist are one of the factors that could influence in the differences in results observed in the different series, although other factors, such as the size of the goiter and the heterogeneity of the radioisotope uptake can contribute to the variations found in our patients. PMID- 11062081 TI - [Perfusion SPECT with (99m)Tc-HMPAO in type I diabetics with no background of central neurologic symptoms. A study of activation with acetazolamide]. AB - OBJECTIVE: This study aimed to assess if activation with acetazolamide increases the diagnostic capacity of baseline SPECT with (99m)Tc-HMPAO in the study of brain perfusion in type I diabetic patients with no history of neurological symptoms. MATERIAL AND METHODS: A baseline SPECT was carried out in 11 diabetes mellitus type I patients with no neurological symptoms with 555 MBq of (99m)Tc HMPAO; 1 g of acetazolamide was administered during the examination and a second SPECT was obtained 20' later with the same methodology used in the baseline SPECT. The images were visually analyzed. The post-acetazolamide studies were analyzed with (CBS) and without (WBS) baseline image subtraction and both methods were compared. RESULTS: The baseline SPECT showed 48 hypoperfused cortical areas. The post-acetazolamide SPECT analyzed without baseline image subtraction detected 14 new hypoperfused areas and those analyzed with it detected 26 areas. 69% of the baseline hypoperfused areas were hyporeactive in the WBS analysis and 54% in the CBS analysis. CONCLUSION: The perfusion SPECT with acetazolamide improves the diagnostic capacity of the baseline perfusion (99m)Tc-HMPAO SPECT, and makes it possible to classify the abnormalities as metabolic or vascular, with a preference for the post-acetazolamide CBS imaging analysis. PMID- 11062082 TI - [Usefulness of quantification and visual analysis of the uptake of (99m)Tc-MIBI in the diagnosis of mammary lesions]. AB - OBJECTIVE: Scintigraphy studies with (99m)Tc-MIBI are widely used in the diagnosis of non-invasive breast cancer and their results have been verified by many studies. However, the scintigraphic technique produces erroneous false negative and positive results. This study aims to verify how the different characteristics of the uptake of (99m)Tc-MIBI (intensity, size, morphology, etc.) can help to increase sensitivity and specificity of breast scintigraphy. MATERIALS AND METHODS: We have studied 201 patients (84 breast cancer and 117 benign lesions). All of them underwent a breast scintigraphy with (99m)Tc-MIBI and anatomopathological study. A semiquantitative analysis (by T/F indexes) and visual study were performed in the lesions that presented radiodrug uptake. RESULTS: Eleven false positive results and 8 false negative results were obtained in the study of the 201 patients. The analysis of the T/F indexes did not demonstrate any significant differences between the benign and malignant lesions (p>0,05). There is a statistically significant correlation (p<0.05) between the tracer uptake morphology and the AP diagnosis and between the localization of the lesion in a breast quadrant and the uptake intensity. CONCLUSIONS: Quantification of (99m)Tc-MIBI uptake by T/F indexes does not provide any conclusive data on the lesion's malignancy, however, a more detailed analysis of the characteristics of the tracer uptake would permit us to make the correct diagnosis and would reduce the false positive and negative results of this technique. PMID- 11062083 TI - [Plasma levels of insulin and leptin in patients with morbid obesity and anorexia nervosa after weight loss or gain, respectively]. AB - The present study was conducted in order to analyze the relationship existing between leptin and insulin levels in massive weight loss and weight recovery. Thirteen patients with severe obesity, 14 patients with anorexia nervosa and 13 healthy control subjects were studied. The patients with severe obesity underwent a vertical banded gastroplasty followed by an 800 kcal/day diet for 12 weeks. They were evaluated prior to (body mass index [BMI] 51.2 +/- 8.8 Kg/m2) and after drastic weight loss (BMI 40.6 +/- 6.7 Kg/m2). Patients with anorexia nervosa were treated exclusively with nutritional therapy during 12 weeks, and they were evaluated at their lowest weight status (BMI 16.2 +/- 2.2 Kg/m2) and after weight recovery (BMI 17.9 +/- 2.3 Kg/m2). The BMI of the normal subjects was in the normal range of 20 to 27 Kg/m2 (average 22.8 +/- 2.6 Kg/m2). BMI, percentage of body fat, waist circumference, and serum levels of leptin, insulin, and C-peptide were determined in each patient and normal subject. In severely obese patients, serum leptin and insulin decreased significantly after drastic weight reduction (leptin: from 51.8 +/- 22.3 to 23.7 +/- 10.2 ng/ml; insulin: from 27.1 +/- 13.3 to 17.2 +/- 7.2 mU/ml). In patients with anorexia nervosa, the mean serum leptin levels were significantly higher after weight recovery (5.5 +/- 3.2 vs 7.6 +/- 6 ng/ml). Serum leptin in the severe obesity group correlated positively with BMI, percentage body fat and waist circumference before and after weight loss. In those patients suffering from anorexia nervosa, serum leptin correlated positively with the BMI, percentage of body fat, and waist circumference in the low weight state and after weight recovery. In addition, their serum insulin correlated with BMI and waist circumference after weight recovery. These data reveal that serum leptin concentration correlates significantly with the BMI and body fat content 1) in subjects with a range of weight and caloric intake, 2) in obese patients after drastic weight loss; 3) in anorexic patients after weight gain; and that hyper- or normoinsulinemia do not seem to have any influence on the leptin changes caused by weight loss or gain. PMID- 11062084 TI - [Locating the sentinel node in breast cancer by gamma probe and staining agent. Preliminary study]. AB - OBJECTIVE: The sentinel node biopsy appears to offer an alternative to routine axillary lymph node dissection for staging patients with breast cancer. Various techniques have been studied for identifying the sentinel node, using vital blue or radioactive colloid. This study aimed to evaluate our preliminary results with these techniques. MATERIAL AND METHODS: In this work carried out in the Valencian Institute of Oncology, 21 women with T1- T2 breast cancer with an average age of 52 years (range: 42-73) underwent the sentinel node biopsy, which was immediately followed by standard axillary dissection. Both blue dye and radioisotope were used to identify the sentinel node. The radioactive axillary of sentinel node was localized by the gamma probe. RESULTS: The sentinel node was successfully identified by lymph node scintigraphy in 100%. It was localized by blue dye in 33% and by combination of blue dye and isotope in 95%. Of the 21 patients in this study in whom sentinel nodes were identified, 7 (35%) were histologically positive; in 6 cases, the sentinel was the only site of the metastases (86%). The histology of the sentinel node accurately predicted axillary node status in 95% of cases. CONCLUSIONS: The sentinel node biopsy technique is a promising and feasible procedure in patients with clinically T1-T2 N0M0 breast cancer, providing valuable axillary staging information. PMID- 11062085 TI - [Use of SPECT with (99m)Tc-Sestamibi in a patient affected by laryngeal carcinoma and parathyroid adenoma]. AB - We report the case of a patient with a laryngeal carcinoma in whom asymptomatic hyperparathyroidism was also detected during the preoperative work-up. A planar (201)Thallium/(99m)Tc-pertecnetate subtraction scintigraphy was performed in order to locate the suspected parathyroid adenoma. The study showed a single area of increased (201)Thallium uptake just above the thyroid isthmus, likely due to the laryngeal tumor. The scintigraphic study was repeated using (99m)Tc-Sestamibi and (99m)Tc-pertechnetate and employing the SPECT technique. Both SPECT studies made it possible to identify correctly the parathyroid adenoma, located inferiorly and in a posterior position to the lower third of the right thyroid lobe. The laryngeal tumor and parathyroid adenoma could be excised in a single surgery session. This case is of interest due to the rarity of the coexistence of two neck tumors and the clear advantage shown by the SPECT technique with (99m)Tc Sestamibi over the planar technique with 201Thallium. PMID- 11062086 TI - [Detection of bronchopleural fistula by ventilation scintigraphy. Based on one case]. AB - Bronchopleural fistula (BPF) is a rare but extremely serious complication of lung resection surgery. Its diagnosis is often overlooked in the clinical evolution of the patients undergoing surgery due to the subacute presentation of the picture. The procedures used to detect it are either invasive or have low accuracy. We report the case of a patient with lung carcinoma treated by left pneumonectomy in whom BPF was suspected after an episode of coughing and expectoration. The conventional chest x-ray did not show any significant findings while the (99m)Tc DTPA ventilation lung scintigraphy demonstrated bronchopleural communication in a very early phase. PMID- 11062087 TI - Clinical PET in oncology. PMID- 11062088 TI - [Scintigraphy of somatostatin receptors in the diagnosis and staging of neuroendocrine tumors of the digestive tract and pancreas]. PMID- 11062089 TI - [False positives in the detection of Meckel's diverticulum]. PMID- 11062090 TI - [Clinical-biological meaning of progesterone receptor status in 118 estrogen receptor positive infiltrating ductal carcinomas of the breast in post-menopausal women]. PMID- 11062091 TI - [Clinical and biological differences between tetraploid and hypertetraploid infiltrating ductal carcinomas of the breast. Preliminary study]. PMID- 11062092 TI - [Thyroid uptake of iodine]. PMID- 11062094 TI - [News] [In Process Citation] PMID- 11062093 TI - [The Royal Spanish Pharmacopoeia: labeling platelets with (111)indium-oxine]. PMID- 11062095 TI - [Nomenclature, nuclear medicine and PET]. PMID- 11062096 TI - [Usefulness of the quantification of (99M)Tc-MIBI uptake in breast neoplasms in the preoperative assessment of tumor aggressiveness]. AB - (99m)Tc-MIBI has been proposed as an imaging diagnostic method in a large variety of human malignant tumors. At present, the mechanism by which (99m)Tc-MIBI is uptaken and concentrated by the malignant cells is not totally known. Some mammary neoplasms do not show any uptake of (99m)Tc-MIBI. This study aims to determine if there is any correlation between the uptake of (99m)Tc-MIBI by the tumor and the different histopathological parameters involved in tumoral aggressiveness. To do so, we have studied 100 patients with breast cancer. All of them underwent a breast scintimammography with (99m)Tc-MIBI with semiquantitative analysis by means of a tumor-to-background ratio calculated in every projection. After surgery, an experienced pathologist determined tumor size, axillary lymph node metastases, histological grade (Scarff Bloom Richardson) (SCBR), nuclear grade, mitotic index, presence of cellular atypia and estrogen and progesterone receptor expression. RESULTS: A statistically significant correlation (p < 0.005) has been found between tumor-to-background (T/B) ratios of (99m)Tc-MIBI uptake and tumor SCBR histological grade. A correlation between (99m)Tc-MIBI uptake and the mitotic index, cellular atypia and nuclear grade has also been found. No correlation was found in our study with tumor size, hormone receptor expression or axillary lymph node metastases. CONCLUSIONS: (99m)Tc-MIBI uptake in breast cancer is correlated with the tumoral differentiation grade: the smaller the tumoral cellular differentiation (greater aggressiveness), the greater the uptake. On the other hand, no correlation was found between the uptake of (99m)Tc MIBI and the classical pathological parameters that define tumoral aggressiveness, such as size and axillary lymph node metastasis. PMID- 11062097 TI - [Treatment of metastatic bone pain with repeated doses of strontium-89 in patients with prostate neoplasm]. AB - This study has aimed to evaluate the usefulness of repeated treatment with 89Sr in patients with prostate neoplasm and metastatic bone pain. Seventeen patients with partial or complete response after the first dose were retreated with two or more doses (total of 39 doses). The Karnofsky functional status, pain and degree of analgesia were assessed. After the first dose the response was good in 68% of the patients and partial in 32%. After the second dose, the response was good in 62% of the patients, partial in 15% and there was no response in 23% of the cases. The pre-treatment Karnofsky functional status and duration of the effect of 89Sr was lower after the second dose (p = 0.03, p = 0.02), but there were no statistically significant differences in the type of response. In conclusion, re treatment with 89Sr can be administered safely and with a similar response to that achieved after the first dose. PMID- 11062098 TI - [Radioisotopic synoviorthesis in rheumatoid arthritis. A study of 108 cases]. AB - There are several therapeutic alternatives in the local treatment of rheumatoid arthritis (RA): intraarticular corticosteroids, chemical synoviorthesis, surgical synovectomy and synoviorthesis with radioisotopes. We present the results of an observational study on radioisotopic synoviorthesis carried out in the Valencian health care area from January 1989 to May 1997 which included 108 synoviortheses performed in 51 patients. This study aimed to assess the efficacy and safety of radiosynoviorthesis in the treatment of RA which had not responded to conventional treatment. Good and moderate results were obtained in 76.2% of the cases, there being significant differences in the efficacy of synoviorthesis in patients with and without advanced radiographic alterations. There were few side effects (3.7%). We conclude that radiosynoviortesis is a useful and safe therapeutic tool in RA that does not respond to conventional treatment, that it is more effective in large joints with little cartilaginous deterioration and that its repeated use does not decrease the expected therapeutic effect. PMID- 11062099 TI - [Role of the positron emission tomography (PET) in suspected tumor recurrence when there is increased serum tumor markers]. AB - PURPOSE: Evaluate the usefulness of Positron Emission Tomography with 18F Fluorodeoxyglucose (FDG-PET) to detect lesions when tumor recurrences are suspected due to the progressive increase of tumor markers and the study of their extension with negative or non-conclusive morphostructural diagnostic techniques. MATERIAL AND METHODS: Whole body FDG-PET was per-formed in 72 patients with different cancer diseases, 23 of whom had breast, 18 colorectal, 15 thyroid, 6 lung, 3 ovarian cancer and 7 had other types of cancers. Tumoral recurrence was suspected in all of the cases due to the progressive marker increase and negative or non-conclusive conventional studies. RESULTS: FDG-PET detected lesions in 85% patients, 33% of which were confirmed by the end of the study. In 40% of all cases, therapeutics measures were applied, 14% of which consisted in surgery with intention to cure. Sensitivity, specificity and accuracy were 96.4%, 75.6% and 93.9% respectively. CONCLUSIONS: Whole body FDG-PET is an accurate procedure in diagnosis of recurrent tumoral disease in patients with rising tumoral marker levels and with negative conventional morphostructural extension studies, and can make it possible to change the therapeutic approach in some cases. PMID- 11062100 TI - [Post-radiation pneumonitis in a case of Hodgkin's lymphoma assessed with PET-FDG by residual mediastinal mass]. AB - The PET-FDG is a useful method to assess the post-radiotherapy residual masses in supradiaphragmatic lymphoma patients. We present the case of a patient with residual mediastinic mass revealed by CT after the patient had completed treatment for Hodgkin's disease and in whom the PET study demonstrated a typical pattern of post-radiation pneumonitis. When these patients are re-assessed early after radiotherapy, the different tracer uptake patterns should be taken into account in order to identify the existence of radiotherapy sequela and avoid false positive results. PMID- 11062101 TI - [Subperiosteal abscess as a complication of osteomyelitis: usefulness of bone scintigraphy]. AB - We present the case of an unweaned baby girl with acute femoral osteomyelitis. The bone scintigraphy showed diffuse photopenia of the femoral diaphysis. The final diagnosis revealed diaphyseal osteomyelitis complicated by the presence of a subperiostic abscess. PMID- 11062102 TI - [Value of nuclear medicine tests in oncology]. PMID- 11062103 TI - [Scintigraphy with (99m)Tc-MDP in rhabdomyolysis]. PMID- 11062104 TI - [Clinicobiological study of membrane hyaluronic acid index /CD44v5 (AHm/CD44v5) in patients with infiltrating ductal breast carcinoma]. PMID- 11062106 TI - [News] [In Process Citation] PMID- 11062105 TI - [Carcinoembryonic antigen concentrations in breast cystic fluids increase in type II fluids regardless of whether there are multiple cysts or one cyst. A study of 152 cases]. PMID- 11062107 TI - [Clinical management. Something more than a fashion]. PMID- 11062108 TI - [Lympho-gammagraphy and study of sentinel node in cutaneous melanoma]. AB - OBJECTIVE: To evaluate the role of selective sentinel lymph node dissection to reduce the number of unnecessary lymphadenectomies in patients with intermediate risk of melanoma and without clinical evidence of regional node and distant metastases. MATERIAL AND METHOD: We studied 65 patients with stage I and II melanoma. Only vital blue dye mapping was carried out in nine patients and combined lymphatic mapping with both blue dye and lymphoscintigraphy was used in the remaining 56 patients. RESULTS: The sentinel node was identified in 63 of 65 patients (97%). Only eighth patients (12%) were found to have metastatic melanoma cells in their sentinel node. CONCLUSIONS: Our findings confirm that the intraoperative lymphatic mapping of the sentinel node using both blue dye and radiodetection is an appropriate and simple technique for selecting patients who are more likely to benefit from lymph node dissection. PMID- 11062109 TI - [Results of myocardial scintigraphy with 99mTc-tetrofosmin and dipyridamole administration in patients diagnosed of microvascular angina]. AB - The objective of the study was to evaluate the results of the myocardial scintigraphy with 99mTc-tetrofosmin (Tc-Tf) and pharmacological stimulation with dipyridamole in patients diagnosed of microvascular angina. The study population is made up of 50 patients discharged with the diagnosis of microvascular angina who had undergone Tc-Tf-dipyridamole. Clinical and electrical positivity appeared in 40% and 12% of the patients, respectively. Myocardial perfusion defects were found in 35 patients (70%), and were reversible in 21 (60%), fixed in 11 (21%), and combined in 3 (9%). Abnormalities were inferior, anteroseptal and lateral in 21, 18, and 2 patients, respectively. Patients with a positive exercise treadmill test, compared with those with a negative one, had more frequent perfusion abnormalities (91% vs 50%, p = 0.0327) and myocardial ischemia (64% vs 20%, p = 0.392). Women, in comparison with men, had angina (56% vs 22%, p = 0.013), and anteroseptal perfusion abnormalities (26% vs 4%, p = 0.028) more frequently. On the contrary, men had inferior perfusion abnormalities more frequently (57% vs 30%, p = 0.057). Thus, Tc-Tf-dipyridamole shows perfusion abnormalities in 70% of patients with microvascular angina (91% in patients with a positive exercise treadmill test). Scintigraphic pattern may be partially conditioned by gender in these patients. PMID- 11062110 TI - [99m Tc-MIBI scintigraphy compared to mammography in the diagnosis of breast cancer in dense, operated and young women breasts]. AB - Breast scintimammography with 99mTc-MIBI has proven to be a useful complement to mammography in the diagnosis of breast cancer in the female population. Although the mammography, along with a physical examination, is the backbone of breast cancer diagnosis, there are groups of patients in whom the mammography has an even lower specificity. OBJECTIVE: Our study has aimed to assess the usefulness of breast 99mTc-MIBI scintimammography in those situations in which the mammography was indeterminate, such as, in dense breasts, young females or breasts with architectural distortion after surgery or radiation therapy. MATERIALS AND METHODS: We studied 109 females with mammographically dense breasts, 8 young females under 30 and 24 patients who had undergone previous surgery or radiation therapy. All cases were studied to rule out breast cancer. Final diagnosis was established with excisional biopsy. RESULTS: In dense breasts MIBI scintimammography sensitivity was 88% and the mammography one 81%. MIBI scintimammography specificity was 90% and the mammography 28%. In young females MIBI scintimammography sensitivity was 100% and the mammography 50%, MIBI scintimammography specificity 100% and the mammography 20%. In previous surgery, MIBI scintimammography sensitivity was 80% and the mammography 80%, MIBI scintimammography specificity 100% and the mammography 42%. CONCLUSION: Breast scintimammography with 99mTc-MIBI is an excellent diagnostic technique with high specificity. Undoubtedly it is complementary to mammography in those cases where mammography has major limitations such as dense breasts, young females and breasts with severe scarring after surgery or radiation therapy. PMID- 11062111 TI - [Expression of the adhesion molecule CD44v6 in infiltrating ductal carcinomas of the breast is associated with hormone dependence. Our experience with 168 cases]. AB - In order to investigate the possible hormone-dependence of CD44v6 in human breast cancer, we assayed the concentrations of this isoform in the membrane fraction of 168 invasive ductal carcinomas (IDC) and in 26 normal breast tissue samples, 18 fibradenomas (FAD), 3 fibrocystic disease specimens (FD), 7 mucinous carcinomas and 4 medullary carcinomas using the ELISA method. The results were compared with those of the estrogen (ER) and progesterone (PR) receptors, pS2, tissue type plasminogen activator (t-PA), cathepsin D, epidermal growth factor receptor (EGFR) and c-erbB2/neu oncoprotein concentrations. Menopausal status, size of the tumor in the cases of cancers, axillary lymph node involvement, histologic grade, ploidy, cellular synthesis phase, multifocality and multicentricity were also considered as variables. The cut-off value for CD44v6-positivity was set at 5 ng/mg prt. membrane protein content. 64/138 (38.1%) infiltrating ductal carcinomas scored positive. This was significantly higher than for the normal breast tissue (0/26; p: 0.0001), similar to that seen in the FAD (3/18), fibrocystic disease (0/3), infiltrating mucinous carcinomas (4/7) and lobular (3/15) and significantly lower than for the infiltrating medullary carcinomas (4/4; p: 0.027). There were no significant differences with the other groups of tissues studied. Furthermore, CD44v6-positive IDC showed significantly higher concentrations of ER, PR and cathepsin D and lower (p: 0.051) concentrations of EGFR when compared to their CD44v6-negative counterparts. The significant coexpression of ER, PR and cathepsin D seems to indicate a possible role for hormonal regulation of CD44v6 expression while the role of pS2 and t-PA, estrogen related proteins, was very reduced. PMID- 11062112 TI - [Left distal femoral diaphysis-metaphyseal injury in a 13-year-old patient]. AB - This paper aims to present the usefulness of the different diagnosis imaging methods (anatomical and functional) in the characterization of bone injury. Any data, however insignificant, is justified and should be specified. In this case, the discrepancy between the vascular and pool phases in the bone scintigraphy with 99mTc-MDP reveals revealed a lesion with an intense reaction secondary to the "foreign body effect", which is not necessarily malignant. PMID- 11062113 TI - [Hepatobiliary scintigraphy in a case of acute cholecystitis in childhood]. AB - Acute cholecystitis is an uncommon disease in childhood. Few cases in patients under 10 years of age have been found in literature. An eight-year old male patient with no history of interest who had acute acalculous cholecystitis was reported. Ultrasonographic and isotopic studies led to the diagnosis and control of the patient's evolution. PMID- 11062114 TI - [Scintigraphy with 67Gallium in a case of sarcoidosis]. PMID- 11062115 TI - [Hyperprolactinemia as a cause of breast uptake in 67Ga scintigraphy]. PMID- 11062116 TI - [Lung perfusion scintigraphy. Sociedad Espanola de Medicina Nuclear]. PMID- 11062117 TI - [The sentinel node. Concepts and clinical applications in neoplasms of the breast and melanoma]. PMID- 11062119 TI - [News] PMID- 11062120 TI - Discovery of a transient absorption edge in the X-ray spectrum of GRB 990705. AB - We report the discovery of a transient equivalent hydrogen column density with an absorption edge at approximately 3.8 kiloelectron volts in the spectrum of the prompt x-ray emission of gamma-ray burst (GRB) 990705. This feature can be satisfactorily modeled with a photoelectric absorption by a medium located at a redshift of approximately 0.86 and with an iron abundance of approximately 75 times the solar one. The transient behavior is attributed to the strong ionization produced in the circumburst medium by the GRB photons. The high iron abundance points to the existence of a burst environment enriched by a supernova along the line of sight. The supernova explosion is estimated to have occurred about 10 years before the burst. Our results agree with models in which GRBs originate from the collapse of very massive stars and are preceded by a supernova event. PMID- 11062121 TI - Observation of X-ray lines from a gamma-ray burst (GRB991216): evidence of moving ejecta from the progenitor. AB - We report on the discovery of two emission features observed in the x-ray spectrum of the afterglow of the gamma-ray burst (GRB) of 16 December 1999 by the Chandra X-ray Observatory. These features are identified with the Ly(alpha) line and the narrow recombination continuum by hydrogenic ions of iron at a redshift z = 1.00 +/- 0.02, providing an unambiguous measurement of the distance of a GRB. Line width and intensity imply that the progenitor of the GRB was a massive star system that ejected, before the GRB event, a quantity of iron approximately 0.01 of the mass of the sun at a velocity approximately 0.1 of the speed of light, probably by a supernova explosion. PMID- 11062122 TI - Mode-specific energy disposal in the four-atom reaction OH + D2 --> HOD + D. AB - Experiments, employing crossed molecular beams, with vibrational state resolution have been performed on the simplest four-atom reaction, OH + D2 --> HOD + D. In good agreement with the most recent quantum scattering predictions, mode-specific reaction dynamics is observed, with vibration in the newly formed oxygen deuterium bond preferentially excited to v = 2. This demonstrates that quantum theoretical calculations, which in the past decade have achieved remarkable accuracy for three-atom reactions involving three dimensions, have progressed to the point where it is now possible to accurately predict energy disposal in four atom reactions involving six dimensions. PMID- 11062123 TI - First-principles theory for the H + H2O, D2O reactions. AB - A full quantum dynamical study of the reactions of a hydrogen atom with water, on an accurate ab initio potential energy surface, is reported. The theoretical results are compared with available experimental data for the exchange and abstraction reactions in H + D2O and H + H2O. Clear agreement between theory and experiment is revealed for available thermal rate coefficients and the effects of vibrational excitation of the reactants. The excellent agreement between experiment and theory on integral cross sections for the exchange reaction is unprecedented beyond atom-diatom reactions. However, the experimental cross sections for abstraction are larger than the theoretical values by more than a factor of 10. Further experiments are required to resolve this. PMID- 11062124 TI - A light-emitting field-effect transistor. AB - We report here on the structure and operating characteristics of an ambipolar light-emitting field-effect transistor based on single crystals of the organic semiconductor alpha-sexithiophene. Electrons and holes are injected from the source and drain electrodes, respectively. Their concentrations are controlled by the applied gate and drain-source voltages. Excitons are generated, leading to radiative recombination. Moreover, above a remarkably low threshold current, coherent light is emitted through amplified spontaneous emission. Hence, this three-terminal device is the basis of a very promising architecture for electrically driven laser action in organic semiconductors. PMID- 11062125 TI - Emissions of methyl halides and methane from rice paddies. AB - Methyl halide gases are important sources of atmospheric inorganic halogen compounds, which in turn are central reactants in many stratospheric and tropospheric chemical processes. By observing emissions of methyl chloride, methyl bromide, and methyl iodide from flooded California rice fields, we estimate the impact of rice agriculture on the atmospheric budgets of these gases. Factors influencing methyl halide emissions are stage of rice growth, soil organic content, halide concentrations, and field-water management. Extrapolating our data implies that about 1 percent of atmospheric methyl bromide and 5 percent of methyl iodide arise from rice fields worldwide. Unplanted flooded fields emit as much methyl chloride as planted, flooded rice fields. PMID- 11062126 TI - Early Permian bipedal reptile. AB - A 290-million-year-old reptilian skeleton from the Lower Permian (Asselian) of Germany provides evidence of abilities for cursorial bipedal locomotion, employing a parasagittal digitigrade posture. The skeleton is of a small bolosaurid, Eudibamus cursoris, gen. et sp. nov. and confirms the widespread distribution of Bolosauridae across Laurasia during this early stage of amniote evolution. E. cursoris is the oldest known representative of Parareptilia, a major clade of reptiles. PMID- 11062127 TI - A kingdom-level phylogeny of eukaryotes based on combined protein data. AB - Current understanding of the higher order systematics of eukaryotes relies largely on analyses of the small ribosomal subunit RNA (SSU rRNA). Independent testing of these results is still limited. We have combined the sequences of four of the most broadly taxonomically sampled proteins available to create a roughly parallel data set to that of SSU rRNA. The resulting phylogenetic tree shows a number of striking differences from SSU rRNA phylogeny, including strong support for most major groups and several major supergroups. PMID- 11062128 TI - Recovery and management options for spring/summer chinook salmon in the Columbia River basin. AB - Construction of four dams on the lower Snake River (in northwestern United States) between 1961 and 1975 altered salmon spawning habitat, elevated smolt and adult migration mortality, and contributed to severe declines of Snake River salmon populations. By applying a matrix model to long-term population data, we found that (i) dam passage improvements have dramatically mitigated direct mortality associated with dams; (ii) even if main stem survival were elevated to 100%, Snake River spring/summer chinook salmon (Oncorhynchus tshawytscha) would probably continue to decline toward extinction; and (iii) modest reductions in first-year mortality or estuarine mortality would reverse current population declines. PMID- 11062129 TI - VirB/D4-dependent protein translocation from Agrobacterium into plant cells. AB - The Agrobacterium VirB/D4 transport system mediates the transfer of a nucleoprotein T complex into plant cells, leading to crown gall disease. In addition, several Virulence proteins must somehow be transported to fulfill a function in planta. Here, we used fusions between Cre recombinase and VirE2 or VirF to directly demonstrate protein translocation into plant cells. Transport of the proteins was monitored by a Cre-mediated in planta recombination event resulting in a selectable phenotype and depended on the VirB/D4 transport system but did not require transferred DNA. PMID- 11062130 TI - A basal transcription factor that activates or represses transcription. AB - We have identified an activity that is required for transcription of downstream promoter element (DPE)-containing core promoters in vitro. The purified factor was found to be the Drosophila homolog of the transcriptional repressor known as NC2 or Dr1-Drap1. Purified recombinant dNC2 activates DPE-driven promoters and represses TATA-driven promoters. A mutant version of dNC2 can activate DPE promoters but is unable to repress TATA promoters. Thus, the activation and repression functions are distinct. These studies reveal that NC2 (Dr1-Drap1) is a bifunctional basal transcription factor that differentially regulates gene transcription through DPE or TATA box motifs. PMID- 11062131 TI - Oxidative damage linked to neurodegeneration by selective alpha-synuclein nitration in synucleinopathy lesions. AB - Aggregated alpha-synuclein proteins form brain lesions that are hallmarks of neurodegenerative synucleinopathies, and oxidative stress has been implicated in the pathogenesis of some of these disorders. Using antibodies to specific nitrated tyrosine residues in alpha-synuclein, we demonstrate extensive and widespread accumulations of nitrated alpha-synuclein in the signature inclusions of Parkinson's disease, dementia with Lewy bodies, the Lewy body variant of Alzheimer's disease, and multiple system atrophy brains. We also show that nitrated alpha-synuclein is present in the major filamentous building blocks of these inclusions, as well as in the insoluble fractions of affected brain regions of synucleinopathies. The selective and specific nitration of alpha-synuclein in these disorders provides evidence to directly link oxidative and nitrative damage to the onset and progression of neurodegenerative synucleinopathies. PMID- 11062132 TI - Role of BAX in the apoptotic response to anticancer agents. AB - To assess the role of BAX in drug-induced apoptosis in human colorectal cancer cells, we generated cells that lack functional BAX genes. Such cells were partially resistant to the apoptotic effects of the chemotherapeutic agent 5 fluorouracil, but apoptosis was not abolished. In contrast, the absence of BAX completely abolished the apoptotic response to the chemopreventive agent sulindac and other nonsteroidal anti-inflammatory drugs (NSAIDs). NSAIDs inhibited the expression of the antiapoptotic protein Bcl-XL, resulting in an altered ratio of BAX to Bcl-XL and subsequent mitochondria-mediated cell death. These results establish an unambiguous role for BAX in apoptotic processes in human epithelial cancers and may have implications for cancer chemoprevention strategies. PMID- 11062133 TI - A PEST-like sequence in listeriolysin O essential for Listeria monocytogenes pathogenicity. AB - Establishment and maintenance of an intracellular niche are critical to the success of an intracellular pathogen. Here, the pore-forming protein listeriolysin O (LLO), secreted by Listeria monocytogenes, was shown to contain a PEST-like sequence (P, Pro; E, Glu; S, Ser; T, Thr) that is essential for the virulence and intracellular compartmentalization of this pathogen. Mutants lacking the PEST-like sequence entered the host cytosol but subsequently permeabilized and killed the host cell. LLO lacking the PEST-like sequence accumulated in the host-cell cytosol, suggesting that this sequence targets LLO for degradation. Transfer of the sequence to perfringolysin O transformed this toxic cytolysin into a nontoxic derivative that facilitated intracellular growth. PMID- 11062134 TI - Role of Bacillus subtilis SpoIIIE in DNA transport across the mother cell prespore division septum. AB - The SpoIIIE protein of Bacillus subtilis is required for chromosome segregation during spore formation. The COOH-terminal cytoplasmic part of SpoIIIE was shown to be a DNA-dependent adenosine triphosphatase (ATPase) capable of tracking along DNA in the presence of ATP, and the NH(2)-terminal part of the protein was found to mediate its localization to the division septum. Thus, during sporulation, SpoIIIE appears to act as a DNA pump that actively moves one of the replicated pair of chromosomes into the prespore. The presence of SpoIIIE homologs in a broad range of bacteria suggests that this mechanism for active transport of DNA may be widespread. PMID- 11062135 TI - Epithelial regulation of innate immunity to respiratory syncytial virus. PMID- 11062136 TI - Surfactant protein-A enhances uptake of respiratory syncytial virus by monocytes and U937 macrophages. AB - Surfactant protein (SP)-A is a known opsonin for a variety of pulmonary pathogens. SP-A enhances ingestion of these pathogens by interaction with an SP-A receptor (SP-AR) found on phagocytic cells such as peripheral blood monocytes (PBMC) and alveolar macrophages. Respiratory syncytial virus (RSV) is the most important respiratory pathogen in children. Recent studies have indicated that SP A levels may be decreased in RSV bronchiolitis and pneumonia. In this study we examined the role of SP-A in uptake of RSV by both PBMC and U937 macrophages, a human macrophage cell line known to express SP-ARs. In addition, we studied the effect of SP-A- mediated uptake of RSV on production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-10 by these cells because incomplete immunity to recurrent RSV infection has been partially attributed to abnormal cytokine responses by macrophages. SP-A enhanced binding and uptake of fluorescently labeled RSV (RSV-FITC) by PBMC in a dose-dependent manner, with a maximal effect seen with 10 to 15 microg/ml SP-A as measured by both percent fluorescent monocytes and linear mean fluorescence (lmf) of individual cells. SP-A also enhanced uptake of RSV-FITC by U937 macrophages, with a maximal effect seen with 20 microg/ml SP-A as measured by both percent fluorescent monocytes and lmf. With respect to TNF-alpha levels, RSV alone slightly enhanced TNF-alpha production by PBMC and decreased TNF-alpha production by U937 macrophages measured at 12 h after addition of RSV. SP-A-mediated uptake of RSV significantly enhanced TNF alpha production by PBMC and reversed the RSV-induced depression of TNF-alpha by U937 macrophages. RSV significantly enhanced IL-10 production by both cell types, which was reversed by SP-A-mediated uptake. These findings suggest that SP-A is an important opsonin for RSV and that SP-A-mediated uptake of RSV may alter some of the unusual cytokine responses that are postulated to be involved in incomplete immunity to recurrent infection. PMID- 11062137 TI - Hypoxia activates jun-N-terminal kinase, extracellular signal-regulated protein kinase, and p38 kinase in pulmonary arteries. AB - Chronic alveolar hypoxia is the major cause of pulmonary hypertension. The cellular mechanisms involved in hypoxia- induced pulmonary arterial remodeling are still poorly understood. Mitogen-activated protein kinase (MAPK) is a key enzyme in the signaling pathway leading to cellular growth and proliferation. The purpose of this investigation was to determine the roles that MAPKs, specifically Jun-N-terminal kinase (JNK), extracellular signal-regulated protein kinase (ERK), and p38 kinase, play in the hypoxia-induced pulmonary arterial remodeling. Rats were exposed to normobaric hypoxia (10% O(2)) for 1, 3, 7, or 14 d. Hypoxia caused significant remodeling in the pulmonary artery characterized by thickening of pulmonary arterial wall and increases in tissue mass and total RNA. JNK, ERK, and p38 kinase tyrosine phosphorylations and their activities were significantly increased by hypoxia. JNK activation peaked at Day 1 and ERK/p38 kinase activation peaked after 7 d of hypoxia. The results from immunohistochemistry show that hypoxia increased phospho-MAPK staining in both large and small intrapulmonary arteries. Hypoxia also upregulated vascular endothelial growth factor messenger RNA (mRNA) and platelet-derived growth factor receptor mRNA levels in pulmonary artery with a time course correlated to the activation of ERK and p38 kinase. The gene expressions of c-jun, c-fos, and egr-1, known as downstream effectors of MAPK, were also investigated. Hypoxia upregulated egr-1 mRNA but downregulated c-jun and c-fos mRNAs. These data suggest that hypoxia induced activation of JNK is an early response to hypoxic stress and that activation of ERK and p38 kinase appears to be associated with hypoxia-induced pulmonary arterial remodeling. PMID- 11062138 TI - Enhanced epithelial gene transfer by modulation of tight junctions with sodium caprate. AB - The airway epithelium is resistant to infection by gene transfer vectors when infected from the luminal surface. One strategy for enhancing airway epithelial gene transfer is to modify paracellular permeability, thereby permitting the diffusion of vectors to the basolateral surface, where uptake receptors are expressed. We investigated the ability of a medium-chain fatty acid known to enhance drug absorption, sodium caprate (C10), to increase airway paracellular permeability in comparison with ethyleneglycol-bis-(beta-aminoethyl ether)-N,N' tetraacetic acid (EGTA). Apical application of C10 decreased transepithelial resistance by > 90% within minutes, whereas EGTA required an hour or more to produce a similar effect. C10 increased mannitol and dextran permeability by sevenfold, as compared with a twofold increase produced by EGTA. A greater enhancement of adenoviral lacZ gene transfer was mediated by C10 (50-fold over controls) than by EGTA (10-fold over controls). This correlated with a significant enhancement of adenoviral CFTR-mediated correction of Cl(-) transport in polarized human airway epithelial (HAE) cells from cystic fibrosis (CF) patients. Confocal microscopy revealed a redistribution of claudin-1 following C10 but not EGTA treatment as a possible mechanism of gene-transfer enhancement by C10. These data suggest that C10 may be a better agent for enhancing gene transfer than is EGTA, and that this effect occurs through disruption of claudin 1. PMID- 11062139 TI - Loss of epithelial integrity resulting from E-cadherin dysfunction predisposes airway epithelial cells to adenoviral infection. AB - Epithelial intercellular adhesion is fundamental to the formation of the airway epithelial protective barrier. In this respect, cadherins are important because these adhesion molecules regulate formation and maintenance of epithelial intercellular junctions. To study the importance of airway epithelial integrity in determining susceptibility to virus infection, we used a replication incompetent adenovirus, RAd35, and an E-cadherin specific function-blocking antibody, SHE78-7, to disrupt intercellular contacts in human bronchial epithelial cell line 16HBE14o- and primary bronchial epithelial cells. After exposure of 16HBE14o- cell cultures to SHE78-7, disruption of the transepithelial permeability barrier was indicated by a loss of transepithelial electrical resistance and an associated increase of mannitol, inulin, and dextran paracellular flux. Subsequent exposure of SHE78-7-treated cell cultures to RAd35 showed a remarkable increase in adenoviral infection as assessed by beta galactosidase reporter gene expression. In cultures exposed to SHE78-7, disruption of E-cadherin function resulted in infection equivalent to that in control cultures using 16-fold lower viral titers. These studies show that manipulation of E-cadherin function provides a specific means of altering epithelial integrity that in turn determines resistance of airway epithelia to adenoviral infection. PMID- 11062140 TI - Contribution of upregulated airway endothelin-1 expression to airway smooth muscle and epithelial cell DNA synthesis after repeated allergen exposure of sensitized Brown-Norway rats. AB - Endothelin-1 is a potent bronchoconstrictor peptide with pro-inflammatory and growth-promoting properties. After exposure of sensitized Brown-Norway rats to six repeated ovalbumin exposures, there was an increase in pro-endothelin (ET)-1 messenger RNA compared with saline-exposed control rats 24 h after the final exposure (P < 0.01). ET-1 immunoreactivity was increased sixfold in the bronchial epithelium of the larger conducting airways in the repeated allergen-exposed rats (P < 0.001). After repeated allergen exposure, there were increased rates of DNA synthesis in the airway smooth muscle (ASM) cells (P < 0.001) and epithelial cells (P < 0. 001) compared with saline-exposed controls, as measured by bromodeoxyuridine incorporation. Treatment with a dual endothelin A and B (ET(A+B)) receptor antagonist caused a significant attenuation in both ASM (P < 0.001) and epithelial cell (P < 0.001) bromodeoxyuridine incorporation compared with the allergen-challenged and vehicle-treated group. The dual ET(A+B) antagonist attenuated eosinophil recruitment into the airways (P < 0. 05) but had no significant effect on increased bronchial reactivity to acetylcholine in allergen-exposed rats. Increased levels of ET-1 in the airways may contribute to inflammation and ASM and epithelial cell DNA synthesis after repeated allergen exposure. Such processes may underlie increased proliferation of resident cells leading to airway wall remodeling in asthmatics. PMID- 11062141 TI - Effect of adenovector-mediated gene transfer of keratinocyte growth factor on the proliferation of alveolar type II cells in vitro and in vivo. AB - Alveolar type II cell proliferation occurs after lung injury and is thought to minimize the subsequent fibrotic response. Keratinocyte growth factor (KGF) has been shown to be a potent growth factor for rat alveolar type II cells. In this study, we created a replication-deficient, recombinant human type 5 adenovirus vector expressing human KGF (Ad5-KGF) to produce alveolar type II cell hyperplasia in vivo. In rat type II cells in vitro, Ad5-KGF at a multiplicity of infection (MOI) of 2, 4, and 8 plaque-forming units (PFU)/cell increased thymidine incorporation 13.3-, 16.8-, and 20. 8-fold, respectively. The KGF concentration in the medium increased up to 26.0 +/- 1.0 ng/ ml. We then instilled 10(9) PFU of Ad5-KGF, Ad5-LacZ, or phosphate-buffered saline into Fischer 344 rats and analyzed the lungs 2, 3, 7, 14, 21, and 28 d later. Ad5-KGF produced extensive alveolar type II cell hyperplasia on Days 2, 3, and 7. Surfactant protein (SP)-A and SP-D in lavage and SP-D in serum increased more in the Ad5-KGF group than in the Ad5-LacZ and PBS groups on Days 2 and 3. KGF was readily detectable for up to 7 d in lavage fluid, although only a modest number of cells expressed KGF messenger RNA as detected by in situ hybridization. These data show that Ad5-KGF stimulates extensive alveolar type II cell proliferation in vivo. PMID- 11062142 TI - Phenotypically different cells with heterogeneous nuclear ribonucleoprotein A2/B1 overexpression show similar genetic alterations. AB - Immunocytochemical studies have revealed that overexpression of heterogeneous nuclear ribonucleoprotein (hnRNP) A2/ B1 in exfoliated epithelial cells is a potentially useful marker of early lung cancer. This study analyzed the correlation of hnRNP A2/B1 expression with molecular alterations in phenotypically different epithelial cells of paraffin-embedded pulmonary tissues. Sections from 20 human subjects were analyzed immunohistochemically for expression of hnRNP A2/B1. Normal-appearing, hyperplastic, and malignant epithelial cells with and without hnRNP A2/B1 expression (n = 78) were microdissected and assessed for microsatellite alterations (MA) and loss of heterozygosity (LOH) (n = 14 markers) as well as for clonality. Results showed that (1) hnRNP A2/B1 immunoreactive cells contained a significantly higher frequency of MA and LOH than did comparable cells that lacked detectable hnRNP A2/B1; (2) over 80% of MA and LOH seen in hnRNP A2/B1 immunoreactive normal appearing and hyperplastic cells persisted in malignant cells; (3) preliminary analysis of methylation status of the androgen receptor gene in non-neoplastic cells was suggestive of hnRNP A2/B1-expressing cells being of clonal origin; and (4) cells with cytoplasmic hnRNP A2/B1 immunoreactivity had a 3-fold higher frequency of MA and LOH than did cells with nuclear hnRNP A2/B1 immunoreactivity. These findings suggest that phenotypically different respiratory epithelial cells with hnRNP A2/B1 overexpression might be clonally derived, and that the subcellular localization of hnRNP A2/B1 might be an important factor associated with tumor progression. PMID- 11062143 TI - CD40L, but not CD40, is required for allergen-induced bronchial hyperresponsiveness in mice. AB - Asthma is characterized by immunoglobulin (Ig) E production, infiltration of the respiratory mucosa by eosinophils (EOSs) and mononuclear cells, and bronchial hyperresponsiveness (BHR). Interaction of CD40 on B cells and antigen presenting cells, with its ligand (CD40L) expressed transiently on activated T cells, is known to augment both T cell-driven inflammation and humoral immune responses, especially IgE production. Considering both the prominent role of inflammation in asthma and the association of the disease with IgE, we hypothesized that CD40 CD40L interactions would be important in pathogenesis. To test this hypothesis, we subjected wild-type (WT) mice and animals lacking either CD40 or CD40L to repeated inhalation of Aspergillus fumigatus (Af ) antigen. Af-treated WT mice displayed elevated IgE levels, bronchoalveolar lavage and pulmonary tissue eosinophilic inflammation, and BHR. IgE production was markedly suppressed in both the CD40 -/- and CD40L -/- strains. However, pulmonary inflammation did not appear to be inhibited by either of these mutations. Paradoxically, development of BHR was prevented by the lack of CD40L but not by the absence of CD40. We conclude that CD40/CD40L interactions, although critical in the induction of IgE responses to inhaled allergen, are not required for the induction of EOS predominant inflammation. CD40L, but not CD40, is necessary for the development of allergen-induced BHR. PMID- 11062144 TI - Role of alveolar macrophages in innate immunity in neonates: evidence for selective lipopolysaccharide binding protein production by rat neonatal alveolar macrophages. AB - As the first line of defense against inhaled substances, alveolar macrophages (AM) play a crucial role in maintaining lung homeostasis. This is achieved via phagocytosis of foreign material and the secretion of a wide range of mediator molecules, including those involved in neutrophil recruitment. Neonates are known to manifest increased susceptibility to lung infections, and we hypothesize that this may be due in part to a deficiency in the function of AM. We report here that although recruitment of neutrophils into the respiratory tract of newborn animals in response to Moraxalla catarrhalis exposure is greatly delayed and diminished, AM from newborn animals have greater phagocytic capacity when compared with those from adult animals. Additionally, newborn AM respond normally to lipopolysaccharide (LPS) via production of a variety of chemokines, including macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, monocyte chemotactic protein-1, gro/ cytokine-induced neutrophil chemoattractant, MIP-2, and tumor necrosis factor-alpha. We have also demonstrated an LPS inducible expression of messenger RNA for LPS binding protein (LBP) in neonatal AM that was not observed in AM from adult animals or in peritoneal macrophages. We speculate that local production of LBP by AM may be a significant factor in the neonatal immunologic response to infections, providing a compensatory mechanism for the deficiency in specific neonatal immunity during this period of development when the newborn is being exposed to a range of potentially pathogenic materials for the first time. PMID- 11062145 TI - Respiratory syncytial virus stimulation of vascular endothelial cell growth Factor/Vascular permeability factor. AB - We hypothesized that respiratory syncytial virus (RSV)-induced pathologies could be mediated, in part, by vascular active cytokines elaborated during virus infection. To address this hypothesis, we determined whether RSV stimulated vascular endothelial cell growth factor (VEGF)/vascular permeability factor (VPF) elaboration in vitro. Supernatants from unstimulated A549 cells and normal human bronchial epithelial cells contained modest levels of VEGF. In contrast, supernatants from RSV-infected cells contained elevated levels of VEGF/VPF. This stimulation was seen after as little as 2 h, was still prominent after 48 h, and, by immunoblot, was specific for the 165- and 121-amino acid isoforms of VEGF/VPF. It was not associated with significant cell cytotoxicity or alterations in VEGF messenger RNA. It did, however, require new protein biosynthesis. In accordance with these findings, the 165- and 121-amino acid isoforms of VEGF/VPF were also found in the nasal washings from patients with RSV infections. These studies demonstrate that RSV is a potent stimulator of VEGF/VPF elaboration and that, in vitro, this stimulation is mediated via a noncytotoxic translational and/or post translational biosynthetic mechanism. VEGF/VPF may play an important role in the pathogenesis of RSV-induced disorders. PMID- 11062146 TI - Zinc finger and carboxyl regions of adenovirus E1A 13S CR3 are important for transactivation of the cytomegalovirus major immediate early promoter by adenovirus. AB - Reactivation of latent cytomegalovirus (CMV) is an important cause of disease in susceptible patients. We previously demonstrated that an adenovirus early gene product can transactivate the CMV major immediate early (IE) promoter in inflammatory cells. This effect was due to the conserved region 3 (CR3) of the adenovirus E1A 13S gene product. There are two domains in the CR3 region, a zinc finger (aa 147-177) and a carboxyl (aa 180-188) domain. Both are crucial for transactivation of downstream promoter elements of adenovirus in E1A 13S. We sought to determine if either or both of these specific domains is also necessary for transactivation of the CMV IE promoter by the adenovirus E1A 13S gene product. We cotransfected T-lymphocyte Jurkat cells and monocyte/macrophage-like THP-1 cells with plasmids expressing wild-type (WT) or CR3 mutant E1A 13S and a CMV IE chloramphenicol acetyltransferase (CAT) reporter construct. With extracts of cells coinfected with E1A WT set to 100%, mutation in the zinc finger domain, the carboxyl domain, or both domains decreased CMV IE CAT activity by >/= 96%. In contrast, a mutation in the region between the zinc finger and carboxyl domains reduced CMV IE CAT activity by only 24 to 26%. Mixing studies in Jurkat cells confirmed the importance of these domains. We also evaluated the active site of the CMV IE promoter involved in transactivation in THP-1 cells using CMV IE promoter deletions and single promoter element constructs. These studies showed that progressive deletion of the 19-bp CMV IE repeats containing cyclic AMP response element binding protein/activating transcription factor (CREB/ATF) sites resulted in progressive loss of activity. The importance of this element was confirmed using single promoter elements containing CMV IE 16-, 18-, 19-, and 21 bp repeats. Finally, using a 19-bp single promoter element construct and the CR3 mutants we demonstrated that mutations in the zinc finger (C171S) carboxyl region (S185N) or both regions (C171S/ S185N) resulted in significant (83, 94, and 85%) loss of activity. We conclude that the zinc finger and carboxyl domains of the CR3 region of E1A 13S are necessary for transactivation of the CMV promoter and that this occurs mainly through activation of the 19-bp CREB/ATF site of the promoter. PMID- 11062147 TI - Polymorphism of human mucin genes in chest disease: possible significance of MUC2. AB - Most of the genes that encode epithelial mucins are highly polymorphic due to variations in the length of domains of tandemly repeated (TR) coding sequence, the part of the apomucin that is heavily glycosylated. We report here for the first time a difference in the distribution of MUC TR length alleles in chest disease. We examined the distribution of the length alleles of those MUC genes whose expression we have confirmed in the bronchial tree in an age- and sex matched series of 50 pairs of atopic patients with and without asthma. There was no significant difference in the distribution of alleles of MUC1, MUC4, MUC5AC, and MUC5B. MUC2, however, showed a highly significant difference in distribution. The atopic, nonasthmatic individuals showed an allele distribution that was very different from all our other patient and control groups, this group showing a longer mean allele length. The observations suggest that longer MUC2 alleles may help protect atopic individuals from developing asthma, though the effect may be due to a linked gene. The biological significance of this variation with respect to susceptibility to asthma will merit further investigation, and it will also be important to substantiate this finding on an independent data set. PMID- 11062148 TI - Epoxide formation from diallyl sulfone is associated with CYP2E1 inactivation in murine and human lungs. AB - We tested the hypothesis that an epoxide formed from diallyl sulfone (DASO(2)) is responsible for inactivation of CYP2E1 in murine and human lungs. An epoxide (1,2 epoxypropyl-3,3'-sulfonyl-1'-propene [DASO(3)]) was synthesized from DASO(2) and conjugated with glutathione (GSH) to produce the conjugates S-(1R, S-[[1 hydroxymethyl-2,3' -sulfonyl]-1' -propenyl]ethyl)glutathione (diastereomers) and S-(1-[[2R,S-hydroxypropyl]-3, 3'-sulfonyl]-1'-propenyl)glutathione (diastereomers). Analysis of these conjugates by high performance liquid chromatography revealed a major peak eluting at 20.5 min. This peak was detected in incubations of murine and human lung microsomes containing DASO(2) and nicotinamide adenine dinucleotide phosphate (NADPH), and was not detected in incubations performed in the absence of DASO(2) or NADPH. The amounts of epoxide derived GSH conjugates formed in the incubations were concentration-dependent and achieved saturation at 0.75 mM DASO(2). Formation of the conjugates was also time dependent and peaked at 2.0 h after DASO(2). The peak containing the GSH conjugates was also detected in incubations of CYP2E1-expressed lymphoblastoid microsomes, NADPH, and DASO(2). Maximal amounts of DASO(3), as estimated by formation of a 4-(p-nitrobenzyl)pyridine derivatized product, were detected in murine lung microsomes incubated for 35 min with 1 mM DASO(2). The derivatized DASO(3) was not detectable in incubations of human lung microsomes. p-Nitrophenol hydroxylation, a catalytic activity associated with CYP2E1, was reduced in murine and human lung microsomes incubated with DASO(2), with decreases that were concentration-dependent. Dose-dependent decreases in hydroxylase activity were also found in microsomes from mice treated in vivo with DASO(2) (25 to 200 mg/kg). These results supported the premise that an epoxide formed from DASO(2) mediates inactivation of lung CYP2E1. Furthermore, the findings suggested that the mouse model is relevant for studies of DASO(2) in human lung. PMID- 11062149 TI - Homozygosity mapping of a gene locus for primary ciliary dyskinesia on chromosome 5p and identification of the heavy dynein chain DNAH5 as a candidate gene. AB - Reduced mucociliary clearance in primary ciliary dyskinesia (PCD) causes recurrent infections of the upper and lower respiratory tract. The disease is usually inherited as an autosomal recessive trait. To identify a gene locus for PCD, we studied a large consanguineous family of Arabic origin. Direct examination of the respiratory cilia revealed ciliary akinesia. Electron microscopic examination of cilia showed absence of the outer dynein arms. Two of four affected individuals exhibited a situs inversus, typical for Kartagener syndrome, due to randomization of the left/right body axis. A total genome scan with 340 highly polymorphic microsatellites was performed. We localized a new gene locus for PCD to a region of homozygosity by descent on chromosome 5p15-p14 with a parametric multipoint logarithm of odds ratio (LOD) score of Zmax = 3.51 flanked by markers D5S2095 and D5S502 within an interval of 20 centimorgans sex averaged genetic distance. Applying a polymerase chain reaction-based approach, we identified a 1.5-kb partial complementary DNA of DNAH5 encoding a Chlamydomonas-related axonemal heavy dynein chain within the critical disease interval of this new PCD locus. On the basis of the Chlamydomonas model for PCD, this gene represents an excellent candidate for PCD. PMID- 11062150 TI - Esterification of all-trans-retinol in normal human epithelial cell strains and carcinoma lines from oral cavity, skin and breast: reduced expression of lecithin:retinol acyltransferase in carcinoma lines. AB - When exogenous [(3)H]retinol (vitamin A) was added to culture medium, normal human epithelial cells from the oral cavity, skin, lung and breast took up and esterified essentially all of the [(3)H]retinol within a few hours. As shown by [(3)H]retinol pulse-chase experiments, normal epithelial cells then slowly hydrolyzed the [(3)H]retinyl esters to [(3)H]retinol, some of which was then oxidized to [(3)H]retinoic acid (RA) over a period of several days. In contrast, cultured normal human fibroblasts and human umbilical vein endothelial cells (HUVEC) did not esterify significant amounts of [(3)H]retinol; this lack of [(3)H]retinol esterification was correlated with a lack of expression of lecithin:retinol acyltransferase (LRAT) transcripts in normal fibroblast and HUVEC strains. These results indicate that normal, differentiated cell types differ in their ability to esterify retinol. Human carcinoma cells (neoplastically transformed epithelial cells) of the oral cavity, skin and breast did not esterify much [(3)H]retinol and showed greatly reduced LRAT expression. Transcripts of the neutral, bile salt-independent retinyl ester hydrolase and the bile salt-dependent retinyl ester hydrolase were undetectable in all of the normal cell types, including the epithelial cells. These experiments suggest that retinoid-deficiency in the tumor cells could develop because of the lack of retinyl esters, a storage form of retinol. PMID- 11062151 TI - Expression of beta-catenin and full-length APC protein in normal and neoplastic colonic tissues. AB - Mutations of the APC gene are thought to be early events in the process of colorectal carcinogenesis. Although the complete function(s) of the APC gene product is not known, it has been shown that the APC protein interacts with beta catenin in a multi-protein complex to regulate the level of expression of beta catenin. Loss of normal APC protein function can lead to an accumulation of beta catenin in the cytosol and the nucleus. Immunohistochemical methods were used to determine the relationship between APC and beta-catenin protein expression in human colonic tissues (150 normal, 9 hyperplastic, 58 adenomas and 83 carcinomas) and 12 paired samples of normal and cancer tissue in mouse colon. In all samples of normal human and mouse colonic mucosa and in human hyperplastic polyps both APC and beta-catenin immunoreactivity were present in colonocytes. APC expression was cytoplasmic, with maximal immunoreactivity in the goblet cells. beta-Catenin expression was predominantly localized to the plasma membrane, with no nuclear immunoreactivity. APC immunoreactivity was absent in all of the mouse adenocarcinomas and 83% of the human colon cancers. All of the human and mouse carcinomas had nuclear and cytoplasmic beta-catenin expression. In contrast, only 29% of the 58 colonic adenomas were completely negative for APC immunoreactivity. Regardless of the presence or absence of APC, all of the adenomas had cytoplasmic and nuclear beta-catenin immunoreactivity. Many colonic adenomas retain expression of full-length APC protein whereas it is usually lost in colorectal cancers. Regardless of the status of APC protein expression, beta-catenin protein was found in the cytoplasm and nucleus of all neoplastic colonic mucosa. The dissociation between loss of expression of APC and accumulation of beta-catenin in the nucleus suggests that inactivation of both alleles of the APC gene may not be required for beta-catenin nuclear accumulation in colonic adenomas. PMID- 11062152 TI - Galectin-3 mediates genistein-induced G(2)/M arrest and inhibits apoptosis. AB - Many recent studies have focused on potential chemopreventive activities of dietary genistein, a natural isoflavonoid compound found in soy products. Genistein has been implicated in anticancer activities, including differentiation, apoptosis, inhibition of cell growth and inhibition of angiogenesis. In previous studies, genistein was shown to induce apoptosis and cell cycle arrest at G(2)/M in several cancer cell lines in vitro, which is associated with induction of p21(WAF1/CIP1), a universal inhibitor of cyclin dependent kinases. At present, the molecular basis for diverse genistein-mediated cellular responses is largely unknown. In the present study, we investigated whether galectin-3, an anti-apoptotic gene product, regulates genistein-mediated cellular responses. We show that genistein effectively induces apoptosis without detectable cell cycle arrest in BT549, a human breast epithelial cell line which does not express galectin-3 at a detectable level. In galectin-3 transfected BT549 cells, genistein induced cell cycle arrest at the G(2)/M phase without apoptosis induction. Interestingly, genistein induces p21(WAF1/CIP1) expression in galectin-3-expressing BT549 cells, but not in control BT549 cells undergoing apoptosis. Collectively, the results of the present study suggest that galectin 3, at least in part, is a critical determinant for genistein-mediated cell cycle arrest and apoptosis, and genistein induction of p21(WAF1/CIP1) is associated with cell cycle arrest, but not required for apoptosis induction. PMID- 11062153 TI - SULT1A1 catalyzes 2-methoxyestradiol sulfonation in MCF-7 breast cancer cells. AB - In a previous study of nine human breast-derived cell lines, rates of metabolism of 17beta-estradiol (E(2)) were greatly enhanced when cultures were exposed to the aromatic hydrocarbon receptor agonist, 2,3,7,8-tetrachlorodibenzo-p-dioxin. Elevated rates of E(2) hydroxylation at the C-2, -4, -6alpha and -15alpha positions were observed concomitant with the induction of cytochromes P450 1A1 and 1B1. In each cell line, 2- and 4-hydroxyestradiol (2- and 4-OHE(2)) were converted to 2- and 4-methoxyestradiol (2- and 4-MeOE(2)) by the action of catechol O:-methyltransferase. In this study, conjugation of these estrogen metabolites was investigated. A comparison of the levels of metabolites determined with and without prior treatment of the media with a crude beta glucuronidase/sulfatase preparation showed that most of the 2-MeOE(2) present was in conjugated form, whereas 4-MeOE(2), 6alpha-OHE(2) and 15alpha-OHE(2) were minimally conjugated. Inhibitor studies suggested that it was the sulfatase activity of the preparation that hydrolyzed the 2-MeOE(2) conjugates in MCF-7 cell media; the presence of 2-MeOE(2)-3-sulfate in MCF-7 culture media was confirmed by electrospray ion-trap mass spectrometry. To identify the enzyme catalyzing this conjugation, the expression of mRNAs encoding five sulfotransferases (SULT1A1, SULT1A2, SULT1A3, SULT1E1 and SULT2A1) was evaluated in the nine cell lines by use of the reverse transcription-polymerase chain reaction. Only expression of SULT1A1 mRNA correlated with the observed conjugation of nanomolar levels of 2-MeOE(2) in these cell lines. Cloning and sequencing of SULT1A1 cDNA from MCF-7 cells revealed that mRNAs encoding two previously identified allelic variants, SULT1A1*1 ((213)Arg) and SULT1A1*2 ((213)His), were expressed in these cells. Heterologous cDNA-directed expression of either variant in MDA-MB-231 cells, which do not normally express SULT1A1, conferred 2-MeOE(2) sulfonation activity. The SULT1A1 allelic variants were also expressed in SF:9 insect cells, from which post-microsomal supernatants were used to determine K:(m) values of 0.90 +/- 0.12 and 0.81 +/- 0.06 microM for SULT1A1*1 and SULT1A1*2, respectively, with 2-MeOE(2) as substrate. These results show that SULT1A1 is an efficient and selective catalyst of 2-MeOE(2) sulfonation and, as such, may be important in modulating the anticarcinogenic effects of 2-MeOE(2) that have been described recently. PMID- 11062154 TI - Effects of calorie restriction on thymocyte growth, death and maturation. AB - We previously reported that calorie restriction (CR) significantly delays the spontaneous development of thymic lymphomas and other neoplasms in p53-deficient mice and their wild-type littermates. The purpose of the present study was to further characterize the anti-lymphoma effects of CR by assessing thymocyte growth, death and maturation in response to acute (6 day) and chronic (28 day) CR regimens. Male C57BL/6J mice fed a CR diet (restricted to 60% of control ad libitum intake) for 6 days displayed a severe reduction in thymic size and cellularity, as well as a decrease in splenic size and cellularity; these declines were sustained through 28 days of CR. Mice maintained on a CR diet for 28 days also displayed a significant depletion in the cell numbers of all four major thymocyte subsets defined by CD4 and CD8 expression. Analysis within the immature CD4(-)8(-) thymocyte subset further revealed an alteration in normal CD44 and CD25 subset distribution. In particular, CR for 28 days resulted in a significant decrease in the percentage of the proliferative CD44(-)25(-) subset. In addition, a significant increase in the percentage of the early, pro-T cell CD44(+)25(-) population was detected, indicative of a CR-induced delay in thymocyte maturation. Taken together, these findings suggest that CR suppresses (through several putative mechanisms) lymphomagenesis by reducing the pool of immature thymocytes that constitute the lymphoma-susceptible subpopulation. PMID- 11062155 TI - Effects of black tea, green tea and wine extracts on intestinal carcinogenesis induced by azoxymethane in F344 rats. AB - We investigated whether polyphenolic extracts from black tea, green tea or red wine affect azoxymethane (AOM)-induced intestinal carcinogenesis. Male F344 rats were treated 10 times (1 week apart) with AOM (7.4 mg/kg, s.c.) and then allocated into groups receiving black tea, green tea or red wine extracts mixed in the diet at a dose of 50 mg/kg body weight for 16 weeks. In the rats treated with black tea or wine extracts, there were significantly fewer colorectal tumours than in controls (the mean +/- SE number of tumours/rat was 2.54 +/- 1.6 in controls, 1.54 +/- 1.4 in the black tea group, 3.2 +/- 1.9 in the green tea group and 1.63 +/- 1.6 in the wine extract group). Significantly fewer rats in the black tea and wine extract groups had adenomas than in controls (86%, 59%, 90% and 50% of rats in the control, black tea, green tea and wine extract groups, respectively, had adenomas). The tumours from the black tea group and, to a lesser extent, those from the wine group, had a significantly greater apoptotic index than tumours in controls (mean +/- SE apoptotic index: 2.92 +/- 0.25, 4.13 +/- 0.46, 2.88 +/- 0.30 and 3.72 +/- 0.46 in controls, black tea, green tea or wine extract groups, respectively). In contrast, the apoptotic index of the normal mucosa did not vary among groups. These data indicate that black tea and wine extracts, but not green tea extracts, can protect against AOM-induced colon carcinogenesis by a mechanism probably involving increased apoptosis in tumours. PMID- 11062156 TI - Glutathione S-transferase M1 polymorphism and lung cancer risk in African Americans. AB - Glutathione S:-transferase M1 (GSTM1) can detoxify many carcinogens, including polycyclic aromatic hydrocarbons such as those from cigarette smoke. Though a number of studies have been published about the association between GSTM1 polymorphism and lung cancer, this association has received limited attention in the African-American population. We conducted a case-control study to investigate the role of GSTM1 polymorphism in the development of lung cancer in African Americans. Specimens of DNA from 117 lung cancer cases and 120 controls were assayed for detection of GSTM1 genotype by polymerase chain reaction (PCR). The odds ratios (ORs) and 95% confidence intervals (CIs) for lung cancer associated with homozygous deletion of the GSTM1 gene and other risk factors were estimated by logistic regression. Thirty-seven of the 117 cases (31. 6%) and 24 of the 120 controls (20.0%) had the GSTM1 null genotype; the OR was 2.10 (95% CI 1.07-4.11) after adjustment for age, gender and smoking. The association was higher for squamous cell carcinoma (OR 2.98, 95% CI 1.09-8.19) than for adenocarcinoma (OR 1.95, 95% CI 0.81-4.66). We observed a stronger association between GSTM1 null genotype and lung cancer among heavy smokers with > or =30 pack-years (OR 4.35, 95% CI 1.16-16.23). This association was also found in squamous cell carcinoma (OR 6.26, 95% CI 1.31-29.91). In the analysis combining GSTM1 polymorphism and smoking, smokers with the null genotype had high risk (OR 8.19, 95% CI 2.35 28.62) compared with non-smokers with the wild-type genotype, and the risk increased with smoking cigarette pack-years (P: = 0.0001 for trend). Our results suggest that GSTM1 polymorphism plays a role in the development of lung cancer and modifies the risk for smoking-related lung cancer in African-Americans. PMID- 11062157 TI - Identification of single nucleotide polymorphisms in human DNA repair genes. AB - Variation in gene coding sequence represents a significant factor in predisposition to disease, including cancer. Variants of some DNA repair genes (e.g. MLH1, MSH2 and MSH6) are known to predispose to cancer. We identified single nucleotide polymorphisms (SNPs) in five DNA repair genes in 142 healthy individuals using a DNA sequencing protocol optimized for the direct detection of single nucleotide polymorphisms. This approach, called the heterozygote sequencing protocol (HSP), enables moderate-scale population surveys of SNPs. HSP uses fluorescently tagged primers and exploits the large dynamic range and low background of automated fluorescent sequencing. HSP may be used for any sequence that can be amplified by PCR. A total of 12 SNP variants in MGMT, ERCC1, CDK7, CCNH and XRCC4 were identified, 11 at polymorphic frequencies, with an average frequency of 0.22 (95% confidence interval 0.20-0.24). Among the 82 individuals for whom complete SNP profiles were available, no one person carried the GenBank reference sequence for all five genes. The extensive heterogeneity observed in these five genes is intriguing. All variants are in Hardy-Weinberg equilibrium, although the meaning of this equilibrium is unclear. Using this approach, possible associations of sequence variation, and hence of variation in DNA repair, with disease risk can be assessed. PMID- 11062158 TI - 7-Methylsulfinylheptyl and 8-methylsulfinyloctyl isothiocyanates from watercress are potent inducers of phase II enzymes. AB - Watercress is an exceptionally rich dietary source of beta-phenylethyl isothiocyanate (PEITC). This compound inhibits phase I enzymes, which are responsible for the activation of many carcinogens in animals, and induces phase II enzymes, which are associated with enhanced excretion of carcinogens. In this study, we show that watercress extracts are potent inducers of quinone reductase (QR) in murine hepatoma Hepa 1c1c7 cells, a widely adopted assay for measuring phase II enzyme induction. However, contrary to expectations, this induction was not associated with PEITC (which is rapidly lost to the atmosphere upon tissue disruption due to its volatility) or a naturally occurring PEITC-glutathione conjugate, but with 7-methylsulfinyheptyl and 8-methylsulfinyloctyl isothiocyanates (ITCs). While it was confirmed that PEITC does induce QR (5 microM required for a two-fold induction in QR), 7-methylsulfinyheptyl and 8 methylsulfinyloctyl ITCs were more potent inducers (0.2 microM and 0.5 microM, respectively, required for a two-fold induction in QR). Thus, while watercress contains three times more phenylethyl glucosinolate than methylsulfinylalkyl glucosinolates, ITCs derived from methylsulfinylalkyl glucosinolates may be more important phase II enzyme inducers than PEITC, having 10 - to 25-fold greater potency. Analysis of urine by liquid chromatography-mass spectroscopy (LC-MS) following consumption of watercress demonstrated the presence of N: acetylcysteine conjugates of 7-methylsulfinylheptyl, 8-methylsulfinyloctyl ITCs and PEITC, indicating that these ITCs are taken up by the gut and metabolized in the body. Watercress may have exceptionally good anticarcinogenic potential, as it combines a potent inhibitor of phase I enzymes (PEITC) with at least three inducers of phase II enzymes (PEITC, 7-methylsulfinylheptyl ITC and 8 methylsulfinyloctyl ITC). The study also demonstrates the application of LC-MS for the detection of complex glucosinolate-derived metabolites in plant extracts and urine. PMID- 11062159 TI - The role of nitric oxide in neoplastic transformation of C3H 10T1/2 embryonic fibroblasts. AB - Nitric oxide synthase inhibitors block the neoplastic transformation of C3H 10T1/2 cells in vitro. Evidence presented herein suggests that they mediate their effects early in the carcinogenic process as brief treatment with the NOS inhibitor aminoguanidine (AG) during log phase cell growth (initiation phase) is sufficient to block foci formation. In contrast, treatment initiated after formation of a confluent monolayer was associated with diminished protection, while treatment commencing late in the promotional phase had no protective effect and appeared to enhance the number and stage of foci observed. These findings suggest that while AG treatment can inhibit transformation during the early promotional phase, it most effectively inhibits transformation during the initiation phase. In general AG enhanced growth of both normal and tumor cells, suggesting that effects on growth were unrelated to its anti-transformation properties, however, these effects could be related to the effect on tumor cell stage noted above. Although induction of inducible nitric oxide synthase (iNOS) by treatment with LI during the last 2 weeks of the assay was associated with enhanced transformation, the efficacy of AG in protecting against transformation was not clearly associated with substantial reductions in NO synthesis. The data suggest that AG inhibits transformation early in the transformation process independently of iNOS inhibition and that AG may have deleterious effects late in the process, possibly through stimulation of tumor cell growth. PMID- 11062160 TI - Tumorigenicity of four optically active bay-region 3,4-diol 1, 2-epoxides and other derivatives of the nitrogen heterocycle dibenz[c,h]acridine on mouse skin and in newborn mice. AB - The nitrogen heterocycle dibenz[c,h]acridine (DB[c,h]ACR) and the enantiomers of the diastereomeric pair of bay-region 3,4-diol 1, 2-epoxides as well as other bay region epoxides and dihydrodiol derivatives of this hydrocarbon have been evaluated for tumorigenicity on mouse skin and in the newborn mouse. On mouse skin, a single topical application of 50 or 200 nmol of compound was followed 10 days later by twice-weekly applications of the tumor promoter 12-O: tetradecanoylphorbol-13-acetate for 20 weeks. DB[c, h]ACR and the four optically pure, bay-region 3,4-diol-1,2-epoxide isomers all had significant tumor- initiating activity. The isomer with (1R,2S,3S,4R) absolute configuration [(+)-DE 2] was the most active diol epoxide isomer. The (-)-(3R,4R)-dihydrodiol of DB[c, h]ACR, the expected metabolic precursor of the bay-region (+)-DE-2, was 4- to 6 fold more tumorigenic than its corresponding (+)-enantiomer. In tumorigenicity studies in newborn mice, a total dose of 70-175 nmol of DB[c,h]ACR or one of its derivatives was injected i.p. on days 1, 8 and 15 of life, and tumorigenic activity was determined when the mice were 36-39 weeks old. DB[c,h]ACR produced a significant number of pulmonary tumors and also produced hepatic tumors in male mice. Of the four optically active bay-region diol epoxides, only (+)-DE-2 and (+)-DE-1 with (1R,2S,3S,4R) and (1S, 2R,3S,4R) absolute configuration, respectively, produced a significant tumor incidence. At an equivalent dose, the (+)-DE-2 isomer produced several-fold more pulmonary tumors and hepatic tumors than the (+)-DE-1 isomer. The (-)-(3R,4R)-dihydrodiol, metabolic precursor of the bay-region (+)-DE-2, was strongly active and induced an equal number of pulmonary and hepatic tumors as did DB[c,h]ACR. The (+)-(3S,4S) dihydrodiol was less active. The bay-region (+)-(1R,2S)-epoxide of 1,2,3,4-tetrahydro DB[c,h]ACR was strongly tumorigenic in newborn mice whereas its (-)-(1S, 2R)-enantiomer was inactive. This contrasts with the data on mouse skin where both enantiomers had substantial tumorigenic activity. In summary, the bay-region (+)-(1R,2S,3S,4R) 3,4-diol 1,2-epoxide of DB[c,h]ACR was the most tumorigenic of the four optically active bay-region diol epoxides of DB[c,h]ACR on mouse skin and in the newborn mouse. These results with a nitrogen heterocycle are similar to earlier data indicating high tumorigenic activity for the R,S,S,R bay-region diol epoxides of several carbocyclic polycyclic aromatic hydrocarbons. PMID- 11062161 TI - Differentially expressed genes in asbestos-induced tumorigenic human bronchial epithelial cells: implication for mechanism. AB - Although exposure to asbestos fibers is associated with the development of lung cancer, the underlying mechanism(s) remains unclear. Using human papillomavirus immortalized human bronchial epithelial (BEP2D) cells, we previously showed that UICC chrysotiles can malignantly transform these cells in a stepwise fashion before they become tumorigenic in nude mice. In the present study we used cDNA expression arrays to screen differentially expressed genes among the tumorigenic cells. A total of 15 genes were identified, 11 of which were further confirmed by northern blot. Expression levels of these genes were then determined among transformed BEP2D cells at different stages of the neoplastic process, including non-tumorigenic cells that were resistant to serum-induced terminal differentiation, early and late passage transformed BEP2D cells, five representative tumor cell lines and fused tumorigenic-control cell lines which were no longer tumorigenic. A consistent 2- to 3-fold down-regulation of the DCC (deleted in colon cancer), Ku70 and heat shock protein 27 genes were detected in all the independently generated tumor cell lines while expression levels in early transformants as well as in the fusion cell lines remained normal. In contrast, all the tumor cell lines examined demonstrated 2- to 4-fold overexpression of the insulin receptor and its signal transduction genes. Differential expression of these genes was completely restored in the fusion cell lines examined. No alteration in c-jun or EGF receptor expression was found in any of the cell lines. Our data suggest that activation of the insulin receptor pathway and inactivation of DCC and Ku70 may cooperate in malignant transformation of BEP2D cells induced by asbestos. PMID- 11062162 TI - Quantitation of DNA and hemoglobin adducts and apurinic/apyrimidinic sites in tissues of F344 rats exposed to propylene oxide by inhalation. AB - Propylene oxide (PO) is a relatively weak mutagen that induces nasal tumor formation in rats during long-term inhalation studies at high exposures (> or =300 p.p.m.), concentrations that also cause cytotoxicity and increases in cell proliferation. Direct alkylation of DNA by PO leads mainly to the formation of N:7-(2-hydroxypropyl)guanine (7-HPG). In this study, the accumulation of 7-HPG in tissues of male F344 rats exposed to 500 p. p.m. PO (6 h/day, 5 days/week for 4 weeks) by the inhalation route was measured by gas chromatography-high resolution mass spectrometry (GC-HRMS). In animals killed up to 7 h following the end of the last exposure the levels of 7-HPG (pmol/micromol guanine) in nasal respiratory tissue, nasal olfactory tissue, lung, spleen, liver and testis DNA were 606.2 +/- 53.0, 297.5 +/- 56.5, 69.8 +/- 3.8, 43.0 +/- 3.8, 27.5 +/- 2.4 and 14.2 +/- 0.7, respectively. The amounts of 7-HPG in the same tissues of animals killed 3 days after cessation of exposure were 393.3 +/- 57.0, 222.7 +/- 29.5, 51.5 +/- 1.2, 26.7 +/- 1.0, 18.0 +/- 2.6 and 10.4 +/- 0.1. A comparable rate of disappearance of 7-HPG was found among all tissues. DNA from lymphocytes pooled from four rats killed at the end of the last exposure was found to have 39.6 pmol adduct/micromol guanine. Quantitation of DNA apurinic/apyrimidinic sites, potentially formed after adduct loss by chemical depurination or DNA repair, showed no difference between tissues from control and exposed rats. The level of N:-(2-hydroxypropyl)valine in hemoglobin of exposed rats was also determined using a modified Edman degradation method followed by GC-HRMS analysis. The value obtained was 90.2 +/- 10.3 pmol/mg globin. These data demonstrate that nasal respiratory tissue, which is the target tissue for carcinogenesis, has a much greater level of alkylation of DNA than non-target tissues. PMID- 11062163 TI - High frequency allelic loss on chromosome 17p13.3-p11.1 in esophageal squamous cell carcinomas from a high incidence area in northern China. AB - Allelic loss on chromosome 17p has been reported frequently in esophageal squamous cell carcinoma (ESCC) and generally encompasses the p53 locus at 17p13.1. However, a good correlation between allelic loss on 17p and mutation of p53 has not been found. This suggests the possibility that unknown tumor suppressor genes near p53 may be involved in the development of ESCC. To evaluate this possibility, we analyzed 30 microsatellite markers covering the entire short arm of chromosome 17 in 56 ESCC patients from a high risk population in northern China, including 34 with a family history of upper gastrointestinal (UGI) cancer and 22 without a family history of any cancer. Cancer lifestyle risk factors and clinical/pathological characteristics were also collected. We found frequent allelic loss (>/=65%) at 28 of the 30 markers evaluated in these ESCC patients. The highest frequencies of allelic loss (> or =80%) were found in three smaller regions: deletion region I located at 17p13.3-p13.2 (between D17S849 and D17S1828); deletion region II located at 17p13.2-p13.1 (between D13S938 and TP53); deletion region III located at 17p13.1-p12 (between D17S804 and D17S799). A number of genes have already been identified in these deleted regions, including: OVCA1, OVCA2 and HIC-1 in deletion region I; p53 in deletion region II; ZNF18, ZNF29, ALDH3 and ALDH10 in deletion region III. These results will help us direct future testing of candidate genes and narrow the search region for major new tumor suppressor genes that may play a role in the pathogenesis of ESCC. PMID- 11062164 TI - Cyclosporin A inhibits chromium(VI)-induced apoptosis and mitochondrial cytochrome c release and restores clonogenic survival in CHO cells. AB - A variety of key events in the molecular apoptotic pathway involve the mitochondria. Cyclosporin A (csA) affects the mitochondria by inhibiting the mitochondrial permeability transition (MPT), thereby preventing disruption of the transmembrane potential. The role of the MPT in apoptosis is not fully understood, but inhibition of the MPT may prevent the release of mitochondrial caspase activators, such as cytochrome c (cyt c), into the cytosol. Certain hexavalent chromium [Cr(VI)] compounds are known occupational respiratory tract toxins and carcinogens. We have previously shown that these compounds induce apoptosis as a predominant mode of cell death and that this effect can be observed in cell culture using soluble Cr(VI). We show here that Cr(VI)-induced apoptosis in Chinese hamster ovary (CHO) cells involves disruption of mitochondrial stability. Using a cyt c-specific monoclonal antibody, we observed a dose-dependent release of mitochondrial cyt c in cytosolic extracts of CHO cells exposed to apoptogenic doses of sodium chromate. Co-treatment of these cells with csA inhibited the release of cyt c and abrogated Cr(VI)-induced apoptosis as determined by a reduction in internucleosomal DNA fragmentation. Co treatment with csA also markedly increased clonogenic survival of Cr(VI)-treated CHO cells. In contrast, the general caspase inhibitor Z-VAD-FMK markedly inhibited most of the morphological and biochemical parameters of apoptosis but did not prevent cyt c release and did not increase clonogenic survival. These results suggest that the MPT plays an important role in the regulation of mitochondrial cyt c release and that this may be a critical point in the apoptotic pathway in which cells are irreversibly committed to death. PMID- 11062165 TI - Effect of black and green tea polyphenols on c-jun phosphorylation and H(2)O(2) production in transformed and non-transformed human bronchial cell lines: possible mechanisms of cell growth inhibition and apoptosis induction. AB - The biological activities of theaflavin (TF), theaflavin gallate (TFG) and theaflavin digallate (TFdiG) from black tea and (-)-epigallocatechin 3-gallate (EGCG) and (-)-epigallocatechin (EGC) from green tea were investigated using SV40 immortalized (33BES) and Ha-ras gene transformed (21BES) human bronchial epithelial cell lines. Growth inhibition and cell viability were measured by trypan blue dye exclusion assay following 24 h treatment with the tea polyphenols. TFdiG, EGC and EGCG displayed comparable inhibitory effects on the growth of 21BES cells, with estimated IC(50) values of 22-24 microM. TFG exhibited a lower inhibitory activity (IC(50) 37 microM) and TF was even less effective (IC(50) 47 microM) in this cell line. A similar effect was also observed in 33BES cells. These results suggest that the gallate structure of theaflavins is important for growth inhibition. Exposure of 21BES cells to 25 microM TFdiG, EGC and EGCG for 24 h led to induction of cell apoptosis/death as determined by the Annexin V apoptosis assay. With TFdiG treatment cell death occurred early, and quickly peaked at 8-12 h. Morphological observations showed that TFdiG-treated cells appeared irregular in shape, with cytoplasmic granules, suggesting a cytotoxic effect. On the other hand, EGC and EGCG showed a lag phase before a rapid increase in apoptosis between 16 and 24 h, without any marked morphological changes, which was similar to that induced by H(2)O(2). TFdiG, EGC and EGCG induced similar amounts of H(2)O(2) formation in 21BES cells. Exogenously added catalase significantly prevented EGC- and EGCG-induced cell apoptosis, but did not prevent TFdiG-induced cell death, suggesting that H(2)O(2) is involved in the apoptosis induced by EGCG and EGC, but not in TFdiG-induced cell death. EGCG and TFdiG were shown to decrease c-jun protein phosphorylation in 21BES cells. Such inhibition is expected to result in lowered AP-1 activity, which may contribute to the growth inhibitory activity of tea polyphenols. PMID- 11062166 TI - Potentiation of epidermal growth factor-induced DNA synthesis in rat hepatocytes by phenobarbitone: possible involvement of oxidative stress and kinase activation. AB - A transient induction of S phase DNA synthesis is a common feature of non genotoxic rodent hepatocarcinogens when administered in vivo. In the present study the ability of phenobarbitone (PB) to induce S phase DNA synthesis in primary cultures of rat hepatocytes was investigated. In the absence of serum or growth factors PB was not a mitogen per se. However, stimulation of S phase DNA synthesis by epidermal growth factor (EGF) was enhanced by co-culture with PB. This effect was both time and concentration dependent. The lowest concentration of PB that significantly enhanced the effect of EGF was 10 microM and the effect was maximal at 1.0 mM. At a concentration of 2.0 mM PB no longer enhanced EGF induced S phase DNA synthesis. Hepatocyte cultures pretreated with PB (0.1 mM) for 2 days were more responsive to the induction of S phase DNA synthesis by EGF for the subsequent 2 days. Despite the inhibition of PB enhancement of S phase DNA synthesis by the antioxidant dimethylthiourea, reduced glutathione was not depleted by PB treatment nor were oxidized glutathione or lipid peroxides elevated. Western blotting analysis showed that PB had no effect on epidermal growth factor receptor (EGFR) autophosphorylation per se after 1 and 48 h culture, enhanced sensitization of EGFR therefore does not appear to contribute to the enhancement of S phase DNA synthesis by PB. In contrast, treatment of hepatocytes with PB for 12 h resulted in a small but statistically significant activation of p42/44 MAP kinase activity and activation of protein kinase C, as measured by redistribution of enzyme activity from a soluble to a particulate compartment of hepatocytes. Therefore, PB-mediated changes in protein kinase activity may contribute to the potentiation this compound affords. PMID- 11062167 TI - Characterization of mutations induced by 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine in the colon of gpt delta transgenic mouse: novel G:C deletions beside runs of identical bases. AB - Mutations induced by one of the typical dietary mutagens/carcinogens, 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), were characterized using gpt delta transgenic mice. This transgenic mouse model has two selection methods to efficiently detect different types of mutations, i.e. 6-thioguanine selection for point mutations and Spi(-) selection for deletions. The mice were fed with a diet containing 400 p.p.m. PhIP for 13 weeks and gpt and Spi(-) mutations were analyzed from the colon, where the highest mutant frequencies were detected. Concerning the types of gpt mutations from PhIP-treated mice, 81% were single base pair substitutions and G:C-->T:A transversions predominated; single base pair deletions at G:C base pairs were also observed. In untreated mice G:C-->A:T transitions predominated and >80% of these events involved 5'-CpG-3' sites. Concerning Spi(-) mutants from PhIP-treated mice, 76% were G:C base pair deletions and more than half of these events occurred in monotonic G or C run sequences. Interestingly, a novel type of frameshift motif, i.e. G:C base pair deletions beside run sequences, was observed. The most frequently observed mutation in this class was the 5'-TTTTTTG-3'-->5'-TTTTTT-3' event. These results suggest that PhIP induces point mutations, such as base substitutions and single base pair deletions, rather than larger deletions in vivo and that run sequences may play an important role in PhIP-induced G:C base pair deletions. PMID- 11062168 TI - Correlations of partial and extensive methylation at the p14(ARF) locus with reduced mRNA expression in colorectal cancer cell lines and clinicopathological features in primary tumors. AB - p14(ARF) is a putative tumor suppressor gene thought to modify the levels of p53. CpG sites within the 5'-flanking region and exon 1beta of p14(ARF) are targets of aberrant methylation and transcriptional silencing in human colorectal cancer (CRC). Here we have developed methylation-specific polymerase chain reaction (MSPCR) methods to detect methylation of CpG sites in p14(ARF) in CRC cell lines and primary CRC tumors, and correlated p14(ARF) mRNA expression with methylation in CRC cell lines using competitive quantitative reverse transcription-polymerase chain reaction methods. Ten CRC cell lines were studied; three (DLD-1, HCT15 and SW48) showed extensive methylation and six (Colo320, SW480, HT29, Caco2, SW837 and WiDr) were unmethylated; the other cell line, LoVo, showed partial methylation that affected exon 1beta but not the immediate upstream CpG sites. p14(ARF) mRNA was expressed at extremely low levels in fully methylated cell lines and at 10(4)- to 10(5)-fold higher levels in unmethylated cell lines. p14(ARF) expression in the partially methylated LoVo cell line was intermediate. Treatment of LoVo cells with 2 microM 5-aza-2'-deoxycytidine for 72 h was associated with marked (100-fold) induction of mRNA levels. Of 119 primary CRCs, 18% contained p14(ARF) methylation, although partial methylation was the most common pattern observed (in 67% of methylated tumors). Methylation of p14(ARF) was often accompanied by p16(INK4a) methylation; however, 50% of p14(ARF) methylated tumors contained unmethylated p16(INK4a). Methylation at p14(ARF) was associated with female gender, greater age, proximal anatomic location and poor differentiation, but not stage at diagnosis. A two-step MSPCR method for assaying p14(ARF) methylation in human tumors is described. PMID- 11062169 TI - Identification of urine metabolites of 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine following consumption of a single cooked chicken meal in humans. AB - Many studies suggest that mutagenic/carcinogenic chemicals in the diet, like 2 amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), may play a role in human cancer initiation. We have developed a method to quantify PhIP metabolites in human urine and have applied it to samples from female volunteers who had eaten a meal of cooked chicken. For this analysis, urine samples (5 ml) were spiked with a deuterium-labeled internal standard, adsorbed to a macroporous polymeric column and then eluted with methanol. After a solvent exchange to 0.01 M HCl, the urine extracts were passed through a filter, applied to a benzenesulfonic acid column, washed with methanol/acid and eluted with ammonium acetate and concentrated on a C(18) column. The metabolites were eluted from the C(18) column and quantified by LC/MS/MS. In our studies of human PhIP metabolism, eight volunteers were fed 200 g of cooked chicken containing a total of 27 microg PhIP. Urine samples were collected for 24 h after the meal, in 6 h aliquots. Although no metabolites could be found in urine collected from volunteers before eating the chicken, four major human PhIP metabolites, N:(2)-OH-PhIP-N:(2)-glucuronide, PhIP-N:(2)-glucuronide, 4'-PhIP-sulfate and N:(2)-OH-PhIP-N:3-glucuronide, were found in the urine after the chicken meal. The volunteers in the study excreted 4-53% of the ingested PhIP dose in the urine. The rate of metabolite excretion varied among the subjects, however, in all of the subjects the majority of the metabolites were excreted in the first 12 h. Very little metabolite was detected in the urine after 18 h. In humans, N:(2)-OH-PhIP-N:(2) glucuronide is the most abundant urinary metabolite, followed by PhIP-N:(2)-glucuronide. The variation seen in the total amount, excretion time and metabolite ratios with our method suggests that individual digestion, metabolism and/or other components of the diet may influence the absorption and amounts of metabolic products produced from PhIP. PMID- 11062170 TI - Role of the beta isoform of 14-3-3 proteins in cellular proliferation and oncogenic transformation. AB - The 14-3-3 proteins are associated with proto-oncogene and oncogene products. Here, we generated NIH 3T3 cells overexpressing the beta isoform of the 14-3-3 proteins (14-3-3 beta) to examine the function of this isoform in cellular proliferation and oncogenic transformation. Overexpression of 14-3-3 beta in NIH 3T3 cells stimulated cell growth and supported anchorage-independent growth in soft agar medium and tumor formation in nude mice. To elucidate the molecular mechanisms of 14-3-3 beta-mediated NIH 3T3 transformation, we examined the activity of mitogen-activated protein kinase (MAPK) after serum stimulation. Overexpression of 14-3-3 beta augmented MAPK activity after serum stimulation, and MAPK activity correlated well with the amount of 14-3-3 beta expression. The colony-forming ability of NIH 3T3 cells overexpressing 14-3-3 beta in soft agar medium was efficiently abolished by exogenous expression of a dominant-negative mutant of MEK1 and 14-3-3 beta physically interacted with Raf-1 in these cells. These findings indicate that 14-3-3 beta has oncogenic potential, mainly through enhancement of Raf-1 activation and resultant augmentation of signaling in the MAPK cascade. PMID- 11062171 TI - Duodenogastric reflux and foregut carcinogenesis: analysis of duodenal juice in a rodent model of cancer. AB - The incidence of esophageal adenocarcinoma is increasing rapidly. In rats, surgically induced duodenoesophageal reflux is carcinogenic. One proposed mechanism of carcinogenesis is based on the reaction of physiological bile acids with nitrite to produce carcinogenic N:-nitroso amides. To test this hypothesis, duodenal juice was analyzed for endogenously formed N:-nitroso bile acids and its genotoxicity was determined. Esophagojejunostomy was performed on 15 Sprague Dawley rats to produce duodeno-esophageal reflux. At the time of surgery and 2 and 6 weeks later, duodenal contents were aspirated and analyzed immediately. High performance liquid chromatography coupled to tandem mass spectrometry was used to detect bile acids and their nitroso derivates. Genotoxicity was assessed using a micronucleus test. The characteristic pattern of bile acid derivatives, with taurocholic acid (TCA) and glycocholic acid (GCA) as the predominant conjugates, was detected in all samples. However, even selective reaction monitoring experiments failed to demonstrate the presence of any N:-nitroso-TCA or N:-nitroso-GCA. In addition, other nitroso derivatives could not be detected in any of the samples by neutral loss experiments monitoring the loss of nitric oxide (detection limit 0.1% of the concentration of TCA). All samples were cytotoxic, but neither the preoperative nor the postoperative samples were genotoxic. Duodenal juice was cytotoxic but not genotoxic. Tumorigenesis of esophageal adenocarcinoma in the rodent model could not be linked to a specific carcinogen, especially not to nitroso bile acids. Chronic inflammation is likely to be the mechanism of carcinogenesis by duodenogastric reflux. PMID- 11062172 TI - Protective effect of topically applied olive oil against photocarcinogenesis following UVB exposure of mice. AB - Reactive oxygen species have been shown to play a role in ultraviolet light (UV) induced skin carcinogenesis. Vitamin E and green tea polyphenols reduce experimental skin cancers in mice mainly because of their antioxidant properties. Since olive oil has also been reported to be a potent antioxidant, we examined its effect on UVB-induced skin carcinogenesis in hairless mice. Extra-virgin olive oil was applied topically before or after repeated exposure of mice to UVB. The onset of UVB-induced skin tumors was delayed in mice painted with olive oil compared with UVB control mice. However, with increasing numbers of UVB exposures, differences in the mean number of tumors between UVB control mice and mice pretreated with olive oil before UVB exposure (pre-UVB group) were lost. In contrast, mice that received olive oil after UVB exposure (post-UVB group) showed significantly lower numbers of tumors per mouse than those in the UVB control group throughout the experimental period. The mean number of tumors per mouse in the UVB control, pre-UVB and post-UVB groups was 7.33, 6.69 and 2.64, respectively, in the first experiment, and 8.53, 9.53 and 3.36 in the second experiment. Camellia oil was also applied, using the same experimental protocol, but did not have a suppressive effect. Immunohistochemical analysis of DNA damage in the form of cyclobutane pyrimidine dimers (CPD), (6-4) photoproducts and 8 hydroxy-2'-deoxyguanosine (8-OHdG) in samples taken 30 min after a single exposure of UVB showed no significant difference between UVB-irradiated control mice and the pre-UVB group. In the post-UVB group, there were lower levels of 8 OHdG in epidermal nuclei, but the formation of CPD and (6-4) photoproducts did not differ. Exposure of olive oil to UVB before application abrogated the protective effect on 8-OHdG formation. These results indicate that olive oil topically applied after UVB exposure can effectively reduce UVB-induced murine skin tumors, possibly via its antioxidant effects in reducing DNA damage by reactive oxygen species, and that the effective component may be labile to UVB. PMID- 11062173 TI - Influence of Helicobacter pylori on reactive oxygen-induced gastric epithelial cell injury. AB - Risk factors for gastric cancer are receiving renewed attention in light of the recent positive association of Helicobacter pylori infection with gastric cancer. The effect of H.pylori on the balance between oxidants and antioxidants in the stomach is not well known. In this study, we investigated if exposure of gastric cells to H. pylori increases oxidant-associated gastric epithelial cell injury. A human gastric epithelial cell line (AGS) was grown on 96-well clusters, then exposed overnight to either live H.pylori (four cagA(+) and four cagA(-)) or broth culture supernatant from an isogenic H.pylori cagA(+) strain with and without vacA activity. Incubation of AGS cells with cagA(+) and cagA(-) H.pylori strains before exposure to reactive oxygen species (ROS) reduced cell viability on average to 73.7% and 39.5% of controls, respectively. The percent viability of cells exposed to ROS after incubation with control broth, vacA(-) broth and vacA(+) broth was 97.7%, 70.5% and 63.5%, respectively. Experiments were then performed to evaluate the effects of H.pylori exposure on the activities of ROS scavenging enzymes [catalase, glutathione peroxidase and superoxide dismutase (SOD)] and formation of 8-hydroxy-2-deoxyguanosine (8-OH-dG) adducts in AGS cells. Overnight exposure to cagA(-) strains reduced catalase activity by 42%; in contrast, exposure to cagA(+) H.pylori strains increased catalase activity by 51%. Glutathione peroxidase activity increased with exposure to both cagA(-) and cagA(+) strains by 95% and 240%, respectively. Total SOD activity increased 156% after exposure to cagA(+) strains and was marginally increased (52%) with exposure to cagA(-) strains. CuZn-SOD protein levels, assayed by enzyme-linked immunosorbent assay, were not significantly altered by exposure to H.pylori strains; however, Mn-SOD concentrations were significantly increased (P: < 0.02) after exposure to both cagA(-) and cagA(+) H.pylori strains. Exposure of AGS cells to cagA(+) and cagA(-) H.pylori was associated with, on average, 44.5 and 99.0 8-OH-dG/10(6) dG, respectively. The increase in catalase, glutathione peroxidase and SOD activity is associated with fewer 8-OH-dG DNA adducts and reduced susceptibility of AGS cells to lethal injury from ROS after exposure to cagA(+) H.pylori strains when compared with exposure to cagA(-) H.pylori strains. Alteration in the activity of ROS-scavenging enzymes by the presence of H. pylori may in part be responsible for the increased risk of gastric cancer in persons infected with H.pylori. PMID- 11062174 TI - Differential effects of toxic metal compounds on the activities of Fpg and XPA, two zinc finger proteins involved in DNA repair. AB - Even though not mutagenic, compounds of the carcinogenic metals nickel, cadmium, cobalt and arsenic have been shown previously to inhibit nucleotide excision repair and base excision repair at low, non-cytotoxic concentrations. Since some toxic metals have high affinities for -SH groups, we used the bacterial formamidopyrimidine-DNA glycosylase (Fpg protein) and the mammalian XPA protein as models to investigate whether zinc finger structures in DNA repair enzymes are particularly sensitive to carcinogenic and/or toxic metal compounds. Concentrations of Cys) involving a sequence change from 5'-TCGT to 5'-TTGT. We have shown previously that CPD formation by UVB or sunlight is enhanced up to 10-fold at 5'-CCG and 5'-TCG sequences due to the presence of 5-methylcytosine bases. Sequence analysis showed that the CpG at codon 270 is methylated in mouse epidermis at a level of approximately 85%. Irradiation of mouse skin or mouse cells in culture produced the strongest CPD signal within exon 8 at the 5'-TCG sequence which is part of codon 270. Time course experiments showed that CPDs at this particular sequence persist longer than at several neighboring positions. The data suggest that formation of CPDs is responsible for induction of the major p53 mutational hotspot in UV-induced mouse skin tumors. PMID- 11062177 TI - Interindividual variation in CYP1A1 expression in breast tissue and the role of genetic polymorphism. AB - The cytochrome P4501A1 (CYP1A1) enzyme is regulated at the transcriptional level and its expression is influenced by genetic factors, polymorphisms in the structural and regulatory genes, and by environmental factors such as exposure to polycyclic aromatic hydrocarbons (PAHs). To investigate the role of CYP1A1 in breast cancer, we studied CYP1A1 expression in breast tissue, thereby taking all possible modifying factors into account. We measured CYP1A1 expression in 58 non tumor breast tissue specimens from both breast cancer patients (n = 26) and cancer-free individuals (n = 32) using a newly developed reverse transcription polymerase chain reaction assay. CYP1A1 expression varied between specimens approximately 400-fold and was independent of age. CYP1A1 expression was somewhat higher in tissue from breast cancer patients than in that from cancer-free individuals, but this difference was not statistically significant. Analysis for CYP1A1 genetic polymorphisms revealed eight variants, seven in the cancer-free group and one in the patient group. The variant genotype was not a good predictor of expression level. We conclude that high CYP1A1 expression could be a risk factor for breast cancer and that the known CYP1A1 polymorphisms are not good predictors of CYP1A1 expression. PMID- 11062178 TI - Ethanol and acetaldehyde enhance benzo[a]pyrene-DNA adduct formation in human mammary epithelial cells. PMID- 11062180 TI - Molecular cloning and characterization of the human KIN17 cDNA encoding a component of the UVC response that is conserved among metazoans: by P.Kannouche, P.Mauffrey, G.Pinon-lataillade, M.G. Mattei, A.Sarasin, L.Daya-grosjean and J.F.Angulo PMID- 11062179 TI - Vitamin E inhibits cell proliferation and the activation of extracellular signal regulated kinase during the promotion phase of lung tumorigenesis irrespective of antioxidative effect. AB - We have already reported that the activation of extracellular signal-regulated kinase (Erk) is critical in the stimulation of cell proliferation during the promotion stage of urethane-induced lung tumorigenesis in mice. Also, we have found that vitamin E suppresses lung tumorigenesis by inhibiting cell proliferation at the promotion stage. However, it is still unclear whether this inhibitory effect at the promotion stage is based on the antioxidative effect of vitamin E or not. In order to address this question, we examined the inhibitory effect of alpha-tocopheryloxybutyric acid (TSE), an ether derivative of vitamin E that cannot act as an antioxidant in vivo, on cell proliferation and the activation of Erk during promotion of lung tumorigenesis. On day 30 after urethane injection (750 mg/kg, i. p.) in A/J mice, TSE or vitamin E at 100 micromol/kg, p.o. was administered. Twenty-four hours after the final administration, the mice were killed to analyze cell proliferation and related parameters. The labeling index of proliferating cell nuclear antigen (a marker of cell proliferation) and ornithine decarboxylase activity (a marker of the promotion stage in lungs) were attenuated by treatment with TSE or vitamin E. TSE or vitamin E treatment also inhibited urethane-induced activation of Erk and suppressed the activation of other essential members of the Erk cascade (Ras, Raf and Mek). These results suggest that vitamin E inhibits cell proliferation and activation of the Erk cascade during promotion of urethane-induced lung tumorigenesis in mice, independent of its antioxidative effect. PMID- 11062181 TI - Hypermethylation of the p16(Ink4a) promoter in B6C3F1 mouse primary lung adenocarcinomas and mouse lung cell lines: by A.C.Patel, C.H. Anna, J.F.Foley, P.S.Stockton, F.L.Tyson, J.C.Barrett and T.R. devereux PMID- 11062182 TI - Antibiotic clinical trials revisited. PMID- 11062183 TI - Antibiotics for Salmonella meningitis in children. PMID- 11062184 TI - Prevention of perinatal HIV transmission during pregnancy. AB - Transmission of the human immunodeficiency virus (HIV) from mother to child can occur in utero, during labour or after delivery from breastfeeding. The majority of infants are infected during delivery. Maternal HIV-1 plasma viral load at delivery is the most important predictor of vertical transmission. For this reason, efforts to interrupt transmission have focused on the use of antiretroviral therapy. Zidovudine has been shown to reduce significantly vertical HIV transmission when used antepartum and intrapartum by the mother and postpartum by the newborn for 6 weeks. However, zidovudine monotherapy increases the risk of developing zidovudine resistance and may jeopardize the goal of durable viral suppression and allow HIV disease progression in the mother and transmission to the infant. Potent antiretroviral therapy is now recommended for all HIV-infected pregnant women using the same criteria for non-pregnant individuals. If possible, combination antiretroviral regimens should include the use of zidovudine but not at the expense of long-term viral suppression. The use of elective Caesarean section should probably be reserved for women who fail to achieve viral suppression at the time of delivery or if indicated for obstetrical reasons. The practice of breastfeeding has been shown to diminish the long-term efficacy of perinatal antiretroviral therapy. All HIV-infected mothers should avoid breastfeeding the newborn if possible. This review summarizes major prospective and retrospective antiretroviral treatment studies in HIV-infected pregnant women. Pharmacokinetic information as it relates to pregnancy and adverse event profiles of antiretroviral agents are also discussed. The impact of recent advances in the management of HIV infection in pregnancy is discussed with regard to their feasibility in resource-poor countries. PMID- 11062185 TI - Application of fluoroquinolone pharmacodynamics. AB - Pharmacodynamics provides a rational basis for optimizing dosing regimens by describing the relationship between drug, host and antimicrobial effect. The successful identification of meaningful pharmacodynamic outcome parameters can, therefore, greatly assist clinicians in making objective prescribing decisions rather than relying on static in vitro MIC data. While pharmacodynamic outcome parameters have been proposed for select antimicrobial agents, their clinical application remains to be defined fully. Quinolone antibiotics are generally considered to have concentration-dependent bactericidal activity and peak/MIC and AUC/MIC ratios have been identified as possible pharmacodynamic predictors of clinical and microbiological outcome as well as the development of bacterial resistance. Investigators have suggested that AUC/MIC ratios of 100-125 or peak/MIC ratios of >10 are required to predict clinical and microbiological success and to limit the development of bacterial resistance. These conclusions are derived primarily from studies of Gram-negative bacteria, and recent data suggest that these ratios may not be applicable for Streptococcus pneumoniae, where an AUC/MIC ratio of <40 appears to be a more accurate predictor. There is considerable variation in pharmacodynamic calculations and outcome parameters appear to be quinolone- and pathogen-specific. Additional prospective clinical research is needed to characterize quinolone pharmacodynamic parameters and answer unresolved questions regarding optimal pharmacodynamic outcome predictors for Gram-positive bacteria, anaerobes and atypical respiratory pathogens. PMID- 11062186 TI - In vitro inactivation of Chlamydia trachomatis and of a panel of DNA (HSV-2, CMV, adenovirus, BK virus) and RNA (RSV, enterovirus) viruses by the spermicide benzalkonium chloride. AB - Kinetics of inactivation by the detergent spermicide benzalkonium chloride (BZK) of Chlamydia trachomatis and of a panel of DNA viruses [herpes simplex virus hominis type 2 (HSV-2), cytomegalovirus (CMV), adenovirus (ADV) and BK virus (BKV)] and RNA [respiratory syncytial virus (RSV) and enterovirus (ENV)] were established in accordance with a standardized in vitro protocol. After a 5 min incubation, inactivation of >95% of HSV-2 and CMV was obtained at a concentration of 0.0025% (w/v) (25 Ig/L); concentrations as low as 0.0005%, 0.0050% and 0.0125%, induced a 3.0 log10 reduction in infectivity of HSV-2 and CMV, RSV and ADV, respectively. After a 60 min incubation, concentrations of 0.0125% and 0.050% provided a 3.0 log10 reduction in infectivity of ENV and BKV, respectively. These features indicate that sensitivity to BZK was very high (HSV 2 and CMV) or high (RSV) for enveloped viruses, intermediate (ADV) or low (ENV and BKV) for non-enveloped viruses. Furthermore, BZK had marked antichlamydial activity, showing >99% killing after only a 1 min incubation at a concentration of 0.00125%. BZK demonstrates potent in vitro activity against the majority of microorganisms causing sexually transmitted infectious diseases, including those acting as major genital cofactors of human immunodeficiency virus transmission. These attributes qualify BZK as a particularly attractive candidate for microbicide development. PMID- 11062187 TI - Aeromonas hydrophila AmpH and CepH beta-lactamases: derepressed expression in mutants of Escherichia coli lacking creB. AB - The class 1 cephalosporinase (CepH) and class 2d oxacillinase (AmpH) from an Aeromonas hydrophila clinical isolate, strain T429125, have been cloned and sequenced. Both enzymes are typical of their equivalents in other species of Aeromonas Both cloned beta-lactamase genes were expressed at a low level in a standard laboratory Escherichia coli strain, but when cloned into a cre deletion E. coli mutant, they were expressed at significantly higher levels. Specific disruption of the creB gene resulted in similar increased levels of beta lactamase expression, so it was concluded that CreB represses the transcription of ampH and cepH in a cre(+) E. coli strain. The expression of cepH was four times that of ampH in the deltacre mutant because of an additional factor encoded on the cloned T429125 chromosomal fragment containing cepH. This factor was able to trans-activate expression of co-resident ampH in the deltacre mutant such that expression of the two genes was approximately equal. The entire cepH-containing fragment was sequenced, but it contained no genes that were obviously related to any known class of DNA-binding protein. PMID- 11062188 TI - Integron-located VEB-1 extended-spectrum beta-lactamase gene in a Proteus mirabilis clinical isolate from Vietnam. AB - A clinical isolate of Proteus mirabilis Lil-1 was obtained from a Vietnamese patient hospitalized in Paris, France. This isolate was resistant to cephalosporins, and there was marked synergy between cephalosporins and clavulanic acid together with unusual synergy between cefoxitin and cefuroxime. PCR analysis revealed the presence of blaVEB-1, an integron-located gene coding for an extended-spectrum beta-lactamase (ESBL) identified previously in an Escherichia coli isolate MG-1 from Vietnam. Using class 1 integron primers and blaVEB-1 intragenic primers, the insert region of the blaVEB-1-containing integron along with flanking sequences were amplified from P. mirabilis Lil-1 whole-cell DNA. A novel class 1 integron, In55, was identified that contained, in addition to intI1, qacEDelta1, sul1 and Orf5 genes, an 8 kb variable region. This region was comparable in size to that found previously in E. coli MG-1, but different from those previously identified in two Pseudomonas aeruginosa isolates from Thailand. In55 was located on a 190 kb self-transferable plasmid, which was different in size and structure from that found in E. coli MG-1. The finding of blaVEB-1 on different plasmids and integrons in enterobacterial isolates underlines the interspecies spread of this novel ESBL gene. PMID- 11062189 TI - Influence of different medium components on the in vitro activity of the growth promoting antibiotic flavomycin against enterococci. AB - The growth-promoting antibiotic flavomycin (also called bambermycin, flavophospholipol and moenomycin) has a complex spectrum of activity against enterococci, with some species being naturally resistant and others susceptible. In this study, proteins added to Mueller-Hinton II medium had a strong deleterious effect on the activity of flavomycin, glucose had no effect and starch decreased the activity of flavomycin. The fatty substances Tween 80 and tributyrin increased the activity of flavomycin for several enterococcal species. Slight differences in the composition of the susceptibility test medium affected the MIC results obtained, indicating that strict standardization of the test medium is necessary. PMID- 11062190 TI - Subinhibitory concentrations of erythromycin reduce pneumococcal adherence to respiratory epithelial cells in vitro. AB - We have investigated the influence of subinhibitory concentrations of erythromycin on the interaction between Streptococcus pneumoniae and human respiratory epithelial cells. Confluent in vitro cell cultures were inoculated with erythromycin-resistant S. pneumoniae and incubated for 24 h. Erythromycin significantly reduced adherence of the pneumococci after 4 h and 24 h: 4.0% +/- 0.7% (mean +/- S.E.M. ) of the pneumococci adhered to the epithelial cells in medium with erythromycin, compared with 7.7% +/- 0.8% in medium without erythromycin (P: = 0.002) after 4 h, and the corresponding values after 24 h were 24.2% +/- 5.3% and 38.4% +/- 5.0%, respectively (P: = 0.038). Disruption of epithelial integrity by S. pneumoniae, measured as the decrease in transepithelial electrical resistance, was delayed in the presence of erythromycin. Neither addition of erythromycin to the culture medium nor infection of the cell cultures with pneumococci significantly affected secretion of interleukin-8 by the epithelial cells. Addition of erythromycin to a pneumococcal suspension in cell culture medium without respiratory epithelial cells almost completely prevented the release of pneumolysin. We conclude that erythromycin at subinhibitory concentrations reduces the adherence to and disruption of respiratory epithelial cells by S. pneumoniae, possibly by interfering with pneumolysin release. PMID- 11062191 TI - Comparative pharmacodynamics of moxifloxacin and levofloxacin in an in vitro dynamic model: prediction of the equivalent AUC/MIC breakpoints and equiefficient doses. AB - To demonstrate the impact of the different pharmacokinetics of moxifloxacin and levofloxacin on their antimicrobial effects (AMEs), killing and regrowth kinetics of two clinical isolates of Staphylococcus aureus and one each of Escherichia coli and Klebsiella pneumoniae were studied. With each organism, a series of monoexponential pharmacokinetic profiles of single doses of moxifloxacin (T:1/2 = 12.1 h) and levofloxacin (T:(1/2) = 6.8 h) were simulated. The respective eight fold ranges of the ratios of area under the concentration-time curve (AUC) to the MIC were 58-475 and 114-934. Species- and strain-independent linear relationships observed between the intensity of AME (I:(E)) and log AUC/MIC were not superimposed for moxifloxacin and levofloxacin (r(2) = 0.99 in both cases). The predicted AUC/MIC ratios for moxifloxacin and levofloxacin that might be equivalent to Schentag's AUC/MIC breakpoint for ciprofloxacin (125) were estimated at 80 and 130, respectively. The respective equivalent MIC breakpoints were 0.41 mg/L (for a 400 mg dose of moxifloxacin) and 0.35 mg/L (for a 500 mg dose of levofloxacin). Based on the I:(E)-log AUC/MIC relationships, equiefficient 24 h doses (D:(24)s) of moxifloxacin and levofloxacin were calculated for hypothetical strains of S. aureus, E. coli and K. pneumoniae with MICs equal to the respective MIC50s (weighted geometric means of reported values). To provide an 'acceptable' I:(E) = 200 (log cfu/mL)*h, the D:(24)s of moxifloxacin for all three organisms were much lower (150, 30 and 60 mg, respectively) than the clinically proposed 400 mg dose. Although the usual dose of levofloxacin (500 mg) would be in excess for E. coli and K. pneumoniae (D:(24) = 36 and 220 mg, respectively), it might be insufficient for S. aureus (the estimated D:(24) = 850 mg). Moreover, to provide the same effect as a 400 mg D:(24) of moxifloxacin against staphylococci, levofloxacin would have to be given in a 5000 mg D:(24), which is 10-fold higher than its clinically accepted dose. The described method of generalization of data obtained with specific organisms to other representatives of the same species might be useful to predict the AMEs of new quinolones. PMID- 11062192 TI - A combined in vivo pharmacokinetic-in vitro pharmacodynamic approach to simulate target site pharmacodynamics of antibiotics in humans. AB - We describe a new approach to quantify in vivo anti-infective activity by simulating effect site pharmacokinetics of antibiotics in vitro. This approach is based on (i) the in vivo measurement of interstitial drug pharmacokinetics (PK) at the target site and (ii) a subsequent pharmacodynamic (PD) simulation of the time versus drug concentration profile in an in vitro setting. To demonstrate the feasibility of this approach, individual time-concentration profiles of ciprofloxacin were measured in the interstitial space fluid of eight healthy volunteers by microdialysis following iv administration of 200 mg. Thereafter, different isolates of Pseudomonas aeruginosa were exposed in vitro to the interstitial ciprofloxacin concentration profile obtained from in vivo experiments. This led to a 1- to 3-log10 decrease in the number of viable organisms after 8 h. Significant correlations were observed between the maximal bactericidal effect and several PK surrogate parameters, notably the AUC/MIC ratio (P: = 0.0005), the C:max/MIC ratio (P: = 0.006) and the time > MIC (P: = 0.02). Furthermore, the data were analysed with an integrated PK-PD model allowing a much more detailed evaluation of the data than using MIC. The model employed an E:max relationship to link unbound ciprofloxacin concentration to bacterial kill rate. In conclusion, our experiments show that therapeutic success and failure in antimicrobial therapy may be explained by pharmacokinetic variability at the target site. Therefore, the in vivo PK-in vitro PD approach presented in our study may provide valuable guidance for drug and dose selection of antimicrobial agents. PMID- 11062193 TI - Pharmacokinetics and comparative effects of telithromycin (HMR 3647) and clarithromycin on the oropharyngeal and intestinal microflora. AB - The pharmacokinetics in plasma and saliva of a new ketolide, telithromycin (HMR 3647), and the effect on the normal oropharyngeal and intestinal microflora were studied in healthy volunteers and compared with those of clarithromycin. Ten subjects received 800 mg telithromycin perorally once daily and 10 other subjects received 500 mg clarithromycin bid for 10 days. Blood, saliva and faecal specimens were collected at defined intervals before, during and after administration for pharmacokinetic and microbiological analyses. In subjects receiving telithromycin, the mean C:(max), AUC and C:(24) (24 h) in saliva exceeded the values obtained from plasma, while saliva and serum pharmacokinetic parameters were in the same range for the clarithromycin group. The quantitative ecological disturbances in the normal microflora during administration of telithromycin were moderate and comparable to those associated with clarithromycin administration. No overgrowth of yeasts or Clostridium difficile occurred. Emergence of resistant strains was seen in both treatment groups. Administration of both telithromycin and clarithromycin was associated with significant increases in MICs for intestinal Bacteroides isolates, which persisted 2 weeks after discontinuation of treatment. In addition, a significant emergence of highly clarithromycin-resistant alpha-haemolytic streptococci, intestinal enterococci and Enterobacteriaceae was detected at day 10 in the clarithromycin group. In conclusion, administration of telithromycin resulted in high drug levels in saliva, which indicates a good therapeutic profile for throat infections. Telithromycin seems to have a more favourable ecological profile compared with clarithromycin in terms of resistance development in the normal microflora. PMID- 11062194 TI - Efficacy of polycationic peptides in preventing vascular graft infection due to Staphylococcus epidermidis. AB - A rat model was used to investigate the efficacy of two polycationic peptides, ranalexin and buforin II, in the prevention of vascular prosthetic graft infection due to methicillin-resistant Staphylococcus epidermidis with intermediate resistance to glycopeptides. The in vitro activity of the peptides was compared with those of vancomycin and teicoplanin by MIC determination and time-kill study. Moreover, the efficacy of collagen-sealed peptide-soaked Dacron was evaluated in a rat model of graft infection. Graft infections were established in the dorsal subcutaneous tissue of 120 adult male Wistar rats. The in vivo study included a control group, one contaminated group that did not receive any antibiotic prophylaxis and four contaminated groups that received an antibiotic-soaked graft. Experiments demonstrated that the activities of buforin II and ranalexin were greater than those of vancomycin and teicoplanin. Particularly, rats with buforin II-coated Dacron grafts showed no evidence of staphylococcal infection while, for the rats with ranalexin-, vancomycin- and teicoplanin-coated Dacron grafts, the quantitative graft cultures demonstrated bacterial growth (1.9 x 10(2) +/- 0.6 x 10(2) cfu/mL, 6. 2 x 103 +/- 1.9 x 10(3) cfu/mL and 5.1 x 10(4) +/- 4.8 x 10(3) cfu/mL, respectively). The study demonstrated that the use of peptide-soaked Dacron graft can result in significant bacterial growth inhibition and indicates that these compounds may be potentially useful in prosthetic surgery. PMID- 11062195 TI - Molecular epidemiology of penicillin-resistant Streptococcus pneumoniae colonizing children with community-acquired pneumonia and children attending day care centres in Fortaleza, Brazil. AB - To study clonal diversity of penicillin-resistant Streptococcus pneumoniae, 161 randomly selected isolates with reduced susceptibility to penicillin, collected from the nasopharynx of children under 5 years of age with community-acquired pneumonia and healthy controls from public day-care and immunization centres in Fortaleza, Brazil, were characterized by microbiological and serological techniques and automated ribotyping. Also included were 44 randomly selected penicillin-susceptible strains and three international reference strains. With automated ribotyping 75 ribopatterns were observed: 50 ribogroups were unique and 25 ribogroups were represented by two or more isolates. Genetic diversity was extensive but some degree of genetic homogeneity was found in strains from children with pneumonia, strains from children in day-care centres, isolates with reduced susceptibility to penicillin and isolates expressing 'paediatric' serogroups. Fourteen (56%) clusters contained both isolates with reduced penicillin susceptibility and penicillin-susceptible isolates, suggesting emergence of penicillin resistance. In general, there was a good correlation between ribogroups and serogroups, but 12 (48%) clusters contained isolates with alternative serogroups. Isolates with such alternative serogroups were more often encountered in penicillin-susceptible strains (41%) than in strains with reduced susceptibility to penicillin (7%). Thirty-eight (19%) isolates (including seven penicillin-susceptible strains) showed ribotypes indistinguishable from those of two international epidemic clones of S. pneumoniae: ribogroup 54-S-1 (15 isolates) with a ribopattern characteristic of the 23F multiresistant 'Spanish/USA' clone and ribogroup 74-S-3 (23 isolates) with a pattern similar to that of the 6B multiresistant 'Spanish' clone. PMID- 11062196 TI - Streptococcus pneumoniae resistance to erythromycin and penicillin in relation to macrolide and beta-lactam consumption in Spain (1979-1997). AB - Streptococcus pneumoniae resistance to penicillin and erythromycin in relation to beta-lactam and macrolide consumption in Spain over 19 years (1979-1997) was studied from resistance data collected by a search of the literature. Antibiotic consumption was expressed in defined daily dosage (DDD)/1000 inhabitants/day. A significant relationship (P: < 0.001) between erythromycin resistance (MIC >/= 1 mg/L) and global macrolide consumption (r = 0.942), as well as between high-level penicillin resistance (MIC >/= 2 mg/L) and global beta-lactam consumption (r = 0.948) was observed. The relationship between erythromycin resistance and macrolide consumption was due mainly to consumption of macrolides taken twice a day (adjusted r(2) = 0.886). Prevalence of high-level penicillin resistance correlated with consumption of oral cephalosporins (adjusted r(2) = 0.877); however, there appeared to be no correlation of consumption of oral or parenteral aminopenicillins, narrow-spectrum penicillins or cephalosporins with intermediate level penicillin resistance (MIC 0. 12-1 mg/L). The prevalence of high-level penicillin and of erythromycin resistance were also strongly correlated with each other (r = 0.903, P: < 0.001). In addition to global consumption, different categories of resistance (high or intermediate), and the differential capability of antibiotics to select resistance, must be taken into account when studying antibiotic impact on bacterial populations. Although this ecological analysis is not able to demonstrate a causal relationship between antibiotic consumption and development of resistance, it suggests that overuse of certain specific antibiotics is more likely to be related to the increase in drug-resistant strains of S. pneumoniae. PMID- 11062197 TI - Treatment of methicillin-resistant staphylococcus aureus infections with quinupristin-dalfopristin in patients intolerant of or failing prior therapy. For the Synercid Emergency-Use Study Group. AB - Safety and efficacy of quinupristin-dalfopristin (an injectable streptogramin antibiotic) were evaluated in the treatment of a variety of infections due to methicillin-resistant Staphylococcus aureus (MRSA) in patients either intolerant of or failing prior therapy. The influence of resistance phenotypes on treatment outcome was also assessed. This worldwide, multicentre, open-label, non comparative, emergency-use clinical study enrolled patients with one or more of nine predefined, culture-confirmed infections with MRSA, who had no clinically appropriate alternative antibiotic therapy. The recommended quinupristin dalfopristin dose was 7.5 mg/kg administered iv every 8 h for a duration judged appropriate by the investigator. There were no restrictions on prior or concomitant treatment with other antibiotics. Clinical, microbiological and laboratory assessments were performed at baseline, during study drug treatment, within 24 h after the last dose, and 7-21 days post-therapy. Ninety patients [age (mean +/- S.D.) 57.4 +/- 18.5 years] with significant underlying medical illnesses were treated at 63 centres in five countries. The most common indications were bone and joint infection (44.4% of patients) and skin and skin structure infection (16.7%). The mean (+/- S.D.) daily dose and treatment duration was 20.2 +/- 2.9 mg/kg/day for 28.5 +/- 22.3 days, most frequently administered every 8 h. The overall success rate (defined as a clinical outcome of either cure or improvement and a bacteriological outcome of eradication or presumed eradication) was 71.1% in the all-treated population (n = 90) and 66.7% in patients who were both clinically and bacteriologically evaluable (n = 27). Success rates for endocarditis, respiratory tract infection and bacteraemia of unknown source were below the population mean. The macrolide-lincosamide streptogramin type B resistance phenotype did not appear to alter the response rate. The most common non-venous adverse events related to study medication were arthralgias (10.8%), myalgias (8.6%) and nausea (8.6%). Quinupristin-dalfopristin should be considered as a treatment option for infections caused by MRSA, especially in patients intolerant of or failing alternate therapy. PMID- 11062198 TI - Molecular analysis of the translational attenuator of a constitutively expressed erm(A) gene from Staphylococcus intermedius. AB - During the course of a study on macrolide, lincosamide and streptogramin B resistance among staphylococci from animal sources, a Staphylococcus intermedius isolate was found to carry a constitutively expressed erm(A) gene on the 70 kb plasmid pSES29. The molecular basis of constitutive erm(A) expression was investigated by cloning and sequence analysis of the erm(A)-associated translational attenuator. Two point mutations in this regulatory region were detected. These mutations cause constitutive erm(A) gene expression by destabilization of mRNA secondary structures required for the inducible type of erm(A) gene expression. PMID- 11062199 TI - Erythromycin resistance genes in group A streptococci of different geographical origins. The Macrolide Resistance Study Group. AB - A total of 238 erythromycin-resistant Streptococcus pyogenes isolates were collected in 1986-1997 from eight different countries in Europe and North and South America. The antibiotic susceptibility patterns of all isolates and the resistance genes of 92 isolates of known clonal origin were studied. The mefA gene was detected in all 54 isolates with the M-phenotype and was found in every country. The ermTR and the ermB genes were detected in 27 and 11, respectively, of the 38 isolates with the macrolide-lincosamide-streptogramin B resistance phenotypes. In addition to the mefA gene, the recently sequenced ermTR gene was also widely distributed among isolates of different clonal origin. PMID- 11062200 TI - Frameshift mutations in rdxA and metronidazole resistance in North American Helicobacter pylori isolates. AB - In Helicobacter pylori, the oxygen-insensitive nitroreductase RdxA is likely to activate metronidazole (Mtz) by reduction and formation of cytotoxic intermediates. Mutations in rdxA have been associated with Mtz resistance in H. pylori. In vitro Mtz susceptibilities of 17 randomly selected H. pylori isolates were determined by the agar dilution method. DNA sequence analysis of rdxA alleles of eight susceptible isolates (MIC range: 0.25-1.0 mg/L) and nine resistant isolates (MIC range: 16-256 mg/L) showed that six of nine Mtz-resistant H. pylori isolates contained insertion or deletion mutations ('indel' mutations). One isolate contained a substitution mutation at codon position 148 that resulted in the introduction of a premature stop codon. Creation of stop codons within the rdxA coding sequence by either frameshift or substitution mutations resulted in premature translation termination and expression of putatively truncated RdxA polypeptides. PMID- 11062201 TI - Comparative in vitro activity of telithromycin (HMR 3647), three macrolides, amoxycillin, cefdinir and levofloxacin against gram-positive clinical isolates in Japan. AB - The in vitro activities (MIC and MBC) of telithromycin (HMR 3647) against clinical isolates in Japan were investigated in comparison with those of erythromycin A, clarithromycin, azithromycin, amoxycillin, cefdinir and levofloxacin. Telithromycin was more potent than the reference compounds against erythromycin A-susceptible or resistant Streptococcus pneumoniae isolates possessing either mef (mefA or mefE) genes or the ermB gene. Against erythromycin A-susceptible or inducibly resistant Staphylococcus aureus and erythromycin A susceptible and intermediate Enterococcus faecalis, telithromycin was highly active. PMID- 11062202 TI - In vitro susceptibility of Helicobacter pylori to, and in vivo suppression by, antimicrobials used in selective decontamination of the digestive tract. AB - The incidence of bleeding related to stress ulcers is reduced in critically ill patients in whom gut decontamination has been performed; this may be a result of suppression of Helicobacter pylori infection. We determined the susceptibility of H. pylori to the applied antibiotics. In nine of 10 critically ill patients (using intravenous cefotaxime and topical polymyxin, tobramycin and amphotericin B (PTA) given by nasogastric tube) and all six volunteers (using PTA alone), H. pylori was suppressed as long as the topical antibiotics were ingested. The in vitro studies revealed that all strains were susceptible to cefotaxime and tobramycin. The strains were not susceptible to polymyxin or amphotericin B. PMID- 11062203 TI - Comparative activities of polycationic peptides and clinically used antimicrobial agents against multidrug-resistant nosocomial isolates of Acinetobacter baumannii. AB - The in vitro activity of buforin II, cecropin P1, indolicidin, magainin II and ranalexin, alone and in combination with eight clinically used antimicrobial agents was investigated against 12 multidrug-resistant strains of Acinetobacter baumannii isolated from immunocompromised patients. Antimicrobial activities were measured by MIC, MBC and viable count. The peptides had a varied range of inhibitory values: overall, the organisms were more susceptible to buforin II (MIC range 0.25-16 mg/L), cecropin P1 (0.50-32 mg/L) and magainin II (0.50-16 mg/L). Synergy occurred when magainin II was combined with beta-lactam antibiotics. PMID- 11062204 TI - Bactericidal effect of antibiotics on Bartonella and Brucella spp.: clinical implications. AB - The species Bartonella and Brucella are phylogenetically closely related bacteria, both of which can produce chronic infections in humans that are difficult to cure with antibiotics. MICs of antibiotics for both species correlate poorly with the in vivo efficacy of the antibiotics. In this study we have determined MBCs of several antibiotics for this group of pathogens. Only the aminoglycosides were bactericidal, and this correlates well with the usefulness of these antibiotics for the therapy of human brucellosis and chronic Bartonella spp. infections such as endocarditis. Our data indicate that current clinical experience in treating brucellosis may help to define better the optimum antibiotic therapy for Bartonella-related diseases. PMID- 11062205 TI - Susceptibility of Cryptococcus neoformans by the NCCLS microdilution and Etest methods using five defined media. AB - The susceptibility of 12 isolates of Cryptococcus neoformans to amphotericin B, 5 fluorocytosine, fluconazole, itraconazole and ketoconazole was tested using the NCCLS and Etest methods with yeast nitrogen base (YNB) pH 5.6 and pH 7.0, RPMI MOPS pH 7.0 with and without added glucose (2%) and RPMI buffered with phosphate buffer to pH 7.0. Some isolates yielded poor growth in RPMI MOPS after 72 h. Tests indicated that YNB pH 5.6 was the best medium for 5-fluorocytosine but was unsuitable for ketoconazole. In conclusion, YNB pH 7.0 or RPMI MOPS with 2% glucose can be used with either method. PMID- 11062206 TI - Metronidazole and albendazole susceptibility of 11 clinical isolates of Giardia duodenalis from France. AB - The metronidazole and albendazole susceptibility of 11 clinical isolates of Giardia duodenalis from France was determined using a neonatal mouse model and compared with the outcome in patients after standard metronidazole therapy (0.75 g/day for 5 days). All isolates found to be clinically resistant to metronidazole (4/11) exhibited an ID50 > 120 mg/kg in the mouse model. This therefore appears to be a suitable animal model in which to explore drug failures in human giardiasis. PMID- 11062207 TI - Impact of a new quinolone, DU-6859a, and two oral carbapenems, CS-834 and L-084, on the rat and mouse caecal microflora. AB - We determined the influence of a new quinolone, DU-6859a, and two oral carbapenems, CS-834 and L-084, on the rat and mouse caecal microflora. The caecum skin fistula-implanted rats and conventional mice were given oral antimicrobials at doses of 30 mg/kg bid for 5 days. DU-6859a generated a marked decrease in the numbers of caecal flora except for enterococci. CS-834 and L-084 had little impact on the rat caecal flora. CS-834 caused a great decrease in the numbers of mouse caecal flora but L-084 did not. In vitro studies suggest that the difference is due to the extension of inactivation of antimicrobials by caecum contents. PMID- 11062208 TI - In vivo activity of levofloxacin alone or in combination with imipenem or amikacin in a mouse model of Acinetobacter baumannii pneumonia. AB - We evaluated the in vivo activity of levofloxacin alone or in combination with imipenem or amikacin in a mouse model of Acinetobacter baumannii pneumonia using a susceptible strain and one with low-level resistance (MIC/MBC of levofloxacin: 0.06/0.06 and 4/4 mg/L, respectively). As demonstrated previously with other pathogens, the AUC/MIC ratio predicted the efficacy of fluoroquinolones against A. baumannii. This parameter correlated with bactericidal effect and survival. Combination therapy did not enhance the efficacy of levofloxacin. PMID- 11062209 TI - Effect of a novel mucoadhesive polysaccharide obtained from tamarind seeds on the intraocular penetration of gentamicin and ofloxacin in rabbits. AB - This report describes the efficacy of a novel mucoadhesive polymer, the tamarind seed polysaccharide, as a delivery system for the ocular administration of hydrophilic and hydrophobic antibiotics. Healthy rabbits were subjected to repeated ocular instillations with either conventional gentamicin or ofloxacin or these agents viscosified with the tamarind seed polysaccharide. Administration of viscosified preparations produced antibiotic concentrations both in the aqueous humour and cornea that were significantly higher than those achieved with the drugs alone. The increased drug absorption and the prolonged drug elimination phase obtained with the viscosified formulations indicate the usefulness of the tamarind seed polysaccharide as an ophthalmic delivery system for topical administration of antibiotics. PMID- 11062210 TI - A novel method for collecting and detecting amoxycillin in urine: a tool for testing antibiotic compliance in the community. AB - Assessing compliance to prescribed antibiotics in community studies of respiratory tract infections is difficult. We describe a simple method for collecting and detecting amoxycillin in urine using urine dip-sticks in conjunction with a bioassay. Urine was collected at timed intervals from eight healthy volunteers following oral amoxycillin administration. Dip-sticks inoculated with urine collected 1 and 8 h after antibiotic resulted in mean zones of inhibition of 1.75 and 1.37 cm, respectively. Amoxycillin activity remained demonstrable 14 days after inoculation of dip-sticks with urine. Dip-sticks inoculated with urine from control subjects who had not taken amoxycillin did not cause inhibition. PMID- 11062211 TI - Pharmacokinetics of a parenteral carbapenem, biapenem, in patients with end-stage renal disease and influence of haemodialysis. AB - The effects of haemodialysis on the pharmacokinetics of a carbapenem, biapenem, were evaluated in five patients with end-stage renal disease, who received 1 h iv infusions of 300 mg biapenem on both the days on and off 4 h haemodialysis. With haemodialysis, plasma biapenem exhibited two elimination phases, one during and the other after haemodialysis with half-lives of 1.16 +/- 0.12 and 3.33 +/- 0. 91 h, respectively. Ninety percent of biapenem was removed from blood to dialysate. Without haemodialysis, plasma biapenem was mono-exponentially eliminated with a half-life of 4.35 +/- 1.30 h. PMID- 11062212 TI - Safety and efficacy of recombinant granulocyte colony-stimulating factor as an adjunctive therapy for Streptococcus pneumoniae meningitis in non-neutropenic adult patients: a pilot study. AB - Twenty-two non-neutropenic adult patients with Streptococcus pneumoniae meningitis received granulocyte-colony stimulating factor (G-CSF) (300-450 Ig/day subcutaneously for 6 days) in addition to cefotaxime plus dexamethasone (9-12 g/day for 10 days and 16 mg/day for 3 days iv, respectively). Patients recovered without evident sequelae in all cases but one (with bilateral hearing deficit). No adverse event was recorded. Improvement of inflammation indices in the cerebrospinal fluid was rapid. The most rapid improvement was seen in glucose concentration, which returned to normal ranges within 24-48 h of treatment. In this study G-CSF administration appeared to be safe and effective; further controlled clinical trials are justified. PMID- 11062213 TI - Quinolone-induced QT interval prolongation: a not-so-unexpected class effect. PMID- 11062214 TI - Examination of base pair variants of the strA-strB streptomycin resistance genes from bacterial pathogens of humans, animals and plants. PMID- 11062215 TI - Use of mastalex to detect methicillin resistance in coagulase-negative staphylococci. PMID- 11062216 TI - Effect of medium composition on the MIC breakpoint for gentamicin. PMID- 11062217 TI - Activity of the tea component epicatechin gallate and analogues against methicillin-resistant Staphylococcus aureus. PMID- 11062218 TI - An in vitro investigation of the antimicrobial activity of oxifulvic acid. PMID- 11062219 TI - In vitro activity of gatifloxacin against Streptococcus pneumoniae isolates in Germany. PMID- 11062220 TI - In vitro activity of beta-lactams against gentamicin-susceptible and gentamicin resistant Staphylococcus aureus. PMID- 11062221 TI - Activity of nine antimicrobial agents against clinical isolates of Klebsiella pneumoniae producing extended-spectrum beta-lactamases and deficient or not in porins. PMID- 11062222 TI - Resistance to ceftriaxone and cefotaxime in non-typhoidal Salmonella enterica in England and Wales, 1998-99. PMID- 11062223 TI - The emergence of resistance to levofloxacin before clinical use in a university hospital in Singapore. PMID- 11062224 TI - Terminology standards for nursing: collaboration at the summit. AB - OBJECTIVE: The objective of the 1999 Nursing Vocabulary Summit Conference was to seek consensus on and a common approach to the development of nursing terminology standards for use in information systems. METHODS: A four-day invitational conference brought together authors and representatives of responsible organizations concerned with the nursing terminologies recognized or under consideration by the American Nurses Association, along with experts on language and standards and representatives of professional organizations, federal agencies, and the health informatics industry. RESULTS: Participants distinguished between colloquial terminologies and reference terminologies, and between information models and terminology models. They agreed that most recognized nursing terminologies were colloquial terminologies and that a reference terminology was needed. They formed task forces to develop and test aspects of a reference terminology model prior to a second meeting in June 2000, at which they would determine readiness to collaborate on a single international standard. DISCUSSION: The 1999 Nursing Vocabulary Summit Conference changed the level of discussion about nursing vocabulary standards from a debate about the relative merits of the various terminologies recognized in the United States to an examination of methods for developing and testing a reference terminology model and, eventually, a reference terminology that could serve as an international standard. PMID- 11062225 TI - Standards for nursing terminology. AB - Terminology work in nursing has given rise to an increasing number of nursing terminologies. These generally take the form of controlled vocabularies. Because of the limitations of the controlled vocabulary approach, individual terminologies tend to be tuned to meet the specific needs of their intended users. Differences between terminologies are now a significant barrier to the comparison and interchange of health information. To agree on a single, multipurpose terminology would be problematic. However, several options for resolving unnecessary differences between nursing terminologies are currently being explored by international standards bodies and other groups, such as the U.S. Nursing Vocabulary Summit. One such option is the use of a terminology model to facilitate evolution toward a more coherent range of terminologies. The authors describe the motivation behind the development of a standard for nursing terminologies. They explain how a terminology model might form the basis for such a standard through a description of the approach taken by CEN TC251 (the Health Informatics Technical Committee of the European Committee for Standardization). They also discuss possible limitations of standardization. PMID- 11062226 TI - Evaluation of the clinical LOINC (Logical Observation Identifiers, Names, and Codes) semantic structure as a terminology model for standardized assessment measures. AB - OBJECTIVE: The purpose of this study was to test the adequacy of the Clinical LOINC (Logical Observation Identifiers, Names, and Codes) semantic structure as a terminology model for standardized assessment measures. METHODS: After extension of the definitions, 1, 096 items from 35 standardized assessment instruments were dissected into the elements of the Clinical LOINC semantic structure. An additional coder dissected at least one randomly selected item from each instrument. When multiple scale types occurred in a single instrument, a second coder dissected one randomly selected item representative of each scale type. RESULTS: The results support the adequacy of the Clinical LOINC semantic structure as a terminology model for standardized assessments. Using the revised definitions, the coders were able to dissect into the elements of Clinical LOINC all the standardized assessment items in the sample instruments. Percentage agreement for each element was as follows: component, 100 percent; property, 87.8 percent; timing, 82.9 percent; system/sample, 100 percent; scale, 92.6 percent; and method, 97.6 percent. DISCUSSION: This evaluation was an initial step toward the representation of standardized assessment items in a manner that facilitates data sharing and re-use. Further clarification of the definitions, especially those related to time and property, is required to improve inter-rater reliability and to harmonize the representations with similar items already in LOINC. PMID- 11062227 TI - Embedded structures and representation of nursing knowledge. AB - Nursing Vocabulary Summit participants were challenged to consider whether reference terminology and information models might be a way to move toward better capture of data in electronic medical records. A requirement of such reference models is fidelity to representations of domain knowledge. This article discusses embedded structures in three different approaches to organizing domain knowledge: scientific reasoning, expertise, and standardized nursing languages. The concept of pressure ulcer is presented as an example of the various ways lexical elements used in relation to a specific concept are organized across systems. Different approaches to structuring information-the clinical information system, minimum data sets, and standardized messaging formats-are similarly discussed. Recommendations include identification of the polyhierarchies and categorical structures required within a reference terminology, systematic evaluations of the extent to which structured information accurately and completely represents domain knowledge, and modifications or extensions to existing multidisciplinary efforts. PMID- 11062228 TI - Health professionals' views of informatics education: findings from the AMIA 1999 spring conference. AB - Health care leaders emphasize the need to include information technology and informatics concepts in formal education programs, yet integration of informatics into health educational programs has progressed slowly. The AMIA 1999 Spring Congress was held to address informatics educational issues across health professions, including the educational needs in the various health professions, goals for health informatics education, and implementation strategies to achieve these goals. This paper presents the results from AMIA work groups focused on informatics education for non-informatics health professionals. In the categories of informatics needs, goals, and strategies, conference attendees suggested elements in these areas: educational responsibilities for faculty and students, organizational responsibilities, core computer skills and informatics knowledge, how to learn informatics skills, and resources required to implement educational strategies. PMID- 11062229 TI - Advanced Technology Program's information infrastructure for healthcare focused program: a brief history. AB - This review examines how a "bottom-up" model of a civilian technology program works by recounting the story of the "genesis" of the Information Infrastructure for Healthcare (IIH) focused program of the Advanced Technology Program. The IIH program began with an exchange of ideas among members of the private and public sectors (through the submission of "white papers" by members of industry, workshops conducted by the ATP, and meetings among persons from both groups) to identify the technologies that are necessary for the development of a national information infrastructure in health care. What the ATP hoped to gain through this process and how the private sector responded are noted. A statistical description of the participants and a discussion of changes in the ATP review and selection process is included. PMID- 11062230 TI - Termination of a contract to implement an enterprise electronic medical record system. AB - The development of integrated health care systems, the building of distributed computer networks throughout them, and the advent of easy-to-use electronic medical records for ambulatory practices combine to create a powerful argument for an enterprise electronic medical record. Potential customers need to learn from both successes and failures. Although the author could find in the literature only two reports of failures, a survey of family practice residencies revealed ten programs in which use of an electronic medical record had been discontinued. The author reports on a project that was terminated even though the technology was adequate to achieve the original project goals. PMID- 11062231 TI - Impact of a computer-based patient record system on data collection, knowledge organization, and reasoning. AB - OBJECTIVE: To assess the effects of a computer-based patient record system on human cognition. Computer-based patient record systems can be considered "cognitive artifacts," which shape the way in which health care workers obtain, organize, and reason with knowledge. DESIGN: Study 1 compared physicians' organization of clinical information in paper-based and computer-based patient records in a diabetes clinic. Study 2 extended the first study to include analysis of doctor-patient-computer interactions, which were recorded on video in their entirety. In Study 3, physicians' interactions with computer-based records were followed through interviews and automatic logging of cases entered in the computer-based patient record. RESULTS: Results indicate that exposure to the computer-based patient record was associated with changes in physicians' information gathering and reasoning strategies. Differences were found in the content and organization of information, with paper records having a narrative structure, while the computer-based records were organized into discrete items of information. The differences in knowledge organization had an effect on data gathering strategies, where the nature of doctor-patient dialogue was influenced by the structure of the computer-based patient record system. CONCLUSION: Technology has a profound influence in shaping cognitive behavior, and the potential effects of cognition on technology design needs to be explored. PMID- 11062232 TI - Automated mapping of observation codes using extensional definitions. AB - OBJECTIVE: To create "extensional definitions" of laboratory codes from derived characteristics of coded values in a clinical database and then use these definitions in the automated mapping of codes between disparate facilities. DESIGN: Repository data for two laboratory facilities in the Intermountain Health Care system were analyzed to create extensional definitions for the local codes of each facility. These definitions were then matched using automated matching software to create mappings between the shared local codes. The results were compared with the mappings of the vocabulary developers. MEASUREMENTS: The number of correct matches and the size of the match group were recorded. A match was considered correct if the corresponding codes from each facility were included in the group. The group size was defined as the total number of codes in the match group (e.g., a one-to-one mapping is a group size of two). RESULTS: Of the matches generated by the automated matching software, 81 percent were correct. The average group size was 2.4. There were a total of 328 possible matches in the data set, and 75 percent of these were correctly identified. CONCLUSIONS: Extensional definitions for local codes created from repository data can be utilized to automatically map codes from disparate systems. This approach, if generalized to other systems, can reduce the effort required to map one system to another while increasing mapping consistency. PMID- 11062233 TI - Automatic detection of acute bacterial pneumonia from chest X-ray reports. AB - OBJECTIVE: To evaluate the performance of a natural language processing system in extracting pneumonia-related concepts from chest x-ray reports. METHODS: DESIGN: Four physicians, three lay persons, a natural language processing system, and two keyword searches (designated AAKS and KS) detected the presence or absence of three pneumonia-related concepts and inferred the presence or absence of acute bacterial pneumonia from 292 chest x-ray reports. Gold standard: Majority vote of three independent physicians. Reliability of the gold standard was measured. OUTCOME MEASURES: Recall, precision, specificity, and agreement (using Finn's R: statistic) with respect to the gold standard. Differences between the physicians and the other subjects were tested using the McNemar test for each pneumonia concept and for the disease inference of acute bacterial pneumonia. RESULTS: Reliability of the reference standard ranged from 0.86 to 0.96. Recall, precision, specificity, and agreement (Finn R:) for the inference on acute bacterial pneumonia were, respectively, 0.94, 0.87, 0.91, and 0.84 for physicians; 0.95, 0.78, 0.85, and 0.75 for natural language processing system; 0.46, 0.89, 0.95, and 0.54 for lay persons; 0.79, 0.63, 0.71, and 0.49 for AAKS; and 0.87, 0.70, 0.77, and 0.62 for KS. The McNemar pairwise comparisons showed differences between one physician and the natural language processing system for the infiltrate concept and between another physician and the natural language processing system for the inference on acute bacterial pneumonia. The comparisons also showed that most physicians were significantly different from the other subjects in all pneumonia concepts and the disease inference. CONCLUSION: In extracting pneumonia related concepts from chest x-ray reports, the performance of the natural language processing system was similar to that of physicians and better than that of lay persons and keyword searches. The encoded pneumonia information has the potential to support several pneumonia-related applications used in our institution. The applications include a decision support system called the antibiotic assistant, a computerized clinical protocol for pneumonia, and a quality assurance application in the radiology department. PMID- 11062234 TI - The usefulness of dynamically categorizing search results. AB - OBJECTIVE: The authors' goal was to determine whether dynamic categorization, a new technique for organizing search results, is more useful than the two existing organizational techniques: relevance ranking and clustering. They define a useful tool as one that helps users learn about the kinds of information that pertain to their query, find answers to their questions efficiently and easily, and feel satisfied with their search experience. DESIGN: Fifteen patients with breast cancer and their family members completed query-related tasks using all three tools. The authors measured the time it took the subjects to accomplish their tasks, the number of answers to the query that the subjects found in four minutes, and the number of new answers that they could recall at the end of the study. Subjects also completed a user-satisfaction questionnaire. RESULTS: The results showed that patients with breast cancer and their family members could find significantly (P: < 0.05) more answers in a fixed amount of time and were significantly (P: < 0.05) more satisfied with their search experience when they used the dynamic categorization tool than when they used either the cluster tool or the ranking tool. Subjects indicated that the dynamic categorization tool provided an organization of search results that was more clear, easy to use, accurate, precise, and helpful than those of the other tools. CONCLUSION: The experiments indicate that dynamic categorization is an effective and useful approach for organizing search results. Tools that use this technique will help patients and their families gain quick and easy access to important medical information. PMID- 11062235 TI - The forkhead-associated domain of NBS1 is essential for nuclear foci formation after irradiation but not essential for hRAD50[middle dot]hMRE11[middle dot]NBS1 complex DNA repair activity. AB - NBS1 (p95), the protein responsible for Nijmegen breakage syndrome, shows a weak homology to the yeast Xrs2 protein at the N terminus region, known as the forkhead-associated (FHA) domain and the BRCA1 C terminus domain. The protein interacts with hMRE11 to form a complex with a nuclease activity for initiation of both nonhomologous end joining and homologous recombination. Here, we show in vivo direct evidence that NBS1 recruits the hMRE11 nuclease complex into the cell nucleus and leads to the formation of foci by utilizing different functions from several domains. The amino acid sequence at 665-693 on the C terminus of NBS1, where a novel identical sequence with yeast Xrs2 protein was found, is essential for hMRE11 binding. The hMRE11-binding region is necessary for both nuclear localization of the complex and for cellular radiation resistance. On the other hand, the FHA domain regulates nuclear foci formation of the multiprotein complex in response to DNA damage but is not essential for nuclear transportation of the complex and radiation resistance. Because the FHA/BRCA1 C terminus domain is widely conserved in eukaryotic nuclear proteins related to the cell cycle, gene regulation, and DNA repair, the foci formation could be associated with many phenotypes of Nijmegen breakage syndrome other than radiation sensitivity. PMID- 11062236 TI - The molecular chaperone DnaJ is required for the degradation of a soluble abnormal protein in Escherichia coli. AB - In addition to promoting protein folding and translocation, molecular chaperones of Hsp70/DnaJ families are essential for the selective breakdown of many unfolded proteins. It has been proposed that chaperones function in degradation to maintain the substrates in a soluble form. In Escherichia coli, a nonsecreted alkaline phosphatase mutant that lacks its signal sequence (PhoADelta2-22) fails to fold in the cytosol and is rapidly degraded at 37 degrees C. We show that PhoADelta2-22 is degraded by two ATP-dependent proteases, La (Lon) and ClpAP, and breakdown by both is blocked in a dnaJ259-ts mutant at 37 degrees C. Both proteases could be immunoprecipitated with PhoA, but to a much lesser extent in the dnaJ mutant. Therefore, DnaJ appears to promote formation of protease substrate complexes. DnaJ could be coimmunoprecipitated with PhoA, and the extent of this association directly correlated with its rate of degradation. Although PhoA was not degraded when DnaJ was inactivated, 50% or more of the PhoA remained soluble. PhoA breakdown and solubility did not require ClpB. PhoA degradation was reduced in a thioredoxin-reductase mutant (trxB), which allowed PhoADelta2-22 to fold into an active form in the cytosol. Introduction of the dnaJ mutation into trxB cells further stabilized PhoA, increased enzyme activity, and left PhoA completely soluble. Thus, DnaJ, although not necessary for folding (or preventing PhoA aggregation), is required for PhoA degradation and must play an active role in this process beyond maintaining the substrate in a soluble form. PMID- 11062237 TI - N-methyl D-aspartate receptor-mediated bidirectional control of extracellular signal-regulated kinase activity in cortical neuronal cultures. AB - N-Methyl D-aspartate (NMDA) receptor activation of extracellular-signal regulated kinase (ERK) was examined in primary cortical cultures. Tetrodotoxin, NMDA receptor antagonists, or reduced extracellular calcium (0.1 mm) greatly decreased basal levels of phospho-ERK2, indicating that activity-dependent activation of NMDA receptors maintained a high level of basal ERK2 activation. This activity dependent activation of phospho-ERK2 was blocked by pertussis toxin and inhibition of calcium/calmodulin-dependent kinase II and phosphatidylinositol 3 kinase but not by inhibition of protein kinase C or cAMP-dependent protein kinase. Addition of a calcium ionophore or 100 microm NMDA decreased phospho-ERK2 in the presence of 1 mm extracellular calcium but enhanced phospho-ERK2 in 0.1 mm extracellular calcium. The reduction in basal phospho-ERK2 by 100 microm NMDA was also reflected as a decrease in phospho-cAMP response element-binding protein. Inhibition of tyrosine phosphatases and serine/threonine phosphatases protein phosphatase 1 (PP1), PP2A, and PP2B did not prevent the inhibitory effect of NMDA. In the presence of tetrodotoxin, NMDA produced a bell-shaped dose-response curve with stimulation of phospho-ERK2 at 10, 25, and 50 microm NMDA and reduced stimulation at 100 microm NMDA. NMDA (50 microm) stimulation of phospho-ERK2 was completely blocked by pertussis toxin and inhibitors of phosphatidylinositol 3 kinase and was partially blocked by a calcium/calmodulin-dependent kinase II inhibitor. These results suggests that NMDA receptors can bidirectionally control ERK signaling. PMID- 11062238 TI - Ras-related GTPase Rhob represses NF-kappaB signaling. AB - rhoB encoding a Ras-related GTPase is immediate-early inducible by genotoxic treatments, indicating that it is part of the cellular stress response. Here, we investigated the influence of RhoB on signal pathways that are rapidly evoked by genotoxic compounds. The data obtained show that wild-type RhoB neither affects activation of mitogen-activated protein kinases nor AP-1-dependent gene expression. However, RhoB inhibited both basal and genotoxic agent-stimulated activity of the transcription factor nuclear factor kappaB (NF-kappaB). Thus, RhoB attenuated alkylation-induced increase in the DNA binding activity of NF kappaB and abrogated NF-kappaB-driven gene expression. Furthermore, RhoB inhibited decrease in the cellular amount of IkappaBalpha after genotoxic stress as well as after tumor necrosis factor alpha and 12-O-tetradecanoylphorbol acetate treatment. This indicates that RhoB represses NF-kappaB activation by inhibiting dissociation and subsequent degradation of IkappaBalpha. On the basis of the data, we suggest that RhoB is a novel negative regulator of NF-kappaB signaling. PMID- 11062239 TI - Nuclear factor-kappa B activation by the CXC chemokine melanoma growth stimulatory activity/growth-regulated protein involves the MEKK1/p38 mitogen activated protein kinase pathway. AB - Melanoma growth stimulatory activity/growth-regulated protein (MGSA/GRO), a CXC chemokine, plays an important role in inflammation, wound healing, growth regulation, angiogenesis, and tumorigenesis. Constitutive expression of MGSA/GROalpha in melanoma tumors is associated with constitutive nuclear factor (NF)-kappaB activity. We show here that either exogenous addition or continuous expression of MGSA/GROalpha in immortalized melanocytes enhances NF-kappaB activation, as well as mitogen-activated protein (MAP) kinase kinase kinase (MEKK) 1, MAP kinase kinase (MEK) 3/6, and p38 MAP kinase activation. Expression of dominant negative M-Ras (S27N), dominant negative MEKK1 (K432M), or specific chemical inhibitors for p38 MAP kinase (SB202190 and SB203580) block MGSA/GROalpha-induced NF-kappaB transactivation, demonstrating that Ras, MEKK1, and p38 are involved in the signal pathways of MGSA/GROalpha activation of NF kappaB. Expression of dominant active Ras or dominant active MEKK1 alone can also stimulate NF-kappaB activation. The expression of dominant negative MEKK1 inhibits the Ras-induced NF-kappaB activation, suggesting that MEKK1 is a downstream target of Ras. Moreover, MGSA/GROalpha induction of NF-kappaB is independent of the MEK1/ERK cascade, because MGSA/GROalpha failed to increase ERK and ELK activation, and specific chemical inhibitors for MEK1 (PD98059) had no effect on MGSA/GROalpha-enhanced NF-kappaB activation. These data demonstrate that NF-kappaB activation is required for MGSA/GROalpha-induced melanocyte transformation through a Ras/MEKK1/p38 cascade in melanocytes. PMID- 11062240 TI - The effect of transcription and translation initiation frequencies on the stochastic fluctuations in prokaryotic gene expression. AB - The kinetics of prokaryotic gene expression has been modelled by the Monte Carlo computer simulation algorithm of Gillespie, which allowed the study of random fluctuations in the number of protein molecules during gene expression. The model, when applied to the simulation of LacZ gene expression, is in good agreement with experimental data. The influence of the frequencies of transcription and translation initiation on random fluctuations in gene expression has been studied in a number of simulations in which promoter and ribosome binding site effectiveness has been changed in the range of values reported for various prokaryotic genes. We show that the genes expressed from strong promoters produce the protein evenly, with a rate that does not vary significantly among cells. The genes with very weak promoters express the protein in "bursts" occurring at random time intervals. Therefore, if the low level of gene expression results from the low frequency of transcription initiation, huge fluctuations arise. In contrast, the protein can be produced with a low and uniform rate if the gene has a strong promoter and a slow rate of ribosome binding (a weak ribosome binding site). The implications of these findings for the expression of regulatory proteins are discussed. PMID- 11062241 TI - Inhibition of the calcium-dependent tyrosine kinase (CADTK) blocks monocyte spreading and motility. AB - Freshly isolated peripheral blood monocytes lack focal adhesion kinase (p125(FAK)) but activate a second member of this kinase family, calcium-dependent tyrosine kinase (CADTK; also known as Pyk2/CAKbeta/RAFTK/FAK2), upon adhesion or stimulation with chemokines. To study the role of CADTK in monocyte adherence and motility, we performed immunocytochemical localization that showed CADTK at the leading edge and ruffling lamellipodial structures in freshly isolated, adhered human monocytes. We next introduced CADTK/CAKbeta-related non-kinase (CRNK), the C-terminal noncatalytic domain of CADTK, into monocytes by electroporation and showed that it inhibited CADTK autophosphorylation. Introduction of the fusion protein glutathione S-transferase (GST)-CRNK also reduced (i) cell spreading, as reflected in a reduced cell area 30 min after adhesion, (ii) adhesion-induced phosphotyrosine increases and redistribution into lamellipodia, and (iii) adhesion-induced extracellular signal-regulated protein kinase (ERK) activation. In control experiments, introduction of GST or GST-C3 transferase (an inhibitor of RhoA GTPase activity) by electroporation did not affect these parameters. Monocytes adhered in the presence of autologous serum were highly motile even after introduction of GST (83% motile cells). However, only 26% of monocytes with introduced GST-CRNK were motile. In contrast, GST-CRNK-treated monocytes were fully capable of phagocytosis and adhesion-induced cytokine gene induction, suggesting that CADTK is not involved in these cellular activities and that GST CRNK introduction does not inhibit global monocyte functions. These results suggest that CADTK is crucial for the in vitro monocyte cytoskeletal reorganization necessary for cell motility and is likely to be required in vivo for recruitment to sites of inflammation. PMID- 11062242 TI - Core histone acetylation is regulated by linker histone stoichiometry in vivo. AB - We investigated the relationship between linker histone stoichiometry and the acetylation of core histones in vivo. Exponentially growing cell lines induced to overproduce either of two H1 variants, H1(0) or H1c, displayed significantly reduced rates of incorporation of [(3)H]acetate into all four core histones. Pulse-chase experiments indicated that the rates of histone deacetylation were similar in all cell lines. These effects were also observed in nuclei isolated from these cells upon labeling with [(3)H]acetyl-CoA. Nuclear extracts prepared from control and H1-overexpressing cell lines displayed similar levels of histone acetylation activity on chromatin templates prepared from control cells. In contrast, extracts prepared from control cells were significantly less active on chromatin templates prepared from H1-overexpressing cells than on templates prepared from control cells. Reduced levels of acetylation in H1-overproducing cell lines do not appear to depend on higher order chromatin structure, because it persists even after digestion of the chromatin with micrococcal nuclease. The results suggest that alterations in chromatin structure, resulting from changes in linker histone stoichiometry may modulate the levels or rates of core histone acetylation in vivo. PMID- 11062243 TI - Residues within the conserved helicase motifs of UL9, the origin-binding protein of herpes simplex virus-1, are essential for helicase activity but not for dimerization or origin binding activity. AB - UL9, an essential gene for herpes simplex virus type 1 (HSV-1) DNA replication, exhibits helicase and origin DNA binding activities. It has been hypothesized that UL9 binds and unwinds the HSV-1 origin of replication, creating a replication bubble and promoting the assembly of the viral replication machinery; however, direct confirmation of this hypothesis has not been possible. Based on the presence of conserved helicase motifs, UL9 has been classified as a superfamily II helicase. Mutations in conserved residues of the helicase motifs I VI of UL9 have been isolated, and most of them fail to complement a UL9 null virus in vivo (Martinez R., Shao L., and Weller S. (1992) J. Virol. 66, 6735 6746). In addition, mutants in motifs I, II, and VI were found to be transdominant (Malik, A. K., and Weller, S. K. (1996) J. Virol. 70, 7859-7866). Here we present the characterization of the biochemical properties of the UL9 helicase motif mutants. We report that mutations in motifs I-IV and VI affect the ATPase activity, and all but the motif III mutation completely abolish the helicase activity. In addition, mutations in these motifs do not interfere with UL9 dimerization or the ability of UL9 to bind the HSV-1 origin of replication. Based on the similarity of the helicase motif sequences between UL9 and UvrB, another superfamily II member with helicase-like activity, we were able to map the UL9 mutations on the structure of the UvrB protein and provide an explanation for the observed phenotypes. Our results indicate that the helicase function of UL9 is indispensable for viral replication, supporting the hypothesis that UL9 is essential for unwinding the HSV-1 origin of replication in vivo. Furthermore, the data presented provide insights into the mechanism of transdominance of the UL9 helicase motif mutants. PMID- 11062244 TI - An activation switch in the ligand binding pocket of the C5a receptor. AB - Although agonists are thought to occupy binding pockets within the seven-helix core of serpentine receptors, the topography of these binding pockets and the conformational changes responsible for receptor activation are poorly understood. To identify the ligand binding pocket in the receptor for complement factor 5a (C5aR), we assessed binding affinities of hexapeptide ligands, each mutated at a single position, for seven mutant C5aRs, each mutated at a single position in the putative ligand binding site. In ChaW (an antagonist) and W5Cha (an agonist), the side chains at position 5 are tryptophan and cyclohexylalanine, respectively. Comparisons of binding affinities indicated that the hexapeptide residue at this position interacts with two C5aR residues, Ile-116 (helix III) and Val-286 (helix VII); in a C5aR model these two side chains point toward one another. Both the I116A and the V286A mutations markedly increased binding affinity of W5Cha but not that of ChaW. Moreover, ChaW, the antagonist hexapeptide, acted as a full agonist on the I116A mutant. These results argue that C5aR residues Ile-116 and Val-286 interact with the side chain at position 5 of the hexapeptide ligand to form an activation switch. Based on this and previous work, we present a docking model for the hexapeptide within the C5aR binding pocket. We propose that agonists induce a small change in the relative orientations of helices III and VII and that these helices work together to allow movement of helix VI away from the receptor core, thereby triggering G protein activation. PMID- 11062245 TI - Pleiotropic effects of Pasteurella multocida toxin are mediated by Gq-dependent and -independent mechanisms. involvement of Gq but not G11. AB - Pasteurella multocida toxin (PMT) is a highly potent mitogen for a variety of cell types. PMT has been shown to induce various cellular signaling processes, and it has been suggested to function through the heterotrimeric G-proteins G(q)/G(11). To analyze the role of G(q)/G(11) in the action of PMT, we have studied the effect of the toxin in Galpha(q)/Galpha(11) double-deficient fibroblasts as well as in fibroblasts lacking only Galpha(q) or Galpha(11). Interestingly, formation of inositol phosphates in response to PMT was exclusively dependent on Galpha(q) but not on the closely related Galpha(11). Although Galpha(q)/Galpha(11) double-deficient and Galpha(q)-deficient cells did not respond with any production of inositol phosphates to PMT, PMT was still able to induce various other cellular effects in these cells, including the activation of Rho, the Rho-dependent formation of actin stress fibers and focal adhesions, as well as the stimulation of c-Jun N-terminal kinase and extracellular signal regulated kinase. These data show that PMT leads to a variety of cellular effects that are mediated only in part by the heterotrimeric G-protein G(q). PMID- 11062246 TI - Translesion DNA synthesis by yeast DNA polymerase eta on templates containing N2 guanine adducts of 1,3-butadiene metabolites. AB - Yeast DNA polymerase eta can replicate through cis-syn cyclobutane pyrimidine dimers and 8-oxoguanine lesions with the same efficiency and accuracy as replication of an undamaged template. Previously, it has been shown that Escherichia coli DNA polymerases I, II, and III are incapable of bypassing DNA substrates containing N(2)-guanine adducts of stereoisomeric 1,3-butadiene metabolites. Here we showed that yeast polymerase eta replicates DNA containing the monoadducts (S)-butadiene monoepoxide and (S,S)-butadiene diolepoxide N(2) guanines albeit at an approximately 200-300-fold lower efficiency relative to the control guanine. Interestingly, nucleotide incorporation opposite the (R) butadiene monoepoxide and the (R,R)-butadiene diolepoxide N(2)-guanines was approximately 10-fold less efficient than incorporation opposite their S stereoisomers. Polymerase eta preferentially incorporates the correct nucleotide opposite and downstream of all four adducts, except that it shows high misincorporation frequencies for elongation of C paired with (R)-butadiene monoepoxide N(2)-guanine. Additionally, polymerase eta does not bypass the (R,R)- and (S,S)-butadiene diolepoxide N(2)-guanine-N(2)-guanine intra- strand cross links, and replication is completely blocked just prior to the lesion. Collectively, these data suggest that polymerase eta can tolerate the geometric distortions in DNA conferred by the N(2)-guanine butadiene monoadducts but not the intrastrand cross-links. PMID- 11062247 TI - Biophysical characterization of the cocaine binding pocket in the serotonin transporter using a fluorescent cocaine analogue as a molecular reporter. AB - To explore the biophysical properties of the binding site for cocaine and related compounds in the serotonin transporter SERT, a high affinity cocaine analogue (3beta-(4-methylphenyl)tropane-2beta-carboxylic acid N-(N-methyl-N-(4-nitrobenzo 2-oxa-1,3-diazol-7-yl)ethanolamine ester hydrochloride (RTI-233); K(I) = 14 nm) that contained the environmentally sensitive fluorescent moiety 7-nitrobenzo-2 oxa-1,3-diazole (NBD) was synthesized. Specific binding of RTI-233 to the rat serotonin transporter, purified from Sf-9 insect cells, was demonstrated by the competitive inhibition of fluorescence using excess serotonin, citalopram, or RTI 55 (2beta-carbomethoxy-3beta-(4-iodophenyl)tropane). Moreover, specific binding was evidenced by measurement of steady-state fluorescence anisotropy, showing constrained mobility of bound RTI-233 relative to RTI-233 free in solution. The fluorescence of bound RTI-233 displayed an emission maximum (lambda(max)) of 532 nm, corresponding to a 4-nm blue shift as compared with the lambda(max) of RTI 233 in aqueous solution and corresponding to the lambda(max) of RTI-233 in 80% dioxane. Collisional quenching experiments revealed that the aqueous quencher potassium iodide was able to quench the fluorescence of RTI-233 in the binding pocket (K(SV =) 1.7 m(-)(1)), although not to the same extent as free RTI-233 (K(SV =) 7.2 m(-)(1)). Conversely, the hydrophobic quencher 2,2,6,6 tetramethylpiperidine-N-oxyl (TEMPO) quenched the fluorescence of bound RTI-233 more efficiently than free RTI-233. These data are consistent with a highly hydrophobic microenvironment in the binding pocket for cocaine-like uptake inhibitors. However, in contrast to what has been observed for small-molecule binding sites in, for example, G protein-coupled receptors, the bound cocaine analogue was still accessible for aqueous quenching and, thus, partially exposed to solvent. PMID- 11062248 TI - Molecular cloning and characterization of the human protein kinase D2. A novel member of the protein kinase D family of serine threonine kinases. AB - We have isolated the full-length cDNA of a novel human serine threonine protein kinase gene. The deduced protein sequence contains two cysteine-rich motifs at the N terminus, a pleckstrin homology domain, and a catalytic domain containing all the characteristic sequence motifs of serine protein kinases. It exhibits the strongest homology to the serine threonine protein kinases PKD/PKCmicro and PKCnu, particularly in the duplex zinc finger-like cysteine-rich motif, in the pleckstrin homology domain and in the protein kinase domain. In contrast, it shows only a low degree of sequence similarity to other members of the PKC family. Therefore, the new protein has been termed protein kinase D2 (PKD2). The mRNA of PKD2 is widely expressed in human and murine tissues. It encodes a protein with a molecular mass of 105 kDa in SDS-polyacrylamide gel electrophoresis, which is expressed in various human cell lines, including HL60 cells, which do not express PKCmicro. In vivo phorbol ester binding studies demonstrated a concentration-dependent binding of [(3)H]phorbol 12,13-dibutyrate to PKD2. The addition of phorbol 12,13-dibutyrate in the presence of dioleoylphosphatidylserine stimulated the autophosphorylation of PKD2 in a synergistic fashion. Phorbol esters also stimulated autophosphorylation of PKD2 in intact cells. PKD2 activated by phorbol esters efficiently phosphorylated the exogenous substrate histone H1. In addition, we could identify the C-terminal Ser(876) residue as an in vivo phosphorylation site within PKD2. Phosphorylation of Ser(876) of PKD2 correlated with the activation status of the kinase. Finally, gastrin was found to be a physiological activator of PKD2 in human AGS-B cells stably transfected with the CCK(B)/gastrin receptor. Thus, PKD2 is a novel phorbol ester- and growth factor-stimulated protein kinase. PMID- 11062249 TI - Light-induced reorganization of phospholipids in rod disc membranes. AB - The transbilayer redistribution of spin-labeled phospholipid analogues (SL-PL) with choline, serine, and ethanolamine head groups (PC, PS, and PE, respectively) was studied on intact disc vesicles of bovine rod outer segment membranes in the dark and after illumination. Redistribution was measured by the extraction of spin-labeled lipid analogues from the outer leaflet of membrane using the bovine serum albumin back-exchange assay. In the dark, PS was distributed asymmetrically, favoring the outer leaflet, whereas PC and PE showed small if any asymmetry. Green illumination for 1 min caused lipid head group-specific reorganization of SL-PL. Extraction of SL-PS by bovine serum albumin showed a fast transient (<10 min) enhancement, which was further augmented by a peptide stabilizing the active metarhodopsin II conformation. The data suggest a direct release of 1 molecule of bound PS per rhodopsin into the outer leaflet and subsequent redistribution between the two leaflets. SL-PE and SL-PC showed more complex kinetics, in both cases consistent with a prolonged period of reduced extraction (2 phospholipids per rhodopsin in each case). The different phases of SL-PL reorganization after illumination may be related to the formation and decay of the active rhodopsin species and to their subsequent regeneration process. PMID- 11062250 TI - Combinatorial mRNA regulation: iron regulatory proteins and iso-iron-responsive elements (Iso-IREs). PMID- 11062251 TI - Akt regulates cell survival and apoptosis at a postmitochondrial level. AB - Phosphoinositide 3 kinase/Akt pathway plays an essential role in neuronal survival. However, the cellular mechanisms by which Akt suppresses cell death and protects neurons from apoptosis remain unclear. We previously showed that transient expression of constitutively active Akt inhibits ceramide-induced death of hybrid motor neuron 1 cells. Here we show that stable expression of either constitutively active Akt or Bcl-2 inhibits apoptosis, but only Bcl-2 prevents the release of cytochrome c from mitochondria, suggesting that Akt regulates apoptosis at a postmitochondrial level. Consistent with this, overexpressing active Akt rescues cells from apoptosis without altering expression levels of endogenous Bcl-2, Bcl-x, or Bax. Akt inhibits apoptosis induced by microinjection of cytochrome c and lysates from cells expressing active Akt inhibit cytochrome c induced caspase activation in a cell-free assay while lysates from Bcl-2 expressing cells have no effect. Addition of cytochrome c and dATP to lysates from cells expressing active Akt do not activate caspase-9 or -3 and immunoprecipitated Akt added to control lysates blocks cytochrome c-induced activation of the caspase cascade. Taken together, these data suggest that Akt inhibits activation of caspase-9 and -3 by posttranslational modification of a cytosolic factor downstream of cytochrome c and before activation of caspase-9. PMID- 11062252 TI - DRAL is a p53-responsive gene whose four and a half LIM domain protein product induces apoptosis. AB - DRAL is a four and a half LIM domain protein identified because of its differential expression between normal human myoblasts and the malignant counterparts, rhabdomyosarcoma cells. In the current study, we demonstrate that transcription of the DRAL gene can be stimulated by p53, since transient expression of functional p53 in rhabdomyosarcoma cells as well as stimulation of endogenous p53 by ionizing radiation in wild-type cells enhances DRAL mRNA levels. In support of these observations, five potential p53 target sites could be identified in the promoter region of the human DRAL gene. To obtain insight into the possible functions of DRAL, ectopic expression experiments were performed. Interestingly, DRAL expression efficiently triggered apoptosis in three cell lines of different origin to the extent that no cells could be generated that stably overexpressed this protein. However, transient transfection experiments as well as immunofluorescence staining of the endogenous protein allowed for the localization of DRAL in different cellular compartments, namely cytoplasm, nucleus, focal contacts, as well as Z-discs and to a lesser extent the M-bands in cardiac myofibrils. These data suggest that downregulation of DRAL might be involved in tumor development. Furthermore, DRAL expression might be important for heart function. PMID- 11062253 TI - Evidence for segregation of sphingomyelin and cholesterol during formation of COPI-coated vesicles. AB - In higher eukaryotes, phospholipid and cholesterol synthesis occurs mainly in the endoplasmic reticulum, whereas sphingomyelin and higher glycosphingolipids are synthesized in the Golgi apparatus. Lipids like cholesterol and sphingomyelin are gradually enriched along the secretory pathway, with their highest concentration at the plasma membrane. How a cell succeeds in maintaining organelle-specific lipid compositions, despite a steady flow of incoming and outgoing transport carriers along the secretory pathway, is not yet clear. Transport and sorting along the secretory pathway of both proteins and most lipids are thought to be mediated by vesicular transport, with coat protein I (COPI) vesicles operating in the early secretory pathway. Although the protein constituents of these transport intermediates are characterized in great detail, much less is known about their lipid content. Using nano-electrospray ionization tandem mass spectrometry for quantitative lipid analysis of COPI-coated vesicles and their parental Golgi membranes, we find only low amounts of sphingomyelin and cholesterol in COPI coated vesicles compared with their donor Golgi membranes, providing evidence for a significant segregation from COPI vesicles of these lipids. In addition, our data indicate a sorting of individual sphingomyelin molecular species. The possible molecular mechanisms underlying this segregation, as well as implications on COPI function, are discussed. PMID- 11062254 TI - Autophagic tubes: vacuolar invaginations involved in lateral membrane sorting and inverse vesicle budding. AB - Many intracellular compartments of eukaryotic cells do not adopt a spherical shape, which would be expected in the absence of mechanisms organizing their structure. However, little is known about the principles determining the shape of organelles. We have observed very defined structural changes of vacuoles, the lysosome equivalents of yeast. The vacuolar membrane can form a large tubular invagination from which vesicles bud off into the lumen of the organelle. Formation of the tube is regulated via the Apg/Aut pathway. Its lumen is continuous with the cytosol, making this inverse budding reaction equivalent to microautophagocytosis. The tube is highly dynamic, often branched, and defined by a sharp kink of the vacuolar membrane at the site of invagination. The tube is formed by vacuoles in an autonomous fashion. It persists after vacuole isolation and, therefore, is independent of surrounding cytoskeleton. There is a striking lateral heterogeneity along the tube, with a high density of transmembrane particles at the base and a smooth zone devoid of transmembrane particles at the tip where budding occurs. We postulate a lateral sorting mechanism along the tube that mediates a depletion of large transmembrane proteins at the tip and results in the inverse budding of lipid-rich vesicles into the lumen of the organelle. PMID- 11062255 TI - Cell-free reconstitution of microautophagic vacuole invagination and vesicle formation. AB - Many organelles change their shape in the course of the cell cycle or in response to environmental conditions. Lysosomes undergo drastic changes of shape during microautophagocytosis, which include the invagination of their boundary membrane and the subsequent scission of vesicles into the lumen of the organelle. The mechanism driving these structural changes is enigmatic. We have begun to analyze this process by reconstituting microautophagocytosis in a cell-free system. Isolated yeast vacuoles took up fluorescent dyes or reporter enzymes in a cytosol , ATP-, and temperature-dependent fashion. During the uptake reaction, vacuolar membrane invaginations, called autophagic tubes, were observed. The reaction resulted in the transient formation of autophagic bodies in the vacuolar lumen, which were degraded upon prolonged incubation. Under starvation conditions, the system reproduced the induction of autophagocytosis and depended on specific gene products, which were identified in screens for mutants deficient in autophagocytosis. Microautophagic uptake depended on the activity of the vacuolar ATPase and was sensitive to GTPgammaS, indicating a requirement for GTPases and for the vacuolar membrane potential. However, microautophagocytosis was independent of known factors for vacuolar fusion and vesicular trafficking. Therefore, scission of the invaginated membrane must occur via a novel mechanism distinct from the homotypic fusion of vacuolar membranes. PMID- 11062256 TI - Epidermal growth factor and membrane trafficking. EGF receptor activation of endocytosis requires Rab5a. AB - Activated epidermal growth factor receptors recruit various intracellular proteins leading to signal generation and endocytic trafficking. Although activated receptors are rapidly internalized into the endocytic compartment and subsequently degraded in lysosomes, the linkage between signaling and endocytosis is not well understood. Here we show that EGF stimulation of NR6 cells induces a specific, rapid and transient activation of Rab5a. EGF also enhanced translocation of the Rab5 effector, early endosomal autoantigen 1 (EEA1), from cytosol to membrane. The activation of endocytosis, fluid phase and receptor mediated, by EGF was enhanced by Rab5a expression, but not by Rab5b, Rab5c, or Rab5a truncated at the NH(2) and/or COOH terminus. Dominant negative Rab5a (Rab5:N34) blocked EGF-stimulated receptor-mediated and fluid-phase endocytosis. EGF activation of Rab5a function was dependent on tyrosine residues in the COOH terminal domain of the EGF receptor (EGFR). Removal of the entire COOH terminus by truncation (c'973 and c'991) abrogated ligand-induced Rab5a activation of endocytosis. A "kinase-dead" EGFR failed to stimulate Rab5a function. However, another EGF receptor mutant (c'1000), with the kinase domain intact and a single autophosphorylation site effectively signaled Rab5 activation. These results indicate that EGFR and Rab5a are linked via a cascade that results in the activation of Rab5a and that appears essential for internalization. The results point to an interdependent relationship between receptor activation, signal generation and endocytosis. PMID- 11062257 TI - New component of the vacuolar class C-Vps complex couples nucleotide exchange on the Ypt7 GTPase to SNARE-dependent docking and fusion. AB - The class C subset of vacuolar protein sorting (Vps) proteins (Vps11, Vps18, Vps16 and Vps33) assembles into a vacuole/prevacuole-associated complex. Here we demonstrate that the class C-Vps complex contains two additional proteins, Vps39 and Vps41. The COOH-terminal 148 amino acids of Vps39 direct its association with the class C-Vps complex by binding to Vps11. A previous study has shown that a large protein complex containing Vps39 and Vps41 functions as a downstream effector of the active, GTP-bound form of Ypt7, a rab GTPase required for the fusion of vesicular intermediates with the vacuole (Price, A., D. Seals, W. Wickner, and C. Ungermann. 2000. J. Cell Biol. 148:1231-1238). Here we present data that indicate that this complex also functions to stimulate nucleotide exchange on Ypt7. We show that Vps39 directly binds the GDP-bound and nucleotide free forms of Ypt7 and that purified Vps39 stimulates nucleotide exchange on Ypt7. We propose that the class C-Vps complex both promotes Vps39-dependent nucleotide exchange on Ypt7 and, based on the work of Price et al., acts as a Ypt7 effector that tethers transport vesicles to the vacuole. Thus, the class C Vps complex directs multiple reactions during the docking and fusion of vesicles with the vacuole, each of which contributes to the overall specificity and efficiency of this transport process. PMID- 11062258 TI - Cytokeratins 8 and 19 in the mouse placental development. AB - To investigate the expression and biological roles of cytokeratin 19 (K19) in development and in adult tissues, we inactivated the mouse K19 gene (Krt1-19) by inserting a bacterial beta-galactosidase gene (lacZ) by homologous recombination in embryonic stem cells, and established germ line mutant mice. Both heterozygous and homozygous mutant mice were viable, fertile, and appeared normal. By 7.5-8.0 days post coitum (dpc), heterozygous mutant embryos expressed lacZ in the notochordal plate and hindgut diverticulum, reflecting the fact that the notochord and the gut endoderm are derived from the axial mesoderm-originated cells. In the adult mutant, lacZ was expressed mainly in epithelial tissues. To investigate the possible functional cooperation and synergy between K19 and K8, we then constructed compound homozygous mutants, whose embryos died approximately 10 dpc. The lethality resulted from defects in the placenta where both K19 and K8 are normally expressed. As early as 9. 5 dpc, the compound mutant placenta had an excessive number of giant trophoblasts, but lacked proper labyrinthine trophoblast or spongiotrophoblast development, which apparently caused flooding of the maternal blood into the embryonic placenta. These results indicate that K19 and K8 cooperate in ensuring the normal development of placental tissues. PMID- 11062259 TI - Subcellular distribution of envoplakin and periplakin: insights into their role as precursors of the epidermal cornified envelope. AB - Envoplakin and periplakin are two plakins that are precursors of the epidermal cornified envelope. We studied their distribution and interactions by transfection of primary human keratinocytes and other cells. Full-length periplakin localized to desmosomes, the interdesmosomal plasma membrane and intermediate filaments. Full length envoplakin also localized to desmosomes, but mainly accumulated in nuclear and cytoplasmic aggregates with associated intermediate filaments. The envoplakin rod domain was required for aggregation and the periplakin rod domain was necessary and sufficient to redistribute envoplakin to desmosomes and the cytoskeleton, confirming earlier predictions that the proteins can heterodimerize. The linker domain of each protein was required for intermediate filament association. Like the NH(2) terminus of desmoplakin, that of periplakin localized to desmosomes; however, in addition, the periplakin NH(2) terminus accumulated at cell surface microvilli in association with cortical actin. Endogenous periplakin was redistributed from microvilli when keratinocytes were treated with the actin disrupting drug Latrunculin B. We propose that whereas envoplakin and periplakin can localize independently to desmosomes, the distribution of envoplakin at the interdesmosomal plasma membrane depends on heterodimerization with periplakin and that the NH(2) terminus of periplakin therefore plays a key role in forming the scaffold on which the cornified envelope is assembled. PMID- 11062260 TI - A selective transport route from Golgi to late endosomes that requires the yeast GGA proteins. AB - Pep12p is a yeast syntaxin located primarily in late endosomes. Using mutagenesis of a green fluorescent protein chimera we have identified a sorting signal FSDSPEF, which is required for transport of Pep12p from the exocytic pathway to late endosomes, from which it can, when overexpressed, reach the vacuole. When this signal is mutated, Pep12p instead passes to early endosomes, a step that is determined by its transmembrane domain. Surprisingly, Pep12p is then specifically retained in early endosomes and does not go on to late endosomes. By testing appropriate chimeras in mutant strains, we found that FSDSPEF-dependent sorting was abolished in strains lacking Gga1p and Gga2p, Golgi-associated coat proteins with homology to gamma adaptin. In the gga1 gga2 double mutant endogenous Pep12p cofractionated with the early endosome marker Tlg1p, and recycling of Snc1p through early endosomes was defective. Pep12p sorting was also defective in cells lacking the clathrin heavy or light chain. We suggest that specific and direct delivery of proteins to early and late endosomes is required to maintain the functional heterogeneity of the endocytic pathway and that the GGA proteins, probably in association with clathrin, help create vesicles destined for late endosomes. PMID- 11062261 TI - Rabenosyn-5, a novel Rab5 effector, is complexed with hVPS45 and recruited to endosomes through a FYVE finger domain. AB - Rab5 regulates endocytic membrane traffic by specifically recruiting cytosolic effector proteins to their site of action on early endosomal membranes. We have characterized a new Rab5 effector complex involved in endosomal fusion events. This complex includes a novel protein, Rabenosyn-5, which, like the previously characterized Rab5 effector early endosome antigen 1 (EEA1), contains an FYVE finger domain and is recruited in a phosphatidylinositol-3-kinase-dependent fashion to early endosomes. Rabenosyn-5 is complexed to the Sec1-like protein hVPS45. hVPS45 does not interact directly with Rab5, therefore Rabenosyn-5 serves as a molecular link between hVPS45 and the Rab5 GTPase. This property suggests that Rabenosyn-5 is a closer mammalian functional homologue of yeast Vac1p than EEA1. Furthermore, although both EEA1 and Rabenosyn-5 are required for early endosomal fusion, only overexpression of Rabenosyn-5 inhibits cathepsin D processing, suggesting that the two proteins play distinct roles in endosomal trafficking. We propose that Rab5-dependent formation of membrane domains enriched in phosphatidylinositol-3-phosphate has evolved as a mechanism for the recruitment of multiple effector proteins to mammalian early endosomes, and that these domains are multifunctional, depending on the differing activities of the effector proteins recruited. PMID- 11062262 TI - Pds5p is an essential chromosomal protein required for both sister chromatid cohesion and condensation in Saccharomyces cerevisiae. AB - The PDS5 gene (precocious dissociation of sisters) was identified in a genetic screen designed to identify genes important for chromosome structure. PDS5 is an essential gene and homologues are found from yeast to humans. Pds5p function is important for viability from S phase through mitosis and localizes to chromosomes during this cell cycle window, which encompasses the times when sister chromatid cohesion exists. Pds5p is required to maintain cohesion at centromere proximal and distal sequences. These properties are identical to those of the four cohesion complex members Mcd1p/Scc1p, Smc1p, Smc3p, and Scc3p/Irr1p (Guacci, V., D. Koshland, and A. Strunnikov. 1997. Cell. 91:47-57; Michaelis, C., R. Ciosk, and K. Nasmyth. 1997. Cell. 91:35-45; Toth, A., R. Ciosk, F. Uhlmann, M. Galova, A. Schleiffer, and K. Nasmyth. 1999. Genes Dev. 13:307-319). Pds5p binds to centromeric and arm sequences bound by Mcd1p. Furthermore, Pds5p localization to chromosomes is dependent on Mcd1p. Thus, Pds5p, like the cohesin complex members, is a component of the molecular glue that mediates sister chromatid cohesion. However, Mcd1p localization to chromosomes is independent of Pds5p, which may reflect differences in their roles in cohesion. Finally, Pds5p is required for condensation as well as cohesion, which confirms the link between these processes revealed through analysis of Mcd1p (Guacci, V., D. Koshland, and A. Strunnikov. 1997. Cell. 91:47-57). Therefore, the link between cohesion and condensation is a general property of yeast chromosomes. PMID- 11062263 TI - ACAPs are arf6 GTPase-activating proteins that function in the cell periphery. AB - The GTP-binding protein ADP-ribosylation factor 6 (Arf6) regulates endosomal membrane trafficking and the actin cytoskeleton in the cell periphery. GTPase activating proteins (GAPs) are critical regulators of Arf function, controlling the return of Arf to the inactive GDP-bound state. Here, we report the identification and characterization of two Arf6 GAPs, ACAP1 and ACAP2. Together with two previously described Arf GAPs, ASAP1 and PAP, they can be grouped into a protein family defined by several common structural motifs including coiled coil, pleckstrin homology, Arf GAP, and three complete ankyrin-repeat domains. All contain phosphoinositide-dependent GAP activity. ACAP1 and ACAP2 are widely expressed and occur together in the various cultured cell lines we examined. Similar to ASAP1, ACAP1 and ACAP2 were recruited to and, when overexpressed, inhibited the formation of platelet-derived growth factor (PDGF)-induced dorsal membrane ruffles in NIH 3T3 fibroblasts. However, in contrast with ASAP1, ACAP1 and ACAP2 functioned as Arf6 GAPs. In vitro, ACAP1 and ACAP2 preferred Arf6 as a substrate, rather than Arf1 and Arf5, more so than did ASAP1. In HeLa cells, overexpression of either ACAP blocked the formation of Arf6-dependent protrusions. In addition, ACAP1 and ACAP2 were recruited to peripheral, tubular membranes, where activation of Arf6 occurs to allow membrane recycling back to the plasma membrane. ASAP1 did not inhibit Arf6-dependent protrusions and was not recruited by Arf6 to tubular membranes. The additional effects of ASAP1 on PDGF induced ruffling in fibroblasts suggest that multiple Arf GAPs function coordinately in the cell periphery. PMID- 11062264 TI - D-Titin: a giant protein with dual roles in chromosomes and muscles. AB - Previously, we reported that chromosomes contain a giant filamentous protein, which we identified as titin, a component of muscle sarcomeres. Here, we report the sequence of the entire titin gene in Drosophila melanogaster, D-Titin, and show that it encodes a two-megadalton protein with significant colinear homology to the NH(2)-terminal half of vertebrate titin. Mutations in D-Titin cause chromosome undercondensation, chromosome breakage, loss of diploidy, and premature sister chromatid separation. Additionally, D-Titin mutants have defects in myoblast fusion and muscle organization. The phenotypes of the D-Titin mutants suggest parallel roles for titin in both muscle and chromosome structure and elasticity, and provide new insight into chromosome structure. PMID- 11062265 TI - Mutational analysis of fibrillarin and its mobility in living human cells. AB - Cajal bodies (CBs) are subnuclear organelles that contain components of a number of distinct pathways in RNA transcription and RNA processing. CBs have been linked to other subnuclear organelles such as nucleoli, but the reason for the presence of nucleolar proteins such as fibrillarin in CBs remains uncertain. Here, we use full-length fibrillarin and truncated fibrillarin mutants fused to green fluorescent protein (GFP) to demonstrate that specific structural domains of fibrillarin are required for correct intranuclear localization of fibrillarin to nucleoli and CBs. The second spacer domain and carboxy terminal alpha-helix domain in particular appear to target fibrillarin, respectively, to the nucleolar transcription centers and CBs. The presence of the RNP domain seems to be a prerequisite for correct targeting of fibrillarin. Time-lapse confocal microscopy of human cells that stably express fibrillarin-GFP shows that CBs fuse and split, albeit at low frequencies. Recovered fluorescence of fibrillarin-GFP in nucleoli and CBs after photobleaching indicates that it is highly mobile in both organelles (estimated diffusion constant approximately 0.02 microm(2) s(-1)), and has a significantly larger mobile fraction in CBs than in nucleoli. PMID- 11062266 TI - Matching of calcineurin activity to upstream effectors is critical for skeletal muscle fiber growth. AB - Calcineurin-dependent pathways have been implicated in the hypertrophic response of skeletal muscle to functional overload (OV) (Dunn, S.E., J.L. Burns, and R.N. Michel. 1999. J. Biol. Chem. 274:21908-21912). Here we show that skeletal muscles overexpressing an activated form of calcineurin (CnA*) exhibit a phenotype indistinguishable from wild-type counterparts under normal weightbearing conditions and respond to OV with a similar doubling in cell size and slow fiber number. These adaptations occurred despite the fact that CnA* muscles displayed threefold higher calcineurin activity and enhanced dephosphorylation of the calcineurin targets NFATc1, MEF2A, and MEF2D. Moreover, when calcineurin signaling is compromised with cyclosporin A, muscles from OV wild-type mice display a lower molecular weight form of CnA, originally detected in failing hearts, whereas CnA* muscles are spared this manifestation. We also show that OV induced growth and type transformations are prevented in muscle fibers of transgenic mice overexpressing a peptide that inhibits calmodulin from signaling to target enzymes. Taken together, these findings provide evidence that both calcineurin and its activity-linked upstream signaling elements are crucial for muscle adaptations to OV and that, unless significantly compromised, endogenous levels of this enzyme can accommodate large fluctuations in upstream calcium dependent signaling events. PMID- 11062267 TI - The dendritic cell receptor for endocytosis, DEC-205, can recycle and enhance antigen presentation via major histocompatibility complex class II-positive lysosomal compartments. AB - Many receptors for endocytosis recycle into and out of cells through early endosomes. We now find in dendritic cells that the DEC-205 multilectin receptor targets late endosomes or lysosomes rich in major histocompatibility complex class II (MHC II) products, whereas the homologous macrophage mannose receptor (MMR), as expected, is found in more peripheral endosomes. To analyze this finding, the cytosolic tails of DEC-205 and MMR were fused to the external domain of the CD16 Fcgamma receptor and studied in stable L cell transfectants. The two cytosolic domains each mediated rapid uptake of human immunoglobulin (Ig)G followed by recycling of intact CD16 to the cell surface. However, the DEC-205 tail recycled the CD16 through MHC II-positive late endosomal/lysosomal vacuoles and also mediated a 100-fold increase in antigen presentation. The mechanism of late endosomal targeting, which occurred in the absence of human IgG, involved two functional regions: a membrane-proximal region with a coated pit sequence for uptake, and a distal region with an EDE triad for the unusual deeper targeting. Therefore, the DEC-205 cytosolic domain mediates a new pathway of receptor mediated endocytosis that entails efficient recycling through late endosomes and a greatly enhanced efficiency of antigen presentation to CD4(+) T cells. PMID- 11062268 TI - Evidence that beta3 integrin-induced Rac activation involves the calpain dependent formation of integrin clusters that are distinct from the focal complexes and focal adhesions that form as Rac and RhoA become active. AB - Interaction of integrins with the extracellular matrix leads to transmission of signals, cytoskeletal reorganizations, and changes in cell behavior. While many signaling molecules are known to be activated within Rac-induced focal complexes or Rho-induced focal adhesions, the way in which integrin-mediated adhesion leads to activation of Rac and Rho is not known. In the present study, we identified clusters of integrin that formed upstream of Rac activation. These clusters contained a Rac-binding protein(s) and appeared to be involved in Rac activation. The integrin clusters contained calpain and calpain-cleaved beta3 integrin, while the focal complexes and focal adhesions that formed once Rac and Rho were activated did not. Moreover, the integrin clusters were dependent on calpain for their formation. In contrast, while Rac- and Rho-GTPases were dependent on calpain for their activation, formation of focal complexes and focal adhesions by constitutively active Rac or Rho, respectively, occurred even when calpain inhibitors were present. Taken together, these data are consistent with a model in which integrin-induced Rac activation requires the formation of integrin clusters. The clusters form in a calpain-dependent manner, contain calpain, calpain-cleaved integrin, and a Rac binding protein(s). Once Rac is activated, other integrin signaling complexes are formed by a calpain-independent mechanism(s). PMID- 11062269 TI - Localization and activity of myosin light chain kinase isoforms during the cell cycle. AB - Phosphorylation on Ser 19 of the myosin II regulatory light chain by myosin light chain kinase (MLCK) regulates actomyosin contractility in smooth muscle and vertebrate nonmuscle cells. The smooth/nonmuscle MLCK gene locus produces two kinases, a high molecular weight isoform (long MLCK) and a low molecular weight isoform (short MLCK), that are differentially expressed in smooth and nonmuscle tissues. To study the relative localization of the MLCK isoforms in cultured nonmuscle cells and to determine the spatial and temporal dynamics of MLCK localization during mitosis, we constructed green fluorescent protein fusions of the long and short MLCKs. In interphase cells, localization of the long MLCK to stress fibers is mediated by five DXRXXL motifs, which span the junction of the NH(2)-terminal extension and the short MLCK. In contrast, localization of the long MLCK to the cleavage furrow in dividing cells requires the five DXRXXL motifs as well as additional amino acid sequences present in the NH(2)-terminal extension. Thus, it appears that nonmuscle cells utilize different mechanisms for targeting the long MLCK to actomyosin structures during interphase and mitosis. Further studies have shown that the long MLCK has twofold lower kinase activity in early mitosis than in interphase or in the early stages of postmitotic spreading. These findings suggest a model in which MLCK and the myosin II phosphatase (Totsukawa, G., Y. Yamakita, S. Yamashiro, H. Hosoya, D.J. Hartshorne, and F. Matsumura. 1999. J. Cell Biol. 144:735-744) act cooperatively to regulate the level of Ser 19-phosphorylated myosin II during mitosis and initiate cytokinesis through the activation of myosin II motor activity. PMID- 11062270 TI - Chlamydomonas IFT88 and its mouse homologue, polycystic kidney disease gene tg737, are required for assembly of cilia and flagella. AB - Intraflagellar transport (IFT) is a rapid movement of multi-subunit protein particles along flagellar microtubules and is required for assembly and maintenance of eukaryotic flagella. We cloned and sequenced a Chlamydomonas cDNA encoding the IFT88 subunit of the IFT particle and identified a Chlamydomonas insertional mutant that is missing this gene. The phenotype of this mutant is normal except for the complete absence of flagella. IFT88 is homologous to mouse and human genes called Tg737. Mice with defects in Tg737 die shortly after birth from polycystic kidney disease. We show that the primary cilia in the kidney of Tg737 mutant mice are shorter than normal. This indicates that IFT is important for primary cilia assembly in mammals. It is likely that primary cilia have an important function in the kidney and that defects in their assembly can lead to polycystic kidney disease. PMID- 11062271 TI - Prm1p, a pheromone-regulated multispanning membrane protein, facilitates plasma membrane fusion during yeast mating. AB - Cell fusion occurs throughout development, from fertilization to organogenesis. The molecular mechanisms driving plasma membrane fusion in these processes remain unknown. While yeast mating offers an excellent model system in which to study cell fusion, all genes previously shown to regulate the process act at or before cell wall breakdown; i.e., well before the two plasma membranes have come in contact. Using a new strategy in which genomic data is used to predict which genes may possess a given function, we identified PRM1, a gene that is selectively expressed during mating and that encodes a multispanning transmembrane protein. Prm1p localizes to sites of cell-cell contact where fusion occurs. In matings between Deltaprm1 mutants, a large fraction of cells initiate zygote formation and degrade the cell wall separating mating partners but then fail to fuse. Electron microscopic analysis reveals that the two plasma membranes in these mating pairs are tightly apposed, remaining separated only by a uniform gap of approximately 8 nm. Thus, the phenotype of Deltaprm1 mutants defines a new step in the mating reaction in which membranes are juxtaposed, possibly through a defined adherence junction, yet remain unfused. This phenotype suggests a role for Prm1p in plasma membrane fusion. PMID- 11062272 TI - Recycling of the yeast a-factor receptor. AB - The yeast a-factor receptor (Ste3p) is subject to two mechanistically distinct modes of endocytosis: a constitutive, ligand-independent pathway and a ligand dependent uptake pathway. Whereas the constitutive pathway leads to degradation of the receptor in the vacuole, the present work finds that receptor internalized via the ligand-dependent pathway recycles. With the a-factor ligand continuously present in the culture medium, trafficking of the receptor achieves an equilibrium in which continuing uptake to endosomal compartments is balanced by its recycling return to the plasma membrane. Withdrawal of ligand from the medium leads to a net return of the internalized receptor back to the plasma membrane. Although recycling is demonstrated for receptors that lack the signal for constitutive endocytosis, evidence is provided indicating a participation of recycling in wild-type Ste3p trafficking as well: a-factor treatment both slows wild-type receptor turnover and results in receptor redistribution to intracellular endosomal compartments. Apparently, a-factor acts as a switch, diverting receptor from vacuole-directed endocytosis and degradation, to recycling. A model is presented for how the two Ste3p endocytic modes may collaborate to generate the polarized receptor distribution characteristic of mating cells. PMID- 11062273 TI - Lubeluzole in acute ischemic stroke treatment: A double-blind study with an 8 hour inclusion window comparing a 10-mg daily dose of lubeluzole with placebo. AB - BACKGROUND AND PURPOSE: This trial was a double-blind, placebo-controlled, phase III trial with an 8-hour inclusion window to assess the efficacy and safety of an intravenous loading dose of 7.5 mg followed by a daily intravenous dose of 10 mg lubeluzole for 5 days in acute ischemic stroke patients. METHODS: A total of 1786 patients were randomized: 901 to lubeluzole and 885 to placebo. Overall, 212 patients (23.5%) from the lubeluzole group and 213 (24.1%) from the placebo group discontinued the trial prematurely. In the lubeluzole group 201 patients (22.3%) discontinued because of adverse events compared with 193 patients (21.8%) in the placebo group. RESULTS: The primary population for the efficacy analysis comprised the core stroke patients (exclusion of older patients aged >75 years with severe stroke) in the 0- to 6-hour inclusion time window. The primary efficacy parameter was a 3-category functional status (Barthel Index 70 to 100/0 to 70/vegetative, dead) at week 12. In the lubeluzole group 207 patients (47.8%) were classified as mildly dependent/independent at week 12, 131 (30.3%) were moderately/severely dependent, and 95 (21.9%) were vegetative/dead. In the placebo group these numbers were 221 (54.4%), 112 (27.6%), and 73 (18.0%), respectively. Logistic regression analysis showed no statistically significant difference between the treatment groups (P:=0.162). Additionally, for none of the secondary efficacy parameters (mortality at week 12, modified Rankin score, total Barthel score) was a statistically significant difference between the lubeluzole and placebo groups obtained. There were no statistically significant differences between the 2 treatments for all treated patients, patients included within the 6 to 8-hour window, and patients with severe strokes aged >75 years. Overall, of all treated patients, 401 (22.5%) died: 203 (22.5%) in the lubeluzole group and 198 (22.4%) with placebo. Of all subjects treated, 853 (95%) on lubeluzole and 826 (93%) on placebo reported an adverse event during their treatment period or within the next 2 days after discontinuation of treatment. The most frequently observed adverse events were fever (25.9% lubeluzole; 23.4% placebo), constipation (20.2%; 19.7%), and headache (17.6%; 21.2%). Imbalances were found for atrial fibrillation (1.8% lubeluzole; 1.1% placebo) and QT prolongation (0.9%; 0.2%). CONCLUSIONS: This study failed to show an efficacy of lubeluzole in the treatment of acute stroke. On the other hand, lubeluzole treatment by the current dosage schedule was not associated with a significant safety problem. PMID- 11062274 TI - Combined intravenous and intra-arterial recombinant tissue plasminogen activator in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: A retrospective analysis was performed on 20 consecutive patients who presented with severe acute ischemic stroke and were evaluated for a combined intravenous (IV) and local intra-arterial (IA) recombinant tissue plasminogen activator (rtPA) thrombolytic approach within 3 hours of onset. METHODS: Twenty consecutive patients with carotid artery distribution strokes were evaluated and treated using a combined IV and IA rtPA approach over a 14 month period (September 1998 to October 1999). rtPA (0.6 mg/kg) was given intravenously (maximum dose 60 mg); 15% of the IV dose was given as bolus, followed by a continuous infusion over 30 minutes. A maximal IA dose, up to 0.3 mg/kg or 24 mg, whichever was less, was given over a maximum of 2 hours. IV treatment was initiated within 3 hours in 19 of 20 patients. All 20 patients underwent angiography, and 16 of 20 patients received local IA rtPA. RESULTS: The median baseline National Institutes of Health Stroke Scale (NIHSS) score for the 20 patients was 21 (range 11 to 31). The median time from stroke onset to IV treatment was 2 hours and 2 minutes, and median time to initiation of IA treatment was 3 hours and 30 minutes. Ten patients (50%) recovered to a modified Rankin Scale (mRS) of 0 or 1; 3 patients (15%), to an mRS of 2; and 5 patients (25%), to an mRS of 4 or 5. One patient (5%) developed a symptomatic intracerebral hemorrhage and eventually died. One other patient (5%) expired because of complications from the stroke. CONCLUSIONS: We believe that the greater-than-expected proportion of favorable outcomes in these patients with severe ischemic stroke reflects the short time to initiation of both IV and IA thrombolysis. PMID- 11062275 TI - Prediction of functional outcome and in-hospital mortality after admission with oral anticoagulant-related intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: Early survival of patients with intracerebral hemorrhage in general is known to be most strongly dependent on the Glasgow Coma Scale score on admission. The aim of this study was to examine the factors determining functional outcome and in-hospital mortality of patients admitted with an intracerebral hemorrhage related to oral anticoagulant (OAC) use. METHODS: Correlation studies and multiple logistic regression analyses were performed on data from a retrospective series of 42 patients admitted with OAC-related intracerebral hemorrhages over a 6-year period to a tertiary care center in the north of Scotland. RESULTS: The functional outcome after an OAC-related intracerebral hemorrhage was dependent on maximum diameter of hematoma on CT scan (R:=-0.72, P:<0. 001) and international normalized ratio (INR) (R:=-0.35, P:=0.024). Hematoma diameter and INR were not themselves strongly correlated (R:=0.31, P:=0.099). In-hospital mortality can be predicted by the Glasgow Coma Scale score alone (R:(2)=0.36, overall predictive accuracy 68%) but more accurately by a logistic regression model including hematoma diameter and CT signs of cerebrovascular disease (R:(2)=0.70, predictive accuracy 83%). CONCLUSIONS: Neither functional outcome nor in-hospital mortality appears to be strongly dependent on INR measured on admission. CT scan, however, provides essential information and allows accurate predictions about the short-term outcome of OAC-related intracerebral hemorrhages. PMID- 11062276 TI - Safety and cost of low-molecular-weight heparin as bridging anticoagulant therapy in subacute cerebral ischemia. AB - BACKGROUND AND PURPOSE: Anticoagulation with intravenous unfractionated heparin (IVUH) while awaiting therapeutic oral anticoagulant levels is a common practice in patients with acute and subacute cerebral ischemia. A promising alternative strategy is to use bridging subcutaneous low-molecular-weight heparin (LMWH), which may have a favorable risk-benefit profile compared with IVUH and may permit earlier discharge with completion of transition to warfarin therapy as an outpatient. METHODS: A LMWH, enoxaparin 1 mg/kg BID, was used as bridging anticoagulation therapy in 24 consecutive patients admitted to a university stroke center in whom the treatment plan included transition from acute to chronic anticoagulation. The LMWH group was contrasted with the preceding 24 patients transitioned to warfarin with IVUH at the same center. RESULTS: Fewer patients in the LMWH bridging therapy group experienced neurological worsening than in the IVUH bridging therapy group (2/24 versus 8/24; P:=0.033). Fewer total adverse events were noted in the LMWH group than in the IVUH cohort (3 versus 20; P:=0. 002). Fifteen of the 24 LMWH patients (62.5%) were discharged while still receiving LMWH and completed transition to warfarin as outpatients, receiving an average of 3.6 days of outpatient transitional therapy. In these 15 patients, use of LMWH was associated with a net savings of $2197 per patient. CONCLUSIONS: In this pilot cohort with subacute cerebral ischemia, bridging LMWH appeared to be safer than bridging IVUH and was associated with reduced hospital stay and reduced total cost of care. PMID- 11062277 TI - Resource utilization and costs of stroke unit care integrated in a care continuum: A 1-year controlled, prospective, randomized study in elderly patients: the Goteborg 70+ Stroke Study. AB - BACKGROUND AND PURPOSE: The aim of the present study was to examine resource utilization during a 12-month period after acute stroke in elderly patients randomized to care in an acute stroke unit integrated with a care continuum compared with conventional care in general medical wards. A secondary aim was to describe costs related to the severity of stroke. METHODS: Two hundred forty-nine consecutive patients aged >/=70 years with acute stroke within 7 days before admission, living in their own homes in Goteborg, Sweden, without recognized need of care were randomized to 2 groups: 166 patients were assigned to nonintensive stroke unit care with a care continuum, and 83 patients were assigned to conventional care. There was no difference in mortality or the proportion of patients living at home after 1 year. Main outcomes were costs from inpatient care, outpatient care, and informal care. RESULTS: Mean annual cost per patient was 170, 000 Swedish crowns (SEK) (equivalent to $25,373) and 191,000 SEK ($28,507) in the stroke unit and the general medical ward groups, respectively (P:=NS). Seventy percent of the total cost was for inpatient care, and 30% was for outpatient and informal care. For patients with mild, moderate, and severe stroke, the mean annual costs per patient were 107,000 SEK ($15,970), 263,000 SEK ($39, 254), and 220,000 SEK ($32,836), respectively (P:<0.001). There was no statistical difference in age or nonstroke diagnosis. CONCLUSIONS: The total costs the first year did not differ significantly between the treatment groups in this prospective study. The total annual cost per patient showed a very large variation, which was related to stroke severity at onset and not to age or nonstroke diagnoses. Costs other than those for hospital care constituted a substantial fraction of total costs and must be taken into account when organizing the management of stroke patients. The high variability in costs necessitates a larger study to assess long-term cost effectiveness. PMID- 11062278 TI - Effect of acute stroke unit care integrated with care continuum versus conventional treatment: A randomized 1-year study of elderly patients: the Goteborg 70+ Stroke Study. AB - BACKGROUND AND PURPOSE: The aim of the study was to compare the effect of conventional treatment with the effect of acute stroke unit care integrated with geriatric stroke unit care continuum. METHODS: A 1-year study was undertaken with 2:1 randomization to stroke unit care or conventional care, with assessment by an independent team. The study was composed of 249 elderly patients (aged >/=70 years) hospitalized for acute stroke, without previous cerebral lesion and without recognized need of care. Main outcome measures were patients at home after 1 year, ability in daily living activities, health-related quality of life score according to questionnaire, death or institutional care, and death or dependence. RESULTS: One hundred two patients (61%) in the stroke unit and 49 patients (59%) in the general ward group were alive and at home after 1 year (95% CI -10% to 16%). There were no significant differences in daily life activities or quality of life. In patients with concomitant cardiac disease, there was a reduction in death or institutional care after 3 months in the stroke unit group compared with the group receiving conventional care (28% versus 49%, respectively; 95% CI -40% to -3%). This effect did not remain after 1 year. Patients seeking care after 24 hours often had mild stroke and lived alone. CONCLUSIONS: There was no effect on the number of patients living at home after 1 year, but after 3 months of stroke unit care, a beneficial effect was found on mortality and the need for institutional care among those with concomitant heart disease. This study involved patients who were considerably older than those investigated in previous randomized studies of acute stroke unit care; thus, these findings will contribute to the specialized register of controlled trials in stroke. PMID- 11062279 TI - Prehospital and emergency department delays after acute stroke: the Genentech Stroke Presentation Survey. AB - BACKGROUND AND PURPOSE: Patient delays in seeking treatment for stroke and delays within the Emergency Department (ED) are major factors in the lack of use of thrombolytic therapy for stroke. The Genentech Stroke Presentation Survey was a multicentered prospective registry of patients with acute stroke. The study was designed to characterize prehospital delays and delays within the ED. METHODS: Patients with stroke symptoms presenting to 48 EDs participating in a clinical trial of acute stroke therapy were enrolled prospectively. A 1-page data form was completed from patient interviews and medical records. RESULTS: A total of 1207 subjects were entered into the study. Ninety-four percent of the 721 subjects with complete data had a diagnosis of stroke or transient ischemic attack, 13% were black, 50% were female, and 67% were aged >65 years. The median time from symptom onset to ED arrival was 2.6 (interquartile range 1.2 to 6.3) hours. The median time from ED arrival until CT scan completion was 1.1 (0.7 to 1.8) hours, and the total delay time (symptom onset until CT scan completion) had a median of 4.0 (2.3 to 8.3) hours. Patients who arrived by emergency medical services had significantly shorter prehospital delay times and times to CT scan. Age, race, sex, and educational level did not appear to affect prehospital delay times. CONCLUSIONS: Despite its limitations, this large geographically diverse study strongly suggests that the use of emergency medical services is an important modifiable determinant of delay time for the treatment of acute stroke. PMID- 11062280 TI - Determinants of use of emergency medical services in a population with stroke symptoms: the Second Delay in Accessing Stroke Healthcare (DASH II) Study. AB - BACKGROUND AND PURPOSE: With the advent of time-dependent thrombolytic therapy for ischemic stroke, it has become increasingly important for stroke patients to arrive at the hospital quickly. This study investigates the association between the use of emergency medical services (EMS) and delay time among individuals with stroke symptoms and examines the predictors of EMS use. METHODS: The Second Delay in Accessing Stroke Healthcare Study (DASH II) was a prospective study of 617 individuals arriving at emergency departments in Denver, Colo, Chapel Hill, NC, and Greenville, SC, with stroke symptoms. RESULTS: EMS use was associated with decreased prehospital and in-hospital delay. Those who used EMS had a median prehospital delay time of 2.85 hours compared with 4.03 hours for those who did not use EMS (P:=0.002). Older individuals were more likely to use EMS (odds ratio [OR] 1.21 for each 5-year increase, 95% CI 1.14 to 1.29), as were individuals who expressed a high sense of urgency about their symptoms (OR 1.69, 95% CI 1.09 to 2.62). Knowledge of stroke symptoms was not associated with increased EMS use (OR 0.63, 95% CI 0.40 to 0.98). Patients were more likely to use EMS if someone other than the patient first identified that there was a problem (OR 2.35, 95% CI 1.61 to 3.44). CONCLUSIONS: Interventions aimed at increasing EMS use among stroke patients need to stress the urgency of stroke symptoms and the importance of calling 911 and need to be broad-based, encompassing not only those at high risk for stroke but also their friends and family. PMID- 11062281 TI - Is early ischemic lesion volume on diffusion-weighted imaging an independent predictor of stroke outcome? A multivariable analysis. AB - BACKGROUND AND PURPOSE: The heterogeneity of stroke makes outcome prediction difficult. Neuroimaging parameters may improve the predictive value of clinical measures such as the National Institutes of Health Stroke Scale (NIHSS). We investigated whether the volume of early ischemic brain lesions assessed with diffusion-weighted imaging (DWI) was an independent predictor of functional outcome. METHODS: We retrospectively selected patients with nonlacunar ischemic stroke in the anterior circulation from 4 prospective Stanford Stroke Center studies evaluating early MRI. The baseline NIHSS score and ischemic stroke risk factors were assessed. A DWI MRI was performed within 48 hours of symptom onset. Clinical characteristics and early lesion volume on DWI were compared between patients with an independent outcome (Barthel Index score >/=85) and a dependent outcome (Barthel Index score <85) at 1 month. A logistic regression model was performed with factors that were significantly different between the 2 groups in univariate analysis. RESULTS: Sixty-three patients fulfilled the entry criteria. One month after symptom onset, 24 patients had a Barthel Index score <85 and 39 had a Barthel Index score >/=85. In univariate analysis, patients with independent outcome were younger, had lower baseline NIHSS scores, and had smaller lesion volumes on DWI. In a logistic regression model, DWI volume was an independent predictor of outcome, together with age and NIHSS score, after correction for imbalances in the delay between symptom onset and MRI. CONCLUSIONS: DWI lesion volume measured within 48 hours of symptom onset is an independent risk factor for functional independence. This finding could have implications for the design of acute stroke trials. PMID- 11062282 TI - Depression and other determinants of values placed on current health state by stroke patients: evidence from the VA Acute Stroke (VASt) study. AB - BACKGROUND AND PURPOSE: This prospective study examined the determinants of the utility (value) placed on health status among a sample of patients with acute ischemic and intracerebral hemorrhagic stroke. METHODS: Data were from the VA Acute Stroke (VASt) study, a nationwide prospective cohort of 1073 acute stroke patients admitted at any of 9 Department of Veterans Affairs Medical Center sites between April 1, 1995, and March 31, 1997. The primary outcome was the patient's health status utility as measured by the time-tradeoff method. Data were obtained by telephone interviews at 1, 6, and 12 months and by medical record review. General linear mixed modeling was used to assess the effects of social, psychological, and physical factors on patients' valuations of their current health state. The analysis was confined to the 327 patients who were able to provide self-reports at >/=2 time points. RESULTS: Patients' valuations of their health state status over the initial 12 months after stroke were very stable over time, with only a slight improvement at 6 months, followed by a slight decline at 12 months. In adjusted analyses, living alone, being institutionalized, decreased physical function, and depression were independently associated with lower levels of patient health status utility over time. CONCLUSIONS: Stroke patient health status utilities are relatively stable during the initial year after stroke. In addition to physical function, psychological health and social environment are important determinants of patient health status utility. These factors need to be considered when conducting stroke decision analyses if more accurate conclusions are to be drawn regarding preferred patterns of care. PMID- 11062283 TI - Clinical meaning of the Stroke-Adapted Sickness Impact Profile-30 and the Sickness Impact Profile-136. AB - BACKGROUND AND PURPOSE: Handicap or health-related quality of life (HRQL) measures are seldom used in stroke trials, although the importance of these measures has been stressed frequently. We studied the clinical meaning of the Stroke-Adapted Sickness Impact Profile-30 (SA-SIP30) and the original SIP136 for use in stroke research. METHODS: We included 418 patients who had had a stroke 6 months earlier. We studied the associations between the SA-SIP30 and SIP136 scores versus other frequently used outcome measures from the International Classification of Impairments, Disabilities, and Handicaps (ICIDH) (Barthel Index, Rankin Scale) and the HRQL model (health perception items, Euroqol). To interpret the continuous SA-SIP30 and SIP136 scores, we used receiver operating characteristic curve analysis with the aforementioned measures as external criteria. RESULTS: The psychosocial dimension scores of both SIP versions remained largely unexplained. The physical dimension and total scores of both SIP versions were mainly associated with the disability measures derived from the ICIDH model, as well as with the physical HRQL domains. Most patients with an SA SIP30 total score >33 or an SIP136 total score >22 had poor health profiles. There were no major differences between the SA-SIP30 and the SIP136, although the SA-SIP30 scores were less skewed toward the healthier outcomes than the SIP136. CONCLUSIONS: Our study showed that (1) both SIP total scores primarily represent aspects of physical functioning and not HRQL; (2) both SIP versions provide more clinical information than the frequently used disability measures; and (3) the SA SIP30 should be preferred over the SIP136. PMID- 11062284 TI - Incidence and risk factors for subtypes of cerebral infarction in a general population: the Hisayama study. AB - BACKGROUND AND PURPOSE: We estimated the incidence of first-ever cerebral infarction in regard to its subtypes and analyzed their risk factors separately in a community-based prospective cohort study in Japan. METHODS: Stroke-free subjects (n=1621) aged >/=40 years were followed up for 32 years from 1961. During this period, 298 cerebral infarctions occurred and were divided into 167 lacunar, 62 atherothrombotic, 56 cardioembolic, and 13 undetermined subtypes of infarction on the basis of clinical information including brain imaging and autopsy findings. RESULTS: The age-adjusted incidence of lacunar infarction (3.8 per 1000 person-years for men and 2.0 for women) was higher than that of atherothrombotic infarction (1.2, 0. 7) and cardioembolic infarction (1.3, 0.5) in both sexes. Time-dependent Cox's proportional hazard analysis revealed systolic blood pressure as well as age to be independent risk factors for all subtypes of cerebral infarction except for cardioembolic infarction in men. Additionally, ST depression on ECG, glucose intolerance, and smoking in men and left ventricular hypertrophy on ECG and body mass index in women remained significant risk factors for lacunar infarction. ST depression was also significantly related to events of atherothrombotic infarction in women. The risk of atrial fibrillation for cardioembolic infarction was outstandingly high in both sexes, and left ventricular hypertrophy and lower total cholesterol were additional risk factors for cardioembolic infarction in women. CONCLUSIONS: In this Japanese population, lacunar infarction was the most common subtype of cerebral infarction and had a greater variety of risk factors, including not only hypertension but also ECG abnormalities, diabetes, obesity, and smoking, than did atherothrombotic infarction or cardioembolic infarction. PMID- 11062285 TI - Gender differences in the risk of ischemic stroke associated with aortic atheromas. AB - BACKGROUND AND PURPOSE: Atherosclerotic plaque of the proximal portion of the aorta is associated with an increased risk of ischemic stroke in the elderly. Different cutoffs of plaque thickness have been used in the literature for risk stratification and have been applied to both men and women. However, the assumption that the relationship between plaque thickness and stroke risk is the same in the 2 genders has not been proven. The aim of this study was to evaluate whether the prevalence of different degrees of aortic plaque thickness differed in men and women with ischemic stroke. METHODS: We performed transesophageal echocardiography in 152 patients aged >59 years with acute ischemic stroke (76 men and 76 women) and in 152 control subjects of similar age (70 men and 82 women). Odds ratios (ORs) for ischemic stroke with 95% CIs for different plaque thickness definitions were calculated for the overall group and separately for men and women by logistic regression analysis after adjusting for age, arterial hypertension, and hypercholesterolemia. RESULTS: Aortic plaques >/=4 mm were significantly more frequent in men than in women (31.5% versus 20.3%, respectively; P:=0.025) and were associated with ischemic stroke in both men (adjusted OR 6.0, CI 2.1 to 16.8) and women (adjusted OR 3. 2, CI 1.2 to 8.8). However, plaques 3 to 3.9 mm in thickness had a significant association with stroke in women (adjusted OR 4.8, CI 1. 7 to 15.0) but not in men (adjusted OR 0.8, CI 0.2 to 3.0). Plaques <3 mm were not associated with a significantly increased stroke risk for either sex. CONCLUSIONS: Smaller aortic plaques are significantly associated with ischemic stroke in women but not in men. If the increased prevalence of smaller plaques in women is confirmed to be associated with increased risk for embolic stroke, different cutoff points may have to be adopted in men and women for risk stratification and for decisions regarding medical intervention. PMID- 11062286 TI - Predictors of cerebrovascular events and death among patients with valvular heart disease: A population-based study. AB - BACKGROUND AND PURPOSE: There is little population-based information on cerebrovascular events and survival among valvular heart disease patients. We used the Kaplan-Meier product-limit method and the Cox proportional hazards model to determine rates and predictors of cerebrovascular events and death among valve disease patients. METHODS: This population-based historical cohort study in Olmsted County, Minnesota, reviewed residents with a first echocardiographic diagnosis of mitral stenosis (n=19), mitral regurgitation (n=528), aortic stenosis (n=140), and aortic regurgitation (n=106) between 1985 and 1992. RESULTS: During 2694 person-years of follow-up, 98 patients developed cerebrovascular events and 356 died. Compared with expected numbers, these observations are significantly elevated, with standardized morbidity ratio of 3.2 (95% CI, 2.6 to 3.8) and 2. 5 (95% CI, 2.2 to 2.7), respectively. Independent predictors of cerebrovascular events were age, atrial fibrillation, and severe aortic stenosis. The risk ratio of severe aortic stenosis was 3.5 (95% CI, 1.4 to 8.6), with atrial fibrillation conferring greater risk at younger age. Predictors of death were age, sex, cerebrovascular events, ischemic heart disease, and congestive heart failure, the greatest risk being among those with both congestive heart failure and cerebrovascular events (risk ratio=8.8; 95% CI, 5. 8 to 13.4). Valve disease type and severity were not independent determinants of death. CONCLUSIONS: The risk of cerebrovascular events and death among patients with valve disease remains high. Age, atrial fibrillation, and severe aortic stenosis are independent predictors of cerebrovascular events, and age, sex, cerebrovascular events, congestive heart failure, and ischemic heart disease are independent predictors of death in these patients. PMID- 11062287 TI - Effect of contralateral carotid artery stenosis on carotid ultrasound velocity measurements. AB - BACKGROUND AND PURPOSE: Carotid ultrasonography is being increasingly performed as the sole investigation to assess internal carotid artery (ICA) stenosis. A potential source of error in using ultrasound peak systolic velocity (PSV) measurements is that the redistribution of blood flow due to severe stenosis in a contralateral carotid artery may lead to artificially elevated values. METHODS: Ultrasonography was performed before and after carotid endarterectomy in symptomatic patients who participated in the North American Symptomatic Carotid Endarterectomy Trial (NASCET). The mean change in PSV in the unoperated artery was assessed across all degrees of angiographically defined stenosis. A simple theoretical resistance model of the cerebral circulation was also derived. RESULTS: Complete bilateral ultrasound examinations were performed within 90 days of the initial scan in 386 patients. In the presence of a contralateral severe (70% to 99%) ICA stenosis, the PSV in the unoperated artery was artificially elevated by a mean of 84 cm/s (P:=0.03; 95% CI, 10 to 159 cm/s). The mean elevation was less pronounced for lesser degrees of stenosis (11 to 21 cm/s). Small elevations (3 to 12 cm/s) were observed when the contralateral artery had <70% stenosis. The patterns of observed results were congruent with those from the theoretical model. CONCLUSIONS: The present study showed that a severely stenosed contralateral ICA can artificially elevate ultrasound PSV. Since the effect was greatest when bilaterally severe stenoses were present, caution must be exercised when assessing the degree of ICA stenosis on the basis of ultrasonography PSV measurements alone. PMID- 11062288 TI - Use of transcranial Doppler ultrasound to predict outcome in patients with intracranial large-artery occlusive disease. AB - BACKGROUND AND PURPOSE: Intracranial large-artery occlusive disease is the predominant vascular lesion found in stroke patients of Asian, African, and Hispanic ancestry, making it numerically perhaps the most common vascular cause of stroke in the world. Relatively little is known about the clinical significance of finding such lesions. We investigate whether the presence and the extent of these vascular lesions help predict outcome after stroke. METHODS: On the basis of transcranial Doppler of the intracranial arteries with supplementary duplex ultrasound of the carotid arteries, we determined the number of occlusive arteries in the craniocervical circulation of consecutive patients who were hospitalized for acute cerebral ischemia. Patients were followed for 6 months for further vascular events (including transient ischemic attack, stroke, and acute coronary syndrome) or death. RESULTS: Among 705 consecutive Chinese patients studied, occlusive arteries were found in 345 patients (49%): 258 patients (37%) had intracranial lesions only, 71 (10%) had both extracranial and intracranial lesions, and 16 (2.3%) had extracranial lesions only. Sixty-three (18%) of the 345 patients with occlusive arteries and 35 (9.7%) of the 360 patients without occlusive arteries had further vascular event or death within 6 months. The risk of vascular events or death increased rapidly with rising numbers of occlusive arteries, after adjustment for vascular risk factors and stroke severity (adjusted odds ratio [OR] 1.25 per occlusive artery, 95% CI 1.12 to 1.39). Other independent risk factors included age (OR 1.03 per year of age, 95% CI 1.01 to 1.05) and atrial fibrillation (OR 3.00, 95% CI 1.40 to 6.69). CONCLUSIONS: In patients with predominantly intracranial large-artery occlusive disease, the presence and the total number of occlusive arteries in the craniocervical circulation predict further vascular events or death within 6 months after stroke. Transcranial Doppler ultrasound is an important investigation for the evaluation of patients with stroke in populations at risk of intracranial atherosclerotic disease. PMID- 11062289 TI - Frequencies of certain complement protein alleles and serum levels of anti-heat shock protein antibodies in cerebrovascular diseases. AB - BACKGROUND AND PURPOSE: A strong correlation exists between the intensity of atherosclerotic alterations in different arteries. Marked differences exist, however, in the age and sex distribution and risk factors for coronary heart disease (CHD) and cerebrovascular disease (CVD). We therefore performed genetic and immunologic studies in patients with CVD. METHODS: We studied 292 patients with CVD (stroke or transient ischemic attack) and as control either 198 healthy blood donors and 485 healthy elderly (aged >60 years) people (genetic study) or 94 blood donors aged 45 to 60 years and 49 healthy elderly (aged >60 years) people (anti-heat-shock protein [hsp] measurements). Allele frequencies of 3 genes (C4A, C4B, and C3) encoding proteins of the complement system were determined by electrophoresis and immunofixation. Serum concentration of autoantibodies against 60-kDa heat-shock protein (anti-hsp60) was measured by the enzyme-linked immunosorbent assay method. RESULTS: Marked differences were observed between CVD patients and controls in the genetic studies. In the CVD patients aged >60 years, the frequency (11.3%) of the deficient allele of the C4B gene (C4B*Q0) was significantly (P:=0.0003) higher than that of the healthy controls (5.4%). By contrast, in the group aged 45 to 60 years, the frequency of the C4B*Q0 allele was lower in patients than in controls. Serum concentration of anti-hsp60 in the CVD patients did not differ from control values. CONCLUSIONS: In previous studies C4B*Q0 frequency was reported to be higher in CHD patients aged 45 to 60 years than in aged-matched controls. Moreover, high anti-hsp60 levels were found in CHD patients. These findings contrast with our present report of lower frequency of C4B*Q0 in CVD patients. Therefore, genetic and immunologic factors may at least partly explain the differences between the natural history and risk factors of CHD and CVD. PMID- 11062290 TI - Analysis of endoglin expression in normal brain tissue and in cerebral arteriovenous malformations. AB - BACKGROUND AND PURPOSE: A high incidence of arteriovenous malformations (AVMs) is associated with hereditary hemorrhagic telangiectasia type 1. Endoglin, the gene mutated in this disorder, is expressed at reduced levels on blood vessels of these patients. Since endoglin is a component of the transforming growth factor beta receptor complex critical for vascular development and homeostasis, we determined its expression in sporadic cerebral AVMs and in normal brain vessels. METHODS: Twenty cerebral AVMs and 10 normal brain samples were analyzed for endoglin, platelet endothelial cell adhesion molecule 1 (PECAM-1), alpha-smooth muscle cell actin, vimentin, and desmin by immunohistochemistry. RESULTS: In normal brain, endoglin was found not only on the endothelium of all vessels but also on the adventitial layer of arteries and arterioles. In cerebral AVMs, the numerous vessels present expressed endoglin on both endothelium and adventitia. Arterialized veins, identified by lack of elastin and uneven thickness of smooth muscle cells, revealed endoglin-positive mesenchymal cells in the adventitia and perivascular connective tissue. These cells were fibroblasts since they expressed vimentin but not actin and/or desmin. CONCLUSIONS: This is the first report of endoglin expression on adventitia of normal brain arteries and on arterialized veins in cerebral AVMs. Increasing numbers of endoglin-positive endothelial and adventitial cells were seen in sporadic cerebral AVMs, but endoglin density was normal. Thus, it is not involved in the generation of these lesions. However, the presence of endoglin on fibroblasts in the perivascular stroma suggests an active role for this protein in vascular remodeling in response to increased blood flow and shear stress. PMID- 11062291 TI - Polymorphism in the promoter of lipopolysaccharide receptor CD14 and ischemic cerebrovascular disease. AB - BACKGROUND AND PURPOSE: A growing amount of evidence suggests that infectious and inflammatory processes may be involved in the initiation of arteriosclerosis, but the mechanisms are conceivably multifactorial and complex. Two European groups have recently demonstrated that a C(-260)-->T polymorphism in the promoter of the CD14 lipopolysaccharide receptor may be a risk factor for coronary artery disease (CAD). The T allele of this polymorphism reportedly increases the expression of CD14 and may be involved in atherogenesis. In the present study we investigated a possible association between the C(-260)-->T polymorphism in the CD14 promoter and the occurrence of symptomatic ischemic cerebrovascular disease (CVD). METHODS: Genotype frequencies of the C(-260)-->T polymorphism in the CD14 promoter were determined in 235 patients with CVD, as confirmed by brain CT and/or MRI, and 309 age- and sex-matched control subjects. RESULTS: The distribution of genotypes was as follows: CVD patients, T:/T: 24.3%, C:/T: 53.2%, and C:/C: 22. 6%; controls, T:/T: 26.9%, C:/T: 50.2%, and C:/C: 23.0%. There was no significant difference between the CD14 promoter genotypes of the CVD patients and the controls (chi(2)=0.601, P:=0.741). We also measured the concentration of serum soluble CD14 and the density of membranous CD14 on monocytes in the CVD patients, but the polymorphism was not associated with either the concentration of soluble CD14 or the density of membranous CD14 (P:=0.358, P:=0.238, respectively). CONCLUSIONS: Our results indicate that the C(-260)-->T polymorphism in the CD14 promoter is not associated with an increased risk for CVD. PMID- 11062292 TI - Frequency and location of microbleeds in patients with primary intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: MRI is known to detect clinically silent microbleeds (MBs) in patients with primary intracerebral hemorrhage (pICH), but the frequency and diagnostic and clinical significance of this finding are still debated. Therefore, we investigated a consecutive series of pICH patients and analyzed the patterns of MB distribution in the context of clinical variables and location of the symptomatic hematoma. METHODS: The study population consisted of 109 patients with pICH. There were 59 women and 50 men aged 22 to 91 years (mean 64.6 years). MRI was obtained on a 1.5-T system with use of a gradient-echo T2*-weighted sequence. A cohort of 280 community-dwelling asymptomatic elderly individuals who underwent the same imaging protocol served for comparison. RESULTS: MBs were seen in 59 (54%) patients and ranged in number from 1 to 90 lesions (mean 14, median 6). In the majority of patients, MBs were located simultaneously in various parts of the brain, with a preference for cortical-subcortical regions (39%) and the basal ganglia/thalami (38%). There was some tendency toward a regional association between MB location and the site of the symptomatic hematoma, but we could not discern specific patterns of MB distribution. Logistic regression analysis identified MBs, periventricular hyperintensity grades, and lacunes but not risk factors as independent variables contributing to a correct classification of pICH and control individuals. CONCLUSIONS: MBs can be detected in more than half of the patients with pICH and appear to be quite general markers of various types of bleeding-prone microangiopathy. PMID- 11062293 TI - Release of glial tissue-specific proteins after acute stroke: A comparative analysis of serum concentrations of protein S-100B and glial fibrillary acidic protein. AB - BACKGROUND AND PURPOSE: This study was aimed at the comparative analysis of serum concentrations of glial fibrillary acidic protein (GFAP) and protein S-100B in patients with acute stroke. METHODS: We investigated 32 patients with stroke symptoms consistent with cerebral ischemia in the anterior territory of vascular supply. Serial venous blood samples were taken after admission to the hospital and during the first 4 days after onset of stroke. Evaluation of lesion topography and volume of infarcted brain area was based on cranial CT data. The patients' clinical status was consecutively evaluated by the National Institutes of Health Stroke Scale (NIHSS) and the Barthel Index score at discharge from the hospital. RESULTS: Protein S-100B and GFAP release was found to be significantly correlated (r=0.96; P:<0.001). The release of both biochemical markers was associated with the volume of brain lesions (S-100B: r=0.957, P:<0.0001; GFAP: r=0.955, P:<0.0001) and the neurological status at discharge from the hospital (S 100B: r=0.821, P:=0.0002; GFAP: r=0.717, P:=0.0003). The highest correlation between both S-100B and GFAP serum concentration and Barthel score was calculated at the last time of blood sampling, 4 days after stroke onset (S-100B: r=0.621, P:<0.001; GFAP: r=0.655, P:<0.001). The release of both astroglia derived proteins differed between different subtypes of stroke. GFAP was found to be a more sensitive marker of brain damage in patients with smaller lacunar lesions or minor strokes. CONCLUSIONS: Our results indicate that postischemic release patterns of GFAP and S-100B protein may allow insight into the underlying pathophysiology of acute cerebral infarcts and may be used as a valuable tool of clinical stroke treatment. PMID- 11062294 TI - Heritability of intracerebral hemorrhagic lesions and cerebral aneurysms in the rat. AB - BACKGROUND AND PURPOSE: Under certain conditions, the Brown Norway (BN) rat is susceptible to intracerebral hemorrhagic vascular (ICV) lesions within the cerebral cortex, whereas the Long-Evans (LE) rat is prone to develop aneurysms in the circle of Willis. The incidence of these 2 pathological phenotypes was studied in progeny of different BNXLE crosses to determine their heritability in these new rat models. In addition, a possible link between ICV lesion occurrence and either the susceptibility to spontaneous rupture of the arterial internal elastic lamina (IEL) or basal plasma angiotensin-converting enzyme (ACE) activity was also studied in back-cross (BC) F1XBN rats, the only second-generation group with a high incidence of ICV lesions. METHODS: To induce cerebrovascular lesions, rats were submitted to experimental hypertension associated with ligation of 1 carotid artery. After death, the brain was examined for cerebral lesions. Numbers of arterial IEL ruptures were determined microscopically with the use of en face preparations. Plasma ACE activity was determined before the induction of hypertension. RESULTS: In general, groups that developed ICV lesions presented a low incidence of aneurysms. ICV lesion incidence was similar in F1 hybrids and BC(F1XBN) and greatly decreased in F2 and BC(F1XLE) rats compared with BN rats. No cerebral aneurysms developed in F1 rats. Aneurysmal incidence was 24% (20% ruptured) in LE, 42% (59% ruptured) in F2, and 50% (75% ruptured) in BC(F1XLE) rats. In BC(F1XBN) rats, neither the incidence of IEL rupture nor the plasma ACE activity was higher in the rats with ICV lesions. However, the mean blood pressure level was higher in these rats, and peak blood pressure was higher in rats with the most severe grades of ICV lesions. CONCLUSIONS: These data suggest a polygenic and dominant mode of inheritance of ICV pathology. The formation of aneurysms in the circle of Willis tended to be favored, and their rupture was clearly increased by the presence of BN rat alleles within the LE rat genome. These data may provide the basis for future studies to determine, in new rat models, which genes are involved in these pathologies. PMID- 11062295 TI - Stroke outcome in double-mutant antioxidant transgenic mice. AB - BACKGROUND AND PURPOSE: Both NO and superoxide cytotoxicity are important in experimental stroke; however, it is unclear whether these molecules act within parallel pathological pathways or as coreagents in a common reaction. We examined these alternatives by comparing outcomes after middle cerebral artery occlusion in male and female neuronal NO synthase (nNOS)-deficient (nNOS-/-) or human CuZn superoxide dismutase-overexpressing (hSOD1+/-) mice and a novel strain with both mutations. METHODS: Permanent middle cerebral artery occlusion was performed by use of the intraluminal filament technique (18 hours). Neurological status was scored, and tissue infarction volume was determined by 2,3,5-triphenyltetrazolium staining and image analysis. RESULTS: Hemispheric infarction volume was reduced in each transgenic strain relative to the genetically matched, wild-type, control cohorts (WT mice): nNOS-/- (80+/-6 mm(3)) and double-mutant (49+/-6 mm(3)) mice versus WT mice (114+/-7 mm(3)) and hSOD1+/- mice (52+/-7 mm(3)) versus WT mice (95+/-5 mm(3)). Human CuZn superoxide dismutase had a larger effect on mean infarction volume (30% of contralateral hemisphere) than did nNOS deficiency (46%). Although infarction volume was less in double-mutant mice compared with nNOS-/- mice, injury was not improved relative to hSOD1+/- mice. There was no difference in histological damage by sex within each strain; however, female nNOS /- mice were not protected from ischemic injury, unlike male mutants. CONCLUSIONS: Superoxide generation contributes to severe ischemic brain injury in vivo to a greater extent than does neuronally derived NO. In vivo, significant superoxide scavenging by CuZn superoxide dismutase occurs within cellular compartments or through biochemical pathways that are not restricted to, and may be distinct from, neuronal NO/superoxide reaction and peroxynitrite synthesis. PMID- 11062296 TI - 99mTc annexin V imaging of neonatal hypoxic brain injury. AB - BACKGROUND AND PURPOSE: Delayed cell loss in neonates after cerebral hypoxic ischemic injury (HII) is believed to be a major cause of cerebral palsy. In this study, we used radiolabeled annexin V, a marker of delayed cell loss (apoptosis), to image neonatal rabbits suffering from HII. METHODS: Twenty-two neonatal New Zealand White rabbits had ligation of the right common carotid artery with reduction of inspired oxygen concentration to induce HII. Experimental animals (n=17) were exposed to hypoxia until an ipsilateral hemispheric decrease in the average diffusion coefficient occurred. After reversal of hypoxia and normalization of average diffusion coefficient values, experimental animals were injected with (99m)Tc annexin V. Radionuclide images were recorded 2 hours later. RESULTS: Experimental animals showed no MR evidence of blood-brain barrier breakdown or perfusion abnormalities after hypoxia. Annexin images demonstrated multifocal brain uptake in both hemispheres of experimental but not control animals. Histology of the brains from experimental animals demonstrated scattered pyknotic cortical and hippocampal neurons with cytoplasmic vacuolization of glial cells without evidence of apoptotic nuclei by terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining. Double staining with markers of cell type and exogenous annexin V revealed that annexin V was localized in the cytoplasm of scattered neurons and astrocytes in experimental and, less commonly, control brains in the presence of an intact blood-brain barrier. CONCLUSIONS: Apoptosis may develop after HII even in brains that appear normal on diffusion-weighted and perfusion MR. These data suggest a role of radiolabeled annexin V screening of neonates at risk for the development of cerebral palsy. PMID- 11062297 TI - Estrogen decreases infarct size after temporary focal ischemia in a genetic model of type 1 diabetes mellitus. AB - BACKGROUND AND PURPOSE: It is unclear how genetic type 1 diabetes mellitus (DM) influences infarct size when blood glucose is tightly controlled. The aim of this study was to determine the effect of genetic type 1 DM, as occurs in BB rats, on infarct size after transient unilateral middle cerebral artery occlusion (MCAO) in male and female rats. In addition, studies suggest that male type 1 DM rats have a higher incidence of end-organ complications than do females. A second aim of this study was to determine the effect of chronic 17beta-estradiol (E(2)) administration on infarct size in male BB rats. METHODS: Diabetic male (MDiab, n=14) and female (FDiab, n=8) BB rats were studied and compared with background strain Wistar rats (MWist, n=16; FWist, n=14). Two additional male cohorts (MWist+E(2), n=15; MDiab+E(2), n=14) received subcutaneous 25 microg E(2) implants 7 to 10 days before MCAO. Rats underwent 1 hour of MCAO followed by 22 hours of reperfusion. Physiological variables were controlled among groups, and the intraischemic laser Doppler flow signal was reduced similarly in all animals. Infarction volume was evaluated by 2,3,5-triphenyltetrazolium chloride staining and image analysis. RESULTS: Preischemic blood glucose was 94+/-5, 127+/-13, 90+/ 15, 63+/-18, 122+/-8, and 81+/-14 mg/dL in MWist, FWist, MDiab, FDiab, MWist+E(2), and MDiab+E(2) rats, respectively (mean+/-SE). Intraischemic laser Doppler flow was reduced to 20% to 25% of baseline in all groups. Striatal infarct size (percentage of ipsilateral caudate putamen) was increased in male diabetic rats relative to nondiabetic MWist rats (41+/-3% versus 28+/-3%). Striatal injury was not increased in FDiab rats, and infarction volume was smaller than that in FWist rats (23+/-4% in FWist versus 13+/-3% in FDiab). Chronic estrogen treatment reduced cortical and striatal infarction in MDiab+E(2) rats compared with untreated MDiab rats. CONCLUSIONS: Type 1 DM is associated with increased infarct size after temporary MCAO, despite tight control of blood glucose. The deleterious effect of DM is evident only in males rats; female diabetic BB rats sustain small infarcts. Chronic E(2) treatment reduced injury in the male BB rat, providing neuroprotection even in the presence of DM. These data suggest that genetic diabetes even with mild glucose elevation plays a role in determining neuropathology in experimental stroke. However, factors such as reproductive steroids also determine outcome in DM stroke. PMID- 11062298 TI - Differences in vulnerability to permanent focal cerebral ischemia among 3 common mouse strains. AB - BACKGROUND AND PURPOSE: Genetically engineered mice are used to study the role of single genes in cerebral ischemia, but inherent, strain-dependent differences in neuronal vulnerability may affect experimental end points. To examine this possibility, tissue injury resulting from focal ischemia and its relationship to cerebral hemodynamics were determined in 3 common mutant mouse strains. METHODS: Permanent middle cerebral artery ligation was performed in male C57BL/6J, Balb/C, and 129X1/SvJ mice. Mean arterial blood pressure, blood gases, basal and postischemic cortical blood flow ([(14)C]iodoantipyrine autoradiography and laser Doppler flowmetry), posterior communicating artery patency, and infarct size were determined. RESULTS: Basal cortical blood flow did not differ among strains. Ten minutes after middle cerebral artery ligation, relative red cell flow in the ischemic cortex was 6% to 7% of preischemic flow in every strain. Despite similar hemodynamics, cortical infarcts in Balb/C mice were 3-fold larger than those in 129X1/SvJ and C57BL/6J mice; infarct size in the latter 2 strains was not significantly different. The posterior communicating artery was either poorly developed or absent in >90% of the Balb/C and C57BL/6J but in <50% of the 129X1/SvJ mice. CONCLUSIONS: The extent of ischemic injury differed markedly between the 3 strains. The presence and patency of posterior communicating arteries, although variable among strains, did not affect preischemic or postischemic cortical blood flow or bear any relationship to ischemic injury. Therefore, intrinsic factors, other than hemodynamic variability, may contribute to the differences in ischemic vulnerability among strains. These findings underscore the importance of selecting genetically matched wild-type controls. PMID- 11062299 TI - Late resolution of diffusion-weighted MRI changes in a patient with prolonged reversible ischemic neurological deficit after thrombolytic therapy. AB - BACKGROUND: Reduced apparent diffusion coefficients (ADCs) correlate with cerebral ischemia. The combination of ADC with techniques to measure cerebral perfusion may help to assess the effect of treatment. CASE DESCRIPTION: The authors describe a patient who experienced an acute stroke with hemianopia, consequently treated with intravenous recombinant tissue plasminogen activator. Positron emission tomographic imaging and MRI, including diffusion-weighted MRI, were performed during and shortly after treatment with recombinant tissue plasminogen activator and 34 to 35 hours later. Cerebral perfusion of the left occipital region was reduced to 74%. Diffusion-weighted MRI detected a territory of restricted water movement in the corresponding area. Further magnetic resonance sequences did not show any pathologies. In follow-up, positron emission tomography demonstrated reperfusion. The volume of diffusion and ADC abnormalities detected by MRI decreased markedly. A few hours later, the patient recovered completely. A third MRI examination 10 days later showed normal findings. CONCLUSIONS: In a patient with prolonged reversible ischemic neurological deficit, resolution of early diffusion changes corresponded to cerebral reperfusion and to the recovery of clinical symptoms. PMID- 11062300 TI - Cochrane report: A systematic review of mannitol therapy for acute ischemic stroke and cerebral parenchymal hemorrhage. AB - BACKGROUND: Mannitol was reported to decrease cerebral edema associated with tissue damage and is used to treat acute stroke in many countries. SUMMARY OF REVIEW: We tested whether there is any evidence from unconfounded randomized clinical trials that treatment with mannitol reduces short- and long-term case fatality and dependency if administered after ischemic stroke or cerebral parenchymal hemorrhage. Trials were identified by the standard search strategy of the Cochrane Collaboration Stroke Review Group. A supplementary MEDLINE search was performed, and the Chinese Stroke Trials Register and the Latin-American databank LILACS were checked. A search was performed of master's and PhD degree theses in the databank of Sao Paulo University and in abstracts of medical congresses on neurology and neurosurgery during 1965-1997 in Brazil. Investigators were contacted for unpublished information. Only truly randomized unconfounded clinical trials were eligible for inclusion. Two of the reviewers independently extracted data from the trials. Data synthesis and analysis was performed with the use of the Cochrane Review Manager software (RevMan version 4.0.4). CONCLUSIONS: Only 1 trial fulfilled the inclusion criteria. The number of included patients was small, and the follow-up was short. Case fatality, the proportion of dependent patients, and side effects were not reported and were not available from the investigators. As a result of lack of appropriate randomized trials, currently no conclusion can be drawn on the effects of mannitol in acute stroke. The routine use of mannitol in all patients with acute stroke is not supported by evidence from randomized controlled clinical trials. PMID- 11062301 TI - Systematic review of diffusion and perfusion imaging in acute ischemic stroke. AB - BACKGROUND AND PURPOSE: Recent advances in neuroimaging have raised hopes of early and accurate identification of ischemic brain and the discrimination of dead from salvageable tissue. We sought to determine whether the data published so far are enough to establish the roles of these techniques in everyday clinical practice. METHODS: A systematic review of studies of MR diffusion-weighted imaging (DWI), perfusion imaging (PI), or a combination of the two, in human stroke, excluding abstracts and case reports. One reviewer extracted information on the size of each study, its main purpose, methodological details, and results. RESULTS: We identified 47 studies of DWI, 18 studies of MR PI alone or in combination with another advanced imaging modality, and 19 studies of DWI and PI together. Although high proportions of the studies were prospective and gave good details of the imaging sequences used, the majority gave very limited details on patient selection and clinical characteristics or blinded imaging assessment. Pathophysiological changes were inferred from DWI/PI patterns that were not supported by other data. CONCLUSIONS: Despite considerable enthusiasm for and promise of these techniques, there is not sufficient information available in these studies to enable us to draw firm conclusions about the sensitivity and specificity of these techniques for identification of either ischemic lesions not visible by other means or salvageable tissue. Future studies should include larger numbers of carefully described patients, assess the contribution of DWI over and above other imaging, obtain follow-up at an appropriate time interval to determine accurate clinical and neuroradiological outcomes, and assess DWI/PI abnormality with reperfusion in randomized treatment trials. Investigators should also be encouraged to combine their individual patient data in meta-analyses to obtain a more robust assessment of the value of DWI and PI from larger sample sizes. PMID- 11062303 TI - Abstracts of literature PMID- 11062302 TI - Ethnicity in stroke: practical implications. PMID- 11062304 TI - Recommendations for the management of patients with unruptured intracranial aneurysms: A Statement for healthcare professionals from the Stroke Council of the American Heart Association. PMID- 11062305 TI - AHA Dietary Guidelines: revision 2000: A statement for healthcare professionals from the Nutrition Committee of the American Heart Association. PMID- 11062306 TI - A homozygous deletion in RPE65 in a small Sardinian family with autosomal recessive retinal dystrophy. AB - PURPOSE: We have been engaged in an ongoing study to screen candidate genes for mutations in small families with various forms of autosomal recessive retinal dystrophy. Here we report the screening of a cohort of 14 families from Sardinia for mutations in the genes encoding the alpha- and beta-subunits of cGMP phosphodiesterase and RPE65 (PDE6A, PDE6B, and RPE65). METHODS: Haplotype analysis was performed on each family using simple sequence repeat markers closely flanking or within each of the three gene candidates. For families in which a gene could not be ruled out from segregating with disease, exons of the gene from proband DNAs were screened for mutations by SSCPE (single strand conformation polymorphism electrophoresis). All variants found by SSCPE were sequenced directly. RESULTS: By haplotype analysis, 6/14, 11/14, and 4/13 families were ruled out for PDE6A, PDE6B, and RPE65, respectively. A few variants were found in the proband DNAs of the remaining families, but only one was significant: a 20 bp deletion in exon 4 of RPE65. The deletion co-segregated with disease in one family and caused a frame shift that produces a stop codon downstream. It was absent from the other Sardinian families that we tested, and from Sardinian and North American controls. Results of studies of phenotype in homozygotes and heterozygotes in this Sardinian family are compared with those from a non-Sardinian family recently reported to have the same RPE65 mutation. CONCLUSIONS: This RPE65 mutation, which appears to be quite restricted in its occurrence in Sardinia, leads to childhood onset severe retinal dystrophy or Leber congenital amaurosis. Affecteds of the other 13 plus two additional families were diagnosed with arRP. This family lived in an area of Sardinia where none of the others lived suggesting different ancestral origins. PMID- 11062308 TI - Genetic Polymorphism of Methylenetetrahydrofolate Reductase (MTHFR) and Coronary Artery Disease. AB - A high plasma homocysteine concentration is a risk factor for atherosclerotic disease and venous thrombosis. Homocysteine levels are influenced by folic acid, vitamin B 6 and vitamin B 12, as well as by hereditary factors. A common genetic variant of the methylenetetrahydrofolate reductase (MTHFR) gene CC 677 T) is associated with thermolability of the MTHFR enzyme and elevated plasma homocysteine concentration, especially in those with low folic acid concentration. The prevalence of point mutation (nucleotide 677 C --> T) in MTHFR was measured in patients with coronary artery disease (CAD) who all underwent coronary artery bypass surgery (62 cases; age 64.0 +/- 9.5 years), and was compared with, age-matched control subjects. In patients with coronary artery disease (CAD), we investigated the prevalence of point mutation (nucleotide 677 C --> T) in MTHFR in comparison with control subjects. Heterozygous (C/T) prevalence for the 677 C --> T mutation in the MTHFR was higher in patients with CAD than in control subjects (P < 0.05). The prevalence of homozygosity (C/C) for wild-type MTHFR was lower in patients with CAD in comparison with control subjects (P < 0.05). PMID- 11062307 TI - Further genetic analysis of two autosomal dominant mouse eye defects, Ccw and Pax6(coop). AB - PURPOSE: The work forms part of a major project to study the genetics of mouse cataract mutants found during the course of mutagenesis experiments. The long term aim is to find the underlying gene mutation in each cataract mutant. Here we report further studies of the mutant cataract and curly whiskers (Ccw), previously mapped to Chromosome 4, and also investigations of the corneal opacity (Coop) mutant, which is shown to involve a mutation in the Pax6 gene. METHODS: For Ccw, the methods included mapping relative to microsatellite markers and histological studies. For the Coop mutant, breeding methods were used to show that Coop was allelic with Pax6. The Pax6 coding region in the mutant was then sequenced. RESULTS: The Ccw locus was mapped to approximately position 45cM on the consensus map of Chr 4. Histologically, progressive degeneration of the lens was seen. In the Coop mutant, a base-pair change C->T was found at position 1033 in the Pax6 gene, which created a stop codon leading to premature termination of translation, and to a truncated Pax6 protein. CONCLUSIONS: The phenotype in Ccw/+ heterozygotes involves a new type of lens degeneration in the mouse. On the basis of the phenotype and the locus position, no candidate gene has yet been identified. The Pax6coop mutant differs in phenotype from known null alleles of Pax6, implying that it is a hypomorph. PMID- 11062309 TI - Predictive Factors for the Removal of "Soft" Femoropopliteal Occlusions with Intraarterial Short-term and High-Dose Urokinase Treatment. AB - To date, intraarterial lysis is a widely accepted form of treatment for femoropopliteal occlusions. However, the outcome of therapy has been difficult to predict. Therefore, the present study dealt with two parameters likely to influence the occlusion removal rate. These are the outflow condition distal of the occlusion, and the occlusion length. Fifty patients with subacute femoropopliteal occlusions were treated with intraarterial thrombolysis and, in case residual obstructions were left, with subsequent catheter dilation. Only "soft" occlusions that were probed by a guide wire and proved to be penetrable were chosen for treatment. The occlusion age was 3.79 +/- 4.57 weeks with 90% of cases in the range of 1 day to 6 weeks. Fibrinolytic therapy consisted of one to three one-hour series of 300,000 IU urokinase instillation directly into the occlusion material. The complete clearance rate was 34/50 = 68%. In successfully treated patients, both the pressure gradient (difference between systemic and ankle pressure) and the walking distance improved significantly. Most important factors for success or failure of treatment were the quality of calf outflow and the length of obstruction. Thus, patients with at least one calf artery open and an occlusion length of under 10 cm presented with an opening rate of 88.5%, whereas patients with no calf arteries patent and an occlusion length over 20 cm presented with an opening rate of only 20%. The age of the occlusions, all penetrable by a guide wire prior to treatment, did not influence the rate of recanalization. Intraarterial short-term and high-dose urokinase infusion is an excellent form of treatment for subacute femoropopliteal occlusions. Especially good results with a 88.5% clearance rate were achieved in patients who presented with at least one calf artery open and an occlusion length below 10 cm. PMID- 11062310 TI - Cost-effectiveness of Percutaneous Transluminal Angioplasty (PTA) Versus Vascular Surgery in Limb-threatening Ischaemia. AB - The purpose of this study was to analyze the cost and cost-effectiveness of femoropopliteal PTA compared to femoropopliteal bypass surgery in chronic critical ischaemia of the lower limb. A total of 772 patients were treated either by femoropopliteal PTA or vascular reconstruction in two of the three largest vascular centers in Finland 1991-1992. A subset of 124 cases with chronic critical leg ischaemia, which according to a retrospective independent analysis by a vascular surgeon and a radiologist could have been treated with either modality, were included in the study. Eighty-six of those were treated with PTA and 38 with surgery. The patients were followed up for to three years after treatment. Clinical outcomes were measured as change in the ABI (ankle-brachial pressure index) and avoidance of reoperation and amputation. The hospital costs covering all events from preoperative examinations to the three-year follow-up visit were identified by using hospital discharge register and accounting data. Cost-effectiveness was calculated as cost per reoperation-free year and year of leg saved. Surgery cases were found to have a more severe disease as indicated by lower distal pressures and longer occlusions and they also showed a slightly better clinical outcome, although the differences were not statistically significant. PTA costs were half of those of vascular surgery. The cost effectiveness rates were significantly better for the PTA patients. PTA is a feasible and cost-effective procedure in chronic critical ischaemia of the lower limb and should be the treatment of choice in the subset of patients where both procedures are possible. PMID- 11062311 TI - Antioxidant Activity of Secoisolariciresinol Diglucoside-derived Metabolites, Secoisolariciresinol, Enterodiol, and Enterolactone. AB - Secoisolariciresinol diglucoside (SDG), an antioxidant isolated from flaxseed, is metabolized to secoisolariciresinol (SECO), enterodiol (ED), and enterolactone (EL) in the body. The effectiveness of SDG in hypercholesterolemic atherosclerosis, diabetes, and endotoxic shock could be due to these metabolites. These metabolites may have antioxidant activity. However, the antioxidant activity of these metabolites is not known. The antioxidant activity of SECO, ED, and EL was investigated using chemiluminescence (CL) of zymosan-activated polymorphonuclear leukocytes (PMNLs) [PMNL-CL]. Other antioxidants (SDG and vitamin E) were also used for comparison. SDG, SECO, ED, EL, and vitamin E, each in the concentration of 0.5, 1.0, 2.5, 5.0 and 10.0 mg/ml, produced a concentration-dependent reduction in zymosan-activated PMNL-CL. SDG, SECO, ED, EL, and vitamin E, in the concentration of 2.5 mg/ml, produced a reduction of zymosan-activated PMNL-CL by 23.8%, 91.2%, 94.2%, 81.6% and 18.7%, respectively. Activated PMNLs produce reactive oxygen species and luminol-dependent CL reflects the amount of oxygen species generated from activated PMNLs. The reduction of PMNL-CL, therefore, reflects the antioxidant activity of the compounds studied. These results suggest that the metabolites of SDG have antioxidant activity. The antioxidant activity was highest with SECO and ED and lowest with vitamin E. The antioxidant potency of SECO, ED, EL, and SDG was 4.86, 5.02, 4.35, and 1.27 respectively, as compared to vitamin E. SECO, ED and EL are respectively 3.82, 3.95, and 3.43 more potent than SDG. PMID- 11062312 TI - Comparison of Two Different Arteriovenous Anastomotic Forms By Numerical 3D Simulation of Blood Flow. AB - Anastomotic intimal hyperplasia caused by unphysiological hemodynamics is generally accepted as a reason for dialysis access graft occlusion. Optimizing the venous anastomosis can improve the patency rate of arteriovenous grafts. The purpose of this study was to examine, evaluate and characterize the local hemodynamics, and in particular, wall shear stresses in conventional venous end to-side anastomosis and in patch form anastomosis (Venaflotrade mark) by Computational Fluid Dynamics (CFD). The flow simulations were carried out as three-dimensional to extend results of our previous 2D studies. The numerical simulation was done with a finite volume-based algorithm. The anastomotic forms were constructed with usual size and fixed walls. Subdividing the flow domain into multiple control volumes solved the fundamental equations. The boundary conditions were constant for both forms. The velocity profile of the patch form is better than for the conventional form. The region of high static pressure caused on flow stagnation is reduced on the vein floor. The anastomotic wall shear stress is decreased. The results of this study strongly support patch form use to reduce the incidence of intimal hyperplasia and venous anastomotic stenoses. PMID- 11062313 TI - The Use of Vascular Stents in the Treatment of Iliac Artery Occlusion. AB - To evaluate our experience of selective iliac artery stenting for total occlusions, a prospective observational study of 25 patients with an occluded iliac artery was designed to run from January 1996-May 1997. Exclusion criteria were an occlusion extended to the femoral artery, claudication Grade III or IV, according to the standards for reports dealing with lower extremity ischemia, and vascular (bypass) surgery in the past. Complete recanalization and selective stent placement was possible in all patients. No complications occurred. In one patient re-stenosis happened inside the stent after a year. Percutaneous reintervention was performed with success. The mean ankle-brachial pressure increased from 0.46 before the procedure to 0.95 after the procedure. After two years of follow-up, the mean ankle-brachial pressure is 0.93. The clinical stage improved by at least one grade to Grade 0 (Rutherford classification). The overall probability of patency for occluded iliac arteries in this study was 95% after two years. Recanalization, followed by percutaneous transluminal angioplasty (PTA) in the treatment of iliac artery occlusions, is our first choice of intervention, considering the absence of complication and satisfactory patency rates in patients with claudication Grade I or II. PMID- 11062314 TI - Cytokine Suppressive Agent Prevents Pancreatic Injuries Induced by Ischemia Reperfusion in Rats. AB - This study was designed to evaluate the possible role of cytokines (IL-1 and TNF alpha) in the pathogenesis of pancreatic injuries induced by pancreatic ischemia reperfusion and to evaluate the protective effect of the cytokine suppressive agent, FR167653, against pancreatic injuries. Pancreatic ischemia-reperfusion was induced in rats by ligating the celiac and caudate mesenteric arteries by small metallic clips for 45 min, after this ischemia, the metal clips were removed. Four hours after removing the metal clips, the animals were used for the experiments. In this model, mild hyperamylsemia and significant increases in pancreatic water and trypsin content were observed. Significant increases in serum IL-1 and TNF-alpha were also observed, as compared with the control rats. Pancreatic subcellular redistribution of lysosomal enzyme cathepsin B from the lysosomal fraction to the zymogen fraction was also observed. However, treatment with FR167653 at a dose of 0.5 mg/kg.hr significantly prevented all these pancreatic injuries. These results indicate that cytokines such as IL-1 and TNF alpha might be involved in the pathogenesis of pancreatic injuries induced by ischemia-reperfusion, and that a cytokine suppressive agent might be of therapeutic value for the treatment of pancreatic ischemia. PMID- 11062315 TI - Posterior Wall Reinforcement in Selected Cases of Graft to Aorta End-to-Side Anastomosis. PMID- 11062316 TI - Surgical Management of Giant Descending Aortic Thrombus Detected by Transesophageal Echocardiography. AB - With the advent of transesophageal echocardiography (TEE), giant thrombi of the descending aorta are becoming increasingly recognized as possible sources of peripheral emboli. This report describes the management of three patients presenting with multiple unexplained peripheral emboli. All three patients were treated successfully with aortic thrombectomy and long-term anticoagulation. PMID- 11062317 TI - Ultrasound Estimates of Venous Valve Function in Screening For Insufficiency and Following Patients With Chronic Venous Disease. PMID- 11062318 TI - Unusual Case of Klippel-Trenaunay Syndrome A Case Report and Review of the Literature. AB - A 15-year-old Indian boy from Guyana presented with episodes of syncope for past 4 years. Klippel-Trenaunay was entertained based on presence of capillary hemangioma and gross venous malformation of left leg. Unusual features noticed were "orthostatic hypotension as a presentation," atrophy of leg as contrasted to common finding of hypertrophy, skeletal hypoplasia, abnormal venous and arterial features. Surgical management was undertaken in view of large venous pooling, and abnormal veins threatening to rupture, with good results. PMID- 11062319 TI - 42(nd) Annual World Congress International College of AngiologySan Diego, California * 25-30 June 2000Abstracts and Posters Presented. PMID- 11062320 TI - Developmental changes within the core of artifact concepts. AB - Three experiments addressed the relative importance of original function and current function in artifact categorization. Subjects were asked to judge whether an artifact that was made for one purpose (e.g. making tea) and was currently being used for another purpose (e.g. watering flowers) was a teapot or a watering can. Experiment 1 replicated the finding by Hall (1995) (unpublished manuscript) that adults rely on the original function of an artifact over a current function in their kind judgments. Experiments 2 and 3 revealed that whereas the kind judgments of 6-year-olds, like those of adults, patterned with the original function, those of 4-year-olds did not. Four-year-olds were influenced by the order in which the functions were mentioned in the story. Further, in their justifications 6-year-olds and adults referred to the origin of the objects, whereas 4-year-olds virtually never did. We conclude that 6-year-olds have begun to organize their understanding of artifacts around the notion of original function, and that 4-year-olds have not. The data are discussed as they bear on children's understanding of the design stance (Dennett, D. C. (1987). PMID- 11062321 TI - Altering object representations through category learning. AB - Previous research has shown that objects that are grouped together in the same category become more similar to each other and that objects that are grouped in different categories become increasingly dissimilar, as measured by similarity ratings and psychophysical discriminations. These findings are consistent with two theories of the influence of concept learning on similarity. By a Strategic Judgment Bias account, the categories associated with objects are explicitly used as cues for determining similarity, and objects that are categorized together are judged to be more similar because similarity is not only a function of the objects themselves, but also the objects' category labels. By a Changed Object Description account, category learning alters the description of the objects themselves, emphasizing properties that are relevant for categorization. A new method for distinguishing between these accounts is introduced which measures the difference between the similarity ratings of categorized objects to a neutral object. The results indicate both strategic biases based on category labels and genuine representational change, with the strategic bias affecting mostly objects belonging to different categories and the representational change affecting mostly objects belonging to the same category. PMID- 11062322 TI - Language and number: a bilingual training study. AB - Three experiments investigated the role of a specific language in human representations of number. Russian-English bilingual college students were taught new numerical operations (Experiment 1), new arithmetic equations (Experiments 1 and 2), or new geographical or historical facts involving numerical or non numerical information (Experiment 3). After learning a set of items in each of their two languages, subjects were tested for knowledge of those items, and new items, in both languages. In all the studies, subjects retrieved information about exact numbers more effectively in the language of training, and they solved trained problems more effectively than untrained problems. In contrast, subjects retrieved information about approximate numbers and non-numerical facts with equal efficiency in their two languages, and their training on approximate number facts generalized to new facts of the same type. These findings suggest that a specific, natural language contributes to the representation of large, exact numbers but not to the approximate number representations that humans share with other mammals. Language appears to play a role in learning about exact numbers in a variety of contexts, a finding with implications for practice in bilingual education. The findings prompt more general speculations about the role of language in the development of specifically human cognitive abilities. PMID- 11062323 TI - Are there principles that apply only to the acquisition of words? A reply to Waxman and Booth. PMID- 11062324 TI - Recognizing one's own face. AB - We report two studies of facial self-perception using individually tailored, standardized facial photographs of a group of volunteers and their partners. A computerized morphing procedure was used to merge each target face with an unknown control face. In the first set of experiments, a discrimination task revealed a delayed response time for the more extensively morphed self-face stimuli. In a second set of experiments, functional magnetic resonance imaging (fMRI) was used to measure brain activation while subjects viewed morphed versions of either their own or their partner's face, alternating in blocks with presentation of an unknown face. When subjects viewed themselves (minus activation for viewing an unknown face), increased blood oxygenation was detected in right limbic (hippocampal formation, insula, anterior cingulate), left prefrontal cortex and superior temporal cortex. In the partner (versus unknown) experiment, only the right insula was activated. We suggest that a neural network involving the right hemisphere in conjunction with left-sided associative and executive regions underlies the process of visual self-recognition. Together, this combination produces the unique experience of self-awareness. PMID- 11062325 TI - Reading time evidence for enriched composition. AB - Verbs like 'begin' and 'enjoy' appear to semantically select a complement that expresses an activity or an event (Jackendoff, R. (1997). The architecture of the language faculty. Cambridge, MA: MIT Press; Pustejovsky, J. Cognition 41 (1991) 47; Pustejovsky, J. (1995). The generative lexicon. Cambridge, MA: MIT Press). When these verbs occur with a complement that does not directly denote either an activity or an event (e.g. '... began the book' or '... enjoyed the book'), the complement must be type-shifted from an object to an activity to conform to the verb's semantic restrictions. We report an experiment in which self-paced reading times were found to be longer for complements that required type-shifting than for complements that directly matched the semantic restrictions of the matrix verb. This finding is argued to provide on-line evidence for a type of enriched lexical processing posited in recent lexical semantic research. PMID- 11062326 TI - Angiotensin II mediated signal transduction. Important role of tyrosine kinases. AB - It has been 100 years since the discovery of renin by Bergman and Tigerstedt. Since then, numerous studies have advanced our understanding of the renin angiotensin system. A remarkable aspect was the discovery that angiotensin II (AngII) is the central product of the renin-angiotensin system and that this octapeptide induces multiple physiological responses in different cell types. In addition to its well known vasoconstrictive effects, growing evidence supports the notion that AngII may play a central role not only in hypertension, but also in cardiovascular and renal diseases. Binding of AngII to the seven-transmembrane angiotensin II type 1 receptor is responsible for nearly all of the physiological actions of AngII. Recent studies underscore the new concept that activation of intracellular second messengers by AngII requires tyrosine phosphorylation. An increasing number of tyrosine kinases have been shown to be activated by AngII, including the Src kinase family, the focal adhesion kinase family, the Janus kinases and receptor tyrosine kinases. These actions of AngII contribute to the pathophysiology of cardiac hypertrophy and remodeling, vascular thickening, heart failure and atherosclerosis. In this review, we discuss the important role of tyrosine kinases in AngII-mediated signal transduction. Understanding the importance of tyrosine phosphorylation in AngII-stimulated signaling events may contribute to new therapies for cardiovascular and renal diseases. PMID- 11062327 TI - Interaction of the catecholamine release-inhibitory peptide catestatin (human chromogranin A(352-372)) with the chromaffin cell surface and Torpedo electroplax: implications for nicotinic cholinergic antagonism. AB - The catecholamine release-inhibitory chromogranin A fragment catestatin (chromogranin A(344-364)) exhibits non-competitive antagonism of nicotinic cholinergic signaling in chromaffin cells. A previous homology model of catestatin's likely structure suggested a mode of interaction of the peptide with the nicotinic receptor, but direct evidence has been lacking. Here we found that [125I]-catestatin binds to the surface of intact PC12 and bovine chromaffin cells with high affinity (K(D)=15.2+/-1.53 nM) and specificity (lack of displacement by another [N-terminal] fragment of chromogranin A). Nicotinic agonist (carbamylcholine) did not displace [125I]-catestatin from chromaffin cells, nor did catestatin displace the nicotinic agonist [3H]-epibatidine; these observations indicate a catestatin binding site separate from the agonist binding pocket on the nicotinic receptor, a finding consistent with catestatin's non competitive nicotinic mechanism. [125I]-catestatin could be displaced from chromaffin cells by substance P (IC(50) approximately 5 microM), though at far lower potency than displacement by catestatin itself (IC(50) approximately 350 380 nM), suggesting that catestatin and substance P occupy an identical or overlapping non-competitive site on the nicotinic receptor, at different affinities (catestatin > substance P). Small, non-peptide non-competitive nicotinic antagonists (hexamethonium or clonidine) did not diminish [125I] catestatin binding, suggesting distinct non-competitive binding sites on the nicotinic receptor for peptide and non-peptide antagonists. Similar binding and inhibitory profiles for [125I]-catestatin were observed on chromaffin cells as well as nicotinic receptor-enriched Torpedo membranes. Covalent cross-linking of [125I]-catestatin to Torpedo membranes suggested specific contacts of [125I] catestatin with the delta, gamma, and beta subunits of the nicotinic receptor, a finding consistent with prior homology modeling of the interaction of catestatin with the extracellular face of the nicotinic heteropentamer. We conclude that catestatin occludes the nicotinic cation pore by interacting with multiple nicotinic subunits at the pore vestibule. Such binding provides a physical explanation for non-competitive antagonism of the peptide at the nicotinic receptor. PMID- 11062328 TI - Effects of galanin on wide-dynamic range neuron activity in the spinal dorsal horn of rats with sciatic nerve ligation. AB - Galanin is a 29-amino acid peptide with a suggested role in nociception. The effect of galanin on wide-dynamic range neuron discharge frequency in rats with nerve ligation, used as a model of neurogenic pain, was investigated by extracellular recording methods. Seven to 14 days after sciatic nerve ligation, 0.1, 0.5 or 1 nmol of galanin was administered directly on the dorsal surface of the L3-L5 spinal cord of rats with sciatic nerve ligation. It was found that galanin inhibited the activity of wide-dynamic range neurons dose-dependently, an effect was more pronounced in sciatic nerve ligated rats than intact rats. Furthermore, when 1 nmol of galantide, the galanin antagonist, was administered on the dorsal surface of the L3-L5 spinal cord, the wide-dynamic range neuron discharge frequency increased significantly. The results suggest that galanin plays an important role in the modulation of presumed nociception in mononeuropathy. PMID- 11062329 TI - Molecular evolution of the neuropeptide Y (NPY) family of peptides: cloning of three NPY-related peptides from the sea bass (Dicentrarchus labrax). AB - Neuropeptide Y (NPY) is a 36-amino-acid peptide that is widely and abundantly expressed in the central nervous system of all vertebrates investigated. Related peptides have been found in various vertebrate groups: peptide YY (PYY) is present in gut endocrine cells of many species and pancreatic polypeptide (PP) is made in the pancreas of all tetrapods. In addition, a fish pancreatic peptide called PY has been reported in three species of fishes. The evolutionary relationships of fish PY have been unclear and it has been proposed to be the orthologue (species homologue) of each of the three tetrapod peptides. We demonstrate here with molecular cloning techniques that the sea bass (Dicentrarchus labrax), an acanthomorph fish, has orthologues of both NPY and PYY as well as a separate PY peptide. Sequence comparisons suggest that PY arose as a copy of the PYY gene, presumably in a duplication event separate from the one that generated PP from PYY in tetrapods. PY sequences from four species of fish indicate that, similar to PP, PY evolves much more rapidly than NPY and PYY. The physiological role of PY is unknown, but we demonstrate here that sea bass PY, like NPY and PYY but in contrast to the tetrapod PP, is expressed in brain. PMID- 11062330 TI - The NPY effects on murine leukocyte adherence and chemotaxis change with age. Adherent cell implication. AB - The two-way communication between the nervous and immune system is currently well known, but the age-related changes in this communication have been scarcely studied. In the present work, we have investigated the in vitro effects of neuropeptide Y (NPY) at concentrations ranging from 10(-13) to 10(-7) M on the adherence and chemotaxis capacities of spleen, axillary node, thymus and peritoneum leukocytes from BALB/c mice. The NPY effect on these functions was examined on cells from animals of four different ages, i.e. young (12+/-2 weeks old), adult (24+/-2 weeks old), mature (50+/-2 weeks old) and old (72+/-2 weeks old). In young animals, NPY stimulates the adherence of leukocytes from spleen, axillary nodes and thymus and inhibits it in cells from peritoneum. In adult animals NPY inhibits the adherence of leukocytes from thymus. These effects disappear with ageing in all locations. Chemotaxis is stimulated by this neuropeptide at all ages in cells from axillary nodes and peritoneum, but this effect is absent in old mice. NPY exerts an inhibitory effect on the chemotaxis of leukocytes from thymus at all ages studied. These NPY effects on leukocytes seem to be carried out through adherent cells. PMID- 11062331 TI - Acceleration of pubertal development following central blockade of the Y1 subtype of neuropeptide Y receptors. AB - Pubertal development results from the coordinate secretion of gonadotropin releasing hormone (GnRH) by hypothalamic GnRH neurons. Central administration of neuropeptide Y (NPY) to prepubertal rats can indefinitely delay sexual maturation by inhibiting this GnRH secretion. The aim of the present study was to further investigate the physiological role of NPY in pubertal development, and to assess the potential involvement of its Y1 receptor subtype in this setting. The timing of pubertal development was determined in juvenile female rats receiving chronic i.c.v. infusion of a specific Y1 receptor antagonist (BIBP 3226), and compared with controls. Although treatment with BIBP 3226 did not affect the age at vaginal opening, animals receiving the Y1 antagonist experienced a quicker progression through puberty, corroborated by a significant increase in pituitary luteinizing hormone content. This effect of BIBP3226 on the gonadotrope axis occurred without apparent toxicity, but was accompanied by a transient decrease in body weight gain on the first day of treatment, suggesting an effect on appetite. Together, our results add to the evidence in favour of a role for NPY in the onset of puberty. They are entirely consistent with the proposed inhibition exerted by endogenous hypothalamic NPY before the onset of pubertal development. They also suggest that the Y1 subtype of NPY receptors is involved in this effect. PMID- 11062332 TI - Multiple regulation of adenylyl cyclase activity by G-protein coupled receptors in human foetal lung fibroblasts. AB - The pharmacological profile of adenylyl cyclase activity was analysed in WI-38 human foetal lung fibroblasts. Among various agents that act through G-protein coupled receptors, only the beta-adrenergic agonist isoproterenol stimulated and the tetradecapeptide somatostatin (SRIF, sst) inhibited the enzyme activity. The use of the reverse transcription-polymerase chain reaction (RT-PCR) methodology with appropriate cDNAs allowed us to identify the expression of four subtypes of SRIF transmembrane receptors (sst1-4 but not sst5 receptors) in this cell line. By RT-PCR and immunochemistry techniques, we also demonstrated the expression of stimulatory (alpha(s)) and inhibitory (alpha(i1), alpha(i2) and alpha(i3)) G protein subunits. The known role of the adenylyl cyclase system in cell proliferation and differentiation mechanisms together with the present analysis of the corresponding regulatory network in fibroblasts of human foetal lung add knowledge on the cell line WI-38 that is widely used as a model system in studying cell growth. The importance of this cell class in normal and abnormal lung function and development reinforces the significance of these results. PMID- 11062333 TI - Neuropeptide Y Y1 receptor mediated mesenteric vasoconstriction in the pig in vivo. AB - The object of the present study was to investigate the effects of the sympathetic cotransmitter neuropeptide Y (NPY), and the closely related gut hormone peptide YY (PYY), on splanchnic blood flow regulation in the anaesthetized pig in vivo. Systemic injections of NPY, PYY and the NPY Y(1) receptor agonist [Leu(31)Pro(34)]NPY (470 pmol kg(-1) each) evoked pressor and mesenteric vasoconstrictor responses that were largely abolished by the selective NPY Y(1) receptor antagonist H 409/22 (60 nmol kg(-1) min(-1)). In contrast, the NPY Y(2) receptor agonist N-acetyl[Leu(28)Leu(31)]NPY(24-36) (1.1 nmol kg(-1)), a dose of which potently evoked splenic NPY Y(2) receptor mediated (not affected by H 409/22) vasoconstriction, did not evoke any mesenteric vascular response. Mesenteric vascular responses to angiotensin II (10 pmol kg(-1)), alpha,beta methylene ATP (10 nmol kg(-1)) and the alpha(1)-adrenoceptor agonist phenylephrine (15 nmol kg(-1)), were not inhibited by H 409/22. It is concluded that NPY and PYY evokes porcine mesenteric vasoconstriction mediated by the NPY Y(1) receptor subtype, as demonstrated by selective and specific inhibition exerted by the NPY Y(1) receptor antagonist H 409/22, in vivo. PMID- 11062334 TI - Guanylyl cyclase-C receptor mRNA distribution along the rat nephron. AB - Guanylin (GN) and uroguanylin (UGN) are two recently identified peptides that have been shown to affect water and electrolyte transport in both the intestine and the kidney. Mechanistically, the effects of both peptides are thought to be mediated by intracellular cGMP which results from ligand binding to a plasma membrane guanylyl cyclase-C (GC-C) receptor. To date, the specific intrarenal site(s) of GN and UGN action have not been established. To begin to address this issue, the present studies utilized semi-quantitative RT-PCR to assess the distribution of GC-C mRNA in specific microdissected segments of the rat nephron. GC-C mRNA expression was highest in the cortical collecting tubule, followed by the proximal convoluted tubule, medullary thick ascending limb and collecting tubule, and thin limbs of Henle's loop. Expression levels were significantly lower in all other segments tested, including the glomerulus. The renal tubular expression pattern for cGMP-dependent protein kinase II (cGK-II) mRNA, which is activated in response to GN/UGN-dependent cGMP accumulation, was similar to that for GC-C. Notably, both GN and UGN mRNAs were also expressed along the nephron. The highest levels of expression for both peptides were detected in the medullary collecting tubule. Lower, but comparable levels of GN and UGN expression also occurred in the cortical collecting tubule, cortical and medullary thick ascending limb, and thin limbs of Henles loop. In the proximal convoluted tubule, GN mRNA expression was also quite high, while UGN mRNA was almost undetectable. The presence of renal GC-C and cGK-II in the kidney are consistent with a proposed endocrine function for GN and UGN. In addition however, the present data suggest that intrarenally synthesized GN and UGN may also contribute to the regulation of renal tubular transport. PMID- 11062335 TI - Inhibitory neurogenic modulation of histamine-induced cutaneous plasma extravasation in the pigeon. AB - The neurohumoral modulation of the permeability increasing effect of histamine was studied in pigeon skin. Substances were administered through plasmapheresis capillaries inserted into the dorsal wing skin and the protein contents of the perfusates were determined by a quantitative method. The vascular labelling technique was also utilized to histologically identify leaky blood vessels. In the innervated skin histamine evoked a significant, dose-dependent plasma extravasation which was markedly augmented by the coadministration of a specific galanin receptor antagonist, galanin-1-16-bradykinin-2-9-amide (M35). Chronic cutaneous denervation per se resulted in a significant elevation of the permeability-enhancing effect of histamine. In the denervated skin this response was not affected by M35 but was significantly inhibited by galanin. It is concluded that in the normally innervated skin endogenous galanin may exert a neurogenic tonic inhibitory effect on histamine-induced plasma leakage. It is suggested that sensory nerves possess not only pro-inflammatory, but also anti inflammatory (inhibitory) sensory-efferent functions. PMID- 11062336 TI - Organ-specific mRNA distribution of C-type natriuretic peptide in neonatal and adult mice. AB - C-type natriuretic peptide (CNP) is described as an endothelium-derived vasodilator and a growth inhibitor of vascular smooth muscle cells. In the present study, CNP mRNA was quantified by RNase-protection assay to elucidate organ distribution of CNP in neonatal and adult mice. In adult mice, the highest CNP expressions were detected in uterus and ovary, which exceeded the CNP concentrations of forebrain and brainstem. In contrast, neonatal mice showed highest CNP-mRNA levels in forebrain and brainstem with lower levels in skin, tongue, heart, lung, thymus, skeletal muscle, liver, kidney, stomach, and skull. Thus, CNP-expression pattern diminishes during postnatal development. The observation that the expression level of CNP mRNA is 2.2-fold higher in the adult forebrain compared to the neonatal forebrain allows a comparison between all neonatal and adult organs. PMID- 11062337 TI - Recombinant granulocyte colony-stimulating factor induces production of human neutrophil peptides in lung cancer patients with neutropenia. AB - Human neutrophil peptides (HNPs) 1, 2 and 3 are antimicrobial peptides localized in the azurophil granules of neutrophils. We investigated the effects of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on the biosynthesis of HNPs 1-3 using a sensitive radioimmunoassay and Northern blot analysis. Seven patients with lung cancer were first treated with various anticancer agents for 3 days (days 1-3) followed by treatment with rhG-CSF (2 microgram/kg weight/day) for 7 days (days 8-14). Chemotherapy caused neutropenia but the neutrophil count increased biphasically between days 8 and 14. Chemotherapy did not change the baseline plasma concentration of HNPs 1-3 (74.1+/ 2.1 pmol/ml) but the concentration increased from day 12, 5 days after commencement of rhG-CSF therapy, to reach a peak value of 430.8+/-57.0 pmol/ml on day 15, 1 day after the last administration of rhG-CSF. Baseline HNPs 1-3 content per neutrophil was 0.59+/-0.02 fmol, decreased to 0.30+/-0.07 fmol on day 9, then increased to 0.78+/-0.07 fmol on day 15. Analyses of peripheral blood neutrophils by Northern blot and reverse-phase high-performance liquid chromatography showed that the amounts of HNPs 1-3 mRNA and precursors of HNPs 1-3 markedly increased in response to rhG-CSF. Our results indicate that recombinant hG-CSF does not only increase neutrophil count but stimulates HNPs 1-3 biosynthesis in neutrophils, thus enhancing the host defense system of compromised hosts with neutropenia. PMID- 11062338 TI - Effects of des-aspartate-angiotensin I on angiotensin II-induced incorporation of phenylalanine and thymidine in cultured rat cardiomyocytes and aortic smooth muscle cells. AB - Des-aspartate-angiotensin I, a pharmacologically active nine-amino acid angiotensin peptide, and losartan, an AT(1) angiotensin receptor antagonist, but not angiotensin-(1-7), another active angiotensin peptide, completely attenuated the angiotensin II-induced incorporation of [3H]phenylalanine in cultured rat cardiomyocytes. The attenuation by des-aspartate-angiotensin I but not that of losartan was inhibited by indomethacin. The data support an earlier suggestion that the nonapeptide attenuates cardiac hypertrophy in rats via an indomethacin sensitive angiotensin AT(1) receptor subtype. In rat aortic smooth muscle cells, both des-aspartate-angiotensin I and angiotensin-(1-7) had no effect on the angiotensin II-induced [3H]phenylalanine incorporation. However, the two peptides significantly attenuated the angiotensin II-induced [3H]thymidine incorporation in the smooth muscle cells. The attenuation by angiotensin-(1-7) but not by des aspartate-angiotensin I was inhibited by (D-Ala(7))-angiotensin-(1-7), a specific angiotensin-(1-7) antagonist. Des-aspartate-angiotensin I also attenuated FCS stimulated [3H]thymidine incorporation. This attenuation was inhibited by the peptide angiotensin receptor antagonist, (Sar(1), Ile(8))-angiotensin II, but not by losartan. These data indicate that des-aspartate-angiotensin I and angiotensin (1-7) do not participate in the process of protein synthesis in vascular smooth muscle cells and that the nonapeptide and heptapeptide act on different non-AT(1) receptors to mediate their anti-hyperplasic action. Although the exact mechanisms of action remain to be elucidated, the findings indicate that des-aspartate angiotensin I acts as an agonist on angiotensin AT(1) and non-AT(1) receptor subtypes and induces responses that oppose the actions of angiotensin II. PMID- 11062339 TI - Angiotensin-(1-7) regulates the levels of angiotensin II receptor subtype AT1 mRNA differentially in a strain-specific fashion. AB - Ang-(1-7) is an effector peptide of the renin-angiotensin system with several distinct actions that are likely mediated by a specific receptor. Regulatory effects of angiotensin (Ang) peptides, Ang-(1-7) and Ang II, on Ang receptor subtype 1 (AT1) mRNA expression were investigated in vascular smooth muscle cells (VSMC) from four University of Akron (Akr) rat strains (WKY, SHR and two backcross consomic lines SHR/y and SHR/a), and in SHR and WKY cells from Charles River Laboratories (Crl). In WKY/Akr and SHR/Akr, Ang-(1-7) treatment increased the levels of AT1 mRNA. This effect was inhibited by the specific Ang-(1-7) antagonist, A-779, in WKY/Akr but not SHR/Akr. Ang II had no effect in Akr cells, but it down-regulated AT1 mRNA in WKY/Crl and SHR/Crl VSMC. Ang-(1-7) did not affect AT1 mRNA levels in Crl lines. In conclusion, Ang-(1-7) regulates the AT1 receptor either directly or indirectly in a strain-specific fashion. The Ang-(1 7) antagonist, A-779, blocks the actions of Ang-(1-7) only in VSMC from WKY/Akr rats, suggesting either that the binding sites for Ang-(1-7) have different properties in SHR/Akr and WKY/Akr cell lines, or that some of the effects of Ang (1-7) are not receptor mediated. Further, we found differences between Akr cells and Crl cells that are consistent with their genetic heterogeneity. PMID- 11062340 TI - Involvement of neuropeptide Y and Y1 receptor in antinociception in nucleus raphe magnus of rats. AB - The nociceptive response latencies increased significantly after intra-nucleus raphe magnus administration of 0.1 or 0.4 nmol of neuropeptide Y, but not 0.04 nmol, in rats. The neuropeptide Y-induced increases in hindpaw withdrawal latency were reversed by following injection of 0.42 nmol of the Y1 antagonist, NPY(28 36). The results indicate that NPY plays an antinociceptive role in nucleus raphe magnus in rats, which is mediated by the Y1 receptor. Furthermore, the neuropeptide Y-induced increases in hindpaw withdrawal latency were attenuated by following intra-nucleus raphe magnus injection of 6 nmol of the opioid antagonist naloxone, indicating that there is an interaction between NPY and opioids in nucleus raphe magnus. PMID- 11062341 TI - Pro-xenopsin(s) in vesicles of mammalian brain, liver, stomach and intestine is apparently released into blood and cerebral spinal fluid. AB - Mammalian pro-xenopsins (proXP), proteins (such as alpha-coatomer) that yield XP related peptides when digested by pepsin-related proteases, are ubiquitously distributed in rats, with highest concentrations in liver and gastrointestinal tissues. Here, the cellular and subcellular distributions of canine and rat proXP were determined in brain, liver, stomach and intestine. Elutriation and percoll density centrifugation of collagenase-dispersed cells demonstrated that proXP was primarily associated with hepatocytes in liver, chief and parietal cells in stomach and endocrine/exocrine cells in intestine. When fragmented cells were subjected to differential centrifugation, congruent with85% of proXP was associated with particulate fractions and only congruent with15% was cytosolic. Sucrose-gradient centrifugation of crude mitochondrial preparations (P2 pellets) for liver, stomach and intestine demonstrated that proXP was localized to vesicles (density, congruent with1.19; size, 80-400 micrometer), which contained material of variable electron density. In isotonic homogenates of brain, proXP migrated primarily with synaptosomes (density, congruent with1. 15) which contained vesicles (size, 50-100 micrometer). During HPLC-sizing and ion exchange chromatography, proXP gave at least three components, the major one being an anionic 140-kDa protein. ProXP-like activity was found in human and rat blood, human cerebral spinal fluid and in contents of the gastrointestinal lumen. These results are consistent with the idea that these vesicle-associated protein(s) could be released during endocrine and/or exocrine secretion and serve as precursors to XP-related peptides. PMID- 11062342 TI - Structure, function, and regulation of tartrate-resistant acid phosphatase. PMID- 11062343 TI - Risk of hip fracture according to the World Health Organization criteria for osteopenia and osteoporosis. AB - The risk of hip fracture is commonly expressed as a relative risk. The aim of this study was to examine the utility of relative risks of hip fracture in men and women using World Health Organization (WHO) diagnostic criteria for low bone mass and osteoporosis. Reference data for bone mineral density (BMD) at the femoral neck, from the third National Health and Nutrition Examination Survey (NHANES III), were applied to the population of Sweden. Relative risks (RRs) were calculated from the known relationship between BMD at the femoral neck and hip fracture risk. The apparent prevalence of low bone mass and osteoporosis depended on the segment of the young population chosen for reference ranges. Using a reference derived from women aged 20-29 years, the prevalence of osteoporosis was 21.2% in women between the ages of 50 and 84 years and 6.3% in men. The RRs associated with osteoporosis depended markedly on the risk comparison. For example, in men or women aged 50 years, the RR of hip fracture in those with osteoporosis compared to those without osteoporosis was 7.4 and 6.1, respectively. The RR of those at the threshold value for osteoporosis compared to those with an average value for BMD at that age was 6.6 and 4.6 in men and women, respectively. RRs were lower comparing those at the threshold value compared to the risk of the general population at that age (4.2 and 2.9, respectively). When RR was expressed in relation to the population risk rather than to the risk at the average value for BMD, RR decreased at all ages by 37%. Such adjustments are required for risk assessment in individuals and for the combined use of different risk factors. Because the average T score at each age decreased with age, the RR of hip fracture at any age decreased with advancing age in the presence of osteoporosis. The decrease in relative risk with age is, however, associated with an increase in absolute risk. Thus, for clinical use, the expression of absolute risks may be preferred to relative risks. PMID- 11062344 TI - Human Dermo-1 has attributes similar to twist in early bone development. AB - Basic helix-loop-helix (bHLH) transcription factors are implicated in cell lineage determination and differentiation. Dermo-1 encodes a bHLH transcription factor that shares extensive homology with another bHLH transcription factor, Twist. We have cloned and characterized human Dermo-1 from two different bone cytoplasmic DNA (cDNA) libraries. Dermo-1 mRNA and protein expression were examined in human embryo and adult tissue sections. Dermo-1 is expressed in a subset of mesodermally and ectodermally derived tissues. We further examined expression of Dermo-1/Twist in human tissues and cell lines. In addition, we observed Dermo-1 expression in response to basic fibroblast growth factor in osteoblastic cell lines. To evaluate the functionality of the human Dermo-1 transcription factor in osteoblast metabolism, we made stable osteoblastic cell lines that over- and underexpress human Dermo-1. These cell lines were analyzed and compared with previously published data of similar cell lines transfected with Twist. Our results demonstrate that Dermo-1 caused changes similar to Twist in the osteogenic properties of osteoblastic cells, such as morphology, bone marker gene expression, and biochemical response to cytokines. However, Dermo-1 expression also has unique effects in regulating the mechanism of proliferation, on alkaline phosphatase enzyme activity, and in temporal expression patterns. We speculate that expression of Twist and Dermo-1 maintains cells in an osteoprogenitor or preosteoblast-like state, respectively, and prevents premature or ectopic osteoblast differentiation. Therefore, Twist and Dermo-1 must be sequentially downregulated in order to initiate the cascade of events responsible for osteogenic cell differentiation. These results indicate that, during osteoblast development, Dermo-1 may inhibit osteoblast maturation and maintain cells in a preosteoblast phenotype by utilizing mechanisms similar but not identical to those utilized by Twist. PMID- 11062345 TI - Progressive increase in bone mass and development of odontomas in aging osteopetrotic c-src-deficient mice. AB - The critical role of c-src in osteoclast-mediated bone resorption has been emphasized by gene deletion experiments in mice. However, the long-term effects of the lack of c-src and impaired osteoclast function on the skeleton remain unknown. To further study the physiological role of c-src and to circumvent the early death of src(-/-) mice, due to starvation in the absence of erupted teeth, we maintained mice on a liquid diet. At the age of 2 months the src(-/-) mice presented signs of airway obstruction and all mice died progressively between 2.5 and 6 months of age. Radiography demonstrated severe osteopetrosis of the whole skeleton. Histomorphometrical analysis of the src(-/-) mice confirmed a significant increase in bone mass with age, resulting in complete loss of bone marrow spaces in some bones and explaining the consistent hepatosplenomegaly, due to extraskeletal hematopoesis. Histopathological examination of the skull revealed the presence of odontomas in the region of the unerupted incisors, with a penetrance of 100% in the aging src(-/-) mice. Although odontomas are benign lesions, their progressive growth leads to the obliteration of the nasal airways, progressive suffocation, and death in src(-/-) mice. These results suggest that: (i) in the absence of bone resorption, bone formation continues and leads to progressive accentuation of the osteopetrotic phenotype in src(-/-) mice; (ii) osteoclastic function is required for regular eruption of the incisors and deficient bone resorption is associated with the development of odontomas; and (iii) src(-/-) mice die by suffocation due to airway obliteration as a result of progressive odontoma growth. PMID- 11062346 TI - Skeletal effects of estrogen deficiency as induced by an aromatase inhibitor in an aged male rat model. AB - Aromatization of androgens into estrogens may be important for maintenance of the male skeleton. To address this hypothesis, we evaluated the skeletal effects of selective estrogen deficiency as induced by the aromatase inhibitor vorozole (Vor), with or without 17beta-estradiol (E(2)) administration (1.35 microg/day), in aged (12-month-old) male rats. A baseline group was killed at the start of the experiment (Base). The control group (Control), the group treated with vorozole alone (Vor), the group treated with E(2) alone (E(2)), or the group with a combination of both (Vor + E(2)) were killed 15 weeks later. Vorozole significantly increased serum testosterone (T) and reduced serum E(2) compared with Control. Body weight gain and serum insulin-like growth factor-I (IGF-I) were also lower in Vor, whereas significant weight loss and decrease of serum IGF I occurred as a result of E(2) administration. Bone formation as assessed by serum osteocalcin was unaffected but osteoid surface in the proximal metaphysis of the tibia was increased in Vor-treated rats. Bone resorption as evaluated by urinary deoxypyridinoline excretion was increased in Vor. Biochemical parameters of bone turnover were reduced significantly in all E(2) treated rats. Premature closure of the growth plates and decreased osteoid and mineralizing surfaces were also observed in E(2) and Vor + E(2). Apparent bone density of lumbar vertebrae and femur, as measured by dual-energy X-ray absorptiometry (DXA), was significantly reduced in Vor. Vorozole decreased femoral bone density mainly in the distal femur (trabecular and cortical region). This decrease of bone density was not present in E(2) and Vor + E(2). Similar findings were observed when bone density was assessed by peripheral quantitative computed tomography (pQCT); that is, trabecular density of the distal femur, the proximal tibia, and the distal lumbar vertebra were all lower in Vor. This decrease in density was not observed in all E(2)-treated animals. In conclusion, administration of the aromatase inhibitor, vorozole, to aged male rats induces net trabecular bone loss in both the appendicular and axial skeleton, despite a concomitant increase in serum testosterone. E(2) administration is able to prevent this trabecular bone loss in vorozole-treated animals. PMID- 11062347 TI - Spontaneous fracture (sfx): a mouse genetic model of defective peripubertal bone formation. AB - A new mouse model of stage-specific bone growth failure and fracture has been recovered as an autosomal recessive mutation, designated spontaneous fracture (sfx). The sfx/sfx mice are phenotypically normal until shortly after weaning, when reduced mobility and impaired somatic growth are first noted. By 6 weeks of age, body, spleen, and thymus weights, as well as hematocrits and serum calcium, inorganic phosphate, total alkaline phosphatase, insulin-like growth factor-I, and osteocalcin levels are decreased. The sfx/sfx mice also show reduced femoral cortical density and diaphyseal circumference, as well as a paucity of mature osteoblasts on bone surfaces. Histological analyses of the femur and tibia in the mutants show subtle reduction of chondrocyte numbers in epiphyseal-plate columns, reduction of matrix, and near absence of osteoid below the differentiated chondrocytes. Trabeculae in proximal tibiae, iliacs, and vertebral bodies are sparse and thin. Cortical bone thickness of mutants is markedly thinned in all sites examined. By 7-8 weeks, radiographic films routinely show spontaneous impact fractures of the distal femur accompanied by callus formation, whereas complete fractures are less commonly observed. Volumetric bone mineral density (BMD) of mutant femurs is similar to +/? littermates in the center of the femoral diaphysis, but BMD declines as either end of the femoral diaphysis is approached. We have mapped the gene responsible for this phenotype to central Chromosome 14. Reduced bone mass, impaired bone formation, abnormalities of bone architecture, and a disposition to spontaneous fracture identify sfx/sfx mice as a useful model for understanding the mechanisms responsible for peripubertal bone formation. PMID- 11062348 TI - Osteoclasts differentiate from resident precursors in an in vivo model of synchronized resorption: a temporal and spatial study in rats. AB - Osteoclasts differentiate from mononucleated precursors expressing monocyte markers, which gradually evolve to preosteoclasts expressing the osteoclast phenotype. Although the role of osteogenic cells in these changes has been well documented in vitro, their contribution in vivo has not been established. In this study, a synchronized wave of resorption was activated along the mandibular periosteum. The periosteum adjacent to the bone surface studied was separated by a computer-assisted technique into an osteogenic alkaline phosphatase-positive compartment and an outer nonosteogenic compartment. Specific markers (nonspecific esterase [NSE], tartrate-resistant acid phosphatase [TRAP], and ED1 antibody, a marker of the monocyte-macrophage lineage) were used to follow osteoclast differentiation quantitatively as a function of time after activation of resorption, from day 0 to day 4 (peak of resorption in this model). Local cell proliferation was assessed in parallel. Between day 0 and day 3, the thickness of the osteogenic compartment decreased by 50% (p < 0.0002). In the osteogenic compartment, proliferating cell numbers fell by 80% at 12 day, NSE(+) cells (located farthest from the bone surface) increased 3. 9-fold on day 4 vs. day 0 (p < 0.005), ED1(+) cells decreased between day 0 and day 2 (p < 0.02) before returning to their initial value, and TRAP(+) cells increased 2.7-fold between day 1 and day 3 (p < 0.0005). Resorption was absent in the site studied on day 0, but on day 4 there were 20.5 osteoclast nuclei per millimeter of bone surface. The cell ratio changed from 30.3 NSE(+) and ED1(+) (some of which were also TRAP(+)) cells per millimeter on day 0 to 37.6 mononucleated cells plus 20.5 osteoclast nuclei on day 4. In the nonosteogenic compartment, an entry of ED1(+)/NSE(-) was observed on 12 day (+23 cells, p < 0.02 vs. day 0). This was followed by a return of ED1(+) cell numbers to the control level on day 1, and a transient increase in NSE(+) cells (+47% on day 2 vs. day 1, p < 0.02). TRAP(+) cells were never seen in this compartment. Proliferating cell numbers did not change throughout the study. Our results strongly suggest that the osteoclasts present on day 4 differentiated from the pool of TRAP(+), ED1(+), and NSE(+) cells present at the site on day 0. The osteogenic compartment was gradually replenished by cells migrating from the nonosteogenic compartment, which was supplemented by ED1(+) cells recruited from the circulation early after activation. Moreover, osteogenic cells appeared to be as crucial in vivo for the acquisition of the TRAP phenotype as previously shown in vitro. PMID- 11062349 TI - Influence of nutritional factors on bone collagen fibrils in ovariectomized rats. AB - The influence of both vitamin D(3) and Ca:P ratio on bone collagen fibrils was investigated in ovariectomized rats. Six weeks after ovariectomy the rats were maintained for 80 days with diets containing vitamin D(3) and calcium supplementation. Age-matched ovariectomized animals were fed a normal diet. When vitamin D(3) was increased in the diet, although no effect in fibril organization was observed in relation to that from ovariectomized rats with the normal diet, a highly significant effect in fibril diameter was detected. When the calcium:phosphorus (Ca:P) ratio was increased from 1:1 to 2:1 (without vitamin D(3) supplementation) both structural fiber parameters were significantly affected. The results were closer to normal (i.e., collagen fibrils from animals without ovariectomy) when vitamin D(3) and Ca:P ratios were combined. PMID- 11062350 TI - Treatment with risedronate or alendronate prevents hind-limb immobilization induced loss of bone density and strength in adult female rats. AB - Immobilization leads to rapid loss of bone mass and mechanical competence, and long-term immobilization or repeated periods of short-term immobilization can have serious skeletal consequences and may lead to increased fracture liability. The aim of the present preclinical study was, therefore, to assess whether two antiresorptive agents, risedronate (Ris) or alendronate (Aln), would be capable of preventing immobilization-induced loss of bone mass and strength in rats. The study was designed as a dose-response study, and the site-specific effects of immobilization and of treatment are described. Four-month-old virgin female Sprague-Dawley rats were divided into eight groups with 12 animals in each group: (1) immobilized (Imm) control; (2) normal control; (3) Imm + Ris 0.1 mg/kg body weight/day (b.w./day); (4) Imm + Ris 0.2 mg/kg b.w./day; (5) Imm + Ris 1.0 mg/kg b.w./day; (6) Imm + Aln 0.2 mg/kg b.w./day; (7) Imm + Aln 1.0 mg/kg b.w./day; and (8) Imm + Aln 2.0 mg/kg b.w. /day. In groups 1 and 3-8, the right hind leg was immobilized with an elastic bandage. The study period was 28 days. The effects of unilateral hind-limb immobilization and of treatment were determined by dual energy X-ray absorptiometry (DEXA) measurements on tibiae and by biomechanical testing of femora at three different sites: diaphysis; femoral neck; and distal metaphysis. Bilateral measurements were performed (on the immobilized and nonimmobilized legs). Immobilization induced a significant loss of bone mineral density (BMD) at the proximal tibial metaphysis, but no change at the mid diaphysis. Furthermore, immobilization induced a loss of bone strength at the two femoral metaphyses, but no change was seen in three-point bending of the diaphysis. Both risedronate and alendronate treatment showed a dose-dependent protection against the immobilization-induced loss of bone density and strength at the metaphyses. We conclude that, in rats, short-term hind-limb immobilization affects only the metaphyses and that no changes are seen in the diaphysis. Both risedronate and alendronate can prevent immobilization-induced bone loss at the metaphyses. The present study confirms the importance of examining several skeletal sites when testing the efficacy of therapeutic agents. PMID- 11062351 TI - Additive effects of combined treatment with etidronate and alfacalcidol on bone mass and mechanical properties in ovariectomized rats. AB - The present study was undertaken to evaluate the effects of combined treatment with intermittent cyclical etidronate and daily alfacalcidol on the mass and the mechanical properties of bone in ovariectomized rats, and to compare the effects with those of single treatments. Seventy 14-week-old female rats underwent ovariectomy (ovx) or sham operation, and were assigned to seven groups (n = 10 each): sham-operated; ovx; ovx treated with etidronate; ovx treated with 0.1 microg/kg alfacalcidol; ovx treated with 0.2 microg/kg alfacalcidol; ovx treated with etidronate and 0.1 microg/kg alfacalcidol; and ovx treated with etidronate and 0.2 microg/kg alfacalcidol. One week after the operation, etidronate (4 mg/kg per day) was intermittently injected into rats for 2 weeks followed by a 10 week period of no treatment, and alfacalcidol was administered orally every day. After 24 weeks of treatment, all single and combined treatments increased the bone mineral densities (BMDs) of the proximal tibiae, midfemurs, and the fourth and fifth vertebral bodies, which had been decreased by ovx. Combined treatment groups showed higher BMDs than single treatment groups, and the effects were almost equal to the addition of those of respective single treatment groups. The combined treatment also showed additive effects on the mechanical properties of both midfemurs and L4 vertebral bodies. The increases in mechanical properties were proportional to those in BMDs. Analyses of microcomputed tomography images and histology confirmed the strong effects of combined treatments on both trabecular and cortical bone mass without impairment of mineralization or connectivity. We conclude that the combined treatment with etidronate and alfacalcidol additively increases the mass of bone with normal quality, resulting in bone strengthening in ovx rats. PMID- 11062352 TI - Development of bone canaliculi during bone repair. AB - We recently found that silver impregnation staining with protargol (silver protein), that is, a modified Bodian method, is useful for histologically identifying the details of bone canaliculi structure, using thin sections of decalcified bone tissues. With this staining method, we conducted the present study to assess the development of bone canaliculi during the process of intramembranous ossification using a fracture-like stimulation model of the rat femur. After making a drill-hole in the cortex of the rat femur, decalcified thin sections were obtained after 3, 5, 7, and 14 days by the standard paraffin embedding procedure. Silver staining for bone canaliculi was performed using our previously reported technique. The results showed that woven bone covered the fracture surface of the cortex after 5 days, then immature lamellar bone attached to the woven bone after 7 days, and finally the lamellar bone matured and became thick with appositional growth after 14 days. The osteocytes in the woven bone appeared at an early stage of bone repair and developed a few canaliculi that were short and irregularly distributed in the osteoid matrix, while the osteocytes in the lamellar bone at a late stage formed many bone canaliculi that were long and regularly distributed in mature bone matrix. Therefore, we concluded that woven bone osteocytes may be necessary for induction of the lamellar bone osteocytes followed by active appositional growth of the lamellar bone at the early stage of bone repair, and also that both bone tissues could be clearly distinguished from one another based on the pattern of development of bone canaliculi by the osteocytes, as seen with the use of our sensitive staining method. PMID- 11062353 TI - A murine model of distraction osteogenesis. AB - Distraction osteogenesis is both a valuable clinical technique and a useful tool for investigating the basic mechanisms involved in bone tissue regeneration. Here we describe the development of a murine model of this procedure that can be used in transgenic animals to investigate the role of specific genes in tissue regeneration. Ring fixators were applied to the lower leg of 12 normal adult male mice. An osteotomy was made in the diaphysis of the tibia, and 7 days after the operation the bone fragments were distracted by 0.25 mm twice a day for 10 days. Specimens were examined immediately at the end of distraction and after 14-70 days of consolidation. At the end of distraction, the distraction gap was filled with fibroblast-like cells arranged longitudinally. After 14 days of consolidation, there was radiographical evidence of bone formation in the distraction gap and, after 28 days of consolidation, the bone fragments were fused with regenerated bone. By 70 days of consolidation, the regenerated bone had been almost completely remodeled and the intramedullary canal reestablished. This study is the first to report consolidation of the distraction gap with regenerated bone in a murine model of distraction. PMID- 11062354 TI - Erect bipedal stance exercise partially prevents orchidectomy-induced bone loss in the lumbar vertebrae of rats. AB - This study investigates the responses of the fourth and fifth lumbar vertebral bodies of 6-month-old male Sprague-Dawley (SD) rats to orchidectomy (orx) and to erect bipedal stance for feeding for 12 weeks in specially designed raised cages (RC) for which the heights were raised from 20 cm to 35.5 cm. A total of 30 rats were divided into groups of: baseline; sham + housed in normal height cage (NC); orx + NC; sham + RC; and orx + RC. Bone histomorphometry was performed on the triple-labeled undecalcified fourth sagittal (LVL-4) and fifth transverse (LVX-5) sections. We found that orchidectomy induced high-turnover trabecular and cortical bone loss in the lumbar vertebrae. Forcing the rats to rise to erect stance for feeding reduced trabecular and cortical bone loss caused by orx. Apparently, depressing the elevated bone resorption next to the marrow induced by orx, and stimulating bone formation at the ventral periosteal surfaces, caused these effects. Orchidectomy and raised cage had similar effects on the two vertebrae except that the percentage of trabecular bone loss was greater in the LVL-4 than in LVX-5, and that bipedal stance exercise increased the total tissue area and mineral apposition rates (0-80 day interval) of ventral periosteal and dorsal endocortical surfaces of LVX-5 to a greater extent than it did in LVL-4. Such findings suggest that forcing rats to rise to an erect bipedal stance for feeding helps prevent loss of trabecular and cortical bone "mass," and presumably bone strength, in orchidectomized rats. This method also provides an inexpensive, noninvasive, reliable model to increase in vivo vertebral loading in rats that is similar in humans. PMID- 11062355 TI - Alendronate prevents bone loss in Chinese women with osteoporosis. AB - The objectives of the Hong Kong study are to investigate the efficacy of 10 mg alendronate in preventing bone loss at the hip and spine in osteoporotic Chinese women. One hundred osteoporotic Chinese women, aged 60-79 years, were randomized to receive 10 mg of alendronate or placebo, with 500 mg elemental calcium. Bone mineral density (BMD) at the spine and hip were measured at baseline, 6 months, and 12 months. Seventy-eight subjects completed the study. The alendronate treated group gained more bone at both the spine (p < 0.01) and femoral neck (p < 0.001), with a mean difference (+/-SE) of 2.4% (+/-0.86%) at the spine and 3.98% (+/-0.95%) at the femoral neck. Of the 100 patients, 6 subjects in the alendronate group and 5 subjects in the placebo group had mild gastrointestinal symptoms. We conclude that alendronate (10 mg) was effective in preventing bone loss in postmenopausal osteoporotic Chinese women. PMID- 11062356 TI - Effects of alendronate on osteopenic postmenopausal Chinese women. AB - To evaluate the effects of alendronate on postmenopausal Chinese women with osteopenia, we treated 46 subjects daily with either 10 mg alendronate (N = 24) or placebo plus 500 mg calcium supplement (N = 22), and measured their bone mineral density (BMD) at the lumbar spine and hip, and urinary bone resorption markers before, during, and after the 1 year treatment period. The bone markers included N-telopeptide of type I collagen (NTx) and deoxypyridinoline (Dpd); both were corrected by the concentration of creatinine in the same sample (NTx/Cr and Dpd/Cr). Both NTx/Cr and Dpd/Cr decreased significantly by 44% and 28%, respectively (p < 0.05 for both), in 1 month in the active treatment group but did not change in the placebo group. BMD at the spine, femoral neck, trochanter, and Ward's triangle increased significantly by 6 months and showed a further increase through month 12 at the spine in the alendronate-treated group. Relative to the placebo group, BMD changes at various sites in the alendronate-treated group were higher at 12 months by 6%-11%. Thus, our data suggest that 10 mg alendronate daily resulted in significant increases in spine and hip BMD, and decreases of urinary resorption markers in the osteopenic postmenopausal Chinese women studied. The amplitude of responses was higher than in previous reports in the USA and Europe. PMID- 11062357 TI - Alendronate increases bone strength by increasing the mean degree of mineralization of bone tissue in osteoporotic women. AB - The mean degree of mineralization of bone (MDMB) was measured by quantitative microradiography on transiliac bone biopsies taken from 53 postmenopausal osteoporotic women who had been treated with alendronate (ALN; 10 mg/day) during 2 (9 patients) or 3 years (16 patients) or with placebo (PLA; 15 and 13 patients, respectively). In the same patients, bone mineral density (BMD) values were obtained by dual-energy X-ray absorptiometry of the lumbar spine and femoral neck at the beginning and end of treatment. Histomorphometric parameters and activation frequency of new remodeling units were also measured on the iliac biopsies. After 2 years of ALN, MDMB in compact bone was 9.3% (p = 0.0035) and in cancellous bone was 7.3% (p = 0.0009) higher, respectively, than PLA. After 3 years of ALN, MDMB in compact bone was 11.6% (p = 0.0002) and in cancellous bone was 11.4% (p = 0.0001) higher, respectively, than PLA. After 2 and 3 years of ALN, and compared with the corresponding PLA, the distribution of the degree of mineralization in compact and cancellous bone showed a clear shift toward the highest mineralization values and a decrease in the number of bone structure units having low values of mineralization. The between-group differences in MDMB were similar to those of BMD at the lumbar spine BMD (+8.7% after 2 years and +9.6% after 3 years, respectively), suggesting that MDMB augmentation probably accounted for the majority of the increase in BMD seen with ALN. The data support the hypothesis that the reduction in activation frequency caused by the antiresorptive effect of ALN is followed by a prolonged secondary mineralization that increases the percentage of bone structure units having reached a maximum degree of secondary mineralization and, through this mechanism, MDMB. That these effects contribute to improved bone strength is demonstrated by the reduction in fracture incidence previously demonstrated in these patients. PMID- 11062358 TI - Fractures following thyroidectomy in women: a population-based cohort study. AB - Hip fracture risk has been associated with hyperthyroidism and thyroidectomy in men and with hyperthyroidism in women, but the influence of thyroidectomy on fracture risk in women has not been adequately addressed. The 630 Rochester, MN women who underwent thyroidectomy in 1950-1974 were followed subsequently for 12,804 person-years (retrospective cohort study) during which 601 fractures were observed. Relative to incidence rates in the community, there was no increase in overall fracture risk (standardized incidence ratio [SIR] 0.9; 95% confidence interval [CI] 0.8-1.00). No increase was seen in limb fractures generally or in distal forearm fractures specifically (SIR 1.1, 95% CI 0.8-1.4). There was a modest but statistically significant increase in the risk of hip fractures following thyroidectomy (SIR 1.3, 95% CI 1.01-1.8), but much greater increases were apparent in the risk of subsequent fractures of the ribs, spine, and pelvis. There was almost a threefold increase in vertebral fractures (SIR 2.8, 95% CI 2.3 3.3), but the excess was mostly observed long after the original operation and may be attributable to ascertainment bias. Fracture risk was associated with advancing age and with the presence of one or more of the diseases that have been associated with secondary osteoporosis but not with a history of hyperthyroidism, extent of thyroid surgery, or subsequent use of thyroid replacement therapy. Thus, with the exception of some fractures of the axial skeleton, which might have been more completely diagnosed among affected women, there was no increase in fracture risk among women following thyroidectomy performed mainly for adenoma or goiter. PMID- 11062359 TI - Ambulatory level and asymmetrical weight bearing after stroke affects bone loss in the upper and lower part of the femoral neck differently: bone adaptation after decreased mechanical loading. AB - The aim of this 1-year prospective study of acute stroke patients was to determine the effects of walking and asymmetrical weight bearing on the loss of bone mineral in the upper and lower femoral neck. Forty patients were followed. Eight remained unable to walk, whereas 32 relearned to walk independently within 7 months (12 shortly after the stroke, 15 by 2 months, 5 by 7 months). Bone mineral density (BMD) was measured in the proximal femur within the first week after stroke and 1 year later; regional BMD changes were computed for the lower and upper femoral neck. The lower part of the femoral neck is mainly influenced by compressive stresses of the hip, the upper part by tensile stresses during walking. When comparing mean BMD loss in groups of patients according to when they relearned to walk, a statistically significant trend in BMD loss was found in the lower femoral neck on both the paretic and nonparetic sides (p < 0.01 and p = 0.01, respectively), whereas, for the upper femoral neck, no significant trend was seen (p >/= 0.1). In addition, the body weight distribution during standing was assessed by use of a force-plate in 38 patients who could stand independently at the 7 month evaluation. The only significant correlation between changes in BMD and asymmetrical weight bearing was found in the lower femoral neck on the paretic side (r = 0.6, p < 0.001). In conclusion, this study shows that the reduction in BMD in the femoral neck occurs mainly in the lower part of the neck and on the paretic side. The BMD loss depended on when or if the patients relearned to walk, but also on the amount of body weight born on the paretic leg. Thus, measuring the lower part of the femoral neck gives a better estimate of the impact of gait and weight bearing than measuring the total femoral neck. PMID- 11062360 TI - Contributions of bone density and geometry to the strength of the human second metatarsal. AB - We investigated, at the whole bone level, the contribution of bone density and geometry to the fracture load of the second metatarsal, a bone that is prone to stress fracture. Dual-energy X-ray absorptiometry (DXA) was used to determine the areal bone mineral density (BMD), projected area of bone, and bone mineral content. Peripheral quantitative computed tomography (pQCT) was used to determine the volumetric cortical bone mineral density (vCtBMD) and cross-sectional moment of interia. Various metatarsal linear dimensions were also measured. The load at failure in cantilever bending was determined. The only linear dimension that had a significant correlation with load at failure was the height of the metatarsal base (r(2) = 0.30, p = 0.008). Utilizing all of the information provided by DXA gave no greater indication of whole bone strength than just BMD alone (adjusted r(2) = 0.40, p = 0.001). Using all of the information provided by pQCT gave no greater indication of whole bone strength than just vCtBMD alone (r(2) = 0. 46, p < 0.001). Volumetric cortical density and BMD were strongly correlated (r(2) = 0.81, p < 0.001). Our data suggest that, in the human second metatarsal, a variable such as material strength (as inferred from cortical density), and not geometry, may be the major factor in determining cantilever load to failure. PMID- 11062361 TI - Parallel plate model for trabecular bone exhibits volume fraction-dependent bias. AB - Unbiased stereological methods were used in conjunction with microcomputed tomographic (micro-CT) scans of human and animal bone to investigate errors created when the parallel plate model was used to calculate morphometric parameters. Bone samples were obtained from the human proximal tibia, canine distal femur, rat tail, and pig spine and scanned in a micro-CT scanner. Trabecular thickness, trabecular spacing, and trabecular number were calculated using the parallel plate model. Direct thickness, and spacing and connectivity density were calculated using unbiased three-dimensional methods. Both thickness and spacing calculated using the plate model were well correlated to the direct three-dimensional measures (r(2) = 0. 77-0.92). The correlation between trabecular number and connectivity density varied greatly (r(2) = 0.41-0.94). Whereas trabecular thickness was consistently underestimated using the plate model, trabecular spacing was underestimated at low volume fractions and overestimated at high volume fractions. Use of the plate model resulted in a volume-dependent bias in measures of thickness and spacing (p < 0.001). This was a result of the fact that samples of low volume fraction were much more "rod like" than those of the higher volume fraction. Our findings indicate that the plate model provides biased results, especially when populations with different volume fractions are compared. Therefore, we recommend direct thickness measures when three-dimensional data sets are available. PMID- 11062362 TI - Update on transrectal ultrasound-guided needle biopsy of the prostate. AB - Over the past decade, the sextant biopsy technique has emerged as the standard of care in the detection of prostate cancer. This technique is easy to learn and well tolerated by patients and has a major complication rate of <1%. However, limitations in cancer detection have been appreciated, particularly a false negative rate approaching 25%. This high failure rate has led investigators to refine biopsy techniques to improve cancer detection. Intuitively, increasing the total number of cores should improve cancer detection. However, the optimal core number has yet to be defined. Confounding factors include variability of prostate size, tumor volume, and tumor location. Currently, a new standard is emerging prescribing a minimum of eight cores, of which at least three are directed at the lateral aspect of the peripheral zone. These additional biopsies appear to enhance cancer detection by about 15%. The improved yield is most pronounced among patients with a serum prostate specific antigen concentration between 4 and 10 ng/mL and larger gland volume (>50 cc). These additional biopsies may decrease the need for repeat biopsies. In the meantime, strategies are being developed for the optimal technique of repeat biopsies among patients with persistent clinical suspicion in the setting of a prior negative biopsy. Currently, recommendations include increasing the biopsy number to a minimum of 10 cores, including sampling of the lateral peripheral and transition zones. PMID- 11062363 TI - Immunohistochemical changes in prostate cancer after androgen deprivation therapy. AB - Androgen deprivation induces substantial changes in the phenotype of prostate cancer that are accompanied by alterations in protein expression. Immunohistochemical studies allow precise cellular localization of such expression, thereby providing an understanding of the biochemical alterations caused by therapy. Expression of proteins may be increased (e.g., multiple growth factors, heat shock protein), decreased (e.g., microvessel density, proliferation markers, certain integrins), or remain unchanged (e.g., prostate specific antigen, prostatic acid phosphatase, prostate-specific membrane antigen, and other secretory proteins). Variations in immunoreactivity may be of prognostic value in some patients. This report summarizes the existing literature regarding changes in tissue expression of proteins, as determined by immunohistochemistry, and the clinical implications of these changes. PMID- 11062364 TI - Epidemiology and molecular biology of early prostatic neoplasia. AB - This paper provides our institutional data with respect to the prevalence of early neoplastic changes within the prostate gland and the age and race distribution of the patients. The changes examined were prostatic intraepithelial neoplasia (PIN) and preclinical (latent) cancers. The literature on the prevalence of these early lesions among different geographic and ethnic groups is summarized, and an abbreviated review of the more common molecular alterations reported at this early phase of prostatic neoplasia is offered. PMID- 11062365 TI - Difficulties in interpreting specimens after neoadjuvant hormonal therapy and radiation with illustration of neuroendocrine differentiation. AB - Pattern and cellular changes attributable to neoadjuvant hormonal therapy (NHT) might cause the unwary pathologist to overgrade or fail to recognize a treated prostatic cancer. Overdiagnosis and overgrading of surgical resections and biopsies can be avoided if an appropriate history of therapy is conveyed with the surgical specimen and if the pathologist is aware of the altered morphology of prostatic cancer treated by NHT alone or NHT plus radiation. Study of three prostatectomy specimens with post-NHT predominance of neuroendocrine cells showed positive staining for prostate specific antigen (PSA) and prostatic acid phosphatase (PAP), as well as staining for chromogranin and synaptophysin in Paneth-like and small neuroendocrine cells. Difficult-to-interpret needle biopsies and transurethral resection (TUR) biopsies of prostate, where the urologic pathologist's suspicion of a radiation effect was confirmed by additional history, showed absence of the basal cell layer with 34 beta E12 keratin immunostaining in prostatic cancer glands, while basal cells were present in the nonneoplastic glands with radiation-induced atypia. Postradiation salvage prostatectomy specimens showed greater apoptosis after combined NHT and radiation than after radiation without NHT. Changes attributable to radiation and radiation plus NHT are illustrated. PMID- 11062366 TI - Significance of the Gleason scoring system after neoadjuvant hormonal therapy. AB - Neoadjuvant hormonal therapy (NHT) induces morphologic changes in prostate adenocarcinoma that result in the assignment of higher Gleason scores on average than in pretreatment biopsy specimens. This outcome has led to the recommendation that the Gleason scoring system not be applied to prostate adenocarcinoma specimens after NHT. We reviewed the radical prostatectomy specimens of 116 patients who had received NHT. Gleason scores were assigned on the post-treatment specimens by applying the usual criteria; in addition, an estimated pretreatment Gleason score was assigned on the basis of knowledge of the morphologic alterations associated with NHT. Finally, an estimate of the degree of therapy effect was assigned: little or no evidence of hormonal effect (grade 1) to marked therapy-related changes (grade 3). Both the post-treatment and the estimated pretreatment Gleason score correlated significantly with biochemical progression (P = 0.03 and P = 0.03, respectively; log-rank test). The degree of therapy effect did not correlate with progression (P = 0.46; log-rank test). This limited analysis suggests that despite the morphologic alterations induced by NHT, post treatment Gleason score remains a significant prognostic measure. Further studies in more uniformly treated populations are required to confirm this observation. PMID- 11062367 TI - Three-dimensional ultrasound analyses of the prostate. AB - Although conventional ultrasonography has proven to be clinically useful for depicting many types of cancerous lesions, it cannot distinguish reliably between cancerous and noncancerous tissue of the prostate. Therefore, conventional transrectal ultrasonography (TRUS) is used primarily for general evaluations of the gland and for guiding biopsies based on clearly imaged anatomic features such as the capsule, seminal vesicles, and urethra. Spectrum analysis extracts ultrasound signal parameters associated with biopsy-proven tissue types, and these parameters are then classified using neural network tools such as learning vector quantization, radial basis, and multilayer perceptron algorithms. Classification of cancerous and noncancerous prostate tissue using neural networks produces receiver operating characteristic (ROC) curves of 0.87 +/- 0.04 compared with 0.64 +/- 0.04 for conventional ultrasonography. To image the prostate using these methods, parameter values are computed at each pixel location, then translated into a score for the likelihood of cancer using a look up table generated using the best classification algorithm. The score for cancer likelihood is expressed as a gray-scale or color value, and the resulting image may be useful to guide biopsies or therapy. Changes in parameter or score values over time potentially can be used to assess progression of disease or efficacy of therapy. PMID- 11062368 TI - Magnetic resonance spectroscopic studies of the prostate. AB - Since the first suggested use of nuclear magnetic resonance (NMR) for detecting cancer, followed by the demonstration of the feasibility of imaging based on the NMR signal in 1973, magnetic resonance imaging (MRI) has become the modality of choice for a variety of clinical applications. Subsequently, the use of NMR spectroscopy (MRS) to detect the presence of different metabolites in vivo has provided unique opportunities for obtaining physiological and biochemical information. More recently, improvements in NMR equipment (magnet, electronics, computers, gradients coils, radiofrequency coils) and pulse sequences (software) have further improved these capabilities. The distinctions between MRI and MRS have begun to blur as new techniques emerge that combine imaging and spectroscopy, generating MRS images of a variety of metabolites. This review provides a brief overview of recent developments in MRS studies pertinent to the clinical evaluation of prostate cancer. The paper has been divided into three parts: a brief qualitative theoretical section about MRS, a review of in vitro studies, and a discussion of the clinical studies of the human prostate. PMID- 11062369 TI - Short-course androgen ablation combined with external-beam radiation therapy and low-dose-rate permanent brachytherapy in early-stage prostate cancer: a matched subset analysis. AB - BACKGROUND AND PURPOSE: In order to evaluate the effect of short-term androgen blockade on biochemical control rates for high-risk patients receiving a combination regimen of external-beam radiation therapy and low-dose-rate permanent seed implant brachytherapy, a retrospective matched subset analysis was performed. PATIENTS AND METHODS: Inclusion in the high-risk cohort required at least two of the following poor prognostic factors: serum prostate specific antigen (PSA) concentration > or = 10.0 ng/mL, Gleason score > or = 7, or clinical stage T(2c) or T(3a) disease. Twenty-one patients who underwent androgen ablation between June 1991 and December 1995 in addition to combined-modality radiation therapy qualified as high risk, as did 77 patients who underwent combined-radiation therapy only. There was no statistically significant difference between the two groups in terms of follow-up (mean 44.6 v 47.8 months, respectively), pretreatment PSA, clinical stage, biopsy Gleason score, or the presence of all three poor prognostic factors. RESULTS: The overall rates of freedom from biochemical failure at 5 years were 77% in the hormonally treated group and 58% in the nonhormonally treated group. The difference was not statistically significant by log rank test (P = 0.08). CONCLUSION: Longer follow up with larger patient numbers is needed to define the role of adjuvant androgen ablation combined with radiation therapy. PMID- 11062370 TI - Effects of neoadjuvant hormonal therapy on prostate biopsy results after (125)I and (103)Pd seed implantation. AB - BACKGROUND: Androgen ablation may improve the efficacy of radiation therapy. PATIENTS AND METHODS: A total of 296 patients who had either (125)I (206; 70%) or (103)Pd (90; 30%) transperineal prostate brachytherapy (no external-beam radiation) had routine transrectal ultrasound-guided needle biopsy (minimum six cores) 2 years after treatment without regard to disease status. Neoadjuvant hormonal therapy (NHT: leuprolide acetate and flutamide) was used in 115 patients (39%) for 3 months prior to and 3 months after the implant. RESULTS: Of the 296 patients, 30 (10%) had positive prostate biopsies. Biopsies were positive in 4 of 115 (3.5%) v 26 of 181 (14%) of those who received or had not received NHT, respectively (P = 0.002). When patients were separated into low risk (PSA < or = 10 ng/mL, stage < or = T(2a), and Gleason score < or = 6) and high risk (all others), it was seen that low-risk patients did not benefit from NHT (3.8 v 7.7% positive biopsy rate; P = 0.5) whereas high-risk patients did (3.4% v 21.1%; P = 0.003). CONCLUSION: Prostate brachytherapy yields high negative biopsy rates (90%) 2 years after treatment. Neoadjuvant hormonal therapy can improve the local control rates (as determined by biopsy) in patients undergoing (125)I or (103)Pd seed implantation. These results are most significant for patients who present with PSA >10 ng/mL, stage > or = T(2b) diseases, or Gleason score > or = 7 (high risk status). PMID- 11062371 TI - Clinical utility of percent-positive prostate biopsies in predicting biochemical outcome after radical prostatectomy or external-beam radiation therapy for patients with clinically localized prostate cancer. AB - BACKGROUND AND PURPOSE: The clinical utility of the percentage of positive prostate biopsies in predicting prostate specific antigen (PSA) outcome after radical prostatectomy (RP) or external-beam radiation therapy (EBRT) for men with PSA-detected or palpable prostate cancer is not established. METHODS: A Cox regression multivariable analysis was used to determine whether percent-positive prostate biopsies provided clinically relevant information about PSA outcome after RP in 960 men, while accounting for the previously established risk groups based on the pretreatment PSA concentration biopsy Gleason score, and the 1992 American Joint Commission on Cancer clinical T stage. RESULTS: In the intermediate-risk group, 80% of the patients (stage T(2b) or biopsy Gleason 7 or PSA 10-20 ng/mL) could be classified into either an 11% or an 86% 4-year PSA control cohort using the preoperative prostate biopsy data. These findings were validated using an independent surgical (N = 823) and radiation (N = 473) data set. Percent-positive prostate biopsies added clinically significant information regarding time to PSA failure after RP. CONCLUSIONS: The percentage of positive prostate biopsies should be considered in conjunction with the PSA level, biopsy Gleason score, and clinical T stage when counseling patients with newly diagnosed and clinically localized prostate cancer about PSA outcome after RP or EBRT. PMID- 11062372 TI - High-intensity focused ultrasound in prostate cancer: results after 3 years. AB - BACKGROUND: Local high-intensity focused ultrasound (HIFU) is a minimally invasive method of coagulation (85 degrees C) that ablates prostatic tissue with high precision. PATIENTS AND METHODS: Over a 3-year period, 184 patients with organ-confined prostate cancer have undergone 232 sessions of transrectal HIFU therapy (mean duration 90 minutes) under spinal anesthesia at 2.25 or 3.0 MHz, 50 W, with a penetration depth of 25 mm. RESULTS: Follow-up sextant biopsies (mean 1.9) were cancer free in 80% of patients, and in patients with residual cancer, the tumor mass was reduced more than 90%. The nadir value of prostate specific antigen (PSA) was <4 ng/mL in 97%, including 61% who had values <0.5 ng/mL. After primary HIFU, no severe side effects (fistula, grade 2 or 3 incontinence, rectal mucosal burn) were seen. All patients had a suprapubic tube (mean 29 days), and 33% needed transurethral resection of debris (mean 7 g). Hospital discharge was within 23 hours after treatment. CONCLUSION: Transrectal HIFU enables minimally invasive local prostate tissue ablation with high rates of negative biopsies, low PSA nadir, and low complication rate. Further follow-up is needed to define the efficacy of disease control. PMID- 11062373 TI - High-intensity focused ultrasound: complications and adverse events. AB - BACKGROUND: Transrectal high-intensity focused ultrasound (HIFU) is under investigation as a minimally invasive therapeutic option for elderly men with prostatic cancer. METHODS: From November 1996 to April 1999, 315 HIFU treatments with the Ablatherm (EDAP/TMS) were performed. A questionnaire including 50 theoretically possible adverse events was developed. Every patient complaint was recorded, including the physician's and patient's assessment, before and after therapy. Start date, end date, and period of every complaint were analyzed. RESULTS: As major adverse events after primary HIFU, there were six cases of stress incontinence grade 1. After repeat treatments with HIFU, rectourethral fistulas occurred in five patients, stress incontinence grade 1 in eight, and, after additive transurethral resection, grade 2 in one and grade 3 in two patients. Post-HIFU rectal mucosa burn decreased from 15% in 1996 to 0 within the last year. In all treatments, obstruction was avoided by suprapubic urinary diversion. Urinary tract infections (UTIs) were recorded initially in 58% of patients but later in only 8%. CONCLUSION: Transrectal HIFU proved to be a secure, minimally invasive therapeutic option for elderly men to avoid hormonal ablation or to postpone its first use. PMID- 11062374 TI - Intermittent androgen suppression for prostate cancer: Canadian Prospective Trial and related observations. AB - The Canadian Prospective Trial of intermittent androgen suppression was a prototype therapeutic initiative started in 1995 for the management of patients in biochemical relapse after radiation for localized prostate cancer. An interim analysis has yielded several observations on the relations between baseline serum prostate specific antigen (PSA), nadir serum PSA, Gleason score, and time off treatment. In a typical androgen-dependent tumor, the response of serum PSA to androgen withdrawal is biphasic, but with early tumor progression, plateauing of serum PSA is observed. Ligand-independent activation of the androgen receptor, a mechanism subserving the initiation of androgen independence, can be counteracted experimentally with decoy molecules and clinically with nonsteroidal antiandrogens. In some patients, it is possible to lengthen the off-treatment interval by inhibiting the enzyme 5 alpha-reductase, an effect that can be reinforced by lowering serum testosterone with an antigonadotropin. Serial measurements of serum PSA indicate that intermittent androgen suppression engenders a more diverse range of hormone-related responses than previously appreciated. These include: (1) repeated differentiation of tumor with recovery of apoptotic potential; (2) inhibition of tumor growth by rapid restoration of serum testosterone; and (3) restraint of tumor growth by subnormal levels of serum testosterone. These responses are aspects of regulation that should be taken into account when planning long-term treatment of prostate cancer with intermittent androgen suppression. PMID- 11062375 TI - The case for neoadjuvant androgen suppression before radiation therapy. AB - Neoadjuvant androgen suppression (NAS) can reduce the number of tumor clonogens prior to radiation, thus increasing the tumor control probability. Also, NAS may sensitize tumor cells to radiation if cell kill by both modalities follows a common pathway. The timing and sequence of NAS and radiation are important, with radiation being most effective if given at the point of maximal tumor regression. The biologic rationale for NAS + radiation has been reinforced by results from randomized trials, in particular RTOG 8610. However, many murine adenocarcinomas respond to androgen deprivation by a reduction in the proliferation rate and arrest in G(0), and in vitro data suggest that this arrest may interfere with radiation-induced cell killing. The mechanism of cell killing after low-dose-rate radiation (brachytherapy) may be different from that after high-dose-rate treatment. There are no reported experimental data assessing the interaction of NAS and brachytherapy to determine whether the combination offers a theoretical advantage or is potentially deleterious. Whether we understand the mechanism or not, clinical trials seem to support a positive interaction of NAS with external beam radiation, but we have only begun to explore the timing and sequence that will provide the maximal effect. It cannot be assumed that the same advantage will hold with brachytherapy. PMID- 11062376 TI - Combined androgen deprivation with radiotherapy for prostate cancer: does it make sense? AB - Currently, our understanding of the mechanism(s) of radiation-induced death of prostate cancer is limited. In-depth analysis of these processes would facilitate the design of more effective treatment strategies utilizing radiation therapy. An increasingly recognized form of radiation-induced death is postmitotic apoptosis. In this process, radiation damages the tumor cell s DNA. The cell then divides prior to completing DNA repair, an event that is lethal. In order to avoid this fate, the cancer cell may attempt to halt its cell-cycle machinery temporarily to repair its DNA prior to dividing. In the treatment of prostate cancer, radiation therapy currently is being evaluated in combination with androgen deprivation (AD). However, because AD can induce growth arrest, it may reduce the effectiveness of radiation through a reduction in postmitotic apoptosis. To study this effect, we examined the effect of AD on prostate cancer radiosensitivity as it is related to cell-cycle progression. Androgen-sensitive prostate cancer cells demonstrated increased resistance to radiation when deprived of androgenic stimuli. Thus, paradoxically, AD may reduce the radiosensitivity of prostate cancer by means of cell-cycle delay, which results in a reduction in postmitotic apoptosis. PMID- 11062377 TI - Overview of evolving strategies incorporating prostate-specific membrane antigen as target for therapy. AB - Prostate-specific membrane antigen (PSMA) is a potential target in prostate cancer patients because it is very highly expressed and because it has been reported to be upregulated by androgen deprivation. This overview addresses the expression of the PSMA gene in terms of the promoter and enhancer and how that may play a role in gene therapy. We also review PSMA as a target for antibodies for imaging and treatment and the development of a novel hybrid T-cell receptor that combines the specificity of anti-PSMA antibodies with that of T-cell receptor activation when introduced into primary lymphocytes by retroviral mediated gene transfer. We also discuss our recent findings on the expression of a PSMA-like gene and how that understanding allows specific targeting of PSMA. PMID- 11062378 TI - Resistance to apoptosis in prostate cancer cells. AB - Androgen-independent prostate cancer cells are remarkably resistant to therapeutic agents that work by triggering apoptosis via the caspase cascade. The recent sequencing of the entire genome of one of the most radiation-resistant organisms known, Deinococcus radiodurans, yields some insight into how prostate cancer cells might mount such resistance to apoptosis. Rather than being attributable to any one mechanism, the extreme radiation resistance of D. radiodurans appears to reflect the expression of a large number of different systems capable of preventing, repairing, or tolerating DNA damage and a very high degree of redundancy in these systems. Many molecular alterations that may influence the threshold for apoptosis have already been described in advanced prostate cancer; changes in bcl-2, p53, and the androgen receptor have been the most extensively studied. Current information is consistent with the concept that individual prostate cancer cells express multiple antiapoptotic mechanisms. This conclusion implies that it will not be possible to enhance cellular sensitivity to therapeutics that activate apoptosis by disabling just one target in a pathway, because other proteins are likely to be available to assume its function. Likewise, even elimination of a whole pathway may have little effect on sensitivity because cellular viability is protected by so many different mechanisms. However, where molecular changes have a phenotypic consequence, they offer a window of opportunity for the development of novel therapeutic strategies. One such example is a recently identified small organic compound that can inhibit p53 function and thus protect normal tissues against radiation induced apoptosis without impairing killing of p53-deficient tumor cells. PMID- 11062379 TI - Neoadjuvant hormone therapy: the Canadian trials. AB - The Canadian Urologic Oncology Group has carried out three studies of neoadjuvant hormonal therapy (NHT) in prostate cancer. The first, a study of 3 months of cyproterone acetate (CPA) 100 mg TID in patients undergoing external-beam radiation therapy, showed a benefit with respect to time to biochemical progression. There are no survival or clinical progression data available from this study. The second study involved 3 months of CPA prior to radical prostatectomy compared with radical prostatectomy alone and enrolled 200 patients. The probability of biochemical progression at 36 months was similar in the two groups (CPA 40%; surgery alone 30%; P = 0.3233). More recently, we have carried out a randomized trial of 3 v 8 months of leuprolide plus flutamide prior to radical prostatectomy in 547 patients. Patients were stratified by clinical stage, Gleason grade, and serum prostate specific antigen (PSA) concentration. In the 3- and 8-month groups, presurgery PSA concentrations were <0.1 ng/mL in 35% v 73%, and >0.3 ng/mL in 37% v 10%, respectively. In the 3- and 8-month groups, the positive margin rates were 17% and 5% and the organ-confined rates 71% and 91% (P < 0.01). One-year follow-up is now available on the entire cohort. Data regarding time to biochemical and clinical progression and overall and disease-specific survival will be required to determine whether this change in the pathologic findings translates into a patient benefit. PMID- 11062380 TI - Update on Memorial Sloan-Kettering Cancer Center studies of neoadjuvant hormonal therapy for prostate cancer. AB - PURPOSE: We report the results of surgery in 520 patients with clinically localized carcinoma of the prostate (CaP) who received preoperative neoadjuvant hormonal therapy (NHT) for 3 to 11(+) months. METHODS: The results in the NHT patients were compared with those in 1,413 men having surgery without NHT at our institution during the same time period. In the group without pretreatment, the median and mean follow-up was 36 and 21 months, respectively. In the patients receiving NHT, the median follow-up was 33 months and the mean 41 months. RESULTS: The overall disease-free survival (DFS) rate (serum prostate specific antigen [PSA] concentration < or = 0.2 ng/mL) was 75% at 5 years and 50% at 10 years. There was no statistically significant difference in overall DFS rate between men who had NHT and those who did not. No DFS advantage could be demonstrated for those patients with a presenting PSA >20 ng/mL who received NHT compared with patients with the same PSA concentration who did not receive NHT. Despite our previous experience indicating improved survival with NHT in men with a presenting PSA of > 10 ng/mL, we could find no advantage to NHT in enhancing DFS. At a median survival of 35 months (mean 41 months) in 201 men with an initial PSA > or = 10 ng/mL, 70% had an undetectable PSA concentration at 5 years compared with 72% at the same time point in men presenting with PSA <10 ng/mL. In the group expected to have the best surgical result; i.e. those men whose preoperative PSA was < or = 7 ng/mL, there was no DFS difference in men given NHT compared with those having no hormonal manipulation. Patients presenting with stage T(1) disease had a significantly better DFS than those with either T(2) or T(3) CaP. However, within each stage, the addition of NHT to surgery did not result in a higher DFS rate. The 5- and 10-year DFS rates for stage T(1) were 80% and 64%, for T2 disease 78% and 50%, and for T3 disease 67% and 50%. There was a statistically significant difference (P < or = 0.003) in survival between stage T(1) and stage T(2) disease, but no significant difference in DFS was noted in patients presenting with stage T(2) compared with T3 cancer (P = 0.431). Gleason score was not a significant predictor of durable DFS, and the addition of NHT did not improve the DFS within groups of patients with similar Gleason scores. Men with only one or two positive biopsy cores did significantly better than those with more than three positive cores (P = 0.06). There was a significant difference in DFS between men who had organ-confined disease and those with disease outside the gland (P = 0.0003). However, NHT did not improve DFS. The presence of positive surgical margins was a negative prognostic factor (P = 0. 001). Men who received NHT had a statistically lower positive margin rate (P = 0.001), but NHT did not increase the likelihood of a durable DFS (P = 0.175). The duration of NHT did not affect the DFS (P = 0.100 for <3 v >3 months). CONCLUSION: There appears to be no subset of men undergoing radical prostatectomy in whom the routine administration of NHT is beneficial despite the statistically significant improvement in the pathologic findings. PMID- 11062381 TI - Neoadjuvant hormonal therapy prior to radical prostatectomy: the European experience. AB - BACKGROUND AND PURPOSE: Neoadjuvant hormonal therapy (NHT) has been used for more than a decade for prostate cancer, but the results of clinical trials are only now becoming available, and the value of the treatment is not yet clear. The authors reviewed the results of the European randomized trials to increase our understanding of the role of this treatment. PATIENTS AND METHODS: We report the results of 402 patients with prostate cancer (220 clinical stage T(2) and 182 clinical T(3) tumor), of whom 192 were randomly assigned to NHT using an LHRH analog (goserelin) plus flutamide for a period of 3 months (NHT) and 210 underwent radical prostatectomy only (RP). RESULTS: "Pathologic downstaging" occurred in 15% and 7% of the NHT and the RP group, respectively (P < 0.01). Fifty of the 189 patients in the NHT group (26%) and 68 of the 209 patients in the RP group (33%) developed disease progression, as determined by rising serum prostate specific antigen (PSA) concentration. Regarding local disease progression, the advantage for the use of NHT approached but did not reach statistical significance:18 of 189 patients (10%) in the NHT group and 33 of 209 patients (16%) in the RP group (P = 0. 07). CONCLUSIONS: Although there was a trend in favor of the NHT group with respect to the number of patients with PSA progression and the number with local disease progression, it did not reach statistical significance. These results may be attributable to a true lack of benefit of adjuvant hormonal ablation or to a lack of statistical power to demonstrate a difference in a subset of patients who might benefit from this therapy. PMID- 11062382 TI - Comparative study of the clinical efficacy of two dosing regimens of flutamide. AB - PURPOSE: We performed a randomized trial to compare the efficacy and toxicity of a new dose of flutamide (500 mg QD) with the currently recommended dose (250 mg q8h) in the treatment of advanced prostate cancer. The primary endpoints were percent of patients having normalization of prostate specific antigen (PSA), time to normalization, and percent change from baseline. Secondary endpoints were quality of life and toxicity. PATIENTS: Altogether, 440 men aged 46 to 94 years (mean 71 years) with confirmed stage M(1) disease, documented PSA rise >0.2 ng/mL, ECOG status 0 to 2, no second neoplasm, no liver function tests > or = 1.5 fold normal values, and no previous treatment for metastatic disease were entered in the trial. RESULTS: The PSA normalized by week 12 in 71% of the patients receiving 500-mg dose and 75% of those receiving the standard dose. The percent change in PSA was 89% and 96%, respectively. The treatment groups were not significantly different with respect to the incidence of adverse events: 71% v 68% in the 500-mg and 250-mg arms, respectively (P = 0.337). CONCLUSIONS: When combined with castration, 500 mg of flutamide appears to be equally effective in lowering serum PSA and is not significantly more toxic than conventional dosing. The use of 500 mg QD instead of the standard 250 mg q8h would result in a cost savings of 30%. PMID- 11062383 TI - Hormonal therapy options for biochemical recurrence of prostate cancer after local therapy. AB - Recurrence after local prostate cancer treatment detectable only by a rise in serum prostate specific antigen (PSA) is a very common problem facing clinicians. Given that the majority of these men are relatively young and otherwise healthy, treatment of PSA-only recurrence requires approaches that not only improve survival but also preserve quality of life. For radical prostatectomy patients, a PSA-only recurrence is broadly defined as persistent or rising PSA in the postoperative period. For radiation-treated patients, the 1997 American Society for Therapeutic Radiology and Oncology guidelines specify three consecutive elevations of PSA after the post-treatment nadir PSA is achieved. Traditional hormonal therapy is the mainstay of systemic treatment for PSA-only recurrence, although nontraditional approaches such as intermittent and oral-only hormonal therapy are under study. PMID- 11062384 TI - Abarelix Depot, a GnRH antagonist, v LHRH superagonists in prostate cancer: differential effects on follicle-stimulating hormone. Abarelix Depot study group. AB - PURPOSE: A Phase II clinical study contrasted the endocrinologic and biochemical efficacy of Abarelix Depot, a gonadotropin-releasing hormone (GnRH) antagonist, with luteinizing hormone releasing-hormone (LHRH) superagonists, with or without additional antiandrogens, in men with prostate cancer. METHODS: This study was open-label and treated 242 men. Abarelix Depot 100 mg was administered by intramuscular injection to 209 men, and LHRH, with or without an antiandrogen, was administered to 33 men according to the formulation used. Serum concentrations of follicle-stimulating hormone (FSH) and other hormones were measured at baseline and at specified time points for the first 85 days of the study. Median serum concentrations of FSH at baseline were similar for the two treatment groups. RESULTS: Men treated with LHRH superagonists, with or without an antiandrogen, had a surge in the serum concentration of FSH on day 2 before FSH concentrations started to decline. Men in the Abarelix Depot group had an immediate and sustained decrease in the serum concentration of FSH. CONCLUSION: Recent data suggest that FSH may be an independent growth factor for prostate cancer. The Abarelix Depot-induced decreased in FSH may have a role in the treatment of men with endocrine- responsive disease or for those men whose disease has escaped from hormone sensitivity. PMID- 11062385 TI - Urinary incontinence after treatment for localized prostate cancer. AB - The incidence of incontinence after radical prostatectomy has ranged from 0 to 57% depending on the series and the type of incontinence considered. When total incontinence (not minimal stress incontinence) is reported, the average incidence is no more than 5%. This figure will increase with age, and in most series, approximately 10% of patients around the age of 70 will have total incontinence postoperatively. Preservation of continence after radical prostatectomy depends largely on the preservation of the distal urethral smooth-muscle sphincteric mechanism, which begins at the pelvic floor and ends at the prostatourethral junction. Newer techniques that attempt to increase postoperative continence include not cutting the puboprostatic ligaments and attempting to preserve as much striated muscle as possible along the length of the remaining urethra. Patients who are incontinent for 6 months after the surgery with no evidence of improvement will probably not become continent on their own. Therefore, some type of therapy should be considered. The options are periurethral injection of a bulking agent, implantation of an artificial sphincter, and, most recently, a bulbourethral sling procedure. PMID- 11062386 TI - UsToo PC-SPES surveys: review of studies and update of previous survey results. AB - In 1997, we resolved to survey UsToo members and other men known at that time to be taking PC-SPES, a Chinese herb combination that contains eight herbs: chrysanthemum, dyers woad, licorice, reishi, san-qi ginseng, rabdosia, saw palmetto, and baikal skullcap. The survey showed positive results, with respondents experiencing a decline in serum prostate specific antigen (PSA), most to the undetectable range. Of these patients, 88% maintained a low PSA concentration, whereas 12% had a rise from nadir. These results made it obvious that we should obtain follow-up reports from the respondents. We therefore conducted a second survey, this time finding 93% of the respondents with positive results and only 7% reporting a rise in PSA after the initial lowering with PC SPES. Even though there are some side effects, a great majority of men are realizing good PSA control while taking the capsules, and some of the respondents are now into their fourth year of PC-SPES use. Currently, several institutions are investigating the biology of this Chinese herb combination. Although there is some estrogenic effect, there are other potential mechanisms of action to enable this product to control PSA, not only in newly diagnosed cancer, but also in longer-term use. PMID- 11062387 TI - Left in the dust of the campaign trail. PMID- 11062388 TI - The small world of metabolism. PMID- 11062389 TI - Good faith gone bad. PMID- 11062391 TI - Errata PMID- 11062390 TI - Bioweapons protocol update. PMID- 11062392 TI - Power shortages affect firms PMID- 11062393 TI - Clinical trials halted PMID- 11062395 TI - GMO roundup PMID- 11062394 TI - UK allows genetic testing PMID- 11062396 TI - AAAS cautions against germ line gene therapy PMID- 11062398 TI - Biotech fundraising, Q3 2000 V. Q2 2000 PMID- 11062397 TI - Structural genomics boost PMID- 11062399 TI - GM food policy upheld PMID- 11062400 TI - Affymetrix spins off firm PMID- 11062401 TI - Research collaborations PMID- 11062402 TI - New xeno joint venture PMID- 11062403 TI - Cloning for conservation PMID- 11062404 TI - Differentiating ES cells PMID- 11062405 TI - New twist on fluorescent antibodies PMID- 11062406 TI - Meningococcal mutagenesis PMID- 11062409 TI - Strategy for clean gene transformation PMID- 11062408 TI - Wavelength shifters PMID- 11062407 TI - Gene detection by array PMID- 11062411 TI - An edible hepatitis vaccine PMID- 11062410 TI - Crop storage without all that rot PMID- 11062412 TI - Antibody gene vaccine tested in fish? PMID- 11062413 TI - Gene delivery in angioplasty PMID- 11062414 TI - A transgene control system PMID- 11062416 TI - Technical reports PMID- 11062415 TI - TNF homes in on tumor vessels PMID- 11062419 TI - Profiling plant metabolites PMID- 11062417 TI - Review PMID- 11062418 TI - Keeping IL-2 toxicity at bay PMID- 11062420 TI - rFactor VIII deficit questioned. PMID- 11062421 TI - Research suggests importance of haplotypes over SNPs. PMID- 11062422 TI - Estonian parliament considers genome law. PMID- 11062423 TI - Taco dispute underscores need for standardized tests. PMID- 11062424 TI - Gene-therapy death prompts broad civil lawsuit. PMID- 11062425 TI - Italian GMO ban could spread. PMID- 11062426 TI - MMPI demise spotlights target choice. PMID- 11062427 TI - Genes, greens, and vaccines. PMID- 11062428 TI - Metabolic profiling on the right path. PMID- 11062429 TI - Three little pigs worth the huff and puff? PMID- 11062430 TI - Fishing for vaccines. PMID- 11062431 TI - The challenges of in silico biology. PMID- 11062432 TI - Transgenic plants as factories for biopharmaceuticals. AB - Plants have considerable potential for the production of biopharmaceutical proteins and peptides because they are easily transformed and provide a cheap source of protein. Several biotechnology companies are now actively developing, field testing, and patenting plant expression systems, while clinical trials are proceeding on the first biopharmaceuticals derived from them. One transgenic plant-derived biopharmaceutical, hirudin, is now being commercially produced in Canada for the first time. Product purification is potentially an expensive process, and various methods are currently being developed to overcome this problem, including oleosin-fusion technology, which allows extraction with oil bodies. In some cases, delivery of a biopharmaceutical product by direct ingestion of the modified plant potentially removes the need for purification. Such biopharmaceuticals and edible vaccines can be stored and distributed as seeds, tubers, or fruits, making immunization programs in developing countries cheaper and potentially easier to administer. Some of the most expensive biopharmaceuticals of restricted availability, such as glucocerebrosidase, could become much cheaper and more plentiful through production in transgenic plants. PMID- 11062433 TI - Metabolite profiling for plant functional genomics. AB - Multiparallel analyses of mRNA and proteins are central to today's functional genomics initiatives. We describe here the use of metabolite profiling as a new tool for a comparative display of gene function. It has the potential not only to provide deeper insight into complex regulatory processes but also to determine phenotype directly. Using gas chromatography/mass spectrometry (GC/MS), we automatically quantified 326 distinct compounds from Arabidopsis thaliana leaf extracts. It was possible to assign a chemical structure to approximately half of these compounds. Comparison of four Arabidopsis genotypes (two homozygous ecotypes and a mutant of each ecotype) showed that each genotype possesses a distinct metabolic profile. Data mining tools such as principal component analysis enabled the assignment of "metabolic phenotypes" using these large data sets. The metabolic phenotypes of the two ecotypes were more divergent than were the metabolic phenotypes of the single-loci mutant and their parental ecotypes. These results demonstrate the use of metabolite profiling as a tool to significantly extend and enhance the power of existing functional genomics approaches. PMID- 11062434 TI - Transgenic plants expressing cationic peptide chimeras exhibit broad-spectrum resistance to phytopathogens. AB - Here we describe a strategy for engineering transgenic plants with broad-spectrum resistance to bacterial and fungal phytopathogens. We expressed a synthetic gene encoding a N terminus-modified, cecropin-melittin cationic peptide chimera (MsrA1), with broad-spectrum antimicrobial activity. The synthetic gene was introduced into two potato (Solanum tuberosum L.) cultivars, Desiree and Russet Burbank, stable incorporation was confirmed by PCR and DNA sequencing, and expression confirmed by reverse transcription (RT)-PCR and recovery of the biologically active peptide. The morphology and yield of transgenic Desiree plants and tubers was unaffected. Highly stringent challenges with bacterial or fungal phytopathogens demonstrated powerful resistance. Tubers retained their resistance to infectious challenge for more than a year, and did not appear to be harmful when fed to mice. Expression of msrA1 in the cultivar Russet Burbank caused a striking lesion-mimic phenotype during leaf and tuber development, indicating its utility may be cultivar specific. Given the ubiquity of antimicrobial cationic peptides as well as their inherent capacity for recombinant and combinatorial variants, this approach may potentially be used to engineer a range of disease-resistant plants. PMID- 11062435 TI - Production of hepatitis B surface antigen in transgenic plants for oral immunization. AB - Here we present data showing oral immunogenicity of recombinant hepatitis B surface antigen (HBsAg) in preclinical animal trials. Mice fed transgenic HBsAg potato tubers showed a primary immune response (increases in HBsAg-specific serum antibody) that could be greatly boosted by intraperitoneal delivery of a single subimmunogenic dose of commercial HBsAg vaccine, indicating that plants expressing HBsAg in edible tissues may be a new means for oral hepatitis B immunization. However, attainment of such a goal will require higher HBsAg expression than was observed for the potatoes used in this study. We conducted a systematic analysis of factors influencing the accumulation of HBsAg in transgenic potato, including 5' and 3' flanking elements and protein targeting within plant cells. The most striking improvements resulted from (1) alternative polyadenylation signals, and (2) fusion proteins containing targeting signals designed to enhance integration or retention of HBsAg in the endoplasmic reticulum (ER) of plant cells. PMID- 11062436 TI - Removal of antibiotic resistance genes from transgenic tobacco plastids. AB - Removal of antibiotic resistance genes from genetically modified (GM) crops removes the risk of their transfer to the environment or gut microbes. Integration of foreign genes into plastid DNA enhances containment in crops that inherit their plastids maternally. Efficient plastid transformation requires the aadA marker gene, which confers resistance to the antibiotics spectinomycin and streptomycin. We have exploited plastid DNA recombination and cytoplasmic sorting to remove aadA from transplastomic tobacco plants. A 4.9 kbp insert, composed of aadA flanked by bar and uidA genes, was integrated into plastid DNA and selected to remove wild-type plastid genomes. The bar gene confers tolerance to the herbicide glufosinate despite being GC-rich. Excision of aadA and uidA mediated by two 174 bp direct repeats generated aadA-free T(0) transplastomic plants containing the bar gene. Removal of aadA and bar by three 418 bp direct repeats allowed the isolation of marker-free T(2) plants containing a plastid-located uidA reporter gene. PMID- 11062437 TI - Immunoprophylaxis in fish by injection of mouse antibody genes. AB - Antibodies are a crucial part of the body's specific defense against infectious diseases and have considerable potential as therapeutic and prophylactic agents in humans and animals. The development of recombinant single-chain antibodies allows a genetic application strategy for prevention of infectious diseases. To test this in a fish model, a gene construct encoding a neutralizing single-chain antibody to the fish-pathogenic rhabdovirus VHSV (viral hemorrhagic septicemia virus) was administered to rainbow trout by intramuscular injection of plasmid DNA. Circulating recombinant antibodies could later be detected in the fish, and protective immunity to the viral disease was established. PMID- 11062438 TI - Gene delivery from a DNA controlled-release stent in porcine coronary arteries. AB - Expandable intra-arterial stents are widely used for treating coronary disease. We hypothesized that local gene delivery could be achieved with the controlled release of DNA from a polymer coating on an expandable stent. Our paper reports the first successful transfection in vivo using a DNA controlled-release stent. Green fluorescent protein (GFP) plasmid DNA within emulsion-coated stents was efficiently expressed in cell cultures (7.9% +/- 0.7% vs. 0.6% +/- 0.2% control, p < 0.001) of rat aortic smooth muscle cells. In a series of pig stent angioplasty studies, GFP expression was observed in all coronary arteries (normal, nondiseased) in the DNA-treated group, but not in control arteries. GFP plasmid DNA in the arterial wall was confirmed by PCR, and GFP presence in the pig coronaries was confirmed by immunohistochemistry. Thus, DNA-eluting stents are capable of arterial transfection, and could be useful as delivery systems for candidate vectors for gene therapy of cardiovascular diseases. PMID- 11062439 TI - Enhancement of tumor necrosis factor alpha antitumor immunotherapeutic properties by targeted delivery to aminopeptidase N (CD13). AB - The clinical use of tumor necrosis factor alpha (TNF) as an anticancer drug is limited to local treatments because of its dose-limiting systemic toxicity. We show here that murine TNF fused with CNGRC peptide (NGR-TNF), an aminopeptidase N (CD13) ligand that targets activated blood vessels in tumors, is 12-15 times more efficient than murine TNF in decreasing the tumor burden in lymphoma and melanoma animal models, whereas its toxicity is similar. Similarly, human NGR-TNF induced stronger antitumor effects than human TNF, even with 30 times lower doses. Coadministration of murine NGR-TNF with a CNGRC peptide or an anti-CD13 antibody markedly decreased its antitumor effects. Tumor regression, induced by doses of murine NGR-TNF lower than the LD50, was accompanied by protective immunity. In contrast, no cure was induced by TNF at any dose. These results suggest that targeted delivery of TNF to CD13 may enhance its immunotherapeutic properties. Moreover, these findings reveal the potential of tumor homing peptides to generate a new class of recombinant cytokines that compared to immunocytokines have a simpler structure, could be easier to produce and are potentially less immunogenic. PMID- 11062440 TI - Wavelength-shifting molecular beacons. AB - We describe wavelength-shifting molecular beacons, which are nucleic acid hybridization probes that fluoresce in a variety of different colors, yet are excited by a common monochromatic light source. The twin functions of absorption of energy from the excitation light and emission of that energy in the form of fluorescent light are assigned to two separate fluorophores in the same probe. These probes contain a harvester fluorophore that absorbs strongly in the wavelength range of the monochromatic light source, an emitter fluorophore of the desired emission color, and a nonfluorescent quencher. In the absence of complementary nucleic acid targets, the probes are dark, whereas in the presence of targets, they fluoresce-not in the emission range of the harvester fluorophore that absorbs the light, but rather in the emission range of the emitter fluorophore. This shift in emission spectrum is due to the transfer of the absorbed energy from the harvester fluorophore to the emitter fluorophore by fluorescence resonance energy transfer, and it only takes place in probes that are bound to targets. Wavelength-shifting molecular beacons are substantially brighter than conventional molecular beacons that contain a fluorophore that cannot efficiently absorb energy from the available monochromatic light source. We describe the spectral characteristics of wavelength-shifting molecular beacons, and we demonstrate how their use improves and simplifies multiplex genetic analyses. PMID- 11062441 TI - A T-cell-selective interleukin 2 mutein exhibits potent antitumor activity and is well tolerated in vivo. AB - Human interleukin 2 (IL-2; Proleukin) is an approved therapeutic for advanced stage metastatic cancer; however, its use is restricted because of severe systemic toxicity. Its function as a central mediator of T-cell activation may contribute to its efficacy for cancer therapy. However, activation of natural killer (NK) cells by therapeutically administered IL-2 may mediate toxicity. Here we have used targeted mutagenesis of human IL-2 to generate a mutein with approximately 3,000-fold in vitro selectivity for T cells over NK cells relative to wild-type IL-2. We compared the variant, termed BAY 50-4798, with human IL-2 (Proleukin) in a therapeutic dosing regimen in chimpanzees, and found that although the T-cell mobilization and activation properties of BAY 50-4798 were comparable to human IL-2, BAY 50-4798 was better tolerated in the chimpanzee. BAY 50-4798 was also shown to inhibit metastasis in a mouse tumor model. These results indicate that BAY 50-4798 may exhibit a greater therapeutic index than IL 2 in humans in the treatment of cancer and AIDS. PMID- 11062443 TI - Research errata PMID- 11062442 TI - Streptogramin-based gene regulation systems for mammalian cells. AB - Here we describe repressible (PipOFF) as well as inducible (PipON) systems for regulated gene expression in mammalian cells, based on the repressor Pip (pristinamycin-induced protein), which is encoded by the streptogramin resistance operon of Streptomyces coelicolor. Expression of genes placed under control of these systems was responsive to clinically approved antibiotics belonging to the streptogramin group (pristinamycin, virginiamycin, and Synercid). The versatility of these systems was demonstrated by streptogramin-regulated expression of mouse erythropoietin (EPO), human placental secreted alkaline phosphatase (SEAP), or green fluorescent protein (GFP) in diverse cell lines (BHK, CHO, HeLa, and mouse myoblasts). Analysis of isogenic constructs in CHO cells demonstrated the PipOFF system gave lower background and higher induction ratios than the widely used tetracycline-repressible (TetOFF) expression systems. The streptogramin-based expression technology was functionally compatible with the TetOFF system, thus enabling the selective use of different antibiotics to independently control two different gene activities in the same cell. PMID- 11062444 TI - A hyperthermostable bacterial histone-like protein as an efficient mediator for transfection of eukaryotic cells. AB - Gene delivery has shown potential in a variety of applications, including basic research, therapies for inborn genetic defects, cancer, AIDS, tissue engineering, and vaccination. Most available systems have serious drawbacks, such as safety hazards, inefficiency under in vivo-like conditions, and expensive production. When using naked DNA, for instance, a large amount of ultrapure DNA has to be applied as a result of degradation by nucleases. Similarly, the use of eukaryotic histones, synthetic peptides, or peptide nucleic acids may be limited by high production costs. We have demonstrated a biotechnologically feasible and economical approach for gene delivery using the histone-like protein from the hyperthermostable eubacterium Thermotoga maritima, TmHU as an efficient gene transfer reagent. HU can be easily isolated from recombinant Escherichia coli, is extraordinarily stable, and protects dsDNA from thermal denaturation. This study demonstrates its use as an inexpensive tool for gene delivery. PMID- 11062445 TI - Rapid genotyping by MALDI-monitored nuclease selection from probe libraries. AB - Data on five single-nucleotide polymorphisms (SNPs) per gene are estimated to allow association of disease risks or pharmacogenetic parameters with individual genes. Efficient technologies for rapidly detecting SNPs will therefore facilitate the mining of genomic information. Known methods for SNP analysis include restriction-fragment-length polymorphism polymerase chain reaction (PCR), allele-specific oligomer hybridization, oligomer-specific ligation assays, minisequencing, direct sequencing, fluorescence-detected 5'-exonuclease assays, and hybridization with PNA probes. Detection by mass spectrometry (MS) offers speed and high resolution. Matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI TOF MS) can detect primer extension products, mass-tagged oligonucleotides, DNA created by restriction endonuclease cleavage, and genomic DNA. We have previously reported MALDI-TOF-monitored nuclease selections of modified oligonucleotides with increased affinity for targets. Here we use nuclease selections for genotyping by treating DNA to be analyzed with oligonucleotide probes representing known genotypes and digesting probes that are not complementary to the DNA. With phosphodiesterase I, the target-bound, complementary probe is largely refractory to nuclease attack and its peak persists in mass spectra (Fig. 1A). In optimized assays, both alleles of a heterozygote were genotyped with six nonamer DNA probes (> or = 125 fmol each) and asymmetrically amplified DNA from exon 10 of the cystic fibrosis transmembrane regulatory gene (CFTR). PMID- 11062446 TI - Finding common cause in the patent debate. PMID- 11062447 TI - Recent patents in xenotransplantation PMID- 11062449 TI - New products PMID- 11062448 TI - People PMID- 11062450 TI - Testing times. PMID- 11062451 TI - The past within us. PMID- 11062452 TI - Of giant axons and curly hair. PMID- 11062453 TI - Viva la revolution! A report from the FANTOM meeting. PMID- 11062454 TI - When more is better. PMID- 11062456 TI - TOUCHINGbase PMID- 11062455 TI - A new face of the Rhesus antigen. PMID- 11062457 TI - Behaviour and the standardization fallacy. PMID- 11062458 TI - Dominant isolated renal magnesium loss is caused by misrouting of the Na(+),K(+) ATPase gamma-subunit. AB - Primary hypomagnesaemia is composed of a heterogeneous group of disorders characterized by renal or intestinal Mg(2+) wasting, often associated with disturbances in Ca(2+) excretion. We identified a putative dominant-negative mutation in the gene encoding the Na(+), K(+)-ATPase gamma-subunit (FXYD2), leading to defective routing of the protein in a family with dominant renal hypomagnesaemia. PMID- 11062459 TI - Mater, a maternal effect gene required for early embryonic development in mice. AB - Maternal effect genes produce mRNA or proteins that accumulate in the egg during oogenesis. We show here that Mater, a mouse oocyte protein dependent on the maternal genome, is essential for embryonic development beyond the two-cell stage. Females lacking the maternal effect gene Mater are sterile. Null males are fertile. PMID- 11062460 TI - Mutations in SDHC cause autosomal dominant paraganglioma, type 3. AB - Nonchromaffin paragangliomas (PGLs) are usually benign, neural-crest-derived, slow-growing tumours of parasympathetic ganglia. Between 10% and 50% of cases are familial and are transmitted as autosomal dominant traits with incomplete and age dependent penetrance. PMID- 11062461 TI - Mutations in MERTK, the human orthologue of the RCS rat retinal dystrophy gene, cause retinitis pigmentosa. AB - Mutation of a receptor tyrosine kinase gene, Mertk, in the Royal College of Surgeons (RCS) rat results in defective phagocytosis of photoreceptor outer segments by the retinal pigment epithelium (RPE) and retinal degeneration. We screened the human orthologue, MERTK, located at 2q14.1 (ref. 10), in 328 DNA samples from individuals with various retinal dystrophies and found three mutations in three individuals with retinitis pigmentosa (RP). Our findings are the first conclusive evidence implicating the RPE phagocytosis pathway in human retinal disease. PMID- 11062462 TI - Normal telomere lengths found in cloned cattle. AB - Success of cloning using adult somatic cells has been reported in sheep, mice and cattle. The report that 'Dolly' the sheep, the first clone from an adult mammal, inherited shortened telomeres from her cell donor and that her telomeres were further shortened by the brief culture of donor cells has raised serious scientific and public concerns about the 'genetic age' and potential developmental problems of cloned animals. This observation was challenged by a recent report that showed calves cloned from fetal cells have longer telomeres than their age-matched controls. The question remains whether Dolly's short telomeres were an exception or a general fact, which would differ from the telomeres of fetal-derived clones. PMID- 11062463 TI - Mutations in the gene encoding the latency-associated peptide of TGF-beta 1 cause Camurati-Engelmann disease. AB - Camurati-Engelmann disease (CED; MIM 131300), or progressive diaphyseal dysplasia, is a rare, sclerosing bone dysplasia inherited in an autosomal dominant manner. Recently, the gene causing CED has been assigned to the chromosomal region 19q13 (refs 1-3). Because this region contains the gene encoding transforming growth factor-beta 1 (TGFB1), an important mediator of bone remodelling, we evaluated TGFB1 as a candidate gene for causing CED. PMID- 11062464 TI - The nicotinic receptor beta 2 subunit is mutant in nocturnal frontal lobe epilepsy. AB - Clustered attacks of epileptic episodes originating from the frontal lobe during sleep are the main symptoms of autosomal dominant nocturnal frontal lobe epilepsy (ADNFLE, MIM 600513). Despite the clinical homogeneity, three forms of ADNFLE have been associated with chromosomes 20 (ENFL1; ref. 1), 15 (ENFL2; ref. 2) and 1 (ENFL3; ref. 3). Mutations of the gene encoding the neuronal nicotinic acetylcholine receptor alpha 4 subunit (CHRNA4 ) have been found in ADNFLE-ENFL1 families, but these mutations account for only a small proportion of ADNFLE cases. The newly identified locus associated with ENFL3 harbours several candidate genes, including CHRNB2 (ref. 8), whose gene product, the beta 2 nicotinic acetylcholine receptor (nAChR) subunit, co-assembles with the alpha 4 nAChR subunit to form the active receptor. PMID- 11062465 TI - Genes and addiction. AB - Drug addiction, like all psychiatric disorders, is defined solely in behavioural terms. For example, addiction can be considered a loss of control over drug taking, or compulsive drug-seeking and -taking despite horrendous consequences. Abnormal behaviours are a consequence of aberrant brain function, which means that it is a tangible goal to identify the biological underpinnings of addiction. The genetic basis of addiction encompasses two broad areas of enquiry. One of these is the identification of genetic variation in humans that partly determines susceptibility to addiction. The other is the use of animal models to investigate the role of specific genes in mediating the development of addiction. Whereas recent advances in this latter effort are heartening, a major challenge remains: to understand how the many genes implicated in rodent models interact to yield as complex a phenotype as addiction. PMID- 11062466 TI - Analysis of yeast protein kinases using protein chips. AB - We have developed a novel protein chip technology that allows the high-throughput analysis of biochemical activities, and used this approach to analyse nearly all of the protein kinases from Saccharomyces cerevisiae. Protein chips are disposable arrays of microwells in silicone elastomer sheets placed on top of microscope slides. The high density and small size of the wells allows for high throughput batch processing and simultaneous analysis of many individual samples. Only small amounts of protein are required. Of 122 known and predicted yeast protein kinases, 119 were overexpressed and analysed using 17 different substrates and protein chips. We found many novel activities and that a large number of protein kinases are capable of phosphorylating tyrosine. The tyrosine phosphorylating enzymes often share common amino acid residues that lie near the catalytic region. Thus, our study identified a number of novel features of protein kinases and demonstrates that protein chip technology is useful for high throughput screening of protein biochemical activity. PMID- 11062467 TI - Senescence bypass screen identifies TBX2, which represses Cdkn2a (p19(ARF)) and is amplified in a subset of human breast cancers. AB - To identify new immortalizing genes with potential roles in tumorigenesis, we performed a genetic screen aimed to bypass the rapid and tight senescence arrest of primary fibroblasts deficient for the oncogene Bmi1. We identified the T-box member TBX2 as a potent immortalizing gene that acts by downregulating Cdkn2a (p19(ARF)). TBX2 represses the Cdkn2a (p19(ARF)) promoter and attenuates E2F1, Myc or HRAS-mediated induction of Cdkn2a (p19(ARF)). We found TBX2 to be amplified in a subset of primary human breast cancers, indicating that it might contribute to breast cancer development. PMID- 11062468 TI - Inactivation of Hdh in the brain and testis results in progressive neurodegeneration and sterility in mice. AB - Inactivation of the mouse homologue of the Huntington disease gene (Hdh) results in early embryonic lethality. To investigate the normal function of Hdh in the adult and to evaluate current models for Huntington disease (HD), we have used the Cre/loxP site-specific recombination strategy to inactivate Hdh expression in the forebrain and testis, resulting in a progressive degenerative neuronal phenotype and sterility. On the basis of these results, we propose that huntingtin is required for neuronal function and survival in the brain and that a loss-of-function mechanism may contribute to HD pathogenesis. PMID- 11062469 TI - Sublimiting concentration of TFIIH transcription/DNA repair factor causes TTD-A trichothiodystrophy disorder. AB - The repair-deficient form of trichothiodystrophy (TTD) most often results from mutations in the genes XPB or XPD, encoding helicases of the transcription/repair factor TFIIH. The genetic defect in a third group, TTD-A, is unknown, but is also caused by dysfunctioning TFIIH. None of the TFIIH subunits carry a mutation and TFIIH from TTD-A cells is active in both transcription and repair. Instead, immunoblot and immunofluorescence analyses reveal a strong reduction in the TFIIH concentration. Thus, the phenotype of TTD-A appears to result from sublimiting amounts of TFIIH, probably due to a mutation in a gene determining the complex stability. The reduction of TFIIH mainly affects its repair function and hardly influences transcription. PMID- 11062470 TI - Mining the human genome using microarrays of open reading frames. AB - To test the hypothesis that the human genome project will uncover many genes not previously discovered by sequencing of expressed sequence tags (ESTs), we designed and produced a set of microarrays using probes based on open reading frames (ORFs) in 350 Mb of finished and draft human sequence. Our approach aims to identify all genes directly from genomic sequence by querying gene expression. We analysed genomic sequence with a suite of ORF prediction programs, selected approximately one ORF per gene, amplified the ORFs from genomic DNA and arrayed the amplicons onto treated glass slides. Of the first 10,000 arrayed ORFs, 31% are completely novel and 29% are similar, but not identical, to sequences in public databases. Approximately one-half of these are expressed in the tissues we queried by microarray. Subsequent verification by other techniques confirmed expression of several of the novel genes. Expressed sequence tags (ESTs) have yielded vast amounts of data, but our results indicate that many genes in the human genome will only be found by genomic sequencing. PMID- 11062471 TI - Mutations in NYX, encoding the leucine-rich proteoglycan nyctalopin, cause X linked complete congenital stationary night blindness. AB - During development, visual photoreceptors, bipolar cells and other neurons establish connections within the retina enabling the eye to process visual images over approximately 7 log units of illumination. Within the retina, cells that respond to light increment and light decrement are separated into ON- and OFF pathways. Hereditary diseases are known to disturb these retinal pathways, causing either progressive degeneration or stationary deficits. Congenital stationary night blindness (CSNB) is a group of stable retinal disorders that are characterized by abnormal night vision. Genetic subtypes of CSNB have been defined and different disease actions have been postulated. The molecular bases have been elucidated in several subtypes, providing a better understanding of the disease mechanisms and developmental retinal neurobiology. Here we have studied 22 families with 'complete' X-linked CSNB (CSNB1; MIM 310500; ref. 4) in which affected males have night blindness, some photopic vision loss and a defect of the ON-pathway. We have found 14 different mutations, including 1 founder mutation in 7 families from the United States, in a novel candidate gene, NYX. NYX, which encodes a glycosylphosphatidyl (GPI)-anchored protein called nyctalopin, is a new and unique member of the small leucine-rich proteoglycan (SLRP) family. The role of other SLRP proteins suggests that mutant nyctalopin disrupts developing retinal interconnections involving the ON-bipolar cells, leading to the visual losses seen in patients with complete CSNB. PMID- 11062472 TI - The complete form of X-linked congenital stationary night blindness is caused by mutations in a gene encoding a leucine-rich repeat protein. AB - X-linked congenital stationary night blindness (XLCSNB) is characterized by impaired scotopic vision with associated ocular symptoms such as myopia, hyperopia, nystagmus and reduced visual acuity. Genetic mapping in families with XLCSNB revealed two different loci on the proximal short arm of the X chromosome. These two genetic subtypes can be distinguished on the basis of electroretinogram (ERG) responses and psychophysical testing as a complete (CSNB1) and an incomplete (CSNB2) form. The CSNB1 locus has been mapped to a 5-cM linkage interval in Xp11.4 (refs 2,5-7). Here we construct and analyse a contig between the markers DXS993 and DXS228, leading to the identification of a new gene mutated in CSNB1 patients. It is partially deleted in 3 families and mutation analysis in a further 21 families detected another 13 different mutations. This gene, designated NYX, encodes a protein of 481 amino acids (nyctalopin) and is expressed at low levels in tissues including retina, brain, testis and muscle. The predicted polypeptide is a glycosylphosphatidylinositol (GPI)-anchored extracellular protein with 11 typical and 2 cysteine-rich, leucine-rich repeats (LRRs). This motif is important for protein-protein interactions and members of the LRR superfamily are involved in cell adhesion and axon guidance. Future functional analysis of nyctalopin might therefore give insight into the fine regulation of cell-cell contacts in the retina. PMID- 11062473 TI - Arteriovenous malformations in mice lacking activin receptor-like kinase-1. AB - The mature circulatory system is comprised of two parallel, yet distinct, vascular networks that carry blood to and from the heart. Studies have suggested that endothelial tubes are specified as arteries and veins at the earliest stages of angiogenesis, before the onset of circulation. To understand the molecular basis for arterial-venous identity, we have focused our studies on a human vascular dysplasia, hereditary haemorrhagic telangiectasia (HHT), wherein arterial and venous beds fail to remain distinct. Genetic studies have demonstrated that HHT can be caused by loss-of-function mutations in the gene encoding activin receptor-like kinase-1 (ACVRL1; ref. 5). ACVRL1 encodes a type I receptor for the TGF-beta superfamily of growth factors. At the earliest stage of vascular development, mice lacking Acvrl1 develop large shunts between arteries and veins, downregulate arterial Efnb2 and fail to confine intravascular haematopoiesis to arteries. These mice die by mid-gestation with severe arteriovenous malformations resulting from fusion of major arteries and veins. The early loss of anatomical, molecular and functional distinctions between arteries and veins indicates that Acvrl1 is required for developing distinct arterial and venous vascular beds. PMID- 11062474 TI - Mutant WD-repeat protein in triple-A syndrome. AB - Triple-A syndrome (MIM 231550; also known as Allgrove syndrome) is an autosomal recessive disorder characterized by adrenocorticotropin hormone (ACTH)-resistant adrenal insufficiency, achalasia of the oesophageal cardia and alacrima. Whereas several lines of evidence indicate that triple-A syndrome results from the abnormal development of the autonomic nervous system, late-onset progressive neurological symptoms (including cerebellar ataxia, peripheral neuropathy and mild dementia) suggest that the central nervous system may be involved in the disease as well. Using fine-mapping based on linkage disequilibrium in North African inbred families, we identified a short ancestral haplotype on chromosome 12q13 (<1 cM), sequenced a BAC contig encompassing the triple-A minimal region and identified a novel gene (AAAS) encoding a protein of 547 amino acids that is mutant in affected individuals. We found five homozygous truncating mutations in unrelated patients and ascribed the founder effect in North African families to a single splice-donor site mutation that occurred more than 2,400 years ago. The predicted product of AAAS, ALADIN (for alacrima-achalasia-adrenal insufficiency neurologic disorder), belongs to the WD-repeat family of regulatory proteins, indicating a new disease mechanism involved in triple-A syndrome. The expression of the gene in both neuroendocrine and cerebral structures points to a role in the normal development of the peripheral and central nervous systems. PMID- 11062475 TI - Impaired insulin secretion and beta-cell loss in tissue-specific knockout mice with mitochondrial diabetes. AB - Mitochondrial dysfunction is an important contributor to human pathology and it is estimated that mutations of mitochondrial DNA (mtDNA) cause approximately 0.5 1% of all types of diabetes mellitus. We have generated a mouse model for mitochondrial diabetes by tissue-specific disruption of the nuclear gene encoding mitochondrial transcription factor A (Tfam, previously mtTFA; ref. 7) in pancreatic beta-cells. This transcriptional activator is imported to mitochondria, where it is essential for mtDNA expression and maintenance. The Tfam-mutant mice developed diabetes from the age of approximately 5 weeks and displayed severe mtDNA depletion, deficient oxidative phosphorylation and abnormal appearing mitochondria in islets at the ages of 7-9 weeks. We performed physiological studies of beta-cell stimulus-secretion coupling in islets isolated from 7-9-week-old mutant mice and found reduced hyperpolarization of the mitochondrial membrane potential, impaired Ca(2+)-signalling and lowered insulin release in response to glucose stimulation. We observed reduced beta-cell mass in older mutants. Our findings identify two phases in the pathogenesis of mitochondrial diabetes; mutant beta-cells initially display reduced stimulus secretion coupling, later followed by beta-cell loss. This animal model reproduces the beta-cell pathology of human mitochondrial diabetes and provides genetic evidence for a critical role of the respiratory chain in insulin secretion. PMID- 11062476 TI - The human Rhesus-associated RhAG protein and a kidney homologue promote ammonium transport in yeast. AB - The Rhesus blood-group antigens are defined by a complex association of membrane polypeptides that includes the non-glycosylated Rh proteins (RhD and RhCE) and the RHag glycoprotein, which is strictly required for cell surface expression of these antigens. RhAG and the Rh polypeptides are erythroid-specific transmembrane proteins belonging to the same family (36% identity). Despite their importance in transfusion medicine, the function of RhAG and Rh proteins remains unknown, except that their absence in Rh(null) individuals leads to morphological and functional abnormalities of erythrocytes, known as the Rh-deficiency syndrome. We recently found significant sequence similarity between the Rh family proteins, especially RhAG, and Mep/Amt ammonium transporters. We show here that RhAG and also RhGK, a new human homologue expressed in kidney cells only, function as ammonium transport proteins when expressed in yeast. Both specifically complement the growth defect of a yeast mutant deficient in ammonium uptake. Moreover, ammonium efflux assays and growth tests in the presence of toxic concentrations of the analogue methylammonium indicate that RhAG and RhGK also promote ammonium export. Our results provide the first experimental evidence for a direct role of RhAG and RhGK in ammonium transport. These findings are of high interest, because no specific ammonium transport system has been characterized so far in human. PMID- 11062477 TI - Autosomal dominant hypophosphataemic rickets is associated with mutations in FGF23. AB - Proper serum phosphate concentrations are maintained by a complex and poorly understood process. Identification of genes responsible for inherited disorders involving disturbances in phosphate homeostasis may provide insight into the pathways that regulate phosphate balance. Several hereditary disorders of isolated phosphate wasting have been described, including X-linked hypophosphataemic rickets (XLH), hypophosphataemic bone disease (HBD), hereditary hypophosphataemic rickets with hypercalciuria (HHRH) and autosomal dominant hypophosphataemic rickets (ADHR). Inactivating mutations of the gene PHEX, encoding a member of the neutral endopeptidase family of proteins, are responsible for XLH (refs 6,7). ADHR (MIM 193100) is characterized by low serum phosphorus concentrations, rickets, osteomalacia, lower extremity deformities, short stature, bone pain and dental abscesses. Here we describe a positional cloning approach used to identify the ADHR gene which included the annotation of 37 genes within 4 Mb of genomic sequence. We identified missense mutations in a gene encoding a new member of the fibroblast growth factor (FGF) family, FGF23. These mutations in patients with ADHR represent the first mutations found in a human FGF gene. PMID- 11062478 TI - Histone deacetylase interacts directly with DNA topoisomerase II. AB - Histone deacetylases (HDACs) modify nucleosomal histones, have a key role in the regulation of gene transcription, and may be involved in cell-cycle regulation, differentiation and human cancer. Purified recombinant human HDAC1 protein was used to screen a cDNA expression library, and one of the clones identified encoded DNA topoisomerase II (Topo II), an enzyme known to have a role in transcriptional regulation and chromatin organization. Coimmunoprecipitation experiments indicate that HDAC1 and HDAC2 are associated with Topo II in vivo under normal physiological conditions. Complexes containing Topo II possess HDAC activities, and complexes containing HDAC1 or HDAC2 possess Topo II activities. HDAC and Topo II modify each other's activity in vitro and in vivo. Our results indicate the existence of a functionally coupled complex between these two enzymes and offer insights into the potential mechanisms of action of both enzymes. PMID- 11062479 TI - IgA nephropathy, the most common cause of glomerulonephritis, is linked to 6q22 23. AB - End-stage renal disease (ESRD) is a major public health problem, affecting 1 in 1,000 individuals and with an annual death rate of 20% despite dialysis treatment. IgA nephropathy (IgAN) is the most common form of glomerulonephritis, a principal cause of ESRD worldwide; it affects up to 1.3% of the population and its pathogenesis is unknown. Kidneys of people with IgAN show deposits of IgA containing immune complexes with proliferation of the glomerular mesangium (Fig. 1). Typical clinical features include onset before age 40 with haematuria and proteinuria (blood and protein in the urine), and episodes of gross haematuria following mucosal infections are common; 30% of patients develop progressive renal failure. Although not generally considered a hereditary disease, striking ethnic variation in prevalence and familial clustering, along with subclinical renal abnormalities among relatives of IgAN cases, have suggested a heretofore undefined genetic component. By genome-wide analysis of linkage in 30 multiplex IgAN kindreds, we demonstrate linkage of IgAN to 6q22-23 under a dominant model of transmission with incomplete penetrance, with a lod score of 5.6 and 60% of kindreds linked. These findings for the first time indicate the existence of a locus with large effect on development of IgAN and identify the chromosomal location of this disease gene. PMID- 11062480 TI - Y chromosome sequence variation and the history of human populations. AB - Binary polymorphisms associated with the non-recombining region of the human Y chromosome (NRY) preserve the paternal genetic legacy of our species that has persisted to the present, permitting inference of human evolution, population affinity and demographic history. We used denaturing high-performance liquid chromatography (DHPLC; ref. 2) to identify 160 of the 166 bi-allelic and 1 tri allelic site that formed a parsimonious genealogy of 116 haplotypes, several of which display distinct population affinities based on the analysis of 1062 globally representative individuals. A minority of contemporary East Africans and Khoisan represent the descendants of the most ancestral patrilineages of anatomically modern humans that left Africa between 35,000 and 89,000 years ago. PMID- 11062481 TI - A common variant in BRCA2 is associated with both breast cancer risk and prenatal viability. AB - Inherited mutations in the gene BRCA2 predispose carriers to early onset breast cancer, but such mutations account for fewer than 2% of all cases in East Anglia. It is likely that low penetrance alleles explain the greater part of inherited susceptibility to breast cancer; polymorphic variants in strongly predisposing genes, such as BRCA2, are candidates for this role. BRCA2 is thought to be involved in DNA double strand break-repair. Few mice in which Brca2 is truncated survive to birth; of those that do, most are male, smaller than their normal littermates and have high cancer incidence. Here we show that a common human polymorphism (N372H) in exon 10 of BRCA2 confers an increased risk of breast cancer: the HH homozygotes have a 1.31-fold (95% CI, 1.07-1.61) greater risk than the NN group. Moreover, in normal female controls of all ages there is a significant deficiency of homozygotes compared with that expected from Hardy Weinberg equilibrium, whereas in males there is an excess of homozygotes: the HH group has an estimated fitness of 0.82 in females and 1.38 in males. Therefore, this variant of BRCA2 appears also to affect fetal survival in a sex-dependent manner. PMID- 11062482 TI - Loss-of-function mutations in the EGF-CFC gene CFC1 are associated with human left-right laterality defects. AB - All vertebrates display a characteristic asymmetry of internal organs with the cardiac apex, stomach and spleen towards the left, and the liver and gall bladder on the right. Left-right (L-R) axis abnormalities or laterality defects are common in humans (1 in 8,500 live births). Several genes (such as Nodal, Ebaf and Pitx2) have been implicated in L-R organ positioning in model organisms. In humans, relatively few genes have been associated with a small percentage of human situs defects. These include ZIC3 (ref. 5), LEFTB (formerly LEFTY2; ref. 6) and ACVR2B (encoding activin receptor IIB; ref. 7). The EGF-CFC genes, mouse Cfc1 (encoding the Cryptic protein; ref. 9) and zebrafish one-eyed pinhead (oep; refs 10, 11) are essential for the establishment of the L-R axis. EGF-CFC proteins act as co-factors for Nodal-related signals, which have also been implicated in L-R axis development. Here we identify loss-of-function mutations in human CFC1 (encoding the CRYPTIC protein) in patients with heterotaxic phenotypes (randomized organ positioning). The mutant proteins have aberrant cellular localization in transfected cells and are functionally defective in a zebrafish oep-mutant rescue assay. Our findings indicate that the essential role of EGF-CFC genes and Nodal signalling in left-right axis formation is conserved from fish to humans. Moreover, our results support a role for environmental and/or genetic modifiers in determining the ultimate phenotype in humans. PMID- 11062483 TI - The gene encoding gigaxonin, a new member of the cytoskeletal BTB/kelch repeat family, is mutated in giant axonal neuropathy. AB - Disorganization of the neurofilament network is a prominent feature of several neurodegenerative disorders including amyotrophic lateral sclerosis (ALS), infantile spinal muscular atrophy and axonal Charcot-Marie-Tooth disease. Giant axonal neuropathy (GAN, MIM 256850), a severe, autosomal recessive sensorimotor neuropathy affecting both the peripheral nerves and the central nervous system, is characterized by neurofilament accumulation, leading to segmental distension of the axons. GAN corresponds to a generalized disorganization of the cytoskeletal intermediate filaments (IFs), to which neurofilaments belong, as abnormal aggregation of multiple tissue-specific IFs has been reported: vimentin in endothelial cells, Schwann cells and cultured skin fibroblasts, and glial fibrillary acidic protein (GFAP) in astrocytes. Keratin IFs also seem to be alterated, as most patients present characteristic curly or kinky hairs. We report here identification of the gene GAN, which encodes a novel, ubiquitously expressed protein we have named gigaxonin. We found one frameshift, four nonsense and nine missense mutations in GAN of GAN patients. Gigaxonin is composed of an amino-terminal BTB (for Broad-Complex, Tramtrack and Bric a brac) domain followed by a six kelch repeats, which are predicted to adopt a beta-propeller shape. Distantly related proteins sharing a similar domain organization have various functions associated with the cytoskeleton, predicting that gigaxonin is a novel and distinct cytoskeletal protein that may represent a general pathological target for other neurodegenerative disorders with alterations in the neurofilament network. PMID- 11062484 TI - Proliferating cell nuclear antigen and Msh2p-Msh6p interact to form an active mispair recognition complex. AB - Proliferating cell nuclear antigen (PCNA) is required for mismatch repair (MMR) and has been shown to interact with complexes containing Msh2p or MLH1 (refs 1 4). PCNA has been implicated to act in MMR before and during the DNA synthesis step, although the biochemical basis for the role of PCNA early in MMR is unclear. Here we observe an interaction between PCNA and Msh2p-Msh6p mediated by a specific PCNA-binding site present in Msh6p. An msh6 mutation that eliminated the PCNA-binding site caused a mutator phenotype and a defect in the interaction with PCNA. The association of PCNA with Msh2p-Msh6p stimulated the preferential binding of Msh2p-Msh6p to DNA containing mispaired bases. Mutant PCNA proteins encoded by MMR-defective pol30 alleles were defective for interaction with Msh2p Msh6p and for stimulation of mispair binding by Msh2p-Msh6p. Our results suggest that PCNA functions directly in mispair recognition and that mispair recognition requires a higher-order complex containing proteins in addition to Msh2p-Msh6p. PMID- 11062486 TI - Corrections/Erratum PMID- 11062485 TI - Hypoglycaemia, liver necrosis and perinatal death in mice lacking all isoforms of phosphoinositide 3-kinase p85 alpha. AB - Phosphoinositide 3-kinases produce 3'-phosphorylated phosphoinositides that act as second messengers to recruit other signalling proteins to the membrane. Pi3ks are activated by many extracellular stimuli and have been implicated in a variety of cellular responses. The Pi3k gene family is complex and the physiological roles of different classes and isoforms are not clear. The gene Pik3r1 encodes three proteins (p85 alpha, p55 alpha and p50 alpha) that serve as regulatory subunits of class IA Pi3ks (ref. 2). Mice lacking only the p85 alpha isoform are viable but display hypoglycaemia and increased insulin sensitivity correlating with upregulation of the p55 alpha and p50 alpha variants. Here we report that loss of all protein products of Pik3r1 results in perinatal lethality. We observed, among other abnormalities, extensive hepatocyte necrosis and chylous ascites. We also noted enlarged skeletal muscle fibres, brown fat necrosis and calcification of cardiac tissue. In liver and muscle, loss of the major regulatory isoform caused a great decrease in expression and activity of class IA Pi3k catalytic subunits; nevertheless, homozygous mice still displayed hypoglycaemia, lower insulin levels and increased glucose tolerance. Our findings reveal that p55 alpha and/or p50 alpha are required for survival, but not for development of hypoglycaemia, in mice lacking p85 alpha. PMID- 11062487 TI - Faring of animal research. PMID- 11062488 TI - Drosophila, the savvy exterminator. PMID- 11062489 TI - Immunotherapy of human cancer: lessons from mice. PMID- 11062490 TI - Cytokines, virokines and the evolution of immunity. PMID- 11062491 TI - Understanding how pre-B cells come of age. PMID- 11062492 TI - Fc alpha/microR: single member or first born in the family? PMID- 11062493 TI - Signaling for cytotoxicity. PMID- 11062495 TI - Immunology highlights from the recent literature. PMID- 11062494 TI - Remembrance of things past. PMID- 11062496 TI - Antibody regulation of B cell development. AB - Antibodies on the surface of B lymphocytes trigger adaptive immune responses and control a series of antigen-independent checkpoints during B cell development. These physiologic processes are regulated by a complex of membrane immunoglobulin and two signal transducing proteins known as Ig alpha and Ig beta. Here we focus on the role of antibodies in governing the maturation of B cells from early antigen-independent through the final antigen-dependent stages. PMID- 11062497 TI - Uncoupling of inflammatory chemokine receptors by IL-10: generation of functional decoys. AB - As originally demonstrated for the interleukin 1 (IL-1) type II receptor, some primary proinflammatory cytokines from the IL-1 and tumor necrosis factor families are regulated by decoy receptors that are structurally incapable of signaling. Here we report that concomitant exposure to proinflammatory signals and IL-10 generates functional decoy receptors in the chemokine system. Inflammatory signals, which cause dendritic cell (DC) maturation and migration to lymphoid organs, induce a chemokine receptor switch, with down-regulation of inflammatory receptors (such as CCR1, CCR2, CCR5) and induction of CCR7. Concomitant exposure to lipopolysaccharide (LPS) and IL-10 blocks the chemokine receptor switch associated with DC maturation. LPS + IL-10-treated DCs showed low expression of CCR7 and high expression of CCR1, CCR2 and CCR5. These receptors were unable to elicit migration. We provide evidence that uncoupled receptors, expressed on LPS + IL-10-treated cells, sequester and scavenge inflammatory chemokines. Similar results were obtained for monocytes exposed to activating signals and IL-10. Thus, in an inflammatory environment, IL-10 generates functional decoy receptors on DC and monocytes, which act as molecular sinks and scavengers for inflammatory chemokines. PMID- 11062498 TI - Hemokinin is a hematopoietic-specific tachykinin that regulates B lymphopoiesis. AB - We report here the molecular cloning of a newly identified preprotachykinin gene, Pptc, which specifies the sequence for a new preprotachykinin protein and bioactive peptide designated hemokinin 1 (HK-1). PPT-C mRNA was detected primarily in hematopoietic cells in contrast to the previously described Ppta and Pptb genes, which are predominantly expressed in neuronal tissues. HK-1 has several biological activities that are similar to the most studied tachykinin, substance P, such as induction of plasma extravasation and mast cell degranulation. However, HK-1 also has properties that are indicative of a critical role in mouse B cell development. HK-1 stimulated the proliferation of interleukin 7-expanded B cell precursors, whereas substance P had no effect. HK 1, but not substance P, promoted the survival of freshly isolated bone marrow B lineage cells or cultured, lipopolysaccharide-stimulated pre-B cells. N-acetyl-L trytophan-3,5-bistrifluromethyl benzyl ester, a tachykinin receptor antagonist, increased apoptosis of these cells and in vivo administration of this antagonist led to specific reductions of the B220lowCD43 population (the pre-B cell compartment) in the bone marrow and the IgMhighIgDlow population (the newly generated B cells) in the spleen. Thus, HK-1 may be an autocrine factor that is important for the survival of B cell precursors at a critical phase of development. PMID- 11062499 TI - Pattern recognition receptors TLR4 and CD14 mediate response to respiratory syncytial virus. AB - The innate immune system contributes to the earliest phase of the host defense against foreign organisms and has both soluble and cellular pattern recognition receptors for microbial products. Two important members of this receptor group, CD14 and the Toll-like receptor (TLR) pattern recognition receptors, are essential for the innate immune response to components of Gram-negative and Gram positive bacteria, mycobacteria, spirochetes and yeast. We now find that these receptors function in an antiviral response as well. The innate immune response to the fusion protein of an important respiratory pathogen of humans, respiratory syncytial virus (RSV), was mediated by TLR4 and CD14. RSV persisted longer in the lungs of infected TLR4-deficient mice compared to normal mice. Thus, a common receptor activation pathway can initiate innate immune responses to both bacterial and viral pathogens. PMID- 11062500 TI - Regulating T helper cell immunity through antigen responsiveness and calcium entry. AB - We evaluated changes in the signaling potentials and proliferative capacity of single antigen-specific T helper (TH) cells during a primary immune response to a protein antigen. At the peak of cellular expansion in vivo all antigen-specific TH cells exhibited a profound block in CD3- and CD4-mediated mobilization of intracellular calcium together with a more global block in T cell receptor independent capacitative calcium entry (CCE). The proliferative response of these antigen-specific TH cells to anti-CD3, anti-CD28 and IL-2 was also severely blunted. Cross-linking CD69 on a substantial fraction of CD69+ antigen-specific TH cells relieved this block in CCE and restored proliferative capacity in vitro. The CCE rescue operated through a CD69-coupled G protein and required calcium bound calmodulin and calcineurin. These data reveal critical changes in the responsiveness of antigen-specific TH cells and provide evidence of new mechanisms for the regulation of antigen-specific TH cell development in vivo. PMID- 11062501 TI - Two new proteases in the MHC class I processing pathway. AB - The proteasome generates exact major histocompatibility complex (MHC) class I ligands as well as NH2-terminal-extended precursor peptides. The proteases responsible for the final NH2-terminal trimming of the precursor peptides had, until now, not been determined. By using specific selective criteria we purified two cytosolic proteolytic activities, puromycin-sensitive aminopeptidase and bleomycin hydrolase. These proteases could remove NH2-terminal amino acids from the vesicular stomatitis virus nucleoprotein cytotoxic T cell epitope 52-59 (RGYVYQGL) resulting, in combination with proteasomes, in the generation of the correct epitope. Our data provide evidence for the existence of redundant systems acting downstream of the proteasome in the antigen-processing pathway for MHC class I molecules. PMID- 11062502 TI - Pivotal role of phosphoinositide-3 kinase in regulation of cytotoxicity in natural killer cells. AB - The mitogen-activated protein kinase-extracellular signal-regulated kinase signaling element (MAPK-ERK) plays a critical role in natural killer (NK) cell lysis of tumor cells, but its upstream effectors were previously unknown. We show that inhibition of phosphoinositide-3 kinase (PI3K) in NK cells blocks p21 activated kinase 1 (PAK1), MAPK kinase (MEK) and ERK activation by target cell ligation, interferes with perforin and granzyme B movement toward target cells and suppresses NK cytotoxicity. Dominant-negative N17Rac1 and PAK1 mimic the suppressive effects of PI3K inhibitors, whereas constitutively active V12Rac1 has the opposite effect. V12Rac1 restores the activity of downstream effectors and lytic function in LY294002- or wortmannin-treated, but not PD98059-treated, NK cells. These results document a specific PI3K-->Rac1-->PAK1-->MEK-->ERK pathway in NK cells that effects lysis. PMID- 11062503 TI - Interleukin-7 mediates the homeostasis of naive and memory CD8 T cells in vivo. AB - The naive and memory T lymphocyte pools are maintained through poorly understood homeostatic mechanisms that may include signaling via cytokine receptors. We show that interleukin-7 (IL-7) plays multiple roles in regulating homeostasis of CD8+ T cells. We found that IL-7 was required for homeostatic expansion of naive CD8+ and CD4+ T cells in lymphopenic hosts and for CD8+ T cell survival in normal hosts. In contrast, IL-7 was not necessary for growth of CD8+ T cells in response to a virus infection but was critical for generating T cell memory. Up-regulation of Bcl-2 in the absence of IL-7 signaling was impaired after activation in vivo. Homeostatic proliferation of memory cells was also partially dependent on IL-7. These results point to IL-7 as a pivotal cytokine in T cell homeostasis. PMID- 11062504 TI - CD27 is required for generation and long-term maintenance of T cell immunity. AB - The Traf-linked tumor necrosis factor receptor family member CD27 is known as a T cell costimulatory molecule. We generated CD27-/- mice and found that CD27 makes essential contributions to mature CD4+ and CD8+ T cell function: CD27 supported antigen-specific expansion (but not effector cell maturation) of naive T cells, independent of the cell cycle-promoting activities of CD28 and interleukin 2. Primary CD4+ and CD8+ T cell responses to influenza virus were impaired in CD27-/ mice. Effects of deleting the gene encoding CD27 were most profound on T cell memory, reflected by delayed response kinetics and reduction of CD8+ virus specific T cell numbers to the level seen in the primary response. This demonstrates the requirement for a costimulatory receptor in the generation of T cell memory. PMID- 11062506 TI - Crystal structure of a T cell receptor bound to an allogenic MHC molecule. PMID- 11062505 TI - Fc alpha/mu receptor mediates endocytosis of IgM-coated microbes. AB - IgM is the first antibody to be produced in a humoral immune response and plays an important role in the primary stages of immunity. Here we describe a mouse Fc receptor, designated Fc alpha/microR, and its human homolog, that bind both IgM and IgA with intermediate or high affinity. Fc alpha/microR is constitutively expressed on the majority of B lymphocytes and macrophages. Cross-linking Fc alpha/microR expressed on a pro-B cell line Ba/F3 transfectant with soluble IgM or IgM-coated microparticles induced internalization of the receptor. Fc alpha/microR also mediated primary B lymphocyte endocytosis of IgM-coated Staphylococcus aureus. Thus, Fc alpha/microR is involved in the primary stages of the immune response to microbes. PMID- 11062507 TI - Clinical trials and tribulations. PMID- 11062508 TI - The QUEST trial, a paradigm of HIV collaborative research. PMID- 11062509 TI - Merger signals shift in xenotransplantation research. PMID- 11062510 TI - Skepticism surrounds triple Nobel winners. PMID- 11062511 TI - Abortion drug approval opens up research PMID- 11062512 TI - NIH gives money for orphan development. PMID- 11062513 TI - USDA gives ground over lab animals. PMID- 11062514 TI - Placebo trials deemed unethical. PMID- 11062515 TI - Brazilian studies supported long-term PMID- 11062516 TI - South African village prepares for first HIV vaccine trial. PMID- 11062517 TI - UK insurers allowed to use genetic tests. PMID- 11062518 TI - . . . While government causes confusion. PMID- 11062520 TI - Ethical issues in biomedical publication PMID- 11062519 TI - The National Institutes of Health and clinical research: a progress report. PMID- 11062521 TI - The triple helix: gene, organism and environment PMID- 11062522 TI - Parkinson disease gene therapy moves toward the clinic. PMID- 11062523 TI - Normal and mutant huntingtin: partners in crime. PMID- 11062525 TI - Seeing is believing PMID- 11062524 TI - To be or notch to be. PMID- 11062526 TI - Stem cell alchemy. PMID- 11062527 TI - Split stories converge at desmoglein 1. PMID- 11062528 TI - Meningococcal genomics: two steps forward, one step back. PMID- 11062529 TI - Not a minute to waste. PMID- 11062530 TI - Malaria vaccines-targeting infected hepatocytes. PMID- 11062531 TI - Aids vaccine advance. PMID- 11062532 TI - Decoding calcium signals involved in cardiac growth and function. AB - Calcium is central in the regulation of cardiac contractility, growth and gene expression. Variations in the amplitude, frequency and compartmentalization of calcium signals are decoded by calcium/calmodulin-dependent enzymes, ion channels and transcription factors. Understanding the circuitry for calcium signaling creates opportunities for pharmacological modification of cardiac function. PMID- 11062533 TI - Purified hematopoietic stem cells can differentiate into hepatocytes in vivo. AB - The characterization of hepatic progenitor cells is of great scientific and clinical interest. Here we report that intravenous injection of adult bone marrow cells in the FAH(-/-) mouse, an animal model of tyrosinemia type I, rescued the mouse and restored the biochemical function of its liver. Moreover, within bone marrow, only rigorously purified hematopoietic stem cells gave rise to donor derived hematopoietic and hepatic regeneration. This result seems to contradict the conventional assumptions of the germ layer origins of tissues such as the liver, and raises the question of whether the cells of the hematopoietic stem cell phenotype are pluripotent hematopoietic cells that retain the ability to transdifferentiate, or whether they are more primitive multipotent cells. PMID- 11062534 TI - Stem cell repopulation efficiency but not pool size is governed by p27(kip1). AB - Sustained blood cell production requires preservation of a quiescent, multipotential stem cell pool that intermittently gives rise to progenitors with robust proliferative potential. The ability of cells to shift from a highly constrained to a vigorously active proliferative state is critical for maintaining stem cells while providing the responsiveness necessary for host defense. The cyclin-dependent kinase inhibitor (CDKI), p21(cip1/waf1) (p21) dominates stem cell kinetics. Here we report that another CDKI, p27(kip1) (p27), does not affect stem cell number, cell cycling, or self-renewal, but markedly alters progenitor proliferation and pool size. Therefore, distinct CDKIs govern the highly divergent stem and progenitor cell populations. When competitively transplanted, p27-deficient stem cells generate progenitors that eventually dominate blood cell production. Modulating p27 expression in a small number of stem cells may translate into effects on the majority of mature cells, thereby providing a strategy for potentiating the impact of transduced cells in stem cell gene therapy. PMID- 11062535 TI - Deficiency in caspase-9 or caspase-3 induces compensatory caspase activation. AB - Dysregulation of apoptosis contributes to the pathogenesis of many human diseases. As effectors of the apoptotic machinery, caspases are considered potential therapeutic targets. Using an established in vivo model of Fas-mediated apoptosis, we demonstrate here that elimination of certain caspases was compensated in vivo by the activation of other caspases. Hepatocyte apoptosis and mouse death induced by the Fas agonistic antibody Jo2 required proapoptotic Bcl-2 family member Bid and used a Bid-mediated mitochondrial pathway of caspase activation; deficiency in caspases essential for this pathway, caspase-9 or caspase-3, unexpectedly resulted in rapid activation of alternate caspases after injection of Jo2, and therefore failed to protect mice against Jo2 toxicity. Moreover, both ultraviolet and gamma irradiation, two established inducers of the mitochondrial caspase-activation pathway, also elicited compensatory activation of caspases in cultured caspase-3(-/-) hepatocytes, indicating that the compensatory caspase activation was mediated through the mitochondria. Our findings provide direct experimental evidence for compensatory pathways of caspase activation. This issue should therefore be considered in developing caspase inhibitors for therapeutic applications. PMID- 11062536 TI - A peptide-doxorubicin 'prodrug' activated by prostate-specific antigen selectively kills prostate tumor cells positive for prostate-specific antigen in vivo. AB - We covalently linked doxorubicin with a peptide that is hydrolyzable by prostate specific antigen. In the presence of prostate tumor cells secreting prostate specific antigen, the peptide moiety of this conjugate, L-377,202, was hydrolyzed, resulting in the release of leucine-doxorubicin and doxorubicin, which are both very cytotoxic to cancer cells. However, L-377,202 was much less cytotoxic than conventional doxorubicin to cells in culture that do not secrete prostate-specific antigen. L-377,202 was approximately 15 times more effective than was conventional doxorubicin at inhibiting the growth of human prostate cancer tumors in nude mice when both drugs were used at their maximally tolerated doses. Nude mice inoculated with human prostate tumor cells secreting prostate specific antigen showed considerable reductions in tumor burden with minimal total body weight loss when treated with L-377, 202. This improvement in therapeutic index correlated with the selective localization of leucine doxorubicin and free doxorubicin in tissues secreting prostate-specific antigen after exposure to L-377,202. PMID- 11062537 TI - Conjugation of arginine oligomers to cyclosporin A facilitates topical delivery and inhibition of inflammation. AB - Many systemically effective drugs such as cyclosporin A are ineffective topically because of their poor penetration into skin. To surmount this problem, we conjugated a heptamer of arginine to cyclosporin A through a pH-sensitive linker to produce R7-CsA. In contrast to unmodified cyclosporin A, which fails to penetrate skin, topically applied R7-CsA was efficiently transported into cells in mouse and human skin. R7-CsA reached dermal T lymphocytes and inhibited cutaneous inflammation. These data establish a general strategy for enhancing delivery of poorly absorbed drugs across tissue barriers and provide a new topical approach to the treatment of inflammatory skin disorders. PMID- 11062538 TI - Protection against Plasmodium falciparum malaria in chimpanzees by immunization with the conserved pre-erythrocytic liver-stage antigen 3. AB - In humans, sterile immunity against malaria can be consistently induced through exposure to the bites of thousands of irradiated infected mosquitoes. The same level of protection has yet to be achieved using subunit vaccines. Recent studies have indicated an essential function for intrahepatic parasites, the stage after the mosquito bite, and thus for antigens expressed during this stage. We report here the identification of liver-stage antigen 3, which is expressed both in the mosquito and liver-stage parasites. This Plasmodium falciparum 200-kilodalton protein is highly conserved, and showed promising antigenic and immunogenic properties. In chimpanzees (Pan troglodytes), the primates most closely related to humans and that share a similar susceptibility to P. falciparum liver-stage infection, immunization with LSA-3 induced protection against successive heterologous challenges with large numbers of P. falciparum sporozoites. PMID- 11062539 TI - Cytoadhesion of Plasmodium falciparum ring-stage-infected erythrocytes. AB - A common pathological characteristic of Plasmodium falciparum infection is the cytoadhesion of mature-stage-infected erythrocytes (IE) to host endothelium and syncytiotrophoblasts. Massive accumulation of IE in the brain microvasculature or placenta is strongly correlated with severe forms of malaria. Extensive binding of IE to placental chondroitin sulfate A (CSA) is associated with physiopathology during pregnancy. The adhesive phenotype of IE correlates with the appearance of Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) at the erythrocyte surface (approximately 16 h after merozoite invasion), so that only early blood stage (ring-stage) IE appear in the peripheral blood. Here, we describe results that challenge the existing view of blood-stage IE biology by demonstrating the specific adhesion of IE, during the early ring-stage, to endothelial cell lines from the brain and lung and to placental syncytiotrophoblasts. Later, during blood-stage development of these IE, trophozoites switch to an exclusively CSA cytoadhesion phenotype. Therefore, adhesion to an individual endothelial cell or syncytiotrophoblast may occur throughout the blood-stage cycle, indicating the presence in malaria patients of noncirculating (cryptic) parasite subpopulations. We detected two previously unknown parasite proteins on the surface of ring-stage IE. These proteins disappear shortly after the start of PfEMP1-mediated adhesion. PMID- 11062540 TI - Functional genomics of Neisseria meningitidis pathogenesis. AB - The pathogenic bacterium Neisseria meningitidis is an important cause of septicemia and meningitis, especially in childhood. The establishment and maintenance of bacteremic infection is a pre-requisite for all the pathological sequelae of meningococcal infection. To further understand the genetic basis of this essential step in pathogenesis, we analyzed a library of 2,850 insertional mutants of N. meningitidis for their capacity to cause systemic infection in an infant rat model. The library was constructed by in vitro modification of Neisseria genomic DNA with the purified components of Tn10 transposition. We identified 73 genes in the N. meningitidis genome that are essential for bacteremic disease. Eight insertions were in genes encoding known pathogenicity factors. Involvement of the remaining 65 genes in meningocoocal pathogenesis has not been demonstrated previously, and the identification of these genes provides insights into the pathogenic mechanisms that underlie meningococcal infection. Our results provide a genome-wide analysis of the attributes of N. meningitidis required for disseminated infection, and may lead to new interventions to prevent and treat meningococcal infection. PMID- 11062541 TI - Toxin in bullous impetigo and staphylococcal scalded-skin syndrome targets desmoglein 1. AB - Exfoliative toxin A, produced by Staphylococcus aureus, causes blisters in bullous impetigo and its more generalized form, staphylococcal scalded-skin syndrome. The toxin shows exquisite specificity in causing loss of cell adhesion only in the superficial epidermis. Although exfoliative toxin A has the structure of a serine protease, a target protein has not been identified. Desmoglein (Dsg) 1, a desmosomal cadherin that mediates cell-cell adhesion, may be the target of exfoliative toxin A, because it is the target of autoantibodies in pemphigus foliaceus, in which blisters form with identical tissue specificity and histology. We show here that exfoliative toxin A cleaved mouse and human Dsg1, but not closely related cadherins such as Dsg3. We demonstrate this specific cleavage in cell culture, in neonatal mouse skin and with recombinant Dsg1, and conclude that Dsg1 is the specific receptor for exfoliative toxin A cleavage. This unique proteolytic attack on the desmosome causes a blister just below the stratum corneum, which forms the epidermal barrier, presumably allowing the bacteria in bullous impetigo to proliferate and spread beneath this barrier. PMID- 11062542 TI - Pluripotent, cytokine-dependent, hematopoietic stem cells are immortalized by constitutive Notch1 signaling. AB - Hematopoietic stem cells give rise to progeny that either self-renew in an undifferentiated state or lose self-renewal capabilities and commit to lymphoid or myeloid lineages. Here we evaluated whether hematopoietic stem cell self renewal is affected by the Notch pathway. Notch signaling controls cell fate choices in both invertebrates and vertebrates by inhibiting certain differentiation pathways, thereby permitting cells to either differentiate along an alternative pathway or to self-renew. Notch receptors are present in hematopoietic precursors and Notch signaling enhances the in vitro generation of human and mouse hematopoietic precursors, determines T- or B-cell lineage specification from a common lymphoid precursor and promotes expansion of CD8(+) cells. Here, we demonstrate that constitutive Notch1 signaling in hematopoietic cells established immortalized, cytokine-dependent cell lines that generated progeny with either lymphoid or myeloid characteristics both in vitro and in vivo. These data support a role for Notch signaling in regulating hematopoietic stem cell self-renewal. Furthermore, the establishment of clonal, pluripotent cell lines provides the opportunity to assess mechanisms regulating stem cell commitment and demonstrates a general method for immortalizing stem cell populations for further analysis. PMID- 11062543 TI - Human mesenchymal stem cells engraft and demonstrate site-specific differentiation after in utero transplantation in sheep. AB - Mesenchymal stem cells are multipotent cells that can be isolated from adult bone marrow and can be induced in vitro and in vivo to differentiate into a variety of mesenchymal tissues, including bone, cartilage, tendon, fat, bone marrow stroma, and muscle. Despite their potential clinical utility for cellular and gene therapy, the fate of mesenchymal stem cells after systemic administration is mostly unknown. To address this, we transplanted a well-characterized human mesenchymal stem cell population into fetal sheep early in gestation, before and after the expected development of immunologic competence. In this xenogeneic system, human mesenchymal stem cells engrafted and persisted in multiple tissues for as long as 13 months after transplantation. Transplanted human cells underwent site-specific differentiation into chondrocytes, adipocytes, myocytes and cardiomyocytes, bone marrow stromal cells and thymic stroma. Unexpectedly, there was long-term engraftment even when cells were transplanted after the expected development of immunocompetence. Thus, mesenchymal stem cells maintain their multipotential capacity after transplantation, and seem to have unique immunologic characteristics that allow persistence in a xenogeneic environment. Our data support the possibility of the transplantability of mesenchymal stem cells and their potential utility in tissue engineering, and cellular and gene therapy applications. PMID- 11062544 TI - A new approach for analyzing proton magnetic resonance spectroscopic images of brain tumors: nosologic images. PMID- 11062546 TI - On the market PMID- 11062545 TI - Human immunodeficiency virus type 1 reverse transcriptase and protease mutation search engine for queries. PMID- 11062547 TI - Breadth plus depth. PMID- 11062548 TI - How does a bacterium find its middle? PMID- 11062549 TI - Opening doors into the proteasome. PMID- 11062550 TI - Getting to the bottom of the F1-ATPase. PMID- 11062551 TI - Catching rays. PMID- 11062552 TI - Picture story. The 'serpin'ator. PMID- 11062553 TI - Analyzing protein functions in four dimensions. AB - Time-resolved structural studies on biomolecular function are coming of age. Focus has shifted from studies on 'systems of opportunities' to a more problem oriented approach, addressing significant questions in biology and chemistry. An important step in this direction has been the use of physical and chemical trapping methods to capture and then freeze reaction intermediates in crystals. Subsequent monochromatic data collection at cryogenic temperatures can produce high resolution structures of otherwise elusive intermediates. The combination of diffraction methods with spectroscopic techniques provides a means to directly correlate electronic transitions with structural transitions in the sample, eliminating much of the guesswork from experiments. Studies on cytochrome P450, isopenicillin N synthase, cytochrome cd1 nitrite reductase, copper amine oxidase and bacteriorhodopsin were selected as examples, and the results are discussed. PMID- 11062554 TI - Structural basis for the topological specificity function of MinE. AB - Correct positioning of the division septum in Escherichia coli depends on the coordinated action of the MinC, MinD and MinE proteins. Topological specificity is conferred on the MinCD division inhibitor by MinE, which counters MinCD activity only in the vicinity of the preferred midcell division site. Here we report the structure of the homodimeric topological specificity domain of Escherichia coli MinE and show that it forms a novel alphabeta sandwich. Structure-directed mutagenesis of conserved surface residues has enabled us to identify a spatially restricted site on the surface of the protein that is critical for the topological specificity function of MinE. PMID- 11062555 TI - Molecular mechanism of NPF recognition by EH domains. AB - Eps15 homology (EH) domains are protein interaction modules that recognize Asn Pro-Phe (NPF) motifs in their biological ligands to mediate critical events during endocytosis and signal transduction. To elucidate the structural basis of the EH-NPF interaction, the solution structures of two EH-NPF complexes were solved using NMR spectroscopy. The first complex contains a peptide representing the Hrb C-terminal NPFL motif; the second contains a peptide in which an Arg residue substitutes the C-terminal Leu. The NPF residues are almost completely embedded in a hydrophobic pocket on the EH domain surface and the backbone of NPFX adopts a conformation reminiscent of the Asx-Pro type I beta-turn motif. The residue directly following NPF is crucial for recognition and is required to complete the beta-turn. Five amino acids on the EH surface mediate specific recognition of this residue through hydrophobic and electrostatic contacts. The complexes explain the selectivity of the second EH domain of Eps15 for NPF over DPF motifs and reveal a critical aromatic interaction that provides a conserved anchor for the recognition of FW, WW, SWG and HTF ligands by other EH domains. PMID- 11062556 TI - Phosphorylation of T cell receptor zeta is regulated by a lipid dependent folding transition. AB - The cytoplasmic domain of the T cell receptor zeta subunit (zeta(cyt)) is sufficient to couple receptor ligation to intracellular signaling cascades, but little is known about its structure or mechanism of signaling. In aqueous solution, zeta(cyt) is unstructured. Here we report that in the presence of lipid vesicles zeta(cyt) assumes a folded structure. The folding transition is reversible and dependent on the presence of acidic phospholipids. In the lipid bound conformation, zeta(cyt) is refractory to phosphorylation by src family tyrosine kinases, which are believed to play a key role in signal initiation in vivo. In the lipid-free, unstructured form, zeta(cyt) is readily phosphorylated, and phospho-zeta cyt exhibits neither membrane association nor structure induction. The conformational change may provide a mechanism for coupling receptor clustering to cytoplasmic signaling events. PMID- 11062557 TI - Energetic contribution of tRNA hybrid state formation to translocation catalysis on the ribosome. AB - Upon transpeptidylation, the 3' end of aminoacyl-tRNA (aa-tRNA) in the ribosomal A site enters the A/P hybrid state. We report that transpeptidylation of Phe-tRNA to fMetPhe-tRNA on Escherichia coli ribosomes substantially lowers the kinetic stability of the ribosome-tRNA complex and decreases the affinity by 18.9 kJ mol( 1). At the same time, the free energy of activation of elongation factor G dependent translocation decreases by 12.5 kJ mol(-1), indicating that part of the free energy of transpeptidylation is used to drive translocation kinetically. Thus, the formation of the A/P hybrid state constitutes an important element of the translocation mechanism. PMID- 11062558 TI - Minimal catalytic domain of a group I self-splicing intron RNA. AB - The self-splicing intron ribozymes have been regarded as primitive forms of the splicing machinery for eukaryotic pre-mRNAs. The splicing activity of group I self-splicing introns is dependent on an absolutely conserved and exceptionally densely packed core region composed of two helical domains, P3-P7 and P4-P6, that are connected rigidly via base triples. Here we show that a mutant group I intron ribozyme lacking both the P4-P6 domain and the base triples can perform the phosphoester transfer reactions required for splicing at both the 5' and 3' splice sites, demonstrating that the elements required for splicing are concentrated in the stacked helical P3-P7 domain. This finding establishes that the conserved core of the intron consists of two physically and functionally separable components, and we present a model showing the architecture of a prototype of this class of intron and the course of its molecular evolution. PMID- 11062559 TI - Germin is a manganese containing homohexamer with oxalate oxidase and superoxide dismutase activities. AB - Germin is a hydrogen peroxide generating oxalate oxidase with extreme thermal stability; it is involved in the defense against biotic and abiotic stress in plants. The structure, determined at 1.6 A resolution, comprises beta-jellyroll monomers locked into a homohexamer (a trimer of dimers), with extensive surface burial accounting for its remarkable stability. The germin dimer is structurally equivalent to the monomer of the 7S seed storage proteins (vicilins), indicating evolution from a common ancestral protein. A single manganese ion is bound per germin monomer by ligands similar to those of manganese superoxide dismutase (MnSOD). Germin is also shown to have SOD activity and we propose that the defense against extracellular superoxide radicals is an important additional role for germin and related proteins. PMID- 11062560 TI - Structure of a dioxygen reduction enzyme from Desulfovibrio gigas. AB - Desulfovibrio gigas is a strict anaerobe that contains a well-characterized metabolic pathway that enables it to survive transient contacts with oxygen. The terminal enzyme in this pathway, rubredoxin:oxygen oxidoreductase (ROO) reduces oxygen to water in a direct and safe way. The 2.5 A resolution crystal structure of ROO shows that each monomer of this homodimeric enzyme consists of a novel combination of two domains, a flavodoxin-like domain and a Zn-beta-lactamase-like domain that contains a di-iron center for dioxygen reduction. This is the first structure of a member of a superfamily of enzymes widespread in strict and facultative anaerobes, indicating its broad physiological significance. PMID- 11062561 TI - MJ0109 is an enzyme that is both an inositol monophosphatase and the 'missing' archaeal fructose-1,6-bisphosphatase. AB - In sequenced genomes, protein coding regions with unassigned function constitute between 10 and 50% of all open reading frames. Often key enzymes cannot be identified using sequence homology searches. For example, despite the fact that methanogens have an apparently functional gluconeogenesis pathway, standard tools have been unable to identify a fructose-1,6-bisphosphatase (FBPase) gene in the sequenced Methanoccocus jannaschii genome. Using a combination of functional and structural tools, we have shown that the protein product of the M. jannaschii gene MJ0109, which had been tentatively annotated as an inositol monophosphatase (IMPase), has both IMPase and FBPase activities. Moreover, several gene products annotated as IMPases from different thermophilic organisms also possess FBPase activity. Thus, we have found the FBPase that was 'missing' in thermophiles and shown that it also functions as an IMPase. PMID- 11062562 TI - Structure of the gamma-epsilon complex of ATP synthase. AB - ATP synthases (F(1)F(o)-ATPases) use energy released by the movement of protons down a transmembrane electrochemical gradient to drive the synthesis of ATP, the universal biological energy currency. Proton flow through F(o) drives rotation of a ring of c-subunits and a complex of the gamma and epsilon-subunits, causing cyclical conformational changes in F(1) that are required for catalysis. The crystal structure of a large portion of F(1) has been resolved. However, the structure of the central portion of the enzyme, through which conformational changes in F(o) are communicated to F(1), has until now remained elusive. Here we report the crystal structure of a complex of the epsilon-subunit and the central domain of the gamma-subunit refined at 2.1 A resolution. The structure reveals how rotation of these subunits causes large conformational changes in F(1), and thereby provides new insights into energy coupling between F(o) and F(1). PMID- 11062563 TI - The structure of the central stalk in bovine F(1)-ATPase at 2.4 A resolution. AB - The central stalk in ATP synthase, made of gamma, delta and epsilon subunits in the mitochondrial enzyme, is the key rotary element in the enzyme's catalytic mechanism. The gamma subunit penetrates the catalytic (alpha beta)(3) domain and protrudes beneath it, interacting with a ring of c subunits in the membrane that drives rotation of the stalk during ATP synthesis. In other crystals of F(1) ATPase, the protrusion was disordered, but with crystals of F(1)-ATPase inhibited with dicyclohexylcarbodiimide, the complete structure was revealed. The delta and epsilon subunits interact with a Rossmann fold in the gamma subunit, forming a foot. In ATP synthase, this foot interacts with the c-ring and couples the transmembrane proton motive force to catalysis in the (alpha beta)(3) domain. PMID- 11062564 TI - A gated channel into the proteasome core particle. AB - The core particle (CP) of the yeast proteasome is composed of four heptameric rings of subunits arranged in a hollow, barrel-like structure. We report that the CP is autoinhibited by the N-terminal tails of the outer (alpha) ring subunits. Crystallographic analysis showed that deletion of the tail of the alpha 3-subunit opens a channel into the proteolytically active interior chamber of the CP, thus derepressing peptide hydrolysis. In the latent state of the particle, the tails prevent substrate entry by imposing topological closure on the CP. Inhibition by the alpha-subunit tails is relieved upon binding of the regulatory particle to the CP to form the proteasome holoenzyme. PMID- 11062565 TI - Crystal structure of the multifunctional paramyxovirus hemagglutinin neuraminidase. AB - Paramyxoviruses are the main cause of respiratory disease in children. One of two viral surface glycoproteins, the hemagglutinin-neuraminidase (HN), has several functions in addition to being the major surface antigen that induces neutralizing antibodies. Here we present the crystal structures of Newcastle disease virus HN alone and in complex with either an inhibitor or with the beta anomer of sialic acid. The inhibitor complex reveals a typical neuraminidase active site within a beta-propeller fold. Comparison of the structures of the two complexes reveal differences in the active site, suggesting that the catalytic site is activated by a conformational switch. This site may provide both sialic acid binding and hydrolysis functions since there is no evidence for a second sialic acid binding site in HN. Evidence for a single site with dual functions is examined and supported by mutagenesis studies. The structure provides the basis for the structure-based design of inhibitors for a range of paramyxovirus-induced diseases. PMID- 11062566 TI - Carpal tunnel syndrome: reconciling "demand management" with clinical need. AB - Purchasers of surgical services are seeking justification for operative interventions with increasing frequency. This paper seeks to identify all relevant data currently available for carpal tunnel decompression; one of the commonest operative interventions in hand surgery. Such data, as is available, would suggest carpal tunnel decompression rates in the United Kingdom are relatively low, with fairly prolonged preoperative duration of symptoms. PMID- 11062567 TI - Relationship between the self-administered Boston questionnaire and electrophysiological findings in follow-up of surgically-treated carpal tunnel syndrome. AB - A prospective study of electrophysiological examination and the Boston self administered questionnaire (BQ) was carried out in patients with surgically treated carpal tunnel syndrome. There were 104 hands in 93 patients (13 men and 80 women, mean age 56 years). The BQ was used to assess the severity of symptoms and function, and nerve conduction studies were done before surgical release by short incision at the palm, and at follow-ups 1 and 6 months after surgery. The BQ severity score improved or became normal in 98% of hands. The mean BQ scores and distal sensory and motor conduction velocities in the median nerve showed significant improvement at the 1 month follow-up. Further significant improvement was found at 6 months. There was no relationship between the improvements in BQ scores and the distal conduction in the median nerve. The degree of improvement in sensory and motor distal conduction velocities could be forecast from presurgical values, whereas the degree of improvement in the symptoms and the functional status after release could not be predicted from the presurgical BQ scores. PMID- 11062568 TI - Is the reversed fourth dorsal metacarpal flap reliable? AB - We have carried out a study to determine if a flap based on vessels in the fourth metacarpal space could be used safely. We studied ten fresh cadaver specimens and used the flap in nine patients. In the anatomical study, we confirmed the presence of a suitable artery in nine out of the ten hands, arising from a piercing artery at the metacarpal bases, running distally under the fascia. The pivot point is located at the metacarpal heads, where the artery anastomoses to palmar branches and dorsal digital branches. In the clinical setting, the flap was reliable in eight patients. There was one case of flap necrosis. The flap seems to be reliable but several technical points are stressed to avoid complications. PMID- 11062569 TI - A comparison of dynamic extension splinting and controlled active mobilization of complete divisions of extensor tendons in zones 5 and 6. AB - We present a prospective randomized trial of two groups of 50 patients each having complete zone 5 and 6 extensor tendon injuries. These were rehabilitated by the use of either a dynamic outrigger splint or a palmar blocking splint. The results were analysed using the Miller and TAM assessments. Good and excellent results were achieved in 95 and 98% of cases following dynamic outrigger mobilization and 93 and 95% of cases using palmar blocking splint mobilization, using the Miller and TAM assessments respectively. There was no statistical difference in the results obtained between the two groups. Therefore, we prefer the latter technique which is simple, cheap, more convenient and requires less therapy time. PMID- 11062570 TI - Reduction in pain associated with open carpal tunnel decompression. AB - Patients treated by open carpal tunnel decompression under local anaesthetic experience significant pain with the introduction of local anaesthetic before operation. A prospective double-blind randomized placebo controlled trial was carried out to assess whether this pain could be reduced with the pre-application of a topical lignocaine-prilocaine anaesthetic cream. Nineteen patients undergoing simultaneous bilateral operations were studied. Pain, measured with a visual analogue scale, was significantly reduced on the sides treated with anaesthetic cream. On a four point verbal scale the rating on the placebo side was "moderate" or "severe", compared to "mild" on the anaesthetic cream side. The reduction in pain was greater in women, possibly due to a relatively thinner dermis. There were no adverse effects. PMID- 11062571 TI - Does splintage help pain after carpal tunnel release? AB - A prospective randomized single blind trial was performed of 102 patients undergoing carpal tunnel release. Patients received either a palmar plaster of Paris splint or a bulky wool and crepe bandage postoperatively for the first 48 h, to determine whether the plaster slab reduced postoperative pain. There were no statistically significant differences between the two groups in postoperative pain scores or analgesic use. PMID- 11062572 TI - Nerve stripping: new treatment for neuromas of the palmar cutaneous branch of the median nerve. AB - We present a new method for the treatment of painful neuromas of the palmar cutaneous branch of the median nerve. A preliminary cadaver study was done to investigate the extraneural and intraneural course of the palmar cutaneous branch of the median nerve with respect to the main trunk of the median nerve. Seven patients presented with a painful neuroma following previous surgery on the palmar aspect of the wrist. The neuroma was dissected and excised by stripping the whole of the palmar cutaneous branch from the main trunk of the median nerve. In all cases complete relief from pain and discomfort was achieved. The resulting area of numbness in the palm did not represent a significant problem. PMID- 11062573 TI - Centro-central union for the prevention of neuroma formation after finger amputation. AB - Sixty digits undergoing terminalization were randomized to undergo either simple transection of the digital nerves (control) or direct suture of the nerves to each other (centro-central union, CCU). Thirty-one digits were available for review with a minimum follow-up of one year following the surgery. Assessment was by visual analogue scales (VAS) and clinical examination. Measurements included static two-point discrimination (S2PD) for sensation, dolorimetry for stump tenderness and grip-strength.Resting pain, cold intolerance and perceived tenderness were comparable in the two groups. Perceived and observed touch sensitivity were better in the control group but the stumps in this group were more tender than those in the CCU group. We conclude that CCU results in more comfortable stumps but causes greater loss of sensitivity. It is a simple and worthwhile adjunct to digital amputation. PMID- 11062574 TI - The extended latissimus dorsi muscle island flap for flexion or extension of the fingers. AB - We describe the technique of transferring the latissmus dorsi muscle as an island flap for the restoration of digital flexion or extension in 28 patients. The latissmus dorsi muscle is raised down to the posterior iliac crest and prolonged with the gluteal superficial facia. We believe that this method is particularly suitable for extensive and prolonged paralysis of the lower elements of brachial plexus. It can be used also for severe Volkmann's contracture or the loss of flexor or extensor muscles of the fingers due to extensive debridement. The technique does not require microsurgery and there is no delay in reinnervation of the muscle. PMID- 11062575 TI - The prognostic value of concurrent Horner's syndrome in total obstetric brachial plexus injury. AB - The prognostic value of concurrent Horner's syndrome in infants with total birth palsy was investigated. The records of 48 cases with total palsy were reviewed. Poor spontaneous return of the motor function of the limb was found for both with and without concurrent Horner's syndrome. Fisher's exact test (P=0.02) indicated that the presence of concurrent Horner's syndrome is a significant prognostic factor for poor spontaneous recovery of the limb. PMID- 11062576 TI - Retrospective outcome analysis of staged flexor tendon reconstruction. AB - Forty staged flexor tendon reconstructions were done in 38 patients between 1991 and 1997. Results were assessed by clinical examination and questionnaire. At follow-up (mean, 35 months) a tenolysis had already been done in 12 cases. In the long fingers there was a significant difference between total active motion (187 degrees ) and total passive motion (237 degrees ). There was also a significant difference between active (24 degrees ) and passive (58 degrees ) IP motion in the thumbs. The mean power grip was 82%, pinch grip 74% and key grip 63% of the contralateral hand. None of the ten FPL reconstructions could be graded as excellent; four were good, using the criteria of Buck-Gramcko et al. (1976). Twenty-eight of the FDP reconstructions had excellent or good results. These results were better than the subjective scores given by the patients, 24 of whom complained of functional problems in daily life at follow-up. PMID- 11062577 TI - Histogenesis and morphology of the flexor tendon pulley system in the human embryonic hand. AB - The number, position, structural and ultrastructural features of the flexor tendon pulley system in six human embryonic hands, aged from 6 to 12 weeks, were studied by light and electron microscope. The pulley system can be recognized from the ninth week; later, at 12 weeks, the structures are easily identified around the flexor tendon in positions closely correlated to those found during post-natal growth and in the adult hand. Structurally and ultrastructurally the pulleys are not simply thickened portions of the sheath. They are formed by three layers: an inner layer, one or two cells thick, probably representing a parietal synovial tendon sheath; a middle layer formed by collagen bundles and fibroblasts whose direction is mainly perpendicular to the underlying phalanx; and an outermost layer consisting of mesenchymal tissue with numerous vessels which extends dorsally in an identical layer, forming a ring that includes flexor and extensor tendons and the cartilaginous model of the phalanx. The pulley does not have a semicircular shape but a much more complicated one, owing to the middle layer which in part runs dorsally and in part ventrally, under the flexor tendons. PMID- 11062578 TI - Static splinting of extensor tendon repairs. AB - In a prospective review we assessed the results of extensor tendon injuries managed postoperatively with a static splint. Thirty-three patients with 44 injured digits were assessed 4 months after primary tendon repair, using the Strickland-Glogovac criteria. Patients were managed in a static splint, the duration of which was guided by the zone of injury. Four months after repair, excellent or good results were obtained in 95%. Overall it was found that static splinting was an effective and safe method of management after extensor tendon repair. PMID- 11062579 TI - Hyaluronic acid modulates cell proliferation unequally in intrasynovial and extrasynovial rabbit tendons in vitro. AB - As tendons differ in biochemical composition and cellular capacities, we have compared dose response effects of hyaluronic acid on cell proliferation and synthesis of matrix components in intermediate and proximal segments of intrasynovial deep flexor tendons and extrasynovial peroneus rabbit tendons in vitro. Compared with matched control tendons, hyaluronic acid inhibited cell proliferation in intermediate and proximal intrasynovial flexor tendon segments at the concentrations of 0.1-2.0 mg/ml and 0.5-2.0 mg/ml respectively, but in extrasynovial tendon segments only at the concentration of 0.5 mg/ml. Hyaluronic acid did not affect synthesis of proteoglycan, collagen and non-collagen protein in either type of tendon. These results show that hyaluronic acid modulates cell proliferation unequally in intra- and extrasynovial tendons without affecting the synthesis of matrix components in the two types of tendons, indicating differential hyaluronic acid sensitivity and a possible mechanism of action. PMID- 11062580 TI - Scapholunate ligament repair using the Mitek bone anchor. AB - A retrospective study was done to assess the outcome after repair of completely ruptured scapholunate interosseous ligaments using the Mitek Mini G2 bone anchor. From 1994 to 1996. 12 patients underwent scapholunate ligament repair using the bone anchor. A follow-up assessment was done at a mean of 19 months postoperatively and revealed excellent or good results in eight patients, satisfactory in two, and poor in two patients, one of whom had developed lunate necrosis. One patient with an excellent functional result demonstrated recurrent dissociation of the scapholunate gap radiographically. The technique described proved to be simpler than conventional procedures in our hands, and yields similar functional results. PMID- 11062581 TI - Changes in hand function following wrist arthrodesis in cerebal palsy. AB - Eleven wrists in ten patients with cerebral palsy underwent wrist arthrodesis. All patients were reviewed between 6 and 121 months after surgery. Operative technique involved AO plate fixation in nine wrists. When the distal radial physis was still open (two wrists), stabilization was achieved using K-wires. A proximal row carpectomy was performed in eight patients. Soft tissue releases were necessary in three wrists. The procedure achieved its aim of improving hygiene and cosmesis. Functional improvement in the hand was noted in eight wrists. Function was consistently improved in athetoid patients. PMID- 11062582 TI - The use of the motion analysis system for evaluation of loss of movement in the finger. AB - We have used the motion analysis system to measure loss of finger movement after injury. The motion analysis system can provide information about the dynamic angular changes of each finger joint and the fingertip motion area for the injured finger. The latter can be used to calculate the percentage of fingertip motion area preserved. A stiff finger may show limited fingertip motion area with the finger joints tending to flex and extend together. PMID- 11062583 TI - A goniometric glove for clinical hand assessment. Construction, calibration and validation. AB - The construction of a goniometric glove is described. Each of the sensors in the glove was calibrated over a custom built metal hand using blocks of known angles as angular references. The digital data output from each sensor of the glove were converted into angular displacements at each joint. The glove was validated for consistency of measurement and accuracy over a custom built metal jig and in the human hand. The accuracy of the glove was found to be within the limits of traditional goniometry. It is proposed that goniometric gloves could be useful in the assessment of hand function. PMID- 11062584 TI - Treatment of chronic injuries of the ulnar collateral ligament of the thumb using a free tendon graft and bone suture anchors. AB - We report the results of a simple technique, using bone suture anchors and free tendon graft, for the reconstruction of chronic injuries of the ulnar collateral ligament complex at the thumb metacarpophalangeal (MP) joint. Our series includes 20 patients, with a mean age of 29 years. The mean follow-up period was 42 months. Using the Glickel grading system, 14 patients had excellent results and six had good results. Seventeen patients had no pain and three complained of mild pain with weather changes. Fourteen patients regained full stability of the MP joint and six had mild laxity. The mean loss of pinch strength was 18% compared with the contralateral thumb. The mean loss of motion at the metacarpophalangeal joint was 21%. PMID- 11062585 TI - Computer model analysis of the Swanson and Sutter metacarpophalangeal joint implants. AB - A representative model which mimics the behaviour of Silastic finger metacarpophalangeal joint implants was constructed using a finite element software package. The modelled implants were moved through a range of flexion, lateral deviation and a combination of both. Pistoning of both implants stems occurred within the modelled medullary cavities. For equivalent flexion angles, the Sutter implant produced a higher stress field than the Swanson implant, and the field was positioned at the central hinge mechanism. In both implants, lateral deviation increased the internal stress concentrations more than when pure flexion was applied. Overall the Swanson style of implant had lower stress magnitudes than the Sutter implant, and it is predicted that the Sutter implant will be more likely to fail than the Swanson. The failure mode for the Sutter implant would be at the central hinge region. The Swanson implant is likely to fail at the central hinge-stem interface regions. PMID- 11062586 TI - "Uneasy lies the hand in which rests the crown" an unusual foreign body following a punch injury. AB - A case of cellulitis of the hand resulting from embedding of a dental crown following a punch injury is described. This report emphasises the need for X-ray imaging in all cases of penetrating hand trauma, particularly when the history is vague, and also the difficulty in using metronidazole in alcoholic patients. PMID- 11062587 TI - Wrist arthrodesis following ulnar bar excision in Fibrodysplasia ossificans progressiva. AB - Fibrodysplasia ossificans progressiva is a rare disorder characterized by the progressive development of heterotopic bone in the connective tissues of skeletal muscle, ligaments and tendons. Surgical trauma is one of the most potent stimuli for ossification and surgical treatment is generally considered to be contraindicated in this condition. We report a good functional result in a patient with severe hand disability secondary to an ulna-carpal bar in fibrodysplasia ossificans progressiva. PMID- 11062588 TI - Isolated paralysis of the extensor pollicis longus muscle: a further variation of posterior interosseous nerve palsy. AB - A case of isolated extensor pollicis longus paralysis is reported representing a fourth, very localized, variation of posterior interosseous nerve palsy. Extensor indicis proprius transfer resulted in a return to function. As the posterior interosseous nerve supplies this and other muscles available for transfer, and the preoperative assessment of extensor indicis proprius function is difficult, the management of this localized nerve palsy presents a dilemma to the surgeon. PMID- 11062590 TI - Donor site toe morbidity should not be underestimated when considering options for thumb reconstruction. PMID- 11062592 TI - Nighttime blood pressure in elderly hypertensive patients. PMID- 11062593 TI - Preventing renal disease progression: is it the drug or the blood pressure reduction, or both? PMID- 11062594 TI - The effect of lowering blood pressure on quality of life. PMID- 11062595 TI - Hypertension and current issues in compliance and patient outcomes. AB - Economic and human costs associated with untreated or inadequately controlled hypertension and its complications continue to be an issue in the United States despite the availability of numerous antihypertensive agents. Knowledge of hypertension, product profiles, tolerability concerns, convenience of dosing, health-related quality of life effects, and cost of therapy are some of the factors that may influence the compliance of patients to their medication regimens. Recent reports on patient noncompliance have focused on patient provider relationships, psychosocial barriers, home blood pressure monitoring, and electronic monitoring systems to improve blood pressure control. The use of health-related quality of life assessment in antihypertensive studies and in routine clinical practice provides another opportunity to optimize a patient's regimen for short- and long-term hypertension control in a cost-effective manner. PMID- 11062596 TI - Observational studies of antihypertensive medication use and compliance: is drug choice a factor in treatment adherence? AB - Real-world adherence to pharmacotherapy cannot be studied in the confines of the traditional clinical trial. Thus, to better understand adherence to antihypertensive medication in actual practice, a literature search was conducted to identify observational database studies of the use of antihypertensive medication. Ten studies were identified: half studied adherence patterns after initial prescriptions to patients with a new diagnosis of hypertension, and the others evaluated antihypertensive medication use in a mix of patients with newly diagnosed or chronic hypertension. Overall, results demonstrated that adherence to treatment for hypertension in the first year is very poor. In addition, it appears that initial treatment with newer classes of drugs, such as angiotensin II antagonists, angiotensin converting enzyme inhibitors, and calcium channel blockers favors treatment adherence. This review also highlights and discusses possible under-lying factors contributing to these results and implications for physicians. PMID- 11062597 TI - Cost per millimeter of mercury lowering is a measure of economic value for antihypertensive agents. AB - The use of pharmacoeconomic analyses to evaluate the appropriateness of treatment regimens is increasing rapidly. Trials that study the efficacy of antihypertensive agents do not often measure long-term cardiovascular morbidity and mortality, so complementary methods are needed to evaluate the cost effectiveness of these agents. One method is to compare agents on the basis of their costs and blood pressure-lowering efficacy, producing a ratio of cost per millimeter of mercury lowering, or cost/mm Hg. This provides a simple, transparent method with which to evaluate the cost-effectiveness of antihypertensive agents. It also allows the cost-effectiveness of a blood pressure treatment to be assessed in terms of ambulatory blood pressure data. The use of cost/mm Hg requires assumptions that tolerability of agents is comparable and that blood pressure lowering is a valid surrogate for cardiovascular risk reduction. Given the emergence of new treatments that have differences in blood pressure efficacy, cost/mm Hg is likely to become increasingly used as an indicator of economic value. PMID- 11062598 TI - Methods and applications of quality-of-life measurement during antihypertensive therapy. AB - The measurement and evaluation of health-related quality of life during antihypertensive therapy has contributed to physicians' understanding of how patients respond to therapy in terms of physical, psychological, and social well being. Comparative clinical trials of antihypertensive drug therapies have demonstrated that treatment produces no perceptible improvements in health status. Thus, the "burden" of the therapeutic regimen from the patient's perspective is greatly magnified. Research has also demonstrated that patient expectation and preference are important components of quality-of-life assessment when the purpose of this assessment is to predict or explain human behavior. Drug properties, including mechanism of action and pharmacokinetics, have been shown to influence the impact of drug side effects and the degree to which patients tolerate therapy. Researchers have attempted to standardize the construct of quality of life to have broad applications in health care policy and management; however, clinical research in hypertension should focus on the components of health-related quality of life that influence patient outcomes with regard to behaviors affecting adherence. Antihypertensive agents that maximize patient acceptance and quality of life are now available. Future quality-of-life research should be directed toward learning more about human behavior so that adherence to nonpharmacologic interventions such as diet and exercise will be enhanced. PMID- 11062599 TI - Clinical trials report PMID- 11062600 TI - Recent advances in the management of hypertension in the elderly. AB - Important new advances have occurred in our understanding and approach to management of high blood pressure in the elderly. New clinical trials have re emphasized the risk of development of cardiovascular and cerebrovascular complications associated with isolated elevations in systolic blood pressure as well as the safety and efficacy of interventions to reduce blood pressure. These trials have shown that systolic blood pressure can be reduced by interventions such as weight loss, restriction of dietary sodium intake, and drugs. In several new trials, the long-acting dihydropyridine calcium channel blockers have been found to be safe and effective in these patients but not superior to other drugs. As in younger individuals with hypertension, drug therapy should be targeted to address comorbidity. Education of primary care physicians concerning these new findings is the next step in reducing the morbidity and mortality of this common problem in the elderly. PMID- 11062601 TI - Hypertension in patients presenting with stroke. AB - Hypertension is a major risk factor for stroke disease. There is now some international agreement on what constitutes hypertension, and at what level of blood pressure treatment is required. Large randomised controlled trials demonstrate the benefit of reducing blood pressure for the primary and secondary prevention of stroke disease. Studies have also demonstrated the benefit of particular classes of antihypertensive agents in certain patient groups. Research is beginning to elucidate the problems of hypertension in the acute phase of ischaemic stroke and the therapeutic strategies that may be helpful. Given the significant impact of stroke disease on all health services, it remains an important priority to determine the best management of hypertension in stroke. PMID- 11062602 TI - Ambulatory blood pressure monitoring. AB - Ambulatory blood pressure monitoring (ABPM) is becoming widely accepted as a clinically useful tool for assessing cardiovascular risk in hypertensive patients, although it is not generally recognized for reimbursement in the United States. There are now six major prospective studies, all of which have shown that ABPM gives a better prediction of risk than conventional clinic measurement. A corollary of these findings is that patients with white coat hypertension have been found to be at relatively low risk. The major clinical indications include patients with newly diagnosed hypertension, suspected white coat hypertension, and refractory hypertension. White coat hypertension is common during pregnancy and may lead to unnecessary cesarean sections. PMID- 11062603 TI - Hypertension and the heart. AB - Sustained increase in arterial pressure causes left ventricular hypertrophy and adversely affects all myocardial compartments: myocytes, interstitium, and coronary vasculature. Ventricular hypertrophy significantly increases the risk for cardiovascular morbidity and mortality in hypertensive disease. Impairments in coronary circulation and ventricular fibrosis, which are an essential part of hypertensive disease, contribute to that increased risk. This report discusses the mechanisms of hypertension-induced myocardial collagen accumulation and impairments in coronary hemodynamics. Particular attention is given to the interaction of hypertension and aging because aging aggravates hypertensive changes and the incidence of hypertension increases with aging. The effect of therapy on hypertension-induced ventricular fibrosis and impairment in coronary hemodynamics and the risk associated with these changes are also discussed. PMID- 11062604 TI - Acute-phase serum amyloid A production by rheumatoid arthritis synovial tissue. AB - Acute-phase serum amyloid A (A-SAA) is a major component of the acute-phase response. A sustained acute-phase response in rheumatoid arthritis (RA) is associated with increased joint damage. A-SAA mRNA expression was confirmed in all samples obtained from patients with RA, but not in normal synovium. A-SAA mRNA expression was also demonstrated in cultured RA synoviocytes. A-SAA protein was identified in the supernatants of primary synoviocyte cultures, and its expression colocalized with sites of macrophage accumulation and with some vascular endothelial cells. It is concluded that A-SAA is produced by inflamed RA synovial tissue. The known association between the acute-phase response and progressive joint damage may be the direct result of synovial A-SAA-induced effects on cartilage degradation. PMID- 11062605 TI - Active synovial matrix metalloproteinase-2 is associated with radiographic erosions in patients with early synovitis. AB - INTRODUCTION: In cancer the gelatinases [matrix metalloproteinase (MMP)-2 and MMP 9] have been shown to be associated with tissue invasion and metastatic disease. In patients with inflammatory arthritis the gelatinases are expressed in the synovial membrane, and have been implicated in synovial tissue invasion into adjacent cartilage and bone. It is hypothesized that an imbalance between the activators and inhibitors of the gelatinases results in higher levels of activity, enhanced local proteolysis, and bone erosion. OBJECTIVES: To determine whether the expression and activity levels of MMP-2 and MMP-9, and their regulators MMP-14 and tissue inhibitor of metalloproteinase (TIMP), are associated with early erosion formation in patients with synovitis of recent onset. PATIENTS AND METHOD: A subset of 66 patients was selected from a larger early synovitis cohort on the basis of tissue availability for the study of synovial tissue and serum gelatinase expression. Patients with peripheral joint synovitis of less than 1 years' duration were evaluated clinically and serologically on four visits over a period of 12 months. At the initial visit, patients underwent a synovial tissue biopsy of one swollen joint, and patients had radiographic evaluation of hands and feet initially and at 1year. Serum MMP 1, MMP-2, MMP-9, MMP-14, and TIMP-1 and TIMP-2 levels were determined, and synovial tissue was examined by immunohistology for the expression of MMP-2 and MMP-9, and their molecular regulators. Gelatinolytic activity for MMP-2 and MMP-9 was quantified using a sensitive, tissue-based gel zymography technique. Four healthy individuals underwent closed synovial biopsy and their synovial tissues were similarly analyzed. RESULTS: Of the 66 patients studied, 45 fulfilled American College of Rheumatology criteria for rheumatoid arthritis (RA), with 32 (71%) being rheumatoid factor positive. Of the 21 non-RA patients, seven had a spondylarthropathy and 14 had undifferentiated arthritis. Radiographically, 12 of the RA patients had erosions at multiple sites by 1 year, whereas none of the non RA patients had developed erosive disease of this extent. In the tissue, latent MMP-2 was widely expressed in the synovial lining layer and in areas of stromal proliferation in the sublining layer and stroma, whereas MMP-9 was expressed more sparsely and focally. MMP-14, TIMP-2, and MMP-2 were all detected in similar areas of the lining layer on consecutive histologic sections. Tissue expression of MMP-14, the activator for pro-MMP-2, was significantly higher in RA than in non-RA patients (8.4 +/- 5 versus 3.7 +/- 4 cells/high-power field; P = 0.009). In contrast, the expression of TIMP-2, an inhibitor of MMP-2, was lower in the RA than in the non-RA samples (25 +/- 12 versus 39 +/- 9 cells/high-power field; P = 0.01). Synovial tissue expressions of MMP-2, MMP-14, and TIMP-2 were virtually undetectable in normal synovial tissue samples. The synovial tissue samples of patients with erosive disease had significantly higher levels of active MMP-2 than did those of patients without erosions (Fig. 1). Tissue expression of MMP-2 and MMP-9, however, did not correlate with the serum levels of these enzymes. With the exception of serum MMP-2, which was not elevated over normal, serum levels of all of the other MMPs and TIMPs were elevated to varying degrees, and were not predictive of erosive disease. Interestingly, MMP-1 and C-reactive protein, both of which were associated with the presence of erosions, were positively correlated with each other (r = 0.42; P < 0.001). DISCUSSION: MMP-2 and MMP-9 are thought to play an important role in the evolution of joint erosions in patients with an inflammatory arthritis. Most studies have concentrated on the contribution of MMP-9 to the synovitis, because synovial fluid and serum MMP-9 levels are markedly increased in inflammatory arthropathies. Previously reported serum levels of MMP-9 have varied widely. In the present sample of patients with synovitis of recent onset, serum MMP-9 levels were elevated in only 21%. Moreover, these elevations were not specific for RA, the tissue expression of MMP-9 was focal, and the levels of MMP-9 activity were not well correlated with early erosions. Although serum MMP-2 levels were not of prognostic value, high synovial tissue levels of MMP-2 activity were significantly correlated with the presence of early erosions. This may reflect augmented activation of MMP-2 by the relatively high levels of MMP-14 and low levels of TIMP-2 seen in these tissues. We were able to localize the components of this trimolecular complex to the synovial lining layer in consecutive tissue sections, a finding that is consistent with their colocalization. In conclusion, we have provided evidence that active MMP-2 complexes are detectable in the inflamed RA synovium and may be involved in the development of early bony erosions. These results suggest that strategies to inhibit the activation of MMP 2 may have the potential for retarding or preventing early erosions in patients with inflammatory arthritis. PMID- 11062607 TI - The foundation for soaring to greater heights. PMID- 11062606 TI - Specificity of T cells in synovial fluid: high frequencies of CD8(+) T cells that are specific for certain viral epitopes. AB - INTRODUCTION: Epstein-Barr virus (EBV) is transmitted orally, replicates in the oropharynx and establishes life-long latency in human B lymphocytes. T-cell responses to latent and lytic/replicative cycle proteins are readily detectable in peripheral blood from healthy EBV-seropositive individuals. EBV has also been detected within synovial tissue, and T-cell responses to EBV lytic proteins have been reported in synovial fluid from a patient with rheumatoid arthritis (RA). This raises the question regarding whether T cells specific for certain viruses might be present at high frequencies within synovial fluid and whether such T cells might be activated or able to secrete cytokines. If so, they might play a 'bystander' role in the pathogenesis of inflammatory joint disease. OBJECTIVES: To quantify and characterize T cells that are specific for epitopes from EBV, cytomegalovirus (CMV) and influenza in peripheral blood and synovial fluid from patients with arthritis. METHODS: Peripheral blood mononuclear cells (PBMCs) and synovial fluid mononuclear cells (SFMCs) were obtained from patients with inflammatory arthritis (including those with RA, osteoarthritis, psoriatic arthritis and reactive arthritis). Samples from human leucocyte antigen (HLA)-A2 positive donors were stained with fluorescent-labelled tetramers of HLA-A2 complexed with the GLCTLVAML peptide epitope from the EBV lytic cycle protein BMLF1, the GILGFVFTL peptide epitope from the influenza A matrix protein, or the NLVPMVATV epitope from the CMV pp65 protein. Samples from HLA-B8-positive donors were stained with fluorescent-labelled tetramers of HLA-B8 complexed with the RAKFKQLL peptide epitope from the EBV lytic protein BZLF1 or the FLRGRAYGL peptide epitope from the EBV latent protein EBNA3A. All samples were costained with an antibody specific for CD8. CD4+ T cells were not analyzed. Selected samples were costained with antibodies specific for cell-surface glycoproteins, in order to determine the phenotype of the T cells within the joint and the periphery. Functional assays to detect release of IFN- or tumour necrosis factor (TNF)- were also performed on some samples. RESULTS: The first group of 15 patients included 10 patients with RA, one patient with reactive arthritis, one patient with psoriatic arthritis and three patients with osteoarthritis. Of these, 11 were HLA-A2 positive and five were HLA-B8 positive. We used HLA-peptide tetrameric complexes to analyze the frequency of EBV-specific T cells in PBMCs and SFMCs (Figs 1 and 2). Clear enrichment of CD8+ T cells specific for epitopes from the EBV lytic cycle proteins was seen within synovial fluid from almost all donors studied, including patients with psoriatic arthritis and osteoarthritis and those with RA. In donor RhA6, 9.5% of CD8+ SFMCs were specific for the HLA-A2 restricted GLCTLVAML epitope, compared with 0.5% of CD8+ PBMCs. Likewise in a donor with osteoarthritis (NR4), 15.5% of CD8+ SFMCs were specific for the HLA-B8 restricted RAKFKQLL epitope, compared with 0.4% of CD8+ PBMCs. In contrast, we did not find enrichment of T cells specific for the HLA-B8-restricted FLRGRAYGL epitope (from the latent protein EBNA3A) within SFMCs compared with PBMCs in any donors. In selected individuals we performed ELISpot assays to detect IFN- secreted by SFMCs and PBMCs after a short incubation in vitro with peptide epitopes from EBV lytic proteins. These assays confirmed enrichment of T cells specific for epitopes from EBV lytic proteins within synovial fluid and showed that subpopulations of these cells were able to secrete proinflammatory cytokines after short-term stimulation. We used a HLA-A2/GILGFVFTL tetramer to stain PBMCs and SFMCs from six HLA-A2-positive patients. The proportion of T cells specific for this influenza epitope was low (<0.2%) in all donors studied, and we did not find any enrichment within SFMCs. We had access to SFMCs only from a second group of four HLA-A2-positive patients with RA. A tetramer of HLA-A2 complexed to the NLVPMVATV epitope from the CMV pp65 protein reacted with subpopulations of CD8+ SFMCs in all four donors, with frequencies of 0.2, 0.5, 2.3 and 13.9%. SFMCs from all four donors secreted TNF after short-term incubation with COS cells transfected with HLA-A2 and pp65 complementary DNA. We analyzed the phenotype of virus-specific cells within PBMCs and SFMCs in three donors. The SFMC virus specific T cells were more highly activated than those in PBMCs, as evidenced by expression of high levels of CD69 and HLA-DR. A greater proportion of SFMCs were CD38+, CD62L low, CD45RO bright, CD45RA dim, CD57+ and CD28- when compared with PBMCs. DISCUSSION: This work shows that T cells specific for certain epitopes from viral proteins are present at very high frequencies (up to 15.5% of CD8+ T cells) within SFMCs taken from patients with inflammatory joint disease. This enrichment does not reflect a generalized enrichment for the 'memory pool' of T cells; we did not find enrichment of T cells specific for the GILGFVFTL epitope from influenza A or for the FLRGRAYGL epitope from the EBV latent protein EBNA3A, whereas we found clear enrichment of T cells specific for the GLCTLVAML epitope from the EBV lytic protein BMLF1 and for the RAKFKQLL epitope from the EBV lytic protein BZLF1. The enrichment might reflect preferential recruitment of subpopulations of virus-specific T cells, perhaps based on expression of selectins, chemokine receptors or integrins. Alternatively, T cells specific for certain viral epitopes may be stimulated to proliferate within the joint, by viral antigens themselves or by cross-reactive self-antigens. Finally, it is theoretically possible that subpopulations of T cells within the joint are preferentially protected from apoptotic cell death. Whatever the explanation, the virus-specific T cells are present at high frequency, are activated and are able to secrete proinflammatory cytokines. They could potentially interact with synoviocytes and contribute to the maintenance of inflammation within joints in many different forms of inflammatory arthritis. PMID- 11062608 TI - Congenital idiopathic clubfoot. PMID- 11062609 TI - Let them know who we are. PMID- 11062610 TI - Assessment of the geriatric orthopaedic trauma patient. AB - Elderly clients may suffer orthopaedic trauma from falls, motor vehicle accidents, abuse, or crimes. Falls are the most common reason for orthopaedic injury. While these same traumas occur in the young, older adults may not have the physiologic reserves necessary to promote healing or to prevent or recover from complications. Injuries sustained from trauma in combination with physiologic changes and comorbidity in the aged make orthopaedic trauma a significant health problem in older adults and a major treatment challenge. This article discusses diversity issues, physiologic changes, and assessment of the elderly that will help guide the orthopaedic nurse in providing care to elderly patients who experience orthopaedic trauma. PMID- 11062611 TI - Nonsteroidal antiinflammatory drugs: benefits, risks, and COX-2 selectivity. AB - Nonsteroidal antiinflammatory drugs (NSAIDs) are the most frequently prescribed class of medication for arthritis and other musculoskeletal disorders. NSAIDs block prostaglandin production, thereby reducing pain and inflammation, but may also cause significant side effects, particularly ulcers in stomach and duodenum. Some risk factors include age, previous history of ulcer, and high dose of NSAID. Synthetic prostaglandins, H2 blockers, and proton pump inhibitors have been employed to reduce risks with varying degrees of success. New NSAIDs that block only prostaglandins at sites of inflammation (COX-2 selective NSAIDs) may be significantly safer than traditional NSAIDs. PMID- 11062612 TI - Transporting children with orthopaedic conditions and surgeries. AB - While motor vehicle crashes remain one of the leading causes of death and injury to children, proper and consistent use of child safety seats and safety belts can greatly improve children's chances of survival in motor vehicle crashes. Children with orthopaedic conditions and children after orthopaedic surgery, however, may be restricted to specialized child restraint options that are capable of providing proper fit and safe travel for a period of time after the procedure. To assure proper selection and avoid secondary injury, parents and health care professionals need to be aware of best practices for transporting children postoperatively and of restraint options that exist for children with orthopaedic conditions. PMID- 11062613 TI - Evaluation of the fall prevention program in an acute care setting. AB - PURPOSE: To evaluate the effectiveness of a Falls Prevention Program (FPP) in reducing patient fall rate and to examine predictors of falls. SAMPLE: A nonrandom, stratified convenience sample of 292 subjects was selected from medical-surgical/critical care patients at a large community hospital system. The sample included 101 patients who fell prior to the FPP, 98 patients who fell after FPP, and 93 patients who did not fall after FPP. METHODS: A retrospective, preimplementation/long-term, postimplementation, comparative, descriptive design was used to address differences in patients who fell before and after FPP. Prediction factors associated with incidence of falls were assessed using a sample that included patients who fell and patients who did not fall in 1995. Data were collected about the patients and the fall incidents via a retrospective chart and incident report review. FINDINGS: No decrease in patient fall rate was found between patients who fell before and after implementation of the FPP. Patients tended to fall attempting to get out of the bed, suffering no injury. Model testing of the linear results of patients who fell and patients who did not fall after the implementation of the FPP demonstrated that fall prediction factors included age 60 or over, impaired memory, muscle weakness, and need of ambulatory assistance (Log linear chi-square: 6.048, df 4, p = 0.811 (p > .05). CONCLUSION: Identification of patients who exhibit characteristics of the fall prediction model may be useful in reducing falls in medical-surgical/critical care patients. Further testing of the four-factor model with subsequent inclusion of focused interventions may impact the incidence of falls. PMID- 11062614 TI - Traditional Chinese medicine in orthopaedic nursing. AB - Increasing popularity of traditional Chinese medicine (TCM) by the American public warrants an examination of its role in all of nursing. Many people seeking alternative treatments do so out of frustration with the inability of Western medicine to help with chronic illnesses. Thus, Western practitioners need a basic understanding of these alternative modalities to help their clients make informed choices about their health care. This article describes the underlying theory of TCM, assessments, diagnostics, and treatments found in TCM, as well as uses of TCM in orthopaedic nursing. Also included are research considerations as well as precautions in the use of TCM. PMID- 11062615 TI - Foot care: focus on the elderly. AB - Problems in the foot develop as a result of the aging process or systemic disease. Common causes of pain and disability in the elderly are nail and skin problems, predominantly corns and calluses, along with circulatory and structural problems. Because patients with orthopaedic conditions may have preexisting foot problems, it is important for nurses to distinguish between minor foot problems that can easily be treated and more serious conditions that require referral to a specialist. This article discusses pathophysiology of the aging foot, a comprehensive foot assessment, common foot problems with nursing interventions, pressure relief and shoewear, and nurses' qualifications for providing foot and nail care. PMID- 11062616 TI - Perioperative uncertainty and state anxiety of orthopaedic surgical patients. AB - PURPOSES: The purposes of this study were to determine (a) the relationship between preoperative uncertainty and anxiety of orthopaedic surgical patients in three adult developmental stages (young, middle, older) and (b) the differences in uncertainty and anxiety among the adult developmental stages, gender, and acuity levels of the disorder. DESIGN: Correlational and comparative with descriptive survey. SAMPLE: 106 adult orthopaedic surgical patients. METHODS: Subjects were contacted 1 to 2 weeks preoperatively during their preadmission visit or upon early admission the day of surgery. FINDINGS: All patients responded similarly in terms of having moderate levels of preoperative uncertainty and anxiety. There were no statistically significant differences among developmental stages, gender, or acuity levels for these orthopaedic patients. IMPLICATIONS FOR NURSING PRACTICE: Nurses need to be aware that adult orthopaedic patients such as these subjects may have moderate levels of uncertainty and anxiety prior to surgery. In addition to physical preparation for surgery, nurses need to help patients with their emotional concerns. The incorporation of preoperative psychoeducational interventions and improved communication into plans of care for all orthopaedic surgical patients is recommended. PMID- 11062617 TI - What does it mean to blow the whistle? AB - Whistle blowing causes people to pay attention. An alarm has been sounded drawing attention to a particular set of circumstances. When nurses blow the whistle because of unethical or incompetent colleagues, they are going public with this information. Yet, even though nurses are able to provide an ethical justification for the action, not all will look favorably on what has been done. This article examines the concept of whistle blowing, its relationship to advocacy, and some consequences of whistle blowing. This article provides recommendations for educators, health care agencies, and nurses as ways to support nurses who choose to blow the whistle. PMID- 11062618 TI - Introducing ... Dottie Roberts. Interview by Jessica Roberts. PMID- 11062619 TI - Alternative or complementary therapies: hocus pocus or in-between? PMID- 11062620 TI - Gene therapy in orthopaedics. AB - Genes are composed of deoxyribonucleic acid (DNA), the hereditary material of all nucleated cells. One way in which genes function is to direct the synthesis of specific proteins. When a gene is transferred to and expressed within a cell, the recipient cell produces the protein encoded by the transferred gene. This process forms the basis for gene therapy, which can be defined as the transfer of genes to patients for therapeutic purposes. Both genetic and acquired disorders may be treated, or even cured, by gene therapy. Potential orthopaedic applications include the treatment of arthritis, tumors, osteoporosis, and genetic diseases such as osteogenesis imperfecta, as well as the enhancement of tissue repair and regeneration. Impressive preclinical progress has been made in several of these areas. A phase I clinical trial of gene therapy for rheumatoid arthritis has just been completed. Orthopaedic gene therapy should become a clinical reality during the next decade. PMID- 11062621 TI - Epidural analgesia for postoperative orthopaedic pain. AB - It is well known that traditional postoperative pain management techniques often fail to relieve pain in nearly 50% of postoperative patients. The etiology of poor pain management is rooted in the fears of addiction, overmedication, and the time associated with obtaining controlled substances and administering them to patients. Patients being treated with neuroaxial analgesia (epidural and intrathecal drug administration) are appearing more frequently on orthopaedic nursing units. It is therefore imperative that nurses are familiar with the care of patients with epidural catheters. This article discusses the relevant anatomy, physiology, pharmacology, contraindications, and complications associated with neuroaxial analgesia, with special emphasis placed on the orthopaedic patient. PMID- 11062622 TI - Introduction to the nursing research process. AB - Many nurses have expressed an interest in conducting a nursing research project, but some have not had sufficient or recent exposure to the process of how to develop a research study. The nursing research process can be outlined using the nursing process steps of assessment, planning, intervention and evaluation. In the assessment phase the problem is identified, the literature is reviewed, and the variables are identified. In the planning phase the research question or hypothesis is formulated, and decisions are made on how the variables will be measured and how the sample will be chosen. In the intervention phase the data collection occurs; in the evaluation phase the data is analyzed and interpreted and the findings are communicated. By giving nurses a detailed yet understandable plan on how to conduct nursing research, their curiosity is encouraged and the body of knowledge will grow. PMID- 11062623 TI - Intrathecal baclofen therapy and the child with cerebral palsy. AB - Children with cerebral palsy have tight, spastic muscles that often interfere with function, care, and, ultimately, quality of life. Until recently, treatment usually focused on the effects of the spasticity rather than the spasticity itself. Intrathecal Baclofen Therapy (ITB) administers a muscle-relaxing agent directly into the intrathecal space; thereby decreasing the amount of spasticity the child exhibits. This article discusses cerebral palsy and a new treatment for spasticity, ITB, and the development of an ITB program. PMID- 11062624 TI - Patients' and their spouses' needs after total joint arthroplasty: a pilot study. AB - PURPOSE: To describe the experiences and needs of patients and their spouses during hospitalization and recovery from either total hip or total knee arthroplasty. DESIGN: Descriptive. SAMPLE: A purposive sample of 5 patients and their spouses in one health sciences center. METHODS: Qualitative study using a videotaped focus group interview. FINDINGS: Content analysis revealed two perspectives of one theme: patients and their spouses need "help making transitions." Situational and role transitions that were problematic for patients reflected distress over not being able to resume activities they enjoyed within an expected time frame. Incongruence between expectations and reality was the source of distress. As a consequence of role reversal, spouses experienced feelings of insecurity and being overwhelmed. Health and illness transitions that patients experienced were also related to incongruence between expectations of the recovery period and the reality that recovery is a slow process. Pain experienced during postdischarge recovery and rehabilitation, and problems encountered when applying information and skills learned in the hospital to the home setting were sources of concern. CONCLUSION: The needs and experiences of patients and spouses after total joint arthroplasty reflect transitional change- changes in roles, relationships, abilities, and behaviors. Health care professionals can facilitate transitions by providing education that reflects "best case-worst case scenarios" so that expectations of the recovery process are realistic. By being the link between hospital and home, health care professionals can lend support to patients as they continue the recovery process. As a program evaluation strategy, focus groups provide useful information to health care professionals who are interested in the needs and expectations of health care consumers. IMPLICATIONS FOR NURSING RESEARCH: Further exploration of the needs of patients and their spouses following joint replacement surgery is warranted. Use of focus group methodology might provide additional insight into the needs of this population and suggest ways in which health care professionals can modify existing programs to help these patients and their spouses make the transitions. PMID- 11062625 TI - Qualitative research and focus group methodology. PMID- 11062626 TI - Shingles update: common questions in caring for a patient with shingles. AB - Varicella zoster virus (VZV) is a herpes virus that can cause two distinct clinical diseases, chickenpox and shingles. Primary infection of varicella, often called chickenpox, results in a generalized eruption of a vesicular exanthematous rash which is usually seen in children and is highly contagious. This virus (VZV) can then become latent and later reactivate causing herpes zoster, commonly known as shingles. Shingles is usually a localized phenomenon often seen in adults and is usually less contagious. The following is a discussion of infection control questions most commonly asked regarding the care of a patient with shingles. PMID- 11062627 TI - Vitamin D deficiency rickets in children: prevalence and need for community education. AB - Nutritional rickets caused by a deficiency in vitamin D can be a crippling disease and is still prevalent today. Many feel that the vitamin D supplementation present in dairy products prevents any risk of developing vitamin D deficiency rickets. However, a combination of factors such as pigmented skin, inadequate sunlight, breastfeeding past 6 months with no supplementation, and insufficient calcium intake by the nursing mother and her baby often contribute to rickets diagnosed in children. Educating the medical community and those families at risk is critical to eradicate this preventable disease. Steps can be taken using stages of the Organizational Change Theory to involve school nurses, community health centers, and pediatricians in identifying these high-risk children. With prompt diagnosis and treatment, all symptoms can abate, with healthy bone growth resulting. PMID- 11062628 TI - The relationship between multidisciplinary discharge outcomes and functional status after total hip replacement. AB - PURPOSE: To determine changes in functional status after primary total hip replacement and to explore the relationship between functional status and pain and mobility at time of hospital discharge. DESIGN: Descriptive correlational with a prospective cohort. SAMPLE: 21 elective primary total hip arthroplasty patients, ages 40 to 78, in an academic health center. METHOD: Subjects' functional status was measured using the Sickness Impact Profile and the hip outcome tool (Revised Hip Type Specification Tool 13.1), completed by face-to face interview approximately 2 weeks prior to hospitalization. Pain and mobility were assessed at time of discharge, and the functional status measures were repeated at 3 months after hospital discharge by mail. FINDINGS: Statistically significant changes were found for physical and psychosocial dimensions of functional status, using the Sickness Impact Profile and the hip outcome tool, between the preoperative and postoperative measures. A significant relationship between mobility and pain at the time of discharge and functional status was not established. IMPLICATIONS: Hospital discharge criteria related to pain and mobility should be used with caution. In addition to pain and mobility information, patient education should reflect knowledge of what can be expected by 3 months postsurgery, including expected and potential improvements in mobility, pain, and ambulation and in nonphysical dimensions, such as sleep, home management, and social interaction. Case management and discharge planning should reflect the wide variance in pain and mobility across patients at time of discharge and the expected gains in specific aspects of functional status after discharge. Research attention should be increased on the identification and application of specific discharge criteria such as pain and mobility levels, and the further development and refinement of nursing-sensitive and practical functional status outcome measures. PMID- 11062629 TI - Common herbal remedies. AB - Herbal remedies are becoming increasingly popular as people seek more effective, natural, or safer methods for treating a variety of complaints. As a result, nurses in every setting may expect to see increased numbers of patients who are using herbal products. When patients assume that the nurses will be critical of their use of herbals, they may withhold such information to avoid unpleasantness. This could place patients at risk for adverse effects, drug interactions, and complications related to ineffective treatment. Nurses who are knowledgeable about herbal products and who are open to discussion about these products can provide information and advice about safe use. The discussion in this article addresses actions, possible benefits, and dangers of the most common herbal products. Guidelines for assessing and teaching clients about herbal use are included. PMID- 11062630 TI - Introducing Marj Kulesa. PMID- 11062631 TI - The dollars and sense of professional development. PMID- 11062632 TI - How drug-resistant microorganisms affect nursing. AB - Although antibiotics have improved the mortality and morbidity associated with infectious disease, antibiotic mismanagement has created drug-resistant microorganisms, which are present in the environment outside the hospital. In the hospital, drug-resistant microorganisms require Contact Isolation, a transmission based isolation recommended by the Centers for Disease Control and Prevention. The nurse's role in preventing the spread of drug-resistant microorganisms includes using appropriate infection control and isolation guidelines (especially handwashing), working with other health care workers to maintain appropriate infection control standards at all times, and teaching patients and families about infection control and antibiotic management. PMID- 11062633 TI - Management of lower extremity riding lawn mower injuries in children. AB - Eight children are injured by riding lawn mowers every day. The child, usually a bystander or passenger on the mower, can sustain life-threatening and limb threatening injuries. Multidisciplinary care must be available to manage the numerous issues presented by the unique circumstance of a child with a severe injury in the acute and chronic settings. Whether the limb is salvaged or amputated, the ultimate goal is optimal functional outcome for the patient. We have developed a team approach to address these injuries from their onset until patient maturity, maximizing our ability to achieve this goal. PMID- 11062634 TI - Prevention and management of postoperative nausea and vomiting. AB - PURPOSE: To identify factors associated with the occurrence of postoperative nausea and vomiting and to identify the effectiveness of independent and interdependent nursing interventions for the prevention and management of postoperative nausea and vomiting. DESIGN: Descriptive. SAMPLE: A convenience sample of 300 patients, 18 years or older, who had surgery with general anesthesia (excluding persons having ear or gastrointestinal surgery), at either campus of a large metropolitan medical center. METHOD: A combination of closed chart review and data collection by staff nurses assigned to the patients in the postoperative period using a standardized data collection tool. FINDINGS: Data was analyzed using a combination of descriptive and parametric statistics. There was a 39% incidence of nausea and vomiting overall. Women were two times more likely to experience nausea and twice as likely to experience severe nausea (statistical significance of p = .0001). In addition, persons with surgery lasting 2 hours or greater were twice as likely to experience nausea (statistical significance of p = .002). Independent nursing interventions, such as placing a cool washcloth on a patient's forehead, were used more often than medication as an initial strategy. The most frequently used interventions were increasing the i.v. fluids and having the patient take deep breaths. There was no pattern to the nursing assessments. CONCLUSION: Women and those patients with surgery lasting greater than 2 hours should be assessed for nausea in the postoperative period more frequently. Assessments need to be done during the first 2 hours after surgery, at 7-8 hours after surgery, and when there is an increase in activity, such as the first time out of bed. Nurses do not routinely document assessments for postoperative nausea. In addition, some misconceptions exist. Formal and informal education for nurses, physicians, and patients on the incidence, predisposing factors, and treatment options for postoperative nausea needs to occur. This should include reflection on the misconceptions, the need for ongoing patient assessment, and a review of drug and nondrug therapies. IMPLICATIONS FOR NURSING RESEARCH: Further research is needed to better describe the present practice patterns in the treatment of postoperative nausea and vomiting, including a comparison of inpatient and outpatient settings. The relative effectiveness of specific nursing interventions should be measured, such as i.v. fluids, deep breathing, and antiemetic use. In addition, the evaluation of the effectiveness of the use of a research protocol would be useful. PMID- 11062635 TI - The pilot study. PMID- 11062636 TI - Care of the patient with a tracheostomy. AB - Patients with tracheostomies may not be commonly admitted to orthopaedic units. Since this is an infrequently seen therapy in this population, and because the potential complications carry very high risks, orthopaedic nurses may be unfamiliar or uncomfortable with the nursing care needs of these patients. This article reviews the indications for tracheostomy tube insertion and the resulting physiologic changes that occur. Nursing assessment and tracheostomy care is described along with detailed information on potential complications and the associated nursing interventions. PMID- 11062637 TI - Tips for writing a NAON research grant proposal. AB - This article is intended to help the novice nurse researcher prepare a research grant proposal for the National Association of Orthopaedic Nurses (NAON). It summarizes the grant guidelines and describes each component of the research proposal. The components include the abstract, statement of the problem, literature review, purpose, design, methodology, timetable and budget. NAON's research priorities are included as well as contact information for the organization and tips for survival. PMID- 11062638 TI - Osteoporosis: a disease management opportunity. AB - Advances in the ability to detect and effectively treat osteoporosis lead to questions about when testing and treatment should be initiated. Disease management provides answers with the promise of both cost-effective use of resources and improved health outcomes. Applying the characteristics and principles of disease management to osteoporosis provides a powerful rationale for a population-based approach to this disease. Disease management makes it possible to systematically identify persons at risk, intervene with prevention and treatment programs, and measure clinical, quality of life, and economic outcomes. Clinical guidelines are a critical element in disease management. This article presents clinical guidelines developed recently by national medical and public health experts based on a review of available research and clinical evidence. PMID- 11062639 TI - Relationships among self-efficacy, outcome expectancy, and postoperative behaviors in total joint replacement patients. AB - PURPOSE: To determine whether self-efficacy and outcome expectancy predicted performance of postoperative leg exercises and ambulation among persons recovering from total hip or total knee replacement. DESIGN: Descriptive correlational. SAMPLE: 50 subjects recovering from their first unilateral total hip or total knee replacement at a large midwestern hospital who had completed a preoperative education course. METHODS: On postoperative day 1, subjects who correctly performed leg exercises and could properly ambulate with a walker were administered the Self-Efficacy Scale and the Outcome Expectancy Scale. On postoperative day 2, frequency and repetitions of leg exercises were measured by self-report. Ambulation was measured by distance walked in feet. FINDINGS: Self efficacy was the sole predictor of the dependent variables, accounting for 8% to 33% of the variance. Higher levels of self-efficacy were correlated with longer distances in ambulation and with higher frequency and more repetitions of leg exercises. IMPLICATIONS FOR NURSING PRACTICE: Solely providing a preoperative education course is not sufficient to ensure performance of postoperative behaviors in total joint replacement patients. Patients' self-efficacy beliefs may need to be considered when planning a preoperative educational program for patients undergoing total joint replacement surgery. This study's situation specific measure of self-efficacy could be used in the clinical setting to identify patients with low levels of self-efficacy who may be less likely to perform preventive postoperative behaviors. PMID- 11062640 TI - Enhancing documentation for the orthopaedic patient. AB - A need for integrated, interdisciplinary documentation led to an assessment and evaluation of our current documentation system. This article describes the experiences of a two-hospital health care system in piloting and implementing documentation changes on an orthopaedic unit. Outcomes, conclusions, and a copy of the total joint plan of care are also included. PMID- 11062642 TI - Introducing Cynthia Gonzalez. PMID- 11062641 TI - Clinical research: what do patients understand? AB - An increasing amount of clinical research is being conducted to test the efficacy and effectiveness of new medications, therapies, and medical procedures. These studies often require that patients be invited to be subjects. However, what do patients really understand whenever they are asked to participate in clinical research? Are patients aware that research is clearly different from treatment? Are they aware that their participation in research is to benefit future patients? Despite the desire of clinical agencies to support research, there is the concern about how they can best protect the rights of patients who are subjects in this research. The author briefly discusses clinical research, patients as vulnerable subjects, the therapeutic misconception, and informed consent. In addition, recommendations are provided to help assure that the rights of patients who are research subjects are protected. PMID- 11062643 TI - 2000 Ostomy Wound Management Buyers Guide. PMID- 11062644 TI - [Detection of cancer or prevention of metastases?]. PMID- 11062645 TI - [Tobacco and cancer: the role of the nurse]. PMID- 11062646 TI - [The nurse's role in follow-up and chemotherapy at the patient's home]. PMID- 11062647 TI - [City-hospital: chemotherapy at home]. PMID- 11062648 TI - [Care of patients operated on for breast cancer with immediate or delayed reconstructive surgery]. PMID- 11062649 TI - [Caring for patients who underwent radical surgery for gynecologic cancer]. PMID- 11062650 TI - [How to improve organization of blocks]. PMID- 11062651 TI - [Industrial medicine in the management of epilepsy]. PMID- 11062652 TI - [Venous insufficiency, heavy legs, embolism]. PMID- 11062653 TI - [National protocol on the organization of emergency services in schools and local public educational establishments]. PMID- 11062654 TI - [Alzheimer disease: France adjusts the balance]. PMID- 11062655 TI - [Therapeutic education: a "plus" in the management of the diabetic patient]. PMID- 11062656 TI - [Diabetic neuropathy: danger to the foot]. PMID- 11062657 TI - [A chance in the pursuit of success]. PMID- 11062658 TI - [Materials, equipment: technologic progress (diabetes)]. PMID- 11062659 TI - [Psychologic support for diabetic patients]. PMID- 11062660 TI - [The nurse at a vacation camp for diabetic children]. PMID- 11062661 TI - Killing nursing. PMID- 11062662 TI - Is the personal computer dead? PMID- 11062663 TI - I.v. push at home? AB - Patient administration of i.v. push antibiotics at home is one of the latest moves in this climate of shortened hospital stays and cost-cutting. The nurse's assessment, instruction, and monitoring can make the difference between a smooth course of therapy and one marked by avoidable complications. PMID- 11062664 TI - Letterman knows why I chose nursing. PMID- 11062665 TI - Oncology today: ovarian cancer. AB - While researchers work to improve treatment and come up with an accurate way to detect the disease early on, ovarian cancer will kill 14,000 women in the United States this year. Here's an update on this particularly insidious form of cancer: how it's managed today and treatment options for the future. PMID- 11062666 TI - ED nursing--a whole new world. AB - If life isn't as exciting and fast-paced as you'd like, consider a move to the emergency department, where the challenges are many and the rewards immense. If you've toyed with the idea but are intimidated by all you'll need to know, here are a few hints for a smooth transition. PMID- 11062667 TI - Exploring chest drain options. AB - The variety of chest drainage systems now available makes it more important than ever for nurses to understand what's out there and how these systems work. Nurses are responsible for managing chest drains and need to know which drain is best suited for which situation. PMID- 11062668 TI - Chest drainage systems. PMID- 11062669 TI - Nurses and suicide: the risk is real. PMID- 11062670 TI - Treatment options for acute migraine. PMID- 11062671 TI - Exploring the world of nursing on the Net. PMID- 11062672 TI - [Objectives, indications and modalities of detoxification of the alcoholic patient]. PMID- 11062673 TI - [The process of accreditation is clearly launched]. PMID- 11062674 TI - [Hospitalization of young persons committing suicide in France]. PMID- 11062675 TI - [Bosnia, experience of war, a step into the future]. PMID- 11062676 TI - [Prevention of lumbago in hospital environment]. PMID- 11062677 TI - [To consider lumbago as a professional disorder]. PMID- 11062678 TI - [Taking care of the alcoholic patient]. PMID- 11062679 TI - [Alcoholism in all its stages]. PMID- 11062680 TI - [Suffering of the caretaker staff treating addicts]. PMID- 11062681 TI - [Therapeutic relationship with the alcoholic patient]. PMID- 11062682 TI - [Taking over care of alcoholics by the nurse: what is the procedure?]. PMID- 11062683 TI - [It is not the mother who takes care of the man...]. PMID- 11062684 TI - ["Ecologic" approach to the alcoholic patient]. PMID- 11062685 TI - [The school nurse: her role in the prevention of alcohol dependence]. PMID- 11062686 TI - Oral presentations in science and medicine. An art in decline? AB - The presentation of understandable and stimulating lectures on difficult scientific and medical issues is in our opinion an art in decline. This disconcerting trend has, however, received little attention. In this essay, we expose the most common fallacies and pitfalls encountered in poor lectures, discuss the underlying reasons why they occur and consider some remedies to improve the situation. The status of oral communication should in our opinion be upgraded. To give a first rate, lucid lecture is an important service to the scientific community and should be recognised as such. When principal speakers at symposia are selected, and when researchers are considered for academic positions, documented proficiency in oral communication should be given more weight than is currently the case. This essay does not purport to be a treatise on rhetoric. We believe that a more efficient approach is to focus attention on the common blunders. This we have done in a provocative manner to invite comments and discussion. PMID- 11062687 TI - Identification of THW, a putative new tumor suppressor gene. AB - Differential Display between metastasizing and non-metastasizing human melanoma cell lines NMCL-1 and 530 led to the identification of a putative transmembrane receptor, designated as THW, which is down-regulated in the metastasizing cell line. THW is composed of 193 aa, exhibits four putative transmembrane domains and does not reveal any homology to other proteins. Down-regulation of THW is correlated with metastatic capacity of melanoma cell lines. THW was down regulated in mammary carcinoma cell lines compared with cell lines derived from non-malignant mammary epithelium, in pancreas cell lines derived from metastases compared with a cell line derived from a primary tumor and in several tumor tissues compared with normal tissues. The function of THW as a tumor suppressor is emphasized by the location of the THW gene on chromosome 6q. LOH for this region has been reported in malignant melanoma and prostate, pancreas, uterine and mammary carcinomas. PMID- 11062688 TI - Adenovirus-mediated suicide gene therapy for bladder cancer: comparison of the cytomegalovirus- and Rous sarcoma virus-promoter. AB - BACKGROUND: To compare efficacy and toxicity of the human cytomegalovirus immediate-early (CMV) promoter and the Rous-sarcoma-virus (RSV) promoter to express thymidine kinase (tk) for adenovirus-mediated suicide gene therapy of experimental bladder cancer in vivo and in vitro. MATERIALS AND METHODS: In vitro: 3 human (5637, RT-4 and TCC-SUP) and one murine (MBT-2) bladder cancer cell line were exposed to ADV/RSV-tk or ADV/CMV-tk vectors and cell survival was determined. In vivo: Subcutaneous tumors were established and adenovirus vectors were injected 10 days later. RESULTS: In vitro: ADV/CMV-tk was up to 4 times more potent in terms of cell killing than ADV/RSV-tk. In vivo: ADV/CMV-tk had a three fold higher antitumor potency per viral particle as compared to ADV/RSV-tk. Higher doses of ADV/CMV-tk caused treatment-associated hepatotoxicity. CONCLUSIONS: Our results confirm the efficacy of adenovirus-mediated tk suicide gene therapy in the treatment of experimental bladder cancer. Dose-related toxicity was greater with the use of ADV/CMV-tk, but lower doses achieved the same efficacy as ADV/RSV-tk. PMID- 11062689 TI - Controlled-release of doxorubicin from poly(lactide-co-glycolide) microspheres significantly enhances cytotoxicity against cultured AIDS-related Kaposi's sarcoma cells. AB - Subsequent to the introduction of highly active antiretroviral therapy (HAART), there has been a reduction in HIV viral titers and a concomitant decrease in AIDS related Kaposi's sarcoma. However, as failure rates of HAART approach 30%, concerns arise regarding resurgence in AIDS-KS. Current AIDS-KS therapies fail to provide sustained remissions and yet also result in significant morbidity. Although partially effective, systemic chemotherapy is particularly debilitating to AIDS patients. In this report, we examined the co-incubation of AIDS-KS cells with doxorubicin which was slowly delivered from biodegradable, locally injectable, controlled-release poly(lactide-co-glycolide) (PLGA) microspheres. Local drug delivery systems such as PLGA microspheres can sustain therapeutic intralesional concentrations while minimizing deleterious systemic side effects, providing a pharmacologic advantage at the treatment site. Our data show that controlled release from PLGA microspheres augments doxorubicin cytotoxicity towards AIDS-KS cells without increasing toxicity in nonlesional cells from the AIDS-KS donors. Electron microscopic analysis revealed that PLGA microspheres possess a strong affinity for cell membranes, facilitating doxorubicin delivery to redox-sensitive cell membrane sites. Consistent with their speculated endothelial cell lineage, some of the AIDS-KS cells appeared to engulf microspheres via phagocytosis. Our results suggest that PLGA controlled-release doxorubicin microspheres have potential clinical applicability in management of AIDS-KS. PMID- 11062690 TI - Inhibitory effect of erythromycin on P-glycoprotein-mediated biliary excretion of doxorubicin in rats. AB - The macrolide antibiotic erythromycin has recently been shown to overcome the resistance to anticancer drugs that results from overexpression of P glycoprotein. The present study, using erythromycin lactobionic acid as a model drug, investigated the inhibitory effects of erythromycin on the efflux of doxorubicin from P388/ADR cells expressing P-glycoprotein and on the biliary excretion mechanism of doxorubicin in rats, which is primarily mediated by P glycoprotein. Erythromycin lactobionic acid was found to inhibit the efflux of doxorubicin (5 microM) from P388/ADR cells in a concentration-dependent manner. In rats receiving constant-rate infusion of doxorubicin (30 micrograms/min), both the biliary and renal clearance of this drug dramatically decreased and its plasma concentrations increased after an intravenous injection of erythromycin lactobionic acid (100 mg/kg as erythromycin). These results suggest that erythromycin competitively inhibits P-glycoprotein-mediated biliary and renal excretion of doxorubicin. The effect of erythromycin on the biliary secretion of doxorubicin was also analyzed quantitatively by the competitive inhibition model. The computer-estimated values of Vmax/Km, Km and Ki were 8.79 ml/minute, 0.82 microgram/ml and 0.41 microgram/ml, respectively. The findings of these experiments suggest that the inhibitory effect of erythromycin on the P glycoprotein-mediated biliary excretion of doxorubicin is competitive and that combination chemotherapy of doxorubicin with erythromycin may induce toxicity as a result of increased plasma concentrations of doxorubicin. PMID- 11062691 TI - Drug-induced inhibition of the peripheral-type benzodiazepine receptor expression and cell proliferation in human breast cancer cells. AB - The peripheral-type benzodiazepine receptor (PBR) expression and localization correlate with human breast cancer cell proliferation and aggressive phenotype expression. The standardized extract of Ginkgo biloba leaves (EGb 761) and isolated ginkgolide B (GKB) were shown to decrease PBR mRNA expression in adrenal cells. We examined the effect of EGb 761 and GKB on PBR expression and cell proliferation in human breast cancer cells. EGb 761 and GKB decreased in a time- and dose-dependent manner PBR expression and cell proliferation in the highly aggressive, rich in PBR, human breast cancer cell line MDA-231 whereas they did not affect the proliferation of the non-aggressive human breast cancer cell line MCF-7, which contains very low PBR levels. This effect was reversible and not due to the antioxidant properties of the compounds tested. Using a human cDNA expression array we determined that EGb 761 treatment altered, in addition to PBR, the expression of 36 gene products involved in various pathways regulating cell proliferation. These in vitro data were further validated in an in vivo model where EGb 761 and GKB significantly inhibited the nuclear PBR expression and growth of MDA-231 cell xenografts in nude mice. Taken together, these data suggest that the manipulation of PBR expression could be used to control tumor growth and that EGb 761 and GKB, under the conditions used, exert cytostatic properties. PMID- 11062692 TI - Loss of heterozygosity in gastric neuroendocrine tumor. AB - The MEN1 gene locus is known to be partly responsible for the tumorigenesis of sporadic gastric neuroendocrine tumors, but the genetic events that drive the neoplastic process of this tumor remain largely unknown. In order to screen the tumor suppressor genes associated with the tumorigenesis of gastric neuroendocrine tumors, 15 neuroendocrine carcinomas and three carcinoid tumors in the stomach were analyzed for loss of heterozygosity (LOH) using 22 microsatellite markers. In our study, the gastric neuroendocrine tumors showed a high rate of LOH in chromosomes 8p (82%), 15q (58%), 17p (57%), llp (50%), 12p (50%) and 13q (50%). The mean fractional allelic loss (FAL) was higher in the neuroendocrine carcinoma components than in the adenocarcinoma components (0.42 versus 0.33, respectively). In four cases, the adenocarcinoma components showed discordant LOH patterns from those of the neuroendocrine counterparts in half of the informative chromosomes analyzed. Comparably, the gastric neuroendocrine carcinomas exhibited a higher LOH frequency on 8p and a lower LOH on 7q than did the gastric adenocarcinomas. It is suggested that chromosome 8p is the possible location of the tumor suppressor genes associated with the tumorigenesis of gastric neuroendocrine tumors. PMID- 11062693 TI - Apoptosis and expression of Bax and Bcl-2 in snuff- and non-snuff associated oral squamous cell carcinomas. AB - BACKGROUND: Oral squamous cell carcinoma (OSCC) is etiologically linked to tobacco and alcohol consumption. A higher frequency of p53 gene mutations was reported in snuff (toombak)-associated OSCC from the Sudan versus those from non users (Ibrahim et al., 1999, 10). MATERIALS AND METHODS: OSCC from Sudanese toombak users (n = 13) and non-users from the Sudan (n = 6) and Norway (n = 24) were analysed for bax, bcl-2 and Ki-67 immunohistochemically. Apoptosis was evaluated by the TUNEL method. The OSCC from the Sudan had previously been studied for p53 gene mutations. RESULTS: We found a higher apoptotic rate and a higher bax expression in OSCC from Norway compared with those from the Sudan (p < 0.05) irrespective of toombak use. No significant differences were detected in apoptosis, bax, bcl-2 and Ki-67 in OSCC from the Sudan in relation to toombak use or p53 gene status. CONCLUSION: In OSCC, apoptosis was associated with bax expression and was unaffected by p53 gene status or toombak use in OSCC from the Sudan. PMID- 11062694 TI - Antioxidant and free radical scavenging effects of baicalein, baicalin and wogonin. AB - Xanthine oxidase inhibitors are known to be therapeutically useful for the treatment of hepatitis and brain tumor. Baicalein, baicalin and wogonin, isolated from Scutellaria rivularis, have been reported to exhibit a strong activity on xanthine oxidase inhibition. In this study, their antioxidant activity was evaluated by modified xanthine oxidase inhibition and cytochrome c reduced methods. The results showed that the order of activity on xanthine oxidase inhibition was baicalein > wogonin > baicalin, IC50 = 3.12, 157.38 and 215.19 microM, respectively, whereas the activity on cytochrome c reduction was baicalin > wogonin > baicalein (IC50 = 224.12, 300.10 and 370.33 microM, respectively). In another study, an electron spin resonance (ESR) technique was used to further confirm the direct free radical scavenging activity. Both baicalein and baicalin demonstrated a strong activity on eliminating the superoxide radical (.O2-) (baicalein: 7.31 x 10(4) u/g; baicalin: 1.19 x 10(5) u/g). The IC50 of baicalein was 2.8 fold higher than that of baicalin. However they had no significant effect on scavenging hydroxyl radical (.OH). The present results demonstrated that baicalein and baicalin posed a different pathological pathway. The antioxidant function of baicalin was mainly based on scavenging superoxide radical whilst baicalein was a good xanthine oxidase inhibitor. PMID- 11062695 TI - Meloxicam inhibits the growth of non-small cell lung cancer. AB - Cyclooxygenase (COX)-2 has been reported to play an important role in carcinogenesis. Meloxicam (preferential COX-2 inhibitor) inhibits the growth of COX-2 positive and COX-1 negative colorectal cancer cells. We evaluated the effects of meloxicam on the growth of lung cancer cells. By reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot analysis, COX-2 but not COX-1 was expressed in human non-small cell lung cancer (NSCLC) cell lines (A549 and PC14). In human small cell lung cancer (SCLC) cell line (H841), both COX-1 and COX-2 were not detected. MTT assay and prostaglandin (PG) E2 enzyme immunoassay showed that meloxicam inhibited the growth and PGE2 production of both A549 and PC14, but not H841 cells. These findings suggest that COX-2 may play an important role in the pathogenesis and progression of NSCLC, and that meloxicam may be a useful therapeutic agents in the treatment of NSCLC. PMID- 11062696 TI - Low molecular thymic peptides stimulate human blood dendritic cells. AB - Dendritic cells are considered to be the most potent antigen-presenting cells and are thus promising new tools for the immunotherapy of cancer. They respond to various stimuli by differentiation (expression of CD83) and up-regulation of costimulatory surface molecules. Thymic peptides have immunostimulatory and immunomodulating properties. Their therapeutic potential in immunotherapy of cancer has been discussed. To test whether thymic peptides act on dendritic cells, we examined the effects of a standardized thymic peptide preparation on cultured human monocyte-derived dendritic cells. Addition of thymic peptides resulted in enhanced expression of the specific differentiation marker CD83 in a dose dependent manner. Moreover, thymic peptides induced the up-regulation of costimulatory molecules including CD86, CD80, HLA-DR and HLA-ABC. After priming with thymic peptides dendritic cells showed an enhanced expression of IL-8 and TNF-alpha mRNA and protein release. Dendritic cells stimulated with thymic peptides were able to induce proliferation of autologous T cells as measured by 3H-thymidine incorporation in mixed Lymphocyte reaction. In combination with a low dosage of keyhole limpet hemocyanin, thymic peptides showed additive effects in the up-regulation of CD83 and costimulatory surface markers. Our findings indicate that thymic peptides per se act on professional antigen-presenting cells in a stimulatory manner and were presented by these cells. Furthermore, thymic peptides enhance the response of dendritic cells to low dosages of a standard nominal antigen. Therefore, thymic peptides could improve the immunological activity especially against low amounts of endogenous antigens. PMID- 11062697 TI - Telomerase activity, Myc and Bcl-2: possible indicators of effective therapy of prostate cancer with 9-nitrocamptothecin. AB - In this report, we demonstrate the down-regulation of telomerase activity and c Myc and Bcl-2 expression during 9-nitrocamptothecin (9NC)-induced regression of human DU145 prostate tumors grown as xenografts in immunodeficient mice. These changes were not observed in tumors generated by DU145-derived cells resistant to 9NC. We suggest that telomerase activity, c-Myc and Bcl-2 can collectively serve as molecular diagnostic indicators of the effectiveness of 9NC during treatment of human prostate tumors. PMID- 11062698 TI - The human multidrug resistance gene (MDR-1): immunocytochemical detection of its expression in oral SCC. AB - A large number of oral cancer patients show poor or partial response to chemotherapy and the mechanisms are poorly understood. At present, an MDR-1 product, the P-170 glycoprotein, is the best known of the P-170 family and is involved in resistance to natural product-based chemotherapeutics, including taxanes, anthracyclines, vinca alkaloids, podophyllotoxins and camptothecins. Although several reports suggest that P-170 is clinically relevant in haematological malignancies, its role in solid tumours is not well understood. Its overexpression has been found to be correlated with the poor outcome observed in patients treated with chemotherapy and presenting drug resistance. The aim of this study was to detect the protein expression patterns of MDR-1 product by immunohistochemistry in formalin-fixed-paraffin-embedded tissues. For these reasons, 30 oral SCC and 6 healthy oral mucosa specimens were tested with anti-P 170 antibodies using standard streptavidin-biotin-peroxide technique. Immunohistochemistry demonstrated that 4 cases (66.6%) of normal oral mucosa and 24 cases (80%) of oral SCC showed positivity. Four cases (13.4%) showed strong positivity in tumour areas and complete negativity in normal epithelial cells adjacent to the tumour. No staining was observed in stromal structures, with the exception of the lymphocytic compartment that showed a strong staining as reported in literature for CD56+ and CD8+ cells. Four G1 tumours (33%) and 2 G3 tumour (33%) showed strong positivity in areas with a higher degree of differentiation. P-170 positivity in normal epithelial cells of smoker patients, in differentiated area of neoplasia and negativity or zonal positivity in undifferentiated area of tumour suggested that activation of the MDR-1 gene or selection of intrinsically multidrug resistance neoplastic cells may occur at early stages of tumorigenesis of oral cancers, before the real evidence of cellular transformation. Thus the contact with possible chemical carcinogens, such as those of tobacco smoke, may induce activation of MDR-1 gene. This study was conducted only on untreated carcinomas so for this reason it cannot indicate the real incidence of acquired multidrug resistance. The data of MDR-1 product expression by immunohistochemistry in oral SCC might suggest that an overexpression of this protein could constitute a hallmark of potential more aggressive phenotype for this type of neoplasia and a rapid method for pre screening tumours for a constitutive multidrug resistance in order to orientate the cancer treatment. PMID- 11062699 TI - Inhibitory effects of active substances isolated from Cassia garrettiana heartwood on tumor growth and lung metastasis in Lewis lung carcinoma-bearing mice (Part 1). AB - A methanol extract (500 mg/kg x 2/day) of the heartwood of Cassia garrettiana inhibited the tumor growth and metastasis to the lung in Lewis lung carcinoma (LLC)-bearing mice. We isolated the two active substances from the methanol extract of C. garrettiana: compound 1 was identified as Cassigarol A and the determination of the structure of compound 2 is now in progress. We examined the effect of the active substance (compound 1, Cassigarol A) on tumor growth and lung metastasis in LLC-bearing and primary tumor-removed mice and found that Cassigarol A (50 mg and 100 mg/kg x 2/day) inhibited the tumor growth and metastasis to the lung. In addition, Cassigarol A inhibited the plasmin activity and the formation of capillary-like networks of human umbilical vein endothelial cells (HUVECs) at concentrations of 10 to 100 microM. Therefore, it is suggested that the antitumor and antimetastatic activities of Cassigarol A might be due to the inhibition of plasmin activity and formation of tubes (angiogenesis) from HUVECs. PMID- 11062700 TI - Ferric nitrilotriacetate induced DNA and protein damage: inhibitory effect of a fermented papaya preparation. AB - The carcinogen Fe-NTA catalyzes the hydrogen peroxide-derived production of free radicals and possibly acts through a mechanism involving oxidative stress. Fermented papaya preparation (FPP) has been reported as a natural antioxidant able to prevent lipid peroxidation in vitro and in vivo. However, little is known about the antioxidant properties of FPP regarding iron-mediated oxidative damage to DNA and proteins. In the present study FPP protected supercoiled plasmid DNA against Fe-NTA plus H2O2 induced single and double strand breaks. Similar protective effects of FPP were evident when human T-lymphocytes were challenged with Fe-NTA/H2O2 and DNA damage was determined using the Comet assay. Fe-NTA/H2O2 also induced fragmentation of bovine serum albumin (BSA) in vitro and depleted cellular GSH levels in lymphocytes. BSA fragmentation and GSH depletion were dose dependently counteracted by FPP. EPR spin trapping studies demonstrated that antioxidant properties of FPP are related to both hydroxyl scavenging as well as iron chelating properties. PMID- 11062701 TI - Antineoplastic activity of a novel multimeric gemcitabine-monophosphate prodrug against thyroid cancer cells in vitro. AB - Gemcitabine (Gem) is a deoxycytidine analog that is effective against pancreatic cancer and other malignancies following conversion to the 5'-O-mono-, di- and tri phosphate forms. We evaluated the cytotoxicity of GemMP[10], a novel multimeric form of 2'-deoxy-2',2"-difluorocytidine-5'-O-monophosphate (gemcitabine monophosphate) against three thyroid carcinoma cell lines established from anaplastic (8505C), papillary (B-CPAP) and poorly-differentiated papillary (BHT 101) cancer. GemMP[10] decreased tumor cell growth at concentrations ranging from 1 to 50 nM. These concentrations were 5- to 10-fold lower than those required for inhibition of tumor cell growth by monomeric Gem. GemMP[10] cytotoxicity occurred via induction of apoptosis. Flow cytometric analysis of GemMP[10] treated cells revealed growth arrest in S-phase. Fas-antigen expression was increased in thyroid cancer cells treated with GemMP[10], whereas Fas-L and Bcl-2 expression were not significantly affected. These results demonstrated that GemMP[10] is a potent cytotoxic agent that serves to induce apoptosis in association with increased Fas expression in cultured thyroid cancer cell lines. PMID- 11062702 TI - Inhibitory effects of active substances isolated from Cassia garrettiana heartwood on tumor growth and lung metastasis in Lewis lung carcinoma-bearing mice (Part 2). AB - Previously, we reported that a methanol extract (500 mg/kg x 2/day) of the heartwood of Cassia garrettiana inhibited the tumor growth and metastasis to the lung in Lewis lung carcinoma (LLC)-bearing mice. Furthermore, we isolated the two active substances from the methanol extract of C. garrettiana and identified compound 1 as cassigarol A. In the present study, compound 2 was identified as 3, 3', 4, 5'-tetrahydroxy stilbene (piceatannol) based on the 1H-NMR spectral data and products formed by oxidation with potassium permanganate. We examined the active substance (compound 2, piceatannol) and its acetylated derivative on the tumor growth and lung metastasis in LLC-bearing and carcinectomized mice. Piceatannol (50 mg and 100 mg/kg x 2/day) did not affect the tumor growth, while piceatannol acetate (50 mg and 100 mg/kg x 2/day) significantly inhibited the tumor growth. Piceatannol and its derivative piceatannol acetate inhibited the metastasis to the lung dose-dependently in carcinectomized mice. Moreover, piceatannol and piceatannol acetate prolonged the survival time and increased the survival rate in carcinectomized mice. In addition, piceatannol inhibited the formation of capillary-like networks of human umbilical vein endothelial cells (HUVECs) at the concentrations of 10 to 100 microM, but its acetylated derivative did not. Therefore, it is suggested that the antimetastatic activities of piceatannol might be due to the inhibition of tube formation (angiogenesis) of HUVECs. PMID- 11062703 TI - Effect of interferon-alpha A/D in combination with the Japanese and Chinese traditional herbal medicine juzen-taiho-to on lung metastasis of murine renal cell carcinoma. AB - Several studies have shown that the Kampo medicine Juzen-taiho-to (Si-Quan-Da-Bu Tang in Chinese) has various biological activities, including anti-tumor effects when combined with surgical excision or with chemotherapeutic drugs. Here we investigated the effect of combined therapy with interferon (IFN)-alpha A/D and Juzen-taiho-to on experimental lung metastasis of murine renal cell carcinoma (Renca) cells. Five consecutive administrations of IFN-alpha A/D to Renca-bearing mice resulted in dose-dependent inhibition of lung metastasis. IFN-alpha A/D at the dose of 100,000 IU/mouse significantly inhibited the metastasis, but a marked loss of body weight was observed during and after the administration. In contrast, oral administration of Juzen-taiho-to (50 mg/mouse) alone tended to inhibit the metastasis, but the effect was not statistically significant. The combination treatment of suboptimal doses of IFN-alpha A/D and Juzen-taiho-to markedly augmented the antimetastatic effect without causing any loss of body weight, as compared with either treatment alone. Similar results were also obtained by treatment with IFN-gamma in combination with Juzen-taiho-to. Clinically, immunotherapy with IFNs has been primarily approved for the treatment of patients with metastatic renal cell carcinoma, but sufficient efficacy has not yet been obtained. Therefore, the combination of IFNs with Juzen-taiho-to may provide a means to increase the therapeutic potential of IFNs and to decrease their toxicity for the treatment of metastatic renal cell carcinoma. PMID- 11062704 TI - Increase of tumour infiltrating lymphocytes in mice treated with antimetastatic doses of NAMI-A. AB - NAMI-A is a novel antitumour agent, based on ruthenium, which has proved effectiveness against lung metastases of solid mouse tumours. The study focuses on the effects of NAMI-A on leukocyte infiltration into the primary tumour of MCa mammary carcinoma, implanted subcutaneously (s.c.) or intramuscularly (i.m.) into CBA mice. NAMI-A, given with a cycle of daily treatments for six consecutive days on advanced tumours at 35 mg/kg/day, markedly reduces lung metastasis independently of the tumour type (Lewis lung carcinoma, MCa mammary carcinoma or TS/A adenocarcinoma) being treated and of the site of tumour implantation (s.c. or i.m.). The analysis of leukocyte infiltration of the primary tumour, performed on a single cell suspension of cells isolated from a Ficoll gradient on which a raw suspension of primary tumour cells was layered, showed NAMI-A to significantly increase tumour infiltrating lymphocytes. These lymphocytes are almost all CD3+ cells with a significant increase of the CD8+ over the CD4+ subpopulation that reduces the helper/suppressor ratio from 2.8 to 2.1. These data indicated the absence of toxicity of NAMI-A for tumour infiltrating lymphocytes and suggested that this compound might even synergize in combined treatments with cancer immunotherapy. PMID- 11062705 TI - The transcription factor ATF-2 inhibits extracellular signal regulated kinase expression and proliferation of human cancer cells. AB - The mitogen activated protein kinase (MAPK) pathway is a paradigm for regulation of growth factor signaling and cellular proliferation. The MAPK pathway is a major target for signaling by growth factor receptor kinases. The MAPK pathway consists of a series of protein kinases which is activated by phosphorylation of specific amino acid residues in their regulatory domains. The MAPK family can be divided into three subgroups: the extracellular signal regulated kinases (ERKs), the stress activated protein kinase/jun N terminal kinase (SAPK/JNK), and the p38 MAPK. These kinase cascades phosphorylate transcription factor targets such as ets, c-jun, and ATF-2. Of these, little is known about the role of ATF-2 in regulation of MAPK signaling and cellular proliferation. To begin to understand this role, we overexpressed ATF-2 in a human cancer cell line. ATF-2 inhibited the G1/S phase transition of the cell cycle and decreased the proliferation rate of these cells. Decreased proliferation correlated with cell cycle independent inhibition of ERK1 expression in ATF-2 clones. Genetic and pharmacologic inhibition of ERK1 activity was sufficient to reproduce the effects of ATF-2 on cell cycle progression and proliferation. These results indicate a novel role for ATF-2 in cancer cell proliferation and suggest a potential feedback mechanism that regulates MAPK signaling. PMID- 11062706 TI - Effects of anticancer drugs, metals and antioxidants on cytotoxic activity of benzothiepins/benzoxepins. AB - Among 11 benzothiepins/benzoxepins, 4-chloro-3,4-dihydro-2-(2-oxo-2-phenylethyl) 1-benzothiepin-5-(2H)-one [1] showed the highest cytotoxicity against human oral squamous cell carcinoma HSC-2 cells, followed by 2,3-dihydro-2-(2-oxopropyl)-2 phenyl-1-benzoxepin [2]. Popular antioxidants, such as N-acetyl-L-cysteine and sodium ascorbate significantly reduced the cytotoxic activity of [1] but not that of [2]. Compound [1] induced internucleosomal DNA fragmentation in human promyelocytic leukemic HL-60 cell line, but produced large DNA fragmentation in human oral tumor cell lines (HSC-2, HSG). Compounds [1] and doxorubicin additively reduced the viable cell number of HSC-2 cells. These data, taken together with their tumor specific action, demonstrate for the first time, the medicinal efficacy of benzothiepins/benzoxepins. PMID- 11062707 TI - Radical modulating activity and cytotoxic activity of synthesized eugenol-related compounds. AB - The ability of nine synthetic eugenol-related compounds to scavenge O2- (generated by the hypoxanthine-xanthine oxidase reaction) was compared with their radical generation and cytotoxic activity. ESR spectroscopy showed that eugenol (4-allyl-2-methoxyphenol), 2-allyl-4-methoxyphenol, 2-allyl-4-t-butylphenol and 2,4-dimethoxyphenol efficiently scavenged O2- and produced radicals under alkaline conditions. 2-allyl-4-t-butylphenol showed the highest cytotoxic activity and DNA-synthesis inhibitory activity, possibly due to the hydrophobic radical reactivity. 2-allyl-4-methoxyphenol and 2,4-dimethoxyphenol showed higher antioxidant activity than 3-t-butyl-4-hydroxyanisol (BHA), but all these compounds showed comparable cytotoxic activity with each other. These findings suggest a possible link between the cytotoxic activity and radical generation/scavenging activity in eugenol-related compounds. PMID- 11062708 TI - Potentiation of fludarabine cytotoxicity on non-Hodgkin's lymphoma by pentoxifylline and rituximab. AB - BACKGROUND: Fludarabine has become a drug of prominent use in hematopoietic malignancies exhibiting indolent growth profiles. Some studies show, however, that nearly 50% of patients are fludarabine-resistant from the very onset of therapy. Others relapse after an initial response rarely lasting more than 2 years. For this reason, modulating agents have been considered for use with fludarabine to potentiate fludarabine cytotoxicity and circumvent drug resistance. The chimeric anti-CD20 monoclonal antibody, Rituximab (IDEC-C2B8), is used to treat patients with low grade and follicular B-cell non-Hodgkin's lymphoma. Rituximab is known to inhibit the cell progression of tumor B cells and to sensitize them to chemotherapeutic drugs. A slower cell progression may enhance the efficacy of fludarabine incorporation, thus increasing its cytotoxicity. Therefore, the use of fludarabine and Rituximab in combination could potentiate fludarabine cytotoxicity. The methylxanthine, pentoxifylline, disrupts DNA repair mechanisms within a cell by not allowing a cell to arrest at the G2/M checkpoint. By not allowing cells to repair fludarabine DNA incorporation, pentoxifylline was thought to increase fludarabine-induced cytotoxicity in tumor cells. We tested these hypotheses in vitro. MATERIALS AND METHODS: Analysis of cytotoxicity was performed using the XTT assay on tumor cell lines and patient samples. RESULTS: Tumor cell models, including two B-cell non Hodgkin's lymphoma cell lines and a T-cell leukemia cell line, were shown to respond more effectively to fludarabine therapy in the presence of Rituximab or pentoxifylline. Two freshly derived B-cell chronic lymphocytic leukemia patient samples were also seen to exhibit a better response with a combination of fludarabine and pentoxifylline than with either alone. CONCLUSION: This study proves the hypothesis that Rituximab and pentoxifylline may provide a clinical approach to hinder the outgrowth of drug refractory tumor cells and achieve a longer period of remission. PMID- 11062709 TI - Mutation analysis of the cationic trypsinogen gene in patients with pancreatic cancer. AB - Recently, an Arg to His mutation at residue 117 of the cationic trypsinogen gene (Arg117His) has been shown to be associated with hereditary pancreatitis (hp). A serious complication of hp is development of pancreatic cancer. Patients suffering from hp have been reported to have a 53-fold increased risk to die from pancreatic cancer. However, the quantitative contribution of mutations in the cationic trypsinogen gene to all pancreatic cancer cases is unknown. A relevant contribution of the Arg117His-mutation to pathogenesis of pancreatic cancer might be possible, since also asymptomatic individuals have been reported to carry this mutation and individuals with only mild symptoms may be undiagnosed as hp. In the present study we analyzed genomic DNA obtained from pancreatic cancer tissue from 34 patients and corresponding normal tissue from 28 of these individuals. The third exon of the cationic trypsinogen gene was amplified by nested PCR and digested with AflIII, since the Arg117His mutation creates an AflIII-restriction site. None of the examined samples carried the Arg117His mutation, whereas the amplification product obtained from a patient with known hp was clearly positive. Sequencing of the complete third exon of the cationic trypsinogen gene in 10 of the pancreatic cancer patients resulted exclusively in the wild-type sequence. In addition DNA obtained from venous blood of 116 further patients with pancreatic cancer did not carry the Arg117His mutation. Our results show that the Arg117His mutation does not contribute to pathogenesis of a substantial fraction of all pancreatic adenocarcinomas. In contrast to most oncogenes or tumor suppressor genes the cationic trypsinogen gene (3rd exon) does not contain mutational hot spots. PMID- 11062710 TI - Morphological and phenotypic characterization of a new established ovarian carcinoma cell line (OvBH-1). AB - BACKGROUND: A limited number of permanent ovarian carcinoma cell lines have been described so far and the majority of them have been derived from ovarian ascitic fluid cells taken from patients with serous ovarian carcinoma usually after chemotherapy treatment. MATERIALS AND METHODS: The cells for culture were obtained from ascitic fluid cells of a patient with ovarian clear cell carcinoma. Cytomorphological analysis of cultured cells at early and late passages was performed by hematoxylin-eosin staining. Immunophenotypic characterization of cells was performed using the following monoclonal antibodies against: intermediated cellular filaments (CK 6/18, CK 7, CK 1,5,6,8,10,14,18, V9) ovarian carcinoma-associated antigens (OC125, OV-TL3), carcinoembryonic antigen, p53 and c-erbB-2 oncoproteins. RESULTS: In the established ovarian carcinoma cell line (OvBH-1) two morphologically distinct cell types were recognized. Cytomorphologically the dominating type appears to frankly malignant features. The second cell subtype showed a lower degree of malignant features. The epithelial origin of both cell types was confirmed by immunohistochemical staining using antibodies against different cytokeratin epitopes. The expression of tumor-associated antigens (CA125, OV-TL3) was found in both cell subtypes reflecting their origin from ovarian carcinoma. The cell line was negative for CEA staining. The genetic defects of cultured cells were revealed by detection of p53 and c-erbB-2 overexpression. The level of both oncoproteins and especially c erbB-2 was higher in the cell subtype with frankly malignant morphological features. CONCLUSIONS: A new established, well characterized ovarian clear cell carcinoma line OvBH-1 provides an experimental model for further investigation of the biological alterations responsible for carcinogenesis and chemoresistance of this uncommon subtype of epithelial ovarian carcinomas. PMID- 11062711 TI - Lymphatic microvessel density in epithelial ovarian cancer: its impact on prognosis. AB - BACKGROUND: The influence of lymphatic microvessel density (MVD) on survival in epithelial ovarian cancer is still unknown, owing to the fact that until recently no reliable immunohistologic markers for lymphatic endothelium were available. MATERIALS AND METHODS: By using a polyclonal antibody staining podoplanin, a novel marker for lymphatic endothelium, lymphatic MVD in tissue samples of 90 patients with epithelial ovarian cancer treated by radical surgery and chemotherapy was investigated. Survival analysis was performed using univariate and multivariate analysis. Furthermore, lymphatic MVD was compared to MVD assessed by CD34 immunostaining. RESULTS: Lymphatic MVD was significantly lower than CD34 MVD (p < 0.0001). There was no significant association between lymphatic MVD and various histological and clinical parameters. Lymphatic MVD had no influence on overall survival and disease free survival (p = 0.4627 and p = 0.4337, respectively; log-rank test). CONCLUSION: The formation of lymphatic vessels has no influence on the progression of epithelial ovarian cancer. PMID- 11062712 TI - Intracellular expression of IL-4 and inhibition of IFN-gamma by extracts from European mistletoe is related to induction of apoptosis. AB - BACKGROUND: Extracts from European mistletoe are used for adjuvant cancer treatment. Their influence on the intracellular expression of cytokines of the T helper cells type-1 (Th1; IFN-gamma) or type-2 (Th2; IL-4) is still unknown. MATERIALS AND METHODS: Lymphocytes from controls were incubated with mistletoe extracts (ME) and mistletoe lectins (ML) for 24 hours and co-stimulated with PMA/Ca-ionophore/monensin during the last 6 hours. Apoptosis and intracellular cytokine expression were detected by flow cytometry, the cytokine release into the supernatants by ELISA. RESULTS: ME and ML significantly inhibited intracellular expression of IFN-gamma but stimulated IL-4. Thereby, IL-4 was mainly expressed in apoptotic (Apo2.7+) cells. However, IFN-gamma secretion into the supernatants of the cells was dose-dependently inhibited by ME and ML, while IL-4 was not detected at all. CONCLUSION: The intracellular expression of the 'Th2-cytokine' IL-4 in ME- and ML-exposed cells may not be related to a typical Th2-response but rather to cell death. This effect might be of great relevance e.g. after intratumoural injection of the mistletoe extracts and, in general, for the inhibition of an inflammatory response during apoptosis. PMID- 11062713 TI - Cytostatic drugs and health risks for exposed personnel: search for new biomarkers. AB - The use of antiblastic drugs has opened up new perspectives in improvement of therapy and life quality for cancer patients. The widespread clinical application of cytostatic drugs implies risks for exposed hospital personnel, due to genotoxic and toxic-reproductive effects. Biological monitoring is fundamental to identify individuals at risk but is limited by the long latency of chronic effects, absence of unique cellular targets and low sensitivity of available laboratory tests. The objective of this study was to investigate toxic mechanisms by a molecular biology approach, searching for biomarkers potentially useful in monitoring programs. The proposed experimental model consisted of cell line exposure to cyclophosphamide, an alkylating agent of wide clinical use. Cellular response has been investigated focusing on potential targets at RNA level, through reverse transcription polymerase chain reaction (RT-PCR) and differential display analysis. We studied the expression of several genes involved in differentiation, apoptosis and chemoresistance: ets1, bax, bcl-2, bag-1, bcl-X, mdr1 and mrp. Specific patterns of mRNA modulations were observed. Differential display analysis revealed candidate genes induced or repressed following exposure: their characterization is in progress. Besides improving the understanding of toxic mechanisms, identification of modulated molecular targets opens up new perspectives in exposure risk assessment, biomonitoring and preventive strategies at occupational level. PMID- 11062714 TI - In vitro activity of human chorionic gonadotropin (hCG)--doxorubicin conjugates against ovarian cancer cells. AB - BACKGROUND: Despite radical surgery and the introduction of novel chemotherapeutic agents, the prognosis of ovarian cancer remains poor. Since in the past the potential role of gonadotropins in the induction and progression of ovarian cancer has been discussed, we conjugated doxorubicin to human chorionic gonadotropin (hCG) in order to specifically target ovarian cancer cells. MATERIALS AND METHODS: Doxorubicin was conjugated to hCG via glutaraldehyde. 48 hours post seeding, NIH:OVCAR 3 cells were treated with various concentrations of hCG-conjugated and non-conjugated doxorubicin for 2 hours. Cells were cultured for a total of 168 hours. Cell growth was monitored by a crystal violet assay. RESULTS: Conjugating doxorubicin to hCG resulted in an average specific uptake of 22 mol doxorubicin per mol protein (range 3.0-43.3 mol). Incubating NIH:OVCAR 3 cells for 2 hours with the conjugate led to a more than 8 fold increase of the IC50 values compared to non-conjugated doxorubicin (0.55 microM versus 4.43 microM). The antiproliferative activity of both conjugated and free doxorubicin was detectable up to 168 hours post treatment. CONCLUSIONS: The present experiments clearly demonstrate a more than 8-fold increase in cytotoxicity of the conjugates compared to free doxorubicin. It was also shown that this effect was not restricted to an acute toxic event but that it resulted in a prolonged antiproliferative activity. PMID- 11062715 TI - Iron- and aluminum-induced carcinogenesis. AB - Ferric and aluminum complexes with ATP have shown the induction of tumors in the site of subcutaneous injection, whereas sodium ATP has not. A concomitant but apparently independent phenomenon was a severe lymphoadenitis. The tumor calcium concentration showed an inverse relationship with the tumor growth rate. Carcinogenesis and lymphoadenitis are discussed considering well known effects of ferric and aluminum complexes with ATP on the cellular calcium homeostasis and of ATP on lymphatic tissue proliferation. PMID- 11062716 TI - Binuclear cells induced by acridine orange in giant cell tumor of bone. AB - We have recently found the presence of many binuclear cells among isolated and smeared cells in giant cell tumor of the bone (GCT). These binuclear cells are possibly associated with the formation of multinuclear cells. Therefore, this study was undertaken to clarify the mechanism of binucleation in GCT, using primary culture cells exposed to acridine orange (AO) which is a fluorescent vital staining dye for the cytoplasm and nucleus and which inhibits mitosis. The cells were isolated from explants of fresh tumor materials obtained from two GCT patients (GCT1 and GCT2). These cells were cultured in Dulbecco's modified Eagle medium (DMEM) with 10% Fetal calf serum (FCS). After exposure to 0.5 microgram/ml AO, for 0, 6, 24, 48, 96 and 144 hours the following parameters were investigated: 1) cell growth rate (GR); 2) frequency of hyperdiploid cells (%HDC) by DNA cytofluorometry; 3) mitotic index (MI); 4) BrdU labeling index (LI); 5) frequency of binuclear cells (%BNC). Compared to the control cells which were cultured in AO-free medium, the GR of both GCT cells exposed to AO was remarkably inhibited. The MI was 0 from 24 to 144 hours. The %HDC was increased at 24 hours and was maintained high until 144 hours. The LI was temporarily increased at 6 hours, but was decreased at 48 hours. The %BNC was gradually increased. AO inhibited DNA synthesis and cell mitotic activity in cultured GCT cells and it finally caused inhibition of cell growth. However, the frequencies of G2 arrest cells and binuclear cells were increased. These results suggested that the binuclear cells in GCT may be formed from G2 arrest cells by amitotic nuclear division, but not by mitosis without cytoplasmic division, or by cell fusion. PMID- 11062717 TI - Fluorovisualization effect of acridine orange on mouse osteosarcoma. AB - If the localization of musculoskeletal sarcomas could be visualized during surgery, it would be possible to completely resect the tumor with minimum damage to normal tissues and the patients could retain a functional limb. Therefore, we conducted the present study to clarify the usefulness of acridine orange (AO) for fluorovisualization of tumors using a mouse osteosarcoma model. At 2 hours after injection of 10 mg/kg AO to mice inoculated with MOS mouse osteosarcoma cells, fluorovisualization of mouse osteosarcoma reached the maximum level. Even a 1-mm diameter lesion of pulmonary metastasis was visualized. The results suggested that AO may be useful for specific fluorovisualization of human osteosarcomas during surgery. PMID- 11062718 TI - Significance of tumor associated tissue eosinophilia and other inflammatory cell infiltrate in early esophageal squamous cell carcinoma. AB - Tumor associated inflammatory cell infiltration plays an important role in the biological behavior of cancer as one of the carcinoma-stromal interactions. In this study, we characterized inflammatory cell infiltration on esophageal squamous cell carcinoma (SqCC) and correlated the findings with various clinicopathological factors, including clinical outcome of the patients, in order to study its biological significance. We examined 35 cases of surgically resected early esophageal SqCC or carcinoma with invasion limited to the submucosal layer. We evaluated the abundance of CD4+ T-cell, CD8+ T-cell, B-cell, plasma cell, CD68+ macrophage, neutrophil and eosinophil in the stroma adjacent to the tumor and correlated these findings with clinicopathological factors. The cases without LN metastasis had a significantly larger number of tumor associated eosinophils than those with LN metastasis. Primary lesions in cases without LN metastasis tended to demonstrate more CD68+ macrophage infiltration than those with LN metastasis, but the difference did not reach statistical significance. In addition, the cases with more than 50 eosinophils and macrophages per 10 high power field in the primary lesion demonstrated a significantly smaller incidence of LN metastasis than those with less than 50 eosinophils and macrophages per 10 high power field. Thus tumor associated tissue eosinophila, also known as TATE, is considered to be involved in the biological behaviour of early esophageal SqCC, especially in their metastatic potential. PMID- 11062719 TI - Prognostic relevance of urokinase type plasminogen activator, its receptor and inhibitors in chondrosarcoma. AB - The plasminogen activation system plays an important role in enhancing pericellular proteolysis of tumor invasion/metastasis and in autocrine/paracrine tumor growth stimulation. To investigate the prognostic significance of the plasminogen activation system in human chondrosarcoma, the immunohistochemical expression of urokinase-type plasminogen activator (uPA), urokinase-type plasminogen activator receptor (uPAR), plasminogen activator inhibitor, 1 (PAI-1) and 2 (PAI-2) were analyzed in 28 patients with chondrosarcoma. In multivariate survival analysis, histological grade (p = 0.0008) and location (p = 0.02) were independent risk factors for local relapse. For metastasis-free survival, uPA index (p = 0.006) and PAI-2 index (p = 0.04) were independent prognostic factors. PAI-2 index (p = 0.02), uPAR index (p = 0.02) and histological grade (p = 0.03) predicted total survival. These results demonstrated the usefulness of uPA, uPAR and PAI-2 expression as biological prognostic indicator and the importance of the plasminogen activation system in tumor progression and metastasis in chondrosarcoma. PMID- 11062720 TI - Spatial expression of a DNA repair gene, N-methylpurine-DNA glycosylase (MPG) during development in mice. AB - BACKGROUND: DNA repair is a crucial phenomenon that maintains the chromosome integrity of genome which are continuously damaged by endogenous and exogenous alkylating agents. If the damaged DNA is not repaired, it may lead to mutation, chromosomal aberration, aging and cancer. N-methylpurine-DNA glycosylase (MPG), a ubiquitous DNA repair enzyme, removes N-methylpurine and other damaged purines in DNA. MATERIALS AND METHODS: MPG mRNA expression was revealed at various stages of mouse development from day 7.5 p.c. (post coitum) embryo to day 400 mature adult by Northern blot hybridization or RT-PCR. RESULTS: MPG transcripts were abundant in the mouse embryo during pregnancy and in adult testis and ovary. The MPG mRNA level in the testis was low in 1-week-old mice, but the level showed its maximum among the organs tested in 4-week-old young adults. In placenta, the level of MPG mRNA continuously decreased from day 7.5 p.c. to day 17.5 p.c. CONCLUSIONS: The spatial expression of MPG gene is highly regulated. Transcription of MPG is maximum in rapidly dividing and growing tissues during development. These data suggest that an elevated rate of MPG transcription is required for DNA replication. PMID- 11062721 TI - Serum levels of matrix metalloproteinase-2 and -9 in patients with head and neck squamous cell carcinoma. AB - BACKGROUND: Matrix Metalloproteinases (MMPs) have been implicated as playing an important role in cancer invasion and metastasis. MMPs have been identified in a wide variety of malignancies including head and neck squamous cell carcinomas (HNSCC). MATERIAL AND METHODS: We investigated the circulating level of MMP-2 (gelatinase A or 72-kD type IV collagenase) and MMP-9 (gelatinase B or 92-kD type IV collagenase) in sera from patients with various head and neck squamous cell carcinomas (n = 86) as well as from healthy normal controls (n = 47). Serum MMP concentrations were determined as serum immunoreactivity by using a quantitative sandwich enzyme immunoassay technique. For statistical analysis, the t-test and Kruskal-Wallis test were performed. RESULTS: The majority of the patients with HNSCC were found to have high concentrations of serum MMP-9. The levels of MMP-9 in the sera of patients with cancer ranged from 39 to 1547 ng/ml (mean, 417 ng/ml). In contrast, the MMP-9 serum levels in 47 healthy individuals ranged from 30 to 537 ng/ml (mean, 189 ng/ml), MMP-9 serum concentration being significantly higher in HNSCC patients (p = 0.001). MMP-9 serum concentrations of patients with advanced stage HNSCC were significantly higher (p = 0.0449) compared to patients with early stage cancer. No significant difference of MMP-2 serum levels was seen when comparing HNSCC patients and normal controls. CONCLUSION: The present data indicate that the elevation of serum levels of MMP-9, but not MMP-2, may be a useful marker for clinical monitoring of HNSCC patients. PMID- 11062722 TI - Expression and activity of DNA-dependent protein kinase in normal human leukocytes. AB - BACKGROUND: DNA-PK is a serine/threonine protein kinase that has been shown to play an important role in V(D)J recombination, Ig switching and DNA double strand break (DSB) repair. We have investigated DNA-PK in whole cell extracts from B and T lymphocytes, monocytes and granulocytes and the effects of phorbolmyristate acetate (PMA) and Concanavalin A (Con A) on DNA-PK in B and T lymphocytes. MATERIALS AND METHODS: Lymphocytes were separated from human peripheral blood using the E-rosette technique and DNA-PK was analysed using Western blot, DNA-PK kinase activity assay and flow cytometry. RESULTS: The levels of DNA-PK activity and content varied among the leukocytes. PMA and Con A stimulation consistently increased both DNA-PK activity and content in B lymphocytes as cells entered the S phase. Up-regulation could normally not be seen in T lymphocytes, although cells entered the S phase following PMA and Con A stimulation. CONCLUSION: Mitogenic stimulation up-regulates DNA-PK activity and protein expression in B lymphocytes. T lymphocytes show interindividual differences in DNA-PK. Regulation of DNA-PK is not strictly cell cycle dependent for all cell types. PMID- 11062723 TI - In vitro effects of fenretinide on cell-matrix interactions. AB - BACKGROUND: Understanding the molecular basis of the metastatic spread of cancer and the underlying mechanisms is crucial for the development and appropriate clinical use of novel therapeutic agents directed at prevention of metastasis. Retinoids have been reported to inhibit cell proliferation, modulate cell differentiation, enhance apoptosis and to prevent the conversion of in situ cancer to locally invasive malignancy by suppressing the invasive process as well as by inhibiting angiogenesis. Fenretinide (4-HPR), a synthetic derivative of retinoic acid, is less toxic than natural retinoids and is active in the prevention and treatment of a variety of tumours in animal models. Its efficacy in cancer chemoprevention and therapy has been investigated in clinical trials. MATERIALS AND METHODS: In order to evaluate the effects of 4-HPR on the late stages of tumour progression, chemically transformed BALB/c 3T3 cells, showing a fully malignant phenotype, were exposed to 4-HPR (0.25-10 microM; 72 hours pre treatment) and then analysed for in vitro invasive ability. The possible mechanisms of action responsible for the anti-invasive activity of 4-HPR were investigated, analysing cellular adhesion, motility, and proteolytic capability. RESULTS: Data showed that 4-HPR significantly inhibited the invasive phenotype of chemically transformed cells; the reduction in Matrigel invasion was dose dependent and seemed not to be related to cytotoxic effects or reduction in cell proliferation rates induced by 4-HPR assayed doses. The 4-HPR-induced decrease in chemotactic motility of transformed cells correlated well with the invasion inhibition. 4-HPR, at active concentrations, differently affected cell adhesion to the extracellular matrix, depending on the coating substrate used (laminin, collagen IV, fibronectin and vitronectin). 4-HPR treatment significantly enhanced cell adhesion to laminin, while reducing cell-vitronectin attachment. It did not modify the attachment of the cells to fibronectin and collagen IV. Zymographic analysis failed to demonstrate 4-HPR involvement in the modulation of the activity and expression of gelatine degrading enzymes. CONCLUSION: These data suggest that 4-HPR inhibits tumour cell invasion through a basement-like matrix, by suppressing chemotactic motility and by altering cell-matrix interactions. PMID- 11062724 TI - Adamantyl maleimide induced changes in adhesion molecules and ROS are involved in apoptosis of human gastric cancer cells. AB - BACKGROUND: We have previously found that N-1-adamantyl maleimide (AMI) inhibited the growth of SC-M1 tumors in vitro and in vivo. The cytotoxicity of AMI on SC-M1 was accompanied by a decrease of adherent cells and the suppressive effect was associated with conformational changes in cell membrane protein. In order to determine the cellular targets of AMI in human gastric cancer SC-M1 cells, we examined AMI-induced changes in the levels of adhesion molecules CD29 (beta 1 integrin) and CD54 (ICAM-1) and GSH. In addition, we also analyzed changes of apoptosis markers such as annexin V binding to membrane and caspase 3 activity in SC-M1 cells after treatment with AMI. MATERIALS AND METHODS: Changes in CD29, CD54, annexin V binding and GSH levels were examined using FITC-conjugated antibodies or fluorescence probes and flowcytometry. Caspase 3 activity was assayed with spectrofluorometry. RESULTS: We found that the expression of CD29 and CD54 on SC-M1 was decreased and the caspase 3 activity was increased during the early apoptosis induced by AMI. Moreover, it was found that the GSH content of the cell was depleted within 30 minutes and then recovered. CONCLUSION: These results suggest that the cell membrane proteins, such as adhesion molecules (CD29, CD54) and intracellular GSH, were the targets of AMI on SC-M1 cells. Since these membrane alterations were prior to apoptosis they may have transduced a death signal to SC-M1 cells. PMID- 11062725 TI - p27Kip1 is the key mediator of phenylacetate induced cell cycle arrest in human prostate cancer cells. AB - Treatment with millimolar concentrations of phenylacetate (PA), results in cytostasis, growth inhibition and differentiation in several human cancer cell lines, including prostate cancer. However, the molecular basis of PA-induced biological effects has not been elucidated in detail. In this study we focused on its influence on cell cycle events and investigated alterations in cell cycle regulators in androgen-dependent and independent human prostate cancer cell lines. FACS analysis revealed that suppression of cell growth by PA was due to G1 arrest, with reduced phosphorylation of the retinoblastoma protein (pRb) and CDK2 activity. Expression of p27Kip1 was increased, while p21Cip1, p53, cyclinD1 and cyclin E were not affected by PA. Binding of p27Kip1 to CDK2 increased significantly following treatment with PA. Furthermore, antisense p27Kip1 oligonucleotide attenuated the inhibitory effect of PA. Our results suggested that p27Kip1 might be a critical target in PA-mediated cell growth arrest in prostate cancer cells playing a key role in CDK2 inactivation followed by hypophosphorylation of pRB and subsequent G1 cell cycle arrest. PMID- 11062726 TI - Does human melanoma express carcinoembryonic antigen? AB - BACKGROUND: Several investigators have proposed that carcinoembryonic antigen (CEA), an immunogenic antigen expressed by colon carcinoma, may also be expressed by human melanoma. Because sialyl Lewisx (sLex), the carbohydrate moiety of CEA, has been identified in melanoma, we compared CEA and sLex levels in colon carcinoma cells and melanoma cells. METHODS: CEA levels were assessed for expression on the cell surface and in cell lysates of cutaneous melanoma cell lines by two different kinds of ELISA, and by Western blot analysis of immunoprecipitated CEA using monoclonal antibodies (Mabs) T84-66 and COL-1, which have defined specificities for CEA. Colon carcinoma cells and purified CEA were positive controls. RESULTS: Both Mabs reacted strongly with cell surface and cell lysates of colon cancer. Mab T84-66 reacted well with cell surface but not cell lysates of melanoma. COL-1 reacted poorly with cell surface but its binding increased with the density of melanoma cell lysates. Both Mabs intensely stained the blots of purified CEA and colon carcinoma lysates immunoprecipitated with the respective Mabs, but failed to stain the immunoprecipitates of melanoma cell lysates. Both Mabs bound to lysates immunoprecipitated with anti-sLex Mab in colon carcinoma, but not in melanoma. Cell-surface expression of CEA and sLex was significantly correlated (r2: 0.88) in colon cancer cells but not in melanoma. CONCLUSION: Our results confirm the presence of CEA in colon carcinoma but not in human cutaneous melanoma cell lines. PMID- 11062727 TI - Binding of tumor antigen mucin (MUC1) derived peptides to the heat shock protein DnaK. AB - The human epithelial mucin encoded by the gene MUC1 is a tumor-associated antigen expressed on breast, pancreatic, colon and ovarian cancer cells recognized by cytotoxic T-cells and antibodies. Underglycosylated as well as glycosylated mucin peptide epitopes are promising targets for vaccination against cancer. Heat shock proteins of 70 kDa (HSP70), also highly expressed in tumor cells, can function as chaperones for peptides and proteins and are involved in antigen processing. The involvement of HSP70 molecules in mucin antigen binding, processing and presentation has not yet been examined. Here we present first results concerning the relative binding affinities of various mucin-derived peptides to the bacterial 70 kDa heat shock protein DnaK. Interestingly, longer mucin peptides reveal a higher affinity to DnaK than short peptides. The non-glycosylated mucin derived peptides of 5-8 amino acids length were able to compete with a high affinity (unrelated) reference peptide at millimolar concentrations. Glycosylation of the investigated short peptides lowers their binding affinity to DnaK, depending on the position of the carbohydrate. The binding affinity is not influenced by free charges at unprotected ends. The peptide (MUC1)5 consisting of five repeating units has an affinity enhanced by a factor of three as compared to the peptide with only one repeating unit. Mucin-peptide-HSP-complexes could be the basis of developing new kinds of tumor vaccines. PMID- 11062728 TI - Measurement of sister chromatid exchange induction in intracerebral brain tumors as a method for evaluation of therapeutic drug combinations. AB - Rats with 9L or 9L-2 intracerebral brain tumors were treated with streptozotocin (STZ) and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) at various doses either as single agents or in combination. Treatment of rats bearing 9L or 9L-2 tumors with either STZ or BCNU produced a significant increase the level of sister chromatid exchanges (SCEs) (p < .001). Compared with 9L-2 tumors, 9L tumors were 7.8-fold more sensitive to the induction of SCEs by BCNU treatment. After combination treatments of STZ and BCNU, the number of SCEs observed in 9L tumors was additive but was synergistic in 9L-2 tumors. O6-alkylguanine DNA alkyltransferase activity (AGT) was measured in 9L and 9L-2 cells in vitro. No AGT activity was detected in 9L cells, however, a low level of activity was measured in 9L-2. In vitro, treatment of 9L-2 cells with STZ produced a dose dependent inhibition of AGT activity. We interpret, these results to suggest that STZ pretreatment potentiated the effects of BCNU in 9L-2 tumors by inhibiting AGT activity. The observations in this study suggest that measurement of SCE induction in intracerebral tumor models may provide a useful strategy for evaluating the effectiveness of new agents and potential therapeutic combinations for the treatment of gliomas. PMID- 11062729 TI - Concentration-dependent variable effects of etoposide on the cell cycle of CML cells. AB - BACKGROUND: Etoposide, a DNA-topoisomerase II inhibitor, is used for a broad spectrum of cancers with various therapeutic strategies. But the molecular mechanisms of its concentration-dependent effects are not clearly defined. MATERIALS AND METHODS: Chronic myelogenous leukemia K562 cells were treated with low (5 microM) or high (100 microM) concentrations of this drug and the changes of cell cycle progression, expression of cell cycle regulating genes and cyclin B1-dependent histone H1 kinase activity were studied. RESULTS: In the presence of 5 microM etoposide, K562 cells continued to synthesize DNA and most cells showed progress into G2 phase until 24 hours. In contrast, 100 microM etoposide rapidly inhibited DNA synthesis by around 6 hours and most cells remained in their initial phase, while the incorporation of bromodeoxyuridine was partially resumed from 12 hours. The histone H1 kinase activity was only down-regulated in the early phase of 100 microM treated cells. Among the cell cycle controlling genes, c-Myc and P21Cip1/WAF1 showed impressive responses to the two etoposide concentrations. At 100 microM, c-Myc protein rapidly vanished at 3 hours, while p21Cip1/WAF1 was inversely induced from 3 hours. These changes were also observed at 5 microM, but they occurred slowly and weakly. CONCLUSION: The present findings indicate that two concentrations of etoposide functioned as an anticancer agent through modulating the genes related in cell cycle progression. Differing responses of c-Myc and p21Cip1/WAF1 at two concentrations may govern the antiproliferative effects of etoposide. PMID- 11062730 TI - Cachexia induction by EL-4 lymphoma in mice and possible involvement of impaired lipoprotein lipase activity. AB - Several lines of evidence have postulated that reduction in the activity of lipoprotein lipase (LPL) is involved in cachexia induction in cancer patients. Recently we have demonstrated that murine melanoma B16 has the ability to reduce the LPL activity and thereby induce cachexia symptoms in mice following intraperitoneal inoculation. In order to further investigate the relationship between LPL activity and cachectic syndrome, cachexia models other than melanoma B16 are required. However, there are few animal cachexia models in which LPL activity is involved in the induction of cachectic symptoms. In this study, cachectic symptoms and plasma LPL activity were investigated in mice bearing EL-4 mouse lymphoma. In EL-4 bearing mice the body weight including tumor weight in the abdominal cavity was rather higher than that of normal mice without tumor, whereas weights of carcass wet and gastrocnemius muscle were significantly decreased in EL-4 bearing mice. Elevated blood levels of triglyceride and non esterified fatty acid were observed in mice bearing EL-4, associated with the impaired plasma LPL activity. Overall, this study indicated that EL-4 lymphoma in mice results in a severe cachexia which is possibly related to impaired LPL activity and also provided a useful cachexia model for understanding the role of LPL in the development of cancer cachexia. PMID- 11062731 TI - Involvement of caspases in 5-FU induced apoptosis in an oral cancer cell line. AB - Although many anticancer drugs have been reported to induce apoptosis in cancer cells, the underlying mechanism remains unclear (1-3). Recent studies have revealed that the caspase family of cysteine proteases have been shown to play an important role in the regulation of several apoptotic processes. Thus, the present study investigated whether apoptosis induced by anticancer drugs is mediated by the activation of caspase cascade. NA cells, a squamous cell carcinoma cell line, were exposed to cisplatin (CDDP) or 5-fluorouracil (5-FU) with or without inhibitors of caspase 1, 3 and 8. Analysis of DNA fragmentation revealed that caspase inhibitors consistently inhibited DNA fragmentation induced by 5-FU. During the early stages of apoptosis, phosphatidylserine (PS) is translocated from the inner side of the plasma membrane to the cell surface. This PS externalization was markedly inhibited by treatment with caspase-8 inhibitor. These findings suggested that 5-FU induced apoptosis was mediated by the activation of a caspase cascade involving caspase 1, 3 and 8. PMID- 11062732 TI - Bcl-2-independent induction of apoptosis by neuropeptide receptor antagonist in human small cell lung carcinoma cells. AB - The broad-spectrum antagonist of neuropeptide receptor, [D-Arg1, D-Phe5, D Trp7,9, Leu11]substance P, induced apoptosis selectively in human small cell lung carcinoma (SCLC) cells, which express gastrin-releasing peptide receptor, but not in other types of tumor cells as well as normal cells. The addition of gastrin releasing peptide or bombesin and the inhibitor of caspase-3 suppressed [D-Arg1, D-Phe5, D-Trp7,9, Leu11]substance P-induced apoptosis. Moreover, [D-Arg1, D-Phe5, D-Trp7,9, Leu11]substance P-induced apoptosis was not suppressed by Bcl-2 over expression. Thus, blockage of gastrin-releasing peptide receptor-mediated signaling may provide a novel therapeutic option in SCLC which has become resistant to conventional chemotherapeutic agents. PMID- 11062733 TI - Synergistic effects of danazol and mifepristone on the cytotoxicity of UCN-01 in hormone-responsive breast cancer cells. AB - BACKGROUND: 7-Hydroxystaurosporine (UCN-01) is a novel antitumor agent as well as a potent inhibitor of a variety of protein kinases with a preference to protein kinase C. Because an intimate interaction exists between PKC signaling and the ER signaling pathways, sex steroid agonists/antagonists might modulate the cytotoxicity of UCN-01. MATERIALS AND METHODS: Effects of sex steroid agonists and antagonists on the UCN-01 cytotoxicity were analyzed by MTT assay in MCF-7/WT and MCF-7/ADR, a multiple drug resistance phenotype lacking ER and PR. RESULTS: MCF-7/ADR is 23-fold more resistant to UCN-01 than MCF-7. In MCF-7/WT, danazol and mifepristone enhanced the cytotoxicity of UCN-01, while these agents did not exert any significant effects on it in MCF-7/ADR. CONCLUSION: These results suggest that danazol or mifepristone might enhance the antitumor activity of UCN 01 in hormone-dependent cancer cells by interacting with PR. PMID- 11062734 TI - Reduced invasiveness and metastasis of Chinese hamster ovary cells transfected with human interleukin-17 gene. AB - Human IL-17 (hIL-17) stimulates epithelial, endothelial, fibroblastic cells and macrophages to secrete various cytokines. The present study was designed to assess the effects of the transfection of the hIL-17 gene in Chinese hamster ovary (CHO) cells. A complementary DNA (cDNA)-encoding hIL-17 was obtained by polymerase chain reaction (PCR) amplification from human CD4+ T-cell cDNA and inserted into the plasmid pRc/CMV to construct an expression vector for hIL-17. CHO cells were transduced with hIL-17 DNA-carrying cytomegalovirus (CMV)-based retroviral vectors. A clone with a high mRNA expression of hIL-17 (CHO/IL-17) was selected by Northem blotting. There was no significant difference in the in vitro growth of cells among parent CHO cells, vector-only transfected cells (CHO/neo) and CHO/IL-17 cells. A Matrigel invasion chamber assay, however, demonstrated significantly lower invasiveness by CHO/IL-17 cells than by either the parent CHO or the CHO/neo cells. There was no difference in the in vivo growth among the cells, when subcutaneously transplanted into nude mice. When injected into the tail vein, however, the number of metastatic nodules in the lungs of CHO/IL-17 injected mice was significantly smaller than that of CHO- or CHO/neo-injected mice. Furthermore, NK activity of spleen cells was significantly higher in nude mice transplanted with CHO/IL-17 cells than in mice transplanted with parent CHO or CHO/neo cells. In conclusion, the hIL-17-gene-transfected CHO cells showed a significantly lower metastatic potential to the lung by directly modulating the invasiveness and metastasis of CHO cells as well as by enhancing NK activity. PMID- 11062735 TI - Effect of cobalt ion on radical intensity and cytotoxic activity of antioxidants. AB - The effect of CoCl2 on the cytotoxic activity of various antioxidants against human oral tumor cell lines (HSC-2, HSG) and normal human gingival fibroblasts (HGF) was investigated. Noncytotoxic concentrations of CoCl2 significantly reduced the cytotoxic activity of sodium ascorbate, gallic acid, epigallocatechin gallate (EGCG), curcumin and dopamine, but not that of sodium 5,6-benzylidene-L ascorbate (SBA) and benzaldehyde. Among these compounds, benzaldehyde showed the most prominent tumor-specific cytotoxic action. ESR spectroscopy showed that these antioxidants produced radicals under alkaline condition and that their radical intensity was transiently enhanced and finally disappeared by addition of CoCl2. Antioxidants which are sensitive to CoCl2 generally had higher cytotoxic activity and oxidation potential (measured by NO monitor) and addition of CoCl2 significantly reduced their oxidation potential. The present study suggests that cobalt ion stimulates the oxidation of antioxidants to their inactive products. PMID- 11062736 TI - p53 and MDM2 protein expression in squamous cell carcinoma and neighboring dysplasia from the upper aerodigestive tract. AB - BACKGROUND: The identification of genetic changes in preinvasive epithelial lesions may provide markers for a better assessment of the progression potential. MATERIALS AND METHODS: p53, MDM2 and PCNA immunophenotypes were examined in 57 cases of squamous cell carcinoma (SCC) of the upper aerodigestive tract and the adjacent normal and dysplastic epithelia. RESULTS: PCNA index increased with increasing grade of dysplasia. p53 protein was expressed in 35% and MDM2 protein in 33% of SCCs. The p53 protein was expressed in 89% of mild and moderate and in 93% of severe dysplasia and carcinoma in situ adjacent to p53-positive SCCs. The MDM2 protein was expressed in 30% of mild and moderate and in 54% of severe dysplasia and carcinoma in situ adjacent to MDM2-positive SCCs. Preinvasive lesions adjacent to negative SCCs stained negative. CONCLUSIONS: p53 protein was detected more frequently in preinvasive lesions than MDM2 protein and seems to be of greater value as a biomarker. PMID- 11062737 TI - Everted rat intestinal sacs: a new model for the quantitation of P-glycoprotein mediated-efflux of anticancer agents. AB - P-Glycoprotein (p-gp) situated in the epithelium of the gastrointestinal tract is a recognised barrier limiting oral absorption of antitumour agents. Here we describe the rat everted gut sac as a new in vitro model for quantitation of p-gp mediated efflux of anti-cancer agents using [3H]vinblastine, [14C] doxorubicin and verapamil as reference compounds. Tissue and serosal uptake of [3H]vinblastine was linear over 90 min (0.031 +/- 0.001 and 0.003 +/- 0.001 ng/mg protein/h respectively). [14C]Doxorubicin tissue accumulation was significantly lower; 0.006 +/- 0.001 ng/mg protein/h (p < 0.05) whereas serosal transfer was significantly higher 0.017 +/- 0.001 ng/mg protein/h (p < 0.05) than [3H]vinblastine. Addition of verapamil (40 ng/mL) increased significantly both tissue accumulation of [3H] vinblastine and [14C]doxorubicin (to 0.060 +/- 0.024 and 0.034 +/- 0.012 ng/mg protein/h respectively) and serosal transfer (to 0.006 +/- 0.002 and 0.023 +/- 0.001 ng/mg protein/h) (p < 0.05 in all cases). The reproducibility of this in vitro model suggests that the rat everted gut sac is a useful screening tool for studying transport of p-gp substrates and potential p gp modifiers. PMID- 11062738 TI - Induced G2/M arrest and apoptosis in human epidermoid carcinoma cell lines by semisynthetic drug Ukrain. AB - Exposure of ME180 and A431 carcinoma cells to Ukrain (NSC-631570), a novel semisynthetic drug from Chelidonium majus L, results in cell growth inhibition which is concomitant with reversible G2/M cell cycle arrest and apoptosis at doses as low as 7 microM. In contrast, the same drug concentrations were not affective towards normal human keratinocytes. In order to investigate whether cell cycle control mechanisms are effected in response to Ukrain, we analyzed cell cycle distribution and levels of cyclins and cyclin-dependent kinases in drug treated carcinoma cells. We found alterations in levels of mitotic cyclins A and B1, and cyclin-dependent kinases CDK1 and CDK2, after treatment. We also observed an upregulation of CDK inhibitor p27 in both cancer cell lines which may lead to the G2/M cells accumulation. PMID- 11062739 TI - Anticancer effect of 4-[3,5-bis(trimethylsilyl)benzamido] benzoic acid (TAC-101) against A549 non-small cell lung cancer cell line is related to its anti-invasive activity. AB - We examined the effects of TAC-101 on the invasion and metastasis of human non small cell lung cancer (NSCLC) cell lines. TAC-101 showed an ability to inhibit in vitro invasiveness of NSCLC at a non-cytotoxic concentration range of 3-10 microM; such concentration levels were easily achievable following oral administration of therapeutically effective doses. The inhibition of cell invasion at 10 microM of TAC-101 accounted for 58-69% when compared with control cells. Oral administration of TAC-101 (4 mg/kg/day) to mice bearing lung implanted A549 lung cancer resulted in significant life-prolonging effect (T/C: 143%). More pronounced life-prolonging effect was observed in the experimental liver metastasis model of A549, where T/C of 215% was observed following administration at 4 mg/kg/day of TAC-101. However, TAC-101 did not show the direct anti-tumor effect against the established A549 tumor xenografts after subcutaneous implantation. These findings suggest that TAC-101 interferes with cell-to-cell interaction processes leading, for instance, to the inhibition of the invasion of NSCLC cells. Taking into account the pharmacological properties of TAC-101, it is expected that TAC-101 may be a suitable candidate drug for the treatment of lung cancer patients, especially those with a predictable metastasizing potential. PMID- 11062740 TI - Expression patterns of L-plastin isoform in normal and carcinomatous breast tissues. AB - Plastins are members of a family of actin-binding proteins which exhibit a tissue specific expression pattern. L-plastin, which is specifically expressed in hematopoietic cell lineage, has been proposed to be involved in the control of cell adhesion and motility. This protein is also frequently expressed in cell lines derived from mammary solid tumors and therefore might be involved in cancer invasion and metastasis. We have analysed plastin expression in normal and carcinomatous breast tissues in vivo by immunohistochemistry and immunoblotting approaches using specific plastin isoform antibodies. L-plastin was not detected in normal epithelial cells of the mammary gland whereas a staining of myoepithelial cells was observed in 50% of the cases. In breast carcinomas, a significant immunostaining of malignant epithelial cells was observed in 4 of the 29 cases analysed (13.8%). No correlation between L-plastin expression and tumor size, histological grade or lymph node status was observed. In contrast, L plastin was found expressed in 4 of the 11 estrogen and progesterone receptors negative tumors (p = 0.039). The potential role of plastin expression in the tumor process is discussed. PMID- 11062741 TI - Antiapoptotic and antigenotoxic effects of N-acetylcysteine in human cells of endothelial origin. AB - N-Acetylcysteine (NAC) is a drug bearing multiple preventive properties that can inhibit genotoxicity and carcinogenicity. NAC also inhibits invasion and metastasis of malignant cells, as well as tumor take. We recently demonstrated the effects of NAC on Kaposi's sarcoma cells supernatant-induced invasion in vitro and angiogenesis in vivo. Many anticancer agents act through cytotoxicity of rapidly proliferating cells and several antineoplastic drugs induce apoptosis of cancer cells. Since endothelial cells are the target for the inhibition of angiogenesis, we wanted to verify that NAC, while inhibiting tumor vascularization and endothelial cell invasion would not induce endothelial cell apoptosis. We tested the ability of NAC to modulate apoptosis and cytogenetic damage in vitro and to promote differentiation on a reconstituted basement membrane (matrigel) in two endothelial cell lines (EAhy926 and HUVE). Treatment with NAC protected endothelial cells from TGF-beta-induced apoptosis and paraquat induced cytogenetic damage. Therefore, NAC acts as an antiangiogenic agent and, at the same time, appears to prevent apoptosis and oxygen-related genotoxicity in endothelial cells. PMID- 11062742 TI - A novel use for the comet assay: detection of topoisomerase II inhibitors. AB - BACKGROUND: The simple and quick comet assay can quantitatively detect DNA cleavage in cells. This study aimed to determine whether the comet assay could be used to detect topoisomerase (topo) II inhibitors. MATERIALS AND METHODS: HT-29 colon cancer cells were pre-incubated with aclarubicin, a topo II antagonist, then treated with topo II poisons: etoposide (VP-16), teniposide (VM-26), 4' (acridinylamino) methansulfon-m-anisidide (m-AMSA) and adriamycin (doxorubicin). We also tested a topo I poison (camptothecin) and a microtubule depolymerization inhibitor (taxol). RESULTS: Aclarubicin significantly reduced DNA cleavage induced by topo II poisons, but not that induced by camptothecin. In HL-60/MX2 cells (containing no topo II beta and reduced topo II alpha), DNA breakage induced by topo II poisons was lower. Also, aclarubicin antagonized topo I mediated camptothecin-induced DNA cleavage in these resistant cells. CONCLUSIONS: The comet assay can be used to detect topo II poisons in cultured cells. Also, aclarubicin has a dual topo I and topo II antagonism, with "preferential antagonism" of topo II when topo II beta catalytic activity is normally expressed. PMID- 11062743 TI - N-3 polyunsaturated fatty acids accentuate B16 melanoma growth and metastasis through suppression of tumoricidal function of T cells and macrophages. AB - BACKGROUND: The antitumor effects of the n-3 polyunsaturated fatty acids (PUFAs) are still controversial and as yet undefined. MATERIALS AND METHODS: EPA-28, a fish oil enriched with n-3 PUFAs including eicosapentaenoic and docosahexaenoic acids, was administered subcutaneously into C57BL/6 mice before and after subcutaneous inoculation of B16 melanoma cells. The effects of EPA-28 on the antitumor activities of T cells and macrophages were investigated. RESULTS: The treatment of the mice with EPA-28 before and after the tumor inoculation enhanced the growth and metastasis of B16 melanoma and decreased the survival rate of the tumor-bearing mice. The treatment also decreased the number of CD4+ T cells in the spleen and tumor draining lymph nodes on day 14 after the tumor inoculation. Moreover, EPA-28 suppressed the antimelanoma cytolytic activity of T cells and macrophages of the tumor-bearing mice. CONCLUSION: The results suggest that EPA 28 treatment increased both the growth and metastasis of B16 melanoma cells by suppressing the cytolytic function of both T cells and macrophages. PMID- 11062744 TI - Tetracationic porphyrins inhibit angiogenesis induced by human tumor cells in vivo. AB - BACKGROUND: Cationic porphyrin TMPyP4, but not its isomer TMPyP2, inhibits telomerase in tumor cells in vitro and induces chromosome destabilization in vivo. MATERIALS AND METHODS: To examine the effects of these porphyrins on tumor induced angiogenesis, 25-200 micrograms TMPyP4 or TMPyP2 were injected daily for 3 days in mice with intradermally implanted primary human tumor cells. Alternatively, tumor cells were exposed for 90 minutes to 2.5-20 microM porphyrins prior to implantation in mice. RESULTS: Either subcutaneous injections (> or = 50 micrograms/mouse) or preincubation with > or = 5 microM porphyrins significantly inhibited angiogenesis. CONCLUSION: Antiangiogenic activity is apparently unrelated to the ability of the porphyrins to inhibit telomerase. PMID- 11062745 TI - Damage of the kinesin heavy chain gene contributes to the antagonism of cisplatin and paclitaxel. AB - Paclitaxel (TAX) and cisplatin (DDP) rank among the most promising anti neoplastic agents for the treatment of epithelial cancer. Pre-clinical and clinical data show that the cytotoxicity resulting from the combined use of TAX and DDP is governed by the order (TAX prior DDP) > (DDP prior TAX). Since this also occurs with DNA damaging agents other than DDP, damage of a sensitivity gene may be of importance. Recently, a role of kinesin heavy chain (KHC) was uncovered for the cytotoxicity of many natural drug derivatives by use of genetic suppressor elements (Gudkov et al. (1994) Proc. Natl. Acad. Sci. USA, 91, 3744 3748). We first confirmed the dependency on the order of addition for TAX and DDP in human leukemic K562 cells taking apoptotic fraction and cell growth as endpoints. We then inhibited KHC gene expression by antisense oligodeoxy nucleotides or KHC protein function by monoclonal antibody SUK4. Both approaches led to a reduced cytotoxicity of TAX indicating that KHC may confer sensitivity to TAX. Using a PCR-stop assay, we found that DDP indeed caused dose related DNA damage in the KHC gene. There was more DNA damage in the KHC gene than in four other genes (AFP, G6PD, MDR1, TCR-delta). Increasing doses of DDP down-regulated KHC mRNA expression more than of G6PD. There may therefore be a role of KHC for the use of TAX and DDP in combination. PMID- 11062746 TI - Characterization of chemoresistance mechanisms in a series of cisplatin-resistant transitional carcinoma cell lines. AB - We explored the mechanisms of cisplatin resistance in a series of bladder transitional carcinoma cells that are either sensitive or progressively resistant to cisplatin. Resistant lines were raised by chronic exposure of the parental cells to progressively increased concentrations of cisplatin. The cisplatin IC50s of the sensitive and the three resistant cells were 4.3, 25.0, 40.4, and 52.2 microM, respectively. The expressions of glutathione S-transferase pi (GST-pi) and multidrug resistance-associated protein (MRP) were enhanced in a dose response manner as cells acquired progressive cisplatin resistance. Expression of mdr-1 transcript was detected in the three resistant lines but not in the sensitive line. Glutathione contents were increased in resistant cells, yet the trend of increase did not reach statistical significance (p = 0.061). In conclusion, transitional carcinoma cells may gain cisplatin resistance through multiple pathways including up-regulation of GST-pi, MRP and possibly mdr-1. Glutathione contents may play a less significant role in cisplatin chemoresistance. PMID- 11062747 TI - Suppressive effects of Adenophora triphylla extracts on in vitro tumor cell growth and in vivo gastric epithelial proliferation. AB - Adenophora triphylla (AT), an oriental medicinal plant, was extracted using water and several organic solvents and each fraction was assayed for its tumoricidal effects on human Jurkat T cells with 3-(4,5-dimethylthiazolyl)-2,5 diphenyltetrazolium bromide (MTT). The influence on induction of apoptosis and G1 arrest was also examined. The ethyl acetate fraction showed the most pronounced inhibitory effects on proliferation of Jurkat T cells. Apoptosis was induced in line with up-regulation of FasL, tyrosine phosphorylation and c-fos mRNA levels. Arrest in G1 of the cell cycle was observed in A2780 cells with a wild type p53 gene but not HT-29 cells with a mutant p53 gene. Modifying effects of AT on cell turnover and glutathione(GSH) levels in vivo were also investigated in the stomach of rats given 150 mg/kg of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) by gavage and then fed a diet supplemented with 5% or 1% pulverized AT and 0.5% or 0.2% ethylacetate-extracted AT for 42 hours. The 5% AT and both of the ethylacetate fractions caused significant reduction in proliferating cell nuclear antigen (PCNA)-labeling in the glandular stomach epithelium as compared with the value for the MNNG alone group. In addition, the treatments significantly increased the gastric GSH levels. These results suggest that AT could be a chemopreventive agent against gastric cancer. PMID- 11062748 TI - Modulation of the antiproliferative activity of anticancer drugs in hematopoietic tumor cell lines by the poly(ADP-ribose) polymerase inhibitor 6(5H) phenanthridinone. AB - Poly (ADP-ribose) polymerase (PARP) is involved in the cellular responses to genotoxic damage and its inhibition has been proposed as potentiating anticancer drug activity. Here, we evaluated the ability of the PARP inhibitor, 6(5H) phenanthridinone, to modulate the antiproliferative activity of bleomycin, carmustin and doxorubicin in a murine (RDM4) and a human (U937) lymphoma cell lines. 6(5H)-phenanthridinone was shown to suppress PARP activity with the same potency in both cell lines. At 25 microM, this compound potentiated the activity of carmustin in RDM4 but not in U937 cells. In contrast, 6(5H)-phenanthridinone failed to affect the doxorubicin toxicity in murine lymphoma cells, whereas it prevented the cytotoxicity of this drug in the human cell line. Altogether, these findings indicated that 6(5H)-phenanthridinone modulates the cytotoxicity of anticancer agents differently according to the cell type and the drug. Therefore, this PARP inhibitor could be considered as the prototype of a new class of adjuncts in cancer chemotherapy. PMID- 11062749 TI - MEK1 and Erk1/2 kinases as targets for the modulation of radiation responses. AB - BACKGROUND: The activation of mitogenic cascades via Ras, c-Raf-1, MEK-1, Erk1/2 is a hall- mark of oncogenic transformation. The cascade is also anti-apoptotic by modulation of Bcl-2, Bcl-xL and BAD function. The impact of MEK-1 on radiation resistance and the role of drugs targeting MEK-1 for the modulation of resistance is unclear. MATERIALS AND METHODS: Activation of MEK-1 in four carcinoma cell lines was analyzed using Erk1/2 phosphospecific antibodies. MEK-1 was blocked by PD98059 (2-amino-3methoxyflavin) and influence on radiation responses was determined by apoptotic morphology, caspase-3 activation and colony formation assays. RESULTS: MEK-1 kinase was constitutively active and inhibitable by PD98059. The low radiation induced apoptosis rate was not increased by MEK inhibition. PD98059 did not influence cell growth and radiation induced clonogenic cell death. CONCLUSION: Since PD98059 did not alter radiation responses despite blocking MEK-1 kinase, MEK-1 and Erk1/2 are not involved in radiation resistance. Thus PD98059 has no potential in the modulation of radiation responses. PMID- 11062750 TI - Different patterns of inducible nitric oxide synthase gene expression in ovarian carcinoma cell lines. AB - BACKGROUND: Stimulation of inducible nitric oxide synthase (iNOS) and release of nitric oxide by tumor cells may play a critical role in cancer development, either by exhibiting tumoricidal effects or by promoting tumor progression. MATERIALS AND METHODS: In the present study, we investigated four ovarian carcinoma cell lines with respect to their iNOS expressing characteristics. RESULTS: Following incubation with interferon-gamma, tumor necrosis factor-alpha and interleukin-1 beta, a marked stimulation of iNOS gene expression was found in SKOV-6 and OVCAR-3 cells, while only minor iNOS synthesis was detectable in HOC 7. Time-dependent accumulation of nitrite/nitrate in the culture supernatant indicated that the three cell lines were stimulated to produce and release nitric oxide. In contrast, no NO generation was observed in the fourth cell line under investigation, 2774. CONCLUSION: Due to the growth regulatory functions attributed to NO, variations in NO production may be of importance e.g. following interferon-gamma therapy in patients with ovarian cancer. PMID- 11062751 TI - Oxaliplatin exerts potent in vitro cytotoxicity in colorectal and pancreatic cancer cell lines and liver metastases. AB - BACKGROUND: Oxaliplatin displays potent activity in advanced colorectal cancer. The aim of this study was to estimate the potential efficacy of oxaliplatin for hepatic arterial infusion (HAI) chemotherapy. MATERIALS AND METHODS: The anti proliferative effects of oxaliplatin in human HT29 and NMG64/84 colon and COLO 357 MIA PaCa-2 and PMH2/89 pancreatic cancer cell lines and in fresh liver metastases from patients with colorectal and pancreatic cancer were investigated using the human tumor colony forming assay. RESULTS: Oxaliplatin significantly inhibited the colony formation in all cell lines in a concentration- and time dependent manner. All liver tumors displayed a significant concentration dependent inhibition of colony formation after exposure to oxaliplatin for 2 hours. The IC50 of oxaliplatin of 9 of the 10 tumors was < 10 micrograms/ml. CONCLUSION: Oxaliplatin is suitable for HAI therapy phase II studies. Due to the low IC50 values of most tumors we suggest that patients with colorectal or pancreatic liver metastases may benefit from HAI with oxaliplatin. PMID- 11062752 TI - Analysis of the in vitro inhibition of mammary adenocarcinoma cell adhesion by sulphated polysaccharides. AB - Evidence is mounting that changes in the ability of cancer cells to adhere to extracellular matrices (ECM) play a decisive role in metastasis spread. We have investigated the effect of different sulphated polysaccharides on the adhesion of MCF7 and MDA-MB231 adenocarcinoma breast cells to different substrata: a reconstituted basement membrane (Matrigel) and various adhesion-mediating proteins (fibronectin, laminin, type IV collagen). Most of them inhibited cell adhesion and the most active component is a galactose rich units polysaccharide, carrageenan iota. Taken together, the results suggest that this inhibitory activity depends on the charge density related to sulphate groups, the molecular weight and also the carbohydrate structure. These products very likely unstabilize the interaction between the glucosaminoglycan portion of proteoglycans and the ECM proteins and then block the ability of these adhesive proteins to bind to cells. PMID- 11062753 TI - An improved system for quantifying AgNOR and PCNA in canine tumors. AB - BACKGROUND: Quantifying silver stained nucleolar organizer regions (AgNORs) and proliferation cell nuclear antigens (PCNA) are useful techniques to measure proliferative activity of tumor cells; however, the nonspecific deposition of stains and overlappings of AgNOR and PCNA counts between grades of tumors hamper their applications. MATERIALS AND METHODS: Fifty-two surgical specimens from dogs, including mast cell tumors, perianal gland tumors and hyperplasias, fibromas, fibrosarcomas, and normal tissues were studied. The 3 microns dewaxed sections of formalin-fixed tissues were stained to detect AgNORs by a modified inverted incubation technique in a newly developed silver staining device. Data were collected and analyzed using a high-resolution digital microscope camera and image analysis software. Sequential sections were also stained for PCNA using an immunohistochemical method. RESULTS: The improved system for quantifying AgNOR provided more accurate and non-overlapping mean AgNOR counts, which enable us to distinguish benign states from malignant changes. The mean AgNOR cut-off points that discriminated grade II or III mast cell tumors from grade I, perianal gland carcinomas from adenomas (or hyperplasia), fibrosarcomas from non-fibrosarcoma tissues, were 6.0, 14.1, 9.4, and 8.8 respectively. The mean AgNOR areas, relative AgNOR areas, and PCNA positive rates of some malignant and non-malignant tissues (benign tumor and normal tissues) were significantly different (P < 0.05). CONCLUSIONS: This improved system is a sensitive and rather precise method for quantifying the AgNOR and PCNA. It provides a valuable objective measurement for differentiating benign and malignant tumors. PMID- 11062754 TI - Immunocytochemical detection of the homeobox B3, B4, and C6 gene products in breast carcinomas. AB - Breast cancer (BC) represents the most frequent neoplasm in women with a risk of incidence between 10% and 12%. The detection of tumor associated and oncofetal antigen re-expression in a variety of neoplastically transformed cell types has aided in the more precise diagnosis and prognostication of human cancers. The homeobox (HOX) genes encode proteins which contain a 61 amino acid DNA-binding homeodomain and are involved in the transcriptional regulation of other genes during normal onto- and histogenesis. The class I HOX genes are organized in four clusters on different chromosomes in humans, with a high conservation in the order of the genes within each of these clusters. Re-expression of HOX gene products has been reported in a wide variety of neoplastically transformed cells and it seems quite likely that the HOX genes represent yet another class of oncofetal antigens involved in both normal development and carcinogenesis, as well as tumor progression. The expression pattern of three HOX gene products (HOX B3, -B4, and -C6) was examined immunocytochemically in 11 human breast carcinoma (BC) tissues. In all observed BC cases, HOX-C6 was present in over 90% of the neoplastically transformed cells (+4) demonstrating a high grade (A and B) staining intensity. The same expression pattern was defined for the other two observed proteins (HOX-B3 and -B4; over 90% or +4 and a high grade staining intensity or A and B). Current treatment of BC encompasses the three "classic" modalities of therapy: surgical resection, radiotherapy, and chemotherapy. Although advances have been made, we still face great difficulties in the treatment of this deadly human neoplasm. Therefore, we are always seeking novel tumor associated antigens (TAAs), including oncofetal antigens, to use as molecular targets in cancer cell directed fourth modality immunotherapy. PMID- 11062755 TI - Matrix metalloproteinase expression in childhood astrocytomas. AB - Structural changes in the extracellular matrix (ECM) are necessary for cell migration during tissue remodeling and local neoplastic cell invasion. The matrix metalloproteinases (MMPs) and their inhibitors have been shown to be critical modulators of ECM composition and are thus, crucial in tumor cell invasion and metastasis. The immunocytochemical profile of MMP-2, -3, -9, -10, and -13 expression was observed in 24 primary human childhood astrocytomas (ASTRs) employing an indirect alkaline phosphatase conjugated antigen detection technique. Evaluation of the results was based on (a) the percent of neoplastically transformed cells that reacted positively and (b) a measure of staining intensity [graded from A (highest) to D (negative)]. The two forms of stromelysin, MMP-3 and -10, share 82% sequence homology, but exhibit differences in cellular synthesis and inducibility by cytokines and growth factors in vitro. Strong overall expression of MMP-3 and -10 was found in ASTRs, especially in the ECM adjacent to blood vessels. Positive immunoreactivity could be seen for these two MMPs in the ECM surrounding over 90% of the neoplastically transformed cells (+4) and the staining intensity was also the strongest possible (A,B). No immunoreactivity was detected using antibodies directed against MMP-2, -9, and 13. Based on these results, MMP-3 and -10 are implicated in the pathogenesis of pediatric ASTRs. Further characterization of the expression and utilization of MMPs and their inhibitors in the progression of ASTRs may establish differential regulation and utilization of the various MMPs during the progression of glial tumors, from low-grade pilocytic ASTR to high-grade glioblastoma multiforme. PMID- 11062756 TI - Induction of tumor-specific antitumor immunity after chemotherapy with cisplatin in mice bearing MOPC-104E plasmacytoma by modulation of MHC expression on tumor surface. AB - Chemotherapy sometimes results in induction of specific antitumor immunity. We investigated the mechanisms responsible for the induction of antitumor immunity in mice bearing MOPC-104E plasmacytoma after chemotherapy with cisplatin (CDDP), especially the effects of CDDP on the expression of MHC on the tumor surface. BALB/c mice were subcutaneously (s.c.) inoculated with MOPC-104E cells on day 0, then intravenously (i.v.) treated with CDDP at 3.6 mg/kg on day 7. The tumors disappeared completely on day 35 and the mice rejected a second challenge with MOPC-104E, but did not reject syngeneic Meth-A fibrosarcoma. The tumors did not regress, however, when MOPC-104E was s.c. transplanted in nude mice, or when anti T-cell monoclonal antibodies were i.v. injected into BALB/c mice before CDDP treatment on day 6. To determine which of the mice or tumor were affected by CDDP, BALB/c mice were inoculated with CDDP-treated (12.5 micrograms/ml for 3 hours in vitro) MOPC-104E cells on day 0, 7 and 14. The potential to reject MOPC 104E was lower in mice immunized with ethanol-treated MOPC-104E cells than in those immunized with CDDP-treated cells. CDDP-treated or -untreated MOPC-104E cells ultrasonicated and fractionated into soluble and insoluble fractions by centrifuging, also induced antitumor immunity. Flow cytometry demonstrated that the expression of MHC-class-I antigens H-2Dd and H-2Kd was enhanced after CDDP treatment, but that of class-II antigens I-Ad and I-Ed was not, suggesting that CDDP induced tumor-specific antitumor immunity by enhancing the expression of MHC class-I antigens. PMID- 11062757 TI - Expression of p21 in non small cell lung cancer relationship with PCNA. AB - The first cyclin dependent kinase inhibitor to be discovered was the p21 cdk interacting protein (a.k.a., WAF1, Cip1, CAP20, Sdi1, mda6). p21 expression may or may not be dependent on p53. This pathway also inhibits DNA replication by merit of p21's interaction with PCNA, but it has also been shown that this same inhibitory interaction with p21 does not affect PCNA DNA repair abilities. We assessed the immunohistochemical expression of p21 protein in 60 curative surgical resected non small cell lung cancers relating it to the expression of PCNA to clarify the contribution of the p21/PCNA pathway to the development of NSCLC. We did not find any relationship between PCNA and p21 expression. This last result may indicate that the mechanism by which PCNA controls the DNA repair is the most important activity of this protein during lung cancer progression and development, compared to its contribution to cell proliferation. In fact, this last event is strongly counteracted by p21 expression, which in this last case works as an inhibitor of PCNA expression. In conclusion this study highlighted the important role of the p21/PCNA pathway in lung carcinogenesis, pointing out the contribution of PCNA to the response to lung aggression and not only it's role as a proliferation index. Therefore, these results offer a background to further study to evaluate potential novel therapeutic approaches to lung cancer treatment. PMID- 11062758 TI - DNA but not topoisomerases is a target for a cytotoxic benzo[5,6]cyclohepta[b]indol-6-one derivative. AB - The interaction of a newly designed benzocycloheptaindol-6-one derivative with DNA has been investigated by complementary spectroscopic techniques including absorption, circular and linear dichroism. Footprinting measurements were performed to delineate the sequence-selectivity of the drug-DNA interaction and a plasmid relaxation assay was used to study the effects of the drug on human DNA topoisomerases I and II. The results clearly indicated that the test compound behaves as a typical DNA intercalating agent but does not stimulate DNA cleavage by topoisomerases. At the cellular level, the cytometry measurements showed that the drug provoked a marked accumulation of HL60 human leukemia cells in the G2/M phase of the cell cycle. DNA is thus identified as a valid target for this new series of drugs particularly toxic to human (HL60) and murine (P388) leukemia cells. PMID- 11062759 TI - Relationship between DNA-reactivity and cytostatic effect of two novel bile acid platinum derivatives, Bamet-UD2 and Bamet-D3. AB - BACKGROUND AND AIMS: Several platinum(II)-bile acid derivatives, named Bamets, have been previously synthesized. Their ability to interact with DNA, their cytostatic activity and their liver organotropic properties have been characterized. Two new compounds of this family, with particular structural properties, have been developed. Bamet-UD2 was formed by two ursodeoxycholic acid moieties bound by the carboxylate groups to cisplatin. In contrast, in Bamet-D3, glycine and a polyamine were used as tandem spacer elements to separate a cholic acid moiety from the platinum(II) atom. The aim of this work was to evaluate how these changes affect the ability of these compounds to interact with DNA and reduce tumour cell growth. MATERIALS AND METHODS: Drug reactivity with DNA was determined by changes in the electrophoretic mobility of the pUC18 plasmid test and by the ethidium bromide (EthBr) displacement assay. Cytostatic activity was measured against two mouse-derived cell lines from lymphocytic leukemia (L1210) and sarcoma (S-180-II). RESULTS: Bamet-UD2, and more markedly Bamet-D3, induced changes in the electrophoretic mobility of pUC18, suggesting the formation of DNA drug interactions. Bamet-UD2 displaced EthBr from its binding to DNA. This effect was stronger in the case of Bamet-D3. Scatchard plots revealed that pre incubation with both Bamet-UD2 and Bamet-D3 decreased the number of DNA sites available and their ability to bind EthBr. In spite of the enhanced DNA reactivity of Bamet-D3, its ability to reduce tumour cell growth was much weaker than that of Bamet-UD2, which was seen to exert a very strong cytostatic effect. CONCLUSION: Although the distance between the platinum atom and the bile acid moiety affects the in vitro Bamet reactivity with DNA, other factors determine the overall cytostatic activity of these compounds. PMID- 11062760 TI - Effects of citrus phytochemicals on liver and lung cytochrome P450 activity and on the in vitro metabolism of the tobacco-specific nitrosamine NNK. AB - NNK is a potent environmental carcinogen to which both smokers and nonsmokers are exposed. The response to NNK may be affected by factors including nutrition. We investigated the effects of five citrus phytochemicals on the in vitro metabolism of the tobacco-specific nitrosamine NNK and on the dealkylation of methoxyresorufin (MROD) and pentoxyresorufin (PROD) in liver and lung microsomes of the Syrian golden hamster. In the NNK metabolism experiments in vitro incubations contained 3 microCi [5-H3] NNK, 0.5 mg microsomal protein and 0.5 mumole of the citrus phytochemical diosmin, naringin, naringenin, quercetin or rutin. In the dealkylation studies incubations contained 0.5 microM methoxyresorufin or pentoxyresorufin, 0.5 mg microsomal protein and 0.5 mumole of citrus phytochemical. The major NNK metabolism pathway in hamster liver microsomes was NNK-reduction while in lung microsomes it was alpha-hydroxylation. The alpha-hydroxylation pathway produces metabolic products that methylate and pyridyloxobutylate DNA. Naringenin, a metabolite of naringin, and quercetin were the most potent inhibitors of alpha-hydroxylation of NNK in both liver and lung microsomes. This inhibition correlated with a potent inhibition of MROD and PROD activity in liver but not in lung microsomes. The metabolic activation of NNK is associated with cytochrome P450 isoforms 1A1, 1A2, 2B1, 2D6 and 2E1. Our results suggest that naringenin and quercetin from citrus fruits inhibit the activity of cytochrome P450 (CYP) isoforms that activate NNK and may afford protection against NNK-induced carcinogenesis. PMID- 11062761 TI - Modulation of transferrin synthesis, transferrin receptor expression, iNOS expression and NO production in mouse macrophages by cytokines, either alone or in combination. AB - Iron, an essential element for all living organisms, is central importance in a number of crucial metabolic pathways, including the regulation of immune function. Iron delivery to cells is accomplished by the complexing of iron to transferrin (Tf), a monomeric iron-binding protein in the plasma, followed by specific binding of Tf to cell-surface receptors, endocytosis of the receptor ligand complexes and ultimately, release of iron from endosomal vesicles to the cytoplasm. The purpose of this study was to evaluate the effect of cytokines, alone and in combination, on the factors that can affect the iron delivery in thioglycollate-elicited macrophages. In this study, IFN gamma induced a marked increase in Tf synthesis by macrophages, while IL-1, IL-6 and TNF alpha produced a more modest increase. Combinations of these cytokines were shown to be less effective in promoting macrophage Tf synthesis than the cytokines by themselves. IFN gamma alone and in combination with other cytokines was effective in inducing nitrite (NO) production and inducible nitric oxide synthetase (iNOS) expression in macrophages, while IL-1, TNF alpha and IL-6 individually, as well as in various combinations, were not. While all tested cytokines individually and in combination inhibited the expression of the transferrin receptor (TfR) on macrophages, IFN gamma alone and in combination with other cytokines most strongly repressed the TfR expression. TfR localization in macrophages after IFN gamma stimulation showed that TfR fluorescence was most intense in the perinuclear region after 6 hours and scattered diffusely throughout the cytoplasm after 24 hours. This data suggests that IFN gamma may enhance iron uptake during the early phase of macrophage activation, and in later phases, down-regulate TfR expression by inducing NO, thus contributing to intracellular oxidative stress reduction. PMID- 11062762 TI - Tissue dosimetry of liposome-radionuclide complexes for internal radiotherapy: toward liposome-targeted therapeutic radiopharmaceuticals. AB - BACKGROUND: Quantitative examination of the important physical parameters, such as the tumor absorbed dose and the tumor-to-normal-tissue (T-NT) absorbed dose ratios, for effective use of radionuclide-liposome conjugates m internal radiotherapy was carried out. METHODS: The Medical Internal Radiation Dose (MIRD) formalism was used to develop a set of dosimetric equations. Pharmacokinetic functions used as input information to the dosimetric model were derived from experimental time-biodistribution data. Multilamellar (MLV), small unilamellar (SUV) and sterically stabilized (GM1- and PEG-coated) liposomes were examined in combination with the very promising particle emitting radionuclides: 67Cu, 188Re and 211At. For comparative purposes, the widely used: 90Y and 131I were also included in the study. For all radionuclide-liposome combinations, the mean absorbed dose per amount of radioactivity administered was obtained: (i) for two different types of human xenografts located in the muscle and liver tissue, and (ii) for normal liver, spleen, kidneys, and total body. RESULTS: Regardless of radionuclide, the poorest values were obtained for the MLV liposomes. Due to more rapid uptake of sterically stabilized (GM,-coated) liposomes to the muscle tumor tissue as compared to SUVs, 211At and 188Re deliver higher tumor doses when combined with the former, while 67Cu, 90Y and 131I are more effective with SUVs. The most promising results were obtained for the [211At-GM1] complex in the liver tumor. CONCLUSION: The importance of liposome size and steric barrier when designing effective radionuclide-carrier systems was revealed, but most importantly the optimal matching between the radionuclide half-life and the time of maximum liposome accumulation ratio between the tumor and normal tissue. PMID- 11062763 TI - Harvesting the human genome: the Israeli perspective. AB - The post-genome era is at our door, and soon the complete human genome sequence will be available for the next set of goals. Israel is well equipped and skilled to join the worldwide harvest of the human genome, but additional massive government investment is required. This will affect various domains of activity, including the fields of diagnostics and therapeutics. The technologies and know how described above constitute the basis for future human genome applications in Israel. PMID- 11062764 TI - Screening for genetic disorders among Jews: how should the Tay-Sachs screening program be continued? AB - The screening program in Israel for Tay-Sachs disease has proven very successful, giving Jewish couples a choice not to have affected children. The technology of carrier detection is now possible in several other severe genetic diseases that are relatively frequent among Jews. Due to the current confusion, a policy is needed to determine how the TSD screening program should be continued in the Israeli Jewish population. We propose that such a screening program include only mutations agreed by consensus as causing a disease severe enough to warrant the possibility of therapeutic abortion. We also propose that general screening include only mutations that are relatively frequent, taking into account the carrier frequencies in the Israeli Jewish population. PMID- 11062765 TI - Serum total cholesterol and cardiovascular mortality in Israeli males: the CORDIS Study. Cardiovascular Occupational Risk Factor Determination in Israeli Industry. AB - BACKGROUND: The degree to which serum total cholesterol predicts cardiovascular disease is uncertain. While most authors have placed TC among the most powerful risk indicators of CVD, some have claimed that it predicted CVD in women only, or even not at all. OBJECTIVE: To determine the predictive value of serum total cholesterol relative to diabetes, smoking, systolic blood pressure and body mass index (kg/m2), for cardiovascular disease mortality in 3,461 occupationally active Israeli males. METHODS: A prospective follow-up was carried out for the years 1987-1998 to determine the effect of age, smoking habits, a history of diabetes, SBP, BMI and TC, at entry, on CVD mortality. RESULTS: There were 84 CVD deaths during a total of 37,174 person-years follow up. The hazard ratios (95% confidence intervals) for CVD mortality with respect to variables at entry were: diabetes 5.2 (2.1-13.2), age 2.2 (1.7-2.9), smoking 1.3 (1.0-1.8), SBP 1.4 (1.1 2.0), TC 1.5 (1.0-2.1) and BMI 1.2 (0.7-2.2). Among non-obese, non-diabetic, normotensive subjects the hazard ratio of TC adjusted for age and smoking was 1.16 (1.09-1.22) per 10 mg/dl. In the remaining subjects it was 1.04 (0.98-1.12) only. There was a significant interaction between TC and diabetes, hypertension or obesity (P = 0.003). CONCLUSIONS: In this population of Israeli males we found an interaction between TC and other risk indicators for CVD. Confirmation is required for the unexpected finding that the predictive value of TC for CVD mortality among non-diabetic, non-obese and normotensive subjects exceeded that among subjects with either of these risk factors. PMID- 11062766 TI - Outcome of "out of hospital" cardiopulmonary arrest in children admitted to the emergency room. AB - BACKGROUND: The outcome of cardiopulmonary arrest in children is poor, with many survivors suffering from severe neurological defects. There are few data on the survival rate following cardiopulmonary arrest in children who arrived at the emergency room without a palpable pulse. OBJECTIVE: To determine the survival rate and epidemiology of cardiopulmonary arrest in children who arrived without a palpable pulse at a pediatric ER in southern Israel. METHODS: We retrospectively reviewed the medical records of all patients with cardiopulmonary arrest who arrived at the ER of the Soroka University Medical Center during the period January 1995 to June 1997. RESULTS: The study group included 35 patients. Resuscitation efforts were attempted on 20, but the remaining 15 showed signs of death and were not resuscitated. None of the patients survived, although one patient survived the resuscitation but succumbed a few hours later. The statistics show that more cardiopulmonary arrests occurred among Bedouins than among Jews (32 vs. 3, P < 0.0001). CONCLUSIONS: The probability of survival from cardiopulmonary arrest in children who arrive at the emergency room without palpable pulse is extremely low. Bedouin children have a much higher risk of suffering from out-of-hospital cardiopulmonary arrest than Jewish children. PMID- 11062767 TI - Screening intention and practice among first-degree relatives of colorectal cancer patients in southern Israel. AB - BACKGROUND: First-degree relatives of colorectal cancer patients are the largest groups of individuals at increased risk for colorectal cancer. OBJECTIVE: To assess the knowledge, attitudes and behavior to disease prevention and colorectal cancer screening among first-degree relatives of colon cancer patients. METHODS: A descriptive, point-prevalence epidemiological study was conducted among 215 first-degree relatives of survivors of colorectal cancer in the southern (Negev) region of Israel. Variables included perceived health status, knowledge about cancer screening, compliance rates with colorectal cancer screening, and interest in participation in early detection programs in the future. RESULTS: The mean age of the respondents was 47.9 +/- 11.2 years, and 54% were males. Only 58 (27%) remembered having been encouraged to undergo an early detection test. In the previous year only 15% underwent fecal occult blood tests, while 9% had a barium enema and 14% an endoscopic examination of the colon by sigmoidoscopy or colonoscopy. A total of 49% of the asymptomatic respondents were unaware of recommendations for screening, and only 38.3% expressed any interest in participating in early detection programs in the future. Only 19% of respondents over the age of 50 and 8% of respondents over age 60 were interested in participating compared with 49% under the age of 50 (P < 0.0001). CONCLUSION: A minority of first-degree relatives of colorectal cancer patients reported having been counseled to undergo screening, although most had seen their family physician in the previous 3 years. Primary care physicians should be more active in informing at-risk patients and encouraging them to undergo periodic screening. PMID- 11062768 TI - Gynecological aspects of female familial dysautonomia. AB - BACKGROUND: Familial dysautonomia is a genetic disease in which there is a defect in the autonomic and sensory nervous systems. These systems have a major role in the reproductive system. OBJECTIVE: To study the inter-relationship of autonomic and sensory dysfunction and gynecological function. METHODS: The gynecological histories of 48 women with familial dysautonomia were analyzed retrospectively. Their mean age was 22.25 years (range 12-47). Thirty-three women (65%) were available for further questioning and investigation of hormonal status. RESULTS: Menarche had occurred in 32 of the 48 (66.7%). Their average age of menarche was significantly delayed as compared to their unaffected mothers (15.5 vs. 13.6 years respectively, P = 0.002). The most prominent finding was the very high prevalence, 81.2%, of premenstrual symptoms. Seven of 26 had premenstrual syndrome symptoms of dysautonomic crisis. Blood sex hormone levels were normal in 27 of the 33 patients studied. None reached natural menopause. One patient had adenomyosis, and another, dysgerminoma. Three patients became pregnant and delivered healthy infants. CONCLUSION: Menarche is delayed in women with FD, and the physiological monthly hormonal fluctuations may disturb autonomic homeostasis sufficiently to precipitate dysautonomic crisis. PMID- 11062769 TI - Background diseases in 671 patients with moderate to severe pulmonary hypertension. AB - BACKGROUND: Data regarding the epidemiology of secondary pulmonary hypertension are scanty. OBJECTIVES: To describe the spectrum and relative incidence of background diseases in patients with significant secondary PHT. METHODS: We identified 671 patients with systolic pulmonary artery pressure of 45 mm Hg or more from the database of the echocardiographic laboratory. Their background diseases were recorded and classified into three subgroups: cardiac, pulmonary and pulmonary vascular disease without pulmonary parenchymal disease. Age at the first echocardiographic study, gender and systolic PAP values were recorded. Data between the three subgroups were compared. RESULTS: The mean age of the patients was 65 +/- 15 years, mean systolic PAP 61 +/- 14 mm Hg and female:male ratio 1.21:1. At the time of diagnosis 85% of the patients were older than 50. PHT was secondary to cardiac disease in 579 patients (86.3%), to PVD without PPD in 54 patients (8%) and to PPD in only 38 patients (5.7%). Mean age and mean systolic PAP did not differ significantly among the three subgroups. There was a significantly higher female:male ratio in patients with PVD without PPD compared with cardiac or pulmonary diseases (1.7:1 vs. 1.2:1 and 1.7 vs. 0.8:1 respectively, P < 0.05). CONCLUSIONS: The majority of patients with significant PHT are elderly with heart disease. PVD without PPD and chronic PPD are a relatively uncommon cause of significant PHT. Since the diagnosis of PHT is of clinical significance and sometimes merits different therapeutic interventions, we recommend screening by Doppler echocardiography for patients with high risk background diseases. PMID- 11062770 TI - The future of genetics: where are we going in the next forty years? PMID- 11062771 TI - Cardiopulmonary resuscitation in children admitted to the emergency room without a pulse following out-of-hospital cardiac arrest: too little too late? PMID- 11062772 TI - Non-steroidal anti-inflammatory drugs and selective apoptotic anti-neoplastic drugs in the prevention of colorectal cancer: the role of super aspirins. AB - There is increasing evidence to suggest that aspirin and other non-steroidal anti inflammatory drugs reduce the risk of colorectal cancer. This observation is supported by animal studies that show fewer tumors per animal and fewer animals with tumors after administration of several different NSAIDs. Intervention data in familial adenomatous polyposis have established that the effect is exerted on the process of human colonic adenoma formation. Supportive evidence in sporadic colorectal neoplasia, derived from 22 of 24 studies (both case-control and cohort), found a reduced risk in men and women for cancers of the colon and the rectum and for both aspirin and the other NSAIDs. Earlier detection of lesions as a result of drug-induced bleeding does not seem to account for these findings. Although the molecular mechanism responsible for the chemopreventive action of this class of drugs is not yet completely understood, the protection may affect several pathways including both cell cycle arrest and induction of apoptosis. In the third millennium the question is not if but how. Based on the consistency of epidemiological, clinical and experimental data, the association between regular long-term aspirin or NSAIDs intake and a decreased death rate from colorectal cancer is sound and there is no need for further placebo trials. At the same time, despite this consistency there is no clear data on the dose, duration or frequency of use for cancer-preventive activity. PMID- 11062773 TI - Aging and the human immune system. PMID- 11062774 TI - Dextromethorphan in chronic pain: a disappointing update. PMID- 11062775 TI - Suction purpura. PMID- 11062776 TI - Sarcoidosis with cholangitis component in a patient with autoimmune hemolytic anemia. PMID- 11062777 TI - Caution when using oral contraceptive pills with Orlistat. PMID- 11062778 TI - Meningoencephalitis caused by Rickettsia typhi. PMID- 11062779 TI - Fibrosing omental panniculitis and polyserositis associated with long-term treatment by paroxetine. PMID- 11062780 TI - Right ventricular rupture following postoperative mediastinitis: a call for caution. PMID- 11062781 TI - Small bowel perforation within an inguinal hernia due to a foreign body. PMID- 11062782 TI - Follicular carcinoma of the thyroid presenting as back pain and paravertebral mass. PMID- 11062783 TI - Comprehensive lipid analysis: a powerful metanomic tool for predictive and diagnostic medicine. AB - The power and accuracy of predictive diagnostics stand to improve dramatically as a result of lipid metanomics. The high definition of data obtained with this approach allows multiple rather than single metabolites to be used in markers for a group. Since as many as 40 fatty acids are quantified from each lipid class, and up to 15 lipid classes can be quantified easily, more than 600 individual lipid metabolites can be measured routinely for each sample. Because these analyses are comprehensive, only the most appropriate and unique metabolites are selected for their predictive value. Thus, comprehensive lipid analysis promises to greatly improve predictive diagnostics for phenotypes that directly or peripherally involve lipids. A broader and possibly more exciting aspect of this technology is the generation of metabolic profiles that are not simply markers for disease, but metabolic maps that can be used to identify specific genes or activities that cause or influence the disease state. Metanomics is, in essence, functional genomics from metabolite analysis. By defining the metabolic basis for phenotype, researchers and clinicians will have an extraordinary opportunity to understand and treat disease. Much in the same way that gene chips allow researchers to observe the complex expression response to a stimulus, metanomics will enable researchers to observe the complex metabolic interplay responsible for defining phenotype. By extending this approach beyond the observation of individual dysregulations, medicine will begin to profile not single diseases, but health. As health is the proper balance of all vital metabolic pathways, comprehensive or metanomic analysis lends itself very well to identifying the metabolite distributions necessary for optimum health. Comprehensive and quantitative analysis of lipids would provide this degree of diagnostic power to researchers and clinicians interested in mining metabolic profiles for biological meaning. PMID- 11062784 TI - Neurological aspects of the David-Goliath battle: restriction in the giant's visual field. PMID- 11062785 TI - Abdominal aortic aneurysm: endovascular repair. PMID- 11062786 TI - Lower-limb amputation and diabetes: the key is prevention. PMID- 11062787 TI - Reinventing ourselves. PMID- 11062788 TI - Screening for abdominal aortic aneurysm: lessons from a population-based study. AB - OBJECTIVES: To test the acceptability of screening and to identify modifiable risk factors for abdominal aortic aneurysm (AAA) in men. DESIGN: A trial of ultrasound screening for AAA in a population-based random sample of men aged 65 83 years, and a cross-sectional case-control comparison of men in the same sample. PARTICIPANTS: 12,203 men who had an ultrasound examination of their abdominal aorta, and completed a questionnaire covering demographic, behavioural and medical factors. MAIN OUTCOME MEASURES: Prevalence of AAA, and independent associations of AAA with demographic, medical and lifestyle factors. RESULTS: Invitations to screening produced a corrected response of 70.5%. The prevalence of AAAs (> 30 mm) rose from 4.8% in men aged 65-69 years to 10.8% in those aged 80-83 years. The overall prevalence of large (> 50 mm) aneurysms was 0.69%. In a multivariate logistic model Mediterranean-born men had a 40% lower risk of AAA (> 30 mm) compared with men born in Australia (odds ratio [OR], 0.6; 95% CI, 0.4 0.8), while ex-smokers had a significantly increased risk of AAA (OR, 2.3; 95% CI, 1.9-2.8), and current smokers had even higher risks. AAA was significantly associated with established coronary and peripheral arterial disease and a waist:hip ratio greater than 0.9; men who regularly undertook vigorous exercise had a lower risk (OR, 0.8; 95% CI, 0.7-1.0). CONCLUSION: Ultrasound screening for AAA is acceptable to men in the likely target population. AAA shares some but not all of the risk factors for occlusive vascular disease, but the scope for primary prevention of AAA in later life is limited. PMID- 11062789 TI - Diabetes-related lower-limb amputations in Australia. AB - OBJECTIVE: To identify the prevalence of diabetes-related lower-limb amputations and its regional variations in Australia. DESIGN AND SETTING: Cross-sectional analysis of a hospital morbidity dataset in Australia. METHODS: Analysis of the National Hospital Morbidity Database of all hospital separations for the ICD codes 84.10-84.19 (lower-limb amputations) and 250.0-250.9 (diabetes and its complications) for the financial years 1995-96 to 1997-98. MAIN OUTCOME MEASURE: Number of lower-limb amputations in people with diabetes mellitus in Australia, and in each State and Territory. RESULTS: 7887 diabetes-related lower-limb amputations were reported during the study period, with a mean +/- SD of 2629 +/- 47 per year. The prevalence in Australia was 13.97 per 100,000 total population, and varied from 11.34 per 100,000 in the Australian Capital Territory to 20.68 per 100,000 in South Australia. CONCLUSION: Diabetes-related lower-limb amputation poses a substantial personal and public health cost in Australia. PMID- 11062790 TI - Potential roles for quantitative ultrasound in the management of osteoporosis. AB - OBJECTIVE: To assess the validity of four models for the role of quantitative ultrasound (QUS) in the management of osteoporosis. DESIGN: Cross-sectional survey and review of literature. SETTINGS: Nuclear medicine departments of three teaching hospitals in Sydney. SUBJECTS: 1000 women aged 22 to 88 years (mean, 59 years) referred for assessment of osteoporotic fracture risk. MAIN OUTCOME MEASURES: BMD categories as defined by dual-energy x-ray absorptiometry (DEXA) of the lumbar spine and proximal femur, and QUS category as defined by calcaneal ultrasound stiffness; prevalence of DEXA-defined osteoporosis in the different QUS categories. RESULTS: In women with QUS Achilles stiffness < or = 70 the prevalence of axial osteoporosis was 51%, whereas in the group with stiffness > 70 the prevalence of axial osteoporosis was 8%. In women 65 years and over the corresponding values were 59% and 17%. CONCLUSIONS: Of the four possible models for QUS, the use of QUS for the estimation of BMD, or in a "standalone" model, can not be recommended at the current time. The model of QUS as a "prescreening" modality may be acceptable assuming adequate education of clinicians and patients of its limitations, particularly the risk of false negatives. The model of QUS as one factor in a composite risk factor assessment of patients is promising but more data are required. PMID- 11062791 TI - Sertraline treatment of interferon-alfa-induced depressive disorder. AB - Interferon alfa therapy for chronic hepatitis C infection is commonly associated with neuropsychiatric symptoms, including depression. These side effects may necessitate reduction or even cessation of interferon alfa, but there is little information regarding the management of this important problem. We report 10 cases of interferon-alfa-induced depressive disorder treated with the selective serotonin reuptake inhibitor sertraline. All patients obtained rapid symptom relief without the need for reduction or cessation of interferon alfa. PMID- 11062792 TI - Domestic violence in Australia: definition, prevalence and nature of presentation in clinical practice. AB - Domestic violence is a complex pattern of behaviours that may include, in addition to physical acts of violence, sexual abuse and emotional abuse. Women experience domestic violence at far greater rates than men do, and women and children often live in fear as a result of the abuse that is used by men to maintain control over their partners. Domestic violence is a major public health problem and is very common in women attending clinical practice. Women present most commonly with a range of chronic symptoms to unsuspecting general practitioners, emergency department doctors or medical specialists. Women who have experienced partner abuse want to be asked about it and are more likely to disclose if asked in an empathic, non-judgemental way. Doctors can make a difference. PMID- 11062793 TI - The lower limb in people with diabetes. Position statement of the Australian Diabetes Society. AB - Diabetic lower-limb problems result in significant social, medical and economic consequences and are the most common cause of hospitalisation for people with diabetes. In people with diabetes, amputations are 15 times more common than in people without diabetes, and 50% of all amputations occur in people with diabetes. Peripheral neuropathy, vascular disease, infection and deformity of the feet are the major predisposing factors leading to ulceration or amputation. All people with diabetes should receive basic footcare education, and regular foot examinations. The risk for the development of ulceration can be assessed by basic clinical examination of the foot. Management strategies depend on the risk category, and range from basic education and annual review to specialist care by a multidisciplinary team. PMID- 11062794 TI - Clinical practice guidelines and the computer on your desk. AB - The majority of Australia's general practitioners are now believed to be using a computer to support clinical practice. This technology has the potential to deliver evidence-based clinical practice guidelines to the GP during consultations. If clinical practice guidelines are to be incorporated into electronic medical record systems, to assist clinical decision making at the point of care, guideline development will need to be significantly revised. Computerised clinical decision support at the time of consultation may become a major method for continuing medical education for Australian GPs. PMID- 11062795 TI - The intersection of health informatics and evidence-based medicine: computer based systems to assist clinicians. AB - Evidence-based medicine and health informatics may assist clinicians contend with the current barriers to the integration of evidence-based "best practice" into typical healthcare settings. Computer-based decision support systems can improve the process of care, but whether they improve patient outcomes in a cost effective manner is uncertain. PMID- 11062796 TI - Screening gamete donors for cystic fibrosis status. AB - Gamete donors are currently not tested for cystic fibrosis, even though carriers have a very high risk of producing children with the disease. We recommend that gamete donors be routinely tested for cystic fibrosis. Similar arguments exist for antenatal screening for cystic fibrosis. PMID- 11062797 TI - Disruptive doctors. Unprofessional interpersonal behaviour in doctors. PMID- 11062798 TI - Consumption of dietary supplements and energy drinks by schoolchildren. PMID- 11062799 TI - Headache and face pain. PMID- 11062800 TI - Active and passive cigarette smoking and breast cancer: is a real risk emerging? PMID- 11062801 TI - Retreat of the pneumococcus? PMID- 11062802 TI - Epidemiology of invasive pneumococcal disease in urban New South Wales, 1997 1999. AB - OBJECTIVES: To describe the serotypes, incidence and morbidity of invasive pneumococcal disease in urban New South Wales. DESIGN: Prospective laboratory surveillance. SETTING: Microbiology laboratories and hospitals in the Sydney, Hunter and Illawarra Statistical Divisions of NSW, June 1997 to May 1999. RESULTS: 1270 cases were identified in two years. Incidence of disease was highest in those aged < 2 years (96.4 per 100,000; 95% CI, 83.7-107.9) and > or = 85 years (100.1 per 100,000; 95% CI, 81.8-121.3). Incidence of disease increased significantly from the age of 60 years, compared with low rates in those aged 5 59 years. Underlying diseases predisposing to pneumococcal infection increased with age, from 4% (< 2 years) to 60% (> or = 65 years). A seven-valent conjugate vaccine would have covered 84.8% of serotypes in those aged 0-14 years, falling to 69% in those > or = 15 years. Penicillin resistance was significantly higher in the < 5 years group (19.0%) than in older people (14.6%). CONCLUSIONS: Incidence of invasive pneumococcal disease was higher in this study using active surveillance than in previous Australian studies. An effective sevenvalent conjugate pneumococcal vaccine could prevent more than 80% of cases in children aged < 5 years. PMID- 11062803 TI - Invasive pneumococcal disease in the Northern Territory of Australia, 1994-1998. AB - OBJECTIVE: To examine the epidemiology of invasive pneumococcal disease (IPD) in the Northern Territory of Australia as a basis for optimising vaccination and healthcare provision. DESIGN: Prospective laboratory surveillance, with information collected from hospital and clinic records. SETTING: Northern Territory (NT) and rural communities in north-west South Australia served by an NT hospital, 1994-1998 (NT population is 27% Indigenous). MAIN OUTCOME MEASURES: IPD incidence and mortality, risk factors, clinical presentation and disease causing serotypes in Indigenous and non-Indigenous people. RESULTS: 425 cases of IPD were detected, with 77% in Indigenous people. IPD incidence was highest in Indigenous children aged < 2 years (1534 per 100,000 in central Australia), but about 100 per 100,000 in non-Indigenous children < 2 years and all Indigenous age groups aged > or = 15 years. Mean ages of those with disease were 39 years in Indigenous people and 48 years in non-Indigenous people (P = 0.006) and, of those who died, 41 and 53 years, respectively (P = 0.04). IPD risk factors were present in 72% of Indigenous and 55% of non-Indigenous patients aged > or = 2 years. Serotype results for 363 isolates showed that the 23-valent vaccine covered 68% and 85% of isolates from Indigenous and non-Indigenous people aged > or = 2 years, respectively, while the proposed seven-, nine- and 11-valent conjugate vaccines covered 58%, 66% and 67% of isolates, respectively, from Indigenous children aged < 2 years and 72% each of those from non-Indigenous children. Case fatality rates were 10% in both Indigenous and non-Indigenous people. CONCLUSION: These data support the recent change in NT vaccination policy which extended funding for the 23-valent vaccine to all Indigenous people aged > or = 15 years and all Indigenous children in central Australia aged 2-5 years. The high rates of IPD in both Indigenous and non-Indigenous children mandate action to make conjugate vaccine available as soon as possible. PMID- 11062804 TI - Invasive pneumococcal disease in the population of Victoria. AB - OBJECTIVES: To estimate morbidity and mortality rates for invasive Streptococcus pneumoniae (pneumococcal) disease in the non-Indigenous population of Victoria. DESIGN AND SETTING: Survey using data from a statewide voluntary laboratory surveillance scheme (1989-1998), statewide hospital discharge database (1995 1998), medical records of notified patients (1994-1995) and serotyping of notified isolates (1994-1998). MAIN OUTCOME MEASURES: Incidence of pneumococcal bacteraemia and pneumonia; predisposing factors; serotypes of isolates. RESULTS: Minimum estimates of annual incidence of invasive disease, based on laboratory surveillance data for 1995-1998, were 59 per 100,000 for children aged < 2 years, 25 per 100,000 for people aged > or = 65 years, and 8 per 100,000 overall. Annual incidence of pneumococcal pneumonia, calculated from hospital discharge data, was 99 per 100,000 for those aged > or = 65 years. Manifestations of invasive pneumococcal disease varied with age, with meningitis more common in infants, and pneumonia most common in older patients. A predisposing factor for pneumococcal infection was present in 48% of patients. Most isolates from infants (83%) belonged to serotypes in the proposed seven-valent infant vaccine, and 91% of isolates from people aged > or = 2 years belonged to serotypes in the current 23 valent adult vaccine. CONCLUSIONS: S. pneumoniae continues to be a major cause of morbidity and mortality in young children and the elderly in Victoria. More widespread use of the currently available pneumococcal vaccine in adults and introduction of an effective vaccine for infants should greatly reduce incidence of the disease. PMID- 11062805 TI - Are current recommendations for pneumococcal vaccination appropriate for Western Australia? The Vaccine Impact Surveillance Network--Invasive Pneumococcal Study Group. AB - OBJECTIVE: To examine the epidemiology of invasive pneumococcal disease (IPD) in Western Australia (WA), and thereby to assess whether current Australian recommendations on vaccination are suitable for WA and whether vaccines cover the local IPD-causing serotypes. DESIGN AND SETTING: Prospective laboratory surveillance, with data gathered on all people in WA who had Streptococcus pneumoniae (pneumococcus) isolated from a normally sterile body site, 1 April 1996 to 1 April 1997. MAIN OUTCOME MEASURES: IPD incidence and mortality rates, risk factors, clinical presentation and disease-causing serotypes. RESULTS: IPD incidence and mortality were higher in the Indigenous than non-Indigenous population (relative risk [RR] for incidence, 11.9; 95% CI, 6.8-14.9, P < 0.001; RR for mortality, 22.7; 95% CI, 7.2-71.3, P < 0.001), but the case-fatality rate did not differ significantly. Incidence increased significantly from the ages of 25 years in Indigenous people and 60 years in non-Indigenous people. Most disease causing isolates from adults (94% from non-Indigenous and 86% from Indigenous adults) belonged to serotypes included in the current 23-valent vaccine. Thirty one isolates (92%) from non-Indigenous children (aged < 15 years) belonged to serotypes included in the new seven-valent conjugate vaccine, compared with only five (50%) from Indigenous children (chi 2 = 9.7; P < 0.001). CONCLUSIONS: Current Australian vaccination recommendations do not appear to match IPD epidemiology in WA; consideration should be given to lowering the age criterion for vaccination to 25 years for Indigenous and 60 years for non-Indigenous people. The new seven-valent conjugate vaccine may not adequately cover disease causing serotypes in Indigenous children. PMID- 11062806 TI - Efficacy and effectiveness of pneumococcal polysaccharide vaccines and their use in industrialised countries. AB - Use of pneumococcal polysaccharide vaccines remains controversial, even though clinical trials have shown their efficacy in preventing invasive disease caused by vaccine serotypes in immunocompetent adults. Retrospective studies indicate these vaccines have about 50%-80% effectiveness for preventing invasive disease caused by vaccine serotypes, although effectiveness wanes over time and with age. The elderly, people living in institutions and those with chronic cardiac or respiratory disease, alcoholism or diabetes mellitus who are in relatively good health would benefit from vaccination; a polysaccharide vaccine program in the elderly has been shown to be cost-effective. In young children, polysaccharide vaccine should be evaluated as a booster to conjugate pneumococcal vaccines, which are likely to be available soon in industrialised countries. In view of the high rates of hospitalisation and of antimicrobial resistance in pneumococci, every effort should be made to increase coverage by pneumococcal polysaccharide vaccine in Australia, according to guidelines of the National Health and Medical Research Council. PMID- 11062807 TI - Improving pneumococcal vaccination coverage among older people in Victoria. AB - Although pneumococcal vaccine is recommended by the National Health and Medical Research Council and is cost-effective in preventing invasive pneumococcal disease, it is the only vaccine on the standard schedule that is not nationally funded through public health grants to the States. In Victoria, the Department of Human Services has provided free pneumococcal vaccine to people aged 65 years and over since 1998. Pneumococcal vaccination was given in conjunction with the annual influenza vaccination program; 28.5% of the eligible cohort (95% CI, 24.8% 32.1%) received pneumococcal vaccine in 1998, giving an estimated cumulative coverage of 42% (13.4% had received it in 1997). We expect coverage will continue to increase over time, but revaccination every five years will present a substantial financial burden; access to vaccine is critical to improving coverage. Our experience in Victoria suggests that a nationally funded program, administered similarly to the influenza vaccination program, would dramatically increase pneumococcal vaccination coverage at a national level. PMID- 11062808 TI - Conjugate pneumococcal vaccines: an overview. AB - A seven-valent conjugate pneumococcal vaccine has been shown to have dramatic efficacy against invasive pneumococcal disease and lesser efficacy against otitis media and pneumonia. This vaccine was licensed for use in infants in the United States in February 2000 and is recommended there for routine use in infants and catch-up vaccination in high-risk children. Specific regional pneumococcal vaccines are not needed; nine to 11 serotypes cover most pneumococcal disease in most parts of the world; nine- and 11-valent conjugate vaccines are currently being developed for the global market. There is evidence of serotype replacement in vaccine recipients (in both carriage and disease), which might reduce overall vaccine effectiveness. There is also evidence that vaccines may reduce rates of antimicrobial resistance in pneumococci. Studies of the burden of pneumococcal disease as well as program support are needed to assist developing countries to introduce these expensive vaccines. PMID- 11062809 TI - Conjugate pneumococcal vaccines for aboriginal children in Australia. AB - Research indicates a high burden of pneumococcal disease and great potential benefits of conjugate vaccines in Indigenous Australian children, who should have high priority for delivery of these vaccines. Incidence of invasive pneumococcal disease in Indigenous people in central Australia is the highest reported in the world (2053 per 100,000 persons per year in those aged under two years). Acute respiratory infection is a major cause of morbidity in Indigenous children in rural and remote areas. Early pneumococcal colonisation of the nasopharynx and high rates of carriage are seen in Indigenous children, and are probably related to their high rates of ear disease. Current seven-valent conjugate vaccines are likely to cover about two-thirds of invasive isolates in Indigenous Australian children; 11-valent vaccines will cover a higher proportion. Questions remain about the best vaccine carrier protein and the likely impact of vaccine on ear disease, pneumococcal carriage and antibiotic resistance. PMID- 11062810 TI - Conjugate pneumococcal vaccines for non-indigenous children in Australia. AB - Childhood pneumococcal disease is associated with substantial morbidity and mortality, but total disease burden is more difficult to measure than for invasive disease caused by Haemophilus influenzae type b (Hib). A safe, effective seven-valent conjugate pneumococcal vaccine will be available in Australia by early 2001, and will certainly be indicated for high-risk groups and purchased in the private sector, as was Hib vaccine. The status of this vaccine on the Australian Standard Vaccination Schedule will require more detailed consideration of the burden and serotype distribution of pneumococcal disease in Australian children and the vaccine's likely cost-effectiveness. Postmarketing surveillance will be particularly important. PMID- 11062811 TI - Antibiotic resistance in Streptococcus pneumoniae. AB - Resistance to penicillin and other antibiotics in Streptococcus pneumoniae has emerged in Australia and around the world in the past decade, and appears to be worsening (e.g., rates of penicillin resistance in Australia rose from 1% in 1989 to 25% in 1997). In Australia, the only oral antibiotic able to treat respiratory infections caused by some multiresistant strains is high-dose amoxycillin. If these strains increase in prevalence, then treatment failures for relatively minor infections (e.g., otitis media) are likely to become common, resulting in repeat antibiotic courses or hospitalisation for parenteral therapy. Therapy for meningitis caused by penicillin-sensitive pneumococcal strains remains high-dose benzylpenicillin, but empirical treatment while awaiting culture and sensitivity results is problematic; neither penicillin nor third-generation cephalosporins cover all strains. Therefore, many authorities recommend vancomycin, usually combined with a third-generation cephalosporin, for treating presumptive or proven pneumococcal meningitis pending penicillin-susceptibility results. As almost all readily available oral antibiotics in Australia select for resistant strains of pneumococci, multiresistant strains will increase in prevalence unless unnecessary antibiotic use and prescription volumes are reduced substantially in the next few years. PMID- 11062812 TI - The case for doubling investment in biomedical research. PMID- 11062813 TI - Enhanced surveillance of HIV infections in New Zealand, 1996-1998. AB - AIM: To improve understanding of the HIV epidemic in New Zealand through use of an enhanced voluntary reporting system for new diagnoses of HIV. METHODS: Routine reporting of new HIV diagnoses by the two laboratories that perform confirmatory HIV antibody testing, to the Department of Health and later to the AIDS Epidemiology Group, has been in place since 1985. From January 1996, this was supplemented by a questionnaire about demographic characteristics and circumstances of HIV exposure sent to clinicians requesting the HIV test. RESULTS: From January 1996 to December 1998, 260 new diagnoses of HIV were reported (205 males, 55 females) and extra information was obtained from clinicians for 253 (97.3%) people. HIV diagnosis rate was highest for 'other' ethnicity and similar for European, Maori and Pacific Island ethnic groups. Sexual intercourse between men was the commonest mode of infection (43.5%), followed by heterosexual intercourse (40.0%) and injecting drug use (2.7%). Places of infection were New Zealand (38.5%), Australia (7.7%), 'other' overseas (45.4%) and unknown (8.5%). Heterosexual infections were acquired through contact with a person in or from a high prevalence area (mainly in Africa or Asia) for 86.7% of males and 68.2% of females. Second generation heterosexual transmission was rare. CONCLUSIONS: Introduction of an enhanced surveillance system has been successful. Results confirm continuing spread of HIV in New Zealand amongst men who have sex with men, and suggest low levels of heterosexual and injecting drug use transmission in New Zealand. Of major importance in the occurrence of heterosexual infection is the role of imported HIV. PMID- 11062814 TI - The prevalence of viral hepatitis (HAV, HBV and HCV) in the Christchurch community. AB - AIM: To determine the prevalence of hepatitis A (HAV), hepatitis B (HBV) and hepatitis C (HCV) in adults randomly selected from the Christchurch community. METHODS: A list of names was randomly generated from the Christchurch electoral roll and subjects were sequentially contacted and invited to participate. A blood sample was taken and tested for hepatitis A (IgG anti-HAV antibody), hepatitis B (HBsAg and anti-HBc) and HCV (anti-HCV antibody) using Abbott Elisa kits. Subjects positive for HBsAg were also tested for HBeAg/HBV DNA. Those positive for anti-HBc were tested for anti-HBs. HCV antibody positive samples were tested for HCV RNA using PCR. RESULTS: 1064 subjects (30.3% of those invited) participated in the study. The prevalence of HAV antibodies was 27.9%, and increased with age. The overall prevalence of HBV markers was 42/1064 (4.2%), and of these 0.3% were HBsAg positive and 3.9% were considered immune. No gender or ethnic differences in these proportions were observed. The seroprevalence of HVC antibody was 3/1064 (0.3%), two of whom were also PCR positive for HCV RNA. CONCLUSION: In the Christchurch community there was a high prevalence of antibodies to HAV, which increased with age. The prevalence of HBsAg and antibody to HCV were both low at 0.3%. PMID- 11062815 TI - Users of unconventional practitioners: a profile of 26 year old New Zealanders. AB - AIMS: To profile 26 year old New Zealand users of unconventional practitioners. METHODS: 977 members of the Dunedin Multidisciplinary Health and Development Study participating in the age-26 assessment (1998-1999) answered questions about twelve-month service use, education, income, recent medical history, current health status and avoidance of medical situations. RESULTS: 10% had used an unconventional practitioner in the previous twelve months. The majority (88%) had also used a conventional practitioner. Those using both types of practitioner were heavy users of health services (twelve visits/year). Compared to those who used conventional practitioners exclusively, they had significantly higher incomes and were more likely to report a serious injury, a current disability, a history of back problems, role limitations due to physical health problems, and more bodily pain (all p < 0.01). CONCLUSIONS: 26 year old New Zealand users of unconventional practitioners have a similar profile to their counterparts in other developed countries. It appears that their health needs are not fully met by conventional services, emphasising the need for more research into the aetiology and treatment of ailments (e.g. back pain) for which unconventional practitioners are commonly sought. The Medical Council of New Zealand guidelines on unconventional medicine are discussed in light of these findings. PMID- 11062816 TI - Funding of biomedical research in New Zealand. PMID- 11062817 TI - Retention of junior medical officers. PMID- 11062818 TI - 'OOS' and ergonomic considerations. PMID- 11062819 TI - Doctors as managers. PMID- 11062820 TI - Doctors as managers. PMID- 11062821 TI - Pacifier use and morbidity. PMID- 11062822 TI - Iron and athletic performance. PMID- 11062823 TI - Penetrating intracranial trauma in an infant secondary to a modified baby bouncer. PMID- 11062824 TI - Gastrointestinal decontamination following paraquat ingestion. PMID- 11062825 TI - Job satisfaction, rural practice and the health reforms. PMID- 11062826 TI - Noise levels. PMID- 11062827 TI - Late recurrence in paediatric melanoma. PMID- 11062828 TI - Bridging the gap. PMID- 11062829 TI - Shouldn't Canadian veterinary colleges be providing training in veterinary dentistry? PMID- 11062830 TI - Understanding mechanism of action versus alleviating suffering. PMID- 11062831 TI - An ethicist's commentary on research protocols requiring significant animal suffering. PMID- 11062832 TI - Ten tips to ease the transition from student to veterinarian. PMID- 11062833 TI - The case against alternative medicine. PMID- 11062834 TI - Observations on topical ivermectin in the treatment of otoacariosis, cheyletiellosis, and toxocariosis in cats. AB - The purpose of this study was to observe the efficacy of a topical pour-on formulation of ivermectin in the treatment of otoacariosis, cheyletiellosis, and toxocariosis in cats. Forty-five cats were treated. All cats received 2 to 4 topical applications of ivermectin on the skin between the shoulder blades in a narrow strip, 14 days apart. This practical treatment was effective in 96% (23/24) of cases of feline otoacariosis and in 100% (20/20) of cats with toxocariosis. All cats with cheyletiellosis (16/16) received 4 treatments and had resolution of clinical signs, but one Cheyletiella egg could still be found 45 days after the last treatment. The viability of this egg could not be evaluated, but the cats were still free of clinical signs on follow-up 6 months later. The treatment was well tolerated in all the animals. A few cats developed a transient small alopecic area and mild scaling at the site of application of the drug. PMID- 11062835 TI - Etiology, forms, and prognosis of gastrointestinal dysfunction resembling vagal indigestion occurring after surgical correction of right abomasal displacement. AB - The objectives of this study were to determine the etiology and types of vagal indigestion (VI) occurring after right displacement of the abomasum or abomasal volvulus (RDA/AV), and the prognosis for each type. Data of cows presented for RDA/AV from a retrospective (n = 288) and a prospective (n = 132) study were used. Vagal indigestion occurred in 39 and 22 cows in each study, respectively. A necropsy was performed in 29 cases. Gastric compartment dilation compatible with VI type III or IV occurred in 23 cases. An abnormal gastric wall was detected in 22 cases. Peritonitis was present in 18 cows. Vagal nerve lesions were present in 5 out of 13 cases studied. Clinical, hematological, and necropsy results suggested a classification of VI with respect to presence or absence of peritonitis. Gastric wall damage, peritonitis and vagal nerve lesions appear important in the etiology. Considering peritonitis occurrence, antimicrobial therapy appears necessary in the treatment of RDA/AV. PMID- 11062836 TI - Estrus synchronization and pregnancy rates in beef cattle given CIDR-B, prostaglandin and estradiol, or GnRH. AB - Two experiments were conducted to determine estrous response and pregnancy rate in beef cattle given a controlled internal drug release (CIDR-B) device plus prostaglandin F2 alpha (PGF) at CIDR-B removal, and estradiol or gonadotropin releasing hormone (GnRH). In Experiment I, crossbred beef heifers received a CIDR B device and 1 mg estradiol benzoate (EB), plus 100 mg progesterone (E + P group; n = 41), 100 micrograms gonadotropin releasing hormone (GnRH group; n = 42), or no further treatment (Control group; n = 42), on Day 0. On Day 7, CIDR-B devices were removed and heifers were treated with PGF. Heifers in the E + P group were given 1 mg EB, 24 h after PGF, and then inseminated 30 h later. Heifers in the GnRH group were given 100 micrograms GnRH, 54 h after PGF, and concurrently inseminated. Control heifers were inseminated 12 h after onset of estrus. The estrous rate was lower (P < 0.01) in the GnRH group (55%) than in either the E + P (100%) or Control (83%) groups. The mean interval from CIDR-B removal to estrus was shorter (P < 0.01) and less variable (P < 0.01) in the E + P group than in the GnRH or Control groups. Pregnancy rate in the E + P group (76%) was higher (P < 0.01) than in the GnRH (48%) or Control (38%) groups. In Experiment II, 84 cows were treated similarly to the E + P group in Experiment I. Cows received 100 mg progesterone and either 1 mg EB or 5 mg estradiol-17 beta (E-17 beta) on Day 0 and either 1 mg of EB or 1 mg of E-17 beta on Day 8 (24 h after CIDR-B removal), in a 2 x 2 factorial design, and were inseminated 30 h later. There were no differences among groups for estrous rates or conception rates. The mean interval from CIDR-B removal to estrus was 44.2 h, s = 11.2. Conception rates were 67%, 62%, 52%, and 71% in Groups E-17 beta/E-17 beta, E-17 beta/EB, EB/E-17 beta, and EB/EB, respectively. In cattle given a CIDR-B device and estradiol plus progesterone, treatment with either EB or E-17 beta effectively synchronized estrus and resulted in acceptable conception rates to fixed-time artificial insemination. PMID- 11062837 TI - A study of the inheritance of a bleeding disorder in Simmental cattle. AB - A study was designed to determine the inheritance pattern of a blood platelet aggregation disorder in Simmental cattle utilizing embryo transfer technology. A Simmental donor cow that had previously produced a calf with the platelet aggregation disorder was superovulated and mated to a bull that had also produced affected offspring. Twenty-seven calves were produced from the 63 (42.9%) embryos transferred. This somewhat lower than expected pregnancy rate is suggestive of an increased rate of embryo loss. Twenty-three of 25 (92%) calves had normal platelet aggregation patterns and 2 failed to show any evidence of platelet aggregation. Data are suggestive that inheritance is not simple Mendelian recessive. A more likely scenario is that the defect is the result of the inheritance of at least 2 genes, which is also consistent with the sporadic incidence reported in the population at large. PMID- 11062838 TI - Urinary bladder transitional cell papilloma in a shorthorn heifer. AB - A 12-month-old shorthorn heifer was presented for pollakiuria of 4 months' duration. Urinary bladder transitional cell papilloma was diagnosed. The heifer had no exposure to bracken fern and no papillomavirus or bacterium was demonstrated. Laser surgery was used in an attempt to debulk the mass. PMID- 11062839 TI - Toxigenic and drug resistance properties of porcine Pasteurella multocida isolates from Prince Edward Island. PMID- 11062840 TI - A mixed fungal infection in a dog: sporotrichosis and cryptococcosis. AB - Unusual ulcerated masses protruding from both nostrils of a 3-year-old terrier were diagnosed histologically as sporotrichosis, and regressed with iodide therapy. Cryptococcus neoformans was recovered from new lesions that appeared near the dog's eye and on the extremities. All lesions regressed with itraconazole therapy. PMID- 11062841 TI - Staff turnover or staff retention: understanding the dynamics of generations at work in the 21st century. PMID- 11062842 TI - Alessandro Agnoli Lecture. Models of basal ganglia dysfunction: predictions and pitfalls. PMID- 11062843 TI - Functional and clinical changes in upper limb spastic patients treated with botulinum toxin (BTX). AB - Spasticity is a motor disorder characterized by a velocity-dependent increase in tonic stretch reflexes (muscle tone) with exaggerated tendon jerks. In order to study the usefulness of botulinum toxin type A (BTX) as a therapy for spasticity, we studied 15 patients affected by spasticity secondary to stroke. Tests included: clinical evaluation of tone (Ashworth scale); active angles of extension and flexion at elbow and wrist; Hmax/Mmax ratio from flexor carpi radialis (FCR); Hreflex presynaptic inhibition from FCR during vibration; Task score; and video recording. Patients were injected with BTX into one or more muscles with total doses not exceeding 200 International Units (IU). The tests were performed immediately prior to injection and repeated 2 weeks afterwards. Furthermore, in eight patients, testing was also performed one month after BTX injection. Between two weeks and one month after BTX there were no statistically significant differences. A statistically significant difference in the Task and Ashworth scores before and after treatment emerged (p < 0.0014), but only 6 patients showed a clear improvement in motor performance. Overall, we observed an improvement in the angle of active extension and flexion at the wrist and elbow. There were no significant changes in the Hmax/Mmax ratio and the Hreflex presynaptic inhibition during vibration. All the patients reported a subjective improvement. The results suggest that subjective benefits can be gained from the use of BTX in patients affected by spasticity, and that the degree of motor improvement seems to depend on the motor recovery obtained before treatment. PMID- 11062844 TI - Long-term clinical and electrophysiological results of local steroid injection in patients with carpal tunnel syndrome. AB - The aim of this study was to evaluate the effects of local steroid injection on both the clinical symptoms and motor and sensory conduction of the median nerve in carpal tunnel syndrome (CTS). Using a standard evaluation and treatment protocol, we prospectively studied steroid injection in 32 hands in 24 patients (mean age: 50.7 +/- 10 years; 23 women and 1 man). To determine the normal median nerve values, 42 normal controls (mean age: 39.1 +/- 10.5 years; 21 women and 21 men) were also studied. At follow-up, clinical symptom scores and signs of CTS, as well as electrophysiological variables of the median nerve, showed a significant trend towards improvement with respect to baseline values (p < 0.01). By the end of the one-year follow-up period, the symptoms had remitted completely or partially in 27 hands (84.4%). In addition to the relief of the symptoms, motor nerve conduction abnormalities had improved in 62.6% of hands, and anti dromic sensory nerve conduction abnormalities in 62.5%. In cases where a sensory nerve action potential (SNAP) could be obtained, the efficacy of this treatment was found to be relatively higher than in cases in which SNAP was absent (p < 0.01). PMID- 11062845 TI - Sumatriptan overuse in episodic cluster headache: lack of adverse events, rebound syndromes, drug dependence and tachyphylaxis. AB - This observational study was designed to examine the pattern of sumatriptan use in patients with cluster headache using more than the recommended daily dose of subcutaneously injected (s.c.) sumatriptan. Thirteen patients suffering from episodic cluster headache were asked to record the characteristics of their attacks and drug intake for 1 year. All reported a high daily frequency of attacks (more than 3 per day) and the related overuse of s.c. sumatriptan. The results show that the overall incidence of adverse events among patients receiving sumatriptan injections for the treatment of cluster headache is low. The extended administration of this drug in episodic cluster headache did not result in tolerance problems or tachyphylaxis. Only 4 patients experienced minor adverse events and recovered more slowly than the others. They suffered from migraine without aura and cluster headache, and showed a family history of migraine. Even though they must be viewed with caution, due to the observational nature of the study and the low number of patients included, these results suggest that the profile of sumatriptan may differ in cluster headache compared with migraine. PMID- 11062846 TI - Energy consumption and gait analysis in children with myelomeningocele. AB - The aim of this study was to determine, in children with different levels of myelomeningocele (MMC), the gait pattern and energy cost of walking with and without ankle-foot orthoses (AFOs). We found that each MMC level was characterised by recognisable gait patterns and that the abnormalities closely reflected the muscle deficits present. Furthermore, the study also introduces new indices for evaluating the energy cost of locomotion and demonstrates that the energy required for walking is increased in children with MMC compared with non disabled children. With respect to barefoot conditions, the use of AFOs leads to an improvement in gait and reduced energy consumption. PMID- 11062847 TI - Genotype-phenotype correlation in familial amyotrophic lateral sclerosis with SOD1 mutation. PMID- 11062848 TI - Routine blastocyst culture and transfer: 201 patients' experience. AB - PURPOSE: The purpose was to compare the outcomes between day-5 blastocyst and day 2 embryo transfers. METHODS: Infertile women who accepted the ovarian hyperstimulation and oocyte retrieval were divided: Group 1, day 2 embryo transfer, group 2, cultured to day 5 in serum-free sequential culture medium and transfer. Early embryo quality and growth, blastocyst formation and quality, implantation rate (IR) and pregnancy rates (PR) were detected. RESULTS: Total blastocyst formation rate was 49.4%. Better early embryo quality (days 2, 3) and higher day 3 blastomere number possess higher blastocyst formation rate. The IR for day 2 and day 5 embryos were 10.8% and 22.2%, respectively. The PR in both groups were comparable (37.3% vs. 41.8%). CONCLUSIONS: Blastocyst transfer has higher IR and comparable PR as those of day 2 embryo transfer. Early embryo qualities and day 3 blastomere number are useful in predicting the final blastocyst formation. Blastocyst formation rate is not related to maternal age, infertile causes, insemination methods, and early embryo number. PMID- 11062849 TI - Prediction of pregnancy rate of in vitro fertilization and embryo transfer in women aged 40 and over with basal uterine artery pulsatility index. AB - PURPOSE: The purpose was to determine the effect of basal uterine perfusion on the pregnancy rates of in vitro fertilization and embryo transfer (IVF-ET) in women aged 40 and above. METHODS: A total of 47 patient aged 40 and over underwent IVF-ET. The conception cycles and the nonconception cycles were compared. RESULTS: Of the 47 patients, 4 patients were pregnant (8.5%). The mean age, basal follicle stimulating hormone (FSH), basal estradiol (E2) level, antral follicle count (AFC), number of ampoules of gonadotropin used, E2 levels and endometrial thickness on the day of human chorionic gonadotropin (hCG) administration, number of retrieved and fertilized oocytes, and number of transferred embryos were not statistically significant between the conception and nonconception cycles. However, the basal uterine artery pulsatility index (UA PI) was significantly lower in the conception cycles (P < 0.001). The receiver operating characteristics (ROC) curve analysis for basal FSH, AFC, and basal UA PI in predicting the pregnancy rate of IVF in patients aged > or = 40 were demonstrated. The best prediction rate was achieved by a pulsatility index cutoff of < 2.0 for a receptive uterus. CONCLUSIONS: Increased uterine perfusion in the early follicular phase enhanced the pregnancy rate of IVF in women aged 40 and above. It is therefore essential that patients aged > or = 40 with poor basal uterine perfusion should be identified early in the early follicular phase of the menstrual cycle to apply appropriate intervention to improve the uterine circulation for the subsequent chance of pregnancy. PMID- 11062850 TI - Leukemia inhibitory factor expression in different endometrial locations between fertile and infertile women throughout different menstrual phases. AB - PURPOSE: The purpose was to demonstrate the leukemia inhibitory factor (LIF) expression in different endometrial locations between fertile and infertile women throughout different menstrual phases. The relationship between progesterone level and LIF expression were evaluated. METHODS: Endometrial biopsies were performed on idiopathic infertile and normal fertile women accepted the in follicular, periovulatory, and luteal phases. The luteal progesterone level was measured. Endometrial LIF immunostaining of luminal epithelium, glandular epithelium, and stroma were detected. The relationship between luteal LIF expression and progesterone level was evaluated. RESULTS: Significant LIF expression was noted in the endometrium of fertile women rather than that of infertile women. The LIF expression was highest in the luminal epithelium, moderate in the glandular epithelium, and lowest in the stroma. The luminal and glandular epithelial staining were lowest in follicular phase, moderate in periovulatory phase, and strongest in luteal phase. The stromal LIF presented with a noncyclical manner. The LIF expression is not related with the progesterone level. CONCLUSIONS: Endometrial LIF expression is related to human fertility. Endometrial LIF expression is dependent on cellular localizations and menstrual stages. Stronger LIF expression presents in the endometrial epithelium during luteal phase. PMID- 11062851 TI - Late fertilization of unfertilized human oocytes in in vitro fertilization and intracytoplasmic sperm injection cycles: conventional insemination versus ICSI. AB - PURPOSE: The aim of this study was to evaluate the efficacy of intracytoplasmic sperm injection (ICSI) in comparison with conventional reinsemination using fertilization failed oocytes by conventional in vitro fertilization (IVF). METHODS: Oocytes were collected from patients of IVF or ICSI cycles. Patients were grouped by fertilization techniques: group 1: conventional IVF; group 2: reinsemination after conventional IVF failure; group 3: regular ICSI; group 4: 1 day-old ICSI after conventional IVF failure; group 5: 2-day-old ICSI after conventional IVF failure; group 6: re-ICSI after regular ICSI failure. RESULTS: In different insemination groups, normal fertilization rate was higher (P < 0.001) in 1-day-old ICSI (47.1%) and 2-day-old ICSI groups (40.0%) than in reinsemination (14.7%). Abnormal fertilization rate was higher (P < 0.05) in re ICSI group (21.7%) than any other groups (range: 0-8%). Cleavage rate was higher in 1-day-old (36.7%) and 2-day-old ICSI groups (36.0%) than in reinsemination (5.3%, P < 0.001) or re-ICSI groups (17.4%, P < 0.05). Pregnancy rate was 27.6% and 20.0% in conventional IVF and regular ICSI groups, respectively. However, 1 day-old ICSI (group 4) and 2-day-old ICSI (group 5) were attempted once embryo transfer (ET) but failed pregnancy occurred in each group. CONCLUSIONS: In fertilization failure cycles, late ICSI increases the rate of fertilization and embryonic development and may rescue the completely failed attempt of pregnancy. PMID- 11062852 TI - Oocyte morphology and early zygote cleavage does not appear to be related to intrafollicular concentrations of inhibin-A or -B. AB - PURPOSE: The aim was to investigate the relationship of intrafollicular inhibin dimers A and B with human oocyte morphology and subsequent embryo potential. METHODS: Sixty-eight oocytes were isolated from 31 women undertaking intracytoplasmic sperm injection (ICSI). Estradiol, progesterone, testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin, sex hormone-binding globulin, inhibin-A and inhibin-B was assayed in corresponding follicular fluid. RESULTS: The mean (+/- SD) concentration for inhibin-A was 9.7 +/- 9.8 ng/ml (range, 1.1-60.0 ng/ml) and for inhibin-B was 269.4 +/- 185.2 ng/ml (range, 33.1-811.0 ng/ml). In a correlation matrix there were no marked relationships (r < 0.556) between inhibin and steroid or gonadotropin concentrations. Similarly, when inhibin concentrations were divided according to whether the oocytes had mature or immature cumulous complexes, were viable or necrotic, were meiotically immature or mature, became fertilized or not, or had different embryo gradings after cleavage, no statistically significant difference could be seen between groupings. CONCLUSIONS: Because the range of values was large and the data often skewed, neither inhibin dimer has discriminatory power to reflect the potential of the oocyte. PMID- 11062853 TI - Very low sperm count affects the result of intracytoplasmic sperm injection. AB - PURPOSE: The aim was to examine the influence of extremely low sperm count on intracytoplasmic sperm injection (ICSI) outcome. METHODS: Over 1000 consecutive unselected ICSI cycles were divided into four groups according to sperm concentration of their patients: A, cryptozoospermia, 107 patients; B, sperm concentration of < or = 1 x 10(4), 146 patients; C, sperm count of 1 x 10(4)-1 x 10(5), 135 patients; and concentration of > 1 x 10(5) and < 10 x 10(6)/ml (control group), 688 patients. RESULTS: A significant decrease in pregnancy rate was noticed in the cryptozoospermic group in comparison to the control group (20% vs. 31%). Fertilization rate in group A was significantly lower in comparison to all other groups, respectively (46% vs. 52%, 54%, 61%). Embryo quality was inferior in group A in comparison to the control group. A higher yet not statistically significant abortion rate was observed in the cryptozoospermic group (as well as in group C) (30%, 27%) compared to the control group (15%). CONCLUSIONS: It seems that an extremely low sperm count has a negative effect on the outcome of ICSI. Nevertheless patients with cryptozoospermia should not be offered ICSI treatment with the ejaculated sperm before karyotype is established. PMID- 11062854 TI - Increased progesterone secretion and 3 beta-hydroxysteroid dehydrogenase activity in human cumulus cells by pregnenolone is limited to the high steroidogenic active cumuli. AB - PURPOSE: Several reports imply that lower progesterone secretion by cumulus oocyte complexes (COCs) is associated with lower fertilization in the corresponding oocyte. The possible role of progesterone in oocyte fertilization in humans was studied using two approaches: (a) increasing the total progesterone secretion by culturing more than one COC per dish; and (b) increasing the cumulus cell progesterone secretion by providing pregnenolone as a substrate. METHODS: Mature COCs were cultured individually or cocultured in groups. Oocyte fertilization and progesterone secretion were tested after 20 hr and 3 days in culture, respectively. The cumuli from individually plated COCs were cultured in the absence of oocyte for an additional 3 days in order to test the effects of pregnenolone on progesterone secretion and the 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity. A comparable study with pregnenolone was performed on the corresponding granulosa-lutein cells. RESULTS: Increasing the number of COC to two instead of one led to a significant increase in both fertilization rate and progesterone secretion. The addition of pregnenolone during days 3-6 increased significantly both progesterone secretion and 3 beta HSD activity. Comparable results were observed in granulosa-lutein cells subjected to pregnenolone treatment. Following the first 3 days culture, cumulus masses were categorized as secreting high or low progesterone levels. Adding pregnenolone had a greater effect on both progesterone secretion and 3 beta-HSD activity in the high-progesterone-secreting cumuli. CONCLUSIONS: Addition of pregnenolone increased progesterone secretion and 3 beta-HSD more efficiently in the higher-progesterone-secreting cumuli. Coculture of two COCs instead of one led to a higher fertilization rate and greater progesterone secretion. PMID- 11062855 TI - A study on psychological strain in IVF patients. AB - PURPOSE: The objectives of this study were to compare average stress levels in infertile women to fertile women, to determine the stress levels whether the patients was pregnant or not pregnant, and to examine for a cross-section of infertile patients in different stages of medical investigation for the infertility. METHODS: One hundred thirty-eight women receiving medical treatment for infertility attended the program. The State Trait Anxiety Inventory (STAI) and the Beck Depression Inventory (BDI) of perceived stress associated with the infertility was the outcome measure. RESULTS: Infertile women showed significant increases in trait anxiety and depressive symptoms than the fertile women. Anxiety and depression in the in vitro fertilization (IVF)-failed women were significantly higher than the IVF-success women. According to the duration of infertility, STAI and BDI were moderately elevated in the first stage (< 3 year). There was a trend of a decreasing psychological stress with an advanced infertility duration. On depression scales, the intermediate and final duration of infertility patients showed less symptomatology than the first-stage patients. Contrary to the expectation, demographic factors such as religion and husband cooperation were not related to the experience of stress. CONCLUSIONS: We must pay an attention to the infertile patient, especially from the initial infertility workup. We recommend psychological counselling for IVF-failed patients. PMID- 11062856 TI - Electroejaculated baboon (Papio anubis) sperm requires a higher dosage of pentoxifylline to enhance motility. AB - PURPOSE: Sperm collected by electroejaculation often show poor motility. The objective was to determine whether the addition of the phosphodiesterase inhibitor, pentoxifylline, would stimulate electroejaculated baboon sperm motility. METHODS: Electroejaculation was performed on several occasions on a male baboon and sperm collected after familiarization. Pentoxifylline was tested at the standard concentration (1 mg/ml) and at twice the concentration. Sperm parameters were evaluated using a sperm motility analyzer, as well as acrosome and DNA integrity techniques. RESULTS: Sperm exposed to 2 mg/ml pentoxifylline had higher total motility when compared with the control and 1 mg/ml treatment. Rapid progression and velocities were higher after pentoxifylline. The acridine orange DNA normality test showed that over 90% of collected sperm had intact unfragmented DNA. About half the sperm population had normal morphology and intact acrosomes. A low percentage had cytoplasmic droplets. CONCLUSIONS: Sperm collected by rectal probe electroejaculation required a higher concentration (2 mg/ml) of pentoxifylline for enhanced total motility, rapid progression, and higher velocity. This suggested differences in membrane properties or phosphodisterase activity in electrojeaculated sperm. The electroejaculation procedure did not denature sperm DNA at the acridine orange assay level nor were the acrosomes disrupted. The present study also documented unique information on baboon kinematic parameters. PMID- 11062857 TI - Unilateral massive pleural effusion as the only principal manifestation of severe ovarian hyperstimulation syndrome. PMID- 11062858 TI - Falsely elevated follicle-stimulating hormone levels in women with regular menstrual cycles due to interference in immunoradiometric assay. PMID- 11062859 TI - Pelvic tuberculosis diagnosed by hysteroscopy during infertility evaluation. PMID- 11062860 TI - [48th scientific session of the Japanese College of Cardiology. Osaka, September 11-13, 2000. Abstracts]. PMID- 11062861 TI - [Effect of piracetam on urea-induced myoclonus in rats]. AB - Piracetam [2-oxo-1-pyrrolidineacetamide], a cyclic GABA, has been used in Europe for the treatment of patients with cognitive disorders. We investigated the effect of piracetam on urea-induced myoclonus in rats. Myoclonus was induced by intraperitoneal injection of 4.5 g/kg urea, and was recorded with EMG and video apparatus. The incidence of induced myoclonus decreased significantly by intraperitoneal injection of 300 mg/kg piracetam and oral administration of 0.3 10 mg/kg clonazepam. Furthermore, the combined application of 100 mg/kg piracetam and 0.03-0.1 mg/kg clonazepam was effective in ameliorating the myoclonus, although separate administrations were not effective. These findings suggest that piracetam is an effective drug for treating myoclonus, particularly when it is used in combination with clonazepam. PMID- 11062862 TI - [Effects of (+/-)-pindolol over increased extracellular 5-HT level induced by fluvoxamine: regional difference in effect among the raphe, dorsal hippocampus and prefrontal cortex as measured by in vivo microdialysis technique]. AB - It is known that the somatodendritic 5-hydroxytryptamine (5-HT)1A autoreceptor works to regulate the action of 5-HT neurons leading to the release of 5-HT. Our present study has addressed the possibility that (+/-)-pindolol, which is a non selective beta-adrenoceptor antagonist/somatodendritic 5-HT1A autoreceptor antagonist, might have the ability to enhance the level of extracellular 5-HT when used with selective serotonin reuptake inhibitor (SSRI; i.e., fluvoxamine). We have used freely moving rats to measure the extracellular level of 5-HT and dopamine (DA) in the raphe, dorsal hippocampus and prefrontal cortex using an in vivo microdialysis technique. Response power of (+/-)-pindolol (8 mg/kg, i.p.) to the rise in extracellular 5-HT level when used with fluvoxamine (60 mg/kg, i.p.) was significant in the raphe, dorsal hippocampus and prefrontal cortex, while the degree of augmentation was more significant in the prefrontal cortex than in the dorsal hippocampus. The extracellular level of DA when used with (+/-)-pindolol showed a tendency to decrease in the raphe while showing a tendency to increase in the dorsal hippocampus. However, no change occurred in the prefrontal cortex. This indicates that (+/-)-pindolol has the ability to block the somatodendritic 5 HT1A autoreceptors, thereby weakening the fluvoxamine-induced indirect action of the autoreceptors in the raphe. We have obtained positive result for the probability of pindolol augmentation in two regions--dorsal hippocampus and prefrontal cortex. This indicates that augmentation therapy by the combined use of fluvoxamine with 5-HT1A antagonist will be valid and effective. PMID- 11062863 TI - The effect of zolpidem and zopiclone on memory. AB - The effect of 10 mg of zolpidem (ZLP) on memory function was evaluated in healthy male adults using word recall test, passage recall test, and Sternberg's memory scanning task. This study was carried out as a double-blind cross-over study with 7.5 mg of zopiclone (ZPC) and a placebo. No difference was noted between the active drugs and placebo in the number of words recalled from the word list presented before administration. No evidence of retrograde amnesia was suggested. However, encoding ability was slightly affected as indicated by a decrease in the number of words recalled from the list presented after administration. Slight impairment of a delayed recall was noted for both of the active drugs, but the effect disappeared the next morning. In the memory scanning task, ZLP prolonged a non-specific component of reaction time 1.5 h after administration, but the effect disappeared after 12.5 h. ZPC did not prolong the reaction time. The two active drugs showed no specific effects on either memory scanning or response selection stage in short-term memory at any time. The findings suggest that residual effects did not reach clinical significance at the standard dosage, although the active drugs slightly affected encoding ability and retention soon after administration. PMID- 11062864 TI - Effect of dopaminergic drugs on the reserpine-induced lowering of hippocampal theta wave frequency in rats. AB - The effects of dopaminergic drugs on the lowering of hippocampal theta wave frequency induced by reserpine 1 mg/kg s.c. were examined. Sibutramine (monoamine reuptake inhibitor) 10 mg/kg p.o., methamphetamine (monoamine releaser) 1 mg/kg, quinpirole (dopamine D2 receptor agonist) 10 mg/kg i.p., and SKF 38393 (dopamine D1 receptor agonist) 10 mg/kg i.p. each antagonized the reserpine-induced lowering of hippocampal theta wave frequency in rats. Moreover, the combined administration of SKF 38393 1 mg/kg i.p. and quinpirole 1 mg/kg i.p. synergistically antagonized a reserpine-induced lowering of this frequency. Dosulepin, amitriptyline, and desipramine, which are weak inhibitors of dopamine reuptake, each had little effect on the reserpine-induced lowering of theta wave frequency at a dose of 40 mg/kg p.o. Furthermore, atropine (muscarinic anticholinergic drug) 20 mg/kg p.o. decreased theta wave power in the low frequency range following a shift to the lower range by reserpine. A positive correlation was observed for each of the above drugs between a reversal of reserpine-induced lowering of theta wave frequency and a reversal of impairment of reserpine-induced conditioned avoidance responses (ACAR) in rats. These results suggest that the reserpine-induced lowering of hippocampal theta wave frequency plays a role in the impairment of reserpine-induced ACAR, and that dopamine D1 and D2 receptors play important roles in antagonizing this lowering of frequency. PMID- 11062865 TI - [Preliminary report on the efficacy and safety of brief-pulse ECT in depression]. AB - Because there is no report on the use of brief-pulse devices in Japan, we have examined the efficacy and safety of brief-pulse electroconvulsive therapy (ECT) on four treatment-resistant depressive patients (three males and one female, 55.0 +/- 17.8 years of age). The ethical committee of the National Center of Neurology and Psychiatry approved this study and written informed consent was obtained from all of the patients. ECT was administered bilaterally twice a week using atropine, propofol, and succinylcholine (or vecronium) as anesthetic medications. The Hamilton rating scale for depression scores decreased from 30.5 +/- 11.0 to 14.8 +/- 11.5 following a course of 5-12 treatments. Memory and cognitive functioning, evaluated using the Mini-Mental State Examination, Short-Memory Questionnaire and so on, had not changed a week after the last ECT. These results further support the view that brief-pulse ECT is efficacious and safe for the treatment of depression. PMID- 11062866 TI - Large scale sonochemical processing: aspiration and actuality. AB - It has been recognised for many years that power ultrasound has great potential in a wide variety of processes in the chemical and allied industries. Some of these processes have been known for many years and continue to flourish as major commercial applications, e.g. plastic welding and cleaning. Others, like ultrasonic drilling, while showing great potential have not been widely exploited to date. The potential for the industrial use of power ultrasound is enormous, and yet industry seems somewhat reluctant to adopt it. In this article the existing uses of power ultrasound in processing are reviewed and the potentials are explored. PMID- 11062867 TI - Asymmetric sonochemical reactions enantioselective hydrogenation of alpha ketoesters over platinum catalysts. AB - The sonochemical enantioselective hydrogenation of different alpha-ketoesters to the corresponding hydroxy derivatives over cinchona modified Pt catalysts is described. Ultrasonic irradiation was found to be beneficial in improving the optical yields. Besides studying the reaction rates and enantioselectivities, the effect of ultrasonics on the catalyst-modifier system and the scale-up of the process will also be considered. PMID- 11062868 TI - Ultrasound in carbohydrate chemistry: sonophysical glucose oligomerisation and sonocatalysed sucrose oxidation. AB - Both aspects of the interest in using ultrasound are illustrated by our results in the field of carbohydrate chemistry. The course of the heterogeneous reaction of glucose with hydrophobic alcohols in acidic medium is directed towards the oligomerisation of glucose because of the wetting of the glucose suspension due to the efficiency of the sonophysical mixing. On the other hand, a sonocatalysis is observed during the course of the oxidation of primary hydroxyl groups in homogeneous aqueous medium by the NaOCl/TEMPO system. PMID- 11062869 TI - Ultrasound in organic electrosynthesis. AB - Mechanical effects induced by ultrasonication can be very helpful for the activation of electrochemical reactions. The continuous cleaning of the electrodes by ultrasound irradiation of the electrochemical cell or the enhancement of mass transfer at the electrodes are examples of such activation. Finally, ultrasonication can play an important part for the orientation of reactions whose selectivities are very sensitive to stirring. Two very different examples have been chosen to illustrate these phenomena: the indirect electrooxidation of di-ketone-L-sorbose into the corresponding ketogulonic acid and the direct electroreduction of acetophenone into pinacol. PMID- 11062870 TI - Phase-transfer catalysis and ultrasonic waves. I. Cannizzaro reaction. AB - The aim of this work is to study the effect of an ultrasonic wave on the Cannizzaro reaction catalyzed by a phase-transfer catalyst. The reaction of benzaldehyde with potassium hydroxide was chosen as the reference reaction. The kinetics of the reaction was followed by the amount of benzoic acid which is well characterized and easily isolatable. Investigations were made on variables such as the kind of aldehyde, the phase-transfer catalyst, the temperature and the frequency of ultrasonic wave. As the phase-transfer catalyst depends strongly on mass transfer between two phases, it is well understood that ultrasonic waves have a greater efficiency of interface mixing than conventional agitation. The results showed that an ultrasonic wave of 20 kHz dramatically accelerates on the reaction. PMID- 11062871 TI - Ultrasonics in chemoselective heterogeneous metal catalysis sonochemical hydrogenation of unsaturated carbonyl compounds over platinum catalysts. AB - The hydrogenation of unsaturated aldehydes was studied over ultrasonically pretreated silica supported platinum catalyst. In the hydrogenation of cinnamaldehyde a remarkable reaction rate and cinnamyl alcohol selectivity increase was observed. Using other unsaturated aldehydes the reaction rate and unsaturated alcohol selectivity increased moderately. PMID- 11062872 TI - Volatile metal beta-diketonates--new precursors for the sonochemical synthesis of nanosized materials--sonolysis of thorium(IV) beta-diketonates. AB - Ultrasonic irradiation (22 kHz, Ar atmosphere) of Th(IV) beta-diketonates Th(HFAA)4 and Th(DBM)4, where HFAA and DBM are hexafluoroacetylacetone and dibenzoylmethane respectively, causes them to decompose in hexadecane solutions, forming solid thorium compounds. The first-order rate constants for Th(IV) beta diketonate degradation were found to be (9.3 +/- 0.8) x 10(-3) for Th(HFAA)4 and (3.8 +/- 0.4) x 10(-3) min-1 for Th(DBM)4, (T = 92 degrees C, I = 3 W cm-2). The rate of the sonochemical reaction increased with the rising beta-diketonate volatility and decreased with the rising hydrocarbon solvent vapor pressure. Solid sonication products consisted of a mixture of thorium carbide ThC2 and Th(IV) beta-diketonate partial degradation products. The average ThC2 particle size was estimated to be about 2 nm. ThC2 formation was attributed to the high temperature reaction occurring within the cavitating bubble. The thorium beta diketonate partial degradation products formed in the liquid reaction zones surrounding the cavitating bubbles. PMID- 11062873 TI - A comparative study between classical stirred and ultrasonically-assisted dead end ultrafiltration. AB - The aim of this work was to determine mass transfer coefficients in the cases of ultrasonically-assisted and classical stirred dead-end ultrafiltration. A comparative study was then performed, and mass transfer coefficients obtained under ultrasonic conditions are described by an empirical model. This correlation results from an analogy with what is observed using a stirred cell and involves the ultrasonic power as the main parameter. The hydrodynamics are assumed to depend on the intensity of the ultrasound effects illustrated by the agitation arising within the cell. This agitation is due to convective currents as well as physical effects due to cavitation. The concentration polarization phenomenon is therefore affected by this action of ultrasonic waves. PMID- 11062874 TI - Emulsification processes: on-line study by multiple light scattering measurements. AB - The use of ultrasound in various processes of the chemical industry has been a subject of research and development for many years. As regards in emulsification, apart from formulation variables, power is the most important parameter. Efficiency of emulsification processes may be followed and evaluated by measuring particle size distribution, which mainly governs the kinetic stability of such dispersions. Unfortunately, this kind of measurement must be performed at high dilution (low volume fraction of dispersed phase). The present work is devoted to the on-line study of ultrasound emulsification by means of a newly developed apparatus based on multiple light scattering, which allows us to determine average droplet diameter and its variations directly on concentrated media. The model system was an oil (kerosene)-in-water emulsion stabilized by a polyethoxylated sorbitan monostearate. PMID- 11062875 TI - Suppression of slag foaming by a sound wave. AB - The aim of this work was to study the effects of sound frequency, sound intensity and viscosity of slag on the slag foaming rate and the steady-state foam height. Experiments were carried out using two slags (BaO-B2O3) melted at a temperature of 1223 or 1273 K, as well as water-glycerin solutions at room temperature. Low frequency sound waves (< 1.3 kHz) are found to be more effective in the slag foaming suppression than high frequency waves (1.3-12 kHz). The steady-state foam height decreases abruptly when the sound pressure reaches a threshold value that depends on sound frequency and liquid viscosity. The results can be explained in terms of enhancing the rates of liquid drainage and film rupture induced by sound. PMID- 11062876 TI - Double-structured ultrasonic high frequency reactor using an optimised slant bottom. AB - The present work has been carried out in order to design a new type of ultrasonic reactor consisting of a double-structured tank. The inner working compartment is built with a slant bottom to allow a better ultrasonic transmission. This paper reports the effect of the inclination angle on acoustic efficiency for several different plates, e.g. two plates made of glass (2 mm and 3 mm thickness) and one made of PVC (3 mm thickness). The acoustic efficiency was determined as the ratio of the signal measured by a hydrophone in the presence of the plate to that signal in the absence of the plate. Having optimised the system, the ultrasonic powers in the inner and the outer compartments of the slant bottom reactor were determined by hydrogen peroxide dosimetry. PMID- 11062877 TI - Velocity study in an ultrasonic reactor. AB - In order to determine the parameters required to describe and to optimize sonochemical reactors, we have investigated the water flow inside such a reactor. With this aim, the experimental velocity field has been measured by tomography laser. The influence of certain parameters such as the electric power, the water height and the fluid viscosity has been evaluated. At the same time, the water movement has been studied theoretically using Nyborg's model. We have tried to improve this model by considering a three-dimensional velocity. PMID- 11062878 TI - Cavitation activity stimulation by low frequency field pulses. AB - The influence of a short-time action of a low-frequency ultrasound on the sonoluminescence generation by a high frequency pulsed field has been studied. This action remarkably lowers the cavitation thresholds and increases the sonoluminescence intensity. The stimulating effect of the low-frequency field action depends on its duration and on the intensities of both fields. Possible mechanisms of this effect are discussed. PMID- 11062879 TI - Numerical simulation of cavitation bubble dynamics induced by ultrasound waves in a high frequency reactor. AB - The use of high frequency ultrasound in chemical systems is of major interest to optimize chemical procedures. Characterization of an open air 477 kHz ultrasound reactor shows that, because of the collapse of transient cavitation bubbles and pulsation of stable cavitation bubbles, chemical reactions are enhanced. Numerical modelling is undertaken to determine the spatio-temporal evolution of cavitation bubbles. The calculus of the emergence of cavitation bubbles due to the acoustic driving (by taking into account interactions between the sound field and bubbles' distribution) gives a cartography of bubbles' emergence within the reactor. Computation of their motion induced by the pressure gradients occurring in the reactor show that they migrate to the pressure nodes. Computed bubbles levitation sites gives a cartography of the chemical activity of ultrasound. Modelling of stable cavitation bubbles' motion induced by the motion of the liquid gives some insight on degassing phenomena. PMID- 11062880 TI - High-speed video microscopy and computer enhanced imagery in the pursuit of bubble dynamics. AB - The equipment and method for studying transient bubble dynamics are described in simple sonochemical reactors and presented using still frames from high-speed video microscopy (500 fps). Effects on aeration bubbles (mean size 1-3 mm diameter) and the cavitation induced species (< 0.5 mm diameter) are studied. The images are computer enhanced to improve interpretation of such features as the maximum ellipsoidal distortion at the nodal sound plane and spherical shape regain with due consideration of energy involved and expansion effects at the nodal sound plane. Also immersion depth/pressure effects, as the bubbles transcend the sound field column, in the cylindrical reactor, are recorded for evaluation of nodal and antinodal sound wave effects. Positions of the nodal and antinodal regions are marked using a novel tungsten halogen bulb technique and verified using the sonoelectroluminescent approach with the classical luminol/hydrogen peroxide chemistry which is enhanced under the sound field conditions. PMID- 11062881 TI - Scale-up of sonic energy for a major industrial application: ash conditioning for power utilities that use fluidized bed combustion. AB - The paper describes a new, powerful, patented piece of sonic equipment that operates in the low sonic frequency range, at 104 Hz, with nominal power of 75 kW. It has been tested for both physical and chemical sonic effects. A large scale industrial application is described--the conditioning and pacification of ash from utility coal combustion by fluidized bed combustion. A variety of additional applications is suggested, most of which have been briefly tested. PMID- 11062882 TI - Dye effluent decolourisation using ultrasonically assisted electro-oxidation. AB - Solutions of the acidic dye, Sandolan Yellow, were subjected to sonolysis, electrolysis and sonoelectrolysis. Decolourisation did not take place using ultrasound alone but was achieved using an electro-oxidation process. The rate of electro-chemical decolourisation in the absence of ultrasound was dependent on the type of electrode used, electrolyte concentration, reaction temperature and the current density. Electro-oxidation of Sandolan Yellow using platinum electrodes was enhanced using ultrasound when carried out in a semi-sealed cell, which minimised the effects of ultrasonic degassing. PMID- 11062883 TI - The application of power ultrasound to the surface cleaning of silica and heavy mineral sands. AB - Power ultrasound may be used in the processing of minerals to clean their surfaces of oxidation products and fine coatings, mainly through the large, but very localised, forces produced by cavitation. Results of the application of power ultrasound to remove iron-rich coatings from the surfaces of silica sand used in glass making and to improve the electrostatic separation of mineral sand concentrates through lowering the resistivity of the conducting minerals (ilmenite and rutile) are presented. Parameters affecting ultrasonic cleaning, such as input power and levels of reagent addition, are discussed. In particular, we present data showing the relationship between power input and the particle size of surface coatings removed. This can be explained by the Derjaguin approximation for the energy of interaction between a sphere and a flat surface. PMID- 11062884 TI - Isotope effect in hydrogen peroxide formation during H2O and D2O sonication. AB - The kinetics of hydrogen peroxide formation have been studied during H2O and D2O sonication in the presence of argon and oxygen (f = 22 kHz, I = 3.0 W cm-2, Pac = 0.52 W ml-1, V = 20 ml, T = 20 degrees C). It was found that the sonochemical reaction rate W has a zero order with respect to hydrogen peroxide (H2O, D2O or DHO2) concentration. In argon atmosphere the kinetic isotope effect was found to be equal to alpha = WH2O/WD2O = 2.2 +/- 0.3. The alpha value decreases in H2O-D2O mixtures with increasing H2O concentration. In oxygen atmosphere the isotope effect is not observed (alpha = 1.05 +/- 0.10). It is assumed that the revealed isotope effect is related to the mechanism of water sonolysis including the H2O Ar* and D2O-Ar* energy transition, where Ar* is an argon atom in an excited state, in nonequilibrium plasma generated by the shock-wave. PMID- 11062885 TI - Sonochemical and photochemical oxidation of organic matter. AB - Recent developments in sonochemistry have led us to study its use to treat water and wastewater. The effects of ultrasound wave in hydrophilic chemical oxidations are mainly due to hydroxyl radical production during the cavitation-induced water decomposition. Currently, the sonochemical destruction of aromatic compounds in water solution is obtained with low rates. The aim of this work is to evaluate the efficiency of the sonochemical effect in conjunction with a photochemical irradiation. Taking phenol as an example, the combined action of sonochemistry and photochemistry has been considered in a 'sonuv' reactor. An important enhancement of the degradation rate of phenol has been observed. It may be the result of three different oxidative processes: direct photochemical action, high frequency sonochemistry and reaction with ozone (produced by UV irradiation of air). The process has been successfully tested to lower the chemical oxygen demand of a municipal wastewater. PMID- 11062886 TI - Sonolytic degradation of hazardous organic compounds in aqueous solution. AB - Benzene, chlorobenzene, 1,2-, 1,3-, 1,4-dichlorobenzene, biphenyl, and polychlorinated biphenyls such as 2-, 4-chlorobiphenyl and 2,2'-dichlorobiphenyl in aqueous solutions have been subjected to sonolysis with 200 kHz ultrasound at an intensity of 6 W cm-2 under an argon atmosphere. 80-90% of initial amount of these compounds were degraded by 30-60 min of sonication when the initial concentrations were 10-100 mumol l-1. The degradation rate of these compounds increased with increase in their vapor pressures. In all cases of sonolysis of chlorinated organic compounds, an appreciable amount of liberated chloride ion was observed. PMID- 11062888 TI - [53rd annual meeting of the Japanese Association for Thoracic Surgery. Beppu city, Japan. October 25-27, 2000. Abstracts]. PMID- 11062887 TI - Effects of ultrasound energy on degradation of cellulose material. AB - The rationale for selection of waste cellulose source and method for its degradation, such as ultrasound, aeration and coupled energy, is examined. Consideration is given to the availability of waste material for the conservation of global resources, pollution effects from energy forms and efficiency of energy transfer. Availability of the sources and possible ways of converting them to fuels, processes involved in its production and the possible effects on the environment are discussed. Manufactured cellulose and waste paper are used as the source for these experiments and the rationale behind their use in the environment is analysed. An ultrasound reactor that operates at 80 W and 38 kHz was used in breaking down the samples to produce glucose and other chemical species. One of the routes being explored is the further conversion of these molecules into fuel (alcohol). PMID- 11062889 TI - [The 40th annual meeting of the Japanese Society of Nuclear Medicine. Kobe, Japan. November 1-3, 2000. Abstracts]. PMID- 11062890 TI - [The 28th annual meeting of the Japanese Society of Clinical Immunology. Tokyo, Japan. September 28-30, 2000. Abstracts]. PMID- 11062892 TI - [Regional meeting of the Japanese Society of Otolaryngology. Japan. 2000. Abstracts]. PMID- 11062891 TI - [The 36th autumn meeting of the Japan Radiological Society. Nagasaki, Japan. September 25-27, 2000. Abstracts]. PMID- 11062893 TI - [The 73rd Congress of the Japanese Biochemical Society. Yokohama, Japan. October 11-14, 2000. Abstracts]. PMID- 11062894 TI - [Classification and diagnosis of diabetes mellitus]. PMID- 11062895 TI - [Genetic factors and life styles related to diabetes mellitus--with special reference to multiple factor inheritance]. PMID- 11062896 TI - [Physiopathology of diabetes mellitus--insulin secretory dysfunction or resistance? 1) Secretory dysfunction]. PMID- 11062897 TI - [Physiopathology of diabetes mellitus--insulin secretory dysfunction or resistance? 2) Insulin resistance]. PMID- 11062898 TI - [Approaches to diabetes therapy. 1. Diet and exercise therapy]. PMID- 11062899 TI - [Approaches in diabetes therapy. 2. Selection and use of oral antidiabetics. 1) Drugs enhancing insulin secretion]. PMID- 11062900 TI - [Approach to diabetes therapy. 2. Selection and the use of oral antidiabetics. 2) Agents to correct insulin resistance]. PMID- 11062901 TI - [Approach to diabetic therapy. 2. Selection and the use of oral antidiabetics. 3) Biguanides and alpha-glucosidase inhibitors]. PMID- 11062902 TI - [Approach to diabetes therapy. 3. Insulin preparations--their development and progress in diabetes therapy]. PMID- 11062903 TI - [Approach to diabetes therapy. 4. Control standard with a consideration given to complications and age of patients]. PMID- 11062904 TI - [Diagnosis, treatment, and management of diabetic complications. 1. Foot care]. PMID- 11062905 TI - [Diagnosis, treatment, and management of diabetic complications. 2. Neurologic disorders]. PMID- 11062906 TI - [Diagnosis, treatment, and management of diabetic complications. 3. Non-surgical treatment of diabetic retinopathy]. PMID- 11062907 TI - [Diagnosis, treatment, and management of diabetic complications. 4. Nephropathies]. PMID- 11062908 TI - [Diagnosis, treatment, and management of diabetic complications. 5. Hypertension]. PMID- 11062909 TI - [Diabetic emergency management. 1. Acute complications]. PMID- 11062910 TI - [Response to diabetic emergencies. 2. Sick days]. PMID- 11062911 TI - [Emergency care of diabetics. 3. Hypoglycemia and unstable metabolic state (including self-determination of the blood glucose level)]. PMID- 11062912 TI - [Care of diabetic patients. 1. The importance of cooperation among medical specialties]. PMID- 11062913 TI - [Care of diabetic patients. 2. The importance of patient education]. PMID- 11062914 TI - [Care of diabetic patients. 3. The role and use of certified diabetes educators]. PMID- 11062915 TI - [Current trend in the care of diabetic patients: discussion]. PMID- 11062916 TI - [Intrafamilial epidemic of meningococcal meningitis]. PMID- 11062917 TI - [Aortitis syndrome with nasal angiitis and positive P-ANCA]. PMID- 11062918 TI - [Hypothyroidism with severe cerebellar atrophy and ataxia]. PMID- 11062919 TI - [Efficacy of the extra-corporeal lung assist (ECLA) on the alveolar hemorrhage in a case with microscopic polyarteritis nodosa]. PMID- 11062920 TI - [Quadruple malignant neoplasms composed of gastric carcinoid, prostatic carcinoma, malignant lymphoma, and malignant mesothelioma in a short-term]. PMID- 11062921 TI - [Pulmonary thromboembolism and its molecular behavior]. PMID- 11062922 TI - [Progress in echocardiography]. PMID- 11062923 TI - Exposure potentials during cleaning, overhauling and repairing of aircraft lavatory tanks and hardware. PMID- 11062924 TI - Sulfur dioxide exposure in an electroplating establishment. PMID- 11062925 TI - Task-based exposure assessment of hazards associated with new residential construction. PMID- 11062926 TI - Control of drywall sanding dust exposures. National Institute for Occupational Safety and Health. PMID- 11062927 TI - Indoor air quality in a middle school, Part I: Use of CO2 as a tracer for effective ventilation. AB - The overall objective of the study was to evaluate the indoor air quality at a middle school with an emphasis on characterizing baseline conditions. The focus of this article is on the relationship between occupancy and measured concentrations of carbon dioxide, and an evaluation of the use of carbon dioxide as a tracer for ventilation in the school. The school was characterized as having no health complaints, good maintenance schedules, no carpeting within the classrooms or hallways, and no significant remodeling, and its officials had agreed to allow the sampling to take place during school hours. Monitoring followed the guidelines recommended in the "Preliminary Draft: Conceptual Standardized EPA Protocol For Characterizing Indoor Air Quality in School Buildings." Four indoor locations including the cafeteria, a science classroom, an art classroom, and the lobby outside the main office, and one outdoor location were sampled for various environmental comfort and pollutant parameters for one week in February 1997. A consistent relationship between hourly occupancy and corresponding carbon dioxide concentrations was seen. Carbon dioxide concentrations in the cafeteria, art room, and lobby were within specified American Society of Heating, Refrigerating, and Air-Conditioning Engineers (ASHRAE) guidelines for comfort (< 1000 ppm). The science room had the highest concentrations (frequently exceeding 1000 ppm) due to high occupancy and non functioning unit ventilators. Measured ventilation rates were within specified ASHRAE guidelines for the art room, cafeteria, and lobby. The science room, which relied on natural ventilation only, was not able to meet the ASHRAE guideline on one of the three days studied. The use of a completely mixed space, one compartment mass balance model with estimated CO2 generation rates and measured CO2 concentrations is shown to be a useful method for evaluating ventilation. Modeled effective ventilation, air changes per hour, and mixing factors reflected measured carbon dioxide concentrations and measured ventilation in each room. Mechanical ventilation afforded better mixing than natural ventilation. This study demonstrates the usefulness of collecting indoor CO2 and occupancy data when carrying out indoor air quality evaluations in schools. PMID- 11062928 TI - Indoor air quality in a middle school, Part II: Development of emission factors for particulate matter and bioaerosols. AB - A middle school (grades 6 to 8) in a residential section of Springfield, Illinois, with no known air quality problems, was selected for a baseline indoor air quality survey. The study was designed to measure and evaluate air quality at the middle school with the objective of providing a benchmark for comparisons with measurements in schools with potential air quality problems. The focus of this article is on the development of emission factors for particulate matter and bioaerosols. The school was characterized as having no health complaints and good maintenance schedules. Four indoor locations including the cafeteria, a science classroom, an art classroom, the lobby outside the main office, and one outdoor location were sampled for various environmental comfort and pollutant parameters for one week in February 1997. Integrated samples (eight-hour sampling time) for respirable and total particulate matter, and short-term measurements (two-minute samples, three times per day) for bioaerosols were collected on three consecutive days at each of the sampling sites. Continuous measurements of carbon dioxide were logged at all locations for five days. Continuous measurements of respirable particulate matter were also collected in the lobby area. A linear relationship between occupancy and corresponding carbon dioxide and particle concentrations was seen. A completely mixed space, one compartment mass balance model with estimated CO2 generation rates and actual CO2 and particulate matter concentrations was used to model ventilation and pollutant emission rates. Emission factors for occupancy were represented by the slope of emission rate versus occupancy scatter plots. The following particle and bioaerosol emission factors were derived from the indoor measurements: total particles: 1.28 mg/hr/person-hr; respirable particles: 0.154 g/hr/person-hr; total fungi: 167 CFU/hr/person-min; thermophilic fungi: 35.8 CFU/hr/person-min; mesophilic fungi: 119 CFU/hr/person-min; total bacteria: 227 CFU/hr/person-min; gram-negative bacteria: 69.5 CFU/hr/person-min; gram-positive bacteria: 191 CFU/hr/person-min; Aspergillus: 17.0 CFU/hr/person-min; Penicillium: 161 CFU/hr/person-min; and yeasts: 16.4 CFU/hr/person-min. PMID- 11062929 TI - Pollution prevention and the work environment: the Massachusetts experience. AB - In 1989, Massachusetts enacted the Toxics Use Reduction Act. The Act defined toxics use reduction, also referred to as pollution prevention, as "in-plant changes in production processes or raw materials that reduce, avoid, or eliminate the use of toxic or hazardous substances or generation of hazardous by-product per unit of product ... without shifting risks between workers, consumers or parts of the environment." The investigators sought to understand to what extent worker health and safety concerns have been integrated into toxics use reduction activities and how these activities have affected the work environment. The authors reviewed 35 published case studies of toxics use reduction in Massachusetts companies and interviewed key personnel including the staff of the Massachusetts Office of Technical Assistance for Toxics Use Reduction. Overall, between 1990 and 1997, Massachusetts companies decreased their use of toxic chemicals by 24 percent and decreased their volume of toxic by-product by 41 percent. In almost 50 percent of the cases analyzed, improved worker health and safety was cited as a benefit of the toxic use reduction projects. Solvents were eliminated or reduced in 63 percent of the cases. Forty-six percent of the companies profiled introduced water-based chemicals in place of more volatile ones; and acids and caustics were reduced or eliminated in 20 percent of the cases. The investigators concluded that toxics use reduction activities have resulted in improvements to the work environment, but that such improvements were rarely a direct concern of these efforts, thus creating the potential for new negative worker health and safety impacts and missed opportunities to coordinate environmental and worker health and safety improvements. The authors recommend that technical assistance agencies and companies better integrate worker health and safety issues and pollution prevention activities. PMID- 11062930 TI - Utilization of health and safety consulting services of the Ohio Bureau of Workers' Compensation by small businesses. AB - The Division of Safety and Hygiene (DSH) of the Ohio Bureau of Workers' Compensation (OBWC) provides health and safety consulting services to all businesses in Ohio insured by OBWC. It has been conceptualized that small companies in Ohio do not use health and safety consultative services to the same extent as non-small companies. To determine the validity of this belief, this study was conducted to analyze the records of the DSH for the years 1991-1995 in the Division's Southeast Region which contained a total of 281,041 insured companies. Of the 5977 companies utilizing the health and safety services of DSH and recording their number of employees, 65.3 percent were "small," defined as any company with less than 20 employees. Specific services used by businesses included industrial hygiene (small, 3.4% vs. non-small, 6.4%), ergonomics (0.7% vs. 2.8%), engineering such as ventilation or machine guarding (0.6% vs. 1.7%), construction safety (26.7% vs. 3.9%), and general safety such as implementing a safety program (31.4% vs. 22.4%). In conclusion, industrial hygiene, ergonomics, and engineering service utilization were significantly lower for the small businesses than for the non-small businesses, whereas safety service utilization was significantly higher for the small business. Improvement in the system of record-keeping and education/encouragement in the deployment of services are recommended. PMID- 11062931 TI - Regulated workplace ketones and their interference in the PFBHA method for aldehydes. AB - Ketones are the major positive interferences for an aldehyde dynamic air sampler that consists of 200-mg 20 percent (w/w) O-(2,3,4,5,6 pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA) on Tenax TA contained in a Pyrex tube 7-mm OD, 5-mm ID, and 70-mm in length, that utilizes a personal battery-powered pump at 10-50 mL/min. The ketone O-oxime derivatives were synthesized to allow absolute quantitation of O-oximes formed during sampling. Wet spiking allowed ketone recoveries to be found. Ketone vapors of known concentrations were generated statically in Tedlar gas bags. The O-oximes were desorbed with hexane, and an aliquot injected for gas chromatographic analysis on a nonpolar capillary column with mass spectrometric or electron capture detection. Gas phase recoveries up to 200 ppm-hour loadings exceeded 75 percent at 25 degrees C for chloroacetone, cyclohexanone, diacetone alcohol, diethyl ketone, dipropyl ketone, ethyl butyl ketone, methyl amyl ketone, methyl butyl ketone, 2-methylcyclohexanone, methyl ethyl ketone, methyl isobutyl ketone, methyl isopropyl ketone, and methyl propyl ketone. The recoveries for acetophenone, 2-chloroacetophenone, and ethyl amyl ketone were lower than 75 percent, and were caused by steric hindrance. Sampling for both aldehydes and ketones is recommended at 10 mL/min for TLV concentrations. PMID- 11062932 TI - The effect of thermal loading on laboratory fume hood performance. AB - A modified version of the ANSI/ASHRAE 110-1995 Method of Testing Performance of Laboratory Fume Hoods was used to evaluate the relationship between thermal loading in a laboratory fume hood and subsequent tracer gas leakage. Three types of laboratory burners were used, alone and in combination, to thermally challenge the hood. Heat output from burners was measured in BTU/hr, which was based on the fuel heat capacity and flow rate. Hood leakage was measured between 2824 and 69,342 BTU/hr. Sulfur hexafluoride (SF6) was released at 23.5 LPM for each level of thermal loading. Duct temperature was also measured during the heating process. Results indicate a linear relationship for both BTU/hr vs. hood leakage and duct temperature vs. hood leakage. Under these test conditions, each increase of 10,000 BTU/hr resulted in an additional 4 ppm SF6 in the manikin's breathing zone (r2 = 0.68). An additional 3.1 ppm SF6 was measured for every 25 degrees F increase in duct temperature (r2 = 0.60). Both BTU/hr and duct temperature models showed p < 0.001. For these tests, BTU/hr was a better predictor of hood leakage than duct temperature. The results of this study indicate that heat output may compromise fume hood performance. This finding is consistent with those of previous studies. PMID- 11062933 TI - Tooth loss and implant replacement. AB - Osseointegrated dental implants are increasingly used to replace missing teeth in a variety of situations ranging from the missing single tooth to complete edentulism. The implant possibility must be carefully considered because treatment involves extended time frames, considerable expense and is not without risk. Accordingly, treatment-planning decisions should have an evidence-based strategy with appropriate risk assessment. Implant systems need to be adequately tested before they are released for general use and success rates should be assessed from peer review scientific publication data and not commercial promotional literature. It is the responsibility of the dentist to ensure the patient is educated so an informed decision can be made on difficult treatment alternatives. The clinical decision making process must respect the issues to assure quality of care and reduction of liability for negligent care. Today, the three-unit fixed bridge can no longer be considered as the standard of care for restoration of a single missing tooth. The evidence has accumulated that the single tooth implant supported replacement is more conservative, more cost effective and more predictable with respect to long-term outcome in uncomplicated cases. PMID- 11062934 TI - Current concepts on functional appliances and mandibular growth stimulation. AB - The mode of action of functional appliances, particularly in relation to stimulating mandibular growth, is a controversial subject. Many of the reports concerning growth effects of functional appliances have been characterized by poor methodology. In assessing functional appliances, results from prospective randomized clinical trials should be given prominence. On the basis of available evidence, it cannot be concluded that functional appliances are effective in stimulating and increasing mandibular growth in the long term. Although favourable growth changes have been reported following phase 1 therapy, they are generally not substantial and long term stability appears to be poor. PMID- 11062935 TI - Antibiotic prescribing practices by South Australian general dental practitioners. AB - The prescribing habits of a randomly selected approximately 10 per cent sample of South Australian general dental practitioners were obtained by postal questionnaire. Sixty-eight (61 per cent) usable replies were received and analysed. Generally, there was an appropriate level of knowledge of antibiotic prescription. However, there was a tendency toward over-prescription and a demonstrated lack of knowledge of the incidence of adverse reactions, development of multiresistant strains and prophylaxis against bacterial endocarditis. All of these areas are real challenges to the profession, whether in an overall global community health sense or in a highly individualized clinical or medico-legal sense. These issues are discussed and the profession is urged to reconsider and re-educate itself on these challenges. PMID- 11062936 TI - Potential of 4 per cent silver fluoride to induce fluorosis in rats: clinical implications. AB - The Health Department in Western Australia uses a 40 per cent silver fluoride (AgF) solution for prevention and treatment of dental caries in children. Analysis of this solution has revealed high fluoride concentrations (75,000 120,000 mg/L), raising concerns of potential toxicity and prompting investigation of clinical protocols utilizing low-strength AgF in an animal model. A single topical application of 4 per cent AgF solution to Sprague-Dawley rats resulted in moderate to severe localized fluorosis in 24 per cent of animals. In a second experiment, caries was induced in rats aged 19 days; six weeks later, between one four carious molar teeth from each rat were treated with 4 per cent AgF (atraumatic technique). A generalized form of fluorosis developed in the continually growing incisors of less than 10 per cent of animals which had one or two carious teeth treated, and in 70-90 per cent of rats which received AgF to either three or four carious teeth. These results confirm the potential of a 4 per cent AgF solution to induce fluorosis and support previous recommendations that AgF at its empirical concentration of 40 per cent should be withdrawn from clinical use. PMID- 11062937 TI - A survey of the distribution and types of full crowns prescribed in Melbourne, Australia. AB - The full crowns made by 11 commercial dental laboratories in Melbourne, Australia, were surveyed over a two-month period. The survey was designed to find which teeth were crowned and what types of full crowns were prescribed. Maxillary central incisors were found to be the most frequently crowned teeth (15.6 per cent) and porcelain-fused-to-metal crowns were the most commonly prescribed type of crown (82.9 per cent). This study may form a baseline with which other Australian studies may be compared and contrasted. PMID- 11062938 TI - In vitro fluoride release from aesthetic restorative materials following recharging with APF gel. AB - In this in vitro study, the fluoride ion release from three more-viscous conventional glass ionomer cements (GICs)--ChemFlex, Fuji IX GP, Ionofil Molar- four polyacid-modified resin composites (compomers)--Dyract AP, Compoglass F, Freedom, F2000--and a hybrid resin composite--Ariston pHc--were compared. The amounts of fluoride release from 3 x 2.7 mm specimens were measured over six weeks using a fluoride ion-selective electrode. After six weeks, the specimens were recharged with 2 ml of 1.23 per cent acidulated phosphate fluoride (APF) gel for four minutes. The recharged specimens were then assessed for the amounts of fluoride release over another six weeks. Statistical analyses were performed using one-way and repeated measures ANOVA. The GICs and Compoglass F showed significantly higher initial fluoride release rates during the first two days (p < 0.05). After the first two days, fluoride release rates from all materials dropped quickly and became essentially stabilized within three-five weeks, in an exponential mode. The recharging of the specimens with APF gel caused a brief, but significant, increase in fluoride release for all materials (p < 0.05), before decreasing to previous comparable rates. Compoglass F released relatively more fluoride, and Ariston pHc relatively less, after APF gel application than before. The lowest total amounts over the study were released by Dyract AP and Freedom. The newer compomers, as well as the newer more-viscous GICs, appear to act as fluoride reservoirs to varying extents. PMID- 11062939 TI - Oral health and hospitalization in Western Australian children. AB - Over the past 20 years, the prevalence of dental disease in Western Australian children has diminished. The causes of this significant improvement in health are associated with better care models, water fluoridation and changes in lifestyle. In this study, the authors examine the reasons for hospitalization for oral health conditions in Western Australia for the calendar year 1995 using the Health Department of WA database. A total of 3,754 episodes of care (4,395 bed days) was recorded for dental conditions. Dental caries resulted in the fifth and sixth highest number of episodes of hospitalization in preschool (1-4 years) and primary-school age (5-12 years) children respectively. Abnormal tooth eruption resulted in the highest number of episodes of hospitalization in high-school age (13-17 years) children. From the age-stratified rates of hospitalization (per 1000), non-Aboriginal children were more than twice as likely to enter hospital for dental related conditions. The primary cause of this is the 15 times higher rate of hospitalization for high-school age non-Aboriginal children which clearly reflects the greater use of services for impacted third molars by the metropolitan non-Aboriginal community. Examination of the distribution by health service region revealed the hospitalization rate was significantly less than the state average for the Kimberley, Pilbara, Northern Goldfields and Wanneroo regions. These data reflect the paucity of oral health care available to residents of these regions, particularly the northwest, and does not reflect a diminished burden of disease. Similarly, the rate of hospitalization for Aboriginal children reflects population and service delivery differences particularly in regional and remote WA. These data highlight the need to develop new strategies in oral health care to target 'at risk' groups in the community, particularly new parents of young children. The preventive measures associated with good oral health in children are clearly aligned with those for good general health and can be integrated into existing health messages. PMID- 11062940 TI - Are dental radiographs safe? AB - Dental patients are often aware that radiation has the potential to harm them but they do not usually understand how or why and what potential harmful effects may arise from dental radiographs. The potential for undesirable effects must be balanced against the benefits obtained from radiographs. Dentists should address the concerns of patients who question the need for radiographs and allow them to make an informed decision. Data are available that relate radiation exposure levels from medical and dental radiographs to normal background exposure levels and allow comparisons with everyday risks in life. Recognized radiation authorities publish guidelines to help dentists with their use of radiographs, although, due to the time lag associated with testing and the publication of results, some of the published data may not always be entirely relevant to currently used X-ray machines and techniques. Dentists also have professional obligations not only to limit the use of radiographs to potentially beneficial situations but also to take good quality diagnostic radiographs, to limit the doses used, to use good radiation safety measures and to use modern equipment to achieve the best possible films. Radiographs must then be properly developed and viewed under appropriate conditions to gain the maximum possible diagnostic information from each exposure. PMID- 11062941 TI - Thin-section machine. PMID- 11062942 TI - Preventive dentistry. PMID- 11062943 TI - Temporomandibular joint arthroscopy. PMID- 11062944 TI - [Experience with 100 liver transplant recipients at the Rabin Medical Center and the Schneider Children's Medical Center]. AB - Liver transplantation is the treatment of choice for end-stage liver disease. During the past 8 years we performed 102 liver transplants in 84 adults and 16 children. In the adults, 9 were combined transplants: 1 a liver-pancreas transplant for type I diabetes, and 8 liver-kidney transplants. In the children, transplants included 5 whole-livers, 5 left-lateral liver segments from living related donors, 4 reduced-grafts of right or left lobes, and 2 split left-lateral segments. At a mean follow-up of 31 months (range 1-96) 70 were alive, 3 had died during surgery and 15 during the first postoperative months. Mortality was due to primary graft non-function (7), sepsis (10), intracranial hemorrhage (1), tumors (4), recurrent hepatitis B (2), biliary strictures (2) and chronic rejection (1). The 1- and 4-year survival rates were 79.5% and 69.6%, respectively. After transplantation, 10 developed biliary stricture (5 corrected by balloon dilatation) and 8 anastomotic stricture (7 corrected by surgery), and there were 2 multiple intrahepatic strictures. There was hepatic artery thrombosis in 5, including 4 children. In 3, grafts were salvaged by thrombectomy and 2 others underwent re-transplantation. In those who survived transplantation by more than 1-month, recurrent hepatitis B was seen in 6 of 17 (35%) and recurrent hepatitis C in 12 of 19 (63%). Thus, results of our first 100 liver transplants are similar to those reported by larger centers, showing that in an appropriate setting good results can be achieved by small transplant programs. PMID- 11062945 TI - [High-dose chemotherapy and autologous stem cell transplantation for refractory and relapsing Hodgkin's disease as first-line therapy-- studies at Sheba Medical Center--Tel Hashomer]. AB - High dose chemotherapy and autologous stem cell transplantation are widely used in relapsed and primary refractory Hodgkin's disease. We transplanted 42 patients with Hodgkin's disease between 1990-1998. Median follow-up was 31 months (range 1 102). 29 (69%) were transplanted after relapse and 13 (31%) were refractory to first line therapy. Median age at transplantation was 29 years (range 19-58) and 23 (55%) were males. All were treated with the BEAM protocol (carmustine, etoposide, cytarabine and melphelan). 18 who were in remission received radiotherapy following transplantation. The source of the stem cells was bone marrow in 17% and peripheral blood in 83%. At initial diagnosis: 57% had stage III-IV disease and B symptoms were present in 52%. 75% were treated with MOPP, ABVD or with related versions. Radiotherapy followed in 52%. Prior to transplantation, 45% of the relapsed group were in the advanced stage. 33% and 12% of all patients had lung and bone involvement, respectively. The complete remission rate was 86% for the 2 groups. 2 (5%) died from transplant-related complications and MDS/AML developed in 2 (5%) after transplantation. The 3-year overall survival (OS) and disease-free survival (DFS) were 68% and 60%, respectively. The 3-year OS for the relapsed group was 64% compared with 76% for the refractory group, and the 3-year DFS for the relapsed group was 60% vs. 42% for the refractory group (neither difference significant). Radiotherapy following transplantation did not have a beneficial effect on DFS. No prognostic factors for outcome of transplantation were found, most probably due to the limited number of patients and the high variability of disease characteristics. We conclude that high dose chemotherapy and autologous stem cell transplantation are effective and relatively safe for relapsed or primary refractory Hodgkin's disease. The DFS at 3 years was longer for those transplanted after relapse than those with primary refractory disease, but not significantly. Patients with primary refractory disease can be salvaged with high dose chemotherapy. PMID- 11062946 TI - [Immunohistochemical identification of testicular germ cells--preliminary report]. AB - The use of testicular spermatozoa for intracytoplasmic sperm injection introduced a new treatment modality for management of male infertility. Since testicular biopsies of non-obstructive azoospermic men are not homogenous in their histological patterns, identification with certainty of focal spermatogenesis might be difficult, particularly in those with small foci of spermatogenesis. We used an immunohistochemical marker of the male germ line, an antibody generated against RBM (RNA-binding-motif), to recognize with high precision the presence of germ cells in the biopsy. Biopsies of 30 men with azoospermia, most with non obstructive azoospermia and a few with obstruction of the vas deferens, were evaluated. Immunohistochemical staining for RBM protein contributed to the detection and accuracy of the identification of germ cells. Furthermore, this immunohistochemical technique aided the histopathologist to focus on even small foci of spermatogenesis. Absence of the protein expression confirmed the diagnosis of Sertoli-cell-only syndrome. The results indicate that expression of RBM can be a diagnostic marker for identifying the germ cells of small concentrations of spermatogenesis. This method can enhance the accuracy of histopathological evaluation of testicular biopsies that had formerly relied mainly on hematoxylin-and-eosin staining. PMID- 11062947 TI - [Penile prosthesis for erectile dysfunction--long-term follow-up]. AB - Our armamentarium for the treatment of erectile dysfunction has recently been expanded by addition of Viagra and the MUSE. However, their long-term results are still unknown. The insertion of a penile prosthesis is invasive, expensive, and irreversible, but under optimal condition provides an acceptable, definitive solution for erectile dysfunction. We evaluated our long-term results with penile prosthesis insertion (PPI). From 1987-1998, 57 patients underwent PPI in our department. Mean age was 55 years and the common causes of erectile dysfunction were atherosclerotic disease (23), radical pelvic surgery (15), and diabetes mellitus (14). Semirigid prostheses were inserted in 12 and inflatable prostheses in 45, including 42 single-component and 3 multi-component prostheses. Recently we interviewed these patients by telephone, using a standard questionnaire. Those not satisfied with the surgical results (83% of the living patients) were examined in our clinic. Mean follow-up was 53 months. In 37 (84%) the prosthesis was mechanically functional (rates after 1, 5 and 10 years were 87.8%, 80%, and 75%, respectively). In only 2 (2.5%) had serious complications led to prosthesis removal. All mechanical failures had occurred in those with inflatable prostheses after a mean of 48.5 months (range 4-113). At the time of the survey 68% were sexually active and 64% were satisfied with the surgical result. We conclude that PPI is safe treatment for erectile dysfunction. Although the rate of mechanically functioning prostheses decreases with time, modern multi-component prostheses may lead to better mechanical results. PMID- 11062948 TI - [Femoral artery pseudo-aneurysms--changes in treatment, report of 7 years]. AB - The femoral artery remains the most used peripheral site for radiological catheter access. With a greater number of both diagnostic and therapeutic procedures being performed by interventional radiologists and cardiologists, and with larger catheters being used for stenting and endovascular grafting, the incidence of iatrogenic pseudo-aneurysms reported has reached as high as 0.5-2%. Ideally, they should thrombus spontaneously. However, when this does not occur, management options include: observation, ultrasound-guided obliterative compression, direct thrombin injection, embolization, stent graft insertion, and very rarely-surgery. During a 7-year period (1992-1999) we treated 131 cases of femoral artery false aneurysms. Until 1998 ultrasound-guided compression obliteration, with a 95% success rate, was our method of choice. Since 1998, direct thrombin injection, with 100% success in 24 cases, has become our preferred method. It is pain-free, fully successful even in anticoagulated patients, and is currently our treatment of choice. PMID- 11062949 TI - [Rational use of albumin]. AB - The use of albumin has been a matter of debate since its introduction in the 1940's. Albumin is not only expensive but may also be harmful when administered inappropriately. Until recently our use of albumin was controlled by a number of authorized physicians who signed all albumin prescriptions. In August 1998, a multidisciplinary team reviewed the indications for albumin use and introduced simple guidelines for its supply and administration. As a result, the use of albumin has decreased by almost 70%. This indicates that rational use of albumin can be achieved by appropriate guidelines, without requiring administrative limitations. We believe that this conclusion holds true for other diagnostic and therapeutic procedures as well. PMID- 11062950 TI - [Marked creatine-phosphokinase elevation in myopathy after treatment with bezafibrate]. AB - Bezafibrate is a fibric acid derivative which has been widely used in the past 15 years. Recent studies have elucidated much of its mechanism of action, which mainly results in reduction of VLDL and triglyceride levels and in elevation of HDL. The drug is relatively safe and its side-effects well known, mild, and reversible. The most severe side-effect is myositis, varying from mild flu-like symptoms to rhabdomyolysis, which is extremely rare. The underlying situations most frequently associated with bezafibrate-induced myositis are renal insufficiency and concomitant treatment with certain other drugs. We describe 2 women who developed severe myositis with bezafibrate treatment. 1, aged 43, who had moderate diabetes but no renal insufficiency, was treated with metformin and warfarin, which can interact with bezafibrate and affect its metabolism. The other, aged 54, had renal insufficiency and was on home peritoneal dialysis. Her bezafibrate dose had been increased because of very high triglyceride levels. The aim of the study is to call attention to this significant side-effect of benzafibrate and to ways of preventing it. PMID- 11062951 TI - [Treatment of sputum retention by minitracheostomy]. AB - Maintenance of bronchopulmonary hygiene is mandatory for preventing complications of respiratory therapy in the hospitalized patient. Removal of secretions from the tracheobronchial tree is crucial. Conventional therapy, designed to assist in dislodging airway secretions, includes chest physical therapy, incentive spirometry, transnasal endotracheal suctioning and bronchoscopy. Minitracheostomy was first described by Matthews and Hopkinson for recurrent endotracheal suctioning in 1984. Since then there have been few papers about it, but they report good results with low morbidity. Despite this, its use is not popular in routine clinical work. We report our experience with minitracheostomy in the prevention of sputum retention. We conclude that its use is easy, safe and very effective in preventing postoperative and post-traumatic respiratory complications. PMID- 11062952 TI - [A structured course for primary care physicians on breaking bad news]. AB - Physicians are frequently required to break bad news to their patients. Previous research has shown that inconvenience, incompetence, and difficulty in dealing with patients' feelings are the main complaints expressed by physicians after such an encounter. Current educational programs dealing with breaking bad news are usually short, given in lecture format, and are inadequate in addressing essential issues such as knowledge, personal beliefs and attitudes, and previous personal experiences of physicians in such situations. In the past 8 years our Dept. of Family Medicine has implemented a course in breaking bad news that addresses these issues. A senior family practitioner and a medical social worker conduct 14 sessions of discussions and role-playing for small groups of residents and primary care physicians. The program is based on: theory dealing with methods of managing stress and crisis intervention, clarifying personal attitudes, discussions of previous personal encounters of the participants, various modalities of communication, methods of addressing patients' feelings and emotions, and coping with the emotions of the one breaking the bad news. On a 1-5 Likert scale questionnaire the course received an overall score of 4.47 (SD 0.51). Participants noted that they gained relevant communication skills for future patient encounters. A reliable examination of practitioners' competence in breaking bad news is mandatory in order to assess the efficiency of such courses. PMID- 11062953 TI - [Systemic mastocytosis]. AB - Mastocytosis has a highly variable clinical expression, and systemic mastocytosis is occasionally associated with a myeloproliferative or a myelodysplastic disorder. These patients often present without skin involvement and have a very poor prognosis. We report a 72-year-old man with this condition who had spells of flushing and dyspnea, myelofibrosis, and high serum and urine histamine levels. PMID- 11062954 TI - [Acute normovolemic hemodilution as a method of decreasing exposure to allogenic blood transfusion during surgery]. PMID- 11062955 TI - [Long-term cryopreservation of oocytes--outcome and related ethical, legal and religious issues]. PMID- 11062956 TI - [A national population-based screening program for cervical cancer in Israel, need for re-evaluation]. PMID- 11062957 TI - [Photodynamic therapy in age-related macular degeneration]. PMID- 11062958 TI - [Self-injury behavior in eating disorders]. PMID- 11062959 TI - [Arthrocentesis and lavage of the temporomandibular joint: treatment of closed lock]. PMID- 11062960 TI - [Corneal injuries during general anesthesia]. PMID- 11062961 TI - [The illness of Rabi Judah Hanasi, in the light of modern medicine]. PMID- 11062962 TI - [Treatment of phenylketonuria due to dihydropteridine reductase deficiency]. AB - Most cases of hyper-phenylalaninemia are due to deficiency of phenyl-alanine hydroxylase that converts phenyl-alanine to tyrosine. This enzymic reaction is facilitated by the co-factor tetrahydrobiopterin (BH4). A defect in the latter substrate leads to increased phenyl-alanine in 1-2 cases per million live births. Such cases are characterized by a degenerative brain process, and pronounced neurologic symptoms that cannot be prevented by a low phenyl-alanine diet alone. In 3 male newborns a deficiency of dihydropteridine reductase (DHPR) activity was diagnosed, the last of a sequence of 3 enzymes involved in the formation of BH4. Successful outcome of treatment, as well as the results of failure to diagnose and treat affected newborns, are described, with emphasis on the logistic problems involved in mass screening. PMID- 11062963 TI - [Knowledge, attitudes and skills of family physicians in Israel with regard to chronic pain management in cancer]. AB - Prolonged suffering from pain can affect quality of life to the point where patients lose desire to live. The attitude towards pain management in cancer patients has changed over the last two decades. The principles of treatment are based on prescribing analgetics while individually adapting medications and dosage according to the level of pain. Use of existing guidelines may relieve pain symptoms in 90% of patients. We evaluate knowledge, attitudes and skills of family physicians in Israel with regard to treatment of pain in cancer patients. The study included 123 family medicine residents and specialists from 3 different regions in Israel (Negev, Emek-Izrael, Jerusalem) who filled a self-reply, structured questionnaire. The response rate was 68%. The majority of physicians were residents in family medicine (55%) with work experience of 4-10 years, whose average age was 38.5 +/- 5.5 years. The majority of the study population: believe that treatment of pain in cancer patients is their responsibility (57%); do not believe that strong opioids cause addiction while it is thought that there is a moderate to high risk of addiction; feel comfortable prescribing opioids and are aware that opioids should be prescribed as needed without restriction. However, 25% will prescribe opioids "as needed" to a certain limit, and 30% will prescribe opioids only with a recommendation from the oncologist; most feel lack of knowledge in the areas of pain evaluation, opioid treatment, and managing side effects of opioid treatment and feel a need for additional knowledge of opioid use. The treatment of cancer patients and their families is an important and serious challenge facing the family physician. The teaching of the proper tools will assist the future generation of family physicians in facing this challenge. PMID- 11062964 TI - [Treatment of adults with acute lymphoblastic leukemia--results at the Sheba Medical Center, Tel-Hashomer, 1977-1988]. AB - Acute lymphoblastic leukemia (ALL) is a malignant disease whose incidence is relatively low among adults, unlike in children. Adults with ALL have a lower rate of long-term disease-free survival. During the last 20 years, a German multicenter group has shown that their protocols have achieved good results in adults ALL. We reviewed the medical records of 35 ALL patients, aged 19-63 years, whom we treated with these protocols (1988-1997). The remission rate was 94%. At a median follow-up of 46 months the 2-year overall survival was 54% and the disease-free survival was 94%. Although 2 patients died of bone marrow transplant complications, no death was directly associated with drug toxicity. The main grade 3 or 4 side effects (WHO classification) were neutropenia (91%), thrombocytopenia (71%) and anemia (71%). With there protocols we achieved high overall and disease-free survival rates, especially in comparison with other reports. Despite the high rate of severe treatment toxicity, there were no fatalities directly related to treatment. These results emphasize the need to concentrate treatment of adult ALL patients in large medical centers with expertise in the use of the complicated treatment protocols required. PMID- 11062965 TI - [Secondary bone grafting in cleft lip and palate]. AB - Results of reconstruction of residual alveolar bone defects in 52 patients operated between 1990-1998 were evaluated clinically and radiographically in a retrospective study. Ages ranged between 9-37; 30 were males. The donor site of bone grafts in all was particulate cancellous marrow from the anterior iliac crest. 32 had unilateral clefts and 20 bilateral. Total cleft sites treated was 72. Best results were achieved when bone grafting was carried out prior to the eruption of the canine tooth. The cleft space was closed and oro-nasal fistulas were eliminated in 42 (80%). Success rates in unilateral and bilateral cases were significantly different. PMID- 11062966 TI - [Reactions of patients to complementary medicine]. AB - 350 patients attending 11 large out-patient clinics completed questionnaires evaluating attitudes to, and experience with complementary medicine. 129 (36%) respondents reported using complementary medicine. 14% of them used complementary medicine for the current medical problem for which they were attending the clinic. Pain was the most common medical problem for which complementary medicine was used, followed by respiratory problems and cancer. Common therapeutic modalities used were acupuncture, homeopathy, nutrition and herbal medicine. Women, the secular as opposed to the religious, and those with higher education were more apt to use complementary medicine. No differences were found in age, national origin, length of living in Israel, and diet (vegetarian, natural foods or regular diet) between those who used complementary medicine and those who did not. No relationship was found between the use of complementary medicine and perceived poor health status, locus of control, or satisfaction with the doctor patient relationship. PMID- 11062967 TI - [Corneal infection in wearers of contact lenses: causes, effect on visual acuity and prevention]. AB - This is a 5-year retrospective survey of corneal infection in wearers of optical contact lenses (OCL). 23 of the 61 patients (38%; Hasharon Hospital) with positive cultures wore OCL. Visual acuity improved in 15 (65%), no change was noted in 4 (17.5%) and there was deterioration in 4 (17.5%), as compare with status on admission. Pseudomonas aeruginosa was the most common cause of infections among OCL wearers. The improvement in visual acuity expected due to wearing OCL was affected by infections. Those after Staphylococcus albus infections had the highest rate (100%) of improvement in visual acuity and after Ps. aeruginosa the lowest rate (57.2%) of improvement, as well as the highest rate of deterioration (42.8%) found following recovery. OCL wearers are at higher risk for damage to visual acuity following corneal infection, and highly virulent infections in OCL wearers are responsible for a high risk of damage to visual acuity. PMID- 11062968 TI - [Life-threatening echovirus 11 infection during first month of life]. AB - Infection with Echovirus 11 is mostly asymptomatic, but it may cause a wide variety of clinical diseases, from gastroenteritis to serious diseases such as meningitis and myocarditis. In small infants, especially during the first days of life, echovirus infection may appear as a sepsis-like illness, and cause disseminated intravascular coagulopathy and shock. We present 2 infants with severe Echovirus 11 infections. A 3.5-month-old died within 24 hours of shock and probably myocarditis. The other, 6-days old, presented with meningitis, hepatitis and disseminated intravascular coagulopathy. It recovered after treatment with intravenous immunoglobulin. Echovirus 11 may cause life-threatening infections in small infants. Pediatricians should be alert to the special characteristics of this disease. PMID- 11062969 TI - [Preservation of the larynx in advanced cancer--experiences at the Oncology Department, Rambam Medical Center, Haifa]. AB - The effectiveness of sequential chemo-radiotherapy in preserving the larynx in advanced laryngeal carcinoma was assessed. 4 unselected patients (19 men and 2 women, mean age 60 years) with advanced squamous cell carcinoma of the larynx (T3 4/N0-3) received induction chemotherapy consisting of 2-3 cycles of cisplatin (100 mg/m2) and 5-fluorouracil (1000 mg/m2/day) as a continuous infusion on days 1-5, followed by definitive radiotherapy: 50 Gy to the whole neck, 70 Gy to the larynx and clinically involved nodes, using a combination of 6 MV photons and 9 12 MeV electrons. 19 of the 21 patients responded to combined therapy but there was no response to induction therapy in 2 (10%) and 2 did not complete therapy due to severe toxicity. At a mean follow-up of 40 months, 7 had undergone total laryngectomy (33%), for an overall 5-year laryngeal preservation rate of 66%. Reasons for total laryngectomy in 2 patients were no response, and in 5 tumor recurrence. Mean survival was 39 months (range 11-46 months); at last follow-up, 17 of 21 were alive and disease-free, 11 of whom had a functional larynx (65% of survivors). 2 had died due to disease progression and due to a cardiovascular event. Sequential chemo-radiation allows laryngeal preservation in about 2/3 of surviving patients without compromising survival. PMID- 11062970 TI - [Combined approach in management of penetrating injury of vertebral artery]. AB - Penetrating injuries of the vertebral artery are not common. Although surgical control of this type of injury is well-documented, the combined approach (surgery and packing for temporary control of bleeding, followed by angiography and embolization for permanent control) is now recognized and practiced. We describe a 40-year-old man who was stabbed in the neck. He was rushed to the operating theater in hemorrhagic shock. A vertebral artery injury was identified and packed. Angiography and embolization permanently controlled the bleeding. The combined approach is safe and we recommend it in those in whom bleeding from the vertebral artery is initially well controlled with packing. PMID- 11062971 TI - [Dermatitis from contact with Agave americana]. AB - Various plants induce dermatitis in man. There have been only a few published cases of contact dermatitis caused by Agave americana (AA). We report intentional exposure to AA in a soldier seeking sick leave, and review our previously reported cases. Treatment with oral antihistamines and topical saline compresses resulted in subsidence of the systemic symptoms within 24 h and regression of cutaneous manifestations in 7-10 days. Physicians should be alert to the possibility of self-inflicted contact dermatitis induced by exposure to plants, especially to A. americana. Systemic signs may accompany the cutaneous lesions. PMID- 11062972 TI - [Thrombocytopenic purpura as sole manifestation of brucellosis in children]. AB - Thrombocytopenic purpura associated with brucellosis has rarely been described in children. The thrombocytopenic purpura is usually part of the array of manifestations of brucellosis, such as fever, malaise, arthralgia, arthritis, hepatosplenomegaly and lymphadenopathy. We describe a 4-year-old girl in whom severe thrombocytopenic purpura was the only manifestation of brucellosis and resolved after appropriate antibiotic therapy. We conclude that brucellosis should be included in the differential diagnosis of thrombocytopenic purpura in areas endemic for brucellosis, and when there is a history of exposure to infected food products. PMID- 11062973 TI - [Passive smoking and its influence on the eye]. PMID- 11062974 TI - [Anti-angiogenesis drugs as a potential approach in cancer therapy]. PMID- 11062975 TI - [Apoptosis during normal and abnormal embryogenesis]. PMID- 11062976 TI - [Thrombocytopenia in pregnancy]. PMID- 11062977 TI - [Eating disorders in athletes: myths and warning signs]. PMID- 11062978 TI - [Orthopedic aspects of Rett's syndrome]. PMID- 11062979 TI - [Suicide from the Jewish viewpoint]. PMID- 11062980 TI - [Jewish nurses and doctors as victims of personal terror during the British mandate]. PMID- 11062981 TI - [Virginity and defloration]. PMID- 11062982 TI - [Henry Lewis Barnett]. PMID- 11062983 TI - [Physicians and the Holocaust]. PMID- 11062984 TI - Identification and characterization of a cDNA for mouse ameloblastin. AB - Ameloblastin was first identified as one of the most abundant novel transcripts from a random screening of a rat incisor cDNA library. In situ hybridization experiments have shown ameloblastin expression to be specific to ameloblasts, with highest levels in secretory and maturation stage ameloblasts and cells of the epithelial root sheath. Ameloblastin has been identified as a candidate gene for the local hypoplastic form of autosomal dominant amelogenesis imperfecta, by virtue of it's location within the critical disease locus. The purpose of this study was to isolate a full length mouse ameloblastin cDNA and determine its temporal expression pattern during odontogenesis. A newborn mouse molar cDNA library was screened using a rat ameloblastin cDNA probe. Positive clones were confirmed by PCR analysis with ameloblastin-specific primers, and their size determined with vector-specific primers. Phage clones were rescued to phagemid using Exassist helper phage and the nucleotide sequence determined. We report here the identification of two clones, exhibiting alternative splicing of the putative open reading frame, and use of multiple polyadenylation signals. Nucleotide sequence analysis indicated a high degree of similarity to rat ameloblastin, rat amelin 1 and 2 and porcine sheathlin. Reverse transcriptase-PCR analysis using mouse first and second mandibular molar mRNA indicated initial expression at E-14. This is one day after the initial expression of tuftelin (E 13) and one day prior to that of amelogenin (E-15). PMID- 11062985 TI - Molecular cloning and characterization of the bovine and human tuftelin genes. AB - The bovine tuftelin gene was cloned and its structure determined by DNA sequence analysis and comparison to bovine tuftelin cDNA. The analyses demonstrated that the cDNA contains a 1014 bp open reading frame encoding a protein of 338 residues with a calculated molecular weight of 38,630 kDa and an isoelectric point of 5.85. Although similar, these results differ from those previously published [Deutsch et al. (1991) J. Biol. Chem. 266, 16021-16028] which contained a different conceptual amino acid sequence for the carboxy terminal region and identification of a different termination codon because of the absence of a single guanine residue in the published sequence. The protein does not appear to share homology or domain motifs with any other known protein. The bovine gene consists of 13 exons ranging in size from 66 to 1531 bp, the latter containing the encoded carboxy terminal and 3' untranslated regions. These exons are embedded in greater than 28 kbp of genomic DNA and codons are generally not divided at exon/intron borders. Sequence analysis of the cDNA and products produced by reverse transcriptase/polymerase chain reaction demonstrated that exons 2, 5 and 6 are alternatively spliced. The 3' portion of the human gene was also isolated and characterized by DNA sequencing, which demonstrated agreement between the bovine and human sequences in the segment in question. The difference between the presently reported sequence and that of the previously published one suggests the possibility of an unusual type of polymorphism which would result in markedly different amino acid sequences at the carboxy terminal region of the protein. The human tuftelin gene was localized to chromosome 1q21 by in situ hybridization. PMID- 11062986 TI - Developmental regulation of dentin sialophosphoprotein during ameloblast differentiation: a potential enamel matrix nucleator. AB - The two major dentin matrix proteins, dentin sialoprotein and dentin phosphoprotein have been shown to be expressed as a single large transcript termed dentin sialophosphoprotein (DSPP). These non-collagenous matrix proteins, identified biochemically by their unique physical-chemical properties, are specific cleavage products of a large parent acidic phosphorylated protein (pI 4.0). Previous studies have shown expression of dentin sialoprotein at the protein level by ameloblasts. The purpose of this study was to determine the temporal-spatial pattern of DSPP expression during amelogenesis. In situ hybridization and immunohistochemistry were performed on sections of developing mouse molars. These data were correlated with RT-PCR analysis of in vitro enamel organ epithelium monolayer cell cultures enriched for ameloblasts. Our data indicates initial expression of the DSPP transcripts and protein during early ameloblast differentiation prior to the secretory phase when the majority of the enamel matrix is formed. Ameloblasts appear to tightly down-regulate DSPP transcription as enamel matrix formation is up-regulated. These data demonstrate DSPP expression during amelogenesis is under highly controlled developmental regulation. Therefore, DSPP may have a primary role in the initial mineralization events of both enamel and dentin, acting as a potential nucleator of hydroxyapatite crystal formation. PMID- 11062987 TI - Sites of asparagine-linked oligosaccharides in porcine 32 kDa enamelin. AB - The 32 kDa enamelin protein isolated from developing porcine enamel was previously shown to contain eight different asparagine-linked oligosaccharides. However, only three consensus attachment sites were evident in this protein. In this study, glycopeptides containing all three potential glycosylation sites (72 Asn, 79-Asn and 91-Asn) were purified from 32 kDa enamelin. The oligosaccharides were isolated from each glycopeptide following digestion with N-oligosaccharide glycopeptidase, labeled with 2-aminopyridine at the reducing ends, and then characterized by reverse phase HPLC. All three potential sites were found to be glycosylated heterogeneously (i.e., five biantennary complexes at 72-Asn, two biantennary complexes at 79-Asn, three triantennary complexes at 91-Asn), accounting for all eight oligosaccharides characterized previously. These results indicate that 32 kDa enamelin has a complex pattern of asparagine-linked glycosylation localized within a small region (20 residues) of the protein. The functional significance of this glycosylation remains to be established. PMID- 11062988 TI - Murine enamelin: cDNA and derived protein sequences. AB - Enamelin is the largest enamel protein. Recently we reported the characterization of a cDNA clone encoding porcine enamelin. The secreted protein has 1104 amino acids--over 6 times the length of amelogenin (173 amino acids) and almost 3 times the lengths of sheathlin (395 amino acids) and tuftelin (389 amino acids). Immunohistochemistry has shown that uncleaved porcine enamelin concentrates at the growing tips of the enamel crystallites while its cleavage products localize to rod and interrod enamel. Here we report the isolation and characterization of cDNA encoding murine amelogenin and demonstrate the tooth specificity of porcine enamelin. The murine clone is 4154 nucleotides in length and encodes a protein of 1274 amino acids. In the absence of post-translational modifications murine enamelin has an isotope averaged molecular mass of 137 kDa and an isoelectric point of 9.4. Multiple tissue Northern blot analyses detect porcine enamelin mRNA in developing teeth but not in liver, heart, brain, spleen, skeletal muscle and lung. Mouse and porcine enamelin share 61% amino acid identity and 75% DNA sequence identity. Mouse enamelin has 14 tandemly arranged copies of an 11 amino acid segment that is found only once in porcine enamelin. PMID- 11062989 TI - Role of matrix proteases in processing enamel proteins. AB - This article reviews the current status of research on proteases of the enamel layer that are capable of processing and degrading proteins of the enamel matrix. Following a brief survey of the historical development of this discipline, a summary is presented of the current status. Two proteases have recently been cloned: EMSP-1 (enamel matrix serine protease-1), a serine protease, and enamelysin, a metalloprotease. These two are placed into their appropriate families: the chymotrypsin family S1 of clan SA of the serine protease class and the matrixin family or matrix metalloproteinase family, M10 of clan MB (the metzincins) of the metalloprotease class. The major features of these two families are outlined. The article concludes with some suggested areas for future research--identifying further proteases and characterizing those now known. PMID- 11062990 TI - Degradative changes in whole enamel homogenates incubated in vitro in the presence of low calcium ion concentrations. AB - The purpose of this study was to investigate overall degradative changes occurring to enamel matrix proteins in small, freeze-dried pieces of rat incisor enamel homogenized and incubated directly for 0-48 hours in a synthetic enamel fluid solution (165 mM total ionic strength with 0.153 mM calcium chloride) versus other samples homogenized and incubated for the same time intervals in distilled water. The results indicated that many alterations in the apparent molecular weights of enamel matrix proteins took place under both conditions although the rates for many degradative changes over a 48 hour period were often slower in distilled water than in synthetic enamel fluid. Freeze-dried enamel samples homogenized and incubated in 165 mM Tris-HCl buffer at pH 8.0 showed changes comparable to those seen with distilled water. This suggested that differences observed between samples incubated in enamel fluid versus distilled water were unrelated to pH or ionic strength of the solutions and may be the result of a requirement by some enamel proteinases for small amounts of free calcium ions in incubation media. Of interest were findings that some enamel matrix proteins, especially those in strips taken from the first half of the secretory stage of amelogenesis, were degraded much faster in distilled water than in synthetic enamel fluid. The reasons for this effect are unclear although, in this case, calcium ions could be inhibitory to hydrolysis of certain matrix proteins by the enamel proteinases. PMID- 11062991 TI - Enzyme compartmentalization during biphasic enamel matrix processing. AB - Processing of enamel matrix proteins is essentially biphasic. Secretory stage metalloprotease activity generates a discrete, presumably functional, spectrum of molecules which may also undergo dephosphorylation. Maturation stage serine proteases almost completely destroy the matrix. The present aim was to examine the tissue compartmentalization of these enzyme activities in relation to their possible function. A sequential extraction using synthetic enamel fluid, phosphate buffer and SDS was used to identify enzymes free in the enamel fluid, crystal bound or aggregated with the bulk matrix respectively. Results indicated that the metallo-proteases and alkaline phosphatase were free in the secretory stage enamel fluid while the serine proteases appeared to be largely bound to the maturation stage crystals. The mobility of the metallo-proteases and alkaline phosphatase would ensure efficient initial processing of secretory matrix, while the largely mineral bound serine proteases would ensure retention of protease activity despite massive destruction and protein removal. PMID- 11062992 TI - Enamelysin mRNA displays a developmentally defined pattern of expression and encodes a protein which degrades amelogenin. AB - Previously, a cDNA encoding a novel matrix metalloproteinase (enamelysin) was isolated from a porcine enamel organ-specific cDNA library. The cloned mRNA is tooth-specific and contains an open reading frame encoding a protein composed of 483 amino acids (Gene, 183:(1-2), p123-128, 1996). Here, we show that: 1) The expression of enamelysin mRNA is not limited to the enamel organ as previously reported. The enamelysin message is also expressed at very low levels in the pulp organ. 2) Northern analysis reveals that the enamelysin mRNA displays a developmentally defined pattern of expression in the enamel organ. The message is expressed at relatively high levels during the presecretory and early transition stages of development. However, during late maturation, the quantity of enamelysin mRNA is greatly reduced. Conversely, the low message levels in the pulp organ remain relatively constant throughout these developmental stages. 3) The enamelysin cDNA was ligated into a prokaryotic expression vector and recombinant enamelysin containing a His tag was purified from E. coli. Zymographic analysis utilizing recombinant murine amelogenin as the substrate, reveals that the purified enamelysin degrades amelogenin. Since enamelysin is developmentally regulated and is capable of degrading amelogenin, it is likely to play a significant role during enamel biomineralization. PMID- 11062993 TI - Enamel matrix serine proteinase 1: stage-specific expression and molecular modeling. AB - Enamel proteins are cleaved by proteinases soon after their secretion by ameloblasts. Intact proteins concentrate in the outer enamel at or near the growing tips of the enamel crystallites while cleavage products accumulate in the deeper enamel. In the transition and early maturation stages there is a dramatic increase in proteolytic activity. This activity, coupled with the diminished secretory and increased reabsorptive functions of ameloblasts, leads to a precipitous fall in the amount of enamel protein in the matrix. Recently we have cloned and characterized an mRNA encoding a tooth-specific serine proteinase designated enamel matrix serine proteinase 1 (EMSP1) [Simmer et al., JDR (1998) 77: 377]. EMSP1 can be detected in the inner enamel during the secretory stage and its activity increases sharply during the transition stage. Stage-specific Northern blot analysis demonstrates this increase is accompanied by a parallel increase in the amount EMSP1 mRNA. A 3-dimensional computer model of EMSP1, based upon the crystal structure of bovine trypsin, has been generated and analyzed. All six disulfide bridges as well as the active site are conserved. Changes in the peptide binding region and the specificity pocket suggest that interaction of the proteinase with protein substrates is altered, potentially causing a shift in substrate specificity. The calcium binding region of trypsin is thoroughly modified suggesting that the calcium independence of EMSP1 activity is due to an inability to bind calcium. The three potential N-linked glycosylation sites, N104, N139 and N184, are in surface accessible positions away from the active site. PMID- 11062994 TI - Degradation of enamel matrix proteins in porcine secretory enamel. AB - To elucidate the progressive disappearance of 25 kDa amelogenin occurring in a narrow space near the surface of enamel, the alkaline soluble fraction which contained 80% of the total proteins was extracted from a newly formed porcine enamel. When this fraction was incubated with the addition of Ca ions in an in vitro system, the degradation of the coexisting amelogenin and enamelin occurred without activation during the incubation period. Although the fraction contained mainly two kinds of metalloproteinases, 56 kDa and 61 kDa gelatinolytic, and 41 kDa and 46 kDa caseinolytic activities, it was demonstrated on amelogenin enzymography that the caseinolytic one was concerned with the conversion of the 25 kDa amelogenin into the 20 kDa amelogenin. The protein distribution of the newly formed enamel indicated that the metalloproteinases degraded the coexisting enamelin and amelogenin imperfectly. Nevertheless, during the next developing stage they demonstrated their full activities. It is suspected that these activities are regulated by Ca ions, which may be increased by a cascade system. PMID- 11062995 TI - Effect of apatite crystals on the activity of amelogenin degrading enzymes in vitro. AB - The objective of the present study was to determine the effect of apatite crystals on the activity of amelogenin degrading enzymes in vitro. Current experimental data, together with previous reports support the view that among the different proteinases present in the enamel extracellular matrix, serine proteinase(s) are responsible for the massive degradation of amelogenins during the maturation stage. For our in-vitro experiments we used the recombinant amelogenin M179 as substrate and a "65%-satd. (NH4)2SO4" fraction of enamel proteins as well as chymotrypsin as sources for serine-proteinase activity. We report preliminary experiments of amelogenin proteolysis in the presence of apatite crystals resulting in a different proteolysis pattern when compared to amelogenin proteolysis without apatite crystals. Quantitative analysis of the HPLC peaks corresponding to the proteolysis products indicates that the presence of apatite crystals in the proteolysis solution inhibits the ability of the serine-proteinases to degrade amelogenin. The present observations support the hypothesis that amelogenin degradation correlates with apatite crystal growth during enamel maturation. PMID- 11062996 TI - The aetiology of developmental defects of enamel: a prevalence and family study in East London, U.K. AB - The aim of this study was to investigate genetic and environmental factors associated with hypoplastic defects of enamel in a detailed family study combined with an overall prevalence study. 68 percent of 1,518 children with a "complete" permanent dentition had enamel defects, 14.6 percent having hypoplasia. The Family Study consisted of 101 Index Cases from the Prevalence Study having 2 or more teeth with hypoplasia and their first degree relatives: they were compared with 101 matched controls and their relatives. The clinical examinations were supplemented with structured interviews to obtain social, medical and dental histories. Three Index Cases had amelogenesis imperfecta and 18 had "chronological hypoplasia". Bilateral hypoplasia of lower central incisors had a multifactorial aetiology (heritability 70 percent +/- 38 percent), while hypoplasia of pre-molars was associated with familial occurrence of defects. There was evidence suggesting a predisposition in some families to developmental defects of enamel. PMID- 11062997 TI - SEM study of the development of rat incisor enamel hypoplasia under hypocalcemia induced by thyro-parathyroidectomy. AB - Several clinical studies have reported the presence of enamel hypoplasia in congenital hypoparathyroidism or hypocalcemia. In previous studies we showed that thyroparathyroidectomy (TPTX) induced perturbations of the ameloblast morphology and secretion, of the rod pattern and of the enamel surface at late secretory stage and beginning of maturation, and limited hypoplasia in the erupted enamel of rat incisor. The aim of the present study was to evaluate by SEM, the extent and evolution of the enamel alterations of thyro-parathyroidectomized rats during the maturation stage. Wistar rats were thyro-parathyroidectomized and sacrificed 57 days later. The incisors were dissected out and processed for SEM. The surface of the incisor was observed from the end of secretion/beginning of maturation to its incisal erupted end. Transverse sections were prepared to study the structural defects and the prism pattern at different stages. The results showed that the surface of the TPTX incisors presented large hypoplastic defects at the end of secretion/beginning of maturation and only small defects in the erupted part. Transverse sections showed that, at the transition from secretion to maturation, the enamel defects extended to the mid-thickness of the tissue. At the incisal end the defects were limited to the outer enamel. As it is difficult to understand how the large apical defects could recover to appear as small hypoplasia at the incisal end, these results raise new questions concerning: (1) the effect of a long term calcium deficiency upon the cellular activity of the ameloblasts, and (2) the capacity of the enamel organ to compensate structural abnormalities. PMID- 11062998 TI - Enamel structure and composition in the tricho-dento-osseous syndrome. AB - Tricho-dento-osseous syndrome (TDO) is an autosomal dominant disorder characterized by curly hair, hypoplastic enamel, taurodontism, and dense bone. The purpose of this investigation was to characterize the enamel defects in a TDO population in North Carolina. Twelve TDO teeth and 12 normal teeth were examined. The enamel thickness was decreased in all TDO teeth ranging from having no enamel to about 60% the thickness of normal teeth. Half of the TDO teeth had primarily prismless enamel while the remainder had at least occasional areas of prismatic enamel. TDO enamel crystallites appeared similar to normal crystallites with TEM. The mineral per volume of TDO enamel (n = 9) (68.5%) was significantly less, on average, compared with normal enamel (n = 8) (84.5). The genetic mutation responsible for the TDO phenotype results in alteration of a developmental pathway(s) common to hair, teeth and bone. This further illustrates that these embryologically diverse tissues share common developmental controls at the molecular level. PMID- 11062999 TI - Tuftelin mRNA is expressed in a human ameloblastoma tumor. AB - RT-PCR, Southern blotting and DNA sequencing have established for the first time that tuftelin mRNA is expressed in human ameloblastoma tumor. The expression of amelogenin mRNA in ameloblastoma was also established, confirming earlier reports by Snead et al. These results corroborate, on a molecular level, the enamel organ epithelial origin of ameloblastoma. In view of the present results, it is interesting that previous studies have indicated that although ameloblastoma, a non-mineralized odontogenic tumor, transcribes amelogenin mRNA, amelogenin (and enamelin) proteins are not expressed in this tissue. However, in mineralizing odontogenic tumors, both these classes of proteins are expressed. PMID- 11063000 TI - Evolutionary and functional significance of hominoid tooth enamel. AB - The purpose of this investigation is to evaluate enamel thickness in extant and extinct hominoids. The material used in this study spans the evolutionary history of this group, from 20 million years ago to the present. The objectives of this investigation are to test three hypotheses: (1) the Loading Hypothesis: loading areas of the crown have thicker enamel than non-loading areas; (2) the Phyletic Hypothesis: differences in enamel thickness provide a basis for determining evolutionary relationships; and (3) the Functional Hypothesis: differences among hominoids result from adaptations to differing dietary and ecological habitats, that is from folivory to frugivory to hard object feeding and from tropical to forest to savanna habitats. Thin sections were prepared and polished to approximately 100 microm in thickness. Each section was then enlarged and digitally captured to the computer. Image processing and analysis software, SigmaImage (was used to measure the sections. Subsequent statistical analysis was conducted with SigmaStat and SPSS statistical software programs. The data provides statistical support for all hypotheses. In particular, the data support the proposal that "thick" enamel is the ancestral condition for the great apes and human clade. Therefore, Pongo would have retained its enamel thickness from the common ancestor of the great apes and Gorilla and Pan would have secondarily reduced enamel thickness to "thin." The common ancestor of the hominids, the australopithecines, would have "thick" enamel. The "hyper-thick" enamel of the australopithecines would be a derived character for this clade due to increased crushing and grinding and adaptation to savanna habitat. Homo would have secondarily reduced enamel thickness to "thick." Evolutionary biology of enamel differs markedly in hominids from that found in other hominoids and primates. Increased enamel thickness involved both increases in absolute thickness of enamel and crown size in response to increase masticatory loading. PMID- 11063001 TI - Mechanisms of mineralization in the enameloid of elasmobranchs and teleosts. AB - Ultrastructural and cytochemical studies on the mineralization of enameloid were performed using Heterodontus japonicus, an elasmobranch, and Tilapia buttikoferi, a teleost as materials. The mineralization of the enameloid in the Heterodontus was divided into the following two steps: (1) initial crystallization in the tubular vesicles that originated from the odontoblasts, and (2) crystal growth that was accompanied by the degeneration and removal of the organic matrix around the crystals. In the Tilapia, the mineralization of the cap enameloid followed three steps: (1) initial crystallization at the matrix vesicles, (2) aggregation of fine slender crystals along collagen fibrils, and (3) crystal growth with the degeneration and removal of the organic matrix. The pattern of early mineralization and the composition of organic matrix in enameloid were considerably different between the two species examined, while in both species the odontoblasts were mainly involved in the formation of the organic matrix of enameloid and in the initial mineralization. In the next step, remarkable crystal growth associated with the degeneration and removal of the organic matrix occurred in both the elasmobranch and the teleost species. The absorptive functions of the dental epithelial cells in the later stages of enameloid formation is probably similar in the two types of enameloid, and is essential for the production of well-mineralized enameloid. PMID- 11063002 TI - The function and structure of the marsupial enamel. AB - The aims of this study are to clarify the structure of tubular enamel and the function of enamel tubules on the marsupial of opossum (Monodelphis domestica). Almost all enamel prisms, surrounded by interprismatic enamel, ran obliquely from the dentinoenamel junction (DEJ), and bent near the enamel surface. The enamel tubules are distributed in both enamel prisms and the interprismatic enamel near the DEJ. From the middle to the surface of the enamel, one enamel tubule ran within a single enamel prism. Most of enamel tubules continued from the DEJ to near the enamel surface. It is suggested that each enamel tubule developed in relation to one ameloblast. The fibers of odontoblastic process penetrated the DEJ from the dentinal tubules into the enamel tubules, and some branched across the enamel prisms. The odontoblastic process may be actively cross into the ameloblastic layer and may be involved in the formation of enamel tubules. After in vivo injection of tetracycline, tetracycline labeling showed that the odontoblastic tubules continued to enamel tubules. And strontium was detected in enamel tubules from the DEJ to the enamel surface, as was the dentinal tubules. In conclusion, there was active transport by the odontoblast and it's process through the enamel tubules. PMID- 11063003 TI - Closing panel discussion. What have we learned? New directions in enamel research. PMID- 11063004 TI - COMP (cartilage oligomeric matrix protein) is synthesized in ligament, tendon, meniscus, and articular cartilage. AB - The presence of cartilage oligomeric matrix protein (COMP) in extracts of ligament, tendon, meniscus, and canine articular cartilage was demonstrated by Western blot analysis using anti-dog COMP antibody. When the tissues were cultured in the presence of [35-S]methionine/cysteine, metabolically labeled COMP was purified from the culture media and from tissue extracts by DEAE-cellulose gel chromatography. SDS-Polyacrylamide gel electrophoresis (SDS-PAGE) followed by autoradiography and immunoblotting under reducing and non-reducing conditions revealed that COMP is synthesized by the cells of these connective tissues. Increased levels of COMP in samples of both synovial fluid and serum of patients with various joint diseases may not only be derived from cartilage but also from ligaments and tendons. COMP is not a highly tissue-specific cartilage molecule. PMID- 11063005 TI - Sustained loading increases the compressive strength of articular cartilage. AB - INTRODUCTION: When synovial joints are subjected to sustained or repetitive loading, fluid is driven from the articular cartilage so that it is less able to equalise compressive stress between opposing joint surfaces. We test the hypothesis that sustained loading reduces the compressive strength of cartilage on-bone. METHODS: Forty specimens of articular cartilage-on-bone, approximately 15 mm square, were removed from the patella groove of mature bovine knees. Specimens were loaded on a materials testing machine using a 5 mm-diameter plane ended indentor. Controlled loading/unloading cycles of 1s duration, and of increasing severity, were applied until failure was evident on the force deformation graphs. Half of the specimens were 'creep loaded' for 30 at 2 MPa before their strength was assessed. After testing, damage was investigate using ink staining of the cartilage surface, and histology. RESULTS: Sustained loading reduced cartilage thickness by 45% and creep-loaded specimens were 21% stronger (P = 0.01). Most specimens appeared to fail by fissuring of the cartilage surface zone. CONCLUSION: Sustained loading strengthens cartilage by expelling water from it, reducing the tendency of the surface zone to rupture in the manner of an over inflated car tyre. PMID- 11063006 TI - Expression of type I and type V collagen mRNAs in the elasmoid scales of a teleost fish as revealed by in situ hybridization. AB - The ability of scale-forming cells to produce both type I and type V collagens was investigated by in situ hybridization at the light and electron microscope levels. Biochemical analyses reported that type I collagen, the predominant component, was associated with the minor type V collagen in the collagenous matrix of the teleost scales where, thin and thick collagen fibrils formed distinct layers. Thin collagen fibrils of the external layer were produced by the episquamal scleroblasts scattered on the outer scale surface, while thick collagen fibrils forming the compact basal plate were produced by the hyposquamal scleroblasts lining the inner surface of the scale. We demonstrated that episquamal and hyposquamal scleroblasts contained mRNAs for alpha1(I) and alpha1(V) collagens. Quantification by image analysis of the relative amount of alpha1(I) and alpha1(V) mRNAs in episquamal and hyposquamal scleroblasts suggests that the gene expression of type V collagen was proportionally higher in episquamal scleroblasts. These results support our hypothesis that the diameter of the thin fibrils of the external layer is regulated by the significant amount of type V collagen that interacts with type I collagen. PMID- 11063007 TI - Human root dentin: structural anisotropy and Vickers microhardness isotropy. AB - The demanding mechanical functions and the variable structure of dentin make it an invaluable material for studying the structure-mechanical function relations of a mineralized collagen-containing tissue. The mineralized collagen fibril axes in human root dentin are mainly located on the incremental plane. Within this plane there is a preferred orientation in the general root-crown direction. The apatite crystals are aligned in three dimensions within an individual collagen fibril, but this orientation does not necessarily extend to the neighboring fibrils. Crystals are also present as aggregates without any preferred orientation. The structure is therefore clearly anisotropic with respect to the collagen fibril orientation, but less so with respect to overall crystal orientation. Vickers microhardness measurements of the root dentin are essentially the same on the three orthogonal planes with respect to the incremental plane. Knoop microhardness measurements are also the same on all three orthogonal planes when the major diagonal is aligned perpendicular to the collagen fibril axis preferred orientation direction. In-plane variations of up to 20% are observed in the orthogonal direction. The material is thus isotropic in the three main directions with respect to Vickers microhardness, but anisotropic in structure. This paradoxical situation is attributed mainly to the variable modes of crystal organization. PMID- 11063008 TI - Type V collagen in experimental granulation tissue. AB - To evaluate the spatial and temporal expression of type V collagen in a wound healing model, subcutaneously implanted viscose cellulose sponges in rats were used to induce granulation tissue formation. Analyses on granulation tissue were carried out on days 3, 5, 8, 14, 21, 30, 59 and 84. Acid soluble collagens were extracted and the relative amount of type V collagen was quantified by SDS-PAGE. Specific antibodies to type I, III and V collagens were used in immunohistochemistry and specific RNA probes to proalpha1(I), proalpha1(III) and proalpha1(V) collagen in in situ hybridization. Type V collagen content increased relative to type I and III collagens up to day 8 and remained at the same level for up to the three months. Type V collagen was expressed strongly in blood vessel walls as seen in immunohistochemistry. In situ hybridization showed that all of the three types of collagen were expressed mostly in fibroblast-like cells and also in rounded cells, especially type V collagen. In conclusion, type V collagen was seen in the wound healing model in increasing amounts from day 3 onwards, its localization being highly associated with blood vessels in granulation tissue and it was synthesized by fibroblast-like and rounded cells. PMID- 11063009 TI - Thermal stability of bone collagen as an indicator of bone turnover in gonadectomized and multiparous rats. AB - Previous findings indicate that the thermal stability of bone collagen is related to age. In this study, collagen from rat bone with reported different turnover rates was investigated. Cortical and trabecular bone from femur were obtained from intact, ovariectomized, orchidectomized and multiparous breeder rats. Thermal stabilities of fibrillar collagen in decalcified bone matrix and molecular collagen obtained by pepsin treatment were measured as shrinkage (Ts) and 'melting' temperature (Tm), respectively. Both Ts and Tm of cortical collagen from intact female rats decreased in parallel with age as previously found in male rats indicating that Ts and Tm measurements are interchangeable techniques in characterizing the thermal stability of bone collagen. Tm of trabecular collagen from intact rats decreased with age, however, with a decay only one third of that for cortical collagen. The different rates possibly reflect different ages of collagen due to remodeling activity present in trabecular and minimal in cortical bone. Compared with control rats the Tm of trabecular collagen from gonadectomized and multiparous rats with a reported increased trabecular turnover rate was elevated, whereas only minor variations in Tm of cortical collagen were found. In conclusion, the thermal stability of bone collagen decreases with the age of the collagen. Increased bone turnover implies elevated thermal stability of bone collagen. Thus, thermal stability of bone collagen appears to be an indicator of bone turnover. PMID- 11063010 TI - Integrin expression is upregulated during early healing in a canine intrasynovial flexor tendon repair and controlled passive motion model. AB - To explore crucial early molecular events involved in contact healing of the intrasynovial flexor tendon, integrin expression was evaluated at the transcriptional and post-transcriptional levels during the first two weeks following injury, repair and controlled passive motion in a canine model. Specifically, immunohistochemical and reverse transcription polymerase chain reaction (RT-PCR) techniques were employed to evaluate expression of the fibronectin, vitronectin and endothelial cell binding integrin receptor subunits alpha5, alphav and alpha6, along with the common beta1 subunit. The two techniques revealed increasing expression of the four subunits over the two week post-repair period. Immunohistochemistry revealed that beta1 and alpha5 expression was concentrated in the epitenon layer near the repair site and interiorly within the wound area, while alpha6 was associated with capillary forming endothelial cells near the wound. RT-PCR and quantitation by NIH image analysis demonstrated peak messenger RNA expression of beta1 and alpha5 at ten days post-repair and peak expression of alpha6 and alphav at 15 days. The results in this study correlate well with previous results demonstrating increased fibronectin deposition and angiogenesis during the same time period in a similar injury/repair model. PMID- 11063011 TI - Primary culture and characterization of enamel organ epithelial cells. AB - The cells of the enamel organ are programmed by signals such as growth factors and extracellular matrix components to differentiate and form dental enamel. To study how the enamel organ epithelial cells control enamel development, we have begun to characterize a primary porcine enamel organ epithelial cell culture system. The unerupted molars of 3 month old pigs were isolated, the cells were digested into a single cell suspension and grown in media either with or without serum. Expression of amelogenin and ameloblastin mRNA was monitored by RT PCR, and protein secretion was identified by immunohistochemistry. Cells grown in MEM formed a mixed cell population of epithelial- and fibroblast-like cells which grew past confluence, formed nodules, mineralized, and expressed low levels of amelogenin and ameloblastin protein. In LHC-9 media, which is selective for epithelial cells, the cells did not grow past confluence but secreted amelogenin and ameloblastin proteins more strongly. Cell viability was maintained in both serum-free and serum-containing media. However, in the serum-free media, cell proliferation proceeded slowly. Although cells grown in MEM mineralized, the mixed cell population may make studies of specific ameloblast-like cells more difficult. However, cells grown in a culture media selective for epithelial cells will require modifications such as cell immortalization to allow long term studies of cell regulation and interaction. In summary, we have established an enamel organ epithelial cell culture system which will enable us to study the role of ameloblasts in enamel matrix formation, ameloblast regulation, as well as cell-matrix interactions. Selection of specific culture conditions will depend on the questions being addressed in individual studies. PMID- 11063012 TI - Isolation of amelogenin-positive ameloblasts from rat mandibular incisor enamel organs by flow cytometry and fluorescence activated cell sorting. AB - The purpose of this study was to use amelogenin as a marker to examine the feasibility of isolating ameloblasts from enamel organ cell populations by fluorescence activated cell sorting. After treating dissected rat enamel organs with proteolytic enzymes to loosen cell attachments and labial connective tissues, dissociated cell suspensions were fixed, then immunostained with rabbit anti-rM179 recombinant amelogenin antibody and FITC-conjugated goat anti-rabbit Ig G antibody. Flow cytometry indicated that about 70% of the total cell sample and virtually all the larger cells therein were amelogenin-positive. Fluorescence activated cell sorting yielded a sample of amelogenin-positive cells at 97% purity. Immunofluorescence microscopy indicated that these isolated amelogenin positive cells varied widely in size and morphology. This was attributed to loss of intercellular support for ameloblasts once they were dissociated from each other, and to some fragmentation caused when the cells were initially physically removed from the teeth. The results demonstrate that viable ameloblast cell fractions, especially representing cells at the secretory stage, can be purified from enzymic digests of rat enamel organ by sorting on the basis of cell size alone. From these fractions, subpopulations of ameloblasts may be identified when differentiation specific cell surface markers become available. PMID- 11063013 TI - Enamel cell biology. Towards a comprehensive biochemical understanding. AB - Enamel cells ultimately determine the properties of dental enamel. Surprisingly little is known about enamel cell functions at the biochemical and molecular levels. Understanding of both normal and abnormal enamel formation should benefit from elucidation of this area. This paper reviews our recent efforts to establish microscale biochemical analyses of rat enamel cells, and the ensuing initial findings about their protein phenotype (i.e., proteome) and calcium-handling mechanisms. A perspective of the current status of enamel cell research, and where it might head, is also given. PMID- 11063014 TI - Origin of enamel prisms and Hunter-Schreger bands in reptilian enamel. AB - Enamel prisms in mammalian enamel structure including human are considered to have originated in the reptilian enamel. The author has reported the original pattern of enamel prisms and of Hunter-Schreger bands in fossil reptiles of Mosasaurus sp. and Phytosaurus sp. The origin of the patterns of crystallite groups appear to arise in the fossil reptiles. Mosasaurus sp. had an island pattern and Phytosaurus sp. had a slender pattern. These two types are of various sizes, and are formed by variable numbers of ameloblasts. The number of ameloblasts involved in the formation of the crystallite group may change during amelogenesis. The present study observed the initial crystallite groups as enamel islands, in the surface of the early secretory stage of amelogenesis of living Alligator mississippiensis. These crystallite groups become bigger and more uniform in size during amelogenesis, being formed by several ameloblasts in the maturation stage. The border of crystallite groups observed appear as the lamella in human enamel. These crystallite groups may develop into the enamel prisms and Hunter-Schreger bands. PMID- 11063015 TI - Relationship between the electrical resistivity of enamel and the relative humidity. AB - The present study was carried out to investigate the relationship between the relative humidity and the electrical resistivity of enamel in vitro. The electrical resistivity of enamel samples was measured in a thermo-hygrostat where the relative humidity varied between 60% and 90%, with the measurements carried out first in a descending direction and then in an ascending direction. When the electrical resistivity was measured from a low to high relative humidity in an ascending direction, a higher value of electrical resistivity was always obtained in comparison to that when measured in the reverse descending direction. The difference between the mean electrical resistivity in an ascending and a descending direction increased in proportion to the relative humidity. These findings suggest that the diffusion of electrolytes and water in enamel is sensitive to changes in the relative humidity. PMID- 11063016 TI - Determination of mineral concentration in dental enamel from X-ray attenuation measurements. AB - The mineral content of dental enamel is commonly measured by X-ray attenuation experiments. Most studies have used contact microradiography in which intensities are measured with photographic film which is convenient and gives high spatial resolution. However photon counting intensity measurements are to be preferred in many experiments (longitudinal and scanning microradiography, and microtomography), as illustrated here, because they have a larger dynamic range and greater sensitivity to small intensity changes. Additionally, the detector and specimen are well separated which allows the pseudo-continuous study of de- and remineralization. The mineral content is often quoted as 95 wt% or 87 vol% hydroxyapatite for permanent human enamel. This determination from attenuation experiments requires accurate values of elemental mass attenuation coefficients and a number of assumptions. The effects of possible choices of these are considered and it is shown that the most important is the density of enamel mineral used in conversion of wt% to vol%. If the density is taken as 2.99 g cm( 3), as recently suggested (J.C. Elliott, Dental Enamel, Ciba Foundation Symposium 205, Wiley, Chichester, pp. 54-72, 1997), instead of 3.15 g cm(-3) as for hydroxyapatite, the calculated vol% is approximately 93 instead of approximately 87. PMID- 11063017 TI - Differences in co-variation of inorganic elements in the bulk and surface of human deciduous enamel: an induction analysis study. AB - This paper demonstrates a method for determination of co-variation between some inorganic elements in the bulk and surface areas of human tooth enamel. The technique is based on a computerised induction analysis of data obtained by secondary ion mass spectrometry (SIMS). When comparing the present data with an earlier study from our laboratory, it became evident that with only a moderate increase in the amount of data for the induction analysis, the results increased very considerably in reliability and precision. The patterns of co-variation between different elements differed between the bulk- and surface enamel. Only Mg expressed a similar pattern. In the bulk enamel the elements Na and P expressed a high degree of co-variation. Similarly, Mg and C expressed a high degree of co variation. Mg was an element often found to co-vary with bulk enamel elements. In the enamel surface, F and Cl co-varied. In addition, Cl was an element often found to co-vary with other enamel surface elements. PMID- 11063018 TI - Atomic force microscopy studies of crystal surface topology during enamel development. AB - During the secretory stage of enamel development, the hydroxyapatite crystals appear as thin ribbons which grow substantially in width and thickness during the later maturation stage. In this study, the atomic force microscope (AFM) was used to investigate developmentally-related changes in deproteinized enamel crystal surface topography in normal animals and in those receiving daily doses of fluoride. The AFM revealed previously undescribed surfaces features, some of which may represent growth sites or different crystalline phases. Secretory stage crystals had greater surface rugosity and were more irregular, with spherical sub structures of 20-30 nm diameter arranged along the "c"-axis. Maturation stage crystals were smoother and larger but revealed both subnanometer steps and lateral grooves running parallel to the "c"-axis. Crystals from fluorotic tissue showed similar features but were more irregular with a higher degree of surface roughness, suggesting abnormal growth. The AFM may prove an important adjunct in determination of the mechanisms controlling crystal size and morphology in skeletal tissues. PMID- 11063019 TI - Subunit compartments of secretory stage enamel matrix. AB - Three quarters of the micro-environment of early secretory stage enamel consist of protein and water. The physical arrangement of this enamel matrix is closely related to enamel crystal growth and habit. In the present study, structural components of developing enamel were analyzed using atomic force microscopy, transmission electron microscopy, immuno-ultracryotomy, and electron diffraction. Atomic force images revealed spherical subunits measuring between 108 nm and 124 nm in diameter. Transmission electron micrographs indicated that developing crystals were surrounded by an electron dense coat which may be rich in proteins. Transmission electron micrographs and electron diffraction studies supported a concept in which initial enamel crystals consist of amorphous calcium phosphate and later fuse to hydroxyapatite. Cryo-immuno electron microscopy demonstrated homogeneous distribution of amelogenin epitopes within the entire enamel matrix. The current study suggests an intricate role of protein aggregation phenomena involved in initial enamel crystal growth and habit. PMID- 11063020 TI - Helix structure of ribbon-like crystals in bovine enamel. AB - In the early mineralized enamel crystals, ribbon-like crystals appear near the ameloblasts. Some ribbon-like crystals showed helical or spiral structure within restricted environment during the preparation of embryonic bovine specimens for electron microscope. These specimens did not suffer from the cutting damages nor staining effects. The main cause of the helix structure is considered a result of the dehydration during preparation. The periodic structure must reflect the regularity of initial enamel crystals. If dehydration caused the ribbon-like crystal to induce the periodic helix, it is one possibility that the earliest enamel crystal is OCP which has been proposed as a precursor of HA. Because it is considered that OCP is more sensitive to dehydration and more symmetric structure than biological HA. The periodicity of the helical ribbon-like structure was about 25 to 55 nm long and could be compared to the periodicity of organic helices which had observed in an immature rat enamel. PMID- 11063021 TI - A study on the promotion and suppression of demineralization of human dental hard tissues and hydroxyapatite. AB - Demineralization of human dental enamel and dentine and their analogue compound, hydroxyapatite, was examined by using pH-metry to measure the time-courses of neutralization of acetate, formate, lactate or propionate buffer solution or of acidification of EDTA solution. The extent of neutralization by enamel, dentine and hydroxyapatite was different for each acid but increased in the same order: propionate, acetate, lactate and formate. This order was consistent with that of the K values of these acids. The pH-metry was used to determine the influences of sodium chloride and sucrose on demineralization of enamel, dentine and hydroxyapatite by acetate, formate, lactate and propionate and by EDTA. The demineralization by these bioorganic acids was suppressed by sucrose but promoted by sodium chloride, except that the demineralization of enamel by acetate and propionate was little affected. The demineralization of enamel, dentine and hydroxyapatite by EDTA was little affected by sucrose but promoted by sodium chloride. The promotive effect of sodium chloride on demineralization may be due to the increasing of solubility product by this salt and the suppressive effect of sucrose may be due to the formation of a calcium saccharate formed from the sucrose reacted with calcium on the surface of apatite crystal and/or the reduction of solubility product by the sucrose. In this study, it was also ascertained that the use of pH-metry made it possible to determine easily the demineralization. PMID- 11063022 TI - Enamel mineralization and an initial crystalline phase. AB - In this communication, we summarized our recent experimental approaches to an unsettled issue, i.e., the nature and role of an acidic precursor in enamel mineralization. The objectives we specially focused our attention on are: the composition, structure and high resolution images of enamel crystals at various developmental stages, thermodynamic and kinetic consideration of octacalcium phosphate (OCP) vs hydroxyapatite (HA) precipitation in physiological media simulating the enamel fluid, reversible changes in the composition and structure of OCP, effects of fluoride at low concentrations and enamel proteins on OCP hydrolysis, and adsorption of enamel proteins onto OCP and fluoridated hydrolysates at neutral pH and room temperature. On the basis of all experimental evidence, we propose that enamel crystal growth comprises two events: the two dimensional growth of an OCP-like precursor in a narrow outermost zone adjacent to the ameloblasts and the subsequent overgrowth of apatite units on the template under discrete fluid environment in the underlying region distant from the cell layer. The experimental data also support the concept that the whole process of enamel mineralization is modulated substantially through interaction between enamel proteins and crystals including the acidic precursor. PMID- 11063023 TI - General principle of ordered apatitic crystal formation in enamel and collagen rich hard tissues. AB - The biomineralization processes in different hard tissues like enamel, circumpulpal dentine, epiphyseal growth plates were analyzed morphologically and ultrastructurally by an energy filtering transmission electron microscope. In the primary stage of crystal formation Ca- and phosphate-ions accumulate at charged sites, "active sites", along the fiber matrix-molecules of the extracellular matrix. After exceeding the critical radius for nucleation, crystal nuclei appear that develop to "chains" of stable nanometer-sized paracrystalline particles. In the latest studies of small area electron diffraction it was found that in the earliest stage of crystal formation these mineral chains show a parallel orientation in the direction of the c-axis of apatite. This was supported by a texture of the 002 reflection in the corresponding diffraction patterns. Since apatite is bipolar in this direction crystal growth would be in like manner in both directions. Thus the center-to-center distances between nucleating sites along the matrix macromolecules show with the chains of nanometer islands the same process of biomineralization in the different mineralizing hard tissue systems. This way of crystal formation might be a general principle of apatitic biomineralization. PMID- 11063024 TI - Fragmentation of the distal portion of Tomes' processes of secretory ameloblasts in the forming enamel of rat incisors. AB - In order to investigate enamel and dentin phospholipid metabolic pathways, two separate experiments were carried out. Firstly, rats were given chloroquine, a drug which induces a lipidosis-like disease. Extensive accumulation inside lysosomes was seen in all the groups of cells in the forming part of the rat incisor, except secretory ameloblasts which were unaffected by the drug. Secondly, the uptake and fate of 3H-choline were studied by radioautography on rats fed either normally or on an essential fatty acid deficient diet (EFAD). Four hours after the injection of the precursor, incorporation reached a maximum then decreased gradually. At 4 days the forming enamel displayed higher silver grain density than any other compartment. In EFAD rats 3H-choline incorporation was decreased drastically in each compartment except in the forming enamel which was not affected by the deficiency. The longer retention of the labeling in the forming enamel and the lack of lysosomal accumulation in chloroquine-treated secretory ameloblasts support the hypothesis that fragments of the distal Tomes' process are released during enamel formation. Disconnected from the cells, membrane remnants are neither reinternalized nor subjected to further degradation inside lysosomes. PMID- 11063025 TI - Effect of some physico-chemical properties of matrix on lengthwise and oriented growth of octacalcium phosphate crystal. AB - Relationship between some physicochemical properties of matrix, such as viscosity, density or structure of the framework, and crystal growth of octacalcium phosphate (OCP) was studied using various concentrations of polyacrylamide gels at 37 degrees C and at pH 6.5. Reaction was carried out in a model system of enamel formation, where calcium solution (30 mM) and phosphate solution (5 mM) were separated by a cation selective membrane with polyacrylamide gel on the PO4 side. OCP grew in ribbon-like morphology in 5-20% polyacrylamide gels. Crystal size of OCP decreased with an increase in gel concentration. In 30% gel, growth of OCP was disturbed, In 7.5% polyacrylamide gel, 1% albumin reduced the crystal size of OCP. In contrast, 1% enamel proteins did not much reduce the crystal size of OCP and OCP crystallized in its characteristic ribbon-like morphology. PMID- 11063026 TI - pH and carbonate levels in developing enamel. AB - Our earlier studies showed that the surface of developing and calcifying enamel changes its pH alternatively along the tooth axis when stained with pH indicating dyes. Based on the pH conditions, the enamel at this stage was distinguished as neutral zone (N1 and N2) and acid zone (A1 and A2). The aim of the present study was to correlate changes of pH with proteolytic activity and crystal size of the calcifying bovine enamel. Specimens of developing bovine enamel were separated into four maturing stages using pH staining methods. Crystal chemistry of the developing enamel was investigated using thermogravimetry (TGA), ICP emission spectrometry, X-ray diffractometry (XRD) and infrared spectroscopy (IR). Previous biochemical analysis of proteolytic enzyme activity from enamel indicated that the optimal pH of the major protease was approximately pH 6.0, coinciding with the pH of the A1 zone. IR, TGA and XRD analyses showed that most of the organic components of the enamel decomposed at 580 degrees C. Higher levels of carbonate were observed in the secretory stages than in mature enamel. The Ca/P molar ratio of the enamel apatite was lower than the stoichiometric value of 1.67. These results suggest that growth and maturation of enamel apatite crystals is related to a decrease in the carbonate level and appear to be related to the alternative calcification and decomposition of enamel proteins. PMID- 11063027 TI - Two new in vitro calcification systems showing the higher calcifiability of enamel proteins than dentin and bone matrices. AB - One of the difficulties in simulating in vivo calcification by in vitro experiments is how to prepare and apply a suitable calcifying solution. We have previously developed an entirely new model system consisting of 40% acrylamide gel blocks that contains matrix proteins and is immersed in fetal calf serum at 37 degrees C. (40% gel system) for 18 hr (Connect. Tissue Res., 33, 185, 1995). The gels were analyzed for immobilized calcium. In this system bovine enamel proteins (0.1% in the gel) showed the highest calcifiability among the tested matrices, followed by insoluble bovine dentin, bone and skin collagens. The 40% gel system provides a barrier for high molecular weight inhibitor molecules in the body fluid. The new calcifying system developed in this study consists of the matrix protein sealed in dialysis tubing within a glass chromatography column that was eluted with a calcifying solution. In this system (dialysis tubing system), again the enamel protein showed higher calcifiability than dentin, bone and skin collagens. It was also shown that enamel proteins became not only a reversible opaque gel, but also a relatively-irreversible coagulant, if the solution contained calcium and phosphate ions at concentration below saturation (1 mM calcium and 1 mM phosphate). With both systems combined, deposition and crystal growth of minerals in enamel proteins will be better understood than with previous methods. PMID- 11063028 TI - Biochemical characterization of recombinant mouse amelogenins: protein quantitation, proton absorption, and relative affinity for enamel crystals. AB - Four recombinant mouse amelogenins, which varied by the presence or absence of the exon 4 encoded segment as well as the carboxyl-terminus were heterologously expressed and purified from bacteria. The rM193 and rM179 contain the carboxyl terminus, whereas the rM180 and rM166 do not. The rM193 and rM180 contain the polypeptide segment encoded by exon 4 of the amelogenin gene. A precisely weighed sample of purified rM179 was quantified by Lowry, Bicinchoninic Acid and Bradford assays. It was determined that these protein quantification methods characteristically under or overestimate the amount of amelogenin. The calculated correction factors were: Lowry (x 1.35), BCA (x 1.96), and Bradford (x 0.78). Recombinant mouse amelogenin (rM179) was characterized with respect to its hydrogen ion binding properties. The protein absorbs 11.9 +/- 1.7 protons during a pH change from 8.0 to 5.0, suggesting that amelogenins buffer the enamel fluid in vivo. Crystal binding experiments were performed using rM193, rM180, rM179 and rM166. The carboxyl-terminus enhanced the binding of amelogenin to enamel crystals while the exon 4 encoded segment did not appreciably affect crystal binding. PMID- 11063029 TI - The pH dependent amelogenin solubility and its biological significance. AB - Amelogenins are a group of extracellular enamel matrix proteins which are believed to be involved in the regulation of the size and habit of enamel crystals. The aim of this study was to compare the solubility properties of several amelogenins in various pH (4.0-9.0) solutions with an ionic strength (IS) of 0.15 M using the Micro BCA protein assay at 25 degrees C or 37 degrees C. The solubility of the recombinant amelogenin rM179 was lowest (0.7 mg/ml) close to its isoelectric point and it increased below and above this point. The solubility of the recombinant amelogenin rM166 remained almost the same (1-2 mg/ml) as the pH rose from 6.0 to 9.0 and it increased as the solution became more acidic. Synthetic "tyrosine-rich amelogenin polypeptide" (TRAP) was extremely insoluble (<0.2 mg/ml) in the pH range studied while synthetic "leucine-rich amelogenin polypeptide" (LRAP) was readily soluble (>3.3 mg/ml). The native porcine amelogenin with apparent molecular weight 25 kDa shared similar solubility behavior to rM179. The porcine 23 kDa amelogenin was only sparingly soluble (0.3 0.8 mg/ml) over a wide range of pH. Interestingly, the porcine 20 kDa amelogenin was remarkably soluble in the pH range of 4.0 to 6.0 (approximately 12 mg/ml), but the solubility dropped strikingly to only approximately 0.2 mg/ml at pH larger than approximately 7.0. The strong dependence of amelogenin solubility on solution pH may be involved in the regulation of aggregation, enzymatic degradation and the binding properties of amelogenins, thus playing an important role in enamel biomineralization. PMID- 11063030 TI - Enamel specific protein kinases and state of phosphorylation of purified amelogenins. AB - Ameloblastic tissue samples from unerupted bone molars were used to prepare subcellular enamel protein kinase preparations, nuclear + plasma membrane, cytosolic and microsomal, and used in in vitro phosphorylation of purified 20 kDa bovine amelogenin in the presence of 32P-ATP. Both cytosolic and microsomal preparations can phosphorylate purified native amelogenins, the addition of Ca2+ slightly increased the microsomal enzyme activity or at least did not inhibit the activity, whereas the presence of Ca2+ substantially decreased the cytosolic kinase activity towards phosphorylation of amelogenins. A comparative analysis using the enamel microsomal kinase against osteopontin, dephosphorylated casein and bone sialoprotein showed no phosphorylation of the first two proteins, and only minor phosphorylation of the bone sialoprotein. Overall, the present work demonstrates for the first time that the protein kinase responsible for the phosphorylation of amelogenins is a novel kinase, which is not inhibited by Ca2+, unlike the microsomal protein kinase (casein kinase type-II) of bone which phosphorylates secretory proteins osteopontin and bone sialoprotein and is strongly CaZ+ inhibited. The direct phosphoserine analysis on the purified bovine 20 kDa amelogenin indicated the presence of 0.8 moles of phosphoserine/mole protein naturally occurring, consistent with the quantitative analysis of 14C radiolabeling of phosphoserines by conversion to dehydroalanine and in situ reaction with the thiol agent, 14C-mercaptoethanol, 0.64 moles 14C incorporated/mole 20 kDa amelogenin. The purified low Mramelogenins 5.3 kDa E4 (TRAP) and 7.2 kDa E3 (LRAP), were also derivatized by 14C-mercaptoethanol, providing 0.46 and 0.88 moles 14C-incorporated/mole respectively. Further studies of the 14C-radiolabeled E4 amelogenin by sequence analysis confirmed one site of label to be at position 16 from the N-terminal and hence provided a direct evidence for the naturally occurring phosphoserine residue at this position. PMID- 11063031 TI - Does amelogenin nanosphere assembly proceed through intermediary-sized structures? AB - We have proposed that these nanosphere structures are functionally involved in the organization and control of initial enamel biomineralization at the ultrastructural level. Based on the observed nanosphere hydrodynamic radii (18-20 nm diameter) computation suggests these structures to be compounded of some 100 amelogenin monomers, raising the question as to the possible molecular mechanism for the assembly of such structures? Based on recent dynamic light scattering experiments using the recombinant murine amelogenin M179, and employing a newer size distribution algorithm we now report that the size distribution data for M179 are better described by a bimodal distribution model, than the monomodal distribution as previously described. We suggest that amelogenin nanosphere assembly proceeds through intermediate structures (perhaps represented in vivo by "stippled material") of some 4-5 nm hydrodynamic radius, and computed to comprise 4-6 amelogenin monomers. We suggest that such intermediary, sub-unit structures, assemble through inter-molecular hydrophobic interactions to generate the 20 nm diameter nanospheres observed by TEM in the secretory stage enamel matrix. PMID- 11063032 TI - Identification of rat enamel organ RNA transcripts using differential-display. AB - Enamel formation is a complex process which involves the expression of a number of genes, the most obvious being those related to the mineralized extracellular matrix. In this study the differential-display technique, first described by Liang and Pardee, has been used to identify genes specifically expressed in enamel organ cells. By comparing results obtained from RNA derived from rat enamel organ with RNA derived from other cellular sources, a number of differentially expressed transcripts have been identified. The nucleotide sequences of two of these have been analyzed and shown to have no homology with any previously published sequences. Further analysis will provide information on the type of protein that they may encode, their tissue distribution and their potential role in enamel formation. PMID- 11063033 TI - Identification of tuftelin- and amelogenin-interacting proteins using the yeast two-hybrid system. AB - Biomineralization of enamel is a complex process that involves the eventual replacement of an extracellular protein matrix by hydroxyapatite crystallites. To date four different enamel matrix proteins have been identified; the amelogenins, tuftelin, enamelin and ameloblastin. Assembly of the enamel extracellular matrix from these component proteins is believed to be critical in producing a matrix competent to undergo mineral replacement. Enamel formation is a complex process and additional proteins are likely to have a role in the assembly of the extracellular matrix. In order to identify additional proteins involved in the assembly process, the yeast two-hybrid system developed by Fields and Song (1989) has been implemented. This system allows for the identification of unknown proteins that interact with proteins of interest. Typically a known protein is used as "bait" to screen a cDNA expression library of interest. In our studies, tuftelin or amelogenin have been used to screen a mouse tooth library produced from one day old pups. A library screening of six million clones with amelogenin as bait resulted in eleven positive clones all of which show high homology to the human leukocyte antigen-B (HLA-B) associated transcript (BAT) family of genes. A library screening of one million clones using tuftelin as the bait identified twenty-one tuftelin-interacting proteins. Ten of these proteins are either keratin K5 or keratin K6, four are constitutively expressed and the remaining seven are novel. Further characterization of the proteins shown to interact with amelogenin or tuftelin may shed additional light on this complex process of enamel matrix assembly. PMID- 11063034 TI - Induction of amelogenin and ameloblastin by insulin and insulin-like growth factors (IGF-I and IGF-II) during embryonic mouse tooth development in vitro. AB - Insulin and insulin-like growth factors (IGF-I and IGF-II) are considered pleiotropic, acting as both mitogen and differentiation factors. Several investigators have demonstrated the expression of insulin, IGFs, their cognate receptors and IGF binding proteins during tooth morphogenesis. Previous work done in our laboratory indicated that exogenous insulin and IGFs induce the accumulation of enamel extracellular matrix on mouse mandibular molars cultured in a serumless, chemically defined medium. In order to determine the level of control of these factors in the induction of enamel biomineralization, we designed experiments to quantitate mRNAs for enamel specific-gene products. Mandibular first molars (MI) obtained from E15 Swiss Webster mice were placed in organ culture in the presence of insulin (1,000 ng/ml), IGF-I (100 ng/ml) or IGF II (100 ng/ml) for 6, 12 and 18-days. At termination date, the RNA was extracted and the concentration of mRNAs for amelogenin, tuftelin and ameloblastin were determined using a quantitative competitive reverse transcription-polymerase chain reaction (RT-PCR) technique (PCR mimic). Our results showed that after 6 days in culture; treatment with insulin, IGF-I and IGF-II increased the synthesis of amelogenin and ameloblastin. In contrast, the expression of tuftelin mRNA was not affected by either factor. In conclusion, our studies showed that the increase in enamel matrix formation by overexpression of IGFs is the result of transcriptional regulation of enamel specific proteins like amelogenin and ameloblastin but not tuftelin. These studies also suggest that the regulatory mechanisms controlling tuftelin gene expression are different than the mechanisms regulating ameloblastin and amelogenin transcription. PMID- 11063035 TI - Transgene animal model for protein expression and accumulation into forming enamel. AB - Understanding the cellular and molecular events that regulate the formation of enamel is a major driving force in efforts to characterize critical events during amelogenesis. It is anticipated that through such an understanding, improvements in prevention, diagnosis and treatment-intervention into heritable and acquired diseases of enamel could be achieved. While knowledge of the precise role of an enamel-specific protein in directing the formation of inorganic crystallites remains refractory, progress has been made with other aspects of amelogenesis that can be brought to bear on the subject. One such area of progress has been with the identification of an ameloblast-lineage specific amelogenin gene promoter. This promoter can be used to direct the expression of enamel-specific proteins, as well as the expression of proteins foreign to amelogenesis, into the enamel extracellular matrix where their effect on biomineralization can be ascertained in a prospective manner. The resulting enamel from such animals can be examined by morphologic and biochemical modalities in order to identify the effect of the transgene protein on enamel crystallite formation and subsequent biomineralization. This manuscript outlines such a strategy with the potential for enhancing our understanding of amelogenesis. PMID- 11063036 TI - Cyclic AMP-receptor proteins in the enamel matrix. AB - Cyclic AMP receptor proteins (cARP) are present in a variety of cell types. Intracellularly, they are the regulatory (R) subunits of type II cyclic AMP dependent protein kinase (PKA: E.C.2.7.1.37). Additionally, cARP are secretory products of several cell types.[1] That cARP are present in and secreted by ameloblasts into the enamel matrix of the rat incisor was demonstrated by photoaffinity labeling, Western blotting and immunogold cytochemistry. Gold particles were present over cytoplasmic regions including Tomes' Processes of secretory ameloblasts, secretory granules and in the Golgi region. Specific RII labeling was seen in the enamel matrix, but not in dentin. The enamel matrix was more reactive during early maturation compared to the secretory stage of amelogenesis. Nuclear labeling with the RII antibody showed higher intensity in maturation than in secretory ameloblasts. These results demonstrate that cARP are expressed in ameloblasts and secreted into the enamel matrix. The role(s) of cARP in enamel matrix mineralization and the involvement of PKA-regulated pathways in enamel protein synthesis and secretion remain to be determined. PMID- 11063037 TI - Adolescents: at increased risk for osteoporosis? AB - Perhaps the most important factor in the primary prevention of osteoporosis is the attainment of an optimal peak bone during adolescence. In addition to endogenous factors, such as genetic and ethnic background, environmental factors such as dietary habits, physical activity, and sex hormone therapy, influence the accretion of bone mass during this critical period of skeletal growth. First, calcium dietary intake in adolescents is generally well less than the current recommended RDA of 1200 mg/day. Multiple studies of children and adolescents have demonstrated increases in bone mass with dietary calcium supplementation. Second, regarding physical activity, the overall impression is that a moderate amount of particularly weight-bearing exercise has a positive impact on bone. There appears, however, to be a threshold of intensity of physical activity over which a negative impact on bone occurs, particularly when the exercise is of an anaerobic nature or occurring in very thin, amenorrheic participants. Third, previous research suggests that the various forms of hormonal contraception exert differing effects on bone mass in adolescents, with levonorgestrel implants and combined oral contraceptives may be associated with a more positive effect on bone mass compared with that observed with depot medroxyprogesterone acetate. From a clinical perspective, approaches to optimizing peak bone mass in adolescents would include increasing calcium intake, whether in the form of dairy products, fortified foods, or supplements as well as encouraging participation at a moderate level, in weight-bearing exercise. Last, in adolescents with extensive risk factors and predicted long duration of use, subdermal implants or combined oral contraceptives may be the optimal hormonal methods of birth control. PMID- 11063038 TI - Withholding artificially provided nutrition and hydration from disabled children- assessing their quality of life. AB - This article focuses on quality of life determinations and limitation of treatment decisions for children with physical and mental disabilities. Issues are addressed through one pediatric convalescent center's ethical dilemma, deliberations and process for decision-making when the organization's definition of quality of life differed from that of the parents wishing to place their child there. The Ethics Committee suggested revised admission criteria to include provision of hydration and nutrition for future admissions. PMID- 11063039 TI - Problem identification in apparently well neonates: implications for early discharge. AB - The frequency, time of identification, and type of problems of newborns in an urban indigent population were prospectively studied during their hospital stay to evaluate feasibility of early hospital discharge. Eight percent (563) of 7,021 term and near-term low-risk infants developed one or more predefined problems. Of those with problems, 42.1% received therapy and/or a higher level of care. Tachypnea, temperature instability, and cyanotic episodes were the most frequently treated problems. Nearly 69% of all problems were detected after the initial examination, and 31% developed problems after 24 hours of age; 5% were transferred to the NICU. Problems occurring after 24 hours of age emphasize the need for follow-up within days after hospital discharge in this population. PMID- 11063040 TI - Physician reimbursement and the medical encounter: an observational study in Dutch pediatrics. AB - Pediatrician reimbursement is shifting from fee-for-service to a fixed salary. In the Netherlands, as physicians working on a fee-for-service basis have a financial interest in talking less and in carrying out more diagnostic tests and investigations, it may be questioned whether this will influence the structure and content of medical visits. With use of 302 videotaped outpatient encounters with either salaried or fee-for-service pediatricians, differences were examined in visit length, number of requests for diagnostic tests and investigations (laboratory test, endoscopy, and radiography), pediatrician-parent communication behaviors, and patient satisfaction. This investigation was carried out by means of bivariate and multilevel analysis. The results showed that the visits with salaried pediatricians lasted almost 4 minutes longer. This surplus time was not spent on social talk or on a more elaborate history taking but was used to provide more information and advice. In addition, salaried pediatricians engaged in more empathic behavior toward the patient, thereby facilitating a therapeutic relationship. No differences were found in the number of diagnostic tests and investigations or in patient satisfaction. It may be concluded that history taking and social talk took place in a fixed part of the visit. Salaried pediatricians spent more time on exchanging information with their patients and paid more attention to patient concerns and emotions. As the reimbursement shift is not likely to diminish the number of diagnostic tests and investigations and will increase the length of the medical visits, overall financial benefits may be limited. PMID- 11063041 TI - Clinical approach to fecal soiling in children. AB - Fecal soiling is common in childhood and can be caused by stool toileting refusal, fecal incontinence due to organic disease, or encopresis due to functional constipation. Anatomical, neurologic, and inflammatory causes for fecal soiling are ruled out by history and physical examination and, if necessary, by anorectal manometry, barium enema, and rectal biopsy. The initial treatment suggestion for children with stool toileting refusal is to put the child back into pull-ups or diapers. Most children with fecal soiling due to organic disease continue with some degree of incontinence despite optimal medical management. Antegrade enema administration helps those with severe fecal incontinence due to organic causes who do not respond to medical management. Successful treatment of constipation and encopresis requires a combination of medical therapy, nutritional intervention, behavioral intervention, and long-term compliance with laxative use. The combined treatment approach improves the constipation and encopresis in all patients who comply with the treatment program. In some children, cow's milk protein intolerance may be the cause. In them, cow's milk protein needs to be eliminated. PMID- 11063042 TI - Septal hematoma and abscess after nasal trauma. PMID- 11063043 TI - Cerebellar hemorrhage in preterm infants with intraventricular hemorrhage: a missed diagnosis? A patient report. PMID- 11063044 TI - The status of school health in pediatric residency training. PMID- 11063045 TI - Charge association and infantile hypertrophic pyloric stenosis. PMID- 11063046 TI - Combination therapy in mild to moderate bronchiolitis. PMID- 11063047 TI - Pica and intoxication in childhood. PMID- 11063048 TI - Suprapubic bladder aspiration with or without ultrasound guidance. PMID- 11063049 TI - Apoptosis: overview and signal transduction pathways. AB - Apoptosis is a form of cell death that is driven by an intrinsic cellular suicide program. The roles of apoptosis and other forms of programmed cell death in neural development, maintenance, and disease states are increasingly being recognized and defined. Therapies directed at the apoptotic program have seen at least some degree of success in animal models of neurodegenerative disease, vascular disease, and traumatic CNS injury. This article describes the signal transduction pathways that mediate apoptosis. Broadly speaking, intrinsic and extrinsic pathways for apoptosis activation may be distinguished, as can be cross talk between these two. These pathways converge on a system of proteases referred to as "capases" (cysteinyl aspartic proteinases), and modulators exist that multimerize, activate, amplify, or inhibit caspases. Activated caspases are the executioners of the apoptotic program, and carry out this function by cleaving specific cellular substrates. Modulation of this process holds promise as a therapeutic approach in neurotrauma. PMID- 11063050 TI - Caspase pathways, neuronal apoptosis, and CNS injury. AB - Caspases are a family of mammalian proteases related to the ced-3 gene of Caenorhabditis elegans. They mediate many of the morphological and biochemical features of apoptosis, including structural dismantling of cell bodies and nuclei, fragmentation of genomic DNA, destruction of regulatory proteins, and propagation of other pro-apoptotic molecules. Based on their substrate specificities and DNA sequence homologies, the 14 currently identified caspases may be divided into three groups: apoptotic initiators, apoptotic executioners, and inflammatory mediators. Caspases are activated through two principal pathways, known as the "extrinsic pathway," which is initiated by cell surface death receptor ligation, and the intrinsic pathway, which arises from mitochondria. Endogenous inhibitors, such as the inhibitors of apoptosis (IAP) family, modulate caspase activity at various points within these pathways. Upon activation, caspases appear to play an important role in sequelae of traumatic brain injury, spinal cord injury, and cerebral ischemia. In addition, they may also play a role in mediating cell death in chronic neurodegenerative conditions such as Alzheimer's disease, Huntington's disease, and amyotrophic lateral sclerosis. This article reviews the current literature on the role of caspases in acute and chronic CNS injury, and provides evidence for the potential therapeutic use of caspase inhibitors in the setting of these conditions. PMID- 11063051 TI - Bcl-2 family gene products in cerebral ischemia and traumatic brain injury. AB - The proto-oncogene bcl-2 plays a key role in regulating programmed cell death in neurons. The present review discusses the mechanisms by which bcl-2 family genes regulate programmed cell death, and their role in controlling cell death in cerebral ischemia and traumatic brain. Expression of several bcl-2 family members is altered in brain tissues after ischemia and trauma, suggesting that bcl-2 family genes could play a role in determining the fate of injured neurons. Furthermore, alteration of expression of bcl-2 family genes using transgenic approaches, viral vectors, or anti-sense oligonucleotides modifies neuronal cell death and neurological outcome after injury. These data suggest that the activity of bcl-2 family gene products participates in determining cellular and neurologic outcomes in ischemia and trauma. Strategies that either mimic the death suppressor effects or inhibit the death-promoter effects of bcl-2 family gene products may improve outcome after ischemia and trauma. PMID- 11063052 TI - Mitochondrial participation in ischemic and traumatic neural cell death. AB - Mitochondria play critical roles in cerebral energy metabolism and in the regulation of cellular Ca2+ homeostasis. They are also the primary intracellular source of reactive oxygen species, due to the tremendous number of oxidation reduction reactions and the massive utilization of O2 that occur there. Metabolic trafficking among cells is also highly dependent upon normal, well-controlled mitochondrial activities. Alterations of any of these functions can cause cell death directly or precipitate death indirectly by compromising the ability of cells to withstand stressful stimuli. Abnormal accumulation of Ca2+ by mitochondria in response to exposure of neurons to excitotoxic levels of excitatory neurotransmitters, for example, glutamate, is a primary mediator of mitochondrial dysfunction and delayed cell death. Excitoxicity, along with inflammatory reactions, mechanical stress, and altered trophic signal transduction, all likely contribute to mitochondrial damage observed during the evolution of traumatic brain injury. The release of apoptogenic proteins from mitochondria into the cytosol serves as a primary mechanism responsible for inducing apoptosis, a form of cell death that contributes significantly to neurologic impairment following neurotrauma. Although several signals for the release of mitochondrial cell death proteins have been identified, the mechanisms by which these signals increase the permeability of the mitochondrial outer membrane to apoptogenic proteins is controversial. Elucidation of the precise biochemical mechanisms responsible for mitochondrial dysfunction during neurotrauma and the roles that mitochondria play in both necrotic and apoptotic cell death should provide new molecular targets for neuroprotective interventions. PMID- 11063053 TI - Neuronal apoptosis after CNS injury: the roles of glutamate and calcium. AB - While a role has been well established for excitotoxic necrosis in the pathogenesis of traumatic or ischemic damage to the CNS, accumulating evidence now suggests that apoptosis may also be a prominent contributor. In this review we focus on the role of glutamate and attendant intracellular calcium influx in triggering or modifying excitotoxic necrosis and apoptosis, raising the possibility that calcium influx may affect these two death pathways in opposite directions. Incorporating consideration of both pathways will probably be needed to develop the most effective neuroprotective treatments for CNS injury. PMID- 11063054 TI - Free radical pathways in CNS injury. AB - Free radicals are highly reactive molecules implicated in the pathology of traumatic brain injury and cerebral ischemia, through a mechanism known as oxidative stress. After brain injury, reactive oxygen and reactive nitrogen species may be generated through several different cellular pathways, including calcium activation of phospholipases, nitric oxide synthase, xanthine oxidase, the Fenton and Haber-Weiss reactions, by inflammatory cells. If cellular defense systems are weakened, increased production of free radicals will lead to oxidation of lipids, proteins, and nucleic acids, which may alter cellular function in a critical way. The study of each of these pathways may be complex and laborious since free radicals are extremely short-lived. Recently, genetic manipulation of wild-type animals has yielded species that over- or under-express genes such as, copper-zinc superoxide dismutase, manganese superoxide dismutase, nitric oxide synthase, and the Bcl-2 protein. The introduction of the species has improved the understanding of oxidative stress. We conclude here that substantial experimental data links oxidative stress with other pathogenic mechanisms such as excitotoxicity, calcium overload, mitochondrial cytochrome c release, caspase activation, and apoptosis in central nervous system (CNS) trauma and ischemia, and that utilization of genetically manipulated animals offers a unique possibility to elucidate the role of free radicals in CNS injury in a molecular fashion. PMID- 11063055 TI - Role of ceramide in neuronal cell death and differentiation. AB - Ceramide is a sphingolipid metabolite that has been implicated in cellular apoptosis and differentiation. It has been shown to induce apoptosis in various mammalian cell lines and, more recently, has been implicated in neuronal apoptosis. Although the mechanisms of ceramide-induced cell death have not been fully elucidated, they appear to involve a number of signal transduction pathways, including proline-directed kinases, phosphatases, phospholipases, transcription factors, and caspases. Interestingly, ceramide also appears to promote survival and differentiation in certain neuronal systems, when applied at lower concentrations and/or at different developmental stages. Together, studies to date indicate an important multipotential role for this lipid in cell death and differentiation. PMID- 11063056 TI - Apoptosis after experimental stroke: fact or fashion? AB - This review examines the appearance of hallmarks of apoptosis following experimental stroke. The reviewed literature leaves no doubt that ischemic cell death in the brain is active, that is, requires energy; is gene directed, that is, requires new gene expression; and is capase-mediated, that is, uses apoptotic proteolytic machinery. However, sufficient differences to both classical necrosis and apoptosis exist which prevent easy mechanistic classification. It is concluded that ischemic cell death in the brain is neither necrosis nor apoptosis but is a chimera which appears on a continuum that has apoptosis and necrosis at the poles. The position on this continuum could be modulated by the intensity of the ischemic injury, the consequent availability of ATP and new protein synthesis, and both the age and context of the neuron in question. Thus the ischemic neuron may look necrotic but have actively died in an energy dependent manner with new gene expression and destruction via the apoptotic proteolytic machinery. PMID- 11063057 TI - Review of current evidence for apoptosis after spinal cord injury. AB - The initial mechanical tissue disruption of spinal cord injury (SCI) is followed by a period of secondary injury that increases the size of the lesion. The secondary injury has long been thought to be due to the continuation of cellular destruction through necrotic (or passive) cell death. Recent evidence from brain injury and ischemia suggested that cellular apoptosis, an active form of programmed cell death seen during development, could play a role in CNS injury in adulthood. Here, we review the evidence that apoptosis may be important in the pathophysiology of SCI. There is now strong morphological and biochemical evidence from a number of laboratories demonstrating the presence of apoptosis after SCI. Apoptosis occurs in populations of neurons, oligodendrocytes, microglia, and, perhaps, astrocytes. The death of oligodendrocytes in white matter tracts continues for many weeks after injury and may contribute to post injury demyelination. The mediators of apoptosis after SCI are not well understood, but there is a close relationship between microglia and dying oligodendrocytes, suggesting that microglial activation may be involved. There is also evidence for the activation of important intracellular pathways known to be involved in apoptosis in other cells and systems. For example, some members of the caspase family of cysteine proteases are activated after SCI. It appears that the evolution of the lesion after SCI involves both necrosis and apoptosis. It is likely that better understanding of apoptosis after SCI will lead to novel strategies for therapeutic interventions that can diminish secondary injury. PMID- 11063058 TI - Apoptosis after traumatic brain injury. AB - Apoptosis of neurons and glia contribute to the overall pathology of traumatic brain injury (TBI) in both humans and animals. In both head-injured humans and following experimental brain injury, apoptotic cells have been observed alongside degenerating cells exhibiting classic necrotic morphology. Neurons undergoing apoptosis have been identified within contusions in the acute port-traumatic period, and in regions remote from the site of impact in the days and weeks after trauma. Apoptotic oligodendrocytes and astrocytes have been observed within injured white matter tracts. We review the regional and temporal patterns of apoptosis following TBI and the possible mechanisms underlying trauma-induced apoptosis. While excitatory amino acids, increases in intracellular calcium, and free radicals can all cause cells to undergo apoptosis, in vitro studies have determined that neural cells can undergo apoptosis via many other pathways. It is generally accepted that a shift in the balance between pro- and anti-apoptotic protein factors towards the expression of proteins that promote death may be one mechanism underlying apoptotic cell death. The effect of TBI on regional cellular patterns of expression of survival promoting-proteins such as Bcl-2, Bcl-xL, and extracellular signal regulated kinases, and death-inducing proteins such as Bax, c-Jun N-terminal kinase, tumor-suppressor gene, p53, and the caspase family of proteases are reviewed. Finally, in light of pharmacologic strategies that have been devised to reduce the extent of apoptotic cell death in animal models of TBI, our review also considers whether apoptosis may serve a protective role in the injured brain. PMID- 11063059 TI - An evaluation of the effectiveness of targeted social marketing to promote adolescent and young adult reproductive health in Cameroon. AB - This study examines the effectiveness of the PSI/PMSC Horizon Jeunes youth targeted social marketing program for improving adolescent reproductive health in urban Cameroon. The program targeted adolescents through peer education, youth clubs, mass media promotion, and behavior change communications. After about 13 months of intervention, knowledge of the program was nearly universal, and the majority of youths had direct contact with the program. Program effectiveness is examined using a quasi-experimental research design with a preintervention and postintervention survey in an intervention and comparison site. The intervention had a significant effect on several determinants of preventive behavior, including awareness of sexual risks, knowledge of birth control methods, and discussion of sexuality and contraceptives. The intervention increased the proportion of female youths who reported using oral contraceptives and condoms for birth control. However, condom use is not yet consistent. Although the proportion of young men who reported using condoms for birth control also increased, this change could not be attributed to the intervention. Although this short intervention successfully increased the reported use of various birth control methods, including condoms, there is no evidence that the intervention increased use of condoms for STD prevention of sexually transmitted diseases. PMID- 11063060 TI - Age-related risk for HIV infection in men who have sex with men: examination of behavioral, relationship, and serostatus variables. AB - The study examined behavioral, relationship, and serostatus variables that potentially contribute to HIV infection risk in three age groups of men who have sex with men (MSM). MSM recruited in West Hollywood, California self-administered a questionnaire measuring unprotected insertive anal intercourse (UIAI) and unprotected receptive anal intercourse (URAI) with primary and nonprimary partners. The following relationship/serostatus variables were also assessed: recency of HIV testing, knowledge of own HIV serostatus, perception of partner's serostatus, seroconcordance (self and partner seronegative), and self-reported monogamy status. The prevalence of UIAI and URAI was higher with primary than nonprimary partners. These sexual risk behaviors with primary partners were substantially more prevalent among men younger than 25 years of age than among men aged 25 to 30 or over age 30. UIAI and URAI with nonprimary partners were uncommon in each age group, and there were no significant age differences on the serostatus and relationship variables. The findings suggest that young MSM may be at elevated risk for contracting HIV by virtue of their sexual risk behavior with primary partners. Targeted interventions for MSM need to address sexual risk in the context of primary relationships. PMID- 11063061 TI - Psychosocial predictors of unprotected anal intercourse in a sample of HIV positive gay men who volunteer for a sexual risk reduction intervention. AB - This study used logistic regression analysis to identify psychosocial predictors of unprotected anal intercourse in a sample of 212 HIV positive gay men who were enrolled in a sexual risk reduction intervention. A combination of five psychosocial variables (i.e., age, avoidant coping, loneliness, depressive symptoms, and impulsivity) reliably discriminated between men who had engaged in unprotected anal intercourse (UAI) with HIV negative or serostatus unknown partners and those who had not engaged in this high risk behavior. Avoidant coping emerged as the best psychosocial predictor that distinguished between the two groups. Men who had UAI during the previous 4 months had significantly higher scores on avoidant coping as compared with the no-UAI group. These findings suggest the need to design intervention programs for HIV positive gay men that addresses coping strategies in the context of high-risk sexual behaviors. PMID- 11063062 TI - Screening for transmission behaviors among HIV-infected adults. AB - Existing data suggest that a substantial proportion of HIV seropositive adults reduce their substance use and sexual risk behaviors after learning their serostatus. Given the importance of implementing preventive interventions, a screening tool is desirable to allow for a brief method of identifying those who continue to engage in transmission acts. This report explores the sensitivity and specificity of a brief screener used to identify persons living with HIV who engage in unprotected sexual intercourse and/or injection drug use. HIV positive clients of a large AIDS service organization in Los Angeles (N = 178) were screened with a brief, 7-item questionnaire, and responses on the screening interview were compared with self-reports obtained in an in-depth computerized assessment administered on the same day. Participants were randomized to receive the in-depth computerized assessment administered by an interviewer, or a self administered audio-computer assisted interview. Screener sensitivity and specificity rates were relatively low (68% and 78%, respectively), and sensitivity was lower among African Americans than non-African Americans. Multivariate logistic regression analyses indicated that false negative screener responses were associated with recent sexually transmitted disease experience, controlling for gender, ethnicity, and interview administration mode. Effective strategies need to be developed that screen persons living with HIV who may be transmitting HIV to their sexual or drug-injecting partners. PMID- 11063063 TI - Impact of HIV-positive speakers in a multicomponent, school-based HIV/STD prevention program for inner-city adolescents. AB - Qualitative and quantitative data from Safer Choices, a school-based multicomponent HIV prevention program, were examined to determine the impact of HIV-positive speakers on inner-city adolescents' HIV risk perception and empathy for people with HIV or AIDS. Inductive analyses were used to assess student reactions to speakers. Multilevel regression modeling techniques were used to analyze student survey data (n = 1,491) to determine the effect of speakers alone, as well as in combination with the multicomponent intervention, and a knowledge-based curriculum (comparison condition). Results showed that speakers were highly popular with students and teachers, and had a positive short-term impact on students' attitudes. Although not statistically significant, the combination of intervention and speakers had the greatest impact on outcome variables. Integrating HIV-positive speakers into multicomponent programs may have a positive impact on inner-city youth. Utilizing speakers without other educational components may have minimal effects. Strategies for training and utilizing HIV-positive speakers in school settings are included. PMID- 11063064 TI - Do characteristics of HIV/AIDS education and training affect perceived training quality? Lessons from the evaluation of seven projects. AB - Initial and continuing HIV/AIDS education and training has been a critical way to bring the nation's health providers up to date on emerging developments and approaches. This study reports cross-cutting findings from seven HIV/AIDS education and training projects. Trainers described over 600 training sessions from these projects in terms of their structural characteristics and design elements, while trainees described these sessions on several dimensions related to training quality. Training characteristics were compared to trainee assessments of training quality. Using a decision-tree analytic approach for major training attributes, considerable support emerged for links between training characteristics and perceived quality of the HIV/AIDS training experience. More favorable quality ratings were associated with certain projects, the training setting, the types of trainees served by the training, the intended training impact, discussion of special populations, and training methods involving interactive learning. With increased knowledge regarding how these educational experiences relate to the ways they are perceived and processed, more targeted approaches to training design on HIV/AIDS can be developed. PMID- 11063065 TI - Supporting the transfer of HIV prevention behavioral research to public health practice. PMID- 11063066 TI - A technology transfer model for effective HIV/AIDS interventions: science and practice. AB - The widespread use of effective, science-based interventions to motivate and sustain behavior change provides an important approach to reducing the spread of HIV. The process of disseminating information about effective interventions and building capacity for implementing them in field settings must be improved, however. Starting with a review of diffusion of innovations and technology transfer literature, we offer a technology transfer model for HIV interventions. We identify participants and activities directed toward the use of effective interventions by prevention services providers (e.g., health departments and community-based organizations) in each phase of technology transfer: preimplementation, implementation, and maintenance and evolution. Preimplementation activities focus on selecting an intervention and preparing for implementation. Implementation activities include initial implementation and process evaluation. Maintenance and evolution are ongoing with continued support for and evaluation of the intervention. This article takes the perspective of providers. Other perspectives are presented elsewhere in this issue. PMID- 11063067 TI - Strengthening HIV prevention: application of a research-to-practice framework. AB - As the HIV epidemic continues to affect at-risk and vulnerable populations, providers strive to improve prevention programs, in part by seeking new interventions with greater effects. Although interventions with scientific evidence of effectiveness are vital to this effort, many challenges limit access to research products. We examine key challenges and offer a framework for moving research to practice, one in which research steps are linked to practice steps and all these activities take place in a complex and dynamic environment. The Replicating Effective Programs (REP) project of the Centers for Disease Control and Prevention and other technology transfer activities illustrate the operation of this framework for HIV prevention. Further actions to improve technology transfer are called for. These include reducing time from study design to practice; learning from field-based implementations; providing guidance about fidelity to, and tailoring of, science-based interventions; improving linkages among consumers, providers, and researchers; and seeking additional resources. PMID- 11063068 TI - Replicating effective programs: HIV/AIDS prevention technology transfer. AB - The Centers for Disease Control and Prevention (CDC) works to prevent HIV infection in collaboration with community and state partners. CDC is identifying effective interventions from the research literature and disseminating these interventions to its prevention partners. This article presents the methods used by CDC scientists and original intervention researchers in CDC's Replicating Effective Programs (REP) project to (a) translate some HIV prevention behavioral intervention research into materials with enough detail and clarity that state and community partners can select and implement effective interventions and (b) transfer and support these technologies so that they can be implemented successfully. The experience of the REP project indicates that technology transfer is complex. Interventions need to be adapted to local circumstances. Prevention partners need written materials, training, and technical assistance. Researchers need to collaborate with prevention program providers to develop interventions that are feasible for prevention partners to conduct. PMID- 11063069 TI - Bridging research and practice: community-researcher partnerships for replicating effective interventions. AB - Long-term collaborations among researchers, staff and volunteers in community based agencies, staff in institutional settings, and health advocates present challenges. Each group has different missions, procedures, attributes, and rewards. This article reviews areas of potential conflict and suggests strategies for coping with these challenges. During the replication of five effective HIV prevention interventions, strategies for maintaining mutually beneficial collaborations included selecting agencies with infrastructures that could support research-based interventions; obtaining letters of understanding that clarified roles, responsibilities, and time frames; and setting training schedules with opportunities for observing, practicing, becoming invested in, and repeatedly implementing the intervention. The process of implementing interventions highlighted educating funders of research and public health services about (a) the costs of disseminating interventions, (b) the need for innovation to new modalities and theories for delivering effective interventions, and (c) adopting strategies of marketing research and quality engineering when designing interventions. PMID- 11063070 TI - From science to application: the development of an intervention package. AB - Many community-based organizations and health departments want to implement HIV prevention interventions with scientifically demonstrated effectiveness. The Replicating Effective Programs (REP) project supported researchers in developing intervention packages designed to help prevention partners replicate effective programs in their settings. Intervention packages convey the intervention's foundation, components, and methods and are one part of a larger system needed to transfer research-based HIV prevention technology to service providers. Implementation packages were developed using a multistage process. The original researchers drafted the materials, advisory groups reviewed the packages, and adopting agencies used the materials in trial runs. The advisory groups and adopting agencies recommended extensive use of examples, thorough explanations about the rationale for each intervention component, explicit representation of people of color in the materials, clear statements about the intended audience(s), and an easy-to-use and visually appealing format. Packages were revised based on these recommendations and the outcomes of the trial runs. PMID- 11063071 TI - Orientation and training: preparing agency administrators and staff to replicate an HIV prevention intervention. AB - Effective orientation and training are fundamental to the successful implementation of any intervention because they communicate the critical first impressions of the intervention and the skills needed to conduct it. When research-based HIV prevention interventions are translated into practice, issues arise that require adaptation and expansion of the basic functions of orientation and training. This article identifies some of these issues by drawing on the experience of researchers in the Replicating Effective Programs (REP) project. The purpose, structure, and instructional approach of the orientation and training for administrators, staff, and volunteers are described in depth for one project, with comparisons and additional examples from others. Based on these descriptions, critical issues for orientation and training for replication are presented. These include extending orientation and training to a broad audience within the adopting agency, allocating sufficient time to ensure understanding of the intervention, and planning for staff turnover. PMID- 11063072 TI - Transfer of research-based HIV prevention interventions to community service providers: fidelity and adaptation. AB - HIV prevention research interventions usually follow protocols with specific procedures. If a community-delivered intervention uses the same procedures with the same populations as those in the original research, the behavior change effects should be similar. However, community-based providers may not replicate an intervention exactly as it was conducted in the effectiveness study. Adaptation may be needed to better meet the needs of the clients, community, or organization. We propose that interventions can be defined in terms of core elements likely to be responsible for effectiveness. These core elements cannot be changed without fundamentally changing the intervention, whereas other characteristics may be modified without altering effectiveness. HIV prevention researchers and service providers can collaborate to develop interventions that not only are effective but can also be successfully implemented by service organizations. If researchers actively involve service providers and community members in intervention planning, technology transfer goals can be better achieved. PMID- 11063073 TI - The role of technical assistance in the replication of effective HIV interventions. AB - This article examines the role of technical assistance (TA) in supporting the replication of proven HIV interventions. A case study of the replication of the VOICES/VOCES intervention elucidates the level and types of TA provided to support new users through the adoption process. TA included help in garnering administrative support, identifying target audiences, recruiting groups for sessions, maintaining fidelity to the intervention's core elements, tailoring the intervention to meet clients' needs, strengthening staff members' facilitation skills, troubleshooting challenges, and devising strategies to sustain the intervention. Two to four hours per month of TA were provided to each agency adopting the intervention, at an estimated monthly cost of $206 to $412. Findings illustrate how TA supports replication by establishing a conversation between the researcher TA providers experienced with the intervention and new users. This communication helps preserve key program elements and contributes to ongoing refinement of the intervention. PMID- 11063074 TI - Evaluation and technical assistance for successful HIV program delivery. AB - Agencies that provide HIV prevention programs (including agencies using science based interventions) need to conduct evaluation to facilitate the transfer of effective interventions, provide ongoing assistance to intervention providers, document services provided, demonstrate effectiveness, and improve programs. The Centers for Disease Control and Prevention (CDC) assists health departments, community-based organizations, and other CDC grantees with evaluation by providing direct, customized technical assistance and written guidance. Together, these activities assist grantees in conducting evaluations that provide the information that grantees need as their programs develop. PMID- 11063075 TI - Future directions and emerging issues in technology transfer between HIV prevention researchers and community-based service providers. AB - The public health objective of preventing new HIV infections can be achieved only through effective information exchange among service providers, researchers, and policymakers. The potential for successful transfer of research-based HIV prevention technology to service providers will be enhanced if investigators take into account in the research planning stage how interventions will be used in the field, seek early input from community members and service providers, test variations of interventions that may increase their practicality in applied settings, and determine the cost and effectiveness of intervention delivery. Strategies are needed to ensure that the experiences of service providers help to inform the HIV prevention research agenda, improve service organization infrastructure and capacity development, and facilitate organizational networking so that providers can use new-generation HIV prevention interventions. Policies are needed to facilitate the development of intervention packages, training, and ongoing technical assistance for service providers in implementing effective HIV prevention interventions. PMID- 11063076 TI - Hypomelanosis of Ito: clinical syndrome or just phenotype? AB - The term hypomelanosis of Ito is applied to individuals with skin hypopigmentation along the lines of Blaschko. Even though originally described as a purely cutaneous disease, subsequent reports have included a 33% to 94% association with multiple extracutaneous manifestations mostly of the central nervous and musculoskeletal systems leading to frequent characterization as a neurocutaneous disorder. A number of reports claimed familial occurrence and supported single gene inheritance for hypomelanosis of Ito, but none has been proved. Miscellaneous chromosomal mosaicisms have been demonstrated in some but not all affected individuals. Thus, it has been suggested that hypomelanosis of Ito is not a single condition but rather a nonspecific manifestation (ie, a phenotype) of chromosomal mosaicism and that this term should now be dropped. In this article, we review these developments focusing on the neurologic and genetic aspects of hypomelanosis of Ito. Our personal experience with 41 hypomelanosis of Ito patients and literature review led us to conclude that (1) the term hypomelanosis of Ito has been often misapplied to individuals with nonspecific "patchy depigmentation of the skin" who had several conditions of different etiologies; (2) the white matter involvement seen at neuroimaging in most of our hypomelanosis of Ito patients was similar to that reported in well-defined neurocutaneous disorders, including Sjogren-Larsson syndrome and incontinentia pigmenti; (3) whatever figures we take for associated central nervous system abnormalities in hypomelanosis of Ito, these represent the most frequent extracutaneous findings and, therefore, the use of the term neurocutaneous disorder for hypomelanosis of Ito might well be appropriate. PMID- 11063077 TI - Prediction of outcome at school age in neonatal intensive care unit graduates using neonatal neurologic tools. AB - Prediction of outcome for neonatal intensive care unit graduates is clinically useful to counsel families effectively and target those who may benefit from early interventions. Evoked potentials have proven prognostic value of neurologic outcomes in early childhood; however, their long-term predictive validity remains to be determined. The objective of this prospective study was to determine the long-term predictive value of three neonatal neurologic assessments: brainstem auditory evoked potentials, somatosensory evoked potentials, and the Einstein Neonatal Neurobehavioral Assessment Scale. Seventy-eight high-risk newborns and 28 healthy controls were recruited and were assessed in the newborn period using these tests. At 8 to 9 years of age, 42 subjects and 13 controls were re evaluated for developmental progress using a range of psychologic, sensorimotor, and neurologic measures. Findings indicated that the somatosensory evoked potential was most accurate at predicting outcome at school age, with high specificity (83-100%) across all domains tested and good sensitivity (80-100%) for intellectual performance and sensorimotor abilities. The brainstem auditory evoked potential was limited by false-negatives, whereas the neonatal neurobehavioral assessment yielded many false-positives. This study provides new evidence that associations between neonatal somatosensory evoked potentials and developmental sequelae continue to be significant at school age. PMID- 11063078 TI - Usefulness of diagnostic criteria of tuberous sclerosis complex in pediatric patients. AB - The Tuberous Sclerosis Complex 1998 Consensus Conference clinical criteria represent an important advance in the diagnosis of tuberous sclerosis complex. Since many findings regarded as highly specific for tuberous sclerosis complex are not apparent until late childhood or adulthood, refinements by age may prove of value. We have stratified 106 children into five age groups (0 to 2 years of age, above 2 to 5 years, above 5 to 9 years, above 9 to 14 years, and above 14 to 18 years). Physicians should be alerted as to the frequency of the criteria in different stages of children. PMID- 11063079 TI - Psychological functioning in children and adolescents with Sturge-Weber syndrome. AB - Previous studies of individuals with Sturge-Weber syndrome have focused on the medical aspects of this syndrome, but little has been known about the affective and behavioral correlates. We collected psychological and medical data from parents and teachers for 79 children and adolescents with Sturge-Weber syndrome and a group of their siblings. We also obtained the results of intellectual assessment for a subset of the Sturge-Weber syndrome group. The young people with Sturge-Weber syndrome exhibited more problems than the group of siblings across a number of behavioral domains: intellectual/academic, social skills, mood, and compliance. Those children most at risk for psychological problems were those with lower levels of intellectual functioning, those with seizure disorders, and those with more frequent seizures. Larger port-wine stains were also associated with an increase in mood and social problems but only for older children. Increased age was not associated with lower levels of intellectual or academic functioning, but mood and social problems were more common in older children. PMID- 11063080 TI - Postinfectious encephalomyelitis: etiologic and diagnostic trends. AB - Fifty cases of postinfectious encephalomyelitis admitted to our Pediatric Department during the period 1980 to 1997 were consecutively collected and reviewed. There were 28 males and 22 females. The age of onset ranged from 9 months to 14 years. The antecedent infections included measles (6 cases), rubella (5 cases), mumps (4 cases), chicken pox (4 cases), Epstein-Barr virus infection (11 cases), mycoplasma infection (6 cases), and unknown etiology (14 cases). The cessation of measles, rubella, and mumps as causes for encephalomyelitis in our patients corresponds with the introduction of a measles-mumps-rubella nationwide vaccination program in Taiwan commencing in 1992. The main clinical symptoms were fever, headache, and/or vomiting, seizure, and motor weakness. The presenting signs included altered consciousness, meningeal signs, cranial nerve palsy, brainstem signs, involuntary movement, and cerebellar signs. Computed tomography scans were abnormal for 14 (56%) of 25 patients studied, whereas magnetic resonance imaging (MRI) disclosed lesions in 14 (82%) of 17 patients, with abnormal signals in various parts of the cerebral hemisphere, as well as in the basal ganglion, diencephalon, midbrain, brain stem, and cerebellum. Of the three patients with negative MRI findings, an abnormal finding on somatosensory evoked potential was noted for one patient, and a focal decrease in tracer uptake on single photon emission computed tomography (SPECT) was found for the other two patients. This study demonstrates that the causative agents of postinfectious encephalomyelitis in Taiwan have changed from those of traditional exanthematous diseases to nonspecific respiratory infections and suggests that this may also be the case in other parts of the world. MRI remains the imaging method of choice, whereas other neurofunctional studies such as evoked potentials and SPECT are complementary for the diagnosis. PMID- 11063081 TI - Add-on lamotrigine treatment in children and young adults with severe partial epilepsy: an open, prospective, long-term study. AB - We evaluated the efficacy and safety of lamotrigine in 41 children and young adults (age range, 3-25 years; mean, 12 years) with drug-resistant, partial epilepsies, based on a prospective, add-on study. Patients had severe symptomatic/cryptogenic partial epilepsies (mean seizure frequency = 3.6/day), resistant to one to four major antiepileptic drugs. Mean seizure frequency significantly decreased (P < .001) throughout the period of treatment. A good response (>50% seizure reduction) was observed in 15 patients of whom 6 were seizure-free (follow-up: 12-48 months). Higher responder rate was found among cryptogenic epilepsies and epilepsies symptomatic of cerebral malformation, whereas patients with posthypoxic-ischemic perinatal damage were poor responders. Lamotrigine discontinuation was mainly due to lack of efficacy (46% of patients), whereas only 2 patients developed a transient skin rash and did not drop out. Lamotrigine represents a valuable treatment for severe partial epilepsies of childhood that have proved resistant to previous antiepileptic drugs. PMID- 11063082 TI - Molecular genetic classification of central nervous system malformations. AB - Traditional schemes of classifying nervous system malformations are based on descriptive morphogenesis of anatomic processes of ontogenesis, such as neurulation, neuroblast migration, and axonal pathfinding. This proposal is a first attempt to incorporate the recent molecular genetic data that explain programming of development etiologically. A scheme based purely on genetic mutations would not be practical, in part because only in a few dysgeneses are the specific defects known, but also because several genes might be involved sequentially and many genes inhibit or augment the expression of others. The same genes serve different functions at different stages and are involved in multiple organ systems. Some complex malformations, such as holoprosencephaly, result from several unrelated defective genes. Finally, a pure genetic classification would be too inflexible to incorporate some anatomic criteria. The basis for the proposed scheme is, therefore, disturbances in patterns of genetic expression; polarity gradients of the axes of the neural tube (eg, upregulation or downregulation of genetic influences); segmentation (eg, deletions of specific neuromeres, ectopic expression); mutations that cause change in cell lineage (eg, dysplastic gangliocytoma of cerebellum, myofiber differentiation within brain); and specific genes or molecules that mediate neuroblast migration in its early (eg, filamin-1), middle (eg, LIS1, double-cortin), or late course (eg, reelin, L1 CAM). The proposed scheme undoubtedly will undergo many future revisions, but it provides a starting point using currently available data. PMID- 11063083 TI - Pediatric neurology in Israel. PMID- 11063084 TI - Siblings with cystic leukoencephalopathy and megalencephaly. AB - Cystic leukoencephalopathy with megalencephaly is a newly described entity with mild clinical involvement. Patients suffer from developmental problems and seizures in childhood. Progression is gradual into adulthood. Typical magnetic resonance imaging findings include subcortical cysts and diffuse leukoencephalopathy. The etiology is unknown with possibly autosomal-recessive inheritance. We present two pairs of siblings with this disease and emphasize the characteristic and variable patterns even within the same family. PMID- 11063085 TI - Vertebral artery dissection and migraine headaches in children. AB - Strokes of the posterior circulation are uncommon in childhood. In vertebrobasilar insults, vertebral artery dissection remains a rare diagnosis. We report the case of an 8-year-old boy with a history of migraine headaches who presented with acute cerebellar signs and agitation following multiple infarctions of bilateral cerebellar hemispheres. Vertebral angiography demonstrated dissection of the left vertebral artery with occlusion of the basilar artery just distal to its origin. Risk factors for vertebral artery dissection are reviewed, with emphasis on association with migraine headaches. A review of imaging studies for the diagnosis of dissection is also presented. This case demonstrates the importance of considering arterial wall dissection in pediatric patients with a history of atypical migraines associated with new neurologic findings. PMID- 11063086 TI - Early-onset Moyamoya syndrome in a patient with Down syndrome: case report and review of the literature. AB - Moyamoya disease is a chronic occlusive cerebrovascular disorder. It can occur as a primary disease or as a syndrome associated with a variety of conditions. Usually it takes 1 to 2 years to develop a classic moyamoya pattern. We report a 20-month-old girl with Down syndrome and moyamoya syndrome who presented with seizure and hemiparesis. To our knowledge, this is the youngest case reported with moyamoya syndrome and Down syndrome. The prognosis and current treatment of moyamoya syndrome and its relation to Down syndrome are reviewed. There is some reason to speculate that the abnormalities associated with Down syndrome might create a vulnerability for the development of moyamoya syndrome. PMID- 11063087 TI - Asperger's syndrome, X-linked mental retardation (MRX23), and chronic vocal tic disorder. AB - Pervasive developmental disorders are severe disorders of development with no consistent neurobiologic etiology and most often an idiopathic etiology. We report a 12-year-old male who met criteria for a pervasive developmental disorder (Asperger's syndrome) and a chronic tic disorder. The child also has an X-linked cognitive impairment (MRX23). The presence of tic symptomatology, pervasive developmental disorder, and fragile X syndrome has previously been reported. Since no singular etiology for Asperger's syndrome has been found, the possibility of other cases of Asperger's syndrome occurring with concurrent abnormalities on the X chromosome should be considered by clinicians, especially if tic symptomatology is present. PMID- 11063088 TI - Liposteroid therapy for refractory seizures in children. AB - Liposteroid is dexamethasone palmitate incorporated into liposomes and was developed as an anti-inflammatory drug for targeting therapy mainly for rheumatoid arthritis. Recently, it was reported that liposteroid might be effective for the treatment of West syndrome, with fewer side effects than those of corticotropin therapy. We describe three patients, a 2-month-old boy with early infantile epileptic encephalopathy, a 4-month-old girl with symptomatic West syndrome, and a 2-year-old girl with symptomatic localization-related epilepsy, whose refractory seizures were treated with liposteroid according to the original method reported by Yamamoto and colleagues in 1998. Uncontrollable seizures ceased completely in two patients and the seizure frequency decreased markedly in the other patient. Electroencephalograms revealed marked improvement in all patients. They showed no relapse of the seizures, and all showed no adverse effects except for mild brain shrinkage in one patient. Our experience with these three patients suggests that liposteroid therapy might be a new option for the treatment of refractory seizures in children, as well as for West syndrome. PMID- 11063089 TI - Intravascular circulation and distribution of human 51Cr-DBBF stroma-free hemoglobin, 51Cr-plasma, 51Cr-saline, 59FE-plasma, and 125I-albumin in the mouse. AB - Male B6C3HF1 mice were infused with human 51Cr-labeled DBBF (bis 3,5 dibromosalicyl fumarate) crosslinked stroma-free hemoglobin (SFH). In the first hour following SFH infusion, 11.2% of the infused radioactivity was found in the skin, 11.4% in muscle, 9.1% in the skeleton, and 5% in the liver. Twenty-four hours after infusion, 15.4% of the radioactivity was found in the skin, 10.3%, in the muscle, 16.6% in the skeleton, and 6.7% in the liver. The circulation and distribution of 51Cr-labeled DBBF-SFH were compared with levels of 51Cr labeled plasma, 51Cr in saline, 59Fe labeled plasma, and 125I albumin. The radioactivity in the blood was similar for 51Cr-DBBF-SFH, 51Cr-plasma, and 59Fe-plasma. During the 24-hour post-infusion period, extravascular distribution of the 51Cr-saline, 51Cr-plasma, and 125I albumin within the organs was similar to that of 51Cr-DBBF SFH, with the highest levels being in skin, muscle, skeleton and liver, and no increase in the levels in the lung or spleen. The distribution of 59Fe compared to that of 51Cr-DBBF, 51Cr-plasma, 51Cr-saline, and 125I albumin can be explained by the fact that 59Fe is utilized in the production of new red blood cells. PMID- 11063090 TI - Redox concerns in the use of acellular hemoglobin-based therapeutic oxygen carriers: the role of plasma components. AB - Within the past decade, most research efforts in the red blood cell substitute area have revolved about the development of acellular hemoglobin-based oxygen carriers (HBOC) as clinical replacements and/or augmentation of human blood's carrying and delivery function. A major requirement for all HBOC is the maintenance of the heme-Fe+2 in this reduced state for normal physiological behavior. Oxidation of hemoglobin results in the formation of methemoglobin (heme Fe+3). MetHb is unable to bind oxygen thus effectively lowering the carrying capacity of the Hb-based substitute. In addition, met Hb gives rise to free radicals that have the potential to cause endothelial and surrounding tissue damage. Results of this study suggest that the normal endogenous reducing agents of human plasma have the capacity to provide redox protection and stability to specific acellular-types of HBOC. The effectiveness of these reducing agents may be related to the formal reduction potential of the HBOC being considered. The choice of buffer for HBOC storage is critical and specific to the HBOC product. PMID- 11063091 TI - Studies on the resuscitative efficacy of polymerized bovine hemoglobin. AB - The paper described modified hemoglobin (Hb) with glycolaldehyde for the efficacy on resuscitation of severe hemorrhagic shock. Our objective was to compare the effect on resuscitation of severe hemorrhagic shock with different experimental groups. Results showed early resuscitation with modified bovine Hb is superior to lactated Ringer's solution group in improving hemodynamic and acidosis, but the effect of polymerized Hb with glycolaldehyde were similar to fresh whole blood in this rat model. PMID- 11063092 TI - Immobilization of hormones for drug targeting. AB - Biological active compounds such as insulin, heparin, progesterone and labeled-LH were entrapped in glutaraldehyde cross-linked bovine serum albumin (BSA) and human serum albumin (HAS) microspheres. Studies were carried out for their binding capacity and biodegradability using new proteolytic enzymes. Effects of proteolytic enzymes such as trypsin, chymotrypsin, papain and pronase-E on microspheres were studied in order to understand the biodegradability of the cross-linked proteins. It has been observed that labeled-LG was entrapped 60% in BSA and HAS microspheres. Labelled-LH-BSA, Labelled-LH-HAS and insulin microspheres were injected into mice and rabbits. It was observed that these cross-linked microspheres were biodegradable and the process appeared to be slow one, useful for sustained release of hormones. It was also observed that these albumin microspheres exhibit fluorescence at 495 nm. PMID- 11063093 TI - Iodinated natural rubber latex: preparation, characterisation & antibacterial activity assessment. AB - Natural Rubber (NR) in both its latex and dried form was treated with iodine to make it antibacterial in nature. The NR latex was dried and washed, dissolved in toluene, iodinated and cast into films. In a different approach, NR in its latex form (emulsion) was blended with aqueous solution of povidone-iodine complex (PVP I) and films were cast. Phase separation of PVP-I was observed in the films prepared using NR latex; whereas the film prepared using NR solution in toluene and molecular iodine were homogeneous, transparent light brown. Solubility assessment of the films revealed that unlike pure NR, the iodinated NR failed to go into solution completely. The films obtained from both the approaches were evaluated for release of iodide ions. On immersion in water, PVP-I blended latex did not retain iodine while sustained release of iodide ion was observed in case of dried NR dissolved in toluene and treated with iodine. The FT-IR spectra of the iodinated films revealed that iodine attaches covalently to the double bonds in the isoprene units of NR. The films were also evaluated for their antibacterial properties and it was observed that the films prepared from both the approaches, acquired antimicrobial properties. PMID- 11063094 TI - On bioartificial liver assist system: theoretical exploration and strategies for further development. AB - The major difficulty in establishing a clinical effective bioartificial liver assist device for treatment of fulminate hepatic failure is limitation of our knowledge and technologies about fresh cell behaviors in culture and a lack of knowledge about the etiology and pathogenesis of hepatic coma. Increasing data from clinical and laboratory investigation have accrued indicating that toxins from necrotic liver tissue, mainly as oxygen reactive substances, have a role in the pathogenesis of hepatic encephalopathy and even multiple system organs failure. This paper presents the data available and suggests a new pathway for artificial and bioartificial liver assist system. PMID- 11063095 TI - Cerebrovascular surgery: past, present, and future. PMID- 11063096 TI - Molecular biology of degenerative disc disease. AB - The intervertebral disc is a complex anatomic and biochemical structure. It is composed primarily of fibrocytes and chondrocytes that are anatomically segregated in an elaborate avascular macromolecular matrix of collagen and proteoglycans. Degenerative processes associated with aging and trauma result in morphological and molecular changes to the disc. Morphological changes are observed as dehydration, fissuring, and tearing of the nucleus, annulus and endplates. On the molecular level, degenerative changes include decreased diffusion, decreased cell viability, decreased proteoglycan synthesis, and alteration in collagen distribution. The role of inflammatory mediators in these processes, and the potential use of growth factors to delay or reverse the degenerative cascade, is poorly understood. However, these areas are under active investigation, the results of which may soon contribute significantly to our understanding of degenerative disc disease. PMID- 11063097 TI - Electrical bone graft stimulation for spinal fusion: a review. AB - Although electrical stimulation to aid bone fusion is well established in the treatment of long-bone fractures, its use as an aid in spinal fusion is not as well documented. This article presents the history and scientific basis of electrical stimulation to aid bone fusion and extensively reviews the clinical literature. It is intended to provide an objective review of the indications and limitations of electrical stimulation to enhance spinal fusion and to serve as a reference source for further study. PMID- 11063098 TI - Olfactory ensheathing cells: bridging the gap in spinal cord injury. AB - Spinal cord injury (SCI) continues to be an insidious and challenging problem for scientists and clinicians. Recent neuroscientific advances have changed the pessimistic notion that axons are not capable of significant extension after transection. The challenges of recovering from SCI have been broadly divided into four areas: 1) cell survival; 2) axon regeneration (growth); 3) correct targeting by growing axons; and 4) establishment of correct and functional synaptic appositions. After acute SCI, there seems to be a therapeutic window of opportunity within which the devastating consequences of the secondary injury can be ameliorated. This is supported by several observations in which apoptotic glial cells have been identified up to 1 week after acute SCI. Moreover, autopsy studies have identified anatomically preserved but unmyelinated axons that could potentially subserve normal physiological properties. These observations suggest that therapeutic strategies after SCI can be directed into two broad modalities: 1) prevention or amelioration of the secondary injury, and 2) restorative or regenerative interventions. Intraspinal transplants have been used after SCI as a means for restoring the severed neuraxis. Fetal cell transplants and, more recently, progenitor cells have been used to restore intraspinal circuitry or to serve as relay for damaged axons. In an attempt to remyelinate anatomically preserved but physiologically disrupted axons, newer therapeutic interventions have incorporated the transplantation of myelinating cells, such as Schwann cells, oligodendrocytes, and olfactory ensheathing cells. Of these cells, the olfactory ensheathing cells have become a more favorable candidate for extensive remyelination and axonal regeneration. Olfactory ensheathing cells are found along the full length of the olfactory nerve, from the basal lamina of the epithelium to the olfactory bulb, crossing the peripheral nervous system-central nervous system junction. In vitro, these cells promote robust axonal growth, in part through cell adhesion molecules and possibly by secretion of neurotrophic growth factors that support axonal elongation and extension. In animal models of SCI, transplantation of ensheathing cells supports axonal remyelination and extensive migration throughout the length of the spinal cord. Although the specific properties of these cells that govern enhanced axon regeneration remain to be elucidated, it seems certain that they will contribute to the establishment of new horizons in SCI research. PMID- 11063099 TI - Quantification of, visualization of, and compensation for brain shift using intraoperative magnetic resonance imaging. AB - OBJECTIVE: Modern neuronavigation systems lack spatial accuracy during ongoing surgical procedures because of increasing brain deformation, known as brain shift. Intraoperative magnetic resonance imaging was used for quantitative analysis and visualization of this phenomenon. METHODS: For a total of 64 patients, we used a 0.2-T, open-configuration, magnetic resonance imaging scanner, located in an operating theater, for pre- and intraoperative imaging. The three-dimensional imaging data were aligned using rigid registration methods. The maximal displacements of the brain surface, deep tumor margin, and midline structures were measured. Brain shift was observed in two-dimensional image planes using split-screen or overlay techniques, and three-dimensional, color coded, deformable surface-based data were computed. In selected cases, intraoperative images were transferred to the neuronavigation system to compensate for the effects of brain shift. RESULTS: The results demonstrated that there was great variability in brain shift, ranging up to 24 mm for cortical displacement and exceeding 3 mm for the deep tumor margin in 66% of all cases. Brain shift was influenced by tissue characteristics, intraoperative patient positioning, opening of the ventricular system, craniotomy size, and resected volume. Intraoperative neuronavigation updating (n = 14) compensated for brain shift, resulting in reliable navigation with high accuracy. CONCLUSION: Without brain shift compensation, neuronavigation systems cannot be trusted at critical steps of the surgical procedure, e.g., identification of the deep tumor margin. Intraoperative imaging allows not only evaluation of and compensation for brain shift but also assessment of the quality of mathematical models that attempt to describe and compensate for brain shift. PMID- 11063100 TI - Near-real-time guidance using intraoperative magnetic resonance imaging for radical evacuation of hypertensive hematomas in the basal ganglia. AB - OBJECTIVE: To report our preliminary clinical experience in treating patients with hypertensive hemorrhage in the basal ganglia using a minimally invasive approach facilitated by intraoperative real-time imaging of an open magnetic resonance imaging (MRI) system and a newly designed cutting suction device. METHODS: We developed an artifact-free device for use during intraoperative MRI consisting of a guiding base that locks into a burr hole, a side-cutting composite-based cannula connected to a standard aspirator, and a handpiece that allows aspiration strength to be regulated by the surgeon. Thirteen patients with hypertensive bleeding in the basal ganglia were included in the study. Outcome was evaluated by mortality, Glasgow Outcome Scale score, activities of daily living score, and Rankin score at 2 weeks and at a median of 4.2 months after the hemorrhage. RESULTS: In this group of 13 patients, complete evacuation was achieved in 8 patients (62%) and subtotal evacuation of 75 to 90% of the initial volume in 4 patients (31%); the evacuation was partial in 1 patient (8%). Vascular malformations were preoperatively excluded angiographically. There was no rebleeding during surgery or postoperatively, as demonstrated by immediate postoperative MRI and computed tomography on the 1st postoperative day. Hematomas were evacuated on median Day 4 after the hemorrhage, varying between Day 1 and Day 8; evacuation was performed on Day 21 after the hemorrhage in one patient. Twelve of the 13 patients survived during a median follow-up time of 4.2 months. Neurological function improved in 11 of the 12 patients eligible for assessment. One patient with an additional head injury died 15 days after surgery from pulmonary embolism. CONCLUSION: This study shows an excellent outcome with regard to mortality and a positive trend regarding neurological outcome for the specific group of patients with hypertensive hematomas in the basal ganglia. This minimally invasive approach is feasible in the open intraoperative MRI in combination with the cutting suction device developed in our institution. Online imaging is extremely helpful for planning, guiding, and real-time monitoring of the procedure. PMID- 11063101 TI - Radiosurgery for epilepsy associated with cavernous malformation: retrospective study in 49 patients. AB - OBJECTIVE: Microsurgical resection of a cavernous malformation (CM) with or without associated cortical resection can provide efficient treatment of drug resistant associated epilepsy. To explore the potential alternative role of radiosurgery and to evaluate its safety and efficacy for this indication, we conducted a retrospective multicenter study. METHODS: We retrospectively reviewed the files of patients with long-lasting drug-resistant epilepsy, presumably caused by CM, who were treated by gamma knife (GK) surgery for the control of their epilepsy in five centers (Marseilles, Komaki City, Prague, Graz, and Sheffield). A satisfactory follow-up was available for 49 patients (mean follow up period, 23.66 +/- 13 mo). The mean duration of epilepsy before the GK procedure was 7.5 (+/-9.3) years. The mean frequency of seizures was 6.9/month (+/-14). The mean marginal radiation dose was 19.17 Gy +/- 4.4 (range, 11.25-36). Among the 49 patients, 17 (35%) had a CM located in or involving a highly functional area. RESULTS: At the last follow-up examination, 26 patients (53%) were seizure-free (Engel's Class I), including 24 in Class IA (49%) and 2 patients with occasional auras (Class IB, 4%). A highly significant decrease in the number of seizures was achieved in 10 patients (Class IIB, 20%). The remaining 13 patients (26%) showed little or no improvement. The mediotemporal site was associated with a higher risk of failure. One patient bled during the observation period, and another experienced radiation-induced edema with transient aphasia. Postradiosurgery excision was performed in five patients, and a second radiosurgical treatment was carried out in one patient. CONCLUSION: This series is the first to specifically evaluate the capability of GK surgery to safely and efficiently treat epilepsy associated with CM. Seizure control can be reached when a good electroclinical correlation exists between CM location and epileptogenic zone. Although we do not recommend GK surgery for prevention of bleeding for a CM that has not bled previously, our findings suggest that GK surgery can be proposed for the treatment of epilepsy when the CM is located in a highly functional area. PMID- 11063102 TI - Neurological grades of patients with poor-grade subarachnoid hemorrhage improve after short-term pretreatment. AB - OBJECTIVE: Short-term pretreatment of patients with subarachnoid hemorrhage, but without hematomas causing mass effect, who presented in poor neurological condition at admission was evaluated as a protocol for the selection of candidates for radical surgery. METHODS: One hundred-three patients were pretreated for 12 hours with control of blood pressure and intracranial pressure, using diuretic agents and/or ventricular drainage. RESULTS: Neurological improvement was observed for 32 of 47 patients in Grade IV at admission and 23 of 56 patients in Grade V (P < 0.01). Hydrocephalus requiring drainage was more common (P < 0.05) and the interval between onset and admission was shorter (P < 0.01) for the improved group. Clipping surgery was performed for all patients in Grade III or better and for patients in Grade IV who were less than 75 years of age and without systemic complications, i.e., 38 of 47 patients in Grade IV and 16 of 56 patients in Grade V at admission. Good outcomes (defined as moderately disabled or better on the Glasgow Outcome Scale) were achieved by 34 of 38 patients in Grade IV and 10 of 16 patients in Grade V (P < 0.01). The proportion of patients in Grade IV after pretreatment was lower for Grade IV (2 of 38 patients) than for Grade V (9 of 16 patients) (P < 0.00001). However, none of the 49 patients who underwent nonsurgical treatment achieved good outcomes. CONCLUSION: Our protocol may be beneficial for the selection of candidates for radical surgery among patients with subarachnoid hemorrhage but without hematomas who are in poor neurological condition at admission and for the improvement of postoperative outcomes. PMID- 11063103 TI - Multiple interstitial substances measured by microdialysis in patients with subarachnoid hemorrhage. AB - OBJECTIVE: Intracerebral microdialysis is a tool to monitor metabolic disturbances in the brains of patients with severe head injuries or subarachnoid hemorrhage (SAH). In the search for putative indicators of primary and secondary brain damage, we measured multiple metabolites in the dialysates of patients with SAH, to elucidate their significance for the outcomes of the patients as well as their temporal profiles of liberation after the insult. METHODS: Microdialysis probes were placed, with a ventriculostomy catheter for drainage of cerebrospinal fluid, into a frontal lobe of 10 patients with aneurysmal SAH, for 4.6 +/- 0.5 days. Amino acids, metabolites of glycolysis, purines, catecholamines, and nitric oxide oxidation byproducts were measured by high-performance liquid chromatography. Spearman's correlation coefficient and Student's t test were used to compare the levels of the metabolites with the outcomes of the patients, as assessed using the Glasgow Outcome Scale, 3 months after the ictus. RESULTS: For patients with unfavorable outcomes (Glasgow Outcome Scale scores of 1-3), which were primarily associated with the development of large infarctions, dialysate levels of excitatory amino acids increased up to 30-fold, those of lactate up to 10-fold, and those of nitrite up to 5-fold, compared with normal levels observed for patients with favorable outcomes (Glasgow Outcome Scale scores of 4 or 5). When average peak concentrations in the dialysates of patients with favorable and unfavorable outcomes were compared, significantly higher levels of excitatory amino acids, taurine, lactate, and nitrite, but not of purines and catecholamines, were observed for those with poor outcomes (P < 0.05). With respect to the temporal profiles of the average metabolite concentrations, the significantly increased levels of amino acids observed for patients with poor outcomes followed a biphasic course, with maximal concentrations on the first and second days or the seventh day after the insult (P < 0.01). CONCLUSION: These data confirm the usefulness of excitatory amino acids and lactate as major parameters for neurochemical monitoring for patients threatened by acute cerebral disorders. Other substances, such as taurine and nitrite, were also demonstrated to be potentially predictive. Release of these substances into the extracellular fluid of the brain might be particularly relevant for the development of secondary brain damage after SAH, e.g., infarction or brain swelling. PMID- 11063104 TI - Transcranial brainstem stab injuries: a retrospective analysis of 17 patients. AB - OBJECTIVE: Transcranial stab injuries remain a frequent cause of emergent neurosurgical admissions to neurosurgical units in South Africa. Brainstem stabs are an uncommon, yet often fatal, form of brain injury. METHODS: A retrospective audit of 597 patients with transcranial stab injuries admitted to our unit over a 12-year period (January 1987 to December 1998) identified 17 patients (2.85%) with brainstem stab injuries. The computed tomographic scans of all patients were analyzed, and a detailed autopsy examination of the skull and its contents was performed in all patients who died. Stepwise linear regression analysis was used to formulate a predictive model of outcome for the entire series of 597 patients. RESULTS: The majority of the patients were males (16 patients), and the study group had a mean age of 28.65 +/- 9.59 years and a mean Glasgow Coma Scale score of 8.59 +/- 2.76. Knives (82%) were the most common instruments of penetration. Cerebral angiography identified 3 patients with vascular abnormalities, and autopsy revealed an additional 4 patients with vascular injury. Emergency ventriculostomy was performed in 10 patients for obstructive hydrocephalus. Four of the 17 patients survived (76.5% mortality). Factors significantly predictive of outcome in patients with transcranial stab injuries were the Glasgow Coma Scale score (F = 43.7), the occurrence of intraventricular hemorrhage (F = 22.8), the type of associated lesion (intracranial bleed, vascular abnormality, or brain abscess) (F = 5.9), and the number of operations (F = 3.2). CONCLUSION: The Glasgow Coma Scale score is the most significant predictor of outcome in low velocity transcranial stab injuries. Brainstem stab injuries have a great propensity for vascular damage. Survivors are incapacitated by severe, fixed neurological deficits. PMID- 11063105 TI - The efficacy and cost of prophylactic and perioprocedural antibiotics in patients with external ventricular drains. AB - OBJECTIVE: Prophylactic antibiotics are routinely administered to patients with external ventricular drains (EVDs); however, no conclusive evidence supports this practice. This study compared the efficacy and cost of prophylactic and periprocedural antibiotics in patients with EVDs. METHODS: We reviewed the charts of 308 patients who had an EVD in place for 3 or more days between January 1996 and June 1997. Patients with EVDs placed for shunt infections or meningitis were excluded. A standard protocol was used to insert and monitor EVDs. Catheters were left in place as long as clinically indicated and changed only if they malfunctioned. Cerebrospinal fluid cultures were obtained twice weekly. Prophylactic antibiotics were used at the discretion of the attending neurosurgeon. Patients were divided into two groups: Group A comprised 209 patients who received prophylactic antibiotics for the duration of the EVD (intravenously administered cefuroxime, 1.5 g every 8 h); Group B comprised 99 patients who received only periprocedural antibiotics (intravenously administered cefuroxime, 1.5 g every 8 h, three or less doses). RESULTS: Although there were significantly more males in Group A than in Group B, the two groups were otherwise well matched, with no significant differences in age, indications, or duration of EVD placement. The overall rate of ventriculitis was 3.9%. The infection rates for Group A (3.8%) and Group B (4.0%) were almost identical. CONCLUSION: Prophylactic antibiotics did not significantly reduce the rate of ventriculitis in patients with EVDs, and they may select for resistant organisms. Discontinuing the use of prophylactic antibiotics for EVDs at the authors' institution would save approximately $80,000 per year in direct drug costs. PMID- 11063106 TI - Parameters for contralateral approach to ophthalmic segment aneurysms of the internal carotid artery. AB - OBJECTIVE: This study was undertaken to define more accurately the feasibility and indications of the contralateral pterional approach to ophthalmic segment aneurysms of the internal carotid artery (ICA). METHODS: Between 1995 and 1999, 46 patients with ophthalmic segment aneurysms of the ICA were surgically treated in our institution. Eleven of the 46 aneurysms were operated using the contralateral pterional approach. All aneurysms were successfully clipped without complications; three patients required bone resection around the aneurysm neck. We studied the 11 patients who were treated with the contralateral approach by defining six parameters to assess the feasibility of the approach and to predict the necessity for bone resection: 1) Parameter A, the distance between the anterior aspect of the optic chiasm and the limbus sphenoidale; 2) Parameter B, the distance between the bilateral optic nerves at the entrance to the optic canal; 3) Parameter C, the interrelation of the optic nerve and the ICA, expressed as a/b in which a is the length from the midline to the optic nerve and b is the length from the midline to the ICA; 4) Parameter D, the size of the aneurysm neck; 5) Parameter E, the direction of the aneurysm from the ICA wall on the anteroposterior angiogram; and 6) Parameter F, the distance from the medial side of the estimated distal dural ring to the proximal aneurysm neck on the lateral angiogram. RESULTS: Parameters A to F were 8.8 mm (range, 5.4-11.1 mm), 14.5 mm (range, 10.4-22.2 mm), 0.9 mm (range, 0.6-1.3 mm), and 3.0 mm (range, 2.3 4.7 mm), 5 to 160 degrees, and 1.3 mm (range, 0.3-2.4 mm), respectively. All patients had excellent operative outcomes without visual dysfunction. Three patients required drilling of the bone around the optic canal on the craniotomy side; bone drilling was not required when Parameter E was between 30 and 160 degrees and Parameter F was more than 1 mm. CONCLUSION: Parameters A to D are important for assessing the feasibility of the contralateral approach to ICA ophthalmic segment aneurysms, and Parameters E and F are most useful for calculating the difficulty of this approach. PMID- 11063107 TI - Frameless stereotactic neurosurgery using intraoperative magnetic resonance imaging: stereotactic brain biopsy. AB - OBJECTIVE: To assess the application accuracy of intraoperative magnetic resonance imaging for frameless stereotactic surgery, and to evaluate the performance of intraoperative magnetic resonance imaging for the brain biopsy, a standard stereotactic procedure. METHODS: A series of spatial coordinate and phantom experiments were performed to analyze the application accuracy of the system. A prospective analysis of 68 consecutive patients undergoing stereotactic brain biopsy was then performed. RESULTS: The spatial coordinate experiments revealed a mean overall error in acquisition of 0.2 mm. The phantom experiments demonstrated a 1:1 correlation between the magnetic resonance image of a stereotactically guided probe and its relationship to a target and the actual relationship of the probe and target. Sixty-eight brain biopsies were successfully performed in all intracranial compartments except the sella. The radiographic abnormality was localized successfully in all patients (100%). Sixty six (97.1%) of the biopsies yielded diagnostic tissue. Two biopsies (2.9%) were complicated by intraparenchymal hemorrhage. One expanding temporal lobe hemorrhage was evacuated by immediate craniotomy in the magnet with no postoperative sequelae. A deep hemorrhage from a lymphoma was managed conservatively with interval resolution of symptoms. There were no infections. There was no perioperative mortality. CONCLUSION: Intraoperative magnetic resonance imaging allows excellent target localization, provides true real-time imaging to account for anatomic changes during surgery, and permits intraoperative confirmation that the biopsy needle has reached the targeted lesion. Immediate postoperative imaging in the operating room allows assessment of adverse events and the potential for immediate management of hemorrhagic complications. PMID- 11063108 TI - Endovascular treatment of distally located giant aneurysms. AB - OBJECTIVE: Because giant aneurysms (GAs) can be technically difficult to clip, the endovascular approach is becoming increasingly popular. Endovascular treatment of distally located GAs, which often requires parent vessel occlusion, is particularly challenging because limited pathways are available for collateral flow. We aimed to determine the outcomes of endovascular attempts to treat GAs downstream from the circle of Willis. METHODS: Between 1991 and 1998, 27 patients with 27 distally located very large aneurysms or GAs were evaluated for possible endovascular treatment. Ten underwent selective embolization and 9 were treated with primary parent vessel occlusion, with or without distal bypass. Eight patients could not be treated endovascularly. RESULTS: Selective embolization resulted in only one cure. Two patients died as a result of subarachnoid hemorrhage during the follow-up period. One coil-treated patient, who underwent subsequent spontaneous parent vessel occlusion, and all nine patients treated primarily with parent vessel occlusion were considered cured after their treatments. Only two patients treated with parent vessel occlusion experienced periprocedural ischemia, which did not result in a major deficit in either case. Of the eight patients who could not be treated endovascularly, one succumbed to surgery, four died while being treated conservatively, and three were lost to follow-up monitoring. CONCLUSION: Selective aneurysm embolization is usually not curative in these situations. For selected patients, however, endovascular parent vessel occlusion is usually safe and effective in preventing the progression of symptoms and bleeding. PMID- 11063109 TI - Soleus neurotomy for treatment of the spastic equinus foot. Groupe d'Evaluation et de Traitement de la Spasticite et de la Dystonie. AB - OBJECTIVE: This prospective, nonrandomized, noncontrolled study was performed to evaluate the results of a new type of neurotomy, namely the soleus neurotomy, for treatment of the spastic equinus foot. METHODS: Between May 1996 and March 1998, 46 patients were treated for a spastic equinus foot. Clinical status, spasticity (Ashworth Scale score), and kinematic parameters of the gait were determined before and after surgery. The neurotomy was performed on the upper nerve of the soleus in all cases and was associated with other neurotomies (lower nerve of the soleus, 21 patients; gastrocnemius, 9 patients, tibialis posterior, 18 patients; flexor hallucis longus, 16 patients; and flexor digitorum longus, 17 patients). RESULTS: The mean follow-up period was 15 months (range, 8-28 mo). The equinus deformity disappeared clinically in all patients. Before the operation, all patients had an Ashworth Scale score of 2, with an inexhaustible clonus present on knee extension and persisting with knee flexion (Tardieu Scale score, 4), which was abolished in 95% of the patients after surgery. Two patients still had some clonus on knee extension; this did not interfere with their clinical improvement. Knee recurvatum disappeared in eight patients. Analysis of kinematic parameters demonstrated a statistically significant increase in joint motion of the second rocker (P = 0.0026) of the ankle during stance. The duration of the stance or swing phase, length of the walking cycle, and velocity or rate of spontaneous walking were not significantly modified. CONCLUSION: The study demonstrated that soleus neurotomy is effective for the treatment of spastic equinus foot, leading to abolition of spasticity and improvement in the range of ankle motion during the stance phase of gait. PMID- 11063110 TI - Surgical anatomic evaluation of the cervical pedicle and adjacent neural structures. AB - OBJECTIVE: Although several clinical applications of transpedicular screw fixation in the cervical spine have been documented recently, few anatomic studies concerning the cervical pedicle are available. This study was designed to evaluate the anatomy and adjacent neural relationships of the middle and lower cervical pedicle (C3-C7). The main objective is to provide accurate information for transpedicular screw fixation in the cervical region and to minimize complications by providing a three-dimensional orientation. METHODS: Twenty cadavers were used to observe the cervical pedicle and its relationships. After removal of the posterior bony elements, including spinous processes, laminae, lateral masses, and inferior and superior facets, the isthmus of the pedicle was exposed. Pedicle width, pedicle height, interpedicular distance, pedicle-inferior nerve root distance, pedicle-superior nerve root distance, pedicle-dural sac distance, medial pedicle-dural sac distance, mean angle of the pedicle, root exit angle, and nerve root diameter were measured. RESULTS: The results indicate that there was no distance between the pedicle and the superior nerve root and between the pedicle and the dural sac in 16 specimens, whereas there was a slight distance in the lower cervical region in the 4 other specimens. The mean distance between the pedicle and the inferior nerve root for all specimens ranged from 1.0 to 2.5 mm. The mean distance between the medial pedicle and the dural sac increased consistently from 2.4 to 3.1 mm. At C3-C7, the mean pedicle height ranged from 5.2 to 8.5 mm, and the mean pedicle width ranged from 3.7 to 6.5 mm. Interpedicular distance ranged from 21.2 to 23.2 mm. The mean root exit angle ranged from 69 to 104 degrees, with the largest angle at C3 and the smallest at C6. The mean angle of the pedicle ranged from 38 to 48 degrees. The nerve root diameter increased consistently from 2.7 mm at C3 to 3.8 mm at C6 and then decreased to 3.7 mm at the C7 level. Differences in measurements were considered statistically significant at levels ranging from P < 0.05 to P < 0.01. CONCLUSION: This study indicates that improper placement of the pedicle screw medially and superiorly in the middle and lower cervical spine should be avoided and that the anatomic variations between individuals should be established by measurement. PMID- 11063111 TI - Intraoperative electrophysiological monitoring of oculomotor nuclei and their intramedullary tracts during midbrain tumor surgery. AB - OBJECTIVE: During surgery for intrinsic midbrain lesions, we intraoperatively recorded evoked compound muscle action potentials (ECMAPs) from the extraocular muscles and evaluated how this type of intraoperative electrophysiological monitoring could minimize postoperative oculomotor nerve palsy (ptosis and/or diplopia). METHODS: The ECMAPs were recorded through a spring electrode applied to the extraocular muscle (Method 1, seven cases) or a needle electrode inserted into the superior intraorbital space (Method 2, five cases). The surgeon repeated electrical stimulations whenever tissue of unknown origin was encountered intraoperatively, and this information was used to safely guide surgical resection of the tumors. RESULTS: Using these monitoring techniques, the response free areas were resected and the areas from which ECMAP responses were recorded were avoided. For all 12 patients, ECMAPs were successfully recorded from the extraocular muscles. Ten patients did not exhibit any postoperative deterioration of oculomotor nerve function. Two patients exhibited deterioration of oculomotor nerve function immediately after surgery, which resolved within 1 month. Equally robust ECMAPs could be recorded with Method 2, compared with Method 1. CONCLUSION: Intraoperative ECMAP recordings from the extraocular muscles precisely indicated the locations of the oculomotor nuclei and/or intramedullary oculomotor tracts. Although Method 2 is a more indirect method for recording ECMAPs than is Method 1, Method 2 was equally useful in recording ECMAPs, which seemed to be the summed potentials from the superior rectus muscle and the levator palpebrae superioris muscle. These monitoring techniques are valuable in guiding surgeons to avoid causing inadvertent harm to the oculomotor nuclei and tracts during midbrain surgery, particularly when the neuroanatomic features are distorted by the presence of tumor. PMID- 11063112 TI - Occlusive hyperemia: a radiosurgical phenomenon? AB - OBJECTIVE: Causes of neurological deficits after arteriovenous malformation (AVM) radiosurgery, including hemorrhage, radiation injury, and delayed cyst formation, are described. CONCEPT: Occlusive hyperemia has been described as a reason for neurological deterioration after AVM resection. Thrombosis of draining veins or dural sinuses is thought to cause postoperative bleeding or neurological deficits secondary to venous hypertension. In a similar manner, local hemodynamic changes can occur in the brain adjacent to an AVM after radiosurgery if venous outflow is obstructed. Two patients are presented whose cases demonstrate this phenomenon. CONCLUSION: Patients can experience clinical worsening after AVM radiosurgery from premature thrombosis of draining veins. Local hemodynamic changes could explain why imaging changes thought to be radiation related occur more frequently after radiosurgery of AVMs than of tumors. PMID- 11063113 TI - Alpha(v)beta3 and alpha(v)beta5 integrin expression in meningiomas. AB - OBJECTIVE: Integrins are emerging as alternative receptors capable of mediating several biological functions, such as cell-matrix and cell-cell adhesion, cell migration, signal transduction, and angiogenesis. Two alpha(v) integrins, i.e., alpha(v)beta3 and alpha(v)beta5, play critical roles in mediating these activities, particularly in tumors. No data are available on the expression of these integrins in meningiomas. METHODS: Using Western blot and immunohistochemical analyses with LM609 and PG32, two monoclonal antibodies capable of recognizing the functional integrin heterodimer, we evaluated the expression of alpha(v)beta3 and alpha(v)beta5 integrins in a series of 34 meningiomas of different histological subtypes and grades. We studied their expression in tumor cells and vasculature, as well as the expression of their related angiogenic factors (fibroblast growth factor 2 and vascular endothelial growth factor) and the alpha(v)beta3 ligand vitronectin. RESULTS: Alpha(v)beta3 and alpha(v)beta5 integrins were expressed by neoplastic vasculature and cells. Alpha(v)beta3 and alpha(v)beta5 expression was associated and correlated with that of their respective growth factors (fibroblast growth factor 2 and vascular endothelial growth factor) and microvessel counts and densities. Alpha(v)beta3 was more strongly expressed than alpha(v)beta5 in two cases of histologically benign meningiomas with aggressive clinical behavior. Alpha(v)beta3 expression was associated with that of its related ligand vitronectin and was also evident in small vessels of brain tissue closely surrounding meningiomas. CONCLUSION: Our data demonstrate the expression of alpha(v)beta3 and alpha(v)beta5 integrins in meningioma cells and vasculature. Our findings suggest a role for both of these integrins, and particularly alpha(v)beta3, in meningioma angiogenesis. PMID- 11063114 TI - Effects of a mixture of a low concentration of n-butylcyanoacrylate and ethiodol on tissue reactions and the permanence of arterial occlusion after embolization. AB - OBJECTIVE: Cyanoacrylates are the most commonly used liquid embolic agents. For embolization of arteriovenous malformations, a mixture of a low concentration of n-butylcyanoacrylate (NBCA) and Ethiodol (Savage Laboratories, Melville, NY) has been recommended for deeper penetration of the nidus. Dilution of NBCA, however, might result in different degrees of tissue reaction and might influence the permanence of vessel occlusion, with an increased risk of vessel recanalization. We compared tissue reactions induced by different NBCA/Ethiodol mixtures and analyzed the permanence of their embolic effects. METHODS: NBCA was diluted with Ethiodol to prepare the following standard solutions: Mixture A, low concentration (NBCA/Ethiodol ratio of 20:80); Mixture B, high concentration (50:50). The study was designed in two parts, because tissue reactions after embolization are considered to be a combination of foreign body reactions to solidified material and reactions to the injured blood vessel. Foreign body reactions were studied by intramuscularly injecting both glue mixtures into the backs of 18 rats. Specimens were obtained at various times after implantation. Immunohistochemical analysis and esterase staining were used to detect macrophages and neutrophils, respectively. The densities of these inflammatory cells were calculated and statistically compared. To study the degree of vascular wall injury and the permanence of embolic effects, the renal arteries in 48 rabbits were embolized with NBCA Mixture A or B. Six specimens for each group were obtained at various times after embolization. RESULTS: There was no significant difference in foreign body reactions between groups treated with Mixtures A and B, at any time. However, the macrophage density was larger for both groups at 3 months versus 3 days and for the group treated with Mixture B at 3 months versus 2 weeks. There was no difference in the degree of vessel wall injury. None of the embolized vessels demonstrated evidence of recanalization. CONCLUSION: The low concentration of NBCA induced a tissue response similar to that of the high-concentration form. Embolized vessels exhibited no greater incidence of recanalization. Therefore, embolization of arteriovenous malformations with diluted NBCA may be safe. PMID- 11063115 TI - Adenoviral gene transfer of nitric oxide synthase increases cerebral blood flow in rats. AB - OBJECTIVE: Depletion of nitric oxide may play a role in the development of vasospasm after aneurysmal subarachnoid hemorrhage. Replenishment of nitric oxide might be a useful treatment for vasospasm. Using rats, we performed intracisternal injections of replication-defective adenovirus containing the endothelial nitric oxide synthase (eNOS) gene and determined the localization of and effect on cerebral blood flow of transgene expression. METHODS: Rats underwent baseline measurement of cortical cerebral blood flow using laser Doppler flowmetry. Replication-defective adenovirus containing the Escherichia coli LacZ gene (Ad327beta-Gal, n = 2/time point) or the bovine eNOS gene (AdCD8 NOS, n = 4/time point) or physiological saline solution was injected into the cisterna magna. Cerebral blood flow was measured 1, 2, 4, 7, or 14 days later, and the animals were killed. Expression of beta-galactosidase activity from the LacZ gene was examined by histochemical staining and that of eNOS was examined by polymerase chain reaction assays of messenger ribonucleic acid. Brains were histopathologically examined for inflammation. RESULTS: Beta-galactosidase activity was observed throughout the leptomeninges and in some cells in the adventitia of small subarachnoid blood vessels in the Ad327beta-Gal-injected rats. Messenger ribonucleic acid for eNOS was detected in the leptomeninges and brainstem 1 and 2 days after injection of AdCD8-NOS. Rats injected with Ad327beta Gal or physiological saline solution exhibited decreased cerebral blood flow beginning 2 days after virus injection and lasting up to 14 days after injection. Rats injected with AdCD8-NOS developed significant transient increases in cerebral blood flow 2 days after virus injection, followed by slight decreases in blood flow. There was inflammation in the subarachnoid space of all animals; the inflammation was qualitatively worse in animals injected with Ad327beta-Gal, compared with rats injected with AdCD8-NOS or saline solution. CONCLUSION: Intracisternal injection of replication-defective adenovirus containing the eNOS gene can transiently increase cerebral blood flow. PMID- 11063116 TI - The mystery of the missing Viking helmets. AB - Based on archaeological finds and old Norse literature, this study describes the Scandinavian helmet tradition from the Bronze Age to the Viking Age, as well as the Viking culture, with special emphasis on weaponry and head protection. Contrary to what is commonly believed, the study shows that metal helmets must have been used very infrequently by the Vikings. In fact, only one Viking helmet has been retrieved in Scandinavia. Possible reasons for the widespread misconception that the Vikings wore helmets are discussed, and the responsibility for not correcting this misunderstanding is placed with the archaeological profession. PMID- 11063117 TI - Idiopathic hypertrophic cranial pachymeningitis associated with a dural arteriovenous fistula involving the straight sinus: case report. AB - OBJECTIVE AND IMPORTANCE: Reports of idiopathic hypertrophic cranial pachymeningitis have increased as a result of advances in magnetic resonance imaging. This is the first documented case of idiopathic hypertrophic cranial pachymeningitis associated with a dural arteriovenous fistula involving the straight sinus. We discuss possible causes of the association and the treatment options. CLINICAL PRESENTATION: A 64-year-old man presented with a headache and visual disturbance. Gadolinium-enhanced T1-weighted magnetic resonance imaging demonstrated homogeneously stained meninges and prominent enhancement of the tentorium and falx. Angiograms demonstrated a dural arteriovenous fistula of the straight sinus. INTERVENTION: Although surgical excision of the straight sinus and subsequent corticosteroid therapy failed to relieve the patient's visual symptoms, subsequent surgical decompression of the optic nerve resulted in improvement and stabilization. CONCLUSION: Narrowing or occlusion of the tentorial sinuses and narrowing of the straight sinus by extensive dural fibrosis of the tentorium and falx, attributable in turn to idiopathic hypertrophic cranial pachymeningitis, may have resulted in the development of a dural arteriovenous fistula. We propose surgical decompression of the optic nerve as an alternative treatment during the active phase of the disease in patients who exhibit resistance to corticosteroid therapy. PMID- 11063118 TI - Fibrous connective tissue lesion mimicking a vestibular schwannoma: case report. AB - OBJECTIVE AND IMPORTANCE: Cerebellopontine angle fibromas are rare pathological entities that can mimic the presentation of vestibular schwannomas (VSs). Diagnosis of these benign lesions, however, is important, because treatment options may be different. The clinical, radiological, and intraoperative features of these unusual lesions of the cerebellopontine angle are discussed, with review of the relevant literature. CLINICAL PRESENTATION: A 41-year-old man presented with recurrent episodes of diminished hearing on the left side, accompanied by facial ticks and pain on the same side. Magnetic resonance imaging and computed tomographic scans revealed a 1.5-cm, primarily intracanalicular lesion, suggesting a left VS. INTERVENTION: The lesion was partially removed through a retrosigmoid suboccipital craniotomy. Its intraoperative appearance and hard fibrotic consistency differed from the classic features of VSs. The pathological findings indicated nontumoral fibrous connective tissue. The lesion exhibited no features of inflammation or fat and was also negative for S-100 staining. Follow up magnetic resonance imaging scans demonstrated a small residual lesion, which exhibited shrinkage in subsequent magnetic resonance imaging studies. The painful ticks disappeared and facial nerve weakness improved postoperatively. CONCLUSION: Although cerebellopontine angle fibromas may present similar radiological features, their clinical presentation may be somewhat different from that of typical VSs. If a fibroma is suspected, radiosurgery should be avoided; limited surgery may be considered as an option for patients experiencing symptoms. Because fibromas may be intraoperatively noted to be fibrotic and vascular, radical removal may not be easy or justified. After the final diagnosis has been reached, conservative treatment of the residual lesion may be the best option. PMID- 11063119 TI - Primary spinal epidural mantle cell lymphoma: case report. AB - OBJECTIVE AND IMPORTANCE: Mantle cell lymphoma is a distinct clinicopathological type of non-Hodgkin's lymphoma that often presents at an advanced stage, with systemic spread. Spinal involvement is uncommon and generally occurs as part of advanced disease or generalized relapses. Primary spinal epidural lymphoma is a rare initial manifestation of non-Hodgkin's lymphoma, and mantle cell lymphoma with initial presentation in the spinal epidural space is extremely rare, having been previously reported in only two cases. CLINICAL PRESENTATION: We report a case of a 71-year-old man who presented with increasing weakness and numbness of the legs. Magnetic resonance imaging revealed a spinal epidural mass in the lumbosacral region. INTERVENTION: The patient underwent a partial L4 and L5-S1 laminectomy, with incomplete resection of the mass for spinal decompression and tissue diagnosis. Mantle cell lymphoma was diagnosed in the pathological examination. CONCLUSION: After radiotherapy, the disease recurred with a soft tissue mass in the anterior maxillary area of the face. The patient underwent restaging and was treated with chemotherapy, with only a partial response. Mantle cell lymphoma with primary spinal epidural presentation is rare. This diagnosis can be established and other causes of spinal cord compression can be ruled out by obtaining tissue for proper histopathological examinations. Because of its aggressive behavior and poor prognosis, mantle cell lymphoma should be treated using a combined-modality approach. PMID- 11063120 TI - Nimodipine-induced acute hypoxemia: case report. AB - OBJECTIVE AND IMPORTANCE: Nimodipine is commonly used to improve neurological outcomes after subarachnoid hemorrhage. Although nimodipine reportedly has high specificity for the cerebral vasculature, adverse systemic effects such as hypotension have been described. This case report describes a patient with traumatic subarachnoid hemorrhage who experienced two episodes of previously undescribed, life-threatening hypoxemia that was directly related to nimodipine therapy. CLINICAL PRESENTATION: The patient experienced acute hypoxemia (partial pressures of oxygen of 32.9 and 58.7 mm Hg), on two separate occasions (3 d apart), that was temporally related to single doses of nimodipine therapy for traumatic subarachnoid hemorrhage. Other disease- and medication-related causes did not explain these episodes. INTERVENTION: After the inspired oxygen concentration was increased to 100% (both episodes) and the positive end expiratory pressure was increased to 7.5 mm Hg (first episode), the arterial oxygen saturation of the patient returned to baseline levels (>99%) within 40 minutes in each instance. Nimodipine therapy was discontinued after each episode. CONCLUSION: It is hypothesized that, in the presence of concomitant adult respiratory distress syndrome, nimodipine increased ventilation/perfusion ratio mismatch, through its direct vasodilatory effects on the pulmonary artery, and possibly interfered with the reflex hypoxic pulmonary vasoconstriction necessary to maintain adequate oxygenation for this patient. Clinicians should carefully monitor the oxygenation status of patients when nimodipine therapy is initiated. PMID- 11063121 TI - The Sick Lady, by Jan Steen (1626-1679). PMID- 11063122 TI - Balloon-assisted Guglielmi detachable coiling of wide-necked aneurysms: part I- experimental evaluation. PMID- 11063123 TI - A randomized, controlled study of a programmable shunt valve versus a conventional valve for patients with hydrocephalus. PMID- 11063124 TI - P16Ink4a tumor suppressor function in lung cancer cells involves cyclin-dependent kinase 2 inhibition by Cip/Kip protein redistribution. AB - As cell cycle regulators whose activity is frequently altered in human cancers, cyclin-dependent kinases (cdks) are novel targets for therapeutic intervention. cdk inhibition is an emerging strategy for the treatment of non-small cell lung carcinomas (NSCLCs) because most derived cell lines express functional retinoblastoma protein (Rb) but appear to bypass its function with inappropriate cdk activity. Elevated cdk4/cdk6 activity in NSCLC cells is often due to inactivation of the p16Ink4a cdk inhibitor. To model the effects of cdk4/cdk6 inhibition, we have expressed p16Ink4a in a Rb-positive NSCLC cell line that lacks endogenous p16Ink4a expression. Whereas cdk4/cdk6 inhibition and Rb dephosphorylation are expected on p16Ink4a expression, we have also observed indirect cdk2 inhibition. cdk2 inactivation by the redistribution of other cdk inhibitors may be required for p16Ink4a-mediated growth suppression of Rb positive cells. The implications of such a requirement on the use of chemical cdk inhibitors to treat human cancers will be discussed. PMID- 11063125 TI - Retinoic acid induces neuronal differentiation of embryonal carcinoma cells by reducing proteasome-dependent proteolysis of the cyclin-dependent inhibitor p27. AB - Retinoic acid (RA) treatment of embryonal carcinoma cell line NTERA-2 clone D1 (NT2/D1) induces growth arrest and terminal differentiation along the neuronal pathway. In the present study, we provide a functional link between RA and p27 function in the control of neuronal differentiation in NT2/D1 cells. We report that RA enhances p27 expression, which results in increased association with cyclin E/cyclin-dependent kinase 2 complexes and suppression of their activity; however, antisense clones, which have greatly reduced RA-dependent p27 inducibility (NT2-p27AS), continue to synthesize DNA and are unable to differentiate properly in response to RA as determined by lack of neurite outgrowth and by the failure to modify surface antigens. As to the mechanism involved in RA-dependent p27 upregulation, our data support the concept that RA reduces p27 protein degradation through the ubiquitin/proteasome-dependent pathway. Taken together, these findings demonstrate that in embryonal carcinoma cells, p27 expression is required for growth arrest and proper neuronal differentiation. PMID- 11063126 TI - Expression of E6 and E7 papillomavirus oncogenes in the outer root sheath of hair follicles extends the growth phase and bypasses resting at telogen. AB - Hair follicle growth cycle proceeds through a series of stages in which strict control of cell proliferation, differentiation, and cell death occurs. Transgenic mice expressing human papillomavirus type 16 E6/E7 papillomavirus oncogenes in the outer root sheath (ORS) display a fur phenotype characterized by lower hair density and the ability to regenerate hair much faster than wild-type mice. Regenerating hair follicles of transgenic mice show a longer growth phase (anagen), and although bulb regression (catagen) occurs, rest at telogen was not observed. No abnormalities were detected during the first cycle of hair follicle growth, but by the second cycle, initiation of catagen was delayed, and rest at telogen was again not attained, even in the presence of estradiol, a telogen resting signal. In conclusion, expression of E6/E7 in the ORS delays entrance to catagen and makes cells of the ORS insensitive to telogen resting signals bearing to a continuous hair follicle cycling in transgenic mice. PMID- 11063127 TI - Okadaic acid-mediated induction of the c-fos gene in estrogen receptor-negative human breast carcinoma cells utilized, in part, posttranscriptional mechanisms involving adenosine-uridine-rich elements. AB - Signal transduction via modulation of phosphorylation after selective inhibition of protein phosphatase (PP) 1 and/or PP2A appears to play a role in okadaic acid (OA)-mediated effects. Treatment of several estrogen receptor-negative human breast carcinoma (HBC) cells with 100 nM OA resulted in induction of c-fos, c myc, and cyclin-dependent kinase inhibitor p21WAF1/CIP1 genes. Transfections of various luciferase reporter constructs in HBC cells revealed involvement of activator protein-1-dependent as well as -independent pathways in induction of the c-fos gene by OA. MDA-MB-468 HBC cells were stably transfected with plasmids expressing luciferase, chimeric luciferase- c-fos 3' untranslated region (3'UTR), or chimeric luciferase-p21WAF1/CIP 3'UTR mRNAs. Expression of chimeric luciferase c-fos and luciferase-p21WAF1/CIP1 mRNAs was elevated by OA in several independent sublines. Actinomycin D chase experiments revealed an enhanced rate of decay of luciferase-c-fos mRNA, whereas treatment with OA caused approximately 3.5-fold enhanced stability of the chimeric luciferase-c-fos mRNA only. By transfecting different plasmids containing deletions of c-fos 3'UTR, OA-responsive sequences were mapped to an 86-nucleotide, AU-rich region. UV cross-linking experiments using HBC cell cytosolic proteins showed multiple complexes with the AU-rich region subfragments of c-fos, as well as c-myc and p21WAF1/CIP1 mRNAs. OA enhanced binding of a novel Mr approximately 75,000 protein present in the cytosolic extracts of HBC cells to the AU-rich RNA probes of all of the above three genes. Taken together, OA regulation of HBC cell gene expression involves the activator protein-1 pathway, as well as enhanced binding of a novel Mr approximately 75,000 protein to an AU-rich region of the 3'UTRs of the target genes. PMID- 11063128 TI - Expression of an alternative Dnmt1 isoform during muscle differentiation. AB - The methylation pattern of genomic DNA undergoes dramatic changes during mammalian development, with extensive de novo methylation occurring during gametogenesis and after implantation. We identified an alternative Dnmt1 transcript in skeletal muscle by Northern blot analysis and cloned the corresponding cDNA by rapid amplification of cDNA ends and reverse transcription PCR. Using an in vitro skeletal muscle differentiation system, we show that this alternative Dnmt1 isoform is specifically expressed in differentiated myotubes, whereas the ubiquitously expressed isoform is down-regulated during myogenesis. Sequence analysis showed that this skeletal Dnmt1 isoform is identical to the one present in testis, which had been described as untranslatable. Here we present evidence that this alternative Dnmt1 transcript present in testis and skeletal muscle is translated despite the presence of several out-of-frame upstream ATGs and gives rise to a shorter Dnmt1 isoform, which could play an active role in the change of DNA methylation patterns during gametogenesis and myogenesis. PMID- 11063129 TI - Cell cycle checkpoints and their inactivation in human cancer. AB - Checkpoints are mechanisms that regulate progression through the cell cycle insuring that each step takes place only once and in the right sequence. Mutations of checkpoint proteins are frequent in all types of cancer as defects in cell cycle control can lead to genetic instability. This review will focus on three major areas of cell cycle transition control, with particular attention to the alterations found in human cancer. These areas include the G1/S transition, where most cancer-related defects occur, the G2/M checkpoint and its activation in response to DNA damage, and the spindle checkpoint. PMID- 11063130 TI - Influence on antiproliferative activity of structural modification and conjugation of gonadotropin-releasing hormone (GnRH) analogues. AB - The effect of various GnRH analogues, and their conjugates on proliferation, clonogenicity and cell cycle phase distribution of MCF-7 and Ishikawa human cancer cell lines was studied. GnRH-III, a sea lamprey GnRH analogue reduced cell proliferation by 35% and clonogenicity by 55%. Structural modifications either decreased, or did not alter biological activity. Conjugation of GnRH analogues including MI-1544, MI-1892, and GnRH-III with poly(N-vinylpyrrolidone-co-maleic acid) (P) through a tetrapeptide spacer GFLG(X) substantially increased the inhibitory effect of the GnRH analogues. The conjugate P-X-GnRH-III induced significant accumulation of cells in the G2/M phase; from 8% to 15.6% at 24 h and 9.8% to 15% at 48 h. It was concluded that conjugation of various GnRH analogues substantially enhanced their antiproliferative activity, strongly reduced cell clonogenicity and retarded cell progression through the cell division cycle at the G2/M phase. PMID- 11063131 TI - Nonregenerative stimulation of hepatocyte proliferation in the rat: variable effects in relation to spontaneous liver growth; a possible link with metabolic induction. AB - Three procedures were used to stimulate hepatocyte proliferation in the rat without reducing liver mass, resulting in a supplementary growth which differs from the regenerative growth observed after loss of liver mass by hepatectomy or toxic necrosis. They were: (a) the ingestion of cyproterone, a cytochrome P450 inducing drug (b) the injection of an irritant which provokes glycogenesis and synthesis of acute-phase proteins (c) the injection of albumin-bound bilirubin leading to elimination of glucuronated bilirubin in bile. This ensuing supplementary growth was studied in the rat under several conditions of hepatic proliferation: 1. In normal adult rats, in which hepatocyte proliferation is very low, the effect on proliferation was either weak or undetectable. 2. In suckling rats, with a rapid body and liver growth, all the stimulants provoked a synchronized wave of proliferation with a steep increase of the percentage of S phase hepatocytes from 4.5% in controls to 15-30% in treated rats. This increase was followed by a compensatory period of low proliferation during which a treatment with a second stimulant was much less effective. 3. In 2/3 hepatectomized adult rats, the proliferation induced by cyproterone was higher than the spontaneous regenerative proliferation alone and additional to it during all of the regenerative process. The proliferation induced by acute inflammation was competitive with the synchronous spontaneous proliferation during the early period of synchronized proliferation following surgery, suggesting that both are similar acute responses. Differently, during the late period of lower and unsynchronized regenerative proliferation, the proliferation provoked by acute inflammation was additional to the spontaneous one. A stimulation of proliferation by injection of the albumin-bilirubin complex was observed during the late period after 2/3 hepatectomy. The highest level of stimulation occurred when the liver growth and the hepatocyte proliferation were already high. This suggests that these stimulants are not complete mitogenic stimuli and need cofactors which are present during the spontaneous growth or, alternatively, that the effect of stimulants is opposed by an inhibitory mechanism present in the adult rat. PMID- 11063132 TI - Gadd45 mutations are uncommon in human tumour cell lines. AB - GADD45 is an evolutionarily conserved gene that encodes a small acidic, nuclear protein and is an example of a p53 responsive gene. Gadd45 protein has been shown to interact with PCNA and also p21waf1. It has been implicated in growth arrest, DNA repair, chromatin structure and signal transduction. The confusing biochemical data has been clarified by the demonstration that Gadd45 null mice have a phenotype strikingly similar to that of p53 null mice, being tumour prone and showing marked genomic instability. We have tested the hypothesis that mutations in the GADD45 coding region might substitute for p53 abnormalities in tumour cell lines where p53 is wild type. After generating cDNA from mRNA in a panel of 24 cell lines we sequenced the GADD45 cDNA and have demonstrated that no mutations can be observed, even in the p53 wild type cell lines. Such data suggest that Gadd45 mutations are uncommon in human cancer. From this we postulate that, despite the phenotype of the GADD45 null mouse, GADD45 is unlikely to be the key mechanistic determinant of the tumour suppressor activity of the p53 pathway. PMID- 11063133 TI - Antisense oligonucleotides targeted against protein kinase c alpha inhibit proliferation of cultured avian myoblasts. AB - Protein kinase C (PKC) has been implicated in the control of proliferation and differentiation of many cell types. There is evidence indicating that it plays a role in signal transduction mechanisms related to myogenesis, but little is known about the individual functions of PKC isoforms in muscle cell development. Data obtained in previous studies using cultured chick embryo skeletal muscle cells suggested that PKC alpha is linked to the regulation of myoblast proliferation. However, this causal relationship could not be definitively established as no experiments based on selective inhibition of this isoform were carried out. In the present work, specific inhibition of the expression of PKC alpha in cultured myoblasts by using antisense oligonucleotide technology resulted in a significant decrease of culture cell density and DNA synthesis, clearly showing that this isoenzyme is involved in signalling pathways which promote muscle cell proliferation. PMID- 11063134 TI - A quantitative model for differential motility of gliomas in grey and white matter. AB - We have extended a mathematical model of gliomas based on proliferation and diffusion rates to incorporate the effects of augmented cell motility in white matter as compared to grey matter. Using a detailed mapping of the white and grey matter in the brain developed for a MRI simulator, we have been able to simulate model tumours on an anatomically accurate brain domain. Our simulations show good agreement with clinically observed tumour geometries and suggest paths of submicroscopic tumour invasion not detectable on CT or MRI images. We expect this model to give insight into microscopic and submicroscopic invasion of the human brain by glioma cells. This method gives insight in microscopic and submicroscopic invasion of the human brain by glioma cells. Additionally, the model can be useful in defining expected pathways of invasion by glioma cells and thereby identify regions of the brain on which to focus treatments. PMID- 11063135 TI - MCF-7 mammary tumour cells express the myeloid cell differentiation controlling factor, serine protease 3/myeloblastin. AB - Our previous data indicated that HSP27 plays a role in MCF-7 cell differentiation similar to that it has in HL-60 cells. In the latter case, this involves a control of its levels by proteinase 3/myeloblastin (PR3/Mbn), a serine proteinase hitherto considered specific of the myeloid lineage. Having observed that the treatment of MCF-7 cells with the serine protease inhibitor N-tosyl-l phenylalanine-chloromethyl ketone (TPCK) increased their content in HSP27 and induced them to acquire a secretory phenotype, we undertook this work to test the assumption that an enzyme similar or identical to PR3/Mbn might be expressed in this cell line. The data show that MCF-7 cells exhibited specific immunopositivity for a monoclonal antibody against PR3/Mbn. Western blot analysis of immunoprecipitates from MCF-7 cell extracts, obtained and checked with PR3/Mbn monoclonal antibodies, confirmed the presence of the 35 kDa glycosylated and 29 kDa mature forms of the protein. Finally, Northern blot analysis confirmed the expression of the corresponding mRNA. Together with our data with TPCK, this substantiates our hypothesis that, as in HL-60 cells, regulation of MCF-7 cells differentiation might involve a postranslation control on HSP27 levels by a serine protease. PMID- 11063136 TI - Peritoneal dialysis compared with hemodialysis in the treatment of end-stage renal disease. AB - Whether to use peritoneal dialysis (PD) or hemodialysis (HD) is a major decision in terms of clinical outcome and management implications; the final choice is difficult because of the conflicting results of comparisons reported in the literature. A review of studies comparing survival shows either superiority of HD, or superiority of PD, or equivalence of the two techniques, but an analysis of the comparisons as a whole brings to light two clear phases in the survival curves. In the first, residual renal function (RRF) gives PD an advantage, or at least puts it on the same level as HD. In the second phase, the reduction in Kt/V as RRF declines gives PD a potential risk. After a few years of PD treatment a sharp watch is therefore necessary to detect signs of under-dialysis promptly and to shift the patient to HD. In patients without RRF it is more difficult to control hypertension with PD and they are more prone to hyperhydration. Despite a widespread belief in the Eighties that PD was the treatment modality of election for diabetics, HD is in fact preferable in these patients, except younger ones. High-turnover and low-turnover bone lesions are more frequent respectively in HD and PD patients. Anemia is better controlled with PD. Blood lipids and nutritional indices are less well controlled with PD. Despite poor technical survival, the "pool" of patients treated with PD frequently reaches 20-30% because it is indicated as first treatment in a large proportion. PD preserves renal function better than HD and is useful while awaiting renal transplantation, with faster postoperative restoration of diuresis. The quality of life with PD as home treatment is usually better than with HD. In conclusion, dialytic centers should establish an integrated PD/HD programme as the two methods are not competitive but are different tools for the treatment and rehabilitation of uremic patients. PMID- 11063137 TI - Diuretic therapy and the risk for renal cell carcinoma. AB - Diuretics are important drugs in the treatment of arterial hypertension, congestive heart failure, the nephrotic syndrome, and other clinical conditions. With diuretic treatment, reduction of both morbidity and mortality has been clearly shown in those conditions. However, a recent meta-analysis showed an increased risk for renal cell carcinoma in patients treated with diuretics. We summarize and critically review data from studies examining the association between diuretic use and renal cell carcinoma. In general, in many clinical conditions, the reduction of mortality with diuretic therapy outweighs a potential risk of renal cell carcinoma. However, in certain conditions, alternative medical treatment should be considered. PMID- 11063138 TI - Apheresis in primary focal segmental glomerulosclerosis of native and transplanted kidneys: a therapeutic protocol. AB - BACKGROUND: Patients with focal segmental glomerulosclerosis (FSGS) develop nephrotic syndrome and terminal renal failure in most cases. FSGS reappears in 15 50% of transplanted kidneys and frequently causes the graft loss. Sera from patients with FSGS of native or transplanted kidneys contain some proteinuric or permeability factors (PF) which can be removed by means of plasma exchange (PE) or protein A Immunoadsorption (IA). METHODS: We suggest a therapeutic protocol, for patients with biopsy proven FSGS of native or transplanted kidneys, resistant to steroid and immunosuppressive therapy, based on the association of PE or IA to conventional drug therapy. Daily proteinuria, renal function, serum albumin and circulating level of proteinuric factors (permeability test) will be monitored at regular time intervals during the apheresis cycle, which will be intensive at the beginning (8-10 sessions in 4 weeks) and very gradually discontinued. Results. We will consider satisfactory remission the reduction of proteinuria below 1 g/day, improvement of renal function, normalization of serum albumin level (> 3.5 g/dl). Partial remission will be considered: proteinuria below 3 g/day, stable renal function, serum albumin level between 3 and 3.5 g/dl. Permeability test, if positive at baseline examination, should be negative after apheresis. CONCLUSIONS: The primary endpoint of our protocol is: lasting remission (satisfactory or partial) after the apheresis suspension. Secondary endpoints are: maintained remission with continuing apheresis sessions, correlation between permeability activity and disease activity, identification of responders and non responders patients on the basis of positive permeability test. PMID- 11063139 TI - Extrahepatic immunological manifestations of hepatitis C virus in dialysis patients. AB - BACKGROUND: Hepatitis C virus (HCV) infection may be associated with various extrahepatic immunological disorders. Uremic patients on chronic regular dialytic treatment (RDT) frequently develop immunological abnormalities. The aim of this study was to evaluate the probability that HCV infection creates an increased risk for extrahepatic immunological abnormalities in chronic RDT patients. SUBJECTS AND METHODS: In a series of one hundred sixteen chronic RDT patients, HCV status was determined by anti-HCV antibodies, polymerase chain reaction (PCR) RNA and viral genotyping. After excluding four anti-HCV negative/PCRRNA positive patients, a comparison was made between 51 anti-HCV negative/PCR-RNA negative and 61 anti-HCV positive patients, this latter group including seventeen PCR-RNA negative, fifteen genotype 1, thirteen genotype 2, three genotype 3, four genotype 4, four undeterminable genotype and five mixed genotypes. The following investigations were performed: cryoglobulinemia (presence, titer and, when possible, identification), monoclonal gammopathy, antineutrophil cytoplasm antibodies, antidouble stranded DNA antibodies, circulating immunocomplexes and immunoglobulin levels. RESULTS: Cryoglobulinemia was found in 77% of anti-HCV positive versus 29% of anti-HCV negative patients, and cryocrit > 1% in 50% versus 9.8% respectively, p=<0.01. Also cryoglobulin concentration was higher (logarithmic transformation: 4.38 +/- 0.94 vs 3.11 +/- 1.06, p =< 0.001) in anti HCV positive versus negative patients. Multivariate logistic regression analysis showed a significantly increased odds ratio (12.0, confidence interval 3.0 to 48.3) for having high levels of cryoglobulins (cryocrit >1%) after adjusting for age and dialytic age. The prevalence of this abnormality did not differ significantly among patients infected with different genotypes, but a tendency towards a lower frequency was observed in the anti-HCV positive/PCR negative subgroup. Cryoglobulins were identified as type I (2 anti-HCV positive case), type II (2 anti-HCV positive and 1 anti-HCV negative case) and type 3 (1 anti-HCV negative case). The frequency of monoclonal gammopathy was not significantly different between anti-HCV positive and anti-HCV negative patients (6.5% versus 2%) as well as that of the other parameters evaluated except for IgG concentration which was higher in the anti-HCV positive group (1,685 +/-605 versus 1349 +/- 352 mg/dl, p 0.006). Five events, potentially linked to HCV infection, occurred in our anti-HCV positive patients: 2 cases of porphyria cutanea, 1 case of unexplained peripheral neuropathy, 1 cutaneous leukocytoclastic vasculitis, 1 death for non-Hodgkin's lymphoma. In one anti-HCV positive patient treated with interferon-alpha, the presence of cryoglobulins, monoclonal gammopathy and high IgG levels strictly paralleled that of viremia, disappearing during the recovery phase under treatment and reappearing shortly after stopping treatment. CONCLUSIONS: HCV infection provides a significantly increased risk for developing extrahepatic immunological abnormalities also in chronic RDT patients. It is possible that the clinical relevance of this event might be scant because of the low level of these abnormalities, but an awareness of its possibility should to be taken into account. PMID- 11063140 TI - Treatment of vasculitic IgA nephropathy. AB - BACKGROUND: Patients with IgA nephropathy and histological vasculitic/crescentic lesions have a poor prognosis. We performed a retrospective study to assess whether treatment with steroids and immunosuppressants would preserve renal function by healing these lesions and thereby prevent progression to glomerular sclerosis and renal failure. METHODS: Sixteen patients with IgA nephropathy and a vasculitic/crescentic glomerulonephritis diagnosed by renal histology were treated with a reducing course of prednisolone (initial dose 60 mg/day). Six patients also received cyclophosphamide (2 mg/kg/day) for three months followed by azathioprine (100 mg/day) in five patients. Ten patients received azathioprine (100 mg/day) in addition to prednisolone. The median duration of treatment was 12 months (range 5-30 months). At the end of treatment each patient had a second renal biopsy. RESULTS: Following treatment there was a significant reduction in the proportion of glomeruli with acute vasculitic lesions from a median of 17.4% (range 4.8-57.5%) to 0 (range 0-15.8%) (p=0.001). There was an increase in the proportion of globally sclerosed glomeruli from a median of 13.4% (range 0-44.4%) to 21.5% (range 0-90%) after treatment but this did not significantly differ from baseline (p=0.24). The proportion of renal cortex with chronic tubular atrophy increased from 2.55% (0.4-57.7%) to 11.3% (0.3-61%) (p=0.09). The median duration of follow-up was 30 months (inter-quartile range 6-30 months). At both 12 and 24 months there was no significant increase in serum creatinine. Four patients, however, developed end-stage renal failure between 24 and 81 months. CONCLUSION: In this retrospective study we show that treatment with steroids and immunosuppressants leads to healing of vasculitic lesions and may thus arrest progression of glomerular scarring. PMID- 11063141 TI - Primary renal lymphoma does exist: case report and review of the literature. AB - Primary renal lymphoma (PRL) is a controversial and rare disease and there is still no agreement on its existence. Many cases have been reported in the literature, but clear diagnostic criteria have not yet been established. Most of the reported cases are questionable because of incomplete staging or the presence of extrarenal disease. Here we report a new case and a review of the literature based on a critical examination of the diagnostic procedure. Thus, probably only 29 cases, ours included, should be recognized as PRL, because only these cases fulfil the three diagnostic criteria and underwent complete diagnostic screening, including renal biopsy, bone marrow biopsy and thoraco-abdominal computerised tomography (CT). PMID- 11063142 TI - Tubulointerstitial nephritis and asymptomatic uveitis. AB - We describe a case of a 10 year-old boy who had fever, weakness, anorexia, weight loss and general malaise. No other remarkable symptoms were present. He had been treated with Aspirin and Ibuprofen. Deterioration of renal function, glucosuria, proteinuria, anemia and increased erythrocyte sedimentation rate were detected. After 7 days observation with no treatment, renal function worsened, glucosuria increased and fever persisted. A renal biopsy was performed and acute tubulointerstitial nephritis was diagnosed. The most common aetiologies of this entity were excluded. An ophthalmologic study revealed bilateral anterior uveitis, therefore the patient was diagnosed as having tubulointerstitial nephritis with uveitis. The child improved on corticosteroid therapy, but uveitis relapsed when treatment was stopped. PMID- 11063143 TI - Acute renal failure due to idiopathic tubulo-intestinal nephritis and uveitis: "TINU syndrome". Case report and review of the literature. AB - Acute renal failure due to idiopathic tubulo-interstitial nephritis associated with bilateral uveitis (TINU syndrome) is a rare clinical event, contracted mainly by girls or women. Here we report the clinical follow-up regarding a 22 year-old woman with acute renal failure (creat. clearance 13.5 ml/min) due to idiopathic tubulo-interstitial nephritis documented by renal biopsy, after bilateral uveitis which healed with local prednisone. The clinical history and the clinical follow-up of our patient were typical of the TINU syndrome. We were able to exclude all diseases causing acute tubulo-interstitial nephritis such as systemic infection, hypersensitivity to drugs, Behcet's disease, Sjogren syndrome, sarcoidosis, systemic lupus or vasculitides. The patient recovered after systemic prednisone. PMID- 11063144 TI - Delayed onset of uveitis in TINU syndrome. AB - We report here the clinical features and outcomes of two patients who presented idiopathic tubulo-interstitial nephritis and uveitis syndrome (TINU syndrome) with ocular disease following the onset of nephropathy. The initial symptoms were renal impairment with asthenia, anorexia and weight loss. An increase in urinary beta2-microglobulin was noticed at the initial checkup in both patients. Renal biopsies showed interstitial cellular infiltration without granulomas or tubular atrophy. No glomerular and vascular alterations were seen and immunofluorescent staining was uniformly negative. Systemic steroid therapy was given and renal function returned to normal within three months. Anterior uveitis occurred in both patients eight months later and responded well to local steroid therapy. Renal involvement in TINU syndrome mostly has a favorable outcome. Despite the possibility of spontaneous regression, systemic steroids may be beneficial in reducing the development of interstitial fibrosis. PMID- 11063145 TI - Lupus nephritis. PMID- 11063146 TI - Possible clinical importance of the transformation of Helicobacter pylori into coccoid forms. PMID- 11063147 TI - Investigation of gastro-oesophageal reflux in patients with an intact stomach: is oesophageal bilirubin monitoring a useful addition to pH monitoring? AB - OBJECTIVES: Ambulatory bilirubin monitoring has helped to establish the role of duodenal contents in gastro-oesophageal reflux disease. This study aims to define the clinical role of oesophageal bilirubin monitoring in reflux patients with an intact stomach. METHODS: In total, 113 consecutive patients with reflux symptoms were prospectively studied using combined ambulatory oesophageal pH and bilirubin monitoring. Patients were categorized as follows: no pathological reflux, isolated acid reflux, isolated bilirubin reflux, combined acid and bilirubin reflux. RESULTS: Thirty-eight patients (33%) had no pathological reflux, 49 patients (44%) had combined pathological acid and bilirubin reflux, and 17 patients (15%) had isolated pathological acid reflux. Only nine patients (8%) had isolated pathological bilirubin reflux. In these nine, the extent of pathological bilirubin reflux was small (median total bilirubin exposure time 12.2%, range 6.5%-20.7%) and mucosal damage was minimal (five had grade 1 oesophagitis, four had a normal oesophagus). In one patient, symptoms were temporally related to acid reflux, and in none were symptoms temporally related to bilirubin reflux. CONCLUSIONS: Isolated oesophageal bilirubin reflux in patients with an intact stomach is uncommon. In these patients mucosal injury is minimal, and reflux symptoms are not related to bilirubin reflux episodes. Further work is needed to define the role for oesophageal bilirubin monitoring in the investigation of reflux disease in patients with an intact stomach. PMID- 11063148 TI - Effect of somatostatin on lower esophageal sphincter characteristics in man. AB - BACKGROUND: Somatostatin (SST) is known for its inhibitory effect on the gastrointestinal tract. Transient lower esophageal sphincter relaxations (TLESR), low or absent LES pressure (LESP) and swallow-induced LES relaxations are the most important reflux mechanisms. METHODS: We have studied the effect of somatostatin on lower esophageal sphincter (LES) characteristics in man. Nine healthy volunteers participated in four experiments performed in random order and double-blind during continuous infusion of somatostatin (250 microg/h) or saline (control) under fasting and postprandial conditions. Esophageal motility was measured with sleeve manometry combined with pH metry. RESULTS: Under fasting conditions LESP was not influenced by somatostatin. Ingestion of the carbohydrate meal significantly (P < 0.01) decreased LESP. During continuous somatostatin infusion the postprandial decrease in LESP did not occur; LESP was even significantly (P < 0.05) increased over basal levels. Somatostatin did not significantly influence TLESR frequency, neither under basal conditions, nor postprandially. The residual pressure during swallow-induced LES relaxation was significantly (P < 0.05) increased by somatostatin. CONCLUSION: In humans somatostatin prevents postprandial reduction in LESP, does not affect TLESR, but inhibits swallow-induced LES relaxation. PMID- 11063149 TI - Gastroprotective effect of histamine and acid secretion on ammonia-induced gastric lesions in rats. AB - BACKGROUND: Previous studies have shown that ammonia produced by Helicobacter pylori urease or administrated intragastrically exhibits a toxic effect on the gastric mucosa. In the present study we investigated the influence of histamine and gastric acid secretion on ammonia (NH4OH)-induced gastric lesions. METHODS: The gastric mucosa in rats was exposed to NH4OH (1.5 ml of 250 mM solution) under basal conditions, after administration of histamine (1 mg/kg), urea with urease, and ranitidine (40 mg/kg subcutaneously) given alone or in combination. We measured the area of gastric lesions, gastric blood flow (GBF), plasma gastrin concentration, DNA synthesis, gastric acid secretion and gastric luminal concentration of PGE2. RESULTS: Application of NH4OH resulted in the formation of acute gastric lesions. This effect was accompanied by a fall in GBF, a rise in gastric pH, and a reduction in mucosal DNA synthesis. Administration of histamine 30 min prior to NH4OH reduced the area of gastric lesions. This was accompanied by an increase in GBF, DNA synthesis, and prostaglandin E2 (PGE2) production. Ranitidine given prior to NH4OH enhanced gastric mucosa damage, and reduced GBF and DNA synthesis. This effect was accompanied by a reduction in gastric acid secretion. Ranitidine given prior to histamine abolished gastric acid secretion and the protective effect of histamine against NH4OH-induced damage; these effects were accompanied by a decrease in GBF, DNA synthesis, and concentration of PGE2. Pretreatment with 2% urea with urease given prior to NH4OH reduced NH4OH lesions. This effect was associated with an increase in gastric acid secretion, gastric generation of PGE2, GBF, and DNA synthesis. Ranitidine given prior to urea with urease inhibited gastric acid secretion and the gastroprotective effect of urea-urease gastroprotection. CONCLUSIONS: Histamine and gastric secretion exhibit a protective effect against ammonia-induced gastric lesions. This effect appears to depend upon the stimulation of gastric acid secretion and PGE2 production, and the improvement of gastric microcirculation. PMID- 11063150 TI - High prevalence of Helicobacter heilmannii-associated gastritis in a small, predominantly rural area: further evidence in support of a zoonosis? AB - BACKGROUND: Primary hosts of Helicobacter heilmannii are domestic animals--cats, dogs and pigs, but rarely is it detected in gastric biopsies from humans. We found H. heilmannii in gastric biopsies obtained from patients living in a predominantly rural area. METHODS: We evaluated geographic and demographic data from the area and calculated both the total prevalence and the prevalence in each community in this area. Chi-squared test and exploratory data analysis were used for statistical evaluation. Histologic and clinical data were recorded. RESULTS: Forty-three communities, mostly rural, were identified in the area, the size of which is about 200 km2. H. heilmannii was detected in 33 patients (prevalence 2%) living in 20 communities. The prevalence of H. heilmannii statistically significantly varied in those communities from 0.06%-1.1%. It was possible to infer that there is a negative correlation between prevalence and community size. Chronic active gastritis was diagnosed in all patients. The active inflammation became inactive after eradication of H. heilmannii via common anti-Helicobacter therapy. CONCLUSION: A high prevalence of H. heilmannii-associated gastritis in a small, predominantly rural area and statistically significant differences in prevalence from one community to the next, i.e. highest in the smallest village and lowest in a small town with a rather urban lifestyle, were found. In our opinion, these prevalence rates may be a consequence of transmission of the infection from domestic animals, because animal-human contact is generally more common in villages than in towns. PMID- 11063151 TI - Cure of Helicobacter pylori infection after failed primary treatment: one-center results from 120 patients. AB - BACKGROUND: Treatment with a proton pump inhibitor (PPI) and antimicrobials cures Helicobacter pylori infection in about 90% of patients. This is a retrospective overview of our studies aiming to cure the infection in all compliant patients with failed initial therapy. METHODS: We retreated 120 (19% of 644) H. pylori infected patients whose initial therapy had failed. The retreatments included (i) triple therapy (TT): colloidal bismuth subcitrate, metronidazole, amoxicillin (or tetracycline); (ii) quadruple therapy (QT): TT and a PPI; or (iii) high doses of both a PPI and clarithromycin combined with a further 1-3 individually selected antimicrobials. The eradication results were determined after 6-12 months. RESULTS: The 1st retreatment was successful in 70 of 120 patients. The 2nd retreatment cured 25 of the remaining 42 patients, the 3rd 13 of 17, and the 4th the last 4 patients. The cumulative eradication rate (ITT) was 93% (95% CI: 88.9% 97.9%; 8 patients withdrew after a failed 1st retreatment) and the rate was 100% in the remaining 112 patients who accepted several retreatments. The 1st retreatment with TT cured 23% (95% CI: 12%-34%) of 57 patients and QT 85% (95% CI: 74%-96%) of 41 patients who had initially undergone a failed metronidazole based treatment. All retreatments were well tolerated. CONCLUSIONS: In this study, high doses of a PPI and clarithromycin combined with 1-3 antimicrobials according to susceptibility data proved to be the best drug combination in the cure of H. pylori infection after failed primary treatment. Giving imidazole- and bismuth-based QT (without clarithromycin) as the first-line treatment of H. pylori infection ensures that the number of failures remains low. PMID- 11063152 TI - Personality traits predict treatment outcome with an antidepressant in patients with functional gastrointestinal disorder. AB - BACKGROUND: We investigated the relationship between personality traits and response to treatment with the tetracyclic antidepressant mianserin or placebo in patients with functional gastrointestinal disorder (FGD) without psychopathology. METHODS: Forty-eight patients completed the Buss-Durkee Hostility Inventory, Neuroticism Extroversion Openness -Personality Inventory (NEO-PI), and Eysenck Personality Questionnaire (EPQ), neuroticism + lie subscales, before they were consecutively allocated to a 7-week double-blind treatment study with mianserin or placebo. Treatment response to pain and target symptoms were recorded daily with the Visual Analogue Scale and Clinical Global Improvement Scale at every visit. RESULTS: A low level of neuroticism and little concealed aggressiveness predicted treatment outcome with the antidepressant drug mianserin in non psychiatric patients with FGD. Inversely, moderate to high neuroticism and marked concealed aggressiveness predicted poor response to treatment. These findings were most prominent in women. Personality traits were better predictors of treatment outcome than serotonergic sensitivity assessed with the fenfluramine test. CONCLUSION: Assessment of the personality traits negativism, irritability, aggression, and neuroticism may predict response to drug treatment of FGD even when serotonergic sensitivity is controlled for. If confirmed in future studies, the findings point towards a more differential psychopharmacologic treatment of FGD. PMID- 11063153 TI - Intolerance to cereals is not specific for coeliac disease. AB - BACKGROUND: Abdominal complaints after ingestion of cereals are not uncommon. We assessed how reliable such a history is as a marker for the presence of overt coeliac disease, and whether we should also take into account latent coeliac disease and cereal allergy. METHODS: The study group comprised 93 consecutive adults from health centres spontaneously reporting abdominal symptoms after consumption of cereals. Small bowel mucosal morphology, CD3+, alphabeta+ and gammadelta+ intraepithelial lymphocytes (IELs), HLA DQ alleles and serum IgA class endomysial (EmA), tissue transglutaminase (tTg) and gliadin (AGA) antibodies were determined. Skin prick and patch tests and serum radioallergosorbent tests for cereals were carried out. Thirty non-coeliac adults served as biopsy controls. RESULTS: Eight (9%) patients had coeliac disease and one mild partial villous atrophy. Altogether 17 had an increased density of gamma delta+ IELs without atrophy. However, only seven (8%) showed evidence of latent coeliac disease, i.e. both an increase in gammadelta+ IELs and the presence of coeliac disease-type HLA. One or more of the allergy tests for cereals was positive in 19; 9 adopted a gluten-free diet and abdominal symptoms were alleviated in all. In non-coeliac patients, serum EmA and tTg tests were negative in all, whereas AGA was seen in 40%. CONCLUSIONS: Intolerance to cereals is not a specific sign of overt or latent coeliac disease. All experimental dietary interventions before proper diagnosis of coeliac disease are therefore to be discouraged. Allergy to cereals, on the other hand, should be considered even in adults. PMID- 11063154 TI - Gastrointestinal symptoms rating scale in coeliac disease patients on wheat starch-based gluten-free diets. AB - BACKGROUND: A wheat starch-based gluten-free diet is widely adopted in the treatment of coeliac disease, even though the products contain trace amounts of gluten. The aim here was to establish whether such a diet sustains abdominal symptoms. METHODS: The Gastrointestinal Symptom Rating Scale (GSRS) was applied to 58 coeliac disease patients on gluten-free diets and 110 non-coeliac controls. An estimate was made of daily dietary fibre and wheat starch-derived gluten. Psychological well-being was evaluated by a structured interview. Twenty-three coeliac patients consented to small-bowel biopsy. RESULTS: The mean GSRS score in coeliac disease patients did not differ from that in control subjects. Poorer psychological well-being was associated with abdominal symptoms in coeliac patients, whereas the daily amount of wheat starch had no effect on GSRS score. Overall dietary compliance was good, and villous atrophy was found in only 2 out of 23 patients. The average fibre consumption, 13 g per day, was lower than recommended. CONCLUSIONS: Wheat starch-based gluten-free products are well tolerated in coeliac disease patients, provided that their diets are otherwise strict. PMID- 11063155 TI - Implications of serum pepsinogen I in early endoscopic diagnosis of gastric cancer and dysplasia. Helsinki Gastritis Study Group. AB - BACKGROUND AND AIMS: The risk of gastric cancer (GCA) is increased in atrophic gastritis. A low serum pepsinogen group I (SPGI) level is a good serologic indicator of atrophic gastritis of the gastric corpus and fundus, and can be used for diagnosis of subjects with atrophic gastritis and of increased risk for GCA. The present study was undertaken to investigate whether SPGI assay and a diagnostic gastroscopy could enable the diagnosis of GCA at an early stage. MATERIAL AND METHODS: The study was carried out as part of the Alpha-Tocopherol, Beta-Carotene Cancer prevention study (ATBC study) in Finland, in which 22,436 male smokers aged 50-69 years were screened by SPGI. Low SPGI levels (< 25 microg/l) were found in 2196 (9.8%) men. Upper GI endoscopy (gastroscopy) was performed in 1344 men (61%) and 78% of these had moderate or severe atrophic corpus gastritis in endoscopic biopsies. A control series of 136 men from the ATBC study cohort with abdominal symptoms, but with SPGI > or = 50 microg/l were similarly endoscopied, and 2.2% of these had corpus atrophy. RESULTS: Neoplastic alterations were found in 63 (4.7%; 95% CI: 3.6%-5.8%) of the 1344 endoscopied men with low SPGI levels. Of these, 42 were definite dysplasias of low grade, 7 dysplasias of high grade, 11 invasive carcinomas, of which 7 were 'early' cancers, and 3 carcinoid tumors. In the control series, 1 man (0.7%) of the 136 men had a definite low-grade dysplasia. Thus, 18 (1.3%; 95% CI 0.7%-2.0%) cases with 'severe' neoplastic lesions (4 advanced cancers, 7 early cancers and 7 dysplasias of high grade) were found in the low SPGI group, but there were none in the control group. All four patients with advanced cancer died from the malignancy within 5 years (mean survival time 2.5 years), whereas surgical treatment in all those with early cancer or high-grade dysplasia was curative. One of the seven patients with early cancer and two of the seven with high-grade dysplasia died within 5 years, but none died from the gastric cancer. Thus, curative treatment was given to 14 of 18 men in whom a malignant lesion was found in gastroscopy. This is about 15% of all gastric cancer cases (92 cases) which were diagnosed within 5 years after SPGI screening in the 22,436 men. Among the gastric cancer cases of the main ATBC study, the 5-year survival rate was 33% (85% of the non-survivors died from gastric cancer). CONCLUSIONS: We conclude that assay of SPGI followed by endoscopy is an approach which can enable the early diagnosis of gastric cancer at a curable stage. PMID- 11063156 TI - The effect of indomethacin on hepatitis B virus replication in chronic healthy carriers. AB - BACKGROUND: A chronic HBsAg carrier state, a major cause of viral spread in a community, is one of the consequences of hepatitis B virus (HBV) infection. Although successful immunization programs have been initiated to eliminate the virus, there is still a large number of people with HBV infection worldwide. This study was designed to investigate the effect of indomethacin treatment on HBV markers in humans, in comparison with a control group. METHODS: In total, 65 chronic 'healthy' HBV carriers were involved in the study. Patients were divided randomly into two groups. Group I (n = 42) received oral indomethacin 75 mg daily for 6 months. Group II (n = 23) acted as control. Patients in both groups were followed up for 6 months, during which laboratory tests, including viral parameters, were performed periodically. Liver biopsy was done in 17 patients (11/42 of the indomethacin group and 6/23 of the control group). RESULTS: All liver biopsies showed grade 0-2 and stage 0-1 HBV in both groups (P > 0.05). HBsAg positivity did not change in any patient in either group. Five patients who had positive HBeAg in group I became negative 4 months later, while patients in group II continued to be positive at 6 months (P < 0.001). Similarly, all patients receiving indomethacin exhibited a total anti-HBeAg immunoglobulin response at 6 months, while the control group remained the same during this period (P < 0.05). HBV DNA, as detected by polymerase chain reaction in 20/22 (91%), was negative in group I at the end of 6 months. No change was observed in group II (P = 0.007). CONCLUSIONS: Although no biochemical analyses were performed on prostaglandins in the present study, the results suggest that the prostaglandin pathway may be involved in the pathogenesis of the immune response against HBV, and that the suppression of viral replication is achieved as indicated by the disappearance of HBeAg and HBV DNA in healthy chronic HBV carriers. PMID- 11063157 TI - Mannose-binding lectin polymorphisms in patients with hepatitis C virus infection. AB - BACKGROUND: Persistent infection with hepatitis C virus (HCV) leads to liver cirrhosis (LC) and often to liver cancer. Little is known about host factors that determine the variable natural history. Mannose-binding lectin (MBL) is an important constituent of the innate immune system. In white patients there is an association between codon 52 mutation of the MBL gene and persistent hepatitis B virus (HBV) infection. To determine whether MBL gene polymorphisms affect the course of HCV infection, we investigated the association between MBL gene polymorphisms and HCV infection in Japanese subjects. METHODS: Fifty-two HCV infected Japanese patients (8 with chronic inactive hepatitis (CIH), 31 with chronic active hepatitis (CAH), 13 with LC) and 50 normal controls were studied. MBL gene mutations were determined by means of polymerase chain reaction and restriction fragment length polymorphism analyses. RESULTS: Codon 52 and codon 57 mutations were absent in all subjects. Homozygous mutation in codon 54 was present in one (0.9%) patient. Heterozygous codon 54 mutation was present in 17 (32%) of the 52 patients and in 21 (41%) of the controls. No significant difference in the frequency of codon 54 mutation was observed between patient and control groups. However, although no significant relationship was observed between MBL polymorphisms and the levels of HCV RNA, all patients with heterozygous or homozygous codon 54 mutations had CAH or LC. In contrast, 8 of the 34 patients without codon 54 mutation remained at CIH. (P = 0.0405). CONCLUSION: MBL may be one of the factors that influence the course of HCV infection. PMID- 11063158 TI - Apolipoprotein E alleles in women with intrahepatic cholestasis of pregnancy. AB - OBJECTIVE: To investigate the frequency of apolipoprotein E (apoE) alleles among women with intrahepatic cholestasis of pregnancy. METHODS: The presence of the three most common apoE alleles (epsilon2, epsilon3, epsilon4) was determined by polymerase chain reaction-restriction fragment length polymorphism in two groups of women: healthy pregnant women (n = 47) and pregnant women with a diagnosis of intrahepatic cholestasis of pregnancy (n = 44). In addition, the frequencies of the alleles in the general population in our area are presented for comparison. RESULTS: The frequency of the apo epsilon4 allele was 21.6% among women with intrahepatic cholestasis and it was 16.0% among healthy pregnant women (Fisher exact test; P= 0.216), which is close to the rate in the general population in our area (19%). None of the apoE genotypes was significantly over-represented, and homozygous genotype epsilon4 was not associated with more severe clinical disease than other genotypes. CONCLUSION: The observed profiles of allele and genotype frequencies confirm the equilibrium state between apoE polymorphism and obstetric cholestasis and suggest that apoE does not play a major role in the development of intrahepatic cholestasis of pregnancy. PMID- 11063159 TI - Experimental model of hepatic fibrosis following repeated periportal necrosis induced by allylalcohol. AB - BACKGROUND/AIMS: In most patients with chronic viral hepatitis the predominant lobular location of hepatic necrosis and fibrosis is the periportal zone. We established a new simple model of hepatic fibrosis in rats by repetitive periportal necrosis with allylalcohol. METHODS: Of 40 male adult rats, 30 were injected with 0.62 mmol/kg of allylalcohol intraperitoneally twice a week, the remaining 10 with normal saline as controls. Ten rats were killed at each of 4, 8, and 16 weeks later. The extent of fibrosis was evaluated according to the portal-portal extent. Transforming growth factor (TGF) beta1 mRNA in liver tissues was detected by reverse transcriptase polymerase chain reaction, and its levels were determined by the endpoint titers of serial two-fold dilutions of cDNA. RESULTS: After 4 weeks, periportal fibrosis was produced in only 6 out of 10 rats, and was mild in extent. However, after 8 weeks, 8 out of 9 survivors showed moderate to severe fibrosis, which corresponded to a score of 7 or more. The extent of fibrosis correlated significantly with the amount of collagen and TGFbeta1 mRNA expression in liver tissues. The collagen content and expression of TGFbeta1 mRNA were also upregulated significantly in liver tissues with a fibrosis score of 7 or more. CONCLUSIONS: Hepatic fibrosis can be sufficiently induced by repetitive intraperitoneal injection of 0.62 mmol/kg of allylalcohol twice a week for 8 weeks. This simple model of hepatic fibrosis, in which TGFbeta1 is overexpressed at the transcriptional level, may be useful in the study of patients who have predominantly periportal necrosis and fibrosis. PMID- 11063160 TI - Fatty liver in obesity: relation to Doppler perfusion index measurement of the liver. AB - BACKGROUND: The correlation of clinical and laboratory findings with various imaging techniques in obese patients is difficult. Colour duplex Doppler is of particularly limited value in fat individuals. The Doppler Perfusion Index (DPI) measures the ratio of hepatic arterial to total liver blood flow and seems to be more accurate in the study of hepatic hemodynamics. The aim of the present study was to investigate the clinical use of DPI measurement of the liver in obesity. METHODS: In the present prospective, open study we evaluated the DPI in 41 obese patients (body mass index (BMI) > 30 kg/m2) and 18 volunteers with normal or slightly increased weight. Thirty patients of the study group underwent liver biopsy during bariatric surgery. In these patients liver histology was assessed and age, BMI, waist to hip ratio (WHR), DPI, liver function tests and serum triglycerides were measured. RESULTS: Obese patients had significantly (P = 0.0036) higher DPI values (0.25 +/- 0.138) than the healthy volunteers (0.15 +/- 0.04). Multivariate analysis revealed that grade of fatty liver in the study group was inversely associated with DPI and positively depended on serum triglyceride and aspartate aminotransferase (ASAT) levels (fatty liver index = 1.03 x ASAT (IU/l) + 0.152 x triglyceride (mg%) - 49.75*DPI, with P < 0.0001 and r2 = 0.80). CONCLUSION: Grade of fatty liver in obese patients may be predicted from DPI, serum triglyceride and AST levels. The proposed index may be useful as a non-invasive diagnostic tool during the follow-up of patients with obesity related fatty liver. PMID- 11063161 TI - Detection of Epstein-Barr virus DNA in hepatocellular carcinoma tissues from hepatitis C-positive patients. AB - BACKGROUND: We have previously shown that hepatitis C virus (HCV) replication is promoted by the Epstein-Barr virus (EBV) in vitro. The aim of this study was to examine the EBV load in hepatocellular carcinoma (HCC) tissues from HCV antibody positive patients. METHODS: DNA was extracted from paraffin sections from 168 HCC patients. After amplification of a region in the EBV BamHI W sequence by means of the polymerase chain reaction (PCR), it was detected by Southern hybridization and semi-quantified. Ten hyperplastic lesions from HCV-positive patients and 35 non-tumorous samples from hepatitis-negative patients served as controls. The PCR results were analyzed on the basis of the patient's hepatitis status. Univariate and multivariate analyses were performed to identify clinicopathologic factors for predicting EBV infection in HCC tissues. RESULTS: More than one copy of EBV DNA per 100 cells was detected in 56 (33%) of the HCC sections. The detection ratio in HCC tissues from HCV antibody-positive patients was 40% (45 of 113), which was significantly higher than that in tissues from HBV surface antigen positive patients (14%, 5 of 37; P = 0.0018). The patient's serum HBV surface antigen and HCV antibody independently predicted the EBV positivity of HCC tissues. CONCLUSIONS: These results support our hypothesis that EBV could play an important role in the development of HCV-related HCC. PMID- 11063162 TI - Outcome of gastrointestinal complications after liver transplantation for familial amyloidotic polyneuropathy. AB - BACKGROUND: Gastrointestinal disturbances are important prognostic factors for mortality and morbidity after liver transplantation for familial amyloidotic polyneuropathy (FAP). However, the impact of liver transplantation on malabsorption and bacterial small-bowel contamination has not been evaluated. METHODS: Twenty-three FAP patients were available for the study. They were examined for gastrointestinal disturbances as a part of the evaluation for liver transplantation for FAP. Bile acid malabsorption was diagnosed with the [75Se] homocholic acid taurate (SeHCAT) test; fat malabsorption by measuring faecal fat excretion; and bacterial small-bowel contamination with the hydrogen breath test (HBT). RESULTS: No significant improvement of malabsorption test results were noted from the pre-transplant evaluation 8 months (range, 2-20 months) before transplantation to the post-transplant evaluation performed a median of 20 months (range, 9-62 months) after the procedure. The SeHCAT test result became abnormal in two patients and normal in one, and changes in the test correlated with the time the patients were waiting for transplantation. Faecal fat excretion after transplantation correlated with duration of the disease and with fat excretion before transplantation. A significantly increased fat excretion was noted at the post-transplant evaluation. A change in HBT result was noted in only one patient, in whom the test result became normal; pre-transplant values correlated with those obtained after transplantation. CONCLUSION: For most FAP patients no improvement in gastrointestinal function was found after transplantation. The finding underlines the importance of an early transplantation before the patients have developed gastrointestinal dysfunction. PMID- 11063163 TI - Effect of conscious sedation on cardiac autonomic regulation during colonoscopy. AB - BACKGROUND: Colonoscopy is associated with cardiovascular events including hypotension, hypertension, and myocardial ischemia. The pathogenetic mechanisms of these cardiovascular events are unknown, but there is evidence that the autonomic nervous system may play a role. Conscious sedation is often used to relieve the inconvenience caused by the procedure. In this study, we evaluated the effects of sedation on cardiac autonomic regulation during colonoscopy. METHODS: One hundred and eighty patients undergoing elective colonoscopy were prospectively randomized into three groups: (i) sedation with intravenous midazolam (midazolam group); (ii) sedation with intravenous saline (placebo group); and (iii) no intravenous cannula (control group). Continuous electrocardiogram was recorded prior to, during, and after the colonoscopic procedure. Heart rate variability (HRV) was assessed by means of the power spectral analysis; the powers of low-frequency (LF 0.04-0.15 Hz) and high frequency (HF 0.15-0.40 Hz) components were calculated. RESULTS: Intubation of the colonoscope increased the LF component of HRV and decreased HF power in all study groups compared to baseline recording. Furthermore, compared to baseline, the LF/HF ratio--a marker of cardiac sympathetic regulation--increased during intubation in the midazolam (P < 0.001) and placebo (P < 0.05) groups, with no change in the control group. During intubation the midazolam group presented with higher LF and lower HF power than placebo (P < 0.001) and control groups (P < 0.01). Accordingly, the LF/HF ratio was higher in the midazolam group than in the placebo (P < 0.05) or control groups (P < 0.05). CONCLUSIONS: Midazolam potentiates the dominance of the sympathetic nervous system induced by colonoscopy. Therefore, conscious sedation with midazolam may contribute to the occurrence of cardiovascular events during colonoscopy. PMID- 11063164 TI - Comparison of a linear miniature ultrasound probe and a radial-scanning echoendoscope in TN staging of esophageal cancer. AB - BACKGROUND: Endoscopic ultrasonography is a precise method for TN staging of esophageal cancer. We explored the staging properties of a linear miniprobe as compared with a radial-scanning echoendoscope. METHODS: Sixty-eight patients with esophageal cancer underwent preoperative TN staging using a 20-MHz linear miniprobe and a 7.5/12-MHz radial-scanning echoendoscope. Tumor stage was verified by surgery and/or histology. RESULTS: T and N stages were verified in 53 and 54 patients, respectively. T-staging accuracy using the echoendoscope was 70%. The high-frequency miniprobe could not differentiate between T3 and T4 tumors, but both systems had an accuracy of 87% in discriminating between T1, T2, and T3/4 stages. With traversable tumors, the accuracy of N staging was significantly better with the echoendoscope than with the miniprobe (90% vs. 48%, P = 0.008). CONCLUSIONS: The two endosonographic systems had similar accuracy for assessing transmural tumor growth, but the echoendoscope was superior in staging advanced transmural tumors and in predicting lymph node metastasis with traversable tumors. PMID- 11063165 TI - Extrapulmonary sarcoidosis primarily diagnosed in the liver. AB - Sarcoidosis is a relatively common, chronic, multisystem disease of unknown origin characterized by the presence of noncaseating epithelioid granulomas. Although an array of organs may be affected by the disease, the commonest site of affection is the lung. We describe a 73-year-old patient admitted to our hospital because of fatigue, weight loss, and an increased alkaline phosphatase level. In conjunction with clinical presentation, laboratory variables, and imaging analysis, a liver biopsy finally confirmed the diagnosis of a systemic sarcoidosis without affection of the lung or mediastinal lymph nodes. Treatment with ursodeoxycholic acid before diagnosis did not improve clinical symptoms and cholestasis indicators. After prednisone treatment, liver enzyme values normalized and remained normal during follow-up for 2 years after diagnosis. The literature on hepatic manifestation of sarcoidosis, its diagnosis, treatment, and prognosis is reviewed. This single case of sarcoidosis presented to the clinician almost exclusively with liver enzyme abnormalities. The consideration of sarcoidosis in such cases is of utmost importance, since the differential diagnosis of hepatic granulomas includes infectious diseases in which treatment with corticosteroids could be fatal. PMID- 11063166 TI - Acute life-threatening asthma: not a time to procrastinate! PMID- 11063167 TI - A community study of male androgenetic alopecia in Bishan, Singapore. AB - BACKGROUND: Androgenetic alopecia is the most common form of hair loss. It affects a large number of the local male population, with 1,812 men seeking treatment for hair loss at the sole dermatological tertiary referral centre in Singapore in 1994. The aim of this study was to assess the prevalence of male androgenetic alopecia in the community. METHODS: A questionnaire-based cross sectional survey with a one-stage sampling method was conducted. Each male was diagnosed clinically and the severity graded according to the Norwood Criteria. The survey area was in Bishan East, a housing estate with 8,004 households. A total of 335 households were selected for the survey. RESULTS: The household response rate was 84%. Within these households, 254 out of 378 men participated in the study (67% response rate). The prevalence of androgenetic alopecia was found to be 63%. The prevalence of the condition increased with age, from 32% among young adults aged 17 to 26 years to 100% among those in their 80s. Proportionately more Indians (87%) were affected compared to Chinese (61%). 81% of the respondents with androgenetic alopecia did not seek help as they did not view it as a problem. Of those seeking treatment, 74% used non-medical methods of unproven effectiveness. CONCLUSION: There is a high prevalence of androgenetic alopecia in the community in Singapore. Age specific prevalence and racial differences correlate well with both Western and local studies respectively. PMID- 11063168 TI - Laparoscopy in the evaluation of the non-palpable undescended testes. AB - AIM OF STUDY: To examine the role of laparoscopy in the evaluation of the non palpable undescended testes in paediatric patients. METHOD: A review of all laparoscopies performed for the evaluation of the non-palpable undescended testes in a children's hospital over a 12-month period was conducted. Special attention was paid to the patients' age, the location of the testes at the time of laparoscopy, the subsequent surgical procedures and the complications. RESULTS: Sixteen boys underwent laparoscopy to localise 20 nonpalpable testes. 12 patients had unilateral disease while 4 had bilateral disease. 15 non-palpable testes were intraabdominal, 3 had inguinal testicular remnants and 2 had 'vanished'. A contralateral inguinal hernia was incidentally found in one patient. A patient with epididymal-gonadal non-union was not apparent at laparoscopy. There were no post-laparoscopic complications in all 16 patients. CONCLUSION: Laparoscopy is safe and accurate in the evaluation of non-palpable testes in children. The accurate localisation of intraabdominal testes facilitates the development of an optimal surgical strategy. PMID- 11063169 TI - Botulinum toxin A in the treatment of hemiplegic spastic foot drop--clinical and functional outcomes. AB - PURPOSE OF STUDY: This study investigated the effects of intramuscular Botulinum toxin A (BTX-A) in 7 ambulatory chronic hemiplegic subjects (5 male, 2 female) who had spastic hemiplegic foot drop. BASIC PROCEDURES: An open label study involving intramuscular injections of Botulinum toxin A (dilution 10 U/0.1 ml) was performed in ambulatory chronic hemiplegics. Tone as measured by the Modified Ashworth Scale (MAS), passive ankle joint range of motion (PROM), briskness of ankle reflexes, gait velocity, motor functional status and effects on the use of walking aids were measured at baseline, 3 and 12 weeks post-injection. MAIN FINDINGS: All subjects except I showed a significant decrease in MAS from 3.43 +/ 0.54 at baseline to 2.0 +/- 1.15 at 3 weeks post-injection, which was maintained during the 3 month study duration. The median change in PROM was 17.0 degrees (SD 12.1 degrees) at 3 weeks and 5.0 degrees (SD 7.1 degrees) at 12 weeks (p = 0.25) Gait velocity and Modified Barthel Index mobility scores which measured motor functional status were not significantly altered post-injection. The injections were generally well-tolerated and there were no serious adverse side effects. PRINCIPAL CONCLUSIONS: Although significant decreases in muscle tone were observed and maintained after intramuscular Botulinum toxin A during the 3 month study period, this regional intervention did not significantly influence functional status, gait velocity and the use of ambulatory aids. PMID- 11063170 TI - Clinical features and outcome of management of severe acute asthma (status asthmaticus) in the intensive care unit of a tertiary medical center. AB - AIM OF THE STUDY: The recognition and management of severe acute asthma have attracted considerable attention since the seventies because of the morbidity and mortality that may accompany the condition. Recognition and appropriate management of severe acute asthma is essential. Admission to intensive care, intubation and ventilation risks versus benefit have been argued. We highlight these controversies by documenting our experience and comparing it to others in the literature METHODOLOGY: We prospectively document our experience over a two year period in the management of severe asthma in the intensive Care Unit (ICU). Patients were established asthmatics, who came in severe exacerbation. Attention was paid to the duration of onset of acute attack, time to presentation, spirometric and blood gas data, the type of treatment given, factors responsible for complications and mortality were identified. The findings in this study were compared with those in similar studies in the literature. RESULTS: A total of 30 patients were studied. Twenty-one patients were ventilated and 9 were not. 82% had a history of asthma longer than 5 years. The duration of symptoms before admission to ICU was very short (one day or less in 57%). Hypercapnia was significantly higher in intubated patients. The duration of stay in ICU and hospital was longer for intubated patients (P<0.02). Complications were higher in intubated patients. CONCLUSION: ICU care provides an excellent setting for management of acute severe asthma. The reported high morbidity and mortality in ICU can be improved. Without ICU care the mortality and morbidity increases,so physicians should not hesitate to admit asthmatics early to ICU. PMID- 11063171 TI - Adjusting to military life--servicemen with problems coping and their outcomes. AB - A small proportion of servicemen enlisting for compulsory National Service in Singapore experience problems adjusting to military life. This paper aims to profile the servicemen who experience such problems. There is a paucity of literature addressing this issue internationally and none published locally. Servicemen who were referred to the Psychological Medicine Branch of the Singapore Armed Forces within six months of enlistment were retrospectively studied. In the work year July 1995 to June 1996, 77 cases were seen. The main classes of diagnoses were stress-related disorders, anxiety, mood and psychotic disorders. The main stressor was problems adapting to the military environment. There were 10 cases of parasuicide, significantly less than US Army statistics. At Operationally Ready Date, 20.7% were able to hold a combat vocation, similar to the US Army situation. This paper hopes to document the local figures and act as a reference for evaluating future therapies and policies. PMID- 11063172 TI - A case report of mycobacterium marinum infection of the hand. AB - We report a case of Mycobacterium marinum infection of the hand presenting initially as triggering of the digits. We like to highlight the unusual source of the infection and difficulty of diagnosis in this case as well as the various treatment modalities. PMID- 11063173 TI - The increasing prevalence of Endemic Typhus in Kuala Lumpur and an evaluation of a diagnostic ELISA dot test for the detection of antibodies to Rickettsia typhi. AB - A seroepidemiology study was done in response to the recent increase of Endemic Typhus cases diagnosed at University Hospital. The serosurvey was based on doctors' request for the Weil Felix (WF) or the Indirect Immunoperoxidase (IIP) test in Pyrexia of Unknown Origin (PUO) patients for the years 1991 to 1997. Over the 7 years, we found that the incidence of Endemic typhus is increasing with gender (male:female = 2:1), age (20-40 years) and race distribution (Indians > Malay > Chinese) that reflects socioeconomic circumstances. A commercially available ELISA dot assay [INDX (E2R3) Dip-S-Ticks], for the detection of antibodies against R. typhi was compared with the indirect immunoperoxidase test (IIP). The ELISA assay was done against 219 IIP tested sera. The Dip-S-Ticks was found to be comparable to the IIP with a sensitivity of 91.7% and specificity of 92.8% at cut-off titres of > 1:80 IIP. PMID- 11063174 TI - Severe eosinophilia in disseminated gastric carcinoma. AB - Peripheral blood eosinophilia is a well-recognised but unusual manifestation of malignancy, and may represent a paraneoplastic phenomenon. We present a case of poorly differentiated adenocarcinoma of the stomach associated with severe peripheral blood eosinophilia A 55-year old man was admitted for abdominal pain of one week duration. An incidental finding of leucocytosis with eosinophilia was noted. After excluding haematological and infectious causes, an oesophagogastroduodenoscopy (OGD) followed by biopsy confirmed the diagnosis. Eosinophilia appears to be a response to cytokine production,and treatment is aimed at the underlying malignancy, and reducing the eosinophil count when necessary, to prevent end-organ damage. Studies have shown that peripheral eosinophilia is associated with disseminated, metastatic disease and hence signifies a poor prognosis,whereas tissue eosinophilia in advanced cancer has a better survival rate. PMID- 11063175 TI - Hypertension in the young adult--come feel the pulse. AB - Hypertension occurring in teenagers and young adults is uncommon. Though the most common form is still essential hypertension, secondary causes are more commonly found here than in older adults. Renal, cardiovascular and endocrine diseases constitute most of these causes. Coarctation of the aorta is the most common cardiovascular cause of hypertension, and its importance lies in the fact that it is correctable, and that its persistence often leads to dangerous complications and early death. The cardinal sign of differential pulse and blood pressures between the upper and lower limbs can be detected clinically. Hence, the importance of a detailed physical examination in all young hypertensives, including palpation of all the pulses, cannot be overemphasized. We present 2 hypertensive young men who were found to have isolated coarctation of the aorta. The lesion in the first patient was located postductally just distal to the left subclavian artery. This area has been found to be the most common site of coarctation. The second patient had an unusual mid-thoracic coarctation. The clinical and radiological features as well as complications are highlighted. In young hypertensive patients, a high index of suspicion may enable the physician to make a timely diagnosis and hence avert the potentially disastrous complications that may arise in undetected cases. PMID- 11063176 TI - Clinics in diagnostic imaging (49). Cerebello-tonsillar haemangioblastoma. AB - A 39-year-old man with cerebello-tonsillar haemangioblastoma is presented,with emphasis on its morphological and imaging features. Although both computed tomography and cerebral angiography are very useful imaging modalities in helping to establish a radiological diagnosis, magnetic resonance (MR) imaging is more accurate and sensitive in depicting the lesion nature and extent. MR imaging is especially reliable in detecting multiple and other associated lesions within the central nervous system, such that a conclusive diagnosis of the von Hippel-Lindau syndrome can be made in selected cases. It is important to identify isolated haemangioblastomas as they constitute a relatively high percentage of benign resectable tumours within the posterior fossa.The differential diagnosis and imaging features of other posterior fossa tumours are also discussed. PMID- 11063177 TI - A comparison of commonly used anti-emetics for the prevention of emetic sequelae after a major gynaecological surgery. AB - STUDY DESIGN: Prospective, randomized, double-blind, placebo-controlled study involving one hundred ASA I-II patients undergoing major gynaecological surgery. OBJECTIVE: To study anti-emetic efficacy of intravenous (IV) ondansetron (4 mg), droperidol (2.5 mg), metoclopramide (10 mg), and placebo. PATIENTS AND METHODS: 100ASA physical status I-II undergoing major gynaecological surgery were randomized to receive intravenously (IV), one of the four test drugs 10 minutes before the end of anaesthesia. The incidence of postoperative nausea and vomiting following a standard anaesthetic technique was assessed. RESULTS: A significantly large number of patients who received ondansetron (88%) and droperidol (72%) were free of emetic sequelae when compared to placebo (41%); p < 0.05 (power of this observation is approximately 80% at the given significance level). Metoclopramide was ineffective. Patients given droperidol were significantly more sedated than those receiving ondansetron; p < 0.05. This is not surprising, as the dose of droperidol used in this study was higher than that currently recommended because we found lower doses to be ineffective in controlling nausea and vomiting in this group of patients. CONCLUSION: It was concluded that, of the drugs studied ondansetron is the best choice for anti-emetic prophylaxis after major gynaecological surgery. PMID- 11063178 TI - Seroprevalence of cytomegalovirus, toxoplasma and parvovirus in pregnancy. AB - AIM OF STUDY: The aim of our study was to determine the seroprevalence of cytomegalovirus (CMV), toxoplasma and parvovirus infection in our local antenatal population, and to see the effects, if any, of age, race, parity and nationality on its seroprevalence. METHODOLOGY: The sera of 120 consecutive antenatal women seen in KK Women's and Children's Hospital between the period of October 1997 and March 1998 were screened for cytomegalovirus (CMV) IgG, toxoplasma IgG and parvovirus B19 IgG and IgM. An antibody titer greater than 1:32 was regarded as positive. RESULTS: A total of 87.0% of patients were tested seropositive for CMV IgG, 17.2% seropositive for toxoplasma IgG and 30.0% seropositive for parvovirus IgG. There seemed to be a trend of increasing seropositivity with age in all three groups, however only parovirus B19 reached statistical significance. The incidence of all three infections were higher among the Malays, Indians and other races compared to the Chinese. CONCLUSION: CMV is endemic in our population and hence the most common infection. Toxoplasmosis and parvovirus is relatively low in our population but this implies that a large proportion of our antenatal women are still susceptible to these infections. Prevention of congenital CMV, toxoplasmosis and parvovirus infection is mainly by educating the antenatal population. PMID- 11063179 TI - A review of inpatients with adverse drug reactions to allopurinol. AB - Allopurinol is still an effective uric-acid lowering drug since its introduction in 1963. However it has been frequently incriminated for severe adverse drug reactions. From our retrospective review of 13 inpatients with allopurinol adverse reactions seen over 3 years, fever and rash were the commonest presenting symptoms, occurring several weeks after initiation of the drug. Other associated abnormalities included leukocytosis (62% of patients), eosinophilia (54%), renal impairment (54%) and liver dysfunction (69%). Although 12 patients (92%) met the criteria for allopurinol hypersensitivity syndrome, there was no mortality recorded. The indications for initiating allopurinol therapy were frequently unclear. In view of the severe adverse reactions experienced with allopurinol, we propose that it should only be prescribed when truly indicated. PMID- 11063180 TI - Stereotactic brain biopsies in AIDS patients--early local experience. AB - AIM: To assess the usefulness of stereotactic brain biopsies in AIDS patients with cerebral lesions in Singapore. METHODS: A total of 10 patients with AIDS and cerebral lesions underwent stereotactic brain biopsies in the Department of Neurosurgery, Tan Tock Seng Hospital (TTSH) between September 1997 and September 1998. The patients were referred from the Communicable Diseases Centre (CDC), TTSH. These patients either failed a trial of therapy for toxoplasmosis encephalitis (TE) or had CT/MRI scans which did not suggest TE. Four were CT guided and six were MRI-guided stereotactic biopsies. The Radionics Cosman-Robert Wells (CRW) stereotactic apparatus was used for all cases. RESULTS: The male to female ratio was 9:1. Histological diagnosis from biopsy was lymphoma (5), metastatic adenocarcinoma (1), TE (1), abscess (1), encephalitis (1) and granulomatous tissue (1-presumed tuberculosis). CONCLUSION: The early experience is that stereotactic brain biopsy is useful in patients with AIDS and cerebral lesions. The etiology is confirmed in the majority of cases and impacts on management decisions and prognostication. PMID- 11063181 TI - Family influence on current smoking habits among secondary school children in Kota Bharu, Kelantan. AB - OBJECTIVES: To measure the prevalence of cigarette smoking among male secondary school children and assess their family influence especially that of their fathers' smoking habits on their current smoking habits. METHODOLOGY: A cross sectional study was carried out in Kota Bharu, Kelantan in April 1997 where 460 male form four students, aged 15-16 years were randomly selected from six secondary schools. Data on smoking habits, sociodemographic profile and family characteristics particularly parents and siblings' smoking habits, perceived parental supervision and communication were collected through self-administered questionnaires. RESULTS: The prevalence of cigarette smoking among male secondary school children was 33.2%. Crude analysis shows family factors, fathers' and siblings' smoking habits, and lack of parental supervision were significantly associated with the students' current smoking habit. Among students who smoked compared to non-smokers, father's smoking habit gives a crude Odds Ratio = 1.8, 95% C.I. 1.08 - 3.16. Further analysis shows that the effect of their father's smoking habit on the student's current smoking habit is still significant after controlling for other familial and non-familial factors including parental supervision, academic performance, reported influence of cigarette advertisement, having friends who smoked and the student's poor knowledge of the ill-effects of smoking and other factors (Odds Ratio = 1.9, 95% C.I 1.05 - 3.32). In conclusion, family factors especially the father's smoking habit is an important factor that influences a student's current smoking habit and the presence of negative role models within the home need to be seriously considered in any cigarette smoking prevention programs among secondary school adolescents. PMID- 11063182 TI - Hypotension in acute myocardial infarction patients given streptokinase. AB - OBJECTIVE: This is a prospective cohort study done over a period of one year to look at hypotension that developed in the local Acute Myocardial Infarction (AMI) patients given Streptokinase (SK). METHOD: Suitable patients with AMI (those with ischaemic chest pain most severe within the last 8 hours, ST-segment elevation and no contraindications) were selected for thrombolysis with SK given as the standard dose of 1.5 mega-units diluted in 100 mls of normal saline and infused over 60 minutes. (Group A). The AMI patients who did not receive SK (Group B), were analysed separately and acted as "controls", as it was not possible to withhold thrombolytic therapy in a group of patients in a completely randomised fashion. The pulse, non-invasive blood pressure and electrocardiogram were monitored and recorded. RESULTS: Of 120 patients analysed, 70 received SK (Group A) and 50 (Group B) did not due to a variety of reasons. There was no statistically significant difference in the sex, age and body weight distribution as well as the initial mean arterial blood pressure (MAP) in the two groups. The MAP showed a statistically significant decrease at 15 minutes (105.6 to 81.4 mmHg, 95%CI: 13.965, 28.178) and 30 minutes (105.6 to 89.6 mmHg, 95%CI: 10.929, 19.814) after the commencement of SK in Group A patients. When analysed separately, the decrease in MAP was also statistically significant at 15 minutes (95%CI: 4.263, 22.014) for those with anterior AMI and both at 15 (95%CI: 19.112, 41.299) and 30 minutes (95%CI: 1.191, 28.716) for those with inferior AMI. There was no statistically significant decrease noted in Group B patients and the door to-needle time for Group A patients was 37.2+/-6.0 minutes. The SK infusion time for Group A patients who developed hypotension was prolonged to 95.3+/-14.1 minutes. CONCLUSION: Hypotension was more commonly noted in the AMI patients given SK. The MAP tend to decrease in the first 30 minutes after commencing the SK infusion. It is thus possible to conclude that the hypotension was at least partly due to SK and is probably a rate-related phenomenon. PMID- 11063183 TI - Staphylococcus lugdunensis: report of first case of skin and soft tissue infection in Singapore. AB - We report the first case of skin and soft tissue infection due to Staphylococcus lugdunensis in Singapore. This is a coagulase negative Staphylococcus species known to cause a wide variety of more serious infections--brain abscess, sepsis, chronic osteomyelitis and infective endocarditis. PMID- 11063184 TI - Cutaneous leishmaniasis: a report of two cases seen at a tertiary dermatological centre in Singapore. AB - Cutaneous leishmaniasis (CL) is not common in South-East Asia and often presents as a granulomatous plaque on the exposed areas, with a high index of suspicion required for diagnosis. Two such cases were seen at the National Skin Centre recently, and both were Gurkha men with a history of travel to Belize. They were treated with intravenous sodium stibogluconate with success. A discussion on CL and its management follows. PMID- 11063185 TI - Rhabdomyolysis and acute renal failure complicating detergent ingestion. AB - Rhabdomyolysis should be considered in the aetiology of all patients with unexplained acute renal failure (ARF). Early recognition provides the opportunity to initiate therapy aimed at preventing or limiting nephrotoxicity from the released heme pigment, myoglobin. We report an adult patient who developed ARF following the ingestion of a large amount of household detergent which, as far as we are aware, has not been previously described. The report illustrates the importance of measuring muscle enzyme levels and urinary myoglobin to confirm the possibility of rhabdomyolysis in any unusual presentation of ARF. PMID- 11063186 TI - Sphenochoanal polyps in Singapore: diagnosis and current management. AB - Sphenochoanal polyp is a rare form of choanal polyp. If unrecognised, they can be mistaken for an antrochoanal polyp. This will result in unnecessary exploration of the maxillary sinus, and a failure to remove the sphenoidal component of the sphenochoanal polyp. Adequate preoperative evaluation with computed tomography or magnetic resonance is mandatory to ascertain the correct diagnosis, and to facilitate the planning of the appropriate surgical procedure. We present two patients with sphenochoanal polyp and a review of the literature. PMID- 11063187 TI - Fecal incontinence: hope for an underdiagnosed condition. AB - Fecal incontinence is often suffered in silence leading one to become a social recluse. This has led to the belief that the problem does not exist and therefore underdiagnosed. In the last decade, much has been learnt about the understanding of continence and defecation. Quantification of appropriate physiological parameters associated with fecal incontinence have allowed the patients to be assessed using a logical algorithm. More importantly, parallel developments in management techniques of fecal incontinence now allow the categorised patients to be managed logically. The assessment is thus translated into appropriate management plans which range from simple nonoperative medical and dietary manipulation to pelvic floor retraining, sphincter augmentation and finally sphincter reconstruction. PMID- 11063189 TI - Common fallacies in the management of Grave's disease. PMID- 11063188 TI - Clinics in diagnostic imaging (48). Cystic liver metastases. AB - A 51-year-old man presented with an epigastric mass. Ultrasonography showed multiple cystic liver masses due to metastases from transitional cell carcinoma. Causes of cystic liver lesions include simple cyst, polyeystic liver disease, abscess, choledochal cyst, biliary cysadenoma and cystadenocarcinoma, and primary liver tumour. The imaging features of various types of cystic liver lesions are reviewed. PMID- 11063190 TI - Predictive factors of post-discharge mortality in the hospitalised elderly. AB - AIM: To study the death rate of noninstitutionalised elderly after discharge from hospital, describe the causes of death and identify predictive factors of mortality. METHODS: Vital status and cause of death of patients was ascertained by linkage to the death registry I year post discharge. Age, sex, race, marital status, housing and class of ward (to reflect socioeconomic status), presence of carer, Elderly Cognitive Assessment Questionnaire (ECAQ) score, Barthel score, presence of depression, number of chronic illnesses and length of stay in the hospital were studied as potential predictive factors for mortality using Cox proportional hazard regression models. RESULTS: Death occurred in 38 out of 113 patients. The commonest causes of death were malignancy (18%) and cerebrovascular disease (18%). Barthel score and length of stay were independent significant predictive factors of mortality. Elderly with low ECAQ score, with carers and from C class wards also had higher mortality although these results were not statistically significant. CONCLUSION: Post-discharge mortality is high in the elderly. We recommend that further studies be done to determine if amelioration of these predictive factors would lead to decreased mortality or improvement of quality of life. PMID- 11063191 TI - Ambulatory blood pressure monitoring in passive smoking and atropine--response in healthy volunteers. AB - AIM: The purpose of this study is to assess the effects of intravenous atropine on blood pressure (BP), systolic BP (SBP) and diastolic BP (DBP), after exposure of healthy volunteers to environmental cigarette smoking. METHODOLOGY: Seventeen non-smoking healthy volunteers (11 men and six women), aged from 20 years to 50 years (mean age: 31.3 +/- 8.7), were studied. These subjects received atropine under two conditions: first, in a 60 cubic metres of indoor space unpolluted by cigarette smoke; and second, in the same environment but polluted by 35 ppm carbon monoxide concentration reached by cigarette combustion. BP of each subject was recorded every 15 minutes for two hours by using an Ambulatory Blood Pressure Recorder BR 102. RESULTS: Mean baseline BPs in a smoke-free environment and in a smoking environment were respectively 120 +/- 18 mmHg and 120 +/- 25 mmHg for systolic values, and 81 +/- 6 mmHg and 84 +/- 9 mmHg for diastolic values (P > 0.05). After administration of atropine, mean BPs were respectively 126 (+/- 24)/80(+/- 7) mmHg in a smoke-free environment, and 131 (+/- 14)/ 90(+/- 2) mmHg after exposure to passive smoking (P > 0.05, no statistical significance). CONCLUSION: Our observations showed that BP varied but without statistically significant changes after acute exposure to passive smoking. However, it is known that any increase in BP leads to higher cardiovascular risks. PMID- 11063192 TI - The operative treatment of closed tibial fractures. AB - The tibia is a subcutaneous bone. Operative fixation of tibial fractures is a demanding undertaking. Thirty-five patients with 36 tibial fractures were admitted to our institution between May 1995 and April 1996. The patients were predominantly male (male to female ratio of 4) and the average age of the patients was 31.4 years (range 14 to 67 years). Ten fractures were located in the proximal third, 18 in the middle third and 8 in the distal third. The indications for operation included displaced intra-articular fragments, failed conservative treatment, compartment syndrome, multiple fractures and unstable fracture configuration. Operative procedures included plating in 29 cases and nailing in 7 cases. These patients were reviewed retrospectively and assessed for complications and radiological and functional outcome. The overall results were satisfactory in 88.9% and poor in 11.1%. The complications were reviewed and various factors affecting the incidences analysed. Three deep infections occurred. All were found after discharge from inpatient care. A prolonged interval between admission and surgery as well as high energy of impact are thought to be the main contributing factors. PMID- 11063193 TI - Routine screening for Chlamydia trachomatis in subfertile women--is it time to start? AB - Genital Chlamydia trachomatis infection has long been recognised as the major cause of pelvic disease and subsequently infertility. The diagnosis of this infection has traditionally relied on tissue culture. The availability of DNA amplification methods like ligase chain reaction promises faster and more sensitive results. This study was conducted to evaluate the prevalence of chlamydial infection in a subfertile population subgroup. AIM: A case control longitudinal study of 100 subfertile women in a tertiary teaching hospital were analyzed for the prevalence of genital Chlamydia trachomatis infection using ligase chain reaction test kit. RESULTS: A prevalence rate of 8% was detected, the majority being 25 years old or less (33.3%), p = 0.007. All patients gave no prior history of abnormal PAP smears, hospitalisation for pelvic inflammatory disease or abnormal vaginal discharge at the time of investigation. CONCLUSION: Our infertile group of patients has a relatively high incidence of silent genital Chlamydia trachomatis infection. This being highest in the below 25 years old age group. This finding indicates that screening for chlamydia may be necessary for the subfertile couple presenting to clinic. This is especially so if the patient is of the younger age group. PMID- 11063194 TI - A review of offenders remanded in a State Psychiatric Hospital. AB - Patients remanded over a two-year period to Woodbridge Hospital by Court Order were studied retrospectively. Schizophrenia was the most common diagnosis, theft and robbery the most prevalent offences. Males greatly outnumbered females. There were important gender differences, with males tending to commit sexual offences and females, theft and mischief. Males were less likely to be acquainted with their victims but those who caused hurt were more likely to know their victims. Outrage of modesty and theft were more likely to be committed against strangers. The reconviction rate was 26%, with repeat offenders more likely to commit sexual offences and theft. Patients who had previous psychiatric hospitalisation were more likely to be attending follow-up prior to and after release from remand and were more likely to have schizophrenia. Those assessed to be fit to plead were either fined or given jail sentences. Unsoundness of mind and unfitness to plead were associated with further remand in this hospital. PMID- 11063195 TI - Clinical characteristics and treatment outcome of 218 patients with non-Hodgkin's lymphoma in a Singaporean institution. AB - A retrospective analysis of 218 patients with non-Hodgkin's lymphoma (NHL) seen at a single institution in Singapore over a ten-year period was conducted. Twenty percent, 56% and 24% of patients had low-, intermediate- and high-grade disease respectively using the Working Formulation, and 25% of patients immunophenotyped had T-cell NHL. Forty-nine percent had primary extranodal disease, with the commonest sites of involvement being the gastrointestinal tract, nasal cavity, and tonsils. 86% and 73% respectively of patients with intermediate and high grade disease received combination chemotherapy as first line treatment, with CHOP being the most commonly used regime. Seventy-four percent of patients with low grade lymphoma received first line chemotherapy, 5% each was treated with radiotherapy or surgery alone, and 21% was treated symptomatically. Patients with low grade B-cell lymphoma had a 5-year survival of 80% and 10-year survival of 42%. One-year survival for intermediate and high grade B-cell lymphoma was 76% and 42%, while 2-year survival was 67% and 42% respectively. 1-, 2-, and 3-year survival for patients with T-cell lymphoma was 67%, 46% and 37% respectively. The difference in survival between low-, intermediate- and high-grade B-cell lymphoma was statistically significant (p = 0.0018 using the log rank test), but that between B- and T-cell lymphoma was not. Using the Cox regression model, International prognostic index, grade and extranodal disease were found to be statistically significant predictors of survival (p = 0.0001, p = 0.0157, p = 0.0343) respectively. PMID- 11063196 TI - Cutaneous Rosai-Dorfman disease--a pathologic review of 2 cases. AB - Rosai-Dorfman disease, also known as sinus histiocytosis with massive lymphadenopathy, is a rare but distinct clinicopathologic entity characterised histologically by a benign s histiocytic proliferation. Isolated involvement of extranodal sites without concomitant nodal disease is rare. We describe the pathological features of 2 cases of Rosai-Dorfman disease that were clinically confined to the skin. In both male adult Chinese patients, proliferation of histiocytes was accompanied by S-100 protein expression demonstrated immunohistochemically within the histiocytes. The pathology of Rosai-Dorfman disease and its microscopic differential diagnoses are discussed. PMID- 11063197 TI - Complete hydatidiform mole and surviving coexistent twin--a case report. AB - Hydatidiform mole with a coexistent fetus is a rare occurrence with an incidence of I per 22,000-100,000 pregnancies. It is associated with persistent gestational trophoblastic tumour. Hence an early and correct diagnosis is imperative to plan subsequent management of such patients. We report a case of a primigravida who presented with vaginal bleeding at early second trimester. Expectant management was carried out for her pregnancy which finally ended in an abortion. The pathology, clinical findings and current management of this rare entity is discussed. PMID- 11063198 TI - Primary leiomyoma of the liver. AB - This case report describes a primary hepatic leiomyoma presenting as a mass lesion detected on ultrasonography of the abdomen in an asymptomatic hepatitis B carrier on routine surveillance. Primary leiomyomata of the liver are rare occurrences, with only 9 cases reported in the literature. The presenting features of primary hepatic leiomyomata and diagnostic approach towards such lesions are discussed. The significance of such tumours in the immunocompromised is also mentioned. PMID- 11063199 TI - The value of MRI in the diagnosis and management of neurocysticercosis. AB - The clinical manifestation of neurocysticercosis is quite protean and variable making it the 'great imitator' of almost any neurological disorder. In the last decade, developments in diagnostic imaging and effective anticysticercus drug therapy have changed the outlook of the disease. Two cases investigated with CT scan and MRI are reported here. One case was treated effectively with antihelminthic therapy. In both cases MRI was found to be much more sensitive than CT scan in picking up the multiple cystic lesions. In addition, 'protoscoleces' and 'differential ring enhancements' not apparent in CT scan were well shown in MRI hence enabling the demonstration of different activity stages. In the treated patient, repeat MRI showed degeneration of the cyst. These illustrate that MRI is superior to CT scan in both the diagnosis and management of the illness. PMID- 11063200 TI - Clinics in diagnostic imaging (47). Huntington's disease. AB - A 38-year-old man presented with progressive worsening of choreiform movements. Serum biochemistry analysis did not reveal any abnormality. Magnetic resonance imaging demonstrated symmetrical caudate nucleus atrophy and generalised cerebral strophy. Huntington's disease was diagnosed in view of the clinical presentation and the characteristic imaging findings. The clinical, pathological and imaging features of this disease process are discussed. PMID- 11063201 TI - Electrocardiographic case: diagnosis of acute myocardial infarction in the presence of left bundle branch block. AB - The electrocardiographic features associated with acute myocardial infarction (AMI) are often easily recognised. However, interpretation is made more difficult in the presence of confounding patterns such as a left bundle branch block (LBBB). This may result in missed cases which may otherwise have benefited from acute revascularisation therapy. Though not straightforward, the diagnosis of AMI in the presence of LBBB can be made with a reasonable amount of accuracy. We report a case of acute myocardial infarction with LBBB that was appropriately diagnosed and underwent acute revascularisation by angioplasty. A detailed knowledge of the typical electrocardiographic features associated with LBBB, especially the ST segment morphologies, is very important. This will greatly aid recognition of an evolving AMI and help us decide on the most appropriate therapy. PMID- 11063202 TI - "The pain of the mind is worse than the pain of the body". PMID- 11063203 TI - Attitudes and beliefs of Singapore health care professionals concerning HIV/AIDS. AB - AIM OF STUDY: This study examines the beliefs and attitudes of Singapore doctors, dentists, and nurses concerning HIV/AIDS and persons living with HIV/AIDS (PWAs). METHOD: A mail survey was done of all doctors and dentists in Singapore as well as a random sample of 1,500 nurses from the Singapore Nursing Board Register. RESULTS: The results showed that respondents held accurate beliefs concerning transmission of HIV via sex and needle sharing but a significant proportion also expressed belief in transmission via everyday social contact. Respondents were aware of universal precautions when treating persons with HIV/AIDS but tended to be overly cautious in low/no risk situations. A substantial proportion of respondents indicated little or no knowledge or experience with AIDS-related conditions and the majority believed that most health care professionals are unprepared to care for PWAs. Further, there was evidence of substantial stigmatisation and fear of treating PWAs, both of which were significantly and negatively correlated with accuracy of beliefs about HIV transmission and universal precautions. CONCLUSION: These results point to important misconceptions about HIV/AIDS held by Singapore health care professionals as well as stigma towards and fear of treating PWAs. These are areas that need to be addressed through better professional education concerning HIV/ AIDS. This education needs to address both the factual misconceptions about HIV/AIDS as well as the stigma associated with this disease and the fears that health care professionals have of treating PWAs. PMID- 11063204 TI - Wegener's granulomatosis in the elderly. AB - Wegener's granulomatosis classically involves the upper respiratory tract, lungs and kidneys. Rarely, it also affects the skin and heart. Cardiac involvement is uncommon in Wegener's granulomatosis and myocardial infarction is seldom highlighted. It can be a difficult diagnosis to make in the elderly who often have multiple co-existing illnesses. We present a case of a 75-year-old Chinese woman with interesting cardiac and dermatological manifestations of Wegener's granulomatosis. PMID- 11063205 TI - A pilot study on the role of intralesional triamcinolone acetonide in the treatment of pitted nails in children. AB - BACKGROUND: Pitted nails is a non-specific entity seen in children that is often associated with various underlying skin disorders such as atopic dermatitis, psoriasis, alopecia areata and lichen planus; and those without such associations are labelled as idiopathic pitted nails. Spontaneous resolution is common but may take several years. Treatment options so far have not been encouraging. This pilot study sought to determine the safety and efficacy of intralesional steroid in the treatment of pitted nails in children. METHOD: Children with pitted nails below the age of 12, seen between January 1994 and December 1997, were invited to participate in this study. Symmetrically affected fingernails were selected, topical anaesthetic cream applied and a single dose of triamcinolone acetonide was introduced intralesionally to the proximal nailfold while the contralateral nail acted as control. The degree of pitting of nail surface was documented at months 0, 2 and 4. RESULTS: Four children were enrolled, and their ages ranged from 4 to 9 years. The degree of pitting was reduced to a mean of 15% of the nail surface in the second month, and 42% in the fourth month. The procedure was fairly well tolerated and no adverse effects were noted. CONCLUSION: Intralesional steroid is a safe and acceptable method of treating pitted nails in children with a maximum effect seen in 2 months. In order to sustain a continuous therapeutic effect, a bimonthly treatment is suggested. PMID- 11063206 TI - The use of abciximab in coronary angioplasty--an Asian centre's experience. AB - BACKGROUND: Abciximab, a monoclonal antibody to platelet glycoprotein IIb/IIIa receptor, has been shown to be effective in reducing ischemic complications after coronary angioplasty in recent trials. However, little is known about its efficacy and safety when used in Asian patients. METHODOLOGY: Based on our abciximab registry, we performed a retrospective analysis of 115 Asian patients who received the antiplatelet agent while undergoing percutaneous coronary intervention in our centre. They constituted 18.4% of the total number of patients undergoing percutaneous revascularisation during the corresponding period. The majority of the patients were males (84%). The mean age of the cohort was 54 and the mean weight was 70 kg. The ethnic composition of the study population was: Chinese 54%, Indians 21%, Malays 19% and Others 6%. All patients received aspirin 100 mg and weight-adjusted heparin before the procedure. Abciximab may be administered on a preplanned basis prior to the procedure or be given as a 'bailout' strategy. RESULTS: There was a high clinical success rate of 95.8% and low incidence of ischemic complications when abciximab was given during coronary angioplasty. There were 0% Q myocardial infarction, 3.3% non-Q myocardial infarction and 0.8% death in our series. Bleeding complications were uncommon at 7.6%, predominantly involving the groin and gingiva. Thrombocytopenia occurred in 5.8% of patients. Abciximab was noted to increase the procedural activated clotting time (ACT) by 38 seconds when given concomitantly with heparin. The mean maximal procedural ACT achieved was 323 +/- 51 seconds. CONCLUSIONS: Abciximab may be used safely and efficaciously in Asian patients undergoing coronary angioplasty. The drug confers protection against ischemic complications during the procedure whether it is administered electively or as a 'bailout'. There is however, a need to redefine the heparin regime for our patients, given the high ACT obtained when abciximab is administered. PMID- 11063207 TI - A man with osteoblastic metastasis and hypocalcaemia. AB - We report a case of an 80-year-old man with osteoblastic metastases from advanced carcinoma of the prostate presenting with a grand mal seizure resulting from severe hypocalcaemia. He had low serum phosphate and ionised calcium levels, elevated serum skeletal alkaline phosphatase and intact parathormone levels. 99mTc radioisotope bone scan revealed a "super bone scan" suggestive of osteomalacia. The serum 1, 25-dihydroxycholecalciferol level was unexpectedly elevated. The biochemical abnormalities persisted despite high dose calcium replacement, but improved with supraphysiological doses of 1,25 (OH)2 vitamin D3 (Rocaltrol) therapy. We hypothesise that the hypocalcaemia in this patient was due to vitamin D resistance secondary to a humoral factor secreted by the tumour. PMID- 11063208 TI - Migrating oesophageal foreign body--an unusual case. AB - Ingested foreign bodies which migrate extraluminally are rare occurrences. If untreated, they may result in life threatening complications. Exploration of the neck via an external approach to remove the foreign body is the recommended treatment. The CT Scan utilising fine cuts is invaluable in localising the foreign body for exploration. The case of a patient with a metallic foreign body in the oesophagus which migrated extraluminally is presented. Hemithyroidectomy was required to gain access for removal of the foreign body. A discussion on the management of such a case follows. PMID- 11063209 TI - Cryptococcal meningitis resulting in irreversible visual impairment in AIDS patients--a report of two cases. AB - Cryptococcus neoformans is the leading cause of meningitis in patients with Acquired Immune Deficiency Syndrome (AIDS) and is associated with high mortality rate. Presenting symptoms include fever, nausea and vomiting, altered mentation, headache and meningismus. Cryptococcal meningitis is not infrequently complicated by raised intracranial pressure and visual sequelae (sometimes by blindness). In patients who survive the infection, the most debilitating outcome appears to be visual impairment or blindness. Management of impending visual complication combines medical and surgical treatment modalities. We report two cases of cryptococcal meningitis associated with visual impairment. PMID- 11063210 TI - Boerhaave's syndrome (ruptured oesophagus) in a case of rabies. AB - A 46-year-old Caucasian man was admitted with a history of dog-bite in a foreign country six months previously. He presented with multisystem complaints, died suddenly soon after admission and the only significant finding at post-mortem was a ruptured oesophagus. Immunological tests confirmed rabies. PMID- 11063211 TI - High dose chemotherapy in breast cancer--a thin red line between hype and hope. PMID- 11063212 TI - Clinics in diagnostic imaging (46). Renal cell carcinoma in acquired cystic kidney disease. AB - Acquired cystic disease of the kidney and renal cell carcinoma are associated with chronic renal failure. In recent years, there has been increased incidental detection of renal tumours through the liberal use of ultrasonography (US) and computed tomography (CT). A 40-year-old man suffering from chronic renal failure and who was being treated with haemodialysis for six years, was found to have a complex cystic lesion on US and CT. Nephrectomy was performed and the lesion was confirmed to be renal cell carcinoma associated with acquired cystic kidney disease. The current role of imaging in the management of affected patients are discussed. PMID- 11063213 TI - What you need to know: diving medicine and the role of the family physician. PMID- 11063214 TI - Medical responsibility in a hospital practice. AB - The complex legal relationships between hospitals, doctors and paramedical staff lead to issues which the court will find difficult to resolve. However, certain trends have emerged in modern medicine: 1) There is a need to provide competent care based on a national standard. 2) Competent care is no longer predicated on 'locality rules'. The state has to intervene with statutes and regulations to ensure that a 'standard' of practice is established in hospitals. 3) The hospital has both a vicarious as well as an inherent duty of care (corporate obligation) to its patients. 4) The statutory regulations result in doctors being involved directly in setting of standards. This brings a separate liability upon the doctors independent of their professional liability. 5) There is a demand not only for establishing initial standards of care, but for continuous monitoring of these standards and proactive measures to ensure that they are updated. PMID- 11063215 TI - DNA methylation and gastrointestinal malignancies: functional consequences and clinical implications. AB - Evidence now suggests that the transcriptional silencing of selected genes by DNA methylation plays a crucial role in the development and progression of human gastrointestinal malignancies. To date, genes involved in the regulation of the cell cycle, DNA repair, angiogenesis, and apoptosis have all been shown to be inactivated by the hypermethylation of their respective 5' CpG islands. The specific causes and effects of the changes in DNA methylation that occur in cancer remain unknown, however. Nevertheless, recently developed techniques for detecting changes in DNA methylation have dramatically increased the amount of information available on the patterns of methylation that occur as cancers progress. One key process involved in aberrant methylation is related to aging, and because it affects a large number of CpG islands, age-related methylation may be a primary cause of the increased incidence of cancer seen among older individuals. Other patterns of methylation are cancer-specific and are detected only in a subset of tumors exhibiting the CpG island methylator phenotype (CIMP). In this regard, the majority of sporadic colorectal cancers, exhibiting microsatellite instability (MSI) appear to be associated with CIMP, which leads to aberrant methylation of human MutL homologue (hMLH1) and the loss of its expression. We anticipate that as the various components of the molecular machinery involved in aberrant DNA methylation become better understood, they will prove to be useful targets, serving as the basis for the development of new methods for the diagnosis and treatment of gastrointestinal malignancies. PMID- 11063216 TI - Effects of Helicobacter pylori infection on Zollinger-Ellison syndrome. AB - Both Zollinger-Ellison syndrome (ZES) and Helicobacter pylori infection are major etiologic factors for peptic ulcer. The aim of this study was to investigate the effect of H. pylori infection on ZES with special reference to acid secretion. Sixteen patients with ZES were selected (median age, 59 years; range, 39-66 years; M/F, 9/7), and H. pylori status, ulcer location, gastric acid secretion, serum pepsinogen (PG) I and II concentrations, and PG I/II ratio were determined. The seroprevalence of H. pylori infection was 50%, whereas active H. pylori infection was seen in only 25% of the patients. Thirteen patients had duodenal ulcer (DU), 1 had gastric ulcer (GU), and 2 had both GU and DU. DU was seen in both H. pylori-positive and H. pylori-negative patients, whereas GU was found only in H. pylori-positive patients. Both basal and maximal acid outputs were significantly lower in H. pylori-positive patients than in H. pylori-negative patients (P< 0.05). Moreover, both serum PG I and the PG I/II ratio were significantly lower in H. pylori-positive patients than in H. pylori-negative patients. These results indicate that ZES is an independent risk factor for DU, but H. pylori infection may play some role in the development of GU in ZES. In patients with ZES, H. pylori infection may reduce both hypersecretion from parietal cells and PG I secretion from chief cells, and hyperacidity of the stomach in ZES may have eradicated H. pylori in some patients. PMID- 11063217 TI - Effect of a histamine H1 receptor antagonist on gastric endocrine cell proliferation induced by chronic acid suppression in rats. AB - The effect of histamine on gastrin cells and enterochromaffin-like cells has not yet been clarified. We investigated the influence of pyrilamine (a histamine H1 receptor antagonist) on serum gastrin level, gastrin cells, and enterochromaffin like cells in rats with or without 4 weeks of famotidine treatment. The rats were divided into six groups: a control group, two pyrilamine groups (2mg/kg, or 20mg/kg, p.o.), a famotidine group (20mg/kg twice/daily i.m.), and two pyrilamine + famotidine groups. The serum gastrin concentration was determined, and gastrin cells and enterochromaffin-like cells were identified by the labeled streptavidin biotin complex method and counted. Hypergastrinemia, gastrin cell hyperplasia, and enterochromaffin-like cell hyperplasia were found after 4 weeks of famotidine treatment. Four weeks of treatment with pyrilamine alone did not affect the gastrin level, gastrin cells, or enterochromaffin-like cells in the rat stomach. When combined with famotidine, pyrilamine enhanced famotidine-induced hypergastrinemia, but it did not affect gastrin cell hyperplasia, and it significantly inhibited enterochromaffin-like cell hyperplasia. These results suggest that gastrin secretion and enterochromaffin-like cell proliferation may be regulated by histamine via the H1 receptor during acid suppression. PMID- 11063218 TI - Vital immunostaining of human gastric and colorectal cancers grafted into nude mice: a preclinical assessment of a potential adjunct to videoendoscopy. AB - Videoendoscopy has not significantly advanced diagnostic accuracy beyond that attainable by conventional fiberscopy, with respect to microcarcinomas of the digestive tract. We suspected that after the labeling of these lesions with an agent detectable by videoendoscope, digital processing of the images could facilitate endoscopic diagnosis of microcarcinomas. We have developed a novel antibody labeled with an indocyanine green (ICG) derivative that is evident by videoendoscope. However, the binding of such an exogenous antibody in vivo to tumor surfaces has not been described. In this preliminary study, after transplanting human gastric cancer or colorectal cancer into nude mice, we successfully bound the tumors in vivo with an anti-MUC1 mucin antibody, as subsequently confirmed by the performing of immunohistochemistry with a secondary antibody. The antibody labeled with an ICG derivative may therefore be clinically useful in detecting gastrointestinal microcarcinoma by videoendoscopy. PMID- 11063219 TI - Progressive and sufficient decrease of hepatitis B core antibody can predict the disappearance of hepatitis B virus DNA in Japanese patients with hepatitis B surface antigen clearance. AB - The aim of this study was to elucidate the relationships among serum levels of hepatitis B virus (HBV) DNA, periods after hepatitis B surface (HBs) antigen clearance, and the titer of hepatitis B core (HBc) antibody in 200-fold diluted serum. Twelve patients who had clearance of HBs antigen from serum were studied. Five patients had not received any treatment (group A), and seven had received prednisolone withdrawal therapy. The patients in groups A and B were followed up for 86 months and 108 months (median), respectively. Serum HBV was measured by the nested polymerase chain reaction method. In both groups, serum HBV tended to become gradually undetectable after HBs antigen clearance. The positive rate of HBV in the sera 5 years or more after HBs antigen clearance was significantly lower than that in the sera at less than 5 years, both in group A (P = 0.004) and group B (P = 0.010). In both groups, the titer of HBs tended to decline every year after HBs antigen clearance. HBV was still detectable in the sera of some patients for a long period of time after they showed seroconversion to HBs antibody. The results suggest that detection of HBV was difficult in sera with an HBc titer of 30% or lower and at more than 5 years after HBs antigen clearance in both groups. It is important to note that HBV DNA rarely exists in the serum, even when HBs antigen and HBc are both negative. PMID- 11063220 TI - The tight junction of pancreatic exocrine cells is a morphometrically dynamic structure altered by intraductal hypertension. AB - The tight junction of pancreatic exocrine cells is thought to regulate paracellular permeability, and is a possible reflux route of pancreatic juice into the blood flow. Morphological changes in the tight junction of canine pancreatic acinar cells following intraductal hypertension and secretin stimulation were morphometrically analyzed to obtain evidence of the control of the paracellular reflux. Pancreatic tissues obtained from 25 dogs after intraductal hypertension, 3 dogs after secretin stimulation, and 5 control dogs were studied. Intraductal pressure was either 20 cmH2O, 30 cmH2O, or 40 cmH2O. Freeze fracture replicas of these pancreatic tissues were observed by electron microscopy. Tight junctions were classified into six morphometric types. Reticular type, parallel type, and mixed type comprised the common types predominantly found in all groups, and three special types were found, infrequently, only after intraductal hypertension. The percentages of the common types were significantly different between the groups. The areas of the tight junctions, and other morphometric parameters, were significantly less after 20 cmH2O intraductal hypertension and secretin stimulation than in the controls. However, these findings after 30 cmH2O or 40 cmH2O intraductal hypertension did not differ from those in the controls. The areas of the three special types of tight junctions were larger than those of the common types. These results suggest that the tight junction of pancreatic exocrine cells is a morphologically dynamic structure that is altered by the extent of intraductal hypertension, and support the hypothesis that paracellular permeability is the mechanism of the reflux of pancreatic juice. PMID- 11063221 TI - Microsatellite instability in gallbladder carcinoma: two independent genetic pathways of gallbladder carcinogenesis. AB - Although the genetic basis for gallbladder carcinogenesis has not been clarified, considerable evidence has shown that genetic alterations play an important role in the development and progression of human cancers. In this study, we analyzed 30 gallbladder carcinomas to investigate the role of genetic alterations in their tumorigenesis, and to study correlations with their clinicopathological features. Tissue samples were obtained from 30 patients with gallbladder carcinoma (11 men and 19 women; mean age, 62 years; age range, 38-80 years). Genomic DNAs were extracted from fresh tumor tissue. We examined loss of heterozygosity (LOH) in the p53, APC, DCC, RB, and NM23-H1 gene regions by polymerase chain reaction (PCR)-LOH assay using an automated fluorescent DNA sequencer employing four microsatellite markers (p53, APC, DCC, NM23-H1). Five additional microsatellite markers were used for the determination of microsatellite instability (MSI). LOH was found at p53 in 9 of 15 informative cases (60%), at DCC in 10 of 22 (45%), at APC in 5 of 15 (33%), at RB in 1 of 8 (13%), and at NM23-H1 in 1 of 15 (7%). MSI was observed in 5 of 30 cases (17%) in at least one chromosomal loci of these nine microsatellite markers. None of the patients with MSI-positive tumors showed lymph node metastasis, and there was an inverse correlation between MSI and the presence of LOH in gallbladder carcinoma. These results suggest that there are two independent genetic pathways in gallbladder carcinogenesis; that is, an MSI pathway and an LOH pathway. PMID- 11063222 TI - Massive gastrointestinal bleeding from jejunal varices. AB - We report a patient with massive gastrointestinal bleeding from jejunal varices, confirmed by emergency laparotomy. A 54-year-old woman was admitted to Chonnam National University Hospital with a 5-day history of melena with hematochezia. Fifteen years previously, she had undergone cholecystectomy for gallstone. Seven years previously, she had undergone an operation because of possible common bile duct stone. The details of this operation were not known. Upper gastrointestinal endoscopy revealed no varices in the esophagus, stomach, and proximal duodenum. Colonoscopy demonstrated black-colored blood clots throughout the colon. A technetium-99m-labeled red blood cell (RBC) scan showed active proximal small bowel bleeding. Abdominal aortic angiography revealed mesenteric varices in the upper abdomen, but no active bleeding source was recognized. Because of the patient's continued massive gastrointestinal bleeding despite medical therapy, emergency laparotomy was performed. We found evidence of micronodular cirrhosis of the liver and an apparent Roux-en-Y anastomosis. There were numerous collateral variceal vessels in the jejunal limb with the liver and abdominal wall. Segmental resection of the involved jejunum and end-to-end anastomosis were perdilated formed. Histologic examination revealed submucosal veins with mucosal erosion. PMID- 11063223 TI - Specific mutation in exon 11 of c-kit proto-oncogene in a malignant gastrointestinal stromal tumor of the rectum. AB - Gastrointestinal stromal tumor (GIST) in the distal third of the rectum was detected in a 57-year-old man who underwent an abdominoperineal resection of the rectum. Because the tumor expressed CD34 and c-kit gene product, but did not express smooth muscle actin or S-100 protein, it was diagnosed as an uncommitted type of GIST. Moreover, a specific mutation in the sequence coding the juxtamembrane domain in exon 11 of the c-kit proto-oncogene was revealed by a polymerase chain reaction-single-strand conformation polymorphism method. One year after resection, the patient developed multiple liver metastases. It is suggested that a specific mutation in exon 11 of the c-kit proto-oncogene may have played an essential role in the development of the liver metastases. PMID- 11063224 TI - MRI facilitated a diagnosis of endometriosis of the rectum. AB - A 51-year-old pre-menopausal Japanese woman suffering from chronic lower abdominal pain was referred to our hospital. A barium enema showed a stenotic lesion in the recto-sigmoid region, and a pelvic computed axial tomography (CAT) scan revealed a thickened rectal wall. A colonoscopic examination showed the rectum to be constrictive, but the mucosa appeared to be intact. Magnetic resonance imaging (MRI) with T1 high-intensity revealed a cystic lesion in the thickened wall of the rectum, which led us to suspect possible bowel endometriosis. Part of the biopsy specimen showed endometrial epithelium within the interstitial layer of histologically normal mucosa; finally, endometriosis of the rectum was diagnosed. The patient became asymptomatic after the initiation of hormonal treatment and later experienced spontaneous menopause. MRI was effective for diagnosis and the patient did not undergo unnecessary laparotomy. Although bowel endometriosis is generally diagnosed by means of resected specimens, in our patient, diagnosis was made using MRI and biopsy, and hormonal therapy had an effective role as a bridge to menopause. PMID- 11063225 TI - Manifestation of cutaneous polyarteritis nodosa during interferon therapy for chronic hepatitis C associated with primary biliary cirrhosis. AB - Interferon alpha-2b was administered to a 50-year-old Japanese woman with chronic hepatitis C associated with primary biliary cirrhosis. Two months after the beginning of the interferon alpha-2b therapy a systemic nodular, erythematous rash developed. Histological analysis of the skin revealed typical features of necrotizing arteritis. Because there was no microhematuria, and no microaneurysms were detected on abdominal angiography, a diagnosis of cutaneous polyarteritis nodosa was made. A good outcome was achieved after interferon alpha-2b was discontinued and prednisolone was administered instead. The cutaneous polyarteritis nodosa in this patient is thus considered to have occurred as an adverse effect of interferon administration. To our knowledge, this is the first reported case of cutaneous polyarteritis nodosa which developed because of interferon therapy for chronic hepatitis C associated with primary biliary cirrhosis. PMID- 11063226 TI - What is the role of Helicobacter pylori infection in ZES-associated peptic ulcer? PMID- 11063227 TI - Can determination of HBcAB titer be a surrogate for HBV DNA assay? PMID- 11063228 TI - Microsatellite instability meets gallbladder cancer. PMID- 11063230 TI - Predicting language outcomes for young prelinguistic children with developmental delay. AB - The purpose of this study was to examine potential relationships between children's prelinguistic communication behaviors and subsequent (12 months later) expressive and receptive language outcomes. Participants included 25 toddlers with developmental delay and their mothers. The dyads were observed during natural interactions at 6-month intervals over a 12-month period for a total of 3 observation points (O1, O2, O3). Children's rate of nonverbal behavior that is often perceived as communication by adults was identified at O1 and O2. In the investigation, the children's intentional nonverbal communication acts all included coordinated attention between the communication referent and the adult. The other types of prelinguistic communication behavior, termed gestural indicating behavior and social interaction signals, were produced without coordinated attention to the adult. Receptive and expressive language test scores and spontaneous word productions were analyzed at O3 and served as outcome measures in regression analyses. Results indicated that rate of intentional nonverbal communication at O1 was a predictor of spontaneous word productions at O3. At O2, rate of intentional communication and rate of gestural indicating behavior predicted subsequent language outcomes as measured by the Sequenced Inventory of Communication Development-Revised. The results are consistent with previous findings for intentional nonverbal communication that includes coordinated attention, but additionally demonstrate that prelinguistic behavior lacking coordinated attention also bears a relationship to subsequent language outcome. Discussion of observed patterns focuses on child and adult factors that may motivate the transition from prelinguistic to early symbolic communication. PMID- 11063231 TI - Accuracy of metamemory after traumatic brain injury: predictions during verbal learning. AB - The primary intent of this study was to investigate the metamemory monitoring abilities of adult survivors of at least moderate traumatic brain injury (TBI) during a verbal-learning activity. Eighteen survivors and 18 non-injured control participants made judgment-of-learning (JOL) predictions of their recall ability immediately after studying 3 lists of noun-pairs or after a slight delay. A secondary intent of this study was to determine if verbal retrieval attempts would enhance predictive accuracy. One half of participants made retrieval attempts during the second and third list-learning task, and the other half made retrieval attempts during the third list-learning task only. Measures of the correlation between JOL predictions and recall accuracy revealed that survivors were as accurate as controls when making delayed predictions and were less accurate when making immediate predictions. This occurred regardless of retrieval attempts. Absolute measures that compared mean JOL ratings to overall recall revealed that the survivor group was well-calibrated when making delayed JOL predictions but overestimated when making immediate JOL predictions. The non injured control group underestimated when making both types of predictions. However, within-group variability was high. These findings are compared to those from studies that investigated metamemory beliefs in which survivors' ratings were compared to family-member ratings. Clinical implications for basing executive decisions about compensatory strategies on delayed and immediate predictions of future recall are discussed. Additionally, a rationale is provided for the use of both relative and absolute measures of predictive accuracy in metamemory studies involving neurological clinical populations. PMID- 11063232 TI - Validity of a parent-report measure of vocabulary and grammar for Spanish speaking toddlers. AB - The validity of the Fundacion MacArthur Inventario del Desarrollo de Habilidades Comunicativas: Palabras y Enunciados (IDHC:PE) was examined with twenty 20- and nineteen 28-month-old, typically developing, monolingual, Spanish-speaking children living in Mexico. One measure of vocabulary (number of words) and two measures of grammar (mean of the three longest utterances and grammatical complexity score) from the IDHC:PE were compared to behavioral measures of vocabulary (number of different words from a language sample and number of objects named in a confrontation naming task) and one behavioral measure of grammar (mean length of utterance from a language sample). Only vocabulary measures were assessed in the 20-month-olds because of floor effects on the grammar measures. Results indicated validity for assessing expressive vocabulary in 20-month-olds and expressive vocabulary and grammar in 28-month-olds. PMID- 11063233 TI - Directiveness in teachers' language input to toddlers and preschoolers in day care. AB - Five subtypes of directiveness were examined in the interactions of day care teachers with toddler and preschooler groups. The instructional context (book reading, play dough) yielded significant differences across all five subtypes of directiveness, indicating that these two activities elicited different types of teacher-child discourse. Book reading was characterized by significantly more behavior and response control and less conversation control in comparison with the play-dough activity. Correlations between teachers' directiveness and child language productivity indicated that behavior control and turn-taking control were associated with low levels of productivity, whereas conversation control was associated with the highest levels of productivity. The results of this study confirm that instructional context is an important mediator of teachers' directiveness and suggest that subtypes of directiveness have differential effects on child language output. PMID- 11063234 TI - The influence of argument-structure complexity on the use of auxiliary verbs by children with SLI. AB - The purpose of the present study was to investigate the influence of argument structure complexity on the omission of auxiliary be verbs in a group of children with specific language impairment (SLI). These children were compared to a group of younger, normally developing children matched for mean length of utterance (MLU) and a group of children matched for chronological age (CA). Using a story completion task, the children (N = 30) were required to produce sentences of varying length and argument-structure complexity. The results of the study indicated that the children with SLI omitted more auxiliary forms than either the MLU or CA controls. In addition, both the children with SLI and the MLU controls were more likely to omit the auxiliary forms when attempting sentences with greater argument-structure complexity. These results suggest that argument structure complexity may be a contributing factor to children's omissions of grammatical morphemes. PMID- 11063235 TI - Acquisition of irregular past tense by children with specific language impairment. AB - In this paper we add to what is known about the tense-marking limitations of children with specific language impairment (SLI) by exploring the acquisition of regular and irregular past tense, encompassing the age range of 2;6 to 8;9 (years;months) and comparing the performance of 21 children with SLI to that of 23 control children of the same age and 20 younger control children of equivalent mean length of utterance (MLU) at the outset. The analysis differentiated between the morphophonological component of past tense marking and the morphosyntactic component (finiteness). In the morphosyntactic component, the performance of the SLI group trails that of the two control groups over 3.5 years, whereas in the morphophonological component, the SLI group's performance is equivalent to that of the younger controls. Models of growth curves for regular past tense and irregular finiteness marking show the same pattern, with linear and quadratic components and the child's MLU at the outset as the only predictor. For morphophonological growth the picture changes, with an interaction of linear trend and MLU and the child's receptive vocabulary emerging as a predictor. The findings support a morphosyntactic model, such as the extended optional infinitive (EOI) model, with regard to the limitations in finiteness marking and for affected children. PMID- 11063236 TI - The relationship between event representation and linguistic skill in narratives of children and adolescents with Down syndrome. AB - Children and adolescents with Down syndrome present with greater difficulty in expressive language than nonverbal cognitive domains. As narratives involve an understanding of the relationship(s) between events and their verbal expression, this divergence has implications for understanding narrative abilities in persons with Down syndrome. In this project, we investigated the relationship between event representation and linguistic expression in narratives of children and adolescents with Down syndrome (n = 31) and groups of typically developing children matched for mental age (n = 31), syntax comprehension (n = 28), or expressive language (n = 27). A short wordless film, the Pear Story (Chafe, 1980), was viewed individually by each participant and then each participant retold the story to an adult who (presumably) had not seen the film. Findings suggest a disparate relationship between linguistic expression and event representation in narratives of children and adolescents with Down syndrome. Participants with Down syndrome produced narratives that were significantly longer and more complex than the expressive-language-matched-group, with no differences observed in event structure when compared to the MA-matched group. Comparatively, use of linguistic devices and cohesion were poorer in the children and adolescents with Down syndrome than in the MA-matched children, with no differences observed in comparison to children matched for expressive language. PMID- 11063237 TI - Practice and list effects on the synthetic sentence identification test in young and elderly listeners. AB - A high prevalence of central auditory processing disorders (CAPD) has been reported in the elderly based on scores for the Synthetic Sentence Identification Test (SSI). This study examined practice effects and list differences on the Auditec version of the SSI with an ipsilateral competing message in 30 young and 20 elderly participants. Results for the younger participants suggested that a minimum of one practice list should be used with the SSI, whereas results for the elderly participants suggested that a minimum of three practice lists should be used. The binomial model of A. R. Thornton and M. J. M. Raffin (1978) was suggested for use in determining the required number of lists needed to obtain reliable SSI performance. Significant list differences were found with the Auditec version of the SSI. It was recommended to use only Lists A, C, D, E, G, and I for consistent results across lists. Practice effects and list differences should be accounted for when using the SSI for the assessment of CAPD in elderly and young listeners. PMID- 11063238 TI - Tinnitus and cognitive interference: a stroop paradigm study. AB - The aim of this study was to investigate cognitive interference caused by tinnitus by means of a modified version of the Stroop color-word test. In a mixed design study, the performances of tinnitus patients (n = 23) and healthy controls with normal hearing (n = 23) were compared on three versions of the Stroop test: the original version, a modified version including physical-threat words, and a tinnitus version for which tinnitus words (descriptors of tinnitus; e.g., peep) were derived empirically. Matched control conditions (words) were included for all three versions, yielding a total of six screens that were presented on a computer. Participants in the control group were matched with the patients for age and gender. Main dependent measures were performance on the Stroop tests in terms of total time for completing each test. Also included were the Tinnitus Questionnaire (TQ), the Beck Depression Inventory (BDI), the state version of the Spielberger Trait State Anxiety Inventory (STAI-S), and a subtest from the Wechsler Adult Intelligence Scale measuring verbal ability. Results showed that tinnitus patients performed significantly slower on all six test conditions. The classical Stroop color-word interference was replicated in both groups. Also, an effect for physical-threat words was found for both groups. Our expected tinnitus word interference could not be established. Patients scored significantly higher than controls on the BDI and the STAI, but these measures did not correlate with the Stroop results. In conclusion, the results indicate that tinnitus patients have impaired cognitive performance overall, as measured by these variations of the Stroop paradigm, but hearing impairment cannot be excluded as a possible confounder. PMID- 11063239 TI - Effect of a single-channel wide dynamic range compression circuit on perception of stop consonant place of articulation. AB - Previous studies have shown that altering the amplitude of a consonant in a specific frequency region relative to an adjacent vowel's amplitude in the same frequency region will affect listeners' perception of the consonant place of articulation. Hearing aids with single-channel, fast-acting wide dynamic range compression (WDRC) alter the overall consonant-vowel (CV) intensity ratio by increasing consonant energy. Perhaps one reason WDRC has had limited success in improving speech recognition performance is that the natural amplitude balances between consonant and vowel are altered in crucial frequency regions, thus disturbing the aforementioned amplitude cue for determining place of articulation. The current study investigated the effect of a WDRC circuit on listeners' perception of place of articulation when the relative amplitude of consonant and vowel was manipulated. The stimuli were a continuum of synthetic CV syllables stripped of all place cues except relative consonant amplitudes. Acoustic analysis of the CVs before and after hearing aid processing showed a predictable increase in high-frequency energy, particularly for the burst of the consonant. Alveolar bursts had more high-frequency energy than labial bursts. Twenty-five listeners with normal hearing and 5 listeners with sensorineural hearing loss labeled the consonant sound of the CV syllables in unaided form and after the syllables were recorded through a hearing aid with single-channel WDRC. There were significantly more listeners who were unable to produce a category boundary when labeling the aided stimuli. Of those listeners who did yield a category boundary for both aided and unaided stimuli, there were significantly more alveolar responses for the aided condition. These results can be explained by the acoustic analyses of the aided stimuli. PMID- 11063240 TI - Speech, vocabulary, and the education of children using cochlear implants: oral or total communication? AB - This study examines the relationship between the teaching method, oral or total communication, used at children's schools and children's consonant-production accuracy and vocabulary development over time. Children who participated in the study (N = 147) demonstrated profound sensorineural hearing loss and had used cochlear implants for between 6 months and 10 years. Educational programs that used an oral communication (OC) approach focused on the development of spoken language, whereas educational programs that used a total communication (TC) approach focused on the development of language using both signed and spoken language. Using Hierarchical Linear Modeling (HLM) we compared the consonant production accuracy, receptive spoken vocabulary, and expressive spoken and/or signed vocabulary skills, over time, of children who were enrolled in schools that used either OC or TC approaches, while controlling for a number of variables. These variables included age at implantation, preoperative aided speech detection thresholds, type of cochlear implant device used, and whether a complete or incomplete active electrode array was implanted. The results of this study indicated that as they used their implants the children demonstrated improved consonant-production accuracy and expressive and receptive vocabulary over time, regardless of whether their school employed a TC or OC teaching method. Furthermore, there appeared to be a complex relationship among children's performance with the cochlear implant, age at implantation, and communication/teaching strategy employed by the school. Controlling for all variables, children in OC programs demonstrated, on average, superior consonant production accuracy, with significantly greater rates of improvement in consonant production accuracy scores over time compared to children in TC programs. However, there was no significant difference between OC and TC groups in performance or rate of growth in consonant-production accuracy when children received their implants before the age of 5 years. There was no significant difference between the OC and TC groups in receptive spoken vocabulary scores or in rate of improvement over time. However, children in the TC group achieved significantly higher receptive spoken vocabulary scores than children in the OC group if they received their implant before the age of 5 years. The TC group demonstrated superior scores and rates of growth on the expressive vocabulary measure (spoken and/or signed) when compared to the OC group if they received their implants during their preschool or early elementary school years. There was no significant difference if the children received their implants during middle elementary school. Regardless of whether children were in the OC or TC group, children who received their implants during preschool demonstrated stronger performance, on average, on all measures over time than children who received their implants during their elementary school years. The results of this study suggest that children may benefit from using cochlear implants regardless of the communication strategy/teaching approach employed by their school program and that other considerations, such as the age at which children receive implants, are more important. Implications and future research needs are discussed. PMID- 11063241 TI - Subjective vs. objective intelligibility of sentences in listeners with hearing loss. AB - The relation between rated intelligibility and correct key word repetitions of sentences was examined in listeners with normal hearing and listeners with hearing loss. Ten lists of 10 CID sentences were arranged in two randomly ordered groups. Listeners were asked to complete two tasks: (1) rate the intelligibility of 50 sentences on a scale of 0 to 100%, and (2) repeat each of the 50 sentences, which were scored as the number of key words repeated correctly. Sentences were presented at five signal-to-noise ratios. The start level for sentence presentation was established using the method of adjustment and the Revised Speech Intelligibility Rating passages. Correlations of rated intelligibility and correct repetition were 0.85 for listeners with normal hearing and 0.86 for listeners with hearing loss. This study confirms the earlier work of Speaks, Parker, Harris, and Kuhl (1972), and the findings demonstrate that listeners with hearing loss preserve the ability to rate the intelligibility of speech. PMID- 11063242 TI - Laryngeal factors in voiceless consonant production in men, women, and 5-year olds. AB - Voicing control in stop consonants has often been measured by means of voice onset time (VOT) and discussed in terms of interarticulator timing. However, control of voicing also involves details of laryngeal setting and management of sub- and supraglottal pressure levels, and many of these factors are known to undergo developmental change. Mechanical and aerodynamic conditions at the glottis may therefore vary considerably in normal populations as functions of age and/or sex. The current study collected oral airflow, intraoral pressure, and acoustic signals from normal English-speaking adults and children producing stop consonants and /h/ embedded in a short carrier utterance. Measures were made of stop VOTs, /h/ voicing and flow characteristics, and subglottal pressure during /p/ closures. Clear age and gender effects were observed for /h/: Fully voiced /h/ was most common in men, and /h/ voicing and flow data showed the highest variability among the 5-year-olds. For individual participants, distributional measures of VOT in /p t/ were correlated with distributional measures of voicing in /h/. The data indicate that one cannot assume comparable laryngeal conditions across speaker groups. This, in turn, implies that VOT acquisition in children cannot be interpreted purely in terms of developing interarticulator timing control, but must also reflect growing mastery over voicing itself. Further, differences in laryngeal structure and aerodynamic quantities may require men and women to adopt somewhat different strategies for achieving distinctive consonantal voicing contrasts. PMID- 11063243 TI - Lip muscle activity related to speech rate and loudness. AB - Changes in suprasegmental speech parameters may require adjustments in oral motor control that are reflected in the activity of perioral musculature. In order to evaluate possible patterns of difference, perioral surface electromyographic (EMG) signals were obtained from 20 adults who read a paragraph aloud at habitual rate and at self-judged proportionately slower and faster rates, at habitual loudness and at proportionately softer and louder levels, and in a "precise" manner. EMG amplitude analysis showed significant task effects, with higher average amplitudes for fast, loud, and precise speech and lower average amplitudes for slow and soft speech. These results are compatible with a model of multidimensional reorganization of speech motor control for suprasegmental changes applied to connected speech. PMID- 11063244 TI - Influence of continuous speaking on ventilation. AB - This study was conducted to explore the influence of speaking on ventilation. Twenty healthy young men were studied during periods of quiet breathing and prolonged speaking using noninvasive methods to measure chest wall surface motions and expired gas composition. Results indicated that all subjects ventilated more during speaking than during quiet breathing, usually by augmenting both tidal volume and breathing frequency. Ventilation did not change across repeated speaking trials. Quiet breathing was altered from its usual behavior following speaking, often for several minutes. Speaking-related increases in ventilation were found to be strongly correlated with lung volume expenditures per syllable. These findings have clinical implications for the respiratory care practitioner and the speech-language pathologist. PMID- 11063245 TI - Parental perceptions of children's communicative development at stuttering onset. AB - There has been clinical speculation that parents of young stuttering children have expectations of their children's communication abilities that are not well matched to the children's actual skills. We appraised the language abilities of 15 children close to the onset of stuttering symptoms and 15 age-, sex-, and SES matched fluent children using an array of standardized tests and spontaneous language sample measures. Parents concurrently completed two parent-report measures of the children's communicative development. Results indicated generally depressed performance on all child speech and language measures by the children who stutter. Parent report was closely attuned to child performance for the stuttering children; parents of nonstuttering children were less accurate in their predictions of children's communicative performance. Implications for clinical advisement to parents of stuttering children are discussed. PMID- 11063246 TI - Temporal and biomechanical characteristics of oropharyngeal swallow in younger and older men. AB - As the U.S. population ages, there is increasing need for data on the effects of aging in healthy elderly individuals over age 80. This investigation compared the swallowing ability of 8 healthy younger men between the ages of 21 and 29 and 8 healthy older men between the ages of 80 and 94 during two swallows each of 1 ml and 10 ml liquid. Videofluoroscopic studies of these swallows were analyzed to confirm the absence of swallowing disorders. Biomechanical analysis of each swallow was completed, from which data on temporal, range of motion, and coordination characteristics of the oropharyngeal swallow were taken. Position of the larynx at rest, length of neck, and pattern of hyoid bone movement were also compared between the two groups. None of the younger or older men exhibited any swallowing disorders. The C2 to C4 distance of older men was significantly shorter than that of younger men, and laryngeal position at rest was lower than in younger men but not significantly so. Older men had a significantly longer pharyngeal delay than younger men and significantly faster onset of posterior pharyngeal wall movement in relation to first cricopharyngeal opening. The older men exhibited significantly reduced maximum vertical and anterior hyoid movement as compared to the younger men even when accounting for the difference in C2 to C4 distance in older men. These data support the hypothesis of reduced muscular reserve in the swallows of older men as compared to younger men. Older men also exhibited less width of cricopharyngeal opening than younger men at 10 ml volume, indicating less upper esophageal sphincter flexibility in the swallows of older men. The potential for exercise to improve reserve is discussed. Significant changes in extent of hyoid elevation and duration of cricopharyngeal opening were seen as liquid bolus volume increased. PMID- 11063247 TI - Ataxic dysarthria. AB - Although ataxic dysarthria has been studied with various methods in several languages, questions remain concerning which features of the disorder are most consistent, which speaking tasks are most sensitive to the disorder, and whether the different speech production subsystems are uniformly affected. Perceptual and acoustic data were obtained from 14 individuals (seven men, seven women) with ataxic dysarthria for several speaking tasks, including sustained vowel phonation, syllable repetition, sentence recitation, and conversation. Multidimensional acoustic analyses of sustained vowel phonation showed that the largest and most frequent abnormality for both men and women was a long-term variability of fundamental frequency. Other measures with a high frequency of abnormality were shimmer and peak amplitude variation (for both sexes) and jitter (for women). Syllable alternating motion rate (AMR) was typically slow and irregular in its temporal pattern. In addition, the energy maxima and minima often were highly variable across repeated syllables, and this variability is thought to reflect poorly coordinated respiratory function and inadequate articulatory/voicing control. Syllable rates tended to be slower for sentence recitation and conversation than for AMR, but the three rates were highly similar. Formant-frequency ranges during sentence production were essentially normal, showing that articulatory hypometria is not a pervasive problem. Conversational samples varied considerably across subjects in intelligibility and number of words/ morphemes in a breath group. Qualitative analyses of unintelligible episodes in conversation showed that these samples generally had a fairly well-defined syllable pattern but subjects differed in the degree to which the acoustic contrasts typical of consonant and vowel sequences were maintained. For some individuals, an intelligibility deficit occurred in the face of highly distinctive (and contrastive) acoustic patterns. PMID- 11063248 TI - More on correct definition of the experimental unit: an extension of Max and Onghena (1999). PMID- 11063249 TI - The 2000 Chromosome Committee reports for the Mouse Genome. PMID- 11063250 TI - Twister mutant mice are defective for otogelin, a component specific to inner ear acellular membranes. AB - Deafness is a common sensory defect in human. Our understanding of the molecular bases of this pathology comes from the study of a few genes that have been identified in human and/or in mice. Indeed, deaf mouse mutants are good models for studying and identifying genes involved in human hereditary hearing loss. Among these mouse mutants, twister was initially reported to have abnormal behavior and thereafter to be deaf. The recessive twister (twt) mutation has been mapped on mouse Chromosome (Chr) 7, homologous to the long arm of human Chr 15 (15q11). Otog, the gene encoding otogelin, a glycoprotein specific to all the acellular membranes of the inner ear, is also localized to mouse Chr 7, but in a region more proximal to the twister mutation, homologous to the short arm of human Chr 11 (11p15) carrying the two deafness loci, DFNB 18 and USH C. Mutant mice resulting from the knock-out of Otog, the Otogtm1Prs mice, present deafness and severe imbalance. Although twt had been mapped distally to Otog, these data prompted us to test whether twt could be due to a mutation in the Otog locus. Here, we demonstrate by genetic analysis that twt is actually allelic to Otogtm1Prs. We further extend the phenotypical analysis of twister mice, documenting the association of a severe vestibular phenotype and moderate to severe form of deafness. Molecular analysis of the Otog gene revealed the absence of detectable expression of Otog in the twister mutant. The molecular and phenotypical description of the twt mouse mutation, Otogtwt, reported herein, highlights the importance of the acellular membranes in the inner ear mechanotransduction process. PMID- 11063251 TI - Localization of the mouse kidney disease (kd) gene to a YAC/BAC contig on Chromosome 10. AB - Mice that are homozygous for the kidney disease (kd) gene on Chromosome (Chr) 10 spontaneously develop a progressive and fatal interstitial nephritis. The disease phenotype is similar to that of the human disease, juvenile nephronophthisis. Using a backcross and intercross breeding strategy and analysis of over 900 resultant progeny, this genetic locus has now been mapped to a minimal co segregating region of approximately two megabases between D10Mit 193 and D10Mit 38. The location assigned to kd by this study is over 3 cM from the current Mouse Genome Database location. The entire interval has been cloned in yeast artificial chromosome (YAC) and bacterial artificial chromosome (BAC) clones. Recombinant analysis has permitted assignment of 13 Mit microsatellite markers to positions near or within the region. Two new markers have been identified by using single strand conformation polymorphism (SSCP) analysis of sequenced BAC ends. Several BAC end sequences align with human BAC clones from Chr 6q2 that contain NR2E1. Snx3, and Ros1. Three murine genes, CD24a, fyn, and ColX reported to map in or near the kd region as defined by this study have been evaluated. Though not definitely excluded, they appear to be unlikely candidates. PMID- 11063252 TI - The hormone-sensitive lipase gene is transcribed from at least five alternative first exons in mouse adipose tissue. AB - Hormone-sensitive lipase (HSL) mediates triglyceride hydrolysis in adipocytes, in which its expression varies with physiological stress and is controlled posttranslationally and transcriptionally. We sequenced the mouse HSL gene for 8.2 kb upstream of the translation start codon and studied the steady-state HSL mRNA levels in mouse adipose tissue. In 50 clones derived from primer extension and PCR of mouse adipose cDNA, we found five distinct 5' extremities that correspond to distinct exons in genomic DNA. Exon A is located approximately 7 kb 5' to the HSL translation start site. Exons B. C, and D are clustered 1.5-2 kb upstream, and the previously described exon 1 is immediately upstream and contiguous with the previously described HSL translation start site. Exon A is located -7 kb upstream and contains an in-frame methionine codon that could potentially generate another HSL isoform with 43 additional N-terminal residues. cDNA clones containing the newly described exons suggested that each exon has several transcription start sites but that all splice to an acceptor site located 20 nt upstream of the translation initiation codon in exon 1. HSL transcription in mouse adipose tissue originates from multiple sites in the 7-kb region between exon A and exon 1, with peaks at exon C (50-70% of HSL transcripts), exon 1 (5 30%), and exon A (approximately 10%). There are multiple potential transcription factor-binding elements upstream of each exon, suggesting the possibility of differential transcriptional regulation of HSL in different tissues and under various physiologic conditions. PMID- 11063253 TI - Linkage disequilibrium and haplotype mapping of a skin cancer susceptibility locus in outbred mice. AB - Car-R (carcinogenesis-resistant) and Car-S (carcinogenesis-susceptible) outbred mice, obtained by phenotypic selection from an initial intercross of eight inbred strains, show a >100-fold difference in their susceptibility to two-stage skin tumorigenesis. We found that the lines carry a high degree of genetic polymorphism. with an average heterozygosity of 0.39. This polymorphism allowed the use of linkage disequilibrium (LD) and haplotype analysis for the mapping of a skin cancer modifier locus on Chr 7, in a short region of 6 cM, around the Tyr gene. Car-S mice inherited the susceptibility allele at this locus from the A/J, BALB/c, SJL/J, and SWR/J strains. Our results demonstrate the feasibility and usefulness of mapping disease genes by LD in phenotypically selected, genetically heterogeneous animals. PMID- 11063254 TI - Construction of a high-throughput rat genetic mapping system with 466 arbitrarily primed-representational difference analysis markers. AB - Linkage mapping of quantitative trait loci (QTLs) requires genetic markers that can be efficiently genotyped for a large number of individuals. To isolate genetic markers suitable for this purpose, we previously established the arbitrarily primed RDA (AP-RDA) method. Dot-blotting AP-PCR products (AP amplicons) onto filters at a high density and hybridization of the filters with the AP-RDA markers made it possible to genotype a large number of individuals simultaneously for multiple loci. In this study, by using 25 primers or primer combinations, we isolated a total of 419 AP-RDA markers by subtracting the AP amplicon of BUF rats from that of ACI rats, and vice versa. By combining 47 previously isolated markers, a rat genetic map was drawn with 466 AP-RDA markers. Between two given strains of rats other than ACI and BUF, the average informativeness of the markers was 38%. As for the intercross of ACI and BUF rats, 12 selected primers served to genotype 259 loci. In addition, the amounts and quality of genomic DNA to be used for AP-PCR were examined to guarantee reliable genotyping. Now, initial genome scanning of the rat for linkage analysis can be performed efficiently using this mapping system with AP-RDA markers. PMID- 11063255 TI - Analysis and origins of the human and mouse RNase L genes: mediators of interferon action. AB - The 2',5'-oligoadenylate-activated enzyme, RNase L, is an endoribonuclease implicated in the antiviral and apoptotic activities of interferons. To probe the genetics of the 2-5A system, the human and mouse genes were cloned, characterized, and compared. The first coding exon of both genes encodes the regulatory regions of RNase L, 67-70% of the proteins including nine ankyrin repeats, the 2-5A binding domain, and several protein kinase homology motifs. In contrast, the coding sequence for the ribonuclease domain in the mouse and human gene is divided among three exons. The transcriptional start site of the human RNase L gene was located in noncoding exon I by primer extension analysis. A complete coding sequence of mouse RNase L was obtained revealing a 735-amino acid protein with 64% identity to human RNase L. A hypothesis is presented concerning the evolutionary relationship of RNase L to both an ankyrin repeat protein kinase and the kinase-endoribonuclease. IRE1, that mediates the unfolded protein response. PMID- 11063256 TI - Comparative mapping of human Chromosome 14q11.2-q13 genes with mouse homologous gene regions. AB - An examination of the synteny blocks between mouse and human chromosomes aids in understanding the evolution of chromosome divergence between these two species. We comparatively mapped the human (HSA) Chromosome (Chr) 14q11.2-q13 cytogenetic region with the intervals of orthologous genes on mouse (MMU) chromosomes. A lack of conserved gene order was identified between the human cytogenetic region and the interval of orthologs on MMU 12. The evolutionary breakpoint junction was defined within 2.5 Mb, where the conserved synteny of genes on HSA 14 changes from MMU 12 to MMU 14. At the evolutionary breakpoint junction, a human EST (GI: 1114654) with identity to the human and mouse BCL2 interacting gene, BNIP3, was mapped to mouse Chr 3. New gene homologs of LAMB1, MEOX2, NRCAM, and NZTF1 were identified on HSA 7 and on the proximal cytogenetic region of HSA 14 by mapping mouse genes recently reported to be genetically linked within the relevant MMU 12 interval. This study contributes to the identification of homology relationships between the genes of HSA 14q11.2-q13 and mouse Chr 3, 12, and 14. PMID- 11063257 TI - Human PRRX1 and PRRX2 genes: cloning, expression, genomic localization, and exclusion as disease genes for Nager syndrome. AB - In this study, we extend our examination of the function of the Prrx1 (a.k.a Mhox, Prx1, K-2, and Pmx1) as well as Prrx2 (a.k.a. S8 and Prx2) genes by characterizing the expression of the human orthologs and their potential for causing specific human malformations. The expression pattern of PRRX2 and its close relative, PRRX1, were analyzed in human tissue by RT-PCR. Although the expression of these human genes is similar to their mouse orthologs, there are notable differences in expression. PRRX2 was detected in the human kidney and lung, whereas in mice and chickens neither of these tissues has been reported to express Prrx2. For PRRX1 the expression pattern was quite similar to other vertebrates, but the ratio of the two isoforms was reversed. To begin the search for the gene-disease connection, both genes were mapped to human chromosomes by FISH. The PRRX1 locus maps to 1q23, whereas the PRRX2 locus maps to 9q34.1. This localization, along with the recently described phenotypes of the gene-targeted Prrx1, Prrx2 and double mutant mice, enabled us to search the human disease databases for similar malformations. This examination suggested that mutations at the PRRX1 and/or PRRX2 loci could result in Nager Acrofacial Dysostosis (NAFD) syndrome. We obtained DNA samples from eight patients with NAFD, as well as two patients with Miller syndrome, and analyzed them for mutations in the PRRX1 and PRRX2 genes. The data excludes mutations in the presumed coding sequences of these genes from causing NAFD. PMID- 11063258 TI - HIP12 is a non-proapoptotic member of a gene family including HIP1, an interacting protein with huntingtin. AB - Huntingtin-interacting protein I (HIP1) is a membrane-associated protein that interacts with huntingtin, the protein altered in Huntington disease. HIP1 shows homology to Sla2p, a protein essential for the assembly and function of the cytoskeleton and endocytosis in Saccharomyces cerevisiae. We have determined that the HIP1 gene comprises 32 exons spanning approximately 215 kb of genomic DNA and gives rise to two alternate splice forms termed HIP1-1 and HIP1-2. Additionally, we have identified a novel protein termed HIP12 with significant sequence and biochemical similarities to HIP1 and high sequence similarity to Sla2p. HIP12 differs from HIP1 in its pattern of expression both at the mRNA and protein level. However, HIP1 and HIP12 are both found within the brain and show a similar subcellular distribution pattern. In contrast to HIP1, which is toxic in cell culture, HIP12 does not confer toxicity in the same assay systems. Interestingly, HIP12 does not interact with huntingtin but can interact with HIP1. suggesting a potential interaction in vivo that may influence the function of each respective protein. PMID- 11063259 TI - The olfactory receptor gene superfamily: data mining, classification, and nomenclature. AB - The vertebrate olfactory receptor (OR) subgenome harbors the largest known gene family, which has been expanded by the need to provide recognition capacity for millions of potential odorants. We implemented an automated procedure to identify all OR coding regions from published sequences. This led us to the identification of 831 OR coding regions (including pseudogenes) from 24 vertebrate species. The resulting dataset was subjected to neighbor-joining phylogenetic analysis and classified into 32 distinct families, 14 of which include only genes from tetrapodan species (Class II ORs). We also report here the first identification of OR sequences from a marsupial (koala) and a monotreme (platypus). Analysis of these OR sequences suggests that the ancestral mammal had a small OR repertoire, which expanded independently in all three mammalian subclasses. Classification of "fish-like" (Class I) ORs indicates that some of these ancient ORs were maintained and even expanded in mammals. A nomenclature system for the OR gene superfamily is proposed, based on a divergence evolutionary model. The nomenclature consists of the root symbol 'OR', followed by a family numeral, subfamily letter(s), and a numeral representing the individual gene within the subfamily. For example, OR3A1 is an OR gene of family 3, subfamily A, and OR7E12P is an OR pseudogene of family 7, subfamily E. The symbol is to be preceded by a species indicator. We have assigned the proposed nomenclature symbols for all 330 human OR genes in the database. A WWW tool for automated name assignment is provided. PMID- 11063260 TI - Genome scan identifies a locus affecting gamma-globin level in human beta-cluster YAC transgenic mice. AB - Genetic factors affecting postnatal gamma-globin expression--a major modifier of the severity of both beta-thalassemia and sickle cell anemia--have been difficult to study. This is especially so in mice, an organism lacking a globin gene with an expression pattern equivalent to that of human gamma-globin. To model the human beta-cluster in mice, with the goal of screening for loci affecting human gamma-globin expression in vivo, we introduced a human beta-globin cluster YAC transgene into the genome of FVB/N mice. The beta-cluster contained a Greek hereditary persistence of fetal hemoglobin (HPFH) gamma allele, resulting in postnatal expression of human gamma-globin in transgenic mice. The level of human gamma-globin for various F1 hybrids derived from crosses between the FVB/N transgenics and other inbred mouse strains was assessed. The gamma-globin level of the (C3HeB/FeJ x FVB/N)F1 transgenic mice was noted to be significantly elevated. To map genes affecting postnatal y-globin expression, we performed a 20 centiMorgan (cM) genome scan of a (C3HeB/FeJ x FVB/N)F1 transgenics x FVB/N backcross, followed by high-resolution marker analysis of promising loci. From this analysis we mapped a locus within an 18-cM interval of mouse Chromosome (Chr) 1 (LOD = 4.3) that contributes 10.9% of variation in gamma-globin level. Combining transgenic modeling of the human beta-globin gene cluster with quantitative trait analysis, we have identified and mapped a murine locus that impacts on human gamma-globin level in vivo. PMID- 11063261 TI - cDNA cloning, chromosomal mapping, and expression analysis of human VE-Cadherin 2. AB - Murine vascular endothelial cadherin-2 (VE-cad-2) is a cellular adhesion molecule that is distinct from vascular endothelial cadherin 1 (VE-cad-1) in that it does not interact with catenins and does not appear to affect cell migration or growth. In this study, we have cloned a full-length cDNA of the human homolog of VE-cad-2 and used it to map the chromosomal locus of the VE-cad-2 gene. Human VE cad-2 maps to Chromosome (Chr) 5q31. The cDNA of human VE-cad-2 is highly homologous to mouse VE-cad-2, except for a C-terminal tail. The genomic structure of VE-cad-2 is strikingly similar to that reported for a large family of neuronal protocadherin genes mapped to Chr 5q, yet the amino acid sequences between VE-cad 2 and the protocadherins are substantially divergent. The promoter of human VE cad-2 contains two TATA boxes and transcription initiates from a single site 3' to these elements. Similar to mouse VE-cad-2, the human gene is expressed primarily in highly vascularized tissues. PMID- 11063262 TI - The gene encoding rat 3-phosphoglycerate dehydrogenase. PMID- 11063264 TI - Do no harm. PMID- 11063263 TI - Cloning, structure, expression analysis, and assignment to mouse chromosome 7 of the gene Zfp296 encoding a zinc finger protein. PMID- 11063265 TI - Benefits of drinking polyphenols. PMID- 11063266 TI - The most embarrassing symptoms. PMID- 11063267 TI - Concepts of traditional Oriental medicine. AB - BACKGROUND: Traditional Oriental medicine is taught and practiced in many parts of the world, including the United States. It is based on centuries-old philosophies of disease causality and on empirical validation of diagnostic and treatment procedures. This article presents a perspective on traditional Oriental medicine. METHODS: The traditional Oriental medicine physician considers the body to normally be in a state of balance or harmony that can be disrupted by certain pernicious influences. These influences affect various body systems in ways that can be detected using a four-part examination, which includes looking, listening and smelling, asking, and touching. During the examination, special emphasis is placed on analyzing the rate and rhythm of the pulse. RESULTS: Illness results when there is disharmony within the body. Therefore, the goal of the Oriental medicine physician is to restore harmony in the patient. This can be accomplished by using techniques such as acupuncture, moxibustion, and administration of herbal preparations. CONCLUSIONS: The underlying assumptions, diagnostic techniques, and treatments used by traditional Oriental medicine physicians can seem strange to Western-trained physicians, but they have proved efficacious for the treatment of patients over many millennia. PMID- 11063268 TI - U.S. clinical investigation of the Artisan myopia lens for the correction of high myopia in phakic eyes. Report of the results of phases 1 and 2, and interim phase 3. AB - PURPOSE: The purpose of this article is to present the results of the U.S. Clinical investigation of the Artisan anterior chamber iris fixed lens implant for the correction of myopia in phakic eyes. METHODS: A prospective, multi center, FDA-supervised trial was designed and undertaken in the United States by Ophtec USA, Inc., Boca Raton, Florida to determine the safety and efficacy of the Artisan lens as a potential refractive treatment for patients with high myopia. During the trial, two different models of the Artisan lens were used: one with a 5-mm optical zone (model 206) and the other with a 6-mm optical zone (model 204). RESULTS: The data presented comprise 176 enrolled subjects and 264 implant procedures. The most frequently chosen Artisan lens power was -13.00 D (average, 12.76 D; SD, 3.24). The postoperative results at 6 months for all eyes (n = 135) showed 100% of patients were 20/40 or better best-corrected while 72% gained one or more lines and 22% gained two or more lines regardless of degree of astigmatism or postoperative goal. Through the course of the study, intraocular pressure maintained a level with a mean below 16 mmHg. In general, total reported complications in the patient cohort decreased over time, dropping from 39% at the initial visit to 10% at visit four and 0% at visit seven. CONCLUSIONS: On the basis of the interim results of the U.S. Clinical Investigation of the Artisan Myopia Lens for the Correction of High Myopia in Phakic Eyes, the Artisan anterior chamber phakic IOL may offer an option for correction of high degrees of myopia. Refractive outcomes were exceptional and complications were minimal and amenable to treatment. PMID- 11063269 TI - Visual-field defects in well-defined retinal lesions using Humphrey and Dicon perimeters. AB - BACKGROUND: This study was undertaken to correlate field defects in patients having well-defined retinal lesions within the central 30 degrees using Humphrey and Dicon perimetry standard threshold programs and to compare these results against the standard of fundus photography. METHODS: Eleven eyes of 11 patients had various well-defined retinal lesions. Ten of those subjects had best corrected visual acuity of 20/25 or better. Subjects had either one previous field or no previous experience with any automated perimeter and were tested using either the 30-2 Full Threshold or SITA Standard program of the Humphrey Field Analyzer (HFA) Model 750 and the Central Grid Threshold program (#9) (a full-threshold program) of the Dicon LD-400 perimeter. Defects were scored and compared with fundus photographs scored by an independent observer. RESULTS: Significant high positive correlation coefficients were found between Humphrey, Dicon, and fundus photography, for mean number of significant field defects and for mean deviations. There were no significant differences between the mean scores, mean deviations, or fixation loss percentages. The time the patient occupied a perimeter was 56% less with the Dicon perimeter than with the Humphrey perimeter when the Full Threshold test was used and 8% less when the SITA Standard was used. CONCLUSION: In this study of patients with well-defined retinal lesions, both Humphrey and Dicon Central Threshold field programs exhibited an equally high positive correlation in mapping scotomas of expected size and depth when compared with fundus photography, with no significant difference in fixation loss frequency. However, the total testing time was less with Dicon perimetry than with Humphrey perimetry. PMID- 11063270 TI - Air quality and ocular discomfort aboard commercial aircraft. AB - BACKGROUND: Aircraft cabin air quality has been a subject of recent public health interest. Aircraft environments are designed according to standards to ensure the comfort and well-being of the occupants. The upper and lower limits of humidity set by ASHRAE standards are based on the maintenance of acceptable thermal conditions established solely on comfort considerations, including thermal sensation, skin wetness, skin dryness, dry eyes and ocular discomfort. The purpose of this study is to investigate the influence of air (carbon dioxide level, relative humidity, and temperature) aboard commercial aircraft on ocular discomfort and dry eye of aircraft personnel and passengers. METHODS: Measurements of indoor air quality were performed in 15 different aircraft at different times and altitudes. Forty-two measurements of carbon dioxide, temperature, and humidity were performed with portable air samplers every 5 minutes. Passenger loads did not exceed 137 passengers. RESULTS: Thermal comfort rarely met ASHRAE standards. Low humidity levels and high carbon dioxide levels were found on the Airbus 320. The DC-9 had the highest humidity level and the Boeing-767 had the lowest carbon dioxide level. CONCLUSIONS: Air quality was poorest on the Airbus 320 aircraft. This poor level of air quality may cause intolerance to contact lenses, dry eyes, and may be a health hazard to both passengers and crew members. Improved ventilation and aircraft cabin micro environments need to be made for the health and comfort of the occupants. PMID- 11063272 TI - The Internet as a practice tool. PMID- 11063271 TI - Nutritional influences on eye health. AB - BACKGROUND: As with cardiovascular disease, nutrition and environmental intervention present new clinical practice and research opportunities in eye care. This includes maximizing health potential with food choice, the use of supplementation in high-risk and noncompliant patients, avoiding toxins such as cigarettes, and encouraging exercise. METHOD: The author provides his perspective on viewing biochemical, physiological, and epidemiological research in the context of disease prevention in both common and uncommon applications. In most cases, this approach requires food frequency intake evaluation and application of physiological optics to measure efficacy when there is an alteration in diet and/or addition of supplementation. These are details of ocular integrative medical practice that are ignored or underutilized. RESULTS: There are multiple opportunities for intervention in the age-related diseases of cataract, macular degeneration, and glaucoma. A summary of the application of nutrition to create a model for treatment of age-related macular degeneration is presented. CONCLUSIONS: Economic incentives, now lacking, are required to explore new paradigms in nutritional preventative eye care. Schools and colleges of Optometry could play a vital public health role. PMID- 11063273 TI - Don't overlook advertising co-op. PMID- 11063274 TI - Management 101: or what management theory can teach optometrists. PMID- 11063275 TI - Childhood asthma hospitalizations--King County, Washington, 1987-1998. AB - Since 1980, asthma prevalence, hospitalization, and mortality have been increasing in the United States (1). Because of concern about asthma morbidity in children in King County, Washington (2), Public Health-Seattle and King County (PH-SKC) conducted a study that analyzed trends in local hospitalizations for childhood asthma during 1987-1998. This report summarizes the results of this analysis, which indicate that the youngest children and the poorest communities have the highest rates of asthma hospitalization. PMID- 11063276 TI - Self-reported concern about food security--eight states, 1996-1998. AB - Food security is defined as having access at all times to enough food for an active, healthy lifestyle (1,2). This definition implies that safe and nutritious foods are available and that household resources are sufficient to meet cost. Recognition that hunger and food security are problems in the United States led to the development and implementation of measures of hunger and food security on national surveys. One of the national health objectives for 2010 is to increase food security and reduce the risk for hunger among all households (objective 19 18) (1). To characterize state-level prevalence of concern about food security, data were analyzed for the eight states that used the Social Context Module of the Behavioral Risk Factor Surveillance System (BRFSS) during 1996-1998. This report summarizes the results of this analysis and indicates that approximately 4%-6% of adults reported a concern about having enough food for themselves or their family during the preceding month. PMID- 11063277 TI - Hospital-based policies for prevention perinatal Group B streptococcal disease- United States, 1999. AB - Group B streptococcus (GBS) is the leading cause of sepsis, meningitis, and pneumonia in newborns in the United States (1). Because intrapartum prophylactic antibiotics reduce mother-to-child GBS transmission (2), in 1996, CDC, the American College of Obstetricians and Gynecologists, and the American Academy of Pediatrics recommended that hospitals adopt formal GBS prevention policies (2-4). From 1994 to 1997, the proportion of hospitals with formal intrapartum GBS prevention policies increased from 39% to 59% (5,6); hospitals that implemented policies reported less GBS disease among neonates (7). In 1999, CDC's Active Bacterial Core Surveillance (ABCs) system surveyed hospitals in eight states about their GBS prevention policies. This report summarizes the results of that analysis and indicates that in 1999, the proportion of hospitals with formal policies had not changed since 1997; however, a higher proportion of hospitals have implemented measures to improve policy compliance. PMID- 11063278 TI - The theory of treating Type 2 diabetes. AB - Type 2 diabetes mellitus is a chronic, progressive disease affecting many millions of people worldwide. It carries a great burden of morbidity and premature mortality for the individual, and places great demands on healthcare systems and resources. We now know from clinical studies that improved control of Type 2 diabetes can to some degree reduce its burden. We also know that in the context of a clinical trial, the treatments available to us can do much to improve control in many patients (although all will fall short of 'normality'). International guidelines for management of Type 2 diabetes, quite correctly, encourage us to strive for levels of control where we believe the risk of complications is lowest. But is this happening in everyday practice? Data from a survey in three countries show that there is a great difference between the theory of diabetes care and the reality of clinical practice, with levels of glycaemic control in most patients falling short of desired levels. A consideration of the pathophysiology of Type 2 diabetes reveals that it is a complex syndrome focussing on the progressive failure of the pancreatic beta cell. By acknowledging this fact, and addressing our therapeutic efforts appropriately, we may help to span the gap between theory and reality. PMID- 11063279 TI - The ADDITION study: proposed trial of the cost-effectiveness of an intensive multifactorial intervention on morbidity and mortality among people with Type 2 diabetes detected by screening. AB - OBJECTIVE: The overall aims of the ADDITION study are to evaluate whether screening for prevalent undiagnosed Type 2 diabetes is feasible, and whether subsequent optimised intensive treatment of diabetes, and associated risk factors, is feasible and beneficial. DESIGN: Population-based screening in three European countries followed by an open, randomised controlled trial. SUBJECTS AND METHODS: People aged 40-69 y in the community, without known diabetes, will be offered a random capillary blood glucose screening test by their primary care physicians, followed, if equal to or greater than 5.5 mmol/l, by fasting and 2-h post-glucose-challenge blood glucose measurements. Three thousand newly diagnosed patients will subsequently receive conventional treatment (according to current national guidelines) or intensive multifactorial treatment (lifestyle advice, prescription of aspirin and ACE-inhibitors, in addition to protocol-driven tight control of blood glucose, blood pressure and cholesterol). Patients allocated to intensive treatment will be further randomised to centre-specific interventions to motivate adherence to lifestyle changes and medication. Duration of follow-up is planned for 5 y. Endpoints will include mortality, macrovascular and microvascular complications, patient health status and satisfaction, process-of care indicators and costs. PMID- 11063280 TI - Barriers to good glycaemic control: the patient's perspective. AB - Diabetes currently affects at least 120 million people worldwide, and this figure is rising steadily. Intensive treatment improves outcome in terms of morbidity from late diabetic complications and quality of life, but in order for patients to reap such benefits, they must commit to major, long-term changes in lifestyle. The physician's concept of diabetes is often very different from the patient's; and the implementation of a treatment plan acceptable to both is only possible when open communication fosters discussion and patient autonomy, and treatment is seen as logical, acceptable and feasible within the daily life of each patient. Barriers that impair patients' ability to achieve good glycaemic control include those relating to lifestyle, education, psychology and their environment. An appreciation of barriers to good glycaemic control from the patient's perspective underlies the ability to minimise obstacles and improve outcome in terms of quality of life and metabolic control. PMID- 11063281 TI - Review of prandial glucose regulation with repaglinide: a solution to the problem of hypoglycaemia in the treatment of Type 2 diabetes? AB - Type 2 diabetes mellitus is characterised by abnormal beta-cell function (present at the time of diagnosis) that is often associated with insulin resistance. An important and consistent pathophysiological finding is the failure to produce adequate increments in insulin secretion in response to carbohydrate intake. Therefore, insulin secretagogue therapy, particularly when focused on prandial glucose regulation, is a logical approach to treatment because it addresses one of the most fundamental pathophysiological aspects of the disease. However, the traditional secretagogues-the sulphonylureas--have long been associated with the unwanted effect of hypoglycaemia. This is particularly likely to occur when drugs with lengthy plasma half-lives, prolonged drug-receptor interactions, active metabolites or a reliance on renal clearance are used. The problem is most prevalent in elderly patients, where sulphonylurea-induced hypoglycaemia may be related to failure to comply with strict mealtimes or the need for supplementary food intake, often in the context of compromised renal function. Data from large scale outcome studies demonstrate that when tight glycaemic control is achieved through aggressive antidiabetic therapy, late diabetic complications can be significantly reduced. However, the pursuit of stricter HbA1c targets with more aggressive interventions may increase the risk of hypoglycaemia. This is an irony because the clinical need to avoid hypoglycaemia and patients' apprehension of it present barriers to the achievement of beneficial glycaemic targets. However, an increased risk of hypoglycaemia may not be inevitable with insulin secretagogue therapy. The recently introduced carbamoylmethyl benzoic acid derivative, repaglinide, has pharmacological properties that are well suited to its intended role as a prandial glucose regulator. When taken prior to main meals, the rapid onset and relatively short duration of action of repaglinide aid disposal of the mealtime glucose load, without continued stimulation of pancreatic beta-cells in the postprandial fasting period. Repaglinide is also characterised by hepatic metabolism and elimination, which is an advantage in the context of impaired renal function. Prandial glucose regulation with repaglinide selectively increases insulin secretion, and hence limits glucose excursions, in the prandial phase. If a meal is omitted, so too is the corresponding dose. This more flexible approach to the management of Type 2 diabetes has a number of advantages when compared with the fixed daily dosing regimens of sulphonylureas, among them a reduced risk of hypoglycaemia--a benefit that is particularly marked in the context of missed or irregular meals. PMID- 11063282 TI - The importance of early insulin secretion and its impact on glycaemic regulation. AB - Type 2 diabetes is characterised by a progressive deterioration of the prandial insulin response, in a situation of continuing insulin resistance. Early phase insulin release is attenuated and delayed and there is a consequent failure to suppress glucagon secretion and curtail hepatic glucose production and gluconeogenesis. Postprandial plasma glucose concentration rises to pathological levels and fails to return to normal before the patient consumes their next meal, creating a problem of continuous daytime hyperglycaemia. Although late insulin secretion is preserved it does not rectify the hyperglycaemia. The pathology of excessive prandial glucose excursions and continual daytime hyperglycaemia can be normalised, at least in part, if early-phase insulin availability is restored through pharmacologic intervention. Initially, the feasibility of this approach was demonstrated experimentally with the use of carefully controlled insulin infusions or insulin analogue injections. More recently, the availability of the rapid or early augmentor of insulin secretion--repaglinide--provides a means for restoring prandial glucose regulation with oral therapy. Placebo-controlled and oral hypoglycaemic agent (OHA) comparative studies of repaglinide have established its antidiabetic efficacy and flexible mealtime/dosing studies have confirmed the importance of the prandial approach to treatment. Prandial glucose regulation with repaglinide has also been demonstrated to provide synergies when used as combination therapy with insulin sensitising agents. As a strategy, prandial glucose regulation has a number of theoretical advantages over the use of fixed doses of conventional insulin secretagogues, and these have been borne out in clinical trials. As well as offering a more flexible approach to treatment, prandial repaglinide is associated with a reduced risk of severe hypoglycaemia. PMID- 11063283 TI - Prandial glucose regulation with repaglinide: its clinical and lifestyle impact in a large cohort of patients with Type 2 diabetes. AB - OBJECTIVE: Prandial glucose regulation has the potential for achieving good metabolic control with a low risk of hypoglycaemia and increased flexibility with regard to eating patterns. Comparative studies have suggested that the prandial glucose regulator repaglinide is at least equivalent to sulphonylureas in terms of efficacy, but incurs a lower risk of major hypoglycaemia. However, these trials employed fixed dosing and mealtime regimens, so repaglinide was not used as intended. This prospective investigation in a daily clinical setting aimed to assess the efficacy and tolerability profile of flexible prandial glucose regulation with repaglinide in Type 2 diabetes. DESIGN: 5,985 patients with Type 2 diabetes in Germany were surveyed prospectively. These patients were assessed before and after a mean of 46 days treatment with repaglinide. At baseline, available data showed that 64% of patients had previously received therapy with conventional oral antidiabetic drugs, 22% were on diet alone, and 13% were naive to any treatment. RESULTS: Overall, mean HbA1c decreased from 8.6 to 7.4%, fasting blood glucose from 183.9 to 134.2 mg/dl (10.2 to 7.4 mmol/l), blood glucose prior to main meals from 198.5 to 141.4 mg/dl (11 to 7.8 mmol/l), and blood glucose 2 hours after main meals from 219.3 mg/dl to 153.2 mg/dl (12.2 to 8.5 mmol/l). Subgroup analysis showed significant improvements in each of these parameters (P<0.0001) in therapy-naive patients, in patients switched from other oral antidiabetic drugs, and in patients receiving repaglinide as combination therapy. Body weight decreased slightly (1.2+/-2.7 kg). Only 49 hypoglycaemic episodes were reported, of which 38 cases were mild and no adverse sequelae to these events have been reported. Repaglinide also led to a liberating effect on lifestyle when patients were switched from other oral hypoglycaemic agents (OHAs), with 80% reporting a sense of relief at the prospect of being able to miss meals. The proportion of these patients reporting lifestyle restrictions as a result of fixed mealtimes declined from 36% to 7%. Before switching, 38% of the patients admitted to eating when not hungry for fear of hypoglycaemia, but only 10% continued this behaviour and patients took fewer supplementary snacks after switching to repaglinide. CONCLUSION: Prandial glucose regulation with repaglinide improves metabolic control in patients with Type 2 diabetes without causing weight gain and with few hypoglycaemic episodes. This beneficial effect is seen in patients who are therapy-naive, have switched from alternative OHAs, or are in need of combination therapy. The prandial approach to treatment has a liberating effect with regard to eating behaviour that is welcomed by most patients switched from alternative therapies. PMID- 11063284 TI - The influence of autonomy and paternalism on communicative behaviors in mother daughter relationships prior to dependency. AB - Parents and their adult children rarely have discussions regarding caregiving preferences, especially before the onset of dependency. Families develop decision making practices during caregiving, ideally, ones that maintain the care recipient's autonomy. Maintaining autonomy is essential because limiting autonomy leads to potentially negative health consequences. This study examined the attitudes of older, independent mothers and their adult daughters (n = 36 dyads) as well as their communication behaviors during decision making. All the mothers and daughters (100%) held strong beliefs in shared autonomy. Mothers (55.6%) and daughters (58.3%) were about evenly split in their strength of belief in independent autonomy. More mothers (63.9%) held strong beliefs in paternalism than did daughters (36.1%). In their conversations, daughters talked more when the mother and daughter held stronger beliefs in paternalism. These findings suggest that an individual's attitudes toward paternalism influence who controls conversations between parents and their adult children regarding caregiving, even before the parent has begun to show any signs of dependency. Once caregiving begins, it can be too late to change already ingrained patterns of decision making. Although this study takes a step toward establishing an understanding of how families develop decision-making processes utilized during caregiving, the sample size and composition limit generalizability. Future studies should follow families, as the parents make the transition from independence to dependence, to develop a better understanding of the factors involved in successfully making such a critical transition in the family's life. PMID- 11063285 TI - Predictors of participation in health care at menopause. AB - Patient participation in health care is widely advocated but little is known about the factors influencing patients' active participation. To learn whether attitudes and beliefs helped predict patient participation, 252 midlife women completed measures of self-efficacy, perceived barriers to participation, attitudes, subjective norms, and intentions following a decision to support intervention on menopause. Two months following the intervention, 63 women who had visited a health care provider returned questionnaires by mail regarding self reported participation in the health care encounter and satisfaction with the decision about estrogen replacement therapy. The results were that self-efficacy (beta = .19; p < .05) significantly predicted intention to participate in the next health care encounter. Both patient intentions (beta = .40;p < .05) and perceived barriers (beta = -.27; p < .05) were significant predictors of self reported participation. Active participation (beta = .42;p < .05) and attitude (beta =.25;p < .05) were associated with increased satisfaction with decision. Participation resulted in increased self-efficacy (beta = .30;p < .05). The conclusions were that midlife women who express more confidence in their ability to participate in their health care have greater intentions to participate. Women who experience fewer barriers to participation and have more intention to participate are more likely to report actively participating in a health care encounter, and self-reported participation and more positive attitudes toward participation are associated with greater satisfaction with decisions. PMID- 11063286 TI - Family caregivers' use of humor in conveying information about caring for dependent older adults. AB - Twenty-three family caregivers were interviewed using a semistructured interview format to explore their experiences managing medication administration and providing long-term care to frail elderly family members. Content analysis of the transcripts utilized the arousal-relief theory of humor (Berlyne, 1969) to understand the frequent use of humorous anecdotes found throughout the interviews. For example, caregivers of individuals experiencing dementia often included smiles, jokes, and "punch lines" in their stories of behavioral problems that complicated medication administration schedules. Adult children frequently used humor to describe their role reversal with aged parents and the parents' forgetfulness, incontinence, or inability to dress without assistance. These accounts were placed in a taxonomy of humor response patterns that included categories for cognitive, affective, and behavioral responses. Second-level analysis created subcategories to reflect the function (relief or coping) that the humor served in the interview situation. It appears that humor is a useful communication tool for family caregivers that releases nervous energy about the interview process and the recall of difficult caregiving events. It is the responsibility of the interviewer to recognize the problems and issues embedded in the interview data and follow up humorous anecdotes with appropriate probes for additional information. Based on the results, a meta-humorous interaction theory is offered as an extension of the arousal-relief theory of humor. PMID- 11063287 TI - Lay theories of successful aging after the death of a spouse: a network text analysis of bereavement advice. AB - Social theories of successful aging attempt to explain how individuals adapt to changes characteristically associated with aging and to predict whether older adults' adaptations will lead to successful aging. The death of a spouse and the accompanying bereavement process entail dramatic changes to personal networks and experience to which individuals must adapt to age successfully. Network text analysis (including word frequencies, cluster analysis, and multidimensional scaling) of advice for adjusting to, and coping with, the loss of a spouse given by a sample of 60 bereaved spouses (mean age = 68) at 6 points in time after the death of their marital partner (3-4 weeks to 24 months) reveal respondents' lay theories of successful aging. Thematic clusters address social positioning and qualifiers, activity, communication, time, and spousal characteristics. Results indicate respondents frame their advice as unique to their context of social relationships while providing support for activity theory and negatively addressing disengagement theory. PMID- 11063288 TI - Comparison of serum insulin-like growth factor-1 and growth hormone levels in osteoporotic and non-osteoporotic postmenopausal women. AB - The purpose of this study was to compare the insulin-like growth factor-1 (IGF-1) levels and the growth hormone (GH) levels in osteoporotic and non-osteoporotic postmenopausal women. Eleven non-osteoporotic postmenopausal women and 9 women with untreated postmenopausal osteoporosis were included in the study. Bone mineral density (BMD) was assessed by dual energy X-ray absorbtiometry. Serum was assayed for calcium, phosphorus, alkaline phosphatase, bone-specific alkaline phosphatase, parathyroid hormone, IGF-1 and GH levels. IGF-1 levels were 98.8 +/- 43.5 ng/ml for osteoporotic women and 169.8 +/- 50.3 ng/ml for the women with normal BMD (p < 0.05). GH levels were 1.3 +/- 1.1 ng/ml and 1.3 +/- 1.0 ng/ml, respectively. When compared with normal postmenopausal women, IGF-1 levels were found to be lower in women with osteoporosis. IGF-1 seems to play an important role in the development of low bone mass and the present results suggest that IGF 1 is a useful predictor of the presence of osteoporosis. PMID- 11063289 TI - Cardiac involvement in patients with Wegener's granulomatosis. AB - Wegener's granulomatosis (WG) is a systemic necrotizing vasculitis, which could potentially affect any organ system. However, there have only been a few reports on cardiac involvement. We described the echocardiographic findings in nine patients affected by WG. A complete M-mode, two-dimensional, Doppler and color Doppler transthoracic echocardiogram was performed in nine patients (seven females and two males) affected by WG. In each patient, cardiac abnormality, for example, valvular damage, left ventricular global systolic dysfunction, or pericardial effusion, was detected. In particular, heart valve disease was found in eight patients, and in three cases, aortic valve insufficiency, which was severe enough to require surgical valve replacement, was observed. Cardiac involvement in patients with WG is common. In particular, there is a high frequency of aortic valve abnormalities. Thus, an echocardiographic study should be routinely performed. PMID- 11063290 TI - Extra-articular manifestations in 587 Italian patients with rheumatoid arthritis. AB - The aim of the study was to evaluate the frequency of extra-articular manifestations (EAMs) of rheumatoid arthritis (RA) in a series of patients from nine Italian rheumatology clinics. A total of 587 patients underwent direct questioning, complete physical evaluation, and review of medical records and laboratory data. The relationships between EAMs and the eosinophilic count, IgM rheumatoid factor (RF), and antinuclear antibodies (ANA) were studied. EAMs were present in 240/587 (40.9%) patients. The most common features were sicca syndrome (17.5%) and rheumatoid nodules (16.7%). EAMs were significantly more frequent in male patients (OR = 1.68), patients with ANA positivity (OR = 2.82), high anatomical class (OR = 2.3), and rheumatoid factor seropositivity (OR = 2.22). EAMs were more common in patients from southern Italy than in those from northern Italy (P < 0.001). EAMs seem to be rarer in Italy than in the Anglo-Saxon populations of northern Europe and the USA. Differences in prevalence of EAMs can exist even within the same country. PMID- 11063291 TI - Failure to detect Bartonella henselae infection in synovial fluid from sufferers of chronic arthritis. AB - Bartonella henselae causes granulomatous and indolent infection in the immune competent human, and angioproliferation in the context of persistent infection and impaired immunity. This bacterium is found in up to 40% of household cats, from which humans acquire it by either a cat scratch or a bite (hence the name, cat-scratch disease). Approximately 5% of Australian and US blood donors have serological evidence of past infection, but most associated illnesses are mild or subclinical. A number of lines of evidence prompted us to consider a relationship between rheumatoid arthritis (RA) and Bartonella infection. These include epidemiological associations with household pet exposure; apparent responsiveness of some RA cases to tetracycline therapy; the granulomatous and angioproliferative nature of Bartonella lesions; the insidiousness and high seroprevalence of this infection in the community; and even reported Bartonella infection mimicking juvenile RA. In a small group of patients with chronic arthritides, we found no direct evidence of humoral antibodies to, nor of persistent infection with, Bartonella henselae in synovial fluid. While larger and more invasive studies are likely to provide more confident exclusions of this hypothesis, this suggests that persistent Bartonella infection is unlikely to play a major role in RA. PMID- 11063292 TI - KL-6 as a novel marker for activities of interstitial pneumonia in connective tissue diseases. AB - The objective of this study was to determine the role of serum KL-6 levels as a marker for the activity of interstitial pneumonia in patients with connective tissue diseases. The serum concentrations of KL-6, a glycoprotein produced mainly by pulmonary type II epithelial cells, were measured in 21 patients with connective tissue disease. The activity of interstitial pneumonia was compared with the associated serum KL-6 concentrations. Serum KL-6 concentrations in patients with interstitial pneumonia were significantly higher than those in the controls. Among patients with active interstitial pneumonia, serum KL-6 concentrations following the treatment (after improvement) were significantly lower than the pretreatment values. The extent of the pulmonary fibrosis correlated positively with the serum KL-6 concentrations during the inactive phase of the interstitial pneumonia. These results suggest that sequential measurement of serum KL-6 levels is a new and useful means for the evaluation of interstitial pneumonia in patients with connective tissue diseases. PMID- 11063293 TI - Mediastinal mass and brachial plexopathy caused by subclavian arterial aneurysm in Behcet's disease. AB - Vascular involvement in Behcet's disease is divided into venous and arterial thrombosis and arterial aneurysmal formation. Subclavian arterial aneurysm rarely occurs in Behcet's disease; however, when it does occur, it causes serious aneurysmal rupture and local complications such as nerve compression and arterial ischemia. We describe the case of a 39-year-old male who presented with neurologic symptoms and signs of brachial plexopathy and mediastinal mass caused by Behcet's subclavian arterial aneurysm. This case shows that the occurrence of brachial plexopathy should be considered a manifestation of Behcet's disease, and that Behcet's aneurysm should be considered in the differential diagnosis of upper mediastinal mass. PMID- 11063294 TI - Relapsing polychondritis in childhood--case report and short review. AB - Relapsing polychondritis (RP) is a disease of unknown etiology and it is characterized by inflammation of the cartilage. While the clinical picture of RP in adults is well described, RP in childhood is poorly documented. We describe a young girl presenting with acute dyspnea, stridor and polyarthritis. The diagnosis of RP was made 2 years after first presentation, when auricular chondritis occurred. Based on a MEDLINE search, reports on RP in childhood were reviewed. The frequency of chondritis and systemic manifestations of RP in children was compared to data in adults and found to be very similar. RP in childhood can be a life-threatening and debilitating disease. PMID- 11063295 TI - An unusual case of systemic lupus erythematosus, lupus nephritis, and transient monoclonal gammopathy. AB - A 23-year-old female patient suffering from active systemic lupus erythematosus (SLE) was treated with azathioprine (2 mg/kg per day) and prednisone. Lupus nephritis class III with increasing proteinuria developed 28 months after disease onset. Treatment was switched to monthly pulse cyclophosphamide administered intravenously for 6 months (total dose 6.3 g), followed by oral azathioprine and low-dose prednisone to maintain partial remission. Eight months later, the patient developed an acute exacerbation of SLE with fever, proteinuria of 9.1 g/day, pancytopenia, and cerebral involvement with cephalgias and a grand mal seizure. She responded well to high-dose corticosteroids (500 mg prednisolone pulses over 3 days, i.v.) and was azathioprine switched from to methotrexate (12.5-15 mg per week). Under this treatment, lupus activity gradually decreased and the patient felt well again. Five years after the initial diagnosis of SLE, a rapidly increasing immunoglobulin G-kappa type (IgG-kappa) monoclonal gammopathy developed, reaching a maximal serum paraprotein concentration of 73.5 g/l. Bone marrow biopsy revealed 15% of moderately abnormal, highly differentiated plasma cells arranged in small clusters and expressing IgG-kappa. No bony lesions were detectable on skeletal radiographs. Pulses of dexamethasone (40 mg) were administered and led to a transient decrease of paraproteinemia to a minimum of 31.9 g/l, followed by an increase to 62 g/l. At that point, high-dose chemotherapy supported by autologous stem cell transplantation was considered. Due to an intermittent pneumococcal septicemia, methotrexate was discontinued and dexamethasone was replaced by 5-10 mg cloprednol. At this point, totally unexpectedly, the paraprotein decreased spontaneously without any further cytostatic treatment and was no longer detectable 1 year later. Concomitantly, plasma cell counts in bone marrow biopsies fell to below 5%. As SLE remained inactive, the patient became pregnant and gave birth to a healthy child. During late pregnancy, SLE activity flared up with rising proteinuria and blood pressure. Therefore, after delivery, cyclophosphamide (100 mg/day, orally) was readministered for 4 months, resulting in an improvement of kidney function with stable proteinuria of 1-2 g/l to date. Paraproteins are no longer detectable. In conclusion, this case report documents the rare event of transient paraproteinemia in a patient with SLE. A self-limiting regulatory defect in the control of a terminally differentiated B-cell clone may be the origin of this phenomenon. PMID- 11063296 TI - Performance and carcass merit of growing beef steers with chlortetracycline modified sensitivity to pituitary releasing hormones and fed two dietary protein levels. AB - This paper reports the effects of reduced sensitivity to growth hormone-releasing hormone and thyrotropin-releasing hormone through feeding a subtherapeutic level of chlortetracycline (CTC; 350 mg CTC/d) and two levels of dietary CP (10% and 13% of diet DM) on growth performance and carcass merit characteristics. Thirty two steers (initial average BW, 286 kg) were adapted to a common 13% CP diet consisting primarily of grass hay, corn, and soybean meal fed to gain 1.25 kg/d. The steers were assigned to four treatments (with or without CTC and 10% or 13% dietary CP in a factorial arrangement) and fed ad libitum amounts of diet for 91 d. Feed intake was determined daily and steers were weighed weekly. Steers were killed at the end of the feeding period for carcass merit determinations. Efficiency of BW gain was greater (P < .05) for steers fed the 13% CP diet than for the 10% CP diet and tended to be less for CTC-steers when the 10% CP diet was fed and greater for the CTC-steers when the 13% CP diet was fed (CTC x dietary CP interaction, P < .10). Feeding CTC increased (P < .01) fat over the longissimus muscle and marbling. This study is interpreted to indicate that the sustained effect of subtherapeutic feeding of CTC to cattle appears to increase fat deposition consistent with a reduced growth hormone and thyroid status reported earlier for these same steers. This would tend to increase energy utilization but may not necessarily produce a measurable increase in BW gain. PMID- 11063297 TI - Milk production and composition in captive Iberian red deer (Cervus elaphus hispanicus): effect of birth date. AB - This study describes milk production and milk composition of Iberian red deer (Cervus elaphus hispanicus) females (hinds) and the effect of calving date and BW change of hinds by milking over 34 wk. All hinds produced milk throughout the 34 wk study period, well over the standard lactation period. Total milk yield was 224.1 +/- 21.1 L, and daily production was 0.91 +/- 0.06 L. Milk yield decreased with lactation stage (P = 0.01) and the later a calf was born (P = 0.008), and it was greater in posterior quarters (P < 0.05). Milk yield was unaffected (P > 0.10) by side position or milking order of the udder. Milk production did not correlate with hind BW (P > 0.1). Hinds lost 4.4% BW during lactation (P < 0.001); losses increased the later a calf was born (P = 0.012). Iberian red deer milk had 11.5% fat, 7.6% protein, 5.9% lactose, and 26.7% DM. Stage of lactation affected fat (P < 0.001), protein (P = 0.002), DM (P < 0.001), and protein:fat ratio (P < 0.001), but not lactose (P > 0.1). These constituents became concentrated as lactation proceeded, and protein was substituted by fat. Calving date had a similar concentrating effect on fat (P < 0.001) and DM (P < 0.001), whereas it decreased lactose (P = 0.015) and protein (P = 0.002), thus producing a substitution of protein by fat (ratio of protein to fat, P < 0.001). Milking order of quarters or their position had no effect on milk composition (P > 0.10). Results suggest that milk production and milk energetic quality might increase by advancing calving date in red deer hinds. PMID- 11063298 TI - The effects of active immunization against gnRH on testicular development, feedlot performance, and carcass characteristics of beef bulls. AB - The objective was to determine the effects of a recombinant fusion protein anti GnRH vaccine on testicular development, feedlot performance, and carcass quality of beef bulls. Crossbred beef bulls (n = 58, average weight 306 kg, 9 mo of age), were randomly allocated to two groups and received either an anti-GnRH vaccine (GnRH) or placebo (Control) by intramuscular injection on d 0, 56, and 112. There were group effects (P < 0.01; as a percentage of Control) on testicular weight (53%), daily sperm production (40%), and epididymal sperm reserves (16%). There were group x time interactions (P < 0.0001) for scrotal circumference and serum testosterone concentrations; at slaughter, bulls in the GnRH group had a smaller (P < 0.05) scrotal circumference (28.3 vs 33.9 cm) and lower (P < 0.05) serum testosterone concentrations (2.2 vs 8.6 ng/mL) than those in the Control group. Average daily gain, feed intake, and feed efficiency were not different between treatments during the backgrounding phase (d 0 to 84). During the finishing phase (d 98 to 182), ADG was greater (P < 0.05) for bulls in the Control group (1.69 vs 1.42 kg/d), as was carcass weight (6.9%; P < 0.01). However, GnRH bulls had numerically better feed efficiency (6.12 vs 7.08 kg DMI/kg gain; P < 0.23) and shear force values for ribeye that were 16% lower (P < 0.14) than Control bulls, warranting further investigation. Vaccinating bulls against GnRH suppressed testicular function, with growth and carcass characteristics similar to that expected with steers. PMID- 11063299 TI - Feeder location did not affect performance of weanling pigs in large groups. AB - Crossbred pigs weaned at 17 d of age (n = 1,760; mean initial BW = 5.6 +/- 0.7 kg) were used in two 4-wk trials (four replicates per trial) to evaluate the effects of three pen designs on pig performance. The designs were 1) large group size (100 pigs/pen) with five two-sided feeders in a single, central location in the pen; 2) large group size (100 pigs/pen) with five two-sided feeders in multiple (five) locations in the pen; and 3) small group size (20 pigs/pen) with a single two-sided feeder in a central location in the pen. Each feeder provided two 20.3-cm-wide feeding places on each side. Pigs had free access to feed and water. Feeder-trough space (4 cm/pig) and floor-area allowance (0.17 m2/pig) were the same for all treatments. Pigs in the large-group treatments were lighter (15.6 and 15.6 vs 16.0 kg; P < 0.01) at the end of wk 4 and had lower ADG (358 and 357 vs 373 g; P < 0.01) and ADFI (510 and 521 vs 544 g; P < 0.01) during wk 2 through 4 than pigs in small groups. Gain:feed ratio was similar (P > 0.05) for all treatment groups throughout the study. For large groups, feed disappearance from each of the five feeders was similar (P > 0.05) for both multiple- and single-location treatments. In summary, large group size reduced pig growth performance, but the approach to providing multiple feeding locations that was employed in this experiment did not increase feed intake or growth performance of pigs in the large groups. PMID- 11063300 TI - Timing of realimentation of mature cows that were feed-restricted during pregnancy influences calf birth weights and growth rates. AB - The objective of this study was to determine the effect of feeding strategies in cows that allowed BW loss followed by BW gain on the efficiency of feed utilization for calf production. The first treatment (H-H-H) was designed to maintain body condition score of mature cows at 5.5 from the second trimester until the subsequent breeding season. The second treatment (L-H-H) was designed such that cows lost body condition during the second trimester and regained it during the third trimester and were equal in weight and body condition scores at parturition to cows assigned to the H-H-H treatment. The third treatment (L-L-H) was designed such that cows lost body condition during the second trimester and gained body condition after 28 d of lactation so that they would be equal to the other two treatments at breeding. Forty-eight cows were assigned to each treatment. Total DMI over the entire study did not differ between the H-H-H and L H-H treatments (P = 0.23), but intake on both were higher than the L-L-H treatment (P < 0.001). Calf birth weight of the H-H-H treatment did not differ (P = 0.43) from those of L-H-H, but both groups were greater than those of the L-L-H (P < or = 0.002) treatment. At 28 d of age, H-H-H (P = 0.008) and L-H-H (P = 0.007) calves weighed more than the L-L-H calves, but at 58 d of age there was no difference in calf BW among the treatments (P = 0.81). The percentage of cows that were diagnosed pregnant at weaning with their next calf did not differ (P = 0.71) among treatments. We interpret the results of this study to suggest that weight cycling in mature beef cows may be a viable management tool for decreasing food costs. PMID- 11063301 TI - Deletion of supplemental minerals and vitamins during the late finishing period does not affect pig weight gain and feed intake. AB - Two studies were conducted to determine the effects of eliminating the supplemental vitamin and minerals during the last 30 d of finishing on pig performance. The first study was conducted in a controlled university environment and the second under field conditions. Deletion of vitamin and mineral supplements for the last 30 d of finishing had no effects on pig performance or general health of pigs in either the university or the field trial. Besides lowering feed costs, deletion of dietary supplements may reduce the environmental impact of late finishing diets. PMID- 11063302 TI - Effects of supplemental zinc concentration and source on performance, carcass characteristics, and serum values in finishing beef steers. AB - Three studies were conducted to examine the effects of zinc concentration or source in diets of finishing beef steers. In Exp. 1, 108 (British x Continental) beef steers were supplemented with concentrations of added zinc (as ZnSO4) at 20, 100, or 200 mg/kg of dietary DM. No differences (P > 0.10) were noted among treatments for ADG or gain:feed for the 112-d finishing period. However, a linear (P < 0.10) decrease was noted in daily DMI with increasing zinc concentrations for the overall finishing period. No differences (P > 0.10) were noted in hot carcass weight; dressing percentage; longissimus muscle area; percentage of kidney, pelvic, and heart fat; or marbling score. There were, however, quadratic increases in s.c. fat thickness (P < 0.05) and yield grade (P < 0.01) with added zinc. In Exp. 2, 12 beef steers were used to examine effects of added dietary zinc on serum concentrations of cholesterol and fatty acid profiles. No differences (P > 0.10) were observed in cholesterol or fatty acids among the supplemental zinc levels. In Exp. 3, 84 Brangus- and Angus-sired steers were fed a steam-flaked corn-based diet containing 30 mg of supplemental zinc per kilogram of dietary DM from one of the following sources: 1) ZnSO4, 2) Zn amino acid complex, or 3) a zinc polysaccharide complex. No differences (P > 0.10) were noted for the overall 126-d trial for ADG, DMI, or gain:feed ratio. Percentage kidney, pelvic, and heart fat was increased (P < 0.10) in steers supplemented with ZnSO4 vs the average of Zn amino acid and Zn polysaccharide complexes. However, s.c. fat thickness was greater (P < 0.10) in steers supplemented with Zn amino acid and Zn polysaccharide complexes vs ZnSO4. Serum zinc concentration did not differ (P > 0.10) among zinc sources. Supplemental zinc concentration in finishing diets did not seem to influence feedlot performance and had a minimal impact on carcass quality. Either the organic or inorganic source can be included in finishing diets without affecting feedlot performance. PMID- 11063303 TI - Modeling selection for production traits under constant infection pressure. AB - This article presents a model describing the relationship between level of disease resistance and production under constant infection pressure. The model assumes that given a certain infection pressure, there is a threshold for resistance below which animals will stop producing, and that there is also a threshold for resistance above which animals produce at production potential. In between both thresholds animals will show a decrease in production, the size of decrease depending on the severity of infection and the level of resistance. The dynamic relationship between production and resistance when level of resistance changes, such as due to infection, is modeled both stochastically and deterministically. Selection started in a population with very poor level of resistance introduced in an environment with constant infection pressure. Mass selection on observed production was applied, which resulted in a nonlinear selection response for all three traits considered. When resistance is poor, selection for observed production results in increased level of resistance. With increasing level of resistance, selection response shifts to production potential and eventually selection for observed production is equivalent to selection for production potential. The rate at which resistance is improved depends on its heritability, the difference between both thresholds, and selection intensity. The model also revealed that when a zero correlation between resistance and production potential is assumed, the phenotypic correlation between resistance and observed production level increases for low levels of resistance and subsequently asymptotes to zero, whereas the phenotypic correlation between production potential and observed production asymptotes to 1.0. For most breeding schemes investigated, the deterministic model performed well in relation to the stochastic simulation results. Experimental results reported in literature support the model predictions. PMID- 11063304 TI - Relationships between human-animal interactions and productivity of commercial dairy cows. AB - This study examined the relationships between a number of stockperson and cow variables at 66 commercial dairy farms. Variables such as the attitudes and behavior of stockpeople toward their cows and the behavioral response to humans and productivity of cows were studied over one lactation. There were consistent and significant correlations between some of these stockperson and cow variables. For example, a positive attitude by stockpeople toward the behavior of dairy cows was negatively correlated with the number of forceful, negative, tactile interactions used by stockpeople in handling cows (r = -0.27, df = 127, P < 0.01). Furthermore, based on farm averages, the number of forceful, negative, tactile interactions used by stockpeople was negatively correlated with the percentage of cows approaching within 1 m of an experimenter in a standard test (r = -0.27, df= 64, P< 0.05). Although not confirming a fear-productivity relationship, a moderate but nonsignificant correlation was found between flight distance of cows to an experimenter in a standard test and milk yield (r = -0.27, df = 33, P > 0.05). Support for the existence of a negative fear-productivity relationship was the finding that the use of negative interactions by stockpeople was significantly and negatively correlated with milk yield, protein, and fat at the farm (r = -0.36, -0.35 and -0.33, respectively, df = 64, P < 0.01) and was significantly and positively correlated with milk cortisol concentrations at the farm (r = 0.34, df= 64, P < 0.01). Furthermore, the percentage of cows approaching within 3 m of an experimenter in a standard test was positively correlated with conception rate to the first insemination (r = 0.38, df = 46, P < 0.01). The significant correlations found in the present study between stockperson attitudes and behavior and cow behavior and productivity, although not evidence of causal relationships, indicate the possibility of targeting these human characteristics to reduce fear responses of dairy cows to humans and improve the cows' productivity. PMID- 11063305 TI - Farrowing hut design and sow genotype (Camborough-15 vs 25% Meishan) effects on outdoor sow and litter productivity. AB - Performance measures were evaluated for 125 outdoor sows and litters of two crossbred genotypes (Camborough-15 and 25% Meishan) and in two farrowing hut designs (American-style and English-style hut). Contemporary breeding groups of second-parity sows were evaluated in an intensive, outdoor research unit. Sow genotype and hut designs were arranged factorially. Seven complete blocks were evaluated over a 21-wk period. No interactions between environment and genotype were identified for sow and litter productivity. Litters farrowing in the English style huts weaned 1.5 more (P < .05) piglets per sow (because of a lower preweaning mortality, P = .05) than did litters in the American-style huts. The 25% Meishan weaned 1.7 more (P < .01) pigs per sow than Camborough-15, because of a greater number of piglets born alive. The effects of hut style and genotype were additive and 25% Meishan sows in English-style huts weaned an average (+/- SEM) of 11.1 +/- .83 piglets per sow. The English-style arc hut design may improve outdoor pig production and increase competitiveness of the intensive, outdoor system. The 25% Meishan genotype has potential for increased pigs weaned per litter that must be considered in light of other features of this genotype such as body composition. PMID- 11063306 TI - Effect of the callipyge gene on muscle growth, calpastatin activity, and tenderness of three muscles across the growth curve. AB - Changes in muscle growth, calpastatin activity, and tenderness of three muscles were assessed in 20 callipyge and 20 normal wether lambs slaughtered at live weights (LW) of 7, 20, 36, 52, and 69 kg. At 24 h postmortem, the longissimus (LM), semimembranosus (SM), and supraspinatus (SS) muscles were removed and weighed and samples were obtained for calpastatin activity (CA; 24 h) and Warner Bratzler shear force (WBS; aged 6 d). For muscle weights and calpastatin activity, the weight group x muscle x phenotype interaction was significant (P < 0.05). Muscle weights were similar (P > 0.05) between phenotypes for all three muscles at 7 kg LW. At 20 kg LW, the LM and SM muscles from the callipyge lambs were heavier (P < 0.05) than those from normal lambs; however, the SS did not differ (P > 0.05) between phenotypes at 7, 20, or 52 kg. From 20 to 69 kg LW, the LM and SM weights were 42 and 49% heavier (P < 0.05) for callipyge than for normal lambs. Calpastatin activity of the callipyge LM was greater (P < 0.05) than that of normal LM at 36, 52, and 69 kg. In the callipyge LM, CA was similar (P > 0.05) at 20, 36, and 52 kg LW and did not differ (P > 0.05) from 7-kg or 69 kg values. Calpastatin activity declined (P < 0.05) across the growth curve for the SM and SS, but values were higher (P < 0.05) in the SM in callipyge than in normal lambs. Shear force values of the LM were lower (P < 0.05) for normal lambs at 36, 52, and 69 kg LW than for callipyge lambs. In the SM and SS, WBS values decreased (P < 0.05) across the growth curve, but values were higher (P < 0.05) for callipyge lambs in the SM only. These data indicate that the selective muscular hypertrophy of the callipyge phenotype develops during the postnatal growth period between 7 and 20 kg LW (19 and 100 d of age). Longissimus and semimembranosus muscles in the callipyge lambs were over 40% heavier from 20 to 69 kg LW; however, they also had higher levels of calpastatin activity and Warner Bratzler shear force during this time period, indicating the need for postmortem tenderization treatments to improve palatability. PMID- 11063307 TI - Rigor temperature and meat quality characteristics of lamb longissimus muscle. AB - The present experiment was conducted to determine the effect of muscle temperature during the prerigor and early postrigor period on meat tenderness, postmortem proteolysis, calpain system activity, water-holding capacity, and color. Lamb longissimus muscle (n = 14) from the right and left carcass sides was excised immediately after dressing, divided into an anterior and posterior sample, vacuum-packaged, and stored overnight at 5 to 35 degrees C. Further storage, up to 14 d postmortem, was at 2 degrees C. Tenderness at 1 d postmortem, tenderization during further storage, and postmortem proteolysis were negatively affected by overnight incubation above 25 degrees C. This effect could be explained by an effect of temperature on muscle contraction and activity of the calpain system. Muscle contraction was at a minimum after incubation at 15 degrees C. Water-holding capacity was negatively affected by incubation above 25 degrees C. Color scores improved with increasing incubation temperature at 1 d postmortem. However, after 14 d of postmortem storage, no differences in color scores were observed. Based on the present results and results of other groups, a temperature around 15 degrees C at the onset of rigor seems optimal to maximize tenderness without having detrimental effects on water-holding capacity or color. PMID- 11063308 TI - Fatty acid composition, including conjugated linoleic acid, of intramuscular fat from steers offered grazed grass, grass silage, or concentrate-based diets. AB - The effects of grazed grass, grass silage, or concentrates on fatty acid composition and conjugated linoleic acid (cis-9, trans-11-18:2; CLA) concentrations of i.m. fat of steers fed to achieve similar carcass growth rates were investigated. Fifty steers were divided into 10 blocks based on body weight and assigned at random from within blocks to one of five dietary treatments. The experimental rations offered daily for 85 d preceding slaughter were 1) grass silage for ad libitum intake plus 4 kg of concentrate, 2) 8 kg of concentrate plus 1 kg of hay, 3) 6 kg of grazed grass DM plus 5 kg of concentrate, 4) 12 kg of grazed grass DM plus 2.5 kg concentrate, or 5) 22 kg of grazed grass DM. The concentration of polyunsaturated fatty acids (PUFA) in i.m. fat was higher (P < .05) for steers offered ration 5 than for those given any other ration. Decreasing the proportion of concentrate in the diet, which effectively increased grass intake, caused a linear decrease in the concentration of i.m. saturated fatty acids (SFA) (P < .01) and in the n-6:n-3 PUFA ratio (P < .001) and a linear increase in the PUFA:SFA ratio (P < .01) and the conjugated linoleic acid concentration (P < .001). The data indicate that i.m. fatty acid composition of beef can be improved from a human health perspective by inclusion of grass in the diet. PMID- 11063309 TI - Relationship of beef longissimus tenderness classes to tenderness of gluteus medius, semimembranosus, and biceps femoris. AB - The objective of this study was to determine the relationship of longissimus tenderness classes to tenderness of three other major muscles. Ninety-eight crossbred steers and heifers (14 to 17 mo of age) were humanely slaughtered over 9 wk and the carcasses were chilled 48 h at 0 degrees C. At 48 h postmortem, carcasses were assigned to one of three tenderness classes (tender < or = 26 kg, intermediate = 26 to 42 kg, tough > or = 42 kg) using slice shear force from the MARC Beef Classification System (n = 20, 67, and 11, respectively). The longissimus thoracis, gluteus medius, semimembranosus, and biceps femoris were removed, aged at 2 degrees C, and frozen at -30 degrees C at 14 d postmortem. Two 2.54-cm-thick steaks were obtained from each muscle, thawed to 5 degrees C, cooked with a belt grill at 163 degrees C for 5.5 min, and served warm to an eight-member trained descriptive attribute panel. Panelists evaluated each sample for tenderness, connective tissue amount, juiciness, and beef flavor intensity on 8-point scales. The mean 2-d longissimus slice shear force values were 20.7, 34.4, and 46.3 kg, respectively, for the "tender," "intermediate," and "tough" classes. Tenderness ratings were lowest (P < 0.05) for the "tough" class and highest (P < 0.05) for the "tender" class for all muscles except the gluteus medius, for which the "tender" and "intermediate" classes were not different (P > 0.05; longissimus, 7.7, 7.1, 6.3, and 7.1; semimembranosus, 6.4, 5.8, 5.1, and 5.8; biceps femoris, 5.9, 5.4, 4.8, and 5.4; gluteus medius, 6.8, 6.5, 5.8, and 6.5 for the "tender," "intermediate," "tough," and "unsorted" classes, respectively). The magnitude of the differences in tenderness ratings between the "tender" and "intermediate" classes and between the "intermediate" and "tough" classes was similar for all muscles. The percentages of tenderness ratings greater than 5.0 (slightly tender) for the "tender" and "unsorted" classes, respectively, were as follows: longissimus, 100 and 95%; semimembranosus, 95 and 85%; gluteus medius, 100 and 94%; and biceps femoris, 95 and 81%. The simple correlations between longissimus and the other muscles for tenderness ratings were as follows: semimembranosus, 0.58; biceps femoris, 0.43; and gluteus medius, 0.68. These data indicate that early-postmortem longissimus slice shear force could be used to classify top sirloin, top round, and bottom round cuts for tenderness. PMID- 11063310 TI - The effect of the Halothane and Rendement Napole genes on carcass and meat quality characteristics of pigs. AB - The objective of this study was to determine the effects of the Halothane (N) and Rendement Napole (RN) genes on carcass and meat quality characteristics in pigs. Halothane and RN carrier (Nn/RN- rn+) Hampshire boars (n = 4) were mated to dams that were homozygous for the normal allele of both genes (NN/rn+ rn+) to produce progeny of four genotypes: 1, NN/rn+ rn+ (n = 31); 2, Nn/rn+ rn+ (n = 27); 3, NN/RN- rn+) (n = 30); and 4, Nn/RN- rn+ (n = 23). A DNA test was used to determine Halothane genotype, and longissimus glycolytic potential was used to predict the RN genotype. Pigs were reared under standard conditions to approximately 120 kg live weight and slaughtered at a commercial plant, and carcass characteristics and meat quality were evaluated. Halothane carriers (Nn/ _ _), in comparison to Halothane normal (NN/_ _) pigs, had shorter carcasses, lower longissimus ultimate pH, higher Minolta L* and b* values, and greater drip loss. Rendement Napole gene carriers (_ _/RN- rn+) had higher L* and b* values and drip and cooking loss and lower longissimus ultimate pH than homozygous recessive animals (_ _/rn+ rn+). There were Halothane x RN genotype interactions (P < 0.05) for subjective color, firmness, and marbling scores, and for shear force. Animals that were normal for both genes (NN/rn+ rn+) had the highest subjective scores for color (2.60, 1.88, 1.85, and 1.95, SE = 0.181, P < 0.05), firmness (2.53, 2.03, 2.10, and 1.89, SE = 0.182, P < 0 .05), and marbling (2.11, 1.44, 1.53, and 1.55, SE = 0.153, P < 0 .05) for genotypes 1, 2, 3, and 4, respectively, suggesting darker, firmer muscle with a higher level of marbling for this genotype. Shear force was highest for Nn/rn+ rn+ animals (3.83, 4.41, 3.79, and 3.70, respectively, SE = 0.172, P < 0.05). Gilts had less s.c. backfat thickness, greater longissimus muscle area, and lower subjective marbling scores than barrows. There was no effect (P > 0.05) of gender on other meat quality traits. This study illustrates the negative effects of the Halothane and RN genes on fresh pork quality and suggests that in combination the detrimental effects of the two genes are additive for ultimate pH, objective color, and water-holding capacity. PMID- 11063311 TI - Comparison of phytase from genetically engineered Aspergillus and canola in weanling pig diets. AB - Ninety-six crossbred pigs with an average weight of 9.0 kg were used in a 5-wk trial to compare the efficacy of genetically engineered Aspergillus ficuum phytase, expressed in Aspergillus niger (Natuphos) or in canola seed (Phytaseed), for enhancing the utilization of phytate P in corn-soybean meal-based diets fed to young pigs and to evaluate the safety of Phytaseed phytase. Three levels of the two sources of phytase (250, 500, or 2,500 U/kg of diet) were added to a corn soybean meal basal diet containing .35% total P, .09% available P, and .50% Ca. There were six pens per treatment (one barrow and one gilt/pen), except that the diet without added phytase was fed to 12 pens of pigs. Pen feed consumption and BW were recorded weekly. During wk 5, pen fecal samples were collected for determination of apparent digestibilities of DM, Ca, and P. At the end of wk 5, all barrows were killed, and the 10th rib on both sides was removed for determination of shear force and energy. Thirty pigs (six from the diet without added phytase and the diets with 500 and 2,500 U/kg phytase from both sources) were randomly selected for gross necropsy and histologic evaluation of liver, kidney, and bone tissues. Both sources of phytase were equally effective in increasing (P < .05) daily gain, gain:feed, apparent digestibilities of DM, P, and Ca, and 10th rib measurements. Fecal P excretion was reduced with phytase addition. Feed intake was increased by phytase levels during wk 4 to 5. No significant abnormalities were seen in any of the 30 pigs necropsied. The fit of a nonlinear function revealed that most measurements were reaching a plateau at 2,500 U/kg phytase. In summary, based on performance, bone measurements, and digestibilities of P, Ca, and DM of young pigs, the efficiency of Phytaseed was similar to that of Natuphos for enhancing the utilization of phytate P in corn soybean meal-based diets. General necropsy and histologic examination of tissues indicated no toxic effect of phytase. PMID- 11063312 TI - Effects of dietary valine concentration on lactational performance of sows nursing large litters. NCR-42 Committe on Swine Nutrition. AB - A cooperative study, using 231 primiand multiparous crossbred sows from six experiment stations (IN, KS, MI, MN, ND, and OH), was conducted to determine the effects of elevating dietary valine concentration in corn-soybean meal diets on lactational performance of sows nursing large litters. Crossbred sows were fed diets containing a minimum of .60% lysine during gestation. Sows were allotted at farrowing to four dietary valine concentrations, .80, .95, 1.10, and 1.25%. Crystalline L-valine replaced cornstarch to maintain a constant ratio of corn:soybean meal across diets. Dietary lysine, provided by corn, soybean meal, and .15% crystalline L-lysine x HCl, was .90% in all diets. Sows were allowed ad libitum access to feed. Sows were weighed within 24 h after farrowing, and all litters were adjusted to > or = 10 pigs/litter by d 2 following farrowing. Average sow parity, number of pigs on d 2, and lactation length for the four treatments were, respectively, 2.3, 2.3, 2.3, 2.5; 10.9, 10.8, 10.8, 10.7; and 25.1, 24.5, 25.2, 25.0 d. The ADFI during lactation was 5.87, 5.77, 5.87, and 5.74 kg (P > .50); hence, valine intakes were 41, 48, 55, and 61 g/d (linear, P < .01). Lysine intake ranged from 51.5 to 52.7 g/d (P > .50). Sow weight after farrowing averaged 198 kg (P > .60). Overall pig survival to weaning was high (>92%), and the number of pigs weaned (10.1, 10.3, 10.3, 10.3) did not differ (P > .30) among treatments. Litter weaning weights (73.6, 73.6, 74.5, 72.6 kg), litter weight gains (55.1, 55.1, 56.0, 54.1 kg), sow weight change during lactation (-4.9, -5.4, -4.8, -6.3 kg), and return-to-estrus interval (7.5, 6.4, 6.9, 8.2 d) were not affected (P > .30) by dietary valine. There were no station x treatment interactions (P > .50). These results indicate no benefit of elevated dietary valine for lactating sows nursing > or = 10 pigs and consuming a corn soybean meal diet containing .90% lysine and .80% valine. PMID- 11063313 TI - Effect of starter feeding program on growth performance and gains of body components from weaning to market weight in swine. AB - Three experiments were conducted to test the hypothesis that different starter feeding programs (High, high quality; Low, low quality) will affect growth performance and body composition of pigs from weaning to market weight and that this effect may be influenced by gender (barrows or gilts) and breed (F, Yorkshire-Duroc x Hampshire; P, PIC Camborough 15 x PIC line 405). In Exp. 1, 21 +/- 4-d-weaned F pigs (n = 90) were used in a 2 (High or Low) x 2 (barrows or gilts) factorial design. In Exp. 2, 21 +/- 3-d-weaned pigs (n = 184) were used in a 2 (F or P) x 2 (High or Low) x 2 (barrows or gilts) factorial design. In Exp. 3, 21 F pigs from each gender and feeding program treatment were killed at d 0, 3, 7, 14, 42, 82, or 152 postweaning for evaluating body composition. Two starter feeding programs (High or Low) were applied to pigs for 6 wk postweaning. Pigs from both High and Low treatments were provided the same corn-soybean meal-based diets for the growing and finishing periods. Although the ADG of all pigs receiving the High treatment during the early starter period were higher (P < .01) than those of the Low, the terminal BW of F barrows were similar between High and Low (Exp. 1 and 2) and those of gilts were similar between High and Low (Exp. 2) (P > .80). However, the BW of P pigs receiving the High treatment, regardless of gender, tended to be heavier than those receiving the Low (Exp. 2) and F barrows receiving the High treatment tended to be heavier than those receiving the Low (Exp. 1). For the first 7 d postweaning, the High-fed pigs gained more protein (P < .05) and lost less fat (P < .05) than Low-fed pigs. During the growing-finishing period, the Low-fed pigs exhibited compensatory protein gain and achieved a body protein content similar (P < .60) to High-fed pigs by termination. Protein gains from weaning to termination between High- and Low-fed pigs were not different in Exp. 2 and 3. The protein gain of gilts was higher (P < .05) than that of barrows. Similarly, fat gain within genders was not affected by starter feeding program. Fat gain of gilts, however, was lower (P < .08) than that of barrows in Exp. 3. In conclusion, the nutritional quality of the starter feeding program affected growth performance immediately after weaning but did not affect protein gain over the entire production period. PMID- 11063314 TI - Growth-promoting efficacy in young pigs of two sources of zinc oxide having either a high or a low bioavailability of zinc. AB - Commercial sources of zinc oxide (ZnO) differ widely in Zn relative bioavailability (RBV), but it is unknown whether growth-promoting efficacy in young pigs is influenced by RBV of the ZnO sources used. We compared a low-RBV (39%) ZnO manufactured by the Waelz process (W) to a high-RBV (93%) ZnO manufactured by the hydrosulfide process (HS). Antibacterial agents were included in the diet in only one of the four trials (Exp. 4). In Exp. 1, pigs (n = 36, 6.5 kg, 28 d of age) were randomly assigned in three replicates to receive 0, 1,500, or 3,000 mg Zn/kg from HS Zn in a 21-d growth assay. Growth rates and feed intake responded linearly (P < 0.01) to incremental doses of Zn. In Exp. 2, pigs (n = 60, 6.1 kg, 28 d of age) were randomly assigned in five replicates to receive either 0 or 1,500 mg W or HS Zn/kg during a 21-d feeding period. Growth performance was improved (P < 0.01) by the addition of ZnO. During wk 1, however, pigs receiving HS Zn grew faster (P < 0.03) than those receiving W Zn, but the difference diminished to a trend (P < 0.08) during wk 2. Morphology of duodenal, jejunal, and ileal intestinal sections was examined at d 21 of the assay, but neither source of ZnO had an effect on crypt depth or on villus height or width. In Exp. 3, weaned pigs (n = 48, 5.4 kg, 21 d of age) were randomly assigned in four replicates to the same dietary treatments as in Exp. 2 for a 17-d growth assay. Growth performance was improved (P < 0.05) by the addition of ZnO, but no difference was detected between the two sources. In Exp. 4, pigs (n = 60, 6.2 kg, 28 d of age) were randomly assigned in five replicates to receive either 0 or 1,500 mg/kg W or HS Zn in an 11-d growth assay wherein antimicrobial agents were included in the basal diet. Growth rates during the first 6-d were improved (P < 0.06) by the addition of ZnO, with a trend (P < 0.10) for greater weight gain in pigs receiving HS than in those fed W Zn. During the entire 11-d, however, there was no difference in growth rates between pigs fed the two sources of ZnO. In conclusion, RBV of Zn in ZnO did not substantially affect the growth-promoting efficacy of ZnO in young pigs fed diets with or without antimicrobial agents. PMID- 11063315 TI - Total radioactive residues and clenbuterol residues in swine after dietary administration of [14C]clenbuterol for seven days and preslaughter withdrawal periods of zero, three, or seven days. AB - Nine barrows (23.8 +/- 0.9 kg) and 9 gilts (23.1 +/- 0.9 kg) were used to determine the disposition of radiocarbon after oral [14C]clenbuterol (4-amino alpha-[t-butylaminomethyl]-3,5-dichlorobenzyl [7-(14)C]alcohol hydrochloride) administration and to determine total and parent residues in edible tissues. Three barrows and three gilts, housed in metabolism crates, were fed 1 ppm [14C]clenbuterol HCl for seven consecutive days in three separate trials; a single barrow and gilt from each trial was slaughtered after 0-, 3-, or 7-d preslaughter withdrawal periods. Urine and feces were collected during the dosing and the withdrawal period; edible and inedible tissues were collected at slaughter. Total recovery of radiocarbon was 94.2 +/- 6.5%. Total clenbuterol absorption was greater than 75% for barrows and 60% for gilts. Total radioactive residues in tissues were not different (P > 0.05) between barrows and gilts. Concentrations of parent clenbuterol in liver, kidney, skeletal muscle, adipose tissue, and lung did not differ between barrows and gilts (P > 0.05). Total radioactive and parent residues declined in tissues as withdrawal period increased. After the 0-d withdrawal period, total liver residues (286 ppb) were approximately equal to lung residues, twice those of the kidney, and about 15 times those of adipose tissue and skeletal muscle. After a 7-d withdrawal period, total radioactive residues in liver (15 ppb) were roughly three times greater than lung, kidney, and adipose tissue total residues and about 13 times those of skeletal muscle total residues. Parent clenbuterol represented 79, 63, 42, 67, and 100% of the total radioactive residue in adipose tissue, kidney, liver, lung, and skeletal muscle, respectively, in hogs slaughtered with a 0-d withdrawal period. With increasing withdrawal period, the percentage of total radioactive residue present as parent clenbuterol within edible tissues (including lung) decreased, so that after a 7-d withdrawal period, 7, 16, and 29% of the total residue was composed of parent clenbuterol in kidney, liver, and lung, respectively. After a 7-d withdrawal period, parent clenbuterol exceeded the European maximum residue limit (0.5 ppb) 4.6-fold in liver and 2.4-fold in lung. In muscle, clenbuterol was approximately 40 times the limit after a 0-d withdrawal period but had dropped below 0.5 ppb after a 3-d withdrawal period. Results from this study indicate that clenbuterol HCl is well absorbed in swine and that the use of clenbuterol in this species in an off-label manner is inconsistent with human food safety standards used in developed countries. PMID- 11063316 TI - Predicting growth in angus bulls: the use of GHRH challenge, insulin-like growth factor-I, and insulin-like growth factor binding proteins. AB - Plasma IGF-I, IGF binding protein-2 (IGFBP-2), and IGFBP-3 were quantified in growing Angus bulls (n = 56) to determine their relationship with postweaning growth and carcass ultrasound measurements. In addition, GH response to GHRH challenge (area-under-the-curve GH [AUC-GH) was determined for each bull as part of a previous study. Blood was collected by jugular venipuncture at the start of a 140-d postweaning growth performance test and at 28 d intervals for plasma IGF I determination by RIA. Plasma IGFBP-2 and -3 content was measured at the start of the study, on d 70, and d 140 by Western ligand blotting. Individual weights and hip heights were measured every 28 d during the study and carcass longissimus muscle area, intramuscular fat percentage, and carcass backfat were estimated by ultrasound on d 140. Greater plasma IGF-I at the start of the performance test was associated with reduced postweaning ADG and increased longissimus area. Throughout the performance test period, the correlations between plasma IGF-I and hip height were consistently positive, ranging from 0.10 to 0.38, but the correlations between ADG and IGF-I varied from -0.32 to 0.31. Age-adjusted d-1 plasma IGFBP-2 was related to ADG during the performance test, explaining nearly 30% of the variation in ADG. A model combining weaning age, IGFBP-2, and AUC-GH showed a strong relationship with ADG (R2 = 0.40). Plasma IGFBP-2 and -3 were not related to carcass characteristics, and IGFBP-3 was not related to growth rates. This study provides additional evidence for the variable relationship between plasma IGF-I and growth rates in cattle. A significant positive relationship between plasma IGFBP-2, AUC-GH, and postweaning ADG warrants further investigation. PMID- 11063317 TI - Production of insulin-like growth factor-I by granulosa cells but not thecal cells is hormonally responsive in cattle. AB - To determine whether the hormonal regulation of IGF-I production differs between granulosa and thecal cells in cattle, granulosa and thecal cells from bovine follicles were collected, cultured for 2 d in medium containing 10% fetal calf serum, washed, and then treated for an additional 24 h in serum-free medium with various hormones. In Exp. 1, granulosa cells were treated with 0 or 100 ng/mL of insulin and(or) 50 ng/mL of follicle-stimulating hormone (FSH), insulin plus 10 ng/mL of epidermal growth factor, or insulin plus 10 ng/mL of basic fibroblast growth factor. In Exp. 2, thecal cells were treated as described in Exp. 1 except that 100 ng/mL of luteinizing hormone (LH) was used instead of 50 ng/mL of FSH. In Exp. 3, granulosa and thecal cells were treated with 0 or 30 ng/mL of cortisol with or without 100 ng/mL of insulin, 300 pg/mL of glucagon, or glucagon plus insulin. In Exp. 4, granulosa and thecal cells were treated with 0 or 300 ng/mL of estradiol with or without 100 ng/mL of insulin and(or) 100 ng/mL of LH. At the end of treatment, medium was collected, concentrated with Centricon-3 concentrators, and assayed for IGF-I by radioimmunoassay. Cell numbers were determined by Coulter counting at the end of culture. Thecal cells produced low amounts of IGFI (0.48 +/- 0.04, 0.63 +/- 0.03, and 0.82 +/- 0.03 ng per 100,000 cells per 24 h in Exp. 2, 3, and 4, respectively), and this production was not influenced (P > 0.05) by the various treatments. In contrast, IGF-I production by granulosa cells (2.0 to 6.2 ng per 100,000 cells per 24 h) was influenced by treatment in Exp. 1, 3, and 4 and was greater than IGF-I production by thecal cells (Exp. 2, 3, and 4). Alone, insulin, FSH, LH, and cortisol (but not estradiol) each decreased (P < 0.05) granulosa-cell IGF-I production by 20 to 57%; combined treatments of insulin plus FSH or insulin plus cortisol decreased IGF-I production to levels seen with insulin alone. Glucagon had no effect (P > 0.10) on IGF-I production in the absence or presence of insulin. In the presence of insulin, epidermal growth factor, basic fibroblast growth factor, and estradiol decreased (P < 0.05) IGF-I production below that observed for insulin alone. These results indicate that, during follicular development in cattle, changes in intrafollicular levels of IGF-I may be due to hormonally-induced changes in granulosa-cell, but not thecal-cell, IGF-I production. PMID- 11063318 TI - Cellular uptake of valine by lactating porcine mammary tissue. AB - The cellular uptake of branched-chain amino acids in mammary tissue is important for understanding their role in milk synthesis in the sow. This study characterized the kinetic properties and substrate specificity of the valine uptake system in the porcine mammary gland. Mammary tissue was collected from lactating sows at slaughter and tissue explants were incubated in media containing isosmotic salt and amino acids of interest, plus [3H]valine tracer. Valine uptake was time-dependent and was dependent on the presence of sodium, as indicated by a reduction in uptake when sodium in the medium was replaced by choline. The valine transport system in porcine mammary tissue had a Km of 0.64 mM, a Vmax of 1.84 mmol-kg cell water(-1) 30 min(-l), and a Kd (diffusion constant) of 1.16 L x kg cell water(-1) x 30 min(-1). Valine uptake was inhibited by leucine and alpha-aminoisobutyric acid and by high concentrations of L alanine, L-lysine, cycloleucine, L-glutamine, and L-methionine, but not by 2 (methyl-amino)-isobutyric acid. This transport system is the primary system responsible for uptake of valine, and probably other branched-chain amino acids, in lactating sow mammary tissue. Physiological concentrations of valine in the blood are below the Km of the specific valine transport system and well below the diffusion uptake capabilities. The kinetic parameters of this valine transport system should not be limiting to valine uptake for milk protein synthesis. However, competition of valine uptake with branched-chain amino acids, as well as with other amino acids, may affect valine uptake in lactating tissue. PMID- 11063319 TI - Adaptations of glucose metabolism in multiparous sows: effects of pregnancy and feeding level. AB - This experiment was undertaken to determine whether pregnancy affects glucose metabolism and insulin sensitivity in sows fed at different levels. Four replicates of six multiparous Large White sows were involved. In each replicate, four sows were inseminated on the first postweaning estrus (pregnant group) and the two remaining were kept nonpregnant. Half of the sows of each group were fed 2.5 kg/d (low level) and the others 4 kg/d (high level) of the same standard pregnancy diet. Jugular catheters were implanted 2 to 3 d after estrus. Plasma concentrations of glucose, insulin, and FFA were determined before and during the 4 h following the morning meal at 10, 30, 59, 87, 93, 101, and 110 d of gestation and at equivalent periods for the nonpregnant sows. Intravenous glucose tolerance tests were performed at 33, 71, 85, 96, and 108 d by i.v. injection of 0.5 g glucose/kg BW. Compared with the glycemia before the meal, all the sows showed hyperglycemia 30 min after the initiation of the meal and hypoglycemia thereafter, with a minimum reached at approximately 75 min. Insulinemia increased from 20 min after food access, reached a maximum at 40 min, and returned to the basal level after 180 min. The higher feeding level increased plasma insulin and lowered plasma glucose levels. Glycemia and insulinemia profiles changed from 87 d onward in the pregnant sows. The peak of glucose induced by the meal was higher, and the subsequent period of hypoglycemia almost disappeared. The area under the insulin curve was unchanged, but insulin secretion was delayed. The glucose tolerance tests showed that between d 85 and 108 the half-life of injected glucose increased and insulin secretion was delayed in the pregnant sows. Compared to the following stages, plasma FFA were high before and after the meal at 10 d, which most likely was a residual effect from the previous gestation/lactation cycle. They were lower from 30 to 101 d in the pregnant and nonpregnant sows. At 110 d, fasting FFA were high again in the pregnant sows only, very likely in relation to the preparation for lactation. This experiment showed that insulin sensitivity decreases after 85 d of pregnancy in multiparous sows. PMID- 11063320 TI - Ability of induced corpora lutea to maintain pregnancy in beef cows. AB - Experiments were conducted in beef cows without a primary CL, in which pregnancy had been maintained with exogenous progestogen. In preliminary trials, replacement CL induced ipsilateral to the embryo and after, rather than before, d 36 of pregnancy, maintained more pregnancies after withdrawal of exogenous progestogen (13/13 vs 2/6; P < 0.05). In Exp. 1, in cows with replacement CL induced by treatment with hCG on d 28 of pregnancy, treatment with flunixin meglumine on d 31 through 37 did not increase maintenance of pregnancy. Experiment 2 was conducted to evaluate directly the effects of concentrations of PGF2alpha and estradiol-17beta during d 31 through 35 of pregnancy on maintenance of pregnancy by replacement CL induced between d 28 and 31. In cows that maintained pregnancy while progestogen was provided, maintenance of pregnancy after withdrawal of exogenous progestogen tended to be greater with high (5/5) than with low (2/6; P < 0.10) concentrations of PGF2alpha and greater with low (6/7) than with high (2/6; P = 0.10) concentrations of estradiol-17beta. Secretion of progesterone by replacement CL was greater (P < 0.05) in cows with high than in those with low concentrations of PGF2, during d 31 through 35. Prostaglandin F2alpha may facilitate attachment of the bovine embryo (d 30 to 40) in a manner similar to that reported for implantation in other species. Cows that did not form CL in response to hCG on d 28 to 31 responded well when retreated after d 36. Again, maintenance of pregnancy was greater when replacement CL were induced after (9/9) rather than before d 36 (8/16; P < 0.05). PMID- 11063321 TI - Chromium supplementation does not influence glucose metabolism or insulin action in response to cold exposure in mature sheep. AB - An isotope dilution method using [U-(13)C] glucose infusion and a glucose clamp approach were applied to determine the effects of supplemental Cr and cold exposure on blood glucose turnover rate and tissue responsiveness and sensitivity to insulin in eight sheep. The daily profiles of blood metabolites and hormones were also determined. The sheep consumed diets containing either 0 or 1 mg of Cr/kg from a high-Cr yeast and were exposed from a thermoneutral environment (20 degrees C) to a cold environment (0 to 4 degrees C) for 9 d. The experiment used a crossover design. Body weight was lost (P = 0.02) during cold exposure, regardless of Cr supplementation. Blood glucose turnover rate and the maximal glucose infusion rate did not differ between diets, but both were higher (P = 0.0004 and P = 0.0001, respectively) during cold exposure than in the thermoneutral environment. The plasma insulin concentration at half-maximal glucose infusion rate changed with neither diet nor environment. Plasma concentrations of glucose and NEFA increased (P < 0.05) during cold exposure for both diets. In sheep, Cr supplementation, 1 mg/kg of diet as high-Cr yeast, has little influence on blood glucose metabolism and insulin action, whereas cold exposure enhances both without further modification by Cr supplementation. PMID- 11063322 TI - Brown midrib-3 corn silage improves digestion but not performance of growing beef steers. AB - The brown midrib-3 (bm3) gene mutation has been incorporated into corn plants to potentially improve fiber digestibility. The objectives of this study were to determine the effect of bm3 corn silage on digestion and performance of growing beef steers and to determine whether limiting intake would further enhance fiber digestibility of bm3 corn silage. A bm3 hybrid and its isogeneic normal counterpart were harvested at three-quarters kernel milk line. Neutral detergent fiber, ADF, and ADL were 4.5, 6.9, and 1.9 units lower, respectively, and DM was 5.4 units higher for bm3 than for normal silage. In Trial 1, eight ruminally fistulated Angus crossbred steers (224 +/- 24 kg) were randomly assigned to a 2 x 2 factorial arrangement of treatments in a replicated 4 x 4 Latin square design. Steers had ad libitum feed access or were restricted to 80% of ad libitum intake of diets containing 86% normal corn silage (Control) or bm3 corn silage (BMCS). The remainder of the diets consisted of soybean meal, urea, monensin, vitamins, and minerals. Dry matter intake was greater (P < 0.01) for steers offered ad libitum access to BMCS than for those with ad libitum access to the Control diet. The BMCS treatment resulted in improved (P < 0.05) apparent total-tract digestibility of DM, OM, NDF, and ADF. Mean concentration of total VFA and molar proportions of acetate were increased (P < 0.05) by feeding BMCS. There tended to be a DMI x hybrid interaction (P = 0.16) for apparent total-tract digestibility of NDF. When diets were offered ad libitum, BMCS increased NDF digestibility by 10.5 percentage units compared with Control, but, when DMI was limited, BMCS increased NDF digestibility by 15.8 percentage units. In Trial 2, 128 steer contemporaries of those used in Trial 1 (245 +/- 13 kg) were offered ad libitum access to BMCS or Control diets as used in Trial 1. After a 112-d treatment period, concentrate in the diet was increased, and all steers were fed a common finishing diet. During the 112-d treatment period, steers receiving BMCS consumed 0.45 kg more DM/d (P < 0.05) and had similar ADG (P > 0.10), compared with those steers receiving the Control silage. This resulted in poorer (P < 0.01) feed efficiency for steers receiving BMCS. Finishing phase and overall performance of the steers was not different (P > 0.10) due to treatment. Although feeding BMCS in growth-phase diets resulted in increased daily DMI and improved digestibility of DM and fiber, it did not result in improved steer feedlot ADG compared with Control silage. PMID- 11063323 TI - Single daily intramuscular injections of low quantities of recombinant ovine interferon-tau extends luteal life-span in Angora goats. AB - The objective of this study was to determine whether single, daily intramuscular injections of low amounts of ovine interferon-tau (ovIFN-tau) would extend luteal life-span in nonpregnant Angora goats. Female goats were assigned randomly to receive a single daily injection of 1) PBS (control; n = 11), 2) 125 microg/d ovIFN-tau (n = 11), or 3) 500 microg/d ovIFN-tau (n = 11) from d 14 to 20 after estrus. Luteal life-span was defined as the number of days from the synchronized estrus until serum progesterone (P4) declined (< 0.5 ng/mL) and was of normal duration in controls (19.4 +/- 0.3 d) but was increased (P < 0.05) in goats receiving 125 microg/d (23.2 +/- 1.3 d) and 500 microg/d (25.5 +/- 1.2 d) ovIFN tau. Injection of either ovIFN-tau dose caused an initial decrease (P < 0.05) in serum P4 concentrations relative to controls but did not differ from controls thereafter. Rectal temperatures increased (P < 0.05) following ovIFN-tau treatment until d 18 for goats given the lower dose and throughout the treatment period for those given 500 microg/d. In summary, injections of as little as 125 microg/d of ovIFN-tau extended luteal life-span in goats. This dose caused a transient reduction in serum P4 concentrations and induced hyperthermia. PMID- 11063324 TI - Effects of supplemental zinc and manganese on ruminal fermentation, forage intake, and digestion by cattle fed prairie hay and urea. AB - One in vitro and one in vivo metabolism experiment were conducted to examine the effects of supplemental Zn on ruminal parameters, digestion, and DMI by heifers fed low-quality prairie hay supplemented with urea. In Exp. 1, prairie hay was incubated in vitro for 24 h with five different concentrations of supplemental Zn (0, 5, 10, 15, and 20 ppm) and two concentrations of supplemental Mn (0 and 100 ppm), both provided as chloride salts. Added Mn increased (P < 0.02) IVDMD, but added Zn linearly decreased (P < 0.03) IVDMD. Added Zn tended to increase the amount of residual urea linearly (P < 0.06) at 120 min and quadratically (P < 0.02) at 180 min of incubation, although added Mn counteracted these effects of added Zn. Six 363-kg heifers in two simultaneous 3 x 3 Latin squares were fed prairie hay and dosed once daily via ruminal cannulas with urea (45 or 90 g/d) and with Zn chloride to provide the equivalent of an additional 30 (the dietary requirement), 250, or 470 ppm of dietary Zn. After a 7-d adaptation period, ruminal contents were sampled 2, 4, 6, 12, 18, 21, and 24 h after the supplement was dosed. Supplemental Zn did not alter prairie hay DMI (mean = 4.9 kg/d) or digestibility, although 470 ppm added Zn tended to decrease (P < 0.06) intake of digestible DM, primarily due to a trend for reduced digestibility with 470 ppm supplemental Zn. Zinc x time interactions were detected for both pH (P = 0.06) and NH3 (P = 0.06). At 2 h after dosing, ruminal pH and ruminal ammonia were linearly decreased (P < 0.05; P < 0.01) by added Zn. At 5 h after feeding, ruminal pH was linearly increased (P < 0.05) by added Zn, suggesting that added Zn delayed ammonia release from urea. The molar proportion of propionate in ruminal fluid was linearly and quadratically increased (P < 0.02; P < 0.01) whereas the acetate:propionate ratio was linearly and quadratically decreased (P = 0.02; P < 0.05) by added Zn. Through retarding ammonia release from urea and increasing the proportion of propionate in ruminal VFA, Zn supplementation at a concentration of 250 ppm may decrease the likelihood of urea toxicity and increase energetic efficiency of ruminal fermentation. PMID- 11063325 TI - Intake, digestibility, and composition of orchardgrass and alfalfa silages treated with cellulase, inoculant, and formic acid fed to lambs. AB - The objectives of this study were to determine the effect of a cellulase (from Trichoderma longibrachiatum) alone or combined with a bacterial inoculant (Lactobacillus plantarum and Pediococcus cerevisiae) or formic acid on composition, intake, and digestibility of orchardgrass (Dactylis glomerata L.) and alfalfa (Medicago sativa L.) silages. Orchardgrass and alfalfa were harvested at the early heading stage and at the early bloom stage of maturity and wilted to approximately 22 and 32% DM, respectively. Forages were then ensiled in 100-L sealed barrels for at least 60 d before they were fed to lambs. Silage treated with cellulase had lower (P < .001) pH and lower (P < .001) acetic acid and NH3 N concentrations than untreated silage of both plant species and a higher (P = .004) lactic acid concentration than the control treatment of alfalfa silage. Fermentation characteristics of cellulase-treated silages, especially of alfalfa, were further enhanced by use of inoculant. Formic acid addition increased (P < .001), reducing sugar concentration of cellulase-treated orchardgrass and alfalfa silage by 90 and 154%, respectively, and decreased (P < .001) NH3 N concentration of cellulase-treated alfalfa silage by 19%. Averaged across plant species, cellulase, combined with inoculant or formic acid, resulted in 8 and 13% greater (P = .03) DMI, respectively, than the control silage. Extensive enzymatic cell wall degradation during ensiling decreased (P = .003) NDF intake of cellulase treated orchardgrass silage by 25% and decreased (P = .001) cellulose intake by 23%, when averaged across plant species. Addition of formic acid increased (P = .003) NDF intake of cellulase-treated orchardgrass silage by 19%. Averaged across species, cellulase application decreased (P < .05) silage NDF digestibility by 18%. Greater sugar and lower acetic acid, NH3 N, and NDF concentrations resulted in greater DMI of cellulase-treated silage than of control silage, when cellulase was combined with formic acid or inoculant. PMID- 11063326 TI - Rapid communication: nucleotide sequence of red seabream, Pagrus major, beta actin cDNA. PMID- 11063327 TI - Rapid communication: mapping of the Mannose-Binding Lectin 2 (MBL2) gene to pig chromosome 14. PMID- 11063328 TI - Rapid communication: nucleotide sequence of the ovine lipoprotein lipase cDNA. PMID- 11063329 TI - Lymphocyte homing to allografts. PMID- 11063330 TI - Liver preservation: a comparison of celsior to colloid-free University of Wisconsin solution. AB - INTRODUCTION: Celsior (CEL) was formulated specifically for heart preservation. Recently some preliminary reports have suggested that CEL is also effective for liver preservation. In this study liver preservation with CEL was compared to colloid-free University of Wisconsin solution (MUW). METHODS: Arterialized rat liver isografts were flushed and stored for 24 hr at 0 degrees C in CEL or MUW before orthotopic transplantation. Plasma albumin, bilirubin, glucose, aspartate aminotransferase, and alkaline phosphatase were measured 1, 2, 3, 7, 14, 21, and 28 days after surgery. RESULTS: All recipients of MUW-preserved livers survived, none of the recipients of CEL-preserved grafts lived beyond 3 days. On day 1, AST was raised in all rats but rats receiving CEL-preserved liver grafts were also markedly hypoglycemic, hypoalbuminemic and had elevated alkaline phosphatase. CONCLUSION: Celsior is not an effective solution for long-term liver preservation in its present composition. PMID- 11063331 TI - Effects of micro-encapsulation on morphology and endocrine function of cryopreserved neonatal porcine islet-like cell clusters. AB - BACKGROUND: For the success of clinical islets transplantation, the development of a long-term storage method is necessary. However, the structure of digested islets is scanty for culture and cryopreservation. In this study, the effect of micro-encapsulation to cryopreserved porcine islet-like cell clusters (ICCs) was investigated. METHODS: The ICCs prepared from neonatal pigs by collagenase digestion and culture technique were cryopreserved and micro-encapsulated in 5% agarose membranes. After cryopreservation, ICC cultured without encapsulation (group A) and cultured with encapsulation (group B) were assessed by comparison with no cryopreserved ICC (control) both in vitro by static incubation test and in vivo in a xenotransplantation study. RESULTS: Micro-encapsulation was able to maintain the fine morphology and the number of ICCs of group B after 7 days of culture. There were not significant differences in insulin secretion of group B and control on day 1 and 7 of culture (1 day:11+/-0.99, 7 days: 5.30+/-1.08 microU/ICC/hr NS versus control). On day 7 of culture, the retrieval rate of group B (105.2+/-9.8%) is obviously higher compared with group A (63.0+/-6.3%). In the xenotransplatation model, the ICCs of group B showed long survival time (7.9+/-0.4 weeks) and good transplantation effect. CONCLUSION: Our study suggests that micro-encapsulation is one of the useful method for cryopreserved ICC to maintain the fine morphology and effectively recover the endocrine function. PMID- 11063332 TI - The fate of liver grafts declined for subjective reasons and transplanted out of a local organ procurement organization. AB - BACKGROUND: Decisions made by transplant surgeons to decline liver grafts for local use are based on both objective and ill-defined subjective parameters. These livers may be offered and subsequently transplanted at non-local centers. We analyzed the fate of these exported livers, focusing on the outcome of grafts declined for subjective reasons. The aim is to determine whether local surgeons' concerns about inferior graft function are justified. METHODS: Over a 3-year period, 13.3% of 555 livers in our organ procurement organization (OPO) were exported and transplanted out of the local area. Donor data and reason for decline were obtained from an extensive OPO database. Objective reasons for decline were based on no appropriate matched recipient due to donor size, serologies, or malignancy with potential for spread. Subjective parameters were related to the procuring surgeon's assessment and included variables such as medical and social history, abnormal liver enzymes, older age, organ visualization, and biopsy. Recipient data were obtained from questionnaires sent to outside transplant centers. RESULTS: There was a significantly higher rate of nonfunction in the subjective group (17.1%), compared to the objective group (0%). One-year graft and patient survival were 79 and 83% for the objective group and 59 and 68% for the subjective group (P=NS). When donors declined for medical/social history were excluded from the subjective group, leaving only grafts declined based solely on the surgeon's assessment of graft quality, there is a significant difference in graft survival (79% for objective and 46% for this subjective subgroup, P=0.03). CONCLUSIONS: Livers declined for local use based on subjective assessment by the procuring surgeon have a high nonfunction rate, associated with a high morbidity. Therefore, the use of these grafts should be restricted to recipients at the most urgent status. PMID- 11063333 TI - Changes of urinary alpha1-microglobulin in the assessment of prognosis in renal transplant recipients. AB - BACKGROUND: After transplantation, even if the graft starts functioning immediately, there are morphological and functional changes in tubular structures. In addition, acute allograft rejection causes damage in the tubular epithelium, tubular basement membrane, and intertubular connective tissue. It also affects the functional capacity of proximal tubular cells resulting in impaired reabsorption and thus increased urinary excretion of low molecular weight proteins. METHODS: We present a double-antibody radioimmunoassay for determination of the concentration of alpha1-microglobulin (alpha1 M) in urine. It was used to measure urinary excretion of alpha1 M approximately once a week during the first 1-6 posttransplant weeks in 136 consecutive patients: 30 patients developing acute rejection (75 24-hr urine samples) and 106 patients with stable graft function (223 24-hr urine samples). The results are expressed as alpha1 M/creatinine ratios. RESULTS: Approximately 8 days after transplantation the mean (+/-SD) urinary alpha1 M/creatinine ratio of all patients was 17.0+/-14.8 mg/mmol, being about the same both in patients with uncomplicated posttransplantation course (16.3+/-14.0 mg/mmol) and in those who later developed rejection (19.3+/-15.1 mg/mmol), but about 60-fold higher than in healthy controls (0.27+/-0.15 mg/mmol). At that time, when all patients were included there was a correlation (r=0.3465, P<0.001) between alpha1 M/creatinine ratio and duration of cold ischemia. Thereafter, during the second week alpha1 M/creatinine ratio decreased in 89% of patients with stable graft function, but only in 14% of patients who later developed rejection (P<0.001). On the contrary, a significant increase (P<0.01) of alpha1 M/creatinine ratio was observed 4 to 1 day before rejection in all 15 patients, who had urines collected at that time. At the end of the follow-up period, alpha1 M/creatinine ratio in patients with rejection was 3-fold compared with the nonrejecting patients, and 100-fold compared with the healthy controls. CONCLUSION: These results show that cadaveric transplantation results in impaired low molecular weight protein reabsorption, the degree of dysfunction relating to the duration of cold ischemia, and suggest that during the posttransplant weeks decreasing alpha1 M/creatinine ratio in consecutively collected urine samples indicates improved tubular function and in most cases rules out development of acute rejection. PMID- 11063334 TI - Controlled non-heart-beating donor liver transplantation: a successful single center experience, with topic update. AB - BACKGROUND: The critical shortage of transplantable organs necessitates utilization of unconventional donors. We describe a successful experience of controlled non-heart-beating donor (NHBD) liver transplantation. METHODS: Controlled NHBDs had catastrophic head injury, prognosis for no meaningful recovery, decision to withdraw life support, and subsequent consent for donation. After stopping mechanical ventilation in the operating room, death determination by a nontransplant caregiver, and rapid aortic cannulation, liver and kidneys were recovered. RESULTS: Controlled NHBDs contributed 5% of hepatic allografts (8/164) from August 1996 through June 1999 (9% in 1998). Sixteen NHBDs afforded 8 livers and 24 kidneys. Liver donors (n=8) were 11-66 years old; half were >50 years old. Premortem alanine aminotransferase was 25-157 U/L. Arrest occurred 3 27 min after stopping ventilation. Perfusion started 3-5 min after incision, and <22 min after hypotension (mean arterial pressure: <50 mmHg). Patient and graft survivals are 100% at 18+/-12 months follow-up. There was no intraoperative complication, reperfusion syndrome, poor graft function, primary nonfunction, arterial thrombosis, biliary complication, or serious infection. Postoperative day 2 prothrombin time was 13+/-1 sec. Peak alanine aminotransferase was 980+/ 601 U/L. Intensive care unit and posttransplant lengths of stay were 2+/-2 and 10+/-7 days, respectively. Soon after transplantation there was frequent temporary hyperbilirubinemia (five of eight recipients; bilirubin peak: 7-29 mg/dl, 2-3 weeks after transplantation) and rejection (4/8 recipients, <3 weeks after transplantation). CONCLUSIONS: NHBDs significantly and safely expanded our donor pool. NHBD surgeons must be capable of rapid procurement. Cautious liberalization of criteria for accepting livers from NHBDs with confounding risk factors is justified. Refined ethics guidelines would broaden approval of NHBDs. PMID- 11063335 TI - A prospective study of the natural course of cytomegalovirus infection and disease in renal allograft recipients. AB - BACKGROUND: Cytomegalovirus (CMV) infection is the single most frequent infectious complication in renal transplant recipients. Because no CMV prophylaxis is given and ganciclovir is used only as deferred therapy for CMV disease at our center, we have been able to study the natural course of CMV infections. The aim was to assess risk factors for CMV infection and disease and thus identify subgroups of patients likely to benefit from CMV prophylaxis or preemptive therapy. METHODS: Between October 1994 and July 1997, 477 consecutive renal transplant recipients (397 first transplants and 80 retransplants) were included in the study. The patients were followed prospectively for 3 months with serial measurements of CMV pp65 antigen for monitoring activity of CMV infections. RESULTS: The incidence of CMV infections in first transplants was 68% in D+R- and D+/-R+ serostatus groups, whereas the incidence of CMV disease was higher in D+R- (56%) than in D+/-R+ (20%, P<0.001). No difference in severity of CMV disease in D+R- and D+/-R+ was seen except for an increased incidence of hepatitis in primary infections. One of 14 deaths could be associated with CMV disease in a seropositive recipient. Cox regression analysis showed that rejection (RR 2.5, P<0.01) and serostatus group D+R- (RR 3.9, P<0.001) were significant risk factors for development of CMV disease. The maximum CMV pp65 antigen count had significant correlation to disease only in CMV seropositive recipients, P<0.001. Conclusion. Renal transplant recipients can safely be given deferred ganciclovir therapy for CMV disease if they are intensively monitored for CMV infection. Patients with primary CMV infection (D+R-), CMV infected patients undergoing anti-rejection therapy and R+ patients with high CMV pp65 counts seem to have a particular potential for benefit from preemptive anti-CMV therapy. PMID- 11063336 TI - Long-term oral ganciclovir prophylaxis for prevention of cytomegalovirus infection and disease in cytomegalovirus high-risk renal transplant recipients. AB - BACKGROUND: Although specific therapy is available with ganciclovir, cytomegalovirus (CMV) disease remains a major problem after renal transplantation especially in CMV seronegative recipients of organs of seropositive donors (D+R ). METHODS: In an open-labeled prospective controlled trial we evaluated the effect of long-term oral ganciclovir prophylaxis (3 g/day for 3 months posttransplantation) in a cohort of 31 CMV-high risk (D+R-) renal transplant recipients (GC) compared with a cohort of 28 high-risk patients with targeted CMV prophylaxis (CO) receiving i.v. ganciclovir during antirejection therapy. Primary end-points were CMV infection, diagnosed by pp65 antigenemia assay or serologic method, and CMV disease. Additionally severity of CMV disease quantified by a scoring system was evaluated. RESULTS: CMV prophylaxis significantly reduced the incidence of CMV infection (CO: 75%, GC: 45%; P<.05) and CMV disease (CO: 60%, GC: 29%; P<.05) without relevant side effects and without any clinical suspicion of ganciclovir resistance. Severity of CMV disease as quantified by a scoring system was reduced from 8.3+/-6.7 points in controls to 3.3+/-2.6 points in ganciclovir-treated patients (P<.05). Mortality did not differ significantly between the two groups (CO: n=3, GC: n=1; NS). However, there was one lethal CMV disease and a second death possibly attributable to CMV disease in the control group, whereas in ganciclovir-treated patients there was no CMV-associated fatal outcome. CONCLUSION: Long-term oral ganciclovir prophylaxis is effective and safe in CMV high-risk renal transplant recipients. PMID- 11063337 TI - Reproductive hormones after pancreas transplantation. AB - BACKGROUND: Reproductive hormone function after pancreas transplantation (PTX) is unknown as it has not been studied. METHODS: We prospectively studied PTX recipients to determine changes in reproductive hormones after PTX. Testosterone or estradiol, leutinizing hormone, follicle stimulating hormone, and prolactin were determined before and 1 year after PTX in 23 patients (10 women, 13 men) followed for more than 1 year after PTX. Of these, 11 received simultaneous kidney-PTX; 8 PTX only; and 4, PTX after kidney. Average age was 38.4+/-1.6 years and average duration of diabetes was 24.5+/-1.3 years. Nine (four women, five men) patients had been on dialysis pre-PTX. Sixteen of 23 patients were treated with cyclosporine and seven with FK-506 along with prednisone and azathioprine post-PTX. RESULTS: Mean testosterone in men was normal pre- and post-PTX. Two men had secondary hypogonadism pre-PTX with resolution in one and persistence in the other post-PTX. Five of the ten women had evidence of hypogonadism pre-PTX: three had primary hypogonadism and two had secondary hypogonadism. Post-PTX, 7 of 10 women had abnormal reproductive hormones: 4 had primary hypogonadism, 2 had secondary hypogonadism, and 1 developed hyperestrogenemia with elevated estradiol (482 pg/ml) and leutinizing hormone (41 IU/liter). Mean prednisone dose and cyclosporine trough level were higher in the women than the men (P<0.05). No cases of secondary hypogonadism that developed or resolved post-PTX were related to changes in prolactin, renal function, or hyperglycemia. CONCLUSIONS: Women are more likely than men to have reproductive hormone abnormalities pre- and post-PTX and the causes may be multiple. PMID- 11063338 TI - Abstension from treatment of low-level pp65 cytomegalovirus antigenemia after liver transplantation: a prospective study. AB - BACKGROUND: Ganciclovir is a highly effective and relatively safe drug to treat cytomegalovirus (CMV) infection in liver transplant patients; CMV resistance to ganciclovir is progressively emerging due to the extensive use of the drug in transplant and AIDS patients; CMV pp65 antigenemia allows early diagnosis of CMV infection and quantitation of the viral load; preemptive antigenemia-guided therapy of CMV infection can prevent CMV disease but the threshold of antigenemia value above which treatment has to be instituted is unclear. METHODS: To demonstrate the safety of abstention from preemptive treatment in the presence of low levels of antigenemia 77 consecutive liver transplant recipients were prospectively evaluated. Antigenemia was tested twice a week from transplantation until discharge, then once a week until the third postoperative month. In absence of risk factors for CMV disease, namely donor positive/recipient negative CMV serology, treatment with antibodies to lymphocytes and retransplantation, only patients with antigenemia of more than 50 or symptoms possibly related to CMV infection had preemptive treatment. RESULTS: A total of 32 patients had at least one positive antigenemia test with a value less than 50; 22 (68.7%) spontaneously cleared the virus, 3 were treated with i.v. ganciclovir for the presence of fever, and the other 7 (21,8%) progressed to values of antigenemia of more than 50 and were treated even if asymptomatic. No CMV disease was observed in these patients. CONCLUSION: CMV antigenemia less than 50 in liver transplant recipients with low and intermediate risk for CMV disease does not mandate preemptive ganciclovir treatment. Close surveillance with repeated determination of antigenemia until its negativization and careful clinical and laboratory monitoring is advisable. PMID- 11063339 TI - Expression of angiotensin II type I receptor on erythroid progenitors of patients with post transplant erythrocytosis. AB - BACKGROUND: The pathogenesis of posttransplant erythrocytosis (PTE) has been elusive. Angiotensin converting enzyme inhibitors (ACEI) are efficacious in lowering the hematocrit of patients with PTE and angiotensin II (AII) type I receptors (AT1R) were recently detected on red blood cell precursors, burst forming unit-erythroid- (BFU-E) derived cells. The purpose of this study was to determine whether there is increased expression of the AT1R on BFU-E-derived cells of patients with PTE, which might contribute to the pathogenesis of PTE. METHODS: Twelve healthy volunteers and 25 transplant recipients (13 patients with and 12 without PTE) were studied. BFU-E from peripheral blood were cultured in methylcellulose and BFU-E-derived colonies were harvested on day 10. Western blotting was used to detect AT1R and erythropoietin receptor (EpoR) expression. Intracellular free calcium in response to AII and erythropoietin (Epo) was measured with digital video imaging. RESULTS: There were no differences between transplant patients, with and without PTE, with respect to weight, age, sex, blood pressure, serum creatinine, circulating renin, angiotensin II, and Epo levels. Hematocrit, red blood cell number, BFU-E-derived colony number,and size were significantly increased in PTE compared with other two groups. AT1R expression was increased by 44% on the erythroid progenitors of PTE versus non posttransplant erythrocytosis patients and by 32% in PTE patients versus normal volunteers. AT1R expression correlated significantly with the hematocrit in PTE (Spearman r=0.68, P=0.01). In contrast, EpoR expression was equivalent in all groups. The AT1R was functional since a significant increase in [Ca(i)] was observed in Fura-2 loaded day 10 cells when stimulated with AII (182%, P<0.0001). CONCLUSION: An increase in AT1R density was observed in erythroid precursors of transplant patients with PTE compared to those without PTE and normal volunteers, and the level of AT1R expression in PTE correlated significantly with the hematocrit. In contrast, EpoR expression was not different in PTE compared with non posttransplant erythrocytosis or normal controls. This study supports a role for the AT1 receptor signaling pathway in the pathogenesis of PTE. PMID- 11063340 TI - The prevalence of TT virus infection in renal transplant recipients in Brazil. AB - BACKGROUND: Recently, TT virus (TTV) was discovered as a potential causative agent for non-A-E hepatitis. Little is known about the prevalence of TTV infection in renal transplant recipients. METHODS: One hundred and seventeen Brazilian renal transplant recipients and 100 normal subjects were examined to determine the prevalence of TTV infection. The TTV DNA in serum and its genotype were examined using polymerase chain reaction and restriction enzyme length polymorphism, respectively. RESULTS: TTV DNA was detected in 63/117 (53.8%) renal transplant recipients in contrast to its detection in 10/100 (10%) normal subjects (P<0.001). There was no statistical difference in the distribution of TTV genotypes between these groups. There was no significant difference in clinical backgrounds between TTV positive and negative patients. CONCLUSIONS: These results indicate a risk for TTV infection in renal transplant recipients in Brazil. They also indicate that TTV itself might not have a strong correlation with the pathogenicity of liver diseases. PMID- 11063341 TI - Role of platelet-activating factor in functional alterations induced by xenoreactive antibodies in porcine endothelial cells. AB - BACKGROUND: Platelet-activating factor (PAF) is a phospholipid mediator of inflammation which has been implicated in rejection. The interaction of anti alpha-galactosyl natural antibodies (anti-alpha gal Abs) with endothelial cells is the initial step for the development of xenograft rejection. In our study, we stimulated porcine aortic endothelial cells (PAEC) with anti-alpha gal IgG to investigate the synthesis of PAF from PAEC and its biological consequences. METHODS AND RESULTS: PAF was extracted and chromatographically purified from cultured PAEC stimulated with baboon anti-alpha gal Abs. The Abs induced a dose dependent synthesis of PAF peaking after 30 min of incubation, and decreasing thereafter. Concomitant cell shape change, motility, and cytoskeleton redistribution were observed. These events were prevented by addition of a panel of PAF-receptor antagonists. An SV40 T-large antigen-immortalized PAEC line was engineered to express PAF acetyl-hydrolase (PAF-AH) cDNA, the major PAF inactivating enzyme. These transfected cells exposed to anti-alpha gal Abs showed reduced cell contraction and motility compared with empty vector-transfected cells. Moreover, in PAEC stimulated with anti-alpha gal Abs, the synthesis of PAF promoted the adhesion of a monocytic cell line as shown by the inhibitory effect of PAF-receptor antagonists and of PAF-AH expression. Finally, studies on cell monolayer demonstrated an enhanced permeability 48 hr after exposure to anti alpha gal Abs, and this increase was prevented by PAF-inactivation and by PAF receptor blockade. CONCLUSIONS: These results demonstrate that on stimulation with anti-alpha gal Abs, PAEC synthetize PAF which can contribute to several vascular events involved in xenograft rejection. PMID- 11063342 TI - Fucosyl transferase (H) transgenic heart transplants to Gal-/- mice. AB - BACKGROUND: We have previously described the rejection of Gal+ mouse hearts by mice lacking Gala(1,3)Gal (Gal-/-) and demonstrated this to be a model of xenogeneic hyperacute rejection (HAR) which would occur in pig-to-human/primate xenotransplantation, where Gal+ antibody (Ab) and complement (C') mediate HAR. To reduce the amount of Gal present we used fucosyl transferase (H) as a transgene, H transferase competes for the same substrate as Gal transferase and reduces Gal expression by >90%. METHODS: Gal-/- mice received a heart graft from C57BL/6 Gal+ or H transgenic mice and additional Gal Ab and C' provided; HAR was monitored by direct observation for up to 90 min, or by palpation thereafter. When grafts were rejected they were examined macro- and microscopically. RESULTS: H transgenic mice were used as donors to Gal-/- mice; it was found that: 1) C57BL/6 or H transgenic hearts were not rejected by Gal-/- recipients within 90 min in the absence of additional Gal Ab. 2) If additional Gal Ab and C' were provided as fresh normal human serum (NHS), Gal+ (C57BL/6) grafts were rejected by Gal-/- mice in approximately 34 min, whereas H transgenic hearts mostly lasted up to 17 hr, but were then rejected. The histological appearances showed features of both Arthus and Shwartzmann phenomena. 3) Mice hyperimmunized with Gal with anti-Gal titers of >1:20,000, rejected Gal+ grafts in 31 min; the survival was prolonged to 75 min with the H transgenic hearts. CONCLUSION: The presence of the H transgene in donor hearts transplanted to naive Gal-/- mice delays the onset of HAR, but rejection ultimately occurs; if the mice are hyperimmune earlier rejection occurs. The expression of the H transgene alone is insufficient to avoid HAR in the Gal-/- mouse model; the presence of other transgenes and techniques will be required to give an appropriate increase in survival of pig-to human/primate grafts. PMID- 11063343 TI - Evaluation of human cytomegalovirus gene expression in thoracic organ transplant recipients using nucleic acid sequence-based amplification. AB - BACKGROUND: Human cytomegalovirus (CMV) infection is a major cause of morbidity in transplant patients. Early diagnosis and treatment have been shown to improve outcome. We evaluated the suitability of CMV immediate early, early, and late gene expression detected by nucleic acid sequence-based amplification (NASBA) as markers of CMV infection. METHODS: Blood samples were taken immediately before transplant and every one to two weeks after transplantation for 12 weeks from 50 patients undergoing thoracic organ transplantation. CMV-NASBA was performed and results compared with serology, CMV pp65 antigenaemia (CMV-AG) and the development of clinical CMV infection. Patients received "preemptive" anti-CMV therapy with ganciclovir based on the CMV-AG results. RESULTS: CMV immediate early and early gene expression were detected in 87 and 47%, respectively, of patients without other evidence of CMV infection. CMV late gene expression had a sensitivity of 97% for infection (compared with 83% for CMV-AG P=0.06) and a specificity of 93% (compared with 100% P=NS). Late gene expression occurred at the same time as CMV antigenaemia but 1.1 weeks earlier than the threshold of antigenaemia (CMV-AG>10) used to initiate preemptive therapy. CONCLUSION: NASBA provided a standardized tool for the detection of CMV transcripts with a greater sensitivity than the standard antigenemia test. Detection of immediate early and early gene transcripts was not specific for subsequent infection. CMV late gene expression determined by NASBA was an accurate and early marker of CMV infection. Detection of CMV late gene expression could be used to trigger "preemptive" anti CMV therapy. PMID- 11063344 TI - Suppression of primary T-cell responses and induction of alloantigen-specific hyporesponsiveness in vitro by the Janus kinase inhibitor tyrphostin AG490. AB - BACKGROUND: Tyrphostin AG490 has recently been shown to block interleukin (IL)-2 receptor gamma-chain-associated Janus kinase 3. Here, we analyzed the effect of AG490 on T-cell alloresponses in vitro. METHODS: For the evaluation of T-cell activation, DNA synthesis, surface marker expression, cytokine secretion, intracellular calcium mobilization, early protein tyrosine phosphorylation, and apoptosis were measured. RESULTS: AG490 effectively inhibited T-cell proliferation in human mixed lymphocyte culture (MLC) even when added 4 days after culture initiation. Inhibition of IL-2-dependent proliferation in T-cell blasts and the incapability of IL-2 or IL-15 to restore proliferation in AG490 treated MLC suggests interference with cytokine receptor signaling. T-cell receptor-triggered early protein tyrosine phosphorylation, calcium mobilization, up-regulation of CD69, and initial CD25 expression were not affected. Interestingly, AG490 substantially inhibited production of IL-2 and interferon gamma in T cells stimulated with alloantigen or via CD3 and CD28. In CD28 independent activation models (e.g., stimulation with phorbol myristate acetate plus ionomycin), however, cytokine secretion was not inhibited. Pretreatment of primary MLC with AG490 resulted in substantial down-regulation of secondary responses to cells from the original donor as opposed to third-party cells or phytohemagglutinin. Unresponsiveness was induced also in T cells stimulated with CD3 monoclonal antibody. Induction of apoptosis in polyclonally activated T cells and the incapability of IL-2 to reverse specific hyporesponsiveness, suggest programmed cell death as an important mechanism underlying antigen-specific down regulation of alloresponses. CONCLUSIONS: We demonstrate that AG490 blocks different manifestations of T-cell activation. This and its ability to induce alloantigen-specific hyporesponsiveness point to a potential use for interfering with alloreactivities in vivo. PMID- 11063345 TI - Cytokine and chemokine expression kinetics after corneal transplantation. AB - BACKGROUND: Allogeneic rejection is the most common cause of corneal graft failure. The aim of this work was to establish the kinetics of cytokine and chemokine mRNA expression before and after onset of corneal graft rejection. METHODS: Intracorneal cytokine and chemokine mRNA levels were investigated in the Brown Norway-->Lewis inbred rat model in which rejection onset is observed at 8/9 days after grafting in all animals. Nongrafted corneas and syngeneic (Lewis- >Lewis) corneal transplants were used as controls. Donor and recipient cornea was examined by quantitative competitive reverse transcription-polymerase chain reaction (RT-PCR) for hypoxyanthine phosphoribosyltransferase (HPRT), CD3, CD25, interleukin (IL)-1beta, IL-1RA, IL2, IL-6, IL-10, interferon-gamma (IFN-gamma), tumor necrosis factor (TNF), transforming growth factor (TGF)-beta1, and macrophage inflammatory protein (MIP)-II and by nonquantitative RT-PCR for IL4, IL-5, IL-12 p40, IL-13, TGF-beta2, monocyte chemotactic protein-1 (MCP-1), MIP 1alpha, MIP-1beta, and RANTES (for regulated upon activation normal T cell expressed and secreted). RESULTS: A biphasic expression of cytokine and chemokine mRNA was found after transplantation. During the early phase (days 3-9), there was an elevation of the majority of the cytokines examined, including IL-1beta, IL-6, IL-10, IL-12 p40, and MIP-II. There was no difference in cytokine expression patterns between allogeneic or syngeneic recipients at this time. In syngeneic recipients, cytokine levels reduced to pretransplant levels by day 13, whereas levels of all cytokines rose after observed rejection onset in the allografts, including TGF-beta1, TGF-beta2, and IL-1RA. The T cell-derived cytokines IL-4, IL-13, and IFN-gamma were detected only during the rejection phase in allogeneic recipients. CONCLUSIONS: There is an early cytokine and chemokine response to the transplantation process, evident in syngeneic and allogeneic grafts, that probably drives angiogenesis, leukocyte recruitment, and affects leukocyte functions. After an immune response has been generated, allogeneic rejection results in the expression of Th1 cytokines (IL-2, IL-12 p40, IFN-gamma), Th2 cytokines (IL-4, IL-6, IL-10, and IL-13), and antiinflammatory/Th3 cytokines (TGF-beta1/2 and IL-1RA). PMID- 11063346 TI - Acute tubular necrosis induced by high level of cyclosporine A in a lung transplant. AB - We report a case of acute accidental cyclosporine A intoxication in a lung transplant patient. The intoxication led to renal failure due to acute tubular necrosis, which was partially reversible. A review of the literature on the renal consequences of cyclosporine A intoxication is given. PMID- 11063347 TI - Domino liver transplantation from a living related donor. AB - BACKGROUND: Although domino liver transplantations (OLT) from cadaveric donors have been performed in about 50 cases since 1995, only one case in the Japanese literature has been reported on a domino OLT from a living related donor. The difficulties of the later surgery lie in the small size of the graft volume and the short length of the vascular cuffs in the graft. METHODS: The left lobe graft was procured from a 43-year-old younger brother of a familial amyloidotic polyneuropathy (FAP) patient. Next, the left lobe graft (510 g, 44% of the estimated standard liver volume of the FAP patient) was implanted into the 48 year-old female FAP patient. At surgery for the FAP patient, a sufficient length of the vascular cuffs was secured by an extended left lobe resection, although the right lobe graft was able to maintain sufficient vascular cuffs. The right lobe graft (720 g, 54% of the recipient's estimated standard liver volume) was then implanted in the 43-year-old male patient with liver cirrhosis and hepatocellular carcinoma (stage IV-A). RESULTS: The two recipients were discharged from the hospital 1 month after OLT. At 7 months after OLT, they are both doing well and the domino recipient is free of any tumor recurrence. CONCLUSION: A domino OLT from the living related donor can therefore be done safely when careful attention is paid to the graft volume and the length of the vascular cuffs for anastomosis. PMID- 11063348 TI - Renal transplant patient compliance with free immunosuppressive medications. AB - BACKGROUND: Noncompliance with immunosuppressive medications after renal transplantation is believed to be a major cause of allograft rejection and graft loss, with the impressive costs of these agents considered a significant reason for noncompliance. Our purpose was to determine the compliance rates of renal transplant patients who received their immunosuppressant therapy free of charge and evaluate their patterns of compliance. METHODS: All patients who received a renal transplant and received their immunosuppressant medications at our institution for their first year posttransplant were included in the study. Compliance rate was calculated and serum immunosuppressant concentrations were obtained to validate compliance assessments. RESULTS: Eighteen patients were included in the study. Approximately 48% of noncompliant patients were found to have subtarget drug concentrations, although only 14% of compliant patients had subtarget levels (chi2=12.9, P<0.001). At 5 months posttransplant, 95% of the patients remained compliant; however, by 12 months posttransplant, only 48% of the patients remained compliant. The mean time to the first noncompliant month was 9.8 months (95% confidence intervals=8.60-11.0). CONCLUSIONS: Patients who received their immunosuppressants free of charge were generally compliant within their first year of transplantation, however, compliance tended to decrease over time. This suggests that drug cost alone does not explain noncompliant behavior. Intensive efforts to increase medication compliance before month 8 posttransplantation should be implemented. PMID- 11063349 TI - Conversion to rapamycin immunosuppression in renal transplant recipients: report of an initial experience. AB - BACKGROUND: The aim of this study is to evaluate the effects of RAPA conversion in patients undergoing cyclosporine (CsA) or tacrolimus (Tac) toxicity. METHODS: Twenty renal transplant recipients were switched to fixed dose rapamycin (RAPA) (5 mg/day) 0 to 204 months posttransplant. Drug monitoring was not initially used to adjust doses. The indications for switch were chronic CsA or Tac nephrotoxicity (12), acute CsA or Tac toxicity (3), severe facial dysmorphism (2), posttransplant lymphoproliferative disorder (PTLD) in remission (2), and hepatotoxicity in 1. Follow-up is 7 to 24 months. RESULTS: In the 12 patients switched because of chronic nephrotoxicity there was a significant decrease in serum creatinine [233+/-34 to 210+/-56 micromol/liter (P<0.05) at 6 months]. Facial dysmorphism improved in two patients. No relapse of PTLD was observed. Five patients developed pneumonia (two Pneumocystis carinii pneumonia, one infectious mononucleosis with polyclonal PTLD lung infiltrate) and two had bronchiolitis obliterans. There were no deaths. RAPA was discontinued in four patients, because of pneumonia in two, PTLD in one, and oral aphtous ulcers in one. RAPA levels were high (>15 ng/ml) in 7 of 13 (54%) patients. CONCLUSIONS: RAPA conversion provides adequate immunosuppression to enable CsA withdrawal. However, when converting patients to RAPA drug levels should be monitored to avoid over-immunosuppression and adequate antiviral and Pneumocystis carinii pneumonia prophylaxis should be given. PMID- 11063350 TI - Optimal timing for a pancreas transplant after a successful kidney transplant. AB - BACKGROUND: For certain uremic, diabetic patients, a sequential transplant of a kidney (usually from a living donor) followed by a cadaver pancreas has become an attractive option. But how long to wait after the kidney transplant before proceeding with a pancreas transplant is unclear. We studied outcomes in recipients of a pancreas at varying times after a kidney to determine the optimal timing for the second transplant. METHODS: We compared pancreas after kidney (PAK) transplants performed early (< or =4 months) and late (>4 months) after the kidney transplant to determine any significant differences in surgical complications or outcomes between the two groups. RESULTS: Between January 1, 1994, and September 30, 1998, we performed 123 cadaver PAK transplants. Of these, 25 (20%) were early and 98 (80%) were late. Characteristics of the two recipient groups were similar. We found no significant differences in outcome between the two groups. The incidence of surgical complications (bleeding, leaks, thrombosis, infections) and of opportunistic infections (such as cytomegalovirus) did not significantly differ between the two groups. Graft and patient survival rates were also equivalent (P=NS). The incidence of acute rejection by 3 months posttransplant was 20% in both groups. CONCLUSION: The timing of the pancreas transplant for PAK recipients does not seem to influence outcome. As long as an acceptable organ is available and the recipient is clinically stable, a PAK transplant can be performed relatively soon after the kidney transplant. PMID- 11063351 TI - Mycophenolate mofetil, an alternative to cyclosporine A for long-term immunosuppression in kidney transplantation? AB - BACKGROUND: Mycophenolate-mofetil (MMF) is a nonnephrotoxic immunosuppressant most often used in combination with cyclosporine A (CsA) and prednisone (Pred). This study reports the outcome of 17 adult renal recipients whose immunosuppressive regimen was changed from CsA-Pred to MMF-Pred because of CsA nephrotoxicity. METHODS: CsA nephrotoxicity was diagnosed in all patients based on suggestive histopathological lesions on a renal biopsy. Sixteen patients had deteriorating graft function and 1 had isolated persistent proteinuria. Immunosuppressive therapy was changed 57+/-32 months posttransplant. RESULTS: After replacement of CsA by MMF, a reduction in serum creatinine was observed in all patients (mean 26+/-17%). This reduction was maintained 20+/-8 months after the change in therapy without any episodes of acute rejection. Serum lipids and blood pressure also decreased significantly. CONCLUSION: This study demonstrates that MMF-Pred can be an effective long-term immunosuppressive treatment alternative for renal transplant patients experiencing CsA nephrotoxicity. Such treatment may result in improved graft function, and better control of hypertension and hyperlipidemia. PMID- 11063352 TI - Exsanguinous metabolic support perfusion--a new strategy to improve graft function after kidney transplantation. AB - BACKGROUND: The compounding damage of warm ischemia (WI) followed by cold preservation is a major barrier in renal transplantation. Although the relative effect of WI is not yet well understood, therapeutic strategies have mostly focused on minimizing the pathology seen upon reperfusion from the cold. Our study was designed to examine the effect of restoration of renal metabolism by warm perfusion on graft survival and to investigate the compounding damage of WI. METHODS: Using a known critical canine autotransplantation model (1), kidneys were exposed to 30 min WI followed by 24 hr cold storage in Viaspan. They were then either reimplanted directly or first transitioned to 3 hr of warm perfusion with an acellular perfusate before reimplantation. Contralateral kidneys were subjected to 0, 30, or 60 min WI; 24 hr cold storage, and 3 hr warm perfusion. RESULTS: Transplanted kidneys that were warm perfused before reimplantation had both lower 24 hr posttransplant serum creatinine (median of 3.2 vs. 4.1 mg/dl) and lower peak serum creatinine (median of 4.95 vs. 7.1 mg/dl). Survival rate for warm perfused kidneys was 90% (9/10) vs. 73% (8/11). In the contralateral kidneys, metabolism was affected by the compounding damage of WI. Renal oxygen and glucose consumption diminished significantly, whereas vascular resistance and lactate dehydrogenase-release rose significantly with increasing WI. CONCLUSIONS: The results demonstrate a reduction of reperfusion damage by an acellular ex vivo restoration of renal metabolism. Furthermore, data from the contralateral kidneys substantiates the relative role of WI on metabolism in renal transplantation. PMID- 11063353 TI - Massive cerebral edema after I.V. cyclosporin overdose. AB - We describe a patient who accidentally received an infusion of cyclosporin at a rate of 30 mg/hr during 13 hr instead of the prescribed dose of 3 mg/hr and who concomitantly developed massive intracerebral edema with brainstem compression and death. A cyclosporin level as high as 1700 ng/ml could have been reached before the drug was withdrawn. To the best of our knowledge, this is the first case of fatal cyclosporin overdose reported in an adult patient. PMID- 11063354 TI - The relevance of donor T cell-directed immunoglobulin G in historic sera in the age of flow cytometry. AB - BACKGROUND: Renal transplant recipients with a positive historic cross-match due to donor T cell-directed IgG antibodies are considered to have decreased graft survival, even if their current serum is negative prior to transplantation. With the use of flow cytometric cross-match for testing current sera, false-negative results could be eliminated and the outcome of transplantation in this group of patients could be improved, assuming that immunological memory is effectively controlled with immunosuppression. METHODS: We reviewed our records to identify those patients who underwent cadaveric renal transplant, with a historic IgG positive cytotoxic T cell cross-match and a current negative flow cytometric T cell cross-match. RESULTS: Eighteen patients underwent cadaveric renal transplant in the face of a historic IgG positive T cell cross-match and a current negative flow cytometric T cell cross-match. In 14 patients treated with cyclosporine based immunosuppression the 1-, 2-, and 3-year cumulative graft survival rates were 57, 50, and 43%, respectively. Ten of the 14 patients (71%) ultimately lost their grafts. CONCLUSIONS: Even with negative flow cytometric cross-match in current serum, a positive historic conventional cross-match suggests a high risk of graft failure. PMID- 11063355 TI - Clotrimazole inhibits lung fibroblast proliferation in vitro: implications for use in the prevention and treatment of obliterative bronchiolitis after lung transplantation. AB - BACKGROUND: Immunosuppressive therapy has limited activity against the mesenchymal cell proliferation of obliterative bronchiolitis. Clotrimazole (CLT) has been shown to inhibit proliferation in normal and cancer cell lines. Here we investigate whether CLT inhibits the proliferation of lung mesenchymal cells. METHODS: Proliferation of human lung fibroblasts (MRC-5) in the presence of CLT was determined by [3H]thymidine incorporation. Messenger ribonucleic acid (mRNA) expression of platelet-derived growth factor (PDGF)-B and transforming growth factor (TGF)-beta after treatment with CLT was measured by reverse transcriptase polymerase chain reaction. RESULTS: Treatment of MRC-5 cells with CLT resulted in a significant reduction in proliferation as assessed by DNA incorporation and cell counts compared with dimethylsulfoxide alone. There was no cytotoxic effect associated with CLT treatment. Reverse transcriptase-polymerase chain reaction demonstrated a marked decrease in PDGF-B and TGF-beta mRNA levels in cells treated with CLT compared with those treated with dimethylsulfoxide. CONCLUSION: CLT inhibits proliferation of human lung fibroblasts. This inhibitory effect is associated with decreased levels of PDGF-B and TGF-beta mRNA expression and may have value in the prevention and treatment of obliterative bronchiolitis. PMID- 11063356 TI - Utility of laboratory testing for infants with seizures. AB - OBJECTIVES: Study objectives were to 1) determine the frequency with which laboratory studies are obtained, 2) determine the proportion of results that are clinically significantly abnormal, and 3) define the clinical characteristics of those with abnormal results, among infants with nonfebrile seizures (NFSz). METHODS: A retrospective consecutive cohort study of infants < or = 12 months old presenting to the ED of a tertiary care, children's hospital following a seizure. A 2-year review was performed. Serum chemistry results were classified as "normal," "outside of the normal range," or "clinically significantly abnormal." RESULTS: Sixty-seven of 134 (50%) with a NFSz were tested compared to 19/80 (24%) with a febrile seizure (FSz, P < 0.001). Nine (5 with hyponatremia and 4 with hypocalcemia) of the 67 (13%) tested NFSz infants had a clinically significant abnormality, as did 9 of 21 (43%) NFSz infants who seized in the ED compared to 0/46 (0%) without ED seizure activity (P < 0.0001). Hypothermia (T < 36.5 degrees C) and age less than 1 month were common characteristics of infants with clinically significant abnormalities. CONCLUSIONS: This is one of the only studies to have assessed the utility of laboratory testing for infants with seizures. Abnormal serum chemistries accounted for a greater proportion of seizures among this cohort compared to that reported previously for older children. Laboratory testing is recommended for NFSz infants who 1) are actively seizing in the ED, 2) have a temperature below 36.5 degrees C, or 3) are less than 1 month of age. PMID- 11063357 TI - Potential impact of a computerized system to report late-arriving laboratory results in the emergency department. AB - BACKGROUND: Results of some laboratory tests for Emergency Department (ED) patients return hours to days after the patient is discharged. Inadequate follow up for these late-arriving results poses medical and legal risks. We have developed, but not yet implemented, a computerized system called the Automated Late-Arriving Results Monitoring System (ALARMS). ALARMS scans the hospital's laboratory and ED registration databases to generate an electronic daily log of all late-arriving abnormal laboratory results for ED patients. OBJECTIVE: To determine the potential impact of ALARMS by assessing our ED's current quality of documented follow-up of late-arriving laboratory results. METHODS: We applied ALARMS retrospectively, to find all abnormal late-arriving laboratory results returned between 5/1/96 and 4/30/98 for ED patients for the following three tests: serum lead levels, Chlamydia cultures, or urine pregnancy tests. Medical records were reviewed for documentation of follow-up, which was considered appropriate if a clinician noted the abnormal result and documented a follow-up plan within 1 week after the result became available. Medical records were also reviewed for any evidence of complications attributable to delayed or inadequate follow-up. RESULTS: Over the 2-year study period, no appropriate follow-up was documented in 6/18 (33%) cases of elevated lead levels, 3/4 (75%) cases of late arriving positive pregnancy tests, and 23/39 (59%) cases of positive Chlamydia cultures. One case of a positive Chlamydia culture, for which there was no documented follow-up, was associated with subsequent development of pelvic inflammatory disease. CONCLUSION: Our current system of documented follow-up for late-arriving laboratory results has deficiencies. ALARMS, a computerized system of alerts for emergency physicians, has the potential to substantially improve documented follow-up of late-arriving laboratory results in the ED. PMID- 11063358 TI - Alcohol and injury in adolescents. AB - OBJECTIVE: To determine the frequency of alcohol ingestion in adolescent victims of major trauma and determine whether alcohol ingestion is associated with increased injury severity or death. METHODS: Subjects were all patients between 12-25 years of age treated at Pennsylvania trauma centers in 1996 who were reported to the state trauma database. Data on age, mechanism of injury (E-code), blood alcohol concentration (BAC), Injury Severity Score (ISS), and survival were obtained from the state database. BAC positive and negative patients within three groups: ages 12-17, 18-20, and 21-25, were compared to determine any difference in death rate, injury severity or type of injury. RESULTS: 4309 patients aged 12 25 were reported to the state in 1996. 2724 (63.2%) underwent testing for BAC, with 884 (32.5%) of those tested being positive. Testing positive were: 93/726 (12.8%) between 12-17 years old, 249/844 (29.5%) between 18-20 years old, and 542/1154 (47.0%) between 21-25 years old. 567/884 (64.1%) of those testing positive had BAC > or = 100 mg/dl. There were no statistically significant differences in mean ISS or death rate between BAC negative and BAC positive patients in either of the age groups. Regression analysis also showed no relationship between mortality and either the presence of alcohol or the actual level of BAC. Other regression analysis demonstrated a slight downward trend for ISS with increasing intoxication, which was statistically significant at P < 0.01. CONCLUSIONS: Alcohol ingestion is found even in early adolescent trauma patients and is seen to increase throughout the teenage years, occurring in over one-quarter of patients 18 to 20 years of age. Suspicion must be high that ingestion of alcohol has occurred in adolescent trauma. Further efforts should be made to improve the rate of testing in late adolescents, to ensure adequate identification of all alcohol-exposed patients and enable educational interventions. No significant differences in mortality were seen between alcohol positive and negative patients, but there was a trend to decreased injury severity with the presence of alcohol. PMID- 11063359 TI - Appropriateness of endotracheal tube size and insertion depth in children undergoing air medical transport. AB - OBJECTIVES: Guidelines for pediatric endotracheal tube (ETT) size and insertion depth are important in the helicopter EMS (HEMS) setting, where intubated patients are frequently transported by a non-physician flight crew providing protocol-based care in an environment noted for limitations in clinical airway assessment. The objectives of this study were to characterize, in a HEMS pediatric population, the frequency of compliance with guideline-recommended ETT size and insertion depth, and to test for association between guideline noncompliance and subsequent receiving hospital adjustment of ETT size or insertion depth. DESIGN: This retrospective review analyzed 216 consecutive pediatric (age <14) scene and interfacility HEMS transports, of patients intubated before or during HEMS transport, by an urban two-helicopter HEMS service providing protocol-based care with a nurse/paramedic crew configuration. Patients were transported to one of three receiving academic pediatric referral centers. Pediatric Advanced Life Support (PALS) criteria for ETT size and insertion depth were used to assess guideline-appropriateness of pediatric ETTs. Receiving hospital records were reviewed to determine if post-transport ETT size or lipline adjustment were associated with guideline-appropriateness of size and lipline during HEMS transport. Univariate (chi-square and Fisher's exact) and multivariate (logistic regression) statistics were used to assess and control for the following covariates: intubator group (physician, flight crew, ground EMS), transport year, sex, age, transport type (scene versus interfacility), and receiving hospital. For all analyses, statistical significance was set at the 0.05 level. RESULTS: The initial ETT size was within 0.5 mm of guideline recommended sizes in 178 (83.6%) of the 213 patients for whom this data were available. Inappropriate sized ETTs were nearly always (32 of 35, 91.4%) too small. Compared to initial ETTs placed by ground EMS personnel, initial ETTs placed by flight crew or physicians were more likely to be appropriate as defined by guidelines (P = .008 and .032, respectively). Receiving hospitals changed the ETT size in 18 (8.3% of 216) cases. Receiving hospital ETT size change was more likely with later transport year (P = .018) and less likely in patients over 2 years of age (P = .03); there was no significant association between receiving hospital ETT size change and intubator group (P > .22) or guideline appropriateness of ETT size (P = 0.94). The initial ETT insertion depth was within 1 cm of the guideline-recommended lipline in 86 (43.2%) of the 199 patients for whom this data were available. Inappropriate liplines were almost always (109 of 113, 96.5%) too deep. Compared to initial ETT liplines determined by ground EMS personnel, initial liplines determined by flight crew (P = .007), but not physician (P = .47) were more likely to be appropriate as defined by guidelines. Receiving hospitals changed the ETT insertion depth in 72 (33.3% of 216) cases. Receiving hospital lipline change was more likely (P = .03) in patients older than 2 years of age, but was not associated with intubator group (P = .75) or lipline guideline-appropriateness (P = .35). CONCLUSIONS: As judged by frequently used guidelines, pediatric ETTs are often too small and commonly inserted too deep. However, this retrospective study, limited by lack of clinical correlation for ETT size and insertion depth, failed to find an association between lack of ETT size or lipline guideline compliance and subsequent ETT adjustment at receiving pediatric centers. This study's findings, which should be confirmed with prospective investigation, cast doubt upon the utility of pediatric ETT size/lipline guidelines as strict clinical or quality assurance tools for use in pediatric airway management. PMID- 11063360 TI - Preliminary experience with 2-octylcyanoacrylate in a pediatric emergency department. AB - OBJECTIVE: Documentation of use of octylcyanoacrylate adhesives in a pediatric emergency department with reference to patient selection, complications, and parent satisfaction. DESIGN: Retrospective and concurrent chart review of the first 100 patients on which 2-octylcyanoacrylate (2-OCA, Dermabonda) was used in a pediatric emergency department. Additional telephone follow-up was performed for each patient. RESULTS: The average patient age was 4.7 years, average laceration size 1.2 cm. Sixteen percent of wounds were repaired with 2-OCA. Three immediate complications involved a minor dehiscence and two eyelid adhesions. Two wound infections and a patient with hematoma and keloid formation were identified as later complications. The vast majority of parents preferred tissue adhesive repair to sutures. Time in department was reduced from 106 minutes to 69 minutes on average (P < 0.0001, CI 26-52). CONCLUSIONS: Octylcyanoacrylate adhesives performed well in the daily practice of a pediatric emergency department, and were used for a significant percentage of laceration closures. Convenience, average infection rates, and good parental satisfaction make tissue adhesives a valuable addition to our wound closure techniques. Certain pitfalls occurring during early experience with these adhesives can be recognized and avoided. PMID- 11063361 TI - Transferring patients to a pediatric trauma center: the transferring hospital's perspective. AB - The rationale behind a regionalized trauma system is that patient outcomes are improved when trauma patients are rapidly transported to facilities with the level of expertise need to treat their injury. Functioning as an adult Level II trauma center, we wanted to know how the transfer process worked for pediatric patients whom we transfer to a Level I pediatric trauma center, which is part of the same multihospital system. Complete information on time of arrival, the time the transfer was accepted, and patient departure time were available for 116 patients (72% of pediatric patients transferred) for the period of January 1, 1997 through June 30, 1998. Patients were retrospectively stratified into two priority groups representing differing transport priority, based on use of a nasogastric tube, endotracheal tube or Foley catheter. Means for decision time and total time in transferring hospital were inspected. Decision time was 44 minutes (standard error 4.5 minutes) for priority patients and 92 minutes (11.5) for non-priority patients (t = 2.94, df = 114, P = 0.004). Total time for priority patients was 129 minutes (7.6) and 197 minutes (14.0) for non-priority patients (t = 3.37, df = 114, P = 0.001). Decision time was not influenced by extensive injury assessment or secondary studies. On average, pediatric patients spent nearly three hours in our facility. Our data indicate that a shorter decision time did not necessarily result in a reduction in wait time. Improving pediatric transfer times requires attention not only to injury assessment processes at the transferring facility and interhospital communications but also mobilization, hand-over, and any space or personnel constraints at the receiving pediatric facility. PMID- 11063362 TI - Oral versus intravenous: rehydration preferences of pediatric emergency medicine fellowship directors. AB - OBJECTIVE: The American Academy of Pediatrics (AAP) recommends oral rehydration therapy (ORT) for management of uncomplicated childhood gastroenteritis with mild moderate dehydration. However, ORT is widely underused relative to their recommendations. We compared ORT use by directors of Pediatric Emergency Medicine (PEM) fellowship training programs with AAP recommendations, and sought to identify their barriers to ORT. METHODS: Mail/fax survey of the directors of U.S. and Canadian PEM fellowship programs. The survey included 10 scenarios of mild or moderately dehydrated children with gastroenteritis, a personal innovativeness scale, self-assessment of ORT experience and knowledge, and open-ended questions regarding perceived barriers to ORT use. RESULTS: 60/67 (89.6%) PEM fellowship program directors responded. All reported experience with and knowledge about ORT. Only 10/58 (17.2%) believe ORT is usually better than intravenous (i.v.) rehydration in all 10 clinical scenarios, and only 4/58 (6.7%) usually use ORT in all 10 scenarios. 18/58 (31%) usually use ORT for all mildly but no moderately dehydrated children. ORT use did not correlate with personal innovativeness scores. Important barriers cited by respondents include additional time requirements for ORT relative to i.v. rehydration (76.7%) and expectation of i.v. rehydration by parents (41.7%) or primary care physicians (10%). CONCLUSIONS: Relative to AAP recommendations, PEM fellowship directors underuse ORT, especially for moderately dehydrated children. Physician innovativeness does not influence ORT use. Further study of effectiveness, length of stay, staff requirements, and ORT acceptance in the emergency department setting, especially in children with moderate dehydration, may influence ORT use. PMID- 11063363 TI - Dental floss ingestion requiring endoscopic retrieval. AB - We report an unusual case of a toddler who ingested dental floss and who subsequently was intubated in a community ED prior to transfer to our pediatric tertiary care center for endoscopic removal of the foreign body. PMID- 11063364 TI - Severe neurotoxic reaction associated with oral ingestion of low-dose diethyltoluamide-containing insect repellent in a child. AB - N,N-Diethyl-m-toluamide (DEET) is the major component of almost all tick repellent products. Reports of severe adverse reactions following voluntary ingestion are rare and primarily involve adults. This report describes a case of a toxic reaction after ingestion of low doses (80 mg/kg) of DEET in a child. The signs and symptoms were coma and seizures within 2 hours of ingestion. The patient recovered without sequelae. It is concluded that even scant doses of DEET may provoke severe encephalopathy in small children, and caution should be used when leaving this chemical unattended. PMID- 11063365 TI - Facial paralysis secondary to acute otitis media. AB - We describe a case of facial paralysis in a 19-month-old male recently diagnosed with acute otitis media. Results of his physical examination was remarkable for left-sided peripheral facial nerve palsy with an associated middle ear infection. Physicians should understand the etiology, pathophysiology, treatment options, and prognosis of facial palsy in association with otitis media. PMID- 11063366 TI - Inadvertent tarsorrhaphy secondary to Dermabond. PMID- 11063367 TI - Refractory priapism of unknown etiology in a pediatric patient. AB - Priapism is a urologic emergency that can occur in any age group and every patient should receive prompt urologic consultation. Management of priapism is based on the recognition of underlying pathophysiology; ready differentiation between high-flow and low-flow priapism; reversal of any potential precipitating factors; the use of corporal aspiration irrigation combined with intracavernosal alpha adrenergic therapy; and when necessary, a shunting procedure. Delay in recognition or treatment can be crucial as the incidence of long-term complication rises substantially with the duration of the priapism. PMID- 11063368 TI - Pyomyositis of the leg with early neurologic compromise. AB - Pyomyositis, although uncommon, is being reported with greater frequency in temperate climates. The presentation is similar to a number of infectious processes, and when associated with a traumatic event, the clinical picture may be confused with that of a musculoskeletal injury. This, coupled with an unfamiliarity of the disease, may result in a delay in diagnosis. Early antibiotic therapy may obviate surgery. Progression to the suppurative stage requires surgical drainage along with antibiotics. CT guided drainage may be accomplished in certain cases. In immunocompromised patients, progression to the septicemic stage is associated with high morbidity and mortality. PMID- 11063369 TI - The use of physical and chemical restraints in the pediatric emergency department. AB - Restraining patients is potentially dangerous and should be viewed as a last resort, to be used when no other modality of care is sufficient or when other efforts to calm the patient have been exhausted. Protocols and staff training are essential to limit inappropriate use of restraints and to protect both the patient and staff. Further clinical studies are needed in the area of chemical restraint of children and to evaluate the safety and efficacy of different methods of physical restraint. PMID- 11063370 TI - Case record of the Children's Mercy Hospital: a 15 year old with hoarseness following neck trauma. PMID- 11063371 TI - Trials and tribulations of the asthmatic patient. PMID- 11063372 TI - Pediatric emergency medicine: legal briefs. PMID- 11063373 TI - Serial bedside emergency ultrasound in a case of pediatric blunt abdominal trauma with severe abdominal pain. AB - We present a case of a teenager with isolated left renal laceration with perirenal hematoma. The patient had presented with severe left upper quadrant (LUQ) pain following blunt abdominal trauma (BAT) sustained during a sledding accident. A screening bedside focused abdominal sonogram for trauma (FAST) rapidly excluded free fluid on two serial examinations, 30 minutes apart. It provided the pediatric emergency physician with a measure of diagnostic confidence that the patient could be safely transported to the CT suite for detailed delineation of his injury. Moreover, narcotic analgesia was liberally administered early in his illness course, without the fear of unmasking potential hypovolemia when it was known that he did not have gross intra-abdominal bleeding on his bedside ultrasound (US). It also provided a working diagnosis of the primary organ of injury. Our hospital, like many pediatric hospitals around the nation, does not have in-house 24-hour radiology support. We suggest that the use of the bedside US be extended to the stable pediatric patient in severe abdominal pain following BAT. It can serve as a valuable, rapid, noninvasive, bedside, easily repeated, fairly accurate triage tool to evaluate pediatric BAT with severe pain. PMID- 11063374 TI - Methylenedioxymethamphetamine (MDMA, "Ecstasy"). AB - Ecstasy (MDMA) is an amphetamine derivative of growing popularity. The drug produces a range of toxicities when taken either in standard doses or overdose. In overdose it has major toxicity, producing several different life-threatening manifestations. Hepatotoxicity and hyponatremia are common but poorly understood consequences of MDMA overdose. The drug can produce long-term, if not permanent, neurologic sequelae by destruction of serotonergic neurons. Chronic Ecstasy use can result in psychosis, depression, and suicidal ideation. In the ED setting, it is essential for physicians to recognize and treat appropriately those who present with intoxication from this drug. PMID- 11063375 TI - Assault with bruising and drainage from eye. PMID- 11063376 TI - High-dose chemotherapy in breast cancer: the end of the beginning? PMID- 11063377 TI - Status of high-dose chemotherapy for breast cancer: a review. AB - The purpose of this review is to analyze the current status of high-dose chemotherapy (HDCT) with autologous stem cell transplantation for patients with breast cancer. Current results from the major prospective phase 2 and phase 3 trials in metastatic breast cancer (MBC) and high-risk primary breast cancer (HRPBC) are reviewed. Prognostic factors and future research directions are also discussed. The encouraging results of phase 2 trials suggested a benefit for HDCT in HRPBC and some categories of patients with MBC. Some investigators have argued that patient selection might have been a critical factor in those studies. Recently reported randomized trials in patients with chemosensitive MBC have included only small numbers of patients in complete remission and thus have not adequately addressed the relative value of HDCT versus maintenance standard-dose chemotherapy in this patient subset. Although initial results of 2 studies have been reported, most randomized phase 3 studies of HDCT in HRPBC require longer follow-up before definitive conclusions can be made about its efficacy in this setting. We conclude that the role of HDCT for HRPBC or MBC patients has not yet been fully defined. Longer follow-up of the ongoing randomized trials is necessary, and their mature results will help clarify this important question. In the meantime, it is imperative that research continues, to enhance the efficacy of the procedure. This may come through incorporating more active drugs into HDCT regimens and combining HDCT with novel strategies aimed at eradication of posttransplantation minimal residual disease. PMID- 11063378 TI - High-dose chemotherapy and hematopoietic stem cell rescue for breast cancer: experience in California. AB - The role of high-dose chemotherapy (HDCT) and autologous hematopoietic stem cell rescue in breast cancer is still controversial. We analyzed the outcomes of 1111 consecutive patients with histologically proven breast cancer who underwent HDCT at 5 major California medical centers. The overall treatment-related mortality (TRM) was 2.3%. TRM was not influenced by disease stage or the HDCT regimen delivered, but it was influenced by hematopoietic graft source. The TRM was 6.1% when bone marrow with or without blood stem cells was used, but only 1.4% when blood stem cells alone were used (P < .001). With a median follow-up of 2.8 years (range, 0.1-8.2 years) after HDCT and autologous hematopoietic stem cell rescue, the estimated 5-year event-free survival (EFS) and overall survival (OS) for stage II/IIIA patients with > or =10 involved axillary lymph nodes were 67% and 76%, respectively. Patients with metastatic breast cancer (MBC) (median follow up, 1.9 years [range, 0.03-8.3 years]) achieving a complete response (CR) to conventional-dose chemotherapy or rendered to a "no evidence of disease" status before HDCT had significantly better estimated 5-year EFS and OS (28% and 57%, respectively) than those achieving a partial response before HDCT (19% and 27%, respectively; P < or = .0001). Our data suggest that HDCT with hematopoietic stem cell rescue is safe and can be beneficial to patients with high-risk primary breast cancer and for those with MBC achieving CR/no evidence of disease. PMID- 11063379 TI - Favorable treatment outcome in non-Hodgkin's lymphoma patients with "poor" mobilization of peripheral blood progenitor cells. AB - Our purpose was to evaluate the outcome and costs of high-dose chemotherapy and autologous peripheral blood progenitor cell (PBPC) transplantation in patients with the inability to mobilize sufficient numbers of PBPCs to allow rapid engraftment after PBPC transplantation. We treated 172 consecutive non-Hodgkin's lymphoma (NHL) patients with cyclophosphamide and granulocyte colony-stimulating factor followed by apheresis to collect PBPCs. The cells were separated on a Percoll gradient and purged with monoclonal antibodies and complement. The patients were categorized as "good" mobilizers if a collection of > or =2 x 10(6) CD34+ cells/kg was obtained (n = 138, 80%) or "poor" mobilizers if <2 x 10(6) CD34+ cells/kg were obtained (n = 34, 20%). With a median follow-up of 3.5 years, there is no statistically significant difference in actuarial event-free survival, overall survival, or relapse for good mobilizers compared with poor mobilizers. However, there was a trend toward increasing nonrelapse, transplantation-related mortality of 11.8% for poor mobilizers versus 3.6% for good mobilizers (P = .08) and early death from all causes including relapse within 120 days (poor 20.6% versus good 8.7%, P = .06). The total cost for bone marrow transplantation-related care was significantly higher, at $140,264 for poor mobilizers versus $80,833 for good mobilizers (P = .0001). The population of patients with NHL who mobilize PBPCs poorly into the circulation have a higher cost for posttransplant support. However, there is no significant difference in relapse, event-free survival, or overall survival for such patients compared with those who mobilize PBPCs easily. PMID- 11063380 TI - Prevention of coronary vascular disease by transplantation of T-cell-depleted bone marrow and hematopoietic stem cell preparation in autoimmune-prone w/BF(1) mice. AB - This project was designed to investigate the application of bone marrow transplantation to a progressive and ultimately fatal systemic autoimmune disease. Male (NZW x BXSB)F1 (W/BF1) mice develop acute systemic autoimmune disease characterized by degenerative coronary vascular disease (CVD) with myocardial infarctions, hypertension, thrombocytopenia, glomerulonephritis, and persistently elevated levels of circulating immune complexes. After preliminary studies established the onset of disease between 10 and 12 weeks of age, 6- to 8 week-old male W/BF1 mice were targeted for transplantation with either T-cell depleted bone marrow or purified hematopoietic stem cells from haploidentical B6C3/F1 mice. Posttransplantation flow cytometric analysis of splenocytes demonstrated donor phenotypes in W/BF1 recipient mice that had received T-cell depleted marrow or hematopoietic stem cell preparations (lineage negative, CD71 negative) from B6C3/F1 donors. Survival of W/BF1 mice transplanted with bone marrow from normal B6C3/F1 donors was very high, and assessment at 100 days after transplantation revealed reduction in onset and severity of disease. Autoantibodies to cardiolipin and double-stranded DNA were markedly reduced to levels present in normal mice. Immunohistochemistry of heart and kidney tissue revealed significant amelioration of degenerative CVD and glomerulonephritis in the majority of W/BF1 recipients of marrow transplants from B6C3/F1 donors. All engrafted W/BF1 mice displayed normal immunologic competence 100 days posttransplantation. PMID- 11063381 TI - Evidence-based reviews and the role of blood and marrow transplantation in the treatment of selected disease: an ASBMT policy statement. American Society for Blood and Marrow Transplantation. PMID- 11063382 TI - ASBMT policy statement regarding the methodology of evidence-based reviews in evaluating the role of blood and marrow transplantation in the treatment of selected diseases. American Society for Blood and Marrow Transplantation. PMID- 11063383 TI - Relationship of blood glucose level to community periodontal index of treatment needs and body mass index in a permanent Israeli military population. AB - BACKGROUND: The goal of this study was to find a possible link between blood glucose levels and periodontal disease. METHODS: In a prospective study the dental health of 10,590 military service men and women was examined. The relationships of periodontal condition, expressed as CPITN index, and blood glucose levels were tested. The effect of gender, body mass index (BMI), and smoking was also evaluated. RESULTS: Blood glucose levels were significantly and positively associated with severe periodontal disease, men, and higher BMI. Smoking did not affect blood glucose levels. CONCLUSIONS: Blood glucose levels might be associated with severe periodontal disease. PMID- 11063384 TI - Elevation of systemic markers related to cardiovascular diseases in the peripheral blood of periodontitis patients. AB - BACKGROUND: Periodontitis is a common, often undiagnosed, chronic infection of the supporting tissues of the teeth, epidemiologically associated with cardiovascular diseases. Since C-reactive protein (CRP) and other systemic markers of inflammation have been identified as risk factors for cardiovascular diseases, we investigated whether these factors were elevated in periodontitis. METHODS: Consecutive adult patients with periodontitis (localized n = 53; generalized n = 54), and healthy controls (n = 43), all without any other medical disorder, were recruited and peripheral blood samples were taken. RESULTS: Patients with generalized periodontitis and localized periodontitis had higher median CRP levels than controls (1.45 and 1.30 versus 0.90 mg/L, respectively, P = 0.030); 52% of generalized periodontitis patients and 36% of the localized periodontitis patients were sero-positive for interleukin-6 (IL-6), compared to 26% of controls (P= 0.008). Plasma IL-6 levels were higher in periodontitis patients than in controls (P = 0.015). Leukocytes were also elevated in generalized periodontitis (7.0 x 10(9)/L) compared to localized periodontitis and controls (6.0 and 5.8 x 10(9)/L, respectively, P= 0.002); this finding was primarily explained by higher numbers of neutrophils in periodontitis (P= 0.001). IL-6 and CRP correlated with each other, and both CRP and IL-6 levels correlated with neutrophils. The current findings for periodontitis were controlled for other known factors associated with cardiovascular diseases, including age, education, body mass index, smoking, hypertension, cholesterol, and sero positivity for CMV, Chlamydia pneumoniae, and Helicobacter pylori. CONCLUSIONS: Periodontitis results in higher systemic levels of CRP, IL-6, and neutrophils. These elevated inflammatory factors may increase inflammatory activity in atherosclerotic lesions, potentially increasing the risk for cardiac or cerebrovascular events. PMID- 11063385 TI - Levels of interleukin-1 beta, -8, and -10 and RANTES in gingival crevicular fluid and cell populations in adult periodontitis patients and the effect of periodontal treatment. AB - BACKGROUND: Various cytokines have been identified at sites of chronic inflammation such as periodontitis. Cytokines are synthesized in response to bacteria and their products, inducing and maintaining an inflammatory response in the periodontium. The purpose of the present study was to investigate the involvement of interleukin-1 beta (IL-1 beta), IL-8, and IL-10 and RANTES (regulated on activation, normally T cell expressed and secreted) and the cell populations associated with the immune response in destructive periodontitis, as well as the effect of periodontal therapy on cytokine levels in gingival crevicular fluid (GCF). METHODS: Data were obtained from 12 patients with moderate to advanced periodontitis and 6 healthy controls. Patients presenting at least 2 sites with > or =2 mm clinical attachment loss were included in the destructive periodontitis group. After monitoring for 4 months, only 6 patients showed destructive periodontitis and GCF samples and soft tissues biopsies were collected from these patients. GCF samples and biopsies were collected both from active (12 CGF samples and 6 biopsies) and inactive (12 CGF samples and 6 biopsies) sites. The comparison with healthy controls was carried out by collecting GCF samples from 6 healthy volunteers (12 samples) and biopsies during the surgical removal of wisdom teeth. In periodontal patients, clinical data and GCF samples were obtained prior to periodontal treatment (72 samples) and 2 months after periodontal therapy (72 samples). GCF was collected using a paper strip; eluted and enzyme-linked immunoabsorbent assays (ELISA) were performed to determine cytokine levels. The inflammatory infiltrate was analyzed by immunohistochemistry of gingival biopsy samples with monoclonal antibodies against CD3, CD8, CD4, CD11c, and CD19 antigens. RESULTS: Cellular components of the inflammatory infiltrate include B and T lymphocytes and monocyte/macrophages. Active sites contained a higher number of B lymphocytes and macrophages. IL-8 and IL-1 beta and RANTES in GCF were detected in the majority of sites from periodontal patients (100%, 94% and 87%, respectively); IL-10 was found in only 43%. IL-8 was the only cytokine detected in the GCF (75%) of the control group. Moreover, IL-1 beta levels were significantly higher in active sites versus inactive sites (P <0.05). IL-8 and IL-10 and RANTES were increased in active sites; however, differences were not significant (P>0.05). A positive correlation between the IL-8 and RANTES (r = 0.677, P<0.05) was observed in periodontitis patients. Periodontal therapy reduced the total amount of IL-1 beta, IL-8, and IL 10 and RANTES. Data showed a weak correlation between the clinical parameters and the total amount of cytokines in periodontitis. CONCLUSIONS: These data suggest that the amount of crevicular IL-1 beta, IL-8, and IL-10 and RANTES is associated with periodontal status. Removal of the bacterial plaque reduces the antigenic stimuli and consequently could modulate the chemokines present in GCF. We propose that the dynamic interactions between cytokines, their production rates, and their quantity could represent factors controlling the induction, perpetuation, and collapse of the cytokine network present in the periodontal disease. PMID- 11063386 TI - Effects of roxithromycin on tumor necrosis factor-alpha-induced vascular endothelial growth factor expression in human periodontal ligament cells in culture. AB - BACKGROUND: Aberrant angiogenesis is associated with lesion formation in chronic periodontitis. However, little is known about the mediators that contribute to angiogenesis or about therapeutic agents that control the production of the mediators. Roxithromycin (RXM), which is a new 14-member macrolide antibiotic, has a wide antibacterial spectrum against oral pathogens and an immunomodulatory effect. In the present study, we examined the effects of RXM on tumor necrosis factor (TNF)-alpha-induced vascular endothelial growth factor (VEGF) in human periodontal ligament (HPDL) cells. In addition, the effect of RXM on VEGF expression in HPDL cells was examined. METHODS: HPDL cells were plated at 5 x 10(5) cells/ml in 150 cm2 cell culture dishes. The confluent-stage cells were pretreated with or without 10 microg/ml of RXM or other antibiotics in 1% FBS containing alpha-MEM for 24 hours, followed by simultaneous treatment with 10 ng/ml of TNF-alpha and 10 microg/ml of these antibiotics. After incubation for various periods, the culture supernatants and sediments were collected and analyzed by ELISA, Northern blot, and gel shift assays. RESULTS: VEGF mRNA and its protein were constitutively expressed in HPDL cells, and the level of expression was markedly enhanced by stimulation with TNF-alpha. RXM strongly inhibited the expression of VEGF mRNA and the production of VEGF. Furthermore, RXM suppressed activation of transcription factors AP-1 and SP-1, which were critical factors in VEGF transcription, in TNF-alpha-stimulated HPDL cells. CONCLUSION: These results indicate that TNF-alpha, one of the proinflammatory cytokines implicated in the pathogenesis of periodontitis, induces excess induction of VEGF in HPDL, which may account for increased angiogenesis in periodontitis lesions. Interestingly, the antibiotic roxithromycin inhibits TNF mediated VEGF induction, suggesting its possible therapeutic utility in periodontitis and other chronic inflammatory conditions involving VEGF induction. PMID- 11063387 TI - Identification of periodontal pathogens in atheromatous plaques. AB - BACKGROUND: Recent studies suggest that chronic infections including those associated with periodontitis increase the risk for coronary vascular disease (CVD) and stroke. We hypothesize that oral microorganisms including periodontal bacterial pathogens enter the blood stream during transient bacteremias where they may play a role in the development and progression of atherosclerosis leading to CVD. METHODS: To test this hypothesis, 50 human specimens obtained during carotid endarterectomy were examined for the presence of Chlamydia pneumoniae, human cytomegalovirus, and bacterial 16S ribosomal RNA using specific oligonucleotide primers in polymerase chain reaction (PCR) assays. Approximately 100 ng of chromosomal DNA was extracted from each specimen and then amplified using standard conditions (30 cycles of 30 seconds at 95 degrees C, 30 seconds at 55 degrees C, and 30 seconds at 72 degrees C). Bacterial 16S rDNA was amplified using 2 synthetic oligonucleotide primers specific for eubacteria. The PCR product generated with the eubacterial primers was transferred to a charged nylon membrane and probed with digoxigenin-labeled synthetic oligonucleotides specific for Actinobacillus actinomycetemcomitans, Bacteroides forsythus, Porphyromonas gingivalis, and Prevotella intermedia. RESULTS: Eighty percent of the 50 endarterectomy specimens were positive in 1 or more of the PCR assays. Thirty eight percent were positive for HCMV and 18% percent were positive for C. pneumoniae. PCR assays for bacterial 16S rDNA also indicated the presence of bacteria in 72% of the surgical specimens. Subsequent hybridization of the bacterial 16S rDNA positive specimens with species-specific oligonucleotide probes revealed that 44% of the 50 atheromas were positive for at least one of the target periodontal pathogens. Thirty percent of the surgical specimens were positive for B. forsythus, 26% were positive for P. gingivalis, 18% were positive for A. actinomycetemcomitans, and 14% were positive for P. intermedia. In the surgical specimens positive for periodontal pathogens, more than 1 species was most often detected. Thirteen (59%) of the 22 periodontal pathogen-positive surgical specimens were positive for 2 or more of the target species. CONCLUSIONS: Periodontal pathogens are present in atherosclerotic plaques where, like other infectious microorganisms such as C. pneumoniae, they may play a role in the development and progression of atherosclerosis leading to coronary vascular disease and other clinical sequelae. PMID- 11063388 TI - Some effects of non-surgical therapy on gingival inflammatory cell subsets in patients with adult and early-onset periodontitis. AB - BACKGROUND: Limited information is available to determine if there is a distinction in local cellular immunity between adult and early-onset periodontitis. Furthermore, the effect of scaling and root planing on various lymphocyte subsets is sparsely described. METHODS: Clinical measurements were recorded and gingival biopsies were performed before and after scaling and root planing in 10 subjects with adult (AP) and in 10 with early-onset periodontitis (EOP). The specimens were cut into serial sections and, using the alkaline phosphatase-anti-alkaline phosphatase technique, monoclonal antibodies to CD20 (B cells), CD30 (plasma cells), and CD45RO (T-memory cells) were applied, as well as polyclonal antibodies to alpha, gamma, and mu chains (Ig A, G, and M). Areas showing the largest infiltration cells were counted. RESULTS: Mean counts of all cell phenotypes in the AP versus the EOP group did not show any significant differences before therapy (P >0.05). Following scaling and root planing, numbers of all phenotypes decreased in both groups. Comparing the data before and after therapy, P values were >0.05 in the AP group, except for IgA-positive cells. In the EOP group, the differences before and after therapy reached statistical significance (P<0.05) for all cell counts, except for IgM-positive cells. Furthermore, reduction of probing depth and gain of clinical attachment reached the 0.05 level of statistical significance only in EOP subjects. CONCLUSIONS: Local cellular immunity in patients with adult onset periodontitis does not appear to differ from the immune response in patients with early onset periodontitis. Scaling and root planing causes a decrease in the inflammatory cells subsets tested, however, this decline seems to be more pronounced in EOP than in AP subjects. PMID- 11063389 TI - The effect of age on proliferating cell nuclear antigen expression in oral gingival epithelium of healthy and inflamed human gingiva. AB - BACKGROUND: Age-related changes in proliferative activity in human gingival epithelium are uncertain. Proliferating cell nuclear antigen (PCNA) is a nuclear protein associated with the cell cycle. Nuclear PCNA immunoreactivity is found in the proliferative compartment of normal tissues. The aims of this study were to investigate the localization of PCNA expression in oral gingival epithelium (OGE) and to define the age-related changes as to PCNA-proliferative index (PI) in inflamed as well as healthy gingiva. Mitotic index (MI) was also used as a conventional marker of cell proliferation. Additionally, the effect of aging upon the maximum epithelial thickness (MET) was determined. METHODS: Twenty older (65 to 85 years) (study) and 20 middle-aged (35 to 45 years) (controls) subjects were included in the study. Biopsies were obtained both from healthy and inflamed gingiva. The expression of PCNA was evaluated in formalin-fixed, paraffin embedded gingival samples using an immunoperoxidase technique and PC 10 monoclonal antibody to PCNA. Hematoxylin and eosin stained sections were used for the quantitative measurement of MI and MET. RESULTS: All the tissue sections contained positive staining cells for PCNA in the gingival epithelium. Although PCNA expression was observed both in the basal and suprabasal layers, it was more prominent in the suprabasal layers. PI in inflamed gingiva was significantly higher in the older group. However, no significant difference was observed between the study and control groups with respect to PI in healthy gingiva. When all the subjects taken into the study were analyzed as a single group, PI in the inflamed gingival samples were found to be increased with aging. Nevertheless, no age-related change was noted in MI and MET. In both the study and the control groups, PI, MI, and MET were found to be increased due to inflammation. CONCLUSIONS: Our data indicate that PCNA expression in inflamed gingiva is higher in older subjects. Furthermore, a significant correlation was noted between aging and PCNA expression in inflamed gingiva. As there is no increase in mucosal epithelial thickness despite increased proliferation, we speculate that the duration of the PCNA+ phase in cell cycle may be longer in older subjects. This study also implies that PCNA immunolocalization can be used as an index of the state of cell proliferation in both biological and pathological events of the gingiva and/or other mucosal tissues. PMID- 11063390 TI - Cyclooxygenase-2-dependent prostaglandin production by peripheral blood monocytes stimulated with lipopolysaccharides isolated from periodontopathogenic bacteria. AB - BACKGROUND: Prostaglandin E2 (PGE2) plays important roles in the pathogenesis of periodontal disease. Recent studies have revealed the existence of 2 isozymes of cyclooxygenase (COX), called COX-1 and COX-2. The purpose of the present study was to investigate the contribution of COX-1 and COX-2 to PGE2 production by human peripheral blood monocytes that are stimulated with lipopolysaccharides (LPS) from periodontopathogenic bacteria. METHODS: LPS were isolated from Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) and Porphyromonas gingivalis (P. gingivalis) by the phenol-water method. Peripheral blood monocytes were stimulated with LPS for the indicated periods, and the levels of PGE2 or interleukin (IL)-1 beta in the culture media were measured by enzyme-linked immunosorbent assay. Expression of COX-1 and -2 proteins was studied by immunocytochemical staining, and COX-2 mRNA expression was examined by Northern blot analysis. RESULTS: Peripheral blood monocytes stimulated with A. actinomycetemcomitans- or P. gingivalis-LPS produced PGE2 in a time- and dose dependent manner. Indomethacin, a non-selective COX-1/COX-2 inhibitor, and NS 398, a specific COX-2 inhibitor, completely inhibited PGE2 production. Immunocytochemical staining of COX-1 and COX-2 proteins showed that expression of COX-2 protein was increased in monocytes that were stimulated with A. actinomycetemcomitans- or P. gingivalis-LPS, compared with that in unstimulated monocytes, whereas expression of COX-1 protein was not altered. Northern blot analysis showed that monocytes stimulated with A. actinomycetemcomitans- or P. gingivalis-LPS expressed COX-2 mRNA, while COX-2 mRNA was not detectable in unstimulated cells. Treatment of A. actinomycetemcomitans-LPS-stimulated monocytes with NS-398 induced a significant increase of IL-1 beta production to the same extent as treatment with indomethacin. CONCLUSIONS: These results suggest that COX-2 is induced in monocytes stimulated with LPS derived from A. actinomycetemcomitans and P. gingivalis and that the COX-2 is primarily responsible for PGE2 production. COX-2 may be pivotal in PGE2 production in periodontal lesions and may be involved in inflammatory responses. PMID- 11063391 TI - Periodontal disease progression. AB - BACKGROUND: The objective of this investigation is to use noninvasive, state-of the-art, diagnostic techniques to measure periodontal disease progression and model periodontal disease activity over time. In this investigation, digital subtraction radiography and an electronic controlled force periodontal probe capable of attachment level measurement were used to measure bone loss and attachment loss, respectively. The use of these nearly continuous measures of attachment and bone loss allowed detection of small amounts of disease activity and provided data to be used in modeling of the disease process over time. METHODS: Forty-four patients were studied for 18 months. Examinations used clinical attachment level measures at 1-month intervals and quantitative radiology at 6-month intervals. The sites were analyzed by regression for statistically significant changes. These data were used to determine sites of periodontal disease activity for testing various models of periodontal disease progression. RESULTS: Overall 22.8% of sites lost attachment, 5.4% gained, and 71.7% demonstrated no statistically significant change. The mean time to lose 1 mm of attachment was 8.4 +/- 0.6 months. In the first model tested a step-wise discriminant analysis was used to determine whether or not baseline measurements of plaque (PI), gingival inflammation (GI), attachment loss, and probing depth (PD) could be used to derive a satisfactory model for disease progression. Although the overall model was statistically significant with PI, PD, and GI contributing to the model (Wilks' lambda = 0.859, F = 5.71, P <0.0012), its predictive power was relatively weak. A considerably stronger significant model resulted when the rate of attachment loss over the first 6 months, baseline PI, and baseline GI were included (Wilks' lambda = 0.712, F = 14.17, P<0.00001). A significant model also resulted when bone loss during the first 6 months and baseline probing depth were included (Wilks' lambda = 0.438, F = 61.48, P<0.00001). When the last model was applied to each site, the sensitivity in predicting disease progression was 80.0% and the specificity in ruling out progressive disease was 93.9%. CONCLUSIONS: This study indicates that clinically significant progression of attachment loss in posterior tooth sites occurs as a frequent event in adult periodontitis. The modeling data also suggest that short term (6 month) measures of periodontal disease progression greatly improve the ability to model attachment loss over a longer period in untreated periodontitis patients. PMID- 11063392 TI - Growth factors regulate expression of mineral associated genes in cementoblasts. AB - BACKGROUND: Knowledge of the responsiveness of cells within the periodontal region to specific bioactive agents is important for improving regenerative therapies. The aim of this study was to determine the effect of specific growth factors, insulin-like growth factor-I (IGF-I), platelet-derived growth factor-BB (PDGF-BB), and transforming growth factor-beta (TGF-beta) on cementoblasts in vitro and ex vivo. METHODS: Osteocalcin (OC) promoter driven SV40 transgenic mice were used to obtain immortalized cementoblasts. Growth factor effects on DNA synthesis were assayed by [3H]-thymidine incorporation. Northern analysis was used to determine the effects of growth factors on gene expression profile. Effects of growth factors on cementoblast induced biomineralization were determined in vitro (von Kossa stain) and ex vivo (re-implantation of cells in immunodeficient (SCID) mice). RESULTS: All growth factors stimulated DNA synthesis compared to control. Twenty-four hour exposure of cells to PDGF-BB or TGF-beta resulted in a decrease in bone sialoprotein (BSP) and osteocalcin (OCN) mRNAs while PDGF-BB also increased osteopontin (OPN) mRNA. Cells exposed to IGF-I for 24 hours exhibited decreased transcripts for OCN and OPN with an upregulation of BSP mRNA noted at 72 hours. In vitro mineralization was inhibited by continuous application of PDGF-BB or TGF-beta, while cells exposed to these factors prior to implantation into SCID mice still promoted biomineralization. CONCLUSIONS: These data indicate IGF-I, PDGF-BB, and TGF-beta influence mitogenesis, phenotypic gene expression profile, and biomineralization potential of cementoblasts suggesting that such factors alone or in combination with other agents may provide trigger factors required for regenerating periodontal tissues. PMID- 11063393 TI - Amine fluoride/stannous fluoride and chlorhexidine mouthwashes as adjuncts to surgical periodontal therapy: a comparative study. AB - BACKGROUND: Postsurgical mouthwashes are routinely used in clinical studies and also in daily clinical practice. Chlorhexidine gluconate (CHX) has long been the gold standard for supra-gingival chemical plaque control regimens. Amine fluoride/stannous fluoride (AmF/SnF2) formulations have also been extensively studied and shown to have an antibacterial effect and be useful as antiplaque agents. The antibacterial effect of AmF/SnF2 and its minimal extrinsic tooth staining make it a possible alternative to CHX as an adjunct to periodontal surgical therapy. The aim of this double-blind, controlled clinical trial was to evaluate and compare the combined effect of an AmF/SnF2 or a CHX mouthwash and surgical periodontal therapy on periodontal parameters. METHODS: Thirty-two patients with at least 3 pockets > or =5 mm in the same quadrant were selected for this study, following a hygienic phase of therapy. They were randomized into 2 treatment groups: surgical flap debridement and a postsurgical CHX mouthwash or surgical flap debridement and an AmF/SnF2 postsurgical mouthwash, performed twice daily for 3 weeks. Clinical measurements were taken at baseline and 3 and 12 weeks postsurgery. RESULTS: Both treatment modalities resulted in significant improvements in probing depth and clinical attachment level. There was no significant difference between groups in any of the recorded parameters. Staining index at week 3 in the CHX group was significantly higher than in the AmF/SnF2 group (P<0.05). However these differences leveled down at 12 weeks. CONCLUSIONS: Our results support the alternative use of an AmF/SnF2 mouthwash in plaque control management of patients following flap debridement surgery. PMID- 11063394 TI - The effect of topical delivery of novel bisacylphosphonates in reducing alveolar bone loss in the rat model. AB - BACKGROUND: Periodontal surgery stimulates osteoclast activity, leading to varying amounts of alveolar crest loss. We have established that topical application of 20 mg/ml of alendronate placed at the surgical mucoperiosteal site produced a striking reduction of alveolar bone loss in the rat model. The aim of this investigation was to examine the antiresorptive efficacy of 3 novel bisacylphosphonates topically delivered at the surgical site, in comparison to alendronate and etidronate which are in clinical use. METHODS: Mucoperiosteal flap (MF) surgery was performed on the buccal and lingual aspects next to molars on both sides of the rat mandible. A gelatin sponge soaked in the bisphosphonate solution prepared by dissolving 20 mg of the bisphosphonate (alendronate, etidronate, VS-5, VS-6, ISA-13, SuBP) in 1 ml of saline was applied to exposed bone on the right side of the mandible (experimental, MF + BPs ) and the left side was treated with saline only (control, MF + S). Sections were evaluated for bone loss using microradiography pattern and amount. RESULTS: The 3 novel bisacylphosphonates, VS-5 VS-6, and ISA-13 were more effective than etidronate, and less effective than alendronate. The most effective among this group was ISA 13 followed by VS-5 and VS-6. CONCLUSION: We conclude that ISA-13-like alendronate is effective in reducing alveolar bone loss when delivered at surgical sites. Since ISA-13 is well absorbed through mucose tissues, we suggest that ISA-13 efficacy on reducing bone loss should be tested by its application on the mucosal tissue. PMID- 11063395 TI - Osteotome sinus floor elevation and simultaneous, non-submerged implant placement: case report and literature review. AB - The osteotome sinus floor elevation is a conservative technique for sinus elevation and immediate implant placement. The purpose of this report is to present a case using the sinus floor elevation technique for placement of a non submerged implant with 12 months follow-up after permanent restoration and to review the literature regarding sinus floor elevation. The osteotome sinus floor elevation technique may be used successfully for the placement of nonsubmerged implants. It allows for shorter post-treatment waiting times and provides a less invasive approach compared to 1- or 2-step antrostomy procedures. PMID- 11063396 TI - Combined treatment of periodontal disease and benign mucous membrane pemphigoid. Case report with 8 years maintenance. AB - BACKGROUND: Desquamative gingivitis may be the clinical manifestation of one of several systemic diseases. The clinical course of the disease can be complicated by plaque-associated periodontitis. However, there is no information currently available for the concurrent management of both conditions. CASE REPORT AND RESULTS: This paper presents the treatment and 8-year maintenance of a patient with periodontal disease and benign mucous membrane pemphigoid (BMMP). The first phase of treatment included oral hygiene instructions and local corticosteroid administration, followed by scaling and root planing. The patient's compliance and excellent response to therapy allowed for subsequent surgical pocket elimination and augmentation of the zone of keratinized tissue for prosthetic reasons. Over the following 8 years, the patient's periodontal condition remained stable even though periodontal maintenance was erratic. For the control of BMMP, intermittent administration of corticosteroids was necessary, without any significant local or systemic side effects. CONCLUSIONS: We suggest that combined treatment and long-term maintenance of BMMP and periodontitis are feasible under certain conditions and propose a clinical protocol for treatment which could serve as a guideline for similar conditions. PMID- 11063397 TI - Reduction of severe gingival overgrowth in a kidney transplant patient by replacing cyclosporin A with tacrolimus. AB - Side effects of certain drugs such as cyclosporin A (CsA) and phenytoin may induce gingival overgrowth which in some instances become unacceptable to the patient because esthetic, functional, and other effects. Use of these drugs is related to important medical situations, such as organ transplantation and control and withdrawal of the drugs is contraindicated. Tacrolimus is an immunosuppressant used to prevent graft rejection in organ transplant patients and has been shown to cause fewer oral side effects than CsA. This report deals with a case of probable synergism between the use of CsA and phenytoin which caused an intense gingival overgrowth in a kidney transplant patient. A treatment protocol including very thorough oral hygiene, scaling and root planing, clorhexidine digluconate rinses (0.12%), and substituting CsA with tacrolimus is described. Response to treatment after 6 months of tacrolimus use was excellent with almost complete reversion of the gingival enlargement. One-year follow-up demonstrated a stable gingival situation. The successful substitution of CsA with tacrolimus provides great expectations for the management of CsA-related gingival enlargement. PMID- 11063398 TI - Repair of an intrabony defect from an adenomatoid odontogenic tumor. AB - This case report describes the occurrence and treatment of an adenomatoid odontogenic tumor (AOT) presenting as a periodontal intrabony defect on a upper lateral incisor. Following incision and flap reflection, a solid, rubbery specimen was enucleated in one piece leaving a wide moat-like intrabony defect. A bioabsorbable membrane of glycolide and lactide copolymer was sutured over the defect to maximize regeneration and the flaps then sutured over the membrane. Microscopic analysis of the biopsied specimen revealed dental hard tissue interspersed in a field of odontogenic epithelium in a glandular configuration, consistent with a diagnosis of AOT. AOT is a rare odontogenic tumor usually associated with unerupted teeth or dentigerous cysts, not periodontal defects. Clinical evaluation 6 months postoperatively revealed restoration of clinical attachment and periodontal health. PMID- 11063399 TI - The use of a bioabsorbable barrier for regenerative management of marginal tissue recession. I. Report of 100 consecutively treated teeth. AB - BACKGROUND: Periodontal plastic surgical procedures aimed at coverage of exposed root surfaces have evolved into routine treatment modalities. The present study was designed to assess the effectiveness and the predictability of a bioabsorbable barrier in the treatment of human recession defects utilizing a single-step surgical procedure. METHODS: One hundred consecutive single and multiple adjacent Miller Class I, II, and III buccal recession defects in 41 patients were treated with a combination of a bioabsorbable barrier and coronally advanced flap technique. Clinical parameters were recorded immediately prior to surgery, at 3 months, and after a minimum of 6 months. RESULTS: A highly significant reduction in recession depth from a mean value of 3.2 +/- 0.9 mm preoperatively to 0.3 +/- 0.5 mm postoperatively, corresponding to a mean root coverage of 92. 7% +/- 14.1%, was obtained. Complete (100%) root coverage was obtained in 75% of the sites. Factors adversely affecting root coverage were membrane exposure postoperatively and preoperative recession depth > or =4 mm. In addition, inferior results were achieved at mandibular incisor and maxillary molar sites. Factors having no effect on root coverage included maxillary versus mandibular sites and single versus multiple adjacent sites. CONCLUSIONS: The use of guided tissue regeneration in periodontal plastic surgery is highly predictable, and highly esthetic root coverage can be gained without requiring a second surgical procedure or a second surgical site and is, therefore, an attractive alternative to conventional grafting techniques. PMID- 11063400 TI - Alveolar ridge and sinus augmentation utilizing platelet-rich plasma in combination with freeze-dried bone allograft: case series. AB - BACKGROUND: Alveolar bone regeneration is frequently necessary prior to placement of implants. Efforts to improve wound healing have focused on factors that may enhance bone formation following guided bone regeneration (GBR) techniques alone or in combination with bone replacement graft materials. Recent reports suggest that platelet-rich plasma (PRP), presumably high in levels of peptide growth factors, may enhance the formation of new bone when used in combination with autogenous graft material. METHODS: In this report, the clinical and radiographic results are presented on 15 consecutively treated patients using autologous PRP in combination with freeze-dried bone allograft (FDBA) for sinus elevation and/or ridge augmentation. FDBA and PRP (0.5 g/2cc PRP) were mixed and placed as a composite graft material. A gel formed by mixing autologous thrombin-rich plasma with PRP (1:4 ratio) was used to cover the graft material. Core biopsies of grafted areas were obtained in several patients as part of implant site preparation and were evaluated histologically to determine site maturation. RESULTS: Of 36 implant fixtures, 32 (89%) were considered clinically successful demonstrating complete bone coverage of the implant, no mobility, and a normal radiographic appearance at the time of re-entry and 12 months post-implant exposure. Four implants were removed due to mobility at the time of surgical exposure. Histologic evaluation of biopsy specimens revealed numerous areas of osteoid and bone formation around FDBA particles, with no evidence of inflammatory cell infiltrate. CONCLUSIONS: These clinical and histological findings suggest that ridge augmentation and sinus grafting with FDBA in combination with PRP provide a viable therapeutic alternative for implant placements. Future studies are necessary to determine whether PRP enhances new bone formation or maturation with bone replacement allografts. PMID- 11063401 TI - Are chlorinated pesticides a causation in maternal mitochondrial DNA (mtDNA) mutations? PMID- 11063402 TI - A paradigm for environmental epidemiology: why are effects of environmental exposures different from occupational effects? PMID- 11063403 TI - Pulmonary symptoms and spirometric values in Kangan Sour Gas Refinery workers. AB - In this study, the author measured the frequency of symptoms and/or alterations in respiratory functions in workers of the sour gas refining industry. All workers (n = 62) were employed in the most-exposed units of the Kangan Sour Gas Refinery. The refinery is approximately 250 km east of Bushehr Port along the Persian Gulf. This cross-sectional study involved a comprehensive health questionnaire, standardized clinical examinations by physicians blinded to subjects' symptoms and concerns, and multiple spirometric values. Although gas refinery workers experienced more respiratory symptoms than the 30 controls (i.e., 37.7% vs. 23.3%, respectively), who were matched for age and smoking status, pulmonary function data were not statistically different (p > .05) between the groups. The authors concluded, therefore, that in Kangan Sour Gas Refinery workers there were no respiratory or spirometric values associated with chronic low-dose exposure to sour gas plant emissions, including hydrogen sulfide. PMID- 11063404 TI - Environmental risk factors for respiratory infections. AB - In this study, the authors investigated the possible relationship between environmental-including dietary-factors and respiratory and ear infections in children. A sample of 304 children, aged 4-5 y, in Geneva, Switzerland, was studied. Mothers filled out a questionnaire that the authors provided. The questionnaire included items on food frequency, infections, and background factors (e.g., education, housing conditions, parents' smoking habits). Allergy was a risk factor for all infections, and mother's age was a protective factor. Humid conditions at home was a significant risk factor for cold, sore throat, and otitis (odds ratios = 2.71, 3.03, and 2.77, respectively); mold in the home was a significant risk factor for otitis (odds ratio = 2.80); and attending day-care centers was a significant risk factor for cold and bronchitis (odds ratios = 1.36 and 1.89, respectively). Dietary factors were not related to disease risk nor were environmental tobacco smoke or housing conditions generally related to an increased risk. PMID- 11063405 TI - Immunological and in-vivo neurological studies on a benzoic acid-specific T cell derived antigen-binding molecule from the serum of a toluene-sensitive patient. AB - T-cell-derived antigen-binding molecules (TABMs) specific for benzoic acid were isolated from the serum of a toluene-sensitive patient. The resulting purified TABMs (BA-TABMs) did not contain immunoglobulin G and were associated with the cytokine transforming growth factor-beta (TGF-beta). BA-TABMs bound to benzoic acid conjugated to human serum albumin (BA-HSA), as well as to other chemicals conjugated to human serum albumin-including dinitrophenol and oxazolone. The binding of BA-TABMs to the conjugated chemicals increased the level of detectable TGF-beta, and a similar effect was observed with the unconjugated chemicals, benzoic acid and 2,4-dinitrophenol glycine. The increase in TGF-beta was critically dependent on the ratio between BA-TABMs and the conjugated or unconjugated chemicals; the increase was optimum at intermediate concentrations and absent at low and high concentrations. The authors used an established animal model in vivo and demonstrated that TGF-beta enhanced the inflammatory response induced by the release of neuropeptides from sensory nerves; this enhancement occurred in a dose-dependent manner. The BA-TABMs also enhanced this neurogenic inflammatory response in a dose-dependent manner, and this effect was blocked by anti-TGF-beta antibody. When the authors added either BA-HSA or benzoic acid, the effect of BA-TABMs on neurogenic inflammation was further enhanced at intermediate concentrations of antigen and was unaltered or reduced at higher concentrations. TABMs specific to particular chemicals, as a result of their association with cytokines (e.g., TGF-beta), may be implicated in symptom production in chemically sensitive patients. PMID- 11063406 TI - Hypertension and chronic exposure to noise. AB - The effects of noise on various cardiovascular parameters are conflicting and uncertain. In the current study, the authors studied 52 workers who were employed in a bedframe factory who were chronically exposed to noise and who had poor hearing. An additional group of 65 workers who had jobs in the light-metal sector and another group of 64 office workers served as two control groups; none of the controls were exposed to noise, and none had hearing defects. Blood pressure was measured for each person in the supine and standing positions, and an electrocardiogram was also performed. Sound-level measurements were taken in the workplaces. Mean systolic and diastolic blood pressures and diastolic blood pressure distributions were significantly higher in the noise-exposed group than in both control groups. Among the three groups, there were significantly different frequencies of hypertension, drops in blood pressure, and electrocardiogram anomalies. Within the group of bedframe workers, those exposed to a personal daily level of exposure (i.e., equivalent continuous noise level for exposure to noise for each individual workers in an 8-hr shift) that exceeded 90 dBA had a higher mean diastolic blood pressure and a higher frequency of diastolic hypertension than workers exposed to a personal daily level of exposure of < 90 dBA. The findings suggested that (a) work performed by the bedframe group had some effects on the cardiovascular system, (b) noise is a cardiovascular risk factor, and (c) cardiovascular effects are relative to intensity and type of exposure. Vascular damage often accompanies auditory damage, but--depending on individual susceptibility--the cardiovascular system can respond in various ways. PMID- 11063407 TI - A case-control study of nitrate in drinking water and non-Hodgkin's lymphoma in Minnesota. AB - Nitrate in drinking water has been implicated as a possible risk factor for non Hodgkin's lymphoma. The authors examined the association between non-Hodgkin's lymphoma and waterborne nitrate through a population-based case-control study of white men in Minnesota. The authors, by linking residential histories with community water records, estimated average long-term exposure to nitrate in drinking water from 1947 to 1975 for 73 cases diagnosed between 1980 and 1982 and for 147 controls who used community water supplies. No association was found between nitrate levels in community water supplies and non-Hodgkin's lymphoma within the range of study exposures (median of highest exposure category = 2.4 mg nitrate/l [range = 0.1-7.2 mg/l]). The findings provide some safety assurance for those who use water systems that have nitrate levels that are less than 2.4 mg/l. PMID- 11063408 TI - Interactions between environmental lead exposure and sociodemographic factors on cognitive development. AB - A total of 375 children who lived in Port Pirie, South Australia, and surrounding towns were followed from birth to ages 11-13 y. Possible interactions between lifetime average blood lead concentration and sociodemographic factors (including gender, parents' occupational prestige [as a surrogate of socioeconomic status], quality of home environment, and maternal intelligence quotient) on children's intelligence quotients were examined. Although no statistically significant interaction between blood lead concentration and any of these covariates was found, the results suggested that-after adjustment for a wide range of covariates -children from socially disadvantaged backgrounds (adjusted regression coefficient = -9.6 intelligence quotient points per log unit of blood lead concentration; 95% confidence interval = -2.5, -17.7) were more sensitive to the effects of lead than those of a higher socioeconomic status (adjusted regression coefficient = -2.9; 95% confidence interval = 3.8, -9.6). In addition, girls (adjusted regression coefficient = -7.4; 95% confidence interval = -1.7, -13.1) were more sensitive to the effects of lead than boys (adjusted regression coefficient = -2.6; 95% confidence interval = 2.9, -8.0). These results were basically consistent with our findings observed at ages 2 y, 4 y, and 7 y. PMID- 11063409 TI - Health-care workers and latex allergy. AB - Latex hypersensitivity can pose a threat to anyone, but health-care providers are among the high-risk groups for developing latex hypersensitivity. Latex hypersensitivity likely results from health-care workers' increased use of gloves following implementation of universal precautions. It is also believed that the antigenic load of latex gloves causes an increase in latex hypersensitivity resulting from massive production of gloves. Although there are many studies on the prevalence of latex hypersensitivity among health-care workers, there appear to be discrepancies, which may affect the different apparent prevalence. Testing for latex hypersensitivity raises another problem. Latex allergens cannot be identified specifically; therefore, there is no standard test or testing solution that can identify hypersensitive persons. Although latex glove hypersensitivity was first identified in the late 1970s, there remain many uncertainties associated with it; as a result, there is a growing concern among health-care providers. The authors offer several precautions to avoid the development of latex hypersensitivity. PMID- 11063410 TI - Monitoring of low level arsenic exposure during maintenance of ion implanters. AB - To delineate potential exposure in ion implanter maintenance, the authors recruited 21 maintenance engineers (exposed group) and 10 computer programmers (controls) at three semiconductor manufacturing facilities. Samples of air, wipes, and urine; used cleaning cloths; and used gloves were collected for the characterization of arsenic exposure. Arsenic levels were very low in environmental samples, but high arsenic levels were found in some wipe samples, used cleaning cloths, and gloves. The average baseline content of urinary arsenic measured for maintenance engineers was 3.6 microg/g creatinine. Maintenance engineers experienced an increase of 1.0-7.8 microg/g creatinine in urinary arsenic levels during ion implanter maintenance. Results of a mixed-model analysis indicated that urinary arsenic levels were associated significantly with time series (p = .0001), and the extent of association was different among the three facilities (p = .0226). The results of this study indicate that arsenic intake via ingestion, rather than through inhalation, might play a significant role in the elevation of urinary arsenic levels. However, a series of urine samples with self-reference continue to be a good approach for the monitoring of low-level arsenic exposure. PMID- 11063411 TI - Determinants of blood lead levels across the menopausal transition. AB - In this study, the authors sought to evaluate the impact of menopause on lead remobilization from bone-lead stores. The study was conducted between 1993 and 1995 in Mexico City and included 903 women (mean age = 46.8 y [standard deviation = 8.2 y]). Participants provided information about reproductive variables and known risk factors for high PbB levels. PbB levels were determined with graphite furnace atomic absorption spectrophotometry. The authors used linear-regression models to describe the relationship between PbB levels and variables of interest. PbB levels ranged from 1.0 microg/dl to 43.8 microg/dl (mean = 11.0 microg/dl). Menopausal women at baseline had the highest PbB levels; the mean difference between pre- and postmenopausal women was 0.76 microg/dl (95% confidence interval = 0.024, 1.48). We observed an inverted U-shaped relationship between PbB level and age. The highest PbB levels were observed in women aged 47-50 y. Other important predictors of PbB levels were use of lead-glazed ceramics, number of pregnancies, history of cigarette smoking, and height. Our results support the hypothesis that bone lead may be mobilized during menopause and may constitute an important source of exposure. PMID- 11063412 TI - Identities and locations of 12 uranium mills in 1959 Atomic Energy Commission report. PMID- 11063413 TI - Arsenic residues in well water 36 y after endemic arsenic poisoning. PMID- 11063414 TI - Toxic metals in hens' eggs in India: a preliminary report. PMID- 11063415 TI - Interpenetrating polymer networks of alginate and polyethylene glycol for encapsulation of islets of Langerhans. AB - A mixture of alginate and polyethylene glycol acrylate was investigated as a system for the encapsulation of islets of Langerhans. This system showed dual crosslinkability: the alginate was ionically crosslinked by multivalent cations, and the PEG was covalently crosslinked by photoactivated free radical polymerization. The major advantage of the dually crosslinkable system was the chemical stability of the resultant gels due to the presence of covalent bonds that maintain the integrity of the gel as opposed to reversible ionic linkages that were the only mode of crosslinkage in previous generations of alginate-based encapsulation systems. The physical aspects of gelation of such alginate/PEG compositions were investigated. Diffusion of dextrans of known molecular weights through these gels was studied in order to shed light on the hydrogel porosity and permeability. In vitro viability and function tests demonstrated that these gels were biocompatible. Islets encapsulated in these systems were healthy and retained both viability and insulin secretory function. PMID- 11063416 TI - Control of molecular weight cut-off for immunoisolation by multilayering glycol chitosan-alginate polyion complex on alginate-based microcapsules. AB - Glycol chitosan is a positively charged polysaccharide which is water-soluble at pH 7.4, and is able to form a polyion complex (PIC) with anionic polymers, such as alginate. The authors attempt to develop a novel type of alginate-based microcapsule using this glycol chitosan for a islets-encapsulated bioartificial pancreas. The number of layers composed of glycol chitosan-alginate (GC-Alg) PIC were optimized, in order to cut off immunoglobulin transport and to protect encapsulated islets from the host immune reaction, and the transport characteristics were evaluated of glucose, bovine serum albumin (BSA) and gamma globulin. To add mechanical stability to the microcapsule, calcium ions, which crosslinked the alginate polymers close to the interface between core Ca-alginate and multilayered membrane, were partially substituted with barium ions after the formation of multilayered Ca-alginate gel beads. The partition coefficients of BSA and gamma-globulin were decreased with the increasing number of layers. The immunoisolation was achieved against gamma-globulin with four layers of the GC Alg PIC membrane, while BSA could permeate the membrane. The four-layered Ba alginate gel bead had a good permeability for glucose, giving a diffusion coefficient corresponding to 80% of that in pure water. Insulin secretion from the islets in the four-layered Ba-alginate microcapsule was satisfactorily observed with the fractional stimulation ratio of 2.17. This result indicates that the encapsulated islets maintained their viability even after encapsulation. It was, thus, shown that the Ba-alginate microcapsule with four layers of the GC Alg PIC membrane is promising as the microencapsulation material for a bioartificial pancreas. PMID- 11063417 TI - Can kinetic analysis be a tool for evaluating pore characteristics? AB - As the pore morphology influences drug release, the purpose is to study pore characteristics by comparing bead performances. Casein/gelatin beads have been prepared by the emulsification extraction method, cross-linked with D,L glyceraldehyde in acetone:water mixture 3:1 (v/v) and loaded with sodium fluorescein as a model drug. The beads with higher casein percentage have a higher matrix porosity, a wider average pore diameter and a higher cross-linking degree. The higher casein percentage causes a lower drug release rate. The kinetic analysis shows that the drug release occurs by diffusion and that the diffusion coefficient is affected by the casein percentage and the cross-linking degree. It can be hypothesized that the pore and channel morphology (tortuosity), due to the casein percentage in the matrix and the cross-linking treatment, can be evaluated by kinetic analysis of the release data. PMID- 11063418 TI - Tabletted polylactide microspheres prepared by a w/o emulsion-spray drying method. AB - An emulsification-spray drying technique is used to prepare poly(D,L-lactic acid) (PDLLA) microparticles loaded with a water soluble, non-steroidal anti inflammatory drug (NSAID), sodium naproxen (NaNPX). The method involves the preparation of a w/o emulsion in which the water soluble drug is dissolved in the aqueous dispersed phase, while the polymer is dissolved in the organic continuous phase. As a comparison, microparticles were prepared by spraying a suspension of the drug into an organic solution of the polymer. The spray-dried particles were characterized using SEM, DSC, XRD and in vitro release tests. The spray-dried product was then compressed (direct compression) to obtain controlled release matrix tablets. All microparticles release NaNPX within 30 min. The matrix tablets release the drug in 8-10 h; the matrix tablets characterized by the presence of surfactant (due to the emulsion used to obtain the microparticles) have the highest release rate. PMID- 11063419 TI - In vitro controlled release of bupivacaine from albumin microspheres and a co matrix formed by microspheres in a poly(lactide-co-glycolide) film. AB - Albumin microspheres cross-linked with glutaraldehyde and loaded with bupivacaine, a local anaesthetic, were synthesized (138 +/- 59 microm diameter). A matrix formed by bupivacaine-loaded microspheres in a poly(lactide-co glycolide) film was prepared in order to improve the controlled release of the drug. In vitro release of the drug was determined in phosphate buffer at 37 degrees C in the absence and in the presence of protease type VIII to mimic a biological system. The effect of temperature and protease on bupivacaine as a function of time was examined; both of them cause a degradative effect on the drug. A rapid release (60 +/- 8% of the drug) takes place at 1 h, and maximum release is found at 50 +/- 6 h from microspheres with swelling. In the presence of protease, maximum release of bupivacaine from microspheres is found at 28 +/- 2 h; the microspheres disappear at 8 days. Inclusion of bupivacaine-loaded microspheres in a poly(lactide-co-glycolide) film causes a slower release of the drug, up to 18 days, with swelling. In the presence of protease, the polymer protects bupivacaine-loaded microspheres from degradation, which takes place at 20 days. PMID- 11063420 TI - Drug particle size influence on enteric beads produced by a droplet extrusion/precipitation method. AB - The influence of drug particle size on the production of enteric beads by a polymer precipitation technique was investigated. Drug particle dimensions are known to play an important role in most microencapsulation techniques. Bead morphology was greatly influenced by drug particle size, and spherical shaped beads could only be obtained after size reduction of nimesulide crystals. This is confirmed by the angle of repose measurements, which show a significant decrease in theta values when beads are formulated with smaller drug particles. Furthermore, results show that drug encapsulation efficiency and in vitro drug release rates are also greatly dependent on both drug particle size and drug/polymer ratio in the initial suspension. Preparations containing 10.2 microm drug particles show a two-fold increase in the release rates when compared to those prepared with 40 microm particles. PMID- 11063421 TI - The stability of insulin in biodegradable microparticles based on blends of lactide polymers and polyethylene glycol. AB - Insulin-loaded microparticles were produced from blends of poly(ethylene glycol) (PEG) with poly (L-lactide) (PLA) homopolymer and poly (DL-lactide co-glycolide) copolymers (PLG) using a water-in-oil solvent extraction method. The dispersed phase was composed of PLG/PEG or PLA/PEG dissolved in dichloromethane, and the continuous phase was methanol containing 10% PVP. Characteristics, including particle size distribution, insulin loading capacity and efficiencies, in vitro release, degradation and stability, were investigated. The stability of insulin associated with microparticles prepared using PEG and 50:50 PLG and PLA was analysed by HPSEC and quantified by peak area following incubation in PBS at 37 degrees C for up to 1 month. Insulin was successfully entrapped in the PLG/PEG and PLA/PEG microparticles with trapping efficiencies up to 56 and 48%, loading levels 17.8 and 10.6% w/w, and particle sizes 8 and 3 microm, respectively. The insulin-loaded PLG/PEG and PLA/PEG microparticles were capable of controlling the release of insulin over 28 days with in vitro delivery rates of 0.94 and 0.65 microg insulin/mg particles/day in the first 4 days and a steady release with rate of 0.4 and 0.43 microg insulin/mg particles/day over the following 4 weeks, respectively. Extensive degradation of the PLG/PEG microparticles also occurred over 4 weeks, whereas the use of PLA/PEG blends resulted in a stable microparticle morphology and much reduced fragmentation and aggregation of the associated insulin. PMID- 11063422 TI - Microencapsulation of oils using sodium alginate. AB - The feasibility of encapsulating wheatgerm oil and evening primrose oil using sodium alginate by the emulsification method was explored in this study. It is based on the chemical reaction between the water-soluble sodium alginate and polyvalent cation, calcium, to form the water-insoluble alginate. The factors investigated were the physical appearance of the microspheres, the amount of oil that could be encapsulated, the flow property, size distribution and mean size of the microspheres produced. Encapsulation efficiency and oil content of wheatgerm oil increased with an increase in oil load. The mean size of the microspheres increased sharply at a high oil load of 250% w/w. Photographs of microspheres taken showed that the microspheres were larger, spherical and had more vesicles within, as oil load increased. Encapsulation efficiency of evening primrose oil microspheres was similar to wheatgerm oil microspheres at the respective oil loads of 50, 250 and 350% w/w. The emulsification method developed was successfully applied to wheatgerm oil, a fixed oil, with a maximum encapsulation efficiency of approximately 88%. It was satisfactory for evening primrose oil, which also belongs to the family of fixed oils. PMID- 11063423 TI - Preparation and characterization of microencapsulated gelospheres for controlled oral theophylline delivery. AB - Microencapsulated gelospheres were prepared using the water swellable polymers, poly(vinyl alcohol) and polyacrylamide in which the drug was embedded. Polymeric coating was formed by interfacial polymerization using 1,6 hexamine and sebacoyl chloride. The size, shape, in vitro release, kinetics and in vivo efficiency of the formulation were determined. The shape was found to be spherical with a size range below 100 microm. The per cent drug loaded was found to be higher in the case of gelospheres prepared with polyacrylamide than those prepared with poly(vinyl alcohol). The release rate was found to be near zero order. The AUC was found to be higher in the case of polyacrylamide and polyvinyl alcohol gelospheres as compared to plain drug solution. PMID- 11063424 TI - Preparation and in vitro characterization of gelatin microspheres containing Levodopa for nasal administration. AB - Transnasal absorption of pharmaceutical drugs has been recognized as an interesting alternative to the more conventional routes of administration. The aim of this paper was to develop a method of administrating L-dopa following the transnasal route. Gelatin microspheres were prepared by the w/o emulsification solvent extraction technique: the microspheres had a median particle size of 16.2 +/- 4.2 microm and were prepared using a stirring speed of 600 rpm for 5 min at 80 degrees C. The microspheres obtained were spherical and smooth-surfaced, and the microsphere size was inversely proportional to stirring speed (300-700 rpm) and to the percentage of the emulsifier (Tween 85, 1.4-2.7% v/v). L-dopa was incorporated into the microspheres with an efficiency of 65 +/- 6.7%. L-dopa was released from the microspheres, showing an initial fast release rate, followed by a second slower release rate. PMID- 11063426 TI - Literature alerts. PMID- 11063425 TI - Rifampicin polylactic acid microspheres for lung targeting. AB - Rifampicin polylactic acid microspheres for lung targeting were prepared by a modified emulsion-solvent diffusion method. The microspheres were free flowing, spherical with regular surface. Drug content, particle size distribution and in vitro release properties of the prepared microspheres were evaluated. In vivo experiments on rabbits showed remarkable accumulation of microspheres in the lung. PMID- 11063427 TI - Defining overweight and obesity: what are the issues? PMID- 11063428 TI - Addressing research questions with national survey data-the relation of vitamin A status to infection and inflammation. PMID- 11063429 TI - Human milk, fatty acids, and the immune response: a new glimpse. PMID- 11063430 TI - Effects of alcohol consumption on bone metabolism in elderly women. PMID- 11063431 TI - Criteria for definition of overweight in transition: background and recommendations for the United States. AB - Overweight and obesity are leading nutrition-related disorders of clinical and public health concern. Assessment and classification of these conditions are dependent on specific body mass index (BMI; in kg/m(2)) cutoff points. US government agencies are making the transition to a revised BMI definition of overweight from that previously recommended for general use. The purpose of this article is to inform the broader medical and scientific communities of the transition that is underway in the United States to identify and classify overweight among adults by using BMI. Historical background on the use of BMI in a variety of applications, as reported in US federal government agency documents, provides an understanding of previous and current weight-for-height guidelines and the basis for arriving at them. On the basis of the current Dietary Guidelines for Americans, US government agencies are moving toward the use of criteria for overweight and obesity that are consistent with current international standards. Clinicians, researchers, and journal editors should be aware of the transition toward a common definition of healthy weight, overweight, and obesity. To facilitate comparisons and reporting of data, others are encouraged to consider making this transition as well. PMID- 11063432 TI - Why and how should we measure oxidative DNA damage in nutritional studies? How far have we come? AB - Free radicals and other reactive species are constantly generated in vivo and cause oxidative damage to DNA at a rate that is probably a significant contributor to the age-related development of cancer. Agents that decrease oxidative DNA damage should thus decrease the risk of cancer development. That is, oxidative DNA damage is a "biomarker" for identifying persons at risk (for dietary or genetic reasons, or both) of developing cancer and for suggesting how the diets of these persons could be modified to decrease that risk. This biomarker concept presupposes that we can measure oxidative damage accurately in DNA from relevant tissues. Little information is available on whether oxidative DNA damage in blood cells mirrors such damage in tissues at risk of cancer development. Measurement of 8-hydroxylated guanine (eg, as 8-hydroxy-2' deoxyguanosine; 8OHdG) is the commonest method of assessing DNA damage, but there is no consensus on what the true levels are in human DNA. If the lowest levels reported are correct, 8OHdG may be only a minor product of oxidative DNA damage. Indeed, 8OHdG may be difficult to measure because of the ease with which it is formed artifactually during isolation, hydrolysis, and analysis of DNA. Mass spectrometry can accurately measure a wide spectrum of DNA base damage products, but the development of liquid chromatography-mass spectrometry techniques and improved DNA hydrolysis procedures is urgently required. The available evidence suggests that in Western populations, intake of certain fruit and vegetables can decrease oxidative DNA damage, whereas ascorbate, vitamin E, and beta-carotene cannot. PMID- 11063433 TI - Do adaptive changes in metabolic rate favor weight regain in weight-reduced individuals? An examination of the set-point theory. AB - BACKGROUND: Obese persons generally regain lost weight, suggesting that adaptive metabolic changes favor return to a preset weight. OBJECTIVE: Our objective was to determine whether adaptive changes in resting metabolic rate (RMR) and thyroid hormones occur in weight-reduced persons, predisposing them to long-term weight gain. DESIGN: Twenty-four overweight, postmenopausal women were studied at a clinical research center in four 10-d study phases: the overweight state (phase 1, energy balance; phase 2, 3350 kJ/d) and after reduction to a normal-weight state (phase 3, 3350 kJ/d; phase 4, energy balance). Weight-reduced women were matched with 24 never-overweight control subjects. After each study phase, assessments included RMR (by indirect calorimetry), body composition (by hydrostatic weighing), serum triiodothyronine (T(3)), and reverse T(3) (rT(3)). Body weight was measured 4 y later, without intervention. RESULTS: Body composition-adjusted RMR and T(3):rT(3) fell during acute (phase 2) and chronic (phase 3) energy restriction (P: < 0.01), but returned to baseline in the normal weight, energy-balanced state (phase 4; mean weight loss: 12.9 +/- 2.0 kg). RMR among weight-reduced women (4771 +/- 414 kJ/d) was not significantly different from that in control subjects (4955 +/- 414 kJ/d; P: = 0.14), and lower RMR did not predict greater 4-y weight regain (r = 0.27, NS). CONCLUSIONS: Energy restriction produces a transient hypothyroid-hypometabolic state that normalizes on return to energy-balanced conditions. Failure to establish energy balance after weight loss gives the misleading impression that weight-reduced persons are energy conservative and predisposed to weight regain. Our findings do not provide evidence in support of adaptive metabolic changes as an explanation for the tendency of weight-reduced persons to regain weight. PMID- 11063434 TI - HDL-cholesterol-raising effect of orange juice in subjects with hypercholesterolemia. AB - BACKGROUND: Orange juice-a rich source of vitamin C, folate, and flavonoids such as hesperidin-induces hypocholesterolemic responses in animals. OBJECTIVE: We determined whether orange juice beneficially altered blood lipids in subjects with moderate hypercholesterolemia. DESIGN: The sample consisted of 16 healthy men and 9 healthy women with elevated plasma total and LDL-cholesterol and normal plasma triacylglycerol concentrations. Participants incorporated 1, 2, or 3 cups (250 mL each) of orange juice sequentially into their diets, each dose over a period of 4 wk. This was followed by a 5-wk washout period. Plasma lipid, folate, homocyst(e)ine, and vitamin C (a compliance marker) concentrations were measured at baseline, after each treatment, and after the washout period. RESULTS: Consumption of 750 mL but not of 250 or 500 mL orange juice daily increased HDL cholesterol concentrations by 21% (P: < 0.001), triacylglycerol concentrations by 30% (from 1.56 +/- 0.72 to 2.03 +/- 0.91 mmol/L; P: < 0.02), and folate concentrations by 18% (P: < 0.01); decreased the LDL-HDL cholesterol ratio by 16% (P: < 0.005); and did not affect homocyst(e)ine concentrations. Plasma vitamin C concentrations increased significantly during each dietary period (2.1, 3.1, and 3.8 times, respectively). CONCLUSIONS: Orange juice (750 mL/d) improved blood lipid profiles in hypercholesterolemic subjects, confirming recommendations to consume >/=5-10 servings of fruit and vegetables daily. PMID- 11063435 TI - Plasma lipid profiles in adults after prenatal exposure to the Dutch famine. AB - BACKGROUND: Small body size at birth has been reported to be associated with an atherogenic lipid profile in humans, and animal experiments have shown that undernutrition during pregnancy permanently alters cholesterol metabolism in the offspring. There is no direct evidence in humans that maternal malnutrition during pregnancy affects the lipid profiles of the offspring. OBJECTIVES: We assessed the effects of maternal malnutrition during specific periods of gestation on plasma lipid profiles in persons aged approximately 50 y. DESIGN: This was a follow-up study of men and women born at term as singletons in a university hospital in Amsterdam between 1 November 1943 and 28 February 1947 around the time of a severe famine. RESULTS: Persons exposed to famine in early gestation had a more atherogenic lipid profile than did those who were not exposed to famine in utero. Their LDL-HDL cholesterol ratios were significantly higher (by 13.9%; 95% CI: 2.6-26.4%). Additionally, their plasma HDL-cholesterol and apolipoprotein A concentrations tended to be lower, and their plasma total cholesterol, LDL-cholesterol, and apolipoprotein B concentrations tended to be higher, although these differences were not statistically significant. The effect of famine was independent of size at birth and adult obesity. CONCLUSIONS: An atherogenic lipid profile might be linked to a transition from poor maternal nutrition in early gestation to adequate nutrition later on. This suggests that maternal malnutrition during early gestation may program lipid metabolism without affecting size at birth. PMID- 11063436 TI - Heavy coffee consumption and plasma homocysteine: a randomized controlled trial in healthy volunteers. AB - BACKGROUND: An elevated plasma concentration of total homocysteine is considered to be a strong risk factor for cardiovascular disease. Heavy coffee drinking has been related to high homocysteine concentrations in epidemiologic studies and in one experiment in which healthy subjects drank unfiltered, boiled coffee. OBJECTIVE: Our goal was to determine whether daily consumption of paper-filtered coffee raises plasma concentrations of total homocysteine in healthy subjects. DESIGN: Twenty-six volunteers (18-53 y of age) consumed 1 L/d of paper-filtered coffee brewed with 70 g regular ground beans or no coffee for 4 wk each in a randomized, crossover design. RESULTS: The mean (+/-SD) plasma concentration of total homocysteine in fasting blood was 8.1 +/- 1.8 micromol/L after abstention from coffee and 9.6 +/- 2.9 micromol/L after 3-4 wk of coffee drinking, a difference of 1.5 micromol/L (95% CI: 0.9, 2.1 micromol/L) or 18% (P: < 0.001). Coffee increased homocysteine concentrations in 24 of 26 individuals. Circulating concentrations of vitamin B-6, vitamin B-12, and folate were unaffected. CONCLUSION: Drinking large quantities of paper-filtered coffee raises fasting plasma concentrations of total homocysteine in healthy individuals. PMID- 11063437 TI - Diet-induced change in fatty acid composition of plasma triacylglycerols is not associated with change in glucagon-like peptide 1 or insulin sensitivity in people with type 2 diabetes. AB - BACKGROUND: Polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs) have been shown to positively affect blood lipids; however, their comparative effects on insulin sensitivity are unclear. OBJECTIVE: Our objective was to investigate whether chronic intake of MUFAs or PUFAs improves insulin sensitivity in people with type 2 diabetes via stimulation of the endogenous gut hormone glucagon-like peptide 1 [7-36] amide (GLP-1). DESIGN: Nine overweight people with type 2 diabetes received isoenergetic high-MUFA (20.3 +/- 3.5% of total energy) or high-PUFA (13.4 +/- 1. 3%) diets for 24 d in a randomized, double-blind crossover design. RESULTS: Weight and glycemic control remained stable throughout the study. Despite a significant change in the plasma triacylglycerol linoleic-oleic acid ratio (L:O) with both diets (MUFA: from 0.46 +/- 0.03 to 0.29 +/- 0.02, P: < 0.005; PUFA: from 0.36 +/- 0.04 to 0.56 +/- 0.05, P: < 0.05) and the phospholipid L:O (1.7 +/- 0.1 to 2.0 +/- 0.3; P: = 0.04) during consumption of the PUFA diet, this change was not associated with a change in insulin sensitivity, measured by the short-insulin-tolerance test. There was a significant reduction in the ratio of total to HDL cholesterol during consumption of the PUFA diet (5.2 +/- 0.4 compared with 4.7 +/- 0.3; P: = 0.005) but no change with the MUFA diet. There was no change in the fasting or postprandial incremental area under the curve in response to an identical standard test meal for glucose, insulin, triacylglycerol, nonesterified fatty acids, or GLP-1. CONCLUSIONS: Over the 3-wk intervention period, diet-induced change in the triacylglycerol or phospholipid L:O was not associated with either increased stimulation of GLP-1 or a change in insulin sensitivity in people with type 2 diabetes. PMID- 11063438 TI - Predictors of postprandial triacylglycerol response in children: the Columbia University Biomarkers Study. AB - BACKGROUND: Predictors of postprandial lipemia have not been explored in children. OBJECTIVE: Our objective was to determine whether the postprandial triacylglycerol response is associated with low HDL-cholesterol and high fasting triacylglycerol concentrations and family history of early-onset ischemic heart disease (IHD) in children. DESIGN: We administered a standardized fat load (52.5 g fat/m(2)) to 60 children (mean age: 14.0 y), 20 with and 40 without a family history of early-onset IHD, and to 29 mothers, all recruited from families enrolled in the Columbia University Biomarkers Study. Plasma lipid and retinyl palmitate concentrations were measured in the fasting state and 3, 6, and 8 h after the oral fat load. RESULTS: In children, postprandial lipemia, as indicated by the incremental area under the triacylglycerol response curve, was associated with elevated fasting triacylglycerol concentrations (>/=1.13 mmol/L; P: < 0.01), with low fasting HDL-cholesterol concentrations (/=40 y). All subjects received 2 isoenergetic study diets assigned by using a randomized, balanced crossover design. One diet provided 17% of energy as fructose. The other diet was sweetened with glucose and was nearly devoid of fructose. Each diet was fed for 6 wk. Both diets were composed of common foods and contained nearly identical amounts of carbohydrate, protein, fat, fiber, cholesterol, and saturated, monounsaturated, and polyunsaturated fatty acids. All meals were prepared in the metabolic kitchen of the General Clinical Research Center. RESULTS: The responses to the study diets differed by sex. In men, the fructose diet produced significantly higher fasting, postprandial, and daylong plasma triacylglycerol concentrations than did the glucose diet. The daylong plasma triacylglycerol concentration after 6 wk of the fructose diet was 32% greater in men than the corresponding concentration during the glucose diet (P: < 0.001). The fructose diet had no significant effect on fasting or postprandial plasma triacylglycerol concentrations in women. The fructose diet also had no persistent effect on fasting plasma cholesterol, HDL cholesterol, or LDL cholesterol in either men or women. CONCLUSIONS: Dietary fructose was associated with increased fasting and postprandial plasma triacylglycerol concentrations in men. Diets high in added fructose may be undesirable, particularly for men. Glucose may be a suitable replacement sugar. PMID- 11063440 TI - Effect of fat-reduced diets on 24-h energy expenditure: comparisons between animal protein, vegetable protein, and carbohydrate. AB - BACKGROUND: Single-meal tests have shown that protein has greater thermogenic and satiating effects than does carbohydrate, which may be relevant for the prevention and treatment of obesity if these effects can be maintained over 24 h. OBJECTIVE: The effects of pork-meat protein, soy protein, and carbohydrate on 24 h energy expenditure were compared. DESIGN: Twelve young, healthy, overweight and mildly obese [body mass index (in kg/m(2)): 26-32] nonsmoking men participated in a randomized, single-blind, 3-way crossover study lasting 4 d. The intervention had a 1-10-wk washout period. The 3 isoenergetic intervention diets were as follows: pork diet (29% of energy as fat and 29% as protein, mainly from pork meat), soy diet (29% of energy as fat and 28% as protein, mainly from soy), and carbohydrate diet (28% of energy as fat and 11% as protein). Twenty-four-hour energy expenditure was measured in a respiratory chamber at baseline and on day 4 of each intervention period. RESULTS: Twenty-four-hour energy expenditure was higher with the pork than with the soy (248 kJ/d, 1.9%; P: = 0.05) or carbohydrate (492 kJ/d, 3.9%; P: < 0.0001) diet and higher with the soy than with the carbohydrate (244 kJ/d, 1.9%; P: < 0.05) diet. However, because of a higher satiating effect, energy intake was 10-15% lower during the chamber stay than at baseline (P: > 0.05) with all 3 diets. The differences in energy expenditure remained unchanged after adjustment for differences in 24-h energy balance. CONCLUSIONS: Substitution of carbohydrate with 17-18% of energy as either pork meat or soy protein produced a 3% higher 24-h energy expenditure. The animal protein in pork meat produced a 2% higher 24-h energy expenditure than did the vegetable protein in soy. PMID- 11063441 TI - Effects of long-term supplementation with moderate pharmacologic doses of vitamin E are saturable and reversible in patients with type 1 diabetes. AB - BACKGROUND: Vitamin E supplementation has been proposed as adjunctive therapy to counteract the increased LDL oxidation in diabetes and thus prevent or delay cardiovascular complications. OBJECTIVE: The objective of this study was to investigate the effect of a moderate pharmacologic dose of vitamin E for 0.05) differences in retinol, ACT, or AGP by sex or age. Some 797 children (32%) had retinol concentrations <0.7 micromol/L and 87 (4%) had retinol concentrations <0.35 micromol/L; 274 children (11%) had elevated ACT (>0.6 g/L) and 1141 (45%) had elevated AGP (>1.2 g/L). Retinol concentration correlated with ACT (r = -0.141), AGP (r = -0.138), and ferritin (r = -0.09) (all P: < 0.001), but stepwise multiple regression indicated that these 3 variables made a minimal although quantifiable contribution to the variance of retinol (ACT, r(2) = 0.02; all 3 variables, r(2) = 0.03). CONCLUSIONS: The transient depression in plasma retinol produced by subclinical infection increased the number of at-risk children by 10% (76 of 797) and 56% (49 of 87) for plasma retinol concentrations <0.7 and <0.35 micromol/L, respectively. In addition, dietary inadequacy may be responsible for retinol concentrations being approximately 16% lower in Pakistani children than in children in the United Kingdom, where dietary vitamin A is adequate. PMID- 11063445 TI - Serum retinol, the acute phase response, and the apparent misclassification of vitamin A status in the third National Health and Nutrition Examination Survey. AB - BACKGROUND: Serum retinol decreases transiently during the acute phase response and can thus result in misclassification of vitamin A status. OBJECTIVE: Our objective was to determine the prevalence of acute phase response activation in a representative sample of the US population, identify the factors associated with this activation, and determine whether persons with an active acute phase response have lower serum retinol concentrations. DESIGN: Data from the third National Health and Nutrition Examination Survey (NHANES III) were analyzed. A serum C-reactive protein (CRP) concentration >/=10 mg/L indicated an active acute phase response. RESULTS: Mean serum retinol was lowest in subjects aged <10 y and increased with age. Concentrations were higher in males than in females aged 20 59 y. The prevalence of a CRP concentration >/=10 mg/L was lowest in subjects aged <20 y (/=10 mg/L also increases with age, is 2-fold greater in females than in males aged 20-69 y, and is associated with common inflammatory conditions. Thus, inflammation appeared to contribute to the misclassification of vitamin A status in the NHANES III population, and serum CRP is useful in identifying subjects who may be misclassified. PMID- 11063446 TI - Improved iodine status is associated with improved mental performance of schoolchildren in Benin. AB - BACKGROUND: An adequate iodine supply in utero and shortly after birth is known to be crucial to an individual's physical and mental development. The question of whether iodine supplementation later in life can exert a favorable influence on the mental performance of iodine-deficient populations was addressed in various studies, but with contradictory results. OBJECTIVE: The aim of this study was to examine the effect of an improvement in iodine status on mental and psychomotor performance of schoolchildren (7-11 y) who were moderately to severely iodine deficient. DESIGN: The study, which was originally planned as a double-blind, randomized, placebo-controlled intervention, was carried out in an iodine deficient population of schoolchildren (n = 196) in northern Benin. As the population began to have access to iodized salt during the 1-y intervention period, the study population was split post hoc-on the basis of urinary iodine concentrations-into a group with improved iodine status and a group with unchanged iodine status. Changes in mental and psychomotor performance over the intervention period were compared. RESULTS: Children with increased urinary iodine concentrations had a significantly greater increase in performance on the combination of mental tests than did the group with no change in urinary iodine concentrations. CONCLUSIONS: An improvement in iodine status, rather than iodine status itself, determined mental performance in this population, which was initially iodine deficient. These findings suggest a "catch-up" effect in terms of mental performance. PMID- 11063447 TI - Prenatal compared with parental origins of adolescent fatness. AB - BACKGROUND: Differences in prenatal growth influence postnatal body fat. OBJECTIVE: The objective was to investigate the role of parental body composition on the "tracking" of adolescent fatness. DESIGN: The study population consisted of 1993 white subjects and their parents. Measurements were taken at birth and again at the age of either 15, 16, or 17 y. The newborns were classified in 3 groups: small for gestational age, appropriate for gestational age, and large for gestational age. The mothers and fathers of the adolescents were classified into low and high subgroups on the basis of measurements of body mass index (BMI). Similarly, the mothers of the adolescents were classified into lean and fat subgroups on the basis of measurements of triceps skinfold thickness. RESULTS: Heavy newborns became heavier or fatter adolescents only when the mother or father was also fat and, among heavy newborns, the risk of becoming fat adolescents was approximately 5.7 times higher when the mother was fat rather than lean. CONCLUSIONS: Large newborns become fat adolescents only when the mother or father is also overweight or fat (ie, has either a high BMI or large skinfold thickness). These findings suggest that fatness during adolescence is related to parental fatness but not to prenatal fatness. Therefore, preventing higher levels of adiposity among newborns is unlikely to reduce overall adiposity in adolescence. PMID- 11063448 TI - Usefulness of serum transferrin receptor and serum ferritin in diagnosis of iron deficiency in infancy. AB - BACKGROUND: The serum transferrin receptor (TfR) and the ratio of TfR to serum ferritin (TfR:SF) have been shown to be useful as early indicators of iron deficiency. OBJECTIVE: The objective of this study was to evaluate the performance of TfR and TfR:SF in the assessment of iron deficiency in infants and to analyze age-related changes in both variables. DESIGN: A total of 716 blood samples obtained from 515 healthy infants aged 8-15 mo were studied. RESULTS: In 144 samples in which all other laboratory indicators of iron status were within the reference range, the median and 95% CI for TfR and TfR:SF were 8.5 mg/L (95% CI: 5.9, 13.5) and 497 (95% CI: 134, 975), respectively. TfR and TfR:SF were significantly correlated with the other laboratory indicators of iron status. Furthermore, as the severity of iron deficiency progressed, there was a gradual increase in mean TfR concentration (P: < 0.00001; analysis of variance). The sensitivity of TfR > 13.5 mg/L and TfR:SF > 975 in the diagnosis of iron deficiency was 23.6% and 68.4%, respectively. The specificity was 98.3% and 63.3% for TfR and TfR:SF, respectively. The sensitivity and specificity of SF < 10 microg/L were 63.7% and 60.8%, respectively. Receiver operator characteristic analysis showed that TfR and TfR:SF were more accurate than was SF alone in the diagnosis of iron deficiency. CONCLUSIONS: TfR and TfR:SF showed age-related changes; TfR and TfR:SF appear to be better diagnostic tests for iron deficiency in infants than SF. PMID- 11063449 TI - Unbound vascular endothelial growth factor and its receptors in breast, human milk, and newborn intestine. AB - BACKGROUND: Human milk, rich in cytokines, may contain the potent permeability- and angiogenesis-promoting agent vascular endothelial growth factor (VEGF). OBJECTIVE: We wanted to study whether free or bound VEGF is present in human milk and whether it and its receptors (VEGFR-1 and -2) are expressed in lactating breast or newborn intestinal tissue. DESIGN: The study had a longitudinal design with collection of human milk from healthy (n = 32) and diabetic (n = 5) women at 2, 7, and 30 d postpartum. Milk was analyzed for VEGF by enzyme-linked immunosorbent assay along with plasma samples collected 2 d postpartum. Immunohistochemistry was used to localize VEGF and its receptors in lactating breast and newborn intestine. Gel filtration with radiolabeled VEGF was performed to study whether human milk contains VEGF binding proteins. RESULTS: Human milk VEGF concentrations in healthy (76 +/- 19 microg/L, x +/- SD) and diabetic (75 +/ 25 microg/L) women did not differ at 2, 7 (23 +/- 7 and 27 +/- 8 microg/L, respectively), or 30 d (14 +/- 5 and 17 +/- 7 microg/L, respectively) postpartum. VEGF was undetectable in all but 3 plasma samples. Human milk was free of VEGF binding proteins. VEGFR-1 and -2 immunoreactivity was seen in the glandular epithelial cells of the newborn intestine and lactating breast, whereas VEGF was present only in breast glandular epithelium. CONCLUSIONS: The high concentrations of VEGF in human milk, especially colostrum, are not affected by maternal diabetes and may play a role in newborn nutrition. PMID- 11063450 TI - Breast-fed and formula-fed infants do not differ in immunocompetent cell cytokine production despite differences in cell membrane fatty acid composition. AB - BACKGROUND: Breast-fed and formula-fed infants differ in the amount and type of polyunsaturated fatty acids consumed. The fatty acid composition of cell membranes is related to dietary fatty acids and, in adults, changes in membrane fatty acid composition are accompanied by changes in monocyte cytokine production and hence a modification of the immunologic response. OBJECTIVE: Our objective was to determine whether production by immunocompetent cells of the proinflammatory cytokines interleukin 1 (IL-1) and tumor necrosis factor (TNF) differs between breast-fed and formula-fed infants. DESIGN: Twenty-six healthy infants (13 breast-fed and 13 fed modified cow-milk formula) aged 2-4 mo were studied. The fatty acid composition of red blood cell (RBC) membrane phospholipids was measured by gas-liquid chromatography and IL-1 and TNF release were measured in whole blood culture in bacterial-endotoxin-stimulated and unstimulated cells. RESULTS: The infants' ages, weights, hemoglobin concentrations, and white blood cell counts did not differ significantly between groups. The percentage of n-3 fatty acids of total RBC phospholipid fatty acids was significantly higher in breast-fed than in formula-fed infants (6.31 +/- 2.5% compared with 2.98 +/- 0.97%); docosahexaenoic acid (22:6n-3) concentrations were also markedly higher in breast-fed infants (5.1 +/- 1.2% compared with 2.2 +/- 0.9%, P: < 0.001), but eicosapentaenoic acid (20:5n-3) and docosapentaenoic acid (22:5n-3) concentrations did not differ significantly between groups. The percentage of n-6 fatty acids was not significantly different between groups. The percentage of oleic acid (18:1) was higher in formula-fed than in breast-fed infants (16.2 +/- 0.7% compared with 20.6 +/- 1.1%; P: < 0.001). IL-1 and TNF release in whole blood culture did not differ significantly between groups. CONCLUSION: The release of proinflammatory cytokines by immunocompetent cells does not differ significantly in breast-fed and formula-fed infants despite differences in cell membrane fatty acid composition. PMID- 11063451 TI - Alcohol intake and bone metabolism in elderly women. AB - BACKGROUND: Published reports on the effect of alcohol consumption on bone mineral density (BMD) are inconsistent. OBJECTIVE: The objective of this study was to examine the relation between alcohol intake and BMD, calcitropic hormones, calcium absorption, and other biochemical indexes of bone and mineral metabolism in elderly women. DESIGN: The results presented are derived from baseline observations of 489 elderly women (aged 65-77 y) recruited for an osteoporosis study. The nondrinking group comprised 297 women and the drinking group comprised 148 women. Furthermore, the effect of different alcohol intakes (28.6 to 57.2 to 142.9 g/wk) was studied. RESULTS: Women who consumed alcohol had significantly higher spine (10%), total body (4.5%), and midradius (6%) BMD than did nondrinkers. An alcohol intake >28.6 g/wk was associated with higher BMD; maximum effect was seen with an intake of >28.6 to 2 y is sufficient for adequate growth. Lower fat intakes may be associated with inadequate vitamin and mineral intakes and increased risk of poor growth. Diets higher in fat may lead to higher energy intakes and higher body fat, although available data for children are conflicting. Beyond infancy, children can meet their energy needs for maintenance, physical activity, and growth from a diet providing 30% of energy from fat. PMID- 11063467 TI - Fat and energy needs of children in developing countries. AB - The fat requirements of children can be judged according to 4 criteria: 1) the possible obligate needs of fat as a metabolic fuel, 2) the provision of a sufficiently energy-dense diet to meet energy needs, 3) the adequate supply of essential fatty acids, and 4) the supply of sufficient fat to allow adequate absorption of fat-soluble vitamins. In these respects the fat requirements of children in developing countries are probably similar to those of children in affluent nations except for the additional needs imposed by environmental stresses, particularly recurrent infections. In many developing countries, the low energy density of weaning foods appears to be a major contributor to growth faltering and ultimate malnutrition. Evidence from doubly labeled water studies suggests that these diets are adequate when children are healthy but fail to support rapid catch-up growth after diarrhea and other infections. The issues in determining and meeting the fat needs of children in developing countries are illustrated with use of detailed comparative dietary data from a rural community in The Gambia and from Cambridge, United Kingdom. The outstanding feature of the Gambian data is the great importance of breast milk as a source of fat and essential fatty acids up until the end of the second year of life. Weaning foods and adult foods contain low amounts of fat, which causes a sharp transition from adequate fat intakes to probable inadequate fat intakes when children are weaned from the breast. The effects of such low fat intakes, particularly in terms of immune function, require investigation. PMID- 11063468 TI - Rapid westernization of children's blood cholesterol in 3 countries: evidence for nutrient-gene interactions? AB - The aim of this study was to examine potential factors that modify blood cholesterol among children in countries in which dietary and lifestyle habits are becoming westernized. Population data on serum total and lipoprotein cholesterol, anthropometric indexes, and dietary intake were reviewed and compared for children aged 1-18 y from Japan, Spain, and the United States. The data show that total serum cholesterol in Japanese and Spanish children recently exceeded the 75th percentile for US children, primarily reflecting LDL cholesterol, although both LDL and HDL cholesterol contributed. Adiposity indexes do not explain the trends observed. Total and saturated fat intakes increased substantially in both Japan and Spain but in Japan are still lower than intakes in the United States. The Hegsted equation was used to relate differences in serum cholesterol to dietary fat intake. Changes in total serum cholesterol followed established dietary correlations among children in Spain, but not in Japan. Serum cholesterol in Japanese children was predicted to be 0.20-0.32 mmol/L lower than in US children; actual concentrations were considerably higher. These results suggest that a rapid westernization of children's blood cholesterol concentrations has occurred in Japan and Spain. Changes in fat intake predict changes in blood cholesterol in Spain, but not in Japan. Differences in genetic response to diet in certain populations, such as the Japanese, may explain higher blood cholesterol concentrations with lower fat intakes compared with the United States. PMID- 11063469 TI - Influence of genetic polymorphisms on responsiveness to dietary fat and cholesterol. AB - Genes influence quantitative variations in plasma lipoprotein concentrations. For example, intake of dietary saturated fat and cholesterol raises the average serum cholesterol concentration, leading to a higher risk of coronary artery disease in populations. However, not all individuals within the population are susceptible: genetic factors appear to render individuals either "dietary responsive" or "dietary nonresponsive." In this review, we focus on current knowledge about the influence of genetic polymorphisms in certain genes on the lipoprotein response to dietary fat and cholesterol. Our preliminary studies in the Dietary Intervention Study in Children suggest a significant dose-response relation between the decrease in LDL cholesterol from baseline to 36 mo of follow-up in both the intervention group (who consumed a low-fat, low-cholesterol diet) and the usual care group (who consumed a regular diet) and the presence of the APOA1*A allele at the M1 site and the + site at the M2 site of the gene encoding apolipoprotein (apo) A-I. The DNA polymorphisms on the genes encoding apo A-IV, apo B, apo C-III, apo E, lipoprotein lipase, cholesteryl ester transfer protein, lecithin:cholesterol acyltransferase (phosphatidylcholine-sterol O: acyltransferase), and LDL receptor were found by others to be associated with the plasma lipoprotein response to dietary intervention. Possible mechanisms involved in these effects are discussed and certain discrepancies in the literature about some genetic effects on responsiveness are analyzed. An improved understanding of the influence of specific genes on lipoprotein responsiveness to dietary fat and cholesterol may allow us to identify and counsel certain individuals to avoid high-fat diets so that they may reduce their risk of developing hyperlipidemia and coronary artery disease. PMID- 11063470 TI - Gene-diet interactions in obesity. AB - A considerable amount of research on the genetics of obesity has been reported in the past few years. Despite evidence that genetic factors play a significant role in the etiology of this nutritional disease and the increasing number of obesity genes identified, relatively little is known about the role of genes in the response of obesity phenotypes to alterations in energy balance or diet composition. This is especially true for dietary fat, which is known to be associated with obesity at the population level. The aim of this review was to summarize the evidence currently available about the role of gene-nutrient interactions in human obesity. Evidence from both genetic epidemiology and molecular epidemiology studies suggests that genetic factors are involved in determining the susceptibility to gaining or losing fat in response to diet or the risk of developing some of the comorbidities generally observed in obese individuals. Recent evidence suggests that quantitative trait loci identified from animal models of diet-induced obesity could influence body fat in humans. Despite the limited number of studies, the evidence on gene-diet interactions in obesity is convincing. More research is needed to identify the genes responsible for these interaction effects, and the use of animal models of diet-induced obesity represents a promising approach. Finally, data on children are needed to allow assessment of the tracking of nutrient intake between childhood and adulthood. In addition, gene-diet interactions in children need to be investigated to determine whether the genes involved are the same as those found in adults. PMID- 11063471 TI - Dietary fat and adult diseases and the implications for childhood nutrition: an epidemiologic approach. AB - Reducing dietary saturated fat by 7% of energy, a realistic target, would reduce serum cholesterol by 10% and mortality from ischemic heart disease by 25-30%. Randomized trials show that this mortality reduction is attained rapidly, usually by the third year after initial reduction of dietary saturated fat intake. Dietary change in adulthood may therefore reverse the adverse health effects of a high-fat diet in childhood. In the absence of such change, however, dietary fat in childhood may increase the risk of cardiovascular disease in adult life because of a longer duration of exposure to a high-fat diet. Assessing the effects of diet on cancer risk is more difficult. The intermediary markers of risk that are analogous to serum cholesterol are less satisfactory and there are negligible trial data. Cohort studies of diet and cancer, although subject to bias, do not favor a direct causal relation between dietary fat and cancer. But a reduction in risk is likely when dietary fat is reduced as part of a general change toward a healthier diet. The trend toward increased energy intake and body size in childhood and relatively low dietary fiber contribute to the decreasing age at menarche, which is associated with an increased risk of breast cancer. Low dietary fiber, low fruit and vegetable consumption, and high red meat consumption are associated with colon cancer and other cancers, and important causal effects of diet on cancer are likely. As with cardiovascular disease, this dietary trend that is commenced in childhood is likely to increase age-specific rates of colon cancer in adult life, but the risk may be reversed with later dietary change. PMID- 11063472 TI - Lipid and macrophage accumulations in arteries of children and the development of atherosclerosis. AB - About one-half of infants in the first 6 mo of life have small collections of macrophages and macrophages filled with lipid droplets (foam cells) in susceptible segments of the coronary arteries. In subsequent years, fewer children have foam cells but around puberty (12-15 y) foam cell accumulations mostly larger than those in infants occur in 69% of adolescents. Lesions that represent the previously missing link between foam cell accumulations and atheromas have now been identified in a subgroup of highly susceptible locations. Such "preatheroma" lesions contain small pools of lipid droplets and dead cell remnants (extracellular lipid) in addition to macrophage foam cells. Atheromas, which emerge in some adolescents and young adults in the same locations, have a lipid core in which increased extracellular lipid displaces structural smooth muscle cells and the normal extracellular matrix. As soon as lipid cores form, calcium granules appear in some smooth muscle cells and among the extracellular lipid of the core. The degree of calcification is variable and, in youth, generally small. In the age group of 16-19 y, 15% of persons have either preatheromas or atheromas in coronary arteries; foam cell accumulations only are present in an additional 53% of 16-19-y-olds. Because the lipid cores of atheromas may be an underlying cause of lesion rupture, hematomas, and thrombosis, and because their development begins soon after puberty, it would be prudent to attempt to lower the influx of excessive atherogenic lipoproteins into the arterial wall by that age. PMID- 11063473 TI - Origin of atherosclerosis in childhood and adolescence. AB - Atherosclerosis begins in childhood as deposits of cholesterol and its esters, referred to as fatty streaks, in the intima of large muscular arteries. In some persons and at certain arterial sites, more lipid accumulates and is covered by a fibromuscular cap to form a fibrous plaque. Further changes in fibrous plaques render them vulnerable to rupture, an event that precipitates occlusive thrombosis and clinically manifest disease (sudden cardiac death, myocardial infarction, stroke, or peripheral arterial disease). In adults, elevated non-HDL cholesterol concentrations, low HDL-cholesterol concentrations, hypertension, smoking, diabetes, and obesity are associated with advanced atherosclerotic lesions and increased risk of clinically manifest atherosclerotic disease. Control of these risk factors is the major strategy for preventing atherosclerotic disease. To determine whether these risk factors also are associated with early atherosclerosis in young persons, we examined arteries and tissue from approximately 3000 autopsied persons aged 15-34 y who died of accidental injury, homicide, or suicide. The extent of both fatty streaks and raised lesions (fibrous plaques and other advanced lesions) in the right coronary artery and in the abdominal aorta was associated positively with non-HDL cholesterol concentration, hypertension, impaired glucose tolerance, and obesity and associated negatively with HDL-cholesterol concentration. Atherosclerosis of the abdominal aorta also was associated positively with smoking. These observations indicate that long-range prevention of atherosclerosis and its sequelae by control of the risk factors for adult coronary artery disease should begin in adolescence and young adulthood. PMID- 11063474 TI - Special Turku Coronary Risk Factor Intervention Project for Babies (STRIP). AB - BACKGROUND: Introducing nutritional and lifestyle principles to children in late infancy may permanently improve their adherence to a low-saturated fat, low cholesterol diet, thus reducing of coronary risk factors, but worries about possible effects on growth and development have hampered such an approach. OBJECTIVE: The Special Turku Coronary Risk Factor Intervention Project for Babies (STRIP) aimed to decrease exposure to known environmental atherosclerosis risk factors in children 7-36 mo of age. DESIGN: Repeated, individualized counseling aimed at promoting a fat intake of 30% of energy and a 1:1:1 ratio of saturated to monounsaturated to polyunsaturated fat intake was provided (n = 540 intervention children; 284 boys). Nutrition was discussed superficially with the families of the control children (n = 522; 266 boys) and food intake was recorded at 3-6-mo intervals by use of 3-4-d food diaries. Serum lipids were measured at 6 12-mo intervals and growth was monitored regularly. RESULTS: Fat intake of the intervention (control) children provided 29.5% (29.4%) of energy at the age of 8 mo, 26.6% (28.5%) of energy at 13 mo, 30.5% (33.5%) of energy at 24 mo, and 31. 5% (33.5%) of energy at 36 mo. The intervention children consistently consumed less saturated fat than did the control children (P: <0.0001). Recommended intakes of other nutrients (except vitamin D and occasionally iron) were reached irrespective of the amount and type of dietary fat. Serum cholesterol, non-HDL cholesterol, and HDL-cholesterol concentrations were 3-6% lower in the intervention children than in the control children. The intervention had no effect on height, weight, or head circumference gain. Fat intake did not predict children's growth patterns. CONCLUSION: Repeated, individualized counseling in early childhood aimed at reducing consumption of saturated fat and cholesterol was effective and feasible and did not restrict growth in circumstances in which children were regularly monitored. PMID- 11063475 TI - Efficacy and safety of lowering dietary intake of total fat, saturated fat, and cholesterol in children with elevated LDL cholesterol: the Dietary Intervention Study in Children. AB - BACKGROUND: Few studies have shown the efficacy and safety of lower-fat diets in children. OBJECTIVE: Our objective was to assess the efficacy and safety of lowering dietary intake of total fat, saturated fat, and cholesterol to decrease LDL-cholesterol concentrations in children. DESIGN: A 6-center, randomized controlled clinical trial was carried out in 663 children aged 8-10 y with LDL cholesterol concentrations greater than the 80th and less than the 98th percentiles for age and sex. The children were randomly assigned to either an intervention group or a usual care group. Behavioral intervention promoted adherence to a diet providing 28% of energy from total fat, <8% from saturated fat, 92%) and highly specific (>94%). SeroHSV2 does not cross-react with other alphaherpesvirus antibodies. SeroHSV1 is highly sensitive (>94%) and specific (>91%) compared to four commercial available kits. SeroHSV IgM is highly specific (>92%) in comparison with other commercial HSV IgM tests. The sensitivity of SeroHSV IgM ranges between 50 and 70% compared to these tests. Further investigation of the discrepant results obtained by using in-house competition tests indicated that SeroHSV IgM is more sensitive. SeroHSV IgG was also found to be highly sensitive (>94%) and highly specific (>92%) compared to the other commercial HSV IgG tests. PMID- 11063497 TI - Close association between pulmonary disease manifestation in Mycoplasma pneumoniae infection and enhanced local production of interleukin-18 in the lung, independent of gamma interferon. AB - To investigate pathophysiologies of Mycoplasma pneumoniae infection from an immunological point of view, we measured the levels of interleukin-18 (IL-18) (originally designated gamma interferon [IFN-gamma]-inducing factor) in 19 serum samples from 10 patients with pneumonia without pleural effusion (ages 1 to 16 years), 3 serum and 13 pleural fluid samples from 11 patients with pleural effusions (ages 11 months to 15 years), and 18 serum and 27 cerebrospinal fluid samples from 24 patients with central nervous system complications (ages 1 to 15 years). IL-18 was measured by a commercially available enzyme-linked immunosorbent assay kit (MBL, Nagoya, Japan). In addition, the levels of tumor necrosis factor alpha, IFN-gamma, IL-6, IL-12, and KL-6 (a mucin-like glycoprotein expressed on type 2 pneumocytes) were measured in selected samples. The results concerning pleural effusions showed that elevated levels of IL-18 in pleural fluid, but not in serum, were solely associated with a sustained fibrotic change of the lung on chest roentgenography which might represent a pathological feature of intraluminal organization. All the pleural fluid samples with elevated levels of IL-18 were positive by PCR for M. pneumoniae DNA. There was no association between IL-18 and IFN-gamma levels in serum or in the pleural fluid. On the other hand, elevated levels of IL-18 in serum, but not in cerebrospinal fluid samples, were observed in the cases complicated by central nervous system involvement, including profound brain dysfunction with seizures. Our study demonstrated that M. pneumoniae can induce IL-18 and that the enhanced local production of IL-18 in the lung is closely associated with pulmonary disease manifestation. PMID- 11063498 TI - Combined cell culture enzyme-linked immunosorbent assay for quantification of poliovirus neutralization- relevant antibodies. AB - A combined cell culture enzyme-linked immunosorbent assay (CCC-ELISA) was developed for measuring the neutralizing antipoliovirus antibodies in human sera. The binding of different concentrations of each of the three poliovirus types to BGM cells in the presence and absence of a constant dilution from each test and reference serum was measured in the CCC-ELISA. The titers of the viruses neutralized by each serum were measured with the titration curves and used for interpretation of neutralizing titers to the three poliovirus types. Analysis of human sera revealed that the sensitivity and specificity of the CCC-ELISA and the microneutralization assay were comparable. The CCC-ELISA is nonsubjective, rapid, and highly reproducible. Furthermore, the CCC-ELISA could potentially be used as a seroepidemiologic tool for assessment of the humoral response to the cell culture infectious viruses. PMID- 11063499 TI - Measurement of cytokine secretion, intracellular protein expression, and mRNA in resting and stimulated peripheral blood mononuclear cells. AB - Quantitation of cytokine production is a valuable adjunct to standard immunologic assays in defining several pathologic processes. Nevertheless, there is little agreement about which tissues should be assayed, which type of assay should be performed, and which stimulation protocol should be used. As these types of assays enter the clinical arena, there is need for standardization. There is also a need to maximize the amount of information which may be derived from a single sample. We compared secreted interleukin 4 (IL-4), IL-2, IL-6, tumor necrosis factor alpha (TNF-alpha), and gamma interferon proteins as measured by enzyme linked immunosorbent assay with intracellular cytokine production (IL-2 and gamma interferon) as detected by flow cytometry and quantitative competitive PCR for IL 2, IL-4, TNF-alpha, and gamma interferon mRNA and cDNA. Results from unstimulated cells and cells stimulated with phorbol myristate acetate, phytohemagglutinin, and phorbol myristate acetate plus phytohemagglutin were compared. All three methodologies detected significant stimulation of cytokine production. The combination of phytohemagglutinin and phorbol myristate acetate was overall the most-potent stimulus. PMID- 11063500 TI - Clarithromycin attenuates mastectomy-induced acute inflammatory response. AB - Based on the observation that administration of clarithromycin led to an attenuation of the inflammatory response induced by surgical trauma in a guinea pig model, we investigated the potential beneficial effects of clarithromycin on the local and systemic inflammatory response in patients undergoing mastectomy in an open-label prospective study. During a 16-month period, 54 patients who underwent mastectomy were randomly divided into two groups. In one group, the patients received oral clarithromycin at a dose of 500 mg twice a day, from the day before to 3 days after mastectomy. There was no significant difference in the incidence of antibiotic prophylaxis-related toxicities or postoperative infections between the patients who received clarithromycin and those who did not. Clarithromycin treatment was significantly associated with an attenuation of febrile response, tachycardia, tachypnea, and an increase in monocyte counts (P, <0.0001, <0.01, <0.05, and <0.01, respectively). Clarithromycin also reduced the intensity and duration of postoperative pain (P, <0.05 and <0.005, respectively) and increased the range of motion of the involved shoulder (P < 0.05 for abduction and flexion). We conclude that clarithromycin effectively modulates the acute inflammatory response associated with mastectomy and produces a better clinical outcome. PMID- 11063501 TI - Structural and immunological characteristics of a 28-kilodalton cruzipain-like cysteine protease of Paragonimus westermani expressed in the definitive host stage. AB - A complete cDNA sequence encoding a 28-kDa cruzipain-like cysteine protease of adult Paragonimus westermani, termed Pw28CCP, was isolated from an adult cDNA library. The cDNA contained a single open reading frame of 975 bp encoding 325 amino acids, which exhibited the structural motif and domain organization characteristic of cysteine proteases of non-cathepsin Bs including a hydrophobic signal sequence, an ERFNIN motif, and essential cysteine residues as well as active sites in the mature catalytic region. Analysis of its phylogenetic position revealed that this novel enzyme belonged to the cruzipain-like cysteine proteases. The sequence of the first 13 amino acids predicted from the mature domain of Pw28CCP was in accord with that determined from the native 28-kDa enzyme purified from the adult worm. Expression of Pw28CCP was observed specifically in juvenile and adult worms, with a location in the intestinal epithelium, suggesting that this enzyme could be secreted and involved in nutrient uptake and immune modulation. The recombinant protein expressed in Escherichia coli was used to assess antigenicity by immunoblotting with sera from patients with active paragonimiasis and from those with other parasitic infections. The resulting sensitivity of 86.2% (56 of 65 samples) and specificity of 98% (147 of 150 samples) suggest its potential as an antigen for use in immunodiagnosis. PMID- 11063502 TI - Detection and quantification of antibodies to Newcastle disease virus in ostrich and rhea sera using a liquid phase blocking enzyme-linked immunosorbent assay. AB - A liquid phase blocking ELISA (LPB-ELISA) was adapted for the detection and quantification of antibodies to Newcastle disease virus. Sera from vaccinated and unvaccinated commercial flocks of ostriches (Struthio camelus) and rheas (Rhea americana) were tested. The purified and nonpurified virus used as the antigen and the capture and detector antibodies were prepared and standardized for this purpose. The hemagglutination-inhibition (HI) test was regarded as the reference method. The cutoff point for the LPB-ELISA was determined by a two-graph receiver operating characteristic analysis. The LPB-ELISA titers regressed significantly (P < 0.0001) on the HI titers with a high correlation coefficient (r = 0.875). The two tests showed good agreement (kappa = 0.82; P < 0.0001), relative sensitivity (90.91%) and specificity (91.18%), and accuracy (91.02%), suggesting that they are interchangeable. PMID- 11063503 TI - Antifreeze solution improves DNA recovery by preserving the integrity of pathogen infected blood and other tissues. AB - Preserving blood samples for shipping and later DNA extraction has been performed by cooling, freezing, drying, freeze-drying, and protease treatment, among other methods. Most methods to preserve field samples for further DNA extraction do not prevent cellular and DNA damage or are useful only in preserving them for short periods. This report introduces a novel method for blood and tissue that allows preservation in freezing temperatures for a prolonged period of time. The solution reported here (20% ethylene glycol-propylene glycol) preserves cells and tissues integrity, as judged by microscopic analysis, and improves DNA yield and quality. PMID- 11063504 TI - Jasplakinolide induces apoptosis in various transformed cell lines by a caspase-3 like protease-dependent pathway. AB - To clarify the mechanisms underlying the antiproliferative effects of jasplakinolide, a cyclic depsipeptide from marine sponges, we examined whether jasplakinolide induces apoptosis in a variety of transformed and nontransformed cells. Jasplakinolide inhibited proliferation of human Jurkat T cells, resulting in cell death. This was accompanied by chromatin condensation and DNA cleavage at the linker regions between the nucleosomes. When caspase-3-like activity in the cytosolic extracts of Jurkat T cells was examined with a fluorescent substrate, DEVD-MAC (N-acetyl-Asp-Glu-Val-Asp-4-methyl-coumaryl-7-amide), the activity in the cells treated with jasplakinolide was remarkably increased in a time dependent manner. Pretreatment of Jurkat T cells with the caspase inhibitor zVAD [benzyloxycarbonyl(Cbz)-Val-Ala-beta-Asp(OMe)-fluoromethylketone] or DEVD-CHO (N acetyl-Asp-Glu-Val-Asp-1-aldehyde) prevented the induction of apoptosis by jasplakinolide. Moreover, exposure of various murine transformed cell lines to jasplakinolide resulted in cell death, which was inhibited by zVAD. Although it has been well established that murine immature thymocytes are sensitive to apoptosis when exposed to various apoptotic stimuli, these cells as well as mature T lymphocytes were resistant to jasplakinolide-induced apoptosis. The results suggest that jasplakinolide induces apoptotic cell death through a caspase-3-like protease-dependent pathway. Another important outcome is that transformed cell lines were more susceptible to jasplakinolide-induced apoptosis than normal nontransformed cells. PMID- 11063505 TI - T-Cell receptor Vbeta repertoire CDR3 length diversity differs within CD45RA and CD45RO T-cell subsets in healthy and human immunodeficiency virus-infected children. AB - The T-cell receptor (TCR) CDR3 length heterogeneity is formed during recombination of individual Vbeta gene families. We hypothesized that CDR3 length diversity could be used to assess the fundamental differences within the TCR repertoire of CD45RA and CD45RO T-cell subpopulations. By using PCR-based spectratyping, nested primers for all 24 human Vbeta families were developed to amplify CDR3 lengths in immunomagnetically selected CD45RA and CD45RO subsets within both CD4(+) and CD8(+) T-cell populations. Umbilical cord blood mononuclear cells or peripheral blood mononuclear cells obtained from healthy newborns, infants, and children, as well as human immunodeficiency virus (HIV) infected children, were analyzed. All T-cell subsets from newborn and healthy children demonstrated a Gaussian distribution of CDR3 lengths in separated T-cell subsets. In contrast, HIV-infected children had a high proportion of predominant CDR3 lengths within both CD45RA and CD45RO T-cell subpopulations, most commonly in CD8(+) CD45RO T cells. Sharp differences in clonal dominance and size distributions were observed when cells were separated into CD45RA or CD45RO subpopulations. These differences were not apparent in unfractionated CD4(+) or CD8(+) T cells from HIV-infected subjects. Sequence analysis of predominant CDR3 lengths revealed oligoclonal expansion within individual Vbeta families. Analysis of the CDR3 length diversity within CD45RA and CD45RO T cells provides a more accurate measure of disturbances in the TCR repertoire than analysis of unfractionated CD4 and CD8 T cells. PMID- 11063506 TI - Fecal excretion of a novel human circovirus, TT virus, in healthy children. AB - The role of TT virus (TTV) as a human pathogen is unclear, as is the mode of TTV transmission. To determine the prevalence of TTV infection and the possible fecal oral route of transmission, we analyzed fecal specimens from 67 healthy, nontransfused children for TTV DNA sequences by heminested PCR, using the NG and T primer sets. The overall prevalence of TTV fecal excretion was 22.4% (15 of 67), with the T primer set (19.4%) being more sensitive than the NG primer set (10.4%). TTV prevalence based on gender or ethnicity showed no significant differences. None of seven children in the 0- to 6-month age group had detectable TTV in feces. Of three sets of siblings, two unrelated sets of twins, ages 33 and 37 months, were negative for fecal TTV DNA, while the third set of siblings, ages 99 and 35 months, was positive. The absence of TTV in the feces of children younger than 6 months and the high prevalence (40%) in children 7 to 12 months of age is consistent with age-specific acquisition of TTV infection by the nonparenteral route. TTV genotypes 1, 3, 4, and 5 were represented in our study population. TTV-positive siblings had TTV genotypes 1 and 4, suggesting unrelated environmental sources of TTV infection. This observation suggests a possible time frame for TTV acquisition in children which coincides with increased interaction with their environment and increased susceptibility to infectious agents. PMID- 11063507 TI - Evaluation of antibodies against a rubella virus neutralizing domain for determination of immune status. AB - The protective immune responses against rubella virus (RV) are related to its neutralizing epitopes, an issue that is important to consider when assessing the immune status of patients with remote infection. In the present paper, we compare the antibodies detected by a synthetic-peptide-based enzyme immunoassay (EIA) with antibodies detected by the traditional technique of hemagglutination inhibition (HIA) in patients with remote RV infection. The synthetic peptide used as an antigen (SP15) represents a neutralizing epitope that corresponds to amino acids 208 to 239 of the E1 glycoprotein. The SP15-EIA was developed, all variables that affected the assay were standardized, and the test was validated using reference sera. Serum samples (n = 129) from patients with remote RV infection were tested by HIA and SP15-EIA. Discrepant sera were assayed by MEIA (IMX/Abbot). The comparison between HIA and SP15-EIA, taking HIA as the standard methodology for determining immune status, showed that SP15-EIA is very specific and sensitive for detecting protecting antibodies (specificity, 100%; sensitivity, 98.20%). This study demonstrates that antibodies against the neutralizing domain represented by SP15 would be important in the memory response after natural infection and may be a good tool in the determination of the true immune status of patients with remote infection with regard to RV. PMID- 11063508 TI - Recombinant p51 as antigen in an immune complex transfer enzyme immunoassay of immunoglobulin G antibody to human immunodeficiency virus type 1. AB - An ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) of antibody immunoglobulin G (IgG) to human immunodeficiency virus type 1 (HIV-1) has been developed using recombinant HIV-1 reverse transcriptase (rRT) as antigen. However, some disadvantages were noted in the use of rRT as antigen: rRT was produced only with low efficiency in widely used strains of Escherichia coli using a rather long DNA fragment (3,012 bp) of the whole HIV-1 pol gene, and it was impossible to produce fusion proteins of RT for simple purification, since rRT is a heterodimer of p66 and p51. In this study, recombinant HIV-1 p51 and p66 with Ser-Ser at the N termini (Ser-Ser-rp51 and Ser-Ser-rp66) were produced in E. coli as fusion proteins with maltose binding protein containing a factor Xa site between the two proteins and were purified after digestion with factor Xa. Ser Ser-rp51 was produced in larger amounts and purified in higher yields with less polymerization than Ser-Ser-rp66. Polymerized Ser-Ser-rp66 tended to be precipitated on mercaptoacetylation for conjugation to beta-D-galactosidase (used as a label) and showed higher nonspecific and lower specific signals in an immune complex transfer enzyme immunoassay of antibody IgG to HIV-1 than Ser-Ser-rp51. The signals for serum samples of HIV-1-seropositive subjects by immune complex transfer enzyme immunoassay of antibody IgG to HIV-1 using Ser-Ser-rp51 as antigen (Y) were well correlated to those obtained using rRT as antigen (X) (log Y = 0.99 log X + 0.23; r = 0.99). Thus, the use of rp51 as antigen was advantageous over that of rp66 and rRT in an immune complex transfer enzyme immunoassay of antibody IgG to HIV-1. PMID- 11063509 TI - Rapid detection of Salmonella enterica serovar Choleraesuis in blood cultures by a dot blot enzyme-linked immunosorbent assay. AB - A dot blot enzyme-linked immunosorbent assay (ELISA) with a monoclonal antibody specific to phase1-c Salmonella was developed for the direct detection of Salmonella enterica serovar Choleraesuis in blood cultures. This system was applied to the identification of serovar Choleraesuis, and the results were compared with those obtained by a conventional biochemical method. It was revealed that all 12 samples identified to be infected with serovar Choleraesuis were positive on testing by the ELISA. In contrast, 77 samples infected with bacteria commonly isolated from the blood were not reactive by the ELISA. The calculated sensitivity and specificity of the established assay are 100%. PMID- 11063510 TI - Inhibition of human peripheral blood neutrophil respiratory burst by alcohol based venipuncture site disinfection. AB - Ethanol inhibits the respiratory burst of neutrophils. Therefore, the effects of alcohol-based skin disinfection on oxygen metabolism in neutrophils were tested using 70% ethanol or an ethanol-isopropanol-n-propanol mixture. Neutrophil respiratory burst activity as assessed fluorometrically by oxidation of 2', 7' dichlorofluorescein diacetate increased at 10 min after disinfection with 70% ethanol compared to the activity at 30 s. The increase was significant for triggering oxidative burst with formyl-peptide but not with phorbol myristate acetate. PMID- 11063511 TI - Monoclonal antibody binding to a surface-exposed epitope on Cowdria ruminantium that is conserved among eight strains. AB - Monoclonal antibodies (MAb) binding to Cowdria ruminantium elementary bodies (EB) were identified by enzyme-linked immunosorbent assay, and surface binding of one MAb (446.15) to intact EB was determined by immunofluorescence, immunogold labeling, and transmission electron microscopy. MAb 446.15 bound an antigen of approximately 43 kDa in immunoblots of eight geographically distinct strains. The MAb did not react with Ehrlichia canis antigens or uninfected bovine endothelial cell lysate and may be useful in diagnostic assays and vaccine development. PMID- 11063512 TI - Molecular confirmation of human immunodeficiency virus (HIV) type 2 in HIV seropositive subjects in south India. AB - Nested PCRs for human immunodeficiency virus type 1 (HIV-1) and HIV-2 were compared with immunoblot test results. Twelve of 13 immunoblot-positive HIV-2 samples were positive by PCR. There were five INNO-LIA (Innogenetics, Zwijnaarde, Belgium) and/or HIVBLOT 2.2 (Genelabs, Singapore) samples that tested positive for dual infection. HIV-1 PCR was positive in all samples, while HIV-2 PCR was positive in two and RIBA (Chiron Corporation, San Diego, Calif.) was positive for HIV-2 in three samples. Thus the prevalence of HIV-2 is accurately estimated by the use of immunoblotting, but that of HIV-1 and -2 dual infection may be overestimated. PMID- 11063513 TI - Learning from our international colleagues PMID- 11063514 TI - Part I: an introduction to conducting qualitative research in children with cancer. AB - Over the past decade, pediatric nurse researchers have acknowledged the need to study children's cancer illness experiences within the qualitative research framework. Support for more qualitative research is based on the belief that it will afford researchers the opportunity to get closer to understanding children's perspectives of their cancer experience. A priori theories or generalizations by the researcher are not imposed; therefore, information emerging from the research is believed to be more a reflection of the perspectives of child participants and not adult researchers. Although pediatric oncology nurses may be interested in using more qualitative methods in their research, deciding on the appropriate qualitative research design may not always be so evident, considering that the adoption of qualitative inquiry in the study of childhood cancer is in its infancy. Accordingly, the purpose of this article is to increase the reader's understanding of the use of the qualitative research paradigm in the study of children's experiences with cancer. An overview of four qualitative research designs that pediatric oncology nurse researchers may adopt is presented. Specifically, the qualitative designs of grounded theory, ethnography, phenomenology, and biography or illness narratives are examined. To facilitate discussion, each of the four designs are applied to the study of symptom experiences in children with cancer. PMID- 11063515 TI - Part II: a critical review of qualitative research related to children's experiences with cancer. AB - Using the qualitative research process to study children's experiences with cancer is being promoted because it is believed that it will afford researchers the opportunity to access children's perspectives of their cancer experiences. A detailed understanding of children's experiences with cancer will result, including a description of their feelings, wants, needs, and concerns. The information gained from qualitative research will help pediatric oncology nurses to better understand what their patients are experiencing. Although adoption of the qualitative paradigm in the study of childhood cancer is in its infancy, qualitative research findings related to children's perspectives of cancer are now slowly emerging. Accordingly, the purpose of this article is to identify some of the meaningful contributions that qualitative research has made in advancing the knowledge base of children's cancer experiences. The first section of this article discusses major themes that have evolved from use of the qualitative research process in the study of children's cancer experiences. This discussion is followed by a critique of the research with suggestions that will aid pediatric nurse researchers in conducting qualitative research when children with cancer are the research participants. PMID- 11063516 TI - Development of two instruments measuring social support for siblings of children with cancer. AB - The literature on childhood cancer provides a very limited understanding of healthy siblings' perceptions of supportive interventions during the childhood cancer experience. The purpose of this article is to discuss the development of the Nurse-Sibling Social Support Questionnaire (NSSSQ). Instrument methodology for the study involved item development, face and content validation, and internal consistency reliability as described by Hockenberry-Eaton, Manteuffel, and Bottomley (1997). Item development for the research instrument evolved from an extensive review of the literature and clinical experience of the principal investigator. Content validity of the instrument was accomplished by five experienced pediatric oncology nurses according to the methodology described by Lynn (1986). Using the content validity index, each nurse rated each item as either 4 or 5, indicating 100% agreement among experts that these items measured the concept of social support. Readability for the instrument was determined by using a computerized program. Results showed that readability was concordant with the grade school level for all items. Twenty-five school-age siblings of children with cancer and their mothers were asked to complete the questionnaire. Instrument completion was accomplished in less than 1 hour. The NSSSQ showed high internal consistencies (alpha coefficients >.90). Results indicated that siblings' perceptions of social support differed from those of their mothers. Siblings perceive emotional and instrumental support as greater in importance, whereas mothers perceive emotional and informational support as more beneficial to siblings. Support issues for siblings of children with cancer have been difficult to assess because of the lack of appropriate instruments. This study finding provides exploratory evidence to suggest that the new instrument can help measure siblings' perceptions of social support during the childhood cancer experience. PMID- 11063517 TI - Important aspects of care and assistance for children with cancer. AB - This study examined aspects of care and assistance that are important for 8- to 12-year-old children with cancer. Data were gathered through interviews with 25 children, 31 parents, and 32 nurses. Each participant was asked: "What caring aspects are important for you/your child/the child to feel cared for?" and "What help, if any, do you/your child/the child need outside the hospital?" Data were analyzed by content analysis. The following important caring aspects were identified: amusement, clinical competence, continuity, family participation, honest communication, information, participation in decision making, satisfaction of basic needs, social competence, and time. Children most frequently mentioned the importance of social competence, amusement, and satisfaction of basic needs. Parents and nurses most frequently mentioned the importance of information, social competence, and participation in decision making. The following important assistance aspects were also identified: emotional support, family life, meeting friends, practical support, rehabilitation, and school support. Two-thirds of the children did not mention that they needed any help outside the hospital. According to parents and nurses, one third of the children needed emotional support, whereas none of the children mentioned a need for this. PMID- 11063518 TI - Ethical issues: are we acting as moral agents for our patients? PMID- 11063519 TI - Roadmaps: what it's like. PMID- 11063520 TI - Principles in palliative care: an overview. PMID- 11063521 TI - Dyspnea assessment. PMID- 11063522 TI - Dyspnea treatment. PMID- 11063523 TI - Managing secretions in dying patients. PMID- 11063524 TI - Role of pulmonary rehabilitation in palliative care. PMID- 11063525 TI - Noninvasive ventilation at the end of life. PMID- 11063526 TI - Communicating with patients and their families about advance care planning and end-of-life care. PMID- 11063527 TI - Withdrawing life-sustaining treatment in the intensive care unit. PMID- 11063529 TI - Time is of the essence. PMID- 11063528 TI - Will statins unseat estrogen? PMID- 11063530 TI - The heart is connected to the... bladder? PMID- 11063531 TI - Ask the doctor. I am a 40 year old lawyer, and am generally healthy and fit. However, I get a crushing chest pain every now and then... I go to the emergency department to be sure I'm not having a heart attack. After the last visit, I had a coronary angiogram to try to see whether I had heart problems, but it showed no evidence of atherosclerosis... I am still nervous that the test might have missed something. PMID- 11063532 TI - Ask the doctor. I am trying to decide whether to buy a home blood pressure monitor. I have mild high blood pressure (my doctor has recorded some readings lately in the vicinity of 170/90 mm Hg). He is starting me on medicines now. Should I lay out the money for a monitor? PMID- 11063533 TI - Ten things you should know about heart disease. PMID- 11063534 TI - Alternative medicine. Willow bark for low back pain. PMID- 11063535 TI - Hearing. Age, hearing loss, and hearing aids. PMID- 11063536 TI - Tips about having surgery. PMID- 11063537 TI - By the way, doctor... I recently had a hacking cough, and my doctor told me that I had bronchitis. He told me I didn't need antibiotics. is this some managed-care attempt to save money at my expense? PMID- 11063538 TI - SSRIs: Prozac and company - part II. PMID- 11063539 TI - Insight in psychosis. PMID- 11063540 TI - Old drug, new label. PMID- 11063541 TI - Environment and drug abuse. PMID- 11063542 TI - Harbingers of schizophrenia. PMID- 11063543 TI - What are the uses and dangers of kava? PMID- 11063544 TI - Colon cancer. PMID- 11063545 TI - The write stuff. PMID- 11063546 TI - Carbon monoxide: a silent killer. PMID- 11063547 TI - On call. I've always been completely healthy. At my last check-up, my cholesterol was 295. I'm worried, but my doctor tells me I don't need treatment. What do you suggest? PMID- 11063548 TI - St. John's wort for depression. PMID- 11063549 TI - Women and moderate drinking: health in the balance. PMID- 11063550 TI - Breast cancer update. Part II: diagnosis and early treatment. PMID- 11063551 TI - By the way, doctor. What are the differences between vaginal and abdominal hysterectomy? What are the advantages of each? Also, what are the reasons to have the ovaries removed at the same time? PMID- 11063552 TI - By the way, doctor. I am a postmenopausal woman (age 56) with serious varicose veins. I would appreciate information on exercises to increase my lower body strength without making my varicose veins worse than they already are. PMID- 11063553 TI - New hepatitis viruses. AB - Three new hepatitis viruses are reviewed; Hepatitis E Virus (HEV), Hepatitis G Virus (HGV), and Transfusion Transmitted Virus (TTV). Only HEV has been clearly associated with liver damage. It is transmitted by the fecal-oral route and tends to occur in poor socio-economic conditions. Acute disease is diagnosed by the presence of anti-HEV IgM antibodies in the serum. Hepatitis G virus and TTV are transmitted parenterally. HGV is the same agent as GBV-C. Although it is hepatotropic, and high levels of viremia may occur, pathogenicity to the liver has not been proven. TTV may also be transmitted by the fecal-oral route. It is abundant in liver tissue but, like HGV, pathogenicity has not been proven. PMID- 11063554 TI - In vitro antimicrobial activity of piperacillin/tazobactam in comparison with other broad-spectrum beta-lactams. AB - Combining tazobactam, a beta-lactamase inhibitor, with the ureidopenicillin, piperacillin, successfully restores the activity of piperacillin against beta lactamase producing bacteria. Thus, piperacillin/tazobactam is highly active against most clinically important species of Gram-negative and Gram-positive bacteria, including anaerobes. We evaluated the in vitro activity of piperacillin/tazobactam against clinical isolates from a tertiary university hospital located in Sao Paulo, Brazil. Its activity was compared to that of ticarcillin/clavulanic acid, ampicillin/sulbactam, ceftazidime, ceftriaxone, cefotaxime, cefoxitin, aztreonam, and imipenem against 820 isolates (608 Gram negative and 212 Gram-positive) collected from hospitalized patients in 1999. The most frequent species tested were Pseudomonas aeruginosa (168/20%), Escherichia coli (139/17%), Acinetobacter spp. (131/16%), and Staphylococcus aureus (76/9%). Of the isolates studied, 30% were from the bloodstream, 16% from the lower respiratory tract, and 11% from surgical wounds or soft tissue. The isolates were susceptibility tested by the broth microdilution method according to NCCLS procedures. The isolates tested were highly resistant to most antimicrobial agents evaluated. Imipenem resistance was not verified among Enterobacteriaceae, and piperacillin/tazobactam was the second most active beta-lactams against this group of bacteria (80.0% susceptibility). Extended-spectrum beta-lactamase production was very high among E. coli (approximately 20%) and Klebsiella pneumoniae (approximately 40%). Imipenem was uniformly active against these species (100% susceptibility) and piperacillin/tazobactam was the second most active compound inhibiting 84.4% of isolates. Pseudomonas aeruginosa was highly resistant to all beta-lactams evaluated and piperacillin/tazobactam was the most active compound against this species. Our results demonstrate an extremely high level of antimicrobial resistance in the hospital evaluated, especially among non enteric Gram-negative bacilli. Due to this high level of resistance, piperacillin/tazobactam represents an important contribution to the treatment of nosocomial infections. PMID- 11063555 TI - Evaluation of the in vitro activity of 9 antimicrobials against bacterial strains isolated from patients in intensive care units in brazil: MYSTIC Antimicrobial Surveillance Program. AB - Multi-resistant bacterial strains are increasingly prevalent in hospital environments. Bacterial resistance is an important problem, especially for practitioners in intensive care units (ICUs) because of the selective pressure on the prevalent bacteria in these environments. The MYSTIC (Meropenem Yearly Susceptibility Test Information Collection) study has been monitoring the performance of carbapenems and other antibiotics in different hospitals for at least 3 years. The in vitro activities of meropenem, imipenem, ceftazidime, cefepime, cefotaxime, ciprofloxacin, piperacilin/tazobactam, gentamicin, and tobramycin were compared against 452 recent clinical aerobic isolates. The isolates consisted of 19 species of Gram-negative bacteria (n=290) including K. pneumoniae (n=49), E. coli (n=48), A. baumannii (n=47), Enterobacter spp. (n=41), and P. aeruginosa (n=33) and 9 species of Gram-positive bacteria (n=162) including Staphylococcus aureus (n=63), Enterococcus faecalis (n=22), Streptococcus pneumoniae (n=22) and coagulase negative Staphylococci (n=21). All isolates were collected from ICU patients. Minimal inhibitory concentrations (MICs) were determined by Etest methodology, using standardized and controlled procedures. Meropenem and imipenem showed the lowest MIC(90) for all species tested. Gram-negative isolates showed the following overall resistance percentages to the other 7 drugs: tobramycin (43.1%), cefotaxime (38.6%), gentamicin (34.1%), ceftazidime (31.7%), ciprofloxacin (25.5%), piperacillin/tazobactam (26.9%), and cefepime (18.6%). Carbapenems were the most active drugs overall and only P. aeruginosa presented some degree of resistance (18.2%). We also evaluated the production of extended spectrum beta-lactamase (ESBL) among all Enterobacteriaceae members (n=176) by Etest/ESBL strip. ESBL production was detected in 51 strains (29.0%). Among them, Klebsiella pneumoniae was the most prevalent at 59.2%, followed by Enterobacter spp. (19.5%) and E. coli (14.6%). The high level of resistance against several antimicrobials and the alarming rate of ESBL production may restrict therapeutic choice to the carbapenems in this selected group of patients. PMID- 11063556 TI - Prevalence of antimicrobial resistance among respiratory tract isolates in Latin America: results from SENTRY antimicrobial surveillance program (1997-98). AB - One thousand seventy-three bacterial isolates were collected from patients with community acquired respiratory tract infections (CARTI) in 11 Latin American centers (7 countries) during 1997 and 1998. They were tested against numerous antimicrobial agents by the reference broth microdilution method as part of the ongoing multinational SENTRY Antimicrobial Surveillance Program. Among Streptococcus pneumoniae (553 isolates), approximately 61% were susceptible to penicillin. There was a great variation of the penicillin susceptibility rates among participating countries. The highest susceptibility rates were found in Argentina (76.7%) and Brazil (71.9%), while the lowest rate of penicillin susceptibility was detected in Mexico (33.3%). High level resistance to penicillin and resistance to cefotaxime were observed in nearly 10% of the isolates. The newer quinolones, levofloxacin (MIC(90) 2 microg/mL) and gatifloxacin (MIC90 0.5 microg/mL), were active against 100% of the isolates tested. Among the other non-beta-lactams drugs tested, the rank order of susceptibility against the pneumococci was: chloramphenicol (93.9%)>clindamycin (93.2%)> azithromycin (89.1%) > clarithromycin (88.7%)>tetracycline (78.5%)> trimethoprim/sulfamethoxazole (55.7%). The percentage of Haemophilus influenzae (361 isolates) isolates resistant to amoxicillin was 12. 7% (beta-lactamase positive). Among Moraxella catarrhalis (159 isolates) isolates, only 8.2% were susceptible. Clavulanic acid restored the activity of amoxicillin against both species. Trimethoprim/sulfamethoxazole was active against only 59.5% of H. influenzae, while susceptibility to this compound among M. catarrhalis was 96.1%. All other compounds tested were active against>95% of H. influenzae and M. catarrhalis isolates. These species were susceptible to levofloxacin (MIC90 < or = 0.5 microg/mL for both) and gatifloxacin (MIC90 < or = 0.03 microg/mL for both) with very low MICs. Our results indicate that penicillin resistance rates are particularly high among pneumococci in some countries. The newer fluoroquinolones show an excellent potency and spectrum against pathogens causing community acquired respiratory infections in Latin America. PMID- 11063557 TI - Disseminated cutaneous histoplasmosis and AIDS: case report. AB - Histoplasmosis is caused by the dimorphic fungus Histoplasma capsulatum. It manifests by the presence of fever as the only symptom in most individuals. The disease may present as self-limited pneumonia, or as an hematogenous widespread fungal infection with a potentially fatal outcome in elderly individuals and people with compromised T-cell mediated immunity. Here, we report a case of disseminated cutaneous histoplasmosis in a patient with AIDS. The patient was a 33 year old male homosexual, intravenous drug user, who had been diagnosed with HIV infection 5 years earlier. He was in good health, but had erythematous papules and pustules in the skin of the scalp, face, back, thighs, abdomen, palms, and soles. He was placed on anti-retroviral therapy, fluconazole for mucosal candidiasis, trimethoprim/sulfamethoxazole for pneumocystis prophylaxis, and antibiotics for the skin pustules. The skin lesions improved remarkably within 14 days. He was discharged and soon lost to follow-up. After his discharge, skin biopsy and fungal culture results revealed H. capsulatum. He was seen again 1 year later. The interim history revealed that he had taken fluconazole 100 mg/day for 1 month and fluconazole 150 mg/week for 7 months. He had not continued anti-retroviral therapy, nor taken other antifungal drugs. The clinical evolution of the disease was exceptional in that there was disappearance of all the skin lesions attributed to histoplasmosis with fluconazole. Although itraconazole remains the drug of choice for histoplasmosis. Cutaneous histoplasmosis should be considered in the differential diagnosis of atypical cutaneous lesions in individuals infected with HIV. PMID- 11063558 TI - A case of keratoconjunctivitis due to Ewingella americana and a review of unusual organisms causing external eye infections. AB - We report the isolation of Ewingella americana from the conjunctivae of a 38 year old female physician with keratoconjunctivitis associated with the use of soft contact lens. The patient was treated successfully with topical ciprofloxacin. The source of the infection remains unknown. All contact lens cleaning materials used by the patient were sterile. Since the patient was a physician, and this organism has been recorded as a cause of nosocomial infections, we checked whether cases of Ewingella americana had been reported, but none were identified. We have identified 39 bacterial species, 27 fungi, 4 viruses, 7 protozoa, 4 helminths, and 2 arthropods which rarely have been associated with keratitis or conjunctivitis. Infectious diseases specialists and ophthalmologists must be aware of the many different causes of this illness, including Ewingella americana. This organism is a rare bacterial cause of keratoconjunctivitis not previously reported in Brazil. It should be added to the list of unusual cases of external eye infections. PMID- 11063559 TI - Etest compared to broth microdilution: discrepant results when testing macrolides against Streptococcus pneumoniae indicate a need for better clinical and serum level/MIC correlation. PMID- 11063560 TI - DASH diet lowers homocysteine levels. PMID- 11063561 TI - Ask the doctor. My blood pressure is 180/80 mm Hg. My doctor tells me this a common problem for people in their 70s, like me. The problem is that whenever I try medicines at doses high enough to get my top number under 140 mm Hg (which I understand to be the goal), I get exhausted or have other side effects. Do I really need to worry about my blood pressure when the bottom number is so good? PMID- 11063562 TI - By the way, doctor... A local hospital is advertising that I can get a CT scan of my entire body for $700. If it's negative, they say that shows that I don't have cancer. My insurance company won't cover this, but it seems like it might be worth the money if one of these scans can find something. Do you think it's a good idea? PMID- 11063564 TI - How effective is child psychotherapy? PMID- 11063563 TI - Brain signals of dementia? PMID- 11063565 TI - Ion channels, permeation, and electrostatics: insight into the function of KcsA. PMID- 11063566 TI - Structure and mechanism of peptide methionine sulfoxide reductase, an "anti oxidation" enzyme. AB - Peptide methionine sulfoxide reductase (MsrA) reverses oxidative damage to both free methionine and methionine within proteins. As such, it helps protect the host organism against stochastic damage that can contribute to cell death. The structure of bovine MsrA has been determined in two different modifications, both of which provide different insights into the biology of the protein. There are three cysteine residues located in the vicinity of the active site. Conformational changes in a glycine-rich C-terminal tail appear to allow all three thiols to come together and to participate in catalysis. The structures support a unique, thiol-disulfide exchange mechanism that relies upon an essential cysteine as a nucleophile and additional conserved residues that interact with the oxygen atom of the sulfoxide moiety. PMID- 11063567 TI - Structure of a murine cytoplasmic serine hydroxymethyltransferase quinonoid ternary complex: evidence for asymmetric obligate dimers. AB - Serine hydroxymethyltransferase (SHMT) is a pyridoxal phosphate-dependent enzyme that catalyzes the reversible conversion of serine and tetrahydrofolate to glycine and methylenetetrahydrofolate. This reaction generates single carbon units for purine, thymidine, and methionine biosynthesis. The enzyme is a homotetramer comprising two obligate dimers and four pyridoxal phosphate-bound active sites. The mammalian enzyme is present in cells in both catalytically active and inactive forms. The inactive form is a ternary complex that results from the binding of glycine and 5-formyltetrahydrofolate polyglutamate, a slow tight-binding inhibitor. The crystal structure of a close analogue of the inactive form of murine cytoplasmic SHMT (cSHMT), lacking only the polyglutamate tail of the inhibitor, has been determined to 2.9 A resolution. This first structure of a ligand-bound mammalian SHMT allows identification of amino acid residues involved in substrate binding and catalysis. It also reveals that the two obligate dimers making up a tetramer are not equivalent; one can be described as "tight-binding" and the other as "loose-binding" for folate. Both active sites of the tight-binding dimer are occupied by 5-formyltetrahydrofolate (5 formylTHF), whose N5-formyl carbon is within 4 A of the glycine alpha-carbon of the glycine-pyridoxal phosphate complex; the complex appears to be primarily in its quinonoid form. In the loose-binding dimer, 5-formylTHF is present in only one of the active sites, and its N5-formyl carbon is 5 A from the glycine alpha carbon. The pyridoxal phosphates appear to be primarily present as geminal diamine complexes, with bonds to both glycine and the active site lysine. This structure suggests that only two of the four catalytic sites on SHMT are catalytically competent and that the cSHMT-glycine-5-formylTHF ternary complex is an intermediate state analogue of the catalytic complex associated with serine and glycine interconversion. PMID- 11063568 TI - Reaction intermediate analogues for mandelate racemase: interaction between Asn 197 and the alpha-hydroxyl of the substrate promotes catalysis. AB - Mandelate racemase (MR) catalyzes the interconversion of the enantiomers of mandelic acid, stabilizing the altered substrate in the transition state by 26 kcal/mol relative to the substrate in the ground state. To understand the origins of this binding discrimination, carboxylate-, phosphonate-, and hydroxamate containing substrate and intermediate analogues were examined for their ability to inhibit MR. Comparison of the competitive inhibition constants revealed that an alpha-hydroxyl function is required for recognition of the ligand as an intermediate analogue. Two intermediate analogues, alpha-hydroxybenzylphosphonate (alpha-HBP) and benzohydroxamate, were bound with affinities approximately 100 fold greater than that observed for the substrate. Furthermore, MR bound alpha HBP enantioselectively, displaying a 35-fold higher affinity for the (S) enantiomer relative to the (R)-enantiomer. In the X-ray structure of mandelate racemase [Landro, J. A., Gerlt, J. A., Kozarich, J. W., Koo, C. W., Shah, V. J., Kenyon, G. L., Neidhart, D. J., Fujita, J., and Petsko, G. A. (1994) Biochemistry 33, 635-643], the alpha-hydroxyl function of the competitive inhibitor (S) atrolactate is within hydrogen bonding distance of Asn 197. To demonstrate the importance of the alpha-hydroxyl function in intermediate binding, the N197A mutant was constructed. The values of k(cat) for N197A were reduced 30-fold for (R)-mandelate and 179-fold for (S)-mandelate relative to wild-type MR; the values of k(cat)/K(m) were reduced 208-fold for (R)-mandelate and 556-fold for (S) mandelate. N197A shows only a 3.5-fold reduction in its affinity for the substrate analogue (R)-atrolactate but a 51- and 18-fold reduction in affinity for alpha-HBP and benzohydroxamate, respectively. Thus, interaction between Asn 197 and the substrate's alpha-hydroxyl function provides approximately 3.5 kcal/mol of transition-state stabilization free energy to differentially stabilize the transition state relative to the ground state. PMID- 11063569 TI - A key role in catalysis for His89 of adenylosuccinate lyase of Bacillus subtilis. AB - Adenylosuccinate lyase of Bacillus subtilis is a tetrameric enzyme which catalyzes the cleavage of adenylosuccinate to AMP and fumarate. We have mutated His(89), one of three conserved histidines, to Gln, Ala, Glu, and Arg. The enzymes were expressed in Escherichia coli and purified to homogeneity. As compared to a specific activity of 1. 56 micromol of adenylosuccinate converted/min/mg protein for wild-type enzyme, the mutant enzymes exhibit specific activities of 0.0225, 0.0036, 0.0036, and 0.0009 for H89Q, H89A, H89E, and H89R, respectively. Circular dichroism and FPLC gel filtration reveal that mutant enzymes have a similar conformation and oligomeric state to that of wild type enzyme. In H89Q, the K(M) for adenylosuccinate increases slightly to 2.5 fold that of wild-type, the K(M) for fumarate is elevated 3.3-fold, and the K(M) for AMP is 13 times higher than that observed in wild-type enzyme. The catalytic efficiency of the H89Q enzyme is compromised, with k(cat)/K(M) reduced 174-fold in the direction of AMP formation. These data suggest that His(89) plays a role in both the binding of the AMP portion of the substrate and in correctly orienting the substrate for catalysis. Incubation of H89Q with inactive H141Q enzyme [Lee, T. T., Worby, C., Bao, Z.-Q., Dixon, J. E., and Colman, R. F. (1999) Biochemistry 38, 22-32] leads to a 30-fold increase in activity. This intersubunit complementation indicates that His(89) and His(141) from different subunits participate in the active site and that both are required for catalysis. PMID- 11063570 TI - Localization of the Cl-/HCO3- anion exchanger binding site to the amino-terminal region of carbonic anhydrase II. AB - Human carbonic anhydrase II (CAII) possesses a binding site for an acidic motif (D887ADD) within the carboxyl-terminal region (Ct) of the human erythrocyte chloride/bicarbonate anion exchanger, AE1. In this study, the amino acid sequence comprising this AE1 binding site was localized to the first 17 residues of CAII, which form a basic patch on the surface of the protein. Truncation of the amino terminal of CAII by five residues resulted in a 3-fold reduction in the apparent affinity of the interaction with a GST fusion protein of the Ct of AE1 (GST-Ct) measured by a sensitive microtiter plate binding assay. Further amino-terminal truncation of CAII by 17 or 24 residues caused a loss of binding. The homologous isoform CAI does not bind AE1, despite having 60% sequence identity to CAII. One major difference between the two CA isoforms, within the amino-terminal region, is a high content of histidine residues in CAII (His3, -4, -10, -15, -17) not found in CAI. Mutation of pairs of these histidines (and one lysine) in CAII to the analogous residues in CAI (H3P/H4D or K9D/H10K or H15Q/H17S), or combinations of these various double mutants, did not greatly affect binding between GST-Ct and the mutant CAII. However, when all six of the targeted CAII residues were mutated to the corresponding sequence in CAI, binding of GST-Ct was lost. These results indicate that the AE1 binding site is located within the first 17 residues of CAII, and that the interaction is mediated by electrostatic interactions involving histidine and/or lysine residues. Further specificity for the interaction of AE1 and CAII is provided by a conserved leucine residue (L886) in AE1 that, when mutated to alanine, resulted in loss of GST-Ct binding to immobilized CAII. The binding of the basic amino-terminal region of CAII to an acidic Ct in AE1 provides a structural basis for linking bicarbonate transport across the cell membrane to intracellular bicarbonate metabolism. PMID- 11063571 TI - Removal of the four C-terminal glycine-rich repeats enhances the thermostability and substrate binding affinity of barley beta-amylase. AB - Barley beta-amylase undergoes proteolytic cleavage in the C-terminal region after germination. The implication of the cleavage in the enzyme's characteristics is unclear. With purified native beta-amylases from both mature barley grain and germinated barley, we found that the beta-amylase from germinated barley had significantly higher thermostability and substrate binding affinity for starch than that from mature barley grain. To better understand the effect of the proteolytic cleavage on the enzyme's thermostability and substrate binding affinity for starch, recombinant barley beta-amylases with specific deletions at the C-terminal tail were generated. The complete deletion of the four C-terminal glycine-rich repeats significantly increased the enzyme's thermostability, but an incomplete deletion with one repeat remaining did not change the thermostability. Although different C-terminal deletions affect the thermostability differently, they all increased the enzyme's affinity for starch. The possible reasons for the increased thermostability and substrate binding affinity, due to the removal of the four C-terminal glycine-rich repeats, are discussed in terms of the three dimensional structure of beta-amylase. PMID- 11063572 TI - Dynamics of the metallo-beta-lactamase from Bacteroides fragilis in the presence and absence of a tight-binding inhibitor. AB - A significant determinant for the broad substrate specificity of the metallo-beta lactamases from Bacteroides fragilis and other similar organisms is the presence of a plastic substrate binding site that is nevertheless capable of tight substrate binding in the Michaelis complex. To achieve these two competing ends, the molecule apparently employs a flexible flap that closes over the active site in the presence of substrate. These characteristics imply that dynamic changes are an important component of the mechanism of action of these enzymes. The backbone and tryptophan side chain dynamics of the metallo-beta-lactamase from B. fragilis have been examined using (15)N NMR relaxation measurements. Two states of the protein were examined, in the presence and absence of a tight-binding inhibitor. Relaxation measurements were analyzed by the model-free method. Overall, the metallo-beta-lactamase molecule is rigid and shows little flexibility except in loops. The flexibility of the loop that covers the active site is not unusually great as compared to the other loops of the protein. Local motion on a picosecond time scale was found to be very similar throughout the protein in the presence and absence of the inhibitor, but a significant difference was observed in the motions on a nanosecond time scale (tau(e)). Large amplitude motions with a time constant of about 1.3 ns were observed for the flexible flap region (residues 45-55) in the absence of the inhibitor. These motions were completely damped out in the presence of the inhibitor. In addition, the motion of a tryptophan side chain at the tip of the beta-hairpin of the flap shows a very significant difference in motion on the ps time scale. These results indicate that the motions of the polypeptide chain in the flap region can be invoked to explain both the wide substrate specificity (the free form has considerable amplitude of motion in this region) and the catalytic efficiency of the metallo-beta-lactamase (the motions are damped out when the inhibitor and by implication a substrate binds in the active site). PMID- 11063573 TI - Evaluation of the utility of NMR structures determined from minimal NOE-based restraints for structure-based drug design, using MMP-1 as an example. AB - The application of deuterium labeling and residual dipolar coupling constants in combination with other structural information has demonstrated the potential for significantly expanding the range of viable protein targets for structural analysis by NMR. A previous study by Clore et al. [(1999) J. Am. Chem. Soc. 121, 6513-6514] demonstrated that a significant improvement in the overall protein structure occurs with the combination of residual dipolar coupling constants and minimal tertiary long-range distance restraints. The analysis of NMR protein structures determined with minimal structural information is extended with a particular interest in the utility of these structures for a structure-based drug design program. As an example, the catalytic fragment of human fibroblast collagenase (MMP-1) was used to follow the effect of minimal restraint sets on the protein structure and its utility in drug design with a particular interest in the effect on the active site conformation. An MMP-1 structure that was calculated with the maximal number of restraints attainable with the constraint of a deuterated protein was shown to be very similar to a high-quality MMP-1 structure that was calculated from a complete set of restraints. The superposition of the active site backbone atoms for the high-quality and minimal restraint MMP-1 structures yielded an rmsd of 0.68 A where the size and shape of the S1' pocket are nearly identical. Additionally, an MMP-1-CGS-27023A complex based on a minimal set of NOE-based restraints reliably reproduced the structure of the complex, establishing the usefulness of the structures for drug design. PMID- 11063574 TI - Dimer formation through domain swapping in the crystal structure of the Grb2-SH2 Ac-pYVNV complex. AB - Src homology 2 (SH2) domains are key modules in intracellular signal transduction. They link activated cell surface receptors to downstream targets by binding to phosphotyrosine-containing sequence motifs. The crystal structure of a Grb2-SH2 domain-phosphopeptide complex was determined at 2.4 A resolution. The asymmetric unit contains four polypeptide chains. There is an unexpected domain swap so that individual chains do not adopt a closed SH2 fold. Instead, reorganization of the EF loop leads to an open, nonglobular fold, which associates with an equivalent partner to generate an intertwined dimer. As in previously reported crystal structures of canonical Grb2-SH2 domain-peptide complexes, each of the four hybrid SH2 domains in the two domain-swapped dimers binds the phosphopeptide in a type I beta-turn conformation. This report is the first to describe domain swapping for an SH2 domain. While in vivo evidence of dimerization of Grb2 exists, our SH2 dimer is metastable and a physiological role of this new form of dimer formation remains to be demonstrated. PMID- 11063575 TI - Use of the parallax-quench method to determine the position of the active-site loop of cholesterol oxidase in lipid bilayers. AB - To elucidate the cholesterol oxidase-membrane bilayer interaction, a cysteine was introduced into the active site lid at position-81 using the Brevibacterium enzyme. To eliminate the possibility of labeling native cysteine, the single cysteine in the wild-type enzyme was mutated to a serine without any change in activity. The loop-cysteine mutant was then labeled with acrylodan, an environment-sensitive fluorescence probe. The fluorescence increased and blue shifted upon binding to lipid vesicles, consistent with a change into a more hydrophobic, i.e., lipid, environment. This acrylodan-labeled cholesterol oxidase was used to explore the pH, ionic strength, and headgroup dependence of binding. Between pH 6 and 10, there was no significant change in binding affinity. Incorporation of anionic lipids (phosphatidylserine) into the vesicles did not increase the binding affinity nor did altering the ionic strength. These experiments suggested that the interactions are primarily driven by hydrophobic effects not ionic effects. Using vesicles doped with either 5-doxyl phosphatidylcholine, 10-doxyl phosphatidylcholine, or phosphatidyl-tempocholine, quenching of acrylodan fluorescence was observed upon binding. Using the parallax method of London [Chattopadhyay, A., and London, E. (1987) Biochemistry 26, 39 45], the acrylodan ring is calculated to be 8.1 +/- 2.5 A from the center of the lipid bilayer. Modeling the acrylodan-cysteine residue as an extended chain suggests that the backbone of the loop does not penetrate into the lipid bilayer but interacts with the headgroups, i.e., the choline. These results demonstrate that cholesterol oxidase interacts directly with the lipid bilayer and sits on the surface of the membrane. PMID- 11063576 TI - Modified nucleoside dependent Watson-Crick and wobble codon binding by tRNALysUUU species. AB - Nucleoside modifications are important to the structure of all tRNAs and are critical to the function of some tRNA species. The transcript of human tRNA(Lys3)(UUU) with a UUU anticodon, and the corresponding anticodon stem and loop domain (ASL(Lys3)(UUU)), are unable to bind to poly-A programmed ribosomes. To determine if specific anticodon domain modified nucleosides of tRNA(Lys) species would restore ribosomal binding and also affect thermal stability, we chemically synthesized ASL(Lys) heptadecamers and site-specifically incorporated the anticodon domain modified nucleosides pseudouridine (Psi(39)), 5 methylaminomethyluridine (mnm(5)U(34)) and N6-threonylcarbamoyl-adenosine (t(6)A(37)). Incorporation of t(6)A(37) and mnm(5)U(34) contributed structure to the anticodon loop, apparent by increases in DeltaS, and significantly enhanced the ability of ASL(Lys3)(UUU) to bind poly-A programmed ribosomes. Neither ASL(Lys3)(UUU)-t(6)A(37) nor ASL(Lys3)(UUU)-mnm(5)U(34) bound AAG programmed ribosomes. Only the presence of both t(6)A(37) and mnm(5)U(34) enabled ASL(Lys3)(UUU) to bind AAG programmed ribosomes, as well as increased its affinity for poly-A programmed ribosomes to the level of native Escherichia coli tRNA(Lys). The completely unmodified anticodon stem and loop of human tRNA(Lys1,2)(CUU) with a wobble position-34 C bound AAG, but did not wobble to AAA, even when the ASL was modified with t(6)A(37). The data suggest that tRNA(Lys)(UUU) species require anticodon domain modifications in the loop to impart an ordered structure to the anticodon for ribosomal binding to AAA and require a combination of modified nucleosides to bind AAG. PMID- 11063577 TI - Functional anticodon architecture of human tRNALys3 includes disruption of intraloop hydrogen bonding by the naturally occurring amino acid modification, t6A. AB - The structure of the human tRNA(Lys3) anticodon stem and loop domain (ASL(Lys3)) provides evidence of the physicochemical contributions of N6 threonylcarbamoyladenosine (t(6)A(37)) to tRNA(Lys3) functions. The t(6)A(37) modified anticodon stem and loop domain of tRNA(Lys3)(UUU) (ASL(Lys3)(UUU)- t(6)A(37)) with a UUU anticodon is bound by the appropriately programmed ribosomes, but the unmodified ASL(Lys3)(UUU) is not [Yarian, C., Marszalek, M., Sochacka, E., Malkiewicz, A., Guenther, R., Miskiewicz, A., and Agris, P. F., Biochemistry 39, 13390-13395]. The structure, determined to an average rmsd of 1.57 +/- 0.33 A (relative to the mean structure) by NMR spectroscopy and restrained molecular dynamics, is the first reported of an RNA in which a naturally occurring hypermodified nucleoside was introduced by automated chemical synthesis. The ASL(Lys3)(UUU)-t(6)A(37) loop is significantly different than that of the unmodified ASL(Lys3)(UUU), although the five canonical base pairs of both ASL(Lys3)(UUU) stems are in the standard A-form of helical RNA. t(6)A(37), 3' adjacent to the anticodon, adopts the form of a tricyclic nucleoside with an intraresidue H-bond and enhances base stacking on the 3'-side of the anticodon loop. Critically important to ribosome binding, incorporation of the modification negates formation of an intraloop U(33).A(37) base pair that is observed in the unmodified ASL(Lys3)(UUU). The anticodon wobble position U(34) nucleobase in ASL(Lys3)(UUU)-t(6)A(37) is significantly displaced from its position in the unmodified ASL and directed away from the codon-binding face of the loop resulting in only two anticodon bases for codon binding. This conformation is one explanation for ASL(Lys3)(UUU) tendency to prematurely terminate translation and 1 frame shift. At the pH 5.6 conditions of our structure determination, A(38) is protonated and positively charged in ASL(Lys3)(UUU)-t(6)A(37) and the unmodified ASL(Lys3)(UUU). The ionized carboxylic acid moiety of t(6)A(37) possibly neutralizes the positive charge of A(+)(38). The protonated A(+)(38) can base pair with C(32), but t(6)A(37) may weaken the interaction through steric interference. From these results, we conclude that ribosome binding cannot simply be an induced fit of the anticodon stem and loop, otherwise the unmodified ASL(Lys3)(UUU) would bind as well as ASL(Lys3)(UUU)-t(6)A(37). t(6)A(37) and other position 37 modifications produce the open, structured loop required for ribosomal binding. PMID- 11063578 TI - Topology of yeast RNA polymerase II subunits in transcription elongation complexes studied by photoaffinity cross-linking. AB - The subunits of Saccharomyces cerevisiae RNA polymerase II (RNAP II) in proximity to the DNA during transcription elongation have been identified by photoaffinity cross-linking. In the absence of transcription factors, RNAP II will transcribe a double-stranded DNA fragment containing a 3'-extension of deoxycytidines, a "tailed template". We designed a DNA template allowing the RNAP to transcribe 76 bases before it was stalled by omission of CTP in the transcription reaction. This stall site oriented the RNAP on the DNA template and allowed us to map the RNAP subunits along the DNA. The DNA analogue 5-[N-(p-azidobenzoyl)-3-aminoallyl] dUTP (N(3)RdUTP) [Bartholomew, B., Kassavetis, G. A., Braun, B. R., and Geiduschek, E. P. (1990) EMBO J. 9, 2197-205] was synthesized and enzymatically incorporated into the DNA at specified positions upstream or downstream of the stall site, in either the template or nontemplate strand of the DNA. Radioactive nucleotides were positioned beside the photoactivatable nucleotides, and cross linking by brief ultraviolet irradiation transferred the radioactive tag from the DNA onto the RNAP subunits. In addition to N(3)RdUTP, which has a photoreactive azido group 9 A from the uridine base, we used the photoaffinity cross-linker 5N(3)dUTP with an azido group directly on the uridine ring to identify the RNAP II subunits closest to the DNA at positions where multiple subunits cross-linked. In cross-linking reactions dependent on transcription, RPB1, RPB2, and RPB5 were cross-linked with N(3)RdUTP. With 5N(3)dUTP, only RPB1 and RPB2 were cross linked. Under certain circumstances, RPB3, RPB4, and RPB7 were cross-linked. From the information obtained in this topological study, we developed a model of yeast RNAP II in a transcription elongation complex. PMID- 11063579 TI - Structural characterization of a guanine-quadruplex ligand complex. AB - The inhibition of telomerase by molecules such as disubstituted amidoanthraquinones is believed to be due to their stabilization of guanine quadruplex complexes. The characterization is reported of a complex with the intermolecular parallel quadruplex formed from the sequence TGGGGT and a 1,4-bis piperidino amidoanthraquinone. Crystals obtained did not give single-crystal diffraction; the fiber-like pattern has been interpreted in terms of a repeating unit with four guanine-quartets and two stacked/intercalated ligand molecules. The two categories of possible structures for the complex consistent with this interpretation have been examined by molecular dynamics simulations, with fully solvated environments and 1000 ps simulation times. The two central guanine quartets in the intercalation model rapidly became highly distorted, whereas the two types of models with ligand stacked externally on the ends of the quadruplex remained very stable. It was concluded that the externally bound ligand complexes best represent the structure of this quadruplex complex, in agreement with earlier NMR results on related systems. PMID- 11063580 TI - Functional dissection of the dimerization and enzymatic activities of Escherichia coli nitrogen regulator II and their regulation by the PII protein. AB - The dimeric two-component system transmitter protein NRII (NtrB) of Escherichia coli, product of glnL (ntrB), controls transcription of nitrogen-regulated genes by catalyzing the phosphorylation and dephosphorylation of the transcription factor NRI (NtrC). Previous studies showed that the PII signal transduction protein inhibits the kinase activity of NRII and activates its phosphatase activity. We observed that PII greatly stimulated the NRII phosphatase activity under conditions where the cleavage of ATP was prevented, indicating that the phosphatase activity did not result simply from prevention of the antagonistic NRII kinase activity by PII. Rather, PII was an activator of the phosphatase activity. To study this regulation, we examined the dimerization and enzymatic activities of NRII and various polypeptides derived from NRII, and their regulation by PII. Our results were consistent with the hypothesis that NRII consists of three domains: an N-terminal domain found only in NRII proteins and two domains formed by the conserved transmitter module of NRII, the phosphotransferase/phosphatase/dimerization (central) domain and the kinase domain. All three domains were involved in regulating the kinase and phosphatase activities of NRII. The N-terminal domain was involved in intramolecular signal transduction, and controlled access to the NRII active site for the isolated dimeric central domain added in trans. The central domain was responsible for dimerization and the phosphotransferase and phosphatase activities of NRII, but the latter activity was weak in the isolated domain and was not regulated by PII. The C-terminal kinase domain was responsible for the kinase activity. The PII protein appeared to interact with the isolated transmitter module of NRII, and not with the N-terminal domain as previously thought, since PII dramatically increased the stoichiometry of autophosphorylation of the isolated transmitter module. However, the phosphatase activity of the transmitter module of NRII was low even in the presence of PII, suggesting that the N-terminal domain was necessary for the central domain to assume the conformation necessary for potent phosphatase activity. Also, PII significantly reduced the rate of transphosphorylation of the isolated central domain by the isolated kinase domain, suggesting that PII interacts directly with the kinase domain. We hypothesize that the binding of PII to the kinase domain of NRII results in an altered conformation that is transmitted to the central and N-terminal domains; this causes the central domain to assume the conformation with potent phosphatase activity. PMID- 11063581 TI - The Escherichia coli PII signal transduction protein regulates the activities of the two-component system transmitter protein NRII by direct interaction with the kinase domain of the transmitter module. AB - The PII signal transduction protein regulates the transcription of nitrogen regulated genes by controlling the kinase and phosphatase activities of NRII. We used a cross-linking approach to study the interaction of the T-loop of the PII protein with NRII. Cross-linking of PII to NRII required ATP and 2-ketoglutarate, allosteric effectors known to control PII activity, and was not affected by the presence of excess nonspecific proteins such as bovine serum albumin. The purified cross-linked species appeared to consist mainly of PII trimers in which one of the three subunits was cross-linked to a single subunit of the NRII dimer; this complex had the phosphatase activity characteristic of the un-cross-linked PII-NRII complex, and had significant phosphatase activity in the absence of 2 ketoglutarate, suggesting that once PII was tethered to NRII the active conformation was stabilized. Studies with truncated forms of NRII indicated that the purified N-terminal "sensory" domain of NRII was not cross-linked to PII, nor was a polypeptide consisting of NRII residues 1-189. In contrast, polypeptides containing the kinase domain of the transmitter module of NRII (residues 190-349) were cross-linked to PII in an ATP- and 2-ketoglutarate-dependent reaction. These results indicate that PII controls NRII by interaction with the conserved kinase domain of the transmitter module. PMID- 11063582 TI - Sterols and sphingolipids strongly affect the growth of fusion pores induced by the hemagglutinin of influenza virus. AB - Cells expressing the hemagglutinin (HA) of influenza virus were fused to planar phospholipid bilayer membranes to evaluate the effects of sterols and sphingolipids in the target bilayer membranes on properties of fusion pores. Typically, in the absence of sterol, flickering pores are observed, followed by a successful pore (i.e., a pore that fully opens). The incorporation of cholesterol into the lipid bilayer had a marked effect: it greatly decreased the number of flickers, and the first pore formed was usually successful. Similar effects were produced by the sterols epicholesterol and 5beta-cholestanol. In contrast, the sterols cholesteryl acetate, coprostanol, and stanolone did not affect pore flickering, and a successful pore was observed to follow the typical number of flickers. 5alpha-cholestanol gave intermediate results. From these results, it follows that the 3-OH of cholesterol is essential to reduce flickering, but it does not matter if the 3-OH is in an alpha or beta configuration. The double bond is also not critical for the actions of cholesterol nor is the fact that it is a flat molecule. The sphingolipids sphingomyelin, lactosyl cerebroside, and glucosyl cerebroside tended to inhibit full pore enlargement, prolonging the stage of pore flickering. If a sphingolipid and a sterol that strongly interact were both included in the planar membrane, the pattern of flickering was the same as if neither had been included in the bilayer. However, if a sphingolipid and sterol that do not interact with each other were included in the bilayer, the reduced flickering characteristic of the sterol was observed. PMID- 11063583 TI - Mechanism of annexin I-mediated membrane aggregation. AB - It has been proposed that annexin I has two separate interaction sites that are involved in membrane binding and aggregation, respectively. To better understand the mechanism of annexin I-mediated membrane aggregation, we investigated the properties of the inducible secondary interaction site implicated in membrane aggregation. X-ray specular reflectivity measurements showed that the thickness of annexin I layer bound to the phospholipid monolayer was 31 +/- 2 A, indicating that annexin I binds membranes as a protein monomer or monolayer. Surface plasmon resonance measurements of annexin I, V, and mutants, which allowed evaluation of membrane aggregation activity of annexin I separately from its membrane binding, revealed direct correlation between the relative membrane aggregation activity and the relative affinity of the secondary interaction site for the secondary membrane. The secondary binding was driven primarily by hydrophobic interactions, unlike calcium-mediated electrostatic primary membrane binding. Chemical cross linking of membrane-bound annexin I showed that a significant degree of lateral association of annexin I molecules precedes its membrane aggregation. Taken together, these results support a hypothetical model of annexin I-mediated membrane aggregation, in which a laterally aggregated monolayer of membrane-bound annexin I directly interacts with a secondary membrane via its induced hydrophobic interaction site. PMID- 11063584 TI - Coupling of hydrogen bonding to chromophore conformation and function in photoactive yellow protein. AB - To understand in atomic detail how a chromophore and a protein interact to sense light and send a biological signal, we are characterizing photoactive yellow protein (PYP), a water-soluble, 14 kDa blue-light receptor which undergoes a photocycle upon illumination. The active site residues glutamic acid 46, arginine 52, tyrosine 42, and threonine 50 form a hydrogen bond network with the anionic p hydroxycinnamoyl cysteine 69 chromophore in the PYP ground state, suggesting an essential role for these residues for the maintenance of the chromophore's negative charge, the photocycle kinetics, the signaling mechanism, and the protein stability. Here, we describe the role of T50 and Y42 by use of site specific mutants. T50 and Y42 are involved in fine-tuning the chromophore's absorption maximum. The high-resolution X-ray structures show that the hydrogen bonding interactions between the protein and the chromophore are weakened in the mutants, leading to increased electron density on the chromophore's aromatic ring and consequently to a red shift of its absorption maximum from 446 nm to 457 and 458 nm in the mutants T50V and Y42F, respectively. Both mutants have slightly perturbed photocycle kinetics and, similar to the R52A mutant, are bleached more rapidly and recover more slowly than the wild type. The effect of pH on the kinetics is similar to wild-type PYP, suggesting that T50 and Y42 are not directly involved in any protonation or deprotonation events that control the speed of the light cycle. The unfolding energies, 26.8 and 25.1 kJ/mol for T50V and Y42F, respectively, are decreased when compared to that of the wild type (29.7 kJ/mol). In the mutant Y42F, the reduced protein stability gives rise to a second PYP population with an altered chromophore conformation as shown by UV/visible and FT Raman spectroscopy. The second chromophore conformation gives rise to a shoulder at 391 nm in the UV/visible absorption spectrum and indicates that the hydrogen bond between Y42 and the chromophore is crucial for the stabilization of the native chromophore and protein conformation. The two conformations in the Y42F mutant can be interconverted by chaotropic and kosmotropic agents, respectively, according to the Hofmeister series. The FT Raman spectra and the acid titration curves suggest that the 391 nm form of the chromophore is not fully protonated. The fluorescence quantum yield of the mutant Y42F is 1.8% and is increased by an order of magnitude when compared to the wild type. PMID- 11063585 TI - Defining the bilin lyase domain: lessons from the extended phytochrome superfamily. AB - Through pattern searches of genomic databases, new members of the growing family of phytochrome-related genes were identified and used to construct a 130-180 amino acid motif that delimits the bilin lyase domain, a subdomain of the extended phytochrome family that is sufficient for covalent attachment of linear tetrapyrroles (bilins). To test this hypothesis, portions of locus sll0821, a novel phytochrome-related gene from Synechocystis sp. PCC6803 that encodes a large protein with two potential bilin binding sites, were amplified, and the recombinant apoproteins were tested for bilin binding and phytochrome photoactivity. Our experiments indicated that both sites of this protein, termed Cph2 for cyanobacterial phytochrome 2, possessed bilin lyase activity, revealing two distinct classes of bilin lyase domains--those whose bilin adducts are red, far-red reversible and a second class whose bilin adducts are nonphotochromic. Spectroscopic analysis of photochromic phycocyanobilin and fluorescent phycoerythrobilin adducts of a 24-kDa fragment of Cph2 definitively established that the motif identified by pattern searches represents a bona fide bilin lyase domain. Site-directed mutagenesis of highly conserved charged residues within bilin lyase domains of nearly all members of the extended phytochrome superfamily has identified a glutamate residue critical for bilin binding. PMID- 11063586 TI - Mutagenesis of three conserved Glu residues in a bacterial homologue of the ND1 subunit of complex I affects ubiquinone reduction kinetics but not inhibition by dicyclohexylcarbodiimide. AB - Steady-state kinetics of the H(+)-translocating NADH:ubiquinone reductase (complex I) were analyzed in membrane samples from bovine mitochondria and the soil bacterium Paracoccus denitrificans. In both enzymes the calculated K(m) values, in the membrane lipid phase, for four different ubiquinone analogues were in the millimolar range. Both the structure and size of the hydrophobic side chain of the acceptor affected its affinity for complex I. The ND1 subunit of bovine complex I is a mitochondrially encoded protein that binds the inhibitor dicyclohexylcarbodiimide (DCCD) covalently [Yagi and Hatefi (1988) J. Biol. Chem. 263, 16150-16155]. The NQO8 subunit of P. denitrificans complex I is a homologue of ND1, and within it three conserved Glu residues that could bind DCCD, E158, E212, and E247, were changed to either Asp or Gln and in the case of E212 also to Val. The DCCD sensitivity of the resulting mutants was, however, unaffected by the mutations. On the other hand, the ubiquinone reductase activity of the mutants was altered, and the mutations changed the interactions of complex I with short-chain ubiquinones. The implications of the results for the location of the ubiquinone reduction site in this enzyme are discussed. PMID- 11063587 TI - Kinetics and activation thermodynamics of methane monooxygenase compound Q formation and reaction with substrates. AB - The transient kinetics of formation and decay of the reaction cycle intermediates of the Methylosinus trichosporium OB3b methane monooxygenase (MMO) catalytic cycle are studied as a function of temperature and substrate type and deuteration. Kinetic evidence is presented for the existence of three intermediates termed compounds O, P, and P forming after the addition of O(2) to diferrous MMO hydroxylase (H(r)) and before the formation of the reactive intermediate compound Q. The Arrhenius plots for these reactions are linear and independent of substrate concentration and type, showing that substrate does not participate directly in the oxygen activation phase of the catalytic cycle. Analysis of the transient kinetic data revealed only small changes relative to the weak optical spectrum of H(r) for any of these intermediates. In contrast, large changes in the 430 nm spectral region are associated with the formation of Q. The decay reaction of Q exhibits an apparent first-order concentration dependence for all substrates tested, and the observed rate constant depends on the substrate type. The kinetics of the decay reaction of Q yield a nonlinear Arrhenius plot when methane is the substrate, and the rates in both segments of the plot increase linearly with methane concentration. Together these observations suggest that at least two reactions with a methane concentration dependence, and perhaps two methane molecules, are involved in the decay process. When CD(4) is used as the substrate, a large isotope effect and a linear Arrhenius plot are observed. Analogous plots for all other MMO substrates tested (e.g., ethane) are linear, and no isotope effect for deuterated analogues is observed. This demonstrates that a step other than C-H bond breaking is rate limiting for alternative MMO substrates. A two step Q decay mechanism is proposed that provides an explanation for the lack of an isotope effect for alternative MMO substrates and the fact that rate of oxidation of methane by Q exceeds that of many other hydrocarbons with weaker C-H bonds. PMID- 11063588 TI - Binding of the delta subunit to rod phosphodiesterase catalytic subunits requires methylated, prenylated C-termini of the catalytic subunits. AB - PDE6 (type 6 phosphodiesterase) from rod outer segments consists of two types of catalytic subunits, alpha and beta; two inhibitory gamma subunits; and one or more delta subunits found only on the soluble form of the enzyme. About 70% of the phosphodiesterase activity found in rod outer segments is membrane-bound, and is thought to be anchored to the membrane through C-terminal prenyl groups. The recombinant delta subunit has been shown to solubilize the membrane-bound form of the enzyme. This paper describes the site and mechanism of this interaction in more detail. In isolated rod outer segments, the delta subunit was found exclusively in the soluble fraction, and about 30% of it did not coimmunoprecipitate with the catalytic subunits. The delta subunit that was bound to the catalytic subunits dissociated slowly, with a half-life of about 3.5 h. To determine whether the site of this strong binding was the C-termini of the phosphodiesterase catalytic subunits, peptides corresponding to the C-terminal ends of the alpha and beta subunits were synthesized. Micromolar concentrations of these peptides blocked the phosphodiesterase/delta subunit interaction. Interestingly, this blockade only occurred if the peptides were both prenylated and methylated. These results suggested that a major site of interaction of the delta subunit is the methylated, prenylated C-terminus of the phosphodiesterase catalytic subunits. To determine whether the catalytic subunits of the full length enzyme are methylated in situ when bound to the delta subunit, we labeled rod outer segments with a tritiated methyl donor. Soluble phosphodiesterase from these rod outer segments was more highly methylated (4.5 +/- 0.3-fold) than the membrane-bound phosphodiesterase, suggesting that the delta subunit bound preferentially to the methylated enzyme in the outer segment. Together these results suggest that the delta subunit/phosphodiesterase catalytic subunit interaction may be regulated by the C-terminal methylation of the catalytic subunits. PMID- 11063589 TI - Critical role of a subdomain of the N-terminus of the V1a vasopressin receptor for binding agonists but not antagonists; functional rescue by the oxytocin receptor N-terminus. AB - A fundamental issue in molecular pharmacology is to define how agonist:receptor interaction differs from that of antagonist:receptor. The V(1a) receptor (V(1a)R) is a member of a family of related G-protein-coupled receptors that are activated by the neurohypophysial peptide hormone arginine-vasopressin (AVP). Here we define a short subdomain of the N-terminus of the V(1a)R from Glu(37) to Asn(47) that is an absolute requirement for binding AVP and other agonists. In marked contrast to the situation for agonists, deleting this segment has little or no effect on the binding of either peptide or non-peptide antagonists. In addition, we established that this subdomain was crucial for receptor activation and second messenger generation. The oxytocin receptor (OTR) also binds AVP with high affinity but exhibits a different pharmacological profile to the V(1a)R. Substitution of the N-terminus of the V(1a)R with the corresponding sequence from the OTR generated a chimeric receptor (OTR(N)-V(1a)R). The presence of the OTR N terminus recovered high affinity agonist binding such that the OTR(N)-V(1a)R possessed almost wild-type V(1a)R pharmacology and signaling. Consequently, a domain within the N-terminus is required for agonist binding but it does not provide the molecular discriminator for subtype-selective agonist recognition. Cotransfection and peptide mimetic studies demonstrated that this N-terminal subdomain had to be contiguous with the receptor polypeptide to be functional. This study establishes that a segment of the V(1a)R N-terminus has a pivotal role in the mechanism of agonist binding and provides molecular insight into key differences between the interaction of agonists and antagonists with a peptide receptor family. PMID- 11063590 TI - Identifying the lipid-protein interface and transmembrane structural transitions of the Torpedo Na,K-ATPase using hydrophobic photoreactive probes. AB - To identify regions of the Torpedo Na,K-ATPase alpha-subunit that interact with membrane lipid and to characterize conformationally dependent structural changes in the transmembrane domain, we have proteolytically mapped the sites of photoincorporation of the hydrophobic compounds 3-(trifluoromethyl)-3-(m [(125)I]iodophenyl)diazirine ([(125)I]TID) and the phosphatidylcholine analogue [(125)I]TIDPC/16. The principal sites of [(125)I]TIDPC/16 labeling were identified by amino-terminal sequence analysis of proteolytic fragments of the Na,K-ATPase alpha-subunit and are localized to hydrophobic segments M1, M3, M9, and M10. These membrane-spanning segments have the greatest levels of exposure to the lipid bilayer and constitute the bulk of the lipid-protein interface of the Na,K-ATPase alpha-subunit. The extent of [(125)I]TID and [(125)I]TIDPC/16 photoincorporation into these transmembrane segments was the same in the E(1) and E(2) conformations, indicating that lipid-exposed segments located at the periphery of the transmembrane complex do not undergo large-scale movements during the cation transport cycle. In contrast, for [(125)I]TID but not for [(125)I]TIDPC/16, there was enhanced photoincorporation in the E(2) conformation, and this component of labeling mapped to transmembrane segments M5 and M6. Conformationally sensitive [(125)I]TID photoincorporation into the M5 and M6 segments does not reflect a change in the levels of exposure of these segments to the lipid bilayer as evidenced by the lack of [(125)I]TIDPC/16 labeling of these two segments in either conformation. These results suggest that [(125)I]TID promises to be a useful tool for structural characterization of the cation translocation pathway and for conformationally dependent changes in the pathway. A model of the spatial organization of the transmembrane segments of the Na,K ATPase alpha- and beta-subunits is presented. PMID- 11063591 TI - Structure-function study and anti-HIV activity of synthetic peptide analogues derived from viral chemokine vMIP-II. AB - The viral macrophage inflammatory protein II (vMIP-II) shows a broad spectrum interaction with both CC and CXC chemokine receptors including CCR5 and CXCR4, two principal coreceptors for the cellular entry of human immunodeficiency virus type 1 (HIV-1). Recently, we have shown that a synthetic peptide derived from the N-terminus of vMIP-II, designated as V1, is a potent antagonist of CXCR4 but not CCR5 [Zhou, N., et al. (2000) Biochemistry 39, 3782-3787]. In this study, we synthesized a series of new peptides derived from other regions of vMIP-II and characterized their binding activities with both CXCR4 and CCR5. The results provided further support for the notion that the N-terminus of vMIP-II is the major determinant for CXCR4 recognition and that vMIP-II probably interacts with other chemokine receptors such as CCR5 with different sequence and conformational determinants. To understand the structure-function relationship of V1 peptide, its solution conformation was studied using circular dichroism spectroscopy, which showed a random conformation similar to that of the corresponding N terminus in native vMIP-II. In addition, we synthesized a series of mutant analogues of V1 containing alanine, glycine, or phenylalanine substitution at various positions. Residues Val-1, Arg-7, and Lys-9 of V1 peptide were found to be critical for receptor interaction, because single alanine replacement at these positions dramatically decreased peptide binding to CXCR4. In contrast, alanine or phenylalanine substitution at Cys-11 led to significant enhancement in peptide affinity for CXCR4. Finally, we showed that V1 peptide inhibits HIV-1 replication in CXCR4(+) T-cell lines. These studies provide new insights into the structure function relationship of V1 peptide and demonstrate that this peptide may be a lead for the development of therapeutic agents. PMID- 11063592 TI - Mutations of serine 236-237 and tyrosine 302 residues in the human lipoxin A4 receptor intracellular domains result in sustained signaling. AB - Lipoxin A(4) (LXA(4)) is a potent negative modulator of the inflammatory response. The antiinflammatory activities of LXA(4), such as inhibition of agonist-induced polymorphonuclear cell (PMN) chemotaxis and upregulation of beta 2 integrins, require the expression of a G-protein-coupled, high-affinity LXA(4) receptor (LXA(4)R). We now report that stimulation of PMN with proinflammatory agonist N-formyl peptides (FMLP), calcium ionophore A(23187), or phorbol mirystate acetate (PMA) is followed by marked downregulation of LXA(4) binding (B(max) decrease of approximately 45%) and decreased activation of phospholipases A(2) (PLA(2)) and D (PLD). Elucidation of the mechanisms underlying these effects was addressed by structure-function analyses of the intracellular domains of LXA(4)R. Mutant molecule, S236/S237 --> A/G (LXA(4)R(pk)) and Y302 --> F (LXA(4)R(tk)) were obtained by site-directed mutagenesis to yield receptors lacking the putative targets for serine/threonine kinase- or tyrosine kinase dependent phosphorylation. Expression of wild-type and mutated LXA(4)R sequences in CHO and HL-60 cells was used to examine LXA(4) ligand-receptor interactions and signal transduction events. Results indicated that cells expressing LXA(4)R(pk) or LXA(4)R(tk) displayed sustained activation of PLA(2) and PLD in contrast to the transient ones obtained with LXA(4)R(wt) (peak activation at 2-3 min). Moreover, inhibition of LXA(4)-dependent PLA(2) activity by PMA in LXA(4)R(wt) transfected CHO cells was not observed in cells expressing LXA(4)R(pk). Phosphopeptide immunoblotting revealed that the functional differences between wild-type and mutant LXA(4) receptors are accompanied by distinct changes in the receptor protein phosphorylation pattern. Further characterization of these and related LXA(4)R intracellular domains will help to better understand specific events that regulate the antiinflammatory activities of LXA(4). PMID- 11063593 TI - Evidence for a second nucleotide binding site in rat elongation factor eEF-2 specific for adenylic nucleotides. AB - The rat elongation factor eEF-2 catalyzes the translocation step of protein synthesis. Besides its well-characterized GTP/GDP binding properties, we have previously shown that ATP and ADP bind to eEF-2 [Sontag, B., Reboud, A. M., Divita, G., Di Pietro, A., Guillot, D., and Reboud, J. P. (1993) Biochemistry 32, 1976-1980]. However, whether the adenylic and guanylic nucleotide binding sites were different or not remained unclear. To further characterize these sites, eEF 2 was incubated in the presence of N-methylanthraniloyl (Mant) fluorescent derivatives of GTP, GDP, ATP, and ADP. This led to an increase in the probe fluorescence and to a partial quenching of eEF-2 tryptophans in each case. The Mant-derivatives and the unmodified corresponding nucleotides were shown to bind to eEF-2 with a similar affinity. Competition experiments between Mant-labeled and unmodified nucleotides suggested the presence of two different sites binding either guanylic or adenylic nucleotides. A Forster's transfer between tryptophan residues and the Mant-probe is obtained with both the adenylic and the guanylic Mant-nucleotides, and comparison of the transfer efficiencies confirmed the presence of a second binding site specific for adenylic nucleotides. A sequence alignment of EF-Gs with eEF-2s from different species suggests the presence of potential Walker A and B motifs in an insert of the G-domain of eEF-2s from higher eukaryotes. Our results raise the possibility that a site specific for adenylic nucleotides and located in this insert has appeared in the course of evolution although its physiological function is still unknown. PMID- 11063594 TI - Interaction of CBF alpha/AML/PEBP2 alpha transcription factors with nucleosomes containing promoter sequences requires flexibility in the translational positioning of the histone octamer and exposure of the CBF alpha site. AB - Chromatin remodeling at eukaryotic gene promoter sequences accompanies transcriptional activation. Both molecular events rely on specific protein-DNA interactions that occur within these promoter sequences. Binding of CBFalpha/AML/PEBP2alpha (core binding factor alpha/acute myelogenous leukemia/polyoma enhancer binding protein 2alpha) proteins is a key event in both tissue-specific and developmentally regulated osteocalcin (OC) promoter activity. To address linkage between chromatin organization and transcription factor binding, we reconstituted segments of the rat OC gene proximal promoter into mononucleosomes and studied binding of CBFalpha proteins. We analyzed binding of bacterially produced Cbfalpha2Alpha and Cbfalpha2B, two splice variants of the human CBFalpha2 gene, and determined the effect of heterodimerization with the Cbfbeta subunit on binding activity. Our results indicate that binding of the truncated Cbfalpha2A protein to naked DNA is independent of Cbfbeta whereas Cbfalpha2A binding to nucleosomal DNA was enhanced by Cbfbeta. In contrast, the Cbfalpha2B interaction with either naked or nucleosomal DNA was strongly dependent on heterodimerization with the Cbfbeta subunit. Additionally, our results demonstrate that both Cbfalpha2A alone and Cbfalpha2B complexed with Cbfbeta can interact with nucleosomal DNA only if there is a degree of flexibility in the positioning of the histone octamer on the DNA fragment and exposure of the CBFalpha site. This situation was achieved with a DNA segment of 182 bp from the rat OC promoter that preferentially positions mononucleosomes upstream of the CBFalpha binding site and leaves this element partially exposed. Taken together, these results suggest that nucleosomal translational positioning is a major determinant of the binding of CBFalpha factors to nucleosomal DNA. PMID- 11063595 TI - Expression and structural characterization of the recombinant human doppel protein. AB - The doppel protein (Dpl) is a newly recognized prion protein (PrP)-like molecule encoded by a novel gene locus, prnd, located on the same chromosome as the PrP gene. To study the structural features of Dpl, we have expressed recombinant human Dpl corresponding to the putative mature protein domain (residues 24-152) in Escherichia coli. The primary structure of the recombinant Dpl 24-152 was characterized using gel electrophoresis, N-terminal Edman sequencing, matrix assisted laser desorption ionization mass spectrometry, and electrospray ionization mass spectrometry. Dpl 24-152 was shown to contain two disulfide bonds (Cys94-Cys145 and Cys108-Cys140). The secondary structure of Dpl was analyzed using far-UV circular dichroism spectroscopy. Dpl 24-152 was found to be an alpha helical protein having a high helical content (40%). Dpl 24-152 exhibited characteristics of a thermodynamically stable protein that undergoes reversible and cooperative thermal denaturation. In addition, Dpl was found to be soluble and sensitive to proteinase K digestion. Therefore, Dpl 24-152 possesses biochemical properties similar to those of recombinant PrP. This study provides knowledge about the molecular features of human Dpl that will be useful in further investigation into its normal function and the role it may play in neurodegenerative diseases. PMID- 11063596 TI - pH dependence of formation of a partially unfolded state of a Lys 73 --> His variant of iso-1-cytochrome c: implications for the alkaline conformational transition of cytochrome c. AB - The alkaline conformational transition of a lysine 73 --> histidine variant of iso-1-cytochrome c has been studied. The transition has been monitored at 695 nm, a band sensitive to the presence of the heme-methionine 80 bond, at the heme Soret band which is sensitive to the nature of the heme ligand, and by NMR methods. The guanidine hydrochloride dependence of the alkaline conformational transition has also been monitored. The histidine 73 protein has an unusual biphasic alkaline conformational transition at both 695 nm and the heme Soret band, consistent with a three-state process. The conformational transition is fully reversible. An equilibrium model has been developed to account for this behavior. With this model, it has been possible to obtain the acid constant for the trigger group, pK(H), of the low-pH phase from the equilibrium data. A pK(H) value of 6.6 +/- 0.1 in H(2)O was obtained, consistent with a histidine acting as the trigger group. The NMR data for the low-pH phase of the alkaline conformational transition are consistent with an imidazole ligand replacing Met 80. For the high-pH phase of the biphasic alkaline transition, the NMR data are consistent with lysine 79 being the heme ligand. Guanidine hydrochloride m values of 1.67 +/- 0.08 and 1.1 +/- 0.2 kcal mol(-1) M(-1) were obtained for the low- and high-pH phases of the biphasic alkaline transition of the histidine 73 protein, respectively, consistent with a greater structural disruption for the low-pH phase of the transition. PMID- 11063597 TI - Kinetics and reactivity of the flavin and heme cofactors of cellobiose dehydrogenase from Phanerochaete chrysosporium. AB - The flavin cofactor within cellobiose dehydrogenase (CDH) was found to be responsible for the reduction of all electron acceptors tested. This includes cytochrome c, the reduction of which has been reported to be by the reduced heme of CDH. The heme group was shown to affect the reactivity and activation energy with respect to individual electron acceptors, but the heme group was not involved in the direct transfer of electrons to substrate. A complicated interaction was found to exist between the flavin and heme of cellobiose dehydrogenase. The addition of electron acceptors was shown to increase the rate of flavin reduction and the electron transfer rate between the flavin and heme. All electron acceptors tested appeared to be reduced by the flavin domain. The addition of ferric iron eliminated the flavin radical present in reduced CDH, as detected by low temperature ESR spectroscopy, while it increased the flavin radical ESR signal in the independent flavin domain, more commonly referred to as cellobiose:quinone oxidoreductase (CBQR). Conversely, no radical was detected with either CDH or CBQR upon the addition of methyl-1,4-benzoquinone. Similar reaction rates and activation energies were determined for methyl-1,4 benzoquinone with both CDH and CBQR, whereas the rate of iron reduction by CDH was five times higher than by CBQR, and its activation energy was 38 kJ/mol lower than that of CBQR. Oxygen, which may be reduced by either one or two electrons, was found to behave like a two-electron acceptor. Superoxide production was found only upon the inclusion of iron. Additionally, information is presented indicating that the site of substrate reduction may be in the cleft between the flavin and heme domains. PMID- 11063598 TI - Structural preordering in the N-terminal region of ribosomal protein S4 revealed by heteronuclear NMR spectroscopy. AB - Protein S4, a component of the 30S subunit of the prokaryotic ribosome, is one of the first proteins to interact with rRNA in the process of ribosome assembly and is known to be involved in the regulation of this process. While the structure of the C-terminal 158 residues of Bacillus stearothermophilus S4 has been solved by both X-ray crystallography and NMR, that of the N-terminal 41 residues is unknown. Evidence suggests that the N-terminus is necessary both for the assembly of functional ribosomes and for full binding to 16S RNA, and so we present NMR data collected on the full-length protein (200 aa). Our data indicate that the addition of the N-terminal residues does not significantly change the structure of the C-terminal 158 residues. The data further indicate that the N-terminus is highly flexible in solution, without discernible secondary structure. Nevertheless, structure calculations based on nuclear Overhauser effect spectroscopic data combined with (15)N relaxation data revealed that two short segments in the N-terminus, S(12)RRL(15) and P(30)YPP(33), adopt transiently ordered states in solution. The major conformation of S(12)RRL(15) appears to orient the arginine side chains outward toward the solvent in a parallel fashion, while that of P(30)YPP(33) forms a nascent turn of a polyproline II helix. These segments contain residues that are highly conserved across many prokaryotic species, and thus they are reasonable candidates respectively for sites of interaction with RNA and other ribosomal proteins within the intact ribosome. PMID- 11063599 TI - Inhibitors of abscisic acid 8'-hydroxylase. AB - Structural analogues of the phytohormone (+)-abscisic acid (ABA) have been synthesized and tested as inhibitors of the catabolic enzyme (+)-ABA 8' hydroxylase. Assays employed microsomes from suspension-cultured corn cells. Four of the analogues [(+)-8'-acetylene-ABA, (+)-9'-propargyl-ABA, (-)-9'-propargyl ABA, and (+)-9'-allyl-ABA] proved to be suicide substrates of ABA 8'-hydroxylase. For each suicide substrate, inactivation required NADPH, increased with time, and was blocked by addition of the natural substrate, (+)-ABA. The most effective suicide substrate was (+)-9'-propargyl-ABA (K(I) = 0.27 microM). Several analogues were competitive inhibitors of ABA 8'-hydroxylase, of which the most effective was (+)-8'-propargyl-ABA (K(i) = 1.1 microM). Enzymes in the microsomal extracts also hydroxylated (-)-ABA at the 7'-position at a low rate. This activity was not inhibited by the suicide substrates, showing that the 7' hydroxylation of (-)-ABA was catalyzed by a different enzyme from that which catalyzed 8'-hydroxylation of (+)-ABA. Based on the results described, a simple model for the positioning of substrates in the active site of ABA 8'-hydroxylase is proposed. In a representative physiological assay, inhibition of Arabidopsis thaliana seed germination, (+)-9'-propargyl-ABA and (+)-8'-acetylene-ABA exhibited substantially stronger hormonal activity than (+)-ABA itself. PMID- 11063600 TI - Discovery of potent cyclic pseudopeptide human tachykinin NK-2 receptor antagonists. PMID- 11063601 TI - (-)-Spiro[1-azabicyclo[2.2.2]octane-3,5'-oxazolidin-2'-one], a conformationally restricted analogue of acetylcholine, is a highly selective full agonist at the alpha 7 nicotinic acetylcholine receptor. AB - Neuronal nicotinic acetylcholine receptors are members of the ligand-gated ion channel receptor superfamily and may play important roles in modulating neurotransmission, cognition, sensory gating, and anxiety. Because of its distribution and abundance in the CNS, the alpha 7 nicotinic receptor is a strong candidate to be involved in some of these functions. In this paper we describe the synthesis and in vitro profile of AR-R17779, (-)-spiro[1-azabicyclo[2.2. 2]octane-3,5'-oxazolidin-2'-one] (4a), a potent full agonist at the rat alpha 7 nicotinic receptor, which is highly selective for the rat alpha 7 nicotinic receptor over the alpha 4 beta 2 subtype. Preliminary SAR of AR-R17779 presented here indicate that there is little scope for modification of this rigid molecule as even minor changes result in significant loss of the alpha 7 nicotinic receptor affinity. PMID- 11063602 TI - Synthesis of phenalene and acenaphthene derivatives as new conformationally restricted ligands for melatonin receptors. AB - Conformationally restricted phenalene and acenaphthene derivatives 5 were synthesized from phenalen-1-one and acenaphthen-1-one derivatives using the Horner-Emmons reaction. The amines were prepared through the corresponding isocyanates by the Curtius reaction on the acids or by the reduction of the nitriles. Amido derivatives (R(3) = Me, Et, n-Pr, c-Pr) were prepared by acylation of the amines with the appropriate anhydrides or acid chlorides or by the reductive acylation of the nitriles. The affinities of the compounds for melatonin binding sites were evaluated in vitro in binding assays using chicken brain melatonin and the human mt(1) and MT(2) receptors expressed in HEK-293 cells. The functionality of the compounds was determined by the potency to lighten the skin of Xenopus laevis tadpoles. Highly potent compounds were obtained. The data highlighted the role of the methoxy group located in the ortho position to the ethylamido chain as compounds with picomolar affinities such as 14c were obtained (chicken brain, hmt(1), hMT(2) K(i) values = 0.02, 0.008, 0.069 nM, respectively). Compound 14c was equipotent to the corresponding dimethoxy derivative 15c (chicken brain, hmt(1), hMT(2) K(i) values = 0.07, 0.016, 0.1 nM, respectively). On the other hand, the restricted conformation of the amido chain did not influence selectivity for the cloned hmt(1) and hMT(2) receptors. These compounds were also potent agonists of melanophore aggregation in X. laevis. 15a,c were several hundred fold more potent than melatonin (EC(50) = 0.025, 0.004 nM, respectively). Conformational studies indicated that the minimum energy folded conformation of the ethylamido chain could constitute the putative active form in the receptor site in agreement with previous results. PMID- 11063603 TI - Rational design, synthesis, and biological activity of benzoxazinones as novel factor Xa inhibitors. AB - Inappropriate thrombus formation within blood vessels is the leading cause of mortality in the industrialized world. Factor Xa (FXa) is a trypsin-like serine protease that plays a key role in the blood coagulation cascade and represents an attractive target for anticoagulant drug development. From a high-throughput in vitro mass screen of our chemical library, we identified 4-[5-[(2R,6S)-2, 6 dimethyltetrahydro-1(2H)-pyridinyl]pentyl]-2-phenyl-2H-1, 4-benzoxazin-3(4H)-one (1a) as an inhibitor of FXa with an IC(50) of 27 microM. Through a combination of SAR studies and molecular modeling, we synthesized 3-(4-[5-[(2R,6S)-2, 6 dimethyltetrahydro-1(2H)-pyridinyl]pentyl]-3-oxo-3,4-dihydro-2H- 1,4-benzoxazin-2 yl)-1-benzenecarboximidamide (1n) which was a potent FXa inhibitor with an IC(50) of 3 nM. This compound exhibited high selectivity for FXa over other related serine proteases and was efficacious when dosed intravenously in rabbit and dog antithrombotic models. PMID- 11063604 TI - Resistance-modifying agents. 8. Inhibition of O(6)-alkylguanine-DNA alkyltransferase by O(6)-alkenyl-, O(6)-cycloalkenyl-, and O(6)-(2 oxoalkyl)guanines and potentiation of temozolomide cytotoxicity in vitro by O(6) (1-cyclopentenylmethyl)guanine. AB - A series of O(6)-allyl- and O(6)-(2-oxoalkyl)guanines were synthesized and evaluated, in comparison with the corresponding O(6)-alkylguanines, as potential inhibitors of the DNA-repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT). Simple O(6)-alkyl- and O(6)-cycloalkylguanines were weak AGT inactivators compared with O(6)-allylguanine (IC(50) = 8.5 +/- 0.6 microM) with IC(50) values ranging from 100 to 1000 microM. The introduction of substituents at C-2 of the allyl group of O(6)-allylguanine reduced activity compared with the parent compound, while analogous compounds in the O(6)-(2-oxoalkyl)guanine series exhibited very poor activity (150-1000 microM). O(6)-Cycloalkenylguanines proved to be excellent AGT inactivators, with 1-cyclobutenylmethylguanine (IC(50) = 0.55 +/- 0.02 microM) and 1-cyclopentenylmethylguanine (IC(50) = 0.39 +/- 0.04 microM) exhibiting potency approaching that of the benchmark AGT inhibitor O(6) benzylguanine (IC(50) = 0.18 +/- 0.02 microM). 1-Cyclopentenylmethylguanine also inactivated AGT in intact HT29 human colorectal carcinoma cells (IC(50) = 0.20 +/ 0.07 microM) and potentiated the cytotoxicity of the monomethylating antitumor agent Temozolomide by approximately 3- and 10-fold, respectively, in the HT29 and Colo205 tumor cell lines. The observation that four mutant AGT enzymes resistant to O(6)-benzylguanine also proved strongly cross-resistant to 1 cyclopentenylmethylguanine indicates that the O(6)-substituent of each compound makes similar binding interactions within the active site of AGT. PMID- 11063605 TI - Resistance-modifying agents. 9. Synthesis and biological properties of benzimidazole inhibitors of the DNA repair enzyme poly(ADP-ribose) polymerase. AB - The nuclear enzyme poly(ADP-ribose) polymerase (PARP) facilitates the repair of DNA strand breaks and is implicated in the resistance of cancer cells to certain DNA-damaging agents. Inhibitors of PARP have clinical potential as resistance modifying agents capable of potentiating radiotherapy and the cytotoxicity of some forms of cancer chemotherapy. The preclinical development of 2-aryl-1H benzimidazole-4-carboxamides as resistance-modifying agents in cancer chemotherapy is described. 1H-Benzimidazole-4-carboxamides, particularly 2-aryl derivatives, are identified as a class of potent PARP inhibitors. Derivatives of 2-phenyl-1H-benzimidazole-4-carboxamide (23, K(i) = 15 nM), in which the phenyl ring contains substituents, have been synthesized. Many of these derivatives exhibit K(i) values for PARP inhibition < 10 nM, with 2-(4-hydroxymethylphenyl) 1H-benzimidazole-4-carboxamide (78, K(i) = 1.6 nM) being one of the most potent. Insight into structure-activity relationships (SAR) for 2-aryl-1H-benzimidazole-4 carboxamides has been enhanced by studying the complex formed between 2-(3 methoxyphenyl)-1H-benzimidazole-4-carboxamide (44, K(i) = 6 nM) and the catalytic domain of chicken PARP. Important hydrogen-bonding and hydrophobic interactions with the protein have been identified for this inhibitor. 2-(4-Hydroxyphenyl)-1H benzimidazole-4-carboxamide (45, K(i) = 6 nM) potentiates the cytotoxicity of both temozolomide and topotecan against A2780 cells in vitro (by 2.8- and 2.9 fold, respectively). PMID- 11063606 TI - 3D-QSAR CoMFA on cyclin-dependent kinase inhibitors. AB - Several series of cyclin-dependent kinase inhibitors previously prepared in our laboratory were compared using 3D-QSAR (CDK1) and docking (CDK2) techniques. Evaluation of our own library of 93 purine derivatives served to establish the model which was validated by evaluation of an external library of 71 compounds. The best predictions were obtained with the CoMFA standard model (q(2) = 0.68, r(2) = 0.90) and with the CoMSIA combined steric, electrostatic, and lipophilic fields (q(2) = 0.74, r(2) = 0.90). The CDK1 3D-QSAR model was then superimposed to the ATP/CDK2 binding site, giving direct contour maps of the different fields. Although too few compounds were evaluated on CDK5 to derive a 3D-QSAR model, some interesting SARs have been deduced. Comparison of the results obtained from both methods helped with understanding the specific activity of some compounds and designing new specific CDK inhibitors. PMID- 11063607 TI - Active site binding modes of HIV-1 integrase inhibitors. AB - Using the crystal structure of the first complex of the HIV-1 integrase catalytic core domain with an inhibitor bound to the active site, structural models for the interaction of various inhibitors with integrase were generated by computational docking. For the compound of the crystallographic study, binding modes unaffected by crystal packing have recently been proposed. Although a large search region was used for the docking simulations, the ligands investigated here are found to bind preferably in similar ways close to the active site. The binding site is formed by residues 64-67, 116, 148, 151-152, 155-156, and 159, as well as by residue 92 in case of the largest ligand of the series. The coherent picture of possible interactions of small-molecule inhibitors at the active site provides an improved basis for structure-based ligand design. The recurring motif of tight interaction with the two lysine residues 156 and 159 is suggested to be of prime importance. PMID- 11063608 TI - Conformationally constrained anesthetic steroids that modulate GABA(A) receptors. AB - Various cyclic ether and other 3 alpha-hydroxyandrostane derivatives bearing a conformationally constrained hydrogen-bonding moiety were prepared. Their anesthetic potency and their binding affinity for GABA(A) receptors, measured by intravenous administration to mice and inhibition of [(35)S]TBPS binding to rat whole brain membranes, were compared with that of known anesthetic 3 alpha hydroxypregnan-20-ones. Synthetic steroids with similar in vitro and in vivo activities to the endogenous 3 alpha-hydroxypregnan-20-ones all had an ether oxygen on the beta-face of the steroid D-ring. These results suggest that for optimal GABA(A) receptor modulation, the hydrogen bond-accepting substituent should be near perpendicular to the plane of the D-ring on the beta-face of the steroid. PMID- 11063609 TI - Thio- and oxoflavopiridols, cyclin-dependent kinase 1-selective inhibitors: synthesis and biological effects. AB - Flavopiridol analogues, thio- and oxoflavopiridols which contain a sulfur (16) or oxygen (18) atom linker between a chromone ring and the hydrophobic side chain, are selective cyclin-dependent kinase 1 (CDK1) inhibitors with an IC(50) of 110 and 130 nM. These analogues were prepared from key intermediate 7 by substituting the ethyl sulfoxide. Enantio pure intermediate piperidone 10 was obtained from the racemic piperidone 8 via a very efficient "dynamic kinetic resolution" in 76% yield. Hydrophobic side chains such as chlorophenyl or tert-butyl produced potent CDK1 inhibitory activity, while hydrophilic side chains such as pyrimidine or aniline caused a severe reduction in CDK inhibitory activity. These analogues are competitive inhibitors with respect to ATP, and therefore activity was dependent upon the CDK subunit without being affected by the cyclin subunit or protein substrate. Thio- and oxoflavopiridols 16 and 18 are not only selective within the CDK family but also discriminated between unrelated serine/threonine and tyrosine protein kinases. CDK1 selective thio- and oxoflavopiridol analogues inhibit the colony-forming ability of multiple human tumor cell lines and possess a unique antiproliferative profile in comparison to flavopiridol. PMID- 11063610 TI - Adenosine analogues as inhibitors of Trypanosoma brucei phosphoglycerate kinase: elucidation of a novel binding mode for a 2-amino-N(6)-substituted adenosine. AB - As part of a project aimed at structure-based design of adenosine analogues as drugs against African trypanosomiasis, N(6)-, 2-amino-N(6)-, and N(2)-substituted adenosine analogues were synthesized and tested to establish structure-activity relationships for inhibiting Trypanosoma brucei glycosomal phosphoglycerate kinase (PGK), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and glycerol-3 phosphate dehydrogenase (GPDH). Evaluation of X-ray structures of parasite PGK, GAPDH, and GPDH complexed with their adenosyl-bearing substrates led us to generate a series of adenosine analogues which would target all three enzymes simultaneously. There was a modest preference by PGK for N(6)-substituted analogues bearing the 2-amino group. The best compound in this series, 2-amino N(6)- [2''(p-hydroxyphenyl)ethyl]adenosine (46b), displayed a 23-fold improvement over adenosine with an IC(50) of 130 microM. 2-[[2''-(p Hydroxyphenyl)ethyl]amino]adenosine (46c) was a weak inhibitor of T. brucei PGK with an IC(50) of 500 microM. To explore the potential of an additive effect that having the N(6) and N(2) substitutions in one molecule might provide, the best ligands from the two series were incorporated into N(6),N(2)-disubstituted adenosine analogues to yield N(6)-(2''-phenylethyl)-2-[(2'' phenylethyl)amino]adenosine (69) as a 30 microM inhibitor of T. brucei PGK which is 100-fold more potent than the adenosine template. In contrast, these series gave no compounds that inhibited parasitic GAPDH or GPDH more than 10-20% when tested at 1.0 mM. A 3.0 A X-ray structure of a T. brucei PGK/46b complex revealed a binding mode in which the nucleoside analogue was flipped and the ribosyl moiety adopted a syn conformation as compared with the previously determined binding mode of ADP. Molecular docking experiments using QXP and SAS program suites reproduced this "flipped and rotated" binding mode. PMID- 11063611 TI - 2D QSAR modeling and preliminary database searching for dopamine transporter inhibitors using genetic algorithm variable selection of Molconn Z descriptors. AB - In light of the chronic problem of abuse of the controlled substance cocaine, we have investigated novel approaches toward both understanding the activity of inhibitors of the dopamine transporter (DAT) and identifying novel inhibitors that may be of therapeutic potential. Our most recent studies toward these ends have made use of two-dimensional (2D) quantitative structure-activity relationship (QSAR) methods in order to develop predictive models that correlate structural features of DAT ligands to their biological activities. Specifically, we have adapted the method of genetic algorithms-partial least squares (GA-PLS) (Cho et al. J. Comput. -Aided Mol. Des., submitted) to the task of variable selection of the descriptors generated by the software Molconn Z. As the successor to the program Molconn X, which generated 462 descriptors, Molconn Z provides 749 chemical descriptors. By employing genetic algorithms in optimizing the inclusion of predictive descriptors, we have successfully developed a robust model of the DAT affinities of 70 structurally diverse DAT ligands. This model, with an exceptional q(2) value of 0.85, is nearly 25% more accurate in predictive value than a comparable model derived from Molconn X-derived descriptors (q(2) = 0.69). Utilizing activity-shuffling validation methods, we have demonstrated the robustness of both this DAT inhibitor model and our QSAR method. Moreover, we have extended this method to the analysis of dopamine D(1) antagonist affinity and serotonin ligand activity, illustrating the significant improvement in q(2) for a variety of data sets. Finally, we have employed our method in performing a search of the National Cancer Institute database based upon activity predictions from our DAT model. We report the preliminary results of this search, which has yielded five compounds suitable for lead development as novel DAT inhibitors. PMID- 11063612 TI - Sulfoxide-containing aromatic nitrogen mustards as hypoxia-directed bioreductive cytotoxins. AB - A series of diaryl and alkylaryl sulfoxide-containing nitrogen mustards were synthesized and evaluated for their hypoxia-selective cytotoxicity against V-79 cells in vitro as well as for their metabolism profiles with the rat S-9 fractions. In general, the diaryl sulfoxides (4, 5, and 7-9) showed much greater hypoxia selectivity (11-27-fold) than the alkylaryl sulfoxides (approximately 3 fold) (1 and 3). The fused diphenyl sulfoxides (10 and 11), on the other hand, showed very low hypoxia selectivity (1.3-3-fold). Compound 10 was highly cytotoxic under both aerobic and anaerobic conditions, while 11 showed low cytotoxicity under both conditions. The bioreduction of 8 by the rat S-9 fraction under anaerobic conditions was inhibited by menadione and enhanced by benzaldehyde, acetaldehyde, or 2-hydroxypyrimidine suggesting the involvement of aldehyde oxidase in the reduction of the sulfoxides. Bioreductive metabolism studies of selected model sulfoxides suggested that diaryl sulfoxides are better substrates for aldehyde oxidase than alkylaryl sulfoxides. PMID- 11063613 TI - The mitochondrial monoamine oxidase-aldehyde dehydrogenase pathway: a potential site of action of daidzin. AB - Recent studies showed that daidzin suppresses ethanol intake in ethanol preferring laboratory animals. In vitro, it potently and selectively inhibits the mitochondrial aldehyde dehydrogenase (ALDH-2). Further, it inhibits the conversion of monoamines such as serotonin (5-HT) and dopamine (DA) into their respective acid metabolites, 5-hydroxyindole-3-acetic acid (5-HIAA) and 3,4 dihydroxyphenylacetic acid (DOPAC) in isolated hamster or rat liver mitochondria. Studies on the suppression of ethanol intake and inhibition of 5-HIAA (or DOPAC) formation by six structural analogues of daidzin suggested a potential link between these two activities. This, together with the finding that daidzin does not affect the rates of mitochondria-catalyzed oxidative deamination of these monoamines, raised the possibility that the ethanol intake-suppressive (antidipsotropic) action of daidzin is not mediated by the monoamines but rather by their reactive biogenic aldehyde intermediates such as 5-hydroxyindole-3 acetaldehyde (5-HIAL) and/or 3,4-dihydroxyphenylacetaldehyde (DOPAL) which accumulate in the presence of daidzin. To further evaluate this possibility, we synthesized more structural analogues of daidzin and tested and compared their antidipsotropic activities in Syrian golden hamsters with their effects on monoamine metabolism in isolated hamster liver mitochondria using 5-HT as the substrate. Effects of daidzin and its structural analogues on the activities of monoamine oxidase (MAO) and ALDH-2, the key enzymes involved in 5-HT metabolism in the mitochondria, were also examined. Results from these studies reveal a positive correlation between the antidipsotropic activities of these analogues and their abilities to increase 5-HIAL accumulation during 5-HT metabolism in isolated hamster liver mitochondria. Daidzin analogues that potently inhibit ALDH 2 but have no or little effect on MAO are most antidipsotropic, whereas those that also potently inhibit MAO exhibit little, if any, antidipsotropic activity. These results, although inconclusive, are consistent with the hypothesis that daidzin may act via the mitochondrial MAO/ALDH pathway and that a biogenic aldehyde such as 5-HIAL may be important in mediating its antidipsotropic action. PMID- 11063614 TI - Calculation and prediction of binding free energies for the matrix metalloproteinases. AB - The zinc-dependent matrix metalloproteinases are drug targets of interest for diseases ranging from arthritis to cancer. Unfortunately, the use of computational rational drug design has been limited by the challenges introduced by the zinc center. We present an extension of the MM/PB/SA methodology which allows us to calculate the relative binding energies of six known nanomolar carboxylate ligands of MMP-1. We are able to rank the neutral and charged ligands correctly. We further illustrate the utility of our approach by modifying the best-binding ligand of our set and predicting a better binding ligand. PMID- 11063615 TI - Synthesis and biological evaluation of ceramide analogues with substituted aromatic rings or an allylic fluoride in the sphingoid moiety. AB - The biological activity of synthetic ceramide analogues, having modified sphingoid and N-acyl chains, as well as fluorine substituents in the allylic position, was investigated in hippocampal neurons. Their influence on axonal growth was compared to that of C(6)-N-acyl analogues of natural ceramides. D erythro-Ceramides with a phenyl group in the sphingoid moiety and a short N-acyl chain were able to reverse the inhibitory effect of fumonisin B(1) (FB(1)), but not of D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (PDMP), on accelerated axonal growth in hippocampal neurons. Moreover, we demonstrated that a ceramide analogue with an aromatic ring in the sphingoid moiety is recognized as a substrate by glucosylceramide synthase, which suggests that the observed biological effects are mediated by activation of the ceramide analogue via glucosylation. Introduction of a methyl, pentyl, fluoro, or methoxy substituent in the para position of the phenyl ring in the sphingoid moiety yielded partly active compounds. Likewise, substitution of the benzene ring for a thienyl group did not abolish the ability to reverse the inhibition of accelerated axonal growth by FB(1). Both D-erythro- and L-threo-ceramide analogues, having an allylic fluorine substituent, partly reversed the FB(1) inhibition. PMID- 11063616 TI - 3-(3,5-Dimethoxyphenyl)-1,6-naphthyridine-2,7-diamines and related 2-urea derivatives are potent and selective inhibitors of the FGF receptor-1 tyrosine kinase. AB - A series of 3-aryl-1,6-naphthyridine-2,7-diamines and related 2-ureas were prepared and evaluated as inhibitors of the FGF receptor-1 tyrosine kinase. Condensation of 4,6-diaminonicotinaldehyde and substituted phenylacetonitriles gave intermediate naphthyridine-2,7-diamines, and direct reaction of the monoanion of these (NaH/DMF) with alkyl or aryl isocyanates selectively gave the 2-ureas in varying yields (23-93%). For the preparation of more soluble 7 alkylamino-2-ureas, a number of protecting groups for the 2-amine were evaluated (phthaloyl, 4-methoxybenzyl) following selective blocking of the 7-amine (trityl), but these were not superior to the (required) 2-tert-Bu-urea group itself. Direct alkylation of the anion of the (unprotected) 7-amino group with excess 4-(3-chloropropyl)morpholine in DMF gave low (10%) yields of the desired product, but alkylation of the 7-acetamido anion, followed by mild alkaline hydrolysis, raised this to 64%. 3-Phenyl analogues were nonspecific inhibitors of isolated c-Src, FGFR, and PDGFR tyrosine kinases, whereas 3-(2,6-dichlorophenyl) analogues were most effective against c-Src and FGFR, and 3-(3,5-dimethoxyphenyl) derivatives showed high selectivity for FGFR alone. A water-soluble (7 morpholinylpropylamino) analogue retained high FGFR potency (IC(50) 31 nM) and selectivity. Pairwise comparison of the 1, 6-naphthyridines and the corresponding known pyrido[2,3-d]pyrimidine analogues showed little differences in potency or patterns of selectivity, suggesting that the 1-aza atom of the latter is not important for activity. A 7-acetamide derivative inhibited the growth of FGFR expressing tumor cell lines and was particularly potent against HUVECs (IC(50) 4 nM). This compound was also a very potent inhibitor of HUVEC microcapillary formation (IC(50) 0.01 nM) and Matrigel invasion (IC(50) 7 nM) and showed significant in vivo antitumor effects in a highly vascularized mammary adenocarcinoma 16/c model at nontoxic doses. The compounds are worthy of further evaluation as antiangiogenesis agents. PMID- 11063617 TI - Inhibition of protein synthesis by didemnins: cell potency and SAR. AB - Synthetic and naturally occurring didemnins are potent and specific inhibitors of protein synthesis in vitro. Structure-activity analysis indicates a requirement for the intact macrocycle; however, the smaller ring size represented by the didemnin analogue, tamandarin A, is equipotent to didemnin B. Replacement of the N,O-dimethyltyrosine by a N-methylphenylalanine or N-methylleucine residue is also well-tolerated. The rank order for inhibition of protein synthesis in vitro appears to be retained in MCF-7 cells, albeit at much higher potency. This increase in potency is explained for the first time by data indicating that MCF-7 cells can accumulate didemnin B up to 2-3 orders of magnitude compared to the growth medium. PMID- 11063618 TI - Novel bronchodilators: synthesis, transamination reactions, and pharmacology of a series of pyrazino[2,3-c][1,2,6]thiadiazine 2, 2-dioxides. AB - The synthesis, pharmacological evaluation, and structure-activity relationships of a new class of bronchodilator agents, derivatives of pyrazino[2,3 c][1,2,6]thiadiazine 2,2-dioxides are described. The compounds were prepared by reaction of 3,4,5-triamino-1,2, 6-thiadiazine 1,1-dioxide with suitable 1,2 dicarbonyl compounds or alpha-hydroxyiminoketones and subsequent N-alkylation. A transamination procedure for synthesizing derivatives with different substituents at the 4-amino group is reported for the first time. The pyrazino[2,3 c][1,2,6]thiadiazine derivatives were screened for tracheal relaxing activity in vitro, and the active compounds were evaluated in vivo in guinea pigs as bronchodilator agents in comparison to theophylline. Among the compounds studied, the most interesting properties were displayed by the 4-amino-1-ethyl-6-methyl derivative (21). The toxicological evaluation of this derivative is also reported. PMID- 11063619 TI - Syntheses and antimalarial activities of 10-substituted deoxoartemisinins. AB - Two series of 10-substituted deoxoartemisinin derivatives have been synthesized. The first employed the reaction of dihydroartemisinin acetate with several silyl enol ethers in the presence of titanium tetrachloride. The second utilized the reaction of 10-(2-oxoethyl)deoxoartemisinin with several Grignard reagents. The in vitro antimalarial activities of both series were determined against two drug resistant clones of P. falciparum. The activities of 13 beta and 15 beta were 5-7 times greater than that of artemisinin. PMID- 11063620 TI - Synthetic oleanane and ursane triterpenoids with modified rings A and C: a series of highly active inhibitors of nitric oxide production in mouse macrophages. AB - We have designed and synthesized 16 new olean- and urs-1-en-3-one triterpenoids with various modified rings C as potential antiinflammatory and cancer chemopreventive agents and evaluated their inhibitory activities against production of nitric oxide induced by interferon-gamma in mouse macrophages. This investigation revealed that 9(11)-en-12-one and 12-en-11-one functionalities in ring C increase the potency by about 2-10 times compared with the original 12 ene. Subsequently, we have designed and synthesized novel olean- and urs-1-en-3 one derivatives with nitrile and carboxyl groups at C-2 in ring A and with 9(11) en-12-one and 12-en-11-one functionalities in ring C. Among them, we have found that methyl 2-cyano-3, 12-dioxooleana-1,9(11)-dien-28-oate (25), 2-cyano-3,12 dioxooleana-1,9(11)-dien-28-oic acid (CDDO) (26), and methyl 2-carboxy-3,12 dioxooleana-1,9(11)-dien-28-oate (29) have extremely high potency (IC(50) = 0.1 nM level). Their potency is similar to that of dexamethasone although they do not act through the glucocorticoid receptor. Overall, the combination of modified rings A and C increases the potency by about 10 000 times compared with the lead compound, 3-oxooleana-1,12-dien-28-oic acid (8) (IC(50) = 1 microM level). The selected oleanane triterpenoid, CDDO (26), was found to be a potent, multifunctional agent in various in vitro assays and to show antiinflammatory activity against thioglycollate-interferon-gamma-induced mouse peritonitis. PMID- 11063621 TI - Synthesis, biological evaluation, and conformational analysis of A-ring diastereomers of 2-methyl-1,25-dihydroxyvitamin D(3) and their 20-epimers: unique activity profiles depending on the stereochemistry of the A-ring and at C-20. AB - All eight possible A-ring diastereomers of 2-methyl-1, 25-dihydroxyvitamin D(3) (2) and 2-methyl-20-epi-1, 25-dihydroxyvitamin D(3) (3) were convergently synthesized. The A-ring enyne synthons 19 were synthesized starting with methyl (S)-(+)- or (R)-(-)-3-hydroxy-2-methylpropionate (8). This was converted to the alcohol 14 as a 1:1 epimeric mixture in several steps. After having been separated by column chromatography, each isomer led to the requisite A-ring enyne synthons 19 again as 1:1 mixtures at C-1. Coupling of the resulting A-ring enynes 20a-h with the CD-ring portions 5a,b in the presence of a Pd catalyst afforded the 2-methyl analogues 2a-h and 3a-h in good yield. In this way, all possible A ring diastereomers were synthesized. The synthesized analogues were biologically evaluated both in vitro and in vivo. The potency was highly dependent on the stereochemistry of each isomer. In particular, the alpha alpha beta-isomer 2g exhibited 4-fold higher potency than 1 alpha,25-dihydroxyvitamin D(3) (1) both in bovine thymus VDR binding and in elevation of rat serum calcium concentration and was twice as potent as the parent compound in HL-60 cell differentiation. Furthermore, its 20-epimer, that is, 20-epi-alpha alpha beta 3g, exhibited exceptionally high activities: 12-fold higher in VDR binding affinity, 7-fold higher in calcium mobilization, and 590-fold higher in HL-60 cell differentiation, as compared to 1 alpha,25-dihydroxyvitamin D(3) (1). Accordingly, the double modification of 2-methyl substitution and 20 epimerization resulted in unique activity profiles. Conformational analysis of the A-ring by (1)H NMR and an X-ray crystallographic analysis of the alpha alpha beta-isomer 2g are also described. PMID- 11063622 TI - Synthesis and evaluation of novel steroidal oxime inhibitors of P450 17 (17 alpha hydroxylase/C17-20-lyase) and 5 alpha-reductase types 1 and 2. AB - 17 alpha-Hydroxylase/C17-20-lyase (P450 17, CYP 17) and 5 alpha-reductase are the key enzymes in androgen biosynthesis and targets for the treatment of prostate cancer and benign prostatic hyperplasia. In the search of inhibitors for both enzymes, 23 pregnenolone- or progesterone-based steroids were synthesized bearing an oxime group connected directly or via a spacer to the steroidal D-ring. Tested for inhibition of human and rat P450 17, some pregnenolone (9, 11, 14) and a series of progesterone compounds (17-20) turned out to be highly active inhibitors of the human enzyme. The most active compound was Z-21 hydroxyiminopregna-5, 17(20)-dien-3 beta-ol (9) showing K(i) values of 44 and 3.4 nM for the human and rat enzymes, respectively, and a type II UV-difference spectrum indicating a coordinate bond between the oxime group and the heme iron. In contrast to the pregnenolones which showed no inhibition of 5 alpha-reductase isozymes 1 and 2, the progesterones 16, 17, 20, 21, and 23 showed marked inhibition, especially toward the type 2 enzyme. Compounds 17 and 20 were identified as potent dual inhibitors of both P450 17 and 5 alpha-reductase. Tested for selectivity, the most potent P450 17 inhibitors 9, 10, and 14 showed no or only marginal inhibition of P450 arom, P450 scc, and P450 TxA(2). Selected compounds were tested for inhibition of the target enzymes using whole-cell assays. Compounds 9-11 strongly inhibited P450 17 being coexpressed with NADPH P450 reductase in E. coli cells, and 16, 20, and 23 markedly inhibited 5 alpha reductase expressed in HEK 293 cells. Tested for in vivo activity, 9 (0.019 mmol/kg) decreased the plasma testosterone concentration in rats after 2 and 6 h by 57% and 44%. PMID- 11063623 TI - Contribution of the adenine base to the activity of adenophostin A investigated using a base replacement strategy. AB - Syntheses of 3'-O-alpha-D-glucopyranosyl-1-beta-D-ribofuranosidoimidazole 2',3'', 4''-trisphosphate (7) and 3'-O-alpha-D-glucopyranosyl-9-beta-D ribofuranosidopurine 2',3'',4''- trisphosphate (8), two analogues of the superpotent 1D-myo-inositol 1,4,5-trisphosphate receptor agonist adenophostin A (2), are described. 5-O-Benzyl-1, 2-O-isopropylidene-alpha-D-ribofuranose was prepared by an improved route from 1,2-O-isopropylidene-alpha-D-xylofuranose and was coupled with 3,4-di-O-acetyl-2,6-di-O-benzyl-D-glucopyranosyl dimethyl phosphite to give 3',4'-di-O-acetyl-2',5, 6'-tri-O-benzyl-3-O-alpha-D glucopyranosyl-1, 2-O-isopropylidene-alpha-D-ribofuranose. Removal of the isopropylidene acetal and subsequent acetylation gave the central disaccharide 1,2,3',4'-tetra-O-acetyl-2',5, 6'-tri-O-benzyl-3-O-alpha-D-glucopyranosyl-D ribofuranose. Vorbruggen condensation with activated imidazole or purine gave the required beta-substituted derivatives which were further elaborated to 7 and 8, respectively. Radioligand binding assays to hepatic InsP(3) receptors and functional assays of Ca(2+) release from permeabilized hepatocytes gave a rank order of potency of the ligands 2 approximately 8 > 7 approximately Ins(1,4,5)P(3) indicating that the N(6)-amino group of 2 is of little importance for activity and that a minimum of a two-fused-ring nucleobase is required for activity to exceed that of Ins(1,4,5)P(3). The role of the adenine base in the activity of the adenophostins is discussed. This general method should facilitate ready access to nucleobase-modified adenophostin analogues for SAR studies. PMID- 11063624 TI - Structure-activity relationships of a series of pyrrolo[3,2-d]pyrimidine derivatives and related compounds as neuropeptide Y5 receptor antagonists. AB - Neuropeptide Y (NPY) has been shown to play an important role in the regulation of food intake and energy balance. Pharmacological data suggests that the Y5 receptor subtype contributes to the effects of NPY on appetite, and therefore a Y5 antagonist might be a useful therapeutic agent for the treatment of obesity. In attempts to identify potential Y5 antagonists, a series of pyrrolo[3, 2 d]pyrimidine derivatives was prepared and evaluated for their ability to bind to Y5 receptors in vitro. We report here the synthesis and initial structure activity relationship investigations for this class of compounds. The target compounds were prepared by a variety of synthetic routes designed to modify both the substitution and the heterocyclic core of the pyrrolo[3,2-d]pyrimidine lead 1. In addition to identifying several potent Y5 antagonists for evaluation as potential antiobesity agents, a pharmacophore model for the human Y5 receptor is presented. PMID- 11063625 TI - Synthesis and biological activity of novel 5-fluoro-2'-deoxyuridine phosphoramidate prodrugs. AB - A series of novel haloethyl and piperidyl phosphoramidate FdUMP prodrug analogues has been synthesized, and the growth inhibitory activity of these compounds has been evaluated against L1210 mouse leukemia cells. All compounds exhibited potent inhibition of L1210 cell proliferation with IC(50) values in the nanomolar range. Growth inhibition was reversed by the addition of 5 microM thymidine, suggesting a mechanism of action involving the intracellular release of FdUMP. (31)P NMR studies carried out on model haloethyl phosphoramidates confirm the release of nucleotide via cyclization of the phosphoramidate anion to the aziridinium ion intermediate followed by hydrolysis of the P-N bond. The data suggests that <50% of the prodrug is converted to FdUMP intracellularly by this pathway. Piperidyl phosphoramidate analogues are also converted to nucleotide intracellularly, presumably by the action of an endogenous phosphoramidase. PMID- 11063626 TI - Activation mechanisms of nucleoside phosphoramidate prodrugs. AB - A series of thymidine and tetrahydrofurfuryl phosphoramidates bearing haloethyl or piperidyl substituents was synthesized and used to investigate the activation mechanisms of nucleoside phosphoramidate prodrugs. Structure assignments for the tetrahydrofurfuryl reaction products were confirmed by comparison to authentic samples. Structural assignments for thymidine phosphoramidate reaction products were made by analogy to the tetrahydrofurfuryl products. Generation of the phosphoramidate anion leads to cyclization and subsequent nucleophilic attack at carbon and phosphorus of the resulting aziridinium ion intermediate to give the observed products. Nucleophilic attack by water at carbon and phosphorus occurs without selectivity, supporting a mechanism of action of haloethylamine nucleoside prodrugs involving intracellular release of the nucleotide. Activation of the benzotriazolyl piperidyl phosphoramidates is followed by P-N bond hydrolysis; this reaction is subject to specific acid catalysis and to nucleophilic catalysis by 1-hydroxybenzotriazole. These results suggest that the mechanism of action of the piperidyl nucleoside phosphoramidates involves the intracellular release of the active nucleotide following P-N bond cleavage, presumably by the action of an endogenous phosphoramidase. PMID- 11063627 TI - Hydrolytic stability versus ring size in lactams: implications for the development of lactam antibiotics and other serine protease inhibitors. AB - beta-Lactam antibiotics act by acylating a serine hydroxyl group in the catalytic center of bacterial proteases. This requires, among other things, suitable reactivity of the lactam moiety. To evaluate the possible suitability of other lactam systems, kinetic studies were performed using the model reaction of lactams with hydroxide. Following the pace of the reaction by NMR, we found gamma butyrolactam to be hydrolyzed considerably slower than beta-propiolactam. Surprisingly, delta-valerolactam and beta-propiolactam had the same reactivity. beta-Lactam antibiotics were more reactive than both by approximately a factor of 10(3). Medium-sized lactams were least susceptible to hydrolysis. The study highlights the as yet overlooked six-membered lactam ring as a promising vantage point for the development of new classes of antiinfectives and other serine protease inhibitors. PMID- 11063628 TI - Sex: who cares? PMID- 11063629 TI - Recognition and sex categorization of adults' and children's faces: examining performance in the absence of sex-stereotyped cues. AB - The ability of children and adults to classify the sex of children's and adults' faces using only the biologically based internal facial structure was investigated. Face images of 7- to 10-year-old children and of adults in their 20s were edited digitally to eliminate hairstyle and clothing cues to sex. Seven year-olds, nine-year-olds, and adults classified a subset of these faces by sex and were asked, subsequently, to recognize the faces from among the entire set of faces. This recognition task was designed to assess the relationship between categorization and recognition accuracy. Participants categorized the adult faces by sex at levels of accuracy varying from just above chance (7-year-olds) to nearly perfect (adults). All participant groups performed less accurately for children's faces than for adults' faces. The 7-year-olds were unable to classify the children's faces by sex at levels above chance. Finally, the faces of children and adults were equally recognizable--a finding that has theoretical implications for understanding the relationship between categorizing and identifying faces. PMID- 11063630 TI - The role of brothers and sisters in the gender development of preschool children. AB - The study examined whether the sex of older siblings influences the gender role development of younger brothers and sisters of age 3 years. Data on the Pre School Activities Inventory, a measure of gender role behavior that discriminates within as well as between the sexes, were obtained in a general population study for 527 girls and 582 boys with an older sister, 500 girls and 561 boys with an older brother, and 1665 singleton girls and 1707 singleton boys. It was found that boys with older brothers and girls with older sisters were more sex-typed than same-sex singletons who, in turn, were more sex-typed than children with other-sex siblings. Having an older brother was associated with more masculine and less feminine behavior in both boys and girls, whereas boys with older sisters were more feminine but not less masculine and girls with older sisters were less masculine but not more feminine. PMID- 11063631 TI - Gender bias in mothers' expectations about infant crawling. AB - Although boys outshine girls in a range of motor skills, there are no reported gender differences in motor performance during infancy. This study examined gender bias in mothers' expectations about their infants' motor development. Mothers of 11-month-old infants estimated their babies' crawling ability, crawling attempts, and motor decisions in a novel locomotor task-crawling down steep and shallow slopes. Mothers of girls underestimated their performance and mothers of boys overestimated their performance. Mothers' gender bias had no basis in fact. When we tested the infants in the same slope task moments after mothers' provided their ratings, girls and boys showed identical levels of motor performance. PMID- 11063632 TI - Gender biases in children's appraisals of injury risk and other children's risk taking behaviors. AB - Children ages 6, 8, and 10 years were given tasks designed to assess their beliefs about risk of injury from activities. Children were asked to appraise the risk of injury for boys and girls engaging in various play behaviors and to judge the sex of the character in stories about children engaging in activities that result in injuries. Results revealed gender biases in children's appraisals of injury risk: Both boys and girls rated boys as having a lower likelihood of injury than girls even though the boys and girls were engaging in the exact same activities. Children also showed higher accuracy in identifying the sex of the character in stories of boys' injuries than girls' injuries, and accuracy improved with the participant's age. Overall, the results indicate that by the age of 6 years children already have differential beliefs about injury vulnerability for boys and girls. Although boys routinely experience more injuries than girls, children rate girls as having a greater risk of injury than boys. With increasing age, school-age children develop a greater awareness of the ways in which boys and girls differ in risk-taking activities that lead to injury outcomes. PMID- 11063633 TI - Sex differences in spatial cognition, computational fluency, and arithmetical reasoning. AB - Alternative explanations for the male advantage in arithmetical reasoning, as measured by the ability to solve complex word problems, include a male advantage in spatial cognition and a male advantage in computational fluency. The current study was designed to test these competing hypotheses. To this end, 113 male and 123 female undergraduates were administered arithmetical computations and arithmetical reasoning tests, along with an IQ test and a test of spatial cognition. There was no sex difference on the IQ test, but males showed significantly higher mean scores on the arithmetical computations, arithmetical reasoning, and spatial cognition measures. A series of structural equation models indicated that individual differences in arithmetical reasoning were related to individual differences in IQ, spatial abilities, and computational fluency. Moreover, the results suggested that the male advantage in arithmetical reasoning is mediated by the male advantages in both computational fluency and spatial cognition. PMID- 11063634 TI - In search of clinical pathways: illumination or illusion--A cervix cancer survey. PMID- 11063635 TI - Are we there yet? PMID- 11063636 TI - A phase III trial of ifosfamide with or without cisplatin in carcinosarcoma of the uterus: A Gynecologic Oncology Group Study. AB - OBJECTIVE: The aims of this study were to substantiate the previously reported activity of ifosfamide in patients with advanced, persistent, or recurrent carcinosarcoma (mixed mesodermal sarcoma) of the uterus, and to determine whether the addition of cisplatin results in an improved response or survival. Secondarily, we sought to determine the toxicity of ifosfamide-cisplatin in this patient population. METHODS: Patients were randomized to receive ifosfamide (1.5 g/m(2)/day) times 5 days every 3 weeks for eight courses with mesna uroprotection, with or without cisplatin (20 mg/m(2)/day) times 5 days. No patient had received previous chemotherapy. RESULTS: Of 224 patients entered on this study, 30 were ineligible for a variety of reasons, leaving 194 evaluable patients. Early in the study, the dose of the combination regimen was reduced by 20% (1 day) because of toxicity. The investigational arms were balanced for age, grade, and Gynecologic Oncology Group performance status. Percentages of adverse effects reported in 191 patients receiving chemotherapy included (ifosfamide/cisplatin-ifosfamide) grade 3 or 4 granulocytopenia (36/60), grade 3 or 4 anemia (8/17), grade 3 or 4 central nervous system toxicity (19/14), and grade 3 or 4 peripheral neuropathy (1/12). Treatment may have contributed to the deaths of 6 patients treated with full doses of ifosfamide and cisplatin for 5 days. The proportion of patients responding to ifosfamide alone versus ifosfamide cisplatin therapy was (0.36 versus 0.54) overall, 0.47 versus 0.61 for pelvic, 0.21 versus 0.54 for lung, and 0.33 versus 0.40 for "other" metastatic sites of measurable disease. The relative odds ratio of response adjusted for measurable sites of disease was 1.82 (P = 0.03, one-tailed test; 95% lower confidence limit, 1.06). Progression-free survival (PFS) and survival data suggest that the combination offers a slight prolongation of PFS (relative risk, 0.73; 95% upper confidence limit, 0.94; P = 0.02, one-tailed test), but no significant survival benefit (relative risk, 0.80, 95% upper confidence limit, 1.03; P = 0.071, one tailed test). CONCLUSION: The addition of cisplatin to ifosfamide appears to offer a small improvement in progression-free survival over ifosfamide alone in the management of advanced carcinosarcoma of the uterus; the added toxicity may not justify the use of this combination. PMID- 11063637 TI - Identification and preservation of the motoric innervation of the bladder in radical hysterectomy type III. AB - OBJECTIVE: To decrease postoperative morbidity associated with radical hysterectomy Rutledge type III, we identified the parasympathetic innervation of the bladder in the cardinal ligament. METHODS: During laparoscopic dissection of the cardinal ligament, we used 7x magnification on 38 consecutive patients with cervical cancer stages IB1 to IIIA with high risk for parametrial involvement when we performed laparoscopy-assisted radical vaginal hysterectomy type III between August 1997 and January 1999. RESULTS: The middle rectal artery was identified as a landmark separating the vascular from the neural part of the cardinal ligament. The neural part was shown to contain the splanchnic pelvic nerves which anastomose with the pelvic plexus. Following preservation of these neural structures all patients were able to void their bladder spontaneously. Following nerve-sparing technique, patients regained bladder function significantly quicker compared with a control group (n = 28) in which the neural part of the cardinal ligament had not been preserved: suprapubic drainage 11.2 days versus 21.4 days (P = 0.0007). CONCLUSION: Using the middle rectal artery as a landmark the neural part of the cardinal ligament can be preserved, resulting in preservation of the motor function of the bladder. PMID- 11063638 TI - Decreased luteinizing hormone receptor mRNA expression in human ovarian epithelial cancer. AB - OBJECTIVES: The objective of this study was to examine the distribution and cellular localization of luteinizing hormone receptor (LHR) in ovarian epithelial tumors (OETs) and their presumed precursor lesions-ovarian epithelial inclusions (OEIs). The clinicopathologic correlation of the receptor expression in OET was also examined. METHODS: Fifteen microdissected samples of ovarian surface epithelium (OSE), 20 OEIs from benign ovaries, and 141 OETs, including 48 cystadenomas, 33 borderline tumors, 60 carcinomas, and 5 metastatic cancers, were examined for LHR expression by using reverse transcription polymerase chain reaction and in situ hybridization. LHR expression in tumor epithelium and tumor stroma was analyzed separately. The clinicopathologic correlation data were analyzed by standard analysis of variance and contingency table methods. RESULTS: LHR expression was identified in the majority of OSE and OEI samples. In OETs, LHR positivity was found in the epithelial cells in 27% of cases and in the stromal compartment in 37% of cases. LHR-positive stromal cells were mainly luteinized cells. Within the tumor epithelium, LHR expression was detected in 42% of benign, 24% of borderline, and 17% of malignant OETs. LHR expression in tumor stroma showed a similar trend of reduction from benign to malignant OETs. Within the 17 carcinomas, LHR was expressed in the epithelium in 47% of grade 1, 12% of grade 2, and only 5% of grade 3 cancers. The mean age of the LHR-positive group was younger than that of the receptor-negative patients. Compared with mucinous and other types of OETs, serous OETs showed higher LHR expression in the epithelium. Compared with the OETs removed in the different menstrual phases, OETs in the secretory phase showed higher LHR in the tumor stroma than in the proliferative phase. No receptor mRNA was detected in the epithelium of five carcinomas metastatic to the ovary. LHR transcription splicing variants from a single previous report were confirmed in this study. CONCLUSIONS: Malignant OETs have significant reduction of LHR expression compared with precursor lesions and benign and borderline OETs. LHR expression shows a steady decline from low-grade to high-grade ovarian cancer. The presence of LHR receptor in tumor epithelium suggests that luteinizing hormone in serum may have direct influence on tumor growth, whereas the receptor in tumor stroma may be indicative of a paracrine function of LH in the development of OETs. PMID- 11063639 TI - Apoptosis may be an early event of progestin therapy for endometrial hyperplasia. AB - OBJECTIVE: The aim of this study was to investigate the role of apoptosis during progestin therapy for the treatment of endometrial hyperplasia. METHODS: Pre- and posttreatment paraffin-embedded endometrial tissue samples from 19 women with endometrial hyperplasia were examined for changes in glandular cellularity and apoptotic activity related to the administration of progestins. Twelve patients were successfully treated with progestin therapy and 7 patients failed treatment. Glandular cellularity was assessed based on calculating the average number of cells per gland obtained on histologic examination of hematoxylin and eosin stained tissue sections. Apoptotic activity was assessed on the same tissue sections by counting the average number of apoptotic cells per 10 high power fields (hpf) using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay. The effects of progesterone on apoptotic activity in a low-grade endometrial adenocarcinoma cell line (Ishikawa cells) was also examined using an ELISA cell death detection kit. RESULTS: Glandular cellularity significantly decreased with progestin therapy in both treatment outcome groups. The reduction in cells per gland was significantly greater in the group of successfully treated cases compared to the treatment failures (P = 0.005). However, within the successfully treated group, in situ detection of apoptotic cells using the TUNEL assay showed no statistical difference between pre- and posttreatment endometrial samples. Interestingly, a significant decrease in apoptosis was found in posttreatment samples of the group with persistent hyperplasia. The average number of apoptotic cells detected in 10 hpf was reduced from 7.9 prior to treatment to 3.1 after progestin therapy (P = 0.03). In the progesterone-treated Ishikawa cell line, an increase in apoptotic activity started at 24 h, reached a peak at 48 h, and continued up to 72 h of hormone treatment. At 48 h, apoptotic activity was 42.6% greater than in the untreated control (P = 0.04). By 72 h of progesterone treatment, apoptosis was 37.2% greater in the treated cells compared to the noninoculated cells (P = 0.04). CONCLUSIONS: Progestin-induced apoptosis may occur during the early period of treatment for endometrial hyperplasia. Compared to the fully responsive group, persistent endometrial hyperplasia may have intrinsically different molecular mechanisms in response to progestin therapy. PMID- 11063640 TI - Contribution of human papillomavirus testing by hybrid capture in the triage of women with repeated abnormal pap smears before colposcopy referral. AB - OBJECTIVE: The purpose of this work was to evaluate the ability of testing for high-risk human papillomavirus (HPV) types using the hybrid capture technique to predict the presence of cervical intraepithelial neoplasia (CIN) II,III in patients with repeated atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LGSIL) on Pap smears. METHODS: Hybrid capture testing and tissue biopsy were performed on 503 consecutive women with ASCUS or LGSIL on repeated Pap smears who were referred for colposcopy. RESULTS: A highly significant association (P < 0.0001) was found between a positive test for high-risk HPV types and CIN II,III, with an 87.0% positive predictive value and a 95.7% negative predictive value. In 226 women with ASCUS on repeated Pap smears, a positive test for high-risk HPV types had a 85.7% sensitivity and a 97% specificity for CIN II,III. In 277 patients with LGSIL on repeated Pap smears, a positive test for high-risk HPV types had an 88.2% sensitivity and a 94.7% specificity for CIN I,II. Reserving colposcopy examination for women who were positive for high-risk HPV types would have reduced the number of referrals for colposcopy to 24.6% and maintained a sensitivity of 87.0% for CIN II,III. CONCLUSIONS: A positive hybrid capture test for high-risk HPV types was highly sensitive and specific for the presence of CIN II,III in patients with ASCUS and LGSIL on repeated Pap smears. We believe that improved methodology will eventually enable more selective colposcopy referrals without affecting patient safety among these women. PMID- 11063641 TI - Power Doppler angiographic appearance and blood flow velocity waveforms in invasive cervical carcinoma. AB - OBJECTIVES: The aim of this study was to investigate the effectiveness of transvaginal power Doppler angiography in predicting cervical malignancy by detecting intratumoral blood flow and to understand the relationship between squamous cell carcinoma (SCC) serum levels and intratumoral blood flow analysis of invasive cervical carcinoma before treatment. METHODS: Thirty-eight patients with cervical carcinoma (35 with stages Ia to IVb invasive carcinoma, 3 with cervical carcinoma in situ) were enrolled for the evaluation of tumor flow using transvaginal ultrasound accompanied with power Doppler angiography before surgery. The pulsatility, resistance index, and vascular index of tumor flow were measured. Thirty patients with proven healthy cervices were used as the control group. Pretreatment SCC serum levels were obtained in 34 women with cervical carcinoma. RESULTS: The pulsatility index and resistance index were significantly lower in the study group than in the control group (P < 0.0001). The vascular index was also significantly lower in the study group than in the control group (P < 0.0001). There were no significant differences among patients with SCC type and non-SCC type cervical carcinoma (P > 0.05) among the six parameters. There was no significant correlation between the pretreatment SCC serum levels with any of the six parameters obtained from the intratumoral blood flow analysis in the SCC group. CONCLUSIONS: Transvaginal ultrasound with power Doppler angiography is a valuable diagnostic tool for differentiating benign tumors of the cervix from malignant ones. Intratumoral blood flow of the cervix supplied us with practical diagnostic information before surgery and may aid in early prediction and management of cervical carcinoma. The use of transvaginal ultrasound with power Doppler angiography in the grading of vascularity ratio within cervical masses provided more sonographic characteristics among the different subclassifications of cervical cancer and is more useful than color Doppler imaging in the visualization of sonographic morphology. PMID- 11063642 TI - Clear cell carcinoma of the ovary: characterization of its CD44 isoform repertoire. AB - OBJECTIVE: CD44 is a cell surface receptor implicated in tumor metastases. We have previously shown that there is a loss of CD44 splice control in clear cell carcinoma (CCCa) of the ovary. Our aim is to characterize the expression of CD44 3v, -5v, -7v, and -10v in clear cell ovarian tumors and to determine their prognostic value. METHODS: Twenty-two cases of ovarian CCCa were studied for CD44 3v, -5v, -7v, and -10v expression by immunocytochemistry. RESULTS: The primary tumors showed expression of CD44-3v, -5v, -7v, and -10v in 44, 55, 61, and 39% of the cases, respectively. We were able to compare the expression of CD44 in the primary tumor and metastatic sites from the same patient in 7 cases (metastatic sites n = 16). We observed decreased immunoreactivity of CD44-3v, -5v, -7v, and 10v in 67, 100, 93, and 92% of the sites, respectively. CD44-3v and -10v expression was absent in 100% of the nonaffected contralateral ovaries while -7v and -10v were expressed in 1/11 (9%) of them. When CD44-10v was not expressed in the primary tumor, only 18% of the women recurred or died of disease; in contrast, of the cases where it was present, 71% of the women recurred or died of disease (P = 0.049). CONCLUSIONS: There is aberrant alternative mRNA splicing in the development of CCCa of the ovary when compared to the contralateral nonaffected ovary. The expression of CD44-10v correlates with survival. Larger series are needed to further understand the role of CD44 isoforms in ovarian cancer metastases. PMID- 11063643 TI - Chemotherapy for malignant mixed Mullerian tumors of the ovary. AB - OBJECTIVE: The aim of this study was to review the chemotherapy experience at Magee-Womens Hospital for malignant mixed mullerian tumor (MMMT) of the ovary. Patients were treated with either paclitaxel/carboplatin (PC) outpatient chemotherapy or platinum/ifosfamide (PI) inpatient chemotherapy as first- or second-line therapy. METHODS: Thirteen patients diagnosed with MMMT of the ovary after complete surgical staging from 1990 to 1999 were studied retrospectively. Six patients received PC combination chemotherapy, of which 3 patients received PC as first-line treatment. The other 3 patients received PC as second-line therapy. Eight patients were treated with PI. Demographic data, pathology, cytoreductive surgery, treatment, and survival rates were reviewed. Complete clinical response (CR) was defined as the disappearance of all measurable disease or normalization of elevated CA 125 level after chemotherapy. Kaplan-Meier analysis was used for survival analysis. RESULTS: The median survival time of patients receiving PC was 19 months. One patient, after receiving PC as first line treatment, demonstrated a CR and is free of disease beyond 33 months. The median survival time of patients managed with PI was 23 months. Three patients with suboptimal disease demonstrated CR after receiving PI. CONCLUSIONS: Optimal chemotherapy regimen for MMMT of ovary remains to be determined. Platinum-based chemotherapy in combination with ifosfamide or paclitaxel may be active against this rare malignancy. PMID- 11063644 TI - Microsatellite instability is a late event in the carcinogenesis of uterine cervical cancer. AB - OBJECTIVE: The purpose of this study was to elucidate the role of genetic instability and LOH in the progression of cervical cancer and also to analyze for correlations between these genetic abnormalities and the clinicopathological characteristics of cervical cancers. METHODS: Seventy-two DNA samples were obtained from 29 carcinoma in situ, 8 microinvasive carcinoma, and 35 invasive cancers. Seven highly polymorphic microsatellite markers representing the chromosome 3p, 6p, and 6q arms were examined by PCR amplification. RESULTS: Microsatellite instability was detected in 8 of 35 (22.9%) invasive cancers and in 1 of 37 (2.7%) early stage cancers (microinvasive cancer and carcinoma in situ). The incidence of MI was statistically higher in invasive cancers (P < 0.02). On the other hand, loss of heterozygosity (LOH) of chromosome 3p was identified in 6/41 (14.6%) invasive cancers and in 3/27 (11.1%) carcinomas in situ. There was no statistical difference between the two groups. There were no significant correlations between the presence of MI or 3p LOH and clinicopathological characteristics including the histological type, FIGO stage, depth of myometrial invasion, lymphovascular involvement, lymph node metastasis, and recurrence. CONCLUSION: Our results indicate that genomic instability is a late event during the carcinogenesis of cervical cancer and is associated with the conversion of cervical intraepithelial neoplasia to an invasive phenotype. To the contrary, LOH of chromosome 3p plays an early role in the development of cervical intraepithelial neoplasia. No significant correlation was observed between the presence of MI or LOH and clinicopathological characteristics. PMID- 11063645 TI - Adenocarcinoma in situ of the uterine cervix: an experience with 100 cases. AB - OBJECTIVE: The aim of this study was to ascertain whether cold knife conization alone for cervical adenocarcinoma in situ is safe. METHODS: One hundred consecutive patients with a histologically proven adenocarcinoma in situ (AIS) of the cervix were studied from 1970 to 1992. RESULTS: Ninety-two women presented with abnormal smears, and of these 56% contained abnormal glandular cells. Sixty seven (74%) of 90 women who underwent colposcopy had an abnormal examination, but a glandular abnormality was suspected in only 19 (28%). In all, 80 cold knife conizations were performed. In 7, no abnormality was found following punch biopsy. The margins were free of disease in 55 (75%). The most commonly involved margin in the remainder was the apical. Conization was followed by hysterectomy in 20 women: in 8 of these the cone margins were free and residual disease was found in 2 of the extirpated uteri: as these were extramural cases, inadequate sampling could not be excluded. Of the 12 women where hysterectomy followed conization with diseased margins, 9 had residual disease in the hysterectomy specimen. The definitive therapy was cold knife conization in 56 patients, hysterectomy in 38, and electrocoagulation diathermy in 6. Follow-up of the 53 patients treated by conization alone ranging from 1 to 16 years, with a mean of 8 years (3 have been lost to follow-up) revealed no recurrence of AIS or adenocarcinoma to date. CONCLUSION: It is concluded that cold knife conization is a safe therapeutic modality, provided that the cone biopsy has been adequately sampled and the margins are free. PMID- 11063646 TI - Treatment of relapsed carcinoma of the ovary with single-agent paclitaxel following exposure to paclitaxel and platinum employed as initial therapy. AB - OBJECTIVES: The aim of this study was to evaluate the ability of paclitaxel to achieve a second clinical response in patients with recurrent epithelial ovarian carcinoma who responded to standard therapy with platinum and paclitaxel in the initial setting. METHODS: Thirty-four patients with epithelial ovarian who demonstrated a complete response to paclitaxel and platinum in the initial treatment setting were retreated with paclitaxel as a single agent for relapse of their disease. Paclitaxel was given at a dose of 135-175 mg/m(2) over 3 h at 21 day intervals. Fifteen patients had platinum-resistant disease and 19 had potentially platinum-sensitive disease. Response was documented by physical examination, serial serum CA125 measurement, or radiologic evaluation. RESULTS: An objective response to paclitaxel retreatment was demonstrated in 15 patients (44%), with a median progression-free interval (PFI) of 8.6 months (range 4-17 months). An additional 14 patients (41%) demonstrated disease stabilization, with a median PFI of 7.4 months (range 3-13 months). Overall, retreatment with paclitaxel was well tolerated, with minimal cumulative toxicities, despite repetitive dosing. CONCLUSION: These results demonstrate that patients with ovarian cancer who relapse after initial treatment with paclitaxel often have disease that is still responsive to the agent. Given its relative lack of cumulative toxicity, retreatment with paclitaxel as a single agent is a reasonable therapeutic option for patients with recurrent ovarian cancer. PMID- 11063647 TI - Glycodelin-A expression in the uterine cervix. AB - OBJECTIVE: Local immunosuppressive factors in the uterine cervix infected by human papillomavirus are felt to facilitate the malignant transformation process. Glycodelin-A is an immunosuppressive peptide found in several tissues of mullerian origin, most notably the pregnant and decidualized endometrium. Its expression in the uterine cervix has not been defined but could theoretically contribute to the immunopermissive environment of the cervix. To determine whether glycodelin-A is found in the cervix we examined the squamous and endocervical epithelia from both normal and neoplastic cervical specimens from 14 women. METHODS: Immunohistochemisty identification of glycodelin-A was performed on archival paraffin-embedded sections from 10 hysterectomies and 4 cone biopsies. Sections were evaluated and staining was scored as negative, positive, or strongly positive with a separate score for the squamous and glandular components of the cervix. RESULTS: Eleven of 14 cases, 79%, demonstrated positive staining of the squamous epithelium. Glycodelin-positive cases included hisologically normal (n = 4; 3 strongly positive, 1 positive) as well as dysplastic (n = 5; 1 strongly positive, 2 positive, and 2 negative) and malignant squamous cells (n = 5; 1 strongly positive, 3 positive, and 1 negative). Normal glandular epithelia were negative in all cases but 1, which demonstrated significant squamous and tubal metaplasia of the endocervical glands involved. CONCLUSION: Glycodelin-A is found in the squamous epithelium of both the histologically normal and the neoplastic cervix. Further characterization of these results will focus on the possible immunosuppressive effect glycodelin-A may have in the cervix. PMID- 11063648 TI - HPV subtype analysis in lower genital tract neoplasms of female renal transplant recipients. AB - OBJECTIVE: Immunocompromised patients, such as female renal transplant recipients, have an increased incidence of neoplasms involving the lower genital tract (i.e., cervix, vagina, vulva). The relationship between lower genital tract neoplasms and human papillomavirus (HPV) infection has been established and high risk oncogenic subtypes have been identified (HPV 16, 18, 45, and 56). The purpose of this study is to evaluate HPV subtypes present in lower genital tract neoplasms of post renal transplant women and compare HPV subtypes found in these patients with immunocompetent patients having similar neoplasms and normal immunocompetent controls. METHODS: Twenty specimens from lower genital tract neoplasms of 16 renal transplant patients, 13 specimens from 13 immunocompetent patients with similar histology, and 13 patients with normal lower genital tract histology were analyzed for the presence of HPV using polymerase chain reaction. HPV primers including the L1 (late) region consensus primers and primers specific for the HPV E6 (early) region for subtypes 6, 11, 16, and 18 were amplified with DNA from the above patient samples. RESULTS: Overall, HPV was detected in 21/46 specimens tested. Thirteen of the HPV-positive specimens were from transplant patients, and 8 were from immunocompetent patients (5 immunocompetent with disease and 3 normal patients). This difference in the total number of HPV positive cases was statistically significant between the transplant and immunocompetent group (P = 0.02). Although no difference in HPV 6 and/or 11 was detected between the two groups, HPV subtypes 16 and/or 18 approached statistical significant difference (P = 0.06). CONCLUSIONS: High-risk oncogenic HPV subtypes 16 and/or 18 were found at a higher rate in transplant patients compared with their immunocompetent counterparts. The combination of immunocompromise and increased HPV 16 and/or 18 positivity may place these patients at increased risk for aggressive lower genital tract neoplastic progression. PMID- 11063649 TI - Proliferation in "atypical" atrophic pap smears. AB - OBJECTIVE: Atrophic cervical epithelium of postmenopausal women may mimic high grade cervical intraepithelial neoplasia (CIN2-3) in Papanicolaou-stained cervical smears (Pap smears). Women with such an "atypical" Pap smear need a repeated Pap smear after a course of estrogens before a definite diagnosis can be made. The aim of this study was to determine whether measurement of proliferative activity in Pap smears of postmenopausal patients that were difficult to interpret is a reliable test for differentiating between cervical atrophy and high-grade CIN. METHODS: Pap smears obtained before and after estrogen treatment of 30 postmenopausal women with an atypical Pap smear were restained with the monoclonal antibody MIB1 to visualize proliferating cells. The proliferative activity index (PAI) was subsequently measured in order to explore the feasibility of a recently proposed PAI-based diagnostic decision tree to reduce the number of estrogen courses and follow-up Pap smears in postmenopausal women. RESULTS: The PAI-based test to discriminate between cervical atrophy and high grade CIN resulted in 100 and 96% correct diagnoses in women with high-grade CIN and cervical atrophy, respectively. Only 2 of the 30 women would have needed a repeated Pap smear after estrogen treatment for definite diagnosis if the PAI based diagnostic decision had been used. CONCLUSIONS: Measurement of PAI in MIB1 restained Pap smears is a simple, reliable, safe, and probably also cost effective method to obtain a substantial reduction of diagnostic estrogen courses and subsequent Pap smears in postmenopausal women with an atypical Pap smear. PMID- 11063650 TI - Inverse expression of Cdk4 and p16 in epithelial ovarian tumors. AB - OBJECTIVES: The aim of this study was to elucidate the expression and the correlation of two cell cycle regulators, cdk4 and its inhibitor p16, in a series of benign, borderline, and malignant ovarian tumors and to evaluate whether their alterations correlate with clinicopathologial parameters and patients' prognosis in epithelial ovarian carcinomas. METHODS: Immunohistochemical analysis using anti-cdk4 and anti-p16 antibodies was carried out for 103 paraffin sections of ovarian tumors, and Western blot analysis and cdk4 activity assay were performed in 26 fresh ovarian tumor samples. RESULTS: The results of immunohistochemistry showed that 60.61 and 69.70% of benign, 69.57 and 56.52% of borderline, and 74.47 and 40. 43% of malignant tumors expressed cdk4 and p16, respectively, demonstrating increased cdk4 and decreased p16 expression in ovarian carcinomas. A significant inverse relationship between cdk4 and p16 expression was found. The loss of p16 expression was more correlated with G(2) and G(3) tumors in contrast with G(1) tumors. No significant correlation was observed between cdk4 expression and clinicopathological parameters. Neither cdk4 nor p16 expression has significant effects on overall survival by the Kaplan-Meier method. When the combined phenotypes of the two proteins were analyzed, patients with cdk4 positive/p16-negative expression had a reduced overall survival than other phenotypes of cdk4/p16. CONCLUSIONS: These data suggest that inverse expression of cdk4 and p16 may be involved in the development and progression of epithelial ovarian carcinomas. The combined phenotypes of cdk4 and p16 proteins could provide a useful prognostic indicator for patients with epithelial ovarian carcinoma. PMID- 11063651 TI - Intraoperative lymphatic mapping in cervix cancer patients undergoing radical hysterectomy: A pilot study. AB - OBJECTIVE: Intraoperative lymphatic mapping and sentinel lymph node identification (SLN) have been increasingly evaluated in the treatment of a variety of solid tumors, particularly breast cancer and melanoma. We sought to evaluate the feasibility of these procedures in patients undergoing radical hysterectomy with pelvic lymphadenectomy for treatment of early cervical cancer. METHODS: Twenty patients with normal-appearing lymph nodes underwent intracervical injection of isosulfan blue dye (lymphazurin 1%) at the time of planned radical hysterectomy and bilateral pelvic/low paraortic lymphadenectomy (40 nodal basins). Regional lymphatic tissue was inspected for dye uptake into lymphatic channels and lymph nodes. Tumor characteristics, surgical findings, and specific locations of lymphatic dye uptake were recorded and correlated with final pathology results. RESULTS: Sentinel lymph nodes were identified in 12 of 20 (60%) patients. A total of 23 sentinel nodes were identified in 17 of 40 (43%) nodal basins dissected (range: 0-2 per basin). Successful SLN identification was less likely in patients with tumors >4 cm compared with those with tumors ==50 years and parity >==5 were significant factors associated with residual disease. The incidence of residual disease was 56.5 and 29. 3% in patients >==50 and <50 years, respectively, and 61.8 and 36.0% in patients with parity >==5 and <5. Post-cone endocervical curettage (ECC) and multiple-quadrant disease were the only pathologic predictive factors identified. The incidence of residual disease was 64.6 and 29.2% in patients with positive ECC and negative ECC, respectively, and 48.4 and 25.9% in patient with multiple-quadrant disease and one- or two quadrant disease. Other pathologic parameters, including endocervical margins, ectocervical margins, endocervical gland involvement, and depth of conization, were not predictive of residual disease. When ECC was combined individually with age, endocervical margins, or multiple-quadrant disease, there was no increase of positive predictive rate. CONCLUSIONS: (1) Age 50 years or more and parity >==5 were two demographic features that predicted post-cone residual disease. (2) ECC and multiple-quadrant disease were the only pathologic parameters that predicted post-cone residual disease. (3) With the appropriate application of the predictive factors, post-cone hysterectomy may be further decreased. PMID- 11063659 TI - A comparison of prognoses of pathologic stage Ib adenocarcinoma and squamous cell carcinoma of the uterine cervix. AB - OBJECTIVES: The influence of the histology of adenocarcinoma on recurrence and survival for patients treated with radical hysterectomy and diagnosed as having pathologic stage Ib cervical cancer was investigated. METHODS: Five hundred and nine patients (405 squamous cell carcinomas, 104 adenocarcinomas) with pathologic stage Ib cervical cancer treated initially at the Aichi Cancer Center between 1976 and 1995 were studied. RESULTS: Multivariate analysis identified the prognostic variables as histology of adenocarcinoma, number of lymph nodes involved, and tumor size beyond 4 cm. Five-year overall survival and disease-free survival of patients with adenocarcinoma in the presence of lymph node metastasis were 63.2 and 47.4%, respectively, significantly poorer than for squamous cell carcinoma (83.6 and 80.6%; P < 0.001 and P = 0.002, respectively). These were not different in the absence of lymph node metastasis (adenocarcinoma, 93.9 and 92.7%; squamous cell carcinoma, 97.9% and 96.1%; P = 0.067 and P = 0.250, respectively). CONCLUSIONS: The independent significant risk factors for the recurrence and survival of pathologic stage Ib cervical cancer were the presence of lymph node metastasis, large tumor size beyond 4 cm, and histology of adenocarcinoma. The prognosis of patients with adenocarcinoma was poorer than of patients with squamous cell carcinoma in the presence of lymph node metastasis, while the prognosis of pathologic stage Ib cervical cancer was equivalent when there was no metastasis. PMID- 11063660 TI - Persistence of human papillomavirus infection after therapeutic conization for CIN 3: is it an alarm for disease recurrence? AB - OBJECTIVE: The aims of this study were (1) to examine whether HPV DNA is persistently detected in the cervix after therapeutic conization for CIN 3 and (2) to explore whether a patient with persistence of HPV infection is at risk of developing recurrent disease. METHODS: Of 74 patients referred with CIN 3, 58 who were tested for HPV DNA in the pretreatment cervical lesions were enrolled in the study. After standard therapeutic conization, patients were followed prospectively at the outpatient clinic. Our follow-up protocol was to follow patients without therapeutic intervention as long as they developed no recurrence or recurrence of CIN 1 or 2, while patients who experienced recurrence of CIN 3 were recommended for reconization or hysterectomy. The polymerase chain reaction for detecting HPV DNA was performed using fresh cell samples from the cervix. RESULTS: In 56 of 58 patients (96.6%), HPV DNAs were detected in their primary cervical lesions prior to conization. With regard to the distribution of HPV types, HPV type 16 family (types 16, 31, and 35) was identified in 28 cases (50.0%), type 18 family (types 18, 33 and 58) in 15 (26.8%), and type X in 18 (32.1%). Up to August 1999, all of the 58 patients have been followed with a mean follow-up period of 31.8 months (range: 12 to 73 months). After treatment, HPV DNA was persistently detected in 11 (19.6%) but negative in 45 (80.4%) of 56 HPV DNA-positive patients. HPV DNA was not detected in both HPV DNA-negative patients. Five of 11 persistently HPV DNA-positive patients (45.5%) developed CIN recurrence, while none of 45 persistently HPV DNA-negative patients did. Thus, there was a significant difference between the recurrence rates of these two groups (P < 0.0001). Both patients who were initially HPV DNA-negative developed no recurrence. Accordingly, the overall recurrence following conservative treatment for CIN 3 was 5 of 58 patients (8.6%). CONCLUSIONS: Patients with persistent HPV infection after conization for CIN 3 should be especially closely followed because they are at increased risk of developing disease recurrence. PMID- 11063661 TI - Assessment of proliferation index with MIB-1 as a prognostic factor in radiation therapy for cervical cancer. AB - OBJECTIVES: The relationship between the expression of murine monoclonal antibody MIB-1, which reacts with Ki-67 nuclear antigen, a marker for proliferating cells, and the prognosis of stage IIIb cervical cancer after radiation therapy was analyzed. METHODS: A total of 67 patients with stage IIIb cervical cancer who had received radiation therapy were included in the retrospective study. The labeled streptavidin-biotin method was used for immunohistochemical staining of the MIB-1 protein. RESULTS: In 32 patients showing a high MIB-1 index (percentage of cells labeled with MIB-1 >/=26.4%), the cumulative 5- and 8-year survival rates were 75.8 and 61.5%, respectively, significantly better (P < 0.05) than those in 35 patients with a low MIB-1 index (<26.4%) (59.6 and 41.1%, respectively). Serum squamous cell carcinoma antigen levels, an index of the response to radiation therapy, decreased to 6-fold in a dosage-dependent manner the total amount of ALDP (mutated and normal) remained approximately even as demonstrated by western blot and flow cytometric analyses. Thus, apparently mutated and normal ALDP compete for integration into a limited number of sites in the peroxisomal membrane. Consequently, increased amounts of mutated ALDP resulted in decreased peroxisomal beta-oxidation and accumulation of very long-chain fatty acids. These findings have direct implications on future gene therapy approaches for treatment of X-ALD, since in some patients a non functional endogenous protein could act in a dominant negative way or displace the introduced, normal protein. PMID- 11063721 TI - A human PKD1 transgene generates functional polycystin-1 in mice and is associated with a cystic phenotype. AB - Three founder transgenic mice were generated with a 108 kb human genomic fragment containing the entire autosomal dominant polycystic kidney disease (ADPKD) gene, PKD1, plus the tuberous sclerosis gene, TSC2. Two lines were established (TPK1 and TPK3) each with approximately 30 copies of the transgene. Both lines produced full-length PKD1 mRNA and polycystin-1 protein that was developmentally regulated, similar to the endogenous pattern, with expression during renal embryogenesis and neonatal life, markedly reduced at the conclusion of renal development. Tuberin expression was limited to the brain. Transgenic animals from both lines (and the TPK2 founder animal) often displayed a renal cystic phenotype, typically consisting of multiple microcysts, mainly of glomerular origin. Hepatic cysts and bile duct proliferation, characteristic of ADPKD, were also seen. All animals with two copies of the transgenic chromosome developed cysts and, in total, 48 of the 100 transgenic animals displayed a cystic phenotype. To test the functionality of the transgene, animals were bred with the Pkd1(del34) knockout mouse. Both transgenic lines rescued the embryonically lethal Pkd1(del34/del34) phenotype, demonstrating that human polycystin-1 can complement for loss of the endogenous protein. The rescued animals were viable into adulthood, although more than half developed hepatic cystic disease in later life, similar to the phenotype of older Pkd1(del34/+) animals. The TPK mice have defined a minimal area that appropriately expresses human PKD1. Furthermore, this model indicates that over-expression of normal PKD1 can elicit a disease phenotype, suggesting that the level of polycystin-1 expression may be relevant in the human disease. PMID- 11063722 TI - Replication protein A1 reduces transcription of the endothelial nitric oxide synthase gene containing a -786T-->C mutation associated with coronary spastic angina. AB - We recently reported that a mutation (-786T-->C) in the promoter region of the endothelial nitric oxide synthase (eNOS) gene reduced transcription of the gene and was strongly associated with coronary spastic angina and myocardial infarction. To elucidate the molecular mechanism for the reduced eNOS gene transcription, we have now purified a protein that specifically binds to the mutant allele in nuclear extracts from HeLa cells. The purified protein was identical to replication protein A1 (RPA1), known as a single-stranded DNA binding protein essential for DNA repair, replication and recombination. In human umbilical vein endothelial cells, inhibition of RPA1 expression using antisense oligonucleotide restored transcription driven by the mutated promoter sequence, whereas, conversely, overexpression of RPA1 further reduced it. RPA1 was similarly detected in placenta and eNOS mRNA levels in placentas carrying the 786T-->C mutation were significantly lower than in placentas without it. The functional importance of the diminished eNOS expression was revealed by the finding that serum nitrite/nitrate levels among individuals carrying the -786T- >C mutation were significantly lower than among those without the mutation. RPA1 thus apparently functions as a repressor protein in the -786T-->C mutation related reduction of eNOS gene transcription associated with the development of coronary artery disease. PMID- 11063723 TI - TRF1 is a critical trans-acting factor required for de novo telomere formation in human cells. AB - The duplex telomere repeat (TTAGGG)(n) is an essential cis-acting element of the mammalian telomere, and an exogenous telomere repeat can induce chromosome breakage and de novo telomere formation at the site of a break (telomere seeding). Telomere seeding requires the telomere repeat (TTAGGG)(n) more stringently than does an in vitro telomerase assay, suggesting that it reflects the activity of a critical trans-acting element of the functional telomere, in addition to telomerase. Furthermore, telomere seeding is induced at a frequency fluctuating widely among human cell lines, suggesting variation in the activity of this hypothetical factor among cells. In this study, we investigated the cellular factor(s) required for telomere formation using the frequency of telomere seeding as an index and identified TRF1, one of the telomere repeat binding proteins, as an essential trans-acting factor. The exogenous telomere repeat induces telomere formation at a frequency determined by the availability of TRF1, even in telomerase-negative cells. Our study shows clearly that TRF1 has a novel physiological significance distinct from its role as a regulator of telomere length in the endogenous chromosome. The possible role of TRF1 in cell aging and immortalization is discussed. PMID- 11063724 TI - Large-scale methylation analysis of human genomic DNA reveals tissue-specific differences between the methylation profiles of genes and pseudogenes. AB - Cytosine in CpG dinucleotides is frequently found to be methylated in the DNA of higher eukaryotes and differential methylation has been proposed to be a key element in the organization of gene expression in man. To address this question systematically, we used bisulfite genomic sequencing to study the methylation patterns of three X-linked genes and one autosomal pseudogene in two adult individuals and across nine different tissues. Two of the genes, SLC6A8 and MSSK1, are tissue-specifically expressed. CDM is expressed ubiquitously. The pseudogene, psi SLC6A8, is exclusively expressed in the testis. The promoter regions of the SLC6A8, MSSK1 and CDM genes were found to be essentially unmethylated in all tissues, regardless of their relative expression level. In contrast, the pseudogene psi SLC6A8 shows high methylation of the CpG islands in all somatic tissues but complete demethylation in testis. Methylation profiles in different tissues are similar in shape but not identical. The data for the two investigated individuals suggest that methylation profiles of individual genes are tissue specific. Taken together, our findings support a model in which the bodies of the genes are predominantly methylated and thus insulated from the interaction with DNA-binding proteins. Only unmethylated promoter regions are accessible for binding and interaction. Based on this model we propose to use DNA methylation studies in conjunction with large-scale sequencing approaches as a tool for the prediction of cis-acting genomic regions, for the identification of cryptic and potentially active CpG islands and for the preliminary distinction of genes and pseudogenes. PMID- 11063725 TI - The Fanconi anemia protein FANCF forms a nuclear complex with FANCA, FANCC and FANCG. AB - Fanconi anemia (FA) is a chromosomal instability syndrome associated with a strong predisposition to cancer, particularly acute myeloid leukemia and squamous cell carcinoma. At the cellular level, FA is characterized by spontaneous chromosomal breakage and a unique hypersensitivity to DNA cross-linking agents. Complementation analysis has indicated that at least seven distinct genes are involved in the pathogenesis of FA. Despite the identification of four of these genes (FANCA, FANCC, FANCF and FANCG), the nature of the 'FA pathway' has remained enigmatic, as the FA proteins lack sequence homologies or motifs that could point to a molecular function. To further define this pathway, we studied the subcellular localizations and mutual interactions of the FA proteins, including the recently identified FANCF protein, in human lymphoblasts. FANCF was found predominantly in the nucleus, where it complexes with FANCA, FANCC and FANCG. These interactions were detected in wild-type and FA-D lymphoblasts, but not in lymphoblasts of other FA complementation groups. This implies that each of the FA proteins, except FANCD, is required for these complexes to form. Similarly, we show that the interaction between FANCA and FANCC is restricted to wild-type and FA-D cells. Furthermore, we document the subcellular localization of FANCA and the FANCA/FANCG complex in all FA complementation groups. Our results, along with published data, culminate in a model in which a multi-protein FA complex serves a nuclear function to maintain genomic integrity. PMID- 11063726 TI - Integrated analysis of sequence evolution and population history using hypervariable compound haplotypes. AB - We have examined compound haplotypes from a highly informative region of human chromosome 16, in which information from the rapid evolution of a highly unstable minisatellite is integrated with data on the longer-term evolution of this segment from 10 flanking substitutional polymorphisms. Combined with sequence data from non-human primates, analysis of relationships between these compound haplotypes allows the reconstruction of a rooted network of the evolutionary pathways between them. Most relationships can be explained via simple substitutional mutations, although the origins of some haplotypes involve recurrent events at a hotspot for substitutional mutation and/or gene conversion. For compound haplotypes including the minisatellite array, the network found in a range of world-wide populations constitutes a highly informative data set for the analysis of population history (437 different compound haplotypes were discriminated among 658 studied). Since the mutation rates and processes of the minisatellite array are known from direct studies, ages for individual lineages have been estimated using associated minisatellite diversity. These analyses suggest that the higher information content and sampling depth of these compound haplotypes may allow more precise calibration of lineage ages than is possible using coalescent analysis of DNA sequence. Using this method we have dated the oldest Eurasian lineage as 52,000-66,000 years and the oldest European specific lineage as 37,600-56,200 years. PMID- 11063727 TI - Expression of mutant alpha-synuclein causes increased susceptibility to dopamine toxicity. AB - Mutations of the alpha-synuclein gene have been identified in autosomal dominant Parkinson's disease (PD). Transgenic mice overexpressing wild-type human alpha synuclein develop motor impairments, intraneuronal inclusions and loss of dopaminergic terminals in the striatum. To study the mechanism of action through which mutant alpha-synuclein toxicity is mediated, we have generated stable, inducible cell models expressing wild-type or PD-associated mutant (G209A) alpha synuclein in human-derived HEK293 cells. Increased expression of either wild-type or mutant alpha-synuclein resulted in the formation of cytoplasmic aggregates which were associated with the vesicular (including monoaminergic) compartment. Expression of mutant alpha-synuclein induced a significant increase in sensitivity to dopamine toxicity compared with the wild-type protein expression. These results provide an explanation for the preferential dopaminergic neuronal degeneration seen in both the PD G209A mutant alpha-synuclein families and suggest that similar mechanisms may underlie or contribute to cell death in sporadic PD. PMID- 11063728 TI - Sequence and functional comparison in the Beckwith-Wiedemann region: implications for a novel imprinting centre and extended imprinting. AB - The clustered organization of most imprinted genes in mammals suggests coordinated genetic and epigenetic control mechanisms. Comparisons between human and mouse will help in elucidating these mechanisms by identifying structural and functional similarities. Previously we reported on such a comparison in the central part of the mouse imprinting cluster on distal chromosome 7 with the homologous Beckwith-Wiedemann syndrome (BWS) gene cluster on human chromosome 11p15.5. Here we focus on the adjacent sequences of 0.5 Mb including the KCNQ1/Kcnq1 and CDKN1C/Cdkn1c genes, which are implicated in BWS, and on one of the proposed boundary regions of the imprinting cluster. As in the previously analysed central region, this part of the cluster exhibits a highly conserved arrangement and structure of genes. The most striking similarity is found in the 3' part of the KCNQ1/Kcnq1 genes in large stretches of mostly non-coding sequences. The conserved region includes the recently identified KCNQ1OT1/Kcnq1ot1 antisense transcripts, flanked by a strikingly conserved cluster of LINE/Line elements and a CpG island which we show to carry a maternal germline methylation imprint. This region is likely to be the proposed second imprinting centre (IC2) in the BWS cluster. We also identified several novel genes inside and outside the previously proposed boundaries of the imprinting cluster. One of the genes outside the cluster, Obph1, is imprinted in mouse placenta indicating that at least in extra-embryonic tissues the imprinting cluster extends into a larger domain. PMID- 11063729 TI - Sequence interruptions confer differential stability at microsatellite alleles in mismatch repair-deficient cells. AB - Determinants of instability at a given microsatellite repeat merits investigation in view of relevance to understanding evolution of mutations at such sequences in human populations. The microsatellite D2S123 was studied as a paradigm CA repeat marker. Furthermore, this marker is one of a recommended panel used in molecular screening for hereditary non-polyposis colorectal cancer (HNPCC). In this investigation we show that the mutation rate at the D2S123 locus is markedly influenced by intra-allelic sequence variation within the repetitive tract itself. We employed a novel approach to characterize the nature of instability at D2S123, by utilizing cells derived from a non-tumour lineage, which harbour a dominant negative mismatch repair (MMR) mutation and a mutator phenotype. Individual alleles were typed using a semi-quantitative small pool PCR technique and this demonstrated substantial allele-these specific bias in susceptibility to mutation at the D2S123 locus. In support of these in vitro data, bias in allele mutation rate was also observed in tumours from 41 HNPCC patients, which was dependent on constitutional genotype. Sequencing of cell line and patient DNAs revealed that short alleles are significantly more susceptible to mutation due to the presence of uninterrupted CA repeats. Long D2S123 alleles are intrinsically more stable because of a TA interspersion within the repetitive tract. In addition to extending understanding of mutation at CA repeat dinucleotide tracts, these findings have considerable relevance both to screening programmes and to correlation of microsatellite instability (MSI) with colon cancer survival. The manifestation of tumour MSI may be substantially influenced by constitutional genotype. PMID- 11063730 TI - Novel mutations in lysosomal neuraminidase identify functional domains and determine clinical severity in sialidosis. AB - Lysosomal neuraminidase is the key enzyme for the intralysosomal catabolism of sialylated glycoconjugates and is deficient in two neurodegenerative lysosomal disorders, sialidosis and galactosialidosis. Here we report the identification of eight novel mutations in the neuraminidase gene of 11 sialidosis patients with various degrees of disease penetrance. Comparison of the primary structure of human neuraminidase with the primary and tertiary structures of bacterial sialidases indicated that most of the single amino acid substitutions occurred in functional motifs or conserved residues. On the basis of the subcellular distribution and residual catalytic activity of the mutant neuraminidases we assigned the mutant proteins to three groups: (i) catalytically inactive and not lysosomal; (ii) catalytically inactive, but localized in lysosome; and (iii) catalytically active and lysosomal. In general, there was a close correlation between the residual activity of the mutant enzymes and the clinical severity of disease. Patients with the severe infantile type II disease had mutations from group I, whereas patients with a mild form of type I disease had at least one mutation from group III. Mutations from the second group were mainly found in juvenile type II patients with intermediate clinical severity. Overall, our findings explain the clinical heterogeneity observed in sialidosis and may help in the assignment of existing or new allelic combinations to specific phenotypes. PMID- 11063731 TI - Alu-mediated PCR artifacts and the constitutional t(11;22) breakpoint. AB - The breakpoints of the recurrent t(11;22)(q23;q11) have recently been cloned. We identified palindromic AT-rich repeats (PATRRs) on 11q23 and 22q11 as the mechanism responsible for the rearrangement. Contradictory to our results, A.S. Hill et al. (Hum. Mol. Genet., 9, 1525-1532) suggested that Alu-mediated recombination is responsible. To clarify this discrepancy, the cloned 4.5 kb der(11) junction fragment has been completely sequenced. This sequence has been compared with that of an inverse PCR-generated der(11) junction fragment obtained by Hill et al. This reveals that the inverse PCR product has sustained a deletion between two Alu elements, such that the true breakpoint region is deleted from the PCR product. Utilizing PCR primers designed by Hill et al. to amplify across the der(11) breakpoint, we obtained a deleted PCR product even when our cloned der(11) junction fragment was used as template. Further, we find that the PCR primers that they utilized for amplification of the der(22) junction fragment are not located on the der(22). They are oriented in opposite directions within the region deleted from the der(11) PCR product, generating an artifact derived from the der(11) chromosome. Analysis of the truncated PCR products indicates a mixture of sequences from two distinct Alu elements, suggesting that the putative junction fragment described by Hill et al. is an Alu-mediated PCR artifact. These data suggest that caution should be exercised when analyzing PCR-based data, particularly when amplification is carried out in a region containing repeat structures with specific, difficult-to-amplify sequences. PMID- 11063732 TI - A novel frameshift mutation of the mtDNA COIII gene leads to impaired assembly of cytochrome c oxidase in a patient affected by Leigh-like syndrome. AB - We report on a novel frameshift mutation in the mtDNA gene encoding cytochrome c oxidase (COX) subunit III. The proband is an 11-year-old girl with a negative family history and an apparently healthy younger brother. Since 4 years of age, she has developed a progressive spastic paraparesis associated with ophthalmoparesis and moderate mental retardation. The presence of severe lactic acidosis and Leigh-like lesions of putamina prompted us to perform muscle and skin biopsies. In both, a profound, isolated defect of COX was found by histochemical and biochemical assays. Sequence analysis of muscle mtDNA resulted in the identification of a virtually homoplasmic frameshift mutation in the COIII gene, due to the insertion of an extra C at nucleotide position 9537 of mtDNA. Although the 9537C(ins) does not impair transcription of COIII, no full-length COX III protein was detected in mtDNA translation assays in vivo. Western blot analysis of two-dimensional blue-native electrophoresis showed a reduction of specific crossreacting material and the accumulation of early-assembly intermediates of COX, whereas the fully assembled complex was absent. One of these intermediates had an electrophoretic mobility different from those seen in controls, suggesting the presence of a qualitative abnormality of COX assembly. Immunostaining with specific antibodies failed to detect the presence of several smaller subunits in the complex lacking COX III, in spite of the demonstration that these subunits were present in the crude mitochondrial fraction of patient's cultured fibroblasts. Taken together, the data indicate a role for COX III in the incorporation and maintenance of smaller COX subunits within the complex. PMID- 11063733 TI - Polycystin-1, the product of the polycystic kidney disease 1 gene, co-localizes with desmosomes in MDCK cells. AB - Polycystin-1 is a novel protein predicted to be a large membrane-spanning glycoprotein with an extracellular N-terminus and an intracellular C-terminus, harboring several structural motifs. To study the subcellular localization, antibodies raised against various domains of polycystin-1 and against specific adhesion complex proteins were used for two-color immunofluorescence staining. In Madine Darby canine kidney (MDCK) cells, polycystin-1 was detected in the cytoplasm as well as co-localizing with desmosomes, but not with tight or adherens junctions. Using confocal laser scanning and immunoelectron microscopy we confirmed the desmosomal localization. By performing a calcium switch experiment, we demonstrated the sequential reassembly of tight junctions, subsequently adherens junctions and finally desmosomes. Polycystin-1 only stained the membrane after incorporation of desmoplakin into the desmosomes, suggesting that membrane-bound polycystin-1 may be important for cellular signaling or cell adhesion, but not for the assembly of adhesion complexes. PMID- 11063734 TI - Surfactant proteins A and B as interactive genetic determinants of neonatal respiratory distress syndrome. AB - Prematurity is the most important risk factor predisposing to neonatal respiratory distress syndrome (RDS). Genetic factors are likely to contribute to the risk of this complex disease. The present study was designed to investigate whether the surfactant protein B (SP-B) gene or interaction between the SP-A and SP-B genes has a role in the genetic susceptibility to RDS. The genotype analyses were performed on 684 prematurely born neonates, of whom 184 developed RDS. Of the two SP-B polymorphisms genotyped, the Ile131Thr variation affects a putative N-terminal N:-linked glycosylation site of proSP-B and the length variation of intron 4 has previously been suggested to associate with RDS. Neither of the two SP-B polymorphisms associated directly with RDS or with prematurity. Instead, our data show that the previously identified association between SP-A alleles and RDS was dependent on the SP-B Ile131Thr genotype. On the basis of chi(2) and logistic regression analyses, the SP-A allele, haplotype and genotype distributions differed significantly between the RDS infants and controls only when the SP-B genotype was Thr/Thr. Among the infants born before 32 weeks of gestation and having the SP-B genotype Thr/Thr, the SP-A1 allele 6A(2) was over-represented in RDS group compared with controls (P = 0.001, OR = 4.7, CI 1.8-12.2). In the same comparison, the SP-A1 allele 6A(3) was under-represented in RDS (P = 0.001, OR = 0.2, CI 0.1-0.6). We propose that the SP-B Ile131Thr polymorphism is a determinant for certain SP-A alleles as factors causing genetic susceptibility to RDS (6A(2), 1A(0)) or protection against it (6A(3), 1A(2)). PMID- 11063735 TI - Recessive mutation in desmoplakin disrupts desmoplakin-intermediate filament interactions and causes dilated cardiomyopathy, woolly hair and keratoderma. AB - Desmosomes are major cell adhesion junctions, particularly prominent in the epidermis and cardiac tissue and are important for the rigidity and strength of the cells. The desmosome consists of several proteins, of which desmoplakin is the most abundant. Here, we describe the first recessive human mutation, 7901delG, in the desmoplakin gene which causes a generalized striate keratoderma particularly affecting the palmoplantar epidermis, woolly hair and a dilated left ventricular cardiomyopathy. A number of the patients with this syndromic disorder suffer heart failure in their teenage years, resulting in early morbidity. All tested affected members of three families from Ecuador were homozygous for this mutation which produces a premature stop codon leading to a truncated desmoplakin protein missing the C domain of the tail region. Histology of the skin revealed large intercellular spaces and clustering of desmosomes at the infrequent sites of keratinocyte adhesion. Immunohistochemistry of skin from the patients showed a perinuclear localization of keratin in suprabasal keratinocytes, suggesting a collapsed intermediate filament network. This study demonstrates the importance of desmoplakin in the attachment of intermediate filaments to the desmosome. In contrast to null DESMOPLAKIN: mice which die in early development, the truncated protein due to the homozygous 7901delG mutation in humans is not embryonic lethal. This suggests that the tail domain of desmoplakin is not required for establishing tissue architecture during development. PMID- 11063736 TI - Gender of the embryo contributes to CAG instability in transgenic mice containing a Huntington's disease gene. AB - Gender is known to influence the transmission of trinucleotide repeats in human disease. However, the molecular basis for the parent-of-origin effect associated with trinucleotide repeat expansion is not known. We have followed, during transmission, the fate of the CAG trinucleotide repeat in a transgene containing the exon 1 portion of the human Huntington's disease (HD) gene. Similar to humans, the mouse transmits expansions predominantly through the male germ line. Surprisingly, we find that the CAG repeat size of the mutant human HD gene is different in male and female progeny from identical fathers. Males predominantly expand the repeat whereas females predominantly contract the repeat. In contrast to the classic definition of imprinting, CAG expansion is influenced by the gender of the embryo. Our results raise the possibility that there are X- or Y encoded factors that influence repair or replication of DNA in the embryo. Gender dependence in the embryo may explain why expansion in HD from premutation to disease primarily occurs through the paternal line. PMID- 11063737 TI - Mutations in yeast ARV1 alter intracellular sterol distribution and are complemented by human ARV1. AB - Intracellular cholesterol redistribution between membranes and its subsequent esterification are critical aspects of lipid homeostasis that prevent free sterol toxicity. To identify genes that mediate sterol trafficking, we screened for yeast mutants that were inviable in the absence of sterol esterification. Mutations in the novel gene, ARV1, render cells dependent on sterol esterification for growth, nystatin-sensitive, temperature-sensitive, and anaerobically inviable. Cells lacking Arv1p display altered intracellular sterol distribution and are defective in sterol uptake, consistent with a role for Arv1p in trafficking sterol into the plasma membrane. Human ARV1, a predicted sequence ortholog of yeast ARV1, complements the defects associated with deletion of the yeast gene. The genes are predicted to encode transmembrane proteins with potential zinc-binding motifs. We propose that ARV1 is a novel mediator of eukaryotic sterol homeostasis. PMID- 11063739 TI - Direct interaction of Rab4 with syntaxin 4. AB - In the present study, we examined the possible interaction between Rab4 and syntaxin 4, both having been implicated in insulin-induced GLUT4 translocation. Rab4 and syntaxin 4 were coimmunoprecipitated from the lysates of electrically permeabilized rat adipocytes. The interaction between the two proteins was reduced by insulin treatment and increased by the addition of guanosine 5'-O-(3 thiotriphosphate) (GTPgammaS). An in vitro binding assay revealed that the bacterially expressed Rab4 was bound to a glutathione S-transferase fusion protein containing the cytoplasmic domain of syntaxin 4 (GST-syntaxin 4-(1-273)) but not to syntaxin 1A or vesicle-associated membrane protein-2. The interaction between Rab4 and syntaxin 4 seemed to be regulated by the guanine nucleotide status of Rab4, because 1) GTPgammaS treatment of the cells significantly increased, but guanosine 5'-O-(2-thiodiphosphate) (GDPbetaS) treatment decreased the amount of Rab4 pulled down with GST-syntaxin 4-(1-273) from the cell lysates; 2) GTPgammaS loading on Rab4 caused a marked increase in the affinity of Rab4 to syntaxin 4 whereas GDPbetaS loading had little effect; and 3) a GTPase-deficient mutant of Rab4 (Rab4(Q67L)), but not a GTP-binding-defective mutant (Rab4(S22N)), was bound to GST-syntaxin 4-(1-273). Although insulin stimulated [gamma-(32)P]GTP binding to Rab4 in a time-dependent fashion, its effect on the Rab4 interaction with syntaxin 4 was apparently biphasic; an initial increase in Rab4 associated with syntaxin 4 was followed by a gradual dissociation of the GTPase from syntaxin 4. Finally, the binding of Rab4(Q67L) to GST-syntaxin 4-(1-273) was inhibited by munc-18c in a dose-dependent manner, indicating that GTP-loaded Rab4 binds to syntaxin 4 in the open conformation. These results suggest that 1) Rab4 interacts with syntaxin 4 in a direct and specific manner, and 2) the interaction is regulated by the guanine nucleotide status of Rab4 as well as by the conformational status of syntaxin 4. PMID- 11063740 TI - Glycogen synthase kinase-3 inhibits the DNA binding activity of NFATc. AB - The NFAT family of transcription factors is required for the expression of numerous immunologically important genes and plays a pivotal role in both the initiation and coordination of the immune response. NFAT family members appear to be regulated primarily at the level of their subcellular localization. Here we show that NFATc is additionally regulated at the level of its DNA binding activity. Using gel mobility shift assays, we demonstrate that the intrinsic DNA binding activity of NFATc is negatively regulated by phosphorylation. We found that activation of calcineurin activity in cells and dephosphorylation of NFATc in vitro enhanced NFATc DNA binding activity, whereas phosphorylation of NFATc in vitro inhibited its ability to bind DNA. Through the analysis of NFATc mutants, we identified the conserved Ser-Pro repeat motifs as critical quantitative determinants of NFATc DNA binding activity. In addition, we provide several lines of evidence to suggest that the phosphorylation of the Ser-Pro repeats by glycogen synthase kinase-3 inhibits the ability of NFATc to bind DNA. Taken together, these studies afford new insights into the regulation of NFATc and underscore the potential role of glycogen synthase kinase-3 in the regulation of NFAT-dependent gene expression. PMID- 11063741 TI - Translation initiation of the insulin-like growth factor I receptor mRNA is mediated by an internal ribosome entry site. AB - The insulin-like growth factor I receptor (IGF-IR) is a heterotetrameric receptor mediating the effects of insulin-like growth I and other growth factors. This receptor is encoded by an mRNA containing an unusually long, G-C-rich, and highly structured 5' untranslated region. Using bicistronic constructs, we demonstrated here that the 5' untranslated region of the IGF-IR allows translation initiation by internal ribosome entry and therefore constitutes an internal ribosome entry site. In vitro cross-linking revealed that this internal ribosome entry site binds a protein of 57 kDa. Immunoprecipitation of UV cross-linked proteins proved that this protein was the polypyrimidine tract-binding protein, a well known regulator of picornavirus mRNA translation. The efficiency of translation of the endogenous IGF-IR mRNA is not affected by rapamycin, which is a potent inhibitor of cap-dependent translation. This result provides evidence that the endogenous IGF-IR mRNA is translated, at least in part, through a cap-independent mechanism. This is the first report of a growth factor receptor containing sequence elements that allow translation initiation to occur by internal initiation. Because the IGF-IR has a pivotal function in the cell cycle, this mechanism of translation regulation could play a crucial role in the control of cell proliferation and differentiation. PMID- 11063742 TI - c-Jun NH2-terminal kinase-mediated redox-dependent degradation of IkappaB: role of thioredoxin in NF-kappaB activation. AB - NF-kappaB is a redox-sensitive transcription factor known to be activated by oxidative stress as well as chemical and biological reductants. Its DNA binding activity requires reduced cysteines present in the p65 subunit of the dimer. Thioredoxin (Trx) is an endogenous disulfide oxidoreductase known to modulate several redox-dependent functions in the cell. NF-kappaB was activated by addition of Escherichia coli thioredoxin in a redox-dependent manner in A549 cells. Such activation was accompanied by degradation of IkappaB in the cytosol. In addition, only the reduced form of thioredoxin activated NF-kappaB, whereas the oxidized form was without any effect. Overexpression of human thioredoxin also caused activation of NF-kappaB and degradation of IkappaB. On the contrary, dominant-negative redox-inactive mutant thioredoxin expression did not activate NF-kappaB, further confirming the redox-dependent activation of NF-kappaB. We also investigated the mechanism of activation of NF-kappaB by thioredoxin. We demonstrate that thioredoxin activates c-Jun NH(2)-terminal kinase (JNK) signaling cascade, and dominant-negative expression of mitogen-activated protein kinase kinase kinase 1 (MEKK1), JNK kinase, or JNK inhibits NF-kappaB activation by thioredoxin. In contrast, wild-type MEKK1 or JNK kinase induced NF-kappaB activation alone or in combination with thioredoxin expression plasmid. These findings were also confirmed by NF-kappaB-dependent luciferase reporter gene transcription. PMID- 11063743 TI - Apoptosis-resistant mitochondria in T cells selected for resistance to Fas signaling. AB - Jurkat leukemic T cells are highly sensitive to the extrinsic pathways of apoptosis induced via the death receptor Fas or tumor necrosis factor-related apoptosis-inducing ligand as well as to the intrinsic/mitochondrial pathways of death induced by VP-16 or staurosporin. We report here that clonal Jurkat cell lines selected for resistance to Fas-induced apoptosis were cross-resistant to VP 16 or staurosporin. Each of the apoptotic pathways was blocked at an apical phase, where common regulators of apoptosis have not yet been defined. The Fas pathway was blocked at the level of caspase-8, whereas the intrinsic pathway was blocked at the mitochondria. No processing or activity of caspases was detected in resistant cells in response to either Fas-cross-linking or VP-16 treatment. Also, no apoptosis-associated alterations in the mitochondrial inner membrane, outer membrane, or matrix were detected in resistant Jurkat cells treated with VP 16. Thus, no changes in permeability transition, loss in inner membrane cardiolipin, generation of reactive oxygen species, or release of cytochrome c were observed in resistant cells treated with VP-16. Further, unlike purified mitochondria from wild type cells, those obtained from resistant cells did not release cytochrome c or apoptosis-inducing factor in response to recombinant Bax or truncated Bid. These results identify a defect in mitochondria ability to release intermembrane proteins in response to Bid or Bax as a mechanism of resistance to chemotherapeuetic drugs. Further, the selection of VP-16-resistant mitochondria via elimination of Fas-susceptible cells may suggest the existence of a shared regulatory component between the extrinsic and intrinsic pathways of apoptosis. PMID- 11063744 TI - Protein kinase C-zeta phosphorylates insulin receptor substrate-1 and impairs its ability to activate phosphatidylinositol 3-kinase in response to insulin. AB - Protein kinase C-zeta (PKC-zeta) is a serine/threonine kinase downstream from phosphatidylinositol 3-kinase in insulin signaling pathways. However, specific substrates for PKC-zeta that participate in the biological actions of insulin have not been reported. In the present study, we identified insulin receptor substrate-1 (IRS-1) as a novel substrate for PKC-zeta. Under in vitro conditions, wild-type PKC-zeta (but not kinase-deficient mutant PKC-zeta) significantly phosphorylated IRS-1. This phosphorylation was reversed by treatment with the serine-specific phosphatase, protein phosphatase 2A. In addition, the overexpression of PKC-zeta in NIH-3T3(IR) cells caused significant phosphorylation of cotransfected IRS-1 as demonstrated by [(32)P]orthophosphate labeling experiments. In rat adipose cells, endogenous IRS-1 coimmunoprecipitated with endogenous PKC-zeta, and this association was increased 2-fold upon insulin stimulation. Furthermore, the overexpression of PKC-zeta in NIH-3T3(IR) cells significantly impaired insulin-stimulated tyrosine phosphorylation of cotransfected IRS-1. Importantly, this was accompanied by impaired IRS-1 associated phosphatidylinositol 3-kinase activity. Taken together, our results raise the possibility that IRS-1 is a novel physiological substrate for PKC-zeta. Because PKC-zeta is located downstream from IRS-1 and phosphatidylinositol 3 kinase in established insulin signaling pathways, PKC-zeta may participate in negative feedback pathways to IRS-1 similar to those described previously for Akt and GSK-3. PMID- 11063745 TI - Synthesis and characterization of insulin-like growth factor (IGF)-1 photoprobes selective for the IGF-binding proteins (IGFBPS). photoaffinity labeling of the IGF-binding domain on IGFBP-2. AB - Elevated insulin-like growth factor (IGF)-1 levels are prognostic for the development of prostate and breast cancers and exacerbate the complications of diabetes. In each case, perturbation of the balance between IGF-1/2, the IGF-1 receptor, and the IGF-binding proteins (IGFBPs) leads to elevated IGF-1 sensitivity. Blockade of IGF action in these diseases would be clinically significant. Unfortunately, effective IGF antagonists are currently unavailable. The IGFBPs exhibit high affinity and specificity for the IGFs and serve as natural IGF antagonists, limiting their mitogenic/anti-apoptotic effects. As an initial step in designing IGFBP-based agents that antagonize IGF action, we have begun to analyze the structure of the IGF-binding site on IGFBP-2. To this end, two IGF-1 photoprobes, N(alphaGly1)-(4-azidobenzoyl)-IGF-1 (abG(1)IGF-1) and N(alphaGly1)-([2-6-(biotinamido)-2(p-azidobenzamido)hexanoamido]ethyl-1,3' dithiopropionoyl)-IGF-1 (bedG(1)IGF-1), selective for the IGFBPs were synthesized by derivatization of the alpha-amino group of Gly(1), known to be part of the IGFBP-binding domain. Mass spectrometric analysis of the reduced, alkylated, and trypsin-digested abG(1)IGF-1.recombinant human IGFBP-2 (rhIGFBP-2) complex indicated photoincorporation near the carboxyl terminus of rhIGFBP-2, between residues 266 and 287. Mass spectrometric analysis of avidin-purified tryptic peptides of the bedG(1)IGF-1.rhIGFBP-2 complex revealed photoincorporation within residues 212-227. Taken together, these data indicate that the IGFBP-binding domain on IGF-1 contacts the distal third of IGFBP-2, providing evidence that the IGF-1-binding domain is located within the C terminus of IGFBP-2. PMID- 11063746 TI - Protein kinase C phosphorylates RGS2 and modulates its capacity for negative regulation of Galpha 11 signaling. AB - RGS proteins (regulators of G protein signaling) attenuate heterotrimeric G protein signaling by functioning as both GTPase-activating proteins (GAPs) and inhibitors of G protein/effector interaction. RGS2 has been shown to regulate Galpha(q)-mediated inositol lipid signaling. Although purified RGS2 blocks PLC beta activation by the nonhydrolyzable GTP analog guanosine 5'-O-thiophosphate (GTPgammaS), its capacity to regulate inositol lipid signaling under conditions where GTPase-promoted hydrolysis of GTP is operative has not been fully explored. Utilizing the turkey erythrocyte membrane model of inositol lipid signaling, we investigated regulation by RGS2 of both GTP and GTPgammaS-stimulated Galpha(11) signaling. Different inhibitory potencies of RGS2 were observed under conditions assessing its activity as a GAP versus as an effector antagonist; i.e. RGS2 was a 10-20-fold more potent inhibitor of aluminum fluoride and GTP-stimulated PLC betat activity than of GTPgammaS-promoted PLC-betat activity. We also examined whether RGS2 was regulated by downstream components of the inositol lipid signaling pathway. RGS2 was phosphorylated by PKC in vitro to a stoichiometry of approximately unity by both a mixture of PKC isozymes and individual calcium and phospholipid-dependent PKC isoforms. Moreover, RGS2 was phosphorylated in intact COS7 cells in response to PKC activation by 4beta-phorbol 12beta-myristate 13alpha-acetate and, to a lesser extent, by the P2Y(2) receptor agonist UTP. In vitro phosphorylation of RGS2 by PKC decreased its capacity to attenuate both GTP and GTPgammaS-stimulated PLC-betat activation, with the extent of attenuation correlating with the level of RGS2 phosphorylation. A phosphorylation-dependent inhibition of RGS2 GAP activity was also observed in proteoliposomes reconstituted with purified P2Y(1) receptor and Galpha(q)betagamma. These results identify for the first time a phosphorylation-induced change in the activity of an RGS protein and suggest a mechanism for potentiation of inositol lipid signaling by PKC. PMID- 11063747 TI - Hormone status selects for spontaneous somatic androgen receptor variants that demonstrate specific ligand and cofactor dependent activities in autochthonous prostate cancer. AB - We have used the autochthonous transgenic adenocarcinoma of mouse prostate (TRAMP) model to investigate the relationship between somatic mutation in the androgen receptor (AR) and the emergence of androgen-independent prostate cancer. Here we report the identification, isolation, and characterization of distinct classes of AR variants from spontaneous prostate tumors in the TRAMP model. Using cDNA cloning, single stranded conformation polymorphism and sequencing strategies, 15 unique somatic mutations in the AR were identified in prostate tumors obtained from eight TRAMP mice between 24 and 29 weeks of age. At least one mutation was isolated from each mouse. All mutations were single base substitutions, 10 were missense and 5 were silent. Nine mutations in the AR were identified in tumors of four mice that were castrated at 12 weeks of age. Interestingly, the majority of mutations (seven out of nine, 78%) identified in the androgen-independent tumors colocalized in the AR transactivation domain. The remaining mutations colocalized in the AR ligand binding domain. In general, the AR variants demonstrated promoter-, cell-, and cofactor-specific activities in response to various hormones. All AR variants isolated in this study maintained strong sensitivity for androgens, and four AR variants isolated from castrated mice demonstrated increased activities in the absence of ligand. The K638M and F677S variants demonstrated increased activities in response to androgen, and K638M also demonstrated increased response to estradiol. In the presence of AR coactivator ARA70 the E231G variant demonstrated increased activity in response to both androgen and estradiol. However, in the presence of AR coactivator ARA160 the E231G variant was selectively responsive to androgen. Collectively these analyses not only indicate that somatic mutations in the AR gene occur spontaneously in TRAMP tumors but also how changes in the hormonal environment may drive the selection of spontaneous somatic mutations that provide a growth advantage. PMID- 11063748 TI - Insights into ligand binding and catalysis of a central step in NAD+ synthesis: structures of Methanobacterium thermoautotrophicum NMN adenylyltransferase complexes. AB - Nicotinamide mononucleotide adenylyltransferase (NMNATase) catalyzes the linking of NMN(+) or NaMN(+) with ATP, which in all organisms is one of the common step in the synthesis of the ubiquitous coenzyme NAD(+), via both de novo and salvage biosynthetic pathways. The structure of Methanobacterium thermoautotrophicum NMNATase determined using multiwavelength anomalous dispersion phasing revealed a nucleotide-binding fold common to nucleotidyltransferase proteins. An NAD(+) molecule and a sulfate ion were bound in the active site allowing the identification of residues involved in product binding. In addition, the role of the conserved (16)HXGH(19) active site motif in catalysis was probed by mutagenic, enzymatic and crystallographic techniques, including the characterization of an NMN(+)/SO4(2-) complex of mutant H19A NMNATase. PMID- 11063749 TI - Molecular mechanism of hypoxia-inducible factor 1alpha -p300 interaction. A leucine-rich interface regulated by a single cysteine. AB - Hypoxia-inducible factor 1alpha (HIF1alpha) plays a pivotal role in embryogenesis, angiogenesis, and tumorigenesis. HIF1alpha-mediated transcription requires the coactivator p300, at least in part, through interaction with the cysteine- and histidine-rich 1 domain of p300. To understand the molecular basis of this interaction, we have developed a random mutagenesis screen in yeast approach for efficient identification of residues that are functionally critical for protein interactions. As a result, four residues (Leu-795, Cys-800, Leu-818, and Leu-822) in the C-terminal activation domain of HIF1alpha have been identified as crucial for HIF1 transactivation in mammalian systems. Moreover, data from residue substitution experiments indicate the stringent necessity of leucine and hydrophobic cysteine for C-terminal activation domain function. Likewise, Leu-344, Leu-345, Cys-388, and Cys-393 in the cysteine- and histidine rich 1 domain of p300 have also been shown to be essential for the functional interaction. We propose that hypoxia-induced HIF1alpha-p300 interaction relies upon a leucine-rich hydrophobic interface that is regulated by the hydrophilic and hydrophobic sulfhydryls of HIF1alpha Cys-800. PMID- 11063751 TI - Requirement of Ras/MAPK pathway activation by transforming growth factor beta for transforming growth factor beta 1 production in a smad-dependent pathway PMID- 11063750 TI - Role of the C-terminal G3 domain in sorting and secretion of aggrecan core protein and ubiquitin-mediated degradation of accumulated mutant precursors. AB - Aggrecan is a complex multidomain macromolecule that undergoes extensive processing and post-translational modification. A thorough understanding of the events and signals that promote translocation of aggrecan through the secretory pathway is lacking. To investigate which features of the C-terminal G3 region are necessary for successful translocation of the core protein, a number of deletion constructs based on the chick aggrecan cDNA sequence were prepared and transiently expressed in COS-1 cells and the natural host, embryonic chick chondrocytes; stable cell lines were established as well. The present results clearly establish a precise requirement for that portion of the G3 C-lectin domain encoded by exon 15 for: (i) translocation from the endoplasmic reticulum (ER) to the Golgi, (ii) secretion from the cell, (iii) galactosylation of chondroitin sulfate (CS) chains, (iv) generation of Ca(+2)-dependent galactose binding ability. Furthermore, in the absence of this subdomain there is excess accumulation in the ER of expression products leading to a stress-related response involving the chaperones Grp78 and protein disulfide isomerase, followed by degradation via a ubiquitin-proteosome pathway. All of these events in the model system faithfully mimic the naturally occurring nanomelic mutant, which also elicits a ubiquitin-mediated degradation response due to the accumulation of the truncated core protein precursor. This study represents the first report of the mode of degradation of overexpressed or misfolded proteoglycans and suggests that, although proteoglycans follow different glycosylation pathways from other glycoproteins, they are monitored by an ER surveillance system similar to that which detects other misfolded proteins. PMID- 11063752 TI - The role of gamma A/gamma ' fibrinogen in plasma factor XIII activation PMID- 11063753 TI - The role of positively charged amino acids in ATP recognition by human P2X1 receptors PMID- 11063754 TI - Loss of cone molecular markers in rhodopsin-mutant human retinas with retinitis pigmentosa. AB - PURPOSE: To examine the effect of rhodopsin mutations on cone photoreceptors in human retinas with retinitis pigmentosa (RP). METHODS: Four RP retinas with rhodopsin mutations and four normal retinas were examined by immunofluorescence with a battery of cell-specific antibodies against cone and rod cytoplasmic and outer segment membrane proteins. Areas of the retinas were studied that showed maximal preservation of photoreceptor structure. RESULTS: All four RP retinas showed loss of rods, ranging from mild (T-17-M), to more severe (P-23-H), to advanced degeneration (Q-64-ter and G-106-R). The majority of cones in the T-17-M and P-23-H retinas were cytologically normal but showed loss of immunoreactivity for the cytoplasmic proteins 7G6, calbindin, and X-arrestin. The cone outer segments (OS) remained positive for cone opsins and peripherin-2 (rds/peripherin). All remaining cones in the Q-64-ter and G-106-R retinas were degenerate, with short to absent OS, but had strong reactivity for these cytoplasmic and OS membrane markers. Cones in the maculas of the RP retinas were degenerate, with short to absent OS, but retained strong labeling for the cytoplasmic and OS proteins. CONCLUSIONS: Even before cones show cytologic changes in response to rod cell degeneration, they lose immunoreactivity for certain cytoplasmic proteins. These cones later show shortening and loss of OS, although their OS membrane proteins remain well labeled. Cones may down regulate expression of both cytoplasmic and outer segment membrane proteins in response to mutant rod cell dysfunction and/or cell death in human RP retinas. Such cytologic and immunocytochemical changes in the cones may presage death of these critical cells in the later stages of RP. PMID- 11063755 TI - Tyrosine phosphorylation is involved in phosphatidylinositol 3-kinase activation in bovine rod outer segments. AB - PURPOSE: We have previously shown that phosphatidylinositol 3-kinase (PI 3 kinase) activity is present in bovine rod outer segments (ROS). The present study was undertaken to investigate the mechanism of PI 3-kinase activation in these membranes. METHODS: Tyrosine-phosphorylated ROS (PY-ROS) were obtained by incubating ROS with ATP, MgCl2, and orthovanadate (Na3VO4), a tyrosine phosphatase inhibitor. Non-phosphorylated ROS (N-ROS) were obtained by incubating ROS under the same conditions, but without ATP and orthovanadate. Both were subjected to immunoprecipitation using antibodies against the regulatory p85 (anti-p85) subunit of PI 3-kinase, the catalytic p110 (anti-p110) subunit of PI 3 kinase, or phosphotyrosine (anti-PY). The immunoprecipitates (IPs) were assayed for PI 3-kinase activity. Enzyme assay products were separated by thin-layer chromatography (TLC), deacylated, and identified by high performance liquid chromatography (HPLC). RESULTS: PI 3-kinase activity in anti-p85 and p110alpha IPs was significantly higher in PY-ROS than in N-ROS. No enzyme activity was recovered in anti-p110beta IPs. PI 3-kinase activity in anti-PY IPs from PY-ROS was six-fold greater than those from N-ROS. Immunoblot analysis showed that the amount of p85 in PY IPs from PY-ROS was significantly higher than those from N ROS. However, tyrosine phosphorylation of p85 and p110alpha was not observed in anti-p85 and anti-p110alpha IPs that were probed with anti-PY. CONCLUSIONS: This study indicates that the p85/p110alpha complex of PI 3-kinase is present in ROS and tyrosine phosphorylation is involved in the regulation of its activity. PMID- 11063756 TI - Medical school in Split: intentions and achievements. AB - This paper describes the origins, aims, and current status of the Split University School of Medicine. Split University School of Medicine was founded several times anew (in 1806, 1944, and 1974), and eventually started to operate as an independent faculty on March 26, 1997. Conceived as a small and efficient medical school, each year it enrolls 50 freshmen at the most. The whole curriculum contains 27 courses (no "majors" or "minors"), each of around 200 class hours, which amounts to a total of 5,610 teaching hours. The teaching process aims at achieving skills and knowledge necessary in general medical practice, at establishing an intellectual basis for further education, and corresponding with the specific medical needs of the coastal area ("Adriatic orientation") Student/instructor ratio is close to 2.0 and the teaching goals are defined better than in older medical schools in the region. The first results are already tangible. PMID- 11063757 TI - Use of Journal Citation Reports and Journal Performance Indicators in measuring short and long term journal impact. AB - The impact factor has become the subject of widespread controversy. It has gradually developed to mean both journal and author impact. The emphasis on impact factors obscures the main purpose of bibliographic databases created at the Institute for Scientific Information. I will here show how two of these databases, Journal Citation Reports and the Journal Performance Indicators, can be used to study scientific journals and the articles they publish, as well as the evolution of scientific fields. PMID- 11063758 TI - Priority setting, justice, and health care: conceptual analysis. AB - In this paper, priority setting in health care is defined as distributional decisions at the individual level involving clear and direct limitation of access to beneficial health care according to some categorical criterion other than the market. The justification of a particular rule of priority setting depends on conceptual issues as well as on the choice of value premises. Especially important is the type of scarcity involved (whether the imbalance between supply and need can be overcome or not) and the concept of justice used or presupposed (for instance, whether the conception of justice emphasizes efficiency or fairness or both). The proposal put forward is that the task of medical ethics is to provide conceptual clarification and value premises relevant to justify rules of priority setting. The actual choice of such rules belongs primarily to the domain of politics. PMID- 11063759 TI - Prevention of herpes genitalis by the 'Bulgarian' vaccine F.HSV-2V(PRK): preliminary clinical evidence. AB - AIM: To examine the antigenic properties of the formalin-inactivated herpes simplex virus type 2 (HSV-2) virus-particle vaccine F. HSV-2V(PRK), which has been used therapeutically in Bulgaria for 30 years, and to make preliminary assessment of its potential protective efficacy by a follow-up of vaccinated patients with herpes genitalis. METHODS: Properties of the vaccine were examined by standard immunological laboratory tests. Fifty-five patients at risk of herpes genitalis received 2-4 vaccinations and were monitored during a 6-year follow-up. RESULTS: The vaccine was antigenic in laboratory tests and absorbed neutralizing antibody from hyperimmune rabbit serum against herpes simplex virus type 1 (HSV 1). In vaccinated patients, there was an overall contraction rate of herpes genitalis of 5.4%. There was no evidence of significant local or generalized adverse effects from vaccination. CONCLUSION: Bulgarian vaccine F.HSV-2V(PRK) may have protective efficacy, which, in association with its apparent safety from our findings and from its clinical use for over 30 years in Bulgaria, suggests that it should be scrutinized by a formal clinical trial. PMID- 11063760 TI - Intramedullar stimulation of the facial and hypoglossal nerves: estimation of the stimulated site. AB - AIM: To determine the stimulation site of both facial and hypoglossal nerves after transcranial magnetic stimulation. METHODS: After surgical exposure of the brainstem in 22 patients with intrinsic pontine (n=9) or medullary (n=13) tumors, the facial colliculus and the hypoglossal triangle were electrically stimulated. The EMG responses were recorded with flexible wire electrodes from the orbicularis oculi/orbicularis oris muscles, and genioglossal muscles. Patients had no preoperative deficit of the nerves. RESULTS: The EMG mean latencies of the unaffected facial nerve were 5.2+/-0.6 ms for the orbicularis oculi, and 5.2+/ 0.5 ms for the orbicularis oris muscle. After the stimulation of 18 possibly affected facial nerves, the EMG mean latencies were 5.3+/-0.3 ms for the orbicularis oculi (p=0.539, unpaired Student's t-test), and 5.4+/-0.2 ms for the orbicularis oris (p=0.122). The EMG mean latency of the unaffected hypoglossal nerve was 4.1+/-0.6 ms for the genioglossal muscle. After the stimulation of 26 possibly affected hypoglossal nerves, the EMG mean latency for the genioglossal muscle was 5.3+/-0.3 ms. There was a significant difference (p<0.001) in latency for genioglossal EMG responses between the patients with pontine and those with medullary tumors. CONCLUSION: Shorter EMG mean latencies of unaffected facial nerves obtained after direct stimulation of the facial colliculi confirm that magnetic stimulation is most likely to occur closer to the nerve's exit from the brainstem than to its entrance into the internal auditory meatus. The hypoglossal nerve seems to have the site of excitation at the axon hillock of the hypoglossal motor neurons. PMID- 11063761 TI - Clinical variability and molecular diagnosis in a four-generation family with X linked Emery-Dreifuss muscular dystrophy. AB - AIM: To describe the clinical variability of X-linked Emery-Dreifuss muscular dystrophy (X-EDMD) with cardiac involvement in a four-generation family with a novel mutation in the STA gene. METHODS: Clinical data were provided for 4 affected males and a female carrier. The Western blot analysis of emerin was performed on lymphoblastoid cell lines and followed by sequencing of the emerin gene. RESULTS: A thymine insertion at nucleotide 417 in exon 2, resulting in a frameshift with a premature stop codon at position 62 and absence of functional protein, was found in one of the three available patients. In ten-year-old proband's dizygotic twin-nephews the intermittent first-degree A-V block, atrial and ventricular ectopy, atrial runs, and exit sinus block were found, although the echocardiographic findings were normal. One of the twins also had short episodes of atrial fibrillation, idioventricular rhythm, and junctional rhythm. CONCLUSION: Cardiac abnormalities in the proband's ten-year-old dizygotic twins without evident clinical features suggestive of EDMD were remarkable in contrast to the oldest patient in the family, who lived to the age of 63 without a pacemaker, and to the proband who had a very early onset of muscle wasting and weakness, and a pacemaker implantation at the age of 27. This striking intra familial variability in cardiac involvement associated with specific null mutation (417 ins T) has practical early diagnostic and possibly preventive implications. It also points at genetic and environmental factors as causes of clinical features in X-EDMD. PMID- 11063762 TI - Determinants of reduced bone mineral density and increased bone turnover after kidney transplantation: cross-sectional study. AB - AIM: To analyze bone metabolism and the risk factors of bone loss in kidney transplant recipients. METHODS: The bone mineral density (BMD) of the lumbar spine, femoral neck, and radius was determined by dual-energy X-ray absorptiometry in 52 patients 8 days to 228 months after kidney transplantation. Total and bone alkaline phosphatase (BAP), osteocalcin, procollagen, type I collagen telopeptide, collagen cross links, calcium, intact parathyroid hormone (iPTH), and creatinine were measured in all patients. RESULTS: The BMD of the spine and femoral neck was reduced in 57%, and of the radius in 72% of the patients. Reduced BMD was associated with significantly increased levels of iPTH, osteocalcin, and procollagen. Dialysis duration negatively correlated with the radius BMD in all patients and the femoral neck BMD in women. No relationship between BMD and length of post-transplantation time, age, cumulative steroid dose, or serum creatinine level was established. All biochemical parameters negatively correlated with the spine BMD, but not with the BMD of the femoral neck and radius. The correlation between BAP and telopeptide and length of post transplantation time was also negative. No difference in the incidence of osteopenia was found between genders. CONCLUSION: Osteopenia/osteoporosis and increased bone turnover were present in more than a half of the kidney transplant recipients. Reduced BMD was associated with enhanced bone remodeling, primarily mediated by PTH hypersecretion. The length of post-transplantation period, cumulative steroid dose, gender, and age could not be identified as risk factors of reduced BMD. PMID- 11063763 TI - Eligibility of patients with acute myocardial infarction for thrombolytic therapy: retrospective cohort study. AB - AIM: To investigate the eligibility of patients with acute myocardial infarction (AMI) for thrombolytic therapy (TT) and evaluate the results of treatment. METHODS: Retrospective analysis included 366 patients with AMI, mean age 66+/-11 years, treated in 1999. We analyzed age, gender, previous infarction, previous TT, present TT with streptokinase and its effects on the course and outcome, pain to-door time, and door-to-needle time. Reperfusion and reocclusion were evaluated non-invasively according to the occurrence of the reperfusion and reocclusion syndrome. RESULTS: One hundred patients (27%) underwent TT. It was less frequently applied in older patients, women, and patients with previous myocardial infarction. Reperfusion was achieved in 66 (66%) patients and reocclusion occurred in 9 (14%). Final outcome was successful in 57 (57%) patients. The TT group had more frequent arrhythmias (67% vs. 41%, p<0.001) and less frequent heart failure (20% vs. 39%, p<0.001) than the patients without TT. The mortality after TT was significantly lower (7% vs. 17%, p=0.015), without fatal outcome in patients with finally successful TT. Reasons against TT application were late arrival to hospital (51%) and contraindications for TT (34%). In patients without TT, the median pain-to-door time and door-to-needle time were significantly longer than in the TT group (7 vs. 2.5 hours and 55 vs. 20 min, respectively; p<0.001). CONCLUSION: Older age, female gender, previous myocardial infarction, and late arrival to the CCU negatively influence the use of TT in AMI. TT should be improved by shortening pain-to-door time, broadening indications, and limiting contraindications. PMID- 11063764 TI - Aortic valve replacement with and without concomitant coronary artery bypass surgery in the elderly: risk factors related to long-term survival. AB - AIM: Preoperative coronary angiography often reveals significant coronary artery lesions in elderly people (>75 years of age) referred to hospital for aortic valve replacement (AVR). However, the possible benefit of concomitant coronary artery bypass grafting (CABG) in elderly is still under debate. In an effort to contribute to this discussion, we evaluated our data on elderly patients after aortic valve replacement. METHODS: Between January 1990 and December 1993, 219 patients, aged 75 years and older, underwent AVR with or without concomitant CABG at our Department. There were 121 patients in the AVR group and 98 patients in the AVR+CABG group. There was no significant difference between the two groups in their age, sex valve type, valve size, and presence of diabetes. Five variables (concomitant CABG, age, sex, and type and size of prosthesis) were investigated with regard to long-term survival assessed by the Kaplan-Meier analysis. Group comparisons of survival were made with the Cox-Mantel log-rank test. RESULTS: Early mortality (<30 days) was 0.8% in the AVR group and 4.1% in the AVR+CABG group. Overall actuarial survival was 77.7+/-4.4% at 52 months. There was significantly longer survival in patients with mechanical valve implant in the AVR group. None of the other 5 investigated variables had a significant influence on the long-term survival. CONCLUSION: Our results suggest that AVR done in elderly is a treatment with excellent surgical results. We could not identify concomitant CABG as a predictor of poor long-term surgical outcome. PMID- 11063765 TI - Association between idiopathic mitral valve prolapse and panic disorder. AB - AIM: To evaluate the association between idiopathic mitral valve prolapse and panic disorder. METHODS: The study comprised 50 patients with idiopathic mitral valve prolapse, 50 patients with panic disorder, and 50 healthy controls. All subjects underwent echocardiographic evaluation. If idiopathic mitral valve prolapse was present, the level of prolapse was measured. To reach psychiatric diagnosis, the Structured Clinical Interview for DSM-IV Axis I Disorders, clinician version, was administered to all subjects. For psychometric evaluation, all subjects completed the Symptom Check List, Beck Depression Inventory, and State and Trait Anxiety Inventory. RESULTS: The prevalence of panic disorder was 16% in patients with idiopathic mitral valve prolapse, and 2% in healthy controls (p=0.03). The average scores on all psychometric scales were the highest in the panic disorder group, whereas average scores in idiopathic mitral valve prolapse group were significantly higher than in the control group (p<0.001). Idiopathic mitral valve prolapse and panic disorder groups displayed similar features of panic attack symptoms. Panic disorder cases with idiopathic mitral valve prolapse and those without it did not significantly differ in terms of psychometric evaluations and clinical symptoms. CONCLUSION: The possibility of comorbidity of panic disorder and idiopathic mitral valve prolapse should be taken into account in the approach to the patients diagnosed with either of the disorders. PMID- 11063766 TI - Renoprotective role of nifedipine during gentamicin therapy: randomized controlled trial. AB - AIM: To investigate the protective effect of nifedipine, a dihydropyridine calcium-channel blocker, on renal function (glomerular and tubular) in patients treated with gentamicin, an aminoglycoside antibiotic. METHODS: Thirty-two patients with gentamicin sensitive upper urinary tract infection have been screened and randomized to two groups. The placebo group was given gentamicin and placebo, and the intervention group gentamicin and nifedipine. Gentamicin was given in slow intravenous injection every 12 hours for 10 days, and nifedipine 10 mg orally, 3 times a day. RESULTS: Nifedipine administration during gentamicin therapy promoted primarily the glomerular filtration. In 62% of the patients treated with nifedipine creatinine clearance increased significantly by the end of the study. In the placebo group, 69% of the patients had a creatinine clearance significantly below the baseline at the end of the study. The decrease in creatinine clearance by more than 50% from the initial values was found in 2 patients (1 in each group). There was a significant increase in gammaGT/creatinine clearance ratio in both groups at the end of therapy, indicating that nifedipine did not prevent the brush-border membranous enzyme release caused by gentamicin. CONCLUSION: Nifedipine has positive effects on renal hemodynamics in patients treated with gentamicin. Most likely, the mechanism of action is an increase in glomerular filtration caused by preglomerular vasodilatation. PMID- 11063767 TI - Intravesically administered ketoprofen in treatment of detrusor instability: cross-over study. AB - AIM: To evaluate the therapeutic efficacy of intravesically administered ketoprofen in patients with urodynamically verified detrusor instability. METHODS: This double-blind randomized placebo-controlled cross-over study included 30 patients with urodynamically verified detrusor instability. Their mean age was 44+/-3.6 years (range 37-49) and the median of the parity was 2 years (1-3). The mean duration of symptoms was 18.3+/-3.1 months (range 14-23). After a 6-week screening, patients were randomized to receive ketoprofen or placebo once a day for 4 weeks. Out of 30 patients, 16 started with ketoprofen, and 14 received placebo. After a week of washing period, 16 patients received placebo, and 14 received ketoprofen. The solution for intravesical application was 50 mL of saline with 2 mL (100 mg) of ketoprofen warmed to 37 degrees C. The placebo solution contained 2 mL of distilled water instead of ketoprofen. The assessment including micturition diaries, cystometric measurements, and bacteriological analysis of urine specimens was performed at the beginning of the study and after the treatment. RESULTS: The subjective cure rate was 18/30 after ketoprofen. The instability index was lower after ketoprofen than before treatment or after placebo (p<0.001). Maximal cystometric capacity and the urinary bladder volume at which the patients felt urgency to void were larger after ketoprofen than before it (p<0.001) or after placebo (p<0.001). The number of patients with uninhibited bladder contractions decreased significantly after ketoprofen, but not after placebo (p<0.001). No side effects were observed. CONCLUSION: Intravesically administrated ketoprofen is a feasible and effective treatment for detrusor instability. PMID- 11063768 TI - Physical activity of urban adult population: questionnaire study. AB - AIM: To assess the level of physical activity of an urban population according to gender, age, smoking, and educational differences. METHODS: The sample comprised 594 men and women living and working in Zagreb, Croatia. Work, sport, and leisure time activity indices were obtained by the Baecke's questionnaire. Significance of differences was tested by the Student's t-test. The relation between the indices and the education was determined by correlation analysis. RESULTS: Women had lower work and sport indices score, and higher leisure-time activity index. After the age of 50, women participated significantly more in sport activities than men. There was a negative correlation between the education and the work index, and a significantly positive correlation between the educational level and the sport activity index, regardless of gender and age. Women smokers participated to a significantly lesser extent in sport activities than women non smokers. Such differences were not observed in men. CONCLUSION: People with lower educational level sustain more workload at their jobs than those with higher education. More educated people participate more in sport activities, although leisure time activity is not significantly related to education. In women, participation in sport activities negatively correlates with the smoking habit. PMID- 11063769 TI - Low sero-prevalence of Lyme borreliosis in the forested mountainous area of Gorski Kotar, Croatia. AB - AIM: Clinical forms of Lyme disease in Gorski Kotar have occurred only sporadically, in contrast to the northwestern Croatia and the neighboring areas of Slovenia, which are well-known Lyme borreliosis endemic regions. Our aim was to assess the level of sero-prevalence of Borrelia burgdorferi sensu lato in a high-risk population of forestry workers in the mountainous region of Gorski Kotar, Croatia, and compare it with the sero-prevalence in the residents of that area and the neighboring littoral region. METHODS: A sero-epidemiological study was conducted on 520 healthy subjects, divided in 3 groups: the first group included 234 forestry workers, residents of Gorski Kotar, the second 100 residents of various professions in the same region, and the third 186 subjects of various professions from the neighboring littoral region. The sera were collected during the winters of two successive years, 1997 and 1998. Lyme borreliosis serology was performed by indirect immunofluorescence assay. Sera from 10 hunting dogs from Gorski Kotar were also analyzed. RESULTS: The IgG antibodies to B. burgdorferi sensu lato were found in 11 examinees (4.7%) from the group of forestry workers, in 3 (3%) from the second group, and in 5 (2.7%) from the third group. Four out of 10 dogs (40%) had IgG antibodies against B. burgdorferi. CONCLUSION: Our results show that the forest and mountainous area of Gorski Kotar, Croatia, has the characteristics of a low sero-prevalence area, in contrast to the endemic neighboring areas. PMID- 11063770 TI - Epidemiology of bronchial asthma among schoolboys in Al-Khobar city, Saudi Arabia: cross-sectional study. AB - AIM: To determine the prevalence of bronchial asthma among Saudi Arabian schoolboys living in Al-Khobar city. METHODS: Parents of 1,482 randomly selected boys in preparatory and elementary schools included in this cross-sectional study filled out a questionnaire on asthma signs in their sons. Number of Questionnaire Diagnosed Asthmatics was compared with the number of Physician-Diagnosed Asthmatics. RESULTS: The prevalence rates of questionnaire-diagnosed asthma (QDA) and physician-diagnosed asthma (PDA) were 9.5% and 8.1%, respectively. Rates of allergic diseases were significantly higher among QDA schoolboys and their parents. They reported to be more exposed to allergenic environmental factors (pets at home, passive smoking) than non-QDA. CONCLUSION: The prevalence of QDA among schoolboys in Al-Khobar city is higher than the prevalence of QDA in other Arabian, developing, or European countries, but still lower than in other parts of Saudi Arabia. A combination of genetic and environmental factors may explain this finding. PMID- 11063771 TI - Post traumatic stress disorder with psychotic features. AB - AIM: To study the combined course and interaction of post-traumatic stress disorder (PTSD) and psychosis through detailed case studies. METHODS: We described 6 case studies of Israeli veterans with PTSD and psychosis, who were referred to our Center for the evaluation of their psychiatric status. RESULTS: All the patients developed PTSD shortly after the exposure to a combat/military trauma. Psychosis appeared after months or even years, sometimes after a trauma related trigger. Psychotic symptoms, such as delusions or auditory hallucinations, were usually paranoid or depressive and related in content to the traumatic experience. CONCLUSION: The combined course of PTSD and psychotic disorder may reflect two distinct disorders, but in some cases it seems justified to make a diagnosis of PTSD with psychotic features. In addition, it seems that in certain conditions, traumatic exposure and/or PTSD may serve as a trigger for psychosis. PMID- 11063772 TI - Erysipelas-like cellulitis with Pasteurella multocida bacteremia after a cat bite. AB - A 73-year-old female patient presented with Pasteurella multocida erysipelas-like cellulitis, bacteremia, and shock. The onset of the disease occurred 24 h after a cat bit her to the right lower leg. Initially, the picture of bacteremia and shock developed, with minimal local cellulitis. Pasteurella multocida grew in blood culture. A combination of amoxicillin and clavulanic acid was therapeutically successful in respect that the signs of bacteremia and shock disappeared. However, extensive erysipelas-like cellulitis developed on the bitten leg within the next 2 days. The disease was efficiently treated with penicillin G combined with netilmicin and administered for 10 days. This report documents the first case of Pasteurella multocida erysipelas-like cellulitis with bacteremia and shock. PMID- 11063773 TI - Management of deep-vein thrombosis. PMID- 11063774 TI - Management of Deep-vein Thrombosis. PMID- 11063775 TI - Banned weapons. PMID- 11063776 TI - Structural genomics: an overview. PMID- 11063777 TI - Methodologies for target selection in structural genomics. AB - As the number of complete genomes that have been sequenced keeps growing, unknown areas of the protein space are revealed and new horizons open up. Most of this information will be fully appreciated only when the structural information about the encoded proteins becomes available. The goal of structural genomics is to direct large-scale efforts of protein structure determination, so as to increase the impact of these efforts. This review focuses on current approaches in structural genomics aimed at selecting representative proteins as targets for structure determination. We will discuss the concept of representative structures/folds, the current methodologies for identifying those proteins, and computational techniques for identifying proteins which are expected to adopt new structural folds. PMID- 11063778 TI - Towards a covering set of protein family profiles. AB - Evolutionary classification leads to an economical description of the protein sequence universe because attributes of function and structure are inherited in protein families. Efficient strategies of functional and structural genomics therefore target one representative from each family. Enumerating all families and establishing family membership consistently based on sequence similarities are nontrivial computational problems. Emerging concepts and caveats of global sequence clustering are reviewed. Explicit multiple alignments coupled with neighbourhood analysis lead to domain segmentation, and hierarchical unification helps to resolve conflicts and validate clusters. Eventually, every part of every sequence will be assigned to a domain family which is uniquely associated with a fold and a molecular function. PMID- 11063779 TI - Structural proteomics: prospects for high throughput sample preparation. PMID- 11063780 TI - An integrated approach to structural genomics. AB - Structural genomics aims at determining a set of protein structures that will represent all domain folds present in the biosphere. These structures can be used as the basis for the homology modelling of the majority of all remaining protein domains or, indeed, proteins. Structural genomics therefore promises to provide a comprehensive structural description of the protein universe. To achieve this, a broad scientific effort is required. The Berlin-based "Protein Structure Factory" (PSF) plans to contribute to this effort by setting up a local infrastructure for the low-cost, high-throughput analysis of soluble human proteins. In close collaboration with the German Human Genome Project (DHGP) protein-coding genes will be expressed in Escherichia coli or yeast. Affinity-tagged proteins will be purified semi-automatically for biophysical characterization and structure analysis by X-ray diffraction methods and NMR spectroscopy. In all steps of the structure analysis process, possibilities for automation, parallelization and standardization will be explored. Major new facilities that are created for the PSF include a robotic station for large-scale protein crystallization, an NMR center and an experimental station for protein crystallography at the synchrotron storage ring BESSY II in Berlin. PMID- 11063781 TI - Structural genomics projects in Japan. PMID- 11063782 TI - Olanzapine augmentation of fluvoxamine-refractory obsessive-compulsive disorder (OCD): a 12-week open trial. AB - A few studies have tried antipsychotic augmentation in obsessive-compulsive disorder (OCD) patients who are non-responders to selective serotonin reuptake inhibitors. The aim of this study was to investigate the efficacy and tolerability of olanzapine addition to fluvoxamine-refractory OCD patients and to assess if a comorbid chronic tic disorder or a concomitant schizotypal personality disorder was associated with response. Twenty-three OCD non responders to a 6-month, open-label trial with fluvoxamine (300 mg/day) entered a 3-month open-label trial of augmentation with olanzapine (5 mg/day). OC symptom change was measured with the Yale-Brown Obsessive-Compulsive Scale (Y-BOCS) and the Clinical Global Impression (CGI) scale. Differences between responders and non-responders were assessed with regard to age, sex, duration of illness, baseline Y-BOCS score, and comorbidity with chronic tic disorders or schizotypal personality disorder. A significant decrease of mean Y-BOCS score between pre- and post-treatment (26. 8+/-3.0 vs. 18.9+/-5.9) was found at endpoint. Ten patients (43.5%) were rated as responders. The most common side effects were mild to moderate weight gain and sedation. In our sample, three patients (13. 04%) had a chronic motor tic disorder, and four (17.39%) had a codiagnosis of schizotypal personality disorder. Concomitant schizotypal personality disorder was the only factor significantly associated with response. It appears that augmentation of olanzapine in fluvoxamine-refractory OCD may be effective in a large number of patients, including those with comorbid schizotypal personality disorder. PMID- 11063783 TI - Auditory sensory memory in schizophrenia: inadequate trace formation? AB - This study explored duration mismatch negativity reductions observed in individuals with schizophrenia, in particular, the relationship to behavioural measures of temporal discrimination and two event-related potential (ERP) components occurring during the first phase of auditory sensory memory. Twenty two patients with a DSM-IV and ICD-10 diagnosis of schizophrenia and 25 healthy comparison volunteers participated in a behavioural and an ERP testing session. Both groups performed equivalently on behavioural estimates of filled interval duration discrimination and gap detection. In contrast, electrophysiological measures revealed a significant reduction in patients' duration mismatch negativity and a significant difference in patients for the pattern of N100 facilitation over short stimulus onset asynchronies. Whilst behavioural results indicate intact temporal processing of filled intervals and equal temporal resolution limits in schizophrenia, both ERP measures indicated differences in auditory processing that may be traced to activity occurring during the first 250 ms. Results highlight the possibility of abnormalities in the process of auditory trace formation and temporal summation in schizophrenia. PMID- 11063784 TI - Cocaine use and the mid-latency auditory evoked responses. AB - To examine the effects of chronic cocaine use on the mid-latency auditory evoked responses (MLAERs), we recorded the evoked responses of 15 cocaine-dependent subjects and 13 age-matched healthy control subjects. Two evoked response paradigms were used: a trains paradigm with four different inter-stimulus intervals (ISIs) and a paired-click paradigm. Our data suggest that cocaine dependent subjects generate smaller P50 components when long ISIs are used with multiple repetitions (in the trains paradigm). In a single repetition paradigm (paired clicks), a significant decrease in the ability to attenuate the N100 component was seen in the cocaine-dependent subjects. PMID- 11063785 TI - Risk for obstetric complications and schizophrenia. AB - The goal of this study was to determine whether cases with schizophrenia or related disorders show a history of obstetric complications significantly more often than control subjects and, if so, whether the enhanced risk of a negative pregnancy outcome also extends to the non-schizophrenic offspring of cases. Data based on the obstetric birth case-notes of patients with diagnosed schizophrenia or related disorders were compared to those of normal 'healthy' control subjects; each case/control pair was individually matched by gender, time and parity of birth, maternal age and marital status. Forty-four case/control pairs born in Padova (Italy) between 1964 and 1978 were assessed for prenatal and perinatal complications, including abnormal gestational age or birthweight. No significant differences were observed between cases and control subjects in the general characteristics of birth; gestational age and birthweight in particular were strictly comparable between cases and control subjects. The schizophrenia spectrum patients (75%) were more likely than control subjects (59%) to have experienced at least one definite obstetric complication: odds ratio=2.07 (95% CI: 0.83-5. 15). Cases also suffered more complications per birth than control subjects (average 2:1). In particular, obstetric complications involving a clear damaging potential were seen significantly more often among cases than control subjects: 34% vs. 9%, Fisher's exact test, P=0.008 (odds ratio=5.17, 95% CI: 1.55 17.21). Moreover, severe obstetric complications were noted more often among males (n=13, 41%) than females (n=2, 15%). When any previous pregnancies of the mothers of patients were compared with those of the mothers of control subjects, mothers of cases were seen to have suffered unfavorable pregnancy outcomes significantly more often. In particular mothers of cases were seen to have had more miscarriages (OR=4.66), and pre-term births (OR=2.58) than control subects' mothers. Severe, brain-damaging obstetric complications would seem to be a possible antecedent to a diagnosis of schizophrenia or a related disorder in adulthood. Indeed, some early onset cases may be accounted for by prenatal brain lesions. This enhanced risk of negative pregnancy outcome may be under genetic control, contributing to the persistence of schizophrenia in the general population. The 'healthy' status of control subjects was ascertained indirectly, not by individual assessment of the subjects. The sample size limits the statistical power of calculations. PMID- 11063786 TI - Expressed emotion and psychoeducational intervention for relatives of patients with schizophrenia: a randomized controlled study in Japan. AB - The benefit of single-family treatment (SFT) in addition to short educational sessions (SES) consisting of multiple-family treatment was investigated. The study design was a randomized controlled study. Subjects were 30 patients suffering from schizophrenia with at least one of their family members showing high expressed emotion (EE) in the Camberwell Family Interview. After the SES, the patients were randomly allocated to two groups: those who received routine individual outpatient treatment and those who received additional SFT and routine treatment. The two groups were followed for 9 months after discharge, and the relapse risks were compared. The relapse risk was lower in the SES+SFT group than in the SES group (23.1% vs. 35.3%). However, the difference was not significant. When high-EE families were classified into those with many critical comments (high-CC) or a high score of emotional overinvolvement (high-EOI), the relapse risk was 0% in the patients living with a high-CC family not only in the SES+SFT group but also in SES group. In the patients living with a high-EOI family, the relapse risk was lower in the SES+SFT group than in the SES group (42.9% vs. 60.0%). These findings suggest that high-EE families should receive at least SES, and additional SFT should be given to families with specific needs. PMID- 11063787 TI - Temporalis exteroceptive suppression in generalized anxiety disorder and major depression. AB - The duration abnormality of the exteroceptive suppressions, or silent periods, of peripheral or jaw-closing muscle activities induced by transcranial magnetic or trigeminal electrical stimuli in patients suffering from anxiety or depression is ill-defined. We therefore studied the exteroceptive suppression periods of the temporalis muscle electromyography elicited by trigeminal territory electrical stimuli, Zuckerman-Kuhlman's Personality Questionnaire, and Plutchik-van Praag's Depression Inventory (PVP) in 12 patients suffering from generalized anxiety disorder (GAD) and 16 from major depression (MD) as well as in 17 healthy volunteers. Durations of the second suppression period (ES2) sociability scores were decreased in GAD patients, while PVP and neuroticism-anxiety scores were elevated in both GAD and MD patients. There was a positive correlation between ES2 duration and sociability score in the GAD group. This study indicates that anxiety can modify the temporalis ES2 duration through cortical descending inhibitory controls. PMID- 11063788 TI - No evident neuronal damage after electroconvulsive therapy. AB - Electroconvulsive therapy (ECT) is regarded as one of the most effective treatments for major depressive disorder but has also been associated with cognitive deficits possibly reflecting brain damage. The aim of this study was therefore to evaluate whether ECT induces cerebral damage as reflected by different biochemical measures. The concentrations in the cerebrospinal fluid (CSF) of three established markers of neuronal/glial degeneration, tau protein (tau), neurofilament (NFL) and S-100 beta protein, were determined in nine patients who fulfilled DSM-IV criteria for major depression. CSF samples were collected before and after a course of six ECT sessions. The CSF/serum (S) albumin ratio reflecting potential blood-brain barrier (BBB) dysfunction was also determined at these time points. The treatment was clinically successful with a significant decline of depressive symptoms in all patients as assessed by the Montgomery-Asberg Rating Scale for Depression. Several patients had signs of BBB dysfunction and/or neuronal damage before the start of treatment. Levels of CSF tau, CSF-NFL and CSF-S-100 beta levels were not significantly changed by ECT. Also the CSF/S albumin ratio was found to be unchanged after the course of ECT. In conclusion, no biochemical evidence of neuronal/glial damage or BBB dysfunction could be demonstrated following a therapeutic course of ECT. PMID- 11063789 TI - Relationship between serum cholesterol levels and meta-chlorophenylpiperazine induced cortisol responses in healthy men and women. AB - We investigated the effect of cholesterol on serotonergic receptor function in 20 healthy male and 10 healthy female subjects using cortisol responses to meta chlorophenylpiperazine (m-CPP) neuroendocrine challenge tests. M-CPP, a metabolite of the antidepressant trazodone, has been widely used in psychopharmacology research as a probe of serotonin function. In the human brain, m-CPP binds both to various serotonergic receptors, mainly 5-HT(2C), and to alpha(2)-adrenoceptors. After an overnight fast, the subjects received m-CPP (0.5 mg/kg) or identical placebo capsules orally in a randomized, double blind, crossover design. Blood was obtained for measurement of cholesterol and cortisol. In some analyses, especially in males, there were significant positive correlations between serum cholesterol levels and cortisol responses. These findings suggest the possibility that serum cholesterol levels may be positively associated with serotonergic receptor function. The existence of such an association may provide an explanation for reported increases in depression, suicide and violence in individuals with low or lowered cholesterol. PMID- 11063790 TI - Serum cholesterol levels in paranoid and non-paranoid schizophrenia associated with physical violence or suicide attempts in Taiwanese. AB - A review of medical charts over a 1-year period was carried out in a population of 213 Taiwanese psychiatric inpatients that included 106 patients with schizophrenia. In subgroup analyses within the group of patients with schizophrenia, no significant differences in serum cholesterol levels were found between paranoid and non-paranoid schizophrenic patients, between patients with and without physical violence, or between patients who had and had not made a suicide attempt. PMID- 11063791 TI - Association study of schizophrenia spectrum disorders and dopamine D3 receptor gene: is schizoaffective disorder special? AB - Alterations in dopamine neurotransmission have been hypothesized to play a role in the etiology of schizophrenia. We considered the dopamine D3 receptor gene on chromosome 3 as a candidate gene for an association analysis. We compared PCR based genotype markers for healthy controls (n=120) and patients (n=95) with schizophrenia and schizophrenia spectrum disorders as diagnosed by consensus according to DSM-III-R. Our results possibly indicate an association of schizoaffective disorder with DRD3 homozygosity (P=0.056). PMID- 11063792 TI - Genetic characterization of angiomatoid fibrous histiocytoma identifies fusion of the FUS and ATF-1 genes induced by a chromosomal translocation involving bands 12q13 and 16p11. AB - This case report documents the first karyotypic, fluorescence in situ hybridization, and genetic analysis of an angiomatoid fibrous histiocytoma that arose and recurred in the arm of a 5.5-year-old girl. Complex rearrangements between chromosomes 2, 12, 16, and 17 were noted, as well as deletion in the long arm of chromosome 11. Flow cytometry revealed a normal cell population. The t(12;16) site was further investigated using reverse transcriptase-polymerase chain reaction. We found that the FUS (also known as TLS) gene from 16p11 combined with the ATF-1 gene from 12q13 to generate a chimeric FUS/ATF-1. The FUS gene is rearranged in the t(12;16)(q13;p11) that characterizes myxoid liposarcoma and in acute myeloid leukemia with t(16;21)(p11;q22), while the ATF-1 gene is rearranged in the t(12;22)(q13;q12) found recurrently in clear cell sarcomas (malignant melanoma of soft parts). Thus, the FUS/ATF-1 gene in angiomatoid fibrous histiocytoma is predicted to code for a protein that is very similar to the chimeric EWS/ATF-1 found in clear cell sarcoma. PMID- 11063793 TI - Establishment of a human malignant fibrous histiocytoma cell line, COMA. Characterization By conventional cytogenetics, comparative genomic hybridization, and multiplex fluorescence In situ hybridization. AB - The human COMA cell line has been established from a storiform pleomorphic malignant fibrous histiocytoma (MFH). As expected for this tumor type, a very complex karyotype was observed after R-banding analysis. An extensive analysis by 24-color painting, comparative genomic hybridization (CGH), and fluorescence in situ hybridization (FISH) was performed. Twelve complex marker chromosomes recurrently observed were clearly identified; among them, three were systematically present in all analyzed metaphases. Amplifications detected by CGH were refined by FISH with probes specific for various candidate loci. A significant aneuploidy and numerous micronuclei were observed, which could be related to the anomalies of centriole numbers detected in a proportion of cells. Such an analysis, performed on a series of MFH cell lines, would allow the delineation of the genomic alterations specific for the oncogenesis or progression of this complex tumor type or both. PMID- 11063794 TI - Comparative genomic hybridization reveals recurrent enhancements on chromosome 20 and in one case combined amplification sites on 15q24q26 and 20p11p12 in glioblastomas. AB - We examined homogenized tissue samples of biopsies from 19 astrocytomas of different grades for genetic imbalances using comparative genomic hybridization (CGH): three astrocytomas grade II, and 16 astrocytomas grade IV (glioblastoma multiforme), one of the glioblastomas representing the recurrence of a benign oligoastrocytoma. In two of three cases of astrocytoma grade II, a gain of chromosome 7 was found. The alterations in the glioblastomas were complex, and most frequently showed the characteristic gain of chromosome 7 and loss of chromosome 10. The single analyzed case of recurrence of an oligoastrocytoma was characterized by a unique CGH pattern. This tumor showed two distinct alterations: apart from an amplification on 15q24q26, we found a distinct amplification of a small region on 20p11.2p12, which has not been previously described in brain tumors. Partial or complete gains of chromosome 20 arose in six other tumors; we conclude that chromosome 20 in particular 20p11. 2p12, may harbor relevant genes for glioma progression. PMID- 11063795 TI - Unusual breakpoint distribution of 8p abnormalities in T-prolymphocytic leukemia: a study with YACS mapping to 8p11-p12. AB - Chromosome 8 abnormalities are seen in 80% of patients with T-cell prolymphocytic leukemia (T-PLL). The abnormalities described are idic(8)(p11),t(8;8)(p11;q12),+8, and 8p+ with the involvement of 8p. To localize 8p11-p12 breakpoints in T-PLL, metaphases from seven cases were karyotyped. Those with idic(8)(p11) and add(8)(p11) were probed with a panel of contiguous YACs derived from 8p11-p12 using fluorescence in situ hybridization (FISH). Analysis of FISH results showed that 8p11-p12 breakpoints cluster into two regions. The first region is telomeric to YAC 899e2, which contains the fibroblast growth factor receptor-1 gene (FGFR1) and appears to cluster within a 1.5-MB YAC 807a2. The second region is more centromeric with breakpoints on either side of YAC 806e9, flanked by YAC 940f10 distally and YAC 910d7 proximally, the latter containing the MOZ gene. These findings showed that a segment of 8p was still present in the isodicentric, but the pattern of clustering does not seem to correspond to a breakpoint affecting a single gene. The clustering regions are likely to be hot spots for recombination and result in idic(8)(p11) and 8p+. These changes point to the pathogenesis of T-PLL involving deletion of a gene sequence on 8p and/or gain of a copy of 8q. PMID- 11063796 TI - Reciprocal translocation (3;5)(q26;q22) and possible BCHE gene involvement in an unusual myelogenous disorder with both myeloproliferative and dysplastic features. AB - We report on a 77-year-old male patient who presented with an unusual myelogenous disorder exhibiting both myeloproliferative and dysplastic features. The patient suffered from leukocytosis, eosinophilia, basophilia, transfusion dependent anemia, and rapidly progressing thrombocytopenia. Classical chromosome analysis and fluorescence in situ hybridization (FISH) revealed a reciprocal t(3;5)(q26;q22). Using yeast artificial chromosome (YAC) probes, the breakpoint on chromosome 3 was localized to the butyrylcholinesterase (BCHE) gene (3q26.1 q26.2). This gene has recently been implicated in the regulation of myeloid cells. Whether the BCHE gene was also involved in the deregulation of myelopoiesis, causing the unusual clinical picture in this case, remains unknown. PMID- 11063797 TI - Concomitant amplification and expression of PAX7-FKHR and MYCN in a human rhabdomyosarcoma cell line carrying a cryptic t(1;13)(p36;q14). AB - Alveolar rhabdomyosarcoma (ARMS) is associated with the specific chromosomal translocation (2;13)(q35;q14) or its rarer variant t(1;13)(p36;q14), which produces the fusion gene PAX7-FKHR. Here we describe the human cell line RC2, derived from an ARMS, which harbors a cryptic t(1;13)(p36;q14) and concomitantly shows amplification of the PAX7-FKHR fusion gene and of the MYCN oncogene. The t(1;13) and MYCN oncogene were studied by standard cytogenetic analysis and molecular techniques. The reverse transcriptase polymerase chain reaction demonstrated the expression of PAX7-FKHR mRNA in RC2 cells, although karyotype analysis failed to demonstrate a t(1;13)(p36;q14) chromosomal translocation or a derivative 13 chromosome. Double minute chromosomes were detected in all the metaphases studied. Fluorescence in situ hybridization analysis revealed multiple copies of the PAX7-FKHR fusion gene localized exclusively on a subset of double minutes, whereas multiple copies of MYCN were identified on other double minute chromosomes. Southern-blot analysis demonstrated that RC2 cells contain approximately 20 copies of the MYCN oncogene. So far no continuous RMS cell line carrying the t(1;13)(p36;q14) has been described, and PAX7-FKHR and MYCN amplifications have always been reported to occur separately in rhabdomyosarcoma (RMS). The availability of an ARMS cell line that harbors the t(1;13)(p36;q14) constitutes a useful tool for further understanding the role of the PAX7-FKHR fusion gene in RMS oncogenesis and may improve knowledge of the possible relation between PAX7-FKHR and MYCN amplification. PMID- 11063798 TI - Molecular cytogenetic study of a hemangiopericytoma in a newborn. AB - Two balanced reciprocal chromosome translocations, t(8;12)(p21;p13. 1) and t(15;16)(q24;q22), characterized a rare hemangiopericytoma in a newborn. Chromosome painting with a chromosome microdissection-derived whole-chromosome 8 probe confirmed that the t(8;12) was due to a reciprocal translocation. To the best of our knowledge, these chromosome findings are unique to this unusual case of a pediatric hemangiopericytoma. PMID- 11063799 TI - p53 abnormality and chromosomal instability in the same breast tumor cells. AB - To clarify the important role of the tumor-suppressor gene p53 in maintaining genetic integrity, we estimated chromosome instability and staining of overexpressed p53 protein in the same cells of five primary breast carcinomas. The method included both fluorescence immunohistochemistry and fluorescence in situ hybridization (FISH) on sections from formalin-fixed, paraffin-embedded breast cancer tissue. By using a centromeric FISH probe for chromosome 17 on interphase cells in these sections, we showed that cells with abnormal p53 protein expression had a statistically significant higher number of chromosome 17 than did cells with no p53 protein staining in the same samples as well as cells in four other tumor samples with no p53 protein staining. The samples identified positive for p53 abnormality by immunostaining were shown to have p53 mutation by constant denaturing gel electrophoresis analysis and DNA sequencing. These mutated samples were characterized by high DNA index, high S-phase, abnormal karyotype, and aneuploidy. The results strongly implicate p53 mutation as a cause for chromosomal instability and a crucial step in mammary carcinogenesis. PMID- 11063800 TI - Losses of heterozygosity in endometrial adenocarcinomas: positive correlations with histopathological parameters. AB - We analyzed 37 samples of endometrial adenocarcinoma for loss of heterozygosity (LOH) by using a panel of 44 microsatellites located in 29 chromosomal regions. The aim of our study was to investigate the existence of a possible preferential involvement of some tumor suppressor genes in endometrial carcinogenesis. The analysis was performed on tumoral tissue and on a corresponding normal tissue by the use of polymerase chain reaction (PCR) and the comparison of the amplified alleles. We observed significative LOH (>20%) in the chromosomal regions of 2q14 (33.33%), 7q35 (24.00%), 10q22.1 (37. 50%), 11q13-q14 (44.12%), 15q26 (40.63%), 17p13 (25.71%), and 17q21. 3 (37.04%). We defined a 1-cM minimal common deletion in 11q13-q14 between D11S911 and D11S937 markers. A statistical analysis revealed a positive correlation between LOH of 11q13-q14 and clinicopathological data. PMID- 11063801 TI - Cytogenetic and fluorescence in situ hybridization studies in four cases of plasma cell leukemia. AB - We present a cytogenetic and fluorescence in situ hybridization (FISH) study, using centromeric probes for chromosomes 3, 7, 11, and 18, TP53 gene (17p13), and RB-1 locus (13q14) DNA probes, in four cases of plasma cell leukemia (PCL). Among the four cases, three presented monosomy of the RB-1 locus and one monoallelic deletion of the TP53 gene. The present report shows the usefulness of the FISH technique to detect abnormalities not previously observed by conventional cytogenetics. PMID- 11063802 TI - Isochromosome (7)(q10) in Shwachman syndrome without MDS/AML and role of chromosome 7 anomalies in myeloproliferative disorders. AB - Shwachman syndrome (SS) is an autosomal recessive disorder in which bone marrow dysfunction is observed, with development of myelodysplastic syndromes (MDS) and acute myeloid leukemias (AML) in up to one third of the cases. Inconclusive data are available as to increased chromosome breakage in SS, while chromosome 7 anomalies, and often an isochromosome (7)(q10), are frequent in cases with MDS/AML. We report on the consistent presence of an i(7)(q10) in the bone marrow and blood lymphocytes in one of two sisters affected with SS without any clinical or cytological signs of MDS/AML. Thus, this patient was either a case of constitutional mosaicism for the i(7)(q10), or this had to be acquired in a nondysplastic and non-neoplastic marrow clone. DNA polymorphism analysis demonstrated the paternal origin of the i(7q). We postulate that the SS mutation acts as a mutator gene, and causes karyotype instability; abnormal clones would thus arise in the marrow, and chromosome 7 anomalies, i(7q) in particular, will in turn lead to MDS/AML. If this interpretation is correct, it would be also an indication to consider chromosome 7 anomalies in general, out of SS, as primary changes in MDS/AML pathogenesis. PMID- 11063803 TI - A novel high mobility group protein gene is a candidate for Xp22 abnormalities in uterine leiomyomas and other benign tumors. AB - Because genes of the high mobility group protein family HMGI(Y) are known to take part in the development of a variety of benign solid tumors, the aim of the present study was to search for further members of that family in the human genome. Analysis for HMGI(Y)-related sequences by the polymerase chain reaction (PCR) with the use of cDNA-specific primers offered evidence for HMGIY-like sequences, whereas HMGIC-related sequences were apparently absent. By chromosomal assignment of somatic cell hybrids PCR, HMGIY cDNA-related sequences were detected on seven chromosomes. Positive clones were obtained by screening of a P1 derived artificial chromosome library and mapped by fluorescence in situ hybridization. One of these clones assigned to Xp22.1 was chosen for further analysis because Xp22 is a target region for clonal aberrations in benign solid tumors. Sequence analysis of a DNA fragment of this clone, designated as HMGIYL1, revealed a 94.4% homology to the coding region of HMGIY. Within the HMGIYL1 sequence, no nucleotide sequence divergences leading to a frame shift or a new termination codon compared to HMGIY were found, and a TATA-box-like motif 5' of it was detected. By reverse transcriptase PCR experiments with the use of HeLa cells and human fetal tissue, HMGIYL1 expression was not detectable. Nevertheless, if not active by itself, it is possible that HMGIYL1 may become activated by chromosomal rearrangements of Xp22 observed in benign solid tumors. PMID- 11063804 TI - Mutational analysis of the NM23.H1 gene in human breast cancer. AB - NM23.H1 is a protein connected with tumor progression. Loss of heterozygosity and reduced expression of the gene have been associated with poor prognosis and increased incidence of metastases in many epithelial tumors. The aim of this study was to detect the presence of NM23.H1 point mutations or small deletions in human breast carcinomas by using the single-strand-conformation polymorphism (SSCP) technique. Mutational analysis was performed on 76 breast tumors, 10 of which had allelic deletion of the gene. The NM23.H1 mRNA content also was evaluated in each sample. Only a C-to-A transversion leading to a stop codon was found in the 5' untranslated region of exon 1. A polymorphic SSCP pattern was identified in exon 1; direct sequencing showed a C-to-T transition 30 nucleotides upstream from the 5' splice site flanking exon 1. None of the tumors analyzed presented both alleles inactivated. Our results suggest that NM23.H1 is rarely inactivated by point mutations. PMID- 11063805 TI - Trisomy 15 is frequently observed as a minor clone in patients with Anemia/MDS/NHL and as a major clone in patients with AML. AB - Trisomy 15 as the sole karyotypic aberration is an uncommon clonal cytogenetic aberration in hematological malignancies, making its significance unclear. Previous studies have reported relations of trisomy 15 with low-grade myelodysplasia or a benign age-related phenomenon associated with loss of the Y chromosome. To define the significance of trisomy 15, we conducted a retrospective study of all examples of trisomy 15 accessed in our laboratories. Trisomy 15 was observed as a clonal abnormality (> or =2 cells) in 17 cases and nonclonal (single cell) in 9 cases. The majority of cases (14/17 clonal cases) had a minor clone (5-35% of metaphase cells) of trisomy 15. The minority of cases (3/17) had a major clone (80-95% of metaphase cells) of trisomy 15. Two of these 3 cases were diagnosed as having acute myelocytic leukemia. Fluorescence in-situ hybridization (FISH) with the use of a chromosome 15-specific alpha-satellite probe was performed on 3 of 17 clonal cases and on 3 of 9 nonclonal cases. FISH results revealed the presence of a minor clone (from 3 to 5 of 700 interphase cells) in 5 of them, 2 of which had trisomy 15 in 20% of metaphase cells. These results may indicate that the 20% of trisomy 15 are very likely an overrepresentation of a very minor clone that could be transitory. In summary, the analysis of our cytogenetic and FISH results revealed the presence of two types of trisomy 15 clones: a minor clone that could be transitory or indolent and a major clone that could be of a neoplastic nature. PMID- 11063806 TI - Karyotypic findings in two cases of male breast cancer. AB - Male breast cancer is uncommon; so far, only 10 cases with chromosome banding analysis have been published. We report the cytogenetic findings of two invasive breast cancers in two Caucasian men lacking a history of familial breast cancer and more than 70 years of age. Both had ductal carcinomas with lymphangiosis carcinomatosa and positive lymph nodes at diagnosis. Strong expression of estrogen receptor, weak expression of progesterone receptor, and lack of expression of androgen receptor by both tumors were demonstrated by immunohistochemistry, as well as lack of expression of p53 and C-ERB-B-2. The karyotypes were 45 approximately 46,XY,-Y[4],-7[2],+8[2],t(8;12)(q21;q24)[3], del(9)(q22)[3],del(11)(p11p14)[5],del(18)(q21)[7], t(19;20)(p10;q10)[8] [cp13] and 61 approximately 69,XXXY,-Y[3], del(2)(p21)[4],del(3)(p22q26)[3],-4, 4[5],+5,+5[5], dic(5;11)(p14;q23)[3],del(6)(q23)[4],del(8)(p21)[3],-9[4],-11[4],+ i(12)(p10)[4],-16[3],del(17)([13)[5],del(18)(q21)[4],+19[5], +20[4][cp7], respectively. Although the available data on male breast cancer are still very limited, our findings confirm that gain of an X chromosome, loss of the Y chromosome, gain of chromosome 5, and loss of material from chromosomes 17 and 18 are nonrandom aberrations in male breast cancer. Trisomy 8, characteristic of ductal carcinomas, was found in one case. PMID- 11063807 TI - Dicentric (17;18) in a case of atypical B-cell chronic lymphocytic leukemia. AB - We report a new dic(17;18)(p11.2;p11.2) in a 61-year-old male patient diagnosed with atypical B-cell chronic lymphocytic leukemia. The dic(17;18)(p11.2;p11.2) was detected in 90%, 10%, and 100% of metaphases in the peripheral blood, bone marrow, and lymph node, respectively. Fluorescence in situ hybridization studies with chromosome 17 and 18 centromeric probes revealed the presence of two normal centromeres of both chromosomes 17 and 18. The centromere of one chromosome 17 was found together with the centromere of one chromosome 18, confirming the dicentric nature of the rearrangement. In addition, with the use of a 17p13.1 region probe, monosomy of the 17p13 region, where the Tp53 gene is located, was observed. PMID- 11063808 TI - Amplification of the MLL region in acute myeloid leukemia. AB - We report amplification of the MLL gene region (11q23-->11qter) in a 72-year-old woman with myelodysplastic syndrome progressing to acute myelomonocytic leukemia and in a 51-year-old man with a history of hairy cell leukemia and secondary myelodysplasia progressing to acute myelogenous leukemia. The amplicons containing MLL were shown by molecular cytogenetics to extend from chromosomal region 11q23 to the distal long arm of chromosome 11 and to be present in the first patient in five copies on a large ring chromosome and present in the second patient also in five copies on two derived chromosomes. Other karyotypic findings in the first patient included del(5q), +8, and der(21)t(17;21), resulting in the loss of a copy of 17p, whereas deletion 7q was observed in the second patient. Southern-blot analysis for the second patient was consistent with MLL amplification but did not demonstrate rearrangement of the germ-line MLL band. Amplification of MLL and the 11q23 region has been documented in only a few cases and appears to be yet another mechanism by which MLL contributes to the leukemia phenotype. PMID- 11063809 TI - Inv(12)(q15q24): a nonrandom change associated with myelodysplasia? AB - A patient with refractory anemia and a paracentric inversion of chromosome 12, inv(12)(q15q24), is described. This is the second reported case with this chromosome anomaly, suggesting that this rearrangement is a rare but nonrandom change associated with myelodysplastic syndromes. PMID- 11063810 TI - A novel dicentric deleted chromosome 21 arising from tandem translocation. AB - We present a 26-year-old patient with myelodysplastic syndrome (MDS). Initial bone marrow cytogenetics with G-banding showed a rearranged chromosome 21, which was dicentric and bisatellited on CBG- and NOR-banding. Fluorescence in situ hybridization helped to characterize the structure, using a whole chromosome 21 paint and the locus specific AML1 gene probe. The rearranged 21 consisted solely of chromosome 21 material, contained only one copy of AML1, and was not a trisomy, but a deleted tandem translocation. The MDS transformed to acute myeloid leukemia (AML), and the patient died almost 12 months post-diagnosis. Cytogenetics was performed three times during the course of the disease, and the dicentric chromosome 21 was present throughout. Although there are a number of published rearrangements of chromosome 21 in MDS and AML, most are isodicentrics. We could not find another case of an abnormal chromosome 21 with the same structure as reported here. PMID- 11063811 TI - Trisomy 4 and double minutes in acute myeloid leukemia: further evidence that double minutes can occur as the primary cytogenetic abnormality. AB - The specific association of trisomy 4 and double minutes (dmin) is rare and is usually reported in patients with acute myeloid leukemia (AML), primarily M2 and M4 subtypes. Several previous reports describing this combination suggested that trisomy 4 was the primary cytogenetic abnormality, and that the presence of the dmin was secondary. We describe a 79-year-old male who presented with myelodysplasia, transforming to AML-M2. Cytogenetic analysis of bone marrow aspirate cultures showed a 46,XY,dmin[12]/47,XY,+4,dmin[7]/46, XY[6] karyotype. The number of dmin ranged from 1 to 150. Fluorescence in situ hybridization (FISH) analysis showed that the dmin were derived from amplification of the MYC oncogene. Dual-color interphase FISH analysis was performed with D4Z1 and MYC probes and showed no evidence of a clone containing trisomy 4 without dmin. These data suggest that dmin may also occur as the primary cytogenetic abnormality in patients with trisomy 4 and dmin. PMID- 11063812 TI - Deletion of 3'-CBFB gene in association with an inversion (16)(p13q22) and a loss of the Y chromosome in a 2-year-Old child with acute myelogenous leukemia-M4. AB - Inversion 16(p13q22) is most commonly associated with acute myelomonocytic leukemia with abnormal eosinophils (M4). In association with this inversion, a proximal deletion at the 16p13 primary arm breakpoint occurs in 20% of cases. We report on a first case of inversion 16 with a distal deletion at the primary arm breakpoint 16q22, detected by using the fluorescent-labeled dual-color probe CBFB. PMID- 11063813 TI - Pentasomy 8 in pediatric myelodysplastic syndrome. AB - Myelodysplastic syndrome developed in a 2-year-old child with the karyotype revealing the presence of pentasomy 8. The significance of the coexistence of pentasomy 8, tetrasomy 8, and trisomy 8 detected by fluorescence in situ hybridization in his marrow is discussed. PMID- 11063814 TI - Fluorescence in situ hybridization determination of 22q12-q13 deletion in two intracerebral ependymomas. AB - The sole cytogenetic abnormalities encountered in two childhood anaplastic intracerebral ependymomas were an isodicentric chromosome 22 in one case and an unbalanced chromosome 22 translocation associated with a partial deletion in the other. Fluorescence in situ hybridization analysis showed that the common 22q arm loss did not involve the rhabdoid region but included the EWS and NF2 loci. These results, in conjunction with data in the literature, suggest that the most frequently recurrent genomic loss in ependymomas does not involve the proximal 22q11.2 chromosome region but is localized distally to the hSNF5/INI1 locus. A tumor-suppressor gene, independent of the NF2 gene, which seems to be exclusively involved in intramedullary spinal cord ependymomas, might be implicated in the genesis of these intracranial tumors. PMID- 11063815 TI - Involvement of interleukin-1 in glial responses to lipopolysaccharide: endogenous versus exogenous interleukin-1 actions. AB - Interleukin-1beta (IL-1beta) participates in neuroinflammation and neurodegeneration. Its mechanisms of action are not fully understood, but appear to involve complex interactions between neurons and glia. The objective of this study was to determine the involvement of endogenous IL-1beta in inflammatory responses to LPS in cultured mouse glial cells, and compare this to the effects of exogenous IL-1beta. Activation of primary mixed glial cultures by incubation with LPS (1 microgram/ml, 24 h), caused marked (approximately ten-fold) increases in release of NO, twenty-fold increases in PGE(2) and ninety-fold increases of IL 6 release. Incubation with human recombinant IL-1beta (100 ng/ml) also stimulated NO and IL-6 release to a similar extent to LPS, but IL-1beta (1 or 100 ng/ml) caused only modest increases (approximately seven-fold) in PGE(2) release. Co incubation with IL-1ra inhibited the effects of LPS on NO release (-65%) and IL-6 production (-30%), but failed to reduce PGE(2) release. These results indicate that exogenous IL-1beta induces release of NO, PGE(2) and IL-6 in mixed glial cultures, and that endogenous IL-1beta mediates inflammatory actions of LPS on NO and to a lesser extent IL-6, but not on PGE(2) release in mixed glial cultures. Indeed endogenous IL-1beta appears to inhibit LPS-induced PGE(2) release. PMID- 11063816 TI - Cryptococcal glucuronoxylomannan delays translocation of leukocytes across the blood-brain barrier in an animal model of acute bacterial meningitis. AB - In bacterial meningitis, neurological damage is associated with a high influx of polymorphonuclear leukocytes (PMN) into the brain. Previous data suggest that the capsular component of the fungus C. neoformans, glucuronoxylomannan (GXM), interferes with PMN-migration into the cerebrospinal fluid (CSF). Therefore, a rabbit model of bacterial meningitis was treated intravenously with GXM. This resulted in (1) a reduction of PMN in the CSF at 6 h (P=0.05), (2) reduced peak TNF-alpha concentrations in the CSF, and (3) diminished tissue inflammation and intravascular margination of PMN in GXM-treated animals. Thus, GXM may represent a novel adjuvant anti-inflammatory agent in bacterial meningitis. PMID- 11063817 TI - A role for adrenoceptors in the regulation of pleural neutrophilia induced by LPS. AB - The role of catecholamines in regulating pleural neutrophilia evoked by intrathoracic (i.t.) injection of lipopolysaccharide (LPS) was investigated in Wistar rats by means of surgical adrenalectomy, depletion of catecholamine stores or adrenoceptor blockade. Treatment of animals with a single dose of LPS evoked a dramatic increase in the number of pleural neutrophils concomitant with an increase in the number of these cells in blood at 4 h. Although blood neutrophilia was drastically reduced when catecholamine stores were depleted with intraperitoneal (i.p.) injection of reserpine, pleural neutrophilia was not modified. However, the i.t. injection of reserpine reduced the increase in pleural neutrophils after LPS stimulation. Adrenalectomy failed to inhibit the increase in neutrophil counts in the blood or pleural cavity after LPS challenge. Pretreatment with intravenous (i.v.) injection of prazosin, an alpha(1)/alpha(2B) antagonist, reduced LPS-induced blood but not pleural neutrophilia. On the other hand, although pleural neutrophilia was not affected by systemic pretreatment with the alpha(2)-adrenoceptor antagonist, yohimbine, the local treatment (i. t. injection) with this antagonist markedly reduced the increase in pleural neutrophil counts observed after stimulation by LPS. In contrast, pleural neutrophilia induced by i.t injection of formyl-methionyl-leucyl-phenylalanine (fMLP) was not modified by local treatment with yohimbine. Taken together, our results suggest that catecholamines, through activation of alpha(1) and alpha(2) adrenoceptors, play a role in the regulation of blood and pleural neutrophilia observed during the inflammatory response evoked by LPS in the pleural cavity. PMID- 11063818 TI - Screening of several H-2 congenic mouse strains identified H-2(q) mice as highly susceptible to MOG-induced EAE with minimal adjuvant requirement. AB - We identified H-2(q) as a susceptible genotype for MOG-induced EAE by systematic screening of a series of H-2 congenic B10 mouse strains. A series of H-2(q) bearing strains with divergent gene backgrounds were subsequently investigated. DBA/1 mice were highly susceptible to MOG(1-125)- and MOG(79-96)-induced EAE in the absence of pertussis toxin. Immunisation with MOG(1-125) and MOG(79-96) induced an autoreactive T-cell response in DBA/1 mice. Brain histopathology revealed T-cell and macrophage-infiltrated lesions with associated demyelination. The important features which make this an appropriate model of human disease are high sensitivity to MOG and dependence of an immunodominant peptide region homologous to that implicated in multiple sclerosis. PMID- 11063819 TI - Naked DNA vaccination differentially modulates autoimmune responses in experimental autoimmune encephalomyelitis. AB - Vaccination with naked DNA represents a therapeutic strategy currently under consideration in multiple sclerosis (MS). In this study, we tested the potential therapeutic effect of vaccination with a naked DNA construct encoding proteolipid protein (pRc/CMV-PLP) upon the outcome of subsequent sensitization for experimental autoimmune encephalomyelitis (EAE) actively-induced in SJL mice with PLP139-151 peptide in adjuvant. Intramuscular vaccination with the naked DNA pRc/CMV-PLP construct led to PLP expression in local muscle tissue that persisted for about 8 weeks. Early sensitization for EAE (4 weeks after DNA vaccination) caused recipient mice to develop a severe, exacerbated form of disease (in comparison to control mice), while late sensitization (>10 weeks) resulted in a milder, ameliorated form. In the groups sensitized <10 weeks post-DNA vaccination with pRc/CMV-PLP induction of a Th1-type cytokine response was noted. In contrast, sensitization >10 weeks post-DNA vaccination led to peripheral tolerance as evidenced by a decrease in T cell proliferation and cytotoxic T cell response, no Th2 response, and no increase in apoptosis. These data are novel in that they demonstrate a differential effect of DNA vaccination and have important implications for its use as a mechanism to enhance or modulate immune reactivity. PMID- 11063820 TI - Ischemic-like condition releases norepinephrine and purines from different sources in superfused rat spleen strips. AB - Transmitters and cotransmitters of the sympathetic nervous system are involved in the regulation of a variety of immune cell functions. However, it is not entirely clear what stimuli lead to the release of these molecules in immune organs. In this study, we investigated whether local ischemia can cause the parallel release of norepinephrine and its cotransmitter, ATP, in the spleen. Ischemic-like conditions, simulated by transient (15 min) O(2) and glucose deprivation, elicited a reversible increase in the release of both norepinephrine and purines from superfused spleen strips preloaded with [3H]norepinephrine or [3H]adenosine. HPLC analysis of the released tritium label revealed a net increase in the amount of ATP, ADP, AMP, adenosine, inosine, hypoxanthine and xanthine in response to ischemic-like condition. Selective O(2) or glucose deprivation, and Ca(2+)-free conditions differentially affected the outflow of [3H]norepinephrine and [3H]purines, indicating that they derived from different sources. The ABC transporter inhibitors glibenclamide (100 microM) and verapamil (100 microM) as well as low-temperature inhibited [3H]purine release evoked by ischemic-like conditions. Surgical denervation of the spleen reduced endogenous catecholamine content and [3H]norepinephrine uptake of the spleen, but not that of [3H]adenosine. In summary, these results demonstrate the release of norepinephrine and purines in response to an ischemic-like condition in an immune organ. Although both could provide an important source of extracellular catecholamines and purines involved at various levels of immunomodulation, the source and mechanism of norepinephrine and purine efflux seem different. PMID- 11063821 TI - Modulation of the IL-10/IL-12 cytokine circuit by interferon-beta inhibits the development of epitope spreading and disease progression in murine autoimmune encephalomyelitis. AB - IFN-beta has been shown to be effective in the treatment of multiple sclerosis (MS). However, the primary mechanism by which IFN-beta mediates its therapeutic effect remains unclear. Recent studies indicate that under defined conditions, IFN-beta may downregulate DC expression of IL-12. We and others have shown that IFN-beta may also downregulate IL-10. In light of the recently proposed paradigm that an IL-10/IL-12 immunoregulatory circuit controls susceptibility to autoimmune disease, we examined the effect of IFN-beta on the development and behavior of the autoreactive T cell repertoire during experimental autoimmune encephalomyelitis (EAE), an animal model sharing many features with MS. SWXJ mice were immunized with the immunodominant p139-151 determinant of myelin proteolipid protein (PLP), and at onset of EAE were treated every other day with IFN-beta. After eight weeks of treatment, we assessed autoreactivity and observed no significant IFN-beta effect on splenocyte proliferation or splenocyte production of IFN-gamma, IL-2, IL-4, or IL-5 in response to the priming determinant used to initiate disease. However, in IFN-beta treated mice, the cytokine profile in response to the priming immunogen was significantly skewed toward an increased production of IL-10 and a concurrent decreased production of IL-12. Moreover, the in vivo modulation of the IL-10/IL-12 immunoregulatory circuit in response to the priming immunogen was accompanied by an aborted development of epitope spreading. Our results indicate that IFN-beta induces a reciprocal modulation of the IL 10/IL-12 cytokine circuit in vivo. This skewed autoreactivity establishes an inflammatory microenvironment that effectively prevents endogenous self-priming thereby inhibiting the progression of disease associated with epitope spreading. PMID- 11063822 TI - Significant correlation between IL-10 levels and IgG indices in the cerebrospinal fluid of patients with multiple sclerosis. AB - We measured the interleukin (IL)-6 and IL-10 levels in the plasma and the cerebrospinal fluid (CSF) of a total of 23 relapsing multiple sclerosis (MS) patients [18 with conventional form of MS (C-MS) and 5 with optic-spinal form of MS (OS-MS)] using ELISA and correlated them with the IgG indices and oligoclonal IgG bands (OB) to determine whether these cytokines play a role in the intrathecal immunoglobulin production. IL-10 values in the CSF significantly correlated with the IgG indices and tended to be higher in OB-positive patients. In contrast, IL-10 values in the plasma and IL-6 values in the CSF and the plasma did not correlate with the IgG indices or OB. The CSF-IL-10 value in OS-MS were much lower than those of C-MS, but those of CSF IL-6 did not differ between C-MS and OS-MS. The results remained unchanged even when OS-MS patients were excluded. Our results may suggest a role of IL-10 in upregulating the intrathecal IgG synthesis in relapsing MS. PMID- 11063823 TI - Interleukin-1beta-dependent changes in the hippocampus following parenteral immunization with a whole cell pertussis vaccine. AB - Neurological side effects are a major cause of concern following immunization with a number of vaccines, especially the whole cell pertussis vaccine (Pw). In this study we report that IL-1beta concentrations were significantly increased in the hippocampus following subcutaneous (s.c.) injection of Pw, and that this was accompanied by increased activity of the stress-activated kinase, c-Jun-N terminal kinase (JNK) and a decrease in glutamate release. These effects were mimicked by s.c injection of active pertussis toxin (PT) or lipopolysaccharide (LPS). Incubation of hippocampal synaptosomes in the presence of Pw, PT or LPS also resulted in increased JNK activation and decreased glutamate release, effects which were mimicked by IL-1beta and blocked by the IL-1 receptor antagonist (IL-ra). Our observations are consistent with the hypothesis that IL 1beta induced by active bacterial toxins present in vaccine preparations, mediate the neurochemical and perhaps the neurological effects of Pw. PMID- 11063824 TI - Female sex steroids: effects upon microglial cell activation. AB - Multiple sclerosis occurs more commonly in females than males. However, the mechanisms resulting in gender differences in multiple sclerosis are unknown. Activated microglia are believed to contribute to multiple sclerosis pathology, perhaps in part due to production of nitric oxide (NO) and TNF-alpha, molecules which can be toxic to cells including oligodendrocytes. The current study demonstrates that the female sex steroids estriol, beta-estradiol and progesterone inhibit lipopolysaccharide (LPS) induction of nitric oxide (NO) production by primary rat microglia and by the mouse N9 microglial cell line. These hormones act by inhibiting the production of inducible nitric oxide synthase (iNOS) which catalyses the synthesis of NO. Estriol likely inhibits iNOS gene expression since the hormone blocks LPS induction of iNOS RNA levels. The pro-inflammatory cytokines IFN-gamma and TNF-alpha are believed to be important modulators of multiple sclerosis. Here, we demonstrate that estrogens and progesterone also inhibit NO production by microglial cells activated in response to these cytokines. Activated microglia elicit TNF-alpha in addition to NO and we further demonstrate that estrogens and progesterone repress TNF-alpha production by these cells. Finally, estriol and progesterone, at concentrations consistent with late pregnancy, inhibit NO and TNF-alpha production by activated microglia, suggesting that hormone inhibition of microglial cell activation may contribute to the decreased severity of multiple sclerosis symptoms commonly associated with pregnancy. PMID- 11063825 TI - Interferon-beta treatment in multiple sclerosis patients decreases the number of circulating T cells producing interferon-gamma and interleukin-4. AB - Systemic administration of interferon (IFN)-beta has been recently approved for the treatment of relapsing-remitting multiple sclerosis (RRMS). The immunological mechanism by which IFN-beta ameliorates MS is still partially unknown. We measured the number of blood circulating CD4(+), CD4(-), CD8(+), and CD8(-) T cells secreting IFN-gamma and IL-4 in 26 RRMS patients followed for up to 9 months of an alternate day s.c. treatment with 8x16 IU of IFN-beta1b. Compared to pre-treatment values, a significant (P<0.05) reduction of CD4(+), CD4(-), CD8(+) and CD8(-) cells producing IFN-gamma and of CD4(+) and CD4(-) cells producing IL 4 was observed in MS patients. The IFN-beta-associated effect was evident soon after the beginning of the treatment and persisted for the entire follow-up period. We did not observe any effect of IFN-beta treatment on the percentage of IL-4-producing CD8(+) and CD8(-) cells nor in that of natural killer (NK) cells producing IFN-gamma. Our results show that IFN-beta treatment in MS patients induces a profound and persistent down-regulation of the number of circulating T cells secreting IFN-gamma and IL-4 thus suggesting a broader rather than a specific immunomodulatory effect of IFN-beta in MS. PMID- 11063826 TI - Immunosuppression prevents neuronal atrophy in lupus-prone mice: evidence for brain damage induced by autoimmune disease? AB - An early onset of systemic, lupus-like disease in MRL-lpr mice is accompanied by deterioration in their behavioral performance and atrophy of pyramidal neurons in the parietal cortex and the hippocampal CA1 area. Using the immunosuppressive drug cyclophosphamide (CY) to attenuate the disease, we have tested the hypothesis that the autoimmune/inflammatory process is responsible for changes in brain morphology. A modified Golgi impregnation method revealed that, in comparison to saline-treated controls, immunosuppressive treatment with CY (100 mg/kg/week i.p. over 8 weeks) increased dendritic branching and spine numerical density in the CA1 region of MRL-lpr mice and MRL +/+ mice, which develop less severe manifestations of the disease. More interestingly, CY selectively prevented the atrophy and aberrant morphology of pyramidal neurons in the parietal cortex of MRL-lpr mice. The neuropathological measures (in particular reduced dendritic spine density) significantly correlated with increased serum levels of antinuclear antibodies and splenomegaly. The present results support the hypothesis that chronic autoimmune disease induces functionally important changes in neuronal morphology, and provide an empirical basis for understanding the behavioral dysfunction in systemic lupus erythematosus and autoimmune phenomena reported in some forms of mental illness. PMID- 11063827 TI - Expression of functional formyl peptide receptors by human astrocytoma cell lines. AB - Activation of astrocytes is important in the pathogenesis of a variety of diseases in the central nervous system, such as infection and neurodegeneration. We found that the bacterial chemotactic peptide, N-formyl-methionyl-leucyl phenylalanine (fMLF) induced potent migration and Ca(2+) mobilization in human astrocytoma cell lines. The effect of fMLF was pertussis toxin-sensitive, suggesting the involvement of seven transmembrane, G protein-coupled receptor(s) for fMLF. Scatchard analyses revealed that astrocytoma cell lines express both high- and low-affinity binding sites for [3H]fMLF. RT-PCR confirmed the expression of transcripts of fMLF receptors, the high-affinity FPR and the low affinity FPRL1 by these cells. Both fMLF and F peptide, a synthetic peptide domain of HIV-1 envelope protein which specifically activates FPRL1, increased secretion of IL-6 by astrocytoma cells. Our study demonstrates for the first time that FPR and FPRL1 expressed by astrocytoma cell lines are functional, and suggests a molecular basis for the involvement of these receptors in host defense in the brain. PMID- 11063828 TI - Astrocytes express functional chemokine receptors. AB - Multiple lines of evidence are presented characterizing the functional expression of chemokine receptors CXCR4, CCR1, CCR5, and CX3CR1 on astrocytes. Most of these receptors are expressed at low levels and may only be detectable on a subset of cells during disease or following cytokine induction. The expression of CXCR2, CCR2, CCR3, CCR10, CCR11, and several orphan receptors associated with HIV-1 infection has also been proposed. The appearance of several chemokine receptors implies a wider role for chemokines in the regulation of central nervous system functions. Available evidence indicates that selected chemokines induce further chemokine synthesis in astrocytes providing a mechanism to amplify inflammatory responses in the central nervous system. PMID- 11063829 TI - Persistent accumulation of cyclooxygenase-1 (COX-1) expressing microglia/macrophages and upregulation by endothelium following spinal cord injury. AB - Acute inflammation following spinal cord injury results in secondary injury and pathological reorganisation of the central nervous system (CNS) architecture. Cyclooxygenases (Prostaglandin Endoperoxide H Synthases, PGH) are key enzymes in the conversion of arachidonic acid into prostanoids which mediate immunomodulation, mitogenesis, apoptosis, blood flow, secondary injury (lipid peroxygenation) and inflammation. Here, we report cyclooxygenase-1 (COX-1) expression following spinal cord injury. In control spinal cords, COX-1 expression was localized by immunohistochemistry to ependymal cells, some neurons, inclusive dorsal and ventral root ganglion cells, few endothelial cells but rarely to brain microglia/macrophages. In injured spinal cords, COX-1(+) microglia/macrophages accumulated highly significantly (P<0.0001) at peri lesional areas and in the developing necrotic core early after injury. Here numbers of COX-1(+) cells remained persistently elevated up to 4 weeks following injury. Further, COX-1(+) cells were located in perivascular Virchow-Robin spaces, between spared axons and in areas of Wallerian degeneration. Double labeling experiments confirmed co-expression of COX-1 by ED-1(+) and OX-42(+) microglia/macrophages. Transiently after infarction most COX-1(+) microglia/macrophages coexpress the activation antigen OX-6 (MHC class II). However, the prolonged accumulation of COX-1(+) microglia/macrophages at the lesion site enduring the acute post injury inflammatory response points to a role of COX-1 in tissue remodeling or secondary injury. We have identified and localized persistent accumulation of COX-1 expressing cells which might be a potential pharmacological target following spinal cord injury. Therefore, we suggest that approaches based on: (i) short-term; and (ii) selective COX-2 blocking alone might not be a sufficient tool to suppress the local synthesis of prostanoids. PMID- 11063830 TI - Neurobehavioral alterations in mice with a targeted deletion of the tumor necrosis factor-alpha gene: implications for emotional behavior. AB - Tumor necrosis factor-alpha (TNF-alpha) is emerging as an important modulator of the function of the central nervous system. In the present study, we investigated a role of endogenous TNF-alpha in cognitive and emotional function using mice with targeted deletions of the TNF-alpha gene. TNF-alpha-(-/-) mice showed normal diurnal rhythms of spontaneous locomotor activity and cognitive functions. Emotional behavior in the mutant mice, however, was significantly altered, which manifested in the performance in the open-field, elevated plus maze, and forced swimming tests. The altered performance in the elevated plus maze test was significantly alleviated by treatment with diazepam. Postmortem brain analysis of TNF-alpha-(-/-) mice revealed a significant increase in serotonin metabolism in the brain. These findings suggest a role for endogenous TNF-alpha in emotional behavior, which may possibly be related to alterations of serotonine metabolism. PMID- 11063831 TI - Morphine suppresses complement receptor expression, phagocytosis, and respiratory burst in neutrophils by a nitric oxide and mu(3) opiate receptor-dependent mechanism. AB - We investigated whether morphine and fentanyl influence surface receptor expression, phagocytic activity and superoxide anion generation of neutrophils in a whole blood flow cytometric assay. Morphine suppressed complement and Fcgamma receptor expression and neutrophil function in a concentration- and time dependent manner. Morphine-induced changes were similar to those caused by the nitric oxide (NO) donor S-nitroso-N-acetyl-penicillamine and were abolished by preincubation with the NO synthase inhibitor N-nitro-L-arginine as well as naloxone. Fentanyl had no immunosuppressive effects. These results suggest that these neutrophil functions are inhibited by morphine-stimulated NO release mediated by the mu(3) opiate receptor subtype found on immunocytes. PMID- 11063832 TI - T-cell anti-apoptotic mechanisms in inflammatory myopathies. AB - Recent studies have shown an up-regulation of the Fas/Fas ligand system in inflammatory myopathies. In myositis, however, the major Fas-mediated cytotoxicity which activates caspases bypasses apoptosis. We therefore evaluated the expression of proteins promoting cell survival, such as bcl-2, bcl-x(l) and cyclin-dependent kinase inhibitors, on muscle biopsies from 14 patients with polymyositis, dermatomyositis, inclusion body myositis and HIV-associated myositis. Our data demonstrate that inflammatory cells are immunoreactive for bcl x(l), p16 and p57, three apoptosis-preventing proteins. Hence, we assume that these proteins might protect T cells from apoptotic nuclear changes. Our results could explain the non-self-limiting nature of inflammatory myopathies. PMID- 11063833 TI - Suppression of experimental autoimmune myasthenia gravis in IL-10 gene-disrupted mice is associated with reduced B cells and serum cytotoxicity on mouse cell line expressing AChR. AB - To analyze the role of interleukin-10 (IL-10) in experimental autoimmune myasthenia gravis (EAMG) pathogenesis, we induced clinical EAMG in C57BL/6 and IL 10 gene-knockout (KO) mice. IL-10 KO mice had a lower incidence and severity of EAMG, with less muscle acetylcholine receptor (AChR) loss. AChR-immunized IL-10 KO mice showed a significantly higher AChR-specific proliferative response, altered cytokine response, lower number of class II-positive cells and B-cells, but a greater CD5(+)CD19(+) population than C57BL/6 mice. The lower clinical incidence in IL-10 KO could be explained not by a reduction of the quantity, but by a possible difference in the pathogenicity of anti-AChR antibodies. PMID- 11063834 TI - Mechanism of nasal tolerance induced by a recombinant fragment of acetylcholine receptor for treatment of experimental myasthenia gravis. AB - Acetylcholine receptor (AChR) is the major autoantigen in myasthenia gravis (MG) and experimental autoimmune MG (EAMG). Here we analyze the mechanisms involved in suppression of ongoing EAMG in rats by nasal administration of a recombinant fragment from the human AChR alpha-subunit. We demonstrate that such a fragment, expressed without a fusion partner, confers nasal tolerance that can be adoptively transferred. Our observations suggest that the underlying mechanism of this nasal tolerance is active suppression involving a shift from a Th1 to a Th2/Th3-regulated AChR-specific response which may be mediated by down regulation of costimulatory factors. PMID- 11063835 TI - Complement activation by titin and ryanodine receptor autoantibodies in myasthenia gravis. A study of IgG subclasses and clinical correlations. AB - To elucidate the mechanism of immune damage caused by titin and ryanodine receptor (RyR) autoantibodies in myasthenia gravis (MG), we studied the complement-activating capacity and the IgG subclass distribution of these autoantibodies in sera from 49 MG patients. Complement activation occurred in 38 out of 49 titin antibody positive sera, and in 14 out of 21 RyR antibody positive sera. The titin antibodies occurred only in the IgG 1 and IgG 4 subclasses, whereas the RyR antibodies occurred in all four IgG subclasses but with IgG 1 predominance. Complement-activating RyR antibodies occurred with higher frequency in sera of thymoma MG than of late-onset MG. RyR IgG 1 antibodies occurred more often in severe MG than in mild and moderate disease groups. Mean total IgG and IgG 1 titin and RyR antibody titers fell during long-time patient observation together with an improvement of the MG symptoms. PMID- 11063836 TI - Antigen-presenting capability of glial cells under glioma-harboring conditions and the effect of glioma-derived factors on antigen presentation. AB - The antigen-presenting capability of syngeneic rat glial cells was investigated under glioma-harboring conditions. Microglia induced a significant proliferation of glioma-primed splenocytes, but astrocytes did not. Furthermore, astrocytes suppressed the accessory cell function of microglia. The presence of both indomethacin and anti-interleukin (IL)-10 neutralizing antibody during priming of microglia enhanced splenocyte proliferation. The glioma culture supernatants down regulated the interferon-gamma-induced expression of major histocompatibility complex class II molecules on microglia. The down-regulation was blocked by indomethacin and anti-IL-10 antibody. The results suggest that microglia but not astrocytes may function as antigen-presenting cells in glioma, and that glioma may suppress the antigen-presenting abilities of microglia. PMID- 11063837 TI - VLA-4/CD49d downregulated on primed T lymphocytes during interferon-beta therapy in multiple sclerosis. AB - Effects on adhesion molecules of immune cells might contribute to the mode of action of interferon-beta (IFN-beta) in multiple sclerosis (MS). We have serially monitored the cell surface expression of integrins CD49d (VLA-4) and CD11a (LFA 1) on fresh T lymphocyte subpopulations from 5 MS patients monthly for 2 months prior to treatment and for 3 months on treatment with IFN-beta1b. In parallel, we assessed inflammatory disease activity by monthly contrast-enhanced magnetic resonance imaging (MRI). IFN-beta treatment specifically downregulated CD49d expression on CD8+ and CD4+/CD45RO+ 'memory' T lymphocytes and differentially modulated the proportion of CD4+, CD8+ and CD27+ T cells. These effects may play an important role in the reduction of central nervous system cell trafficking and inflammation in MS. PMID- 11063838 TI - Anti-striatal antibodies in Tourette syndrome cause neuronal dysfunction. AB - Serologic studies of children with Tourette syndrome (TS) have detected anti neuronal antibodies but their role in TS has not been explored. Stereotypies and episodic utterances, analogous to involuntary movements seen in TS, were induced in rats by intrastriatal microinfusion of TS sera or gamma immunoglobulins (IgG) under noninflammatory conditions, as found in TS. Immunohistochemical analysis confirmed the presence of IgG selectively bound to striatal neurons. These data support the hypothesis that binding of an anti-neuronal antibody from some children with TS induced striatal dysfunction and suggest a possible cause for the basal ganglia alterations observed in children with TS. PMID- 11063839 TI - Increased cellular expression of the caspase inhibitor FLIP in intrathecal lymphocytes from patients with multiple sclerosis. AB - Failure of Fas-mediated apoptosis of potentially pathogenic, autoreactive T lymphocytes may be involved in the pathogenesis of multiple sclerosis. The intracellular protein FLIP, a naturally occurring caspase-antagonist, is a potent inhibitor of the Fas signalling pathway that may block Fas-mediated apoptosis of activated lymphocytes. This study reports specific overexpression of both long and short forms of FLIP in intrathecal lymphocytes from patients with multiple sclerosis. The overexpression of FLIP is independent of cellular expressions of Fas receptor or the anti-apoptotic protein Bcl-2. These results provide a better understanding of some of the intrinsic immunoregulatory mechanisms that are involved in multiple sclerosis. PMID- 11063840 TI - MHC antigen expression in human first trimester spinal cord with implications for clinical transplantation procedures. AB - We report human leukocyte antigen (HLA) class I expression in 5-17% and class II in 0-9% of first trimester human spinal cord cells. After 8 days in culture with gamma-interferon, >87% of the spinal cord cells expressed HLA class II. However, mixed cultures of adult human peripheral lymphocytes and immature human spinal cord cells, showed no induction of lymphocyte proliferation prior to or after gamma-interferon exposure in culture. In conclusion, we report non-immunogenic expression of HLA antigens in the human first trimester spinal cord. PMID- 11063841 TI - CD4(+)CD45RO(+)CD49d(high) cells are involved in the pathogenesis of relapsing remitting multiple sclerosis. AB - In the animal model of multiple sclerosis, experimental autoimmune encephalomyelitis, encephalitogenic T cells differ from the non-encephalitogenic ones in their expression of CD49d. The disease-inducing CD49d(high) and not the CD49d(low) cells enter the brain parenchyma. In this context, we characterized CD4(+)(CD45RO(+))CD49d(high) cells in relapsing-remitting multiple sclerosis (RR MS) patients. These cells, showing characteristics of activated cells able to produce pro-inflammatory cytokines, were found to be increased in peripheral blood during relapses and present in high numbers in cerebrospinal fluid. These results suggest that the CD4(+)CD45RO(+)CD49d(high) subpopulation in RR-MS patients includes autoreactive cells and may be target for immunotherapy. PMID- 11063842 TI - Increased peripheral blood interferon gamma-producing T cells in acute disseminated encephalomyelitis. AB - There have been few reports on immunological studies in patients with acute disseminated encephalomyelitis (ADEM). We investigated the immunological features of ADEM using flow cytometry to examine interferon gamma (IFN-gamma)-and interleukin 4 (IL-4)-producing peripheral blood CD3+T cells from four patients with ADEM, three other neurological disorders (Fisher syndrome, epilepsy and aseptic meningitis) and 10 healthy children. IFN-gamma-producing CD3+T cells were increased in ADEM during the acute stage. In a relapsing case of ADEM, the percentages of IFN-gamma-producing CD3+T cells correlated with disease activity. There were no significant changes of IL-4-producing CD3+T cells in ADEM during the acute and convalescent stages. In conclusion, peripheral blood IFN-gamma producing T cells are related to the pathogenesis at the early phase of the acute ADEM. PMID- 11063843 TI - A case of Guillain-Barre syndrome following a family outbreak of Campylobacter jejuni enteritis. AB - We describe an outbreak of Campylobacter jejuni enteritis involving three family members of whom one developed Guillain-Barre syndrome (GBS). The patients' serum reacted strongly with several gangliosides and with the lipopolysaccharide (LPS) fractions from the C. jejuni strains isolated from his family members. Only low titer anti-ganglioside antibodies were found in his siblings. HLA-typing did not indicate a locus associated with auto-antibody production. Comparing the immune response in GBS patients and C. jejuni enteritis patients can be of great value in determining the additional factors that lead to post-Campylobacter GBS. Ganglioside mimicry alone is necessary but not sufficient for the induction of anti-ganglioside antibodies. Other susceptibility factors are required to induce an anti-neural immune response. PMID- 11063844 TI - Lesion associated expression of urokinase-type plasminogen activator receptor (uPAR, CD87) in human cerebral malaria. AB - Blood-brain barrier disintegration and inflammatory cell recruitment are key processes in the pathogenesis of cerebral malaria (CM). Recent data provide convincing evidence that the serine protease urokinase-type plasminogen activator receptor (uPAR) is a key molecule in promoting cell adhesion and spreading. We have now analyzed expression of urokinase-type plasminogen activator receptor (uPAR, CD87), which is part of a cell surface associated proteolytic system, in brains of eight CM patients and seven neuropathologically unaltered and diseased controls by immunohistochemistry. Double labeling experiments with antibodies directed against CD68 (macrophages/microglial cells), myeloid-related protein (MRP8), and glial fibrillary acid protein (GFAP) confirmed the nature of uPAR expressing cells. We observed focal accumulation of uPAR expressing macrophages/microglial cells in Durck's granulomas and adjacent to petechial hemorrhages, in astrocytes, and in endothelial cells. In contrast, focal uPAR expression in macrophages/microglial cells but not in astrocytes was found in microglial nodules of toxoplasmic encephalitis and in the cellular infiltrate of bacterial meningitis. Normal brains showed only faint uPAR expression in endothelial cells. We conclude from these data that lesion-associated uPAR expression at least in part contributes to blood-brain barrier alteration and immunologic dysfunction in CM patients. PMID- 11063845 TI - Demonstration of anti-HuD specific oligoclonal bands in the cerebrospinal fluid from patients with paraneoplastic neurological syndromes. Qualitative evidence of anti-HuD specific IgG-synthesis in the central nervous system. AB - The presence of HuD-specific oligoclonal IgG bands in the CSF was investigated in five patients with paraneoplastic neurological syndromes. All patients revealed intrathecal synthesis of HuD specific antibodies in the CSF, as estimated from elevated antibody indices (>1.5) in an IgG-ELISA using recombinant HuD-protein as antigen. Isoelectrofocussing combined with affinity blotting showed reactivity of IgG bands with recombinant HuD antigen in all CSF samples. These data support the idea that HuD specific antibodies in the CSF are produced mainly by B-cell clones in the central nervous system. These findings support the hypothesis of autoimmunity in the pathogenesis of anti-Hu associated paraneoplastic neurological syndromes. PMID- 11063847 TI - A new name for parasitology today AB - Parasitology Today is officially the Number One ranked journal in parasitology (impact factor 4.940). Since its launch in 1985, Parasitology Today has been part of the Trends Journal series - the most highly cited group of monthly review journals in the world, that includes Trends in Biochemical Sciences, Trends in Microbiology, Trends in Genetics, Immunology Today and Trends in Cell Biology. PMID- 11063846 TI - Myelin oligodendrocyte glycoprotein (MOG) polymorphisms and adrenoleukodystrophy. AB - We describe four novel sequence variants in the Myelin Oligodendrocyte Glycoprotein (MOG) gene. A total of six sequence variants of the MOG gene were identified in eleven out of 44 ALD patients investigated: G15A, CTC repeat in exon 1, Val142Leu, Val145Ile, 551+68A-->G and 551+77C-->T. Screening studies demonstrated that all these polymorphisms are present in 50 unaffected control male individuals of the same population and in the different phenotypes of ALD patients, indicating that they do not contribute to phenotype variability in ALD. PMID- 11063848 TI - Oswaldo Cruz meets Charles Darwin. PMID- 11063849 TI - A career of many colors. PMID- 11063850 TI - What is the real target for ivermectin resistance selection in Onchocerca volvulus? PMID- 11063852 TI - Errata PMID- 11063851 TI - Species-transcending regulation of malaria parasitaemia. PMID- 11063854 TI - Patch clamping the malaria-infected red blood cell PMID- 11063853 TI - All that cycles may not Be climate-driven PMID- 11063855 TI - Chickens, malaria and zoonoses on the Web PMID- 11063856 TI - Parasites and behaviour: an ethopharmacological perspective. AB - Ethopharmacology combines an ethological approach to the understanding of the causes and functions of behaviour with pharmacological analysis of the underlying neuromodulatory mechanisms. Recently, this approach has been applied to the analysis of the responses of animals to parasitized individuals and to the effects of parasites on various host behaviours (eg. host defences, mate responses) and their neurobiological correlates. Martin Kavaliers, Douglas Colwell and Elena Choleris explain here how ethopharmacology can be used to address the mechanisms that underlie the often subtle effects of parasites on host behaviour. PMID- 11063857 TI - Malaria-related anaemia. AB - Malaria infection in humans by Plasmodium species is associated with a reduction in haemoglobin levels, frequently leading to anaemia. Plasmodium falciparum causes the most severe and profound anaemia, with a significant risk of death. This cannot be explained simply by the direct destruction of parasitized red blood cells at the time of release of merozoites, a process shared by all these species. In this review, Clara Menendez, Alan Fleming and Pedro Alonso focus on recent advances in our knowledge of the pathophysiology, epidemiology, management and prevention of anaemia from falciparum malaria. PMID- 11063858 TI - The role of mucins in host-parasite interactions. Part I-protozoan parasites. AB - Parasite-derived mucin-like molecules might be involved in parasite attachment to and invasion of host cells. In addition, parasites might secrete mucin-degrading enzymes, enabling the penetration of protective mucus gels that overlie the mucosal surfaces of their potential hosts. Furthermore, they might generate binding ligands on the membrane-bound mucins of host cells by using specific glycosidases. It is possible that host mucins and mucin-like molecules prevent the establishment of parasites or facilitate parasite expulsion. They might also serve as a source of metabolic energy and adhesion ligands for those parasites adapted to exploit them. Sally Hicks and colleagues here review the biochemical properties of mucins and mucin-like molecules in relation to interactions (established and putative) between protozoan parasites and their hosts. PMID- 11063859 TI - Metabolic aspects of glycosomes in trypanosomatidae - new data and views. AB - The energy metabolism of Trypanosomatidae has been the subject of many reviews during the past decade. In recent years, however, new data have led to a more complete picture of trypanosomatid metabolism and a reappraisal of the role of some characteristic organelles in the energy supply of these parasites. For years, the glycosome was thought to be a peroxisome-like organelle that had evolved to allow the parasites to carry out glycolysis at a high rate using a relatively small amount of enzyme. However, the results of recent studies of trypanosomatid glycolysis and the detection of various other pathways and enzymes in the organelle necessitate a modification of this view. Here, Paul Michels, Veronique Hannaert and Frederic Bringaud review the new data and discuss the possible implications for our view on the role of the glycosome. PMID- 11063860 TI - Proteophosphoglycans of Leishmania. AB - Proteophosphoglycans are an expanding family of highly glycosylated Leishmania proteins with many unusual and some unique structural features. The novel protein glycan linkage in proteophosphoglycans - phosphoglycosylation of Ser by lipophosphoglycan-like structures - emerges as a major form of protein glycosylation in Leishmania. Here, Thomas Ilg reviews the chemical structure, the ultrastructure, the genes and the potential functions of different members of this novel family of parasite glycoproteins. PMID- 11063861 TI - Defining a schistosomiasis vaccination strategy - is it really Th1 versus Th2? AB - Successful vaccine development for schistosomiasis has been hindered by a lack of consensus on the type of immune response that would provide maximum levels of protective immunity and incomplete knowledge of the key antiparasite effector mechanisms. Many vaccine studies conducted in mice support type-1-cytokine mediated effector mechanisms, while acquired resistance in humans correlates with type-2-cytokine-mediated responses. However, recent data from cytokine-knockout mice suggest that choosing between these opposing pathways may be less important than previously hypothesized, as discussed here by Thomas Wynn and Karl Hoffmann. PMID- 11063862 TI - Transport of critically ill patients: we can do better! PMID- 11063863 TI - Condition on arrival of transferred critically ill patients. AB - We performed a retrospective inventory of the condition of transferred patients to our 11-bed medical ICU, aimed firstly to measure the quality of these transports and secondly to identify variables that may predict a high risk of deterioration during transferral. By a search in our hospital database, we identified 112 consecutive patients (47 women/65 men) transferred from other hospitals (distance 20-350 km) to our ICU over a period of 14 months. The following data were collected on departure (if available) and on arrival: blood pressure, heart rate, temperature, oxygen saturation, routine laboratory parameters, arterial blood gas analysis, lactic acid, settings of mechanical ventilation, use of vasopressor/inotropic medication, presence of venous and arterial catheters and Apache II score on arrival. No major worsening during transportation was found, looking at the whole group. However, individual data showed severe deterioration of some patients during transport. We were not able to point out parameters that could predict hemodynamic or respiratory instability during transport or condition on arrival. In conclusion, quality of transport seems fairly good; in individual cases, improvements are possible. Therefore, we plan to investigate whether or not a strict protocol, based on recommendations in the literature and on local feasibility can further improve condition on arrival and survival of transferred ICU patients in our adherence region. PMID- 11063864 TI - Acute pyelonephritis: a cause of acute renal failure? AB - Two patients with acute renal failure due to acute pyelonephritis are described. Examination of the renal biopsy showed normal glomeruli, severe interstitial neutrophilic infiltration and edema with no signs of acute tubular necrosis. Until now, only twelve biopsy-proven proven cases have been reported. A review of the literature on acute renal failure due to acute pyelonephritis is presented. PMID- 11063865 TI - Hepatic steatosis and lactic acidosis caused by stavudine in an HIV-infected patient. AB - Lactic acidosis and hepatic steatosis caused by mitochondrial toxicity of nucleoside reverse transcriptase inhibitors (NRTI) is a rare cause of liver disease with a high mortality rate. This report describes a male, HIV-positive patient with a 4-week history of nausea, vomiting and abdominal pain. His medication consisted of prednisone 5 mg od (because of auto-immune thrombocytopenia), didanosine (for 2 years) and stavudine (for 3 months). Laboratory studies showed cholestasis and elevation of aminotransferases. Lactic level was not measured. Liver biopsy revealed steatosis and cholestatic hepatitis. In the absence of other causes of liver disease a probable diagnosis of stavudine-induced hepatic toxicity was made. After discontinuation of NRTI, he recovered completely. Because lactic acidosis had not been confirmed, stavudine was restarted and within 1 week the lactate level increased significantly. Therefore stavudine was discontinued again. One year later the patient is doing well on a double protease inhibitor regimen. In conclusion, clinicians treating patients with NRTI should be aware of the risk of lactic acidosis and hepatic steatosis. When this is suspected, all NRTI must be stopped. The diagnosis can be made when elevated lactate levels and hepatic steatosis are present in the absence of other causes of liver disease. PMID- 11063866 TI - Smoking cessation: which aids are worthwhile? AB - Smoking is a major preventable health risk in western society. In the Netherlands, it is held responsible for 86 and 36% of annual mortality from lung cancer and cardiovascular disease, respectively. Nevertheless about 33% of Dutch people smoke. Only 2% of smokers quit successfully after being advised to stop once by a physician. Although the medical profession should play a leading role in campaigns to stop smoking, general practitioners advise only 10% of their smokers to quit. An overview was made of the various aids that can be used to support attempts to quit smoking. Three aids: supportive schedules, nicotine replacement and bupropion chloride had proven long-term effectiveness in up to 5 10, 3-13 and 11-15% of the subjects, respectively. In conclusion, supportive counselling combined with nicotine substitutes or bupropion chloride is the most worthwhile intervention to support quitting attempts. Wider application of this strategy is expected to have major implications on morbidity (50% myocardial infarct risk reduction) and mortality in the Netherlands. PMID- 11063867 TI - Current status of umbilical cord blood hematopoietic stem cell transplantation. AB - The number of umbilical cord blood transplants is increasing worldwide. At this time, it is important to evaluate their results and to compare the outcome of umbilical cord blood transplants with allogeneic bone marrow transplants. Data have been reported to the Eurocord Registry by multiple transplant centers. Close links have been established with the cord blood banks through Netcord. Bone marrow transplant data have been provided by transplant centers and through the European Blood and Marrow Transplant (EBMT) and International Bone Marrow Transplant Registries (IBMTR). Eurocord has analyzed the outcomes of 527 umbilical cord blood transplants from 121 transplant centers and 29 countries. The donor was related in 138 cases and unrelated in 399 cases. The results showed that survival with umbilical cord blood transplants was comparable to that with related or unrelated bone marrow transplants. Engraftment with cord blood was delayed resulting in an increased incidence of early transplant complications. The incidence of acute and chronic graft-vs-host disease was reduced with cord blood grafts even in HLA-mismatched transplants and in adults. In patients with leukemia, the rate of relapse was similar to the rate of relapse after bone marrow transplant. The overall event-free survival with umbilical cord blood transplantation was not statistically different compared to bone marrow transplants. This large registry study confirms the potential benefit of using umbilical cord blood hematopoietic stem cells for allogeneic transplants. PMID- 11063868 TI - mNotch1 signaling reduces proliferation of myeloid progenitor cells by altering cell-cycle kinetics. AB - Notch receptors are involved in the regulation of cell-fate decisions, differentiation, and proliferation in many tissues. The expression of Notch receptors on hemopoietic cells and of cognate ligands on bone marrow stromal cells suggests a possible role for Notch signaling in the regulation of hemopoiesis. We were interested to assess the involvement of Notch1 signaling on cell proliferation of myeloid progenitor cells. Proliferation, cell-cycle status, and apoptosis of myeloid progenitor 32D cell lines engineered to permit the conditional induction of the constitutively active intracellular domain of mNotch1 (mN1(IC)) by the 4-hydroxytamoxifen(OHT)-inducible system were analyzed in the presence or absence of OHT. The induction of mN1(IC) by OHT resulted in reduction of proliferation (p<0.01) and accumulation of cells in the G(1)/G(0) phase of the cell cycle (p<0.001) without substantially affecting apoptosis of 32D cells. These effects were observed under culture conditions that allow differentiation and, to a lesser degree, under conditions that normally promote self-renewal in the absence of differentiated cells. Our data suggest that mNotch1 signaling suppresses proliferation of myeloid progenitor cells by altering cell-cycle kinetics. PMID- 11063869 TI - Gene expression in chronic lymphocytic leukemia B cells and changes during induction of apoptosis. AB - Our studies in chronic lymphocytic leukemia (CLL) are directed at understanding which signals maintain viability in vivo and become lost upon removal of leukemic cells from the body, such that they immediately begin to undergo apoptosis ex vivo. In this report, we examine changes in gene expression observed between freshly isolated CLL B cells and after maintenance in vitro with and without Fludara. We compare these effects with an Epstein-Barr virus (EBV)-transformed cell line treated similarly. Kinetic effects of drug treatment on apoptosis and cell division were examined with DNA laddering, radioisotopic labeling, and flow cytometry using the fluorescent dye carboxyfluorescein diacetate succinimidyl ester. Reverse transcriptase polymerase chain reaction and hybridization blots of microarray cDNA analyses were performed to examine gene expression. We demonstrate that many genes, especially cyclin D1, were downregulated after culture of CLL cells. Anti-apoptotic genes BAG-1 and Akt2 were upregulated. The greatest positive effect with Fludara was the upregulation of JNK1. The EBV transformed cell line was resistant to classic DNA laddering induced with Fludara. Although DNA synthesis was blocked, the EBV-transformed cell line had some ability to recover from treatment following drug washout. CLL cells express cell cycle regulatory genes that are specific for activated cells in the G(1)-S phase of the cell cycle. Growth regulatory signals are lost when the leukemic cells are isolated from the body. Fludara enhances kinetics of apoptosis and induces expression of a gene responsive to stress that regulates expression of a kinase involved in initiation of the apoptotic pathway. PMID- 11063870 TI - Adoptive immunotherapy with haploidentical allogeneic peripheral blood lymphocytes following autologous bone marrow transplantation. AB - Patients who undergo autologous bone marrow transplantation for acute leukemia are at high risk for relapse. We have evaluated the feasibility of administering cell-mediated immunotherapy with family-related haploidentical lymphocytes following autologous bone marrow transplantation in order to evoke a graft-vs leukemia effect in the autologous setting.Twenty-six patients aged 1.5-48 years were enrolled in this study. Eighteen suffered from acute myeloid leukemia, seven from acute lymphoblastic leukemia, and one from myelodysplastic syndrome. Eleven patients were transplanted in first remission, six in second remission, one in fourth remission, and eight in relapse. Conditioning consisted of Busulfan/Cyclophosphamide or Busulfan/Thiotepa/Cyclophosphamide. Nineteen patients (Group A) were treated with gradual increments of haploidentical donor T cells, starting on day +1, with an additional course of T cells plus intravenous recombinant human interleukin-2 one month later if no signs of graft-vs-host disease developed in the interim. Seven patients (Group B) were treated with high dose haploidentical T cells on day +1 in conjunction with intravenous recombinant human interleukin-2. Donor cells were detected in the peripheral blood of both groups 12-48 hours post-cell-mediated immunotherapy, peaking at 48 hours. Three patients in Group A developed transient Grade I graft-vs-host disease. One patient in Group B developed Grade I, and three Grade IV, graft-vs-host disease. Group A patients engrafted normally, but the Group B patients with Grade IV graft vs-host disease showed no signs of engraftment. Our results show that it is feasible to induce graft-vs-host disease in the autologous stem cell transplantation setting. However, the high-dose regimen of haploidentical T cells in conjunction with interleukin-2 results in severe toxicity and nonengraftment. PMID- 11063871 TI - Ikaros expression in human hematopoietic lineages. AB - The Ikaros gene has been implicated in lymphoid development and proliferation from the results of gene targeting studies in mice. Recently we reported that the Ikaros gene may be involved in the disease progression of chronic myelogenous leukemia (CML). In this report, we investigated Ikaros isoforms in human non lymphoid leukemia cell lines and normal granulocyte/macrophage (CFU-GM) and erythroid (BFU-E)-derived colonies. We evaluated Ikaros gene expression by RT PCR, Southern blotting, sequencing analysis, Northern blotting, and immunoblotting.Ikaros isoforms Ik-1 and Ik-2, 3 were predominantly expressed in human non-lymphoid leukemia cell lines. Ik-4 and Ik-8 were also detectable as a minor population. In contrast to the previous report in mice, multiple Ikaros isoforms were expressed in human CFU-GM and BFU-E-derived colonies, and the dominant-negative isoform Ik-6 was not detectable. We also showed that human Ikaros isoforms contained an additional coding sequence in the N-terminal region, which was highly homologous to the sequence reported in mice. These observations suggest that the Ikaros gene may play some role in the development of human non lymphoid lineage hematopoiesis. Moreover, the finding that the dominant-negative isoform Ik-6, which was overexpressed in patients with blast crisis of CML, was rarely detectable in non-lymphoid lineages supports its pathogenetic role in human hematologic malignancies. PMID- 11063872 TI - An inhibitor of PI3-K differentially affects proliferation and IL-6 protein secretion in normal and leukemic myeloid cells depending on the stage of differentiation. AB - In this study, we examined the involvement of the phosphatidylinositol 3-kinase (PI3-K) and p70S6 kinase signal transduction pathway in the interleukin-1(IL-1) mediated proliferation and cytokine production by normal and leukemic myeloid cells. Total AML blast populations, early progenitor (CD34(+)/CD36(-)) cells, and more differentiated (CD34(-)/CD36(+)) cells were treated with the PI3-K inhibitor Ly294002 and p70S6K inhibitor rapamycin. The effects on proliferation, IL-6 protein secretion, and intracellular signaling cascades were determined and compared with normal CD34(+) cells and monocytes. The function of the PI3-K pathway was dependent on the differentiation state of the AML cell population. In immature blasts, the IL-1-induced proliferation was strongly inhibited by Ly294002 and rapamycin, without a distinct effect on IL-6 protein production. In contrast, in mature monocytic blast cells inhibition of the PI3-K signaling route had a stimulatory effect on IL-6 protein secretion. Interestingly, these findings were not specifically linked to the malignant counterpart but were also observed with normal CD34(+) sorted cells vs mature monocytes. Evidence is provided that the Ly294002-induced increase in IL-6 protein secretion is linked to the cAMP dependent signaling pathway and not to changes in the phosphorylation of ERK or p38. However, although the enhanced IL-6 protein secretion is cAMP dependent, it was not found to be mediated by protein kinase A (PKA) or by the GTP-ase Rap1. This study indicates that inhibition of the PI3-K signaling pathway has an inhibitory effect on cell proliferation but a stimulatory effect on IL-6 expression mediated by a cAMP-dependent but PKA-independent route. PMID- 11063873 TI - Adaptor protein SKAP55R is associated with myeloid differentiation and growth arrest. AB - Activation of the SRC family of protein tyrosine kinases is an important component of intracellular signaling in hematopoiesis, but their critical substrates are less well understood. In this report, we describe the cloning and functional characterization of murine SKAP55R (mSKAP55R), an SRC family kinase substrate. Expression of mSKAP55R was examined by Northern blot. Phosphorylation of mSKAP55R was examined by transient transfection of COS cells. For overexpression studies, mSKAP55R was cloned into a bicistronic murine stem cell virus-based retrovirus. Transduced cells (FDC-P1 cell line and murine bone marrow) were FACS isolated by expression of the selectable marker green fluorescent protein.mSKAP55R showed 90% amino acid identity to the recently published human SKAP55R. mSKAP55R contained a central pleckstrin homology domain, a C-terminal SH3 domain, and a putative SRC kinase consensus substrate DEIY(260). mSKAP55R was expressed in all hematopoietic lineages, with relative mRNA levels greatest in cells of the myeloid and erythroid lineages. Induced myeloid differentiation of M1 and HL-60 cell lines was associated with an eight-fold increase in mSKAP55R mRNA. Transient expression of mSKAP55R in COS cells demonstrated that tyrosine 260 was the predominant site of phosphorylation by FYN kinase. Furthermore, this phosphotyrosine was essential for coimmunoprecipitation of FYN with mSKAP55R. Enforced expression of mSKAP55R inhibited in vitro growth of the myeloid FDC-P1 cell line and primary hematopoietic progenitors. In contrast, a tyrosine 260 mutant mSKAP55R had no effect on in vitro growth. These studies implicate mSKAP55R in the processes of myeloid differentiation and growth arrest. PMID- 11063874 TI - Globotriaosyl ceramide (CD77/Gb3) in the glycolipid-enriched membrane domain participates in B-cell receptor-mediated apoptosis by regulating lyn kinase activity in human B cells. AB - The role of CD77 expressed on a fraction of germinal center B cells, also known as glycosphyngolipid Gb3, and as a functional receptor for Shiga toxins (Stx) in B-cell receptor (BCR)-mediated apoptosis was investigated. Using Stx1-sensitive Burkitt's lymphoma Ramos cells as an in vitro model of CD77(+) germinal center B cells, intracellular signaling events mediated by either Stx1 or anti-CD77 antibody were examined immunobiochemically and immunocytologically. We observed prompt activation of Lyn and Syk kinases leading to increased binding of these proteins to surface IgM (sIgM) in Ramos cells after Stx1 treatment. We also observed microscopic colocalization of CD77 and sIgM after stimulation with Stx1. Along with the synergism between the cross-linking of CD77 and that of sIgM in their effect on apoptosis induction, it was highly probable that CD77 cross linking induces activation of the BCR signaling cascade. Analysis using sucrose density gradient centrifugation suggested that Stx1 binding to CD77 induced recruitment and activation of Lyn in the glycolipid-enriched membrane (GEM) fractions. Once activated, however, Lyn seemed to acquire an increased detergent solubility and moved outside of the GEM fractions. This study describes the participation of the GEM domain in BCR-signaling cascade and suggests a possible role of CD77 as a regulator of BCR-induced apoptosis in human B cells. PMID- 11063875 TI - Developmental changes of CD34 expression by murine hematopoietic stem cells. AB - It has been reported that fetal murine hematopoietic stem cells are CD34(+), whereas adult stem cells are CD34(-). We sought to delineate the developmental changes of CD34 expression by hematopoietic stem cells and carried out systematic analysis of long-term engrafting cells in the bone marrow and/or blood of perinatal, juvenile, and adult mice. To obtain information on the total population of stem cells, we prepared CD34(+) and CD34(-) populations of mononuclear cells without prior enrichment and assayed their long-term reconstituting abilities by transplantation into lethally irradiated Ly-5 congenic mice. All stem cells from perinatal to 5-week-old mice were CD34(+). In 7-week-old mice, CD34(-) stem cells began to emerge, and the majority of the stem cells were CD34(-) in the 10- and 20-week-old mice. Approximately 20% of adult stem cells expressed CD34. Developmental changes of CD34 expression from the positive to the negative state takes place between 7 and 10 weeks of age for the majority of murine stem cells. Approximately 20% of adult stem cells remain CD34(+). These observations provide insight into the current controversy regarding CD34 expression by adult hematopoietic stem cells and suggest that the majority of stem cells in human umbilical cord blood and bone marrow of young children are CD34(+). PMID- 11063876 TI - Differential MMP and TIMP production by human marrow and peripheral blood CD34(+) cells in response to chemokines. AB - As stromal cell-derived factor-1 (SDF-1), macrophage inflammatory protein-1alpha (MIP-1alpha), and interleukin-8 (IL-8) are implicated in the homing and mobilization of human hematopoietic progenitors (HPC), we hypothesized that these chemokines mediate the migration of HPC across subendothelial basement membranes by regulating production of matrix metalloproteinases (MMPs) and their natural tissue inhibitors (TIMPs). Assays for migration across reconstituted basement membrane (Matrigel) and chemotaxis were carried out using CD34(+) cells derived from normal human bone marrow (BM) and mobilized peripheral blood (PB). Secretion of MMPs and TIMPs was evaluated by zymography and reverse zymography and gene expression by RT-PCR. We found that an SDF-1 gradient increases the chemotaxis of BM and PB CD34(+) cells across Matrigel (BM > PB), which is blocked by inhibitors of MMPs (o-phenanthroline, rhTIMP-1, rhTIMP-2, and anti-MMP-9 and anti-MMP-2 antibodies) but enhanced by tumor necrosis factor-alpha (TNF-alpha), a strong stimulator of MMPs. Preincubation of these cells with SDF-1 stimulated the secretion of MMP-2 and MMP-9 in BM and PB CD34(+) cells but of TIMP-1 and TIMP-2 only in PB CD34(+) cells. Preincubation with MIP-1alpha and IL-8 also stimulated the secretion of MMP-9 and MMP-2 (BM > PB), but with respect to TIMPs, the effect was reversed (PB > BM), resulting in trans-Matrigel migration of BM but not of PB CD34(+) cells. We therefore propose that MMPs and TIMPs are involved in 1) SDF-1 induced chemotaxis of human HPC across subendothelial basement membranes, and 2) MIP-1alpha- and IL-8-stimulated migration of HPC. PMID- 11063877 TI - Identification of four human cDNAs that are differentially expressed by early hematopoietic progenitors. AB - The molecular processes that maintain the stem cell pool are largely unknown. Using polymerase chain reaction-driven subtraction, we examined genes that are differentially expressed by early hematopoietic progenitors. We expected that identifying genes that are uniquely expressed by the earliest precursors would provide insight into the mechanism(s) through which stem cell number is maintained and differentiation is regulated. Using CD34(+)CD38(-) cells as starting material, we identified four mRNAs, expressed by these cells, that are either absent or present in reduced amounts in more mature CD34(+)CD38(+) cells. One of these cDNAs (C40) encodes a known member of the subfamily of protein phosphatases (CL100) that exhibits dual substrate specificity for phosphotyrosine and phosphoserine/threonine-containing substrates and specifically inactivates MAP kinases. This phosphatase has been shown to play a role in regulating the differentiation of several cell types. The second cDNA (C23) is identical to LR11 (gp250), a member of the low-density lipoprotein receptor family. LR11 is unusual in that, in addition to 11 ligand-binding repeats, it contains a series of fibronectin type III repeats near its carboxyl terminal end that are similar to those found in cytokine receptors. It is highly expressed in developing brain, but hematopoietic expression has not been reported. The 178-bp fragment that we originally cloned is part of a 4,145-bp 3' untranslated region (UTR) that had not been previously sequenced and is among the largest human 3' UTRs ever reported. The other isolates (C21 and C12) do not correspond to known protein sequences. They are homologous to EST sequences from a fetal brain library. C21 encodes a previously unknown gene that is a member of the WD-40 family. An open reading frame encoding a 515 amino acid protein has been identified. Four mRNAs, differentially expressed by CD34(+)CD38(-) human bone marrow cells, have been identified. Although this population is highly enriched for early hematopoietic progenitors, none of these genes encodes a message whose expression is limited to the hematopoietic system. They all are expressed in a variety of tissues, suggesting that they are involved in processes that are fundamental to the development of many cell types. All of these cDNAs possess atypically long 3' UTRs, and one of them is among the longest ever described. Their differential expression by immature hematopoietic cells, in contrast to more mature cells, suggest that long 3' UTRs may be characteristic of genes that play a regulatory role during development. PMID- 11063879 TI - Erratum PMID- 11063878 TI - Ex vivo expansion of human umbilical cord blood and peripheral blood CD34(+) hematopoietic stem cells. AB - The proliferation and expansion of human hematopoietic stem cells (HSC) in ex vivo culture was examined with the goal of generating a suitable clinical protocol for expanding HSC for patient transplantation.HSC were derived from umbilical cord blood (UCB) and adult patient peripheral blood stem cell collections. HSC were stimulated to proliferate ex vivo by a combination of two growth factors, flt-3 ligand (FL) and thrombopoietin/c-mpl ligand (TPO/ML), and assessed for expansion by flow cytometry.Ex vivo expansion cultures of UCB were maintained for prolonged periods (up to 16 weeks), and sufficient HSC were generated for adult transplantation. In contrast to UCB, FL + TPO/ML did not significantly increase CD34(+) peripheral blood stem cell (PBSC) numbers.UCB-HSC can be expanded in culture to numbers theoretically adequate for safe, rapid engraftment of adult patients. Additional studies are needed to establish the functional activity of expanded UCB-HSC. PMID- 11063880 TI - Maturation of HIV envelope glycoprotein precursors by cellular endoproteases. AB - The entry of enveloped viruses into its host cells is a crucial step for the propagation of viral infection. The envelope glycoprotein complex controls viral tropism and promotes the membrane fusion process. The surface glycoproteins of enveloped viruses are synthesized as inactive precursors and sorted through the constitutive secretory pathway of the infected cells. To be infectious, most of the viruses require viral envelope glycoprotein maturation by host cell endoproteases. In spite of the strong variability of primary sequences observed within different viral envelope glycoproteins, the endoproteolytical cleavage occurs mainly in a highly conserved domain at the carboxy terminus of the basic consensus sequence (Arg-X-Lys/Arg-Arg downward arrow). The same consensus sequence is recognized by the kexin/subtilisin-like serine proteinases (so called convertases) in many cellular substrates such as prohormones, proprotein of receptors, plasma proteins, growth factors and bacterial toxins. Therefore, several groups of investigators have evaluated the implication of convertases in viral envelope glycoprotein cleavage. Using the vaccinia virus overexpression system, furin was first shown to mediate the proteolytic maturation of both human immunodeficiency virus (HIV-1) and influenza virus envelope glycoproteins. In vitro studies demonstrated that purified convertases directly and specifically cleave viral envelope glycoproteins. Although these studies suggested the participation of several enzymes belonging to the convertases family, recent data suggest that other protease families may also participate in the HIV envelope glycoprotein processing. Their role in the physiological maturation process is still hypothetical and the molecular mechanism of the cleavage is not well documented. Crystallization of the hemagglutinin precursor (HA0) of influenza virus allowed further understanding of the molecular interaction between viral precursors and the cellular endoproteases. Furthermore, relationships between differential pathogenicity of influenza strains and their susceptibility to cleavage are molecularly funded. Here we review the most recent data and recent insights demonstrating the crucial role played by this activation step in virus infectivity. We discuss the cellular endoproteases that are implicated in HIV gp160 endoproteolytical maturation into gp120 and gp41. PMID- 11063881 TI - Mutagenic study of the structure, function and biogenesis of the yeast plasma membrane H(+)-ATPase. PMID- 11063883 TI - Dynamics of the mammalian sperm plasma membrane in the process of fertilization. AB - Sexual reproduction requires the fusion of sperm cell and oocyte during fertilization to produce the diploid zygote. In mammals complex changes in the plasma membrane of the sperm cell are involved in this process. Sperm cells have unusual membranes compared to those of somatic cells. After leaving the testes, sperm cells cease plasma membrane lipid and protein synthesis, and vesicle mediated transport. Biophysical studies reveal that lipids and proteins are organized into lateral regions of the sperm head surface. A delicate reorientation and modification of plasma membrane molecules take place in the female tract when sperm cells are activated by so-called capacitation factors. These surface changes enable the sperm cell to bind to the extra cellular matrix of the egg (zona pellucida, ZP). The ZP primes the sperm cell to initiate the acrosome reaction, which is an exocytotic process that makes available the enzymatic machinery required for sperm penetration through the ZP. After complete penetration the sperm cell meets the plasma membrane of the egg cell (oolemma). A specific set of molecules is involved in a disintegrin-integrin type of anchoring of the two gametes which is completed by fusion of the two gamete plasma membranes. The fertilized egg is activated and zygote formation preludes the development of a new living organism. In this review we focus on the involvement of processes that occur at the sperm plasma membrane in the sequence of events that lead to successful fertilization. For this purpose, dynamics in adhesive and fusion properties, molecular composition and architecture of the sperm plasma membrane, as well as membrane derived signalling are reviewed. PMID- 11063882 TI - Structure of lipid bilayers. AB - The quantitative experimental uncertainty in the structure of fully hydrated, biologically relevant, fluid (L(alpha)) phase lipid bilayers has been too large to provide a firm base for applications or for comparison with simulations. Many structural methods are reviewed including modern liquid crystallography of lipid bilayers that deals with the fully developed undulation fluctuations that occur in the L(alpha) phase. These fluctuations degrade the higher order diffraction data in a way that, if unrecognized, leads to erroneous conclusions regarding bilayer structure. Diffraction measurements at high instrumental resolution provide a measure of these fluctuations. In addition to providing better structural determination, this opens a new window on interactions between bilayers, so the experimental determination of interbilayer interaction parameters is reviewed briefly. We introduce a new structural correction based on fluctuations that has not been included in any previous studies. Updated measurements, such as for the area compressibility modulus, are used to provide adjustments to many of the literature values of structural quantities. Since the gel (L(beta)') phase is valuable as a stepping stone for obtaining fluid phase results, a brief review is given of the lower temperature phases. The uncertainty in structural results for lipid bilayers is being reduced and best current values are provided for bilayers of five lipids. PMID- 11063896 TI - The role of serotonin in hot flushes. AB - Hot flushes are experienced in those periods of the female life when estrogen levels are low. Hormone replacement therapy is thus the first choice for treatment of hot flushes. However this treatment is not always accepted or contraindicated for a variety of reasons. Estrogen (and progestogen) strongly interact with a number of neurotransmitters and this has led to a range of non hormonal treatments including compounds that act via the noradrenergic or dopaminergic systems as well as herbal remedies. These treatments (which are shortly reviewed) are not always successful. Surprisingly, apart from treatment with some selective serotonin (5-HT) reuptake inhibitors (SSRI's), up till now, little attention is given to the strong interaction of estrogens with the serotonergic system. These interactions are shortly reviewed. Based on these interactions, a hypothesis on the genesis of hot flushes is postulated. Especially the 5-HT(2A) receptor subtype may play a key role in the occurrence of hot flushes. A number of arguments that support this hypothesis are discussed. PMID- 11063897 TI - Treatment of hot flushes with mirtazapine: four case reports. AB - OBJECTIVE: To evaluate the effect of mirtazapine on the severity of hot flushes and bouts of perspiration in women. METHOD: In two women with depression a reduction in hot flushes was noticed by serendipity during treatment with mirtazapine 15-30 mg/daily. On the basis of this observation clinical studies were extended with two non-depressed and non-anxious women with hot flushes. Both subjects were prescribed mirtazapine daily. RESULTS: Four cases are described as case reports. All subjects reported a practically complete disappearance of hot flushes and associated perspiration, within the first week of treatment. CONCLUSION: Mirtazapine appears to have a substantial ameliorating effect on hot flushes and perspiration bouts. It is postulated that the 5-HT(2A) blocking properties of mirtazapine is accounted in the symptomatic relief of hot flushes. In addition it is hypothesized that the serotonergic system is crucially involved in the pathogenesis of hot flushes and perspiration bouts. Further evaluation in double-blind placebo-controlled studies is encouraged. PMID- 11063898 TI - Immunohistochemical detection of estrogen and progesterone receptor in human cornea. AB - OBJECTIVE: For treatment of postmenopausal keratoconjunctivitis sicca hormone therapy is favored by some clinicians. The likely morphological basis assessing the hormone receptor status in the human cornea has not been performed. Immunohistochemical staining methods provide the opportunity to evaluate the hormone receptor content within the histologic compartments of the cornea. The aim of our study was to assess and localize immunohistochemical hormone receptor staining in the human cornea. METHODS: Formalin-fixed and paraffin-embedded specimens of three pre- and three postmenopausal women were assessed for localization of estrogen receptor (ER) and progesterone receptor (PR) expression with established immunohistochemical hormone receptor staining methods. RESULTS: No nuclear staining reaction was found in the epi- and endothelial layers of the corneas. The stroma of the corneas showed no immunohistochemical staining reaction in all cases. We found cytoplasmatic PR staining of the endothelial layer in two cases. CONCLUSIONS: We found no morphological basis in the human cornea for the use of topical steroid hormone treatment in postmenopausal keratoconjunctivitis sicca. Hormone receptor expression in the conjunctiva or in the lacrimal gland may have an impact in some patients showing relief of symptoms in postmenopausal dry eye syndrome. PMID- 11063899 TI - Gender differences in fracture risk and bone mineral density. AB - We have investigated gender-related differences of bone mineral density and fracture threshold in 136 males (age, 60.7+/-9.3 years) and 337 females (age, 59.7+/-7.8 years) without evidence of secondary osteoporosis. Women and men were examined for total amount of spine fractures and bone mineral density by quantitative computed tomography (QCT) of three non-fractured vertebral bodies L1 L5. Females with lumbar fractures (n=96) when compared with non-fracture women (n=241) were older and had lower bone density at their vertebral sites. Males with vertebral fractures (n=52) were older and had a significantly reduced bone mineral density of the spine when compared with healthy males (n=84). When we compared gender-related vertebral fracture rates, we observed a significantly higher prevalence of vertebral fractures in the male population. After matching males and females for age and bone mineral density to exclude an influence of either variable, we compared the prevalence of vertebral fracture risk in both sexes with logistic regression analysis. Data of estimated fracture risk, differed very significantly for sex and bone density at the vertebral site, indicating that men present fracture at a higher bone density level compared with females; in 10% of study population fractures occurred at a QCT level of 105 mg/cm(3) in women and 120 mg/cm(3) in men. The estimated odds ratio for sex of 3.1 (95% CI) means a three-fold increased risk for vertebral fractures compared to males at a given density level. These results underline that a decreased bone mineral density leads to the occurrence of spine fractures in females and males as well. PMID- 11063900 TI - Hormonal replacement therapy reduces forearm fracture incidence in recent postmenopausal women - results of the Danish Osteoporosis Prevention Study. AB - OBJECTIVES: To study the fracture reducing potential of hormonal replacement therapy (HRT) in recent postmenopausal women in a primary preventive scenario. METHODS: Prospective controlled comprehensive cohort trial: 2016 healthy women aged 45-58 years, from three to 24 months past last menstrual bleeding were recruited from a random sample of the background population. Mean age was 50. 8+/ 2.8 years, and the number of person years followed was 9335.3. There were two main study arms: a randomised arm (randomised to HRT; n=502, or not; n=504) and a non-randomised arm (on HRT; n=221, or not; n=789 by own choice). First line HRT was oral sequential oestradiol/norethisterone in women with intact uterus and oral continuous oestradiol in hysterectomised women. RESULTS: After five years, a total of 156 fractures were sustained by 140 women. There were 51 forearm fractures in 51 women. By intention-to-treat analysis (n=2016), overall fracture risk was borderline statistically significantly reduced (RR=0.73, 95% CI: 0.50 1.05), and forearm fracture risk was significantly reduced (RR=0.45, 95% CI: 0.22 0.90) with HRT. Restricting the analysis to women who had adhered to their initial allocation of either HRT (n=395) or no HRT (n=977) showed a significant reduction in both the overall fracture risk (RR=0.61, 95% CI: 0.39-0.97) and the risk of forearm fractures (RR=0.24, 95% CI: 0.09-0.69). Compliance with HRT was 65% after five years. CONCLUSIONS: It is possible to reduce the number of forearm fractures and possibly the total number of fractures in recent postmenopausal women by use of HRT as primary prevention. PMID- 11063901 TI - Effect of estrogen on intestinal strontium absorption in postmenopausal women. AB - OBJECTIVES: there is considerable uncertainty about the underlying cause of decreased intestinal calcium absorption that occurs in postmenopausal women. In a previous study, estrogen treatment did not result in an increased intestinal calcium absorption using strontium as a marker. A possible explanation could be that the calcium/strontium load given to the women was too high ( approximately 600 mg Ca), which might result in an insensitive test with respect to the possible stimulation of active strontium transport by estrogen. Therefore, the purpose of this study was to reinvestigate the effect of estrogen on active intestinal strontium absorption using a load of 2.5 mmol of strontium only. METHODS: the effect of estrogen on intestinal strontium absorption was measured in eight normal postmenopausal women. The study included two baseline strontium absorption tests, which were performed with an interval of 10 days for calculating the within subject variation (SER). Thereafter the effect of 2 months of estrogen treatment on intestinal strontium absorption was assessed. Fractional absorption (FC(240)) and the area under the concentration time curve (AUC) 4 h after an oral strontium load of 2.5 mmol were calculated. RESULTS: the within subject SER of FC(240) and AUC(0-240) were 2.3+/-0.76 and 1.2+/-0.41, respectively. FC(240) and AUC(0-240) of strontium were unchanged after treatment with estrogen. CONCLUSIONS: in normal postmenopausal women, we did not find a modulating effect of short-term treatment with a (supra) physiological dose of estrogen on intestinal calcium absorption as measured by the strontium absorption test. PMID- 11063902 TI - Plasma leptin levels in obese and non-obese postmenopausal women before and after hormone replacement therapy. AB - OBJECTIVE: the aim of this study was to investigate the effect of hormone replacement therapy (HRT) on plasma leptin levels in postmenopausal women, and the relationship between the plasma leptin levels and obesity. METHODS: premenopausal women with normal cycles (n=30; mean ages, 35.4+/-8.3 years) and postmenopausal women (n=45; mean ages, 49.5+/-4.7 years) were randomly selected. Women were classified as obese (BMI>27 kg/m(2)) and as non-obese (BMI<27 kg/m(2)). Blood samples were obtained from the premenopausal women at the beginning of cycle, and from the postmenopausal women before and 6 months after HRT. Plasma leptin levels were measured by radioimmunassay. RESULTS: plasma leptin levels were significantly higher in premenopausal women than in postmenopausal women (18. 60+/-5.0; 3.67+/-2.44 ng/ml, respectively, P<0.001). Obese premenopausal women (n=15) had significantly higher plasma leptin levels (24. 60+/-7.81 ng/ml) in comparison with the levels of the non-obese premenopausal women (n=15; 12.50+/-4. 63 ng/ml) (P<0.001). Although there was no significant difference in the plasma leptin levels between obese (n=25) and non obese (n=20) postmenopausal women before HRT, plasma leptin levels were significantly elevated in both obese and non-obese postmenopausal women after HRT (P<0.001), and the obese women had significantly higher plasma leptin levels than the non-obese (29.05+/-10.53; 14.78+/-6.76 ng/ml, respectively, P<0.001). CONCLUSION: HRT is effective in the elevation of the plasma leptin levels in postmenopausal women, and in obese women the increase of the plasma leptin levels are more marked than the non-obese women after HRT. PMID- 11063903 TI - Endometrial safety and tolerability of AERODIOL(R) (intranasal estradiol) for 1 year. AB - OBJECTIVE: the purpose of this study was to assess the endometrial safety and patient acceptability of a pulsed estrogen therapy provided by S21400 (intranasal 17 beta-estradiol) in the treatment of postmenopausal symptoms. DESIGN: postmenopausal women (n=408) entered an open-label, community based, multicentre trial. Patients received S21400 plus sequential (>90% of patients) or continuous progestogen. Treatment was initiated with a standard daily dose of 300 microg but dose adaptation was possible every 3 months from 150 to 600 microg daily. Endometrial biopsies were performed at entry and at 12 months, and bleeding patterns were recorded at 3-monthly intervals throughout the trial. RESULTS: 71% of patients received 300 microg per day S21400 throughout the study, 3% had their dose decreased, 19% had their dose increased and 7% had their dose both decreased and increased. Three hundred and eleven biopsies were obtained after 12 months of treatment, there were no cases of endometrial hyperplasia. The 95% confidence interval [CI] for the rate of incidence was 0-1.2%. Cyclical bleeding occurred in 82% of sequential treatment cycles. Unexpected bleeding occurred in 5% of the treatment cycles. Presence of unexpected bleeding varied according to the treatment regimen, 15 and 4% of the cycles with combined continuous and sequential regimen, respectively. Unexpected bleeding was mostly spotting. Nasal treatment was well accepted. Nasal symptoms (itching sensation, rhinorrhea and sneezing) were mostly mild in intensity and they led to treatment withdrawal in approximately 3% of patients. The rate of treatment continuation was 85% at 1 year. CONCLUSIONS: S21400, in combination with continuous or sequential progestogen, exhibits good endometrial safety and patient acceptability in postmenopausal women. PMID- 11063904 TI - Comparison of the long-term effects of oral estriol with the effects of conjugated estrogen on serum lipid profile in early menopausal women. AB - OBJECTIVE: We investigated the long-term effects of oral estriol (E(3)) on serum levels of total cholesterol (t-Cho), high-density lipoprotein cholesterol (HDL Cho), low-density lipoprotein cholesterol (LDL-Cho), and triglycerides in early menopausal women. METHODS: We studied 67 healthy early menopausal women who were treated for 48 months with 2.0 mg of E(3) plus 2.5 mg of medroxyprogesterone acetate daily (E(3) group, n=21), 0.625 mg of conjugated estrogen plus 2.5 mg of medroxyprogesterone acetate daily (CE group, n=19), or 1.0 microg of 1alpha hydroxyvitamin D(3) daily or 1.8 g of calcium lactate containing 250 mg of elemental calcium daily (control group, n=27). The serum levels of t-Cho, HDL Cho, LDL-Cho, and triglycerides were evaluated at baseline and every 6 months. RESULTS: After 48 months of treatment, the t-Cho decreased significantly by 4.3+/ 2.1% (mean+/-SE) from baseline in the E(3) group, did not change in the CE group (-1.9+/-2.1%), and significantly increased (5.4+/-3.4%) in the control group. The HDL-Cho significantly increased in the CE group (10.7+/-2.4%), but not in the E(3) group (3.8+/-3.3%) or in the control group (-3.6+/-3. 0%). The LDL-Cho significantly decreased in the CE group (-11.4+/-4. 0%), did not change in the E(3) group (-5.2+/-3.6%), and significantly increased in the control group (11.8+/-6.3%). The triglyceride level decreased significantly in the E(3) group ( 6. 7+/-4.9%), whereas it significantly increased in the CE group (17. 6+/-11.4%), and did not change in the control group (6.1+/-6.4%). CONCLUSIONS: Oral E(3) prevented a postmenopausal rise in the t-Cho. Oral estriol did not induce the hypertriglyceridemia that was seen after treatment with conjugated estrogen. Oral E(3) may be a useful alternative therapy in women with hypertriglyceridemia and in women who are reluctant to continue conventional hormone replacement therapy because of uterine bleeding. PMID- 11063905 TI - The effect of tibolone versus continuous combined norethisterone acetate and oestradiol on memory, libido and mood of postmenopausal women: a pilot study. AB - OBJECTIVE: Several studies have shown a positive effect of oestrogen on memory, mood and well-being but these data are controversial and focus particularly on the effect of oestrogen alone. In this pilot study we have investigated the effect of a continuous combination of norethisterone acetate 1 mg and oestradiol valerate 2 mg (Kliogest) versus tibolone (Livial) on memory, sexuality and mood. METHODS: Twenty-two postmenopausal women, age range 51-57, were randomised to a 6 months single blind interventional study treatment with either continuous combined oestradiol plus norethisterone acetate, or tibolone. Computerised psychological test of memory, mood and libido were administered both before and at the end of the 6 months treatment. RESULTS: Fourteen patients completed the study; eight on Livial and six on Kliogest. Recognition memory was improved by Kliogest but not by Livial (P<0.05) while either drug equally improved both the reaction time (P<0.01) and accuracy of performance (P<0.001) of categorical semantic memory. Both the treatments improved libido significantly (P<0.05), while the mood did not change with either. CONCLUSION: The results suggest that both these forms of hormonal replacement therapy improve the efficiency of memory performance and libido. However, a combination of oestradiol and norethisterone acetate seems to be marginally more effective on improving cognitive processes. PMID- 11063906 TI - Mitochondrial or cytosolic catalase reverses the MnSOD-dependent inhibition of proliferation by enhancing respiratory chain activity, net ATP production, and decreasing the steady state levels of H(2)O(2). AB - Manganese superoxide dismutase (MnSOD) overexpression has been shown to reverse the malignant phenotype in a variety of tumor cell lines. The inhibition of proliferation and reversal of the malignant phenotype has been attributed to an increase in H(2)O(2) production as a result of the dismutation reaction. However, direct evidence in support of this hypothesis has not been forthcoming. To evaluate the contribution of H(2)O(2) in the regulation of cell growth in response to MnSOD overexpression, control and MnSOD-overexpressing HT-1080 fibrosarcoma cells were transfected with constructs that direct catalase to either the mitochondrial or cytosolic compartments. Overexpression of catalase in either compartment reversed the proliferative and clonogenic inhibition associated with MnSOD overexpression, blocked the increase in the steady state levels of H(2)O(2) as measured by flow cytometric analysis of 2', 7' dichlorofluorescein diacetate, and increased protection from the cytotoxicity of H(2)O(2). In addition, mitochondrial or cytosolic catalase enhances respiration through complex I and II in both control and MnSOD overexpressing cell lines and reverses a MnSOD-dependent decrease in net ATP production. Thus, catalase reverses the proliferative inhibition associated with MnSOD overexpression and may also play an important role in metabolic regulation. PMID- 11063907 TI - Glucose enhancement of LDL oxidation is strictly metal ion dependent. AB - Recent evidence suggests that lipoprotein oxidation is increased in diabetes, however, the mechanism(s) for such observations are not clear. We examined the effect of glucose on low-density lipoprotein (LDL) oxidation using metal ion dependent and -independent oxidation systems. Pathophysiological concentrations of glucose (25 mM) enhanced copper-induced LDL oxidation as determined by conjugated diene formation or relative electrophoretic mobility (REM) on agarose gels. Similarly, iron-induced LDL oxidation was stimulated by glucose resulting in 4- to 6-fold greater REM than control incubations without glucose. In contrast, glucose had no effect on metal ion-independent LDL oxidation by aqueous peroxyl radicals. The effect of glucose on metal ion-dependent LDL oxidation was associated with enhanced reduction of metal ions, and in the case of iron-induced LDL oxidation, was completely inhibited by superoxide dismutase. The effect of glucose was mimicked by other reducing sugars, such as fructose and mannose, and the extent to which each sugar enhanced LDL oxidation was closely linked to its metal ion-reducing activity. Thus, promotion of LDL oxidation by glucose is specific for metal ion-dependent oxidation and involves increased metal ion reduction. These results provide one potential mechanism for enhanced LDL oxidation in diabetes. PMID- 11063908 TI - Evidence for the pro-oxidant effect of gamma-glutamyltranspeptidase-related enzyme. AB - It has been previously reported that the metabolism of reduced glutathione (GSH) by gamma-glutamyltranspeptidase (GGT) in the presence of chelated metals leads to free radical generation and lipid peroxidation (LPO). The present study demonstrates for the first time that an established cell line expressing GGT-rel, a human GGT-related enzyme, metabolizes extracellular GSH to cysteinylglycine (CysGly) in a time-dependent manner when cells were incubated in a medium containing 2.5 mM GSH and 25 mM glycylglycine. Supplementation with 150-165 microM Fe(3+)-EDTA resulted in a reactive oxygen species (ROS) generation process. The resulting data showed a significantly higher level (7.6-fold) of ROS production in the GGT-rel positive cells in comparison with the GGT-rel negative control cells. CysGly and Cys, but not GSH, were responsible for the observed ROS production, as we confirmed by measuring the same process in the presence of Fe(3+)-EDTA and different thiols. A higher iron reduction and an increased LPO level determined by malondialdehyde HPLC measurement were also found in GGT-rel overexpressing cells compared to GGT-rel negative cells. Our data clearly indicate that in the presence of iron, not only GGT, but also GGT-rel has a pro oxidant function by generation of a reactive metabolite (CysGly) and must be taken into account as a potential physiopathological oxidation system. PMID- 11063909 TI - Studies of LDL oxidation following alpha-, gamma-, or delta-tocotrienyl acetate supplementation of hypercholesterolemic humans. AB - In vitro tocotrienols (T3s) have potent vitamin E antioxidant activity, but unlike tocopherols can inhibit cholesterol synthesis by suppressing 3-hydroxy-3 methyl-glutarylCoA (HMG-CoA) reductase. Because hypercholesterolemia is a major risk factor for coronary artery disease and oxidative modification of low-density lipoprotein (LDL) may be involved in atherogenesis, we investigated whether daily supplements of placebo, or alpha-, gamma-, or delta- (alpha-, gamma-, or delta-) tocotrienyl acetates would alter serum cholesterol or LDL oxidative resistance in hypercholesterolemics in a double-blind placebo controlled study. Subjects were randomly assigned to receive placebo (n = 13), alpha- (n = 13), gamma- (n = 12), or delta- (n = 13) tocotrienyl acetate supplements (250 mg/d). All subjects followed a low-fat diet for 4 weeks, then took supplements with dinner for the following 8 weeks while still continuing diet restrictions. Plasma alpha- and gamma-tocopherols were unchanged by supplementation. Plasma T3s were undetectable initially and always in the placebo group. Following supplementation in the respective groups plasma concentrations were: alpha-T3 0.98 +/- 0.80 micromol/l, gamma-T3 0.54 +/- 0.45 micromol/l, and delta-T3 0.09 +/- 0.07 micromol/l. Alpha T3 increased in vitro LDL oxidative resistance (+22%, p <.001) and decreased its rate of oxidation (p <. 01). Neither serum or LDL cholesterol nor apolipoprotein B were significantly decreased by tocotrienyl acetate supplements. This study demonstrates that: (i) tocotrienyl acetate supplements are hydrolyzed, absorbed, and detectable in human plasma; (ii) tocotrienyl acetate supplements do not lower cholesterol in hypercholesterolemic subjects on low-fat diets; and (iii) alpha-T3 may be potent in decreasing LDL oxidizability. PMID- 11063910 TI - Oxidative injury of isolated cardiomyocytes: dependence on free radical species. AB - The contribution of lipid peroxidation to myocardial injury by free radicals (FR) is still unclear. Consequently, we examined the functional damages inflicted on cultured rat cardiomyocytes (CM) during FR stress provoked by the xanthine/xanthine oxidase system (X/XO) or by a hydroperoxidized fatty acid ((9 Z, 11 E, 13 (S), 15 Z)-13-hydroperoxyocta-decatrienoic acid; 13-HpOTrE), in order to simulate in vitro the initial phase and the propagation phase of the FR attack, respectively. Transmembrane potentials were recorded with glass microelectrodes and contractions were monitored photometrically. The EPR spectroscopy showed that X/XO produced superoxide and hydroxyl radicals during 10 min. The X/XO system altered sharply and irreversibly the spontaneous electrical and mechanical activities of the CM. However, the gas chromatographic analysis showed that these drastic functional damages were associated with comparatively moderate membrane PUFA degradation. Moreover, the EPR analysis did not reveal the production of lipid-derived FR. 13-HpOTrE induced a moderate and reversible decrease in electrical parameters, with no change in CM contractions. These results indicate that the functional consequences of FR attack are dependent on the radical species present and do not support the idea that the membrane lipid breakdown is a major factor of myocardial oxidant dysfunction. PMID- 11063911 TI - Exposure of developing oligodendrocytes to cadmium causes HSP72 induction, free radical generation, reduction in glutathione levels, and cell death. AB - Primary cultures of oligodendrocytes were used to study the toxic effects of cadmium chloride. Cell viability was evaluated by the mitochondrial dehydrogenase activity and confirmed by propidium iodide (PI) fluorescence staining. The expression of the 72 kDa stress protein, HSP72, was assayed by Western blot analysis. The results showed that Cd(2+)-induced toxicity was dependent on the time and dose of exposure, as well as on the developmental stage of the cultures. Oligodendrocyte progenitors were more vulnerable to Cd(2+) toxicity than were mature oligodendrocytes. Mature oligodendrocytes accumulated relatively higher levels of Cd(2+) than did progenitors, as determined by (109)CdCl(2) uptake; treatment with the metal ion caused a more pronounced reduction in intracellular glutathione levels and significantly higher free radical accumulation in progenitors. The latter could explain the observed differences in Cd(2+) susceptibility. HSP72 protein expression was increased both in progenitors and in mature cells exposed to Cd(2+). Pretreatment with N-acetylcysteine, a thiocompound with antioxidant activity and a precursor of glutathione, prevented Cd(2+)-induced (i) reduction in glutathione levels and (ii) induction of HSP72 and diminished (i) Cd(2+) uptake and (ii) Cd(2+)-evoked cell death. In contrast, buthionine sulfoximine, an inhibitor of gamma-glutamyl-cysteine synthetase, depleted glutathione, and potentiated the toxic effect of Cd(2+). These results strongly suggest that Cd(2+)-induced cytotoxicity in oligodendrocytes is mediated by reactive oxygen species and is modulated by glutathione levels. PMID- 11063912 TI - The roles played by crucial free radicals like lipid free radicals, nitric oxide, and enzymes NOS and NADPH in CCl(4)-induced acute liver injury of mice. AB - Mice were administered a single dose of carbon tetrachloride (CCl(4)) to induce acute liver injury. We found that lactate dehydrogenase (LDH) and glutamic pyruvic transaminase (GPT) levels in serum, as well as the level of thiobarbituric acid reaction substances (TBARS) in liver homogenate increased significantly in a manner both dose dependent and time dependent after CCl(4) administration. Such results suggest that the liver is susceptible to CCl(4) treatment and that lipid peroxidation is associated with CCl(4)-induced liver injury. The spin-trapping electron paramagnetic resonance (EPR) method was used to detect nitric oxide (NO) level in liver. The chemiluminescence method was also employed to measure the NO(2)(-)/NO(3)(-) concentration in serum. The NO levels in liver tissues and NO(2)(-)/NO(3)(-) concentration in serum were found to decrease significantly both in a dose-dependent manner and in time course after CCl(4) treatment. The nitric oxide synthase (NOS) II activity in the liver, in contrast, was found to increase significantly. Our study suggests that not only should the expression of NOS be analyzed but NO organ and blood concentration must be measured in the study of diseases involving nitric oxide. L-arginine treatment had no significant effect on the liver function of CCl(4)-treated mice. It was found that NO donor sodium nitroprusside (SNP; 50 or 100 microg/kg) treatment resulted in decreases of LDH, GPT, and TBARS levels, leading to a protective effect on CCl(4)-treated mice. On the other hand, N(G)-nitro-L arginine methyl ester (L-NAME, 100 or 300 mg/kg) treatment caused more severe liver damage. Moreover, we have found in an in vitro EPR study that SNP could scavenge lipid peroxyl radical LOO&z.rad;. The above results together suggest that NO may protect CCl(4)-induced liver injury through scavenging lipid radical, inhibiting the lipid peroxidation chain reaction. On the basis of our analysis, we put forth two explanations for the stated discrepancy between NOS II and NO production: (i) NO was used up gradually in terminating lipid peroxidation and (ii) NADPH was depleted (on the basis of correlation evidence only). PMID- 11063913 TI - Deficient iNOS in inflammatory bowel disease intestinal microvascular endothelial cells results in increased leukocyte adhesion. AB - Microvascular endothelial cells play a key role in inflammation by undergoing activation and recruiting circulating immune cells into tissues and foci of inflammation, an early and rate-limiting step in the inflammatory process. We have previously [Binion et al., Gastroenterology112:1898-1907, 1997] shown that human intestinal microvascular endothelial cells (HIMEC) isolated from surgically resected inflammatory bowel disease (IBD) patient tissue demonstrate significantly increased leukocyte binding in vitro compared to normal HIMEC. Our studies [Binion et al., Am. J. Physiol.275 (Gastrointest. Liver Physiol. 38):G592 G603, 1998] have also demonstrated that nitric oxide (NO) production by inducible nitric oxide synthase (iNOS) normally plays a key role in downregulating HIMEC activation and leukocyte adhesion. Using primary cultures of HIMEC derived from normal and IBD patient tissues, we sought to determine whether alterations in iNOS-derived NO production underlies leukocyte hyperadhesion in IBD. Both nonselective (N(G)-monomethyl-L-arginine) and specific (N-Iminoethyl-L-lysine) inhibitors of iNOS significantly increased leukocyte binding by normal HIMEC activated with cytokines and lipopolysaccharide (LPS), but had no effect on leukocyte adhesion by similarly activated IBD HIMEC. When compared to normal HIMEC, IBD endothelial cells had significantly decreased levels of iNOS mRNA, protein, and NO production following activation. Addition of exogenous NO by co culture with normal HIMEC or by pharmacologic delivery with the long-acting NO donor detaNONOate restored a normal leukocyte binding pattern in the IBD HIMEC. These data suggest that loss of iNOS expression is a feature of chronically inflamed microvascular endothelial cells, which leads to enhanced leukocyte binding, potentially contributing to chronic, destructive inflammation in IBD. PMID- 11063914 TI - Inhibition of vascular NADH/NADPH oxidase activity by thiol reagents: lack of correlation with cellular glutathione redox status. AB - Vascular NAD(P)H oxidase activity contributes to oxidative stress. Thiol oxidants inhibit leukocyte NADPH oxidase. To assess the role of reactive thiols on vascular oxidase, rabbit iliac/carotid artery homogenates were incubated with distinct thiol reagents. NAD(P)H-driven enzyme activity, assessed by lucigenin (5 or 250 microM) luminescence, was nearly completely (> 97%) inhibited by the oxidant diamide (1mM) or the alkylator p-chloromercuryphenylsulfonate (pCMPS, 0.5mM). Analogous inhibition was also shown with EPR spectroscopy using DMPO as a spin trap. The oxidant dithionitrobenzoic acid (0.5mM) inhibited NADPH-driven signals by 92% but had no effect on NADH-driven signals. In contrast, the vicinal dithiol ligand phenylarsine oxide (PAO, 1 microM) induced minor nonsignificant inhibition of NADPH-driven activity, but significant stimulation of NADH triggered signals. The alkylator N-ethyl maleimide (NEM, 0.5mM) or glutathione disulfide (GSSG, 3mM) had no effect with each substrate. Coincubation of N acetylcysteine (NAC, 3mM) with diamide or pCMPS reversed their inhibitory effects by 30-60%, whereas NAC alone inhibited the oxidase by 52%. Incubation of intact arterial rings with the above reagents disclosed similar results, except that PAO became inhibitor and NAC stimulator of NADH-driven signals. Notably, the cell impermeant reagent pCMPS was also inhibitory in whole rings, suggesting that reactive thiol(s) affecting oxidase activity are highly accessible. Since lack of oxidase inhibition by NEM or GSSG occurred despite significant cellular glutathione depletion, change in intracellular redox status is not sufficient to account for oxidase inhibition. Moreover, the observed differences between NADPH and NADH-driven oxidase activity point to complex or multiple enzyme forms. PMID- 11063915 TI - Antioxidant properties of di-tert-butylhydroxylated flavonoids. AB - Epidemiological evidence suggests an inverse relationship between dietary intake of flavonoids and cardiovascular risk. The biological activities of flavonoids are related to their antioxidative effects, but they also can be mutagenic, due to the prooxidant activity of the catechol pattern. To prevent these problems, we synthesized new flavonoids where one or two di-tert-butylhydroxyphenyl (DBHP) groups replaced catechol moiety at position 2 of the benzopyrane heterocycle. Two DBHP moieties can also be arranged in an arylidene structure or one DBHP fixed on a chalcone structure. Position 7 on the flavone and arylidene or position 4 on the chalcone was substituted by H, OCH(3), or OH. New structures were compared with quercetin and BHT in an LDL oxidation system induced by Cu(II) ions. Arylidenes and chalcones had the best activities (ED(50) = 0.86 and 0.21) compared with vitamin E, BHT, and quercetin (ED(50) = 10.0, 7. 4, and 2.3 microM). Activity towards stable free radical 1, 1-diphenyl-2-picryl-hydrazyl (DPPH) was measured by log Z and ECR(50) parameters. Synthesized flavones proved to be poor DPPH radical scavengers, the activity increasing with the number of DBHP units. In contrast, arylidenes and chalcones were stronger DPPH radical scavengers (log Z > 3, 0.3 < ECR(50) < 2.12) than BHT (log Z = 0.75, ECR(50) = 12.56) or quercetin (log Z = 2.76, ECR(50) = 0.43). Unlike quercetin, synthesized compounds neither chelated nor reduced copper, proving that these new flavonoids had no prooxidant activity in vitro. PMID- 11063916 TI - Mitochondrial glutathione depletion by glutamine in growing tumor cells. AB - The effect of L-glutamine (Gln) on mitochondrial glutathione (mtGSH) levels in tumor cells was studied in vivo in Ehrlich ascites tumor (EAT)-bearing mice. Tumor growth was similar in mice fed a Gln-enriched diet (GED; where 30% of the total dietary nitrogen was from Gln) or a nutritionally complete elemental diet (SD). As compared with non-tumor-bearing mice, tumor growth caused a decrease of blood Gln levels in mice fed an SD but not in those fed a GED. Tumor cells in mice fed a GED showed higher glutaminase and lower Gln synthetase activities than did cells isolated from mice fed an SD. Cytosolic glutamate concentration was 2 fold higher in tumor cells from mice fed a GED ( approximately 4 mM) than in those fed an SD. This increase in glutamate content inhibited GSH uptake by tumor mitochondria and led to a selective depletion of mitochondrial GSH (mtGSH) content (not found in mitochondria of normal cells such as lymphocytes or hepatocytes) to approximately 57% of the level found in tumor mitochondria of mice fed an SD. In tumor cells of mice fed a GED, 6-diazo-5-norleucine- or L glutamate-gamma-hydrazine-induced inhibition of glutaminase activity decreased cytosolic glutamate content and restored GSH uptake by mitochondria to the rate found in EAT cells of mice fed an SD. The partial loss of mtGSH elicited by Gln did not affect generation of reactive oxygen intermediates (ROIs) or mitochondrial functions (e.g., intracellular peroxide levels, O(2)(-)(*) generation, mitochondrial membrane potential, mitochondrial size, adenosine triphosphate and adenosine diphosphate contents, and oxygen consumption were found similar in tumor cells isolated from mice fed an SD or a GED); however, mitochondrial production ROIs upon TNF-alpha stimulation was increased. Our results demonstrate that glutamate derived from glutamine promotes an inhibition of GSH transport into mitochondria, which may render tumor cells more susceptible to oxidative stress-induced mediators. PMID- 11063917 TI - Clinical and physiological consequences of rapid tryptophan depletion. AB - We review here the rapid tryptophan depletion (RTD) methodology and its controversial association with depressive relapse. RTD has been used over the past decade to deplete serotonin (5-hydroxy-tryptamine, or 5-HT) in humans and to probe the role of the central serotonin system in a variety of psychiatric conditions. Its current popularity was stimulated by reports that RTD reversed the antidepressant effects of selective serotonin reuptake inhibitors (SSRIs) and monoamine oxidase inhibitors (MAOIs) in remitted patients with a history of depression but not in patients treated with antidepressants which promote catecholaminergic rather than serotonergic neurotransmission (such as tricyclic antidepressants or buproprion). However, RTD has inconsistent effects in terms of full clinical relapse in depressed patients. Pooling the data from all published reports, patients who are either unmedicated and/or fully remitted are much less likely to experience relapse (7 of 61, or approximately 9%) than patients who are recently medicated and partially remitted (63 of 133, or approximately 47%; although, the numbers here may reflect patient overlap between reports). Recently remitted patients who have been treated with non-pharmacological therapies such as total sleep deprivation, electroconvulsive therapy, or bright light therapy also do not commonly show full clinical relapse with RTD. We briefly review RTD effects in other psychiatric disorders, many of which are treated with SSRIs. There is accumulating evidence to suggest that RTD affects central serotonergic neurotransmission. Nevertheless, many questions remain about the ability of RTD to reverse the beneficial effects of SSRIs or MAOIs, or to induce symptoms in unmedicated symptomatic or asymptomatic patients. PMID- 11063918 TI - Regulation of sensorimotor gating of the startle reflex by serotonin 2A receptors. Ontogeny and strain differences. AB - Sensorimotor gating of the startle reflex can be assessed via measures of prepulse inhibition (PPI), which is the reduction in startle magnitude when the startling stimulus is preceded immediately by a weak prepulse. PPI is reduced in humans with specific neuropsychiatric disorders and in rats after treatment with certain classes of drugs, including serotonin (5-HT) agonists. Because of the relative loss of PPI in inherited, neurodevelopmental disorders such as schizophrenia, there is great interest in understanding the inherited and developmental features of the neurochemical regulation of PPI in animals. In the present study, PPI was disrupted significantly by the 5-HT2A agonist 2, 5 dimethoxy-4 iodopheny-lisopropylamine (DOI) in Sprague Dawley (SDH) and Wistar rat strains (WH). While it was demonstrated that the DOI effects in SDH rats reflected an unequivocal disruption of sensorimotor gating, in WH rats, reduced PPI was observed in the context of a trend for a DOI-induced reduction in startle magnitude. This effect of DOI in SDH rats was evident at the earliest date tested (17 days of age) in male pups, but was not statistically significant in female pups. Thus, the regulation of sensorimotor gating by 5-HT2A receptor stimulation in rats may exhibit subtle differences across strains, and within SDH rats, between sexes. Most importantly, the 5-HT2A regulation of sensorimotor gating in male SDH rats is a "phenotype" that is expressed very early in life, and is sustained through adulthood. PMID- 11063919 TI - Involvement of the nucleus accumbens and medial prefrontal cortex in the expression of conditioned hyperactivity to a cocaine-associated environment in rats. AB - This study examined the roles of the nucleus accumbens (NAc), medial prefrontal cortex, basolateral amygdala, and ventral subiculum of the hippocampus in the expression of Pavlovian conditioned hyperactivity responses to cocaine-related stimuli. This was accomplished by pharmacologically inhibiting these regions prior to drug-free tests for conditioned hyperactivity in an environment previously associated with cocaine. The results indicate that conditioned hyperactivity could be disrupted by infusions of the GABA-B agonist, baclofen (0.2 nmol/0.5 microl/side) into the NAc, or completely blocked by infusions of the GABA-A agonist, muscimol (0.1 and 0.2 nmol/0.5 microl/side) into the medial prefrontal cortex. In contrast, conditioned hyperactivity was unaffected by pharmacological inhibition of the basolateral amygdala, the ventral subiculum, or sites dorsal to the NAc or prefrontal cortex. These findings suggest that the NAc and the prefrontal cortex are crucial elements of the neural circuitry underlying the expression of Pavlovian conditioned responses to cocaine-related stimuli. PMID- 11063920 TI - Ketamine effects on eye movements. AB - In order to determine if the N-methyl-D-aspartate antagonist ketamine would reproduce eye movement dysfunction in schizophrenia, we studied 12 normal control subjects with low dose (0.1 mg/kg) bolus injection of ketamine in a double-blind placebo-controlled study. Oculomotor measures were obtained during smooth pursuit that included closed loop gain and measures of gain during masking conditions. Measures during initiation of smooth pursuit included latency, open loop acceleration and velocity. Ketamine disrupted closed loop gain and open loop acceleration but not measures during the masking conditions. The ketamine partly reproduced some abnormalities seen in schizophrenia but not measures that may be more specifically linked to familial abnormalities found in family members of subjects with schizophrenia. PMID- 11063921 TI - Birth insult alters dopamine-mediated behavior in a precocial species, the guinea pig. Implications for schizophrenia. AB - Schizophrenia is associated with increased birth complications, suggesting that birth complications might alter CNS dopaminergic activity later in life. In rats, Caesarean section (C-section) birth can produce long term changes in dopaminergic biochemistry and behavior. However rat brain is somewhat immature compared to human brain at birth. The current study tested if mild birth complications also alter dopamine-mediated function in a species with a more mature CNS at birth, the guinea pig. As adults, guinea pigs born by C-section showed increased amphetamine-induced locomotion and disruption of prepulse inhibition (PPI) of acoustic startle, compared to vaginally born controls. Guinea pigs born by C section with 1 min of added global anoxia showed reduced amphetamine-induced locomotion and disrupted PPI, while a C-section plus 2 min anoxia group showed no change in amphetamine-induced locomotion but increased amphetamine-induced startle. No group differences in effects of amphetamine or apomorphine on PPI were observed. Taken with previous findings, these results indicate that mild birth complications can cause long term changes in dopamine-mediated behavior in both guinea pig and rat, two species spanning the level of human brain maturity at birth. PMID- 11063922 TI - In vivo measurement of the vesicular monoamine transporter in schizophrenia. AB - Given evidence for excessive striatal dopamine activity in schizophrenia, we sought to test the hypothesis that dopaminergic innervation in the striatum is abnormally elevated, and a secondary hypothesis that age-related loss is accelerated. Twelve schizophrenic subjects on stable doses of medications, along with 12 age and sex-matched healthy control subjects, underwent positron emission tomography (PET) studies with [11C]dihydrotetrabenazine (DTBZ), which binds to the vesicular monoamine transporter, type 2 (VMAT2). DTBZ binding reflects principally dopaminergic projections in the striatum and appears in animal models, over treatment periods as long as two weeks, not to be regulated by antipsychotic drugs. Using an equilibrium analysis, we obtained measurements of the binding potential (BP) of [11C]DTBZ, as well as a transport (K(1)) measure, corresponding to regional cerebral blood flow. BP in the striatum showed no difference between the patient and control groups, and no differential effect of age. We did not find evidence supporting the hypothesis that excessive dopamine activity in schizophrenia could be explained by increased density of striatal dopamine terminals. PMID- 11063923 TI - Characterization of inhibition by risperidone of the inwardly rectifying K(+) current in pituitary GH(3) cells. AB - The effects of risperidone on ionic currents in rat pituitary GH(3) cells were investigated with the aid of the patch-clamp technique. Hyperpolarization activated K(+) currents in GH(3) cells bathed in high-K(+) Ca(2+)-free solution were studied to determine the effect of risperidone and other related compounds on the inwardly rectifying K(+) current (I(K(IR))). Risperidone (0.1-10 microM) suppressed the amplitude of I(K(IR)) in a concentration-dependent manner. The IC(50) value for the risperidone-induced inhibition of I(K(IR)) was 1 microM. Risperidone (3 microM) was found to slow the rate of activation. An increase in current deactivation by the presence of risperidone was also observed. Haloperidol (10 microM) and thioridazine (10 microM) inhibited the amplitude of I(K(IR)) effectively, and clozapine slightly suppressed it; however, metoclopramide (10 microM) had no effect on it. Risperidone (10 microM) had no effect on voltage-dependent K(+) and L-type Ca(2+) currents. However, in the inside-out configuration, risperidone (10 microM) did not alter the single channel conductance, but reduced the activity of large-conductance Ca(2+) activated K(+) (BK(Ca)) channels. Under the current-clamp mode, risperidone (3 microM) depolarized the membrane potential and increased the firing rate. With the aid of the spectral analysis, cells that exhibited an irregular firing pattern were also converted to those displaying a regular firing pattern after addition of risperidone (3 microM). The present study provides evidence that risperidone, in addition to the blockade of dopamine receptors, can produce a depressant effect on I(K(IR)) and BK(Ca) channels, and implies that the blockade of these ionic currents by risperidone may affect membrane excitability and prolactin secretion in GH(3) cells. PMID- 11063924 TI - Treatment response in depressed patients with enhanced Ca mobilization stimulated by serotonin. AB - Serotonin (5-HT)-stimulated intraplatelet calcium (Ca) mobilization has been shown to be enhanced in nonmedicated depressive patients by many studies. However, there has not been any longitudinal follow-up study of this parameter. We examined the relationship between treatment response and pretreatment value of the 5-HT-induced Ca response. The 5-HT(10 microM)-induced intraplatelet Ca mobilization was measured in 98 nonmedicated depressive patients (24 bipolar disorders, 51 melancholic major depressive disorders, and 23 non-melancholic major depressive disorders). These patients were followed up prospectively for a further period of five years. The depressed patients with enhanced Ca response to 5-HT in bipolar disorders exhibited a good response to mood stabilizers but those with major depressive disorders showed a poor response to antidepressants. These findings suggest the possibility that the 5-HT-induced intraplatelet Ca response may be a good predictor of treatment response in depressed patients. Longer longitudinal follow-up studies are needed in larger samples to examine if this parameter may be a specific biological marker for unipolar-bipolar dichotomy. PMID- 11063925 TI - Autoradiographic comparison of [3H]-clonidine binding to non-adrenergic sites and alpha(2)-adrenergic receptors in human brain. AB - Clonidine is a partial agonist at brain alpha(2)-adrenoceptors (alpha(2)AR), but also has high affinity (K(D) = 51 nM) in homogenate binding assays for non adrenergic imidazoline-binding sites (I-sites; imidazoline receptors). Herein, an autoradiographic comparison of [3H]-clonidine binding to I-sites and alpha(2)AR in sections of human brain is reported. For I-sites, the adrenergic component of 50 nM [3H]-clonidine binding was masked with either 60 microM norepinephrine (NE; alpha(2)AR agonist) or 12.5 microM methoxy-idazoxan (MIDX; selective alpha(2)AR antagonist), whereas the remaining non-adrenergic sites were studied by displacement with 20 microM cirazoline. Levels of [3H]-clonidine binding to alpha(2)AR and I-sites, determined in adjacent tissue sections, were positively correlated across 27 brain regions (p = 0.0003; r(2) = 0.385). The principal olivary nucleus and the rostral portion of the ventrolateral medulla had highest ratios of I-sites: alpha(2)AR (>4:1). Quantitative transepts drawn across hippocampal images revealed alpha(2)AR enrichments in the CA-1 and inner molecular layers of the dentate gyrus-areas not enriched in I-sites. Competition curves were generated for I-sites in caudate sections using 10 ligands known to distinguish between I(1) and I(2) subtypes. The rank-order of affinities were cirazoline > harmane > BDF6143 > idazoxan = tizanidine (affinities of agmatine, efaroxan, moxonidine, NE, and oxymetazoline were too low to be reliable). Only the endogenous I-site ligand, harmane, had a monophasic displacement curve at the non-adrenergic sites (Ki = 521 +/- 12 nM). IN CONCLUSION: 1) the distribution of non-adrenergic [3H]-clonidine binding sites in human brain sections was correlated with, but distinct from alpha(2)AR; and 2) the affinities of these sites was distinct from alpha(1)AR, alpha(2)AR, I(1) or I(2) sites as previously defined in membrane binding assays. The properties of this non-adrenergic [3H] clonidine binding site are consistent with I-sites previously labeled by [3H] cirazoline in rat brain. PMID- 11063927 TI - Web alert AB - A selection of World Wide Web sites relevant to papers published in this issue of Current Opinion in Cell Biology. PMID- 11063926 TI - Alterations in endogenous brain beta-endorphin release by adrenal medullary transplants in the spinal cord. AB - While transplants of adrenal medullary cells into the spinal subarachnoid space may produce antinociception via inhibition of spinal pain transmission pathways, alterations at higher central nervous system (CNS) centers have not been addressed. Recent findings suggest that prolonged noxious stimulation results in release of endogenous beta-endorphin in the brain, possibly as a compensatory mechanism to reduce nociception. The goal of this study was to determine whether adrenal medullary transplants in the spinal subarachnoid space alter endogenous beta-endorphin secretion in the hypothalamic arcuate nucleus, its principal CNS source. Pain behaviors and arcuate beta-endorphin secretion by microdialysis were monitored during the formalin pain test in animals with spinal adrenal medullary or control transplants. Basal levels of extracellular beta-endorphin were 3-fold higher in adrenal medullary-implanted than in controls. In control animals, formalin induced robust pain behaviors and a marked transient increase in beta endorphin release 30-60 min following injection. In contrast, pain behaviors were attenuated and the formalin-induced increase in beta-endorphin was completely blocked in adrenal medullary implanted animals. Findings from these studies indicate that adrenal medullary transplants in the spinal subarachnoid space can alter beta-endorphin release in the arcuate nucleus both basally and in response to noxious stimuli. Thus, spinally placed adrenal medullary transplants not only alter local spinal cord pharmacology, but can alter endogenous neurochemistry at higher pain processing centers as well. PMID- 11063928 TI - Editorial overview PMID- 11063929 TI - Control of mitosis by changes in the subcellular location of cyclin-B1-Cdk1 and Cdc25C. AB - Nuclear events of mitosis are initiated when the protein kinase cyclin-B1-Cdk1 is translocated into the nucleus during prophase. Recent work has unveiled many of the mechanisms that govern the localization of cyclin-B1-Cdk1 and its regulator Cdc25C. Phosphorylation-dependent changes in the rate of nuclear import and export of these proteins help to control the onset of mitosis both in normal cells and in cells delayed before mitosis by DNA damage. PMID- 11063930 TI - Regulation of the meiotic cell cycle in oocytes. AB - The mitotic and meiotic cell cycle share many regulators, but there are also important differences between the two processes. The meiotic maturation of Xenopus oocytes has proved useful for understanding the regulation of Cdc2-cyclin B, a key activator of G2/M progression. New insights have been made recently into the signalling mechanisms that induce G2-arrested oocytes to resume and complete the meiotic cell cycle. PMID- 11063931 TI - Regulation of G(1) cyclin-dependent kinases in the mammalian cell cycle. AB - Cyclin-dependent kinases are the key regulators of cell-cycle transitions. In mammalian cells, Cdk2, Cdk4, Cdk6 and associated cyclins control the G(1) to S phase transition. Because proper regulation of this transition is critical for an organism's survival, these protein kinases are exquisitely regulated at different mechanistic levels and in response to a large variety of intrinsic and extrinsic signals. PMID- 11063932 TI - Chromatin remodeling and Rb activity. AB - Progression of cells through the cell cycle is central to normal cell proliferation, and checkpoints that regulate this cycle are targets of tumorigenic mutations. One of these checkpoints is the Rb family of proteins that seems to regulate exit of cells from both G(1) and S phase of the cell cycle. Recent studies have linked the function of the Rb family to chromatin remodeling enzymes. PMID- 11063933 TI - Eukaryotic DNA replication: from pre-replication complex to initiation complex. AB - A common mechanism has emerged for the control of the initiation of eukaryotic DNA replication. The minichromosome maintenance protein complex (MCM) and Cdc45 have now been recognized as central components of the initiation machinery. In addition, two types of S phase promoting kinases conserved between yeast and humans play critical roles in the initiation reaction. At the onset of S phase, S phase kinases promote the association of Cdc45 with MCM at origins. Upon the formation of the MCM-Cdc45 complex at origins, the duplex DNA is unwound and various replication proteins, including DNA polymerases, are recruited onto unwound DNA. The increasing number of newly identified factors involved in the initiation reaction indicates that the control of initiation requires highly evolved machinery in eukaryotic cells. PMID- 11063934 TI - Cell-cycle checkpoint kinases: checking in on the cell cycle. AB - In response to DNA damage, cell-cycle checkpoints integrate cell-cycle control with DNA repair. The idea that checkpoint controls are an integral component of normal cell-cycle progression has arisen as a result of studies in Drosophila and mice. In addition, an appreciation that DNA damage arises as a natural consequence of cellular metabolism, including DNA replication itself, has influenced thinking regarding the nature of checkpoint pathways. PMID- 11063935 TI - Genes involved in senescence and immortalization. AB - Senescence is now understood to be the final phenotypic state adopted by a cell in response to several distinct cell physiological processes, including proliferation, oncogene activation and oxygen free radical toxicity. The role of telomere maintenance in immortalization and the roles of p16(INK4A), p19(ARF), p53 and other genes in senescence are being further elucidated. Significant progress continues to be made in our understanding of cellular senescence and immortalization. PMID- 11063936 TI - Microarrays and cell cycle transcription in yeast. AB - Microarrays have been used to characterize gene expression through the yeast cell cycle. Computational methods have been applied to the microarray data to identify coregulated clusters of genes, and motif-finding algorithms have found promoter elements characteristic of each cluster. The functional relevance of these promoter elements can be tested using chromatin immunoprecipitation, additional microarrays and other molecular techniques. The yeast forkhead proteins have been successfully identified as cell cycle transcription factors for an important cluster of genes by this and other approaches. PMID- 11063937 TI - Editorial overview PMID- 11063938 TI - Molecular control of neural crest formation, migration and differentiation. AB - Induction, migration and differentiation of the neural crest are crucial for the development of the vertebrate embryo, and elucidation of the underlying mechanisms remains an important challenge. In the past year, a novel signal regulating the formation of neural crest cells has been identified, and advances have been made in uncovering roles for bone morphogenetic protein signals and for a transcription factor in the onset of neural crest migration. There have been new insights into the migration and plasticity of branchial neural crest cells. Important progress has been made in dissecting the roles of bone morphogenetic protein, Wnt and Notch signalling systems and their associated downstream transcription factors in the control of neural crest cell differentiation. PMID- 11063939 TI - Genes that control the development of migrating muscle precursor cells. AB - Skeletal muscles in vertebrates, despite their functional and biochemical similarities, are generated via diverse developmental mechanisms. A major subclass of hypaxial muscle groups is derived from long-range migrating progenitor cells that delaminate from the dermomyotome. The development of this lineage is controlled by Pax3, the c-Met tyrosine kinase receptor, its ligand SF/HGF (scatter factor/hepatocyte growth factor) and the homeobox factor Lbx1. These molecules are essential for establishment of the precursor pool, delamination, migration and target finding. Progress has been made in understanding patterning of the muscles, which requires a precise control of proliferation and differentiation of myogenic precursor cells. PMID- 11063940 TI - Genetic control of branching morphogenesis during Drosophila tracheal development. AB - Branching morphogenesis is a widely used strategy to increase the surface area of a given organ. A number of tissues undergo branching morphogenesis during development, including the lung, kidney, vascular system and numerous glands. Until recently, very little has been known about the genetic principles underlying the branching process and about the molecules participating in organ specification and branch formation. The tracheal system of insects represents one of the best-characterised branched organs. The tracheal network provides air to most tissues and its development during embryogenesis has been studied intensively at the morphological and genetic level. More than 30 genes have been identified and ordered into sequential steps controlling branching morphogenesis. These studies have revealed a number of important principles that might be conserved in other systems. PMID- 11063941 TI - Otx2, Gbx2 and Fgf8 interact to position and maintain a mid-hindbrain organizer. AB - A decade ago, chick-quail transplantation studies demonstrated that the junction between the midbrain and hindbrain has the properties of an organizing center capable of patterning the midbrain and cerebellum. Many of the genes that function to pattern these tissues have been identified and extensively studied. Recent experiments have shown that Otx2, Gbx2 and Fgf8 genes play a major role in the positioning and functioning of this organizing center. PMID- 11063942 TI - From the bottom of the heart: anteroposterior decisions in cardiac muscle differentiation. AB - Recently, studies on specification of axes in the developing embryo have focused on the heart, which is the first functional organ to form and probably responds to common cues controlling positional information in surrounding tissues. The early differentiation of heart cells affords an opportunity to link the acquisition of regional identity with the signals underlying terminal differentiation. In the past year, a wealth of information on these signals has emerged, elucidating the general pathways controlling body axes in the context of the developing heart. PMID- 11063943 TI - Vertebrate segmentation: is cycling the rule? AB - Vertebrate segmentation initiates with the subdivision of the paraxial mesoderm into a regular array of somites. Recent evidence suggests that the segmentation clock - a biochemical oscillator acting in the unsegmented paraxial mesoderm cells in most vertebrates - controls cyclic Notch signalling, resulting in periodic formation of somite boundaries. PMID- 11063944 TI - The nucleolus: the magician's hat for cell cycle tricks PMID- 11063945 TI - The time has come to promote true day surgery. PMID- 11063946 TI - Laboratory tests in children undergoing ambulatory surgery: a review of clinical practice and scientific studies. AB - The drive for cost containment in the United States has lead anesthesiologists to re-assess the benefits of routine pre-operative laboratory and radiological testing. The value of routine tests has been questioned not only by insurance companies but also by physicians. Common pre-operative laboratory and radiological tests are reviewed in the following analysis. Specifically, the use of such tests in children scheduled for ambulatory surgery is discussed. Current clinical practice patterns of pediatric anesthesiologists are included so that physicians may make conclusions on the basis of published literature and clinical practice of peers. PMID- 11063947 TI - Anterior cruciate ligament reconstruction as a day case with extended recovery. AB - The aim of this study was to describe the procedures and the postoperative outcome of arthroscopic anterior cruciate ligament (ACL) reconstruction when carried out a day case with extended recovery. Between December 1995 and September 1998, 91 patients underwent surgery using bone-patellatendon-bone autografts and interference screw fixation. Additional surgical procedures were performed on 35 of the patients. The patient records were evaluated for a mean of 17 months (1-33 months) postoperatively. The course of treatment was. (1) Evaluation and KLT-arthrometer test 14 days preoperatively. (2) Surgery, cryocuff, bupivacain, paracetamol, NSAID and ketobemidon for postoperative pain control. (3) Discharge from hospital within 24 h. (4) Physiotherapy after 14 days. (5) Follow-up after 6 weeks with bandage removal and after 6 months. Eight patients required one further day of hospitalisation due to pain (four), nausea (one), haematoma (two) and prolonged anaesthesia (one). Five patients were readmitted to hospital for a mean of 8 (3-16) days postoperatively. Three patients underwent re-surgery due to haematoma/rupture of the scar. No deep infections were found. We concluded that this effective method of ACL reconstruction can be carried out safely as a day case procedure with extended recovery to the benefit of the patients. PMID- 11063948 TI - Anesthesia and surgical repair of aponeurotic hernias in ambulatory surgery. AB - The aim of this study was to evaluate our 5 year experience in the surgery of umbilical (UH) and epigastric hernias (EH) on an ambulatory basis. Sixty three point seven of UH (88/138) and 68.4% of EH (13/19) could be successfully operated in our ambulatory unit. Morbid obesity, ASA III-IV and insulin dependent diabetes were exclusion criteria. After a preoperative local anesthesia infiltration with 1% lidocaine a repair was undertaken in all 101 patients under monitored anesthesia care. Most patients underwent a mesh hernioplasty as definite treatment. Only three patients could not be discharged on the day of operation. There has been a 2% recurrence rate in long term follow-up. These results demonstrate that two thirds of primary aponeurotic hernias can be satisfactorily operated on ambulatory basis. PMID- 11063949 TI - Fees for outpatient operations in Germany: development, evaluation and European comparison. AB - In Germany, discussions on the fees for statutory sickness insurance for ambulatory surgery has, in the last few years, become almost a symbol of dispute for the German health services. Outpatient surgeons complain about the fact that the fees do not cover their services. They see innovation severely threatened by bureaucracy, profitability by planned economy, rights by reasons of State, aggravated by the 'reform' attempts of the Greens and Socialist coalition Federal Government. On the other hand their opponents complain about the money mindedness of the doctors. Intentional panic or real disaster? The fundamental consideration to clarify this question is based on a comparison of the German statutory medical insurance fees and private fees with our neighbours. In Europe an economic area with similar prices for goods, services and wages, even 'outpatient operations' services with comparable cost rates should be paid for at a corresponding level. Any discrepancies would give cause to look for an explanation by analysing the historic development of fees and the question of a fair comparison between operations and the non-operative services. PMID- 11063950 TI - Fast tracking in ambulatory surgery. AB - Fast tracking after ambulatory surgery is a new paradigm which involves transferring patients from the operating room to the phase II recovery unit (i.e. bypassing the postanaesthesia care unit). The success of fast tracking depends upon appropriate modification of the anaesthetic technique, which would allow rapid emergence from anaesthesia, and the prevention of common postoperative complications such as pain, nausea and vomiting using a multimodal approach. Implementation of a fast track program involves use of clinical pathways that would reduce hospital stay and ensure patient safety. Finally, the concept of fast tracking should be expanded to the overall postoperative recovery, not just bypassing the postanaesthesia care unit. PMID- 11063951 TI - How much ambulatory surgery in the World in 1996-1997 and trends? PMID- 11063952 TI - A comparison of cilostazol and pentoxifylline for treating intermittent claudication. AB - PURPOSE: We performed a randomized, double-blind, placebo-controlled, multicenter trial to evaluate the relative efficacy and safety of cilostazol and pentoxifylline. PATIENTS AND METHODS: We enrolled patients with moderate-to severe claudication from 54 outpatient vascular clinics, including sites at Air Force, Veterans Affairs, tertiary care, and university medical centers in the United States. Of 922 consenting patients, 698 met the inclusion criteria and were randomly assigned to blinded treatment with either cilostazol (100 mg orally twice a day), pentoxifylline (400 mg orally 3 times a day), or placebo. We measured maximal walking distance with constant-speed, variable-grade treadmill testing at baseline and at 4, 8, 12, 16, 20, and 24 weeks. RESULTS: Mean maximal walking distance of cilostazol-treated patients (n = 227) was significantly greater at every postbaseline visit compared with patients who received pentoxifylline (n = 232) or placebo (n = 239). After 24 weeks of treatment, mean maximal walking distance increased by a mean of 107 m (a mean percent increase of 54% from baseline) in the cilostazol group, significantly more than the 64-m improvement (a 30% mean percent increase) with pentoxifylline (P <0.001). The improvement with pentoxifylline was similar (P = 0.82) to that in the placebo group (65 m, a 34% mean percent increase). Deaths and serious adverse event rates were similar in each group. Side effects (including headache, palpitations, and diarrhea) were more common in the cilostazol-treated patients, but withdrawal rates were similar in the cilostazol (16%) and pentoxifylline (19%) groups. CONCLUSION: Cilostazol was significantly better than pentoxifylline or placebo for increasing walking distances in patients with intermittent claudication, but was associated with a greater frequency of minor side effects. Pentoxifylline and placebo had similar effects. PMID- 11063953 TI - Acute infectious mononucleosis: characteristics of patients who report failure to recover. AB - PURPOSE: We sought to determine how often acute mononucleosis precipitates chronic illness, and to describe the demographic, clinical, and psychosocial features that characterize patients who report failure to recover. SUBJECTS AND METHODS: We enrolled 150 patients with infectious mononucleosis during the acute illness and asked them to assess their recovery at 2 and 6 months. At baseline, we performed physical and laboratory examinations; obtained measures of psychological and somatic functioning, social support, and life events; and administered a structured psychiatric interview. RESULTS: Self-assessed failure to recover was reported by 38% of patients (55 of 144) at 2 months and by 12% (17 of 142) at 6 months. Those who had not recovered reported a persistent illness characterized by fatigue and poor functional status. No objective measures of disease, including physical examination findings or serologic or laboratory markers, distinguished patients who failed to recover from those who reported recovery. Baseline predictors for failure to recover at 2 months were older age (odds ratio [OR] = 1.4, 95% confidence interval [CI]: 1.1 to 1.8, per 5-year increase), higher temperature (OR = 1.5, 95% CI: 1.1 to 2.2, per 0.5 degrees C increase), and greater role limitation due to physical functioning (OR = 1.5, 95% CI: 1.2 to 1.9, per 20-point decrease in Short Form-36 score). At 6 months, baseline predictors for failure to recover included female sex (OR = 3.3, 95% CI: 1.0 to 12), a greater number of life events more than 6 months before the disease began (OR = 1.7, 95% CI: 1.1 to 2.5, per each additional life event), and greater family support (OR = 1.9, 95% CI: 1.1 to 4.2, per 7-point increase in social support score). CONCLUSIONS: We were not able to identify objective measures that characterized self-reported failure to recover from acute infectious mononucleosis. The baseline factors associated with self-reported failure to recover at 2 months differed from those associated with failure to recover at 6 months. Future studies should assess the generalizability of these findings and determine whether interventions can hasten recovery. PMID- 11063954 TI - A prospective study of cigarette smoking and the incidence of diabetes mellitus among US male physicians. AB - PURPOSE: To determine the association between cigarette smoking and the incidence of type 2 diabetes mellitus. SUBJECTS AND METHODS: We studied 21,068 US male physicians aged 40 to 84 years in the Physicians' Health Study who were initially free of diagnosed diabetes mellitus, cardiovascular disease, and cancer. Information about cigarette smoking and other risk indicators was obtained at baseline. The primary outcome was reported diagnosis of type 2 diabetes mellitus. RESULTS: During 255,830 person-years of follow-up, 770 new cases of type 2 diabetes mellitus were identified. Smokers had a dose-dependent increased risk of developing type 2 diabetes mellitus: compared with never smokers, the age adjusted relative risk was 2.1 (95% confidence interval [CI]: 1.7 to 2.6) for current smokers of > or = 20 cigarettes per day, 1.4 (95% CI: 1.0 to 2.0) for current smokers of <20 cigarettes per day, and 1.2 (95% CI: 1.0 to 1.4) for past smokers. After multivariate adjustment for body mass index, physical activity, and other risk factors, the relative risks were 1.7 (95% CI: 1.3 to 2.3) for current smokers of > or = 20 cigarettes per day, 1.5 (95% CI: 1.0 to 2.2) for current smokers of <20 cigarettes per day, and 1.1 (95% CI: 1.0 to 1.4) for past smokers. Total pack-years of cigarette smoking was also associated with the risk of type 2 diabetes mellitus (P for trend <0.001). CONCLUSIONS: These prospective data support the hypothesis that cigarette smoking is an independent and modifiable determinant of type 2 diabetes mellitus. PMID- 11063955 TI - Prognostic significance of diabetes as a predictor of survival after cardiac catheterization. APPROACH Investigators. Alberta Provincial Program for Outcome Assessment in Coronary Heart Disease. AB - PURPOSE: Diabetes is a recognized risk factor for the development of cardiac disease, but its importance as a prognostic factor among patients with known cardiovascular disease is less clear. We evaluated survival in patients with and without diabetes who underwent cardiac catheterization for presumed coronary artery disease. SUBJECTS AND METHODS: We analyzed data from a prospective cohort study that captures detailed clinical information and longitudinal outcomes for all patients who undergo cardiac catheterization in Alberta, Canada. We studied 11,468 patients, 1959 (17%) of whom had diabetes. Logistic regression was used to model predictors of 1-year mortality, and proportional hazards analysis was used to model predictors of survival up to 3 years after cardiac catheterization. RESULTS: One-year mortality was 7.6% for patients with diabetes versus 4.1% for those without diabetes (odds ratio = 1.9, 95% confidence interval [CI]: 1.6 to 2.3). After adjusting for other characteristics of the patients, including comorbid conditions, previous cardiac history, coronary anatomy, and renal function, the odds ratio for 1-year mortality was 1.1 (95% CI: 0.8 to 1.3). Similarly, the adjusted hazard ratio for longer term mortality was 1. 2 (95% CI: 1.0 to 1.4, mean follow-up of 702 days). CONCLUSIONS: These results suggest that there is little or no independent association between diabetes and mortality for up to 3 years after cardiac catheterization. Estimates of short- to intermediate term prognosis for diabetic patients with coronary artery disease should be based on the presence of other prognostic factors associated with diabetes. PMID- 11063956 TI - Dietary treatment of hypercholesterolemia: do dietitians do it better? A randomized, controlled trial. AB - PURPOSE: Current guidelines of the National Cholesterol Education Program (NCEP) recommend initial dietary counseling by physicians for most patients with hypercholesterolemia; referral to a registered dietitian and lipid-lowering drugs are recommended only for patients who remain hypercholesterolemic. We evaluated the incremental value of detailed nutritional counseling by dietitians when added to general nutritional advice provided by physicians. SUBJECTS AND METHODS: Hypercholesterolemic patients detected during a cholesterol screening project were randomly assigned to receive dietary counseling by a physician only (70 patients) or by a physician and a registered dietitian (66 patients). Patients were observed for 1 year to determine compliance with NCEP guidelines. RESULTS: At 3 months, the mean (+/- SD) decrease in the serum low-density lipoprotein (LDL) cholesterol level was 7% +/- 11% in the physician group and 12% +/- 10% in the dietitian group (P <0.004). A decrease of 10% or more in the LDL cholesterol level was seen in 25 patients (36%) in the physician group and 43 patients (65%) in the dietitian group (P <0.001). Only 40 (29%) of the patients in both groups achieved their NCEP target goals at 3 months. The majority of these were low-risk patients with an LDL cholesterol target goal of 160 mg/dL. At 12 months, both groups lost about half of the beneficial effects on LDL cholesterol levels, and the difference between the two groups diminished. CONCLUSIONS: The short-term reduction in LDL cholesterol level achieved after counseling by dietitians is superior to that achieved by physicians. However, long-term compliance remains inadequate. For patients at high risk, consideration should be given to a more aggressive dietary approach and possibly earlier introduction of lipid-lowering medications. PMID- 11063957 TI - Abnormalities of coagulation in hypertensive patients with reduced creatinine clearance. AB - PURPOSE: The prothrombotic state that occurs in uremic patients may increase their cardiovascular risk. We studied hypertensive patients with mild-to-moderate impairment of renal function to determine if they had evidence of abnormalities in the coagulation system. SUBJECTS AND METHODS: Renal function was assessed in 382 patients with essential hypertension, in whom 24-hour creatinine clearance, urinary protein excretion, and microalbuminuria were measured. We evaluated the function of the coagulation system by measurement of platelet counts, prothrombin time, partial thromboplastin time, and plasma antithrombin III, fibrinogen, D dimer, and prothrombin fragment 1 + 2 levels. RESULTS: Impaired renal function, defined as a creatinine clearance of 30 to 89 mL per minute per 1.73 m(2) of body surface area, was found in 168 (44%) of the patients. Age, blood pressure, duration of hypertension, and plasma levels of fibrinogen, D-dimer, prothrombin fragment 1 + 2, and lipoprotein(a) were significantly greater in these patients than in those with normal renal function; these differences persisted after adjustment for potential confounders. Creatinine clearance was significantly and inversely correlated with levels of plasma fibrinogen (Spearman's rho = -0.26, P <0.001), D-dimer (rho = -0.33, P <0.001), and prothrombin fragment 1 + 2 (rho = 0.20, P <0.001). Levels of plasma fibrinogen (P = 0.009) and D-dimer (P = 0.003) were correlated with renal function independent of age, blood pressure, duration of hypertension, triglyceride level, urinary protein excretion, and erythrocyte sedimentation rate. Lipoprotein(a) levels were correlated with fibrinogen (rho = 0.16, P = 0.003) and D-dimer (rho = 0.26, P <0.001) levels. CONCLUSIONS: Increased plasma levels of fibrinogen, D-dimer, and prothrombin fragment 1 + 2 are present in hypertensive patients with mildly decreased creatinine clearance, suggesting that the coagulation system is activated in these patients. PMID- 11063958 TI - Effects of nasal continuous positive airway pressure on soluble cell adhesion molecules in patients with obstructive sleep apnea syndrome. AB - PURPOSE: Obstructive sleep apnea syndrome is common in middle-aged men and may be associated with an increased risk of cardiovascular disease. We investigated the effect of nasal continuous positive airway pressure (CPAP) treatment on levels of soluble cell adhesion molecules-which have been shown to be associated with the development of atherosclerosis-in these patients. SUBJECTS AND METHODS: We studied 23 patients with obstructive sleep apnea syndrome diagnosed by polysomnography who were treated with nasal CPAP. Serum soluble intercellular adhesion molecule-1, E-selectin, and vascular cell adhesion molecule-1 levels were measured before nasal CPAP was started, and after 3 or 4 days (n = 19), 1 month (n = 23), or 6 months (n = 11) of treatment. RESULTS: After 3 to 4 days of nasal CPAP therapy, the mean (+/- SD) soluble E-selectin level had decreased from 89 +/- 44 ng/mL to 69 +/- 28 ng/mL (P = 0.002). After 1 month, the soluble intercellular adhesion molecule-1 level had decreased from 311 +/- 116 ng/mL to 249 +/- 74 ng/mL (P = 0.02). After 6 months, soluble vascular cell adhesion molecule-1 levels had not changed significantly, while the mean soluble intercellular adhesion molecule-1 level (212 +/- 59 ng/mL) had decreased further (P = 0.02). Before treatment, soluble intercellular adhesion molecule-1 levels and the apnea and hypopnea index were correlated (r = 0.43, P = 0.04). CONCLUSIONS: Obstructive sleep apnea and hypopnea have a significant adverse effect on serum soluble cell adhesion molecule-1 levels that may be reduced by nasal CPAP treatment. PMID- 11063959 TI - Laboratory testing for infection with the human immunodeficiency virus: established and novel approaches. AB - The enzyme-linked immunosorbent assay (ELISA) and the Western blot are the primary tests for the diagnosis and confirmation of human immunodeficiency virus (HIV) infection. The ELISA, an inexpensive screening test for antibodies to HIV 1, is both sensitive and specific. The HIV-1 Western blot is a reliable confirmatory test following a repeatedly reactive ELISA. False-positive HIV-1 results with this sequence of tests are extremely rare but can occur, and test results that are inconsistent with clinical or other laboratory information should be questioned, repeated, or supplemented. The US Food and Drug Administration has also approved rapid and more accessible testing methods. Oral mucosal transudate and urine testing are noninvasive testing methods; rapid and home sample collection kits offer easier access to testing. PMID- 11063960 TI - Oxidative injury in diseases of the central nervous system: focus on Alzheimer's disease. AB - Alzheimer's disease is one of the most challenging brain disorders and has profound medical and social consequences. It affects approximately 15 million persons worldwide, and many more family members and care givers are touched by the disease. The initiating molecular event(s) is not known, and its pathophysiology is highly complex. However, free radical injury appears to be a fundamental process contributing to the neuronal death seen in the disorder, and this hypothesis is supported by many (although not all) studies using surrogate markers of oxidative damage. In vitro and animal studies suggest that various compounds with antioxidant ability can attenuate the oxidative stress induced by beta-amyloid. Recently, clinical trials have demonstrated potential benefits from treatment with the antioxidants, vitamin E, selegiline, extract of Gingko biloba, and idebenone. Further studies are warranted to confirm these findings and explore the optimum timing and antioxidant combination of such treatments in this therapeutically frustrating disease. PMID- 11063961 TI - Prevalence of upper extremity musculoskeletal disorders in college students. PMID- 11063962 TI - Prevalence of abnormal serum alanine aminotransferase levels in obese patients and patients with type 2 diabetes. PMID- 11063963 TI - Case of the month AB - Each month, we will present a challenging Case of the Month for Green Journal readers, who must use their clinical acumen to arrive at the correct answer. We will also post the case each month on the Journal's web site (http://www.ajmselect.com). Several possible answers may be consistent with the case presentation; use your best judgment. Please send your answer (one per respondent) to The Green Journal at editors@amjmed.org or via Fax to (415) 447 2799. Indicate the case to which you are responding, and include your complete address. The correct answer will appear in the next issue of the Journal. The first five persons who submit correct answers will receive a free one-year subscription to the Journal. Colleagues of Drs. Kashyap in Pune, India are not eligible for this month's case. If you would like to contribute a case, please submit a brief synopsis (<250 words) to the editorial office. PMID- 11063964 TI - New mechanisms for the cardiovascular effects of sleep apnea. PMID- 11063965 TI - Evolving strategies for diagnosing human immunodeficiency virus infection. PMID- 11063966 TI - Doctor, are you there? PMID- 11063967 TI - Structural plasticity: cause, result, or correlate of depression. PMID- 11063968 TI - Regulation of hippocampal neurogenesis in adulthood. AB - A substantial number of new granule neurons are produced in the dentate gyrus in adulthood in a variety of mammalian species, including humans. Numerous studies have demonstrated that the production and survival of new hippocampal neurons can be enhanced or diminished by hormones and experience. Steroid hormones of the ovaries and adrenal glands have been shown to modulate the production of immature neurons by affecting the proliferation of granule cell precursors. Aversive experiences have been demonstrated to decrease the production of immature granule cells, whereas enriching experiences, including learning, have been shown to enhance the survival of new hippocampal cells. These studies indicate that adult generated neurons represent a unique form of structural plasticity that can be regulated by the environment, and furthermore suggest that new neurons play an important role in hippocampal function. PMID- 11063969 TI - Effects of adverse experiences for brain structure and function. AB - Studies of the hippocampus as a target of stress and stress hormones have revealed a considerable degree of structural plasticity in the adult brain. Repeated stress causes shortening and debranching of dendrites in the CA3 region of the hippocampus and suppresses neurogenesis of dentate gyrus granule neurons. Both forms of structural remodeling of the hippocampus appear to be reversible and are mediated by glucocorticoid hormones working in concert with excitatory amino acids (EAA) and N-methyl-D-aspartate (NMDA) receptors, along with transmitters such as serotonin and the GABA-benzodiazepine system. Glucocorticoids, EAA, and NMDA receptors are also involved in neuronal damage and death that is caused in pyramidal neurons by seizures and by ischemia. A similar mechanism may be involved in hippocampal damage caused by severe and prolonged psychosocial stress. Studies using magnetic resonance imaging have shown that there is a selective atrophy of the human hippocampus in a number of psychiatric disorders, as well as during aging in some individuals, accompanied by deficits in declarative, spatial, and contextual memory performance. It is therefore important to appreciate how hippocampal dysfunction may play a role in the symptoms of the psychiatric illness and, from a therapeutic standpoint, to distinguish between a permanent loss of cells and a reversible remodeling to develop treatment strategies to prevent or reverse deficits. Remodeling of the hippocampus may be only the tip of the iceberg; other brain regions may also be affected. PMID- 11063970 TI - Neuronal plasticity and survival in mood disorders. AB - Studies at the basic and clinical levels demonstrate that neuronal atrophy and cell death occur in response to stress and in the brains of depressed patients. Although the mechanisms have yet to be fully elucidated, progress has been made in characterizing the signal transduction cascades that control neuronal atrophy and programmed cell death and that may be involved in the action of antidepressant treatment. These pathways include the cyclic adenosine monophosphate and neurotrophic factor signal transduction cascades. It is notable that these same pathways have been demonstrated to play a pivotal role in cellular models of neural plasticity. This overlap of plasticity and cell survival pathways, together with studies demonstrating that neuronal activity enhances cell survival, suggests that neuronal atrophy and death could result from a disruption of the mechanisms underlying neural plasticity. The role of these pathways and failure of neuronal plasticity in stress-related mood disorders are discussed. PMID- 11063971 TI - Clinical and preclinical evidence for the neurotrophic effects of mood stabilizers: implications for the pathophysiology and treatment of manic depressive illness. AB - Recent neuroimaging studies have demonstrated regional central nervous system volume reductions in mood disorders, findings that are complemented by postmortem observations of cell atrophy and loss. It is thus noteworthy that lithium and valproate have recently been demonstrated to robustly increase the expression of the cytoprotective protein bcl-2 in the central nervous system. Chronic lithium not only exerts neuroprotective effects in several preclinical paradigms but also enhances hippocampal neurogenesis. Valproate robustly promotes neurite outgrowth and activates the ERK mitogen-activated protein kinase pathway, a signaling pathway utilized by many endogenous neurotrophic factors. Consistent with its preclinical neurotrophic/neuroprotective effects, chronic lithium treatment of patients with manic-depressive illness increases brain N-acetylaspartate (a putative marker of neuronal viability and function) levels, an effect that is localized almost exclusively to gray matter. To determine if lithium was producing neuropil increases, quantitative three-dimensional magnetic resonance imaging studies were undertaken, which revealed that chronic lithium significantly increases total gray matter volume in the human brain of patients with manic-depressive illness. Together, these results suggest that a reconceptualization about the optimal long-term treatment of recurrent mood disorders is warranted. Optimal long-term treatment for these severe illnesses may only be achieved by the early use of agents with neurotrophic/neuroprotective effects, irrespective of the primary, symptomatic treatment. PMID- 11063972 TI - The possibility of neurotoxicity in the hippocampus in major depression: a primer on neuron death. AB - A number of studies indicate that prolonged, major depression is associated with a selective loss of hippocampal volume that persists long after the depression has resolved. This review is prompted by two ideas. The first is that overt neuron loss may be a contributing factor to the decrease in hippocampal volume. As such, the first half of this article reviews current knowledge about how hippocampal neurons die during insults, focusing on issues related to the trafficking of glutamate and calcium, glutamate receptor subtypes, oxygen radical generation, programmed cell death, and neuronal defenses. This is meant to orient the reader toward the biology that is likely to underlie any such instances of neuron loss in major depression. The second idea is that glucocorticoids, the adrenal steroids secreted during stress, may play a contributing role to any such neuron loss. The subtypes of depression associated with the hippocampal atrophy typically involve significant hypersecretion of glucocorticoids, and the steroid has a variety of adverse effects in the hippocampus, including causing overt neuron loss. The second half of this article reviews the steps in this cascade of hippocampal neuron death that are regulated by glucocorticoids. PMID- 11063973 TI - Postmortem studies in mood disorders indicate altered numbers of neurons and glial cells. AB - The influence of stress and glucocorticoids on neuronal pathology has been demonstrated in animal and clinical studies. It has been proposed that stress induced changes in the hippocampus may be central to the development of depression in genetically vulnerable individuals. New evidence implicates the prefrontal cortex (PFC) in addition to the hippocampus as a site of neuropathology in depression. The PFC may be involved in stress-mediated neurotoxicity because stress alters PFC functions and glucocorticoid receptors, the PFC is directly interconnected with the hippocampus, and metabolic alterations are present in the PFC in depressed patients. Postmortem studies in major depression and bipolar disorder provide the first evidence for specific neuronal and glial histopathology in mood disorders. Three patterns of morphometric cellular changes are noted: cell loss (subgenual PFC), cell atrophy (dorsolateral PFC and orbitofrontal cortex), and increased numbers of cells (hypothalamus, dorsal raphe nucleus). The relevance of cellular changes in mood disorders to stress and prolonged PFC development and a role of neurotrophic/neuroprotective factors are suggested, and a link between cellular changes and the action of therapeutic drugs is discussed. The precise anatomic localization of dysfunctional neurons and glia in mood disorders may reveal cortical targets for novel antidepressants and mood stabilizers. PMID- 11063974 TI - Effects of early adverse experiences on brain structure and function: clinical implications. AB - Child abuse is associated with markedly elevated rates of major depression and other psychiatric disorders in adulthood. This article reviews preclinical studies examining the effects of early stress, factors that modify the impact of these experiences, and neurobiological changes associated with major depression. Preclinical studies demonstrate that early stress can alter the development of the hypothalamic-pituitary-adrenal axis, hypothalamic and extrahypothalamic corticotropin releasing hormone, monoaminergic, and gamma-aminobutyric acid/benzodiazepine systems. Stress has also been shown to promote structural and functional alterations in brain regions similar to those seen in adults with depression. Emerging data suggest, however, that the long-term effects of early stress can be moderated by genetic factors and the quality of the subsequent caregiving environment. These effects also can be prevented or reversed with various pharmacologic interventions. Preclinical studies of early stress can provide valuable insights in understanding the pathophysiology and treatment of major depression. They also can provide an important tool to use to investigate interactions between genes and environments in determining an individual's sensitivity to stress. More research is needed to understand how inherent factors interact with experiences of abuse and other psychosocial factors to confer vulnerability to develop depression. PMID- 11063975 TI - 3D MRI studies of neuroanatomic changes in unipolar major depression: the role of stress and medical comorbidity. AB - Increasing evidence has accumulated for structural brain changes associated with unipolar recurrent major depression. Studies of neuroanatomic structure in early onset recurrent depression have only recently found evidence for depression associated structural change. Studies using high-resolution three-dimensional magnetic resonance imaging (MRI) are now available to examine smaller brain structures with precision. Brain changes associated with early-onset major depression have been reported in the hippocampus, amygdala, caudate nucleus, putamen, and frontal cortex, structures that are extensively interconnected. They comprise a neuroanatomic circuit that has been termed the limbic-cortical striatal-pallidal-thalamic tract. Of these structures, volume loss in the hippocampus is the only consistently observed change to persist past the resolution of the depression. Possible mechanisms for tissue loss include neuronal loss through exposure to repeated episodes of hypercortisolemia; glial cell loss, resulting in increased vulnerability to glutamate neurotoxicity; stress-induced reduction in neurotrophic factors; and stress-induced reduction in neurogenesis. Many depressed patients, particularly those with late-onset depression, have comorbid physical illnesses producing a high rate of hyperintensities in deep white matter and subcortical gray matter and brain damage to key structures involved in the modulation of emotion. Combining MRI studies with functional studies has the potential to localize abnormalities in blood flow, metabolism, and neurotransmitter receptors and provide a better integrated model of depression. PMID- 11063976 TI - Imaging serotonergic neurotransmission in depression: hippocampal pathophysiology may mirror global brain alterations. AB - The recent development of [carbonyl-(11)C]WAY-100635 for serotonin (5-HT)(1A) and [(18)F]setoperone and [(18)F]altanserin for 5-HT(2A) positron emission tomography receptor imaging has allowed studies of 5-HT neurotransmission in depressive disorders. The hippocampus is likely to be an important brain structure in the pathophysiology of depression because it may mediate both cognitive deficits and hypercortisolemia found in this disorder. Decreased 5-HT(1A) binding was reported in the medial temporal cortex, which receives dense 5-HT innervation, and also throughout neocortical regions. Because the 5-HT(1A) antagonist pindolol may hasten antidepressant effects of selective serotonin reuptake inhibitor medications, its receptor occupancy has been measured in both presynaptic and postsynaptic sites. The results are controversial but suggest that pindolol has preferential occupancy of somatodendritic autoreceptors in the raphe. The results of 5-HT(2A) receptors are mixed, with one showing a significant decrease in the right orbitoinsular cortex and three not detecting a significant change. The disparate findings in patients with depression almost certainly reflect the heterogeneity of the disorder, and we highlight the utility of the hippocampus as a useful target region not only to compare depressed subjects with healthy subjects but also to correlate findings with cognitive function and activity of the limbic-hypothalamic-pituitary axis system. PMID- 11063977 TI - Neuroimaging studies of mood disorders. AB - Neuroimaging studies of major depression have identified neurophysiologic abnormalities in multiple areas of the orbital and medial prefrontal cortex, the amygdala, and related parts of the striatum and thalamus. Some of these abnormalities appear mood state-dependent and are located in regions where cerebral blood flow increases during normal and other pathologic emotional states. These neurophysiologic differences between depressives and control subjects may thus implicate areas where physiologic activity changes to mediate or respond to the emotional, behavioral, and cognitive manifestations of major depressive episodes. Other abnormalities persist following symptom remission, and are found in orbital and medial prefrontal cortex areas where postmortem studies demonstrate reductions in cortex volume and histopathologic changes in primary mood disorders. These areas appear to modulate emotional behavior and stress responses, based upon evidence from brain mapping, lesion analysis, and electrophysiologic studies of humans and/or experimental animals. Dysfunction involving these regions is thus hypothesized to play a role in the pathogenesis of depressive symptoms. Taken together, these findings implicate interconnected neural circuits in which pathologic patterns of neurotransmission may result in the emotional, motivational, cognitive, and behavioral manifestations of primary and secondary affective disorders. PMID- 11063978 TI - Regional metabolic effects of fluoxetine in major depression: serial changes and relationship to clinical response. AB - BACKGROUND: Treatment of major depression with antidepressants is generally associated with a delay in onset of clinical response. Functional brain correlates of this phenomenon have not been previously characterized. METHODS: Time course of changes in brain glucose metabolism were measured using positron emission tomography in hospitalized unipolar depressed patients treated with fluoxetine. Time-specific and response-specific effects were examined at 1 and 6 weeks of treatment. RESULTS: Changes were seen over time, and characterized by three distinct patterns: 1) common changes at 1 and 6 weeks, 2) reversal of the 1 week pattern at 6 weeks, and 3) unique changes seen only after chronic treatment. Fluoxetine responders and nonresponders, similar at 1 week, were differentiated by their 6-week pattern. Clinical improvement was uniquely associated with limbic and striatal decreases (subgenual cingulate, hippocampus, insula, and pallidum) and brain stem and dorsal cortical increases (prefrontal, parietal, anterior, and posterior cingulate). Failed response was associated with a persistent 1-week pattern and absence of either subgenual cingulate or prefrontal changes. CONCLUSIONS: Chronic treatment and clinical response to fluoxetine was associated with a reciprocal pattern of subcortical and limbic decreases and cortical increases. Reversal in the week-1 pattern at 6 weeks suggests a process of adaptation in specific brain regions over time in response to sustained serotonin reuptake inhibition. The inverse patterns in responders and nonresponders also suggests that failure to induce these adaptive changes may underlie treatment nonresponse. PMID- 11063979 TI - Pindolol augmentation of antidepressant treatment: recent contributions from brain imaging studies. AB - Preclinical studies suggest that augmentation of selective serotonin (5-HT) reuptake inhibitors by the 5-HT(1A) receptor agent pindolol might reduce the delay between initiation of treatment and antidepressant response, an effect largely mediated by blockade of 5-HT(1A) autoreceptors in the dorsal raphe nuclei. Although some controlled clinical trials suggest that pindolol might reduce latency to selective serotonin reuptake inhibitor response in acute depressive episodes, the effect is moderate and highly variable. Recent positron emission tomography studies investigating the occupancy of 5-HT(1A) receptors in humans by pindolol have shown that at the dose used most often in clinical trials the occupancy is low and variable, which might explain the inconsistent clinical results. Positron emission tomography studies also suggest that pindolol might be more potent at blocking 5-HT(1A) autoreceptors than at blocking postsynaptic receptors, a property that may be useful in this pharmacologic strategy. Thus, the positron emission tomography data support the potential of pindolol to augment the antidepressant response of selective serotonin reuptake inhibitors, but also imply that this potential has not been fully evaluated. Here we review the clinical trials, the positron emission tomography studies, and the possible mechanisms of pindolol augmentation. It is also suggested that positron emission tomography may be used to define therapeutic dosing early on in the process of clinical evaluation of new treatment strategies. PMID- 11063980 TI - Effects of estradiol and progesterone administration on human serotonin 2A receptor binding: a PET study. AB - BACKGROUND: Preclinical studies demonstrate that 17beta-estradiol (E(2)) increases serotonin-2A receptor (5-HT(2A)R) density in rat frontal cortex. METHODS: We investigated the impact of hormone replacement therapy on 5-HT(2A)R binding potential (BP) using positron emission tomography and [(18)F]altanserin in five postmenopausal women. Subjects were imaged at baseline, following 8 to 14 weeks of transdermal E(2), 0.1 mg/d, and following 2 to 6 weeks of E(2) plus micronized progesterone (P) 100 mg per os twice daily. Regional BPs in the anterior cingulate cortex, dorsolateral prefrontal cortex, and lateral orbitofrontal cortex were calculated by Logan analysis. RESULTS: There was a main effect of time (p = .017) for 5-HT(2A)R BP, which increased 21.2%+/-2.6% following combined E(2) and P administration relative to baseline. This effect was evident in all cerebral cortex regions examined. CONCLUSIONS: 5-HT(2A)R BP increased in widespread areas of the cerebral cortex following combined E(2) + P administration. PMID- 11063981 TI - Glial fibrillary acidic protein immunoreactivity in the prefrontal cortex distinguishes younger from older adults in major depressive disorder. AB - BACKGROUND: Recent postmortem studies in major depressive disorder (MDD) provide evidence for a reduction in the packing density and number of glial cells in different regions of the prefrontal cortex; however, the specific types of glia involved in those morphologic changes are unknown. METHODS: The territory occupied by the astroglial marker glial fibrillary acidic protein (GFAP) was measured as an areal fraction in cortical layers III, IV, and V in sections from the dorsolateral prefrontal cortex (dlPFC) of MDD and control subjects. In addition, the packing density of GFAP-immunoreactive somata was measured by a direct three-dimensional cell counting method. RESULTS: The mean areal fraction and packing density of GFAP-immunoreactive astrocytes in the dlPFC of MDD subjects were not significantly different from those in control subjects; however, in MDD there was a significant strong positive correlation between age and GFAP immunoreactivity. When the MDD group was divided into younger (30-45 years old) and older (46-86) adults, in the five younger MDD adults, areal fraction and packing density were smaller than the smallest values of the control subjects. In contrast, among older MDD subjects these parameters tended to be greater than in the older control subjects. CONCLUSIONS: The present results suggest that the GFAP-immunoreactive astroglia is differentially involved in the pathology of MDD in younger compared with older adults. PMID- 11063982 TI - Neuroglial interactions in a model of para-chlorophenylalanine-induced serotonin depletion. AB - Serotonin (5HT) is involved in the development and plasticity of the CNS through the release of S-100beta, a glial trophic factor which stabilizes synapses and neuronal cytoskeleton and promotes neuronal development. S-100beta is released from glial cells after activation of glial 5HT(1A) receptors. We present in this paper the effects upon neurons and glia of a 5HT depletion induced by 14 days of treatment with para-chlorophenylalanine (PCPA) in adult rats. S-100beta, 5HT, 5HT transporter (5HT-T) and neurofilaments (Nf-200 and Nf-68) expressions were studied by immunohistochemistry and image analysis in striatum, hippocampus, parietal and frontal cortex. Immediately after ending PCPA treatment we found increased intracellular S-100beta immunoreactivity in glial cells, reduced 5HT immunolabelling, reduced density of 5HT-T, Nf-200 and Nf-68 fibers and morphological alterations in neuronal cytoskeleton. One week after PCPA treatment S-100beta immunoreactivity decreased towards control levels, 5HT was normalized in dorsal raphe nucleus, but not in innervation areas; 5HT-T, Nf-200 and Nf-68 fiber densities increased but some neuronal cytoskeletal alterations were still present in striatum. Two weeks after PCPA treatment S-100beta had returned to control levels in most studied regions; 5HT immunoreactivity was normalized, meanwhile 5HT-T, Nf-200 and Nf-68 fiber densities increased reaching values over the control level. We propose that S-100beta could be accumulated in glial cells during the 5HT depletion period, to be released once 5HT levels have recovered. Neuronal cytoskeletal alterations and reduced fiber density may be the expression of decreased extracellular availability of S-100beta. Conversely, increased 5HT T, Nf-200 and Nf-68 expressions, once S-100beta is normalized, may be the biological response to the growth factor release. PMID- 11063983 TI - Decreased expression of nitric oxide synthase in the colonic myenteric plexus of aged rats. AB - Nitric oxide (NO) is a major non-adrenergic, non-cholinergic (NANC) inhibitory neurotransmitter in the gastrointestinal tract. NO released from the myenteric plexus enhances colonic transit and facilitates propulsion of the colonic contents by mediating descending relaxation. Although it has been suggested that colonic transit delays with aging, the mechanism of delayed colonic transit in aging remains unclear. We hypothesized that advanced age is associated with decreased expression of neuronal NO synthase (nNOS) and concomitant reduction in synthesis of NO in the rat colon. We studied nNOS mRNA expression, nNOS immunohistochemistry, nNOS-immunoblotting and NOS catalytic activity in the mid colon obtained from young (age 4-8 months) and aged (age 22-28 months) Fisher (F344xBN)F1 rats. Western blot analysis of PGP 9.5, a generic neuronal marker, of the colonic tissues were employed to study whether the total number of neurons of the myenteric plexus is reduced with aging. The number of nNOS-immunoreactive cells and nNOS synthesis in the colonic myenteric plexus were significantly reduced in aged rats. In contrast, expression of PGP 9.5 in colonic tissues was not affected in aged rats. Northern blot analysis demonstrated that the expression of neuronal nNOS mRNA was significantly reduced in the colonic tissues in aged rats. Basal and veratridine-induced release of L-[(3)H]citrulline were significantly decreased in colonic tissues from aged rats, compared to young rats. It is suggested that advanced age is associated with diminished gene expression of nNOS, nNOS synthesis and catalytic activity of NOS. This may explain the mechanism of delayed colonic transit observed in advanced age. PMID- 11063984 TI - Identification of deflation-sensitive inspiratory neurons in the dorsal respiratory group of the rat. AB - It has been well established that inspiratory neurons of the dorsal respiratory group (DRG) are classified into two types based on whether they receive inputs from slowly-adapting pulmonary stretch receptors (SARs) or not. Inspiratory neurons with SAR inputs are called Ibeta and the others are called Ialpha neurons. In this study, we identified a novel group of inspiratory neurons in Nembutal-anesthetized, paralyzed, and artificially-ventilated rats. (1) These DRG inspiratory neurons were activated characteristically by lung deflation. (2) They were orthodromically activated by electrical stimulation of the vagus nerve at a low intensity just above the threshold for afferents from SARs or rapidly adapting pulmonary stretch receptors (RARs). (3) The orthodromic latencies (ranged from 1.9 to 2.5 ms) indicated that they receive direct inputs from low threshold vagal afferents. (4) Unlike Ibeta neurons, they hardly responded to lung inflation and never exhibited tonic firing in response to maintained lung inflation. (5) The majority (92%) of them were antidromically activated by electrical stimulation of the cervical spinal cord. These deflation-sensitive inspiratory neurons clearly form a distinct group, and their firing pattern is consistent with the hypothesis that they receive inputs from RAR afferents as well as the central inspiratory drive. The results indicating that DRG inspiratory neurons are classified into at least three groups provide new insights into the organization and role of the DRG. PMID- 11063985 TI - Effect of S-adenosyl-L-methionine on rat brain oxidative stress damage in a combined model of permanent focal ischemia and global ischemia-reperfusion. AB - We analyzed the effects of S-adenosyl-L-methionine (SAM) on tissue oxidative status in a combined model of permanent focal ischemia and global reperfusion in the rat brain. The production of thiobarbituric acid reactive substances (TBARS) was measured under basal conditions and after induction with ferrous salt as an indicator of brain lipid peroxidation. Total, oxidized and reduced glutathione were measured as indicators of the antioxidant defense capacity of brain tissue. Mitochondrial reduction of tetraphenyl tetrazolium (TPT) was quantified morphometrically. Results obtained in vitro showed that incubation with SAM reduced lipid peroxidation, with a maximum inhibition of 65.12+/-5.99% after incubation with 1 mmol/l; glutathione production was not significantly modified. In the brain ischemia-reperfusion model, TBARS production increased and glutathione content decreased, and mitochondrial reduction of TPT decreased significantly after ischemia-reperfusion in areas dependent on carotid circulation. The administration of 50 mg/kg SAM per day for 3 days led to the inhibition of brain lipid peroxidation and increased total glutathione production. These changes were accompanied by an increase in mitochondrial capacity to reduce TPT. We conclude that SAM reduces oxidative damage in the rat brain in an experimental model of ischemia-reperfusion. PMID- 11063986 TI - Stimulation of 5-HT(1A) receptors reduces apoptosis after transient forebrain ischemia in the rat. AB - It has recently been shown that 5-HT(1A) receptor stimulation reduced the infarct volume after occlusion of the middle cerebral artery in rats. Since there is increasing evidence that apoptosis is involved in neurodegenerative diseases and stroke, we investigated whether the 5-HT(1A) agonist Bay x 3702 could protect neurons against apoptotic damage in a rat model of transient forebrain cerebral ischemia. Bay x 3702 (4 microg/kg i.v.) caused a 10% reduction of neuronal damage in the hippocampal CA1 subfield. Higher doses of Bay x 3702 (40 and 12 microg/kg i.v.) did not cause any neuroprotective effect, most likely because of the strong reduction of mean arterial blood pressure during the period of Bay x 3702 infusion. Bay x 3702 (4 microg/kg i.v.) diminished DNA laddering in the hippocampus and striatum 4 days after 10 min forebrain ischemia. These results were confirmed by TUNEL-staining. The anti-apoptotic effect was abolished by additional treatment with the 5-HT(1A) receptor antagonist WAY 100635 (1 mg/kg). Taken together, the results suggest that Bay x 3702 can rescue hippocampal as well as striatal neurons from apoptotic cell death in vivo via stimulation of 5 HT(1A) receptors. PMID- 11063987 TI - Hypoxic modulation of striatal lesions induced by administration of endothelin-1. AB - Levels of endothelin-1 (ET-1), a potent endogenous vasoconstrictor, are elevated in plasma and cerebrospinal fluid (CSF) following cerebral ischemia and reperfusion injury. The present study sought insight into the potential differential vasoactive effects on the cerebral vasculature and resultant neural damage of ET-1 during normoxic vs. ischemic conditions and upon reperfusion. Under normoxic conditions, intrastriatal stereotaxic injection of exogenous ET-1 (40 pmol) induced a significant (P<0.05) reduction ( or =0.69, t> or =19, and P<0.001). CONCLUSIONS: Klimek's method of fetal maturity assessment offers a possibility of objective evaluation of maturity immediately after birth. The clinical methods used to date do not have such advantages. Moreover, they require the evaluation of 12-34 parameters, their point range is more complex and their results are given on the scale of 10-50 points. In each of the applied divisions of observed newborns, there was found a high, statistically significant correlation between the indexes evaluating maturity by means of both comparable methods (i.e. Ballard and Klimek scores), which do not take into account weight and fetal age, but refer directly to fetal maturity. The new scale, which has been proposed, is simple and produces comparable results encompassing full maturity in the range of only 6-12 points. PMID- 11064006 TI - Serum ferritin level as a marker of preterm labor. AB - OBJECTIVE: To compare the serum ferritin levels in women with preterm labor (PTL) or preterm premature rupture of membranes with those in normal gravid women. METHOD: The study group consisted of 50 consecutive subjects with preterm labor and 49 subjects with preterm premature rupture of membranes (PROM). The control group consisted of 50 subjects matched with the study group for hemoglobin (Hb) and gestation who did not have PTL or preterm PROM. Serum ferritin levels were assayed in both the groups. RESULTS: Mean serum ferritin levels in patients with preterm labor and preterm premature rupture of membranes were 23.24+/-12.13 ng/ml and 29.44+/-28.41 ng/ml, respectively. The mean serum ferritin in control subjects was 8.69+/-3.7 ng/ml. The difference was evaluated by Student's t-test and was found to be statistically significant. CONCLUSION: The serum ferritin level is significantly raised in pregnant women with preterm labor and preterm PROM. PMID- 11064007 TI - Nucleated red blood cells as a marker in acute and chronic fetal asphyxia. AB - OBJECTIVE: To investigate the variations of nucleated red blood cell (NRBC) counts in acute and chronic fetal hypoxia and to examine if it could be possible to establish a cutoff value for the number of NRBCs for prediction of fetal acidosis. METHOD: We prospectively studied 77 pregnant women. Patients were grouped as acute (n=11) and chronic fetal distress (n=21) and controls (n=45). At delivery the umbilical cord was double clamped and blood samples were collected. RESULTS: The mean NRBC counts in chronic fetal distress group was higher than acute fetal distress. NRBC counts were found to be correlated with umbilical cord pH (r=-0.57; P<0.001). The cutoff value predicting fetal acidosis was determined as 14/100 leukocytes (sensitivity 87%, specificity 81%) by using ROC analysis. CONCLUSION: The duration and the severity of fetal asphyxia may be predicted by the number of NRBCs per leukocyte. PMID- 11064008 TI - Middle cerebral artery to umbilical artery resistance index ratio in the prediction of neonatal outcome. AB - OBJECTIVE: The objectives of this study were to evaluate the usefulness of the middle cerebral artery to umbilical artery resistance index ratio (C/U ratio) as a predictor of adverse perinatal outcome, and to show that the absence of fetal umbilical artery end-diastolic velocity (AEDV) in SGA fetuses is associated with high morbidity and mortality. METHOD: In this prospective study, color Doppler flow imaging was used for the estimation of the C/U ratio in fetuses that were small for their gestational age, in 70 singleton pregnancies between 29 and 42 weeks of gestation. The subjects were categorized into two groups, with Group A consisting of 35 small for gestational age (SGA) fetuses with a normal C/U ratio (1.05 or higher), and Group B comprising 35 SGA fetuses with an abnormal C/U ratio (below 1.05). RESULT: The mean C/U ratio values for birth weight and gestational age were higher in group A than in group B. Fetuses born to mothers in group B stayed longer in the neonatal special care unit (NSCU), whereas the period from ultrasound examination to delivery was higher in the cases in group A. A higher percentage of mothers with an abnormal C/U ratio underwent cesarean section. Fetuses with an absent end-diastolic velocity of the umbilical artery had a higher morbidity. Three stillbirths occurred in fetuses with an absent end diastolic velocity of the umbilical artery. CONCLUSION: Our results suggest that the C/U ratio is a good predictor of neonatal outcome, and could be used to identify fetuses at risk of morbidity and mortality. Fetal umbilical artery AEDV with intrauterine growth restriction is associated with high perinatal morbidity and mortality. PMID- 11064009 TI - Maternal mortality in a tertiary center after introduction of free antenatal care. AB - OBJECTIVE: Determination of maternal mortality rate (MMR) and the main causes of maternal death after the implementation of free antenatal care in a tertiary center in South Africa. METHODS: Retrospective case study on maternal deaths from 1 January 1993 to 31 July 1997. RESULTS: The maternal mortality rate was 128/100000 births. Hypertension disorders (18%), hemorrhage (18%) and sepsis (13%) were the most important causes of death; 44% of all deaths were considered preventable. CONCLUSIONS: The high percentage (44%) of preventable deaths is a cause of concern and is the result of increased workload, decreased staff numbers and late referrals with low socio-economic class of the patient. The discrepancy in the mortality rate between patients booked at the tertiary institution (29.8/100000) and patients booked elsewhere (304.7/100000) or not booked at all (348.5/100000) indicates the need for improving antenatal care in the periphery. PMID- 11064010 TI - Treatment of cervical carcinoma in situ in HIV positive women. AB - OBJECTIVES: To evaluate the safety and effectiveness of hysterectomy vs. cone biopsy in HIV seropositive patients with carcinoma in situ of the cervix (CIS). METHODS: We performed a retrospective case-control study of all HIV seropositive patients diagnosed with carcinoma in situ of the cervix from 1989 to 1995. A control group of HIV(-) women with CIS was also ascertained matched for date of diagnosis of CIS, race and age. RESULTS: There were 439 patients with CIS, of which 45 were HIV seropositive (10.3%). Nine were treated by hysterectomy, 30 by cone biopsy, and six remained untreated. Overall, 63% of HIV(+) patients did not receive any follow-up Pap smear (44% of hysterectomy patients, 50% of cone biopsy patients, and 83% of untreated patients; chi(2) P=0.41). According to Pap smear results, 67% (10/15) cone biopsy patients and 60% (3/5) hysterectomy patients had an abnormal Pap smear after treatment (P=0.9). Median time to recurrence was 12 months in hysterectomy patients vs. 14 months in cone biopsy patients. Deaths occurred in 22% of hysterectomy patients, 17% of cone biopsy patients, and 50% of untreated patients, none due to cervical cancer. Median time to death from presentation was 27.5 months for hysterectomy patients, 11 months for cone biopsy patients, and 7 months for untreated patients (P<0.05). There were no complications in the hysterectomy group, however, two patients were readmitted after cone biopsy for bleeding. When compared to HIV(-) women with CIS, HIV(+) patients were more likely to be treated by hysterectomy (chi(2) P=0.01). CONCLUSION: All patients diagnosed with CIS should be counseled regarding HIV prevention and testing because of a significant seropositive rate. Compliance with gynecologic follow-up is very poor in this patient population. Special efforts should be made to enhance compliance. Cone biopsy and hysterectomy appear to be equally safe and effective in the treatment of CIS. CIS in HIV patients is a poor prognostic indicator for death from any cause. PMID- 11064011 TI - Hypogonadotropic hypogonadism: retrospective analysis of 19 cases. AB - OBJECTIVES: To survey the clinical data of patients with isolated gonadotropin deficiency. METHODS: We retrospectively surveyed the medical records of 19 patients with isolated gonadotropin deficiency aged 16-31 years (mean: 20 years). The major complaint was primary amenorrhea in 100% of the patients, with 42.1% of them also reporting absence of secondary sex traits, and 10% reporting anosmia or hyposmia. Seventy-four percent of the patients had been submitted to hormonal replacement therapy. RESULTS: Bone densitometry was determined in 5 patients and revealed lumbar spine osteopenia in 3 patients and femoral osteopenia in 1. An association with urologic malformations was detected in 10.5% of cases and an association with gynecologic malformations was detected in 31.6%. CONCLUSIONS: Isolated gonadotropin deficiency can be easily diagnosed but requires early estrogen replacement therapy because of a higher risk of osteopenia and consequently of osteoporosis. Concomitant urogenital malformations are frequent and should be investigated. PMID- 11064012 TI - Low pulse oxygen saturation in post-menopausal women at high altitude is related to a high serum testosterone/estradiol ratio. AB - OBJECTIVE: The present study was designed to determine if low pulse oxygen saturation (SpO(2)) in post-menopausal women at high altitude was related to high serum testosterone/estradiol (T/E(2)) ratio. METHOD: Studies were carried out in 191 women living in Cerro de Pasco, Peru (4340 m above sea level) and in 56 women living in Lima (150 m asl). Body weight, height, SpO(2), hematocrit, serum follicle stimulating hormone (FSH), T and E(2) levels were measured in all women. Body mass index (BMI), menopausal status and serum T/E(2) ratio were calculated from data. RESULT: At high altitude, the values of SpO(2), serum T, and serum E(2) were significantly lower in post-menopausal than in pre-menopausal women. Hematocrit, serum FSH levels, and serum T/E(2) ratios were significantly higher in post-menopausal women. Serum T and E(2) levels, T/E(2) ratio, SpO(2), and hematocrit levels were not further changed with time after menopause. Women with SpO(2)<85% had lower serum E(2), a higher serum T/E(2) ratio, and higher hematocrit values than women with SpO(2)>90%. At high altitude, multiple regression analysis showed that low SpO(2) was related to menopausal status ( 2.6+/-0.83; beta+/-S.E.; P<0.002) and not to chronological age (-0.06+/-0.04; P: NS). Further analysis showed that low SpO(2) was related to high basal serum FSH levels and a high serum T/E(2) ratio in the presence of an interaction between FSH and T/E(2). CONCLUSION: Low values of SpO(2) in women at high altitude were related to a high T/E(2) ratio. PMID- 11064013 TI - Arteriovenous malformations of the uterus associated with medical abortion. AB - Seven cases of uterine arteriovenous malformations associated with pregnancy were initially suspected by the history of prolonged bleeding after a medical abortion and then confirmed by color Doppler scanning. All cases were managed expectantly and resolved spontaneously. When AVMs required diagnosis by pathology or angiography, only the most severe cases would have been reported. Now that they can be recognized on ultrasound, it is possible that mild cases such as these will be found to be much more common. PMID- 11064014 TI - Decision-directed hysterectomy: a possible approach to improve medical and economic outcomes. AB - OBJECTIVE: This study was undertaken to determine if the use of formal guidelines in selecting the route of hysterectomy would improve medical and economic outcomes. METHOD: Data from 4595 hysterectomies performed at a single center in women whose primary diagnosis were unrelated to invasive cancer or pregnancy were analyzed in terms of mean, uterine weight, costs, length of stay, and complications. RESULTS: When formal guidelines were used to determine the route of hysterectomy, vaginal hysterectomy was performed in 90% of the patients treated and in 100% of the patients in whom the pathology was confined to the uterus. In comparison, when formal guidelines were not incorporated in the decision-making process, vaginal hysterectomy was performed in 42% of the patients treated and in 64% of the patients in whom the pathology was confined to the uterus. CONCLUSIONS: Using these or similar guidelines to assist in clinical decision making would have resulted in a potential savings of US$1184000 for every 1000 hysterectomies performed at the institution where this study was undertaken and would have freed up 1020 patient-bed days and reduced complications by approximately 20%. PMID- 11064015 TI - Two years prospective study of perinatal mortality in Jos, Nigeria. PMID- 11064016 TI - Treatment with antithrombin III (AT III) concentrate in pre-eclampsia. PMID- 11064017 TI - Primary ovarian pregnancy. PMID- 11064018 TI - Anatomical features of the uterosacral ligaments in patients with congenital pelvic anomalies. PMID- 11064019 TI - The role of glomeruli in the neural representation of odours: results from optical recording studies. AB - Odours are received by olfactory receptors, which send their axons to the first sensory neuropils, the antennal lobes (in insects) or the olfactory bulb (in vertebrates). From here, processed olfactory information is relayed to higher order brain centres. A striking similarity in olfactory systems across animal phyla is the presence of glomeruli in this first sensory neuropil. Various experiments have shown that odours elicit a mosaic of activated glomeruli, suggesting that odour quality is coded in an 'across-glomeruli' activity code. In recent years, studies using optical recording techniques have greatly improved our understanding of the resulting 'across-glomeruli pattern', making it possible to simultaneously measure responses in several, often identifiable, glomeruli. For the honeybee Apis mellifera, a functional atlas of odour representation is being created: in this atlas, the glomeruli that are activated by different odorants are named. However, several limitations remain to be investigated. In this paper, we review what optical recording of odour-evoked glomerular activity patterns has revealed so far, and discuss the necessary next steps, with emphasis on the honeybee. PMID- 11064020 TI - Protein catabolism in mosquitoes: ureotely and uricotely in larval and imaginal Aedes aegypti. AB - Catabolism of excess dietary protein by Aedes aegypti was investigated during larval development, during and after metamorphosis. Activity profiles were established for xanthine dehydrogenase (XDH, uricotelic pathway) and arginase (ureotelic pathway). Both enzymes are active at all times during the life-cycle. During the aquatic larval and pupal instars, XDH and arginase activities increase with body size. Maximal activities of these two enzyme systems coincide with the time of metamorphic restructuring.Both enzymes are found in the fatbody tissue: XDH activity is found in 80% of the tissue, while arginase activity is distributed equally between abdominal fatbody and the thorax. This might indicate a role for arginase other than catabolic, such as energy metabolism.Arginase activity is high in the aquatic instars and low in sugar-fed females but increases after blood-feeding. XDH activity, also high in larvae and pupae, increases markedly after a blood meal.Larval excretion is characterized by the ureotelic pathway. The pupae as closed systems excrete neither uric acid nor urea; urate accumulates during larval and pupal periods, is conserved throughout metamorphosis, and is finally voided with the meconium by the teneral imago. This presents a form of transient storage-excretion. PMID- 11064021 TI - Apocrine secretion of amylase and exocytosis of trypsin along the midgut of Tenebrio molitor larvae. AB - Amylase and trypsin were purified from Tenebrio molitor midgut larvae and used to raise antibodies in a rabbit. A Western blot of T. molitor midgut homogenates, after sodium dodecyl sulfate-polyacrylamide gel electrophoresis using amylase and trypsin antisera, showed only bands co-migrating with the purified enzymes. The antisera were used to localize the enzymes by immunofluorescence and immunogold labeling. Amylase occurs in a few regularly disposed anterior midgut cells. Non amylase-secreting anterior midgut cells are proposed to be water-absorbing cells based on morphology and dye movements. Amylase is found inside vesicles originating from Golgi areas that seem to fuse together before their release along with the now disorganized apical cytoplasm (apocrine secretion). Trypsin precursors are observed inside small vesicles near the apical plasma membrane of posterior midgut cells, suggesting an exocytic mechanism of secretion, followed by putative trypsin activation. Apocrine secretion is thought to be an adaptation to enhance the dispersion of secretory vesicle contents released from a water absorbing epithelium, whereas exocytosis is an efficient secretory mechanism in a water-secreting epithelium. PMID- 11064022 TI - Relationships between cold hardiness and diapause, and between glycerol and free amino acid contents in overwintering larvae of the oriental corn borer, Ostrinia furnacalis. AB - To elucidate the relationship between diapause and cold hardiness in the oriental corn borer, Ostrinia furnacalis, the levels of various substances, cold hardiness and respiration were measured in diapausing and post-diapausing overwintering larvae. Under field conditions, diapause terminated between November and January, although O(2) consumption, measured at 20 degrees C in the laboratory, remained at a high level from October to January. Glycerol content was low during October and November but greatly increased during December and January. Serine was the most abundant of the free amino acids, and its concentrations were especially high during October and November, while the concentration of alanine increased in December and January. Under laboratory conditions, glycerol levels were low in diapausing larvae, and in post-diapausing larvae that were acclimated at either high temperatures or under anaerobic conditions, while they were high in post diapausing larvae kept under aerobic, low temperature conditions. The survival rate (cold hardiness) was strongly correlated with glycerol content but not with serine or alanine levels. These results suggest that O. furnacalis has a highly developed cold hardiness mechanism in which termination of diapause enables the larvae to increase glycerol levels when the temperature decreases. PMID- 11064023 TI - Haemocyte changes in resistant and susceptible strains of D. melanogaster caused by virulent and avirulent strains of the parasitic wasp Leptopilina boulardi. AB - Two strains of Drosophila melanogaster (resistant and susceptible) were parasitized by a virulent or avirulent strain of the parasitoid wasp Leptopilina boulardi. The success of encapsulation depends on both the genetic status of the host strain and the genetic status of the parasitoid strain: the immune cellular reaction (capsule) is observed only with the resistant strain-avirulent strain combination. The total numbers of host haemocytes increased in all 4 combinations, suggesting that an immune reaction was triggered in all hosts. Resistant host larvae infected with the virulent or avirulent strains of parasitoid wasp had slightly more haemocytes per mm(3) than did susceptible host larvae at the beginning of the reaction (less than 15 h post-parasitization). This difference disappeared later. Only the virulent parasitoid strain caused the production of a high percentage of altered lamellocytes (from a discoid shape to a bipolar shape), half the total number of lamellocytes are altered. This suggests that the alteration of lamellocyte shape alone is not sufficient to explain the lack of capsule formation seen in resistant hosts parasitized by the virulent strain. Lastly, there were very few altered lamellocytes in resistant or susceptible hosts parasitized by the avirulent parasitoid strain, two combinations in which no capsule was formed. As is now established for Drosophila parasitoid interactions, virus-like particles contained in the long gland of the female wasp affect the morphology of the lamellocytes. The results presented here are further proof of the action (direct or indirect) of virus like particles of the virulent strain on lamellocytes. PMID- 11064024 TI - Diapause in the gypsy moth: a model of inhibition and development. AB - We present here the first recorded age-specific estimates of the developmental response to temperature in diapausing gypsy moth, Lymantria dispar (L.). The effect of temperature on diapause development in gypsy moth eggs was examined by exposing individual eggs to temperature regimes of 5 degrees C interrupted by a single, brief exposure to an experimental temperature. Exposure to each of six experimental temperatures took place at six different times during diapause. The relative effect of the exposure on diapause development was estimated by comparing the duration of diapause in each of the treatments to the duration in a control treatment of constant 5 degrees C. The effect of each temperature did not remain constant throughout the diapause phase and the pattern of change was not uniform among the experimental temperatures. We propose a model of diapause where the developmental phase is controlled by two simultaneous temperature-dependent processes: a typical developmental response to temperature that is inhibited by a temperature-activated biochemical agent, and the temperature-dependent removal of the inhibiting agent. This simple model of two simultaneous and temperature dependent processes explains 92% of the variability in diapause duration observed in the experimental regimes. PMID- 11064025 TI - An antidiuretic factor in the forest ant: purification and physiological effects on the Malpighian tubules. AB - Formica polyctena antidiuretic factor (FopADF) was purified from a 15% trifluoroacetic acid (TFA) extract of the abdomens of 150,000 worker ants. After solid phase extraction of the crude extract and reversed-phase HPLC on two C(18) columns, an antidiuretic factor was isolated. Tested at a concentration of 1.0 ant-equivalents/ul (ant-eq/ul), the factor reversibly inhibited fluid secretion of isolated Malpighian tubules to 29+/-5% (mean+/-SE, n=24) of the control value. The same concentration of FopADF reversibly depolarized both the basolateral membrane potential (V(bl)), from -21+/-2 mV to -3+/-1 mV (n=5), and the apical membrane potential (V(ap)), from -65+/-5 mV to -20+/-5 mV (n=5). Similar effects on fluid secretion and V(ap) were caused by a TFA extract of the haemolymph of ants with non-secreting tubules. Unfortunately, further purification of FopADF on a C(4) column led to a loss of activity in the fluid secretion assay. This is the first time an endogenous antidiuretic factor acting directly on Malpighian tubules has been partially purified and shown to depolarize the tubule cell membranes. PMID- 11064026 TI - Effects of age and blood sugar levels on the proboscis extension of the blow fly Phormia regina. AB - In some insects the proboscis is extended to imbibe a sugar solution if the concentration of sugar applied to the chemosensilla exceeds the behavioural threshold value. Recently, I found a reversal of the threshold values of this "proboscis extension reflex" (PER) in the blow fly (Phormia regina M.) for glucose and fructose. It depended on maturation and physiological conditions, both of which are explicable in terms of changing concentration of haemolymph trehalose. The direct injection of trehalose into the fly haemocoele brought about a dramatic shift of the threshold values of PER measured on tarsi or labellar sensilla, suggesting a strong dependence of PER on the blood sugar level. Using the tip-recording method, the dose-response (impulse frequency) curves for glucose and fructose were obtained on individual largest labellar chemosensilla. The curves for glucose and fructose crossed at one point because the former had a steeper gradient and higher maximum response than the latter. Injection experiments with trehalose were also carried out to test for changes in gustatory response. The shifting of the behavioural dose-response curves for glucose and fructose two hours after injection of 1 M trehalose (2 ul) into the haemocoele of the fly was associated with significant reduction in responsiveness of labellar chemosensilla to glucose, but less so to fructose. No change in responsiveness was found following injection of mannose. A hypothesis to explain the reversal relation of the PER thresholds, based on a shift in the firing rate in gustatory sensilla and possibly also interneurons, is discussed. PMID- 11064027 TI - Blue biliprotein as an effective factor for cryptic colouration in Rhodinia fugax larvae. AB - The fifth instar larva of the saturniid silkworm, Rhodinia fugax, is light yellowish-green on its dorsal surface and dark green on the ventral surface with a lateral demarcation between the two colours. The larva of R. fugax closely resembles the leaves of the host plant, Quercus serrata, in colour and shape. The spectral reflectance of the larval integument of R. fugax corresponds to that of the Q. serrata leaf. In the larval integument, there is more blue biliproteins (BPs) on the ventral surface than on the dorsal surface. Light intensity influences larval colouration. The larval integuments are green under light conditions (1000 lux), whereas larvae kept in dark conditions (10 lux) turn yellow. The BP-I content of the haemolymph is also affected by light intensity. The quantities of BP-I and its blue chromophore are higher under light conditions than under dark conditions. In contrast, there is little difference in the yellow chromophore content between the two light intensities. When larvae are kept in the light, the BP-I content in the cuticle is higher than under dark conditions in both the ventral and dorsal surfaces, and its chromophore content parallels the BP content. However, the amounts of BP-II and its chromophore in the epidermis show no change with the light intensity. Moreover, the quantity of yellow chromophore in the integument is also not affected by light intensity. Therefore, light stimulates the accumulation of BP-I and its chromophore in the haemolymph and cuticle, whereas the accumulation of BP-II and its chromophore in the epidermis are not influenced by light intensity. These results suggest that BPs and their chromophores determine the larval colouration and may play an important role in the cryptic colouration of R. fugax larvae. PMID- 11064038 TI - Leaf variations in Elaeagnus angustifolia related to environmental heterogeneity. AB - Elaeagnus angustifolia (Russian olive) is a Eurasian tree that has become naturalized and has invaded zones along watercourses in many arid and semiarid regions of the world. These habitats are characterized by vertical environmental gradients, thus trees must develop some plasticity to adapt to the wide range of site conditions. This study was undertaken to test the hypothesis that variations in leaf anatomy and morphology of E. angustifolia reflect their adaptability to the differences in the microclimate that occur within the canopy of single trees. Foliar architecture, blade and petiole epidermal and internal anatomy were examined in leaves at different canopy positions and related to environmental conditions. Upper sun-leaves are exposed to higher solar irradiance and lower air humidity and are smaller, more slender and thicker than the lower, half-exposed and shade-leaves. Color varies between the leaves at different levels, from silvery grey-green in the upper strata, to dark green in the lower one. Bicolor is more evident in half-exposed and shaded leaves. When compared with the lower half-exposed and shade-leaves, the upper leaves of E. angustifolia have a greater areole density, a higher mesophyll proportion and stomatal density. Trichomes are multicellular, pedestalled, stellate-branched or peltate and their form and density can be associated with leaf color and appearance. The slender petioles of the upper leaves have proportionally more epidermis, collenchyma and phloem and less parenchyma and xylem than those of lower leaves, when observed in transverse sections. Foliar morphological and anatomical variability in E. angustifolia may be considered an adaptive advantage that enables leaves to develop and function in habitats marked by strong variations of solar radiation, air temperature and humidity. PMID- 11064039 TI - The influence of elevated carbon dioxide and water availability on herbaceous weed development and growth of transplanted loblolly pine (Pinus taeda). AB - Loblolly pine (Pinus taeda L.) seedlings were grown in competition with native weeds using soil and seed bank collected from recently chopped and burned areas near Appomattox, Virginia. One-year-old seedlings were planted and weeds allowed to germinate from the native seed bank while being exposed to CO(2) (ambient and elevated - approximately 700 ppm) and water (water stressed and well watered) treatments for approximately one growing season in a greenhouse. Elevated CO(2) did not influence total weed biomass; however, C(3) weed community development was favored over C(4) weed community development in elevated CO(2) regardless of water availability. This suggests that weed community composition may shift toward C(3) plants in a future elevated CO(2) atmosphere. Pine growth was significantly greater in the well watered and elevated CO(2) treatments compared to the water stressed and ambient treatments, respectively, even though they were competing with native herbaceous weeds for resources. There was a significant water and CO(2) interaction for pine root:shoot ratio. Under elevated CO(2), root:shoot ratio was significantly greater in the water stressed treatment than the well watered treatment. In contrast, there was no significant difference in the root:shoot ratio under the ambient CO(2) treatment for either water treatment. These results suggest that loblolly pine seedlings will respond favorably in an elevated CO(2) atmosphere, even under dry conditions and competing with herbaceous weeds. PMID- 11064040 TI - Growth and nitrogen metabolism of Catasetum fimbriatum (orchidaceae) grown with different nitrogen sources. AB - Catasetum fimbriatum is an epiphytic orchid from South America that has been used for 15 years as a model plant for metabolic and developmental studies in our laboratory. In this work, C. fimbriatum plants were aseptically grown with 6 mol m(-3) of either glutamine or inorganic nitrogen forms (NO(3)(-):NH(4)(+) ratios). The highest biomass accumulation was found in plants supplied with glutamine; no significant difference was observed in plants incubated in the presence of inorganic nitrogen sources. Nitrogen assimilation was limited in the presence NO(3)(-) as a sole nitrogen source. C. fimbriatum did not accumulate NO(3)(-) and very low rates of in vivo nitrate reductase activity were observed. Most nitrate reductase activity (70%) was detected in the 2 cm apical roots. Nitrate-treated plants exhibited relatively lower amounts of free amino-N, chlorophyll and free NH(4)(+) contents and higher soluble sugar contents than the NH(4)(+)-treated plants. While shoot glutamine synthetase activity was only slightly affected by nitrogen sources, root glutamine synthetase activity was not modified by any nitrogen form. Glutamate dehydrogenase-NADH activity in shoot tissues was not influenced by any nitrogen source. However, the glutamate dehydrogenase-NADH activity in roots was enhanced when NH(4)(+) tissue contents was augmented by increasing NH(4)(+) in the medium and by the presence of glutamine. Our results strongly suggest that organic nitrogen and NH(4)(+) are probably the most important nitrogen sources to C. fimbriatum plants. PMID- 11064041 TI - Uptake capacity of amino acids by ten grasses and forbs in relation to soil acidity and nitrogen availability. AB - Uptake capacity of organic nitrogen was studied in solution experiments on eight grasses and two forbs growing in acid soils with relatively high nitrogen mineralisation in southern Sweden. Uptake of a mixture of amino acids (alanine, glutamine, glycine), that varied between 1.6 and 6.3 umol g(-1) dw root h(-1), could not be explained by soil data from the species' field distributions (pH, total carbon and nitrogen, potential net mineralisation of ammonium and nitrate). The ratio between organic and inorganic nitrogen (methylamine) uptake was <0.05 for the forbs, higher for the grasses with a maximum of 1.42 for Deschampsia flexuosa. The ratio was negatively correlated with measures related to soil acidity (Ellenberg's R-value, soil nitrate and total carbon) but not, as hypothesised, with the total amount of mineralised nitrogen. The total demand on nitrogen by all components of the ecosystem would probably have described the extent to which competition among and between plants and microbes induced nitrogen limitation. In a methodological study two grasses were exposed to pH 3.8, 4.5 and 6.0 and to 50, 100 and 250 umol l(-1) of three amino acids. Uptake was also compared between intact plants and excised roots. The treatment response varied considerably between the species which stresses the importance of studying intact plants at field-relevant pH and concentrations. PMID- 11064042 TI - Classical and biochemical endpoints in the evaluation of phytotoxic effects caused by the herbicide trichloroacetate. AB - Three terrestrial plant species, oat (Avena sativa ), Chinese cabbage (Brassica campestris cv. chinensis) and lettuce (Lactuca sativa), were exposed to different concentrations of herbicide TCA (sodium trichloroacetate) in a growth test according to guideline OECD # 208. Classical (i.e. germination and biomass) and biochemical (i.e., antioxydant enzyme activities) endpoints were investigated. Germination rate decreased significantly at 3.9 mg TCA kg dry soil(-1) (for oat and lettuce) and 62.5 mg TCA kg dry soil(-1) (for Chinese cabbage). Biomass decreased significantly only at 1.9 mg TCA kg dry soil(-1) (for oat and lettuce) and 15.6 mg TCA kg dry soil(-1) (for Chinese cabbage). The activities of superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7) and glutathione reductase (EC 1.6.4.2) increased significantly at the lowest concentration of TCA tested, i.e. 0.03 mg TCA kg dry soil(-1) (for oat and lettuce) and 0.48 mg TCA kg dry soil(-1) (for Chinese cabbage). Our results showed a ranking of sensitivity among the different endpoints for the three plant species: enzyme activities>biomass>germination rate. The increase in antioxidant enzyme activities observed in this study ensured the detoxification of increased levels of active oxygen species, and presumably prevented the plants from undergoing oxidative stress damage. Thus, the use of enzyme activities will permit the detection of early injury in plant growth testing. PMID- 11064043 TI - The influence of elevated ultraviolet-B radiation (UV-B) on tissue quality and decomposition of loblolly pine (Pinus taeda L.) needles. AB - Stratospheric ozone depletion is expected to elevate the influx of ultraviolet-B radiation (UV-B) to the biosphere. Increased levels of UV-B may, in turn, alter important ecosystem processes such as decomposition. Previous studies have shown that growth under elevated UV-B can alter leaf quality in angiosperm species and thereby indirectly change subsequent rates of leaf decay. In this experiment, we determined if elevated UV-B would alter the chemical composition and decay of needle tissue from two seed sources of the gymnosperm Pinus taeda L. Maryland and Virginia seed sources of P. taeda were grown in the field for 3 years beneath lampbanks supplying either ambient, low elevated or high elevated UV-B. These levels of UV-B corresponded to 0, 16 and 25% stratospheric ozone depletion at the experimental site in Beltsville, MD (39 degrees N). Needles were collected from six randomly chosen plants for each combination of seed source and UV-B level. The needle samples were analyzed for total C and N, UV-B absorbing compounds, and carbon fractions. Decay rates were also determined by measuring rates of CO(2) evolution from needle material decomposed under laboratory conditions. UV-B did not significantly alter the chemical composition of needles from the Virginia seed source. In contrast, needles from the Maryland seed source tended to have elevated lignin/N ratios and a lower holocellulose content when grown under the highest level of UV-B. Furthermore, while needles from the Virginia pines did not have UV-B altered decay rates, Maryland needles grown under low elevated UV-B conditions decomposed 36% more rapidly than needles from other treatments. Results from this experiment illustrate at least three characteristics about the indirect effect of UV-B on decomposition, (1) UV-B can modify decomposition of tissue from gymnosperms as well as angiosperms; (2) UV-B effects on tissue chemistry and decay may not only be species-specific but also seed-source specific; and (3) UV-B effects on decomposition may not increase with increasing UV-B dose. PMID- 11064044 TI - Gas-exchange response and stomatal and non-stomatal limitations to carbon assimilation of sunflower under salinity. AB - Sunflower (Helianthus annuus) was grown in both open-field and outdoor potted conditions in Southern Italy, and irrigated with water having electrical conductivity ranging between 0.9 and 15.6 dS m(-1) obtained by different NaCl concentrations. The aim of the work was to study the leaf area and photosynthetic responses of sunflower to mild salt stress. The response curve (A/c(i)) of assimilation (A) to leaf internal CO(2) concentration (c(i)) was used to determine leaf gas-exchange parameters, in order to evaluate stomatal and non stomatal limitations to photosynthesis in relation to salt stress. In the field, a reduction of 19% in leaf area expansion occurred, while no correlation was observed between Psi(l) and stomatal conductance to water vapour (g(sw)) ranging between 0.76 and 1.35 mol m(-2) s(-1). This result was also evident at a higher salinity level reached in the pot experiment where leaf osmotic potential (psi(s)) varied from -1.35 to -2.67 MPa as compared with the field experiment, where psi(s) ranged from -1.15 to -1.42 MPa. Considering the two experiments as a unique data set, the assimilation rate, the stomatal conductance to CO(2) (g(sc)) and the sensitivity of A to c(i) variation (g*) were not significantly influenced by salinity in the whole range of psi(s). As a consequence, the stomatal and non stomatal limitations to photosynthesis were not affected by salt treatment, averaging around 20 and 80%, respectively. The variation in A (from 44 to 29 umol m(-2) s(-1)) was paralleled by the variation in g(sc) (from 0.47 to 0.84 mol m( 2) s(-1)), with a remarkable constancy of both c(i) (200+/-12.5 umol mol(-1)) and normalized water-use efficiency (5+/-0.7 umol mmol(-1) kPa), showing the optimal behaviour of the plant processes. These findings indicate that, under mild salt stress, the same as observed under water deficit, sunflower controls assimilation mainly by modulating leaf area rather than by stomatal closure, and that non stomatal limitation of photosynthesis was not affected at all by the level of salinity reached in this study. PMID- 11064045 TI - Human cytomegalovirus detection by a quartz crystal microbalance immunosensor. AB - A piezoelectric affinity sensor has been developed to detect distinctive antigens of the human cytomegalovirus. Either the specific antibodies or the antigen were immobilized on the gold electrode. To develop a rapid immunoassay, various assay formats were tested in relation with the different antigen composition. First, a direct assay was carried out immobilizing the specific antibody on the crystal surface by passive adsorption. Next, Protein A, thiol/poly L-lysine mixed self assembled monolayers were tested as methods of gold modification. A competitive format was exploited by immobilization of the antigen onto the crystal activated by SAM and poly L-lysine. This procedure yielded a preliminary calibration curve. A linear range between 2.5 and 5 ug/ml of gB epitope in solution and a detection limit of 1 ug/ml were measured. PMID- 11064046 TI - Synthesis of methyl beta-D-fructoside catalyzed by levansucrase from Rahnella aquatilis. AB - Methyl beta-D-fructoside(MF) was formed from sucrose and methanol by a transfructosylation reaction using recombinant levansucrase from Rahnella aquatilis. The increase in the yield of MF formation was achieved by increasing methanol concentration. The enzyme stability at higher concentrations of methanol was maintained by lowering the reaction temperature. The optimum temperature and sucrose concentration for MF formation was 10 degrees C and 50 gL(-1) respectively and the yield of MF was 70%. PMID- 11064047 TI - Model-based bioreactor selection for large-scale solid-state cultivation of Coniothyrium minitans spores on oats. AB - Non-mixed and mixed SSF reactors were evaluated for their applicability in large scale spore production of the biocontrol fungus Coniothyrium minitans. The major problem to overcome in large-scale SSF is heat accumulation. Testing various cooling strategies in large-scale bioreactors would be very expensive and time consuming, therefore lab experiments in combination with mathematical simulations were used instead. The metabolic heat production rate, estimated from the oxygen consumption rate of C. minitans on oats in Erlenmeyer flasks, was about 500 Watt per m(3) bed. Conductive cooling in packed-bed reactors was insufficient to cool large reactor volumes (radius > 0.2 m). The poor thermal conductivity of the bed (lambda(b) = 0.1 W m(-2) K(-1)) resulted in steep radial temperature profiles. Adequate temperature control could be achieved with forced aeration, but concomitant water losses lead to significant shrinkage of the oats (30%) and critically low water activities, even though the bed was assumed to be aerated with water saturated air. Mixed systems, however, allowed heat removal without the need of evaporative cooling. Simulations showed that large volumes could be cooled via the wall at low mixing intensities and small temperature driving forces. Experimental studies showed no detrimental effect of mixing on spore production by C. minitans. The spore production yield in a continuously mixed scraped-drum reactor (0.2 rpm) was 5 x 10(12) spores per kg dry oats after 450 hours. Based on the scale-up potential of the mixed system and the absence of detrimental mixing effects it is believed that a mixed bioreactor is superior to a non-mixed system for large-scale production of C. minitans spores. PMID- 11064048 TI - Surface hydrophobic amino acid residues in cellulase molecules as a structural factor responsible for their high denim-washing performance. AB - The denim-washing performance of six purified fungal cellulases (four endo-1,4 beta-D-glucanases and two cellobiohydrolases) was compared using a model microassay. The performance of cellobiohydrolases per mg of protein was much lower than that of endoglucanases. For endoglucanases, it varied up to 5 times between the best and the worst enzyme. Experiments with amino acids immobilized on cross-linked agarose showed that their side chains may bind indigo owing to hydrophobic interactions and formation of hydrogen bonds. The best binding effects provided Tyr and Phe. Analysis of three-dimensional structures of cellulase molecules showed that a certain correlation exists between the washing performance of enzyme and (i) quantity (percentage) of aromatic residues exposed to solvent on the surface of protein globule or (ii) overall percentage of the surface hydrophobic residues. Data presented provide an evidence that the molecules of certain cellulases, which have hydrophobic domains (clusters of closely located non-polar residues) on their surface, may bind indigo and thus act as emulsifiers helping the dye to float out of cellulose fibers to the bulk solution. PMID- 11064049 TI - Simultaneous purification and immobilization of Aspergillus niger xylanase on the reversibly soluble polymer Eudragit(TM) L-100. AB - The non-covalent immobilization of a commercial preparation of xylanase from A. niger was carried out on a reversibly soluble-insoluble enteric polymer Eudragit(TM) L-100. The immobilization of the xylanase activity by adsorption was simultaneously accompanied by removal of cellulase activity since the latter did not bind to the polymer. Thus, the soluble enzyme derivative may be useful for treatment of paper pulp bleaching in paper industry. The immobilized xylanase retained 60% of its activity toward xylan as the substrate. No change was observed in the pH optimum (5.5) of the enzyme upon immobilization. Only marginal increase in the K(m) of the free enzyme (3.6 mg ml(-1) to 5.0 mg ml(-1)) upon immobilization on the soluble polymer reflected that the enzyme-substrate binding continues to be efficient in spite of the macromolecular nature of the substrate. Fluorescence spectroscopy and UV difference spectroscopy were used to probe the change(s) in the enzyme structure upon immobilization. This change in structure was correlated with the "effectiveness factor" of the enzyme activity. CD spectra also showed that the enzyme undergoes drastic changes in the structure. PMID- 11064050 TI - Oxygen transfer and uptake rates during xanthan gum production. AB - Oxygen uptake rate and oxygen mass transfer rate have been studied during xanthan gum production process in stirred tank bioreactor. Empirical equations for the oxygen mass transfer coefficient have been obtained taking into account several variables such as air flow rate, stirrer speed and apparent viscosity. Oxygen uptake rate evolution in the course fermentation has been measured, obtaining an equation as a function of biomass concentration, including overall growth and non growth-associated oxygen uptake. A metabolic kinetic model has been employed for xanthan gum production description including oxygen mass transfer and uptake rates. The results point out that this model is able to describe adequately not only oxygen dissolved evolution, but also of the production of xanthan and substrate consumption. Also, the influence of several parameters (k(L)a, air flow rate and dissolved oxygen) in the evolution of the key compounds of the system have been studied. The results of the simulation shown that an increasing of dissolved oxygen concentration favor the xanthan gum production. PMID- 11064051 TI - At-line monitoring of a submerged filamentous bacterial cultivation using near infrared spectroscopy. AB - The use of near infra-red spectroscopy (NIRS) to monitor a submerged filamentous bacterial bioprocess was investigated. An industrial strain of the filamentous bacterium Streptomyces fradiae was cultured in a 12 litre stirred tank reactor (STR) using a complex medium. This mycelial 4 phase (oil, water, gas and solid) system produced highly complex and variable matrices, therefore monitoring such a complex fluid with NIRS represented a considerable challenge. Nevertheless, successful models for four key analytes (methyl oleate, glucose, glutamate and ammonium) were built at-line (rapid off-line) using NIRS. In the present study, the methods used to formulate, select and validate the models for the key analytes are discussed, with particular emphasis on how the model performance can be critically evaluated. Since previous reports on NIRS in monitoring bioprocesses have either involved simpler matrices, or, in filamentous systems, have not discussed how NIRS models can be critically assessed, the emphasis in the present study on providing an insight into the modelling process in such a complex matrix, may be particularly important to the applicability of NIRS to such industrial bioprocesses. PMID- 11064052 TI - Influence of glycerol production on the aerobic and anaerobic growth of the wine yeast Candida stellata. AB - Candida stellata is frequently found in wine fermentations and may be used as a yeast starter in beverage production. In order to acquire additional knowledge on the physiology of C. stellata, a study on sugar metabolism in aerobic and anaerobic conditions was carried out. We found that under anaerobic conditions the low growth rate and biomass yield of C. stellata were due to the diversion of carbon flux from ethanol to glycerol. C. stellata had lower ADHI (alcohol dehydrogenase) activity (3-4 fold) and higher GPDH (glycerol-3-phosphate dehydrogenase) activity (40 and 15 times higher in anaerobiosis and aerobiosis respectively) than that of a Saccharomyces cerevisiae control strain. In aerobic sugar-limited chemostat culture C. stellata exhibited lower maximum biomass concentration [5.23 gl(-1) (dry weight)] than other respirofermentative yeasts at very low dilution rates (up to D = 0.042 h(-1)). While glycerol was constantly produced, ethanol and sugar residue appeared at D = 0.042 h(-1) and D = 0.065 h( 1) respectively. The tendency of C. stellata to form glycerol is probably the main cause of its very low growth and fermentation rates. PMID- 11064053 TI - Addition of polar organic solvents can improve the product selectivity of cyclodextrin glycosyltransferase. Solvent effects on cgtase. AB - Cyclodextrin glycosyltransferase (EC 2.4.1.19, CGTase) is an enzyme that produces cyclodextrins from starch via an intramolecular transglycosylation reaction. Addition of small amounts (10% v/v) of polar organic solvents can affect both the overall production yield and the type of cyclodextrin produced from a maltodextrin substrate under simulated industrial process conditions. Using CGTase from Thermoanaerobacter sp. all solvents produced an increase in cyclodextrin yield when compared with a control, the greatest increase being obtained with addition of ethanol (26%). In addition product selectivity was affected by the nature of the organic solvent used: beta-cyclodextrin was favoured in the absence of any solvent and on the addition of dimethylsulphoxide, t-butanol and dimethylformanide while alpha-cyclodextrin was favoured by addition of acetonitrile, ethanol and tetrahydrofuran. With CGTase from Bacillus circulans strain 251 relatively smaller increases in overall cyclodextrin production were achieved (between 5-10%). Addition of t-butanol to a B. circulans catalysed reaction however did produce the largest selectivity for beta-cyclodextrin of any solvent-enzyme combination (82%). The effect of solvent addition was shown not to be related to the product inhibition of CGTase, but may be related to reduced competition from the intermolecular transglycosylation reaction that causes degradation of cyclodextrin products. This rate of this reaction was shown to be dependent on the nature of the organic solvent used. PMID- 11064054 TI - Hydrocarbon degradation by a soil microbial population with beta-cyclodextrin as surfactant to enhance bioavailability. AB - In general the biodegradation of nonchlorinated aliphatic and aromatic hydrocarbons is influenced by their bioavailability. Hydrocarbons are very poorly soluble in water. They are easily adsorbed to clay or humus fractions in the soil, and pass very slowly to the aqueous phase, where they are metabolised by microorganisms. Surfactants that increase their solubility and improve their bioavailability can thereby accelerate degradation. Cyclodextrins are natural compounds that form soluble complexes with hydrophobic molecules. They are widely used in medicine and harmless to microorganisms and enzymes. This paper describes their in vitro effect on the biodegradative activity of a microbial population isolated from a petroleum-polluted soil, as shown by the decrease of dodecane (C12), tetracosane (C24) anthracene and naphthalene added individually as the sole carbon source to mineral medium liquid cultures. beta-cyclodextrin accelerated the degradation of all four hydrocarbons, particularly naphthalene, and influenced the growth kinetics as shown by a higher biomass yield and better utilization of hydrocarbon as a carbon and energy source. Its low cost, biocompatibility and effective acceleration of degradation make beta-cyclodextrin an attractive option for bioremediation. PMID- 11064055 TI - Scale-up study on suspension cultures of Taxus chinensis cells for production of taxane diterpene. AB - Suspension cells of Taxus chinensis were cultivated in both shake flasks and bioreactors. The production of taxuyunnanine C (TC) was greatly reduced when the cell cultures were transferred from shake flasks to bioreactors. Oxygen supply, shear stress and stripping-off of gaseous metabolites were considered as potential factors affecting the taxane accumulation in bioreactors. The effects of oxygen supply on the cell growth and metabolism were investigated in a stirred tank bioreactor by altering its oxygen transfer rate (OTR). It was found that both the pattern and amount of TC accumulation were not much changed within the range of OTR as investigated. Comparative studies on the cell cultivation in low shear and high shear generating bioreactors suggest that the decrease of TC formation in bioreactors was not due to the different shear environments in different cultivation vessels. An incorporation of 2% CO(2) in the inlet air was beneficial for the cell growth, but did not improve the TC production in bioreactors. Furthermore, the effects of different levels of ethylene addition into the inlet air on the cell growth and TC production were investigated in a bubble column reactor. The average cell growth rate increased from 0.146 to 0.204 d(-1) as the ethylene concentration was raised from 0 to 50 ppm, and both the content and production of TC were also greatly improved by ethylene addition. At an ethylene concentration of 18 ppm, the highest TC content and volumetric production in the reactor reached 13.28 mg/(g DW) and 163.7 mg/L, respectively, which were almost the same as those in shake flasks. Compared with the control reactor (bubble column without ethylene supplementation), the maximum TC content was increased by 82% and the total production of TC was doubled. The results indicate that ethylene is a key factor in scaling up the process of the suspension cultures of T. chinensis from a shake flask to a bioreactor. PMID- 11064056 TI - Tough love. PMID- 11064058 TI - Anal sphincter replacement(1). PMID- 11064057 TI - Repairing adult inguinal hernias: let us count the ways(1). PMID- 11064059 TI - Physician profiling: is it a reliable measure of quality of care?(1). PMID- 11064060 TI - Intrauterine repair of myelomeningoceles(1). PMID- 11064061 TI - Hepatitis C: the real danger to surgeons(1). PMID- 11064062 TI - Achalasia: looking for the best route(1). PMID- 11064063 TI - Approaches to the organ donor shortage(1). PMID- 11064065 TI - Primary repair of penetrating colonic injuries(1). PMID- 11064064 TI - Pediatric liver transplantation: trends in liver transplantation in children(1). PMID- 11064066 TI - Enterocutaneous fistulas: current diagnosis and management. PMID- 11064067 TI - Grand rounds in general surgery. PMID- 11064068 TI - Early recognition of hepatic portal vein gas on CT with appropriate surgical intervention improves patient survival. AB - Historically, hepatic portal vein gas has been linked with a dismal prognosis. However, in the last 3 decades, identification of hepatic portal vein gas has undergone a transition from late recognition on roentgenograms to earlier identification on computed tomography scan. The 5 patients in this case series were found to harbor hepatic portal vein gas identified using computed tomography scan. Eighty percent of the patients had concomitant pathology identified by computed tomography, felt to be the cause of the hepatic portal vein gas. All patients were taken to the operating room for exploration and resection of the offending pathology. All 5 patients survived. We suggest that early recognition using computed tomography scan, with appropriate operative intervention, improves the chance for patient survival when hepatic portal vein gas is identified. PMID- 11064069 TI - Evaluation of a disease management plan for prevention and diagnosis of thromboembolic disease in major trauma patients. AB - Evaluate the effectiveness of our program of thromboembolic disease (TED) prophylaxis.Multisystem trauma patients with closed head injury, spinal cord injury, and/or long bone/pelvic fractures severe enough to require inpatient rehabilitation are at high risk for TED: deep venous thrombosis (DVT) and pulmonary embolism (PE). A retrospective analysis of 262 such trauma patients, from September 1995 through July 1997, was performed to evaluate the effectiveness of our program of TED prophylaxis. Mechanical compression devices (CDs) and/or anticoagulation treatment were the primary methods of prophylaxis. If clinical suspicion of TED was present, duplex ultrasound (DU) was used. Otherwise, patients were screened once by DU before transfer to a rehabilitation facility.There was 99% compliance with prophylaxis use. One hundred one patients died of injuries less than 48 hours after admission. Forty-four patients died in the hospital more than 48 hours after admission; 4 (9%) had TED manifest as DVTs (3 patients; 66% at the level of the popliteal or above) and PE (1 patient; 2%). One hundred seventeen patients survived and were transferred to rehabilitation facilities. Fourteen of these (12%) had TED manifest as DVTs (13 patients; 71% at the level of the popliteal or above) before transfer and PE (1 patient; 1%). Three patients developed DVTs without PEs during 2-month follow-up in rehabilitation. Of the 14 patients who developed TED, 7 had prophylaxis with CDs alone and 5 had CDs plus heparin, aspirin, or caval filter (CD+), an incidence of 11%. Increased injury severity score and increased length of stay were associated with development of TED (p < 0.05). Forty-two percent of patients with TED were suspected on the basis of clinical examination and confirmed by DU; 9 of 105 (8.5%) were found exclusively by the single DU screening of clinically negative lower extremities.The compliance rate for TED prophylaxis in these severely injured trauma patients was high. An 11% incidence of TED was found in this population; 4 of 160 patients (2.5%) died with or from TED. Compression devices alone or CD+ offered equal prophylaxis. Single DU screening of clinically negative lower extremities demonstrated an 8.6% incidence of TED; clinical suspicion of TED as an indication for DU screening increased positive findings to 42%. PMID- 11064070 TI - General surgery program directors' perceptions of the match. AB - To identify expectations of general surgery program directors (PDs) for recruitment behavior, and to document the experiences, perceptions, and ethical dilemmas they experienced with the 1998 National Resident Matching Program (NRMP).Two hundred sixty-five general surgery PDs were asked to complete a 30 item written questionnaire. Questions inquired about PD perceptions of students' interviewing practices, how communication with applicants is conducted and interpreted, and overall perceptions of the match.A total of 77.7% of PDs responded. A total of 44% of PDs found nothing wrong with students interviewing in multiple specialties and conceded legitimate reasons for doing so; yet, 75% of PDs felt this practice negatively affected students' rank order. A total of 46.6% of programs told students to keep in touch to be ranked; only 8.7% of PDs believed students' stated interest, and 76.6% of PDs said these affirmations had no effect on students' rank. A total of 36.5% of PDs felt students made informal commitments to them, and 90.4% felt students at least sometimes lied to them. A total of 51.7% of PDs felt the match was a reasonable process that needed no changes.As long as the stakes are high and there are no repercussions for unethical behaviors and practices during residency recruitment, gamesmanship will continue to be the accepted culture. PMID- 11064071 TI - The effects of acute coronary occlusion on noninvasive echocardiographically derived systolic and diastolic myocardial strain rates. AB - To compare the magnitude and rates of change of peak systolic (epsilon(')(SYS)) and diastolic (epsilon(')(DIAS)) strain rates, as measured using tissue Doppler echocardiography, to pressure and volumes left-ventricular indices during acute coronary occlusion.Six closed-chest dogs had a combination high-fidelity conductance pressure transducer placed into the left ventricle for determination of end-diastolic and end-systolic pressures (EDP and ESP) and volumes (EDV, ESV, and ejection fraction [EF]). Other indices included the time constant of left ventricular relaxation (tau), +dP/dt(max), -dP/dt(max), end-systolic pressure/volume index (ESPV). A coronary angioplasty catheter was positioned into the left-anterior descending coronary artery. During coronary occlusion, strain rates and hemodynamic parameters were recorded continuously for 2 minutes.During occlusion, significant decreases in strain rates occurred within 30 seconds. Systolic indices (ESPV and +dP/dt(max)) changed at rates similar to epsilon(')(SYS) (each p = NS). Diastolic indices (tau, EDP, EDV, -dp/dt(max)) also changed at rates similar to epsilon(')(DIAS) (each p = NS). However, EF decreased at a significantly slower rate than did strain rates (p < 0.05).CONCLUSIONS:Peak systolic and diastolic strain rates decrease with acute ischemia similar to corresponding indices of left-ventricular systolic and diastolic function. Strain rates may be used in the noninvasive assessment of ischemic-induced left-ventricular dysfunction. PMID- 11064072 TI - Amebic abscess of the liver presenting as acute cholecystitis. AB - A case report is presented of a 37-year-old active duty Navy petty officer with amebic abscess of the liver presenting as acute cholecystitis. He was admitted with severe right upper quadrant pain and a positive Murphy's sign, but sonogram and computed tomography (CT) scan demonstrated an abscess in the right lobe of the liver. "Anchovy paste" material was obtained on percutaneous drainage, and he was placed on metronidazole administration with a tentative diagnosis of amebic abscess. This was confirmed on enzyme-linked immunosorbent assay. Symptoms resolved within a few days, and the abscess progressively decreased in size. Amebic abscess of the liver is discussed, with emphasis on pathogenesis, diagnosis, and treatment. Although uncommon, the condition is still seen in various population groups including the United States military. Difficulty in diagnosis is not unusual, and as in the herein-reported case, amebic abscesses of the liver may be confused with acute cholecystitis and other intra-abdominal infections. Abdominal sonogram and CT examination will identify a process in the liver, but the presence of amebiasis must be confirmed by laboratory studies on serum or contents of the abscess. Amebicidal agents are effective in many cases, but there remain roles for aspiration of the abscess, percutaneous drainage, and even open surgical drainage. Failure to establish an early diagnosis may result in rupture of the abscess, with catastrophic results. PMID- 11064073 TI - Delaying the skin for TRAM flaps. AB - This study was undertaken to determine whether a less extensive delay procedure would be as efficacious as the standard delay procedure in breast reconstruction.Between July 1996 and February 1999, 15 patients underwent delay procedures prior to breast reconstruction. Six patients underwent the standard delay procedure. Nine patients underwent a less extensive skin delay procedure. Transverse rectus abdominis myocutaneous (TRAM) reconstruction was performed 1 week after delay procedures.Average operating time was 28.7 minutes for the standard delay and 19.7 minutes for the skin paddle delay. The incidences of fat necrosis were 17% in the standard delay group and 22% in the skin paddle delay group. The incidences of partial flap loss/slow healing were 17% in the standard delay group and 22% in the skin paddle delay group. The incidence of complications in each group was the same: approximately 1 per patient.Operating times were not statistically different between the two groups (p = 0.06). There was no increase in the incidence of slow healing/partial flap loss or fat necrosis in the skin delay group. The skin delay procedure for TRAM flaps seems to provide a concise delay procedure that does not increase the incidence of complications in those high-risk patients. PMID- 11064074 TI - Extrauterine pregnancy: a historical review(3). AB - This article reviews historical developments in the recognition and management of abdominal and ectopic pregnancy since these entities were first recognized centuries ago. Contributions of the early anatomists who studied the female reproductive system and the courage of early surgeons who dared invade the abdomen cleared the path for the extraordinary success of today's obstetricians and gynecologists in the treatment of conditions formerly fatal to both mother and fetus. PMID- 11064075 TI - Impact of blood transfusion on outcome in patients admitted for gastrointestinal hemorrhage. AB - PURPOSE:Patients admitted with the diagnosis of gastrointestinal bleeding at our institution typically undergo diagnostic/therapeutic endoscopy. Surgery is consulted and operative intervention considered when the patient has reached a 6 unit transfusion requirement for resuscitation. The purpose of this study is to examine the association between number of units transfused and clinical outcome in patients admitted for gastrointestinal hemorrhage.A retrospective review of records of patients admitted to the 81st Medical Group between January 1996 and January 2000 was conducted. Patients admitted with a diagnosis of upper or lower gastrointestinal hemorrhage were identified. Of this group, the records of those patients receiving at least 1 unit of packed red blood cells were examined.Thirty five complete records were available for review. Patients ranged from 3 to 79 years of age. The male to female ratio was 4:1. The group received an average of 5.2 units of packed red blood cells. Eight patients were identified as having received more than 6 units of packed red cells. Three of 4 patients who underwent operative intervention had transfusion requirements in excess of 6 units. All 4 operative patients were classified as ASA class IIIE. Seven deaths (20%) occurred among the 35 patients, 3 of whom had received more than 6 units of blood.Patients admitted with gastrointestinal hemorrhage who require less than 6 units of blood may have a lower chance of dying (15%) than do patients requiring more than 6 units of blood (38%). Emergent surgical intervention, even in high-risk patients, can be safely performed. PMID- 11064076 TI - Torsion of an intra-abdominal testis. AB - To present a case of torsion of a nonneoplastic intra-abdominal testis with an unusual clinical presentation.A 26-year-old active duty Navy Petty Officer presented to the emergency department on 3 occasions over a 5-day period with lower abdominal pain. Physical examination demonstrated acute tenderness in the left lower quadrant with sugestion of a normal spermatic cord and atrophic testis in the left scrotum. Computed tomography scan demonstrated an intra-abdominal lesion near the internal inguinal ring. The patient underwent surgical exploration through an inguinal incision. Torsion of a nonviable intra-abdominal testis was present. The scrotum contained only the vas deferens and cremasteric muscle. An orchiectomy was performed with removal of the vas deferens and other cord structures.The unusual clinical finding of acute torsion of an intra abdominal testis, associated with an apparent atrophic scrotal testis, presented a confusing clinical picture. Computed tomography scan did not clarify the issue sufficiently to establish a definite preoperative diagnosis. Clinical suspicion prompted early surgical intervention. Review of the current literature produced 60 reported cases of torsion of an intra-abdominal testis. Two thirds of these involved testicular neoplasm, usually seminoma. Although the clinical presentation varied, most patients had recent onset of lower abdominal pain associated with tenderness and, in half the cases, a mass. Patients almost always presented with an absent scrotal testis on the involved side, and not infrequently reported previous surgery thought to be an orchiectomy.Diagnosis of an intra-abdominal testicular torsion is rare, particularly when no neoplasm is present. A high index of suspicion must be maintained whenever there is abdominal pain and undescended testis. The surgical history and imaging studies may not clarify a confusing clinical picture. PMID- 11064077 TI - Enteral feeding tubes in the intensive care unit: efficiency of placement and morbidities associated with use. PMID- 11064078 TI - Glucagonlike peptide 2 (glp-2) promotes intestinal recovery following chemotherapy-induced enteritis. PMID- 11064079 TI - Akt and prostate tumorigenesis. PMID- 11064080 TI - Does age affect outcome of gastric bypass surgery? PMID- 11064081 TI - Outcome following combined carotid endarterectomy and coronary artery bypass is related to patient selection. PMID- 11064082 TI - Transposed basilic vein fistula: a superior alternative to prosthetic grafts? PMID- 11064083 TI - The development of a novel spinal cord nerve guide with an injectable trophic factor delivery system. PMID- 11064084 TI - Do glutamine-supplemented enteral feedings improve outcome in critically ill patients? PMID- 11064085 TI - Interval hepatic resection of colorectal metastases improves patient selection* PMID- 11064086 TI - Acute scrotal swelling: unusual presentation of perforated sigmoid diverticulitis. PMID- 11064087 TI - Virus infection abrogates cd8(+) t cell deletion induced by donor-specific transfusion and anti-cd154 monoclonal antibody. PMID- 11064088 TI - In vitro study of vascular endothelial growth factor for bioengineering improved prosthetic vascular grafts. PMID- 11064089 TI - Discharge after cardiac surgery: where do patients go and why? PMID- 11064090 TI - The increased prevalence of hypothyroidism in women newly diagnosed with breast carcinoma at st. mary's hospital* PMID- 11064091 TI - Laparoscopic versus open appendectomy. PMID- 11064092 TI - Is routine duplex ultrasound surveillance after carotid endarterectomy cost effective? PMID- 11064093 TI - The use of hyperbaric oxygen for the management of calciphylaxis. PMID- 11064094 TI - Intravenous pyelography is unnecessary for the preoperative evaluation of living renal donors* PMID- 11064095 TI - In vivo characterization of the molecular-genetic changes during the development of acute stress-associated gastric ulceration* PMID- 11064096 TI - Applications of electrosurgery: radio frequency ablation of liver tumors. PMID- 11064097 TI - Immune reconstitution: an important component of a successful allogeneic transplantation. AB - The recipients of allogeneic haematopoietic stem cell transplants are characterised by an immunodeficiency of varying severity and duration. Their immunocompromised state is due in part to: (1) an impaired recapitulation of lymphoid ontogeny, (2) a lack of sustained transfer of donor immunity, (3) the effects of graft versus host disease and its therapy, and (4) a reduction in thymic function. Recipients can have delays in the production of naive T lymphocytes following transplantation which result in defects in the production of new antigen specific T lymphocytes and an inability to produce antibodies, especially to carbohydrate antigens. T-cell proliferation as well as immunoglobulin production remains impaired usually until the second half of the first year post-transplant. Other factors that can influence immunological reconstitution include the donor-recipient relationship (histocompatible or matched unrelated donor), intervening infections and recipient age, among others. PMID- 11064098 TI - Systemic or local co-administration of lactoferrin with sensitizing dose of antigen enhances delayed type hypersensitivity in mice. AB - Lactoferrin (LF), a major defense protein synthesized and stored in granulocytes has been implicated in maintaining immune homeostasis during an insult-induced metabolic imbalance. In this study, we demonstrated that lactoferrin augments the delayed type hypersensitivity (DTH) response to specific antigens in mice. Lactoferrin (LF) was given to mice orally or intraperitoneally (i.p. ) at the time of immunization, or subcutaneously (s.c.) in a mixture with the immunizing doses of the following antigens, sheep red blood cells (SRBC), Calmette-Guerin bacillus (BCG) or ovalbumin (OVA). A DTH reaction was determined 24 h after administration of an eliciting dose of antigen as a specific increase in foot pad swelling. Lactoferrin enhanced DTH reaction to all studied antigens in a dose dependent manner. Lactoferrin (LF) given to mice in conjunction with antigen administered in an incomplete Freund's adjuvant induced the DTH response at the level of control mice given antigen in a complete Freund's adjuvant. In addition, LF remarkably increased DTH response to a very small, otherwise non-immunogenic SRBC dose. The increase in DTH response was less pronounced for orally administered LF than for any other routes of administration, however, statistically significant augmentation was demonstrated for each antigen studied. Although the costimulatory action of LF was accompanied by the appearance of bovine lactoferrin-specific cellular responses in mice, it is very unlikely that such responses will be generated in humans, since bovine lactoferrin is a dietary antigen to which a tolerance has been acquired. Considering the involvement of LF in generation of stimulatory signals during the induction phase of an antigen specific immune responses, we suggest that LF may be useful for development of safer and more efficacious vaccination protocols. PMID- 11064099 TI - CXCR4 and CCR5 expression by H9 T-cells is downregulated by a peptide-nucleic acid immunomodulator. AB - Product R (Reticulose(TM)) is a peptide-nucleic acid immunomodulator with broad spectrum antiviral activity that was recently shown to increase expression of mRNAs encoding the proinflammatory cytokines, IFN-gamma, IL-1beta, IL-6 and TNF alpha. Since these cytokines induce expression of the chemokines, MIP-1alpha, MIP 1beta, RANTES, and SDF-1, all of which inhibit viral infectivity, we were interested to determine if Product R also alters chemokine expression. In addition, the finding, that Product R decreases HIV-1 RNA and extracellular p24 antigen in H9 T-lymphoma cells, suggested to us that this drug may block viral infection by reducing the expression of chemokine receptors on target cells. We have therefore utilized H9 cells to test the effects of Product R on expression of mRNAs encoding the chemokine receptors, CD4, CXCR4 and CCR5, as well as their ligands, IL-16, SDF-1, MIP-1alpha, MIP-1beta, and RANTES, by RT-PCR. We also assayed the effect of Product R on surface receptor expression by flow cytometry, and on the chemotactic activity of these cells towards the CXCR4 ligand, SDF-1, and the CCR5 ligands, MIP-1alpha and RANTES. H9 cells were cultured for 3-21 days in medium containing 5% or 10% Product R, or 5% or 10% PBS. We found that, compared to control cultures, cells cultured in media containing Product R expressed lower amounts of CXCR4 and CCR5 mRNA and surface antigen at all time points. Culture for 3 days in media containing Product R also reduced the ability of cells to migrate towards 10-20 ng/ml SDF-1 and 100-250 ng/ml RANTES. In contrast, Product R had no effect on the expression of CD4 mRNA and receptor protein, or on expression of IL-16 mRNA. These findings suggest that Product R may have clinical efficacy in HIV-1-infected patients by downregulating viral coreceptors on target T-cells. PMID- 11064100 TI - The quantitative characteristics of efficiency of ballistic transfection of chimeric antibody genes. AB - A quantitative approach was applied to the study of in vivo expression of foreign genes introduced into mice by ballistic transfection. Because in some cases one must take into account both the level of synthesized protein and that of antibodies to it, we derived the equation which allows to calculate the exact quantity of both proteins. This formula was applied to in vivo expression of a chimeric (human/mice) immunoglobulin E gene. The immunochemical analysis using this equation showed that the Ig concentration succeeded 4, 6, 12 IU/ml and undetectable level, respectively, upon transfection in mouse liver, spleen, foot pad and ear cartilage. PMID- 11064101 TI - Differences in the changes of allergen-specific IgE serum levels and the chemiluminescence of peripheral blood phagocytes in patients with allergic rhinoconjunctivitis during the ragweed season. AB - The objective of this study was to compare the changes in the values of allergen specific serum IgE levels and zymosan-induced whole blood chemiluminescence (CL) in 41 patients who had exclusively only ragweed allergy in the season of acute symptoms of disease in July, August and September. All patients had allergic rhinitis or rhinoconjunctivitis. Each patient was investigated as a self-control. The ragweed-specific IgE levels were measured by enzyme immunoassay (EIA). The luminol amplified zymosan-induced CL of whole human blood was detected. The allergen-specific serum IgE levels showed slight, but not significant, gradually increasing elevations during the whole season. On the other hand, significant increases were found in the values of the basal but especially in the zymosan stimulated CL of peripheral blood phagocytes during the acute phase of allergy. Both the basal and the zymosan-induced CL reflected significantly the activated state of the immune system. These observations clearly show that there are well detectable signs of the systemic activation of the immune system in allergic rhinoconjunctivitis beside the local alterations. In addition, the measurements of the basal and zymosan-induced CL of peripheral phagocytes could clearly reflect the clinical state of disease in vitro. PMID- 11064102 TI - Measurement of intracellular interferon-gamma and interleukin-4 in whole blood T lymphocytes from patients with systemic lupus erythematosus. AB - Contradictory data are available about the dominance of T-helper 1 (T(H)1), or T helper 2 (T(H)2) cytokines in systemic lupus erythematosus (SLE). Therefore, intracellular interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production of T lymphocytes was measured in whole blood of healthy donors and active and inactive SLE patients by flow cytometry. The percentage of IFN-gamma and IL-4 positive cells was low (<1%) in unstimulated samples of the healthy controls, while that of IFN-gamma and IL-4 positive cells in the stimulated cells was 25.2+/-10.6% and 0.6+/-1.5%, respectively. No significant difference was found between SLE patients and healthy controls and between active and inactive patients in these parameters either in the unstimulated or in the stimulated samples. One patient with severe disease had as high as 11.8% IL-4 positive cells and 12.5% IFN-gamma positive cells in the stimulated samples, but after the initiation of intensive corticosteroid and cytostatic therapy, the percentage of IL-4 positive T cells decreased (4.76%) while that of IFN-gamma positive T cells increased (47.91%). We conclude that the intracellular IL-4 and IFN-gamma expression of T lymphocytes does not differ markedly between SLE patients and healthy controls, with the possible exception of severe disease, when marked IL-4 overproduction may exist beside low IFN-gamma production. Furthermore, corticosteroid and cytostatic therapy might normalize this altered IFN-gamma/IL-4 ratio. PMID- 11064103 TI - Atomic force microscopy to study direct neurite-mast cell (RBL) communication in vitro. AB - Communication between nerves and mast cells is a prototypic demonstration of neuroimmune interaction. We used an in vitro co-culture approach comprising cultured murine superior cervical ganglia (SCG) and rat basophilic leukemia (RBL 2H3) cells. Atomic force microscopy (AFM) showed how neurites attached to a pseudopodium or a cell body of an RBL cell. After stimulation of SCG neurites with bradykinin or scorpion venom, RBL cells attached to neurites spread and flattened, and several discharged granules (0. 5-1.0 microm in diameter) were found on the surface of the RBL cells. A neurokinin (NK)-1 receptor (i.e. substance P receptor) antagonist prevented the RBL degranulation. The results showed that activation of the SCG neurites with bradykinin or scorpion venom was able to elicit degranulation in RBL cells which were attached to neurites. PMID- 11064104 TI - Naked DNA when co-administered intranasally with heat-labile enterotoxin of Escherichia coli primes effectively for systemic B- and T-cell responses to the encoded antigen. AB - In this study a novel prime-boost immunisation strategy was evaluated. Priming of BALB/c mice by the intranasal route with plasmid DNA encoding beta-galactosidase (LacZ) with or without heat-labile enterotoxin (LT) of Escherichia coli as a mucosal adjuvant, resulted in the induction of weak serum antibody and proliferative T-cell responses. However, following an intraperitoneal booster injection with the beta-galactosidase protein (beta-gal), strong antibody and proliferative T-cell responses were induced in all the mice. These responses were highest in mice primed intranasally with a mixture of LacZ+LT as compared to those mice primed with DNA (LacZ) or protein (beta-gal) alone. Moreover, LacZ+LT primed mice produced high avidity antibodies and the subclasses of serum antibodies were IgG1 and IgG2a, suggesting a mixed Th1/Th2-type response. Priming of mice with either protein (beta-gal) or DNA (LacZ) alone, produced predominantly IgG1 antibodies, suggesting a Th2-type response. These findings suggest that the use of a heterologous DNA-prime, protein-boost immunisation scheme combining different routes of administration, might be an advantageous strategy for the induction of accelerated immune responses. PMID- 11064105 TI - Identification of human homologue of mouse IFN-gamma induced protein from human dendritic cells. PMID- 11064106 TI - L-selectin gene T668C mutation in type 1 diabetes patients and their first degree relatives. AB - There have been some studies published recently which have suggested that L selectin and/or other adhesion molecules could be the new markers for diabetes type 1 risk development in humans and animal models of the disease. The alterations of soluble L-selectin have been found not only in overt but also in the preclinical stage of disease development and were independent from the presence of ICA - a marker of ongoing autoimmunity, but associated with HLA related genetic predisposition to insulin-dependent diabetes mellitus (IDDM). The aim of our study was to evaluate the frequency of the L-selectin gene T668C mutation (from thymine to cytosine at position 668) resulted in F206L an amino acid substitution in patients with overt diabetes and their unaffected first degree relatives in comparison to the unselected control population. In the unaffected siblings of IDDM subjects we have observed a significantly higher frequency of the L-selectin gene T668C mutation in comparison to their relatives with type 1 diabetes and healthy controls. It was also shown that there is an association between T668C mutation and low HLA related risk of IDDM development, the highest frequency of F206L mutation in the EGF domain of L-selectin was observed in relatives with 'protective' HLA DQB1*0602 allele and nonDRB1*03 nonDRB1*04 haplotype, while in subjects with highest risk of IDDM haplotype the frequency of T668C mutation was similar to the controls. We would like to hypothesise that the T668C L-selectin gene mutation could have a (protective?) role in the development of IDDM, but further studies concerning their role in type 1 diabetes are needed. PMID- 11064107 TI - Serological HLA class I alleles in Senegalese blood donors detected HBs Ag positive. AB - We analysed the HLA class I alleles in 96 blood donors HBs Ag positive compared with 93 healthy control individuals (HBs negative). The most frequent HLA-A, -B, C alleles found were, A23 (33.6%); A2 (25%); A30 (25%); B8 (31.5%); B7 (16.3%); B58 (11.9%); B35 (11.9%); B49 (11.9%); B53 (10.8%); Cw7 (39.1%); Cw3 (36.9%); Cw4 (36.9%). Significant differences (P<0.001) were found between the blood donors and the controls for the following HLA alleles, A1; A23; B8 and Cw3. The detection of HBe antigen was positive in 26/84 blood donors. It was observed a significant difference (P<0.01; odds ratios (OR)=6.25) between positive and negative HBe antigens blood donors for HLA-A1 allele. PMID- 11064108 TI - Induction of active systemic anaphylaxis by oral sensitization with ovalbumin in mast-cell-deficient mice. AB - Mast-cell-deficient W/W(v) mice were sensitized by oral administration of 0.1 and 1.0 mg ovalbumin (OVA) by gavage every day for 9 weeks, and active systemic anaphylaxis (ASA) was induced by intraperitoneal injection of OVA. The production of OVA-specific IgE and IgG1 by oral immunization of the W/W(v) mice was high, and the production of IL-4 by splenocytes re-stimulated with OVA in vitro was increased. In contrast, production of OVA-specific IgG2a and IgG2b was low, and production of IFN-gamma by splenocytes after re-stimulation with OVA in vitro was rather decreased. These findings suggest that Th2-dominant helper T-cell activation had occurred. No increase in serum histamine level was observed following ASA induction. However, the plasma platelet-activating factor (PAF) levels of the mice sensitized with 0.1 and 1.0 mg OVA by gavage increased significantly. The increases in plasma PAF correlated well with the ASA associated decreases in body temperature, suggesting that PAF plays an important role in ASA in W/W(v) mice. Taken together the above findings indicate that W/W(v) mice are a good model not only for studying induction of food allergy but also for examining the role of PAF in food-induced hypersensitivity. PMID- 11064109 TI - Innate immune response mechanisms in non-insulin dependent diabetes mellitus patients assessed by flow cytoenzymology. AB - It is well known that infections in patients with diabetes mellitus are more severe, although there is controversy for increased susceptibility to them. Non specific immune response mechanisms could be related to defense and/or susceptibility to pathogens. The aim of this study was to investigate the activity of several enzymes involved in the primary host defense mechanisms in non-insulin dependent diabetes mellitus (NIDDM). Twenty NIDDM females with a mean HbA(1c) level of 8.19% were included. No patient had clinical evidence of infection. As controls 20 healthy females were studied. The enzymes tested were dipeptidyl-peptidase I (DPP-I), cathepsin B and D, NADPH oxidase and superoxide dismutase (oxidative burst) and collagenase. Isolated leukocytes were incubated with the specific substrates in pyrogen free conditions. The intracellular enzyme activity was analyzed by flow cytometry. Collagenase enzymatic activity was similar in the three leukocyte subpopulations studied. Oxidative burst induction in monocytes was comparable between both groups. Enzyme activity of cathepsin B and D in all cell subsets, oxidative burst in PMN cells, and DPP-I in lymphocytes and monocytes from patients, was higher than those from healthy females (P<0.05). Overall, our findings demonstrate an enhanced functional status of several intracellular leukocyte enzymes in NIDDM. Furthermore, the increased oxidative burst induction and the consequent production of free radicals, may contribute to vascular complications. Other mechanisms - either from the non-specific or specific immune response - deserve investigation to establish if diabetic patients are more susceptible to infectious diseases. PMID- 11064110 TI - Strain-dependent antibody response induced by DNA immunization. AB - The antibody (Ab) response induced by DNA-based immunization was compared in various strains of inbred, H-2 congenic and outbred mice with different haplotypes of mouse major histocompatibility complex (H-2). Two different plasmid expression vectors encoding Aequorea victoria green fluorescent protein (GFP) or Escherichia coli, beta-galactosidase (beta-gal) were introduced into quadriceps muscle, and Ab production was examined using both enzyme-linked immunosorbent assay and immunoblot analysis. The beta-gal plasmid DNA immunization induced strong Ab production in all inbred, H-2 congenic and outbred strains at the early stages of immunization. By comparison with beta-gal peptide immunization, the degree of Ab response was H-2 haplotype-dependent. On the other hand, Ab production by GFP plasmid DNA immunization was observed in outbred strains, but not in some of the inbred and H-2 congenic strains. Also, outbred strains showed a high Ab response compared with other inbred and H-2 congenic strains by GFP peptide immunization. Reverse transcription-polymerase chain reaction analysis demonstrated the presence of GFP or beta-gal transcripts at the DNA inoculation site in all the strains studied, even in inbred and H-2 congenic strains which showed no Ab production by GFP plasmid DNA immunization. These results indicate that the difference in Ab response induced by DNA immunization as well as by peptide immunization depends upon the H-2 haplotypes of host strains. PMID- 11064111 TI - Rapid small-scale isolation of SV40 virions and SV40 DNA. AB - A rapid method for the small-scale isolation of SV40 virions and SV40 DNA is presented. CV-1 monkey epithelial cells are transfected with linear SV40 DNA. After the onset of transfection, cells are lysed by several freeze/thaw cycles and virions are isolated using polyethylene glycol (PEG) precipitation of DNase I treated lysates. Viral DNA is released by proteinase K and dithiothreitol treatment of the isolated virions followed by phenol/chloroform extraction and ethanol precipitation. This method yields on average 7.5x10(4) plaque forming units (PFUs) and DNA of adequate purity and concentration to be used for restriction analysis on ethidium bromide agarose gels from a single 35-mm tissue culture dish. PMID- 11064112 TI - Design and validation of immunological tests for the detection of Porcine endogenous retrovirus in biological materials. AB - The present study details the design and demonstrates function for a series of reagents and methods to allow the detection of exposure to antigens specific for Porcine endogenous retrovirus (PERV). The detection of PERV is carried out by the means of a variety of immunological screening methods including, indirect immunofluorescence, Western blotting and enzyme linked immunosorbent assay (ELISA) for the detection of antibodies in serum specific for PERV gag and env antigens. Alternatively, PERV-specific antisera for gag and env can be used to detect viral antigen in serum or other samples. PERV env peptides with potential specificity for the known PERV types are also described. Antisera against the peptides can be used to detect PERV antigens directly or to characterise viral type. Using electron microscopy coupled with labelled PERV-gag-specific antisera it was possible to visualise PERV virions. PMID- 11064113 TI - A microtitre plate method for isolation and typing of poliovirus using a blue cell ELISA. AB - A simple, sensitive, specific and rapid procedure for isolating and typing polioviruses is described. Specimens are inoculated onto confluent monolayers of cell lines (Hep-2C, L20B or RD) seeded into microtitre plates. After 24-48 h, the infected cells are stained with monoclonal antibodies specific for poliovirus types 1,2,3 or a blend of the three antibodies followed by an anti-mouse IgG horseradish peroxidase conjugate. On addition of substrate, infected cells stain an intense blue colour and are easily distinguished from uninfected cells by light microscopy. Poliovirus infection can be detected before the appearance of cytopathic effects (CPE). This Blue-Cell ELISA test was evaluated against conventional culture and seroneutralisation on a range of polio isolates and clinical specimens. The sensitivity and specificity of the Blue-Cell ELISA compared to neutralisation was 100% (87/87) on culture supernatants of poliovirus isolates sent to our reference laboratory for confirmation. All the poliovirus isolates were typed within 24 h of specimen inoculation using the new method compared to 6-10 days by conventional culture and neutralisation. The method proved to be more sensitive than conventional culture when clinical specimens were examined. Of 43 clinical specimens from which poliovirus had been previously isolated by various laboratories in the U.K., 30/43 (69.8%) were positive for poliovirus by the Blue-Cell ELISA compared to 29/43 (67.4%) by conventional culture and neutralisation. Neutralisation of specimens exhibiting CPE indicated that all of the polioviruses were correctly typed with the new method. CPE was not observed by conventional culture in any specimen that was negative in the Blue-Cell ELISA. There were no cross-reactions with a range of other enteroviruses. PMID- 11064114 TI - Comparative analysis of TMV-Cg and TMV-U1 detection methods in infected Arabidopsis thaliana. AB - The common strain of the tobacco mosaic virus (TMV-U1), and the crucifer infecting tobacco mosaic virus (TMV-Cg), both members of Tobamovirus genus, infect efficiently the solanaceous plants such as tomato and tobacco. The crucifer-infecting tobacco mosaic virus (TMV-Cg) also infects Arabidopsis thaliana plant, spreading systemically without causing severe symptoms. In contrast, Arabidopsis is a poor host for TMV-U1 infection. Within the past 10 years, Arabidopsis has developed into a powerful model system for studying plant pathogen interaction. However, a detailed analysis comparing the accuracy of various viral detection methods has not been reported previously. Four detection methods were evaluated in A. thaliana (ecotype Po-1), infected with TMV-U1 or TMV Cg. Western blots, enzyme-linked immunosorbent assay (ELISA), reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ RNA hybridization methods were used to determine viral spread at various days post inoculation (dpi) in inoculated and apical non-inoculated leaves. The detection of viral spread of TMV-U1 and TMV-Cg in Arabidopsis, using these four detection methods, supports previous studies, which demonstrate that the systemic spreads of these two viruses differ in Arabidopsis. Western blotting and ELISA detected TMV-Cg at 5dpi, and TMV-U1 at 12 dpi in systemic tissues. Viral spread was detected earlier when using RNA detection methods. Reverse transcriptase-polymerase chain reaction (RT-PCR) was very sensitive for detecting TMV-Cg in A. thaliana, but less sensitive for TMV-U1 detection. In situ RNA hybridization showed differential distribution of TMV-Cg and TMV-U1 in the inoculated leaf and systemic tissues. PMID- 11064115 TI - Detection of parvovirus B19 IgM by antibody capture enzyme immunoassay: receiver operating characteristic analysis. AB - Parvovirus B19 infection can cause severe effects in high-risk groups including pregnant women and immunocompromised individuals. Although serological detection of B19 infection is commonplace, minimal information is available on the absolute performance characteristics of various tests for the detection of B19 IgM. The performance of the first parvovirus B19 IgM enzyme immunoassay to be cleared by the US Food and Drug Administration (FDA) is described. The immunoassay cut-off has been established using receiver operating characteristic (ROC) analysis giving a sensitivity and specificity of detection of 89.1 and 99.4%, respectively. No cross-reactivity is observed with rubella or other viral disease IgM which cause similar symptomologies to parvovirus B19. Multi-site reproducibility studies have shown high immunoassay reproducibility with detection rates (observed/expected result) of 100% for nonreactive specimens (N=324) and strongly reactive (N=403), respectively. Immunoassay reproducibility ranged from 11.76 to 17. 46% coefficient of variation for all reactive specimens tested (N=12) whereby each specimen was assayed a total of 81 times. Parvovirus B19 IgM seroprevalence of 1% was observed in a US blood donor population (N=399). In the absence of international performance criteria, this study will be of major benefit to the clinical virologist in assessing immunoassay reliability for the detection of recent infection with parvovirus B19. PMID- 11064116 TI - Shortening of the diagnostic window with a new combined HIV p24 antigen and anti HIV-1/2/O screening test. AB - Because antibodies to the human immunodeficiency virus (HIV) are absent in the very early phase of HIV infection, there remains a slight residual risk for HIV transmission by blood donations by viremic but antibody negative donations. To shorten the diagnostic window between infection and the detection of antibodies, Enzygnost HIV Integral (Dade Behring, Germany) was developed. With this new test, HIV p24 antigen and HIV antibodies can be detected simultaneously in a single test. In a multicenter study the new screening assay has been compared with various tests that detect only HIV antibodies or HIV p24 antigen and with assays which permit a simultaneous detection of HIV antigen and HIV antibodies. The new assay showed 100% sensitivity for the detection of antibodies to HIV-1, groups M (n=1102) and O (n=55), and HIV-2 (n=289). In 23 out of 52 seroconversion panels, seroconversion was detected 2-18 days earlier with the new combined antigen/antibody test compared to single antibody tests. All samples from a viral load panel (n=451), all samples containing p24 antigen (n=302), and all but one of the cell culture supernatants (n=38) infected with various HIV-1 subtypes or HIV-2 were identified reliably by the new test. The specificity of the assay for 4002 unselected blood donors was 99.78% initially and 99.80% after retesting. Potentially interfering factors had no systematic influence on specificity. By testing for p24 antigen, which is present prior to the onset of antibody production in some cases of recent HIV infection, the new assay reduces the diagnostic window as compared to third generation screening assays, thus permitting an earlier diagnosis of HIV infection. PMID- 11064117 TI - Development and evaluation of ELISA procedures to detect antibodies against the major envelope protein (G(L)) of equine arteritis virus. AB - Enzyme-linked immunosorbant assays (ELISAs) were developed for the detection of antibodies against the major envelope glycoprotein (G(L)) of equine arteritis virus (EAV). A 6-Histidine tagged recombinant protein expressing the complete G(L) ectodomain (G(L)-6His), a glutathione-S-transferase recombinant protein expressing amino acids 55-98 of G(L) (G(L)-GST) and an ovalbumin-conjugated synthetic peptide representing amino acids 81-106 of G(L) (G(L)-OVA) were used as diagnostic antigens. An ELISA procedure was developed and optimised for each antigen. The G(L)-OVA and G(L)-6His assays showed the greatest specificity while the G(L)-GST assay was slightly more sensitive that the G(L)-OVA and G(L)-6His assays; results based on the analysis of 50 virus neutralisation positive and 50 virus neutralisation negative sera. The G(L)-OVA ELISA was selected for further evaluation since it was simpler to use than ELISAs based on recombinant antigens and did not suffer from background reactivity. The final sensitivity and specificity of the G(L)-OVA ELISA were 96.75 and 95.6%, respectively, results based on the analysis of 400 virus neutralisation positive and 400 virus neutralisation negative sera. It also detected EAV antibody (100% efficiency) in seropositive shedding stallions and, in ponies infected experimentally with the UK93 isolate of EAV, the appearance of virus neutralising antibodies and G(L)-OVA ELISA-specific immunoglobulins coincided. PMID- 11064118 TI - Comparison of assays to detect cytomegalovirus shedding in the semen of HIV infected men. AB - We sought to determine the optimal assays for cytomegalovirus (CMV) shedding in semen. Over a 2-month period, 149 HIV-1-infected men who have sex with men each provided up to three semen specimens. Specimens were tested for CMV by culture, rapid assay (shell vial) and polymerase chain reaction (PCR). By culture, 30% of seminal plasma and 28% of seminal cell specimens grew CMV. By rapid assay, results were 38 and 33%, respectively. By PCR, 56% of seminal cell specimens demonstrated CMV: 20% in a single semen specimen; 33% in two specimens; and 34% in all three specimens. Overall, 69% of men had CMV detected by PCR in at least one seminal cell specimen. By quantitative PCR, 14% had ten, 14% had 100, 16% had 1000, and 12% had 10000 copies in 6.25 microl of semen analyzed. Adjusting for initial CD4+ cell count, men with CMV shedding demonstrated by PCR at the first visit were approximately four times as likely to shed CMV at a subsequent visit (RR 4.28, CI: 2.30-7.95). CMV shedding was associated with decreased CD4+ cell counts in peripheral blood (P=0.05). It is concluded that the PCR assay provided the greatest sensitivity among the three detection methods. PMID- 11064119 TI - Production monitoring and purification of EBV encoded latent membrane protein 1 expressed and secreted by recombinant baculovirus infected insect cells. AB - Epstein-Barr virus (EBV) encoded latent membrane protein 1 (LMP1) is expressed in malignancies with latency type II and III and is an important transforming protein. To further study this protein LMP1 was expressed by and purified from recombinant baculovirus infected Sf9 cells. Expression levels of LMP1 in EBV transformed B cell lines and Sf9 cells were analyzed using a newly developed quantitative LMP1-capture ELISA. Highest expression was found in the cell line X50/7 (6.2 ng/10(7) cells), whereas expression levels of recombinant LMP1 (bLMP1) in Sf9 cells reached 506 ng/10(7) cells. Surprisingly bLMP1 could also be detected in the culture medium as a stable full-length protein. Highest expression in Sf9 cells (506 ng/10(7) cells) was observed at 48 h post infection and in the culture medium (1590 ng/ml) at 96 h post infection. Before purification bLMP1 was solubilised using 0.22 m octyl-beta-glucoside at pH 6.0. Purification of bLMP1 using Q-Sepharose FF yielded 10-80 times enriched bLMP1 fractions, indicating that Q-Sepharose can be used for pre-purification. A one step monoclonal antibody based immunoaffinity chromatography yielded highly purified bLMP1. Although the overall yields (20 microg purified LMP1 from 100 ml culture supernatant) and protein concentrations were low, higher concentrations of >95% purified BLMP1 could be reached after freeze drying. PMID- 11064120 TI - Rapid diagnosis and quantification of herpes simplex virus with a green fluorescent protein reporter system. AB - A genetically modified cell line (Vero-ICP10-EGFP) was constructed for detection of herpes simplex virus (HSV) by a simple, rapid and direct method. The cell line was developed by stable transfection of Vero cell with a plasmid encoding the green fluorescent protein (GFP) driven by the promoter of the HSV-2 ICP10 gene. As early as 6 h after infection with HSV, fluorescence-emitting cells can be observed under a fluorescence microscope. A single infected cell emitting fluorescence can be observed with soft agar overlay by inverted fluorescence microscopy. No induction of detectable fluorescence was seen following infections with human cytomegalovirus (HCMV), varicella zoster virus (VZV), coxsackievirus A16 and enterovirus 71. Analysis by flow cytometry also demonstrated that intensity of the triggered fluorescence is proportional to the titer of HSV inoculated. Taken together, this novel GFP reporter system could become a useful means for rapid detection and quantification of HSV in clinical specimens. PMID- 11064121 TI - Identification of novel cytochrome P450 1A genes from five marine mammal species. AB - Marine mammals, being endangered by the chronic exposure of hydrophobic environmental contaminants as an assorting result of global pollution, are especially focused as indicators for organochlorine pollution. The use of contaminant-induced xenobiotic metabolizers, particularly P450 (CYP) 1A, in marine mammals can be effective as potential biomarkers of the contaminant exposure and/or toxic effects. In this study, we identified the first marine mammalian CYPs. Six novel CYP1A cDNA fragments were cloned from the livers of marine mammal species, minke whale (Balaenoptera acutorostrata), dall's porpoise (Phocoenoides dalli), steller sea lion (Eumetopias jubatus), largha seal (Phoca largha), and ribbon seal (Phoca fasciata) by the method of reverse transcription/polymerase chain reaction (RT/PCR); two distinct fragments were from steller sea lion and one fragment each was obtained from the other species. Five of the fragments, one from each species, were classified in the subfamily of CYP1A1, and the other fragment cloned from steller sea lion was designated CYP1A2. Degenerate PCR primers were used to amplify the fragments from liver cDNAs. The deduced amino acid sequences of these fragment CYP1As showed identities ranging from 50.0 to 94.3% with other known vertebrate CYPs in the subfamily of CYP1A, including those from fish, chicken, and terrestrial mammals. The isolated fragments were used to construct a molecular phylogeny, along with other vertebrate CYP1A cDNAs cut down in size to the corresponding region of 265 bp in which those newly determined fragments were cloned. This phylogenetic analysis by the maximum parsimony method using the PHYLIP program suggests two distinct evolutional pathways for aquatic mammalian CYP1As, compatible to a conservative taxonomy. Pinniped genes are clustered together with dog gene, forming a carnivore group, and cetaceans form another branch. Identification of CYP1A genes in marine mammals will be an introductory step to provide new insights into the metabolic or toxicological functions of CYP1As in these animals. PMID- 11064122 TI - Valve movement responses of Velesunio angasi (Bivalvia: Hyriidae) to manganese and uranium: an exception to the free ion activity model. AB - The veracity of the free ion activity model (FIAM) was tested by examining the acute (48 h) valve movement responses (VMR) (measured in terms of the duration of valve opening) of the Australian tropical freshwater unionid bivalve, Velesunio angasi to increasing concentrations of total Mn or U, in a standard synthetic water under conditions of varying pH (5.0-6.0) and/or dissolved organic carbon (model fulvic acid, FA) concentrations (0-8.9 mg l(-1)). Valve movement behaviour, measured using an automated data acquisition system, was shown to be a quantifiable and rapid, real-time endpoint for assessing the toxic effects of Mn and U exposures. For Mn, the VMR of V. angasi were independent (P>0.05) of pH and/or model FA concentration. In contrast, VMR to U exposures were highly dependent (P< or =0.05) on pH and/or model FA concentration; individuals were more sensitive to U at low pH and model FA concentrations. Valve movement responses to Mn were directly proportional to the activity of the free metal ion (Mn(2+)), which is consistent with the FIAM. In contrast, VMR to U were regarded as an 'exception' to the FIAM, since they were a weighted function of the activities of the free metal ion and the 1:1 metal hydroxide species (i.e. 1.86 x UO2(2+) + UO2OH(+)). Additionally, the effect of U on V. angasi demonstrates the importance of examining VMR at more than one pH. At a fixed pH, the results for U were consistent with the FIAM (i.e. response was directly proportional to UO2(2+)); only when pH was altered, were the results inconsistent with the FIAM. The inconsistency in the VMR of V. angasi to U exposures in this study, together with similar examples from other studies using different metals (e.g. Al or Zn), raises questions regarding the veracity of the FIAM. A detailed examination of the conceptual development of the FIAM is required to probe its apparent failure to describe several metal-organism interactions. PMID- 11064123 TI - Evaluation of the free ion activity model of metal-organism interaction: extension of the conceptual model. AB - The present study integrates the concepts of the free ion activity model (FIAM) into biological receptor theory (BRT; i.e. pharmacodynamic principles) to obtain a more rigorous conceptual model; one that more precisely quantifies the interaction of chemical species at biological receptor sites. The developed model, which is viewed as an extended FIAM, explains the conditions under which the FIAM will be effective in explaining biological response (BR). It establishes that BR is directly proportional to the activity of the free metal ion in the linear regions of concentration-response curves only. Additionally, it indicates that [X-cell], the activity of free surface sites on the cell membrane, does not need to be constant in the region of BR, as assumed by the original FIAM. The extended FIAM was tested by re-examining concentration-response data from the literature on aquatic organisms exposed to several ecotoxicologically-relevant trace metals. These data, which would be considered exceptions to the original FIAM, were found to be consistent with the extended FIAM. Due to its more rigorous conceptual basis, the extended FIAM is capable of modelling concentration-response experiments from a wider range of water chemistry conditions (i.e. varying pH, hardness and dissolved organic matter) than the original model and, as such, potentially provides a more useful tool for evaluating metal-organism interactions. This study proposes, for the first time, a quantitative method of uncoupling the biological effects of a metal hydroxide (1:1) complex from that of amelioration of the free metal ion (M(z+)) by H(+). Since the activities of H(+) and metal-hydroxide cannot be independently varied, it has been previously very difficult to evaluate whether metal-hydroxide species contribute to eliciting a BR. Furthermore, the extended FIAM can directly derive fundamental information from concentration-response curves, such as the binding constants of H(+) or the hardness cations (Ca(2+) and/or Mg(2+)) to the cell membrane surface of aquatic organisms. PMID- 11064124 TI - Effects of an acute silver challenge on survival, silver distribution and ionoregulation within developing rainbow trout eggs (Oncorhynchus mykiss). AB - Rainbow trout eggs were acutely challenged with silver (as AgNO(3)) at different stages of development from fertilization through to hatch in moderately hard water (120 mg CaCO(3) l(-1), 0.70 mM (25 mg l(-1)) Cl(-), 1.3 mg l(-1) DOC, 12.3+/-0.1 degrees C) at measured total silver concentrations of 0.11+/-0.004, 1.55+/-0.15, and 14.15+/-1.52 microg l(-1). Four separate acute challenges were conducted, each consisting of 5 days exposure to the respective silver concentration, followed by 4 days recovery after transfer to silver-free water (series 1, 1-10 days post-fertilization; series 2, 8-17 days post-fertilization; series 3, 16-25 days post-fertilization; series 4, 23-32 days post fertilization). Mortality was not significantly different from control during exposure to 0.11, 1.55, and 14.15 microg l(-1) total silver in series 2, 3 and 4 (mortality for series 1 data could not be calculated for technical reasons). In the four days of recovery following silver exposure, however, there was significant mortality at 14.15 microg l(-1) total silver reaching 100, 31 and 72% in series 2, 3 and 4, respectively, indicating eggs are more sensitive in the period of 8-17 and 23-32 days post-fertilization at this temperature. Mortality following silver exposure was associated with ionoregulatory impairment in series 3 and 4, where up to 60% of whole egg [Na(+)] and [Cl(-)] was lost relative to controls at 14.15 microg l(-1) total silver. Significant but smaller reductions in egg [Na(+)] and/or [Cl(-)] were also observed at 0.11 and 1.55 microg l(-1) total silver. The greatest accumulation of silver in whole eggs and chorions occurred in series 4, reaching concentrations of 0.53 microg g(-1) (eggs) and 15.5 microg g(-1) (chorions) in the 14.15 microg l(-1) treatment. The accumulation of silver in the whole eggs and chorions of the 0.11 microg l(-1) treatment was not different from controls throughout embryonic development. Of the total silver content, only a small proportion of silver was found in the embryos (1-17%), indicating that the chorion is a protective barrier during acute silver exposure. PMID- 11064125 TI - Effects of pre-acclimation to aluminium on the physiology and swimming behaviour of juvenile rainbow trout (Oncorhynchus mykiss) during a pulsed exposure. AB - Anthropogenic acidification of the freshwater environment causes aluminium to be mobilised into the aquatic environment. When pH falls below 5.5, exposure to aluminium concentrations as low as 12.5 microg.l(-1) can cause serious physiological disturbances in freshwater fish. However, under constant laboratory exposures fish can acclimate and recover physiological status within 5-30 days. In reality, fish in the wild are likely to experience chronic sub-lethal exposure, with occasional elevations (pulses) to much higher levels. The experiment described here investigated the effects of an environmentally realistic, 4-day pulse exposure to a high level of aluminium (36 microg.l(-1)) in two groups of juvenile rainbow trout. One group was exposed to a lower level of aluminium (24 microg.l(-1)) for 16 days before and 10 days after the pulse ('aluminium-acclimated' fish). A second group was exposed to pH 5.2 alone for 16 days before and 10 days after the pulse ('aluminium-naive' fish). A third group exposed to pH 5.2 alone for 30 days (no aluminium added) acted as controls. Triplicate groups of 24 juvenile rainbow trout (2.3-16.7 g) were randomly allocated to one of these three treatments. Swimming behaviour was monitored throughout and samples were taken on days 14, 20, 22, 26 and 30 for assessment of physiological status. No treatment effects were recorded in the control group (pH 5.2 alone). Fish in the 'aluminium-acclimated' treatment became hypo-active upon initiation of the exposure to 24 microg.l(-1) aluminium, but recovered after just 4 days of this exposure. Subsequent challenge on day 16 with the 36 microg.l(-1) aluminium 'pulse' caused these fish to became hypo-active again, but they recovered normal swimming behaviour whilst still subject to the 4-day pulse. The 'aluminium-naive' fish also became hypo-active during the pulse exposure (36 microg.l(-1) aluminium). However, they did not exhibit any recovery of swimming behaviour, either during the pulse, or even 6 days after the cessation of the pulse, despite a rapid depuration of gill aluminium load (within 2 days of the pulse finishing). Mortality was low in the aluminium-acclimated fish (4%) and significantly higher in the aluminium-naive fish (26%). Haematological disturbances were most extreme in the aluminium-naive fish and had not recovered to control levels 6 days after the end of the pulse. This study provides new evidence, using behavioural responses, that previous exposure to low levels of aluminium may be an important factor abating the impact of aluminium on fish in the natural environment. PMID- 11064126 TI - The effects of diffusible creosote-derived compounds on development in Pacific herring (Clupea pallasi). AB - The effects of diffusible creosote-derived compounds from weathered creosote treated pilings on embryonic development in the Pacific herring were investigated. Parameters used to evaluate toxicity included embryonic development, cardiac function, embryo/larval activity (movement of developing embryos), hatching success, and larval morphology at hatch. For acute exposures, embryos were incubated in seawater containing either creosote-treated wood (creosote) or untreated wood (wood control), or seawater alone (control). All embryos adhering directly to creosote-treated wood and 40-50% of embryos not adhering to the creosote-treated wood failed to develop beyond the first few days of incubation. For surviving embryos, a 93% reduction in heart rate, and moderate to marked arrhythmia was observed. Surviving embryos also exhibited both an increase in frequency and an alteration in pattern of embryo/larval movement, with most embryos exhibiting tremors as compared with the vigorous movements of the control embryos. Cardiac function and embryo/larval movements of embryos exposed to untreated wood were not significantly different from controls. The hatching rate of embryos exposed to creosote was 90% lower than control embryos and 72.4% lower than embryos exposed to untreated wood, and the LC(50) for hatching success was 0.05 mg/l. Partial hatching (incomplete hatch) was observed in 15-20% of embryos exposed to creosote. All of the hatched larvae exposed as embryos to creosote exhibited morphological deformities, including scoliosis, pericardial edema and/or ascites. Similar effects were observed in embryos collected from creosoted pilings in San Francisco Bay, with a 72% decrease in hatching success compared with embryos collected from the Bay and severely deformed larvae. To investigate the combined effects of creosote and salinity on hatching success, larval morphology, and cardiac function, embryos were exposed to a sublethal concentration of creosote (0.003 mg/l) at three salinities; sub optimal (8 parts per thousand (ppt)), optimal (16 ppt), and high salinity (28 ppt). The presence of creosote decreased hatching success at all three salinities, but the effect was greatest at 8 ppt (34% reduction) and the least in 28 ppt (14% reduction). The increased incidence of morphological abnormalities was also smallest at the high salinity (10% compared with 24 and 33% in 8 and 16 ppt). While exposure to creosote resulted in reduced heart rates at all three salinities, no additive effect of creosote and salinity was observed. PMID- 11064127 TI - PAH metabolites in bile, cytochrome P4501A and DNA adducts as environmental risk parameters for chronic oil exposure: a laboratory experiment with Atlantic cod. AB - In order to perform environmental risk assessments with regard to oil contamination in the sea, it is important to obtain knowledge about threshold levels for possible adverse effects in marine organisms. With this objective in mind, selected biomarkers were studied in Atlantic cod (Gadus morhua) chronically exposed to mechanically dispersed crude oil. The fish were exposed for 30 days in a continuous flow system to nominal concentrations of 0.06, 0.25 and 1 ppm North Sea crude oil. Fish were sampled five times during the exposure period. In addition, the 1 ppm group and the control group were sampled 1 week after the end of exposure. Polyaromatic hydrocarbon (PAH) concentrations in the seawater were analysed regularly by direct fluorescence and, at one occasion, by gas chromatography with mass spectrographic detection (GC/MS) measurements. Liver samples were analysed for parent PAH levels by means of GC/MS measurements, and PAH metabolites in bile were analysed by means of fixed wavelength fluorescence. Cytochrome P450 induction in liver was estimated by ethoxyresorufin-O-deethylase (EROD) activity, and hepatic DNA adducts were analysed by the 32P-postlabelling assay. The parent PAH concentrations in liver showed peak levels 3 days after the start of exposure, followed by a reduction towards the end of the experiment. In contrast, the PAH metabolites in bile and EROD activity showed generally increasing levels throughout the whole exposure period, indicating an increased biotransformation efficiency. The level of DNA adducts in the 1 ppm group showed a stable increase during the entire exposure period. Only a slight, non significant decrease in DNA adduct levels was observed after 7 days of recovery in clean water. Exposure-dependent responses were observed for all three biomarkers. The lowest nominal concentration of dispersed oil in water, 0.06 ppm, corresponded to a measured total PAH concentration in the water of 0.3 ppb. Atlantic cod exposed to this concentration showed increased levels of PAH metabolites in bile and a slight induction of CYP1A, as well as formation of DNA adducts when compared with control fish. Particularly noteworthy is the detection of DNA adducts at such a low exposure concentration of oil in water, which, to our knowledge, is a novel finding. These dose-response data may serve as useful contributions when assessing environmental risk with regard to marine oil pollution. PMID- 11064128 TI - Exposure of brown trout, Salmo trutta, to a sub-lethal concentration of copper in soft acidic water: effects upon muscle metabolism and membrane potential. AB - Brown trout acclimated to soft water and exposed for 96 h to a sub-lethal concentration of copper at low pH (0.08 micromol l(-1) Cu, pH 5) have a lower critical swimming speed than fish from copper-free water at neutral pH. This loss of performance is not due to difficulties in oxygen transfer resulting from gill damage since arterial oxygen and carbon dioxide levels remain unaffected. Both red and white muscle showed some metabolic disruptions consistent with local hypoxia, namely a high lactate concentration at rest and, in the white muscle, depletion of glycogen and phosphocreatine. However, a putative role of increased blood viscosity following haematological changes in reducing the supply of oxygen to the tissues is not supported by the current study. Haematocrit, haemoglobin and plasma protein concentrations were not affected by this treatment and a lack of further change in variables such as lactate at the onset of exercise led one to look for an alternative explanation for the effects of copper and low pH upon tissue metabolites. Ammonia concentration, both in the plasma and muscles, is significantly higher in trout exposed to copper and low pH. Ammonia plays a role in the regulation of a number of metabolic pathways and could contribute to the altered metabolic status of these fish. In addition, ammonium ions are known to cause electrophysiological disruptions, particularly the displacement of K(+) in ion exchange mechanisms that could lead to the observed loss of swimming performance. Using the measured distribution of ammonia between intracellular and extracellular compartments to estimate membrane potential of resting muscle, a significant depolarisation is predicted in both red and white muscle of fish exposed to copper and low pH. PMID- 11064130 TI - Self-assembled hydrogel nanoparticles from curdlan derivatives: characterization, anti-cancer drug release and interaction with a hepatoma cell line (HepG2). AB - Self-assembled hydrogel nanoparticles were synthesized from carboxymethylated (CM)-curdlan, substituted with a sulfonylurea (SU) as a hydrophobic moiety for self-assembly. The degree of SU substitution was 2.4, 5.6, or 7.2 SU groups per hundred anhydroglucose units of curdlan. The physicochemical properties of the self-assembled hydrogel nanoparticles (DS 2.4, DS 5.6, and DS 7.2) in aqueous media were characterized by dynamic light scattering, transmission electron microscopy, and fluorescence spectroscopy. The mean diameter of all samples was less than 300 nm with a unimodal size distribution. The critical aggregation concentrations (CAC) of self-assembled hydrogel nanoparticles in distilled water were 4.2 x 10(-2), 3.1 x 10(-2) and 1.9 x 10(-2) mg/ml for DS 2.4, 5.6 and 7.2, respectively. The loading and release of all-trans retinoic acid (ATRA) was studied. The ATRA loading efficiencies and loading contents of CM-curdlan/SU nanoparticles increased as the degree of SU substitution increased. The ATRA release rate was controlled by the degree of substitution and drug-loading. For specific interaction with a hepatic carcinoma cell line (HepG2), CM-curdlan was additionally conjugated with lactobionic acid (LBA; galactose moiety) (5.5 LBA molecules per hundred glucose units). HepG2 was strongly luminated by ligand receptor interactions with fluorescence-labeled LBA/CM-curdlan/SU hydrogel nanoparticles. The luminescence was not observed for other control cases. It is concluded that LBA/CM-curdlan/SU hydrogel nanoparticles are a useful drug carrier for the treatment of liver cancer, because of the potential immunological enhancement activities of CM-curdlan in the body, the ligand-receptor mediated specific interactions, and the controlled release of the anti-cancer drug. PMID- 11064131 TI - Mucoadhesive drug carriers based on complexes of poly(acrylic acid) and PEGylated drugs having hydrolysable PEG-anhydride-drug linkages. AB - We have designed a new mucoadhesive drug delivery formulation based on H-bonded complexes of poly(acrylic acid) (PAA) or poly(methacrylic acid) (PMAA) with the poly(ethylene glycol) (PEG), of a (PEG)-drug conjugate. The PEGylated prodrugs are synthesized with degradable PEG-anhydride-drug bonds for eventual delivery of free drug from the formulation. In this work we have used indomethacin as the model drug which is PEGylated via anhydride bonds to the PEG. The complexes are designed first to dissociate as the formulation swells in contact with mucosal surfaces at pH 7.4, releasing PEG-indomethacin, which then hydrolyses to release free drug and free PEG. We found that as MW of PAA increases, the dissociation rate of the complex decreases, which results in decreased rate of release of the drug. On the other hand, the drug release from PEG-indomethacin alone and from solid mixture of PEG-indomethacin+PAA was much faster than that from the H-bonded complexes. Due to the differences in the thermal stability, PMAA complex exhibited slightly faster drug release than that of the PAA complex of comparable MW. These H-bonded complexes of degradable PEGylated drugs with bioadhesive polymers should be useful for mucosal drug delivery. PMID- 11064132 TI - Biological potency of microsphere encapsulated plasmid DNA. AB - We evaluated the utility of three in vitro methods to monitor the biological potency of PLGA encapsulated DNA. For each assay we also determined whether the biological activity was influenced by the structural profile of DNA isomers. Collectively, the results indicate that all three methods can be used to evaluate the biological activity of DNA extracted from PLGA microspheres, but they are differentially sensitive to the structural changes of plasmid DNA that can occur during microencapsulation and microsphere storage. More specifically, mammalian cell transfection followed by an enzyme assay affords an accurate determination of DNA potency over time and is less influenced by DNA isoform than bacterial transformation. Cell-free transcription/translation systems can also be utilized, and the results of this assay are influenced by DNA isoform. Finally, bacterial transformation was found to be more sensitive to DNA isoform than the other assay methods. PMID- 11064133 TI - Mechanistic aspects of the release of levamisole hydrochloride from biodegradable polymers. AB - The release of levamisole hydrochloride from poly-DL-lactide-co-glycolide compacts prepared at 5, 10 and 20% drug loading using two different particle size fractions of drug (90-125 and 125-250 microm) was investigated. Release profiles were significantly different from those previously reported for compacts prepared using the base form of the drug. Release was found to occur in a biphasic manner, with an initial fast release phase followed by a slower polymer degradation controlled release phase. The drug release profiles were successfully described by a model combining contributions from a first-order initial release phase and a polymer degradation controlled drug release phase. The fraction of drug released in the initial burst phase (F(B)) was attributed to the dissolution of drug domains situated at the surface of the polymer-drug compact and this fraction tended to increase with increasing drug particle size, as expected from the model. The increase in F(B) with increased loading was attributed to the clumping of dispersed drug particles which effectively increased the proportion of drug linked to the compact surface. PMID- 11064134 TI - FTIR-ATR spectroscopy for monitoring polyanhydride/anhydride-amine reactions. AB - The reactivity of 1,3-bis(p-carboxyphenoxy) propane:sebacic acid anhydride copolymer (CPPSA1:6), myristic and benzoic anhydrides with amine nucleophiles were investigated in non-polar solvents. FTIR-ATR (attenuated total reflectance) spectroscopy was used to monitor the polyanhydride/anhydride reaction rates in dichloromethane, dichloroethane, chloroform, and 1,4-dioxane solutions at room temperature. The reaction kinetics was determined by measuring the anhydride peak loss with time. Aminolysis resulted from nucleophilic attack of the added amine on the carbonyl group of the anhydride moiety. Primary and secondary amines reacted to form amides and the reaction followed second-order kinetics. Second order rate constants and reaction half-life (t(1/2)) were calculated from the semilog plots of [anhydride]/[amine] in 1,4-dioxane at room temperature. The aminolysis rate decreased with pK(a) of the amine reactant, and half-life (t(1/2)) decreased with increasing amine concentration, as expected. With trifluoroethylamine (pK(a) 5.8), myristic anhydride reacted about 6-fold faster than benzoic anhydride. The lower reaction rate of benzoic anhydride was due to the higher stability of the aromatic anhydride compared to aliphatic. The overall CPPSA1:6 copolymer reactivity was the sum of aliphatic-aliphatic (SA-SA), aliphatic-aromatic (SA-CPP), and aromatic-aromatic (CPP-CPP) anhydride linkage reactivities. Based on the monomer ratio, the probability of SA-SA, SA-CPP, and CPP-CPP dyads were calculated to be 0.74, 0.24, and 0.02, respectively. This indicated that CPPSA1:6 reactivity will mainly result from SA-SA and SA-CPP linkages. The second-order rate constants and t(1/2) obtained for CPPSA1:6 with TFEA were closer to those for myristic anhydride than benzoic anhydride with TFEA. PMID- 11064135 TI - Influence of the co-encapsulation of different non-ionic surfactants on the properties of PLGA insulin-loaded microspheres. AB - The aim of this work was to produce insulin-loaded microspheres allowing the preservation of peptide stability during both particle processing and insulin release. Our strategy was to combine the concepts of using surfactants to improve insulin stability while optimising overall microsphere characteristics such as size, morphology, peptide loading and release. Bovine insulin was encapsulated within poly(lactide-co-glycolide) (PLGA 50:50, Resomer RG504H) microspheres by the multiple emulsion-solvent evaporation technique. Microspheres were prepared by adding to the primary emulsion three non-ionic surfactants, poloxamer 188, polysorbate 20 and sorbitan monooleate 80, at different concentrations (1.5 and 3. 0% w/v). The presence of surfactants was found to decrease the mean diameter and to affect the morphology of the microspheres. Insulin encapsulation efficiency was reduced in the presence of surfactants and especially for sorbitan monooleate 80, in a concentration-dependent mode. The influence of the surfactants on the interactions between insulin and PLGA together with the primary emulsion stability were found to be the major determinants of insulin encapsulation. The release of insulin from microspheres was biphasic, showing an initial burst effect followed by a near zero-order release for all the batches prepared. The initial burst was related to the presence of insulin molecules located onto or near to the microsphere surface. In the presence of surfactants, a faster insulin release with respect to microspheres encapsulating insulin alone was observed. Insulin stability within microspheres after processing, storage and release was evaluated by reversed phase- and size-exclusion-HPLC. The analysis of microsphere content after processing and 6 months of storage showed that insulin did not undergo any chemical modification within microspheres. On the contrary, during the period of sustained release insulin was transformed in a high molecular weight product, the amount of which was related to the surfactant used. In conclusion, polysorbate 20 at 3% w/v concentration was the most effective in giving regular shaped particles with both good insulin loading and slow release, and limiting insulin modification within microspheres. PMID- 11064136 TI - pH-responsive pseudo-peptides for cell membrane disruption. AB - We describe pseudo-peptides obtained by the copolymerisation of L-lysine and L lysine ethyl-ester with various hydrophobic dicarboxylic acid moieties. In aqueous solution, when the carboxylic acid groups are charged, the polymers dissolve. When they are fully neutralised the hydrophobic moieties cause the polymer to precipitate. The pH range over which reversible precipitation occurs can be adjusted by changing the intramolecular hydrophilic/hydrophobic balance, by using a carboxylic acid moiety with a different pK(a) value or by changing the apparent pK(a) value of the polymer through chemical modifications of the backbone. These bio-degradable materials are well tolerated by a range of mammalian cell lines at physiological pH but display an ability to associate with the outer membranes of these cells, which they rupture to varying degrees at pH 5.5. Relative to the degree of lysis displayed by poly(L-lysine iso-phthalamide), lysis was reduced by partial esterification and increased by replacing the aromatic iso-phthaloyl moiety with a long chain aliphatic dodecyl moiety. Similar behaviour was observed for the pH-dependent rupture of human erythrocytes, where poly(L-lysine dodecanamide) displayed enhanced cell lysis at pH values <7.0 relative to poly(L-lysine iso-phthalamide). PMID- 11064137 TI - Characterization of commercially available and synthesized polyethylenimines for gene delivery. AB - Five new polyethylenimines (PEI) were synthesized by polymerization of aziridine in aqueous solution and compared to several commercially available PEI used for gene transfer. Polymers were characterized by 13C NMR spectroscopy, capillary viscosimetry, potentiometric titration and Cu(II) complex formation to gain insight into structural and functional properties. 13C NMR analysis revealed differences in the extent of branching based on the ratio of primary, secondary and tertiary amino groups. An amino group ratio 1 degrees :2 degrees :3 degrees =1:2:1 was obtained for the synthesized PEI, whereas commercially available PEI generally showed a higher degree of branching (1:1:1). Capillary viscosimetry of aqueous PEI solutions with a sufficient amount of salt gave Mark-Houwink parameters of alpha=0.26 and K(V)=1.00 cm(3)/g for the commercially available polymers. In case of the synthesized polymers, variation of reaction conditions yielded viscosity average molar masses (M(v)) in the range of 8000-24000 g/mol. PEI solutions were investigated by potentiometric titration analysis showing that their buffer capacity was not significantly influenced by molar mass or polymer structure. The pK(a) values (8.18-9.94) and the buffer capacity beta (0.08-0.014 mol/l) were of comparable magnitude. This study highlights the necessity of more detailed characterization methods for PEI used in gene transfer protocols since physico-chemical properties do not reflect the vast differences found in transfection efficiencies. PMID- 11064138 TI - Improved monitoring for brain dysfunction in intensive care and surgery: introduction PMID- 11064139 TI - IBIS data library: clinical description of the Finnish database. Improved Monitoring for Brain Dysfunction during Intensive Care and Surgery. AB - Improved monitoring of brain function in intensive care and surgery is a project aiming to develop methods of biosignal processing and interpretation, in order to characterise critical events during anaesthesia and cardiac surgery, effect of anaesthesia regimen, neurophysiological findings of different sedation levels, arousal from post-anaesthesia sedation, post-cardiopulmonary bypass brain dysfunction and early brain dysfunction in patients with multiple organ failure. A data library was collected in the three participating hospitals. This is a description of the data library from the University Hospital of Kuopio, Finland, which includes data from 40 patients after cardiac surgery and from seven patients with multiple organ dysfunction. This project demonstrates that active neuromonitoring can be performed in the intensive care unit without interference with the normal treatment and care. The presented database may serve other scientific workers as a reference for a typical spectrum of perioperative data with respect to severity of disease, length of cardiopulmonary bypass, postoperative levels of sedation and length of hospital stay in cardiac surgery patients, and for types of diseases and outcome in patients with multiple organ dysfunction. PMID- 11064140 TI - The IBIS project: data collection in London. Improved Monitoring for Brain Dysfunction during Intensive Care and Surgery. AB - The primary aim of the Improved Monitoring for Brain Dysfunction during Intensive Care and Surgery (IBIS) project was to create a unique and comprehensively annotated data library (DL) of multiple physiological, including neurophysiological, signals. Data collection was undertaken in Kuopio, Finland and London, UK, and comparable protocols were used at all the sites. In London, 43 patients were recruited at the Royal Brompton Hospital, followed by nine at St. Bartholomew's Hospital, all of whom underwent cardiac or combined cardiac and carotid artery surgery. Thirty-seven patients underwent a single operation, while 15 underwent two procedures. The protocols and equipment used, problems specific to the electrically hostile environment and preliminary results are described, including those of clinical interest. The DL is being used for the development of clinically applicable neurophysiological monitoring tools. PMID- 11064141 TI - Technical description of the IBIS data library. Improved Monitoring for Brain Dysfunction in Intensive Care and Surgery. AB - The IBIS Data Library (DL) is an annotated data library that contains practically all the monitored data and other clinical information from critically ill patients during surgery and in intensive care. The data have been collected at three sites: the intensive care unit of the Kuopio University Hospital, Finland; Royal Brompton Hospital, London, UK; and St. Bartholomew's Hospital, London, UK. The purpose of the DL is to form the basis for development of biosignal interpretation methods in the Improved Monitoring for Brain Dysfunction in Intensive Care and Surgery project in the European Union (EU) BIOMED2 programme (BMH4-97-2570). The DL contains continuous electroencephalography signals, multimodal evoked potential recordings and diagnostic electrocardiography recorded during intensive care and surgery. In addition, signal types similar to those recorded during an earlier project, the EU-BIOMED1 project IMPROVE, are stored in the DL. In addition, trend data from patient monitors, laboratory data, annotations, nursing actions, and medications recorded and stored by a Patient Data Management System (PDMS) during routine care are included. The data obtained routinely are complemented by special annotations made by a physician who observes the patient during the data collection session. Annotations include, for example, assessment of the awareness of the patient and specific events during surgery not recorded routinely by the PDMS. Inclusion of information about the care plan and the aims of the care make the contents of the DL complete. The present paper describes the technical set-up used for recording of the DL and the contents of the DL. The paper also includes an appendix defining a new data format, the extended evoked potentials format, used for storage of sweep data in the DL. PMID- 11064142 TI - Exploring the IBIS data library contents: tools for data visualisation, (pre-) processing and screening. Improved Monitoring for Brain Dysfunction in Intensive Care and Surgery. AB - During the IBIS project a high-quality data library of continuous and intermittent physiological signals and variables from patients during intensive care and surgery has been collected. To facilitate exploration of the full content of this data library a data browser was developed, which offers a flexible graphical display of the collection of multivariate data. To supplement the functionality of the display of the 'raw' data, a set of screening and pre processing tools has been developed. A separate trend analysis tool offers a convenient overview of an entire recording focusing on the slow changes in the general state of the patient and the interaction between different physiological subsystems seen from a long-term perspective. A frequency analysis tool for processing the electroencephalography (EEG) signals has been integrated in the data browser to facilitate a quick screening of the cerebral function. The data library is the foundation of the development and validation of biosignal interpretation methods. This process can potentially be more productive using the described tool for algorithm prototyping based on a graphical network specifying the interaction between data processing primitives. PMID- 11064143 TI - Detection of artifacts in monitored trends in intensive care. AB - In intensive care, decision-making is often based on trend analysis of physiological parameters. Artifact detection is a pre-requisite for interpretation of trends both for clinical and research purposes. In this study, we developed and tested three methods of artifact detection in physiological data (systolic, mean and diastolic artery and pulmonary artery pressures, central venous pressure, and peripheral temperature) using pre-filtered physiological signals (2-min median filtering) from 41 patients after cardiac surgery. These methods were: (1) the Rosner statistic; (2) slope detection with rules; and (3) comparison with a running median (median detection). After tuning the methods using data from 20 randomly chosen patients, the methods were tested using the data from the remaining patients. The results were compared with those obtained by manual identification of artifacts by three senior intensive care unit physicians. Out of an average of 22,480 data points for each variable, the three observers labelled 0.98% (220 data points) as artifacts. The inter-observer agreement was good. The average (range) sensitivity for artifact detection in all variables in the test database was 66% (33-92%) for the Rosner statistic, 64% (24 98%) for slope detection and 72% (41-98%) for median detection. All methods had a high specificity (> or = 94%). Slope detection had the highest mean positive prediction rate (53%; 21-85%). When the performance was measured by the cost function, slope detection and running median performed equally well and were superior to Rosner statistics for systemic arterial and central venous pressure and peripheral temperature. None of the methods produced acceptable results for pulmonary artery pressures. We conclude that median filtering of physiological variables is effective in removing artifacts. In post-operative cardiac surgery patients, the remaining artifacts are difficult to detect among physiological and pathophysiological changes. This makes large databases for tuning artifact algorithms mandatory. Despite these limitations, the performance of running median and slope detection were good in selected physiological variables. PMID- 11064144 TI - Quantification of haemodynamic response to auditory stimulus in intensive care. AB - Measuring effects of sensory stimuli on haemodynamics could provide information about the interplay between central and autonomous nervous system (ANS). However, ANS response to sensory stimulus has received little attention. In this paper we present a signal processing scheme to extract the responses of heart rate and systemic arterial pressure on auditory stimulus in intensive care patients (N=5). In short, the effect of mechanical ventilation is rejected by optimal linear modelling. Other disturbances are attenuated by filtering and efficient rejection of outlying sweeps of data. The results show identifiable responses on three out of five cases. The response characteristics may be explained by synchronisation of spontaneous variability in systemic arterial pressure to auditory stimulus. PMID- 11064145 TI - Single sweep analysis of event related auditory potentials for the monitoring of sedation in cardiac surgery patients. AB - Event-related potentials (ERPs) from the auditory system were investigated in 28 post-operative cardiac patients in order to assess their relevance in the monitoring of patient sedation level. Midazolam (17 patients) and propofol (11 patients) were the sedative agents used. The auditory ERP components of N100 (HAB100) and mismatch negativity (MMN) were considered. A single sweep method based on the AutoRegressive with eXogenous input (ARX) model, which is able to enhance the evoked responses to each single stimulus, was used to process each sweep and to compute traditional parameters on a sweep-by-sweep basis. Differences in the measured parameters were related to variations in the patient sedation levels classified through Ramsay score. Significant differences (P<0.05) in both MMN and HAB100 parameters were found between light sedation (LS) and deep sedation (DS) levels. PMID- 11064146 TI - The IBIS system architecture. Improved Monitoring for Brain Dysfunction in Intensive Care and Surgery. AB - An efficient method of interfacing between patient related data and the signal processing methods used to interpret these data is a critical component for a successful patient monitor, particularly for use in high dependency environments. This paper describes the approach adopted in the IBIS project. The key elements of the system are first introduced. A multi-layer structure is then developed in which the outputs from one decision layer are used to enhance the information available for subsequent layers in the system. The resulting system is modular, robust, flexible and easy to modify. PMID- 11064147 TI - Activation of the Jak/Stat signal transduction pathway in GH-treated rat osteoblast-like cells in culture. AB - In this study, activation of the Jak/Stat signaling pathway was followed upon growth hormone (GH) stimulation, using the rat osteosarcoma cell-line UMR-106.01 that expresses high affinity GH receptors. The results show a GH-induced and sustained phosphorylation of Jak2 and Stat5 on tyrosine residues. The tyrosine phosphorylation status of Jak2 was increased in a dose-dependent manner. In contrast to Jak2, tyrosine phosphorylation of Stat5, also elicited at 42 ng/ml GH, remained unchanged when GH concentration was raised up to 4200 ng/ml. DNA binding activity of Stat5 was also observed in response to GH. However, GH was unable to cause transactivation of reporter gene constructs harboring Stat5 binding sites (the GHREII from the rat spi 2.1 gene promoter, and the LHRE from the rat beta-casein gene promoter), except in cells transiently transfected with either Stat5 cDNAs or the rat GHR cDNA. Altogether the results suggest that UMR 106.01 cells exhibit original features of the GH-dependent Jak/Stat signaling pathway. PMID- 11064148 TI - Leptin activates Stat3, Stat1 and AP-1 in mouse adipose tissue. AB - Intraperitoneal leptin administration to wild-type and ob/ob mice caused a prompt activation of Stat1 and Stat3, the former to a lesser extent, in epididymal adipose tissue. Immunoblot experiments showed that tyrosine phosphorylation of Stat3 increased in total cellular extracts and that the phosphorylated protein translocated into the nucleus upon leptin treatment. Tyrosine phosphorylation and nuclear translocation of Stat1 were evident only in ob/ob mice. Gel shift and supershift analyses showed that leptin activated sis-inducible element (SIE) binding activity of adipose nuclear extracts, with Stat3 homodimer as the predominant complex. Stat1/3 heterodimers and Stat1 homodimers take part as well in the response in wild-type and ob/ob mice, although to a lesser degree. AP-1 binding activity was also induced in adipose tissue by in vivo leptin treatment with a time course that suggests a post-transcriptional inductive mechanism. This effect was greater in the ob/ob than in wild-type mice. Our data indicate that leptin operates in vivo directly on adipose tissue by triggering responses that modulate gene expression. PMID- 11064149 TI - Apparent coactivation due to interference of expression constructs with nuclear receptor expression. AB - Transient cotransfection in COS-7 cells, a standard approach to demonstrate coactivation, was used to study the coactivation properties of NuRIP183, a new nuclear receptor interacting protein of 183 kDa, isolated by a yeast two-hybrid screening. Transfection with a NuRIP183 expression construct strongly increased the ligand-dependent response of reporter constructs for several nuclear receptors when compared to transfection with the empty expression vector. A more detailed study, however, revealed major changes in the expression level of the nuclear receptors in cotransfection experiments, indicating that the observed changes in receptor activity were not due to coactivation but to differences in receptor concentration caused by interference from the cotransfected expression constructs with the expression of the receptor. Such interference, which is inversely related to the length of the insert, was observed, not only in COS-7 cells but also in CV-1 and MCF-7 cells, using different transfection techniques (FuGENE-6 and calcium phosphate) and different expression vectors (pSG5, pcDNA1.1 and pIRESneo). These data cast some doubt on coactivation of nuclear receptors based on similar cotransfection experiments without measurement of receptor concentration. Moreover, it is recommended to limit the amounts of (co)transfected expression plasmid and to avoid the use of empty expression plasmid as a control. Finally, one should be aware of similar misleading results in other experimental set-ups based on cotransfection. PMID- 11064150 TI - Cyclic AMP-dependent and independent stimulations of ovarian steroidogenesis by brain factors in the blowfly, Phormia regina. AB - The involvement of cyclic-AMP (cAMP) as a potential second messenger in the neurohormonal control of ovarian steroidogenesis was investigated in the adult female blowfly Phormia regina. Individual measurements of ovarian cAMP concentrations and of ovarian biosynthesis of ecdysteroids, stimulated after a protein meal, demonstrated that steroidogenesis is preceded by a peak of cAMP in the ovaries. In vitro, ovarian steroidogenesis was stimulated by cell-permeable analogues of cAMP and by forskolin. Crude brain extracts were also able to elicit a rise of cAMP in the ovaries in vitro and the secretion of ecdysteroids into the medium: such extracts were more active before than after the protein meal, suggesting a rapid release of neuroendocrine material after feeding. Extracts were then prepared from the dorso-medial part of the brain, containing the neurosecretory cells of the pars intercerebralis (PI): these extracts were again found to stimulate the ovarian ecdysteroid secretion, but surprisingly, they failed to trigger a rise of cAMP in the ovaries in vitro. However, extracts from the rest of the cephalic nervous mass, deprived of PI, were also steroidogenic and they increased ovarian cAMP. Experiments with Rp-cAMPS, a cAMP antagonist, were not able to prevent the ecdysteroid stimulation by PI extracts, but did so partly for the extracts deprived of PI. This study thus indicates that at least two different cephalic factors are able to stimulate ovarian steroidogenesis in the blowfly, one elaborated by PI and acting via a cAMP-independent mechanism, and the other elaborated outside PI and using cAMP as a second messenger. PMID- 11064151 TI - Phex cDNA cloning from rat bone and studies on phex mRNA expression: tissue specificity, age-dependency, and regulation by insulin-like growth factor (IGF) I in vivo. AB - Phosphate regulating gene with homology to endopeptidases on the X chromosome (Phex) inactivating mutations cause X-linked hypophosphatemia (XLH). The disorder is characterized by decreased renal phosphate (Pi) reabsorption in both humans and mice, in the latter shown to be due to a reduction in mRNA and protein of type II sodium-dependent phosphate cotransporter (NadPi-II). To gain insight into the physiological role of Phex, we cloned the rat cDNA and examined tissue specific and age-dependent mRNA expression. The rat full-length cDNA (2247 nucleotides) shares 96 and 90% identity with the mouse and human cDNA, respectively. We found 6.6 kb Phex transcripts in calvarial bone and lungs, and a weaker signal in liver of newborn rats. In adult animals, Phex mRNA signals were weaker in bone and lungs and absent in liver. Phex mRNA expression in bones and NadPi-I and -II cotransporter mRNA expression in kidney were also determined in hypophysectomized rats. These rats, which lack GH and IGF I, stop growing and exhibit decreased serum Pi levels. Treatment during 6 days with IGF I stimulated growth and increased serum Pi. Phex and NadPi-II cotransporter mRNA levels were higher in IGF I than in vehicle-treated animals, while mRNA expression of NadPi I, 1alpha-hydroxylase and 24-hydroxylase and serum levels of calcitriol remained unaffected. Age-dependency of Phex expression suggests a role for Phex in Pi retention during growth. Moreover, our findings indicate that an increase in Phex expression in bones under the influence of IGF I may contribute to increased serum Pi by enhancing renal phosphate reabsorption. Because IGF I treatment increased NadPi-II mRNA expression and serum Pi, IGF I appears to act at least partially at pretranslational levels to increase NadPi-II mediated renal Pi retention in growing rats. PMID- 11064152 TI - Establishment and characterization of steroidogenic granulosa cells expressing beta(2)-adrenergic receptor: regulation of adrenodoxin and steroidogenic acute regulatory protein by adrenergic agents. AB - Primary granulosa cells obtained from PMSG primed immature rats were triple transfected with SV40 DNA, Ha-ras oncogene and an expression vector containing human beta(2)-adrenergic receptors resulting in granulosa cell lines constitutively expressing the beta(2)-adrenergic receptors. Isoproterenol, a potent adrenergic agent, stimulated both cAMP accumulation and progesterone production in these cells in a dose dependent manner. Responsiveness of these cells was specific only to isoproterenol, while hCG (2.4 nM) and hFSH (2.4 nM) had no effect on steroid production. ED(50) for stimulation of cAMP and progesterone in these cells by isoproterenol was 2x10(-6) M and 7x10(-6) M, respectively. Forskolin also showed a dose dependent stimulation of cAMP and progesterone with ED(50) of 1.5 and 0.35 microg/ml, respectively. Epinephrine at a dose of 10(-5) M elicited maximum response to produce cAMP and progesterone. Isoproterenol induced accumulation of cAMP and progesterone in these cells were inhibited by beta(2)-adrenergic blocker, propranolol with an ED(50) of 6x10(-8) and 7x10(-9) M, respectively, whereas the beta(1)-adrenergic blocker, metoprolol was effective only at a very high concentration (ED(50)>10(-4) and 1.9x10(-5) M for inhibiting isoproterenol induced cAMP and progesterone production, respectively). Induction of steroidogenesis by isoproterenol or forskolin involved de novo synthesis of the cytochrome P450 side chain cleavage (SCC) enzyme complex, as assessed by indirect immunofluorescence staining for adrenodoxin. Western analysis indicate that expression of adrenodoxin is upregulated by forskolin, isoproterenol and adrenalin by 7.8-, 6.9- and 10.8 fold, respectively. The presence of StAR protein was identified by Western blotting. StAR expression was elevated by 8.3-, 2.5- and 4.7-fold upon stimulation with forskolin, isoproterenol and adrenalin, respectively. Thus, this cell line could serve as a good model system to study catecholamine mediated regulation of growth and differentiation of granulosa cells and the role of oncogenes in this process. PMID- 11064153 TI - Characterization of a pituitary GnRH-receptor from a perciform fish, Morone saxatilis: functional expression in a fish cell line. AB - Gonadotropin-releasing hormones (GnRHs) bind to the specific receptor present on the gonadotrophs to activate the synthesis and release of gonadotropins (follicle stimulating hormone or FSH and luteinizing hormone or LH), which in turn control gonadal maturation, gametogenesis and gamete release. Perciform species have three endogenous GnRHs. The main objective of this study was to characterize the gonadotropin-releasing hormone receptor (GnRH-R) present in the pituitary of a perciform species, striped bass (Morone saxatilis) and demonstrate how it interacts with its potential ligand. In this study, a cDNA for GnRH-R from the pituitaries of striped bass was cloned. The cloned cDNA has an open reading frame (ORF) that codes for a 419 amino acids peptide. Like other G-protein coupled receptors including the non-mammalian GnRH-Rs, the peptide has seven putative transmembrane domains and a C-terminal tail. Comparative analysis of the amino acid sequence of striped bass (stb) GnRH-R shows 38-87% similarity with the known GnRH-Rs. A Northern blot analysis revealed a single GnRH-R transcript in the pituitary; however, its expression in various extrapituitary tissues was demonstrated by a reverse-transcription-PCR (RT-PCR). Functionally, upon induction by endogenous forms of GnRHs (seabream, chicken II and salmon GnRHs) and a mammalian GnRH-agonist, the recombinant stbGnRH-R mediated a reporter gene (luciferase) activity in a fish cell line (CHSE-214). A real-time relative quantitation method established that significantly higher (P<0.05) levels of stbGnRH-R mRNA were present in the pituitaries of striped bass with advanced stages of ovarian development, compared to the pituitaries of fish with less developed ovaries. PMID- 11064154 TI - Biochemical and genetic characterization of the porcine Prophet of Pit-1 pituitary transcription factor. AB - Prophet of Pit-1 (Prop-1) is a paired class homeodomain transcription factor that is specifically expressed in the pituitary gland. Mutations in the Prop-1 gene cause compound pituitary diseases in mouse models and human patients. We have cloned and analyzed the porcine ortholog of Prop-1. Analysis of cDNAs revealed that the porcine Prop-1 sequence is similar to the mouse and human proteins within the homeodomain and carboxyl terminus, but the amino terminus is poorly conserved. The Prop-1 gene consists of three exons and two introns and spans 3.8 kilobases of genomic DNA. In addition, we mapped Prop-1 to the q arm of pig chromosome two. During development, Prop-1 is expressed at the time of pituitary organogenesis. In the adult, expression was observed at low levels only in the pituitary gland. The porcine Prop-1 protein displays similar biochemical, DNA binding, and transcriptional activities to human PROP-1. We conclude that, although the structural divergence between the porcine and human PROP-1 molecules may indicate some distinct functions, the porcine Prop-1 gene encodes a pituitary transcription factor with similar overall activities to the human ortholog. PMID- 11064155 TI - An androgen response element mediates LNCaP cell dependent androgen induction of the hK2 gene. AB - Human glandular kallikrein (hK2) is an androgen regulated protein primarily expressed in the prostate and recently identified as a novel prostate cancer marker. A 5 kb 5' flanking region of the hK2 gene was isolated and sequenced to characterize the regulatory mechanisms for the expression of hK2 in the androgen responsive prostate cell line, LNCaP. Using gene transfer, gel shift, and mutagenesis assays we have identified an ARE in the 5' far upstream promoter region of the hK2 gene that is crucial for its regulation in LNCaP cells. This study further demonstrated that the hK2 upstream ARE plays a predominant role in androgenic response. More interestingly, previously identified AREs in the prostate specific antigen promoter and the hK2 proximal promoter exert little activity in LNCaP cells. This study for the first time identifies a unique ARE that alone mediates the function of the androgen receptor in LNCaP cells in a cell dependent manner. This study also examines the activity of this ARE with 1alpha, 25 dihydroxy-vitamin D3 on the expression of the hK2 gene in LNCaP cells. PMID- 11064156 TI - The pigmentotropic hormone [His(7)]-corazonin, absent in a Locusta migratoria albino strain, occurs in an albino strain of Schistocerca gregaria. AB - [His(7)]-corazonin has recently been identified in the corpora cardiaca (CC) of two locust species, the migratory locust, Locusta migratoria and the desert locust, Schistocerca gregaria, as the dark colour inducing neurohormone. Here, we investigate whether [His(7)]-corazonin occurs in the brain-CC axis of a Schistocerca albino strain. From data obtained by immunocytochemistry, injection experiments, chromatographic and mass spectrometric analysis of brain and CC tissues, it could be concluded that an albino strain of S. gregaria from Denmark contains authentic [His(7)]-corazonin. This was unequivocally demonstrated by sequencing the [His(7)]-corazonin-immunoreactive factor in albino Schistocerca brain-CC extracts with ESI-Qq-oa-TOF mass spectrometry. Albinism in this strain is hence not caused by the deficiency of authentic [His(7)]-corazonin in the brain-CC axis, nor by defects in release. Conversely to L. migratoria albinos, injection of [His(7)]-corazonin failed to induce dark pigmentation in Schistocerca albinos. Therefore, albinism in the investigated Schistocerca strain is likely to be situated at the level of the receptor, signal transduction mechanisms or of pigment biosynthesis. PMID- 11064157 TI - Activin maintains the condensed type of mitochondria in germ cells. AB - Development of germ cells during spermatogenesis is characterized by a complex series of differentiation events finally leading to the production of spermatozoa. Beside the main hormonal regulators, paracrine interactions are thought to play a major role in this process. Mitochondria in germ cells pass through unique alterations ranging from the 'typical' cristae-rich mitochondria found in spermatogonia to the 'condensed' form in pachytene spermatocytes. This study provides further support that paracrine factors produced by Sertoli cells, most likely activin A, are involved in germ cell differentiation as monitored by the maintenance of the physiological 'condensed' mitochondrial phenotype in primary spermatocytes. Culture of primary spermatocytes in Sertoli cell conditioned medium (SCCM) for 18 h resulted in the maintenance of a high percentage of 'condensed-type' mitochondria in comparison to cells cultured in Dulbecco's minimum essential medium (DMEM). Activin A, a product of Sertoli cells, showed at subnanogram concentrations a similar ability to SCCM to maintain a high percentage of spermatocyte mitochondria in the 'condensed' state, while inhibin had no effect. The addition of an antiserum specific for activin A resulted in a neutralization of the effect caused by activin A or SCCM. This strongly suggested that the active substance in SCCM was activin A. Taken together these data indicate that activin A is the first Sertoli cell product that has been identified to influence differentiation of male meiotic germ cells. PMID- 11064158 TI - Conservation of steroidogenic acute regulatory (StAR) protein structure and expression in vertebrates. AB - Complementary DNAs for the open reading frames of the chicken, Xenopus and zebrafish StAR homologs were cloned along with a partial cDNA of the zebrafish homolog to MLN64, a StAR-related protein. A comparison of the amino acid sequences of piscine, amphibian, avian and mammalian StARs, indicates strong conservation of the protein across divergent vertebrate groups. On Northern blots probed with species specific StAR cDNAs, expression of StAR transcripts was observed in the ovary and adrenal of chicken, and the ovary, testis, kidney and head of zebrafish. The expression of StAR mRNA in various compartments of the hen ovary was consistent with the results of past studies on steroidogenesis; expression was first observed in follicles selected into the preovulatory hierarchy and was greatest in the largest preovulatory follicle. The expression of StAR mRNA was also consistent with aromatase expression in zebrafish ovaries. The conserved deduced protein sequence and expression pattern of StAR transcripts in chicken and zebrafish tissues, strongly suggest that StAR is also involved in the regulation of steroidogenesis in nonmammalian vertebrates. PMID- 11064159 TI - Effect of thyroid hormones on mitochondrial oxygen free radical production and DNA oxidative damage in the rat heart. AB - Mitochondria seem to be involved in oxygen radical damage and aging. However, the possible relationships between oxygen consumption and oxygen radical production by functional mitochondria, and oxidative DNA damage, have not been studied previously. In order to analyze these relationships, male Wistar rats of 12 weeks of age were rendered hyper- and hypothyroid by chronic T(3) and 6-n-propyl-2 thiouracil treatments, respectively. Hypothyroidism decreased heart mitochondrial H(2)O(2) production in States 4 (to 51% of controls; P<0.05) and 3 (to 21% of controls; P<0.05). In agreement with this, 8-oxo-7,8-dihydro-2'-deoxyguanosine (8 oxodG) decreased in the heart genomic DNA of hypothyroid animals to 40% of controls (P<0.001). Studies with respiratory inhibitors showed that the decrease in oxygen radical generation observed in hypothyroidism occurred at Complex III (mainly) and at Complex I; that decrease was due to the presence of a lower free radical leak in the respiratory chain (P<0.05). Hyperthyroidism did not significantly change heart mitochondrial H(2)O(2) production since the increase in State 4 oxygen consumption in comparison with control and hypothyroid animals (P<0.05) was compensated by a decrease in the free radical leak in relation to control animals (P<0.05). In agreement with this, heart 8-oxodG was not changed in hyperthyroid animals. The lack of increase in H(2)O(2) production per unit of mitochondrial protein will protect mitochondria themselves against self-inflicted damage during hyperthyroidism. PMID- 11064160 TI - Complementary DNA structure and genomic organization of Drosophila menin. AB - Menin is a protein product of a tumor supressor gene MEN1, mutations of which are responsible for multiple endocrine neoplasia type 1, an autosomal dominant familial cancer syndrome. We determined the nucleotide sequence of the Drosophila menin cDNA using RT-PCR and RACE, and confirmed it by direct sequencing of genomic DNA. Gene expression of Drosophila menin was detected by Northern blot analysis in adult and embryo as two types of transcripts, one identical in size to the cDNA, and the other larger but detected only in embryo. The Drosophila menin gene was composed of five exons in which the protein was encoded in exon 2 through 5, and spanned approximately 6.3 kb. The deduced amino acid (AA) sequence of Drosophila menin consisted of 751 AAs with a calculated molecular mass of 81.7 kDa, and showed 44-47% identity to human, rat, mouse and zebrafish menin over the entire length. Among the AA residue substitutions that have been reported as disease-associated missense mutations and single AA deletions, 53 out of 71 were completely conserved in Drosophila. The presence of menin ortholog in insect indicates that menin is an evolutionally conserved protein with a fundamental role in biological processes. PMID- 11064178 TI - Dorothy Hatsukami and the sponsorship of Drug and Alcohol Dependence by the College on Problems of Drug Dependence. PMID- 11064179 TI - Developmental sources of variation in liability to adolescent substance use disorders. AB - This review provides a synthesis of the literature on the complex sequence of maturational, psychosocial, and neuroadaptive processes that lead to substance use disorders (SUD) in adolescence. A brief overview introduces the concepts of liability to SUD and epigenesis. A theory is presented explaining how affective, cognitive, and behavioral dysregulation in late childhood is exacerbated during early and middle adolescence by family and peer factors, as well as puberty, leading to substance use. Continued exacerbation of the three components of dysregulation by drug and non-drug stressors during late adolescence is posited to result in neuroadaptations that increase the likelihood of developing SUD, particularly in high-risk individuals. Implications for etiologic research as well as clinical and preventive interventions are discussed. PMID- 11064180 TI - A cocaine analog, 2beta-propanoyl-3beta-(4-tolyl)-tropane (PTT), reduces tyrosine hydroxylase in the mesolimbic dopamine pathway. AB - Tyrosine hydroxylase (TH) is the rate-limiting enzyme in catecholamine biosynthesis. Previously published results have established that chronic cocaine administration (30-45 mg/kg per day, 10-14 days) resulted in an upregulation of TH gene expression in dopaminergic pathways of rats. The present studies tested the effects of a tropane analog, PTT (2beta-propanoyl-3beta-(4-tolyl)-tropane), on TH expression. This drug has similar actions to cocaine, but possesses markedly different pharmacokinetics (20 times more potent at binding the dopamine transporter, markedly increased metabolic stability, and 10-20 times more potent in behavioral measures). Moreover, PTT demonstrates an increased selectivity for the dopamine (DA) and norepinephrine (NE) transporters compared with cocaine. In direct contrast to the previously reported effects of cocaine, 10 days of PTT administration (3.0 mg/kg per day, i.p.) produced a uniform downregulation of TH protein and activity gene expression. TH activity and immunoreactive protein where decreased by 54 and 69%, respectively in the nucleus accumbens. Within the ventral tegmental area, TH activity and protein were decreased by 33 and 19%, respectively. The underlying mechanisms for these fundamental differences are unclear, but likely reflect varying and selective affinities and lengths of occupancy at biogenic amine transporters. PMID- 11064181 TI - Sociopathy, gender, and treatment outcome among outpatient substance abusers. AB - Antisocial personality disorder (ASPD) may predict poor prognosis but gender/sociopathy relationships to prognosis remain unclear. This study investigated the effects of ASPD upon psychiatric and substance-related outcomes among 235 addiction treatment center outpatients. Prevalence rates for ASPD were similar for males (16%) and females (22%). At baseline, women and ASPD patients displayed greater substance-related and psychiatric severity. At 6-month follow up, ASPD patients had greater severity on both measures than did patients without ASPD, but women now had equivalent psychiatric severity to men. After controlling for initial severity, ASPD was related to worse substance-related outcomes, but not to worse psychiatric outcomes. PMID- 11064182 TI - Cocaine, HIV, and their cardiovascular effects: is there a role for ACE-inhibitor therapy? AB - Cocaine abuse and HIV disease each have potentially adverse effects upon the heart and cardiovascular system which may be exacerbated when these risk factors are combined. The development of a safe and effective agent to treat both cocaine addiction and its cardiovascular sequelae, that is well-tolerated by HIV patients, would thus be of considerable clinical utility. In this article we discuss the rationale for the investigation of angiotensin converting enzyme (ACE) inhibitors, commonly used to treat hypertension, for treatment in cocaine abusing populations, based on their potential to reduce cocaine use by modulating levels of dopamine and corticotropin releasing factor in the brain, and on their ability to reverse cardiovascular and platelet abnormalities. We present preliminary findings from echocardiographic and platelet activation studies in 16 HIV-positive, cocaine abusing patients, as well as tolerability and efficacy studies of the ACE-inhibitor, fosinopril, for the treatment of cocaine abuse in both HIV-positive (n=6) and HIV-negative (n=5) methadone-maintained cocaine abusers. Findings suggest that HIV-positive cocaine-abusing patients possess abnormalities of diastolic heart function and platelet activation that are potentially reversible with ACE-inhibitor therapy. Findings also suggest that fosinopril is well-tolerated regardless of HIV serostatus, does not appear to cause hypotension, and may possess effectiveness for reducing cocaine use. We conclude that ACE-inhibitor therapy may offer a new pharmacologic approach to the treatment of cocaine abuse and its complications, and that controlled research of this class of agents may be promising. PMID- 11064183 TI - Plasma concentrations of the enantiomers of methadone and therapeutic response in methadone maintenance treatment. AB - Methadone is a 50:50 mixture of two enantiomers and (R)-methadone accounts for the majority of its opioid effect. The aim of this study was to determine whether a blood concentration of (R)-methadone can be associated with therapeutic response in addict patients in methadone maintenance treatment. Trough plasma concentrations of (R)-, (S)- and (R,S)-methadone were measured in 180 patients in maintenance treatment. Therapeutic response was defined by the absence of illicit opiate or cocaine in urine samples collected during a 2-month period prior to blood sampling. A large interindividual variability of (R)-methadone concentration-to-dose-to-weight ratios was found (mean, S.D., median, range: 112, 54, 100, 19-316 ng x kg/ml x mg). With regard to the consumption of illicit opiate (but not of cocaine), a therapeutic response was associated with (R)- (at 250 ng/ml) and (R,S)-methadone (at 400 ng/ml) but not with (S)-methadone concentrations. A higher specificity was calculated for (R)- than for (R,S) methadone, as the number of non-responders above this threshold divided by the total number of non-responders was higher for (R,S)-methadone (19%) than for (R) methadone (7%). The results support the use of therapeutic drug monitoring of (R) methadone in cases of continued intake of illicit opiates. Due to the variability of methadone concentration-to-dose-to-weight ratios, theoretical doses of racemic methadone could be as small as 55 mg/day and as large as 921 mg/day to produce a plasma (R)-methadone concentration of 250 ng/ml in a 70-kg patient. This demonstrates the importance of individualizing methadone treatment. PMID- 11064184 TI - Zaleplon and triazolam: drug discrimination, plasma levels, and self administration in baboons. AB - Zaleplon is a chemically novel hypnotic that preferentially binds alpha(1) subunit containing subtypes of the alphabetagamma configuration of the gamma aminobutyric acid (GABA)(A) receptor. Zaleplon and the non-subtype-selective hypnotic triazolam occasioned 100% drug-appropriate responding in baboons trained to discriminate lorazepam or pentobarbital from vehicle. Flumazenil shifted the zaleplon generalization gradient at least five-fold to the right. A plasma elimination half-life of 6-8 h for oral 10 mg/kg zaleplon and 0.32 mg/kg triazolam was paralleled by discriminative control for 7 h. Zaleplon maintained self-injection greater than vehicle, as did comparison doses of the similarly selective hypnotic zolpidem and triazolam. Concurrent food-maintained responding increased during self-injection of all three drugs. Preferential binding at this alpha(1)-containing GABA(A) subtype did not diminish the benzodiazepine (Bzs) like behavioral effects of zaleplon. PMID- 11064185 TI - Zaleplon and triazolam physical dependence assessed across increasing doses under a once-daily dosing regimen in baboons. AB - The ability of the GABA(A)-receptor-subtype-selective hypnotic zaleplon to produce physical dependence was compared to the nonselective benzodiazepine triazolam. Progressively increasing doses of zaleplon and triazolam were given to baboons by intragastric infusion once each day, with doses increasing every 17 days. Next, the highest dose was given for 10-34 additional days by continuous infusion. Both drugs produced increases in food-maintained lever pressing, ataxia, and time to complete a fine motor task. Plasma levels increased dose dependently; drug was detectable 24 h after higher doses. Flumazenil produced a mild or intermediate precipitated-withdrawal syndrome on day 14 of all dosing conditions. When drug delivery ended after 85-100 days, a benzodiazepine-type withdrawal syndrome occurred. Physical dependence potential of zaleplon and triazolam appear similar. PMID- 11064186 TI - Buprenorphine and naloxone co-administration in opiate-dependent patients stabilized on sublingual buprenorphine. AB - Buprenorphine and naloxone sublingual (s.l.) dose formulations may decrease parenteral buprenorphine abuse. We evaluated pharmacologic interactions between 8 mg s.l. buprenorphine combined with 0, 4, or 8 mg of naloxone in nine opiate dependent volunteers stabilized on 8 mg s.l. buprenorphine for 7 days. Combined naloxone and buprenorphine did not diminish buprenorphine's effects on opiate withdrawal nor alter buprenorphine bioavailability. Opiate addicts stabilized on buprenorphine showed no evidence of precipitated opiate withdrawal after s.l. buprenorphine-naloxone combinations. Buprenorphine and naloxone bioavailability was approximately 40 and 10%, respectively. Intravenous buprenorphine and naloxone produced subjective effects similar to those of s.l. buprenorphine and did not precipitate opiate withdrawal. PMID- 11064187 TI - Genetic and environmental influences on posttraumatic stress disorder, alcohol and drug dependence in twin pairs. AB - We investigated whether and to what degree genetic and environmental contributions overlap among posttraumatic stress disorder (PTSD), alcohol dependence (AD) and drug dependence (DD). Subjects were 3304 monozygotic and dizygotic male-male twin pair members of the Vietnam Era Twin Registry who participated in 1992 telephone administration of the Diagnostic Interview Schedule Version 3 Revised (DIS-3R). Genetic model fitting was performed to estimate the magnitude of genetic and environmental contributions to the lifetime co-occurrence of DSM-III-R PTSD, AD and DD. The liability for PTSD was partially due to a 15.3% genetic contribution common to AD and DD and 20.0% genetic contribution specific to PTSD. Risk for AD was partially due to a 55.7% genetic contribution common to PTSD and DD. Genetic influences common to PTSD and AD accounted for 25.2% of the total risk for DD. Specific family environmental influence accounted for 33.9% of the total variance in risk for DD. Remaining variance for all three disorders was due to unique environmental factors both common and specific to each phenotype. These results suggest that PTSD, AD and DD each have etiologically distinct components and also have significant genetic and unique environmental contributions in common. PMID- 11064188 TI - Bactericidal and inhibitory effects of azole antifungal compounds on Mycobacterium smegmatis. AB - Azole antifungals are central to therapy and act by inhibiting a cytochrome P450, sterol 14-demethylase and blocking normal sterol synthesis. Our recent identification of a mycobacterial sterol biosynthetic pathway led us to probe the efficacy of a range of these compounds against Mycobacterium smegmatis. Several showed equivalent or greater inhibitory effects to those against Candida albicans, and bactericidal activity was demonstrated for four compounds, clotrimazole, econazole, miconazole and tebuconazole. The major drug used clinically, fluconazole, was ineffective. The results are discussed in the light of the world-wide spread of tuberculosis, including drug-resistant forms and the requirement for new drugs. PMID- 11064189 TI - Differential regulation of amoA and amoB gene copies in Nitrosomonas europaea. AB - Nitrosomonas europaea contains two nearly identical copies of the operon, amoCAB, which encodes the ammonia monooxygenase (AMO) enzyme. Cells of N. europaea containing single mutations in either amoA or amoB gene copies were incubated in ammonium both prior to and after exposure to acetylene or light. For each strain, the O(2) consumption rates and amounts of AmoA polypeptide, the active site containing subunit of AMO, produced in each strain were determined. Strains carrying a mutation in either the amoA(2) or amoB(2) genes responded similarly to wild-type cells, but the strains carrying mutations in the amoA(1) or amoB(1) genes responded differently from the wild-type, or from each other. These results suggest that the copies of amoA and amoB are differentially regulated upon exposure to different external stimuli. PMID- 11064190 TI - High catalytic activity of alanine racemase from psychrophilic Bacillus psychrosaccharolyticus at high temperatures in the presence of pyridoxal 5' phosphate. AB - We examined the effect of the pyridoxal 5'-phosphate (PLP) cofactor on the activity and stability of the psychrophilic alanine racemase, having a high catalytic activity at low temperature, from Bacillus psychrosaccharolyticus at high temperatures. The decrease in the enzyme activity at incubation temperatures over 40 degrees C was consistent with the decrease in the amount of bound PLP. Unfolding of the enzyme at temperatures above 40 degrees C was suppressed in the presence of PLP. In the presence of 0.125 mM PLP, the specific activity of the psychrophilic enzyme was higher than that of a thermophilic alanine racemase, having a high catalytic activity at high temperature, from Bacillus stearothermophilus even at 60 degrees C. PMID- 11064191 TI - A transposon carrying the gene mphB for macrolide 2'-phosphotransferase II. AB - The mphB gene for macrolide 2'-phosphotransferase II is located on two plasmids, pTZ3721 and pTZ3723, in Escherichia coli BM2506. We examined translocation of mphB that originated from pTZ3721. The transposable element carrying mphB is 39 kb long and has a Tn21-like transposition module at one end and a Tn1721-like transposition module at other. The structure of the transposition modules of this element resembles that of Tn2610. However, the gene arrangement of the internal region on the transposon carrying mphB was reverse to that of Tn2610. The nucleotide sequences of both terminal regions suggested that the inversion of the DNA fragment occurred between the res sites by resolvase-mediated recombination. PMID- 11064192 TI - Progesterone side-chain cleavage by Bacillus sphaericus. AB - The side-chain of progesterone was cleaved by Bacillus sphaericus to produce two C-19 keto androstene steroids. The structures of these metabolites were androstenedione and 1-dehydroandrostenedione. High concentrations of glucose in the culture medium inhibited conversion of progesterone to these two metabolites. PMID- 11064193 TI - Utilisation of aliphatic compounds by acidophilic heterotrophic bacteria. The potential for bioremediation of acidic wastewaters contaminated with toxic organic compounds and heavy metals. AB - Acidophilic, heterotrophic bacteria isolated from acidic mine effluent metabolised a range of aliphatic compounds. Aliphatic acids, which are normally thought to be toxic to acidophiles, were utilised as substrates for energy and growth by these bacteria. This biodegradative ability, concomitant with their tolerance of heavy metals, has demonstrated the potential for using these organisms for the bioremediation of multiply contaminated acidic wastewaters. PMID- 11064194 TI - Saccharomyces uvarum, a proper species within Saccharomyces sensu stricto. AB - Saccharomyces uvarum is proposed as a proper species within the complex Saccharomyces sensu stricto. Molecular characteristics including the similarity of the restriction profile of the non-transcribed spacer 2 (NTS2) and of the D1/D2 sequences of the rDNA, as well as other genotypic and phenotypic characteristics confirm that this group of strains is highly homogeneous and distinguishable from other species of the Saccharomyces sensu stricto group. PMID- 11064195 TI - alpha-Galactosidase A from Pseudomonas fluorescens subsp. cellulosa: cloning, high level expression and its role in galactomannan hydrolysis. AB - A library of Pseudomonas fluorescens subsp. cellulosa genomic DNA, constructed in lambda ZAPII, was screened for alpha-D-galactosidase activity. The DNA inserts from six galactosidase-positive clones were rescued into plasmids. Restriction digestion and Southern analysis revealed that each of the plasmids contained a common DNA sequence. The sequence of the Pseudomonas DNA in one of the plasmids revealed a single open reading frame (aga27A) of 1215 bp encoding a protein of M(r) 45900, designated alpha-galactosidase 27A (Aga27A). Aga27A exhibited extensive sequence identity with alpha-galactosidases in glycoside hydrolase 27, and appeared to be a single domain protein. The recombinant alpha-galactosidase was expressed at high levels in Escherichia coli and the biophysical properties and substrate specificity of the enzyme were evaluated. The data showed that Aga27A was a mesophilic neutral acting non-specific alpha-galactosidase. Both P. fluorescens subsp. cellulosa mannanase A (ManA) and Aga27A hydrolyse the polymeric substrate, carob galactomannan. Sequential hydrolysis with AgaA followed by ManA, or ManA followed by AgaA enhanced product release. The positive effects of sequential hydrolysis are discussed. PMID- 11064196 TI - Respiratory pathways of Rhodobacter sphaeroides 2.4.1(T): identification and characterization of genes encoding quinol oxidases. AB - Rhodobacter sphaeroides 2.4.1(T) respires aerobically via a branched respiratory chain consisting of both cytochrome c oxidases and quinol oxidases. Here, genes from chromosome II encoding two distinct quinol oxidases have been characterized. The qoxBA genes encode a putative heme-copper quinol oxidase, whereas the qxtAB genes encode a quinol oxidase homologous to the cyanide-insensitive oxidase of Pseudomonas aeruginosa. No phenotype was observed for mutations in either oxidase in the wild-type background. A strain containing a qxtA mutation in a cytochrome bc(1) complex mutant background was unable to grow aerobically. No role was found for the Qox oxidase, nor was a qoxB::lacZ transcriptional fusion expressed under a variety of conditions. These are the first molecular studies to characterize the quinol oxidases of R. sphaeroides 2.4.1(T). PMID- 11064197 TI - Phenotypic analyses of frz and dif double mutants of Myxococcus xanthus. AB - Myxococcus xanthus is a Gram-negative gliding bacterium that aggregates and develops into multicellular fruiting bodies in response to starvation. Two chemosensory systems (frz and dif), both of which are homologous to known chemotaxis proteins, were previously identified through characterization of various developmental mutants. This study aims to examine the interaction between these two systems since both of them are required for fruiting body formation of M. xanthus. Through detailed phenotypic analyses of frz and dif double mutants, we found that both frz and dif are involved in cellular reversal and social motility; however, the frz genes are epistatic in controlling cellular reversal, whereas the dif genes are epistatic in controlling social motility. The study suggests that the integration of these two chemotaxis systems may play a central role in controlling the complicated social behaviors of M. xanthus. PMID- 11064198 TI - The transcriptional activator NtrC controls the expression and activity of glutamine synthetase in Herbaspirillum seropedicae. AB - The role of the Ntr system in Herbaspirillum seropedicae was determined via ntrB and ntrC mutants. Three phenotypes were identified in these mutants: Nif(-), deficiency in growth using nitrate, and low glutamine synthetase (GS) activity. All phenotypes were restored by the plasmid pKRT1 containing the intact glnA, ntrB and ntrC genes of H. seropedicae. The promoter region of glnA was subcloned into a beta-galactosidase fusion vector and the results suggested that NtrC positively regulates the glnA promoter in response to low nitrogen. The H. seropedicae ntrC and ntrB mutant strains showed a deficiency of adenylylation/deadenylylation of GS, indicating that NtrC and NtrB are involved in both transcription and activity control of GS in this organism. PMID- 11064199 TI - Contribution of defined amino acid residues to the immunogenicity of recombinant Escherichia coli heat-stable enterotoxin fusion proteins. AB - We investigated whether the toxicity-associated receptor-binding domain of the non-immunogenic Escherichia coli heat-stable enterotoxin (STh) as a fusion with a carrier protein and the inclusion of an appropriate spacer are critical factors for eliciting antibody responses against the native toxin. The immunological properties of three toxic and one non-toxic fusion proteins, consisting of STh N terminally joined to the C-terminus of the major subunit ClpG of E. coli CS31A fimbriae, were compared. In contrast to the non-toxic hybrid STh with glycine and leucine simultaneously substituted for the receptor-interacting Pro(13) and Ala(14) amino acids, the toxic chimeras responded by producing high serum levels of anti-STh antibodies in immunized animals. On the other hand, only the toxic ClpG-STh construct with the natural peptide 47KSGPESM(53) of Pro-STh as spacer stimulated STh-neutralizing responses against both native toxin and enterotoxigenic live E. coli cells. Altogether, these findings suggest a close relationship between conformational similarity to the native structure of STh and the ability to elicit specific antibody responses against STh. PMID- 11064200 TI - A unique and common restriction fragment pattern of the nucleotide sequences homologous to the genome of vf33, a filamentous bacteriophage, in pandemic strains of Vibrio parahaemolyticus O3:K6 O4:K68, and O1:K untypeable. AB - Vf33 is a filamentous bacteriophage isolated from Vibrio parahaemolyticus. We performed Southern blot hybridization analysis to examine the distribution of Vf33-related genetic elements in the pandemic strains (O3:K6 strains isolated between 1995 and 1997, O4:K68 and O1:K untypeable strains isolated between 1997 and 1999) of V. parahaemolyticus. Nucleotide sequences homologous to the Vf33 DNA were detected in all 57 test strains including pandemic and non-pandemic strains. However, the profiles of hybridization, including the restriction fragment length polymorphism, with nine Vf33-derived DNA probes exhibited by the pandemic strains were identical and were different from those by the non-pandemic strains. The results support the hypothesis that the pandemic strains are clonal, and suggest a possibility that they have acquired (a) new gene(s) via a Vf33-like filamentous phage. PMID- 11064201 TI - Suppression of temperature-sensitive sporulation mutation in the Bacillus subtilis sigA gene by rpoB mutation. AB - We isolated a temperature-sensitive sporulation defective mutant of the sigA gene, encoding a major sigma factor, sigma(A) protein, in Bacillus subtilis, and designated it as sigA21. The sigA21 mutation caused a single-amino acid substitution, E314K, in region 4 of the sigma(A) protein. In this mutant, expression of the spoIIG gene, whose transcription depends on both sigma(A) and the phosphorylated Spo0A protein, Spo0A approximately P, a major transcription factor during early stages of sporulation, was greatly reduced at 43 degrees C. To obtain further information on the mechanism of sigma(A) function during the early spore development, we isolated a spontaneous sporulation-proficient suppressor mutant at 43 degrees C. This extragenic suppressor mutation was mapped within the rpoB gene, encoding the beta subunit of RNA polymerase, and was found to have a single-amino acid substitution, A863G. In this mutant, the expression of the spoIIG is partially restored at 43 degrees C. PMID- 11064202 TI - Cytochemical evaluation of localization and secretion of a heterologous enzyme displayed on yeast cell surface. AB - A starch-utilizing Saccharomyces cerevisiae strain was constructed by cell surface engineering. Distribution of the heterologous glucoamylase-alpha agglutinin fusion protein on the yeast cell was analyzed by indirect fluorescence microscopy using an anti-glucoamylase antibody. Most of the intense fluorescence was first localized in the small bud, then observed on the entire cell wall of the daughter and mother cells. Fluorescence also accumulated at the neck region. These observations suggest that the display of the heterologous protein on the cell surface is carried with other cell wall components to the areas in which the cell wall is newly synthesized; the distribution is controlled by the cell cycle. Then, the heterologous protein-encoding gene was expressed in a sec1 mutant, in which secretory vesicles accumulate under restrictive temperature, and the produced protein was detected by immunoelectron microscopy. Most of the gold particles that reacted with the fusion protein were not localized in vesicles but in expanding endoplasmic reticulum. This phenomenon may be due to overproduction of the heterologous protein which was designed to be displayed on the cell wall. Artificial production of heterologous protein may have caused a relative shortage of glycosyl phosphatidylinositol anchors. PMID- 11064203 TI - Characterisation of a Pasteurella multocida esterase gene which confers a hemolytic phenotype in Escherichia coli under anaerobic conditions. AB - Investigation of the hemolytic phenotype under anaerobic growth conditions of an avian Pasteurella multocida strain, PBA100, resulted in the identification and characterisation of a gene encoding an esterase enzyme, mesA, that conferred a hemolytic phenotype in Escherichia coli under anaerobic conditions. MesA appeared to be expressed and functional under anaerobic and aerobic conditions in both E. coli and P. multocida. A P. multocida mesA mutant was generated which resulted in the loss of acetyl esterase activity under anaerobic conditions. However, this mutation did not cause any attenuation of virulence for mice nor a detectable change to the anaerobic hemolytic phenotype of P. multocida. In E. coli MesA appeared to cause hemolysis indirectly by the induction of the latent E. coli K 12 cytolysin, sheA. PMID- 11064204 TI - On the mechanism of resistance to channel-forming colicins (PacB) and tellurite, encoded by plasmid Mip233 (IncHI3). AB - Plasmids of the H incompatibility complex confer protection against all known channel-forming colicins (PacB character) and resistance to potassium tellurite (Te(r)) to Escherichia coli strains. A DNA clone (2.2 kbp) from plasmid Mip233 (IncHI3) expressing PacB-Te(r) phenotypes was studied. DNA sequence analysis revealed a high degree of homology with the enzyme O-acetylserine sulfhydrylase. Size of the PacB-Te(r) transcript was estimated as 1200 bases. A single polypeptide was found on SDS-polyacrylamide gel with a molecular mass estimated of 34 kDa. The effect of channel-forming colicins and tellurite was analyzed at physiological and transcriptional levels. Results suggest that the pacB gene product could be a reductase-like enzyme. It is also suggested that presence of the PacB character among H plasmid confers selective advantage on cells sharing an ecological niche. PMID- 11064205 TI - Transcriptional analysis of inducible acetamidase gene of Mycobacterium smegmatis. AB - Isolation and analysis of strong and regulatable promoters of mycobacteria should be useful tools to aid the expression of cloned genes in mycobacteria. In the present study, we have mapped the transcriptional start site of the inducible acetamidase gene of Mycobacterium smegmatis and studied the mechanism of its regulation. Northern blot, reverse transcription-PCR and primer extension analysis studies were used to determine the position of the promoter. PMID- 11064206 TI - Characterization of the Calu-3 cell line as a tool to screen pulmonary drug delivery. AB - The objective of this research was to examine the human sub-bronchial gland cell line, Calu-3, and assess its potential as a metabolic and transport model to study drug delivery to the respiratory epithelium. The present studies were conducted using Calu-3 cells grown in Transwells(R) or in multiwell cluster plates. TEER values for Calu-3 monolayers were determined using the World Precision Instrument Voltohmmeter and STX-2 electrode. The results confirmed that Calu-3 cells form tight monolayers and give appreciable TEER values in culture when grown under air-interface conditions. Permeability data for small lipophilic molecules across Calu-3 monolayers suggested that the cell line is a suitable model to examine the transport of low molecular weight substances and xenobiotics. Calu-3 cells were also found to efflux FITC-transferrin (MW 80000) in a polarized manner. The metabolic capacity of Calu-3 cells was also examined. The P4501A1 and P4502B isozymes were determined to be functional, but not inducible, with fluorescent resorufin assays. The data indicated that the Calu-3 cell line may be useful for studying the contributions of bronchial epithelial cells to mechanisms of drug delivery at the respiratory epithelium. PMID- 11064207 TI - Effect of aging on the release of salbutamol sulfate from lipid matrices. AB - The aim of the present work was to evaluate the influence of aging that might condition the release of salbutamol sulfate from oral formulations (lipid matrices) using Gelucire as lipid excipients. Gelucires are essentially characterized by their melting point and their hydrophilic-lipophilic balance. The release profiles of salbutamol sulfate from the capsules elaborated were dependent on the type of Gelucires, fast release, in the case of Gelucire 35/10, a slower release for Gelucire 48/09 and a slow release for Gelucire 46/07. Differential scanning calorimetry was used to study the physical state of drugs in the matrices. Gelucires may exhibit aging effects, whereby a range of physical properties may change upon storage. In the case of Gelucire 35/10, which presents a fast release of salbutamol sulfate, storage produces a decrease in the values of dissolution constant for all capsule sizes. Gelucire 48/09 showed a slower release rate than Gelucire 35/10, and after 1 year of storage, a decrease in the salbutamol dissolution rate for capsule number 3 and 4 was observed. Gelucire 46/07 presented the slowest dissolution rate, but there were not statistically significant differences. These results show that the faster the dissolution rate, and the larger the capsule size, the higher is the influence of storage. PMID- 11064208 TI - Route of decomposition of thiomersal (thimerosal). AB - The route of formation and identification of the principal degradation products of thimerosal (thiomersal) has been undertaken. The initial oxidation to dithiosalicylic acid is followed by cleavage of the disulphide bond of the dithiosalicylic acid by the ethylmercuric ion to reform 1.5 mol of thimerosal with concurrent oxidation to form 0.5 mol of 2-sulfinobenzoic acid for each mole of dithiosalicylic acid. In the presence of copper ions 2-sulfobenzoic acid was also formed. A mechanism has been proposed which accounts for the stoichiometry of the cleavage reaction observed and the significance of the reaction is discussed. PMID- 11064209 TI - Permeability of antisense oligonucleotide through porcine buccal mucosa. AB - Antisense oligonucleotides (AONs) that can modulate malfunctioning genes have a great potential to become future therapeutic agents. In this study, we investigated the feasibility of buccal delivery of AONs using ISIS 3082 as a model compound. An isocratic HPLC method was developed to quantify ISIS 3082. The permeability coefficient of this AON at 37 degrees C, determined by using side-by side diffusion cells, was 1.05x10(-9) (cm/s). The flux of ISIS 3082 across buccal mucosa was dependent upon its concentration in the donor chamber. The permeation of ISIS 3082 was increased when 100 mM of sodium glycocholate was used as a permeation enhancer. The potential of delivering AONs via buccal route with the aid of permeation enhancers is explored in this study. PMID- 11064210 TI - DNA transfection and transfected cell viability using amphipathic asymmetric dendrimers. AB - Amphipathic asymmetric dendrimers have been investigated for use in delivery of genes into cells, with the objective of optimising transfection efficiency and maintaining cell viability. We have synthesised amphipathic asymmetric dendrimers by solid phase methods. The ability of two of these to transfect BHK cells in culture with beta-galactosidase gene was determined by X-gal staining. Cell viability was measured by the MTT assay for BHK cells, and by spectroscopy for lysis of erythrocytes. Interactions between dendrimer and DNA were investigated by agarose gel electrophoresis. BHK cells were optimally transfected at 5:1 +/- charge ratio yielding 20% cells receiving at least one copy of the plasmid. Cell viability decreased when the dendrimer to DNA ratio exceeded 5:1. Raising the pH significantly affected the electrophoretic mobility of complexes of dendrimer and DNA. We conclude that amphipathic asymmetric dendrimers enable efficient plasmid DNA uptake into BHK cells. Cell viability is maintained at high concentrations of dendrimer when complexed with DNA at a 5:1 +/- charge ratio. Efficiency of transfection and cell viability suggest the system may be suitable for gene delivery in vivo. PMID- 11064211 TI - Human lung deposition data: the bridge between in vitro and clinical evaluations for inhaled drug products? AB - Regulatory dossiers for new inhaled drug products generally contain in vitro data, which assess delivered dose and particle size distribution, together with clinical efficacy and safety data. Human lung deposition data may be generated using radionuclide imaging techniques or appropriate pharmacokinetic methods, and can act as a 'bridge' via which a seamless transition can be made between in vitro testing in the laboratory and efficacy/safety testing in the clinic. By enabling informed decisions to be made about the evaluation of new devices or formulations in man, lung deposition data permit a long and expensive clinical trials programme to be commenced with much greater certainty of a successful outcome. Human lung deposition data should be considered for supplementing the information required for regulatory dossiers. PMID- 11064212 TI - A unique dosage form to evaluate the mechanical destructive force in the gastrointestinal tract. AB - The purpose of this study was to prepare tablets that could evaluate the destructive force in the gastrointestinal (GI) tract. Many factors are known to affect in vivo drug release from oral dosage forms. There is still relatively little information on the mechanical destructive force in the GI tract. Press coated tablets with an extremely brittle outer layer were developed using a unique, highly hydrophobic Teflon powder that could be shaped with weak compression force. A marker drug contained in the tablets was released only when the tablets received a force larger than its predetermined crushing strength. We referred to this type of tablet as a 'destructive force dependent release system' (DDRS). A total of nine healthy, male subjects were orally administered the tablets under fed and/or fasting conditions. Tablets with a predetermined crushing strength of 1.50 N were crushed by all of the four subjects who took them under fed conditions and two of the five subjects under fasting conditions. Tablets with a crushing strength of 1.89 N were crushed by two of the six subjects who took them under fed conditions and none of the five subjects under fasting conditions. The range of mechanical destructive force in the human stomach was obtained. PMID- 11064213 TI - Colloidal carriers for benzathine penicillin G: nanoemulsions and nanocapsules. AB - The main purpose of this work is to formulate benzathine penicillin G nanoemulsion and nanocapsules, to evaluate their physicochemical and stabilising characteristics, and to determine their antimicrobial activity and penicillin in vitro release kinetics. Nanoemulsions were produced by the spontaneous emulsification approach and nanocapsules of poly (D,L-lactic acid-co-glycolic acid) polymer (PLGA) were prepared by the method of interfacial deposition of a pre-formed polymer. A 207+/-8 nm mean diameter nanoemulsion formulation maintained stability for more than 5 months at 4 degrees C. Stable nanocapsules with 224+/-58 nm mean diameter were obtained, which remained stabilised over 120 days at 4 degrees C. The penicillin encapsulation ratio in the nanocapsules was 85%. The in vitro release profiles indicated that penicillin released from the nanoemulsion was similar to the one observed from nanocapsules. However it can be clearly deduced from the in vitro kinetic analysis that the antibiotic cannot be protected in colloidal delivery systems. Nevertheless, stable formulations obtained in this investigation supply a potential dosage form to encapsulate more easily soluble drugs. PMID- 11064214 TI - Highly compressible paracetamol: I: crystallization and characterization. AB - It was found that polyvinylpyrrolidone (PVP) is an effective additive during crystallization of paracetamol and significantly influenced the crystallization and crystal habit of paracetamol. These effects were attributed to adsorption of PVP onto the surfaces of growing crystals. It was found that the higher molecular weights of PVP (PVP 10000 and PVP 50000) were more effective additives than lower molecular weight PVP (PVP 2000). Paracetamol particles obtained in the presence of 0.5% w/v of PVP 10000 or PVP 50000 had near spherical structure and consisted of numerous rod-shaped microcrystals which had agglomerated together. Particles obtained in the presence of PVP 2000 consisted of fewer microcrystals. Differential scanning calorimetry (DSC) and X-ray powder diffraction (XPD) experiments showed that paracetamol particles, crystallized in the presence of PVP, did not undergo structural modifications. By increasing the molecular weight and/or the concentration of PVP in the crystallization medium the amount of PVP incorporated into the paracetamol particles increased. The maximum amount of PVP in the particles was 4.32% w/w. PMID- 11064215 TI - Highly compressible paracetamol - II. Compression properties. AB - Paracetamol particles crystallized in the presence of polyvinylpyrrolidone (PVP) exhibited an obvious improvement in their compression properties compared to untreated paracetamol. Paracetamol particles crystallized in the presence of 0.5% w/v PVP 10000 or PVP 50000 produced tablets with improved crushing strength with no tendency to cap even at high compression speeds. The very low values of strain rate sensitivity (SRS) and the lack of reduction in crushing strength with increasing compression speed for these particles, were indicative of a high degree of fragmentation during compression. The results of elastic recoveries and elastic energies of tablets were indicative of much less elastic behaviour of these particles than untreated paracetamol. The low elastic energy/plastic energy (EE/PE) ratio for paracetamol crystallized in the presence of PVP indicated that, contrary to untreated paracetamol, a minor portion of compression energy was utilized as elastic energy. PMID- 11064216 TI - The use of different sugars as fine and coarse carriers for aerosolised salbutamol sulphate. AB - The aim of this study was to investigate the dispersion and deaggregation of a model drug, salbutamol sulphate (SS), using lactose, mannitol or sorbitol as coarse and fine carriers. Binary and tertiary formulations containing micronised salbutamol sulphate (SS) and sieved (63-90 microm) coarse sugar crystals or salbutamol sulphate (SS) with a mixture of coarse and fine sugar particles were prepared. Factorial design was employed to investigate the effects of three variables, i.e. the chemical entity of the coarse sugar carrier, the chemical entity of the fine sugar and the concentration of fine sugar, on the dispersion and deaggregation of salbutamol sulphate after aerosolisation at 60 l/min via a Rotahaler(R) into a twin stage liquid impinger (TSI). The binary formulations containing the different sugar entities produced differences in the fine (<6.4 microm) particle fraction (FPF) of SS in a decreasing order of mannitol >sorbitol >lactose, but failed to produce efficient dispersion of SS since the FPF was <10%. Adding fine sugar particles and increasing their concentration to the binary mixtures generally resulted in an increase in the FPF of salbutamol sulphate. The chemical nature of the fine carriers was found to play a less important role in determining respirable fraction of the drug than the coarse carriers. In conclusion, other sugars such as mannitol or sorbitol, besides lactose, may be employed as coarse and/or fine carriers for incorporation into dry powder aerosol formulations to increase FPF. PMID- 11064217 TI - Induction of ovarian cystic follicles in sheep. AB - Cystic follicles are a significant cause of infertility in women, dairy cattle and sheep. Sheep were used as a model to identify factors that may elicit formation of cystic follicles. Insulin resistance and elevated LH activity were tested in overweight ewes because of associations among these factors and the formation of cystic follicles. Sheep were synchronized using a progesterone releasing pessary and insulin resistance was induced during the synchronization period through administration of bovine somatotropin. Following removal of pessaries follicular growth was stimulated by treatment with eCG or eCG and hCG (PG-600). Follicular growth was monitored via daily transrectal ultrasonography and blood samples were collected for hormonal analyses. Six of 18 ewes had a subnormal or absent preovulatory gonadotropin surge and developed cystic follicles. Neither insulin resistance nor elevated LH activity were associated with formation of cystic follicles. Ewes that developed cystic follicles were heavier (93 +/- 4 kg) than ewes that ovulated (81 +/- 3 kg; P = 0.02). Furthermore, following pessary removal and initiation of daily ultrasonography, ewes that developed cystic follicles lost body weight (-3 +/- 1%), while ovulatory ewes continued to gain body weight (1 +/- 1%; P = 0.005). It is speculated that in heavy ewes metabolic factors associated with acute body weight loss inhibit the positive feedback of estradiol and thereby suppress the preovulatory gonadotropin surge leading to formation of cystic follicles. PMID- 11064218 TI - The involvement of prolactin in the regulation of adrenal cortex function in pigs. AB - We investigated the in vivo and in vitro effect of prolactin (PRL) on porcine adrenal cortex function. The in vivo study was performed on 10 multiparous sows. Blood was sampled every 4 h beginning on the 17th day of the estrous cycle and continuing for 6 subsequent days. Plasma was stored at -20 degrees C until steroid hormones analysis was completed. PRL or saline were administered iv for 48 h in 2 h intervals. Injections of PRL began 4-20 h after the preovulatory LH surge. At the end of the sampling period sows were slaughtered and adrenals were immediately dissected. Adrenals were frozen at -70 degrees C for determination of adrenal cortex steroid hormones content. At the end of PRL treatment period mean plasma level of cortisol in control sows was significantly lower than that of PRL treated sows. Moreover, the area under the mean plasma cortisol concentration curve was significantly higher in PRL-treated sows in comparison to controls. The mean cortisol adrenal content was significantly higher in adrenal cortex of PRL treated sows than that of controls. PRL did not affect adrenal cortex concentration of androstenedione (A(4)), testosterone (T), dehydroepiandrosterone (DHEA) and estradiol (E(2)). Dehydroepiandrosterone sulfate (DHEAS) was not found in porcine adrenal cortex. In the in vitro experiment adrenal glands were removed immediately after slaughter of 6 crossbred gilts. Dispersed adrenocortical cells were incubated for 8 h with or without porcine PRL. Prolactin stimulated cortisol secretion in a dose-dependent manner. These results suggest that PRL is one of the key factors involved in the regulation of adrenal cortex function in pigs. PMID- 11064219 TI - Cholecystokinin mediates depression of feed intake in dairy cattle fed high fat diets. AB - Two experiments were conducted to investigate the effects of 1) different concentrations of dietary fat and 2) i.v. administration of a cholecystokinin receptor antagonist (MK-329) on feed intake and plasma concentrations of hormones and metabolites in dairy cattle. In Experiment 1, 4 lactating Holstein cows were used in a 4 x 4 Latin square design. Treatments were diets with 1) no fat added, 2) 30 g fat/kg feed (calcium salts of long-chain fatty acids as fat supplement), 3) 60 g fat/kg, and 4) 90 g fat/kg added. Cows were fed once daily a diet of concentrate, corn silage, alfalfa haylage, and alfalfa pellets. Dry matter intake decreased linearly with increasing concentrations of dietary fat (P < 0.0001). Overall plasma concentrations of nonesterified fatty acids (P < 0.0001), triacylglycerol (P < 0.0006), and cholecystokinin (P < 0.02), increased linearly with each level of dietary fat, but there was a linear decrease in plasma insulin (P < 0.0008). In Experiment 2, 4 nonpregnant and nonlactating Holstein heifers were used in a cross-over design in a 2 x 2 factorial arrangement of treatments. Treatments were diet (fatty acids, 27 g/kg vs 103 g/kg diet dry matter) and i.v. injections (MK-329 vs vehicle). Heifers were fed once daily a total mixed ration of corn silage, cracked corn and soybean meal with or without fat supplement. Diets were switched by period and either MK-329 (70 microg/kg body weight) or its vehicle was injected i.v. at 2 hr postfeeding. Daily dry matter intake was decreased by feeding the high fat diet (P < 0.02) but was not affected by injections. Injection of MK-329, however, increased dry matter intake by 92% in heifers fed the high fat diet during the first 2 hr postinjection compared to vehicle injection. Plasma pancreatic polypeptide concentration was increased by the high fat diet at 2 hr postfeeding (P < 0.02) but was lowered by MK-329 at 1 hr postinjection (P < 0.001). Plasma insulin was lowered by the high fat diet (P < 0.01) but was not affected by injections. The elevated plasma cholecystokinin concentration may have mediated depressed feed intake of dairy cattle fed the high fat diets. PMID- 11064220 TI - Combined pituitary hormone deficiency in german shepherd dogs with dwarfism. AB - In German shepherd dogs pituitary dwarfism is known as an autosomal recessive inherited abnormality. To investigate whether the function of cells other than the somatotropes may also be impaired in this disease, the secretory capacity of the pituitary anterior lobe (AL) cells was studied by a combined pituitary AL stimulation test with four releasing hormones (4RH test) in four male and four female German shepherd dwarfs. In addition, the morphology of the pituitary was investigated by computed tomography. The physical features of the eight German shepherd dwarfs were primarily characterized by growth retardation and stagnant development of the hair coat. The results of the 4RH test confirmed the presence of hyposomatotropism. The basal plasma TSH and prolactin concentrations were also low and did not change upon stimulation. Basal plasma concentrations of LH were relatively low and responded only slightly to suprapituitary stimulation. With respect to the plasma FSH levels there was a clear gender difference. In the males plasma FSH concentrations remained below the detection limit throughout the 4RH test, whereas in the females the basal plasma FSH levels were slightly lower and there was only a small increase following suprapituitary stimulation, compared with the values in age-matched controls. In contrast, basal and stimulated plasma ACTH concentrations did not differ between the dwarfs and the controls. Computed tomography of the pituitary fossa revealed a normal sized pituitary with cysts in five dogs, an enlarged pituitary with cysts in two dogs, and a small pituitary gland without cysts in the remaining dog. The results of this study demonstrate that German shepherd dwarfs have a combined deficiency of GH, TSH, and prolactin together with impaired release of gonadotropins, whereas ACTH secretion is preserved. The combined pituitary hormone deficiency is associated with cyst formation and pituitary hypoplasia. PMID- 11064221 TI - mRNA of insulin-like growth factor (IGF) quantification and presence of IGF binding proteins, and receptors for growth hormone, IGF-I and insulin, determined by reverse transcribed polymerase chain reaction, in the liver of growing and mature male cattle. AB - Plasma insulin-like growth factor-I (IGF-I) concentrations were related to hepatic levels of IGF-I mRNA measured by competitive reverse transcription polymerase chain reaction (PCR) (RT-PCR) in neonatal (8 d old) calves, veal calves, fattened castrated bulls and mature intact bulls. Furthermore, the presence of mRNAs of IGF-II and of receptors for IGF-I (IGF-IR), growth hormone (GHR) and insulin (IR), as well as mRNAs of IGF binding proteins (IGFBP-1, -2 and -3) were assessed by RT-PCR. Hepatic IGF-I mRNA levels and plasma IGF-I concentrations in veal calves, fattened castrated bulls and in intact bulls were 4 to 8 times higher than in 8-d old calves and were 2 to 3 times higher in calves fed colostrum than in calves fed only milk replacer. Hepatic IGF-I mRNA concentrations were closely correlated (r = 0.92) with plasma IGF-I concentrations, suggesting that hepatic IGF-I production largely determines plasma IGF-I levels. The presence of IGF II, IGF-IR, GHR, IR and IGFBP-1, -2 and 3 mRNA was confirmed in the liver of 8-d old calves and older cattle as well, and among newborn calves their presence was independent of differences in nutrition. In conclusion, the major hepatic components of the GH-IGF axis were present in neonatal calves, but the IGF-I expression and therefore also plasma IGF-I levels were relatively low. PMID- 11064222 TI - Violence against women: charting the impact on health policy, health care delivery, and the law. PMID- 11064223 TI - Predictors of post-rape medical care in a national sample of women. AB - BACKGROUND: Rape has a negative impact on physical and mental health, health related behaviors, and health service utilization. Timely medical care is important for preventive services. METHODS: Cross-sectional data were obtained from a larger 2-year longitudinal study, the National Women's Study (NWS). A total of 3006 adult women participated in the final data collection wave of the NWS. During a structured telephone interview, women who reported a most-recent or only rape incident during adulthood were asked about rape characteristics, reporting to authorities, medical care, and rape-related concerns. The main outcome measures were receipt and timing of medical care received after an adult rape, and factors influencing whether or not medical care was received. RESULTS: Of the sample, 214 (7.1%) had experienced a most-recent or only rape as an adult (aged >/=18), and 56 (26.2%) received rape-related medical care following that incident. The final model multivariable logistic regression indicated that reporting the crime to police or other authorities (odds ratio [OR], 9.45; 95% confidence interval [CI]=3. 34-26.70) and fear of sexually transmitted diseases (OR, 8.61; 95% CI=3.12-23.72) were significant predictors of receipt of post-rape medical care. CONCLUSIONS: One in five victims reported an adult rape to police or other authorities; these women were nine times more likely to receive medical care than those who did not. Public health efforts are needed to increase the proportion of rape victims who receive immediate post-rape medical care. PMID- 11064224 TI - Severe dating violence and quality of life among south carolina high school students. AB - BACKGROUND: Little research has addressed the impact of dating violence and forced-sex victimization and perpetration on adolescent well-being. In this cross sectional study, we provide (1) estimates of severe dating violence (SDV) by victimization and perpetration status, (2) estimates of lifetime forced-sex victimization and perpetration, (3) demographic and health behaviors correlated with SDV, and (4) associations between SDV and forced sex and well-being as assessed by (1) health-related quality of life (H-R QOL) and (2) life satisfaction measures. METHODS: We used a stratified cluster sample of 5414 public high school students, grades 9 through 12, who responded to the 1997 self administered South Carolina Youth Risk Behavior Survey. RESULTS: Nearly 12% of adolescents self-reported SDV as a victim (7.6%) or a perpetrator (7.7%), and SDV rates (victimization/perpetration combined) are higher in girls (14.4%) than boys (9.1%). Race, aggressive behaviors, substance use, and sexual risk-taking are correlates of SDV. Among young women, SDV victimization, not perpetration, was associated with recent poor H-R QOL and suicide ideation or attempts, but not lower life-satisfaction scores. Among young men, SDV perpetration, not victimization, was strongly associated with poor H-R QOL and suicide attempts, and lower scores for all domains of life satisfaction. CONCLUSIONS: This research provides evidence that SDV and forced sex are associated with poor H-R QOL, low life-satisfaction scores, and adverse health behaviors in adolescent female victims and male perpetrators. Screening for dating violence is needed to identify and intervene early to reduce the impact of dating violence. PMID- 11064225 TI - Prevention of pregnancy resulting from rape: a neglected preventive health measure. AB - Abstract: Pregnancy following rape is a continuing and significant public health issue. We estimate that the 333,000 sexual assaults and rapes reported in 1998, along with many more unreported, were responsible for 25,000 pregnancies. Potentially, as many as 22,000 such pregnancies could be prevented if all women who were raped received prompt medical services, and if not already protected against pregnancy, were provided with emergency contraceptive treatment. PMID- 11064226 TI - Screening for intimate partner violence by health care providers. Barriers and interventions. AB - INTRODUCTION: Routine screening for intimate partner violence (IPV) is endorsed by numerous health professional organizations. Screening rates in health care settings, however, remain low. In this article, we present a review of studies focusing on provider-specific barriers to screening for IPV and interventions designed to increase IPV screening in clinical settings. METHODS: A review of published studies containing original research with a primary focus on screening for IPV by health professionals was completed. RESULTS: Twelve studies identifying barriers to IPV screening as perceived by health care providers yielded similar lists; top provider-related barriers included lack of provider education regarding IPV, lack of time, and lack of effective interventions. Patient-related factors (e.g., patient nondisclosure, fear of offending the patient) were also frequently mentioned. Twelve additional studies evaluating interventions designed to increase IPV screening by providers revealed that interventions limited to education of providers had no significant effect on screening or identification rates. However, most interventions that incorporated strategies in addition to education (e.g., providing specific screening questions) were associated with significant increases in identification rates. CONCLUSION: Barriers to screening for IPV are documented to be similar among health care providers across diverse specialties and settings. Interventions designed to overcome these barriers and increase IPV-screening rates in health care settings are likely to be more effective if they include strategies in addition to provider education. PMID- 11064227 TI - Opportunities for intervention: discussing physical abuse during prenatal care visits. AB - BACKGROUND: The American Academy of Pediatrics and the American College of Obstetricians and Gynecologists recommend that screening for physical abuse during prenatal care visits becomes routine. Although prenatal care visits offer a unique intervention opportunity, screening is not yet standard practice. DATA AND METHODS: We used data from the 1996 and 1997 Pregnancy Risk Assessment Monitoring System (PRAMS) to assess the prevalence of and the factors associated with health care providers' discussion of physical abuse with pregnant women in 14 states. PRAMS is a state-specific, population-based surveillance system that collects information from women on maternal behaviors before and during pregnancy, and at 2 to 6 months postpartum. RESULTS: Between 22% and 39% of the women surveyed reported that health care providers talked with them about physical abuse during prenatal care visits. Health care providers were more likely to discuss physical abuse with women who were black, Hispanic, young (aged <20 and 20 to 29), had a high school education or less, or paid for prenatal care with Medicaid. CONCLUSIONS: Our results indicate that most pregnant women do not report that their prenatal care providers discussed physical abuse with them. Logistic regression analyses identified consistent associations across the 14 states between discussion of abuse and demographic and pregnancy-related factors. A better understanding of the factors associated with whether a health care provider discusses physical abuse with a pregnant woman could increase intervention opportunities. PMID- 11064228 TI - Development of a health care provider survey for domestic violence: psychometric properties. AB - BACKGROUND: Despite rapid proliferation of descriptive studies of health care providers (HCPs) and protocols for identification and management of domestic violence (DV), few reliable instruments exist for assessing HCPs' attitudes, beliefs, and behaviors regarding this practice. This study describes the development and psychometric properties of a measure of attitudes, beliefs, and self-reported behaviors related to the identification and management of DV. METHODS: We used a multiphase study design to develop items across eight content domains. We administered an initial pool of 104 items to a pilot sample of 129 primary care providers (physicians, physician assistants, nurse practitioners, and medical assistants) in a large, urban health maintenance organization. Descriptive statistics, principal components, and reliability analyses were performed on each of the eight content domains. The analyses guided the deletion of items and development of additional items, yielding a 56-item pool. The items were then administered and re-analyzed with an independent sample of 246 HCPs. RESULTS: Six separate and reliable domains were identified: Perceived Self Efficacy, System Support, Blame Victim, Professional Role Resistance/Fear of Offending Patient, Victim/Provider Safety, and Frequency of DV Inquiry. We found item domain Cronbach alpha to be acceptable, ranging from 0.73 to 0.91. The final overall measure had 39 items and an alpha of 0.88. Data are reviewed that support the measure's sensitivity to change in response to a training intervention. CONCLUSION: The measure provides a reliable method for assessing provider characteristics and training needs. It may also serve to evaluate training and policy interventions in DV. PMID- 11064229 TI - Identification and management of domestic violence: a randomized trial. AB - BACKGROUND: Diagnosis of domestic violence (DV) in primary care is low compared to its prevalence. Care for patients is deficient. Over a 1-year period, we tested the effectiveness of an intensive intervention to improve asking about DV, case finding, and management in primary care. The intervention included skill training for providers, environmental orchestration (posters in clinical areas, DV questions on health questionnaires), and measurement and feedback. METHODS: We conducted a group-randomized controlled trial in five primary care clinics of a large health maintenance organization (HMO). Outcomes were assessed at baseline and follow-up by survey, medical record review, and qualitative means. RESULTS: Improved provider self-efficacy, decreased fear of offense and safety concerns, and increased perceived asking about DV were documented at 9 months, and also at 21 months (except for perceived asking) after intervention initiation. Documented asking about DV was increased by 14.3% with a 3.9-fold relative increase at 9 months in intervention clinics compared to controls. Case finding increased 1.3 fold (95%, confidence interval 0.67-2.7). CONCLUSIONS: The intervention improved documented asking about DV in practice up to 9 months later. This was mainly because of the routine use of health questionnaires containing DV questions at physical examination visits and the placement of DV posters in clinical areas. A small increase in case finding also resulted. System changes appear to be a cost effective method to increase DV asking and identification. PMID- 11064230 TI - Partner violence and medical encounters: African-American women's perspectives. AB - OBJECTIVE: To examine the relationship between intimate partner violence (IPV) victimization and patient satisfaction with medical encounters among an African American population. DESIGN: Cross-sectional, self-administered, anonymous survey. SETTING: Community-based, primary care center. PATIENTS: Consecutive African-American women recruited from an urban health center. A total of 102 women provided sufficient information to reveal whether they were currently experiencing IPV and to allow us to assess their experiences in their most recent primary care encounter. MEASUREMENTS: Patients' perceptions of their most recent encounter using questions adopted from the Medical Interview Satisfaction Scale and Consultation Satisfaction Questionnaire. We used the Conflicts Tactics Scale, supplemented with questions measuring sexual violence and emotional abuse, to assess IPV "in the past year." RESULTS: Women who reported current IPV rated several aspects of the encounter more negatively than did women who did not report current abuse. The IPV victims were less likely to report that they felt respected and accepted during the encounter, and they provided lower ratings of the quality of communication with their providers. CONCLUSIONS: It is unclear why victims of partner violence experience medical encounters as less satisfactory. Researchers need to expand studies of medical encounters as experienced by abused women to determine whether IPV status adversely affects general medical care. PMID- 11064231 TI - Adverse consequences of intimate partner abuse among women in non-urban domestic violence shelters. AB - OBJECTIVE: This study examined the health consequences of having experienced both sexual and physical abuse relative to women experiencing physical abuse but not sexual abuse. METHODS: A cross-sectional study was conducted among 203 women seeking refuge in battered women's shelters. Controlling for sociodemographics, logistic regression analyses were conducted to assess the consequences of experiencing both sexual and physical abuse. RESULTS: Compared to women experiencing physical abuse, women experiencing both sexual and physical abuse were more likely to have a history of multiple sexually transmitted diseases (STDs) in their abusive relationships, have had an STD in the past 2 months, be worried about being infected with HIV, use marijuana and alcohol to cope, attempt suicide, feel as though they had no control in their relationships, experience more episodes of physical abuse in the past 2 months, rate their abuse as more severe, and be physically threatened by their partner when they asked that condoms be used. CONCLUSIONS: Given the prevalence of adverse health outcomes, domestic violence shelters could counsel women to avoid using alcohol/drugs as a coping strategy, educate women about alternative healthy coping strategies, counsel women about methods of STD prevention that they can control, and provide STD screening and treatment. PMID- 11064232 TI - Women's opinions about domestic violence screening and mandatory reporting. AB - BACKGROUND: The purpose of this paper is to describe women's opinions and policy preferences concerning domestic violence screening and mandatory reporting. METHODS: This case-control study included 202 abused women and 240 randomly selected non-abused women recruited from a large metropolitan health maintenance organization who were interviewed by telephone. Of these women, 46.6% had a college degree, 53.4% were white, and 60% had a household income of $50,000 or more. RESULTS: Forty-eight percent of the sample agreed that health care providers should routinely screen all women, with abused women 1.5 times more likely than non-abused women to support this policy. For mandatory reporting, 48% preferred that it be the woman's decision to report abuse to the police. Women thought it would be easier for abused women to get help with routine screening (86%) and mandatory reporting (73%), although concerns were raised about increased risk of abuse with both screening (43%) and reporting (52%) policies. Two thirds of the sample thought women would be less likely to tell their health care providers about abuse under a mandatory reporting policy. Interventions offered in managed care settings that would be well received, according to the women in this study, include counseling services, shelters, and confidential hotlines. CONCLUSIONS: Women expressed fears and concerns about negative consequences of routine screening and, even more so, for mandatory reporting. Domestic violence policies and protocols need to address the safety, autonomy, and confidentiality issues that concern women. PMID- 11064233 TI - Who gets protection orders for intimate partner violence? AB - BACKGROUND: It is unknown how victims of intimate partner violence (IPV) who seek civil protection orders differ from IPV victims who do not. METHODS: To compare characteristics of women with and without protection orders, 448 women with police or court contact for an IPV incident in Seattle, Washington, were interviewed. Data collected included demographic characteristics of the subject and her abuser, abuse history, and the subject's mental and physical health. RESULTS: IPV victims who obtained protection orders were more likely than victims without protection orders to be employed full-time, be pregnant, be married, aged over 24, and less likely to be involved with perpetrator at index incident. The perpetrators for both groups were similar, and the majority had a current or previous alcohol/drug problem and a previous criminal history. Both groups of victims had been psychologically and physically abused during the previous year and nearly all had symptoms of depression. However, at the index incident, women who sought protection orders were less likely to be physically assaulted or injured, but more likely to have family members or friends physically assaulted. CONCLUSIONS: Financial independence and abuse of family or friends are important factors associated with the decision to seek a protection order in IPV. PMID- 11064234 TI - Public policy for violence against women: 30 years of successes and remaining challenges. PMID- 11064235 TI - Health status and health care use of Massachusetts women reporting partner abuse. AB - BACKGROUND: Studies indicate that women abused by their intimate partners are at increased risk for a number of health problems and have increased rates of health care utilization. However, these findings are based mainly on studies using clinic or health plan populations. In this study, we examined the association between intimate partner abuse (IPA) and health concerns and health care utilization in a population-based sample of adult women. METHODS: We analyzed data on 2043 women aged 18 to 59 who participated in the 1998 Massachusetts Behavioral Risk Factor Surveillance System (BRFSS), a population-based health survey that included questions on IPA. IPA was defined as experiencing physical violence by, fear of, or control by an intimate partner. Consequences of IPA and self-rated health status and health care utilization of women experiencing IPA were examined. RESULTS: A total of 6.3% of Massachusetts women aged 18 to 59 reported IPA during the past year. Women experiencing IPA were more likely than other women to report depression, anxiety, sleep problems, suicidal ideation, disabilities, smoking, unwanted pregnancy, HIV testing, and condom use. Women experiencing IPA were less likely to have health insurance, but received routine health care at similar rates as other women. CONCLUSIONS: These results indicate that women in the general population experiencing IPA are at increased risk for several serious emotional and physical health concerns. Most of these women are in routine contact with health care providers. These findings also suggest that the BRFSS may provide a valuable mechanism for tracking state-based IPA prevalence rates over time. PMID- 11064236 TI - Improving surveillance of intimate partner violence by use of multiple data sources. AB - BACKGROUND: Intimate partner violence (IPV) is a significant public health problem in the United States. Estimates of incidence and prevalence vary widely, depending on the data source used. Combining information from different sources can enhance our understanding of IPV. METHODS: In this paper, we used 1998 data from the Rhode Island (RI) Department of Health Violence Against Women Public Health Surveillance System to describe the prevalence of IPV reported to police, the demographic characteristics and help-seeking efforts of women reporting IPV, and characteristics of IPV incidents. We used data from the 1998 RI Department of Health Behavioral Risk Factor Surveillance System survey to examine associations between health care use and health outcomes of victims and nonvictims of IPV, and to explore the correlates of IPV. We also discuss the use of both narrow and broad definitions of IPV. RESULTS: Our findings show that the definition of IPV and the source used to identify IPV victims can produce a markedly different picture of IPV victims, and that combining information from different data sources can enhance our understanding of IPV. An important finding for health care providers is that IPV victims do not appear to be significantly different from nonvictims in their access to and utilization of routine health care, and that more than 60% of victims at highest risk for injury reported seeing a health care provider because of IPV. CONCLUSIONS: Our findings underscore the importance of health care providers addressing IPV and its consequences among their patients. PMID- 11064237 TI - Help-seeking for intimate partner violence and forced sex in South Carolina. AB - PURPOSE: In this population-based, random-digit-dial, cross-sectional survey, we assessed the lifetime victimization of intimate partner violence (IPV) and forced or coerced sex among 556 women and men in South Carolina, and the help-seeking behaviors of victims. RESULTS: Among women, 25.3% experienced IPV (sexual, physical, or emotional violence) compared with 13.2% of men. Although women were significantly more likely to report physical or sexual IPV (17.8%) than were men (4.9%), men (8.3%) were as likely as women (7.4%) to report perceived emotional abuse without physical or sexual IPV. One half of men and women with annual incomes <$15, 000 reported IPV. Among women experiencing physical or sexual IPV, 53% sought community-based or professional services for IPV; women with higher education levels and those experiencing more severe violence were most likely to seek services. CONCLUSIONS: These data show that IPV is common and that most victims do not receive services to address this violence. PMID- 11064238 TI - Population surveillance for physical violence among adult men and women, Montana 1998. AB - BACKGROUND: Few epidemiologic studies of physical violence or intimate partner violence provide population-based surveillance data. OBJECTIVES: To estimate the prevalence and describe the characteristics associated with physical violence among adult men and women in the past year. METHODS: A random sample of Montana households was contacted via the Behavioral Risk Factor Surveillance System telephone survey in 1998 (N=1804). RESULTS: Five percent of men (39/787) and 3% of women (33/1017) reported experiencing physical violence in the past year. Among respondents reporting physical violence in the past year, women were more likely than men to report that the perpetrator was a current/former partner (58% vs 10%, p/=0.05). Men who reported experiencing physical violence in the past year were more likely to be younger and not to be living with a current partner. Women who reported experiencing physical violence in the past year were more likely to be younger, not currently living with a partner, have no health insurance, and have more days with mental health problems in the past month. CONCLUSIONS: Recent physical violence is common for both men and women; however, the perpetrators, locations, and demographic characteristics differ. Further study is needed to better understand the factors associated with physical violence among men and women in the context of designing and implementing appropriate interventions to reduce violence. PMID- 11064239 TI - Violence against women as a public health issue: comments from the CDC. PMID- 11064240 TI - AJPM welcomes new editor for annals of epidemiology PMID- 11064241 TI - History of the Japanese Society for Investigative Dermatology (II) 1991-1999. PMID- 11064242 TI - Comprehensive outlook of in vitro tests for assessing skin irritancy as alternatives to Draize tests. AB - In vitro alternative methods have been verified for the possibility to assess cutaneous irritancy because humans cannot be direct initial experimental subjects and animal experimentation could be forbidden in the near future. Many kinds of cell cytotoxicity assays have been tried, revealing their own advantages and limitations. Cell function-based tests have been used less frequently than cytotoxicity assays. Three-dimensional culture systems are promising because they are closer to the actual in vivo skin, and some of them are commercialized nowadays. The ultimate objective of in vitro irritancy tests, which is the high degree of correlation with human in vivo test results, has been accomplished in many experimental settings. Before applying these in vitro methods we must consider several points, including cell sources, irritant characteristics, exposure time, endpoint of experiment, extrinsic factors affecting irritation, etc. In vitro skin irritancy tests have been developed continuously, and in the future they could assume a heavy responsibility of estimating the irritancy in human skin in vivo. PMID- 11064243 TI - Induction of Ro/SSA antigen expression on keratinocyte cell membrane by heat shock and phorbol 12-myristate 13-acetate as well as estradiol and ultraviolet B. AB - Skin is one of the main target organs in lupus erythematosus and in some circumstances, skin lesions precede systemic manifestations. Previous studies have demonstrated that Ro/SSA antigen antibody might be involved in the pathogenesis of lupus erythematosus. The present study was performed to investigate the factors regulating expression of Ro/SSA antigens on the cell surface of keratinocytes. Cultured normal human keratinocytes were treated with 50-200 mJ/cm(2) of ultraviolet B (UVB) irradiation, 10(-9) to 10(-5) mol/l of 17beta-estradiol, 5-10 microg/l of phorbol 12-myristate 13-acetate (PMA), and 42 and 45 degrees C heat shock, respectively. The Ro/SSA antigen expressions were determined by indirect immunofluorescence. The results showed that keratinocytes receiving UVB irradiation expressed Ro/SSA antigen on cell membranes in a dose dependent fashion. 17beta-estradiol treatment also induced Ro/SSA antigen expression dose-dependently. Keratinocyte expression of Ro/SSA antigens was also induced by heat shock stimulation. The 45 degrees C heat shock showed a stronger effect than 42 degrees C heat shock. Keratinocytes incubated for 24 h after heat shock had more antigen-expressing cells than those incubated for 6 h after heat shock. PMA at 5 and 10 microg/l also strongly induced Ro/SSA antigen expression. These results suggest that Ro/SSA antigen expression can be regulated by many factors and that protein kinase C signal transduction pathway might be involved in this process. PMID- 11064244 TI - Hypoxic conditions decrease the mRNA expression of proalpha1(I) and (III) collagens and increase matrix metalloproteinases-1 of dermal fibroblasts in three dimensional cultures. AB - The effect of hypoxia on the expression of extracellular matrix-related genes by human dermal fibroblasts was investigated using a novel three-dimensional culture supplemented with L-ascorbic acid 2-phosphate. Experiments were performed by placing replicate dishes in either hypoxic (2%) or in normoxic (20%) condition for various periods of time ranging up to 72 h. The mRNA expression levels of proalpha1(I), proalpha1(III) collagens and MMP-1 were analyzed using Northern blotting. Hypoxia transiently increased proalpha1(I) and proalpha1(III) collagen gene expression at 24 h, but a prolonged exposure to hypoxia decreased them. A slight increase in MMP-1 mRNA was observed at 24 h and prolonged exposure for up 72 h resulted in significantly increased expression of MMP-1 gene. Our results suggest that enhanced degradation as well as decreased synthesis of collagens induced by hypoxia may account for the delayed wound healing associated with circulatory disturbances. PMID- 11064245 TI - Epidermal change can alter mechanical properties of hairless mouse skin topically treated with 1alpha, 25-dihydroxyvitamin D(3). AB - Wrinkle formation caused by photoaging clearly involves changes of extracellular matrix components and mechanical properties of the skin. Recently, it was reported that the topical application of 1alpha, 25-dihydroxyvitamin D(3) to hairless mouse skin induced wrinkling. Here we have evaluated the effect of topical application of 1alpha, 25-dihydroxyvitamin D(3), which causes skin wrinkling, on the mechanical properties of the hairless mouse (HR/ICR) skin, using a commercially available non-invasive in vivo instrument. The elasticity element of the skin was unchanged, but the viscosity element significantly increased. Histologically, the epidermis became remarkably thick, but no conspicuous changes were observed in the dermis. Changes in the mechanical properties of the skin after 1alpha, 25-dihydroxyvitamin D(3) treatment take place through epidermal physical variation, especially changes of viscosity elements. It is suggested that the visco-elastic properties of the epidermis are also attributable to the morphology as well as the mechanical properties of the skin. PMID- 11064246 TI - Antimicrobial effects of acidic hot-spring water on Staphylococcus aureus strains isolated from atopic dermatitis patients. AB - The present study examined the antimicrobial effects of acidic hot-spring water on Staphylococcus aureus strains isolated from atopic dermatitis (AD) patients. Plasma coagulation by S. aureus cells was not detected in plasma containing acidic hot-spring water (60%, pH 5.4) or hydrochloric acid (pH 5.0) after incubation for 24 h. S. aureus cells did not grow in Mueller-Hinton broth with acidic hot-spring water (50%, pH 4.4) after 24 h incubation. The colony counts of S. aureus cells in tryptic soy broth containing acidic hot-spring water (60%, pH 3.9) were over ten times lower than those in tryptic soy broth alone after incubation for 24 h (P<0.01). A membranous structure (an immature biofilm) was formed on the coverslips of tissue culture dishes by S. aureus cells in plasma after incubation for 24 h, although the colony counts of S. aureus cells in the immature biofilms in plasma containing acidic hot-spring water (60%, pH 5.4) were about eight times lower than those in plasma alone after incubation for 24 h (P<0.01). The colony counts of S. aureus cells that attached on coverslips in plasma containing acidic hot-spring water (60%, pH 5.4) or hydrochloric acid (pH 5.4) were over 1000 times lower than those in plasma alone after incubation for 24 h. These results suggest that 50% acidic hot-spring water has a bacteriostatic effect, 60% acidic hot-spring water has a moderate bactericidal effect against floating S. aureus cells and those cells in a biofilm, and, 60% acidic hot-spring water has an inhibitory effect on plasma coagulation and attachment of S. aureus cells. Furthermore, our present results suggest that a small amount of some ions in hot-spring water such as manganese and iodide ions are very important for a bactericidal activity of hot-spring water as well as the low pH condition. PMID- 11064247 TI - Analyses of the transglutaminase 1 gene mutation and ultrastructural characteristics in a Japanese patient with lamellar ichthyosis. AB - We described a Japanese female with lamellar ichthyosis whose transglutaminase 1 gene (TGM1 gene) was mutated. DNA sequence analysis revealed that the patient had a homozygous mutation, i.e. a point mutation from G to A at nucleotide 1494 resulting in the substitution of glycine for arginine at codon 143. Her mother was heterozygous for this mutation. In situ transglutaminase assay in the patient's skin showed loss of enzyme activity. Ultrastructural examination revealed incomplete formation of cornified cell envelopes and electron-dense materials adjacent to plasma membranes. These results suggest that defective transglutaminase activity caused by homozygous TGM1 gene mutation (G143R) results in disruption of cornified envelope assembly and the clinical phenotype of lamellar ichthyosis. PMID- 11064248 TI - Treatment of murine angiosarcoma with etoposide, TNP-470 and prednisolone. AB - To develop effective therapies for angiosarcoma, we investigated the anti-tumor effects of etoposide (ETO), TNP-470 and prednisolone (PSL) using an established murine angiosarcoma cell line (ISOS-1). We examined the direct anti-tumor and anti-angiogenic effects of these drugs on ISOS-1 cells and normal murine microvascular endothelial cells (mECs) in vitro. Cell growth of ISOS-1 was inhibited significantly by ETO, moderately by TNP-470, and not at all by PSL (IC(50): 0.25 microg/ml, 10 microg/ml, >8000 microg/ml, respectively). One the other hand, cell growth of mECs was inhibited significantly by TNP-470, slightly by PSL, and negligibly by ETO (IC(50): 0.85 ng/ml, 0.7 microg/ml, 10 microg/ml, respectively). In an in vivo assay, tumor growth of ISOS-1 was significantly inhibited by more than 2.5 mg/kg of ETO dose-dependently, and by more than 30 mg/kg of TNP-470, and 100 mg/kg of PSL individually. Combination treatments of ETO+TNP-470 and TNP-470+PSL showed synergistic enhancement of inhibition (% control inhibition: ETO vs. TNP-470 vs. ETO+TNP-470: 55 versus 55 vs. 16%) (% control inhibition: TNP-470 vs. PSL vs. TNP-470+PSL: 41 vs. 86 vs. 21%). ETO+PSL combination treatment, however, failed to show significant enhancement of anti tumor effects. In conclusion, our results indicated that TNP-470 may be a very effective drug for angiosarcoma treatment, especially in combination with ETO or PSL. We eagerly anticipate the use of TNP-470 in clinical treatment of angiosarcoma. PMID- 11064249 TI - The activity of fatty acid synthase of epidermal keratinocytes is regulated in the lower stratum spinousum and the stratum basale by local inflammation rather than by circulating hormones. AB - The epidermal keratinocytes produce and secrete lipids to maintain the water barrier of the epidermis. To clarify the regulation of epidermal lipid synthesis, we investigated the hormonal effect on the activity of fatty acid synthase (FAS) of the keratinocytes, and the expression of FAS in the human skin. In cultured keratinocytes, the FAS activity, assayed by measuring the oxidation of NADPH, was slightly increased by hydrocortisone or testosterone, but not influenced by thyroid hormone, estrogen, progesterone or insulin. In immunohistochemical study of normal human epidermis, FAS was expressed strongly in the stratum granulosum and moderately in the uppermost layer of the stratum spinousum (SS), suggesting that fatty acid synthesis may increase during normal epidermal differentiation. In inflammatory disorders, such as psoriasis, lichen planus, and atopic dermatitis, FAS was also expressed in the lower SS and the stratum basale (SB), resulting in strong staining in the whole layers of the epidermis. Remarkable increase of FAS expression was only observed in the lower SS and the SB. Therefore, the activity of FAS in the epidermis may be regulated in the lower SS and the SB by local inflammation rather than by circulating hormones. In other components of the skin, FAS was strongly expressed not only in adipose tissue and sebaceous glands, which are known as active sites of lipid synthesis, but also in sweat glands, suggesting that the sweat glands can synthesize abundant fatty acids de novo. PMID- 11064250 TI - The production of superantigenic exotoxins by coagulase-negative staphylococci isolated from human skin lesions. AB - We examined the production of superantigenic exotoxins in 136 coagulase-negative staphylococci isolated from various skin lesions in humans using a reversed passive latex agglutination test (Denka Seiken). As a control we examined the same in 50 Staphylococcus aureus strains isolated from non-infective skin ulcers in humans. Of the 136 strains of coagulase negative-staphylococci, 9 (6.6%) produced one or more identifiable exotoxins. In contrast, 21 (42%) out of the 50 S. aureus strains produced one or more identifiable exotoxins (P<0.01). PMID- 11064252 TI - Preface. PMID- 11064253 TI - Aspects of the physical chemistry of polymers, biomaterials and mineralised tissues investigated with atomic force microscopy (AFM). AB - Beyond being merely a tool for measuring surface topography, atomic force microscopy (AFM) has made significant contributions to various scientific areas dealing with physical chemistry processes. This paper presents aspects of the physical chemistry at surfaces and interfaces of polymers, biomaterials and tissues investigated with AFM. Selected examples presented include surface induced self-assembly of polymer blends, copolymer interfacial reinforcement of immiscible homopolymers, protein adsorption on biomaterials and erosion of mineralised human tissues. In these areas, AFM is a useful and versatile tool to study structural or dynamic sample properties including thermodynamically driven surface evolution of polymer surfaces, lateral surface composition of interfaces, adsorption processes, and the metrology of demineralisation phenomena. PMID- 11064251 TI - Cultured human mast cells derived from umbilical cord blood cells in the presence of stem cell factor and interleukin-6 cannot be a model of human skin mast cells: fluorescence microscopic analysis of intracellular calcium ion mobilization. AB - To know whether cultured human mast cells raised from umbilical cord blood cells in the presence of stem cell factor (SCF) and interleukin-6 (IL-6) can be a model of human skin mast cells, the cells were stimulated, and intracellular calcium ion ([Ca(2+)](i)) mobilization was analyzed by fluorescence microscopic techniques in parallel with a measurement of histamine released from the cells. When IgE-sensitized mast cells were activated by anti-IgE, [Ca(2+)](i) elevation began at the periphery and subsequently proceeded toward the center of the cells. The increase in [Ca(2+)](i) in calcium ionophore A23187-stimulated mast cells began at the center and spread to the periphery of the cells. Significant histamine release was observed by each stimulation. However, either compound 48/80 or substance P failed to increase [Ca(2+)](i) with no appreciable histamine release. This study shows that there is heterogeneity of [Ca(2+)](i) mobilization in the activated human mast cells, and that cultured human mast cells derived from umbilical cord blood cells in the presence of SCF and IL-6 can not be a model of human skin mast cells. PMID- 11064254 TI - Surface dependent structures of von Willebrand factor observed by AFM under aqueous conditions. AB - von Willebrand factor (VWF) is a large multimeric plasma glycoprotein that adheres rapidly to biomaterial surfaces upon exposure to blood. The adsorbed structure influences subsequent functional interactions with other blood components that mediate surface induced thrombosis. To examine the surface dependent properties of VWF, we compared the adsorbed structures of VWF molecules on two different surfaces: Mica, which is hydrophilic; and octadecyltrichlorosilane (OTS) modified glass, which is hydrophobic. Atomic force microscopy (AFM) was used to image adsorbed VWF under aqueous conditions at physiologic pH and ionic strength. Individual VWF molecules were clearly discernible on both surfaces. On the hydrophobic surface, VWF displayed compact tertiary structures with rare examples of extended molecules. In addition, these data revealed intramolecular structural arrangements of the repeat units within VWF multimers. On the hydrophilic mica surface, VWF displayed extended structures in which intramolecular repeat units were exposed. The lateral dimensions of VWF on mica (640+/-161x303+/-113 nm) were larger than on the hydrophobic OTS (256+/ 74x152+/-62 nm, P<0.005). Our results demonstrate how surface properties mediate the molecular level structure and probable function of VWF, and provide some essential groundwork to develop a mechanistic understanding of surface-induced thrombosis. PMID- 11064255 TI - Changes in the surface structure of purple membrane upon illumination measured by atomic force microscopy. AB - Bacteriorhodopsin (BR) patches with a diameter of 1 to 3 um were investigated in their native state by atomic force microscopy (AFM) in buffer solution. The patches were immobilized deposited and investigated on mica in 150 mM KCl and 10 mM Tris-buffer at pH 8. Under this buffer condition they adsorb preferred with their extracellular side to the solid support mica. The structure of the two dimensional light adapted crystals was resolved with an imaging force of about 100 pN up to a resolution of 13 A. The topography of the surface gets smoother if an imaging force of 1000 pN was applied indicating that protruding structures are compressed. Upon illumination with white light, during imaging with a force of 200 pN, the surface structure of the BR lattice changed. The force- and light induced structural changes were reversible. PMID- 11064256 TI - Atomic force microscopy studies of icosahedral virus crystal growth. AB - Biological macromolecules and particularly viruses, provide excellent systems for the study of crystallization from solution because of their relatively large size. The kinetics of their crystallization is at least an order of magnitude less than for conventional systems, and their large size permits visualization, both of crystal lattices and individual particles, by techniques such as atomic force microscopy (AFM). This technique is especially powerful for biological macromolecules because it can be utilized in situ, in the crystallization mother liquor, over long periods of time without perturbing the growing crystals. We present here observations using AFM of the nucleation and growth of crystals of satellite tobacco mosaic virus, and some recordings as well of bromegrass mosaic virus. Correlations are made, where possible, with corresponding analyses using X ray diffraction analysis. PMID- 11064257 TI - Atomic force microscopy of the three-dimensional crystal of membrane protein, OmpC porin. AB - Three-dimensional microcrystals of OmpC osmoporin were air-dried slowly and imaged in air with an atomic force microscope (AFM). The overall structural features in AFM images are in good agreement with the X-ray diffraction data of these OmpC osmoporin crystals: monoclinic P2(1) with the unit cell constants a=117.6 A, b=110 A, c=298.4 A, beta=97 degrees. Such a good correspondence between X-ray diffraction and AFM data suggests that the slow and mild air-drying of these crystals did not induce any significant alterations in the crystal lattices as expected upon crystal dehydration. At the (100) crystal face, individual trimeric protein-detergent complexes were resolved. These results show the potential for studying the molecular structure of microcrystals of integral membrane proteins. This study also suggests that the crystal grew in a fashion of rapid two-dimensional expansion along the bc plane followed by a slow deposition along the a axis, perhaps as a rate-limiting nucleation process. Thus, AFM imaging of air-dried crystals would also be of considerable use in the early stages of a project to grow large three-dimensional crystals of membrane proteins suitable for high-resolution X-ray diffraction studies. PMID- 11064258 TI - Force measurements on platelet surfaces with high spatial resolution under physiological conditions. AB - Investigations on platelets are essential to understanding the regulation of hemostasis and thrombosis. Activated platelets undergo dramatic conformational and morphological changes mediated by numerous plasma proteins. AFM techniques can combine high spatial resolution with measurements of the mechanical properties of platelet surfaces. Here, we demonstrate two-dimensional force mapping over a human platelet adsorbed on glass under physiological buffer. The best resolution of platelet membrane elasticity we obtained was at 15.6x15.6 nm(2) pixel(-1). In addition, quantitative information on platelet surface charge density was extracted from individual force curves with the aid of DLVO theory. PMID- 11064259 TI - Substrate dependent differences in morphology and elasticity of living osteoblasts investigated by atomic force microscopy. AB - We have used the atomic force microscope (AFM) as a tool for testing the biocompatibility of implant materials by investigating the adhesion behavior of osteoblast cells in vitro. This technique allowed the investigation of cytomorphology and cytomechanical properties of living cells on a submicrometer scale. Cell adhesion was investigated on Cobalt-Chromium (CoCr), Titanium (Ti) and Titanium-Vanadium (TiV) substrates, which are of great interest in the field of implant research. The elastic properties and the morphology of living osteoblasts on the metallic substrates were compared with those of osteoblasts cultured on glass and tissue culture polystyrene (PS). Furthermore, a characterization of the surface roughness of the substrates was performed and the surface coverage of proteins after incubation with cell culture medium on the substrates was observed with the AFM. PMID- 11064260 TI - Microphase separation at the surface of block copolymers, as studied with atomic force microscopy. AB - Atomic force microscopy (AFM) is used to study the phase separation process occurring in block copolymers in the solid state. The simultaneous measurement of the amplitude and the phase of the oscillating cantilever in the tapping mode operation provides the surface topography along with the cartography of the microdomains of different mechanical properties. This technique thus allows to characterize the size and shape of those microdomains and their organization at the surface (e.g. cubic lattice spheres, hexagonal lattice of cylinders, or lamellae). In this study, a series of symmetric triblock copolymers made of a inner elastomeric sequence (poly(butadiene) or poly(alkylacrylate)) and two outer thermoplastic sequences (poly(methylmethacrylate)) is analyzed by AFM in the tapping mode. The microphase separation and their morphology are essential factors for the potential of these materials as a new class of thermoplastic elastomers. Special attention is paid to the control of the surface morphology, as observed by AFM, by the molecular structure of the copolymers (volume ratio of the sequences, molecular weight, length of the alkyl side group) and the experimental conditions used for the sample preparation. The molecular structure of the chains is completely controlled by the synthesis, which relies on the sequential living anionic polymerization of the comonomers. The copolymers are analyzed as solvent-cast films, whose characteristics depend on the solvent used and the annealing conditions. The surface arrangement of the phase-separated elastomeric and thermoplastic microdomains observed on the AFM phase images is discussed on the basis of quantitative information provided by the statistical analysis by Fourier transform and grain size distribution calculations. PMID- 11064261 TI - Tip friction - torsional spring constant determination. AB - A non-destructive technique is presented for verifying torsional spring constants used in lateral force microscopy. Various calibrations of the microscope are required and these are detailed. The technique produces reasonable values which tend to be larger than those predicted from considerations of the cantilever dimensions. The differences are discussed in terms of length corrections and particularly the uncertainty in the thickness of the cantilevers, which has an enormous effect on the values obtained through a priori calculations. Methods for inferring the thickness are discussed. Further, artefacts in conventional force measurements related to the experiments performed here are discussed. PMID- 11064262 TI - Genetic and immunologic characterization of a novel serotype 4, 15 strain of Neisseria meningitidis. AB - The porin proteins of Neisseria meningitidis are important components of outer membrane protein (OMP) vaccines. The class 3 porin gene, porB, of a novel serogroup B, serotype 4, 15 isolate from Chile (Ch501) was found to be VR1-4, VR2 15, VR3-15 and VR4-15 by porB variable region (VR) typing. Rabbit immunization studies using outer membrane vesicles revealed immunodominance of individual PorB (class 3) VR epitopes. The predominant anti-Ch501 PorB response was directed to the VR1 epitope. Anti-PorB VR1 mediated killing was suggested by the bactericidal activity of Ch501 anti-sera against a type 4 strain not expressing PorA or class 5 OMPs. Studies that examine the molecular epidemiology of individual porB VRs, and the immune responses to PorB epitopes, may contribute to the development of broadly protective group B meningococcal vaccines. PMID- 11064263 TI - Super-infection by Bacillus thuringiensis H34 or 3a3b can lead to death in mice infected with the influenza A virus. AB - Bacterial super-infections are the main cause of complication and mortality after influenza virus (IAV) infection. Since Bacillus thuringiensis (Bt) is considered non-pathogenic for humans and is widely sprayed in urban areas, the aim of this work was to evaluate the potential pathogenicity of a combined infection Bt-IAV in a mouse model of pneumonia. Bacteria used for super-infections were Bt serotype H34 isolated from human infection and the insecticidal strain 3a3b obtained from a commercial source. Virus strain was A/Scotland/20/74 (H3N2) adapted to BALB/c mice by serial lung passage. Combined infection with 4% of the viral lethal dose 50% (LD(50)) and 10(2) spores of Bt H34 killed 40% of the mice. Mortality rates increased up to 55% and 100% when combined infections were done with respectively 10(4) and 10(7) spores. The insecticidal strain Bt 3a3b was less pathogenic than Bt H34. A dose of 10(4) spores associated with 4% of IAV LD(50) killed 50% of the mice. This inoculum must be compared with the doses usually sprayed in agriculture: 10(11) spores m(-2). Total protection against super-infection was obtained when mice were treated with amantadine. Even if only a few cases of Bt human infection have been reported, these results suggest a possible risk for workers spraying Bt-based biopesticides during flu outbreaks. PMID- 11064264 TI - Immune-protective antibodies against capsular polysaccharides do not affect natural competence of Streptococcus pneumoniae: implications for current conjugate vaccination strategies? AB - We studied the effect of opsonization of Streptococcus pneumoniae with capsular antibodies on horizontal transfer of DNA. Opsonization did not inhibit DNA uptake. This suggests that horizontal transfer of capsular genes, which is an important escape mechanism of the pathogen, remains a potential threat for the efficacy of conjugate vaccination. PMID- 11064265 TI - Effect of a matrix metalloproteinase inhibitor on host resistance against Listeria monocytogenes infection. AB - Hydroxy acid-based matrix metalloproteinase (MMP) inhibitors have been shown to inhibit tumor infiltration and growth, endotoxin shock, and acute graft-versus host disease. Blockade of the release of soluble tumor necrosis factor-alpha (TNF alpha) and CD95 ligand (CD95L; FasL) from cell-associated forms is reportedly involved in the mechanism of the drug effect. We investigated the effect of a MMP inhibitor, KB-R7785, on host resistance against Listeria monocytogenes infection, in which TNF-alpha is essentially required for the defense, in mice. The administration of KB-R7785 exacerbated listeriosis, while the drug prevented lethal shock induced by lipopolysaccharide and D-galactosamine. KB-R7785 inhibited soluble TNF-alpha production in spleen cell cultures stimulated by heat killed L. monocytogenes and the drug treatment reduced serum TNF-alpha levels in infected mice, whereas the compound was ineffective on the modulation of interferon-gamma and interleukin-10 production. The effect of KB-R7785 was considered to be dependent on TNF-alpha because the drug failed to affect L. monocytogenes infection in anti-TNF-alpha monoclonal antibody-treated mice and TNF-alpha knockout mice. Anti-CD95L monoclonal antibody was also ineffective on the infection. These results suggest that induction of infectious diseases, to which TNF-alpha is critical in host resistance, should be considered in MMP inhibitor-treated hosts. PMID- 11064266 TI - Lipid S, a novel Staphylococcus epidermidis exocellular antigen with potential for the serodiagnosis of infections. AB - We describe the characterisation of a novel glycerophosphoglycolipid (termed lipid S) produced by Staphylococcus epidermidis grown in a chemically defined medium. Lipid S is a short chain length form of the cellular lipoteichoic acid (LTA). It shares common antigenic determinants with LTA, but its chain length of six glycerophosphate units contrasts with 40-42 units in LTA. Lipid S is exocellular and can be recovered from liquid growth medium whereas LTA is associated with the cell wall and membrane. Healthy individuals have low serum levels of IgG against lipid S, but significantly higher titres have been detected in serum from patients with central venous catheter-related sepsis due to coagulase-negative staphylococci and infection of orthopaedic prostheses. An indirect enzyme-linked immunosorbent assay test based on lipid S allows the rapid diagnosis of Gram-positive infection and may have clinical applications in the management of patients with sepsis. PMID- 11064267 TI - Uptake pathways of clinical and healthy animal isolates of Campylobacter jejuni into INT-407 cells. AB - Campylobacter jejuni isolates obtained from human and animal sources showed different invasion levels into human embryonic intestinal (INT-407) cells. There was no significant relation between the degree of invasion and cytotoxins production. The depolymerization of both microfilaments by cytochalasin-D and microtubules by colchicine, demecolcine and nocodazole or stabilization of microtubules by paclitaxel reduced the invasiveness of C. jejuni, although microfilament depolymerization showed greater inhibition than microtubule depolymerization. Interference with receptor-mediated endocytosis by G strophanthin and monodansylcadaverine and inhibition of endosome acidification by monensin reduced the number of viable intracellular C. jejuni cells. Furthermore inhibition of only host protein kinases by staurosporine, but not phosphoinositide 3-kinase by wortmannin or protein kinase-C by calphostin-C, significantly reduced invasion of epithelial cells by C. jejuni. These data suggest that the internalization mechanism triggered by C. jejuni is strikingly different from the microfilament-dependent invasion mechanism exhibited by many of the well-studied enteric bacteria such as enteroinvasive strains of Escherichia coli, Salmonella typhimurium, Shigella flexneri, Yersinia enterocolitica and Yersinia pseudotuberculosis. PMID- 11064268 TI - Identification of a novel antigen from Staphylococcus epidermidis. AB - A genomic DNA library of Staphylococcus epidermidis NCTC 11047 was constructed, using the Lambda Zap Express cloning vector, and screened with serum collected from a patient with S. epidermidis endocarditis. Sequence analysis of a 30 kDa cloned protein, termed staphylococcal secretory antigen, SsaA, identified a novel protein not previously reported in S. epidermidis. SsaA showed strong homology with two other staphylococcal proteins: SceB from Staphylococcus carnosus and a staphyloxanthin biosynthesis protein from Staphylococcus aureus. Further investigation revealed SsaA to be a highly antigenic protein that was expressed in vivo and could be recovered from whole cells and from the culture supernatant. A combination of Western blot analysis and PCR screening identified SsaA or a homologue in 103/103 staphylococcal strains. SsaA-like genes were not detected in other Gram-positive bacteria of medical importance or a number of Gram-negative organisms. Elevated anti-SsaA IgG antibody levels were detected in sera of five patients with S. epidermidis endocarditis but not in patients with other S. epidermidis infections, endocarditis of other aetiologies or patients with no evidence of infection. The expression of SsaA during episodes of S. epidermidis endocarditis suggests a virulence role specific to the pathogenesis of this infectious disease. PMID- 11064269 TI - Application of randomly amplified polymorphic DNA (RAPD) analysis for typing avian Salmonella enterica subsp. enterica. AB - Randomly amplified polymorphic DNA (RAPD) analysis was performed for the molecular genetic typing of 30 Salmonella enterica subsp. enterica strains isolated from chickens and ducks in Thailand. Six different primers were tested for their discriminatory ability. While some of the primers could only differentiate between the different serovars, the use of multiple primers showed that the RAPD method could also subdivide within a given serovar. The Ready-To-Go RAPD analysis beads used, resulted in reproducible and stable banding patterns. As the RAPD technique is simple, rapid and rather cheap, we suggest that it may be a valuable new tool for studying the molecular genetic epidemiology of S. enterica ssp. enterica, both inter- and intra-serovars. PMID- 11064270 TI - The effect of pseudomonas exotoxin A on cytokine production in whole blood exposed to Pseudomonas aeruginosa. AB - To determine the effect of Pseudomonas aeruginosa exotoxin A (P-ExA) on cytokine production, we studied cytokine release induced by heat-killed P. aeruginosa (HKPA) in human whole blood in the presence or absence of P-ExA. P-ExA (0.01-1 microgram ml(-1)) caused a dose-dependent decrease in HKPA-induced production of tumor necrosis factor alpha (TNF), interleukin (IL-) 10, IL-6 and IL-8 (all P<0.05). P-ExA-induced inhibition of IL-10, IL-6 and IL-8 release was not dependent on reduced TNF concentrations, since the relative attenuation of the production of these cytokines was similar in the presence or absence of a neutralizing anti-TNF antibody. The effect of P-ExA on cytokine production may offer a disadvantage to the host with respect to clearance of the infection. PMID- 11064271 TI - Histologic estimation of coronary artery stenoses: reproducibility and the effect of training. AB - Histologic estimation of coronary artery stenoses (CAS) provides the 'gold standard' for clinicopathologic correlations and medicolegal investigations, yet little evidence supports histology as a reproducible diagnostic measure, and none addresses the effect of training on its use. To study these questions, 20 randomly selected Movat-stained coronary artery cross-sections were reviewed 3 times, at 3-month intervals, by six clinical pathologists (CPs), six pathology residents (Res), seven anatomic pathologists (APs), and two cardiovascular pathologists (CVPs). Before the third iteration, a guide to CAS assessment with illustrations was provided. Inter- and intraobserver reproducibility were determined using interclass correlation coefficients (ICC) (0.40-0.75 = fair good; > or = 0.76 = excellent agreement beyond chance). Surprisingly, all study groups had excellent interobserver reproducibility. Before training, at Time 1, the scores were CPs, 0.77; Res, 0.89; APs, 0.93; and CVPs, 0.93. After training, at Time 3, the results were CPs, 0.81; Res, 0.91; APs, 0.86; and CVPs, 0.88. Intraobserver reproducibility for CPs overall was good (ICC, 0.74), and excellent for Res, APs, and CVPs (0.89, 0.94, and 0.97, respectively). In conclusion, statistical analysis failed to demonstrate any significant effect of training or experience on observer reproducibility. PMID- 11064273 TI - Subvalvular left ventricular aneurysms. AB - Subvalvular left ventricular aneurysms can be subaortic or submitral with variable etiology. This is a retrospective study of 19 subvalvular aneurysms seen in 16 cases. There were 12 isolated subaortic aneurysms, 3 isolated submitral aneurysms, and in 1 case with multiple subaortic and submitral aneurysms. Subaortic aneurysms were associated with infective endocarditis, while there seems to be a strong association between submitral aneurysms and tuberculosis. Five cases of subvalvular aneurysm has associated aneurysm sinus of Valsalva-this association suggests a congenital weakness in the attachment of aortic and mitral annuli to the underlying myocardium. Cardio Pathol 2000;9:267-271 PMID- 11064272 TI - Myocardial expression of endothelin-1 in murine Trypanosoma cruzi infection. AB - Chagas' disease, caused by Trypanosoma cruzi, is an important cause of myocarditis and chronic cardiomyopathy and is accompanied by microvascular spasm and myocardial ischemia. We reported previously that infection of cultured endothelial cells with T. cruzi increased the synthesis of biologically active endothlein-1 (ET-1). In the present study, we examined the role of ET-1 in the cardiovascular system of CD1 mice infected with the Brazil strain of T. cruzi and C57BL/6 mice infected with the Tulahuen strain during acute infection. In the myocardium of infected mice myonecrosis and multiple pseudocysts were observed. There was also an intense vasculitis of the aorta, coronary artery, smaller myocardial vessels and the endocardial endothelium. Immunohistochemistry studies employing anti-ET-1 antibody revealed increased expression of ET-1 that was most intense in the endocardial and vascular endothelium. Elevated levels of mRNA for preproET-1, endothelin converting enzyme and ET-1 were observed in the same myocardial samples. Plasma ET-1 levels were significantly elevated in infected CD1 mice 10-15 days post infection. These observations suggest that increased levels of ET-1 are a consequence of the initial invasion of the cardiovascular system and provide a mechanism for infection-associated myocardial dysfunction. PMID- 11064274 TI - Attenuation of isoproterenol-mediated myocardial injury in rat by an inhibitor of polyamine synthesis. AB - OBJECTIVE: Ornithine decarboxylase (ODC) is an initial rate-limiting enzyme in the synthesis of polyamines (putrescine, spermidine, and spermine) that play a role in cell growth and differentiation. Recent studies have shown that spermidine and spermine cause injury to a variety of cells including myocytes in vitro. In this investigation, we used alpha-difluoromethylornithine (DFMO), a specific and irreversible inhibitor of ODC activity and polyamine synthesis to test the hypothesis that polyamines contribute to myocardial injury in rat. METHODS: Male Sprague Dawley rats were treated with (i) saline (0.2 ml/day, s.c.), (ii) isoproterenol (ISO) (5 mg/kg/day for 8 days, s.c.) to produce necrotizing myocardial injury, or with (iii) DFMO + ISO. DFMO was started 2 days before the initiation of ISO and both ISO and DFMO were continued until the end of the experimental period. Myocardial injury was assessed by determining the increased release of creatine phosphokinase (CPK) and lactate dehydrogenase (LDH) into the plasma, and by morphometric analysis of the lesion area in heart sections stained with Gomori trichrome. RESULTS: ISO induced the release of CPK and LDH by 6 hr and 24 hr, respectively, and produced subendocardial necrosis, which was both acute and resolving following 8 days of ISO. DFMO treatment inhibited ISO-induced increases in (i) ODC activity and putrescine and spermidine levels in heart, (ii) CPK and LDH activity in plasma, and (iii) the area of subendocardial lesions. CONCLUSIONS: These observations suggest that polyamines are one of the intracellular factors that contribute to ISO-mediated cardiac injury in the rat. PMID- 11064275 TI - Matrix metalloproteinase expression in nonrheumatic aortic stenosis. AB - BACKGROUND: Nonrheumatic aortic stenosis (NAS) is considered to be a degenerative process characterized by valve thickening, fibrocalcific masses, collagen disarray, and an inflammatory infiltrate. The matrix metalloproteinases (MMPs) are a family of zinc- and calcium-dependent enzymes produced by inflammatory cells that are capable of degrading collagen, elastin, and proteoglycans. This study sought to test the hypothesis that MMPs are involved in the pathogenesis of NAS. METHODS AND RESULTS: Aortic values were obtained from nine patients with NAS undergoing valve replacement and from four patients without NAS during autopsy. Microscopic analysis of NAS specimens revealed variable areas of calcium deposits, fibrosis, and an extensive cellular infiltrate consisting of macrophages, lymphocytes, and fibroblasts. Control aortic valves demonstrated normal architecture, a predominance of fibroblasts, occasional scattered macrophages, and no lymphocytes. Immunohistochemical staining with antibodies to MMP-1, -2, -3 and -9 revealed expression of each enzyme in macrophages, lymphocytes, and fibroblasts of all NAS patients. MMP-1, -2 and -3 were expressed by resident fibroblasts and macrophages in normal valves, but to a lesser degree. MMP-9 was not identified in normal valves. CONCLUSIONS: The current study confirms an inflammatory infiltrate composed of macrophages and lymphocytes in NAS. Additionally, the increased expression of MMP-1, -2, and -3, along with the unique expression of MMP-9 in NAS valve leaflets was documented. These findings are consistent with the hypothesis that NAS is associated with chronic inflammation and that the increased expression of MMPs may contribute to the pathogenesis of this disease process. PMID- 11064276 TI - Giant cell myocarditis as a manifestation of drug hypersensitivity. AB - Adverse drug effects on the myocardium are often classified into toxic and hypersensitivity forms of myocarditis, each with distinct histologic findings. In contrast, giant cell myocarditis (GCM) is generally not associated with adverse drug reactions and has unique histopathologic features. We report four cases of adverse drug reactions in which the histologic findings were characteristic of GCM. The clinical recognition that GCM may be a manifestation of an adverse drug reaction is important, since the prognosis and treatment of this entity may be different from that of other forms of myocarditis. PMID- 11064277 TI - Active infective endocarditis involving all four cardiac valves. AB - A case is presented of a 70-year-old man treated for 3 months for necrotizing pancreatitis with multiorgan failure. The autopsy revealed enterococcal endocarditis affecting all eleven valvular cusps of the four heart valves. PMID- 11064278 TI - Coronary arteritis with marked fibrous periarteritis: case report. AB - A case is presented of a 43-year-old man who died suddenly and unexpectedly from cardiac malignant arrhythmia. Autopsy revealed isolated coronary arteritis involving primarily the left coronary stem and extending into the proximal half of its anterior descending branch. The most striking feature of the disease was marked fibrous periarteritis. Similar changes were present segmentally in the peripheral third of the right coronary artery, and also as small isolated foci of adventitial fibrosis in the aortic base. With some hesitation, we classify the case as coronary Takayasu's arteritis. PMID- 11064279 TI - The role of the antennal glands in ion and body volume regulation of cannulated Penaeus monodon reared in various salinity conditions. AB - Urinary production rate and the osmotic and ionic concentrations in both urine and hemolymph were measured in cannulated intermolt Penaeus monodon which were either abruptly transferred from 45 ppt seawater to 15 ppt seawater (Experiment 1) or acclimated to 5, 25 and 45 ppt seawater (Experiment 2). In Experiment 1, urinary magnesium concentration fell dramatically from 228 to 30 mEq/l within 4 h post-transfer, but 8 h after transfer, U/H (urine/hemolymph) ratios stabilized at between 1.0 and 2.5. Sodium was higher in urine than in hemolymph during the first 24 h after transfer, while potassium was lower in urine than in hemolymph until 72 h after transfer, which suggests that sodium and potassium concentrations are regulated by the antennal gland after an abrupt change in media. In Experiment 2, the urinary production rate of P. monodon decreased as salinity increased, suggesting that the antennal glands also regulate body volume. In the acclimated shrimps of Experiment 2, the antennal glands did not appear to regulate osmolarity or the concentration of chloride, sodium, potassium, and calcium ions, but as salinity increased, U/H ratios of magnesium increased from 2.3 to 13.5, and active secretion by the antennal gland accounted for 57 approximately 93% of the total magnesium excretion through urine. These results suggest that active secretion of magnesium by the antennal gland enable this shrimp to maintain hypoionic levels of magnesium in the hemolymph. PMID- 11064280 TI - Resting metabolism and heat increment of feeding in mandarin fish (Siniperca chuatsi) and Chinese snakehead (Channa argus). AB - Resting metabolism was measured in immature mandarin fish Siniperca chuatsi weighing 42.1-510.2 g and Chinese snakehead Channa argus weighing 41.5-510.3 g at 10, 15, 20, 25, 30 and 35 degrees C. Heat increment of feeding was measured in mandarin fish weighing 202.0 (+/-14.0) g and snakehead weighing 200.8 (+/-19.3) g fed swamp loach Misgurnus anguillicaudatus at 1% body weight per day at 28 degrees C. In both species, weight exponent in the power relationship between resting metabolism and body weight was not affected by temperature. The relationship between resting metabolism and temperature could be described by a power function. The temperature exponent was 1.39 in mandarin fish and 2.10 in snakehead (P<0.05), indicating that resting metabolism in snakehead increased with temperature at a faster rate than in mandarin fish. Multiple regression models were used to describe the effects of body weight (W, g) and temperature (T, degrees C) on the resting metabolism (R(s), mg O(2)/h): lnR(s)=-5.343+0.772 lnW+1.387 lnT for the mandarin fish and lnR(s)=-7.863+0.801 lnW+2.104 lnT for the Chinese snakehead. The proportion of food energy channelled to heat increment was 8.7% in mandarin fish and 6.8% in snakehead. PMID- 11064281 TI - Effects of animal or plant protein diets on cecal fermentation in guinea pigs (Cavia porcellus), rats (Rattus norvegicus) and chicks (Gallus gallus domesticus). AB - Monogastric herbivores such as the guinea pig depend on energy supply from enteric fermentation as short-chain fatty acids (SCFA) corresponding to 30-40% of their maintenance energy requirements. They evolved specific digestive system to adapt their indigenous microflora to plant polysaccharides fermentation. No information has been available about the adaptability of microbial fermentation in hindgut of the monogastric herbivorous to an animal protein diet. We investigated if the guinea pig can fully retrieve energy of an animal protein diet by hindgut fermentation compared with a plant protein diet. For comparison, we also studied two omnivores. End products of in vitro cecal fermentation (SCFA, ammonia and gases) were measured to judge how well an animal protein diet could be fermented. The animal protein diet resulted in the less intensive fermentation with increased feed intake and volume of cecal contents than the plant protein diet only in guinea pigs. This may be due to a limited capacity of the hindgut microflora to adapt to the substrate rich in animal protein. We also found that chick cecal contents produced methane at higher emission rate than ruminants. PMID- 11064282 TI - Temperature adaptation influences the aggregation state of hemocyanin from Astacus leptodactylus. AB - When Astacus leptodactylus were kept at various temperatures for several weeks, different ratios between di-hexameric and hexameric hemocyanins were observed in their hemolymph. The higher the temperature the more hexamers were present. This long-term adaptation to different temperatures or/and to temperature-induced pH shifts as observed in the hemolymph has different effects on the expression of subunit types building up hexamers and those which covalently link two hexamers within the di-hexamers. The oxygen binding behaviour of di-hexameric hemocyanins from cold and warm adapted animals do not show differences with respect to affinity, Bohr effect and cooperativity. PMID- 11064283 TI - Amino acids in haemolymph, single fibres and whole muscle from the claw of freshwater crayfish acclimated to different osmotic environments. AB - The concentrations of free amino acids were measured in whole claw muscle, single fibres and haemolymph of Australian freshwater crayfish, Cherax destructor, during the intermoult stage. The average total pool of amino acids in short sarcomere fibres (179 mmol kg(-1)) was 60% greater than in long-sarcomere fibres, due to higher concentrations of alanine, cysteine, glutamate, leucine and proline. The two fibre types exhibited differences in the banding pattern of the isoforms of troponin using gel electrophoresis. The average pool of amino acids in haemolymph was 2.7 mmol kg(-1). Cherax has symmetrical claws and the total pool of amino acids from whole muscles (approx. 79 mmol kg(-1)) was similar in left and right claw muscles. In animals acclimated to osmotic environments between 0 and 220 mOsm, the osmotic pressure of the haemolymph increased from 356 to 496 mOsm, but no systematic changes were observed in the amino acid profiles of muscles or haemolymph. The major findings were that (a) concentrations of amino acids differed between the two major fibre types in claw muscle and (b) amino acids in the muscle fibres did not play a major part in intracellular osmoregulation in Cherax, suggesting this species is an anisosmotic regulator. PMID- 11064284 TI - Control of melanosome movement in intact and cultured melanophores in the bitterling, Acheilognathus lanceolatus. AB - The melanophores in the dermis on scales in the bitterling, Acheilognathus lanceolatus were studies to obtain information about the control mechanism of aggregation and dispersion using intact, membrane-permeabilized and cultured cells. The cultured melanophores showed supersensitivity, namely, they responded to norepinephrine with much higher sensitivity than intact cells. The cultured melanophores failed to respond to high KCl. Melatonin aggregated and adenosine dispersed melanosomes within a cell. Digitonin permeabilized cells showed aggregation with Ca ions and dispersion by cyclic adenosine 3',5'-monophosphate (cAMP) in the presence of ATP. Movement of melanosomes was observed under the high magnification of light microscope and the tracks of each pigment granule were followed. The granules moved fast and linearly during aggregation, whereas they showed to-and-fro movement during dispersion. PMID- 11064285 TI - Characterization of parameters for in vitro culture of isolated ovarian follicles of greenback flounder Rhombosolea tapirina. AB - Isolated ovarian follicles of greenback flounder Rhombosolea tapirina were incubated with a variety of gonadotropins (GtHs) and steroid precursors for periods of up to 42 h, and levels of free and glucuronated testosterone (T) and 17beta-estradiol (E(2)) in the medium, and free T and E(2) from inside follicles were measured by RIA. Short incubations (6 h) generated increases in T and E(2) in response to steroid precursors, but not human chorionic GtH (hCG), or salmon or carp GtH. At incubation times of 18 h, all GtHs stimulated T and, or E(2) production, whereas after 42-h incubation, GtH effects on E(2) production had disappeared. Steroid precursors remained effective at 18 and 42 h. T and E(2) glucuronides were formed in small quantities but did not account for loss of treatment effects at long incubation times. Instead, this could be explained by accumulation of E(2) in controls as a result of continued basal steroid production. Follicles absorbed substantial amounts of both endogenous and exogenous steroid from the medium, however, this did not appear to have any influence on changes in treatment effects with incubation time. Flounder follicles were most sensitive to hCG, followed by salmon and carp GtH at approximately 10-fold higher concentrations. Ovarian segments were not sensitive to any GtH but did convert exogenous steroid precursors indicating that tissue access by GtH may be a limiting factor under certain in vitro conditions. HCG augmented the conversion of 17-hydroxyprogesterone (17P) to T but not T to E(2), consistent with the relative GtH-insensitivity of aromatase in other species. Follicles converted a range of steroid precursors with equal competence, indicating that no step in the cleavage pathway is strongly rate-limited, and that choice of precursor is unlikely to affect the assessment of steroidogenic activity. PMID- 11064286 TI - Bovine interleukin-1 expression by cultured mammary epithelial cells (MAC-T) and its involvement in the release of MAC-T derived interleukin-8. AB - MAC-T cells, an established bovine mammary epithelial cell line, were utilized to investigate both expression of interleukin-1 (IL-1) mRNA and secretion of IL-1 after Escherichia coli lipopolysaccharide (E. coli LPS) stimulation. In addition, recombinant human IL-1beta, recombinant human IL-1 receptor antagonist (IL-1ra) and a neutralizing goat antibody against type I human IL-1 receptor were used to study the involvement of IL-1 in the release of IL-8. The expression of MAC-T derived IL-1alpha mRNA was correlated to production of IL-1alpha protein as measured by an IL-1alpha sandwich ELISA. Secretion of IL-1alpha was dose- and time-dependent, with a maximal level of 600 pg/ml detectable upon 2-h stimulation with 20 microg of LPS per ml. IL-1ra and the neutralizing antibody significantly blocked the ability of IL-1beta to stimulate secretion of IL-8 by MAC-T cells. During this study, we have demonstrated that MAC-T cells secrete IL-1 in response to LPS stimulation and IL-1 is an important mediator for the release of the bovine IL-8 by MAC-T cells. These results further indicate the potential importance of mammary epithelial cells as a source of immunoregulation in the mammary gland via cytokine elaboration. PMID- 11064287 TI - Respiration in the burrowing brittlestar, Hemipholis elongata say (Echinodermata, Ophiuroidea): a study of the effects of environmental variables on oxygen uptake. AB - The burrowing brittlestar Hemipholis elongata (Say) maintains a constant M(O2)of 3.79+/-1.47 micromol O(2) g(-1) h(-1) (for 0.2-0.3 g animals, mean+/-S.D., n=7), measured in the burrow, over a broad range of PO(2). Below the critical PO(2) of 37 mm Hg, M(O(2)) becomes dependent on the oxygen tension. M(O2) is a function of the size of H. elongata; the scaling exponent is 0. 83 and is similar to those reported for other echinoderms. The M(O2) of H. elongata is unaffected by removal from the burrow, by hypercapnia, by exposure to hydrogen sulfide, or by temperature change in the range from 20 to 32 degrees C. The relative insensitivity of H. elongata to these factors may be an adaptation to life in the highly variable estuarine and tidal creek environments where the animals are frequently found. PMID- 11064288 TI - Development of an anti-vitellin ELISA for the assessment of reproduction in the ridgeback shrimp, Sicyonia ingentis. AB - To investigate the reproductive regulation of the ridgeback shrimp, Sicyonia ingentis, vitellin (Vn) synthesis was studied. Using gel filtration chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Vn was found to have a molecular mass of 322 kDa and to be composed of three subunits of 182, 91 and 85 kDa. Purified Vn was used to prepare anti-Vn antiserum, which was used to develop an enzyme-linked immunosorbent assay (ELISA) with a dynamic range of 0.3-300 ng. The ELISA was used to measure hemolymph levels of yolk proteins. The mean hemolymph concentrations in fresh caught animals ranged from 256 (+/-36.6 S.E.M.) to 1073 (+/-87.6 S.E.M.) mg/ml in stage 2 and 4 animals, respectively. The ELISA was also used to determine the effects of steroid hormone injections in adult non-reproductive female shrimp. One milligram injections of progesterone, 17alpha-hydroxyprogesterone or 17beta estradiol were administered for three consecutive days to individual females. There were no changes in hemolymph vitellogenin levels during the successive 7 day period following the first injection of any steroid. PMID- 11064289 TI - Milk intake in blue fox (Alopex lagopus) and silver fox (Vulpes vulpes) cubs in the early suckling period. AB - Milk intake of fox cubs (2-16 days of age; body weight, 96-649 g) in ten blue fox litters and ten silver fox litters were measured by the water isotope dilution (WID) technique following a single intraperitoneal injection of tritiated water (3HHO). Litter size varied from four to 14 in blue foxes and from three to eight in silver foxes. Silver fox cubs had higher birth weights than blue foxes. Inter species body weights and growth rates were apparently dependent on litter size and the dam's constitution. In both species growth rate increased with age and body weight (7-35 g per day). In the cubs, the biological half-life of body water turnover (BWT) rose from 1.5 days at 2-3 days of age to 2.5 days at 13-16 days of age, although a considerable scatter was seen. The mean daily milk intake of the cubs varied with body weight, from 31 to 193 g per day, whereas daily milk intake per unit of body mass remained stable at 30-35 g per 100 g body weight. The ratio of milk intake to body weight gain varied considerably among cubs, averaging 4.5 g/g during the 3-week experimental period. In suckling fox cubs, the calculated daily intake of metabolically energy (ME) corresponded fairly with the estimated energy requirements for growth and maintenance of the young. Finally, the applicability and the accuracy of the WID technique was evaluated in ten 3-week old fox cubs, by tube-feeding with a milk replacer for 48 h, which documented that the daily rates of milk intake and water turnover can be accurately measured in suckling fox cubs by the WID technique following a single injection of 3HHO. PMID- 11064290 TI - Host defense function in neutrophils from the American bison (Bison bison). AB - Selected host defense functions of neutrophils isolated from American bison (Bison bison) were characterized and compared with those of cattle (Bos taurus). Bison neutrophils had a robust chemotactic response to both IL-8 and LTB(4), with maximal responses occurring at 10(-7) M (IL-8) and 10(-8) M (LTB(4)). The magnitude of the chemotactic response to IL-8 was similar in bison and bovine neutrophils (except at 10(-7) M IL-8, where bison had a stronger response). In response to LTB(4), bison neutrophils had a much stronger chemotaxis at both 10( 8) and 10(-7) M than did bovine cells. Production of reactive oxygen species (ROS) in response to phorbol myristate acetate (PMA) and opsonized zymosan (OpZ) was similar between bison and bovine neutrophils. However, the production of ROS in bison neutrophils stimulated with OpZ was primarily intracellular, while extracellular release of ROS was evident in bovine neutrophils stimulated with OpZ. Like bovine neutrophils, bison neutrophils did not generate a respiratory burst in response to fMLF. Granules prepared from bison neutrophils had potent direct killing action on the Gram-negative bacteria Escherichia coli but failed to kill the Gram-positive bacteria Staphylococcus aureus and, at intermediate doses, actually had a permissive effect for this bacteria. Thus, bison neutrophils have potent host defense capabilities similar in quality to those of bovine neutrophils; however, unique differences are present, which may allow bison neutrophils to respond to the distinct immunological challenges that bison encounter. PMID- 11064291 TI - Sustainability of Canadian medicare: optimism for the future (Part II). PMID- 11064292 TI - The International Guidelines 2000 for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care. Heart and Stroke Foundation of Canada. PMID- 11064293 TI - Cyberwires. PMID- 11064294 TI - Country mouse/city mouse: differences in Canadian and American care of the ST elevation myocardial infarction patient. PMID- 11064295 TI - Overview of 'APPROACH' - the Alberta Provincial Program for Outcome Assessment in Coronary Heart Disease. PMID- 11064296 TI - Overview of the Alberta Provincial Project for Outcome Assessment in Coronary Heart Disease. On behalf of the APPROACH investigators. AB - The Alberta Provincial Project for Outcome Assessment in Coronary Heart Disease (APPROACH) is an ongoing prospective data collection initiative that began in January 1995. The cohort for the initiative is all patients undergoing cardiac catheterization in Alberta. Patients are followed longitudinally for the determination of short and long term clinical, economic and quality of life outcomes. The project is producing valuable information on the processes and outcomes of cardiac care in Alberta, and is now being implemented in British Columbia as well. This paper provides an overview of APPROACH with specific attention to the project's general objectives, salient features, database structure and technical specifications. Examples of applied research projects based on APPROACH data are also provided. PMID- 11064297 TI - Temporal evolution in the management of acute ST elevation myocardial infarction: the seven-year GUSTO experience from canada and the united states. The North American GUSTO-I and GUSTO-III investigators. AB - BACKGROUND: Temporal changes in baseline characteristics, treatment and clinical outcomes of patients presenting with acute ST elevation myocardial infarction in Canada and the United States have not been examined comprehensively over time. OBJECTIVES: To evaluate baseline characteristics, process of care and clinical outcomes. Also, to explore whether earlier process-of-care differences between Canada and the United States had changed and, if so, whether they influenced clinical outcomes. PATIENTS AND METHODS: A total of 13, 888 American and 3011 Canadian patients enrolled in 184 American and 38 Canadian hospitals that participated in both the Global Utilization of Streptokinase and Tissue Plasminogen Activator (alteplase) for Occluded Coronary Arteries (GUSTO-I) trial (1990 to 1993) and the Global Utilization of Strategies to Open Occluded Coronary Arteries (GUSTO-III) trial (1995 to 1997) were studied. Logistic regression was used to identify significant prognostic factors, to assess illness severity at hospital presentation and to classify trends between Canada and the United States. RESULTS: In both countries, illness severity on admission increased, door to-needle time for thrombolysis was reduced, intensive care unit stay was shortened and hospital stay decreased from GUSTO-I to GUSTO-III. Whereas the administration of oral nitrates, calcium blockers and beta-blockers at discharge converged over time between countries, the disparity in the use of angiography and revascularization widened; the rise in American revascularization rate was most evident in patients without in-hospital ischemia. The 30-day and one-year mortality rates were comparable and declined nonsignificantly in both countries. CONCLUSIONS: Despite increased illness severity and varying medication and procedure rates, there was no increase over time in 30-day or one-year mortality; this remained comparable between countries throughout the seven-year observation period. PMID- 11064298 TI - The crochetage pattern in electrocardiograms of pediatric atrial septal defect patients. AB - OBJECTIVES: To test the sensitivity and specificity of the crochetage pattern (a notch near the apex of the R wave in electrocardiographic inferior limb leads) in the pediatric electrocardiogram for detecting patients with a secundum atrial septal defect. PATIENTS AND METHODS: Electrocardiograms from 82 consecutive preoperative pediatric patients with a secundum atrial septal defect confirmed by two-dimensional echocardiography were reviewed for evidence of right ventricular hypertrophy and the crochetage pattern. These electrocardiograms were compared with 244 consecutive preoperative controls consisting of patients with echocardiographically proven patent foramen ovale, ventricular septal defect, pulmonary stenosis, tetralogy of Fallot and patients with normal echocardiogram studies. RESULTS: The electrocardiographic crochetage pattern was observed in 31.7% of preoperative patients with a secundum atrial septal defect in at least one inferior limb lead. The specificity of the crochetage pattern for the detection of a secundum atrial septal defect was high when present in all three inferior limb leads (greater than 92%). The crochetage pattern in at least one lead in secundum atrial septal defects shows no association with incomplete right bundle branch block (c2(1)=0.80, not significant), and thus these two findings together do not improve the detection of an atrial septal defect. CONCLUSIONS: The sensitivity and specificity of the electrocardiographic crochetage patterns in at least one inferior limb lead in echocardiographically proven secundum atrial septal defects are 31.7% and 86.1%, respectively. The electrocardiographic crochetage or notching pattern in inferior limb leads has a high specificity for atrial septal defects in the pediatric population. PMID- 11064299 TI - Levels of evidence in cardiovascular clinical practice guidelines. AB - BACKGROUND: Clinical practice guidelines (CPGs) can be helpful in distilling the medical research literature for clinicians; however, the guidelines should acknowledge the variable methodological quality used in clinical research by tempering their recommendations with a 'levels of evidence' scale. OBJECTIVE: To evaluate the proportion of English-language cardiovascular CPGs that provide the user with recommendations graded according to a defined levels of evidence scale. In addition, to evaluate other key aspects important in the critical appraisal of CPGs. METHODS: CPGs for atrial fibrillation, congestive heart failure and myocardial infarction were identified by searching MEDLINE, a reference text of CPGs and the Internet. Each CPG was evaluated using a priori-defined criteria based on the Evidence-Based Medicine Working Group's paper on critical appraisal of CPGs, including use of a reproducible search strategy, method of obtaining consensus, peer review and testing in practice. RESULTS: A total of 95 CPGs were evaluated. Only 13% graded their recommendations using a defined levels of evidence scale. In addition, few CPGs documented a reproducible search strategy or peer review process, and none had been formally tested in practice. CONCLUSIONS: Reporting the levels of evidence for recommendations is an important component of CPGs, yet this system is not widely used. PMID- 11064300 TI - Impact of a mandatory physician reporting system for cardiac patients potentially unfit to drive. AB - CONTEXT: Sudden cardiac incapacitation of a driver may lead to the death or serious injury of passengers or bystanders. This has raised public safety concerns and has led to the creation of legislation to protect the public. Some jurisdictions in Canada and the United States have introduced mandatory physician reporting of patients who may be unfit to drive for medical reasons. The impact on motor vehicle accident (MVA)-related morbidity and mortality of mandatory physician reporting for at-risk cardiac patients is unknown. OBJECTIVE: To determine the impact of mandatory physician reporting legislation (for cardiac patients) in Ontario (population 10.3 million) on MVA-related morbidity and mortality. DATA SOURCES: Reporting data were obtained from the Ontario Ministry of Transportation. Incidence and prevalence data were taken from Ontario Ministry of Health sources and from the literature (MEDLINE). Data for modelling were taken from the literature (MEDLINE) and from the Canadian Cardiovascular Society's Consensus Conference document on cardiac illness and fitness to drive. DATA EXTRACTION: Licence suspension data (correlated with medical illness) were taken directly from government documents. These were then applied to a 'risk of harm' formula used to calculate the risk posed to bystanders and passengers by the suspended patients if they had continued to drive. Canadian licence suspension guidelines were then reviewed in conjunction with cardiac disease incidence and prevalence data to arrive at the number of patients who should have been suspended. Physician compliance with the legislation was then calculated, along with the potential impact on MVA-related morbidity and mortality in the scenario of 100% physician compliance. STUDY SELECTION: All Ontario drivers who had licence suspensions in 1996 for reasons of cardiac disease were included in the analysis. DATA SYNTHESIS: Nine hundred and ninety-four licences were suspended for cardiac reasons in 1996, compared with an estimated 72,407 that should have been suspended if Canadian guidelines had been followed (1.4%). Less than one death or serious injury was avoided as a result of the legislation (from the 'risk of harm' formula). If all drivers with cardiac illness had been suspended from driving, up to 29.2 such events could potentially have been avoided. However, only 13 of 929 (1.4%) road fatalities in Ontario in 1996 were attributed to a driver with a medical illness. CONCLUSIONS: Mandatory physician reporting of patients with cardiac illness has a negligible impact on MVA-related morbidity and mortality. PMID- 11064301 TI - Duromedics original prosthesis: what do we really know about diagnosis and mechanism of leaflet escape? AB - The Edwards-Duromedics (ED) is a bileaflet pyrolitic carbon mechanical valve introduced by Hemex Scientific Inc in 1982, subsequently acquired by Baxter Healthcare Corp, withdrawn from the market in 1988, and modified and reintroduced in 1990. From 1982 to date, 46 cases of leaflet escape have been registered by the manufacturer of an estimated total of 20,000 valves implanted. Disc embolization 12 years after an ED mitral prosthesis implantation is reported in a 45-year-old man operated on when he was in cardiogenic shock because a preliminary transthoracic Doppler echocardiography did not show malfunction of the valve. A correct diagnosis was made four days after the onset of the symptoms by transesophageal echocardiography. During the operation, the posterior leaflet of the ED valve was not found, a 29 mm St Jude Medical bileaflet mechanical prosthesis was implanted and the patient died in the intensive care unit because of low cardiac output syndrome. Cavitation damage is generally considered the most frequent mechanism in cases of such fracture. Thus, any patient with a mechanical valve presenting with acute pulmonary edema must be immediately transferred to a surgical unit; cinefluoroscopy or transesophageal echocardiography may be performed rapidly to achieve successful management of patients with leaflet embolization. PMID- 11064302 TI - Transesophageal echocardiography for detection of mitral regurgitation due to papillary muscle rupture or dysfunction associated with acute myocardial infarction: a report of five cases. AB - Severe mitral regurgitation was associated with cardiogenic shock in five (0.8%) of 623 patients with acute myocardial infarction who were urgently admitted to the authors' hospitals between 1994 and 1996. The infarct was located in the inferior wall in four patients and in the inferoposterior wall in one patient. Severe mitral valve regurgitation occurred concurrently with cardiogenic shock between one and six days after the onset of myocardial infarction. A mitral regurgitant murmur was not audible in four of five patients. Similarly, mitral regurgitant Doppler signals were not detected in four patients by transthoracic echocardiographic examination, while transesophageal echocardiographic examination detected mitral regurgitant signals clearly in all patients. Thus, when cardiogenic shock is unexpectedly associated with inferior or inferoposterior wall acute myocardial infarction, severe mitral regurgitation should be suspected, even when a mitral regurgitant murmur is not audible. Furthermore, mitral regurgitant flow signals may not always be detected by transthoracic echocardiography. Thus, examination for mitral regurgitation by transesophageal echocardiography should be considered. PMID- 11064303 TI - Isolated pulmonic valve endocarditis in healthy hearts: a case report and review of the literature. AB - The case of a 53-year-old man with isolated pulmonic valve endocarditis in a structurally normal heart is presented. The patient had a history of chronic obstructive pulmonary disease and was admitted to hospital with an apparent exacerbation with pneumonia. Blood cultures grew Staphylococcus aureus, and an echocardiogram identified a large vegetation on the pulmonic valve in a structurally normal heart. He was unsuccessfully treated with antibiotics and eventually required pulmonic valve replacement. The literature from 1960 to 1999 identified only 36 reported cases of pulmonic valve endocarditis in structurally normal hearts. The present report underscores the importance of suspecting pulmonic valve endocarditis in patients with multiple pulmonary lesions, and discusses the predisposing factors, clinical features, diagnostic role of echocardiography and the potential benefits of early surgical treatment. PMID- 11064304 TI - Chapter 1. Summary of the CCS Consensus Conference on prevention of sudden death from cardiac arrhythmia. PMID- 11064305 TI - Chapter 2. Guidelines and recommendations for ICD therapy. PMID- 11064306 TI - Chapter 3. Cost effectiveness of ICD therapy: a review of published evidence. PMID- 11064307 TI - Chapter 4. The ethics of setting limits on ICD therapy. PMID- 11064308 TI - Chapter 5. Legal implications of ICD therapy. PMID- 11064309 TI - Implantable cardioverter/defibrillator therapy: the meaning of terms. PMID- 11064310 TI - Expression of Lewis(a), sialyl Lewis(a), Lewis(x) and sialyl Lewis(x) antigens as prognostic factors in patients with colorectal cancer. AB - BACKGROUND: Altered expression of blood group-related carbohydrate antigens such as sialyl Lewis (Le)(x) antigen in tumours is associated with tumour progression behaviour and subsequent prognosis. However, the prognostic value of the expression of Le-related antigens in colorectal tumours remains unclear. PURPOSE: To clarify the prognostic value of Le(a), sialyl Le(a), Le(x) and sialyl Le(x) expression in colorectal carcinomas as prognostic factors after surgery. PATIENTS AND METHODS: Colorectal carcinoma samples from 101 patients with primary colorectal carcinoma who underwent surgical resection were subject to immunohistochemical analyses for Lea, sialyl Lea, Lex and sialyl Le(x) expression with the respective monoclonal antibodies. RESULTS: Le(a), sialyl Le(a), Le(x) and sialyl Le(x) were expressed in 69 (68.3%), 73 (72.3%), 66 (65.4%) and 76 (75.3%) carcinomas, respectively. The patients with sialyl Lex-expressing tumours had more advanced cancer than those with nonsialyl Lex-expressing tumours (P=0.0029). The survival time after surgery of patients with Le(x)- or sialyl Le(x)-expressing tumours was significantly shorter than the survival time of those with non-Le(x)- or nonsialyl Le(x)-expressing tumours, respectively (P=0.023 and P=0. 0001, respectively). Cox's regression analysis revealed that Le(x) and sialyl Le(x) expression, separate from stage and histological type, were prognostic variables for patient survival (hazard ratio [HR] for sialyl Le(x)-positive expression to sialyl L(x)-negative expression 2.90; HR for Le(x) positive expression to Le(x)-negative expression 12.76 in stage I/IV, 0.63 in stage II and 1.69 in stage III). CONCLUSIONS: Le(x) expression and sialyl Le(x) expression in colorectal carcinomas are each associated with poor prognosis. These variables should be considered in the design of future trials. PMID- 11064311 TI - Does the depth of gastric ulceration influence a modified dual therapy with amoxicillin and lansoprazole for Helicobacter pylori-associated gastric ulcer? AB - PURPOSE: To clarify whether the depth of ulceration evaluated by endoscopic ultrasonography (EUS) influences a modified dual therapy with amoxicillin and lansoprazole for the treatment of Helicobacter pylori-positive patients with gastric ulcer. PATIENTS AND METHODS: Twenty-two consecutive cases of gastric ulcer (nine superficial ulcers and 13 deep ulcers) in H pylori-positive patients were studied. Ten of 22 patients received a two-week eradication therapy with amoxicillin 1500 mg/day, lansoprazole 30 mg/day and a new antiulcer agent with features in common with sucralfate, ecabet sodium, 2.0 g/day. They continued to receive the same doses of lansoprazole and ecabet sodium for the next six weeks. The other 12 patients received the same therapy except for those who underwent the four-week amoxicillin treatment. All patients underwent EUS both at the start of the study and eight weeks later. They then received ecabet sodium alone for the next six months as a maintenance therapy, followed by a six-month interval with no treatment. The final endoscopy was done one year after H pylori eradication therapy was completed to evaluate H pylori status and ulcer recurrence. RESULTS: The rates of endoscopic healing and H pylori eradication in the nine patients with superficial ulcer were 100%, irrespective of the period of amoxicillin treatment. In contrast, the rates of endoscopic evidence of healing and H pylori eradication in the 13 patients with deep ulcer were different for each period of amoxicillin treatment; that is, the rates of reduction in ulcer determined by echo and H pylori eradication in the four patients treated with the two-week amoxicillin course were significantly lower (P=0.03) than those in the nine patients treated with the four-week course. CONCLUSION: Ulcer depth is likely to influence the success of amoxicillin treatment for H pylori-positive patients with gastric ulcer. PMID- 11064312 TI - Health information provided by retail health food outlets. AB - Alternative health practices have become increasingly popular in recent years. Many patients visit specific complementary practitioners, while others attempt to educate themselves, trusting advice from employees at local health food stores or the Internet. Thirty-two retail health food stores were surveyed on the nature of the information provided by their staff. A research assistant visited the stores and presented as the mother of a child in whom Crohn's disease had been diagnosed. Seventy-two per cent (23 of 32) of store employees offered advice, such as to take nutritional and herbal supplements. Of the 23 stores where recommendations were made, 15 (65%) based their recommendation on a source of information. Fourteen of the 15 stores using information sources used the same reference book. This had a significant impact on the recommendations; the use of nutritional supplements was favoured. In conclusion, retail health food stores are not as inconsistent as hypothesized, although there are many variances in the types of supplements recommended for the same chronic disease. PMID- 11064313 TI - Intrafamilial clustering of Helicobacter pylori infection in Saudi Arabia. AB - AIM: To study the pattern of Helicobacter pylori infection among family members in the Saudi population. METHODS: A cross-sectional, population-based, seroepidemiological study of family members was undertaken in a Saudi population using saliva H pylori immunoglobulin (Ig) G antibodies (Helisal kit). RESULTS: A total of 42 families comprising 271 children and 84 parents were studied (355 subjects; mean age 23 years, SD 19 years) The overall frequencies of H pylori IgG antibodies in mothers, fathers and children were 67%, 64% and 23%, respectively. There was no significant difference in the infection rate between mothers and fathers, or between boys and girls. The infection rate among children increased when one or both parents were seropositive, and the infection rate among parents was proportionally related to the number of infected children per family. The frequency of H pylori antibodies was significantly higher in spouses of seropositive parents than in spouses of seronegative parents (45% compared with 19.2%). CONCLUSIONS: These data confirm that the intrafamilial clustering of H pylori infection in Saudi Arabia occurs in a similar pattern to that described in the developed countries, and that living conditions and social conditions lead to person to person transmission of H pylori infection. PMID- 11064314 TI - Indications for liver transplantation in British Columbia's Aboriginal population: a 10-year retrospective analysis. AB - OBJECTIVES: To study the indications for liver transplantation among British Columbia's First Nation population. MATERIALS AND METHODS: A retrospective analysis of the British Columbia Transplant Society's database of Aboriginal and non-Aboriginal liver transplant recipients from 1989 to 1998 was undertaken. For primary biliary cirrhosis (PBC), the transplant assessment database (patients with and without transplants) was analyzed using a binomial distribution and compared with published census data regarding British Columbia's proportion of Aboriginal people. RESULTS: Between 1989 and 1998, 203 transplantations were performed in 189 recipients. Fifteen recipients were Aboriginal (n=15; 7.9%). Among all recipients, the four most frequent indications for liver transplantation were hepatitis C virus (HCV) infection (n=57; 30.2%), PBC (n=34; 18.0%), alcohol (n=22; 11.6%) and autoimmune hepatitis (n=14; 7.4%). Indications for liver transplantation among Aboriginal people were PBC (n=8; 53.3%; P<0.001 compared with non-Aboriginal people), autoimmune hepatitis (n=4; 26.67%; P=0.017), acute failure (n=2; 13.3%) and HCV (n=1). Among all patients referred for liver transplantation with PBC (n=43), 29 (67.44%) were white and 11 (25.6%) were Aboriginal. A significant difference was found between the proportion of Aboriginal people referred for liver transplantation and the proportion of Aboriginal people in British Columbia (139,655 of 3,698,755 [3.8%]; 1996 Census, Statistics Canada) (P<0.001). CONCLUSIONS: Aboriginal people in British Columbia are more likely to be referred for liver transplantation with a diagnosis of PBC but are less likely to receive a liver transplant because of HCV or alcohol than are non-Aboriginal people. PMID- 11064315 TI - Transgenic technology and the study of hepatitis viruses: a review of what we have learned. AB - Because of the absence of inbred animal models susceptible to infection by the hepatitis B (HBV), C (HCV) and delta (HDV) viruses, and the inability to culture these viruses, a number of investigators have produced transgenic (Tg) mice that express one or all the viral genes. This review attempts to catalogue and characterize the Tg mice produced to date. The topics addressed are HBV, HCV and HDV gene expression and regulation; HBV replication models and factors that inhibit replication; HBV pathogenesis models; HBV tolerance and persistence models; modulation of the immune response to HBV proteins in Tg mice; T cell receptor Tg mice; and models of hepatocellular carcinoma. PMID- 11064316 TI - Small bowel review: part I. AB - In the past year, there have been many advances in the area of small bowel physiology and pathology. More than 1500 papers were assessed in preparation for this review. Some were selected and reviewed, with a particular focus on presenting clinically useful information for the practising gastroenterologist. Relevant review articles have been highlighted, and important clinical learning points have been stressed. The topics are varied in scope, and wherever possible show a logical progression from basic physiology to pathophysiology to clinical disorders and management. PMID- 11064317 TI - [Structural and biochemical characteristics of pathogenic fungus: cell walls, lipids and dimorphism, and action modes of antifungal agents]. AB - Cell walls (0.1-0.5 microm in thickness) of dermatophytes, at least Trichophyton mentagrophytes and Epidermophyton floccosum, are built of microfibrils (20 nm in diameter) and matrix embedding the fibrils. These fibrils are composed of chitin (70-80%) and a small amount of glucans, and the matrix is composed of beta-1-3, beta1-6 glucan, glucomannan, galactomannan and peptides. Another characteristic structure is the outermost layer (20-50 nm in thickness) of the cell wall, which consists of hydrophobic protein rodlets. Lipids are thought to play important roles in the regulation of dimorphism and virulence in pathogenic fungus. Generally, the ratio of phospholipid/ergosterol is less than 1 in yeast form and 2-20 in mycelial form cells in Candida albicans and Sporothrix schenckii. During the transition from yeast to mycelial forms, phosphatidylinositol and phosphatidylserine are reduced, whereas phosphatidylcholine increases. Phospho lipase D is activated on this transition. Phospholipase B is now known to be a virulence factor in C. albicans. Polyene antifungal agents bind to ergosterol in membrane to form complexes, which generate pores and destroy the structures and functions of membrane. Azole antifungal agents inhibit the synthesis of ergosterol leading to deficiency in ergosterol content in membrane, and impair the function of membranes in fungal cells. We show the effects of polyenes on the ultrastructure of fungal plasma membrane and impairment of ionomycin-induced calcium influx in T. mentagrophytes, so that we can compare the differences in mode of actions between these two groups of agents. PMID- 11064318 TI - Protective antigens and mechanisms of anti-Candida immunity. AB - Life threatening fungal diseases are now frequent in a substantial fraction of the immunocompromised host population. The toxicity and the relative scarcity of efficacious antifungal drugs highlight the need for developing alternative or integrative immunoprophylactic and therapeutic tools; among them the need to develop prophylactic or therapeutic vaccines against candidiasis, a widespread mucosal or deep-seated infection caused primarily by the fungus Candida albicans, are of clear priority. Vaccination is a highly beneficial medical practice, and probably the most cost-effective measure against disease onset and progression. It is based on the use of microbial antigens capable of conferring protection in a susceptible target host. To date, only a handful of Candida albicans antigens have been produced and very few of them have been thoroughly investigated for immunogenicity and protection in experimental models of candidiasis. Thus, approaches to the molecular, biochemical and functional characterization of novel C. albicans encoded molecules are most welcome to improve the perspective of developing in the near future an effective vaccine against C. albicans. Identification of anti-Candida vaccine candidates must take into account the diversity of Candida diseases, the various underlying mechanisms of protection as well as the major immune dysfunctions observed as predisposing factors for disease. Antigens to be considered possible vaccine candidates include members of the aspartyl proteinase (Sap2) family and the 65kDa mannoprotein (MP65) antigen. An additional molecule of C. albicans which has not yet been identified but deserves great consideration as a vaccine candidate is the yeast-killer toxin receptor (KTR). Initial experimental evidence strongly suggests that the above antigens are able to elicit protective immunity against mucosal and/or systemic candidiasis. A series of molecular, biochemical and immunological studies aimed at validating and strengthening this initial evidence are in progress, with the ultimate goal of producing recombinant or natural antigens that can be assessed for their ability to elicit a protective immunity in animal models and the mechanisms whereby protection is achieved, with emphasis on determination of immune correlates of protection. PMID- 11064319 TI - [New developments in therapy of deep mycoses]. AB - Over the past two decades the incidence of deep mycoses caused by several major groups of fungal pathogens such as Candida spp., aspergilli, Cryptococcus neoformans and zygomycetes has risen steadily. Moreover, opportunistic fungal infections due to Fusarium spp., Trichosporon spp., Pseudallescheria boydii and other emerging pathogens, as well as fluconazole-resistant Candida albicans, all of which are often resistant to existing antifungal drugs, are also encountered more and more frequently. This makes it more difficult for the clinician to achieve successful treatment. Thus there is an urgent need to develop new antifungal agents or formulations with advantages over and/or complimentary to existing drugs. This review focuses on current approaches to antifungal chemotherapy with special reference to the clinical development of new drugs, including (ii) lipid formulations of amphotericin B, (i) second-generation azoles and (iii) antifungal lipopeptides. PMID- 11064320 TI - [Transformation of the fungus ball type pulmonary aspergillosis]. AB - Though the concept of semi-invasive pulmonary aspergillosis was advocated in 1981 by Gefter et al., its histopathological appearance has not yet been reported in detail. Pathological studies on fungus ball type pulmonary aspergillosis (PA) were originally made mainly in regard to related bronchi. Chronic-progressive destructive changes cannot be completely explained from this viewpoint alone. Clinically, since bloody sputum and hemoptysis appeared frequently, further studies on the pulmonary vasculature were considered necessary. In the resected lungs of 3 cases of semi-invasive pulmonary aspergillosis, the pathological features of pulmonary vasculature were characterized by numerous fungal clots within pulmonary arteries and veins, marked destruction of pulmonary blood vessels and extensive intravascular fibrin deposition. Intravascular fibrin deposition causes stasis of blood flow, promotes intravascular proliferation of aspergilli and probably accelerates pulmonary destruction caused by blood stasis. Important pathological findings of fungus ball type pulmonary aspergillosis of the semi-invasive subtype with clinical aspects of chronic-progressive lung destruction caused by severe inflammation, were reported for both the vascular and the bronchial system. PMID- 11064321 TI - [Recent knowledge allowing diagnosis and treatment of deep-seated trichosporonosis]. AB - Deep-seated trichosporonosis is a lethal opportunistic infection in immunocompromised patients. For the rapid diagnosis of this condition, we developed a novel nested-PCR assay that detects DNA specific for clinically important strains of Trichosporon in serum of patients with disseminated trichosporonosis. In this assay, two sets of oligonucleotide primers were derived from the sequence of 26 S ribosomal RNA genes of T. asahii. The specific fragment was amplified from T. asahii and T. mucoides but not from other microorganisms. In a retrospective study using serum samples of patients with disseminated trichosporonosis, the specific fragment was detected in 64% (7 of 11). To treat this infection, we studied the efficacy of rhG-CSF alone and in combination with antifungal agents against disseminated trichosporonosis in neutropenic mice. The results suggested that rhG-CSF might be a useful immunomodulator against Trichosporon infections and the therapeutic outcome might be better when used in combination with antifungal agents. PMID- 11064322 TI - [Mycological and serological diagnosis of cryptococcosis]. AB - Methods for the diagnosis of cryptococcosis have been established, including serotyping and serodiagnosis. Slide agglutination tests with factor sera, the phenol oxidase test, and the growth test at 37C are used for rapid identification of Cryptococcus neoformans. We identified 140 strains and found that 86, 10, and 4% of the isolates were serotypes A, D, and A-D, respectively. Twelve of 14 serotype D strains were isolated from cutaneous cryptococcosis. The most reliable method of serodiagnosis is the latex agglutination (LA) test for detection of polysaccharide antigen combined with protease pretreatment. The LA test is also used for prognosis. The clearance of cryptococcal polysaccharide antigen often takes a few years after treatment. To model the persistence of cryptococcal polysaccharides, we examined the clearance of antigen from the blood of rabbits injected with polysaccharide. The distribution and elimination half-lives of the antigen suggest the prolonged survival of C. neoformans. Recently, the number of cases of C. albidus and C. laurentii has been increasing. The antigenic pattern and the sensitivity of C. albidus in the LA test are the same as that of C. neoformans serotype A. In contrast, C. laurentii does not react with factor sera for C. neoformans and the reactivity with sensitized latex is extremely low. These results were supported by the chemical structures of polysaccharides from these species. We should consider non-neoformans cases in both the identification of isolates and in serodiagnosis. PMID- 11064323 TI - [Molecular epidemiology of Sporothrix schenckii]. AB - Restriction fragment length polymorphism (RFLP) in mitochondrial DNA (mtDNA) of clinical and environmental isolates of Sporothrix schenckii was investigated. Among mtDNA RFLP patterns with Hae III, of 14 environmental isolates morphologically identified as S. schenckii, only 2 isolates were confirmed as S. schenckii, while of more than 500 clinical isolates, all were confirmed to have this condition. Therefore, RFLP analysis of mtDNA is essential for the identification of environmental, but not clinical, isolates of S. schenckii. Isolates of Sporothrix schenckii were classified into 23 mtDNA types (Types l-23) based on mtDNA RFLP patterns with HaeIII and clustered into two major groups by phylogeny, Group A (Types 1-3, 11, 14-19, 22 and 23) and Group B (Types 4-10, 12, 13, 20 and 21). Group A isolates are predominant in South Africa, North America, Central America and South America, while Group B isolates are predominant in Australia and Japan. In Japan, the relative distribution of the mtDNA types varied with geographic region: Types 4, 6 are comparatively abundant in West Japan (Kansai and Kyushu districts), Type 5 is comparatively abundant in East Japan (Tokai, Kanto and Tohoku districts) and Type 2 is abundant in the Hokuriku district. Type 1 is found only in the Hokuriku district. PMID- 11064324 TI - [Detection of Emericella nidulans from bedding materials in horse breeding environment and its significance as a causative agent of guttural pouch mycosis in horses]. AB - Sixty-six new and used samples of horse bedding materials: 60 rice straws, 2 wheat straws, 2 timothy hays and 2 wood chips, were collected from horse breeding stables of 33 farms in Japan and examined for the presence of Emericella nidulans (E. nidulans; anam. Aspergillus nidulans). The incidence of E. nidulans in the bedding materials was 75.8% and there was no significant difference in detection of the fungus between the new and used materials (25 out of the 33 samples, respectively). The growth of E. nidulans isolated in sterilized rice straw culture was accelerated by the addition of water up to about Aw 0.94, which as determined to be the most favorable moisture content. The addition of 0.3% urea solution onto the sterilized rice straw culture also appeared to very effectively enhance its conidial and ascocarp formation. A significant influence of urea on conidial and ascocarp formation of E. nidulans isolates was confirmed by their cultures on a synthetic medium which had urea as the sole nitrogen. These results suggest that severe contamination of E. nidulans on new bedding materials can be hazardous and its proliferation can readily occur at the stable due to the enhancing effect of urine. This analysis is meaningful to elucidate a reservoir of E. nidulans as the causative agent of guttural pouch mycosis in horses. PMID- 11064325 TI - Characterization of an extracellular keratinase from Microsporum canis. AB - Extracellular keratinase (Ekase) 48-, 34- and 31.5-kDa polypeptides, which were isolated from Microsporum canis and examined by immunoblotting reacted with a monoclonal antibody against Ekase of M. canis. We analyzed the amino acid and determined the first 17 amino acid NH2-terminal sequences of the 48-, 34- and 31.5-kDa polypeptides. These polypeptides had a high aspartic acid, glycine and alanine content, respectively. The first 17 amino acid residues of the 34-kDa polypeptide were homologous to those of thermomycolin. This indicated that the 34 kDa polypeptide of Ekase is homologous to the thermomycolin produced by Malbranchea pulchella. Furthermore, Ekase was very heat-stable in the presence of 50 mM CaCl2 at 55 degrees C, since 50% of the initial activity remained. In contrast, no activity was detected after heating in the absence of CaCl2. These results indicate a close relationship between dermatophytes and M. pulchella. PMID- 11064326 TI - Two cases of tinea corporis by infection from a rabbit with Arthroderma benhamiae. AB - The first cases of tinea corporis with Arthroderma benhamiae in Japan are reported. A 7-year-old girl and a 30-year-old mother in Shimane prefecture suffered from dermatophyte infections on the neck, shoulder, arms and leg. Three isolates from the two patients and a rabbit by which they supposedly were infected, were identified as Trichophyton mentagrophytes. On the bases of mating tests using the tester strains of both the African race and the Americano European race of A. benhamiae, they were identified as A. benhamiae African race mating type (-). Our results are the first to indicate that both races of A. benhamiae exist in Japan. PMID- 11064327 TI - [Two cases of Trichophyton mentagrophytes infection contracted from a hamster and a chinchilla]. AB - We report two cases of Trichophyton mentagrophytes infection. Case 1: A 10-year old girl visited Tokyo Electric Power Hospital in June 1994 for evaluation of an erythematous lesion on her head. Three months of topical steroid therapy exacerbated the lesion with pustular formation. Histopathological and mycological examination revealed that the patient had tinea capitis caused by T. mentagrophytes. T. mentagrophytes was also isolated from her pet, a hamster. Case 2: A-14-year-old girl was referred to Shonan Clinic in January 1996 with scaly erythema on her face. She had been treated with neticonazole hydrochloride at another clinic, but the lesion became worse. Direct microscopic examination of the scale was negative at that time, so treatment with topical steroid was started. After 10 days, the lesion was almost cured, but one month later it recurred with an annular distribution. KOH preparation of the scale revealed mycelia and T. mentagrophytes was isolated on culture. T. mentagrophytes was also isolated from her pet, a chinchilla. In both cases, the oral administration of itraconazole at 50 mg/day was effective. The isolated pathogen was identified as Arthroderma vanbreuseghemii with species-specific primers of chitin synthase 1 gene. T. mentagrophytes is one of the most common dermatophytes isolated from man and animals. Rodents like the hamster and the chinchilla have recently become popular as pets in Japan. We should be aware that rodents may carry this kind of fungal pathogen as they become even more popular as pets. PMID- 11064328 TI - Receptor-mediated targeting of gene delivery vectors: insights from molecular mechanisms for improved vehicle design. AB - One way to deliver transgenes to cells in a selective manner is to target the delivery vehicles, or vectors, to specific cell-surface receptors as a first step toward ultimate transport of the gene to the nucleus for expression. While selective delivery, although often to undesired cell types, occurs naturally for some viral vectors and can be achieved for nonviral vehicles, current understanding and control of the delivery mechanism is inadequate for many therapeutic applications. The complicated nature of receptor-mediated transgene uptake and transport requires improved analysis to more effectively evaluate delivery vehicles. As receptor-mediated pathways for gene delivery typically involve vector binding, internalization, subcellular trafficking, vesicular escape, nuclear translocation, and unpackaging for transcription, each of these processes offer mechanisms that can be exploited to enhance targeted gene delivery via properly designed vehicles. For the purpose of this review, current targeted gene delivery vehicles are divided into three approaches: viral, synthetic, and hybrid vectors. Each approach possesses advantages as well as disadvantages at the present time for in vitro and in vivo application, and provides particular challenges to overcome in order to gain significantly improved targeted delivery properties. Quantitative experiments and mathematical modeling of the gene delivery pathway will serve to provide insight into molecular mechanisms and rate-limiting steps for effective gene expression. Information on molecular mechanisms obtained by such methodologies can then be applied to specific vectors, whether viral, synthetic, or hybrid, allowing for the creation of targeted, effective, and safe gene therapeutics. PMID- 11064329 TI - Effects of pore size in 3-D fibrous matrix on human trophoblast tissue development. AB - The effects of pore size in a 3-D polyethylene terephthalate (PET) nonwoven fibrous matrix on long-term tissue development of human trophoblast ED27 cells were studied. Thermal compression was used to modify the porosity and pore size of the PET matrix. The pore size distributions in PET matrices were quantified using a liquid extrusion method. Cell metabolic activities, estradiol production, and cell proliferation and differentiation were studied for ED27 cells cultured in the thermally compressed PET matrices with known pore structure characteristics. In general, metabolic activities and proliferation rate were higher initially for cultures grown in the low-porosity (LP) PET matrix (porosity of 0.849, average pore size of 30 microm in diameter) than those in the high porosity (HP) matrix (porosity of 0.896, average pore size of 39 microm in diameter). However, 17beta-estradiol production and cell differentiation activity in the HP matrix surpassed those in the LP matrix after 12 days. The expression levels of cyclin B1 and p27kip1 in cells revealed progressively decreasing proliferation and increasing differentiation activities for cells grown in PET matrices. Also, difference in pore size controlled the cell spatial organization in the PET matrices and contributed to the tissue development in varying degrees of proliferation and differentiation. It was also found that cells grown on the 2 D surface behaved differently in cell cycle progression and did not show increased differentiation activities after growth had stopped and proliferation activities had lowered to a minimal level. The results from this study suggest that the 3-D cell organization guided by the tissue scaffold is important to tissue formation in vitro. PMID- 11064330 TI - Biofilm thickness variability investigated with a laser triangulation sensor. AB - Measurement of the surface roughness and thickness of biological films is laborious and usually destructive, thus hampering research in this area. We developed a laser triangulation sensor (LTS) set-up for the fast and nondestructive measurement of these biofilm parameters during growth. Using LTS measurements, the morphological development of a dichloromethane-(DCM) degrading biofilm cultured on a wetted-wall column was studied. The measurements show that the biofilm develops faster at the entrance of the reactor. The biofilm consisted of a base film in which microbial colonies were embedded. The biofilm-surface area gradually increased by 23% compared to the bare surface due to the formation of a large number of these colonies. The number and shape of these colonies were followed in time. Using LTS measurements, biofilms distinctly different in surface roughness could be distinguished in a laboratory trickling filter removing DCM from a waste gas. The consequences of the observed surface characteristics for the reaction-diffusion process in the biofilm and for the falling film hydrodynamics are discussed. PMID- 11064331 TI - Modeling of olive oil degradation and oleic acid inhibition during chemostat and batch cultivation of Bacillus thermoleovorans IHI-91. AB - Olive oil degradation by the thermophilic lipolytic strain Bacillus thermoleovorans IHI-91 in chemostat and batch culture was modeled to obtain a general understanding of the underlying principles and limitations of the process and to quantify its stoichiometry. Chemostat experiments with olive oil as the sole carbon source were successfully described using the Monod chemostat model extended by terms for maintenance requirements and wall growth. Maintenance requirements and biomass yield coefficients were in the range reported for mesophiles. For a chemostat experiment at D = 0.3 h(-1) the model was validated up to an olive oil feed concentration of about 3.0 g L(-1) above which an inhibitory effect occurred. Further analysis showed that the liberated oleic acid is the main cause for this inhibition. Using steady-state oleic acid concentrations measured in chemostat experiments with olive oil as substrate it was possible to derive a kinetic expression for oleic acid utilization, showing that a concentration of 430 mg L(-1) leads to a complete growth inhibition. Oleic acid accumulation observed during batch fermentations can be predicted using a model involving growth-associated lipase production and olive oil hydrolysis. Simulations confirmed that this accumulation is the cause for the sudden growth cessation occurring in batch fermentations with higher olive oil start concentrations. Further, an oscillatory behavior, as observed in some chemostat experiments, can also be predicted using the latter model. This work clearly demonstrates that thermophilic lipid degradation by Bacillus thermoleovorans IHI 91 is limited by long-chain fatty acid beta-oxidation rather than oil hydrolysis. PMID- 11064332 TI - Stability and activity of alcohol dehydrogenases in W/O-microemulsions: enantioselective reduction including cofactor regeneration. AB - Microemulsions provide an interesting alternative to classical methods for the conversion of less water-soluble substrates by alcohol dehydrogenase, but until now stability and activity were too low for economically useful processes. The activity and stability of the enzymes are dependent on the microemulsion composition, mostly the water and the surfactant concentration. Therefore, it is necessary to know the exact phase behavior of a given microemulsion reaction system and the corresponding enzyme behavior therein. Because of their economic and ecologic suitability polyethoxylated fatty alcohols were investigated concerning their phase behavior and their compatibility with enzymes in ternary mixtures. The phase behavior of Marlipal O13-60 (C13EO6 in industrial quality)/cyclohexane/water and its effect on the activity and stability of alcohol dehydrogenase from Yeast (YADH) and horse liver (HLADH) and the carbonyl reductase from Candida parapsilosis (CPCR) is presented in this study. Beside the macroscopic phase behavior of the reaction system, the viscosity of the system indicates structural changes of aggregates in the microemulsion. The changes of the enzyme activities with the composition are discussed on the basis of transitions from reverse micelles to swollen reverse micelles and finally, the transition to the phase separation. The formate dehydrogenase from Candida boidinii was used for the NADH-regeneration during reduction reactions. While the formate dehydrogenase did not show any kinetic effect on the microemulsion composition, the other enzymes show significant changes of activity and stability varying the water or surfactant concentration of the microemulsion. Under certain conditions, stability could be maintained with HLADH for several weeks. Successful experiments with semi-batch processes including cofactor regeneration and product separation were performed. PMID- 11064333 TI - Effect of beta-galactosidase hydration on alcoholysis reaction in organic one phase liquid systems. AB - Alcoholysis reactions were performed in organic one-phase liquid systems with E. coli beta-galactosidase to produce heptyl-beta-galactoside from lactose and 1 heptanol. The reaction rate was highly dependent on the amount of water solubilized in the alcohol. A larger amount of water led to a system of two liquid phases in which the alcoholysis rate was 73% faster than in the one-phase system. No hydrolysis reaction of either lactose or product was observed in one phase liquid systems up to 20 h, independent of the water content. Solubility of lactose in the organic phase increased with the water content in the system and the reaction followed the Michaelis-Menten model. Water activity was calculated for heptanol containing different amounts of water and the obtained values were used to estimate the hydration of beta-galactosidase from known models. Enzyme activity correlated with sorbed water, similar to the behavior reported for lysozyme in low water environments. It is concluded that water contribution to enzyme hydration dominates the rate of reaction compared to its effect on lactose solubilization. PMID- 11064334 TI - Equilibrium position, kinetics, and reactor concepts for the adipyl-7-ADCA hydrolysis process. AB - One of the building blocks of cephalosporin antibiotics is 7-amino deacetoxycephalosporanic acid (7-ADCA). It is currently produced from penicillin G using an elaborate chemical ring-expansion step followed by an enzyme-catalyzed hydrolysis. However, 7-ADCA-like components can also be produced by direct fermentation. This is of scientific and economic interest because the elaborate ring-expansion step is performed within the microorganism. In this article, the hydrolysis of the fermentation product adipyl-7-ADCA is studied. Adipyl-7-ADCA can be hydrolyzed in an equilibrium reaction to adipic acid and 7-ADCA using glutaryl-acylase. The equilibrium reaction yield is described as a function of pH, temperature, and initial adipyl-7-ADCA concentration. Reaction rate equations were derived for adipyl-7-ADCA-hydrolysis using three (pH-independent) reaction rate constants and the apparent equilibrium constant. The reaction rate constants were calculated from experimental data. Based on the equilibrium position and reaction rate equations the hydrolysis reaction was optimized and standard reactor configurations were evaluated. It was found that equilibrium yields are high at high pH, high temperature and low-initial adipyl-7-ADCA concentration. The course of the reaction could be described well as a function of pH (7-9), temperature (20-40 degrees C) and concentration using the reaction rate equations. It was shown that a series of CSTR's is the best alternative for the process. PMID- 11064335 TI - "Oxidative stress" response in submerged cultures of a recombinant Aspergillus niger (B1-D). AB - A recombinant strain of Aspergillus niger (B1-D), engineered to produce the marker protein hen egg white lysozyme, was investigated with regard to its susceptibility to "oxidative stress" in submerged culture in bioreactor systems. The culture response to oxidative stress, produced either by addition of exogenous hydrogen peroxide or by high-dissolved oxygen tensions, was examined in terms of the activities of two key defensive enzymes: catalase (CAT) and superoxide dismutase (SOD). Batch cultures in the bioreactor were generally found to have maximum specific activities of CAT and SOD (Umg x protein(-1)) in the stationary/early-decline phase. Continuous addition of H2O2 (16 mmole L(-1) h( 1)), starting in the early exponential phase, induced CAT but did not increase SOD significantly. Gassing an early exponential-phase culture with O2 enriched (25 vol%) air resulted in increased activities of both SOD and CAT relative to control processes gassed continuously with air, while gassing the culture with 25 vol% O2 enriched air throughout the experiment, although inducing a higher base level of enzyme activities, did not increase the maximum SOD activity obtained relative to control processes gassed continuously with air. The profile of the specific activity of SOD (U mg CDW(-1)) appeared to correlate with dissolved oxygen levels in processes where no H2O2 addition occurred. These findings indicate that it is unsound to use the term "oxidative stress" to encompass a stress response produced by addition of a chemical (H2O2) or by elevated dissolved oxygen levels because the response to each might be quite different. PMID- 11064336 TI - Effective production of a thermostable alpha-glucosidase from Sulfolobus solfataricus in Escherichia coli exploiting a microfiltration bioreactor. AB - A microfiltration (MF) membrane bioreactor was developed for an efficient production of a recombinant thermostable alpha-glucosidase (rSsGA) from Sulfolobus solfataricus MT-4. The aim of the membrane bioreactor was to improve the control of the concentration of key components in the growth of genetic engineered microorganisms, such as Escherichia coli. The influence of medium composition was studied in relation to cell growth and alpha-glucosidase production. The addition of components such as yeast extract and tryptone resulted in a higher enzyme production. High cell density cultivation of E. coli BL21(DE3) on semidefined medium, exploiting a microfiltration bioreactor, was studied in order to optimize rSsGA production. In addition to medium composition, the inducer employed (either isopropyl beta-D-thiogalactopyranoside or lactose), the induction duration, and the cultivation mode influenced both the final biomass and the enzyme yield. The MF bioreactor allowed a cell concentration of 50 g/L dry weight and a corresponding alpha-glucosidase production of 11,500 U/L. The improvement obtained in the enzyme production combining genetic engineering and the microfiltration strategy was estimated to be 2,000-fold the wild-type strain. PMID- 11064337 TI - Two parameters account for the flocculated growth of microbes in biodegradation assays. AB - Microbes in activated sludge tanks mostly occur in flocs rather than in cell suspensions. Flocculation results in a limited supply of substrate to the bacteria inside the flocs, which reduces the biodegradation rate of organic compounds by several orders of magnitude. This article presents a simple two parameter extension of growth models for cell suspensions to account for the ensuing reduction of the degradation rate. The additional parameters represent floc size at division and diffusion length. The biomass of small flocs initially increases exponentially at a rate equal to that of cell suspensions. After this first phase, the growth rate gradually decreases and finally the radius becomes a linear function of time. At this time flocs are large and have a kernel of dead biomass. This kernel arises when the substrate concentration decreases below the threshold level at which cells are just able to pay their maintenance costs. We deduce an explicit approximative expression for the interdivision time of flocs, and thereby for the growth of flocculated microbial biomass at constant substrate concentrations. The model reveals that the effect of stirring on degradation rates occurs through a reduction of the floc size at division. The results can be applied in realistic biodegradation quantifications in activated sludge tanks as long as substrate concentrations change slowly. PMID- 11064338 TI - Analysis of oxygenation reactions in a multi-substrate system-A new approach for estimating substrate-specific true yields. AB - A series of experiments was performed in an aerobic chemostat reactor using a multi-substrate system consisting of acetate, phenol, and 2,4-dichlorophenol (DCP). The phenolic compounds require initial oxygenation reactions, while acetate is oxidized without oxygenations. The biomass completely dechlorinated DCP and utilized all of the substrates simultaneously as electron donors and carbon sources. However, DCP removal was less than for phenol and depended on the solids retention time. A novel substrate-specific yield analysis indicated that true yield values were approximated well by the number of electrons removed in non-oxygenation reactions. Experiments for estimating the kinetic parameters for utilization of the phenolic compounds were designed to eliminate the effects of the key cosubstrates of oxygenation reactions, O2, and the reduced intracellular electron carrier, NADH + H+. The maximum specific rate of substrate utilization, qmax, and the half-maximum rate concentration, K, for phenol and DCP were estimated. The kinetics for DCP were much slower than those for phenol, and the largest effect was a half-maximum rate concentration, which was 19 times larger for DCP. The larger K for DCP explains why DCP removal was low and sensitive to the solids retention time. PMID- 11064339 TI - Aerobic degradation of mixtures of chlorinated aliphatics by cloned toluene-o xylene monooxygenase and toluene o-monooxygenase in resting cells. AB - Recombinant strains of Escherichia coli constitutively expressing toluene-o xylene monooxygenase (ToMO) of Pseudomonas stutzeri OX1 and toluene o monooxygenase (TOM) of Burkholderia cepacia G4 were investigated for their ability to oxidize trichloroethylene (TCE), 1,1-dichloroethylene (1,1-DCE), cis 1,2-dichloroethylene (cis-DCE), trans-1,2-dichloroethylene (trans-DCE), vinyl chloride (VC), and chloroform (CF), individually as well as in various mixtures. ToMO oxidized all of these individual compounds well, whereas TOM did not degrade VC significantly (16-fold less) and degraded cis-DCE and trans-DCE less well (3.7 and 2.4-fold, respectively). For mixtures of these chlorinated aliphatics, ToMO was again more robust than TOM. For example, in binary mixtures including TCE, ToMO degraded all three DCE isomers and CF, but the presence of TCE inhibited VC degradation; TOM degraded both TCE/1,1-DCE and TCE/trans-DCE, but not cis-DCE for TCE/cis-DCE, and the addition of CF or VC to TCE completely inhibited degradation of both compounds and TCE. The addition of CF or trans-DCE stimulated VC degradation in the presence of TCE for ToMO, and the addition of any of the three DCE isomers stimulated VC degradation for TOM. Significant degradation of all ternary mixtures of TCE and less chlorinated ethenes, as well as a mixture of TCE, three DCEs, and VC, was achieved with ToMO (but not TOM). In mixtures of these chlorinated compounds, degradation was found to occur simultaneously rather than sequentially, and the mineralization of many of these compounds could be confirmed through detection of chloride ions. PMID- 11064340 TI - Trehalose delays the reversible but not the irreversible thermal denaturation of cutinase. AB - The effect of trehalose (0.5 M) on the thermal stability of cutinase in the alkaline pH range was studied. The thermal unfolding induced by increasing temperature was analyzed in the absence and in the presence of trehalose according to a two-state model (which assumes that only the folded and unfolded states of cutinase were present). Trehalose delays the reversible unfolding. The midpoint temperature of the unfolding transition (Tm) increases by 4.0 degrees C and 2. 6 degrees C at pH 9.2 and 10.5, respectively, in the presence of trehalose. At pH 9.2 the thermal unfolding occurs at higher temperatures (Tm is 52.6 degrees C compared to 42.0 degrees C at pH 10.5) and a refolding yield of around 80% was obtained upon cooling. This pH value was chosen to study the irreversible inactivation (long-term stability) of cutinase. Temperatures in the transition range from folded to unfolded state were selected and the rate constants of irreversible inactivation determined. Inactivation followed first order kinetics and trehalose reduced the observed rate constants of inactivation, pointing to a stabilizing effect on the irreversible inactivation step of thermal denaturation. However, if the contribution of reversible unfolding on the irreversible inactivation of cutinase was taken into account, i.e., considering the fraction of cutinase molecules in the reversible unfolded conformation, the intrinsic rate constants can be calculated. Based on the intrinsic rate constants it was concluded that trehalose does not delay the irreversible inactivation. This conclusion was further supported by comparing the activation energy of the irreversible inactivation in the absence and in the presence of trehalose. The apparent activation energy in the absence and in the presence of trehalose were 67 and 99 Kcal/mol, respectively. The activation energy calculated from intrinsic rate constants was higher in the absence (30 Kcal/mol) than in the presence of trehalose (16 Kcal/mol), showing that kinetics of the irreversible inactivation step increased in the presence of trehalose. In fact, trehalose stabilized only the reversible step of thermal denaturation of cutinase. PMID- 11064341 TI - Production of luciferase-magnetic particle complex by recombinant Magnetospirillum sp. AMB-1. AB - Luciferase-bacterial magnetic particle (BMP) complexes were produced by recombinant Magnetospirillum sp. AMB-1. We constructed plasmids pKML and pNELM, respectively, by fusing luc to the 5' and 3' terminal of magA, encoding an integral iron translocating protein situated in the BMP membrane, of AMB-1. In addition, we produced bifunctional active-fusion proteins on BMPs by using a plasmid pAcML. In this plasmid, acetate kinase and luciferase genes were fused to the N-terminus and the C-terminus of MagA, respectively. Bacterial magnetic particles isolated from transconjugants for pKML, pNELM and pAcML exhibited luciferase activity. Bacterial magnetic particles isolated from transconjugants for pAcML also exhibited acetate kinase activity. Fed-batch culture of pKML transconjugant yielded 2.6 mg BMPs per liter of culture, and 95% conversion of iron into magnetite was obtained, at a nitrate concentration of 1.4 mM. Continuous feeding of iron as ferric quinate significantly enhanced growth and total magnetic production. Final cell concentration of 1.8 x 10(9) cells/mL and 6 mg per liter of culture was obtained. Magnetite production by fed-batch culture of AMB-1 was about 3 times that obtained by batch culture. There were no significant differences in BMPs yield between recombinant AMB-1 cultivated by fed batch culture and wild type of AMB-1. PMID- 11064342 TI - Localization of initial lymph node metastasis from carcinoma of the thoracic esophagus. AB - BACKGROUND: Most surgeons consider esophageal carcinoma with lymph node involvement a systemic disease. However, it is possible that the disease may be localized in the earlier phases of lymphatic metastasis. The distribution of involved lesions in the initial phase of lymph node metastasis has not been thoroughly investigated yet. METHODS: Among 329 patients that underwent curative (R0 International Union Against Cancer [UICC]) esophagectomy with systematic mesoesophageal dissection, 51 cases of patients with only 1 involved lymph node (solitary involvement) were retrospectively investigated and compared with patients with multiple involved lymph nodes. The regional lymph nodes were divided into the thoracocervical junction group (lower deep cervical and recurrent nerve lymph nodes), perigastric group, and intrathoracic group. RESULTS: Lymph node involvement was limited to a solitary lymph node in 46% of lymph node positive patients with esophageal carcinoma confined to the wall (T1 and T2, UICC) and in 17% of lymph node positive patients with cancer that invaded the extramural layer (T3 and T4, UICC). Of patients with solitary involvement, 82% had a positive thoracocervical junction or perigastric lymph node. The 5-year survival rate in solitary involvement cases was 61%, and 65% when solitary involvement was not intrathoracic. Most of the 5-year survivors had involvement of a thoracocervical junction or perigastric lymph node and had not received systemic chemotherapy. CONCLUSIONS: Solitary involvement was not rare and not directly associated with a disseminated disease. Solitary involvement was commonly located in the thoracocervical junction or abdomen that are accessible without thoracotomy. Systematic dissection of the regional lymph nodes including thoracocervical junction and perigastric groups is recommended for resectable esophageal carcinoma at this time. However, less extensive dissection may be performed in selected cases if the sentinel lymph node concept proves valid. PMID- 11064343 TI - Frequency and clinical impact of lymph node micrometastasis and tumor cell microinvolvement in patients with adenocarcinoma of the esophagogastric junction. AB - BACKGROUND: Tumor involvement of regional lymph nodes has a crucial impact on the prognosis of patients with adenocarcinoma of the esophagogastric junction (AEG). Although additional tumor cell deposits can be detected by sensitive methods (e.g., immunohistochemistry and polymerase chain reaction), their prognostic significance is uncertain. METHODS: Using immunohistochemistry for cytokeratins (AE1/AE3 antibody), the authors studied 3987 regional lymph nodes from 145 patients with completely resected adenocarcinoma of the esophagus (AEG I; n = 46 patients), cardia (AEG II; n = 79 patients), and subcardial region (AEG III; n = 20 patients). The newly detected cells were categorized with tumor cell microinvolvement (TCM) or with micrometastases (MM) based on tumor size and histology. RESULTS: Of the 75 pathologic lymph node negative (pN0) patients, 3 of 30 patients in the AEG I group (10%) and 8 of 45 patients in the AEG II and III groups (18%) had TCM (no significant difference). MM was found in 2 of 30 tumors in the AEG I group (7%) and in 11 of 45 tumors in the AEG II and III groups (24%), a significantly lower rate that that in the AEG I group (P < 0.05). Neither TCM nor MM showed a significant prognostic impact in AEG I tumors (P > 0.05). For the AEG II and III tumors, MM (new lymph node positive [pN+] cases) had a prognostic impact similar to metastases found by routine methods, with reclassification based on MM resulting in improvement in the pN0 group from 72.8 months to 82.6 months, but almost no change was seen in the pN+ group (49.9-49.2 months). TCM had no adverse impact on survival in any tumor type. CONCLUSIONS: These results highlight important differences between AEG I tumors and AEG II and III tumors and argue for different lymphadenectomy strategies for patients with these tumor types. PMID- 11064344 TI - Asian patients with gastric carcinoma in the United States exhibit unique clinical features and superior overall and cancer specific survival rates. AB - BACKGROUND: The 5-year survival rate from gastric carcinoma, stratified by stage, is markedly greater in the Far East than in the United States. This survival rate advantage may reflect differences in diagnostic criteria, more complete staging, more radical surgery, or less aggressive tumor biology. METHODS: A historic cohort of consecutive cases of gastric carcinoma reported to the population-based California Cancer Registries of Orange, San Diego and Imperial Counties from 1984 to 1996 was studied. Factors associated with Asian race were profiled using logistic regression. Multivariate survival analyses were performed using a Cox proportional hazard model. RESULTS: Two thousand four hundred sixteen patients (64%) were non-Latino white; 690 (18%) were Latino; 94 (2.5%) were black; 541 (14%) were of Asian descent: Korean (22%), Vietnamese (20%), Japanese (20%), Chinese (14%), and Filipino (12%). Asian patients were more likely to have localized (lymph node negative) disease (odds ratio [OR], 1.61; 95% confidence interval [CI], 1.23-2.10), less likely to have tumors of the gastroesophageal junction (OR, 0.22; 95% CI, 0.15-0.31,), and less likely to be older than 50 years (OR, 0.58; 95% CI, 0.43-0.77). Asian patients with gastric carcinoma were twice as likely as non-Latino whites to be alive at 5 years (20.9% vs. 10.2%; P < 0.0001). Multivariate analyses indicated that whites had an increased risk of dying from all causes (relative risk [RR], 1.34; 95% CI, 1.16-1.55; P < 0.01) and of dying from cancer in comparison to Asian patients (RR, 1.26; 95% CI, 1.07 1.48; P < 0.05). CONCLUSIONS: Asians who received a diagnosis of gastric carcinoma in the United States have less advanced disease than non-Asians. The increased proportion of localized disease and improved survival rates of patients of Asian descent in the United States is consistent with less aggressive tumor biology. PMID- 11064345 TI - Low dose fractionated radiation enhances the radiosensitization effect of paclitaxel in colorectal tumor cells with mutant p53. AB - BACKGROUND: The current study was undertaken to investigate the influence of wild type or mutant p53 status on the radiosensitizing effect of paclitaxel in colorectal tumor cell lines. METHODS: HCT-116 (contains wild-type p53) and HT-29 (contains mutant p53) established from moderately differentiated colorectal carcinomas were used in this study. Colony-forming assay was performed after exposure to either different radiation doses (0.5-6 gray [Gy]) or paclitaxel (1 10 nM) or in combination. Induction of p53 and p21(waf1/cip1) by these treatments were determined by immunocytochemistry and Western blot analysis. RESULTS: Radiation caused an increase in nuclear p53 and p21(waf1/cip1) proteins in HCT 116 cells, indicating that p53 functionally induced p21(waf1/cip1). However, induction of nuclear p53 and p21(waf1/cip1) protein was not evident in HT-29 cells, suggesting that p53 was not functional in these cells. Survival data showed that the HCT-116 cells (survival fraction of exponentially growing cells that were irradiated at the clinically relevant dose of 2 Gy [SF(2)] = 0.383; dose required to reduce the fraction of cells to 37% [D(0)] = 223 centigray [cGy]) were significantly sensitive to ionizing radiation (P < 0.008) when compared with the HT-29 cells (SF(2) = 0.614; D(0) = 351 cGy). Paclitaxel caused a higher degree of clonogenic inhibition in HCT-116 (D(0) = 0.7 nM) than HT-29 (D(0) = 1.11 nM) cells (P < 0.06). When paclitaxel and radiation were combined, an enhanced radiosensitizing effect (P < 0.05) was observed in HCT-116 cells (SF(2) = 0.138; D(0) = 103 cGy), whereas in HT-29 cells no significant radiosensitization of paclitaxel was observed (SF(2) = 0.608; D(0) = 306 cGy). However, pretreatment with paclitaxel followed by multifractionated low dose radiation (0.5- or 1-Gy fractions for a total dose of 2 Gy) significantly enhanced the radiosensitizing effect in both HCT-116 and HT-29 cells. CONCLUSIONS: The results of the current study suggested that multifractionated radiation given at very low doses after exposure of cells to paclitaxel conferred a potent radiation sensitizing effect irrespective of p53 status. PMID- 11064346 TI - Classification of advanced colorectal carcinomas by tumor edge morphology: evidence for different pathogenesis and significance of polypoid and nonpolypoid tumors. AB - BACKGROUND: Increasing evidence suggests that a substantial proportion of colorectal carcinomas develop without a preexisting polypoid adenomatous lesion, but it is difficult to detect the possible origin of advanced carcinomas. The purpose of this study was to test the validity and significance of a new histopathologic classification system based on the histologic analysis of the tumor edge. METHODS: One hundred eighty-six unselected cases of colorectal carcinoma were included. A new classification method to distinguish polypoid and nonpolypoid growth type was based on the presence or absence of elevation of tumor as compared with adjacent mucosa. Inter- and intraobserver agreement of classification was tested. Association with other clinicopathologic features including histopathologic characteristics of the tumors, presence or absence of lesional and concurrent adenoma, K-ras mutations, and prognosis was evaluated. RESULTS: Classification could be made in 75% of the tumors, and 25% were unclassifiable, mostly due to absence of tumor margin in sections. Of the classifiable carcinomas, 45% were classified as polypoid, of which 52% had lesional adenoma. Nonpolypoid tumors formed 48% of classifiable cases, and only 2% had lesional adenoma. Features of both polypoid and nonpolypoid carcinomas were present in 7% of cases. Concurrent extralesional adenomas were found more frequently in association with polypoid carcinomas. K-ras mutations were more common in polypoid (43%) than in nonpolypoid tumors (8%; P = 0.018). Nonpolypoid carcinomas were significantly (P = 0.03) more aggressive than polypoid carcinoma, with 38% and 20% recurrence rates, respectively. CONCLUSIONS: The authors' results indicate that advanced colorectal carcinomas can be classified according to growth pattern by observing the tumor edge. This classification has prognostic significance because nonpolypoid carcinomas appeared to have a worse prognosis than polypoid ones. PMID- 11064347 TI - Enhanced expression and activation of Ca(2+)/calmodulin-dependent protein kinase IV in hepatocellular carcinoma. AB - BACKGROUND: Ca(2+)/calmodulin-dependent protein kinase IV (CaM-kinase IV) is a multifunctional protein kinase that is expressed abundantly in the central nervous system and, to a lesser degree, in nonneuronal tissues such as the liver. In the current study, the authors demonstrated the expression of CaM-kinase IV in hepatocytes from hepatocellular carcinoma (HCC) in both humans and rats. METHODS: Immunoblotting and immunohistochemical analysis were performed to confirm the expression of CaM-kinase IV and CaM-kinase kinase in HCC occurring in both humans and rats. The kinase activity of CaM-kinase IV in the lysate of each of these liver supernatant fluids was measured using a specific substrate (peptide gamma) for this enzyme before and after phosphorylation by exogenously added CaM-kinase kinase. RESULTS: Marked positive staining of HCC hepatocytes was found and the subcellular staining pattern mainly was cytosolic. One immunoreactive band with a molecular weight of 64 kilodaltons, which was identical to an isoform of rat cerebellum CaM-kinase IV, was demonstrated by immunoblotting. Ca(2+)/calmodulin dependent CaM-kinase IV activity was high in HCC and showed almost no difference in activity in specimens with and without CaM-kinase kinase phosphorylation. CONCLUSIONS: CaM-kinase IV was found to be expressed in HCC and might have been involved in the development of HCC. CaM-kinase IV that was expressed in cancerous hepatocytes was phosphorylated mainly by CaM-kinase kinase that also was expressed in tumor cells. PMID- 11064348 TI - Cost analysis of pancreatic carcinoma treatment. AB - BACKGROUND: Pancreatic carcinoma is a major health issue and financial burden to society. To improve the quality and efficiency of care delivered, it is essential for health care providers to have a good understanding of the cost of treatment. METHODS: The authors examined the facility-based costs and survival of 103 patients with pancreatic carcinoma who were treated at the Karmanos Cancer Institute between January 1992 and September 1998. Longitudinal cost data for each patient were obtained, and from those data, 6-month, 1-year, and lifetime total treatment costs were calculated. RESULTS: The average 6-month, 1-year, and lifetime total treatment costs were $37,327, $42,218, and $48,803, respectively, and the median survival was 7 months. In univariate analyses, the disease stage at diagnosis was a highly significant predictor of total cost. Patients with metastatic disease had the lowest cost, and patients with resectable disease had the highest cost. In multivariate analyses controlling for disease stage, treatment strategies and dual insurance coverage were also important predictors of costs but patient age, race, and gender were not predictive. Disease stage also was highly predictive of survival. In a multivariate analysis controlling for disease stage, chemotherapy and radiation therapy were correlated with longer survival, whereas resection and palliative bypass surgery were not. CONCLUSIONS: The costs of treating patients with pancreatic carcinoma are considerable, even though survival duration typically is short. Disease stage was the most dominating factor determining costs and survival. After controlling for disease stage, chemotherapy, surgery, and dual insurance coverage were also significantly associated with higher cost of care. However, in survival analyses, only chemotherapy and radiation therapy were associated with a significant increase in patient survival. PMID- 11064349 TI - Combined therapy with conservative surgery, radiotherapy, and regional chemotherapy for maxillary sinus carcinoma. AB - BACKGROUND: Current goals for the treatment of carcinoma of the maxillary sinus include the preservation of vision, ability to eat, ability to communicate, and appearance as well as cure. METHODS: Seventy-five Japanese patients who presented with maxillary sinus carcinoma between 1979 and 1997 were analyzed retrospectively. There were 48 males and 27 females with a median age of 62 years. The mean follow-up period was 73 months. All patients underwent multimodality therapy including surgery through a sublabial incision, radiotherapy, and regional chemotherapy. The regional lymph nodes were treated only in those patients with cervical lymph node involvement. RESULTS: The 5-year and 10-year overall survival rates were 76% and 66%, respectively. In 65 patients with squamous cell carcinoma, the 5-year and 10-year overall survival rates were 77% and 66%, respectively. All 23 patients with orbital involvement retained the orbital contents and 17 patients demonstrated adequate ocular function. There was no disease recurrence reported among patients with involvement of the foramen rotundum or the foramen ovale, whereas two of the three patients with invasion of the foramen lacerum developed disease recurrence. There were 12 complications in 12 patients, including double vision (4 patients), cataracts (3 patients), trismus (4 patients), and fistula formation (1 patient). CONCLUSIONS: Control of the primary tumor site is important in the curative treatment of patients with maxillary sinus carcinoma. Combined therapy with conservative surgery, radiotherapy, and regional chemotherapy appears to be an effective method for local control and the preservation of ocular function. PMID- 11064350 TI - Association of allelic imbalance at locus D13S171 (BRCA2) and p53 alterations with tumor kinetics and chromosomal instability (aneuploidy) in nonsmall cell lung carcinoma. AB - BACKGROUND: BRCA2 gene, located at chromosome 13q12.3, frequently is altered in familial types of cancer in which a "double-hit" inactivation model seems to occur. In contrast, in sporadic forms of cancer there is frequent absence of a second event (point mutations) suggesting that allelic imbalance at the BRCA2 locus may be associated with a "gene dosage effect" of BRCA2 function. Little is known about BRCA2 allelic alterations in nonsmall cell lung carcinomas (NSCLCs). Furthermore, recent studies suggest that BRCA2 and p53 participate in a common pathway involved in DNA damage repair. In view of this putative link, the authors investigated in a series of 63 NSCLCs: 1) the allelic imbalance (AIm) at the D13S171 (BRCA2) locus, 2) the possible relation with tumor kinetics (proliferation [PI] and apoptotic indices [AI]) and chromosomal instability (aneuploidy) of the carcinomas, and 3) the mutual impact of D13S171 AIm and p53 altered status on the above-mentioned parameters. METHODS: Allelic status of the BRCA2 region was examined in a series of 63 NSCLCs, by using the polymorphic marker D13S171, which is located in the center of it. Most information regarding the status of p53 at the immunohistochemical and genetic levels was obtained from a previous analysis. Tumor kinetic parameters (proliferation and apoptotic indices) were determined using Ki-67 immunohistochemical analysis and Tdt mediated dUTP nick end labeling assay, respectively. Chromosomal instability (aneuploidy) was assessed by measuring nuclear DNA ploidy with an image analysis system. RESULTS: Allelic imbalance at D13S171(BRCA2) was observed in 70% of the informative cases (H: heterozygous) with a rather high frequency of occurrence (50%) in Stage I disease, suggesting a possible early involvement in the development of NSCLCs. Although no association was found among loss of heterozygosity (LOH) at D13S171, kinetic parameters and ploidy status of the tumors and concurrent alterations in BRCA2 and p53 (BRCA2[LOH]/p53[P]), which was the most frequent profile (37.2%), had the highest growth index (PI/AI mean value ratio) that differed significantly only from the BRCA2(LOH)/p53(N) pattern (P = 0.027). This difference was attributed to the high AI of the BRCA2(LOH)/p53(N) pattern (P < 0.001), whereas PI was similar among all BRCA2/p53 profiles. Also the "full abnormal pattern" was associated with aneuploidy, whereas the BRCA2(LOH)/p53(N) profile was mainly diploid. When these indicators and conventional prognostic ones were examined for effect on patient survival, only stage and lymph node status showed a significant correlation, whereas LOH at D13S171 (BRCA2), p53 abnormalities, proliferative and apoptotic indices, ploidy status, smoking history, and histology and combinations of LOH and p53 abnormalities failed to show significant correlation with survival. CONCLUSIONS: These findings suggest that in BRCA2(LOH) NSCLCs the status of p53 (wild type or mutant) represents a decisive determinant of tumor growth and chromosomal instability. Nevertheless, a possible synergistic effect from loss of D13S171 region with p53 abnormalities cannot be excluded because the BRCA2(LOH)/p53(P) profile compared with the BRCA2(H)/p53(P) one had a higher PI/AI mean value ratio (31.05 vs. 22.97), although it was not statistically significant. However, we cannot exclude the possibility that LOH at D13S171 reflects deletion of other putative tumor suppressor gene(s) in the proximity of BRCA2. In this respect, more studies are needed to understand the involvement of BRCA2 region alterations in nonsmall cell lung carcinogenesis. (c) 2000 American Cancer Society. PMID- 11064351 TI - High dose radiation therapy and chemotherapy as induction treatment for stage III nonsmall cell lung carcinoma. AB - BACKGROUND: The current study was conducted to review the authors' experience in treating consecutive patients with American Joint Committee on Cancer (1997 revision) Stage III nonsmall cell lung carcinoma with aggressive preoperative chemoradiation followed by surgical resection. METHODS: The records of all patients who received preoperative chemoradiation were evaluated. Patients received 2 cycles of concurrent cisplatin and etoposide with 5940 centigrays of radiation therapy. They then were reevaluated to determine whether they were surgical candidates. If so, resection of the primary tumor with mediastinal lymph node dissection was performed 4-6 weeks after the completion of preoperative treatment. After adequate healing, an additional four cycles of cisplatin/etoposide or carboplatin/paclitaxel was given. RESULTS: Forty-two patients received preoperative chemoradiation, 33 of whom underwent surgical resection (79%), including 9 patients who underwent pneumonectomies. Complete pathologic responses were observed in 27% of these patients. Postoperative complications were noted in 21% of the patients and included persistent air leak, supraventricular arrhythmia, and empyema. There were no reported treatment related deaths. The median follow-up was 26 months. The overall 5-year survival rate for all patients was 36.5% and was 45. 3% for patients who underwent resection. A trend toward increased 5-year survival was observed in patients who had a complete pathologic response (57.1%). Univariate analysis revealed the N stage classification to be significant for predicting a complete response. Patterns of failure revealed the brain to be the most common site of first recurrence (50%) and the only site of recurrence in 36% of patients. There was only one case of local failure. CONCLUSIONS: Preoperative chemoradiation using high radiation doses is feasible with acceptable toxicity. The results of the current study suggest an increased complete pathologic response rate and increased overall survival rate compared with reports in the published literature. PMID- 11064352 TI - Similar outcome of elderly patients in intergroup trial 0096: Cisplatin, etoposide, and thoracic radiotherapy administered once or twice daily in limited stage small cell lung carcinoma. AB - BACKGROUND: Elderly patients comprise a significant portion of patients with limited stage small cell lung carcinoma. However, the prognostic importance of age has been controversial, and concern for toxicity often hinders enthusiasm for offering full dose therapy. METHODS: In this retrospective analysis of Intergroup Trial 0096, the authors compared the outcome of patients 70 years or older to those younger than 70 years. Patients received cisplatin 60 mg/m(2), Day 1 and etoposide 120 mg/m(2), Days 1-3 for 4 cycles and either once or twice daily concurrent thoracic radiotherapy to 45 grays. RESULTS: Of 381 patients, 50 (13%) were age 70 years or older. The elderly group did not differ significantly from those younger than 70 years with respect to gender distribution, performance status, or weight loss. Severe hematologic toxicity (Grade 4-5: 61% vs. 84%; P < 0.01) and fatal toxicity (1% vs. 10%; P = 0.01) occurred more often among older patients. There were no differences in the frequency of nonhematologic toxicities. Response rate (88% vs. 80%; P = 0.11), event free survival rate (5 year, 19% vs. 16%; P = 0.18), time to local failure, and duration of response did not differ between groups. Overall survival rates (5 year, 22% vs. 16%; P = 0.05) favored those younger than 70 years. Much of the difference in overall survival rates between age groups occurred within the first 6 months on study. CONCLUSIONS: Elderly patients had similar response and survival rates compared with those younger than 70 years. However, toxicity, particularly hematologic, was greater among the elderly. Selected older patients, such as those with a good performance status, should be considered for optimum treatment approaches. PMID- 11064353 TI - Reproductive functions in female patients treated with adjuvant and neoadjuvant chemotherapy for localized osteosarcoma of the extremity. AB - BACKGROUND: The side effects of chemotherapy on ovarian and reproductive functions in female patients who received adjuvant and/or neoadjuvant treatment for localized osteosarcoma of the extremities at our institution in the last 21 years (1974-1995) were assessed. METHODS: Ninety-two patients with a mean actual age of 26 (14-51) were interviewed. They had been followed 3 to 16 years after treatment (mean: 9.6 yrs). Twenty-four of them had received chemotherapy before puberty and 68 after puberty. RESULTS: Amenorrhea during chemotherapy occurred in 69% of postpuberal patients but only in 2 patients aged 39 and 43, respectively, was permanent. After the end of treatment, the patients' menstrual activity started again, and only a slight number of cases showed increases of menstrual irregularities. Twenty-two patients married after treatment; 20 patients started a pregnancy at a mean age of 27. Of these 20 patients, 3 had voluntary abortions and 17 succeeded. At the time of article submission, three were pregnant, 14 had 19 full term pregnancies, and no birth defects nor congenital anomalies were registered in their 19 full term newborns. CONCLUSIONS: In this group of female patients, chemotherapy seemed to alter neither their reproductive function nor the health of their newborns. PMID- 11064354 TI - Mechanism for bone invasion of oral cancer cells mediated by interleukin-6 in vitro and in vivo. AB - BACKGROUND: Osteoclastic bone resorption is an important step in bone invasion in several malignancies. Although interleukin (IL)-6 accelerates osteoclastic bone resorption, it remains unclear whether IL-6 may be involved in bone invasion of oral cancer. METHODS: The pit formation assay with calf femur-derived bone slices was performed to examine the bone-resorbing activity of osteoclasts and cancer cells. The chemotaxis activity of the culture media was analyzed by the use of Boyden chamber technique. Nude mice, which were inoculated with IL-6-producing oral cancer cells into masseter, were treated with anti-IL-6 neutralizing antibody, and mandibular-bone invasion of the cells was assessed. RESULTS: BHY, a bone-invasive oral cancer cell line, but not HNT, a noninvasive cell line, produced large amounts of IL-6. In a pit formation assay, addition of conditioned medium (CM) derived from BHY but not HNT increased osteoclastic bone resorption, and the effects were inhibited by anti-IL-6 antibody. BHY-secreted IL-6 showed significant chemotaxis activity for osteoclasts. Of note, CM from the cocultivation of osteoclasts and BHY markedly enhanced the cancer cell migration, and the chemotaxis activity was significantly reduced when anti-IL-6 antibody was added into the coculture and then CM were collected, but not when the antibody was added into the CM after they were collected. Furthermore, treatment with anti IL-6 antibody almost completely inhibited mandibular bone invasion of BHY in nude mice. CONCLUSIONS: These results strongly suggest that IL-6 secreted by oral cancer cells plays a significant role in bone invasion. PMID- 11064355 TI - The prognostic significance of p16(INK4a)/p14(ARF) locus deletion and MDM-2 protein expression in adult acute myelogenous leukemia. AB - BACKGROUND: The p16(INK4a) locus encodes two distinct proteins, p16(INK4a) and p14(ARF). Although p16(INK4a) and p15(INK4b) are involved in the phosphorylation of the retinoblastoma (Rb) protein, p14(ARF) interacts with the MDM-2 oncoprotein antagonizing its function as a suppressor of p53. The role of deletions of p16(INK4a)/p14(ARF) and p15(INK4b) and expressions of MDM-2 in myeloid leukemias and its influence on prognosis remain unclear. METHODS: The authors analyzed deletions of p16(INK4)/p14(ARF) and p15(INK4b) in 74 adults with acute myeloid leukemia (AML) by Southern blotting. Western blotting was used to determine Rb protein phosphorylation in patients with deletions of p16(INK4)/p14(ARF) and p15(INK4b). Then, they analyzed the levels of MDM-2 protein expression and correlated it with prognosis in an expanded population of 79 adults with AML by immunoblot analysis and solid-phase radioimmunoassay. RESULTS: Deletions of p16(INK4a)/p14(ARF) and/or p15(INK4b) occurred in 4 of 74 patients (5%) (hemizygous in 3, homozygous in 1 patient). Although the complete remission (CR) rate was similar (79% vs. 50%; P = 0.187), CR duration (10 vs. 46 weeks; P < 0.001), event free survival rate (EFS; 6 vs. 85 weeks; P < 0.004) and overall survival rate (11 vs. 86 weeks; P = 0.001) were significantly shorter in patients with deletions of p16(INK4a)/p14(ARF) and/or p15(INK4b). Thirty-seven (47%) of 79 patients studied for MDM-2 showed increased MDM-2 expression. These patients had a significantly shorter EFS rate (50 vs. 64 weeks; P = 0.023) and a trend for shorter CR duration (24 vs. 53 weeks; P = 0.07). Overall survival rate was not significantly different (50 vs. 84 weeks; P = 0.136). CONCLUSIONS: The authors concluded that 1) deletions of p16(INK4a)/p14(ARF) and/or p15(INK4b) occur with low incidence in patients with AML; 2) patients with deletions of p16(INK4a)/p14(ARF) and/or p15(INK4b) have a significantly shorter CR duration, EFS rate, and overall survival rate than do patients without deletions; (3) overexpression of MDM-2 is common in AML and is associated with shorter CR duration and EFS rate. Mechanisms other than p14(ARF) deletion are responsible for MDM-2 overexpression, and this overexpression may play a role in the biology of the disease. PMID- 11064356 TI - Surgical therapy for distant metastases of malignant melanoma. AB - BACKGROUND: Manifestation of distant metastases in melanoma patients commonly indicates a poor prognosis. The aim of the current study was to examine the role of surgical treatment in these patients. METHODS: Data from 444 patients with distant melanoma metastases were gathered prospectively from January 1978 through December 1997. Characteristics of the primary tumor, time until the first occurrence of distant metastases, frequency and site of distant metastases, surgical therapy, and survival were evaluated by univariate and multivariate analyses. RESULTS: Histology, Breslow thickness, Clark level, and pT and pN categories (UICC 1997) significantly influenced the median interval from initial diagnosis to manifestation of distant metastases. The most common single localization was the lung (n = 83), followed by distant lymph node (n = 79), and skin metastases (n = 51). One hundred seventy-four patients received surgical treatment (39%) and 111 (25%) patients received surgical treatment with curative resection (R0, UICC 1997), most frequently in distant lymph node or skin metastases (57% and 59%, respectively). Median survival time and 2-year survival rate for all patients were 7 months and 15.8%, respectively, 17 months and 36.1% following curative resection, 6 months and 12.7% after incomplete resection (n = 63) (P < 0.0001). Conservatively treated patients survived for a median of only 4 months with a 2-year survival of 8.1%. Multivariate survival analysis showed localization of the primary tumor (head/neck/trunk vs. extremities), the number of involved sites, and surgical therapy to be independent prognostic factors. CONCLUSIONS: Surgical therapy of distant metastases was most beneficial when complete removal of metastatic tissue was achieved. Selection of patients for surgical excision should be determined by individual patient indications. PMID- 11064357 TI - A reverse transcriptase-polymerase chain reaction assay in the diagnosis of soft tissue sarcomas. AB - BACKGROUND: Many types of sarcomas are characterized by specific chromosomal translocations that result in the production of novel chimeric genes. Detection of these fusion genes could be a sensitive molecular diagnostic assay. However, to the authors' knowledge there have been few systemic comparisons between the current histopathologic diagnosis and the presence or absence of particular fusion genes in patients with adult soft tissue sarcomas (STSs). METHODS: Total RNA was extracted from 75 cases of STS and analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) assay for the detection of a variety of fusion transcripts. The results of the molecular assay were compared with standard histopathologic diagnoses. RESULTS: Of the 18 tumors diagnosed as synovial sarcoma, 17 (94%) expressed SYT-SSX chimeric transcripts. All nine myxoid liposarcomas were positive for FUS-CHOP fusion transcripts. Of the four cases of Ewing sarcoma, two had an EWS-FLI1 fusion transcript and one had an EWS-ERG fusion transcript. A clear cell sarcoma had a EWS-ATF1 fusion transcript. None of 19 cases of malignant fibrous histiocytoma nor 3 leiomyosarcomas contained a fusion transcript. Three cases with an initial diagnosis other than synovial sarcoma expressed a SYT-SSX fusion transcript. A review of the slides and additional examination showed that a diagnosis of synovial sarcoma was appropriate for these cases. There was a trend for biphasic synovial sarcoma to contain the SYT-SSX1 fusion. CONCLUSIONS: The authors believe RT-PCR assay for the detection of a specific fusion gene provides a useful tool for confirmation of the diagnosis of adult STS in diagnostically difficult cases and in retrospective studies. PMID- 11064358 TI - Breast carcinoma after cancer at another site: method of detection, tumor characteristics, and surgical treatment. AB - BACKGROUND: As the number of cancer survivors increases, so will the number of second primary cancers, including breast carcinoma after cancer at another site. Limited information is available regarding the clinical characteristics of breast carcinoma after a primary at another site. METHODS: TUMORS (The Upper Midwest Oncology Registry Services) was used to identify 937 women with breast carcinoma occurring as a second primary after a first primary at a known site other than the breast. They were compared with a sample of 1874 women with first primary breast carcinoma, frequency-matched by age to the second primary group, for method of detection, tumor characteristics, and type of surgery. RESULTS: Women with breast carcinoma after cancer at another site tended to have smaller tumors and less extensive disease than women with first primary breast carcinoma and were somewhat more likely than first primary cases to have had their breast carcinoma detected by mammogram or clinical breast exam rather than detecting it themselves. Differences in method of detection accounted for differences in tumor size and extent. Second primary breast carcinoma was less likely to be lobular or mixed ductolobular carcinoma compared with first primary breast carcinoma. Surgical treatment (mastectomy vs. breast-conserving surgery) did not differ for first and second primary breast carcinoma. CONCLUSIONS: Clinical characteristics of breast carcinoma after cancer at another site were by and large similar to those of first primary breast carcinoma. The more favorable prognostic characteristics among women with a history of cancer were accounted for by increased medical surveillance. PMID- 11064359 TI - The prognostic significance of p53 tumor suppressor gene alterations in ovarian carcinoma. AB - BACKGROUND: The prognostic significance and nature of p53 dysfunction in ovarian carcinoma is unclear. The relation between p53 overexpression, p53 mutations, and their effects on overall survival in primary ovarian carcinoma is explored. METHODS: Tumor specimens from 171 consecutive epithelial ovarian carcinomas were examined for overexpression of p53 protein with DO7 antibody. P53 mutations were determined by direct sequencing. The influences of conventional histopathologic prognostic factors and various p53 molecular alterations on overall survival were assessed. RESULTS: Overall, 48.5% and 57.3% of the samples showed p53 overexpression and p53 mutation, respectively. Although neither p53 overexpression nor the mere presence of a p53 mutation impacted overall survival, the combination did prognosticate survival both in univariate and multivariate models. The authors' results suggest 4 mechanisms that may affect p53 dysfunction in nearly 100% of advanced stage ovarian carcinomas. These include null mutation, nonresponsive p53 (wild-type [wt] p53 sequence, DO7 negative), sequestration (wt p53 sequence, DO7 positive), and missense mutation. Median survival for these groups that constitute sequentially 21.3%, 20.5%, 12.3%, and 45.9% of the 122 Stage III or IV (International Federation of Gynecology and Obstetrics) cancers was 1.49, 1.31, 3.09, and 3.6 years, respectively. The nonresponsive p53 and null sequence tumors grouped together as functionally null convey the worst prognosis relative to missense mutations in a univariate model (P = 0.006). Functionally null p53 (P = 0.002), stage (P = 0.008), and optimal cytoreduction (P = 0.008) were independent prognostic factors by multivariate analysis. CONCLUSIONS: Sequestration of wt p53 is unique to advanced stage ovarian carcinoma. Functionally null p53 represents an independent molecular predictor of compromised survival. PMID- 11064360 TI - Increasing levels of hypoxia in prostate carcinoma correlate significantly with increasing clinical stage and patient age: an Eppendorf pO(2) study. AB - BACKGROUND: The purpose of this study was to analyze the extent of hypoxia in prostate carcinoma tumors using the Eppendorf pO(2) microelectrode and correlate this with pretreatment characteristics and prognostic factors. METHODS: Custom made Eppendorf pO(2) microelectrodes were used to obtain pO(2) measurements from the pathologically involved region of the prostate (as determined by the pretreatment sextant biopsies) as well as from a region of normal muscle for comparison. Each set of measurements comprised approximately 100 separate readings of pO(2), for a total of 10,804 individual measurements. Fifty-five patients with localized prostate carcinoma were studied: Forty-one patients received brachytherapy implants, and 14 patients underwent radical prostatectomy. The pO(2) measurements were obtained in the operating room by using a sterile technique under spinal anesthesia for the brachytherapy group and under general anesthesia for the surgery group. The Eppendorf histograms were recorded and described by the median pO(2), mean pO(2), and percentage < 5 mm Hg and < 10 mm Hg. A multivariate mixed-effects analysis for the prediction of tumor oxygenation was performed and included the following covariates: type of tissue (prostate vs. muscle), type of treatment (implant vs. surgery) and/or anesthesia (spinal vs. general), prostate specific antigen level, disease stage, patient age and race, tumor grade, tumor volume, perineural invasion, and hormonal therapy. RESULTS: Due to differences in patient characteristics and the anesthesia employed, control measurements were obtained from normal muscle (in all but two patients). This internal comparison showed that the oxygen measurements from the pathologically involved portion of the prostate were significantly lower (average median pO(2), 9.9 mm Hg) compared with the measurements normal muscle (average median pO(2), 28.6 mm Hg; P < 0.0001). A multivariate, linear, mixed analysis demonstrated that, among all of the patients, the significant predictors of oxygenation were tissue (prostate vs. muscle) and anesthesia (spinal vs. general) or treatment (implant vs. surgery). Among the brachytherapy (spinal anesthesia) patients, the significant predictors of pO(2) were tissue type, disease stage, and patient age. There were no significant predictors of oxygenation in the surgical (general anesthesia) group. CONCLUSIONS: This study, employing in vivo electrode oxygen measurements, demonstrated that hypoxia exists in prostate carcinoma tumors. A dramatic effect of anesthesia was observed, likely due to modulation of polarography in the presence of fluorine. Within the group of brachytherapy (spinal anesthesia) patients, increasing levels of hypoxia (within prostatic tissue) correlated significantly with increasing clinical stage and patient age. More patients will be accrued to this prospective study to further correlate the oxygenation status in prostate carcinoma tumors with known prognostic factors and, ultimately, treatment outcome. PMID- 11064361 TI - Distribution of mRNAs encoding CRF receptors in brain and pituitary of rat and mouse. AB - Two G protein-coupled receptors have been identified that bind corticotropin releasing factor (CRF) and urocortin (UCN) with high affinity. Hybridization histochemical methods were used to shed light on controversies concerning their localization in rat brain, and to provide normative distributional data in mouse, the standard model for genetic manipulation in mammals. The distribution of CRF R1 mRNA in mouse was found to be fundamentally similar to that in rat, with expression predominating in the cerebral cortex, sensory relay nuclei, and in the cerebellum and its major afferents. Pronounced species differences in distribution were few, although more subtle variations in the relative strength of R1 expression were seen in several forebrain regions. CRF-R2 mRNA displayed comparable expression in rat and mouse brain, distinct from, and more restricted than that of CRF-R1. Major neuronal sites of CRF-R2 expression included aspects of the olfactory bulb, lateral septal nucleus, bed nucleus of the stria terminalis, ventromedial hypothalamic nucleus, medial and posterior cortical nuclei of the amygdala, ventral hippocampus, mesencephalic raphe nuclei, and novel localizations in the nucleus of the solitary tract and area postrema. Several sites of expression in the limbic forebrain were found to overlap partially with ones of androgen receptor expression. In pituitary, rat and mouse displayed CRF-R1 mRNA signal continuously over the intermediate lobe and over a subset of cells in the anterior lobe, whereas CRF-R2 transcripts were expressed mainly in the posterior lobe. The distinctive expression pattern of CRF-R2 mRNA identifies additional putative central sites of action for CRF and/or UCN. Constitutive expression of CRF-R2 mRNA in the nucleus of the solitary tract, and stress-inducible expression of CRF-R1 transcripts in the paraventricular nucleus may provide a basis for understanding documented effects of CRF-related peptides at a loci shown previously to lack a capacity for CRF-R expression or CRF binding. Other such "mismatches" remain to be reconciled. PMID- 11064362 TI - Does long-term physical exercise counteract age-related Purkinje cell loss? A stereological study of rat cerebellum. AB - Physical exercise affects properties of the central nervous system that may increase the brain's ability to counteract degenerative changes. We have previously reported that rats trained from 5 to 23 months of age have less age related decrease in spontaneous motor activity than sham-treated sedentary rats. Each rat ran at a speed of 20 m/min on a horizontal treadmill, for 20 minutes, two times per day, 5 days a week. In the present study we have carried out stereological analyses of the cerebella of the same rats. The total number of Purkinje cells was estimated with the optical fractionator technique, the local volumes of individual Purkinje cells with the planar rotator technique, and the volumes of the cerebellar layers with Cavalieris principle. We found that sedentary aged rats have 11% fewer Purkinje cells and 9% smaller Purkinje cell soma volumes (both 2P = 0.02) than exercised aged rats, and that exercised aged rats have the same number of Purkinje cells as young rats. These findings indicate that the degree of age-associated degenerative changes in parts of the central nervous system is dependent on earlier life style and health habits and may be prevented or delayed by physical exercise. PMID- 11064363 TI - Differential localization of septins in the mouse brain. AB - We have carried out a comparative immunohistochemical study on four members of the septin family, CDCrel-1, Septin6, CDC10, and H5, which are abundantly expressed in the adult mouse brain. We found that each septin showed overlapping but distinct distribution at the levels of light and electron microscopy. CDCrel 1 was abundant in inhibitory presynaptic terminals and associated with GABAergic vesicles in the thalamus, globus pallidus, and cerebellar nuclei. Septin6 was associated with synaptic vesicles in various brain regions, including glomeruli of the olfactory bulb. CDC10 was diffusely expressed in the brain and was localized beneath presynaptic membrane and astroglial processes. H5 was localized in the astroglial processes in some specific brain regions. The differential expression and subcellular localization of these septins indicates that a given neuron or glial cell expresses a specific set of septin monomers and that the resulting septin complexes with distinct compositions may play distinct roles in the brain. PMID- 11064364 TI - Status epilepticus-induced hilar basal dendrites on rodent granule cells contribute to recurrent excitatory circuitry. AB - Mossy fiber sprouting into the inner molecular layer of the dentate gyrus is an important neuroplastic change found in animal models of temporal lobe epilepsy and in humans with this type of epilepsy. Recently, we reported in the perforant path stimulation model another neuroplastic change for dentate granule cells following seizures: hilar basal dendrites (HBDs). The present study determined whether status epilepticus-induced HBDs on dentate granule cells occur in the pilocarpine model of temporal lobe epilepsy and whether these dendrites are targeted by mossy fibers. Retrograde transport of biocytin following its ejection into stratum lucidum of CA3 was used to label granule cells for both light and electron microscopy. Granule cells with a heterogeneous morphology, including recurrent basal dendrites, and locations outside the granule cell layer were observed in control preparations. Preparations from both pilocarpine and kainate models of temporal lobe epilepsy also showed granule cells with HBDs. These dendrites branched and extended into the hilus of the dentate gyrus and were shown to be present on 5% of the granule cells in pilocarpine-treated rats with status epilepticus, whereas control rats had virtually none. Electron microscopy was used to determine whether HBDs were postsynaptic to axon terminals in the hilus, a site where mossy fiber collaterals are prevalent. Labeled granule cell axon terminals were found to form asymmetric synapses with labeled HBDs. Also, unlabeled, large mossy fiber boutons were presynaptic to HBDs of granule cells. These results indicate that HBDs are present in the pilocarpine model of temporal lobe epilepsy, confirm the presence of HBDs in the kainate model, and show that HBDs are postsynaptic to mossy fibers. These new mossy fiber synapses with HBDs may contribute to additional recurrent excitatory circuitry for granule cells. PMID- 11064365 TI - Strain and sex differences in the morphology of the medial preoptic nucleus of mice. AB - The medial preoptic nucleus (MPO), which is involved in sexual and maternal behaviors, contains neuronal clusters that have been described as being sexually dimorphic in size and neuropeptide content in a variety of species. A subnucleus in DBA/2J (D2) inbred mice, called the pars compacta of the MPO (MPOpc), is absent in C57BL/6J (B6) inbred mice (Robinson et al. [1985] J. Neurogenet. 2:381 388). We report here on experiments that further characterize strain and sex differences in medial preoptic morphology in D2 and B6 inbred mice. A prominent MPOpc, located within the caudal part of the MPO and dorsal to the suprachiasmatic nucleus, was present in both male and female D2 animals but was absent from B6 animals. MPOpc neurons were darkly stained for Nissl substance and larger than neurons in the surrounding MPO. In D2 brains, galanin-immunoreactive (-ir), oxytocin-ir, vasopressin-ir, and NADPH diaphorase-positive neurons were concentrated within the MPOpc. Fewer similar neurons in the comparable region of the MPO of B6 mice suggests that the absence of the MPOpc is due to absence of these neurons rather than a less compact organization. In D2 animals, the density of galanin-ir neurons in the MPOpc was sexually dimorphic, with higher numbers of galanin-ir neurons in females. Strain differences in galanin-ir, oxytocin-ir, vasopressin-ir, and NADPH diaphorase staining appeared to be limited to the MPOpc. Cholecystokinin-immunoreactive neurons, which have been reported to be numerous in the sexually dimorphic central subdivision of the MPO of rats, were sparse in the MPO of D2 and B6 mice. Confirmation of the MPOpc as an accessory magnocellular neurosecretory nucleus was obtained by finding labeling of MPOpc neurons after injection of DiI into the posterior pituitary. PMID- 11064366 TI - Up-regulation of CD81 (target of the antiproliferative antibody; TAPA) by reactive microglia and astrocytes after spinal cord injury in the rat. AB - We examined the expression of CD81 (also known as TAPA, or target of the antiproliferative antibody) after traumatic spinal cord injury in the rat. CD81, a member of the tetraspanin family of proteins, is thought to be involved in reactive gliosis. This is based on the antiproliferative and antiadhesive effects of antibodies against CD81 on cultured astrocytes, as well as its up-regulation after penetrating brain injury. CD81 expression following dorsal hemisection of the spinal cord was determined immunohistochemically at time points ranging from 1 day to 2 months postlesion (p.l.). In the unlesioned cord a low background level of CD81 was observed, with the exception of the ependyma of the central canal and the pia mater, which were strongly CD81-positive. One day p.l., CD81 was diffusely up-regulated in the spinal cord parenchyma surrounding the lesion site. From 3 days onward, intensely CD81-positive round cells entered the lesion site, completely filling it by 7 days p.l. Staining with the microglial markers OX-42 and Iba1 revealed that these cells were reactive microglia/macrophages. At this time, no significant CD81 expression by GFAP-positive reactive astrocytes was noted. From the second week onward, CD81 was gradually down-regulated; i.e., its spatial distribution became more restricted. The CD81-positive microglia/macrophages disappeared from the lesion site, leaving behind large cavities. After 2 months, astrocytes that formed the wall of these cavities were strongly CD81-positive. In addition, CD81 was present on reactive astrocytes in the dorsal funiculus distal from the lesion in degenerated white matter tracts. In conclusion, the spatiotemporal expression pattern of CD81 by reactive microglia and astrocytes indicates that CD81 is involved in the glial response to spinal cord injury. PMID- 11064367 TI - Organization of callosal linkages in visual area V2 of macaque monkey. AB - In visual area V2 of the macaque monkey callosal cells accumulate in finger-like bands that extend 7-8 mm from the V1/V2 border, or approximately half the width of area V2. The present study investigated whether or not callosal connections in area V2 link loci that are located at the same distance from the V1/V2 border in both hemispheres. We analyzed the patterns of retrograde labeling in V2 resulting from restricted injections of fluorescent tracers placed at different distances from the V1/V2 border in contralateral area V2. The results show that varying the distance of V2 tracer injections from the V1/V2 border led to a corresponding variation in the location of labeled callosal cells in contralateral V2. Injections into V2 placed on or close to the V1 border produced labeled cells that accumulated on or close to the V1 border in contralateral V2, whereas injections into V2 placed away from the V1 border produced labeled cells that accumulated mainly away from the V1 border. These results provide evidence that callosal fibers in V2 preferentially link loci that are located at similar distances from the V1/V2 border in both hemispheres. Relating this connectivity pattern to the topographic map of V2 suggests that callosal fibers link topographically mirror-symmetrical regions of V2, i.e., callosal fibers near the V1/V2 border interconnect areas representing visual fields on, or close to, the vertical meridian, whereas callosal connections from regions away from the V1/V2 border interconnect visuotopically mismatched visual fields that extend onto opposite hemifields. PMID- 11064368 TI - Regulation of laminin-associated integrin subunit mRNAs in rat spinal motoneurons during postnatal development and after axonal injury. AB - Two important prerequisites for successful axon regeneration are that appropriate extracellular molecules are available for outgrowing axons and that receptors for such molecules are found in the regenerating neuron. Laminins and their receptors in the integrin family are examples of such molecules, and laminin-associated integrin subunits alpha 3, alpha 6, alpha 7, and beta 1 mRNAs have all been detected in adult rat motoneurons. We have here, by use of in situ hybridization histochemistry, examined the normal postnatal development of the expression in motoneurons of these mRNAs and integrin beta 4 mRNA, all of which have been associated with laminin-2. We studied the regulation of these mRNAs, 1-42 days after two types of axotomy in the adult rat (sciatic nerve transection, SNT; ventral root avulsion, VRA) and 1-10 days after SNT in the neonatal animal. During postnatal development, there was a distinct shift in the integrin composition from a stronger expression of the alpha 6 subunit to a very clear dominance of alpha 7 in the adult. All types of axotomy in the adult rat induced initial (1-7 days) large up-regulations of alpha 6, alpha 7 and beta1 subunit mRNAs (250-500%). Only minor changes for alpha 3 mRNA were seen, and beta 4 mRNA could not be detected at all in motoneurons. After adult SNT, the alpha 7 and beta 1 subunits were up-regulated throughout the studied period, and the alpha 6 subunit mRNA was eventually normalized. After VRA, however, the alpha 7 and beta1 levels peaked earlier than after SNT and were normalized at 42 days, whereas alpha 6 mRNA was up-regulated longer than after SNT. Neonatal SNT had much smaller effects on the expression of the studied subunits. The results suggest that an important part of the response to axotomy of motoneurons is to up regulate receptors for laminin. The developmental shift in integrin subunit composition and the various responses seen in the lesion models indicate that different isoforms of laminin play a role in the regenerative response. PMID- 11064369 TI - Cholinergic innervation in adult rat cerebral cortex: a quantitative immunocytochemical description. AB - A method for determining the length of acetylcholine (ACh) axons and number of ACh axon varicosities (terminals) in brain sections immunostained for choline acetyltransferase (ChAT) was used to estimate the areal and laminar densities of this innervation in the frontal (motor), parietal (somatosensory), and occipital (visual) cortex of adult rat. The number of ACh varicosities per length of axon (4 per 10 microm) appeared constant in the different layers and areas. The mean density of ACh axons was the highest in the frontal cortex (13.0 m/mm(3) vs. 9.9 and 11.0 m/mm(3) in the somatosensory and visual cortex, respectively), as was the mean density of ACh varicosities (5.4 x 10(6)/mm(3) vs. 3.8 and 4.6 x 10(6)/mm(3)). In all three areas, layer I displayed the highest laminar densities of ACh axons and varicosities (e.g., 13.5 m/mm(3) and 5.4 x 10(6)/mm(3) in frontal cortex). The lowest were those of layer IV in the parietal cortex (7.3 m/mm(3) and 2.9 x 10(6)/mm(3)). The lengths of ACh axons under a 1 mm(2) surface of cortex were 26.7, 19.7, and 15.3 m in the frontal, parietal, and occipital areas, respectively, for corresponding numbers of 11.1, 7.7, and 6.4 x 10(6) ACh varicosities. In the parietal cortex, this meant a total of 1.2 x 10(6) synaptic ACh varicosities under a 1 mm(2) surface, 48% of which in layer V alone, according to previous electron microscopic estimates of synaptic incidence. In keeping with the notion that the synaptic component of ACh transmission in cerebral cortex is preponderant in layer V, these quantitative data suggest a role for this innervation in the processing of cortical output as well as input. Extrapolation of particular features of this system in terms of total axon length and number of varicosities in whole cortex, length of axons and number of varicosities per cortically projecting neuron, and concentration of ACh per axon varicosity, should also help in arriving at a better definition of its roles and functional properties in cerebral cortex. PMID- 11064370 TI - Expression of thyrotropin-releasing hormone receptor 2 (TRH-R2) in the central nervous system of rats. AB - The distribution of the recently discovered thyrotropin-releasing hormone (TRH) receptor subtype TRH-R2 was studied in rat brain, pituitary, and spinal cord by in situ hybridization histochemistry and compared with the distribution patterns of the other elements of TRH signaling, namely TRH, TRH-R1, and the TRH-degrading ectoenzyme (TRH-DE). In contrast to the very restricted mRNA expression of TRH-R1 in the central nervous system, TRH-R2 mRNA was widely distributed with highest transcript levels throughout the thalamus, in the cerebral and cerebellar cortex, medial habenulae, medial geniculate nucleus, pontine nuclei, and throughout the reticular formation. In accordance with the well-known endocrine function of TRH, TRH-R1 is found predominantly expressed in hypothalamic regions. Expression of TRH-R1 in various brainstem nuclei and spinal cord motoneurons seems to be associated with the described effects of TRH on the vegetative and autonomic system as well as on the somatomotor system. Furthermore, the fully complementary expression of both receptor subtypes, even in regions where transcripts for both receptors were found (e.g., medial septum, lateral hypothalamus superior colliculi, substantia nigra, etc.), indicates that in discrete neuroanatomical pathways the two receptors serve highly specific functions for the transmission of TRH signals. Together with TRH-DE, the putative terminator of TRH actions that shows in various, but not all, brain areas, an overlapping mRNA distribution pattern with both receptors, the distribution of TRH-R2 mRNA seems to provide the anatomical basis for the described effects of TRH on higher cognitive functions as well as its effect on arousal, locomotor activity, and pain perception. PMID- 11064371 TI - Organization and connections of V1 in Monodelphis domestica. AB - We examined the internal organization and connections of the primary visual area, V1, in the South American marsupial Monodelphis domestica. Multiunit electrophysiological recording techniques were used to record from neurons at multiple sites. Receptive field location, size, progressions, and reversals were systematically examined to determine the visuotopic organization of V1 and its boundaries with adjacent visual areas. As in other mammals, a virtually complete representation of the visual hemifield was observed in V1, which was coextensive with a region of dense myelination. The vertical meridian was represented at the rostrolateral boundary of the field, the upper visual quadrant was represented caudolaterally, whereas the lower visual quadrant was represented rostromedially. Injections of fluorescent tracers into V1 revealed dense connections with cortex immediately adjacent to the rostrolateral boundary, in peristriate cortex (PS or V2). Connections were also consistently observed with a caudotemporal area (CT), entorhinal cortex (EC), and multimodal cortex (auditory/visual, A/V). These results demonstrate that M. domestica possess a highly differentiated neocortex with clear functional and architectonic cortical field boundaries, as well as discrete patterns of cortical connections. Some connections of V1 are similar to those observed in eutherian mammals, such as connections with V2 and extrastriate areas (e.g., CT), which suggests that there are general features of visual system organization that all mammals possess due to retention from a common ancestor. On the other hand, connections of V1 with EC and multimodal cortex may be a primitive feature of visual cortex that was lost in some lineages, may be a derived feature of marsupial neocortex, or may be a feature particular to mammals with small brains. PMID- 11064372 TI - Afferent sources to the ganglion of the terminal nerve in teleosts. AB - Afferent sources to the ganglion (ggl) of the terminal nerve (TN) were studied in percomorph teleosts the tilapia and dwarf gourami. After tracer applications to the TN-ggl and the surrounding bulbus olfactorius, retrogradely labeled neurons were present in the area dorsalis telencephali pars posterior (Dp), area ventralis telencephali pars ventralis et supracommissuralis (Vv and Vs), nucleus tegmento-olfactorius of Prasada Rao and Finger (1984), and locus coeruleus. In the contralateral bulbus olfactorius labeled cells were observed, and terminals were seen in the TN-ggl. Tracer injection experiments to the possible sources of origin to the TN-ggl were then performed. Tracer applications to the nucleus tegmento-olfactorius labeled abundant terminals in the TN-ggl but labeled very few in the bulbus olfactorius proper. Retrogradely labeled neurons were present in the nucleus ventromedialis thalami, nucleus commissurae posterioris, area pretectalis pars dorsalis et ventralis, nucleus sensorius nervi trigemini, and formatio reticularis pars superius et medius. Tracer applications to the Dp or Vs/Vv labeled terminals mainly in the bulbus olfactorius proper. However, terminals to the TN-ggl were supplied from labeled axons on their way to the bulbus olfactorius. Tracer injections to the locus coeruleus labeled only a few fibers around the TN-ggl. These results suggest that the TN-ggl receives somatosensory and visual inputs from the nucleus tegmento-olfactorius and olfactory inputs from the bulbus olfactorius and telencephalic subdivisions, which receive secondary olfactory projections. The locus coeruleus may also send fibers to the TN-ggl. PMID- 11064373 TI - Human immune response to recombinant human proteins. AB - Delivery of pharmacological doses of proteins to people has raised concerns of inducing immune responses, especially when the protein is provided in multiple doses over an extended period of time. Immune responses could impact the therapeutic exposure and efficacy of the protein itself. In addition, there have been fears of anaphylaxis or autoimmunity. This review summarizes the available literature regarding the measurement and evaluation of immune responses observed during clinical assessment of recombinant human proteins. Immune responses have ranged from none at all to inactivation and/or accelerated clearance. Presence of antibodies does not necessarily impact therapeutic viability. While responses are related to frequency and route of delivery, there is no clear relationship that enables one to predict the clinical experience. PMID- 11064374 TI - Novel site-specific chemical delivery system as a potential mydriatic agent: formation of phenylephrine in the iris-ciliary body from phenylephrone chemical delivery systems. AB - The objective of this study was to test the three novel ester derivatives of phenylephrone (isovaleryl, phenylacetyl, and pivalyl esters) as potential site specific chemical delivery systems. The mydriatic effect and ocular distribution/metabolism of these compounds were studied by topical application to the eyes of normal rabbits. It was assumed that a reduction-hydrolysis sequence could produce the active phenylephrine in the iris-ciliary body tissues. All the derivatives showed a more pronounced mydriatic effect than that of phenylephrine, whereas phenylephrone was completely devoid of any mydriatic activity. Phenylacetyl ester was the most potent drug, with short duration of action, and showed maximum activity in the presence of 0.01% benzalkonium chloride without causing any visible irritation to the rabbit eye. Administration of the novel compounds to the eyes of the rabbits showed no traces of phenylephrine in the systemic circulation, contrary to topical administration of phenylephrine. Phenylephrone was detected in different compartments of the eye, whereas phenylephrine was present only in the iris-ciliary body tissues following administration of phenylacetyl ester. The conversion of phenylephrone esters to the active drug, phenylephrine, and thus their subsequent activity was dependent on the physicochemical characteristics of the drugs. The results suggest the potential use of phenylacetyl ester as a potent short-term mydriatic agent without systemic side effects. PMID- 11064375 TI - Assessing plasma pharmacokinetics of cholesterol following oral coadministration with a novel vegetable stanol mixture to fasting rats. AB - The purpose of this project was to assess the plasma pharmacokinetics of [(3)H]cholesterol following coadministration of a novel vegetable stanol mixture composed of sitostanol and campestanol (FCP-3P4) to fasting rats. Following an overnight fast (12-16 h) and 48 h post-surgery, adult male Sprague Dawley rats were divided into six treatment groups and received a single-dose oral gavage at 0700 h of either: [(3)H]cholesterol (25 microCi/mL), FCP-3P4 (5 mg/kg) + [(3)H]cholesterol (25 microCi/mL), FCP-3P4 (12.5 mg/kg) + [(3)H]cholesterol (25 microCi/mL), FCP-3P4 (25 mg/kg) + [(3)H]cholesterol (25 microCi/mL), FCP-3P4 (50 mg/kg) + [(3)H]cholesterol (25 microCi/mL), or FCP-3P4 (100 mg/kg) + [(3)H]cholesterol (25 microCi/mL). Intralipid (10%) was the vehicle used to solubilize and coadminister [(3)H]cholesterol and FCP-3P4. Liquid chromatography mass spectrometry analysis confirmed minimal cholesterol and vegetable stanol content within 10% Intralipid. Analysis of plasma pharmacokinetics was initiated by sampling 0.5 mL of blood prior to and 0.25, 0.5 1.0, 2.0, 4.0, 6.0, 8.0, 10, 24, 28, 32, and 48 h post-oral gavage. Plasma samples were obtained by centrifugation of the blood samples and analyzed for [(3)H]cholesterol radioactivity. Pharmacokinetics analysis was performed by standard noncompartmental methods using statistical moment theory. Thin-layer chromatography was used to confirm that the majority of radioactivity measured in plasma was cholesterol (in the form of esterified or unesterified cholesterol). Greater than 90% of the radioactivity measured in all plasma samples was cholesterol-associated (in the form of either esterified or unesterified cholesterol). The coadministration of FCP-3P4 significantly decreased the area under the curve of [(3)H]cholesterol concentration versus time from 0 to 48 h (AUC(0-48h)) and maximum concentration (C(max)) in a dose-dependent manner. However, coadministration of FCP-3P4 at 25, 50, and 100 mg/kg resulted in a significant increase in apparent total body clearance (CL/F, where F is the bioavailability constant), apparent volume of distribution (V(d)/F), and oral absorption rate constant (k(a)) of [(3)H]cholesterol compared with controls. These findings suggest that the novel vegetable stanol mixture, FCP-3P4, modifies the plasma pharmacokinetics of [(3)H]cholesterol in fasting rats on oral coadministration. PMID- 11064376 TI - Studies of the structure of insulin fibrils by Fourier transform infrared (FTIR) spectroscopy and electron microscopy. AB - Fibril formation (aggregation) of insulin was investigated in acid media by visual inspection, transmission electron microscopy (TEM), and Fourier transform infrared (FTIR) spectroscopy. Insulin fibrillated faster in hydrochloric acid than in acetic acid at elevated temperatures, whereas the fibrillation tendencies were reversed at ambient temperatures. Electron micrographs showed that bovine insulin fibrils consisted of long fibers with a diameter of 5 to 10 nm and lengths of several microns. The fibrils appeared either as helical filaments (in hydrochloric acid) or arranged laterally in bundles (in acetic acid, NaCl). Freeze-thawing cycles broke the fibrils into shorter segments. FTIR spectroscopy showed that the native secondary structure of insulin was identical in hydrochloric acid and acetic acid, whereas the secondary structure of fibrils formed in hydrochloric acid was different from that formed in acetic acid. Fibrils of bovine insulin prepared by heating or agitating an acid solution of insulin showed an increased content of beta-sheet (mostly intermolecular) and a decrease in the intensity of the alpha-helix band. In hydrochloric acid, the frequencies of the beta-sheet bands depended on whether the fibrillation was induced by heating or agitation. This difference was not seen in acetic acid. Freeze-thawing cycles of the fibrils in hydrochloric acid caused an increase in the intensity of the band at 1635 cm(-1) concomitant with reduction of the band at 1622 cm(-1). The results showed that the structure of insulin fibrils is highly dependent on the composition of the acid media and on the treatment. PMID- 11064377 TI - Mono-N-carboxymethyl chitosan (MCC), a polyampholytic chitosan derivative, enhances the intestinal absorption of low molecular weight heparin across intestinal epithelia in vitro and in vivo. AB - The synthesis and evaluation of mono-N-carboxymethyl chitosan (MCC) as an intestinal permeation enhancer for macromolecular therapeutics is presented. MCCs were synthesized from two different viscosity grade chitosans to yield both high and low viscosity grade products. These MCCs were tested on Caco-2 cells for their efficiency to decrease the transepithelial electrical resistance (TEER) and to increase the paracellular permeability of the anionic macromolecular anticoagulant low molecular weight heparin (LMWH). For in vivo studies, LMWH was administered intraduodenally with or without MCC to rats. Both types of experiments were performed at pH 7.4. Results show that both viscosity grade MCCs managed to significantly decrease the TEER of Caco-2 cell monolayers when they were applied apically at concentrations of 3-5% (w/v). Transport studies with Caco-2 cells revealed substantial increases of LMWH permeation in the presence of both viscosity grade MCCs compared with controls. In rats, 3% (w/v) low viscosity MCC significantly increased the intestinal absorption of LMWH, reaching the therapeutic anticoagulant blood levels of LMWH. Both in vitro and in vivo results indicate that the polyampholytic chitosan modification MCC is a suitable and functional polymer for the delivery and intestinal absorption of anionic macromolecular therapeutics like LMWH. PMID- 11064378 TI - Regional-dependent intestinal absorption and meal composition effects on systemic availability of LY303366, a lipopeptide antifungal agent, in dogs. AB - Low oral bioavailability and a negative meal effect on drug plasma levels motivated studies on formulation and meal composition effects on the absorption of LY303366, a poorly water-soluble, semisynthetic, cyclic peptide antifungal drug. Solid drug particle size and meal composition studies were evaluated in beagle dogs. Canine regional absorption studies were also carried out utilizing surgically implanted intestinal access ports, and Caco-2 studies were performed to evaluate drug candidate intestinal permeability. Particle size and Caco-2 data indicate that drug permeability limitations to absorption are more important than dissolution rate limits. Caco-2 cell-associated LY303366 approached 10% of incubation concentration that is in the range of the oral bioavailability of the drug. Canine regional absorption studies showed that the extent of LY303366 absorption following duodenal administration was similar to that following oral administration. Significantly lower drug plasma levels were obtained following administration through a colonic access port, a result consistent with poor membrane permeation. Administration of drug with meals of mixed composition, as well as simple fat and protein meals, resulted in significant reductions in AUC(0 48h) compared with results from fasted dogs. In contrast, carbohydrate meals did not reduce drug plasma levels compared to controls. Intravenous pretreatment with devazepide, a cholecystokinin (CCK) antagonist that blocks canine biliary secretion, did not reverse the negative effect of the fat meal on LY303366. Taken together, the results from the present study suggest that membrane-permeability limited absorption is the cause of the observed regionally dependent absorption of LY303366 in the dog and that the observed negative meal effects depend on composition but are independent of biliary secretion. PMID- 11064379 TI - Metal-catalyzed oxidation of human growth hormone: modulation by solvent-induced changes of protein conformation. AB - Metal-catalyzed oxidation (MCO) represents a prominent pathway of protein degradation. To evaluate the importance of the integrity of the metal-binding site on MCO, we subjected recombinant human growth hormone (rhGH), to MCO (ascorbate, Cu(2+), (3)O(2)) in the presence of various aliphatic alcohols (ethanol, ethylene glycol, trifluoroethanol, 1-propanol, 2-propanol, 1,2 propylene glycol, 1-butanol, 2-butanol, and tert-butanol). All alcohols inhibited MCO in a concentration-dependent and sigmoidal manner. Half-points, P(1/2), were dependent on the nature of the alcohol. Circular dichroism and fluorescence spectroscopy were used to monitor cosolvent-induced secondary and tertiary structural changes. The presence of alcohols increased the helical content of rhGH and induced a red shift in the tryptophan emission. The midpoints of the tertiary structural change correlated with the P(1/2) values. Solvent polarity at P(1/2) was determined according to the E(T)(30) scale. All alcohol/water mixtures at P(1/2) had rather similar solvent polarities between 54.5 to 56.4 kcal/mol, with the exception of ethylene glycol. On the other hand, no correlation was obtained between the protection against MCO and the hydroxyl radical-scavenging properties of the cosolvent. We conclude that the primary mechanism of MCO inhibition is a cosolvent-induced conformational perturbation of the metal binding site as opposed to pure radical scavenging. PMID- 11064380 TI - Distribution of gacyclidine enantiomers after experimental spinal cord injury in rats: possible involvement of an active transport system. AB - The pharmacokinetics of gacyclidine enantiomers, a noncompetitive N-methyl-D aspartate (NMDA) antagonist, were studied in plasma and spinal cord extracellular fluid (ECF) after experimental spinal cord injury in rats. Spinal cord trauma was produced by introducing an inflatable balloon in the dorsal subdural space. Upon implantation of microdialysis probes in spinal cord (T9) and intravenous (iv) bolus administration of (+/-)-gacyclidine (2.5 mg/kg), concentrations in plasma and ECF were monitored over 5 h and analyzed by a stereospecific gas chromatography-mass spectrometry (GC-MS) assay. In plasma, concentrations of (+) gacyclidine were approximately 25% higher than those of (-)-gacyclidine over the duration of the experiment and decayed in parallel (t(1/2 alpha) approximately 7 min; t(1/2 beta) approximately 90 min) with no significant difference between the two enantiomers. Clearance (CL) and volume of distribution (Vd) of (-) gacyclidine were approximately 20% higher than those of its optical antipode (CL: 285 versus 236 mL. kg(-1). min(-1); Vd(beta): 39.3 versus 31.2 l/kg). Protein binding (approximately 91%) was not stereoselective. In spinal cord ECF, both enantiomers were quantifiable within 10 min after drug administration, and their concentration remained stable over the duration of the experiment in spite of changing blood concentrations. Repeated iv bolus injections of gacyclidine did not modify these profiles. Areas under the curves (AUCs) of concentration in ECF versus time were similar for both enantiomers and not correlated with AUCs in plasma. Penetration of (-)-gacyclidine was, however, significantly higher (approximately 30%) than that of (+)-gacyclidine. In summary, the disposition of gacyclidine enantiomers is stereoselective. Both enantiomers exhibit a high affinity for spinal cord tissue, and the drug exchange between plasma and spinal cord ECF involves an active transport system. These findings contribute to the explanation of the discrepancy between drug concentrations in plasma and spinal cord ECF. PMID- 11064381 TI - Crosslinking studies in gelatin capsules treated with formaldehyde and in capsules exposed to elevated temperature and humidity. AB - Incomplete in vitro capsule shell dissolution and subsequent drug release problems have recently received attention. A modified USP dissolution method was used to follow capsule shell dissolution, and a 2,4,6-trinitrobenzenesulfonic acid (TNBS) assay was used to follow loss of epsilon-amino groups to study this shell dissolution problem postulated to be due to gelatin crosslinking. The dissolution problems were simulated using hard gelatin capsule (HGC) shells previously treated with formaldehyde to crosslink the gelatin. These methods were also used to study the effect of uncrosslinked HGC stored under stressed conditions (37 degrees C and 81% RH) with or without the presence of soft gelatin capsule shells (SGC). A 120 ppm formaldehyde treatment reduced gelatin shell dissolution to 8% within 45 min in water at 37 degrees C. A 200 ppm treatment reduced gelatin epsilon-amino groups to 83% of the original uncrosslinked value. The results also support earlier reports of non-amino group crosslinking by formaldehyde in gelatin. Under stressed conditions, HGC stored alone showed little change over 21 weeks. However, by 12 to 14 weeks, the HGC exposed to SGC showed a 23% decrease in shell dissolution and an 8% decrease in the number of epsilon-amino groups. These effects on the stressed HGC are ascribed to a volatile agent from SGC shells, most likely formaldehyde, that crosslinked nearby HGC shells. This report also includes a summary of the literature on agents that reduce gelatin and capsule shell dissolution and the possible mechanisms of this not-so-simple problem. PMID- 11064382 TI - Enhanced absorption of insulin and (Asu(1,7))eel-calcitonin using novel azopolymer-coated pellets for colon-specific drug delivery. AB - The objective of this study was to estimate colon-specific delivery of insulin and (Asu(1,7))eel-calcitonin using novel azopolymer-coated pellets. In vitro drug release experiments from the azopolymer-coated pellets containing fluorescein isothiocyanate dextran (MW 4400; FD-4) were carried out by the Japanese Pharmacopoeia (J.P.) XIII rotating basket method with some slight modifications. Little release of FD-4 from the pellets was observed in phosphate buffered saline. However, the release of FD-4 was markedly increased in the presence of rat cecal contents. The intestinal absorption of insulin and (Asu(1,7))eel calcitonin after oral administration of the azopolymer-coated pellets containing these peptides with camostat mesilate was evaluated by measuring the hypoglycemic and hypocalcemic effects, respectively. A slight decrease in plasma glucose levels was observed following the oral administration of these pellets containing 12.5 IU of insulin compared with the same dose of insulin solution. Camostat mesilate, a protease inhibitor that is incorporated with insulin in these pellets, further decreased the plasma glucose levels in a dose-dependent manner. Similar results were also obtained with the oral administration of pellets containing (Asu(1,7))eel-calcitonin. These findings suggest that azopolymer coated pellets may be useful carriers for the colon-specific delivery of peptides including insulin and (Asu(1,7))eel-calcitonin. PMID- 11064383 TI - Note: dissolution of aerosol particles of budesonide in Survanta, a model lung surfactant. AB - The effect of a pulmonary surfactant extract from bovine lung, Survanta, on the dissolution rate of aerosol particles of budesonide was determined. Aerosol particles of budesonide were generated from an ethanol solution, dried, and collected by a cascade impactor for characterization or by a liquid impinger for dissolution experiments. Powder x-ray diffraction, differential scanning calorimetry, differential thermal analysis, and scanning electron microscopy were used to characterize the aerosol particles and starting material. No change in phase was detected, although the aerosol particles appeared to contain residual solvent. The dissolution rate of the aerosol particles in saline was low and variable. Survanta increased the extent of dissolution of budesonide in proportion to the added concentration, which was also verified by equilibrium solubilization studies. Survanta also increased rate of dissolution, in a manner similar to sodium dodecyl sulfate. Analysis of the concentration of budesonide following ultracentrifugation indicated that there is rapid equilibration of budesonide between the Survanta and aqueous phase. These results show that lung surfactant has the potential of enhancing the rate and extent of dissolution of drugs administered to the lung. PMID- 11064384 TI - The essence of mentoring in academic surgery. PMID- 11064385 TI - Preliminary analysis of a randomized clinical trial of adjuvant postoperative RT vs. postoperative RT plus 5-FU and levamisole in patients with TNM stage II-III resectable rectal cancer. AB - OBJECTIVES: Two-hundred eighteen patients with TNM stage II-III resectable rectal cancer, enrolled into a randomized clinical trial, were assessed for efficacy and toxicity of adjuvant postoperative radiation therapy (RT) vs. those of combined RT and chemotherapy (CT), with 5-fluorouracil (5-FU) plus levamisole. End points were overall survival, disease-free survival, the rate of loco-regional recurrence, and treatment-related toxicity. METHODS: Patients in arm I underwent RT (50 Gy) in daily fractions of 2 Gy, 5 days/week for 5 weeks. Patients in arm II began with 5-FU (450 mg/m(2)/day intravenous bolus, days 1-5) plus levamisole (150 mg/day orally, days 1-3); postoperative RT was delivered during week 2 at the same dosage and schedule as in arm I. The other five cycles of CT (5-FU every 28 days and levamisole every 15 days for the length of 5-FU administration) continued after the end of RT if clinical and hemato-biochemical parameters were normal. RESULTS: RT was completed or modified in 170 (90%) of 189 evaluable patients undergoing RT (both treatment groups). Only 44 (59%) of 75 evaluable patients of arm II completed or had an adjustment of the CT schedule; the remaining 31 patients (41%) had to stop or never started the CT regimen. Patients undergoing combined RT and CT had more severe toxicity (enteritis, P = 0.03). There was one CT-related death (gastrointestinal bleeding) in this subset. No significant difference was observed in outcome of patients in the two study groups, nor for pattern of recurrence (heterogeneity chi(2) = 4.82; d.f. = 2; P = 0.08). CONCLUSIONS: These preliminary findings suggest a similar efficacy, coupled with less morbidity, of postoperative RT alone compared with a combined regimen of postoperative RT and CT in patients undergoing radical surgery for stage II-III rectal cancer. PMID- 11064386 TI - Length and quality of survival following external beam radiotherapy combined with expandable metallic stent for unresectable hilar cholangiocarcinoma. AB - BACKGROUND AND OBJECTIVES: Hilar cholangiocarcinoma is a morbid disease with a poor prognosis because resection cannot be performed in many cases. The purpose of this study was to evaluate whether external beam radiotherapy (RT) combined with expandable metallic biliary stent (EMS) affects the length and quality of survival of patients with unresectable hilar cholangiocarcinomas. METHODS: Fifty one patients with unresectable hilar cholangiocarcinoma were retrospectively reviewed. Thirty patients received external beam radiotherapy combined with EMS (EMS+RT group), 10 patients were treated with EMS alone (EMS group), and the remaining 11 patients underwent percutaneous transhepatic biliary drainage alone (PTBD group). The length and quality of survival were analyzed and compared among the three groups. RESULTS: The mean survival of 6.4 months in the EMS group was significantly longer than that of 4.4 months in the PTBD group (P < 0.05). The EMS+RT group with a mean survival of 10.6 months had a significantly longer survival than the EMS group (P < 0.05). The average of the monthly Karnofsky scores of 74.9 in the EMS+RT group and 68.1 in the EMS group, as a parameter of quality of survival, was significantly higher than that of 57.7 in the PTBD group (P < 0.01). The number of hospital days per month of survival was significantly smaller in the EMS+RT and EMS groups than in the PTBD group (10.4, 14.2 vs. 27.3 days; P < 0.001). The EMS+RT group had a longer stent patency than the EMS group (mean: 9.8 vs. 3.7 months; P < 0.001). CONCLUSIONS: These results indicate that external radiotherapy combined with metallic biliary endoprosthesis can increase the length and quality of survival and consequently provide a definite palliative benefit for patients with unresectable hilar cholangiocarcinoma. PMID- 11064387 TI - Palliation of unresectable hilar cholangiocarcinoma. PMID- 11064388 TI - Molecular detection of disseminated cancer cells in the peripheral blood and expression of sialylated antigens in colon cancers. AB - BACKGROUND AND OBJECTIVES: To improve the survival rate of patients with colon cancer, liver metastases must be eradicated in a clinically occult state. This study was designed to find a predictor for potential liver metastases or micrometastases in colon cancer. METHODS: Peripheral blood samples and tumor specimens were obtained from 36 patients with colon cancers. The blood samples were subjected to reverse transcriptase-polymerase chain reaction (RT-PCR) analysis, and the expression of sialylated carbohydrates was also investigated in the tumors immunohistochemically. RESULTS: A carcinoembryonic antigen (CEA) specific signal in the blood was detected in 9 of 12 (75%) patients with liver metastasis and in 8 of 24 (33%) patients without liver metastasis, respectively (P < 0.05). The positive rates of sialyl Lewis A (sLeA) and sialyl Lewis X (sLeX) were 36.3% and 40% in tumors without liver metastasis vs. 58.3% and 100% with liver metastasis, respectively. Within a year after surgery, liver metastases became clinically evident in three of the four patients without liver metastasis who showed a CEA-positive signal in their blood preoperatively and who had tumors with a strong expression of sLeX. CONCLUSIONS: A combination of both markers may provide prognostic information for liver metastases in colon cancer. PMID- 11064389 TI - Clinicopathological characteristics of mucinous carcinoma of the colon and rectum. AB - BACKGROUND AND OBJECTIVES: Clinicopathological significance of colorectal mucinous carcinoma (MC) remains controversial. The aim of the current study was to investigate the clinicopathological characteristics of colorectal MC. METHODS: Eighteen patients with MC and 265 with moderately or well differentiated adenocarcinoma of the colon and rectum, were clinicopathologically compared. RESULTS: MCs occurred in the right colon significantly more frequently than did non-mucinous carcinomas (NMCs). The maximal size of the tumors in MCs (7.0 +/- 2.9 cm) was significantly larger than that in NMCs (5.1 +/- 2.1 cm) (P < 0.001). Although the ratio of patients with peritoneal metastasis in MCs (22.2%; 4/18) was significantly higher than that in NMCs (6.0%; 16/265) (P < 0.05), there was no significant difference regarding liver metastasis. The proportion of lymph node metastasis in MCs (72.2%; 13/18) was significantly higher than that in NMCs (44.9%; 119/265) (P < 0.05). There was no significant difference regarding the lymphatic and venous invasion. The 1-, 3-, and 5-year survival rates of patients with MCs were 77. 8%, 45.4%, and 30.3%, respectively, and were significantly lower than those in patients with NMCs, that were 88.9 %, 65.6%, and 60.8%, respectively (P < 0.05). CONCLUSIONS: As colorectal MCs proliferate and metastasize more rapidly than do NMCs, surgeons should realize that more aggressive surgical treatment should be occasionally administered to improve the postoperative prognosis of the patients with colorectal MCs. PMID- 11064390 TI - Changes of expression level of the differentiation markers in papillary thyroid carcinoma under thyrotropin suppression therapy in vivo immunohistochemical detection of thyroglobulin, thyroid peroxidase, and thyrotropin receptor. AB - BACKGROUND AND OBJECTIVES: Differences in the expression levels of Thyroglobulin (Tg), Thyroid peroxidase (TPO) and thyrotropin receptor (TSH-R) in primary and recurrent specimens under a suppressive serum TSH condition were elucidated in 26 papillary carcinoma patients. METHODS: Immunohistochemical detection was performed by use of each monoclonal antibody against Tg, TPO, and TSH-R. The staining concentrations of the three markers in each specimen were measured for comparison. RESULTS: The mean staining concentrations of Tg, TPO, and TSH-R in the entire primary tumor were 103.92, 104.6 and 89.25, respectively. Five cases showed stronger expression of all the differentiation markers and eight cases showed weaker expression of all these markers in recurrent tissue than in primary tumors. The weaker expression of TSH-R at the recurrent site as compared with that at the primary site significantly demonstrated the shortness of the disease free interval or overall survival. There were significant differences between the death due to cancer and the weaker expression of TSH-R in the recurrent tumor as compared with that in the primary tumor. CONCLUSIONS: Under the TSH suppressive condition, the markers were not expressed uniformly among recurrent tumors. Even under that state, however, low expression of TSH-R in the recurrent tissue was strongly related to a poorer outcome in the patients. PMID- 11064391 TI - Preoperative chemotherapy for esophageal carcinoma with intramural metastasis. AB - BACKGROUND AND OBJECTIVES: The prognosis for patients with intramural metastasis (IMM) of esophageal cancer is poor. We examined the role of preoperative chemotherapy in the management of patients with this disease. METHODS: Fifteen patients with IMM of esophageal carcinoma received preoperative chemotherapy cisplatin on day 1 and 5-fluorouracil on days 1 to 5. This regimen was repeated after a 3-week interval, except in patients with progressive disease or severe toxicity who received only one cycle of chemotherapy. Patients underwent surgery around 3 weeks after completion of chemotherapy. Clinical response was evaluated and survival was compared with that of patients who did not receive preoperative chemotherapy. RESULTS: Toxicity was manageable except in one patient who experienced severe neurological adverse effect. The clinical response rate of the IMM was 66.7% (10/15) and the complete response rate was 6.7% (1/15); for the primary lesion, response rates were 86. 7% and 6.7%, respectively. All 15 patients underwent surgery. Seven of the 15 patients (46.7%) experienced non fatal operative complications. The 5-year survival rate after surgery was 20%. CONCLUSIONS: Preoperative chemotherapy with cisplatin and 5-fluorouracil is feasible in patients with IMM of esophageal carcinoma. This regimen, however, does not improve survival and more effective treatment strategies are required. PMID- 11064392 TI - Direct growth suppressive activity of interferon-alpha and -gamma on human gastric cancer cells. AB - BACKGROUND AND OBJECTIVES: Interferons (IFNs) exhibit anti-tumor activities through either immune modulation or direct anti-tumor effects. We have investigated the activity and mechanisms of IFN-alpha and IFN-gamma on the growth of TSGH9201, TMK-1 and AGS gastric cancer cells in vitro. METHODS: Activities of IFNs on cell growth were analyzed by measuring total cellular DNA. Effects of IFNs on apoptosis was evaluated by formation of in situ DNA breakage and DNA ladders. Effects of IFNs on cells cycle phase distribution were analyzed using flow cytometry. Levels of Bcl-2 family proteins after treatment with IFNs were analyzed using Western blot. RESULTS: Both IFN-alpha and IFN-gamma were active in suppressing the growth of TSGH9201 and TMK-1 cells, while AGS cells were resistant to treatment with IFNs. The IC(50)s of IFN-alpha for TSGH9201 and TMK-1 cells were 300 and 500 U/ml, respectively, and the IC(50)s of IFN-gamma were 40 and 2.0 U/ml, respectively. Both IFN-alpha- and IFN-gamma-induced cell cycle arrest in sensitive cells. IFN-gamma also increased cellular apoptosis, demonstrated by increasing in situ DNA damage and DNA fragmentation. IFN-gamma increased BAK protein levels and decreased Bcl-2 and Bcl-X(S) protein levels in TSGH9201 cells. CONCLUSIONS: IFN-alpha suppressed growth of gastric cancer cells through induction of cell cycle arrest. IFN-gamma suppressed cell growth through induction of both cell cycle arrest and apoptosis. IFN-gamma-mediated apoptosis was associated with the alteration in protein levels of Bcl-2, Bcl-X(S) and BAK. PMID- 11064393 TI - Serum concentrations of soluble interleukin-2 receptor in patients with malignant brain tumors. AB - BACKGROUND AND OBJECTIVES: Interleukin-2 receptor alpha (IL-2Ralpha) can combine with IL-2 firmly, and soluble IL-2Ralpha (sIL-2Ralpha) is elevated in sera from patients with various types of cancers. To investigate the role of this receptor, we studied the changes of serum sIL-2Ralpha in patients with malignant brain tumors. METHODS: SIL-2Ralpha was measured in 100 patients with malignant brain tumors (63 cancer metastasis, 16 malignant gliomas, 21 malignant lymphomas), and 51 patients with cancer who had no distant metastasis such as brain metastasis. RESULTS: In patients with 35 metastatic brain tumors from lung cancer, the levels of sIL-2Ralpha were not significantly different from levels in normal volunteers (311 +/- 62.4 U/ml). In patients with 25 metastatic brain tumors from lung adenocarcinoma, the mean level of serum sIL-2Ralpha was 352 +/- 94.0 U/ml. These same patients showed high levels of serum sIL-2Ralpha (492 +/- 101 U/ml) with regional lymph nodes metastasis. Serum sIL-2Ralpha concentration in 16 patients with malignant glioma varied greatly with the mean concentration of 328 +/- 192 U/ml. In 5 of 16 patients with malignant glioma, we could detect the significant increase of serum sIL-2Ralpha concentration from early stage of recurrence. CONCLUSIONS: Serum levels of sIL-2Ralpha could be a useful immunological marker in patients with malignant brain tumors. PMID- 11064394 TI - Bone metastases from squamous cell carcinoma of the head and neck. AB - BACKGROUND AND OBJECTIVES: Carcinoma of the head and neck is an uncommon primary source of bone metastases. The increasing duration of survival of these patients, however, increases the probability of late bone involvement. The objective was to identify the frequency, clinical presentation, and clinical course of metastatic disease to bone from head and neck primaries. METHODS: A retrospective review was accomplished of the radiographs and nuclear medicine studies for 363 cases of squamous cell carcinoma of the head and neck for whom radiologic studies had been performed. For those with identified bone involvement, a chart review was performed to identify clinical presentation, disease course, and outcome. RESULTS: Only approximately 1% of these patients had clinically demonstrable bone metastases. Eight sites of bone involvement were identified in five patients, including three pelvic, two femoral, and one each humeral, rib, and thoracic spine lesions. All lesions were purely lytic with moth-eaten or permeative borders. Time from primary tumor diagnosis to identification of metastatic disease ranged from being present at diagnosis to a maximum 3.5 years later. Time from identification of metastatic disease to patient death was no greater than 8 months. CONCLUSIONS: Despite the increasing overall survival of patients with these carcinomas, distant bone metastases are infrequent, but should be considered a possibility in any patient with a concurrent or past diagnosis of head and neck carcinoma. The very short time from discovery of bone dissemination to death in most of these patients should be taken into consideration when contemplating operative intervention. PMID- 11064395 TI - Commentary PMID- 11064396 TI - Twelve-year survival after the diagnosis of locally advanced carcinoma of the pancreas: A case report. AB - The long-term survival rate of patients with carcinoma of the pancreas is low. Even more so, long-term survival of patients with metastatic pancreatic carcinoma is extremely rare. In this case report, we describe a patient with an unusual course of disease. This patient was diagnosed with locoregional carcinoma of the pancreas and therefore underwent gastroenterostomy and cholecystojeojenostomy without resection of the primary tumor. Later he was treated with radiotherapy and chemotherapy and survived 12 years, during 11 of which he had no evidence of disease. He died 12 years after the initial diagnosis from peritoneal dissemination of poorly differentiated carcinoma complicated with obstructive jaundice and sepsis. To our knowledge, this patient had the longest reported survival with locally advanced pancreas carcinoma that was not resected. The case is presented and discussed in this article. PMID- 11064397 TI - Intraperitoneal macrophages and tumor immunity: A review. AB - The macrophage (Mphi) is considered the first line of defense in immune response to foreign invaders. Increasing evidence suggests that Mphi(s) also play an important role against neoplastic cells. Mphi(s) exposed to supraphysiologic concentrations of CO(2) are suppressed. As surgeons apply newer minimally invasive techniques to oncologic therapies, it is important to evaluate the impact of these techniques on host-tumor interactions. We review the current understanding of Mphi biology with specific attention on cytotoxicity in addition to tumor immunity. Although systemic immune function is better preserved after laparoscopy than laparotomy, peritoneal Mphi(s) show reduced function after CO(2) pneumoperitoneum than exposure to air. Mphi(s) have shown cytotoxicity to syngeneic cancer cells and may play an important role in tumor surveillance. The impairment in Mphi function after CO(2) exposure may have an effect on outcome after oncologic surgery. In our understanding, Mphi(s) help destroy neoplastic cells. As CO(2) impairs Mphi activity, laparoscopy may significantly alter the host-tumor interaction. PMID- 11064398 TI - Gd-DTPA relaxivity depends on macromolecular content. AB - Gd-DTPA T(1) relaxivity of water protons was measured at 1.5 T and room temperature as a function of macromolecular content in model systems. Gd-DTPA relaxivity was found to increase with macromolecular concentration. The results of this study indicate that the Gd-DTPA relaxivity in tissue extracellular compartment could be as much as 30-70% higher than that of Gd-DTPA in saline. Quantitative MR analyses that use T(1) as an estimation of local Gd-DTPA concentration require a priori determination of the Gd relaxivity in tissue. PMID- 11064399 TI - A new method for fast proton spectroscopic imaging: spectroscopic GRASE. AB - A new fast spectroscopic imaging method is presented which allows both a very short minimum total measurement time and effective homonuclear decoupling. After each excitation, all data points from N(GE) k(x)-k(y)-slices at different k(omega)-values are acquired by using a gradient and spin echo (GRASE) imaging sequence. The delay between consecutive gradient echoes, which are measured with uniform phase encoding between consecutive refocusing alpha-pulses, is the inverse of the spectral width (SW). A refocusing 180 degrees pulse, which is applied within a constant delay between excitation and the GRASE sequence, is shifted in a series of measurements by an increment N(GE)/(2 * SW) to cover the whole k(omega)-k(x)-k(y)-space. Spectroscopic GRASE was implemented on a 4.7 T imaging system and tested on phantoms and normal rat brain in vivo. Measurements were performed with a nominal voxel size of 1.5 x 1.5 x 3 mm(3) and a spatial 64 x 64 matrix. The total measurement time was 2 or 4 min using a repetition time of 1.9 sec, 96 chemical shift encoding steps, SW = 800 Hz, N(GE) = 3, and 2 or 4 accumulations. PMID- 11064400 TI - Spectroscopic imaging of glutamate C4 turnover in human brain. AB - One-dimensional spectroscopic imaging of (13)C-4-glutamate turnover is performed in the human brain with a 6 cc nominal voxel resolution at 4T. Data were acquired with an indirect detection approach using a short spin echo single quantum (1)H (13)C heteronuclear editing method and a 7 cm surface coil with quadrature (13)C decoupling coils. To analyze the data as a function of tissue type, T(1)-based image segmentation through the surface coil was performed to determine the gray and white matter contributions to each voxel. The tricarboxylic acid (TCA) cycle rate in gray and white matter was then determined using a two-compartment model with the tissue fractionation derived from the image segmentation. The mean values for the TCA cycle rate for occipital gray and white matter from three volunteers was 0.88 +/- 0.12 and 0.28 +/- 0.13 respectively, in agreement with literature data. PMID- 11064401 TI - Transit time, trailing time, and cerebral blood flow during brain activation: measurement using multislice, pulsed spin-labeling perfusion imaging. AB - Transit time and trailing time in pulsed spin-labeling perfusion imaging are likely to be modulated by local blood flow changes, such as those accompanying brain activation. The majority of transit/trailing time is due to the passage of the tagged blood bolus through the arteriole/capillary regions, because of lower blood flow velocity in these regions. Changes of transit/trailing time during activation could affect the quantification of CBF in functional neuroimaging studies, and are therefore important to characterize. In this work, the measurement of transit and trailing times and CBF during sensorimotor activation using multislice perfusion imaging with pulsed arterial spin-labeling is described. While CBF elevated dramatically ( thick similar80.7%) during the sensorimotor activation, sizable reductions of transit time ( thick similar0.11 sec) and trailing time ( thick similar0.26 sec) were observed. Transit and trailing times were dependent on the distances from the leading and trailing edges of the tagged blood bolus to the location of the imaging slices. The effects of transit/trailing time changes on CBF quantification during brain activation were analyzed by simulation studies. Significant errors can be caused in the estimation of CBF if such changes of transit/trailing time are not taken into account. PMID- 11064402 TI - Effect of surface coating on water migration into resin-modified glass ionomer cements: a magnetic resonance micro-imaging study. AB - Magnetic resonance micro-imaging was applied to study water diffusion into resin modified glass ionomer cement restoration and to evaluate the effect of surface coating over restoration. Two cavities were prepared on the labial surface of extracted teeth and restored with resin-modified glass ionomer cement; one was protected with surface coating and the other was not. Immediately after restoration, the teeth were immersed in water. Progress of water diffusion into restorations was monitored by T(1) weighted spin-echo MRI at one-day intervals after the start of immersion. To quantify the water diffusion, a model was developed and compared with imaging data. Best fit yielded an effective water diffusion coefficient D = (2.3 +/- 0.4) 10(-12) m(2)/sec. Experimental results demonstrated that surface coating protects the dental cement against water intrusion from the surface of the restoration which faces the oral cavity. Such coating, however, does not prevent water penetration from the dentine side. PMID- 11064403 TI - SAR and tissue heating with a clinical (31)P MRS protocol using surface coils, adiabatic pulses, and proton-decoupling. AB - In MRS studies using surface transmit coils, accurate assessment of local SAR and RF heating represents a difficult problem involving the coil geometry and electromagnetic and geometric tissue properties. Methodologies to determine the optimum operating parameters for dual-resonant surface coil measurements are presented, based on a standardized coil and protocol used in a multicenter (31)P MRS clinical trial, using adiabatic pulses and bilevel proton decoupling. Spatial distributions of absorbed radiation in human calf and in a tissue-equivalent gel phantom were modeled using finite-element simulations and realistic conductivity and permittivity values. Local SAR in worst-case 1 cm(3) volumes of interest (VOIs) in calf is predicted to be below international guidelines, and the temperature at the skin surface was found to increase due to the RF by less than 2 degrees C and remain below 37 degrees C. The heating rate and maximum temperature in the gel, at positions guided by the simulations, were within guideline values for both extremities and trunk and in reasonable agreement with that predicted. PMID- 11064404 TI - Resting brain metabolic activity in a 4 tesla magnetic field. AB - MRI is a major tool for mapping brain function; thus it is important to assess potential effects on brain neuronal activity attributable to the requisite static magnetic field. This study used positron emission tomography (PET) and (18)F deoxyglucose ((18)FDG) to measure brain glucose metabolism (a measure of brain function) in 12 subjects while their heads were in a 4 T MRI field during the (18)FDG uptake period. The results were compared with those obtained when the subjects were in the earth's field (PET scanner), and when they were in a simulated MRI environment in the PET instrument that imitated the restricted visual field of the MRI experiment. Whole-brain metabolism, as well as metabolism in occipital cortex and posterior cingulate gyrus, was lower in the real (4 T) and simulated (0 T) MRI environments compared with the PET. This suggests that the metabolic differences are due mainly to the visual field differences characteristic of the MRI and PET instruments. We conclude that a static magnetic field of 4 T does not in itself affect this fairly sensitive measure of brain activity. PMID- 11064405 TI - Acute changes in MRI diffusion, perfusion, T(1), and T(2) in a rat model of oligemia produced by partial occlusion of the middle cerebral artery. AB - Oligemic regions, in which the cerebral blood flow is reduced without impaired energy metabolism, have the potential to evolve toward infarction and remain a target for therapy. The aim of this study was to investigate this oligemic region using various MRI parameters in a rat model of focal oligemia. This model has been designed specifically for remote-controlled occlusion from outside an MRI scanner. Wistar rats underwent remote partial MCAO using an undersize 0.2 mm nylon monofilament with a bullet-shaped tip. Cerebral blood flow (CBF(ASL)), using an arterial spin labeling technique, the apparent diffusion coefficient of water (ADC), and the relaxation times T(1) and T(2) were acquired using an 8.5 T vertical magnet. Following occlusion there was a decrease in CBF(ASL) to 35 +/- 5% of baseline throughout the middle cerebral artery territory. During the entire period of the study there were no observed changes in the ADC. On occlusion, T(2) rapidly decreased in both cortex and basal ganglia and then normalized to the preocclusion values. T(1) values rapidly increased (within approximately 7 min) on occlusion. In conclusion, this study demonstrates the feasibility of partially occluding the middle cerebral artery to produce a large area of oligemia within the MRI scanner. In this region of oligemic flow we detect a rapid increase in T(1) and decrease in T(2). These changes occur before the onset of vasogenic edema. We attribute the acute change in T(2) to increased amounts of deoxyhemoglobin; the mechanisms underlying the change in T(1) require further investigation. PMID- 11064406 TI - Displacement imaging of spinal cord using q-space diffusion-weighted MRI. AB - Displacement MR images of water in in vitro rat spinal cord were computed from q space analysis of high b value diffusion-weighted MRI data. It is demonstrated that q-space analysis of heavily diffusion-weighted MRI (qs-DWI) provides MR images in which physical parameters of the tissues such as the mean displacement and the probability for zero displacement of the water molecules are used as contrasts. It is shown that these MR images provide structural information surpassing the spatial resolution of conventional MRI by several orders of magnitude. This imaging methodology was used to follow spinal cord maturation in the rat. It was found that changes in the diffusion characteristics of white matter upon maturation are responsible for the emergence of gray/white matter contrast. The mean displacement of water molecules in the white and gray matter of the mature rat spinal cord was found to be 2-3, and 8-10 microns, respectively. The potential and the limitations of this new imaging methodology for early detection of white matter disorders are discussed. PMID- 11064407 TI - Multidimensional MR mapping of multiple components of velocity and acceleration by fourier phase encoding with a small number of encoding steps. AB - Previous studies have shown that the multi-step approach of velocity or acceleration encoding is highly efficient in terms of the signal-to-noise ratio per unit time. This work describes a multidimensional extension of this method for simultaneously measuring multiple components of velocity and acceleration with a few encoding steps. N flow dimensions were encoded with an ND-matrix, obtained by combining the various flow-encoding gradients. The small matrix obtained with as few as two encoding steps can be extended by zero-filling in all N dimensions and using ND-Fourier transformation to obtain the maximum of the resulting peak in the ND-matrix, which gives simultaneously all the components of velocity and/or acceleration. The processing time was shortened by using a method of phase computation that gives the same precision as Fourier transformation, but is much faster. A rotating disk was used to show that the velocity-to-noise ratio increases with the number of dimensions acquired, demonstrating the efficiency of multidimensional flow measurements. The feasibility of the method is illustrated by 3D maps of the myocardium velocity, and 2D measurement of velocity and acceleration in the ascending aorta-both obtained by multidimensional phase encoding in volunteers. PMID- 11064408 TI - Rapid isotropic diffusion mapping without susceptibility artifacts: whole brain studies using diffusion-weighted single-shot STEAM MR imaging. AB - A subsecond magnetic resonance imaging (MRI) technique for isotropic diffusion mapping is described which, in contrast to echo-planar imaging (EPI), is insensitive to resonance offsets, i.e., tissue susceptibility differences, magnetic field inhomogeneities, and chemical shifts. It combines a diffusion weighted (DW) spin-echo preparation period and a high-speed stimulated echo acquisition mode (STEAM) MRI sequence and yields single-shot images within measuring times of 559 msec (80 echoes). Here, diffusion encoding involved one scan without DW, three DW scans with b = 490 sec mm(-2), and three DW scans with b = 1000 sec mm(-2) (orthogonal gradient orientations). An automated on-line evaluation resulted in isotropic DW images as well as ADC maps (trace of the diffusion tensor). Experiments at 2.0 T covered the brain of healthy subjects in 20 contiguous sections of 6 mm thickness and 2.0 x 2.0 mm(2) in-plane resolution within a total measuring time of 78 sec. High-resolution studies at 1.0 x 1.0 mm(2) (interpolated from 2.0 x 1.0 mm(2) acquisitions) were obtained within 5 min 13 sec using four averages. In comparison with EPI, DW single-shot STEAM MRI exhibits only about half the SNR, but completely avoids regional signal losses, high intensity artifacts, and geometric distortions. PMID- 11064409 TI - Burst excitation for quantitative diffusion imaging with multiple b-values. AB - A quantitative imaging sequence has been developed to exploit the intrinsic sensitivity of Burst NMR data to molecular diffusion. In the scan time of a single spin echo experiment, it is possible to acquire many images of the same slice, with a different T(2) and diffusion weighting. Under favorable conditions, it is possible to obtain both the diffusion coefficient and T(2) from the same experiment; or, by correcting for T(2) relaxation using a control image, more precise diffusion coefficients may be measured. The quantitative values in rat brain are in agreement with those from conventional experiments. The major gains of this method are the potentially reduced scan time, the higher number of acquired images corresponding to different diffusion weightings, the reduced sensitivity to inter-scan motion artifact and to local variations in magnetic susceptibility, and an automatic co-registration between T(2) and diffusion images. Problems with the sequence include a lower signal-to-noise ratio than is achievable with diffusion-weighted spin-echo imaging, the limitation of measuring only in-plane components of diffusion and, at present, single-slice acquisition. PMID- 11064410 TI - Wavelet transform-based Wiener filtering of event-related fMRI data. AB - The advent of event-related functional magnetic resonance imaging (fMRI) has resulted in many exciting studies that have exploited its unique capability. However, the utility of event-related fMRI is still limited by several technical difficulties. One significant limitation in event-related fMRI is the low signal to-noise ratio (SNR). In this work, a method based on Wiener filtering in the wavelet domain is developed and demonstrated for denoising event-related fMRI data. Application of the technique to simulated and experimental data demonstrates that the technique is effective in reducing noise while preserving neuronal activity-induced response. PMID- 11064411 TI - B(0)-fluctuation-induced temporal variation in EPI image series due to the disturbance of steady-state free precession. AB - Steady-state free precession (SSFP) can develop under a train of RF pulses, given the condition TR < T(2). SSFP in multi-shot imaging sequences has been well studied. It is shown here that serial single-shot echo-planar imaging (EPI) acquisition can also develop SSFP, and the SSFP can be disturbed by B(0) fluctuation, causing voxel-wise temporal variation. This SSFP disturbance is predominantly present in cerebrospinal fluid (CSF) regions due to the long T(2) value. By applying a sufficiently strong crusher gradient in the EPI pulse sequence, the temporal variation induced by SSFP disturbance can be suppressed due to diffusion. Evidence is provided to indicate that physiological motions such as cardiac pulsation and respiration could affect the voxel-wise time courses through the mechanism of SSFP disturbance. It is advised that if the disturbance is observed in serial EPI images, the crusher should be made stronger to eliminate the unwanted temporal variation. PMID- 11064412 TI - Applying the uniform resampling (URS) algorithm to a lissajous trajectory: fast image reconstruction with optimal gridding. AB - Various kinds of nonrectilinear Cartesian k-space trajectories have been studied, such as spiral, circular, and rosette trajectories. Although the nonrectilinear Cartesian sampling techniques generally have the advantage of fast data acquisition, the gridding process prior to 2D-FFT image reconstruction usually requires a number of additional calculations, thus necessitating an increase in the computation time. Further, the reconstructed image often exhibits artifacts resulting from both the k-space sampling pattern and the gridding procedure. To date, it has been demonstrated in only a few studies that the special geometric sampling patterns of certain specific trajectories facilitate fast image reconstruction. In other words, the inherent link among the trajectory, the sampling scheme, and the associated complexity of the regridding/reconstruction process has been investigated to only a limited extent. In this study, it is demonstrated that a Lissajous trajectory has the special geometric characteristics necessary for rapid reconstruction of nonrectilinear Cartesian k space trajectories with constant sampling time intervals. Because of the applicability of a uniform resampling (URS) algorithm, a high-quality reconstructed image is obtained in a short reconstruction time when compared to other gridding algorithms. PMID- 11064413 TI - Analytic calculations of the E-fields induced by time-varying magnetic fields generated by cylindrical gradient coils. AB - Analytic expressions which allow the direct calculation of the electric field generated inside an infinite conducting cylinder by varying the current through the wires of any cylindrical coil are presented. These expressions provide some general insight into the spatial characteristics of the electric field generated inside the body by switched gradients and can be used to evaluate the locations where nerve stimulation by rapid gradient switching is likely to occur. They may also be employed at the design stage to produce gradient coils which can provide higher gradient switching rates without causing nerve stimulation. Using these expressions the electric field patterns produced by transverse and longitudinal, whole-body gradient coils were calculated. Example data are presented along with the associated magnetic field patterns. The effect on the induced electric field pattern of varying the body size and the size of the region of gradient linearity was explored. PMID- 11064414 TI - Electroencephalography during functional echo-planar imaging: detection of epileptic spikes using post-processing methods. AB - EEG has been used to trigger functional MRI of patients with focal epilepsy, but EEG can be obscured by artifacts during MR data acquisition, and no continuous correlation of EEG and MRI has been possible without limiting the image time. Artifacts caused by an MRI sequence were investigated in five healthy subjects, and an EEG of five patients with epileptic discharges was recorded during echo planar imaging. All interfering frequencies in the EEG were discrete and defined by loop structures in the MRI sequence. In post-processing of the EEG interfering frequencies were automatically detected by comparing the frequency spectra of the EEG recorded before and during imaging. After elimination of interfering frequencies by filters in the time domain or by Fourier transform, reliable spike detection in the EEG recorded during MR data acquisition became feasible, without loss of EEG quality. PMID- 11064415 TI - Determination of pH using water protons and chemical exchange dependent saturation transfer (CEST). AB - Solution pH was measured using water proton NMR via chemical exchange dependent saturation transfer (CEST) with selected chemical exchange sites. Several useful pH-sensitive proton chemical exchange agents were found: 5,6-dihydrouracil, 5 hydroxytryptophan, and a combination of 5-hydroxytryptophan and 2 imidazolidinethione. A ratiometric approach was developed that permitted pH determinations that were independent of water T(1) or exchange site concentration. PMID- 11064416 TI - Effect of NC100150 injection on the (1)H NMR linewidth of human whole blood ex vivo: dependency on blood oxygen tension. AB - The linewidth of the (1)H NMR signal (7.05 T) of human whole blood titrated with a superparamagnetic contrast agent (NC100150 injection) was evaluated at different blood oxygen tensions. In deoxygenated blood and low contrast agent concentrations, NC100150 injection caused a decrease in linewidth. After reaching a minimum, the linewidth increased as the concentration of NC100150 injection increased. At the concentration corresponding to the minimum linewidth, the magnetization of the extracellular space containing the NC100150 injection was equal to that of the paramagnetic (deoxygenated hemoglobin) intracellular space. The minimum linewidth is therefore consistent with a complete elimination of the local microscopic susceptibility effect, the major cause of linebroadening. Additionally, phantom studies were performed at 1.5 T, confirming that the contrast enhancement of NC100150 injection in blood is dependent on oxygen tension. The data suggest that NC100150 injection may be useful in differentiating vessels with varying relative oxygen tensions. PMID- 11064417 TI - Noninvasive pulmonary perfusion imaging by STAR-HASTE sequence. AB - The STAR-HASTE sequence has been shown to be useful for perfusion imaging in areas that are plagued by magnetic susceptibility artifacts. Pulmonary perfusion imaging with this technique was attempted in this study. Quantitative analysis was also conducted, using an appropriate kinetic model in one subject. In six healthy subjects, gradual enhancement was observed in pulmonary artery to distal lung parenchyma when inflow time was increased. Our initial results suggest that noninvasive evaluation of pulmonary perfusion by magnetic resonance imaging without administration of an exogenous agent is possible. PMID- 11064418 TI - MR virtual colonography using hyperpolarized (3)He as an endoluminal contrast agent: demonstration of feasibility. AB - Hyperpolarized gas MR virtual colonography was performed in plastic phantoms and in the dog colon. (3)He was laser polarized in a prototype commercial system. 2D and 3D gradient echo sequences were used to image the noble gas-filled structures. The hyperpolarized (3)He within the plastic tube and colon lumen produced high signal, providing excellent contrast from the surrounding structures. The virtual colonoscopic analysis of the canine dataset allowed visualization of the colonic features and the colonic wall from inside the colon. (3)He colonoscopy is a novel technique to visualize the colon with MRI with the application of an inert gaseous endoluminal contrast agent. PMID- 11064419 TI - Method for rapidly determining and reconstructing the peak arterial frame from a time-resolved CE-MRA exam. AB - A method that determines the information necessary to reconstruct a single vascular image from a time-resolved CE-MRA exam is presented. Raw k-space data are used to approximate the time course of the contrast passage prior to image reconstruction. The resulting k-space contrast curve is used to select the data corresponding to peak arterial enhancement. These data are reconstructed and immediately presented for physician review, with the entire time-series of images available at a later time for more detailed diagnosis. This approach dramatically reduces the latency between acquisition of large 4D (3D plus time) data sets and presentation of a diagnostic quality time frame. This algorithm has proven successful in the imaging of several anatomical regions and-in exams that do not require a breath hold-permits the use of an acquisition method that produces a contrast-enhanced angiogram without a timing scan. PMID- 11064420 TI - Evolution and development at the National Science Foundation, U.S.A. PMID- 11064421 TI - Zebrafish nma is involved in TGFbeta family signaling. AB - Bone morphogenetic proteins (BMP) are members of the TGFbeta superfamily of secreted factors with important regulatory functions during embryogenesis. We have isolated the zebrafish gene, nma, that encodes a protein with high sequence similarity to human NMA and Xenopus Bambi. It is also similar to TGFbeta type I serine/theronine kinase receptors in the extracellular ligand-binding domain but lacks a cytoplasmic kinase domain. During development, nma expression is similar to that of bmp2b and bmp4, and analysis in the dorsalized and ventralized zebrafish mutants swirl and chordino indicates that nma is regulated by BMP signaling. Overexpression of nma during zebrafish and Xenopus development resulted in phenotypes that appear to be based on inhibition of BMP signaling. Biochemically, NMA can associate with TGFbeta type II receptors and bind to TGFbeta ligand. We propose that nma is a BMP-regulated gene whose function is to attenuate BMP signaling during development through interactions with type II receptors and ligands. PMID- 11064422 TI - Functional interaction of vega2 and goosecoid homeobox genes in zebrafish. AB - The gastrula organizer forms in the dorsal region of the zebrafish embryo, where the bozozok/dharma homeobox gene downregulates expression of the vega1 transcriptional repressor. Here, we describe a novel Vega family homeobox gene, vega2. Expression of vega2 is initiated at the ventral blastoderm margin during blastula stages, and by gastrulation becomes complementary to but partially overlapping with the dorsal expression domain of the homeobox gene goosecoid (gsc). This dorsal exclusion of vega2 expression is not observed in bozozok mutants in which organizer formation is impaired. Both vega2 and vega1 can physically interact with Gsc. Zebrafish embryos injected with vega2 mRNA failed to express gsc and developed a headless phenotype. Conversely, a putative dominant negative form of vega2, VP16-vega2, elicited the expansion of gsc expression and a dorsalized phenotype. We suggest that vega2, in cooperation with vega1, functions as a negative regulator of organizer genes including gsc, and participates in the refinement of the gastrula organizer domain. PMID- 11064423 TI - Mosaic analysis of GL2 gene expression and cell layer autonomy during the specification of Arabidopsis leaf trichomes. AB - Homozygous glabra2 (gl2) mutant Arabidopsis thaliana Landsberg erecta plants with only a few rudimentary single spiked trichomes on the leaf margin were transformed with a genomic clone of GL2, resulting in partial restoration of the normal leaf trichome phenotype. The introduced GL2 transgene was configured as part of an FLP recombinase-responsive gene switch, which permitted visibly marked gl2 mutant clonal sectors to be generated by FLP recombinase-mediated deletion of the GL2 transgene with concomitant activation of a previously silent beta glucuronidase (GUS) marker gene. GUS marked sectors extending through all three leaf cell layers (L1, L2, and L3) displayed the anticipated gl2 mutant phenotype, whereas immediately adjacent unmarked tissue, and unmarked tissues overlaying GUS sectors restricted to the L2 and/or L3 cell layers, retained the GL2 restored phenotype. These data support the view that the GL2 gene product acts in a region autonomous manner within a single cell layer and indicate that GL2 gene expression in the L1 layer is sufficient for GL2-directed outgrowth of trichomes. PMID- 11064424 TI - Universal GFP reporter for the study of vascular development. AB - We report the generation and characterization of transgenic mouse and zebrafish expressing green fluorescent protein (GFP) specifically in vascular endothelial cells in a relatively uniform fashion. These reporter lines exhibit fluorescent vessels in developing embryos and throughout adulthood, allowing visualization of the general vascular patterns with single cell resolution. Furthermore, we show the ability to purify endothelial cells from whole embryos and adult organs by a single step fluorescence activated cell sorting. We expect that these transgenic reporters will be useful tools for imaging vascular morphogenesis, global gene expression profile analysis of endothelial cells, and high throughput screening for vascular mutations. PMID- 11064425 TI - Cell size and the morphogenesis of wing hairs in Drosophila. AB - Almost all epidermal cells on the Drosophila wing produce a single cuticular hair. This is formed in the pupae from a microvillus-like cell projection called the prehair. Previous experiments have shown the existence of two mechanisms that ensure that only a single hair is made. One is the restriction of prehair initiation to a small subregion of the cell by the action of the frizzled tissue polarity pathway. The second is a system that ensures the integrity of the prehair. Mutations and drugs that inhibit the actin cytoskeleton lead to the splitting of a single prehair into multiple smaller hairs. We report that large polyploid cells produce multiple hairs both because they form multiple independent prehair initiation centers and because the larger than normal hairs these cells produce have a tendency to split. We show that reducing cell size by starvation partially suppresses the phenotype seen in polyploid cells and that increasing apical cell surface area by mechanical stretching also results in the formation of multiple prehair initiation centers. We also show that the frizzled tissue polarity pathway is functional in large polyploid cells even if it is unable to restrict prehair initiation to a small region of the cell. We conclude that both of these cellular systems are limited in their ability to scale to accommodate larger cell size. PMID- 11064426 TI - Respiratory pacemaker cells responsive to CO(2) in the upper medulla: dose response and effects of mediators. AB - We previously reported on the presence of respiratory pacemaker cells that are highly sensitive to CO(2), in a region of the medulla oblongata in the fetal rat, 2 mm rostral to the obex. We now report on the CO(2) dose responses of these cells, as well as their responsiveness to certain chemical agents known to affect breathing in the fetus. Twenty-day-old fetal Sprague Dawley rats were block dissected, and the cells of target areas were dissociated as previously described. Neuronal cells were plated on a medullary background and placed in the incubator with 10% CO(2) for 2-3 weeks. Cells were then studied using patch-clamp techniques. Pacemaker cells with single or bursting potentials showed responsiveness to CO(2) starting with pulses of 10 msec. Irregular beating or silent cells had poor or absent responsiveness to CO(2). Pacemaker cells responded to norepinephrine with increased firing potential; this action was blocked by metropolol. PGE(2) had no effect on pacemaker-cell activity, but indomethacin increased the spike frequency from 336+/-41 to 384+/- 65 spikes/min. Morphine stimulated the pacemaker cells from 205+/-25 to 272+/-29 spikes/min; this was blocked by naloxone. Finally, a placental extract, which inhibited breathing in the unanesthetized fetal sheep preparation, increased the activity of pacemaker cells from 301+/-35 to 452+/-52 spikes/min. In all of the above, irregular beating cells responded poorly and silent cells did not respond. The findings indicate that these pacemaker cells are uniquely designed to respond to CO(2) and have some properties which allow them to respond to certain chemical mediators in a manner similar to that of the whole respiratory system in vivo. PMID- 11064427 TI - Aerosol therapy in cystic fibrosis: a survey of 54 CF centers. AB - Aerosol therapy has become increasingly important in the treatment of lung disease of patients with cystic fibrosis (CF). Still, many questions concerning this therapy remain unanswered. It is unclear at what age aerosol therapy should be started; which aerosolized drugs are essential in the treatment of CF lung disease; which delivery system(s) should be used; and how aerosol therapy should be timed in relation to physiotherapy. We hypothesized that large differences in aerosol treatment practices between CF centers would be present. To investigate this, we performed an observational survey to evaluate different aspects of aerosol therapy. A questionnaire was sent to 102 CF centers in 28 different countries. A completed questionnaire was returned by 54 out of 94 centers (57%). In these 54 centers, 7,324 CF patients were treated. Substantial differences were found in aerosol therapy between centers. Patients below age 1 year were not treated with any form of aerosol therapy in 10% of the centers, while 37.5% of the centers treated all of these patients. The timing of nebulization and physiotherapy varied substantially for many important and expensive drugs. We conclude that many aspects of aerosol therapy in cystic fibrosis need to be executed in a more rational and evidence-based manner than is currently the case. PMID- 11064428 TI - Daycare attendance before the age of two protects against atopy in preschool age children. AB - Early attendance at daycare has been shown to protect against atopy, as defined by a positive skin prick test. One proposed hypothesis for this association is that early exposure to other children protects against atopy by facilitating the spread of infections among children. An alternative hypothesis is that children attending daycare centers have less atopy due to lower levels of exposure to indoor allergens. Our aim was to determine whether attendance at daycare before age 2 years protects against atopy in Australian preschool age children and to test the two alternative hypotheses, as well as a number of potential confounding factors. We conducted a cross-sectional study of children aged 3-5 years living in one humid coastal city (Lismore, n = 286) and one dry, inland city (Wagga Wagga, n = 364) in New South Wales, Australia, in 1995. Atopy was assessed by skin prick tests to six common allergens. Daycare attendance and other possible risk factors for atopy were measured by a parent-completed questionnaire. Children who attended daycare before age 2 years had a reduced risk of atopy at 3 5 years. The greatest effect was seen in children who attended a daycare center (odds ratio (OR), 0.26; 95% CI, 0.14-0.50) rather than family daycare (OR, 0.66; 95% CI, 0.41-1.04). The results of this study do not support either of the proposed hypotheses, nor can the effect be explained by any of the other potential confounders measured. Further work is needed to determine the exposure that is responsible for the protective effect of daycare attendance on atopy. PMID- 11064429 TI - Correlation between reversibility of airway obstruction and exhaled nitric oxide levels in children with stable bronchial asthma. AB - Recent trials measuring exhaled nitric oxide (eNO) concentrations have suggested that it may be a useful measure of ongoing airway inflammation in patients with asthma. The purpose of this study was to examine the relationship between eNO levels and baseline as well as postbronchodilator spirometry, a measurement commonly used in the clinical setting to determine the severity of asthma and as a guide to therapeutic decisions. Forty-nine patients between the ages of 5-16 years with physician-diagnosed asthma who attended the pediatric pulmonary clinic for a routine asthma visit with spirometric evaluation were recruited for the study. eNO levels prior to spirometry were obtained before and after receiving inhaled beta(2) agonist. eNO samples were collected in impermeable bags (Tedlar) and assayed within 24 hr by chemiluminescence. Regression analysis was used to assess the relationships between pre- and postbronchodilator eNO and spirometric variables. eNO was also compared in patients receiving and not receiving inhaled corticosteroids (ICS), as well as those whose therapy had been increased after evaluation by a pediatric pulmonologist or allergist. We found no significant difference between the levels of eNO before and after inhalation of beta(2) agonist (P = 0.60 paired t-test). Positive correlation was found between eNO vs. percentage change in FEV(1) (r = 0.35, P = 0.01) and percentage change in FEF(25 75% )(r = 0.29, P = 0.04). A negative correlation was found between prebronchodilator FEV(1) and eNO (r = -0.29, P = 0.03). Patients on ICS had lower mean eNO levels (29.9 vs. 47.6 parts per billion (ppb), P = 0.053) than those not receiving ICS. Patients whose ICS therapy was increased had higher mean eNO levels (47.2 vs. 26.9 ppb, P = 0.018) than those not having ICS therapy increased. We suggest that eNO levels could be a clinically useful measurement of asthma severity and could be used as an adjunct to spirometry to determine appropriate treatment plans. Longitudinal clinical trials are needed to determine if eNO can enhance therapeutic decisions for asthmatic children. PMID- 11064430 TI - Pulmonary complications after bone marrow transplantation in children: twenty four years of experience in a single pediatric center. AB - In children, pulmonary sequelae contribute to early and late morbidity after bone marrow transplantation (BMT). Between 1975-1999, we performed 152 BMTs in 138 pediatric patients with malignant and nonmalignant diseases. Allogenic bone marrow was used from 99 HLA identical siblings and from 23 other related or unrelated donors. Autologous marrow was used in 30 transplantations. Median age was 8. 6 years (range, 1.1-22.4) at time of BMT. The median survival was 42%, the survival time was 6.5 years (range, 0.8-23.1), and the median follow-up time was 6.8 years (range, 0.8-23.2). Seventeen patients had severe respiratory complications. Early severe respiratory complications leading to death within the first 4 months after BMT were due to pulmonary edema (n = 1), or fungal (n = 3), bacterial (n = 1), or viral (n = 2) pneumonia. Late severe respiratory sequelae were defined as persistent respiratory symptoms for more than 4 months despite treatment, and these occurred in 10 patients, of whom 5 died. Underlying diagnoses covered a wide spectrum, including bronchiolitis obliterans (n = 3), severe restrictive lung disease (n = 2), idiopathic pneumonia syndrome (n = 3), chronic bronchitis (n = 1), and hepatopulmonary syndrome (n = 1). The overall probability for death was 0.58, and for death from severe respiratory complications, 0.16. With improved HLA matching, fewer BMTs after relapsed or primary progressive disease, and improved supportive care, including the usage of CMV negative blood products, after 1990 the probability of death from severe respiratory complications was only 0.04, whereas before 1990 it was 0.23 (P = 0.029; in each time period, n = 69). The disease spectrum has changed from initially more infectious complications to bronchiolitis obliterans and idiopathic pneumonia syndrome. Lung function measurements performed in 85 of 138 patients usually showed a mild restrictive pattern. To identify those children as early as possible who are at risk for severe respiratory complications, a close longitudinal follow-up after BMT by pediatric pulmonologists is necessary. PMID- 11064431 TI - Perinatal Ureaplasma urealyticum infection increases the need for hospital treatment during the first year of life in preterm infants. AB - To explore the association of perinatal Ureaplasma urealyticum infection and the need for hospital care during infancy, a cohort of preterm infants were prospectively followed for 12 months. Perinatal U. urealyticum infection was defined as the presence of U. urealyticum in the samples obtained from the trachea and blood. During the first year of life, the infants of the study cohort required 73 hospital admissions resulting in 734 hospital days. The 22 infants with perinatal U. urealyticum infection needed more hospital days for therapy than the 18 infants without infection (546 vs. 188 days, P = 0.042). The difference was caused by an increase in respiratory tract diseases among children with perinatal U. urealyticum infection. Chronic lung disease caused more admissions in infants with perinatal U. Urealyticum infection than without it (P = 0.035). The results indicate that perinatal U. urealyticum infection affects the health of premature infants far beyond the perinatal period. PMID- 11064432 TI - Pulse oximetry: what's normal in the newborn nursery? AB - The objective of this study was to establish normal values for pulse oximetry saturation (POS) in healthy newborn infants in the nursery. POS values were obtained from the right (R) hand and R foot at admission, 24 hr, and at discharge. The following information was recorded: postnatal age, activity state, gender, gestational age (GA), birth weight (BW), mode of delivery (MOD), and Apgar scores. Charts were reviewed and follow-up information was obtained for newborns with measurements < or =92%. The study group consisted of a convenience sample of newborn infants, excluding those on supplemental oxygen. Seven hundred eighteen patients were studied: 51% males, 28% cesarean sections, gestational age 39.3+/-1.6 weeks (mean +/- SD), birth weight 3370+/-550 g, and median Apgar scores 8 and 9. The mean POS was 97.2 +/-1.6%, and the median value was 97%. Only postnatal age and activity state affected POS significantly. POS increased 0.17% per 24 hr in the nursery (P = 0. 0001). POS values obtained while the infants were fussy and crying were lower compared to measurements obtained while sleeping [mean decreases: 0.44% while fussy (P = 0.001), 0.98% while crying (P = 0.0001)]. We conclude that newborns in the nursery have an overall mean POS of 97.2% (+/-2 SD: 94-100%). Mean POS values increase to a small degree with increasing postnatal age. Fussy and crying newborns have lower POS values compared to quiet and sleeping newborns. These reference data can be used in the evaluation of POS measurements in symptomatic newborn infants. PMID- 11064433 TI - Mouse models of chronic lung infection with Pseudomonas aeruginosa: models for the study of cystic fibrosis. AB - The discovery of the CFTR gene in 1989 has lead to rapid progress in understanding the molecular basis of cystic fibrosis (CF) and the biological properties of the cystic fibrosis transmembrane conductance regulator (CFTR) protein. However, more than 10 years later, recurrent lung infections with Pseudomonas aeruginosa, which lead to chronic lung disease and eventual respiratory failure, remain the major cause of morbidity and mortality among CF patients. A distinguishing feature of lung disease in CF is an exaggerated and persistent inflammatory response, characterized by the accumulation of excessive numbers of neutrophils and dysregulated cytokine production. The events leading to the establishment of lung infection with P. aeruginosa, especially the inflammatory and immunological events, and the relation between the CF defect and infection, remain largely undefined. Progress in this area has been hampered by the lack of a suitable animal model. An exciting achievement in the past few years has been the development of a number of variants of CFTR-deficient mice which exhibit defective cAMP-mediated Cl(-) conductance and have a range of clinical phenotypes from mild to severe. In parallel, a model of chronic P. aeruginosa lung infection has been established in genetically and immunologically well-defined inbred mouse strains which differ in susceptibility to this infection in the lung. BALB/c mice are resistant, while DBA/2 mice are extremely susceptible, with high mortality within 3 days of infection. C57BL/6 and A/J mice are relatively susceptible and experience low mortality. Furthermore, the bacterial load correlates with the magnitude and quality of the inflammatory response in the infected lungs of BALB/c and C57BL/6 mice. Although results of infection studies in CFTR-deficient mice have been variable, C57BL/6 Cftr(m1UNC)/Cftr(m1UNC) knockout mice compared to littermate control mice are highly susceptible to chronic P. aeruginosa infection in the lung. The availability of CFTR knockout mice and non-CF inbred mice differing in susceptibility to chronic P. aeruginosa infection offers useful tools for progress in understanding the genesis of chronic P. aeruginosa infection and the ensuing inflammation in the CF lung, as well as the relation between the CF defect and infection. Information generated from these studies will provide the rationale for the development of novel immunomodulatory measures capable of ameliorating or modulating the chronic inflammation associated with CF lung disease. PMID- 11064434 TI - Granular cell tumor of the bronchus. AB - Persistent atelectasis and recurrent pneumonia in the same location should raise suspicion of congenital anomalies or obstructing lesions of the bronchus leading to the affected area. We present an 8-year-old black female with a history of recurrent fever, cough, atelectasis of the right middle and lower lobes, and weight loss for several months. Flexible bronchoscopy revealed a polypoid mass obstructing the bronchus intermedius. Biopsy of the neoplasm demonstrated a granular cell tumor (GCT). The patient had a lobectomy of the right lower and middle lobes. She had no recurrence of the tumor after several years of follow up. PMID- 11064435 TI - Morgagni hernia: an unexpected cause of respiratory complaints and a chest mass. AB - Morgagni hernia (MH) is the least common type of congenital diaphragmatic hernias. Although its course is often asymptomatic, it may be associated with various respiratory and gastrointestinal symptoms. We describe 7 children with MH during a 5-year period in three pediatric centers in Turkey. All children had acute or chronic respiratory symptoms; cough was the most frequent. The diagnosis was made by posterior-anterior (PA) and lateral chest X-rays. The PA chest X-rays showed a homogenous mass in 2 and a gas-filled cystic image in 3 children in the right cardiophrenic angle. A retrocardiac homogeneous density in one child, and bilateral consolidation in lower lung areas in another child were also seen. All lateral chest X-rays showed gas-filled bowel loops above the diaphragm. The diagnosis was confirmed by barium-contrast radiograph. Four patients had five additional anomalies, i.e., ventricular septal defect, right inguinal hernia, congenital hip dislocation, pectus carinatum, and obstruction of the uretero pelvic junction. All of the hernias were repaired by an abdominal approach. There were no complications or recurrences during follow-up. In conclusion, MH should be considered in the differential diagnosis of cases of long-standing respiratory symptoms and/or when an unexplained radiological image, especially on the right cardiophrenic area, is present. PMID- 11064436 TI - Selected abstracts PMID- 11064437 TI - Pattern of cytokine secretion by peripheral blood cells of patients with multiple sclerosis in Brazil. AB - Autoimmune T cells play a key role as regulators and effectors of organ-specific autoimmune disease. In multiple sclerosis (MS), activated T cells specific for myelin components produce a plethora of inflammatory cytokines and mediators that contribute to myelin damage. The production of proinflammatory and regulatory cytokines by peripheral blood cells from patients with active and stable MS and healthy controls were examined. The results show that TNF alpha production was somewhat elevated in active MS with no significant increase in the level IFN gamma, whereas in the chronic phase the anti-inflammatory cytokines IL-10 and TGF beta increased, accompanied by a reduction in IFN gamma when stimulated by myelin basic protein. Multiple Sclerosis (2000) 6 293 - 299 PMID- 11064438 TI - Kinetic profiles of cerebrospinal fluid anti-MBP in response to intravenous MBP synthetic peptide DENP(85)VVHFFKNIVTP(96)RT in multiple sclerosis patients. AB - Multiple sclerosis [MS], a demyelinating disease of the central nervous system associated with inflammation and gliosis, may be an autoimmune disease with T lymphocytes and autoantibodies to myelin protein(s). This study deals exclusively with B cell autoimmunity to myelin basic protein (MBP). T lymphocytes and anti MBP share a common MBP epitope located between P(85) and P(96) which contains the essential contact residues H(88)FFK(91) for the trimolecular complex. The purpose of this Phase I open label clinical study was to monitor CSF anti-MBP in patients with chronic progressive MS subsequent to IV administration of synthetic peptide (sp) MBP82-98 namely DEN(85)VVHFFKNIVTP(96)RT. Fifty-six patients who participated in this project were assigned to two groups: a 'control group' of 15 patients who received IV saline injections every 6 months for the first 2 years of the study and a 'peptide group' of 41 patients who received IV spMBP82-98 from the beginning of the study and then infrequently subsequent to a rise of their CSF anti-MBP. In the control group antibody levels remained persistently elevated during the 2 year period. Patients in the 'peptide group' segregated into four kinetic profiles: Cohort A (15 patients) illustrated prolonged anti-BMP suppression into the normal range. Cohort B (10 patients) illustrated significant anti-MBP suppression into the normal range for shorter durations. Cohort C (eight patients) showed significant CSF anti-MBP suppression after the initial injection but lost the ability to suppress the autoantibody titer following subsequent injections. Cohort D (eight patients) failed to show significant CSF anti-MBP suppression. In conclusion the B cell tolerizing effect of spMBP82-98 segregated into four kinetic profiles; this molecular variability should be considered in attempts to develop specific 'peptide therapies' for the broad range of clinical profiles currently diagnosed as 'multiple sclerosis'. Multiple Sclerosis (2000) 6 300 - 311 PMID- 11064439 TI - Importance of paraclinical and CSF studies in the diagnosis of MS in patients presenting with partial cervical transverse myelopathy and negative cranial MRI. AB - Patients presenting with isolated partial cervical myelopathy are at high risk for development of multiple sclerosis (MS), especially if lesions suggestive of demyelination are present on cranial magnetic resonance imaging (MRI). This risk is lower, though not precisely known, in patients whose cranial MRI is normal. This clinical issue was addressed by examining the role of paraclinical studies in establishing a diagnosis of MS at the time of initial presentation. Twelve consecutive patients, mean age of 32.2 years, seen over 6.5 years were identified prospectively and included in this study. Numbness was the presenting symptom in 11 of these patients. Symptoms completely resolved in nine patients regardless of treatment with glucocorticoids. Evoked potential (EP) and cerebrospinal fluid (CSF) examinations assisted in establishing a diagnosis of laboratory-supported definite (LSDMS) or clinically probable (CPMS) MS in six patients at the time of presentation. During a clinical follow-up period of 4.1 years, four developed recurrent neurologic deficits leading to the establishment of a diagnosis of clinically definite MS (CDMS). The presence of a solitary, non-specific lesion on cranial MRI resulted in an increased risk for the development of definite MS. In patients with a clinically isolated cervical partial transverse myelitis (TM) and normal cranial MRI, an accurate diagnosis of MS can usually be made. Revision of the diagnostic criteria for LSDMS is warranted. Multiple Sclerosis (2000) 6 312 - 316 PMID- 11064440 TI - Histocompatibility antigen (HLA) associations with multiple sclerosis in Iran. AB - Human leukocyte antigen (HLA) types were obtained from 79 Iranian patients with multiple sclerosis and compared with 100 controls. The prevalence of HLA-A24 (30.3% versus 18.0%), HLA-DR2 (43.0% versus 28. 0%) and HLA-DR15 (36.7% versus 23.0%) were significantly increased in multiple sclerosis patients compared with controls. However age at onset, and disease status (relapsing - remitting or primary progressive) did not show an association with any particular HLA type. Multiple Sclerosis (2000) 6 317 - 319 PMID- 11064441 TI - Enhanced magnetic resonance imaging in multiple sclerosis. AB - Gadolinium-enhanced magnetic resonance imaging (MRI) is very sensitive in the detection of active lesions of multiple sclerosis (MS) and has become a valuable tool to monitor the evolution of the disease either natural or modified by treatment. In the past few years, several studies, on the one hand, have assessed several ways to increase the sensitivity of enhanced MRI to disease activity and, on the other, have investigated in vivo the nature and evolution of enhancing lesions using different non-conventional MR techniques to better define the relationship between enhancement and tissue loss in MS. The present review is a summary of these studies whose results are discussed in the context of MS clinical trial planning and monitoring. Multiple Sclerosis (2000) 6 320 - 326 PMID- 11064442 TI - MTR and T1 provide complementary information in MS NAWM, but not in lesions. AB - MTR and T1 relaxation times are abnormal in MS lesions and NAWM, and may reflect tissue damage such as demyelination and axonal loss. Their relationship and potential to provide complementary information in tissue characterisation is explored. The aim of this study was to document the relationship between magnetisation transfer ratio (MTR) and T1 relaxation time in Multiple Sclerosis (MS) lesions and normal appearing white matter (NAWM) in order to determine whether the combination provides a more comprehensive tissue characterisation than either parameter in isolation. Ten patients with relapsing remitting MS and 10 age matched healthy controls underwent imaging using a protocol which included the measurement of both MTR and T1 relaxation times. The MTR and T1 values were compared statistically using a commonly adopted correlation approach and a mixed model regression approach. There was a strong correlation between MTR and T1 in MS lesions (r=0.74). The correlation was seen equally in T1 hypointense and isointense lesions. The relationship was much weaker in MS NAWM (r=0.24) and no correlation was found in control white matter (r=0.06). Mixed-model regression analysis confirmed that the relationship between T1 and MTR is strongly dependent upon tissue type (MS lesion, MS NAWM, or control white matter). The relationship between MTR and T1 relaxation time measurements varies markedly between pathological and normal tissue types. In MS, the complementary information obtained from MTR and T1 is most apparent in NAWM. The results emphasise the potential for combinations of MR parameters to improve tissue characterisation, which in turn should improve understanding of disease pathology and treatment monitoring. Multiple Sclerosis (2000) 6 327 - 331 PMID- 11064443 TI - A longitudinal study of ventricular volume in early relapsing-remitting multiple sclerosis. AB - The specific aim of this study was to determine whether progressive brain atrophy could be detected within 18 months of establishing a diagnosis of relapsing remitting multiple sclerosis (RRMS). Fifteen patients with clinically definite RRMS (mean disease duration from first symptom=6 months, mean EDSS=1.2) completed 6 - 14 monthly quantitative MRI sessions. The volume of the lateral ventricles was determined each month using a semi-automated thresholding technique from T1 weighted axial images. The number of new monthly gadolinium-enhancing (Gd+) lesions and EDSS scores were also recorded. Lateral ventricular volumes increased significantly during this study. When individual data were examined, statistically significant changes were observed in six of 15 patients. Monthly change in ventricular volume was related to baseline EDSS and total number of new Gd(+) lesions. These observations indicate brain atrophy, a putative imaging marker of diffuse demyelination and axonal loss, can occur as early as 18 months after first symptoms of RRMS, and is related to the baseline level of disability and to the number of new Gd+ lesions. Multiple Sclerosis (2000) 6 332 - 337 PMID- 11064444 TI - Quality of life during the first 6 months of interferon-beta treatment in patients with MS. AB - OBJECTIVES: To determine the quality of life (QoL) of MS patients during the initial 6 months of treatment with interferon-beta (IFN-beta). Furthermore, to determine whether changes in QoL relate to disability, emotional state, therapeutic expectations or side effect profile. BACKGROUND: IFN-beta has been shown to have beneficial effects on the course of MS. Since the aim of IFN-beta treatment is not to cure but to slow down the disease it is important to know how this treatment affects QoL. Surprisingly, the impact of treatment with IFN-beta on QoL measures has not been extensively studied so far. METHODS: Case report documentation, including EDSS, SF-36 and MADRAS scores, of 51 relapsing-remitting MS patients treated with IFN-beta was obtained at baseline and at months 1, 3 and 6. Patients also filled in a form about their expectations of therapy and a questionnaire on side effects. RESULTS: During treatment there was a significant linear trend indicating improvement in the role-physical functioning (RPF) scale of the SF-36 (F(1,50)=4.9, P=0.032). A transient decrease at month 1 was found in the scale for bodily pain, indicating more experienced pain (F(1,50)=19.8, P<0.001). Subgroup analysis showed that patients with most depressive symptoms on the MADRAS at baseline contributed most to the increase in RPF scores over time (F(1,24)=5,6 P=0.026). Furthermore, we found associations between adverse event scores and several domains of QoL. CONCLUSIONS: Our findings suggest that IFN beta therapy has an impact on QoL of MS patients in that it improves role physical functioning and transiently worsens experienced bodily pain. QoL during treatment with IFN-beta is influenced by depressive symptoms at baseline as well as by treatment-associated side-effects. Multiple Sclerosis (2000) 6 338 - 342 PMID- 11064445 TI - Multiple sclerosis, disease modifying treatments and depression: a critical methodological review. AB - BACKGROUND: Major depression affects one in two patients with multiple sclerosis (MS) during the course of their lifetime. This adds to the morbidity associated with the disorder and may contribute to an increased mortality rate because of suicide. Over the past few years, with the advent of disease modifying treatments for MS, a new concern with respect to mood has arisen, namely the possibility that some of these drugs may have depression as a clinically significant side effect. OBJECTIVE: To ascertain whether disease modifying treatments in MS are associated with the development of depression or the worsening of a depressive illness. METHODOLOGY: A MEDLINE and PSYCHLIT search focusing on depression and disease modifying treatments going back to 1993 (the publication date of the results of the first randomised, placebo controlled trial). The methodology pertaining to the assessment of depression is critically reviewed. Furthermore, a critical summary is provided of treatment modalities for the depressed MS patient. RESULTS: There are conflicting data that depression may occur with some disease modifying drugs, particularly interferon beta-1b. However, all studies reveal limitations with respect to the assessment of mood. Some reports, despite omitting details of how mentation was assessed, still comment on the presence or absence of depression. Others suffer from one or more of the following shortcomings: a failure to assess premorbid risk factors for mood disorder; a reliance on one question to assess depression; the utilisation of self report mood rating scales of questionable validity; neglecting to distinguish depression as a symptom from depression as a syndrome (i.e. major depression as defined by the DMS-1V). CONCLUSIONS: Given the many methodological pitfalls inherent in all studies to date, it is premature to conclude that disease modifying drugs are associated with depression. Evidence suggests that treatment of depression, irrespective of a putative association with a disease modifying agent, is frequently effective. This applies to pharmacotherapy or psychotherapy, although the former may be preferred should depression arise during a course of treatment with a disease modifying agent. Multiple Sclerosis (2000) 6 343 - 348 PMID- 11064446 TI - Interferon-beta treatment for patients with multiple sclerosis: the patients' perceptions of the side-effects. AB - The aims of this study were to investigate (i) the self-reported frequency and intensity of systemic side-effects and their impact on the daily lives of patients suffering from Multiple Sclerosis (MS) and undergoing interferon-beta therapy and (ii) the self-reported frequency and perceptions of any local-tissue reactions. Forty patients aged 22 - 59 years (27 females) with relapsing/remitting MS were consecutively recruited for the study (17 on interferon-beta-1a and 23 on interferon-beta-1b). Two self-administered questionnaires were used before and after 1, 4, 8 and 16 weeks of therapy. The interferon therapy was found to be associated with flu-like symptoms. Most systemic side-effects were reported to be mild and to have little impact on the patients' daily lives. Asthenia and fatigue were more often rated as moderate or severe. The most frequently reported local-tissue side-effects were redness and local pain at the injection sites. A considerable inter-individual variation was found among patients regarding the perceptions of both the systemic and local side-effects. This suggests that it is of importance to identify early those patients who may need more support or other interventions to maintain a successful compliance. Multiple Sclerosis (2000) 6 349 - 354 PMID- 11064447 TI - MS-CANE: a computer-aided instrument for neurological evaluation of patients with multiple sclerosis: enhanced reliability of expanded disability status scale (EDSS) assessment. AB - Kurtzke's EDSS remains the most widely-used measure for clinical evaluation of MS patients. However, several studies have demonstrated the limited reliability of this tool. We introduce a computerized instrument, MS-CANE (Multiple Sclerosis Computer-Aided Neurological Examination), for clinical evaluation and follow up of patients with multiple sclerosis (MS) and to compare its reliability to that of conventional Expanded Disability Status Scale (EDSS) assessment. We developed a computerized interactive instrument, based on the following principles: structured gathering of neurological findings, reduction of compound notions to their basic components, use of precise definitions, priority setting and automated calculations of EDSS and functional systems scores. An expert panel examined the consistency of MS-CANE with Kurtzke's specifications. To determine the effect of MS-CANE on the reliability of EDSS assessment, 56 MS patients underwent paired conventional EDSS and MS-CANE-based evaluations. The inter observer agreement in both methods was determined and compared using the kappa statistic. The expert panel judged the tool to be compatible with the basic concepts of Kurtzke's EDSS. The use of MS-CANE increased the reliability of EDSS assessment: Kappa statistic was found to be 0.42 (i.e. moderate agreement) for conventional EDSS assessment versus 0.69 (i.e. substantial agreement) for MS-CANE (P=0.002). We conclude that the use of this tool may contribute towards a standardized and reliable assessment of EDSS. Within clinical trials, this could increase the power to detect effects, thus reducing trial duration and the cohort size required. Multiple Sclerosis (2000) 6 355 - 361 PMID- 11064448 TI - Long term azathioprine fails to prevent onset of multiple sclerosis: report of two cases. AB - Azathioprine is an immunosuppressive drug widely used in the treatment of chronic inflammatory diseases, including Multiple Sclerosis (MS). We report two patients who developed the first manifestations of clinically definite multiple sclerosis while on long term (3.5 and 10 years, respectively) treatment with azathioprine for Crohn's disease. Both patients developed the first MS symptoms during a quiescent phase of their inflammatory bowel disease. These cases show that long term azathioprine, while possibly maintaining inflammatory bowel disease under control, could not prevent the onset of MS. Multiple Sclerosis (2000) 6 362 - 363 PMID- 11064449 TI - Downregulation of telomerase reverse transcriptase mRNA expression by wild type p53 in human tumor cells. AB - The p53 tumor suppressor protein inhibits the formation of tumors through induction of cell cycle arrest and/or apoptosis. In the present study we demonstrated that p53 is also a powerful inhibitor of human telomerase reverse transcriptase (hTERT), a key component for telomerase. Activation of either exogenous temperature-sensitive (ts) p53 in BL41 Burkitt lymphoma cells or endogenous wild type (wt) p53 at a physiological level in MCF-7 breast carcinoma cells triggered a rapid downregulation of hTERT mRNA expression, independently of the induction of the p53 target gene p21. Co-transfection of an hTERT promoter construct with wt p53 but not mutant p53 in HeLa cells inhibited the hTERT promoter activity. Furthermore, the activation of the hTERT promoter in Drosophila Schneider SL2 cells was completely dependent on the ectopic expression of Sp1 and was abrogated by wt p53. Finally, wt p53 inhibited Sp1 binding to the hTERT proximal promoter by forming a p53-Sp1 complex. Since activation of telomerase, widely observed in human tumor cell lines and primary tumors, is a critical step in tumorigenesis, wt p53-triggered inhibition of hTERT/telomerase expression may reflect yet another mechanism of p53-mediated tumor suppression. Our findings provide new insights into both the biological function of p53 and the regulation of hTERT/telomerase expression. PMID- 11064450 TI - AP-1 complex is effector of Hox-induced cellular proliferation and transformation. AB - Hox gene products, initially characterized as master regulators of embryonic patterning, are also required for proper functioning of adult tissues. There is also a growing body of evidence that links Hox proteins to regulation of cellular proliferation/transformation. However, the underlying molecular mechanisms of Hox associated transformation and tissue growth have yet to be elucidated. Using a well established model system for studying changes in cellular proliferation induced by Hoxb4, we show that AP-1 activity is markedly increased in Hoxb4 transduced cells due to significant upregulation of Jun-B and Fra-1 protein levels. Furthermore, we also show that the specific changes in AP-1 protein expression are necessary for the proliferation effects induced by Hoxb4, and that these changes converge to increase levels of cyclin D1, a known integrator of proliferation signals. Our observations thus link Hox gene products with key elements of the cell cycle machinery. PMID- 11064451 TI - CL100/MKP-1 modulates JNK activation and apoptosis in response to cisplatin. AB - Treatment of cells with cisplatin induces a sustained activation of the stress activated protein kinase SAPK/JNK and the mitogen-activated protein kinase p38. Activation of JNK by cisplatin is necessary for the induction of apoptosis. Expression of the MAPK phosphatases CL100/MKP-1 and hVH-5 selectively prevents JNK/SAPK activation by cisplatin in a dose dependent fashion and results in protection against cisplatin-induced apoptosis. In contrast, expression of the ERK-specific phosphatase Pyst1 inhibits JNK/SAPK activity only when expressed at very high levels and does not confer protection against cisplatin. Furthermore, expression of a catalytically inactive mutant of CL100 in 293 cells decreases the IC50 for cisplatin and increases the toxicity of transplatin. This effect seems to be mediated by an increase in JNK activity since p38 activity is unaffected. These results suggest that dual-specificity MAPK phosphatases may be candidate drug targets in order to optimize cisplatin based therapeutic protocols. PMID- 11064452 TI - IL-15/IL-15Ralpha intracellular trafficking in human melanoma cells and signal transduction through the IL-15Ralpha. AB - There are two IL-15 isoforms and eight isoforms for the IL-15Ralpha chain whose biological role is poorly understood. Here, we have analysed the intracellular trafficking of IL-15 and IL-15Ralpha and tried to shed some light on their function(s). In IL-15/GFP CHO transfectants both IL-15 isoforms show nuclear localization. Two melanoma cell lines (MELP and MELREO) spontaneously expressing the IL-15 isoforms, display different intracellular trafficking of the IL-15/IL 15Ralpha complex. In MELP cells only IL-15Ralpha is detected inside the nucleus, whereas IL-15 and IL-15Ralpha assemble at the cell surface and are internalized. Moreover, the transducing molecule TRAF2 co-immunoprecipitates with IL-15Ralpha and may be deflected to TNFRI using anti-IL-15 blocking mAbs and TNF-alpha. By contrast, MELREO cells display IL-15Ralpha and IL-15 nuclear localization but only a partial co-localization of these molecules on the cell surface. In these cells, TRAF2 is strongly associated with IL-15Ralpha and cannot be deflected by any treatment. Since TRAF2 activates the transcription factor NF-kappaB, IL-15 through IL-15Ralpha, could have a role in the control of this pathway. Indeed, anti-IL-15 MaB inhibit the constitutive nuclear localization of NFkappaB and the phosphorylation of its inhibitor Ikappa-Balpha. Thus, IL-15Ralpha controls NF kappaB activation, however differences in the intracellular trafficking of the IL 15 and/or IL-15Ralpha suggest a different biological role for this complex in MELP versus MELREO cells. PMID- 11064453 TI - Chemotherapeutic drug, adriamycin, restores the function of p53 protein in hepatitis B virus X (HBx) protein-expressing liver cells. AB - Hepatitis B virus X (HBx) protein implicated in the development of liver cancer may inhibit the function of p53 tumor suppressor protein through cytoplasmic retention of p53 protein. Here, we attempt to investigate whether the functional inhibition of p53 protein by HBx protein is reversible. First, we provide the evidence for the association of endogenous p53 protein with HBx by co immunoprecipitation in stable Chang cells that express HBx protein in an inducible manner (ChangX-34). By immunofluorescence microscopy, the major location of p53 protein of ChangX-34 cells was confirmed at the nuclear periphery as well as in the cytoplasm where HBx protein is mainly expressed. Surprisingly, anticancer drug, adriamycin induces the nuclear translocation of p53 protein sequestered in the cytoplasm. This change is accompanied by the restoration of p53 activity, which results in increased transcriptional activity at the p53 responsive DNA elements as well as increase of p21WAF1 mRNA expression. Further, we observed the induction of cell death and G1 arrest in these cells upon adriamycin treatment regardless of HBx expression. Together, we demonstrate that functional inhibition of p53 protein through its cytoplasmic retention by HBx protein is reversible. These results may be extended into other tumors of which p53 activity is modulated by viral oncoproteins. PMID- 11064454 TI - Stimulation of tumor-associated fatty acid synthase expression by growth factor activation of the sterol regulatory element-binding protein pathway. AB - Increased expression of fatty acid synthase (FAS) is observed in a clinically aggressive subset of various common cancers and interference with FAS offers promising opportunities for selective chemotherapeutic intervention. The mechanisms by which FAS expression is (up)-regulated in these tumors remain, however, largely unknown. Recently we demonstrated that in LNCaP prostate cancer cells FAS expression is markedly elevated by androgens via an indirect pathway involving sterol regulatory element-binding proteins (SREBPs). Here, we also show that growth factors such as EGF are able to stimulate FAS mRNA, protein and activity. Several observations also indicate that the effects of EGF on FAS expression are ultimately mediated by SREBPs. EGF stimulates SREBP-1c mRNA expression and induces an increase in mature nuclear SREBP-1. Moreover, in transient transfection studies EGF stimulates the transcriptional activity of a 178 bp FAS promoter fragment harboring a complex SREBP-binding site. Deletion or mutation of this binding site abolishes these effects and ectopic expression of dominant negative SREBP-1 inhibits FAS expression and induction in intact LNCaP cells. Given the frequent dysregulation of growth factor signaling in cancer and the key role of SREBP-1 in lipid homeostasis, growth factor-induced activation of the SREBP pathway is proposed as one of the mechanisms responsible for up regulation of lipogenic gene expression in a subset of cancer cells. PMID- 11064455 TI - Sp1 and Sp3 activate p21 (WAF1/CIP1) gene transcription in the Caco-2 colon adenocarcinoma cell line. AB - The CDK inhibitor p21WAF1/CIP1 is a negative regulator of the cell cycle, and its expression is induced during terminal differentiation in vitro and in vivo. Expression of p21 is controlled at the transcriptional level by both p53 dependent and -independent mechanisms. Our previous studies established that p21 is expressed in the Caco-2 adenocarcinoma cell line, and its expression is induced by a p53-independent mechanism during differentiation of these cells. Here we have found that transcription of p21 in Caco-2 cells is controlled primarily by the transcription factors Sp1 and Sp3 through two Sp1 binding sites, Sp1-1 and Sp1-2, located between -119 and -114 bp and between -109 and -104 bp of the p21 promoter, respectively. Sp1 and Sp3 binding to the p21 promoter increased during Caco-2 cell differentiation, while the absolute level of Sp1 did not change and the absolute level of Sp3 increased approximately twofold. Transfection experiments in the SL2 Drosophila cell line that lacks endogenous Sp3 activity demonstrated that Sp1 transactivates the p21 promoter primarily through the Sp1-2 site, while Sp3 acts through the Sp1-1 site. In these cells Sp3 is a stronger transactivator of the p21 promoter than Sp1. Our data suggest that induction of p21 transcription during Caco-2 differentiation is modulated by Sp1/Sp3 interactions with the p21 promoter. PMID- 11064456 TI - Differential activity of conditional MYC and its variant MYC-S in human mortal fibroblasts. AB - We have explored the effects of the conditional MYC-estrogen receptor fusion protein, MYC-ERTM, in human mortal fibroblasts, WI38, on cell-cycle entry, apoptosis and gene expression. The results indicate that activation of MYC-ERTM in WI38 cells is sufficient to cause S phase entry of quiescent cells, which is preceded by phosphorylation of Rb and activation of the Cdk2-associated kinase. We also analysed the MYC protein variant, MYC-S, which lacks part of the transcriptional activation domain but includes the conserved MYC box II and 26 amino acids N-terminal to it. MYC-S was previously shown to promote proliferation and apoptosis of immortalized rodent cell lines. The results indicate that MYC-S has undetectable activity as an inducer of S phase or apoptosis of quiescent WI38 cells. However, Myc-S stimulates proliferation of WI38 cells in the presence of 10% fetal calf serum. Surprisingly, we found that MYC-S, previously considered solely a repressor of specific reporter genes, is instead a weak transactivator of endogenous target genes both in mortal and immortalized cells. In addition, MYC-S exhibit a weak repressor activity upon an endogenous target gene only in immortalized cells. MYC-S transcriptional properties suggest that MYC box II and the adjacent N-terminal amino acids, while not sufficient for full repression function, participate in transactivation of endogenous target genes. PMID- 11064457 TI - Activation of IKKalpha and IKKbeta through their fusion with HTLV-I tax protein. AB - Human T-cell leukemia virus type I (HTLV-I) Tax protein persistently stimulates the activity of IkappaB kinase (IKK), resulting in constitutive activation of the transcription factor NF-kappaB. Tax activation of IKK requires physical interaction of this viral protein with the IKK regulatory subunit, IKKgamma. The Tax/IKKgamma interaction allows Tax to engage the IKK catalytic subunits, IKKalpha and IKKbeta, although it remains unclear whether this linker function of IKKgamma is sufficient for supporting the Tax-specific IKK activation. To address this question, we have examined the sequences of IKKgamma required for modulating the Tax/IKK signaling. We demonstrate that when fused to Tax, a small N-terminal fragment of IKKgamma, containing its minimal IKKalpha/beta-binding domain, is sufficient for bringing Tax to and activating the IKK catalytic subunits. Disruption of the IKKalpha/beta-binding activity of this domain abolishes its function in modulating the Tax/IKK signaling. We further demonstrate that direct fusion of Tax to IKKalpha and IKKbeta leads to activation of these kinases. These findings suggest that the IKKgamma-directed Tax/IKK association serves as a molecular trigger for IKK activation. PMID- 11064458 TI - Human cortactin as putative cancer antigen. AB - Humoral immune response against overexpressed oncogenic or tumor supressor proteins has been demonstrated for many types of cancer. In this study we report on the detection of the autologous antibody response to putative oncogene, human cortactin using serological analysis of breast carcinoma expression library. Cortactin maps to chromosome 11q13, the region amplified in about 15% of primary breast carcinomas and 30% of head and neck squamous cell carcinomas. Cortactin overexpression due to such amplification might affect adhesive properties of human cancer cells and is associated with poor disease prognosis. Accordingly, we detected overexpression of cortactin transcript in autologous tumor and amplification/overexpression of cortactin in tumors of breast cancer patients serologically positive for this marker. We demonstrate that 15% of breast cancer patients elicit humoral immune response against human cortactin. PMID- 11064459 TI - The Ron oncogenic activity induced by the MEN2B-like substitution overcomes the requirement for the multifunctional docking site. AB - Oncogenic activation of the Ron tyrosine kinase (Macrophage Stimulating Protein receptor) relies on substitutions of two highly conserved residues in the catalytic domain (D1232V and M1254T), which result in ligand-independent activation of the receptor, in vivo tumorigenesis and metastasis. We show here that the Y/F conversion of the Y1317 residue in the kinase domain impairs tumorigenic and metastatic properties of Ron activated by the MEN2B-like mutation (RonM1254T), but not by other two oncogenic substitutions. Furthermore, RonM1254T lacking the multifunctional docking site retains transforming and metastatic activity. These data reveal that the transforming activity of RonM1254T mutant is dependent on Y1317 phosphorylation, suggesting a shift in intramolecular substrate specificity. Consistently, a shift of RonM1254T kinase substrate specificity was observed by in vitro peptide phosphorylation assays and in vivo receptor auto-phosphorylation. The Y1317 phosphorylation elicits by itself activation of PI-3K/Akt and MAPK signalling pathways. Our data indicate that the accomplishment of the full oncogenic phenotype of RonM1254T requires the phosphorylation both of the canonical C-terminal docking site and of the unique Y1317 residue in the tyrosine kinase domain. PMID- 11064460 TI - The Italian registry of myelofibrosis is one year old. PMID- 11064461 TI - Advances in iron chelating therapy. PMID- 11064462 TI - Antitumor vaccination. PMID- 11064463 TI - Relevance of bone marrow features in the differential diagnosis between essential thrombocythemia and early stage idiopathic myelofibrosis. AB - BACKGROUND AND OBJECTIVES: Diagnosis of essential thrombocythemia (ET) remains a challenging problem and has been predominantly established by exclusion of other thrombocythemic disorders. In this context the updated diagnostic criteria of the Polycythemia Vera Study Group (PVSG) are generally accepted, although histopathologic features of the bone marrow were only marginally considered. DESIGN AND METHODS: A retrospective evaluation was performed of 168 patients presenting with ET in accordance with the criteria of the PVSG. Analysis was focused on the discriminating impact of bone marrow morphology. RESULTS: Histopathology revealed that our cohort of patients could be divided into three distinct groups (true ET, questionable ET and false ET). These groups were characterized by certain diagnostic constellations of clinical data on admission. True ET was found in 53 patients presenting with no or a borderline splenomegaly and no relevant anemia or leuko-erythroblastic blood picture. The other patients showed clinical signs and symptoms which were more compatible with initial prefibrotic (52 patients) or early (68 patients) idiopathic-primary myelofibrosis (IMF) with severe thrombocythemia. In true ET no significant hypercellularity of the bone marrow including myeloid precursors or an increase in reticulin fibers was detectable. Most prominent were changes of megakaryopoiesis which revealed large to giant-sized cells lacking a definite maturation defect. Their appearance in true ET contrasted with the clusters of abnormally differentiated, often bizarre elements of this lineage in patients with initial and early IMF (questionable or false ET). Calculation of survival disclosed a relevant disparity with a non-significant loss in life expectancy of 10.9% in true ET compared to 29.6% in questionable and 51.3% in false ET. Follow-up studies and repeated bone marrow biopsies revealed no transition into myelofibrosis in true ET, whereas this did occur in 22 of 27 patients with questionable and false ET. In the latter cohort bone marrow changes were accompanied by increasing anemia, splenomegaly, tear-drop poikilocytosis and reduction of the platelet count consistent with IMF. INTERPRETATION AND CONCLUSIONS: A detailed evaluation of bone marrow features, in particular megakaryopoiesis is recommended to establish positive criteria for the diagnosis of ET and thus to accomplish a significant improvement of the PVSG postulates. In this context ongoing clinical trials on ET must regard pretreatment bone marrow biopsies as a major clue to diagnosis. PMID- 11064464 TI - Long-term follow-up after fludarabine treatment in pretreated patients with chronic lymphocytic leukemia. AB - BACKGROUND AND OBJECTIVES: A study update to assess long-term survival following fludarabine salvage treatment in previously treated patients with chronic lymphocytic lymphoma (CLL). DESIGN AND METHODS: From September 1992 to December 1995, 74 patients with advanced, relapsing B-cell CLL were enrolled in the study. Fludarabine was given for 5 consecutive days at the dose of 25 mg/m2/day in a 30 min infusion. Treatment was repeated every 28 days for a maximum of 6 courses.E RESULTS. Nineteen (26%) patients achieved a complete response (CR) and 20 (27%) patients had a partial response (PR), giving an overall response rate of 53%. The median overall survival was 68 months, and there was a strong negative correlation with the number of previous treatments. The median time to progression was 18 months for patients who achieved a CR and 12 months for those with a PR. INTERPRETATION AND CONCLUSIONS: The results obtained with fludarabine alone in this subset of CLL patients indicate the existence of a conspicuous disease-free survival period. This time window could be used to consolidate the initial response with either biological approaches or high-dose therapeutic strategies such as autologous bone marrow transplantation, with the aim of eventual eradication of the disease. PMID- 11064465 TI - Quantitative expression of CD23 and its ligand CD21 in chronic lymphocytic leukemia. AB - BACKGROUND AND OBJECTIVES: Cells from the great majority of patients with chronic lymphocytic leukemia (CLL) express CD23. A recent histologic study has shown that CD23 is expressed more strongly in the proliferating centers of the lymph nodes, where the large prolymphocytoid cells are located. The aim of our study was to quantify the expression of CD23 and CD21 in small and prolymphocytoid cells from patients with CLL and B-cell lymphomas, and correlate this expression with clinical parameters. DESIGN AND METHODS: Using quantitative flow cyto-metry we analyzed the antigen density of CD23 and CD21 in: 1) 101 cases of chronic lymphocytic leukemia, 84 typical, 14 with increased prolymphocytes (CLL/PL) and 3 atypical, 2) 15 cases of CD23 positive B-cell lymphoma with circulating lymphoma cells and 3) 8 normal subjects. The results were correlated with morphology and clinical staging. RESULTS: Cells from CLL and CLL/PL have a significantly higher number of CD23 molecules than normal and lymphoma B-cells (p<0.001 and p<0.001, respectively). Differences were not significant for CD21. CLL and CLL/PL cases had similar values of CD23 and CD21 molecules, but analysis at a single level showed that prolymphocytes in typical CLL and CLL/PL expressed significantly higher CD23 (p=0.001, p=0.006) and CD21 (p=0.001, p=0.001) than small lymphocytes. There was no correlation between CD23 or CD21 antigen density and clinical stages although there was a trend for a brighter CD23 in stage C patients. INTERPRETATION AND CONCLUSIONS: Since interaction between CD23 and CD21 is important for B-cell activation, proliferation and tumor formation, findings that both molecules are upregulated in prolymphocytes suggest that this is the proliferating cell component in CLL and underline the association between progression and increased prolymphocytes in typical CLL and CLL/PL. PMID- 11064466 TI - Correlation between cytogenetic abnormalities and disease characteristics in multiple myeloma: monosomy of chromosome 13 and structural abnormalities of 11q are associated with a high percentage of S-phase plasma cells. AB - BACKGROUND AND OBJECTIVES: Cytogenetic studies in multiple myleoma (MM) are limited by the difficulties in obtaining metaphases that can be investigated and few studies have analyzed the relationship between cytogenetics and clinical disease characteristics. The aim of our study was to analyze the recurrent cytogenetic changes in MM and to correlate them with clinical and biological characteristics including the percentage of S-phase plasma cells (PCs). DESIGN AND METHODS: Chromosomal abnormalities were analyzed in 86 patients with MM. In all patients, two types of cultures (5 d culture with interleukin-4 and unstimulated 72 h culture) were used for cytogenetic analysis. DNA content analysis (ploidy and cell cycle analysis) together with the most relevant clinical and biological disease features were studied. RESULTS: Cytogenetic analysis was successful in 72 of the 86 patients (84%). Forty-seven patients (65%) had an abnormal karyotype. The most frequent trisomies involved chromosomes 3, 5, 9, 11, 15, 19, 22, 1, 7, 17, 18, and 21, and monosomies affected chromosomes 13 and 8, while structural changes involved chromosomes 1, 11, 14q32, 4p16 and 16q22-23. Patients with abnormal karyotype displayed a poor performance status, advanced stage, anemia and a high percentage of bone marrow plasma cells. In addition, MM patients with -13/13q- and 11q abnormalities showed a significantly higher proportion of S-phase PCs (p=0.02). INTERPRETATION AND CONCLUSIONS: In summary, our study shows a relationship between unfavorable cytogenetics (-13/13q-/11q abnormalities) and a high percentage of S-phase PCs, a well-known adverse prognostic factor. PMID- 11064467 TI - Need for an accurate molecular diagnosis to assess the donor origin of leukemia relapse after allogeneic stem cell transplantation. AB - BACKGROUND AND OBJECTIVES: Leukemia relapse occurring in donor cells after allogeneic hematopoietic stem cell transplantation has been reported in rare cases. Cytogenetic analysis and molecular probing of variable number of tandem repeats (VNTRs) have been used to confirm this unusual event in the few cases so far reported in the literature. The aim of this study was to demonstrate that extensive molecular characterization of leukemic cells at diagnosis and relapse may be necessary to avoid many technical pitfalls possibly leading to an erroneous diagnosis of leukemia relapse in donor cells after allogeneic transplantation. DESIGN AND METHODS: We report the case of a 49- year old man who received an allogeneic transplantation from his HLA-identical sister because of BCR-ABL+ acute lymphoblastic leukemia (ALL). After having achieved complete hematologic and molecular remission, two years later an overt leukemia relapse occurred with cytogenetic findings suggesting a leukemia relapse in donor cells. The donor or patient origin of leukemic cells at relapse was further investigated by fluorescence in situ hybridization (FISH) karyotyping, reverse transcription (RT) polymerase chain reaction (PCR) analysis of BCR-ABL chimeric transcripts, PCR amplification of several VNTRs and the Y chromosome-specific DYS14 sequence and finally by amplification, cloning and sequencing of the CDRIII region of the immunoglobulin heavy chain (IgH) gene. RESULTS: At the time of relapse, conventional and FISH karyotyping revealed the presence of a Phl+ chromosome and a female karyotype in all the 25 metaphases analyzed and PCR amplification of the Y chromosome-specific DYS14 sequence was negative. Moreover, the molecular evaluation of hematopoietic chimerism performed by the NZ-22 VNTR allowed us to demonstrate that at the time of relapse, a consistent proportion of hematopoietic cells was of donor origin. However, the molecular cloning and sequencing of the CDRIII region of the immunoglobuin heavy chain (IgH) gene rearrangement in leukemic blasts at diagnosis and relapse demonstrated their identity thus formally proving the patient origin of both leukemic clones. INTERPRETATION AND CONCLUSIONS: While the simplest interpretation of the apparent female karyotype at relapse is the consequence of a loss of the Y chromosome which in leukemic blasts took place along with duplication of an X-chromosome, this case strongly emphasizes the need for accurate and extensive molecular characterization to prove the donor origin of a leukemia relapse after allogeneic transplantation. PMID- 11064468 TI - Vancomycin-resistant Enterococcus faecium infection in three children given allogeneic hematopoietic stem cell transplantation: clinical and microbiologic features. AB - BACKGROUND AND OBJECTIVES: The emergence of vancomycin-resistant enterococci (VRE) as nosocomial pathogens is a major problem in the US; in Europe, VRE nosocomial infections are uncommon and only rarely have been reported in Pediatric or Neonatal Units. The aim of this study is to report on the clinical and microbiological features of VRE infections in 3 children given hematopoietic stem cell transplantation (HSCT). PATIENTS AND METHODS: Five episodes of vancomycin-resistant Enterococcus faecium (VREF) infection were diagnosed in 3 children given an allogeneic HSCT. Molecular methods, such as random amplification of polymorphic DNA (RAPD) fingerprinting and automated ribotyping, were used in order to define the circulation of strains. RESULTS: All the isolates were resistant to all commercially available agents and showed the VanA genotypic profile. All children were successfully treated with the combination of quinupristin/dalfopristin (QD) plus teicoplanin (TEC), although treatment was not sufficient to eradicate the micro-organism promptly from the gastrointestinal tract. All our children are still alive. After the first isolation of VRE, a surveillance protocol was started and we documented that the rate of colonization in children and their mothers was less than 1.5%. The RAPD method demonstrated the possible nosocomial transmission of one strain. INTERPRETATION AND CONCLUSIONS: Our experience demonstrates that VRE infection is a life-threatening complication in children given HSCT. Prompt diagnosis of this infection and its treatment with the combination of QD and TEC can successfully manage this severe infection in profoundly immunocompromised patients. PMID- 11064469 TI - Allogeneic peripheral blood stem cell transplantation with CD34+-cell selection and delayed T-cell add-back in adults. Results of a single center pilot study. AB - BACKGROUND AND OBJECTIVES: Allogeneic peripheral blood stem cell transplantation with CD34+ cell-selection (CD34+-PBSCT) allows rapid hematologic engraftment with a reduction in graft-versus-host disease (GVHD), although concerns exist regarding the increased risk of tumor relapse associated with T-cell depletion of the graft. Delayed T-cell add-back (TCAB) after such transplants may restore the graft-versus-tumor effect while achieving a reduced early transplant-related mortality due to less GVHD in a group of patients at high risk of early death (i.e., age >= 45 years). DESIGN AND METHODS: Ten patients 45 years of age or older with hematologic malignancies received a CD34+-PBSCT and cyclosporin A (CyA) to prevent acute GVHD, followed by a planned delayed donor TCAB of 107 T cells/kg to restore the graft-versus-tumor effect. The infused graft included a median of 6.3x106 CD34+ cells/kg and 4.4x104 CD3+ cells/kg. RESULTS: Engraftment was prompt in all cases. Four patients developed acute GVHD after the CD34+-PBSCT and/or chronic GVHD after CyA withdrawal and did not proceed to TCAB, and two patients died early before the planned TCAB. Four patients proceeded to TCAB at a median of day +104 after CD34+-PBSCT (+92 to +150). Two of these patients developed acute GVHD grades I-II (IBMTR Index B) after TCAB and all four developed chronic GVHD, which was extensive in two. With a median follow-up of 611 days (range 499-847) after transplant in the seven survivors, there have been no disease progressions, and all patients show a pattern of complete donor chimerism in bone marrow and peripheral blood. INTERPRETATIONS AND CONCLUSIONS: The results of our pilot study suggest that this protocol produces an acceptable transplant-related morbidity and mortality in patients 45 years and older. However, there may be benefit in infusing CD34+-selected PBSCT with even lower T cell contents and further delaying the TCAB. PMID- 11064470 TI - Cytogenetic characterization of acute myeloid leukemia in Shwachman's syndrome. A case report. AB - We report on a case of acute myeloid leukemia in a 17-year old boy affected by Shwachman Diamond syndrome (SDS). Conventional cytogenetics at diagnosis revealed an abnormal clone with complex karyotypic changes including typical myeloid aberrations, such as monosomy 5, tetrasomy of chromosome 8, trisomy 9, and deletion of the short arm of chromosome 12. The boy was treated with conventional chemotherapy and reached complete remission of leukemia, confirmed by cytogenetics and fluorescence in situ hybridization. Nevertheless he failed to regenerate normal marrow cellularity and blood cell count. Cytogenetic information on hematologic malignancies in SDS patients are discussed. PMID- 11064471 TI - Isolated spleen agenesis: a rare cause of thrombocytosis mimicking essential thrombocythemia. AB - Thrombocytosis is a common feature of myeloproliferative disorders but may also result from various conditions including chronic iron deficiency, hemorrhage, chronic inflammation and splenectomy. We report two cases of secondary thrombocytosis caused by isolated and congenital asplenia, mimicking essential thrombocythemia. These two adult cases of spleen agenesis were unexpected. We conclude that in thrombocytosis without clinical evidence of splenomegaly, attentive screening of blood in search of Howell-Jolly bodies and abdominal ultrasonography should always be performed not only to detect mild spleen enlargement but also to make sure of the presence of this organ. PMID- 11064472 TI - The irreplaceable image: Scintigraphic imaging with (99m)Tc-HMPAO-labeled leukocytes in neutropenia due to increased neutrophil sequestration. PMID- 11064473 TI - Thalassemic trait caused by IVS II-1 (GAEA) mutation detected in a Spanish family. PMID- 11064474 TI - Azoospermia in a patient with sickle cell disease treated with hydroxyurea. PMID- 11064475 TI - Further evidence on the underestimation of the prevalence of TT viral DNA in blood donors. PMID- 11064476 TI - Karyotype refinement in five patients with acute myeloid leukemia using spectral karyotyping. PMID- 11064477 TI - Acute leukemia in Jehovah's Witnesses: a challenge for hematologists. PMID- 11064478 TI - Telomerase: obviously activated in the accelerated phase of chronic myeloid leukemia. PMID- 11064479 TI - Fludarabine therapy in chronic lymphocytic leukemia-associated severe nephrotic syndrome. PMID- 11064480 TI - 8q24 translocations in blastic transformation of mantle cell lymphoma. PMID- 11064481 TI - Comparative analysis of immunophenotypic methods for the assessment of minimal residual disease in hairy cell leukemia. PMID- 11064482 TI - Oral glutamine supplements in autologous hematopoietic transplant: impact on gastrointestinal toxicity and plasma protein levels. PMID- 11064483 TI - A boy with venous thrombosis, homozygous for factor V Leiden, prothrombin G20210A and MTHFR C667t mutations, but belonging to an asymptomatic family. PMID- 11064484 TI - An overview of heroin trends in New York City: past, present and future. AB - Heroin abuse has been a long-standing problem in the United States, especially in New York City, which is a major heroin trafficking center and home to the largest population of heroin addicts in the country. As a consequence, New York City is also the major center for methadone treatment. Over time, however, the heroin problem seems to have changed in terms of demographic characteristics of abusers, as well as heroin purity, its associated health consequences, modes of use, and other drug patterns. Past and present heroin-involved events for New York City are examined, with some projection about the future of the heroin problem. To identify these changes, a variety of indirect indicators of heroin abuse are analyzed. These indirect indicators include admissions to New York City heroin treatment programs, as well as heroin-involved deaths, arrests and emergency room episodes, and heroin purity levels. Additionally, a recent study of heroin abusers in New York City yielded patterns of heroin use and estimates of heroin prevalence. The salient results indicate that, over time, Hispanics have become the predominant user group, the population of abusers is an aging cohort, heroin purity levels have risen dramatically, and intranasal use has become more prominent than injecting. The findings have important consequences for prevention, treatment, and programs that target special populations. PMID- 11064485 TI - Methadone maintenance treatment (MMT): a review of historical and clinical issues. AB - Methadone maintenance has been evaluated since its development in 1964 as a medical response to the post-World War II heroin epidemic in New York City. The findings of major early studies have been consistent. Methadone maintenance reduces and/or eliminates the use of heroin, reduces the death rates and criminality associated with heroin use, and allows patients to improve their health and social productivity. In addition, enrollment in methadone maintenance has the potential toreduce the transmission of infectious diseases associated with heroin injection, such as hepatitis and HIV. The principal effects of methadone maintenance are to relieve narcotic craving, suppress the abstinence syndrome, and block the euphoric effects associated with heroin. A majority of patients require 80-120 mg/d of methadone, or more, to achieve these effects and require treatment for an indefinite period of time, since methadone maintenance is a corrective but not a curative treatment for heroin addiction. Lower doses may not be as effective or provide the blockade effect. Methadone maintenance has been found to be medically safe and nonsedating. It is also indicated for pregnant women addicted to heroin. Reviews issued by the Institute of Medicine and the National Institutes of Health have defined narcotic addiction as a chronic medical disorder and have claimed that methadone maintenance coupled with social services is the most effective treatment for this condition. These agencies recommend reducing governmental regulation to facilitate patients access to treatment. In addition, they recommend that the number of programs be expanded, and that new models of treatment be implemented,if the nationwide problem of addiction is to be brought under control. The National Institutes of Health also recommend that methadone maintenance be available to persons under legal supervision, such as probationers, parolees and the incarcerated. However, stigma and bias directed at the programs and the patients have hindered expansion and the effective delivery of services. Professional community leadership is necessary to educate the general public if these impediments are to be overcome. PMID- 11064486 TI - The cost-effectiveness of methadone maintenance. AB - BACKGROUND: Although methadone maintenance is effective in reducing injection drug use, needle sharing, and the overall mortality associated with opiate abuse, many health plans offer little or no access to methadone, and many methadone providers do not comply with treatment guidelines regarding dose, duration of treatment, or provision of ancillary services. Moral and political judgments have helped shape the U.S. treatment system. Evaluations of methadone cost effectiveness may play a role in changing public policy. METHOD: Cost effectiveness analysis is used to compare a change, or changes, in treatment to that of current standard care. The cost of treatment and its effect on outcomes are used to find the incremental cost-effectiveness ratio, and determine whether the change(s) should be adopted. The literature on methadone maintenance is reviewed from an economic perspective, focusing on five policy questions: (1) whether methadone should be a health care benefit; (2) what level of ancillary services is optimal; (3) what methadone dose is appropriate; (4) what length of treatment is appropriate; and (5) whether contingency contracts should be employed. RESULTS: Expanded access to methadone maintenance has an incremental cost-effectiveness ratio of less than $11,000 per Quality-Adjusted Life Year. This is more cost-effective than many widely used medical therapies, a finding that strongly supports the inclusion of methadone in the formulary of health care plans.Ancillary services have been shown to be an effective part of methadone maintenance therapy, especially during the beginning of a treatment episode, but there is not enough information available to tell whether the optimal amount of services is being used. There is extensive evidence that many treatment programs dispense inadequate doses of methadone. The cost of additional drugs is very small compared to the benefits of an adequate dose. Many methadone programs limit treatment to 6 months or less, but such short episodes are not likely to be cost effective. The medical model of methadone maintenance may increase the cost effectiveness of the treatment for long-term patients. Programs that reward patients for negative urinalysis have proven effective at reducing illicit drug use, but their cost-effectiveness will need to be demonstrated before they are widely adopted. CONCLUSIONS: Cost-effectiveness researchers need to measure substance abuse outcomes in terms of Quality-Adjusted Life Years, as this will make their findings more relevant to the development of treatment policy. It will allow different substance abuse treatments to be compared to each other and to medical care interventions. PMID- 11064487 TI - Neurobiology of addictive behaviors and its relationship to methadone maintenance. AB - Scientific information about the neurobiology of addictive behaviors provides an increasingly important rationale to support opioid agonist pharmacotherapy, primarily methadone maintenance treatment, for long-term heroin addiction. In late 1963 and 1964, the first research was performed at The Rockefeller Institute for Medical Research by Dole, Nyswander, and Kreek in an attempt to develop a new pharmacotherapy for opiate addiction. The hypothesis underlying that research was that heroin addiction was a disease. However, the evidence for heroin addiction being a disease was based primarily on clinical anecdotes and the natural history of opiate addiction. Until then chronic addiction was managed primarily using abstinence-based, medication-free behavioral approaches. Such approaches were uniformly successful in only a small percent of long-term heroin addicts. Subsequent research, both clinical research as well as laboratory-based research, using a variety of appropriate animal models as well as in vitro techniques, has shown that drugs of abuse in general, and specifically the short-acting opiates, such as heroin, may profoundly alter molecular and neurochemical indices, and thus physiologic functions. Also, research has shown that after chronic exposure to a short-acting opiate,these alterations may be persistent, or even permanent, and may contribute directly to the perpetuation of self-administration of opiates, and even the return to opiate use after achieving a drug-free and medication-free state. There is ample evidence now that disruption of several components of the endogenous opioid system, ranging from changes in gene expression to changes in behavior, may occur during cycles of short-acting opiate abuse. Also, there are very convincing studies that suggest that stress responsivity is profoundly altered by chronic abuse of short-acting opiates including: documentation of atypical hypo-responsivity to stressors during cycles of heroin addiction; evidence of sustained hyper-responsivity to stressors in the medication-free, illicit-opiate-free state; and in contrast, normalization of stress responsivity, as reflected by the hypothalamic-pituitary-adrenal axis function in long-term, methadone-maintained patients. Thus, both laboratory and clinical research studies provide firm documentation that the disruption of physiologic, as well as behavioral, functions occurs during chronic administration of short-acting opiates. Also, there is research evidence of an epidemiologic, and more recently of a molecular genetics type, that a genetic vulnerability to develop addictions in general, and opiate addiction specifically, may exist, and that early environmental factors may alter physiology to enhance vulnerability to develop opiate addiction when self exposed. PMID- 11064488 TI - Cerebral metabolism in opiate-dependent subjects: effects of methadone maintenance. AB - BACKGROUND: The long-term effects of opiate use on human brain are not known. The goal of this preliminary study was to determine whether human subjects with histories of opiate dependence have persistent differences in brain function as compared with individuals without substance use disorders, and whether methadone maintenance reverses or ameliorates the potential abnormality. METHOD: Positron emission tomographic (PET) [18F]fluorodeoxyglucose (FDG) method was used to compare the regional cerebral metabolic rate for glucose(rCMRglc) in three groups: four opiate-dependent subjects currently receiving methadone maintenance therapy (MM), four opiate-dependent subjects not receiving methadone maintenance therapy (MW), and a comparison group of five subjects without substance use disorders. RESULTS: A significant difference in rCMRglc in the anterior cingulate gyrus was found between the MW and Control groups (Mann-Whitney U=2.0, p=0.05). Generally speaking, rCMRglc's in MM subjects were intermediate between those of MW and Control groups, although the difference did not reach statistical significance. CONCLUSIONS: The results of this study suggest that neurobiological abnormalities can persist in the brain of a chronic opiate user several years after detoxification from methadone. Future research is needed to replicate these results and to determine whether the observed rCMRglc differences are related to opiate use or to neurochemical abnormalities that play a role in developing addictive behavior. PMID- 11064489 TI - Methadone medical maintenance (MMM): treating chronic opioid dependence in private medical practice--a summary report (1983-1998). AB - BACKGROUND: Methadone Medical Maintenance (MMM) was implemented in 1983 to enable socially rehabilitated methadone patients to be treated in the offices of private physicians rather than in the traditional clinic system. Over a period of 15 years, 158 methadone patients who fulfilled specific criteria within the clinic system entered this program in New York City. Participating patients reported to their physician once a month and received a one-month supply of methadone tablets rather than a one-day liquid dose in a bottle. METHOD: Of the 158 patients who entered this program, 132 (83.5%) were compliant with the regulations and proved to be treatable within the hospital-based private practices of internists participating in the program. Compliant MMM patients found it easier to improve their employment status and business situations, finish their educations, and normalize their lives in MMM as opposed to the traditional clinic system because they had simplified reporting schedules and fewer clinical restrictions. Twelve (8%) compliant patients were able to successfully withdraw from methadone after an average of 17.7 years of treatment in both the traditional clinics and MMM. Twenty compliant patients (13%) died from a variety of causes, 40% of which were related to cigarette smoking. None of the deaths were attributable to long-term methadone treatment. Other causes of death included hepatitis C, AIDS, cancer, homicide, complications of morbid obesity and meningitis. RESULTS: The 26 noncompliant patients (16.5%) were referred back to their clinics for continued treatment or were discharged for failure to report as directed. A major cause of failure in MMM was abuse of crack/cocaine. CONCLUSIONS: Stigma concerning enrollment in methadone treatment was a major social issue that patients faced. Many refused to inform employers, members of their families, friends, and other physicians who treated them for a various of conditions that they were methadone patients. The methadone medical maintenance physician, therefore, functions as a medical ombudsman for the patient, educating other physicians who treat the patient about methadone maintenance and its applicability to the patient. Our results can serve as a model for the expansion of office-based MMM treatment. PMID- 11064490 TI - Effects of LAAM and methadone utilization in an opiate agonist treatment program. AB - The development and approval of levo-alpha-acetylmethadol (LAAM) as a pharmacotherapeutic agent in opioid agonist therapy provided an alternative to methadone. Clinicians recognized the potential benefits that LAAM, a synthetic mu agonist with pharmacological properties which differ from those of methadone,could have in the treatment management of addicts in opioid agonist therapy. We report our experience utilizing LAAM from 1995 to 1999 at the Hines VA opioid agonist therapy clinic. The addition of LAAM to the clinic's treatment armamentarium has resulted in management options that have improved the areas of patient recruitment, patient retention, patient traffic, take-home medication, detoxification, and treatment outcomes. PMID- 11064491 TI - When "enough" is not enough: new perspectives on optimal methadone maintenance dose. AB - Some methadone maintenance treatment (MMT) programs prescribe inadequate daily methadone doses. Patients complain of withdrawal symptoms and continue illicit opioid use, yet practitioners are reluctant to increase doses above certain arbitrary thresholds. Serum methadone levels (SMLs) may guide practitioners dosing decisions, especially for those patients who have low SMLs despite higher methadone doses. Such variation is due in part to the complexities of methadone metabolism. The medication itself is a racemic (50:50) mixture of 2 enantiomers: an active "R" form and an essentially inactive "S" form. Methadone is metabolized primarily in the liver, by up to five cytochrome P450 isoforms, and individual differences in enzyme activity help explain wide ranges of active R-enantiomer concentrations in patients given identical doses of racemic methadone. Most clinical research studies have used methadone doses of less than 100 mg/day [d] and have not reported corresponding SMLs. New research suggests that doses ranging from 120 mg/d to more than 700 mg/d, with correspondingly higher SMLs, may be optimal for many patients. Each patient presents a unique clinical challenge, and there is no way of prescribing a single best methadone dose to achieve a specific blood level as a "gold standard" for all patients. Clinical signs and patient-reported symptoms of abstinence syndrome, and continuing illicit opioid use, are effective indicators of dose inadequacy. There does not appear to be a maximum daily dose limit when determining what is adequately "enough" methadone in MMT. PMID- 11064492 TI - Treatment of pain in methadone-maintained patients. AB - Patients with opioid dependency experience trauma, acute medical illness and chronic diseases, and may have to undergo surgery to the same extent as other individuals. They need to be treated for relief of symptoms, including pain. Undertreatment or inadequate treatment of pain for these individuals is a particular problem because of opioid dependency and/or methadone maintenance treatment. The guiding principles governing treatment of these patients are to maintain the methadone treatment and to use short-acting narcotics administered at higher doses, and to do so as often as necessary, preferably on a fixed schedule, to relieve the pain. Supplemental analgesic medication may also be employed, except that opiate antagonists must be avoided. PMID- 11064493 TI - HIV and HCV infection among injecting drug users. AB - BACKGROUND: Human immunodeficiency virus (HIV) and hepatitis C virus (HCV) are the two blood-borne pathogens most commonly transmitted among injection drug users via multi-person use of syringes and other injection equipment. However, important differences exist in the epidemiology of HIV and HCV within different populations of intravenous drug users. METHOD: A literature review was carried out to summarize publications describing the epidemiology and natural history of HIV and HCV in injection drug users. RESULTS: Among injection drug users worldwide, HIV prevalence varies from <5% to >80%, with annual HIV incidence between <1% and 50%. More consistency is shown in HCV prevalence (50-90%) and incidence (10-30% per year). Host, environmental and viral factors that favor rapid spread of HCV among IDUs suggest that HCV infection in a population of injection drug users may become endemic over a relatively short period of time. Lower transmission efficiency for HIV also indicates that its spread among injection drug users may be somewhat slower. CONCLUSIONS: Successful efforts to prevent transmission of blood-borne viruses among IDUs typically result in risk reduction; however, no intervention has resulted in elimination of risk behavior. To reduce HIV transmission, risk reduction may be sufficient, whereas control of HCV may necessitate the use of injection practices that guarantee elimination of exposure to equipment contaminated with even small amounts of blood. PMID- 11064494 TI - Interactions between methadone and medications used to treat HIV infection: a review. AB - BACKGROUND: It is critical for providers caring for HIV-positive methadone recipients to have accurate information on pharmacologic interactions between methadone and antiretroviral therapy. If providers do not have these data, symptoms of narcotic withdrawal or excess due to medication interactions may be mismanaged, and antiretroviral regimens may be suboptimal in efficacy or associated with increased side effects and toxicities. This review was undertaken to clarify what is known about interactions between pharmacotherapies of opiate dependence and HIV-related medications, to suggest clinically useful approaches to these issues, and to outline areas which need further study. METHOD: A search for relevant published papers and abstracts presented at scientific meetings was conducted using electronic databases. These documents were obtained and reviewed, and additional publications referenced in them were also reviewed. RESULTS: Pharmacokinetic interactions between methadone and zidovudine, didanosine, stavudine, abacavir, nevirapine, efavirenz and nelfinavir have been documented. The mechanisms, clinical implications and management of these interactions are reviewed. CONCLUSIONS: Interactions between methadone and some HIV-related medications are known to occur, yet their characteristics cannot reliably be predicted based on current understanding of metabolic enzyme induction and inhibition, or through in vitro studies. Only carefully designed and conducted pharmacologic studies involving human subjects can help us define the nature of the interactions between methadone (and other pharmacotherapies for opiate dependence) and specific HIV-related medications. Clinicians must be aware of known interactions and be alert to the possibility that interactions which are still undocumented may be present among their patients. PMID- 11064495 TI - The impact of hepatitis C virus infection on methadone maintenance treatment. AB - Hepatitis C virus infection is now recognized as a common and serious complication of injection drug use and will be encountered frequently in methadone maintenance patients. Approximately 1.8% of the United States population, or 3.9 million persons, are infected with hepatitis C virus. A majority of acute hepatitis C virus infections are associated with injection drug use, and 64-88% of injection drug users in seroprevalence studies have antibodies to hepatitis C. Hepatitis C virus infection is almost always chronic, and alcohol use increases the clinical severity. Therapy with interferon and ribavirin will induce long-term remission in up to 43% of patients with hepatitis C virus infection. Proper diagnosis and treatment of hepatitis C virus infection will be indicated for many patients in methadone programs and will require considerable resources. PMID- 11064496 TI - Causes and rates of death among methadone maintenance patients before and after the onset of the HIV/AIDS epidemic. AB - Causes of death and the mortality rates of active methadone patients and those who had left treatment were compared. Prior to the HIV epidemic, death rates among discharged methadone patients were more than twice that of patients who continued with their methadone treatment. However, the death rate from heroin related causes in the post-treatment period was 51 times the rate among active patients. Alcohol-related conditions were the leading causes of death in patients more than 30 years old on methadone. During the post-treatment period, alcohol related deaths were second to those of heroin-related causes. Alcohol-related deaths were particularly pronounced among black patients. Death rates among active male and female patients were identical, but the death rate for discharged female patients was greater than for discharged males. With the onset of the HIV epidemic in the 1980s, AIDS-related causes became the major cause of death in treatment. However, other causes of death, such as alcohol and other medical conditions, identified prior to the AIDS epidemic, persisted. AIDS-related deaths peaked in the mid-1990s and have recently subsided. However, within the past two years, deaths related to HCV have increased to 9% of all patient deaths in a major methadone program. With the emergence of HCV, deaths from this cause are expected to eclipse AIDS-related deaths within the next decade. PMID- 11064497 TI - The converging epidemics of mood-altering-drug use, HIV, HCV, and partner violence: a conundrum for methadone maintenance treatment. AB - BACKGROUND: Accumulating evidence suggests that partner violence may be associated with HIV risk behavior and drug use among women in methadone maintenance treatment programs (MMTPs), yet the mechanisms linking these overlapping problems remain unclear. The main purpose of this qualitative study is to explore in detail how drug-related activities and HIV risk behavior occur in the context of a recent episode of partner violence among women in MMTPs. METHOD: We conducted and analyzed in-depth interviews with 31 women who reported having experienced physical or sexual violence by an intimate partner during the past year. Guided by existing research, feminist theory and trauma theory, we constructed a set of questions which were designed to explore multiple ways in which drug-related activities or HIV risk behavior may be linked directly or indirectly to the recent event. To examine the extent and significance of the woman sand/or her partner s drug-related activities or sexual HIV risk issues occurring immediately before, during and/or after the most recent event, we adapted a series of techniques for thematic analysis of qualitative data. RESULTS: Of the 31 women who reported recent events: 83.8 % (n=26) recalled recent events in which there was some drug-involvement; 40% (n=13) indicated that both she and her partner were involved in drug-related activities during the most recent event of partner abuse; 35% (n=11) reported that the partner was drug involved; and only two women (6.4%) indicated that they alone had been drug involved. One-fifth (19.3%, n=6) of the women indicated that they had used drugs immediately after the event because they were upset or in physical pain. One fifth of the women (n=6) reported that they had coerced, unprotected sex during or after the most recent incident. CONCLUSIONS: The multiple ways in which the use of mood-altering drugs are related to partner violence and the occurrence of coerced, unprotected sex underscore the need to design specific interventions for preventing drug relapse, and HIV and HCV infection among abused women in MMTPs. Treatment and policy implications of study findings are discussed. PMID- 11064498 TI - T-tropic sequence of the V3 loop is critical for HIV-1 infection of CXCR4 positive colonic HT-29 epithelial cells. AB - Some colonic and neuronal cells which are CD4- but galactosyl ceramide-positive are susceptible to infection with HIV-1. We have previously shown that the T-cell tropic V3 loop of HIV-1 gp120 serves as a primary viral determinant for infectivity of CD4- neuronal cells. However, the nature of the V3 loop of HIV-1 needed for infection and the V3 loop's interaction with coreceptors on colonic epithelial cells have not been fully analyzed. By using HIV-1 molecular clones, we show that the T-cell tropic V3 domain is critical for HIV-1 infection of colonic HT-29 epithelial cells. Because T-cell tropic HIV-1 can use CXCR4 as a coreceptor in T cells, we set out to determine the role of CXCR4 during infection of HT-29 cells. Using reverse transcriptase-polymerase chain reaction (RT-PCR) and immunostaining, we show that these epithelial cells of colonic origin express the chemokine receptor CXCR4. Importantly, antibody against CXCR4 or a neutralizing antibody against HIV-1 gp120 V3 loop blocks T-cell tropic HIV-1 entry into HT-29 cells. These data indicate that the V3 loop of HIV-1 and the chemokine receptor CXCR4 are both critical for HIV-1 infection of colonic HT-29 epithelial cells. An HIV-1 T-tropic virus may be responsible for the infection of human colonic epithelial cells in vivo. PMID- 11064499 TI - Antiviral resistance of biologic HIV-2 clones obtained from individuals on nucleoside reverse transcriptase inhibitor therapy. AB - OBJECTIVE: To study phenotypic and genotypic resistance of HIV-2 against nucleoside reverse transcriptase inhibitors (NRTI). METHODS: Biologic HIV-2 clones were generated from 3 patients before and after initiation of antiretroviral therapy with zidovudine (AZT) in patient RH2-7, AZT and didanosine (ddI) in patient PH2-1, and after addition of lamivudine (3TC) to AZT monotherapy in patient RH2-5. The sensitivity to NRTI of the virus clones, as defined by the 50% inhibitory concentration (IC(50)), was determined in vitro. The predicted amino acid sequences of the reverse transcriptase proteins from these clones were determined. RESULTS: Comparing the sensitivity of the biologic HIV-2 clones obtained after start of therapy with those from antiviral naive patients, resistance had developed to AZT (patients RH2-7 and RH2-5) and 3TC (patient PH2-1 and RH2-5). No resistance to AZT was observed in the biologic clone from PH2-1 obtained after start of therapy. The resistant clones from RH2-5 and PH2-1, but not RH2-7, contained amino acid mutations at positions where HIV-1 has been shown to mutate after AZT and 3TC treatment. CONCLUSIONS: Phenotypic resistance of HIV 2 to nucleoside analogues, which developed in HIV-2-infected patients treated with NRTIs, was associated with genotypic changes. Some of the mutations at amino acid positions in the HIV-2 reverse transcriptase gene corresponded with those involved in HIV-1 resistance, although no conventional mutations associated with resistance to AZT were observed. PMID- 11064500 TI - Osteonecrosis in HIV: a case-control study. AB - BACKGROUND: Osteonecrosis (avascular necrosis) has been infrequently reported in HIV-infected patients. It is not known whether HIV itself is an independent risk factor for osteonecrosis. METHODS: We identified 25 patients with osteonecrosis from 1984 to 1999 from a large county teaching hospital and two large practices in Dallas County that specialize in HIV-disease related therapy. A retrospective chart review was performed to evaluate potential risk factors for osteonecrosis. Each case was matched with two controls for HIV positive status and date of osteonecrosis diagnosis. RESULTS: In the study, 22 of 25 (88%) case patients had at least one osteonecrosis risk factor compared with 24 of 50 (48%) controls, p =.003. The most common osteonecrosis risk factors were hyperlipidemia (32%), alcoholism (28%), pancreatitis (16%), corticosteroids (12%), and hypercoaguability (12%). Of the cases, 12% were idiopathic. Multiple joints were involved in 72% of cases. Four of the case patients compared with none of the controls received megesterol acetate before the diagnosis of osteonecrosis, p =.01. No significant differences were found between cases and controls with respect to liver function tests, testosterone levels, triglyceride levels, cholesterol levels, or CD4 cell counts. Saquinavir was independently associated with osteonecrosis, p <.05. However, no differences in overall use of protease inhibitors among cases and controls were noted: 79% versus 76%, respectively. CONCLUSIONS: The increased incidence of osteonecrosis in HIV/AIDS may be due to an increased frequency of risk factors previously associated with osteonecrosis such as hyperlipidemia, corticosteroid use, alcohol abuse, and hypercoaguability. Use of protease inhibitors was not independently associated with osteonecrosis. PMID- 11064501 TI - Comparison of immunologic restoration and virologic response in plasma, tonsillar tissue, and cerebrospinal fluid in HIV-1-infected patients treated with double versus triple antiretroviral therapy in very early stages: The Spanish EARTH-2 Study. Early Anti-Retroviral Therapy Study. AB - The objective of antiretroviral therapy is to obtain an almost complete and durable suppression of viral replication in all compartments to facilitate recovery of the immune system. We assessed the virologic effect in plasma, tonsillar tissue, and cerebrospinal fluid (CSF) in 94 HIV-1-infected patients with CD4 counts >500 x 106 cells per liter and viral load >5000 copies/ml randomly assigned to triple antiretroviral therapy (two nucleoside reverse transcriptase inhibitors (NRTIs) plus one protease inhibitor) versus double therapy (two NRTIs). We also analyzed the immunologic recovery in this cohort of patients. Lymphoid tissue and cerebrospinal fluid viral load, development of genotypic resistance, proliferative responses to HIV-1 specific antigens, and other immunophenotypic markers were analyzed. The proportion of patients who achieved a decrease in HIV RNA levels to <200 copies/ml was significantly greater in the triple therapy group than in the two drug groups (p =.0002 for each pair wise difference). At week 52, tonsillar tissue HIV RNA from 5 patients treated with triple therapy was lower than the limit of detection, whereas the mean +/- standard error in patients with double therapy (n = 5) was 5.03 +/- 0.34 copies/mg/tissue. In all 10 patients, CSF viral load (VL) was <20 HIV-1 RNA copies/ml at week 52. CSF cell counts and protein levels tended to decrease after 52 weeks of antiretroviral therapy. After 1 year of therapy, 13 of 21 patients (62%) in the double-therapy groups (zidovudine plus lamivudine [n = 9] and stavudine plus lamivudine [n = 12]) had evidence of M184V mutation. None of the 10 samples of patients receiving triple therapy could be amplified because of low HIV RNA levels. The mean increase in CD4 cells at week 52 was greater in the stavudine and lamivudine and indinavir group than in the double-treatment arms (186 versus 67 and 102, respectively; p =.03). In patients treated with triple therapy, the increase in naive T cells (CD4 and CD8) was greater than in patients treated with double therapy. Markers of activation decreased further in patients treated with the regimen that included protease inhibitors. Proliferative responses to HIV-1 p24 antigen were never recovered after double or triple therapy. Our study suggests that even in very early stages of HIV-1 disease only therapy with two NRTIs and one protease inhibitor reduces plasma, lymphoid tissue, and CSF VL to undetectable levels. HIV-1-related immune system abnormalities improved but were still defective after 1 year of antiretroviral therapy. PMID- 11064502 TI - Duration of viral suppression in patients on stable therapy for HIV-1 infection is predicted by plasma HIV RNA level after 1 month of treatment. AB - The aim of this study was to assess the predictive value of HIV RNA levels after 1 month of therapy on the long-term virologic outcome in an unselected general population of HIV-infected patients. DESIGN: Analysis was conducted retrospectively on an ongoing clinical cohort of HIV-positive patients who were receiving antiretroviral treatment. Data on 575 patients were analyzed. RESULTS: The HIV RNA value at 1 month was significantly correlated with the virologic outcome after 12 and 24 months of therapy (R = 0.258 and R = 0.44, respectively). The predictive value of the 1-month viral load was also statistically significant after stratification for baseline CD4 T-cell counts. Prediction was similar in highly compromised patients (CD4 < or = 100 cells/microl; R = 0.426; p = .001) or in patients with a better immunologic status (R = 0.419; p < .0001). It retained validity in patients who were naive or experienced for antiretroviral therapy. CONCLUSION: HIV RNA level after 1 month of therapy is a useful prognostic marker in HIV-infected patients. It predicts long-term virologic and immunologic outcome. A cutoff level of 5000 copies/ml identifies patients most likely to fail current therapy. In these patients, a more aggressive strategy or specific diagnostic interventions to clarify the relative influence of viral resistance and/or subtherapeutic regimens is advised. PMID- 11064503 TI - Oral mucosal lesions and HIV viral load in the Women's Interagency HIV Study (WIHS). AB - The prevalence of oral lesions was assessed in a five-center subset of the Women's Interagency HIV Study (WIHS) and correlated with other features of HIV disease. Oral examinations were performed by dental examiners on 729 women (577 HIV-positive and 152 HIV-negative) during baseline examination. Significant differences between the groups were found for the following oral lesions: pseudomembranous candidiasis, 6.1% and 2.0%, respectively; erythematous candidiasis, 6.41% and 0.7%, respectively; all oral candidiasis, pseudomembranous and/or erythematous, 13.7% and 3.3%, respectively. Hairy leukoplakia was observed in 6.1% of HIV-positive women. No significant differences were found for recurrent aphthous ulcers, herpes simplex lesions, or papillomas. Kaposi's sarcoma was seen in 0.5% of HIV-positive and 0% of HIV-negative women. Using multiple logistic regression models controlling for use of antiretrovirals and antifungals, in HIV-positive women the presence of oral candidiasis was associated with a CD4 count <200 cells/microl, cigarette smoking, and heroin/methadone use; the presence of hairy leukoplakia was not related to CD4 count but was associated with high viral load. Oral candidiasis and hairy leukoplakia are confirmed as being common features of HIV infection in women and appear to be associated with HIV viral load, immunosuppression, and various other behaviorally determined variables. PMID- 11064504 TI - Care of vaginal symptoms among HIV-infected women. AB - OBJECTIVE: Gynecologic disease is common in HIV-infected women. We examine the sociodemographic, clinical, and provider factors associated with the care of women with vaginal symptoms. METHODS: Women enrolled in the HIV Cost and Services Utilization Study (HCSUS), a nationally representative probability sample of HIV infected adults, were interviewed between January 1996 and April 1997. Women with vaginal symptoms who sought medical attention were asked, "Did your health care provider examine your vaginal area?" Women were also asked if they received medication for their symptoms. RESULTS: Among 154 women with vaginal symptoms, 127 sought care for their symptoms. Of those who sought care, 48% saw a gynecologist and 52% sought care from nongynecologists, most often their usual HIV care provider. Women who saw a gynecologist for their symptoms were more likely to have received a pelvic examination (92% versus 76%; p =.06) and vaginal fluid collection (98% versus 88%; p =.06) than those who saw their regular HIV provider. Fifteen percent of women received medication for their symptoms without having a pelvic examination; gynecologists were less likely to prescribe without an examination (8% versus 21%; p =.12). CONCLUSION: Gynecologists are more likely to provide adequate care of vaginal symptoms among HIV-infected women than nongynecologists who were HIV care providers. This specialty difference is consistent with quality of care studies for other medical conditions, but the potential gynecologic complications of inadequate evaluation and treatment warrants further investigation. PMID- 11064505 TI - Plasma viral load concentrations in women and men from different exposure categories and with known duration of HIV infection. I.CO.N.A. Study Group. AB - CONTEXT: According to recent studies, women have lower plasma HIV RNA concentrations than men. However, these studies did not take into account the duration of HIV infection. OBJECTIVES: To analyze the relationship between viral load and gender among individuals with known date of seroconversion. SETTING: Sixty infectious disease clinics in Italy. DESIGN: Cross-sectional analysis of data collected at enrollment in a cohort study. PARTICIPANTS: Injecting drug users and heterosexual contacts naive to antiretroviral therapy at enrollment (245 men; 170 women). MAIN OUTCOME MEASURES: Plasma HIV RNA concentrations, measured using quantitative reverse transcriptase-polymerase chain reaction (RT PCR) or signal amplification b-DNA assays before antiretroviral therapy. RESULTS: Plasma HIV RNA concentrations were similar by age and exposure category (p =.80 and p =.39, respectively). Median viral load among women was roughly half that of men (p =.002). The association between viral load and gender remained significant after fitting a two-way analysis of variance (p =.03) and after adjusting for CD4 count, modality of HIV transmission, and age at enrollment in a regression model. Viral load was 0.27 log10 copies/ml (95% confidence interval, 0.05-0.40; p =.01) lower in women (i.e., 50% lower in the raw scale). CONCLUSIONS: Plasma HIV RNA concentrations were found to be lower among women, even when considering the duration of HIV infection. Compared with men, it is possible women should be given highly aggressive antiretroviral therapy at lower HIV-RNA concentrations. PMID- 11064506 TI - Prevention of HIV infection in street-recruited injection drug users. The Collaborative Injection Drug User Study (CIDUS). AB - BACKGROUND: Injection drug users (IDUs) and their sex partners account for an increasing proportion of new AIDS and HIV cases in the United States, but public debate and policy regarding the effectiveness of various HIV prevention programs for them must cite data from other countries, from non-street-recruited IDUs already in treatment, or other programs, and from infection rates for pathogens other than HIV. METHODS: Participants were recruited from the street at six sites (Baltimore [Maryland], New York [two sites], Chicago [Illinois], San Jose [California], Los Angeles [California], and at a state women's correctional facility [Connecticut]), interviewed with a standard questionnaire, and located and reinterviewed at one or more follow-up visits (mean, 7.8 months later). HIV serostatus and participation in various programs and behaviors that could reduce HIV infection risk were determined at each visit. RESULTS: In all, 3773 participants were recruited from the street, and 2306 (61%) were located and interviewed subsequently. Of 3562 initial serum specimens, 520 (14.6%) were HIV seropositive; at subsequent assessment, 19 people, all from the East Coast and Chicago, had acquired HIV. Not using previously used needles was substantially protective against HIV acquisition (relative risk [RR], 0.29; 95% confidence interval [CI], 0.11-0.80 ) and, in a multivariate model, was significantly associated with use of needle and syringe exchange programs (adjusted odds ratio [ORadj], 2.08; 95% CI, 1.15-3.85). Similarly, reduction of injection frequency was very protective against seroconversion (RR, 0.33; 95% CI, 0.14-0.80), and this behavior was strongly associated with participation in drug treatment programs (ORadj, 3.54; 95% CI, 2.50-5.00). In a separate analysis, only 37.5% of study-participants had sufficient new needles to meet their monthly demand. CONCLUSIONS: In this large multicity study of IDUs in the United States, several HIV prevention strategies appeared to be individually and partially effective; these results indicate the continued need for, and substantial gaps in, effective approaches to preventing HIV infection in drug users. PMID- 11064507 TI - Increasing proportion of late diagnosis of HIV infection among patients with AIDS in Italy following introduction of combination antiretroviral therapy. AB - We analyzed trends over time and determinants of late diagnosis of HIV infection among people diagnosed with AIDS in 1986 to 1998 in a tertiary care center in Rome, Italy. Information on the date of a first HIV test was collected prospectively, in addition to data routinely collected for AIDS reporting. Patients with AIDS were defined as "late testers" if the time interval between first positive HIV test result and AIDS diagnosis was < or = 3 months. Overall, 503 people with AIDS of 1977 included in the analysis (25.4%) were late testers. the proportion of late testers decreased from 62.5% in 1986 to 16% in 1995. Thereafter, this proportion increased to 20.5% in 1996, 33.7% in 1997, and 36.6% in 1998. In multivariate analysis, the following variables were significantly associated with late testing: AIDS diagnosis in years 1986 to 1993 or 1997 to 1998 compared with 1995, male gender, age > or = 45 years, men who have sex with men, heterosexual contacts, or having unknown transmission mode compared with intravenous drug users, and being born outside Italy. Since 1996, the overall number of AIDS cases diagnosed at our center began to decrease whereas the number of late-testing AIDS patients did not decrease, resulting in an increasing proportion of late testers during the last 3 years of the study. This findings may reflect the effect of combination antiretroviral therapy in slowing progression to AIDS of HIV-infected persons aware of their status. A relevant number of people still discover their HIV infection late and may therefore miss treatment opportunities. New testing strategies are needed to reach more people who engage in high-risk behaviors, especially those at risk for sexual transmission, and those born outside Italy. PMID- 11064508 TI - Looking for change in response to the AIDS epidemic: trends in AIDS knowledge and sexual behavior in Zambia, 1990 through 1998. AB - This study investigates trends in AIDS knowledge and sexual behavior among men and women in urban Lusaka 1990 to 1998, and in all of Zambia, 1992 to 1998. Using data from representative surveys of urban Lusaka and of the country as a whole, population proportions were estimated to examine trends in knowledge and sexual risk behaviors. Differences in the estimated proportions between 1990 and 1998 were tested in Lusaka. In all Zambia, tests of difference were conducted between the earliest and latest years for which data were available for each indicator. A decline in premarital sexual activity was observed in urban Lusaka. In 1990, 50% of never married women reported no sexual experience, compared with 60% in 1998 (p =. 003); among men, the figures were 38% and 53%, respectively (p <. 001). Fewer women (1990, 8%; 1998, 2%; p <.001) and men (1990, 31%; 1998, 19%; p =.07) had extramarital partners. The bulk of change observed in urban Lusaka took place from 1990 to 1996; the changes in men's behavior observed between 1996 and 1998 were also observed in the national estimates for those years. National figures for other indicators from 1992 to 1998 were less encouraging. Apart from an increase in having ever used condoms, no change in women's sexual behavior was observed. Fewer men had premarital sex from 1996 to 1998 (1996, 64%; 1998, 46%; p <.001), but condom use with nonregular partners decreased among men (1996, 38%; 1998, 29%; p =.02). Prevention campaigns focused on education about AIDS and promoting safer sexual behavior appear to have made a difference in the early 1990s in Zambia. Findings from more recent years indicate that further change has stagnated. Renewed efforts are needed, particularly targeting condom use with nonregular partners. PMID- 11064509 TI - The association between cocaine use and HIV/STDs among soup kitchen attendees in New York City. AB - We examined the associations of cocaine use with HIV/sexually transmitted diseases (STDs) in a sample of 184 soup kitchen attendees using a mobile medical van in Manhattan (male = 66%; black or Hispanic = 81%; cocaine use, primarily crack = 75%; ever injected drugs = 22%). In addition to confirming the association between years of cocaine use and HIV antibodies in this sample (odds ratio [OR] = 2.11; p <.05) we examined the pattern of associations of cocaine use and non-HIV STDs under the hypothesis that the strength of an association depends on the efficiency of sexually transmitting a particular STD (high, moderate, and low for syphilis, hepatitis B and hepatitis C, respectively). As predicted, years of cocaine use was strongly associated with syphilis (OR = 2.07; p <.05), moderately associated with hepatitis B core antibodies (OR = 1.50; p <.05), and not significantly associated with hepatitis C antibodies (OR = 1.48; p >.05). A reverse pattern of associations between opiate use (injection drug use) and the three STDs points to the singular significance of cocaine use in the sexual transmission of STDs, and by inference, HIV. This conclusion is further bolstered by correlations of biologic (hair assays) and self-reported measurements of cocaine use (but not opiates) with self-reports of high risk sexual behavior among the women (number of partners and selling sex) and men (number of partners and buying sex). These data underscore the need for effective cocaine treatment and HIV interventions tailored to the large numbers of cocaine users in inner cities. PMID- 11064510 TI - Does alcohol intake affect highly active antiretroviral therapy (HAART) response in HIV-positive patients? PMID- 11064511 TI - Nevirapine plus didanosine: once or twice daily combination? PMID- 11064512 TI - Prevalence of novel lamivudine-resistant genotypes (E44D/A, V118I) in naive and pretreated HIV-infected individuals. PMID- 11064513 TI - Lipodystrophy and nucleoside analogue therapy in HIV-infected patients: important question, few valid answers. PMID- 11064514 TI - Per-exposure rate of transmission of HIV-1, HIV-1/2, and HIV-2 from women to men may be higher in India. PMID- 11064515 TI - The visible human anatomy of the lumbar erector spinae. AB - STUDY DESIGN: Image data of the male and female cadavers from the Visible Human Project were visualized and quantified. OBJECTIVE: To clarify the anatomy of the lumbar part of the human lumbar erector spinae muscles. SUMMARY OF BACKGROUND DATA: Recent studies have shown discrepancies in the description of the anatomy of the lumbar part of the lumbar erector spinae. The main differences concern whether lumbar fascicles of iliocostalis lumborum exist and whether the lumbar fascicles have direct attachments to the ilium or attach via the erector spinae aponeurosis. With the Visible Human Project from the U.S. National Library of Medicine, a new powerful basis for anatomic investigation has become available. METHODS: Software was produced to visualize sections oriented in any direction and with maximum resolution of the Visible Human male and female. Three dimensional coordinates of anatomic structures in the image space could be marked in the cross-sectional images. The geometry and the physiologic cross-sectional areas of the erector spinae fascicles of lumbar origin were thus derived. RESULTS AND CONCLUSIONS: The study supports a classification of the lateral fascicles of the lumbar part of the lumbar erector spinae as part of iliocostalis lumborum. In both the male and the female, a large part of the erector spinae fibers of lumbar origin attached to the erector spinae aponeurosis. These results are of importance for biomechanical analysis of force transmission in the lumbar spine. PMID- 11064516 TI - Pedicle morphology of the immature thoracolumbar spine. AB - STUDY DESIGN: Human vertebral morphologic data were compiled from anatomic skeletal collections from three museums. OBJECTIVES: To quantify the morphometric characteristics of the pedicles of the immature thoracolumbar spine. SUMMARY OF BACKGROUND DATA: Little is known of pedicle morphology of the immature spine as related to pedicle screw fixation. METHODS: A total of 75 anatomic skeletal specimens were acquired from C1 to L5 in the age range of 3 to 19 years. The data were collected and analyzed using a computerized video analysis system. Each vertebral pedicle was measured in the axial and sagittal planes. The measurements included the minimum pedicle width, the pedicle angle, the distance to anterior cortex, and anteroposterior and interpedicular spinal canal diameters. RESULTS: Wide variation in pedicle morphology between specimens at each vertebral level was found in the young population. In general, compared with the average adult data, a younger spine demonstrated a near uniform reduction in the linear pedicle dimensions at each vertebral level. Pedicles from the lower lumbar vertebrae attained dimensions acceptable for standard screw sizes at an earlier age than in the thoracic vertebrae. CONCLUSIONS: The data in this study indicates that pedicle screws may be used in the adolescent spine. However, care should taken to accurately ascertain pedicle size before surgery so that improper use of screws can be avoided. Growth of the pedicles in relation to the spinal canal indicates that the increase in pedicle size is lateral to the spinal canal. PMID- 11064517 TI - Observations on fiber-forming collagens in the anulus fibrosus. AB - STUDY DESIGN: The spatial distribution of fiber-forming collagens in the anulus fibrosus was investigated in the complete longitudinal and horizontal sections of human lumbar intervertebral discs of seven individuals. OBJECTIVES: To obtain a more detailed structural definition of the anulus fibrosus because structural alterations of its collagen fiber network have been implicated in discal degeneration and other spinal pathologies. SUMMARY OF BACKGROUND DATA: Prior biochemical or immunofluorescence studies permitted only limited conclusions concerning the spatial distribution of the fiber-forming collagens in relation to anatomic structures because they were based on intraoperative tissue specimens or performed on incomplete sections of human intervertebral discs. METHODS: Complete human intervertebral discs with their adjacent vertebral bodies were fixed, decalcified, and embedded in paraffin. The intervertebral disc and its adjacent structures were reviewed in their entirety on one histologic slide. Monoclonal antibodies against human Types I, II, and III collagen were used for immunohistochemistry. A comparative analysis based on both immunohistochemical and histologic evaluation was performed. RESULTS: Type I collagen was seen abundantly in the outer zone and outer lamellas of the inner zone of the anulus fibrosus. On longitudinal sections, the Type I collagen distribution took the shape of a wedge. On horizontal sections, the Type I collagen positive area took the shape of a ring that was wider anteriorly than posteriorly. This suggests that the three-dimensional shape of the Type I collagen-positive tissue in the anulus fibrosus can be described by a donut that is wider anteriorly than posteriorly. Type II collagen was present in the entire inner of the anulus fibrosus, but not in the outer zone. In addition, it was found in the cartilaginous endplates. Type III collagen showed some codistribution with Type II collagen, particularly in pericellular locations in areas of spondylosis, which was noted at the endplates, vertebral rim, and insertion sites of the anulus fibrosus. CONCLUSIONS: These observations on the location of Types I and II collagen provide a more detailed structural definition of the anulus fibrosus, which may assist in further investigation of discal herniation. PMID- 11064518 TI - Fibronectin and its fragments increase with degeneration in the human intervertebral disc. AB - STUDY DESIGN: This laboratory-based experiment correlates fibronectin content of intervertebral disc with a morphologic grade of degeneration. OBJECTIVES: To correlate the fibronectin content of the anulus fibrosus and nucleus pulposus with a gross morphologic grade of disc degeneration, and to determine the molecular size of the extractable fibronectin. SUMMARY OF BACKGROUND DATA: Intervertebral disc degeneration increases with age and can lead to low back pain. Fibronectin helps to organize the extracellular matrix and provides environmental cues by interaction with cell surface integrins. In other tissues, its synthesis is elevated in response to injury. Fibronectin fragments can stimulate cells to produce metalloproteases and cytokines and inhibit matrix synthesis. METHODS: In this study, 17 anuli fibrosis and 18 nuclei pulposus from 11 spines were graded by Thompson's gross morphologic scale. Fibronectin was sequentially extracted with 4 mol/L guanidine hydrochloride and trypsin, and then quantitated by enzyme-linked immunoassay. The size of extractable fibronectin was determined by Western blot analyses. RESULTS: The fibronectin content of the disc increased with grade and was significantly elevated between Grades 3 and 4. The percentage of extractable fibronectin varied widely, but it was more extractable from the nucleus. In both the nucleus and anulus, 30% to 40% of the extractable fibronectin existed as fragments. Many of the fragments contained functional heparin or collagen-binding sites. CONCLUSIONS: Fibronectin is elevated in degenerated discs and frequently present as fragments. Elevated levels of fibronectin suggest that disc cells are responding to the altered environment. Fibronectin fragments resulting from normal or enhanced proteolytic activity could be a mechanism that induces the cell to degrade the matrix further. PMID- 11064519 TI - A study of stiffness protocol as exemplified by testing of a burst fracture model in sagittal plane. AB - STUDY DESIGN: An in vitro biomechanical study of lumbar spine segments. OBJECTIVE: To study the characteristics of the stiffness test protocol. SUMMARY OF BACKGROUND DATA: In an in vitro study using a flexibility protocol, forces are applied and motions are measured; no center of rotation needs to be specified. In a study using a stiffness protocol, the forces are measured and the motions are applied. This does require the center of rotation to be specified. Many biomechanical studies of the spine are available, but there is lack of clarity concerning which of these two test protocols is appropriate to achieve a certain study goal. METHODS: Five-vertebrae lumbar spine specimens with burst fractures in the middle vertebrae (L1) were used. Specially designed apparatus applied flexion and extension rotations around five centers of rotations located on anteroposterior line through the middle of L1. Maximum moment of 4 Nm was applied. RESULTS: The authors found load-displacement curves, ranges of motion, and neutral zones obtained at the five centers of rotations to be markedly different. The center of rotation located at the posterior longitudinal ligament produced large range of motion and neutral zones in comparison to the centers of rotation located at the anterior longitudinal ligament and the spinous process tip (P<0.01). CONCLUSIONS: The stiffness protocol requires that a center of rotation be specified. Shown here is the significant variability in the load displacement curves, depending on the choice of the location of the center of rotation. Certain center of rotation locations may block the natural motions of the spine and may result in tissue damage. PMID- 11064520 TI - Anterior vertebral screw strain with and without solid interspace support. AB - STUDY DESIGN: This in vitro biomechanical study examines segmental anterior vertebral screw strain and solid rod construct stiffness with and without the addition of multilevel, threaded cortical bone dowels in a bovine model. OBJECTIVE: To determine whether strain at the bone-screw interface is higher at the end levels during physiologic range loading, and whether solid interspace support decreases segmental strain on the implant. SUMMARY OF BACKGROUND DATA: Anterior instrumentation provides greater correction and preserves distal motion segments. However, nonunion and implant failure are observed more frequently than with posterior segmental instrumentation, and when observed, loss of fixation occurs at the end levels. METHODS: Eight calf spines underwent mechanical testing in the following sequence: 1) intact condition, 2) anterior release with anterior solid rod and bicortical rib grafts, and 3) anterior release with anterior solid rod and threaded cortical bone dowels (L2-L5). Instrumented vertebral screws were used to assess strain within the vertebral body by the near cortex, whereas an anterior extensometer spanning the instrumented segments was used to measure segmental displacements to calculate construct stiffness. The protocol included axial compression (-400 N), right lateral bending (4 Nm (Newton-meter), away from the implant), and left lateral bending (4 Nm, toward the implant). Statistical analysis included a one-way analysis of variance and a Student-Newman-Keuls post hoc test. A pilot study was performed using four additional specimens loaded for 4000 cycles to investigate macroscopic loosening after fatigue loading. RESULTS: In lateral bending toward the implant, the strain was higher at both end levels, with no differences between the rib and dowel reconstructions. The stiffness values were greater than the intact values for both groups. In lateral bending away from the implant, the strain also was higher at both end screws, and the dowel group had less strain at these levels than the rib group. Both groups were stiffer than the intact condition, and the dowel group was stiffer than the rib group. Axial compressive strain also was higher at the end levels, but this difference did not reach statistical significance. The rib group did not reach intact stiffness values, whereas the dowel group was stiffer than the intact condition. The fatigue study showed gross loosening at one or both end levels in all cases. CONCLUSIONS: Higher strain was observed at the bone-screw interface in both end screws of an anterior solid rod construct during lateral bending, which correlates with the clinically observed failure location. This suggests that physiologic range loading may predispose to failure at the end levels. Disc space augmentation with solid implants increased construct stiffness in all three load paths and decreased strain at the end levels in lateral bending away from the implant. Future implant modifications should achieve better fixation at the end screws, and the current model provides a means to compare different strategies to decrease strain at these levels. PMID- 11064521 TI - Subsidence resulting from simulated postoperative neck movements: an in vitro investigation with a new cervical fusion cage. AB - STUDY DESIGN: A biomechanical in vitro subsidence test of different cervical interbody fusion devices was performed using a new testing protocol that simulates physiologic conditions. OBJECTIVES: To investigate the effect of simulated postoperative neck movements on the subsidence of the new WING cervical interbody fusion cage in comparison with two other cages and bone cement. SUMMARY OF BACKGROUND DATA: Cervical interbody fusion cages sometimes cause complications because of subsidence into the adjacent vertebrae with collapse of the intervertebral space. Complications such as cage dislocation or nonunion with instability also have been reported. To prevent such complications, the new WING cervical interbody fusion cage (Medinorm AG, Quierschied, Germany) has been developed. Its area of contact with the adjacent vertebrae is supposed to be large enough to resist excessive subsidence and small enough to prevent stress protection of the tissue growing in the cage. METHODS: In this study, 24 human cervical spine specimens were tested after stabilization with either a WING, BAK/C, AcroMed I/F cage or bone cement. Then, in a new testing protocol, 700 pure moment loading cycles (+/-2 Nm) were applied in randomized directions (lateral bending, flexion-extension, and axial rotation alone or in combination with each other) to simulate the patient's neck movements during the first few postoperative days. Measurements of the subsidence depth (total height loss) in combination with flexibility tests (+/-2.5 Nm) were performed before cyclic loading and after 50, 100, 200, 300, 500, and 700 loading cycles. RESULTS: Cyclic loading caused subsidence in all four device groups, most distinct with BAK/C cages (1.63 mm after 700 loading cycles) followed by the new WING (0.90 mm) and the AcroMed (0.82 mm) cages. No statistically significant difference could be found among the three cage designs. However, all three cage types showed a significantly higher subsidence depth than bone cement (0.48 mm;P = 0.023 between each of the three cage-types and bone cement). A moderate correlation between bone mineral density and subsidence depth could be found only in the BAK/C group (r2 = 0.495). A large subsidence depth after 700 loading cycles was associated with a large flexibility increase in the WING (r2 = 0.786) and AcroMed groups (r2 = 0.21), but with a small flexibility increase in the BAK/C group (r2 = 0.58). CONCLUSIONS: Postoperative neck movements caused subsidence in all cervical interbody implant types. The new WING cage and the AcroMed cage seemed to have a better resistance against subsidence than the BAK/C cage. However, all three cage types had a significantly higher subsidence tendency than bone cement. PMID- 11064522 TI - Increase in spinal canal area after inverse laminoplasty: an anatomical study. AB - STUDY DESIGN: In vitro measurement of the area of the spinal canal in the rostral and caudal portions of lumbar vertebrae before and after application of a new technique called "inverse laminoplasty." OBJECTIVES: To quantify the normal area of the spinal canal in the rostral and caudal portions of lumbar vertebrae and the amount of enlargement gained after inverse laminoplasty. SUMMARY AND BACKGROUND DATA: Other types of laminoplasty have been proven to increase the area of the spinal canal. Inverse laminoplasty has been performed in 10 patients but has not been evaluated in vitro. METHODS: The transverse and anteroposterior diameter of the spinal canal was measured in 34 vertebrae from seven cadavers using digital calipers. In each vertebra, the laminae and spinous process were removed en bloc using a high-speed drill. The removed piece was inverted and reattached with titanium mini-plates. The area of the spinal canal was again measured and compared with the prelaminoplasty measurements using paired Student's t tests. RESULTS: The anteroposterior diameter and area of the spinal canal were significantly smaller before surgery in the rostral than in the caudal part of the vertebrae (P <10(-3)). The rostral and caudal areas of the spinal canal increased by 61% and 17%, respectively, after the laminae were inverted (P <10(-3)). CONCLUSION: Because inverse laminoplasty is simple and increases the area of the spinal canal, it may prove to be a useful surgical technique for the treatment of lumbar spinal stenosis. Further studies are needed to determine whether the technique is biomechanically sound and whether it helps prevent perineural scarring. PMID- 11064523 TI - Retro-dental reactive lesions related to development of myelopathy in patients with atlantoaxial instability secondary to Os odontoideum. AB - STUDY DESIGN: A retrospective analysis of 13 patients with atlantoaxial instability secondary to Os odontoideum who underwent posterior atlantoaxial fusion. OBJECTIVE: To assess the relationships between the development of myelopathy and plain radiographic parameters in patients with atlantoaxial instability secondary to Os odontoideum and to determine whether the pathologic structures, which compress the spinal cord, are visualized using magnetic resonance imaging. SUMMARY OF BACKGROUND DATA: The development of myelopathy, which is the most serious complication associated with Os odontoideum, was thought to be related to either the degree of instability or direction of instability, or a decrease in the space available for the cord. However, such indirect radiographic parameters measured using plain radiographs cannot provide direct information concerning the causes of myelopathy in patients with atlantoaxial instability secondary to Os odontoideum. METHODS: Thirteen patients who underwent posterior atlantoaxial fusion for clinical symptoms due to Os odontoideum were classified into two groups depending on whether they had (n = 9) or did not have (n = 4) myelopathy. Four radiographic parameters were measured using flexion and extension lateral radiographs; the degree of instability, the direction of instability, and the space available for the cord in flexion and extension. MRI was performed on all patients in the myelopathy group. The radiologic and clinical data were compared for the two groups. RESULTS: There were no significant statistical differences in the degree of instability (6.83 vs. 7.38, P = 0.816), space available for the cord in flexion (6.94 vs. 7.13, P = 0.938), and space available for cord in extension (7.56 vs. 5.75, P = 0.434) between the two groups. There was a poor agreement between the direction of instability and the development of myelopathy (kappa = 0.268, P = 0.308). Magnetic resonance imaging did demonstrate, however, cord compression caused by retro-dental reactive lesions in the myelopathy; cystic masses were present in two patients; and fibrocartilaginous masses were present in seven. CONCLUSION: The current study suggests that the value of plain radiographic parameters should be reevaluated as a means of evaluating myelopathy in patients with atlantoaxial instability secondary to Os odontoideum, and that retro-dental reactive lesions should be considered as the potential cause of myelopathy. PMID- 11064524 TI - Exercise therapy for low back pain: a systematic review within the framework of the cochrane collaboration back review group. AB - STUDY DESIGN: A systematic review of randomized controlled trials was performed. SUMMARY OF BACKGROUND DATA: Exercise therapy is a widely used treatment for low back pain. OBJECTIVES: To evaluate the effectiveness of exercise therapy for low back pain with regard to pain intensity, functional status, overall improvement, and return to work. METHODS: The Cochrane Controlled Trials Register, Medline, Embase, PsycLIT, and reference lists of articles were searched. Randomized trials testing all types of exercise therapy for subjects with nonspecific low back pain with or without radiation into the legs were included. Two reviewers independently extracted data and assessed trial quality. Because trials were considered heterogeneous with regard to study populations, interventions, and outcomes, it was decided not to perform a meta-analysis, but to summarize the results using a rating system of four levels of evidence: strong, moderate, limited, or none. RESULTS: In this review, 39 trials were identified. There is strong evidence that exercise therapy is not more effective for acute low back pain than inactive or other active treatments with which it has been compared. There is conflicting evidence on the effectiveness of exercise therapy compared with inactive treatments for chronic low back pain. Exercise therapy was more effective than usual care by the general practitioner and just as effective as conventional physiotherapy for chronic low back pain. CONCLUSIONS: The evidence summarized in this systematic review does not indicate that specific exercises are effective for the treatment of acute low back pain. Exercises may be helpful for patients with chronic low back pain to increase return to normal daily activities and work. PMID- 11064525 TI - Osseous anatomy of the lumbosacral spine in Marfan syndrome. AB - STUDY DESIGN: This study examines pedicle widths, laminar thicknesses, and scalloping values for lumbosacral spine elements in Marfan volunteers. Comparisons were made between these measurements and norms as well as measurements between Marfan patients with and without dural ectasia. OBJECTIVES: To determine if the lumbosacral vertebral elements are altered in the patient with Marfan syndrome. SUMMARY OF BACKGROUND DATA: Several abnormalities have been noted in Marfan lumbar spine, including pedicular attenuation and widened interpediculate distances. This may be due to abnormalities of growth or presence of dural ectasia. Given the large numbers of Marfan patients requiring spinal surgery and the high postoperative failure rate, better understanding of the bony anatomy of Marfan lumbar spine is necessary, especially if use of instrumentation is anticipated. METHODS: Thirty-two volunteers with Marfan syndrome based on the Ghent criteria underwent spiral computed tomography of the lumbosacral spine. Images were evaluated for dural ectasia, and measurements of pedicle width, laminar thickness, and vertebral scalloping were made. RESULTS: Pedicle widths and laminar thicknesses were significantly smaller in Marfan patients at all levels (P<0.001). Mean pedicle widths at L1-L3 were smaller than the smallest available pedicle screw (5 mm). In Marfan patients with dural ectasia, laminar thickness from L5-S2 and pedicle widths at all lumbar levels were significantly reduced (P<0.01). Vertebral scalloping at S1 was significantly greater in Marfan patients with dural ectasia (P = 0.02). CONCLUSION: Lumbar pedicle width and laminar thickness are significantly reduced in Marfan individuals. Those with dural ectasia demonstrate increased bony erosion of anterior and posterior elements of lumbosacral spine. Preoperative planning and routine computed tomography scans are recommended when operating on Marfan lumbosacral spine. PMID- 11064526 TI - Inflammatory cells, motor weakness, and straight leg raising in transligamentous disc herniations. AB - STUDY DESIGN: Possible statistically significant relationships between inflammatory cells and either motor weakness or straight leg raising were determined. OBJECTIVES: To look for any clinically relevant links between inflammatory cells in disc herniations and signs of radiculopathy. SUMMARY OF BACKGROUND DATA: Many studies have during recent years shown a presence of various types of inflammatory cells in disc herniations, but their clinical relevance has been questioned. To be clinically relevant, a presence of inflammatory cells should show a clear relationship to clinical evidence of nerve root involvement. Macrophages repeatedly demonstrated in a high proportion of disc herniations studied are of particular interest. Their major role may be in disc herniations tissue resorption and not in sciatica. METHODS: A total of 96 disc herniations, all transligamentous, were analyzed by immunohistochemistry for presence of macrophages, T or B lymphocytes, and activated T lymphocytes separately. From recorded patient data, motor weakness and straight leg raising data were compared with a presence or absence of abundant (+ = at least 20 cells in a group) inflammatory cells. When not abundant, inflammatory cells were classified as "only few cells" (+) and grouped together with "no cells" (-). Patients with or without motor weakness were compared. Straight leg raising was compared for a positive (at <70 degrees ) or a negative test, and separately using the median as cut-off value. Groups were compared by chi-square analysis with the level of statistical significance set at P<0.05. RESULTS: None of the four inflammatory cell types showed any significant association with motor weakness. Nor was any association observed when comparing positive and negative straight leg raising. With the median (straight leg raising = 47.5 degrees ) as cut-off, only activated T cells showed a weak (chi2 = 4.40, P<0.05) relationship with tighter straight leg raising, but none of the other cell types did. Even when straight leg raising was < 47.5 degrees, three times more disc herniations lacked (n = 34) inflammatory cells than showed (n = 13) inflammation. In a subgroup of only sequestrated discs, the findings were similar. However, in the patients with a bilaterally positive straight leg raising (n = 25), the prevalence of at least one inflammatory cell type was much higher in sequestrated discs (80%) than in extrusions (33%). This may suggest more subtle interrelationships between type of disc herniation, straight leg raising, and inflammatory cells. CONCLUSIONS: The results of this study do not support a clinically relevant role for disc herniation inflammatory cells in sciatica. For the cells to be clinically relevant, a strong relationship between a presence of inflammatory cells and either or both of motor weakness and a tight straight leg raising should have been observed. The authors conclude that macrophages, which have been demonstrated in a high proportion of disc herniations in previous studies, are probably more important for disc tissue resorption processes than for producing sciatica. Other types of inflammatory cells are more rarely observed and may have no clinical meaning at all. However, more subtle interrelationships, considering the various types of disc herniations, should be further explored. PMID- 11064527 TI - Congruent spinopelvic alignment on standing lateral radiographs of adult volunteers. AB - STUDY DESIGN AND OBJECTIVES: Radiographs of 75 healthy volunteers were measured to decide parameters and ranges for "congruent" sagittal spinopelvic alignments using the pelvic radius technique. A subset of 30 of the volunteers subsequently had a second radiograph to assess for changes in the repeated measurements. SUMMARY OF BACKGROUND DATA: Measurement of spinal alignment is important. Radiographic parameters for "congruent" spinopelvic balance over the hips and changes in sagittal spinal alignments over time have not been defined. Measurement techniques for spinal alignments and to quantitate pelvic morphology need to be standardized. METHODS: The 75 volunteers (44 men/31 women, mean age 39 years, range, 20 to 63 years) had 36-inch standing lateral radiographs of the thoracolumbar spine and pelvis taken that included both hips. Thirty volunteers (19 men/11 women) had a second radiograph taken 5 to 6 years later. Radiographic measurements were made using the pelvic radius technique. This required locating a midpoint between the approximate centers of both femoral heads to establish a pelvic hip axis. A line between the hip axis and the posterior superior corner of S1 for the pelvic radius was drawn and measured for length. Angles were measured from the pelvic radius to tangents along the vertebral endplates on the 105 films with an electronic digital readout device. These angles included PR-S1 for pelvic morphology and PR-T12 for total lumbopelvic lordosis. A pelvic angle was measured from a vertical line through the hip axis to the pelvic radius. This angle gave the sagittal alignment for the pelvis over the hips. Longitudinal measurements between radiographs were compared for minimum and maximum change. Significant statistical correlations for the measurements were carefully studied to determine potentially important clinical relationships. In addition, thoracic kyphosis/lumbar lordosis ratios were assessed. RESULTS: The most constant measurement with the least change on the repeated radiographs was that for pelvic morphology (PR-S1 angle) followed by length of the pelvic radius, pelvic alignment over the hips (pelvic angle), and total lumbopelvic (PR-T12) and lumbosacral (T12-S1) lordosis. Other longitudinal measurements, including those for thoracic kyphosis and spinal balance by a plumbline, showed greater change. Measurements for pelvic morphology by the pelvic radius technique were correlative with standing total lumbosacral lordosis, regional lumbopelvic lordosis, pelvic alignment, pelvic radius length, and gender (P< or = 0.006 for each). The correlations between total and regional lumbopelvic lordosis and pelvic alignment measurements were even higher(P<0.0001). Of possible clinical importance was the finding that standard measurements for lordosis were dependent on individual pelvic morphology quantitated by the pelvic radius technique. CONCLUSIONS: In all of the sagittally balanced subjects studied, "congruent" spinopelvic alignment on all 105 standing lateral radiographs could be defined by four parameters using the pelvic radius technique: total lumbopelvic lordosis (PR T12), incorporating complementary angles for lumbosacral lordosis (T12-S1), and pelvic morphology (PR-S1 angle) that summarily were always between -69 degrees to -116 degrees (+/-3 degrees ); centered pelvic alignment over the hips, as determined by the pelvic angle, that was always between -3 degrees to -32 degrees (+/-2 degrees ); compensated spinal balance, with a sagittal plumbline from the center of the T4 body always posterior to the hip axis as well as the center of the L4 vertebral body; and a concordant T4-T12 kyphosis/PR-T12 lordosis ratio that was always negative and between 0.15 to 0.75. [Key words: congruent alignment, pelvic radius technique, pelvic morphology, lumbopelvic lordosis, lumbosacral lordosis] PMID- 11064529 TI - Point of view PMID- 11064528 TI - Assessing the needs of patients in pain: a matter of opinion? AB - STUDY DESIGN: A prospective cohort study including patients with nonspecific spinal pain was performed. OBJECTIVES: To investigate whether the use of expert judgment in routine practice can provide a basis for reliable decision making concerning the need for intervention in patients with spinal pain and their ability to benefit from treatment. SUMMARY OF BACKGROUND DATA: A wide range of instruments and techniques are used to assess and treat patients with spinal pain. Many instruments are used without being clinimetrically tested. METHODS: A questionnaire concerning the patients' need of treatment and their potential to assimilate it was sent to experts in the health care arena: physicians, physical therapists, social insurance officers. The experts included were those connected with patients participating in a larger outcome study. Two cohorts of patients (sample 1, n = 217; sample 2, n = 257) were followed for 6 and 12 months, during which time the patients' health and work status were mapped. RESULTS: No acceptable agreement was found between any of the experts' ratings of patients' needs and potential for rehabilitation. Logistic regression showed that the experts' judgments were based almost solely on the age of the patient. The prediction analyses showed that the most consistent predictor of the patients' status at the 6-month follow-up assessment was the patients' own belief in the existence of effective treatments and their perceived ability for learning to cope with the condition. CONCLUSIONS: Expert judgment as exercised in routine practice cannot be used as basis for reliable decision making concerning the need of the patient with spinal pain for intervention and the patient's ability to benefit from treatment. PMID- 11064530 TI - Can chronic disability be prevented? A randomized trial of a cognitive-behavior intervention and two forms of information for patients with spinal pain. AB - STUDY DESIGN: A randomized controlled design superimposed on treatment as usual was used to compare the effects of a cognitive-behavior intervention aimed at preventing chronicity with two different forms of information. OBJECTIVE: To develop a coping-oriented preventive intervention applicable in primary care, and to compare its impact with educational information. SUMMARY OF BACKGROUND DATA: Preventing long-term disability resulting from spinal pain has proved difficult. The information provided by health care professions and early interventions aimed at preventing long-term disability may be important, but little scientific evidence exists concerning their use. METHODS: A protocol for a six-session cognitive-behavior group intervention was developed on the basis of earlier research. The main focus was to prevent long-term disability by changing patients' behaviors and beliefs so they can cope better with their problems. Comparison groups received either a pamphlet shown earlier to have an effect, or a more extensive information package consisting of six installments. All the groups continued to receive treatment as usual in primary care. There were 243 patients with acute or subacute spinal pain who perceived that they were at risk for developing a chronic problem. These patients were randomized to the cognitive behavioral intervention or one of the two information groups. Because the aim was to prevent long-term disability, the key outcome variables at the 1-year follow up assessment were sick absenteeism and health care use. Other variables were pain, function, fear-avoidance beliefs, and cognitions. RESULTS: The comparison groups reported benefits. However, the risk for a long-term sick absence developing was lowered ninefold for the cognitive-behavior intervention group as compared with the risk for the information groups (relative risk, 9.3). Participants in the cognitive-behavior group also reported a significant decrease in perceived risk. In addition, the cognitive-behavior group demonstrated a significant decrease in physician and physical therapy use as compared with two groups receiving information, in which such use increased. All three groups tended to improve on the variables of pain, fear-avoidance, and cognitions. CONCLUSIONS: This study demonstrates that a cognitive-behavior group intervention can lower the risk of a long-term disability developing. These findings underscore the significance of early interventions that specifically aim to prevent chronic problems. This approach might be applied to primary care settings. PMID- 11064531 TI - Posterior spinal shortening for paraplegia after vertebral collapse caused by osteoporosis. AB - STUDY DESIGN: Case report of a patient who underwent a new surgical procedure for paraplegia after vertebral collapse due to osteoporosis. OBJECTIVES: To propose a new approach to posterior spinal fusion surgery for osteoporotic patients. SUMMARY OF BACKGROUND DATA: Surgical treatment was performed on a paraplegic patient after vertebral collapse due to osteoporosis. However, the surgery was difficult because implants such as hooks and screws often dislodged during the treatment. The poor holding power of these implants to the osteoporotic spine is a challenging problem in this treatment. METHODS: When a fractured vertebra is shortened by resecting the posterior part of the spine and the application of a compression force, a short vertebra is produced. As a result, the thoracic kyphosis decreases and the force pushing the upper thoracic spine inferio ventrally also decreases. RESULTS: A 74-year-old woman with T12 vertebral collapse was treated with this new method. Lateral Cobb angle (T10-L2) was reduced from 26 to 4 degrees after surgery. The shortened vertebral body united, and after 33 months, the implant had not dislodged and no loss of correction was seen. CONCLUSION: The posterior spinal shortening can be a choice for treating delayed paraplegia after osteoporotic vertebral fracture. PMID- 11064532 TI - Horner's syndrome after posterior spinal fusion in a child: a case report. AB - STUDY DESIGN: A case report of a complication after posterior spinal fusion. OBJECTIVES: To present the clinical findings of a Horner's syndrome after posterior spinal fusion. SUMMARY OF BACKGROUND DATA: A 14-year-old girl underwent posterior spinal fusion for idiopathic scoliosis. METHODS: Clinical examination and pharmacologic pupillary testing were used to diagnose Horner's syndrome. RESULTS: After surgery, the patient developed a left-sided Horner's syndrome. The Horner's syndrome had resolved 6 months after surgery except for slight ptosis. CONCLUSIONS: This is the first reported case of Horner's syndrome occurring after posterior spinal fusion without the use of epidural analgesia. PMID- 11064533 TI - Historical perspective: history of spinal surgery. AB - The surgical treatment of spinal disorders did not develop before the 1970s of the last century. Previously limited technical possibilities and the danger of infections spinal surgery could not spread wider. This article reviews the history of spinal surgery from first trials as mentioned in the papyrus Smith in 1550 B.C. in Egypt to advanced techniques of today. PMID- 11064534 TI - Re: Bridging the gap between science and practice in managing low back pain: a comprehensive care system in a health maintenance organization setting. PMID- 11064535 TI - Re: the association between static pelvic asymmetry and low back pain. PMID- 11064536 TI - The use of revised Oswestry Disability Questionnaire. PMID- 11064538 TI - Imagery PMID- 11064537 TI - Re: post-traumatic findings of the spine after earlier vertebral fracture in young patients. PMID- 11064539 TI - Coronary artery bypass grafting in the presence of atheromatous or calcified aorta: On-pump or off-pump? PMID- 11064540 TI - The TRUCAB, the "H" graft, and the steal syndrome. PMID- 11064541 TI - Posterior thoracotomy for reoperative coronary artery bypass grafting without cardiopulmonary bypass: perioperative results. AB - BACKGROUND: This retrospective study evaluates morbidity and mortality of reoperative coronary artery bypass grafting (CABG) without cardiopulmonary bypass (CPB) using a posterior thoracotomy to revascularize the lateral aspect of the heart. METHODS: From January 1995 to July 1999, reoperative CABG without CPB was performed on 67 selected patients using a left posterior thoracotomy approach. Preoperative risk factors, postoperative mortality, and major complications were derived from the New York State database. RESULTS: All patients were operated on without CPB. A total of 1.3 grafts per patient were performed. Freedom from major complications was 95.5%. There were no postoperative cerebro-vascular accidents (CVA) or new neurological deficits. Two patients (3%) had a perioperative acute myocardial infarction. The actual mortality rate was 4.5% (3/67), the expected mortality was 5.1% and the calculated risk adjusted mortality was 2.1%. CONCLUSIONS: Reoperative CABG without CPB to revascularize selected coronary artery targets can be safely performed using a posterior thoracotomy approach. PMID- 11064542 TI - Transaortic repair of mitral regurgitation. AB - BACKGROUND: In the operative management of mitral regurgitation (MR) associated with aortic valve disease, a transaortic approach combining the bowtie mitral valve repair with replacement of the aortic valve appears to offer a less invasive and technically simple, expeditious alternative to conventional left atriotomy and Carpentier style repair. METHODS: Between February 1997 and December 1999, four patients underwent a bowtie repair of the mitral valve via the aortic root with concomitant aortic valve replacement. The diagnosis of MR was established and followed postoperatively by echocardiogram. The operative technique involved a transaortic annular approach to the mitral valve with a single edge-to-edge suture approximating the prolapsing posterior mitral leaflet to a normal segment of the anterior leaflet. RESULTS: There were no operative mortalities. Mean cross-clamp time for both valve procedures was 104 +/- 24 min and cardiopulmonary bypass was 155 +/- 31. Mean postoperative cardiac output was 5 +/- 1 L/min. Semiquantitative estimation of mitral regurgitation by doppler improved from a mean of 3.2 +/- 0.5 preoperatively to a mean of 0.25 +/- 0.5 (p = 0.0052) postoperatively, while ejection fraction (EF) remained stable (48 +/- 9% preoperatively and 49 +/- 9% prior to discharge). One patient with rheumatic mitral pathology had a mild increased mitral gradient which did not resolve with takedown of the bowtie repair. Mitral stenosis was not evident in any of the other patients. CONCLUSIONS: Our initial experience with the combined transaortic bowtie repair and aortic valve replacement has demonstrated that this approach is very quick, feasible, effective, and technically simple with gratifying midterm results. PMID- 11064543 TI - Total endoscopic off-pump coronary artery bypass grafting. AB - Two cases of totally endoscopic off-pump coronary artery bypass grafting (TECAB) of the left internal thoracic artery to the left anterior descending artery using the da Vincitrade mark telemanipulation system (Intuitive Surgical, Mountain View, CA) are described. A new articulating endoscopic stabilizer with cleats was developed to enable endoscopic anchoring of silastic vessel loops for vascular occlusion. Newly created attachments for irrigation and suction, along with active robotic enhanced assistance by a second surgical console, permitted our group to perform for the first time a truly endoscopic bypass grafting without any thoracotomy. PMID- 11064544 TI - Functional and morphologic assessment of saphenous veins harvested with minimally invasive techniques using a modified laryngoscope. AB - BACKGROUND: Minimally invasive saphenous vein harvesting techniques have been shown to reduce postoperative morbidity. Commercially available and often disposable instruments add significant costs to the operation. To lower expenses and to reduce postoperative morbidity, we used an ordinary laryngoscope fitted with a modified # 3 Heine blade for harvesting the greater saphenous vein for coronary artery bypass surgery. OBJECTIVE: To assess the integrity and function of the autologous, undistended, long saphenous vein harvested by a modified laryngoscope. METHODS: Morphology was examined by light and scanning electron microscopy. Endothelial function was assessed by vascular reactivity in an isolated organ bath. Veins, randomly taken and prepared traditionally, served as a control group. Contractile function was measured in response to potassium chloride. Endothelium-dependent relaxation was assessed by use of acetylcholine and calculated as percentage relaxation. RESULTS: There were no significant differences, in response to the constricting or dilating agent, in vein rings taken with the modified laryngoscope compared with the traditional 'open' technique (n = 10, p > 0.05 by ANOVA). Histologic examination by light and scanning electron microscopy showed no significant damage to the endothelial layer. CONCLUSION: Minimally invasive saphenous vein harvesting, using a modified laryngoscope yields morphologically and biologically intact veins. PMID- 11064545 TI - Development of a prosthetic coronary artery bypass graft. AB - BACKGROUND: The patient population undergoing repeat coronary revascularization is increasing, with estimates of approximately 15% of these patients in need of alternative conduits. Pre-existing conditions may limit the availability of suitable autogenous vessels for complete coronary revascularization. As an alternative, previous investigators have attempted to develop prosthetic conduits for coronary artery bypass grafting (CABG), but as yet without clinical success. To be of clinical benefit, a prosthetic graft would require demonstrated patency at least as good as a marginal quality autogenous vessel used in the same position, without unexpected adverse effects. The use of a prosthetic graft may also reduce surgical complications associated with conduit harvesting and thereby enhance the speed of patient recovery. METHODS: Our group has investigated the potential of a novel synthetic small diameter vascular graft (2.5- to 3.5-mm diameter), as an alternative conduit for CABG. The graft is designed with three distinct layers composed of Thoralon, a proprietary polyetherurethaneurea with a silicone-based surface modifying additive. This biomaterial is the same material successfully used for the thromboresistant blood-contacting surfaces of an FDA approved and clinically successful ventricular assist device. RESULTS: Aria grafts underwent extensive preclinical testing in sheep with results to over one year that demonstrate the graft's biocompatibility, biodurability, and ability to maintain patency in both peripheral access graft and coronary applications. Compassionate use human coronary implants have been performed in 27 patients in the coronary position in Canada and Europe. Although incomplete, the data demonstrate no device related serious injury or and all surviving patients have remained symptom free. CONCLUSIONS: A prosthetic coronary artery bypass graft has been developed and has undergone extensive pre-clinical testing and preliminary clinical use. Based upon the results, a prospective, randomized, controlled clinical study, the AEGIS/Canada (AlternativE Graft Investigational Study) trial using the Aria graft in the coronary position in human patients is underway. Additionally, a pre-IDE submission has been submitted to the FDA to expand the AEGIS clinical trial device to the United States. PMID- 11064546 TI - Minimally invasive coronary revascularization in women: A safe approach for a high-risk group. AB - PURPOSE: Female gender has been shown to be an independent risk factor for mortality in coronary artery bypass graft (CABG) surgery. This report analyzes our early outcomes in 304 women who underwent off-pump coronary artery bypass (OPCAB) surgery at the Washington Hospital Center (Washington, DC) over the last 3 years to determine whether this is a safe approach for coronary bypass in women. METHODS: A retrospective review of 5528 cases of CABG bypass (on-pump) and 840 cases of OPCAB surgery, from June 1996 to July 1999, was performed. Women accounted for 1527 (27.6%) of the on-pump bypass cases and 304 (36.2%) of the OPCABs. All cases without cardiopulmonary bypass were included, with the majority of the most recent cases being multivessel revascularization. The data for analysis were obtained from our cardiac surgery database and included cases from all surgeons operating at the Washington Hospital Center, although the majority of off-pump cases were performed by only a few of these surgeons. RESULTS: The two groups were similar with respect to urgent cases, redos, and other comorbities including preoperative congestive heart failure, peripheral vascular disease, transient ischemic attack (TIA), cerebral vascular accident, and previous myocardial infarction. The mean age for the two groups was similar, 67 years for the off-pump group and 66 years for the on-pump group. The absolute number of all off-pump cases increased each year (from 175 to a total of 373), representing a corresponding increase in percentage of all coronary artery bypass procedures (from 9% to 16%). Of the total number of patients undergoing CABG, the percentage of women who underwent OPCAB doubled from 3% to 6% over the time period analyzed. The percentage of single-vessel cases in the off-pump group fell from 88% to 41% as multivessel bypasses became more routine However, the percentage of patients aged > 75 years was greater for the off-pump group (30%) than for the on-pump group (24%). Otherwise, the two groups differed only in diabetic disease (36% off-pump compared with 46% on-pump; p = 0.001) and previous transcatheter therapy (38% off-pump compared with 29% on-pump; p = 0.003). Patients who had OPCABs received fewer postoperative transfusions (40%) than the on-pump group (59%; p < 0.001). The off-pump group also had fewer neurological complications in the form of TIAs or strokes (0.3%) compared with the on-pump group (3.5%; p = 0.001). The mortality rate was 2.3% off -pump versus 4.1% on pump but did not reach statistical significance in this study (p =.12). CONCLUSION: Myocardial revascularization in women can be performed safely without cardiopulmonary bypass. In our series, the mortality for women receiving off-pump revascularization was lower than the on-pump cohorts despite an older age and higher incidence of diabetes. Although the absolute mortality rates did not reach statistical significance, we were encouraged that the mortality rate for women operated on without CPB dropped to the mortality rate typically seen in men. We also observed a favorable tendency in the off-pump group for a shorter length of stay and a lower incidences of transient ischemic attacks, strokes, post-op bleeding, and blood transfusions. A larger series of patients with multivariate analysis and/or a prospective trial will need to be analyzed in order to confirm our findings. PMID- 11064547 TI - Ventriculocoronary artery bypass (VCAB), a novel approach to myocardial revascularization. AB - BACKGROUND: The long-term patency rate of saphenous vein grafts for myocardial revascularization is poor (50% at 10 years). Half of the patent grafts develop severe atherosclerosis. In this paper, we report on an implantation technique and an in vivo evaluation of a device that creates a ventriculocoronary artery bypass (VCAB), a permanent transmyocardial channel between the left ventricle and a coronary artery. METHODS: An L-shaped titanium tube with an exterior polyester cuff was implanted from the base of the left ventricle to the proximal left anterior descending coronary artery in 11 juvenile domestic pigs using a beating heart approach. Flow rates were measured at implantation. Patency was assessed when explanted at 2 weeks. RESULTS: The flow rate through the device after implantation was 76% of baseline. Forward flow occurred during systole. The patency rate was 91% at 2 weeks. Histologic analysis showed the formation of an organizing tissue at the coronary interface. CONCLUSIONS: These preliminary studies show the promise of perfusing ischemic myocardium with systolic flow. Patency of the transmyocardial titanium conduit was excellent at 2 weeks and warrants longer duration studies. PMID- 11064548 TI - The use of a novel tissue sealant as a hemostatic adjunct in cardiac surgery. AB - BACKGROUND: In spite of advances in the management of bleeding associated with cardiac surgery, hemorrhage remains a troublesome problem, particularly in complex cases and high risk patients. In minimally invasive cardiac surgery, limited exposure and tight quarters may make accurate suturing difficult, and increase the risk of surgical bleeding. A surgical sealant that effectively prevents suture line bleeding would be a valuable resource for cardiac surgeons and might help to facilitate minimal access cases. METHODS: We undertook acute canine studies with a new polyethylene glycol-based tissue sealant (FocalSeal, Focal, Inc., Lexington, MA) to determine its effectiveness in controlling bleeding from graduated needle punctures sites in the arteries of heparinized animals. For chronic canine studies, the sealant was applied to the suture line of a left internal mammary artery (LIMA) to left anterior descending (LAD) anastomoses. The anastomoses were then evaluated for patency and tissue reaction after a three-month recovery period. RESULTS: The sealant prevented bleeding from arterial puncture wounds up to 2.5 mm in diameter. Three months following the application of sealant to coronary anastomoses, no adverse tissue reaction was found on histologic examination. All anastomoses treated with the sealant remained patent. CONCLUSION: When applied as a hemostatic adjunct to sutures at a coronary anastomosis, the sealant appears to be an effective means of preventing bleeding without adverse tissue reaction or scarring. PMID- 11064549 TI - ECG of the month. Concordance or discordance. Complete left bundle branch block. PMID- 11064550 TI - Radiology case of the month. Painful eye. Orbital myositis (orbital pseodotumor). PMID- 11064551 TI - Journal 100 & 150 years ago. Some interesting notes. PMID- 11064552 TI - Long-term sterilization failure: twenty-three years. AB - This case presents the longest time interval from tubal sterilization to failure by ectopic pregnancy of which we or our colleagues have ever heard. This multipara had a postpartum sterilization procedure performed at one University Hospital; 23 years later she was admitted to another University Hospital with a hemoperitoneum due to a ruptured ectopic pregnancy. Verification was affirmed by examination of the records of both hospitals. PMID- 11064553 TI - Mediation in medical liability litigation. PMID- 11064554 TI - The Internet for Louisiana physicians. AB - Fewer than 50% of Louisiana physicians actively use the Internet, and many of them confine their usage to e-mailing among family and friends. The purpose of this article is to acquaint the reader with many of the benefits of exploiting the incredible potential of this technological invention. I provide addresses and information about sites that I believe warrant usage by our colleagues. Of the vast smorgasbord of data available we highlight educational Web sites for professionals and the public, how to determine credibility of information, clinical research of scientific articles, computer security, federal and state government sites, newspapers, political and socioeconomic functions, medical supply shops, e-mail and other computerized communication, electronic medical records, personal or professional Web sites, and future medical internet uses. It is hoped that this process will encourage nonparticipating colleagues to begin using this modality while also supplying sites that current users may not yet have discovered. PMID- 11064555 TI - Providing access to prescription drugs for America's retirees. PMID- 11064556 TI - Government affairs at MAG. PMID- 11064557 TI - Private sector advocacy provides tremendous benefit to members. PMID- 11064558 TI - Extra! extra! Read all about it: the all new MAG Web site. PMID- 11064559 TI - E-Lobbying: legislative grassroots Web page. PMID- 11064560 TI - Expanding access to insurance coverage for the uninsured. PMID- 11064561 TI - Department of Education: MAG University, thought leaders, and more. PMID- 11064562 TI - William Augustus Newell: physician, congressman, governor. PMID- 11064563 TI - Heart transplantation services overdue for southern New Jersey. PMID- 11064564 TI - Introduction to clinical positron emission tomography. AB - Positron emission tomography (PET) scanning with [18F] fluorodeoxyglucose (FDG) has developed over 25 years into a valuable clinical tool used mainly for the evaluation of oncology patients. The purpose of this article is to provide an introduction to the basic scientific concepts of FDG-PET imaging as well as to illustrate its clinical utility. PMID- 11064565 TI - HIV resistance testing: a new clinical tool. AB - Drug resistance threatens to erase the recent gains made in treating HIV infection. HIV resistance testing offers clinicians the ability to screen patients for resistant strains and adjust treatment accordingly. Resistance testing is recommended for patients on antiretroviral agents with virologic failure or with suboptimal suppression of viral load after initiation of antiretroviral therapy. Because of the potential transmission of resistant strains, testing should be considered for patients with acute infection. Although the care of many AIDS patients is complex and should be managed by a physician with experience and expertise in HIV disease, all practitioners should be familiar with the types of testing available and their limitations. PMID- 11064566 TI - The sharing network: the first decade. PMID- 11064567 TI - Negotiating effective managed care contracts. PMID- 11064568 TI - [Results of limb-saving surgery and prognostic factors in patients with osteosarcoma]. AB - Ninety six patients with high-grade osteosarcoma of the extremities were treated between 1986 and 1997 in the authors institution. They were divided into three groups: in group I, all of 75 patients with non-metastatic OS received intensive chemotherapy and underwent surgery. In group II, 9 patients already had metastases at the time of referral. In group III, 12 patients received chemotherapy in delayed or suboptimal form. In group I, local recurrences occurred in 7 per cent (3 patients), metastases in 20 per cent of the patients with limb-saving, whereas these numbers were 3 per cent and 38 per cent in the amputation group. The 5-year disease free survival (DFS) was 72% v 69% in the limb-saving and amputation group, respectively. In groups II and III, 5-year DFS was extremely poor, 10 and 20% only. With univariate analysis, factors having a positive influence on the survival were: tumor volume < 60 cm3, wide or radical surgical margin, distal location of osteosarcoma, cartilagineous ground substance less than 20% and response to chemotherapy. The last 4 variables maintained their significance in the multivariate Cox model as well. Age > 30 showed indirect negative influence on the final outcome (enhanced intolerability to the drugs and less co-operability of the patients etc.). This data confirm the competence of the limb-saving surgery at certain indications beside the amputation. PMID- 11064569 TI - [Coronary revascularization on beating heart]. AB - At authors clinic 563 coronary artery bypass graft procedures were done from 1. January 1998 to 15. September 1999. 134 coronary revascularizations were performed from the same surgical team on beating heart using the same surgical technic at normothermia. Out of them, 98 operations were done with and 36 without ECC support respectively. Patients were operated on with this sophisticated technic exclusively during this period. Conversion to cardiac arrest was not necessary in any case. Male/female ratio was 76/58, 84% of the patients suffered from one or more preoperative myocardial infarction. The average bypass number was 2.7/patient. Arterial revascularization was performed in 94%. Postoperative mortality was 2.2% (3 pts). The postoperative inotropic demand, myocardial infarction rate (CK-MB release) and the amount of bleeding was very low. Mean postoperative stay in the ICU decreased to the average of 19 hrs. Patients normally left hospital on the 7th postoperative day. This operative technique- performed by an experienced surgeon--is less invasive for the heart and other organ systems, the incidence of complications can be decreased and the principle of complete revascularization is not compromised. PMID- 11064570 TI - [Prognostic value of cell heterogeneity in cervical cancer determined by digital image analyzer of DNA content]. AB - Frequency and prognostic value of cell heterogeneity in FIGO 1a-2a cervical cancer was examined, in 66 of patients underwent Wertheim type hysterectomy between 1989 and 1995 in National Institute of Cancer, Budapest, Hungary. A newly developed DNA image analyses (DNACE) was used in paraffin embedded tissues after enzymatic hydrolyses for evaluation of the DNA content in cervical cancer. In 30.3% of examined tissues (20/66) two subgroups was found. There was significant differences in the DNA indexes (DI) between the subgroups (p = 0.0001). In the remaining 69.7% of the cases only one subgroup was present. The frequency of two subgroups was higher between aneuploid (78.4%), or hyperploid (81.5%) type cervical cancer, however there was no significant difference between the two groups. On the other hand there was significant difference in the presence of two subgroups between the well and less differentiated cervical cancer. The frequency was higher between the less differentiated groups (p = 0.02). Looking at the prognostic value of subgroups, there was no significant correlation between the heterogeneity of cervical cancer and FIGO stage, or lymph node metastasis (p = 0.6855), or vascular/lymphatic space infiltration (p = 0.2558), or invasiveness of cancer (0.0823). There was neither significant value found between the outcome of disease and the number of subgroups present (p = 0.8738). It is though that the present of cellular heterogeneity in cervical cancer is connected with the differentiation of the cancer cells, and can be a good prognostic value in the anticipation of the aggressiveness of cervical cancer. Looking at the present result, there was no significant connection between the heterogeneity of cervical cancer and the outcome of the disease, so further examination should be done. PMID- 11064571 TI - [Current issues in gastroeneterologic rehabilitation]. AB - In Hungary gastroenteric diseases' rehabilitation, which is one of the youngest branch of medical rehabilitation, has grown out of traditional, sanatorial care from the 70s. Its main feature that it is based on a biopsychosocial attitude towards patients as a team-work. Though it comprises traditional, medical diagnosis and treatment, medicine and medical technology are not the primary resources of gastroenteric diseases rehabilitation: it is mainly built on a friendly and co-operating relations with the patients, which helps doctors to exert their mental and intellectual influence promoting their patients recovery. Therefore, gastroenteric diseases' rehabilitation can be sharply separated from active treatment and care. The article gives full details of the possible methods of gastroenteric diseases' rehabilitation, such as diagnosis, medicinal treatment, psychotherapy, dietotherapy, physiotherapy and regimen guidance. In addition, it is emphasized that under sanatorial circumstances the effects of rehabilitation, especially of regimen guidance multiplies. Furthermore it gives numerical data about the present situation of gastroenteric diseases' rehabilitation in Hungary. Considering the data gained from the special rehabilitation hospital located in Visegrad, the article touches upon the illnesses calling for this kind of rehabilitative treatment not passing over the fact of multimorbidity, especially important in rehabilitation. PMID- 11064572 TI - [Belated commemoration of Mrs. Dr. Balo, Dr. Ilona Banga]. PMID- 11064573 TI - [Hungarian Medical Archive--100 years ago]. PMID- 11064574 TI - [In memoriam veritatis: Ophthalmology Department of the Buda Mercy Hospital, later National Institute of Rheumatology and Physiotherapy until 1978]. PMID- 11064575 TI - [Injuries caused by snake bites]. PMID- 11064576 TI - [The pampiniform plexus in the chronic phase of human Chagas disease: histologic assessment]. AB - The occurrence of Trypanosoma cruzi intracellular clusters and phlebitis was searched for on pampiniform plexus vein walls of chronic chagasic patients. For this purpose, 23 pairs of spermatic cords, epididymides and testes (17 from chagasic patients and 6 from non-chagasic controls) were obtained, at autopsy. Trypanosoma cruzi was investigated by immuno-histochemistry on slides obtained from several sections of the gonads and vessels of each case. Only discrete and focal undetermined chronic phlebitis was observed, with no parasites, in 5 chagasics (bilateral in 3) and 2 controls (chi 2: p < 0.10), and discrete mononuclear interstitial infiltration in the funiculi of 13 chagasics and 5 controls (chi 2: p < 0.75). In conclusion, on the contrary to that published regarding the supra-renal central veins, it seems that the hormonal environment provided by testosterone does not favor the infection of the gonadal vessel wall. PMID- 11064577 TI - [Epidemiology and ecology of dermatophytoses in the City of Fortaleza: Trichophyton tonsurans as important emerging pathogen of Tinea capitis]. AB - Dermatophytosis is the most common skin infectious disturbance in the world. In this research 2,297 patients were evaluated with suspected clinical lesions of dermatophytosis. It was observed that, 534 (23.2%) patients tested positive for dermatophytes. T. rubrum was the most prevalent specie (49.6%; p < or = 0.05), followed by T. tonsurans (34.4%), M. canis (7%) and T. mentagrophytes (6.2%). When the species isolated was correlated with the respective anatomical localization, it was observed that T. tonsurans was the most frequent isolated in scalp lesions (73.9%; p < or = 0.01). On the other hand, T. rubrum was the main specie involved in body lesions (72.8%; p < or = 0.05). Therefore, in scalp infections it was observed that, there was an absolute prevalence of T. tonsurans. This evidence is different from the statistical data collected in the southeast and south of Brazil, as well as from other areas of the world, which still show M. canis as the most frequent microorganism isolated in Tinea capitis. PMID- 11064578 TI - Absence of both circadian rhythm and Trypanosoma cruzi periodicity with xenodiagnosis in chronic chagasic individuals. AB - Xenodiagnoses were performed every 3 hours using 10 Triatoma infestans 3rd instar for 24 to 72 hours, in 18 chronic chagasics with positive serology and/or xenodiagnosis. There was no statistically significant difference in the positivity of assays performed during the day (9:00 to 18:00 h) compared to those performed at night (21:00 to 6:00 h), (chi 2 = 0.1526 p = 0.696). Xenodiagnosis was performed in ten of the patients for 13 successive days but there was no periodicity detected in the positive assays. PMID- 11064579 TI - [Assessment of schistosomiasis and other intestinal parasitoses in school children of the Bambui municipality, Minas Gerais, Brazil]. AB - This work was carried out with the purpose of determining the prevalence of intestinal parasitoses in the school children of Bambui, through parasitological examinations (direct and Kato-Katz methods) and reevaluating the snails' breeding places described in the county. Of the 2,091 school children examined, 20.1% had at least one parasitic infection. Giardia lamblia, Entamoeba coli, Ascaris lumbricoides and hookworm are the most frequent parasites, with a prevalence of 6.2%, 6.2%, 4.8% and 1.4%, respectively. The hookworms were significantly more frequent among students from the rural area and in those aged over 14 years, while the prevalence of E. coli was greater in the urban area and the G. lamblia was more frequent in children under 6 years old. Only three children shed eggs of Schistosoma mansoni. The only intermediate host found was Biomphalaria glabrata and none of them was shedding cercariae of S. mansoni. When these data were compared with data from other surveys previously effected in the county, a decline was observed in the prevalence of all parasites. Some hypotheses which may explain this decline are discussed, such as: intense urbanization process and improvement of social-sanitary conditions of the county. PMID- 11064580 TI - [Enzyme typing of species of the genus Candida isolated from the oral cavity]. AB - The production of phospholipase and proteinase exoenzymes was evaluated in seventy nine samples of Candida isolated from the oral cavity of patients with oral lesions characteristic of candidosis and from individuals presenting a clinically normal mouth, attended at the University of Dentistry of Ribeirao Preto USP. Among the strains of C. albicans isolated from oral lesions, the phospholipase and proteinase were detected in 83.3% and 66.7%, respectively. C. tropicalis and C. parapsilosis produced only proteinase. Regarding the isolated strains from niches without lesions, out of a total of 32 C. albicans, 71.9% presented phospholipase and 68.7% proteinase. C. tropicalis only presented the enzyme proteinase, C. glabrata, C. krusei, C. guilliermondii and Candida spp did not present any of the exoenzymes. Among the samples of C. albicans from both groups, the enzymotype 22 (positive phospholipase and proteinase weakly positive), was prevalent. Different enzymotypes of the same species were detected in samples collected from the same patient. PMID- 11064581 TI - [Anopheles species in the municipality of Pinheiro (Maranhao), endemic area of malaria]. AB - The Anophelae species of the municipal district of Pinheiro, Maranhao State, Brazil, were studied, considering their variety, relative abundance, seasonal fluctuation, preference by the peri and intra domiciles as well as the hours of hematophagism. The females were captured using human bait, on a monthly basis from May/1998 to April/1999, from 6 p.m. to 6 a.m. A total of 1,321 specimens of 10 species were captured, all belonging to the subgenus Nyssorhynchus. The most frequent species were A. argyritarsis (62%), A. darlingi (21.7%), A. albitarsis (4.5%), A. galvaoi (4.0%), A. triannulatus (3.1%) and A. evansae (2.8%). The species A. nuneztovari, A. braziliensis, A. rondoni and A. strodei together represented 1.9%. The anophelines occurred all year round, with no significant difference in the number of specimens captured between the rainy (51.7%) and dry season (48.3%). The frequency of the females in the peridomicile was significantly higher (p < 0.01) (82%), than in the intradomicile (18%), preferring to suck blood at dusk and in the first hours of the night. PMID- 11064582 TI - [Gonorrhea]. AB - Gonorrhea is a common bacterial infection caused by Neisseria gonorrhoeae, a Gram negative diplococcus that is transmitted almost exclusively by sexual contact or perinatally. It primarily affects the mucous membranes of the lower genital tract and less frequently those of the rectum, oropharynx, and conjunctivae. Ascending genital infection in women leads to the predominant complication, acute salpingitis, one of the most common causes of female infertility in the world. Since the 1990s, a remarkable surge of information ensued regarding the pathogenesis of gonorrhea and its agent. Gonorrhea has proven difficult to control in most populations and remains a prime example of the influence that social, behavioral, and demographic factors can have on the epidemiology of an infectious disease. The management of gonorrhea and other sexually transmitted infections requires both treatment of the patient as an individual and of his or her sexual partner(s) as a public health measure to interrupt the onward spread of infection and prevent long-term complications. PMID- 11064583 TI - [Dengue: a new approach]. AB - Previous definition of classic dengue, with or without bleeding, and of dengue hemorrhagic fever (DHF) that may evolve without bleeding and with or without dengue shock syndrome (DSS) are reviewed here. The classical approach to the diagnosis and treatment of dengue, although useful in the past, nowadays breeds confusion and adds a burden to the physician's task of decision-making regarding the treatment of patients with severe forms of the disease. The classification of dengue proposed in this paper, and summarized in a diagram, incorporates new concepts about sepsis, systemic inflammatory response syndrome (SIRS), and acute respiratory distress syndrome (ARDS). This new approach, in our view, is a useful guide to initial evaluation and treatment of the disease. It also approximates the dengue syndrome to other protocols and medical procedures routinely used in intensive care units, making it easier to be followed by the health personnel working in areas subject to epidemic bursts. PMID- 11064584 TI - Aplastic crisis caused by parvovirus B19 in an adult patient with sickle-cell disease. AB - We describe a case of aplastic crisis caused by parvovirus B19 in an adult sickle cell patient presenting with paleness, tiredness, fainting and dyspnea. The absence of reticulocytes lead to the diagnosis. Anti-B19 IgM and IgG were detected. Reticulocytopenia in patients with hereditary hemolytic anemia suggests B19 infection. PMID- 11064585 TI - Pulmonary adiaspiromycosis: report of two cases. AB - Two cases of human pulmonary adiaspiromycosis are reported. Patients were 29 and 54-year-old males, farm workers, with "grippe-like" symptoms and radiographic findings suggestive of granulomatous interstitial disease. Transthoracoscopic and transbronchial biopsies were performed. Pulmonary function was measured by spirometry. One patient used ketoconazole. Possibility of misdiagnosed pulmonary adiaspiromycosis is emphasized. PMID- 11064586 TI - How prevalent is Plasmodium malariae in Rondonia, western Brazilian Amazon? AB - We have compared results of Plasmodium species identification obtained with conventional on-site microscopy of Giemsa-stained thick smears (GTS) and a semi nested polymerase chain reaction (PCR) in 96 malaria patients from Rondonia, Western Brazilian Amazon. Mixed-species infections were detected by PCR in 30% patients, but no such case had been found on GTS. Moreover, P. malariae infections were detected in 9 of 96 patients (10%) by PCR, but were not identified by local microscopists. The potential impact of species misidentification on malaria treatment and control is discussed. PMID- 11064587 TI - [Contribution to the morphologic diagnosis of lung adiaspiromycosis]. AB - The diagnosis of adiaspiromycosis is usually based on lung sections stained by hematoxylin-eosin, periodic acid Schiff and methenamine silver. Authors describe the fungus aspect examined by mucicarmin, picro-sirius and Congo red methods, including polarized light microscopy. In doubtful cases, these methods could contribute to histopathological diagnosis of Emmonsia parva var crescens. PMID- 11064588 TI - [Influenza, yes; grippe, no]. PMID- 11064589 TI - [Strategy for diagnosing hematological malignancies--a proposal from hematologists to laboratory medicine]. AB - Many laboratory methods are used to diagnose acute myelocytic leukemia(AML) including morphological analysis, phenotypic analysis, chromosomal and genetic analysis of leukemic blasts. The FAB classification is widely used for the diagnosis and classification of AML because of its convenience. This classification is mainly based on morphological and cytochemical results which are well established and economical methods. Phenotypic analyses using flow cytometry provides useful information regarding the lineage and maturation status of leukemic cells. Chromosomal and genetic data are thought to reflect the biological characteristics of leukemic cells and certain chromosomal abnormalities strongly associated with the good or poor prognoses of AML patients. Genetic analysis of AML blasts became available recently in a clinical field, along with progress in molecular analysis of leukemia but its clinical significance in AML has not been well established yet. When clinicians see patients, the diagnosis of AML is usually made based on the morphological and phenotypic data of peripheral and bone marrow slides, then chromosomal and genetic data are usually received by doctors after the initial therapy. It is very important to understand the significance of all laboratory data and to integrate all results not only for diagnosis but also for management of patients with AML. PMID- 11064590 TI - [Total hematological analysis system as a screening test for hematological malignancy]. AB - Measurement of complete blood cell count and white blood cell differentiation is an essential laboratory test and the most important screening test for hematological malignancy. Recently, several automated blood cell analyzers have been developed to improve accuracy and precision. When flag messages generated in the presence of morphological abnormalities of the samples are displayed, manual revision is necessary. In our laboratory, the manual revision rate has been 35 40%. Therefore blood cell analyzers are useful in screening for abnormalities as well as greatly reducing expensive and time-consuming manual differential procedures. In addition, automated blood cell analyzers can provide several types of useful information including the leukocyte distribution scattergram. However, most such information is not utilized in the clinical field. In the future, a total hematological analysis system will be constructed so that all information provided by automated blood cell analyzer and by manual methods are available. PMID- 11064591 TI - [Morphology and immunophenotyping in hematological malignancies]. AB - We evaluated 443 outpatients and inpatients in Keio University Hospital between 1994 and 1999. Morphologic features from peripheral blood and bone marrow aspiration were evaluated in our hematology laboratory, using Wright-Giemsa, peroxidase staining films and other cytochemistry. Immunophenotype was determined by cell surface antigen analysis by laser flow cytometry, FACscan, using various monoclonal antibodies. Information on cytogenetic and molecular genetic characteristics can be also integrated for diagnosis. One hundred fifty patients were diagnosed with acute leukemia, in which 59 cases were ALL and 91 cases were AML. Seventy-four cases were MDS, 76 cases were myeloproliferative disorders, 21 cases were CLL related disorders, 104 patients were malignant lymphoma, and 18 cases were multiple myeloma. The ratio of male to female was 1.7. The probability of diagnostic rate by Immunophenotyping was estimated by Discriminant analysis in 189 patients, using multivariate analysis of immunophenotype compared to morphology. The average probability by immunophenotypic analysis for diagnostic rate was 91.7%, in which the probability for NHL was very high of 97.1%. Thus, morphologic and immunophenotypic analysis is most essential and basic approach in laboratory hematology, from the perspective of rapid and precise diagnostic methods. Recent advance appreciates the rapid contribution for diagnosis by immunophenotypic analysis. Furthermore, Tele-hematology would contribute the standardization for morphologic method in the near future. PMID- 11064592 TI - [Southern blot hybridization analysis for lymphoid neoplasms]. AB - The recent remarkable progress of molecular biology gave us new tools for examination of the etiology and pathology of many diseases. Hence, the introduction of such techniques in our clinical laboratory area is becoming more important and indispensable year by year. In addition to morphological and phenotypical analysis, we are now establishing a molecular diagnosis system to support the clinical diagnosis of hematological malignancies. In the present study, we show our ongoing results focusing on adult T-cell leukemia(ATL). Southern blot analysis for detection of the monoclonal integration of HTLV-I proviral genome, a causative agent of ATL, was performed using digoxigenin labeled probe. The sensitivity was equivalent to RI-labeled probe and was enough for clinical use. We are also doing Southern blot analysis for immunoglobulin heavy chain gene and T-cell receptor beta chain gene as custom service for the diagnosis of B-lymphoproliferative disorders and non-ATL T-lymphoproliferative disorders. We are reporting such results to the clinical site with our interpretation. Since the band size of the HTLV-I provirus obtained by Southern blot differed in each case and was sometimes smaller than the size of the complete virus, we further analyzed the structure of HTLV-I using long and accurate PCR(LA-PCR) and sequence-target-site PCR(STS-PCR). We found that the integrated HTLV-I provirus is divided into two subtypes, complete virus and defective virus. The incidence of defective virus was high in acute-type ATL compared with that in chronic- or lymphoma-type ATL. Although there are several difficulties in introducing such molecular analyses into the clinical laboratory area, we think that the need is increasing together with the progression of gene therapy and evidence-based medicine. PMID- 11064593 TI - [The genetic diagnosis of hematopoietic malignancy by polymerase chain reaction method]. AB - Since 1994, chimeric gene test for major-BCR/ABL, minor-BCR/ABL, AML1/MTG8, PML/RARa, DEK/CAN, CBFb/MYH11, MLL/LTG9, E2A/PBX1, TEL/AML1, and MLL/LTG4 by reverse transcription-nested-polymerase chain reaction (RT-nested-PCR) have been routinely performed in our laboratory. The frequently requested tests were those for major-BCR/ABL, minor-BCR/ABL, AML1/MTG8, PML1/RARa, and TEL/AML1 chimeric genes, being the test for major-BCR/ABL the most frequent one, accounting for more than 50% of the total orders. By the high sensitivity for minimal residual disease(MRD) detection, these methods are extremely useful for the therapeutic control of chemotherapy and bone marrow transplantation, as well as for prediction of prognosis. However, in the most cases a well-controlled monitoring could not be obtained, due to the 6-months interval between the tests, the minimal interval allowed by the insurance. The following points should be carefully observed. (1) The setting site of the primer can affect the sensitivity and the specificity of the test; (2) For the detection of chimeric genes with multiple translocation breakpoints, a genetic DNA sequencing is necessary to confirm them; (3) In patients receiving chemotherapy or bone marrow transplantation, that show extremely low blood cell counts, false-negative tests dependent on the quality of the extracted RNA and the low volume of DNA, should be avoided by careful managements. For improvements of the MRD detection tests, the quantification of the expression levels of chimeric gene and WT1 mRNA is necessary. PMID- 11064594 TI - [Inflammatory markers, especially the mechanism of increased CRP]. AB - Acute phase proteins are synthesized mainly in the liver cells, induced by various inflammatory cytokines which are produced by activated macrophages/monocytes at the inflammatory sites. C-reactive protein is a principal acute phase protein, and increased most significantly upon various inflammations. False negative results may be recognized in the patients with viral infections, collagen diseases such as SLE, PSS, dermatomyositis, ulcerative colitis, Sjogren's syndrome, leukemia, cerebral infarction, etc. PMID- 11064595 TI - [Exploratory analysis of elevated C-reactive protein without leukocytosis from the clinical laboratory database]. AB - We studied the characteristics of admitted patients who showed discrepancy between C-reactive protein(CRP) and white blood cell count(WBC). We extracted those patients from our laboratory information system by two criteria: WBC is less than 9500/microliter and either(1) CRP is more than 5.0 mg/dl, or(2) the pair of CRP and WBC is out of 95% confidence ellipse. We found 346 and 90 cases by the two criteria, respectively. They consisted of a variety of diseases, prevalent were such as pneumonia, rheumatoid arthritis, malignant lymphoma, post operative state and so on by either criterion. There was predominance of elderly patients as a whole. The analysis of individual time courses revealed that WBC did not change in parallel with CRP in patients with rheumatoid arthritis and malignant lymphoma, while they paralleled in those with infectious diseases and post-operation states. The elevation of WBC in some patients might have been overlooked since WBC was not always to be ordered together with CRP. We need a prospective study to closely analyze serial relationship between CRP and WBC for factors leading to the discovery. PMID- 11064596 TI - [A case of hemophagocytosis syndrome with excessive Fe and ferritin concentration in serum and with a good prognosis]. AB - Investigation in the present case began with a high serum Fe concentration. The routine method for determining serum Fe concentration in our hospital is direct photoscopic assay. When we assayed serum Fe concentration by a deproteinization method, it showed a higher value. We also measured serum ferritin concentration, and suspected that high ferritin interfered with serum Fe concentration. We checked the dose-dependent effect of ferritin on serum Fe determination. The present case, a 64-year-old female, showed a reduction of Plt count, and an elevation of LDH, CRP and so on, meeting most of the criteria for hemophagocytosis syndrome. Bone marrow smears revealed phagocytosis of blood cells. The present case had been followed for rheumatoid arthritis and medicated with oral iron pills. In her liver and spleen, high amounts of Fe had accumulated, and excessive Fe was being released into the blood by any trigger, such as infection, stress, cytokine activation, and/or HPS. PMID- 11064597 TI - [Cases of hemophagocytic syndrome in Tenri Hospital]. AB - The author reviewed clinical and laboratory findings included white blood cells(WBC), C-reactive protein(CRP) in cases of hemophagocytic syndrome(HPS). MATERIALS: Eleven cases of HPS treated in Tenri hospital between 1985 and 1999 were studied. HPS was diagnosed by clinical and laboratory findings. There were 6 children(9 months-5 years-old, male 2: female 4) and 5 adults(31-74 years-old, all: female). RESULTS AND DISCUSSION: Etiology of HPS was viral infection in 8 cases and autoimmune diseases, drug allergy, malignant lymphoma in the other 3 cases. Clinical and laboratory findings that indicated poor prognosis were hepatosplenomegaly complicated with lymphadenopathy, disseminated intravascular coagulopathy, increase of alpha 1-globulin and hyperbilirubinemia. The number of WBC ranged between 500 and 4300/microliter. WBC was decreased in 9 cases and there were cases demonstrating relative lymphopenia or neutropenia. However, CRP increased in all cases (0.2-30.7 mg/dl), although in 8 cases, the CRP values were less than 4 mg/dl. There was no relation between the number of WBC and CRP values and neither of these items showed any relation to hepatosplenomegaly or values of lactate dehydrogenase and ferritin. We should prepare histological sections along with smears from bone marrow examination to increase the sensitivity of diagnosing HPS. We should also note that even if there is no increase in hemophagocytes, we can not deny HPS. CONCLUSION: Cytopenia and increases of CRP, alpha 1-globulin, lactate dehydrogenase and ferritin were common findings in HPS, but there was no relation among these findings and there were large differences in these findings in each case. PMID- 11064598 TI - [Hemophagocytic syndrome: problems in diagnosis]. AB - Hemophagocytic syndrome(HPS) is characterized by high-grade fever, pancytopenia, liver dysfunction and proliferation of benign histiocytes with hemophagocytosis. These clinical manifestations result from the overproduction of cytokines including interferon-gamma, interleukin-2, tumor necrosis factor-alpha by activated T cells(Th1 cells) and macrophages. HPS is divided into a hereditary/primary form and a secondary form. The latter is further classified into various entities by triggering factors and underlying diseases. The multiplicity of this syndrome makes it difficult to diagnose by a simple criteria. This paper gives a overview of HPS and describes the problems of diagnosing each sub-entity of HPS. PMID- 11064599 TI - [A consideration on discrepancy of results obtained between commercial CRP measurement kits]. AB - We investigated the frequency of the occurrence of discrepancies using various patients' sera between several commercial C reactive protein(CRP) kits with turbidimetric immunoassay, and the elucidation of the mechanism of the non specific reaction, also investigated methods of minimizing false results in some procedures. CRP was measured in 79 patients' sera containing monoclonal immunoglobulins [Multiple Myeloma: 45 patients' sera(IgG type, 37; IgA type, 7; BJP type, 1); Waldenstrom's disease: 8; Benign M proteinemia: 26], 70 patients' sera with polyclonal high gamma-globulinemia (> 2.0 g/dl) and 91 patients' sera with positive rheumatoid factor. Two different patients' sera, one(chronic hepatitis C) with Waldenstrom's disease and the other(purpura) with polyclonal high gamma-globulinemia, showed marked discrepancies. We found these discrepancies were induced by milky turbidity produced by non-specific reaction between high molecular weight components(cryoglobulin composed from IgM-IgG in the Waldenstrom case and immune complex in the polyclonal high gamma-globulinemia case), and the buffer solution contained polyethylene glycol(PEG) as a catalyst. The discrepancy was minimized by the following procedures: (a) decreasing the PEG concentration, (b) increasing pH of the buffer solution, (c) using a small volume of the secondary reagent(R2), and (d) using rabbit anti-human CRP serum instead of goat serum. PMID- 11064600 TI - [Interference in turbidimetric immunoassay for serum C-reactive protein due to serum protein abnormalities an immune complex and rheumatoid factor]. AB - Interference in immunoassays for CRP caused by serum protein abnormalities was studied with special reference to turbidimetric immunoassay(TIA) and latex photometric immunoassay(LPIA). One of the interfering factors in TIA is immune complex or agglutinating immunoglobulin which reacts with a chemical component like polyethylene glycol in reagent of the first reaction and causes remarkable turbidity in the initial phase. Since the turbidity decreases gradually and can not be eliminated within the first reaction, the second reaction is affected by the continuing reduction in absorbance, resulting in falsely low CRP values. As examples of this kind of interference, two cases, case 1 and 2 were presented. Case 1 involved malignant lymphoma with paraproteinemia of monoclonal IgA(kappa) and Case 2 was chronic viral hepatitis type C with type II cryoglobulinemia composed of monoclonal IgM(kappa) and polyclonal IgG. Rheumatoid factor(RF) is another factor that interferes in TIA which reacts with antibody in the reagent of the second reaction and causes falsely high CRP values. The effect of RF is especially remarkable in LPIA. As an example, a case of chronic rheumatoid arthritis(Case 3) with polyclonal IgM is presented. Predilution of serum(5-fold dilution with physiological saline solution) was proved to be effective in reducing the intensity of interference in TIA caused by immune complex or agglutinating immunoglobulin in Cases 1 and 2. Covalent binding of IgG(Fab')2 to latex particle instead of physical adsorption of IgG was shown to be efficient in eliminating interference caused by RF in LPIA in Case 3. PMID- 11064601 TI - [Factors interfering with CRP assay (from the manufacturer's perspective)]. AB - Recently, there have been many highly sensitive methods for CRP assay. However, there are also many factors that interfere with CRP assay. We are taking measures to avoid interference due to factors such as, myeloma protein, rheumatoid factor, lipaemia(a substance similar to detergent in the capillary tube, etc). Currently CRP assay methods which are not influenced by these factors are available. However, other unknown factors that interfere with CRP assay have become apparent. We think that these are peculiar to the immunoassay. We are taking measures to identify these interference factors and resolve the problem. In the following, we will explain some experiments that will protect our re-agent from these interference factors. PMID- 11064602 TI - West meets east-looking for the interphase of Western medicine and traditional oriental medicine in future. PMID- 11064603 TI - [Comparison between leucyl aminopeptidase and pseudo leucine aminopeptidase activities in sera]. AB - The highest activities of leucyl aminopeptidase(LAP, cytosol aminopeptidase, EC 3.4.11.1) in sera have been found in patients with acute hepatitis(Kanno et al., Am J Clin Path, 82: 700-705, 1984). I observed inpatients with very high activities of LAP and alcohol dehydrogenase(AD) in sera. However, only slight elevations of serum pseudo leucine aminopeptidase(PLA), that is, membrane alanyl aminopeptidase(MAA, microsomal aminopeptidase, EC 3.4.11.2) activities for hydrolysis of leucyl-4-nitroanilide were observed in these patients. They were patients in critical care unit with ischemia caused by a cardiopulmonary arrest, multiple trauma, acute myocardial infarction or operation. Therefore, we should measure LAP activities in sera rather than PLA(MAA) activities in these patients. PMID- 11064604 TI - The utility of pulmonary artery catheterization. PMID- 11064605 TI - Airway devices: where now and where to? PMID- 11064606 TI - Ropivacaine pharmacokinetics after caudal block in 1-8 year old children. AB - We studied the pharmacokinetics after caudal block of ropivacaine (2 mg ml-1, 1 ml kg-1) performed in 20 children aged 1-8 yr undergoing subumbilical surgery, in this open, non-comparative, multicentre study. Venous blood samples were collected up to 12-36 h. The mean (SD) peak plasma concentration, 0.47 (0.16) mg litre-1, was achieved after 12-249 min. The free fraction was 5% and the highest individual peak plasma concentration of free ropivacaine was 0.04 mg litre-1. Clearance was 7.4 (1.9) ml min-1 kg-1 and the terminal half-life 3.2 (0.8) h. Thus, the free plasma concentrations of ropivacaine were well below those associated with toxic symptoms in adults and the capacity to eliminate ropivacaine seems to be well developed in this age group. In this open study of 20 patients, ropivacaine was well tolerated and provided satisfactory postoperative pain relief without observable motor block. PMID- 11064607 TI - Pharmacokinetics of rectal paracetamol after repeated dosing in children. AB - Twenty-three children (aged between 9 weeks and 11 yr) were given paracetamol suppositories 25 mg kg-1 every 6 h (maximum 5 days) after major surgery and serum and saliva concentrations were measured. There was a good correlation (r = 0.91, P < 0.05) between saliva and serum concentrations. A one-compartment linear model with first-order elimination and absorption and lag-time was fitted to the data (ADAPT II). At steady state, the mean (SD) concentration was 15.2 (6.8) mg litre 1. Mean (SD) time to reach 90% of the steady state concentration was 11.4 (8.6) h. Body weight, age and body surface area were well correlated (P < 0.05) with clearance and apparent volume of distribution. There was no evidence of accumulation leading to supratherapeutic concentrations during this dosing schedule for a mean of approximately 2-3 days. PMID- 11064608 TI - Peripheral lidocaine but not ketamine inhibits capsaicin-induced hyperalgesia in humans. AB - We examined the effect of the subcutaneous infiltration of ketamine, lidocaine and saline before injury on capsaicin-induced pain and hyperalgesia. Twelve healthy volunteers participated in two separate, randomized, double-blind, placebo-controlled crossover experiments. In experiment 1, 100 micrograms capsaicin was injected intradermally in one volar forearm 10 min after the skin had been pretreated with lidocaine 20.0 mg in 2.0 ml or 0.9% saline 2.0 ml at the capsaicin injection site. In experiment 2, a similar capsaicin test was given 10 min after the skin had been pretreated with ketamine 5 mg in 2.0 ml or 0.9% saline 2.0 ml. To control for possible systemic effects, the capsaicin injection site was pretreated by injection of saline into the skin and the contralateral arm was treated with active drug, and vice versa. Outcome measures were spontaneous pain, pain evoked by punctate and brush stimuli, and areas of brush evoked and punctate-evoked hyperalgesia. Lidocaine reduced all measures compared with placebo (P < 0.001), whereas ketamine failed to change any measures. Pain scores and areas of hyperalgesia were not affected when the contralateral site was infiltrated with ketamine or lidocaine. Lidocaine produced no side-effects, whereas ketamine produced paraesthesia, dizziness and sleepiness in six out of 24 (25%) cases. Blocking peripheral sodium channels with locally administered lidocaine reduces spontaneous pain and capsaicin-induced hyperalgesia but local block with the NMDA-type glutamate receptor antagonist ketamine has no effect on capsaicin-induced pain and hyperalgesia. PMID- 11064609 TI - Remifentanil inhibits muscular more than cutaneous pain in humans. AB - In experimental studies, drug-induced analgesia is usually assessed by cutaneous stimulation. If analgesics act differently on cutaneous and deep nociception, the results of these studies may not be entirely applicable to clinical pain involving deep structures. We tested the hypothesis that opioids have different abilities to inhibit cutaneous and muscular pain. Either the opioid remifentanil or placebo was infused in 12 healthy volunteers in a cross-over fashion. Repeated electrical stimulation (five impulses at 2 Hz) was applied to both skin and muscle. Pain thresholds were recorded. Remifentanil caused a higher increase in the muscular pain thresholds than in the cutaneous pain thresholds (P = 0.035). We conclude that opioids inhibit muscular pain more strongly than cutaneous pain in humans. PMID- 11064610 TI - Comparison of alfentanil, fentanyl and sufentanil for total intravenous anaesthesia with propofol in patients undergoing coronary artery bypass surgery. AB - We have studied the pharmacokinetics and pharmacodynamics of alfentanil, fentanyl and sufentanil together with propofol in patients undergoing coronary artery bypass graft surgery (CABG). Sixty patients (age 40-73 yr, 56 male) were assigned randomly to receive alfentanil, fentanyl or sufentanil and propofol. Plasma concentrations of these drugs and times for the plasma concentration to decrease by 50% (t50) and 80% (t80) after cessation of the infusion were determined. Times were recorded to awakening and tracheal extubation. Total dose and plasma concentrations of propofol were similar in all groups. Mean total doses of alfentanil, fentanyl and sufentanil were 443, 45 and 4.4 micrograms kg-1, respectively. Time to awakening did not differ significantly. In patients receiving fentanyl, the trachea was extubated on average 2 h later than in those receiving sufentanil and 3 h later than in those receiving alfentanil (P < 0.05). The t80 of fentanyl was longer (P < 0.05) than that of alfentanil or sufentanil, and there was a linear correlation between the t80 of the opioid and the time to tracheal extubation (r = 0.51; P < 0.01). However, the t50 values for these opioids were similar and did not correlate with recovery time. In conclusion, patients undergoing CABG and who were anaesthetized with fentanyl and propofol needed mechanical ventilatory support for a significantly longer time than those receiving alfentanil or sufentanil and propofol. On the basis of the interindividual variation observed, the time to tracheal extubation was most predictable in patients receiving alfentanil and most variable in patients receiving fentanyl, a finding which may be important if the patients are transferred to a step-down unit on the evening of the operation. PMID- 11064611 TI - Clinical comparison of 'single agent' anaesthesia with sevoflurane versus target controlled infusion of propofol. AB - The introduction of total intravenous anaesthesia (TIVA) and the use of volatile induction/maintenance anaesthesia (VIMA) has led to the rediscovery of 'single agent' anaesthesia, eliminating the transition phase from induction to maintenance. We compared quality, patient acceptability and cost of TIVA using target control infusion (TCI) with propofol and VIMA with sevoflurane. Forty patients undergoing spinal surgery of 1-3 h were assigned to one of two groups. Group I received propofol-air-oxygen for induction followed by propofol-air oxygen for maintenance. Group II received 8% sevoflurane-oxygen for induction and sevoflurane-oxygen-nitrous oxide for maintenance. Propofol had a significantly faster mean (SD) induction time (67 (20) s) than sevoflurane (97 (38) s) but was associated with double the incidence of involuntary movements. Although not significant, twice the number of interventions by the anaesthetist were required to maintain an adequate level of anaesthesia in the sevoflurane group. Emergence times, characteristics, postoperative nausea, vomiting and pain were unaffected by the anaesthetic technique. However, a more predictable emergence time was found following sevoflurane. Cardiovascular stability was good and comparable in both groups. The majority of patients found either technique acceptable and would choose the same anaesthetic again. Induction and maintenance was substantially cheaper with sevoflurane (28.06 Pounds) compared with propofol (41.43 Pounds). PMID- 11064612 TI - A series of thyroplasty cases under general anaesthesia. AB - Thyroplasty is an operation on the upper airway to improve voice quality in patients with unilateral vocal cord paralysis. It requires access to an uninstrumented larynx and a functional assessment of vocal cord medialization. It is a difficult anaesthetic procedure that requires sharing the airway with the surgeon. We describe an anaesthetic technique to give good operating conditions and a safe airway, using total intravenous anaesthesia, a laryngeal mask airway and intraoperative fibreoptic endoscopic assessment of the larynx, and present a series of 13 patients. Other anaesthetic techniques for thyroplasty are described and discussed. PMID- 11064613 TI - Perioperative blood salvage during surgical correction of craniosynostosis in infants. AB - Surgical correction of craniosynostosis in infants is a very haemorrhagic procedure. The aim of this study was to determine whether the perioperative use of the continuous autotransfusion system (CATS) would reduce homologous transfusion during repair of craniosynostosis. Two groups of patients were studied according to the availability of the CATS in our hospital. The control group had surgery before the system was introduced and the study group had operations subsequently. Use of CATS was associated with a significant decrease in the median (95% confidence interval) volume of homologous blood transfused [413 (250-540) ml in the control group versus 317 (150-410) ml in the CATS group, P = 0.02] and in the median (95% confidence interval) number of packed red cell units transfused [2 (1-2) in the control group versus 1 (1-2) in the CATS group, P = 0.04] in the perioperative period. Use of CATS is associated with a reduction in homologous transfusion during the surgical correction of craniosynostosis in infants. PMID- 11064614 TI - Publications on paediatric anaesthesia: a quantitative analysis of publication activity and international recognition. AB - A comprehensive compilation of the current international literature on paediatric anaesthesia is lacking. The aim of this study was to identify all articles on clinical practice in paediatric anaesthesia, to name the respective journals, and to assess the publication activity and international recognition of selected countries for a 6-yr period (1993-1998). The search comprised an article-to article evaluation ('hand search') of 12 peer-reviewed anaesthesia journals, as well as an Internet-based ('SilverPlatter') Medline-search (3,900 medical journals, US National Library of Medicine), both limited to original articles, case reports, reviews and editorials. Selected physical characteristics, for example the number of infants and children aged 0-14 yr old, the number of anaesthetists (specialists) and current impact factors (Science Citation Index) served to assess publication activity and international recognition. During the time period studied, 2259 articles (377/yr) were published on paediatric anaesthesia in 295 medical journals. The articles were primarily written in English (85.1%) and the majority originated from the USA (35.4%) and the UK (12.6%). The largest number of publications (77.7%) appeared in 29 anaesthesia journals, all referenced in Medline, with 46% being published by only five journals. Most authors published in journals of their home country/region. Authors from the UK ranked highest in publication activity, followed by those from Canada, Switzerland, Sweden and Denmark. The highest impact factor was achieved by US and UK authors. We conclude that publications on paediatric anaesthesia are clustered in a small number of journals and are written predominantly by authors from English-speaking countries, who achieved the highest international recognition. PMID- 11064615 TI - Gastric mucosal end-tidal PCO2 difference as a continuous indicator of splanchnic perfusion. AB - Gastric mucosal and arterial blood PCO2 must be known to assess mucosal perfusion by means of gastric tonometry. As end-tidal PCO2 (PE'CO2) is a function of arterial PCO2, the gradient between PE'CO2 and gastric mucosal PCO2 may reflect mucosal perfusion. We studied the agreement between two methods to monitor gut perfusion. We measured the difference between gastric mucosal PCO2 (air tonometry) and PE'CO2 (= DPCO2gas) and the difference between gastric mucosal PCO2 (saline tonometry) and arterial blood PCO2 (= DPCO2sal) in 20 patients with or without lung injury. DPCO2gas was greater than DPCO2sal but changes in DPCO2gas reflected changes in DPCO2sal. The bias between DPCO2gas and DPCO2sal was 0.85 kPa and precision 1.25 kPa. The disagreement between DPCO2gas and DPCO2sal increased with increasing dead space. We propose that the disagreement between the two methods studied may not be clinically important and that DPCO2gas may be a method for continuous estimation of splanchnic perfusion. PMID- 11064616 TI - Inhibition of nitric oxide synthesis augments pulmonary oedema in isolated perfused rabbit lung. AB - The role of nitric oxide (NO) in precipitating pulmonary oedema in acute lung injury remains unclear. We have investigated the mechanism of involvement of NO in the maintenance of liquid balance in the isolated rabbit lung. Thirty pairs of lungs were perfused with colloid for up to 6 h, during which pulmonary vascular resistance (PVR) and capillary pressure (PCP) were measured frequently, and time to gain 5 g in weight (t5) was recorded. Four protocols with different perfusate additives were studied: (i) none (control, n = 11); (ii) 10 mmol NG-nitro-L arginine methyl ester (L-NAME) (n = 6); (iii) 10 mmol L-NAME with 100 mumol lodoxamide, an inhibitor of mast cell degranulation (n = 7); (iv) 10 mmol L-NAME with 10 mumol 8-bromo-3',5'-cyclic guanosine monophosphate (8Br-cGMP), an analogue of cGMP that may reduce vascular permeability by relaxing contractile elements in endothelial cells (n = 6). Neither PVR nor PCP differed between protocols. L-NAME markedly reduced t5 from 248 (27) min (mean (SEM)) in protocol (i) to 144 (5) min in protocol (ii) (P < 0.05). Both lodoxamide (t5 = 178 (7) min) and 8Br-cGMP (t5 = 204 (10) min) substantially corrected the effect of L NAME (P < 0.005). Results suggest that maintenance of a low permeability by NO may involve mast cell stabilization and endothelial cell relaxation. PMID- 11064617 TI - Static versus dynamic respiratory mechanics for setting the ventilator. AB - The lower inflection point (LIP) of the inspiratory limb of a static pressure volume (PV) loop is assumed to indicate the pressure at which most lung units are recruited. The LIP is determined by a static manoeuvre with a PV-history that is different from the PV-history of the actual ventilation. In nine surfactant deficient piglets, information to allow setting PEEP and VT was obtained, both from the PV-curve and also during ongoing ventilation from the dynamic compliance relationship. According to LIP, PEEP was set at 20 (95% confidence interval 17 22) cm H2O. Volume-dependent dynamic compliance suggested a PEEP reduction (to 15 (13-18) cm H2O). Pulmonary gas exchange remained satisfactory and this change resulted in reduced mechanical stress on the respiratory system, indirectly indicated by volume-dependent compliance being consistently great during the entire inspiration. PMID- 11064618 TI - Morphine tolerance increases [3H]MK-801 binding affinity and constitutive neuronal nitric oxide synthase expression in rat spinal cord. AB - N-Methyl-D-aspartate (NMDA) receptor antagonists and nitric oxide synthase (NOS) inhibitors inhibit morphine tolerance. In the present study, a lumbar subarachnoid polyethylene (PE10) catheter was implanted for drug administration to study alterations in NMDA receptor activity and NOS protein expression in a morphine-tolerant rat spinal model. Antinociceptive tolerance was induced by intrathecal (i.t.) morphine infusion (10 micrograms h-1) for 5 days. Co administered (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate (MK-801) (10 micrograms h-1 i.t.) with morphine was used to inhibit the development of morphine tolerance. Lumbar spinal cord segments were removed and prepared for [3H]MK-801 binding assays and NOS western blotting. The binding affinity of [3H]MK-801 was higher in spinal cords of morphine-tolerant rats (mean (SEM) KD = 0.41 (0.09) nM) than in control rats (1.50 (0.13) nM). There was no difference in Bmax. Western blot analysis showed that constitutive expression of neuronal NOS (nNOS) protein in the morphine-tolerant group was twice that in the control group. This up-regulation was partially prevented by MK-801. The results suggest that morphine tolerance affects NMDA receptor binding activity and increases nNOS expression in the rat spinal cord. PMID- 11064619 TI - Determination of succinylcholine in plasma by high-pressure liquid chromatography with electrochemical detection. AB - The plasma concentration of the neuromuscular blocking drug, succinylcholine, is difficult to measure. We have measured concentrations of the breakdown product of succinylcholine, choline, to assess whether choline concentration gives an accurate measure of succinylcholine concentration. Choline concentration was measured by HPLC and electrochemical detection in two blood or plasma samples, one in which succinylcholine hydrolysis was inhibited by 10(-5) M physostigmine and another in which succinylcholine was completely hydrolysed in 20 min by 200 mU butyrylcholinesterase at 37 degrees C. The difference in choline content between the two samples gives the succinylcholine concentration. Ninety-five per cent recovery of choline was achieved. Choline standard curves were linear from 156 pmol ml-1 to 200 nmol ml-1. Within-day and between-day mean coefficients of variation for succinylcholine hydrolysis were small (mean (SD) 3.7% (1.2%) and 3.8% (1.6%), respectively). We conclude that this method of measuring succinylcholine concentration in blood is accurate, repeatable and relatively easy. PMID- 11064620 TI - The role of albumin in critical illness. PMID- 11064621 TI - Pulmonary artery catheterization and mortality in critically ill patients. AB - Pulmonary artery catheters are widely used in intensive care, but evidence to support their widespread use in sparse. Some published data suggest that greater mortality is associated with use of these catheters. The largest study to date looked at > 5500 patients in several centres in America and found a greater 30 day mortality in those patients receiving a pulmonary artery catheter. We tested the hypothesis that, on our intensive care unit, mortality was greater for those patients receiving a pulmonary artery catheter. Using a propensity score to account for severity of illness, the odds ratio for mortality in those patients receiving a pulmonary artery catheter was 1.08 (95% confidence interval 0.87 1.33). We believe that continued use of the pulmonary artery catheter is safe; a large randomized controlled trial examining outcome is unlikely to provide an adequate answer. PMID- 11064622 TI - Fentanyl versus sufentanil: plasma concentrations during continuous epidural postoperative infusion in children. AB - No pharmacokinetic data are available with respect to the plasma concentrations and fentanyl or sufentanil during epidural administration in children. This double-blind randomized study included 12 children (5-12 yr). Patients in group F were given an epidural loading dose of fentanyl 1.5 micrograms kg-1 and in group S sufentanil 0.6 microgram kg-1. Both groups then received a continuous epidural infusion of bupivacaine 5 mg kg-1 day-1 with either fentanyl 5 micrograms kg-1 day-1 or sufentanil 2 micrograms kg-1 day-1. An epidural PCA system was also given to the children (bolus: bupivacaine 0.2 mg kg-1 and fentanyl 0.2 microgram kg-1 or sufentanil 0.08 microgram kg-1). Maximal median concentrations of plasma (0.117-0.247 ng ml-1 for fentanyl and 0.027-0.074 ng ml-1 for sufentanil) were reached approximately 30 and 20 min respectively after the loading doses. These values were similar to those measured after 48 h. PMID- 11064623 TI - Peribulbar anaesthesia with 1% ropivacaine and hyaluronidase 300 IU ml-1: comparison with 0.5% bupivacaine/2% lidocaine and hyaluronidase 50 IU ml-1. AB - The low toxicity of ropivacaine makes it attractive for peribulbar anaesthesia. However, its motor-sparing properties are undesirable when akinesia is important. Hyaluronidase (300 IU ml-1) promotes the onset and quality of peribulbar blockade when used with other agents. We investigated the onset and quality of ocular akinesia in 80 patients randomized to receive 1% ropivacaine plus hyaluronidase 300 IU ml-1 (group 1), or bupivacaine 0.5%/Lidocaine 2% plus 50 IU ml-1 hyaluronidase (group 2). Ocular akinesia was scored from 0 (no movement) to 8 (full movement) every 2 min for 20 min. The groups showed no difference in the rate of onset or degree of akinesia achieved (analysis of variance with repeated measures; P = 0.34). Sixty per cent of patients in group 1 and 55% in group 2 achieved akinesia scores of < or = 4 by 6 min (chi 2 test; P = 0.5). We conclude that both peribulbar solutions produce equivalent onset and quality of ocular akinesia. PMID- 11064624 TI - Lack of pre-emptive analgesic effects of local anaesthetics on neuropathic pain. AB - We investigated the significance of pre-emptive analgesia using a well-known model of neuropathic pain in rats. Lignocaine, bupivacaine or saline was applied locally to the left L5-L6 spinal nerve before or 4 days after nerve injury. Mechanical allodynia was then evaluated before and after injury. Pre- and post injury treatment with local anaesthetics both resulted in a two- to threefold increase in the pain threshold, as manifested by a significant increase in von Frey measurements. However, this effect lasted only 24 h. Our study in rats questions the beneficial effect of a single dose of local anaesthetic as pre emptive analgesia. PMID- 11064625 TI - Comparison of intubating conditions following propofol and succinylcholine with propofol and remifentanil 2 micrograms kg-1 or 4 micrograms kg-1. AB - We evaluated the intubating conditions, haemodynamic responses and duration of apnoea in 60 healthy adult patients after propofol 2 mg kg-1 combined with either a bolus of remifentanil 2 micrograms kg-1 or 4 micrograms kg-1, or succinylcholine 1 mg kg-1. Patients intubated following remifentanil showed dose dependent intubating conditions, similar at 4 micrograms kg-1 to the conditions produced with succinylcholine. Post-induction mean arterial pressure decreased from baseline values by 21% (P < 0.0001), 28% (P < 0.0001) and 8% (P > 0.05) in the remifentanil 2 micrograms kg-1, remifentanil 4 micrograms kg-1 and succinylcholine 1 mg kg-1 groups, respectively. The mean (SD) duration of apnoea following induction was 9.3 (2.6) min and 12.8 (2.9) min in the remifentanil 2 micrograms kg-1 and 4 micrograms kg-1 groups, and 6.0 (0.9) min in the succinylcholine group (P < 0.001 between groups). PMID- 11064626 TI - Two cases of difficult intubation managed by a handmade device. AB - The cuffed pharyngeal tube (CPT) is a hand-made device designed to maintain upper airway patency during anaesthesia. Two cases of difficult intubation managed successfully using the new device are described. Insertion was easily achieved at the first attempt in each case. The incisor-tip distance of the CPT is 14 cm and the cuff is inflated with 60 ml of air in an average adult. PMID- 11064627 TI - Use of nitric oxide for decompensated right ventricular failure and circulatory shock after cardiac arrest. AB - We describe a case of peri-operative cardiac arrest, severe right ventricular failure and pulmonary hypertension in a 60-yr-old woman with interstitial pulmonary fibrosis. Inhaled nitric oxide therapy rapidly improved arterial oxygenation and haemodynamic variables, allowing recovery and weaning from mechanical ventilation. Subsequently, the patient was discharged from the cardiac intensive care unit. PMID- 11064628 TI - Complete recovery of consciousness in a patient with decorticate rigidity following cardiac arrest after thoracic epidural injection. AB - A 46-yr-old man with dysaesthesia (burning sensation) following herpes zoster in the left upper chest region was treated with a single thoracic (T2/T3) epidural injection (1.0% lidocaine 3 ml + 0.125% bupivacaine 3 ml) as an outpatient. Twenty minutes after the injection, a nurse noticed the patient to be unconscious with dilated pupils, apnoea and cardiac arrest. Following immediate cardiopulmonary resuscitation, the patient was treated with an i.v. infusion of thiamylal sodium 2-4 mg kg-1 h-1 and his lungs were mechanically ventilated. When the patient developed a characteristic decorticate posture, mild hypothermia (oesophageal temperature, 33-34 degrees C) was induced. On the 17th day of this treatment, after rewarming (35.5 degrees C) and discontinuation of the barbiturate, the patient responded to command. Weaning from the ventilator was successful on the 18th day. About 4 months after the incident, the patient was discharged with no apparent mental or motor disturbances. We suggest that mild hypothermia with barbiturate therapy may have contributed to the successful outcome in this case. PMID- 11064629 TI - Phaeochromocytoma diagnosed during labour. AB - The diagnosis of phaeochromocytoma during pregnancy is rare. We present the management of vaginal delivery in a woman diagnosed with the condition during labour. A Medline search and follow-up of references failed to find any similar report in the last 30 years. PMID- 11064630 TI - Hyperbaric oxygen therapy. PMID- 11064631 TI - Hyperbaric oxygen therapy. PMID- 11064632 TI - Rectal paracetamol has a significant morphine-sparing effect after hysterectomy. PMID- 11064633 TI - Sedation of children by non-anaesthetists. PMID- 11064634 TI - Sedation of children by non-anaesthetists. PMID- 11064635 TI - The interscalene approach to the cervical plexus. PMID- 11064636 TI - The interscalene approach to the cervical plexus. PMID- 11064637 TI - Nitric oxide concentration in the presence of potent inhalation anaesthetics. PMID- 11064638 TI - Anaesthesia, jugular venous oxyhaemoglobin saturation and coronary artery bypass surgery. PMID- 11064639 TI - Spinal cord injury and direct laryngoscopy--the legend lives on. PMID- 11064640 TI - Randomized controlled comparison of epidural bupivacaine versus pethidine for analgesia in labour. PMID- 11064641 TI - Propofol with and without nitrous oxide. PMID- 11064642 TI - Inhalation sedation with sevoflurane for upper gastrointestinal endoscopy. PMID- 11064643 TI - Radiation dosimetry using polymer gels: methods and applications. AB - New, complex radiotherapy delivery techniques require dosimeters that are able to measure complex three-dimensional dose distributions accurately and with good spatial resolution. Polymer gel is an emerging new dosimeter being applied to these challenges. The aim of this review is to present a practical overview of polymer gel dosimetry, including gel manufacture, imaging, calibration and application to radiotherapy verification. The dosimeters consist of a gel matrix within which is suspended a solution of acrylic molecules. These molecules polymerize upon exposure to radiation, with the degree of polymerization being proportional to absorbed dose. The polymer distribution can be measured in two or three dimensions using MRI or optical tomography and, after calibration, the images can be converted into radiation dose distributions. Manufacture of the gel is reported to be reproducible, and measured dose in the range 0-10 Gy is accurate to within 3-5%. In-plane image resolution of 1 mm x 1 mm, with image slice thicknesses of between 2-5 mm, is typically achievable using clinical 1.5 T MR scanners and standard T2 weighted imaging sequences. The gels have been used to verify a number of conventional and novel radiotherapy modalities, including brachytherapy, intensity modulated radiotherapy and stereotactic radiosurgery. All the studies have confirmed the value and versatility of the dosimetry technique. PMID- 11064644 TI - Rapidly growing small peripheral lung cancers detected by screening CT: correlation between radiological appearance and pathological features. AB - 12 peripheral small lung cancers (< 20 mm) of rapid growth (volume doubling time < 150 days), detected by repeated low dose CT screening, were evaluated to examine their CT features and to correlate such features with histopathological findings. Each patient's CT images, including follow-up and thin section CT images, were studied retrospectively to determine tumour growth rate and CT morphological features. Nine of the tumours exhibited a solid tumour growth pattern: seven of these showed a well defined, homogeneous, soft tissue density with spicular or lobulated margin. These seven tumours included small cell lung cancer (n = 3), moderately differentiated adenocarcinoma (n = 2), poorly differentiated adenocarcinoma (n = 1) and squamous cell carcinoma (n = 1). The other two tumours, a moderately differentiated adenocarcinoma and a well differentiated adenocarcinoma, appeared as irregular, soft tissue density nodules with poorly defined margins. The latter exhibited an air bronchogram pattern and a small cavity. The remaining three tumours exhibited a lepidic tumour growth pattern. They showed ground glass opacity or ground glass opacity with a higher density central zone on CT images and were well differentiated adenocarcinomas. In conclusion, most peripheral small lung cancers of rapid growth were adenocarcinomas. They also included small cell lung cancer and squamous cell carcinoma. The majority showed solid tumour growth pattern and lacked an air bronchogram and/or small air spaces in the nodule. Some well differentiated adenocarcinomas with lepidic tumour growth pattern also showed rapid growth. PMID- 11064645 TI - Is the appearance of microcalcifications on mammography useful in predicting histological grade of malignancy in ductal cancer in situ? AB - It has been proposed that the grade of malignancy of ductal carcinoma in situ (DCIS) of the breast can be estimated by the morphology of microcalcifications found on mammography. We correlated microcalcifications and histopathology in a retrospective blinded review. We reviewed all patients who underwent excisional breast biopsy over a 5 1/2-year period. Mammograms and pathology slides of all patients (n = 49) with DCIS of the breast were included in a blinded retrospective analysis. Mammographic microcalcifications were divided into four categories, "linear branching", "coarse granular", "fine granular" or "no microcalcification". Independently, pathology specimens were assigned to poorly, intermediately and well differentiated categories according to the consensus classification of DCIS introduced by the European Organisation for the Research and Treatment of Cancer. Two patients had no microcalcifications. 25 (53%) of the remaining 47 patients had "linear branching" microcalcifications, 10 (21%) had "coarse granular" and 12 (25.5%) had "fine granular" microcalcifications. 19 patients (40%) had poorly differentiated, 23 (49%) intermediately differentiated and 5 (11%) well differentiated subtypes of DCIS. 14 (56%) of the 25 patients with "linear branching" microcalcifications had poorly differentiated DCIS, 10 (40%) had intermediately differentiated and 1 (4%) had well differentiated DCIS. 3 (30%) of 10 patients with "coarse granular" microcalcifications had poorly differentiated DCIS, 5 (50%) had intermediately differentiated and 2 (20%) had well differentiated DCIS. 2 (17%) of 12 patients with "fine granular" microcalcifications had poorly differentiated DCIS, 8 (67%) had intermediately differentiated and 2 (17%) had well differentiated DCIS. These findings were not statistically significant. In conclusion, "linear branching" microcalcifications tended to be associated with higher pathological grading. However, correlation was poor and there was considerable overlap between categories. Histological type of DCIS cannot be predicted prospectively on mammographic appearances. PMID- 11064646 TI - The value of plain radiographs in the prediction of outcome in pelvic fractures treated with embolisation therapy. AB - 40 haemodynamically unstable patients with pelvic injuries were studied to assess the ability of plain radiographs to detect haemorrhagic sites. Pelvic radiographs and bilateral angiograms were reviewed separately for detection of haemorrhagic sites in both anterior and posterior segments. Sensitivity and specificity of pelvic radiographs for the detection of haemorrhagic sites were obtained by analysis of angiographic findings. Angiography demonstrated arterial injury in 106 divisions: 26 right anterior, 22 right posterior, 33 left anterior and 25 left posterior. Sensitivities of the radiographs for predicting haemorrhagic sites were higher in the anterior segment (right, 96%; left, 100%) than in the posterior segment (right, 73%; left, 83%). However, specificities were lower in the anterior segment (right, 79%; left, 78%) than in the posterior segment (right, 100%; left, 100%). In 15 (58%) of 26 patients with in-dwelling Foley catheters, including those with catheter deviation, the severity of anterior injury indicated on plain radiographs was correlated with angiographic findings. Plain radiographs of the pelvis proved useful for predicting haemorrhagic sites in haemodynamically unstable patients with pelvic fractures, especially in those with anterior fractures. PMID- 11064647 TI - Adynamic ileus after caesarean section mimicking intestinal obstruction: findings on abdominal radiographs. AB - The purpose of this study was to determine the spectrum of findings and the frequency of apparent distal colonic obstruction on abdominal radiographs in women with obstructive symptoms following Caesarean section. A search of radiology files yielded 21 patients who had abdominal radiographs because of obstructive symptoms during the early post-operative period. The radiographs were reviewed retrospectively to characterize the bowel gas patterns in these patients. Medical records were also reviewed to determine the treatment and patient course. Abdominal radiographs showed findings suggestive of distal colonic obstruction in 15 patients (71%), small bowel obstruction in 2 (10%), adynamic ileus in 3 (14%) and a normal bowel gas pattern in 1 (5%). In all 15 patients with apparent distal colonic obstruction, there was minimal or no gas in the rectosigmoid, with an associated pelvic mass representing the enlarged post partum uterus, which compressed the rectosigmoid and prevented it from filling with gas. All 21 patients had rapid clinical or radiographic improvement on conservative management, indicating a transient post-operative ileus. Radiologists should be aware of the limitations of abdominal plain radiographs following Caesarean section so that a post-operative ileus is not mistaken for a distal colonic obstruction and conservative measures can be undertaken to decompress the bowel until the ileus resolves. PMID- 11064648 TI - Impact of motion artefact on the measurement of coronary calcium score. AB - Reports have shown that total coronary calcium (CC) measured by electron beam CT (EBCT) correlates well with atheroma extent, and this in turn is a reliable indicator of risk for future ischaemic events. Although total CC may be measured using a conventional CT scanner, image quality is degraded by cardiac motion artefact. Errors in CC measurement owing to slice misregistration between adjacent breath-holds affect both conventional CT and EBCT. The latest generation conventional CT scanners have acquisition times of 500 ms or shorter, and, when combined with ECG triggering, quantitative CC measurement without reliance upon EBCT becomes a real possibility. We investigated the effect of motion on the measured calcium score using a moving phantom. Our results show that use of ECG triggering with conventional CT improves reproducibility of Agatston calcium score measurement. Increasing motion time during image acquisition results in an apparent increase in the Agatston CC score. Alternative measures of the amount of CC may be less susceptible to motion-induced bias, but have a similar reproducibility. PMID- 11064649 TI - Digital selenium radiography: anti-scatter grid for chest radiography in a clinical study. AB - The purpose of this study was to evaluate the diagnostic performance of an additional stationary anti-scatter grid in digital selenium radiography (DSR) compared with images acquired with only an air gap. Chest radiographs were obtained with DSR in 100 patients with and without an anti-scatter grid. Four observers scored 12 anatomical landmarks, catheters and wire cerclages for their visualization in both subsets of images. Statistical analysis was performed using a paired t-test. Anatomical landmarks, catheters and wire cerclages were statistically better visualized in regions of high attenuation when the images were performed with an anti-scatter grid. No statistically significant difference was noted for peripheral regions, nor for sex and weight of the patient between the two modalities. Therefore, an anti-scatter grid is not recommended for chest radiography as it increases the radiation exposure of patients without having a significant impact on visualization for all regions of the chest. PMID- 11064650 TI - Hip axis length in an Italian osteoporotic population. AB - Models proposed so far for the pathogenesis of osteoporosis often do not take into account the factors underlying the different incidences of hip fracture in different populations. To address this issue, we identified 34 female patients with hip fracture (HF) and 16 women with at least four vertebral fractures (VF) in a population-based retrospective study. Each participant had a bone mineral density (BMD) measurement of the lumbar spine and hip using a Hologic QDR-2000 scanner, in single beam mode for the latter site. Hip axis length (HAL) was determined automatically (precision 1.5%). HAL derived from 149 normal subjects (age range 19-75 years) was 10.3 +/- 0.5 cm. BMD values found at the femoral neck were almost similar, but differed significantly at the spine between the two groups. Mean values of femur HAL in HF patients (10.55 +/- 0.5 cm) were significantly higher compared with VF patients (9.85 +/- 0.54 cm; p < 0.001). Interestingly, both mean values differed significantly from the mean for normal subjects. Our results demonstrate that patients with multiple vertebral fractures have significantly lower vertebral BMD values but similar femoral neck values compared with patients who fracture at the hip; furthermore, hip axis length was more than 1 SD higher in these latter patients compared with that of VF patients. These results suggest that the size and shape of the hip can explain part of the observed aetiologic differences between these two types of osteoporotic fractures. PMID- 11064651 TI - How should breathing motion be combined with other errors when drawing margins around clinical target volumes? AB - Geometric uncertainties in radiotherapy treatment may be accommodated by drawing an adequate margin around the clinical target volume (CTV). The width of the margin is commonly based upon the quadrature sum of the standard deviations of the contributory errors. This approach is satisfactory when the probability distributions of the component errors are Gaussian, but breathing-induced motion of the CTV is generally not Gaussian. This paper shows that the blurred dose distribution D(z) arising from a step function beam edge that is moving cyclically along the z-axis is given by D(z) = 2t/tau, where t(z) relates time to position and tau is the cycle duration. Applying this relation to realistic breathing models results in a dose distribution in which the sharp edge of the step function is still evident even after blurring caused by the motion. This suggests that breathing-induced motion should be accounted for separately, with the breathing margin added linearly to the quadrature sum of the other contributing errors. PMID- 11064652 TI - Change in oxygenation status in intratumour total and quiescent cells following gamma-ray irradiation, tirapazamine administration, cisplatin injection and bleomycin treatment. AB - C3H/He mice bearing SCC VII tumours received 5-bromo-2'-deoxyuridine (BrdU) continuously for 5 days via implanted mini-osmotic pumps to label all proliferating (P) cells. The mice then received gamma-ray irradiation, or administration of tirapazamine (TPZ), cisplatin or bleomycin. At various time points after each treatment, tumour-bearing mice were irradiated with a series of test doses of gamma-rays, while alive or after being killed, to obtain hypoxic fractions (HFs) in the tumours. Immediately after gamma-ray test irradiation, the tumours were excised, minced and trypsinized. Tumour cell suspensions obtained were incubated with cytochalasin-B, a cytokinesis blocker, and the micronucleus (MN) frequency in cells without BrdU labelling (i.e. quiescent (Q) cells) was determined using immunofluorescence staining for BrdU. MN frequency in the total (P + Q) tumour cells was determined from the tumours that were not pre-treated with BrdU. MN frequency of BrdU-unlabelled cells was then used to calculate the surviving fraction of the unlabelled cells from the regression line for the relationship between the MN frequency and the surviving fraction of total tumour cells. TPZ and cisplatin reduced the HF after treatment, especially in Q cells, and this tendency was particularly marked with TPZ. In contrast, bleomycin increased the HF after treatment. Both reoxygenation following gamma-ray irradiation or bleomycin treatment and a subsequent return to pre-treatment levels of HF following TPZ or cisplatin treatment (rehypoxiation) occurred more rapidly in total (P + Q) cells than in Q cells. Based on our previous report that total (P + Q) and Q cells within this tumour have large acutely and chronically HFs, respectively, we conclude that acute hypoxic cells play a major role in reoxygenation and rehypoxiation in SCC VII tumours. PMID- 11064653 TI - An audit of outcome of adjuvant post-operative radiotherapy for 52 women with stage II carcinoma of the endometrium. AB - A retrospective review was undertaken of the medical records of 52 women with stage II carcinoma of the endometrium who received adjuvant radiotherapy following surgery. The information was obtained from medical notes and a hospital database. Actuarial disease-free survival was 68% at 5 years for those women with stage IIA disease, and 70% at 5 years for those women with stage IIB disease. 6 of the women (11.5%) had side effects from treatment. In contrast to the literature, the only statistically significant prognostic factor in this study was histological differentiation; patients with poorly differentiated tumours fared worse (p = 0.05). This may indicate that a greater number than 52 women is needed to demonstrate weaker prognostic factors such as substage. A larger review is being undertaken of the remaining women recorded on the database, with stage I, III and IV disease. PMID- 11064654 TI - GSM cell phones can interfere with ionizing radiation dose monitoring equipment. AB - Cell phone use is growing worldwide. These phones transmit to adjacent base stations using radiofrequency signals in the microwave range (approximately 900 approximately 1800 MHz). Portable electronic dose monitoring equipment is used in hospitals and other institutions to monitor and control levels of exposure to ionizing radiation, and to reassure staff. The objective of this paper is to investigate the effect of mobile phones on a sample of dose monitoring devices. Two mobile phones (Siemens C25 and Motorola CD930) were used in the study. Field strengths were measured to be in the range 0 V m-1 to over 100 V m-1, depending on the distance from the phone, and were strongest at the beginning of a call. Personal electronic dosemeters (n = 7), portable dose monitors (n = 4) and contamination monitors (n = 2) were assessed. All the units were in service. Three of the personal dosemeters showed abnormal responses when exposed to mobile phone transmission. One dosemeter (Siemens EPD-2) registered doses equivalent to a dose rate of 99 mSv h-1. In addition, two of the portable dosemeters and one of the contamination monitors also gave an abnormal response. Interference was observed across a number of detector types from a number of manufacturers. Modern cell phones can interfere with ionizing radiation dose monitoring equipment. This should be taken into account when distributing these devices and when assessing results generated by them. Electromagnetic compatibility testing should form part of the commissioning and specification protocol for new dose monitoring equipment. PMID- 11064655 TI - Communication of doubt and certainty in radiological reports. AB - We have investigated the reliability of communication of uncertainty in radiological reports. The 18 most commonly used verbal expressions of probability were identified from a series of radiological reports. 11 clinicians (three radiologists, three rheumatologists and five orthopaedic surgeons) recorded the probability that they ascribed to each of the 18 expressions using visual analogue scales. Each subject was re-tested on four occasions at least 1 week apart. The results were analysed to assess reproducibility within and between individuals. We found considerable variation in the probabilities assigned to many commonly used expressions between subjects, and between repeated testing of the same subject. Some expressions were rated much more consistently than others. "Absent", "excludes", "unlikely", "probable", "certain" and "definite" were the most consistently rated expressions. We have identified a potential source of misunderstanding in radiological reports owing to differences in interpretation of expressions used by radiologists and referring clinicians. PMID- 11064656 TI - Measurement of dose in panoramic dental radiology. AB - The National Radiological Protection Board (NRPB) has recommended the introduction of dose-width product (DWP) for the measurement of patient dose in panoramic dental radiology and has proposed a reference level of 65 mGy mm for adult exposures. This paper describes a method for measuring DWP and dose-area product (DAP) using thermoluminescent dosemeters (TLDs). The technique was used on 16 sets with a range of exposure settings. The mean value of DWP was 14% higher than the mean value reported from a survey by the NRPB. This difference is most likely to be caused by systematic variations due to measurement method. The average DAP for a standard adult examination was shown to be 11.3 cGy cm2. Data are presented so that the DAP can be derived from the exposure factors (tube current and operating potential) and beam area. Based on published data for effective dose, it is estimated that the DAP to effective dose conversion factor is approximately 0.06 mSv(Gy cm2)-1. The average DAP value (11.3 cGy cm2) can be compared with the average value for intraoral radiography (9.3 cGy cm2) based on the NRPB survey of entrance surface doses assuming 6 cm circular collimation. PMID- 11064657 TI - Directionality of extruded lithium fluoride thermoluminescent dosemeters in a cobalt-60 beam. AB - An experimental investigation of the directionality of commercially available extruded lithium fluoride (LiF:Mg,Ti) thermoluminescent dosemeters was carried out in a cobalt-60 beam at a water depth of 5 cm. One-half of a batch of 60 chips (3.1 x 3.1 x 0.9 mm3) was exposed face-on (faces perpendicular to the beam central axis), and the other half was exposed in an edge-on orientation (two edges perpendicular and faces parallel to the beam axis). Measurements show that an edge-on exposure results in a thermoluminescence reading approximately 0.9% lower than a face-on exposure. Although this is of minor importance in every day patient dosimetry, it is relevant in evaluating errors in in-phantom dosimetric measurements where greater accuracy is required. PMID- 11064658 TI - Malignant stromal tumour of the rectum: findings at endorectal ultrasound and MRI. AB - This case report describes the findings on endorectal ultrasound and MRI in a patient with a giant malignant stromal tumour of the rectum. A review of imaging characteristics and histopathological findings as described in the literature is presented. PMID- 11064659 TI - CT diagnosis of acute appendicitis in a femoral hernia. AB - We present the first case, to our knowledge, of a patient with a CT diagnosis of acute appendicitis strangulated in a femoral hernia, a known but very rare entity. CT features of acute appendicitis within the hernia established the correct diagnosis. The pathological findings confirmed the diagnosis of this rare location of appendicitis. PMID- 11064660 TI - Recurrent superior vena caval obstruction due to invasion by malignant thymoma: treatment using a stent-graft. AB - A stent-graft was used to palliate superior vena caval obstruction in a 50-year old patient with histologically proven ingrowth of malignant thymoma through three previously inserted non-covered stents. The stent-graft is still patent 12 months later. This is the first report of such a procedure where histological evidence of tumour ingrowth is available and long-term patency is verified. PMID- 11064661 TI - Extensive soft tissue uptake of 99Tcm methylene diphosphonate in a patient with multiple myeloma. AB - Bone scintigraphy is not usually performed in multiple myeloma (MM), as marrow deposits characteristically show no tracer uptake. However, metastatic bone disease often mimics MM both clinically and biochemically, resulting in a substantial number of MM patients undergoing bone scintigraphy. Variable appearances in these cases have been reported, ranging from normal to a superscan, the latter a result of massive tracer uptake within bone. Soft tissue uptake has been documented, often when MM is complicated by secondary amyloidosis. This usually results in mainly solid organ uptake of tracer. We report a case of MM where massive soft tissue uptake occurred, primarily within muscles, with very little isotope elsewhere. PMID- 11064662 TI - Vertigo after stapes surgery: the role of high resolution CT. AB - CT findings in patients with vertigo after stapes surgery include a prosthesis shaft entering the vestibule and compressing the saccule, a complete dislocation of the stapes prosthesis, air bubbles and fluid collections within the vestibule and outside the oval window indicating a perilymphatic fistula, and bony fragments leading to compression of the basal saccule. Although immediate post operative vertigo is often transient, patients with persistent or recurrent vertigo should be imaged as high resolution CT will determine the underlying cause in the majority of cases. PMID- 11064663 TI - Case of the month. A twist in the tail. PMID- 11064664 TI - Modern laboratory diagnosis of mycobacterial infections. AB - This review summarises recent advances made in microscopic techniques (fluorescence and peptide nucleic acids) and culture techniques (solid, liquid, radiometric, and non-radiometric systems) and in the development of rapid methods for the identification of mycobacterial cultures (high performance liquid chromatography, thin layer chromatography, RNA sequencing, and polymerase chain reaction restriction enzyme assays). The role of molecular amplification systems in identifying Mycobacterium tuberculosis is described. Most methods record high specificity and sensitivity for smear positive sputum but have variable sensitivity for sputum smear negative and extrapulmonary specimens. Specimen quality will affect the performance of these assays and organisational delays, such as the batching of specimens, can reduce the time saved. In house assays can be as effective as commercial systems as long as appropriate controls are used. PMID- 11064665 TI - Axillary staging of breast cancer and the sentinel node. AB - Pathological aspects of axillary nodal staging of breast cancer and in particular sentinel lymph node (SLN) biopsy are reviewed. SLN biopsy seems an almost ideal staging procedure because it has both high accuracy and a low false negative rate. It may also allow a cost effective use of more sensitive methods of metastasis detection. However, the biological relevance of metastases detected only by modern tools remains to be elucidated. This review focuses on standard axillary staging and the histopathological investigation of SLNs, with emphasis on the intraoperative setting. Future trends including ancillary studies, quality control issues, prediction of non-SLN involvement, and suggestions concerning the minimum requirements for the histology of axillary SLNs are also discussed. PMID- 11064666 TI - Mammary and extramammary Paget's disease. AB - Mammary and extramammary Paget's disease are uncommon intraepithelial adenocarcinomas. Both conditions have similar clinical features, which mimic inflammatory and infective diseases. Histological diagnostic confusion can arise between Paget's disease and other neoplastic conditions affecting the skin, with the most common differential diagnoses being malignant melanoma and atypical squamous disease. The glandular differentiation of both mammary Paget's disease and extramammary Paget's disease is indicated by morphological appearances, the presence of intracellular mucin in many cases, and positive immunohistochemical staining for glandular cytokeratins, epithelial membrane antigen, and carcinoembryonic antigen. This article provides an overview of mammary and extramammary Paget's disease and discusses recent evidence regarding the cell of origin. The concepts of primary and secondary Paget's disease are presented and the differential diagnosis is discussed with reference to immunohistochemical markers that might be of diagnostic value. PMID- 11064667 TI - Whipple's disease revisited. AB - Whipple's disease has traditionally been considered to be a rare multisystem disorder dominated by malabsorption. The recent identification of the Whipple's disease bacillus has, using polymerase chain reaction based assays, fueled advances in the investigation, diagnosis, and management of this disease. This leader reviews the aetiology, clinical manifestations, investigation, and treatment of Whipple's disease in the light of this new information. PMID- 11064668 TI - Histological identification of Helicobacter pylori: comparison of staining methods. AB - AIM: To determine whether two recently described staining methods (the modified McMullen's and the Helicobacter pylori silver stain HpSS methods) used for the histological identification of H pylori organisms are superior to two established techniques (the modified Giemsa and anti-H pylori antibody immunostain) in terms of availability, reproducibility, rapidity, sensitivity, and cost. METHODS: Histological sections from 63 paired gastric biopsies from adult patients previously investigated for dyspepsia were stained with the four methods and these were assessed blindly and independently by two observers. Of the 63 patients, 30 were originally negative in all tests for H pylori infection, 30 were positive, and the remaining three cases had discordant results using a combination of five tests (rapid biopsy urease test, urea breath test, culture, serology, and histology). RESULTS: Interobserver agreement was best with the antibody method (98%), followed by the McMullen's (90%), Giemsa (87%), and HpSS (85%). Of the 60 "gold standard" positive and negative cases, 30 were positive by the modified Giemsa stain, 29 by the McMullen's method, 29 by HpSS, and 30 by the antibody stain. However, there were two false positives with the HpSS method. The modified Giemsa is the cheapest and easiest to perform technically. CONCLUSIONS: When H pylori are present, careful examination will almost always reveal them, whichever of these stains is used. However, the modified Giemsa stain is the method of choice because it is sensitive, cheap, easy to perform, and reproducible. PMID- 11064669 TI - Routine DNA cytometry of benign and malignant pleural effusions by means of the remote quantitation server Euroquant: a prospective study. AB - AIM: To analyse the practicability and potential assistance of static DNA cytometry performed by means of the remote quantitation server Euroquant and the internet in routine diagnostic analysis of pleural effusions, and to outline the role of DNA cytometry on pleural effusions in distinguishing between benign and malignant (and herein primary versus metastatic) effusions. MATERIALS AND METHODS: Cytological smears of 294 pleural effusions were stained with the Feulgen method. The DNA content of a minimum of 300 randomly chosen analysis nuclei and 30 reference nuclei (lymphocytes) was measured by internet connection to the remote quantitation server Euroquant. Cytometric features were derived from the histograms, and the time needed for case evaluation, the reliability of staining and measurement procedures, and the contribution to the final diagnosis were evaluated. RESULTS: Only 120 of 294 pleural effusions could be measured. The total measurement time for each specimen was 60 minutes. The guidelines of the consensus report on DNA measurements were fulfilled. Seventy eight malignant (18 mesotheliomas, 60 metastatic tumours) and 42 benign effusions were measured. Seven of 78 malignant effusions were euploid and none of 42 benign effusions were aneuploid. The sensitivity and specificity were 91% and 100%, respectively, for distinguishing benign from malignant effusions, and 95% and 100%, respectively, for discriminating between benign and malignant effusions caused by metastatic malignant tumours. CONCLUSIONS: Static DNA cytometry using the remote quantitation server Euroquant can be performed reliably in the routine diagnosis of pleural effusions; however, only 40% of effusions meet the technical requirements for static DNA cytometry. Within the measurable cases, static DNA cytometry made an important contribution to the confirmation/exclusion of malignancy. PMID- 11064670 TI - Qualitative and quantitative differences between bile ducts in chronic hepatitis and in primary biliary cirrhosis. AB - AIM: Lymphocytic infiltration in the portal triads usually conceals the detection -in haematoxylin and eosin (H&E) stained sections--of bile ducts in two liver diseases: chronic hepatitis and primary biliary cirrhosis. The aim was to assess the number and the characteristics of the bile ducts in those diseases with the aid of an antibody to cytokeratin 7 (CK7). METHODS: Consecutive sections from 99 liver biopsies were stained with H&E and anti-CK7. RESULTS: In H&E sections the total number of central bile ducts in the triads was 52 in primary biliary cirrhosis (n = 37), 69 in chronic hepatitis (n = 43), and 30 in miscellaneous cases (n = 19). Using anti-CK7, the number of central bile ducts was 276 in primary biliary cirrhosis, 348 in chronic hepatitis, and 96 in miscellaneous cases. Central bile ducts with lumen were found in 93.0% of chronic hepatitis cases and in 89.5% of the miscellaneous cases, but in only 13.5% of the primary biliary cirrhosis cases. Peripheral bile ducts in groups of > or = 4/triad were found in all cases of chronic hepatitis (100%) and in 75.7% primary biliary cirrhosis cases, but only in 10.5% of the miscellaneous cases. In 21.6% of primary biliary cirrhosis cases, no bile ducts (central and/or peripheral) were present. CONCLUSIONS: Anti-CK7 detects bile ducts in the triads that are concealed by chronic inflammatory cells. Central and peripheral bile ducts in groups of > or = 4 were significantly more common in primary biliary cirrhosis and chronic hepatitis than in other liver diseases. The lack of lumen in central bile ducts, as well as the absence of central and/or peripheral bile ducts in CK7 stained liver sections, seem to be valuable additional parameters in the differential diagnosis between primary biliary cirrhosis and chronic hepatitis. PMID- 11064671 TI - Expression of human alpha-defensin 5 (HD5) mRNA in nasal and bronchial epithelial cells. AB - BACKGROUND/AIMS: Human defensins are antibiotic peptides expressed in myeloid and epithelial cells. Human alpha-defensin 5 (HD5) has been detected in Paneth cell granules in the crypts of Lieberkuhn and has recently been identified in the female reproductive tract. The aim of this study was to investigate the presence of HD5 mRNA in nasal and bronchial epithelial cells. METHODS/RESULTS: Semiquantitative reverse transcription polymerase chain reaction (RT-PCR) analysis showed that HD5 mRNA was expressed infrequently and to varying degrees in bronchial and nasal epithelial cells. In situ hybridisation resulted in a positive signal in the epithelial layer of nasal polyps. HD5 mRNA was locally restricted to a specific area of epithelial cells and also occurred in submucosal glands. CONCLUSIONS: HD5 mRNA expression in nasal and bronchial epithelial cells is rare and seemed to be locally induced. The results indicate that HD5 might play a role in innate defence in nasal and bronchial epithelia. PMID- 11064672 TI - Indirect immunofluorescence (IIF) of normal washed peripheral blood cells to demonstrate antineutrophil cytoplasmic antibodies (ANCA). AB - BACKGROUND: The "International consensus document on testing and reporting of antineutrophil cytoplasmic antibodies (ANCA)" requires all sera to be examined by indirect immunofluorescence (IIF). However, commercial neutrophil slides are expensive, fluorescence patterns can be difficult to interpret, and coincidental antinuclear antibodies (ANA) cannot be demonstrated; in addition, in house cytospin neutrophil preparations are time consuming to prepare and deteriorate with time. AIMS: To compare the IIF demonstration of ANCA, using washed peripheral blood cell smears, with commercial neutrophil preparations and with ANCA positivity as demonstrated by enzyme linked immunosorbent assay (ELISA). METHODS: Serum fluorescence positivity, pattern, and intensity using washed peripheral blood cell smears were compared with the results obtained using commercial neutrophil slides (INOVA). Fluorescence positivity, pattern, and intensity of 500 sera from consecutive patients with suspected vasculitis tested with washed peripheral blood cells were compared with binding in ELISAs for proteinase 3 (PR3) and myeloperoxidase (MPO). RESULTS: IIF of washed peripheral blood cell smears detected seven of eight sera with cytoplasmic fluorescence (C ANCA), and 11 of 12 sera with perinuclear fluorescence (P-ANCA) demonstrated using commercial slides. The two sera that were negative by IIF were also negative in the ELISAs for both PR3-ANCA and MPO-ANCA. Of the 500 sera examined, there were 35 (7%) with C-ANCA, 65 (13%) with P-ANCA, and eight (2%) IIF negative sera that were positive by either ELISA. There was a strong correlation between C ANCA fluorescence and PR3-ANCA values (p < 0.0001), and a moderate to strong correlation between P-ANCA fluorescence and MPO-ANCA values (p < 0.001) when ANCA fluorescence was demonstrated with washed peripheral blood cell smears. CONCLUSIONS: Washed peripheral blood cells are a convenient and useful low cost alternative to commercial or cytospin neutrophil preparations for the IIF demonstration of ANCA. PMID- 11064673 TI - Abnormal regulation of the oestrogen receptor in benign breast lesions. AB - BACKGROUND: In normal breast tissue the oestrogen receptor (ER) and the proliferation associated antigen Ki67 are negatively associated, indicating that ER+ cells are non-dividing, or that the receptor is downregulated as cells enter cycle. This relation is completely or partially lost in many ER+ breast cancers and in in situ proliferations associated with an increased cancer risk, where coexpression of the two markers is often found. AIMS: To determine whether similar changes can be identified in other risk associated breast lesions. PATIENTS/METHODS: Paraffin wax blocks from 12 cases of lactational change, 21 apocrine metaplasias, 22 duct ectasias, 20 sclerosing adenosis, 20 fibroadenomas, 19 phyllodes tumours, 20 radial scars, 21 papillomas (15 solitary and six multiple), 15 gynaecomastias, and nine postmortem male breast tissues were retrieved. Immunohistochemistry was used to determine the expression of ER and dual labelling immunofluorescence was used to detect cells expressing both ER and Ki67. RESULTS: Increased numbers of ER+ cells were seen in sclerosing adenosis, radial scars, papillomas, fibroadenomas, and phyllodes tumours but not in apocrine cysts (where no ER+ cells were detected) or duct ectasia (where normal numbers were found). As in the normal breast, the proportion of ER+ cells increased with age in all lesions with the exception of fibroadenomas. Coexpression of ER and Ki67 was found in an increased proportion of cells of all risk associated lesions studied. ER+ cells were less likely to be dividing than ER- cells in all cases, although this was significant only for sclerosing adenosis. The data on sclerosing adenosis, radial scars, papillomas, and fibroadenomas are comparable with those reported previously in hyperplasia of usual type, whereas those in duct ectasia are similar to those of the normal breast. The findings in all lesions, however, differed from those in ductal carcinoma in situ, where proportions of ER+ and ER+/Ki67+ cells are higher and the relation between ER+ cell numbers and age is lost. Thus, the nature and degree of dysregulation of ER in benign breast lesions is broadly in accordance with the degree of risk of developing breast cancer with which they are associated. In gynaecomastia, the proportions of ER+ and ER+/Ki67+ cells were comparable with those seen in benign female breast lesions, but changes with age were not observed. However, the changes in gynaecomastia were similar to those seen in normal male breast. CONCLUSION: These findings are in keeping with the contention that the dissociation of ER and Ki67 expression is a very early change in the pathway to many breast cancers. However, this change might only have preneoplastic importance in the hormonal milieu of the female breast. PMID- 11064674 TI - Cell proliferation in type C gastritis affecting the intact stomach. AB - AIMS: Type C gastritis caused by bile reflux has a characteristic appearance, similar to that seen in other forms of chemical gastritis, such as those associated with NSAIDs or alcohol. An increase in mucosal cell proliferation increases the likelihood of a neoplastic clone of epithelial cells emerging, particularly where there is chronic epithelial injury associated with bile reflux. It has been shown previously that type C gastritis is associated with increased cell proliferation in the postsurgical stomach. The aim of this study was to determine cell proliferation in type C gastritis caused by bile reflux affecting the intact stomach. METHODS: Specimens from 15 patients with a histological diagnosis of type C gastritis on antral biopsy were obtained from the pathology archives between 1994 and 1997. A control group of nine normal antral biopsies was also selected and all underwent MIB-1 immunostaining. The gastric glands were divided into three zones (zone 1, gastric pit; zone 2, isthmus; and zone 3, gland base) and the numbers of positively staining nuclei for 500 epithelial cell nuclei were counted in each zone to determine the percentage labelling index (LI%). RESULTS: Cell proliferation was significantly higher in all three zones of the gastric glands with type C gastritis compared with controls as follows: zone 1, median LI% in type C gastritis 64.7 (range, 7.8 99.2), controls 4.7 (range, 2.0-11.3); zone 2, median LI% in type C gastritis 94.7 (range, 28.8-98.7), controls 40.2 (range, 23.1-70.3); and zone 3, median LI% in type C gastritis 20.0 (range, 1.3-96.0), controls 2.6 (range, 0.9-8.7). CONCLUSIONS: Bile reflux is thought to act as a promoter of gastric carcinogenesis in the postsurgical stomach. The same may be true in the intact stomach. PMID- 11064675 TI - Histiocytic lymphoma presenting as a testicular tumour and terminating in acute monoblastic leukaemia. AB - A 58 year old man presented in 1995 with a swollen testicle. After orchidectomy, a diagnosis of poorly differentiated lymphoma was made. Lymphoid, epithelial, and seminoma markers were all negative. Six months later he developed a buccal lesion, which was biopsied and reported as a high grade non-Hodgkin's lymphoma. It responded completely to chemotherapy but within a year he developed a forearm swelling, which was biopsied and imprints made before fixation of the material. Immunocytochemistry on the imprints showed positivity with antibodies to CD4, CD68, and muramidase, and the non-specific esterase cytochemical stain was strongly positive, leading to a diagnosis of true histiocytic lymphoma. Despite further treatment, the patient entered a terminal acute leukaemic phase, the blasts marking as monoblasts. Review of all the biopsies, including molecular investigations and further immunohistochemistry studies performed retrospectively on the original biopsy, demonstrated that this was the same malignant cell line throughout, and we conclude that this is a case of histiocytic lymphoma, initially presenting as a testicular tumour and terminating in acute monoblastic leukaemia. A diagnosis of histiocytic lymphoma should be considered when lymphoid markers are negative in an apparent lymphoma, but should not be made without recourse to appropriate immunophenotypic and molecular studies. PMID- 11064676 TI - An investigation of the Peutz-Jeghers gene (LKB1) in sporadic breast and colon cancers. AB - AIMS: To explore the role of the Peutz-Jeghers gene (LKB1) in sporadic breast and colon cancers. METHODS: Thirty consecutive sporadic carcinomas of the breast and 23 of the colon were selected. DNA was extracted from paraffin wax embedded tissue and analysed for loss of heterozygosity (LOH) at microsatellite markers D19S886 and D19S565 close to the LKB1 gene. Tumours showing LOH were screened for LKB1 mutations by single strand conformational polymorphism (SSCP). RESULTS: Five breast carcinomas showed LOH (21% and 7% of those informative for D19S886 and D19S565, respectively). Five of the colorectal carcinomas showed LOH (15% and 36% of those informative for D19S886 and D19S565, respectively), with one sample showing allele loss with both markers. Screening of these 10 carcinomas by SSCP identified one migrational shift but sequencing revealed an intronic polymorphism only. Therefore, no coding mutations were found in these carcinomas. CONCLUSIONS: These findings suggest that although allele loss at the LKB1 locus occurs relatively frequently in sporadic breast and colon cancers, mutations do not seem to be a feature. PMID- 11064677 TI - Fatal methane and cyanide poisoning as a result of handling industrial fish: a case report and review of the literature. AB - The potential health hazards of handling industrial fish are well documented. Wet fish in storage consume oxygen and produce poisonous gases as they spoil. In addition to oxygen depletion, various noxious agents have been demonstrated in association with spoilage including carbon dioxide, sulphur dioxide, and ammonia. A fatal case of methane and cyanide poisoning among a group of deep sea trawler men is described. Subsequent independent investigation as a result of this case led to the discovery of cyanides as a further potential noxious agent. This is thus the first case in which cyanide poisoning has been recognised as a potentially fatal complication of handling spoiled fish. The previous literature is reviewed and the implications of the current case are discussed. PMID- 11064678 TI - A paradoxical severe decrease in serum HDL-cholesterol after treatment with a fibrate. AB - There have been a handful of reports in the literature of a paradoxical decrease in serum high density lipoprotein (HDL)-cholesterol in patients on fibrate drugs. The reason for this decline in cardioprotective HDL-cholesterol is not known and may have potential deleterious effects on the patient. This report describes a decrease in serum HDL-cholesterol in a patient on both simvastatin and bezafibrate. This patient also developed abnormal renal function, probably interstitial nephritis. In addition, the literature of fibrate induced serum HDL cholesterol decline is reviewed and possible mechanisms for this phenomenon discussed. PMID- 11064679 TI - Multifocal aggressive angiomyxoma: a case report. AB - A case of aggressive angiomyxoma in a 25 year old woman is presented. The patient was admitted to hospital with a history of hesitancy of micturation and pain in the right iliac fossa. She was found to have a left labial mass, which was clinically diagnosed to be a Bartholin gland cyst. A pelvic ultrasound revealed an additional mass in the right paravesical region. At surgery, two distinct masses were removed, one from the right perivesical space and the other from the left labium. Both masses were rubbery, white, and gelatinous and showed similar histopathology findings of thick and thin walled vascular channels set in a loose myxoid stroma. A diagnosis of multifocal aggressive angiomyxoma was made. This is the first reported case of aggressive angiomyxoma occurring as two distinct masses in one patient. PMID- 11064680 TI - Coexistence of an endocrine tumour in a serous cystadenoma (microcystic adenoma) of the pancreas, an unusual association. AB - A pancreatic endocrine tumour arising within a serous cystadenoma is reported. A 49 year old woman was admitted with a history of epigastric pain, nausea, vomiting, and weight loss of two months duration. She had been diabetic for 12 years. An epigastric mass was palpated in the physical examination, and computed tomography revealed a multiloculated cystic lesion in the pancreas. Pathological examination of the pancreatic tumour revealed the coexistence of a serous cystadenoma and an endocrine tumour. The endocrine tumour, which was located inside the serous cystadenoma, was 1 cm in diameter. The first case of a serous cystadenoma of the pancreas containing a pancreatic endocrine tumour was reported in the literature recently. This paper reports another incidentally found pancreatic endocrine tumour arising within a serous cystadenoma. PMID- 11064681 TI - Angiomyofibroblastoma of the vagina. PMID- 11064682 TI - Thrombophilia testing. PMID- 11064683 TI - A comparison of international normalised ratio (INR) measurement in hospital and general practice settings: evidence for lack of standardisation. PMID- 11064684 TI - On surviving cancer. PMID- 11064685 TI - Hospitalists for the NHS? PMID- 11064686 TI - Sonoclot coagulation analysis of in-vitro haemodilution with resuscitation solutions. AB - The colloid and crystalloid solutions used for resuscitation should preferably be free from effects on coagulation. In 10 volunteers, the effects of haemodilution with various concentrations of 0.9% sodium chloride and 4% succinylated gelatin were assessed by Sonoclot analysis, which describes the whole coagulation process. Small and moderate haemodilution (up to 40%) with 0.9% sodium chloride promoted coagulation. Similar haemodilution with 4% succinylated gelatin impaired coagulation, and at 60% haemodilution coagulation was very poor. These findings need to be confirmed in vivo and their clinical relevance determined. PMID- 11064687 TI - Costs of seeking ethics approval before and after the introduction of multicentre research ethics committees. AB - With the advent of multicentre research ethics committees in the UK, local research ethics committees (LRECs) are required to advise only on issues relating to the local acceptability of a project. We looked at the handling of two commercially sponsored studies, one initiated before the change and one after, confining the analysis to 21 LRECs approached in both. As judged by the amount of paper per application, the new system for LRECs is simpler and should be less costly. However, there was an increasing tendency for LRECs to charge for their services (30% study 1, 47% study 2) and these charges varied by more than 400%. If such fees must be levied, a common scale is desirable. PMID- 11064688 TI - Xenon anaesthesia. PMID- 11064689 TI - Cancer chemoprevention: a clinical reality. PMID- 11064690 TI - Early identification of autism by the CHecklist for Autism in Toddlers (CHAT). PMID- 11064691 TI - Whiplash injury: why are we achieving so little? PMID- 11064692 TI - Knowledge for the clinician. 7. Intranets. PMID- 11064693 TI - Headache, hypertension and Horner's syndrome. PMID- 11064694 TI - Surgical management of Mycobacterium avium intracellulare infection in children. PMID- 11064695 TI - Tuberous sclerosis--could it be? PMID- 11064696 TI - Minimal change nephropathy with adenocarcinoma of breast. PMID- 11064697 TI - Plague in east Suffolk 1906-1918. PMID- 11064698 TI - Air pollution and death. PMID- 11064699 TI - The emperor with the shaking head. PMID- 11064700 TI - Drawings of changes in scrofulous glands by Frederick Treves. PMID- 11064701 TI - Continuing legacy of the Third Reich. PMID- 11064702 TI - Experimental allergic encephalomyelitis and multiple sclerosis. PMID- 11064703 TI - Cerebral venous thrombosis: the first reported case of adolescent stroke? PMID- 11064704 TI - Plague, rats and the Bible. PMID- 11064705 TI - Albumin revisited. PMID- 11064706 TI - Heparin for colorectal cancer. PMID- 11064707 TI - Tendosynovial sarcoma. A clinicopathologic review of foot cases with a case report. PMID- 11064708 TI - Pigmented skin lesions. PMID- 11064709 TI - Basal cell carcinoma in the lower extremities. PMID- 11064710 TI - A laboratory review of 67,000 foot tumors and lesions. PMID- 11064711 TI - Clear cell sarcoma of tendons and aponeuroses. A review and case report. PMID- 11064712 TI - Epithelioma cuniculatum of the foot. Literature survey and case history. PMID- 11064713 TI - Soft tissue and bone sarcomas. PMID- 11064714 TI - Malignant fibrous histiocytoma, myxoid variant. PMID- 11064715 TI - Chondromyxoid fibroma. A rare benign bone tumor. PMID- 11064716 TI - Granular cell myoblastoma of the foot. A report of four cases. PMID- 11064718 TI - [In Process Citation] AB - The complex study of the indices of local immune and natural resistance in the parietal mucus of the gastroduodenal zone and general immunity in 174 patients with duodenal ulcer disease before and after surgical treatment was performed. Pronounced suppression of local immunity, presence of autoimmune processes having influence on its indices were noted. After the organ-preserving operations with an ulcer excision, the immune indices restored more rapidly than after gastric resection and vagotomy without an ulcer excision. PMID- 11064717 TI - Commencement address. California College of Podiatric Medicine. Sunday, May 20, 1984. PMID- 11064719 TI - [In Process Citation] AB - The results of treatment of 13,246 patients with acute surgical disease of the abdominal organs have been analysed. In 48 patients, bleeding from a stress ulcer occurred. The symptoms of bleeding were studied. The variants of the clinical course of postoperative acute erosion and ulcer, such as the lightning-like, double-phase, single-phase ones, have been distinguished. Their predictive value is shown. In 89.6% of the patients, a postoperative stress ulcer developed in diseases of the hepatopancreatobiliary zone organs. PMID- 11064720 TI - [In Process Citation] AB - Analysis of the results of surgical treatment of 59 patients with gastroduodenal bleeding for 1991, including 41 with a duodenal ulcer, was carried out. When compared to the previous period, the lethality decreased almost 6-fold. This was associated with revision of the so-called active-expectant tactics. Performance of an emergency intervention is justified in persisting bleeding, and is stopped bleeding, but with a real danger of it recommencement within the nearest 3 days. PMID- 11064721 TI - [In Process Citation] AB - A 10-year experience with the use of vagotomy with the organ-preserving intervention in the treatment of patients with a bleeding pyloroduodenal ulcer who were treated for urgent and emergency indications is presented. The postoperative mortality was 8.1%. An excellent and good long-term result was noted in 88% of the patients, an unsatisfactory one--in 12%. PMID- 11064722 TI - [In Process Citation] AB - Twenty eight children were treated in the clinic for acute gastro-esophageal bleeding in portal hypertension syndrome. Conservative therapy has proved to be effective in 85.8% of cases. Endoscopic sclerotherapy was successfully used in 9 of the 11 patients. In ineffective conservative treatment for 24-48 hrs from the onset of bleeding, the authors consider suturing the esophago-gastric junction with ligation of the gastric vein and blocking the arterial splenic blood flow as an operation of choice. PMID- 11064723 TI - [In Process Citation] AB - The results of treatment of 639 patients with acute esophagogastroduodenal bleeding have been analysed. There were the following causes of bleeding: in 66.4%--ulcer disease, in 9.1%--gastric tumours, in 7.3%--erosive gastritis, in 6.4%--portal hypertension, in 9.1%--the other. In 1.7% of patients, a cause was not established. Conservative treatment was performed in 59% of patients, surgery -in 41%. Complications developed in 24.9% of the patients, 8.6% died. PMID- 11064724 TI - [In Process Citation] AB - We performed 58,938 operations on the abdominal organs. Three hundred and six (37.8%) patients developed early postoperative acute ileus. After its operative elimination, 22(7.2%) patients required performance of a reoperation. Ninety (29.4%) patients died. Of them in 72, death was caused by peritonitis, in 18- pneumonia, acute cardio-vascular failure, impairement in cerebral circulation and pulmonary thromboembolism. PMID- 11064725 TI - [In Process Citation] AB - Prolonged decompression of the small intestine was used in the complex surgical treatment of 85 patients with acute ileus (67) and peritonitis (18). The main stage of the operation included liquidation of ileus and elimination of a source of peritonitis. Antegrade nasogastral intubation of the intestine was performed in 54 patients, via gastrostomy--in 3, retrograde intubation via cecostomy--in 15, via appendicostomy--in 4, via ileostomy--in 9. Performance of active transtubal sanation of the intestine by means of antiseptic solutions (53 cases) in combination with enterosorption (17) contributed to essential increase in the effectiveness of treatment. PMID- 11064726 TI - [In Process Citation] AB - Analysis of the observations of 431 patient with acute diffuse peritonitis is presented. Use of intraabdominal flowing lavage is recommended in patients with inflammatory phenomena against the background of moderately pronounced changes in microcirculation in the parietal peritoneum. The maximum clinical effect of hemosorption and lymphosorption was noted at the first 3 days after the operation in cathepsin-D activity in the blood equal to 14-17 activity units/(min.mg) and lymphorrhea over 700 ml/day. Lethality was 18.8%. PMID- 11064727 TI - [In Process Citation] AB - The results of surgical intervention with the use of laparostomy in 29 patients with diffuse purulent peritonitis at the terminal stage are presented. In 11 cases, laparostomy was performed during the first operation, in 18--for postoperative peritonitis. At the time of operations, laser radiation, gastrointestinal intubation were widely used. PMID- 11064728 TI - [In Process Citation] AB - The effect of hemosorption on motor-evacuatory intestinal function was studied in 169 patients with different forms of peritonitis of appendicular genesis. Early inclusion of hemosorption into the complex of treatment contributed to restoration of motor-evacuatory function of the intestine in 2-3 days, as well as to prevention of development of early commissural ileus. PMID- 11064729 TI - [In Process Citation] AB - The results of treatment of 28 patients with "appendicitis-like" form of the Crohn's disease are summarized. Diagnosis before the operation was supposed in 6 patients. Operated on were 25 patients. All of them underwent appendectomy. Differential diagnosis of acute appendicitis and Crohn's disease in difficult, of main importance are the data of clinical investigation. In diagnosis verification by the data of histologic conclusion, the performance of postoperative specific untiinflammatory therapy is mandatory. Danger of appendectomy under such conditions is exaggerated. The result of treatment at the period of from 3 to 20 years was studied in 22 patients operated on. Performance of postoperative treatment contributed to abatement of the disease, achievement of a satisfactory long-term result in all the patients. PMID- 11064730 TI - [In Process Citation] AB - After operative intervention in 61 of the 5704 patients, as necessity in performance of relaparotomy arose. Mortality after its use was 38%. PMID- 11064731 TI - [In Process Citation] AB - The main forms of periodic disease are presented, clinical cases with the different forms are described. A special attention is paid to the problems of diagnosis and treatment in the emergency surgery of the abdominal organs is abdominal form of the disease. PMID- 11064733 TI - [In Process Citation] AB - In the experiment on 35 dogs, it was established that the normal pancreas had constant bioimpedance parameters: ohmic resistance, polarization coefficient and capacitance. In development of pancreonecrosis, the capacitance increases, and the ohmic resistance and polarization coefficient decrease. PMID- 11064732 TI - [In Process Citation] AB - For 10 years, 26 patients underwent treatment for complications caused by presence in the stomach and intestine of the foreign bodies with low radio opacity and echogenicity. All the patients were operated on. At operation, a cause of the disease was established: in 17 cases--a fishbone, in 5--a small chicken bone, in 1--a conifer needle, in 2--a wooden chip, in 1--a home-made toothpick. PMID- 11064734 TI - [In Process Citation] AB - In dogs with experimental peritonitis, accumulation of toxic metabolic products caused impairement in lipid metabolism. With reduction in pronouncement of endotoxemia under the influence of ultra-violet irradiation, the blood level of biologically active substances decreased, the process of lypolysis was suppressed. PMID- 11064735 TI - [In Process Citation] PMID- 11064736 TI - [In Process Citation] PMID- 11064737 TI - [In Process Citation] PMID- 11064738 TI - [In Process Citation] PMID- 11064739 TI - [In Process Citation] PMID- 11064740 TI - [In Process Citation] PMID- 11064741 TI - [In Process Citation] PMID- 11064743 TI - [In Process Citation] PMID- 11064742 TI - [In Process Citation] PMID- 11064744 TI - [In Process Citation] PMID- 11064745 TI - [In Process Citation] PMID- 11064746 TI - [In Process Citation] PMID- 11064748 TI - [In Process Citation] PMID- 11064747 TI - [In Process Citation] PMID- 11064749 TI - [In Process Citation] PMID- 11064750 TI - [In Process Citation] PMID- 11064751 TI - [In Process Citation] PMID- 11064752 TI - [In Process Citation] PMID- 11064753 TI - [In Process Citation] PMID- 11064754 TI - [In Process Citation] PMID- 11064755 TI - Electrolyte derangement in cerebral malaria: a case for a more aggressive approach to the management of hyponatraemia. AB - Although hyponatraemia has been consistently shown to occur in a large proportion of children with cerebral malaria, no statistical relationship has been established between the incidence of hyponatraemia and that of malaria attributable mortality. However, hyponatraemia is not a benign state in other conditions (such as meningitis) or in surgical patients, and is likely to add to malarial deaths. The high mortality rate seen among cases of cerebral malaria, despite all efforts to curb it, therefore calls for a more aggressive approach to the management of hyponatraemia. Current methods for the administration of hypotonic saline and isotonic glucose solutions need review. In addition, children admitted with cerebral malaria should have their electrolyte status monitored to identify new or ongoing hyponatraemia. When hyponatraemia is discovered, it should be quickly and actively corrected. PMID- 11064756 TI - Comparative clinical characteristics and response to oral antimalarial therapy of children with and without Plasmodium falciparum hyperparasitaemia in an endemic area. AB - The clinical characteristics and the responses to oral antimalarial therapy of 104 children presenting consecutively with or without Plasmodium falciparum hyperparasitaemia (HP) were investigated in an endemic area. At presentation, although the 52 children with HP were significantly younger and had significantly higher heart rates than the 52 without, there were no significant differences between the two groups in their symptoms or in any other clinical feature of their malaria. Responses to oral antimalarial drugs were similar in both groups. Analysis of the disposition kinetics of parasitaemia, using a non-compartmental model similar to that used in characterizing drug disposition, showed that the two groups had similar half-lives of parasitaemia (t1/2pd), volumes of blood completely cleared of parasites per unit time (CLBpd), and parasite-clearance time:t1/2pd ratios. Three children in the HP group, all aged < 3 years, progressed to cerebral malaria within 8 h of presentation, and another HP child presented with isolated trunkal ataxia, indicative of cerebellar involvement. No child in the non-HP group had any of the features of severe malaria. Although the clinical characteristics and responses to oral therapy of children with and without HP are therefore very similar, young children with HP appear to have an increased risk of developing other features of severe malaria. PMID- 11064757 TI - The use of PCR in the diagnosis of hyper-reactive malarial splenomegaly (HMS). AB - Between August 1997 and September 1998, 14 cases of hyper-reactive malarial splenomegaly (HMS) were diagnosed in the Instituto de Salud Carlos III in Madrid, Spain. These cases, from Equatorial Guinea and Cameroon, were identified using the diagnostic criteria established by Y. M. Fakunle in 1981: gross splenomegaly; high levels of anti-malarial antibodies; IgM in serum at least two standard deviations above the local mean; and clinical and immunological response to antimalarial treatment. Although malarial parasites were only detected in the Giemsa-stained blood films of four of the cases, these four and four others were found to have the DNA of such parasites in their blood when tested using a method based on a semi-nested, multiplex PCR. These result indicate that malarial parasitaemias may be more prevalent in HMS than is usually recognized. PMID- 11064758 TI - Chloroquine and desethylchloroquine concentrations in blood cells and plasma from Indian patients infected with sensitive or resistant Plasmodium falciparum. AB - The sensitivities of 61 Indian cases of Plasmodium falciparum malaria to chloroquine (CQ) were investigated using in-vitro and in-vivo methods. Concentrations of CQ and desethylchloroquine (DCQ) in blood cells and plasma from CQ-sensitive and -resistant cases were determined 2 and 7 days after initiation of treatment, by HPLC. On day 2, the mean CQ concentrations in the samples collected from the sensitive cases were higher than those in the samples from the resistant patients, in plasma (0.47 v. 0.32 microgram/ml; P < 0.02) and particularly in the blood cells (1.51 v. 0.46 micrograms/ml; P < 0.001). By day 7, however, the CQ concentrations in the two groups were similar. Although, on day 2, the mean ratio of the CQ to DCQ concentrations was significantly higher in the blood cells from the sensitive group than in those from the resistant cases (P < 0.01), the CQ/DCQ ratios for the plasma were similar for the two groups. Similarly, the mean ratio between the blood-cell concentration of CQ on day 2 and the concurrent plasma concentration (BPr) was also relatively high in the sensitive group (P < 0.001). PMID- 11064759 TI - Systematic mapping of hearts from chronic chagasic patients: the association between the occurrence of histopathological lesions and Trypanosoma cruzi antigens. AB - To investigate the role played by Trypanosoma cruzi in the pathogenesis of chronic chagasic cardiopathy, the myocardiums of 12 hearts from individuals who had this condition were mapped in detail. Attempts were made to associate the histopathological lesions observed with the presence of parasitic antigens in the affected tissue. Samples from 26 regions of each heart were submitted to histological analysis and immunostained for the presence of T. cruzi. All cases showed at least one positive region for antigens, but the quantity of antigen detected varied greatly. The regions showing the greatest pathological changes were the inferior atrial septum, the basal and apical portions of the ventricular septum, and the posterior basal and lateral pre-apical regions of the left ventricle. The posterior wall of the right atrium and the posterior basal wall of the left ventricle presented the greatest intensities of inflammation. Fibrosis was intense in the right atrium, the posterior basal and pre-apical left ventricular free walls and the apex. The regions in which T. cruzi antigens were detected showed the most intense inflammation. However, as there was no significant correlation between the intensity of the lesions and the quantity of parasitic antigen, other mechanisms, such as auto-immunity or hypersensitivity, may stimulate the inflammation. PMID- 11064760 TI - The effects of irrigated agriculture on the transmission of urinary schistosomiasis in the Middle and Upper Valleys of the Senegal River basin. AB - The importance of the increase in irrigated land on the perimeters of the Middle and Upper Valleys of the Senegal River basin, on the prevalence and intensity of urinary schistosomiasis, was investigated. Surveys were conducted, in May-June 1997, to determine the prevalence and intensity of Schistosoma haematobium infection among 1445 children aged 7-14 years: 1011 in 10 villages near Matam, and 434 in four villages near Bakel. Macrohaematuria was present in seven of the study villages (four near Matan and three near Bakel), whereas microhaematuria was present in all the villages, with prevalences of 10%-73%. A second survey, conducted, in June 1999, on 755 children from nine of the study villages near Matam, demonstrated significant increases in the prevalences of both micro- and macro-haematuria in three of the villages, all of which were adjacent to the Senegal River and practising irrigated agriculture. None of the other study villages re-surveyed was irrigating any of its agricultural land. A longitudinal survey was also carried out, between May 1997 and November 1998, on about 10% of the population (2272 subjects) of Nguidjilone, north of Matam; selective treatment with praziquantel (40 mg/kg) was given in May 1997, and mass treatment in May 1998. The data analysed were those relating to the 125 individuals who provided samples at each survey. Very severe infections (> 1000 eggs/10 ml urine) were seen in five subjects in May 1997. One year later (i.e. 1 year after the selective treatment), the prevalence of urinary schistosomiasis had increased in every age-group. Although prevalence had decreased slightly by November 1998 (6 months after the mass treatment), the intensity of the infections seen had increased in every age-group. At the end of the dry season (May-June 1997), Bulinus truncalus infected with schistosome cercariae were recovered from the Senegal River. However, immediately after the next rainy season (November 1997), no snails were found at any collection site on the river. PMID- 11064761 TI - Mapping of lymphatic filariasis in India. AB - The derivation of detailed epidemiological maps, at the relevant spatial resolution, is being increasingly recognized as vital to the effective design and implementation of successful programmes for the control of parasites and their vectors. Geographical information systems (GIS) and a recently complied database on the distribution of lymphatic filariasis in India have now been used to develop the first maps at district-level (i.e. the level at which control against this parasite will be enacted in India) of filariasis endemicity in this country. The derived maps indicate both the substantial extent as well as the marked variability in the geographical distribution of this disease in India. The causative infection and/or the symptomatic disease were detected in most (257) of the 289 districts surveyed up to 1995. Currently there may be up to 27.09 million microfilaraemics, 20.83 million cases of symptomatic filariasis, and about 429.32 million individuals potentially at risk of infection in the country. Probability mapping, based on data quantiles, clearly indicates that the risk of filarial infection in India is not constant throughout the country but exhibits strong regional trends. Filariasis in general may be a particular problem of the eastern half of the country. The results indicate the potentially vital role that GIS based mapping approaches can play in the development of filariasis-control campaigns in India and elsewhere. PMID- 11064762 TI - Two-year follow-up of the microfilaraemia of asymptomatic brugian filariasis, after treatment with two, annual, single doses of ivermectin, diethylcarbamazine and albendazole, in various combinations. AB - Repeated, single, oral doses of combinations of ivermectin, diethylcarbamazine (DEC) or albendazole are recognized as important tools for parasite control in lymphatic filariasis. In order to assess the effects of re-treatment using these combinations in Brugia malayi infections, 40 asymptomatic microfilaraemics were re-treated at the end of the first year, with an additional, single, dose of the combination they had previously received. They were then followed-up for another year. The subjects, of both sexes and aged 14-70 years, each received a two-drug combination: ivermectin (200 micrograms/kg) with DEC (6 mg/kg); ivermectin (200 micrograms/kg) with albendazole (400 mg); or DEC (6 mg/kg) with albendazole (400 mg). The kinetics of microfilarial clearance were similar to that seen during the first treatment, the members of the two groups given DEC having less intense microfilaraemias, 1 year after the re-treatment, than those given ivermectin with albendazole (P < 0.001 for each comparison). At this time, the two DEC groups also had a higher proportion of amicrofilaraemic individuals (22 of 26) than the ivermectin + albendazole group (three of nine). There were fewer adverse reactions in all the groups after re-treatment than seen after the first treatment. In countries such as India, where there is no co-endemicity of onchocerciasis or loiasis, the options for control programmes in areas where brugian filariasis is endemic are DEC alone or DEC in combination with ivermectin or albendazole. Where there is no access to ivermectin, transmission control must be based on DEC alone or in combination with albendazole. PMID- 11064763 TI - Persistence of antibodies to the surface antigen of the hepatitis B virus (anti HBs) in children subjected to the Expanded Programme on Immunization (EPI), including hepatitis-B vaccine, in Thailand. AB - Will hepatitis-B vaccine administered at birth, and at 2 and 6 months of age, as an integral part of Thailand's Expanded Programme on Immunization, provide long term protection? In an attempt to answer this question, residents of five provinces (representing five distinct geographical areas of Thailand) who were aged 1-10 years and had received this course of vaccination were enrolled on a serological study. Each was tested, with ELISA, for the surface antigen of hepatitis B (HBsAg) and for antibodies against this antigen (anti-HBs) or against the core antigen (anti-HBc). Over all age-groups, the prevalences of HBsAg, anti HBs and anti-HBc were 0.67%, 71.4% and 5.5%, respectively. Although the prevalence of anti-HBs decreased with age, it remained at 56%-65% among those aged 6-10 years. Between 2% and 17% of the subjects aged 1-9 years had high titres of anti-HBs. Based on these results, an additional booster, still a controversial issue, does not appear to be required in order to prevent infection with hepatitis B virus and thus permit the eventual eradication of chronic carriage and its fatal sequelae in Thailand. PMID- 11064764 TI - Lutzomyia whitmani (Diptera: Psychodidae) as vector of Leishmania (V.) braziliensis in Parana state, southern Brazil. AB - The phlebotomine sandflies in the northern areas of the state of Parana, Brazil, particularly those in the '16a' health region, were investigated over a 3-year period. Using CDC light traps (with and without hamster bait) and Shannon traps (with lights and horse or human bait), 16 species were collected from seven municipal districts which were known foci for cutaneous leishmaniasis: Arapongas; Apucarana; Cambira; Marumbi; Faxinal; Florestopolis; and Sabaudia. Although the frequency at which each species was collected varied with the collection site, Lutzomyia whitmani predominated (62.0% of all the sandflies collected), followed by Lu. fischeri (13.3%), Lu. pessoai (10.8%), Lu. migonei (8.2%) and Lu. intermedia (2.8%). Lutzomyia monticola, Lu. shanonni, Lu. firmatoi, Lu. lanei, Lu. alphabetica, Lu. misionensis, Lu. correalimai, Lu. cortellezzii, Lu. longipenis, Brumptomyia brumpti and B. nitzulescui together represented the remaining 3.0% of the collected sandflies. Three of the 1961 female sandflies collected and dissected in the municipal district of Cambira, where a recent case of cutaneous leishmaniasis had been registered, were found to have flagellates in their guts. All three were Lu. whitmani. The parasites from each of these infections were successfully isolated in NNN and 'Tobie and Evans' media and/or by inoculation into a hind foot of a golden hamster. The results of isoenzyme electrophoresis indicated that all three isolates were of Leishmania (Viannia) braziliensis. PMID- 11064765 TI - Effects of Triatoma virus (TrV) on the fecundity and moulting of Triatoma infestans (Hemiptera: Reduviidae). AB - Triatoma virus (TrV) is a Picorna-like virus affecting Triatoma infestans (Klug, 1834), the most important transmitter of Trypanosoma cruzi in South America. The subjects of the present, laboratory study were the longevity and oviposition of female Tri. infestans, from stocks with and without viral infection, and the survivorship and developmental time of their progeny through to second-instar nymphs. On average, adult females from an infected stock lived only a third as long as those from the uninfected, and the mean monthly egg outputs of the 'infected' females was only 20% of that of the uninfected females. Even though the virus can be transmitted transovarially and most, if not all, of the progeny of the infected females were themselves infected, there was no evidence for TrV affecting egg hatchability. A much smaller proportion of the progeny of the females from infected stock than of that of the uninfected females successfully moulted to second-instar nymphs (44% v. 80%); the virus appears to inhibit the moulting process. The progeny of the females from the infected stock developed relatively slowly, spending a mean of 6.1 days as first instars (compared with 3.9 days for the progeny of uninfected females). Together, these data indicate that TrV may be a useful agent for the biological control of Tri. infestans. PMID- 11064766 TI - Exploration of the distribution of Trypanosoma brucei ssp. in West Africa, by multilocus enzyme electrophoresis. PMID- 11064767 TI - Internal-transcribed-spacer (ITS) sequences used to explore phylogenetic relationships within Leishmania. PMID- 11064768 TI - Hypernatraemia in early infancy. AB - Hypernatraemia, defined as serum sodium > 150 mmol/l, is still seen frequently in tropical environments. We describe two infants, one with poor fluid intake and excessive wrapping and the other with a high solute intake, both of whom presented in a state of severe hypernatraemia. The pathophysiology of this condition is outlined, as are the major causes, such as sodium overload, inadequate water intake, increased water loss of non-renal origin, increased water loss of renal origin and essential hypernatraemia. The literature is reviewed and the current basis for management is appraised and discussed. PMID- 11064769 TI - Malnutrition and hypernatraemia in breastfed babies. AB - Despite the well-known advantages of breast-feeding to both mother and infant, malnutrition of breastfed infants does occur. We report two term neonates who presented in the 3rd week of life with severe wasting, hypernatraemic dehydration and pre-renal failure while being exclusively breastfed. Breast-milk sodium levels were markedly elevated on admission. Both infants recovered following adequate hydration and showed excellent catch-up growth during follow-up while exclusive breast-feeding was maintained. The critical malnutrition in both cases was detected by the family physician during routine postnatal visits. Both mothers were well motivated toward breast-feeding and were unaware of the severity of the baby's illness. PMID- 11064770 TI - Monotherapy in an era of combination therapy: is there a benefit? Experience in HIV-1-infected symptomatic South African children. AB - We investigated the benefit of treating HIV-1-infected children with monotherapy where resources are limited. A retrospective chart review was undertaken in 12 symptomatic HIV-1-infected children treated with zidovudine or didanosine for at least 2 months. The main outcome measure was the effect on hospitalization. Anti retroviral therapy was commenced in nine children because of prolonged or frequent hospitalization. Of three whose primary indication was bleeding secondary to thrombocytopenia, two had been hospitalized owing to severe intercurrent illness. One child had failure to thrive and another encephalopathy. Monotherapy was considered beneficial in all cases. Median duration of follow-up was 6.5 (2-31) months. The hospitalization index (days in hospital before and after start of monotherapy, divided by the total number of days before and after start of monotherapy) decreased from a median of 0.115 prior to therapy to 0.037 on therapy (p = 0.045, Wilcoxon matched pairs test). This study presents observational data supporting the investigation of monotherapy in resource-poor countries. It was associated with a significant reduction in hospitalization and appeared to result in clinical improvement. Prolonged or frequent hospitalization might represent a novel indication for use because in our setting the cost of hospitalization could potentially provide a 12-month-old infant with monotherapy for 2 months. PMID- 11064771 TI - Bacterial infection in children with HIV: a prospective study from Cape Town, South Africa. AB - Invasive bacterial infection in children infected with the human immunodeficiency virus (HIV) is common. South African data on this problem are limited. Over 1 year we prospectively studied 108 HIV-infected children hospitalized for 136 presumed infective episodes. Blood culture was positive in 24.8% of episodes. Streptococcus pneumoniae predominated (14/30 positive blood cultures); one-third of isolates showed resistance to penicillin. Acute lower respiratory tract infection accounted for 44% of clinical diagnoses, a bacterial cause being established for 23.8% of these. Age and stage of HIV infection did not influence the likelihood of a positive culture. A high proportion of presumed infective episodes requiring hospitalization of young HIV-infected children have a bacterial cause. Blood culture appears to be a useful method of obtaining the microbiological information required to focus antibiotic therapy. PMID- 11064772 TI - A comparison of the efficacy of cefuroxime axetil and intramuscular benzathine penicillin for treating streptococcal tonsillopharyngitis. AB - Throat cultures were performed on 297 children suspected of tonsillopharyngitis on clinical findings. Group A beta-haemolytic streptococci (GABHS) were isolated from 86 patients (41 males/45 females) aged 6-15 (mean (SD) 7.8 (0.04)) years. They were randomly allocated to receive oral cefuroxime axetil for 10 days (group 1) or one dose of benzathine penicillin by intramuscular injection (group 2) and responses were evaluated 2 weeks later. Clinical cure was observed in 95% of group 1 and 96% of group 2 and bacteriological cure in 86 and 84% of groups 1 and 2, respectively. Our results show that intramuscular benzathine penicillin remains an effective treatment for GABHS and that oral cefuroxime axetil is also effective. PMID- 11064773 TI - Cystic fibrosis in a large kindred family in Qatar. AB - We describe 45 patients with cystic fibrosis (CF), diagnosed between June 1987 and May 1999, seen at the Hamad Medical Corporation, Qatar in the Arabian Gulf. Twenty-six of 32 families in the study were related and belonged to the same Bedouin tribe. The parents of 98% of these cases were consanguineous. Metabolic alkalosis and/or hypo-electrolytaemia were found in a large proportion of infants with CF. Cystic fibrosis in Qatari children is phenotypically variable with mild to moderate respiratory symptoms, and none of them died during this study. Among the non-Arabic-Asian patients, pulmonary symptoms were more severe, Pseudomonas colonization was earlier, pancreatic insufficiency occurred in infancy and four died in early life. PMID- 11064774 TI - Diphtheritic myocarditis: clinical and laboratory parameters of prognosis and fatal outcome. AB - Of the 97 throat smear-positive cases of diphtheria treated, 16 developed evidence of myocarditis (16.5%). Age, duration of illness prior to admission, anatomical extent of membrane formation, total leukocyte count and serum glutamic oxalo-acetic transaminase (SGOT) levels were analysed in fatal and non-fatal cases and the differences between the two groups found to be statistically significant with regard to anatomical extent of membrane formation, total leukocyte count and SGOT levels. The extension of membrane formation to two or more sites was a highly sensitive predictor of mortality. A total leukocyte count > 25,000 cells/mm3 had a high specificity and positive predictive value while SGOT levels of > 80 IU/l had high sensitivity and a negative predictive value. PMID- 11064775 TI - Case-control study of diet and sun exposure in adolescents with symptomatic rickets. AB - Dietary intake and sun exposure were compared in a case-control study of 14 adolescent girls diagnosed to have symptomatic nutritional rickets with hypocalcaemia, hypophosphoraemia, elevated serum alkaline phosphatase and serum parathormone and reduced 25-hydroxy vitamin D levels and 20 controls without clinical rickets (ten girls) of the same age group and socio-economic background. In the control group, calorie intake was reduced in seven boys and eight girls, dietary calcium in seven boys and seven girls and vitamin D in six boys and eight girls. All 20 adolescents of both sexes were exposed to the sun for more than 60 min a day. In the group with rickets, calorie intake was reduced in 11 girls, calcium in 14 girls and vitamin D in nine girls). Sun exposure was significantly less in girls with rickets than in controls (p < 0.001). Adolescents in our population, especially females, are at high risk of developing nutritional rickets. Prevention by longer exposure to the sun is simple and cheap but when not practical for social or cultural reasons routine vitamin D supplementation might be indicated. PMID- 11064776 TI - A retrospective review of paediatric lymph node tuberculosis in Port Moresby, Papua New Guinea. AB - The study aimed to examine the perception that the relapse rate following standard short-course chemotherapy in children with lymph node tuberculosis is greater than in similarly treated children with pulmonary tuberculosis (TB). The treatment records, clinical data and results of investigations of 427 children treated for lymph node TB between 1989 and 1996 were analysed. The results and role of investigations are discussed. The relapse rate was compared with that of 892 children treated for pulmonary TB during the same period. The documented relapse rate for lymph node TB was 2.8% compared with 0.6% in pulmonary TB. This highly significant difference led to a prospective study of outcome of lymph node TB treatment in Port Moresby. PMID- 11064777 TI - Febrile convulsions: acute seizure characteristics and anti-convulsant therapy. AB - A descriptive study using data from the medical records of 448 children with febrile convulsion was carried out to determine the seizure characteristics and use of anti-convulsant therapy for febrile convulsions in a Malaysian hospital. There was a higher incidence of multiple seizures and a lower incidence of focal seizures in the local population than in studies done among Western populations. The majority of initial seizures occurred within 24 h of fever onset. Transient neurological abnormalities following an acute seizure were common. A quarter of children referred by general practitioners had been given anti-convulsants prior to referral but up to 20% of general practitioners had used ineffective routes for administering diazepam. However, diazepam used in the hospital was found to be effective in controlling acute febrile seizures. PMID- 11064778 TI - An epidemic of congenital rubella in Barbados. AB - Rubella and congenital rubella syndrome (CRS) are preventable, but epidemics of rubella and CRS are not infrequent in the Caribbean and other developing countries. As a result of a surveillance system initiated after an epidemic of rubella in the Barbadian population in 1996, cases of CRS were identified and investigated. A total of seven cases of CRS were proven to be rubella IgM positive. The infants were found to have a mean birthweight of 2587 g and a mean gestational age of 38 weeks. The clinical course, complications and outcome of those infants were documented and the cost of acute hospital care for each patient was also recorded. Cataracts in four infants, congenital heart disease in three and central nervous system abnormalities in five were the major clinical abnormalities. In four infants, two or more clinical systems were affected. The combined total hospital stay was 105 days (mean 15, range 0-44). A national effort to immunize all those at risk and a strict surveillance programme are essential to prevent future epidemics. This would lead to a significant reduction in the number of cases of rubella and CRS and could effect substantial savings in the national health budget. PMID- 11064779 TI - Absence of bone marrow amastigotes in a child with kala-azar and acute lymphoblastic leukaemia. AB - Visceral leishmaniasis is suspected on the basis of clinical findings and a pancytopenic blood picture and is usually confirmed by the detection of amastigotes (Leishman-Donovan bodies) in a bone marrow aspirate. We describe a child on maintenance treatment for acute lymphoblastic leukaemia who developed visceral leishmaniasis and in whom amastigotes could not be detected in repeated bone marrow aspirates. Immunofluorescence antibody testing was positive. Immunocompromised patients in endemic areas who develop features of visceral leishmaniasis should have serological tests performed in addition to bone marrow aspiration in order to maximize the chances of making a diagnosis. PMID- 11064780 TI - Spina bifida cystica: selective management in Zaria, Nigeria. AB - Management of spina bifida cystica in Zaria, Nigeria is selective. Over a period of 11 years, 77 children with this defect, 54 meningocoeles and 23 myelomeningocoeles, 66 (86%) situated in the lumbosacral region, were treated operatively. Forty-two (55%) had surgery in the neonatal period and 91% within 6 months of birth. Postoperative complications occurred in 19 of 68 patients (28%), including mild hydrocephalus, which resolved spontaneously (six, 9%), wound infection (six, 9%), leakage of cerebrospinal fluid (four, 6%) and meningitis (three, 4%). Mortality was 3% from both meningitis and cardiac arrest. Of 32 patients followed up for 3-5 years, 20 with meningocoeles were normal. Of 12 with myelomeningocoele, four had varying degrees of lower limb weakness, three double incontinence, two faecal incontinence, two had progressive hydrocephalus plus paralysis and double incontinence, and one had urinary incontinence. Therefore, 38% were functionally disabled and could not be adequately rehabilitated owing to poor facilities. While management of spina bifida cystica is more aggressive now in most developed countries, ours remains selective due to difficulty with multidisciplinary care and rehabilitation. Even with our selective management, the care of patients with functional handicap remains a challenge. PMID- 11064781 TI - Cardiovascular function in Omani children with sickle cell anaemia. AB - Systolic murmurs were detected in 22 (61%) of the 36 children with sickle cell anaemia (SCA) who completed the study. Cardiomegaly was detected in 14 (39%). Mean values of left and right ventricular dimensions were higher in SCA than in controls (p < 0.05). Left atrial chambers and aortic root dimensions followed the same pattern. The dilated cardiac chambers in SCA were not associated with any abnormality in systolic or diastolic left ventricular function nor with significant pulmonary hypertension. PMID- 11064782 TI - Heart failure now on the palliative care agenda. PMID- 11064783 TI - Survival prediction in terminal cancer patients: a systematic review of the medical literature. AB - The clinical significance of studies on survival predictors in terminal cancer patients is hindered by both methodological limitations and the difficulty of finding common predictors for all final events in cancer related deaths. To evaluate the published medical literature concerned with the survival of patients with terminal cancer and identify potential prognostic factors, major electronic databases including MEDLINE (1966-), CANCERLIT (1983-) and EMBASE (1988-) were searched up to September 1999. Studies were included in our review if published in English, were cohort studies, addressed the identification of clinical prognostic factors for survival and looked at samples with median survival of < or = 3 months. Data extracted from selected papers included: sample size, median survival, type of study, sampling frame, cohort type, type of statistical analysis (univariate or multivariate), choice of models and underlying assumptions, predictors examined and their reported level of statistical significance. A total of 24 studies were found and reviewed. On the basis of these studies, performance status and the presence of cognitive failure, weight loss, dysphagia, anorexia and dyspnoea appear to be independent survival predictors in this population. Clinical estimation of survival by the treating physician appeared independently associated with survival but the magnitude of the association generally appeared small. Clinical predictions should be considered as one of many criteria, rather than as a unique criterion by which to choose therapeutic interventions or health care programmes for terminally ill cancer patients. The use of convenient samples as opposed to more representative inception cohorts, the inclusion of different variables in the statistical analyses and inappropriate statistical methods appear to be major limitations of the reviewed literature. Methodological improvements in the design and conduction of future studies may reduce the prognostic uncertainty in this population. PMID- 11064784 TI - A randomized controlled trial of a hospital at home service for the terminally ill. AB - This study evaluated the impact of a Cambridge hospital at home service (CHAH) on patients' quality of care, likelihood of remaining at home in their final 2 weeks of life and general practitioner (GP) visits. The design was a randomized controlled trial, comparing CHAH with standard care. The patient's district nurse, GP and informal carer were surveyed within 6 weeks of patient's death, and 225 district nurses, 194 GPs and 144 informal carers of 229 patients responded. There was no clear evidence that CHAH increased likelihood of remaining at home during the final 2 weeks of life. However, the service was associated with fewer GP out of hours visits. All respondent groups rated CHAH favourably compared to standard care but emphasized different aspects. District nurses rated CHAH as better than standard care in terms of adequacy of night care and support for the carer, GPs in terms of anxiety and depression, and informal carers in terms of control of pain and nausea. Thus whilst CHAH was not found to increase the likelihood of remaining at home, at appeared to be associated with better quality home care. PMID- 11064785 TI - The reality of palliative care in Spain. AB - With the sponsorship of the Spanish Society of Palliative Care (SECPAL), two nation-wide studies from the University of Valladolid were carried out in 1996 and 1998 into the state of palliative care teams in Spain. This report is based on those studies and is the first overall analysis of the current situation of palliative care in Spain. A total of 143 programmes were identified and data were collected from 128 teams (89%): 53 were programmes with hospitalised patients, 75 for home-based patients and 15 were mixed programmes. From the activity data, provided by 75% of the teams, it may be estimated that each year 22,638 terminally ill patients receive palliative care attention. Of these, 18,021 (79.6%) are cancer sufferers. Of the patients who die from cancer each year in our country, 21.2% receive palliative care during the final weeks of life. More than 10 years have passed since the first teams in Spain started their work. The time has now come to recognize the reality of palliative care in our society and to adopt a national policy for terminally ill patients, in line with World Health Organization recommendations formulated in 1989. PMID- 11064786 TI - Minimum dataset activity for hospice and hospital palliative care services in the UK 1997/98. AB - This study reports on the third in an annual series of surveys covering England, Wales, Scotland and Northern Ireland on the activity of palliative care services. This report concentrates on inpatient (hospice and hospital) services. All 640 known UK palliative services were sent a standardized questionnaire asking about the characteristics and numbers of patients cared for. Results were analysed for those services primarily for adults. From the 189 inpatient units (2955 beds) there was an 84% response rate in total, but the response to many of the questions was lower than this. Sixty per cent of services recorded 24,362 new patients, and about 50% provided details showing that 96.7% patients had cancer, and one-third were aged under 65 years. This is higher than the national distribution of cancer deaths where 24% are under 65 years. Conversely, only 7% were in the over 84 age group, which has 14% of cancer deaths. Most patients (73%) were admitted from their own home. Half of the admissions ended in death, and the majority of discharges were to the patient's own home. The mean length of stay was 13.1 days, with larger units tending to have a longer length of stay. Forty per cent of admissions were for one week or less (2.3% of patients died on the day of admission) and 15% were for more than three weeks. Bed occupancy varied between 99.7 and 48.9%. Responses were received from 74% of the 326 hospital support services, although again many questions were answered by less than half of those eligible to do so. Details of 37,194 new patients were reported (5.9% did not have cancer, although there was a wide range between services). Patients typically had four contacts with the service, although almost a quarter were single contacts. Three-quarters of the contacts were by a clinical nurse specialist. National estimates suggest that of the 155,000 patients dying of cancer in the UK each year 27,600 (18%) die in a hospice. There are 39,000 new hospice admissions each year and about 100,000 patients have contact with a hospital support service. Overall, the national provision of palliative care is increasing but there are groups who still appear to be missing out on palliative care, especially older people. Increasingly, patients appear to be admitted to a hospice earlier in care and are discharged home. PMID- 11064787 TI - What do palliative medicine consultants do? AB - A workload study of consultants in palliative medicine was carried out, in which a task inventory questionnaire was sent to the consultant members of the Association for Palliative Medicine of Great Britain and Ireland. The median number of hours per week worked by a palliative medicine consultant was 49.5 (mean 51.3), excluding time on-call and breaks. The median consultant first on call roat is 1:3. Palliative medicine consultants work a median of 22.5 direct clinical and 4.5 indirect clinical hours per week. This is very similar to the hours worked by consultants in other specialties who work an average 51.3 h per week, but with a median on-call rota of 1:6. Further studies are planned to monitor the patterns of consultant activity and workload in this specialty. PMID- 11064788 TI - Perceptions of health care need in lung cancer. Can prospective surveys provide nationally representative data? AB - This paper considers the potential of a prospective questionnaire survey to provide nationally representative data of perceptions of health care need. The paper focuses on methodological problems encountered during the study, reserving the discussion of key findings to further publications. Data were gathered from patients with lung cancer, lay carers and professionals from 24 randomly selected hospitals throughout the UK. A number of factors had an impact on the survey's potential to provide nationally representative data. These included an unanticipatedly high death rate of patients registered in hospitals as being in receipt of treatment or follow-up care. Of a potential sample of 785 patients identified as being alive by the participating hospitals prior to data collection, 319 (41%) were found to have died after checking with general practitioner surgeries. As a consequence of consultants declining to participate, or where they did not reply to any correspondence, eight hospitals were withdrawn from the study on the grounds of inability to gain access to a representative sample of patients. In conclusion, a retrospective design, supplemented by the views of patients would have ensured greater success in terms of patient response rate (45%) and representativeness of data. Nevertheless, the survey represents a first attempt at undertaking a national assessment of patients with lung cancer and has provided rich data drawn from patients' experiences of living with a diagnosis of lung cancer and its treatment. PMID- 11064789 TI - Participation in a creative arts project can foster hope in a hospice day centre. AB - This study explored the experiences of terminally ill patients taking part in an exhibition of their creative arts work. It took place in St Christopher's Hospice day centre, London, UK, which aims to facilitate an environment in which a range of social and creative opportunities is offered following the theoretical background of Maslow's and Rogers' theories of personal growth and creativity. A phenomenological study explored the views of 10 patients and eleven facilitators using in-depth, semi-structured, audiotaped interviews. A content analysis identified the main themes as enjoyment, enthusiasm, excitement, pride, achievement, satisfaction, sense of purpose, mutual support and permanence. These themes were interpreted as positive expressions of self-esteem, autonomy, social integration and hope. It is suggested that it was possible to identify hope as the essence of the phenomenon, and that this is important in palliative care where traditionally continuation of active medical intervention has been equated with provision of hope. PMID- 11064790 TI - The involvement of specialist palliative care in the care of people with motor neurone disease. PMID- 11064791 TI - The use of thalidomide in the management of severe sweating in patients with advanced malignancy: trial report. PMID- 11064792 TI - Complementary therapy use: a survey of community- and hospital-based patients with advanced cancer. PMID- 11064793 TI - Why are trials in palliative care so difficult? PMID- 11064794 TI - Midazolam-induced dyskinesia. PMID- 11064795 TI - Survival of communal rituals in rural communities. PMID- 11064796 TI - Cultural attitudes towards death and dying: a South African perspective. PMID- 11064797 TI - Support and supervision in palliative care research. PMID- 11064798 TI - Society, science, and values. PMID- 11064799 TI - The minimum temporal thresholds for motion detection of grating patterns. AB - The minimum temporal thresholds for absolute motion detection were measured for sinusoidal grating patterns in foveal vision. Test patterns of relatively low temporal frequencies and low velocities were examined. The thresholds clearly decreased with test velocities rather than with test temporal frequencies. Modified velocity-time reciprocity was observed (i.e. the relationship between test velocity and temporal thresholds was described by a simple equation including two constants which indicate temporal and spatial limits). The temporal constant was about 35 ms and the spatial constant was about 1 min of arc. These constants are thought to provide the basic constraints on motion detection. PMID- 11064800 TI - Visual motion integration for perception and pursuit. AB - To examine the relationship between visual motion processing for perception and pursuit, we measured the pursuit eye-movement and perceptual responses to the same complex-motion stimuli. We show that humans can both perceive and pursue the motion of line-figure objects, even when partial occlusion makes the resulting image motion vastly different from the underlying object motion. Our results show that both perception and pursuit can perform largely accurate motion integration, i.e. the selective combination of local motion signals across the visual field to derive global object motion. Furthermore, because we manipulated perceived motion while keeping image motion identical, the observed parallel changes in perception and pursuit show that the motion signals driving steady-state pursuit and perception are linked. These findings disprove current pursuit models whose control strategy is to minimize retinal image motion, and suggest a new framework for the interplay between visual cortex and cerebellum in visuomotor control. PMID- 11064801 TI - Attenuation of alignment effect with exocentric encoding of location. AB - An object's location is best retrieved from the orientation in which it was learned. Otherwise, retrieval necessitates a mental effort to restore the original perspective. In this case there is a cost to speed and accuracy of location responses known as the alignment effect. We hypothesised that one can attenuate this alignment effect by systematically referring objects in an exocentric frame of reference during learning. Sixteen male students were asked to learn the location of five objects disposed in a totally new environment either by locating the objects in an egocentric or in an exocentric spatial frame of reference. After the learning phase, the participants were asked to imagine orienting themselves to an object in the scene and to point to another object. The analysis of pointing accuracy, orientation, and pointing times showed that the performances of participants engaged in the exocentric condition remained insensitive to the augmentation of the angle between their actual position on the path and the imagined orientation. On the other hand, the participants engaged in egocentric learning were disoriented when the difference between their actual orientation and the imagined orientation was great. We conclude that when an object's location is intentionally referred to in an exocentric reference frame, alignment effect can be significantly reduced. PMID- 11064802 TI - Optic flow helps humans learn to navigate through synthetic environments. AB - Self-movement through an environment generates optic flow, a potential source of heading information. But it is not certain that optic flow is sufficient to support navigation, particularly navigation along complex, multi-legged paths. To address this question, we studied human participants who navigated synthetic environments with and without salient optic flow. Participants used a keyboard to control realistic simulation of self-movement through computer-rendered, synthetic environments. Because these environments comprised series of identically textured virtual corridors and intersections, participants had to build up some mental representation of the environment in order to perform. The impact of optic flow on learning was examined in two experiments. In experiment 1, participants learned to navigate multiple T-junction mazes with and without accompanying optic flow. Optic flow promoted faster learning, mainly by preventing disorientation and backtracking in the maze. In experiment 2, participants found their way around a virtual city-block environment, experiencing two different kinds of optic flow as they went. By varying the rate at which the display was updated, we created optic flow that was either fluid or choppy. Here, fluid optic flow (as compared with choppy optic flow) enabled participants to locate a remembered target position more accurately. When other cues are unavailable, optic flow can be a significant aid in wayfinding. Among other things, optic flow can facilitate path integration, which involves updating a mental representation of place by combining the trajectories of previously travelled paths [corrected]. PMID- 11064803 TI - Does time-shrinking take place in visual temporal patterns? AB - The duration of a short empty time interval (typically shorter than 300 ms) is often underestimated when it is immediately preceded by a shorter time interval. This illusory underestimation--time-shrinking--had been studied only with auditory temporal patterns. In the present study, we examined whether similar underestimation would take place with visual temporal patterns. It turned out that underestimation of the same kind takes place also in the visual modality. However, a considerable difference between the auditory and the visual modalities appeared. In the auditory modality, it had been shown that the amount of underestimation decreased for preceding time intervals longer than 200 ms. In the present study, the underestimation increased when the preceding time interval varied from 160 to 400 ms. Furthermore, the differences between the two neighbouring intervals which could cause this underestimation had always been in a fixed range in the auditory modality. In the visual modality, the range was broader when the intervals were longer. These results were interpreted in terms of an assimilation process in light of the processing-time hypothesis proposed by Nakajima (1987 Perception 16 485-520) in order to explain an aspect of empty duration perception. PMID- 11064804 TI - The temporal course of suppression during binocular rivalry. AB - Orthogonally oriented sinusoidal luminance gratings were dichoptically presented to the observers' left and right eyes. During the subsequent binocular rivalry, a small target was briefly presented (4AFC) to probe the strength of interocular suppression at various temporal latencies. Both stationary and moving rivalrous patterns were investigated. The purpose of experiment 1 was to compare the temporal characteristics of stationary and motion rivalry (0 and 1.2 deg s-1), while that of experiment 2 was to examine rivalry suppression for higher speeds (2 and 4 deg s-1). In all cases, it was found that the strength of suppression remained essentially constant throughout a single phase of binocular rivalry. The results of the investigation also revealed that moving rivalrous patterns lead to greater magnitudes of interocular suppression than static patterns. Despite these differences in the strength of suppression, the results of both experiments show that the temporal characteristics of motion and static rivalry are essentially identical. PMID- 11064805 TI - Form and movement in stereokinetic cycloids: motion lost and found. AB - In exploring stereokinesis, we devised flat cycloidal display figures which, when rotated on a disc in the frontal plane, are perceived as illusory three dimensional forms with movement in depth; the dominant percepts were of twisted loops with an internal writhing motion. These dominant forms could be convincingly represented by stereo pairs derived from the flat display; related forms, not seen in the illusion, could also be constructed, seeming to show a selectivity for preferred stereokinetic forms by the perceptual system. Models were made of the stereo forms; when rotated, they showed similar illusions and selectivity. We suggest that the illusions arise because some components of the real motion do not appear in the sensory field. The perceptual system accommodates for this by constructing percepts which are not necessarily veridical but do reconcile form and motion into a coherent unity. The results are discussed in relation to concepts of invariance and rigidity, and with regard to the creative response to sensory data by the perceptual system. PMID- 11064806 TI - Luminance determinants of perceived surface stratification in two-dimensional achromatic transparent patterns. AB - Two overlapping transparent surfaces forming a two-dimensional pattern stand out in front of each other alternately. Let y denote the luminance of the region where these surfaces overlap and b the luminance of the background. In achromatic patterns, the probability that the lighter transparent surface appears to be in front of the other surface is known to increase with y and with b. The present results show that grouping by achromatic colour similarity cannot explain the effect of b. An alternative conjecture is that the luminance factors that control perceived surface segregation can explain the effects of y and b. Such an explanation predicts a new effect: the probability that one transparent surface appears to be in front increases with the absolute difference in luminance between the surface and the background. The present results confirm this prediction. PMID- 11064807 TI - Cleomedes (c. 1st century AD) on the celestial illusion, atmospheric enlargement, and size-distance invariance. AB - Cleomedes (Kleomedes) is a little-known Greek author (c. 1st century AD) who produced what is probably the earliest extant statement of size-distance invariance. He supported the Stoic philosophy and was concerned to discredit the Epicurean position that we perceive objects as having their true size. He explained the celestial illusion (the apparent enlargement of the sun near the horizon) in two ways: partly as a refractive effect of the atmosphere similar to angular enlargement when looking into water; and partly as a linear enlargement due to increased apparent distance in a misty atmosphere. He is the earliest extant author to offer apparent distance as a clear explanation of the celestial illusion. He attributed these views to Posidonius (c. 135-50 BC). His explanations remained at the geometrical level, and he did not speculate on sensory mechanisms. PMID- 11064808 TI - Last but not least. Perception. PMID- 11064809 TI - The sawtooth illusion. PMID- 11064810 TI - Diagnostic utility of BCA-225 in detecting adenocarcinoma in serous effusions. AB - OBJECTIVE: To determine the diagnostic value of the BCA-225 antibody in discriminating adenocarcinoma from benign mesothelium in body cavity effusions. STUDY DESIGN: One hundred four cases of unequivocally benign (34 cases) and malignant (70 cases) serous effusions with cell block material were immunostained for BCA-225 using the ABC method without antigen retrieval. The percentage of positively staining cells in each case was estimated in a blind fashion. RESULTS: BCA-225 stained at least 10% of morphologically malignant cells in 28 of 32 (88%) breast carcinomas and 58 of 67 (87%) adenocarcinomas overall. Neuroendocrine carcinomas (two cases) and one mesothelioma were positive in < or = 5% of their respective tumor cells. Of 34 benign cases, 6 (18%) exhibited positive staining, albeit in rare, morphologically benign cells. CONCLUSION: BCA-225 is able to discriminate adenocarcinoma from reactive mesothelium in cell block preparations and may prove useful as part of an antibody panel. PMID- 11064811 TI - Cell cycle-dependent AgNOR analysis in invasive breast cancer. AB - OBJECTIVE: To investigate to what extent analysis of silver-stained nucleolar organizer regions (AgNORs) is cell cycle dependent in breast cancer and to assess the prognostic value of an AgNOR analysis that takes into consideration the cell cycle status of tumor cells. STUDY DESIGN: In 97 cases of invasive breast carcinoma, morphometric AgNOR analysis was performed in tumor cells with immunohistochemical MIB-1 reactivity (NORcyc analysis) and in MIB-1-negative tumor cells (NORnon analysis). Additionally, conventional (NORconv) analysis without preceding MIB-1 staining was done. Findings were compared with the Nottingham prognostic index (NPI). RESULTS: In comparison to noncycling tumor cells, cycling ones exhibited significantly higher AgNOR numbers (mean values, 3.84 +/- 1.09 vs. 2.40 +/- 0.78 per nucleus), higher total AgNOR areas (5.95 +/- 3.17 vs. 5.62 +/- 3.05 micron 2, NS) and significantly lower mean AgNOR areas (2.08 +/- 1.14 vs. 2.93 +/- 1.69 micron 2). When related to NPI, correlation coefficients of NORnon analysis were higher than those of NORcyc analysis but lower than those of NORconv analysis. Among the different AgNOR parameters, total AgNOR area correlated best with NPI. CONCLUSION: Cell cycle status has a high impact on AgNOR analysis. However, the best prognostic information in breast cancer is derived from an AgNOR analysis that considers both cycling and noncycling tumor cells. PMID- 11064812 TI - Integrated nuclear fluorescence and expression of hormone-binding sites in malignant pleural effusions. AB - OBJECTIVE: To investigate potential disease- and prognosis-associated nuclear and cellular features from cell properties in a prospective study on malignant pleural effusions. STUDY DESIGN: Integrated nuclear fluorescence and the expression of binding capacities of carrier-immobilized estradiol, progesterone and testosterone; and of labeled sarcolectin; and the presence of calcyclin were measured in 50 cases with proven malignant pleural effusions (10 mesotheliomas, 40 metastasizing tumors). A double fluorescence technique using the fluorochrome DAPI and a Texas Red-based avidin-biotin detection system were applied. Detailed clinical data, including the follow-up for up to 40 months, were included. RESULTS: Pleural effusions in all patients with mesotheliomas occurred prior to (9/10) or at the time of histologic confirmation. Mesotheliomas had the highest tumor cell fraction (12.4%) in S phase and breast carcinomas the lowest (10.7%). More than 80% of malignant cells expressed binding capacities for the applied probes. A statistically significant correlation was noted between the S-phase related tumor cell fraction and the expression of progesterone receptors. Survival was associated with tumor origin, treatment by pleurodesis, and certain cytometric and histochemical features. CONCLUSION: The immunofluorescence double staining technique can be applied successfully in malignant effusions to combine DNA measurements with those of immunohistochemical and ligand histochemical reactivity. PMID- 11064813 TI - Validation of nuclear texture, density, morphometry and tissue syntactic structure analysis as prognosticators of cervical carcinoma. AB - OBJECTIVE: To evaluate the performance of karyometry and histometry in the prediction of survival, recurrence and response of early-stage invasive cervical carcinoma. STUDY DESIGN: Nuclear morphometry, chromatin texture and tissue architecture (characterized by syntactic structure analysis) were measured using a semiautomated image analysis system on 46 cases of Feulgen-stained tissue sections. The performance of the features was compared to that of clinical features, reported to be the best prognosticators until now, such as age, lympho vascular permeation, histologic type, stage and grade. A K nearest neighbor classifier was used for classification. RESULTS: In the prediction of three-year survival, recurrence and response, syntactic structure analysis proved to be the best performer. Classification rates were, respectively, 100%, 94.4% and 94.5%. In all classifications, karyometric and histometric features outperformed clinical features. In general, the best performing features described differences in second-order population statistics (standard deviations). CONCLUSION: The results show that a quantitative analysis based on nuclear morphology, chromatin texture and histology can be considered an excellent aid in the prognosis of invasive cervical carcinoma. The measurements are not hampered by the need to undertake complete resections and are suited to daily practice when implemented in a semiautomated image analysis system. PMID- 11064814 TI - Diagnosis of human oligodendrogliomas with the help of the NeuroShell Easy Classifier neural network. AB - OBJECTIVE: To examine whether a suitable solution can be found concerning the ability to reproduce the histologic classification of human oligodendrogliomas with the assistance of the NeuroShell Easy Classifier neural network. STUDY DESIGN: Histologic sections of 449 human oligodendrogliomas were selected. The diagnostic task was given by differentiation of three oligodendroglioma types: 121 low grade oligodendrogliomas, World Health Organization grade 2; 180 low grade oligoastrocytomas; and 148 anaplastic oligodendrogliomas, grade 3. Age, sex and 50 histologic characteristics were examined in each case, describing the presence of a specific histologic feature on a scale of four (zero, absence of the feature; three, abundant presence). From each group, two-thirds of randomly selected tumors were available for the training set and one-third for the testing set. RESULTS: In the three-class problem, 98.88% of the tumors were correctly classified (testing set). Ninety-nine percent of new testing tumors were correctly classified with Easy Classifier as low grade and anaplastic oligodendrogliomas. In the case of low grade oligodendrogliomas versus low grade oligoastrocytomas, 99% of new tumors were correctly classified. CONCLUSION: The main conclusion from this study is that Easy Classifier was able to differentiate, with high accuracy, sensitivity and specificity, among the three types of oligodendrogliomas. PMID- 11064815 TI - Proliferation and apoptosis in solid tumors. Analysis by laser scanning cytometry. AB - OBJECTIVE: To examine the relationship between apoptosis and proliferation in a series of human solid malignant tumors, making use of objective, reproducible techniques newly developed for laser scanning cytometry (LSC). STUDY DESIGN: Apoptosis was detected by in situ end labeling of DNA strand breaks with FITC conjugated nucleotide. Proliferation was detected by Ki-67 antibody. Two parameters were detected independently and simultaneously with DNA measurement on aliquots of cell suspensions obtained by mechanical dissociation of fresh tumors and placed on microscope slides. RESULTS: The number of cells undergoing apoptosis varied from 0.5% to 28.1% (average, 5.4 +/- 6.0). Aneuploid tumors showed a higher percentage of apoptotic cells (7.9 +/- 7.2) as compared to diploid tumors (3.4 +/- 4.0). Tumors with the greatest number of apoptotic cells on LSC also had the largest number of apoptotic cells on light microscopic examination. The number of cells labeled by Ki-67 ranged from 1.7% to 56.7% (average, 20.0 +/- 15.5). Aneuploid tumors were characterized by a higher Ki-67 index (average, 28.3 +/- 14.3%) than the diploid tumors (13.2 +/- 13.3%). CONCLUSION: Overall, there was a very weak or no correlation between apoptosis and proliferation. However, a subset of aneuploid tumors had a high percentage of cells positive for Ki-67 and low percentage of apoptotic cells. Diploid tumors did not show any correlation between apoptosis and proliferation, although many of those tumors had both low apoptotic and proliferative indices. Whether those differences are of prognostic significance remains to be determined in follow-up studies that include more cases and clinical data. Here we have shown that LSC is a powerful new tool of potential clinical value for fast, objective analysis of apoptosis, proliferation and DNA ploidy in solid malignant tumors. PMID- 11064816 TI - Expression of vascular endothelial growth factor and basic fibroblast growth factor receptors in lung cancer. AB - OBJECTIVE: To determine the expression of two angiogenic factors, vascular endothelial growth factor (VEGF) and fibroblast growth factor receptors (FGFR), in non-small cell lung carcinoma (NSCLC) in relation to tumor stage (TN0, TN1, TN2) and in association with the expression of p53 protein, a potential suppressor of tumor angiogenesis. STUDY DESIGN: The immunohistochemical (IHC) expression of VEGF and FGFR was examined in paraffin sections of 56 NSCLC in relation to the presence of lymph node metastases and p53 expression. Nodal status of NSCLC determined: 27 tumors, N0; 16, N1; and 13, N2 stage. Semiquantitative analysis with a score corresponding to IHC staining intensity and percentage of positive cells was used. Statistical analysis was performed with the chi 2 test. RESULTS: A significant association was noted between VEGF and FGFR expression in NSCLC. No relation was found between VEGF, FGFR expression and lymph node metastasis or p53 expression. CONCLUSION: We assume that VEGF and FGFR act in a synergistic manner in NSCLC and that their expression is not related to lymph node metastases. Angiogenesis is a very complex phenomenon and heterogeneous within tumors. Also, it is affected by microenviromental factors. PMID- 11064817 TI - Transforming growth factor beta 1 and androgen receptors in prostate neoplasia. AB - OBJECTIVE: To investigate the interplay between transforming growth factor (TGF) beta 1, androgen receptors and stromal-epithelial interactions in benign prostatic hyperplasia (BPH), prostate intraepithelial neoplasia (PIN) and prostate carcinoma areas of prostate neoplasia. STUDY DESIGN: In this immunohistochemical study we investigated staining patterns and then determined the correlation between TGF-beta 1 expression and androgen receptor status in the epithelium and stroma of 60 paraffin-embedded tissues from radical prostatectomies. RESULTS: Staining patterns differed in the epithelium and stroma of tumor and peritumor prostatic tissue. TGF-beta 1 immunostaining (H-scores) in the epithelium and stroma increased significantly from BPH to PIN and from BPH to prostate carcinoma in the epithelium (P < .05), whereas androgen receptor (AR) immunoreactivity significantly (P < .05) increased from BPH to PIN to prostatic carcinoma in epithelium and stroma. TGF-beta 1 did not correlate with histologic grade of differentiation, whereas AR proteins were more strongly expressed in Gleason score 5 and 6 than score 7 tumors (P < .05). Nonlinear regression showed a significant correlation (P < .01) between TGF-beta 1 and AR expression only in the stromal compartment of PIN. CONCLUSION: These findings argue in favor of an interaction between TGF-beta 1 and AR in the early stages of prostate carcinogenesis and suggest that TGF-beta 1 plays a central role in stromal epithelial interactions during the early stages of malignant transformation. PMID- 11064818 TI - Superficial urothelial (umbrella) cells. A potential cause of abnormal DNA ploidy results in urine specimens. AB - OBJECTIVE: To determine the DNA ploidy distribution in urothelial superficial (umbrella) cells and to assess the value of the image analysis operator's experience. STUDY DESIGN: DNA ploidy was assessed in 12 cytologically negative bladder washes stained with Feulgen stain. All 12 cases were evaluated independently by three operators with different levels of cytopathology experience and different goals. Operator 1 (experienced) selected only nuclei of urothelial cells, avoiding nuclei of superficial cells; operator 2 (experienced) selected only nuclei of superficial cells; operator 3 (inexperienced) selected the largest and most-atypical-looking nuclei. Each operator measured a total of 100 nuclei per case. RESULTS: Operator 1 found all cases to be diploid (97% of nuclei on average). Operators 2 and 3 showed a wide range of results. Almost half the nuclei (47%) analyzed by operator 2 were in the diploid region, a third (35%) were in the tetraploid region, and the remaining (18%) ones had a DNA index (DI) in the range of 1.2-1.8 or > 2.5. Operator 3 obtained the most abnormal results. Only 9% of the nuclei were diploid, while 37% were in the tetraploid region, 18% were in the hyperploid region, and 35% had a DI in the range of 1.2-1.8. Differences among results obtained by each operator were statistically significant. CONCLUSION: The nuclei of superficial (umbrella) cells often have abnormal DNA content, which may cause abnormal DNA ploidy results in cytomorphologically normal bladder washes. Consequently, the nuclei of superficial cells should be avoided in the evaluation of urine samples. DNA analysis of urine specimens requires selection of nuclei only of deep urothelial cells by an experienced operator. PMID- 11064819 TI - Cytocentrifugation conditions affecting the differential cell count in bronchoalveolar lavage fluid. AB - OBJECTIVE: To investigate variations in speed, duration and acceleration rate of the Cytospin 3 cytocentrifuge (Shandon Scientific Ltd., Astmoor, England) on the differential cell count of bronchoalveolar lavage (BAL) fluid samples. STUDY DESIGN: BAL fluid samples (n = 51) were cytocentrifuged at various combinations of speed (500, 1,200 and 2,000 rpm), acceleration rate (low, medium and high) and duration (5, 10, 15 and 20 minutes). The preparations were May-Grunwald-Giemsa stained and differentiated on 500 cells. Data were analyzed by mixed model repeated measurements ANOVA. RESULTS: The mean lymphocyte count was significantly higher at 1,200 rpm than at 500 rpm, whereas the macrophage count decreased. Between 1,200 and 2,000 rpm, the number of both cell types stabilized. Significantly higher numbers of lymphocytes were recorded at 10 and 15 minutes of cytocentrifugation than at 5 minutes. The acceleration rate did not influence the differential cell count. Seventeen BAL fluid samples were selected to test the diagnostic impact of cell damage using a validated computer program. In 1 of 17 samples the predicted diagnosis did not correspond between two different speeds (500 and 2,000 rpm). CONCLUSION: Variations in cytocentrifugation speed and duration affected the mean lymphocyte and macrophage counts of BAL fluid samples. PMID- 11064820 TI - Automated image analysis for monitoring oxidative burst in macrophages. AB - OBJECTIVE: To evaluate oxidative bursts induced by phorbol myristate acetate in phagocytes at the single-cell level by automated image analysis. STUDY DESIGN: The generation of reactive oxygen species was quantitatively expressed by means of histograms displaying the percentage of cells corresponding to each of the total optical densities measured. RESULTS: Macrophage subpopulations were quantitatively defined. This method allows detailed analysis of the amount of formazan per cell and the sites of deposition of blue precipitate in each cell. CONCLUSION: Image analysis is a reliable quantitative, single-cell assay for studying various cellular characteristics associated with macrophage functions. PMID- 11064821 TI - Neurological syndromes associated with nervous system-specific autoantibodies. PMID- 11064822 TI - Monoclonal IgM autoantibody reactivity in M-IgM peripheral neuropathy. PMID- 11064823 TI - Paraneoplastic neurological syndromes associated with Yo, Hu, and Ri autoantibodies. PMID- 11064824 TI - Specificity of antiglycolipid antibodies. PMID- 11064825 TI - Relationship between autoantibody specificities and peripheral nervous system involvements. PMID- 11064826 TI - Anti-CV2 autoantibodies and paraneoplastic neurological syndromes. PMID- 11064827 TI - Immunological mechanisms in paraneoplastic peripheral neuropathy. PMID- 11064828 TI - Immunogenicity of glycolipids. PMID- 11064829 TI - Diabetic nephropathy in non-insulin dependent diabetes mellitus. Cardiovascular risk factors and antihypertensive treatment. PMID- 11064830 TI - Wine and mortality. Evidence for causal inference? PMID- 11064831 TI - Hemodilution and natriuresis of intravascular volume expansion in humans. PMID- 11064832 TI - Community-acquired bacteraemia and antibiotic resistance. Trends during a 17-year period in a Danish county. AB - INTRODUCTION: The aim was to ascertain the prevalence of antibiotic resistance among blood isolates from patients with community-acquired bacteraemia and to relate it to antibiotic consumption. METHODOLOGY: Cases of community-acquired bacteraemia were identified in a regional bacteraemia register in the County of Northern Jutland. The study included 3974 episodes in 3805 patients during a 17 year period. Total regional consumption of antibiotics was expressed in Defined Daily Doses (DDD). RESULTS: The prevalence of antibiotic resistance was stable with few exceptions. The most notable time trend was noted for Escherichia coli for which the prevalence of resistance to ampicillin increased from 17% (95% confidence limits (CL) 12-23%) to 28% (95% CL 23-33%); for other Enterobacteriaceae the increase was from 73% (95% CL 61-83%) to 86% (95% CL 77 92%). The prevalence of resistance to aminoglycosides, fluoroquinolones and third generation cephalosporins remained low among all isolates of Enterobacteriaceae. Regional antibiotic consumption ranged from 10.2 to 13.6 DDD/1000 inhabitants/day. Consumption of penicillins with Gram-negative spectrum reached a maximum of 4.6 DDD/1000 inhabitants/day in 1993 and decreased towards the end of the study period. The prevalence of ampicillin-resistant E. coli was positively correlated with consumption of penicillins with Gram-negative spectrum; the correlation was stronger when adjustment was made for co-selection by tetracyclines and sulphonamides. CONCLUSION: Therapeutic options for community acquired bacteraemia have not yet become seriously limited by prevalence of acquired antibiotic resistance. Still we found some evidence that consumption of penicillins with Gram-negative spectrum, sulphonamides and tetracyclines promotes antibiotic resistance among Enterobacteriaceae. PMID- 11064833 TI - The prevalence of obsessive compulsive symptoms in a sample of Egyptian psychiatric patients. AB - Obsessions can occur in many psychiatric disorders or they may constitute the entire illness, which is then referred to as an obsessional state (Rees, 1993). The relationship of obsessive compulsive symptoms (OCS) to different psychiatric disorders is still controversial. This work was undertaken to study the co occurrence and phenomenology of OCS with other psychiatric disorders. We examined a sample of 372 psychiatric outpatients using the arabic version of Yale Brown obsessive-compulsive symptom (Y-BOCS) checklist and compared them with a control group composed of 308 non-psychiatric subjects. Subjects were additionally assessed by means of the obsession symptom section of the PSE (10th) edition for trait rating, the arabic version of the Eysenck rigidity scale and the arabic version of the religious orientation scale. OCS were found to be significantly higher in the different psychiatric categories than in the non-psychiatric categories; 83% of patients with neurotic, stress related and somatoform disorders, 51% of patients with mood disorders and 47% of patients with schizophrenia, schizotypal and delusional disorders were found to have OCS in their symptomatology. Furthermore, the data suggest that OCS in psychiatric patients have a distinct phenomenology from that in non-psychiatric subjects. The results did not however reveal a relationship between OCS and either rigidity or religious orientation. PMID- 11064834 TI - [Evaluation of the quality of life of schizophrenic patients: validation of the brief version of the Quality of Life Interview]. AB - The main objective of the study was to determine the psychometric properties (mainly, validity and reliability) of the French language version of the brief Quality of Life Interview (QoLI). That instrument evaluates both the subjective and objective aspects of quality of life. METHODS: 128 patients fulfilling the DSM IV criteria for schizophrenia were included. Quality of life was evaluated using the brief QoLI. Schizophrenic symptoms were evaluated using the Positive and Negative Syndrome Scale (PANSS). The validity of the internal structure of the QoLI was investigated by means of item analysis, study of the correlations between the items and between the item-dimensions and principal component analysis addressing the subjective fields. The validity of the external structure was mainly investigated through the nomological validity study. The reliability of the scale was evaluated by studying the internal consistency. In addition, the acceptability of the scale was documented. RESULTS: The results of the study of the validity of the internal structure confirmed the pertinence of the pre defined fields, particularly the subjective fields. Only some of the subjective fields of the QoLI showed significant correlations with the PANSS sub-scales. The acceptability of the scale was satisfactory. CONCLUSIONS: The psychometric properties of the French language version of the brief QoLI appeared satisfactory. The brief QoLi was shown to be an easily used instrument for assessment of the various objective and subjective aspects of the quality of life of schizophrenic patients. PMID- 11064835 TI - [Polysomnographic aspects of antidepressive therapy]. AB - Sleep disorders are very common in depression. They have been quantitatively and qualitatively described by polysomnographical recordings. It is of interest to know how treatments act on polysomnographical data. In this article, we propose to evaluate five treatment approaches proposed for depressive illness. Pharmacological treatment induces marked changes in sleep continuity, sleep architecture and REM sleep. However, no specific sleep profiles emerge neither for each treatment class nor for molecules within the same pharmacological class. But actually, sleep data cannot be viewed as markers of treatment response. Psychotherapeutic interventions have only few effects on sleep of depressed patients. Treatment efficiency does not seem to be correlated to abnormal sleep parameters. Sleep deprivation induces marked changes in nearly all sleep parameters and in temporal distribution of sleep stages during the night. Actually, the efficiency of sleep deprivation cannot longer be explained by suppression of REM sleep. Sleep deprivation has only a transient effect and treatment indications are therefore secondary. Sleep parameters do not distinguish responders from non-responders. Sleep deprivation shows that there is a depressogenic effect of sleep in the end of the night. Bright light therapy shows marked changes in sleep continuity parameters. Among all studies that examine the impact of treatment on sleep EEG, ECT has received little attention. The few studies available are either case studies or with poor effectifes++. For this reason and because of methodological bias, results are heterogeneous and no definite conclusions can be drawn. But all of them agree that ECT modifies sleep EEG. So, changes in polysomnographical data cannot predict response to any treatment. Prospective sleep studies are difficult to realise on a great number of patients explaining absence of treatment predictors. PMID- 11064836 TI - [Fratricide occurring during a nocturnal episode of somnambulism: a case report]. AB - Occasionally, some people developed violent acts during their sleeping. Fortunately, these acts remain virtual and do not have serious life consequences. However, the execution of the dramatical act may happen in setting automatic activities. The manifestations of these acts are exceptional but very often misappreciated. Among them, we have been confronted to a case of "fratricide expertise" in the north of France. The person concerned is a teenager of 17, accused of the murder of his young brother, by several stabs, in the night. This homicide could be the consequence of multitude of factors. Though he hasn't shown clinical somnambulism, the results of electroencephalogram tracing were symptoms of predisposing factors. We can also imagine the facilitating effect of other factors, ephedrine, more particularly. The rapport with the relevant documents follows an expert's report and can be added to the cases published in the literature in which it was shown that the automatic activity may be at the origin of the medico-legal act of somnambulistic patients. Finally, we have been able to deduce two classes of factors, which may be responsible for violent act execution. PMID- 11064837 TI - [Critical review of measures of quality of life in schizophrenia]. AB - In order to provide clinicians, researchers, program evaluators and administrators with current information on the assessment of humanistic outcomes of services for schizophrenic patients, a literature review is performed in which references to quality of life (QOL) assessment were made in the context of schizophrenia. Measures are summarized according to purpose, content, psychometric properties. Fifteen QOL instruments are summarized and reflect considerable variability on the relevant criteria: 11 are developed for persons with severe and persistent mental illnesses and used among populations including a major part of schizophrenic patients; 3 are specific QOL measures for schizophrenic patients; 1 is a generic QOL instrument used among psychotic patients. Given that none of these QOL measures has been widely used or accepted as a standard, the choice of a measure must rest on the investigator's particular purpose and needs. CONCLUSION: There is a clear need for a QOL instrument that is specific for schizophrenia, given its high prevalence and chronic nature. The lack of information related to responsiveness of these scales stresses the problems of their inclusion in clinical trials. Scales that have been used in studies of schizophrenia were nearly all developed in the United States and the relevance of their content must be questioned. Rather than relying on the literature or experts to determine those needs that are important to patients with schizophrenia, the content of the instrument should be derived from qualitative interviews with patients who are at different stages of their illness. Moreover, the questionnaire should be self-administered. PMID- 11064838 TI - [Validation of French versions of magical ideation and perceptual aberrations questionnaires]. AB - Chapman and colleagues have developed symptom-oriented scales based on Meehl's manual of schizotypy, such as the Social Anhedonia (SA) and Physical Anhedonia (PhA) Scales, the Magical Ideation Scale (MIS), and the Perceptual Aberration Scale (PAS). Whereas Chapman's scales of psychosis proneness are the most internationally used instruments for the assessment of schizotypy, some of them, such as MIS and PAS, were still not available in French. We reported here the validation study of the MIS and the PAS French versions that we had published previously. This study was conducted in a sample of 233 students (males: n = 108; females: n = 125; mean age: 21.17 +/- 1.47; mean educational level: 13.36 +/- 1.06). The French versions of the MIS and the PAS have high internal reliability (MIS: Cronbach's alpha = 0.85; PAS: Cronbach's alpha = 0.88). French norms are given for each of these scales. They are respectively 19/30 for the MIS and 17/35 for the PAS high cutoff scores without any difference when gender was considered. These results are very closed to those found by Chapman and colleagues for University of Wisconsin undergraduate students. PMID- 11064839 TI - [Epidemiological study of cannabis abuse and dependence in 256 adolescents]. AB - A study of cannabis use in French adolescents was implemented using a questionnaire derived from the French version of the Mini International Psychiatric Interview based on DSM IV criteria for cannabis dependence, among a population of 256 high school students from two high schools with different SES backgrounds. Among the adolescents studied, 41.4% (n = 106) reported using cannabis occasionally or regularly, 51.2% had never used cannabis, and 7.4% had used cannabis and subsequently quit. Incidence of usage appeared to increase as a function of age, 51.4% for those 18 years and over, 44.6% for 17 years-old, and 30.4% for 16 years-old, girls tending to consume less than boys (36.4% to 45.2%). With regards to results of the MINI, of the regular or occasional users (n = 106), 47.2% of the subjects indicated substance dependence while the remaining subjects indicated that they were recreational users only. Among users, data concerning tolerance, withdrawal, and excessive consumption indicated that subjects were significantly affected by their addictive behavior; 33% of users reported having smoked cannabis for one year or less with 10.4% reporting that they have smoked for more than three years. Among those having smoked one year or less, 31.4% reported signs of dependence versus 68.6% who consume on a recreational basis; among those having used cannabis for three years or more, 63.6% reported dependence while 36.4% admitted to recreational usage. This study indicates the seriousness of cannabis usage among high-school students, underlining addictive and dependent behavior and their effects on daily life as well as significant the increase of usage with age and as a function of number of years of smoking. PMID- 11064840 TI - [Apropos of sleep quality in students: prospective study]. AB - This study about the pupils' sleep in the academy of Aix-Marseille has been realized after the observations of second cycle's teachers; they have noted a somnolence by an important number of pupils during lessons, consequence of a supposed sleep lack able to have repercussions on the scholastic success. Hearing teachers and pupils' suggestions, the authors have established a questionnaire with 20 items; this study is concerning 1,300 pupils, 15 to 20 years old, in classic, technical and agricultural high schools, both in public and private establishments. The most salient characteristic is the differences between the sleep of the scholastic period and the sleep of week end or holiday. In the future, a better knowledge of pupils' sleeping habits could be the starting point of information and prevention campaign. PMID- 11064841 TI - [Sulpiride: study of 669 patient presenting with pain of psychological origin]. AB - Among somatoform disorders, pain disorder (DSM IV) appears to be relatively common in general practice and to cause social, psychological, and functional impairment. A previous study conducted by Lemoine (1997) has shown that sulpiride is more effective than placebo in reducing intensity and frequency of pain in this disorder. The aim of our study was to assess safety and efficacy of sulpiride in a large sample of patients under natural conditions of use, in general practice. In a multicenter, open clinical trial, 669 patients (mean age: 47 years +/- 12; male: 245, female: 424) fulfilling the DSM IV criteria for pain disorder (of gastrointestinal localization), were included by 321 general practitioners (GP) and treated for 6 weeks with sulpiride 150 mg/d. Investigators' evaluations were planned at D14 and D42. Furthermore a diary was given to each patient for self evaluation and intercurrent events reporting. The pain was of psychological type in 93% of cases and caused social or working disabilities in 78% of patients. At inclusion the mean score of the Hamilton Anxiety Rating Scale was 18 +/- 8, and the mean score of the depression scale HARD (Humeur, Angoisse, Ralentissement, Danger) was 14.8 +/- 6.4. During the study 7.9% of the patients had at least one adverse event, and 3% of patients were withdrawn for adverse event. Safety assessed with a specific variable (grouping together adverse events' reporting and results of CGI item 3) was good for 88% of patients. The principal criterion of efficacy was the clinician's evaluation of the intensity and frequency of abdominal pain on a four-point scale from 0 (asymptomatic) to 3 (important/continuous) from D0 to D End a decrease in pain intensity (91% of patients) and in pain frequency (89%) was observed as well as in frequency and intensity of related gastroenterological symptoms such as disturbances of bowel movements (79% and 78%), bloated symptoms (88% and 83%), nausea/vomiting (90% and 90%). A similar improvement (p < 0.001) was observed from D0 to End point on the self evaluation parameters (Visual Analogic Scales), assessing pain (mean score D0-D End: 17.1 +/- 15.9), quality of sleep (mean score D0-D End: 27.1 +/- 17.8), activity (mean score D0-D End: 24.4 +/- 18.8), and appetite (mean score D0-D End: 22.6 +/- 16.6). In conclusion these results confirm the usefulness of sulpiride in the treatment of pain disorders a symptomatology known to cause difficulties to GP's in their practice. PMID- 11064842 TI - [Neuropsychiatric symptoms in preventive antimalarial treatment with mefloquine: apropos of 2 cases]. AB - Two observations of severe neuropsychiatric reactions occurring during chemoprophylaxis with mefloquine are reported. The first case regards a 43 years old woman who developed a severe depression with visual and auditive hallucinations and a paranoid delusion. She was treated by clomipramine and risperidone. The second case concerns a 55 years old man who developed an acute psychosis with confusion. He was treated with halopridol during a short time. He presented twice an acute psychosis during a chemoprophylaxis with mefloquine. Several cases of neuropsychiatric side effects with mefloquine chemoprophylaxis or treatment have been described. Authors estimate that one of 250 therapeutic users has severe neuropsychiatric reactions, compared with one of 10,000 to 15,000 in the prophylaxis users. Disorders could last from 15 minutes to several weeks. Women and patients with personal or familial antecedents of psychiatric disorders are more frequently concerned. Alcohol and the association with other antimalarial drugs (like quinine) are two other risk factors. Therefore, some advices may be suggested regarding the use of mefloquine for malaria prophylaxis and treatment. PMID- 11064843 TI - [Klein-Levin syndrome: a neurological disease with psychiatric symptoms]. AB - Kleine-Levin syndrome belongs to the recurrent hypersomnias group. It is a rare and benignant disease, occurring in young men 10 to 25 years old. The diagnosis is first clinical and the hypersomniac episodes, joined to psychiatric symptoms, are irregularly recurrent during few years. Diagnosis is uneasy during the first episode and in the attenuated forms... 500 cases have been described all around the world but it's highly likely that many patients haven't been listed. This syndrome, just like Gelineau disease, stands in the group of primary pathological hypersomnias. In a clinical point of view, the cardinal and constant symptom is hypersomnia. Psychiatric symptoms can be irregularly joined: megaphagia, sexual behavioural disorders, thymic disorders, personality modifications. The clinical examination is poor and aspecific. During an hypersomniac episode, a polygraphic recording during 24 or 48 hours will give diagnosis informations (fragmented and unstable sleep, reduction in stages 3 and 4 of non-REM sleep, reduction in REM sleep latency) and a biological and radiological evaluation will be necessary to exclude organic etiology (tumoral progres, infectious disease...). In a therapeutic point of view, prescription of psychostimulant drugs is recommended during fits and some treatments are used in a preventive way (lithium and carbamazepine). PMID- 11064844 TI - [Natural evolution of paranoid schizophrenia treated consecutively with 4 different neuroleptics]. AB - Ms M is a young female patient (born 1968) with paranoid schizophrenia. The first acute psychotic episode, which involved marked delirium, was treated with an atypical antipsychotic agent. During the next year, renewed hospitalisation and treatment with two other antipsychotics resulted only in partial improvement, and troublesome adverse effects were noted. Amelioration of all symptoms, both psychotic and debilitating, was finally obtained through prescription of a third atypical antipsychotic with which the patient was highly satisfied. We examine the value of first-line treatment of schizophrenia with atypical antipsychotics as of the first acute episodes of acute delirium in the light of current data in the literature. PMID- 11064845 TI - [The Defense Style Questionnaire of 40 items: disappointing results of a first French validation study]. PMID- 11064846 TI - Advocacy is needed to promote research into diseases of physical inactivity. PMID- 11064847 TI - Muscle glucose transporter (GLUT 4) gene expression during exercise. AB - Among the many effects of exercise is the induction of glucose transporter (GLUT 4) expression in skeletal muscle. In this review, we examine the intracellular signals that may mediate the exercise-induced increase in GLUT4 gene transcription. This induction is likely to be dependent upon intracellular calcium concentrations and the energy charge of the muscle. PMID- 11064848 TI - Environmental determinants of physical activity and sedentary behavior. AB - Environmental changes are expected to lead to decreased time in sedentary behavior and to increased levels of physical activity in populations. Past research has emphasized psychosocial determinants of physical activity. Progress in the field will require more focus on understanding sedentary behaviors and the role of environmental determinants. PMID- 11064849 TI - The Na+/K(+)-pump protects muscle excitability and contractility during exercise. AB - In skeletal muscle, the concentration of Na+/K(+)-pumps is high and increases through training. In isolated muscles, contractile endurance depends in part on Na+/K(+)-pump concentration. Exercise leads to rundown of Na+/K(+)-gradients, compound action potentials, and force. Early and efficient activation of the Na+/K(+)-pump, however, protects excitability and contractility. PMID- 11064850 TI - Exercise alone is an effective strategy for reducing obesity and related comorbidities. AB - The commonly held view that exercise alone is not a useful strategy for obesity reduction is drawn from studies with limitations that confound interpretation. Recent evidence counters the dogma that daily exercise produces only modest weight loss and suggests that exercise without diet restriction is an effective strategy for reducing obesity and related co-morbidities. PMID- 11064851 TI - Does muscle function and metabolism affect exercise performance in the heat? AB - It has been suggested that exercise performance in the heat is limited by the degree of hyperthermia, which, in some circumstances, compromises cardiovascular function and/or the central nervous system. However, this review presents evidence that a temperature-induced dysfunction to skeletal muscle contraction may contribute to a reduction in performance during exercise in the heat. PMID- 11064852 TI - Chronic neural adaptations to unilateral exercise: mechanisms of cross education. AB - Cross education refers to the contralateral effect of chronic motor activity in one limb. The effect can enhance or diminish motor activity and is specific to the homologous muscles and the training task. The mechanisms underlying the phenomenon involve adaptations in the nervous system, probably at the level of the spinal cord. PMID- 11064853 TI - Role of nitric oxide and prostaglandins in the bone formation response to mechanical loading. AB - Nitric oxide and prostaglandins are crucial early mediators in mechanically induced bone formation. They are also responsible for the associated induction of gene expression of c-fos and IGF-1 in osteocytes, key mechanosensory cells in bone. Insight into the cellular and molecular mechanisms underlying bone formation has important implications for the maintenance of structural competence of bone. PMID- 11064854 TI - Antioxidants and the eye. PMID- 11064855 TI - Nutritional influences on risk for cataract. PMID- 11064856 TI - The effect of ascorbate on wound healing. PMID- 11064857 TI - Vitamins C and E in cataract risk reduction. PMID- 11064858 TI - Antioxidants and cataract formation: a summary review. PMID- 11064859 TI - Vitamin A deficiency and the eye. PMID- 11064860 TI - Nutrition and retinal degenerations. AB - Considerable progress has been made in the understanding and management of degenerative diseases of the retina involving photoreceptors. Nutritional approaches to treatment have proved successful in the case of the common forms of retinitis pigmentosa (supplementation with vitamin A), Bassen-Kornzweig disease (supplementation with vitamins A, E, and K), gyrate atrophy (low-protein, low arginine diet and/or supplementation with vitamin B6), and Refsum disease (low phytol, low-phytanic acid diet). The night blindness associated with Sorsby fundus dystrophy can be reversed over the short term with vitamin A. A significant trend for decreased risk for advanced or exudative ARMD has been reported among those whose diets contain a higher content of carotenoids, such as spinach and collard greens. A randomized trial is in progress to determine whether beta-carotene, vitamin C, and vitamin E as well as trace minerals, particularly zinc, will modify the course of ARMD. The difficulties that patients with retinal degenerations face as a result of their diminishing vision, sometimes over decades, cannot be underestimated. Nutritional therapy has proved effective in modifying the course of a number of these conditions; the therapeutic benefit of nutritional modification in diseases that have a genetic basis is of particular interest. Further research is warranted to determine the mechanisms by which these treatments provide their benefit as well as to identify other conditions that may yield to nutritional intervention. Risk-factor analyses of well-defined populations followed over time with food-frequency questionnaires in conjunction with careful assessments of visual function may reveal other dietary constituents that can modify the course of degenerative diseases of the retina. PMID- 11064861 TI - Serum application for the treatment of ocular surface disorders. PMID- 11064862 TI - [Hyperbaric oxygenation and neurosurgery]. PMID- 11064863 TI - [Radiological aspect of developmental neuroanatomy: forebrain embryogenesis and its disorders]. PMID- 11064864 TI - [Clinical problems in the treatment of primary central nervous system malignant lymphoma]. AB - While the treatment of extracerebral lymphomas with the combination of chemotherapy and radiotherapy has drastically improved outcomes, the treatment of primary central nervous system malignant lymphoma(PCNSML) has been disappointing. Because of the variety of clinical presentations, progression of disease, and treatment modalities, careful inspection of factors which influence survival may suggest possible approaches for a more effective management in each case. In this report, clinical problems in the treatment of PCNSML was discussed with presenting cases experienced at Tenri hospital since 1983. There were twenty-one cases of histologically proven PCNSML. Surgical resection was undertaken in 11 patients. The remaining patients underwent biopsy only. All patients received radiation therapy. Various modality of chemotherapy was performed in 14 cases. Tumor recurrence was occurred at one or more CNS sites, including 3 patients who had meningeal relapse and one patient who also relapsed outside the CNS. The median survival time was estimated to be 15 months. Among the 9 patients alive at last contact the median length of follow up was 31.5 months(range 4 to 56 months). The following problems were presented and discussed; 1) varieties of initial clinical presentations, 2) variable recurrence patterns, metastases including CSF dissemination and extracranial metastases, 3) treatment-related leukoencephalopathy. PMID- 11064865 TI - [Diagnosis of acute cerebral infarction using diffusion-weighted imaging by low field (0.2 T) magnetic resonance image]. AB - The purpose of this study is to confirm the diagnosis of acute cerebral infarction on diffusion-weighted imaging using low field (0.2 T) magnetic resonance image(MRI). Acute cerebral infarctions in 51 patients were examined on diffusion-weighted imaging using low field MRI within 48 hours after clinical symptoms. Diffusion-weighted imaging was examined using line scan method. Twenty four cases were cortical infarction, and twenty-two cases were perforating infarction. In five cases out of 51 cases, ischemic regions were not detected as abnormal high signal intensity area on diffusion-weighted imaging. Four cases of no abnormal detection were transient ischemic attack, and the other one was a perforating infarction. The earliest detection time in cortical infarction cases was 1 hour and 20 minutes. On the other hand, the earliest detection time in perforating infarction cases was 3 hours. Detective ability for acute cerebral infarction on diffusion-weighted imaging by low field MRI was depending on both size and lesion of infarction. That is to say, either small size or brain stem infarction was hard to detect. Thin slice and vertical slice examination for the infarction may improve to diagnose in low field MRI. Our conclusion is acute cerebral infarction was able to be diagnosed on diffusion-weighted imaging by low field as well as high field MRI. PMID- 11064866 TI - [Neonatal hydrocephalus-volume determinations using computed tomography]. AB - Computed tomography potentially offers the most accurate noninvasive means of estimating in vivo volumes. Contiguous 5 and 10-mm-thick CT scans were obtained through phantom and neonatal cranium. Cross-sectional areas were calculated for each individual scan and volumes then determined with summation-of-areas technique. The indirect intracranial volume measurement technique was then used to compare hydrocephalic and non-hydrocephalic intracranial and ventricular volumes in nine neonates. Our findings show that intracranial volumes of hydrocephalic neonates with head circumference of more than 39 cm, have 1.97 times larger than control group. The ratios of lateral ventricle versus intracranial volume are 0.57 +/- 0.2 in hydrocephalic babies and 0.0062 +/- 0.001 in control babies, respectively(p < 0.005). PMID- 11064867 TI - [Spinal cord infarction presenting Brown-Sequard syndrome with impaired position sense]. AB - A 79-year-old hypertensive man presented left hemiplegia of sudden onset. Neurological examination revealed weakness of the left extremities, with hypoalgesia on the opposite side below the level C 4. He also showed Horner syndrome, facial hypoalgesia, weakness of sternocleidmastoid and trapezius muscles on the paralyzed side. The position and vibration senses were impaired on the left extremities. The position sense was more disturbed on his upper limb, to the extent that the patient was not able to recognize where his wrist was located. The cervical MRI exhibited a high signal intensity on the left half of the cord between C 2-C 5 vertebral level on T 2 WI. Bilateral vertebral arteries were patent, though severe stenosis of internal-external carotid artery bifurcation was observed on MRA. Asymmetrical distribution of upper cervical cord arteries, severe atherosclerotic change of cervical and intracranial vessels, and spondylotic cervical canal stenosis were suggested to contribute to cause the lateralized infarction of the cord, involving not only the anterior, but also posterior part, where Burdach's fascicle were probably more affected. PMID- 11064868 TI - [Neurosarcoidosis with girdle sensation and polyradiculoneuropathy masquerading as Guillain-Barre syndrome]. AB - We herein report two patients with neurosarcoidosis presenting girdle sensation in the trunk and polyradiculoneuritis. The first patient, a 53-year-old woman, manifested subacute progressive paresthesia in all four limbs and below the Th 3 level with girdle sensation from the thorax to lower abdomen and mild weakness in the left upper limb and the bilateral lower limbs. The patient was diagnosed to have sarcoidosis based on a biopsy of the scalenus anticus lymph nodes. The second patient, a 63-year-old woman, showed an acute onset of weakness and paresthesia in all four limbs and girdle sensation from the Th 5 to Th 8 level. On examination, she demonstrated diminished tendon reflexes in all four limbs, mild to severe weakness in all four limbs, paresthesia in all four limbs and below the Th 5 level. Although Guillain-Barre syndrome was initially suspected in this patient, the presence of girdle sensation led us to examine the possibility of neurosarcoidosis. Her examination demonstrated an abnormal accumulation of gallium in the bilateral hilar lymph nodes and mediastinum on scintigraphy, an elevated CD 4/CD 8 ratio in the bronchoalveolar lavage fluid, a negative tuberculin reaction, and elevated serum lysozyme level. These findings thus fulfilled the clinical criteria for sarcoidosis. None of the two patients showed any abnormalities in the thoracic cord MRI. In the first patient F wave was not evoked in either upper or lower limbs, while in the second patient temporal dispersion on M wave was observed in the right median and both ulnar nerves. We therefore consider the girdle sensation to have not been caused by myelopathy but instead by polyradiculopathy. When sarcoid peripheral neuropathy masquerades as Guillain-Barre syndrome, then the presence of girdle sensation may help diagnosis of neurosarcoidosis. PMID- 11064869 TI - [A case of olfactory neuroblastoma with intracranial extension and distant metastasis]. AB - Olfactory neuroblastoma is a rare tumor originating in the upper nasal cavity. It rarely extends intracranially. We report a clinical case of olfactory neuroblastoma with intracranial extension and distant metastasis. A 35-year-old man complained of nasal stuffiness and bleeding, headache and vomiting. Neurological examination showed anosmia and papilledema. MRI showed a huge mass that occupied the right nasal and paranasal cavities, and extended into the right frontal base. The tumor was removed totally and was histologically diagnosed as olfactory neuroblastoma. About two months after surgery, however, MRI demonstrated a rapid recurrence of the tumor in the nasal and paranasal cavities and the frontal lobe. Metastatic lesions were also seen in the right cervical lymph nodes. Chemotherapy was administered using cisplatin and etoposide. The tumor in the frontal lobe shrunk markedly but the other lesion persisted. Whole brain irradiation and local irradiation of the cervical lymph nodes were performed. The tumors became smaller but did not disappear. MRI demonstrated spinal dissemination. Irradiation of the whole spinal cord was performed. The metastatic lesions disappeared. The patient was discharged without neurological deficits, but died of pneumonia 15 months after surgery. Olfactory neuroblastoma is a slow-growing tumor and is highly radiosensitive, but it rarely extends or develops multiple distant metastases and seldom shows a short survival time, as in our case. A review of the literature documented responses in patients treated with a cisplatin-based drug combination. We recommend systemic control using cisplatin-based chemotherapy in addition to irradiation to prevent local recurrence in cases of advanced or metastatic olfactory neuroblastoma. PMID- 11064870 TI - [A case of intracerebral and subarachnoid hemorrhage of unknown origin with preceding headache]. AB - We report a case of a 31-year-old female with multiple intracerebral hemorrhage and subarachnoid hemorrhage. She presented with headache one week before hemorrhage, and a CT scan performed at that time showed no abnormal findings. Neurological examination on admission revealed mild disturbance of consciousness, papilledema, and mild left hemiparesis. CT scans demonstrated intracerebral hemorrhage in the right caudate head and left frontal subcortex, and diffuse subarachnoid hemorrhage. Cerebral angiogram and laboratory examination revealed no abnormal findings. Erythrocyte sedimentation rate, C reactive protein and antiphospholipid antibody were within normal ranges. The patient underwent removal of hematoma by craniotomy. One week after the operation, a subcutaneous hematoma in the area of the craniotomy was found. Cerebral angiography demonstrated an aneurysm of the right superficial temporal artery, which was remote from the craniotomy. This aneurysm was surgically removed and examined. Histopathological examination revealed the presence of a pseudoaneurysm but no inflammation. Although primary angitis of the central nervous system was suspected to be the cause of this disease, a definite diagnosis could not be obtained. PMID- 11064871 TI - [A case of giant skull myofibroma occupying left anterior cranial fossa]. AB - We report a rare case of giant skull myofibroma occupying left anterior cranial fossa. A 53-year-old woman presented with left exophthalmos for 2 years. Neurological examination showed left exophthalmos, disturbance of bilateral visual acuity, and bitemporal hemianopsia. A CT scan revealed an ossifing mass at left anterior cranial fossa. On magnetic resonance images, the tumor showed iso intensity on T 1-weighted image, heterogeneous high intensity on T 2-weighted image, and was heterogeneously well-enhanced after administration of Gd-DTPA. The tumor was fed mainly by middle meningeal artery. The patient underwent surgery and the tumor was removed totally. Histological diagnosis of the tumor was myofibroma. The patient has been followed every other month by MRI without any adjuvant therapy. There has been no tumor recurrence for 19 months. There is no other myofibroma in her body, therefore the patient was diagnosed as solitary myofibroma of the skull. Our case is the first report of solitary myofibroma of the skull because we could not find any reports on solitary myofibroma of the skull in the past literature. PMID- 11064872 TI - [A case of delayed subrachnoid hemorrhage from vertebrobasilar artery dissecting aneurysm]. AB - We report a case of delayed subarachnoid hemorrhage (SAH) from a vertebrobasilar artery dissecting aneurysm (VBA-DA). The patient was a healthy 32-year-old woman with a sudden onset of severe occipitalgia. Next day, her headache improved gradually, and she consulted with our department. Although we initially suspected that she was suffering from SAH, neurological findings, CT, and cerebrospinal fluid examination did not reveal any abnormal conditions, including SAH. Therefore, she was treated conservatively with analgesics. Twelve days after the initial onset of the headache, she was admitted because of severe re-attack of headache, rt. hemiparesis with rt. oculomotor nerve palsy and loss of consciousness. CT revealed moderate SAH and cerebral angiograms showed VBA-DA. After the cerebral angiography, bleeding reoccurred two times and she lost her life. We present the case, review the literature and discuss the relationship between presenting symptom of headache and non-hemorrhagic VBA-DA. A few cases of non-hemorrhagic VBA-DA have been reported in the literature in which the only presenting symptom was headache, followed by delayed SAH from non-hemorrhagic dissecting aneurysm. Consequently, we concluded that her initial symptom of headache was due to dissection of vertebrobasilar artery, and that SAH was due to delayed hemorrhage of non-hemorrhagic VBA-DA. Even when neurological findings, CT and cerebrospinal fluid examination reveal no abnormalities in the early stage after the sudden onset of headache, especially in the occiptal or nuchal regions, non-hemorrhagic VBA-DA, which has a risk of fatal hemorrhage, cannot be ruled out with certainty. Therefore, MRI, MRA, three-dimensional CT, or cerebral angiography should be performed in such cases. PMID- 11064873 TI - [Acute epidural hematoma caused by contrecoup injury]. AB - We report a rare case of the acute epidural hematoma caused by contrecoup injury. A 59-year-old woman came to our hospital on foot because of her head injury. She slipped on a stone and hit her occiput against the concrete floor without loss of consciousness. On admission she was alert and complained of dizziness and nausea without neurological deficits. She had a subcutaneous hematoma in the occiput. There was no Battle's sign nor cerebrospinal fluid leakage. Skull film revealed a linear fracture of the occipital bone. Computed Tomography(CT) of the brain showed the acute epidural hematoma at the left frontal region. She was treated conservatively and discharged 19 days after injury without neurological deficits. The mechanism of this lesion is speculated as follows: the deformation of the skull and the negative pressure produced in the frontal region with the occipital injury stripped the dura mater from the calvarium, which lead to the rupture and hemorrhage of the small interposed vessels resulting in the epidural hematoma. PMID- 11064874 TI - [Intraventricular hemorrhage in the mature infant]. PMID- 11064875 TI - [Diffusion weighted MR imaging of Wernicke encephalopathy]. PMID- 11064876 TI - From school paper to published article. PMID- 11064877 TI - The efficacy and cost effectiveness of new antiemetic guidelines. AB - PURPOSE/OBJECTIVES: To develop antiemetic guidelines to improve efficacy, optimize nursing and pharmacy time, increase compliance, and enhance cost savings. DESIGN: Prospective, descriptive survey. SETTING: Outpatient oncology clinic in a large metropolitan city in the mid-Atlantic United States. SAMPLE: 90 patients were evaluated for the study; 52 patients met the eligibility criteria. METHODS: A standard antiemetic form was developed containing the emetogenic classification of the chemotherapeutic drugs and the recommended antiemetic regimen. A patient diary and visual analog scale measured patient satisfaction with the regimen-measured outcomes. MAIN RESEARCH VARIABLES: Episodes of nausea and vomiting, classification of chemotherapeutic drugs, and patient satisfaction. FINDINGS: Seventy-nine percent of patients receiving highly emetogenic chemotherapy demonstrated complete protection from nausea and vomiting during the first 24 hours post-therapy. All the patients receiving moderately and mildly emetogenic regimens achieved complete protection. Patients who received cisplatin containing regimens and were on the delayed regimen of antiemetics demonstrated complete protection on days two through seven. The mean overall score for patient satisfaction with the regimen was 89 mm on a 100 mm visual analog scale. CONCLUSIONS: The new oral antiemetic regimen compared favorably with published data, was well-tolerated, and resulted in lower pharmacy and nursing costs, with a cost saving potential of $20,000 per year. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses must be able to implement state-of-the-art knowledge of chemotherapy, antiemetics, and nonpharmacologic interventions to effectively manage the care of patients receiving chemotherapy. This must be performed to achieve cost effectiveness as well as useful clinical outcomes. PMID- 11064878 TI - A narrative study of chemotherapy-induced alopecia. AB - PURPOSE/OBJECTIVES: To describe the experience of alopecia in people undergoing chemotherapy. DESIGN: Qualitative. SAMPLE: Using announcement flyers, 15 participants (13 women and 2 men) were recruited to participate in audiotaped, in depth interviews. METHOD: In-depth interviews and narrative analysis of participants' "stories" using a sociolinguistic approach to narrative analysis. FINDINGS: Alopecia is a significant and disturbing side effect of chemotherapy. Preparing for hair loss, experiencing hair falling out, realizing an altered sense of self, trying to look normal, being reminded of disease, joking about alopecia, sharing being bald, having problems with wigs, taking control, and experiencing hair growing back emerged as aspects of the experience. CONCLUSIONS: Understanding the full significance of the experience of alopecia in an individual's everyday life and personal identity is critical to providing support during the course of illness and developing strategies to help clients cope with the difficult changes that occur during cancer treatment. IMPLICATIONS FOR NURSING PRACTICE: Information about alopecia can help to cognitively prepare the person, but the emotional response to alopecia is difficult to anticipate. Nurses need to create an atmosphere that encourages patients to tell their stories. PMID- 11064879 TI - Testing the effects of a decision-making and risk-reduction program for cancer surviving adolescents. AB - PURPOSE/OBJECTIVES: To test the effects of a decision-making and risk-reduction program for cancer-surviving adolescents. DESIGN: Prospective clinical trial using a quasi-experimental pretest/post-test design with repeated measures. SETTING: Two survivor follow-up clinics and a camp for children and adolescents with cancer located in upstate New York. SAMPLE: A convenience sample of 64 survivors (13-21 years of age). The intervention group consisted of 21 survivors who attended a workshop, and the comparison group consisted of 43 survivors who did not attend the workshop. METHODS: Intervention-integrated information specific to survivorship, decision-making skills, risk behaviors, and social support from peers and healthcare professionals. The educational component of the program lasted one day (five one-hour units), and the social component lasted overnight. A single, semistructured interview at the time of the previous yearly evaluation visit was used for baseline data. Testing was conducted during home visits at 1, 6, and 12 months. MAIN RESEARCH VARIABLES: Decision making, risk motivation, and risk behaviors (i.e., smoking, alcohol use, and illicit drug use). FINDINGS: The effect of the intervention for improving decision making was significant at 1-month postintervention, marginally significant at 6-months postintervention, and highly significant at 12-months postintervention. The effect of the intervention for motivation toward alcohol use was significant at 1 month postintervention and marginally significant at 6-months postintervention; however, the intervention had no effect on smoking motivation at any of the three time intervals. The effect of the intervention for improving smoking behavior was marginally significant at 6-months postintervention and was marginally significant at 12-months postintervention for alcohol use. CONCLUSIONS: The intervention had a dampening effect on the upward trajectory of substance use, a path that is well-known to increase with age for both genders in the general population. This short, five-hour program for improving decision making and affecting substance use of teen survivors shows promise; however, a larger sample is needed to enhance findings. IMPLICATIONS FOR NURSING PRACTICE: Besides tailoring risk-behavior information based on actual or potential late effects of treatment to each teen survivor during follow-up visits, oncology professionals need to provide booster programs to refine decision-making skills within meaningful decision context for teen survivors as a means of reducing risk behaviors. PMID- 11064880 TI - An evaluation of the permeability of chemotherapy gloves to three cancer chemotherapy drugs. AB - PURPOSE/OBJECTIVES: To evaluate the permeability of chemotherapy gloves when using carmustine (BCNU), etoposide, and paclitaxel, which were selected based on their reported toxicity and unique solvent systems. SAMPLE: Thirteen brands of chemotherapy gloves and one brand of examination glove. Of the 14 glove types tested, 11 were made of latex, and 3 were made of nitrile. METHODS: Ten samples of each type of glove were evaluated using rigorous laboratory test conditions usually not encountered in normal usage. The thickness of the gloves was measured using a digital caliper. The glove material was secured over glass vials containing the drug solution and inverted in plastic cell wells containing a filter paper disc. After a two-hour exposure time, the filter paper discs were removed and analyzed for the presence of the drug. MAIN RESEARCH VARIABLES: Permeability (i.e., greater than or equal to 1% of the total amount of drug passing through the glove material.) FINDINGS: All 14 types of gloves tested were impermeable to BCNU at two hours of exposure. Only two gloves, the Ansell Perry EP glove and the U.S. Clinical Chemo Bloc T glove, were impermeable to all three drugs. The remaining 12 gloves all demonstrated some level of permeation with etoposide at two hours, although 9 of the gloves had only 1 of 10 samples that were permeable. In all cases, percent of permeation was less than 2% of the amount of the drug in the test solution. Thirteen gloves tested for paclitaxel permeability were impermeable at the two-hour time period. CONCLUSIONS: The results of this study indicate that most of the chemotherapy gloves on the market are either impermeable or minimally permeable to these three chemotherapy drugs. IMPLICATIONS FOR NURSING PRACTICE: Because gloves are universally recognized as a means of personal protection when handling cancer chemotherapy drugs, selection of a glove that is impermeable would be an obvious choice for healthcare workers. Although the present study was a static, laboratory-based study that did not duplicate actual work practice conditions. It should offer some guidance in the selection of glove types when handling chemotherapy drugs. PMID- 11064881 TI - Improving pain outcomes of hospice patients with cancer. AB - PURPOSE/OBJECTIVES: To evaluate the success of a multifaceted, hospicewide, nurse focused pain-management intervention for improving patient pain outcomes. DESIGN: Comparative, descriptive. SETTING: A large, nonprofit hospice that primarily provides home care. SAMPLE: Two samples were included in the study. One sample (n = 47) was from a study completed in 1995 before the intervention, and one (n = 255) was from a study completed in 1997 after the intervention. All patients had been diagnosed with cancer and were alert and able to self-report. METHODS: Secondary analysis of data that were collected as part of two quality-of-life studies. The four-part intervention included intensive pain-management education for the nurses, development and implementation of pain-management policies and procedures, changes in pain assessment and management documentation, and development and use of quality assurance monitors by the nursing staff. MAIN RESEARCH VARIABLES: Pain at its worst, pain relief, and quality of life. FINDINGS: Adjusted mean pain-relief scores were significantly lower in 1995 (X = 5.8) than in 1997 (X = 8.4). In 1995, 43% of patients reported pain relief at a level of 5 or less (on a 0-10 scale). This number dropped to 10% by 1997. Adjusted mean pain-at-its-worst scores were significantly lower in 1997 (X = 6.1) than in 1995 (X = 6.7). Pain relief was found to be positively correlated (r = 0.41-0.51) with quality of life in both samples. CONCLUSIONS: The hospicewide pain-management intervention was effective. IMPLICATIONS FOR NURSING PRACTICE: Through careful study and multifaceted nurse-focused interventions, pain outcomes of hospice patients with cancer can be improved. PMID- 11064882 TI - Coping strategies and psychological distress in patients with advanced cancer. AB - PURPOSE/OBJECTIVES: To examine the relationship between coping mechanisms and psychological distress in patients with advanced cancer receiving chemotherapy. DESIGN: Descriptive, correlational study. SETTING: Private and public hospitals in New York, NY. SAMPLE: 132 patients, ages 33-83, with advanced breast, ovarian, lung, colorectal, or other cancers. METHODS: Mail survey using the Ways of Coping Inventory-Cancer Version and the Profile of Mood States. MAIN RESEARCH VARIABLES: Coping strategies, psychological distress. FINDINGS: The coping strategies distancing, cognitive escape-avoidance, and behavioral escape-avoidance were related to psychological distress. Distancing was negatively related (r = -0.25) and cognitive escape-avoidance (r = 0.38) and behavioral escape-avoidance (r = 0.38) were positively related to psychological distress (p < 0.01). Collectively, the coping strategies explained 36% of the variance of psychological distress. The most important coping mechanism contributing to overall psychological distress was behavioral escape-avoidance, followed by cognitive escape-avoidance. CONCLUSIONS: Low levels of psychological distress were associated with low levels of cognitive and behavioral escape-avoidance and high levels of distancing. IMPLICATIONS FOR NURSING PRACTICE: Nursing interventions that reduce cognitive and behavioral escape-avoidance and enhance distancing should be tested, through further research, in relation to their ability to decrease psychological distress in patients with advanced cancer. PMID- 11064883 TI - The evolving meaning of cancer for long-term survivors of breast cancer. AB - PURPOSE/OBJECTIVES: To discover the different meanings of cancer for older women who are long-term survivors of breast cancer. DESIGN: Qualitative study using a heuristic approach. SETTING: Large metropolitan area in the Midwestern United States. SAMPLE: A sample of eight women was obtained using network sampling. The women ranged in age from 65-77 years. Length of survival ranged from 5.5-29 years. Five of the women had been treated with a lumpectomy (four with radiation and chemotherapy and one with radiation only). The three other women had been treated with a simple mastectomy, one of whom also was treated with chemotherapy. METHODS: Interviews were conducted in the women's homes. Audiotaped interviews were transcribed and manually coded for patterns and themes. MAIN RESEARCH VARIABLES: Meaning of cancer. FINDINGS: Three meanings of cancer emerged from the data: (a) cancer as sickness and death, (b) cancer as an obstacle, and (c) cancer as transforming. CONCLUSIONS: As the women worked through their cancer experience, their perspectives changed. The meaning of cancer after surviving the disease and its treatment centered around positive, insightful experiences and expansive, renewing interactions with their environment. Further research examining the meaning of cancer is needed to broaden the transferability of the findings to other groups. IMPLICATIONS FOR NURSING PRACTICE: Understanding the meaning of cancer for older women who are long-term breast cancer survivors may enhance nurses' sensitivity to survivors' perspectives. Knowledge of survivors' different meanings of cancer may help to paint a new vision of cancer survivorship comprised of potentially positive, transforming experiences. PMID- 11064884 TI - [Experimental research on the usefulness of the Dallos polyester prostheses in reconstructive operations of knee joint ligaments]. AB - Usage of allogenic grafts in surgery of severe injuries of knee joint may be necessary in spite of their disadvantages. Prosthesis Dallos made by Tricomed in Lodz has been tested taking into account its usage in reconstruction of knee sacral ligaments. Prosthesis was implanted into 7 piglets in place of the removed front sacral ligament. In the tests the mechanical requirements made for this kind of grafts were confirmed. However, an unfavourable influence of this prosthesis on the synovial membrane of a joint and weak overgrowth of the graft with connective tissue, which can be eliminated by usage of looser knitting, were noticed. PMID- 11064885 TI - [Testing the usefulness of a provisional polyester prosthesis for augmentation in reconstructive operations of knee joint ligaments]. AB - Prosthesis from polyester fibres 5 mm wide with loose knitting finished on both ends with teflon handles for fixing bones with a wholly aphid screw made by Tricomed company in Lodz according to its own idea was the subject of the test. The tests were made on 7 piglets. The prosthesis together with an autogenic graft taken from the middle third of a patella ligaments was implanted into the place of the excised front sacral ligament. The implanted prosthesis was characterised with a high biocompatibility, satisfying mechanical resistance and easiness of fixing of the tested graft. We noticed clinically and pertaining to autopsy the possibility of removing the allogenic graft after achieving the full healing of the autogenic graft without the possibility of hurting that last one. PMID- 11064886 TI - Gravitational effects in the passive osmotic flows across polymeric membrane of electrolytic solutions. AB - Results of experimental study of volume flux in one-membrane system were presented. This system contains horizontal, microporous and symmetrical flat polymeric membrane (Nephrophan), which separate water and electrolyte solution. As binary solutions, aqueous ammonia solutions, which density is lower than water density, were used. As ternary solutions the ammonia with KC1 (0.1 or 0.2 mole.l 1) in aqueous solution were used. The density of ternary solutions was lower, higher or the same as water density. Two configurations of membrane system (A and B) in gravitational field were studied. In configuration A, water was in compartment over the membrane and the solution was under the membrane. In configuration B the succession was reverse. The thermodynamic model of flux graviosmotic effect was elaborated, and the calculations of this effect were performed for A and B configurations of one membrane system. The experimental results are interpreted in terms of gravitational instability that reduces concentration boundary layer dimensions and increases the diffusion permeability coefficient value of the complex: boundary layer/membrane/boundary layer. PMID- 11064887 TI - [Biological tests of sol-gel biomaterials]. AB - Recently, the sol-gel based biomaterials are extendedly investigated in emphasis on theirs medical applications. In this respect it is important to investigate the influence of sol-gel matrices on biological systems. The results of laboratory and biological testing of water extracts of sol-gels are presented in this work. It was proved that it is possible to construct the sol-gels that are not cytotoxic for which the haemolytic reactions fulfils the foreseen norms. This can be achieved by heating the materials in certain temperatures (higher than 350 degrees C). This effect can also be reached by suitably long aging (minimum 6 months). PMID- 11064888 TI - [Nutrition and prostatic cancer]. AB - Due to the major geographical variations affecting its clinical incidence, prostate cancer appears to be influenced by environmental factors, which may either promote or inhibit the development of this tumour. Diet appears to play a considerable role among these environmental factors. There are epidemiological and experimental arguments in favour of the role of diet in the development of prostatic cancer. Certain foods, such as fats, phenolic compound and other micronutrients such as vitamins or selenium have been reported to have an action on the natural history of prostate cancer. The authors present a review of the literature analysing the various potential actions of various foods. PMID- 11064889 TI - [Simultaneous radio-chemotherapy for infiltrating bladder carcinoma. What is new since 1997?]. AB - Concomitant radiotherapy-chemotherapy is increasingly used in urology in the treatment of invasive bladder tumours. In 1997, the outcome of 552 patients was reported in the international literature. Two years later, another 617 patients had been treated according to various modalities. The initial complete response rate ranged from 56 to 87% (median: 70%). 5-year overall survival rates were between 55 and 68% for T2 tumours. The metastasis rate was between 22 and 35% for the overall treated population. Concomitant radiotherapy-chemotherapy using a platinum salt represents a possible therapeutic modality in some patients, but it cannot be considered to be equivalent to total cystectomy, which remains the reference treatment in operable patients with an operable tumour, achieving a 63 to 80% 5-year survival for pT2 tumours. PMID- 11064890 TI - [Remote laparoscopic radical prostatectomy carried out with a robot. Report of a case]. AB - Robotics has been commonly employed in numerous industrial fields for several decades. However, the application of this technology to surgery is a recent innovation. It provides new possibilities for facilitating specific surgical tasks, especially in the field of laparoscopy. We report a case of laparoscopic radical prostatectomy completed with the help of a remotely controlled da Vinci robot. This system offered a user friendly surgical platform and enhanced surgical dexterity. Operating time was 420 minutes and the hospital stay was 4 days. The bladder catheter was removed after 3 days. One week later, the patient was fully continent. Pathologic examination showed a pT3a tumor, with negative margins. Robotically assisted laparoscopic radical prostatectomy is feasible. Further developments in this field of technology may have new applications in laparoscopic telesurgery. PMID- 11064891 TI - [Contribution of lumboscopy in the treatment of pyeloureteral junction syndromes. Report of 25 cases]. AB - OBJECTIVE: Evaluation of the results of lumboscopic repair of ureteropelvic junction syndromes. MATERIAL AND METHOD: Retrospective study of 25 consecutive lumboscopic retroperitoneal pyeloplasties performed over 3 years in 14 women and 11 men with symptomatic ureteropelvic junction syndrome. RESULTS: The mean operating time was 200 minutes (range: 120-360 minutes) and mean blood loss was 60 ml. Surgical conversion was necessary in three cases due to the difficulty of dissection and in one case because of rupture of the ureter. Analgesic prescription was generally only necessary for the first 2 postoperative days. The mean hospital stay was six days (range: 2-16 days). Patients were able to return to work an average of 10 days after the operation. With a mean follow-up of 9 months (range: 6 to 18 months), all patients were asymptomatic except for one patient who reported pain at a trocar site. Follow-up urography at 3 months showed marked improvement in 18 patients (85.7%), moderate pyelocaliceal dilatation in 2 cases and one failure. CONCLUSION: Lumboscopic pyeloplasty is an alternative to endopyelotomy. It provides a comparable success rate to that of conventional surgery, while reducing the morbidity, length of hospital stay and convalescence. However, it requires mastery of intracorporeal suture techniques. PMID- 11064892 TI - [Predictive factors of successful treatment of lower caliceal calculi with Edap LT02 extracorporeal lithotripsy]. AB - OBJECTIVE: To determine the predictive factors of success of extracorporeal shock wave lithotripsy (ESWL) for lower caliceal stones. PATIENTS AND METHODS: The case files of 100 patients with a single stone in the lower pole of the kidney, ranging in size from 6 to 25 mm, treated by EDAP LTO2 extracorporeal lithotripsy between 1994 and 1997, were studied. Pretreatment intravenous urography was reviewed to assess the characteristics of the stone, to measure the pyelocaliceal angle and to study the anatomy of the lower pole of the kidney. RESULTS: The overall stone-free rate at 3 months was 57%. The success rate was 67.18% for stones smaller than 1 cm and 38.88% for stones larger than 1 cm. Stones denser than bone were successfully treated in 45.5% of cases, those less dense than bone were successfully treated in 71.11% of cases. The success rate was 86.04% when the pyelocaliceal angle was greater than 90 degrees and 35.08% when this angle was less than 90 degrees. The stone-free rate was 75% when the caliceal stalk was less than 3 cm and 37.5% when the stalk was greater than 3 cm. CONCLUSION: In this series, the size of the stone, its density, the pyelocaliceal angle and the length of the caliceal stalk were predictive elements of the success of ESWL for lower caliceal stones. The pyelocaliceal angle was the most significant factor. A very dense stone on the plain abdominal film with a diameter of 1 cm or more and presenting unfavourable anatomical factors should be treated by percutaneous nephrolithotomy as the first-line procedure. PMID- 11064893 TI - [Does ureteroscopy still play a role in the treatment of ureteral calculi?]. AB - OBJECTIVES: To evaluate the results obtained by ureteroscopy to treat ureteric stones and the related complications. MATERIALS AND METHODS: 74 ureteroscopies were studied retrospectively, including 62 cases of pelvic ureteric stones. In situ fragmentation was necessary in 28.4% of cases. Success was defined as the absence of stones for one month after ureteroscopy. Success rates were analysed according to size and site of the stone, the need for fragmentation and the patient's sex. RESULTS: Morbidity was 6.8%. Success was obtained in 60.8% of cases: 72.7% in women and 55.8% in men. The mean diameter of successfully treated stones was statistically smaller (6.2 +/- 2.1 mm) than that of stones with a poor result (7.8 +/- 3.7 mm) (p = 0.044, Spearman). CONCLUSIONS: The size and site of the stone and the patient's sex influence the results of ureteroscopy. Ureteroscopy for stone disease remains a topical modality after failure of extracorporeal lithotripsy, but also in the case of poor access of an effective lithotriptor. PMID- 11064894 TI - [Intravesical BCG-therapy: comparison of side effects of Connaught (Toronto) and Pasteur (Paris) strains]. AB - INTRODUCTION: Urologists have felt that the adverse effects of intravesical BCG therapy have been more serious and more frequent since the use of the Connaught strain. The objective of this retrospective study was to compare the toxicity of this new strain with that previously used in France (Pasteur strain). MATERIAL AND METHODS: After endoscopic resection, 89 patients with stage Ta grade 1-2 recurrent or T1 grade 3 and/or CIS bladder tumour were treated with 6 instillations of 150 mg of BCG Pasteur from 1992 to 1996 (50 patients: group 1) or 81 mg of BCG Connaught from January 1997 to December 1998 (39 patients: group 2). Adverse effects were classified as minor, lasting less than 48 hours (bladder irritation syndrome and/or macroscopic haematuria and/or fever less than 38 degrees C), moderate (requiring symptomatic treatment, reduction of the dose or an increased interval between instillations), and major (contraindication to continuation of treatment). RESULTS: 74% of patients in group 1 presented at least one adverse effect versus 77% in group 2. The reasons for permanent discontinuation of BCG-therapy in groups 1 and 2, respectively, were as follows: malaise during instillation (1 vs 0), bladder irritation syndrome not controlled by symptomatic treatment (4 vs 5) and epididymitis (0 vs 1). Pulmonary tuberculosis was diagnosed in one patient from group 2, one year after the last instillation. The frequency and severity of adverse effects were not statistically different between the two groups. The number of patients discontinuing BCG-therapy because of severe complications was also not statistically different between the two groups. CONCLUSION: This study did not reveal any difference of toxicity between Connaught and Pasteur strain in intravesical BCG-therapy of superficial bladder tumours. PMID- 11064895 TI - [Assessment of urinary continence in Hautmann neobladder]. AB - OBJECTIVE: The Hautmann neobladder is a bladder replacement technique frequently proposed after total cystectomy for bladder cancer. The objective of this prospective study was to evaluate the patients' urinary continence after this operation, based on clinical and urodynamic data. MATERIALS AND METHODS: The functional assessment was based on 26 patients from a series of 45 consecutive Hautmann bladder replacements performed between February 1994 and May 1999. These 26 nonselected patients (21 men and 5 women) with a mean age of 56 years (range: 38-68) accepted the principle of functional assessment of continence at visits held 1, 3, 6 and 12 months postoperatively including clinical interview, urodynamic studies and fibroscopy. Only one patient presented with preoperative stress incontinence. The mean follow-up of these 26 patients was 22 months (range: 10-60). Three patients, including two women, died from progression of their bladder tumour at the 12th, 20th and 32nd postoperative months. RESULTS: Urodynamic studies showed a mean maximum capacity of the ileal reservoir of 420 ml (range: 316-571), a maximum filling pressure of 15 cmH2O (range: 2-24) and a maximum urethral closure pressure of 49 cmH2O (range: 31-74). According to the evaluation criteria used, the satisfactory continence rates, as assessed by the patients were 62%, 77%, 84.6% during the day and 42%, 615%, 77% at night, at 3, 6 and 12 months, respectively. CONCLUSION: The Hautmann neobladder ensures satisfactory diurnal and nocturnal urinary continence in more than 75% of cases after the 6th postoperative month. Continence is an evolving parameter especially during the first postoperative year. The selection of patients in good general condition and motivated for management of their new urinary situation remains an essential prerequisite to obtain a good functional result. PMID- 11064896 TI - [Study of survival after cystectomy for bladder cancer. Report of 504 cases]. AB - OBJECTIVES: Cystectomy is the reference treatment for invasive bladder cancer and superficial tumours with a high risk of recurrence. However, the long-term results of this treatment remain controversial. Progress in anaesthesia-intensive care and surgical techniques appear to have improved the prognosis of this disease over the last two decades. The availability of numerous adjuvant therapies (radiotherapy and chemotherapy) and the development of alternative conservative management therefore require a re-evaluation of the long-term results of cystectomy for bladder cancer performed over the last 20 years. MATERIAL AND METHODS: The case files of 504 consecutive patients undergoing cystectomy for bladder cancer in our department from 1981 to 1997 were reviewed. The operative and postoperative morbidity and actuarial survival by stage were studied. Histological prognostic factors and the influence of adjuvant therapies were also studied. RESULTS: According to the TNM 97 classification, 55% of tumours (on the cystectomy specimen) were intravesical (< T3), and 70% of patients had negative lymph nodes (N0). The perioperative mortality was 1.56%. The overall survival at 2 years, 5 years and 10 years for the total patient population was 83.1%, 52.3% and 46.6%, respectively. The 5-year survival of tumours confined to the bladder (< T3) was 79.4% versus 27.5% when the tumour extended beyond the bladder (> T3). The lymph node status considerably influenced survival. N0, N1 and N2-3 patients had 5-year survival rates of 64%, 48% and 14%, respectively. Neoadjuvant chemotherapy or radiotherapy did not appear to improve survival. CONCLUSIONS: Survival after cystectomy for bladder cancer essentially depends on pathological stage and lymph node status. Patients with a localized tumour have a 5-year survival greater than 80%. Prospective studies are required to determine the real benefit of adjuvant chemotherapy, as its value has not yet been formally demonstrated. PMID- 11064897 TI - [Symptom scores: what do they describe exactly?]. AB - OBJECTIVE: Standardized symptom scores have been developed to evaluate voiding disorders. The most widely used in France are the IPSS and Madsen-Iversen. These self-assessment questionnaires are by definition subjective. The objective of this study was to compare the capacities of the IPSS and Madsen-Iversen score to describe the patient's voiding status and to define the patient's level of understanding of these questionnaires. PATIENTS AND METHODS: One hundred male patients were included in this prospective study. All completed an IPSS and Madsen-Iversen questionnaire. Physical examination, uroflowmetry and post-voiding bladder ultrasonography were also performed. RESULTS: The description of the voiding status was considered to be satisfactory or fairly satisfactory, with a total of 85% for IPSS and 87% for Madsen-Iversen. Understanding of the questionnaire was high with a value of 84% for IPSS and 83% for Madsen-Iversen. Neither the order of completion of the IPSS or Madsen-Iversen questionnaires nor the patient's age influenced these results. CONCLUSIONS: No difference was demonstrated between the IPSS score and the MADSEN-IVERSEN questionnaire in terms of description and comprehension. However, one out of five patients experienced difficulties completing these questionnaires. PMID- 11064898 TI - [Prevalence of benign hypertrophy of the prostate in Turkish men hospitalized in urology]. AB - OBJECTIVES: The prevalence of clinical benign prostatic hyperplasia (BPH) is known in Western European countries and in North America, but epidemiological data are lacking for Eastern European and Balkan countries. The objective of this study was to determine the prevalence of BPH in this region. METHODS: We analysed the data obtained on patients with BPH reported by urologists from 48 hospitals in Turkey, in 1996. This analysis is based on 84,149 case files for patients aged 40 to 80 years and more. RESULTS: The overall prevalence of clinical BPH was 18.5%. The age-specific predominance of BPH reached a maximum for the 60-69 years age-group. A majority of cases of BPH were treated medically (54.7% of cases). CONCLUSION: The data are discussed in the light of our current knowledge. Our presentation is the first report concerning the prevalence of clinical BPH in our region, but it needs to be completed to determine the prevalence of clinical BPH in relation to age, country and race in the Balkans and the Middle East. PMID- 11064899 TI - [Is it possible to define a threshold for free PSA index that is useful in the daily practice of urology?]. AB - OBJECTIVES: Retrospective evaluation of the use of the free PSA index before prostatic biopsies. MATERIAL AND METHODS: The authors retrospectively studied the values for total PSA, free PSA, and free PSA index (ratio of free PSA over total PSA expressed as a %) in men with a total PSA between 2 and 10 ng/ml, from a population of 391 men prior to prostatic biopsies. They also isolated a subgroup of patients in whom the free PSA index could have been used as a first-line marker to decide whether or not to perform prostatic biopsies. RESULTS: The mean values for total PSA, free PSA, and free PSA index were compared as a function of the diagnosis, age, and ultrasound prostatic volume. The yields of the various cut-off values for the free PSA index for PSA between 2 and 4 ng/ml, 4 and 10 ng/ml, and 2 and 10 ng/ml with a normal digital rectal examination are reported. Between 2 to 10 ng/ml, at a cut-off value of 30%, 94.1% of cancers would have been detected (sensitivity) and 22% of biopsies would have been avoided, 10 of which would have been useless, i.e. a 30.3% economy of useless biopsies not performed (specificity). At the cut-off value of 15%, less than half of cancers would have been detected (47.1%) and 90.9% of useless biopsies would have been avoided. Biases creating difficulties of interpretation were the assay kits, the reference population, age, storage of sera, and prostatic volume. CONCLUSION: The free PSA index would be a useful first-line parameter in only 12.7% of candidates for prostatic biopsies. The cut-off value of 30%, validated for our assay method, would be able to detect the majority of cancers in men aged 50 to 65 years, while avoiding biopsies in the third of men with no detectable cancer. PMID- 11064900 TI - [Retroperitoneal lymphadenectomy for testicular cancer and genito-sexual conditions: retrospective study]. AB - OBJECTIVES: The aim of this study was to evaluate retrospectively the preservation of fertility in a number of patients with testis or funicle tumour treated with retroperitoneal lymph node dissection (RPLND). MATERIAL AND METHODS: Between 1983 and 1998, 41 patients with testis or funicle cancer (mean age 29 years, range 18-58) underwent RPLND at our institution. Clinical staging included abdominal CT scan, chest X-rays and serum tumour markers (alpha FP, beta HCG, LDH). RPLND was performed bilaterally in 14 patients and unilaterally in 13 patients (6 right and 7 left). The nerve sparing technique was used in 14 cases. Ejaculation was evaluated in 39 patients (2 patients died of metastases before the study). Mean follow up was 64 months (range 5-182). Semen was available for 21 patients before RPLND and for 19 patients after RPLND. The "t Student" test was used to compare the semen parameters before and after surgery. RESULTS: Bilateral RPLND caused loss of ejaculation in 67% of the patients (8/12). Unilateral right and left RPLND allowed to maintain ejaculation in 100% (6/6) and 57% (4/7) of cases respectively. Nerve sparing procedure preserved ejaculation in 100% of the patients (14/14). After RPLND, both mean total sperm count and mean motility rate were not significantly changed (143 +/- 124 x 10(6) vs 128.2 +/- 72 x 10(6) p > 0.05; 40.7 +/- 17.6 vs 48 +/- 15.5%, p > 0.05). The survival rate of the patients treated with RPLND was 95%. None of the patients treated with unilateral (en bloc or nerve sparing) RPLND had relapse. CONCLUSION: The evolution of surgical technique has notably reduced the andrological complications of the RPLND without affecting the oncological results. PMID- 11064901 TI - [Adrenal gland ganglioneuroma]. AB - The authors report a case of adrenal ganglioneuroma in a 25 year old woman. This rare and benign tumor is localized in the adrenal gland in 20% of cases. Considerations on diagnosis, pathogenesis and management are discussed. PMID- 11064902 TI - [Renal actinomycosis with fistulized lumbar abscess]. AB - The authors report a case of renal actinomycosis in an adolescent presenting with two fistulized lumbar abscesses. This rare disease, which generally has a good prognosis, is difficult to diagnose, both clinically and radiologically. The positive diagnosis is based on histological examination, more frequently of the nephrectomy operative specimen, than ultrasound-guided fine needle aspiration biopsy of an atypical renal tumour. Conservative treatment with high-dose penicillin gives excellent results. PMID- 11064903 TI - [Primary neuroendocrine carcinoma of the bladder: diagnosis and treatment]. AB - OBJECTIVE: This rare but aggressive bladder tumour presents the morphological and immunohistochemical characteristic, common to all neuroendocrine tumours observed in other organs. This study analyzed the diagnostic criteria and therapeutic results obtained in 5 consecutive patients over a 3-year period. MATERIALS AND METHODS: 5 patients (3 men and 2 women) suffering from primary small cell carcinoma of the bladder were evaluated. Histological diagnosis, treatment modalities and outcome were reviewed. RESULTS: The main clinical presentation was macroscopic haematuria. All tumours were invasive at the time of diagnosis. 4 patients were treated by trans urethral resection alone, 2 of whom also received adjuvant radio-chemotherapy. One patient was treated by radical cystectomy. The 4 patients treated by conservative treatment modality had progression and a shorter survival, in contrast with patient treated by radical cystectomy. CONCLUSION: The urologist must recognize this rare histological entity, which have a poor prognosis and requires multidisciplinary management. Treatment must consist of a combination of neoadjuvant or adjuvant chemotherapy and surgery to achieve the best results. PMID- 11064904 TI - [Spontaneous intraperitoneal rupture of the bladder treated with laparoscopy]. AB - The authors report the case of a 53-year-old patient admitted for spontaneous bladder rupture occurring during an episode of alcoholic intoxication. Laparoscopic treatment consisted of peritoneal toilet with bladder suture and drainage of the abdominal cavity with a favourable postoperative course. PMID- 11064905 TI - [Prostatic malacoplakia: report of 3 cases]. AB - Malakoplakia is a granulomatous inflammatory disorder occurring rarely in the prostate. We report our experience with three patients aged from 54 to 75 years old. Clinical presentations were unspecific. All patients presented with a past history of urinary tract infection. Ultrasound study revealed hypoechoic peripheral zone lesions. Histological examination showed a diffuse granulomatous inflammation with numerous histiocytes containing Michaelis-Gutmann bodies. In the prostate, malakoplakia is a histological variant of granulomatous chronic prostatitis following urinary infections. Clinically and radiologically, the differential diagnosis with adenocarcinoma is difficult. The symptoms disappear with a prolongated antibiotic treatment. PMID- 11064906 TI - [Epidermoid carcinoma of the male urethra]. AB - Squamous cell carcinoma of the male urethra is exceptional, as all urethral tumours represent less than 1% of urinary tract tumours. Treatment depends on the stage and site of the lesion, but the prognosis remains very poor despite aggressive treatment, including mutilating resection surgery with or without associated radiotherapy. However, the current hope for patients with squamous cell carcinoma of the urethra resides in radiotherapy-chemotherapy combination protocols based on the results obtained in squamous cell cancers of the oesophagus and anus. PMID- 11064907 TI - [Epidermoid cyst of the testis. Conservative surgery in 3 cases]. AB - The authors report 3 cases of epidermoid cyst of the testis. Clinical, laboratory and ultrasound data were relatively nonspecific. Precise histological criteria were defined by PRICE in 1969. The diagnosis is based on frozen section examination after transnnguinal testicular exploration. The authors describe the most conservative therapeutic approach for a constantly benign testicular tumour. PMID- 11064908 TI - [Statistics and deception. Methodology to help the reader]. PMID- 11064909 TI - [The causal relationship]. AB - Only the controlled trial method, clinical equivalent to the experimental method, with its successive phases and randomization, is able to confirm a real causal relationship and quantify the risk of error (alpha). However, the study must have sufficient power and randomization must not have resulted in an unbalanced distribution of various parameters likely to influence the result. Other methods, particularly surveys and case studies, only provide presumptions of causality. This review article, illustrated by three examples from the urological literature, is designed to demonstrate the difficulties of establishing a causal relationship when possible biases and confounding factors are taken into account. PMID- 11064910 TI - [Surgical treatment of urethral stricture. Personal view]. AB - The author expressed his personal views on the indications, advantages and disadvantages of current techniques for the treatment of urethral stricture in men, such as: endoscopic urethrotomy, end-to-end anastomosis, prepuce or oral mucosa free grafts, penile or scrotal pedicled flaps, expansible metal stents and two-stage urethroplasties. Apart from his own personal experience, he bases his discussion on the pathophysiology of healing and, often old data of the literature concerning surgery of the urethra. He critically reviews recent developments: the posterior approach to urethral stricture and temporary stents. Finally, he presents two personal techniques: endoscopic urethroplasty with free graft around a biodegradable stent and the use of a pudendal fasciocutaneous flap to correct large perineal defects. PMID- 11064911 TI - [Free PSA: its routine use is premature in the screening of prostatic cancer]. AB - The free form of PSA represents an average of 30% of total PSA. The free PSA level, in relation to total PSA, is expressed as a percentage (of free PSA). The percentage of free PSA appears to be relatively independent of benign prostatic hypertrophy, but it is markedly and significantly decreased in the case of prostate cancer and acute prostatitis. A cut-off value for the percentage of free PSA combined with a cut-off value for total PSA can decrease the number of biopsies indicated for the detection of prostate cancer. This approach avoids 20 to 30% of useless biopsies (specificity), but is accompanied by a 5 to 10% reduction of the number of cancers detected (sensitivity of 90 to 95%) compared to the use of total PSA alone. This 5 to 10% risk of missing a cancer of significant volume in a man under the age of 65 years explains why the use of free PSA to guide prostatic biopsies is not routinely recommended. Despite the promising performance of the percentage of free PSA to improve the indication for prostatic biopsies, the methodology of the studies performed to evaluate this test is not sufficient to validate their conclusions. The percentage of free PSA can be prescribed as a second-line test by the urologist, following a first series of negative biopsies in a man with a high clinical and laboratory suspicion of prostatic cancer, in order to propose a second series of biopsies after three months or to define the frequency of clinical and laboratory surveillance. While waiting for the results of prospective studies in the screening setting, recommendations concerning the use of PSA by general practitioners, who are the first to order this test in the context of screening, can be formulated as follows: The value of free PSA assay for first-line cancer screening has not been validated. A consensus has not been reached concerning the use of free PSA as an indication for biopsies and the frequency of assays. Total PSA assay (cut-off value: 4 ng/ml) remains the reference laboratory test for screening and indication of biopsies. A lower normal cut-off value (between 2 and 4 ng/ml) is currently under evaluation. PMID- 11064912 TI - [Repair of female urinary incontinence with prolene "TVT": preliminary results of a multicenter and prospective survey]. AB - OBJECTIVES: A multicentre, prospective study (6 private centres, 1 general hospital and 1 teaching hospital) was conducted to evaluate the perioperative morbidity and short-term functional results of the TVT procedure in the treatment of the female urinary stress incontinence. PATIENTS AND METHODS: From November 1996 to September 1999, 120 patients with a mean age of 65.2 years (range: 37-91) were operated according to the tension-free vaginal tape (TVT) technique for isolated urinary stress incontinence (stage 2 or 3) in 94 cases and associated with pelvic tone disorder in 26 cases. 59 patients (49.2%) presented recurrence of urinary incontinence that had already been operated between 1 and 4 times. Physical examination demonstrated hypermobility of the urethra in 73 cases (60.8%), isolated clinical sphincter incompetence in 47 cases (39.2%) and pelvic tone disorders in 31 cases. Urodynamic studies, performed in 113 patients, demonstrated sphincter incompetence in 65 cases (57.5%) with a mean maximum urethral closure pressure of 18 cmH2O (range: 5-29). RESULTS: The operation, performed under spinal anaesthesia in 97 cases (80.8%), general anaesthesia in 16 cases (13.3%) and local anaesthesia in 7 cases (5.8%) lasted an average of 28.7 min (range: 15-60) for insertion of the TVT. Perioperative complications consisted of twelve bladder injuries (10%) and two pelvic haematomas (1.7%). No cases of infection, erosion or migration of the tape were reported. In the group of 94 patients operated exclusively by TVT, the mean hospital stay was 2.6 days (range: 1-7). Twelve patients (10%) required self-catheterization for 2 to 30 days. With a mean follow-up of 15.2 months (range: 36-6), continence was restored in 104 patients, corresponding to a cure rate of 86.7%. A marked improvement was obtained in 11 cases (9.2%) and five cases (4.2%) were considered to be failures. CONCLUSION: The TVT procedure is a new approach to the treatment of female urinary stress incontinence. Its advantages are its simplicity, the rapidity of the technical procedure and the short-term efficacy on continence. A longer follow-up is essential to assess to the functional outcome and the long-term urethral tolerance. PMID- 11064913 TI - [Treatment of chronic urinary retention after surgical treatment of urinary incontinence with bladder neck transurethral resection]. AB - OBJECTIVE: To evaluate the efficacy and safety of transurethral resection of the overcorrected posterior lip of the bladder neck in patients with chronic urinary retention after repair of incontinence. MATERIAL AND METHODS: Transurethral bladder neck resection was performed in 26 women with a median age of 59 years. Incontinence repair consisted of a Burch procedure in eight cases, a Raz procedure in eight cases, a Marshall-Marchetti-Krantz procedure in five cases, an aponeurotic sling in three cases and a synthetic sling in two cases. The median preoperative maximum urine flow rate was 11.5 ml/s and the median residual urine was 150 ml. Preoperative cystourethrography and cystoscopy revealed overcorrection of the bladder neck in each case. RESULTS: With a median follow-up of 39 months, 65.5% of patients were cured (resolution of symptoms, maximum urine flow rate greater than 15 ml/s and residual urine less than 50 ml), 23% were improved and 11.5% were considered to be failures. No complications or secondary urinary incontinence were observed. Patients not cured by this technique were treated by urethrolysis in three cases, Uroflow stent in one case and section of a Raz cervicocystopexy suture in one case. CONCLUSION: Transurethral bladder neck resection can be used as first-line treatment for chronic urinary retention after repair of incontinence, as it is an effective, rapid, minimally invasive technique not associated with any morbidity. Urethrolysis can always be performed in the case of failure. PMID- 11064914 TI - [Vesico-uterine fistulae. Report of 5 cases]. AB - The authors report 5 cases of uterovesical fistula (UVF) and analyse some of the clinical and therapeutic aspects of this disease. The mean age of these 5 patients was 31.2 +/- 6.9 years. The trauma responsible for the fistula was caesarean section in every case. The presenting complaints were dominated by cyclic haematuria (n = 5). Intravenous urography did not contribute to the diagnosis, while hysterosalpingography demonstrated the UVF in 3 out of 4 cases. Treatment was surgical for all patients and consisted of closure of the fistula in 3 cases and hysterectomy in 2 cases. With a mean follow-up of 2 +/- 1.2 years, no pregnancies were reported in the 3 patients treated by closure of the fistula. On the other hand, the urinary results were satisfactory with good continence and resolution of the cyclic haematuria. The authors emphasize the importance of prevention of UVF by well conducted caesarean section. PMID- 11064915 TI - [Lower pole pedicle in a series of 84 pyelo-ureteral junction syndromes surgically treated in children]. AB - OBJECTIVE: To evaluate the involvement and consequences of the lower pole pedicle (LPP) associated with ureteropelvic junction syndrome. MATERIAL AND METHOD: Retrospective study in 81 children presenting a total of 84 ureteropelvic junction syndromes operated consecutively between 1994 and 1998. Urinary tract ultrasound and cystography were systematically performed. Renal scintigraphy (DTPA or MAG 3) was performed in 80 children. Preoperative intravenous urography was performed in 60 children. LPP was considered to be present when its participation in the obstruction was confirmed intraoperatively (Anderson Hynes technique with uncrossing of the vascular pedicle). RESULTS: Group I: a LPP was revealed in 24 kidneys (28.5% of cases), 17 left kidneys and 7 right kidneys, in 14 boys and 10 girls, with a mean age of 4 years (range: 2 months-14 years). Group II: 60 kidneys without LPP (71.5% of cases), 32 left kidneys and 28 right kidneys, in 40 boys and 17 girls, with a mean age of 2 years (range: 1 month-15 years). The most frequent presenting complaint was recurrent low back pain in 58% of cases in group I (14/24) and 5% of cases in group II (3/60). The mean age at diagnosis was 6 years. Hydronephrosis was detected by antenatal ultrasound in 33% of cases in group I (8/24) and in 72% of cases in group II (43/60). Kidney function in group I was greater than 40% in 19 patients, between 20 and 39% in 2 patients and less than 20% in 2 patients. These results were not influenced by age at diagnosis and were not significantly different from those observed in group II. Renal malrotation was observed in 2 cases in group I and in 12 cases in group II. Histology of the junction revealed nonspecific fibrosis in the same percentage of cases (91%) in the two groups. The mean follow-up was 15 months (range: 2 months-5 years). No surgical failure was observed. CONCLUSION: Ureteropelvic junction syndrome associated with a LPP appears to present later with recurrent low back pain in older children. It does not worsen the functional prognosis of the affected kidney. LPP can be visualized by duplex ultrasound. It may act as an inducer of obstruction by aggravating a pre-existing abnormality of the ureteropelvic junction. When LPP is associated with isolated dilatation of the pyelocaliceal cavities, the risk of subsequent decompensation requires closer ultrasound surveillance, until puberty. PMID- 11064916 TI - [Treatment of neurogenic urinary incontinence in children]. AB - The current management of neurogenic urinary incontinence is exclusively urological and therefore palliative. However, urological techniques have considerably advanced over the last 20 years and now allow continence with protection of the upper urinary tract in the very great majority of cases. The authors review the various techniques proposed in achieve acceptable continence and define the indications based on published results and their personal experience. PMID- 11064917 TI - [Anterior hypospadias: Duplay or Mathieu?]. AB - OBJECTIVE: The Mathieu urethroplasty is considered to be the reference operation for the treatment of anterior hypospadias. The vertical incision of the urethral groove, described by Snodgrass, has extended the field of indications for urethroplasty by tubularisation of the urethral plate (Duplay principle), by allowing it to be applied in almost every case of anterior hypospadias, The objective of this study was to compare results of Mathieu urethroplasty and Duplay urethroplasty for the treatment of anterior hypospadias. MATERIALS AND METHODS: 50 Mathieu urethroplasties and 50 Duplay urethroplasties were performed by the same operator over the same period (1996 to 1999). All urethral sutures were performed by inverted running sutures. Urine drainage was ensured by Foley catheter for 48 hours. For Duplay operations, the urethral plate was incised longitudinally in 17 out of 50 cases. Foreskin reconstruction was performed 41 out of 100 cases. RESULTS: The follow-up is 6 months to 4 years. Three children (6%) in each group had to be reoperated because of urethral complications (fistula, meatal stricture, urethroplasty dehiscence). CONCLUSION: Our results confirm those published in the literature. Duplay and Mathieu urethroplasties provide equivalent results in terms of the urethra. The appearance of the urinary meatus appears to be more satisfactory after Duplay urethroplasty. The Snodgrass modification allows tubularisation of the urethral plate to be performed even when it is narrow. Cover of the urethroplasty by a foreskin subcutaneous connective tissue pedicled flap reduces the fistula rate of Duplay urethroplasty to the same value as that observed after Mathieu urethroplasty, although it classically used to be higher. PMID- 11064918 TI - [Re: "Testicular cancer and male fertility"]. PMID- 11064919 TI - [Re: "Intraprostatic vascularization in power Doppler imaging and vascular 3D: analysis criteria and role in the diagnosis and staging of cancer"]. PMID- 11064920 TI - Fixation and imaging of biological elements: heavy metals, diffusible substances, ions, peptides, and lipids. AB - We tested various fixation and analysis methods to demonstrate by electron microscopy elemental imaging in tissues and cells, i.e., soluble substances such as many kinds of ionic elements, water soluble low molecular peptides, and even organic solvent soluble substances such as lipids. For the ionic elements, we tested frozen dried or freeze-substituted methods and organic or inorganic special chemical precipitation methods combined with microwaved fixation methods. The data were analyzed with electron beam X-ray microanalysis, electron energy filtered imaging analysis, and electron microscope autoradiography. The data were demonstrated as elemental distribution images and were calculated quantitatively. For the soluble low molecular peptides, we developed a tannic acid and aldehyde method combined with microwaved fixation. We discuss the theoretical background of the tannic acid fixation and microwaved fixation methods. For the organic solvent soluble substances, i.e., lipids including steroids, we successfully tested the use of a mixed fixative of aldehyde and osmium, digitonization, and osmification with the use of p-phenylendiamine or imidazole. We also proposed some new ideal biotracers for electron beam X-ray microanalysis and electron energy filtered imaging analysis. PMID- 11064921 TI - Myotonic dystrophy and myotonic dystrophy protein kinase. AB - Myotonic dystrophy protein kinase (DMPK) was designated as a gene responsible for myotonic dystrophy (DM) on chromosome 19, because the gene product has extensive homology to protein kinase catalytic domains. DM is the most common disease with multisystem disorders among muscular dystrophies. The genetic basis of DM is now known to include mutational expansion of a repetitive trinucleotide sequence (CTG)n in the 3'-untranslated region (UTR) of DMPK. Full-length DMPK was detected and various isoforms of DMPK have been reported in skeletal and cardiac muscles, central nervous tissues, etc. DMPK is localized predominantly in type I muscle fibers, muscle spindles, neuromuscular junctions and myotendinous tissues in skeletal muscle. In cardiac muscle it is localized in intercalated dises and Purkinje fibers. Electron microscopically it is detected in the terminal cisternae of SR in skeletal muscle and the junctional and corbular SR in cardia muscle. In central nervous system, it is located in many neurons, especially in the cytoplasm of cerebellar Purkinje cells, hippocampal interneurons and spinal motoneurons. Electron microscopically it is detected in rough endoplasmic reticulum. The functional role of DMPK is not fully understood, however, it may play an important role in Ca2+ homeostasis and signal transduction system. Diseased amount of DMPK may play an important role in the degeneration of skeletal muscle in adult type DM. However, other molecular pathogenetical mechanisms such as dysfunction of surrounding genes by structural change of the chromosome by long trinucleotide repeats, and the trans-gain of function of CUG binding proteins might be responsible to induce multisystemic disorders of DM such as myotonia, endocrine dysfunction, etc. PMID- 11064922 TI - [Concern with systematic registration of medical data]. PMID- 11064923 TI - Registration of congenital anomalies in Switzerland by EUROCAT. AB - Since 1988 the epidemiological surveillance of congenital anomalies (malformations, chromosomal aberrations, metabolic diseases, hereditary diseases, neurosensorial defects, etc.) is carried out by the Swiss registry of EUROCAT (European Registry of Congenital Anomalies and Twins). Several Swiss cantons collaborate through their own local registry, transmitting data to the central registry in Lausanne. We present the main objectives and methods of registration and give the global prevalence rates for the main malformations for 1996 and the period 1993-1996. PMID- 11064924 TI - [Data on selected prenatal malformations in the EUROCAT study. Results of Zurich Canton from 1988 to 1997]. AB - In the context of the EUROCAT study, data on selected congenital malformations and chromosome aberrations were collected from the Canton of Zurich (1988-1997). It was found that the major proportion of severe and early malformations, such as anencephalus and holoprosencephaly, were detected prenatally; for oral clefts and meningomyeloceles this was not the case, at any rate in regard to isolated (non syndromic) malformations. However, if these defects occur in combination with a chromosome aberration, the likelihood of such a case being registered is higher. For the same reason, i.e. due to abnormal ultrasound findings and intrauterine growth retardation, trisomies 13 and 18 were more often detected prenatally than trisomy 21. PMID- 11064925 TI - [Trisomy 21 and its prenatal detection in the Canton of Vaud (1980-1996)]. AB - We present a genetic and epidemiological study of trisomy 21 (T21) in the Canton of Vaud, the area covered by our local registry of congenital anomalies which has participated in EUROCAT Switzerland since 1988. During the period 1980-1996, we found 240 new T21 cases, all cytogenetically proven, out of 115,064 consecutive live births. Our purpose was to study trends and impact of biochemical screening and prenatal diagnosis of T21. We considered two different periods: 1980-1989 (before biochemical screening) and 1990-1996 (with screening) during which the mean maternal ages were respectively 28.4 years (10.6% > or = 35) and 29.2 years (12.9% > or = 35). The total prevalence of T21 was 2.08 per 1000; 5.4% of the cases were stillbirths, 49.6% were induced abortions and 45% livebirths. Prenatal cytogenetic diagnosis of trisomy 21 was performed in 52.1% of cases. Among women aged 35 or over the prenatal detection rates are superposable in the two periods. However, for younger women this rate has been much higher since the introduction of biochemical screening, i.e. 9.8% before and 51.8% after the introduction of triple test. In conclusion, the increase in prenatal diagnosis tests performed because of abnormal maternal serum marker levels has increased the global prenatal detection rate from 36.6% to 63.3% in our population, and the prevalence of Down syndrome has thus slightly decreased among livebirths. PMID- 11064926 TI - [Turner syndrome]. AB - This article is based on the study of 52 cases of Turner's syndrome, born between 1980 and 1996 and recorded in the Registry of Congenital Anomalies in the Canton of Vaud. In most cases the cytogenetic analysis was based on maternal multiple marker screening, sonography findings or maternal age. The most common chromosome abnormality is complete monosomy X. The rare cases of mosaic and the one case of isochromosome mainly involve livebirths. Morphological analysis of foetuses revealed hygroma colli (84%) and hydrops (63%), frequently associated with major cardiac malformations. The livebirths present growth retardation, pterygium colli and facial dysmorphic features, but rarely complex malformations. In the light of our data, the probability of survival to birth is 0.8% and the prevalence in all clinical pregnancies is 1.1%. PMID- 11064927 TI - [A rare case of aortic regurgitation]. PMID- 11064928 TI - [Ten years of Longitudinal Aging Study Amsterdam. A special issue]. PMID- 11064929 TI - [Longitudinal research in aging populations: participation and the quality of data collected with questionnaires]. AB - In this article methods are discussed which may keep participation in a longitudinal study among elderly persons high, for example adaptation of the interviews and proxy interviews. The LASA sample is described from the start in 1992 until now. The non-response is evaluated and we found that refusals are particularly important in the first part of the longitudinal traject. Also data quality is studied in relation to the aging of the respondents. Although there are theoretical reasons to expect that aging may impair data quality, no support for this hypothesis was found in the present study. Data quality was stable during a period of six years. But data quality seemed poorer for those respondents who dropped out from the study. Item non-response and duration of the interview were higher for drop-outs. PMID- 11064930 TI - [Effect of changes in physical capacities upon personal social network in aging people]. AB - The aim of the research is to assess whether there is change in the size and composition of older adults' personal network. Furthermore, change in contact frequency and received instrumental support within the relationships is studied. Five relationship types are distinguished: children, other kin, friends, neighbors and acquaintances. Older adults with a decline in physical capacity are compared with those with stable and increased capacities. Furthermore, differences according to (change in) partner status and age are investigated. Data are from the Longitudinal Aging Study Amsterdam, including the first and fourth observation of 1634 older adults living independently. The observation interval is 7 years. A decline in physical capacities is observed for 35% of the older adults, the capacities are stable for 60% and an increase is observed for 5% of the respondents. In general, network size and composition did not change. The frequency of contact within the relationships decreased. Decline was considerable for parent-child relationships, but relatively modest among older adults who faced a moderate to strong physical decline. However, among older adults who did not have a partner at the fourth observation and among the oldest (> 75 years) the frequency of contact with children increased, independently of the degree of physical decline. The decline in contact with neighbours was nearly absent for older adults who faced a moderate to strong physical decline; the contact increased when there was no partner at the fourth observation. The instrumental support received increased within all relationship types, independently of the degree of physical decline. It is concluded that research into determinants of the decline and increase in parent-child contacts is needed and that the meaning of neighbours should receive attention. PMID- 11064931 TI - [Apolipoprotein E4 and memory decline in the elderly]. AB - The objective of this study was to investigate whether the association between Apolipoprotein E4 (ApoE4) and memory decline is modified by baseline general cognitive impairment and age in a population-based elderly sample. Subjects were participants in the Longitudinal Aging Study Amsterdam (LASA). The study sample consisted of 1,243 subjects, 62-85 years old, with a Mini-Mental State Examination (MMSE) score between 21-30 and known ApoE phenotypes. Memory performance was measured with a short version of the Auditory Verbal Learning Test (AVLT) at baseline and repeated after three years (N = 854). Memory decline was defined as a decrease of at least one standard deviation from the mean change score on immediate recall, delayed recall and retention. ApoE4 was associated with memory decline in cognitively impaired subjects (MMSE 21-26), but not in cognitively normal subjects (MMSE 27-30). In particular cognitively impaired E4 carriers older than 75 years were at high risk of memory decline. Contrary to AD studies, our study suggests that the risk of ApoE4 on memory decline does not decrease with ageing. PMID- 11064932 TI - [Health status and anxiety in the elderly. A longitudinal perspective]. AB - The prognostic value of physical health for changes in anxiety symptoms in older people was investigated in a prospective longitudinal study design with data from the Longitudinal Aging Study Amsterdam (LASA). In a sample of 2165 older (> 55 yrs.) respondents anxiety symptoms were measured twice over a three year interval with the anxiety subscale of the Hospital Anxiety and Depression Scale (HADS-A). Utilizing a cut-off value of 4 on the HADS-A, subjects were considered as anxious or as non-anxious. Based on the first assessment two groups were formed: subjects with and subjects without anxiety symptoms. In the non-anxious cohort the effect of physical health on the development of anxiety symptoms was studied; in the anxious cohort the same factors were evaluated on their predictive value for chronicity of anxiety. Indices of physical health included the presence of chronic diseases, functional limitations, and self-perceived health at the first assessment and changes on these variables over time. Results revealed that poor self-perceived health was predictive of incidence (OR = 1.5; 95% CI = 1.3-1.8) and chronicity of anxiety (OR = 1.2; CI = 1.0-1.5). Regarding chronic diseases, the results showed that suffering from more than one chronic disease predicted becoming anxious and chronicity of anxiety (OR = 1.7; CI = 1.2-2.5 and OR = 2.2; CI = 1.3-3.6, respectively). Specific chronic diseases were not strongly related to a change in anxiety levels. Thus, somatic diseases not only lead to depression, a finding reported in numerous studies, but also increase the likelihood of anxiety symptoms at a later point in time. PMID- 11064933 TI - [Effects of depression on physical health and mortality in the elderly. Longitudinal results of the LASA research]. AB - This longitudinal study examines the physical health consequences of depression among 3107 older persons (55-85 years). Major depression was defined according to DSM-III criteria in a psychiatric interview. Minor depression was defined by a Center for Epidemiologic Studies Depression score > or = 16. Health consequences were assessed by 3-year change in self-reported functional status, 3-year change in performance on objective tests, and risk of death over 4.5 years. At baseline, 12.8% of the older persons had minor depression and 2.0% major depression. Minor depression was associated with a significantly greater decline in functional status and performance and, only in men but not in women, with an increased risk of death. Major depression also increases decline in functional status and the risk of death (irrespective of sex), but was not associated with decline in physical performance. These results show that late-life depression has strong unfavorable physical health consequences. The consequences of minor depression are comparable with those of major depression. PMID- 11064934 TI - [Social-economic inequality in mortality among older mem amd women. Research on the role of health, life style, parental socio-economic status and psychosocial conditions]. AB - This article describes to what degree socio-economic differences exist among community living older men and women, and to what degree these differences are to be explained by health, behaviour, childhood and psychosocial conditions. The data are available from 1427 men and 1503 women (aged 55-85), participating in the Longitudinal Aging Study Amsterdam (LASA) in 1992/1993. As indicators of socio-economic status (ses) we used the highest level of education and net monthly income. Age-adjusted mortality risks for men and women with low income and for men with a low level of education are about 1.5 times as high as for to the persons with high income and educational level. Among men, but not among women, the difference in mortality risk between low and high status persons remains after adjustment for age, health status, and several risk factors. Differences in lifestyle, parental ses and psychosocial characteristics explain little to nothing of the age-adjusted ses-differentiation in mortality. It is concluded that ses-inequalities in mortality are present among Dutch men and, to a lesser extent among women, until high age, and are partly explained by the relatively large health problems of the lower status group. PMID- 11064935 TI - Efficient sample pre-concentration of bupivacaine from human plasma by solid phase extraction on molecularly imprinted polymers. AB - The ability to use imprinted polymers for solid-phase extraction is demonstrated in a model pre-concentration of bupivacaine from human plasma samples prior to gas chromatography. Imprinting of the structural analogue pentycaine yielded a sorbent which efficiently extracted analyte and internal standard, while possible interference on analyte quantification from leakage of remaining template molecules was eliminated. Human plasma samples were diluted with citrate buffer pH 5, and applied onto solid phase extraction columns containing 15 mg of imprinted sorbent. Wash steps with 20% methanol in water followed by acetonitrile preceded elution with 2% triethylamine in acetonitrile. A direct comparison with conventional sample pre-treatment methods showed the high selectivity of the imprinted sorbent resulted in distinctly cleaner chromatographic traces than were obtained both after liquid-liquid extraction and C18-based solid-phase extraction. PMID- 11064936 TI - Trichosanthin induced calcium-dependent generation of reactive oxygen species in human choriocarcinoma cells. AB - The type-I ribosome-inactivating protein trichosanthin (TCS) has a broad spectrum of biological and pharmacological activities, including abortifacient, anti-tumor and anti-HIV. We found for the first time that TCS induced the generation of reactive oxygen species (ROS) in human choriocarcinoma cells (JAR cells) at the level of the single cell by using the fluorescent probe 2',7'-dichlorofluorescein diacetate with confocal laser scanning microscopy. TCS-induced ROS formation was shown to be dependent on the presence of extracellular Ca2+ and was further reduced when cytosolic Ca2+ was chelated by BAPTA-AM. The production of ROS increased rapidly after the application of TCS, which paralleled TCS-induced increase in intracellular calcium monitored using fluo 3-AM. Simultaneous observation of the nuclear morphological changes via two-photon laser scanning microscopy and production of ROS via confocal laser scanning microscopy revealed that ROS were involved in the apoptosis of JAR cells. The contribution of ROS was confirmed by experiments in which the antioxidant alpha-tocopherol prevented TCS induced ROS formation and cell death. The finding that TCS induced calcium dependent generation of ROS in JAR cells and that ROS were involved in the apoptosis of JAR cells might provide new insight into the anti-tumor and anti-HIV mechanism of TCS. PMID- 11064937 TI - Clean-up, detection and determination of salbutamol in human urine and serum. AB - Salbutamol ?2-(tert-butylamino)-1-[4-hydroxy-3- (hydroxymethyl)phenyl]ethanol?, also known as albuterol, is clinically the most widely used beta 2-adrenoceptor agonist in the treatment of bronchial asthma. During this study, we evaluated liquid-liquid extraction (LLE) and solid-phase extraction (SPE) in order to develop a reliable extraction method followed by analysis using liquid chromatography and gas chromatography. An assay is described which involves SPE as the clean-up method followed by gas chromatography-mass spectrometry to determine salbutamol levels in human serum after oral administration. The SPE method requires the use of a hyper-cross-linked styrene-divinylbenzene bonded phase (ENV+) without involving any sample pre-treatment to obtain 60-65% recoveries for salbutamol and terbutaline as the internal standard. Distilled water and 1% trifluoroacetic acid in methanol were found to be the most suitable washing solvent and eluting solvent, respectively. A detection limit of 2 ng mL-1 was achieved by derivatization with N-methyl-N-trimethylsilyltrifluoroacetamide to form trimethylsilyl (TMS)-salbutamol (m/z 369) and TMS-terbutaline (m/z 356). The relationship between the ratio of the peak area of salbutamol to that of the internal standard and concentration was linear for the range tested (2-200 ng mL 1) and the correlation of coefficient was 0.9999 with a y-intercept not significantly different from zero. The inter-day relative standard deviation (RSD) was < 10% for all three concentrations. The intra-day RSD was 14% for 2 ng mL-1. This assay was then successfully applied to human serum samples obtained from clinical trials after oral administration of salbutamol. PMID- 11064938 TI - An improved assay of 3-hydroxy-3-methylglutaryl-CoA reductase activity in Reuber H35 hepatoma cells without microsomes isolation. AB - The optimal conditions for measuring 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase activity in Reuber H35 hepatoma cells are described in this paper. Cells in the exponential phase of growth were lysed by incubation with Brij 97 detergent for 30 min. We used imidazole buffer supplemented with EDTA and leupeptine, two inhibitors of proteases. Disrupted cells were then centrifuged at 12,000 g. Although microsomes are usually reported as enzyme preparations for measuring HMG-CoA reductase, our data showed that hepatoma cells may be used without previous isolation of microsomes. The 12,000 g supernatant showed similar levels of total and specific activities to those found in the microsomal fraction obtained after 105,000 g centrifugation. The soluble fraction showed less than 10% of reductase activity. Reductase activity from Reuber H35 hepatoma cells increased proportionally to the reaction time from 30 to 90 min and to the amount of protein added in a range of 50-500 micrograms. Our modified method was very sensitive and reproducible, because very low specific activity (about 15-100 pmol min-1 per mg protein) could be quantified in different assay conditions obtaining similar values. PMID- 11064939 TI - Renewable amperometric immunosensor based on paraffin-graphite-transferrin antiserum biocomposite for transferrin assay. AB - A renewable electrochemical immunosensor was developed for the determination of transferrin in human serum. It is based on a paraffin-graphite-transferrin antiserum biocomposite, which needs no additional curing. A competitive binding assay was used to determine transferrin in human serum with the aid of transferrin labeled with horseradish peroxidase. The assay conditions were optimized, including the loading of transferrin antiserum in the biocomposite, the amount of labeled transferrin in the incubation solution, incubation time and temperature. Serum samples were analyzed and the results demonstrate that the concentration range of determination with this system meets the demands of clinical analysis. The surface of the immunosensor can be regenerated by simply polishing to obtain a fresh immunocomposite ready to be used in a new competitive assay. PMID- 11064940 TI - Adsorptive stripping voltammetric determination of ketoconazole in pharmaceutical preparations and urine using carbon paste electrodes. AB - The oxidation of ketoconazole on a bare carbon paste electrode was studied voltammetrically. The results indicated that the process is irreversible and controlled by an adsorption-extraction process which allows the accumulation of the drug at the electrode surface. After the optimization of solution pH, accumulation variables and instrumental parameters, sensitive differential pulse and linear sweep voltammetric peaks were obtained whose peak currents were linearly proportional to the ketoconazole concentration over the ranges 2.4 x 10( 8)-4.8 x 10(-7) and 9.1 x 10(-7)-1.0 x 10(-5) M, respectively. Based on these findings, a simple procedure was developed for the determination of ketoconazole in human urine and formulations. PMID- 11064941 TI - Estimation of trace amounts of benzene in solvent-extracted vegetable oils and oil seed cakes. AB - A new method is presented for the qualitative and quantitative estimation of trace amounts (up to 0.15 ppm) of benzene in crude as well as refined vegetable oils obtained by extraction with food grade hexane (FGH), and in the oil seed cakes left after extraction. The method involves the selection of two solvents; cyclohexanol, for thinning of viscous vegetable oil, and heptane, for azeotroping out trace benzene as a concentrate from the resulting mixture. Benzene is then estimated in the resulting azeotrope either by UV spectroscopy or by GC-MS subject to availability and cost effectiveness of the latter. Repeatability and reproducibility of the method is within 1-3% error. This method is suitable for estimating benzene in vegetable oils and oil seed cakes. PMID- 11064942 TI - Drugs for the doctor's bag revisited. AB - The choice of drugs to include in the GP's bag depends on the medical conditions likely to be met, the shelf-life of the products and their costs, the availability of ambulance paramedic cover and the proximity of the nearest hospital. Here, we update previous advice and suggest a list of medicines that GPs may wish to take with them on home visits for use in an emergency or other acute treatment. We include paediatric doses where appropriate and, whenever a medicine is first mentioned, our suggested formulation is given italicised and in brackets. We also enclose with this issue a card summarising parenteral doses of drugs for medical emergencies, which includes a table of mean weight for age. PMID- 11064943 TI - Managing falls in older people. AB - Falls are a common problem in older people. Each year in the community, among those aged 65 years or over, around one-third fall at least once and about 8% on average experience a fall with injury that receives acute medical attention. Here, we consider the management of older people who fall. PMID- 11064944 TI - Post-prandial hyperglycaemia--the necessity for better control. PMID- 11064945 TI - Clinical significance of post-prandial blood glucose excursions in type 1 and type 2 diabetes mellitus. PMID- 11064946 TI - Post-prandial hyperglycaemia and vascular disease. PMID- 11064947 TI - The role of the liver on post-prandial glycaemia--emphasis on hepatic glucose uptake. PMID- 11064948 TI - Improving post-prandial control with Humalog and Humalog mixtures. PMID- 11064949 TI - Hypoglycaemia awareness and counter-regulation in type 1 diabetes. PMID- 11064950 TI - Adaptations leading to hypoglycaemia in type 1 diabetes mellitus and comparison with type 2 diabetes mellitus. PMID- 11064951 TI - Incidence and predisposing factors of hypoglycaemia in children and adolescents with diabetes. PMID- 11064952 TI - Hypoglycaemia--clinical consequences and morbidity. PMID- 11064953 TI - Hypoglycaemia as a barrier to improved glycaemic control. PMID- 11064954 TI - Experiments on the meaning of four types of single-accent intonation patterns in Dutch. AB - The Grammar of Dutch Intonation (GDI) provides a description of the possible intonation contours of Dutch. The GDI distinguishes accent-lending and nonaccent lending pitch configurations, but refrains from further functional statements. This paper describes an experimental attempt to verify meaning hypotheses for four Dutch single-accent pitch patterns as postulated in the linguistic literature. The four pitch accent types were realized on proper names; the abstract meanings, in terms of the manipulation of an element of the background shared between speaker and listener, were incorporated in situational contexts, distinguishing between a "default" and a vocative use of the proper name ("orientation"). Listeners ranked the four melodic shapes from most to least appropriate in their specific context. After revision of part of the materials a second perception experiment was conducted, in which subjects had a rank four contexts from most to least appropriate for a specific pitch accent type. Results show a distinct effect of "orientation" on the appropriateness of two of the investigated pitch accent types in the various context types; the other two pitch accent types are associated with the predicted context types (and vice versa) well above chance, indicating the viability of at least two of the linguistic proposals. PMID- 11064955 TI - The perception of lexical tone in Mambila. AB - The issue of the perception of lexical tone has been addressed mainly through studies of Southeast Asian languages which feature phonological contour tones as well as level tones. Little attention has been paid to African languages which have, almost exclusively, only level tones. This paper examines tone perception in Mambila, a Benue-Congo language with four level lexical tones. A categorization experiment was run to determine some of the salient aspects of the perceptual nature of these tones. Since the four tones are well defined with respect to production, we sought to determine whether this characteristic carried over into perception, the expectation being that experimental stimuli, on the basis of pitch height alone, would fall into four reasonably well defined categories. Results showed interesting differences across the four tones, with indications that the two Mid tones, T2 and T3, are perceptually different than the High (T1) and Low (T4) tones. The experiment was run a second time, using a group of native English listeners, to assess to what extent results for the Mambila listeners were determined by the perceptual structure of the Mambila tone system. A Signal Detection analysis was used, which revealed important differences between the two groups of listeners. Results are discussed in light of what is known about universal tendencies of tone systems and the historical development of the Mambila system. PMID- 11064956 TI - The behavior of H* and L* under variations in pitch range in Dutch rising contours. AB - A Dutch rising intonation contour can be realized either as a rise that begins low and ends mid-to-high ("low rise") or as a rise that begins mid and ends high ("high rise"). These two contours could either be the extremes on a phonetic continuum representing a single phonological contour, for instance L* H-H%, or be realizations of two phonologically different contours, L* H-H% and H* H-H%. In order to decide between these two analyses, listeners were asked to rate stimuli with different pitch ranges on a number of semantic scales whose meanings vary with pitch range. Our hypothesis was that H-tones are higher as the pitch range increases, while L* is lower. Two preliminary experiments, in which we presented F0 contours of high rises and low rises in a number of different pitch ranges, revealed that perceived surprise, rather than perceived prominence, is an appropriate response variable for measuring pitch range perception, where increased pitch range corresponds to higher H-tones and lower L*. Subsequently, listeners were asked to indicate the degree to which each of a number of appropriately manipulated stimuli expressed "Surprise." The results lend strong support to the hypothesis that the low rise and the high rise are categorically distinct contours of Dutch, and that their first tones are L* and H*, respectively. PMID- 11064957 TI - Learning long words--a typological perspective. AB - The early phonological development of six normally developing children learning Finnish is reported. The primary focus is on their production of multisyllabic targets. The one-syllable and two-syllable forms found in the speech of children learning English has often been seen as a universal feature of early words. Later research has indicated that, for example, some children learning Japanese may produce word forms that contain three to five syllables already at the age of 15 months among the first fifty words. In Finnish long words are common and as expected, children targeted them already at the first word stage. However, they succeeded only after this stage had passed. Individual differences were extensive. Although the main tendency seems to reduce the last element of long words producing the SW1-pattern, important exceptions can be found so that the results do not provide strong support for a metrical template. The study also indicated that the segmental factor may influence the deletion pattern: the syllable that contained a stop was produced, regardless of its position. PMID- 11064958 TI - Family experiences and factors associated with the diagnosis of fragile X syndrome. AB - The authors interviewed 41 mothers of young boys with fragile X syndrome to determine the process by which they learned their child had fragile X syndrome. The average family had concerns about the child's development at 9 months of age. Developmental delay was determined at an average age of 24 months, and fragile X syndrome was diagnosed at a mean age of 35 months. Considerable variability was found in age of first concern, determination of delay, and diagnosis of fragile X syndrome. Three child variables (severity of delay, autistic behavior, temperament style) and four family variables (mother's age, mother's education, sibling status, social support) did not account for this variability, although birth year did (children born more recently were somewhat more likely to be identified earlier). Families often encountered physicians who initially discounted concerns or said that it was too early to determine whether a problem did indeed exist. Given current knowledge and practice, improving the early identification (under 3 years of age) of children with fragile X syndrome is likely to remain difficult if based solely on behavioral and clinical observations. PMID- 11064959 TI - Medication management of stimulants in pediatric practice settings: a national perspective. AB - Using a nationally representative sample of office-based physicians, the management practices of pediatricians, psychiatrists, and family practice physicians were investigated. The major aims were to determine (1) what types of services these physicians were providing to children who received stimulants, (2) what factors predicted receipt of stimulants, and (3) whether these practices were concordant or discordant with professional consensus on diagnosis and treatment of attention-deficit hyperactivity disorders (ADHD). Prescribing and management practice data from the 1995 National Ambulatory Medical Care Survey (NAMCS) were analyzed for children ages 0 to 17 years who were seen for psychiatric problems and received stimulant medication. Results indicated that 2 million visits by children were made in 1995 to psychiatrists, pediatricians, or family practitioners in which psychotropic medications were prescribed. In pediatric visits where stimulant medication was prescribed, mental health counseling was provided 47.3% of the time and psychotherapy 21.6%. Follow-up arrangements were made in 79.1% of the visits. Psychiatrists were significantly more likely to provide psychotherapy and to specify follow-up visits than were pediatricians, but less likely to provide other health counseling. Controlling for demographic and physician effects, the factors with the most significant effect on the probability of receiving stimulants were geographic region (living in the South), race (being white), receiving mental health counseling, not receiving psychotherapy, and having health insurance. Less than 50% of pediatric visits for psychiatric reasons involving stimulant medications included any form of psychosocial intervention. In 21% of these visits, no recommendations were made for follow-up care. These practices diverge from National Institutes of Health (NIH) consensus panel recommendations and association-issued practice parameters. PMID- 11064960 TI - Family reinforcement of illness behavior: a comparison of adolescents with chronic fatigue syndrome, juvenile arthritis, and healthy controls. AB - Parental encouragement of illness behavior is hypothesized to correlate with psychosocial dysfunction in adolescents with chronic illness. To explore this hypothesis, adolescents aged 11 to 17 years with chronic fatigue syndrome (CFS) (n = 10), juvenile rheumatoid arthritis (JRA) (n = 16), and healthy adolescents (n = 14) were recruited for the study. Measures included the Achenbach parent and youth self report forms, the Family Adaptability and Cohesion Evaluation Scale-II (FACES II), the Children's Depression Rating Scale, and number of days absent from school. The Illness Behavior Encouragement Scale (IBES) generated measures of parental reinforcement of illness behavior. As predicted, the teens with CFS scored statistically higher on measures of depression, total competence, and number of days of school missed in the previous 6 months (mean = 40). Children with JRA scored significantly lower than the CFS group on the measure of parental reinforcement of illness behavior. The healthy group produced intermediate scores. Results and implications for future clinical and research activity are discussed. PMID- 11064961 TI - Cognitive-behavioral predictors of asthma morbidity in inner-city children. AB - Asthma is a growing health problem among children in the United States, particularly in urban, inner-city areas. This article examines the relationship between cognitive-behavioral aspects of asthma management (caretaker asthma knowledge, expectations, and problem-solving) and asthma morbidity in a sample of 1,376 inner-city children with physician-diagnosed asthma. In the analyses, baseline symptom severity served as a covariate, and the average of the 3-, 6-, and 9-month follow-up data served as the outcome measure. Children of caregivers with ineffective problem-solving strategies had significantly more days of wheezing over a 14-day period. Ineffective problem-solving capabilities were also associated with poorer functional status; however, positive caregiver expectations were associated with better functional status. Of the cognitive behavioral factors studied in a high-risk urban population, caregiver problem solving skills and expectations emerged as meriting further investigation and possible intervention. PMID- 11064962 TI - Placebo effects in autism: lessons from secretin. PMID- 11064963 TI - Feeding problems, sleep disturbances, and negative behaviors in a toddler. AB - Tiffany, a 3-year-old girl, was referred to the developmental and behavioral pediatrics service for evaluation of significant and persistent negative behaviors associated with refusal to eat at meal time and constant snacking during the past 3 months. She lost 2 pounds, but her weight for her height was at the 50th percentile. Her mother indicated that Tiffany had frequent night awakenings (>10) and late sleep onset (between 12:00 and 1:00 a.m.). Her mother described her as being "easily frustrated," getting upset and angry very quickly. Tiffany was identified at an early intervention program as having mild to moderate developmental delays in pragmatic speech, gross and fine motor skills, and social interaction skills. Tiffany was born at 33 weeks gestation and was hospitalized for 10 days without significant perinatal problems. She was readmitted at 2 months of age when she was diagnosed with gastroesophageal reflux, lactose intolerance, sleep apnea, and bradycardia. She was discharged with an apnea monitor. A seizure disorder was diagnosed at 1 year of age and reactive airway disease at 2 years of age. At the time of the referral to the developmental and behavioral pediatrics service, Tiffany was followed by multiple services, including cardiology, neurology, gastroenterology, psychology, and pulmonary. Pharmacologic therapies included albuterol and cromalyn inhalers, phenobarbital, valproic acid, levocarnitine, ranitidine, and an inhaled steroid. She continued to use the apnea monitor each night, although three sleep studies demonstrated a normal sleep pattern with no evidence of apnea or bradycardia. A recent electroencephalogram was normal. Tiffany lives with her mother and maternal grandparents. Her mother is morbidly obese with a history of asthma and depression. She was infertile for a 10-year period, which she attributed to the stress associated with living with an abusive man. Tiffany was the result of a subsequent, brief relationship with another man; she has not had contact with her father. Her mother is a licensed practical nurse who has not worked as a nurse since Tiffany's birth. An interdisciplinary treatment approach to Tiffany's multiple biological and behavioral problems was implemented by admitting her to a collaborative care unit at a children's hospital. PMID- 11064964 TI - Behavior problems and group-based parent education programs. AB - Behavior problems in children are an important social, educational, and health issue. The prevalence of these problems, their stability over time, their poor prognosis, and their costs to both individuals and the society, all point to the need for primary prevention and early effective interventions. A systematic review examined the effectiveness of group parent education programs that aimed to improve behavior problems in 3- to 10-year-old children. The phrase "parent education program" is used here to refer to group-based programs with a standardized format aimed at enhancing parenting skills. The term "behavior problems" is used to refer to children exhibiting externalizing problems such as temper tantrums, aggression, and noncompliance. It does not include children diagnosed as having attention-deficit hyperactivity disorder. This review focused explicitly on measures of child behavioral outcomes, which are only small, albeit important, outcomes of parent education programs. Reviews focusing on other clinically relevant outcomes are also needed, including parental well-being and attitudes towards parenting. Other reviews are also needed to collate evidence concerning the effectiveness of parent education programs with other age-groups, i.e., preschoolers and adolescents, and in improving other aspects of child well being. The review included published studies only and as such may have been influenced by a "publication bias." Inclusion criteria comprised the use of a waiting list, a no-treatment or placebo control group, and at least one standardized measure assessing the child's behavior. Only studies published after 1970 that included at least one "group-based" parent education program were included. A total of 255 primary studies were identified, but only 16 of these and 2 follow-up studies met all of the specified inclusion criteria. Critical appraisal of these 16 studies revealed considerable heterogeneity in the interventions, the populations studied, and the outcome measures used. Nevertheless, these studies suggest that structured parent education programs can be effective in producing positive change in both parental perceptions and objective measures of children's behavior and that these changes are maintained over time. Because of the small number of controlled studies and their methodological variations, caution should be exercised before these findings are generalized broadly. PMID- 11064965 TI - The escalating use of psychotropic medications to treat behavior problems in preschoolers. PMID- 11064966 TI - Presidential address: developmental-behavioral pediatrics and primary care. PMID- 11064967 TI - Geometric total knee replacement: operative considerations. Lee Hunter Riley, Jr., MD 1973. PMID- 11064968 TI - Articulating versus static spacers in revision total knee arthroplasty for sepsis. The Ranawat Award. AB - Antibiotic laden spacer blocks frequently are used to treat an infected total knee arthroplasty. Static spacer blocks make exposure at reimplantation difficult secondary to quadriceps shortening. Unexpected bone loss attributable to migration of the spacer block also has been reported. To avoid these problems, a temporary articulating molded implant made of antibiotic cement was used in a consecutive series. The authors sought to determine whether its use would affect the reinfection rate, improve functional results, or prevent bone loss compared with static spacers. Twenty-five patients were treated with static nonarticulating spacers. Since 1996, 30 patients have been treated with tobramycin-laden articulating spacers. The knee arthroplasties in three patients treated with a static spacer became reinfected (12%). The knee arthroplasty in one patient with an articulating spacer became reinfected (7%). Fifteen of the 25 patients with static spacers had unexpected bone loss between stages. No appreciable bone loss could be measured in the patients who received articulating spacers. The average Hospital for Special Surgery score was 83 points in the patients with static spacers and 84 points for the patients with articulating spacers. Range of motion at final followup averaged 98 degrees in the patients who received static spacers and 105 degrees in the patients who received articulating spacers. Articulating spacers seem to facilitate reimplantation of infected total knee arthroplasty without additional risk of infection. Unexpected bone loss is no longer a concern with this two-stage technique. Articulating spacers offered no functional advantage over static spacers in this study group. PMID- 11064969 TI - Ten- to 12-year followup of the Insall-Burstein I total knee prosthesis. AB - The Insall-Burstein Posterior Stabilized knee prosthesis (Insall-Burstein I), developed at The Hospital for Special Surgery in 1978, has a metal-backed nonmodular tibial component. The polyethylene articular surface was directly molded. The purpose of the current study was to evaluate long-term wear with this design. The first 100 total knee arthroplasties (86 patients) performed by the senior author were followed prospectively. The average age of the patients at the time of surgery was 69.7 years (range, 45-89 years). The primary diagnoses were osteoarthritis in 77 knees (66 patients), inflammatory arthritis in 17 knees (14 patients), and posttraumatic arthritis in the remaining six knees (six patients). Thirty-eight knees (35 patients) had varus angulation, 14 knees (13 patients) had valgus angulation, and 48 knees (40 patients) had a 0 degrees to 10 degrees tibiofemoral angle preoperatively. All patients were evaluated at 10 to 12 years followup. Knee Society scores and radiographs were obtained. Thirty-six knees were in 30 patients who had died and two knees were in two patients who were infirm. Telephone evaluation only was available for eight knees (seven patients), leaving 54 knees (47 patients) for direct clinical and radiographic evaluation. No patients were lost to followup. The average Knee Society clinical score at latest followup was 91.6 points. The average function score was 69 points. One knee arthroplasty failed because of tibial loosening, one failed because of patella wear and fracture, two failed because of sepsis, and two failed because of nonspecific pain. There were seven patella fractures (7%) in the 100 knees. One of the fractures resulted in a total knee revision (noted above), two resulted in patellar component revision, and another resulted in patellar component removal. The remaining three patella fractures were discovered incidentally and were asymptomatic. There were no patellar dislocations. At long term radiographic analysis, valgus alignment averaged 6 degrees (range, 0 degrees 11 degrees). Polyethylene wear averaged 0.40 mm. There was no catastrophic wear of tibial polyethylene. Thirty-two knees in 32 patients (65%) had radiolucencies in at least one zone; no lucency filled a zone, and none was wider than 2 mm. The absence of clinically significant tibial polyethylene wear at long-term followup is of particular interest. The performance of the molded, nonmodular polyethylene articulation is encouraging and needs to be analyzed critically against the more widely used machined, modular components used today. PMID- 11064970 TI - Prospective randomized clinical trial of continuous passive motion after total knee arthroplasty. AB - The authors report the results of a prospective randomized clinical trial using continuous passive motion after total knee arthroplasty. One hundred twenty patients were assigned randomly to one treatment group: No continuous passive motion (Group I), continuous passive motion from 0 degrees to 50 degrees and increased as tolerated (Group II), and continuous passive motion from 70 degrees to 110 degrees (Group III). The continuous passive motion was initiated in the recovery room and was maintained for a maximum of 24 hours at which point all patients began identical postoperative physiotherapy regimens. Patients were assessed preoperatively, during their hospital stay, at 6 weeks, 12 weeks, 26 weeks, and 52 weeks after their surgery. There were no statistical differences between any of the treatment groups regarding cumulative analgesic requirements, range of motion at any measured interval, length of stay (Group I, 5.1 days; Group II, 5.2 days; Group III, 5 days) or Knee Society scores. The current study does not support the use of short-term continuous passive motion after total knee replacement. A standard and a high flexion continuous passive motion protocol failed to show any advantage over physiotherapy alone in the parameters evaluated. PMID- 11064971 TI - Deep-dish congruent tibial component use in total knee arthroplasty: a randomized prospective study. AB - One hundred seventy-six patients with osteoarthritis of the knee were randomized prospectively into two groups. In both groups the posterior cruciate ligament was released from its femoral attachment. In one group a posterior stabilized tibial component was used whereas in the other group a deep-dish tibial polyethylene component was inserted (Genesis II). The surgical and perioperative technique was identical in both groups and all the implants were cemented to their respective bones. Patients began range of motion exercises within the first few hours after surgery and were allowed weightbearing to tolerance beginning on the first postoperative day. At followup there was no statistical difference in the mean range of flexion (approximately 116 degrees), ability to ascend and descend stairs in a bipedal manner (80%), pain scores, knee scores (94 points), stability, or the lack of anterior knee pain. Postoperative implant alignment in the sagittal and coronal planes and on Merchant skyline views was excellent in both groups. There was only one lateral release required and that was in one patient who received a deep-dish component. Using deep-dish implant obviates the need to resect intercondylar femoral bone, decreasing the potential for fracture and maximizing bone volume should revision be necessary in the future. PMID- 11064972 TI - Functional medical ligament balancing in total knee arthroplasty. AB - Function of the anterior and posterior oblique portions of the medial collateral ligament and the posterior capsule in flexion and extension was evaluated in eight knee specimens after posterior cruciate retaining total knee arthroplasty. The posterior oblique portion of the medial collateral ligament was released subperiosteally in four specimens, and the anterior portion was released in four specimens. The medial posterior capsule was released in each group, then the remaining portion of the medial collateral ligament was released. Release of the posterior oblique portion produced moderate laxity at full extension and at 30 degrees flexion, and posterior capsule release produced additional laxity in full extension. Release of the anterior portion produced major laxity at 60 degrees and 90 degrees flexion. Complete medial collateral ligament release increased laxity significantly in both groups in flexion and extension. This rationale was tested in a clinical study of 82 knees (76 patients) in which 62 (76%) required medial collateral ligament release to correct varus deformity during posterior cruciate retaining total knee arthroplasty. Twenty-two knees (35.5%) were tight medially in extension only, and were corrected by releasing the posterior oblique portion. Thirty-one knees (50%) were tight medially in flexion only, and were corrected by releasing the anterior portion. Nine knees (14.5%) were tight medially in flexion and extension and required complete medial collateral ligament release, but three knees (4.8%) remained tight in extension and required medial posterior capsule release to correct flexion contracture and medial ligament contracture. Seventeen (27%) had partial posterior cruciate ligament release to correct excessive rollback of the femoral component on the tibial surface. PMID- 11064973 TI - Primary constrained condylar knee arthroplasty for the arthritic valgus knee. AB - The purpose of the current study was to review results of primary constrained condylar knee arthroplasty in elderly patients with genu valgum deformity. The hypotheses were: (1) constraint has no adverse effects in elderly patients; (2) treating deformity with a constrained condylar knee prosthesis in lieu of lateral ligament release avoids morbidity, particularly peroneal nerve palsy and flexion instability; and (3) press-fit noncemented stem extensions enhance fixation of the cemented core components and are not prone to loosening. Between 1988 and 1993, 44 consecutive primary Constrained Condylar Knee prostheses were implanted in 37 patients (average age, 72.7 years) with an average valgus angle of 17.6 degrees. Indications for the Constrained Condylar Knee implant were: elderly patients with genu valgum deformity and medial collateral ligament incompetence. Outcome was assessed prospectively using the Hospital for Special Surgery and Knee Society scoring systems; followup was by independent observer. Clinical and radiographic followup (average, 7.8 years) was available for 28 knees (26 patients). The Hospital for Special Surgery score improved from 52.2 to 89.6 points. The average Knee Society score and functional scores improved from 27.4 and 32.4 points to 95.2 and 67.2 points, respectively. At followup, the average alignment based on anteroposterior radiographs obtained with the patient weightbearing was 5.3 degrees. No radiographic loosening, prosthetic failures, peroneal nerve palsies, or flexion instability occurred. No failures occurred in the 11 patients (16 knees) who died before the latest followup. To the authors' knowledge, this is the largest reported series with the longest reported followup of patients with primary Constrained Condylar Knee prostheses. The use of the Constrained Condylar Knee prosthesis for elderly patients with low physical demands with genu valgum resulted in significant pain relief and improved function. PMID- 11064974 TI - Athletic activity after total knee arthroplasty. AB - Americans are aging, elderly Americans are more active, and the prevalence of total knee arthroplasty is increasing. Indications for knee replacement include pain, deformity, and a desire to improve function. When patients have knee replacement operations, frequently they increase their activities. It is important for patients with knee replacements to understand the impact of athletic activity on the outcome of knee replacements. Orthopaedic surgeons should educate patients regarding athletic activity after total knee arthroplasty. Considerations and risk factors for athletic activity after knee replacements include athletic activity before surgery, preoperative rehabilitation, surgical reconstruction, implant failure or fracture, implant fixation or loosening, and joint bearing surface wear. Anatomic reconstruction and compulsive postoperative rehabilitation with restoration of muscular control are important for optimum function after total knee arthroplasty. In general, patients with knee replacements are encouraged to participate in low-impact, low demand sports, and to avoid high-impact, high-demand sports. PMID- 11064975 TI - Rotational malalignment of the femoral component in total knee arthroplasty. AB - Ligamentous balancing is a crucial part of total knee arthroplasty. To ensure proper kinematics, balance must be achieved in flexion and extension. Failure to do so may result in limited range of motion, premature polyethylene wear, or patellofemoral tracking problems. Balancing in extension is dependent on the type and extent of correctional ligamentous release. Flexion balance is dependent on proper femoral rotation. There are two methods to determine femoral rotation. In the classic method, the knee is tensed in flexion after ligamentous release in extension. The anteroposterior cut then is made parallel to the cut tibial surface. Alternatively, the anteroposterior cut can be based off fixed femoral landmarks. The purpose of the current study was to determine the variance between balancing the flexion gap with the classic method versus the technique of using fixed femoral landmarks to determine rotation. One hundred consecutive posterior stabilized knee arthroplasties were performed using the classic method. The resected posterior condyles in each case were measured. The actual difference between the resected condyles using the classic method was compared with the calculated difference of resected bone using bony landmarks to determine rotation. A variance analysis then was performed. Compared with classically balanced knees, rotational errors of at least 3 degrees occurred in 45 % of patients when rotation was determined from fixed bony landmarks. These patients had trapezoidal rather than rectangular flexion gaps. Such errors may have implications regarding polyethylene wear, range of motion, and long-term clinical results. PMID- 11064976 TI - The role of bladder catheterization in total knee arthroplasty. AB - The use of a urinary bladder catheter in the perioperative period for patients undergoing total knee arthroplasty is controversial. In the current study, two bladder management protocols were studied. One group of patients had an indwelling catheter inserted into the bladder before total knee arthroplasty. The other group of patients was observed and treated for urinary retention as necessary. From 1993 to 1998, 652 patients undergoing primary, unilateral total knee arthroplasty were randomized by surgeon into two groups: one group underwent preoperative insertion of an indwelling bladder catheter (306 patients), and one group (346 patients) had a catheter inserted postoperatively as necessary. Sixty six percent (229 of 346) of these patients required catheterization (203 had indwelling catheters and 26 had intermittent straight catheters). A urinary tract infection developed in five patients (1.6%) in whom a catheter was inserted preoperatively. A urinary tract infection developed in six patients (1.7%) in whom a catheter was inserted if necessary. Five of these urinary tract infections developed in patients with delayed indwelling bladder catheters. A urinary tract infection did not develop in any patient in whom a straight catheter was inserted. There was no significant difference in the length of stay in the hospital between the two groups. The group in whom a catheter always was inserted generated $491 greater cost for total knee arthroplasty than patients in whom a catheter was inserted if necessary. PMID- 11064977 TI - Total knee arthroplasty in patients 40 years of age and younger with osteoarthritis. AB - The results of 32 total knee arthroplasties performed for osteoarthritis in 32 patients who were 40 years of age or younger are reviewed. At a mean followup of 7.9 years (minimum, 5 years), the Knee Society knee scores increased from an average of 47 to 88 points, and the function scores increased from 45 to 70 points. Overall, Knee Society knee scores were considered good or excellent in 82% of patients (26 knees) and fair or poor in 18% (six knees). Postoperative function scores were good or excellent in only 40% (13 knees). The average postoperative flexion arc was 110 degrees. If patients involved in worker's compensation cases are excluded from analysis, the results improved substantially, with range of motion averaging 113 degrees, and Knee Society knee scores and function scores averaging 92 points and 77 points, respectively. Excluding the five patients involved in workmen's compensation cases, knee scores were good or excellent in 91% of patients (25 knees) and function scores were good or excellent in 50% of patients (14 knees). Three revisions were performed for aseptic failure; one additional patient has radiographic evidence of tibial loosening, representing an aseptic failure rate of 12.5% at 8 years. Although slightly higher than observed in older patients, this failure rate still may be considered acceptable for this population of patients with severely affected knees who are not considered candidates for nonarthroplasty surgery. Despite a slightly higher tendency for aseptic failures in this group of patients, cemented total knee arthroplasty may provide some patients younger than 40 years of age with severe debilitating and recalcitrant osteoarthrosis, an important option with reasonable mid- and long-term results. PMID- 11064978 TI - Treatment of periprosthetic tibial fractures. AB - Periprosthetic fractures of the tibia are less commonly encountered and have received less attention than periprosthetic fractures of the patella and distal femur. In contrast with distal femoral fractures, tibial fractures frequently are encountered with loose implants and treatment often requires simultaneous revision knee surgery to address the loose prosthesis, the fracture, and any associated bone deficiencies. In some instances, fractures associated with well fixed and satisfactorily positioned knee components may be treated by traditional methods of operative or nonoperative fracture management. A classification system, which accounts for the anatomic location of the fracture, the status of prosthesis fixation, and timing of the fracture is helpful in description of the various fracture patterns and direction of the appropriate treatment approach. PMID- 11064979 TI - The cost of teaching total knee arthroplasty surgery to orthopaedic surgery residents. AB - The higher costs associated with teaching hospitals have received some attention in the literature. The objective of the current study was to determine the increase in resource consumption associated with resident education in knee arthroplasty surgery. Seventy-four patients who underwent primary total knee arthroplasty in the same hospital were studied (50 private practice and 24 teaching practice). Time in the operating room and medical severity of illness were noted. Hospital charges were used as a measure of resource consumption. In addition, length of stay and in-hospital consultations and complications were observed. Kruskall-Wallis, chi square, and stepwise multiple regression analysis were performed. The mean age of the patients was 68 years. Patients who underwent surgery at the teaching service had higher charges ($30,311 +/- $3,325 versus $23,116 +/- $3,341) and longer times in the operating room (190 +/- 19 minutes versus 145 +/- 29 minutes). These patients also had a trend toward more associated comorbid medical conditions (0.71 versus 0.42). Stepwise multiple regression analysis showed that teaching was the most important predictor of charges and operating room time. The results show a 22% increase in perioperative resource consumption for patients who underwent surgery at a teaching service. The measured increase in cost is significantly lower than what has been reported in other series (82%). At the teaching institution, the anesthesia and orthopaedic surgery residents work together on all cases and perform a significant percent of the procedures under direct supervision. The increased resource consumption observed in a teaching service is most likely attributable to the hands-on approach taken to train residents. PMID- 11064980 TI - A randomized comparison of all-polyethylene and metal-backed tibial components. AB - Clinical failures of all-polyethylene tibial components in total knee arthroplasty generally have been failures of design, not materials. The current study was designed to compare a modern congruent all-polyethylene tibial component with a metal-backed tibial component with the same articular design and geometry. All patients older than 60 years of age requiring total knee arthroplasty were randomized prospectively to receive either a cemented posterior cruciate ligament-retaining all-polyethylene component or a metal-backed tibial component with identical articular surfaces. All patients received identical cemented femoral and all-polyethylene patellar implants. The mean age of the patients was 69 years, and the mean American Society of Anesthesiology score was 3. The diagnosis was osteoarthritis in 92% of this population. Three hundred twenty-four total knee arthroplasties in 296 patients were performed; 213 joints (111 all-polyethylene tibias and 102 metal-backed tibias) with a minimum of 3 years followup (mean, 49 months) are reported. The preoperative Knee Society knee score in the group of patients who received an all-polyethylene tibial component was 38 points, improving to 84 points at latest followup, whereas in the group of patients who received a metal-backed tibial component, the score improved from 35 to 85 points. Functional scores increased from preoperative values of 56 to 74 points in the patients who received all-polyethylene tibial components, and 57 to 72 points in the patients who received metal-backed tibial components. Range of motion measured at latest followup averaged 106 degrees in patients who received an all-polyethylene tibial component and 107 degrees in the patients who received a metal-backed component, and postoperative tibiofemoral alignment averaged 6 degrees valgus for both groups. There were 13 reoperations for instability, patellofemoral problems, or deep infection, but none for aseptic loosening or wear in either group. These differences were not statistically significant, nor were any measures of patient satisfaction or clinical outcome between the two groups in this period. Total knee arthroplasty with a well-designed, contemporary congruent all-polyethylene tibial component functions equivalently to its metal backed tibial counterpart at 3- to 5-year followup in this patient population, and is less costly ($675). PMID- 11064981 TI - Revision total knee arthroplasty: current rationale and techniques for femoral component revision. AB - Revision total knee arthroplasty often requires solutions for the multiple complexities that exist on the femoral side, including the treatment of bone loss, component position, ligamentous laxity, and component stability. Surgical decisions regarding the proper use of bone grafts, component augmentation, proper axial and rotational alignment, and femoral stems must be addressed systematically to achieve consistently successful outcomes. A review of currently used femoral revision techniques and their rationale and a classification system of femoral deficiencies designed to guide the surgical decision making process are presented. PMID- 11064982 TI - Long-term results of total knee arthroplasty after the use of soft tissue expanders. AB - The success of total knee arthroplasty can be jeopardized by poor wound healing. In the current study, the results of knee arthroplasty after soft tissue expansion were reviewed retrospectively in 27 patients (29 knees) at risk for problematic healing. The incidence of wound complications was recorded for all patients after the expansion procedure and the arthroplasty. A Knee Society score also was calculated at the latest evaluation. Minor wound complications occurred after 21% (six of 29) of the tissue expansion procedures and after 18% (five of 28) of the subsequent arthroplasties. One major wound complication occurred during tissue expansion necessitating abandonment of the planned arthroplasty. No major wound complications occurred in those patients who underwent knee arthroplasty. At an average followup of 34.4 months, the average Knee Society score was 83.7 points. The results of the current study show that the technique of soft tissue expansion before total knee arthroplasty in patients at high risk for wound healing problems can successfully prevent catastrophic wound complications after the arthroplasty procedure and can avoid the need for disfiguring soft tissue reconstructions. PMID- 11064983 TI - Results of all-polyethylene tibial components as a cost-saving technique. AB - Since 1992, the authors have used an all polyethylene tibial component in relatively less active patients older than 70 years of age who present for primary total knee replacement. Results of 312 knee replacements performed between March 1992 and March 1998 are presented in the hopes of showing this technique as a viable cost saving measure. Three hundred twelve primary total knee replacements were performed by the same group of surgeons. Fourteen patients died before the first year postoperative evaluation, leaving an index group of 298 knees in 231 patients. All components were evaluated using the radiographic and clinical parameters of the Knee Society. Hospital for Special Surgery scores also were calculated. All patients were given the Short Form-36 Quality of Life Assessment preoperatively and annually postoperatively. Clinical scores showed dramatic improvement. There have been three revisions (0.7%) but none were for aseptic loosening. Radiographic review revealed 295 replacements with optimum fixation. To date, no patient has osteolysis. The cost differential was significant. The use of this type of tibial component has, to date, afforded excellent clinical and radiographic results. Additional followup is needed to show the longevity of these results and to monitor wear or subsidence of these prostheses. Should these results continue to prove satisfactory in this particular group of patients with relatively low physical demands, significant cost savings may be realized. PMID- 11064984 TI - The role of continuous passive motion after total knee arthroplasty. AB - The usefulness of continuous passive motion after total knee arthroplasty remains controversial. The reported benefits include decreased rates of knee manipulation, deep vein thrombosis, and postoperative use of analgesics, and a greater range of motion. Other studies have reported increased wound complications, bleeding, and pain. Lack of consensus on the use of continuous passive motion exists because reported studies include many confounding variables. Several studies have shown that continuous passive motion in the hospital decreased the rate of knee manipulation from as high as 21% to as low as 0%. Although many studies show that range of motion may improve more rapidly with continuous passive motion, the ultimate range of motion at followup is unchanged. At the author's institution, continuous passive motion is used three times per day (1 hour sessions), beginning on the first postoperative day, within a 4 to 5 day inpatient hospital pathway. Of 132 knees that had a primary posterior stabilized total knee arthroplasty, seven knees (5%) had a manipulation for failure to obtain greater than 70 degrees flexion. No patients had major wound complications that required reoperation. There is no specific charge to the patient for the continuous passive motion because it is included in the hospital per diem charge. The literature and the author's data support the use of continuous passive motion to decrease the rate of manipulation (and its costs) for poor range of motion after total knee arthroplasty. If patients follow fixed inpatient hospital pathways, the length (and possibly cost) of hospital stay is not changed by use of continuous passive motion. The data on the effect of continuous passive motion on overall analgesic use and prevalence of deep vein thrombosis are not clear. PMID- 11064985 TI - Comparison between the kinematics of fixed and rotating bearing knee prostheses. AB - Rotating platform mobile bearing knee implants allow for increased tibiofemoral articular conformity without restricting axial rotation. In the current study, the effect of rotating platform knee replacement with and without posterior cruciate ligament substitution on knee kinematics was investigated. Five knees were implanted sequentially implanted with standard (fixed) bearings and then with rotating platform prostheses, each in posterior cruciate retaining and substituting designs. Three-dimensional kinematics for all knees were measured in an Oxford Knee Rig, which simulates dynamic quadriceps-driven closed kinetic chain knee extension under load. Rotating bearings did not significantly change knee kinematics when compared with fixed bearings. In this in vitro model, the cruciate retaining designs stayed more anterior, and had greater net femoral roll back and tibiofemoral valgus angulation with flexion than cruciate substituting designs. PMID- 11064986 TI - Dome corrective osteotomy for cubitus varus deformity. AB - Between 1994 and 1998, 15 patients had corrective dome-shaped osteotomy of the humerus for posttraumatic cubitus varus deformity. Thirteen patients had surgery before puberty and two patients had surgery after puberty. In the prepuberty group, all the osteotomies were done by a posterior approach with triceps muscle splitting, and cross pins were used to fix the osteotomy. In the postpuberty group, the osteotomies were done by a posterior approach with olecranon osteotomy, and reconstructive plates were used for fixation. The average followup was 2 years and 4 months. Preoperative carrying angle ranged from 19 degrees to 31 degrees varus (average, 26.2 degrees) and postoperative carrying angle ranged from 7 degrees to 15 degrees valgus (average, 10.7 degrees). No loss of correction was observed and all osteotomies united. The preoperative and postoperative differences of the lateral condylar prominence index ranged from 67% to +6% (average, -30.1%). After reviewing these cases, a dome-shaped osteotomy was found to have the following advantages for correction of cubitus varus deformity: the osteotomy site is more stable than a lateral closing wedge osteotomy for maintaining the correction obtained; the domed osteotomy avoids having the lateral condyle becoming prominent; and the posterior scar is more cosmetically acceptable than the lateral scar in the lateral closing wedge osteotomy. PMID- 11064987 TI - Anatomy of the iliolumbar ligament. AB - Information is lacking in the literature on the precise anatomy of the iliolumbar ligament and its individual differences. The morphologic pattern, length, and width of the iliolumbar ligament were determined in 56 embalmed lumbosacral spines from human cadavers. It was possible to classify the iliolumbar ligament into two groups: Type A (74 ligaments), in which anterior and posterior ligaments had separate courses; and Type B (32 ligaments), in which anterior and posterior ligaments moved together as one band. The angle of the posterior iliolumbar ligament in Type A was oriented significantly more posteriorly than that in Type B. The posterior iliolumbar ligament was significantly shorter and oriented more posteriorly in male anatomic specimens than in female ones. PMID- 11064988 TI - Septic arthritis of the hip secondary to rat bite fever: a case report. AB - Rat bite fever is a rare infection typically caused by Streptobacillus moniliformis. The mode of transmission is most commonly through a bite or scratch from an infected rat. This disease is characterized by polyarthritis, fever, and a delayed onset erythematous maculopapular rash of the extremities. The authors report a case of rat bite fever, which led to septic arthritis of the hip. To the authors' knowledge, the complication of hip sepsis requiring an arthrotomy has not been reported in the literature. The orthopaedist should be aware of not only Streptobacillus moniliformis, but also of other zoonotic organisms, which potentially can cause septic arthritis and warrant treatment with specific antibiotics. PMID- 11064989 TI - Arthrofibrosis after total knee arthroplasty. AB - Six total knee arthroplasties in five patients were revised because of persistent limited motion after the primary arthroplasty. All of the revised implants were of an appropriate size and not malpositioned. No cause of stiffness was identified other than soft tissue contracture. Four of the components were posterior cruciate retaining and two were posterior cruciate substituting. Heterotopic bone formation was observed in two knees before the revision surgery and five knees after the revision surgery. Arc of motion was increased from 36 degrees (range, 20 degrees-70 degrees) before revision surgery to 86 degrees (range, 70 degrees-110 degrees) after revision surgery. What triggers the proliferation of extensive scar tissue formation in patients with arthrofibrosis is not clear. Some patients may be predisposed to this condition or may have it develop as a response to the surgical trauma and postoperative rehabilitation. However, when arthrofibrosis does develop after total knee arthroplasty, some improvement in motion and pain can be achieved with revision surgery. PMID- 11064990 TI - Clinical features of multiple epiphyseal dysplasia expressed in the knee. AB - The purpose of this study is to clarify the clinical features of the knee affected by multiple epiphyseal dysplasia. Thirty-one cases of multiple epiphyseal dysplasia were reviewed. Of the patients, 11 were male and 20 were female. The average age at onset of symptoms was 22.5 years. The average age at initial visit to the authors' hospital was 28.9 years. Radiographic findings showed epiphyseal abnormality of the knee in all but two (93%) cases. Irregularity, segmentation of the epiphysis, widening of the joint space, and genu valgum deformity were the dominant findings before epiphyseal closure. After epiphyseal closure, the most characteristic finding was a shallow femoral trochlear groove, which was observed in 56.5% of the cases. Other findings in adult patients included early onset osteoarthritic change, genu valgum, depression of the lateral tibial plateau, and multiple free bodies. However, there still is a possibility that multiple epiphyseal dysplasia exists, even if the patient lacks a shallow femoral trochlear groove. If genu valgum or varum, free bodies, and premature osteoarthritis are observed, one should evaluate other joints, keeping a diagnosis of multiple epiphyseal dysplasia in mind. Patients with knees that have a femoral trochlear groove of normal or near normal shape do exist, and premature osteoarthritic changes may develop in such patients. PMID- 11064991 TI - Granular cell tumors of the extremities. AB - Granular cell tumors are uncommon tumors that may arise from various soft tissue and visceral sites. These lesions often are multifocal but, with rare exceptions, are benign. Much of the literature on granular cell tumors is based on case reports mostly in the otolaryngology literature, and most series are limited to histopathologic studies. There are no documented series of cases in the orthopaedic literature. Granular cell tumors do occur in the extremities; thus, patients with a peripheral granular cell tumor may be referred to an orthopaedist. Ten cases of benign granular cell tumor treated at the authors' institution between 1993 and 1999 are reported. Five tumors of the thigh and one tumor of the groin were treated with wide excision, whereas three tumors of the hand and one tumor of the elbow were treated with marginal resection. Although two tumors treated with attempted wide excision had positive margins, none of the tumors have recurred after an average followup of 27 months. The presentation, radiography, histologic findings, and treatment of benign granular cell tumors that distinguish it from other soft tissue lesions are discussed. PMID- 11064992 TI - Periosteal glomus tumor of the femur: a case report. AB - Osseous abnormalities produced by glomus tumors located in soft tissues of the periungual region have been described. More rare is the location of a glomus tumor within bone, which usually is located in the phalanx of the fingers. However, to the authors' knowledge, there is no previous description of a glomus tumor located in a periosteal location of a long bone. A 50-year-old man with a glomus tumor in a periosteal location of the lower metaphysis of the femur without neoplastic erosion of the cortical surface is reported. Magnetic resonance imaging and intraoperative ultrasonography were needed to locate the lesion. PMID- 11064993 TI - Metastatic lesions of the humerus treated with the isoelastic diaphysis prosthesis. AB - Between January 1, 1987, and December 31, 1997, an isoelastic polyacetal resin prosthesis was used in 50 patients with metastatic bone disease to reconstruct pathologic or impending fractures of the humeral diaphysis. Fifty-seven operations were performed, including seven revision surgeries. The patients were assessed before and after surgery for limb function and quality of life using a modified Karnofsky scale. The mean survival time was 440 days. Ninety-one percent of the operations resulted in restoration or improvement of quality of life. Limb function was good or excellent in more than 80% of the patients after surgery. Breaking of the implant (n = 3), loosening of the implant (n = 2), periprosthetic fracture (n = 1), hematoma (n = 2), infection (n = 1), and one radial nerve paralysis were the main complications. In the cases of implant failure, the prosthesis broke at the site of a locking screw that was inserted across the prosthetic shaft in the cementless implantation technique. This kind of complication could be avoided by using bone cement for implantation or additional plate osteosynthesis between the prosthesis and humeral shaft. The isoelastic diaphyseal prosthesis offers a promising method of treating patients with metastatic lesions of the humeral shaft. PMID- 11064994 TI - Treatment of chronic isolated radial head dislocation in children. AB - Four children had an operative reduction without annular ligament reconstruction for chronic acquired isolated dislocation of the radial head: one girl and three boys with an average age of 7.1 years (range, 4-8.5 years). History indicated that all children had sustained an injury. Results, with an average followup of 50.7 months, were assessed clinically and radiographically. All patients had excellent postoperative functional and radiographic outcomes. Measurements of range of motion of the elbow revealed improved motion in all patients. Reconstruction of the annular ligament seems unnecessary and, therefore, it is suggested that an unreduced radial head dislocation can be treated by simple open reduction and fixation for 6 weeks with a transarticular pin. PMID- 11064995 TI - External fixation for distal radius fractures: effect of distraction on outcome. AB - Few studies have examined the potential adverse effects of excess distraction and prolonged duration of external fixation for the treatment of distal radius fractures. In this study, 19 patients with distal radius fractures treated with external fixation and supplemental Kirschner wire fixation between August 1991 and November 1997 were studied retrospectively. Patients were evaluated by questionnaire, chart review, radiographs, and clinical examination an average of 161 weeks after injury. Although no significant correlation was found between amount of distraction, as measured by carpal height index, and scores for pain, function, radiographs, motion, grip, strength, and final result, a negative correlation was found of all categories with increasing carpal height index. A significant negative correlation was seen between duration of external fixation and scores for pain, motion, and total score, with motion scores being most affected. New York Orthopaedic Hospital grades of good or excellent were attained by 89% of the patients. The data suggest that external fixation with supplemental pin fixation is a satisfactory method of treating severe fractures of the distal radius. Outcome likely is improved with shorter duration of external fixation. PMID- 11064996 TI - Fixation of femoral shaft fractures with a flexible bundle-type nail. AB - In a prospective study, 31 consecutive patients with a femoral shaft fracture were treated with the Marchetti-Vicenzi intramedullary flexible bundle-type nail. Open reduction of the fracture was necessary in 25 of the 31 patients (81%). Twenty-five of the 31 fractures (81%) united within 2.5 to 6 months after the operation (mean, 4.2 months). Nonunion occurred in one patient (3.2%). Other complications included delayed union in five patients (16%), femoral shortening in five (16%), breakage of the distal pins in two (6.5%), and severe varus malunion in two patients (6.5%). Because of the high complication rate in this series, the authors no longer use the Marchetti-Vicenzi flexible nail for treatment of femoral shaft fractures. PMID- 11064997 TI - The anterior T-frame external fixator for high-energy proximal tibial fractures. AB - The authors' experience using anterior T-frame external fixation combined with percutaneous internal fixation for treatment of high-energy proximal tibial fractures is reported. Thirty-six patients (38 fractures) were reviewed who were treated during a consecutive 42-month period. Three patients died and one patient had an amputation for a Type IIIC open injury, leaving 20 males and 12 females with 21 closed and 13 open fractures (two Type II, seven Type IIIA, three Type IIIB, and one Type IIIC). The average followup was 26 months. Fractures united at a mean of 20 weeks. Ten secondary surgical procedures were planned, including seven antibiotic bead removals with autogenous bone grafting and three soft tissue coverage procedures. Nine (26%) complications were found, including one deep infection (septic arthritis) and three pin tract infections, and one each malunion, nonunion, refracture, knee stiffness requiring manipulation under anesthesia, and deep venous thrombosis. The average Knee Society score was 85 for pain and 83 for function. All patients achieved full knee extension and mean flexion was 125 degrees. The anterior T-frame external fixator with percutaneous internal fixation is a reliable method to stabilize these injuries. It is simple, inexpensive, and effective. PMID- 11064998 TI - Mechanical comparison of fixation techniques for the tibial tubercle osteotomy. AB - Tibial tubercle osteotomies currently are used as an exposure technique for revision total knee arthroplasty and for distal patellofemoral realignment. A review of the literature reveals no biomechanical studies that evaluate methods of osteotomy fixation in terms of static strength. This study evaluates the fixation strength of common techniques used to repair tibial tubercle osteotomies. Bevel and stepcut tibial tubercle osteotomies were created in 36 anatomic specimen knees and were repaired with either two 4.5-mm cortical screws or 18-gauge stainless steel cerclage wire. The failure load for the bevelcut osteotomies repaired with two-screws was 1,654 +/- 359 N; for the bevelcut osteotomies repaired with three cerclage wires, 622 +/- 283 N; for the stepcut osteotomies repaired with three cerclage wires, was 984 +/- 441 N; and for the stepcut osteotomy repaired with four cerclage wires, 1,099 +/- 632 N. This study shows that two bicortical screws provide the greatest static fixation strength for repairing tibial tubercle osteotomies. When repairing tibial tubercle osteotomies for distal patellofemoral realignment, screw fixation would provide the most reliable fixation. However, the placement of screws around the stem of a revision arthroplasty tibial component is difficult. Cerclage wires are easier to place and provide solid static fixation, especially with the addition of a proximal stepcut osteotomy. PMID- 11064999 TI - Biodegradable alginate antibiotic beads. AB - The authors investigated the poly-L-lysine-coated alginate beads as an antibiotic delivery system for the treatment of various surgical infections. The sodium alginate was mixed with vancomycin, coated with poly-L-lysine, and lyophilized to form five types of the biodegradable antibiotic beads. Type I, 2.5% alginate, nonpoly-L-lysine coated and nonlyophilized; Type II, 2.5% alginate, poly-L-lysine coated but nonlyophilized; Type III, 2.5% alginate, poly-L-lysine coated and lyophilized; Type IV, 5% alginate, poly-L-lysine coated and lyophilized; and Type V, 7.5% alginate, poly-L-lysine coated and lyophilized. Cytotoxicity of the alginate beads to fibroblasts and HeLa cells was evaluated by the MTT [3-(4,5 dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide] colorimetric assay. A study of in vitro elution of vancomycin of the alginate antibiotic beads was performed. The results suggested that the alginate antibiotic beads present no obvious toxic risk to their use as a drug delivery system. The concentration of vancomycin in these five types of beads was well above the breakpoint sensitivity concentration (the antibiotic concentration at the transition point between bacterial killing and resistance to the antibiotic) for 9,11,12, 14, and 17 days respectively. The release was most marked during the first 3 days. The duration of antibiotic release was prolonged by using techniques of poly-L-lysine coating, lyophilization, and by increasing the content of alginate. This study offers a biodegradable delivery system of antibiotics to treat various surgical infections. PMID- 11065000 TI - Evaluation of strength of healing fractures with dual energy Xray absorptiometry. AB - Dual energy xray absorptiometry was investigated as a method for evaluation of the strength of closed tibial fractures. In 40 goats, a closed midshaft fracture was created in the left tibia. The fractures were stabilized with an external fixator. After 2 weeks (n = 21) and after 4 weeks (n = 19), both tibias were explanted and, using dual energy xray absorptiometry, bone mineral density and bone mineral content were measured in a 1 cm region. With nondestructive bending tests, area ratio and stiffness index were determined and torsional strength and torsional stiffness were determined with a torsional test to failure. Linear regression analysis was used to calculate the squared correlation coefficients for the relations between dual energy xray absorptiometry and the outcome of the mechanical tests. The squared correlation coefficients for the relation between bone mineral density and torsional strength, torsional stiffness, and area ratio and stiffness index were 0.72, 0.76, 0.64, and 0.72, respectively. The squared correlation coefficients for the relation between bone mineral content and these mechanical parameters were 0.72, 0.77, 0.63, and 0.77, respectively. The results using dual energy xray absorptiometry indicate the strength of healing closed fractures. Additional research is required to investigate specific aspects of this technique. PMID- 11065001 TI - The importance of procedure specific training in harvesting periosteum for chondrogenesis. AB - This study was performed to determine the influence of procedure specific and nonspecific training on the chondrogenic potential of explanted periosteum. Seven operators, with varying degrees of orthopaedic surgical experience and procedure specific training in periosteal harvesting, harvested 10 to 16 periosteal explants each from the proximal medial tibiae of 42 New Zealand White rabbits that were 2 months of age. The chondrogenic index assay involved culturing the explants in agarose suspension for 6 weeks, followed by computerized histomorphometric analysis. Chondrogenic indices (the average percent area of cartilage grown in the cultured explants) ranged from 12% to 81% and were influenced strongly by each operator's experience with the technique of periosteal harvesting. Average cartilage yields before practice were in the range of 12% +/- 4% for a technician and 44% +/- 6% for a surgeon, compared with 54% +/ 7% and 79% +/- 2%, respectively, after practice involving more than 300 explants each. Procedure specific experience (with the technique of periosteal harvesting) was more important than the academic qualifications or years of surgical experience in general. These data must be considered when planning or interpreting the results of studies involving periosteal explantation or grafting, or when periosteum serves as a source of mesenchymal stem cells. PMID- 11065002 TI - Growing mass on the medial aspect of the right proximal leg. PMID- 11065003 TI - Changing dietary patterns and cancer prevention: alpha-linolenic acid health risks and benefits. PMID- 11065004 TI - Dietary fat intake and prostate cancer risk: a case-control study in Spain. AB - OBJECTIVES: Epidemiological evidence suggests that dietary factors can play a role in the etiology of prostate cancer. Results from several case-control and cohort studies on nutrient intake and prostate cancer have been unclear. The authors examined the effect of lipid intake on the risk of prostate cancer. METHODS: In order to assess associations between lipid intake and prostate cancer risk, a case-control study was conducted between May 1994 and March 1998 in the Barcelona metropolitan area, Spain. Two hundred seventeen incident cases with histologically confirmed diagnosis of prostate cancer were matched to 434 hospital and community controls by age and residence. Information about food intake was gathered by a semiquantitative food-frequency questionnaire. Unconditional logistic regression was used for the analysis. RESULTS AND CONCLUSIONS: Animal fat intake was associated with prostate cancer with an estimated OR for highest quartile of 2.0 (95% CI 1.2-3.2). Vitamin C intake was inversely associated with prostate cancer (OR = 0.6; 95% CI 0.3-0.9). The prostate cancer risk increased in proportion to alpha-linolenic acid intake. In the analysis adjusting for energy and major covariables the estimated OR for upper quartile of alpha-linolenic acid was 3.1 (95% CI 1.1-3.8). In conclusion, the association between fat intake and prostate cancer may be correlated with alpha-linolenic acid, although the specific mechanism has to be determined. PMID- 11065005 TI - Breastfeeding and breast cancer risk by age 50 among women in Germany. AB - BACKGROUND: Epidemiological evidence which suggests that prolonged breastfeeding protects against breast cancer has accumulated in recent years. Issues with regard to the timing of breastfeeding and effect modification by correlates of breastfeeding and other risk factors of breast cancer remain unresolved. METHODS: A population-based case control family study of breast cancer among women diagnosed by the age of 50, conducted in two geographic areas in Germany, was used to evaluate the effect of breastfeeding on risk of breast cancer. RESULTS: Among parous women in this study (553 cases, 1094 age-matched population controls), having ever breastfed a child for at least 1 month did not confer protection (odds ratio of 0.9 and 95% confidence interval (CI) 0.8-1.2). However, risk of breast cancer significantly decreased with increasing duration of breastfeeding (p for trend = 0.01) and the estimated relative risk was 0.6 (95% CI 0.4-0.9) for 13-24 months of cumulative breastfeeding and 0.5 (95% CI 0.3-1.1) for 25 months or more. Risk was less related to number of children breastfed than to increasing average length of breastfeeding per child (p for trend = 0.03). CONCLUSIONS: The reduction in risk associated with duration of breastfeeding was not primarily due to breastfeeding the firstborn and more evident in women who were older (> 25 years) when they first breastfed and among women who experienced a recent full-term pregnancy. Risks were modified somewhat by a first-degree family history of breast cancer whereby a greater reduction in risk per additional month of breastfeeding was observed among women with a family history than those without (0.9 vs. 1.0). The study results support a protective role of prolonged breastfeeding against the development of breast cancer in predominantly premenopausal women in Germany. PMID- 11065006 TI - Mammography and breast cancer detection by race and Hispanic ethnicity: results from a national program (United States). AB - OBJECTIVE: Some of the racial and ethnic variation in breast cancer incidence rates may reflect differential use of mammography. We report breast cancer rates using mammography and diagnostic data from five race/ethnicity groups. METHODS: Mammography data were analyzed for 573,751 women who received breast cancer screening between July 1991 and March 1998 from the National Breast and Cervical Cancer Early Detection Program (NBCCEDP). Abnormal mammography rates, breast cancer detection rates, and cancer stage distribution data are presented by race/ethnicity and screening round (first or subsequent). RESULTS: For the first screening round, percentages of abnormal mammographies ranged from 7.3% among black women to 9.3% among Asian/Pacific Islander women. Cancer detection rates ranged from 4.9 cancers per 1000 mammograms for Hispanic and American Indian/Alaska Native (AI/AN) women to 7.7 per 1000 for white women. Subsequent round rates were lower but varied similarly. AI/AN women had the highest percentage (68%) of first-round cancers detected in the early stage (range for the other groups: 52-63%). CONCLUSIONS: Breast cancer detection rates for racial and ethnic groups in this program varied less than published population-based incidence rates. Differential use of mammography among these groups may account for some of the variation reported for breast cancer incidence. PMID- 11065007 TI - Case-control study of diabetes, obesity, physical activity and risk of endometrial cancer among Mexican women. AB - OBJECTIVES: Obesity, hypertension and diabetes are closely associated with endometrial cancer (EC). This study evaluates the relationship between diabetes and risk of EC on the basis of obesity. METHODS: A case control study was carried out in Mexico City from 1995 to 1997. Eighty-five histologically confirmed cases were compared with 668 population-based controls obtained through frequency matching. Diabetes status, weight, height and other factors were determined through personal interviews among both cases and controls. RESULTS: Compared to women without diabetes, those with diabetes had an adjusted odds ratio of 3.6 (95% CI = 1.7, 7.4) for EC. This association was modified by body mass index (p interaction < 0.001). Compared to non-overweight and non-diabetic women, non overweight (OR = 3.9. 95% CI = 0.88, 18.0) and overweight (OR = 5.9, 95% CI = 1.6, 21.1) diabetic women had a non-significant elevated risk of EC. However, elevated risk estimates were observed for obese diabetic women (OR = 8.0, 95% CI = 2.8, 22.7). CONCLUSIONS: Our results strongly suggest an interaction effect between obesity and diabetes that significantly increases the risk of EC. This, in turn, may explain the growing number of new EC cases recently observed in developing countries with reduced birth rates and an increased incidence of both obesity and diabetes mellitus. PMID- 11065008 TI - Food group intake and the risk of oral epithelial dysplasia in a United States population. AB - OBJECTIVES: Oral epithelial dysplasia (OED) is a histopathologic diagnosis associated with an increased risk of oral cancer. The paper explores the relationship between OED risk and food group intake. METHODS: In this case control study, incident cases of OED were identified through two oral pathology laboratories. Controls, pair-matched 1:1 to cases on age (+/- 5 years), gender, appointment date (+/- 1 year), and surgeon, were identified through the office in which the respective case was biopsied. Exposure data were obtained via a telephone interview and mailed food-frequency questionnaire. Conditional logistic regression was used to obtain odds ratio point estimates. RESULTS: Based upon 87 matched pairs and after controlling for smoking, drinking, and other potential covariates there was an apparent inverse relationship between OED risk and the consumption of fruits and vegetables, with the intake of these foods being associated with a strong attenuating effect among smokers. OED risk decreased with increased poultry consumption, but increased modestly with bread/cereal and dairy food intake. CONCLUSIONS: This investigation provides evidence that some aspects of diet may be associated with the risk of OED. It also suggests that in oral carcinogenesis the role of diet is not simply one of a late effect. PMID- 11065009 TI - Markers of insulin resistance and sex steroid hormone activity in relation to breast cancer risk: a prospective analysis of abdominal adiposity, sebum production, and hirsutism (Italy). AB - OBJECTIVE: Insulin resistance and increased levels of serum steroids have been hypothesized to be relevant etiological factors for breast cancer. Measurements of markers of insulin resistance and elevated serum steroids may identify women at high risk for breast cancer. The present study analyzed the association of breast cancer with markers of insulin resistance and elevated serum sex steroids, abdominal adiposity, increase in sebum production and hirsutism in a case-control study nested in a prospective cohort study. METHODS: Between 1987 and 1992, 10,786 women (aged 35-69) were recruited in a prospective study on breast cancer in Italy, the ORDET study. Women with a history of cancer and on hormone therapy were excluded at baseline. At recruitment, abdominal adiposity was calculated from the ratio of waist-to-hip circumferences. Sebum production was measured on the forehead under standardized conditions using a sebumeter. Nine androgen sensitive body areas were evaluated for hirsutism and a total hirsutism score was computed. After an average of 5.5 years of follow-up, 144 breast cancer cases were identified among the participants of the cohort. For each breast cancer case, four matched controls were randomly chosen from members of the cohort who did not develop breast cancer during the follow-up period. RESULTS: Waist-to-hip ratio was associated with breast cancer in premenopausal women: age and body mass index (BMI) adjusted relative risk (RR) for the highest tertile of waist-to-hip ratio was 2.2 [95% confidence interval (CI) 1.04-4.75], p for trend 0.03. In the analysis conducted within strata of BMI, the effect of waist-to-hip ratio was confined to the group of thinner women: RR for the highest tertile of waist-to hip ratio was 3.4 (95% CI 1.2-9.5). Sebum production and hirsutism were associated with breast cancer among postmenopausal women. Age and BMI adjusted RRs for the upper tertiles were 2.2 (95% CI 1.1-4.6), p for trend 0.01, and 2.3 (95% CI 1.1-4.9), p for trend 0.03, for sebum and hirsutism, respectively. CONCLUSION: These results add evidence for a role of hormones and metabolic alterations in breast cancer etiology and for different relations of these risk factors with breast cancer in premenopausal and postmenopausal women. PMID- 11065010 TI - Comparison of heterocyclic amine levels in home-cooked meats with exposure indicators (United States). AB - OBJECTIVE: To compare indicators of heterocyclic amine (HCA) exposure with HCA concentrations in home-cooked meat samples. METHODS: Pan-fried hamburger and steak samples were obtained from individuals stating a preference for medium, well done and very well done meat. Concentrations of DiMelQx, IFP, MeIQx and PhIP were determined by HPLC. RESULTS: HCA concentrations at the three doneness levels were not significantly different using the participants' self-reported doneness preference to categorize samples. Using doneness levels determined at the time the meat was cooked and photograph analysis to categorize samples, HCA concentrations increased with doneness level and significant differences were observed between the very well done and lower doneness levels. When assigned to doneness levels by photograph analysis, mean concentrations (ng/g cooked meat) of DiMelQx, IFP, MelQx, and PhIP were 0.18, 0.16, 0.65 and 0.47 in well done hamburger and 0.61, 0.74, 1.88 and 2.04 in very well done hamburger. In steak, mean concentrations were 0.24, 0.10, 0.79 and 0.59 in well done steak and 0.45, 0.14, 1.87 and 0.62 in very well done steak. CONCLUSIONS: HCA levels in home cooked meat samples were significantly different when samples were visually classified for doneness, but not when self-reported doneness preference was used to classify doneness. PMID- 11065011 TI - Skin characteristics and risk of superficial spreading and nodular melanoma (United States). AB - OBJECTIVE: To assess the risk for melanoma associated with moles and pigmentary characteristics. METHODS: Representative melanoma cases (773) among non-Hispanic white residents under age 65 occurring between 1 June 1978 and I December 1983 in Los Angeles County were compared to controls (752) matched to cases by age, sex, race and neighborhood of residence. Factors considered include hair, eye, and skin color; numbers of freckles and moles; and propensity to burn and tan obtained during an in-person interview. RESULTS: Five hundred and fifty-one cases were classified as superficial spreading melanoma (SSM) and 110 as nodular melanoma (NM). For SSM, the important risk determinants were hair and skin color, freckling, and mole prevalence. Light skin and more freckles were found to be more highly associated with SSM for younger compared to older subjects, whereas the associations between SSM and both hair color and moles remained independent of age. NM showed patterns of risk similar to SSM with the exception of skin color. NM showed no evidence of increasing risk with lighter skin, as compared to the strong association seen for SSM. CONCLUSION: Hair and skin color, freckling and, especially, numbers and size of moles are important determinants of melanoma risk. PMID- 11065012 TI - Major depression and cancer: the 13-year follow-up of the Baltimore epidemiologic catchment area sample (United States). AB - OBJECTIVE: The relationship between depression and development of cancer is not well understood, with some studies finding a significant but small increase in risk for cancer among persons with depression. No studies have employed standardized interviews keyed to the diagnostic criteria for Major Depression. Our objective was to evaluate the relationship between Major Depression at baseline and new onset of cancer at follow-up. METHOD: The study was based on a population-based 13-year follow-up survey of community-dwelling adults living in East Baltimore in 1981. After excluding 372 persons with a history of cancer or those whom reported their health as poor at the baseline interview, 3109 adults remained. Information on baseline depression status and cancer at follow-up was available for 2017 persons. A diagnosis of cancer was ascertained at follow-up through interview of survivors and from death certificates. RESULTS: There were 203 new cases of cancer among 2017 persons at risk. Neither Major Depression (relative risk (RR) = 1.0, 95% confidence interval (CI) 0.5-2.1) nor dysphoric episode (RR = 1.3, 95% CI 0.9-1.9) were significantly associated with increased risk of cancer at follow-up. However, among women with Major Depression, the risk of breast cancer was increased (adjusted RR = 3.8, 95% CI 1.0-14.2). CONCLUSIONS: We found no overall association of depression with cancer. However, among women, Major Depression (but not dysphoric episode alone) was associated with the onset of breast cancer. PMID- 11065013 TI - Lifetime occupational physical activity and incidental prostate cancer (Canada). AB - OBJECTIVES: The objective of this study was to assess the risk of incidental prostate cancer associated with occupational physical activity in a population of patients treated for benign prostatic hyperplasia (BPH) by transurethral resection of the prostate (TURP). METHODS: This case-control study was conducted in men aged 45 and over referred for TURP to relieve the symptoms of BPH in one of the eight hospitals of the Quebec City area between October 1990 and December 1992. Cases (n = 64) were all men incidentally diagnosed with prostate cancer and controls were the 546 patients with solely a histological diagnosis of BPH. At the time of their interview, the patients completed a diet history questionnaire and a general questionnaire including a lifetime occupational history. Physical activity was estimated for each job according to data from the US Department of Labor. Logistic regression was used to estimate odds ratios (OR) and 95% confidence intervals (95% CI) of incidental prostate cancer associated with occupational physical activity while adjusting for confounders. RESULTS: A positive association was observed between "ever having a job with sedentary/light work" and incidental prostate cancer (OR = 1.9; 95% CI = 1.1-3.3). ORs for prostate cancer associated with 0%, 1-49%, and > or =50% of life spent in jobs with sedentary/light work were 1.0, 1.6 (95% CI = 0.8-3.1), and 2.5 (95% CI = 1.2 5.2), respectively (p-value for trend = 0.01). Occupational physical activity in the job held during the longest period was inversely associated with prostate cancer: ORs were 1.0, 0.5 (95% CI = 0.2-1.2), 0.4 (95% CI = 0.2-0.9) and 0.2 (95% CI = 0.1-0.7) for sedentary, light, moderate, and high/very high levels, respectively (p-value for trend = 0.008). CONCLUSIONS: The results of this study suggest that physical activity at work could have a beneficial effect on the occurrence of prostate cancer. PMID- 11065014 TI - Endogenous risk factors for childhood leukemia in relation to the IGF system (Greece). The Childhood Haematologists-Oncologists Group. AB - OBJECTIVE: Insulin-like growth factor-1 (IGF-1) and its principal binding protein 3 (IGFBP-3) are central in the mediation of the effect of growth hormone, and the IGF system has been reported to play a role in the pathogenesis of childhood leukemia. METHODS: To further evaluate the hypothesis connecting the IGF system to this disease, we have examined whether IGF-1 and IGFBP-3 are associated with the two main endogenous risk factors for childhood leukemia, namely gender and birth weight, since boys and heavier newborns are known to be at higher risk. IGF 1 and IGFBP-3 were measured under code in the serum of 118 apparently healthy children aged 0-14 years and the values of each of these components were regressed on age, gender and birth weight. Insulin-like growth factor-2 (IGF-2), as a dependent variable, and anemia during the corresponding pregnancy, as a predictor variable, were also evaluated for exploratory purposes. RESULTS: In the total data set, IGF-1 was positively associated with birth weight (p = 0.0001), whereas girls had higher levels of IGFBP-3 (p = 0.01). CONCLUSIONS: It appears that the associations of measured components of the IGF system with the examined risk factors for childhood leukemia are largely compatible with those that would have been expected, if this system played a role in the pathogenesis of childhood leukemia. PMID- 11065015 TI - Smoke-free public areas: a breath of fresh air. PMID- 11065016 TI - An appraisal of the literature on centric relation. Part I. AB - The literature directly and indirectly related to centric relation (CR) has been reviewed chronologically. More than 300 papers and quoted sections of books have been divided into three sections. The first two parts are related to CR. Studies in this group mainly compared either the position of the mandibular condyle or the mandible itself in different CR recordings. Various tools were discussed for this purpose. The third part of the paper is about CR-centric occlusion (CO) discrepancy. CR still remains one of the controversial issues in prosthodontics and orthodontics. Debates such as mounting casts on the articulator by reproducible records for orthodontic treatment planning and end results, and whether or not orthodontic treatment based on CO causes temporomandibular joint dysfunction, remain unsolved. The references are listed at the end of Part III. PMID- 11065017 TI - Amitriptyline treatment of chronic pain in patients with temporomandibular disorders. AB - Randomized clinical trials of amitriptyline will require data from pilot studies to be used for sample size estimates, but such data are lacking. This study investigated the 6-week and 1-year effectiveness of low dose amitriptyline (10-30 mg) for the treatment of patients with chronic temporomandibular disorder (TMD) pain. Based on clinical examination, patients were divided into two groups: myofascial and mixed (myofascial and temporomandibular joint disorders). Baseline pain was assessed by a Visual Analogue Scale (VAS) for pain intensity and by the McGill Pain Questionnaire (MPQ). Depression was assessed by the Beck Depression Inventory (BDI) short form. Patient assessment of global treatment effectiveness was obtained after 6 weeks and 1 year of treatment by using a five-point ordinal scale: (1) worse, (2) unchanged, (3) minimally improved, (4) moderately improved, (5) markedly improved. The results showed a significant reduction for all pain scores after 6 weeks and 1 year post-treatment. The depression scores changed in depressed but not in non-depressed patients. Global treatment effectiveness showed significant improvement 6 weeks and 1 year post-treatment. However, pain and global treatment effectiveness were less improved at 1 year than at 6 weeks. PMID- 11065018 TI - Biomechanical effects of double or wide implants for single molar replacement in the posterior mandibular region. AB - Double implants have been thought to have biomechanical advantages for single molar replacement. To evaluate the effectiveness of double implants versus a wide implant, the vertical forces and torque on each implant were calculated by three dimensional geometric analysis. Buccal load (100N) perpendicular to cuspal inclination (20 degrees) was applied at the occlusal surface of the superstructure. The three kinds of load points (A, B, C) were 1.5, 3.5, and 5.5 mm from the mesial contact point, respectively. Three implants were compared: mesial and distal double implants (phi 3.3 mm), and a wide implant (phi 5 mm). The wide implant showed torque around the long axis (1.8-15.0 N x cm) whereas double implants had no torque. On the other hand, the vertical forces on the mesial double implant were both smaller (60%: loaded at point C) and larger (140%: loaded at point A) than the wide implant. Given the smaller surface area of the mesial double implant, this large force may generate much higher stress in the peri-implant bone. These results suggest that the biomechanical advantage of double implants for single molar replacement is questionable when the occlusal force is loaded at the occlusal surface near the contact point. PMID- 11065019 TI - Clinical evaluation of an urethane tetramethacrylate-based composite material as a prosthetic veneering agent. AB - The purpose of the current study was to evaluate the clinical performance of a hybrid composite material used as a prosthetic veneering agent after servicing for more than 4 years. A photo-curable composite material (Cesead) was selected as the veneering agent. Composite resin veneered restorations made with the composite and a noble metal alloy were inserted into 110 teeth of 40 patients. Modified United States Public Health Service (USPHS) criteria were used for direct evaluation of colour matching ability, veneer-metal interfacial staining, veneer surface texture, staining of the metal casting, and wear of the veneer metal interface. After an average observation period of 5 years and 3 months, the rate of restorations considered to be clinically ideal (Alfa) was 75.5% for colour match, 93.6% for veneer-metal interfacial staining, 82.7% for veneer surface texture, 97.3% for staining of the metal casting, and 95.5% for wear. The restorations judged as clinically unacceptable (Charlie) consisted of only one case for colour match and three cases for veneer surface texture. Although the Alfa rate decreased with increasing length of service, the Cesead composite is considered to be a clinically reliable material as a prosthetic veneering agent. PMID- 11065020 TI - Fitting a temporomandibular joint prosthesis to the skull. AB - Fitting a temporomandibular joint (TMJ) prosthesis to the skull by using stock prostheses seems to be an appropriate method. However, fitting the skull with one stock part requires many differently shaped parts. Therefore, we fitted the skull with two connected stock parts. The aim of the study was to test whether it is possible to achieve a close fit to the skull with this design, with a maximum of 10 different parts. The articular eminence was fitted with a gully-shaped fitting member, which was rotationally connected to a basic part that fitted to the lateral side of the TMJ. The relevant dimensions of 20 dry skulls were measured and the results were used to derive the optimal dimensions of the prosthesis parts. Prototypes were subsequently fabricated. The fit of the prototypes was tested by measuring the maximum gap between fitting member and skull. All skulls could be fit with a set of four different basic parts and three different fitting members. The average maximum gap between fitting member and skull was 0.20 mm (range 0.11-0.43 mm). It was concluded that a close fit to the skull can be achieved with two connected stock parts and with a total number of seven parts. PMID- 11065021 TI - Morphological and positional assessments of TMJ components and lateral pterygoid muscle in relation to symptoms and occlusion of patients with temporomandibular disorders. AB - Disc displacement is accepted as one of major findings in temporomandibular disorders (TMD). However, the associations of disc positions with morphological and positional changes of temporomandibular joint (TMJ) components and lateral pterygoid (LP), TMD clinical symptoms, and occlusion have rarely been discussed quantitatively. In this study, the morphological and positional changes of TMJ components and LP were assessed by means of magnetic resonance imaging (MRI) and tomography of the TMJ in 41 TMD and nine control (CN) subjects. Disc positions in TMD subjects were divided into normal position (NP) and anterior displacement with and without reduction (ADR+ and ADR-, respectively). From MRI scans and tomograms, the morphological and positional changes of TMJ components and LP were measured and compared among CN, NP, ADR+ and ADR- groups. Correlations between these measurements and the scored clinical symptoms and occlusal factors were analysed in TMD subjects. The results indicated that: (1) TMJ osseous structures and LP showed no significant difference among CN and the three TMD groups, except for a posterior seat of condyle and shorter/steeper condylar movement during jaw opening; (2) disc length and inclination were significantly shorter and steeper, respectively, in ADR+ and ADR-; (3) disc positions were not specified by clinical symptoms and occlusal factors, except for the dominant TMJ sounds in ADR+; (4) an uncoordinated movement of the condyle/disc complex was found in ADR+ and/or ADR-; (5) TMJ osseous structures and the disc were weakly associated with clinical symptoms and occlusal factors. However, the LP showed negative associations with palpable pain for both the TMJ and jaw muscles and the static occlusal factors. These findings suggest that TMJ internal derangements are more related to the positional changes or spatial relationships of TMJ components but less to the individual morphologies of TMJ osseous structures, disc and LP, as well as specific clinical symptoms and occlusal factors, which might be in disagreement with a large body of previous statements. PMID- 11065022 TI - Erosion of dentine and enamel in vitro by dietary acids: the effect of temperature, acid character, concentration and exposure time. AB - Sales of soft drinks has been increasing by 56% over the last 10 years and are estimated to keep rising at about 2-3% a year. Further, the reported incidence of tooth erosion has been increasingly documented. Whilst these factors could well be linked, many individuals with erosive diets are not presenting with erosion. This would suggest the effects of many variables, hence the aim of these investigations. Methodologies included preparing enamel and dentine samples from unerupted human third molars. Groups of five specimens were placed in citric acid over a temperature range of 5-60 degrees C for 10-min exposures; placed in citric, lactic, malic or phosphoric acid (0.05, 0.1, 0.5, and 1% (w/v)) for 10 min exposures; and placed in the same three organic hydroxy acids at 0.3% (w/v) or phosphoric acid at 0.1% (w/v) for 3 x 10-min exposures. Tissue loss was determined by profilometry. Results showed that increasing temperature, concentration and exposure time increased the erosion of dentine and enamel. This study has shown that under highly controlled conditions, erosion of dentine and enamel by dietary acids can be greatly influenced in vitro by temperature, concentration, type of acid and exposure time. These factors could be employed in order to reduce the erosivity of soft acidic drinks. PMID- 11065023 TI - Relationship between oral function and occlusal support in denture wearers. AB - Patients with removable partial dentures or complete dentures do not demonstrate masticatory function to the same level as patients with a full set of natural teeth. The purpose of this study was to characterize the relationship between reduction of masticatory function, in terms of masticatory performance and bite force, and the existence of remaining natural occlusal supports as assessed by the Eichner index. One hundred and eighteen removable partial denture and complete denture wearers were selected for analyses. These subjects were divided into four groups depending upon the number of occlusal supports. Seventy dentate subjects with full occlusal support were designated as a comparison group. Bilateral bite force was measured at the first molar region in all subjects. Masticatory performance was assessed using the modified Masticatory Performance Index. Peanuts were used as the test food. Both bite force and masticatory performance were significantly associated with group classification. Moreover, both bite force and masticatory performance of the four denture groups were significantly reduced compared to the comparison group and this tendency was remarkable for the denture groups without occlusal support. These results suggest that the existence of functional tooth units may be a key factor in preservation of masticatory function. PMID- 11065024 TI - Functional subdivision of the human masseter and temporalis muscles as shown by the condylar movement response to electrical muscle stimulation. AB - In previous studies from our laboratory, a functional subdivision of the human temporalis and masseter muscles was demonstrated by means of opto-electronic recordings of the lower incisal point movement responses to electrical muscle stimulation. In the present study, it was examined whether this subdivision was also reflected in different movement responses of the mandibular condyle. To that end, the condylar movement responses to unilateral stimulation of four masseter muscle parts and three temporalis muscle parts were studied in four different jaw positions. The kinematic centre was used for condylar reference point. For both the amplitude and the direction of the movement responses, the effects of stimulation location and jaw position were studied using multivariate ANOVA and contrast analyses. It was found that for both outcome variables, the functional subdivision of the masseter and temporalis muscles was also reflected in some, but not all, of the movement responses of the mandibular condyles. The deep masseter muscle part and the (anterior) temporalis muscle part responded similarly to electrical stimulation. PMID- 11065025 TI - Morphological differences in individuals with lip competence and incompetence based on electromyographic diagnosis. AB - The study group consisted of 19 subjects with positive overjet and overbite, and 17 subjects with skeletal open bite. Two bipolar surface electrodes were attached to the skin of the upper and lower lips. The mean integrated amplitude of the electromyographic (EMG) activity was obtained at the mandibular rest position with the lips in contact and with the lips apart. Subjects were divided into two groups based on positive or negative values of the difference in integrated EMG activity of the mentalis muscle between the two lip positions. Subjects displaying a negative value were classified as having competent lips and those displaying a positive value were classified as having incompetent lips. The EMG activity of the mentalis muscle was found to be more indicative of lip sealing as compared with the EMG activity of the depressor of the lower and the upper lips. The activities of the mentalis muscle at the mandibular rest position with the lips in contact and with the lips apart appear to offer an objective criterion for the evaluation of lip incompetence. In addition, the vertical dimension of the face, as well as the proclination of the incisors, appear to affect lower lip function. PMID- 11065026 TI - Asymmetry in jaw-jerk reflex latency in craniomandibular dysfunction patients with unilateral masseter pain. AB - The purpose of the present study was to elucidate the effects of unilateral masseter muscle pain on the jaw-jerk reflex. The latency and peak-to-peak amplitude of bilateral electromyographic activity recorded at the masseter muscles during the jaw-jerk reflex were measured in 18 patients with craniomandibular dysfunction (CMD) with strictly unilateral masseter pain or tenderness and 10 control subjects using a computerized recording and analysis system. The reflex was elicited, at the mandibular rest position, by tapping the centre of the chin downwards with a reflex hammer incorporating a microswitch that triggered the sweep of the recording apparatus upon contact with the chin. In the CMD group, the jaw-jerk latency on the affected side (6.89 +/- 0.98 ms) was significantly shorter (P < 0.01) than that on the unaffected side (7.59 +/- 0.92 ms). In the control group, there was no difference between the jaw-jerk latencies on the right (7.06 +/- 0.64 ms) and the left (7.08 + 0.65 ms) sides. The range of side asymmetry for jaw-jerk latencies in the CMD group was greater than that in the control group. In six patients, the latency difference exceeded 1 ms. The asymmetry of latency of the jaw-jerk reflex was thought to be due to facilitation on the side with masseter pain or tenderness. This facilitation on the ipsilateral side might be produced by enhanced gamma drive induced by sustained nociceptive stimulation. Such effects may be related with clinically derived concepts regarding such muscle dysfunction as myospastic activity or trigger points. PMID- 11065027 TI - The relation of canine guidance with laterotrusive movements at the incisal point and the working side condyle. AB - The effect of different types of canine guidance on the patterns of laterotrusive tracing at the incisal point and the relationship between the laterotrusive inclinations and the working side condylar movements were investigated in 42 young subjects. The subjects were divided into M and D groups according to their mesial and distal canine guidance, and were also divided into protrusive laterotrusion (PL) and retrusive laterotrusion (RL) groups according to their laterotrusive tracing patterns. No differences of laterotrusive inclinations and working side condylar movements were found between the M and D groups. The laterotrusive tracing pattern had no corresponding association with the type of canine guidance. In relation to the movements of the working side condyles, significant differences were found between the PL and RL groups. The condyles moved laterally and posteriorly in the RL group, but moved lateral and inferior in the PL one. The distance of condylar movement in the X direction was correlated with the horizontal and sagittal inclinations of laterotrusion. The results indicate that the movements of the working side condyle were affected functionally by the laterotrusion, but not by either the mesial or the distal type of canine guidance. PMID- 11065028 TI - The amalgamation of two colleges--an opportunity to look at the rapidly changing world scene. PMID- 11065029 TI - Anal incontinence in women with third or fourth degree perineal tears and subsequent vaginal deliveries. AB - We contacted 208 women 13 years after they suffered an obstetrical anal sphincter tear in order to estimate the effect of subsequent vaginal deliveries on anal continence. Among the 177 eligible responders, 129 sustained a partial or complete 3rd degree and 48 a 4th degree tear; 114 women had subsequent vaginal deliveries. Anal incontinence was more common in women with 4th (25.0%) than with 3rd degree tears (11.5%, p = 0.049). Subsequent vaginal deliveries were associated with a higher prevalence of severe incontinence in women with 4th degree tears (p = 0.023). No aggravation or increase in prevalence of incontinence was observed in women with 3rd degree tears. These results suggest that in a subsequent pregnancy, careful evaluation is necessary and an abdominal delivery may be advisable for women with previous major sphincter trauma. PMID- 11065030 TI - A survey of pregnant women's attitude towards breech delivery and external cephalic version. AB - A structured interview survey was carried out in 150 women who came for their first antenatal visit in a university hospital in Hong Kong between June and July 1998. Their opinions and perceptions of fetal and maternal safeties on different modes of delivery for both cephalic and breech presentation, and external cephalic version (ECV) were surveyed. Their decisions on the management of term breech-presenting pregnancy were examined. Most women (92%) preferred vaginal delivery to Caesarean delivery (CS) in case of cephalic presentation, mainly because it was a natural way of parturition. They perceived that vaginal delivery was safer than CS for both mothers and babies, but the reverse was true for breech presentation. About 82% chose ECV as the first choice of managing breech presentation, mainly because a successful version allowed a natural way of delivery. Only 2% of women considered ECV ineffective, and 13.3% and 18.7% considered it not safe for mothers and fetuses respectively. Therefore, ECV should be an available option in all obstetric units. Adequate counselling and explanation would improve the acceptance of ECV. PMID- 11065031 TI - Seroprevalence and assessment of risk factors for hepatitis C virus infection in pregnancy. AB - Screening for hepatitis B is routinely performed in most antenatal clinics. Whether the same should occur for hepatitis C needs to be assessed for each population by determining the prevalence of this infection within the community and whether any particular high-risk group can be identified. A series of 2,000 consecutive patients attending for antenatal care at the Mercy Hospital for Women, Melbourne, was tested for evidence of hepatitis C infection. The prevalence of hepatitis C infection in this group was 1.45% (95% confidence interval 0.97-2.1%). Significant independent risk factors were a history of intravenous drug use, blood transfusion and previous pregnancy ending prior to 20 weeks' gestation. Currently no treatment exists for hepatitis C and as there are no effective means of preventing transmission to the baby, routine screening cannot be justified in view of the low prevalence of this infection among antenatal patients. Selective screening of patients with relevant risk factors for hepatitis C should be carried out as the most efficient and cost-effective strategy in pregnancy. PMID- 11065032 TI - A randomised controlled trial comparing birthing centre care with delivery suite care in Adelaide, Australia. AB - Birthing centre care offers women with a low risk of complication in pregnancy an alternative to conventional care for the birthing of their baby. It is important these two forms of care are appropriately assessed. A randomised controlled trial comparing the newly opened birthing centre with the established conventional delivery suite was conducted at the then Queen Victoria Hospital, Adelaide, South Australia. The outcomes measured included maternal satisfaction, costs and clinical outcomes both for mother and baby which related to the need for Caesarean section, episiotomy or tear rate and method of feeding. Two hundred and one women attending the hospital's antenatal clinic were randomly allocated to either birthing centre or delivery suite care. One hundred women were allocated to the birthing centre. No differences were found in either group related to clinical outcomes or costs. The only difference in maternal satisfaction was the choice women made for their next birth. More women in the birthing centre group felt they were encouraged to breastfeed immediately after birth. While the numbers in this study were too small to detect any but large differences in outcome, birthing centre care should remain an option for women and further studies undertaken with larger numbers. PMID- 11065033 TI - Occipital posterior and occipital transverse positions: reappraisal of the obstetric risks. AB - Malpositions in labour in a vertex-presenting fetus are known to be associated with increased risks of operative delivery A retrospective analysis of all deliveries over 4 years in a university teaching obstetric unit was performed using the available obstetric database. All cases of live births with cephalic presenting babies after 36 completed gestational weeks were analysed, and included 17,533 out of 20,533 total deliveries over the study period. The study group included those cases with occipital posterior and transverse positions, based on the documentation of the position of the vertex at the time of delivery, or at the last clinical examination before obstetric intervention, while occipital anterior cases constituted the control group. The overall incidence of malpositions was 14%, and operative delivery rate in the study group was 82.5% versus 20.7% in the control group. After excluding cases of operative delivery for non-mechanical indications, such as fetal distress, the adjusted odds ratio for the malposition group was 9.8 (95% CI 8.91-10.8) for total assisted delivery and 30.2 (95% CI 25.6-35.5) for Caesarean section compared to the occipital anterior group. Malpositions are definitely associated with a marked increase in the risk of operative delivery including Caesarean section. Labour complicated by malposition should be considered high risk, and should warrant due preparation for obstetric intervention. PMID- 11065034 TI - Pregnancy outcome in women with reflux nephropathy and the inheritance of vesico ureteric reflux. AB - In women with reflux nephropathy, we investigated whether pre-existing hypertension and impaired renal function influence the rates of preeclampsia, renal function deterioration and preterm birth. The infants were investigated for vesico-ureteric reflux (VUR). A prospective audit of 54 pregnancies in 46 women with reflux nephropathy was performed. Preeclampsia complicated 24% of pregnancies and was increased in women with pre-existing hypertension (42%) compared with normotensive women (14%), (RR 3.0 (95% CI 1.1-7.8)). Nine (18%) women experienced deterioration in renal function during pregnancy Women with mild or moderate renal impairment were at increased risk of renal function deterioration (RR 12.7 (95% CI 1.6-98.5); RR 19.8 (95% CI 2.6-155)), respectively A third of infants were delivered preterm. The risk of preterm birth was increased if the mother had pre-existing hypertension (p = 0.01) or moderate renal impairment (p = 0.002). Seventeen (43%) of the 40 infants who underwent micturating cystourethrography had VUR, consistent with autosomal dominant inheritance with reduced penetrance. In reflux nephropathy, pre-existing hypertension was associated with an increased risk of preeclampsia and pre existing renal impairment with deterioration in renal function. Infants of women with reflux nephropathy should be screened for VUR. PMID- 11065035 TI - Haematopoietic indicators of fetal metabolic acidosis. AB - We aimed to study the haematopoietic response in normal and acidotic deliveries following vaginal and abdominal delivery and to compare this to the surrogate markers of perinatal acidosis. Blood gas analyses, complete blood pictures and erythropoietin assays were performed on umbilical or early neonatal blood samples. Placental sections were examined for the presence of nucleated red blood cells. Perinatal clinical risk factors and major neonatal outcomes were collected. The control population was 78 deliveries where the cord arterial pH was > 7.10. Controls born after labour were compared to those born prior to the onset of labour and to 14 acidotic infants born after labour. Nucleated red blood cells did not increase with labour in the control groups but were significantly higher (p < 0.05) in the acidotic group. Erythropoietin did not significantly change with either labour or acidosis. The predictive values from nucleated red blood cell counts were higher than those from low Apgar scores, atypical cardiotocograph traces, meconium-stained amniotic fluid, erythropoietin and the presence of nucleated red blood cells in placental sections. Nucleated red blood cell counts may be a useful surrogate marker of acidosis where blood gas analysis is unavailable. Further studies are required to examine the timing of the increase of erythropoiesis to help define the onset of the stimulus. PMID- 11065036 TI - First trimester ultrasound with nuchal translucency measurement for Down syndrome risk estimation using software developed by the Fetal Medicine Foundation, United Kingdom--the first 2000 examinations in Newcastle, New South Wales, Australia. AB - In September 1997 screening for Down syndrome using first trimester ultrasound to measure nuchal translucency, with risk estimation by the software program developed in the United Kingdom by the Fetal Medicine Foundation, was introduced in Newcastle, New South Wales. In the first 2,000 such risk estimations 134 women (6.7 %) were screen positive (with a risk of greater than 1 in 300 at that gestation for Trisomy 21). In the first 1,000 of these 2,000 fetuses delivered thus far there were 8 cases of Trisomy 21, 2 of Trisomy 18 and 1 of 47 XXX. Nine of these 11 were screen positive, the only false negative results being for 2 cases of Trisomy 21. The detection rate for Trisomy 21 was 6 out of 8 (75%) and for every case of Trisomy 21 (Down Syndrome) detected by this process, 11.3 invasive tests would have been needed to make that diagnosis in a screen positive woman. PMID- 11065037 TI - Ultrasonic fetal measurements: new Australian standards for the new millennium. AB - In over 30 years of ultrasound assessment of the fetus, Australian researchers have only produced growth curves for the biparietal diameter (BPD) and occipito frontal diameter (OFD) for general use. The overseas curves used for other fetal parameters are up to 25 years old and based on predominantly white middle class sample populations. In the last decade the ethnicity in Australia has changed significantly, putting into question the accuracy of the existing charts. This 3 year study of 3,800 pregnancies has resulted in the production of fetal measurement charts for the BPD, OFD, head circumference (HC), abdominal circumference (AC), crown rump length (CRL), femur and humerus lengths. Using over 11,600 measurements collected from diverse ethnic, social and economic groups within the Australian population, rigorous statistical analysis was performed. The results showed that statistically significant differences occur between the curves currently in regular use and those for the OFD, HC, AC, CRL and humerus length obtained from our data. PMID- 11065038 TI - The management of pulsion enterocoele with the Zacharin abdominoperineal technique (and mesh sacrocolpopexy). AB - Our aim was to evaluate the technique, results and complications in 126 women with pulsion enterocoele treated with the combined abdominoperineal repair. Mean follow-up was 25 months with a range of 0-83 months. Operative morbidity included bladder trauma (0.7%), bowel injury (3.1%), wound breakdown (0.7%), infection (10%), pulmonary embolus (0.7%) and blood transfusion (40%). Longterm complications included prolonged urinary retention (11%), incisional hernia (4.7%) and constipation (40.4%). 92.4% of women were cured. PMID- 11065039 TI - Sarcomas and the conservative management of uterine fibroids: a cause for concern? AB - The introduction of conservative management options has further increased the choices available to clinicians treating women with symptomatic uterine fibroids. However, in the absence of a tissue diagnosis, the possibility of mismanaging an underlying uterine sarcoma is still present, placing these patients at potential risk of a delayed diagnosis of this serious pathology. Evidence suggests that 1 in 250-400 women presenting with what are thought to be symptomatic fibroids, will in fact have an underlying sarcoma, making this an important clinical issue. This paper therefore reviews the methods currently available for the assessment of women in whom conservative management of symptomatic fibroids is contemplated. PMID- 11065040 TI - Improved patient acceptability with a transdermal drug-in-adhesive oestradiol patch. AB - The aim of this trial was to assess the relative patient acceptability of two transdermal oestradiol patches used in treatment of oestrogen deficiency in postmenopausal women. Thirty-five hysterectomised postmenopausal women with no previous experience of transdermal oestradiol delivery systems received treatment with either once-weekly drug-in-adhesive (DIA) patches or twice-weekly reservoir patches for 4 weeks, and were then switched to the alternative treatment for a further 4 weeks. At the end of the study, the patients completed a questionnaire to assess their relative preference for a number of characteristics of the 2 transdermal systems and, where possible, their preference for transdermal compared with oral hormone replacement therapy. Thirty-one patients completed the study; four withdrew during treatment with the reservoir patch. The DIA patch was preferred for being 'easiest to remember to apply' by 80% of patients (p < 0.01), 'easiest to open' and 'easiest to apply' by 68% (p = 0.025), and as having 'best cosmetic appearance' by 65% (p = 0.05) and 'best overall skin adhesion' by 61% (p < 0.01). While 10% of patients rated the reservoir patch as 'least irritating to the skin' (p = 0.03), only one patient found this patch 'most comfortable to wear' (p < 0.01). The DIA patch was selected by 87% of patients as their preferred treatment overall (p = 0.001). Ninety-one per cent of 22 responding patients were at least as confident of treatment with transdermal patches as with oral hormone replacement therapy (p = 0.006) and 74 % of 27 responders preferred transdermal to oral treatment (p = 0.004). The DIA patch appears to be more acceptable to patients than the reservoir patch as a transdermal oestradiol delivery system for the treatment of postmenopausal oestrogen deficiency. Characteristics of the DIA patch which may account for improved patient acceptance include ease of remembering once-weekly patch application, improved cosmetic appearance and comfort, and better adhesion. PMID- 11065041 TI - Cloning: its relevance to monozygotic twins. PMID- 11065042 TI - A prospective study of gestation and birthweight in Aboriginal pregnancies in far north Queensland. AB - This study aimed to determine the cause(s) of the increased incidence of low birthweight birth in Aboriginal pregnancies. The study prospectively examined a cohort of Aboriginal women presenting for antenatal care before 20 weeks gestation (ultrasound proven) and a reference cohort of Caucasian women in four remote North Queensland communities served by the Far North Regional Obstetric and Gynaecological Service (FROGS) and the antenatal clinic at Cairns Base Hospital. Women with no known medical factors affecting fetal growth or gestation were recruited. Of the 102 Aboriginal and 101 Caucasian women recruited, 96 Aboriginal and 96 Caucasian women completed the study, providing groups of sufficient size to allow statistical assessment at 80% power and 95% significance. Outcomes measured were gestation at delivery, planned or spontaneous birth, neonatal anthropometric measurements and Dubowitz score. The phenotypic and demographic characteristics of the women, their pregnancies, and their babies were also compared to understand the major associations with low birthweight birth in the Aboriginal women. Apart from Aboriginal ethnicity, excessive alcohol use in pregnancy, low maternal body mass index (BMI), and low maternal age had significant negative correlations with birthweight, and excessive tobacco use in pregnancy and high maternal gravidity showed strong similar trends. Culturally appropriate programs need to be funded and developed to reduce the incidence of low birthweight Aboriginal birth, rather than medical programs primarily aimed at the reduction of the incidence of preterm labour. PMID- 11065043 TI - Maternal stress and obstetric and infant outcomes: epidemiological findings and neuroendocrine mechanisms. AB - This review examines the associations between antenatal maternal stress and obstetric and infant outcomes using preterm delivery as the key outcome indicator. This was done by means of a Medline search focusing predominantly on prospective, controlled studies which investigated both the associated epidemiological factors and putative neuroendocrine mechanisms. There is evidence from a number of United States studies in economically deprived African American women for an association between perceived maternal life event (LE) stress and preterm delivery. The findings from the European studies are conflicting, partly because they combine outcome measures ie. preterm delivery and low birth weight. However the three largest Scandinavian epidemiological studies examining preterm delivery and controlling for confounders such as smoking, age and obstetric history, have confirmed this association. These studies taken together suggest that this may be a robust finding not limited to socioeconomically deprived African American samples and independent of other significant risk factors. Two small prospective studies examining the relationship between the hypothalamic pituitary-adrenal (HPA) axis, psychosocial status and premature delivery have reported a significant association between a set of adverse psychosocial factors on the one hand, and levels of adrenocorticotrophic hormone (ACTH), corticotrophin releasing hormone (CRH) and cortisol levels, and on the other hand, a significant correlation between CRH levels and premature delivery. Clearly, these findings remain preliminary and indicate a complex relationship between perceived stress in pregnancy, the HPA axis and premature delivery. The impact of antenatal maternal stress on infant temperament and psychopathology remains to be examined more fully in prospective controlled trials. PMID- 11065044 TI - Universal antenatal group B streptococcus screening? The opinions of obstetricians and neonatologists within Australia. AB - Group B streptococcus (GBS) is the leading infectious cause of morbidity and mortality in Australian newborns. Although intrapartum chemoprophylaxis is recommended to reduce the risk of neonatal GBS transmission and disease, controversy exists as to the best method to select women 'at risk' for this treatment. Our study aimed to survey the opinions of obstetricians and neonatologists currently in practice in Australia on GBS screening and treatment. Of the 488 obstetricians and 68 neonatologists currently in practice who responded to the survey, 271 obstetricians (56%) and 40 neonatologists (61%) supported universal antenatal screening. Of those respondents who did not support a universal antenatal screening policy, 196 (93%) and 24 (92%) of the obstetricians and neonatologists respectively, supported antenatal screening based on risk factors. This diversity in practitioner opinion highlights the lack of certainty in the literature as to the best management strategy to prevent neonatal GBS sepsis. PMID- 11065045 TI - Early results of chemo-radiotherapy for the definitive treatment of cervix cancer. PMID- 11065046 TI - Why is the placenta being ignored? AB - The relationship between the frequency of published recommended indications for placental pathological examination and the frequency of requests for such examination in a population-based study of term newborn encephalopathy was examined. Only 11.2% of placentas among 276 case infants and 0.7% of placentas among 564 term control infants were examined. Using the criteria set out in a consensus statement by the American College of Pathologists, all 276 cases fulfilled multiple maternal, fetal and placental indications for placental examination. Furthermore 43.3% of control infants fulfilled at least one criterion. Of the 25 case placentas that underwent pathological review, 16 were reported as having no diagnostic abnormality Six cases (24%) showed clinically important findings: four had evidence of infection, one had multiple chorangiomata and one had thrombosis and rupture of the umbilical vein. Of the three remaining placentas, one showed funisitis, one showed minor lymphohistiocytic villitis and one was from monochorionic twins. To our knowledge there are no agreed Australian guidelines for when a placenta should be submitted for pathological examination. We suggest that until guidelines based on properly designed studies are developed it may be appropriate to store all placentas for at least 72 hours. If the infant develops neurological symptoms or requires unexpected admission to a neonatal intensive care unit then placental examination may reveal important aetiological diagnostic and prognostic information. PMID- 11065047 TI - Cervical pregnancy treated with a single intravenous administration of methotrexate plus oral folinic acid. AB - Cervical pregnancy is a rare form of ectopic pregnancy and at present there is no consensus as to the most appropriate treatment. A patient with a 6-week cervical pregnancy was successfully treated with a single intravenous administration of methotrexate and oral folinic acid without any adjuvant therapy Her recovery was uncomplicated. She subsequently conceived and was delivered by Caesarean section of a live healthy male baby. The efficacy of the described regimen in the management of a cervical pregnancy requires further assessment. This modality of treatment does not appear to adversely affect fertility and future pregnancies. PMID- 11065048 TI - Spontaneous rupture of pregnancy in a hemicorpus of complete bicornuate uterus. PMID- 11065049 TI - An unusual case of a retroperitoneal mass causing urinary retention in a 40-year old woman. PMID- 11065050 TI - Menorrhagia caused by dengue fever. PMID- 11065051 TI - Giant urachal cyst in utero. PMID- 11065052 TI - Malignant lymphoma of uterus: a case report with a review of the literature. AB - The female genital tract is rarely the initial manifestation site of malignant lymphomas. Most genital lymphomas arise in the vagina or cervix while those of the uterine corpus are extremely rare. Patients usually present with bleeding, abdominal or pelvic discomfort or back pain but, very infrequently, the tumours are discovered as a result of a routine examination. Our patient was a 67-year old postmenopausal woman presenting with haematuria and upper abdominal pain. She had several investigations for haematuria including cystoscopy, intravenous urography (IVU) and both renal and pelvic scans. The pelvic scan revealed an enlarged uterus with some calcification suggestive of a fibroid uterus. An abdominal hysterectomy was performed. Histopathology revealed non-Hodgkin's malignant lymphoma of the uterine corpus. She subsequently had post-operative chemotherapy. PMID- 11065053 TI - The detection, investigation and management of hypertension in pregnancy. PMID- 11065054 TI - Estimation of fetal weight by ultrasound. PMID- 11065055 TI - Decrease in incidence of gestational diabetes mellitus in far north Queensland between 1992 and 1996. PMID- 11065056 TI - Breastfeeding in modern and ancient times: facts, ideas, and beliefs. PMID- 11065057 TI - Beer and breastfeeding. AB - Traditional wisdom claims that moderate beer consumption may be beneficial for initiation of breastfeeding and enhancement of breastfeeding success. Here we review the question whether or not there is any scientific basis for this popular belief. There are clear indications that beer can stimulate prolactin secretion which may enhance lactogenesis both in non-lactating humans and in experimental animals. The component in beer responsible for the effect on prolactin secretion is not the alcohol content but apparently a polysaccharide from barley, which explains that the effect on prolactin can also be induced by non-alcoholic beer. No systematic studies are available to evaluate the clinical effects of beer on induction of lactogenesis, and short term studies have shown a reduced breast milk intake by infants after moderate alcohol consumption of their mothers. It is conceivable that relaxing effects of both alcohol and components of hop might also have beneficial effects on lactogenesis is some women, but there is no hard evidence for causal effects. It appears prudent not to generally advocate the regular use of alcoholic drinks during lactation but to rather refer mothers to non-alcoholic beer, even though no adverse effects of an occasional alcoholic drink during lactation have been documented. PMID- 11065058 TI - Does breast-feeding protect against childhood obesity? AB - The impact of breast-feeding on overweight and obesity in children at school entry was assessed in a cross sectional study in Bavaria in 1997. The school entry health examination enrolled 134,577 children. Data on early feeding were collected in two rural districts (eligible population n=13,345). The analyses were confined to 5 or 6 year old children with German nationality. The main outcome measures were overweight (BMI>90th percentile for all German children seen at the 1997 school entry health examination in Bavaria) and obesity (BMI>97th percentile). Information on breast-feeding was available for 9206 children of whom 56% had been breast-fed for any length of time. In non breast fed children the upper tail of the BMI distribution was enlarged as compared to the breast-fed children whereas the median was almost identical. The prevalence of obesity in children who had never been breast-fed was 4.5% as compared to 2.8% in ever breast-fed children. A clear dose response effect for the duration of breast-feeding on the prevalence of obesity was found: 3.8%, 2.3%, 1.7% and 0.8% for exclusive breast-feeding for up to 2, 3 to 5, 6 to 12 and more than 12 months, respectively. The results for overweight were very similar. The protective effect of beast feeding on overweight and obesity could not be explained by differences in social class or lifestyle. The adjusted odds ratios of breast-feeding for any length of time was 0.71 (95% CI 0.56-0.90) for obesity and 0.77 (95%CI 0.66-0.88) for overweight. This data set did not allow to adjust for maternal weight, an important risk factor for obesity in children. Maternal overweight, however, could not explain the effect of breastfeeding on overweight and obesity in a similar study. The reduction in the risk for overweight and obesity is therefore more likely to be related to the properties of human milk than to factors associated with breast-feeding. The potential relevance of different components of human milk for the observed reduction in the risk for overweight and obesity is discussed. The preventive effect of breast-feeding on overweight and obesity is an important additional argument for the promotion of breast-feeding in industrialised countries. PMID- 11065059 TI - Nutrients, growth, and the development of programmed metabolic function. AB - For each individual, the genetic endowment at conception sets the limits on the capacity or metabolic function. The extent to which this capacity is achieved or constrained is determined by the environmental experience. The consequences of these experiences tend to be cumulative throughout life and express themselves phenotypically as achieved growth and body composition, hormonal status and the metabolic capacity for one or other function. At any time later in life the response to an environmental challenge, such as stress, infection or excess body weight is determined by an interaction amongst these factors. When the metabolic capacity to cope is exceeded, the limitation in function is exposed and expresses itself as overt disease. During early life and development the embryo, fetus and infant are relatively plastic in terms of metabolic function. The effect of any adverse environmental exposure is likely to be more marked than at later ages and the influence is more likely to exert a fundamental effect on the development of metabolic capacity. This has been characterised as "programming" and has come to be known as "the Barker hypothesis" or "the fetal origins hypothesis". Barker has shown that the size and shape of the infant at birth has considerable statistical power to predict the risk of chronic disease in later life. These relationships are graded and operate across a range of birth weight, which would generally be considered to be normal, and are not simply a feature of the extreme of growth retardation. The first evidence showed strong relations between birth weight and heart disease, the risk factors for heart disease, diabetes and hypertension, and the intermediary markers for heart disease, blood cholesterol and fibrinogen. Strong associations have also been found for bone disease, allergic disease and some aspects of brain function. In experimental studies in animals it is possible to reproduce all of the metabolic features predicted from this hypothesis by moderating the consumption of food, or its pattern during pregnancy, and determining metabolic behaviour in the offspring. It has been shown that aspects of maternal diet exert an influence on fetal growth, especially the dietary intake of carbohydrate, protein and some micronutrients. However, these relationships are less strong than might have been predicted, especially when compared with the associations which can be drawn with maternal shape, size and metabolic capacity. Maternal height, weight and body composition relate to the metabolic capacity of the mother and her ability to provide an environment in which the delivery of nutrients to the fetus is optimal. Current evidence suggests that the size of the mothers determines her ability to support protein synthesis, and that maternal protein synthesis, especially visceral protein synthesis, is very closely related to fetal growth and development. It is not clear the extent to which the effect of an adverse environment in utero can be reversed by improved conditions postnatally, but some care is needed in exploring this area, as the evidence suggests that "catch-up" growth imposes its own metabolic stress and may in itself exert a harmful effect. PMID- 11065060 TI - Early programming of glucose metabolism, insulin action and longevity. PMID- 11065061 TI - The mammary gland-infant intestine immunologic dyad. AB - The human infant has a very small immune system and needs the support of the mother with the transplacentally arrived IgG antibodies to protect tissues with inflammatogenic and energy-consuming defense. The mucous membranes, where most infections occur, need support via the specialized secretory IgA antibodies and the many other mucosal defense mechanisms provided via the mother's milk. This defense is not inflammatogenic and energy-consuming. We learn about additional defense factors in the milk, like the anti-secretory factor, which seems to protect against diarrhoea. The milk contains numerous growth factors and cytokines, like leptin, which may promote the development of the intestine as well as the immune system. Results are appearing giving interesting evidence for enhanced protection against infection also after the termination of breastfeeding. This may occur via the priming of the infant's immune system after uptake of anti-idiotypic antibodies and lymphocytes from the milk. A breastfeeding motivation study in a large Pakistani village resulted in a 50% decrease of diarrhoea and infant mortality. Deep interviews with the mothers and the traditional birth attendants suggested that even better results may be obtained. PMID- 11065062 TI - Breast feeding and the intestinal microflora of the infant--implications for protection against infectious diseases. AB - Human breast milk contains an array of factors with anti-infectious potential, such as immunoglobulins (especially secretory IgA), oligosaccharides and glycoproteins with anti-adhesive capacity, and cytokines. Breast-feeding is associated with protection from the following infections or infection-related conditions: gastroenteritis, upper and lower respiratory tract infection, acute otitis media, urinary tract infection, neonatal septicaemia and necrotizing enterocolitis. Some of the protective effects may derive from an altered mucosal colonization pattern in the breast-fed infant. In other instances breast-fed infants develop less symptoms to the same microbe which causes disease in the bottle-fed infant. An example of an altered colonization pattern is that breast fed infants have less P-fimbriated, but more type 1-fimbriated E. coli. This may protect against urinary tract infection in the breast-fed infant since P. fimbriae are the major virulence factor for urinary tract infection. An example of changed consequences of the same microbial colonization is that secretory IgA in the breast-milk protects very efficiently from translocation of intestinal bacteria across the gut mucosa by coating intestinal bacteria and blocking their interaction with the epithelium. This mechanism may protect the infant from septicaemia of gut origin and, possibly, necrotizing enterocolitis. Breast-milk is also highly anti-inflammatogenic and contains hormone like factors which counteract diarrhea. Thus, breast-fed infants may be colonized by recognized diarrheal pathogens and still remain healthy. Due to a less virulent intestinal microflora and decreased translocation breast-fed infants will obtain less stimuli for the gut immune system, resulting, in e.g., lower salivary IgA antibody titres. PMID- 11065063 TI - Opsonophagocytosis versus lectinophagocytosis in human milk macrophages. AB - Some important immunoprotective effects of human breast milk have been attributed to the presence of macrophages. We investigated the generation of superoxide anion (O2-) by monocytes and human milk macrophages after stimulation with opsonized and unopsonized zymosan in the absence and presence of mannose as an inhibitor to investigate lectinophagocytic and opsonophagocytic properties. Peripheral blood monocytes generated more O2- than human milk macrophages (417,4 + 79,1 nmol O2-/mg protein vs. 216,1 +/-15,1 nmol O2-/mg protein, p<0,05) after stimulation with opsonized zymosan. When unopsonized zymosan was used as a serum independent stimulus monocytes generated slightly less O2- in comparison to human milk macrophages (150,8 +/- 34,5 nmol/mg protein vs. 176,1 +/- 18 nmol O2-/mg protein, p<0,05). These findings demonstrate that the proportion of opsonin independent phagocytosis in human milk macrophages is higher than in monocytes (82% vs. 36%). When mannose was used as an inhibitor a significantly higher reduction of O2- generation occurred in human milk macrophages compared to monocytes stimulated with opsonized zymosan, whereas no difference was found when unopsonized zymosan was used. These results indicate that human milk macrophages are stimulated to a greater extent by opsonin-independent mechanisms than blood borne monocytes. As the colostrum and the intestinal environment of the neonate offers only a little amount of opsonins like complement and immunoglobulin G, such a differentiation to lectinophagocytic properties could bear a great advantage for protective functions of human milk macrophages. PMID- 11065064 TI - Is allergy a preventable disease? PMID- 11065065 TI - Breast-feeding and the development of cows' milk protein allergy. AB - Early feeding with cows' milk (CM) may cause cows' milk allergy (CMA). Breast milk contains many immune factors which compensate for the undeveloped defence mechanisms of the gut of the newborn infant. We studied the effect of supplementary CM feeding at the maternity hospital on the subsequent incidence of CMA, the effects of formula and breast feeding on the subsequent immunologic types of CMA, and the importance of immune factors present in colostrum in the immune responses of infants with CMA. In a cohort of 6209 infants, 824 were exclusively breast-fed and 87% required supplementary milk while in the maternity hospital: 1789 received CM formula, 1859 pasteurized human milk, and 1737 whey hydrolysate formula. The cumulative incidence of CMA, verified by a CM elimination-challenge test, was 2.4% in the CM, 1.7% in the pasteurized human milk and 1.5% in the whey hydrolysate group. Among these infants, exposure to CM at hospital and a positive atopic heredity increased the risk of CMA. Of the exclusively breast-fed infants, 2.1% had CMA. Risk factors for the development of IgE-mediated CMA were: exposure to CM at hospital, breast-feeding during the first 8 weeks at home either exclusively or combined with infrequent exposure to small amounts of CM and long breast-feeding. The content of transforming growth factor-beta1 (TGF-beta1) in colostrum from mothers of infants with IgE-mediated CMA was lower than from mothers of infants with non-IgE-mediated CMA. In infants with CMA, TGF-beta1 in colostrum negatively correlated with the result of skin prick test and the stimulation of peripheral blood mononuclear cells to CM, but positively with infants' IgA and IgG antibodies to CM proteins. Feeding of CM formula at maternity hospital increases the risk of CMA, but exclusive breast feeding does not eliminate the risk. Prolonged breast-feeding exclusively or combined with infrequent exposure to small amounts of CM during the first 8 weeks induces the development of IgE-mediated CMA. Colostral TGF-beta1 may inhibit IgE- and cell mediated reactions and promote IgG-IgA antibody production to CM in infants prone to developing CMA. PMID- 11065066 TI - Maternal asthma status alters relation of infant feeding to asthma in childhood. AB - The relation of infant feeding to childhood asthma is controversial. This study tested the hypothesis that maternal asthma alters the relation of breastfeeding to childhood asthma. Questionnaires were completed at age 6, 9 or 11 years by parents of 1043 children enrolled at birth. Active MD asthma was defined as a physician diagnosis of asthma plus asthma symptoms reported on one of the questionnaires. Duration of exclusive breastfeeding, categorized as never, < 4 months, or > or = 4 months, was based on prospective physician reports or questionnaires completed at 18 months. The relationship between breastfeeding and asthma differed by maternal asthma status. For children with maternal asthma, the percent developing active MD asthma increased significantly with longer duration of exclusive breastfeeding. Odds of developing asthma among these children were significantly elevated (OR: 5.7,CI: 2.8-11.5), after adjusting for confounders. This association of longer exclusive breastfeeding with increased risk of reported asthma among children with asthmatic mothers may be biologically based, or may reflect reporting biases. PMID- 11065067 TI - Does breast-feeding affect the risk for coeliac disease? AB - Coeliac disease, or permanent gluten sensitive enteropathy, has emerged as a widespread health problem. It is considered an immunological disease, possibly of autoimmune type, albeit strictly dependent on the presence in the diet of wheat gluten and similar proteins from rye and barley. There are reasons to believe that the aetiology of coeliac disease is multifactorial, i.e. that other environmental exposures than the mere presence in the diet of gluten affect the disease process. Our studies have shown that prolonged breast-feeding, or perhaps even more important, ongoing breast-feeding during the period when gluten containing foods are introduced into the diet, reduce the risk for coeliac disease. The amount of gluten consumed is also of importance in as much as larger amounts of gluten-containing foods increase the risk for coeliac disease, while it still is uncertain if the age for introducing gluten into the diet of infants is important. Thus, a challenging possibility, that need to be further explored, is if the coeliac enteropathy can be postponed, or possibly even prevented for the entire life span, by favourable dietary habits early in life. PMID- 11065068 TI - Breastfeeding and growth in rural Kenyan toddlers. AB - Research has not provided unequivocal support for the recommendation to continue breastfeeding until children reach at least age 24 months. In many circumstances, breastfeeding duration is chosen or conditioned by factors other than scientific evidence and recommendations. Even in communities where breastfeeding into the second year is the norm, a significant number of toddlers are weaned before the recommended age. The research reported here was conducted in a rural community of western Kenya. We prospectively followed a cohort of 264 children for 6 months (mean age at baseline, 14.1 +/- 2.4 months) to examine the effect of variable breastfeeding duration on length and weight gain. We found that breastfeeding was positively associated with growth in a manner that we inferred to be causal, the effect being stronger on linear growth than on weight gain. This was despite the fact that in a cohort where 95% were breastfeeding at baseline, the prevalence of stunting (height-for-age below -2 standard deviations of the WHO-NCHS reference) was already 48%. The present paper examines the socioeconomic characteristics, sanitation, morbidity, and complementary feeding practices that define the context of this apparently contradictory relationship. The population was poor, no household had running water, and malaria is endemic in the study area. Complementary feeding was initiated for 93% of the cohort before age 3 months. The weaning diet was bulky (77% energy from carbohydrate), and high in phytate content ([phytate]:[zinc] molar ratio, 28). Diet quality, judged by diversity and animal source food intake, was low. Several micronutrient intakes were below current recommendations, including riboflavin (63%), niacin equivalents (64%), calcium (72%), iron (74%) and zinc (33%). Based on a locally defined socioeconomic status scale, children in higher SES households were breastfed for a shorter duration than were children from poorer households. Sanitation and water consumption modified the effect of breastfeeding duration on growth: the effect was stronger in the absence of a pit latrine and at low water consumption. Our results support the recommendation to sustain breastfeeding in the second year, particularly in economically depressed environments with inadequate sanitation and water supplies. PMID- 11065069 TI - Breastfeeding and stunting among toddlers in Peru. PMID- 11065070 TI - Breastfeeding and growth in rural Senegalese toddlers. PMID- 11065071 TI - Duration of breast-feeding and linear growth. PMID- 11065072 TI - The association between prolonged breastfeeding and poor growth--what are the implications? AB - The smaller size of breast fed children in infancy and thereafter in malnourished and well-nourished populations has resulted in rushes to judgement that have been shown to be ill-advised. The reasons for the smaller size in malnourished populations is due to retaining the small and sickly child at the breast (reverse causality) and the consequent continuing sickliness of this breast fed child (negative confounding). Once the reverse causality and negative confounding have been taken into account breast feeding improves growth, at least through the second year of life. Thus prolonged breastfeeding should always be fostered, especially in malnourished populations. An exception remains when breast milk may transmit disease to the suckling child. In well-nourished populations the magnitude of the difference between breast fed and weaned children is much less than in malnourished populations, is observed to increase over the first year of life, but to have disappeared by the end of the second year. One may never-the less be concerned that complimentary feeding practices are not adequate for these children. PMID- 11065073 TI - Breastfeeding and HIV-1 infection. A review of current literature. PMID- 11065074 TI - Subclinical mastitis as a risk factor for mother-infant HIV transmission. AB - Subclinical mastitis, as diagnosed by an elevated sodium/potassium ratio in milk accompanied by an increased milk concentration of the inflammatory cytokine, interleukin-8 (IL8), was found to be common among breast feeding women in Bangladesh and Tanzania. Subclinical mastitis results in leakage of plasma constituents into milk, active recruitment of leukocytes into milk, and possible infant gut damage from inflammatory cytokines. Therefore, we wished to investigate whether subclinical mastitis was related to known risk factors for postnatal mother-to-child HIV transmission, that is, high milk viral load or increased infant gut permeability. HIV-infected South African women were recruited at the antenatal clinic of McCord's Hospital, Durban. Risks and benefits of different feeding strategies were explained to them and, if they chose to breast feed, they were encouraged to do so exclusively. Women and infants returned to the clinic at 1, 6 and 14 weeks postpartum for an interview about infant health and current feeding pattern, a lactulose/mannitol test of infant gut permeability, and milk sample collection from each breast separately for analysis of Na/K ratio, IL8 concentration and viral load in the cell-free aqueous phase. Only preliminary cross-sectional analyses from an incomplete database are available at this point. Moderately (0.6-1.0) or greatly (>1.0) raised Na/K ratio was common and was often unilateral, although as a group right and left breasts did not differ. Considering both breasts together, normal, moderately raised or greatly raised Na/K was found, respectively, in 51%, 28%, 21% of milk samples at 1 week (n=190); 69%, 20%, 11% at 6 weeks (n=167); and 72%, 16%, 12% at 14 weeks (n=122). IL8 concentration significantly correlated with both Na/K and viral load at all times. Na/K correlated with viral load at 1 and 14, but not 6 weeks. At 1 and 14 weeks, geometric mean viral loads in samples with Na/K > 1.0 were approximately 4 times those in samples with Na/K < 0.6. At 1 week but not later times, exclusive breast feeding was associated with lower milk viral load than was mixed feeding. Gut permeability was unrelated to milk Na/K ratio or IL8 concentration and was not significantly increased by inclusion of other foods than breast milk in the infant's diet. The results suggest that subclinical mastitis among HIV-infected women may increase the risk of vertical transmission through breast feeding by increasing milk viral load. The importance of various causes of subclinical mastitis, which likely differ at 1 week from at later times and may include local infection or sterile inflammation, systemic infection, micronutrient deficiencies, or poor lactation practices, needs to be further clarified so that appropriate interventions can be implemented. PMID- 11065075 TI - Recommendations on feeding infants of HIV positive mothers. WHO, UNICEF, UNAIDS guidelines. PMID- 11065076 TI - Transmission of cytomegalovirus infection through breast milk in term and preterm infants. The role of cell free milk whey and milk cells. AB - We investigated the reactivation of cytomegalovirus during lactation and analysed the role of human milk whey and milk cells in mother-to-child-transmission. In contrast to term infants, preterm infants may be infected symptomatically by breastfeeding. Human milk whey is the material of choice for detection of maternal DNAlactia and virolactia, whereas milk cells not necessarily have to be infected in transmitters. PMID- 11065077 TI - Physiology of oligosaccharides in lactating women and breast fed infants. PMID- 11065078 TI - Immunomodulatory effects of breast milk oligosaccharides. AB - Breast milk oligosaccharides are excreted in urine in amounts that suggest that they may exist in the circulation at levels compatible with a physiological function. Some oligosaccharides have structural similarity to cellular adhesion molecules and may influence adhesion of cells in breast fed infants. In this study, breast milk oligosaccharides were purified and incubated in assays of cell adhesion. They were found to inhibit neutrophil adhesion to stimulated vascular endothelial cells in a dose dependent fashion. In contrast they enhanced platelet neutrophil complex formation. These results indicate that breast milk oligosaccharides may play a physiological role in modulating cellular adhesion in vivo. PMID- 11065079 TI - Polyunsaturated fatty acid supply with human milk. Physiological aspects and in vivo studies of metabolism. AB - The origin of polyunsaturated fatty acids (PUFA) in human milk has not been studied in detail. Diet, liberation from maternal stores and endogenous synthesis from precursors may contribute to PUFA present in human milk. Other factors influencing lipid content and fatty acid composition such as gestational age, stage of lactation, nutritional status and genetical background are known. In a series of in vivo studies using stable isotope methodologies we investigated the metabolism of PUFA during lactation. With this techniques the transfer of single dietary fatty acids into human milk, the oxidation and the deposition in tissues were estimated. Our studies demonstrate that the major part of PUFA in human milk seems not to be derived directly from the maternal diet but from body stores. Nevertheless diet is important, because long term intakes affect composition of body stores. PMID- 11065080 TI - Environmental exposure to polychlorinated biphenyls (PCBs) and dioxins. Consequences for longterm neurological and cognitive development of the child lactation. AB - Polychlorinated biphenyls (PCBs) and dioxins are environmental pollutants. Prenatally, as well as postnatally through breast feeding, large amounts are transferred from mother to the child. Formula is free of these substances. Considering their potential developmental neurotoxicity, we investigated long term effects of perinatal exposure to PCBs and dioxins on neurological and cognitive development. Given the evidence that PCBs exert oestrogenic effects, and oestrogens are known to suppress lactation, we investigated the effect of maternal PCB body load on lactation performances as well. METHODS: A group of 418 infants were followed from birth up to 6 years of age. Half of them were fully breast fed (BF) for at least 6 weeks. Prenatal PCB exposure was measured from cord and maternal blood. Postnatal exposure was reflected by PCB and dioxin levels in breast and formula milk and plasma PCB levels at 42 months of age. Both neurological and cognitive development were taken as outcome variable at 18, 42 months and at 6 years of age. At 18 and 42 months of age neurological condition was evaluated according to Hempel and at 6 years of age according to Touwen. Condition was evaluated in terms of optimality. Separately, the fluency of movements was scored. Cognitive abilities were measured at 18 months by the Bayley Scales of Infant Development, at 42 months of age by the Kaufman Assessment Battery for Children (K-ABC) and at 6 years of age by the McCarthy Scales. Daily breast milk volume and milk fat content in relation to PCB body load was evaluated in 102 mothers. Multivariate regression models were applied to analyse associations of measured exposure variables with independent variables adjusted for confounders. RESULTS: At 18 months of age cognitive development was not affected by either pre- or postnatal exposure to the measured PCBs and dioxins. However, neurological examination showed an adverse effect of prenatal exposure to the measured pollutants on neurological optimality score. At 42 months of age we found negative associations between prenatal PCB exposure on cognitive development. However no effect was demonstrated on postnatal exposure to the measured pollutants. Neurological development was not affected by either pre- or postnatal exposure to PCBs and dioxins. At 6 years of age the preliminary results revealed evidence that cognitive development is affected by prenatal exposure to these pollutants in children from young mothers. An adverse effect of prenatal exposure on neurological outcome was also demonstrated in the formula fed group but not in the breast fed group. Despite a higher PCB exposures from breast milk we found at 18 months, 42 months of age, and at 6 years of age a beneficial effect of breast feeding on the quality of movements, in terms of fluency, and on the cognitive development tests. Maternal PCB body load was inversely related to 24-h breast milk volume and milk fat content. CONCLUSION: These data give evidence that prenatal exposure to PCBs do have subtle negative effects on neurological and cognitive development of the child up to school-age. Human breast milk volume and fat content is adversely affected by the presently encountered PCB levels in W. Europe. Our studies showed evidence that breast feeding counteracts the adverse developmental effects of PCBs and dioxins. PMID- 11065081 TI - Transition of nitro musks and polycyclic musks into human milk. AB - Synthetic musks are widely used in various consumer products. The identification of nitro musks in human milk in the early 1990s in connection with evidence for cancerogenicity in animal experiments have caused public concern. However, the validity of previously reported quantitative data has been questioned. Polycylic musks have hardly been investigated so far. The present study aimed at providing accurate current data on the occurrence of nitro and polycyclic musks in human milk. Samples from 40 healthy breast feeding mothers were analysed under carefully controlled conditions avoiding secondary contamination. As in earlier studies, among the nitro compounds musk xylene and ketone were the most frequently detected substances. However, much lower concentrations (roughly by a factor of 10) were found (musk xylene: median 6.1 ng/kg fat). Among the polycylic musks HHCB was found in most samples (median 64 ng/kg fat). Scientific knowledge on possible routes of exposure and health risk aspects is summarized and discussed. PMID- 11065082 TI - Exposition to and health effects of residues in human milk. AB - A great variety of drugs, cosmetics, food ingredients as well as environmental contaminants are secreted with human milk as a result of actual exposure or the accumulated body burden of the mother. Of great concern and least amenable to short-term intervention are persistent substances in the environment with long half-lives in the body due to their lipophilic properties and minimal degradation. Polyhalogenated aromatic hydrocarbons, namely organochlorine pesticides, polychlorinated biphenyls (PCB) and polychlorinated dibenzodioxins (PCDD) and dibenzofurans (PCDF) are fetotoxic, neurotoxic, immunotoxic, some are promoting carcinogens and/or interfere with hormonal receptors. They pass the placenta and equilibrate among the lipid compartments of the body including breast milk lipids. Transplacental exposure is more relevant with regard to physical development and cognitive functioning of the child than postnatal exposure via breastmilk. Restrictions for production, use and release have been successful in decreasing exposure as shown by a downward trend of their contents both in human milk and serum lipids for the last 15 to 20 years. It is difficult to evaluate the potentially late effects of the exposure via breastmilk which is 10 to 100 times higher in industrialised countries than the tolerable daily intake (TDI) of 1 to 4 toxic equivalents (WHO-TEQ) pg/kg/day established in 1998 by WHO for dioxins and dioxin-like PCBs but which lasts for 0.6% of the expected life span only. Carefully conducted long-term follow-up of cohorts with defined exposure levels, with consideration of numerous biological and psychological parameters, is expected to provide the answer. PMID- 11065083 TI - Promotion of breastfeeding intervention trial (PROBIT): a cluster-randomized trial in the Republic of Belarus. Design, follow-up, and data validation. AB - This paper summarizes the objectives, design, follow-up, and data validation of a cluster-randomized trial of a breastfeeding promotion intervention modeled on the WHO/UNICEF Baby-Friendly Hospital Initiative (BFHI). Thirty-four hospitals and their affiliated polyclinics in the Republic of Belarus were randomized to receive BFHI training of medical, midwifery, and nursing staffs (experimental group) or to continue their routine practices (control group). All breastfeeding mother-infant dyads were considered eligible for inclusion in the study if the infant was singleton, born at > or = 37 weeks gestation, weighed > or = 2500 grams at birth, and had a 5-minute Apgar score > or = 5, and neither mother nor infant had a medical condition for which breastfeeding was contraindicated. One experimental and one control site refused to accept their randomized allocation and dropped out of the trial. A total of 17,795 mothers were recruited at the 32 remaining sites, and their infants were followed up at 1, 2, 3, 6, 9, and 12 months of age. To our knowledge, this is the largest randomized trial ever undertaken in area of human milk and lactation. Monitoring visits of all experimental and control maternity hospitals and polyclinics were undertaken prior to recruitment and twice more during recruitment and follow-up to ensure compliance with the randomized allocation. Major study outcomes include the occurrence of > or = 1 episode of gastrointestinal infection, > or = 2 respiratory infections, and the duration of breastfeeding, and are analyzed according to randomized allocation ("intention to treat"). One of the 32 remaining study sites was dropped from the trial because of apparently falsified follow-up data, as suggested by an unrealistically low incidence of infection and unrealistically long duration of breastfeeding, and as confirmed by subsequent data audit of polyclinic charts and interviews with mothers of 64 randomly selected study infants at the site. Smaller random audits at each of the remaining sites showed extremely high concordance between the PROBIT data forms and both the polyclinic charts and maternal interviews, with no evident difference in under- or over-reporting in experimental vs control sites. Of the 17,046 infants recruited from the 31 participating study sites, 16,491 (96.7%) completed the study and only 555 (3.3%) were lost to follow-up. PROBIT's results should help inform decision-making for clinicians, hospitals, industry, and governments concerning the support, protection, and promotion of breastfeeding. PMID- 11065084 TI - Provision of supplementary fluids to breast fed infants and later breast feeding success. AB - It has been shown that altering hospital policies in a way to avoid interference of routine prescriptions with initiation of breast feeding and to provide active encouragement to mothers and personnel can result in significant benefit for later breast feeding success. It is less clear, however, which of the elements of a promotional programme such as UNICEF/WHO's "ten steps to successful breast feeding" are absolutely essential and which can be adapted to local cultural habits. We performed an open randomized multicenter study in Switzerland to evaluate, whether restriction of supplementary fluids for breast fed infants in the first week of life and strict avoidance of artificial teats and pacifiers affects later breast feeding success. Follow up to 6 months was ensured by mailed questionnaires. 602 mother infant pairs were enrolled. Of 294 infants in the intervention group 39% were excluded from the final analysis because of protocol violations, mainly maternal request for the use of pacifiers or bottles. Though the number of dextrin maltose supplements during the first two days (1.7 vs. 2.2 on day 1, 2.2 vs. 2.6 on day 2) and the percentage of infants receiving any supplement (85% vs. 96.6%) was significantly smaller in the intervention group, the difference was disappointingly small. The prevalence of breast feeding was 100% vs. 99% at day 5, 88% vs. 88% at 2 months, 75% vs. 71% at 4 months and 57% vs. 55% at 6 months, none of the differences being significant. We conclude that rigorous adherence to all of the ten steps may encounter obstinate resistance from cultural habits even in a population highly favourable to breast feeding. An improvement in adherence does not necessarily lead to better breast feeding success. The results of the few comparable studies in the literature show also that cultural practices during the first months of life may influence profoundly the long term effects of interventions during the first days of life. PMID- 11065085 TI - Breastfeeding promotion--is its effectiveness supported by scientific evidence and global changes in breastfeeding behaviors? PMID- 11065086 TI - Apoptosis in lactating rat mammary tissue using TUNEL method. PMID- 11065087 TI - Energy intake and growth of breast-fed infants in two regions of Mexico. PMID- 11065088 TI - Effect of human milk and recombinant EGF, TGFalpha, and IGF-1 on small intestinal cell proliferation. PMID- 11065089 TI - Low breast milk vitamin A concentration reflects an increased risk of low liver vitamin A stores in women. PMID- 11065090 TI - Vitamin A in milk can potentially reduce the replication of enveloped viruses in infants. PMID- 11065091 TI - Nucleoside analyses of human milk at 4 stages of lactation. PMID- 11065092 TI - Quantitative analysis of human milk oligosaccharides by capillary electrophoresis. PMID- 11065093 TI - Zinc intakes and plasma concentrations in infancy. PMID- 11065094 TI - Nutritive significance of element speciation in breast milk. The case of calcium, copper, iron, magnesium, manganese, and zinc. PMID- 11065095 TI - Human milk mercury (Hg) and lead (Pb) levels in Vienna. PMID- 11065096 TI - Breastfeeding and atopic sensitisation. PMID- 11065097 TI - Cytokine production by leukocytes from human milk. PMID- 11065098 TI - Breastfeeding and asthma in children. A prospective cohort study. PMID- 11065099 TI - Docosahexaenoic acid (DHA) status of breastfed malnourished infants and their mothers in North Pakistan. PMID- 11065100 TI - Long term effect of breast feeding on essential fatty acid status in healthy, full-term infants. PMID- 11065101 TI - Human milk fatty acid profiles from Australia, Canada, Japan, and The Philippines. PMID- 11065102 TI - Short-and long term variation in the production, content, and composition of human milk fat. PMID- 11065103 TI - Dietary fish and the docosahexaenoic acid (DHA) content of human milk. PMID- 11065104 TI - Fatty acid composition of mature breast milk according to the mothers diet during pregnancy. AB - Mothers taking 200 g of fish per week, showed a greater content in mature breast milk of n-3 fatty acids, particularly DHA. AA was not decreased. Vitamin D content was low despite mothers were living in a sunny and temperate area. The content of 25 hydroxyvitamin D is increased in the group on fish intake, probably pointing out its marine source. PMID- 11065105 TI - Contribution of dietary and newly formed arachidonic acid to milk secretion in women on low fat diets. PMID- 11065106 TI - 13C-linoleic acid oxidation and transfer into milk in lactating women with contrasting body mass index. PMID- 11065107 TI - Arachidonic (AA) and docosahexaenoic (DHA) acid content in healthy infants fed with an HA milk formula supplemented with LCPUFA and in breast fed infants. PMID- 11065108 TI - Low contribution of docosahexaenoic acid to the fatty acid composition of mature human milk in Hungary. PMID- 11065109 TI - Malnourished mothers maintain their weight through out pregnancy and lactation. Results from Guatemala. PMID- 11065110 TI - Effect of exercise and energy restriction on leptin during lactation. PMID- 11065111 TI - Food intakes in a group of breast-feeding and not breast-feeding mothers. AB - The relation between mother's diet and breast milk composition is still an open question. An important issue is whether mothers who are breast feeding modify, respect to before lactation, food intake to satisfy the increased requirements. PMID- 11065112 TI - Illness-induced anorexia in the breast-fed infants. Role of IL-1beta and TNF alpha. PMID- 11065113 TI - Maternal perception of the onset of lactation: a valid indicator of lactogenesis stage II? PMID- 11065114 TI - The onset of lactation: implications for breast-feeding promotion programs. PMID- 11065115 TI - Pre-term delivery and breast expression: consequences for initiating lactation. PMID- 11065116 TI - Breastfeeding rates of VLBW infants--influence of professional breastfeeding support. PMID- 11065117 TI - Breastfeeding in Gent, Belgium. A cohort study on breastfeeding rate and duration. PMID- 11065118 TI - Breast-feeding pattern and influencing factors in Lithuania. PMID- 11065119 TI - Factors influencing a mother's decision to breastfeed. PMID- 11065120 TI - Use of soft laser in the therapy of sore nipples in breastfeeding women. PMID- 11065121 TI - Natural feeding of premature infants. PMID- 11065122 TI - News about human milk banking in Germany. PMID- 11065123 TI - Neuroprotection by group I metabotropic glutamate receptor antagonists in forebrain ischemia of gerbil. AB - Stimulation of group I metabotropic glutamate receptors (mGluR 1 and 5) activates G-protein coupled-phospholipase C (PLC) to release 1,2-diacylglycerol (DAG) and arachidonic acid (ArAc). To elucidate the role of group I mGluR, we tested the effects of (S)-alpha-methyl-4-carboxy-phenylglycine (MCPG, mGluR 1 and 5 antagonist), 1-aminoindan-1,5-dicarboxylic acid (AIDA, mGluR 1a specific antagonist) and 2-methyl-6-(phenylethynyl) pyridine (MPEP, mGluR 5 antagonist) on ArAc release and neuronal survival after transient forebrain ischemia in gerbils. Ischemia resulted in (a) significant release of ArAc at 1-day reperfusion and (b) significant neuronal death in the hippocampal CA1 subfield after 6-day reperfusion. MCPG and MPEP decreased ArAc release and also significantly increased neuronal survival. AIDA was less effective in decreasing ArAc release and had no effect on neuronal death. These results suggest that activation of mGluR 5 may be an important pathway in ArAc release and neuronal death after transient ischemia. PMID- 11065124 TI - Possible involvement of light regulated gonadotropin-releasing hormone neurons in biological clock for reproduction in the cerebral ganglion of the ascidian, Halocynthia roretzi. AB - Since ascidians, a primitive chordate, spawn at a fixed latency after sunrise, light must regulate a biological clock for reproduction in the ascidians. A retinal protein found in the cerebral ganglion of the ascidian is a candidate for the photoreceptor that might drives the change in gonadal activity via the gonadotropin-releasing hormone (GnRH) system. Photoresponses of the cerebral ganglion of ascidian, Halocynthia roretzi, were examined and two light-evoked responses recorded extracellularly, a light-evoked slow potential and light inhibition of high frequency spontaneous discharges. These results suggest that pacemaker signals of GnRH neurons might be regulated by photoreceptor activation. Immunohistochemical studies showed photoreceptor cells located close to the GnRH neurons and thus the photosignal might proceed from photoreceptor cell to GnRH neuron intercellularly. PMID- 11065125 TI - Effects of systemic injection of interleukin-1beta on gastric vagal afferent activity in rats lacking type A cholecystokinin receptors. AB - We have shown that systemic administration of interleukin-1beta (IL-1beta) excites gastric vagal afferent activity in part via stimulation of type A cholecystokinin (CCK-A) receptors in rats. The present study was undertaken to determine whether the response of the gastric vagal afferent nerve to systemic IL 1beta is altered in Otsuka Long-Evans Tokushima Fatty (OLETF) rats, which lack CCK-A receptors. The response was compared with that of the control strain, Long Evans Tokushima Otsuka (LETO) rats. All animals were anesthetized with pentobarbital and artificially ventilated. Intravenous administration of 4 microg/kg of IL-1beta increased gastric vagal afferent activity in both LETO and OLETF rats, whereas a smaller dose of 2 microg/kg of IL-1beta increased activity only in the OLETF rats. The present results demonstrate that the response of the gastric vagal afferent activity in CCK-A receptor deficient OLETF rats was more sensitive to intravenous administration of IL-1beta than was in control LETO rats. PMID- 11065126 TI - Expression of integrin alpha2beta1 in axons and receptive endings of neurons in rat, hairy skin. AB - Integrin alpha2beta1 has been considered as a mechano-chemical transducer in endothelial and muscle cells. However, little data is available to show whether integrins play a role in the process of mechanical transduction in peripheral mechanosensory neurons. Using immunohistochemistry, we demonstrate that cutaneous neurons express the extracellular matrix (ECM) receptor integrin alpha2beta1. Specifically, we show that integrins alpha2 and beta1 are co-localized with peripherin in the receptive endings of cutaneous neurons in rat, hairy skin. Integrin immunofluorescence was minimal along the axons of large diameter neurons. These results, together with findings by other investigators, provide evidence suggesting that integrin alpha2beta1 may be a linking agent between mechanical stress in the ECM and modulation of the neuronal response of mechanically sensitive neurons. PMID- 11065127 TI - Event-related potential N270 is elicited by mental conflict processing in human brain. AB - We recorded event-related potentials (ERPs) in 15 subjects in order to elicit a N270 of arithmetic conflict. Subjects calculated an arithmetic problem and matched their calculation result to an answer digit. They pressed a button when the presented digit is a true answer (condition 1) and pressed another button when the answer is false (condition 2). ERP components of P90, N130, P180, N200 and late positive component (LPC) were recorded in condition 1. In condition 2, N270 was elicited between N200 and LPC and it peaked at approximately 270 ms (268.6 +/- 29.0 ms at Cz). The peak latency of LPC in condition 2 (405.7 +/- 51.3 ms) is significantly delayed than condition 1 (307.5 +/- 22.7 ms). N270 reflects the endogenous conflict processing in human brain. PMID- 11065128 TI - Different responses of ventromedial hypothalamic neurons to leptin in normal and early postnatally overfed rats. AB - Leptin is crucially involved in the central nervous regulation of body weight. Neurons of the ventromedial hypothalamic nucleus (VMH) express leptin receptors and signal satiety with increase in their firing. Normally, leptin mainly activates VMH neurons. Rats grown up in small litters (SL) develop persistent hyperphagia and obesity throughout life. We studied single unit activity in hypothalamic brain slices of juvenile SL rats overweight due to early postnatal overfeeding (Mann-Whitney U-test, P < 0.001). VMH neurons of normal rats were mainly activated by leptin (Wilcoxon test, P < 0.05, n = 39), whereas neurons of overweight SL rats were mainly inhibited (Wt, P < 0.001, n = 33). This clearly altered response to leptin in neonatally overnourished rats might contribute to their persistent overweight throughout life. PMID- 11065129 TI - Immunohistochemical study on the distribution of the type I and type II voltage gated sodium channels in the gerbil cerebellum. AB - In this study, we investigated the distribution of the type I and type II Na+ channels in the gerbil cerebellum by immunohistochemistry. Strong uniform staining for type I was observed in the granular layer, whereas there was little evidence of concentrated labeling in the cell bodies and processes of Purkinje cells. The most intense staining for type II was observed in the cell bodies and dendrites of Purkinje cells, with a strong signal in the molecular layer. This localization study has shown clearly that the type I and type II Na+ channel subunits have differential distribution in the gerbil cerebellum, for the first time. The present study may provide useful data for the future investigations to understand the roles of voltage-gated sodium channels in neurological pathways. PMID- 11065130 TI - A FE65 polymorphism associated with risk of developing sporadic late-onset alzheimer's disease but not with Abeta loading in brains. AB - The FE65 protein was previously described interacting with amyloid protein precursor (APP) and mediating its internalization. Hu et al. (Hum. Genet., 103 (1998) 295) recently reported that a deletion polymorphism in intron 13 of the FE65 gene may be protective for sporadic Alzheimer's disease (AD) forms and suggested that this deletion may modify splicing between exon 13 and 14 (the two exons encoding the interaction domain of FE65 with APP). We tested the impact of this polymorphism in 646 controls and 639 sporadic AD cases. We were only able to detect a protective effect of the deletion in the population over 75 years (odds ratio = 0.53, 95% confidence interval (0.35-0.82), P= 0.002). Furthermore, no association of this polymorphism with Abeta40, Abeta42(43) and total Abeta loads were detected in 74 AD brains, although, we could expect that this deletion was associated with modifications of the APP metabolism. In conclusion, the FE65 gene may be a minor genetic determinant only for sporadic late-onset AD forms, although, we cannot conclude that this impact is mediated by a modulation of the APP process and/or Abeta peptide deposition. PMID- 11065131 TI - Cyclin-dependent kinase 5 colocalizes with phosphorylated tau in human inclusion body myositis paired-helical filaments and may play a role in tau phosphorylation. AB - To investigate the possible role of cyclin-dependent kinase 5 (cdk5) in the formation of paired helical filaments (PHFs) in muscle of patients with inclusion body myositis (IBM), we immunolocalized cdk5, by light- and electron- microscopy, in muscle biopsies of six IBM patients. Approximately 80-90% of IBM vacuolated muscle fibers, and 10-15% of nonvacuolated fibers, contained well defined cdk5 immunoreactive inclusions that colocalized with phosphorylated tau in 70-80% of those fibers. Immunoelectronmicroscopy revealed the association of cdk5 with tau immunoreactive PHFs. In all biopsies that contained them, regenerating muscle fibers had diffuse, moderate to strong cdk5 immunoreactivity. At all neuromuscular junctions, there was strong cdk5 immunoreactivity postsynaptically. Our study suggests that cdk5: (1) plays a role in IBM pathogenesis, possibly mediating phosphorylation of PHF-related tau; (2) is involved in muscle regeneration; and (3) has a novel function at normal neuromuscular junctions. PMID- 11065132 TI - Nerve growth factor-mediated expression of galectin-3 in mouse dorsal root ganglion neurons. AB - Galectin-3, a member of the galectin family of beta-galactoside-specific lectins has been found to be expressed by subsets of dorsal root ganglion (DRG) neurons during development and in adulthood. Here we show that (i) after 3-7 days in vitro, DRG neurons derived from neonatal mice express galectin-3 intra- and extracellularly and (ii) lectin expression requires the presence of nerve growth factor (NGF). After 3 days in vitro, a higher number of DRG neurons expressed galectin-3 in the presence of NGF (65 +/- 7%) than in the presence of brain derived neurotrophic factor (BDNF, 30 +/- 3%) or neurotrophin-3 (NT-3, 34 +/- 3%). After 7 days in vitro, these numbers dropped to 51 +/- 3% (for NGF), 0% (for BDNF) and 8 +/- 4% (for NT-3), respectively. Our findings provide first evidence for the contribution of a neurotrophin to the neuronal expression of galectins and suggest an NGF/TrkA-mediated expression of galectin-3 by early postnatal DRG neurons. PMID- 11065133 TI - Patterns of dopamine overflow in mouse nucleus accumbens during intracranial self stimulation. AB - Dopamine (DA) overflow in the mouse nucleus accumbens during intracranial self stimulation (ICSS) of the median forebrain bundle was estimated by chronoamperometry with removable carbon fibre electrodes. The specificity of the voltammetric signal was confirmed pharmacologically. The parameters of stimulation (50 Hz, 0.5 s train length) allowed us to obtain measurable DA release and to maintain ICSS. Continuous (CR) and fixed-ratio (FR8) schedule of reinforcement showed differing correspondence of the patterns of DA release with the patterns of stimulation/nose-poking. The CR schedule induced a high rate nose poking and tonic increase in dopamine overflow, which became decreased following the first periods of self-stimulations. The FR schedule induced stable peaks of DA overflow during the entire period of ICSS. We conclude that the availability of a readily-releasable pool of DA in presynaptic terminals determined the pattern of dopamine overflow in the nucleus accumbens during ICSS in mice. PMID- 11065134 TI - Modification of cardiovascular response of adenosine A2 receptor agonist by adenylate cyclase in the spinal cord of rats. AB - This study was performed to investigate the influence of spinal adenosine A2 receptors on the central regulation of blood pressure (BP) and heart rate (HR), and to define whether its mechanism is mediated by adenylate cyclase or guanylate cyclase. Intrathecal (i.t.) administration of drugs at the thoracic level were performed in anesthetized, artificially ventilated male Sprague-Dawley rats. Injection (i.t.) of adenosine A2 receptor agonist, 5'-(N-cyclopropyl) carboxamidoadenosine (CPCA; 1, 2 and 3 nmol) produced a dose dependent decrease of BP and HR. Pretreatment with adenylate cyclase inhibitor, MDL-12,330, attenuated the depressor and bradycardiac effects of CPCA (2 nmol), but not with guanylate cyclase inhibitor, LY-83,583. These results suggest that adenosine A2 receptor in the spinal cord plays an inhibitory role in the central cardiovascular regulation and that the depressor and bradycardiac actions are mediated by adenylate cyclase. PMID- 11065135 TI - Co-induction of heme oxygenase-1 and peroxiredoxin I in astrocytes and microglia around hemorrhagic region in the rat brain. AB - Heme[none1] oxygenase-1 (HO-1) and peroxiredoxin I (PrxI) are known to be oxidative stress- and heme-related proteins. The antioxidant activity of PrxI is inhibited by heme, therefore co-expression of HO-1 and PrxI is considered to be a reasonable mechanism to maintain its antioxidative function. Immunoblotting demonstrated that HO-1 and PrxI were induced around the hemorrhagic region. Immunohistochemical studies revealed that, in acute phase, HO-1 and PrxI were induced primarily in microglia. In the subacute and chronic phase, the immunoreactivity of HO-1 and PrxI in astrocytes was the most intense. These data are the first to demonstrate co-induction of HO-1 and PrxI in the brain. Our results suggest that HO-1 and PrxI are localized in a similar manner to assure the antioxidant activity of PrxI under stress conditions associated with intracerebral hemorrhage. PMID- 11065136 TI - Expression of brain/kidney protein in Muller cells of rat retina following transient ischemia. AB - We have investigated the expression and cellular localization of brain/kidney (B/K) protein in the rat retina following transient ischemia. In the normal retina, strong B/K immunoreactivity was localized to some ganglion cells. In addition, a few radial Muller cell processes showed B/K immunoreactivity. Following ischemia and reperfusion, most B/K-labeled ganglion cells were lost, whereas between 1 day and 2 weeks post-lesion B/K immunoreactivity appeared in many more Muller cell processes with increasing intensity. Quantitative evaluation by immunoblotting confirmed that B/K expression then decreased progressively, to 35% of control values at four weeks post-lesion. Our findings suggest that Muller cells are involved in the pathophysiology of retinal ischemia through the expression of B/K following transient ischemia. PMID- 11065137 TI - Detection of single-stranded DNA in the hydropic vestibule after the direct injection of antigen into the endolymphatic sac of guinea pigs. AB - Immunohistochemical study for single-stranded DNA (ssDNA) with vestibule of guinea pigs was performed after the injection of keyhole limpet hemocyanin (KLH) into the right endolymphatic sac. Endolymphatic hydrops became evident by expansion of the Reissner's membrane in the cochlea of all animals 1 day after the injection of KLH. Increased ssDNA expression was detected in the sensory epithelium and transitional area, while temporal bones in the control group did not show any ssDNA immunoreactivities. ssDNA is accompanied with the apoptotic change in the vestibule. Our results suggest that apoptotic changes could be involved in the hydropic vestibule and these phenomena lead inner ear disturbance as seen in endolymphatic hydrops. PMID- 11065138 TI - Immunoreactivity for glutamic acid decarboxylase and N-methyl-D-aspartate receptors of intracellularly labeled respiratory neurons in the cat. AB - In adult cats, immunofluorescence images of glutamic acid decarboxylase (GAD) and N-methyl-D-aspartate (NMDA) receptors were achieved in the ventral respiratory group (VRG) neurons, which had been individually identified by in vivo intracellular recording and labeling with neurobiotin. Among augmenting inspiratory (aug-I), postinspiratory (post-I), and augmenting expiratory (aug-E) neurons labeled, GAD-immunoreactivity was demonstrated only in those neurons that were not antidromically activated (NAA) by stimulation of the vagus nerve and the C2-C3 spinal cord. Substantial immunoreactivity for NMDA receptors was presented in virtually all types of neurons, but lesser reactivity in aug-E bulbospinal neurons. These results suggest that the aug-I, post-I, and aug-E types of NAA neurons are gamma-aminobutyric acid (GABA)ergic and that NMDA receptors distribute in lesser degree in aug-E bulbospinal neurons than in other types of VRG neurons. PMID- 11065139 TI - Expression of metallothionein-III mRNA and its regulation by levodopa in the basal ganglia of hemi-parkinsonian rats. AB - In the brain, metallothionein (MT)-III exhibits a free radical scavenging activity. Here we examined the expression of MT-III mRNA in the basal ganglia of 6-hydroxydopamine (6-OHDA)-lesioned hemi-parkinsonian rats and its regulation by levodopa. The level of MT-III mRNA was significantly decreased in the striatum of 6-OHDA-lesioned side. Levodopa treatment significantly increased the expression of striatal MT-III mRNA in the non-lesioned side, but showed no significant effect in the 6-OHDA-lesioned side. These results suggest that the regulation of MT-III mRNA may be related to the progressive degeneration in parkinsonism. PMID- 11065140 TI - Visual information processing in patients with schizophrenia: evidence for the impairment of central mechanisms. AB - Patients with schizophrenia are especially impaired in the detection of spatial location if the briefly presented target stimulus is followed by a mask in a close temporal proximity (target location backward masking (BM) paradigm). It has been suggested that this phenomenon is related to the impairment of low spatial and high temporal frequency-sensitive transient (magnocellular) visual channels. To test this hypothesis, we measured target location BM and visual contrast sensitivity (CS) in clinically remitted patients with schizophrenia. In the BM task, subjects were asked to indicate the position of letters appearing at four possible spatial locations. In the CS test, a two-alternative forced choice method was used to measure the minimal contrast level required for the detection of horizontal gratings set at low spatial and high temporal frequencies (0.5 cycle/degree and 8 Hz, respectively). We found that the schizophrenia patients with normal CSs (spared transient channel functions) showed a marked deficit in the target location BM task. This suggests that the abnormality of subcortical transient channels does not explain some visual information processing dysfunctions in schizophrenia. Instead, deficient cortical interactions of rapidly changing environmental signals may be involved. PMID- 11065141 TI - Expression of inducible nitric oxide synthase mRNA and production of nitric oxide are induced by adenosine triphosphate in cultured rat microglia. AB - The effect of adenosine triphosphate (ATP) on the expression of the inducible nitric oxide synthase (iNOS) mRNA and production of nitric oxide in cultured rat microglia was examined. ATP induced iNOS mRNA dose-dependently (100-1,000 microM). The induction began within 1 h after addition of ATP (100 microM) with peak expression occurring at 6 h. The release of nitric oxide to culture medium was significantly increased by the treatment with ATP (100 and 1,000 microM) for 12 and 24 h. These results indicate that ATP is a potential mediator to induce iNOS mRNA expression and NO production in microglia. PMID- 11065142 TI - Confirmation of the genetic association of interleukin-1A with early onset sporadic Alzheimer's disease. AB - An association between altered risk of Alzheimer's disease (AD) and the interleukin-1A (IL-1A) gene was recently reported in two independent studies. Here, the IL-1A polymorphism was analyzed in a third population of 247 AD patients and 187 control individuals. In support of the published findings, the IL-1A 2/2 genotype was associated with an increased risk of early onset AD (odds ratio of 3.1, P = 0.02). Clinically, the IL-1A 2/2 genotype was associated with an earlier age of onset, but not with a change in the rate of progression of AD. PMID- 11065143 TI - Are P2Y1 purinoceptors expressed in turkey erythrocytes? AB - Since the beginning of purinoceptor research turkey erythrocytes have been widely used as the model systems for studying the pharmacology of P2Y1 nucleotide receptors. In this report the statistical analysis of the activity parameters of several purinoceptor agonists and antagonists in the turkey erythrocytes and P2Y1 receptor transfected cells is presented. As a results of this analysis several differences in the ligand activity orders measured in these biological systems were found. These data indicate that the receptors expressed in turkey erythrocytes and P2Y1 transfected cells are probably not the same. Whether it has to do with co-expression of several purinoceptor subtypes in turkey erythrocytes or novel P2Y receptors needs the further investigation. PMID- 11065144 TI - Detection of emerging resistance patterns within longitudinal surveillance systems: data sensitivity and microbial susceptibility. MYSTIC Advisory Board. Meropenem Yearly Susceptibility Test Information Collection. AB - Communicating information from antimicrobial resistance surveillance study data to microbiologists and physicians can be challenging. Large amounts of data, commonly reaching millions of MICs or zone diameter endpoints, must be analysed and condensed to easily-read tables or figures. Furthermore, data must not be prejudged relative to susceptibility categories, because of the diverse nature of interpretive criteria available internationally. An attempt must be made to present results of all surveillance studies in a mode that can be reinterpreted for immediate use in different geographical areas, or used to compare future data with relative ease and high accuracy. Such data displays require peer-reviewed journals to permit greater numbers of more complex tables to present results. The Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) study is a year-on-year global surveillance programme in medical centres where meropenem is available for use. We have developed a presentation strategy that expands the long-term clinical value of MYSTIC results. In addition to statistical parameters, tables of cumulative percentages or numbers of strains inhibited at each tested antimicrobial concentration will be presented. Alternative figures (Finland-o-grams) could also be used, but these generally lack precise extractable rates and require more journal space. Regardless of study design, promotion of this presentation philosophy enhances any surveillance study's value to each reader or user and facilitates application to locally appropriate interpretations. The widespread use of these analysis and presentation principles as benchmarks by various resistance studies and networks is strongly encouraged, particularly by investigations across international boundaries. PMID- 11065145 TI - MYSTIC (Meropenem Yearly Susceptibility Test Information Collection): a global overview. AB - The Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) is a global, multicentre surveillance study that compares the activity of meropenem in centres that are prescribers with that of imipenem, ceftazidime, piperacillin/tazobactam, ciprofloxacin and gentamicin. Of the 46 centres (intensive care units, cystic fibrosis units, neutropenia units and general wards) contributing to this study, 29 were in Europe, 14 in the Americas and three in the Middle East and Asia. The results for the most common isolates obtained in the first year of the study from these three regions show that meropenem has a broad spectrum of activity and potency in these centres, with 89% of the 6890 strains tested having an MIC < or = 4 mg/L. The overall susceptibility was lower for the comparator antibiotics. There was evidence in all regions of strains producing beta-lactamases and other resistance mechanisms against the other beta-lactams tested, fluoroquinolones and aminoglycosides. Future years' results from this surveillance study will show whether meropenem will continue to exhibit such activity. PMID- 11065146 TI - MYSTIC (Meropenem Yearly Susceptibility Test Information Collection) results from the Americas: resistance implications in the treatment of serious infections. MYSTIC Study Group (Americas). AB - The Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) programme aims to provide in vitro surveillance data for geographically diverse institutions where meropenem is available for use. The in vitro activity of meropenem and eight comparator antimicrobial agents against 2340 significant pathogens obtained in 1999 was assessed and compared in 14 study centres in Brazil, Mexico and the USA. Isolates were further characterized for production of extended-spectrum beta-lactamases (ESBLs), AmpC beta-lactamases and carbapenemases. Carbapenems demonstrated their broad spectrum and potency, inhibiting > or = 95% of all isolates irrespective of the geographical region or centre type. The overall order of activity of the nine agents tested against all pathogens in 1999 was meropenem (96%) > imipenem (95%) > cefepime (92%) > gentamicin (89%) > piperacillin/tazobactam (88%) > ceftazidime = tobramycin (86%) > cefotaxime (84%) > ciprofloxacin (83%). Thus far, the results from the Americas indicate that meropenem has excellent potency and spectrum of activity despite being prescribed for the treatment of seriously ill patients. In contrast, other ESBLs, fluoroquinolones and aminoglycosides have lost activity in many institutions as a result of the selection of strains producing ESBLs or having AmpC and other resistance determinants. Carbapenem resistance was observed rarely and at a prevalence similar to those reported in earlier studies. Carbapenems appear to be a continuing reliable option for the treatment of serious nosocomial infection. PMID- 11065147 TI - MYSTIC (Meropenem Yearly Susceptibility Test Information Collection) results from Europe: comparison of antibiotic susceptibilities between countries and centre types. MYSTIC Study Group (European centres only). AB - The activity of meropenem and five comparators has been studied against 7886 isolates from 29 centres in 10 European countries from 1997 to 1999 as part of the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC) surveillance study. Gram-positive and Gram-negative isolates from intensive care units (ICUs), neutropenia centres, cystic fibrosis (CF) centres and general wards were investigated in Belgium (1 year), Czech Republic (2 years), Germany (3 years), Italy (3 years), Poland (2 years), Russia (2 years), Sweden (2 years), Switzerland (1 year), Turkey (1 year) and UK (3 years). Resistance to quinolones and aminoglycosides was observed, as was resistance to the cephalosporins and penicillins via extended-spectrum beta-lactamases (ESBLs) and AmpC beta lactamases. Meropenem showed good activity against the pathogens tested, particularly in CF and neutropenia centres, over the 3 year period. The overall order of potency of the six antimicrobial agents tested was: meropenem > imipenem > piperacillin/tazobactam and ciprofloxacin > ceftazidime > gentamicin. No increase in resistance to the carbapenems, to date, has been detected in any of the European centres included in this study. PMID- 11065148 TI - Surveillance studies: how can they help the management of infection? AB - The increase in antimicrobial resistance has led to predictions of doom in the international press and to depression in the medical community. It has focused attention upon measures for fighting resistance, foremost of which is susceptibility surveillance. Until recently, global efforts at surveillance have been largely uncoordinated and random. This scene is rapidly changing with the World Health Organization (WHO), among others, leading multidisciplinary, targeted initiatives. In terms of individual surveillance programmes, much has been learned about their design. The best of these, the Meropenem Yearly Susceptibility Test Information Collection (MYSTIC), SENTRY and the Alexander Project, involve well-defined patient and organism groups against key denominators, and use standardized, internationally recognized methods that are quality-controlled, explore susceptibility quantitatively and include investigation of resistance mechanisms. Results are rapidly returned to the user. Evidence shows that surveillance, when used to guide policies on antibiotic use and infection control, can be helpful in the fight to control the development and spread of resistance. Further work is required to demonstrate these benefits and quantify them fully. PMID- 11065149 TI - Training for 'radionuclide radiology' in the UK. PMID- 11065150 TI - The role of positron emission tomography with fluorine-18-deoxyglucose in identifying colorectal cancer metastases to liver. AB - Liver metastasis is a common consequence of colorectal carcinoma. Early and accurate detection of liver metastasis is crucial for a decision about partial hepatectomy, which is considered a standard and potentially curative therapy in such a setting. The presence of extrahepatic metastases will exclude surgical resection as a therapeutic option. Positron emission tomography with fluorine-18 deoxyglucose (FDG-PET) has been successful in detecting and staging a variety of malignancies. The purpose of this study was to assess the utility of FDG-PET in the accurate detection of liver and distal metastases from colorectal cancer. The results of 80 PET and computed tomography (CT) scans were compared with surgical pathology and clinical outcome. FDG-PET detected liver metastases in 28 patients, with a sensitivity of 100%. CT detected metastasis in 20 patients, giving a sensitivity of 71.4%. In addition, in one patient with negative CT findings, PET detected a focus of hypermetabolism in the region adjacent to liver, which was proven to be a second focus of primary colon carcinoma. In six patients with liver metastases, PET correctly detected extrahepatic lesions, while CT only detected hepatic lesions. In conclusion, FDG-PET is an excellent imaging modality for the detection and staging of liver metastases in patients with colorectal carcinomas. PMID- 11065151 TI - Robotic synthesis of 6-[18F]fluoro-L-dopa. AB - The aim of this study was to develop a method of semi-automated 6-[18F]fluoro-L dopa (6-[18F]FDOPA) synthesis using a robotic system (Scanditronix Anatech RB III, Uppsala, Sweden). [18F]Fluorine was produced via 20Ne(d,alpha)18F using a Scanditronix MC17F cyclotron (Uppsala). The radiosynthesis was performed by the Scanditronix Anatech RB III robotic system. On average, a typical run produced 16 19 mCi of 6-[18F]FDOPA at end of synthesis (EOS) after 2 h irradiation of the F2/neon gas target. The total synthesis time was 110 min. The retention time of 6 [18F]FDOPA (the radio peak) was 8.2 min, which was consistent with the 6 [19F]FDOPA ultraviolet peak. The radiochemical purity was greater than 97%. A robotic, semi-automated method for 6-[18F]FDOPA radiosynthesis is therefore feasible. The radiation burden for the operator can be reduced as much as possible. Sufficient activity of 6-[18F]FDOPA could be obtained for positron emission tomography studies of dopaminergic function. PMID- 11065152 TI - Correlation of thallium-201 uptake with proliferating cell nuclear antigen in brain tumours. AB - The aim of this study was to assess the correlation between thallium-201 (201Tl) uptake and proliferative activity measured using the proliferating cell nuclear antigen labelling index (PCNA-LI) in brain tumours. Twenty-nine patients with brain tumours were included in this study. Of these, seven were diagnosed with meningioma, 13 had high grade glioma (HGG) and nine had low grade glioma (LGG). A 210Tl single photon emission computed tomography study was performed on all patients before operation, and 201Tl uptake index (UI) and retention index (RI) values were calculated. Cell proliferation was determined by PCNA. While all of the HGGs and meningiomas showed intense 201Tl accumulation on visual interpretation, eight of the nine LGGs did not show accumulation of 210Tl at the tumour site. 201Tl UI values were 3.23+/-0.89 and 2.67+/-0.66 in HGG, 1.27+/-0.18 and 1.23+/-0.09 in LGG, and 4.35+/-1.60 and 2.52+/-0.78 in meningioma on early and delay images, respectively. There was a statistical difference between the 201Tl UI in HGG and LGG. PCNA-LI values were 16.72+/-6.15%, 1.63+/-0.81% and 2.00+/-1.88% in HGG, LGG and meningioma, respectively. The PCNA-LI in HGG was significantly higher than in LGG and meningioma. While the correlation coefficient between the 201Tl UI and the PCNA-LI was 0.94 in gliomas (n=22), there was no correlation in meningiomas. No statistically significant correlation was found between the RI and the PCNA-LI in gliomas. The presence of a strong positive correlation between 201Tl uptake and PCNA-LI indicates that 201Tl uptake can predict the proliferative activity of the glioma. PMID- 11065153 TI - Improving the measurement of the 99Tc(m)-ECD brain perfusion index by temporal analysis. AB - Matsuda et al. have described a non-invasive method for brain perfusion quantification by computing the ratio of the cumulated counts in the cerebral hemispheres and aortic arch. The regions of interest (ROIs) are drawn manually and are observer dependent. The aim of this study was to develop a new method designed to minimize the intra- and interobserver variability when drawing the different ROIs. A dynamic study was performed as in Matsuda's method on 30 patients using technetium-99m ethyl cysteinate dimer (99Tc(m)-ECD) (ID: 800 MBq+/ 33 MBq). The manual method of drawing ROIs was then compared with the following, automated one. A temporal analysis was performed on the cardiac first-pass study to obtain parametric images of the thorax. An ROI of the aortic arch was drawn automatically by means of an isocontour algorithm on the resulting views. The whole sequence was reframed and filtered by a temporal low-pass filter. Hemispheric brain ROIs were delineated on a summed image. Matsuda's algorithm was then applied. Intraobserver variability was evaluated for the classical Matsuda method. The correlation in brain perfusion index (BPI) measurements was r = 0.8976 for naive observers and r = 0.9443 for well-trained observers. Interobserver variability was also evaluated; the correlation was r = 0.7574 for naive observers and r = 0.9190 for well-trained observers. With our proposed method, the correlation in the measurements of BPI for evaluating the intraobserver variability was r = 0.9955 for naive observers and r = 0.9989 for well-trained observers. For interobserver variability, the results were r = 0.9234 for naive observers and r = 0.9230 for well-trained observers. We conclude that temporal analysis allows brain perfusion to be measured in a semi-automatic manner, and improves the reproducibility compared with the original method of Matsuda, particularly for naive observers. PMID- 11065154 TI - Investigation of perfusion reserve using 99Tc(m)-MIBI in the lower limbs of diabetic patients. AB - This study aimed to investigate the microvascular pathology in the lower limbs of diabetic patients without symptoms or findings of peripheral ischaemia by measuring perfusion reserve scintigraphically. It was carried out in 47 female subjects who had no evidence of peripheral arterial disease in their history, physical examination or Doppler ultrasonography. The diabetic group consisted of 25 women (mean age 54.2 +/- 3.54 years) with type II diabetes mellitus of more than 10 years' duration. A control group consisted of 22 healthy non-diabetic women (mean age 50.14 +/- 6.75 years). Each subject flexed their right foot maximally both dorsally and plantar 60 times. In the middle of this exercise, 370 MBq technetium-99m-methoxyisobutylisonitrile (99Tc(m)-MIBI) was injected intravenously. Ten minutes after the injection, a posterior image of both calves was obtained using a gamma camera. Rectangular regions of interest were symmetrically drawn over both calves. The total count in the resting calf was subtracted from the total count in the exercising calf, and the percentage increase, termed the perfusion reserve, was determined. A significant difference was found between the perfusion reserves of the diabetic and control groups (76.04 +/- 12.96% and 95.91 +/- 12.83%, respectively; P<0.001). In conclusion, microvascular pathology may be determined scintigraphically by measuring the perfusion reserve in the lower limb muscles in diabetic patients. This method may also be used to evaluate perfusion abnormalities in other circulatory disorders. PMID- 11065155 TI - Optimum filtration for time-activity curves in nuclear medicine. AB - Insufficient filtration and over-smoothing are misleading processes in the quantification of time-activity curves. The optimum filtration requires a good knowledge of the frequency spectrum and relative amplitudes of the data and superimposed noise. Due to variations in biomedical data, it is very difficult to adjust the filter for individual cases. To overcome this problem a new method of noise reduction is proposed. In this method the time-activity curves are transformed into a low frequency (linear) curve that can be filtered heavily without significant distortion of the real data. The theory of the proposed filter and the results of its comparison with three-point filter, five-point filter and data bounding methods are presented. The comparison was performed using deconvolution analyses of simulated renograms. The results show that the proposed filter causes minimum distortion of the renogram and impulse retention function in terms of the root mean square error and the peak of the renogram. Moreover, the filter is much less sensitive to over-smoothing (number of filter iterations), the signal-to-noise ratio and the mean transit time of the renogram compared with other filters. PMID- 11065156 TI - An audit of biliary scintigraphy in a district general hospital (1993-1998) with special reference to the investigation of acalculous gallbladder disease. AB - One hundred and ninety nine cholescintigrams were performed at our hospital between January 1993 and December 1998. Of these, 96 were performed as part of the investigation of right upper quadrant or epigastric pain in patients whose prior biliary ultrasound scans had shown no abnormalities. In this group of patients, 60 cholescintigrams were reported as normal, 28 as abnormal and eight as equivocal. A review of the case notes for this group was made, with 91 out of 96 case notes being retrieved. Thirteen of the 58 (22%) patients with normal cholescintigrams either underwent or had been offered cholecystectomy, compared with 22 of the 27 (81%) with 'definitely abnormal' cholescintigrams. Of the subset of patients with abnormal cholescintigrams subsequently undergoing cholecystectomy, 16 out of 18 were found to have histologically abnormal gallbladders, while 11 out of 18 reportedly had their symptoms cured or improved. A fatty meal is used in our unit as the stimulus to gallbladder contraction, in contrast to the majority of papers published in the literature. We believe that the use of a fatty meal is acceptable and that the use of intravenous exogenous cholecystokinin is probably best reserved for the small group of patients with equivocal scintigrams who may require further investigation. PMID- 11065157 TI - Effects of octreotide and morphine on the clearance rate of indium-111 pentetreotide from the epidural space. AB - In this study we aimed to evaluate the possible mechanisms by which somatostatin acts when given epidurally. Twenty male New Zealand rabbits were randomly separated into four groups and various drugs were administered via a caudal epidural catheter. Group 1 received a bolus of 3.7 MBq indium-111 ((111)In) pentetreotide, group 2 received 200 microg octreotide and after 15 min a bolus of 3.7 MBq (111)In-pentetreotide, group 3 received 0.1 mg morphine and after 15 min a bolus of 3.7 MBq (111)In-pentetreotide, and group 4 received a bolus of 3.7 MBq technetium-99m (99Tc(m))-diethylene triamine pentaacetic acid (DTPA). Dynamic images of 60 min' duration were obtained from the posterior projection. T(1/2), fast and T(1/2) total clearance half-times were calculated. When unlabelled octreotide was given to block somatostatin receptors, clearance of (111)In pentetreotide was found to be faster. Epidural morphine administration did not change the clearance rate of (111)In-pentetreotide. All these findings are in favour of octreotide binding to its probable own specific receptors present in the epidural space. PMID- 11065158 TI - Spleen-targeted 114In(m) via normal and heat-damaged red cells in the mouse: isotope distribution and bone marrow damage. AB - 114In(m)-labelled heat-damaged erythrocytes (HDRBC) in BDF1 mice were quantitatively much less successful in spleen targeting than in the rat. Unheated labelled cells (NRBC) were an even less effective vector. Radiobiological effects manifest in the spleen (weight loss) and white cell count were nevertheless substantial, comparable in both groups and unchanged after 14 days. Considerable early renal loss of indium is inferred in the NRBC animals, but in the HDRBC group a high proportion was retained in the liver. Progenitor cell marrow cultures indicated acute transitory lethal effects with only partial recovery at termination; the marrow concentration of indium was the same in both groups. PMID- 11065159 TI - Technetium-99m-tetrofosmin: retention of nitrogen atmosphere in kit vial as a cause of poor quality material. AB - Technetium-99m (99Tc(m))-tetrofosmin was prepared using four different reconstitution methods. The radiochemical purity (RCP) of these products was assessed 8 h later using thin layer chromatography (TLC). Material produced using the original method supplied by the manufacturer and using an newer method, which involves the use of a vent needle and the addition of air, had acceptable RCP (mean +/- SD 94.2 +/- 1.1% and 94.7 +/- 1.7%, respectively) and similar chromatograms. In addition, both products showed good clinical efficacy and exhibited normal biodistribution behaviour. Preparing 99Tc(m)-tetrofosmin using the two other methods, one using a high radioactive concentration and the other maintaining the nitrogen content of the kit vial, gave rise to chromatograms with reduced RCP (63.5 +/- 10.9% and 61.9 +/- 7.6%, respectively) and greater levels of impurities. Although neither of these last two preparations was used clinically, we suggest that reports of poor quality images may be the result of administration of materials similar to these. Results for the high radioactive concentration method were as expected and are consistent with the restrictions imposed by the manufacturer. However, results using the last method are surprising and would suggest that the production of good quality 99Tc(m) tetrofosmin is dependent on the quantity of nitrogen in the kit vial. We believe that the amount of nitrogen removed from the kit vial during the process of reconstitution is critical. If too much nitrogen is present this will result in poor quality material. In practice it is conceivable that there could be occasions when insufficient nitrogen is removed when following the manufacturer's original guidance, thereby leading to low RCP material. To ensure adequate nitrogen is removed during reconstitution, adoption of the manufacturer's revised method, involving the deliberate introduction of air, is therefore appropriate. PMID- 11065160 TI - The structural failure of a multi-hole collimator. AB - During measurements of the extrinsic spatial resolution of a gamma camera, it was noted that there was a variation with the position in the field of view. The problem was traced to the collimator, and this was confirmed by inspection of the parallel hole structure. Removal of the cover showed that the lead foil honeycomb had become detached from its surround. The fault could have had serious implications for the quality of patient investigations and could also have caused damage to the crystal if it had gone undetected. As a result of this investigation, the quality assurance procedures within this department have been updated to include a visual check of all collimator cores. PMID- 11065161 TI - Direct deconvolution algorithms based on Laplace transforms in nuclear medicine applications. AB - A set of algorithms is presented for direct deconvolution of the residue signal of an organ with the input signal to the organ. The deconvolution process yields the residual impulse response from which the distribution of transit times and the important mean transit time can be readily determined. The deconvolution method is based on the Laplace transform and it requires that the input signal can be fitted with an expression consisting of one, two or three exponentials with or without a bolus term at zero time or a constant term. These types of exponential expressions for the input signal cover a wide range of the input signals encountered in nuclear medicine applications. Simulation studies of the residue signal by convolution of various input signals with a number of residual impulse response models yielded an excellent accuracy of the deconvoluted residual impulse response for a suitably small sampling time. The simulations provide an opportunity to understand further the shapes of the residue curves depending on the shape of the input signal and the distribution of transit times. Simulations with Gaussian-distributed noise and noise spikes superimposed on the residue signal were also made to investigate the robustness of the direct deconvolution algorithm using apparently real-life data. PMID- 11065162 TI - 99Tc(m)-MAG3: problems with radiochemical purity. PMID- 11065163 TI - Justification and controllable dose. PMID- 11065164 TI - Troglitazone and related compounds: therapeutic potential beyond diabetes. AB - Troglitazone and structurally related compounds (pioglitazone, rosiglitazone etc.) containing thiazolidinediones (TZD) are a novel class of antidiabetic agents which decrease blood glucose in diabetic animal models and in patients with Non-Insulin-Dependent Diabetes Mellitus (NIDDM) through alleviating insulin resistance. A large body of evidence is now accumulating indicating that insulin resistance and/or resulting hyperinsulinemia underlie the pathogenesis of not only diabetes but also of the clustering syndrome called "syndrome X" or "insulin resistance syndrome" which includes hypertension, dislipidemia and hypercoagulation. Therefore, TZD class of insulin sensitizers seem to have therapeutic potential to improve this clustering syndrome in addition to diabetes. Moreover, it was demonstrated that the TZD class of insulin sensitizers including troglitazone bind and activate the peroxisome proliferator-activated receptor gamma (PPARgamma), a nuclear hormone receptor. Although PPARgamma is predominantly expressed in adipose tissue, one of the target tissues for insulin, it have been subsequently found to be expressed in macrophages, vascular smooth muscle cells (VSMC), endothelial cells and several cancer cell lines. PPARgamma activation by PPARgamma agonists such as TZD class of insulin sensitizers in these cells modulates these cell functions such as the production of inflammatory cytokine by macrophages, proliferation and migration of VSMC, and growth or differentiation in cancer cells. In addition, troglitazone has potent antioxidant effect, and suppresses both L-type and receptor operated Ca2+ channel and protein kinase C. Thus since TZD class of insulin sensitizers has many kind of therapeutic effect in addition to lowering blood glucose, these agents expect to have therapeutic potential beyond diabetes. PMID- 11065165 TI - Liver gene expression and increase in albumin synthesis by fetal hepatocytes transplanted into analbuminemic rats. AB - This report describes the evolution of hepatocytes isolated from 21-day fetuses and transplanted into spleens of Nagase analbuminemic rats which have negligible serum albumin levels due to a mutation affecting albumin mRNA processing. Albumin and alpha-fetoprotein expression, in addition to other parameters related to cellular proliferation status (thymidine kinase and proliferating cell nuclear antigen expression) were studied as indicative of the behavior and evolution of the cells. In recipient rats, only a few clusters of hepatocytes could be observed in the red pulp of the spleen 24 h after transplantation. The fetal hepatocytes migrated to the liver and could be seen in portal branches immediately after transplantation. Fifteen days later, albumin mRNA was detected in recipient livers and was expressed throughout the entire 3-month study. Alpha fetoprotein was not detected. Cell proliferation was not relevant, although 3 months after transplantation, the proliferation rates appeared to show a tendency to increase. These data demonstrate that fetal hepatocytes transplanted into spleen migrate to liver, settle there and acquire an adult phenotype free of malignant transformation. Our study is a first step towards the thorough understanding of fetal hepatocyte transplantation. The next steps will involve in depth studies of the possibilities of genetic manipulation to achieve a high degree of repopulation/expression, employing the least possible number of donor cells, and of how the cells reach the liver parenchyma, overcoming the endothelial barrier. PMID- 11065166 TI - Elevated serum leptin concentrations induced by experimental acute inflammation. AB - Leptin is a pleiotropic hormone that regulates body weight and energy expenditure. Recent findings suggest that leptin may be involved in acute and/or chronic inflammation, however only limited results are available describing the effects of in vivo models of acute inflammation on leptin secretion. The aim of this study was to evaluate serum leptin levels in response to two well established models of acute inflammation in rats: carrageenan rat paw induced oedema and carrageenan induced pleurisy. Our results clearly show that leptin levels rise in rats in which both oedema and pleurisy were induced. Serum leptin levels in carrageenan induced paw oedema were 3.86+/-0.16 microg/L in comparison to 1.83+/-0.17 microg/L of control animals (p<0.001). A similar result was observed in carrageenan induced pleurisy animals in which leptin levels were 4.87+/-0.27 microg/L in comparison to 2.19+/-0.16 microg/L of control animals (p<0.001). The increase in leptin levels induced following carrageenan-induced pleurisy appears to be dependent on adrenal function and it is markedly blunted in adrenalectomized rats. Leptin levels in carrageenan induced pleurisy, carried out on adrenalectomized rats, were lower than intact inflamed animals, suggesting a possible involvement of endogenous glucocorticoids. In summary the results here presented show that: a) an elevated plasma leptin concentration was induced during experimental models of inflammation b) this increase is mediated to a large extent by glucocorticoids. In conclusion, acute experimental models of inflammation are associated with changes in circulating leptin suggesting a possible involvement of this hormone in the anorexia/cachexia that is frequently associated with inflammatory processes. Furthermore, our data indicate the existence of a feedback loop among glucocorticoids and leptin which might contribute to the immune response to lace the inflammatory process. PMID- 11065167 TI - Pharmacological characterization of [3H]-JTH-601, a novel alpha1-adrenoceptor antagonist: binding to recombinant human alpha1-adrenoceptors and human prostates. AB - Several alpha1-adrenoceptor (AR) selective antagonists are now widely used to improve lower urinary tract symptoms in benign prostatic hyperplasia patients. However, these drugs often result in orthostatic hypotension, because of their poor uroselectivity; the blockade of alpha1-AR not only in prostate but also in vasculature. Here we have investigated uroselectivity of JTH-601, a newly developed antagonist, in radioligand binding experiment using recombinant human alpha1-AR subtypes and human prostate. In saturation experiments, [3H]-JTH-601 showed subtype selectivity: high affinity to alpha1a-AR (pKd; 9.88+/-0.09), lower affinity to alpha1b-AR (pKd; 8.96+/-0.17) and no specific binding at concentrations up to 3000 pM to alpha1d-AR. In competition experiments, JTH-601 and its metabolic compound (JTH-601-G1) also showed alpha1a-AR selectivity, exhibiting approximately 5 times higher affinity for alpha1a-AR than for alpha1b AR, 10 to 20 times higher affinity than for alpha1d-AR, respectively. [3H]-JTH 601 also bound to human prostate membranes in monophasic manner with high affinity constant (pKd; 9.89+/-0.12, Bmax=123.6+/-16 fmol/mg protein). JTH-601 is a unique alpha1-AR antagonist that shows high affinity and selectivity for human recombinant alpha1a- and human prostate. This new compound is useful for understanding alpha1-AR pharmacology and may have a therapeutic value. PMID- 11065168 TI - Characteristic glucuronidation pattern of physiologic concentration of morphine in rat brain. AB - Formation of conjugated metabolites from morphine at a very low level in brain was studied in vitro in rats. Incubation of a low concentration of 3H-morphine with brain homogenate followed by two successive high-performance liquid chromatographic analyses showed that endogenous morphine is converted by brain enzymes to its 3- and 6-glucuronides (M-3-G and M-6-G), and codeine glucuronide (Cod-G). However, the formation of morphine-6-sulfate was likely to be low if it was produced at all. All of the cerebral hemisphere, brain stem and cerebellum were capable of producing M-3-G, M-6-G and Cod-G, although there were differences in selectivity. The capacity of the brain for glucuronide formation was far less than that of the liver, but UDP-glucuronosyltransferase in brain was much more selective in forming M-6-G and Cod-G than liver enzymes. PMID- 11065169 TI - Effects of intermittent hypoxia on action potential and contraction in non ischemic and ischemic rat papillary muscle. AB - Although it has been reported that intermittent hypoxia had the anti-arrhythmia effect, little is known about the effects on the action potential (AP) and contraction of papillary muscle, as well as the mechanism of anti-arrhythmia. The purpose of present study is to observe the effects of intermittent hypoxia on action potential and contraction of papillary muscle in rat left ventricle simultaneously using conventional intracellular microelectrode and contraction recording. The effects of intermittent hypoxia on AP and contraction during ischemic solution perfusion were also investigated. After exposed to intermittent hypoxia (six hours daily) for 42 days (IH42), duration (APD20) of 20%, 50% (APD50) and 90% (APD90) repolarization of AP prolonged significantly compared with animals in control (Con). Effective refractory period (ERP) in IH42 also prolonged significantly. Perfused with mimic ischemic solution, the changes of electric and mechanical activities in IH42 and in 28 days exposure to intermittent hypoxia (IH28) were much smaller than that in Con and IH14. The result of the study suggested that intermittent hypoxia prolonged the APD and ERP, offered the resistance against the ischemic damage on myocardium, which may be the electrophysiological basis of the anti-arrhythmia of intermittent hypoxia. PMID- 11065170 TI - Production of human UDP-glucuronosyltransferases 1A6 and 1A9 using the Semliki Forest virus expression system. AB - Human UDP-glucuronosyltransferases (UGTs) 1A6 and 1A9 were expressed using Semliki Forest virus (SFV) vectors. Infection of chinese hamster lung fibroblast V79 cells with recombinant SFV-UGT viruses resulted in efficient protein expression as detected by metabolic labeling, Western blot analyses and immunofluorescence microscopy. The expression of UGT 1A6 and UGT1A9 in the SFV infected cells was approximately two fold higher than in a stable V79 cell line. No UGT signal was detected in noninfected cells. In addition, SFV-UGT viruses also efficiently infected other mammalian cells, such as baby hamster kidney (BHK), chinese hamster ovary (CHO) and human lung (WI-26 VA4) cells leading to high production of recombinant enzyme. The measurement of enzyme activities and kinetic parameters using p-nitrophenol and nitrocatechol (entacapone) as substrates for UGT1A6 and UGT1A9, respectively, showed that the overall kinetic properties of the enzymes produced by the two systems were similar. We conclude that the SFV expression system represents an efficient, fast and versatile method for production of metabolic enzymes for in vitro assays. PMID- 11065171 TI - T-butyl hydrogen peroxide increases the activities of the Maxi-K channels of rat brain. AB - In this study, we investigated the effects of tertiary-butyl hydrogen peroxide (tBHP) on the large-conductance Ca2+-activated K+ (Maxi-K) channel of rat brain using lipid bilayer. When tBHP was applied to the cytosolic side, the open probability (Po) of both fast- and slow-gating Maxi-K channels increased within 1 min in dose-dependent manner. tBHP effects did not reverse immediately, suggesting tBHP induces some chemical modification on the channel protein. From kinetic analysis of single channel data, the increase in the Po appears to be mainly due to shortening of closed dwell time in both types of the Maxi-K channels. 50 microM diamide, a sulfhydryl-specific oxidant, irreversibly decreased the Po. However, further addition of 7.3 mM tBHP still increased the Po, suggesting that tBHP does not share the target for oxidation with diamide. PMID- 11065172 TI - Early effects of exogenous arginine after the implantation of prosthetic material into the rat abdominal wall. AB - We have investigated the effects of high arginine (Arg) levels (7.5 mg/100 g body weight per hour) on the early integration of biocompatible mesh grafts into the rat abdominal wall. Studies were performed over implantation intervals of 6, 12, 24 or 48 hours (n=12, each). Arginine and related compounds were quantified in plasma, wound fluids and multiple tissues. Plasma nitric oxide (NO) production was studied. Strips were taken from the polypropylene fiber-host tissue interfaces (PTIs) for optical microscopic analysis and for immunohistochemical analysis using rat-specific antibodies against type I and type III collagens. Exogenous Arg was metabolized at the peripheral tissues but reliably reached the wound space. High amounts of Arg and ornithine (Orn) were detected in the specimens considered. No changes on citrulline (Ctr) or NO concentrations were observed, overall suggesting that, during the period studied, the arginase pathway predominated. The acute scarring response differed significantly in the two placements considered. The P-SS interface evidenced more extensive new tissue growth than the P-DS interface. Forty-eight hours after mesh implantation cellular infiltration, fibroblast proliferation, and mesh-surrounding angiogenesis were higher in the arginine-treated rats. Type III collagen staining was related to arginine treatment, being higher (++) in the study group. In conclusion, and independently of the site of mesh placement, supplemental Arg seemed to favorably affect early local collagen deposition. This could be potentially helpful to ameliorate the integration of biomaterials into the tissues and, consequently, to allow for the design of more selective therapeutic strategies to prevent hernia recurrence rates. PMID- 11065173 TI - Pimobendan inhibits the activation of transcription factor NF-kappaB: a mechanism which explains its inhibition of cytokine production and inducible nitric oxide synthase. AB - Pimobendan, an inhibitor of phosphodiesterase III with calcium sensitizing properties, inhibits the production of cytokines and nitric oxide. In the present study, the effects of pimobendan and other inotropic agents on the activation of transcription factor NF-kappaB were examined. Pimobendan significantly decreased the expression of luciferase protein in A549 cells transfected with the NF-kappaB reporter plasmid, stimulated with interleukin (IL)-1beta, tumor necrosis factor alpha, or phorbol 12-myristate 13 acetate. However, high concentrations of amrinone, vesnarinone, or NKH 477 decreased promoter activity only slightly. Electrophoretic mobility shift assay also showed inhibition of NF-kappaB activation by pimobendan. Pimobendan possesses the unique property of inhibiting NF-kappaB, which may be independent of phosphodiesterase inhibition. PMID- 11065174 TI - Characterization of the protease-activated receptor-1-mediated contraction and relaxation in the rat duodenal smooth muscle. AB - Activation of protease-activated receptor-1 (PAR-1) produces a dual action, apamin-sensitive relaxation followed by contraction, in the rat duodenal smooth muscle, which is partially dependent on activation of L-type Ca2+ channels, protein kinase C (PKC) or tyrosine kinase (TK), and resistant to tetrodotoxin. The present study further characterized the PAR-1-mediated duodenal responses. Removal of extracellular Ca2+ as well as SK&F96365 reduced the contraction due to the PAR-1 agonist TFLLR-NH2 (TFp-NH2) by 60-80% that was similar to the extent of the inhibition by nifedipine. Lowering of the extracellular Na+ concentration, but not IAA-94, a Cl- channel inhibitor, reduced both the PAR-1-mediated contraction and relaxation by about 50%. U73122, a phospholipase C (PLC) inhibitor, or wortmannin, a phosphatidyl inositol 3'-kinase (PI3K) inhibitor, significantly reduced the PAR-1-mediated contraction, but not the relaxation, by itself, as the PKC inhibitor GF109203X and the TK inhibitor genistein did. U73122 or wortmannin, like GF109203X, when applied in combination with genistein, significantly reduced the PAR-1-mediated relaxation. The relaxation was resistant to antagonists of PACAP receptors, VIP receptors and P2 purinoceptors. Thus, the PAR-1-mediated contraction is considered to be dependent on intracellular and extracellular Ca2+, the influx of the latter being induced through activation of L-type Ca2+ channels triggered by the enhanced Na+ permeability, and that PLC and PI3K, in addition to PKC and TK, are involved in the PAR-1-mediated dual responses. Furthermore, non-adrenergic, non-cholinergic nerve neurotransmitter candidates that may modulate K+ channels do not appear to contribute to the relaxation by PAR-1 activation. PMID- 11065175 TI - Effect of neonatal treatment with mifepristone or tamoxifen on the binding capacity of the thymic glucocorticoid or uterine estrogen receptor of adult rats: data on the mechanism of hormonal imprinting. AB - For studying the mechanism of perinatal hormonal imprinting newborn rats were treated with a single injection of the antihormones, mifepristone (RU486) or tamoxifen (100 microg each). Glucocorticoid receptors of thymi of 6 weeks old male and female, and uterine estrogen receptors of 2 months old female rats were studied for dexamethasone or estradiol binding, respectively. Tamoxifen caused faulty imprinting both in the thymic and uterine receptors, increasing affinity and density of males, and decreasing females' glucocorticoid receptors as well, as decreasing the density of uterine estradiol receptors. Neonatal mifepristone treatment was indifferent to the thymus, and decreasing to density of uterine estrogen receptors. Males' body weight significantly decreased 6 weeks after tamoxifen treatment. The results suggest that imprinting can not be provoked by a molecule (hormone antagonist) which can bind to the receptor without any postreceptorial events (mifepristone/glucocorticoid receptor), in the presence of some postreceptorial effects the reaction takes place, however the strongest reaction can be observed by the hormone analogue (tamoxifen) with postreceptorial (agonist) effect, not considering that the receptor is the direct target of the molecule or a cross-reaction is present. PMID- 11065176 TI - Change of the involvement of 5-HT3 receptor in the gastric motor stimulating actions of KW-5139 (Leu13-motilin acetate) in the recovered and post-operative periods in dogs. AB - We have previously reported that KW-5139, a motilin analogue, evokes gastrointestinal motor stimulating action in the post-operative period as well as in the recovered period of conscious dogs. In this report, we compared the mechanisms of the KW-5139-induced contractions in the post-operative period with those in the recovered period using beagle dogs implanted force transducers in the gastric antrum, duodenum, jejunum, ileum and colon. In addition, we also examined the mechanisms of the prostaglandin F2alpha-induced contractions in both periods. The gastric contractions evoked by KW-5139 (0.5 microg kg(-1), i.v.) were inhibited by the pretreatment of ondansetron (0.1 mg kg(-1), i.v.), a 5-HT3 receptor antagonist, in the recovered period, but were not affected in the post operative period even by higher doses of ondansetron (0.3-1 mg kg(-1), i.v.). The KW-5139-induced contractions in the small and large intestine were not inhibited by ondansetron in the both periods. The contractions evoked by KW-5139 (0.5 microg kg(-1), i.v.) in the gastric antrum, duodenum, jejunum and colon were significantly inhibited by the pretreatment with atropine (0.05 mg kg(-1), i.v.), a muscarinic receptor antagonist, in the recovered period as same extent as in the post-operative period. The contractions evoked by prostaglandin F2alpha (50 microg kg(-1), i.v.) in the any recording sites were not affected by the pretreatment with ondansetron (0.1 mg kg(-1), i.v.) in the recovered period. On the other hand, atropine (0.05 mg kg(-1), i.v.) tended to inhibit the gastric and colonic contractions. These effects of ondansetron and atropine on the prostaglandin F2alpha-induced contractions were not different between in the post operative and recovered periods. The present results indicate that 5-HT3 receptors are involved in the KW-5139-induced motor stimulating action in the recovered period but not in the post-operative period. The mechanisms of the alteration were discussed. PMID- 11065177 TI - Action of chronic CC14 on the retinol and dolichol content of rat liver parenchymal and non-parenchymal cells. AB - We studied dolichol, on account of its role in membrane fluidity and fusion, and retinol, on account of its behaviour in liver fibrosis, in isolated parenchymal and sinusoidal rat liver cells after CCl4 treatment for 3, 5 and 7 weeks. Retinol uptake was also investigated by administering a load of retinol three days before sacrifice. In hepatocytes, dolichol decreased and seemed to be the preferred target of lipid peroxidation by CCl4; indeed, retinol increased especially after vitamin A load. Two subfractions of hepatic stellate cells were obtained: in the subfraction called Ito-1, dolichol decreased, while the supplemented retinol was no longer stored; in the subfraction called Ito-2, the values were intermediate. In Kupffer and endothelial cells dolichol was higher after three weeks, in agreement with fibrogenesis. Retinol increased after retinol load, in Kupffer and endothelial cells, in agreement with their scavenger function. The different behaviour of dolichol content in parenchymal and non-parenchymal cells suggests that dolichol may have different functions in liver cells. Since it has been ascertained that, in liver fibrosis, stellate cells gradually lose retinol, the inability of HCs to send retinol to Ito-1 subfraction or the inability of Ito-1 subfraction to take up and store vitamin A might induce or contribute to the transformation of these cells into a different phenotype. This behaviour is discussed regarding the role of cellular and retinol binding proteins in intracellular retinol content. Moreover a role of dolichol in membrane fluidity and retinol traffic is hypothesised. PMID- 11065178 TI - Changes in GIRK1/GIRK2 deactivation kinetics and basal activity in the presence and absence of RGS4. AB - The effect of RGS4, a GTPase-activating protein, on the deactivation kinetics and basal activity of GIRK1/GIRK2 channels activated by the human kappa-opioid receptor (hKOR) was investigated. Co-expression in Xenopus oocytes of RGS4 reduces the basal GIRK1/GIRK2 current and strongly increases the percentage agonist-evoked K+ conductance. RGS4 reconstitutes the native gating kinetics by accelerating GIRK1/GIRK2 channel deactivation, a phenomenon also seen after activation with other 7 TM receptors (e.g. muscarine type). In the absence of RGS4, the GIRK1/GIRK2 conductance was increased by approx. 50% after hKOR stimulation with the kappa-selective opioid receptor ligand, U69593; however more importantly, at the end of the washout period it was dramatically reduced to about 60% of the basal conductance as measured before receptor stimulation. Furthermore, we found that repeated receptor stimulation causes an increase of the agonist-gated deactivation kinetics, without affecting the maximal and minimal conductance levels of GIRK1/GIRK2 channels during and after agonist application. Unlike in the absence of RGS4, coexpression with RGS4 completely abolished the reduction of basal conductance after agonist washout and the deactivation kinetics remained unaffected upon repeated agonist application. The results presented here clearly indicate that previous stimulation by agonists activating G protein-coupled receptors may have long-lasting, strong consequences on the following responses. Therefore, our study provides evidence for a novel modulation of deactivation kinetics of GIRK1/GIRK2 currents in the absence of RGS4. PMID- 11065179 TI - Effects of different serotypes of Escherichia coli lipopolysaccharides on body temperature in rats. AB - The effects of Escherichia coli O55:B5, O127:B8, and O111:B4 serotypes' lipopolysaccharides (LPS) on body temperature were investigated in rats. LPSs were injected intraperitoneally at doses of 2, 50, and 250 microg/kg. A multiphasic and no-dose dependent increase in rectal temperature was observed in response to E. coli O55:B5 LPS at all doses, and in response to E. coli O127:B8 LPS at 2 and 50 microg/kg doses. The highest dose of the latter caused a dual change in rectal temperature, in which hypothermia preceded fever. E. coli O111:B4 LPS was either pyrogenic or hypothermic at 2 and 250 microg/kg doses; respectively, whereas a dual response was observed when the 50 microg/kg dose was injected. Although dual responses were observed after administration of all LPSs at 50 microg/kg dose when the body temperature was recorded by biotelemetry, the hypothermia induced by E. coli O55:B5 LPS was significantly smaller. These data suggest that LPSs induce dose and serotype-specific variable changes on body temperature in rats. This variability may be related to the structure of LPSs. The data also indicate that LPS causes hypothermia with or without fever in rats. PMID- 11065180 TI - Involvement of Na+ and Ca2+ channel activation and resultant nitric oxide synthesis in glutamate-mediated hypoxic injury in rat cerebrocortical slices. AB - The role of Na+ and Ca2+ channels in glutamate-mediated hypoxic injury was investigated in slices of the rat cerebral cortex. Hypoxic injury was determined by mitochondrial reduction of 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyltetrazolium bromide after exposure of brain slices to 30-min of hypoxia/glucose deprivation followed by 3-h of reoxygenation. Endogenous glutamate release was markedly elevated during hypoxia/glucose deprivation, but it returned almost to basal level during reoxygenation. Hypoxic injury was prevented by MK-801 or 6-cyano-7 nitroquinoxaline-2,3-dione. Combined treatment with omega-conotoxin GVIA, omega agatoxin IVA, and tetrodotoxin reversed the hypoxic injury, although none of these agents alone or nifedipine was effective. Moreover, a novel Na+/Ca2+ channel blocker NS-7 [4-(4-fluorophenyl)-2-methyl-6-(5-piperidinopentyloxy) pyrimidine hydrochloride] significantly inhibited the hypoxic injury. Several inhibitors of nitric oxide synthase also blocked the hypoxic injury. Consistently, nitric oxide synthesis, as estimated from cyclic GMP formation in the extracellular fluids, was enhanced during hypoxia/glucose deprivation. NS-7 and other Na+ and Ca2+ channel blockers suppressed the enhancement of nitric oxide synthesis, although these compounds alone, or in combination, did not reduce hypoxic glutamate release. These findings suggest that hypoxic injury in rat cerebrocortical slices is triggered by glutamate and subsequent enhancement of nitric oxide synthesis through activation of both Na+ and Ca2+ channels. Thus, the simultaneous blockade of both Na+ channel as well as N-type and P/Q-type Ca2+ channels is required to sufficiently reverse the hypoxic injury. PMID- 11065181 TI - Altering expression of alpha3beta1 integrin on podocytes of human and rats with diabetes. AB - The adhesion molecule integrin alpha3beta1 is the major receptor of podocyte to the glomerular capillary basement membrane (GBM). Since progressive alteration of the glomerular extracellular matrix (ECM) compartment leading to GBM thickening is common in diabetic nephropathy, we investigated the cellular distribution of alpha3beta1 integrin in podocytes of patients with diabetic nephropathy and streptozotocin-induced diabetic rats, and we evaluated the effects of high glucose on the cultured rat podocytes. Both human and rat kidneys were stained using the immunoelectron microscopy and immunoperoxidase technique with mouse monoclonal antibodies to human integrin alpha3 subunit. The results showed that both the number of immunogold particles and the staining of integrin alpha3 subunit on podocytes were weaker in patients with diabetic nephropathy than those of control kidneys. The staining of alpha3 on podocytes in the poorly-controlled diabetic rats was also weaker after one and three months of hyperglycemia. However, the staining was identical to controls in rats with only one week of hyperglycemia. High glucose (25 mM) but not streptozotocin in vitro suppressed the alpha3 expression of cultured rat podocytes. Our results demonstrated that the expression of integrin alpha3beta1 on podocytes was suppressed in both human and rats with diabetes, possibly due to the effects of hyperglycemia, and the suppression became more severe with the duration of diabetes. PMID- 11065182 TI - Myelination deficits in brain of rats following perinatal asphyxia. AB - Perinatal asphyxia remains a major cause of acute mortality and of permanent neurodevelopmental disability in infants and children. However, the pathophysiologic features of hypoxic-ischemic encephalopathy are still incompletely understood. Animal studies have been focussing on grey matter pathology but information on white matter lesions is limited. The aim of the study was to investigate white matter lesions after three months following graded perinatal asphyxia in the rat using a well-documented, reproducible, clinically relevant and simple animal model of perinatal asphyxia. Brains of rat pups (n=10 per group) exposed to asphyctic periods of 10 and 20 minutes were examined histologically and compared to normoxic brain using Kluever-Barrera myelin staining, immunohistochemically with antibodies against myelin basic protein, 2',3'-cyclic-nucleotide'-phosphodiesterase as markers for myelination, antibodies against neurofilaments for the evaluation of axonal density and antibodies against glial fibrillary acidic protein as a marker for astrocytic gliosis. Morphometry three months after perinatal asphyxia showed significant reduction of corpus callosum in asphyctic brains. Patchy myelination deficits were found in hippocampal fimbriae and cerebellum, lobulus L 8, accompanied by reduced axonal density. Hypothalamus and striatum did not show any myelination deficit. Up to now only short term effects of perinatal asphyxia on myelination have been reported and this communication reveals long-term myelination deficit in three brain regions after three months following perinatal asphyxia. As myelination deficit was regularly accompanied by reduction of neurofilament immunoreactivity, we suggest that white matter lesions are paralleling grey matter damage, a subject still controversial in pathophysiology of brain damage in perinatal asphyxia. PMID- 11065183 TI - Pharmacological modification of the serotonergic transmitter system and beta-N acetylhexosaminidase activity in rats. AB - The enzyme activity and activation energy of plasma beta-N-acetylhexosaminidase (Hex) was determined in rats whose serotonergic system had been pharmacologically altered. In the group of animals treated with 5-hydroxytryptophan, in the different dissected brain regions (brain stem, cortex and hippocampus) significantly higher levels of serotonin and 5-hydroxyindolacetic acid were found, and significantly lower in the group treated with p-chlorophenylalanine, than in the control group. In the total number of animals studied (n = 21), a statistically significant correlation was found between the plasma concentration of 5-hydroxyindolacetic acid and the levels of this metabolite in the different brain regions (p < 0.001). No significant differences were found for the activity of Hex in the plasma, or for its activation energy, which is a marker of its isoenzyme composition, among the three groups of animals. The results obtained using our experimental model in rats do not confirm the hypothesis of other authors who suggest that the Hex responds secondary to increases or decreases of serotonin turnover, and could be a biological test to monitor the serotonin status in psychiatric patients. PMID- 11065184 TI - Homeostatic role of the active transport in elimination of [3H]benzylpenicillin out of the cerebrospinal fluid system. AB - Cerebral acidic metabolites and penicillin are organic anions which can be carried by active transport into capillaries of the central nervous system (CNS). However, it is generally believed that these metabolites are mainly delivered from CNS to cerebrospinal fluid (CSF) and eliminated by CSF circulation over cortex and its absorption into dural venous sinuses. To test this hypothesis we studied fate of penicillin ([3H]benzylpenicillin) in the CSF under control conditions and when its active transport was blocked by probenecid. After application of penicillin into cisterna magna of control dogs, it is distributed only in traces to lumbar, ventricular and cortical CSF. However, when active transport of penicillin across capillary wall is blocked by probenecid, its disappearance from cisterna is slowed down and its distribution is greatly enhanced so that at 300 min penicillin concentrations in cisternal, lumbar and cortical CSF approach or equal each other. Disappearance of penicillin from cisternal CSF shows a single exponential course (half-time 30 min) in control, while in probenecid pretreated dogs this is a slow multiexponential process. The results indicate that the active transport across capillary wall in CNS, but not generally postulated unidirectional CSF circulation over cortex and its absorption into dural venous sinuses, is instrumental in elimination of cerebral acidic metabolites and in such a way homeostasis in brain and cerebrospinal fluid is maintained. PMID- 11065185 TI - MCI-186: further histochemical and biochemical evidence of neuroprotection. AB - The bioactivity of 3-methyl-1-phenyl-pyrazolin-5-one (MCI-186) was examined based on histochemical changes in drastic global ischemic rat brains. Rats with mean arterial blood pressure reduction were subjected to 60 min cerebral ischemia/80 min reperfusion. Infusion of MCI-186 at 3.0 mg/Kg reduced brain infarction from 21 +/- 4% (saline control, n= 15) to 11 +/- 3% (n=16, p<0.05). By comparison, infusion of up to 20 mg/Kg propyl galalate (PG)--a well documented antioxidant- produced an infarct percentage of 14 +/- 5% (n=8), close to the saline control. Biochemically, the neuroprotective effect of MCI-186 was demonstrated by diminishing the release of creatine kinase (CK) in serum from 3363 +/- 608 U/L (n=14) in saline control to 1989 +/- 293 U/L (n= 15) in MCI group (p<0.05), while PG did not lower the activity of CK significantly. MCI-186 behaves as a free radical scavenger by suppressing the formation of superoxide anion in xanthine oxidase (XO)-hypoxanthine (HP) system (p<0.05). Our data supported our contention that MCI-186 has potent anti-stroke effect with antioxidant activities. PMID- 11065186 TI - Effects of econazole on receptor-operated and depolarization-induced contractions in rat isolated aorta. AB - In our previous study, econazole caused a decrease in serum nitrite levels in septic mice in vivo, but it enhanced the mortality rate. The aim of the study was to investigate the in vitro effects of econazole on receptor-operated and depolarization-induced contractions on endothelium-intact and -denuded rat isolated aorta. Econazole (0.1, 1 and 10 microM) significantly inhibited receptor operated (phenylephrine, Phe) and depolarization (KCl)-induced contractions of endothelium-intact or -denuded rings in a noncompetitive and concentration dependent manner. Removal of endothelium changed the pD'2 values only for KCl induced responses. The pD'2 values of L-type calcium channel blocker nifedipine were significantly higher than the econazole on Phe concentration-response curves in endothelium-intact and -denuded rings. Econazole caused a biphasic response in precontracted by Phe or KCl in endothelium-intact and -denuded rings, first a transient contraction following sustained relaxation. Removal of endothelium did not affect the contractile responses induced by Phe. The contractile responses induced by 10 microM econazole in the KCl-precontracted rings were antagonized by the treatment of alpha-adrenergic receptor antagonist, phentolamine (10 microM). Deendothelization was significantly increased the IC50 values of econazole obtained from Phe- and KCl-precontractions. The relaxations induced by 10 microM econazole in endothelium-intact rings precontracted with Phe or KCl were not changed by NO synthase inhibitor, L-N(G)-nitroarginine (100 microM). The IC50 values of econazole were significantly higher than nifedipine in endothelium intact and -denuded rings. These results suggest that econazole is a noncompetitive antagonist on alpha1-adrenoceptor-mediated and depolarization induced contractions in rat isolated aorta by inhibiting Ca2+ entry through L type calcium channels, and the endothelium seems to modulate vascular responses induced by this agent. The vascular effects of econazole may limit the usage of this agent in septic shock. PMID- 11065187 TI - Cu(II) and Zn(II) complexes with hyaluronic acid and its sulphated derivative. Effect on the motility of vascular endothelial cells. AB - With the aim of improving the compatibility of biomaterials to be used for the construction of cardiovascular prosthesis, we have designed bioactive macromolecules resulting from chemical modifications of hyaluronic acid (Hyal). The stability constants of Cu(II) and Zn(II) complexes with the sulphated derivative of hyaluronic acid (HyalS3.5) were evaluated. Two different complexes have been found for each metal ion, CuL, Cu(OH)2L and ZnL, Zn(OH)2L (L means the disaccharide unit of the ligands) in aqueous solution at 37 degrees C. The dihydroxo Cu(II) complex was present in high percentage at pH=7.4. On the contrary, the Zn(II) ion was present with a relatively low percentage of both complexes. The ability to stimulate endothelial cell adhesion and migration was evaluated for Hyal, HyalS3.5 and their complexes with Cu(II) and Zn(II) ions. The results revealed that Hyal and [Cu(OH)2HyalS3.5](4.5)- induced cell adhesion, while [ZnHyalS3.5](2.5)- and [Zn(OH)2HyalS3.5](4.5)- inhibited the process. The chemotactic activity of increasing concentrations of the above complexes was also evaluated, demonstrating that [Cu(OH)2HyalS3.5](4.5)- complex at 1 microM concentration was the most active in inducing cell migration. These results have been also strengthened by analysing adherent cell migration in agarose. In conclusion, sulphated hyaluronic acid coordinated to Cu(II) seems to be a promising matrix molecule for the construction of cardiovascular prosthesis. PMID- 11065188 TI - Mathematical modelling of kinetics of adenosine 5'-triphosphate hydrolysis catalyzed by Zn2+ ion in the pH range 7.1-7.4. AB - Kinetic data on hydrolysis of ZnATP2- complexes confirm the enzyme-like mechanism of the reaction. The whole sequence of steps for the formation and transformations of the intermediates is established by numerical modelling in a wide range of concentrations (4x10(-4)-0.3 M) in the pH range 7.1-7.4. The rates of active center formation and appropriate equilibria are governed by H+ transfer from coordinated water with formation of hydrogen bond between the (N1) atom of the second ZnATP2- molecule and the gamma-phosphate moiety of the first ZnATP2- molecule. The rate and equilibrium constants are higher in trimeric associates as compared to dimeric ones. Among the steps of ADP formation in the pH-independent channel, H+ transfer from the hydrogen bond with O(-)-Pgamma of ZnATP2- to the hydrogen bond with O(-)-Pbeta of ZnADP- forming in the course of general base catalysis is the rate determining step. It is followed by the rapid and reversible substitution of ligand H2PO4- by H2O in the Zn2+ coordination sphere. Hydrogen bond participation leads to reversible ADP formation. AMP is shown to be formed also via associates, and the conformation transformation determines the induction period. The induction period decreases as the concentration of ZnATP2- increases. The rate and equilibrium constants of all steps are evaluated and variation of the intermediate concentrations in the course of hydrolysis is presented. PMID- 11065189 TI - Ionic strength and pH effect on the Fe(III)-imidazolate bond in the heme pocket of horseradish peroxidase: an EPR and UV-visible combined approach. AB - The effects of chloride, dihydrogenphosphate and ionic strength on the spectroscopic properties of horseradish peroxidase in aqueous solution at pH=3.0 were investigated. A red-shift (lambda=408 nm) of the Soret band was observed in the presence of 40 mM chloride; 500 mM dihydrogenphosphate or chloride brought about a blue shift of the same band (lambda=370 nm). The EPR spectrum of the native enzyme at pH 3.0 was characterized by the presence of two additional absorption bands in the region around g=6, with respect to pH 6.5. Chloride addition resulted in the loss of these features and in a lower rhombicity of the signal. A unique EPR band at g=6.0 was obtained as a result of the interaction between HRP and dihydrogenphosphate, both in the absence and presence of 40 mM Cl . We suggest that a synergistic effect of low pH, Cl- and ionic strength is responsible for dramatic modifications of the enzyme conformation consistent with the Fe(II)-His170 bond cleavage. Dihydrogenphosphate as well as high chloride concentrations are shown to display an unspecific effect, related to ionic strength. A mechanistic explanation for the acid transition of HRP, previously observed by Smulevich et al. [Biochemistry 36 (1997) 640] and interpreted as a pure pH effect, is proposed. PMID- 11065190 TI - HgCl2 disrupts the structure of the human erythrocyte membrane and model phospholipid bilayers. AB - The structural effects of Hg(II) ions on the erythrocyte membrane were studied through the interactions of HgCl2 with human erythrocytes and their isolated resealed membranes. Studies were carried out by scanning electron microscopy and fluorescence spectroscopy, respectively. Hg(II) induced shape changes in erythrocytes, which took the form of echinocytes and stomatocytes. This finding means that Hg(II) locates in both the outer and inner monolayers of the erythrocyte membrane. Fluorescence spectroscopy results indicate strong interactions of Hg(II) ions with phospholipid amino groups, which also affected the packing of the lipid acyl chains at the deep hydrophobic core of the membrane. HgCl2 also interacted with bilayers of dimyristoylphosphatidylcholine and dimyristoylphosphatidylethanolamine, representative of phospholipid classes located in the outer and inner monolayers of the erythrocyte membrane, respectively. X-ray diffraction indicated that Hg(II) ions induced molecular disorder to both phospholipid bilayers, while fluorescence spectroscopy of dimyristoylphosphatidylcholine large unilamellar vesicles confirmed the interaction of Hg(II) ions with the lipid polar head groups. All these findings point to the important role of the phospholipid bilayers in the interaction of Hg(II) on cell membranes. PMID- 11065191 TI - Interactions of metal ions with a 2,4-diaminopyrimidine derivative (trimethoprim). Antibacterial studies. AB - The interaction of copper (II), zinc(II) and cadmium(II) with Trimethoprim (2,4 diamino-5-(3',4',5'-trimethoxybenzyl) pyrimidine) has been studied. The crystal structures of [Zn(Trim)2Cl2] (2) and [Cd(Trim)Cl2(CH3OH)]n (4) are reported. Compound (2) exhibits a distorted tetrahedral environment around the metal center and crystallizes in the triclinic space group P1 with a=10.2397(6), b=10.4500(6), c=16.3336(16) A, alpha=96.141(8), beta=106.085(5), gamma=96.551(5) degrees and Z=2. In complex (4), the Cd(II) centers are bridged sequentially by two chlorine ions to form infinite chains and present a six-coordinated environment; the compound crystallizes in the monoclinic P2(1)/C space group with a=13.958(5), b=7.532(2), c=18.390(2) A, alpha=90, beta=97.32(5), gamma=90 degrees and Z=4. In both structures the Trimethoprim acts as a monodentate ligand through the pyrimidinic nitrogen N(1) atom. The characterization of the Cu(Trim)2(CH3O)(ClO4) complex through EPR and magnetic measurements suggests a binuclear or polinuclear nature, with bridging methoxo groups. The complexes were screened for their activity against several bacteria, showing activity similar to that of trimethoprim. PMID- 11065192 TI - Redox and fungicidal properties of phthalocyanine metal complexes as related to active oxygen. AB - Some chemical and fungicidal effects of 20 phthalocyanines of Co, Fe, Cu, and Al were studied. Under dark conditions, these complexes reduced nitroblue tetrazolium in the presence of KCN, accelerated the autoxidation of ascorbate or hydroquinone and decomposed hydrogen peroxide. In the later reaction, hydroxyl radical was generated as evidenced with the deoxyribose assay. The inhibition by superoxide dismutase and catalase of catalyzed autoxidation of ascorbate suggests the participation of superoxide anion-radical and hydrogen peroxide in the reaction. Most complexes were toxic to the fungus Magnaporthe grisea which causes blast disease of rice. The toxicity was enhanced by light being diminished by antioxidant reagents sequestering active oxygen species. Some complexes (including nontoxic ones), after 1-day contact with a leaf surface of the disease susceptible rice cultivar, induced the fungitoxicity of leaf diffusate. This toxicity was also light-activated and sensitive to antioxidant reagents. Several complexes, when added to inocula, decreased 2-3 times the frequency of the compatible symptoms of the blast. It is suggested that in planta, the dark redox activity of phthalocyanines along with their photosensitization promote the generation of active oxygen, which damages the parasite and, therefore, favors disease resistance. PMID- 11065193 TI - Oxymyohemerythrin: discriminating between O2 release and autoxidation. AB - Myohemerythrin (Mhr) is a non-heme iron O2 carrier (with two irons in the active site) that is typically found in the retractor muscle of marine 'peanut' worms. OxyMhr may either release O2, or undergo an autoxidation reaction in which hydrogen peroxide is released and diferric metMhr is produced. The autoxidation reaction can also be promoted by the addition of certain anions to Mhr solutions. This work, using recombinant Themiste zostericola Mhrs, contrasts the results of environmental effects on these reactions. For the O2 release reaction, deltaVdouble dagger(21.5 degrees C) = +28+/-3 cm3 mol(-1), deltaHdouble dagger(1 atm) = +22+/-1 kcal mol(-1), and deltaSdouble dagger(1 atm) = +28+/-4 eu. The autoxidation reaction (pH 8.0, 21.5 degrees C, 1 atm) displays different kinetic parameters: deltaVdouble dagger = -8+/-2 cm3 mol(-1), deltaHdouble dagger = +24.1+/-0.7 kcal mol(-1), and deltaSdouble dagger = +1+/-1 eu. Autoxidation in the presence of sodium azide is orders of magnitude faster than solvolytic autoxidation. The deltaVdouble dagger parameters for azide anation and azide assisted autoxidation reaction are +15+/-2 and +59+/-2 cm3 mol(-1), respectively, indicating that the rate-limiting steps for the Mhr autoxidation and anation reactions (including O2 uptake) are not associated with ligand binding to the Fe2 center. The L103V and L103N oxyMhr mutants autoxidize approximately 10(3)-10(5) times faster than the wild-type protein, emphasizing the importance of leucine 103, which may function as a protein 'gate' in stabilizing bound dioxygen. PMID- 11065194 TI - Aluminum speciation studies in biological fluids. Part 6. Quantitative investigation of aluminum(III)-tartrate complex equilibria and their potential implications for aluminum metabolism and toxicity. AB - Recent epidemiological studies have confirmed the existence of a correlation between aluminum level in low-silica drinking water and prevalence of Alzheimer's disease. Also, oral aluminum-based phosphate binders and antacids may induce acute aluminum toxicity. Whatever the source of the metal ingested, its bioavailability is a function of the chemical forms under which it occurs in the gastrointestinal tract, i.e. of the ligands with which the Al3+ ion may associate. Dietary acids in particular can favor the bioavailability of aluminum in different ways: by increasing its solubility, by complexing it into neutral species, and/or by acting indirectly on its absorption process. Among these, tartaric acid is commonly found in fruits and in industrial foods and drinks, and may therefore be ingested together with environmental or/and therapeutic aluminum. The present work examines its potential influence on aluminum bioavailability. Firstly, Al(III)-tartrate complex formation constants have been determined under physiological conditions (37 degrees C, 0.15 M NaCl). Then these constants have been used to simulate the influence of tartrate on aluminum speciation in different gastrointestinal situations in which phosphate was also taken into account. Under normal conditions of aluminum contamination, tartrate is expected to keep the metal soluble throughout the whole pH range of the small intestine, which is likely to enhance its bioavailability. Even at low concentrations, tartrate also gives rise to two neutral complexes that span over the 1.5-7.5 pH interval, a phenomenon that is aggravated by increased aluminum levels as may result from aluminum hydroxide therapy. The co-occurrence of dietary phosphate reduces the fraction of aluminum neutralized by tartrate under normal conditions, but this effect quickly decreases with increasing aluminum doses. Even the therapeutic use of aluminum phosphate is not expected to be totally safe in the presence of tartaric acid. As plasma simulations show that no aluminum mobilization can be expected from tartrate that could enhance aluminum excretion, avoiding ingestion of tartaric acid during any form of aluminum-based therapy appears advisable. PMID- 11065195 TI - Effects of monofunctional platinum binding on the thermal stability and conformation of a self-complementary 22-mer. AB - We investigated the effect of various monofunctional platinum complexes on the thermal stability and conformation of a self-complementary 22-mer duplex oligonucleotide by means of CD and UV melting profiles. We studied several families of triamine complexes of the general formula PtA2AmCl where A2=(NH3)2 and ethylenediamine and where Am=N1-4-methyl-pyridine, N7-guanosine, and 9-ethyl guanine. Platination by the N1-4-methyl-pyridine and 9-ethyl-guanine complexes led to a decrease in the Tm of the oligonucleotide by 2-11.5 degrees C while platination with the N7-guanosine complexes led to a rise in the melting temperature of the oligonucleotides by 4.5 degrees C. A similar inverse correlation between the two groups of platinum compounds was found in the CD spectra. In all cases, the cis isomer had a more pronounced effect on both the melting curve and the CD spectrum. The cis isomer was found to have a more destabilizing effect than its trans counterpart. This indicates that the cis geometry in fact forces a greater structural constraint on the backbone of the double helix. We have also found that the sugar of the guanosine has a significant influence on both the Tm and CD spectra; the sugar moiety contributes to the stability of the double helix, probably through the formation of hydrogen bonds. PMID- 11065196 TI - Endogenous morphine--another component and biological modifier of the response to surgical injury? PMID- 11065197 TI - Pharmacokinetic studies of neuromuscular blocking agents: good clinical research practice (GCRP). AB - In September 1997, an international consensus conference on standardization of studies of neuromuscular blocking agents was held in Copenhagen, Denmark. Based on the conference, a set of guidelines for good clinical research practice (GCRP) in pharmacokinetic studies of neuromuscular blocking agents is presented. Guidelines include: design of the study; relevant patient groups to investigate; test drug administration, sampling and analysis; pharmacokinetic analysis; pharmacokinetic/pharmacodynamic modeling; population pharmacokinetics; statistics; and presentation of pharmacokinetic data. The guidelines are intended to aid those working in this research area; it is hoped that they will assist researchers, editors of scientific papers, and pharmaceutical companies in improving the quality of pharmacokinetic studies. PMID- 11065198 TI - Non-opioid postoperative analgesia. AB - Although significant improvement has been made in the treatment of pain in the postoperative period, many patients still experience unnecessary discomfort resulting in distress, higher morbidity and prolonged stay in hospital. The standard pillar of postoperative treatment of severe pain is the use of opioids. However, adverse reactions to opioids make their use unfavourable. A better understanding of the pathophysiology of pain has helped clinicians to a more balanced approach to postoperative pain treatment. The development of the multimodal approach to postoperative analgesia, with the use of different drugs acting via different routes to give good analgesia, with minimal side-effects, represents a major development in the treatment of postoperative pain. Early, aggressive mobilisation and feeding must follow in order to restore normal conditions quickly. Alternatives to opioids should be used as extensively as possible. Local anaesthesia, used as regional blocks or as wound infiltration, is most beneficial. Paracetamol has good basic analgesic properties, and should probably be used in dosages higher than recommended today. The combination with a NSAID results in better and longer-lasting analgesia. The intravenous form propacetamol will increase the possibilities of its use. The new concept of selective COX-2 inhibiting NSAIDs will result in analgesic and anti-inflammatory drugs with fewer side-effects. The well-known inexpensive group of corticosteroids have good analgesic and anti-emetic properties, and are especially interesting to use in patients who do not tolerate NSAIDs. The alpha2 receptor agonists like clonidine, when administered epidurally or intrathecally, are useful adjuncts, but their adverse effects on sedation and hypotension limit their use. NMDA-receptor antagonists are of limited value in the postoperative period. Adenosine and neostigimine are still on a research level but may lead to new, clinically useful analgesic drugs. In the future, cannabinoids, cholecystokinin-receptor antagonists and neurokinin-1 antagonists may become important analgesic drugs. PMID- 11065199 TI - Endogenous morphine is produced in response to cardiopulmonary bypass in neonatal pigs. AB - BACKGROUND: Cardiopulmonary bypass (CPB) is associated with a systemic inflammatory response. Endogenous morphine production has previously been demonstrated in humans after cardiac surgery with CPB. It has been hypothesized that morphine plays a role as an anti-inflammatory mediator in the systemic inflammatory response. The aim of this study was to investigate if the CPB procedure in itself elicits an endogenous morphine production in neonatal pigs. METHODS: Endogenous morphine production was measured in arterial blood in piglets exposed to sternotomy alone (sham group, n=10) or sternotomy and CPB (n=10). Blood samples were obtained immediately after the induction of anaesthesia, at the end of CPB and 4 h later. Morphine in arterial blood was detected by radioimmunoassay and confirmed by gas chromatography mass spectrometry. RESULTS: Animals undergoing CPB showed detectable endogenous morphine concentrations immediately after CPB, with increased concentrations postoperatively. There was no measurable morphine production in the sham operated pigs. CONCLUSION: The CPB procedures elicits an endogenous morphine production in neonatal pigs. This morphine response is analogous to the previously demonstrated response in patients subjected to cardiac surgery and CPB. PMID- 11065200 TI - Improved cerebral blood supply and oxygenation by aortic balloon occlusion combined with intra-aortic vasopressin administration during experimental cardiopulmonary resuscitation. AB - BACKGROUND: Intravenous administration of vasopressin during cardiopulmonary resuscitation (CPR) has been shown to improve myocardial and cerebral blood flow. Aortic balloon occlusion during CPR may also augment myocardial and cerebral blood flow and can be used as a central route for the administration of resuscitative drugs. We hypothesized that, as compared with intravenously administered vasopressin, the administration of this drug above the site of an aortic balloon occlusion would result in a greater increase in cerebral perfusion and oxygenation during CPR and after restoration of spontaneous circulation (ROSC). METHODS: Twenty piglets were subjected to 5 min of ventricular fibrillation followed by 8 min of closed-chest CPR and were treated with 0.4 U kg(-1) boluses of vasopressin intravenously (the IV-vasopressin group with sham aortic balloon) or above the site for an aortic balloon occlusion (the balloon vasopressin group). The aortic balloon catheter was inflated in the latter group 1 min after commencement of CPR and was deflated within 1 min after ROSC. Systemic blood pressures, cerebral cortical blood flow, cerebral tissue pH and PCO2 were monitored continuously and the cerebral oxygen extraction ratio was calculated. RESULTS: During CPR, arterial blood pressure and cerebral perfusion pressure were greater in the balloon-vasopressin group, as compared with the IV vasopressin group. These pressures did not differ between the groups after ROSC. Cerebral cortical blood flow was not significantly greater in the balloon vasopressin group during CPR, whereas significantly higher cortical blood flow levels were recorded after ROSC. Cerebral tissue pH decreased in the IV vasopressin group during the post-resuscitation hypoperfusion period. In contrast, decreasing pressures during the hypoperfusion period did not result in increasing tissue acidosis in the balloon-vasopressin group. CONCLUSIONS: During CPR, intra-aortic vasopressin combined with aortic balloon occlusion resulted in significantly greater perfusion pressures but not in greater cerebral cortical blood flow. After ROSC, however, a greater increase in cortical blood flow was recorded in the balloon-vasopressin group, even though the aortic balloon was deflated and perfusion pressures did not differ between the groups. This suggests that vasopressin predominantly gives vasoconstrictive effects on cerebral cortical vessels during CPR, but results in cerebral cortical vasodilatation after ROSC. PMID- 11065201 TI - Dynamic evaluation of fluid shifts during normothermic and hypothermic cardiopulmonary bypass in piglets. AB - BACKGROUND: Edema, generalized overhydration and organ dysfunction commonly occur in patients undergoing open-heart surgery using cardiopulmonary bypass (CPB) and induced hypothermia. Activation of inflammatory reactions induced by contact between blood and foreign surfaces are commonly held responsible for the disturbances of fluid balance ("capillary leak syndrome"). We used an online technique to determine fluid shifts between the intravascular and the interstitial space during normothermic and hypothermic CPB. METHODS: Piglets were placed on CPB (fixed pump flow) via thoracotomy in general anesthesia. In the normothermic group (n=7), the core temperature was kept at 38 degrees C prior to and during 2 h on CPB, whereas in the hypothermic group (n=7) temperature was lowered to 28 degrees C during bypass. The CPB circuit was primed with acetated Ringer's solution. The blood level in the CPB circuit reservoir was held constant during bypass. Ringer's solution was added when fluid substitution was needed (falling blood level in the reservoir). In addition to invasive hemodynamic monitoring, fluid input and losses were accurately recorded. Inflammatory mediators or markers were not measured in this study. RESULTS: Cardiac output, s electrolytes and arterial blood gases were similar in the two groups in the pre bypass period. At start of CPB the blood level in the machine reservoir fell markedly in both groups, necessitating fluid supplementation and leading to a markedly reduced hematocrit. This extra fluid need was transient in the normothermic group, but persisted in the hypothermic animals. After 2 h of CPB the hypothermic animals had received 7 times more fluid as compared to the normothermic pigs. CONCLUSION: We found strong indications for a greater fluid extravasation during hypothermic CPB compared with normothermic CPB. The experimental model using the CPB-circuit reservoir as a fluid gauge gives us the opportunity to study further fluid volume shifts, its causes and potential ways to optimize fluid therapy protocols. PMID- 11065202 TI - Surgical stress induces acute coronary release of tissue-type plasminogen activator in the pig. AB - BACKGROUND: Tissue-type plasminogen activator (t-PA) is an endothelium derived key enzyme in the initiation of endogenous fibrinolysis. Acute regulated release of active t-PA occurs within minutes in response to threatening thrombotic vessel occlusion. The aim of this study was to investigate the impact of surgical stimulation on the kinetics of t-PA release in the coronary vascular bed in the pig. METHODS: In anaesthetised pigs (n=16), arterio-venous concentration gradients of t-PA, and plasma flows (retrograde thermodilution) were obtained across the coronary vascular bed before (control) and at 1, 3, 5 and 10 min after sternotomy. RESULTS: At control, no significant coronary net flux (release or uptake) of t-PA was observed, while sternotomy induced a rapid net release of total t-PA (132.6 ng x min(-1)), with an associated increase in active t-PA (93.6 ng x min(-1)). This response, evident already after 1 min, showed a peak at 5 min and returned towards baseline levels within 10 min. No concurrent alterations in aortic levels of active t-PA were found and haemodynamic variables were unaltered. CONCLUSION: The rapidly increasing and transient net coronary release of t-PA after sternotomy suggests that the endothelium actively promotes local endogenous fibrinolysis during surgery. Such events could reflect a dynamic responsiveness to protect the coronary circulation during stress. PMID- 11065203 TI - Effects of remifentanil on neutrophil adhesion, transmigration, and intercellular adhesion molecule expression. AB - BACKGROUND: Anaesthetic drugs are used for pain therapy and anaesthesia. Neutrophils play a significant role during the process of inflammation. The aim of the current study was to investigate the effects of remifentanil and fentanyl on neutrophil migration through endothelial cell monolayers, and on adhesion molecule expression. METHODS: After isolation of polymorphonuclear neutrophils (PMNL) we used a currently described migration assay. PMNL and/or endothelial cell monolayers (ECM) were pre-treated with remifentanil using clinically relevant, as well as higher and lower concentrations or relevant concentrations of fentanyl. RESULTS: Concentrations of remifentanil (50 ng/mL) similar to the relevant plasma concentration were able to inhibit PMNL migration through ECM significantly (migration compared to the control 82+/-7% SD; P<0.05), when both cell types were treated with the synthetic narcotic remifentanil. Fentanyl (30 ng/mL) showed a stronger inhibitory effect (migration compared to the control 67+/-9.2%; P<0.05). Endothelial cell adhesion molecule expression was reduced after either remifentanil or fentanyl. CONCLUSION: The results of the present investigation indicate that remifentanil influences interaction of ECM against human neutrophils. Compared to fentanyl, remifentanil seems to exhibit minor inhibitory effects on neutrophil migration. PMID- 11065204 TI - Angiotensin II mesenteric and renal vasoregulation: dissimilar modulatory effects with nitroprusside. AB - BACKGROUND: The role of systemic arterial pressure for the vascular effects of angiotensin II (Ang II) and the interactions between Ang II and perfusion pressure-dependent local vascular control mechanisms are not well understood. This study addresses these aspects of exogenous Ang II in the mesenteric and renal regional circulations. METHODS: Ang II was infused in incremental doses (0 200 microg/h) in anesthetized instrumented pigs (n=10). Renal and portal blood flows were measured by perivascular ultrasound. In the second part of the study, sodium nitroprusside (SNP) was infused at doses titrated to keep mean arterial pressure constant, in spite of concurrent Ang II administration. RESULTS: Powerful dose-dependent vasoconstrictions by Ang II were found in renal and mesenteric vascular beds (at highest Ang II doses vascular resistances increased by 109% and 88% respectively). Ang II-induced vasoconstriction was fully inhibited in the mesenteric, but not in the renal circulation, during conditions of constant mean arterial pressures achieved by SNP infusion. CONCLUSIONS: Mesenteric, but not renal, vasoconstriction by Ang II was inhibited by pharmacological maintenance of perfusion pressure. This could reflect differences between these vascular beds as regards the importance of co-acting myogenic pressure-dependent vasoconstriction. Alternatively, as the drug chosen for pressure control, sodium nitroprusside, serves as a nitric oxide donor, the relative balance between nitric oxide-mediated vasodilation and Ang II-induced vasoconstriction could have regional differences. PMID- 11065205 TI - Cognitive dysfunction 1-2 years after non-cardiac surgery in the elderly. ISPOCD group. International Study of Post-Operative Cognitive Dysfunction. AB - BACKGROUND: Postoperative cognitive dysfunction (POCD) is a well-recognised complication of cardiac surgery, but evidence of POCD after general surgery has been lacking. We recently showed that POCD was present in 9.9% of elderly patients 3 months after major non-cardiac surgery. The aim of the present study was to investigate whether POCD persists for 1-2 years after operation. METHODS: A total of 336 elderly patients (median age 69 years, range 60-86) was studied after major surgery under general anesthesia. Psychometric testing was performed before surgery and at a median of 7, 98 and 532 days postoperatively using a neuropsychological test battery with 7 subtests. A control group of 47 non hospitalised volunteers of similar age were tested with the test battery at the same intervals. RESULTS: 1-2 years after surgery, 35 out of 336 patients (10.4%, CI: 7.2-13.7%) had cognitive dysfunction. Three patients had POCD at all three postoperative test sessions (0.9%). From our definition of POCD, there is only a 1:64000 likelihood that a single subject would have POCD at all three test points by chance. Logistic regression analysis identified age, early POCD, and infection within the first three postoperative months as significant risk factors for long term cognitive dysfunction. Five of 47 normal controls fulfilled the criteria for cognitive dysfunction 1-2 years after initial testing (10.6%, CI: 1.8-19.4%), i.e. a similar incidence of age-related cognitive impairment as among patients. CONCLUSION: POCD is a reversible condition in the majority of cases but may persist in approximately 1% of patients. PMID- 11065206 TI - Droperidol and 5-HT3-receptor antagonists, alone or in combination, for prophylaxis of postoperative nausea and vomiting. A meta-analysis of randomised controlled trials. AB - BACKGROUND: Droperidol and 5-HT3-receptor antagonists are among the most potent antiemetics to prevent postoperative nausea and vomiting (PONV). Combinations of these drugs have been used to increase the efficacy of antiemetic treatment. However, so far the quantitative effect of this combination has not been evaluated systematically. METHODS: Results from randomised controlled trials investigating the efficacy of 5-HT3-receptor antagonists or droperidol alone versus the combination of both drugs to prevent PONV were included in a meta analysis. Studies were systematically searched using Medline, EMBASE, the Cochrane-Library, and by manually screening the reference lists of matching review articles and current issues of locally available peer-reviewed anaesthesia journals. Seven papers with data on granisetron published by Fujii and co-workers were not considered. The main end point in each study was defined as occurrence of nausea, retching, or vomiting within 6 h ("early PONV") and within 48 h ("late PONV") after surgery. The relative risks (RR) and the numbers needed to treat (NNT) of the pooled data with their corresponding 95% confidence intervals (given in parentheses) were calculated using a random effects model. RESULTS: Eight studies with 881 patients (adults: n=801; children (mean age: 8 yr): n=80) were included in the analysis. Droperidol was applied to 340 patients, 5-HT3-receptor antagonists to 198, and 343 were treated with a combination of both drugs. Seven out of these eight studies reported increased antiemetic efficacy of the combination group compared with the single drugs (droperidol and 5-HT3-receptor antagonists respectively). However, in none of the trials did this difference reach statistical significance. When a meta-analytic analysis based on these results was performed the combination of droperidol with a 5-HT3-receptor antagonist was not associated with a significantly increased antiemetic efficacy. In 12 to 13 patients a 5-HT3-receptor antagonist has to be added to droperidol prophylaxis to prevent one additional patient from PONV who would have had suffered from PONV when treated with droperidol alone (RR "early PONV": 1.52 (0.95-2.44); RR "late PONV": 1.24 (0.89-1.74)). Similar results were obtained when the antiemetic effect of adding droperidol to a prophylaxis with 5-HT3 receptor antagonists was analysed. In this case 10 to 12 patients have to be treated with the 5-HT3-droperidol combination instead of with a 5-HT3-receptor antagonist alone to prevent one additional patient from PONV (RR "early PONV": 1.55 (0.68-3.52); RR "late PONV": 1.29 (0.77-2.17)). There were no reports of an increased incidence of adverse effects. CONCLUSION: The data on the combination of droperidol with 5-HT3-receptor antagonists suggest that there is a trend towards increased efficacy of the combination therapy compared to the single drugs. However, so far there are insufficient data to recommend this combination treatment for prophylaxis. PMID- 11065207 TI - Does efficacy of seal and fibreoptic view change during anaesthesia with the laryngeal mask airway: a comparison of oxygen and oxygen-nitrous oxide gas mixtures. AB - BACKGROUND: We test the hypothesis that oropharyngeal leak pressure (OLP) and fibreoptic position (FP) are stable for the laryngeal mask airway (LMA) during anaesthesia with and without nitrous oxide. METHODS: Forty paralysed anaesthetised patients (ASA 1-2, aged 18-80) were randomly allocated to receive 100% oxygen (O2 group) or 33% oxygen in nitrous oxide (O2-N2O group) for maintenance. In vivo intracuff pressure (CP) using a size 5 LMA was adjusted to 60 cm H2O. OLP, FP and CP were measured every 5 min for 30 min. RESULTS: CP was higher in the O2-N2O group than the O2 group, other than at time zero (all: P<0.0001). CP increased every 5 min in the O2-N2O group (all: P<0.0001), but not in the O2 group. There were no differences in OLP and FP between groups at any time. There were no significant changes in OLP or FP with time within each group. There were no changes in OLP greater than 1.5 cm H2O and no change in FP for any patient. CONCLUSION: We conclude that OLP and FP are stable for the LMA during anaesthesia lasting 30 min with or without nitrous oxide. PMID- 11065208 TI - No genotoxic effect of propofol in chinese hamster ovary cells: analysis by sister chromatid exchanges. AB - BACKGROUND: In spite of its high placental transfer, propofol is frequently used in general anesthesia and sedation during obstetric and gynecological surgery such as in vitro fertilization. This study investigated whether or not propofol has a genotoxic potential by the sister chromatid exchange assay in vitro. METHODS: Sister chromatid exchanges induced after exposure to propofol were measured in Chinese hamster ovary cells with and without metabolic activation. After propofol (0.2-20 microg ml(-1)) diluted dimethyl sulfoxide was applied for 2 h with or without S9 mix, the cells having been incubated for two metaphases (34 h) in the presence of 5'-bromo-2-deoxyuridine. N-nitrosodimethylamine and mitomycin C were used as positive controls with and without metabolic activation. The chromosomes were stained with the fluorescence plus Giemsa method, and then sister chromatid exchanges in 50 cells were counted for each concentration. RESULTS: Although increasing concentrations of propofol inhibited cell proliferation, no concentrations of propofol used in this study increased the sister chromatid exchange values, with and without metabolic activation. CONCLUSION: It was concluded that there was no indication, from the sister chromatid exchange assay in mammalian cells, of a genotoxic effect of propofol and its metabolites. PMID- 11065209 TI - Are electrocardiogram electrodes acceptable for electroencephalogram bispectral index monitoring? AB - BACKGROUND: The monitoring of electroencephalogram bispectral index (EEG-BIS) during anaesthesia reduces anaesthetic use and improves recovery. However, it also increases the direct costs of anaesthesia due to the need for special EEG electrodes. In the present study we tested the feasibility of less expensive electrocardiogram (ECG) electrodes for EEG-BIS monitoring. METHODS: In the first part of the study we compared skin-electrode impedances when EEG electrodes were used after alcohol swab pretreatment of skin to impedances when ECG electrodes were used after alcohol swab pretreatment with or without skin abrasion paste. In the second part of the study we evaluated the difference in parallel BIS values collected with two BIS monitors, using either ECG electrodes or EEG electrodes. In the third part of the study we compared parallel BIS values collected with two sets of EEG electrodes. RESULTS: Skin pretreatment with abrasion paste led to lower impedances with ECG electrodes than did alcohol swab pretreatment of skin with EEG electrodes. When the skin was pretreated with alcohol swab, higher impedances were measured with ECG electrodes than with EEG electrodes. In most patients, BIS values collected with ECG electrodes were also higher than those collected with adhesive EEG electrodes. The difference between parallel BIS values collected with two sets of adhesive EEG electrodes was smaller than the difference between BIS values collected with ECG and EEG electrode sets. CONCLUSION: Low skin-electrode impedances indicating reliable skin-electrode contact can be ensured with inexpensive pregelled ECG electrodes only if the skin is carefully prepared with both abrasion paste and alcohol. When only alcohol pretreatment of skin is used, the BIS values collected with EEG electrodes and ECG electrodes are not equal. EEG-BIS monitoring with pregelled ECG electrodes is recommended only if skin is prepared with abrasion paste before attaching the electrodes. PMID- 11065210 TI - Stroke following appendectomy under general anesthesia in a patient with basilar impression. AB - We report a boy who developed a vertebral stroke immediately after an appendectomy. Basilar impression was diagnosed eight years after this event when skull roentgenograms revealed basilar impression with high standing tip of the odontoid. We speculate that muscle relaxation and cervical hyperextension during intubation in the presence of basilar impression resulted in vertebral artery dissection and stroke. We suggest that patients with vertebral stroke and no obvious risk factors should be evaluated for the presence of malformations of the craniovertebral junction to be able to take precautions against excessive neck movement during intubation. PMID- 11065211 TI - Complications and technical aspects of jet ventilation for endolaryngeal procedures. PMID- 11065212 TI - Increase in neurogenic nitric oxide metabolism by endothelin-1 in mesenteric arteries from hypertensive rats. AB - We investigated, in mesenteric arteries from hypertensive rats (SHRs), the possible changes in neurogenic nitric oxide (NO) release produced by endothelin-1 (ET-1), and the mechanisms involved in this process. The contractile response induced by electrical field stimulation (EFS; 200 mA, 0.3 ms, 1-16 Hz, for 30 s) in deendotheliumized mesenteric segments was abolished by tetrodotoxin and phentolamine. The NO synthase inhibitor N(G)-nitro-L-arginine (L-NAME, 10 microM) increased the contractions caused by EFS. ET-1 enhanced the contraction induced by EFS, which was unaltered by the subsequent addition of L-NAME. The ETA antagonist-receptor BQ-123 (1 microM) inhibited the effect of ET-1 on EFS response, whereas the ETB antagonist-receptor BQ-788 (3 microM) partially blocked it, and the subsequent addition of L-NAME restored the contractile response in both cases. SOD (25 unit/ml) decreased the response to EFS, and the subsequent addition of L-NAME increased this response. ET-1 did not modify the decrease in EFS response induced by SOD, and the addition of L-NAME increased the response. None of these drugs altered the response to exogenous noradrenaline (NA) or basal tone except SOD, which increased the basal tone, an effect blocked by phentolamine (1 microM). In arteries preincubated with [3H]NA, ET-1 did not modify the tritium efflux evoked by EFS, which was diminished by SOD. ET-1 did not alter basal tritium efflux, whereas SOD significantly increased the efflux. These results suggest that EFS of SHR mesenteric arteries releases neurogenic NO, the metabolism of which is increased in the presence of ET-1 by the generation of superoxide anions. PMID- 11065213 TI - Interaction between L-type Ca2+ channels and sarcoplasmic reticulum in the regulation of vascular tone in isolated rat small arteries. AB - A dysfunction of the sarcoplasmic reticulum (SR) causes an increase in the myogenic tone of rat skeletal muscle small arteries (A(SK)), but not that of mesenteric small arteries (A(MS)). We hypothesized that the difference depends on the activity of voltage-dependent Ca2+ channels in these vessels. To test this, we measured the membrane potential of these vessels and examined ryanodine induced constrictions by manipulating the activity of voltage-dependent Ca2+ channels. The isolated vessels were cannulated to control the transmural pressure. To assess the vascular tone, the inner diameter was measured with a video-digitizing system. The membrane potential of A(SK) was more depolarized between 20 to 100 mm Hg of transmural pressure. A(MS) was not constricted by the Ca2+ channel agonist Bay K 8644 (1 nM(-1) microM) alone, but substantially constricted in the presence of ryanodine (1 microM). Ryanodine also augmented the KCl (20 mM)-induced constriction. In A(SK), the Ca2+ channel blocker nisoldipine fully dilated the ryanodine-induced constriction: however, the ryanodine-induced constriction was less susceptible to nisoldipine than was the myogenic and phenylephrine-induced constriction caused mainly by increased Ca2+ influx. In conclusion, the contribution of the SR function to Ca2+ metabolism depends on the activity of dihydropyridine-sensitive Ca2+ channels. The dysfunction of SR by ryanodine may impair the Ca2+ extrusion rather than increase Ca2+ influx in rat small arteries. PMID- 11065214 TI - Inhibition of 12(S)-hydroxyeicosatetraenoic acid (12-HETE) production suppressed the intimal hyperplasia caused by poor-runoff conditions in the rabbit autologous vein grafts. AB - The efficacy of OPC-29030, a newly developed inhibitor of 12(S) hydroxyeicosatetraenoic acid (12-HETE) production, was evaluated on intimal hyperplasia of experimental autologous vein grafts in a distal poor-runoff model and a hyperlipidemic model in rabbits. First, rabbits were divided into two groups, the distal poor-runoff group (PR group) and the hyperlipidemic group (HL group). After 4 weeks preparing the PR model and the HL model, the femoral vein was implanted into the ipsilateral femoral artery. Then they were subdivided into two groups, depending on the diet provided; diet group with 0.1% OPC-29030 (OPC 29030 group) and normal diet group (control group). At 4 weeks, the grafts were harvested, and intimal hyperplasia of the graft was measured with an ocular cytometer. Intimal cell proliferation was determined by bromodeoxyuridine (BrdU) incorporation at 2 weeks after surgery. In addition, the effect of OPC-29030 on the proliferation or migration of rat aortic smooth muscle cells in culture was investigated. In the in vivo study in the PR group, the intimal hyperplasia and the plasma 12-HETE levels in the OPC-29030 group were significantly inhibited, compared with those of the control group. However, in the HL group, the intimal hyperplasia in both the OPC-29030 and control groups showed a remarkable degree of intimal hyperplasia. There was no significant difference between those two groups. Furthermore, there was no significant difference in the plasma 12-HETE levels in the HL group irrespective of the presence of OPC-29030. The BrdU labeling index at 2 weeks after grafting was significantly lower in the OPC-29030 group compared with that in the control group in the PR group. In the in vitro study, OPC-29030 did not inhibit smooth muscle cell proliferation; however, OPC 29030 inhibited the migration. These results demonstrate the efficacy of OPC 29030 in reducing the degree of intimal hyperplasia under PR conditions, but not under hyperlipidemic conditions. The mechanism of reducing the intimal hyperplasia may be that OPC-29030 inhibited 12-HETE production, which did not inhibit proliferation while inhibiting migration of the smooth muscle cell. These results suggested the possible involvement of 12-HETE with the intimal hyperplasia under PR conditions. PMID- 11065215 TI - Differential pattern of endothelin-1-induced inotropic effects in right atria and left ventricles of the human heart. AB - In human right atrium, endothelin A (ET(A)) receptors couple to both inositol phosphate formation and inhibition of adenylylcyclase, whereas in human left ventricle, ET(A) receptors couple only to inositol phosphate formation. To find out whether this might be of functional relevance, we studied, in right atria obtained from 32 patients undergoing coronary bypass grafting without apparent heart failure, and in right atria and left ventricles from eight patients with end-stage heart failure (NYHA IV) undergoing heart transplantation, the effects of endothelin-1 (ET-1) on basal force of contraction or on force of contraction increased by 1 microM forskolin. ET-1 (0.1 microM) exerted a positive inotropic effect in atrial and ventricular tissue; this could be antagonized by the ET(A) receptor antagonist BQ 123, but not by the ET(B)-receptor antagonist BQ 788. In atrial, but not in ventricular tissue, this positive inotropic effect was preceded by a transient negative inotropic effect. This negative inotropic effect was inhibited by BQ 123, but not by BQ 788. It was significantly prolonged in forskolin-prestimulated atria, and was significantly larger in atria from failing hearts. We conclude that, because ET-1 inhibits adenylylcyclase and causes negative inotropic effects in atria but not in ventricles, adenylylcyclase inhibition might be responsible for the transient negative inotropic effect of ET 1. PMID- 11065216 TI - Restoration of flow-dependent coronary dilation by ACE inhibition improves papaverine-induced maximal coronary blood flow in hypertensive patients: demonstration that large epicardial coronary arteries are more than conductance vessels. AB - Restoration of flow-dependent coronary artery dilation by angiotensin-converting enzyme inhibition (ACEI) has been demonstrated in patients with hypertension. The aim of the present study was to evaluate whether dilation of conductance coronary arteries may alter maximal coronary blood flow (CBFmax) and minimal coronary resistance (CRmin) in hypertensive patients with reversible impairment of flow dependent coronary artery dilation. Thirteen hypertensive patients with angiographically normal coronary arteries and no other risk factors were studied. Cross-sectional areas (CSAs) of proximal and distal left anterior descending (LAD) coronary arteries were determined by quantitative angiography. Coronary flow velocity was recorded in the distal LAD with an intracoronary Doppler catheter. Estimates of coronary blood flow and resistance were calculated at rest and during maximal increase in blood flow induced by papaverine injected in the midportion of the LAD, both before and after ACEI. Flow-dependent dilation of the proximal LAD, abolished before ACEI, was restored after (26.7 +/- 11.2%; p < 0.001). The increase in CSA of the distal LAD exposed to papaverine was significantly higher after ACEI than before (from 33.4 +/- 20.5% to 51.5 +/- 23.4%; p < 0.001). After restoration of proximal LAD flow-dependent dilation, CBFmax was increased by +21.0 +/- 10.3% (p < 0.001), and CRmin was reduced by 19.3 +/- 9.5% (p < 0.001). Thus, dilation of epicardial coronary arteries participates substantially in the coronary resistance in hypertensive patients. Restoration of flow-dependent coronary artery dilation by ACEI may improve the ability of coronary circulation to deliver its maximal myocardial blood flow in hypertensive patients. PMID- 11065217 TI - Increased renal vasoconstriction and gene expression of cyclooxygenase-1 in renovascular hypertension. AB - Vascular responses to arachidonic acid (AA) in the renal circulation are increased in hypertensive rats. We have suggested that these differences are related to changes in AA metabolism. In this study we investigated the mechanism involved in the increased AA-induced renal vasoconstriction. We evaluated vascular renal reactivity in the isolated perfused kidney, cyclooxygenase activity, protein content, and mRNA expression of kidneys from sham operated and aortic coarctation rats. Bolus injection of AA (1, 2, 4, and 8 microg) increased perfusion pressure in a dose-dependent manner by 20 +/- 4, 28 +/- 5, 38 +/- 6, and 44 +/- 7 mm Hg in sham-operated rats and 30 +/- 3, 55 +/- 5, 78 +/- 5, and 113 +/- 8 mm Hg in rats with aortic coarctation. Indomethacin (1 microg/ml) or the endoperoxide/thromboxane blocker SQ29548 (1 microM) prevented AA renal vasoconstriction. Cyclooxygenase activity, cyclooxygenase-1 protein content, and mRNA expression were also increased in the renal tissue from the aortic coarctation rats compared with sham-operated rats. In conclusion, we suggest that during development of hypertension, the cyclooxygenase-1 mRNA is induced, and consequently cyclooxygenase-1 activity and AA metabolism are increased, resulting in augmented production of vasoconstrictor prostaglandins that mediate the potentiated responsiveness to AA or other vascular agonists that release AA, thus increasing peripheral vascular resistance. PMID- 11065218 TI - Electrophysiologic effects of lercanidipine on repolarizing potassium currents. AB - Blockade of cardiac repolarizing potassium channels by drugs may result in QT interval prolongation, eventually degenerating into "torsades de pointes," a life threatening arrhythmia. Lercanidipine (LER) is a recently introduced lipophilic calcium antagonist with no cardiodepressant activity and long-lasting antihypertensive action. Its chemical structure is characterized by the presence of a diphenylpropylaminoalkyl group, which is present in some of the drugs that have been reported to cause QT-interval prolongation. Our previous data demonstrated that LER blocks L-type calcium channels without affecting sodium current; however, no data are available concerning its effects on cardiac potassium channels. Transient outward (I(to)), delayed rectifier (I(K)), background currents, and action potential (AP) profile were measured from patch clamped ventricular myocytes isolated from rat, guinea pig, or human hearts using enzymatic dissociation procedures. LER did not affect I(K) (and I(Kr)) density and activation curve in guinea pig myocytes; the reversal potential of the background current (I(K1)) and its slope were not changed by the drug. Maximal diastolic potential (MDP) and duration of the AP measured at -60 mV (APD(-60)) were not significantly changed. I(to) density and activation curves measured in rat myocytes were similar in the absence and presence of 1 or 10 microM LER. Finally, the effect of LER was tested in human ventricular myocytes: superfusion with 1 microM LER did not affect MDP and APD(-60). I(to) density and the midpoint of activation and inactivation curves were similar in the absence and presence of LER. In conclusion, our data demonstrate that LER does not affect repolarizing potassium currents and action potential profile recorded from guinea pig, rat, and human ventricular myocytes. It is unlikely that LER could cause QT prolongation in vivo. PMID- 11065219 TI - Clevidipine blockade of L-type Ca2+ currents: steady-state and kinetic electrophysiological studies in guinea pig ventricular myocytes. AB - Steady-state and transient effects of clevidipine, a rapidly degraded dihydropyridine (DHP) L-type Ca2+ channel antagonist, were examined on I(Ca) in guinea pig ventricular myocytes. When myocytes were voltage-clamped with holding potential (V(H)) at -80 mV, 10 nM clevidipine decreased I(Ca) at 0 mV by approximately 30%, but >50% when V(H) was -40 mV. Rapid (<50 ms) perfusion switching and repeated depolarizations delivered at 0.5-2 Hz were used to determine the time constants of onset (tau(on)) and recovery from (tau(off)) clevidipine inhibition of I(Ca). The tau(on) and tau(off) were monoexponential functions of time. The tau(on) of I(Ca) inhibition decreased from 21.5 +/- 1.2 to 9.9 +/- 0.9 s when the rapidly applied [clevidipine] was increased from 10 to 100 nM at V(H) = -80 mV; tau(off) was independent of the applied [clevidipine] and was 23.9 +/- 1.1 s. The dissociation constant (K(D)) calculated for clevidipine at V(H) = -80 mV was 65 +/- 3 nM, similar to the IC50 of 78 nM determined in steady-state measurements. Decreasing V(H) to -40 mV increased tau(off) more than threefold to 81 +/- 6 s, and K(D) was markedly decreased to 9.0 +/- 0.8 nM (IC50, 7.1 nM at V(H) = -40 mV). The increased affinity at depolarized V(H) may contribute to the varying concentration-effect relation observed in vivo. PMID- 11065220 TI - Oxidative stress developed during open heart surgery induces apoptosis: reduction of apoptotic cell death by ebselen, a glutathione peroxidase mimic. AB - Apoptosis, a genetically controlled programmed cell death, has been found to play a role in ischemic reperfusion injury in several animal species including rats and rabbits. To examine whether this also is true for other animals, a surgically relevant model was established using an isolated in situ swine heart. Hearts were subjected to 15 min of normothermic regional ischemia by left anterior descending artery (LAD) occlusion followed by 30 min of normothermic cardioplegic arrest and 3 h of reperfusion. Oxygen free radicals have been shown to be the inducers of apoptosis and because reperfusion of ischemic myocardium is associated with the generation of free radicals, an additional group of hearts was preperfused with three different doses (5, 10, and 25 nM) ebselen, a glutathione peroxidase mimic, for 15 min before 15 min of LAD occlusion. Hearts were then subjected to 30 min of normothermic cardioplegic arrest followed by 3 h of reperfusion at normothermia. Control experiments were performed by perfusing the hearts for 4 h at normothermia. Two other groups of hearts were subjected to either 30 or 60 min of LAD occlusion followed by 30 min of cardioplegic arrest without subjecting them to reperfusion. At the end of each experiment, hearts were processed for the evaluation of apoptosis and DNA laddering. The in situ end-labeling (ISEL) technique was used to detect apoptotic cardiomyocyte nuclei while DNA laddering was evaluated by subjecting the DNA obtained from the cardiomyocytes to 1.8% agarose gel electrophoresis followed by photographing under UV illumination. The apoptotic cells appeared only after 90 min of reperfusion, as demonstrated by the intense fluorescence of the immunostained genomic DNA when observed under fluorescence microscopy. None of the ischemic hearts showed any evidence of apoptosis. These results were corroborated with the findings of DNA fragmentation showing increased ladders of DNA bands in the same reperfused hearts. The presence of apoptotic cells and DNA fragmentation in the myocardium was abolished by preperfusing the hearts in the presence of 10 nM ebselen, which also moderated the oxidative stress developed in the heart. Apoptotic cells and DNA ladders were completely absent in the hearts subjected to either 30 or 60 min of LAD occlusion. The results demonstrate that reperfusion of the ischemic heart induces apoptosis, which can be reduced with ebselen by reducing the oxidative stress associated with ischemia/reperfusion. PMID- 11065221 TI - Enhancement of the cAMP-induced apolipoprotein-mediated cellular lipid release by calmodulin inhibitors W7 and W5 from RAW 264 mouse macrophage cell line cells. AB - Apolipoprotein (apo) A-I generates high-density lipoprotein (HDL) by removing cellular cholesterol and phospholipid on the interaction with cells as a main source of plasma HDL. The reaction is induced by dibutylyl cyclic (dbc) adenosine monophosphate (AMP) in RAW 264, mouse macrophage cell line cells, and we investigated its pharmacologic modulation using this cell model. Release of cellular cholesterol and choline phospholipid by apoA-I was increased 9.9 and 4.2 times, respectively, by pretreatment of the cells with 300 microM dbcAMP for 24 h. Calmodulin inhibitors, W7 (N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide) and W5 (N(6-aminohexyl)-1-naphthalenesulfonamide), increased the apoA-I-mediated lipid release by 3 times from the dbcAMP-treated cells. The optimal drug concentrations (80 and 160 microM for W7 and W5, respectively) were not parallel with those reported for in vitro calmodulin inhibition (IC50, 28 and 240 microM for W7 and W5, respectively, toward phosphodiesterase activity), and in fact 40 microM W7 showed much stronger intracellular calmodulin inhibition than did 300 microM W5 using S7AAS2, a fluorescent peptide probe. Other calmodulin inhibitors such as amitriptyline, chlorpromazine, and trifluoperazine showed no effect on the apoA-I-mediated cholesterol release. In contrast to these results, neither dbcAMP nor W7 influenced the diffusion-mediated nonspecific cholesterol efflux to lipid microemulsion. We concluded that W7 and W5 increased the interaction of apoA-I with RAW 264 cells to generate more HDL. The effect did not seem directly correlated to their cal modulin inhibition or modulation of cAMP and protein kinase C. PMID- 11065222 TI - Short-term atorvastatin treatment improves endothelial function in hypercholesterolemic women. AB - Endothelial dysfunction represents the earliest stage of atherosclerosis and is usually present in hypercholesterolemia. Treatment with statins has been shown to normalize endothelial function in middle-aged men with hypercholesterolemia. We evaluated the effect over time of atorvastatin on the endothelial reactivity in postmenopausal hypercholesterolemic women (mean age, 58 +/- 6 years), receiving atorvastatin, 10 mg daily (n = 20) or American Heart Association step 1 diet (n = 10) for 8 weeks. Lipid profile and brachial artery flow-mediated vasodilation (FMV) were determined at baseline and after 1, 2, 4, and 8 weeks. FMV increased progressively in subjects treated with atorvastatin, and the difference was significant (p < 0.05 vs. baseline) after the second week (baseline 3.8 +/- 3%; first week, 4.8 +/- 3%; second week, 9.2 +/- 3%; fourth week, 11.0 +/- 3%; eighth week, 11.7 +/- 3%). No significant changes were observed in subjects receiving diet (baseline, 3.1 +/- 4%; first week, 2.4 +/- 2%; second week, 2.9 +/- 2%; fourth week, 3.1 +/- 2%; eighth week, 3.3 +/- 2%; p = NS). In the atorvastatin group, low-density lipoprotein (LDL) cholesterol showed a significant decrease since the first week (baseline, 228 +/- 37 mg/dl; first week, 171 +/- 32; second week, 147 +/- 27; fourth week, 139 +/- 29; eighth week, 135 +/- 27; all p < 0.05). In the control group, LDL cholesterol showed a smaller but significant (p < 0.05) reduction after the second week (baseline, 226 +/- 17 mg/dl; first week, 225 +/- 16; second week, 220 +/- 17; fourth week, 203 +/- 27; eighth week, 198 +/ 27). In conclusion, hypercholesterolemic women treated with atorvastatin show a significant improvement in endothelial reactivity after as early as 2 weeks of therapy. The extent to which these beneficial effects are attributable to cholesterol reduction or to a direct effect of the drug remains to be established. PMID- 11065223 TI - Comparison of endothelium-dependent relaxation in carotid arteries from Japanese white and Watanabe heritable hyperlipidemic rabbits. AB - Modifications by atherosclerosis of endothelium-dependent and -independent relaxations were evaluated in carotid arteries isolated from Watanabe heritable hyperlipidemic (WHHL; age 20-29 months) and age-matched Japanese white (JW) rabbits. Marked, patchy atherosclerotic lesions were observed in all WHHL rabbit arteries. Endothelium-dependent relaxations induced by acetylcholine, partly depressed by N(G)-nitro-L-arginine (L-NA), were significantly inhibited in the WHHL rabbit arteries with atherosclerosis, compared with those in the arteries without atherosclerotic lesions from JW and WHHL rabbits. No difference was observed in the relaxation caused by superoxide dismutase in these arteries. Conversely, endothelium-dependent relaxations by substance P were greater in the arteries with and without atherosclerosis from WHHL rabbits than in the arteries from JW rabbits. Endothelium-independent relaxations elicited by sodium nitroprusside and 2,2-(hydroxynitrosohydrazino)bis-ethanamine (NOC18) did not differ in the arteries from JW and WHHL rabbits. The responses to acetylcholine and substance P of JW rabbit arteries with the endothelium were not attenuated by treatment with pertussis toxin. L-NA-resistant, endothelium-dependent relaxations by substance P were almost abolished by charybdotoxin, and atherosclerosis did not alter the response. It is concluded that endothelial functions, evaluated by substance P, in rabbit carotid arteries are not impaired by atherosclerosis and by long exposure to hyperlipidemia in vivo. Dysfunction of muscarinic receptors may be involved in the depressed response to acetylcholine. As far as the arteries used in the present study are concerned, responses mediated possibly by endothelium-derived hyperpolarizing factor (EDHF) are unlikely to be modulated by atherosclerosis. PMID- 11065224 TI - Modulation of contractions to ergonovine and methylergonovine by nitric oxide and thromboxane A2 in the human coronary artery. AB - This study explored the modulatory effects of nitric oxide and thromboxane A2 on contractions to ergonovine and methylergonovine in human coronary arteries. To elucidate the different role of nitric oxide synthase in the response to the ergot alkaloids, the serotonin (5-HT) receptors involved in nitric oxide synthase in the response to the ergot alkaloids, the 5-HT receptors involved in nitric oxide release and the contraction of the vascular smooth muscle were characterized with more selective 5-HT-receptor agonists and antagonists. Rings of human coronary arteries from explanted hearts were suspended in organ chambers for isometric tension recording. After testing for contractile (potassium chloride, 60 mM) and endothelial function (substance P, 10(-8) M), respectively, they were exposed to ergot alkaloids or other agonists in the absence or presence of U 46619 (10(-9) M), or nitro-L-arginine (10(-4) M), or both. Ergonovine and methylergonovine were comparable, weak vasoconstrictors in untreated preparations. Contractions to ergonovine were augmented by U 46619, but not by nitro-L-arginine. Contractions to methylergonovine were augmented only by combining U 46619 and nitro-L-arginine. Serotonin and methylergonovine, but not ergonovine, elicited endothelium-dependent, nitric oxide-mediated relaxations. Nonselective 5-HT(1B/1D)-receptor stimulation caused both contractions and relaxations; selective 5-HT1B stimulation caused relaxations only. In the human coronary artery, contractions to ergonovine are not dependent on NO release but are synergistically augmented by thromboxane. Methylergonovine causes similar effects on the vascular smooth muscle, but contractions are inhibited by the release of NO from the endothelium. The 5-HT receptor on the endothelium appears to be different from the receptor on the vascular smooth muscle, which mediates the contractile response to the ergot alkaloids. PMID- 11065225 TI - Reproducibility of laser Doppler imaging of skin blood flow as a tool to assess endothelial function. AB - Endothelial dysfunction might be an important and early event in the pathogenesis of major cardiovascular diseases. Therefore, the evaluation of endothelial function in humans may be of great clinical relevance. Usual methods for that purpose are either invasive and/or technically demanding. In the dermal microcirculation, endothelial function may be assessed noninvasively from the laser Doppler measurement of increases in blood flow after either the transdermal application of acetylcholine by iontophoresis, or the release of transient arterial occlusion (reactive hyperemia). An endothelium-independent response may be provided by the iontophoresis of sodium nitroprusside. This approach is notable for technical simplicity, but of uncertain reproducibility. Sixteen young, healthy, nonsmoking males were examined in the fasting state. Changes in skin blood flow were measured with a laser Doppler imager during the iontophoresis of acetylcholine and sodium nitroprusside, as well as during reactive hyperemia, on two different days, at each of two different sites on the volar face of the forearm. Nonspecific effects related to the stimulation of terminal nerve fibers by the iontophoretic current were suppressed by prior surface anesthesia. The iontophoresis of acetylcholine and sodium nitroprusside induced a seven- to eightfold increase in dermal blood flow. The corresponding figure for peak reactive hyperemia was approximately fourfold. The mean coefficients of variation of responses recorded on different days, on the same site, in the same individual were <10% for iontophoresis of acetylcholine and for peak reactive hyperemia, and between 10 and 20% for iontophoresis of sodium nitroprusside. This day-to-day variation was significantly smaller than the site to-site variation (p < 0.01 for all three responses). Endothelium-dependent and independent responses of dermal blood flow evaluated with laser Doppler imaging are highly reproducible from day to day, at least in healthy nonsmoking young male subjects, and provided some simple precautions are observed, foremost among which is the strict standardization of the recording site. These observations may have implications for the testing of endothelial function in clinical studies. PMID- 11065226 TI - IV. Anticoagulant activity of compound 48/80: inhibition of factor VII activation in leukemia THP-1 monocytes. AB - Our previous study described a novel biologic function of compound 48/80 (48/80) in the downregulation of monocytic tissue factor (TF)-initiated hypercoagulation in response to bacterial endotoxin (lipopolysaccharide; LPS). The inhibition was not due to the blockade of LPS cell signaling, as evidenced by the unaffected LPS induced TF synthesis. We herein determined the mechanism by which 48/80 inhibits the extrinsic coagulation in agonist-challenged THP-1 monocytes. LPS as well as A23187 substantially induced TF activity. TF synthesis was enhanced by LPS but not by A23187. However, the elevated FVII binding to monocytes accompanying the upregulation of factor VII (FVII) activation was uniformly observed in both cases. A 5-min preincubation of the cells with a sheep anti-humanTF antibody (anti-hTF Ab) showed the downregulation of FVII activation, indicating a regulatory role of FVII binding in the modulation of the extrinsic coagulation. The 48/80 blocked FVII binding to monocytes, leading to the preferential inhibition of FVII activation. As the result of the diminished FVIIa formation, monocytic TF-initiated extrinsic coagulation was downregulated in agonist challenged THP-1 monocytes. PMID- 11065227 TI - Cardiac ischemia oxidizes regulatory thiols on ryanodine receptors: captopril acts as a reducing agent to improve Ca2+ uptake by ischemic sarcoplasmic reticulum. AB - We tested the hypothesis that ischemia alters sarcoplasmic reticulum (SR) Ca2+ transport by oxidizing regulatory thiols on ryanodine receptors (RyRs), and that membrane-permeable sulfhydryl-containing angiotensin-converting enzyme (ACE) inhibitors protect against ischemia-induced oxidation and explain in part, the therapeutic actions of captopril. Ca2+ uptake and adenosine triphosphatase (ATPase) activity was measured from SR vesicles isolated from control or ischemic dog and human ventricles and compared with or without sulfhydryl reductants. The rate and amount of Ca2+ uptake was lower for canine ischemic SR compared with control (6.5 +/- 0.2 --> 18.5 +/- 1.1 nmol Ca2+/mg/min and 123.1 +/- 4.7 --> 235.0 +/- 17.3 nmol Ca2+/mg; n = 8 each). Captopril, dithiothreitol (DTT), glutathione (GSH), and L-cysteine increased the rate and amount of Ca2+ uptake by canine and human ischemic SR vesicles by approximately 50%. Reducing agents had no effect on Ca2+- ATPase activity in either canine control or ischemic (approximately 40% less than control) SR. Captopril was as potent as DTT at reversing the oxidation of skeletal and cardiac RyRs induced by reactive disulfides (RDSs) or nitric oxide (NO). In neonatal rat myocytes, RDSs or NO triggered SR Ca2+ release and increased cytosolic Ca2+, an effect reversed by captopril and DTT but not GSH or cysteine. Pretreatment of myocytes with captopril (exposure and then wash) inhibited Ca2+ elevation elicited by RDSs or NO, indicating that captopril is an effective, membrane-permeable intracellular reducing agent. Thus, net SR Ca2+ accumulation is reduced by ischemia in part due to the oxidation of thiols that gate RyRs, an effect reversed by captopril. PMID- 11065228 TI - Effects of prolonged caffeine consumption on cardiac contractile function in rats. AB - The purpose of the study was to explore effects of prolonged caffeine administration on the contractile function and myocardial energy metabolites of the isolated rat heart. Caffeine treatment for 1 week (10 mg/kg, i.p., twice a day) was followed by unchanged pump function of the isolated heart, but reduced maximal left ventricular (LV) systolic pressure by 14% (p < 0.05). Caffeine consumption during 8-9 weeks (0.1% water solution) was also followed by unchanged maximal pump function but increased maximal double product (LV developed pressure multiplied by heart rate) by 23% (p < 0.05). The hearts of caffeine-consumed rats also maintained a higher level of the pump function at a high rate of atrial electrostimulation. The myocardial content of adenosine triphosphate (ATP), creatine phosphate, as well as creatine was slightly but insignificantly increased after caffeine consumption. Results show that in the course of prolonged caffeine treatment, the maximal myocardial contractile function first decreases and then increases, showing adaptation of the heart. PMID- 11065229 TI - Thapsigargin induces apoptosis in cultured human aortic smooth muscle cells. AB - Vascular remodeling is a key feature of many pathologic states, including atherosclerosis, or hypertension. Vascular smooth muscle cells participate in determining the vessel structure by several mechanisms such as cell migration, cell growth, or cell death (necrosis or apoptosis). Here we report that thapsigargin, an inhibitor of endoplasmic reticulum Ca2+ -adenosine triphosphatase (ATPase), is able to induce apoptosis in human vascular smooth muscle cells (HVSMCs). Apoptosis was assessed by three different methods: differential chromatin binding dye staining. cytoplasmic histone-associated DNA fragments detection by enzyme-linked immunosorbent assay (ELISA) and terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). When HVSMCs were treated for 1 h with thapsigargin (100 nM-10 microM), there was a concentration-dependent increase in both parameters 24 h after the thapsigargin pulse. When a time-course experiment was performed, both parameters were significantly enhanced from 3 to 6 h after the exposure to thapsigargin. We conclude that thapsigargin promotes apoptosis in HVSMCs, providing a useful tool for the study of programmed cell death in human vascular smooth muscle. PMID- 11065230 TI - The effect of adenosine on blood pressure variability in sinoaortic denervated rats is mediated by adenosine A2a-Receptor. AB - It is known that adenosine decreases blood pressure (BP) level as well as blood pressure variability (BPV). However, there is little information about the effect of adenosine on BPV. With a computerized analytic system for BP and heart rate (HR) that could sample the data continuously in conscious, freely moving rats, we studied the effects of different agonists and antagonists of adenosine receptors on BPV in sinoaortic denervated (SAD) rats. It was found that both adenosine and 5'-N-cyclopropyl-carboxamidoadenosine (CPCA, a selective adenosine A,-receptor agonist) decreased BPV. whereas N6-cyclopentyladenosine (CPA, a selective adenosine A1-receptor agonist) had no significant effect on BPV. When the rats were pretreated with theophylline (the nonselective adenosine-receptor antagonist), the inhibitory effects of adenosine as well as CPCA on BPV were abolished. Furthermore, it was found that 8-(3-chlorostyryl)caffeine (CSC, a selective adenosine A2a-receptor antagonist), also could prevent such an effect on BPV of CPCA. By itself, however, neither theophylline nor CSC had any influence on BPV. These results suggest that the effect of adenosine on BPV is mediated by adenosine A2a-receptor. PMID- 11065231 TI - Management of septic patients without the consultation services of infectious disease specialists. PMID- 11065232 TI - Expression of c-kit in gastrointestinal stromal tumors. PMID- 11065233 TI - Does brain damage (hepatic encephalopathy) improve without any complication and sequellae after liver transplantation? PMID- 11065234 TI - Thrombin and antithrombin in Binswanger's disease. PMID- 11065235 TI - Infected aneurysms. PMID- 11065236 TI - The effects of long-acting nitrates on 5-year cardiac events of patients with coronary thrombolytic therapy for acute myocardial infarction. AB - OBJECTIVE: A retrospective study was performed to evaluate the effects of using long-term long acting nitrates without a dose-free interval in treating patients undergoing thrombolytic therapy for acute myocardial infarction (MI). PATIENTS AND METHODS: A total of 297 patients taking prescribed medication for secondary prevention of the MI were selected for the study. They were divided into a nitrate group consisting of 222 patients who had continuously received long acting nitrates without a dose-free interval, and a control group consisting of 75 patients who were not able to use the long-acting nitrates because of adverse effects. The primary endpoint was cardiac events, either cardiac death or a nonfatal MI, in five years. RESULTS: The incidence of primary endpoint in five years was 13.4 percent in the nitrate group and 6.2 percent in the control group, a 2.2-fold increase in risk. However, the difference was not significant. After adjustment for age, there was no statistically significant difference between the incidence of primary endpoint in the nitrate group (9.8%) and that in the control group (5.7%). A Cox proportional-hazards regression analysis revealed that the long-acting nitrates were not related to the incidence of primary endpoint (p=0.23). CONCLUSION: The administration of long-term long-acting nitrates without a dose-free interval had no benefit of reducing the incidence of cardiac events of patients undergoing thrombolytic therapy for acute MI. PMID- 11065237 TI - Psychobehavioral and immunological characteristics of HTLV-1 carriers and non carriers with persistently low natural killer cell activity. AB - OBJECTIVE: To clarify the differences in immunological and psychobehavioral characteristics of HTLV-1 carriers and non-carriers with persistently low natural killer (NK) cell activity. METHODS: The individuals with persistently low NK cell activity were divided into HTLV-1 carriers and non-carriers. NK cell activity, lymphocytic proliferation, lymphocyte subsets (CD4+, CD8+, CD16+, CD20+, CD56+), and psychobehavioral responses were examined. PATIENTS: Of 296 outpatients with physical complaints, 30 patients with persistently low NK cell activity (10 HTLV 1 carriers and 20 HTLV-1 non-carriers) and 20 healthy controls negative for HTLV 1 antibody and with normal NK cell activity were randomly selected. RESULTS: In HTLV-1 carriers with persistently low NK cell activity, no significant differences were observed in NK cell subsets (CD16+ and CD56+) and psychobehavioral responses compared with the healthy controls. In HTLV-1 non carriers, NK cell subsets were significantly low, and depression, anxiety and fatigue were significantly greater than in healthy controls. CONCLUSIONS: These findings suggest that persistently e low NK cell activity in HTLV-1 carriers might be reduced due to the HTLV-1 infection. On the other hand, the reduction in the NK cell activity in HTLV-1 non-carriers appears a to be related to depression, anxiety, and fatigue. PMID- 11065238 TI - Cerebral blood flow and cessation of cigarette smoking in healthy volunteers. AB - OBJECTIVE: The mechanisms by which cessation of cigarette smoking may improve regional cerebral blood flow (rCBF) and the details of the possible relationship remain unclear. Xenon-133 inhalation was used to determine rCBF in six smokers at baseline (during smoking) and again 6 and 9 years after they had quit smoking (quitter group). A control group of eight nonsmokers (nonsmokers group) underwent similar serial determinations. RESULTS: Regional CBF in quitters had decreased significantly after the 6 years, but improved significantly after 9 years, when abstinence had been maintained from 4 to 6 years. In the nonsmokers group rCBF did not change significantly over 9 years. CONCLUSION: Cessation of cigarette smoking improves cerebral circulation, but this effect requires several years. PMID- 11065239 TI - Nationwide survey of the annual prevalence of viral and other neurological infections in Japanese inpatients. AB - OBJECTIVE: To estimate the annual prevalence of viral and other neurological infections at large hospitals in Japan during the period from 1989 to 1991. METHODS: A nationwide questionnaire survey on the numbers of inpatients with viral and other neurological infections was sent for completion to the chiefs of Departments of Internal Medicine, Neurology and Pediatrics at all hospitals with more than 200 beds. RESULTS: The average annual number of inpatients (and the number per 10(6) population) with encephalitis in large hospitals was estimated to be 2,200+/-400 (17.7+/-3.2), while it was 32,000+/-16,000 (258+/-129) for meningitis, and 650+/-50 (5.2+/-0.4) for myelitis. Among the inpatients with encephalitis, meningitis, and myelitis, an unknown etiology was the most common (51.2% in encephalitis, 73.2% in meningitis, and 36.3% in myelitis), followed by a viral etiology for all three diseases. CONCLUSION: The first estimate was made of the annual prevalence of viral and other neurological infections and their etiology in Japan. PMID- 11065240 TI - Current practice of management of bacteremic sepsis: a study in a tertiary care teaching hospital in Japan. AB - OBJECTIVE: To investigate how patients with bacteremic sepsis are managed in a tertiary care teaching hospital. PATIENTS AND METHODS: Prospective observational study on patients with bacteremic sepsis. Clinical and microbiological characteristics of bacteremic sepsis were analyzed in relation to prognosis. Severity of the illness was quantitatively analyzed by the APACHE (Acute Physiology, Age, Chronic Health Evaluation) III scoring system. Also investigated was how closely physicians paid attention to acute physiological alterations in patients. RESULTS: The 28-day mortalities in fifty hemodynamically stable patients and in twenty-three septic shock patients were 26% and 52%, respectively (p=0.028). Gram-positive organisms accounted for 54% of all organisms, with the mortality and incidence of septic shock being the same as with Gram-negative infections. The mean APACHE III score was 42.9 in survivors, and 76.5 in non survivors (p < 0.001). Although serum levels of C-reactive protein and acute physiology score (APS) was significantly higher in non-survivors than in survivors, the correlation with APACHE III score was more prominent in APS. The number of vital signs recorded was 1.67 in physicians and 3.6 in nurses (p < 0.001). CONCLUSIONS: The present study proved that the APACHE III score accurately discriminates between survivors and non-survivors of patients with sepsis. By addressing the need for an objective evaluation of severity of illness, it strongly recommends that physicians should be made aware of physiologically defined sepsis and that they should pay closer attention to patients' physiological alterations to identify the development of sepsis in critically ill patients. PMID- 11065241 TI - Mixed carcinoid-adenocarcinoma of the liver. AB - A 61-year-old man with a mixed carcinoid-adenocarcinoma of the liver is described. Microscopic examination of the lesion showed a differentiated adenocarcinoma with distinct carcinoid components that stained positively for argyrophil. The tumor cells contained serotonin granules on immunohistochemical studies. Detailed examination disclosed no primary tumor in the gastrointestinal tract or in any other organ. Resection was considered impractical because there were multiple tumors. The patient received chemotherapy six times (cisplatin 60 mg/m2, epirubicin 40 mg/ m2 per month). The multiple tumors gradually shrank. At the time of this writing, the patient is still alive. To our knowledge, this is the first reported case of mixed carcinoid-adenocarcinoma of the liver. PMID- 11065242 TI - Gastrointestinal stromal tumors of the small intestine that expressed c-kit protein. AB - We report two patients with gastrointestinal stromal tumors (GISTs) of the small intestine that expressed c-kit protein (CD117). One was a 68-year-old woman with epigastralgia and vomiting. A submucosal tumor of the upper jejunum was detected, and partial resection was carried out. The histology revealed a GIST negative for CD34 but positive for CD117. The other was a 42-year-old woman with progressive anemia, melena and lower abdominal pain. Intussusception was detected, and a partial resection was carried out. A submucosal tumor of the lower jejunum was noted. The histology revealed a GIST positive for both CD34 and CD117. PMID- 11065243 TI - Malignant peritoneal mesothelioma associated with deep vein thrombosis following radiotherapy for seminoma of the testis. AB - A 52-year-old man developed malignant peritoneal mesothelioma 17 years after radiotherapy for seminoma of the testis. Although asbestos exposure is considered to be the major risk factor for the development of malignant mesothelioma, prior therapeutic radiation has also been postulated as a causative factor. The unexplained appearance of ascites or pleural effusion within a previously irradiated area should be considered suggestive of malignant mesothelioma in any long-term survivor of cancer. In addition, the patient suffered a deep vein thrombosis four years before the diagnosis of mesothelioma. Deep vein thrombosis is a common complication of malignant disease, and is often the first clue to occult malignancy. PMID- 11065244 TI - Reversibility of serum NH3 level in a case of sudden onset and rapidly progressive case of type 2 citrullinemia. AB - A 48-year-old male presented with an acute change in mental status due to a marked elevation of plasma NH3 and was diagnosed with citrullinemia with amino acid analysis of blood. Hemodialysis and hemodiafiltration were performed, but serum chemical analysis did not show any improvement which led us to terminate dialysis following intensive care for 3 days. Surprisingly, NH3 level had decreased by 6 days after admission, coinciding with normalization of the size of the pupils. Since spontaneous remission had never been discussed, we discuss this relatively rare, but clinically significant entity with regard to its acute phase management and its potential reversibility. PMID- 11065245 TI - Congenital hepatic fibrosis with fatal cholestatic liver damage. AB - A 23-year-old male with congenital hepatic fibrosis died because of progressive cholestatic liver damage. Pathologically, marked extension of fine fibers along the sinusoids in addition to fibrosis in Glisson's sheath, miniaturization and pseudo-glandular formation of hepatocytes as parenchymal damage, and nodular regenerative hyperplasia were considered as the cause of rapid aggravation of liver damage or portal hypertension. PMID- 11065246 TI - Esophageal perforation associated with profound shock successfully managed with hemodynamic assistance using percutaneous cardiopulmonary support. AB - A 51-year-old man was admitted to our hospital with complaints of severe chest pain, nausea, and vomiting. These symptoms had progressed rapidly and he was in shock. It was necessary to make a correct diagnosis as early as possible. However, the hemodynamic condition of the patient deteriorated rapidly before a definitive diagnosis could be established in spite of conventional therapies. Under hemodynamic assistance with percutaneous cardiopulmonary support (PCPS), a final diagnosis of esophageal perforation was made by esophagography. Our report illustrates a new application of PCPS for highly selected cases of noncardiogenic shock as a "bridge" until an accurate diagnosis is made and a specific treatment is applied. PMID- 11065247 TI - Acute aortic valvular regurgitation secondary to avulsion of aortic valve commissure in a patient with pseudoxanthoma elasticum. AB - A 68-year-old woman developed acute pulmonary edema due to severe acute aortic valvular regurgitation. At the time of emergency surgery, it turned out to result from spontaneous avulsion of the aortic valve commissure. Later, the patient was diagnosed to have pseudoxanthoma elasticum based on typical skin lesions. Connective tissue abnormalities associated with pseudoxanthoma elasticum might have contributed to the development of the avulsion of the aortic valve in this particular patient. PMID- 11065248 TI - Slowly progressive insulin-dependent diabetes mellitus in an elderly patient with Graves' disease. AB - We report a case of slowly progressive insulin-dependent diabetes mellitus in an elderly patient with Graves' disease. A 69-year-old man presented with apathetic thyrotoxicosis and weight loss. Laboratory findings indicated insulin-dependent diabetes mellitus (IDDM) with Graves' disease. Human leukocyte antigens DR4 and DR9, which are recognized as markers for IDDM with autoimmune thyroid disease, were detected. The clinical course of the IDDM was compatible with the slowly progressive type. Onset of this disease during old age is rare, and such cases should be analyzed with a thyroid function test because the symptom of thyrotoxicosis may be masked in the elderly. PMID- 11065249 TI - Occupational asthma due to exposure to African cherry (Makore) wood dust. AB - A 35-year-old man who had been a carpenter and a cabinet worker for over 15 years, was referred to our clinic with a 4-month history of cough, chest tightness and difficulty in breathing which occurred within minutes of exposure to African cherry wood (Makore). He developed a dual asthmatic reaction on specific challenge test with an extract of African cherry wood dust. Thus, the diagnosis of occupational asthma due to exposure to African cherry wood dust was confirmed by the specific challenge test. The mechanism of asthma due to African cherry wood dust exposure is not clear. PMID- 11065250 TI - Follicular dendritic cell sarcoma: ultrastructural and immunohistochemical studies. AB - A rare case of follicular dendritic cell (FDC) sarcoma is reported. A 71-year-old woman was admitted for evaluation of constipation. Computerized tomography showed cervical, supraclavicular, retroperitoneal, and paraaortic lymphadenopathies. Histological findings from a cervical lymph node revealed Hodgkin's disease at first. But tumors that arose both in the cervical and the left interscapular regions during the chemotherapy were immunohistochemically confirmed to be of follicular dendritic cell origin. The ultrastructural findings were consistent with those of FDC sarcoma. FDC sarcoma is a rare nonlymphoid cell-derived malignant tumor originating from the lymphoid tissue. The diagnosis of FDC sarcoma is most accurately established by immunohistochemical methods, using its specific markers. PMID- 11065251 TI - Slowly progressive dystonia following central pontine and extrapontine myelinolysis. AB - A 28-year-old woman was hospitalized with dysarthria and oro-mandibular and upper limb dystonia. Approximately 8 years prior to the current admission, the woman became severely hyponatremic due to traumatic subarachnoid hemorrhage-related SIADH. Brain MRIs showed a signal increase in the central pons, thalamus and striatum on T2 weighted images compatible with central pontine and extrapontine myelinolysis. From a few months after that event, dystonia progressed slowly over the subsequent 8 years. We speculate that the particular damage chiefly to the myelin structures by myelinolytic process may have caused an extremely slow plastic reorganization of the neural structures, giving rise to progressive dystonia. PMID- 11065252 TI - Bilirubin adsorption therapy and subsequent liver transplantation cured severe bilirubin encephalopathy in a long-term survival patient with Crigler-Najjar disease type I. AB - Crigler-Najjar disease (CN) type I is characterized by persistent unconjugated hyperbilirubinemia from birth. The male patient here was diagnosed with this disease as a neonate and had been treated by phototherapy. At age 16 he suddenly developed generalized convulsions, followed by impaired cognitive function. The serum level of bilirubin was extremely high (total bilirubin: 41.7 mg/dl) and there were no other detectable causes responsible for the metabolic encephalopathy. He received bilirubin adsorption therapy several times, and the bilirubin encephalopathy improved in response to the fall in the serum level of bilirubin. After this he underwent a successful liver transplantation in Australia, and recovery of his mental faculties was satisfactory. Within the subsequent 3 years epileptic abnormal discharges on the electroencephalogram disappeared. Phototherapy alone can not prevent the rise in the serum level of bilirubin in adolescent or adult patients with CN type I, therefore such patients tend to experience life-threatening bilirubin encephalopathy. To save patients with the acute onset type of bilirubin encephalopathy, sufficient bilirubin adsorption followed by liver transplantation appears to be the most recommended therapeutic approach. PMID- 11065253 TI - Effects of an antithrombin drug in patients with subacute exacerbations of Binswanger disease. AB - The blood coagulation system has been shown to be activated in subacute exacerbations of Binswanger disease (BD). In our previous study, the antithrombin drug argatroban t ameliorated the neurological exacerbations in a BD patient with antiphospholipid antibody syndrome. We have further examined the therapeutic efficacy of argatroban in 3 BD patients with subacute exacerbations, but without any immune-mediated prothrombotic complications. In 1 out of these 4 patients, treatment with sodium ozagrel, an antiplatelet drug was applied, but was ineffective. In all patients, argatroban treatment reduced the levels of the hemostatic markers, with a corresponding improvement in cognitive dysfunction and gait disorders. These results suggest that the antithrombin effect is true also for BD patients not compromised by the immune-mediated prothrombotic condition. PMID- 11065254 TI - Interstitial pneumonia complicated by Sjogren's syndrome, Hashimoto's disease, rheumatoid arthritis and primary biliary cirrhosis. AB - A 66-year-old woman diagnosed as having Hashimoto's disease and rheumatoid arthritis manifested interstitial pneumonia. We diagnosed Sjogren's syndrome and primary biliary cirrhosis as complications in this case. Steroid therapy was relatively effective for the interstitial pneumonia which was in an active state; however, during tapering of the steroid, there was a relapse and also severe dry throat. Cyclophosphamide was added and was effective in the prevention of recurrence. Even after discontinuation of steroid therapy, her general condition is stabilized. It is very important to carefully investigate other organ involvement as a prognostic factor in cases in which there are multiple autoimmune diseases. PMID- 11065255 TI - Churg-Strauss Syndrome with pleural involvement. AB - A 51-year-old Japanese man with Churg-Strauss Syndrome (CSS) diagnosed by pleural biopsy is described. He was hospitalized because of high fever and bilateral knee, elbow and shoulder joint pain. Chest roentgenogram and chest computed tomography (CT) scan revealed bilateral massive pleural effusion. Pleural biopsy revealed eosinophilic infiltration and necrotizing granulomas. He was treated with oral prednisolone and his symptoms improved. This is the first report of CSS diagnosed by pleural biopsy. PMID- 11065256 TI - Type II diabetes mellitus and primary Sjogren's syndrome complicated by pleural effusion. AB - A 73-year-old man was admitted to our hospital because of pleural effusion and nephrotic syndrome. Sjogren's syndrome (Sjs) was diagnosed based on a positive test for antibodies to Ro and La, and the result of labial salivary gland biopsy. The pleural effusion showed a high number of lymphocytes and high titers of antibodies to Ro and La. By immunohistochemistry, it was determined that infiltrating CD3+ cells predominated over infiltrating CD20+ cells in the pleura. Nephrotic syndrome was also present, which, as confirmed by renal biopsy was due to advanced diabetic nephropathy. Here, we report a case of Type II diabetes mellitus and primary Sjs complicated by pleural effusion, discuss the available treatment for pleural effusion. PMID- 11065257 TI - Churg-Strauss syndrome associated with pregnancy. AB - Churg-Strauss syndrome, an uncommon condition, occurs even more rarely in association with pregnancy. One month postpartum, a 25-year-old Japanese woman was hospitalized for dyspnea, fever, diarrhea, and complaints of dysesthesia in the right lower limb and right lumbar and abdominal region. Marked eosinophilia was present. Symptoms of bronchial asthma had developed early in the pregnancy, which concluded with delivery of a healthy baby at 39 weeks of gestation. Churg Strauss syndrome was diagnosed and prednisolone was administered with a good response leading to remission. Three years after the first pregnancy, low-dose steroid therapy was continued through another pregnancy. The patient delivered another healthy baby at 39 weeks of gestation, this time with no exacerbation of symptoms. PMID- 11065258 TI - A diabetic patient with scrotal subcutaneous abscess. AB - A 51-year-old type 2 diabetic patient with a scrotal subcutaneous abscess is reported. He was diagnosed as having diabetes mellitus five years earlier. He had left scrotal swelling and pain with granulocytosis, elevated C-reactive protein and hyperglycemia. He was successfully treated with incision and drainage (Streptococcus agalactiae was identified in the pus), debridement, antibiotics, immunoglobulin and insulin. This case resembled Fournier's gangrene, an infective necrotizing fasciitis of the perineal, genital or perianal regions. Diabetes mellitus is a basic disorder often associated with Fournier's gangrene. Scrotal subcutaneous abscess should be prevented from progressing to Fournier's gangrene with early and appropriate treatment. PMID- 11065259 TI - Hepatic artery pseudoaneurysms in a patient treated for miliary tuberculosis. AB - A 70-year-old woman with fever was admitted to our hospital. She was diagnosed as miliary tuberculosis and treated with antituberculous drugs. After seven weeks of therapy, she developed a sudden sharp upper abdominal pain and shock. Angiography of the celiac artery showed two hepatic artery pseudoaneurysms with extravasation. The hemorrhage was successfully stopped by microcoil embolization. The clinical course suggested that miliary tuberculosis had caused the pseudoaneurysms. Although aneurysms rarely occur as a complication of miliary tuberculosis, they should be diagnosed as early as possible because of the high rate of rupture and associated high mortality rate. PMID- 11065260 TI - Common bile duct measurements in an elderly population. AB - We prospectively evaluated the diameter of the common bile duct in 1,018 patients between the ages of 60 to 96 over a 4 year period to determine if there is a significant change in its size with aging. All of the patients included in the study were being evaluated primarily for carotid or peripheral vascular disease. Any patients with a history of biliary disease (i.e., bilirubin level greater than 1.5 mg/ml, cholecystectomy, or cholelithiasis) were excluded. Ultrasonography of the common bile duct was performed only in those patients with no subjective abdominal pain or icterus. Our results demonstrated a small although statistically significant increase in the caliber of the common bile duct with increasing age (60 years old or less, mean diameter 3.6 mm +/- 0.2mm, versus over 85 years old, mean diameter 4 mm +/- 0.2 mm, P = 0.009). Although the common bile duct did increase in size with aging, 98% of all ducts remained below 6 to 7 mm, the commonly accepted upper range of normal. PMID- 11065261 TI - Sonographic evaluation of the thenar compartment musculature. AB - The thenar region was studied with ultrasonography in 10 healthy volunteers. All thenar muscles could be identified and their course followed entirely. In addition, their function could be assessed by scanning during unresisted or resisted active movements. Standard approach, normal appearance, and dynamic tests for each muscle are described. PMID- 11065262 TI - Sonographic evaluation of cellulitis in children. AB - Within a 3 year period from July 1996 to July 1999, a total of 105 children with clinical diagnosis of cellulitis were evaluated by ultrasonography. Eighty-six children (age range, 17 days to 15 years) fulfilling the sonographic criteria for diagnosis of cellulitis were enrolled into the study. The sonographic features were used to correlate with clinical symptoms and their duration, the peripheral leukocyte count, and the serum C-reactive protein level. Pus aspiration for immediate microscopic and later bacteriologic studies was carried out under sonographic guidance. Ultrasonographic features of cellulitis included subcutaneous tissue thickening without distortion and pus (25 cases, 29%), distortion of subcutaneous tissue without pus accumulation (26 cases, 30%), distortion of subcutaneous tissue with pus accumulation (19 cases, 23%), and distortion of tissue with abscess formation (16 cases, 18%). The presence of sonographic features of tissue distortion with or without pus accumulation, including abscess formation in children with cellulitis, correlated with a longer duration of symptoms (greater than 4 days), the presence of high-grade fever, higher peripheral leukocyte count, and higher serum C-reactive protein levels. Those patients who underwent sonographically guided aspiration or surgical intervention showed a shorter hospital stay and fever duration than those without such aspiration. Our results indicated that ultrasonography is of great value in managing cellulitis by providing information regarding the progression of inflammation. Sonographically guided aspiration of pus may be a treatment of choice, as it may decrease the need for operation. PMID- 11065263 TI - Evaluation of flash echo imaging of the canine gastrointestinal tract. AB - Although it is important to assess gastrointestinal blood flow, no generally useful, noninvasive assessment method has been established. Harmonic flash echo imaging, which is an intermittent second harmonic imaging technique, has recently become available to evaluate blood flow. We investigated the usefulness of harmonic flash echo imaging in the assessment of the gastrointestinal tract, and we used this technique to study the effect of nicotine on small bowel blood flow. Harmonic flash echo imaging was performed at the beginning of intravenous injection of a contrast agent. It was also performed on the small bowel immediately before and 10 min after nicotine administration to evaluate blood flow. Gastric and small bowel walls were clearly enhanced on the primary images. Small bowel enhancement, which is regarded as transmural blood flow, significantly decreased after nicotine administration. Harmonic flash echo imaging appears to be useful in the assessment of the transmural blood flow in the gastrointestinal wall. PMID- 11065264 TI - Sonographic evaluation of the pancreatic duct in normal children and children with pancreatitis. AB - We investigated the diameter of pancreatic duct using ultrasonography in 51 children with pancreatitis and age-matched healthy control children over a 5 year period. The diameters of pancreatic duct and pancreatic body were measured simultaneously by sonography. The mean ages of children with acute pancreatitis and chronic pancreatitis were 9.7 +/- 3.9 and 10.3 +/- 3.1 years, respectively (range, 1 to 8 years). The mean age of normal children was 9.6 +/- 5.3 years. A significant difference was found in diameter of the pancreatic duct between children with acute and chronic pancreatitis versus that of age-matched control. In addition, a significant difference in diameter of the pancreatic body was found between children with acute pancreatitis and age-matched controls, but there was no marked difference in diameter of the pancreatic body between normal persons and those with chronic pancreatitis. The mean diameters of the pancreatic duct in acute pancreatitis and chronic pancreatitis were 2.34 +/- 0.47 mm and 2.84 +/- 0.67 mm, respectively, which was greater than that of normal children (1.65 +/- 0.45 mm). Pancreatic ducts with diameters greater than 1.5 mm in children between 1 and 6 years, greater than 1.9 mm at ages 7 to 12 years, or greater than 2.2 mm at ages 13 to 18 years were significantly associated with the presence of acute pancreatitis. Thirty-two patients, including 25 with acute pancreatitis and 7 with chronic pancreatitis, underwent follow-up measurement of pancreatic duct and serum lipase examination on at least three occasions. A good correlation between the diameter of pancreatic duct and serum lipase level was found. Thus, ultrasonography of the pancreatic duct is valuable in diagnosis and monitoring of pancreatitis in children. PMID- 11065265 TI - Sonographic features of dialysis-related amyloidosis of the shoulder. AB - This study evaluated the diagnostic role of ultrasonography in dialysis-related amyloidosis in shoulders of chronically hemodialyzed patients. Fourteen shoulders of 12 long-term hemodialysis patients were examined. All patients had been on dialysis for at least 10 years. All patients had varying degrees of pain and limitations of movement in the studied shoulders. Dialysis-related amyloidosis was the presumed diagnosis in all patients. Any patient with a history of any disease, other than dialysis-related amyloidosis, capable of producing a pathologic shoulder condition was excluded. The following parameters were studied: supraspinatus and biceps tendon thickness, tendon tears, synovial thickening, and the presence of hypoechoic material around tendons and within bursae. All shoulders had a nonhomogeneous thickening, greater than 7 mm, of the supraspinatus tendon. Seven shoulders (50%) had abnormal thickening of the biceps tendon (4 mm or greater), and two shoulders had abnormal thickening of the subscapularis tendon. Hypoechoic deposits were seen in the subdeltoid bursae and biceps sheaths in five and six shoulders, respectively. Three shoulders showed partial tears of the supraspinatus tendon, one shoulder showed a tear in the biceps tendon, and one shoulder had a tear in the subscapularis tendon. Ultrasonography is an excellent imaging modality in diagnosing the presence of dialysis-related amyloidosis in symptomatic shoulders of long-term hemodialysis patients, without having to resort to invasive procedures. The results of previous studies have been confirmed and new ultrasonographic findings described. Of particular interest is the involvement of the subscapularis tendon in dialysis related amyloidosis. Repeat ultrasonography can become an important way to follow up progression of shoulder dialysis-related amyloidosis in hemodialyzed patients. PMID- 11065266 TI - Automatic prostate boundary recognition in sonographic images using feature model and genetic algorithm. AB - This paper describes the development of a model-based boundary recognition system for transrectal prostate ultrasonographic images. It consists of two techniques: boundary modeling and boundary searching with model constraints. To achieve higher specificity of the model, a method called feature modeling is derived from the existing point distribution modeling method. To improve the robustness of the searching technique, the genetic algorithm is used. Incremental genetic algorithm with crowding replacement and binary string chromosome type was found experimentally to give good search results. It was shown that the system could recognize the boundary with considerable accuracy and consistency within a few minutes in transrectal ultrasonographic images taken from approximate middle position of the prostate. PMID- 11065267 TI - Contrast-enhanced sonography in the examination of benign and malignant adnexal masses. AB - Our objective was to characterize the properties of an intravascular ultrasonographic contrast agent in examination of adnexal masses and to compare contrast agent properties between benign and malignant adnexal tumors. Fifty eight consecutively examined women with suspected ovarian tumors were examined preoperatively by power Doppler ultrasonography, first without and then with contrast agent enhancement (Levovist). Fourteen women had ovarian cancer, 3 had borderline ovarian tumors, 18 had benign ovarian neoplasms, and 23 had functional adnexal cystic masses or endometriomas. The effect of the contrast agent was evaluated visually and by using computerized power Doppler signal intensity measurements. In visual evaluation, the brightness of the power Doppler signal and the amount of recognizable vascular areas increased in each tumor after contrast agent administration. The number of vessels in power Doppler ultrasonograms, both before and after contrast agent enhancement, was significantly higher in malignant than in benign adnexal masses, as also was the increase in the number of recognizable vessels after contrast agent administration. Contrast agent uptake time was significantly shorter in malignant than in benign tumors. No significant differences were found in the power Doppler signal intensities or their changes between benign and malignant tumors. In conclusion, use of sonographic contrast agent facilitates imaging of tumor vessels. For differentiation of benign and malignant tumors, the kinetic properties of the contrast agent, such as uptake and washout times, may have more potential than the use of the contrast agent in anatomic imaging of the tumor vessels. PMID- 11065268 TI - Echo-enhanced color Doppler sonography in children and adolescents. AB - Fifty-one patients, with a range of underlying pathologic conditions, were studied prospectively to assess the diagnostic value of echo-enhanced color Doppler sonography in the pediatric and adolescent population Their diagnoses included various tumors, vascular disorders, cerebral bleeding, pathologic conditions of small parts, and focal lesions of parenchymal organs. All patients underwent color Doppler sonography before proceeding to echo-enhanced color Doppler sonography. Diagnoses were confirmed by additional imaging (computed tomography, magnetic resonance imaging, angiography, and scintigraphy) performed as appropriate, with or without histologic study. An additional 20 children did not proceed to echoenhanced color Doppler sonography as color Doppler sonography alone was found to be sufficiently diagnostic. Levovist (SHU 508A), a contrast agent based on galactose-encapsulated air microbubbles, is approved for pediatric applications in Austria and was used as the echo-enhancing agent. Echo-enhanced color Doppler sonography was performed a total of 63 times in 51 patients (mean age, 9.8 years). Compared to color Doppler sonography, echo-enhanced color Doppler sonography either detected or enhanced visualization of pathologic conditions in 55 investigations (87.3%), yielding an overall accuracy of 95.2% (sensitivity, 95%), versus 65.7% with color Doppler sonography. One spinal arteriovenous malformation, one cerebral cavernoma, and one liver lesion were missed. The contrast material was easy to administer; no adverse reactions were observed. We conclude that echoenhanced color Doppler sonography is beneficial in pediatric sonography. It enhances visualization of vessels and perfusion, thus offering a nonionizing imaging tool for detection and follow-up evaluation of pathologic conditions with disturbed vasculature in specific cases. In infants and in persons with superficial lesions it did not offer significant advantages over color Doppler sonography. PMID- 11065269 TI - Colonic polyp in a urinary diversion causing hematuria: diagnosis on ultrasonography. PMID- 11065270 TI - Prenatal diagnosis of lipomyelomeningocele. PMID- 11065271 TI - A unifying hypothesis of Alzheimer's disease. IV. Causation and sequence of events. AB - Contrary to common concepts, the brain in Alzheimer's disease (AD) does not follow a suicide but a rescue program. Widely shared features of metabolism in starvation, hibernation and various conditions of energy deprivation, e.g. ischemia, allow the definition of a deprivation syndrome which is a phylogenetically conserved adaptive response to energetic stress. It is characterized by hypometabolism, oxidative stress and adjustments of the glucose fatty acid cycle. Cumulative evidence suggests that the brain in aging and AD actively adapts to the progressive fuel deprivation. The counterregulatory mechanisms aim to preserve glucose for anabolic needs and promote the oxidative utilization of ketone bodies. The agent mediating the metabolic switch is soluble Abeta which inhibits glucose utilization and stimulates ketone body utilization at various levels. These processes, which are initiated during normal aging, include inhibition of pro-glycolytic neurohormones, cholinergic transmission, and pyruvate dehydrogenase, the key transmitter and effector systems regulating glucose metabolism. Hormonal and effector systems which promote ketone body utilization, such as glucocorticosteroid and galanin activity, GABAergic transmission, nitric oxide, lipid transport, Ca2+ elevation, and ketone body metabolizing enzymes, are enhanced. A multitude of risk factors feed into this pathophysiological cascade at a variety of levels. Taking into account its pleiotropic regulatory actions in the deprivation response, a new name for Abeta is suggested: deprivin. On the other hand, cumulative evidence, taken together compelling, suggests that senile plaques are the dump rather than the driving force of AD. Moreover, the neurotoxic action of fibrillar Abeta is a likely in vitro artifact but does not contribute significantly to the in vivo pathophysiological events. This archaic program, conserved from bacteria to man, aims to ensure the survival of a deprived organism and controls such divergent processes as sporulation, hibernation, aging and aging-related diseases. In contrast to the immature brain, ketone body utilization of the aged brain is no longer sufficient to meet the energetic demands and is later supplemented by lactate, thus recapitulating in reverse order the sequential fuel utilization of the immature brain. The transduction pathways which operate to switch metabolism also convey the programming and balancing of the de-/redifferentiation/apoptosis cell cycle decisions. This encompasses the reiteration of developmental processes such as transcription factor activation, tau hyperphosphorylation, and establishment of growth factor independence by means of Ca2+ set point shift. Thus, the increasing energetic insufficiency results in the progressive centralization of metabolic activity to the neuronal soma, leading to pruning of the axonal/dendritic trees, loss of neuronal polarity, downregulation of neuronal plasticity and, eventually, depending on the Ca2+ -energy-redox homeostasis, degeneration of vulnerable neurons. Finally, it is outlined that genetic (e.g. Down's syndrome, APP and presenilin mutations and apoE4) and environmental risk factors represent progeroid factors which accelerate the aging process and precipitate the manifestation of AD as a progeroid systemic disease. Aging and AD are related to each other by threshold phenomena, corresponding to stage 2, the stage of resistance, and stage 3, exhaustion, of a metabolic stress response. PMID- 11065272 TI - Purification, chemical, and immunochemical properties of a new lectin from Mimosoideae (Parkia discolor). AB - A glucose/mannose-binding lectin was isolated from seeds of Parkia discolor (Mimosoideae) using affinity chromatography on Sephadex G-100 gel. The protein presented a unique component in SDS-PAGE corresponding to a molecular mass of 58,000 Da, which is very similar to that of a closely related lectin from Parkia platycephala. Among the simple sugars tested, mannose was the best inhibitor, but biantennary glycans, containing the trimannoside core, present in N glycoproteins, also seem to be powerful inhibitors of the haemagglutinating activity induced by the purified lectin. The protein was characterised by high content of glycine and proline and absence of cysteine. Rabbit antibodies, anti P. platycephala seed lectin, recognised the P. discolor lectin. However, no cross reaction was observed when a set of other legume lectins from sub-family Papilionoideae and others from families Moraceae and Euphorbiaceae were assayed with the Parkia lectins. This suggests that Parkia lectins comprise a new group of legume lectins exhibiting distinct characteristics. PMID- 11065273 TI - Chemo-enzymatic synthesis of new non ionic surfactants from unprotected carbohydrates. AB - The synthesis of new non ionic surfactants is reported. They were prepared from unprotected carbohydrates, amino acids, and fatty alcohols. These modules were linked by enzymatic esterification and transesterification reactions catalysed by lipases and proteases in organic media. PMID- 11065274 TI - Synthesis of phosphonic analogues of 4-hydroxyproline and 5-hydroxypipecolic acid. AB - Interest in non-natural amino acids is growing because of their potential biological activity. We describe in this paper, a synthesis of phosphonic analogues of 4-hydroxy proline and 5-hydroxy pipecolic acid. PMID- 11065275 TI - Expression and purification of recombinant human annexin V in Escherichia coli. AB - Human annexin V cDNA was cloned into plasmid pET19b and fused to a ten consecutive histidine tag at N-terminal. When expressed in E. coli BL21(DE3) LysS, the recombinant His10-annexin V accumulated in soluble form in the cytoplasm. By two-step chromatography, i.e., metal chelate affinity chromatography and anion exchange chromatography, recombinant His10-annexin V was purified to homogeneity on silver-stained SDS-PAGE gel. Recombinant annexin V, 7.4 mg, was obtained from a 1 litre flask culture. PMID- 11065276 TI - Growth on indium-tin oxide-coated glass enhances 32P-phosphate uptake and protein labelling of adherent cells. AB - A method to improve the efficiency of labelling of adherent cells with radioactive 32p is described. Cells are grown on a glass surface which is coated with indium-tin oxide, a commercially available, transparent material which permits excellent cell adhesion and growth. The results show that a 2 to 3-fold increase in 32p uptake by the cells can be achieved by growing cells on this material, compared to conventional tissue culture plastic. PMID- 11065277 TI - Biochemical characterization of cyanobacterial extracellular polymers (EPS) from modern marine stromatolites (Bahamas). AB - A range of biochemical characterizations were used to examine the extracellular polymeric secretions (EPS) of two cyanobacteria, Synechocystis sp. and Oscillatoria sp., isolated from marine stromatolites, Bahamas. Two-dimensional gel electrophoresis was successfully used to fractionate proteins in EPS. The results suggest that cyanobacterial EPS is composed of a network of macromolecules having different biochemical properties, which may contribute to extracellular functions. PMID- 11065278 TI - Towards a quantitative free flow electrophoresis and its application to particle size separations. AB - The possibility to quantify free-flow electrophoresis (FFE) data was explored in application to 6 negatively charged polystyrene size standards in the size range of 73 to 762 nm diameter. Peak fraction numbers in FFE were shown to be proportional to mobilities of the particles, determined by capillary zone electrophoresis in the identical buffer. Standard deviations of peak fraction numbers demonstrate a high degree of intra-experimental reproducibility while inter-experimentally, a variability of 1 to 5 peak fraction numbers within 28 fractions was found. A relative mobility (Rf) scale for peak identification in FFE based on the free mobility of the dye, SPADNS, allowed for the utilization of the entire electrophoretic migration path but failed to improve the precision of fraction numbers in view of the substantial zone spreading of the dye. Mobility differences between particles increased upon lowering the ionic strength of the electrophoretic buffer. Peak width increased with particle size in inverse relation to ionic strength. PMID- 11065279 TI - Purification and characterization of the Pasteurella haemolytica 35 kilodalton periplasmic iron-regulated protein. AB - Pasteurella haemolytica serovar A1 is the causative agent of acute fibrinohemorrhagic pneumonia also known as shipping fever. Many pathogens, including P. haemolytica, survive in their respective hosts through the up regulation of an iron acquisition system. In this study we identified, purified and characterized a 35-kDa periplasmic iron-regulated protein. The N-terminal sequence of the iron-regulated protein ANEVNVYSYRQP YLIEPMLK was identical to the deduced amino acid sequence of the ferric binding protein, FbpA, of P. haemolytica. Growth of P. haemolytica in a synthetic medium (RPMI-1640), without iron and supplemented with 50 gM 2,2' dipyridyl, facilitated the expression, isolation and purification of the native P. haemolytica FbpA. The protein was purified to homogeneity by using ammonium sulfate precipitation followed by CM Sepharose ion exchange chromatography. SDS-PAGE showed a single band with a molecular weight of 35,369. Isoelectric focusing showed multiple bands with pIs of 5.5, 5.6, 5.8, and one major band with pI of 6.4. The molecular weight obtained by electrospray mass spectrometry was 35,822. Equilibrium velocity ultracentrifugation established that the protein existed as a monomer under native conditions with an apparent molecular weight of 33,795. Analysis of secondary structure of FbpA by circular dichroism showed 42.1% alpha helical structure. This protein is the second periplasmic iron-regulated protein described for P. haemolytica. PMID- 11065280 TI - Mechanisms of beta-amyloid neurotoxicity: perspectives of pharmacotherapy. AB - One of the characteristic neuropathological hallmarks of Alzheimer's disease (AD) is the extracellular accumulation of beta-amyloid peptides (Abeta) in neuritic plaques. Experimental data indicate that different molecular forms of Abeta affect a wide array of neuronal and glial functions and thereby may lead to neuronal death in the nervous system. Whereas the fatal outcome of Abeta overproduction in transgenic cell lines, and of exogenous Abeta administration in numerous neurotoxicity models, is well established, particular facets of a complex molecular cascade by which Abeta attack neural cells are still elusive. In the present review we summarize recent knowledge on mechanisms of Abeta aggregation, its role in Abeta neurotoxicity, and binding of Abeta peptides to putative neuronal and glial receptors. Additionally, an integrative view on the interactions of Ca2+ -mediated excitotoxicity and free radical-induced oxidative stress in Abeta toxicity is provided. Furthermore, we survey advances of pharmacological investigations attempting to prevent and antagonize Abeta toxicity, or to promote neuronal regeneration following Abeta-induced neurotoxic insults. We distinguish two major classes of therapeutic approaches: conventional pharmacotherapy that employs blockade of known receptors, signal transduction pathways, and re-uptake of neurotransmitters, and direct targeting of neurotoxic Abeta by means of beta-sheet breakers, functional anti-Abeta peptides, and antibodies. Although a clinically relevant neuroprotective strategy is not yet available, sequential combination of drug regimens may provide prospects for effective antagonism of late-life Abeta burden and subsequent development of dementia. PMID- 11065281 TI - Long-term biobehavioral effects of maternal separation in the rat: consistent or confusing? AB - Over the last decades of research there has been increasing interest in endocrine and behavioral effects of postnatal environmental manipulations. A manipulation procedure that has been widely used to date is that of maternal separation. Many studies have demonstrated that, in the rat, a single or repeated separation of the pups from the mother leads to acute as well as long-term effects on endocrinology and behavior. However, reviewing the literature shows that contrary findings for almost all parameters investigated can be found. A possible explanation for this inconsistency may be the fact that maternal separation has become a collective term for a variety of extremely different experimental manipulations. Therefore, this review aims at evaluating typical effects of maternal separation in the laboratory rat by categorizing different experimental procedures. We concentrate in particular on longterm behavioral effects, although a brief summary of neuroendocrine effects is also provided. In addition, important methodological issues of maternal separation studies are discussed as a possible source for inconsistent findings. PMID- 11065282 TI - Phytobusiness requires social chemistry. PMID- 11065283 TI - 28-Norcastasterone is biosynthesized from castasterone. AB - Metabolic experiments with deuterium-labeled castasterone in seedlings of Arabidopsis thaliana, Oryza saliva and Lycopersicon esculentum, and cultured cells of Catharanthus roseus were performed, and the metabolites were analyzed by GC-MS. In all the plant species examined, [2H3]28-norcastasterone was identified as a metabolite of [26,28-2H6]castasterone, indicating that castasterone is the biosynthetic origin of 28-norcastasterone. Moreover, the natural occurrence of 28 norcastasterone and 28-nortyphasterol in seedlings of A. thaliana has been demonstrated. This is the first report of the natural occurrence of 28 nortyphasterol in plants. PMID- 11065284 TI - Tissue-specific developmental changes in cell-wall ferulate and dehydrodiferulates in sugar beet. AB - Sugar beet (Beta valgaris L.) seedlings were grown for 8-14 weeks, and then separated into leaf, petiole, inner and outer storage root and absorptive root fractions. Cell-wall ferulate and dehydrodiferulate esters were analysed by HPLC. In leaves, ferulate dimers were mostly 8-8 linked, while 8-O-4 and sometimes 8-5 linkages were most abundant in all other tissues. The total dimer content and percentage of dimerisation were much higher in the absorptive root than in other tissues. These results indicated varying patterns of ferulate and dehydrodiferulate ester content in different tissues, suggesting corresponding variations in the biosynthetic processes. When [14C]-cinnamate was applied to the leaves at 4 weeks, and [14C]-dimers measured in root cell walls at 8 and 14 weeks, a much higher proportion of 8-5 linkages was found in the [14C]-dimers than in total (non-radioactive) dimers in all parts of the root, especially at 14 weeks, indicating further complexity in the metabolism of cell-wall phenolics. PMID- 11065285 TI - A comparison of the composition of epicuticular wax from red raspberry (Rubus idaeus L.) and hawthorn (Crataegus monogyna Jacq.) flowers. AB - Epicuticular waxes have been characterised from the flowers of raspberry and hawthorn, on both of which adult raspberry beetles (Byturus tomentosus) can feed. The flower wax from both species had similar alkane profiles and also contained long-chain alcohols, aldehydes and fatty acids. The range of the carbon numbers detected for these classes of compounds was broadly similar in both but the relative amounts of each differed between species. Raspberry flower wax also contained fatty acid methyl esters, a group of compounds that has rarely been detected in plant epicuticular waxes, however, these were not observed in hawthorn flower wax. Long-chain alcohol-fatty acid esters with carbon numbers ranging from C36 to C48 were also detected in both plant species. However, an examination of their constituent acids indicated that in hawthorn the esters based on the C16 fatty acid predominated, whilst in raspberry flower wax, esters based on the C20 fatty acid were most abundant. Both species also contained pentacyclic triterpenoids, which accounted for, on average, over 16 and 48% of the total wax extracted from raspberry and hawthorn flowers respectively. In the former, ursolic and oleanolic acids accounted for over 90% of the pentacyclic triterpenes, whilst hawthorn flower wax, in addition to containing these acids, also contained high relative concentrations of both free and esterified alpha- and beta-amyrins. PMID- 11065286 TI - 6'-O-Coumaroylaloesin from Aloe castanea--a taxonomic marker for Aloe section Anguialoe. AB - The structure of 6'-O-coumaroylaloesin [2-acetonyl-8-(6-O-coumaroyl-beta-D glucopyranosyl)-7-hydrox y-5-methylchromone], a mono-ester chromone derivative in which only the 6-position of the glucosyl moiety is esterified, was determined by spectroscopic methods. The compound is a unique chemotaxonomic character restricted to the six species in Aloe section Anguialoe. PMID- 11065287 TI - Synthesis and bioactivity of 6alpha- and 6beta-hydroxy analogues of castasterone. AB - The reduction of castasterone with sodium in ethanol produced chiefly the known 6alpha-hydroxy stereoisomer, whereas reduction with sodium orohydride in methanol afforded mainly the novel 6beta-epimer. Both compounds showed variable bioactivity through four separate assays via the rice leaf lamina inclination bioassay. However, when treated with an appropriate statistical program to remove outliers, the averaged results clearly indicated that the two 6-hydroxy epimers possess comparable and significant bioactivity, which is, however, lower than that of castasterone or brassinolide. When applied together with 1000 ng of the auxin, indole-3-acetic acid (IAA), the activity of both the 6alpha and 6beta hydroxy epimers was enhanced by ca. one order of magnitude across a wide range of doses. PMID- 11065288 TI - S-alk(en)yl-L-cysteine sulfoxides, alliinase and aroma in Leucocoryne. AB - Levels of S-alk(en)yl-L-cysteine sulfoxides, alliinase and enzymatically generated pyruvic acid were determined in the bulb, leaf and scape of five species and a natural hybrid of Leucocoryne (Liliaceae), a genus of ornamental geophytes indigenous to Chile. (+)-S-Methyl-L-cysteine sulfoxide (MCSO) was present in all plant parts of all species at levels between 0.09 and 1.41 mg g( 1) fr. wt. Trans-(+)-S-(1-propenyl)-L-cysteine sulfoxide (PRENCSO) was present in plant parts of three species only (L. angustipetala, L. oadorata and L. purpurea) at levels between 0.12 and 1.82 mg g(-1) fr. wt. No other S-alk(en)yl-L-cysteine sulfoxides were detected. Alliinase (EC 4.4.1.4) was detected in the leaf, bulb and scape of L. angustipetala and L. purpurea, only in the leaves of L. coquimbensis and L. purpurea x L. coquimbensis, and only in the bulb of L. odorata. Enzymatically generated pyruvic acid was detected in all plant parts of all species at levels between trace amounts and 5.33 micromol g(-1) fr. wt. As PRENCSO is produced only in Leucocoryne species exhibiting a strong and unpleasant onion-like aroma, it is probable that the enzymatic degradation of PRENCSO is the main cause of that aroma. Consequently, Leucocoryne cultivars should be selected in species and hybrids that lack the ability to synthesise PRENCSO. PMID- 11065289 TI - Allelochemicals of the tropical weed Sphenoclea zeylanica. AB - Nine plant growth inhibitors were isolated from the tropical weed Sphenoclea zeylanica, which shows allelopathic properties. Those compounds hitherto not reported from any plant source were the isomers of cyclic thiosulfinate, (1S,3R,4R)-(+)- and (1R,3R,4R)-(+)-4-hydroxy-3-hydroxymethyl-1,2-dithiolane-1 oxides, and (2R,3R,4R)-(-)- and (2S,3R,4R)-(+)-4-hydroxy-3-hydroxymethyl-1,2 dithiolane-2-oxides. These were named zeylanoxide A, epi-zeylanoxide A, zeylanoxide B and epi-zeylanoxide B, respectively. The absolute configurations at C-3 and C-4 were elucidated by chemical synthesis of both enantiomers from L- and D-glucose. Two of the inhibitors were secologanic acid and secologanoside. and three other inhibitors were by known secoiridoid glucosides formed as artifacts during extraction with methanol. The cyclic thiosulfinates and secoiridoid glucosides completely inhibit the root growth of rice seedlings at 3.0 mM. While the specific activity of the inhibitors was not high, since they accumulated to circa 0.61% S. zelanica by dry weight, this suggests that the inhibitors are nervertheless potent allelochemicals in this weed. PMID- 11065290 TI - The role of germacrene D as a precursor in sesquiterpene biosynthesis: investigations of acid catalyzed, photochemically and thermally induced rearrangements. AB - Germacrene D is considered as a precursor of many sesquiterpene hydrocarbons. We have investigated the acid catalyzed as well as the photochemically and thermally induced rearrangement processes of germacrene D isolated from several Solidago species, which contain both enantiomers of germacrene D. Enantiomeric mixtures of sesquiterpenes of the cadinane, eudesmane (selinane), oppositane, axane, isodaucane, and bourbonane group as well as isogermacrene D were identified as main products and made available as reference compounds for structure investigations and stereochemical assignments of plant constituents. Delta amorphene, one of the rearrangement products, was identified as a natural product for the first time. The absolute configuration of gamma-amorphene was revised by correlation with the absolute configuration of germacrene D. The mechanisms of the rearrangement reactions are discussed. PMID- 11065291 TI - Long-chain alkanediols from Myricaria germanica leaf cuticular waxes. AB - In the leaf cuticular waxes of Myricaria germanica L. four different series of alkanediols were identified: (1) hentriacontanediol isomers with one functional group in the 12-position and a second group in positions ranging from 2 to 18, (2) C30-C34 alkanediols carrying one hydroxyl function on a primary and one on a secondary carbon atom. (3) homologous series of C25-C43 beta-diols predominantly with 8,10- and 10,12-functionalities, and (4) homologous series of C39-C43 gamma diols with a predominance of 8,11- and 10,13-isomers. Primary/secondary diols and gamma-diols constituted only trace portions of the total wax mixture. The hentriacontanediols and the beta-diols amounted to 3.5 and 0.6 microg per cm2 of leaf surface area, corresponding to 9 and 2% of the wax mixture, respectively. Based on the different homolog and isomer patterns of respective diol fractions, two independent biosynthetic routes leading to the hentriacontanediols and the beta-diols are proposed. PMID- 11065292 TI - Purification and characterization of deacetylipecoside synthase from Alangium lamarckii Thw. AB - Deacetylipecoside synthase (DIS), the enzyme catalyzing the condensation of dopamine and secologanin to form the (R)-epimer of deacetylipecoside, has been purified 570-fold from the leaves of Alangium lamarckii and partially characterized. The isolated enzyme is a single polypeptide with Mr 30,000, and has a pH optimum at 7.5 and a temperature optimum at 45 degrees C. The apparent Km values for dopamine and secologanin are 0.7 and 0.9 mM, respectively. DIS exhibits high substrate specificity toward dopamine, whereas neither tyramine nor tryptamine are utilized. The enzyme activity is not inhibited by its substrate dopamine, but is inhibited by alangimakine and dehydroalangimakine with similar I50 values of 10 microM. DIS presumably provides (R)-deacetylipecoside for the formation of tetrahydroisoquinoline monoterpene glucosides that also possess an (R)-configuration at the same chiral center. PMID- 11065293 TI - Lanostanes and friedolanostanes from the pericarp of Garcinia hombroniana. AB - The CH2Cl2 extract from the pericarp of Garcinia hombroniana yielded three 17,14 friedolanostanes [(24E)-3alpha-hydroxy-17,14-friedolanostan-8, 14,24-trien-26-oic acid, methyl (24E)-3alpha,23-dihydroxy-17,14-friedolanostan-8,14,24 -trien-26 oate and methyl (24E)-3alpha,9,23-trihydroxy-17,14-friedolanostan-14,2 4-dien-26 oate] and two lanostanes [3beta- and 3alpha-hydroxy-23-oxo-9,16-lanostadien-26 oic acid]. The structure of (14E)-3alpha-hydroxy-17,14-friedolanostan-8,14,24 trie n-26-oic acid was determined using spectroscopic and X-ray analyses, while the structures of the other compounds were elucidated solely from analysis of spectroscopic data. PMID- 11065294 TI - Interword spacing in Chinese text layout. AB - Three experiments using Chinese text were conducted to investigate word spacing and its effect on reading performance. In Exp. 1, a sonogram detector was used to analyze interword and intercharacter (within a word) time intervals from text read aloud by professional TV broadcasters versus college graduates. The results showed interword intervals were significantly longer than intercharacter intervals, indicating that interword spacing has psychological reality in speech. Exp. 2 examined the effect on reading performance due to separating the characters that compose a word. Separating the characters of a word did not decrease reading accuracy but did result in significantly longer reading times. Exp. 3 explored the effect of word spacing in Chinese sentences on reading performance. Analysis showed that word spacing did not affect reading accuracy, but half character and whole-character spacing significantly reduced reading time. The results of the present study suggest that word spacing in Chinese text layout enhances reading performance. Word spacing may help the reader to segment more quickly a string of characters into words and reduce the likelihood of misinterpretation. Also, ambiguity of sentence structure severely degraded reading accuracy. The implications of the results for word spacing design in Chinese text are discussed. PMID- 11065295 TI - Relationship between semantic paraphasias and related nonverbal factors. AB - Word-finding deficits are a common problem in aphasic patients. One hypothesis suggests that the difficulty that patients experience in naming objects or pictures is related to a disruption in the ability to access the lexicon. Another hypothesis suggests that these problems are caused by a disruption of the mental dictionary and the semantic representations contained in it. The main purpose of this study is to assess whether nonverbal factors such as the ability to distinguish between similar attributes of objects is related to word-use problems in aphasia. 14 adults with left hemisphere cerebrovascular accidents and some word use deficit were administered the first 30 items of the Boston Naming Test and the Conceptual Matching subtest of the Detroit Test of Learning Aptitude. Errors on the Conceptual Matching subtest correlated significantly with the number of semantic paraphasias. PMID- 11065296 TI - Factors affecting metamotivational reversals during motor task performance. AB - Reversal theory proposes that the individual's psychological state constantly switches between metamotivational state pairs (such as Apter's 1982 telic paratelic pair). Three factors are thought to affect reversals: frustration, contingent event, and satiation. Only a few studies have directly investigated these factors in sports contexts, and evidence is needed to assess support for these factors. In a laboratory setting, 24 participants performed a telic and a paratelic version of a dart-throwing task for 10 min. Participants were free to change from one task version to another as they wished, and reasons for any task changes were solicited. Task changes, indicative of reversals, were observed in 11 participants, and these were reported as due to satiation or frustration but not to contingent events. These findings may inform the structure of sessions on skill development but require confirmation in actual sports contexts. PMID- 11065297 TI - Changes in the semantic constraint of spreading activation of memory across three age groups. AB - 28 undergraduate, 34 sixth-grade, and 36 second-grade students studied target words embedded in interchangeable or noninterchangeable sentences, and then performed free recall tests. In an interchangeable sentence the word which was to be remembered and its associated word fitted sensibly, whereas in a noninterchangeable sentence the target word fitted sensibly but its word associate did not. Undergraduates recalled the target words in noninterchangeable sentences better than sixth or second graders for whom a difference was not observed (undergraduates > sixth graders = second graders). In interchangeable sentences undergraduates recalled more targets than sixth or second graders, and sixth graders recalled more than second graders (undergraduates > sixth graders > second graders). The results were interpreted as indicating changes across age groups in the semantic constraint of spreading activation of target words in memory. PMID- 11065298 TI - Overall self-confidence, self-confidence in mathematics, and sex-role stereotyping in relation to salivary free testosterone in university women. AB - This study investigated in 40 young university women the possible relationships between levels of testosterone and specific measures of overall self-confidence and self-confidence in approaching mathematics. Correlations of -.43 and -.49 with each measure of self-confidence, respectively, and level of testosterone were found in the portion of the sample displaying normal ovulatory function. There was no correlation (r=.02) between the measures of mathematics and overall self-confidence. 19 women did not display normal ovulatory function. The only significant result for the full sample was a significant correlation of -.73 between scores on the Bem Sex Role Inventory Masculinity and overall self confidence. The reasons for the high anovulatory percentage are not clear, nor are the associations which anovulation might have with either hormonal levels or the personality characteristics in question. PMID- 11065299 TI - Object concept and sleep regulation. AB - The association between infants' cognitive development and sleep regulation was investigated in 83 infants not at risk. It was found that 9-mo.-old infants with a more advanced object concept had significantly fewer sleep difficulties compared to infants with lower level of object permanence. PMID- 11065300 TI - Does background music in a store enhance salespersons' persuasiveness? AB - Background music has been studied as a key element of the store atmosphere in terms of its emotional effects; however, previous studies have shown also that music may have cognitive influence on consumers. How does music affect the salespersons' persuasive efforts within the store? To answer this question an experimental study was designed to assess the effects of four levels of arousing music conditions (no-low-moderate high arousing music). The level of pleasure of the musical pieces was controlled for. Music does not moderate significantly the effects of the salespersons on the intent to buy, but low and moderately arousing music (similarly low and moderately interesting musical pieces) does influence significantly the effects on the acceptance of the salesperson's arguments and the "desire to affiliate," i.e., to enter into communication. PMID- 11065301 TI - Spatial complexity in children's language. AB - The purpose of this research was to explore the properties of locative scenes which influence the sequence of the acquisition of spatial prepositions in English. Children ranging in age from about 2;8 to 5;6 were tested with a comprehension test involving a sentence-picture matching task. The comprehension test contained six kinds of spatial contrasts which were judged to vary in the geometric complexity of the scene. The order of acquisition was as follows: (1) into/out of & onto/off of, (2) in/on, (3) into/onto & out of/off of and through/over (around), (4) between X & Y/Y & Z, and (5) across/along. Complexity depends on a number of factors such as the number of referent objects and the nature of the relationship between the object to be located and the critical feature of the referent object. Prepositions which involve a more complex spatial geometry are more difficult for young children to comprehend. It was argued that the sequence of acquisition is partially determined by the course of conceptual development. PMID- 11065302 TI - Effective learning of writing and reading at preschool age with a multisensory method: a pilot study. AB - The readiness of preschool children (3 1/2 to 5 years old) for writing and reading was investigated using educational material specially designed for this purpose. 17 children participated in a three-month pilot program divided into monthly periods during which the language material was presented to them and subsequently, their behavior towards it was observed and recorded, 42.7% of the observations made it all three months (n = 408), showed occupation of the children with language material from 5 to 15 min. and another 49.0% occupation from 15 to 30 min. Children were involved with the language material mostly individually (92.2%). The materials that mostly attracted their interest were the tombolas and the four letter cards. Writing was the most popular activity (93.9%), followed by identifying (69.9%), reading (14.5%) and combining (6.3%). PMID- 11065303 TI - The Terpstra-Jonckheere test for ordered alternatives: randomized probability values. AB - An algorithm and FORTRAN program are presented for the Terpstra-Jonckheere test statistic and associated probability value based on a randomization routine. PMID- 11065304 TI - Contextual mediation of perceptions during hauntings and poltergeist-like experiences: a replication and extension. AB - This study is a replication of the experiment by Lange, Houran, Harte, and Havens (1996 on contextual variables, in which hallucinations appear to be affected by the environmental context. These contextual variables are influential in the reporting of haunting and poltergeist-like episodes. This study extended the previous study by adding new factors of time of day, climactic conditions, and emotional feelings. These were analyzed for a different sample, looking for further congruency between experiential content and the context. The sample (N=8431 were reports found on the Internet and in one book. The Lange, et al. study was replicated in that contextual variables were identified in 99.2% of the reports, the content of the reports was judged to be consistent with the nature of the contextual variables in 58.8% of the reports, and contextual variables were related to the percipients' state of arousal and the modalities of experience. PMID- 11065305 TI - Internal consistency reliability of the tactual performance test trials. AB - Internal consistency reliabilities were calculated for the Tactual Performance Test Preferred Hand (n=300), Nonpreferred H and (n=302), and Both Hands (n=314) trials, and Total Time. Reliabilities are reported for the total sample and three groups: normal, undiagnosed patients sent for assessment, and alcoholic persons. The reliability coefficients ranged from .59 to .90. PMID- 11065306 TI - Can different levels of S-R practice influence sequence learning? An investigation into the context of a perceptual-motor sequence learning task. AB - The main aim of the present study was to evaluate on a serial reaction time task the effect of stimulus-response (S-R) practice on sequence learning. The experiment used a pointing task which allowed recording reaction times and movement times. The basic manipulation consisted in varying the amount of S-R practice prior to sequence practice. Two main findings from this study may be highlighted. Firstly, the benefit from extensive S-R practice was mainly observed in the random practice phase. Secondly, S-R learning and sequence learning were reflected by different components of performance. The movement times were selectively sensitive to the acquisition of S-R regularities whereas the reaction times were selectively sensitive to the acquisition of sequence regularities. The implications of these results on the comprehension of the sequence learning mechanism were then discussed. PMID- 11065307 TI - Retention of relative force in the scaling of a serial force pattern. AB - The present study was designed to examine the retention of relative force in the scaling of a serial force pattern in finger-tapping sequences. On practice trials, 11 male college students tapped a force plate connected to strain gauges which provided feedback. On test trials, subjects recalled both the force pattern (100 g-100 g-100 g-300 g) and intertap interval (400 msec.) acquired during practice without feedback (recalled task) and then produced a halved (halved task) or doubled-force (doubled task) at the fixed intertap interval. Analysis showed that, although there was no difference for absolute forces between the recalled task and the halved task, the forces at the doubled task were three times as great as those at the recalled task. For relative forces, on the other hand, although there was no difference among the three tasks, the force ratios were closer to 1:1:1:2 than 1:1:1:3. This indicated that the scaling of force pattern was a more difficult adaptive task than that of the intertap interval in the previous study (Inui, Ishida, & Yamanishi, 1999). PMID- 11065308 TI - Middle-school students of United Arab Emirates: effects of heterogeneous small group work on attitudes toward mathematics. AB - This study compared the effects of two instructional strategies, small heterogeneous cooperative learning experience versus lecture and discussion, on students' attitudes toward mathematics. 54 boys and 57 girls in Grade 8 of four middle-school mathematics classes participated. Two classes (57 students) were taught using a cooperative learning method and the other two classes (54 students) were taught using traditional lecture and discussion. Differences between attitudes of boys and girls were also investigated and discussed in the light of Arabic culture. The results suggested that cooperative learning might be a valuable method with which to teach mathematics concepts to boys. PMID- 11065309 TI - Exploring competitive orientation in a group of athletes participating in the 1996 paralympic trials. AB - The purposes of this study were to test (1) whether athletes with congenital disabilities exhibited different competitive orientations than athletes with disabilities acquired during their lifespans and (2) whether male athletes with disabilities exhibited different competitive orientations than their female peers. 54 paraplegic, quadriplegic and amputee athletes competing in the 1996 Paralympic Track and Field Trials completed the Sport Orientation Questionnaire. No mean differences were found between men and women, athletes with different onsets of their disabilities across the lifespan, between adolescents and adults, and between athletes with different severity classifications on the Goal orientation, Competitiveness, and Desire to win scales. Larger studies are encouraged to examine competitive orientation, as well as scores on tests specifically constructed to be administered to athletes with disabilities. PMID- 11065310 TI - Judgments of grammaticality of sentences with a differing number of arguments: a comparison of English and Japanese speakers. AB - This study explored native speakers' linguistic intuition as revealed in judging the grammaticality of sentences. Speakers of English (n = 36) and Japanese (n = 57) judged the relative grammaticality of sentences involving a verb which occurred together with one, two or three arguments. Findings showed that English speakers were more affected by the number of arguments in sentences. They judged sentences having three arguments as grammatical most often and ones having one or two arguments as grammatical less often. However, Japanese speakers gave rather similar and more grammatical judgments regardless of the number of arguments in sentences. The findings indicate a difference in tightness of argument structure in the two languages even when the sentences judged are simple sentences and they are given without any sentential context. PMID- 11065311 TI - An analysis of employee voting behavior in multi-union certification elections among unaffiliated unions. AB - Unions experience higher success rates in multi-union certification elections than in single-union elections. In multi-union elections, unions have the highest victory rate when the two unions competing against each other are unaffiliated unions. The outcomes of the multi-union election process involving unaffiliated unions are examined with respect to election type and type of competition among unions. PMID- 11065312 TI - Effects of short-term psychological stress on the time and frequency domains of heart-rate variability. AB - The purpose of this study was to test the hypothesis that short-term psychological stress produces significant changes in sympathovagal activity. A simple, noninvasive method was used measuring the timing and frequency of heart rate variability (HRV). 30 normal healthy subjects were assigned into two age- and sex matched groups. In the experimental condition a 5-min. psychological stress test, predominantly based on the Stroop Word Color Conflict Test, was employed in a competitive setting and included a financial inducement to produce psychological strain. Analysis showed that during psychological stress a significant reduction in the timing and frequency of heart rate variability was observed. The standard deviation of interbeat intervals decreased. A significant increase in heart rate was also observed. Within the frequency domain, a significant reduction in the high frequency component of HRV and a significant increase in the low frequency component were observed. There was also a significant increase in the low frequency to high frequency ratio. Self evaluation of physical tension and emotional state measured by visual analog scales also showed significant increases following psychological stress. No significant differences were observed on any variables within the control group. The results indicate a shift towards sympathetic predominance as a result of parasympathetic withdrawal and demonstrates that this psychological stress test is effective in provoking a characteristic defence-arousal reaction. This simple, cost-effective method of analysing heart rate variability is suitable for detection of short-term changes in sympathovagal balance. PMID- 11065313 TI - Sex differences in identifying the facial affect of normal and mirror-reversed faces. AB - The influences of sex and lateralized visual hemispace bias in the judgment of the emotional valence of faces during a free viewing condition are evaluated. 73 subjects (aged 18 to 52 yr.) viewed videotaped facial expressions of emotion in normal and mirror-reversed orientation and classified each face as a positive, negative, or neutral expression. There was a significant interaction between the sex of the rater and the orientation of the face that influenced the proportion of correct classifications. Male and female perceivers did not differ in the accuracy of their affect judgments for faces viewed in normal orientation, whereas reversal of the orientation of the faces resulted in a significant enhancement of accuracy judgments for the males but not the females. The results suggest greater cerebral lateralization of perceptual processes in males. PMID- 11065314 TI - Academic performance and participation in physical activity by secondary school adolescents. AB - This study investigated the relationship between adolescents' academic performance and participation in physical activity. 232 boys and girls from Years 8-11 (ages 13-16 years) were randomly selected, and their academic performance was assessed on previous examination scores in English, Mathematics, and Science. Participants were also asked to list all the sports based physical activities in which they normally participated during a typical week and to indicate how many times per week they took part in each activity and the duration of each. Overall, no significant correlations were found, although weak negative correlations were recorded between the amount of time (in minutes) in sport and exercise and English scores for children ages 13, 14, and 16 years. A similar association was also noted for Science scores of children 16 years old. PMID- 11065315 TI - Sex differences and individual consistency in voice identification. AB - Results were combined from 5 experiments by Cook in 1998 with 728 participants who listened to one male voice and one female voice each saying a sentence, then attempted to recognise the voices from line-ups of six voices presented a week later. While 352 male listeners did not differ significantly in recognising female and male voices (38% correct vs 41%), 376 female listeners were significantly more likely to recognise female than male voices (51% vs 43% correct). There was no evidence for individual differences in voice recognition in that listeners who recognised the male voice were no more likely to recognise the female voice than those who failed to recognise the male voice. PMID- 11065316 TI - Effect of an audience on learning a novel motor skill. AB - The purpose of this study was to assess effects of an audience on learning a novel motor skill. Subjects (N=64) were randomly assigned to one of four experimental conditions and administered 15 30-sec. trials with 30 sec. intertrial periods on a pursuit rotor task on two different days. Comparison of Time-on Target performance between conditions indicated that the No Audience condition had significantly higher performance than the Audience condition in Session 1. Comparison of Absolute Retention and Final Retention scores among the four experimental conditions in Session 2 after 48 hr. yielded no significant differences attributable to the presence of an audience, thus supporting the hypothesis that an audience would have no effect on learning. PMID- 11065317 TI - Are better eye movements an advantage in ball games? A study of prosaccadic and antisaccadic eye movements. AB - The aim of this study was to compare prosaccadic and antisaccadic eye movements of experts in ball sports and controls. In the prosaccadic and antisaccadic task, subjects made saccades respectively towards and away from a suddenly appearing stimulus. By means of infrared-oculography, we compared horizontal eye movements of experts (n=18) and controls (n=20). Experts had shorter overall saccadic latencies, but significantly shorter latencies occurred only on the antisaccade task, not on the prosaccade task. Our findings seem to support the concept that prosaccades and antisaccades have different underlying mechanisms and that expertise in ball games mainly improves antisaccadic performance in terms of latency and variability. PMID- 11065318 TI - Performance of a psychomotor skill following rest, exercise at the plasma epinephrine threshold and maximal intensity exercise. AB - The purpose of this study was to examine the effect of exercising at the epinephrine threshold and at Maximum Power Output on the performance of a skill that requires both decision-making and motor performance. Participants (N=12) undertook an incremental test to exhaustion from which their epinephrine threshold and Maximum Power Output were calculated. They were then examined on a soccer skill test following rest and exercise that was previously determined to elicit their epi nephrine threshold and Maximum Power Output. The soccer test examined the participants' speed and accuracy of response. Speed of response was measured by voice reaction time and whole body reaction time. No significant effects of exercise were shown for any of the variables. The need for further research using more complex skill tests and the use of discontinuous exercise protocols, rather than continuous ones, is recommended. PMID- 11065319 TI - Ambition as a motivational basis of organizational and professional commitment: preliminary analysis of a proposed career advancement ambition scale. AB - This study proposes that the ambition to advance in one's career may serve as a motivational basis of organizational and professional commitment. In support of this notion, preliminary evidence of the reliability and construct validity of a proposed Career Advancement Ambition Scale is presented, and exploratory analyses of a secondary data set show that the scale and original scales of organizational and professional commitment can predict turnover intentions. Limitations and implications for future research are discussed. PMID- 11065320 TI - Geophysical variables and behavior: LXXXXIX. The influence of weather on suicide in Hong Kong. AB - Summer peaks in suicide for both sexes and ages groups (> or = 55 years) in Hong Kong were ascertained. Barometric pressure had a very weak negative relation with suicide rate. PMID- 11065321 TI - An automated administration of Corsi's Block-tapping Test. AB - Corsi's Block-tapping Test has been a clinically useful test of visuospatial memory. Participants (15 men, 15 women) completed an automated and a manual version of the Block-tapping Test, the findings for which suggest that the automated and manual versions gave very similar scores. PMID- 11065322 TI - Magnitude estimation of manually assessed elastic stiffness: stability of the exponent. AB - Magnitude estimations were obtained for manual assessments of pure elastic stiffness stimuli (metal springs). 20 subjects of varied experience in manual assessment of spinal stiffness volunteered to participate. The mean exponent of the power function relating perceived magnitude of elastic stiffness to measured physical magnitude was 1.65. Exponents varied across the 20 individuals but were stable across testing sessions held at least 2 weeks apart, and the size of the exponent was not related to prior experience. PMID- 11065323 TI - Validation of a single-item measure of usual physical activity. AB - The purpose of this study was to assess the reliability and validity of a single item measure of Usual Physical Activity and to assess its usefulness as a physical activity tool for perimenopausal women. 188 perimenopausal women participated (age: M = 47 yr., SD = 3; range = 40-55). Data were collected using the Women's Health Assessment Scale, the Physical Activity Questionnaire, the Perimenopause-related Quality of Life Scale, a health history and demographic questionnaire, and the rater. Scores were stable over a 2-wk. interval. Convergent validity was supported by a correlation of .66 between ratings on Usual Physical Activity and the Physical Activity Questionnaire. Concurrent validity was supported by the association of the rating of Usual Physical Activity with three parameters of Body Mass Index, psychosomatic symptoms, and perimenopause related quality of life, known to be associated with physical activity. Highly active women had a lower Body Mass Index than less active and inactive women. Active women tended to report fewer and less distressing psychosomatic symptoms and better quality of life. These findings support the use of rating of Usual Physical Activity to classify perimenopausal women into categories of physical activity. PMID- 11065324 TI - Copying strategies for patterns by children and adults. AB - In the present study, eve movements during the copying of a pattern were analyzed to compare visual strategies of adults and children. Subjects had to build an accurate copy of spatial block patterns. Tested variables were the incidence and the duration of ocular dwelling in the pattern area (where the pattern to be copied was located), the work area (where the copy was made), and the source area (where the blocks were that could be used for creating the copy). Furthermore, to unravel employed strategies, sequences of dwelled areas were investigated. Previous studies reported that adults employ repetitive visual scanning strategies to accomplish the task instead of strategies depending upon an internal representation. The present results show that the II children, within the ages of 7 to 12 years, made more eye movements and fixations of longer duration during copy tasks than the 11 adults. The visual strategies of the children were highly comparable to those of adults. Memory was restricted to one block, while color and location seemed to be remembered together. PMID- 11065325 TI - Correlates of rates of suicide and homicide committed by different methods. AB - The social correlates of suicide rates by each method over time in the USA from 1950 to 1985 were very different, suggesting that suicides by different methods may not be sociologically identical behaviors. Homicide rates by each method, however, showed a more consistent pattern of social correlates. PMID- 11065326 TI - Phonoemotional profiling: a description of the emotional flavour of English texts on the basis of the phonemes employed in them. AB - Research employing three large lists of words rated along emotional dimensions (total N = 15,761 words) supported a prior claim that most phonemes have a distinct emotional character. Different phonemes tended to occur more often in different types of emotional words. When phonemes were grouped along eight radii in a two dimensional emotional space defined by Pleasantness and Activation (Pleasantness, Cheeriness, Activation, Nastiness, Unpleasantness, Sadness, Passivity, and Softness), it became possible to draw profiles of texts in terms of their preferential use of different classes of phonemes. Four experiments were performed to illustrate the manner in which phonemes in nonsense words are related to emotion, and evidence of the validity of character assignments was investigated and received support in three further analyses. The emotionality of phonemes was related to both place and manner of articulation and to properties of the auditory signal itself. Phonoemotional profiles were drawn for several types of material and provided supporting evidence for the validity of the assignment of emotional character to phonemes. PMID- 11065327 TI - Mood and emotion in sport: a response to Jones, Mace, and Williams (2000). AB - Examination of the mood and performance relationship in sport has been an important line of investigation in sport psychology for over 20 years. Recent research has challenged the notion that the Profile of Mood States is the instrument of choice. It has also moved away from the notion that successful performance is associated with an 'iceberg' profile (Morgan, 1980), arguing that researchers should consider mood and emotion as distinguishable constructs (Jones, Mace, & Williams, 2000; Lane & Terry, 2000. In the present paper, I expand the discussion on mood research in sport by re-examining some of the findings of Jones, et al. in the light of recent work. PMID- 11065328 TI - Discriminating the sex of faces by 6- and 8-mo.-old infants. AB - The goal of this study was to ascertain at what age infants could discriminate male and female faces using only the internal features of the face. The habituation-dishabituation technique was used to estimate infants' discrimination between male and female faces. Analysis showed that 8-mo.-old infants discriminated female and male faces, whereas 6-mo.-old infants did not, but showed an asymmetry in discrimination. 6-mo.-old infants who were habituated to the female face fixated consistently longer on the novel male face in test trials, so sex discrimination was complete but not observed after habituation to male faces. Data are discussed in relation to the role of experience in face discrimination. PMID- 11065329 TI - Actors', partners', and observers' perceptions of sarcasm. AB - This study compared actors', partners', and observers' perceptions of the amount of sarcasm used by participants (n = 80) in videotaped conversations. Significant differences were found among perceptions of actors, partners, and observers. Of the three perspectives, actors perceived themselves as using the greatest amount of sarcasm, followed by partners' perceptions of actors. Observers perceived actors as using the least amount of sarcasm. Correlations conducted to assess whether partners and observers recognized actors' individual attempts at sarcasm during the conversations were generally low. PMID- 11065330 TI - A revised spatial serial learning and memory procedure using Corsi's Block tapping apparatus. AB - The purpose of this study was to use Corsi's Block Tapping Test as a spatial analog of Benton's Serial Digit Learning Test, using the cognitive neuroscience approach utilized in the California Verbal Learning Test. 60 normal participants, ages 19-52 years, were included and administered an 8-block sequence for 9 trials or until they recalled the entire sequence for 3 consecutive errorless trials. The score was the number of blocks tapped in the correct serial order. An interference trial was administered. Following a 10-min. delay, free recall of the original sequence, cued recall, and recognition measures were obtained. Retroactive interference was significant, but no proactive interference emerged. Scores showed a strong primacy effect. Most participants who learned the sequence to the criterion of three successive errorless trials recalled the sequence after the 10-min. delay. Scores on the cued recall and recognition trials tended to support their validity as less demanding retrieval tasks. The use of this spatial learning and memory procedure allows finer discriminations among nonverbal memory deficits and may facilitate direct comparisons with scores on verbal memory tasks such as Serial Digit Learning and the California Verbal Learning Test. PMID- 11065331 TI - Reference group data for the Reitan-Indiana Neuropsychological Test Battery for Young Children. AB - This study presents a data set for a reference group on the Reitan-Indiana Neuropsychological Test Battery for Young Children. The data set is based on a sample of 224 children, ages 5 to 8 years, referred to a special services cooperative for academic or behavioral concerns during the years 1980 through 1993. Data are presented in terms of sample size, means, standard deviations, diagnostic classifications, and population characteristics. Previously published data sets are reviewed in comparison to this newly acquired data set. Potential advantages of this data set include the larger sample, contemporary data collection, and a sample drawn from a United States school-referred population. PMID- 11065332 TI - A force/displacement analysis of muscle testing. AB - Manual muscle testing procedures are the subject of a force and displacement analysis. Equipment was fabricated, tested, and employed to gather force, displacement, and time data for the purpose of examining muscle-test parameters as used by clinicians in applied kinesiology. Simple mathematical procedures are used to process the data to find potential patterns of force and displacement which would correspond to the testing of strong and weak muscles of healthy subjects. Particular attention is paid to the leading edge of the force pulses, as most clinicians report they derive most of their assessment from the initial thrust imparted on the patient's limb. An analysis of the simple linear regression of the slope (distance vs force) of the leading edge of a force pulse indicates that a significantly large slope is indicative of weak muscles (as perceived by the clinician), and a small slope is indicative of strong muscles. Threshold criteria for slopes are specified to create a model that may discriminate between strong and weak muscles. The model is accurate 98% of the time compared to judgments of clinicians with more than 5 years of experience but is considerably lower for clinicians with less than five years of experience (64%). this accuracy rate indicates that the model is reliable in predicting the clinician's perception of muscle strength, and it also indicates that the testing procedure for muscle strength used by experienced clinicians in applied kinesiology are reliable. The experiment lays the groundwork for studies of the objectivity of muscle-strength assessment in applied kinesiology. PMID- 11065333 TI - Trait anxiety among students in a college golf class. AB - This study examined trait anxiety among students (N=80) participating in a beginning golf activity class. Analysis showed no differences in anxiety scores by sex or playing experience; however, performance differences were noted as students with moderate scores performed better on golf evaluations than did those with high anxiety scores. Research should be directed to assess students' perceptions of anxiety and its effects. PMID- 11065334 TI - Executions as a deterrent to homicide. AB - For the period 1977-1992, the number of homicides in a state tended to decrease more often after a year of no executions than after a year with one or more executions, the opposite of a deterrent effect. PMID- 11065335 TI - Pregnant women's social status, stress, self-esteem, and their infants' sex ratio at birth. AB - A reported association of social status of parents with infants' sex ratio at birth and of psychological stress (score on Beck Depression Inventory) with sex ratio were not supported by our analysis, but the possibility of an association between scores on the Rosenberg Self-esteem scale and sex ratio at birth for a sample of 385 pregnant women showed that women who have given birth to boys scored lower on self-esteem during pregnancy than those who have given birth to girls. Some explanations are reviewed to discuss this unforeseen association. PMID- 11065336 TI - Always the whole and its parts. PMID- 11065337 TI - Transgenic mice: an irreplaceable tool for the study of mammalian development and biology. AB - Stable integration into the mouse genome of exogenous genetic information, i.e., the creation of transgenic mice, has become a privileged way of analyzing gene function in normal development and pathology. Both gene addition and gene replacement may be performed. This has allowed, in particular, the creation of mice in which precise mutations are introduced into a given gene. Furthermore, in recent years, strategies that induce the expression of a mutation in a given type of cell and/or at a given time in development have been developed. Thus, the transgenic methodology affords a unique and irreplaceable tool for the study of mammalian development and biology and for the creation of animal models for human genetic diseases. PMID- 11065338 TI - Inducible gene expression and gene modification in transgenic mice. AB - Animal transgenesis has proven to be useful for physiologic as well as pathophysiologic studies. Animal models with conditional expression of a transgene of interest or with a conditional gene mutation can be generated. This permits spatial and temporal control of the expression of the transgene or of gene mutations previously introduced by gene targeting. These approaches allow the generation of models suitable for physiologic analysis or models mimicking disease states. PMID- 11065339 TI - Effect of the genetic background on the phenotype of mouse mutations. AB - An increasing number of scientific articles report that the phenotype of a given single gene mutation in mice is modulated by the genetic background of the inbred strain in which the mutation is maintained. This effect is attributable to so called modifier genes, which act in combination with the causative gene. The modulation of the phenotype can be major, as exemplified in the case of several mouse models of polycystic kidney disease. Because of the existence of inbred strains and the possibility of developing congenic strains, the effect of the genetic background can be analyzed in mice, including the identification of major modifier genes. Furthermore, by transferring a given mutation into different genetic backgrounds, mouse models can be manipulated with the aim of more accurately mimicking specific features of human diseases. PMID- 11065340 TI - Wilms' tumor suppressor gene WT1: from structure to renal pathophysiologic features. AB - Normal development of the kidney is a highly complex process that requires precise orchestration of proliferation, differentiation, and apoptosis. In the past few years, a number of genes that regulate these processes, and hence play pivotal roles in kidney development, have been identified. The Wilms' tumor suppressor gene WT1 has been shown to be one of these essential regulators of kidney development, and mutations in this gene result in the formation of tumors and developmental abnormalities such as the Denys-Drash and Frasier syndromes. A fascinating aspect of the WT1 gene is the multitude of isoforms produced from its genomic locus. In this review, our current understanding of the structural features of WT1, how they modulate the transcriptional and post-transcriptional activities of the protein, and how mutations affecting individual isoforms can lead to diseased kidneys is summarized. In addition, results from transgenic experiments, which have yielded important findings regarding the function of WT1 in vivo, are discussed. Finally, data on the unusual feature of RNA editing of WT1 transcripts are presented, and the relevance of RNA editing for the normal functioning of the WT1 protein in the kidney is discussed. PMID- 11065341 TI - Secreted molecules in metanephric induction. AB - Nearly 50 yr. ago, Clifford Grobstein made the observation that the ureteric bud induced the nephrogenic mesenchyme to undergo tubulogenesis. Since that discovery, scientists have attempted to characterize the molecular nature of the inducer. To date, no single molecule that is both necessary and sufficient for nephric induction has been identified. Because of recent insights regarding the role of several secreted molecules in tubulogenesis, it has become necessary to revise the classic model of metanephric induction. The studies of the classic ureteric inducer performed to date have most likely been characterizations of a mesenchyme-specific inducer, Wnt-4, and its role in tubulogenesis. Ureteric induction most likely involves a series of distinct events that provide proliferative, survival, and condensation signals to the mesenchyme, integrating the growth of the ureteric system with tubulogenesis. PMID- 11065342 TI - Mouse models of nitric oxide synthase deficiency. AB - Knockout mice for each of the three nitric oxide (NO) synthase (NOS) genes have been generated. Their phenotypes reflect the roles of each NOS isoform in physiologic and pathologic processes. This article reviews how neuronal NOS (nNOS) and endothelial NOS (eNOS) knockout mice have contributed to our knowledge of the roles of NO in cerebral ischemia, cardiovascular processes, and the autonomic nervous system. In some instances, the effects of NO produced by one isoform antagonize the effects of NO produced by another isoform. For example, after cerebral ischemia, the nNOS isoform is involved in tissue injury, whereas the eNOS isoform is important in maintaining blood flow. All three isoforms are expressed in the respiratory tract, but only the nNOS isoform appears to be involved in modulating airway responsiveness and only the inducible NOS isoform appears to respond to antigen stimulation. In the cardiovascular system, endothelial NO is important for vascular tone, systolic and diastolic cardiac function, vascular proliferative responses to injury, platelet aggregation, and hemostasis. PMID- 11065343 TI - Mechanisms mediating the renal profibrotic actions of vasoactive peptides in transgenic mice. AB - Transgenic mice are useful tools to investigate the mechanisms of the renal profibrotic actions of endothelin and angiotensin II. The overexpression of angiotensinogen and renin genes induces renal sclerosis independently of changes in systemic hemodynamics. The same results are observed when the endothelin-1 gene is overexpressed. Transgenic mice harboring the luciferase gene, under the control of the collagen I alpha2 chain promoter, and made hypertensive by induction of a nitric oxide (NO) deficiency have been studied. In this strain of mice, luciferase activity is an early index of renal and vascular fibrosis. Luciferase activity was increased in preglomerular arterioles and glomeruli when mice were treated with N:(omega)-nitro-L-arginine methyl ester, an inhibitor of NO synthases. Bosentan (an endothelin receptor antagonist) was as efficient as losartan (an AT1 receptor antagonist) in preventing renal fibrosis, although it did not decrease BP. In short-term experiments, angiotensin II produced an increase in luciferase activity that was entirely prevented by losartan but also by bosentan. It can be concluded that, during chronic inhibition of NO, the collagen I gene is activated, which contributes to the development of nephroangiosclerosis and glomerulosclerosis. Angiotensin II plays a major role in this fibrogenic process, and its effect is at least partly independent of systemic hemodynamics and mediated by the profibrotic action of endothelin-1. PMID- 11065344 TI - Scnn1 sodium channel gene family in genetically engineered mice. AB - The amiloride-sensitive epithelial sodium channel is the limiting step in salt absorption. In mice, this channel is composed of three subunits (alpha, beta, and gamma), which are encoded by different genes (Scnn1a, Scnn1b, and Scnn1c, respectively). The functions of these genes were recently investigated in transgenic (knockout) experiments, and the absence of any subunit led to perinatal lethality. More defined phenotypes have been obtained by introducing specific mutations or using transgenic rescue experiments. In this report, these approaches are summarized and a current gene-targeting strategy to obtain conditional inactivation of the channel is illustrated. This latter approach will be indispensable for the investigation of channel function in a wide variety of organ systems. PMID- 11065345 TI - Comparative roles of the renal apical sodium transport systems in blood pressure control. AB - Human genetic studies suggest that the genes encoding renal apical Na(+) transport proteins play an essential role in the control of extracellular fluid volume and BP. Mice with mutations in each of these genes provide the unique opportunity to directly assess their respective involvement in fluid homeostasis and BP control in vivo. Inactivation of either the epithelial Na(+) channel (ENaC) or the Na(+)-Cl(-) cotransporter decreases BP to the same extent in mice fed a low-salt diet, despite a more pronounced perturbation of fluid homeostasis in ENaC-deficient mice. In contrast, inactivation of Na(+)/H(+) exchanger 3 (NHE3) or the Na(+)-K(+)-2Cl(-) contransporter reduces BP with a normal-salt diet and renders mice unable to survive with a low-salt diet. Therefore, the general conception that ENaC in the collecting duct is the main renal controller of Na(+) balance and extracellular fluid volume should be tempered. For example, NHE3 in the proximal convoluted tubule seems to play a more substantial role in the control of fluid homeostasis. The overall effect of NHE3 inactivation on BP may also involve absorptive defects in the intestine and colon, where the exchanger normally reabsorbs significant amounts of Na(+) and water. PMID- 11065346 TI - Hepatocyte nuclear factor 1, a transcription factor at the crossroads of glucose homeostasis. AB - Hepatocyte nuclear factor 1 (HNF1) is a transcription factor involved in the regulation of a large set of hepatic genes, including albumin, beta-fibrinogen, and alpha1-antitrypsin. HNF1 is expressed in the liver, digestive tract, pancreas, and kidney. Mice lacking HNF1 exhibit hepatic, pancreatic, and renal dysfunctions. HNF1-deficient mice fail to express the hepatic phenylalanine hydroxylase gene, giving rise to hyperphenylalaninemia. Renal proximal tubular reabsorption of glucose, phosphate, arginine, and other metabolites is affected, producing severe renal glucosuria, phosphaturia, and amino aciduria. Homozygous mutant mice also exhibit a dramatic insulin secretion defect. This dysfunction resembles that exhibited by patients with maturity-onset diabetes mellitus of the young type 3, who carry mutations in the human HNF1 gene in the heterozygous state. These data show that HNF1 is a major regulator of glucose homeostasis, regulating the expression of genes that are expressed in the liver, kidney, and pancreas. PMID- 11065347 TI - Using transgenic mice to analyze the mechanisms of progression of chronic renal failure. AB - An understanding of the mechanisms underlying the formation of renal lesions is necessary for the development of strategies aiming to delay the progression of chronic renal failure. The generation of transgenic mice in the past 20 years has contributed significantly to the study of this phenomenon. Overexpression and/or inactivation of single factors in renal tissue demonstrated that molecules such as growth factors, proto-oncogenes, and renin-angiotensin system elements play major roles in renal deterioration. Several mouse models of renal injury have been developed in the past 10 yr. Transgenic mice that exhibit a normal phenotype under physiologic conditions allow analysis of the roles of single factors in the progression of chronic renal failure when renal injury models are used. Using this strategy, it was demonstrated that vascular adaptation, which is a process that involves the endothelin/nitric oxide balance, is essential for the survival of mice after nephron reduction and that the epidermal growth factor/activator protein-1/Bcl-2 pathway is involved in the development of renal lesions after renal injury, possibly via adjustment of the proliferation/apoptosis balance. Moreover, it was demonstrated that selective inhibition of epidermal growth factor signaling in the kidney successfully prevents the progression of chronic renal failure. These results indicate the power of transgenesis for elucidation of the pathogenesis of renal disease. PMID- 11065348 TI - Critical aspects of viral vectors for gene transfer into the kidney. AB - Viral vectors have been used in vitro and in vivo for more than a decade, with some significant results in specific situations, e.g., when recombinant adeno associated virus is used for the long-term transduction of skeletal muscle in coagulation factor IX-deficient patients. However, the kidney has been quite difficult to transduce with any viral vector currently available. When viral transduction occurs, it is often heterogeneous, transient, and eventually associated with immune and toxic side effects. However, recombinant adeno associated virus and lentiviral vectors remain to be fully evaluated in the kidney; the former is small enough to be filtered through the glomerular basement membrane. This may be critical, because glomerular filtration is required for DNA complex-mediated transduction of tubular cells. An alternative to in situ renal gene transfer is secretion of a therapeutic protein from a distant site, such as skeletal muscle. Several examples provide evidence that this could be a clinically relevant approach. It also may allow accurate determination of the pathophysiologic mechanisms involved in the establishment and maintenance of experimental glomerulonephritis. PMID- 11065349 TI - Renal transfer of genetically engineered cells. AB - For many years, ex vivo gene transfer has been used for genetic manipulation of various organs. In the kidney, ex vivo gene transfer was reported using mesangial cells and macrophages. In rats, cultured cells injected into the renal artery are accumulated selectively in the glomerulus. With this approach, it is possible to transfer genetically engineered cells to normal and diseased glomeruli. The transfer of genetically engineered cells to glomeruli can be used for several purposes. With the use of resident glomerular cells engineered in vitro, it is possible to examine how the cells that overexpress certain genes behave differently in normal and diseased glomeruli. Both gain-of-function and loss-of function strategies are useful for this purpose. For the latter, stable expression of antisense cDNA, ribosomes, or dominant-negative mutants is available. By transfer of engineered cells producing secretory, recombinant proteins, it is possible to modify glomerular microenvironment in vivo. Transfer of genes encoding therapeutically relevant molecules could be useful for therapeutic intervention. Transfer of engineered leukocytes to the glomerulus also allows investigation of cross talk between leukocytes and resident cells. Transfer of stimulated leukocytes is useful for investigation of the pathologic actions of infiltrating cells on glomerular structure and function. Leukocytes in which certain gene functions are selectively reinforced or deleted would be useful for elucidation of the exact functions of leukocyte-associated genes in glomerular diseases. This article summarizes current experience with the adoptive transfer of engineered cells to the glomerulus for investigation of and therapy for glomerular diseases. PMID- 11065350 TI - Delivering erythropoietin through genetically engineered cells. AB - Erythropoietin (Epo) is a glycoprotein hormone produced by genetic engineering. Many pathologic conditions could benefit from its administration, such as chronic renal failure or hemoglobinopathies. Epo secretion from genetically modified tissued could be proposed to patients only if the protocol is low cost and low risk. For that purpose, retroviral vectors and adeno-associated vectors expressing the Epo cDNA were developed. Gene transfer was performed into skeletal muscles. To avoid polycythemia, a tetracycline-regulated system was used to control the levels of protein secretion in vivo. beta-thalassemias are among diseases that could benefit from an Epo gene transfer. beta-thalassemias are attributable to deficient synthesis of beta-globin and accumulation of unpaired alpha-chains. Stimulation of fetal globin synthesis is one strategy to correct the globin chain imbalance. There is evidence that Epo could play this role. In a mouse model of beta-thalassemia, an adeno-associated vector expressing the Epo cDNA was injected intramuscularly. Epo was secreted continuously during at least 1 yr. Erythropoiesis was improved in those mice by increasing the synthesis of fetal hemoglobin. PMID- 11065351 TI - Closing in on Chlamydia and its intracellular bag of tricks. PMID- 11065352 TI - Virulence and drug susceptibility of Mycobacterium celatum. AB - The virulence and drug susceptibility of a clinical isolate of Mycobacterium celatum which showed smooth transparent (ST) and smooth opaque (SO) colonies were studied. While ST cells multiplied intracellularly and maintained their coccobacillary form in a human macrophage model of infection, SO cells formed long filaments and completely destroyed the phagocytes. In BALB/c mice, the ST variant, but not the SO variant, grew efficiently in the spleen, liver and lung. The ST variant was usually more resistant in vitro than the SO variant to drugs, with MIC values for clarithromycin (CLA), azithromycin (AZI), ciprofloxacin, sparfloxacin, amikacin, clofazimine, ethambutol and isoniazid being higher than those of the SO variant. In beige mice infected with the more highly virulent variant ST, CLA and AZI were the most active drugs in terms of viable count reduction in organs and mutant selection. Together, these observations indicate that the ST variant of M. celatum is a virulent form that can be efficiently inhibited in vivo by CLA and AZI. PMID- 11065353 TI - A novel multidrug efflux transporter gene of the major facilitator superfamily from Candida albicans (FLU1) conferring resistance to fluconazole. AB - Azole resistance in Candida albicans can be mediated by several resistance mechanisms. Among these, alterations of the azole target enzyme and the overexpression of multidrug efflux transporter genes are the most frequent. To identify additional putative azole resistance genes in C. albicans, a genomic library from this organism was screened for complementation of fluconazole hypersusceptibility in Saccharomyces cerevisiae YKKB-13 lacking the ABC (ATP binding cassette) transporter gene PDR5. Among the C. albicans genes obtained, a new gene was isolated and named FLU1 (fluconazole resistance). The deduced amino acid sequence of FLU1 showed similarity to CaMDR1 (formerly BEN(r)), a member of the major facilitator superfamily of multidrug efflux transporters. The expression of FLU1 in YKKB-13 mediated not only resistance to fluconazole but also to cycloheximide among the different drugs tested. The disruption of FLU1 in C. albicans had only a slight effect on fluconazole susceptibility; however, it resulted in hypersusceptibility to mycophenolic acid, thus suggesting that this compound could be a substrate for the protein encoded by FLU1. Disruption of FLU1 in a background of C. albicans mutants with deletions in several multidrug efflux transporter genes, including CDR1, CDR2 and CaMDR1, resulted in enhanced susceptibility to several azole derivatives. FLU1 expression did not vary significantly between several pairs of azole-susceptible and azole-resistant C. albicans clinical isolates. Therefore, FLU1 seems not to be required for the development of azole resistance in clinical isolates. PMID- 11065354 TI - A phosphatidylinositol 3-kinase of Candida albicans influences adhesion, filamentous growth and virulence. AB - To determine if cellular functions of the phosphatidylinositol 3-kinase CaVps34p are related to processes governing Candida albicans pathogenicity, both copies of the gene were sequentially disrupted. Homozygous deletion of C. albicans VPS34 resulted in a mutant strain which exhibited defects not only in intracellular vesicle transport processes but also in morphogenesis. The CaVPS34 null mutant was unable to form hyphae on different solid media whilst showing a significantly delayed yeast-to-hyphae transition in liquid media. In addition, the mutant was rendered hypersensitive to temperature and osmotic stresses and had a strongly decreased ability to adhere to mouse fibroblast cells compared to the wild-type strain SC5314. Finally, evidence was obtained that CaVPS34 is essential for pathogenicity of C. albicans as the CaVPS34 null mutant was shown to be avirulent in a mouse model of systemic infection. C. albicans pathogenicity was restored to a near wild-type degree upon reintroduction of CaVPS34 into the chromosome of the null mutant, demonstrating that the observed avirulence corresponded to the loss of CaVPS34. Thus, the results suggest that CaVPS34 may serve as a potential target for antifungal drugs. PMID- 11065355 TI - Intra- and intermolecular events direct the propeptide-mediated maturation of the Candida albicans secreted aspartic proteinase Sap1p. AB - Pathogenic yeasts of the genus Candida secrete aspartic proteinases (Sap) which are synthesized as preproenzymes. Expression of the C. albicans SAP1 gene lacking the propeptide-coding region in the methylotrophic yeast Pichia pastoris does not lead to the secretion of the enzyme into the culture supernatant, but results in an accumulation of recombinant protein in the cell. Co-expression in this system of the unattached propeptide from Sap1p, as well as from other Saps, restored Sap1p secretion. A deletion analysis revealed that only a 12 aa sequence in the propeptide, corresponding to a highly conserved region in all Sap propeptides, was necessary and sufficient to produce a large amount of Sap1p in culture supernatant. No Sap1p was secreted when Sap1p was produced with a propeptide carrying an F to D mutation in the identified 12 aa sequence. However, the simultaneous production of equivalent amounts of Sap1p and His-tagged Sap1p (H(6) Sap1p) with a mutated and a non-mutated propeptide, respectively, led to the secretion of both proteins in a ratio of approximately 1:2. The restoration of Sap1p secretion occurred at the expense of secretion of H(6)-Sap1p since the total activity was comparable to that of strains producing only H(6)-Sap1p with a non-mutated propeptide. In contrast, the proteolytic activity of strains secreting Sap1p and H(6)-Sap1p both with a functional propeptide was twice that of strains producing either Sap1p or H(6)-Sap1p alone, and the two enzymes were found in an equivalent amount in the culture supernatant. Altogether, these results show that the propeptide can only function once and that the maturation of recombinant C. albicans secreted aspartic proteinase Sap1p is directed through a combination of intra- and inter-molecular pathways. PMID- 11065356 TI - Susceptibility of calves to challenge with Salmonella typhimurium 4/74 and derivatives harbouring mutations in htrA or purE. AB - Salmonella typhimurium 4/74 is highly virulent for cattle after oral challenge, causing severe diarrhoea, which is sometimes associated with systemic spread of the micro-organism. Although susceptible to oral challenge, groups of cattle were found to be relatively resistant to subcutaneous challenge with this strain. The virulence of S. typhimurium 4/74 harbouring mutations in htrA and purE was also assessed in cattle. Although S. typhimurium 4/74 htrA and purE are attenuated following oral challenge in mice, cattle were highly susceptible to oral challenge with these mutants. As with the parent S. typhimurium 4/74 strain, cattle exhibited greater susceptibility to oral compared to subcutaneous challenge with S. typhimurium htrA and purE mutants. Following subcutaneous challenge with sublethal levels of S. typhimurium 4/74, calves produced significant levels of antibodies to S. typhimurium soluble extract. No correlation was detected between interferon gamma levels in sera and susceptibility to infection by any route. The concentrations of the acute-phase associated protein haptoglobin were increased in the sera of five of six cattle inoculated subcutaneously, although increases in concentration were smaller in cattle inoculated orally. PMID- 11065357 TI - Mitogenic factor (MF) is the major DNase of serotype M89 Streptococcus pyogenes. AB - To investigate the role of mitogenic factor (MF) in streptococcal pathogenesis, the structural gene (mf) encoding this protein was disrupted in a clinical isolate of Streptococcus pyogenes H293, to yield the isogenic mutant H363. Growth in enriched broth and on blood agar was unaffected by disruption of mf. Cell-free broth supernatants from H293 and H363 demonstrated identical promitogenic activities when co-incubated with human peripheral blood mononuclear cells, even when diluted 100000-fold, showing that MF is not a major streptococcal mitogen compared with other secreted superantigens. Disruption of mf resulted in complete loss of DNase B production and detectable DNase activity in H363 compared with the parent strain, confirming that the single gene mf, which is present in all group A streptococcal M serotypes studied, encodes DNase B. Despite loss of DNase activity, the virulence of S. pyogenes in a mouse model of necrotizing fasciitis and myositis was unaffected. PMID- 11065358 TI - The complete cps gene cluster from Streptococcus thermophilus NCFB 2393 involved in the biosynthesis of a new exopolysaccharide. AB - The cpsFGHIJKL genes from the cps cluster of Streptococcus thermophilus NCFB 2393 involved in the biosynthesis of EPS were identified, cloned and nucleotide sequenced. The complete cps cluster is contained on an approximately 11.2 kb chromosomal region which contains 12 ORFs, including the previously cloned cpsABCDE genes. Functions were assigned to some of the predicted gene products on the basis of homology to known sequences as follows: cpsK encodes a protein thought to be involved in the polymerization and export of the polysaccharide; cpsE, cpsF, cpsG, cpsH, cpsI and cpsJ encode putative sugar transferases. Two insertion sequences, IS1193 and ISS1, were identified within and flanking the 3' end of the cps cluster respectively. Analysis of the expression of the cpsE gene in Escherichia coli demonstrated that it encodes a glucose-1-phosphate transferase; the enzyme which catalyses the first step in EPS biosynthesis in S. thermophilus NCFB 2393. PMID- 11065359 TI - Involvement of the rml locus in core oligosaccharide and O polysaccharide assembly in Pseudomonas aeruginosa. AB - L-Rhamnose (L-Rha) is a component of the lipopolysaccharide (LPS) core, several O antigen polysaccharides, and the cell surface surfactant rhamnolipid of Pseudomonas aeruginosa. In this study, four contiguous genes (rmlBDAC) responsible for the synthesis of dTDP-L-Rha in P. aeruginosa have been cloned and characterized. Non-polar chromosomal rmlC mutants were generated in P. aeruginosa strains PAO1 (serotype O5) and PAK (serotype O6) and LPS extracted from the mutants was analysed by SDS-PAGE and Western immunoblotting. rmlC mutants of both serotype O5 and serotype O6 synthesized a truncated core region which was unable to act as an attachment point for either A-band or B-band O antigen. A rmd rmlC PAO1 double mutant (deficient in biosynthesis of both D-Rha and L-Rha) was constructed to facilitate structural analysis of the mutant core region. This strain has an incomplete core oligosaccharide region and does not produce A-band O antigen. These results provide the genetic and structural evidence that L-Rha is the receptor on the P. aeruginosa LPS core for the attachment of O polysaccharides. This is the first report of a genetically defined mutation that affects the synthesis of a single sugar in the core oligosaccharide region of P. aeruginosa LPS, and provides further insight into the mechanisms of LPS biosynthesis and assembly in this bacterium. PMID- 11065360 TI - Xenorhabdus bovienii T228 phase variation and virulence are independent of RecA function. AB - Colony pleomorphism, or phase variation, expressed by entomopathogenic bacteria belonging to the genus Xenorhabdus, is an important factor which determines the association of the bacteria with their nematode symbiont and the outcome of infection of susceptible insect larvae by the bacterium- nematode parasitic complex. The mechanism underlying phase variation is unknown. To determine whether RecA-mediated processes are linked to phase variation, the recA gene of Xenorhabdus bovienii was cloned and sequenced. When expressed in a recA-deleted strain of Escherichia coli, the X. bovienii recA clone was able to complement the loss of RecA function. X. bovienii chromosomal recA insertion mutants showed increased sensitivity to UV. Phase 1 forms did not show altered ability to convert to phase 2 and no significant differences in expression of other phase dependent characteristics, including phospholipase C, haemolysin, protease, antibiotic activity and Congo Red binding, were noted. Furthermore, the LD(50) of the X. bovienii recA insertion mutant for Galleria mellonella larvae was not significantly different from that of wild-type strains. From these data the authors conclude that recA is unlikely to be involved in phase variation, the expression of phase-dependent characteristics, or virulence factors involved in killing of susceptible larvae. PMID- 11065361 TI - The plcR regulon is involved in the opportunistic properties of Bacillus thuringiensis and Bacillus cereus in mice and insects. AB - Bacillus thuringiensis has been widely used for 40 years as a safe biopesticide for controlling agricultural pests and mosquitoes because it produces insecticidal crystal proteins. However, spores have also been shown to contribute to overall entomopathogenicity. Here, the opportunistic properties of acrystalliferous B. thuringiensis Cry(-) and Bacillus cereus strains were investigated in an insect species, Galleria mellonella, and in a mammal, BALB/c mice. In both animal models, the pathogenicity of the two bacterial species was similar. Mutant strains were constructed in which the plcR gene, encoding a pleiotropic regulator of extracellular factors, was disrupted. In larvae, co ingestion of 10(6) spores of the parental strain with a sublethal concentration of Cry1C toxin caused 70% mortality whereas only 7% mortality was recorded if spores of the DeltaplcR mutant strain were used. In mice, nasal instillation of 10(8) spores of the parental strain caused 100% mortality whereas instillation with the same number of DeltaplcR strain spores caused much lower or no mortality. Similar effects were obtained if vegetative cells were used instead of spores. The cause of death is unknown and is unlikely to be due to actual growth of the bacteria in mice. The lesions caused by B. thuringiensis supernatant in infected mice suggested that haemolytic toxins were involved. The cytolytic properties of strains of B. thuringiensis and B. cereus, using sheep, horse and human erythrocytes and G. mellonella haemocytes, were therefore investigated. The level of cytolytic activity is highly reduced in DeltaplcR strains. Together, the results indicate that the pathogenicity of B. thuringiensis strain 407 and B. cereus strain ATCC 14579 is controlled by PlcR. PMID- 11065362 TI - Mutational and hyperexpression-induced disruption of bipolar budding in yeast. AB - Analysis of bud-site selection in the yeast Saccharomyces cerevisiae has helped to identify many genes that are generally important for eukaryotic cell polarization. Colony morphology screens were used to identify factors relevant to the process of bipolar budding in yeast. Mutants defective in bipolar budding were identified by virtue of their inability to grow as pseudohyphae in a haploid bud3 background. A mutant allele of the MYO2 gene, encoding a class-V unconventional myosin was identified that perturbs bipolar budding without affecting axial budding and without grossly affecting the role of Myo2p in secretion and maintenance of the actin cytoskeleton. Several genes were also identified whose products, when overexpressed, are capable of disrupting bipolar budding. Among these are the actin-monomer-binding protein profilin and the Aip3p/Bud6p-interacting protein Atc1p. The results strongly support involvement of the actin cytoskeleton in the establishment of bipolar budding and in the maintenance of pseudohyphal growth. PMID- 11065363 TI - Comparative sequence analyses reveal frequent occurrence of short segments containing an abnormally high number of non-random base variations in bacterial rRNA genes. AB - rRNA genes are thought unlikely to be laterally transferred, because rRNA must coevolve with a large number of cellular components to form the highly sophisticated translation apparatus and perform protein synthesis. In this paper, the authors first hypothesized that lateral gene transfer (LGT) might occur to rRNA genes via replacement of gene segments encoding individual domains of rRNA: the 'simplified complexity hypothesis'. Comparative sequence analyses of the 16S and 23S rRNA genes from a large number of actinomycete species frequently identified rRNA genes containing short segments with an abnormally high number of non-random base variations. These variations were nearly always characterized by complementing covariations of several paired bases within the stem of a hairpin. The nature of these base variations is not consistent with random mutations but satisfies well the predictions of the 'simplified complexity hypothesis'. The most parsimonious explanation for this phenomenon is the lateral transfer of rRNA gene segments between different bacterial species. This mode of LGT may create mosaic rRNA genes and occur repeatedly in different regions of a gene, gradually destroying the evolutionary history recorded in the nucleotide sequence. PMID- 11065364 TI - Definition of the attI1 site of class 1 integrons. AB - Integron-encoded integrases recognize two distinct types of recombination site: attI sites, found in integrons, and members of the 59-base element (59-be) family, found in the integron-associated gene cassettes. The class 1 integron integrase, IntI1, catalyses recombination between attI1 and a 59-be, two 59-be, or two attI1 sites, but events involving two attI1 sites are less efficient than the reactions in which a 59-be participates. The full attI1 site is required for high-efficiency recombination with a 59-be site. It is 65 bp in length and includes a simple site, consisting of a pair of inversely oriented IntI1-binding domains, together with two further directly oriented IntI1-binding sites designated strong and weak. However, a smaller region that contains only the simple site is sufficient to support a lower level of recombination with a complete attI1 partner and the features that determine the orientation of attI1 reside within this region. An unusual reaction between the attI1 site and a 59-be appears to be responsible for the loss of the central region of a 59-be to create a potential fusion of two adjacent gene cassettes. PMID- 11065365 TI - Allele-specific PCR shows that genetic exchange occurs among genetically diverse Nodularia (cyanobacteria) filaments in the Baltic Sea. AB - Some cyanobacteria have been shown to exchange genetic information under laboratory conditions, but it has not been clear whether such genetic exchange occurs in the natural environment. To address this, a population genetic study was carried out on the filamentous diazotrophic cyanobacterium Nodularia in the Baltic Sea. Nodularia filaments were collected from 20 widely distributed sampling stations in the Baltic Sea during June and July 1998. Allele-specific PCR (AS-PCR) was used to characterize over 2000 filaments at three loci: a non coding spacer between adjacent copies of the main structural gas vesicle gene gvpA (gvpA-IGS), the phycocyanin intergenic spacer (PC-IGS) and the rDNA internal transcribed spacer (rDNA-ITS). The three loci were all found to be polymorphic in the 1998 population: two alternative alleles were distinguished at the gvpA-IGS and PC-IGS loci, and three at the rDNA-ITS locus. All 12 possible combinations of alleles were found in the filaments studied, but some were much more common than others. The index of association (I:(A)) for all possible pairwise combinations of isolates was found to differ significantly from zero, which implies that there is some linkage disequilibrium between loci. The I:(A) values for 16 out of 20 individual sampling stations also differed significantly from zero: this shows that the observed linkage disequilibrium is not due to pooling data from genetically distinct subpopulations. Monte-Carlo simulations with random subsets of the data confirmed that some combinations showed significantly more linkage disequilibrium than expected by chance alone. It is concluded that genetic exchange occurs in the natural Nodularia population, but the frequency is not high enough for the loci to be in linkage equilibrium. The distribution of the 12 genotypes across the Baltic Sea was found to be non-random, but did not correlate with temperature, salinity or major nutrient concentrations. A significant relationship was found between the gene diversity among filaments at each station and the distance of the station from the centre of the sampling area: possible reasons for this trend are discussed. PMID- 11065366 TI - Stress responses of Synechocystis sp. strain PCC 6803 mutants impaired in genes encoding putative alternative sigma factors. AB - In the complete genome sequence of the cyanobacterium SYNECHOCYSTIS: sp. strain PCC 6803 [Kaneko et al. (1996 ). DNA Res 3, 109-136] genes were identified encoding putative group 3 sigma-factors SigH (Sll-0856), SigG (Slr-1545) and SigF (Slr-1564) and the regulatory protein RsbU (Slr-2031). Mutations in these genes were generated by interposon mutagenesis to study their importance in stress acclimation. For the genes sigH, sigF and rsbU, the loci segregated completely. However, attempts to mutagenize the sigG locus resulted in merodiploids. Under standard growth conditions only minor differences were detected between the mutants and wild-type. However, cells of the RsbU mutant showed a clear defect in regenerating growth after a nitrogen- and sulphur-starvation-induced stationary phase. After applying salt, heat and high-light shocks, stress protein synthesis was analysed by means of one- and two-dimensional electrophoresis. Cells of the SigF mutant showed a severe defect in the induction of salt stress proteins. Although the acclimation to moderate salt stress up to 684 mM NaCl was not significantly changed in this mutant, its ability to acclimate to higher concentrations of NaCl was reduced. Northern blot experiments showed a constitutive expression of the rsbU and sigF genes. The expression of the sigH gene was found to be stress-stimulated, particularly in heat-shocked cells, whilst that of sigG was transiently decreased under stress conditions. Possible functions of these regulatory proteins in stress acclimation of Synechocystis cells are discussed. PMID- 11065367 TI - The SOS promoter dinH is essential for ftsK transcription during cell division. AB - The formation of the Escherichia coli division septum has been well characterized and the majority of the genes involved have been shown to map to the dcw cluster. One exception is ftsK, which lies at 20 minutes, immediately downstream of the global response regulatory gene, lrp. The promoter for ftsK has not yet been assigned. Here, it is reported that ftsK is transcribed from two promoters; the first is located within the lrp reading frame and is dispensable whilst the second is essential and corresponds to dinH, previously characterized as an SOS promoter regulated by LexA. ftsK is the first essential gene to be described that is controlled by an SOS-inducible promoter. A possible mechanism by which dinH may be activated in recA minus strains, or in strains with uncleavable LexA, is discussed. PMID- 11065368 TI - Catabolite repression of dra-nupC-pdp operon expression in Bacillus subtilis. AB - Expression of the Bacillus subtilis dra-nupC-pdp operon is subject to catabolite repression by glucose. It was shown that a cis-acting catabolite-responsive element (CRE) sequence located 64 bp downstream of the transcription-start site mediated catabolite repression of the dra-nupC-pdp operon as it does for many other B. subtilis genes. Point mutations in the CRE sequence caused the loss of catabolite repression of the operon. Catabolite repression of dra-nupC-pdp expression was relieved in a ccpA mutant and was found to be dependent on both HPr and the HPr-like protein Crh. Furthermore, a transcription-repair coupling factor, Mfd, was also found to be involved in the glucose repression of dra-nupC pdp expression. By the use of in vitro gel mobility shift analysis, a specific HPr-P dependent binding of CcpA to the dra CRE site was demonstrated. PMID- 11065369 TI - GcvA binding site 1 in the gcvTHP promoter of Escherichia coli is required for GcvA-mediated repression but not for GcvA-mediated activation. AB - GcvA binds to three sites in the gcvTHP control region, from base -34 to -69 (site 1), from base -214 to -241 (site 2) and from base -242 to -271 (site 3). Previous results suggested that sites 3 and 2 are required for both GcvA dependent activation and repression of a gcvT::lacZ fusion. However, the results were less clear as to the role of site 1. To determine the role of site 1 in regulation, single and multiple base changes were made in site 1 and tested for their ability to alter GcvA-mediated activation and GcvA/GcvR-mediated repression. Several of the mutants were also tested for effects on GcvA binding to site 1 and the ability of GcvA to bend DNA at site 1. The results are consistent with site 1 playing primarily a role in negative regulation of the gcvTHP operon. PMID- 11065370 TI - Identification of DNA-binding proteins involved in regulation of expression of the Streptomyces aureofaciens sigF gene, which encodes sporulation sigma factor sigma(F). AB - Expression of the sigF gene encoding a sporulation-specific sigma factor, sigma(F), in Streptomyces aureofaciens is restricted only to sporulation. Gel mobility-shift assays using protein fractions from different developmental stages of S. aureofaciens revealed two different putative proteins specifically bound to the sigF promoter region: a protein (designated RsfA) present in young substrate mycelium, and a protein (designated RsfB) present in the course of sporulation. Based on the characteristic profiles of their appearance during differentiation, RsfA might be a repressor and RsfB an activator of sigF expression. The location of a specific binding site of the repressor-like protein (RsfA) was determined by gel mobility-shift assays of promoter deletion fragments and by DNase I footprinting analysis. The binding site mapped from nucleotides -87 to -25 relative to the transcription start point of the sigF promoter, and overlapped the -35 promoter region. Given the dependence of sigF expression upon whiH, the putative sporulation transcription factor WhiH was overproduced in Escherichia coli and used in the mobility-shift assays with the sigF promoter. However, no specific binding was detected, indicating an indirect dependence of sigF upon whiH. PMID- 11065371 TI - The Bacillus subtilis 168 csn gene encodes a chitosanase with similar properties to a streptomyces enzyme. AB - The Bacillus subtilis 168 csn gene encodes a chitosanase. It was found that transcription of the csn gene was temporally regulated and was not subject to metabolic repression. Chitosanase synthesis was abolished in a csn mutant strain. Csn was overproduced in B. subtilis, partially purified and characterized. The deduced amino acid sequence, K(m), and optimal pH and temperature of the B. subtilis enzyme were closer to those of a chitosanase from Streptomyces sp. N174 than to those of chitosanases from other Bacillus strains. PMID- 11065372 TI - Transcriptional co-regulation of five chitinase genes scattered on the Streptomyces coelicolor A3(2) chromosome. AB - Streptomyces coelicolor A3(2) strain M145 has eight chitinase genes scattered on the chromosome: six genes for family 18 (chiA, B, C, D, E and H) and two for family 19 (chiF and G). In this study, the expression and regulation of these genes were investigated. The transcription of five of the genes (chiA, B, C, D and F) was induced in the presence of colloidal chitin while that of the other three genes (chiE, G and H) was not. The transcripts of the five induced chi genes increased and reached their maximum at 4 h after the addition of colloidal chitin, all showing the same temporal patterns. The induced levels of the transcripts of chiB were significantly lower than those of the other four genes. Dynamic analysis of the transcripts of the chi genes indicated that chiA and chiC were induced more strongly than chiD and chiF. Addition of chitobiose also induced transcription of the chi genes, but significantly earlier than did colloidal chitin. When cells were cultured in the presence of colloidal chitin, an exponential increase of chitobiose concentration in the culture supernatant was observed prior to the induced transcription of the chi genes. This result, together with the immediate effect of chitobiose on the induction, suggests that chitobiose produced from colloidal chitin is involved in the induction of transcription of the chi genes. The transcription of the five chi genes was repressed by glucose. This repression was apparently mediated by the glucose kinase gene glkA. PMID- 11065373 TI - A novel thermostable multidomain 1,4-beta-xylanase from 'Caldibacillus cellulovorans' and effect of its xylan-binding domain on enzyme activity. AB - The nucleotide sequence of the complete xynA gene, encoding a novel multidomain xylanase XynA of 'Caldibacillus cellulovorans', was determined by genomic-walking PCR. The putative XynA comprises an N-terminal domain (D1), recently identified as a xylan-binding domain (XBD), homologous to non-catalytic thermostabilizing domains from other xylanases. D1 is followed by a xylanase catalytic domain (D2) homologous to family 10 glycosyl hydrolases. Downstream of this domain two cellulose-binding domains (CBD), D3 and D4, were found linked via proline threonine (PT)-rich peptides. Both CBDs showed sequence similarity to family IIIb CBDs. Upstream of xynA an incomplete open reading frame was identified, encoding a putative C-terminal CBD homologous to family IIIb CBDs. Two expression plasmids encoding the N-terminal XBD plus the catalytic domain (XynAd1/2) and the xylanase catalytic domain alone (XynAd2) were constructed and the biochemical properties of the recombinant enzymes compared. The absence of the XBD resulted in a decrease in thermostability of the catalytic domain from 70 degrees C (XynAd1/2) to 60 degrees C (XynAd2). Substrate-specificity experiments and analysis of the main products released from xylan hydrolysis indicate that both recombinant enzymes act as endo-1, 4-beta-xylanases, but differ in their ability to cleave small xylooligosaccharides. PMID- 11065374 TI - A periplasmic, alpha-type carbonic anhydrase from Rhodopseudomonas palustris is essential for bicarbonate uptake. AB - Intact cells of the purple non-sulfur bacterium Rhodopseudomonas palustris growing anaerobically, but not aerobically, contain carbonic anhydrase (CA) activity. The native enzyme was purified >2000-fold to apparent homogeneity and found to be a dimer with an estimated molecular mass of 54 kDa and a subunit molecular mass of 27 kDa. The CA gene (acaP) was cloned and its sequence revealed that it was homologous to alpha-type CAs. The upstream region of acaP was fused to the lacZ gene and beta-galactosidase activity was measured under different growth conditions. Acetazolamide inhibited purified CA with an IC(50) in the range of 10(-8) M, and in the culture media concentrations as low as 30 microM inhibited phototrophic growth under anaerobic, light conditions when bicarbonate was used. An acaP::KAN:(r) mutant strain was constructed by insertion of a kanamycin-resistance cassette and showed a growth pattern similar to wild-type cells grown in the presence of CA inhibitor. CO(2) gas supplied as an inorganic carbon source reversed the effect of mutation or acetazolamide. CA activity measurements, fusion and Western blot experiments confirmed that CA is expressed under different anaerobic conditions independently of bicarbonate or CO(2) and that there is no expression under aerobic conditions. PMID- 11065375 TI - Growth inhibition of Escherichia coli by dichloromethane in cells expressing dichloromethane dehalogenase/glutathione S-transferase. AB - Dichloromethane (DCM) dehalogenase converts DCM to formaldehyde via the formation of glutathione metabolites and generates 2 mol HCl per mol DCM metabolized. Growth of Escherichia coli expressing DCM dehalogenase was immediately and severely inhibited during conversion of 0.3 mM DCM. Intracellular pH (pH(i)) rapidly decreased and chloride ions were steadily released into the medium. Bacterial growth resumed after completion of DCM conversion and cell viability was unaffected. At 0.6 mM DCM there was no recovery from growth inhibition in liquid culture due to the build-up of inhibitory concentrations of formaldehyde. DCM turnover stimulated potassium efflux from cells, which was suppressed by glucose. The potassium efflux, therefore, did not contribute to growth inhibition. It was concluded that initial growth inhibition results from lowering of the cytoplasmic pH, but severity of growth inhibition was greater than expected for the change in pH(i). Possible contributors to growth inhibition are discussed. PMID- 11065376 TI - Control of periplasmic nitrate reductase gene expression (napEDABC) from Paracoccus pantotrophus in response to oxygen and carbon substrates. AB - The napEDABC operon of Paracoccus pantotrophus encodes a periplasmic nitrate reductase (NAP), together with electron-transfer components and proteins required for the synthesis of a fully functional enzyme. Previously, it had been shown that high NAP activity was observed when P. pantotrophus was grown aerobically on highly reduced carbon sources such as butyrate or caproate, but not when cultured on more oxidized substrates such as succinate or malate. The enzyme is not present to any extent when the organism is grown anaerobically under denitrifying conditions, regardless of the carbon source. Transcriptional analyses of the nap operon have now identified two initiation sites which were differentially regulated in response to the carbon source, with expression being maximal when cells were grown aerobically with butyrate. Analysis of a P. pantotrophus mutant (M6) deregulated for NAP activity identified a single C-->A transversion in a heptameric inverted-repeat sequence that partially overlapped the proximal promoter. Transcription analysis of this mutant revealed that expression of nap was completely derepressed under all growth conditions examined. Taken together, these findings indicate that nap transcription is negatively regulated during anaerobiosis, such that expression is restricted to aerobic growth, but only when the carbon source is highly reduced. PMID- 11065377 TI - The Rhizobium leguminosarum bv. viciae glnD gene, encoding a uridylyltransferase/uridylyl-removing enzyme, is expressed in the root nodule but is not essential for nitrogen fixation. AB - A Rhizobium leguminosarum bv. viciae VF39 gene (glnD) encoding the uridylyltransferase/uridylyl-removing enzyme, which constitutes the sensory component of the nitrogen regulation (ntr) system, was identified, cloned and characterized. The deduced amino acid sequence contains the conserved active site motif of the nucleotidyltransferase superfamily and is highly homologous to the glnD gene products of other bacterial species. Downstream of the VF39 glnD resides an open reading frame with similarity to the Salmonella typhimurium virulence factor gene mviN. Mutation of the glnD gene abolished the ability to use nitrate as a sole nitrogen source but not glutamine. In addition, neither uridylylation of P(II) nor induction of the ntr-regulated glnII gene (encoding glutamine synthetase II) under ammonium deficiency could be observed in mutant strains. This strongly suggests that glnD mutants harbour a permanently deuridylylated P(II) protein and as a consequence are unable to activate transcription from NtrC-dependent promoters. The glnD gene itself is expressed constitutively, irrespective of the nitrogen content of the medium. A functional GlnD protein is not essential for nitrogen fixation in R. leguminosarum bv. viciae, but in situ detection of glnD expression in the symbiotic and infection zone of the root nodule and quantitative measurements suggest that at least part of the ntr system functions in symbiosis. The results also indicate that the N terminal part of GlnD is essential for the cell, as deletions in the 5'-region of the gene appear to be lethal and mutations possibly affecting the expression of the first half of the protein have a significant effect on the vitality of the mutant strain. PMID- 11065378 TI - Phylogenetic diversity of rhizobial strains nodulating Robinia pseudoacacia L. AB - Lack of knowledge exists regarding the diversity of rhizobial strains nodulating black locust (Robinia pseudoacacia L.), which is a neophytic tree species widely distributed in Europe. Seventeen rhizobial strains isolated from nodules of black locust at a German location were examined by phenotypic characterization and 16S rDNA analysis. The isolates were classified in nine 16S rDNA genotypes using a set of seven endonucleases. Based on RFLP analysis and sequencing, the strains were shown to belong to the genera Mesorhizobium (76%) and Rhizobium (24%). Five genotypes were identical to the species Mesorhizobium amorphae, Mesorhizobium loti, Mesorhizobium huakuii, Rhizobium leguminosarum and Rhizobium tropici. A strong similarity between the 16S rDNA sequence of another two genotypes and M. amorphae (99.9%) as well as the Mesorhizobium strain R88b (99.8%) was found. The two remaining genotypes were classified in the genus Rhizobium, without a significant relationship at the species level. Comparing isolates nodulating Rob. pseudoacacia and Amorpha fruticosa, a parallel picture of phylogenetic diversity was detected with a range of phylogenetically different rhizobia and M. amorphae dominating. For this study, 18 rhizobial strains which had originally been isolated from a forest in Maryland where black locust is native were additionally analysed. Results revealed seven genotypes all belonging to the genus Mesorhizobium, with four genotypes identical to the isolates from the German sampling location. Whereas the genotype identical to M. amorphae dominated within the strains obtained from the German location, the dominance of a genotype identical to M. huakuii was found among the strains from the native location. Summarizing data from both locations, Rob. pseudoacacia was nodulated with various genomic species, most of which belonged to the genus Mesorhizobium. Concerning phenotypic features such as growth rate, pH tolerance or use of certain carbohydrates, most isolates corresponded to described species and genera. However, there were differences in salt tolerance between these isolates and the corresponding reference strains. Overall, the results demonstrated a high phenotypic and phylogenetic diversity of rhizobial strains nodulating Rob. pseudoacacia. This may be a characteristic of neophytic and other widely spread legumes and may contribute to the success of black locust as a pioneer tree species for the temperate zone. PMID- 11065379 TI - Lipid composition and taxonomy of [Pseudomonas] echinoides: transfer to the genus Sphingomonas. AB - Lipid components of [Pseudomonas] echinoides NCIMB 9420 have been studied as an aid to taxonomic relocation of the organism. Non-polar lipids include the carotenoid nostoxanthin and the ubiquinone Q-10. The major fatty acids are cis vaccenic acid [18:1(11c)], hexadecanoic acid (16:0) and 2-hydroxy-tetradecanoic acid (2-OH-14:0), but 11-methyloctadec-11-enoic acid[11-Me-18:1(11)] is a significant minor component. The preponderant phospholipids are phosphatidylethanolamine and phosphatidylglycerol; minor lipids include bis(phosphatidyl)glycerol and an unidentified aminophospholipid. Several glycolipids are present, the major one being a glucuronosylceramide derived from sphinganine with amide-bound 2-OH-14:0. The lipid profile supports a proposal to reclassify the organism as Sphingomonas echinoides. PMID- 11065380 TI - High hopanoid/total lipids ratio in Frankia mycelia is not related to the nitrogen status. AB - Vesicles are specific Frankia structures which are produced under nitrogen limiting culture conditions. Hopanoids are the most abundant lipids in these vesicles and are believed to protect the nitrogenase against oxygen. The amounts and quality of each hopanoid were estimated in different Frankia strains cultivated under nitrogen-depleted and nitrogen-replete conditions in order to detect a possible variation. Studied Frankia strains nodulating Eleagnus were phylogenetically characterized by analysis of the nifD-K intergenic region as closely related to genomic species 4 and 5. Phylogenetically different strains belonging to three infectivity groups were cultivated in the same medium with and without nitrogen source for 10 d before hopanoid content analysis by HPLC. Four hopanoids together accounted for 23-87% and 15-87% of the total lipids under nitrogen-replete and nitrogen-depleted culture conditions, respectively. Two of the hopanoids found, bacteriohopanetetrols and their phenylacetic acid esters, have previously been described in Frankia Two new hopanoids, moretan-29-ol and a bacteriohopanetetrol propionate, have also been identified. The moretan-29-ol and bacteriohopanetetrols were found to be the most abundant hopanoids whereas the bacteriohopanetetrol propionate and phenylacetates were present at a concentration close to the limit of detection. The ratio of (bacteriohopanetetrols + moretan-29-ol)/(total lipids) varied in most of the strains between nitrogen-depleted and nitrogen-replete culture conditions. In most of the strains, the hopanoid content was found to be slightly higher under nitrogen-replete conditions than under nitrogen-depleted conditions. These results suggest that remobilization, rather than neosynthesis of hopanoids, is implicated in vesicle formation in Frankia under nitrogen-depleted conditions. PMID- 11066009 TI - The Human Genome Project: a generation's psyche and a society's revolution. PMID- 11066010 TI - Genetic testing for ataxia in North America. AB - BACKGROUND: The Ataxia Molecular Diagnostics Testing Group was established to generate quantitative proficiency and outcomes data regarding molecular testing for the autosomal dominant cerebellar ataxias (spinocerebellar ataxia types 1 [SCA-1] through -3, -6, and -7, and dentatorubral-pallidoluysian atrophy) in North America. METHODS AND RESULTS: Twenty-four North American laboratories that offered diagnostic testing for one or more ataxia genes were initially identified through GeneTests (Children's Health Care System, Seattle, WA). Eighteen laboratories agreed to participate in the study, which consisted of completing a technical survey, clinical survey, and molecular proficiency test. One laboratory returned the completed surveys but did not perform the proficiency testing. Ten of 18 laboratories (56%) provided data on test volumes, and these laboratories collectively performed 2,240 tests; approximately 5% of the tests yielded a positive result (i.e., identification of a pathological trinucleotide (CAG) repeat expansion). In proficiency testing, 100% of the laboratories correctly genotyped all samples, and 93% of the laboratories were within 1 SD of the mean for sizing normal alleles (one repeat unit or less). Ninety percent of the laboratories were within 1 SD for sizing expanded alleles. CONCLUSIONS: Proficiency testing showed little difference between laboratories with respect to allele sizing. However, additional phenotype/genotype correlations are necessary to define CAG repeat-length descriptors for SCA-1, SCA-2, and SCA-7 alleles of intermediate size. PMID- 11066011 TI - Evaluation of chimerism in DNA samples by PCR amplification of D1S80 with detection by capillary electrophoresis. AB - BACKGROUND: Analysis of the relative amounts of donor and recipient DNA in bone marrow after bone marrow transplantation is frequently used to determine the status of the transplant. We studied the performance of an assay to quantify chimerism based on amplification of the D1S80 variable number tandem repeat marker by PCR with detection of PCR products by capillary electrophoresis (CE). METHODS AND RESULTS: Samples from potential bone marrow donors and recipients were analyzed separately and in mixtures to simulate various degrees of chimerism from 10% to 90% and subjected to PCR/CE analysis. There was excellent agreement between the measured and known relative proportions of DNA components in chimeric samples. The lower limit of sensitivity for detection of chimerism was 1%; between-runs coefficients of variation were <5%. CONCLUSIONS: Amplification of the D1S80 minisatellite by PCR with CE detection is a reliable method for determination of the relative contribution of different DNAs in mixed samples. This method is fast, quantitative, and extremely reproducible. PMID- 11066012 TI - Simultaneous detection of C282Y and H63D hemochromatosis mutations by dual-color probes. AB - BACKGROUND: Hemochromatosis is a common genetic disease, affecting one in every 200 individuals in the United States. A PCR assay was designed using fluorescent melting curve analysis to simultaneously detect the G845-->A (C282Y) and C187-->G (H63D) mutations. The G845-->A and C187-->G loci are distinguished by color, and mutant alleles are distinguished from wild type by probe melting temperature (Tm). METHODS AND RESULTS: The probe sets used two fluorophore pairs, fluorescein with LCRed 640 for G845-->A and fluorescein with LCRed 705 for C187-->G. The probes, complementary to the mutant allele, dissociate from the product at specific Tms. Wild-type alleles form mismatches with the probes, reducing the Tms by 6 degrees C (G845-->A) and 10 degrees C (C187-->G). One of 133 samples had a Tm shift 4 degrees C less than the wild-type Tm for the G845-->A locus. Sequencing confirmed the sample to be homozygous for G845-->A and heterozygous for a C-->A substitution at position 842 (C842-->A), substituting lysine for threonine. CONCLUSIONS: Multiplexing by color and Tm allows for simultaneous genotyping of each mutation. A novel base-pair alteration was detected in cis with a G845-->A mutation. PMID- 11066013 TI - Detection of a novel truncated WT1 transcript in human neoplasia. AB - BACKGROUND: The Wilms' tumor 1 (WT1) gene encodes a transcription factor critical in urogenital development. Using a new model of prostate cancer progression that permits comparison of the cellular and molecular properties of increasingly aggressive sublines of simian virus 40 large T-antigen-immortalized human prostate epithelial cells within the same lineage, the role of WT1 in tumorigenesis was investigated. METHODS AND RESULTS: Using RT-PCR and northern blotting, we identified a novel truncated WT1 transcript in these prostate cancer cell lines. This 2.1-kb transcript consisted of the coding region of the zinc finger domain of WT1, together with a portion of intron 5 at the 5' end of the transcript. Furthermore, two peptides were detected by western blotting using antibodies to epitopes of the COOH terminus of WT1. Using RT-PCR, the 2.1-kb transcript was also detected in leukemia cell line K562, breast cancer cell line MCF7, and blood samples from patients with acute leukemia. CONCLUSION: These novel findings in both cell lines and patient-derived specimens suggest this new WT1 gene alteration has a potential role in the development of new diagnostic assays for some human malignancies. PMID- 11066014 TI - Quantitative evaluation of post-bone marrow transplant engraftment status using fluorescent-labeled variable number of tandem repeats. AB - BACKGROUND: The analysis of highly polymorphic variable number of tandem repeat (VNTR) loci is useful for the estimation of donor-host chimerism in bone marrow transplant recipients. METHODS AND RESULTS: A rapid and sensitive engraftment assay has been developed in which the VNTR loci, D1S80, D17S5, D1S111, and apoB, are amplified with fluorescent-labeled (Cy5.5) oligonucleotide primers, followed by analysis using the Visible Genetics, Inc, OpenGene System. The degree of chimerism is then calculated by determining the percentage of host contribution to the total informative allele peak area. Reconstitution experiments and analysis of 383 posttransplantation DNA samples, isolated from 71 different bone marrow transplant recipients, were evaluated as part of assay development. Reconstitution studies showed assay linearity and sensitivity of at least 1%. Patient results were compared with a previous analysis in which unlabeled PCR products were quantified on silver-stained polyacrylamide gels. High concordance was observed between fluorescent analysis and silver-staining method in all 71 patients. CONCLUSIONS: Fluorescent analysis offers many advantages over previous methods, including faster turnaround time, decreased DNA requirements, greater resolution and/or sensitivity, and objective interpretation. PMID- 11066015 TI - Diagnostic value of PCR for detection of Borrelia burgdorferi DNA in clinical specimens from patients with erythema migrans and Lyme neuroborreliosis. AB - BACKGROUND: The aim of the study is to evaluate the diagnostic sensitivity of a 16S ribosomal RNA-based PCR on clinical specimens from patients with erythema migrans (EM) and neuroborreliosis and to compare the sensitivities with those obtained by in vitro culture and serological testing. A semiquantitative detection system, representing the input amount of specific DNA and thus the density of spirochetes in clinical specimens, indicated the preferred clinical sample to obtain for PCR testing. METHODS AND RESULTS: Skin biopsy and urine samples from 31 patients with EM and cerebrospinal fluid (CSF) and urine samples from 30 patients with neuroborreliosis were investigated. Borrelia burgdorferi DNA was detected in 71% of the skin biopsy specimens and 13% of the urine samples from patients with EM. Forty-one percent of the patients with EM were found to have B burgdorferi-specific antibodies in serum, and B burgdorferi was cultured in 29% of the EM specimens. For patients with neuroborreliosis, the diagnostic sensitivities in CSF and urine samples were 17% and 7%, respectively. Specific intrathecal antibody production was found in 90% of the patients, and 87% showed elevated B burgdorferi antibodies in serum. In general, PCR of skin biopsy samples yielded very high amounts of amplicons versus low amounts for CSF and urine samples. CONCLUSIONS: PCR of skin biopsy specimens is currently the most sensitive and specific test for the diagnosis of patients with EM, superior to culture and serological testing. For B burgdorferi-specific CSF disgnosis in patients with neuroborreliosis, the measurement of specific intrathecal antibody synthesis is superior to PCR. However, in patients with a short duration of disease (<14 days), PCR may be a useful diagnostic supplement. PCR of urine samples cannot be recommended at the present time for routine diagnosis of patients with EM or neuroborreliosis. PMID- 11066016 TI - Efficient recovery of DNA from peripheral blood for diagnostic analysis with a vacuum manifold. AB - BACKGROUND: The increasing clinical use of diagnostic DNA mutation analysis requires efficient isolation of DNA from peripheral blood. METHODS AND RESULTS: The use of a vacuum manifold to isolate DNA was evaluated and compared with a similar centrifugation-based DNA isolation technique. In PCR-based assays of five point mutations, identical results were obtained with DNA isolated from peripheral blood using either centrifugation or a vacuum system. Minor modifications to PCR procedures were encountered. CONCLUSIONS: In the clinical setting, this vacuum-driven method of DNA isolation provides an efficient, useful alternative to conventional centrifugation-based DNA isolation from peripheral blood specimens. Providing sufficient, stable DNA for multiple assays, it is easily implemented without highly specialized, expensive equipment and decreases the time spent isolating DNA from multiple samples. In addition, the potential for specimen contamination is reduced because there are fewer transfer steps. PMID- 11066017 TI - Chronic fatigue syndrome is not associated with expression of endogenous retroviral p15E. PMID- 11066018 TI - Thrombophilic mutations impart a high risk of pregnancy-related venous thrombosis. PMID- 11066019 TI - Characterization of a microprocessor-controlled tubular multiple metered dose inhaler aerosol generator for inhalation exposures of pharmaceuticals. AB - A microprocessor-controlled tubular multiple metered dose inhaler (MDI) aerosol generator was constructed for the delivery of pharmaceutical aerosols to inhalation chambers. The MDIs were mounted in four cassettes containing one to four MDIs on a stepped end plate. The MDIs in each cassette were pneumatically activated at intervals that were controlled by the microprocessor. The cassettes permitted easy replacement of each set of MDIs with a fresh set of MDIs whenever necessary. Aerosol concentration was controlled by varying the number of active MDIs in each cassette and the frequency of activations per minute of each row. Aerosol from the MDIs flowed along the long axis of the tube, which provided a path length sufficient to diminish impaction losses. Using a light-scattering device to monitor the aerosol concentration, the pulsatile output from the MDIs in the cassettes was demonstrated to be adequately damped out provided that the dilution/mixing/aging chamber exceeded 3 ft in length. The tube diameter selected was the minimum compatible with mounting the required number of MDIs so that the linear velocity of the aerosol was adequate to efficiently transport the aerosol out of the dilution chamber. Aerosol concentration and particle size data were recorded for a nose-only rodent exposure chamber. Reproducible aerosol concentrations ranging from 0.03 to 0.6 mg/L were generated. Particle sizes ranged from 2- to 3-microm mass median aerodynamic diameter. Thus, the aerosol generated was within the size range suitable for inhalation exposures. PMID- 11066020 TI - Accounting for radioactivity before and after nebulization of tobramycin to insure accuracy of quantification of lung deposition. AB - The ability to predict drug deposition of inhaled drugs used in cystic fibrosis (CF) is important if there is a need to target specific doses of drug to the lungs of individual patients. The gold standard of measuring pulmonary deposition is the quantification of an aerosolized radiolabel either mixed with the drug solution or tagged directly to the compound of interest. Accuracy of the quantification could be assured if there is agreement between the amount of radioactivity before and after administration. Before administration, the radiolabel is concentrated in the well of the nebulizer, whereas after administration, it is distributed throughout the nebulizer, the expiratory filter and connectors, and the upper airway, stomach, trachea, and lung. Not only is the geometry of the distribution that is presented to the gamma camera different, but there are different attenuation factors for the various body tissues. The primary aim of this study was to evaluate the accuracy of the quantification of deposition. Secondary goals were to compare in vitro nebulizer performance with that measured in vivo during the deposition study. Eighty milligrams of tobramycin and technetium bound to human serum albumin was administered to 10 normal adults using a Pari LC Jet Plus (Pari Respiratory Equipment, Inc., Richmond, VA) breath-enhanced nebulizer. Techniques were developed that allowed for the accounting of 99 +/- 2% of the initial radioactivity. The fraction of the rate of lung deposition to total body deposition was the in vivo respirable fraction (0.62 +/- 0.07), which closely agreed with in vitro measurements of respirable fraction (0.62 +/- 0.04). Drug output measured from the change in weight and concentration in the nebulizer systematically overestimated drug output measured by the deposition study. The results indicate that 11.8 of the initial 80 mg would be deposited in the lungs. This technique could be adapted to accurately quantify the amount of deposition on any inhaled therapeutic agent, but caution must be used when extrapolating performance of a nebulizer on the bench to expected deposition in patients. PMID- 11066021 TI - An investigation of the solubility of various compounds in the hydrofluoroalkane propellants and possible model liquid propellants. AB - The aims of this study were to investigate descriptive parameters that may predict the solubility of compounds in the hydrofluoroalkane (HFA) propellants and to identify a model HFA propellant that is liquid at room temperature and atmospheric pressure. The solubility of 32 and 20 compounds chosen to give a wide range of physicochemical properties in HFA-134a and HFA-227, respectively, was measured. The Fedors solubility parameter and a computed log octanol water partition coefficient (CLOGP) were compared with the compounds' solubility in the HFA propellants. A total of 19 and 15 solutes had finite solubilities for HFA 134a and HFA-227, respectively, although the remaining solutes were miscible in all proportions. There was no apparent relation between solubility in HFA and the Fedors solubility parameter. This was not improved by considering the hydrogen bonding potential of the compounds. When log solubility versus CLOGP was plotted, there was a linear relation for 16 and 12 of the compounds exhibiting a finite solubility in the HFA propellants, although four solutes (phenols) were displaced to the left of the linear relation. The remaining 3 compounds had much lower solubilities than was predicted from their CLOGPs, possibly as a consequence of their crystallinity (high melting points). Of the putative model propellants investigated (i.e., perfluorohexane (PFH), 1H-perfluorohexane [1H-PFH], and 2,2,2 trifluoroethanol), 1H-PFH was the most promising, with a linear relation between solubility in 1H-PFH and solubility in HFA propellant being observed. The solubilities in 1H-PFH were approximately 11 and 26% of those in HFA-134a and HFA 227. PMID- 11066022 TI - Evaluation of the accuracy and precision of lung aerosol deposition measurements from single-photon emission computed tomography using simulation. AB - Single-photon emission computed tomography (SPECT) imaging is being increasingly used to assess inhaled aerosol deposition. This study uses simulation to evaluate the errors involved in such measurements and to compare them with those from conventional planar imaging. SPECT images of known theoretical distributions of radioaerosol in the lung have been simulated using lung models derived from magnetic resonance studies in human subjects. Total lung activity was evaluated from the simulated images. A spherical transform of the lung distributions was performed, and the absolute penetration index (PI) and a relative value expressed as a fraction of that in a simulated ventilation image were calculated. All parameters were compared with the true value used in the simulation, and the errors were assessed. An iterative method was used to correct for the partial volume effect, and its effectiveness in improving errors was evaluated. The errors were compared with those of planar imaging. The precision of measurements was significantly better for SPECT than planar imaging (2.8 vs 6.3% for total lung activity, 6 vs 20% for PI, and 3 vs 6% for relative PI). The method of correcting for the influence of the partial volume effect significantly improved the accuracy of PI evaluation without affecting precision. SPECT is capable of accurate and precise measurements of aerosol distribution in the lung, which are improved compared with those measured by conventional planar imaging. A technique for correcting the SPECT data for the influence of the partial volume effect has been described. Simulation is demonstrated as a valuable method of technique evaluation and comparison. PMID- 11066023 TI - A system to reproduce human breathing patterns: its development and validation. AB - Studies of aerosol deposition in models of the human respiratory tract play a significant role in developing our understanding of drug delivery by inhalation and particle retention in the lungs during exposure to polluted environments. To use replica casts of human airways and compare the results with in vivo data, a device is required to simulate human breathing. The objective of this study was to simulate human breathing for nasal casts. Breathing through the nose is normally limited to about 50 L/min. Therefore, a system was built to simulate human breathing patterns as well as artificial ones up to this flow rate. The system consists of a reciprocating piston in a cylinder, which is displaced by a synchronous motor via a linear actuator. The desired signal to drive the motor is given in real time by purpose-written software. The rotation and position of the motor are controlled by an electronic position control unit. The validation of the system shows that it simulates breathing up to 50 L/min closely even for complex waveforms. At breathing rates above 50 L/min, a slight difference is apparent between the desired breathing pattern and the simulated one. The breathing simulator has been shown to be a reliable tool for reproducing a wide variety of breathing patterns. PMID- 11066024 TI - Regulation of differentiation of the tracheobronchial epithelium. AB - The study of differentiation has been the domain of embryologists and developmental biologists and, in the pulmonary field, the concern of neonatologists. Why should those of us who are neither be interested in differentiation of the epithelium lining the conducting airways? The reason is that injury to the airway epithelium and disruption of its steady state and its normal differentiation are common occurrences in both acute episodes of infection and during chronic diseases such as chronic obstructive pulmonary disease and asthma. Thus, it is important to know how injury is repaired and which are the critical mechanisms that control and regulate differentiation. PMID- 11066025 TI - Lack of nitric oxide involvement in cholinergic modulation of ovine ciliary beat frequency. AB - Ciliary beat frequency (CBF) is regulated, at least in part, by the cytoplasmic calcium concentration ([Ca(2+)](i)). Because Ca(2+) can stimulate nitric oxide (NO) production by nitric oxide synthase (NOS) and NO has been implicated in the regulation of CBF in some species, we examined whether NOS is present in cultured ovine ciliated epithelial cells and whether NO plays a role in the Ca(2+) mediated muscarinic stimulation of CBF. Dissociated ovine tracheal epithelial cells were grown in culture for 2 to 14 days. Frequency from a single cilium was measured by on-line Fourier transform methods using video microscopy. [Ca(2+)](i) was determined with fura-2 using fluorescence ratio imaging from the same single cells. Ciliated cells contained NOS in culture as indicated by NADPH-diaphorase staining. Acetylcholine (ACh) increased CBF and [Ca(2+)](i) transiently as previously shown. Measurements with 2',7'-dichlorofluorescin diacetate indicated that reactive oxygen/nitrogen species were produced in these cells on ACh exposure. NOS inhibitors N(G)-nitro-L-arginine methyl ester (< or =10 mM), N(G) nitro-L-arginine (< or =10 mM), and 7-nitro indazole (1 microM) were unable to block the CBF or [Ca(2+)](i) response to ACh. Furthermore, the NO donors sodium nitroprusside and S-nitroso-N-acetylpenicillamine (< or =1 mM) did not change CBF or [Ca(2+)](i). Above these concentrations, they both lead to a reversible decrease in CBF. The membrane-permeable cyclic guanosine monophosphate analogue 8 bromo-cyclic guanosine monophosphate had no effect on CBF, whereas 8-bromo-cyclic adenosine monophosphate stimulated CBF. Taken together, these results suggest that NO does not play a role in mediating the ACh-induced increase in CBF through [Ca(2+)](i). The role and targets for NO in ovine ciliated cells remains to be determined. PMID- 11066026 TI - Hyaluronic acid in cultured ovine tracheal cells and its effect on ciliary beat frequency in vitro. AB - Hyaluronic acid (hyaluronan, or HA) is secreted by submucosal glands, but its function in airway secretions other than influencing the rheology of mucus is not fully understood. HA is known to modulate cell behavior and to enhance sperm motility. Because sperm tails and cilia have the same microtubular structure, we studied the effect of HA on ciliary beat frequency (CBF) in vitro. CBF of cultured ovine airway epithelial cells was measured continuously by digital video microscopy. After removal of endogenous HA by hyaluronidase, cells were exposed to 50 to 100 microg/mL of HA at different times in culture. No change in CBF in response to HA was seen in cells cultured less than 7 days. After 7 days, however, 6 of 10 measured cells (from three different sheep) showed a transient CBF increase from a baseline of 6.4 +/- 0.3 Hz (mean +/- SE) to 7.4 +/- 0.4 Hz or 16% above baseline (p < 0.05). At these time points (but not before), cytochemical staining was positive for endogenous HA using a biotinylated HA binding protein. These data suggest that HA can increase CBF of tracheal epithelial cells only late in culture when HA is able to bind to an unspecified cell surface structure. Because this binding has a physiological effect, we hypothesize that it is an HA-binding receptor, that is either transiently expressed late in culture or initially destroyed by the protease treatment for cell dispersion. PMID- 11066027 TI - Impairment of airway mucociliary transport in patients with sinobronchial syndrome: role of nitric oxide. AB - Nitric oxide (NO) produced within the respiratory tract can stimulate ciliary motility of airway epithelial cells and hence mucociliary transport. In this study, to determine the role of NO in mucociliary dysfunction in sinobronchial syndrome (SBS), we measured NO concentrations in the exhaled air by chemiluminescence analyzer and nasal clearance time (NCT) by saccharin test. Exhaled NO concentrations in patients with SBS were 39% of those in healthy nonsmokers and 55% of those in healthy smokers. The patients also showed prolonged NCT compared with healthy subjects, and there was a significant negative correlation between exhaled NO concentrations and NCT. Furthermore, concentrations of chloride (Cl) in the sputum supernatant were higher in SBS patients than in healthy subjects, and there was a significant negative correlation between sputum Cl concentrations and exhaled NO concentrations. These results suggest that airway mucociliary clearance is impaired in patients with SBS and that this impairment might result from the reduced production of NO and the impaired availability of the molecule in the mucociliary apparatus. PMID- 11066028 TI - Model systems for investigating mucin gene expression in airway diseases. AB - Overproduction of mucus and of mucin glycoproteins and goblet cell hyperplasia occurs in chronic obstructive airway diseases, including asthma and cystic fibrosis. Mucus overproduction results from alterations in several cellular processes, including altered regulation of airway mucin genes on exposure to environmental and infectious agents and to inflammatory mediators. Seven of the nine identified MUC genes (which encode the protein backbone of mucins) are normally expressed in human respiratory tract tissues. Several inflammatory mediators have now been shown to regulate expression of MUC2, MUC5AC, and MUC5B genes. Importantly, mucin gene expression can be regulated both transcriptionally and posttranscriptionally. Current information on airway mucin gene expression is summarized in this review along with an overview of airway epithelial model systems. In vitro model systems include airway epithelial carcinoma cell lines and primary normal human bronchial epithelial (NHBE) cells. In vivo systems include human respiratory tract tissues and rodent airways. Our laboratory has begun to investigate the role of cytokines on mucin gene expression in vitro and in vivo and on goblet cell metaplasia in vivo. Because cytokines can alter cell proliferation, we characterized the effect of interleukin (IL)-4 and IL-13 on the proliferation of NHBE cells and three human lung carcinoma cell lines--A549, NCI H292, and Calu-3--that are frequently used for analyses of airway mucin gene expression. Both IL-4 and IL-13 had cell-specific effects. They increased proliferation moderately (1.2-3.0-fold) in NHBE and Calu-3 cells, but markedly inhibited proliferation of A549 cells in a dose-dependent manner. IL-4 increased proliferation of NCI-H292 cells moderately, although IL-13 had no significant effect. We also examined the role of IL-13 and IL-4 on MUC5AC messenger RNA (mRNA) expression in A549, Calu-3, and H292 cell lines and did not observe any significant effect. However, we recently showed an increase in Muc-5ac mRNA and protein expression in a murine model of ovalbumin-induced allergic asthma and in murine airways when IL-13 was delivered intranasally (Alimam, N.Z., et al. Am J. Respir. Cell Mol. Biol. 22:253--260). Thus, we speculate that IL-13 plays a role in the differentiation of murine airway epithelial cells into goblet cells, which then express Muc-5ac mRNA. A detailed analysis of the role of cytokines in airway cell differentiation and mucin gene expression both in vitro and in vivo is required to elucidate the roles of mucins in airway health and diseases. Identification of Muc-5ac as a major gene and gene product in goblet cell metaplasia should facilitate delineation of the molecular mechanisms underlying the induction and reversal of airway goblet cell metaplasia and goblet cell hyperplasia. PMID- 11066029 TI - Respiratory-related quality of life: relation to pulmonary function, functional exercise capacity, and sputum biophysical properties. AB - One of the difficulties in assessing mucoactive therapy is selecting clinical outcome variables that reflect the impact of clearing airway secretions on quality of life (QOL). Petty and colleagues developed a questionnaire designed to evaluate the clinical impact of mucoactive therapy in patients with chronic bronchitis (CB). We evaluated this questionnaire in a multicenter study of a mucolytic medication used in patients with CB and hypothesized that spirometry, exercise capacity, and sputum clearability changes would correlate with QOL changes. This was a multicenter trial in 159 patients with stable CB (111 completed the 16-week study). Spirometry, plethysmography, the 6-minute walk test (6MWT), and Petty score as a measure of QOL were assessed at each visit. Sputum was collected at each visit. Cough transportability was measured in a cough machine, and mucociliary transportability was measured on the frog palate. Cohesivity was measured in a filancemeter, interfacial tension by de Nouy ring, and wettability by contact angle analysis. Within the entire data set of 694 evaluations, there was no correlation between pulmonary function and QOL. There was an inverse correlation with distance covered in a 6MWT (R(2) = 0.041, p < 0.0001). Sputum CTR was directly correlated with QOL (R(2) = 0.027, p < 0.0001). Change from baseline (mean of first three visits) was computed and compared the change in the mean of values at the 8- and 12-week visits (n = 108 sets of data pairs). This was analyzed as a percentage of change for continuous measurements, and as QOL is normative, we calculated the absolute change in QOL. There was no relation between QOL and 6MWT changes. There was an inverse relation between change in forced expiratory volume in 1 second and QOL (R(2) = 0.092, p = 0.0021) as well as between forced vital capacity and QOL (R(2) = 0.05, p = 0.024). There was a direct relation between CTR and QOL (R(2) = 0.039, p = 0.048). The relation between QOL and 6-minute walk distance was expected but weak. The consistent relation between CTR and QOL (suggesting that improved CTR of sputum is associated with decreased QOL) is difficult to explain. A change in forced expiratory volume in 1 second and forced vital capacity did correlate with a change in QOL. There is a need for a good QOL tool to evaluate mucus clearance devices or medications. The Petty questionnaire was designed specifically for this task, but the effect on sputum properties by current mucoactive agents may be too small to elicit a significant change in the Petty score. PMID- 11066030 TI - Protein kinase C regulates the flow rate-dependent decline in human nasal ciliary beat frequency in vitro. AB - Cilia provide the driving force for mucociliary clearance, the process that removes mucus from the airways. Protein kinase C (PKC) plays a poorly understood regulatory role in phosphorylation-based signal transduction cascades, including the control of human mucociliary clearance, especially with respect to ciliary beat frequency (CBF). Ciliary studies minimize the importance of fluid flow, because it is generally accepted that flow increases CBF. Here, we studied postflow events by measuring CBF in vitro in volunteers. Rose chamber-loaded cells were pulsed for 5 minutes at 30 mL/h in medium-199 +/- PKC modulators at 20 degrees C. The 5-minute pulse precipitated a fall in CBF noted within 1 minute after flow (acute dip response [ADR] to 84 +/- 2% of preflow baseline). Thereafter, CBF rose to 8% below baseline for 30 minutes [postrecovery plateau at 92 +/- 3%]. Preincubation with 1 microM of phorbol 12-myristate 13-acetate (PMA), a PKC-activating phorbol ester attenuated the ADR (c. 95%) and restored the postrecovery plateau almost to baseline levels (98 +/- 0.7%; p > 0.10 compared with baseline CBF). With respect to the ADR, the PMA protective effect was lost in the presence of the selective PKC inhibitor myristoylated epidermal growth factor peptide 651d-658 (Myr-PKCI; 10 microM). Myr-PKCI alone changed the ADR pattern such that the CBF remained at 15% below preflow baseline. We conclude that CBF fall and recovery after a fluid pulse is regulated by PKC activity either directly or indirectly. PMID- 11066031 TI - Formulation of inhaled medicines: effect of delivery vehicle on immortalized epithelial cells. AB - The small volume of airway lining fluid renders it susceptible to alteration by the deposition of inhaled formulations. The increasing popularity of the pulmonary route for drug delivery has led to an increasing number of pharmaceutical excipients being incorporated into inhaled dosage forms. The effects of drug delivery vehicle on airway epithelial cells can be studied with the aid of cell culture models of the respiratory epithelium. The effects of pH, osmolarity, and lactose on epithelial cell layers were studied using 16HBE14o- cells. Mannitol flux was used to assess epithelial permeability, enzymatic conversion of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) was used as a measure of epithelial cell metabolism, and release of the cytosolic enzyme lactate dehydrogenase was used as a measure of cell integrity. The effect of buffer composition on epithelial cell mucus secretion was studied using HT29 clone H cells, with mucus secretion measured using an enzyme-linked lectin assay. The permeability of 16HBE14o- cell layers was increased by apical fluid of pH 5, 6, and 9 as well as osmolarities of 96 and 545 mOsm. MTT conversion was reduced by apical fluid of pH 5 and 6 and osmolarity of 96 mOsm. Lactate dehydrogenase release was only increased by apical fluid of pH 9. No effect of lactose solution (100 mM) on the epithelial cells was observed. Mucus secretion by HT29-clone H cells was lowest in Dulbecco's modified Eagle medium (2.92 +/- 0.23 ng per cell layer) and was increased in phosphate-buffered saline with magnesium and calcium (4.28 +/- 0.38 ng per cell layer) and phosphate-buffered saline without magnesium and calcium (6.56 +/- 0.72 ng per cell layer). These results suggest that the physicochemical properties of inhaled formulations should be carefully controlled. The effect of buffer composition on mucus secretion suggests that even the application of "physiological" solutions may affect the epithelium. These cell models represent an opportunity to investigate the interaction of drug delivery vehicles with the epithelium. PMID- 11066032 TI - Cell proliferation and death in the gastric epithelium of developing rats after glucocorticoid treatments. AB - Glucocorticoids take part in the intense morphofunctional modifications that occur in the gastric mucosa during fetal and postnatal development. Two studies were designed to evaluate corticoids role in gastric cell proliferation and apoptosis in developing rats: in vivo, using suckling animals; in vitro, using gastric explants obtained from 20-day fetuses. These explants were cultured in DMEM/F12 medium treated or not with 50 ng/ml of corticosterone; after 22 hr, vincristine was added to the medium for 2 hr to block mitosis. The metaphasic index decreased significantly after the 24-hr treatment (controls: 1.52 +/- 0.53; treated: 0.40 +/- 0.21) and apoptotic cells were visualized under light and electron microscopy. Fifteen-day-old rats were treated with hydrocortisone (25 mg/Kg) for 3 days, and injected with BrDU (100 mg/Kg) 1 hr before sacrifice on the 18th day. BrDu-labeled and non-labeled cells were counted to determine the labeling index of epithelial cells. As apoptotic cells are rapidly eliminated, other animals were treated for only 2-3 hr. Sections were investigated for the presence of apoptotic cells, using morphological criteria and TUNEL labeling. Hydrocortisone significantly reduced the labeling index (controls: 15.6 +/- 1.6 vs. treated: 11.7 +/- 1.1), besides altering the body weight gain. Hydrocortisone treatment doubled the number of apoptotic cells after 2 hr, and quadruplicated it after 3 hr. The results demonstrated that glucocorticoids inhibit cell proliferation in the gastric epithelium of fetuses and suckling rats and increase apoptotic rates, suggesting the exit from cell cycle. PMID- 11066033 TI - Optimization of perfusion decalcification for bones and joints in rats. AB - Decalcification of osseous tissues by perfusion of decalcifying solution into the vascular system has never been applied to the study of peripheral joints. To optimize perfusion methods, rats were decalcified by direct immersion or by one of two perfusion techniques: 1) systemic perfusion circulating the decalcifying solution from the ascending aorta; and 2) regional perfusion circulating the solution to the lower extremities from the abdominal aorta. The process of decalcification was monitored by serial radiographic examinations. After decalcification, bone and joint samples were stained for histochemistry and immunohistochemistry. With systemic perfusion, the decalcification time, dependent on weight, was markedly reduced compared to immersion. Regional perfusion decalcification was faster than all other methods studied. Microstructural preservation was comparable and immunostaining quality often improved. Applications of this work will improve the study of basic skeletal and articular problems. PMID- 11066034 TI - Three-dimensional observations of spatial arrangement of hepatic zonation and vein system in mice and house musk shrews. AB - The three-dimensional (3D) relationship among the hepatic domains and the efferent central and afferent portal veins was investigated by macroscopy, microscopy, and computer-aided 3D reconstruction methods. To clearly distinguish the pericentral domain from the periportal, we used CCl(4)-treated mice and diabetic house musk shrews, which show typical pericentral necrosis and deposition of fat, respectively. The 3D findings obtained were verified against normal control animals using advantages of our unique observations by light and fluorescent microscopy, which made it possible to differentiate the two domains well. The pericentral domains in the mice and shrews appeared three-dimensionally as continuous branched columns, and the periportal domains exist in a sponge-like network that fills the parenchymal space among the columnar pericentral domains. The efferent central veins were concentrically surrounded by the pericentral domain, and segments of the central veins flowed into large sublobular and lobar veins. The walls of these large veins faced the pericentral domain at the confluence with the central veins; the remaining portions of the walls faced the periportal domain. The afferent portal veins were placed at the two-dimensional center of the network of the periportal domain and gave off smaller portal branches radially at the intersections of the network. Three types of liver lobules-classic, portal, and acinar-have been discussed repeatedly at the (2D) level. At the 3D level, it is reasonable to consider that the liver parenchyma consists of the two continuous domains corresponding to the distribution of the vessels that we found. PMID- 11066035 TI - Trigonocephaly in rabbits with familial interfrontal suture synostosis: the multiple effects of premature single-suture fusion. AB - Previous studies from our laboratory have characterized the craniofacial morphology and growth patterns of an inbred strain of rabbits with autosomal dominant coronal suture synostosis. A number of rabbit perinates from this colony have been collected sporadically over a 5-year period with premature interfrontal suture synostosis. The present study describes the very early onset of craniofacial dysmorphology of these rabbits and compares them to similar-aged normal control rabbits. A total of 40 perinatal New Zealand White rabbits were used in the present study. Twenty-one comprised the sample with interfrontal suture synostosis and ranged in age from 27 to 38 days postconception (term = 31 days) with a mean age of 33.53 days (+/-2.84 days). Nineteen rabbits served as age-matched, normal controls (mean age = 33.05 days +/-2.79 days). Lateral and dorsoventral radiographs were collected from each rabbit. The radiographs were traced, computer digitized, and 12 craniofacial measurements, angles, and indices were obtained. Mean measures were compared using an unpaired Student's t-test. All synostosed rabbits were stillborn or died shortly after birth. Grossly, these rabbits exhibited extreme frontal bossing, trigonocephaly with sagittal keeling, and midfacial shortening. No somatic anomalies were noted. Radiographically, rabbits with interfrontal suture synostosis had significantly (P < 0.05) narrower bifrontal widths, shorter cranial vault lengths, kyphotic cranial base angles, and different cranial vault indices (shapes) compared to controls. Results reveal severe and early pathological and compensatory cranial vault changes associated with premature interfrontal suture synostosis in this rabbit model. The 100% mortality rate noted in this condition may be related to the inheritance of a lethal genetic mutation or to neural compression from reduced intracranial volume. Results are discussed in light of current pathogenic hypotheses for human infants with premature metopic suture synostosis. PMID- 11066036 TI - Chondrogenesis of the branchial skeleton in embryonic sea lamprey, Petromyzon marinus. AB - This study provides concise temporal and spatial characteristics of branchial chondrogenesis in embryonic sea lamprey, Petromyzon marinus, using high resolution light microscopy, transmission electron, and immunoelectron microscopy. Prechondrogenic condensations representing the first branchial arch appeared first in the mid-region of the third pharyngeal arch at 13 days post fertilization (pf). Cartilage differentiation, defined by the presence of the unique, fibrillar, non-collagenous matrix protein characteristic of branchial cartilage, was first observed at 14 days pf. Development of lamprey branchial cartilage appeared unusual compared to that in jawed fishes, in that precartilage condensations appear as a one-cell wide orderly stack of flattened cells that extend by the addition of one dorsal and one ventral condensation. Development of lamprey gill arches from three condensations that fuse to form a single skeletal element differs from the developing gill arches of jawed fishes, where more than one skeletal element forms from a single condensation. The initial orderly arrangement of cells in the lamprey branchial prechondrogenic condensations remains throughout development. Once chondrification of the condensations begins, the branchial arches start to grow. Initially, growth occurs as a result of matrix secretion and cell migration. Later in development, the arches grow mainly by cell proliferation and enlargement. This study defines the morphology and timing of lamprey branchial chondrogenesis. Studies of lamprey chondrogenesis provide not only insight into the developmental biology of a unique non collagenous cartilage in a primitive vertebrate but also into the general evolution of the skeletal system in vertebrates. PMID- 11066037 TI - Study of the functional anatomy of bovine oviductal mucosa. AB - The oviducts of 31 cyclic cows were examined to study the structure and nature of the oviductal mucosa. The general distribution of spermatozoa within the oviductal mucosa was studied in five additional cows. The oviductal infundibulum is an asymmetric funnel-shaped structure surrounding the ostium. It is divided along the free boarder of the mesosalpinx and presents one wide and one narrow side. The mucosa of the wide side possesses a system of low interconnected cords that converge distally forming primary folds. The folds on the narrow side start sharply from the free margin and fuse toward the ostium abdominale. Areas between folds throughout the lumen of the oviduct show a high degree of complex organization. Interfold spaces are occupied by secondary and small interconnected folds which join to form a system of cul-de-sacs. In the infundibulum, these cul de-sacs open toward the ovary, while cul-de-sacs present in the caudal isthmus and in the UTJ open toward the uterus. Marked variations were observed in the oviductal epithelium depending on the oviductal segment, basal or apical areas of the folds, and phase of the oestrous cycle. Near to the time of ovulation, numerous spermatozoa were found in the periphery of the caudal isthmus within pockets of basal interfold areas, as well as within pockets and cul-de-sacs of the tubo-uterine junction. Individual spermatozoa were also observed in peripheral areas of the ampullary-isthmic junction and ampulla. The topography of the oviduct provides a complex system of regulation which may influence not only the passage of gametes and/or embryos, but also movement of fluid within the oviductal canal. PMID- 11066038 TI - Conotruncal anomalies in the trisomy 16 mouse: an immunohistochemical analysis with emphasis on the involvement of the neural crest. AB - The trisomy 16 (Ts16) mouse is generally considered a model for human Down's syndrome (trisomy 21). However, many of the cardiac defects in the Ts16 mouse do not reflect the heart malformations seen in patients suffering from this chromosomal disorder. In this study we describe the conotruncal malformations in mice with trisomy 16. The development of the outflow tract was immunohistochemically studied in serially sectioned hearts from 34 normal and 26 Ts16 mouse embryos ranging from 8.5 to 14.5 embryonic days. Conotruncal malformations observed in the Ts 16 embryos included double outlet right ventricle, persistent truncus arteriosus, Tetralogy of Fallot, and right-sided aortic arch. This spectrum of malformations is remarkably similar to that seen in humans suffering from DiGeorge syndrome (DGS). As perturbation of neural crest development has been proposed in the pathogenesis of DGS we specifically focussed on the fate of neural crest derived cells during outflow tract development of the Ts16 mouse using an antibody that enabled us to trace these cells during development. Severe perturbation of the neural crest-derived cell population was observed in each trisomic specimen. The abnormalities pertained to: 1) the size of the columns of neural crest-derived cells (or prongs); 2) the spatial orientation of these prongs within the mesenchymal tissues of the outflow tract; and 3) the location in which the neural crest cells interact with the myocardium. The latter abnormality appeared to be responsible for ectopic myocardialization found in trisomic embryos. Our observations strongly suggest that abnormal neural crest cell behavior is involved in the pathogenesis of the conotruncal malformations in the Ts16 mouse. PMID- 11066039 TI - Nerve fiber composition of the intracranial portion of the oculomotor, trochlear, and abducens nerves in the sheep. AB - In the present investigation, the fiber content and the diameter spectra of the intracranial portion of the three oculomotor nerves (oculomotor, trochlear, and abducens nerves) were analysed in sheep by light and electron microscopy. It was determined that up to 14.98% of fibers in the oculomotor nerve, 17.01% in the trochlear nerve, and 11.87% in the abducens nerve were unmyelinated. The myelinated fibers showed a bimodal distribution in their size spectrum in all three nerves, with a majority of large myelinated axons, but a considerable proportion of small myelinated fibers, as well. The sensory function of the unmyelinated fibers present in the three oculomotor nerves is discussed also on the basis of our previous morphofunctional investigations. PMID- 11066040 TI - Ultrastructure of the capillary pericytes and the expression of smooth muscle alpha-actin and desmin in the snake infrared sensory organs. AB - The infrared sensory membranes of pit organs of pit vipers have an extremely rich capillary vasculature that forms many vascular loops, each serving a small number of infrared nerve terminals. We clarified the ultrastructure of capillary pericytes in the pit membranes by scanning and transmission electron microscopy, and examined the immunoreactivity in their cytoplasm to two contractile proteins: smooth muscle alpha-actin (SM alpha-actin) and desmin. The capillary pericytes had two major cytoplasmic processes: thickened primary processes that radiate to embrace the endothelial tube and flattened secondary processes that are distributed widely on the endothelium. Coexpression of SM alpha-actin and desmin was observed in the pericytes of entire capillary segments, and SM alpha-actin was characterized by prominent filament bundles directed mainly at right angles to the capillary long axis. This expression pattern was different from that of capillary pericytes of the scales, where SM alpha-actin was expressed diffusely in the cytoplasm. In a series of electron microscopic sections, we often observed the pericyte processes depressing the endothelial wall. We also observed a close relationship of the pericytes with inter-endothelial cell junctions, and pericyte processes connected with the endothelial cells via gap junctions. From these findings, we surmised that capillary pericytes in the pit membrane have a close functional relationship with the endothelium, and through their contractile and relaxing activity regulate capillary bloodflow to stabilize production of infrared nerve impulses. PMID- 11066041 TI - Neuromuscular specializations of the pharyngeal dilator muscles: II. Compartmentalization of the canine genioglossus muscle. AB - The genioglossus (GG) muscle is divided into horizontal and oblique compartments that are the main protrusor and depressor muscles of the tongue, respectively. In humans the GG plays an important role in speech articulation, swallowing, and inspiratory dilation of the pharynx. At present, little is known about the neuromuscular specializations of the GG in any mammal. This study examined the specializations of these compartments in the canine tongue using a variety of anatomical and histochemical techniques. Six canine GG muscles were sectioned and stained for myofibrillar ATPase to study muscle fiber types; five whole-mount GG muscles were stained for acetylcholinesterase (AChE) to study the distribution of motor endplates; and eight whole mount GG muscles were processed with Sihler's stain to study the entire nerve supply pattern. In addition, the arrangement of muscle fibers of the GG within the tongue was also determined (N = 3). The most notable difference between the compartments of the GG was their proportions of fast and slow twitch muscle fibers: the horizontal compartment contained 64% slow twitch muscle fibers compared to 41% in the oblique compartment. In addition, although the oblique compartment appeared to be grossly homogeneous, it could be divided into thirds by significant differences in the percentages of slow twitch fibers: posterior (23%), middle (15%), and anterior (56%; P < 0.05). The muscle fibers of the oblique GG within the tongue were found to be divided into medial and lateral layers that run vertically and transversely, respectively. The nerve supply to each third of the oblique GG formed a plexus with the anterior third being the densest. The innervation pattern of the oblique GG was also notable as terminal nerve branches coursing parallel to the muscle fascicles gave off perpendicular secondary branches along each motor endplate band. These secondary nerve branches connected the primary nerves and formed a regularly spaced grid throughout the compartment. Evidently, the two compartments of the GG exhibited different anatomical specializations. The horizontal had a slow muscle fiber profile and simple innervation pattern; these qualities are possibly related to its single force vector and respiratory related activity. The oblique compartment had a relatively fast muscle fiber profile with evidence for three separate functional subdivisions. The most anterior part was noticeably different, and was presumably specialized for fine motor control of the tip of the tongue. The vertically oriented fibers of the oblique GG within the tongue body may function as a midline depressor of the tongue, whereas its transversely oriented fibers could play a role in narrowing the tongue during other motor tasks. PMID- 11066042 TI - Sporadic colorectal adenocarcinomas with high-frequency microsatellite instability. AB - BACKGROUND: Widespread microsatellite instability (MSI) occurs in nearly 15% of sporadic colorectal cancers. Large bowel carcinomas with high-frequency MSI (MSI H) (instability at > or = 30% of microsatellite loci) are believed to display distinctive pathologic features and to behave less aggressively than microsatellite-stable (MSS) tumors and carcinomas with low-frequency MSI (MSI-L) (instability at < 30% of microsatellite loci). The aim of the current study was to accurately define the clinicopathologic and biologic features of MSI-H sporadic colorectal carcinomas. METHODS: MSI status was evaluated in 216 large bowel adenocarcinomas using polymerase chain reaction (PCR) and 6 microsatellite markers. Tumors that showed instability with at least two microsatellite markers were classified as MSI-H, whereas the other tumors were classified as MSI-L (instability at one locus) or MSS (no instability). Expression of p53, hMLH1, and hMSH2 gene products was determined by immunohistochemistry, and DNA ploidy pattern was determined by flow cytometry. The prognostic significance of MSI status was assessed by univariate and multivariate survival analyses. RESULTS: The significantly different pathologic features of MSI-H carcinomas were proximal location; large size; mucinous and medullary histotype; poor differentiation; expanding pattern of growth; more frequent Crohn-like conspicuous lymphoid reaction; and low incidence of extramural vein invasion. Most MSI-H tumors were DNA diploid (33 of 40 tumors; 82.5%) and p53 negative (34 of 44 tumors; 77.3%). Conversely, DNA aneuploidy and p53 overexpression were observed in 82.3% (130 of 158 tumors; P < 0.0001) and 54.1% (93 of 172 tumors; P = 0.0002) of MSI-L/MSS tumors, respectively. Loss of hMLH1 or hMSH2 expression was detected in a high fraction of MSI-H carcinomas (86. 0%). Patients with MSI-H tumors showed a better clinical outcome than patients with MSI-L/MSS tumors (P = 0.0017). Furthermore, in multivariate analysis that included conventional clinicopathologic parameters, MSI status, and p53 expression as covariates, MSI status was a significant independent prognostic indicator of disease specific survival. CONCLUSIONS: Assessment of MSI status is an essential step in the genetic characterization of large bowel carcinomas and identifies a subset of tumors with distinct clinical, pathologic, and biologic features. PMID- 11066043 TI - Host defense and survival in patients with lung carcinoma. AB - BACKGROUND: Numerous studies have investigated locoregional immune responses and long term survival in patients with various types of cancer; few have focused on patients with lung carcinoma. The current study was designed to assess the prognostic value of immunomorphologic changes in locoregional lymph nodes and lymphocytic infiltration of primary tumor (LI) in patients who undergo resection for bronchogenic carcinoma. METHODS: In a retrospective analysis, immune responses in locoregional lymph nodes and at primary tumor sites were studied histologically in 172 selected patients. Lymph node morphology was studied according to the system of Cottier et al. Sinus histiocytosis and paracortical lymphoid cell hyperplasia were considered to be cellular immune responses, and follicular hyperplasia of the cortical area was considered to be a humoral reaction. LI was classified with Black's method. The survival rate was estimated by using the Kaplan-Meier product-limit method. The log rank test and the Cox proportional-hazards model were used to determine statistical significance in univariate and multivariate survival analyses. RESULTS: Among the 172 patients, 35.5% had no evident response in regional lymph nodes, 19.8% had a marked cellular response, 11% had a marked humoral response, and 33.7% had a mixed cellular and humoral response. LI was intense in 36.6% of patients and was absent or scarcely evident in 63.4%. A lymph node cellular response and marked LI improved long term survival rates even in patients with regional lymph node metastases. Multivariate analysis identified two independent variables that had high prognostic value: lymph node immunoreactivity and LI. CONCLUSIONS: Lymph node immunoreactivity and LI significantly influence long term survival after curative surgery for patients with carcinoma of the lung and may be useful in stratifying patients for prospective trials of adjuvant treatment, including immunotherapy. PMID- 11066044 TI - Biopsy confirmed benign breast disease, postmenopausal use of exogenous female hormones, and breast carcinoma risk. AB - BACKGROUND: A history of proliferative benign breast disease has been shown to increase the risk of developing breast carcinoma, but, to the authors' knowledge, how postmenopausal exogenous female hormone use, in general, has affected breast carcinoma risk among women with a history of proliferative breast disease with or without atypia has not been well established. METHODS: In the current case control study, nested within the Nurses' Health Study, benign breast biopsy slides of 133 postmenopausal breast carcinoma cases and 610 controls with a history of benign breast disease, were reviewed. Reviewers had no knowledge of case status. RESULTS: Women with proliferative disease without atypia had a relative risk for postmenopausal breast carcinoma of 1.8 (95%, confidence interval [CI]: 1.1 to 2.8), and women with atypical hyperplasia had a relative risk of 3.6 (95%, CI: 2.0 to 6.4) compared with women who had nonproliferative benign histology. Neither current postmenopausal use of exogenous female hormones nor long term use for 5 or more years further increased the risk of breast carcinoma in the study population beyond that already associated with their benign histology. CONCLUSIONS: Women who had proliferative benign breast disease, with or without atypia, were at moderately to substantially increased risk of developing postmenopausal breast carcinoma compared with women who had nonproliferative benign conditions. In the current study, postmenopausal exogenous female hormone use in general did not further increase the breast carcinoma risk for women with proliferative benign breast disease. However, the analysis did not exclude the possibility of increased risk with a particular hormone combination or dosage. PMID- 11066045 TI - Cadherin expression in glandular tumors of the cervix. AB - BACKGROUND: The cadherins are homotypic adhesion proteins that are important in cell sorting during organogenesis. Classic cadherins include several different types that show tissue specific expression. Specific tissue expression of cadherins often is preserved in neoplastic transformation, and cadherin phenotype can be used to differentiate morphologically similar but histogenetically distinct tumors. METHODS: The authors examined by using immunohistochemistry in paraffin sections the expression of E- (epithelial) and P- (placental) cadherin in 39 patients with glandular tumors of the cervix, including invasive adenocarcinoma, villoglandular adenocarcinoma, adenocarcinoma in situ (AIS), and adenoma malignum. RESULTS: In all cases, E-cadherin was expressed in both normal and malignant glands without appreciable differences. P-cadherin, normally confined to basal epithelial cells and not observed in benign glands, was aberrantly expressed in neoplastic glands in 27 cases, including 96%(23 of 24 cases) of invasive cancers, 40% (2 of 5) of villoglandular carcinomas, 25% (2 of 8) of AIS, and 0% (0 of 2) of adenoma malignum. CONCLUSIONS: The authors' results show that E-cadherin is uniformly expressed in glandular tumors of the cervix with no evidence of decreased expression in these tumors. In addition, P-cadherin is aberrantly expressed in most adenocarcinomas and appears to be preferentially expressed in invasive rather than in situ lesions. Thus, aberrant expression of P cadherin may be a useful marker of invasive or aggressive clinical behavior in glandular lesions of the cervix. PMID- 11066046 TI - Expression of c-Ets1 is associated with malignant potential in endometrial carcinoma. AB - BACKGROUND: The protooncogene c-ets1 is a transcriptional factor that controls the expression of a number of genes involved in extracellular matrix remodeling. It might play a role in the regulation of physiologic processes such as cell proliferation and differentiation and also is associated with angiogenesis, cell migration, and tumor invasion. METHODS: To elucidate the involvement of c-Ets1 in endometrial carcinogenesis, the authors analyzed serial frozen sections for c Ets1 protein expression in 20 cases of endometrial carcinoma and 20 cases of normal endometria by fluorescent immunohistochemistry. The authors analyzed the relation between the percentages of c-Ets1 stained cells and patient characteristics including histologic grade, surgical stage, presence of invasion to greater than one-half myometrium, presence of vascular involvement, presence of lymph node metastasis, and clinical outcome. RESULTS: In the normal endometria, c-Ets1 was weakly detected at the luminal surface of endometrial glands in both the proliferative and secretory phases. Most of the c-Ets1 proteins were found in the cytoplasm and partly in the nucleus of endometrial carcinoma glands, and also in fluid secreted from endometrial carcinoma glands. Moreover, c-Ets1 was strongly expressed in the head portion of papillary carcinoma tissues that invaded the stroma. c-Ets1 expression was associated significantly with histologic grade (P < 0.005), the presence of invasion to greater than one-half myometrium (P < 0.001), surgical stage (P < 0.005), and vascular involvement (P = 0.009). CONCLUSIONS: The authors' results show that c Ets1 expression in endometrial carcinoma correlates with the malignant potential of this tumor. PMID- 11066047 TI - Ovarian carcinoma diagnosis. AB - BACKGROUND: Ovarian carcinoma often is called the "silent killer" because the disease usually is not detected until an advanced stage. The authors' goal was to evaluate preoperative symptoms and factors that may contribute to delayed diagnosis for women with ovarian carcinoma. METHODS: A two-page survey was distributed to 1500 women who subscribe to CONVERSATIONS!, a newsletter about ovarian carcinoma. Because the survey could be copied and given to other patients, 1725 surveys were returned from women in 46 states and 4 Canadian provinces. RESULTS: The median age of the surveyed women was 52 years, and 70% had Stage III or IV disease (International Federation of Gynecology and Obstetrics). When asked about symptoms before the diagnosis of ovarian carcinoma, 95% reported symptoms, which were categorized as abdominal (77%), gastrointestinal (70%), pain (58%), constitutional (50%), urinary (34%), and pelvic (26%). Only 11% of women with Stage I/II and 3% with Stage III/IV reported no symptoms before their diagnosis. Women who ignored their symptoms were significantly more likely to be diagnosed with advanced disease compared with those who did not (P = 0.002). The time required for a health care provider to make the diagnosis was reported as less than 3 months by 55%, but greater than 6 months by 26% and greater than 1 year by 11%. Factors significantly associated with delay in diagnosis were omission of a pelvic exam at first visit; having a multitude of symptoms; being diagnosed initially with no problem, depression, stress, irritable bowel, or gastritis; not initially receiving an ultrasound, computed tomography, or CA 125 test; and younger age. The type of health care provider seen initially, insurance, and specific symptoms did not correlate with delayed diagnosis. CONCLUSIONS: This large national survey confirms that the majority of women with ovarian carcinoma are symptomatic and frequently have delays in diagnosis. PMID- 11066048 TI - Carcinoma of the fallopian tube. AB - BACKGROUND: The objective of the current study was to increase insight into the biology of fallopian tube carcinoma through an analysis of possible clinical and pathologic determinants of prognosis and to formulate recommendations with regard to a more optimal therapeutic approach for patients with this rare disease. METHODS: A study was performed of the pathology specimens and clinical case records from 151 patients with fallopian tube carcinoma who were treated consecutively. Both univariate and multivariate analyses of possible prognostic factors were performed for the whole group and for the subgroup of 41 patients with Stage I disease. The possible significance of serum CA-125 levels as a tumor marker and a marker of response to platinum-containing chemotherapy was evaluated. RESULTS: In multivariate analysis, disease stage, the presence of residual tumor, and a hydrosalpinx-like appearance of the fallopian tube were of independent prognostic significance for the whole cohort. For patients with Stage I disease, the depth of infiltration in the tubal wall and intraoperative tumor rupture were of independent prognostic significance. The marked tendency of this disease for extraperitoneal spread, even in apparently early stages, was confirmed. In 37 evaluable, platinum-naive patients, an overall response rate of 70% was obtained with platinum-based chemotherapy, with a median response duration of 12.5 months. In view of its low efficacy and high rate of serious complications, the use of postoperative radiotherapy in the treatment of patients with fallopian tube carcinoma is no longer recommended. Serum CA-125 level measurements in fallopian tube carcinoma patients have the same significance as tumor and surrogate markers of response as in ovarian carcinoma patients. CONCLUSIONS: Prognostic factors in patients with early stage (Stages 0 and I) fallopian tube carcinoma seem to differ from those in patients with early stage ovarian carcinoma. For patients with more advanced stage disease, due to the striking similarities in prognostic and clinical characteristics between the two diseases, the authors recommend that the treatment and follow-up strategies for patients with ovarian carcinoma be adopted in the management of patients with fallopian tube carcinoma. PMID- 11066049 TI - Prostatourethral-rectal fistula after prostate brachytherapy. AB - BACKGROUND: Brachytherapy (BT) has seen increased utilization as a potentially curative treatment for patients with localized initial or recurrent prostate carcinoma. This modality can be delivered by palladium 103 (Pd(103)) or iodine 125 (I(125)) implant with or without external beam radiotherapy (EBRT). Prostatourethral-rectal fistula (PRF) is a serious complication of this approach, and its incidence, clinical presentation, and risk factors for occurrence have not been documented thoroughly. Thus, the authors sought to determine these factors in a large series of patients who were treated at two institutions. METHODS: Seven hundred sixty-five patients received outpatient BT using a computed tomography (CT)-guided or transrectal ultrasound (TRUS)-guided technique between July 1994 and June 1999 using either Pd(103) or I(125) implants. Of the 754 patients with follow-up, 640 patients received BT monotherapy, 69 patients received BT monotherapy as a boost after EBRT, and 45 patients received BT as salvage therapy after locally recurrent prostate carcinoma that was treated initially with BT (20 patients), EBRT (20 patients), surgery plus EBRT (3 patients), surgery and high dose rate radiotherapy (HDR) (1 patient), or EBRT plus HDR (1 patient). CT dosimetry of the TRUS-guided implants was carried out in all patients 1-7 days postprocedure. Patient follow-up and clinical status were compiled in a data base. RESULTS: Seven PRFs developed in 754 patients (1%) between 9 months and 12 months after treatment. One PRF (0.2%) occurred in patients who were treated with BT monotherapy. PRFs occurred in patients who were treated with combination therapy (2 of 69 patients; 2.9%) and in patients who underwent salvage BT (4 of 45 patients; 8.8%) patients. All six patients who developed fistulas in the context of combination BT/EBRT or salvage BT had biopsy of an anterior rectal lesion overlying the prostate noted on physical examination during routine follow-up. Gastrointestinal endoscopic evaluation alone was not associated with any PRF. Five of the seven PRFs resolved with either surgical repair (3 patients) or conservative management (2 patients). CONCLUSIONS: There is a low incidence of PRF formation after BT monotherapy. Because all patients who developed PRF did so subsequent to prior rectal biopsies, the authors currently are discouraging such practices strongly if the rectal lesion is consistent with radiation-induced effects. PMID- 11066050 TI - Implication of telomerase activity and alternations of telomere length in the histologic characteristics of intracranial meningiomas. AB - BACKGROUND: Telomerase activity and telomere length have been shown to be involved in the control of cell proliferation and regulation of cell senescence. The expression of telomerase activity may endow cells with the capacity of unlimited proliferation and immortality. The authors examined the telomerase activity and telomere length of intracranial meningiomas to determine the relation between the results and the clinicopathologic behavior of these tumors. METHODS: Sixty-two specimens of meningiomas including 13 atypical and malignant tumors were used in this study. Telomerase activity was measured with polymerase chain reaction and enzyme-linked immunosolvent assay. Telomere length was measured by detecting the terminal restriction fragments using Southern blots. RESULTS: Detectable telomerase activity was found in 4 of 13 (30.8%) malignant or atypical meningiomas and only 1 in 49 benign meningiomas (P = 0.006). Elongated telomere length was measured in 6 of 13 (46.1%) patients with malignant or atypical meningiomas and only 1 of 48 (2.1%) in those with benign tumors (P = 0.0002). Three of 4 (75%) of malignant or atypical meningiomas with detectable telomerase activity revealed shortened telomere length, and all tumors with elongated telomere length displayed undetectable telomerase activity. The percentage of malignant or atypical meningiomas with detectable telomerase activity or elongated telomere were significantly higher (76.9%) than that of benign tumors (4.0%). The proliferative index was calculated as the percentage of tumor cell nuclei immunoreactive for Ki-67 to total tumor nuclei. The mean values of proliferative index in benign, atypical, and malignant meningiomas were 1.2, 11.0, and 30.0, respectively. CONCLUSIONS: The results indicate that telomerase activation may be a critical step in the pathogenesis of malignant or atypical meningioma. Elongation of the telomere length also implicates the high potential for malignant behavior in these tumors. PMID- 11066051 TI - A combination therapy of continuous superselective intraarterial carboplatin infusion and radiation therapy for locally advanced head and neck carcinoma. Phase I study. AB - BACKGROUND: To improve the treatment result for locally advanced head and neck carcinoma, the authors used a combination of radiotherapy with superselective continuous intraarterial therapy using carboplatin. The dose limiting toxicity (DLT), maximum tolerated dose (MTD), and treatment effectiveness were tested in Phase I and II protocols. PATIENTS AND METHODS. Thirty-five patients were entered into the study from August 1992 to May 1997. The target arteries were the lingual artery in 18 cases, facial artery in 5 cases, maxillary artery in 11 cases, and external carotid artery initially changing to lingual artery in 1 case. Escalating daily carboplatin doses were tested, starting from 10 mg/m(2) (total dose, 360 or 400 mg/m(2)) to 15 mg/m(2) (total dose, 405 or 450 mg/m(2)) and then 20 mg/m(2) (total dose, 460 or 500 mg/m(2)). Radiotherapy was administered using a 6-megavolt linear accelerator to a total dose of 50-60 grays. Interstitial radiotherapy boost also was used for carcinoma of the tongue. RESULTS: Excluding 3 patients who discontinued treatment, the treatment results of 32 patients were complete response in 21 cases, partial response in 10 cases, and no change in 1 case. Neutropenia was the DLT, and the MTD was 500 mg/m(2). The local control rate was 64%. CONCLUSIONS: Superselective continuous intraarterial carboplatin and concurrent radiation therapy can be delivered safely with good efficacy for locally advanced carcinomas of the tongue and base of the tongue. Surgical treatment of these diseases usually incurs severe functional loss. This current approach may be a breakthrough in these cancers. PMID- 11066052 TI - Clinicopathologic and genotypic study of extranodal nasal-type natural killer/T cell lymphoma and natural killer precursor lymphoma among Koreans. AB - BACKGROUND: This study aimed to define genotypic profile and to describe the clinicopathologic features of nasal-type natural killer (NK)/T-cell lymphoma of nasal and extranasal origin and NK precursor lymphoma. METHODS: NK/T-cell lymphomas from the upper aerodigestive tract (n = 45), skin (n = 2), gastrointestinal tract (n = 3), and soft tissue (n = 2) and NK precursor neoplasms (n = 3) were studied. Immunophenotype was analyzed by immunohistochemistry and flow cytometry. In situ hybridization with EBER 1/2 RNA probes was performed. T-Cell Receptor (TCR)-gamma gene rearrangement was analyzed by seminested polymerase chain reaction with heteroduplex analysis. Overall survival rate was correlated with clinicopathologic parameters and compared by Wilcoxon test. RESULTS: Clonal TCR-gamma gene rearrangement was detected in 3 of 31 upper aerodigestive and 1 of 2 skin tumors. When immunostained using paraffin embedded tissue, 6 upper aerodigestive lymphomas were negative for CD56 in which 4 cases lacked clonal TCR gene rearrangement. Epstein-Barr virus (EBV) mRNA was detected in 33 upper aerodigestive tumors including 26 of 29 nasal tumors (90%), and 7 of 10 extranasal tumors (70%). There was no histologic, immunophenotypic, or genotypic differences according to the lineage and EBV association in upper aerodigestive lymphomas. Among the patients with upper aerodigestive tumors, overall 1-year survival rate was 41%, and correlated well with the stage (P < 0.05) but not with the size of tumor cells, EBV status, and lineage (P > 0.05). Median survival rate of lymphomas from other sites excluding upper aerodigestive tract was not significantly different from that of upper aerodigestive lymphomas with same stage (P > 0.05). Unlike nasal-type NK/T-cell lymphomas, NK precursor lymphoma involved the bone marrow and lymph nodes at initial presentation or in the course of disease. Tumor cells were positive for TdT in all and myeloid markers in two. TCR gene rearrangement was germ line. CONCLUSIONS: Most upper aerodigestive nasal-type NK/T-cell lymphomas among Koreans are genotypically of NK derivation and few belong to T lineage. Presence or absence of EBV has no significant correlation with the histologic changes and the lineage of these lymphomas. PMID- 11066053 TI - Successful treatment of metastatic retinoblastoma. AB - BACKGROUND: In the past, patients with metastatic retinoblastoma have had a poor prognosis when treated with conventional modalities. In the current study, the authors evaluated the use of combined intensive conventional chemotherapy, high dose chemotherapy with autologous stem cell rescue (ASCR), and radiation therapy. METHODS: Four patients with metastatic retinoblastoma were treated. All had orbital and bone marrow metastases. In addition, three patients had bone metastases and two patients had liver metastases. None had central nervous system disease. Patients received intensive conventional chemotherapy that included vincristine, cyclophosphamide, etoposide, and either cisplatin or carboplatin. Stem cells were harvested after bone marrow disease was no longer detectable. High dose chemotherapy with carboplatin (500 mg/m(2)/day x 3 days or area under the curve = 7 via the Calvert formula) and thiotepa (300 mg/m(2)/day x 3 days) with (n = 3 patients) or without (n = 1 patient) etoposide (250 mg/m(2)/day x 3 days) was administered with ASCR. Sites that originally harbored bulky disease were irradiated after recovery from the high dose chemotherapy. RESULTS: The therapy was associated with substantial acute hematopoietic and mucosal toxicities. At last follow-up, all four patients had survived event free from 46 80 months after the diagnosis of metastatic disease. CONCLUSIONS: The treatment strategy described in the current study is effective for patients with metastatic retinoblastoma that does not involve the central nervous system. However, a multicenter trial should be considered to evaluate it in a larger group of patients. PMID- 11066054 TI - Granulocyte-colony stimulating factor and multiple cycles of strongly myelosuppressive alkylator-based combination chemotherapy in children with neuroblastoma. AB - BACKGROUND: The authors assessed key effects of granulocyte-colony stimulating factor (G-CSF) used prophylactically with multiple cycles of strongly myelosuppressive alkylator-based combination chemotherapy. To the authors' knowledge, no large study has focused on G-CSF in this setting, yet this kind of treatment has recently become standard for poor risk pediatric solid tumors such as neuroblastoma. PATIENTS AND METHODS. Children with neuroblastoma received cyclophosphamide 140 mg/kg (i.e., 4200 mg/m(2)), doxorubicin 75 mg/m(2), and vincristine (CAV) in cycles 1, 2, 4, and 6 and cisplatin 200 mg/m(2) and etoposide 600 mg/m(2) (P/VP) in cycles 3, 5, and 7. To maximize dose intensity, chemotherapy was begun as soon as the absolute neutrophil count (ANC) was > or = 500/microL and platelet count was > or = 100,000/microL. No cytokines were used during 1990-1994 (control group; n = 28), but G-CSF was used from 1995 to 1998 (G CSF group; n = 30) at 5 microg/kg/day subcutaneously from 1 day after chemotherapy until the ANC was > or = 500/microL on 2 successive days or was > or = 1000/microL. RESULTS: Each cycle of CAV decreased ANCs to < 200/microL in all 58 patients; recovery to 200/microL and to 500/microL was significantly sooner with G-CSF. In contrast, P/VP did not invariably cause severe neutropenia: similar numbers of patients in each group maintained ANCs > or = 200/microL and > or = 500/microL; recovery to 500/microL (but not to 200/microL) was significantly faster in the G-CSF group. G-CSF had no impact on rates of febrile episodes. Bacterial/fungal infections were slightly less frequent in the G-CSF group with CAV (P = 0.11) but not with P/VP. Dose intensity through cycle 4 was the same in both groups. Beginning with cycle 3, G-CSF patients had slower recovery to platelet counts > or = 100,000/microL. Response rates were similar in the two groups. CONCLUSIONS: With multiple cycles of strongly myelosuppressive alkylator based combination chemotherapy, prophylactic use of G-CSF hastened ANC recovery but did not reduce the incidence of febrile episodes, had little impact on infection rates, did not yield augmented dose intensity, was associated with prolonged thrombocytopenia, and had no effect on response rates of neuroblastoma. The data support more limited use of G-CSF. PMID- 11066055 TI - Oral trofosfamide and etoposide in pediatric patients with glioblastoma multiforme. AB - BACKGROUND: Glioblastoma multiforme in childhood is rare, and the prognosis for patients with the disease is poor. The Pediatric Oncology Society of the Germanic language group (GPOH) enrolls patients in a series of pilot trials, the first of which is reported here (HIT-GBM-A). METHODS: Twenty-two patients with glioblastoma multiforme, World Health Organization Grade 4, between the ages of 3 15 years (45% male) were enrolled during the period 1995-1997. There were 13 supratentorial tumors, 8 brainstem tumors, and 1 cerebellar tumor. The patients underwent the following procedures: stereotactic biopsy (n = 3 patients), open biopsy (n = 1 patient), partial resection (n = 6 patients), subtotal resection (n = 4 patients), and macroscopic total resection (n = 8 patients). Adjuvant treatment consisted of oral chemotherapy with trofosfamide, 100 mg/m(2), and etoposide, 25 mg/m(2), daily or for 21-day cycles interrupted by 1-week rests. Standard fractionated radiation (54 grays) was started concurrently with the first cycle. RESULTS: The chemotherapy was well tolerated, with no treatment related deaths and only minor side effects. The responses in 12 evaluable patients after two cycles were as follows: 1 complete response, 1 partial response, 3 patients with stable disease, and 7 patients with progressive disease. The median overall survival was 12 months. The 1-year, 2-year, and 4 year overall survival rates were 52%, 26%, and 22%, respectively, and the event free survival rates were 26%, 22%, and 4%, respectively. None of the four surviving patients (3.2 years, 3.4 years, 4.0 years, and 4.2 years after diagnosis) is event free. Two patients are alive after tumor progression: One patient was diagnosed with a medulloblastoma, and one patient was diagnosed with an osteosarcoma as second malignancies. A control group extracted from the Surveillance, Epidemiology, and End Results data had lower survival rates: the difference between the groups was not statistically significant (P = 0.26). CONCLUSIONS: This chemotherapy will not be used in a randomized trial of patients with glioblastoma; however, it may be evaluated for patients with tumors that have more chemoresponsive histologies. PMID- 11066056 TI - Lymphomatoid papulosis associated with both severe hypereosinophilic syndrome and CD30 positive large T-cell lymphoma. AB - BACKGROUND: Previous reports have found an association between lymphomatoid papulosis and hypereosinophilic syndrome, as well as lymphomatoid papulosis and lymphoma. In the current study the authors report what to their knowledge is the first reported case of these three diseases occurring simultaneously in the same patient. METHODS: The authors followed the clinical course of a 64-year-old man with lymphomatoid papulosis associated with severe hypereosinophilic syndrome complicated by involvement of the lungs and heart. RESULTS: After 6 years of follow-up, the patient developped a large T-cell, CD30 positive lymphoma. The bone marrow biopsy was typical of hypereosinophilic syndrome associated with fibrosis, with focal lymphomatous infiltrates comprised of large cells resembling the type A cells of lymphomatoid papulosis. Complete remission of the lymphoma was obtained with chemotherapy. CONCLUSIONS: This exceptional case report suggests a link between the three diseases. Lymphomatoid papulosis belongs to the spectrum of CD30 positive lymphoproliferative disorders and CD30 positive lymphocytes of lymphomatoid papulosis are known to have a Th2 profile with possible secretion of eosinopoietic cytokines. PMID- 11066057 TI - Errata PMID- 11066058 TI - Fusion of membranes during the acrosome reaction: a tale of two SNAREs. AB - During spermiogenesis, hydrolytic enzymes are sorted from the Golgi apparatus to the acrosome, a supranuclear megavesicle. At fertilization, the enzymatic content of the acrosome is released by exocytosis when a portion of the plasma membrane enveloping the sperm head fuses with the outer membrane of the acrosome. Membrane fusion involves the interaction of a specific pair of proteins, called SNAREs (for soluble N-ethylmaleimide sensitive factor attachment protein receptor). v SNARE is presumably associated with the membrane of the acrosomal vesicle. Target t-SNARE is associated with the plasma membrane. The interaction of v-SNARE and t SNARE requires two additional proteins: Rab proteins, members of a family of small GTPases related to the Ras proteins, and a complex of two proteins, NSF SNAP, recruited by the interacting v-SNARE-tSNARE pair. Syntaxin 2, a v-SNARE member, and Rab3A, a member of the Rab GTPases, have been localized in the acrosome of rodent sperm. PMID- 11066059 TI - Caprine pregnancy-associated glycoproteins (PAG): their cloning, expression, and evolutionary relationship to other PAG. AB - Pregnancy-associated glycoproteins (PAG) are structurally related to aspartic proteinases and belong to an extensive, rapidly evolving family of recently duplicated genes expressed in the placentas of artiodactyl species. The aim of the present study was to clone PAG from the goat, study their temporal and cell specific expression, and determine their phylogenetic relationship to PAG from other species. RT-PCR was used to generate PAG cDNA from pooled placental RNA obtained between days 45 and 115 of pregnancy. A total of 11 cDNA, which differed by > 5% from each other, were selected for complete bidirectional sequencing from 60 clones analyzed. A group of nine (caPAG1, caPAG3-7(var), caPAG9-11), which displayed > 80% sequence identity with each other, were expressed after day 45 of pregnancy and were localized to trophoblast binucleate cells. These PAG demonstrated an unusually high ratio of nonsynonymous (amino acid changing) to synonymous nucleotide differences. CaPAG2, by contrast, was detectable only in early pregnancy (days 18 and 19) and expressed throughout trophectoderm. It was of more ancient origin than the PAG1 group, but more recent than caPAG8. The latter was expressed at all stages examined (days 18 to 115). The data confirm that many PAG genes, with different patterns of temporal and spatial expression, are transcribed in the placenta of the goat. The data also suggest that the recently duplicated PAG genes are being selected for rapid diversification of function. PMID- 11066060 TI - mRNAs encoding aquaporins are present during murine preimplantation development. AB - The present study was conducted to investigate the mechanisms underlying fluid movement across the trophectoderm during blastocyst formation by determining whether aquaporins (AQPs) are expressed during early mammalian development. AQPs belong to a family of major intrinsic membrane proteins and function as molecular water channels that allow water to flow rapidly across plasma membranes in the direction of osmotic gradients. Ten different AQPs have been identified to date. Murine preimplantation stage embryos were flushed from the oviducts and uteri of superovulated CD1 mice. Reverse transcription-polymerase chain reaction (RT-PCR) methods employing primer sets designed to amplify conserved sequences of AQPs (1 9) were applied to murine embryo cDNA samples. PCR reactions were conducted for up to 40 cycles involving denaturation of DNA hybrids at 95 degrees C, primer annealing at 52-60 degrees C and extension at 72 degrees C. PCR products were separated on 2% agarose gels and were stained with ethidium bromide. AQP PCR product identity was confirmed by sequence analysis. mRNAs encoding AQPs 1, 3, 5, 6, 7, and 9 were detected in murine embryos from the one-cell stage up to the blastocyst stage. AQP 8 mRNAs were not detected in early cleavage stages but were present in morula and blastocyst stage embryos. The results were confirmed in experimental replicates applied to separate embryo pools of each embryo stage. These results demonstrate that transcripts encoding seven AQP gene products are detectable during murine preimplantation development. These findings predict that AQPs may function as conduits for trophectoderm fluid transport during blastocyst formation. PMID- 11066061 TI - Expression of enhanced green fluorescent protein (EGFP) and neomycin resistant (Neo(R)) genes in porcine embryos following nuclear transfer with porcine fetal fibroblasts transfected by retrovirus vector. AB - In this study, we demonstrated expression of enhanced green fluorescent protein (EGFP) and neomycin resistant (Neo(R)) genes in porcine embryos following nuclear transfer from porcine fetal fibroblasts (PFFs) transduced with the EGFP and Neo(R) genes by retrovirus-mediated infection. Nuclear transfer of the nonstarved transfected PFF into enucleated oocytes was accomplished by cell to cell fusion. Out of 188 porcine eggs reconstructed by nuclear transfer, 116 (61.7%) eggs cleaved and 25 (13.3%) developed to morula and blastocyst stages. Of these 25 morulae and blastocysts, 25 (100%) embryos emitted green fluorescence. Expression of the both EGFP and Neo(R) genes was detected as early as the 2-cell stage. As determined by EGFP gene expression, mosaicism was not observed in any embryo. These results suggest that porcine oocytes reconstructed by nuclear transfer with transfected PFFs can successfully develop to the blastocyst stage. In addition, this approach might be applicable to the production of transgenic pigs with complex genetic modifications. PMID- 11066062 TI - Electroporation of bovine spermatozoa to carry DNA containing highly repetitive sequences into oocytes and detection of homologous recombination events. AB - There are several methods of modifying bovine genomes. Pronuclear microinjection is more widely used but it is still to be improved. Searches for alternatives have lead to the development of new methods including SMGT (Sperm Mediated Gene Transfer), in which live spermatozoa are used as vehicles for DNA delivery during in vitro fertilization. In previous studies, we presented evidence that a highly repetitive Alu-like repeat favours transgenesis by homologous recombination (HR). Up to 60% integration via HR was obtained following pronuclear microinjection of a Pst1 beta-actin GFP DNA construction. In the present study, we show that HR mediated integration is also possible using SMGT, since bovine spermatozoa electroporated with the same DNA construct are able to transfer it to a high proportion of embryos obtained by in vitro fertilization. Swim-up selected bovine spermatozoa were mixed with the Pst1 beta-actin GFP construct (6 x 10(6) spermatozoids were incubated with 600 ng of muDNA), submitted or not to electroporation (300 V, 25 F) and treated or not with DNase I. The process of electroporation itself did not affect in vitro embryonic development. However, oocytes fertilized with electroporated DNA-treated spermatozoa developed beyond the 16-cell stage in proportions that were significantly lower (27% with Pst1 beta-actin GFP and 34% with beta-actin GFP) compared to the control without DNA (44%). On the other side, the use of electroporation significantly increased the uptake of DNA. The number of homologous recombination events detected by PCR went from 3.5% without electroporation to 46.5% after electroporation. In conclusion, our results confirm that spermatozoa electroporation combined with homologous recombination in a highly repetitive Pst1 sequence is a feasible method to obtain transgenic bovine embryos. PMID- 11066063 TI - Characterization of embryos derived from calf oocytes: kinetics of cleavage, cell allocation to inner cell mass, and trophectoderm and lipid metabolism. AB - Embryos derived from calf oocytes were compared with adult cow oocyte-derived embryos (1) by studying the kinetics of embryo development using time-lapse cinematography (2) by evaluating the ratio between inner cell mass (ICM) and trophectoderm (TE) cells in blastocysts (3) by measuring the triglyceride content of the blastocysts. The rate of calf oocyte-derived embryos reaching the blastocyst stage was reduced (26 vs. 46% for adult derived embryos). Calf oocyte derived embryos preferably arrested their development before the 9-cell stage. Those that developed into blastocysts had cleaved earlier to reach the 2-cell or 3-cell stages than embryos that arrested before the 9-cell stage. The 9-cell stage tended to appear later in calf oocyte-derived embryo that reached the blastocyst stage than in adult-derived embryos. This difference became significant at the morula stage. Accordingly, the fourth cell cycle duration was longer for calf oocyte-derived embryos. Day 8 blastocysts from both sources had similar total cell numbers (calf: 89 +/- 20; cow: 100 +/- 30) and cell distribution between TE and ICM. The triglyceride content of day 7 blastocysts was similar for both sources (64 +/- 15 vs. 65 +/- 6 ng/embryo, respectively). In conclusion, calf oocyte-derived embryos are characterized by a higher rate of developmental arrest before the 9-cell stage and by a longer lag phase preceding the major onset of embryonic genome expression. These changes might be related to insufficient "capacitation" of the calf oocyte during follicular growth. Despite these differences, modifications in the quality of the resulting blastocysts were not detected. PMID- 11066064 TI - Low oxygen tension during in vitro maturation is beneficial for supporting the subsequent development of bovine cumulus-oocyte complexes. AB - The effects of carbohydrates on meiotic maturation and ATP content of bovine oocytes under low oxygen tension (5%) were investigated. Furthermore, the developmental competence or intracellular H(2)O(2) contents of the oocytes matured under 5% or 20% O(2) was assessed. In vitro maturation of bovine cumulus oocyte complexes was performed in synthetic oviduct fluid (SOF) containing 20 amino acids and hormones (SOFaa). The proportion of the oocytes that matured to the metaphase II stage in SOFaa containing 1.5 mM glucose, 0.33 mM pyruvate, and 3.3 mM lactate under 5% O(2) was dramatically lower than that of oocytes matured under 20% O(2) (P < 0.01). Similarly, the ATP content of the oocytes that matured under 5% O(2) was much lower than that of oocytes matured under 20% O(2) (P < 0.05). Under 5% O(2) the proportion of metaphase II oocytes increased with increasing glucose concentration (0-20 mM) in SOFaa without pyruvate or lactate. In addition, the ATP content of oocytes cultured in 20 mM glucose was higher (P < 0.05) than that of oocytes cultured in 1. 5 mM glucose. Two glucose metabolites (pyruvate and lactate) and a nonmetabolizable glucose analog (2-deoxy-glucose), however, had no noticeable effects on meiotic maturation under 5% O(2). These results suggest that ATP production under 5% O(2) is not dependent on the TCA cycle. Addition of iodoacetate, a glycolytic inhibitor, to SOFaa containing 20 mM glucose significantly reduced (P < 0.01) the proportion of metaphase II and ATP content. Moreover, the proportion of the development to the blastocyst stage of oocytes matured under 5% O(2) was higher (P < 0.05) than that of oocytes matured under 20% O(2). H(2)O(2) contents of oocytes matured under 5% O(2) was lower (P < 0.05) than that of oocytes matured under 20% O(2). The results of the present study demonstrate that glucose plays important roles in supporting the completion of meiotic maturation in bovine cumulus-oocyte complexes under low oxygen tension and that low oxygen tension during in vitro maturation is beneficial for supporting the subsequent development of bovine oocytes. PMID- 11066065 TI - Expression and localization of prohormone convertase 1/3 (SPC3) in porcine ovary. AB - Tissue distribution and cellular localization of PC1/3 mRNA in porcine tissues were examined by ribonuclease protection assay and in situ hybridization. PC1/3 mRNA was detected mainly in the corpus luteum of pregnant sow and brain. Within the ovary, PC1/3 and relaxin transcripts colocalized within large luteal cells. Levels of PC1/3 transcripts in corpora lutea increased as gestation advanced, parallel with an observed increase in relaxin transcripts. A role for PC1/3 in proprotein processing in the ovary is discussed. PMID- 11066066 TI - Generation of stable cell lines by spontaneous immortalization of primary cultures of porcine granulosa cells. AB - We report the generation of stable cell lines obtained by spontaneous immortalization of primary cultures of porcine granulosa cells. Three hundred stable cell lines were obtained from three independent immortalization trials. Two of these cell lines retained the steroidogenic capabilities characteristic of granulosa cells, such as de novo synthesis of progesterone and conversion of androstenedione into estradiol-17beta. All the stable cell lines expressed the P450arom and 3betaHSD genes, confirming their granulosa origin. Moreover, the steroidogenic stable granulosa cells also expressed StAR and P450scc genes. Stable cells were developed in cultures using Medium 199 supplemented with 5% newborn calf serum (NBCS). The surviving cells overcame the senescent phase and entered a stage of continuous growth for over one hundred generations. No stable colonies were obtained from cultures grown in MEM or DMEM or media supplemented with 10% NBCS or 5 and 10% fetal calf serum (FCS). Medium 199 is a formulation richer in nutrients compared to MEM or DMEM and the cell growth capability of NBCS is lower than that of FCS, probably due to deficiency of growth factors. We speculate that spontaneous immortalization of granulosa cells may be facilitated by using a rich culture formulation supplemented with low concentrations of serum deficient in growth factors. We have validated the stable cell lines for studying the effect of hormonal steroids on granulosa cell steroidogenesis and the expression of the steroidogenic genes. Therefore, we believe that they are useful models to study the molecular mechanism involved in granulosa cell differentiation and steroidogenesis. PMID- 11066067 TI - Localization of a syntaxin isoform, syntaxin 2, to the acrosomal region of rodent spermatozoa. AB - The acrosome reaction includes a membrane fusion event that is a prerequisite for sperm penetration through the zona pellucida and subsequent fertilization. Since SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins have been shown to be key players in membrane fusion during regulated exocytosis in nerve terminals and secretory cells, and since the acrosome reaction has some features in common with regulated exocytosis, we hypothesized that SNARE proteins might also regulate acrosomal exocytosis. RT-PCR analysis demonstrated the expression of SNARE proteins, three isoforms of syntaxin 2 (2A, 2B, and 2C) and syntaxin 4A, in rat testes. Immunoblot analysis with anti syntaxin 2 antibody showed that the protein was expressed in rodent spermatozoa, and that it was associated with membrane components of spermatozoa prepared by sucrose density gradient centrifugation. Confocal laser scanning microscopy with double immunolabeling revealed that syntaxin 2 was colocalized with acrin 1, a 90 kDa acrosomal protein, over the acrosomal region of spermatozoa but was not associated with the posterior half of head or tail. Localization of syntaxin 2 over the acrosomal region was supported by the finding that it was shed from sperm heads during an acrosome reaction induced by calcium ionophore A23187 in vitro. In view of the putative role of syntaxin proteins in other membrane fusion systems, these data suggest that syntaxin 2 may be involved in regulating the acrosomal reaction in rodent spermatozoa. PMID- 11066068 TI - Fertilization promoting peptide and adenosine, acting as first messengers, regulate cAMP production and consequent protein tyrosine phosphorylation in a capacitation-dependent manner. AB - Fertilization promoting peptide (FPP) and adenosine have been shown to act as first messengers, regulating availability of the second messenger cAMP by initially stimulating cAMP production in uncapacitated spermatozoa and then inhibiting it in capacitated cells. This study investigated possible capacitation related changes in protein tyrosine phosphorylation in response to FPP and adenosine. Time-dependent changes in phosphorylation of proteins of approximately 30-140 kDa were observed in both uncapacitated and capacitated suspensions, the general level of phosphorylation being markedly greater in capacitated cells. In the presence of FPP, phosphorylation was stimulated in uncapacitated but inhibited in capacitated spermatozoa, compared with untreated control samples. Adenosine, cholera toxin, and CGS-21680, a stimulatory A(2a) adenosine receptor agonist, also stimulated phosphorylation in uncapacitated spermatozoa, while Gln FPP, a competitive inhibitor of FPP, blocked responses to FPP. In capacitated cells, FPP's inhibition of phosphorylation was abolished when cells were treated with FPP in the presence of pertussis toxin. Consistent with the capacitation dependent effects of FPP and adenosine on cAMP production, these results support the hypothesis that FPP and adenosine modulate sperm function by regulating the AC/cAMP signaling pathway and, consequently, protein tyrosine phosphorylation. Of particular significance is the identification of several phosphoproteins showing FPP-induced alterations in phosphorylation. In uncapacitated spermatozoa, proteins of approximately 116, 95, 82, 75, 66, 56, and 42 kDa showed increased phosphorylation, while in capacitated cells, phosphoproteins of approximately 116, 95, 82, 75, 70, 66, 56, and 50 kDa showed decreased phosphorylation. This suggests that these particular proteins may be involved in stimulation and arrest of capacitation, respectively. PMID- 11066069 TI - Isolation and characterisation of two sperm membrane proteins recognised by sperm associated antibodies in infertile men. AB - Antisperm antibodies eluted from the surface of spermatozoa obtained from infertile men recognised several common epitopes. We tested whether these epitopes were relevant to fertility by isolating the immunodominant 37/36 and 19/18 protein zones. These protein zones were cut out of preparative slab gels and electro-eluted. The isolated proteins, P36 and P18, were used for biochemical characterisation and to produce specific antibodies in rabbits. The specific reactivity of P36 and P18 with WGA and AAL lectins, respectively, indicated the presence of lactosaminyl structures with sialic acid termini in P36 and of fucosylated residues in P18. Isoelectric focusing showed that the two proteins consist of several polypeptides. Some of these polypeptides were recognised by both human and rabbit antibodies: the pl of these epitopes was around 5.5 for P36 and 8.3-10.3 for P18. Rabbit antibodies detected the corresponding proteins on the sperm heads of methanol-fixed and of live acrosome-reacted spermatozoa. Anti P36 antibodies bound mainly to the equatorial segment. They reduced the binding and, consequently, the penetration of zona-free hamster oocytes by human spermatozoa. Anti-P18 antibodies gave more diffuse staining of the acrosomal and post-acrosomal regions and reduced sperm-oocyte penetration without a significant effect on sperm binding. These results suggest that P36 and P18 antigens located in different compartments of the sperm head may participate in the sperm-oolemma interaction. We are currently investigating the physiological role of these antigens by sequencing the proteins isolated from the gels. PMID- 11066070 TI - Increased in vitro binding and fertilizing ability of mouse sperm exposed to a synthetic peptide. AB - We report use of an in vitro assay (Barbato et al., 1998: Biol Reprod 58:686-699) to assess binding ability of cauda epididymal mouse sperm to a surrogate zona pellucida and effect of a synthetic peptide (Amann et al., 1999: J Androl 20: 42 46) on fertilization ability in in vitro fertilization (IVF) tests. Sperm from C57Bl/6, CD1, and CF1 mice (4 replicates each) were evaluated for binding ability after exposure to 0 (control) and 80-1280 pM peptide. For control sperm, endogenous binding was C57Bl/6 < CD1 = CF1 (P < 0.05, 1-way ANOVA). Across all three strains, exposure to > 320 pM peptide increased relative binding of sperm (P < 0.05; 2-way general linear model; GLM). Strains differed both in basal binding ability and in response to synthetic peptide. To determine if IVF rate increased after exposure of sperm to peptide, ova from B6C3 mice (four replicate pools) were collected after eCG and hCG stimulation. Cumulus-oocyte complexes (COC; 8-15 ova in each of 3-6 drops/treatment) were incubated with hyperactivated C57Bl/6 sperm at approximately 1500 sperm per ovum. Data for incubations were corrected for false-positive classification to yield a better estimate of true cleavage rate, and then related to results observed with a tenfold greater sperm concentration. Relative cleavage rates were 0 peptide (0.48); 420 pM (0.78, P < 0.05); and 840 pM (0.90, P < 0.01; GLM and Tukey tests). IVF rate was increased by exposure of mouse sperm to peptide at concentrations effective in the in vitro assay, and use of peptide allowed use of 1/10 as many sperm. PMID- 11066071 TI - Structure of the MLT gene and molecular characterization of the genomic breakpoint junctions in the t(11;18)(q21;q21) of marginal zone B-cell lymphomas of MALT type. AB - The t(11;18)(q21;q21) between the inhibitor of apoptosis API2 and the MLT gene is a distinct feature of marginal zone B-cell lymphomas of MALT-type. Hitherto the chimeric API2-MLT transcripts are all "in-frame" and predominantly fuse exon 7 of API2 to different MLT exons. Recurrent chromosomal translocations are common in lymphoid neoplasms and might represent by-products of the rearrangement processes generating antigen receptor diversity. The genomic structure of the MLT gene was determined to facilitate amplification of the genomic breakpoint junctions from 5 MALT-type lymphomas with t(11;18). Their sequence analysis showed scattering of the chromosome 11 breakpoints in intron 7 of API2 whereas rearrangements in MLT occurred in intron 2, 4, 7, or 8, respectively. Sequences around the junctions did not display recognition signal sequences mediating lymphocytic V(D)J recombination or other sequence motifs associated with recombination. The breakpoints occurred in a copy of an AluSx repeat in three cases, but interchromosomal Alu-mediated homologous recombination could be ruled out as the repeat resided only on one of the participating chromosomes. The t(11;18) was associated with a deletion in 4 out of 5 cases, ranging in size from 53 bp up to more than 200 kb. These deletions were observed on one or sometimes both derivative chromosomes that might indicate the susceptibility of these regions for breakage. Our data suggest that the API2-MLT fusion might result from a non homologous end joining event after multiple double-strand breaks. The clustering of breaks in intron 7 of API2 and the consistent "in frame" API2-MLT fusions could therefore reflect certain functional constraints crucial for clonal outgrowth. PMID- 11066072 TI - Low frequency of allelic imbalance at the prostate cancer susceptibility loci HPC1 and 1p36 in Swedish men with hereditary prostate cancer. AB - The aim of this study was to investigate allelic imbalance at the major human prostate cancer susceptibility locus HPC1 at 1q24-25 and the recently reported, putative, susceptibility locus at 1p36 in prostate tumors from Swedish families with hereditary prostate cancer. We analyzed 31 prostate tumors and two lymph node metastases from 33 Swedish men in 22 families with hereditary prostate cancer for the presence of allelic imbalance using microsatellite markers D1S158, D1S422, and D1S238 for the HPC1 locus and D1S1597, D1S407, and D1S489 for the 1p36 locus. Frequencies of allelic imbalance at the two investigated loci were quite low, 3 of 27 informative tumors at the 1p36 locus and 3 of 27 informative tumors at the HPC1 locus. Interestingly, two tumors showed allelic imbalance at both loci investigated, suggesting that they may have lost a great part of chromosome 1. Taking this possibility into consideration, the specific loss of the two investigated loci may be even lower (1 of 27 informative tumors for either locus). The very low level of allelic imbalance found at HPC1 and 1p36 makes it unlikely that these loci encode genes that are acting as classic tumor suppressor genes in the initiation or progression of hereditary prostate cancer. Of the eight tumors from HPC1-linked families, only two showed AI at the HPC1 locus, one of which had lost the wild-type allele. PMID- 11066073 TI - Topology of double minutes (dmins) and homogeneously staining regions (HSRs) in nuclei of human neuroblastoma cell lines. AB - Amplification of the MYCN gene is a characteristic feature of many neuroblastomas and is correlated with aggressive tumor growth. Amplicons containing this gene form either double minutes (dmins) or homogeneously staining regions (HSRs). To study the nuclear topology of these tumor-specific and transcriptionally active chromatin structures in comparison to chromosome territories, we performed fluorescence in situ hybridization with a MYCN probe and various chromosome paint probes, confocal laser scanning microscopy, and quantitative three-dimensional image analysis. The dmins formed dot-like structures in interphase nuclei and were typically located at the periphery of complexly folded chromosome territories; dmins noted in the chromosome territory interior were often detected within an invagination of the territory surface. Interphase HSRs typically formed extremely expanded structures, which we have never observed for chromosome territories of normal and tumor cell nuclei. Stretches of HSR-chromatin often extended throughout a large part of the cell nucleus, but appeared well separated from neighboring chromosome territories. We hypothesize that dmins are located within the interchromosomal domain (ICD) space and that stretches of HSR chromatin align along this space. Such a topology could facilitate access of amplified genes to transcription and splicing complexes that are assumed to localize in the ICD space. PMID- 11066074 TI - New comprehensive denaturing-gradient-gel- electrophoresis assay for KRAS mutation detection applied to paraffin-embedded tumours. AB - A comprehensive mutation detection assay is presented for the entire coding region and all splice site junctions of the KRAS oncogene. The assay is based on denaturing gradient gel electrophoresis and applicable to archival paraffin embedded tumour material. All KRAS amplicons are analysed within two lanes of a DGGE gel under a single set of experimental conditions. Six known codon 12 mutations in genomic DNA from different paraffin-embedded tumours could readily be detected. When testing 35 paraffin-embedded Dukes' C colorectal carcinomas for unknown mutations, 12 tumours were found with mutations in codons 12 or 13. None of the tumours appeared to have a codon 61 mutation. In nine tumours, however, 11 additional sequence variations were found outside the hot-spot codons. None of these occurred in germline DNA from 57 individuals. Of these variations, three are considered as significant mutations, as they result in a non-conservative substitution of amino acid residues essential for the functioning of the KRAS protein. Thus, in total, 15 of the 35 Dukes' C tumours (43%) had a KRAS mutation of functional significance. Moreover, a novel exon 4B polymorphism was found to occur in 10 of the 35 tumours. The results of this study suggest that in restricting analysis of KRAS to hot-spot mutation sites only, significant information may be missed. PMID- 11066075 TI - CD44 is a potential target of amplification within the 11p13 amplicon detected in gastric cancer cell lines. AB - Classical cytogenetic approaches have revealed many of the chromosomal aberrations that may occur in gastric cancers (GC), although few alterations of specific genes have been identified so far. Genes that affect progression of this disease need to be identified if clinicians are to achieve optimal management of patients with GC. As the first step toward the cloning of gene(s) that may be involved in gastric carcinogenesis, we examined 25 GC cell lines for aberrations in DNA copy number to detect chromosomal gains and losses, as well as gene amplifications, by comparative genomic hybridization (CGH). Our CGH study revealed high-level amplifications in chromosomal regions that had been well defined in GC but also in sites that had not, including 3p24, 5p15, 11p11.2-14, 13q34, 15q26, Xp24, and Xq26-28. The minimal common region at 11p13, within the 11p11.2-14 amplicon, harbors the CD44 gene. Northern and Western blot analyses showed that an alternatively spliced form of CD44, with variant exons 8-10 (CD44E), was overexpressed in all cell lines bearing the 11p13 amplicon. However, cell adhesion activity was no greater in these lines than in cell lines without amplifications at the CD44 locus, suggesting that the major property of upregulated CD44 in these cases might be to transduce signals critically associated with growth and proliferation of the tumor cells. PMID- 11066076 TI - t(7;12)(q36;p13), a new recurrent translocation involving ETV6 in infant leukemia. AB - The ETV6 gene is rearranged as a result of translocations involving a wide variety of chromosomal partners. To date, 12 partner genes for ETV6 have been cloned, and a further 23 chromosomal regions have been described. We previously identified a cryptic t(7;12) with ETV6 involvement in two cases of infant leukemia. The finding of a third case of t(7;12), also in an infant, prompted a more focussed search based on the common features found in these patients and those reported in the literature. The selection criteria were age at diagnosis < 20 months and the presence of +19 and/or +8 in the karyotype; cases with abnormalities of 7q and/or 12p were also considered. FISH studies using whole chromosome paints and probes for the ETV6 gene revealed a t(7;12) in 10 out of 23 cases studied. Seven of these had evidence of ETV6 rearrangement. Of those with ETV6 involvement, six had a 7q36 and one a 7q22 breakpoint. Importantly, in three cases the 7q36 breakpoint was within the same PAC, suggesting the existence of a new nonrandom translocation. However, in at least one patient the 7q36 breakpoint was different. The identification of the 7q partner genes will determine whether it is the disruption of ETV6 alone, or the formation of fusion genes, that is important for leukemogenesis in these patients. As both 7q36 and 7q22 are critical regions of gene loss in del(7q) leukemias, the identification of partner genes from these regions may also be important in understanding the pathogenesis of these diseases. PMID- 11066077 TI - Cytogenetic and molecular analysis of the acute monocytic leukemia cell line THP 1 with an MLL-AF9 translocation. AB - Cell lines derived from patients with leukemia are used in many molecular biology studies. Here we report the cytogenetic analysis of the THP-1 cell line using G banding, fluorescence in situ hybridization (FISH), and spectral karyotyping (SKY), and the molecular characterization of the MLL-AF9 rearrangement by RT-PCR. The THP-1 cell line was established from the peripheral blood of a 1-year-old boy with acute monocytic leukemia (AML-M5). THP-1 is near-diploid and consists of two related subclones with a number of aberrations, including the t(9;11), associated with AML M5. The use of FISH allowed us to identify and characterize otherwise hidden cytogenetic rearrangements, which include duplication of the 3' portion of MLL in the derivative 9 chromosome and a deletion of the 5' portion of the AF9 gene involved in the translocation. In addition to confirming the FISH results, SKY allowed for a more precise characterization of the karyotype of THP-1 and allowed us to identify other abnormalities in this cell line, including der(1)t(1;12), der(20)t(1;20), deletions 6p, 12p, and 17p, trisomy 8, and monosomy 10. Sequencing of the RT-PCR product showed a direct in-frame fusion product on the derivative chromosome 11 between exon 6 (exon 9) of MLL and exon 5 of AF9, which is most commonly involved in MLL-AF9 translocations. This study demonstrates that combining different techniques to achieve a more precise characterization of the THP-1 cell line provides important information that will be valuable for understanding the critical events required for leukemogenesis. PMID- 11066078 TI - Chromosome imbalances in familial gliomas detected by comparative genomic hybridization. AB - Familial occurrence of gliomas, in the absence of well-defined hereditary multisystem disorders, is reported occasionally. We describe 17 families that have been afflicted with two or more gliomas but do not raise suspicion of other inheritable syndromes. The families were identified among 369 consecutive glioma patients operated at the Tampere University Hospital during 1983-1994. We applied comparative genomic hybridization (CGH) analysis on 21 gliomas occurring in these 17 families. The most frequent genetic alterations, detected in over 20% of the tumors, were losses of 6q, 10, 4q, 9p and gains of 7, 19, 20q, 1p. We compared the chromosomal alterations detected in the familial gliomas to those reported previously on 209 sporadic gliomas in nine different CGH studies. In this comparison, the familial gliomas more often showed losses of chromosome arms 4q and 6q and gains of 1p and 22q. The most frequent losses (9/21 tumors) in the familial gliomas resided on chromosome arm 6q (P = 0.005, Fisher's exact test; with Bonferroni correction, P = 0.04). The loss of 6q was also the most common intrafamilial aberration, present in four separate gliomas belonging to two families. The minimal common area of loss on this chromosome resided at 6q14-16. In conclusion, we have found several characteristic aberrations by CGH in the familial gliomas and we present new chromosomal regions possibly involved in the familial predisposition to gliomas. PMID- 11066079 TI - Novel translocation of the BCL10 gene in a case of mucosa associated lymphoid tissue lymphoma. AB - Interest has focused on a recently identified gene, BCL10, thought to play an important role in the genesis of extranodal, marginal zone (MALT) lymphomas. This gene belongs to a family containing caspase recruitment domains (CARD), that are involved in the apoptotic pathway. Translocations of the BCL10 gene to the immunoglobulin heavy chain locus at 14q32 have been described. We report herein a case of MALT lymphoma showing t(1; 2)(p22; p12). The translocation was shown to involve the BCL10 gene and the immunoglobulin kappa light chain locus by fluorescence in situ hybridization. PMID- 11066080 TI - Deletion of 5q31 is observed in megakaryocytic cells in patients with myelodysplastic syndromes and a del(5q), including the 5q- syndrome. AB - One of the most common structural rearrangements in myelodysplastic syndrome (MDS) is a deletion of the long arm of chromosome 5, del(5q). The 5q- syndrome is a distinct entity, that presents with specific morphologic abnormalities of the megakaryocytic lineage. Thus, we evaluated the presence or absence of the del(5q) in these cells. We performed fluorescence in situ hybridization analysis using unique sequence probes (one for 5q31, the other for the 5p telomeric band), and tested bone marrow specimens from 10 patients with MDS (including 6 patients with the 5q- syndrome) and a del(5q). Megakaryocytes were identified by nuclear morphology, size, and ploidy index. Our results demonstrate the presence of the del(5q) in the megakaryocytic lineage and, thus, the involvement of these cells in the disease process. PMID- 11066081 TI - Inclusion of malignant fibrous histiocytoma in the tumour spectrum associated with hereditary non-polyposis colorectal cancer. AB - Sarcomas, including the malignant fibrous histiocytomas (MFHs), are not known to be part of the tumour spectrum of hereditary non-polyposis colorectal cancer (HNPCC) as epidemiologically established. Therefore, occurrence of MFH in an HNPCC family may very well be coincidental. HNPCC is associated with germline mutations in DNA mismatch repair genes, including the MSH2 gene. We analysed an MFH diagnosed in a 45-year-old male HNPCC patient carrying a germline MSH2 mutation for HNPCC-associated molecular characteristics, to investigate a possible relationship between the tumour and that mutation. DNA analysis revealed microsatellite instability and loss of one MSH2 copy, and immunohistochemistry showed absence of nuclear MSH2 protein staining. To investigate whether this is a common finding in MFH, microsatellite instability and nuclear MSH2 protein staining was tested for in 5 and 6 sporadic MFHs, respectively. None showed microsatellite instability and all stained positively for MSH2. Together, these findings show that in rare cases, MFH may be part of the HNPCC tumour spectrum. PMID- 11066082 TI - Expression and mutational analyses of the human MAD2L1 gene in breast cancer cells. AB - Breast cancer is a heterogeneous disorder in which most tumors display some degree of aneuploidy, especially those at later stages of the disease. Aneuploidy and associated chromosome instability may be important in the progression of mammary tumorigenesis. Aneuploidy is prevented during normal cell division in part through regulation of a mitotic spindle checkpoint where mitotic arrest prevents segregation of misaligned chromosomes into daughter cells at anaphase. Mitotic arrest genes, including the MAD family, which was originally characterized in yeast, help regulate normal function of the mitotic spindle checkpoint. Decreased expression of the human gene MAD2L1 was previously reported in a breast cancer cell line exhibiting chromosome instability and aneuploidy. To explore further the potential role of MAD2L1 in breast cancer, we analyzed MAD2L1 gene expression in 13 minimally to grossly aneuploid human breast cancer cell lines and found significant differences of expression in three lines. Sequence analysis of MAD2L1 cDNA in these as well as nine additional aneuploid breast cancer and five immortalized normal human mammary epithelial cell lines revealed one heterozygous frameshift (572 del A) mutation in a cancer cell line that demonstrated a high level of transcript expression. In addition, two 3'UTR sequence variants were noted in breast cancer cell lines. The 572 del A mutation creates a truncated MAD2 protein product. Further functional studies in primary breast tumors are therefore warranted to determine the potential role MAD2L1 may play in breast cancer. PMID- 11066083 TI - An identical HMGIC-LPP fusion transcript is consistently expressed in pulmonary chondroid hamartomas with t(3;12)(q27-28;q14-15). AB - The high frequency of the t(3;12)(q27-28;q14-15) in lipomas and pulmonary chondroid hamartomas (PCHs) makes the HMGIC-LPP fusion gene the most common fusion gene in a human tumor known so far. Nevertheless, there is no in-depth molecular analysis of the HMGIC-LPP fusion transcripts in PCHs. Certainly, a possible molecular variability of the HMGIC-LPP fusion may contribute to a better understanding of the histologic differences between lipomas and PCHs and the intratumoral histologic heterogeneity of PCHs. By RT-PCR and restriction analysis, we have investigated the HMGIC-LPP fusion transcripts in a series of 13 PCHs with t(3;12)(q27-28;q14-q15). HMGIC-LPP fusion transcripts of identical size were found in all PCHs tested. In all tumors investigated, the fusion transcripts had the same structure, i.e., exons 1 to 3 of HMGIC and exons 9 to 11 of LPP encoding a protein composed of three AT-hooks and two LIM-domains. Our results clearly show that neither the histologic differences between lipomas and PCHs nor the histologic heterogeneity of PCHs can be explained by a molecular diversity of the HMGIC-LPP fusion transcript. PMID- 11066084 TI - Pathological exon skipping in an HNPCC proband with MLH1 splice acceptor site mutation. AB - One of the most commonly mutated mismatch repair genes in human nonpolyposis colorectal cancer (HNPCC) is MLH1. We identified a splice site mutation in MLH1 in a colorectal cancer proband (T-to-A at position -11 of intron 1 splice acceptor) and investigated its functional consequences by RT-PCR, using lymphocyte mRNA from the proband, two noncarrying siblings, and one unrelated individual. Subcloning of PCR products followed by sequencing of individual clones revealed increased transcript heterogeneity in the mutation carrier, attributable to the presence of a variety of mRNA forms lacking exon 2, or combinations of exons 2, 4, 6, 9, and 10. The full-length transcript subcloned from the mutation carrier was detected with a much reduced frequency, suggesting that only the wild-type allele produced functional MLH1 mRNA. The three noncarriers expressed some previously described transcripts and several novel variants, but none that lacked exon 2. The results are consistent with the hypothesis that this splice site mutation causes skipping of MLH1 exon 2 in a large proportion of mRNA transcripts derived from the mutated allele. Such an observation strengthens the case for identifying the mutation as pathogenic in this HNPCC family, which is of interest given the rarity of exon skipping defects resulting from splice acceptor site mutations outside the invariant AG dinucleotide. PMID- 11066085 TI - Promoter demethylation accompanies reactivation of the HOX11 proto-oncogene in leukemia. AB - Despite considerable work on the epigenetic control of tumor suppressor genes, little is known about the potential role of promoter CpG demethylation in the activation of oncogenes in lymphoid tumors. The HOX11 proto-oncogene is frequently activated in T-cell acute lymphoblastic leukemia (T-ALL). HOX11 activation can occur in the absence of translocation of the gene to the T-cell receptor locus (Salvati et al., 1995), implying that activation mechanisms must be involved other than the juxtaposition of the gene to adjacent enhancing sequences. We tested whether the methylation status of the proximal promoter was correlated with expression status in T-ALL and found that, in all cases, expression of HOX11 in T-ALL was associated with extensive demethylation of the proximal HOX11 promoter, regardless of whether or not translocation was involved. In contrast, cells that did not express HOX11 showed a more methylated pattern of CpG residues in the proximal promoter. Methylation of this sequence in vitro was sufficient to silence the proximal promoter. We propose a model in which the selection of leukemia clones via a pathway involving HOX11 expression requires the demethylation of its promoter as a prerequisite for additional gene activation mechanisms. PMID- 11066086 TI - Redefining a critical region of LOH on 4p16.3 in bladder cancer. PMID- 11066087 TI - Somite development in zebrafish. AB - A full understanding of somite development requires knowledge of the molecular genetic pathways for cell determination as well as the cellular behaviors that underlie segmentation, somite epithelialization, and somite patterning. The zebrafish has long been recognized as an ideal organism for cellular and histological studies of somite patterning. In recent years, genetics has proven to be a very powerful complementary approach to these embryological studies, as genetic screens for zebrafish mutants defective in somitogenesis have identified over 50 genes that are necessary for normal somite development. Zebrafish is thus an ideal system in which to analyze the role of specific gene products in regulating the cell behaviors that underlie somite development. We review what is currently known about zebrafish somite development and compare it where appropriate to somite development in chick and mouse. We discuss the processes of segmentation and somite epithelialization, and then review the patterning of cell types within the somite. We show directly, for the first time, that muscle cell and sclerotome migrations occur at the same time. We end with a look at the many questions about somitogenesis that are still unanswered. PMID- 11066088 TI - Molecular and cellular biology of avian somite development. AB - Much of our understanding of early vertebrate embryogenesis derives from experimental work done with the chick embryo. Studies of the avian somite have played a key role in elucidating the developmental history of this important structure, the source of most muscle and bone in the organism. Here we review the development of the avian somite including morphological and molecular data on the origin of paraxial mesoderm, maturation of the segmental plate, specification and formation of somite compartments, and somite cell differentiation into cartilage and skeletal muscle. PMID- 11066089 TI - Associations of FGF-3 and FGF-10 with signaling networks regulating tooth morphogenesis. AB - The morphogenesis and cell differentiation in developing teeth is governed by interactions between the oral epithelium and neural crest-derived ectomesenchyme. The fibroblast growth factors FGF-4, -8, and -9 have been implicated as epithelial signals regulating mesenchymal gene expression and cell proliferation during tooth initiation and later during epithelial folding morphogenesis and the establishment of tooth shape. To further evaluate the roles of FGFs in tooth development, we analyzed the roles of FGF-3, FGF-7, and FGF-10 in developing mouse teeth. In situ hybridization analysis showed developmentally regulated expression during tooth formation for Fgf-3 and Fgf-10 that was mainly restricted to the dental papilla mesenchymal cells. Fgf-7 transcripts were restricted to the developing bone surrounding the developing tooth germ. Fgf-10 expression was observed in the presumptive dental epithelium and mesenchyme during tooth initiation, whereas Fgf-3 expression appeared in the dental mesenchyme at the late bud stage. During the cap and bell stage, both Fgf-3 and Fgf-10 were intensely expressed in the dental papilla mesenchymal cells both in incisors and molars. It is of interest that Fgf-3 expression was also observed in the primary enamel knot, a putative signaling center of the tooth, whereas no transcripts were seen in the secondary enamel knots that appear in the tips of future cusps of the bell stage tooth germs. Down-regulation of Fgf-3 and Fgf-10 expression in postmitotic odontoblasts correlated with the terminal differentiation of the odontoblasts and the neighboring ameloblasts. In the incisors, mesenchymal cells of the cervical loop area showed partially overlapping expression patterns for all studied Fgfs. In vitro analyses showed that expression of Fgf-3 and Fgf-10 in the dental mesenchyme was dependent on dental epithelium and that epithelially expressed FGFs, FGF-4 and -8 induced Fgf-3 but not Fgf-10 expression in the isolated dental mesenchyme. Beads soaked in Shh, BMP-2, and TGF-beta 1 protein did not induce either Fgf-3 or Fgf-10 expression. Cells expressing Wnt-6 did not induce Fgf-10 expression. Furthermore, FGF-10 protein stimulated cell proliferation in the dental epithelium but not in the mesenchyme. These results suggest that FGF-3 and FGF-10 have redundant functions as mesenchymal signals regulating epithelial morphogenesis of the tooth and that their expressions appear to be differentially regulated. In addition, FGF-3 may participate in signaling functions of the primary enamel knot. The dynamic expression patterns of different Fgfs in dental epithelium and mesenchyme and their interactions suggest existence of regulatory signaling cascades between epithelial and mesenchymal FGFs during tooth development. PMID- 11066090 TI - NF-kappa B is developmentally regulated during spermatogenesis in mice. AB - To analyze NF-kappa B activity in the testis, we used murine transgenic lines carrying a LacZ reporter gene under the control of a NF-kappa B-responsive promoter (Schmidt-Ullrich et al. [1996] Dev 122:2117-2128). We constructed three independent lines containing the promoter of the gene encoding p105, the precursor of the p50 subunit. This promoter contains three NF-kappa B-binding sites in its proximal part. Our results show that in adult mice, the beta galactosidase activity which reflects nuclear NF-kappa B activity, is first detected in spermatocytes at the pachytene stage and remains activated in the following steps of germ cell differentiation and maturation. Using transgenic mice carrying a p105nlslacZ construct with the 3 NF-kappa B sites mutated in the p105 promoter, we found a significant reduction in the transgene activity, confirming the important role of NF-kappa B in the activation of the transgene. To confirm the stage of induction during spermatogenesis, we analysed the beta galactosidase activity in the testes from prepuberal mice in which cells synchrouneously enter meiosis. We detected the transgene activity at 18 days after birth, corresponding to the pachytene stage in spermatocytes. In nuclear extracts prepared from prepuberal mice, we found a peak of NF-kappa B DNA-binding activity made of p50 and p65 subunits at day 18 after birth, which remains high in the later stages. Further analysis showed that I kappa B alpha and beta, but not epsilon are expressed in the testes. Altogether, these data suggest that NF kappa B factors are stage specifically controlled and may play a role during the development of sperm cells. PMID- 11066091 TI - VEGF is deposited in the subepithelial matrix at the leading edge of branching airways and stimulates neovascularization in the murine embryonic lung. AB - We used whole lung cultures as a model to study blood vessel formation in vitro and to examine the role that epithelial-mesenchymal interactions play during embryonic pulmonary vascular development. Mouse lungs were isolated at embryonic day 11.5 (E11.5) and cultured for up to 4 days prior to blood vessel analysis. Platelet endothelial cell adhesion molecule-1 (PECAM/CD31) and thrombomodulin (TM/CD141) immunolocalization demonstrate that vascular development occurs in lung cultures. The vascular structures identified in lung cultures first appear as a loosely associated plexus of capillary-like structures that with time surround the airways. To investigate the potential role of vascular endothelial cell growth factor (VEGF) during pulmonary neovascularization, we immunolocalized VEGF in embryonic lungs. Our data demonstrate that VEGF is uniformly present in the airway epithelium and the subepithelial matrix of E11.5 lungs. At later time points, E13.5 and E15.5, VEGF is no longer detected in the proximal airways, but is restricted to the branching tips of airways in the distal lung. RT-PCR analysis reveals that VEGF(164) is the predominant isoform expressed in lung cultures. Grafting heparin-bound VEGF(164) beads onto lung explants locally stimulates a marked neovascular response within 48 hr in culture. Semi quantitative RT-PCR reveals an 18% increase in PECAM mRNA in VEGF(164)-treated whole lung cultures as compared with untreated cultures. The restricted temporal and spatial expression of VEGF suggests that matrix-associated VEGF links airway branching with blood vessel formation by stimulating neovascularization at the leading edge of branching airways. PMID- 11066092 TI - Glypican-4 is an FGF2-binding heparan sulfate proteoglycan expressed in neural precursor cells. AB - FGF2 is a crucial mitogen for neural precursor cells in the developing cerebral cortex. Heparan sulfate proteoglycans (HSPGs) are thought to play a role in cortical neurogenesis by regulating the action of FGF2 on neural precursor cells. In this article, we present data indicating that glypican-4 (K-glypican), a GPI anchored cell surface HSPG, is involved in these processes. In the developing mouse brain, glypican-4 mRNA is expressed predominantly in the ventricular zone of the telencephalon. Neither the outer layers of the telencephalic wall nor the ventricular zone of other parts of the developing brain express significant levels of glypican-4, with the exception of the ventricular zone of the tectum. In cultures of E13 rat cortical precursor cells, glypican-4 is expressed in cells immunoreactive for nestin and the D1.1 antigen, markers of neural precursor cells. Glypican-4 expression was not detected in early postmitotic or fully differentiated neurons. Recombinant glypican-4 produced in immortalized neural precursor cells binds FGF2 through its heparan sulfate chains and suppressed the mitogenic effect of FGF2 on E13 cortical precursor cells. The spatiotemporal expression pattern of glypican-4 in the developing cerebral wall significantly overlaps with that of FGF2. These results suggest that glypican-4 plays a critical role in the regulation of FGF2 action during cortical neurogenesis. PMID- 11066093 TI - FGF5 stimulates expansion of connective tissue fibroblasts and inhibits skeletal muscle development in the limb. AB - FGF5 is expressed in the mesenchyme and skeletal muscle of developing and adult mouse limbs. However, the function of FGF5 during development of the limb and limb musculature is unknown. To elucidate the inherent participation of FGF5 during limb organogenesis, a retroviral delivery system (RCAS) was used to overexpress human FGF5 throughout developing hind limb of chicken embryos. Misexpression of the soluble growth factor severely inhibited the formation of mature myocytes. Limbs infected with RCAS-FGF5 contained smaller presumptive muscle masses as evidenced by a decrease in MyoD and myosin heavy chain expressing cells. In contrast, ectopic expression of FGF5 significantly stimulated proliferation and expansion of the tenascin-expressing, connective tissue fibroblast lineage throughout the developing limb. Histological analysis demonstrated that the increase in tenascin immunostaining surrounding the femur, ileum, and pubis in the FGF5 infected limbs corresponded to the fibroblasts forming the stacked-cell perichondrium. Furthermore, pulse labeling experiments with the thymidine analog, BrdU, revealed that the increased size of the perichondrium was attributable to enhanced cell proliferation. These results support a model whereby FGF5 acts as a mitogen to stimulate the proliferation of mesenchymal fibroblasts that contribute to the formation of connective tissues such as the perichondrium, and inhibits the development of differentiated skeletal muscle. These results also contend that FGF5 is a candidate mediator of the exclusive spatial patterning of the hind limb connective tissue and skeletal muscle. PMID- 11066094 TI - CD44 expression in the developing and growing rat intervertebral disc. AB - CD44 has been identified at the time of extracellular matrix formation and expansion in several sites of the developing embryo (Wheatley et al. [1993] Development 119:295-306). The nucleus pulposus, consisting of a hydrated extracellular matrix tissue at birth, not previously closely analyzed, was examined for expression of CD44 in the developing and aging rat intervertebral disc. CD44 was identified solely on notochordal cells from the first onset of intervertebral disc formation (day 15 embryo) through the loss of notochordal cells from the nucleus pulposus (12-24 months of age). No CD44 expression was found in the notochordal cells prior to disc formation or in any cells other than the notochordal cells in the annulus fibrosus or nucleus pulposus of the intervertebral disc. Using reverse transcriptase-polymerase chain reaction methodology, the single 365 amino acid CD44 standard, CD44s, open reading frame was amplified from notochordal cells isolated from the nucleus pulposus. Western blot analysis of a cultured nucleus pulposus notochordal cells total protein extract identified a single CD44 species devoid of chondroitin sulfate with a mass of approximately 85 kDa, characteristic of CD44s. Cell surface detection for CD44 was co-localized with hyaluronan and proteoglycans at first appearance of disc formation in the nucleus pulposus. PMID- 11066095 TI - Modulation of cell proliferation in the embryonic retina of zebrafish (Danio rerio). AB - We describe light-microscopically the development of the embryonic zebrafish eye with particular attention to cell number, cell proliferation, and cell death. The period from 16 to 36 hr post fertilization (hpf) comprises two phases; during the first (16-27 hpf) the optic vesicle becomes the eye cup, and during the second (27-36 hpf) the eye cup begins to differentiate into the neural retina and pigmented epithelium. All cells in the eye primordium are proliferative prior to 28 hpf, and the length of the cell cycle has been estimated to be 10 hr at 24-28 hpf (Nawrocki, 1985). Our cell counts are consistent with that estimate at that age, but not at earlier ages. A 10-hr cell cycle predicts that the cell number should increase by 7% per hr, but during 16-24 hpf the cell number increased by only 1.5% per hr. Despite the low rate of increase, all cells labeled with bromo deoxyuridine, so all were proliferative. We considered three possible explanations for the nearly-constant cell number in the first phase: proliferation balanced by cell emigration from the eye, proliferation balanced by cell death, and low proliferation caused by a transient prolongation of the cell cycle. We excluded the first two, and found direct support for the third. Previous examinations of the cell cycle length in vertebrate central nervous system have concluded that it increases monotonically, in contrast to the modulation that we have shown. Modulation of the cell cycle length is well-known in flies, but it is generally effected by a prolonged arrest at one phase, in contrast to the general deceleration that we have shown. PMID- 11066096 TI - Can gastric endoderm change the regionally specific inducing ability of presumptive small intestinal mesoderm? AB - This study was designed to establish the source of gut mesoderm's ability to induce regional pattern in the endoderm. The most obvious possibility is induction by the endoderm through epithelial-mesenchymal interaction. To test this experimentally, reciprocal quail/chick combinations were prepared of early proventricular endoderm (that is already known to be regionally determined) and presumptive small intestinal mesoderm. The combinations were cultured for 7 days to allow for 'programming' of the mesoderm by the endoderm. After removal of the proventricular endoderm the mesoderm was combined with young gizzard endoderm. It is known that gizzard endoderm can be provoked to develop in either a proventricular or a small intestinal direction by association with the appropriate mesoderm. Thus, by combining intestinal mesoderm 'programmed' by association with proventricular endoderm with gizzard endoderm, the subsequent differentiation of the gizzard endoderm would indicate whether or not the inducing ability of the intestinal mesenchyme had been altered. In addition to such experimental grafts, three types of control graft were prepared. The results of the experiment, based on the morphology of the grafts and the immunocytochemical analysis of selected endocrine cell types, showed that in the majority of cases the gizzard endoderm developed the features of small intestine, not those of proventriculus. This indicates that at the stages studied, endoderm does not act to program mesoderm with which it is associated. If this does occur, it must take place at an earlier stage, i.e., before the time of explantation of the presumptive small intestinal mesoderm (1.25 days of incubation). PMID- 11066097 TI - Mouse Nov gene is expressed in hypaxial musculature and cranial structures derived from neural crest cells and placodes. AB - NOV is a member of an emerging family of proteins, the CCN family, implicated in the control of cell growth and differentiation. During mouse development Nov is expressed predominantly in the skeletal and visceral muscles and in the nervous system. Transcripts are first detected in muscle precursor cells from 10.0 dpc and later in the hypaxial muscles of the trunk and shoulder/hip, as well as in the muscles of the head and in the smooth muscle of major vessels. In the nervous system, Nov is observed in the somatic motor neurons of the spinal cord from 12.5 dpc and in cranial structures derived either from neural crest cells or placodes, including V, VII, VIII, and IX ganglia and olfactory neuroepithelia. PMID- 11066098 TI - From Grenoble to Grenoble (1972-2000): limb development and regeneration revisited. PMID- 11066099 TI - Editorial PMID- 11066100 TI - The impact of the stent era on the management strategy for acute myocardial infarction: A population-based perspective. AB - We determined trends in the use of invasive diagnostic and revascularization strategies from a multihospital community-wide perspective for patients suffering acute myocardial infarction (AMI). Comparing 3,824 patients treated in the prestent era (1986-1993) to 1,915 patients hospitalized during the stent era (1995-1997), there was a significant increase in the use of invasive procedures and revascularization techniques across a broad spectrum of AMI patients during their index hospitalization. This resulted in a higher-risk profile of patients referred for invasive management of AMI in the stent era. Cathet. Cardiovasc. Intervent. 51:255-258, 2000. PMID- 11066101 TI - Radiation exposure of patients and coronary arteries in the stent era: A prospective study. AB - Previous studies have investigated the radiation dose to doctors and patients during coronary angiography and angioplasty, but most of them were retrospective, conducted in the prestent era, and results have not been consistent. Effective dose of 57 patients undergoing coronary angiography and/or angioplasty was assessed by using a dose-area product (DAP) to effective dose conversion factor. Radiation exposure risks to patients were then calculated for each procedure. Thermoluminescent dosimeters, mounted on a specially designed catheter that was advanced to the left or right sinus of Valsalva, were used to measure the dose received by the coronary arteries. Mean effective dose received by patients were 5.0 +/- 0.5 mSv for coronary angiography, 6.6 +/- 1.0 mSv for angioplasty, 10.2 +/- 1.5 mSv for angioplasty followed by stent implantation, 13.6 +/- 2.5 mSv for angiography followed by ad hoc angioplasty, and 16.7 +/- 2.8 mSv for angiography followed by ad hoc angioplasty and stent implantation. Patient risk of developing cancer after each procedure was 0.025%, 0.033%, 0.051%, 0.068%, and 0.084%, respectively. Corresponding mean coronary irradiation doses were 24 +/- 2.5, 31.0 +/- 3.6, 43.6 +/- 7.2, 55.0 +/- 7.5, and 64.7 +/- 5.6 mGy, respectively. A linear relationship of the DAP and the dose at the coronary arteries was found: DAP = 1,273 (cm(2)) x coronary dose (mGy). Radiation exposure to coronary arteries and associated risk to patients are relatively low, even following complicated, multivessel angioplasty with stent implantation. Our method can be used for calculation of radiation risk to patients and radiation dose to coronary arteries by using external dosimeters. Cathet. Cardiovasc. Intervent. 51:259-264, 2000. PMID- 11066102 TI - How much is too much? PMID- 11066103 TI - Lesion-to-lesion relationship of the restenosis process after placement of coronary stents. AB - The interrelation between multiple stented lesions within one patient in the restenosis process is only partially understood. From 492 patients with follow-up angiograms after coronary stent placement, 115 patients underwent multilesion procedures involving 233 lesions. In randomly chosen 39 patients with 79 lesions, additional intravascular ultrasound (IVUS) studies were performed to measure intimal hyperplasia cross-sectional area (CSA) and thickness. A general linear model with intraclass correlation was used to calculate the coefficient of correlation rho (rho = 1.0 indicates perfect correlation; rho = 0.0 indicates no correlation) of late loss, late loss index, intimal hyperplasia CSA, and intimal hyperplasia thickness. Multivariate analysis showed restenosis in the companion lesion (odds ratio 4.68, 95% confidence interval 1. 68-12.92, P = 0.003) and small minimal lumen diameter preintervention (odds ratio 0.28, 95% confidence interval 0.11-0.73, P = 0.009) to be predictors of restenosis. There was a weak correlation between multiple lesions within the same patient for late lumen loss rho = 0.28 (95% confidence interval 0.10-0.46, P < 0. 001) and late loss index. IVUS analysis demonstrated correlation of intimal hyperplasia CSA rho = 0.40 (95% confidence interval 0.06-0. 74, P = 0.009) and of intimal hyperplasia thickness. In conclusion, late loss and intimal hyperplasia demonstrate a correlation between multiple stented lesions within one patient. In addition to known lesion related factors, restenosis in a companion lesion is a predictor for restenosis. Cathet. Cardiovasc. Intervent. 51:266-272, 2000. PMID- 11066104 TI - Primary stenting in nonselected patients with acute myocardial infarction: the Multilink Duet in Acute Myocardial Infarction (MIAMI) trial. AB - Most randomized trials comparing primary stenting with primary coronary angioplasty (PTCA) excluded patients at high risk from enrollment, thus arising the important question about the generalizability of the randomized trial results to all patients with AMI. The aim of this study was to assess the feasibility and effectiveness of a primary infarct-related artery (IRA) stenting strategy using a second-generation tubular stent in nonselected patients with acute myocardial infarction (AMI). All patients with AMI were considered eligible for primary IRA stenting. No restriction was made based on age or clinical status on presentation, or coronary anatomy, except in cases of a reference IRA diameter < 2.5 mm. The primary endpoint of the study was clinical target vessel failure defined as death, reinfarction, or repeat TVR due to restenosis or reocclusion of the IRA. Between June 1998 and March 1999, 201 consecutive patients with AMI underwent mechanical recanalization of the IRA. The mean age was 64 +/- 12, and 16% of patients were aged 75 years or over. The incidence of shock was 9%. Primary IRA stenting was performed in 89% of the patients. Patients who underwent PTCA alone had a smaller IRA diameter as compared to patients with a stented IRA (2.48 +/- 0.46 mm vs. 3.15 +/- 0.37 mm; P < 0.001). There were no stent deployment failures. The 6-month primary endpoint rate was 15% (2 deaths, 27 repeat TVR, 0 reinfarctions), while the 6-month angiographic restenosis rate was 22%. Primary IRA stenting in nonselected patients with AMI is highly feasible and associated with favorable clinical and angiographic outcomes. Cathet. Cardiovasc. Intervent. 51:273-279, 2000. PMID- 11066105 TI - Stents for infarcts: the missing link? PMID- 11066106 TI - Coronary flow reserve is reflective of myocardial perfusion status in acute anterior myocardial infarction. AB - Our objective was to determine whether coronary vasodilatory reserve (CVR) correlates with the perfusion state of infarct zone in early recovery phase of acute anterior myocardial infarction (AMI). We studied 14 patients (11 males; mean age, 46 years) who had AMI and 6 control subjects who had chest pain but normal coronary angiograms. All patients underwent successful percutaneous revascularization of left anterior descending (LAD) coronary artery. Coronary flow velocity was measured using intracoronary (IC) Doppler at baseline and following IC injection of 18 microg of adenosine. Myocardial perfusion was evaluated by myocardial contrast echocardiography (MCE). CVR was higher in patients without a perfusion defect on MCE than in those with (2.48 +/- 0.21 vs. 1.66 +/- 0.13, P = 0.001). Subjects with a perfusion defect had a lower CVR than controls (1.66 +/- 0.13 vs.2.40 +/- 0.18, P < 0.05). CVR was > 2.0 in all subjects without a perfusion defect. There was a strong correlation between the magnitude of myocardial opacification in the LAD territory and CVR (r = 0.80, P < 0.01). Increase in peak diastolic flow velocity after adenosine infusion, but not systolic flow velocity, correlated with myocardial opacification index (r = 0.63, P = 0.016). CVR of infarct-related artery correlated closely with the perfusion status of the myocardium in infarct zone and those with a CVR > 2.0 had normal myocardial perfusion. These data suggest that CVR may be used to determine the perfusion state of the myocardium in the infarct zone, which is a known predictor of myocardial viability. Cathet. Cardiovasc. Intervent. 51:281-286, 2000. PMID- 11066108 TI - Radial is not radical, but is it practical? Confession of a nonuser. PMID- 11066107 TI - Transradial cardiac catheterization in elderly patients. AB - The safety and efficacy of transradial cardiac catheterization in elderly patients is unknown. This study examines procedure success rates for transradial catheterization in appropriately selected patients < 70 (n = 195) and >/= 70 (n = 83) years old. Elderly patients were less likely to be selected for the transradial approach (46% vs. 61%; P = 0.05). Although patients >/= 70 years old were more often female (39.7% vs. 24.1%; P = 0.008) and had a smaller body surface area (1.89 +/- 0.18 vs. 2.01 +/- 0.24 m2; P = 0. 001), procedure success rates did not differ (95.1% vs. 94.8%; P = NS). Procedure-related variables including procedure time (15.4 +/- 12.6 vs. 16.1 +/- 11.6 min; P = NS), amount of radiographic contrast (90.1 +/- 31.9 vs. 86.4 +/- 29.8 cc; P = NS), and number of catheters used (1.5 +/- 0.9 vs. 1.5 +/- 0.7; P = NS) were similar between groups. We conclude that transradial catheterization can be safely and effectively performed in selected elderly patients. Cathet. Cardiovasc. Intervent. 51:287 290, 2000. PMID- 11066109 TI - Proposed standards for clinical evaluation of patent ductus arteriosus occlusion devices. Multiorganization Advisory Panel to FDA for Pediatric Cardiovascular Devices. PMID- 11066110 TI - Transcatheter retrieval and repositioning of an Amplatzer device embolized into the left atrium. AB - A transcatheter technique is described for stabilization and retrieval of an embolized Amplatzer device, which was inadvertently deployed in the left atrium of a 2-year-old girl with hemodynamically significant atrial septal defect within the fossa ovalis. Since surgery was not available for immediate device retrieval, transcatheter means were chosen to prevent embolization of the device into the mitral valve. After stabilizing the device by creating a guidewire circuit through the wire mesh of the right atrial disk, the Amplatzer device was repositioned through the interatrial defect by snaring the microscrew. No residual shunting and perfect device position was recorded during Doppler echocardiography directly after the procedure as well as 1, 2, and 120 days after the intervention. Cathet. Cardiovasc. Intervent. 51:297-300, 2000. PMID- 11066111 TI - Transcatheter closure of fontan fenestrations using the Amplatzer septal occluder: initial experience and follow-up. AB - We have recently used the Amplatzer septal occluder to close Fontan fenestrations. Between June 1998 and December 1999, 13 patients underwent transcatheter occlusion of their Fontan fenestrations. Systemic blood flow decreased significantly without a concomitant decrease in pulmonary blood flow. All residual shunts detectable by oximetry were at sites separate from those into which occlusion devices were implanted. One patient developed severe tricuspid regurgitation following the procedure requiring surgical removal of the device. At the last follow-up, all patients were doing well clinically. There were no shunts detectable through or around the devices by echocardiography. Our experience indicates that the location of the fenestration within the Fontan baffle is critical to avoiding device interference with other intracardiac structures. The Amplatzer septal occluder offers an effective means of transcatheter closure of Fontan baffle fenestrations. Although more experience is needed, our current follow-up data suggest that long-term outcomes will be favorable. Cathet. Cardiovasc. Intervent. 51:301-304, 2000. PMID- 11066112 TI - Transcatheter occlusion of atrial baffle leak after mustard repair. AB - We present a case of transcatheter closure of an atrial baffle leak with significant systemic to pulmonary atrium shunt in a patient late after Mustard operation and pulmonary valvotomy for transposition of the great arteries. This procedure alleviated the need for reoperation in a high-risk symptomatic patient. Cathet. Cardiovasc. Intervent. 51:305-307, 2000. PMID- 11066113 TI - Coil occlusion of a femoral arteriovenous fistula. AB - A femoral arteriovenous fistula was discovered in a 17-mo-old child with congenital heart disease and prior femoral cardiac catheterization. The fistulous connection was clearly visible by angiography with vein compression, and the fistula was closed percutaneously using a Gianturco coil. Cathet. Cardiovasc. Intervent. 51:308-311, 2000. PMID- 11066114 TI - Balloon deflection technique: A method to facilitate entry of a balloon catheter into a deployed stent. AB - The entry of an angioplasty balloon into a coronary stent is occasionally difficult due to poorly expanded stent struts or calcified tissue blocking balloon passage. We describe a simple technique using a second guidewire and balloon to facilitate entry into the stent. Cathet. Cardiovasc. Intervent. 51:312 313, 2000. PMID- 11066115 TI - The tricks of re-crossing a deployed stent. PMID- 11066116 TI - Non-surgical extraction of right cardiac "thrombus in transit". AB - In two hemodynamically unstable patients, massive pulmonary embolism and free floating right cardiac thrombi were diagnosed. Thrombolytic therapy was contraindicated and surgical treatment was rejected. In these two cases, we describe a successful non-surgical, percutaneous extraction of mobile right cardiac thrombi. Cathet. Cardiovasc. Intervent. 51:316-319, 2000. PMID- 11066117 TI - Successful management of coronary artery perforation with helical platinum microcoil embolization. AB - A case of small coronary artery perforation during coronary intervention is presented. Continued leakage occurred despite prolonged intracoronary balloon inflation, in part probably related to the use of glycoprotein (GP) IIB/IIIA inhibitors. It was successfully managed by microcoil embolization without any sequel, helping avoid surgery in a high-risk patient. Cathet. Cardiovasc. Intervent. 51:320-322, 2000. PMID- 11066118 TI - Retinal cholesterol emboli during diagnostic cardiac catheterization. AB - Retinal embolism is a highly infrequent complication of cardiac catheterization of thrombotic, lipidic, and calcific etiology. We provide the first reported clinical case of retinal embolism caused by cholesterol crystal without systemic adverse effects as a severe complication of diagnostic cardiac catheterization. Cathet. Cardiovasc. Intervent. 51:323-325, 2000. PMID- 11066119 TI - Cholesterol embolization syndrome: unifying principles. PMID- 11066120 TI - Sinoatrial nodal artery aneurysm with right ventricular outflow tract compression: report of a case. AB - We described a 16-year-old boy with sinoatrial nodal (SAN) artery aneurysm that drained into right atrium and compressed right ventricular outflow tract. The patient was clinically asymptomatic. Hemodynamic study revealed a 15 mm Hg peak systolic pressure gradient at right ventricular outflow tract. The fistula was successfully excised without sequalae. Cathet. Cardiovasc. Intervent. 51:328-331, 2000. PMID- 11066121 TI - Ultrasound thrombolysis in stent thrombosis. AB - Recent refinement in stent implantation technique and peri-procedural pharmacological treatment has lowered the incidence of stent thrombosis significantly. Still, all stent thromboses are associated with major adverse events. In previous studies it has been suggested that intravascular ultrasound fibrinolysis is safe and effective. In this report, ultrasound successfully reperfused thrombotically occluded stents. These observations suggest that ultrasound may dissolve occlusive platelet-rich thrombus effectively and safely. Cathet. Cardiovasc. Intervent. 51:332-334, 2000. PMID- 11066122 TI - Intracoronary measurement of pulsus alternans. AB - Pulsus alternans is typically found in patients with left ventricular systolic dysfunction. We describe a woman with biventricular systolic dysfunction and pulsus alternans in the right ventricle, pulmonary artery, and aorta. Coronary angiography revealed an intermediate stenosis in the proximal left anterior descending (LAD) coronary artery. Pulsus alternans was demonstrated in the mid LAD using a 0.014" guidewire-mounted pressure sensor. An abnormal fractional flow reserve was measured in the LAD. Cathet. Cardiovasc. Intervent. 51:335-338, 2000. PMID- 11066123 TI - Carotid stent placement for extracranial carotid artery disease: current state of the art. AB - Percutaneous revascularization techniques have dramatically altered traditional approaches to the management of both coronary and peripheral vascular disease. Their major advantage is that they are less invasive than conventional surgical procedures, offering revascularization without the risk of general anesthesia and with lesser procedural morbidity and mortality, shorter hospital stay, and lower cost. In patients with comorbidities that increase their risk of surgical complications, percutaneous revascularization techniques are the procedures of choice. The Achilles heel of balloon angioplasty, the higher risk of lesion recurrence, restenosis, has been markedly reduced with the use of endovascular stents. Over the past 20 years, percutaneous angioplasty and stenting have become accepted alternatives to surgical revascularization of aortoiliac, renal, femoropopliteal, subclavian, brachiocephalic, and dialysis access lesions. The most recent application of percutaneous intervention has been to explore its clinical utility and safety for stroke prevention in stenotic extracranial carotid arteries. Cathet. Cardiovasc. Intervent. 51:339-346, 2000. PMID- 11066124 TI - The skirt technique: A stenting technique to treat a lesion immediately proximal to the bifurcation (pseudobifurcation). AB - We report a new stenting technique employed in 10 patients to treat lesions immediately proximal to a bifurcation (pseudobifurcation). A stent is mounted on two balloon catheters and advanced into the lesion immediately proximal to the bifurcation until the distal portion of the balloon catheters enters each branch at the bifurcation. This is followed by kissing balloon inflation for stent deployment. In all 10 cases, the final angiogram showed a good result at the lesion site and no ostial compromise of any of the branches. Cathet. Cardiovasc. Intervent. 51:347-351, 2000. PMID- 11066125 TI - Five French (5 Fr) guiding catheters for percutaneous coronary angioplasty and stent placement: An initial feasibility study. AB - Thirty patients were treated with percutaneous coronary intervention (PCI) using a 5 Fr guiding catheter. A recently developed, mechanically advantaged hand injector was used to deliver contrast and achieved excellent visualization through the 5 Fr system. Stent sizes ranged from 2.25 to 4.00 mm in diameter and from 8 to 24 mm in length. All primary lesions were successfully treated. The average contrast use was 70 cc per case. There were no major complications and only one minor femoral hematoma. In selected patients, a balloon angioplasty and stent placement can be performed safely and successfully with 5 Fr guiding catheters using currently available products. This technique creates a smaller arterial puncture site, which may obviate the need for a closure device and allow early and safe ambulation. With 5 Fr systems, it appears that contrast usage is reduced, thereby potentially decreasing cost and morbidity. Cathet. Cardiovasc. Intervent. 51:352-357, 2000. PMID- 11066126 TI - Selective regional myocardial infiltration by the percutaneous coronary venous route: A novel technique for local drug delivery. AB - Recent advances in the treatment of heart disease, in particular cardiovascular gene therapy and therapeutic angiogenesis, highlight the need for efficient and practical local delivery methods for the heart. We assessed the feasibility of percutaneous selective coronary venous cannulation and injection as a novel approach to local myocardial drug delivery. In anesthetized swine, the coronary sinus was cannulated percutaneously and a balloon-tipped catheter advanced to the anterior interventricular vein (AIV) or middle cardiac vein (MCV). During balloon occlusion, venous injection of radiographic contrast caused regional infiltration of targeted myocardial regions. Complete AIV occlusion had no impact on LAD flow parameters. Videodensitometric analysis following venous injection showed that radiographic contrast persisted for at least 30 min. Selective regional myocardial infiltration is feasible by this approach, targeting selected myocardial beds, including the apex, anterior wall, septum, and inferoposterior wall. This novel technique has potential application for local myocardial drug or growth factor delivery. Cathet. Cardiovasc. Intervent. 51:358-363, 2000. PMID- 11066127 TI - A novel platinum-iridium, potentially gamma radioactive stent: evaluation in a porcine model. AB - In-stent restenosis (ISR) is a major problem within stented arteries. Surface treatment of stents with platinum and gold were found to have the maximum charge with least neointima formation (NF). This study was designed to evaluate platinum (maximum electrical charge) as a material to make stents to reduce NF. Iridium was added to make an alloy suitable for stent manufacture, with the potential to make the stent radioactive. We implanted the novel platinum-iridium (PI) stent in 10 porcine coronaries and compared to the Palmaz-Schatz (PS) stent implanted in 8 coronary arteries. Six weeks after implantation, angiography of the stented vessel was performed before sacrifice. The coronaries were perfusion-fixed and stained, and vessel parameters were analyzed by computer-aided histomorphometry. The thrombus formation and the inflammatory response was less in the PI stent (0.04 +/- 0.1 vs. 0.24 +/- 0.2, P = 0.005; and 1.1 +/- 0.5 vs. 2.4 +/- 0.3, P < 0.001). The NF from PI-stented arteries was smaller in size than the PS controls (1.9 +/- 0.6 mm(2) vs. 2.4 +/- 0.4 mm(2), P = 0.06). However, PI stents presented with higher recoil than the PS stent (16% vs. 5%, P < 0.001). Platinum-iridium is a highly biocompatible material with high performance, low inflammatory response with small NF. This stent does not lead to thrombus formation and has the potential (due to the presence of iridium) to be irradiated to form a gamma radioactive stent. Cathet. Cardiovasc. Intervent. 51:364-368, 2000. PMID- 11066128 TI - Re: Morocutti et al.: Bail-out rotational atherectomy to ablate stent struts after treatment of a LAD bifurcation lesion with the Trousers technique. PMID- 11066129 TI - If you hear hoof beats in the backyard, look not for a zebra PMID- 11066130 TI - Liquid glue for vascular sealing: is Duett's composition safe and effective? PMID- 11066131 TI - Acute necrotizing pancreatitis: treatment strategy according to the status of infection. AB - OBJECTIVE: To determine benefits of conservative versus surgical treatment in patients with necrotizing pancreatitis. SUMMARY BACKGROUND DATA: Infection of pancreatic necrosis is the most important risk factor contributing to death in severe acute pancreatitis, and it is generally accepted that infected pancreatic necrosis should be managed surgically. In contrast, the management of sterile pancreatic necrosis accompanied by organ failure is controversial. Recent clinical experience has provided evidence that conservative management of sterile pancreatic necrosis including early antibiotic administration seems promising. METHODS: A prospective single-center trial evaluated the role of nonsurgical management including early antibiotic treatment in patients with necrotizing pancreatitis. Pancreatic infection, if confirmed by fine-needle aspiration, was considered an indication for surgery, whereas patients without signs of pancreatic infection were treated without surgery. RESULTS: Between January 1994 and June 1999, 204 consecutive patients with acute pancreatitis were recruited. Eighty-six (42%) had necrotizing disease, of whom 57 (66%) had sterile and 29 (34%) infected necrosis. Patients with infected necrosis had more organ failures and a greater extent of necrosis compared with those with sterile necrosis. When early antibiotic treatment was used in all patients with necrotizing pancreatitis (imipenem/cilastatin), the characteristics of pancreatic infection changed to predominantly gram-positive and fungal infections. Fine-needle aspiration showed a sensitivity of 96% for detecting pancreatic infection. The death rate was 1.8% (1/56) in patients with sterile necrosis managed without surgery versus 24% (7/29) in patients with infected necrosis (P <.01). Two patients whose infected necrosis could not be diagnosed in a timely fashion died while receiving nonsurgical treatment. Thus, an intent-to-treat analysis (nonsurgical vs. surgical treatment) revealed a death rate of 5% (3/58) with conservative management versus 21% (6/28) with surgery. CONCLUSIONS: These results support nonsurgical management, including early antibiotic treatment, in patients with sterile pancreatic necrosis. Patients with infected necrosis still represent a high-risk group in severe acute pancreatitis, and for them surgical treatment seems preferable. PMID- 11066132 TI - Pancreatic necrosis: to debride or not to debride-that is the question. PMID- 11066133 TI - The short esophagus: pathophysiology, incidence, presentation, and treatment in the era of laparoscopic antireflux surgery. AB - OBJECTIVE: To discuss the pathophysiology and incidence of the short esophagus, to review the history of treatment, and to describe diagnosis and possible treatments in the era of laparoscopic surgery. SUMMARY BACKGROUND DATA: The entity of the short esophagus in antireflux surgery is seldom discussed in the laparoscopic literature, despite its emphasis in the open literature for more than 40 years. This may imply that many laparoscopic patients with short esophagi are unrecognized and perhaps treated inappropriately. Intrinsic shortening of the esophagus most commonly occurs in patients with chronic gastroesophageal reflux disease that involves recurring cycles of inflammation and healing, with subsequent fibrosis. The actual incidence of the short esophagus is estimated to be approximately 10% of patients undergoing antireflux surgery. Of this group, 7% can be appropriately managed with extensive mediastinal mobilization of the esophagus to achieve the required esophageal length. The remaining 3% require an aggressive surgical approach, including the use of gastroplasty procedures, to create an adequate length of intraabdominal esophagus to perform a wrap. Several effective minimally invasive techniques have been developed to deal with the short esophagus. CONCLUSIONS: Because a short esophagus is uncommon, there is a natural concern that many surgeons will not perform enough antireflux procedures to become familiar with its diagnosis and management. A complete understanding of the short esophagus and methods for surgical correction are critical to avoid "slipped" wraps and mediastinal herniation and to achieve the best patient outcome. PMID- 11066134 TI - Early experience with laparoscopic approach for solid liver tumors: initial 16 cases. AB - OBJECTIVE: To evaluate the feasibility and outcome of laparoscopic hepatectomy in patients with solid liver tumors. SUMMARY BACKGROUND DATA: Although the laparoscopic approach has become popular in the surgical field, the value of laparoscopy in liver surgery is unknown. METHODS: Fifteen patients with solid liver tumors underwent 16 consecutive laparoscopic resections at the authors' institution between 1994 and 1999. Indications were symptomatic hemangioma, focal nodular hyperplasia, liver cell adenoma, isolated metastasis from a colon cancer, and hepatocellular carcinoma. The laparoscopic procedure was performed using four to seven ports (four 10-mm, two 5-mm, and one 12-mm). RESULTS: One patient underwent a major hepatic resection (right lobectomy); the others underwent minor hepatic resections (left lateral segmentectomies, IVb subsegmentectomies, segmentectomy, and nonanatomical excisions). The laparoscopic procedure was uneventful in 15 patients; one patient required conversion to open laparotomy because of inadequate free surgical margins. CONCLUSION: Laparoscopic surgery of the liver is feasible. The use of this new technical approach offers many advantages but requires extensive experience in hepatobiliary surgery and laparoscopic skills. The authors' experience suggests that laparoscopic procedures should be reserved for benign tumors in selected cases. Its application must be verified by further studies. PMID- 11066135 TI - Comparison of reg I and reg III levels during acute pancreatitis in the rat. AB - OBJECTIVE: To study alterations of serum levels of the pancreatic reg family of proteins in two models of acute pancreatitis. SUMMARY BACKGROUND DATA: The pancreatic reg family of proteins is expressed in the acinar pancreas. Reg I (pancreatic stone protein, PSP) and reg III (pancreatitis-associated protein, PAP) are induced after the onset of acute pancreatitis, and both have been proposed as potential markers of pancreatitis. METHODS: Pancreatitis was induced in rats by either retrograde infusion of sodium taurocholate or by direct trauma. Serum samples were obtained daily for 4 days after the procedure, and the animals were then killed. Twelve animals underwent sham procedure and six underwent daily analysis without surgery. Levels of reg I/PSP and reg III/PAP were estimated by enzyme-linked immunosorbent assay. RESULTS: Reg III/PAP levels increased significantly the first day after induction of both types of pancreatitis and rapidly returned to baseline in all survivors. Even animals who received retrograde infusion of saline showed a mild increase in reg III/PAP on the first day, whereas control animals that did not undergo surgery showed no variations. Reg I/PSP serum levels remained unchanged throughout all experimental periods. Postinjury reg III/PAP levels significantly correlated with severity of the pancreatic injury and animal survival; reg I/PSP levels did not. CONCLUSION: After induction of pancreatitis, serum levels of reg I and III protein differ significantly. Reg III/PAP levels are a sensitive marker of pancreatic injury and early in the disease may be a useful prognostic indicator for disease severity. PMID- 11066136 TI - Prognostic significance of entrapped liver cells in hepatic metastases from colorectal cancer. AB - OBJECTIVE: To correlate the microscopic finding of entrapped liver cells in hepatic metastases from colorectal cancer with outcome after hepatectomy. SUMMARY BACKGROUND DATA: Reliable histopathologic prognostic factors in resected liver metastases from colorectal cancer have not been identified. METHODS: Seventy-one patients undergoing radical hepatectomy for liver metastases were assigned to rare (n = 36) or frequent (n = 35) groups according to the microscopically observed frequency of hepatocyte entrapment in the tumor. RESULTS: Five-year survival rates after hepatectomy were 44. 4% for the rare group and 27.2% for the frequent group. Multivariate analysis using the Cox proportional hazards model by a stepwise method identified this morphologic variable as a significant independent prognostic factor. CONCLUSIONS: The finding of entrapped liver cells in metastases from colorectal cancer reflects the biologic activity of the tumor and may be a useful prognostic indicator. PMID- 11066137 TI - Surgical anatomy of the left lateral segment as applied to living-donor and split liver transplantation: a clinicopathologic study. AB - OBJECTIVE: To evaluate intrahepatic vascular and biliary anatomy of the left lateral segment (LLS) as applied to living-donor and split-liver transplantation. SUMMARY BACKGROUND DATA: Living-donor and split-liver transplantation are innovative surgical techniques that have expanded the donor pool. Fundamental to the application of these techniques is an understanding of intrahepatic vascular and biliary anatomy. METHODS: Pathologic data obtained from cadaveric liver corrosion casts and liver dissections were clinically correlated with the anatomical findings obtained during split-liver, living-donor, and reduced-liver transplants. RESULTS: The anatomical relation of the left bile duct system with respect to the left portal venous system was constant, with the left bile duct superior to the extrahepatic transverse portion of the left portal vein. Four specific patterns of left biliary anatomy and three patterns of left hepatic venous drainage were identified and described. CONCLUSIONS: Although highly variable, the biliary and hepatic venous anatomy of the LLS can be broadly categorized into distinct patterns. The identification of the LLS duct origin lateral to the umbilical fissure in segment 4 in 50% of cast specimens is significant in the performance of split-liver and living-donor transplantation, because dissection of the graft pedicle at the level of the round ligament will result in separate ducts from segments 2 and 3 in most patients, with the further possibility of an anterior segment 4 duct. A connective tissue bile duct plate, which can be clinically identified, is described to guide dissection of the segment 2 and 3 biliary radicles. PMID- 11066138 TI - Percutaneous portal vein embolization increases the feasibility and safety of major liver resection for hepatocellular carcinoma in injured liver. AB - OBJECTIVE: To assess the influence of preoperative portal vein embolization (PVE) on the long-term outcome of liver resection for hepatocellular carcinoma (HCC) in injured liver. SUMMARY BACKGROUND DATA: On an healthy liver, PVE of the liver to be resected induces hypertrophy of the remnant liver and increases the safety of hepatectomy. On injured liver, this effect is still debated. METHODS: During the study period, 10 patients underwent preoperative PVE and 19 patients did not before resection of three or more liver segments for HCC in injured liver (cirrhosis or fibrosis). PVE was performed when the estimated rate of remnant functional liver parenchyma (ERRFLP) assessed by computed tomographic scan volumetry was less than 40%. RESULTS: In all patients, PVE was feasible. There were no deaths or complications. The ERRFLP after PVE was significantly increased compared with the pre-PVE value. Liver resection was performed after PVE in 9 of 10 patients, with surgical death and complication rates of 0% and 45%, respectively. PVE increased the number of resections of three or more segments by 47% (9/19). Overall actuarial survival rates with or without previous PVE (89%, 67%, and 44% vs. 80%, 53%, and 53% at 1, 3 and 5 years, respectively) and disease free actuarial survival rates (86%, 64%, and 21% vs. 55%, 17%, and 17% at 1, 3, and 5 years respectively) after hepatectomy were comparable. CONCLUSION: With the use of PVE, more patients with previously unresectable HCC in injured liver can benefit from resection. Long-term survival rates are comparable to those after resection without PVE. PMID- 11066139 TI - Estradiol administration after trauma-hemorrhage improves cardiovascular and hepatocellular functions in male animals. AB - OBJECTIVE: To determine whether female sex steroids have any salutary effects on the depressed cardiovascular and hepatocellular functions following trauma and hemorrhage in male animals. SUMMARY BACKGROUND DATA: Studies indicate that gender difference exists in the immune and cardiovascular responses to trauma hemorrhage, and that male sex steroids appear to be responsible for producing immune and organ dysfunction, but it remains unknown if female sex steroids produce any salutary effects on the depressed cellular and organ functions in males following trauma and hemorrhage. METHOD: Adult male Sprague-Dawley rats underwent a midline laparotomy (i.e., trauma induction), and were bled to and maintained at a mean arterial pressure of 40 mmHg until 40% of the maximum bleed out volume was returned in the form of Ringer's lactate (RL). Animals were then resuscitated with RL at 4 times the shed blood over 60 minutes. 17beta-estradiol (50 microg/kg) or an equal volume of vehicle was injected subcutaneously 15 minutes before the end of resuscitation. The maximal rate of ventricular pressure increase or decrease (+/-dP/dtmax), cardiac output, and hepatocellular function (i.e., maximal velocity and overall efficiency of in vivo indocyanine green clearance) were assessed at 24 hours after hemorrhage and resuscitation. Plasma levels of interleukin (IL)-6 were also measured. RESULTS: Left ventricular performance, cardiac output, and hepatocellular function decreased significantly at 24 hours after trauma-hemorrhage and resuscitation. Plasma levels of IL-6 were elevated. Administration of 17beta-estradiol significantly improved cardiac performance, cardiac output, and hepatocellular function, and attenuated the increase in plasma IL-6 levels. CONCLUSION: Administration of estrogen appears to be a useful adjunct for restoring cardiovascular and hepatocellular functions after trauma-hemorrhage in male rats. PMID- 11066140 TI - Logistics and technique for procurement of intestinal, pancreatic, and hepatic grafts from the same donor. AB - OBJECTIVE: To assess a technique for simultaneous recovery of the intestine, pancreas, and liver from the same donor. SUMMARY BACKGROUND DATA: With the more frequent use of pancreatic and intestinal transplantation, a procurement procedure is needed that permits retrieval of both organs as well as the liver from the same cadaveric donor for transplantation to different recipients. It is believed by many procurement officers and surgeons, however, that this objective is not technically feasible. METHODS: A technique for simultaneous recovery of the intestine, pancreas, and liver was used in 13 multiorgan cadaver donors during a 26-month period, with transplantation of the organs to 33 recipients. The intestine was removed from 11 donors separately and in continuity with the pancreas in the other 2. Six additional pancreases were excised and transplanted separately. Thirteen livers were retrieved, one of which was discarded because of steatorrhea. Ten of the remaining 12 livers were transplanted intact; the other 2 were split in situ and used as reduced-size hepatic allografts in four recipients. RESULTS: None of the 11 intestinal, 6 pancreatic, 2 intestinal pancreatic, or 14 whole or partial liver allografts sustained serious ischemic injury or were lost as a result of technical complications. One liver recipient died 25 months after surgery of recurrent C virus hepatitis. The other 32 recipients had adequate allograft function with a mean follow-up of 8 months. CONCLUSION: It was possible using the described technique to retrieve intestine, pancreas, and liver allografts safely from the same donor and to transplant these organs to different recipients. PMID- 11066141 TI - Portal venous and enteric exocrine drainage versus systemic venous and bladder exocrine drainage of pancreas grafts: clinical outcome of 40 consecutive transplant recipients. AB - OBJECTIVE: To test the hypothesis that pancreas transplantation using the more physiologic method of portal venous-enteric (PE) drainage could be performed without compromising patient and graft outcome, compared with the standard method of systemic venous-bladder (SB) drainage. METHODS: Between November 1995 and November 1998, the authors prospectively followed up 20 consecutive patients with SB drainage followed by 20 consecutive patients with PE drainage. All patients underwent simultaneous pancreas-kidney transplantation, and all were immunosuppressed with antilymphocyte serum, cyclosporin, azathioprine, and steroids. RESULTS: The actuarial patient survival rate at 1 year was 95% in the SB group and 100% in the PE group. Death-censored kidney graft survival was 100% in both groups; pancreas graft survival was 95% in the SB group and 100% in the PE group. The mean initial hospital stay was 15 days for both groups. However, during the first 6 months after transplantation, the SB group required more medical day-unit visits, mostly for treatment of metabolic acidosis and dehydration. The incidence of urinary tract infections was similar in both groups. The incidence of cytomegalovirus infections was significantly less in the PE group. The incidence of acute rejection was 37% in the SB group and 15% in the PE group. Mean serum creatinine levels 6 months after transplantation were significantly lower in the PE group than in the SB group. Glycemic control was excellent in both groups, but fasting serum insulin levels were significantly lower in the PE group. CONCLUSIONS: The PE method of pancreas transplantation can be performed with excellent patient and graft outcomes. PMID- 11066142 TI - Simultaneous cadaver pancreas living-donor kidney transplantation: a new approach for the type 1 diabetic uremic patient. AB - OBJECTIVE: To review the authors' experience with a new approach for type I diabetic uremic patients: simultaneous cadaver-donor pancreas and living-donor kidney transplant (SPLK). SUMMARY BACKGROUND DATA: Simultaneous cadaver kidney and pancreas transplantation (SPK) and living-donor kidney transplantation alone followed by a solitary cadaver-donor pancreas transplant (PAK) have been the transplant options for type I diabetic uremic patients. SPK pancreas graft survival has historically exceeded that of solitary pancreas transplantation. Recent improvement in solitary pancreas transplant survival rates has narrowed the advantage seen with SPK. PAK, however, requires sequential transplant operations. In contrast to PAK and SPK, SPLK is a single operation that offers the potential benefits of living kidney donation: shorter waiting time, expansion of the organ donor pool, and improved short-term and long-term renal graft function. METHODS: Between May 1998 and September 1999, the authors performed 30 SPLK procedures, coordinating the cadaver pancreas transplant with simultaneous transplantation of a laparoscopically removed living-donor kidney. Of the 30 SPLKs, 28 (93%) were portally and enterically drained. During the same period, the authors also performed 19 primary SPK and 17 primary PAK transplants. RESULTS: One-year pancreas, kidney, and patient survival rates were 88%, 95%, and 95% for SPLK recipients. One-year pancreas graft survival rates in SPK and PAK recipients were 84% and 71%. Of 30 SPLK transplants, 29 (97%) had immediate renal graft function, whereas 79% of SPK kidneys had immediate function. Reoperative rates, early readmission to the hospital, and initial length of stay were similar between SPLK and SPK recipients. SPLK recipients had a shorter wait time for transplantation. CONCLUSIONS: Early pancreas, kidney, and patient survival rates after SPLK are similar to those for SPK. Waiting time was significantly shortened. SPLK recipients had lower rates of delayed renal graft function than SPK recipients. Combining cadaver pancreas transplantation with living-donor kidney transplantation does not harm renal graft outcome. Given the advantages of living-donor kidney transplant, SPLK should be considered for all uremic type I diabetic patients with living donors. PMID- 11066143 TI - Impact of race on the outcome of carotid endarterectomy: a population-based analysis of 9,842 recent elective procedures. AB - OBJECTIVE: To examine the influence of race and other potentially confounding variables on the outcome of carotid endarterectomy (CEA). SUMMARY: Previous studies have demonstrated that CEA is performed less frequently in black patients, although little attention has been focused on the influence of race on the outcome of surgery. METHODS: The Maryland Health Services Cost Review Commission database was reviewed to identify all elective CEA procedures performed in all nonfederal acute care hospitals in the state from 1990 through 1995 to examine the influence of race and other factors on the rates of in hospital complications, in-hospital stroke, length of stay, and total hospital charges. RESULTS: Carotid endarterectomy was performed in 9,219 (94%) white and 623 (6%) black patients during this period. The in-hospital stroke rate was 1.7% 3. 1% among black patients and 1.6% among white patients. Black patients had a longer length of stay and higher mean hospital charges than white patients. Multivariate logistic regression analysis identified black race as an independent risk factor for in-hospital stroke. Performance of CEA by a high-volume surgeon was protective for the combined occurrence of in-hospital stroke or death, and whites were more than twice as likely to undergo surgery performed by high-volume surgeons. Conversely, undergoing surgery in a low-volume hospital was associated with in-hospital stroke, and blacks were four times as likely to use low-volume hospitals. CONCLUSIONS: Black patients who underwent elective CEA in Maryland from 1990 to 1995 had an increased incidence of in-hospital stroke, a longer hospital stay, and higher hospital charges than whites. Black race was identified as an independent risk factor for in-hospital stroke, although the reasons for this influence of race on outcome are undefined. The authors' observations also suggest the possibility of limited access to optimal surgical care among blacks, and this issue warrants further study. PMID- 11066144 TI - Richter's hernia and Sir Frederick Treves: an original clinical experience, review, and historical overview. AB - OBJECTIVE: To describe the clinical recognition, pathology, and management of Richter's hernia and to review the relevant literature of the past 400 years. SUMMARY BACKGROUND DATA: The earliest known reported case of Richter's hernia occurred in 1598 and was described by Fabricius Hildanus. The first scientific description of this particular hernia was given by August Gottlob Richter in 1778, who presented it as "the small rupture." In 1887, Sir Frederick Treves gave an excellent overview on the topic and proposed the title "Richter's hernia." To his work-a cornerstone to modern understanding-hardly any new aspects can be added today. Since then, only occasional case reports or small series of retrospectively collected Richter's hernias have been published. METHODS: The authors draw on their experience with 18 prospectively collected cases treated in the ICRC Lopiding Hospital for War Surgery in northern Kenya between February and December 1998 and review the relevant literature of the past 400 years. RESULTS: The classic features of Richter's hernia were confirmed in all case studies of patients: only part of the circumference of the bowel is entrapped and strangulated in the hernial orifice. The involved segment may rapidly pass into gangrene, yet signs of intestinal obstruction are often absent. The death rate in the authors' collective was 17%. CONCLUSION: Richter's hernia is a deceptive entity whose high death rate can be reduced by accurate diagnosis and early surgery. Considering the increasing incidence at laparoscope insertion sites, awareness of this special type of hernia with its misleading clinical appearance is important and of general interest. PMID- 11066145 TI - Laparoscopic techniques preserve immune function during and after surgery. PMID- 11066146 TI - Relevant prognostic factors in gastric cancer. PMID- 11066147 TI - The introduction of appendiceal CT. PMID- 11066148 TI - The use of laparoscopic ultrasonography in the preoperative study of patients with colorectal liver metastases. PMID- 11066149 TI - Letter to the editor PMID- 11066150 TI - Using z scores as a measurement of institutional trauma care improvement. PMID- 11066151 TI - Letter to the editor PMID- 11066152 TI - Relation of surgical volume to outcome. PMID- 11066153 TI - Letter to the editor PMID- 11066154 TI - The costs of endovascular versus transabdominal abdominal aortic aneurysm (AAA) repair. PMID- 11066155 TI - Letter to the editor PMID- 11066156 TI - Adjuvant radiotherapy and 5-fluorouracil after curative resection of cancer of the pancreas and periampullary region. PMID- 11066157 TI - Adjuvant radiotherapy and 5-fluorouracil after curative resection of cancer of the pancreas and periampullary region. PMID- 11066158 TI - Letter to the editor PMID- 11066160 TI - JAMA 100 years ago: MEDICAL LIBRARIES IN SMALLER CITIES PMID- 11066159 TI - A piece of my mind: I have always been fascinated... PMID- 11066161 TI - JAMA 100 years ago: LAY USE OF THE CLINICAL THERMOMETER PMID- 11066162 TI - Patient safety: from research to practice. PMID- 11066163 TI - Tiny "surgeons" prove surprisingly effective. PMID- 11066164 TI - Vaccines pose no diabetes, bowel disease risk. PMID- 11066166 TI - The world in medicine: RVF strikes the middle east PMID- 11066165 TI - Genetic screening to offset adult disease. PMID- 11066167 TI - The world in medicine: breast cancer in europe PMID- 11066168 TI - The world in medicine: translating HIV care PMID- 11066169 TI - The world in medicine: osteoporosis goes unnoticed PMID- 11066170 TI - Assessment and management of pain. PMID- 11066172 TI - Race, parity, and gestational diabetes as risk factors for type 2 diabetes mellitus. PMID- 11066174 TI - Estimating the numbers of smoking-related deaths. PMID- 11066176 TI - Health risks of cigar smoking. PMID- 11066177 TI - Health risks of cigar smoking PMID- 11066178 TI - Long-term neuroendocrine effects of childhood maltreatment. PMID- 11066180 TI - Long-term neuroendocrine effects of childhood maltreatment PMID- 11066179 TI - Long-term neuroendocrine effects of childhood maltreatment. PMID- 11066181 TI - Creutzfeldt-Jakob disease in the United States: 1979-1998. PMID- 11066182 TI - Measuring underuse of necessary care among elderly Medicare beneficiaries using inpatient and outpatient claims. AB - CONTEXT: Continuing changes in the health care delivery system make it essential to monitor underuse of needed care, even for relatively well-insured populations. Traditional approaches to measuring underuse have relied on patient surveys and chart reviews, which are expensive, or simple single-condition claims-based indicators, which are not clinically convincing. OBJECTIVE: To develop a comprehensive, low-cost system for measuring underuse of necessary care among elderly patients using inpatient and outpatient Medicare claims. DESIGN: A 7 member, multispecialty expert physician panel was assembled and used a modified Delphi method to develop clinically detailed underuse indicators likely to be associated with avoidable poor outcomes for 15 common acute and chronic medical and surgical conditions. An automated system was developed to calculate the indicators using administrative data. SETTING AND SUBJECTS: A total of 345,253 randomly selected elderly US Medicare beneficiaries in 1994-1996. MAIN OUTCOME MEASURES: Proportion of beneficiaries receiving care, stratified by indicators of necessary care (n = 40, including 3 for preventive care), and avoidable outcomes (n = 6). RESULTS: For 16 of 40 necessary care indicators (including preventive care indicators), beneficiaries received the indicated care less than two thirds of the time. Of all indicators, African Americans scored significantly worse than whites on 16 and better on 2; residents of poverty areas scored significantly lower than nonresidents on 17 and higher on 1; residents of federally defined Health Professional Shortage Areas scored significantly lower than nonresidents on 16 and higher on none (P<.05 for all). CONCLUSIONS: This claims-based method detected substantial underuse problems likely to result in negative outcomes in elderly populations. Significantly more underuse problems were detected in populations known to receive less-than-average medical care. The method can serve as a reliable, valid tool for monitoring trends in underuse of needed care for older patients and for comparing care across health care plans and geographic areas based on claims data. JAMA. 2000;284:2325-2333. PMID- 11066183 TI - Antibody concentration and clinical protection after Hib conjugate vaccination in the United Kingdom. AB - CONTEXT: The schedule for Haemophilus influenzae type b (Hib) vaccination of infants in the United Kingdom consists of 3 doses given at 2, 3, and 4 months of age. Many countries include a fourth dose (booster) of Hib vaccine in the second year of life on the basis of declining Hib antibody concentrations after the primary series. Few data are available to show that this fourth dose is actually necessary. OBJECTIVE: To evaluate long-term clinical protection against Hib disease and Hib antibody concentrations following primary Hib vaccination without a booster dose. DESIGN, SETTING, AND SUBJECTS: Clinical protection study conducted between October 1992 and March 1999 in the United Kingdom, in which children developing invasive Hib disease despite vaccination in infancy with 3 doses of Hib conjugate vaccine were reported by pediatricians through an active, prospective, national survey. Separate antibody studies were conducted among 2 cohorts of children (n = 153 and n = 107) vaccinated at 2, 3, and 4 months of age with Hib conjugate vaccine and followed up to 43 and 72 months of age. MAIN OUTCOME MEASURES: Age-specific vaccine effectiveness, derived from the observed number of true vaccine failures after 3 Hib vaccine doses compared with the number of cases expected based on the age-specific rates of invasive Hib disease obtained prior to the introduction of Hib vaccines; and proportion of children in the 2 cohorts with Hib antibody concentrations of less than 0.15 and less than 1.0 microg/mL. RESULTS: Ninety-six true vaccine failures occurring after 3 vaccine doses were detected. During the study period, an estimated 4,368,200 infants in the United Kingdom received 3 doses of vaccine; therefore, the vaccine failure rate was 2.2 per 100,000 vaccinees (95% confidence interval, 1.8-2.7 per 100,000). Although vaccine effectiveness declined significantly after the first year of life (P<.001), it remained high until the sixth year of life (99.4% in children aged 5-11 months vs 97.3% in those aged 12-71 months). The proportion of cohorts 1 and 2 with anti-PRP antibody levels of less than 0.15 microg/mL increased between 12 and 72 months of age (6% at 12 months, 8% at 43 months, and 32% at 72 months; chi(2)(1) = 18.25; P<.001 for trend). CONCLUSIONS: Our results suggest that anti-PRP antibody levels and clinical protection against Hib disease wane over time after Hib vaccination at 2, 3, and 4 months of age without a booster dose at 2 years of age. The decline in clinical protection is minimal, however, suggesting that a booster dose of Hib vaccine following infant vaccination is not essential. JAMA. 2000;284:2334-2340. PMID- 11066184 TI - Effect of community-based interventions on high-risk drinking and alcohol-related injuries. AB - CONTEXT: High-risk alcohol consumption patterns, such as binge drinking and drinking before driving, and underage drinking may be linked to traffic crashes and violent assaults in community settings. OBJECTIVES: To determine the effect of community-based environmental interventions in reducing the rate of high-risk drinking and alcohol-related motor vehicle injuries and assaults. DESIGN AND SETTING: A longitudinal multiple time series of 3 matched intervention communities (northern California, southern California, and South Carolina) conducted from April 1992 to December 1996. Outcomes were assessed by 120 general population telephone surveys per month of randomly selected individuals in the intervention and comparison sites, traffic data on motor vehicle crashes, and emergency department surveys in 1 intervention-comparison pair and 1 additional intervention site. INTERVENTIONS: Mobilize the community; encourage responsible beverage service; reduce underage drinking by limiting access to alcohol; increase local enforcement of drinking and driving laws; and limit access to alcohol by using zoning. MAIN OUTCOME MEASURES: Self-reported alcohol consumption and driving after drinking; rates of alcohol-related crashes and assault injuries observed in emergency departments and admitted to hospitals. RESULTS: Population surveys revealed that the self-reported amount of alcohol consumed per drinking occasion declined 6% from 1.37 to 1. 29 drinks. Self-reported rate of "having had too much to drink" declined 49% from 0.43 to 0.22 times per 6-month period. Self reported driving when "over the legal limit" was 51% lower (0. 77 vs 0.38 times) per 6-month period in the intervention communities relative to the comparison communities. Traffic data revealed that, in the intervention vs comparison communities, nighttime injury crashes declined by 10% and crashes in which the driver had been drinking declined by 6%. Assault injuries observed in emergency departments declined by 43% in the intervention communities vs the comparison communities, and all hospitalized assault injuries declined by 2%. CONCLUSION: A coordinated, comprehensive, community-based intervention can reduce high-risk alcohol consumption and alcohol-related injuries resulting from motor vehicle crashes and assaults. JAMA. 2000;284:2341-2347. PMID- 11066185 TI - Association between cigarette smoking and anxiety disorders during adolescence and early adulthood. AB - CONTEXT: Cigarette smoking is associated with some anxiety disorders, but the direction of the association between smoking and specific anxiety disorders has not been determined. OBJECTIVE: To investigate the longitudinal association between cigarette smoking and anxiety disorders among adolescents and young adults. DESIGN: The Children in the Community Study, a prospective longitudinal investigation. SETTING AND PARTICIPANTS: Community-based sample of 688 youths (51% female) from upstate New York interviewed in the years 1985-1986, at a mean age of 16 years, and in the years 1991-1993, at a mean age of 22 years. MAIN OUTCOME MEASURE: Participant cigarette smoking and psychiatric disorders in adolescence and early adulthood, measured by age-appropriate versions of the Diagnostic Interview Schedule for Children. RESULTS: Heavy cigarette smoking (>/=20 cigarettes/d) during adolescence was associated with higher risk of agoraphobia (10.3% vs 1.8%; odds ratio [OR], 6.79; 95% confidence interval [CI], 1.53-30.17), generalized anxiety disorder (20.5% vs 3.71%; OR, 5.53; 95% CI, 1.84 16.66), and panic disorder (7.7% vs 0.6%; OR, 15.58; 95% CI, 2.31-105.14) during early adulthood after controlling for age, sex, difficult childhood temperament; alcohol and drug use, anxiety, and depressive disorders during adolescence; and parental smoking, educational level, and psychopathology. Anxiety disorders during adolescence were not significantly associated with chronic cigarette smoking during early adulthood. Fourteen percent and 15% of participants with and without anxiety during adolescence, respectively, smoked at least 20 cigarettes per day during early adulthood (OR, 0.88; 95% CI, 0.36-2.14). CONCLUSION: Our results suggest that cigarette smoking may increase risk of certain anxiety disorders during late adolescence and early adulthood. JAMA. 2000;284:2348-2351. PMID- 11066186 TI - Treatment of acute hypoxemic nonhypercapnic respiratory insufficiency with continuous positive airway pressure delivered by a face mask: A randomized controlled trial. AB - CONTEXT: Continuous positive airway pressure (CPAP) is widely used in the belief that it may reduce the need for intubation and mechanical ventilation in patients with acute hypoxemic respiratory insufficiency. OBJECTIVE: To compare the physiologic effects and the clinical efficacy of CPAP vs standard oxygen therapy in patients with acute hypoxemic, nonhypercapnic respiratory insufficiency. DESIGN, SETTING, AND PATIENTS: Randomized, concealed, and unblinded trial of 123 consecutive adult patients who were admitted to 6 intensive care units between September 1997 and January 1999 with a PaO(2)/FIO(2) ratio of 300 mm Hg or less due to bilateral pulmonary edema (n = 102 with acute lung injury and n = 21 with cardiac disease). INTERVENTIONS: Patients were randomly assigned to receive oxygen therapy alone (n = 61) or oxygen therapy plus CPAP (n = 62). MAIN OUTCOME MEASURES: Improvement in PaO(2)/FIO(2) ratio, rate of endotracheal intubation at any time during the study, adverse events, length of hospital stay, mortality, and duration of ventilatory assistance, compared between the CPAP and standard treatment groups. RESULTS: Among the CPAP vs standard therapy groups, respectively, causes of respiratory failure (pneumonia, 54% and 55%), presence of cardiac disease (33% and 35%), severity at admission, and hypoxemia (median [5th 95th percentile] PaO(2)/FIO(2) ratio, 140 [59-288] mm Hg vs 148 [62-283] mm Hg; P =.43) were similarly distributed. After 1 hour of treatment, subjective responses to treatment (P<.001) and median (5th-95th percentile) PaO(2)/FIO(2) ratios were greater with CPAP (203 [45-431] mm Hg vs 151 [73-482] mm Hg; P =.02). No further difference in respiratory indices was observed between the groups. Treatment with CPAP failed to reduce the endotracheal intubation rate (21 [34%] vs 24 [39%] in the standard therapy group; P =.53), hospital mortality (19 [31%] vs 18 [30%]; P =.89), or median (5th-95th percentile) intensive care unit length of stay (6.5 [1 57] days vs 6.0 [1-36] days; P =.43). A higher number of adverse events occurred with CPAP treatment (18 vs 6; P =.01). CONCLUSION: In this study, despite early physiologic improvement, CPAP neither reduced the need for intubation nor improved outcomes in patients with acute hypoxemic, nonhypercapnic respiratory insufficiency primarily due to acute lung injury. JAMA. 2000;284:2352-2360. PMID- 11066187 TI - Association of noninvasive ventilation with nosocomial infections and survival in critically ill patients. AB - CONTEXT: Invasive life-support techniques are a major risk factor for nosocomial infection. Noninvasive ventilation (NIV) can be used to avoid endotracheal intubation and may reduce morbidity among patients in intensive care units (ICUs). OBJECTIVE: To determine whether the use of NIV is associated with decreased risk of nosocomial infections and improved survival in everyday clinical practice among patients with acute exacerbation of chronic obstructive pulmonary disease (COPD) or hypercapnic cardiogenic pulmonary edema (CPE). DESIGN AND SETTING: Matched case-control study conducted in the medical ICU of a French university hospital from January 1996 through March 1998. PATIENTS: Fifty patients with acute exacerbation of COPD or severe CPE who were treated with NIV for at least 2 hours and 50 patients treated with mechanical ventilation between 1993 and 1998 (controls), matched on diagnosis, Simplified Acute Physiology Score II, Logistic Organ Dysfunction score, age, and no contraindication to NIV. MAIN OUTCOME MEASURES: Rates of nosocomial infections, antibiotic use, lengths of ventilatory support and of ICU stay, ICU mortality, compared between cases and controls. RESULTS: Rates of nosocomial infections and of nosocomial pneumonia were significantly lower in patients who received NIV than those treated with mechanical ventilation (18% vs 60% and 8% vs 22%; P<.001 and P =.04, respectively). Similarly, the daily risk of acquiring an infection (19 vs 39 episodes per 1000 patient-days; P =.05), proportion of patients receiving antibiotics for nosocomial infection (8% vs 26%; P =.01), mean (SD) duration of ventilation (6 [6] vs 10 [12] days; P =.01), mean (SD) length of ICU stay (9 [7] vs 15 [14] days; P =.02), and crude mortality (4% vs 26%; P =.002) were all lower among patients who received NIV than those treated with mechanical ventilation. CONCLUSIONS: Use of NIV instead of mechanical ventilation is associated with a lower risk of nosocomial infections, less antibiotic use, shorter length of ICU stay, and lower mortality. JAMA. 2000;284:2361-2367. PMID- 11066188 TI - Evaluating patients with arthritis of recent onset: studies in pathogenesis and prognosis. AB - Inflammatory synovitis of recent onset poses a diagnostic and prognostic challenge to primary care physicians and rheumatologists. A lack of understanding of the underlying etiologic and pathogenic processes limits the ability to distinguish forms of arthritis that follow a benign, self-limiting course from forms that proceed to an aggressive, erosive disease requiring intensive immunosuppressive therapy. It is estimated that between 30% and 40% of patients presenting with early synovitis have disease that remains unclassified. Using data from a cohort of patients with early synovitis and reviewing current literature, we discuss investigational approaches toward a new classification of patients with early synovitis. Although a lack of understanding of this heterogeneous clinical syndrome has led clinicians to take a largely empirical approach to treatment thus far, the evolving awareness of disease predisposition at a genetic level and the expanding ability to specifically manipulate biological pathways may ultimately change the approach to this clinical problem. JAMA. 2000;284:2368-2373. PMID- 11066189 TI - The continuing quest for measuring and improving access to necessary care. PMID- 11066190 TI - Noninvasive positive pressure ventilation in acute respiratory failure. PMID- 11066191 TI - The trouble with ethics: results of a national survey of healthcare executives. PMID- 11066192 TI - Values, ethics, and moral reasoning among healthcare professionals: a survey. PMID- 11066193 TI - Organizational ethics Canadian style. PMID- 11066194 TI - Narrative, ethics, and human experimentation in Richard Selzer's "Alexis St. Martin": the miraculous wound re-examined. PMID- 11066195 TI - Should it be mandated that an HEC review a physician's decision not to honor a patient's or surrogate's refusal of treatment? PMID- 11066196 TI - Contemplating suicide. PMID- 11066197 TI - Changing abortion policy in Turkey. PMID- 11066198 TI - Do profiles change the way doctors practice? PMID- 11066199 TI - Who pays when a health plan sinks? PMID- 11066200 TI - Disability management. It takes a pro to solve the puzzle. PMID- 11066201 TI - The 100 top hospitals. PMID- 11066202 TI - Vision coverage: not just a pair of specs. PMID- 11066203 TI - Healthy construction. PMID- 11066204 TI - DataWatch. Vote of confidence for job-based insurance. PMID- 11066205 TI - How to justify the investment. PMID- 11066206 TI - Checklist for a good contract for IT purchases. PMID- 11066207 TI - Market resource guide. PMID- 11066208 TI - Medical errors, D.C.'s new cause--will result be action, or just talk? PMID- 11066209 TI - State insurance commissioners weigh PBM regulations this month. PMID- 11066210 TI - Compensation monitor. Providers gain capitation ground. PMID- 11066211 TI - Obesity / an unchecked epidemic. The next big thing in prevention may be no farther than your waist. PMID- 11066212 TI - Disease management. Getting out from under drug companies' shadows. PMID- 11066213 TI - Tracking the tracker of health care's trends. Interview by Patrick Mullen. PMID- 11066214 TI - An evidence-based evaluation of percutaneous vertebroplasty. AB - BACKGROUND INFORMATION: Percutaneous vertebroplasty is a therapeutic, interventional radiologic procedure that involves injection of bone cement into a cervical, thoracic, or lumbar vertebral body lesion for the relief of pain and the strengthening of bone. This procedure only recently has been introduced, and is being used for patients with lytic lesions due to bone metastases, aggressive hemangiomas, or multiple myeloma, and for patients who have medically intractable debilitating pain resulting from osteoporotic vertebral collapse. FINDINGS: Results from two uncontrolled prospective studies and several case series reports, including one with 187 patients, indicate that percutaneous vertebroplasty can produce significant pain relief and increase mobility in 70 percent to 80 percent of patients with osteolytic lesions in the vertebrae from hemangiomas, metastases, or myeloma, or with osteoporotic compression fractures. In these reports, pain relief was apparent within one to two days after injection, and persisted for at least several months up to several years. While experimental studies and preliminary clinical results suggest that percutaneous vertebroplasty can also strengthen the vertebral bodies and increase mobility, it remains to be proven whether this procedure can prevent additional fractures in the injected vertebrae. In addition, the duration of effect is not known; there were no long-term follow-up data on most of these patients, and these data may be difficult to obtain and interpret in patients with an underlying malignant process, because disease progression may confound evaluation of the treatment effect. Complications were relatively rare, although some studies reported a high incidence of clinically insignificant leakage of bone cement into the paravertebral tissues. In a few cases, the leakage of polymer caused compression of spinal nerve roots or neuralgia. Several instances of pulmonary embolism were also reported. Although patient selection criteria have not been definitely established, percutaneous vertebroplasty is considered appropriate treatment for patients with vertebral lesions resulting from osteolytic metastasis and myeloma, hemangioma, and painful osteoporotic compression fractures if the following criteria have been met: o Severe debilitating pain or loss of mobility that cannot be relieved by correct medical therapy. o Other causes of pain, such as herniated intervertebral disk have been ruled out by computed tomography or magnetic resonance imaging. o The affected vertebra has not been extensively destroyed and is at least one third of its original height. o Radiation therapy or concurrent surgical interventions, such as laminectomy, may also be required in patients with compression of the spinal cord due to ingrowth of a tumor. CONCLUSIONS: Percutaneous vertebroplasty has only recently been introduced as a treatment for osteolytic lesions and osteoporotic compression fractures of the vertebrae, but early results are promising. Up to 80 percent of patients with pain unresponsive to correct medical treatment experience a significant degree of pain relief, and few serious complications have been reported. However, relatively few patients have undergone this procedure, and there are no data from controlled clinical trials or from studies with long-term follow-up. At the present time this procedure is still in the investigational stages, but may be appropriate for patients with no other reasonable options for medical treatment. PMID- 11066215 TI - Beating obesity begins by expanding tort reform. PMID- 11066216 TI - Managed care outlook. Employer-based coverage makes gains while gaining confidence of Americans. PMID- 11066217 TI - The top 10 ways the Internet is changing health care I.T. PMID- 11066218 TI - The Internet can make strategic I.T. planning precarious. PMID- 11066219 TI - Today's interactivity offers glimpse of tomorrow's Web sites. PMID- 11066220 TI - Will the common Web browser take over health care? PMID- 11066221 TI - Out with the old, in with the new. PMID- 11066222 TI - Virtual network: efficient and safe. PMID- 11066223 TI - The online ties that bind. PMID- 11066224 TI - Just the fax, ma'am. A Denver PPO automates treatment authorizations, speeding and standardizing the entire process. PMID- 11066225 TI - Providers show some backbone. PMID- 11066226 TI - Giving physicians a helping hand. PMID- 11066227 TI - CIOs learn new skills. PMID- 11066228 TI - Hard(ware) lessons. PMID- 11066229 TI - www.educatingpatients.com? PMID- 11066230 TI - California 2010: "top 10 trends" for millennium. PMID- 11066231 TI - California gunfight at the "Capitation Corral". PMID- 11066233 TI - How to contain costs when lengths of stay are low. PMID- 11066232 TI - California health care's "top 10" millennium megatrends. PMID- 11066234 TI - From worst to first: a physician success story. PMID- 11066235 TI - Indicator instrument panels: a key to survival? PMID- 11066236 TI - Center for Case Management CareMap Tool for deconditioned frail elders. PMID- 11066237 TI - Day rehab fills gap between inpatient, outpatient care. PMID- 11066238 TI - Here are steps for starting a day treatment center. PMID- 11066239 TI - What's a typical day in a day treatment center? PMID- 11066240 TI - Hospital-based units do well under revised PPS according to initial analysis using preliminary grouper. PMID- 11066241 TI - HCFA reports impact of hospital-post acute transfer policy, may consider expanding to all DRGs. PMID- 11066242 TI - OIG increases amount, scope of civil money penalties for fraud. PMID- 11066243 TI - Decline in Medicare SNF funding is almost double BBA intent, says study; industry calls on Congress to act. PMID- 11066244 TI - Analysis shows broad range of payment changes, data anomaly under proposed PPS refinements. PMID- 11066245 TI - What to look for during a compliance review: 10 common mistakes made by SNFs. PMID- 11066246 TI - Plan Websites reach out. PMID- 11066247 TI - Measuring progress in e-health. PMID- 11066248 TI - The Internet: new frontiers in service to the health care consumer. PMID- 11066249 TI - Toward comparable electronic patient medical record information. PMID- 11066250 TI - From health plan executive to Internet pioneer: an interview with Managed Care On Line's Clive Riddle. PMID- 11066251 TI - On your side: empowering people with HIV/AIDS. PMID- 11066252 TI - A time to sow: strengthening public programs. PMID- 11066253 TI - Provider profiling: doing it right. PMID- 11066254 TI - Risk management in a hostile environment. PMID- 11066255 TI - By the numbers. Financially vulnerable beneficiaries depend on Medicare HMOs. PMID- 11066256 TI - Where does health care reform go from here? An unchartered odyssey. PMID- 11066257 TI - Health care spending: can the United States control it? AB - Health care spending in the United States has continued to outpace the growth in national income and the growth in spending in other countries. And yet many Americans are without sufficient health care. Since the failure of national health care reform proposals put forward by the Clinton administration and others, the United States has had to look for other solutions to the problem of how to control spending in this sector. Can the new competitive approach of managed care succeed where other cost control measures of the past have failed? This chapter begins with an examination of the problems facing health care today, outlines recent trends in health care spending, and details reasons why spending is rising so rapidly at this time. The historical context of health care reform proposals and government attempts to control spending are described next and the reasons why some of these plans made no progress are explained. The health care payment systems of other industrialized nations that have seen some success in controlling costs are analyzed. Comparison of these systems with proposed plans for reforming the U.S. system provide insights and lessons for the United States. Finally, the chapter describes managed care and managed competition and makes the argument that managed care has the potential to respond to many of the health care spending problems facing the United States. However, more data on this subject are needed, and the authors call for a national monitoring entity to assess the progress of managed care in meeting the health care needs of the public. PMID- 11066258 TI - The new organization of the health care delivery system. AB - The U.S. health care system is restructuring at a dizzying pace. In many parts of the country, managed care has moved into third-generation models emphasizing capitated payment for enrolled lives and, in the process, turning most providers and institutions into cost centers to be managed rather than generators of revenue. While the full impact of the new managed care models remains to be seen, most evidence to date suggests that it tends to reduce inpatient use, may be associated with greater use of physician services and preventive care, and appears to result in no net differences either positive or negative with regard to quality or outcomes of care in comparison with fee-for-service plans. Some patients, however, tend to be somewhat less satisfied with scheduling of appointments and the amount of time spent with providers. There is no persuasive evidence that managed care lowers the rate of growth in overall health care costs within a given market. Further, managed care performance varies considerably across the country, and the factors influencing managed care performance are not well understood. Organized delivery systems are a somewhat more recent phenomenon representing various forms of ownership and strategic alliances among hospitals, physicians, and insurers designed to provide more cost-effective care to defined populations by achieving desired levels of functional, physician-system, and clinical integration. Early evidence suggests that organized delivery systems that are more integrated have the potential to provide more accessible coordinated care across the continuum, and appear to be associated with higher levels of inpatient productivity, greater total system revenue, greater total system cash flow, and greater total system operating margin than less integrated delivery forms. Some key success factors for developing organized delivery systems have been identified. Important roles are played by organizational culture, information systems, internal incentives, total quality management, physician leadership, and the growth of group practices. This chapter describes the growth and evolution of managed care and organized delivery systems, the research evidence regarding managed care and organized delivery systems, and the likely future organization of the health system in light of recent trends and evidence. It also highlights some of the more important public policy implications of the new health care infrastructure. PMID- 11066259 TI - Strategic issues for managing the future physician workforce. AB - Physician workforce issues were among the most hotly debated components of the recent national health care reform effort. What are the United States' goals for its physician workforce? Will market forces be adequate to achieve these goals, or will regulatory intervention be needed? This chapter provides public and private policymakers with a framework for arriving at reasonable conclusions about this important subcomponent of national health policy. Physician supply and requirements are discussed first. A picture of the current U.S. physician workforce is presented, together with details of its size and the physician-to population ratio. Future growth of the physician workforce is projected, and future requirements are discussed along with the potential for both surpluses and shortages in some areas. Graduate medical education, a crucial topic in this discussion, is covered. The issue of substitution of nonphysician providers for physicians is considered next, with special attention paid to the capabilities of nonphysician providers in performing certain tasks, as well as the productivity and cost-effectiveness questions involved. While the physician supply in the United States may be adequate overall, gaps in service and problems with access to services persist in many rural and inner-city areas. The geographic distribution of the physician workforce and the balance of subspecialists and generalists are addressed. Other topics of discussion include the need for greater minority representation in the physician workforce and the evolving role of the physician executive. Finally, this chapter ends with a wrap-up of policy considerations and themes central to the new delivery system of the twenty-first century. These themes include market forces versus regulation, cost containment and workforce cost-effectiveness, the global role of the United States, and nonfinancial barriers to access to care, as well as the impact of technology and the role of physician scientists. PMID- 11066260 TI - States and health care reform: the importance of program implementation. AB - The recent debate on national health care reform marked another case of policy being considered without reference to how--or even if--it could be implemented. The debate revolved around broad issues, such as universal versus partial coverage, mandatory versus voluntary alliances, and the respective roles of government and the market in health care. The ease or even the possibility of successful implementation was not an ingredient in evaluating proposals. The burden of making health care reform work falls to the states. Whether in response to national reform or in implementing their own programs, they must move from a general reform blueprint to an actual program that delivers services. The hands on role of the states in designing and operating programs makes their implementation duties both unavoidable and critical. This chapter explores implementation issues that should be considered an integral part of planning for health care reform, at both the federal and the state level. The chapter has two goals. First, it makes a case for altering the usual approach to designing reform and recommends paying attention to implementation early in the policy process, rather than treating it as an afterthough. Second, it is intended to help policymakers design implementable programs and anticipate pitfalls. To achieve these goals, it examines the state role in health care reform; state capacity to carry out this role; examples of state health care reform initiatives and lessons for implementation drawn from these efforts; and barriers to successful implementation. The chapter concludes with recommendations for policymakers. PMID- 11066261 TI - The role of quality measurement in a competitive marketplace. AB - Quality measurement is not a new idea. However, in recent years, several new trends have gained prominence: greater interest in publicly reported information on quality of care, access to care, and patient satisfaction; an increased focus on health plans and integrated systems of care rather than on institutional providers and practitioners as the unit of observation; wide adoption of the techniques of continuous quality improvement within the health care sector; increased use of clinical practice guidelines to improve care for a broad range of medical conditions; incorporation of computer technology into the clinical setting; and greater appreciation for health outcomes as a measure of quality of care. This chapter first reviews the changes in the medical landscape that have seeded these trends and the distinction between quality assurance and quality improvement. It then focuses on public policy concerns, in particular on the emergence of publicly disseminated information about quality of care, now often called "quality report cards." The major prototypes of these reports developed to date, the responses to quality reporting by different members of the delivery system, and the major criticisms of this approach are reviewed. The chapter concludes by predicting probable developments and the strategies most likely to move health care forward in a productive direction. PMID- 11066262 TI - The legal framework for effective competition. AB - Largely because of its indifference to spiraling costs, the professional domination model is being replaced by a market model based on competition among managed care plans and integrated delivery systems. In general, the more fully integrated previously competing providers become--for instance, by assuming financial risk together--the less legal risk is present, because of a decreased possibility of improper conspiratorial or collective behavior. Nevertheless, provider joint ventures and integrated delivery systems face a complex interaction of practical challenges and various legal and regulatory risks. This chapter explores ways in which laws involving fraud and abuse, self-referral, private inurement, corporate practice of medicine, Medicare reimbursement policy, and antitrust enforcement affect typical integrated delivery systems. From a legal standpoint, it might seem logical that the laws regulating health care providers would support and promote integration. A permissive legal environment to foster development of an integrated service network model assumes its development in a delivery system in which networks are at financial risk for the services provided. However, many of the laws and regulations governing integrated provider development were established at a time when joint ventures and other alliances were organizing in a predominantly fee-for-service environment and were generating significant increases in health care costs without producing demonstrable efficiencies or quality enhancements. The results is a fundamental inconsistency in government policy. The demand for collaboration by purchasers and legislatures does not necessarily cause the vast body of health care regulators to revise their concerns that many of the very collaborative activities being encouraged trigger potentially illegal acts and relationships. In a market model, the application of federal and state antitrust laws is especially important. In 1993 and 1994, the Department of Justice and the Federal Trade Commission jointly issued "Statements of Antitrust Enforcement Policy" in a number of areas of provider uncertainty. For integrated delivery systems, the primary focus of antitrust analysis is "market power." Systems without market power (i.e., the ability to force a purchaser to do something that the purchaser would not do in a competitive market) cannot harm consumers and should be free from serious antitrust risk. Where a network may have market power, its activities may be limited only if demonstrable anticompetitive effects outweigh the benefits of the efficiencies claimed by the new arrangement. The chapter concludes that vigorous antitrust enforcement may be required to promote market competition among integrated networks of providers and the managed care plans they serve. PMID- 11066263 TI - The changing environment for technological innovation in health care. AB - A distinguishing feature of American health care is its emphasis on advanced technology. Yet today's changing health care environment is overhauling the engine of technological innovation. The rate and direction of technological innovation are affected by a complex of supply- and demandside factors, including biomedical research, education, patent law, regulation, health care payment, tort law, and more. Some distinguishing features of technological innovation in health care are now at increased risk. Regulatory requirements and rising payment hurdles are especially challenging to small technology companies. Closer management of health care delivery and payment, particularly the standardization that may derive from practice guidelines and clamping down on payment for investigational technologies, curtails opportunities for innovation. Levels and distribution of biomedical research funding in government and industry are changing. Financial constraints are limiting the traditional roles of academic health centers in fostering innovation. Despite notable steps in recent years to lower regulatory barriers and speed approvals, especially for products for life threatening conditions, the Food and Drug Administration is under great pressure from Congress, industry, and patients to do more. Technology gatekeeping is shifting from hundreds of thousands of physicians acting on behalf of their patients to fewer, yet more powerful, managed care organizations and health care networks. Beyond its direct effects on adoption, payment, and use of technologies, the extraordinary buying leverage of these large providers is cutting technology profit margins and heightening competition among technology companies. It is contributing to unprecedented restructuring of the pharmaceutical and medical device industries, leading to unprecedented alliances with generic product companies, health care providers, utilization review companies, and other agents. These industry changes are already having considerable effects on investment patterns and the development, adoption, and use of new technologies. Until recently, new technologies that offered the prospect for health benefit, however, marginal or unproven, were paid for with little or no regard to cost. Technical wizardry alone no longer carries the day in health care. Today's health care market increasingly demands what other markets do--measurable improvements in benefits at acceptable costs--and innovators have begun to respond accordingly. Even so, certain key venues for health care innovation are at risk. PMID- 11066264 TI - The special health care needs of the elderly. AB - Interest in Medicare, the government's second largest social program after Social Security, reached a new high in 1995, not as part of health care reform, but as a vehicle for deficit reduction and because of a desire by Congress to restructure the program to encourage enhanced choice for beneficiaries and greater use of managed care. Medicaid, a major payer of long-term care and financer of coverage for low-income elderly, also is slated to undergo major restructuring in the next few years. As Congress and the nation debate the future of these key programs for older Americans, a number of critical issues deserve attention. Medicare's costs are very high--but not necessarily unreasonable in the face of the demands on health care services for this part of the population. And even with these high costs, a number of important gaps in coverage remain a problem for seniors. Deductibles and copayments are also high--especially for hospital and skilled nursing services. But pressure for change may well lead to higher, not lower, cost-sharing requirements. Medicare remains a largely fee-for-service program at a time when the national health care system is shifting increasingly to a managed care environment. Moving Medicare in that direction is one likely option for change. While it is desirable to have Medicare move in concert with the rest of the system, a number of issues stand in the way of an effortless move to managed care for the elderly. Moreover, coordination of long-term and acute care services may be even more challenging in such an environment. Medicaid covers long-term care services for older Americans, but only for those who have depleted most of their assets and income. Even when people do become eligible, Medicaid covers primarily institutional care. But little is likely to change this picture in the next few years, and private efforts through expansion of long-term care insurance will likely provide only a partial solution. PMID- 11066265 TI - Redefining private insurance in a changing market structure. AB - This discussion on likely changes and challenges for the health insurance industry over the coming decade assumes that significant national reform of health care financing for the privately insured population will not occur--or, if it does, that it will mirror the insurance market reforms that many states already have undertaken. First, the changes in private insurance coverage during the past several years are considered, with particular attention to the erosion of employer-based coverage and to the rising influence of public insurance programs--especially Medicaid--on the private insurance market. Next is a description of the changing web of state laws and regulations governing private health insurance. At this writing, virtually every state has enacted or is considering reforms of the small group market to limit what many perceive as unfair or destructive insurer practices and to set new ground rules for competition among insurance arrangements. The changing nature of private insurance contracts in the United States is considered next. Evolving from conventional fee-for-service contracts, private insurance is increasingly a complex mixture of capitation, partial capitation, and reinsurance of capitated arrangements. Finally, this chapter discusses three issues of increasing importance in shaping the marketplace for private insurers: (1) the federal preemption of states' regulatory authority over self-insured employer plans; (2) emerging state regulation to restructure competition in the health insurance and health care markets; and (3) the growing interest of both federal and state governments in medical savings accounts to finance health insurance and health care spending. PMID- 11066266 TI - Managed care for people with disabilities: caring for those with the greatest need. AB - Disability is discussed in terms of three categories: conditions that result from biomedical conditions and chronic, lifelong illnesses; role or social functioning difficulties that result from behavioral, developmental, or brain disorders; and conditions that limit physical functioning. The range and depth of services needed by the disabled result in higher costs of health care for this population. Because their service needs vary so widely, no single program can address all of the needs equally. Currently, no integrated public policy or program is specifically designed to serve people with disabilities. Rather, they are served by a range of programs that provide specific benefits (e.g., health, social services, and income). Section 1 of this chapter provides an overview on extending the concept of managed care to disabled populations. Special attention is paid to the financing of health care, the delivery of care, reforming the health care system, the cost-containment potential of managed care, and the need to align care with the nature of the individual disability. In sections 2 and 3, the current status of managed care for two special populations--children and the mentally ill--is discussed in greater detail. Section 2 addresses the characteristics of chronically ill and disabled children, public and private health insurance coverage of children with disabilities, other public programs for chronically ill children, and current directions and strategic choices for managed pediatric care. Section 3 describes the mentally ill and the system of providers that currently supplies care to them, offers some conclusions regarding how managed care is changing the policy debate in mental health care, assesses the key factors affecting policy choices in managed care, and considers prospects for the future shape of managed behavioral health care. PMID- 11066267 TI - Rationing health care: what it is, what it is not, and why we cannot avoid it. AB - The word "rationing" has come to play a central role in the national health policy debate. Alas, it is also one of the most misunderstood of words. Its injection into the debate has generated far more heat than light. This chapter reviews the definition of "rationing" preferred by the profession that takes as its task the study of how individuals and society respond to and deal with scarcity, namely, the economics profession. It will be shown that economists usually consider all limits on the distribution of a scarce good or services to be "rationing," whether that limit takes the form of a price barrier or some method of non-price allocation--for example, queues or allocation by lottery. To make a distinction between allocation through freely competitive markets and other forms of resource allocation, economists distinguish between "price rationing" and "non-price rationing." This is a meaningful distinction. Adoption of the economist's definition of "rationing" would greatly clarify the national health policy debate. Next, the discussion turns to the controversial proposition, commonly made by most economists and a handful of their allies in the medical profession, that an economically efficient health care system will inevitably engage in the pervasive withholding of services that may be sought by patients and their physicians, and that it will do so to enhance the quality and efficiency of the health care system overall. If managed competition lives up to its current billing, it will entail rationing of precisely that sort. Unfortunately, the individualist tradition of the United States, as it expresses itself in the tort system, may seriously hinder managed competition from achieving its stated goal. Finally, this chapter offers some conjectures on the approach to rationing likely to be taken by the United States health care system in the twenty-first century. It is argued that, far from having been inconclusive, the most recent congressional debate on health care reform actually gave official sanction to a three-tiered health system, with fairly severe rationing in the bottom tier and virtually none in the top tier. While such tiering has always been present in the U.S. health care system, the phenomenon has hitherto been treated as a blemish to be removed by government--now it will probably remain a permanent fixture. PMID- 11066268 TI - Targeting the patient population. PMID- 11066269 TI - The impetus behind diabetes disease management programs. Accreditation/quality versus the bottom line. PMID- 11066270 TI - Patient self-education and patient-centric care. PMID- 11066271 TI - Whose clinical guidelines? PMID- 11066272 TI - Two looming issues and their implications for diabetes management. PMID- 11066273 TI - Can we measure the financial effect of diabetes management on health plan costs? PMID- 11066274 TI - Diabetes management: what's the quality diagnosis? PMID- 11066275 TI - Applying the ABCs of E-care. PMID- 11066276 TI - Long-term care based on the On Lok model. PMID- 11066277 TI - Predicting risk reduction of coronary disease in patients who are glucose intolerant: a comparison of treatment with fenofibrate and other lipid-modifying agents. AB - A clinical outcomes model was developed to estimate the ability of fibrates and statins to reduce the relative and absolute risk of developing coronary heart disease (CHD) in patients with diabetes. A database was constructed, drawing on results from 83 applicable published studies. Risk factor outcomes reported by the model were chosen according to those found to be statistically significant in the data set parameters from the Framingham Heart Study. In the analysis, fenofibrate was compared with gemfibrozil (an earlier fibrate) and six 3-hydroxy 3-methylglutaryl coenzyme A (HMG-CoA) reductase-inhibitor or "statin" agents. Among the agents studied, only fenofibrate was found to significantly affect all six cardiovascular disease risk factors used in this analysis. For men with diabetes, the estimated absolute risk of developing CHD after five years of treatment was lowered equally and most effectively by fenofibrate, simvastatin, and atorvastatin, from 10% to 6%. For women with diabetes, fenofibrate and simvastatin reduced absolute risk from 9% to 6% over five years of treatment, while atorvastatin reduced risk from 9% to 6%. An accompanying cost comparison analysis found that reducing absolute risk of CHD in these patients would result in significant reductions in the medical expense of treating cardiac events, and that significant differences in the amount saved were driven by treatment choice. Fenofibrate, simvastatin, and atorvastatin produced the greatest long-term savings. PMID- 11066278 TI - The role of innovation in restructuring health care delivery. PMID- 11066279 TI - Primary care and the de facto mental health care system: improving care where it counts. AB - Only 28% of individuals suffering from psychiatric disorders seek care from mental health specialists. In this paper, the authors describe how the de facto PCP mental health care system gained ground when it seemed the financing and organizational structure of managed care would have predicted the opposite result. They argue that the new realities of mental health practice require new approaches to improving behavioral health treatment. These approaches, they believe, will maximize the benefit of care delivered in and accessed through the primary care office. PMID- 11066280 TI - How XML will extend the world of managed care: Part I--Standardizing data communications. PMID- 11066281 TI - An integrated data warehouse system: development, implementation, and early outcomes. AB - This paper describes a generic vision of global information flow and the development of an integrated data warehouse system, using clinical data on all patient encounters and administrative data on all operating transactions as part of an integrated health care system. This new integrated data warehouse system has been successfully used for multiple purposes, including patient care, health services research, resource utilization and feasibility studies. During 1999, core analyses included the electronic abstraction, aggregation, and analysis of data on over 400,000 patients. This approach to building a centralized data system comprised of multiple repositories efficiently meets a variety of individual and aggregate information needs, while reducing the need to create duplicate databases. PMID- 11066282 TI - Stats & facts. Workers' compensation issues and implications. PMID- 11066283 TI - Health care e-predictions and e-trends for the next ten years. PMID- 11066284 TI - Is vertical integration adding value to health systems? AB - Vertical integration is a concept used by health systems when attempting to achieve economies of scale, greater coordination of services, and improved market penetration. This article focuses on the actual outcomes of utilizing vertical integration in the health field and then compares these findings with those reported in other industries. This analysis concludes that this organizational model does not work particularly well in the health industry, as illustrated by health alliances' poor fiscal performance when they acquire physician practices or when they start their own HMO plans. PMID- 11066285 TI - Cost, efficacy, effectiveness, and value: a medical hierarchy of need. PMID- 11066286 TI - Adverse drug reactions resulting in ambulatory service utilization. PMID- 11066287 TI - Utilization of ambulatory care services caused by adverse effects of medications in the United States. AB - Using the 1996 national ambulatory care surveys for the United States, this study examined the utilization of ambulatory care services because of the adverse effects of medications. Of the visits to ambulatory settings, 0.31% were because of adverse effects of medications, representing an estimated 2.73 million visits annually or 1.03 visits per 100 persons. Most (88%) of the visits were to office based physicians, and the remainder were to hospitals. From both cost and quality perspectives, implementing strategies to prevent and/or effectively manage the adverse effects of medications could prove beneficial for payers and patients. PMID- 11066288 TI - How XML will extend the world of managed care: Part II. PMID- 11066289 TI - The management of bacterial infections of the upper respiratory tract in children: pharyngitis and otitis media. AB - Despite changes in the activities of infectious microorganisms, particularly variations in their prevalences in different parts of the body, in clinically defined diseases, and in their susceptibilities to antibiotics, the established first-line agents for the empiric treatment of otitis media and pharyngitis are still valid. Nevertheless, a number of the newer antimicrobials have become valuable as second- and third-line medications when the first-choice drugs fail or are contraindicated. In terms of fundamental principles, the question of whether pharyngitis in children warrants routine antibiotic therapy remains unresolved; in practical terms, antibiotic treatment should be considered routinely. PMID- 11066290 TI - Qui tam litigation: "whistleblower" lawsuits. AB - Qui tam lawsuits are increasingly being used in the fight against fraud in the managed care environment. Brought by private individuals, called relators, on behalf of the U.S. government, the lawsuits aim to recover funds obtained through the presentment of false claims. The relator may eventually collect between 15% and 30% of the funds recovered. This article provides an overview of qui tam litigation and its role in the government's war against fraud. PMID- 11066291 TI - Effect of postmenopausal hormone replacement therapy on dental outcomes: systematic review of the literature and pharmacoeconomic analysis. AB - A systematic review and economic analysis of clinical trials evaluating the effect of hormone replacement therapy (HRT) on dental outcomes in postmenopausal women were conducted. Twenty published studies involving more than 13,735 postmenopausal women were summarized and analyzed. In prospective studies, the effect of HRT on osteoporosis (OP) has been well documented. The effect of OP on mandibular bone density has also been examined in some observational studies. Few studies, however, have examined the effect of HRT directly on mandibular bone density or on dental outcomes. From these studies, the effect of HRT on costs of dental treatment and prophylaxis was estimated directly and indirectly. It was determined that HRT use was associated with reduction in adverse dental outcomes and the associated costs of dental care. Annualized excess cost in a cohort of 1,000 untreated women averaged $100,000. From this analysis, it is clear that postmenopausal women with OP who do not receive HRT have a greater incidence of adverse dental outcomes and higher dental care costs than those who do. PMID- 11066292 TI - Get big bang for your bucks with ESRD program. PMID- 11066293 TI - Improve scheduling of fistula patients to slash unnecessary costs, improve care for ESRD. AB - When you're dealing with a high-cost illness like ESRD, finding an easy source of six figure savings while enhancing access and quality is a significant accomplishment. But that's exactly what one hospital has done, using a simple strategy that virtually every organization with ESRD patients can adopt. PMID- 11066294 TI - Will early intervention keep schizophrenia at bay? AB - A growing body of research is focused on the prospect of preventive therapy for schizophrenia. Early intervention, if successful, could save millions of dollars in inpatient hospitalization, long-term individual and family psychotherapy, and vocational and psychosocial rehabilitation. PMID- 11066295 TI - Community-based DM approach slashes incidence of lead poisoning in children. AB - While lead poisoning among children is on the decline, it is still a serious health problem that affects an estimated 930,000 children between the ages of one and five. Baltimore health care providers have--with great success--turned to the tools of disease management to cut blood lead levels in children while simultaneously cutting overall costs. PMID- 11066296 TI - New guidelines prevent costly adverse drug reactions. AB - New guidelines on disease management of drug hypersensitivity have been released by the American College of Allergy, Asthma, & Immunology. The guidelines deal extensively with preventing adverse drug reactions, which represent a major source of unnecessary expense for hospitals and health plans. PMID- 11066297 TI - DM programs take different roads to CHF success. PMID- 11066298 TI - Boost outcomes by adding a clinical pharmacist to the disease management team. AB - Because of the importance of drug therapy in managing heart failure, Duke University Medical Center wanted to research the impact of adding a clinical pharmacist to its CHF disease management team. Their strategy--and the impressive results--could be adapted by any DM effort in which appropriate drug therapy plays a major. PMID- 11066299 TI - Humana buys into Web-based surveys to ID high-risk members. AB - Humana Inc., eager to identify high-risk members in a cost-effective way and improve patient outcomes, plans to administer more than 3 million health assessments over the Internet in the next two years. Here's how, including a copy of their online assessment report. PMID- 11066300 TI - What does it take to be a high-performance disease manager? AB - Organizations making the move toward formalized disease management must define the exact roles of DM team members and get the best-equipped people into those positions. This DM job description is a good. PMID- 11066301 TI - Clinton 2001 budget proposal damages hospitals. PMID- 11066302 TI - Revised CBO figures issued/BBRA gains for Medicare jeopardized. PMID- 11066303 TI - Medicare estimating: a multi-billion dollar guessing game that is hammering health care. PMID- 11066304 TI - Congressional outlook: health legislation. PMID- 11066305 TI - Medical records privacy--the winding (and bumpy!) road to implementation. PMID- 11066306 TI - HCFA rulings on DSH provide $5 billion windfall for northeast, others. PMID- 11066307 TI - Institute of Medicine study puts the spotlight on patient safety issues. PMID- 11066308 TI - Eight new "safe harbors" announced by Inspector General. PMID- 11066309 TI - Medicare can turn anyone into a crook. PMID- 11066310 TI - A tortoise at the starting line--more BBA relief? PMID- 11066311 TI - President Clinton raises the stakes in patient safety debate. PMID- 11066312 TI - Protecting patients' privacy while ensuring medical research. PMID- 11066313 TI - Campbell's antitrust exemption for docs gains momentum. PMID- 11066314 TI - Prompt pay to be resolved in coming weeks. PMID- 11066315 TI - Experts outline tips for navigating the post-acute terrain. PMID- 11066316 TI - Keeping up with changing managed care rules. PMID- 11066318 TI - Establishing realistic goals for disease management initiatives. PMID- 11066317 TI - A simple click of the mouse revolutionizes physician offices. PMID- 11066319 TI - Hospitalists: not for larger facilities only. PMID- 11066320 TI - Consider impact of health status when developing capitation rates. PMID- 11066321 TI - Consider the impact of MIS technology on your cap rate. PMID- 11066322 TI - Benchmark data help build your risk contracting strategy. PMID- 11066324 TI - Simple formula helps nail down your pharmacy PMPM costs. PMID- 11066323 TI - Data on specialty, PCP cap rates suggest wisdom of shared risk. PMID- 11066325 TI - Data provide ammunition to prepare for Medicaid contracting. PMID- 11066326 TI - 'Top 50' CPT codes help to predict overall costs. PMID- 11066327 TI - Spreadsheet shows importance of risk-adjusting your cap rate. PMID- 11066328 TI - Forms can help you evaluate feasibility of capitation rates. PMID- 11066329 TI - AMA database offers invaluable benchmark cap rates. PMID- 11066330 TI - Use these benchmark PMPMs to help steer capitation rates toward middle ground. PMID- 11066332 TI - Follow this blueprint for pricing your capitated contract. PMID- 11066331 TI - National survey offers snapshot of break-even PMPMs. PMID- 11066333 TI - Sample capitation rates provide PMPM benchmarks for senior, commercial plans. PMID- 11066334 TI - Survey provides wealth of capitation rate comparisons. PMID- 11066335 TI - Capitation survey provides invaluable capitation data. PMID- 11066336 TI - Use actuarial cost models to allocate global risk. PMID- 11066337 TI - Gauge the effect of co-pays on utilization, PMPM rates. PMID- 11066338 TI - Influence of capitation pushes bed days per 1,000 to record low levels. PMID- 11066339 TI - Refine medical management techniques to dramatically reduce utilization rates. PMID- 11066340 TI - Survey documents HMO and POS utilization rates, use of prevention measures. PMID- 11066341 TI - Chart your organization against benchmark bonanza. PMID- 11066342 TI - Wisconsin study compares managed care, fee-for-service Medicaid utilization rates. PMID- 11066343 TI - AMGA utilization data help providers anticipate senior market. PMID- 11066344 TI - Medicare, Medicaid study offers outpatient, hospital utilization data. PMID- 11066345 TI - Benchmark your costs against this data bonanza. PMID- 11066346 TI - Can capitation survive outside the urban jungle? PMID- 11066347 TI - Use patient self-assessments to identify high-risk patients, predict future health costs. PMID- 11066348 TI - Ensuring access becomes critical issue for managed care success. PMID- 11066349 TI - Beware risk contracts that skimp on behavioral health benefits. PMID- 11066351 TI - Fixed, variable cost analysis can dispel cost reduction myths, aid with decision making. PMID- 11066350 TI - Actionable data from claims drives integrated QI effort. PMID- 11066352 TI - Integrity panel focuses on individual data fields in quest to improve accuracy. PMID- 11066353 TI - Orthopedic benchmarks from top 100 hospitals show high volume means better care. PMID- 11066354 TI - Controlling water pollution in construction. PMID- 11066355 TI - Cleaning water storage tanks. PMID- 11066356 TI - Legionnaires disease and the Legionella risk assessment process. PMID- 11066357 TI - Control of Cryptosporidium in water systems using cartridge filtration. PMID- 11066358 TI - Case study of a silver/copper ionisation water treatment system in a district general hospital. PMID- 11066359 TI - Building using sustainable resources. PMID- 11066360 TI - Mental health centre. PMID- 11066361 TI - PFI--getting the best quality outcome. PMID- 11066362 TI - Reconstructing paradise: Canada's health care system, alternative medicine and the Charter of Rights. PMID- 11066363 TI - Wrongful life actions as a means of regulating use of genetic and reproductive technologies. PMID- 11066364 TI - Compensating family caregivers: an analysis of tax initiatives and pension schemes. PMID- 11066365 TI - A do not resuscitate order for an infant against parental wishes: a comment on the case of Child and Family Services of Central Manitoba v. R.L. and S.L.H. PMID- 11066366 TI - End-of-life decision making: rethinking the principles of fundamental justice in the context of emerging empirical data. PMID- 11066367 TI - Commercialization of genetic research and its impact on the communication of results. AB - Canada has recently seen significant commercial growth in biotechnology; at the same time we have witnessed a considerable reduction in public funding for research. One result is the development of partnerships between academic institutions and industry, which has had important effects on the relationships between researchers, companies, research subjects and society, particularly in the field of genetics. Commercialization of research creates obstacles to the diffusion of research results which is fundamental to the advancement of science. Several recent studies and cases, which are briefly reviewed here, have highlighted these problems. In this paper, the author examines clauses in research contracts in order to analyze and categorize the types of provisions these contracts may contain regarding publication and disclosure of research results. She then discusses the relationships between various actors in genetic research and the issues and conflicts that may arise. Finally, an examination of some recently developed policies in this area reveals the complex network of norms to which a researcher must adhere. The normative framework must take into account the interests of all the various actors, should apply to the broadest possible population, and its various parts must be consistent. Researchers must then be vigilant that they do not enter into contracts which conflict with their rights and obligations regarding publication and dissemination of results. PMID- 11066368 TI - Re-framing the discussion: commercial genetic testing in Canada. PMID- 11066369 TI - Reconstructing geneticization: a research manifesto. PMID- 11066370 TI - Patenting DNA and amino acid sequences--an Australian perspective. PMID- 11066371 TI - The mandatory reporting of child abuse under the Child Welfare Act. PMID- 11066373 TI - Safe and effective use of interpreters. PMID- 11066372 TI - Prehospital rounds. Pulmonary embolism. PMID- 11066374 TI - Into the funnel, down the tunnel. PMID- 11066375 TI - Oklahoma City remembers. PMID- 11066376 TI - A matter of time. Part I: Is EMS ready for domestic terrorism? PMID- 11066377 TI - The structure and role of DMAT teams. PMID- 11066378 TI - America's tragedy: pediatric trauma. AB - Resuscitation of the pediatric trauma patient involves immediate assessment of ABCs. Interventions are made immediately upon recognition of abnormalities during the primary survey. After initial assessment and management of life-threatening processes, consider the optimal destination for further care and resuscitation. Keep in mind pediatric considerations like anatomical differences and the need for pediatric-sized equipment. Avoid the common errors that occur when resuscitating a pediatric trauma patient. PMID- 11066379 TI - Pediatric cardiac emergencies. PMID- 11066380 TI - Airway management. PMID- 11066381 TI - Third World EMS. U.S. Peace Corp volunteer experiences EMS--Nicaraguan style. PMID- 11066382 TI - A guide to pediatric healthcare. PMID- 11066383 TI - Protect America's health care. PMID- 11066384 TI - Budget surpluses, or you should have seen the one that got away. PMID- 11066385 TI - No guarantees, only opportunities: an interview with Rodney J. Lisonbee. AB - Rodney J. Lisonbee, MAcc, CPA, is chief financial officer of the Urban South Region of Inter-mountain Health Care. IHC is a vertically integrated, not-for profit healthcare system providing service in Utah and Idaho. Operations of the Urban South Region include three hospitals, seven physician clinics, and a home healthcare agency. Lisonbee's entire career in health care has been with Intermountain. He joined the organization in 1985 as director of accounting for Intermountain's LDS Hospital in Salt Lake City, Utah and assumed the CFO position with the Urban South Region in 1988. PMID- 11066386 TI - IDSs reconsider the viability of risk contracting. AB - As financial success with risk contracting continues to elude many IDSs, some of these organizations have started questioning their overall mission as risk bearing entities. To determine whether to continue to assume risk, an IDS should consider its market environment, the commitment of its provider constituents to the organization, its core competencies, the likelihood of success with risk contracting, and alternatives. To determine its future course, the IDS should engage in a strategic-planning process that includes all IDS stakeholders. Such a process can help build consensus regarding the IDS's market characteristics and benefits of IDS membership; determine the IDS's appropriate roles, services, and functions; and evaluate resources required to pursue the desired strategy. The process should include planning for various possible future scenarios, and results should be communicated to all IDS provider constituents. PMID- 11066387 TI - Using scenario analysis to determine managed care strategy. AB - In today's volatile healthcare environment, traditional planning tools are inadequate to guide financial managers of provider organizations in developing managed care strategies. These tools often disregard the uncertainty surrounding market forces such as employee benefit structure, the future of Medicare managed care, and the impact of consumer behavior. Scenario analysis overcomes this limitation by acknowledging the uncertain healthcare environment and articulating a set of plausible alternative futures, thus supplying financial executives with the perspective to craft strategies that can improve the market position of their organizations. By being alert for trigger points that might signal the rise of a specific scenario, financial managers can increase their preparedness for changes in market forces. PMID- 11066388 TI - Stacked penalties raise stakes in fraud and abuse prosecutions. AB - Although providers try to comply with the laws regulating the provision of healthcare services to Medicare patients, there are many circumstances under which violations could be committed unwittingly, making the provider liable for civil or criminal penalties. If a provider becomes aware that a violation of the Federal antikickback statute, the Stark II law, or the False Claims Act may have occurred, it should act quickly to determine if, in fact, a violation actually did occur and take corrective action, or document why the particular activity does not constitute a violation. In some cases, a violation of the False Claims Act can be linked with a violation of one of the other laws. When violations are "stacked" in this manner, provider liability increases. PMID- 11066389 TI - Broader range of skills distinguishes successful CFOs. AB - In recent years, healthcare CFOs have seen their role expand significantly beyond traditional financial duties. A series of trended surveys on CFO roles and responsibilities reveals that today's healthcare CFO requires a broad new range of traits and skills in the areas of leadership, operations, and healthcare strategy. CFOs regard strategic thinking and the ability to communicate clearly as the most important of their essential leadership traits and skills, respectively. Among operational and strategic skills, CFOs most often cite the importance of being able to improve organizational performance and benchmark. Healthcare CFOs can enhance their chances of success by focusing self-improvement efforts on five key areas: implementing the organization's vision; developing tactics that stimulate change; enhancing communication skills; focusing on managing and leading; and strengthening relationships. PMID- 11066390 TI - Internal investigations: a proactive approach to compliance. AB - Healthcare financial managers increasingly are recognizing that internal investigations can uncover and help correct improper activities before they attract government attention, as well as reveal valuable facts about any questionable activities that may be the target of a government inquiry. An organization that exhibits a high level of responsibility by undertaking a thorough internal investigation of possible improper activities may reduce or eliminate civil or criminal sanctions the government can impose if the organization is found guilty of improprieties. Furthermore, an internal investigation can preserve the organization's integrity and reaffirm its commitment to high ethical standards and sound business practices. PMID- 11066391 TI - The evolution of hospitalist programs. AB - In just a few years, hospitalist programs have evolved to a third generation, affording key participants an opportunity to reduce inpatient healthcare costs. Because the method of inpatient payment determines the economic winners and losers in the program, the key participants will benefit from the implementation of a program that includes shared incentives. Even though reduced payments for inpatient services might be offset by reduced variable costs for hospitals and increased office revenues for primary care physicians, a hospitalist program is more likely to achieve sustained success if it is structured so that financial rewards are distributed fairly among the participants. PMID- 11066392 TI - More BBA payment relief is on the horizon. PMID- 11066393 TI - ASPs can assist compliance efforts. PMID- 11066394 TI - Durable power of attorney: a powerful planning tool. PMID- 11066395 TI - New information technology opportunities challenge CFOs. PMID- 11066396 TI - Data trends. Tracking coding practices under APCs. PMID- 11066397 TI - Eleventh Amendment limits upon the private enforceability of state hospitals' federal legal obligations. PMID- 11066398 TI - Peer review protection statute blocks aggrieved physician. PMID- 11066399 TI - Wireless POCT data transmission. AB - Despite the many advantages of POCT, the biggest obstacle for the lab often becomes documentation and billing of tests performed at remote locations. To solve this problem, this lab took advantage of a wireless radiofrequency network that had been installed for the pharmacy to transmit and receive patient histories. They decided to use this same system to communicate between the analyzers and the LIS. PMID- 11066400 TI - Getting to the point: integrating critical care tests in the patient care setting. AB - Implementing testing services in the patient care setting offers the lab manager many challenges. At this hospital system, an innovative data management solution addressed the problems of a simple user interface, automatic creation of test orders and accession numbers, remote review of test results by the central laboratory, and an interface to the LIS and HIS for complete data capture. PMID- 11066401 TI - When and how to screen for liver disease. PMID- 11066402 TI - Big suppliers strike Internet deal. PMID- 11066403 TI - HCFA hit on settlements. Alleged provider 'sweetheart deals' seen as bad precedent. AB - Call it selective prosecution. Last week's disclosure of several alleged "sweetheart deals" involving the settlement of Medicare billing disputes has HCFA officials concerned that other healthcare providers could start demanding more favorable treatment. Former HCFA Administrator Bruce Vladeck (left) was among those testifying on those cases last week in Washington. Meanwhile, a look at data on federal fraud investigations shows big geographic disparities in those probes. See story on p. 36. PMID- 11066404 TI - More gloom from hospitals. PMID- 11066405 TI - Managed care, providers in rift. AAHP's ad campaign on medical errors causes a round of industry finger-pointing. PMID- 11066406 TI - Playing hospital in Phoenix. Infrastructure is real; patients aren't. PMID- 11066408 TI - Healthcare fraud bug is opportunistic. PMID- 11066407 TI - The geography of fraud. A concentration of cases in the northeast, Fla. PMID- 11066409 TI - CHI posts first-time loss. Catholic Health Initiatives. PMID- 11066410 TI - Doc antitrust exemption advances. PMID- 11066411 TI - Solutions in the mist. PMID- 11066412 TI - Installations help fight medical errors. PMID- 11066413 TI - A vocal advocate. Interview by Ron Shinkman. PMID- 11066414 TI - Not-for-profits hope to sway investors. PMID- 11066415 TI - AMA steps up the pressure. PMID- 11066416 TI - Still not enough. PMID- 11066417 TI - McKesson HBOC reorganizes IT unit. PMID- 11066418 TI - Just call it gpo.com. Internet deals change the supply biz. PMID- 11066419 TI - Ala. system to sell HMO. PMID- 11066420 TI - N.M. CEO exits amid concern over projects. PMID- 11066421 TI - Church vs. state. PMID- 11066422 TI - South Florida gets into quality. PMID- 11066423 TI - Two provider-owned HMOs probed. PMID- 11066424 TI - Elections break impasse--for now. PMID- 11066425 TI - New ammo. PMID- 11066426 TI - Unnecessarily complex. PMID- 11066427 TI - Internet dominates providers' line of sight. PMID- 11066428 TI - Time crunch is expected in meeting APC deadline. PMID- 11066429 TI - Cost standard can help APC analysis. PMID- 11066430 TI - AAA stent-grafts less invasive. PMID- 11066431 TI - Benchmarking OR turnover times. PMID- 11066432 TI - Improving patient flow wins award. PMID- 11066433 TI - So who wants to be a supervisor? PMID- 11066434 TI - Implant sets: time for a change? PMID- 11066435 TI - Managers taking on RN training. PMID- 11066436 TI - Materials management. New ventures crowd e-commerce. PMID- 11066437 TI - Getting your charge description master into shape. PMID- 11066438 TI - HCPCS/revenue code chart. PMID- 11066439 TI - Strategic communications build academic health center's brand. PMID- 11066440 TI - Dining out at your Web site. PMID- 11066441 TI - Dropping a Medicare HMO without losing face. PMID- 11066442 TI - Sentara uses merger consolidation to extend its brand awareness. PMID- 11066443 TI - Marketing program opens Ochsner's doors, but first it hits the employees. PMID- 11066444 TI - Community cardiac response project enhances system's image. PMID- 11066445 TI - Rich content drives good health care Web sites. PMID- 11066446 TI - Utilization management remains elusive goal. PMID- 11066447 TI - MR can help save victims of stroke, but challenges remain. PMID- 11066448 TI - How to encrypt medical data against intrusion. PMID- 11066449 TI - Acute promyelocytic leukemia (AML-M3)--Part 2: Molecular defect, DNA diagnosis, and proposed models of leukemogenesis and differentiation therapy. AB - OBJECTIVES: To identify the chromosomal translocation common in M3 and discuss its diagnostic use to: Compare acute leukemia with chronic leukemia and other forms of cancer. Describe the molecular defect including the fusion gene and fusion protein produced from the translocation. Discuss the proposed mechanism of leukemogenesis in M3. Discuss the proposed mechanism of differentiation induction stimulated by ATRA therapy. Present the future direction of this and other forms of therapy. DATA SOURCES: Current literature. DATA SYNTHESIS: Acute promyelocytic leukemia (AML-M3) is a form of acute leukemia that presents with a less dramatic leukocytosis, anemia, and thrombocytopenia than other acute leukemias. However, AML-M3 has a lower first remission rate and a higher morbidity and mortality rate than most of the other acute leukemias when treated with conventional chemotherapy. AML-M3 frequently stimulates a serious concomitant coagulation disorder, disseminated intravascular coagulation, which is a major contributor to the high mortality rate. This and other devastating sequela of M3 have prompted clinicians and investigators to develop methods of improving diagnosis and therapy. In 1977 the method of diagnosis confirmation was improved by the identification of a consistent chromosomal translocation involving the long arms of chromosomes 15 and 17. Identification of the specific molecular lesion that produced the t(15;17) translocation occurred in 1990 and was shown to involve the retinoic acid receptor alpha gene (RAR alpha). Because the RAR alpha gene is mutated in all AML-M3 patients studied so far and because it is often the only mutation identified, several proposed mechanisms of leukemogenesis have evolved. From these discoveries a novel approach to cancer treatment focusing on differentiation therapy instead of traditional chemotherapy emerged. All-trans retinoic acid (ATRA) has been shown to stimulate differentiation of promyelocytes from the malignant clone and has become an important element in the treatment of patients with AML-M3. CONCLUSION: Since the discovery of the t(15;17) translocation, the identification of the fusion gene containing the retinoic acid receptor alpha, and the success of ATRA as a form of differentiation therapy, the diagnosis and treatment of AML-M3 has dramatically improved. In addition, AML-M3 has become a model system used to study the mechanisms that produce uncontrolled growth and lack of differentiation in leukemic cells (leukemogenesis) and the mechanisms of therapeutic reversal of this block in differentiation (differentiation therapy). PMID- 11066450 TI - Immunophenotyping leukemias: the new force in hematology. AB - OBJECTIVE: To review basic concepts of immunophenotyping leukemias and introduce clinical laboratory scientists (CLSs) to the technological changes utilized in this laboratory methodology. DESIGN: Literature review. DATA SYNTHESIS: Immunophenotyping in the clinical laboratory is emerging as an advantageous way to separate and classify leukemic malignancies. Immunophenotyping involves the use of flow cytometers and immunofluorescence in order to achieve great sensitivity and specificity for malignant cells. A basic understanding of components of the flow cytometer and how it works is necessary to understand immunophenotyping. Monoclonal antibodies specific to the malignant cells of question play an essential part in this technique. Various fluorescent dyes and cell panels also must be incorporated into the system. Analysis is done and statistics are plotted on dot plots that can be read by the CLS to give helpful insight into the etiology of disease process. Immunophenotyping is a very powerful tool that has the ability to revolutionize the clinical laboratory setting. The CLS working in hematology must become aware of and comfortable with this methodology. PMID- 11066451 TI - CLS education at a new crossroad. AB - OBJECTIVE: To provide an overview of the history of CLS education at selected points in time; to describe issues facing CLS education today; and to discuss possible directions for CLS education in the future. PMID- 11066452 TI - The 2000 federal legislative agenda for healthcare: real reform or political rhetoric? PMID- 11066453 TI - Updating the immunology curriculum in clinical laboratory science. AB - OBJECTIVE: To determine essential content areas of immunology/serology courses at the clinical laboratory technician (CLT) and clinical laboratory scientist (CLS) levels. DESIGN: A questionnaire was designed which listed all major topics in immunology and serology. Participants were asked to place a check beside each topic covered. For an additional list of serological and immunological laboratory testing, participants were asked to indicate if each test was performed in either the didactic or clinical setting, or not performed at all. SETTING: A national survey of 593 NAACLS approved CLT and CLS programs was conducted by mail under the auspices of ASCLS. PARTICIPANTS: Responses were obtained from 158 programs. Respondents from all across the United States included 60 CLT programs, 48 hospital-based CLS programs, 45 university-based CLS programs, and 5 university based combined CLT and CLS programs. MAIN OUTCOME MEASURES: The survey was designed to enumerate major topics included in immunology and serology courses by a majority of participants at two distinct educational levels, CLT and CLS. Laboratory testing routinely performed in student laboratories as well as in the clinical setting was also determined for these two levels of practitioners. RESULTS: Certain key topics were common to most immunology and serology courses. There were some notable differences in the depth of courses at the CLT and CLS levels. Laboratory testing associated with these courses also differed at the two levels. Testing requiring more detailed interpretation, such as antinuclear antibody patterns (ANAs), was mainly performed by CLS students only. CONCLUSION: There are certain key topics as well as specific laboratory tests that should be included in immunology/serology courses at each of the two different educational levels to best prepare students for the workplace. Educators can use this information as a guide to plan a curriculum for such courses. PMID- 11066454 TI - Determining clinical laboratory science curriculum for the 21st century. AB - OBJECTIVE: To conduct a study to show possible differences in clinical laboratory science (CLS) education in relation to knowledge and skill levels deemed most important to job performance success of entry bench level CLS practitioners as determined by laboratory supervisors. Information gained from the study may indicate areas of program curriculum needing revision, or the incorporation of subject areas not presently offered. DESIGN: Survey. PARTICIPANTS: CLS educators from 100 different hospital-based and university-based CLS programs, and medical laboratory departmental supervisors from 209 different hospital laboratories. OUTCOME MEASURES: An analysis of the data from the survey consisted of individual item percentages generated by both surveys and a comparison of tasks deemed highly important by supervisors with class time estimates devoted to those tasks. RESULTS: The study indicated differences between what supervisors viewed as important knowledge and skills of entry bench level CLSs and the amount of class time devoted to those subjects by CLS educators. CONCLUSIONS: To ensure continuing professional credibility, additional study will be needed regarding the education and practice of CLSs as automation, emerging technologies, and laboratory restructuring will continue to change the laboratory environment. PMID- 11066455 TI - Acute promyelocytic leukemia (AML-M3)--Part 1: Pathophysiology, clinical diagnosis, and differentiation therapy. AB - OBJECTIVES: The objectives of this review paper are to: Compare acute leukemia with chronic leukemia and other forms of cancer. Review the pathophysiology and discuss the clinical and diagnostic features of M3. Describe the variant of M3 (M3m or M3v). Compare conventional chemotherapy with all-trans retinoic acid differentiation therapy (ATRA) in the treatment of M3. Discuss the future direction of differentiation therapy. DATA SOURCES: Current literature. DATA SYNTHESIS: Until the late 1970s, the methods of diagnosis and treatment of AML-M3 were similar to the other forms of acute myelocytic leukemia. One notable difference between M3 and other acute myelocytic leukemias involved the common occurrence of life-threatening consumptive coagulopathies in M3 patients that dramatically affected patient outcomes. In 1977 the method of diagnosis confirmation was altered by the identification of a consistent chromosomal translocation involving the long arms of chromosomes 15 and 17. Reports in the 1970s and 1980s indicated that certain types of active metabolites of vitamin A, collectively termed retinoids, could induce differentiation in a variety of cancer derived cell lines, in vitro. It was shown in the early 1980s that 13-cis retinoic acid (13cRA) could stimulate differentiation in bone marrow cells collected from patients with various acute leukemias, including M3. The first clinical trial of a retinoid, given as a remission induction therapeutic regimen, involved all-trans retinoic acid (ATRA) administered to M3 patients in 1988. Since then, ATRA therapy has been shown to reduce tumor burden by stimulating differentiation of the leukemic cells, induce long-term clinical remission when administered in conjunction with chemotherapy, and lower the incidence of consumptive coagulopathies in patients with AML-M3. CONCLUSION: The diagnosis and treatment of AML-M3 has improved dramatically in the past decade, which has greatly enhanced the prognosis of patients with this disease. First remission rates have increased to greater than 85% world wide, the incidence of disseminated intravascular coagulation (DIC) has declined dramatically, and 60% to 70% of patients with AML-M3 have achieved long term survival and are potentially cured. PMID- 11066456 TI - Health education in Africa: 1975-2000. PMID- 11066457 TI - Does locus of control moderate the effects of tailored health education materials? AB - Research in health communication has shown that individually tailored health education materials are more effective than traditional or generic materials in producing changes in health-related behaviors. However, tailored materials have not been equally effective for all individuals. Because locus of control affects behavioral outcomes in other self-change interventions, its effect on individuals' responses to tailored messages is of particular interest. The present study examined differences in cognitive responses to tailored and non tailored weight loss materials among 198 overweight individuals. Weight locus of control significantly interacted with study group (who received either tailored or non-tailored materials), suggesting that externals may respond to tailored health education materials with counter-arguments. Implications for the development and application of tailored health communication materials are discussed. PMID- 11066458 TI - Modeling tanning salon behavioral tendencies using appearance motivation, self monitoring and the theory of planned behavior. AB - The constructs of appearance motivation and self-monitoring were added to the Theory of Planned Behavior in the prediction of tanning salon use in young people. The variables of the Theory of Planned Behavior proved effective at predicting tanning salon behavioral intentions and tendencies. Intentions and perceived behavioral control predicted tanning salon behavioral tendencies, while attitudes, subjective norms and perceived behavioral control predicted tanning salon behavioral intentions. Appearance motivation did not show any direct or interaction effects in the prediction of tanning salon behavioral intentions. It did, however, prove superior to health orientation in the prediction of tanning salon attitudes. Self-monitoring interacted with subjective norms in the prediction of tanning salon intentions, with high self-monitors showing stronger subjective norm-intention relationships than low self-monitors. These results imply that appearance-related interventions could prove efficacious in reducing young people's tanning salon behavioral tendencies. Furthermore, it may be important to consider individual's self-monitoring status when targeting skin cancer prevention information to young people. PMID- 11066459 TI - Psychosocial predictors of smoking among secondary school students in Henan, China. AB - The objective of this study was to measure the risk factors associated with tobacco use among secondary school students in Henan, China. Self-reported questionnaires were administered to four secondary schools; 3519 students were studied including 1799 boys and 1720 girls aged 10-19. Demographic, behavioral, attitudinal/belief, knowledge and interpersonal variables were investigated. Overall, 15.1% of boys and 1.4% girls reported smoking at least occasionally. Smoking onset is most prevalent from the ages of 10-14. The smoking rate increased with age. The likelihood of tobacco use was significantly higher among those having peers, teachers or mother who smoked. Positive smoking-related attitudes among students had a significant association with their smoking status. The results suggest that effective smoking prevention interventions need to be comprehensive and implemented in the early teen years. PMID- 11066460 TI - Subtypes of precontemplating smokers defined by different long-term plans to change their smoking behavior. AB - Many smokers are not motivated to quit smoking. In the Stages of Change model these smokers are called precontemplators. When developing interventions designed to motivate these smokers to quit, it is of importance to know whether this group is homogeneous or not. In the present study, different groups of precontemplators were distinguished according to their long-term quitting smoking plan: 861 precontemplators were asked to indicate the one plan that best fitted their own plans with regard to their smoking behavior: (1) planning to never quit and not planning to cut down (n = 194), (2) planning to never quit but planning to cut down (n = 186), (3) planning to quit somewhere in the future but not within the next 5 years (n = 290), (4) planning to quit within the next 5 years (n = 136) and (5) planning to quit within the next year but not within the next 6 months (n = 54). These groups of smokers were compared on several variables cross sectionally and longitudinally. The results indicate that the psychological factors that will have to be targeted in smoking cessation interventions in efforts to motivate smokers to quit could be assessed reliably in precontemplators. Furthermore, precontemplators with different quitting plans differed on several cognitive variables and the quitting plans at pre-test were predictive of quitting activity after 7 months. Precontemplators who received self-help smoking cessation materials made forward changes in quitting plans and these changes seemed to follow a certain order. Forward changes in plans were differentially related to positive outcome expectations, to self-efficacy expectations depending on the quitting plan and not to changes in negative outcomes. The present study is one step in mapping the psychology of low motivation to change behavior. PMID- 11066461 TI - Stages of change in two modes of health-enhancing physical activity: methodological aspects and promotional implications. AB - Measurement scales for stages of change were developed and the stages were assessed in two specific modes of Health-Enhancing Physical Activity (HEPA) in a cross-sectional survey (N = 1516); representative samples were selected from three age groups, i.e. from three phases of adult life. Outdoor Aerobic Exercise (OAE) was used as an example of fitness activity; Everyday Commuting Activity (ECA) was selected to represent lifestyle physical activity. Scales used by the Prochaska team were modified for this study, and the stages of Precontemplation and Preparation were each divided into two new stages. Consistency of the stage measurement was moderate for OAE and good for ECA. As regards content validity, consistent associations were found between stage scores and contextual variables for both behaviors. The results show that, at a given time, a person can be in different stages in different modes of HEPA. Therefore, the behavior of interest must be specified before accurate information on the stages of change in a population can be obtained. The results also indicate the importance of contextual factors in HEPA promotion. PMID- 11066462 TI - Tailoring dietary feedback to reduce fat intake: an intervention at the family level. AB - In this study, we wished to investigate whether the use of tailored nutrition education letters addressed to each family member simultaneously at home could serve as a valuable strategy for nutrition education. Family quartets (both parents and two adolescents, all healthy individuals) were chosen to be the units of intervention. The first aim of our study was to investigate the impact of tailored versus standardized nutrition education on fat intake and on psychosocial determinants of fat intake in families, using a randomized dietary feedback study. Our second aim was to study the differential effect of the tailored nutrition education on different family members. Analyses were conducted among 18 experimental families (n = 72) and 17 control families (n = 68). The tailored intervention was more effective than the nontailored intervention in reducing total and saturated fat intake when all the family members were included (F = 4.0, P < 0.05 and F = 5.9, P < 0.05). However, follow-up analyses revealed that only mothers benefit from the tailored intervention (F = 6.4, P < 0.05 and F = 10.2, P < 0.005). For fathers and adolescents, both interventions resulted in a significant decrease in fat scores. Furthermore, tailored feedback resulted in stronger awareness of personal fat intake and awareness of fat intake of family members. Tailored advice has the potential to communicate the personal need to change. As differences in fat reduction between family members receiving general or tailored nutrition education letters were smaller than expected, future research will have to prove that family-based tailored interventions are more effective than standardized interventions and interventions focusing on a single person. It also needs to be clarified why mothers in particular benefit from tailored feedback. PMID- 11066463 TI - A National Institute for Occupational Safety and Health Alert sent to hospitals and the intentions of hospital decision makers to advocate for latex allergy control measures. AB - This study evaluated a National Institute for Occupational Safety and Health (NIOSH), Centers for Disease Control and Prevention, Alert concerning the risk and prevention of latex allergy among health care workers. It has been estimated that 8-12% of health care workers are sensitized to latex. NIOSH Alerts are publications that are intended to educate stakeholders about risks in the workplace; this Alert contained four recommendations for administrative control measures that hospital decision makers could adopt to reduce the risk of latex allergy to employees. The Alert was mailed to a random selection of Directors of Infection Control and Directors of Nursing in hospitals in the US. A random sample of these targeted recipients and a control group were surveyed by telephone (N = 298). Although nearly all of the respondents were concerned about latex allergy (96%), those reporting having seen the Alert were significantly more likely to report an intention to advocate for one or more of the control measures. PMID- 11066464 TI - Process measures in an antenatal smoking cessation trial: another part of the picture. AB - Data on provider and patient compliance can be crucial in understanding the degree of a health education program's effectiveness, as well as in identifying areas where the program requires modification. However, such data are rarely systematically reported in randomized trials. This report assesses the degree to which doctors and midwives complied with intervention protocols in a hospital antenatal smoking cessation trial, and also examines the program's acceptability to patients. Provider compliance was assessed principally via consultation audiotapes and provider-completed checklists. The audiotape analysis identified substantial compliance problems. For example, in relation to six specific smoking related pregnancy risks, the proportions of Experimental Women informed about each individual risk ranged from 26 to 38% and the proportions receiving counselling items ranged from 52 to 79%. Doctors only informed a minority of Experimental Women of the increased risk of Sudden Infant Death Syndrome (28%) and of the presence of toxic chemicals in tobacco (21%). Comparison of compliance data from audiotapes and provider checklists revealed there was no significant agreement in three of four cases tested. Experimental Patients completed questionnaires to assess recall of smoking advice and to rate 12 program features. Of specific Experimental Program elements, the videotape (85%) received the highest level of positive patient ratings and the lottery (42%) the lowest. The process evaluation indicated that the Experimental Program needed some modification to increase its suitability for routine application. The findings also support the value of including an objective measure of provider compliance. PMID- 11066465 TI - Factor analysis of the Condom Use Self-Efficacy Scale among multicultural college students. AB - The Condom Use Self-Efficacy Scale (CUSES) was administered to 447 multicultural college students. The sample consisted of 63.5% Hispanic/Latino, 17.1% African American, 13.7% Caucasian, 4.1% other and 1.6% Asian students. The obtained scores were subjected to a principal components factor analysis with a Varimax rotation. An item designation criteria was used and three distinct factors were extracted: (1) 'Appropriation', (2) 'Sexually Transmitted Diseases' and (3) 'Partners' Disapproval'. Comparisons to the only other published factor analysis of the CUSES are made. Implications for future research using the CUSES to design AIDS education curricula for multicultural college students are discussed. PMID- 11066466 TI - A pilot study to establish a randomized trial methodology to test the efficacy of a behavioural intervention. AB - How can pregnant women be helped to stop smoking? This was a pilot study of midwife home-based motivational interviewing. Clients were 100 consecutive self reported smokers booking at clinics in Glasgow from March to May 1997. Smoking guidance is routinely given at booking. In addition, intervention clients received a median of four home-based motivational interviewing sessions from one specially trained midwife. All sessions (n = 171) were audio-taped and interviews (n = 49) from 13 randomly selected clients were transcribed for content analysis. Three 'experts' assessed intervention quality using a recognized rating scale. Cotinine measurement on routine blood samples confirmed self-reported smoking change from late pregnancy telephone interview. Postnatal telephone questionnaire measured client satisfaction. Focus groups of routine midwives explored acceptability, problems and disruption of normal care. Fisher exact, chi 2 and Mann-Whitney tests compared enrolment characteristics. Two-sample t-tests assessed outcome between groups. Motivational interviewing was satisfactory in more than 75% of transcribed interviews. In this pilot study, self-reported smoking at booking (100 of 100 available) corroborated by cotinine (93 of 100) compared with late pregnancy self-reports (intervention 47 of 48; control 49 of 49) and cotinine (intervention 46 of 48; control 47 of 49) showed no significant difference between groups. Tools have been developed to answer the question: 'Can proactive opportunistic home-based motivational interviewing help pregnant smokers reduce their habit?'. PMID- 11066467 TI - Response to Kreuter and Skinner. PMID- 11066468 TI - Advocacy: an IUHPE priority. PMID- 11066469 TI - Clinical commentary: the law of unintended ethics. AB - Criticizes Ronald Green's parental-based approach to genetic testing and counseling of children for its failure to address the clinical realities of genetic practice. PMID- 11066470 TI - Legal and ethical commentary: the dangers of reading duty too broadly. AB - Criticizes Ronald Green's proposal for its failure to consider the disastrous consequences his proposals would have were they to become legal mandates. PMID- 11066471 TI - Substituted judgment in medical practice: evidentiary standards on a sliding scale. AB - Proposes an approach to the substituted judgment standard that reflects the reality of clinical practice and circumvents shortcomings in the current legal and bioethical alternatives. PMID- 11066472 TI - Medicaid, managed care, and America's health safety net. AB - Argues that managed care, by design, is poorly suited as a model to solve the growing problem of financing the health care needs of Medicaid populations. PMID- 11066473 TI - Creutzfeldt-Jakob disease: the problem of recipient notification. AB - Considers the legal and ethical obligations of blood suppliers to warn recipients of blood products about possible contamination of the blood supply with Creutzfeldt-Jakob disease, a rare but fatal neurological disease. PMID- 11066474 TI - Bioethics, public health, and firearm-related violence: missing links between bioethics and public health. AB - Examines assumptions underlying current themes in U.S. bioethics, and criticizes bioethicists for their failure to focus on public health, preventive medicine, and social medicine. PMID- 11066475 TI - Acetaminophen (Tylenol): Johnson & Johnson and consumer safety. AB - Considers, against backdrop of Johnson & Johnson's handling of the Tylenol poisonings of the 1980s, Johnson & Johnson's response to product safety concerns raised about acetaminophen by studies suggesting links to hepatotoxicity and end stage renal disease. PMID- 11066476 TI - Parental autonomy and the obligation not to harm one's child genetically. AB - Examines the potential use of genetic information by parents in ways that offer no preventive or curative effects and that may in fact inflict harm on children. PMID- 11066477 TI - Contracts: HMO arbitration agreements. PMID- 11066478 TI - Antitrust: appellate court to hear suit on antitrust implications of MCO contracts. PMID- 11066479 TI - Contracts: state extension-of-benefits law not preempted by ERISA. PMID- 11066480 TI - Evidence: appellate court dismisses expert testimony under Daubert standard. PMID- 11066481 TI - Legislative activity: funding reform and medical education. PMID- 11066482 TI - Medicaid & Medicare: Ninth Circuit overrules health services on medical reimbursement rates. PMID- 11066483 TI - Pharmaceuticals: FDA labeling time line. PMID- 11066484 TI - Products liability: Supreme Court denies federal preemption claims under MDA. PMID- 11066485 TI - Commentary: quality, costs, privacy and electronic medical data. AB - While agreeing that proper confidentiality measures are necessary when handling patient records, author discusses finding a balance between privacy and the benefits of using patient data to improve quality and to control costs. PMID- 11066486 TI - Medical record confidentiality law, scientific research, and data collection in the information age. AB - Author reviews U.S. federal and state laws on privacy and confidentiality on access and disclosure of health records governing government and private sector data bases. He also examines legislative proposals and recommendations for privacy and confidentiality now before the U.S. Congress. PMID- 11066487 TI - Privacy and confidentiality practices for research with health information in Canada. AB - Author outlines structurally the Canadian statutes and policies governing access to health information for research purposes at the federal level (Statistics Canada) and in three selected provinces. PMID- 11066488 TI - Decisionally impaired persons in research: refining the proposed refinements. AB - Amid the ethical debate about consent to research involving cognitively impaired subjects, author cautions against creating barriers to acceptable research and provides criteria by which guidelines should be developed. PMID- 11066489 TI - Institutional efforts to promote advance care planning in nursing homes: challenges and opportunities. AB - In an empirical study of Connecticut-area for-profit and nonprofit nursing homes, authors examine use of institution-specific advance care planning forms among nursing home residents. PMID- 11066490 TI - Tort liability for managed care: the weakening of ERISA's protective shield. AB - In light of recent developments in the law of ERISA preemption, author assesses the extent to which litigants are successfully imposing tort liability on HMOs and MCOs. PMID- 11066491 TI - Serious illness and private health coverage: a unique problem calling for unique solutions. AB - Authors examine the experience of two nonelderly adult populations in Indiana and their difficulties in obtaining and retaining health insurance once diagnosed with a serious chronic or catastrophic disease. PMID- 11066492 TI - Eliminating conflicts of interest in managed care organizations through disclosure and consent. AB - Author argues that, through disclosure of conflicts of interest and patient consent to these conflicts, MCOs can mitigate or even eliminate many conflicts of interest that arise with incentive plans or plan constraints. PMID- 11066493 TI - Commentary: a physician's perspective on conflicts of interest. AB - Author argues that, in clinical practice, disclosure of managed care incentives and other cost-constraint measures does not obviate physicians' primary duty to their patients. PMID- 11066494 TI - Religious exemptions to the immunization statutes: balancing public health and religious freedom. AB - Given the AAP's 1997 position statement on religious exemptions to medical care, authors consider whether failure to immunize a child is medical neglect. Although acknowledging that it is, they argue that parental decisions not to vaccinate on the basis of religious beliefs should be permitted. PMID- 11066495 TI - Managed care: gag clauses and doctor-patient communication: state responses. PMID- 11066496 TI - Fraud & abuse: DOJ and Medicare and Medicaid model compliance programs. PMID- 11066497 TI - Evidence: Oregon court excludes expert testimony in breast implant litigation. PMID- 11066498 TI - Licensing/disciplinary actions: Illinois Supreme Court upholds state restrictions on medical solicitation. PMID- 11066499 TI - Malpractice: district court holding affects ERISA preemption shield for HMO malpractice claims. PMID- 11066500 TI - Managed care: Eighth Circuit declares HMOs have fiduciary duty to disclose incentive structure. PMID- 11066501 TI - Medicare & Medicaid: Ninth Circuit decides that Medicare Act does not preempt wrongful death claim. PMID- 11066502 TI - Introduction: medical record confidentiality and data collection. PMID- 11066503 TI - Medical record confidentiality and data collection: current dilemmas. AB - Author discusses current data collection and data disclosure practices that threaten medical record confidentiality. She raises concerns about the reidentification of ostensibly deidentified patient information and discusses measures for protecting confidentiality in a research setting. PMID- 11066504 TI - Weaving technology and policy together to maintain confidentiality. AB - Author demonstrates that removing all explicit identifiers from medical data does not guarantee medical record confidentiality. She examines three new software systems that do help maintain anonymity, but warns that systems' limitations demand complementary policies. PMID- 11066505 TI - Medical malpractice implications of PSA testing for early detection of prostate cancer. AB - Prostate cancer screening is controversial. We conducted a national survey of primary care physicians and urologists to examine their use of prostate-specific antigen testing and their views on medical liability issues involved. PMID- 11066506 TI - Cancer genetic susceptibility testing: ethical and policy implications for future research and clinical practice. Cancer Genetic Studies Consortium, National Institutes of Health. AB - Authors examine the ethical and health policy implications in the Cancer Genetic Studies Consortium projects, which attempt to collect data on the clinical benefits and harms of cancer genetic testing. They suggest that more data are needed on the long-term physical and psychosocial effects of testing and that further examination is needed of the ethical issues raised by testing. PMID- 11066507 TI - Commentary: risks and benefits, testing and screening, cancer, genes and dollars. AB - Author argues that the current, restrictive policy for genetic screening for cancer risk is appropriate but that diagnostic testing decisions should not be so narrowly regulated. PMID- 11066508 TI - The Genetic Privacy Act: an analysis of privacy and research concerns. AB - Author argues that the Genetic Privacy Act fails to protect nucleic acid-based information as it relates to individual privacy, yet overburdens medical and scientific research with vague consent standards. PMID- 11066509 TI - Procreation by cloning: crafting anticipatory guidelines. AB - Author argues in favor of a nonlegislative approach to cloning that distinguishes among types of cloning and recognizes the conditions under which cloning would likely be sought. PMID- 11066510 TI - Professional self-regulation and shared-risk programs for in vitro fertilization. AB - Authors argue that risk-of-failure insurance for IVF, a form of contingent fee, may be ethically offered to patients if there is a full explanation of program details and advantages and disadvantages for patients. PMID- 11066511 TI - Money-back guarantees for IVF: an ethical critique. AB - Author contends that the moral arguments supporting money-back guarantees on infertility services are flawed because such services damage the physician patient relationship and threaten the meaning of parenthood. PMID- 11066512 TI - Bioethics and international human rights. AB - Noting how the spread of medical technology is creating clashes with traditional values and within cultures, the author addresses the clash between Western rights based incentives, as used by the United Nations to guarantee respect for life and dignity, and communitarian traditions. He proposes a mean between wholesale cultural relativism and international absolutism. PMID- 11066513 TI - Disability & ADA: Sixth Circuit affirms congressional intent of Title III. PMID- 11066514 TI - Legislative activity: HIPAA and recommendations to protect individual privacy. PMID- 11066515 TI - Antitrust: Fifth Circuit upholds dumping of hospital from network contract. PMID- 11066516 TI - Arbitration: California court clarifies arbitration clauses for CCLRA violations. PMID- 11066517 TI - Contracts: Sixth Circuit interprets COB clause to hold ERISA insurer liable. PMID- 11066518 TI - Employment: Sixth Circuit affirms spousal notification of COBRA rights. PMID- 11066519 TI - Employment: Ninth Circuit finds charge nurses not supervisors. PMID- 11066520 TI - Insurance: Eleventh Circuit interprets MSP statute definition of "group health plan". PMID- 11066521 TI - Licensing/disciplinary actions: Arizona court holds physician-reviewer accountable for precertification denial. PMID- 11066522 TI - Medicare & Medicaid: New York court denies nonassigned physicians' appeal of HCFA reimbursements. PMID- 11066523 TI - Pharmaceuticals: manufacturers' price-fixing schemes challenged. PMID- 11066524 TI - Reproductive health: Eighth Circuit waives certificate of need for planned parenthood. PMID- 11066525 TI - Putting free PSA into play--why and how. PMID- 11066526 TI - Pharmacy face-off over patient testing. PMID- 11066527 TI - Of all analyzers, immunoassay the trickiest. PMID- 11066528 TI - Quality chief vows no 'naming, blaming, shaming'. PMID- 11066529 TI - Nipping contamination in the blood. PMID- 11066530 TI - Net gain: moving labs to the Web. PMID- 11066531 TI - Cytokines' steady march to labs. PMID- 11066532 TI - Thin to thick to thin again? PMID- 11066534 TI - Putting blood in its place. PMID- 11066533 TI - Diagnosing heparin-induced thrombocytopenia. PMID- 11066535 TI - Colon cancer markers still in limbo. PMID- 11066536 TI - Breaking bottlenecks at the front end. PMID- 11066537 TI - Partial drawback: iffy APTTs lead to tube's exit. PMID- 11066539 TI - Quest for connectivity spawns consortium. PMID- 11066538 TI - Bioelectronic chips could be big business for small firm. PMID- 11066540 TI - An array of sizes, prices for the low-volume site. PMID- 11066541 TI - Getting the skinny on SCORES: what you should know. PMID- 11066542 TI - Why PathPAC should not provoke. PMID- 11066543 TI - A first--but pivotal--step for in vitro diagnostics. PMID- 11066544 TI - For HEDIS to succeed, MCOs must get a read on labs' needs. PMID- 11066545 TI - Under the skin: sorting through the hype and hope for noninvasive POC devices. PMID- 11066546 TI - Chemistry analyzers: weigh service, menu, and expandability. PMID- 11066547 TI - From nurses, POC testing gems. PMID- 11066548 TI - Moving blood gas testing to the point of care. PMID- 11066549 TI - In vitro blood gas analyzers. PMID- 11066550 TI - FDA 'not backing away' from universal leukoreduction. PMID- 11066551 TI - Infant abduction in the hospital setting. PMID- 11066552 TI - Rapid HIV testing in labor and delivery settings. PMID- 11066553 TI - Telephone triage and risk management. PMID- 11066554 TI - Consumer confidence. Software can help ease the RFP burden. PMID- 11066555 TI - Western medicine enters the former Soviet Union. PMID- 11066556 TI - Future exam room. Physicianconnection. PMID- 11066557 TI - Can the Web save disease management? PMID- 11066558 TI - The latest word. A glossary of healthcare information technology terms. PMID- 11066559 TI - Prescribing information:no waiting on the Web. PMID- 11066560 TI - Time for an interface-lift? PMID- 11066561 TI - Healthcare takes a cue from Wal-Mart. Interview by Amy S. Feehan. PMID- 11066562 TI - Healthcare report cards. Online rating systems show how organizations and providers measure up. PMID- 11066563 TI - Tightening the accountability chain. PMID- 11066564 TI - Seeing is believing. Visual integration brings new direction to the clinical workstation. PMID- 11066565 TI - The missing link. Digital subscriber lines help focus resources and break backlog. PMID- 11066567 TI - HL7 in the 21st century. Integrating medical information exchange. PMID- 11066566 TI - Building blocks. An integrated IT system lays the foundation for business growth. PMID- 11066568 TI - Malaysia's 21st century goal: IT-based health gains for one and all. PMID- 11066569 TI - A global perspective of health informatics today and tomorrow. Interview by Barbara Hellelgrave. PMID- 11066570 TI - Next generation networks. PMID- 11066571 TI - CIOs speak out. PMID- 11066572 TI - Going interactive. Once the laggards of interactivity, hospitals are beginning to build Web sites consumers can use. PMID- 11066573 TI - Site design tips. The right Web tools mean more business for healthcare organizations willing to invest the time. PMID- 11066574 TI - Spotlight. Managed care information systems. PMID- 11066575 TI - In support of stem cells. PMID- 11066576 TI - The diabetes explosion. PMID- 11066577 TI - Bugging asthma. Minor infections picked up in day care may keep kids from developing the disease. Here's why. PMID- 11066578 TI - Growing pains. What happens when puberty comes too soon in your child--and what you can do about it. PMID- 11066579 TI - High five for a new hand. PMID- 11066580 TI - Too much sun? John McCain's second bout with melanoma reminds all of us to take skin cancer seriously. PMID- 11066581 TI - My day at the clinic. PMID- 11066582 TI - DQIP measures now put providers on same page in gauging quality diabetes care. Diabetes Quality Improvement Project. PMID- 11066583 TI - Encouraging a proactive approach to managing diabetes. PMID- 11066584 TI - An American epidemic--diabetes. PMID- 11066585 TI - Helping to break bad habits. PMID- 11066586 TI - Is your cell really safe? PMID- 11066587 TI - When living is a fate worse than death. PMID- 11066588 TI - The HIV disbeliever. PMID- 11066589 TI - Cloning pigs for parts. PMID- 11066590 TI - Up in smoke: the medicinal marijuana debate. PMID- 11066591 TI - Fragmented bodies, legal privilege, and commodification in science and medicine. PMID- 11066592 TI - Banning partial-birth abortion: drafting a constitutionally acceptable statute. PMID- 11066593 TI - Are you case-managing your hospital out of business? PMID- 11066594 TI - Prevention program cut patient falls by 10%. PMID- 11066595 TI - Practice guideline tackles fall risk. American Medical Directors Association. PMID- 11066597 TI - Clip these tips to reduce falls and eliminate fear. PMID- 11066596 TI - Help patients recover from the fear of falling. PMID- 11066598 TI - Ode to Ruth. A caregiver and resident become girlfriends. PMID- 11066599 TI - Documentation hot spots. Capture the right information and boost reimbursement. PMID- 11066600 TI - Don't overlook today's growth potential. PMID- 11066601 TI - The shift to palliative care. New model can improve quality of life in long-term care. PMID- 11066602 TI - Ka-ching! Are wound care costs clouding clinical judgment? PMID- 11066603 TI - Care continuum. Setting the PACE. Program of All-Inclusive Care for Elderly. PMID- 11066604 TI - Improving drug therapy. Interview by Sam Adler. PMID- 11066605 TI - Hyperlink headaches and other Web woes. PMID- 11066606 TI - Finding funding in 2000. PMID- 11066607 TI - Room service. Another success story in the making. PMID- 11066608 TI - Why carry malpractice insurance? PMID- 11066609 TI - Expect more unannounced surveys. The Joint Commission and HCFA increase the number of surprise visits. PMID- 11066610 TI - Stop and assess your liability status. PMID- 11066611 TI - Financing critical care medicine in 2010. PMID- 11066612 TI - Hospital collections from emergency department admissions. PMID- 11066613 TI - The danger of being too lean. Warning signs that your health care organization has cut too much. PMID- 11066614 TI - Physician assistants as inpatient caregivers. A new role for mid-level practitioners. PMID- 11066615 TI - The consultative skill imperative. Taking a consultative approach to the changing "staff specialist" role. PMID- 11066616 TI - Can you defend your pulse oximeter? PMID- 11066617 TI - Impact of mailing information about nonurgent care on emergency department visits by Medicaid beneficiaries enrolled in managed care. AB - CONTEXT: Emergency department services may be used more appropriately if laypeople's knowledge of managing minor medical problems could be enhanced, especially since Medicaid applies a "prudent layperson" standard for providing access to emergency care. OBJECTIVE: To investigate the effect of mailing a booklet, First Look, that informed Medicaid beneficiaries about care of common nonurgent conditions and encouraged use of alternatives to emergency care including care by office-based physicians, telephonic nursing assistance, and self-care. STUDY DESIGN: A randomized, parallel group study. PATIENTS AND METHODS: Administrative data from 2 health plans serving urban Medicaid populations were used to identify households with a history of emergency department utilization (n = 3101 and n = 3822). Within each health plan, households were randomly assigned to receive First Look. The number of emergency department visits during 6.5 months of follow-up was the primary study endpoint. RESULTS: Compared with controls, 1% fewer members of households that were mailed First Look visited an emergency department in each health plan (23% versus 24% in Plan A; 27% versus 28% in Plan B). The 95% confidence intervals on the observed differences were -3% to 1% and -4% to 1% in Plans A and B, respectively. The proportion of emergency department visits for conditions discussed in First Look was not significantly reduced in households that were mailed the booklet (62% versus 60% in Plan A and 51% versus 48% in Plan B). CONCLUSION: Mailing First Look to Medicaid beneficiaries did not have a significant effect on use of emergency departments. Medicaid programs need to evaluate other, perhaps more multifaceted, interventions to promote appropriate use of emergency departments. PMID- 11066618 TI - "We got mail": electronic communication between physicians and patients. AB - E-mail has the potential to improve both the quality and efficiency of healthcare service delivery. Despite the substantial growth of this form of communication over the past decade, its promise to patients, providers, and their health plans remains largely untapped. In this article we (1) review the literature on e-mail use between patients and providers; (2) identify challenges and opportunities facing managed care organizations that wish to maximize the potential of this form of communication; (3) describe the components of 2 systems aimed at enhancing e-mail use in clinical settings; and (4) discuss the implications of increased e-mail use for managed care. PMID- 11066619 TI - Fairness and rationing implications of medical necessity decisions. AB - When healthcare coverage entails medical necessity review, patients, providers, payers, and government agencies must confront issues of fairness and rationing. To explore the ethical ramifications of medical necessity decisions, we provide 2 illustrative case. In the first case, we discuss the implications of rule-based rationing and in the second we consider the influence of a medical group's internal review council on decisions of medical necessity. Both case examples illustrate why there are no agreed-on rules for setting a threshold for approving or denying care based on medical necessity and suggest that more complex medical cases require a more complex review process. PMID- 11066620 TI - Using administrative data to compare the relative effectiveness of amlodipine vs nifedipine CC. AB - OBJECTIVE: To describe an approach for using claims data to compare the effectiveness of 2 similar drugs used for similar indications within a health maintenance organization. STUDY DESIGN: A database study comparing the effectiveness of amlodipine and nifedipine CC in the initial treatment of hypertension. PATIENTS AND METHODS: The claims records of Pennsylvania Medicaid patients between 18 and 64 years of age with continuous eligibility in 1994 were studied. Pharmacy, hospital, and outpatient claims data were merged, and adult patients receiving the target drugs for the specified indication were identified. The effectiveness of the 2 agents used were compared based on the concept that a change in dispensed medication suggested either an adverse event or lack of effectiveness. Adherence rates, adverse events, and pharmacy and nonpharmacy costs associated with the 2 agents were also compared. RESULTS: Patients receiving amlodipine and nifedipine CC as initial treatment for hypertension had similar demographic characteristics and numbers of comorbid conditions. More patients started on nifedipine CC switched to another calcium channel blocker (15.8% for nifedipine CC vs 10.3% for amlodipine). More patients started on amlodipine switched to another class of antihypertensive agent (13.2% for amlodipine vs 7.3% for nifedipine CC). Patients in both groups received adjunctive antihypertensive drugs at a similar frequency (35% for nifedipine CC vs 42%, for amlodipine). Rates of adherence were similar. In adherent patients, there was no difference in rates of reported adverse events. The nonpharmacy costs were similar between groups. Patients in the amlodipine group also had a trend toward higher overall pharmacy charges (all medications) and higher charges for antihypertensive medications other than the study drugs ($302 vs $188, P = .054). CONCLUSIONS: Claims data are often the best available evidence for comparing the effectiveness of pharmaceuticals in real clinical practice. While these comparisons have inherent limitations, the accuracy of the assessment can be maximized by limiting the assessment to agents with the same specific indications. Other important elements include comparison of crossover rates to other pharmaceuticals in the same class; rates of addition of other pharmaceuticals in the same class, adherence, adverse events, and overall healthcare charges. PMID- 11066621 TI - Use of an open-ended question to supplement a patient satisfaction questionnaire in a medical residents' clinic. AB - OBJECTIVES: To determine (1) the proportion of responses to an open-ended question related to patient satisfaction that could be categorized into 1 or more of 9 previously developed domains of out-patient care and (2) whether any other important aspects of care could be identified by adding the open-ended question to a satisfaction questionnaire. STUDY DESIGN: A 3-month observational study was done at the internal medicine clinic of an urban teaching hospital. PATIENTS AND METHODS: As part of a patient satisfaction study, 511 visitors were asked after their visit, "What are the 1 or 2 things that are most important to you when you see a doctor?" The responses were categorized independently by 2 raters into 1 or more of the 9 domains. When these 2 raters disagreed, the responses were read to a third rater. When either all 3 raters disagreed, or at least 1 rater thought a new domain was mentioned, those responses were categorized by consensus. Interobserver reliability between raters 1 and 2 was calculated by using Cohen's kappa statistic. RESULTS: The 355 responses were categorized as follows: 303 (85.4%) identified one or more domains that were part of the previously developed taxonomy, 9 (2.5%) identified a new domain, 11 (3.1%) identified both old and new domains, and 32 (9.0%) could not be categorized. Cohen's kappa was 0.57 (P < .001). Cultural sensitivity and physician honesty were the additional domains identified, by 1.1% and 4.5% of respondents, respectively. CONCLUSIONS: The previously developed taxonomy of domains can be used in this setting to categorize the large majority of open-ended responses. Such responses can identify important aspects of care that were either previously unidentified or were already identified but given low ratings. This information then can help improve quality of care. PMID- 11066622 TI - Diagnostic testing for influenza: review of current status and implications of newer treatment options. AB - AUDIENCE: This article is intended for all clinicians caring for patients at risk for or infected with influenza. GOAL: To review the accuracy and utility of diagnostic tests for influenza and to compare them with clinical diagnosis. OBJECTIVES: 1. Discuss the epidemiology and scope of influenza infection. 2. Discuss available diagnostic tests for influenza. 3. Discuss the implications of newer options for prevention and therapy. PMID- 11066623 TI - Use of hospital emergency departments for nonurgent care: a persistent problem with no easy solutions. PMID- 11066624 TI - Electronic patient-centered communication: managing risks, managing opportunities, managing care. PMID- 11066625 TI - Staffing models: nurse shortage spurs hunt for perfect ratios. PMID- 11066626 TI - Patient flow QI program garners Codman Award. PMID- 11066627 TI - Benchmarking takes a back seat in home care. PMID- 11066628 TI - ZLUH eases patients' journey through the system. Zale Lipshy University Hospital. PMID- 11066629 TI - Medication errors present benchmarking headache, no consistent measures. PMID- 11066630 TI - Home care experts split on value of OASIS data. PMID- 11066631 TI - Minnesota medical groups share best practices. PMID- 11066632 TI - Program makes assessing competence easier. PMID- 11066633 TI - Perspectives. Gore: more funds for Medicare, incremental steps for access. PMID- 11066634 TI - Perspectives. House Medicare Rx bill: doing what it takes for choice? PMID- 11066635 TI - Marketplace. Hawaii finds business hasn't gotten ready for its comprehensive medical privacy law. PMID- 11066636 TI - Perspectives. Frayed connections: can funding health centers save the safety net? PMID- 11066637 TI - Perspectives. Disparities a shadow on US health landscape. PMID- 11066638 TI - Marketplace. New court ruling in HMO disputes says plans must do what docs want. PMID- 11066639 TI - Perspectives. A long, long trail a-winding into the land of our genes. PMID- 11066640 TI - Supreme Court refuses to "precipitate the upheaval" in Pegram v. Herdrich. PMID- 11066641 TI - The tax-exempt board: fiduciary duties in the context of evaluating strategic alternatives and sales transactions. PMID- 11066642 TI - Analysis and recommendations on the draft OIG compliance program for individual and small group physician practices. PMID- 11066643 TI - Telehealth or telehype? Some observations and thoughts on the current status and future of telehealth. PMID- 11066644 TI - An enhanced healthcare platform via e-medicine. PMID- 11066645 TI - Speech recognition technology: an outlook for human-to-machine interaction. AB - Speech recognition, as an enabling technology in healthcare-systems computing, is a topic that has been discussed for quite some time, but is just now coming to fruition. Traditionally, speech-recognition software has been constrained by hardware, but improved processors and increased memory capacities are starting to remove some of these limitations. With these barriers removed, companies that create software for the healthcare setting have the opportunity to write more successful applications. Among the criticisms of speech-recognition applications are the high rates of error and steep training curves. However, even in the face of such negative perceptions, there remains significant opportunities for speech recognition to allow healthcare providers and, more specifically, physicians, to work more efficiently and ultimately spend more time with their patients and less time completing necessary documentation. This article will identify opportunities for inclusion of speech-recognition technology in the healthcare setting and examine major categories of speech-recognition software--continuous speech recognition, command and control, and text-to-speech. We will discuss the advantages and disadvantages of each area, the limitations of the software today, and how future trends might affect them. PMID- 11066646 TI - Improving the delivery of care and reducing healthcare costs with the digitization of information. AB - In the coming years, the digitization of information and the Internet will be extremely powerful in reducing healthcare costs while assisting providers in the delivery of care. One example of healthcare inefficiency that can be managed through information digitization is the process of prescription writing. Due to the handwritten and verbal communication surrounding prescription writing, as well as the multiple tiers of authorizations, the prescription drug process causes extensive financial waste as well as medical errors, lost time, and even fatal accidents. Electronic prescription management systems are being designed to address these inefficiencies. By utilizing new electronic prescription systems, physicians not only prescribe more accurately, but also improve formulary compliance thereby reducing pharmacy utilization. These systems expand patient care by presenting proactive alternatives at the point of prescription while reducing costs and providing additional benefits for consumers and healthcare providers. PMID- 11066647 TI - Health-risk-assessment tools used to predict costs in defined populations. AB - With the Balanced Budget Act of 1997 mandating that the Health Care Financing Administration (HCFA) implement risk-adjusted payment mechanisms for Medicare managed care plans (Medicare + Choice) by January 2000, risk-adjustment tools will play an important role in future capitated reimbursement. This is because there is growing evidence that healthier-than-average beneficiaries select Medicare + Choice. The risk adjustment that HCFA has adopted is initially based on primary inpatient diagnosis from hospitalizations in the previous year. Other payers are likely to adopt similar payment mechanisms. This article reviews nineteen risk-adjustment research papers, including the tool adopted for Medicare + Choice, some of which are likely to form the basis for subsequent HCFA risk adjustment methods. In general, claims-based models are more powerful in predicting total costs than survey-based or demographics-based models. Survey based models, although expensive and not as powerful claims-based models, can be used when claims data are unavailable. One of the most popular survey-based tools, SF-36, is likely to become increasingly important because HCFA will be using it to measure quality outcomes from Medicare + Choice plans and will make the results public. All of the models reviewed have limitations, but can be expected to be building blocks for future risk-based capitated reimbursement. PMID- 11066648 TI - Leveraging technology for success. AB - Many new information systems, technologies, and inventions promise significant benefit and value. Balancing the ever-increasing technology demands and their related expense with provision of high quality cost-effective patient care is one of today's significant organizational challenges. When appropriately deployed, Information Technology can support organizations in achieving their competitive leverage, market position, quality patient care, and efficient operations. This article presents an overview of the impact of new and emerging technologies, and suggests ways organizations can successfully leverage information systems and technology. PMID- 11066649 TI - Healthcare applications of knowledge discovery in databases. AB - Many healthcare leaders find themselves overwhelmed with data, but lack the information they need to make informed decisions. Knowledge discovery in databases (KDD) can help organizations turn their data into information. KDD is the process of finding complex patterns and relationships in data. The tools and techniques of KDD have achieved impressive results in other industries, and healthcare needs to take advantage of advances in this exciting field. Recent advances in the KDD field have brought it from the realm of research institutions and large corporations to many smaller companies. Software and hardware advances enable small organizations to tap the power of KDD using desktop PCs. KDD has been used extensively for fraud detection and focused marketing. There is a wealth of data available within the healthcare industry that would benefit from the application of KDD tools and techniques. Providers and payers have a vast quantity of data (such as, charges and claims), but not effective way to analyze the data to accurately determine relationships and trends. Organizations that take advantage of KDD techniques will find that they offer valuable assistance in the quest to lower healthcare costs while improving healthcare quality. PMID- 11066650 TI - Universal broadband communications creates new options for healthcare networks. AB - Summary of broad band communications technologies. Definition and explanation of common technologies. Description of how technologies can be applied in multiple different healthcare environments. Analysis of current capabilities, near term improvements and long term trends. Economic analysis of technology choices and their impact on healthcare cost structures. PMID- 11066651 TI - The use of XML in healthcare information management. AB - Extensible Markup Language (XML) is an emerging Internet standard that is gaining momentum in many industries, including healthcare. This article examines the origins of XML, its components, and some potential uses for XML in the healthcare industry. It then discusses a specific initiative to use XML as the basis for an industry-standard scheduling protocol. PMID- 11066652 TI - Electronic commerce: beyond the euphoria. AB - As the center of considerable media attention, case study articles, vendor research, and development efforts, electronic commerce technology is entering healthcare and having a profound effect. The simple truth, however, is that after the drama and excitement begins to wear off, completing a successful e-commerce implementation remains good old-fashioned hard, sometimes monotonous work. To be successful, e-commerce technologies must be planned and implemented with rigorous project standards, and incorporated with significant process and workflow reengineering to actually return significant value to the organization. This article briefs readers on the organizational issues they must consider in evaluating e-commerce readiness--cultural, executive and technological factors that either support or inhibit project and technology success. Readers will be left with the tools to conduct an electronic commerce "readiness assessment" to evaluate the immediate, mid- and long-term potential of electronic commerce; practical remediation strategies for better preparing the organization for the changes inherent in moving to an e-commerce-enabled business model; and comments from the field--advice from organizations that have successfully implemented e commerce technologies into their ongoing operations. PMID- 11066653 TI - Self-benchmarking improves care, cuts drug errors. PMID- 11066654 TI - ACG case-mix system aids contract assessment, improves performance. AB - The ABCs of ACGs. Adjusted clinical groups, or ACGs, a risk adjustment system developed by Baltimore-based Johns Hopkins University, are being used by managed care plans, hospital-based integrated delivery systems, and large provider groups to assess the illness burden of patients for actuarial, payment, profiling, and quality purposes. Find out what ACGs are, how they work, and how to use them. PMID- 11066655 TI - Hospital discharge survey offers LOS, procedure data. AB - Data Library: The National Hospital Discharge Survey offers excellent benchmarking data on inpatient length of stay, demographics, and diagnosis and procedure rates. Review some of the results from the National Center for Health Statistics. PMID- 11066656 TI - Merging lab, drug data boosts disease management. AB - Merging lab and pharmacy data can benefit disease management. Integrating lab data with other data sets gave Blue Cross Blue Shield of Southeast Michigan a way to focus its diabetes disease management program, assess the effectiveness of therapy, and profile physicians. Learn how the plan's experience in data integration can apply to hospital systems and why pursuing a partner in such a project might be a wise move. PMID- 11066657 TI - Hospitals cut CHF readmissions with benchmarking. PMID- 11066658 TI - Readmission analysis leads to heart failure intervention that cuts returns to hospital. PMID- 11066659 TI - Use DRG data to develop reimbursement strategies, improve hospital efficiency. PMID- 11066660 TI - Health system finds sharing data with doctors produces practice changes. PMID- 11066661 TI - Charges, LOS vary widely for cardiac catheterizations amid rising incidence. PMID- 11066662 TI - Patent fight sparks biological warfare in the drug industry. PMID- 11066663 TI - Medicare competitive bidding: an idea stuck in the mud(dle). PMID- 11066664 TI - What are IDSs doing about poorly performing physician networks? AB - A national survey of 105 healthcare organizations conducted by Longshore & Simmons found that most IDSs' networks of acquired physician practices are losing money. The survey examined a variety of issues related to management and structure of IDS physician networks and physician compensation. In addition to finding that most networks were unprofitable, the survey found that for 75 percent of respondents, capitation revenue represented 25 percent or less of total revenue. The survey also disclosed there was no single, preferred method for allocating capitation revenue among physicians; 43 percent of physician networks were organized as hospital departments, cost centers, or divisions; and only 21 percent of respondents were using relative value units as a means to measure physician productivity, despite the proven effectiveness of this approach. PMID- 11066665 TI - Healthcare transition managers make market exits easier for insurers. AB - Insurers have been struggling to meet and maintain their profit margins on the health insurance business they underwrite. As a result, many have exited or are contemplating exiting from the health insurance market. At the same time, managed care organizations are seeking ways to augment their enrollment base and satisfy investor expectations. Insurers that wish to transfer their health insurance business to other carriers and managed care organizations that are interested in accepting this business have turned to a new type of healthcare financing company -the healthcare transition manager--to facilitate the transaction. Part broker, part matchmaker, and part administrative coordinator, these companies make it possible for commercial insurers to exit the health insurance market without undue hardship to enrollees or the managed care plans that assume risk for their care. PMID- 11066666 TI - Lessons learned from three physician-equity models. AB - To improve the profitability of group practice ownership, some healthcare organizations have structured arrangements to include a form of physician equity. An equity incentive is designed to encourage physician behavior that supports business operations by tying financial reward to overall organizational performance. Three physician-equity models--third-party integration, joint venture management services organization (MSO), and physician-owned practice management company--have used the physician equity incentive with varying degrees of success. The experiences of three healthcare systems that implemented these models demonstrates that strategies often cannot be executed as planned, growth should not be assumed, and the changing healthcare marketplace is unpredictable. PMID- 11066667 TI - AICPA standard aids in detecting risk factors for fraud. American Institute of Certified Public Accountants. AB - The American Institute of Certified Public Accountants' Statement on Auditing Standards (SAS) No. 82, Consideration of Fraud in a Financial Statement Audit, requires independent auditors to obtain reasonable assurance that financial statements are free of material mis-statements caused by error or fraud. SAS No. 82 provides guidance for independent auditors to use to help detect and document risk factors related to potential fraud. But while SAS No. 82 suggests how auditors should assess the potential for fraud, it does not expand their detection responsibility. Accordingly, financial managers should discuss thoroughly with auditors the scope and focus of an audit as a means to further their compliance efforts. PMID- 11066668 TI - Using contact capitation to align payment incentives among specialists. AB - Contact capitation is a means of paying specialists based on the number of patients managed rather than on the number of services provided or procedures performed. Payments to physicians are disbursed from budgets, or risk pools, which are established by specialty and product line (e.g., commercial coverage, Medicare). Each specialist is credited with managing a patient for a specified time period (usually 12 months) following the patient's initial visit. To ensure payments are equitable to all physicians, regardless of specialty or subspecialty, the system may be adjusted by using different contact weights for certain diagnoses or procedures, creating subpools for selected subspecialties and/or procedures, establishing separate capitation rates for different age segments, and setting aside certain specialties as fee-for-service carve-outs. Contact capitation has advantages over traditional specialist capitation of removing physicians' financial incentives to overutilize and allowing for a broad physician specialty panel. Challenges to implementing contact capitation include getting physicians to alter habitual practice patterns and managing the system's administrative complexity. PMID- 11066669 TI - Net one, net two: the primary care network income statement. AB - Although hospital-owned primary care practices have been unprofitable for most hospitals, some hospitals are achieving competitive advantage and sustainable practice operations. A key to the success of some has been a net income reporting tool that separates practice operating expenses from the costs of creating and operating a network of practices to help healthcare organization managers, physicians, and staff to identify opportunities to improve the network's financial performance. This "Net One, Net Two" reporting allows operations leadership to be held accountable for Net One expenses and strategic leadership to be held accountable for Net Two expenses. PMID- 11066670 TI - Providers prevail in prudent-buyer therapy litigation. PMID- 11066671 TI - PFS outsourcing partnerships can improve performance and control service costs. PMID- 11066672 TI - Automated receiving speeds deliveries and reduces errors. PMID- 11066673 TI - Life insurance check-ups are necessary. PMID- 11066674 TI - Data trends. Key organizational performance indicators. PMID- 11066675 TI - A new roadmap for professional advancement. PMID- 11066676 TI - Clarifying professional practice. PMID- 11066677 TI - Bracing for the year 2000 Medicare+Choice bug. PMID- 11066678 TI - IDS CFOs need to be flexible: an interview with Phyllis Cowling. AB - In most integrated delivery systems (IDSs) today, the CFO plays a major role in managing a broad range of strategic and operational issues that are integral to the success of the overall organization. To gain insight into the types of challenges that typically confront IDS financial managers, HFM spoke with Phyllis A. Cowling, FHFMA, CPA, vice president and CFO of Baptist St. Anthony's Health System, Amarillo, Texas, and a member of HFMA's Board of Directors. Baptist St. Anthony's (BSA) is a recently formed IDS comprising a full-service hospital with emergency center, four primary care centers, a home health agency, a hospice service, a rehabilitation and skilled nursing facility, a senior health center, an ambulatory surgery facility, and a PPO serving northern Texas, western Oklahoma, and surrounding areas. BSA also has ownership interest in two provider based HMOs. PMID- 11066679 TI - Provider-owned health plans: El Dorado or Armageddon? AB - The development of provider-owned health plans continues to be an important strategy of integrated delivery systems (IDSs). While HMO enrollment growth has continued, reaching almost 70 million people, average health plan profit margins have declined from 8 percent in 1994 to less than 1 percent in 1997. About 56 percent of HMOs lost money in 1998. The ability to successfully develop and operate a provider-owned HMO is affected by conditions inherent to the managed care industry, the level of cooperation among IDS business, units, and local market conditions. PMID- 11066680 TI - Risk pools: payers and providers take the plunge. AB - Providers considering managed care risk pool arrangements should understand thoroughly what services the pool covers, the time period covered, and how the pool is administered. Important issues related to pool administration include how credits and debits are applied; when the accounting occurs (interim, year-end, or contract termination); and provisions for reports, audit rights, and dispute resolution. Although a pool arrangement gives the health plan control over claims payment, the risk allocation made possible through a pool arrangement helps ensure that the economic incentives of the health plan and the provider are aligned. PMID- 11066681 TI - Realizing the financial benefits of capitation arbitrage. AB - By anticipating the arbitrage potential of cash flow under budgeted capitation, healthcare organizations can make the best use of cash flow as a revenue generating resource. Factors that determine the magnitude of the benefits for providers and insurers include settlement interval, withhold amount, which party controls the withhold, and incurred-but-not-reported expenses. In choosing how to structure these factors in their contract negotiations, providers and insurers should carefully assess whether capitation surpluses or deficits can be expected from the provider. In both instances, the recipient and magnitude of capitation arbitrage benefits are dictated largely by the performance of the provider. PMID- 11066682 TI - Physician unionization efforts gain momentum, support. AB - Physicians increasingly are assuming the status of employees in healthcare organizations. Physicians also are seeing restrictions imposed on their practices by healthcare organizations seeking to control costs of care delivery. These trends have led a growing number of physicians to attempt to organize into unions. Obstacles to physician unionization efforts have included Federal antitrust laws that prohibit physicians from organizing, as well as physician reluctance to engage in organized activities they see as antithetical to their professional duties (e.g., strikes). In addition, physicians' attempts to unionize frequently have failed due to provisions of the National Labor Relations Act, which authorize collective bargaining only among individuals designated as "employees." Physicians seeking to form unions often are thwarted by the argument that they are not employees, but rather students, independent contractors, or supervisors, and therefore not entitled to protection under the act. Nonetheless, a number of recent developments, such as the American Medical Association's decision to endorse unionization by physicians and the National Labor Relations Board's decision that attending physicians should be regarded as employees, not supervisors, are creating a climate more conducive to physician unionization in the United States. PMID- 11066683 TI - Using an effective business model for group practice management. AB - Managing a group practice effectively can improve the practice's bottom line as well as attract a capital partner, if necessary. By addressing issues such as culture, values, governance, role definition, and expectations, group practices can clarify their vision and goals and run their business in an organized, efficient manner. When a group practice's physicians are committed to the success of the practice, they can work as a team to implement efficient operational procedures and optimize revenues. Effective business model components should be considered by both fledgling and mature practices. PMID- 11066684 TI - Understanding Medicare+Choice risk adjustment. PMID- 11066685 TI - What to expect when negotiating a CFO compensation package. PMID- 11066687 TI - Planning a value-added Web site. PMID- 11066686 TI - Choosing an investment management approach for bond issue proceeds. PMID- 11066688 TI - State college savings plans offer tax breaks. PMID- 11066689 TI - Data trends. Hospitals experiencing new wave of wage pressures. PMID- 11066690 TI - Enhancing what you know through who you know. PMID- 11066691 TI - Getting billing right. PMID- 11066692 TI - Campaign 2000: insuring the uninsured. PMID- 11066693 TI - Annual audits of IDS risk contract settlements improve payment accuracy. AB - Integrated delivery systems (IDSs) should conduct annual audits of payers' settlements under risk contracts to verify that the payer attributed the appropriate amounts of revenue and charged the appropriate claims expenses to the IDS. In particular, IDSs should verify that payers calculated revenues and expenses based on consistent member counts and that the determined commercial revenue was based on the actual premiums paid. IDSs also should determine whether payers have used appropriate demographic factors and countywide rates as a basis for determining Medicare revenue, charged the IDS for claims only for valid members, paid capitated providers the correct capitation amounts, and used appropriate historical data to estimate the amounts of incurred-but-not-reported claims attributed to the IDS. PMID- 11066694 TI - Four methods of setting group premium rates require different insurer resources. AB - Methods used to calculate premium rates for group health insurance coverage require varying levels of staff and financial commitment and differ in how closely they match premium revenue to projected expenses in aggregate. Determining the appropriate method for a group requires an analysis of resources available as well as an examination of competitive forces, state and Federal legislation, and underwriting issues. PMID- 11066695 TI - Providing capital for physician group practices: new opportunities for hospitals. AB - As physician group practices grow and consolidate, they have an increasing interest in developing close capital partnerships to ensure access to capital. Yet as many healthcare organizations have sought to divest poorly performing acquired physician practices, physicians have seen their pool of potential capital partners shrink. Under these conditions, hospitals have a new opportunity to present themselves to physician group practices as attractive capital partners. To understand the nature of this opportunity, one needs to know why group practices seek capital, how groups approach their investment strategies, and what criteria they use to compare prospective capital partners. To build stronger relationships with physicians, hospitals should focus on turning around their poorly performing acquired physician practices and pursue strategies such as collaborating with physician practice management companies and developing new models for partnering with physicians (e.g., special purchase agreements and more advanced management services organizations). PMID- 11066696 TI - IRS issues new disclosure rules for tax-exempt organizations. AB - The IRS earlier this year issued new regulations that tax-exempt organizations must follow in disclosing to the public certain documents as directed by the Omnibus Budget Reconciliation Act of 1987 (OBRA87) and the Taxpayer Bill of Rights 2 of 1996. The new rules went into effect June 8, 1999. Complying with these rules may pose difficulties in terms of the increased exposure of the documents that must be disclosed, increased labor, time limitations, and penalties, some of which can be imposed upon a director or officer. An organization can escape some of the burden of responding to requests by posting the documents on its Web site, but it still must make the documents available for inspection at its principal office and certain regional offices. PMID- 11066697 TI - Debt-maturity structures should match risk preferences. AB - Key to any debt-maturity matching strategy is financing assets with the appropriate debt structure. Financial managers need to establish an optimal capital structure and then choose the best maturity-matching structure for their debt. Two maturity-matching strategies that are available to healthcare financial managers are the accounting approach and the finance approach. The accounting approach, which defines asset maturities as current or fixed, is a riskier financing strategy than the finance approach, which defines asset maturities as permanent or temporary. The added risk occurs because of the accounting approach's heavy reliance on short-term debt. The accounting approach offers the potential for lower costs at the expense of higher risk. Healthcare financial managers who believe the financing function should support the organization's operations without adding undue risk should use the finance approach to maturity matching. Asset maturities in those organizations then should be considered permanent or temporary rather than current or fixed, and the debt-maturity structure should reflect this. PMID- 11066698 TI - Careful telemedicine planning limits costly liability exposure. AB - Recent Federal and state legislation and new payment opportunities from Medicare, Medicaid, and private payers may make it possible to offer telemedicine as a viable, cost-effective alternative to traditional care delivery in communities where access to health care is limited. Originally, nonexistent payment and expensive technology held back telemedicine but, these barriers are giving way to specific applications that can yield dramatic cost savings for group practices in the delivery of medical care while adding features and benefits not typically available in traditional delivery settings. Before joining a telemedicine network, group practices need to negotiate a variety of legal issues related to the corporate practice of medicine, patient confidentiality and privacy, malpractice, informed consent, licensure and credentialing, intellectual property, Medicare and Medicaid payment, fraud and abuse, medical device regulation, and antitrust. PMID- 11066699 TI - Confusion in health laws confound providers' quest to comply. PMID- 11066700 TI - Diversification results in steady returns. PMID- 11066701 TI - Data trends. Burden-of-illness comparisons: a tool for managing risk contracts. PMID- 11066702 TI - BBA bad news gets worse, thanks to flawed calculations. PMID- 11066703 TI - How IDSs can turn BBA postacute care provisions to their advantage. AB - Reductions in payments imposed by the Balanced Budget Act of 1997 may force postacute care providers to impose limits on the number and acuity of patients they can accept. As a result, integrated delivery systems may face reduced access to postacute care. An integrated delivery system's financial well-being may be undermined if its only alternative is to care for postacute care patients in high cost, acute care settings. To address this problem, IDSs should analyze the financial impact of the Balanced Budget Act and share results throughout their systems, determine how financial incentives affect postacute care utilization, conduct interviews to garner support for strategic objectives, evaluate current operational policies and procedures to determine whether they meet Balanced Budget Act requirements, assess their demand for postacute care services, and develop strategies that fairly distribute the impact of changes among all constituencies. PMID- 11066704 TI - Optimizing revenues through effective contract management. AB - Many provider organizations are losing millions of revenue dollars each year because their systems and administrative processes do not efficiently and effectively monitor and enforce contract terms. This inefficiency may result from many factors, including lack of understanding of managed care contracts, terms, and requirements; limited information systems; and inadequate operational controls that interfere with an organization's ability to capitalize on contract terms. Adequate information systems, process improvements, and well-informed contract management staff can help many organizations reduce costly errors and inefficiencies that result when the negotiated terms of managed care contracts are not enforced. PMID- 11066705 TI - Minimizing antitrust exposure in a virtual merger. AB - As an alternative to complete mergers or joint ventures, hospitals recently have begun to explore virtual mergers, in which the parties are able to retain some managerial and financial independence while coordinating their mutual operations to financial advantage. Because virtual mergers are a recent phenomenon and can be structured in various ways, the antitrust risks associated with such transactions are unclear. A state antitrust challenge brought against an East Coast virtual merger and informal guidance by Federal antitrust attorneys suggest that the antitrust agencies will be inclined to challenge a virtual merger if the parties to the transaction retain too much independent decision-making authority. Hospitals that are considering a virtual merger therefore would do well to structure the transaction to combine governance and administration, financial assets, operations, and medical staffs as much as possible, while still allowing each party to the transaction to retain the independent decision-making authority each feels is necessary. PMID- 11066706 TI - Using activity-based costing to track resource use in group practices. AB - Research shows that understanding how resources are consumed can help group practices control costs. An American Academy of Orthopaedic Surgeons study used an activity-based costing (ABC) system to measure how resources are consumed in providing medical services. Teams of accounting professors observed 18 diverse orthopedic surgery practices. The researchers identified 17 resource-consuming business processes performed by nonphysician office staff. They measured resource consumption by assigning costs to each process according to how much time is spent on related work activities. When group practices understand how their resources are being consumed, they can reduce costs and optimize revenues by making adjustments in how administrative and clinical staff work. PMID- 11066707 TI - For-profit chains seek to acquire successful not-for-profit hospitals. AB - Over the last decade, investor-owned hospital corporations have grown primarily by acquiring other for-profit hospital chains or stand-alone for-profit institutions. Between 1990 and 1995, however, these corporations also acquired nearly 50 not-for-profit hospitals and converted them to for-profit status. An examination of the long-term financial condition of 39 not-for-profit hospitals acquired by various investor-owned hospital corporations between 1992 and 1996 was conducted using free-cash-flow accounting valuation. The results suggest that, initially, only not-for-profit hospitals in dire financial straits were candidates for acquisition and conversion to for-profit status. More recent acquisitions increasingly have involved more successful not-for-profit hospitals. PMID- 11066708 TI - Issues in group practice management: an interview with Craig Faerber. PMID- 11066709 TI - Federal government increases oversight of nursing homes. PMID- 11066710 TI - Improving job interview skills. PMID- 11066711 TI - EDI cuts cost of managed care contract administration. PMID- 11066712 TI - Data trends. Key organizational performance indicators. PMID- 11066713 TI - How consumers evaluate health care quality: Part I. AB - Information is a critical factor in marketing health care services. Consumers evaluate information according to its source. This report is the first of a three part series examining the way in which Americans evaluate the quality of consumer information as they make decisions on health care products and services. This report provides an overview of the study. The next two installations will focus, respectively, on health plans, hospital and doctors. PMID- 11066714 TI - Message content of alcohol moderation TV commercials: impact of corporate versus nonprofit sponsorship. AB - This content analysis examines a sample of 203 alcohol-related North American TV commercials dealing with alcohol moderation and driving under the influence (DUI), in order to determine whether the type of ad sponsor has an impact on the message content. Corporate sponsors, such as breweries and distillers, are compared to nonprofit sponsors such as governments and nonprofit organizations. Findings show that ads from corporate sponsors are less likely to make mention of threats or negative consequences, and are also less likely to use fear arousal. However, DUI/alcohol moderation ads from corporate sponsors and nonprofit sponsors do not differ in the degree to which they use humor or positive approaches. PMID- 11066715 TI - Key steps in the strategic analysis of a dental practice. AB - As dentistry is becoming increasingly competitive, dentists must focus more on strategic analysis. This paper lays out seven initial steps that are the foundation of strategic analysis. It introduces and describes the use of service customer matrices and location-proximity maps as tools in competitive positioning. The paper also contains a brief overview of the role of differentiation and cost-control in determining key success factors for dental practices. PMID- 11066716 TI - Retail health marketing: evaluating consumers' choice for healthier foods. AB - This study investigates the effect of socioeconomic and demographic variables, nutrition and health related factors, attitudes, and use of nutritional labels on consumers' choice for healthier food products. Seven equations are estimated representing different food types: luncheon meat, milk, cheese, ice cream, salad dressing, dessert, and meats. The results generally indicate that individuals who are less likely to choose a healthier alternative of a food product include: blacks, younger individuals, males, those with smaller households, smokers, those who take less exercise, those who are not on a special diet, those who are less aware about the linkage between diet and disease, those who put more importance on taste when food shopping, and those who less frequently use nutrition panels and labels that describe health benefits on food packages. PMID- 11066717 TI - Perceptions of young adults as to the future of health care in the 21st century. AB - This study assessed the perceptions of the need for and quality of future health care programs of those who will be major users of medical services in the 21st century. Findings indicated that most recognize the importance of having medical coverage. While copayment costs were most important in health plan selection, less personal factors (e.g., ease of obtaining appointments) were more important than the plan's quality reputations and its physicians. Finally, few respondents thought the quality of physicians and hospitals would improve. Even fewer believed the range of services covered would be better, access to care would be easier to obtain, or that service would be more personalized. They also expect the costs of health care to escalate, and costs to have more impact on the availability of medical services. PMID- 11066718 TI - Children's immunizations: the gap between parents and providers. AB - Children's immunizations have been a part of our health care practice for many years. While immunizations have reached record highs in recent years, there is a fear that complacency may cause a drop in immunization rates and thus increases in disease rates. This study explored immunizations from two perspectives. First, focus groups with parents examined immunization information acquisition, practices, and barriers. Next, a physician survey examined immunization provider perceptions of parents' information acquisition, barriers and immunization practices. Several gaps were discovered between these two groups controlling the immunization of our children. Suggestions are made as to possible paths to begin addressing these gaps in order to increase immunization rates. PMID- 11066719 TI - How consumers evaluate health care quality: Part II. AB - This article is the second in a series which examines the way in which consumers assess information regarding the quality of health care services. In the previous article it was demonstrated that, in the view of health care consumers, three major perceptions held by health care consumers, are: (1) substantial differences in quality exist among health care providers, (2) little information is available that allows for the comparison of health care providers on issues related to quality, and (3) when such information is available it is found to be useful and often serves as the basis for decision regarding the choice of health care providers. We further discussed the short coming of marketing strategies based on complex quality indicators and the difficulties of image advertising in an age of institutional mistrust. The reader is reminded that these findings relate to the subjective assessments of consumers, not to objective facts concerning health care delivery. PMID- 11066720 TI - Value of services provided by pharmaceutical companies: perceptions of physicians and pharmaceutical sales representatives. AB - Pharmaceutical sales representatives (PSRs) are a key component of pharmaceutical companies' marketing strategies in that they are the link between the pharmaceutical company and the physician. PSRs provide various services in order to increase the physician's prescribing activity of their companies' products. Given the high cost of recruiting, training, and supporting a PSR, it is important for PSRs to understand the relative significance physicians ascribe to services provided. This study examined whether there is a gap in the perceptions of physicians and PSRs regarding the value of specific services provided by PSRs. Physicians and PSRs who attended medical meetings were surveyed. Results of the study indicated that there were significant differences in the perceived value between PSRs and physicians. Services which were perceived to be less important to physicians than to PSRs were new product detailing, old product detailing, providing product studies and research findings, PSRs serving as expert consultants, and recruiting physicians to participate in FDA approval drug studies. Services for which there were no significant differences of perceived value between the groups included free product samples and promotional luncheons and dinners. PMID- 11066721 TI - Are retail prices "just" when they do not include social costs? AB - The price is "right" when the buyer agrees to purchase goods or service. But is it "just"? That is, does the price include social costs such as pollution and discrimination? Cost shifting is the passing down of these costs from the seller to the buyer. The amount of cost shifting depends upon the inelasticity or elasticity of supply or demands, methods of assigning social costs, means used in promotion and the role of the market price. Wal-Mart and Body Shop International exemplify the problems of social costs. Recommendations for marketing managers concludes the article. PMID- 11066722 TI - A national study: the use of specialty surgical teams. AB - Perioperative nursing is changing in response to the increasing complexity of patient care during diverse, specialized surgical procedures. As a result, designated specialty surgical teams have developed to fulfill needs of patients, nurses and surgeons. This exploratory, descriptive study examined reasons for implementing specialty surgical teams, the frequency and RN composition of such teams and their benefits. Data concerning the prevalence and specific use of specialty surgical teams are necessary to validate and redefine the nature and role of specialized perioperative nurses. Results of the U. S. study generally paralleled the Missouri study although findings were not as pronounced. Clearly though, in both studies, specialty surgical teams are perceived very positively by study respondents. PMID- 11066723 TI - Overcoming barriers to effectiveness in a health care operational environment: building on the lessons of American industry. AB - Several of the manufacturing-based philosophies, techniques and tools, such as Total Quality Management (TQM), Continuous Improvement (CI), Business Process Reengineering (BPR) and Time-based Competition (TBC) have been successfully adapted for use within the service sector. Diverse service industries including airlines, insurance, food services and hospitality have increased customer satisfaction and performance through the use of the quality driven, manufacturing based philosophies. This article explores the reasons for the limited success of TQM/CI, BPR, TBC and benchmarking within the health care industry. Sixteen barriers to change are identified, possible counter-measures to these barriers are outlined and two conceptual frameworks are offered as possible facilitators of change for the health care industry. PMID- 11066724 TI - Musculoskeletal disorders and the consumption of disability products. AB - Although consumers' need assessment generally suggest that, what consumers need as products or services influence their consumption, little research on consumption of disability resources, relating to musculoskeletal disorders exists. However, most medical practitioners defined musculoskeletal disorders by symptoms or clinical disorders and other used self-report. The authors examined the effects of musculoskeletal disorders and ultimate consumption of specific products (orthopedic devices, medications), as well as the cost of surgical options (e.g., gastric reduction surgery). Emphasis was on product categories, such as disability devices and consumption of disability resources. The authors conclude that there are positive relationships between musculoskeletal disorders and consumption of disability products, either to maintain certain lifestyles or reduce pains and sufferings. Finally the authors discussed the advancement of marketing theory with musculoskeletal consumption analysis. Opportunity for further research in the area is enhanced and encouraged. PMID- 11066725 TI - Assessment of an outreach program that links children who use New York City immunization clinics to primary care. AB - BACKGROUND: Many children who obtain immunizations at the New York City (NYC) Department of Health (DOH) Immunization Clinics do not have medical coverage and therefore do not have access to primary care. A consequence of this is increased morbidity of diseases which are best managed in an outpatient setting. In 1996, Every Child By Year 2000 (ECY2000), an outreach program, was launched to educate and link families to primary care and ensure that children are referred to their primary care providers for their health care needs. METHODOLOGY: Families using the NYCDOH Immunization Clinics were interviewed and enrolled in either Child Health Plus (CHP)--a New York State sponsored health insurance program or Medicaid (MA). A survey was designed to determine the number of children, socioeconomic factors of the families that were linked to CHP or MA and factors affecting children's accessibility to primary care and immunization. RESULTS: A total of 4024 children had come into contact with the program in the first half of 1997. A total of 693 children were surveyed and 365 (52.7%) of them were enrolled in CHP or MA. Families who were aware that CHP was a free/low cost health insurance program for children were more likely to have CHP insurance. Children who were enrolled in CHP were more likely to have regular primary care, to receive up-to-date immunization shots and immunization information from their primary providers. CONCLUSION: More public health primary care outreach and education programs should be targeted to recent immigrants and low income families and these programs should be culturally sensitive. PMID- 11066726 TI - Demographic and psychosocial characteristics of cognitively-intact chronically Ill elders receiving home health services. AB - The intent of this study is to sensitize the home health care milieu to the psychosocial needs of home health patients in the effort to initiate effective discharge planning as soon as possible. All of the disciplines that comprise the treatment team play a vital role in relapse prevention; however, the role of the social worker, providing social and emotional interventions to speed recovery and/or prevent exacerbation, has become more prominent. Key demographic and multivariate analysis findings are discussed related to the prevention of relapse. Social work interventions are discussed for several target issues. PMID- 11066727 TI - Predictors of home care readiness for managed care: a multivariate analysis. AB - As home care agencies change from cost-based reimbursement to the managed care risk paradigm, many lack experience with organizational resources and functions needed to successfully operate in the managed care environment. This survey assessed the level of "readiness" for managed care reported by 162 randomly selected home care agencies in three mid-western states. Managed care readiness was measured by a 32-item mailed questionnaire addressing clinical, financial, operational, and informational systems within each agency. Higher levels of readiness were significantly related to the perception of being ready, proprietary auspices, and moderate agency size in a multivariate regression model. Agencies with these characteristics appear to be well positioned as home care moves into the managed care environment. PMID- 11066728 TI - Description and outcomes of a Medicare case management program by nurses. AB - During the current environment of cost-cutting and restructuring, case management is viewed as a means of providing comprehensive, coordinated care while reducing costs. Studies of community-based case management have been disappointing. This study evaluates a Medicare service, Management and Evaluation of the Care Plan (MAE), as a model of nurses providing case management in the home. Three groups were compared: MAE (N = 176), non-MAE (N = 187) and newly discharged (N = 93). Utilization data was collected over one year. Overall, subjects were unmarried older women with three functional limitations, four medical diagnoses and averaged 43 home care visits and one hospitalization. MAE patients were older, had more functional and environmental limitations, less ADL independence, and a worse prognosis, yet used significantly less health care than non-MAE recipients did. Regression analysis was performed using group membership and hospital and home care utilization as dependent variables. Although the project was conducted at one site, overall the sample was similar to the national Medicare population. PMID- 11066729 TI - Moving from Illness to wellness: an important goal of home care. AB - For many clients in home care with chronic illness the real work of getting well involves learning to adjust to chronic illness and to disability. Learning to adjust to disability by maximizing the abilities and qualities that are present can mean overcoming otherwise very significant problems while still maintaining a high quality of life. Home care providers play an important role in helping clients and their families optimize functioning to do the real work of getting "well." PMID- 11066730 TI - Therapeutic recreation treats depression in the elderly. AB - Therapeutic recreation can be an effective method to treat depression in elderly home care patients. Home care is the fastest growing component in the Medicare budget. The co-occurrence of physically limiting conditions and depression in the elderly is well documented. Untreated depression carries an enormous risk and cost. Therapeutic recreation is an ideal psychosocial treatment for use in the home care setting because of its effectiveness and versatility. Certified therapeutic recreation specialists use various interventions such as poetry, music, and exercise as part of a treatment team. In addition to effectively managing depression, therapeutic recreation can be beneficial in reducing the effects of many concurrent physical conditions. PMID- 11066731 TI - Price survey. I.V. prices creep upward again. PMID- 11066732 TI - Strong first year prompts growth push for Consorta. PMID- 11066733 TI - Communication key to supply chain efficiency. PMID- 11066734 TI - 20 challenges of geriatric care. AB - Responding to incidents involving geriatric patients presents multiple challenges and considerations for care. A variety of changes occur to the human anatomy, the physiological functions of the body and emotional state as an individual ages. Understanding these changes, their effect on medication administration and treatment options is important for the EMS provider. The elderly represent a rapidly growing segment of our prehospital patient population. More people will reach age 65 by 2020 than at any other time in the world's history. Many of these individuals will present to the EMS system in need of emergency care and may have multiple chronic conditions that will complicate your patient assessment and available treatment options. Knowing the relationship between aging and a patient's overall health provides the best basis for treating this precious population. PMID- 11066735 TI - Take aim--hit your IO target. A comprehensive approach to pediatric intraosseous infusion, including site selection, needle insertion & ongoing assessment. Part 2. PMID- 11066736 TI - Joint ventures. Prehospital evaluation & treatment of arthritis. PMID- 11066737 TI - Meeting standards challenges in home care settings. PMID- 11066738 TI - Treatment authorization and appeal standards for networks revised. PMID- 11066739 TI - Coordinating sentinel event monitoring with state agencies. PMID- 11066740 TI - Modified sentinel event standard for all manuals. PMID- 11066741 TI - Restraint and seclusion: progress update. PMID- 11066742 TI - Acting in good faith. PMID- 11066743 TI - ORYX data come to life in pre-survey reports. PMID- 11066745 TI - Interim solution permits "weighting" of adverse decisions in tailored surveys. PMID- 11066744 TI - Revised accreditation decision categories and definitions announced. PMID- 11066746 TI - O'Leary testifies before Congressional hearings on patient safety. PMID- 11066747 TI - Public Information Policy revisions increase scope of disclosable information. PMID- 11066748 TI - Plan to implement core measures approved. PMID- 11066749 TI - An examination of how adult and theological education literature informs the practice of clinical pastoral care: a critical review. AB - Examines the literature on adult learning and theological education in relationship to the field of clinical pastoral education. Elicits insight from the source material that informs practice. Evaluates strengths and weaknesses in the literature, particularly in reference to self-directed and transformative learning. Gestures toward implications for pastoral care. PMID- 11066750 TI - The public debate on life and death choices: a response from a Jewish hospital chaplain. AB - Addresses the issue of life and death choices from the perspectives of a Jewish chaplain. Explores the tension between the law (Halachah) and experience or story (Aggadah) and the impact of that tension on lives and decisions. Cites persons and situations from biblical material, oral tradition, and modern times which illustrate the human struggle with these disputed issues. Explores different levels of pain and our understanding of and response to suffering. Invites reflection on the range of responses to pain that are humanly possible. Illustrates how meaning in life nurtures the desire to live. Claims this meaning aspect is not adequately addressed in the current dialogue. Focusses on the role of chaplains in listening and responding to persons and in helping to activate the impulse of hope. Suggests ways in which chaplains can be advocates for patients, prophetic voices within communities and institutions, and empowering of the gifts of others. PMID- 11066751 TI - Levinson's theory of the life structure: the case for its universality. AB - Considers arguments against the universalistic claims of developmental theory, focusing on Daniel Levinson's theory. Argues that a case can be made for the universality of his idea of the life structure if the biological foundations of this idea are considered. Uses Freud's theory of the life and death instincts to support this argument. Concludes that academicians in pastoral care should give as much weight to the individual/biological as the individual/cultural axis of human identity. PMID- 11066752 TI - Judaism, professional intervention, and ethics in long-term care. AB - Discusses spiritual, professional, and ethical approaches to work with an elderly Jewish patient in a nursing home. Addresses the knotty issues of determining decision-making capacity, the refusal of nutrition and hydration, Judaic values pertaining to ending one's life, the role of the spiritual caregiver, the role of the social worker, and the ethical principles of autonomy, beneficence, paternalism, and do no harm. PMID- 11066753 TI - Objects and idols: the significance of internal object relationships for the religious quest. PMID- 11066754 TI - Breaking the power of Ondine's Curse. PMID- 11066755 TI - The impact of lay pastoral telecare on the spiritual well-being of church attenders. AB - Examines the efficacy of Lay Pastoral Telecare (LPT) on the spiritual well-being and church satisfaction of churchgoers (N = 207). Compares an experimental and a control group and concludes that the use of telephone by lay pastoral caregivers can be a means of promoting interpersonal support and enhancement of spiritual well-being within a church congregations. PMID- 11066756 TI - Where there is hope, there is life: toward a biology of hope. AB - Argues the thesis that where there is hope there is life. Grounds this thesis philosophically and theologically, then reviews it from the medical and nursing literature, and illustrates it in a vignette of hospital ministry. Argues that hope can enhance the quality and even the quantity of life. Proposes that hope helps people to deal with their feelings and to cope with their illness. Hope affects immunity and survival. Challenges chaplains, pastoral counselors, and parish clergy to become more effective "agents of hope." PMID- 11066757 TI - A Palm Sunday crucifixion. PMID- 11066758 TI - Pieta: a portrait of forgiveness. PMID- 11066759 TI - The same routine everyday. PMID- 11066760 TI - A happy or a blessed Easter? PMID- 11066761 TI - Hands on ministry: a few lessons from a day as a CNA. PMID- 11066762 TI - We can't let them devalue primary care. PMID- 11066763 TI - After the hurricane, the healing begins. PMID- 11066764 TI - Creating "paths to excellence" in his community. PMID- 11066765 TI - When the doctor's legal history is irrelevant. PMID- 11066766 TI - What's in the box? Is United Healthcare delivering on its promises? PMID- 11066768 TI - A big new break for pension plans. But act fast! PMID- 11066767 TI - What the Texas-Aetna agreement really means. PMID- 11066769 TI - The good news--and bad--about Web-based EMRs. PMID- 11066770 TI - Charges fly in Miami. PMID- 11066771 TI - The triad shuffle. PMID- 11066772 TI - Pa. high court upholds tax exemption. PMID- 11066773 TI - Gaming the system. PMID- 11066774 TI - Troubles for JOAs. PMID- 11066775 TI - Aetna settles with Texas. PMID- 11066776 TI - While the getting's good. PMID- 11066777 TI - HealthAllies.com's new tack. PMID- 11066778 TI - When the checks aren't balanced. W.Va. hospital chief takes a fall. PMID- 11066779 TI - Second opinions on coverage. AB - It's the main complaint about managed care: Doctor and patient agree on a specific treatment, but the HMO refuses to pay for it. Until recently, the only remedy available to patients was long and costly lawsuits. Now, patients and health plans are finding that external review boards--panels of independent experts--offer an inexpensive and fair alternative for resolving such disputes. PMID- 11066780 TI - Some PPMCs succeed where others failed. PMID- 11066781 TI - More relief still a question mark. PMID- 11066782 TI - A venture-capital magnet. PMID- 11066783 TI - Insuring India. Western-style plans will bring big change. PMID- 11066784 TI - Sidestepping the altar. AB - Following several ugly corporate divorces, more hospitals and healthcare systems are deciding to drop their merger plans. They're realizing that it just doesn't make sense to make the trip to the altar, even after long courtships. "Mergers in and of themselves have not been a panacea," one analyst said. "In fact, the reality of mergers has pointed out a lot of shortcomings in the executions, or perhaps in the expectations." PMID- 11066785 TI - Wanted: CEO with entrepreneurial spirit. PMID- 11066786 TI - Alliance launches Blue Cross in Mexico. PMID- 11066787 TI - Making blood safe the world over. PMID- 11066788 TI - Fla. Argentine centers collaborate. PMID- 11066789 TI - Global policy developed in Texas. PMID- 11066790 TI - Seniors in Japan access new services. PMID- 11066791 TI - The universal solution. PMID- 11066792 TI - Venturing into Latin America. PMID- 11066793 TI - New American bites bankruptcy bullet. PMID- 11066794 TI - Petty cash? PMID- 11066795 TI - SEC cracks down on AHERF. PMID- 11066796 TI - The old guard stands down. Generation of CEOs that fostered systems is retiring. AB - It's a changing of the guard. In the past 18 months, some of the nation's best known and longest-serving healthcare chief executives have relinquished the day to-day management of their organizations. And more CEOs have announced their intention to leave this year or next. This is a generation that came of age with the Medicare program and has witnessed the rise of large systems and for-profit healthcare. PMID- 11066797 TI - Not by the numbers, please. PMID- 11066798 TI - Investing solutions. PMID- 11066799 TI - Healthcare IPOs make a comeback. PMID- 11066800 TI - The worst is over? PMID- 11066801 TI - Getting meaningful results from QI monitoring efforts. PMID- 11066802 TI - Nurses uneasy about i.v. sedation for long cases. PMID- 11066803 TI - Deaths after liposuction studied. PMID- 11066804 TI - 'Close the loop' in QI for ASCs. PMID- 11066805 TI - Most are using shielded trocars. PMID- 11066806 TI - ASA updates waste gases advice. PMID- 11066807 TI - What's best for laryngoscope blades? PMID- 11066808 TI - Efficient scheduling of OR cases. PMID- 11066809 TI - Wait for APCs may be ending. PMID- 11066810 TI - Giving O2 halves infection rate. PMID- 11066811 TI - DoD implements changes to third party collection program. PMID- 11066812 TI - Meeting the EMTALA challenge. PMID- 11066813 TI - Building bridges to healthy skin. Nutritional interventions support healing. PMID- 11066814 TI - Building bridges to healthy skin. Stepping up liability prevention practices. PMID- 11066815 TI - Building bridges to healthy skin. Mapping out prevention strategies. PMID- 11066816 TI - Building bridges to healthy skin. Treatment teams maximize healing. PMID- 11066817 TI - Building bridges to healthy skin. Optimizing mobility promotes healthy skin. PMID- 11066818 TI - Providers trapped in survey standoff. PMID- 11066819 TI - Designing for late-stage dementia care. PMID- 11066820 TI - Minimizing cross-contamination. PMID- 11066821 TI - Adverse actions reported nationally. PMID- 11066822 TI - Providers take a PPS reality check. PMID- 11066823 TI - Medication reviews critical in ICFs/MR. PMID- 11066824 TI - Private 'Internets' can unite company. PMID- 11066825 TI - Double your data. A gathering of twins provides a unique research opportunity. PMID- 11066826 TI - Preemie problems. The sobering statistics. PMID- 11066827 TI - Doc shopping? A closer look yields a healthier choice. PMID- 11066828 TI - The perils and promise of Xena. Can cloned pigs yield human transplants? PMID- 11066829 TI - Inefficient, corrupt, and unreliable. The system for testing treatments is in chaos. PMID- 11066830 TI - Wounding with words. Most parents inappropriately lash out at the kids. PMID- 11066831 TI - Could your phone cause cancer? Don't get hung up on it. PMID- 11066832 TI - Baby may have a flat head, but parents shouldn't get too bent out of shape. PMID- 11066833 TI - Miracle cells? Maybe. More research dollars, more ethical quandaries. PMID- 11066834 TI - C-sections rise but may not be the kindest cut. PMID- 11066835 TI - Doctors face a candid camera. PMID- 11066836 TI - Analyzing fire risks building by building. Performance-based fire protection lets architects focus on a building's design objectives, instead of searching for ways to meet generic code provisions. PMID- 11066837 TI - Health-related relief in the former Yugoslavia: needs, demands, and supplies. AB - INTRODUCTION: Many organizations rally to areas to provide assistance to a population during a disaster. Little is known about the ability of the materials and services provided to meet the actual needs and demands of the affected population. This study sought to identify the perceptions of representatives of the international organizations providing this aid, the international workers involved with the delivery of this aid, the workers who were employed locally by the international organizations, the recipients, and the local authorities. This study sought to identify the perceptions of these personnel relative to the adequacies of the supplies in meeting the needs and demands of the population during and following the war in Bosnia-Herzegovina. METHODS: Structured interviews were conducted with representatives of international organizations and workers providing aid and with locally employed workers, recipients of the assistance, and the authorities of the areas involved. Descriptive and inferential statistics were used to assist in the analysis of the data. RESULTS: Eighty-eight interviews were conducted. A total of 246 organizations were identified as providing assistance within the area, and 54% were involved with health-related activities including: 1) the provision of medications; 2) public health measures; and 3) medical equipment or parts for the same. Internationals believed that a higher proportion of the needs were being met by the assistance (73.4 +/- 16.4%) than did the nationals (52.1 +/- 23.3%; p < 0.001). All groups believed that approximately 50% of the demands of the affected population were being addressed. However, 87% of the international interviewees believed that the affected population was requesting more than it actually needed. While 27% of the international participants believed that > or = 25% of what was provided was unusable, 80% of the recipients felt that > or = 25% of the provisions were not usable. Whereas two-thirds of the international participants believed that > or = 25% of the demands for assistance by the affected community could not be justified, only 20% of the recipients and authorities believed > or = 25% of the demands were unjustified. CONCLUSIONS: Many organizations are involved in the provision of medical assistance during a disaster. However, international organizations and workers believe their efforts are more effective than do the recipients. PMID- 11066838 TI - Earthquake epidemiology: the 1994 Los Angeles Earthquake emergency department experience at a community hospital. AB - INTRODUCTION: To assess the volume of patients and the composition of their injuries and illnesses that presented to an emergency department (ED) close to the epicenter of an earthquake that occurred in a seismically prepared area. METHODS: A retrospective analysis of data abstracted from charts and ED logs for patient census and types of injuries and illnesses of the patients who presented in the ED of a community hospital before and after the earthquake (6.8 Richter scale) that occurred in 1994 in Los Angeles. Illnesses were classified as trauma- and non-trauma related. Data were compared with epidemiological profiles of earthquakes in seismically prepared and unprepared areas. RESULTS: A statistically significant increase in ED patient census over baseline lasted 11 days. There was a large increase in the number of traumatic injuries such as lacerations and orthopedic injuries during the first 48 hours. Beginning on the third day after the event, primary care conditions predominated. When the effects of the LA quake were compared with those of similar Richter magnitude and disruptive capability, the ED epidemiology profile was similar to those in seismically unprepared areas, except for the total number of casualties. CONCLUSION: The majority of patients with traumatic injuries presented within the first 48 hours. The increase relative to baseline lasted 11 days. Efforts to develop disaster response systems from resources outside the disaster-stricken area should focus on providing mostly primary care assistance. Communities in seismically prepared areas could require external medical assistance for their EDs for up to two weeks following the event. PMID- 11066839 TI - Emergency department organisation for disasters: a review of emergency department disaster plans in public hospitals of Singapore. AB - Disaster management plans of emergency departments (EDs) in four major public hospitals were reviewed. A comparison was made between these plans, and they were analyzed to gain an understanding of the differing objectives and doctrines behind the practices. These were summarized into five major management concepts, which are considered to be critical to the success of a disaster plan: 1) staff mobilization systems (cascading vs batch mobilization); 2) staff deployment systems; 3) team organization (surgeons vs residents); 4) area management (the role of the area manager); 5) casualty volume management (accommodation vs expansion vs extension concepts). The concepts derived should serve as a useful guide to the development of an ED disaster plan and potentially influence how new ED facilities could be planned. PMID- 11066840 TI - Hospital responses to acute-onset disasters: a review. AB - INTRODUCTION: Hospitals the world over have been involved in disasters, both internal and external. These two types of disasters are independent, but not mutually exclusive. Internal disasters are isolated to the hospital and occur more frequently than do external disasters. External disasters affect the community as well as the hospital. This paper first focuses on common problems encountered during acute-onset disasters, with regards to hospital operations and caring for victims. Specific injury patterns commonly seen during natural disasters are reviewed. Second, lessons learned from these common problems and their application to hospital disaster plans are reviewed. METHODS: An extensive review of the available literature was conducted using the computerized databases Medline and Healthstar from 1977 through March 1999. Articles were selected if they contained information pertaining to a hospital response to a disaster situation or data on specific disaster injury patterns. Selected articles were read, abstracted, analyzed, and compiled. RESULTS: Hospitals continually have difficulties and failures in several major areas of operation during a disaster. Common problem areas identified include communication and power failures, water shortage and contamination, physical damage, hazardous material exposure, unorganized evacuations, and resource allocation shortages. CONCLUSIONS: Lessons learned from past disaster-related operational failures are compiled and reviewed. The importance and types of disaster planning are reviewed. PMID- 11066841 TI - Prior planning to avoid responders becoming "victims" during disasters. AB - Prior planning to meet the physical and mental needs of medical and emergency services responder, is a practical measure to reduce staff stress. This has the potential to improve both the operational efficiency of a disaster response and reduce the incidence of post-traumatic stress disorders in responders. Research is needed to define which interventions provide the greatest benefits to local responders. PMID- 11066842 TI - Emerging organizational structures in the ambulance industry in the United States. AB - This analysis seeks to identify emerging forms of organizations in emergency medical services (EMS) in the United States, to provide examples of them, to relate them to changes in healthcare generally, and to apply a classification scheme. Public policy issues related to these new forms of organizations and lessons from other areas of the healthcare system are identified. Recent changes in the healthcare system in the United States have been marked by modifications in the structure of organizations that provide and pay for health services. New forms of organizations and alliances among existing organizations have emerged in an effort to improve the efficiency of the services provided and to improve organizations' market positions. Reflecting increased competition within EMS and the demands of the changing health-care delivery system, several types of organizations have begun to emerge in EMS that resemble those occurring in health care generally. These include forms of horizontal integration, such as consolidated ambulance services and various models of ambulance service networks; and forms of vertical integration, such as demand management programs and public private joint ventures. The ultimate end might be complete integration with a carve-out of all non-scheduled care. Although changes in EMS organizations result largely from marketplace decisions by sellers and purchasers, this does not mean that there is no public policy role. While new organizational forms may increase the ambulance industry's efficiency, public policy makers must be concerned about quality and access as well. Some policy responses will promote marketplace changes, others will accept them generally, but will seek to correct problems, and a third group will attempt to restrain the market. PMID- 11066843 TI - Indifference, apathy, and preparedness. PMID- 11066844 TI - Survey research in disaster public health. AB - INTRODUCTION: While much has been learned during the past three decades of research in the disaster field, there still are some major gaps in knowledge. The need for more and better research on the health aspects of disasters is especially noted. Often, research into the health aspects has been anecdotal in nature and suffers from poor documentation of human losses. However, there are valid research methodologies that can be adapted to better document losses, evaluate interventions, and set priorities for investments to reduce the burden on the health of the population caused by disasters. METHODS: A number of data sources are used to demonstrate the potential uses of surveys in disaster health. The majority of the examples reflect data collected by telephone interviews following earthquakes in California. RESULTS: By using comparable instruments, it is possible to track the changes in preparedness levels across time. Similarly, it is possible to compare injury rates or other health impacts across time, place, and disaster type. In addition, risk factors can be identified for health outcomes. For example, in the Northridge earthquake, those over age 60 years were three times more likely to be hospitalized or die as a result of injuries than were those aged 20-59 years. Interventions can be evaluated. Slightly less than half of the respondents of the El Nino study had heard messages about preparing for the on-coming weather and their preparedness levels were not significantly different from those who had not heard about preparing for the weather. CONCLUSION: Surveys are useful tools for identifying and evaluating the health impacts of disasters. PMID- 11066845 TI - Pneumothorax during CPR training: case report and review of the CPR literature. AB - Cardiopulmonary resuscitation is taught widely to both lay persons and health care workers. It is a challenging psychomotor skill. Concerns about its safety to the rescuer have centered around the risk of infectious disease exposure. A young nursing assistant developed a minimally symptomatic pneumothorax during CPR training. This case is the first reported example of this complication for a CPR trainee or provider. The literature is reviewed for complications for CPR provider and recipient and the relevant issues regarding the current status and future direction of this intervention. PMID- 11066846 TI - Principles of disaster management. Lesson 7: Management leadership styles and methods. AB - This lesson explores the use of different management leadership styles and methods that are applied to disaster management situations. Leadership and command are differentiated. Mechanisms that can be used to influence others developed include: 1) coercion; 2) reward; 3) position; 4) knowledge; and 5) admiration. Factors that affect leadership include: 1) individual characteristics; 2) competence; 3) experience; 4) self-confidence; 5) judgment; 6) decision-making; and 8) style. Experience and understanding the task are important factors for leadership. Four styles of leadership are developed: 1) directive; 2) supportive; 3) participative; and 4) achievement oriented. Application of each of these styles is discussed. The styles are discussed further as they relate to the various stages of a disaster. The effects of interpersonal relationships and the effects of the environment are stressed. Lastly, leadership does not just happen because a person is appointed as a manager--it must be earned. PMID- 11066847 TI - You say you want a revolution. PMID- 11066848 TI - A changing future. Alternative medicine and reimbursement trends. PMID- 11066849 TI - PTs top "most wanted" list. PMID- 11066850 TI - Learn our lessons well. PMID- 11066851 TI - Treading water. PMID- 11066852 TI - Water-proofing. Measuring aquatic therapy effectiveness. PMID- 11066853 TI - Switching gears. PMID- 11066854 TI - Effectively using ultrasound. PMID- 11066855 TI - The bottom line: HMO and rehabilitation. PMID- 11066856 TI - Diabetes mellitus. PMID- 11066857 TI - Managing the diabetic foot. PMID- 11066858 TI - Cutting costs in workers' compensation. PMID- 11066859 TI - Communication is key in case management. PMID- 11066860 TI - Focusing on research. PMID- 11066861 TI - Bariatric rehab. PMID- 11066862 TI - Leading the way. Rehab providers have to develop adequate outcomes measurement systems for tomorrow's health care. PMID- 11066863 TI - The quake that rocked rehabilitation, Part IV. Survival may be as simple as ABC. PMID- 11066864 TI - Excess therapy. The Office of the Inspector General reports medically unnecessary therapies at SNFs. PMID- 11066865 TI - The power of two. UCSF and Stanford merge to prepare for changing market forces, finding the partnership beneficial to both rehab departments. PMID- 11066866 TI - Awakening as a change process among women at risk for HIV who engage in survival sex. AB - Women who exchange sex for survival requirements risk exposure to the human immunodeficiency virus (HIV). This grounded theory study was conducted to better understand these women's concerns, attitudes, and behaviors related to HIV, with the goal of informing prevention research. Interviews with 11 women engaged in survival sex were analyzed. Women described sliding into survival sex as a result of economic crises. Survival sex exposed women to violence, drug use, sexually transmitted infections, and HIV. Mitigating these risks is a process of awakening in which women reconstruct risk and survival and make changes in their behavior. These findings highlight the complexity of the problem of survival sex and suggest interventions to help women protect themselves against HIV. PMID- 11066867 TI - Rethinking professional attitudes in mental health settings. AB - Given that the psychiatric nursing standards developed by the American Nurses Association strongly advocate the inclusion of families in treatment settings, the study discussed in this article sought to determine how families were actually experiencing care in mental health settings for children and adolescents. The aim was to elicit the perspectives of parents whose children were hospitalized in psychiatric hospitals and to examine those experiences against the ideal rhetoric set forth by the standards of care. Results of in depth interviews with a purposeful sample of parents demonstrate that there is a significant gap between standards and practice. PMID- 11066868 TI - Core issues for female child abuse survivors in recovery from substance misuse. AB - Female survivors of childhood abuse have increased risk for substance misuse. Childhood abuse survivors describe current mental health and substance misuse services as not addressing central problems, the "heart of the matter." Negative core issues related to abuse events, contexts, substance use, and current problems were explored in open-ended interviews with 20 female abuse survivors. Most were marginalized because of cocaine use, poverty, and ethnic status. Narrative analysis involved within- and between-account comparisons through adequate paraphrasing of in vivo descriptions. Results include in-depth description of basic core beliefs, developmental core influences, and current core experiences. Implications for practice and policy include appreciation of the influence of negative core experiences on mental health, substance misuse recovery, and preparedness for adulthood demands. PMID- 11066869 TI - Protease inhibitors: changing the way AIDS case management does business. AB - The purpose of the qualitative evaluation study discussed in this article was to examine the AIDS case management model under which five nonprofit AIDS service organizations (ASOs) in Midcity were operating. The study was organized around 40 qualitative interviews with executive directors, directors, and case managers. The finding was that AIDS case management is evolving to accommodate the changing environmental/contextual conditions that have resulted from combination drug therapies (protease inhibitors) introduced in 1996. The agencies are responding to the changes individually rather than as a network, and responses vary among the agencies. Institutional theory, an examination of the interconnectedness of clients, the ASOs, and their environmental context guided the analysis of the findings. PMID- 11066870 TI - Rebuilding life: the experience of living with AIDS after facing imminent death. AB - The study described in this article explored how 7 men who believed they were dying from an AIDS-related illness began the process of rebuilding their lives following a course of successful combination therapy. Using constant comparison within and across interviews, three major themes emerged. Back to living was the realization they were going to live. Realizing my life has changed was associated with living with uncertainty, appearing healthy while living with AIDS, and letting go of their preoccupation with physical well-being. Readusting my life was associated with a change in daily routines, reevaluation of their sense of self, questioning their relationships, reengaging with society, and developing new life plans. Findings suggest that these participants felt living had become more difficult. PMID- 11066871 TI - Living with change: elderly women's perceptions of having a myocardial infarction. AB - The purpose of the study described in this article was to explore and describe elderly (70+ years) women's perceptions of having a myocardial infarction (MI). Structured and unstructured, open-ended, face-to-face interviews with 11 women were used to collect qualitative data. The central theme that emerged was living with change. Five phases were revealed: searching for a diagnosis, being hit with the reality, discovering the nature of the change, adjusting to the change, and moving on with the change. Throughout these phases, the women were faced with the challenges of being in control, managing uncertainty, making sense, being independent, and sheltering others. The continuous process of change in their lives was taken for granted by these women. By having an understanding of the perspective of elderly women who have an MI, nurses will be more effective when caring for these individuals. PMID- 11066872 TI - Effective and ineffective management of incontinence: issues around illness trajectory and health care. AB - In this article, findings from a study that investigated the impact of incontinence on individuals and the impact of effective and ineffective health interventions for the management of incontinence on patient careers and health care are reported. Twenty-seven participants whose incontinence was judged by health professionals as being successfully managed or unsuccessfully managed were interviewed. The participants were 19 women and 8 men (mean age: 61 years). Twenty-two participants experienced urinary incontinence, 4 people experienced both urinary and fecal incontinence, and 1 woman experienced fecal constipation. The findings have been set in the context of the management of chronic conditions and provide illuminating evidence that may be useful when reviewing health care and health services. People who were actively involved with their care and who were involved with decision making felt that their incontinence was being effectively managed. Multiple referrals and the marshaling and targeting of health professionals and services also appeared to be associated with effective management of incontinence. PMID- 11066873 TI - Reflections on "making somebody angry". AB - This article presents an account of the methodological and ethical issues associated with a critical ethnographic study that focused on the problems of providing nondiscriminatory health care to people with HIV/AIDS. The focus is on the debates that followed the publication of the final report. The debates were centered around (a) formal objections by the chair of the institutional ethics committee to the standard procedure of a discussion of the methodological limitations imposed by the requirements of the conditions of access; (b) reactions by a small section of the medical profession to adverse findings, which were published in a professional medical journal and included an attack on both the methods used and the ethics of the researchers; and (c) the response of the media, which selectively sensationalized and distorted the findings. This discussion may provide further insight into the problems of gaining access to clinical settings for the purpose of conducting qualitative research. PMID- 11066874 TI - Avoiding common pitfalls in qualitative data collection and transcription. AB - The subjective nature of qualitative research necessitates scrupulous scientific methods to ensure valid results. Although qualitative methods such as grounded theory, phenomenology, and ethnography yield rich data, consumers of research need to be able to trust the findings reported in such studies. Researchers are responsible for establishing the trustworthiness of qualitative research through a variety of ways. Specific challenges faced in the field can seriously threaten the dependability of the data. However, by minimizing potential errors that can occur when doing fieldwork, researchers can increase the trustworthiness of the study. The purpose of this article is to present three of the pitfalls that can occur in qualitative research during data collection and transcription: equipment failure, environmental hazards, and transcription errors. Specific strategies to minimize the risk for avoidable errors will be discussed. PMID- 11066875 TI - Scientific commentary: the scientific foundations and medical and social prospects of the Human Genome Project. AB - Author examines the scientific foundations of the genetics revolution and discusses the Human Genome Project, including its medical and social consequences. PMID- 11066876 TI - Genetic discrimination in the workplace. AB - Author argues that the Americans with Disabilities Act prohibits discrimination against workers based on their genetic makeup. He also examines state legislation and recently proposed federal legislation prohibiting genetic discrimination. PMID- 11066877 TI - Genetic privacy and confidentiality: why they are so hard to protect. AB - Author notes that widespread concerns have been raised about protecting genetic privacy and confidentiality in insurance and employment. He argues that effective protections are difficult because complicated issues, such as the right of access to health care, are invariably implicated. PMID- 11066878 TI - Reconsidering genetic antidiscrimination legislation. AB - In response to the awareness that genetic discrimination is a significant problem, many states have passed or are passing legislation regulating the use of genetic information. Authors discuss the weaknesses as well as the strengths of such legislation, and recommend that the laws be redrafted to prohibit discrimination on the basis of any type of predictive medical information. PMID- 11066879 TI - Oocyte cytoplasm transfers and the ethics of germ-line intervention. AB - Author advances the discussion about positive genetic selection by looking at the use of oocyte cytoplasm transfers as a possible means to prevent disease and to overcome fertility, focusing on the issues of family and kinship, commodification of eggs and egg donors, and germ-line alterations. PMID- 11066880 TI - Clinical commentary: the challenges of genetic medicine to the patient-physician relationship. AB - Author explores the interface between the current clinical reality for patients and evolving genetic research results, noting how knowledge about various genetic disorders is posing a major challenge to the patient-physician relationship. PMID- 11066881 TI - The Supreme Court confronts HIV: reflections on Bragdon v. Abbott. AB - Author examines in depth the U.S. Supreme Court's 1998 decision in Bragdon v. Abbott concerning the application of the Americans with Disabilities Act to HIV, and explains why the ruling was even necessary and what the decision says about the relationship between law and public health. PMID- 11066882 TI - Hospital consent for disclosure of medical records. AB - Disclosure of information clauses in general consent-to-treatment forms used by 202 large hospitals nationwide are described and mapped into a taxonomy to distinguish types of disclosures and to help structure consent documents. PMID- 11066883 TI - Managed care: Texas's Health Care Liability Act held partially preempted by ERISA. PMID- 11066884 TI - Antitrust: Fifth Circuit allows private benefit under state action doctrine. PMID- 11066885 TI - ERISA: state preferred provider statute preempted by ERISA. PMID- 11066886 TI - Malpractice: Alaska Supreme Court limits duty of hospitals to disclose risks of blood transfusions. PMID- 11066887 TI - Medicaid & Medicare: violations of health care laws found actionable under the FCA. PMID- 11066888 TI - Medicaid & Medicare: HCFA must monitor HMOs to ensure appeal rights for Medicare beneficiaries. PMID- 11066889 TI - Medicaid & Medicare: restrictions on Medicaid eligibility counseling found unconstitutional. PMID- 11066890 TI - Pain management and palliative care in the era of managed care: issues for health insurers. AB - Author reports on empirical study of medical directors at Blue Cross Blue Shield Plans regarding their awareness of and response to issues of pain management and palliative care for their insured populations. PMID- 11066891 TI - Public financing of pain management: leaky umbrellas and ragged safety nets. AB - Although many people in pain depend on public health care programs for aid, these programs cover pain relief only fragmentarily. He examines the gaps and deficiencies in Medicare and Medicaid funding of pain relief, and explores the effects of Medicare and Medicaid fraud enforcement on pain management. PMID- 11066892 TI - Criminal act or palliative care? Prosecutions involving the care of the dying. AB - Author examines criminal investigations and prosecutions of physicians and nurses in connection with their care of dying patients and concludes that the criminal law has failed to protect patients and families and has significant power to deter appropriate pain management for dying patients. PMID- 11066893 TI - In search of a new ethic for treating patients with chronic pain: what can medical boards do? AB - Author argues that a complex "ethic of underprescribing" underlies the continued reluctance of physicians to use opioids to treat chronic pain. She contends that state medical boards are uniquely positioned to promote a new ethic for pain management, but stresses the difficulties for boards in attaining this goal. She thinks success may hinge on whether boards can change their approach to pain management and persuade a skeptical medical community that adopting a risk for underprescribing will serve the long-term interests of patients and the profession. PMID- 11066894 TI - Commentary: the potential for unintended consequences from public policy shifts in the treatment of pain. AB - Authors caution against possible unintended consequences of intractable pain treatment acts, suggesting that health care professionals look to the guidelines prepared by the Federation of State Medical Boards for an approach to this issue. PMID- 11066895 TI - AIDS & HIV: Colorado court upholds privacy rights in disclosure of test results. PMID- 11066896 TI - Consent: Ohio Appellate Court affirms confidentiality claim. PMID- 11066897 TI - Consent: Pennsylvania Court expands scope of consent requirement. PMID- 11066898 TI - Contracts: district court holds FEHBA preempted by Pennsylvania statute. PMID- 11066899 TI - Disability & ADA: Third Circuit broadens scope of "qualified individual" under the ADA. PMID- 11066900 TI - EMTALA: Louisiana District Court limits scope of preemption. PMID- 11066901 TI - Fraud & abuse: Eighth Circuit rules that states are "persons" under the False Claims Act. PMID- 11066902 TI - Malpractice & negligence: Arizona Court affirms immunity of organ donation personnel. PMID- 11066903 TI - Mathematical formula for Swartz' Entropy Test Statistic. PMID- 11066904 TI - This CEO loves philanthropy! How a leader's support can raise a foundation's profile--and level of support. PMID- 11066905 TI - The win-win of charitable gift annuities. How to add this exciting option to your menu of deferred gift agreements. PMID- 11066906 TI - How to be the best possible personal solicitor. 10 keys to successful face-to face fund raising. PMID- 11066907 TI - Exploding the mountain. Philanthropy as a tool for change. PMID- 11066908 TI - Beyond cost-benefit analysis. Five steps to demonstrating community benefits. PMID- 11066909 TI - Patient confidentiality standards. AHP takes a strong stand. Association for Healthcare Philanthropy. PMID- 11066910 TI - Donor intent is not negotiable. Are we really good stewards? PMID- 11066911 TI - Building strong relationships with your board of trustees. Finding, keeping, and developing these key volunteers. PMID- 11066912 TI - No need for pain. AB - In an attempt to promote as pain free an experience as possible for patients and to improve how pain is managed in all settings, this 500-bed teaching hospital embarked on a hospital-wide quality improvement (QI) initiative. Initial measurement of clinicians' knowledge and attitudes related to pain and surveys on patient satisfaction identified improvement opportunities. Highlights of these findings and the major interventions taken are described. Education of all clinical staff on the major principles of pain management became the primary focus. Major outcomes realized to date have been: (a) a significant increase (p = .01) in physicians and nurses discussing with their patients the importance of managing their pain, (b) development of processes for documenting pain as the fifth vital sign, and (c) a 26% decrease in the use of meperidine. The next steps will be comprehensive monitoring of the effectiveness of pain management by clinical service, increased patient and family education on pain, and development of a formalized pain management competency program for clinical staff. PMID- 11066913 TI - Gregg O. Lehman on healthcare business coalitions, purchasing, and health policy. Interview by Joann Genovich-Richards. AB - As President and CEO of the National Business Coalition on Health (NBCH), Gregg Lehman leads a movement of 90 business coalitions nationwide seeking cost effective, better quality healthcare for employees and their families. Member coalitions represent more than 8,000 employers with more than 32 million employees and dependents. Dr. Lehman has 25 years of leadership experience in higher education, private business, and a national association. In his current position, Dr. Lehman is actively working with coalitions to promote their role in relation to value-based healthcare purchasing and health policy issues. In addition, he is actively developing NBCH into an enterprise that assists local coalitions in developing national contracts and strategic partnerships for healthcare products and services. Dr. Lehman earned a PhD in higher education administration, with a minor in finance and economics, from Purdue University. PMID- 11066914 TI - Multidisciplinary shared leadership. AB - Shared Governance evolved as a way for hospital nurses to have a role in decision making that affected nursing practice. This article describes how a multidisciplinary shared leadership program can be implemented in a hospital setting in order to empower all staff to participate in decision making and to continue the evolutionary process of continuous quality improvement. Various shared leadership models are described as well as a step-by-step implementation process and one hospital's story of the successful implementation of a council or model. PMID- 11066915 TI - Error-free healthcare: mission possible! PMID- 11066916 TI - Development and dissemination of minimum standards of care for asthma. AB - Asthma is a common illness in children and adults that is often associated with suboptimal outcomes, despite the existence and distribution of carefully considered national and international guidelines for diagnosis and treatment. The need to improve asthma care in North Texas motivated a coalition of health plans, employers, hospitals, academic medical centers, providers, and benefit consultants to collaborate in developing and implementing Minimum Standards of Care for Asthma. To gain consensus, the North Central Texas HEDIS Coalition used a panel of regional asthma experts and surveyed a large number of primary-care providers to construct a one-page document that was ultimately approved by regional stakeholders in healthcare participating in this coalition and also by specialty organizations in Texas. Adopting a minimum standard guideline, developed and supported by multiple stakeholders, may represent a more realistic intermediate goal when implementing "best practice" guidelines is limited by practical barriers. PMID- 11066917 TI - CONQUEST 2.0: an emerging clinical performance measurement tool. AB - Variations in quality of healthcare have existed for many years, yet interest in measuring, understanding, and eliminating these variations has waxed and waned. The advent of managed care and its perceived focus on reducing costs has stimulated interest in variations in quality of care. In response to the need to better understand and measure quality, the Agency for Health Care Policy and Research (AHCPR) funded the development of a tool capable of categorizing clinical performance measures, a subset of quality measures, for comparing and evaluating them relative to a user's specific needs for measuring and improving quality. This research lead to the development of the COmputerized Needs-oriented QUality measurement Evaluation SysTem (CONQUEST)--a free software quality improvement tool that includes about 1,200 clinical performance measures. CONQUEST enables users to quickly identify measures of interest, compare them on the basis of factors of importance, and select measures that will help them measure and improve care. PMID- 11066918 TI - The story of the rebuilding of a team. AB - There once was a team that dreamed of being highly functional. Team members believed they knew the path--they knew their expected results, their goals, and their decision-making authority. Yet, the team was miserable in its work. In order to find out what was wrong, the team contacted the team expert in the organization whose job was to build teams. The team expert determined that the process components of personal interaction criteria were being overlooked and, instead, content work was being pursued. As a result of these findings, the team rebuilt itself by defining its purpose in terms of goals and objectives, determining a set of team values and behaviors, and setting ground rules for meeting management. PMID- 11066919 TI - Monitoring steam sterilization practices in primary care settings. AB - Steam sterilization is a common method for processing reusable instruments in ambulatory care settings. Ensuring that sterilization practices are consistent with accepted standards is a quality-of-care parameter as well as an infection control issue. Factors that must be considered in improving performance in this important area are: operator training and continued competency, selection of chemical and biological media and methods, work flow controls, documentation methods and standards, written policies and procedures to support practices, and ongoing monitoring of the system and practices with feedback to responsible persons. PMID- 11066920 TI - Focus on voice recognition. Talking up an emerging technology. PMID- 11066921 TI - Focus on voice recognition. Technology moves into the mainstream. PMID- 11066922 TI - Making software upgrades a first-class experience. PMID- 11066923 TI - Internet pharmacies: all hype with no help? PMID- 11066924 TI - HIPAA and health information privacy rules: almost there. PMID- 11066925 TI - A Blue's approach to disease management. PMID- 11066926 TI - Disease management: a 'smart' way to interact with patients. PMID- 11066927 TI - Nursing technology ...@ the speed of thought? PMID- 11066928 TI - Merging healthcare and the Internet in the new century. PMID- 11066929 TI - Hotlist. Managed care. PMID- 11066930 TI - What works. Viewer gets Center out from under paperwork.. PMID- 11066931 TI - What works. Timing is everything in the OR. PMID- 11066932 TI - What works. Old system gets modern touch. PMID- 11066933 TI - Healthcare's evolution in the technological universe. PMID- 11066934 TI - Industry expected to seek major changes, delay in revised PPS. PMID- 11066935 TI - OIG audit of PPS claims reveals improper payments to Medicare Part B suppliers. PMID- 11066936 TI - Labor data shows SNF employee injury rates decline; musculoskeletal disorders, ergonomics regulation still hotly debated. PMID- 11066937 TI - National nursing facility survey provides details on industry. PMID- 11066938 TI - Get more out of your medical management efforts. PMID- 11066939 TI - Efficient medical management begins with adoption of rigorous clinical guidelines. PMID- 11066940 TI - Tightly drawn contracts can make or break specialty carve-outs. PMID- 11066941 TI - Wound care contract represents new wrinkle for risk agreements. PMID- 11066942 TI - Move now to align physician compensation with market. PMID- 11066943 TI - Group blends two income streams into physician compensation formula--and survives. PMID- 11066944 TI - Use this checklist to assess your readiness for risk. PMID- 11066946 TI - Can a capitated hospitalist program survive in a FFS world? PMID- 11066945 TI - Use these data to maximize your contribution margins. PMID- 11066947 TI - Priced out. PMID- 11066948 TI - Championship management for healthcare organizations. AB - Stakeholders will put increasing pressure on integrated health systems (IHS) for measured performance, demanding data on quality and patient satisfaction, while simultaneously pressing for lower cost. The changes to Joint Commission on Accreditation of Healthcare Organizations (Joint Commission) and the growing importance of the National Committee on Quality Assurance (NCQA) are simply forerunners of an intensifying trend. Quality of care in particular will face increasing scrutiny. Achieving competitive targets in these areas will also require measures addressing demand and worker satisfaction. "Balanced scorecard" approaches will allow IHS and their accountable work groups to track performance on several dimensions and establish integrated goals or targets. Those with consistently good scores will be labeled "champions." Champions will support the multidimensional measures with improved decision processes. About eight major processes will be central--governance/strategic management, clinical quality, clinical organization, financial planning, planning and marketing, information services, human resources, and plant services. It is possible to map these processes to the criteria of the Joint Commission, NCQA, and Malcolm Baldrige Quality Award. The processes themselves can be measured and common weaknesses identified and corrected. Champions share some common characteristics that seem to arise from the combination of processes and measures. Among these characteristics are service line orientation, extensive partnering with other organizations, and the possibility of outsourcing organizational components. PMID- 11066949 TI - Structuring managed care: lessons from traditional insurance. PMID- 11066950 TI - Thomas C. Dolan, Ph.D., FACHE, CAE, president and chief executive officer of the American College of Healthcare Executives. Interview by James A. Johnson. PMID- 11066951 TI - Ethical guidance in the era of managed care: an analysis of the American College of Healthcare Executives' Code of Ethics. AB - Market competition and the rise of managed care are transforming the healthcare system from a physician-dominated cottage industry into a manager-dominated corporate enterprise. The managed care revolution is also undermining the safe guards offered by medical ethics and raising serious public concerns. These trends highlight the growing importance of ethical standards for managers. The most comprehensive ethical guidance for health service managers is contained in the American College of Healthcare Executives' (ACHE) Code of Ethics. An analysis of the ACHE Code suggests that it does not adequately address several ethical concerns associated with managed care. The ACHE may wish to develop a supplemental statement regarding ethical issues in managed care. A supplemental statement that provides more specific guidance in the areas of financial incentives to reduce utilization, social mission, consumer/patient information, and the health service manager's responsibility to patients could be extremely valuable in today's complex and rapidly changing environment. More specific ethical guidelines would not ensure individual or organizational compliance. However, they would provide professional standards that could guide decision making and help managers evaluate performance in managed care settings. PMID- 11066952 TI - The future of employment-based health insurance. AB - A transformation of employment-connected health insurance from a defined benefit to defined contribution arrangement is projected based on new economic realities affecting the competitiveness of the business environment. This article discusses those new realities along with the future of employment-based health insurance. The business of American business is profits, but, to the detriment of that goal, for the past half century business has also been in the business of providing health insurance for workers. However, in light of previously unencountered pressures on profits, employers are realizing they cannot afford to continue the practice of paying for and overseeing the provision of healthcare benefits to employees amid increasing premiums, state and federal mandates, the overbearing cost of managing healthcare benefits, and the threat of loss of protection under ERISA. Yet, the political and social pressures on businesses to continue to provide health insurance are formidable, perhaps impregnable, barriers to complete withdrawal of what has come to be thought of as a "right" of employees. Companies are anxious to find alternatives to the status quo, but any feasible alternative must cost less, require less administrative oversight, and ensure that employees still maintain a measure of choice. Two possible solutions for American businesses are adoption of (1) a "medical savings account" system, or (2) a "voucher" system. Either system would result in lower costs and greater fiscal stability for both employers and employees. They would also remove much of the responsibility for healthcare decisions from employers and place it in the hands of the employees. But, perhaps the greatest contribution of either system would be the reduction in moral hazard and its inflationary effect on medical costs. PMID- 11066953 TI - Strategic performance management: development of a performance measurement system at the Mayo Clinic. AB - Managing and measuring performance become exceedingly complex as healthcare institutions evolve into integrated health systems comprised of hospitals, outpatient clinics and surgery centers, nursing homes, and home health services. Leaders of integrated health systems need to develop a methodology and system that align organizational strategies with performance measurement and management. To meet this end, multiple healthcare organizations embrace the performance indicators reporting system known as a "balanced scorecard" or a "dashboard report." This discrete set of macrolevel indicators gives senior management a fast but comprehensive glimpse of the organization's performance in meeting its quality, operational, and financial goals. The leadership of outpatient operations for Mayo Clinic in Rochester, Minnesota built on this concept by creating a performance management and measurement system that monitors and reports how well the organization achieves its performance goals. Internal stakeholders identified metrics to measure performance in each key category. Through these metrics, the organization links Mayo Clinic's vision, primary value, core principles, and day-to-day operations by monitoring key performance indicators on a weekly, monthly, or quarterly basis. PMID- 11066954 TI - Laying the foundation for successful physician-health system partnerships. PMID- 11066955 TI - From advocacy to ambassadorship: physician participation in healthcare governance. AB - Increasingly, physician leaders are called upon to assume governance positions in healthcare organizations. These range from small group practices to integrated systems with a regional or even national presence. Yet the meaning of board membership and the skills required for effective contributions in the boardroom are often unclear. This article reviews the essential challenges to governance bodies in the healthcare field, and focuses particularly on the issue of ambassadorship--an often neglected but essential aspect of effective board participation. PMID- 11066956 TI - Nontraditional services provided by nonprofit and for-profit hospitals: implications for community health. AB - In part because of reimbursement changes in the 1980s, hospitals became involved in health promotion and disease prevention activities often to attract patients. Today, these services may have an effect on the burden of disease and on illness prevention in some communities. Given the changes anticipated in healthcare delivery, assessing the scope of these services and integrating them with other private-public efforts is of utmost importance. Here we use a 1993 survey of all 4,977 private medical and surgical hospitals in the United States to determine the scope of disease prevention, health enhancement, and palliative services provided by facility type, geographic location, and institutional ownership. We found that church-operated and other nonprofit hospitals appear to provide a spectrum of palliative and preventive health services both for their patients and those in the local community. Given their apparent scope, these services could have an effect on the burden of disease and on illness prevention in many communities. With major changes anticipated in future healthcare delivery and the recent failures reported for many community health intervention programs, healthcare administrators need to focus on ways to integrate their services with other private and public health efforts. If this could be achieved, then private hospitals could be more successful in serving their local communities and in enhancing the public's health in the new century. This article outlines several basic steps to assist administrators in achieving these goals. PMID- 11066957 TI - Interview with Graham T.T. Molitor, president of Public Policy Forecasting and vice president of the World Future Society. Interview by James A. Johnson. PMID- 11066958 TI - Economic models for physician-health system partnerships. PMID- 11066959 TI - Nothing but Net. PMID- 11066960 TI - Medicare and the rules of national policymaking: if A, then B. PMID- 11066961 TI - Creating a healing environment: the importance of the service setting in the new consumer-oriented healthcare system. AB - Over the last ten years, the healthcare industry has recognized that the physical environment is a valuable resource that can and does affect all of its customers. Although most service organizations give some thought to setting, its importance to the service experience has been most thoroughly understood by those who view and treat their customers as guests, that is, the guest service industry. An excellent healing environment will reinforce excellent clinical quality, but an inferior environment can detract from fine clinical care. One of the most important principles learned by the guest service industry is to provide the setting customers expect. Another is to create an environment that meets or exceeds customer needs for safety, security, support, competence, physical comfort, and psychological comfort. This article provides a detailed discussion of how such an environment can be created in healthcare facilities drawing from the experience of the best guest service organizations. PMID- 11066962 TI - Interview with Jeptha Dalston, Ph.D., President and CEO of AUPHA and ACEHSA, Washington, D.C. Interview by Helen-Joy Bechtle. PMID- 11066963 TI - Other-than-economic models for physician-health system partnerships. PMID- 11066964 TI - Leaner is greener. PMID- 11066965 TI - Loyalty in managed care: a leadership system. AB - Healthcare executives are given a comprehensive and integrated ten-step system to lead their organization toward stabilizing a financial base, improving profitability, and differentiating themselves in the marketplace. This executive guide to implementing loyalty-based leadership can be adapted and used on an immediate basis by healthcare leaders. This article is a useful resource for healthcare executives as they move to make loyalty an organizational resource. Effectively managing the often-fragmented forces of loyalty can produce a healthier bottom line and improve the commitment among key stakeholders within a managed care environment. A brief loyalty-based leadership practices survey is included to serve as a catalyst for leaders and their teams to strategically discuss loyalty and retention in their organization. PMID- 11066966 TI - Capitated contracting roles and relationships in healthcare. AB - Capitated contracting of health providers has created substantial change in healthcare markets. This article assesses how capitation affects the roles and relationships of healthcare organizations. In-depth case studies were conducted of eight major hospital-led integrated health networks/systems and two large integrated medical groups. Types of capitated contracts employed, contract support capabilities developed, relationships among providers in the support services, and lessons learned about capitation were explored. The experiences of these organizations provide valuable guidance for health executives as they develop or refine capitated contracting strategies. PMID- 11066967 TI - Building partnerships with the community: lessons from the Camden Health Improvement Learning Collaborative. AB - This case study describes the Camden City Health Improvement Learning Collaborative (the Collaborative), a community care network initiative formed in 1993. The organization is composed of representatives from local healthcare providers, public agencies, religious organizations, and neighborhoods. The major goal of this initiative is to improve the health status of the community by involving and empowering residents in the solution of their needs. The Collaborative represents a grassroots strategic model of community inclusion in the formulation of goals and programs to improve community health status. The case study describes the dynamics of the Collaborative by examining the following: historical development; political, institutional, and social context; planning process; organization and structure; and performance evaluation. The article concludes with a discussion of the strategic and operational lessons learned from the Collaborative. PMID- 11066968 TI - E-health: reinventing healthcare in the information age. PMID- 11066969 TI - Analysis of Blues' costs offers fascinating insights. PMID- 11066971 TI - Assessing the value equation can help guide drug usage. PMID- 11066970 TI - Cost not the primary factor driving drug coverage. PMID- 11066972 TI - Cost, utilization data focus on organ and tissue transplants. PMID- 11066973 TI - Do nurse practitioners deliver cost-effective care? PMID- 11066974 TI - How will PPS realign financial incentives and care? An indepth panel discussion by NAHC experts. National Association for Home Care. AB - The new prospective payment system (PPS) that will take effect for Medicare participating agencies on October 1, 2000 will create a dramatic change in the culture of home care that has existed since the inception of the home care benefit in 1965. Will these changes be for the better or for the worse? Who will be the winners, and who will be the losers? How will these incentives change the behavior of home care agencies and the caregivers they employ in their efforts to provide health care to the infirm and disabled? PMID- 11066975 TI - National Association for Home Care Code of Ethics. PMID- 11066976 TI - Balancing books & balancing values: the hidden threat of PPS. AB - Losing sight of the real vision and values of home care is a threat that becomes more real as agency leaders try to redefine their agencies to be PPS ready. Agencies must remain committed to ensuring that the foundation of the organization is one of sound business practices coupled with solid core values. PMID- 11066977 TI - Creating a forum for ethical decisionmaking. AB - The Central Florida Visiting Nurse Association created a multidisciplinary forum to help home care clinicians address tough ethical issues and other topics of concern. One tool that this group created, the Patient Care Contract, is a primary factor in this forum's success. PMID- 11066978 TI - HCFA responds to HAA questions. Hospice Association of America. PMID- 11066979 TI - Home Care Aide Association of America Code of ethics. PMID- 11066980 TI - OASIS inter-rater reliability. PMID- 11066981 TI - Media join call for Medicare home health relief. PMID- 11066982 TI - Ethics in home care agencies. AB - Ethics committees offer the opportunity to air ethical differences in a nonadversarial manner, enabling those involved to discuss and understand their differences. Establishing processes for an ethics committee involves reflection on basic ethical principles of conduct governing an individual or group. Agencies can learn from the process of ethical decisionmaking in home care organizations that have achieved accreditation either through CHAP or through JCAHO. PMID- 11066983 TI - Educating our future leaders. PMID- 11066984 TI - Emotions at work. PMID- 11066985 TI - Retaining current leaders. A gold mine in your back yard. PMID- 11066986 TI - Cultivating leaders with an acceleration pool. PMID- 11066987 TI - The lack of diversity at the top. PMID- 11066988 TI - Focusing on customer service. PMID- 11066989 TI - Unlearning lousy advice. PMID- 11066990 TI - www.healthcarefuture.com. PMID- 11066991 TI - Customer service will distinguish hospitals of the 21st century. PMID- 11066992 TI - Participants ask how to ensure the quality of care into the future. PMID- 11066993 TI - Technological innovation is one of many keys to the physician relationship. PMID- 11066994 TI - Summit speakers and participants define a good leader. PMID- 11066995 TI - Making choices. Elective procedures. PMID- 11066996 TI - Building and funding hospitals the non-traditional way. PMID- 11066998 TI - The Assessment Centre--Halton. PMID- 11066997 TI - Activity database. PMID- 11066999 TI - Facility management in German hospitals. AB - Facility management and optimum building management offer for hospitals a chance to reduce costs and to increase quality, process sequences, employee motivation and customer satisfaction. Some years ago simple services such as cleaning, catering or laundry were outsourced. Now, German hospitals progress to more complex fields such as building and medical technology, clinical support processes such as pharmacy, central laboratory and sterilization, goods and logistics services. PMID- 11067000 TI - Integrated facilities management information system. PMID- 11067001 TI - Ensuring ethical governance in your organization. PMID- 11067002 TI - Have physicians taken control of managed care? PMID- 11067003 TI - Designing the IDS business portfolio. AB - The business-value model is a strategic and business decision-making tool that can help integrated delivery systems (IDSs) evaluate the relative value that their individual businesses contribute to their overall systems. To use the model, IDSs first need to identify their individual business lines using criteria such as scope, size, marketability, operations, and competition. Businesses then are categorized as core, continuum, cash, or complementary based on four criteria: system purpose, size as reflected by annual revenue, competitive position, and level of profitability. Standards for these criteria should be set by the IDS in accordance with its market circumstances. PMID- 11067004 TI - Pain management improves care and revenue: an interview with ProCare Systems. AB - As provider and managed care organizations continue to look for better ways to control costs and improve patient outcomes, disease management programs are getting an increasing share of their attention. One often-over-looked area with significant potential to improve outcomes, reduce costs, and enhance revenues is pain management. It has been estimated that at least 40 percent of senior citizens suffer from chronic pain, and as the population ages, the number of chronic pain sufferers will only increase. Pain management companies have been forming to meet the current and future demand for comprehensive pain management programs. One such company is ProCare Systems, a single-specialty physician practice management company based in Grand Rapids, Michigan. HFM spoke with Fred N. Davis, MD, president and cofounder of ProCare Systems, and Cyndy Walsh, ProCare System's CEO, about pain management programs and the patient care and financial impact they can effect. PMID- 11067005 TI - APCs: reimbursement implications. AB - A transition to a new Medicare outpatient prospective payment system (PPS) will begin in July 2000, affecting many of the outpatient services provided by hospitals that participate in the Medicare program. The outpatient PPS will rely on ambulatory payment classifications (APCs) to classify outpatient services. Hospitals should anticipate that discounts and other factors will affect APC payments and take into account the impact that the new system will have on their revenue. PMID- 11067006 TI - Ambulatory surgery: next-generation strategies for physicians and hospitals. AB - Physicians' interest in investing and practicing in independent ambulatory surgery centers (ASCs) has grown. In the face of physician involvement in ASC development, healthcare organizations must contend with possible loss of surgical volume and revenues as well as decreased physician support and loyalty to the organization. Healthcare organizations can encourage physicians to remain in the organization by addressing physicians' concerns about the financial prospects and efficiency of independent and hospital-based ambulatory surgical arenas. PMID- 11067007 TI - New ventures require accurate risk analyses and adjustments. AB - For new business ventures to succeed, healthcare executives need to conduct robust risk analyses and develop new approaches to balance risk and return. Risk analysis involves examination of objective risks and harder-to-quantify subjective risks. Mathematical principles applied to investment portfolios also can be applied to a portfolio of departments or strategic business units within an organization. The ideal business investment would have a high expected return and a low standard deviation. Nonetheless, both conservative and speculative strategies should be considered in determining an organization's optimal service line and helping the organization manage risk. PMID- 11067008 TI - Equity MSO (management services organization) benefits hospital and physicians. AB - Equity MSOs offer an alternative to hospital's employment of physicians and acquisition of their practices. In one example, several primary care physicians, organized into a single professional corporation, entered an agreement with a hospital to establish an equity MSO that was jointly owned by both parties. The partnership enabled the physicians to negotiate managed care contracts more skillfully and improve their bottom line. Under the agreement, the physicians' income is determined by their productivity. The hospital is confident that this new entity will help it sustain relationships with the physicians. PMID- 11067009 TI - Court affirms Medicare withholding of outlier underpayments. PMID- 11067010 TI - Skill-based pay improves PFS (patient financial services) staff recruitment, retention, and performance. PMID- 11067011 TI - Providers need e-mail security management. PMID- 11067012 TI - Making diversity a reality. PMID- 11067013 TI - Data trends. Why healthcare marriages end in divorce. PMID- 11067014 TI - U Mich: applying community oriented policing to healthcare settings. PMID- 11067016 TI - State association launches program to secure transmission of health data. PMID- 11067015 TI - Police/private security partnerships: how hospitals can participate. PMID- 11067017 TI - Hospital uses cameras to nab employee suspect in computer thefts. PMID- 11067018 TI - Two case histories: surviving and even benefiting from hospital security mergers. PMID- 11067019 TI - How one hospital tackles the drug theft/diversion problem head-on. PMID- 11067020 TI - Hospital's umbilical cord alarm fails to stop infant kidnapper. PMID- 11067021 TI - Medical center credits CCTV for 60% reduction in crime. PMID- 11067022 TI - What mock drills revealed about hospital's abduction response plan. PMID- 11067023 TI - The reality of violence in the workplace: stop being scared; start acting smart. AB - Violence in the workplace, including incidents associated with domestic abuse, has been causing increasing concern, and for good reason. Each year, more than two million people become victims of violent crime at work, according to the U.S. Bureau of Labor Statistics. A recent survey by Pinkerton Inc. of Fortune 1000 security executives revealed that workplace violence is considered the most important security threat to America's largest corporations. Based on these statistics and the rising number of incidents across the country, what should be done by various organizations in terms of intervention and prevention? What are the early clues and warning signs to possible threats of violence at the work site? And what is the role of the security director and others in making the workplace safe? First and foremost, three leading consultants on workplace violence recommend the formation of multidisciplinary threat assessment teams- supported by top management and including participation from security and area law enforcement--to develop violence prevention plans for increasing awareness and spelling out rules and procedures for heading off trouble. Other suggested steps to reduce the threat and likelihood of violence occurring also are presented in this report. PMID- 11067024 TI - The Colorado tour bus accident: emergency responses of area hospitals. PMID- 11067025 TI - Clean and sober. Residents turned a Victorian-era factory into Phoenix House's newest drug treatment facility. PMID- 11067026 TI - The new medical team: clinicians, technicians and ... patients? PMID- 11067027 TI - Ensuring a customer-focused experience: two success stories. PMID- 11067028 TI - The power of pharmacoeconomics. PMID- 11067029 TI - Keeping in touch in cyberspace. PMID- 11067030 TI - Evaluating your ethics committees. PMID- 11067031 TI - Physician practice management in the new millennium. PMID- 11067032 TI - Advice from the experts. PMID- 11067033 TI - Combining function and form. PMID- 11067034 TI - The long-term outlook for medical education. PMID- 11067035 TI - The consumer revolution: an age of changing expections. PMID- 11067036 TI - Cost-effectiveness analysis with defined budget: how to distribute resources for the prevention of cardiovascular disease? AB - The aim of the study was to undertake cost-effectiveness calculations subject to a defined budget. The setting chosen was the prevention of cardiovascular disease (CVD) by means of three intervention programmes in a Swedish county council. The population in the county was divided into subgroups according to risk level. For each subgroup the cost per years of life saved was calculated, as well as the annual budget claims. The budget available was defined as present direct cost in the programmes. The calculations resulted in a programming solution showing the optimal distribution of resources between the programmes. Also a league table was constructed and the cut-off value for a 'acceptable' cost-effectiveness was shown. The conclusion that can be drawn is that a combination of internationally published intervention results and local data regarding epidemiology and resource improves the accuracy and usefulness of cost-effectiveness ratios. However, the model presented is a first attempt containing only three interventions: the planned next phase is to integrate more interventions in the model. PMID- 11067037 TI - Supply, utilisation and outcome in hospital systems: an Anglo-Czech comparison. PMID- 11067038 TI - The effect of managed care on hospital staffing and technological diffusion. PMID- 11067039 TI - Can corporatization contribute to quality assurance and cost control in the German hospital sector? A pilot project for stem cell transplantation. PMID- 11067040 TI - Bigger bucks in '99--but balloon may burst. PMID- 11067041 TI - Crash course. How to build management skills--pronto. PMID- 11067042 TI - Cold war. EPA battles ozone depletion, tightens refrigerant regs. PMID- 11067043 TI - Face-off. Can your team take on the JCAHO crew? PMID- 11067044 TI - CHA moves forward on leadership development, BBA. PMID- 11067045 TI - Social accountability: the next generation. PMID- 11067046 TI - Compliance group posts best practices archive. PMID- 11067047 TI - Focus on goals, not technology. PMID- 11067048 TI - Scenario planning a useful tool in building strategic intent. PMID- 11067049 TI - Innovations in pain management. Catholic providers are in the forefront of treating the pain of chronic and terminal illness. PMID- 11067050 TI - Moving medical records online. "Community health record" puts needed information at providers' fingertips. PMID- 11067051 TI - When sponsors become partners. PMID- 11067052 TI - Catholic identity in a challenging environment. PMID- 11067053 TI - Structures in healthcare ministry in the church. PMID- 11067054 TI - Mission to Croatia. Franciscan team helps a new nation create its geriatric care system. PMID- 11067055 TI - Productivity is part of mission. The ministry needs to move past "ethereal discussions," a CEO says. PMID- 11067056 TI - Living our promises, acting on faith. CHA's benchmarking project will identify the core characteristics of Catholic healthcare. PMID- 11067057 TI - Mission-based investing comes of age. How two Catholic healthcare systems are making MBI (mission-based investing) a reality. PMID- 11067058 TI - Community networks. Partnerships between Catholic charities and Catholic healthcare organizations. Catholic Community Care, Cleveland. PMID- 11067059 TI - Employee education pays off for St. Joseph Health System. PMID- 11067060 TI - What Nancy taught me. PMID- 11067061 TI - The BBA's challenge to mission-driven hospitals. PMID- 11067062 TI - Doing things with--not to--the community. PMID- 11067063 TI - Using the Web to extend services. PMID- 11067064 TI - Discount store reborn as seniors' health center. PMID- 11067065 TI - Challenges and competencies. The theological and spiritual aspects of Catholic healthcare leadership. PMID- 11067066 TI - Evaluating a system chief executive. Eight questions trustees should ask themselves about CEOs and CEO candidates. PMID- 11067067 TI - Medical waste and healthcare ethics. PMID- 11067068 TI - Capturing the power of the proxy. Catholic healthcare organizations can use their investments to promote social justice. PMID- 11067069 TI - Spirituality and healing. PMID- 11067070 TI - When the community cares. PMID- 11067071 TI - End-of-life care and Catholic ethics. PMID- 11067072 TI - Organizational assessment of end-of-life care. PMID- 11067073 TI - St. Francis Hospice shares its message to build success. PMID- 11067075 TI - Sophisticated Web sites lend a competitive edge. PMID- 11067074 TI - Improving attitudes toward change. PMID- 11067076 TI - A spiritual role for the elderly. PMID- 11067077 TI - Group purchasing pays dividends. PMID- 11067078 TI - A clearing in the woods. Retreat house aids mission integration at Indianapolis hospital. PMID- 11067079 TI - Managing hope in the workplace. Five simple strategies can help transform organizations. PMID- 11067080 TI - A new paradigm for investment screening. Four principles can help make the screening process less painful. PMID- 11067081 TI - Holy memory, faithful action. The Catholic identity of Catholic Health East, based in memory, has important implications for the future. PMID- 11067082 TI - "Once more unto the breach!" Why we must renew the struggle for healthcare reform. PMID- 11067083 TI - Ending the chaos in our healthcare system. PMID- 11067084 TI - Reducing the number of uninsured. PMID- 11067085 TI - Healthcare reform and the "consistent ethic". PMID- 11067086 TI - Community networks. Partnerships between Catholic charities and Catholic healthcare organizations. Caritas Communities, Youngstown, OH. PMID- 11067087 TI - Innovative Easter Seal programs help the disabled. PMID- 11067088 TI - Redefining processes to make hospitals more humane. PMID- 11067089 TI - The American College of Medical Practice Executives' competency study. AB - This article is the first of two studies conducted by the American College of Medical Practice Executives (ACMPE) that examines the perceived roles of medical practice executives. (Founded in 1956, the American College of Medical Practice Executives is the professional development and credentialing arm of the Medical Group Management Association (MGMA)). This study asked groups of physicians and nonphysician administrators to identify the competencies and associated skills and knowledge for administering group practices in today's changing environment. Those surveyed included administrators who are Fellows in ACMPE and 795 physicians who comprise the Society of Physician Administrators of the Medical Group Management Association. The responses were examined through a framework provided by the Managed Care Process Model. In this model, the focus is on the administrative and clinical processes required by different levels of managed care market penetration. The model progresses from a focus on relatively traditional practice management functions to those activities that are more complex with a greater focus on the integration of both clinical and business processes aimed at the health of populations. The analysis of the perceived competencies indicated that while both executive types perceived the importance of managing the health of populations, that task is not yet being incorporated into their professional roles. PMID- 11067090 TI - Information technology and medical group management. AB - Information technology permits revised patient management activities for high quality, cost-effective care in ambulatory clinics. The electronic medical record is financially feasible for very small physician groups. Disease management for chronic and terminal care patients represents an expanding area of service in medical group management. The Internet provides access to health care information that has empowered patients and their families to approach patient-physician office visits from a new relationship. Data provided by this information technology permit benchmarking of activities by ambulatory care service, treatment modalities, specialty group, and physician. PMID- 11067091 TI - Using the data you already have. AB - Managing a group practice requires constant attention to financial and operational data to achieve a sustainable business concern. By identifying key performance areas, a group practice can gather, report, and analyze data that are readily available but often overlooked. Although performance indicators will not solve all management problems, the insight gleaned from historical and projected data trending, in addition to external comparison and benchmarking, can result in proactive analysis and problem identification, mitigating the troubling impacts of ignorance and surprise. PMID- 11067092 TI - Integrated risk assessment and feedback reporting for clinical decision making in a Medicare Risk plan. AB - The challenge of tapping into the rich resource of population-based, aggregated data to inform and guide clinical processes remains one of the largely unrealized potentials of managed care. This article describes a multifaceted approach of using health-related data to support providers in clinical decision making as an adjunct to case management and primary care delivery. The goal is to provide data that can be used for clinical decision making that is population based, yet individualized for specific patient care situations. Information reporting holds great potential in the clinical care of patients because it can be used to identify persons who could benefit from early detection, intervention, or treatment. It has been suggested that one of the keys to success in managed Medicare is the timely use of information that is detailed, comprehensive, and real-time describing key parameters of clinical encounters. PMID- 11067093 TI - Physician compensation: rewarding productivity of the knowledge worker. AB - Designing a physician incentive compensation plan that aligns the demands of managed care with the perceived fairness of income distribution is a key challenge for medical practices today. Rather than focus on traditional productivity measures, managed care requires physicians to demonstrate efficient practice of medicine. Physicians still need to be highly productive; however, they are now required to demonstrate efficiency related to clinical resource management, patient access and service, and evidence-based outcomes. Approaches to the development of physician incentive compensation plans and case examples are offered to assist practices that are transitioning physician compensation from volume-based to efficiency-based indicators. PMID- 11067094 TI - Physician productivity and quality--a commentary. PMID- 11067095 TI - Benchmarking medical group practices using claims data: methodological and practical problems. AB - As claims data for physicians and groups of physicians has improved in quality and quantity, health information vendors have begun marketing information about medical groups' productivity, utilization, and quality. Based on interviews with product developers and our understanding of the evolution of their products, several methodological and practical issues remain. For now and the immediate future, health information vendors will continue to face the limitations of physicians' claims data. Vendors and purchasers should be aware of common data shortcomings such as inadequate monthly enrollment figures, possible physician upcoding to circumvent utilization management restrictions, and incorrect coding when a test is used to rule out a disease. In the longer term, several avenues seem likely to make medical groups' data better and richer because of computer based medical records and efficiencies possible from the Internet. The field of benchmarking products for group practices is still an immature market. However, several trends suggest such products are highly desirable. Provider organizations which bear medical risk need benchmarking data to help improve their efficiency. There are many important nonprovider organizations that need good information on group practices' utilization patterns and outcomes to help them plan new products and negotiate with physicians. PMID- 11067096 TI - TROT line: live and direct from the republic of Texas. PMID- 11067097 TI - The master clinician project. AB - This article describes an internal benchmarking process developed and used by the Marshfield Clinic targeting the interface between productivity and service quality. The benchmarking first identified "better performing" physicians using production and service quality measures as benchmarks. This was followed by detailed interviews of "better performers" to discover their "best practices." Based on an analysis of the "best practices" information, a physician curriculum was designed and implemented to improve service quality and provider productivity. Optimal strategies for successful programs are discussed and, finally, recommendations for future research are identified. PMID- 11067098 TI - Beware of patients under the influence. PMID- 11067099 TI - 20 respiratory challenges. Use training, technology & common sense to conquer respiratory problems. PMID- 11067100 TI - Heart failure and pulmonary edema. Understanding & correcting problems with the body's amazing pump. PMID- 11067101 TI - Tube test. Use the syringe aspiration technique to recognize esophageal intubations. PMID- 11067102 TI - An old drug with new tricks. Amiodarone hits the streets. PMID- 11067103 TI - Amiodarone: the cost of doing business. PMID- 11067104 TI - The pressure is on. Hyperbaric oxygen treatment as an adjunct to prehospital care. PMID- 11067105 TI - What to do about merger mania. PMID- 11067106 TI - Automated phone reminder systems can improve service and productivity. PMID- 11067107 TI - The economics of central billing offices. AB - The anticipation of economies of scale in physician billing has led many medical practices to consolidate their billing operations. This article analyzes these economies of scale, comparing performance indicators from centralized and decentralized operations. While consolidation provides compliance, control and information, diseconomies of scale can exist in the centralized receivables management process. The authors conclude that physician practices should consider a hybrid approach to billing, thus reaping the benefits of both centralization and decentralization. PMID- 11067108 TI - Seven steps to a profitable faculty practice plan. A case study in success at Connecticut Children's Medical Center. AB - A newly formed pediatric faculty practice plan had to overcome numerous hurdles blocking profitability, patient satisfaction, and time for clinical care, teaching and research. The authors describe the seven-point strategy that took the Connecticut Children's Medical Center Faculty Practice Plan from instability and disorganization to success as a high-volume clinical enterprise. PMID- 11067109 TI - Painless referrals. AB - Referrals, far and away the most burdensome administrative problem created by managed care, can be automated using secure, private intranets. Hundreds of primary care physicians and their offices in the Northwest have begun using this approach to reduce the approval time to get patients to specialists and to cut administrative costs. Combined with intranet delivery of laboratory and imaging test results, the service allows primary care physicians to fulfill their responsibility as overseers of the health care delivery process. PMID- 11067110 TI - Organized practice 2000 (and beyond): chaos or opportunity? PMID- 11067111 TI - The art and science of medical group turnarounds. Acting in several directions at once, acting at several points in time. AB - Developing a strategy and turnaround plan for a struggling medical group practice requires a combination of art and science. Orchestrating an effective about-face calls for decisive action on several fronts at once. Too many groups are slow to recognize the need to rescue a perilous situation. And rather than taking immediate action on multiple fronts, they try taking incremental steps over time. Implementing a successful turnaround requires both the diligence of follow through and the vision to make necessary adjustments throughout the process. PMID- 11067113 TI - "Back to the future". PMID- 11067112 TI - Creating a physician compact that drives group success. AB - A physician compact is the unwritten understanding of what an organization and its employees owe one another. When expectations clash about job security, salary, hours and the like, the practice--and its patients--suffer. Outdated expectations of the physician compact can hamper an organization from functioning smoothly and delivering optimal care. Rewriting a compact should involve all of a group's physicians, and should be endorsed and promoted by practice leadership. A satisfactory compact develops a shared sense of strategic imperatives and helps a practice reach its potential. PMID- 11067114 TI - AMA loses lead position on data safety. PMID- 11067115 TI - ASPs (application service providers) can be cure for cash crisis. PMID- 11067116 TI - Bridging the technical-clinical gap. Interview by John Morrissey. PMID- 11067117 TI - Using IT to make HMOs work. Interview by John Morrissey. PMID- 11067118 TI - Financial pro branches out. Interview by Andrew Pasternack. PMID- 11067119 TI - A pharmacist turns to IT. Interview by Barbara Kirchheimer. PMID- 11067120 TI - The career lessons of a CIO. Interview by Andrew Pasternack. PMID- 11067121 TI - Limits can advance e-health. PMID- 11067122 TI - Retooled venture resolves claims. PMID- 11067123 TI - The evolution of a CHIN (community health information network). PMID- 11067124 TI - Doctors tap into vital data. PMID- 11067125 TI - Transformation in small doses. PMID- 11067126 TI - Wanted: a Web geared to docs. PMID- 11067127 TI - Wash. hospital stays open, despite flaws. PMID- 11067128 TI - First in line. PMID- 11067129 TI - Flat earnings OK. PMID- 11067130 TI - Price-fixing or just good business? PMID- 11067131 TI - Docs seek antitrust relief from states. PMID- 11067132 TI - Ohioans could vote on HMO liability. PMID- 11067133 TI - Hospital food that draws 'em in. PMID- 11067134 TI - Food at Chicago-area hospital comes clean. PMID- 11067135 TI - HIMSS to create member subgroups. PMID- 11067136 TI - Survey: hospitals dabble in Internet. PMID- 11067137 TI - Calif. legislators seek seismic relief. PMID- 11067138 TI - Is MedPAC stacked? PMID- 11067139 TI - Integrity is the answer. PMID- 11067140 TI - Rebuilding from the rubble. AB - Much of the nursing home industry lies in financial ruins today, but that hasn't stopped a small but growing number of start-ups from moving in. Former executives of some of the for-profit nursing home companies that have filed for bankruptcy protection are hoping to pick up the pieces. They're using their industry expertise to launch companies backed by venture capital. PMID- 11067141 TI - Closing a loophole. PMID- 11067142 TI - Rising to the challenge. Interview by Scott Hensley. PMID- 11067143 TI - It's the little things that mean a lot. PMID- 11067144 TI - No kids abducted from hospitals in '99. . PMID- 11067145 TI - Bonds rebounding after a slow start. PMID- 11067146 TI - First transfer settlement. PMID- 11067147 TI - Discharge policy cuts hospital profits. PMID- 11067148 TI - The alternative model. AB - Consumers continue to believe in the power of unconventional medical treatments. They are expected to spend $31 billion on alternative and complementary therapies this year alone. But healthcare providers shouldn't see alternative medicine as an instant cash cow. The experts say it's best to start out small, as part of a seamless healthcare organization with established physician alliances. PMID- 11067149 TI - Trying, trying again. PMID- 11067150 TI - Leading indicators. PMID- 11067151 TI - HCFA rejects patient-restraints standard. PMID- 11067152 TI - An issue of 'quality'. PMID- 11067153 TI - Empire wins state OK. PMID- 11067154 TI - Battling medical errors Star Wars style. PMID- 11067155 TI - Caught in the fray. PMID- 11067156 TI - System divorces on rise. Unscrambling deals is messy and contentious. AB - The mass merger movement of the 1990s has hit the wall. More and more systems are taking a hard look at what their expensive deals have wrought, and are finding they don't like what they see. Disbanding systems are facing messy problems, and there are many questions as to whether de-merged hospitals can make it on their own. PMID- 11067157 TI - Making room for the laity. PMID- 11067158 TI - Rolling the dice in the Big Apple. PMID- 11067159 TI - More groups join plea for PPS delay. PMID- 11067160 TI - Top cardiovascular hospitals. PMID- 11067161 TI - What $75 million will get you. PMID- 11067162 TI - IRS sending message on novel bond deals. PMID- 11067163 TI - Ruling delivers good, bad news. PMID- 11067164 TI - Seeking better ways to improve patient safety. PMID- 11067165 TI - APC payment for hospitals to be effective on July 1. PMID- 11067166 TI - Well-planned policy for add-ons. PMID- 11067167 TI - Efficient method to schedule surgery. PMID- 11067168 TI - What it takes to manage an ASC. PMID- 11067169 TI - PACU staffing aided by point system. PMID- 11067170 TI - New Jersey hospital wins the battle, but loses the war. AB - Memorial Medical Center of South Amboy, N.J., was forced to close its doors and give up its license to operate as an acute care hospital. But a consortium of physicians and business people in the north central New Jersey city decided they couldn't do without the hospital's emergency service and attempted to reopen it. At the same time, the state's health department completed a report that found the state's 83 acute bed hospitals were "over-bedded" and refused to renew Memorial's license. This led to court actions and negotiations between the hospital and the state. The hospital's supporters mounted a major public relations blitz that reached the state capitol in Trenton. The campaign was successful in forcing negotiations, but the hospital was only allowed to provide emergency service as a satellite of a neighboring hospital. PMID- 11067171 TI - Options for Web sites changing fast in volatile health care market. PMID- 11067172 TI - Can the Internet solve all your hospital's recruiting problems? PMID- 11067173 TI - Solution brings greater patient satisfaction, higher visibility. PMID- 11067174 TI - Body Smart effort boosts community's health, centers' image. PMID- 11067176 TI - Sweet charity could be a pretty bitter pill. Will voters swallow prescription drug plans? PMID- 11067175 TI - Coordinated marketing, PR result in high profile for Arizona health center. PMID- 11067177 TI - From mumbo jumbo to a child's death. An unusual therapy and the risks it can pose. PMID- 11067178 TI - Building a better butt. Is it possible to make smoking safer? Or does that just tempt more smokers? PMID- 11067179 TI - Project Towards No Tobacco Use: implementation, process and post-test knowledge evaluation. AB - This paper describes the curricula contents, and presents data to evaluate the implementation, process and immediate post-test knowledge of Project Towards No Tobacco Use (Project TNT). Four different school-based tobacco use prevention curricula were developed to counteract the effects of three types of tobacco use acquisition variables typically addressed within a comprehensive social influences program: (1) peer approval for using tobacco (normative social influence), (2) incorrect social informational provided about tobacco use (information social influence) and (3) lack of knowledge or misperceptions about physical consequences resulting from tobacco use. Three curricula were designed to counteract the effects of single acquisition variables, whereas a fourth curriculum was designed to counteract the effects of combined social and physical consequences-related influences. These curricula were delivered to seventh grade students by trained project health educators to maximize implementation. 'Program' schools, those schools that received one of these curricula, were compared to 'control' schools that provided asystematic health education delivered by school personnel. A total of five conditions were contrasted through use of a randomized experiment involving 48 southern California junior high schools. This paper documents high levels of implementation in all program conditions. Also, favorable process ratings were obtained across the four program conditions, using multiple measures and sources of ratings (students, health educators and classroom teachers who observed curricula delivery). Finally, knowledge item sets completed by the students demonstrated discriminant validity across all five conditions. Because the program conditions were discriminable, yet were quite similar in implementation and process ratings, planned future study of behavioral outcomes can be interpreted as relatively uncontaminated by delivery or credibility confounds. PMID- 11067181 TI - The role of outcome and efficacy expectations in an intervention designed to reduce infants' exposure to environmental tobacco smoke. AB - The purpose of this paper is to examine the role of a theoretical framework in an intervention program designed to reduce infants' exposure to environmental tobacco smoke (ETS). The content of a nurse-based intervention focused on two psychosocial constructs: expectations of outcomes which may result from behaviors associated with ETS exposure and expectations of self-efficacy associated with the mother's ability to engage in these behaviors. This study found both constructs predictive of change in, and maintenance of, ETS exposure control. In particular, mothers reporting both low outcome and low efficacy expectations tended to have infants with the highest levels of ETS exposure. We also found that our intervention was effective in changing outcome and efficacy expectations in the desired direction. These findings suggest that outcome and efficacy expectations are changeable, and, therefore, represent important targets in future programs aimed at controlling ETS exposure. PMID- 11067180 TI - Background, conceptualization and design of a community-wide research program on adolescent alcohol use: Project Northland. AB - Project Northland is a community-wide research program funded by the National Institute on Alcoholism and Alcohol Abuse, for a 5-year period (1990-95). The aim of the study is to prevent or delay onset of alcohol use among young adolescents, as well as to reduce use among those who are already drinkers. Twenty communities were recruited in northeastern Minnesota, an area referred to as the Northland, Arrowhead or Iron Range region, and then were randomly assigned to either Education or Delayed Program conditions. The 10 Education school districts have agreed to participate in 3 years of intervention programs in schools, with parents and in the community-at-large. One group of young adolescents, the Class of 1998 (sixth grade students in the 1991-92 school year), form the study cohort. Surveys (1991-94) of the Class of 1998, their parents, community leaders and alcohol merchants are the primary components of the program's evaluation. Many conceptual and methodological questions emerged during the development of the research protocols for Project Northland over the past 2 years. These questions are the impetus for this article. Specifically, the focus on young adolescents and alcohol use was selected, as contrasted with older adolescents or with multiple problem behaviors. The project was designed using a community-wide model that addresses both supply and demand issues, rather than limited to a school based model. Intervention strategies and evaluation methods were chosen that could address community-level as well as individual-level behavior change, which required the development and application of new technologies. The rationale for these decisions may be useful to others considering community-wide health promotion efforts. PMID- 11067182 TI - Safer sex maintenance and reduction of unsafe sex among homosexually active men: a new therapeutic approach. AB - To date, publicly funded HIV/AIDS prevention efforts for homosexually active men have largely been limited to two traditional public health strategies: mass media information campaigns and HIV testing/contact notification programs. Health educators using either of these strategies have addressed the spread of HIV as they would many other infectious diseases and have relied heavily upon fear tactics or moral arguments to 'sell' the concept of safer sex. Grass-roots gay community efforts to prevent HIV transmission have also largely relied upon these two strategies, as well as upon more informal individual and group counseling activities. In general, however, strategies used by the gay community have tried to present more positive approaches to AIDS prevention, including eroticizing safer sex practices. This article reviews the efficacy of traditional public health approaches as well as the educational models underlying them, and argues that a major shift in focus is needed. A comprehensive health care and sexuality education model for homosexually active men based on the 'PLISSIST' sex therapy model is described and advocated. This model is presented as a more useful one for identifying populations at risk, reducing unsafe sexual behavior and promoting safer sex maintenance. The model identifies five sub-populations among homosexually active men and recommends specialized interventions appropriate to each. Adequate funding for the services included in this model is also advocated. PMID- 11067183 TI - Women defining health: food, diet and body image. AB - This article explores the relationship between gender and food through analysing data obtained from a series of women-only discussion groups, the participants of which explored self definitions of the term 'health'. What is distinct about this data, collected in the North East of England, is the clear link respondents perceive between food, diet and health. However, due to a number of factors such a knowledge may not result in dietary and ultimately bodily change. The many implications of such findings for health education are outlined in the discussion section of the paper. PMID- 11067184 TI - A prevention program for disturbed eating and body dissatisfaction in adolescent girls: a 1 year follow-up. AB - This study evaluated an intervention program to reduce moderate and extreme weight loss behaviors, disordered eating and low body image. The Body Image and Eating Behavior Intervention Program consisted of five specialized classes addressing media images of women, determinants of body size, healthy and unhealthy weight control methods, and emotional eating. The program was conducted in year 9 in Schools 1 (n = 80) and 2 (n = 27), while students from the same year in School 3 (n = 29) were control subjects. Participants completed self-report questionnaires assessing eating behavior and body image attitudes prior to the program, 1 months after the program and at 12 months follow-up, while control subjects completed the questionnaires at the same time but did not receive the program. Data were analyzed using groups (Schools 1, 2 and 3) by testing occasion (pre, post and follow-up) analyses of variance with repeated measures on testing occasion. Measures of disordered eating and frequency of use of extreme weight loss behaviors were constant over testing occasion and showed no effect of the intervention program. Body dissatisfaction was consistently lower in School 1 but increased across all subjects over the year. The implications of these data for school-based intervention programs in this area are examined. PMID- 11067185 TI - Back to complacency: AIDS in the Australian press, March-September 1990. AB - This paper reports the results of a quantitative and qualitative content analysis of all articles about AIDS published in the Australian metropolitan press during the 7 month period of March-September 1990. During the study period, almost 2800 articles mentioning AIDS were published, representing a drop in number of articles published compared with earlier years. Those issues receiving most press attention included people living with AIDS, AIDS and the law, AIDS policy and politics, the general spread of HIV/AIDS, AIDS education campaigns, drugs and medical treatment, and the HIV/AIDS threat posed to prison officers and health practitioners. The analysis demonstrates that the reporting of AIDS has changed over the course of the epidemic: topics which in the past commanded enormous press attention, such as AIDS as a 'gay plague' and the threat posed by the disease to heterosexuals, are no longer considered as newsworthy. Implications for AIDS health promotion activities are discussed. PMID- 11067186 TI - Development of a health education program for parents of preschool children with asthma. AB - This paper presents the development and pilot testing of a self-management education program for parents of preschool children (0-4 years) with asthma, involving general practitioners, asthma nurses, community nurses and doctors of child health centers. The program intends to integrate education in the medical care provided to the child (and the parent). The program contains four manuals, one for each group of health care providers, and a booklet for parents. The manuals identify the educational tasks per discipline and regulate referral from one discipline to another. The booklet provides written information for parents. In the development of the program, representative from both the target population and the providers of the education were involved in needs assessment surveys. Findings of these surveys were integrated into the design of the program. Then, a pilot study was conducted to test the efficacy of the program during group sessions. Findings indicate that the variables measured (knowledge, attitude, self-efficacy and self-management behaviors) improved significantly from pre- to post-test. Finally, the program was revised for the next phase in which the program will be evaluated in primary health care with a controlled trial. PMID- 11067187 TI - How young Europeans sleep. AB - This study investigated sleeping habits, difficulties in being able to fall asleep and their connections to self-reported health conditions, as well as other selected health behaviours and use of leisure time, among 11-16 year old Europeans from 11 countries. The study was part of a larger, comparative, WHO coordinated project on the health and life-style of school children (Health Behaviour of School Age Children--A WHO Cross-National Survey, The HBSC Study). In most of the countries, research data were collected from samples representative of the whole country. Using a standardized survey questionnaire, the data were collected anonymously in schools. Altogether 40,202 students responded to the survey. Sleeping habits and an inability to fall asleep varied significantly between countries as well as between age groups but only slightly between the sexes. Finnish school children experienced the most difficulty in being able to fall asleep. After the Israeli youth, the Finnish school children had the shortest night's sleep. At least a fifth of Finnish and Norwegian school children also reported that they felt tired almost every morning, the corresponding figure being smaller in other countries. A frequent use of psychoactive substances (alcohol and tobacco), lack of physical activity, excessive watching of TV/videos together with numerous evenings spent outside the home were all connected with going to bed late as well as with frequent difficulty in not being able to fall asleep. The results of the study offer an important challenge to health promotion and health education. Much more attention must be paid to this essential and exciting health habit! PMID- 11067188 TI - A community education monitoring system: methods from the Stanford Five-City Project, the Minnesota Heart Health Program and the Pawtucket Heart Health Program. AB - Understanding the process of behavior change interventions is critical to achieving campaign effectiveness and successful program replication. The present article presents a community education monitoring system (CEMS) using data from the Stanford Five-City Project (FCP), the Minnesota Heart Health Program (MHHP) and the Pawtucket Heart Health Program (PHHP). CEMS records the number and type of intervention activities, outcome objectives, targets of change (individual, organizational or environmental), channel(s) of dissemination and proportion of programs funded by the community. These data illustrate (1) the application of theory for each project, (2) data-based program administration, (3) feedback for revising programs and (4) type of reach or 'dose' information obtained from intervention monitoring. Process evaluations such as CEMS provide critical links between field realities and evaluation outcomes. This type of evaluation develops standards for measuring program reach and allows comparisons with other programs. CEMS also illustrates how programs enact theory. Validation studies are critical to the continued successful use of CEMS. The first step, however, is to develop a uniform way of describing complex multichannel behavior change programs. CEMS in a refined form should prove invaluable to health promotion program planners whether in research or service settings. PMID- 11067189 TI - Smoking habits of Grampian school children and an evaluation of the Grampian Smoke Busters campaign. AB - To determine the effect of a Smoke Busters club in reducing the numbers of children recruited to smoking, levels of smoking in Grampian amongst 11-14 year olds were determined at the launch of the club and again 22 months later, using a self-completion questionnaire. Initial smoking levels were similar to Scottish Office of Population Censuses and Surveys (OPCS) 1986 figures taken 2 years prior to the first Grampian survey. OPCS findings indicated that in Scotland smoking levels had remained fairly stable among girls and had increased among boys between 1986 and 1990. At the second survey in Grampian, smoking levels among girls and boys were lower than in Scotland generally, but it is not possible to know whether this was a result of the campaign or not. There was evidence of a relationship between the smoking habits of children and parents, particularly their mothers. Life-style questions showed that smokers were more likely to take part in activities characteristic of older teenagers and to prefer music which was associated with alternative, rebellious attitudes. Smoke Busters clubs aim to promote a non-smoking life-style amongst young teenagers. After 22 months of a Smoke Busters club, it was evident that at least 32.9% of the target group had joined the club at some point and that it was very popular with its members. Those who joined Smoke Busters were more than twice as likely to remain non smokers as those who were non-members, but this might not be a function of cause and effect. A further survey is to be undertaken at 4 years after the launch of the club and may allow assessment of any longer-term effects of the club in Grampian. PMID- 11067190 TI - Chronic fatigue syndrome: sociodemographic subtypes in a community-based sample. AB - Most chronic fatigue syndrome (CFS) studies are based on information about patients from primary or tertiary care settings. These patients might not be typical of patients in the general population. This investigation involved examinations of individuals with CFS from a community-based study. A random sample of 18,675 in Chicago was interviewed by telephone. Individuals with chronic fatigue and at least four minor symptoms associated with CFS were given medical and psychiatric examinations. A group of physicians then diagnosed individuals with CFS, who were then subclassified based on three sociodemographic categories--gender, ethnicity, and work status. Sociodemographic subgroups were analyzed in terms of symptom severity, functional disability, coping, optimism, perceived stress, and psychiatric comorbidity. Women, minorities, and nonworking individuals with CFS reported greater levels of functional disability, symptom severity, and poorer psychosocial functioning than men, Caucasians, and working individuals, suggesting sociodemographic characteristics may be associated with poorer outcomes in urban, community-based samples of CFS individuals. PMID- 11067191 TI - Key elements for implementing comprehensive health care models for persons with HIV: a stakeholder analysis. AB - A semistructured interview was conducted with 69 stakeholders in three university based health care projects that were funded to provide an integrated continuum of care for persons living with HIV/AIDS. Data from the key informant interviews yielded composite indicators of familiarity with the service model, the importance of the elements in the service model, and the perceived quality of services provided by these innovative HIV service demonstration projects. Ratings of service quality were related to ratings of the respondent's knowledge of the service demonstration project, the importance of the various elements in the service continuum, and several indicators of stakeholder characteristics using the data modeling method of Exhaustive CHAID (Chi-squared Automatic Interaction Detector). The groups of stakeholders most likely to give the highest quality or success ratings for these projects are identified. The implications of these findings for developing collaborative and comprehensive service models for persons with HIV/AIDS are discussed. PMID- 11067192 TI - The effects of clinical decision making on nurse practitioners' clinical productivity. AB - The degree of clinical decision making and clinical productivity among nurse practitioners (NPs) is of great interest to policy makers and planners involved in providing appropriate outpatient primary care services. The authors performed a statewide mailed survey of all NPs practicing either full-time or part-time in Wisconsin (response rate of 72.1%) to address the following research questions: Do the demographic characteristics, practice attributes, and primary practice settings of NPs impact their level of clinical decision making (e.g., the autonomy to order laboratory and radiological tests or to refer a patient to a physician specialist other than their collaborating physician)? Do NPs' levels of clinical decision making correlate with their outpatient clinical productivity, adjusting for demographic characteristics, practice attributes, and primary practice settings? The multiple linear regression results indicated that having more years in practice as an NP, practicing in the family specialty area (vs. a combined other category, which included pediatrics, acute care, geriatrics, neonatal, and school), treating patients according to clinical guidelines, practicing in settings with a fewer number of physicians, and practicing in a multispecialty group practice versus a single-specialty group practice were associated with greater levels of clinical decision making. However, NPs who primarily practiced in a hospital/facility-based practice, as compared with a single-specialty group practice, had lower levels of clinical decision making. After adjusting for demographic characteristics, practice attributes, and primary practice settings, NPs with greater clinical decision-making authority had greater outpatient clinical productivity. The conclusions discuss the policy implications of the findings. PMID- 11067193 TI - Health-related physical fitness knowledge of student allied health professions. AB - Each allied-health profession has their own particular expertise but also shares some commonalities. One such commonality should be knowledge of health-related physical fitness relating to the health and well-being of individuals. Although the benefits of health-related physical fitness have been well documented, few studies have examined the level of health-related physical fitness knowledge among allied-health professions. Therefore, the purpose of this investigation was to assess the health-related physical fitness knowledge of three allied health professions using a 40 item multiple-choice test designed to assess knowledge in five domains of health-related physical fitness. Results indicated that student athletic trainers scored significantly higher on the post-test versus pre-test. On the post-test, athletic training and physical therapy groups scored significantly higher than the nursing group. The information from this study may be valuable in aiding educators in developing appropriate curricula to better prepare students for their role as allied health professionals. PMID- 11067194 TI - An eclectic model for evaluating web-based continuing medical education courseware systems. AB - World Wide Web and compact disc-read only memory technologies have introduced new prospects for delivering continuing medical education (CME) to rural and remote physicians. However, evidence concerning the effectiveness of these technologies in providing CME, and approaches to their evaluation, is limited. The rationale for this study was to design a model for evaluating the effectiveness of computer mediated CME courseware. An eclectic, evaluation-planning matrix was designed by selecting various concepts from the literature and was used in planning and developing the evaluation model. The model was field-tested by evaluating a computer-mediated courseware program on dermatological office procedures, and a meta-evaluation was conducted to assess the effectiveness of the evaluation methods and procedures. The findings suggest that the model was useful in collecting data to inform decision making and to improve the instructional product. The field test results revealed that computer-mediated instruction was effective in delivering CME at a distance. PMID- 11067195 TI - Encouraging physicians to respond to surveys through the use of fax technology. AB - High response rates in surveys of physicians are difficult to achieve. One possible strategy to improve physicians' survey participation is to offer the option of receiving and returning the survey by fax. This study describes the success of the option of fax communication in a survey of general practitioners, family physicians, and pediatricians in Arkansas with regard to pediatric asthma. Eligible physicians were given the choice of receiving the survey by telephone, mail, or fax. In this observational study, physicians' preferences, response rates, and biases for surveys administered by fax were compared with mail and telephone surveys. The overall survey response rate was 59%. For the 96 physicians completing an eligibility screener survey, the largest percentage requested to be surveyed by fax (47%) rather than by telephone (28%) or mail (25%). Faxing may be one strategy to add to the arsenal of tools to increase response rates in surveying physicians. PMID- 11067196 TI - Student attitudes toward same-gender versus mixed-gender partnering in practicing physical examination skills. AB - Students were surveyed to assess perceptions with regard to the use of same gender and mixed-gender partnering in practicing physical examinations. There was support for this arrangement, with students reporting that the practice provided an important learning experience with reasonable levels of comfort and no loss of thoroughness and rigor. Students did express some concerns. Females tended to have lower comfort levels, particularly when they were in the role of patient. Many of the concerns expressed could be addressed with procedural changes during the assignment of partners, and the benefits of continuing this practice seem to outweigh any negative aspects. PMID- 11067197 TI - Rising stars. PMID- 11067198 TI - Feeding frenzy. What kind of market will emerge when the consolidation wave settles? PMID- 11067199 TI - The many layers of workflow automation. PMID- 11067200 TI - Cyberspace: an uncertain domain for clinical labs. PMID- 11067201 TI - Banking on good healthcare. PMID- 11067202 TI - Claims processing speeds up. AB - Healthcare providers, payors and patients alike are somewhat dubious about using the Web to administer transactions. Everyone agrees that it would save time and money. But the real-time exchange of funds inherent to e-business would require payors and patients to settle up front. And technology can make employees uncomfortable, not to mention obsolete. PMID- 11067203 TI - Healthcare Informatics 100. PMID- 11067204 TI - Toward more perfect integration. Standard-bearers march on. PMID- 11067205 TI - Search for strategies to bring Australians together. PMID- 11067206 TI - Is wireless really there? Systems integrators discuss what CIOs should know before they journey into the realm of wireless. PMID- 11067207 TI - Consultants: finding the right fit. E-business initiatives add another piece to the IT puzzle. PMID- 11067208 TI - Healthcare IT consultants ... a sampling. PMID- 11067209 TI - One for all, all in one. PMID- 11067210 TI - The rudder in a white-water world. PMID- 11067211 TI - The return on investment crunch. PMID- 11067212 TI - Live birth and infant death record linkage: methodological and policy issues. AB - Using records from Ohio annual vital statistics tapes, we describe a method for linking live birth and infant death certificates and for dealing with late registered and unregistered births. In our 1985-87 Cleveland and East Cleveland study population, deceased infants with late-registered births were found to be similar to those with timely registered births. Approximately 4.6% of decedents, however, had unregistered births and these tended to be very premature infants from socially disadvantaged backgrounds who died shortly after delivery (including homicides following birth at home). We discuss the policy implications of failing to link infant deaths with unregistered births in studies of birth outcomes. PMID- 11067213 TI - Managed behavioral health care provider practice patterns: a new item for the public policy agenda. AB - Managed care is a dominating issue on the public policy agenda. Difficulties in defining and operationalizing it continue to have ramifications for the nation. It is often assumed that the care being reimbursed by managed care organizations is for clients whose psychiatric conditions have been appropriately diagnosed and treated. Based on the responses of a randomly-selected group from the major behavioral health care disciplines, not all care reimbursed is for care which has been appropriately diagnosed and treated. The cost implications of managed care and the ramifications for public health policy are discussed. PMID- 11067214 TI - Addressing barriers to community care of battered women in rural environments: creating a policy of social inclusion. AB - Given domestic violence continues to be a serious social problem, recently acknowledged as a national public health epidemic as well, the need to examine existing formal helping services and address the barriers to service utilization among battered women is warranted. Social exclusion is the process by which multi dimensionally disadvantaged individuals are prohibited from obtaining formal helping services. Although not a policy per se, many community helpers and organizations inadvertently contribute to erecting barriers which exclude battered women from using such services, particularly in rural environments. Suggestions to transform a process of social exclusion to social inclusion for battered women is offered. PMID- 11067215 TI - What patients know about their rights in Turkey. AB - Patients' rights issues are currently being discussed in health care at both the private and public levels in Turkey. In this study, 317 patients were interviewed upon hospital discharge and asked what they knew about their "rights" as patients. According to the collected data, 63% of the patients were not aware that they had any rights in receiving health care services at all. Since this is the first study of its type in Turkey, further research in this area is needed. In closing, it is recommended that hospitals in Turkey adapt routine policies similar to those in the United States for informing customers about their rights for safe, effective and efficient health care provision. PMID- 11067216 TI - Use of psychedelics among Miami's public school students, 1992. AB - This analysis examined psychedelic use by 475 adolescents in Dade County, Florida public schools in 1992. Significant factors which increase the probability of use include peers' use of psychedelics, the fact that adolescents are White, and their involvement in music-related activities at school. This descriptive account is based on probit analysis and attempts to isolate factors related to psychedelic use. PMID- 11067217 TI - Military hospitals join forces to combat inefficiency. PMID- 11067218 TI - Robodoc. Robotics: brave new world of surgery--or is it? PMID- 11067219 TI - Comeback kids. Cost control tool spurs hospital turnaround. PMID- 11067220 TI - Premier pilots aim to reap the benefits of UPN. Raising the bar. PMID- 11067221 TI - Heads up! Pay what you owe--and not a penny more. PMID- 11067222 TI - Workplace wars. How to take the bite out of the fight. PMID- 11067223 TI - Stats. How do bosses view you? PMID- 11067224 TI - Outsourcing. In or out? PMID- 11067225 TI - Rules & regs. Roundup '99. PMID- 11067226 TI - The Midas touch. Turn Y2K expenditures into MEMS gold. PMID- 11067228 TI - Stats. Price rise forecast: it's a rerun. PMID- 11067227 TI - Clean scene. New processes bring sweeping changes. PMID- 11067229 TI - The pressure's on. A hypertension drug taken by 28 million people is under scrutiny. What are the other options? PMID- 11067230 TI - Are drugs winning the games? PMID- 11067231 TI - Flight nurse research activities. AB - INTRODUCTION: The study purpose was to identify how members of the Air & Surface Transport Nurses Association (ASTNA), formerly called the National Flight Nurses Association, use research in their flight nursing practice. METHODS: A survey was mailed to each ASTNA member requesting demographic data and a description of research activities used in their practice of flight nursing. RESULTS: Thirty percent of the 1643 members (n = 497) completed the survey. Sixty-four percent (320) of respondents were direct patient care providers, and 47% were certified flight registered nurses (CFRN). Thirteen percent (65) reported that research was mandated in their job description. Sixty-three percent (315) used research to guide practice, and 37% (183) incorporated research into policies and procedures. Thirty percent (149) actively conducted some type of research, 15% (76) presented at national conferences, and 11% (53) published in professional journals. CONCLUSION: Components of the ASTNA research standard are being met by most flight nurses. However, only a small percentage of these nurses are designing and conducting studies and formally disseminating their research findings. Flight nurses are involved in research, but their overall participation could be improved. PMID- 11067232 TI - Storage temperatures of medications on an air medical helicopter. AB - INTRODUCTION: The safety and efficacy of medications stored on air medical helicopters may be adversely affected by extreme temperatures. The purpose of this study was to determine whether temperatures inside an air medical helicopter drug box were within the U.S. Pharmacopeia recommendations for controlled room temperature. This is defined as a temperature between 15 degrees and 30 degrees C (59 degrees and 86 degrees F) with a mean kinetic temperature of less than 25 degrees C (77 degrees F). An additional goal was to determine whether time/temperature indicator labels can reliably monitor mean kinetic temperatures. METHODS: Temperatures were monitored with miniature electronic temperature recorders and color-changing time/temperature indicator labels. RESULTS: The mean kinetic temperatures for the summer and winter periods were 25.1 degrees C (77.2 degrees F) and 12.7 degrees C (54.8 degrees F), respectively. In the summer, the electronic recorders logged temperatures exceeding 25 degrees C (59 degrees F) 37% of the time and more than 30 degrees C (86 degrees F) 6% of the time. In the winter, temperatures less than 15 degrees C (59 degrees F) were recorded 83% of the time. The mean kinetic temperatures obtained from the electronic recorder and the time/temperature indicator labels differed by less than 0.7 degree C (1.3 degrees F). The results show that medications on an air medical helicopter are subject to temperatures out of the recommended range and that time/temperature indicator labels can reliably monitor mean kinetic temperatures. PMID- 11067233 TI - Use of local climatic data to determine if weather precludes the operation of an air medical system. AB - INTRODUCTION: Weather is one of many factors that affect safety in an air medical program. Syracuse, New York, has notoriously bad weather, and some have questioned whether an air medical service is practical given central New York's climate. This study was undertaken to determine the extent to which the area's climate could be expected to limit the availability of an air medical service. METHODS: CAMTS weather minimums for rotor-wing programs were compared with 1996 1997 hourly weather observations from the Northeastern Regional Climate Center (NRCC) and sunrise/sunset data from the United States Naval Observatory to determine how frequently weather conditions could be expected to preclude an air medical response in the greater Syracuse area. RESULTS: Exactly 17,544 hourly observations were made. CAMTS weather minimums would have precluded local flights for 606 (3.5%) of these hours and cross-country flights for 1111 (6.3%) hours. Cross-country flights were more likely to be precluded than local flights (P = .001), and both local and cross-country flights were more likely to be precluded at nighttime than in the daytime (P = .001). All flights were more likely to be precluded during winter months than during summer months (P = .000). CONCLUSION: The weather in central New York generally does not preclude the operation of an air medical services system. PMID- 11067234 TI - Atlantooccipital disassociation: a case report. PMID- 11067235 TI - Cause factor: human. PMID- 11067236 TI - Incubator heat loss. PMID- 11067237 TI - Rotor-wing transport of patients with a biventricular assist device. PMID- 11067238 TI - A national survey of air medical infectious disease control practices. AB - INTRODUCTION: Caring for an infectious patient in the air medical environment presents a special challenge to all air crew members (ACMs) involved. The purpose of this study was to survey the infectious disease control practices of air medical programs (AMPs) that are members of the Association of Air Medical Services. METHODS: A structured telephone survey was designed to gather data. Using one interviewer (an undergraduate student) with no knowledge of the study's goal minimized experimental bias. AMPs from 151 geographically selected areas were called between June and August 1996. Only the programs' chief flight nurses (CFNs) were targeted as respondents. RESULTS: The response rate was 91% (138 of 151). Although no program refused to participate, 13 CFNs were unavailable to be interviewed. Mission profile was 32% scene and 68% interhospital with an annual average of 950 patient transports per program. Transport type was 61% rotor-wing aircraft, 17% fixed-wing, and 22% both. Flight physicals for ACMs were required by 57% of the AMPs. Pre-employment screenings for rubella, tuberculosis (TB), and varicella were noted. Interestingly, 17% of the AMPs reported pre-employment HIV testing. Immunization was mandated by 57% of AMPs, including hepatitis B virus, measles, rubella, and tetanus. Nine percent of the respondents refused to accept a transport with specific contagious conditions, primarily TB. A formal decontamination policy was in effect at 88% of the AMPs, and OSHA-approved filter masks were available at 70%. Pathogen exposure reporting was required by 97%. CONCLUSION: A current, comprehensive infection control program, continuing education, and 100% compliance with standard precautions will help reduce the possibility of accidental exposures. These strategies to reduce transmission also can be extended during training sessions to the prehospital and hospital personnel with whom the air medical program serves. PMID- 11067239 TI - Have a cookie: how your website should respond to the DoubleClick controversy. PMID- 11067240 TI - Impedance cardiography (ICG) noninvasive hemodynamic monitoring provides an opportunity to deliver cost effective, quality care for patients with cardiovascular disorders. AB - Hemodynamic evaluation is an essential component in diagnosing cardiovascular disorders and managing patient care. There are numerous invasive, semi-invasive, and noninvasive methods available for evaluating hemodynamic function, which vary widely in equipment, procedural complexity, cost, quality of data, and risks. This paper summarizes information needed for administrative decision-making about employing one method--ICG hemodynamic monitoring--and provides an overview of the technology, its accuracy and reproducibility, clinical applications, financial consideration and references for further evaluation of the technology. PMID- 11067241 TI - Will insurers help pay for end-of-life care? PMID- 11067242 TI - Depression in the workforce. Part 2: The best defense. PMID- 11067243 TI - Are drugs a cost or an investment? PMID- 11067244 TI - Health insurers play hardball with brokers. PMID- 11067245 TI - Helping employees battle the bulge. PMID- 11067246 TI - Why work where you can't get coverage? PMID- 11067247 TI - DataWatch. Health insurance fraud busters. PMID- 11067248 TI - Do patients matter? Contribution of patient and care provider characteristics to the adherence of general practitioners and midwives to the Dutch national guidelines on imminent miscarriage. AB - OBJECTIVE: To assess the relative contribution of patient and care provider characteristics to the adherence of general practitioners (GPs) and midwives to two specific recommendations in the Dutch national guidelines on imminent miscarriage. The study focused on performing physical examinations at the first contact and making a follow up appointment after 10 days because these are essential recommendations and there was much variation in adherence between different groups of providers. DESIGN: Prospective recording by GPs and midwives of care provided for patients with symptoms of imminent miscarriage. SETTING: General practices and midwifery practices in the Netherlands. SUBJECTS: 73 GPs and 38 midwives who agreed to adhere to the guidelines; 391 patients were recorded during a period of 12 months. MAIN MEASURES: Adherence to physical examinations and making a follow up appointment were measured as part of a larger prospective recording study on adherence to the guidelines on imminent miscarriage. Patient and care provider characteristics were obtained from case recordings and interviews, respectively. Multilevel analysis was performed to assess the contribution of several care provider and patient characteristics to adherence to two selected recommendations: the number of recommended physical examinations at the first contact and the number of days before a follow up appointment took place. RESULTS: In the multilevel model explaining variance in adherence to physical examinations, the care provider's acceptance of the recommendations was the most important factor. Severity of symptoms and referral to an obstetrician were significant factors at the patient level. In the model for follow up appointments the characteristics of the care provider were less important. Referral to an obstetrician and probability diagnosis were significant factors at the patient level. CONCLUSIONS: The study showed that characteristics of both the patient and care provider contribute to the variability in adherence. Furthermore, the contribution of the characteristics differed per recommendation. It is therefore advised that the contribution of both patient and care provider characteristics per recommendation should be carefully examined. If implementation is to be successful, strategies should be developed to address these specific contributions. PMID- 11067249 TI - Organisational culture and quality of health care. PMID- 11067250 TI - Organisational sources of safety and danger: sociological contributions to the study of adverse events. AB - Organisational sociology has long accepted that mistakes of all kinds are a common, even normal, part of work. Medical work may be particularly prone to error because of its complexity and technological sophistication. The results can be tragic for individuals and families. This paper describes four intrinsic characteristics of organisations that are relevant to the level of risk and danger in healthcare settings--namely, the division of labour and "structural secrecy" in complex organisations; the homophile principle and social structural barriers to communication; diffusion of responsibility and the "problem of many hands"; and environmental or other pressures leading to goal displacement when organisations take their "eyes off the ball". The paper argues that each of these four intrinsic characteristics invokes specific mechanisms that increase danger in healthcare organisations but also offer the possibility of devising strategies and behaviours to increase patient safety. Stated as hypotheses, these ideas could be tested empirically, thus adding to the evidence on which the avoidance of adverse events in healthcare settings is based and contributing to the development of theory in this important area. PMID- 11067251 TI - Developing valid cost effectiveness guidelines: a methodological report from the north of England evidence based guideline development project. PMID- 11067253 TI - Improving care for people with diabetes. PMID- 11067252 TI - Complications of diabetes: screening for retinopathy and management of foot ulcers. PMID- 11067254 TI - Managing quality in primary health care: the need for valid information about performance. PMID- 11067255 TI - Are routine outcome measures feasible in mental health? PMID- 11067256 TI - Inequalities in access to diabetes care: evidence from a historical cohort study. AB - OBJECTIVE: To establish which factors predict attendance at a hospital diabetes clinic and for diabetes review in general practice. DESIGN: A historical cohort study of individuals with diabetes identified from general practice records. Information on service contacts and other clinical, social, and demographic variables was collected from general practice records and postal questionnaires. SETTING: Seven Leicestershire general practices. SUBJECTS: Individuals registered with study practices who had a diagnosis of diabetes made before 1990. MAIN OUTCOME MEASUREMENTS: Attendance at a hospital diabetes clinic or for a documented diabetes review in general practice at least once between 1990 and 1995. RESULTS: 124 (20%) had at least one recorded diabetes review in general practice and 332 (54%) attended a hospital diabetes clinic at least once. The main predictors of attending a hospital clinic were younger age, longer duration of diabetes, and treatment with insulin. Access to a car (OR 1.34, 95% CI 1.06 to 1.71), home ownership (OR 1.48, 95% CI 1.14 to 1.58) and a non-manual occupation (OR 1.56, 95% CI 1.09 to 2.24) were all associated with an increased likelihood of attending, although living in a less deprived area was not. The main predictors of attending for review in general practice were older age, less co morbidity, and being white. Living in a more deprived area was related to a reduced chance of review in general practice (OR 0.81, 95% CI 0.76 to 0.86) while individual socioeconomic indicators were not. CONCLUSIONS: Whilst an indicator of area deprivation predicts reduced likelihood of review in general practice, individual indicators predict reduced likelihood of attending outpatients. This suggests a need for different approaches to tackling inequalities in access to care in primary and secondary care settings. PMID- 11067257 TI - Clinical governance in primary care groups: the feasibility of deriving evidence based performance indicators. AB - OBJECTIVES: To test the feasibility of deriving comparative indicators in all the practices within a primary care group. DESIGN: A retrospective audit using practice computer systems and random note review. SETTING: A primary care group in southern England. SUBJECTS: All 18 general practices in a primary care group. MAIN OUTCOME MEASURES: Twenty six evidence-based process indicators including aspirin therapy in high risk patients, detection and control of hypertension, smoking cessation advice, treatment of heart failure, raised cholesterol levels in those with established cardiovascular disease, and the treatment of atrial fibrillation. Feasibility was tested by examining whether it was possible to derive these indicators in all the practices; the problems and constraints incurred when collecting data; the variations in indicator values between practices in both their identification of diseases and in the uptake of various interventions; the possible reasons for these variations; and the cost of generating such indicators. RESULTS: It was possible to derive eight indicators in all practices and in three practices all 26 indicators. The median number of indicators derived was 12 with two practices able to generate eight. There was considerable variation in the use of computers between practices and in the ability and ease of various practice computer systems to generate indicators. Practices varied greatly in the identification of diseases and in the uptake of effective interventions. Variation in identification of ischaemic heart disease could not be explained by a higher prevalence in practices with a more deprived population. The cost of generating these indicators was 5300 Pounds. CONCLUSION: Comparative evidence-based indicators, used as part of clinical governance in primary care groups, could have the potential to turn evidence into everyday practice, to improve the quality of patient care, and to have an impact on the population's health. However, to derive such indicators and to be able to make meaningful comparisons primary care groups need greater conformity and compatibility of computer systems, improved computer skills for practice staff, and appropriate funding. PMID- 11067258 TI - Validation of an outcome scale for use in adult psychiatric practice. AB - OBJECTIVE: To clarify the usefulness, acceptability, sensitivity, and validity of version 4 of the Health of the Nation Outcome Scale (HoNOS), a scale developed to meet the requirement for a clinically acceptable outcome scale for routine use in mental illness services. DESIGN: Patients with a range of mental illnesses were rated on the HoNOS at the beginning and end of an episode by interviews with mental health professionals. SUBJECTS: 934 patients from eight diagnostic categories were rated by 129 mental health professionals at 17 sites; 250 were also rated on a range of comparison scales. OUTCOME MEASURES: Comparison of patients' scores at the beginning and end of an episode using individual item scores, dimensional subscores, and the total score. RESULTS: HoNOS scores decreased by almost 50% between the beginning and end of episodes. They varied with the severity of the setting and discriminant analysis showed that the HoNOS had a moderate level of discriminatory power. Correlation analysis showed acceptable levels of agreement with independent scales, although the accuracy of ratings of some items at the beginning of an episode was affected by information deficits. CONCLUSION: The findings indicate that HoNOS is sensitive to change across time and to differences in illness type and severity, and has a sufficient degree of both construct and criterion related validity to fulfil the requirements of a mental health outcome scale for routine use in clinical settings. PMID- 11067259 TI - The drug charade. PMID- 11067260 TI - Grace under pressure ... gyrotonics. PMID- 11067261 TI - Focus on evaluating CPR systems. Critical success factors. PMID- 11067262 TI - The need for dedicated system coordinators. PMID- 11067263 TI - Clicking the way to success. PMID- 11067264 TI - Checklist to automate your claims process. PMID- 11067265 TI - PDAs and hand-helds: world without wires. PMID- 11067266 TI - ASPs: healthcare reaches back to the future. PMID- 11067267 TI - Staffing and the quality of care. PMID- 11067268 TI - 100 top hospitals. Conversations with four captains of industry. PMID- 11067269 TI - Integrated systems vs. plug-and-play. PMID- 11067270 TI - Web interface is a two-way street. PMID- 11067271 TI - Streamlining the flow of information. PMID- 11067272 TI - HotList. CPR/EMR. PMID- 11067273 TI - Involving physicians in developing IT solutions. PMID- 11067274 TI - This isn't kids' stuff. PMID- 11067275 TI - Building room to grow. PMID- 11067276 TI - Enhancing quality through integrating the enterprise. PMID- 11067277 TI - Collaboration in the name of national standards. PMID- 11067278 TI - Seclusion and restraint: leaders are still polarized. Dialogue. PMID- 11067279 TI - Will we ever be on the same page? PMID- 11067281 TI - Sounding a call to action. PMID- 11067280 TI - Our quality-assurance methods aren't so sure. PMID- 11067282 TI - Try these quality improvement worksheets. PMID- 11067283 TI - The future looks just as complicated. PMID- 11067284 TI - CMHCs stretch the limits. PMID- 11067285 TI - Six trends to shape the field's future. PMID- 11067286 TI - It's not easy directing this production. PMID- 11067287 TI - Will ORYX take you to the top? PMID- 11067288 TI - Leadership: do you have what it takes? PMID- 11067289 TI - Outcomes behind bars. Corrections systems overcome challenges to meaningful evaluation. PMID- 11067290 TI - Are all SSRTIs created equal? PMID- 11067292 TI - ADHD: improving diagnosis in primary care. PMID- 11067291 TI - Drugs produce results ... and there's more to come. PMID- 11067293 TI - A Texas-sized prescription. PMID- 11067294 TI - Fiddlin' around in Arkansas. PMID- 11067295 TI - 2000 Resource Guide. Associations. PMID- 11067296 TI - 2000 Resource Guide. Government organizations. PMID- 11067297 TI - 2000 Resource Guide. Standards groups. PMID- 11067298 TI - Health care I.T.: 2000 and beyond. PMID- 11067299 TI - In the pipeline. PMID- 11067300 TI - Assimilation complication. PMID- 11067301 TI - For health care CIOs, a brave new world. PMID- 11067302 TI - Acquisitions can be tough to swallow. PMID- 11067303 TI - For vendors, HIPAA offers opportunities. PMID- 11067304 TI - Quality data for quality care. PMID- 11067305 TI - Dr. Holly goes to Hollywood. PMID- 11067306 TI - Getting everyone involved. Interview by Beckie Kelly. PMID- 11067307 TI - CIO secrets to calculating return on investment. PMID- 11067308 TI - Deploying an I.T. cure for chronic diseases. PMID- 11067309 TI - CIO's best strategies to prepare for disaster. PMID- 11067310 TI - The best laid plans of HMOs. PMID- 11067311 TI - Time studies identify interventions to boost patient flow in your ED. AB - According to researchers who completed a five-year study at the Olive View University of California at Los Angeles Medical Center in Sylmar, CA, time studies are the most effective way to identify delays and improve efficiency in the ED. Time studies may reveal that your assumptions about delays are incorrect. A five-year study showed a connection between the median waiting time to see a physician and the number of patients who leave without being seen. Constructing a time study allows you to understand all the steps in patient care. Identify who is responsible for each increment of time, including the registration clerk, bed control staff person, nurses, X-ray personnel, lab staff, and physicians. PMID- 11067312 TI - Special report: resolving conflicts. Resolve conflicts between ED and other departments. AB - Physicians and nurses have a tendency to avoid conflict, but ED staff must learn strategies to manage common conflicts that arise with other departments. There are often conflicts over whether patients should be admitted or which service admitted patients should be sent to. On-call physicians often focus on peripheral issues in patient care, which can seem patronizing to ED staff. If ED nursing leadership reports to the same director as critical care nurses, both departments will have shared goals with less chance of conflict. Educate staff in other departments that 30% to 40% of hospital admissions come through the ED. PMID- 11067313 TI - Special report: resolving conflicts. What to do about tension with other departments. PMID- 11067314 TI - It's finally here. Special bulletin clarifies EMTALA regulations. AB - The Office of the Inspector General and the Health Care Financing Administration have published a special advisory bulletin warning that you must not delay medical screening exams or stabilization, even when managed care plans require prior authorization. Medicare and Medicaid managed care organizations were put on notice that the practice of requesting prior authorization is illegal under the Emergency Medical Treatment and Labor Act (EMTALA) and the Balanced Budget Act. You can ask about payment after stabilization has commenced. Only staff knowledgeable about EMTALA should answer financial questions from patients. Consistently long waits could be interpreted as a pattern of encouraging patients to leave without treatment. PMID- 11067315 TI - Get ready for new pain management standards. PMID- 11067316 TI - Beware: dual staffing is legal but risky. PMID- 11067317 TI - Know 6 key points of EMTALA bulletin. PMID- 11067318 TI - ACEP's 2000 goal: access for the uninsured. AB - The growing problem of the uninsured population may be the next major issue the American College for Emergency Physicians (ACEP) will focus on in 2000, after prudent layperson legislation is passed and enforced in all states. Prudent layperson definition standards have been adopted in 28 states, but ACEP is now focusing on making sure those laws are adequately enforced. ACEP's Emergency Practice Committee is evaluating telephone advice and patient satisfaction survey tools. ACEP is developing an extended course to teach physicians management skills. PMID- 11067319 TI - Update on patient access to EDs: share good news. AB - The Norwood/Dingell bill, passed in the House of Representatives, would establish a national standard of emergency care and protect patients from claims denials and managed care plans' requirements for prior authorization. The bill will be a significant health care issue in this year's presidential election, but it's doubtful that federal legislation will be passed before then. It would establish a national prudent layperson standard, which requires health plans to base emergency care coverage on symptoms and not a final diagnosis. Patients who pay for their health insurance need the same emergency care protections as individuals enrolled in federal government, Medicare, and Medicaid health plans. PMID- 11067320 TI - Expect 2 waves of patients after terrorist attack. AB - A study examining patient arrival at local EDs after the bombing of the federal building in Oklahoma has shown there are two "waves" of patients after a terrorist attack. Contrary to popular belief, after a mass casualty incident, most patients arrive by means other than ambulance transport. Because the first group of patients arriving at EDs are transporting themselves, they tend to be of less severity than the second wave of patients. While triaging patients from the first wave, you need to prepare for more serious injuries in the second. PMID- 11067321 TI - Outpatient PPS switch likely to be delayed. AB - Congress approved a Medicare bill that likely will delay implementation of the final rule from the Health Care Financing Administration on ambulatory patient classifications. Even a one-year time period may not be enough to improve documentation adequately, so ED managers must start preparing now. Consider investing in documentation improvement tools, such as template paper records, disk transcriptions, and computerized emergency medical records. PMID- 11067322 TI - Should you vaccinate patients in your ED? Check out these unique protocols. AB - Although controversial, ED vaccination programs can successfully target minority, low-income, and uninsured populations. Without access to vaccination registries, only single-shot immunizations such as influenza and pneumococcal are feasible in the ED. For a vaccine program to be financially feasible, an ED needs a significant Medicaid population and low immunization rates in the community. Protocols for vaccine programs should delegate the responsibility to nursing with standing orders. PMID- 11067323 TI - ED nurses may resist vaccination programs. PMID- 11067324 TI - ED managers: teamwork is key to success with APCs. AB - Collaboration with other departments is key to preparing for the switch to ambulatory payment classifications (APCs) proposed by the Health Care Financing Administration. Physician documentation will be critical to the overall success of your hospital's experience with APCs. Late charges caused by hospital-related billing information depending on the physician diagnosis will not be paid. The APC system will require better cooperation among all hospital departments, including medical records and the billing office. PMID- 11067325 TI - Know difference between APCs, current system. PMID- 11067326 TI - Joint Commission IDs five high-alert meds. AB - The Joint Commission on Accreditation of Healthcare Organizations has issued a bulletin listing "high-alert" medications that have the highest risk of causing injury when misused. The five high-alert medications are insulin, opiates and narcotics, injectable potassium chloride (or phosphate) concentrate, intravenous anticoagulants (heparin), and sodium chloride solutions above 0.9%. The Joint Commission recommends strategies such as a system that confirms the correct drug, dosage, patient, time, and route. PMID- 11067327 TI - Synchronize timepieces in your trauma room. AB - Timepieces are probably inaccurate in your ED, and that can have serious consequences during critical events such as major trauma and cardiac arrest resuscitation. During resuscitations, multiple timepieces are used, and many events occur in a short period of time. Inaccuracies make it impossible to reconstruct the order of events accurately, which increases liability risks in the event of a lawsuit. Staff can synchronize their timepieces by calling the local number that announces the atomic clock time every 10 seconds; ideally, however, all clocks should be set universally by a central server. PMID- 11067328 TI - Emergency etiquette. Minding your safety Ps and Qs. PMID- 11067329 TI - Strokes. Misconceptions, prevention, warning signs ... and the right response. PMID- 11067330 TI - Ready to answer ORYX data questions? Here's what Joint Commission will ask. AB - The Joint Commission on Accreditation of Healthcare Organizations' ORYX program integrates performance measurement into the accreditation process. Your hospital already should have chosen six measures to report for the ORYX program, and those measures may affect the ED. Surveyors will want to see comparison charts, so your ED can be measured against other facilities, and control charts with internal data. Automatic type I recommendations result if institutions fail to report data they said they would report for two quarters in a row. Your control charts may have findings that are statistically, but not clinically, significant. PMID- 11067331 TI - A union for ED physicians is not the solution. PMID- 11067332 TI - Labor organization is a boon for physicians. PMID- 11067333 TI - Here are 5 ways to celebrate EMS Week. PMID- 11067334 TI - HCFA's final rule on APCs is out: ED managers can breathe sighs of relief. AB - The final rule on ambulatory payment classifications (APCs) for outpatient services from the Health Care Financing Administration (HCFA) was published in the April 7, 2000, Federal Register, with a July 1, 2000, implementation date. EDS might receive increased reimbursement for services under APCs, in sharp contrast to previous predictions of a 15% decrease in reimbursement. Instead of combining CPT and ICD-9-CM coding for clinic and emergency visits APCs, HCFA has assigned three APCs for the emergency department and a fourth APC for critical care. There is no APC for a medical screening exam, which could improve reimbursement. There is no separate APC for observation services, which means no additional payment will be given. You can continue to use your current charge structure and correlate present levels of service with the appropriate CPT visit level. You will have to "unbundle" visit levels to list nursing and physician procedures separately as specific line items. PMID- 11067335 TI - Know difference between APC coding systems. AB - You'll need to revise your chargemasters to include all of the evaluation and management CPT codes for emergency and clinic technical services to prepare for the Health Care Financing Administration's switch to ambulatory payment classifications for outpatient services. Physician- and hospital-generated CPT codes present different challenges. Physician CPT coding should never be performed on the higher-level visits without complete documentation of the higher level visits. A descriptive patient classification system (PCS) does not accurately portray the intensity of the visit, but the quantified PCS lists specific nursing tasks and assigns a point value to each one. PMID- 11067336 TI - Reports spotlight medication errors: make changes before tragedy strikes. AB - Three organizations recently published reports on medication errors--the Institute for Healthcare Improvement, the Institute of Medicine, and the Joint Commission on Accreditation of Healthcare Organizations. The ED is at higher risk for medication errors than other areas of the hospital because of extreme time pressure, high patient volumes, verbal orders, and multiple staff caring for patients. Medication errors often are caused by unclear labeling, lack of a check system for medication dosages, and similar drugs kept in close proximity to each other. Recommended strategies include a check system in which one nurse prepares a dose and another nurse reviews it, and avoiding look-alike and sound-alike medications. PMID- 11067337 TI - Abandoned infants can be brought to ED. AB - To reduce the number of abandoned newborns, a state law was passed in Texas that allows mothers to abandon infants anonymously in the ED and avoid criminal prosecution. Similar legislation has been introduced in Delaware, Alabama, California, Minnesota, and Georgia. Although the law aims to ensure children aren't abandoned in unsafe places, ED experts question whether it will be effective. Once it is determined a child is abandoned, the burden is on the ED staff to perform a medical evaluation with no background information. ED staff should educate pregnant women and young mothers about community resources in times of domestic crisis. PMID- 11067338 TI - Physician/patient rapport concerns administrators. AB - An ED staffing survey of 588 hospital administrators revealed widespread concerns about the quality of care in the ED and identified trends in staffing and operations. 40% of respondents reported concerns regarding physicians' rapport with patients. One in 10 administrators surveyed said they would not choose to go to their hospital's ED for treatment if they were seriously injured, and 20% indicated concerns about professional competence in the ED. Respondents indicated that more than 80% of ED physicians are involved in medical leadership of the hospital. PMID- 11067339 TI - States put case managers' clinical qualifications on trial. PMID- 11067340 TI - Is nursing a profession or merely a role? PMID- 11067341 TI - Don't waste dollars on useless automation. PMID- 11067342 TI - Practice teams boost pathway effort. PMID- 11067343 TI - How to avoid surprises with new software. PMID- 11067344 TI - How to manage clinical path variance data. PMID- 11067345 TI - The hospitalist model: proceed with caution. PMID- 11067346 TI - The case for hospitalists. PMID- 11067347 TI - Hospitalist care flourishes, amid physician concerns. PMID- 11067348 TI - Between a rock and a hard place. Conflicting demands on our health care system. Panel discussion. PMID- 11067349 TI - 25 years later: an unfinished revolution. Interview by Mark Hagland. PMID- 11067350 TI - Linked for better health. PMID- 11067351 TI - A prescription for physicians and their elderly patients. PMID- 11067352 TI - After midnight: post Y2K computer priorities. PMID- 11067353 TI - A workshop on better care for seniors. PMID- 11067354 TI - By the numbers. Satisfied seniors, stalled enrollment. PMID- 11067356 TI - Some medical groups need a little TLC of their own. PMID- 11067355 TI - The business of doctoring. PMID- 11067357 TI - New class action suits: what's at risk. PMID- 11067358 TI - Taking on diabetes. PMID- 11067359 TI - Meeting part way. A consensus (of sorts) on new health care reforms in California. Panel discussion. PMID- 11067360 TI - Getting it together: integrating disease management. PMID- 11067361 TI - New standards and measures for mental health care. PMID- 11067362 TI - Internet: the pharmacy of the future? PMID- 11067363 TI - Unraveling stress. PMID- 11067364 TI - By the numbers. The facts about diabetes. PMID- 11067365 TI - The 1999 numbers are in. Despite turmoil, acquisition prices were stable. PMID- 11067366 TI - Surviving the shake-up. PMID- 11067367 TI - 'Accredited' living facilities. Analyzing the costs and benefits of credentialing your ALF. PMID- 11067368 TI - Profiles. People who make a difference. PMID- 11067369 TI - Dos and doses. Techniques to prevent medication errors and inappropriate use. PMID- 11067370 TI - High court rebuffs challenge to survey enforcement process. PMID- 11067371 TI - Author weaves quite a story. PMID- 11067372 TI - Why are drug prices lower in Canada? PMID- 11067373 TI - Comparing the medical expenses of children with Medicaid and commercial insurance in an HMO. AB - BACKGROUND: In recent years, growing numbers of children with Medicaid have been enrolled in managed care plans nationwide. Yet, large, commercial managed care plans are increasingly discontinuing their participation in Medicaid because of low Medicaid payment rates. OBJECTIVE: To compare the healthcare utilization and costs of children with Medicaid and children with commercial insurance within the same health maintenance organization (HMO). STUDY DESIGN: Retrospective study using electronically captured cost and utilization data. PATIENTS AND METHODS: We compared the healthcare utilization and costs of children with Medicaid (n = 42,636) and children with commercial insurance (n = 159,651) who were members of the same large, nonprofit HMO at any time between January 1, 1995, and December 31, 1997. Medicaid children were grouped as income eligible, medically needy, and blind or disabled. RESULTS: The unadjusted costs of income-eligible, Medicaid insured children were not significantly different from those of commercially insured children. The medically needy were $25 per month more expensive than commercially insured children (P = 0.02), and the blind or disabled were $213 per month more expensive (P < .01). After adjusting for age and sex, income-eligible children were $5 per month more expensive than children with commercial insurance (P = .07), the medically needy were $20 per month more expensive (P = .02), and the blind or disabled were $216 per month more expensive (P < .01). CONCLUSIONS: The costs of income-eligible, Medicaid-insured children in this HMO were similar to those of commercially insured children, but the costs for the medically needy and the blind and disabled were substantially higher. PMID- 11067374 TI - Clinical and cost implications of new technologies for cervical cancer screening: the impact of test sensitivity. AB - OBJECTIVE: To compare the available techniques for cervical cancer screening, including several new technologies, using actual program utilization patterns. STUDY DESIGN: Longitudinal cohort model. PATIENTS AND METHODS: The model followed a cohort of 100,000 women who underwent screening from age 20 through 65 years. The model was run with a weighted average of screening intervals to model the actual utilization of the cervical cancer screening program in the United States. RESULTS: The model demonstrated that new technologies with significantly increased test sensitivity have the potential to reduce the number of cancers by 45% to 60% depending on the screening frequency in fully compliant populations. At screening intervals of 2 years or more, these new technologies had cost effectiveness ratios below $50,000 per life-year saved. Assuming existing utilization patterns, the model predicted there would be 13.2 cancers per year in the 100,000 women screened with the conventional Pap smear, and new technologies with increased test sensitivity could reduce the annual incidence to 9.5 cancers per 100,000 women screened. CONCLUSIONS: The model suggests that to achieve further dramatic reduction in cervical cancer mortality, significant improvements in test sensitivity, as reflected in the new screening technologies, may be the most realistic and cost-effective approach. PMID- 11067375 TI - High-risk population health management--achieving improved patient outcomes and near-term financial results. AB - OBJECTIVE: A managed care organization sought to achieve efficiencies in care delivery and cost savings by anticipating and better caring for its frail and least stable members. STUDY DESIGN: Time sequence case study of program intervention across an entire managed care population in its first year compared with the prior baseline year. PATIENTS AND METHODS: Key attributes of the intervention included predictive registries of at-risk members based on existing data, relentless focus on the high-risk group, an integrated clinical and psychosocial approach to assessments and are planning, a reengineered care management process, secured Internet applications enabling rapid implementation and broad connectivity, and population-based outcomes metrics derived from widely used measures of resource utilization and functional status. RESULTS: Concentrating on the highest-risk group, which averaged just 1.1% prevalence in the total membership, yielded bottom line results. When the year before program implementation (July 1997 through June 1998) was compared with the subsequent year, the total population's annualized commercial admission rate was reduced 5.3%, and seniors' was reduced 3.0%. A claims-paid analysis exclusively of the highest-risk group revealed that their efficiencies and savings overwhelmingly contributed to the membershipwide effect. This subgroup's costs dropped 35.7% from preprogram levels of $2590 per member per month (excluding pharmaceuticals). During the same time, patient-derived cross-sectional functional status rose 12.5%. CONCLUSIONS: A sharply focused, Internet-deployed case management strategy achieved economic and functional status results on a population basis and produced systemwide savings in its first year of implementation. PMID- 11067376 TI - Design and implementation of an inpatient disease management program. AB - OBJECTIVE: To describe the development and implementation of an inpatient disease management program. STUDY DESIGN: Prospective observational study. PATIENTS AND METHODS: On the basis of opportunities for improving quality or efficiency of inpatient and emergency department care, 4 diagnoses, including congestive heart failure (CHF), gastrointestinal hemorrhage, community-acquired pneumonia and sickle-cell crisis were selected for implementation of a disease management program. For each diagnosis, a task force assembled a disease management team led by a "physician champion" and nurse care manager and identified opportunities for improvement through medical literature review and interviews with caregivers. A limited number of disease-specific guidelines and corresponding interventions were selected with consensus of the team and disseminated to caregivers. Physician and nurse team leaders were actively involved in patient care to facilitate adherence to guidelines. RESULTS: For quarter 2 to 4 of 1997, there were improvements in angiotensin-converting enzyme inhibitor use, daily weight compliance, assessment of left ventricular function, hospital costs, and length of stay for care-managed patients with CHF. Differences in utilization-related outcomes persisted even after adjustment for severity of illness. For the other 3 diagnoses, the observational period was shorter (quarter 4 only), and hence preliminary data showed similar hospital costs and length of stay for care managed and noncare-managed patients. CONCLUSIONS: An interdisciplinary approach to inpatient disease management resulted in substantial improvements in both quality and efficiency of care for patients with CHF. Additional data are needed to determine the program's impact on outcomes of other targeted diagnoses. PMID- 11067377 TI - Managing drug costs: the perception of managed care pharmacy directors. AB - OBJECTIVE: To examine the perceptions of health plan pharmacy directors about drug costs and utilization drivers, interventions the plans use to control drug expenditures, and strategies considered necessary to permit continued provision of a comprehensive drug benefit. STUDY DESIGN/METHODS: A multipart survey developed and mailed to 500 pharmacy directors of managed care organizations across the country. RESULTS: The survey respondents (response rate = 18%) represented managed care health plans in the following percentages: 49% of respondents were from network/independent practice associations; mixed-model health maintenance organizations (HMOs), 20%; group HMOs, 15%; and staff-model HMOs and network/preferred provider organizations, 8% each. Drug mix and utilization were reported to be the primary drivers of drug expenditures. Half the respondents rated inflation as a somewhat strong cost driver. Interventions the health plans use to control drug expenditures include formularies, generic substitution, preauthorization, manufacturers' rebates, drug benefit design, physician profiling, target drug programs, academic detailing, and tiered copays. With the exception of formulary use, generic substitution, and manufacturers' rebates, which all the plans have instituted, the types of interventions used by the different model types vary widely. More than half the pharmacy directors reported generic substitution, drug benefit design, and differential copays as very effective interventions used to control drug costs. CONCLUSIONS: The majority of pharmacy directors predict continued double-digit increases in drug expenditures over both the short term and the long term. Of the respondents, 91% reported that additional limits and/or exclusions to the benefit design would be necessary to control these increases. To continue providing a comprehensive drug benefit, 54% indicated that they would have to achieve sufficient cost savings in other areas to offset increases in drug costs. PMID- 11067378 TI - Interactive voice response systems in the diagnosis and management of chronic disease. AB - OBJECTIVE: To assess the feasibility, reliability, validity, and potential clinical impact of interactive voice response (IVR) systems in the diagnosis and management of chronic disease. STUDY DESIGN: Literature review. RESULTS: Interactive voice response assessment systems have been implemented in the treatment of patients with chronic health problems such as heart failure, diabetes, hypertension, and mental health disorders. The information patients report during IVR assessments is at least as reliable as information obtained via structured clinical interviews or medical record reviews. Patients often are more inclined to report health problems to an IVR system than directly to a clinician. The few outcome evaluations of IVR-supported chronic illness management services indicate that they can have moderate impacts on some health and health behavior outcomes. CONCLUSIONS: Future research should evaluate the extent to which IVR assessment data can improve the prediction of clinical problems over and above what is possible using data usually available to primary care providers. Studies also should evaluate the outcomes of IVR-supported chronic disease management and the use of IVR assessments to measure variation in patient-centered treatment outcomes. PMID- 11067379 TI - Mandated managed care for blind and disabled Medicaid beneficiaries in a county organized health system: implementation challenges and access issues. AB - OBJECTIVES: The challenges of Medicaid managed care organizations that serve blind and disabled members are reviewed. Beneficiary satisfaction and access to care are assessed, and access problems are identified and resolved or minimized to the greatest degree possible. STUDY DESIGN: Formative evaluation consisting of a mailed survey and follow-up telephone outreach contacts. PATIENTS AND METHODS: A written survey was sent to more than 18,000 Supplemental Security Income (SSI) beneficiary members who were blind or disabled, with 5574 recipients responding. Of these, 1981 members identified issues that warranted 2106 follow-up telephone calls. RESULTS: Survey responses showed that beneficiaries had limited experience with managed care and were generally satisfied with access to primary care. The healthcare system used the study findings to develop focused training programs and materials, to initiate a special needs liaison program, and to revise guidelines to simplify and standardize authorization procedures for obtaining medical supplies and repairing equipment. CONCLUSIONS: Factors found to be associated with the success of a Medicaid managed care program serving blind and disabled beneficiaries include educating the members and providers for better understanding of managed care, incorporating collaborative service improvement activities, and documenting trends. PMID- 11067380 TI - Improving the sensitivity of cervical cytology: what are the issues? PMID- 11067381 TI - Error reporting: even if it doesn't seem broken, it still might need fixing. PMID- 11067382 TI - Hip replacement data lead to efficiency. PMID- 11067383 TI - Following the path to improved outcomes. PMID- 11067384 TI - Stats & facts. A closer look at third-party administrators. PMID- 11067385 TI - Everything is connected to the Net. PMID- 11067386 TI - The effect of HMOs on hospital capacity, 1982-1996. AB - Hospital capacity has declined in recent years and is forecast to decline further. The objective of this study is to determine if hospital capacity has declined more rapidly in metropolitan statistical areas (MSAs) with high HMO market penetration compared with MSAs with low HMO market penetration in the period from 1982 to 1996. The findings presented in this study are that greater reductions in beds per capita occur in MSAs with greater HMO penetration, but the magnitude of the differences is small. Reductions in other measures of hospital capacity--hospital closures, beds per hospital, and occupancy--are not consistently associated with HMO market penetration. Bed capacity occurs at roughly similar rates in all MSAs. PMID- 11067387 TI - Training the next generation of managed care pharmacists. PMID- 11067388 TI - Evidence-based practices and the National Institute for Clinical Excellence. PMID- 11067389 TI - Values and the application of information technology. Interview by Ross Martin. PMID- 11067390 TI - E-mail marketing grows up: a primer for the managed care industry. AB - Managed care plans are jumping onto the electronic marketing bandwagon in a big way, taking advantage of not only the basic E-mail system but also expanding on that medium and developing creative vehicles to send the health plan's message. In this article, the author describes how E-mail technology is being used to hone the marketing edge in MCOs. PMID- 11067391 TI - A pharmacoeconomic evaluation of two new products for the treatment of overactive bladder. AB - The objective of this study is to evaluate the cost effectiveness of two new treatments for overactive bladder: once-daily controlled-release oxybutynin, and twice-daily tolterodine, with a comparison with oxybutynin immediate release. Also estimated are the potential cost savings to a health plan budget resulting from increased utilization of the most cost-effective treatment. The design is a decision-tree model based on clinical trial data and expert panel estimates with a six-month time horizon conducted from a payer perspective. The primary outcome measure used in the analysis was treatment success, with success defined as zero incontinence episodes per week. A secondary outcome measure was the expected number of continent days. As first-line therapy, controlled-release oxybutynin is the most cost-effective treatment as measured by expected cost per success and expected cost per continent days. Controlled-release, once-daily oxybutynin yielded the highest expected success rate and the highest number of expected continent days. The expected cost of treatment with controlled-release oxybutynin was lower than tolterodine and equivalent to immediate-release oxybutynin. Increased utilization of controlled-release oxybutynin results in an estimated saving of $0.007 to $0.026 per member per month for a hypothetical HMO. The model was robust, incorporating all assumptions based on univariate and multivariate sensitivity analysis. Initiating treatment with controlled-release oxybutynin is the most cost-effective approach to treatment for overactive bladder. PMID- 11067392 TI - The art of premium setting. PMID- 11067393 TI - The implications of a Medicare prescription drug benefit. PMID- 11067394 TI - B2B: do it better, cheaper, faster (on-line). PMID- 11067395 TI - Ischemic heart disease. PMID- 11067396 TI - Small businesses need affordable health coverage. PMID- 11067398 TI - Pharmacy management and three-tier copays: cost management or cost shifting? PMID- 11067397 TI - Beta-blocker use in patients with acute myocardial infarction treated by hospitalists. AB - It is well recognized that the administration of beta blockers to patients after myocardial infarction improves survival. This retrospective cohort and prospective study sought to define the usage of a large hospitalist group and enhance this usage by education and the utilization of a uniform discharge summary. All patients with a discharge diagnosis of acute myocardial infarction were included for analysis. The use of beta blockers by the hospitalist group was initially collected retrospectively and compared with two large cohorts. The data were presented to the hospitalist group. Prospective data collection then commenced. Retrospective analysis of the use of beta blockers showed a rate of 68% as compared with 21% and 34% in two large cohorts (P < .0001). After data were reviewed and conference occurred, prospective use of beta blockers increased to 90% (P < .0005). Patients with myocardial infarction were extremely likely to be treated with beta blockers by this hospitalist group. Review of previous usage and review of contraindications along with the use of a uniform discharge summary resulted in a significant increase in the use of these life-saving drugs. PMID- 11067399 TI - The role of innovation in restructuring health care delivery. PMID- 11067400 TI - Getting gung ho on guidelines. PMID- 11067401 TI - The use of antiplatelet agents in cardiac disease. AB - In this clinical review, the authors focus on the process of platelet activation and thrombosis in acute and chronic coronary syndromes. New antithrombotic therapies have emerged for various cardiovascular diseases, based on the evolving concept of vascular injury and platelets and the knowledge of the pathophysiology of the underlying disease states. An understanding of these mechanisms is crucial to therapeutic decision making with antiplatelet regimens, which are also discussed in detail. PMID- 11067402 TI - A textbook example of how to fail at risk contracting. PMID- 11067403 TI - Will capitation contracts soon include patient safety standards? AB - The rare delivery systems that balance clinical excellence with cost containment have been waiting patiently for purchasers to reward them with preferential capitation rates or higher member volume. A new coalition of employers plans to do just that. PMID- 11067404 TI - Evidence-based medicine helps providers offer cost-effective care. PMID- 11067405 TI - Health system treads water on these rates, but looks forward to better days ahead. PMID- 11067406 TI - Consider these strategies for managing rising drug costs. PMID- 11067407 TI - Physicians charge California HMOs with extortion, racketeering. PMID- 11067408 TI - Using a quality-based incentive system to reward efficient behavioral health providers. PMID- 11067409 TI - Get providers the data they need to manage risk successfully. PMID- 11067410 TI - Highmark delays episode payments for specialists. AB - In the wake of opposition from participating specialty physicians and reports of problems with similar systems around the country, Highmark Blue Cross and Blue Shield says it will wait until early 2001 to implement changes in the way it pays three types of specialty physicians for treating its Medicare HMO members. PMID- 11067411 TI - Texas MDs backpedal on initial praise for Aetna settlement. PMID- 11067412 TI - Interview with Elizabeth Calhoun, M.H.A. student, and Elizabeth Wennar, D.H.A. candidate, Medical University of South Carolina. Interview by James A. Johnson. PMID- 11067413 TI - Restructuring employment relationships between healthcare organizations and primary care physicians. PMID- 11067414 TI - Making the tough calls. PMID- 11067415 TI - Managing information resources: a study of ten healthcare organizations. AB - This article presents the results of information technology management audits conducted by senior executives at ten healthcare organizations. The audits evaluated how well the following seven information technology management responsibilities were carried out: (1) strategic information systems planning; (2) employment of a user focus in system development; (3) recruiting of competent personnel; (4) information systems integration; (5) protection of information security and confidentiality; (6) employment of effective project management in system development; and (7) post-implementation evaluation of information systems. The audit results suggest that most of these responsibilities are being met to a considerable extent by a majority of the organizations studied. However, substantial variation across organizations was noted. Executives participating in the study were able to define areas in which the management of information resources in their organizations was in need of attention. The audit process encourages senior management to provide the leadership required to ensure that information technology is used to maximum advantage. PMID- 11067416 TI - The diffusion of decision support systems in healthcare: are we there yet? AB - Clinical decision support (CDS) systems, with the potential to minimize practice variation and improve patient care, have begun to surface throughout the healthcare industry. This study reviews historic patterns of information technology (IT) in healthcare, analyzes barriers and enabling factors, and draws three lessons. First, the widespread adoption of clinical IT, including CDS systems, depends on having the right organizational and individual financial incentives in place. Second, although CDS systems and clinical IT in general are powerful tools that can be used to support the practice of medicine, they alone cannot redefine the workflow or processes within the profession. Healthcare managers counting on technology to restructure or monitor clinicians' work patterns are likely to encounter substantial resistance to CDS systems, even those that generate valuable information. Third, while the pace of implementing IT systems in healthcare has lagged behind that of other industries, many of the obstacles are gradually diminishing. However, several factors continue to inhibit their widespread diffusion, including the organizational turmoil created by large numbers of mergers and acquisitions, and the lack of uniform data standards. PMID- 11067417 TI - Disease management: old wine in new bottles? AB - Disease management is a holistic, patient-focused approach to the treatment of disease across the spectrum of healthcare delivery. In its current form, disease management was created in response to the societal and economic burden that chronic illness contributes. There has recently been rapid growth in the development of disease management programs and sponsors are widespread within the industry, with the largest increase in independent vendors. Although growth has been substantial, the hurdles these programs have encountered have kept them from reaching their full potential. The challenges that exist include clinical, financial, and regulatory issues, and these challenges have significant meaning to healthcare managers. In deciding whether to develop or enhance programs, executives must consider their capability of outcomes measurement, their provider relationships, and the arrangements for program implementation. Ultimately, if programs provide improved health and quality of life for participants, cost savings will follow. PMID- 11067418 TI - Mission and organizational performance in the healthcare industry. AB - Healthcare is a rapidly evolving industry where firms face constantly changing conditions and an ever-increasing demand for services. As competition in this sector continues to grow, managers must assess their organizations and develop methods to improve firm performance and productivity. Successful managers are constantly searching for tools that will motivate their employees to perform at the highest possible level. An increasing amount of literature points to the mission statement as a valuable tool for managers to use to improve organizational performance and increase employee motivation. This article examines the key elements of the organizational mission statement and discusses their significance in relation to firm performance in the healthcare industry. First, the elements of a mission statement will be identified and the relationship of these elements to both organizational success and employee motivation will be discussed. Second, the general mission development process and its most critical features will be considered. Third, specific mission statements from a sample of healthcare organizations will be identified and analyzed using an integrated analytical framework based on the literature. Finally, the significance of the material presented will be considered followed by suggestions for future mission statement development and evaluation. PMID- 11067419 TI - Interview with Tomer Loiter, founder and CEO of Med on Web, Inc. Interview by James A. Johnson. PMID- 11067420 TI - Optimal strategies for health systems and physician practices with limited financial resources. PMID- 11067421 TI - The "business"--or "public service"--of healthcare. PMID- 11067422 TI - Creating a vision for the twenty-first century healthcare organization. AB - Management approaches used by healthcare organizations have often lagged behind other businesses in more competitive industries. Companies operating in such dynamic environments have found that to cope with the rapid pace of change they must have an articulated understanding of their organization's capabilities and consensus on where the organization is headed based on predictions about the future operating environment. This statement of identity and strategic direction takes the form of a vision statement that serves as the compass for the organization's decisions for a five- to ten-year period. This article discusses the importance of vision statements in tomorrow's healthcare organizations, presents an overview of future scenarios that may provide context for organizational visions, and suggests a process for developing a vision statement. A case study is presented to illustrate how a vision statement is created. Following the guidelines presented in this article and reviewing the case study should assist healthcare executives and their boards in crafting better visions of their organizations' futures, developing more effective strategies to realize these visions, and adapting to more frequent and more significant change. PMID- 11067423 TI - A life cycle model of continuous clinical process innovation. AB - The changing healthcare environment has created a sense of urgency for continuous innovation in clinical care processes. Managers and clinicians are investing unprecedented funds and energy in the development of various clinical process innovations (CPI) such as clinical pathways, electronic workstations, and various forms of information technology. While increasing attention has been paid to the development of such initiatives, our understanding of how best to disseminate and ensure their use is limited. In this first of two articles dealing with the dissemination and use of CPI in integrated delivery systems, we present a "life cycle" model of the dissemination process and suggest opportunities for managing CPI. The management of CPI requires more than just an understanding of the factors that may facilitate or impede its implementation and use. Managers require an understanding of the actual process so that they can assess the specific implementation stage at which the organization is presently operating, and design appropriate interventions that can affect the process. A future article will identify the factors that facilitate and inhibit the process and suggest some intervention strategies. PMID- 11067424 TI - The timing of medical technology acquisition: strategic decision making in turbulent environments. AB - Healthcare decision makers and researchers have long been interested in the factors behind medical technology acquisition. The rate of environmental change in recent years has dramatically affected technology acquisition decision making in acute care hospitals. This study examines the relative role of decision-maker influence and environmental factors on the timing of MRI acquisition in hospitals operating in three western states with different levels of environmental uncertainty. The results suggest that the relative influence of decision makers and environmental factors on innovation acquisition timing varies depending on environmental turbulence, and that hospitals acquire new technology as one way of controlling the turbulence in their environments. PMID- 11067425 TI - Healing models for organizations: description, measurement, and outcomes. AB - Healthcare leaders are continually searching for ways to improve their ability to provide optimal healthcare services, be financially viable, and retain quality caregivers, often feeling like such goals are impossible to achieve in today's intensely competitive environment. Many healthcare leaders intuitively recognize the need for more humanistic models and the probable connection with positive patient outcomes and financial success but are hesitant to make significant changes in their organizations because of the lack of model descriptions or documented recognition of the clinical and financial advantages of humanistic models. This article describes a study that was developed in response to the increasing work in humanistic or healing environment models and the need for validation of the advantages of such models. The healthy organization model, a framework for healthcare organizations that incorporates humanistic healing values within the traditional structure, is presented as a result of the study. This model addresses the importance of optimal clinical services, financial performance, and staff satisfaction. The five research-based organizational components that form the framework are described, and key indicators of organizational effectiveness over a five-year period are presented. The resulting empirical data are strongly supportive of the healing model and reflect positive outcomes for the organization. PMID- 11067426 TI - Governance change for public hospitals. PMID- 11067427 TI - The "incredible shrinking Medicare program": Treasury numbers tell troublesome tale. PMID- 11067428 TI - House Budget Committee holds hearing on Medicare regulation. PMID- 11067429 TI - The Medicare prescription drug debate. PMID- 11067431 TI - The OIG's self-disclosure protocol. PMID- 11067432 TI - FTC and DOJ jointly propose antitrust guidelines for collaborations among competitors. PMID- 11067433 TI - A discussion of pharmaceutical industry practices viewed as illegal by the federal government. PMID- 11067434 TI - Innovative approaches to Medicare and commercial membership retention. PMID- 11067436 TI - Variations in care not related to capitation, study shows. PMID- 11067435 TI - Direct-to-consumer models offer important revenue opportunities. PMID- 11067437 TI - Data confirm importance of health status adjustment. PMID- 11067438 TI - Study shows increases in drug costs may be justified by improvements in outcomes. PMID- 11067440 TI - Use this tool to assess patient satisfaction rates. PMID- 11067439 TI - AHRQ offers robust new benchmarking tool--and it's free. PMID- 11067441 TI - Time for accountability: facing the tough questions will build public trust. PMID- 11067442 TI - Improving medication administration error reporting systems. Why do errors occur? AB - Monitoring medication administration errors (MAE) is often included as part of the hospital's risk management program. While observation of actual medication administration is the most accurate way to identify errors, hospitals typically rely on voluntary incident reporting processes. Although incident reporting systems are more economical than other methods of error detection, incident reporting can also be a time-consuming process depending on the complexity or "user-friendliness" of the reporting system. Accurate incident reporting systems are also dependent on the ability of the practitioner to: 1) recognize an error has actually occurred; 2) believe the error is significant enough to warrant reporting; and 3) overcome the embarrassment of having committed a MAE and the fear of punishment for reporting a mistake (either one's own or another's mistake). PMID- 11067443 TI - MedTeams: teamwork advances emergency department effectiveness and reduces medical errors. PMID- 11067444 TI - Developing a culture of patient safety at the VA. AB - Patient safety is a topic that has become prominent in the minds of many, both within and outside the healthcare field over the past several months. But in fact, literature in medical journals describing this topic goes back decades. However, studying these issues is only the first step towards developing useful and practical tools to address errors and does little to change the safety culture that underlies these systems. The VA has taken several steps towards a safety culture and the development and implementation of tools, such as: 1) error reporting mechanisms; 2) tools for root cause and corrective action; and 3) management tools (e.g., safety awards). PMID- 11067445 TI - A mindful infrastructure for increasing reliability. AB - Traditional analyses of adverse medical events and errors have focused on individuals. The search for a cause typically has stopped at the person closest to the accident who, it is determined after the fact, could have acted differently in a way that would have led to a different outcome. Traditional approaches have focused on people as unreliable components. But the new look at error has shifted its focus from individuals to the systems in which these individuals are situated. I want to add to this discussion by reporting on an analysis of non-medical organizations called "high reliability organizations" (or HROs) that incur similar temptations to blame individuals rather than systems, but have been successful in focusing attention on systems. The point of this discussion is to suggest that the ways in which HROs do this are instructive for medical organizations whose goal is fewer adverse events. PMID- 11067446 TI - Action to improve patient safety: "safety" prone health care systems. AB - The error prone health care system is complex, tightly coupled and hierarchical. Who's at fault when an error occurs? How do we keep patients safe and prevent errors in this error prone system? There will continue to be health care mistakes, it is inevitable in an error prone system but things can be done to increase patient safety. The communication between and among health care providers and patients that work toward building better relationship ties have demonstrated the potential for greater patient safety. In fact, starting from the discussion point of patient safety, rather than starting from error, has the most profound chance to benefit patients. An overview of efforts to increase patient safety through research and clinical practice are discussed. Ironically, examples of errors in health care have caught the attention of the American public. In the long run, patient safety must be the intrinsic cause for improvement. Many errors in health care are unknown and the total number may be unknowable. A well-known study from Harvard reported that about 4 percent of hospitalized patients had iatrogenic injuries; 13 percent of those were fatal (Leape et al, 1991). The principle investigator in that study, Dr. Lucien Leape, said "Errors are system flaws, not character flaws". In 95 percent of the cases, errors are not the result of carelessness or lack of concern. The worse errors are sometimes made by the best doctors and nurses (Leape et al, 1991). Although technology is helping in some ways, it is also causing a growing risk of new unexpected adverse events. This is a problem that must be addressed. Even though not a popular problem in health care, if not critically tackled, it will get worse in the future. This article examines: why this problem needs to be addressed, what has been done so far, and the major components of health care, systems, technology, and humans, that make it error prone and complex. This article will also examine these three areas of interest where mistakes are made. PMID- 11067447 TI - Disabling conditions. PMID- 11067448 TI - ALS for the BLS provider. PMID- 11067449 TI - A Yankee in England. An American paramedic explores EMS in the UK. PMID- 11067450 TI - Vive la difference! PMID- 11067451 TI - "Man acting strangely". PMID- 11067452 TI - Dermal diagnosis. PMID- 11067453 TI - Prehospital management of neck trauma. PMID- 11067454 TI - Maxillofacial trauma. PMID- 11067455 TI - A matter of time. Part II: Is EMS ready for domestic terrorism? PMID- 11067456 TI - EMS incident management: operational communications. PMID- 11067457 TI - All clear? Researchers are currently investigating whether EMS providers should clear potential spinal injuries in the field. PMID- 11067458 TI - Effective state EMS lobbying: it's all in the plan. AB - Any piece of legislation needs grass roots support. If local legislators are not hearing from local EMS systems, the bill will have an uphill battle in the legislative assembly. Bill sponsors and key supporters need to be regularly informed of the bill's status. Remember the KISS theory--all testimony and communication between local EMS lobbyists and legislators needs to be kept at an understandable level. Finally, remember to never quit your lobbying until the Governor signs the bill into law. PMID- 11067459 TI - Parents: the "secondary patients". PMID- 11067460 TI - Patient assessment skills. PMID- 11067461 TI - Heatwave. Prehospital management of heat-related conditions. PMID- 11067462 TI - Writing run reports: an opportunity for learning and professional growth. PMID- 11067463 TI - A is for airway: an oral intubation primer. PMID- 11067464 TI - Leaving the ivory tower. PMID- 11067465 TI - Unwanted advances: sexual harassment in the workplace. PMID- 11067466 TI - A secondary survey: other laws and rules controlling EMS. PMID- 11067467 TI - EMS in the Rhineland. PMID- 11067468 TI - Sizzling special events. Does air temperature affect patient encounters at mass gatherings? PMID- 11067469 TI - All change in the Windy City. Interview by Mary Nordberg. PMID- 11067470 TI - Are you guilty of unsafe documentation? PMID- 11067471 TI - Basilar skull fracture. PMID- 11067472 TI - Workplace violence prevention: one hospital's approach. PMID- 11067473 TI - An interview with: Richard D. Maurer on the new roles and responsibilities of security directors. PMID- 11067474 TI - How two hospitals reacted to parking lot crimes. PMID- 11067475 TI - Taking advantage of security opportunities in new hospitals--large and small. AB - Although in many parts of the country the trend is for hospitals to merge, consolidate, or even shut down due to financial constraints and increased competition from managed care, in other areas new hospitals are being built--and with more advanced security systems for protecting patients, visitors, and staff. In this report, we'll give details on the security planning involved in the building of four new facilities, ranging in size from 56 to 400 beds, each in a different region of the country. PMID- 11067476 TI - The ins and outs of complying with hiring/pre-employment screening laws. PMID- 11067477 TI - Health Management Awards. Peak practice. PMID- 11067478 TI - Efficiency. Measured response. AB - An analysis of the activity of 75 acute hospitals over the period 1991-96 using data envelopment analysis shows that, while overall productivity increased, the efficiency of individual hospitals did not. A small decrease in the efficiency of individual hospitals was found in the last four years studied. An analysis of quality of care over the same period suggests that gains in volume of services may have been at the expense of quality of care. The results suggest that incentives for increasing hospital efficiency have a one-off impact rather than a sustained effect. PMID- 11067479 TI - Clinical priorities. Points make prizes. AB - Assessing patients for non-urgent surgery according to a numeric priority system has rationalised waiting times in three specialties in a pilot project. Traditional scoring systems result in little relation between urgency and waiting times. Enlisting doctors' support for a scoring system takes time. PMID- 11067480 TI - Primary care. Communal living. AB - From this month, 80 per cent of health authorities will have at least one personal medical services pilot scheme in their area. The extension of the schemes raises concerns about equity and management issues for HAs. An HA where schemes will cover almost a third of the population has had to adopt an off-the peg approach to development. PMID- 11067481 TI - Chief executives. Scapegoats in the wilderness. AB - At least 40 chief executives have had their contracts terminated early in the past five years. Very few, if any, have gone through a disciplinary procedure. The most commonly stated reasons have been poor financial control and disagreements with chairs. This trend represents a considerable waste of talent. The experience required of a chief executive cannot be acquired overnight. The 'sudden death' of people at the top corrupts organisations and deprives them of continuity. PMID- 11067482 TI - If I may be so bold.... PMID- 11067483 TI - Primary care trusts. Birds of a feather. AB - A trust which provides acute, community, mental health and learning disability services has been active in supporting the development of four primary care trusts in its area. All will start this year. Initially there were suspicions about an organisation voting for its own demise. It is important to focus on what is best for the clinical services rather than protecting the status quo. PMID- 11067484 TI - Primary care. Super groupers. PMID- 11067485 TI - Genetics regulation. Central line on the tubes. PMID- 11067486 TI - Data briefing. Hypothecated taxes. PMID- 11067487 TI - Telemedicine. As not seen on TV. PMID- 11067488 TI - US healthcare. Give and let live. PMID- 11067489 TI - Mental health. Facing hard truths. AB - A survey of 104 mental health units found that half did not have policies on treatment of detained patients from black and ethnic minority groups. Almost three-quarters had no policy for dealing with racial harassment of black and ethnic-minority patients. 11 percent of patients whose notes were examined had reported racial harassment. PMID- 11067490 TI - Mental health. On the case. PMID- 11067491 TI - PFI (private finance initiative) and IT (information technology) procurement. Going round in circulars. PMID- 11067492 TI - GPnet. Go with the flow. PMID- 11067493 TI - Data confidentiality. Drug habits in the dock. PMID- 11067494 TI - DECT (digitally enhanced cordless) telephony. Outwardly mobile. PMID- 11067495 TI - E-prescribing. All dosed up. PMID- 11067496 TI - Electronic blood matching. Antibodies of evidence. PMID- 11067497 TI - Reconfiguring services. Slow motion. AB - A scheme to reconfigure mental health services took 10 years from agreement in principle to the opening of the new unit. A plan to finance it under the private finance initiative had to be abandoned when the private investor pulled out. The original design was substantially altered in response to service users' requests. The trust allocated 1 per cent of the budget for artworks in the new unit. PMID- 11067498 TI - Mental health. In the frame. PMID- 11067499 TI - The comfort of small things. PMID- 11067500 TI - On the evidence. Type 2 diabetes. PMID- 11067501 TI - Mergers. Turn of the screw. AB - Managing the process of a merger with staff support and job counselling reduces stress. But the experience is not pain-free. The expectation that senior staff would be found other jobs in the NHS proved unrealistic. From a total staff of 6,000 there were 38 redundancies, of which 13 were voluntary. Early retirement and redundancy costs were higher than expected. Staff still show signs of distress a year after the merger took place. PMID- 11067502 TI - Mergers. Question of attitude. AB - Research with staff in two organisations involved in a merger revealed some convergence of cultures one year after the merger. But staff from the smaller organisation perceived some losses. Staff from both organisations felt poorly rewarded for their work. Staff from the smaller organisation perceived relationships in the organisation more positively than those in the larger one, before and after the merger. Staff saw their employers as very concerned with safety before and after the merger. PMID- 11067503 TI - Getting the wind up. PMID- 11067504 TI - Ethnic minority health. Lip service. PMID- 11067505 TI - Data briefing. Income distribution. PMID- 11067506 TI - Chief executives' pay. Topsy turvy. PMID- 11067507 TI - Nursing home regulation. Shadowlands. AB - The healthcare of 200,000 people living in nursing homes will not be regulated by the Commission for Health Improvement or by the proposed healthcare division of the National Care Standards Commission under current proposals. This is despite the fact that patients in nursing homes are highly dependent and many would previously have been cared for by the NHS. It is not acceptable that the current NHS quality initiatives should ignore this sector. PMID- 11067508 TI - Behind closed doors. PMID- 11067509 TI - Mental health. Forbidden fruit. PMID- 11067510 TI - New year, new regs. Time for managers to ... get up to speed! PMID- 11067511 TI - Aftermath ... hospitals are on the front lines after acts of terrorism. Are you prepared? PMID- 11067512 TI - Ice follies. This safety checklist puts the win in winter. PMID- 11067513 TI - Facility profile. Designers deliver spacious maternity center. Utah Valley Regional Medical Center, Provo, Utah. PMID- 11067514 TI - Facilities of the future. [New designs put patients first]. PMID- 11067515 TI - From the top--10 rules for outsourcing linen service. PMID- 11067516 TI - Signs of the times. New concepts for 21st century wayfinding. PMID- 11067517 TI - In the spotlight. NFPA meeting highlights health care. PMID- 11067518 TI - Heavy metal. Mercury is dropping off the charts--but not fast enough. PMID- 11067519 TI - Parents' reliance on physicians' advice might constitute contributory negligence. Alexander v. University of Pittsburgh Medical Center System (UPMCS). PMID- 11067520 TI - Erosion of employment-at-will doctrine continues. Deerman v. Beverly California Corp. PMID- 11067521 TI - Unfinished business among the aging and those who love them. PMID- 11067522 TI - Clinical pastoral education with students from other cultures: the role of the supervisor. PMID- 11067523 TI - Agape and moral meaning in pastoral counseling. PMID- 11067524 TI - The near death experience: observations and reflections from a retired chaplain. AB - In this article I have attempted to draw connections between the near-death experience, biblical spirituality, and the ministry of the spiritual caregiver. Also, I have described the near-death phenomena and given particular emphasis to the dying and death processes, including the offering of guidelines for approaching death. In my ministry and studies, I have come to believe that the near-death experience prepares us not only to die but to live life to the fullest until we say good-bye. PMID- 11067525 TI - When the caregiver needs care: a review of an interdisciplinary team response to stressful events in a tertiary care setting. PMID- 11067526 TI - Spiritual care in the hospital: who requests it? Who needs it? PMID- 11067527 TI - Delight: a discourse upon the spiritual function of the psyche. PMID- 11067528 TI - One of our own. PMID- 11067529 TI - Four days in the school of pain. PMID- 11067530 TI - Entering the minds of the mentally ill. PMID- 11067531 TI - Soul care: spiritual CPR. PMID- 11067532 TI - Verbatim: J B-A. PMID- 11067533 TI - HCFA issues proposed rule on uniform Medicare payment policy. PMID- 11067534 TI - Effectively motivating employees and accepting student interns. PMID- 11067535 TI - Drug screening: interactions, pitfalls, and techniques. PMID- 11067536 TI - Controlling blood culture contamination rates. PMID- 11067537 TI - Is it time for a career renewal? PMID- 11067538 TI - HIPAA compliance could cost dearly. PMID- 11067539 TI - Management competencies for the POL (physician office laboratory): Part 1, Self assessment. PMID- 11067540 TI - The LIS as process manager. Save money and reduce tech fatigue. PMID- 11067541 TI - Laboratory evaluation of discolored urine. When is it hematuria? PMID- 11067542 TI - 2000 Executive War College offers winning business strategies for the lab. PMID- 11067543 TI - What you need to know about latex allergy. PMID- 11067544 TI - New weapon for feds. PMID- 11067545 TI - $745 million and far to go. PMID- 11067546 TI - Kaiser's road to fiscal fitness. PMID- 11067547 TI - A voice for patients. PMID- 11067548 TI - In Beltway, all eyes are on drug benefit. PMID- 11067549 TI - No place for politics. PMID- 11067550 TI - Pricier, less plentiful. For many providers, gone are the days of free-flowing capital. PMID- 11067551 TI - Debate over charity care heats up Miami market. PMID- 11067552 TI - More lessons from AHERF. PMID- 11067553 TI - Web survey. April survey results: a look at medical errors. PMID- 11067554 TI - How ORs decide when to count instruments. PMID- 11067555 TI - E-commerce to rattle supply chain for ORs. PMID- 11067556 TI - Sterilization & infection control. The 'dating game' 25 years later. PMID- 11067557 TI - Big price rise in hip, knee implants noted. PMID- 11067558 TI - Infection control during remodel. PMID- 11067559 TI - Ambulatory surgery. Safe harbor rules raise questions. PMID- 11067560 TI - Ambulatory surgery. Accreditation queries answered. PMID- 11067561 TI - FDA steps up reg activity on reuse. PMID- 11067562 TI - HCFA to remove rule on CRNA supervision. PMID- 11067563 TI - JCAHO names measures for surgery. PMID- 11067564 TI - Managed care coverage of emergency services reemphasized. PMID- 11067565 TI - Patient satisfaction and billing: managing the last impression. PMID- 11067566 TI - Characteristics of elderly home health care users: data from the 1996 National Home and Hospice Care Survey. AB - OBJECTIVE: This report presents demographic characteristics, service utilization, and primary admission diagnoses of elderly users of home health care services. Included are home health care services used by both current and discharged clients (called patients). These services are provided by home health care agencies and hospices. The focus of the report is on services used by both current patients and discharges aged 65 years and over. METHODS: The data used for this report are from the National Center for Health Statistics 1996 National Home and Hospice Care Survey's (NHHCS) sample of current patients and discharges. The 1996 NHHCS is the fourth survey of home health care agencies and hospices and their current patients and discharges. RESULTS: The overall results of the survey indicate that, as in previous years, the elderly current patients and discharges were predominantly women, 75-84 years old, white, non-Hispanic, widowed, and most often lived in a private residence with members of their family. For elderly men and women, the most commonly used home health care service was skilled nursing services and the primary admission diagnosis was diseases of the circulatory system, including heart disease. PMID- 11067567 TI - Special issue on health promotion among the elderly. PMID- 11067568 TI - Preventive health behaviors and mammography use among urban older women. AB - A cross-sectional survey of 610 low income women between the ages of 60 and 84 who attended community meal sites in Los Angeles was conducted to determine health behaviors associated with mammography use among urban community dwellers. Preventive practices that require women to take an active role and recurrent participation were positively associated with a current mammography, while services that are clinician-initiated were associated with ever having a mammography. PMID- 11067569 TI - Is hypercholesterolemia a risk factor and should it be treated in the elderly? AB - PURPOSE: The 1993 National Cholesterol Education Program guidelines recommend cholesterol screening for elderly patients with and without known coronary heart disease. This review summarizes clinical trial evidence from the medical literature that addresses cholesterol treatment in the elderly. DATA SOURCES: References were obtained from a MEDLINE search, bibliographies, metaanalyses, and review articles. STUDY INCLUSION AND EXCLUSION CRITERIA: Randomized, controlled clinical trials, including all lipid intervention trials with elderly participants or subgroup analyses of the elderly designed to measure major cardiovascular disease endpoints, were selected. DATA EXTRACTION METHODS: A MEDLINE search of all clinical trials using key search terms yielded 1360 references. Journal titles and abstracts were reviewed for all references by one of us (K.M.H.). A full journal review was undertaken for 41 references to clinical trials. Five clinical trials fulfilled all criteria and represented unique data. DATA SYNTHESIS: A MEDLINE search (from 1966 to January 2000) and bibliography reviews yielded five important clinical trials with analyses of elderly participants. Data are presented in text form and a summary table. MAJOR CONCLUSIONS: Clinical trial evidence supports treating hyperlipidemia in elderly persons for secondary prevention of coronary heart disease. Evidence from four secondary prevention trials demonstrated that major coronary heart disease risk decreased by 25% to 30% in elderly subjects treated for 5 years. Unanswered questions include cholesterol treatment for primary prevention in the elderly, gender effect, and benefit of treatment in persons older than 70. PMID- 11067570 TI - Cigarette smoking among the elderly: disease consequences and the benefits of cessation. AB - The disease consequence of smoking occurs disproportionately among the elderly because of the long duration of cumulative injury or change that underlies the bulk of tobacco-caused disease. Older smokers are less likely than younger smokers to attempt quitting, but they are more likely to be successful in the attempts that they do make to quit. Excess absolute rates of disease incidence and mortality due to smoking increase steadily with increasing age and duration of smoking, and there is little evidence to suggest that the disease consequences of smoking diminish among the elderly. Although cardiovascular disease is the most common cause of excess mortality among younger smokers, lung cancer is the largest cause of excess smoking-related mortality over the age of 60 years; and at older ages the excess death rate from chronic obstructive lung disease equals that for cardiovascular disease. Because of the dramatic increases in smoking related excess mortality with advancing age, approximately 70% of the 400,000 or more deaths occur among those over age 60 years. The benefits of cessation are proportionately somewhat less among the elderly and may manifest more slowly than among younger smokers, but cessation remains the most effective way of altering smoking-induced disease risks at all ages, including those over the age of 60 years. PMID- 11067571 TI - Health promoting effects of friends and family on health outcomes in older adults. AB - OBJECTIVE: To highlight the significant impact of social relationships on health and illness and suggest implications of these effects for health promotion efforts among older adults. DATA SOURCES: Published studies on social relationships and health (or health behaviors) for the period 1970-1998 were identified through MEDLINE by using the key words social relationships, social support, and health, as well as review of health-related journals such as the American Journal of Epidemiology, Annals of Epidemiology, American Journal of Public Health, Journal of Health and Social Behavior, Social Science and Medicine, and the Journals of Gerontology. STUDY SELECTION: Major published original research was considered. Where published research was too extensive for full discussion of all studies, preference was given to studies focusing on older adults and those using stronger methodology (i.e., representative samples, longitudinal data, or multivariate analyses controlling for potential confounders). DATA EXTRACTION: Reported findings were organized in terms of three major categories: (1) results related to major health outcomes such as mortality, CHD, and depression; (2) findings related to health behaviors; and (3) findings related to potential biological pathways for observed health effects of social relationships. DATA SYNTHESIS: Protective effects of social integration with respect to mortality risk among older adults are the most thoroughly documented, although protective effects have also been documented with respect to risks for mental and physical health outcomes and for better recovery after disease onset. There is also now a growing awareness of the potential for negative health effects from social relationships that are characterized by more negative patterns of critical and/or demanding interactions, including increased risks for depression and angina. Biological pathways are suggested by evidence that more negative social interactions are associated with physiological profiles characterized by elevated stress hormones, increased cardiovascular activity, and depressed immune function, whereas more positive, supportive social interactions are associated with the opposite profile. CONCLUSIONS: Available data clearly indicate that social relationships have the potential for both health promoting and health damaging effects in older adults, and that there are biologically plausible pathways for these effects. Such evidence suggests that aspects of the social environment could play an important role in future health promotion efforts for older adults, although careful consideration of both potentially positive as well as negative social influences is needed. PMID- 11067572 TI - Health promotion for older Americans in the 21st century. AB - OBJECTIVES: To provide a broad overview of the role of the individual, the physical environment, and the social environment on health and functioning in older adults (65 and older), and to highlight interventions and recommendations for action on each of these levels. DATA SOURCES: Published studies and government reports on health and functioning in older Americans and on the individual, social, and physical environmental contributors to health were identified through journal and government documents review and computer library searches of medical and social science data bases for 1980-1999. STUDY SELECTION: Preference was given to published studies and government reports that focused specifically on behavioral and environmental contributors and barriers to health promotion in Americans 65 and older and/or that highlighted creative interventions with relevance to this population. Both review articles and presentations of original research were included, with the latter selected based on soundness of design and execution and/or creativity of intervention described. DATA EXTRACTION: Studies were examined and their findings organized under three major headings: (1) behavioral risk factors and risk reduction, including current government standards for prevention and screening; (2) the role of the physical environment; and (3) the role of the social environment in relation to health promotion of older adults. DATA SYNTHESIS: Although most attention has been paid to the role of behavioral factors in health promotion for older adults, a substantial body of evidence suggests that physical and social environmental factors also play a key role. Similarly, interventions that promote individual behavioral risk reduction and interventions targeting the broader social or physical environment all may contribute to health in the later years. CONCLUSIONS: With the rapid aging of America's population, increased attention must be focused on health promotion for those who are or will soon be older adults. Promising intervention strategies addressing the individual, the physical environment, and the social environment should be identified and tested, and their potential for replication explored, as we work toward a more comprehensive approach to improving the health of older Americans in the 21st century. PMID- 11067573 TI - Sources of attrition in a church-based exercise program for older African Americans. AB - Among 123 older African Americans recruited into a church-based exercise program, 43% had dropped out within four months. Compared to those who did not drop out, drop outs had lower levels of education, energy to do activities, energy to exercise, and self-ratings of health, all based on measures taken before the class. Over half of those who dropped out cited non-exercise related health problems, and 17% caregiver responsibilities. Of those who dropped out, half said they would continue to exercise and 32% said they intended to start within the next six months. PMID- 11067574 TI - Health Care Financing Administration (HCFA), the federal agency that administers the Medicare program. PMID- 11067575 TI - Electrical stimulation: a reflection on current clinical practices. AB - This paper briefly reviews the basic principles of several clinical applications of electrical stimulation for therapeutic purposes. It is intended to facilitate the integration of electrical stimulation into routine clinical practice by clarifying the terminology and standard conventions of the field, explaining the delivery capabilities of common electrical stimulators commercially available for clinical use, summarizing several examples of evidence-based therapeutic applications, and providing guidelines for selection of most commonly used treatment parameters. Rather than an exhaustive survey of the field, the presentation touches broadly on guidelines for use of transcutaneous electrical stimulation employing surface electrodes for the purposes of analgesia (TENS), drug delivery (iontophoresis), or neuromuscular rehabilitation (NMES), as well as other selected clinical applications. The paper is a general review of common clinical practices of electrotherapy and should serve as an introduction to the important factors for clinicians to consider when contemplating electrical stimulation as a treatment option. PMID- 11067576 TI - A critical review of neuromuscular electrical stimulation for treatment of motor dysfunction in hemiplegia. AB - The purpose of this review is to critically assess the clinical efficacy of neuromuscular electrical stimulation in treating motor dysfunction in hemiplegia. Three distinct applications are reviewed in the areas of motor relearning, shoulder dysfunction, and neuroprostheses. Assessment of clinical efficacy and recommendations on clinical implementation are based on the weight of published scientific evidence. With respect to motor relearning, evidence supports the use of neuromuscular electrical stimulation to facilitate recovery of muscle strength and coordination in hemiplegia. However, effects on physical disability are uncertain. With respect to shoulder dysfunction, neuromuscular electrical stimulation decreases shoulder subluxation, at least in the short term. However, effects on shoulder pain and disability are also uncertain. With respect to neuroprosthesis systems, clinically deployable upper extremity systems must await the development of more sophisticated control methods and greater fundamental understanding of motor dysfunction in hemiplegia. The evidence for clinical feasibility of lower extremity neuroprostheses is stronger, and investigations on clinical efficacy should be pursued. In summary, the application of neuromuscular electrical stimulation for motor relearning and shoulder dysfunction are ready for more rigorous scientific and clinical assessment via large, multicenter, randomized clinical trials. However, additional investigations are needed to demonstrate the clinical feasibility of neuroprostheses applications. PMID- 11067577 TI - Electrical stimulation for pressure sore prevention and wound healing. AB - This paper reviews applications of therapeutic electrical stimulation (ES) specific to wound healing and pressure sore prevention. The application of ES for wound healing has been found to increase the rate of healing by more than 50%. Furthermore, the total number of wounds healed is also increased. However, optimal delivery techniques for ES therapy have not been established to date. A study of stimulation current effects on wound healing in a pig model has shown that direct current (DC) stimulation is most effective in wound area reduction and alternating current (AC) stimulation for wound volume reduction at current densities of 127 microA/cm2 and 1,125 microA/cm2, respectively. Preliminary studies have been carried out at two research centers to assess the role of ES in pressure sore prevention. Surface stimulation studies have shown that ES can produce positive short-term changes in tissue health variables such as regional blood flow and pressure distribution. The use of an implanted stimulation system consisting of intramuscular electrodes with percutaneous leads has been found to produce additional long-term changes. Specifically, gluteal muscle thickness increased by 50% with regular long-term ES application concurrent with a 20% decrease in regional interface pressures and increased tissue oxygen levels. These findings indicate that an implantable ES system may have great potential for pressure sore prevention, particularly for individuals who lack sensation or who are physically unable to perform regular independent pressure relief. PMID- 11067578 TI - Neuroprosthetic applications of electrical stimulation. AB - Neural prostheses are a developing technology that use electrical activation of the nervous system to restore function to individuals with neurological impairment. Neural prostheses function by electrical initiation of action potentials in nerve fibers that carry the signal to an endpoint where chemical neurotransmitters are released, either to affect an end organ or another neuron. Thus, in principle, any end organ under neural control is a candidate for neural prosthetic control. Applications have included stimulation in both the sensory and motor systems and range in scope from experimental trials with single individuals to commercially available devices. Outcomes of motor system neural prostheses include restoration of hand grasp and release in quadriplegia, restoration of standing and stepping in paraplegia, restoration of bladder function (continence, micturition) following spinal cord injury, and electrophrenic respiration in high-level quadriplegia. Neural prostheses restore function and provide greater independence to individuals with disability. PMID- 11067579 TI - Reduction of costs of disability using neuroprostheses. AB - The lifetime costs associated with spinal cord injury are substantial. Assistive technology that reduces complications, increases independence, or decreases the need for attendant services can provide economic as well as medical or functional benefit. This study describes two approaches for estimating the economic consequences of implanted neuroprostheses utilizing functional electrical stimulation. Life care plan analysis was used to estimate the costs of bladder and bowel care with and without a device restoring bladder and bowel function and to compare these with the costs of implementing the device. For a neuroprosthesis restoring hand grasp, the costs of implementation were compared to the potential savings in attendant care costs that could be achieved by the use of the device. The results indicate that the costs of implementing the bladder and bowel system would be recovered in 5 years, primarily from reduced costs of supplies, medications, and procedures. The costs of the hand grasp neuroprosthesis would be recovered over the lifetime of the user if attendant time was reduced only 2 hours per day and in a shorter time if attendant care was further reduced. Neither analysis includes valuation of the quality of life, which is further enhanced by the neuroprostheses through restoration of greater independence and dignity. Our results demonstrate that implantable neuroprosthetic systems provide good health care value in addition to improved independence for the disabled individual. PMID- 11067580 TI - Functional electrical stimulation equipment: a review of marketplace availability and reimbursement. AB - Functional electrical stimulation (FES) is a rehabilitation tool that has broad application in disability management for improving consumer health and independence. This review examines the availability and delivery of electrical stimulation equipment in a managed care environment, focusing particularly on recent advances and marketplace influences. New electrical stimulation products that are unique in their ability to improve function after disease or injury over conventional drug therapy, surgical intervention, or other rehabilitation techniques are described. Research directions, including new uses for existing products to expand patient indications, are discussed. Guidelines to assist providers and developers of FES technology with managing the reimbursement process are provided. The successful introduction of recent FES products should pave the way for even more exciting developments. However, reimbursement requires careful and early planning to ensure that FES technologies are available to people who may benefit from them. PMID- 11067581 TI - Electrical stimulation: a societal perspective. AB - Societal perspective on functional electrical stimulation is colored by media influence, popular thought, and political climate as much as by the science that supports it. The purpose of this article is to examine how these influences facilitate or inhibit the application of electrical stimulation in today's world and to describe the challenges facing the use of electrical stimulation in the future. Emphasis will be placed on perceived need, cost, and available resources and how these factors must be addressed to utilize functional electrical stimulation successfully in society. PMID- 11067582 TI - Ethical issues and practical problems in preimplantation genetic diagnosis. AB - Author argues the preimplantation genetic diagnosis is ethically acceptable for currently intended uses, but a variety of ethical and practical issues will arise with further development of this technology. PMID- 11067583 TI - Cell and molecular biological challenges of ICSI: ART before science? AB - Authors discuss the possible genetic and cell biological risks to offspring conceived by ICSI in relation to the lack of fundamental research using relevant animal models. PMID- 11067584 TI - Genetic research as therapy: implications of "gene therapy" for informed consent. AB - Authors argue that characterization of gene transfer research as "gene therapy" has compromised informed consent in the current environment of regulatory exceptions, routinized consent, fostered therapeutic misconceptions, and oversold research. PMID- 11067585 TI - Commentary: reconstruing genetic research as research. AB - Author argues that although RAC has succumbed to the conflation of research and therapy as well as the problematic reduction of therapeutic criteria to compassion, its counter-cultural efforts to resist this conflation contributed to the movement to disband it. She also seeks to demonstrate how RAC has, in its work, begun to embody recommendations regarding informed consent. PMID- 11067586 TI - Killing and allowing to die: another look. AB - Author offers an analytic defense of the coherence of the distinction between "killing" and "allowing to die," based on contemporary philosophy of action and intention. PMID- 11067587 TI - Pharmaceuticals: managing managed care and recent FDA responses. PMID- 11067588 TI - The emerging technology and application of preimplantation genetic diagnosis. AB - Authors argue that although preimplantation genetic diagnosis is a promising new reproductive technology that can prevent birth defects and other devastating inherited diseases, PGD poses the risk of misdiagnosis and misuse. PMID- 11067589 TI - Legislative activity: physician-assisted suicide under managed care. PMID- 11067590 TI - Insurance: Massachusetts court denies ERISA preemption of "Any Willing Provider" Act. PMID- 11067591 TI - Insurance: Seventh Circuit enforces ERISA plan summary even without reliance by insured. PMID- 11067592 TI - Legislative activity: DOJ gives Oregon's Death with Dignity Act preliminary approval. PMID- 11067593 TI - Managed care: HMOs liable for bad faith, cost-motivated refusal to authorize care. PMID- 11067594 TI - Managed care: ERISA held not to preempt state tax on health care facilities. PMID- 11067595 TI - IVF shared-risk programs. PMID- 11067596 TI - Federal privacy legislation. PMID- 11067597 TI - Introduction: death and dying behind bars--cross-cutting themes and policy imperatives. PMID- 11067598 TI - Compassionate release from New York State prisons: why are so few getting out? AB - Author notes that the New York State compassionate release program, which permits the early release of dying inmates, has been ineffective due to both flaws in the statutory scheme and the program's implementation. He suggests changes to broaden the program without increasing risks to society. PMID- 11067599 TI - Commentary: is it politic to limit our compassion? AB - Author explores the relationship between compassion for the terminally ill in the face of "tough on crime" politics and changing epidemiology of AIDS in prison. PMID- 11067600 TI - Commentary: a personal view on palliative and hospice care in correctional facilities. AB - Author notes that the provision of palliative and hospice care in correctional settings requires a multidisciplinary team approach, involving not only health care providers, but also corrections managers and their employees. All facets of an inmate's life must be reviewed in terms of end-of-life decisions, management, and palliative care. PMID- 11067601 TI - Informed consent and the refusal of medical treatment in the correctional setting. AB - Authors note that the legal framework courts use for analyzing patients' decisions to refuse treatment does not accommodate the ethical issues arising when such decisions are made by patients in correctional settings. In lieu of the traditional model, they propose a more cautious approach when inmates refuse life sustaining medical treatment. PMID- 11067602 TI - The ethics of end-of-life care for prison inmates. AB - Author contends that the philosophical arguments involving the value of persons, social contract theory, the definition of justice, the notion of just desserts, and a utilitarian calculus of societal benefits and burdens provide support for an ethical imperative to provide end-of-life care to dying prisoners. PMID- 11067603 TI - Suicide in adult correctional facilities: key ingredients to prevention and overcoming the obstacles. AB - Author notes that inmate suicide continues to pose a serious public health problem within correctional facilities. He reviews the problem, the key components to comprehensive suicide prevention programming, and the obstacles to prevention. PMID- 11067604 TI - Death and dying in prison in Australia: national overview, 1980-1998. AB - Author discusses the role of the Australian Institute of Criminology in monitoring deaths in people in custody in Australian prisons on a national basis. She provides an overview of deaths in prison custody between 1980 and 1998, noting a shift in the cause of death of Indigenous prisoners from natural causes to self-infliction. PMID- 11067605 TI - Disability & ADA: Supreme Court clarifies the meaning of disability under ADA. PMID- 11067606 TI - Antitrust: Third Circuit rejects exception to Noerr-Pennington Doctrine creating split among circuits. PMID- 11067607 TI - Constitutional law: Fourth Circuit upholds cocaine testing of pregnant women. PMID- 11067608 TI - Disability & ADA: Supreme Court rules on institutional confinement of disabled. PMID- 11067609 TI - Employment: NLRB and renewed efforts by physicians to unionize. PMID- 11067610 TI - Fraud & abuse: two Columbia/HCA executives found guilty of fraud. PMID- 11067611 TI - Insurance: content of insurance policies not governed by the ADA. PMID- 11067612 TI - Survey of state EMS-DNR laws and protocols. AB - Author presents the results of a survey of state emergency medical services do not-resuscitate laws and protocols that have been implemented statewide and describes their structural and operational characteristics, as well as responses from key state contacts on program challenges and issues. PMID- 11067613 TI - Bioethics and the whole: pluralism, consensus, and the transmutation of bioethical methods into gold. AB - Arguing that a consensus-based method of bioethical decision making can transform ethical pluralism into an ethical whole, author examines the theory of three consensus-based models--clinical pragmatism, ethics facilitation, and mediation- and develops a practical guide to ethics facilitation that includes a hypothetical case. PMID- 11067614 TI - Commentary: a consensus about "consensus"? AB - Authors develop the notion of "consensus," which is at the heart of the "ethical consensus method" proposed by Martin, and the three approaches from which it is drawn. PMID- 11067615 TI - Fetal protection in Wisconsin's revised child abuse law: right goal, wrong remedy. AB - Authors examine Wisconsin's recent revision of its child abuse and protection laws to address substance abuse by pregnant women. The new statute enables the state to take the fetus into protective custody. Authors argue that approaching fetal protection using a child abuse model creates a series of symbolic, conceptual, and practical problems of such severity as to undermine its justifiability as a public health measure. PMID- 11067616 TI - Commentary: mal-intentioned illiteracy, willful ignorance, and fetal protection laws: is there a lexicologist in the house? AB - Author argues that fetal protection laws result from the antiabortion agenda, not from concern for child health. Such laws derive from rhetorical dissembling, not careful crafting of public health policy. PMID- 11067617 TI - Ethical and legal aspects of sperm retrieval after death or persistent vegetative state. AB - Author notes that sperm retrieval after brain death or persistent vegetative state can sometimes be ethical, provided there is explicit prior or reasonably inferred consent. Reasonably inferred consent typically has not been possible in reported cases. PMID- 11067618 TI - Commentary: legal and ethical aspects of sperm retrieval. AB - Author argues that the adequacy of "reasonably inferred consent" as a justification for the retrieval of sperm posthumously or after persistent vegetative state suffers from definitional ambiguity, faulty comparisons, and ultimately may justify practices that are not in the interest of survivors or children yet to be conceived. PMID- 11067619 TI - Commentary: posthumous harvesting of gametes--a physician's perspective. AB - Author offers a clinical perspective on problems arising from requests for postmortem sperm and oocyte retrieval. PMID- 11067620 TI - Bajakajian: new hope for escaping excessive fines under the Civil False Claims Act. AB - Author applies the U.S. Supreme Court decision in United States v. Bajakajian to argue for the reduction of fines against health care providers under the Civil False Claims Act. She compares ex ante criminal procedural protections with ex post substantive review of large civil fines. PMID- 11067621 TI - Administrative developments: DHHS issues organ allocation final rule. PMID- 11067622 TI - Alternative medicine: Ninth Circuit reverses holding on distribution of medical marijuana. PMID- 11067623 TI - Constitutional law: state court holds Arizona law on experimentation with fetuses void. PMID- 11067624 TI - Constitutional law: state partial birth abortion statutes may be constitutional. PMID- 11067625 TI - ERISA: district court limits preemption in dual option health plan. PMID- 11067626 TI - Fraud & abuse: Fifth Circuit clarifies standing of qui tam relators. PMID- 11067627 TI - Legislative developments: California enacts nurse-to-patient ratio law. PMID- 11067628 TI - Relieving pain and foreseeing death: a paradox about accountability and blame. AB - The authors look closely at the often cited Rule of Double Effect and argue that, in some cases, a physician can legitimately claim to have acted within the Rule's parameters. In other cases, however, physicians can abuse the notion of unintended consequences. The test case they offer concerns palliative care that causes foreseeable death. PMID- 11067629 TI - Commentary: double effect--intention is the solution, not the problem. AB - While the author believes that Nuccetelli and Seay have not succeeded in replacing the Rule of Double Effect with a more workable and practical moral principle, he believes they have emphasized a frequently overlooked element: that certain clear conditions must first be met before the Rule can be applied. PMID- 11067630 TI - Are ethics committee members competent to consult? AB - The authors describe research conducted to assess the skills and knowledge base of individuals who perform ethics consultations in Maryland hospitals. The findings indicate a lack of formal educational preparation on the part of those who perform consults, and a general lack of institutional support for ethics committees. PMID- 11067631 TI - Involving study populations in the review of genetic research. AB - Genetic research can present risks to all members of a study population, not just those who choose to participate in research. The authors suggest that community based reviews of research protocols can help identify and minimize such research related risks. PMID- 11067632 TI - Hands on/hands off: why health care professionals depend on families but keep them at arm's length. AB - The authors assert that a system that requires ever greater direct and indirect participation from families must change the negative presumption that families equal trouble to one that acknowledges legitimate family interests in decision making and care delivery and treats families as partners in caregiving. PMID- 11067633 TI - Commentary: what "community review" can and cannot do. AB - The author praises Sharp and Foster's differentiation of the forms of "community review," and agrees that the discussion is far from settled. He argues that rather than attempting to define "community" by various criteria, it might be more helpful to both researchers and research subjects to enable persons to create their own communities: a process of community construction, rather than reaction. PMID- 11067634 TI - Autonomy and connectedness: a re-evaluation of Georgetown and its progeny. AB - Focusing on the notable 1964 case of Application of the President and Directors of Georgetown College and subsequent cases, the author explores the connections between parents' and children's interests in ways which tend to support a parent's autonomy and hence a parent's refusal of life-sustaining treatment. PMID- 11067635 TI - Physician value neutrality: a critique. AB - Physician value neutrality is the widely-held belief that physicians must not impose their personal values on their patients. The authors argue that this concept is flawed and thus is neither possible nor should it be pursued. PMID- 11067636 TI - Commentary: value neutrality, moral integrity, and the physician. AB - The commentator agrees, in part, with Beckwith and Peppin that value neutrality is an illusion, both ethically and legally. He asserts, however, that a physician's moral autonomy does not sanction proselytizing to promote the physician's own values. PMID- 11067637 TI - Human stem cell research: NIH releases draft guidelines for comment. PMID- 11067638 TI - ERISA and RICO: new tools for HMO litigators. PMID- 11067639 TI - Legal implications of discrimination in medical practice. PMID- 11067640 TI - Wrongful death: Oklahoma Supreme Court replaces viability standard with "live birth" standard. PMID- 11067641 TI - EMTALA: OIG/HCFA Special Advisory Bulletin clarifies EMTALA, American College of Emergency Physicians criticizes it. PMID- 11067642 TI - Disability & ADA: disparate insurance coverage for physical and psychological disabilities does not violate ADA. PMID- 11067643 TI - AIDS: Mississippi Supreme Court adopts new standard for fear of exposure to AIDS. PMID- 11067644 TI - Organ transplantation: new regulations to alter distribution of organs. PMID- 11067645 TI - Internet pharmacies: regulation of a growing industry. PMID- 11067647 TI - Clinical trials and the new good clinical practice guideline in Japan. An economic perspective. AB - Japanese clinical trials have been drastically changing in response to the implementation of the International Conference on Harmonisation-Good Clinical Practice (ICH-GCP) guideline in 1997. The most important aim of the new guideline is to standardise the quality of clinical trials in the US, European Union and Japan, but it inevitably imposes substantial costs on investigators, sponsors and even patients in Japan. The study environment in Japan differs from that in the US in several ways: (i) historical lack of a formal requirement for informed consent; (ii) patients' attitudes to clinical trials in terms of expectation of positive outcomes; (iii) the implications of universal health insurance for trial participation; (iv) the historical absence of on-site monitoring by the sponsor, with the attendant effects on study quality; and (v) the lack of adequate financial and personnel support for the conduct of trials. Implementation of the new GCP guideline will improve the ethical and scientific quality of trials conducted in Japan. It may also lead to an improved relationship between medical professionals and patients if the requirement for explicit informed consent in clinical trials leads to the provision of a similar level of patient information in routine care and changes the traditional paternalistic attitude of physicians to patients. The initial response of the Japanese 'market' for clinical trials to the implementation of the ICH-GCP guideline has been clinical trial price increases and a decrease in the number of study contracts. These changes can be explained by applying a simple demand-supply scheme. Whether clinical trials undertaken in Japan become more or less attractive to the industry in the long term will depend on other factors such as international regulations on the acceptability of foreign clinical trials and the reform of domestic healthcare policies. PMID- 11067646 TI - Patient compliance with drug therapy in schizophrenia. Economic and clinical issues. AB - The effectiveness of drug treatment in clinical practice is considerably lower than the efficacy shown in controlled studies. The lower effectiveness in practice presumably leads to lower cost effectiveness of drug treatment in real life situations compared with that demonstrated by studies based on results from controlled trials. Improved cost effectiveness in routine clinical practice would be a significant advantage in the treatment of schizophrenia, one of the most costly diseases in society. The aetiology of schizophrenia is unknown, and there is no cure. The main aims of therapy with antipsychotic medication include the effective relief of symptoms without the introduction of adverse effects or serious adverse events, improved quality of life, cost effectiveness and a positive long term outcome. The older classical antipsychotic drugs do not always meet these requirements because of their well-known limitations, such as a lack of response in a subgroup of individuals with schizophrenia and intolerable adverse effects. There has long been a need for new antipsychotics that can ameliorate more symptoms and have no or few adverse effects. Some of the recently introduced antipsychotics have been shown to be more effective in certain clinical situations and to have a more favourable adverse effect profile than the classical antipsychotics. A major factor contributing to the lower effectiveness of drug treatment is noncompliance, which may be very high in schizophrenia. There are several factors influencing compliance, including drug type and formulation, patient, disease status, physician, health care system, community care and family. There have been very few studies of compliance improvement strategies in schizophrenia or, indeed, in medicine in general. Current methods are relatively complex and there are differing opinions on their effectiveness. There are several ways to increase compliance in schizophrenia--the evidence is strongest for psychoeducative methods, changing to a new drug or using a depot formulation. However, considerably more research is needed in the field of compliance strategies. PMID- 11067648 TI - Pharmacoeconomic analysis of antidepressants for major depressive disorder in the United Kingdom. AB - OBJECTIVE: To estimate the cost effectiveness of different classes of antidepressants in the UK National Health Service. DESIGN, PATIENTS AND INTERVENTIONS: The use of the serotonin (5-hydroxytryptamine; 5-HT) and noradrenaline (norepinephrine) reuptake inhibitor (SNRI) venlafaxine was compared with that of selective serotonin reuptake inhibitors (SSRIs) and tricyclic antidepressants (TCAs) in patients with major depressive disorder (MDD). A meta analysis determined the clinical success rate, and a decision tree was constructed by interviewing general practitioners and psychiatrists. Adding pharmacological and nonpharmacological treatment costs, meta-analytic rates were applied to the decision tree to calculate the expected cost and outcome for each drug. Cost effectiveness was determined using a composite measure of outcome [symptom-free days (SFD)]. MAIN OUTCOME MEASURES AND RESULTS: The meta-analysis included data from 44 studies on 4033 patients. The highest overall efficacy rate for outpatients with MDD was with venlafaxine use (73.7%), compared with 61.4% for SSRIs and 59.3% for TCAs. Treatment with venlafaxine yielded the lowest outpatient cost for a SFD (10.53 Pounds), compared with 13.23 Pounds for SSRIs and 15.52 Pounds for TCAs (1998 values). CONCLUSIONS: Using this economic model, venlafaxine appears to be a cost-effective treatment for outpatients with MDD in the UK. PMID- 11067649 TI - Differences in attitudes, knowledge and use of economic evaluations in decision making in The Netherlands. The Dutch results from the EUROMET Project. AB - OBJECTIVE: To investigate differences in attitudes, knowledge and actual use of economic evaluations in different groups of decision-makers, and to compare the results from the Netherlands with the overall European results of the European Network on Methodology and Application of Economic Evaluation Techniques (EUROMET) project. DESIGN AND SETTING: Members of the EUROMET group conducted interviews and surveys with politicians, regulators, hospital pharmacists and physicians in The Netherlands. Three approaches of investigation could be adopted: (i) a postal questionnaire survey, (ii) semi-structured interviews, and (iii) a focus-group approach. MAIN OUTCOME MEASURES AND RESULTS: In the Netherlands, decision-makers generally have a positive attitude towards economic evaluations. Nevertheless, their actual use and knowledge of economic evaluations are still limited. Hospital pharmacists and regulators are more objective than physicians and politicians, who also base their judgements on other societal values. Hospital pharmacists and regulators have a greater knowledge of economic evaluations, and they use them more often than the other groups. Most decision makers do not want to base their decisions strictly on a cost-effectiveness ranking alone. Our findings were similar to the findings in other European countries. CONCLUSIONS: Decision-makers prefer to make their own broad comparisons of advantages and disadvantages, and do not base their decisions solely on a single summary measure. PMID- 11067650 TI - Cost effectiveness of epoetin-alpha to augment preoperative autologous blood donation in elective cardiac surgery. AB - OBJECTIVE: The objective of this study was to assess the cost effectiveness of using epoetin-alpha (erythropoietin) to augment preoperative autologous donation (PAD) of blood prior to elective cardiac surgery. DESIGN AND SETTING: We designed a decision-analytic model incorporating the risk of receiving allogeneic blood, the costs of blood products, the likelihood of developing transfusion-related diseases, the costs of transfusion-related diseases and their impact on life expectancy, and the effect of epoetin-alpha on the probability of transfusion. INTERVENTIONS: The efficacy of epoetin-alpha was derived from data from a meta analysis of published randomised trials comparing the use of epoetin-alpha to augment PAD with the use of PAD alone. Estimates for the other parameters were obtained by a systematic review of the literature. MAIN OUTCOME MEASURES AND RESULTS: The use of epoetin-alpha reduced the proportion of patients receiving allogeneic transfusions by 60% (from 31.6 to 12.7%). However, this led to only a modest benefit of 0.000035 life years gained per patient and an incremental cost per life year gained of $Can44.6 million (1998 Canadian dollars). A detailed sensitivity analysis confirmed that the cost-effectiveness ratio was larger than that which is generally considered acceptable. CONCLUSIONS: Our study indicates that the use of epoetin-alpha to reduce perioperative allogeneic transfusions in cardiac surgery is not cost effective. PMID- 11067651 TI - Economic evaluations of influenza vaccination in healthy working-age adults. Employer and society perspective. AB - OBJECTIVE: To determine what benefits to the employer and to society are associated with influenza (flu) vaccination in healthy adults. DESIGN AND METHODS: We performed a literature review concerning cost-benefit and cost effectiveness evaluations of influenza vaccination in healthy, working-age adults. Up to the end of 1999, we found 6 published economic evaluations on the use of influenza vaccine in healthy, working-age adults: 3 prospective studies, 1 retrospective evaluation and 2 model-based simulations. Evaluations were performed from the perspective of an employer or society. Costs were reported in the local currency used in the published evaluation, with conversions into US dollars (when not provided in the article), for comparative purposes only, according to the exchange rates of June 8 1998. MAIN OUTCOME MEASURES AND RESULTS: Estimations of the cost-benefit of vaccination, compared with a no vaccination strategy, varied widely from a net loss of $US106.59 per infection averted in one study to savings of varying sizes in the 5 others (savings ranged from $US2.58 per dollar invested to $US46.85 per vaccinee). Studies differed in the definition of illness and the measurement of costs associated with vaccination or illness. CONCLUSIONS: Decision makers have not yet extended existing vaccine recommendations to cover healthy, working-age adults, partly because of the disparity among economic studies in their methods of estimating costs and measuring effects. However, the published studies seem to suggest that influenza vaccination in the healthy, working adult would be a cost-effective health intervention, at least from the perspective of an employer. PMID- 11067652 TI - Economic assessment of the secondary prevention of ischaemic events with lysine acetylsalicylate. AB - OBJECTIVE: To analyse the economic benefits, in comparison with placebo, of the secondary prevention of ischaemic stroke and myocardial infarction (MI) with lysine acetylsalicylate (Kardegic) in patients with a history of ischaemic stroke, MI or stable and unstable angina pectoris. DESIGN AND SETTING: This was a modelling study from the perspectives of direct medical costs, the social security system and society in France. METHODS: Efficacy data for the secondary prevention of ischaemic events were derived from the Antiplatelet Trialists' Collaboration meta-analysis on antithrombotics. The rates and costs of ischaemic disease and of serious gastrointestinal adverse affects arising from long term aspirin treatment, as well as the costs of treatment with lysine acetylsalicylate, were taken from published sources, using French data where possible. RESULTS: From the social security perspective, the estimated cost effectiveness ratios show that the prevention of MI in patients with a history of unstable angina (with a 1-year follow-up) is a cost-saving strategy, with net benefits ranging from $US5703 (1996 prices) per avoided MI for lysine acetylsalicylate 300 mg/day to $US5761 per avoided MI for lysine acetylsalicylate 75 mg/day. The prevention of MI and stroke is also a cost-saving strategy in patients with prior MI [net benefits in a 2-year follow-up (5% discount rate) ranging from $US15 to $US494 per avoided MI and from $US37 to $US1170 per avoided stroke]. This was also true in patients with prior ischaemic stroke (net benefits in a 3-year follow-up ranging from $US610 to $US2082 per avoided MI and from $US176 to $US599 per avoided stroke). Finally, a 4-year follow-up in patients with a history of stable angina pectoris shows that prophylactic treatment with lysine acetylsalicylate is associated with net costs per avoided MI, ranging from $US4375 to $US3608 per avoided event. Sensitivity analysis confirmed that prophylaxis with lysine acetylsalicylate in patients at high risk of cardiovascular and cerebrovascular events results in savings in social security expenditure. CONCLUSIONS: Our results underline the high economic benefit of using lysine acetylsalicylate to prevent secondary ischaemic stroke and MI in patients at high risk of cardiovascular and/or cerebrovascular events, leading to savings for the social security system and society. PMID- 11067654 TI - Measuring public health performance: a call to action. PMID- 11067653 TI - Antiviral therapies for herpes zoster infections. Are they economically justifiable? AB - Antiviral treatment of herpes zoster is controversial because of uncertain benefits and relatively high costs. Most studies show that antiviral therapy lessens acute herpes zoster symptoms and postherpetic neuralgia (PHN). Current clinical recommendations support antiviral treatment of severely symptomatic herpes zoster in all adults, and mild herpes zoster in those 50 or 60 years of age or older. However, it is unclear if these recommended strategies are cost effective. Published studies of herpes zoster costs and the effect of antiviral therapy on costs and quality of life have significant variation in study design and results, as well as many shortcomings in the data. Thus, definitive economic recommendations cannot be made based on the present data. Another approach, which we have used, is to develop a 'reference case' analysis using decision-analysis techniques and the available data to estimate the incremental cost effectiveness of antiviral treatment in patients of differing age and herpes zoster severity. In the baseline analysis, parameter values and assumptions were consistently slightly biased against antiviral use. Effectiveness was measured in quality adjusted life years (QALYs). We assumed that antiviral treatment did not change PHN risk, but decreased PHN duration in patients older than 50 years. PHN risk increased with age and with acute herpes zoster severity as seen in published data. Mild acute herpes zoster was assumed to have a utility value of 0.9 and severe acute herpes zoster a value of 0.7 on a scale where 0 = death and 1 = perfect health. Treating mildly symptomatic acute herpes zoster cost $US89,200/QALY gained in 40-year-olds, $US47,700/QALY in 60-year-olds and $US40,700/QALY in 70-year-olds (1995 values). Results were most sensitive to variation of antiviral costs (baseline $US134), but changes in acute symptom relief, PHN risk, duration, costs and utility, and antiviral effect on PHN duration increased costs/QALY above $US50,000 in 60- and 70-year-olds in extremes of parameter ranges. However, no variation resulted in treatment of mild illness in 40-year-olds to fall below $US50,000/QALY gained. Treatment of severe acute herpes zoster cost $US29,700, $US18,000 and $US16,500/QALY gained in 40-, 60- and 70-year-olds, respectively. Results were sensitive to variation of antiviral costs (> $US225) and acute symptom relief (< 21%) in 40-year-olds. Based on this analysis, antiviral therapy of herpes zoster seems economically justifiable for mildly symptomatic acute herpes zoster in patients aged 50 years and older, and for severely symptomatic acute herpes zoster in all adults. PMID- 11067655 TI - Performance measurement and performance standards: old wine in new bottles. PMID- 11067656 TI - Using the essential services as a foundation for performance measurement and assessment of local public health systems. AB - Efforts are under way to develop a performance measurement monitoring system for state and local public health systems and to develop a strategic planning tool for local public health systems. The development of these measures is being based on the Essential Public Health Services. This article provides the rationale for why the Essential Services offer a good framework for identifying, analyzing, and evaluating public health activities. The article also reviews the history of local public health and the development and application of the Essential Public Health Services and their predecessor frameworks such as the core functions, the organizational practices, and the essential elements. PMID- 11067657 TI - Can public health performance standards improve the quality of public health practice? AB - Recent developments suggest that a national public health performance standards program could succeed in improving the quality of public health practice. Public health standards also may be useful for enhancing accountability and strengthening the science base of public health practice. For national public health performance standards to have a substantial influence on the quality of public health practice, several important issues must be addressed. These include agreement as to the ultimate purpose and appropriate unit of measurement, delineation of the specific qualities to be measured, and expansion of strategies to promote widespread use of public health practice standards. PMID- 11067658 TI - The role of states in ensuring essential public health services: development of state-level performance measures. AB - The Association of State and Territorial Health Officials (ASTHO) has worked with other public health partners across the country to develop National Public Health Performance Standards, nationally recognized measures by which state public health systems can compare themselves with similar systems across the country. The lead federal agency is the Centers for Disease Control and Prevention (CDC), and other partners include the Public Health Foundation, the American Public Health Association, the National Association of City and County Health Officials, and the National Association of Local Boards of Health. Both challenges and opportunities emerged during the development of the state public health system standards. PMID- 11067659 TI - Profiles in courage: evolution of Florida's quality improvement and performance measurement system. AB - As public health begins to embrace performance standards, it is important to critically examine state and local health department quality improvement and performance measurement systems. In this article, Florida's decade-long evolution of quality improvement and performance measurement as well as their integration are described. Lessons for other states are discussed along with implications for implementing performance standards for state and local health departments. PMID- 11067660 TI - Local health department capacity and performance in New Jersey. AB - The Institute of Medicine's report in 1988 has spawned numerous efforts to strengthen the nation's public health system. Performance monitoring and public health practice standards have received considerable attention. All local health departments were surveyed in New Jersey to assess performance of core functions, 10 organizational practices, and organizational capacity in terms of staffing, communications, and computer capabilities. Overall, core function performance was measured at 59.7 percent and four organizational practices were measured at less than 50 percent performed. Information from this study will help direct efforts to strengthen New Jersey's public health system. PMID- 11067661 TI - Building the science base for public health practice. AB - This article explores the need for and current state of the science base in public health practice. In addition, it discusses how the National Public Health Performance Standards Program will help build the science base in the future and how this can have a positive effect on public health practice and community health status. PMID- 11067662 TI - Conceptual and methodological issues in public health performance measurement: results from a computer-assisted expert panel process. AB - As part of the developmental process for the National Public Health Performance Standards Program, the CDC convened a group of experts in the fields of public health practice and research to evaluate key conceptual and methodological issues involved in measuring the performance of public health organizations. Participants engaged in a nominal group process and an electronic polling exercise designed to elicit expert opinions about these issues. Results revealed broad consensus around the need for measurement systems that support quality improvement and accountability applications, with scientific investigation viewed as an important but secondary objective of measurement. Substantial variation was observed in perceptions about the importance of specific measurement concepts and methods. Results highlight the need for performance measurement systems to reflect multiple organizational perspectives in their design and implementation. PMID- 11067663 TI - Performance measurement in public health: conceptual and methodological issues in building the science base. AB - The use of scientific methods for examining performance in the field of public health has lagged behind comparable efforts in medical practice. Accomplishments in other health care settings demonstrate that performance measurement systems can, if pursued rigorously and systematically, advance scientific knowledge and enhance the production of information to support improvements in public health practice. Numerous conceptual and methodological issues need to be addressed in order to use public health performance measurement processes for scientific inquiry. Nonetheless, the reward for careful analytic work in this area will be an expanding body of evidence to inform policy and administrative decision making in public health. PMID- 11067664 TI - How can performance standards enhance accountability for public health? AB - This article focuses on how a national system of measuring public health performance can help enhance accountability for public health. It describes the trend toward increased accountability in public health; provides an overview of the issues and challenges public health practitioners face in demonstrating how the resources they spend and programs they operate contribute to improved community health status and suggests how the results of participating in the National Public Health Performance Standards Program can help. PMID- 11067665 TI - How can performance standards strengthen accountability for public health? AB - Performance standards with both process and outcome measures can lead to greatly increased accountability for public health and a major leadership position in U.S. health care. Accreditation of health departments should become part of the accountability process. PMID- 11067666 TI - The National Public Health Performance Standards Program: will it strengthen governance of local public health? AB - Governing bodies such as local boards of health are the government authority ultimately accountable for public health at the local level. The National Public Health Performance Standards Program (NPHPSP) provides governing bodies guidance in their oversight of the public health system and the provision of essential public health services. Using the NPHPSP's standard guidelines and criteria, local public health governing boards can identify assets and needs for public health improvements, coordinate existing services and programs, and target their advocacy for public health resources. The NPHPSP provides governing boards a means for ensuring that local public health systems coordinate their efforts to improve accountability, quality, and evidence for community health. PMID- 11067667 TI - The quest for an accurate accounting of public health expenditures. AB - This article describes one effort to develop management tools that will help public health administrators and policy makers implement comprehensive public health strategies. It recounts the ongoing development of a methodology through which the Essential Public Health Services can be related to public health budgets, appropriations, and expenditures. Through three pilot projects involving: (1) nine state health agencies, (2) three local health agencies, and (3) all local jurisdictions and the state health agency in one state, a workable methodology for identifying public expenditures for comprehensive public health programming has been identified. PMID- 11067668 TI - Outpatient commitment in mental health: is coercion the price of community services? PMID- 11067669 TI - Percutaneous vertebroplasty. PMID- 11067670 TI - Oncology. PMID- 11067671 TI - The arts and humanities in health care and education. A statement from the Work Group on the Arts and Humanities of the International Work Group on Death, Dying and Bereavement. PMID- 11067672 TI - Advance directives: legal remedies and psychosocial interventions. AB - Although the legal basis to refuse life supports is firmly embedded in the laws of all 50 states, there is evidence that a gap exists between patients' preferences and physicians' actions. Patients and their families have increasingly begun to turn to the courts for redress, requesting damages when a physician has ignored their request to forgo life-sustaining treatment. This article explores the reasons why patients' end-of-life medical choices are often ignored by the medical profession and the results of recent attempts to remedy these situations through the courts. Implications for practice are discussed, including practical suggestions for increasing the likelihood that a patient's wishes will be respected by medical providers. PMID- 11067673 TI - Hastening death: a comparison of two end-of-life decisions. AB - This study determined the relationship of psychosocial and background variables to elders' end-of-life (EOL) decision preferences. Responding to 5 EOL decision scenarios depicting terminally ill elders, 200 elders aged 60-90 indicated preferences regarding extending life (EL), refusing treatment (RT), and assisted suicide (AS). They were also assessed on religiosity, values, fear of death, locus of control, health, socioeconomic status, and age. Results of multinomial logistic regression indicated that EOL decisions of three groups (favoring EL, favoring RT, and favoring both AS and RT) were significantly influenced by religiosity, value for preservation of life, value for quality of life, fear of death, and locus of control belief. The importance of safeguarding older adults' autonomy in EOL decisions was stressed. PMID- 11067674 TI - Using study circles in the workplace as an educational method of facilitating readjustment after a traumatic life experience. AB - While employees who have undergone a traumatic life experience (TLE) are often referred for counseling and therapy, most organizations do not prepare managers and co-workers for the unique issues posed by TLE employees when they first return to the workplace. The Study Circle, a "democratic," interactive adult learning format, is introduced to make up for this lack by educating managers and co-workers about the various stages in the trauma recovery process and teaching them how to facilitate a TLE employee's re-entry to the workplace. PMID- 11067675 TI - Medicare program; Medicare+Choice program. Health Care Financing Administration (HCFA), HHS. Final rule with comment period. AB - This final rule with comment period responds to comments on the June 26, 1998 interim final rule that implemented the Medicare+Choice (M+C) program and makes revisions to those regulations where warranted. We also are making revisions to the regulations that are necessary to reflect the changes to the M+C program resulting from the Balanced Budget Refinement Act of 1999 (BBRA). Revisions to the regulations reflecting changes in the law made by the BBRA are subject to public comment. Issues discussed in this rule include eligibility, election, and enrollment policies; marketing requirements; access requirements; service area and benefit policy; quality improvement standards; payment rates, risk adjustment methodology, and encounter data submission; provider participation rules; beneficiary appeals and grievances; contractual requirements; and preemption of State law by Federal law. This final rule also addresses comments on the interim final rule published on December 2, 1997, which implemented user fees for section 1876 risk contractors for 1998, and formed the basis for the M+C user fee provisions in the June 26, 1998 interim final rule, and the provider-sponsored organization (PSO) interim final rule published April 14, 1998. PMID- 11067676 TI - Standards of compliance for abortion-related services in family planning services projects. Office of Population Affairs, OPHS, DHHS. Final rules. AB - The rules issues below revise the regulations that apply to grantees under the federal family planning program by readopting the regulations, with one revision, that applied to the program prior to February 2, 1988. Several technical changes to the regulation are also made to remove and/or update obsolete regulatory references. The effect of the revisions made by the rules below is to revoke the compliance standards, promulgated in 1988 and popularly known as the "Gag Rule," that restricted family planning grantees from providing abortion-related information in their grant-funded projects. PMID- 11067677 TI - Medicare program; prospective payment system for home health agencies. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This final rule establishes requirements for the new prospective payment system for home health agencies as required by section 4603 of the Balanced Budget Act of 1997, as amended by section 5101 of the Omnibus Consolidated and Emergency Supplemental Appropriations Act for Fiscal Year 1999 and by sections 302, 305, and 306 of the Medicare, Medicaid, and SCHIP Balanced Budget Refinement Act of 1999. The requirements include the implementation of a prospective payment system for home health agencies, consolidated billing requirements, and a number of other related changes. The prospective payment system described in this rule replaces the retrospective reasonable-cost-based system currently used by Medicare for the payment of home health services under Part A and Part B. PMID- 11067678 TI - Medical devices; effective date of requirement for premarket approval for a class III preamendments obstetrical and gynecological device. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule to require the filing of a premarket approval application (PMA) or a notice of completion of product development protocol (PDP) for a Group 1 preamendments class III device, the obstetric data analyzer intended to analyze data from fetal and maternal monitors during labor and to warn of possible fetal distress. The agency has summarized its findings regarding the degree of risk of illness or injury designed to be eliminated or reduced by requiring the device to meet the statute's approval requirements and the benefits to the public from the use of the devices. PMID- 11067679 TI - Determining disability and blindness; substantial gainful activity guides. Social Security Administration. Final rules. AB - We are revising our rules to reflect amendments to the Social Security Act (the Act) concerning the trial work period and the disability insurance reentitlement period. We are also clarifying certain standards we use to determine whether work is substantial gainful activity and whether an individual is entitled to a trial work period, thereby further explaining how we determine disability under titles II and XVI of the Act. PMID- 11067680 TI - Chemotherapy sensitivity and resistance assays. PMID- 11067681 TI - Magnetic resonance imaging of the breast: differential diagnosis of a breast lesion to avoid biopsy. PMID- 11067682 TI - Magnetic stimulation in the treatment of urinary incontinence in adults. PMID- 11067683 TI - Sacral nerve stimulation for the treatment of urinary urgency/frequency. PMID- 11067684 TI - Salvage high-dose chemotherapy with allogeneic stem-cell support for relapse or incomplete remission following high-dose chemotherapy with autologous stem-cell transplantation for hematologic malignancies. PMID- 11067685 TI - Special report: relative effectiveness of treatment programs and components of treatment for anorexia nervosa. AB - PLEASE NOTE: This draft Special Report is an evidence-based analysis of the relative effectiveness of different treatment programs and components of treatment for anorexia nervosa. It does not attempt to address the question as to whether the TEC criteria are met. PMID- 11067686 TI - Recent developments in health insurance, life insurance, and disability insurance case law. PMID- 11067687 TI - Recent developments in insurance coverage litigation. PMID- 11067688 TI - Recent developments in medicine and law. PMID- 11067689 TI - Recent developments in products, general liability, and consumer law. PMID- 11067690 TI - Recent developments in public regulation of insurance. PMID- 11067691 TI - The Veterans Millennium Health Care and Benefits Act. Department of Veterans Affairs. Final rule. AB - This document amends Department of Veterans Affairs (VA) adjudication regulations to reflect changes made by the Veterans Millennium Health Care and Benefits Act. These changes concern payment of dependency and indemnity compensation to the surviving spouses of certain former prisoners of war; the provision of health care, education and home loan benefits to surviving spouses upon termination of their remarriages; and the addition of bronchiolo-alveolar carcinoma to the list of diseases that VA presumes are the result of exposure to radiation during active military service. PMID- 11067692 TI - Wireless medical telemetry service. Federal Communications Commission. Final rule. AB - This document allocates new spectrum and establishes rules for a Wireless Medical Telemetry Service (WMTS) that allows potentially life-critical equipment to operate on an interference-protected basis. Medical telemetry equipment is used in hospitals and health care facilities to transmit patient measurement data, such as pulse and respiration rates to a nearby receiver, permitting greater patient mobility and increased comfort. This action will increase the reliability of medical telemetry equipment. PMID- 11067693 TI - Who knows your medical secrets? It's getting easier for health marketers, your boss, and your mortgage company to find out what ails you. PMID- 11067694 TI - Age-proofing your brain: can mental gymnastics and other steps help? PMID- 11067695 TI - Is an HMO for you? PMID- 11067696 TI - Safe in the sun: how to enjoy the rays without getting burned. PMID- 11067697 TI - The cyber file cabinet: dot-coms try to shift storage of medical records online. PMID- 11067698 TI - TRICARE: Civilian Health and Medical Program of the Uniformed Services (CHAMPUS); nonavailability statement requirement for maternity care. Office of the Secretary, DoD. Final Rule. AB - This final rule implements Section 712(c) of the National Defense Authorization Act for Fiscal Year 2000 (Pub. L. No. 106-65), which requires that a nonavailability-of-health-care statement shall be required for a beneficiary not enrolled in TRICARE Prime for TRICARE cost-share of maternity care services related to outpatient prenatal, outpatient or inpatient delivery, and outpatient post-partum care subsequent to the visit which confirms the pregnancy. The Act reestablishes a requirement which was previously eliminated under the broad direction of the National Defense Authorization Act for FY 1997, section 734, which removed authority for nonavailability statements (NASs) for outpatient services. Therefore, the Act changes the provisions which require an NAS for inpatient delivery, but do not require an NAS for outpatient prenatal and post partum care. The change will significantly contribute to continuity of care for maternity patients. In furtherance of that principle, and consistent with the previous policy, an NAS for maternity care shall not be required when a beneficiary has other health insurance for primary coverage. PMID- 11067699 TI - Outcomes of a small group educational intervention for urinary incontinence: health-related quality of life. AB - OBJECTIVES: This article examines change in general and condition-specific measures of health-related quality of life (HRQL) among participants in a randomized trial of a community-based intervention for urinary incontinence (UI). METHODS: Participants were randomized into intervention or wait control conditions. Participants were women aged 65 or older with urinary incontinence residing in Oklahoma. General HRQL measures included the Physical Function, Mental Health, Vitality, and Health Perceptions subscales of the Medical Outcomes Study Short Form-36. Condition-specific measures included the Impact of UI and self-management strategies. RESULTS: There were no significant group effects for the general HRQL measures. Intervention participants reported decreased Impact of UI and greater change in self-management strategies than control participants. DISCUSSION: The intervention affected condition-specific quality of life and self management but not general HRQL. The intervention's impact on quality of life involves change in how the condition is seen as impacting on life and on selection of self-management behaviors. PMID- 11067700 TI - Stage of life course and social support as a mediator of mood state among persons with disability. AB - OBJECTIVE: This research seeks to determine which aspects of social support are most effective in mediating mood state among working-age and elderly adults with disability (N = 442). METHODS: Participants were identified through random-digit dialing of telephone exchanges and administration of a disability screen. Multiple regression was used to model multiple aspects of social support while holding sociodemographic and disability indicators constant. RESULTS: Analyses revealed that network size and confidence in the reliability of helping networks are significantly and negatively related to depressed mood. Confidant support was related to lower levels of depressed mood for younger respondents only. Neither marital status, advisor support, nor social integration were related to mood. DISCUSSION: Both instrumental and emotional support are key in mediating depressed mood among this population. We conclude that all types of social support are not equally effective in mediating mood among people with disability. PMID- 11067701 TI - Differences in functional status of Hispanic versus non-Hispanic White elders: data from the Medical Expenditure Panel Survey. AB - OBJECTIVES: Few national studies have focused specifically on the functional status of Hispanic elders. We examined the prevalence of functional limitations and disabilities among Hispanic and Black elders compared to non-Hispanic Whites. METHODS: We analyzed seven measures of functional limitations, disabilities, and dependencies. Logistic regression was used to examine racial and ethnic group differences adjusting for age, gender, and education. RESULTS: Compared to non Hispanic Whites, Hispanics tended to report greater instrumental activities of daily living (IADL) dependencies and cognitive disabilities. Blacks were more likely to have activities of daily living (ADL) and IADL dependencies and require use of assistive devices compared to non-Hispanic Whites. Further adjustment for respondent status reduced differences between groups, but these models may overadjust for functional status differences. DISCUSSION: Given the projected growth of minority elders, policymakers and planners will need to consider race and ethnic differentials in functional status in determining future medical and social service needs. PMID- 11067702 TI - When it is more than a job: close relationships between home health aides and older clients. AB - OBJECTIVES: A qualitative study was conducted to investigate the types of relationships that were formed between older clients and their home health aides and to identify structural characteristics and interactive processes that facilitated various types of relationships. METHODS: Using semistructured interviews, members of 16 families and their home health aides described the relationships that developed between clients and aides and the conditions and contexts that facilitated or inhibited close relationship development. Data were coded and analyzed using a multistage process. RESULTS: Most relationships between aides and older clients were described as friendship or like one of the family, with friendship occurring most often. Several structural conditions and numerous interactive friendship processes were identified. The cognitive process of boundary setting discriminated between friendships and family-like relationships. DISCUSSION: Results show support for socioemotional selectivity theory and highlight the benefits of close relationships for older homebound adults and their home health aides. PMID- 11067703 TI - Incorporating assistive devices into community-based long-term care: an analysis of the potential for substitution and supplementation. AB - OBJECTIVES: This article examines the use of assistive devices as a part of the long-term care arrangements of community-dwelling older Americans. It examines the potential for assistive devices to substitute for and supplement personal care assistance. METHODS: Data from the Phase 2 Disability Supplements to the 1994-1995 National Health Interview Surveys are used to compare the use of personal care and equipment among persons reporting difficulty with a given activity of daily living. RESULTS: The capacity of equipment to substitute for or supplement personal care is highly task-specific and depends on the characteristics of the devices and the personal care providers. In general, those using simple devices are less likely to use informal care, whereas those using complex devices are more likely to use formal care services. DISCUSSION: Technology has the potential to confer quality of life enhancements for older persons and their caregivers and cost savings for payers. PMID- 11067704 TI - Hitting a moving target: income-related health insurance subsidies for the uninsured. AB - New government health insurance programs are likely to emphasize voluntary purchases in a market setting, with subsidies targeted at low-income populations and stress on managed care. Such programs are best structured with a guaranteed enrollment period that is as long as six months to a year. However, given that incomes change over time, errors will be made in awarding income-related subsidies for that long. These errors are assessed in simulations undertaken with longitudinal data from the Survey of Income and Program Participation. Two allocations of the subsidies, based on current income at the beginning of the enrollment period and on actual income assessed at the end, are compared for a variety of program designs. Prospective determination of subsidies is somewhat biased toward overpayment. Net overpayments amount to 5-10 percent of subsidy costs. However, prospective payment encourages participation in the subsidy program. The simulated participation rate for true eligibles is as high as 73 percent with prospective subsidies, compared to 69 percent with retrospective reconciliation. Net overpayments are slightly reduced by testing income less frequently and over longer periods. PMID- 11067705 TI - Cloning: a Jewish law perspective with a comparative study of other Abrahamic traditions. PMID- 11067706 TI - Medicare program; prospective payment system and consolidated billing for skilled nursing facilities--update. Health Care Financing Administration (HCFA), HHS. Final rule. AB - This final rule sets forth updates to the payment rates used under the prospective payment system (PPS) for skilled nursing facilities (SNFs), for fiscal year 2001. Annual updates to the PPS rates are required by section 1888(e) of the Social Security Act, as amended by the Medicare, Medicaid and State Child Health Insurance Program Balanced Budget Refinement Act of 1999, related to Medicare payments and consolidated billing for SNFs. In addition, this rule sets forth certain conforming revisions to the regulations that are necessary in order to implement amendments made to the Act by section 103 of the Medicare, Medicaid and State Child Health Insurance Program Balanced Budget Refinement Act of 1999. PMID- 11067707 TI - Cold, cough, allergy, bronchodilator, and antiasthmatic drug products for over the-counter human use; amendment of final monograph for OTC antitussive drug products. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule amending the final monograph for over-the-counter (OTC) antitussive drug products (products that relieve cough). Use of topical/inhalant products containing camphor or menthol near a flame, in hot water, or in a microwave oven may cause the products to splatter and cause serious burns to the user. As part of its ongoing review of OTC drug products, FDA is adding warnings and directions to inform consumers about these improper uses and is amending its final regulations for OTC drug labeling requirements to add this new flammability warning for antitussive drug products containing camphor or menthol. PMID- 11067708 TI - Medicare program; provisions of the Balanced Budget Refinement Act of 1999; hospital inpatient payments and rates and costs of graduate medical education. Health Care Financing Administration (HCFA), HHS. Interim final rule with comment period. AB - This interim final rule with comment period implements, or conforms the regulations to, certain statutory provisions relating to Medicare payments to hospitals for inpatient services that are contained in the Medicare, Medicaid, and State Children's Health Insurance Program Balanced Budget Refinement Act of 1999 (Public Law 106-113). These provisions relate to reclassification of hospitals from urban to rural status, reclassification of certain hospitals for purposes of payment during Federal fiscal year 2000, critical access hospitals, payments to hospitals excluded from the hospital inpatient prospective payment system, and payments for indirect and direct graduate medical education costs. Many of the provisions of Public Law 106-113 modify changes to the Social Security Act made by the Balanced Budget Act of 1997 (P.L. 105-33). These provisions are already in effect in accordance with Public Law 106-113. PMID- 11067709 TI - Medicare program; changes to the hospital inpatient prospective payment systems and fiscal year 2001 rates. Health Care Financing Administration (HCFA), HHS. Final rule. AB - We are revising the Medicare hospital inpatient prospective payment system for operating costs to: implement applicable statutory requirements, including a number of provisions of the Medicare, Medicaid, and State Children's Health Insurance Program Balanced Budget Refinement Act of 1999 (Pub. L. 106-113); and implement changes arising from our continuing experience with the system. In addition, in the Addendum to this final rule, we describe changes to the amounts and factors used to determine the rates for Medicare hospital inpatient services for operating costs and capital-related costs. These changes apply to discharges occurring on or after October 1, 2000. We also set forth rate-of-increase limits and make changes to our policy for hospitals and hospital units excluded from the prospective payment systems. We are making changes to the policies governing payments to hospitals for the direct costs of graduate medical education, sole community hospitals and critical access hospitals. We are adding a new condition of participation on organ, tissue, and eye procurement for critical access hospitals that parallels the condition of participation that we previously published for all other Medicare-participating hospitals. Lastly, we are finalizing a January 20, 2000 interim final rule with comment period (65 FR 3136) that sets forth the criteria to be used in calculating the Medicare disproportionate share adjustment in reference to Medicaid expansion waiver patient days under section 1115 of the Social Security Act. PMID- 11067710 TI - Medical use of byproduct material; policy statement, revision. Nuclear Regulatory Commission. Final policy statement; revision. AB - The Nuclear Regulatory Commission (NRC) is revising its 1979 policy statement on the medical use of byproduct material. These revisions are one component of the Commission's overall program for revising its regulatory framework for medical use, including its regulations that govern the medical use of byproduct material. The overall goals of this program are to focus NRC regulation of medical use on those medical procedures that pose the highest risk and to structure its regulations to be risk-informed and more performance-based, consistent with NRC's "Strategic Plan for Fiscal Year 1997-Fiscal Year 2002." The policy informs NRC licensees, other Federal and State agencies, and the public of the Commission's general intentions in regulating the medical use of byproduct material. PMID- 11067711 TI - Medicare program; prospective payment system for hospital outpatient services: revisions to criteria to define new or innovative medical devices, drugs, and biologicals eligible for pass-through payments and corrections to the criteria for the grandfather provision for certain Federally Qualified Health Centers. Health Care Financing Administration (HCFA), HHS. Interim final rule with comment period. AB - This interim final rule with comment period changes one criterion and postpones the effective date for two other criteria that a new device, drug, or biological must meet in order for its cost to be considered "not significant" for purposes of determining its eligibility for transitional pass-through payments. It also changes the transitional pass-through payment policy to include new single use medical devices that come in contact with human tissue and that are surgically implanted or inserted in a patient whether or not the devices remain with the patient after the patient is released from the hospital outpatient department. These policies represent a departure from those presented in the April 7, 2000 Federal Register final rule with comment period entitled, "Prospective Payment System for Hospital Outpatient Services." This interim final rule with comment period also corrects a trigger date for grandfathering of provider-based Federally Qualified Health Centers (FQHCs) to conform with the intent not to disrupt existing FQHCs with longstanding provider-based treatment that we discussed in the April 2000 final rule. Under the criteria in the April 2000 final rule with comment period, FQHCs are treated as departments of a provider without regard to the criteria for provider-based status in that document if they meet other criteria and were designated as FQHCs before 1995. Under this correction, facilities that meet those other criteria and were designated as FQHCs or "look-alikes" on or before April 7, 2000 would continue to be treated as provider-based. In addition, we are clarifying how the requirement for prior notice to beneficiaries is to be applied in emergency situations. Also, we are clarifying the protocols for off-campus departments in emergency situations. PMID- 11067712 TI - Vagus nerve stimulation for epilepsy. PMID- 11067713 TI - Endocrinology, diabetes, and metabolism. PMID- 11067714 TI - Gastroenterology and urology devices; reclassification of the extracorporeal shock wave lithotripter. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule to reclassify from class III to class II the extracorporeal shock wave lithotripter, when intended for use to fragment kidney and ureteral calculi. FDA is taking this action on its own initiative in order to assure that these devices are regulated according to the appropriate degree of regulatory control needed to provide reasonable assurance of their safety and effectiveness. PMID- 11067715 TI - Civilian Health and Medical Program of the Uniformed Services (CHAMPUS); enhancement of dental benefits under the TRICARE retiree dental program. Office of the Secretary, DoD. Interim final rule with request for comments. AB - This interim final rule implements section 704 of the National Defense Authorization Act for Fiscal Year 2000, to allow additional benefits under the retiree dental insurance plan for Uniformed Services retirees and their family members that may be comparable to those under the Dependents Dental Program. The Department is publishing this rule as an interim final rule in order to comply timely with the desire of Congress to meet the needs of retirees for additional dental coverage. Public comments are invited and will be considered for possible revisions to this rule at the time of publication of the final rule. PMID- 11067716 TI - Ceftriaxone and otitis in children: new indication. Only in special circumstances. AB - (1) Ceftriaxone, a third-generation cephalosporin antibiotic, is now licensed in France for (intramuscular) treatment of acute otitis media in children, both as first-line therapy in children under 30 months, and after failure of a first antibiotic regimen. (2) The clinical file on first-line ceftriaxone treatment is relatively bulky. In contrast, only two non comparative trials of ceftriaxone after failure of initial treatment are available. (3) According to trials of first-line treatment, the efficacy of a single intramuscular dose of ceftriaxone is equivalent to that of the sulfamethoxazole + trimethoprim combination and the amoxicillin + clavulanic acid combination, and similar to that of amoxicillin (when all these reference antibiotics are given orally for 10 days). (4) Pain at the injection site is the main adverse effect of ceftriaxone, despite the local anaesthetic (lidocaine) contained in the solvent. PMID- 11067717 TI - Idarubicin oral: new formulation. No proven benefit over 60. AB - (1) Oral idarubicin is marketed in France for the treatment of acute myeloblastic leukaemia in patients over 60 who are ineligible for intensive intravenous treatment. (2) The clinical file on oral idarubicin is limited. (3) In non comparative trials the median survival time after starting treatment with oral idarubicin, alone or combined with other cytotoxic agents, did not exceed 12 months. (4) The only comparative trial of single-agent idarubicin in this indication gave unfavourable results. Another comparative trial tested a combination comprising idarubicin, but the role of idarubicin alone cannot be determined from the results. (5) In the palliative care setting the clinical efficacy of oral idarubicin has not yet been demonstrated. (6) The main adverse events observed during oral idarubicin therapy are neutropenia, thrombocytopenia, nausea and vomiting, diarrhoea and hair loss. Their impact on patients' quality of life has not been adequately assessed. PMID- 11067718 TI - Pneumococcal vaccination for elderly subjects: license extension. Still no proof of clinical efficacy. AB - (1) In France the licensed indications of a 23-valent polyoside pneumococcal vaccine called Pneumo 23 degrees have been extended to cover people over 65. (2) The only relevant published double-blind trial showed no reduction in the risk of pneumonia or death after vaccination in this age group. Similarly, a comparative unblinded trial involving more than 26,000 elderly people showed no reduction in the risk of pneumonia. (3) Some retrospective studies suggest that the vaccine may reduce the risk of infection with bacteraemia or meningitis, with no clear protection against the serotypes usually associated with diminished susceptibility to penicillin. (4) Randomised trials have also failed to show that older pneumococcal vaccines reduce the risk of pneumonia or death. (5) The adverse effects of Pneumo 23 degrees are generally mild and local. They occur most frequently after booster injections. PMID- 11067719 TI - Interferon beta-1b and secondary progressive multiple sclerosis: licence extension. Useful, but further assessment required. AB - (1) Interferon beta-1b is now licensed to treat patients with secondary progressive multiple sclerosis. (2) The clinical file that we compiled on this indication, includes a double-blind placebo-controlled trial in which 718 patients were treated for 2-3 years. The trial is methodologically sound. (3) In this trial, interferon beta-1b, at the only dose tested (8 MIU every two days by the subcutaneous route), significantly reduced the progression of the disability linked to the disease. After 2-3 years of treatment the percentage of patients confined to a wheelchair was, in absolute values, 16.7% in the interferon beta-1b group and 24.6% in the placebo group. (4) In this trial the adverse effects linked to interferon beta-1b were already known, i.e. mainly a 'flu-like syndrome at the outset of treatment, and reactions at the injection site. (5) More than a quarter of patients on interferon beta-1b had neutralising antibodies against interferon beta-1b. More follow-up is needed to determine the possible impact of these antibodies on treatment efficacy and on the possible risk of autoimmune diseases. (6) Treatment with interferon beta-1b is still costly. PMID- 11067720 TI - Sertraline and obsessive compulsive disorder: new indication. Limited assessment. AB - (1) Sertraline, a selective serotonin reuptake inhibitor (SSRI), is now licensed in France for the treatment of obsessive-compulsive disorder in adults. (2) In this indication the clinical file is of acceptable methodological quality, but it is incomplete: sertraline has not been compared with the other two serotonin reuptake inhibitors approved in obsessive-compulsive disorder, namely fluoxetine and paroxetine. (3) Three placebo-controlled trials have demonstrated the efficacy of sertraline in obsessive-compulsive disorder. (4) In a trial versus clomipramine, sertraline was no more effective in patients able to tolerate the drug, but the rate of treatment withdrawals for adverse events was higher on clomipramine. PMID- 11067721 TI - Cutaneous reactions to topical NSAIDs. AB - (1) Cutaneous application of nonsteroidal antiinflammatory drugs (NSAIDs) can cause potentially severe skin lesions that may not be restricted to the site of application. Sunlight can be an aggravating factor. (2) Various topical NSAIDs have been implicated. The most frequently mentioned is ketoprofen. PMID- 11067722 TI - Peripheral neuropathy due to statins. AB - (1) A rare but potentially incapacitating adverse effect. PMID- 11067723 TI - Dangers of low-dose colchicine in patients with renal failure. AB - (1) Life-threatening colchicine intoxication can occur in patients with renal failure, even with low-dose regimens. (2) Diarrhoea, nausea and vomiting are the first manifestations of overdose. Colchicine must be withdrawn immediately to avoid severe complications (especially haematological and neuromuscular). (3) Colchicine must not be prescribed without first assessing creatinine clearance (using the Cockroft formula, for example), especially in elderly patients. PMID- 11067724 TI - Severe hepatic and cutaneous adverse effects with nevirapine. AB - (1) Introduce nevirapine at half the target dose, and monitor the skin, mucous membranes and liver to avoid fatal adverse effects. PMID- 11067725 TI - Drug-induced cutaneous photosensitivity: some drugs warrant routine precautions. AB - (1) Drug-induced cutaneous photosensitivity is defined as excessive or abnormal skin reactions to light, linked to systemic or topical administration of a drug. (2) The lesions appear after exposure to sunlight or an ultraviolet source, and are limited to or predominate in exposed areas. (3) Some drugs can induce occasionally severe skin burns after moderate exposure to sunlight. Other types of drug-related skin lesion include eczematous or urticarial eruptions, pseudoporphyria, abnormal pigmentation and pseudolichen. Photo-induced or photo aggravated contact dermatitis can occur after application of a photosensitising drug. (4) Some drugs are particularly photosensitising, such as psoralens, tetracyclines, amiodarone and quinolones. (5) Patients treated with drugs known to cause cutaneous photosensitivity should be warned of the risk of cutaneous reactions on exposure to light and be encouraged to protect their skin from intense exposure to sunlight. (6) The possible responsibility of a drug should be borne in mind when a patient presents with skin lesions with a distribution typical of cutaneous photosensitivity. Drug withdrawal and subsequent avoidance can lead to recovery and avoid recurrences. PMID- 11067726 TI - Indications and choice of antihypertensive drugs (cont'd): British recommendations. AB - (1) In the absence of other cardiovascular risk factors, antihypertensive monotherapy should be started in patients with a systolic blood pressure of at least 160 mm Hg or a diastolic pressure of at least 100 mm Hg at repeated measurements. PMID- 11067727 TI - Treatment of tuberculosis: a well-standardised protocol. AB - (1) The recommended treatment protocol for the vast majority of cases of tuberculosis is a 6-month course of isoniazid and rifampicin, plus pyrazinamide and ethambutol for the first two months. (2) This protocol is recommended for both adults and children, and for both pulmonary and extra-pulmonary disease (except for central nervous system involvement). (3) Good adherence to this protocol induces relapse-free recovery in more than 97% of patients with tuberculosis due to non resistant strains. PMID- 11067728 TI - Relations between health professionals and industry: conflicts of interest. AB - Relations between health professionals and drug companies or manufacturers of medical devices can be hazardous. An international debate has developed in recent years in an attempt to limit or, at the very least, elicit "conflicts of interest." PMID- 11067729 TI - Control of communicable diseases; apprehension and detention of persons with specific diseases; transfer of regulations. Food and Drug Administration and Centers for Disease Control and Prevention, HHS. Final rule. AB - The Secretary of Health and Human Service (the Secretary) is transferring a portion of the Food and Drug Administration (FDA) "Control of Communicable Diseases" regulations to the Centers for Disease Control and Prevention (CDC). In general, these regulations provide the Secretary with the authority to apprehend, detain, or conditionally release individuals to prevent the spread of specified communicable diseases. The regulations implement the provisions of the Public Health Service Act (PHS Act) to prevent the introduction, transmission, or spread of communicable diseases from one State or possession into any other State or possession. CDC will have authority for interstate quarantine over persons, while FDA will retain regulatory authority over animals and other products that may transmit or spread communicable diseases. The Secretary is taking this action to consolidate regulations designed to control the spread of communicable diseases, thereby increasing the agencies' efficiency and effectiveness. PMID- 11067730 TI - Health insurance reform: standards for electronic transactions. Office of the Secretary, HHS. Final rule. AB - This rule adopts standards for eight electronic transactions and for code sets to be used in those transactions. It also contains requirements concerning the use of these standards by health plans, health care clearinghouses, and certain health care providers. The use of these standard transactions and code sets will improve the Medicare and Medicaid programs and other Federal health programs and private health programs, and the effectiveness and efficiency of the health care industry in general, by simplifying the administration of the system and enabling the efficient electronic transmission of certain health information. It implements some of the requirements of the Administrative Simplification subtitle of the Health Insurance Portability and Accountability Act of 1996. PMID- 11067731 TI - Revised medical criteria for evaluating mental disorders and traumatic brain injury. Social Security Administration. Final rules. AB - These rules revise our regulations for evaluating mental impairments. They also change some of the provisions of our Listing of Impairments (the Listings) that we use to evaluate mental disorders in adults. We also are adding guidance to the adult neurological listings regarding the evaluation of traumatic brain injury. In addition, the rules make technical changes to the adult digestive listings and the childhood mental disorders listings. We expect that these rules will clarify the intent and purpose of the listings for evaluating mental disorders, and will simplify our adjudication of claims involving mental impairments. These rules also recognize the sometimes unpredictable course of traumatic brain injury, and will improve our adjudication of claims involving traumatic brain injuries. PMID- 11067732 TI - Pediatric testing of prescription drugs: the Food and Drug Administration's carrot and stick for the pharmaceutical industry. PMID- 11067733 TI - Domestic AIDS vaccine trials: addressing the potential for social harm to the subjects of human experiments. AB - In 1998, the FDA approved the first large-scale human trials of a candidate AIDS vaccine in our nation's history. While the legal issues raised by these trials are manifold, the academic literature has focused almost exclusively on the potential for mass tort liability and the resulting hesitancy of biotech and pharmaceutical firms to enter the field. This Comment argues that another issue of vital concern demands attention: the potential for social harm to the human subjects of AIDS vaccine trials. After providing an overview of the current epidemiology of HIV/AIDS and explaining why a safe, effective AIDS vaccine represents the best way to control the pandemic, this Comment analyzes the scientific and social obstacles to production of such a vaccine. In order to know whether a candidate AIDS vaccine is truly effective, researchers will have to test the product in HIV-negative volunteers at high risk of infection. Since these volunteers may subsequently test positive for HIV on standard blood tests, they will be vulnerable to discrimination on that basis in such areas as employment, insurance, immigration, and incarceration. Moreover, by participating in vaccine trials, volunteers will be marking themselves as people at high risk of HIV infection, another basis for disparate treatment. Researchers have suggested that federal disability discrimination law may afford protection against research-related social harms. Through close analysis of the Americans with Disabilities Act of 1990 and the Supreme Court's decision in Bragdon v. Abbott, this Comment demonstrates that optimistic reliance on federal disability law is misplaced. The unique issues raised by domestic AIDS vaccine trials must be addressed in their own right. The Comment accordingly concludes with a broad range of legislative and regulatory proposals to protect trial participants and advance the AIDS vaccine research agenda. PMID- 11067734 TI - Hiding behind agency discretion: the Food and Drug Administration's personal use drug importation policy. PMID- 11067735 TI - Revision of requirements applicable to albumin (human), plasma protein fraction (human), and immune globulin (human). Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is amending the biologics regulations by removing, revising, or updating specific regulations applicable to blood derivative products to be more consistent with current practices and to remove unnecessary or outdated requirements. FDA is taking this action as part of the agency's "Blood Initiative" in which FDA is reviewing and revising, when appropriate, its regulations, policies, guidance, and procedures related to blood products, including blood derivatives. PMID- 11067736 TI - Topical antifungal drug products for over-the-counter human use; amendment of final monograph. Food and Drug Administration, HHS. Final rule. AB - The Food and Drug Administration (FDA) is issuing a final rule amending the monograph for over-the-counter (OTC) topical antifungal drug products. The amendment makes a minor change in the indications for these drug products. This final rule is part of the ongoing review of OTC drug products conducted by FDA. PMID- 11067737 TI - Bonus to reward states for high performance under the TANF program. Administration for Children and Families, HHS. Final rule. AB - The Administration for Children and Families is issuing final regulations to implement section 403(a)(4) of the Social Security Act. This provision authorizes bonuses to high performing States in meeting the purposes of the Temporary Assistance for Needy Families Block Grant (the TANF program). We will base the bonus awards in FY 2002 and beyond on work measures (substantially the same work measures currently in effect for the FY 1999-2001 awards); measures that support work and self-sufficiency related to: participation by low-income working families in the Food Stamp Program, participation of former TANF recipients in the Medicaid and State Children's Health Insurance Programs (SCHIP), and receipt of child care subsidies; and a measure related to family formation and stability (increase in the number of children in the State who reside in married couple families). Bonus funds of up to $200 million each year were authorized for awards in fiscal years 1999 through 2003. This rule specifies a formula for allocating these funds in FY 2002 and FY 2003. The amount awarded to each high performing State may not exceed five percent of the State's family assistance grant. Earlier, we issued program guidance covering bonus awards in FY 1999, FY 2000, and FY 2001. We published a Notice of Proposed Rulemaking to cover awards beginning in FY 2002 on December 6, 1999 (64 FR 68202). In a related regulatory action, we are amending 45 CFR Part 265, the TANF Data Collection and Reporting Requirements, to reduce the burden of reporting data on Separate State Program Maintenance of Effort (SSP-MOE) programs. This amendment will allow waivers of certain reporting requirements under limited circumstances. PMID- 11067738 TI - Studies on the interactions between drugs and estrogen: analytical method for prediction system of gynecomastia induced by drugs on the inhibitory metabolism of estradiol using Escherichia coli coexpressing human CYP3A4 with human NADPH cytochrome P450 reductase. AB - To establish a prediction system for drug-induced gynecomastia in clinical fields, a model reaction system was developed to explain numerically this side effect. The principle is based on the assumption that 50% inhibition concentration (IC(50)) of drugs on the in vitro metabolism of estradiol (E2) to its major product 2-hydroxyestradiol (2-OH-E2) can be regarded as the index for achieving this purpose. By using human cytochrome P450s coexpressed with human NADPH-cytochrome P450 reductase in Escherichia coli as the enzyme, the reaction was examined. Among the nine enzymes (CYP1A1, 1A2, 2A6, 2C8, 2C9, 2C19, 2D6, 2E1, and 3A4) tested, CYP3A4 having a V(max)/K(m) (ml/min/nmol P450) value of 0.32 for production of 2-OH-E2 was shown to be the most suitable enzyme as the reagent. The inhibitory effects of ketoconazole, cyclosporin A, and cimetidine toward the 2-hydroxylation of E2 catalyzed by CYP3A4 were obtained, and their IC(50) values were 7 nM, 64 nM, and 290 microM, respectively. The present results suggest that IC(50) values thus obtained can be substituted as the prediction index for gynecomastia induced by drugs, considering the patients' individual information. PMID- 11067739 TI - The use of protocatechuate dioxygenase for maintaining anaerobic conditions in biochemical experiments. AB - The study of redox-active systems often requires the maintenance of anaerobic conditions. The glucose oxidase system has often been used to maintain anaerobic conditions, but it has some drawbacks, such as the production of H(2)O(2) and limitations on stability. Protocatechuate dioxygenase from Burkholderia cepacia and the substrate, protocatechuate, constitute an alternate effective oxygen scrubbing system that can be used in a wide variety of biochemical experiments. We have shown its suitability for maintaining rigorous anaerobic environments in solutions of pH 6-9, at temperatures from 4 to 35 degrees C, and for periods of time up to 15 months. The enzyme system was shown to be stable under these conditions and effective for maintaining anaerobic conditions in titrations of FAD. It is also suitable for scrubbing various types of apparatus such as stopped flow instruments for anaerobic experiments. PMID- 11067740 TI - The mCK-5 multiprobe RNase protection assay kit can yield erroneous results for the murine chemokines IP-10 and MCP-1. AB - The ribonuclease protection assay (RPA) represents a sensitive method to detect and quantify RNA levels. It can be adapted to allow the simultaneous analysis of more than 10 different mRNAs. The multiprobe RPA kit mCK-5 from PharMingen was used to analyze the expression of chemokines in CCR5 chemokine receptor knockout mice. Upon careful analysis it was found that the mCK-5 kit is defective and can lead to false results for the chemokines IP-10 and MCP-1. The problem is caused by a long-known sequence polymorphism within the 3'-untranslated region of the murine IP-10 gene. This polymorphism leads to a protected IP-10 fragment approximately 20 nucleotides shorter than expected, yielding a length similar to the protected MCP-1 fragment from the mCK-5 kit. Since the identification of specific transcripts with this kit is based exclusively on the size of the various protected fragments, false-negative results for IP-10 together with false positive results for MCP-1 can be obtained. Interestingly, the polymorphism was found not only in 129/CD-1 mice, but also in MRL and SJL/J mice. To facilitate troubleshooting in the future, all templates from the mCK-5 set were isolated and sequenced. PMID- 11067741 TI - Stable expression of varied levels of inducible nitric oxide synthase in primary cultures of endothelial cells. AB - Nitric oxide (NO*), generated by nitric oxide synthase (NOS II) from immunostimulated cells during infection, plays an important role in host immune defense against microbial invasion. The impact of different rates of NO* production on host cell function has not been defined. Herein, we describe the development of a method to express varied levels of murine NOS II in bovine pulmonary artery endothelial cells. A retroviral vector (pMFGSNOS) encoding NOS II was used to transduce primary cultures of endothelial cells. Bovine endothelial cells were susceptible to this transduction and up to 18% of the cells expressed immunodetectable murine NOS II. The NOS II-transduced endothelial cells were cultured on the three-dimensional matrix, Gelfoam, for 8-10 days. Stable expression of NOS II was assessed by measuring nitrite accumulation in media every 2 days. By day 10, endothelial cells on Gelfoam were found to secrete NO* at a rate exceeding 1.0 microM/h/10(6) cells, concomitant with an enhanced level of NOS II activity. Argininosuccinate synthetase, a key enzyme in the metabolism of l-citrulline to l-arginine, increased as well, perhaps in response to dimunition of the intracellular arginine pool corresponding to the observed high output of NO*. In spite of the continuous flux of NO*, endothelial cell viability was not effected. This system provides the opportunity to assess the impact of different levels of sustained NO* production on endothelial cell physiology. PMID- 11067742 TI - Expression of amiloride-sensitive sodium channel: a strategy for the coexpression of multimeric membrane protein in Sf9 insect cells. AB - The amiloride-sensitive epithelial sodium channel (ENaC) mediates Na(+) reabsorption in many epithelial tissues including the distal nephron, colon, lung, and secretory glands and plays an important role in pathophysiology of hypertension and cystic fibrosis. The ENaC is a multimeric integral membrane protein formed by the association of highly homologous,alpha-, beta-, and gamma ENaC subunits. Here we explored the Sf9 insect cell-baculovirus expression system as a source to obtain high yields of recombinant ENaC for functional and structural studies. Although this expression system is widely used, coexpression of ENaC subunits could not be accomplished by the conventional procedures. We thus developed a protocol in which the alpha- and gamma-ENaC cDNA's were first fused individually with polyhedrin promoters at their 5'-ends and then inserted in the multiple cloning sites of pVL1393 transfer vector carrying the beta-ENaC cDNA. Utilizing this transfer vector, a recombinant baculovirus carrying all of the three ENaC cDNA's was prepared. Infection of Sf9 insect cells with this recombinant baculovirus resulted in the expression all of the three ENaC subunits in high yield. Planar lipid bilayer reconstitution procedure revealed the presence of approximately 6 pS sodium channels that are amiloride-sensitive. The results presented point out certain underlying rules for the expression of multiple genes in Sf9 cells, which may be useful in the expression other multimeric proteins and in the studies of protein-protein interactions as well. PMID- 11067743 TI - Measuring the quantity and activity of mitochondrial electron transport chain complexes in tissues of central nervous system using blue native polyacrylamide gel electrophoresis. AB - Mitochondrial dysfunction and degeneration are associated with many neurodegenerative disorders. A dysfunctional mitochondrial electron transport chain (ETC) impairs ATP production and accelerates the generation of free radicals. To evaluate mitochondrial function, reliable methods are needed. Conventional spectrophotometric assays may not eliminate interference from nonspecific enzyme activities and do not measure quantities of specific ETC complexes. Blue native polyacrylamide gel electrophoresis (BN-PAGE) has been used to resolve mitochondrial ETC complexes. Combined with histochemical staining, it has also been applied to measure ETC enzyme activities in muscles. The current study is to determine (1) whether BN-PAGE can be used to detect ETC complexes from different regions of the central nervous system (CNS) and (2) the quantitative range of BN-PAGE in measuring the amounts and activities of different ETC complexes. By systematically varying the protein amount and the time of histochemical reactions, we have found linear ranges comparable to spectrophotometric assays for measuring enzyme activities of several ETC complexes. In addition, we found linear ranges for measuring protein quantities in several ETC complexes. These results demonstrate that BN-PAGE can be used to measure the amount and activity of the ETC enzymes from the nerve tissues and, thus, can be applied to evaluate the functional changes of mitochondria in neurodegenerative disorders. PMID- 11067744 TI - Analysis of purines in urinary calculi by high-performance liquid chromatography. AB - A high-pressure liquid chromatography method has been developed for the analysis in urinary calculi of six purines: uric acid, 2, 8-dihydroxyadenine, xanthine, hypoxanthine, allopurinol, and oxypurinol. Separation was conducted isocratically on a reversed-phase column, using 50 mM phosphate buffer (pH 5.5) / methanol (97/3, v/v) as mobile phase. Limits of detection, depending on compound, ranged from 7 to 28 microg/g stone weight. Hitherto, no reports have appeared on other purines present with uric acid in stones, due to lack of a sensitive and specific analytical method. We have now found that all calculi with more than 4% uric acid also contained 1-methyluric and 7-methyluric acids and trace amounts of hypoxanthine, xanthine, and 2,8-dihydroxyadenine. Accurate identification and quantitation of purines in urinary calculi are important for the diagnosis of rare metabolic diseases leading to urolithiasis (xanthinuria, dihydroxyadeninuria), as well as for prevention of iatrogenic complications during treatment with allopurinol of uric acid urolithiasis. The method may be used for reference purposes in clinical laboratories and for research on the pathogenesis of urolithiasis in disorders of purine metabolism. PMID- 11067745 TI - A high-throughput glow-type aequorin assay for measuring receptor-mediated changes in intracellular calcium levels. AB - A glow-type aequorin luminescence assay for measuring receptor-mediated stimulation of intracellular calcium levels is described and characterized. The human 5-hydroxytryptamine(2A) receptor stably coexpressed in human embryonic kidney cells with apoaequorin was used to characterize the system and showed that following the flash reaction, a stable luminescence signal could be measured using a microplate scintillation counter for between 3 and 7 h after the addition of receptor agonist. Furthermore, this luminescence was dependent on the concentration of agonist used and gave potency values that were stable over this time period. Testing a range of 5-hydroxytryptamine(2A) receptor agonists gave the expected rank order of potency for this receptor. The glow luminescence could also be inhibited by 5-hydroxytryptamine(2A) receptor antagonists, generating affinity values that directly correlated with those determined for inhibition of the flash reaction carried out under the same buffer conditions. The assay therefore gave pharmacologically relevant data and allows a significant improvement of throughput over the traditional flash-type measurements made using an injecting luminometer. PMID- 11067746 TI - Open sandwich enzyme-linked immunosorbent assay for the quantitation of small haptens. AB - The quantitation of low-molecular-weight haptens has been difficult with conventional sandwich immunoassays due to their small size. Many researchers have attempted to develop sandwich assays for haptens due to the significant advantages of the sandwich format over competitive assays including greater dynamic range, ease of automation, and sensitivity. Here we apply the open sandwich ELISA (OS-ELISA), an immunoassay based on antigen-dependent stabilization of antibody variable regions (V(H) and V(L) domains), to hapten quantitation. Two fusion proteins, the high-affinity mutant V(H) domain from anti 4-hydroxy-3-nitrophenacetyl (NP) antibody B1-8 tethered with Escherichia coli alkaline phosphatase (V(H)(W33L)-PhoA) and the V(L) domain from the same antibody tethered with Streptococcus sp. protein G, were made. These fusion proteins when added together achieved Fv reassociation consequent to the addition of NP. Signal was generated in a direct relationship to the NP concentration with better sensitivity compared with competitive immunoassay, demonstrating this assay to be a quick noncompetitive alternative to the conventional assays for small compounds, such as environmental pollutants, drugs of abuse, and therapeutic drugs. With our previous demonstration that the OS-ELISA works well with large proteins, the OS-ELISA becomes the first practical immunoassay approach capable of quantifying any molecule regardless of their size. PMID- 11067747 TI - Photometric method for the quantification of chlorophylls and their derivatives in complex mixtures: fitting with Gauss-peak spectra. AB - Accurate quantification of pigments in mixtures is essential in all cases in which separation of pigments by chromatography is impracticable for one reason or another. An example is the analysis of in vivo formation of heavy metal substituted chlorophylls in heavy metal-stressed plants. We describe here a novel, accurate UV/VIS spectrophotometric method for the quantification of individual chlorophyll derivatives in complex mixtures, which has the potential for universal applicability for mixtures difficult to separate. The method is based on the description of each pigment spectrum by a series of Gaussian peaks. A sample spectrum is then fitted by a linear combination of these "Gauss-peak spectra" including an automatic correction of wavelength inaccuracy and baseline instability of the spectrometer as well as a correction of the widening of absorbance peaks in more concentrated pigment solutions. The automatic correction of peak shifts can also partially correct shifts caused by processes like allomerization. In this paper, we present the Gauss-peak spectra for Mg chlorophyll a, b, c, pheophytin a, b, c, Cu-chlorophyll a, b, c, and Zn chlorophyll a in acetone; Mg-chlorophyll a, b, pheophytin a, b, Cu-chlorophyll a, b, allomerized Cu-chlorophyll a, b, and Zn-chlorophyll a, b in cyclohexane; Mg chlorophyll a, b, pheophytin a, b, and Cu-chlorophyll a, b in diethyl ether. PMID- 11067748 TI - A colorimetric 96-well microtiter plate assay for the determination of enzymatically formed citrulline. AB - l-Citrulline constitutes a product of a number of enzymatic reactions. In the past a number of colorimetric methods for the determination of l-citrulline, upon its chemical modification with diacetyl monoxime at 95 degrees C, have been reported. However, all these methods are time- and material-consuming. In this work, using the same chemical reaction, a new method for the use in 96-well polystyrene microtiter plates was developed. The method is fast and requires substantially less material as the enzymatic reaction is performed in a volume of 60 microl. The applicability of this enzymatic assay was established using l N(omega), N(omega)-dimethylarginine dimethylaminohydrolase, which generates l citrulline from side-chain methylated derivatives of l-arginine. The detection limit for l-citrulline is about 0.2 nmol. In addition, our studies show that most commonly used biochemical buffers and buffer additives do not affect the assay. This method may prove useful in the studies of other l-citrulline producing enzymes including nitric oxide synthase. PMID- 11067749 TI - Full-length single-gene cDNA libraries: applications in splice variant analysis. AB - Alternative splicing of pre-mRNA may generate many distinct proteins from a single gene: regulation of alternative exon selection constitutes control of molecular structure downstream of transcription. Identifying natural splice variants among hundreds or thousands of theoretical alternatives, and examining the regulation of exon selection at multiple sites, may require screening many full-length cDNAs. We describe methods for preparing full-length cDNA libraries comprising the splice variants from single genes. The methods employ robust long distance reverse transcription, gene-specific second strand synthesis, long PCR, and cloning: with these methods cDNAs coding full-length open reading frames were prepared for 21 ion channels (1.2-15 kb). Exon combinations in isolated clones are determined by multiplex PCR. Approximately 85% of the clones contain full length inserts. Screening can detect even rare variants (0.1%) in linear proportion to their abundance in initial mRNA pools. Tissue-specific expression patterns are reproducible. We describe methods for quantifying and minimizing artifactual exon recombination by template switching. These methods can be used to generate thousands of full-length clones of even large transcripts (>8 kb) for the systematic identification of splice variants and the analysis of regulation of alternative exon selection. PMID- 11067750 TI - A HPLC-fluorescence detection method for determination of cardiac phospholipase D activity in vitro. AB - A nonradioactive assay for the investigation of phospholipase D (PLD) activity in cardiac membranes has been developed. A fluorescent derivative of phosphatidylcholine [2-decanoyl-1-(O-(11-(4,4-difluoro-5,7-dimethyl-4-bora-3a, 4a diaza-s-indacene-3proprionyl)amino) undecyl) sn-glycero-3-phosphocholine] was utilized as substrate in an in vitro PLD-catalyzed transphosphatidylation reaction utilizing ethanol as second substrate. Unreacted phosphatidylcholine and the products of phospholipase activity (PEtOH, phosphatidylethanol; PA, phosphatidic acid; DAG, diacylglycerol) were separated by a binary gradient HPLC system and detected by fluorometry. The detection limit of this assay is approximately 0.6 pmol PEtOH. The reaction proceeded at a linear rate for up to 45 min and increased linearly with increasing amounts of rat cardiac membrane protein in a range of 0.625 microg up to at least 25 microg. In the presence of potassium fluoride, formation of fluorescent PA increased at the expense of DAG generation, demonstrating the presence of PA phosphohydrolase activity in rat cardiac membranes. PEtOH formation was unchanged in the presence of the PA phosphohydrolase inhibitor, indicating that the phosphatidylalcohol is not subject to further metabolism by this enzyme. Activation of protein kinase C by phorbol ester significantly increased PLD activity in cardiac membranes. This assay proved to be sensitive for accurate and rapid assessment of PLD activity in cardiac membranes permitting further characterization of the regulation of PLD signal transduction in the heart. PMID- 11067751 TI - Improved performance of pyrosequencing using single-stranded DNA-binding protein. AB - In modern biology, there is a critical need to develop a high-throughput and inexpensive platform for DNA sequencing. Pyrosequencing is a nonelectrophoretic single-tube DNA sequencing method that takes advantage of cooperativity between four enzymes to monitor DNA synthesis. In these studies, single-stranded DNA binding protein (SSB) was added to the primed DNA template prior to the Pyrosequencing reaction. The addition of SSB to a Pyrosequencing reaction system resulted in a read length of more than 30 nucleotides. Improvements were observed as: (i) increased efficiency of the enzymes, (ii) reduced mispriming, as measured by nonspecific signals, (iii) an increase in signal intensity during the reaction, (iv) higher accuracy in reading the number of identical adjacent nucleotides in difficult templates, and (v) longer reads. The usefulness of these results for future Pyrosequencing applications is discussed. PMID- 11067752 TI - A chromogenic substrate for a beta-xylosidase-coupled assay of alpha glucuronidase. AB - 4-Nitrophenyl 2-(4-O-methyl-alpha-d-glucopyranuronosyl)-beta-d-xylopyranoside obtained on deesterification of 4-nitrophenyl 2-O-(methyl 4-O-methyl-alpha-d glucopyranosyluronate)-beta-d-xylopyranoside (Hirsch et al., Carbohydr. Res. 310, 145-149, 1998) was found to be an excellent substrate for the measurement of hemicellulolytic alpha-glucuronidase activity. A new precise alpha-glucuronidase assay was developed by coupling the alpha-glucuronidase-catalyzed formation of 4 nitrophenyl beta-d-xylopyranoside with its efficient hydrolysis by beta xylosidase. A recombinant strain of Saccharomyces cerevisiae, harboring and expressing the beta-xylosidase gene xlnD of Aspergillus niger under control of the alcohol dehydrogenase II promoter on a multicopy plasmid, was used as a source of beta-xylosidase. The activity values of beta-xylosidase in the assay required to achieve a steady-state rate of 4-nitrophenol formation shortly after starting the alpha-glucuronidase reaction were obtained both experimentally and by calculation using the kinetics of coupled enzyme reactions. PMID- 11067753 TI - Peroxidase-active molecular weight markers for direct detection in western blot. PMID- 11067754 TI - A fast method for measurement of branching enzyme activity using a thin-layer chromatography-based phosphorylase a stimulation assay. PMID- 11067755 TI - Factors that influence deoxyribozyme cleavage during polymerase chain reaction. PMID- 11067756 TI - Isolation of rare cDNAs by asymmetric self-hybridization. PMID- 11067757 TI - Enzymatic synthesis of (32)P-labeled phosphoarginine. PMID- 11067758 TI - Thiol alkylation below neutral pH. PMID- 11067759 TI - A single-step, eco-friendly method to extract DNA from Mycobacterium tuberculosis for polymerase chain reaction. PMID- 11067760 TI - A highly active immobilized ribonuclease. PMID- 11067761 TI - Insights on membrane fusion. AB - Membrane fusion is a fundamental cellular process regulating intracellular transport, neurotransmission, enzyme secretion, hormone release, and the entry/exit of viruses, to name a few. Knowledge of how opposing bilayers fuse, besides advancing our understanding of these cellular processes, will provide us with the facts to ameliorate secretory defects and prevent cellular entry or exit of pathogenic viruses. In the last few years, great strides have been made in our understanding of the molecular machinery and mechanism of membrane fusion. In this Special Issue of Cell Biology International, entitled 'Membrane fusion: machinery and mechanism', we have tried to cover several aspects of this vital cellular process, providing insights on the machinery, mechanism and dynamics of the process. Membrane fusion studies reported in this Special Issue have been performed on whole cells, synaptic terminals, viruses, and fusion proteins. PMID- 11067762 TI - KCl stimulation and ultrastructural responses of tannic acid-stained cholinergic synaptic terminals. AB - The quantal-vesicular hypothesis equates miniature end-plate potentials (MEPPs) with fusions of synaptic vesicles. MEPP production thus predicts vesicle losses, increases in vesicle fusions and increases in terminal plasma membrane. MEPP production and these ultrastructural parameters have been evaluated in the cholinergic presynaptic terminals of skate electric organ following tannic acid saline incubation, known to promote capture and selective staining of dense-core granule fusions, and KCl stimulation, known to elevate MEPP production dramatically in these cholinergic terminals. After pretreatment in tannic acid elasmobranch saline, KCl stimulation produced MEPPs at 40/s/microm(2)of terminal surface for several minutes with gradual reduction to spontaneous levels by 25-30 min. No loss of vesicles, no vesicle fusions, no expansions of plasma membrane and no tannic acid enhanced staining of vesicles or vacuoles accompanied the generation of 800 MEPPs/microm(3)of terminals having densities of 567 vesicles/microm(3). No ultrastructural footprints were found to support the notion that unnaturally high rates of vesicular exocytosis had occurred. PMID- 11067763 TI - Fusion of Sendai virus with vesicles of oligomerizable lipids: a microcalorimetric analysis of membrane fusion. AB - Sendai virus fuses efficiently with small and large unilamellar vesicles of the lipid 1,2-di-n-hexadecyloxypropyl-4- (beta-nitrostyryl) phosphate (DHPBNS) at pH 7.4 and 37 degrees C, as shown by lipid mixing assays and electron microscopy. However, fusion is strongly inhibited by oligomerization of the head groups of DHPBNS in the bilayer vesicles. The enthalpy associated with fusion of Sendai virus with DHPBNS vesicles was measured by isothermal titration microcalorimetry, comparing titrations of Sendai virus into (i) solutions of DHPBNS vesicles (which fuse with the virus) and (ii) oligomerized DHPBNS vesicles (which do not fuse with the virus), respectively. The observed heat effect of fusion of Sendai virus with DHPBNS vesicles is strongly dependent on the buffer medium, reflecting a partial charge neutralization of the Sendai F and HN proteins upon insertion into the negatively-charged vesicle membrane. No buffer effect was observed for the titration of Sendai virus into oligomerized DHPBNS vesicles, indicating that inhibition of fusion is a result of inhibition of insertion of the fusion protein into the target membrane. Fusion of Sendai virus with DHPBNS vesicles is endothermic and entropy-driven. The positive enthalpy term is dominated by heat effects resulting from merging of the protein-rich viral envelope with the lipid vesicle bilayers rather than by the fusion of the viral with the vesicle bilayers per se. PMID- 11067764 TI - S100-annexin complexes: some insights from structural studies. AB - Several annexins have been shown to bind proteins that belong to the S100 calcium binding protein family. The two best-characterized complexes are annexin II with p11 and annexin I with S100C, the former of which has been implicated in membrane fusion processes. We have solved the crystal structures of the complexes of p11 with annexin II N-terminus and of S100C with annexin I N-terminus. Using these structural results, as well as electron microscopy observations of liposome junctions formed in the presence of such complexes (Lambert et al., 1997 J Mol Biol 272, 42-55), we propose a computer generated model for the entire annexin II/p11 complex. PMID- 11067765 TI - Persistent fusion pores but transient fusion in alveolar type II cells. AB - The release of vesicle contents following exocytotic fusion is limited by various factors including the size of the fusion pore. Fusion pores are channel-like, narrow structures after formation and proceed through semi-stable states ('fusion pore flickering'), unless they fully expand (full fusion) or close again (transient fusion). Partial release of vesicle contents may occur during transient fusion, which was described to last between milliseconds and seconds, depending on the size of the vesicle. We studied fusion pores in a slow-secreting lung epithelial cell (type II cell) using fluorescence staining of vesicle contents (surfactant) and fluorescence recovery after photobleaching (FRAP). Surfactant is a lipidic material, which is secreted into the alveolar lumen to reduce the surface tension in the lung. We found release of surfactant to be a slow process, which can last for hours. Accordingly, fusion pores in these cells are stable structures, which appear to be a barrier for release. FRAP measurements suggest that transient fusions occasionally take place in these long lasting fusion pores, resulting in partial release of surfactant into the extracellular space. These data suggest that postfusion mechanisms may regulate the amount of secreted surfactant. PMID- 11067766 TI - The t-SNARE syntaxin is sufficient for spontaneous fusion of synaptic vesicles to planar membranes. AB - Vesicular trafficking and exocytosis are directed by the complementary interaction of membrane proteins that together form the SNARE complex. This complex is composed of proteins in the vesicle membrane (v-SNAREs) that intertwine with proteins of the target membrane (t-SNAREs). Here we show that modified synaptic vesicles (mSV), containing v-SNAREs, spontaneously fuse to planar membranes containing the t-SNARE, syntaxin 1A. Fusion was Ca(2+) independent and did not occur with vesicles lacking v-SNAREs. Therefore, syntaxin alone forms a functional fusion complex with v-SNAREs. Our functional fusion assay uses synaptic vesicles that are modified, so each fusion event results in an observable transient current. The mSV do not fuse with protein-free membranes. Additionally, artificial vesicles lacking v-SNAREs do not fuse with membranes containing syntaxin. This technique can be adapted to measure fusion in other SNARE systems and should enable the identification of proteins critical to vesicle-membrane fusion. This will further our understanding of exocytosis and may improve targeting and delivery of therapeutic agents packaged in vesicles. PMID- 11067767 TI - Deployment of membrane fusion protein domains during fusion. AB - It is clear that both viral and intracellular membrane fusion proteins contain a minimal set of domains which must be deployed at the appropriate time during the fusion process. An account of these domains and their functions is given here for the four best-described fusion systems: influenza HA, sendai virus F1, HIV gp120/41 and the neuronal SNARE core composed of synaptobrevin (syn), syntaxin (stx) and the N- and C-termini of SNAP25 (sn25), together with the Ca(2+)binding protein synaptotagmin (syt). Membrane fusion begins with the binding of the virion or vesicle to the target membrane via receptors. The committed step in influenza HA- mediated fusion begins with an aggregate of HAs (at least eight) with some of their HA2 N-termini, a.k.a. fusion peptides, embedded into the viral bilayer (Bentz, 2000 a). The hypothesis presented in Bentz (2000 b) is that the conformational change of HA to the extended coiled coil extracts the fusion peptides from the viral bilayer. When this extraction occurs from the center of the site of restricted lipid flow, it exposes acyl chains and parts of the HA transmembrane domains to the aqueous media, i.e. a hydrophobic defect is formed. This is the 'transition state' of the committed step of fusion. It is stabilized by a 'dam' of HAs, which are inhibited from diffusing away by the rest of the HAs in the aggregate and because that would initially expose more acyl chains to water. Recruitment of lipids from the apposed target membrane can heal this hydrophobic defect, initiating lipid mixing and fusion. The HA transmembrane domains are required to be part of the hydrophobic defect, because the HA aggregate must be closely packed enough to restrict lipid flow. This hypothesis provides a simple and direct coupling between the energy released by the formation of the coiled coil to the energy needed to create and stabilize the high energy intermediates of fusion. Several of these essential domains have been described for the viral fusion proteins SV5 F1 and HIV gp120/41, and for the intracellular SNARE fusion system. By comparing these domains, we have constructed a minimal set which appears to be adequate to explain how the conformational changes can produce a successful fusion event, i.e. communication of aqueous compartments. PMID- 11067768 TI - The secretory pore array hypothesis of transmitter release. PMID- 11067769 TI - Call for the establishment of an international research team for the study of myogenic satellite cells derived from fish. PMID- 11067770 TI - Mechanisms of nongenotoxic carcinogenesis and assessment of the human hazard. AB - Regulatory toxicologists in the pharmaceutical area are faced with many chemical entities to be classified as rodent carcinogens, in most cases on the basis of a nongenotoxic mechanism. The purpose of this paper is to describe some mechanisms for nongenotoxic tumorigenicity and to indicate which type of testing should be done to substantiate why in those cases such a mechanism is not relevant to humans. The increasing attention being given to epigenetic carcinogenesis points at the need for a thorough evaluation during the toxicological program for safety assessment, enabling adequate assessment of the human hazard posed by such compounds. Data to support the nongenotoxic carcinogenesis may be obtained by collecting specific information from current safety assessment programs or from future, separate studies. PMID- 11067771 TI - The utility of PBPK in the safety assessment of chloroform and carbon tetrachloride. AB - Occupational exposure limits (OELs) for individual substances are established on the basis of the available toxicological information at the time of their promulgation, expert interpretation of these data in light of industrial use, and the framework in which they sit. In the United Kingdom, the establishment of specific OELs includes the application of uncertainty factors to a defined starting point, usually the NOAEL from a suitable animal study. The magnitude of the uncertainty factors is generally determined through expert judgment including a knowledge of workplace conditions and management of exposure. PBPK modeling may help in this process by informing on issues relating to extrapolation between and within species. This study was therefore designed to consider how PBPK modeling could contribute to the establishment of OELs. PBPK models were developed for chloroform (mouse and human) and carbon tetrachloride (rat and human). These substances were chosen for examination because of the extent of their toxicological databases and availability of existing PBPK models. The models were exercised to predict the rate (chloroform) or extent (carbon tetrachloride) of metabolism of these substances, in both rodents and humans. Monte Carlo analysis was used to investigate the influence of variability within the human and animal model populations. The ratio of the rates/extent of metabolism predicted for humans compared to animals was compared to the uncertainty factors involved in setting the OES. Predictions obtained from the PBPK models indicated that average rat and mouse metabolism of carbon tetrachloride and chloroform, respectively, are much greater than that of the average human. Application of Monte Carlo analysis indicated that even those people who have the fastest rates or most extensive amounts of metabolism in the population are unlikely to generate the levels of metabolite of these substances necessary to produce overt toxicity in rodents. This study highlights the value that the use of PBPK modeling may add to help inform and improve toxicological aspects of a regulatory process. PMID- 11067772 TI - Safety and advantages of Bacillus thuringiensis-protected plants to control insect pests. AB - Plants modified to express insecticidal proteins from Bacillus thuringiensis (referred to as Bt-protected plants) provide a safe and highly effective method of insect control. Bt-protected corn, cotton, and potato were introduced into the United States in 1995/1996 and grown on a total of approximately 10 million acres in 1997, 20 million acres in 1998, and 29 million acres globally in 1999. The extremely rapid adoption of these Bt-protected crops demonstrates the outstanding grower satisfaction of the performance and value of these products. These crops provide highly effective control of major insect pests such as the European corn borer, southwestern corn borer, tobacco budworm, cotton bollworm, pink bollworm, and Colorado potato beetle and reduce reliance on conventional chemical pesticides. They have provided notably higher yields in cotton and corn. The estimated total net savings to the grower using Bt-protected cotton in the United States was approximately $92 million in 1998. Other benefits of these crops include reduced levels of the fungal toxin fumonisin in corn and the opportunity for supplemental pest control by beneficial insects due to the reduced use of broad-spectrum insecticides. Insect resistance management plans are being implemented to ensure the prolonged effectiveness of these products. Extensive testing of Bt-protected crops has been conducted which establishes the safety of these products to humans, animals, and the environment. Acute, subchronic, and chronic toxicology studies conducted over the past 40 years establish the safety of the microbial Bt products, including their expressed insecticidal (Cry) proteins, which are fully approved for marketing. Mammalian toxicology and digestive fate studies, which have been conducted with the proteins produced in the currently approved Bt-protected plant products, have confirmed that these Cry proteins are nontoxic to humans and pose no significant concern for allergenicity. Food and feed derived from Bt-protected crops which have been fully approved by regulatory agencies have been shown to be substantially equivalent to the food and feed derived from conventional crops. Nontarget organisms exposed to high levels of Cry protein are virtually unaffected, except for certain insects that are closely related to the target pests. Because the Cry protein is contained within the plant (in microgram quantities), the potential for exposure to farm workers and nontarget organisms is extremely low. The Cry proteins produced in Bt-protected crops have been shown to rapidly degrade when crop residue is incorporated into the soil. Thus the environmental impact of these crops is negligible. The human and environmental safety of Bt-protected crops is further supported by the long history of safe use for Bt microbial pesticides around the world. PMID- 11067773 TI - A civil action and statistical assessments of the spatial pattern of disease: do we have a cluster? AB - A reported "cluster" of excess childhood leukemia cases and possible environmental causes in Woburn, Massachusetts, formed a key motivation for the events described in the popular book and motion picture A Civil Action. Although statistical methods to assess spatial clustering existed prior to the events in Woburn, increasing interest in environmental risk factors and recent developments in geographical information systems and data availability prompt increased attention to such methods and their application to public health data. In this article, we review statistical and epidemiological concepts involved in the analysis of disease clusters. We discuss data issues, outline some methodological approaches, and illustrate ideas using data regarding leukemia incidence in upstate New York for the years 1978-1982. PMID- 11067774 TI - The development of glass and stone wool compositions with increased biosolubility. PMID- 11067775 TI - The random-effects model applied to refractory ceramic fiber data. AB - Refractory ceramic fiber (RCF) is a valuable, high-temperature, insulating material with a variety of industrial uses. Because some fibers are respirable by humans and RCF is relatively durable in simulated lung fluids, RCF may pose a health hazard in the workplace. The RCF industry has established a comprehensive product stewardship program (PSP) to identify, quantify, and manage risks. One key element of this PSP is a workplace monitoring program. This paper analyzes monitoring data collected as part of a Consent Agreement with the U.S. Environmental Protection Agency over the period from 1993 to 1998. More specifically, this paper applies the random-effects model (REM) to data collected at several Unifrax plants and applicable to several groups of workers. The REM fits the RCF data well. Depending upon the plant and the functional job category values of the variance of the log-transformed time-weighted average workplace concentrations range from slightly less than 0.5 to 1.0. The estimated intraclass correlations (ratio of the between-worker variance to the total variance) were less than 0.4, and most were less than 0.2. Implications of these findings are examined. Use of the REM in the development of a workplace respiratory policy is described. Finally, two possible criteria for measuring compliance with an occupational exposure limit are reviewed: an "overexposure" criterion developed by Rappaport and co-workers and a conventional "no exceedance" criterion reportedly used by regulatory agencies. The overexposure criterion is logically correct for potential toxicants with chronic effects. For representative values of statistical parameters for RCF from the plants considered, the overexposure criterion is less stringent. PMID- 11067776 TI - Interindividual variance of cytochrome P450 forms in human hepatic microsomes: correlation of individual forms with xenobiotic metabolism and implications in risk assessment. AB - Differences in biotransformation activities may alter the bioavailability or efficacy of drugs, provide protection from certain xenobiotic and environmental agents, or increase toxicity of others. Cytochrome P450 (CYP450) enzymes are responsible for the majority of oxidation reactions of drugs and other xenobiotics and differences in their expression may directly produce interindividual differences in susceptibility to compounds whose toxicity is modulated by these enzymes. To rapidly quantify CYP450 forms in human hepatic microsomes, we developed, and applied, an ELISA to 40 samples of microsomes from adult human organ donors. The procedure was reliable and the results were reproducible within normal limits. Protein content for CYP1A, CYP2E1, and CYP3A positively correlated with suitable marker activities. CYP1A, CYP2B, CYP2C6, CYP2C11, CYP2E1, and CYP3A protein content demonstrated 36-, 13-, 11-, 2-, 12-, and 22-fold differences between the highest and lowest samples and the values were normally distributed. Of the forms examined, CYP3A was expressed in the highest amount and it was the only form whose content was correlated with total CYP450 content. Content of other forms was independent of total CYP450. We further determined the contribution of specific forms to the biotransformation of trichloroethylene as a model substrate. CYP2E1 was strongly correlated with chloral hydrate formation from trichloroethylene; CYP2B displayed the strongest correlation with trichloroethanol formation. These data describing the expression and distribution of these forms in human microsomes can be used to extrapolate in vitro derived metabolic rates for toxicologically important reactions, when form selectivity and specific activity are known. This approach may be applied to refine estimates of human interindividual differences in susceptibility for application in human health risk assessment. PMID- 11067777 TI - Pharmaceutical excipient development: the need for preclinical guidance. AB - Pharmaceutical excipients have a vital role in drug formulations, a role that has tended to be neglected as evidenced by the lack of mechanisms to assess excipient safety outside a new drug application process. Currently, it is assumed that an excipient is "approved" when the new drug formulation, of which it is a constituent, receives regulatory acceptance. Existing regulations and guidelines indicate that new (novel) excipients should be treated as new chemical entities with full toxicological evaluation. No guidance is available for potentially useful materials (essentially new excipients) available from other industries, e.g., food additives or for established excipients with a new application, e.g., dose route change. However, despite this situation, drug companies are actively evaluating new materials or applying new uses to established excipients. Recently developed excipients (e.g., materials giving "sugar-free" status to medical preparations, the cyclodextrins, and the hydrofluoroalkane inhalation propellants) and excipients undergoing development (e.g., chitosan, various enteric coating substances, liposomes, polymers derived from glycolic and lactic acids, and vaccine adjuvants) are all discussed. In light of many other areas of drug development having recently benefited from new or updated regulatory guidance, specific guidance to assist companies in the development of their excipients is urgently needed. Also, an excipient testing strategy would be an excellent topic for inclusion for International Conference on Harmonisation (ICH) consideration. Such guidance/discussion would complement the current advances in pharmacopoeial standardization of excipient quality. As a consequence, it may be possible to have excipients reviewed by a committee of an international pharmacopoeia with the safety data assessed by elected experts and published. PMID- 11067778 TI - The proportions of mutagens among chemicals in commerce. AB - It has been estimated that there are approximately 80,000 chemicals in commerce. Thus, it is not possible to test all these substances for mutagenicity and carcinogenicity; it is possible, however, to test or make estimates from selected subsets of these chemicals. For example, in the U.S. National Toxicology Program (NTP), 35% of the chemicals tested for mutagenicity in Salmonella were positive, as were 52% of the chemicals tested for carcinogenicity in rodents. In contrast, in the U.S. EPA Gene-Tox database, the proportions of chemicals that are Salmonella mutagens is 56%. These and other databases may be biased toward positive responses because they generally have been developed to look at specific structural or use classes of chemicals or chemicals suspected of genetic or carcinogenic activity. To address the question of the proportions of mutagens among all chemicals in commerce, a database of 100 chemicals was created from a random selection of chemicals in commerce. These chemicals were tested for mutagenicity in Salmonella and 22% were mutagenic. The mutagenicity of the 46 highest U.S. production organic chemicals was also compiled; 20% were mutagenic. These values provide a more accurate estimate of the proportions of mutagens among chemicals in commerce than can be derived from published mutagenicity databases. PMID- 11067779 TI - Genomewide search for type 2 diabetes-susceptibility genes in French whites: evidence for a novel susceptibility locus for early-onset diabetes on chromosome 3q27-qter and independent replication of a type 2-diabetes locus on chromosome 1q21-q24. AB - Despite recent advances in the molecular genetics of type 2 diabetes, the majority of susceptibility genes in humans remain to be identified. We therefore conducted a 10-cM genomewide search (401 microsatellite markers) for type 2 diabetes-related traits in 637 members of 143 French pedigrees ascertained through multiple diabetic siblings, to map such genes in the white population. Nonparametric two-point and multipoint linkage analyzes-using the MAPMAKER-SIBS (MLS) and MAXIMUM-BINOMIAL-LIKELIHOOD (MLB) programs for autosomal markers and the ASPEX program for chromosome X markers-were performed with six diabetic phenotypes: diabetes and diabetes or glucose intolerance (GI), as well as with each of the two phenotypes associated with normal body weight (body-mass index<27 kg/m(2)) or early age at diagnosis (<45 years). In a second step, high-resolution genetic mapping ( approximately 2 cM) was performed in regions on chromosomes 1 and 3 loci showing the strongest linkage to diabetic traits. We found evidence for linkage with diabetes or GI diagnosed at age <45 years in 92 affected sib pairs from 55 families at the D3S1580 locus on chromosome 3q27-qter using MAPMAKER-SIBS (MLS = 4.67, P=.000004), supported by the MLB statistic (MLB LOD=3.43, P=.00003). We also found suggestive linkage between the lean diabetic status and markers APOA2-D1S484 (MLS = 3. 04, P=.00018; MLB-LOD=2.99, P=.00010) on chromosome 1q21-q24. Several other chromosomal regions showed indication of linkage with diabetic traits, including markers on chromosome 2p21-p16, 10q26, 20p, and 20q. These results (a) showed evidence for a novel susceptibility locus for type 2 diabetes in French whites on chromosome 3q27-qter and (b) confirmed the previously reported diabetes-susceptibility locus on chromosome 1q21-q24. Saturation on both chromosomes narrowed the regions of interest down to an interval of <7 cM. PMID- 11067780 TI - Primary autosomal recessive microcephaly: MCPH5 maps to 1q25-q32. AB - Primary microcephaly is thought to result from genetic defects of the developmental program that generates large brain hemispheres in humans. Autosomal recessive inheritance is likely in most familial cases, and four loci were recently mapped by homozygosity. We report homozygosity mapping of a new locus, MCPH5, with a maximum multipoint LOD score of 3.51 at marker D1S1723, in a family of Turkish origin. The minimal critical region spans 11.4 cM between markers D1S384 and D1S2655, at 1q25-q32, and encompasses the cytogenetic breakpoints of chromosomal aberrations previously reported in unrelated patients with microcephaly. PMID- 11067781 TI - Genetics of prostate cancer: too many loci, too few genes. PMID- 11067782 TI - Photoangioplasty for human peripheral atherosclerosis: results of a phase I trial of photodynamic therapy with motexafin lutetium (Antrin). AB - BACKGROUND: In photoangioplasty, light activation of a photosensitive drug offers the potential for treatment of long segments of vascular disease. This is a brief description of a study designed to evaluate the safety and tolerability of a new photosensitizer, Antrin (motexafin lutetium), in the endovascular treatment of atherosclerosis. METHODS AND RESULTS: An open-label, single-dose, escalating drug and light-dose study was performed in patients with atherosclerotic peripheral arterial insufficiency. Clinical evaluation, serial quantitative angiography, and intravascular ultrasonography were performed. Therapy was well tolerated, and only minor side effects were observed. Treatment produced no deleterious vascular effects. Although this study was not designed to examine clinical efficacy, several secondary end points suggested a favorable therapeutic effect. CONCLUSIONS: This phase I study demonstrates that photoangioplasty with motexafin lutetium is well tolerated and safe. Preliminary efficacy data suggest a future role for the treatment of flow-limiting atherosclerosis. PMID- 11067783 TI - Striking increase of natriuresis by low-dose spironolactone in congestive heart failure only in combination with ACE inhibition: mechanistic evidence to support RALES. AB - BACKGROUND: A marked reduction of overall mortality in patients with severe congestive heart failure (CHF) has been demonstrated by addition of the mineralocorticoid receptor antagonist spironolactone to ACE inhibition. The aim of the present study was to examine a hypothesized interaction of spironolactone and ACE inhibitors in renal electrolyte and volume regulation. METHODS AND RESULTS: Wistar rats with extensive myocardial infarction or sham operation were treated with either placebo, the ACE inhibitor trandolapril, low-dose spironolactone, or a combination of the 2. Twelve weeks after infarction, rats were housed in metabolic cages. Urinary volume and sodium excretion were significantly increased in CHF rats on a combined treatment with spironolactone and trandolapril (21.2+/-2.6 mL/d, 2489+/-320 mmol/d, mean+/-SD; P<0.05 versus other experimental groups) versus placebo-treated rats (16.7+/-5.6 mL/d, 1431+/ 458 mmol/d),whereas these parameters were not affected in rats on either spironolactone (16.1+/-6.6 mL/d, 1153+/-273 mmol/d) or trandolapril alone (15.9+/ 4.2 mL/d, 1392+/-294 mmol/d). The effects on natriuresis coincided with a significant reduction of left ventricular end-diastolic pressure (LVEDP) in rats on trandolapril and spironolactone (10.8+/-8.2 mm Hg; P:<0.05 versus CHF placebo: 23.3+/-7.2 mm Hg; sham-operated rats: 5.1+/-0.9 mm Hg), whereas LVEDP remained elevated in rats treated with either compound alone. CONCLUSIONS: In the present study, we found an unexpected interaction of low-dose spironolactone and the ACE inhibitor trandolapril in experimental CHF leading to marked effects on renal electrolyte and volume regulation that were not apparent by treatment with either drug alone. These findings may explain the beneficial effects of spironolactone in CHF patients. PMID- 11067784 TI - Association between white blood cell count, epicardial blood flow, myocardial perfusion, and clinical outcomes in the setting of acute myocardial infarction: a thrombolysis in myocardial infarction 10 substudy. AB - BACKGROUND: Elevation of the white blood cell (WBC) count during acute myocardial infarction (AMI) is associated with adverse outcomes. We examined the relationship between the WBC count and angiographic findings to gain insight into this relationship. Results and Methods-We evaluated data from 975 patients in the Thrombolysis In Myocardial Infarction (TIMI) 10A and 10B trials. Patients with a closed artery at 60 and 90 minutes had higher a WBC count than patients with an open artery (P:=0.02). Likewise, the presence of angiographically apparent thrombus was associated with a higher WBC count (11.5+/-5.2x10(9)/L, n=290, versus 10.7+/-3. 5x10(9)/L, n=648; P=0.008). In addition, a higher WBC count was associated with poorer TIMI myocardial perfusion grades (4-way P=0.04). Mortality rates were higher in patients with a higher WBC count (0% for WBC count 0 to 5x10(9)/L, 4.9% for WBC count 5 to 10x10(9)/L, 3.8% for WBC count 10 to 15x10(9)/L, 10.4% for WBC count >15x10(9)/L; P=0.03). The development of new congestive heart failure or shock was also associated with a higher WBC count (0% for WBC count 0 to 5x10(9)/L, 5.2% for WBC count 5 to 10x10(9)/L, 6.1% for WBC count 10 to 15x10(9)/L, 17.1% for WBC count >15x10(9)/L; P<0.001), an observation that remained significant in a multivariable model that adjusted for potential confounding variables (odds ratio 1.21, P=0.002). CONCLUSIONS: Elevation in WBC count was associated with reduced epicardial blood flow and myocardial perfusion, thromboresistance (arteries open later and have a greater thrombus burden), and a higher incidence of new congestive heart failure and death. These observations provide a potential explanation for the higher mortality rate observed among AMI patients with elevated WBC counts and helps explain the growing body of literature that links inflammation and cardiovascular disease. PMID- 11067785 TI - Chlamydia pneumoniae, herpes simplex virus type 1, and cytomegalovirus and incident myocardial infarction and coronary heart disease death in older adults : the Cardiovascular Health Study. AB - BACKGROUND: Whether serological evidence of prior infection with Chlamydia pneumoniae, herpes simplex virus type 1 (HSV-1), and cytomegalovirus (CMV) is associated with myocardial infarction (MI) and coronary heart disease (CHD) death remains a source of controversy. METHODS AND RESULTS: We conducted a nested case control study among participants in the Cardiovascular Health Study, a cohort study of persons aged >/=65 years. Cases experienced an incident MI and CHD death (n=213). Control subjects were matched to cases by age, sex, clinic, year of enrollment, and month of blood draw (n=405). Serum was analyzed for IgG antibodies to C pneumoniae, HSV-1, and CMV. After adjustment for other risk factors, the risk of MI and CHD death was associated with the presence of IgG antibodies to HSV-1 (odds ratio [OR] 2.0, 95% CI 1.1 to 3.6) but was not associated with the presence of IgG antibodies to either C pneumoniae (OR 1.1, 95% CI 0.7 to 1.8) or CMV (OR 1.2, 95% CI 0.7 to 1.9). Although there was little association with low to moderate C pneumoniae antibody titers (0.26 micromol/L with at least 50% reduction after cobalamin supplementation. ROC and logistic regression analyses were used. RESULTS: Serum cobalamin and tHcy were the best predictors, with areas under the ROC curve (SE) of 0. 810 (0.034) and 0.768 (0.037), respectively, but age, intrinsic factor antibodies, and gastroscopy gave additional information. CONCLUSIONS: When cobalamin deficiency is suspected in general practice, serum cobalamin should be the first diagnostic test, and the result should be interpreted in relation to the age of the patient. When a definite diagnosis cannot be reached, MMA and tHcy determination will provide additional discriminative information, but MMA, being more specific, is preferable for assessment of cobalamin status. PMID- 11067809 TI - Rapid screening method for osteoclast differentiation in vitro that measures tartrate-resistant acid phosphatase 5b activity secreted into the culture medium. AB - BACKGROUND: Osteoclasts secrete tartrate-resistant acid phosphatase (TRAP; EC 3.1.3.2) 5b into the circulation. We studied the release of TRAP 5b from osteoclasts using a mouse in vitro osteoclast differentiation assay. METHODS: We developed and characterized a polyclonal antiserum in rabbits, using purified human osteoclastic TRAP 5b as antigen. The antiserum was specific for TRAP in Western analysis of mouse osteoclast culture medium and was used to develop an immunoassay. We cultured mouse bone marrow-derived osteoclast precursor cells for 3-7 days with or without clodronate in the presence of vitamin D and analyzed the number of osteoclasts formed and the amount of TRAP 5b activity released into the culture medium. RESULTS: TRAP 5b activity was not secreted from osteoclast precursor cells. Addition of clodronate-containing liposomes decreased in a dose dependent manner the number of osteoclasts and TRAP 5b activity released in 6-day cultures. The amount of TRAP 5b activity in the medium detected by the immunoassay correlated significantly with the number of osteoclasts formed (r = 0.94; P<0.0001; n = 120). CONCLUSIONS: The TRAP 5b immunoassay can be used to replace the laborious and time-consuming microscopic counting of osteoclasts in the osteoclast differentiation assay and to test the effects of potential therapeutic agents on osteoclast differentiation, enabling fast screening of large amounts of potential therapeutic agents. PMID- 11067810 TI - Miniature single-particle immunoassay for prostate-specific antigen in serum using recombinant Fab fragments. AB - BACKGROUND: Quantitative, miniaturized nucleic acid assays and immunoassays can be developed with single microparticles, microfluorometric detection, and intrinsically fluorescent lanthanide chelates in a multiple assay format to decrease reagent consumption, cost, and assay time. We used recombinant Fab fragments to capture and detect free and total prostate-specific antigen (PSA) from serum in a submicroliter volume single-particle immunoassay. METHODS: Genetically engineered thiol-Fab or thiolated monoclonal antibodies (mAbs) were covalently attached onto uniformly sized 60-microm maleimide-activated microparticles. Free and total PSA were detected with europium- or terbium labeled Fab fragments on a single microparticle using a microfluorometer in a time-resolved mode. RESULTS: The detection limit of the free- and total-PSA assays (mean + 3 SD of zero calibrator) was 0.35 microg/L, with a total volume of 330 nL per particle. An excellent correlation was found in microparticle and microtiter-well assays for 21 serum samples: slopes for free and total PSA were 1.06+/-0.03 and 1.03+/-0.02, respectively (S(y|x) = 0.084 and 0.057 microg/L), with intercepts of 0.013+/-0.018 and 0.013+/-0.017 microg/L (R>0.99). Furthermore, the particle-immobilized Fab fragment had a PSA binding capacity 1.5 fold higher than the intact mAb capacity on a single microparticle. Capacity, kinetics, and sensitivity of the Fab fragment and intact mAb assays in the microparticle and microtiter well formats are discussed. CONCLUSIONS: With site specific (cysteine tail) covalent attachment of Fab fragments on a microparticle, subattomole amounts of PSA can be detected quantitatively. PMID- 11067811 TI - A reference method laboratory network for cholesterol: a model for standardization and improvement of clinical laboratory measurements. AB - BACKGROUND: Accurate and precise measurement of blood cholesterol plays a central role in the National Cholesterol Education Program's strategy to reduce the morbidity and mortality attributable to coronary heart disease. Matrix effects hamper the ability of manufacturers to adequately calibrate and validate traceability to the National Reference System for Cholesterol (NRS/CHOL). CDC created the Cholesterol Reference Method Laboratory Network (CRMLN) to improve cholesterol measurement by assisting manufacturers of in vitro diagnostic products with validation of the traceability of their assays to the NRS/CHOL. METHODS: CRMLN laboratories established the CDC cholesterol reference method (modification of the Abell-Levy-Brodie-Kendall chemical method) and are standardized using CDC frozen serum reference materials. CRMLN laboratories use common quality-control materials and participate in monthly external performance evaluations conducted by CDC. The CRMLN performance criteria require member laboratories to agree with CDC within +/-1.0% and maintain a CV < or =2.0%. RESULTS: From 1995 to 200 the CRMLN laboratories met the accuracy criterion 97% of the time and the precision criterion 99% of the time. During this time period, the CRMLN maintained an average bias to CDC of 0.01% and an average collective CV of 0.33%. CONCLUSIONS: CDC established the CRMLN as the first international reference method laboratory network. The CRMLN assists manufacturers in the validation of the calibration of their diagnostic products so that clinical laboratories can measure blood cholesterol more reliably. The CRMLN can serve as a model for other clinical analytes where traceability to a hierarchy of methods is needed and matrix effects of the field methods with processed calibrators or reference materials are present. PMID- 11067812 TI - Comparison of capillary electrophoresis with HPLC for diagnosis of factitious hypoglycemia. AB - BACKGROUND: The diagnosis of "factitious hypoglycemia" is essentially based on the disclosure of hypoglycemic agents in blood or urine. The aim of this study was to evaluate the performance of capillary electrophoresis (CE) as a quantitative method for determination of chlorpropamide, tolbutamide, glipizide, gliclazide, and glibenclamide in serum. METHODS: Serum samples (1 mL), with internal standard added, were purified by solid-phase extraction on OASIS(TM) HLB cartridges (Waters), dried under reduced pressure, and reconstituted with 30-60 microL of acetonitrile:H(2)O. Analysis was carried out by micellar electrokinetic capillary chromatography in 5 mmol/L borate, 5 mmol/L phosphate, 75 mmol/L sodium cholate, pH 8.5, containing 25 mL/L methanol. Separation was accomplished in a 20 cm x 50 microm (i.d.) silica capillary at 25 degrees C and a constant voltage of +10 kV. Pharmacokinetics of gliclazide (80-mg tablet) in a diabetic patient were assayed by both HPLC and CE. Two hypoglycemic patients positive by HPLC analysis for unreported gliclazide and tolbutamide overdose were also screened by CE. RESULTS: Separation of six drugs (including the internal standard) was accomplished in 5 min plus 5 min rinsing. The between-day CV of the ratio of the areas of the sulfonylurea drugs to internal standard was <1% (n = 10). Linearity (r(2) > or =0.998) and recovery (> or =80%) were good for all sulfonylurea drugs tested. Pharmacokinetic curves for gliclazide by CE and HPLC were superimposable. CE analysis confirmed the HPLC diagnosis of surreptitious abuse of gliclazide and tolbutamide. CONCLUSION: CE is a useful tool in the clinical chemistry and toxicology laboratory for drug monitoring and pharmacokinetic investigations. PMID- 11067813 TI - Human urine certified reference material for arsenic speciation. AB - BACKGROUND: Chemical speciation analysis is essential for the biological monitoring of inorganic arsenic exposure using urine as indicator medium. There is increasing demand for a certified reference material (CRM) of urine matrix for arsenic speciation. METHODS: Urine (10 L) was collected from non-occupationally exposed Japanese males. We prepared 954 bottles of urine, each containing approximately 10 mL, after filtering and blending the urine stock. The urine in each bottle was freeze-dried. Between-bottle homogeneity was confirmed by measuring the concentrations of selected minor and trace elements in the material and subsequent statistical analysis. Certification was based on a collaborative analysis involving 15 laboratories. RESULTS: Certified values were determined for arsenobetaine (0.069+/-0.012 mg As/L), dimethylarsinic acid (0.036+/-0.009 mg As/L), and total arsenic (0.134+/-0.011 mg/L) as well as for total selenium (0.059+/-0.005 mg/L) and zinc (0.62+/-0.05 mg/L), based on the analytical values from the collaborating laboratories. Reference values are given for copper (0.010 mg/L) and lead (0.0011 mg/L), based on definitive analysis at the National Institute for Environmental Studies (NIES). CONCLUSIONS: The present CRM, NIES CRM No. 18 Human Urine, is the first human urine CRM for arsenic speciation and will be of value for analytical quality assurance of the biological monitoring of arsenic exposure. PMID- 11067814 TI - Stability of methylecgonidine and ecgonidine in sheep plasma in vitro. AB - BACKGROUND: Crack smokers are exposed to a pyrolysis product, methylecgonidine (MEG), which can be used as an analytical marker for crack smoking. Ecgonidine (EC), a hydrolytic product of MEG, has been identified in urine of crack smokers. MEG undergoes conversion to EC, complicating analysis and perhaps explaining a lack of forensic blood specimens containing MEG. METHODS: We developed gas chromatography-mass spectrometry (GC-MS) assays for MEG and EC. Plasma was collected from sheep blood containing 0, 0.06, or 0.24 mol/L (0%, 0.25%, or 1%) NaF. MEG was added to these plasmas, and they were incubated at -80, 1, 21, or 37 degrees C to determine whether there were temporal, temperature, or storage effects on MEG stability over 48 h. RESULTS: Decreased temperature and increased NaF concentrations limited MEG degradation and EC formation. MEG stored in plasma at -80 degrees C was stable up to 1 month, even in the absence of NaF. CONCLUSIONS: MEG is stable in sheep plasma collected in commercially available, evacuated blood-collection tubes containing NaF and stored at -80 degrees C. In vitro formation of EC can be minimized with appropriate sample handling, and its in vivo formation may provide a better marker of crack smoking than its parent pyrolysis product. PMID- 11067815 TI - Determination of the glycoforms of human chorionic gonadotropin beta-core fragment by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - BACKGROUND: Metabolism of human chorionic gonadotropin (hCG) in the serum and kidney yields the terminal urinary product hCG beta-core fragment (hCGbetacf), comprising two disulfide-linked peptides (beta6-beta40 and beta55-beta92) of which one (beta6-beta40) retains truncated N-linked sugars. Hyperglycosylated hCGbetacf may indicate choriocarcinoma or Down syndrome, but the glycosylation profile of hCGbetacf has not been thoroughly evaluated. METHODS: hCGbetacf, purified from pregnancy urine, was reduced by "on-target" dithiothreitol (DTT) reduction and analyzed by matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS). The mass ([M+H](+)) of the primary sequence of the glycosylated peptide beta6-beta40 was subtracted from the m/z values of the discrete peaks observed to give the masses of the carbohydrate moieties. Carbohydrate structure was predicted by sequentially subtracting the masses of the monosaccharide residues corresponding to N-linked carbohydrates of the hCG beta-subunit reported in the literature. RESULTS: Mass spectra of hCGbetacf revealed a broad triple peak at m/z 8700-11300. After reduction, the triple peak was replaced by a discrete set of peaks between m/z 4156 and 6354. A peak at m/z 4156.8 corresponded to the nonglycosylated peptide (beta55-beta92). The remaining nine peaks indicated that urinary hCGbetacf comprises a set of glycoforms smaller and larger than the trimannosyl core. CONCLUSIONS: hCGbetacf comprises a wider set of glycoforms than reported previously. Peaks of highest mass indicate evidence of hyperglycosylated carbohydrate moieties. The data support previous reports that hCGbetacf oligosaccharides lack sialic acid and galactose residues. No indication was found of a beta6-beta40 peptide that was entirely devoid of carbohydrate. PMID- 11067816 TI - Methylmalonic acid measured in plasma and urine by stable-isotope dilution and electrospray tandem mass spectrometry. AB - BACKGROUND: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionization is robust and allows accurate measurement of both low- and high-molecular weight components of complex mixtures. We developed a LC-MS/MS method for the analysis of methylmalonic acid (MMA), a biochemical marker for inherited disorders of propionate metabolism and acquired vitamin B(12) deficiency. METHODS: We added 1 nmol of the internal standard MMA-d(3) to 500 microL of plasma or 100 microL of urine before solid-phase extraction. After elution with 18 mol/L formic acid, the eluate was evaporated, and butyl ester derivatives were prepared with 3 mol/L HCl in n-butanol at 65 degrees C for 15 min. For separation, we used a Supelcosil LC-18, 33 x 4.6 mm column with 60:40 (by volume) acetonitrile:aqueous formic acid (1 g/L) as mobile phase. The transitions m/z 231 to m/z 119 and m/z 234 to m/z 122 were used in the selected reaction monitoring mode for MMA and MMA-d(3,) respectively. The retention time of MMA was 2.2 min in a 3.0-min analysis, without interference of a physiologically more abundant isomer, succinic acid. RESULTS: Daily calibrations between 0.25 and 8.33 nmol in 0.5 mL exhibited consistent linearity and reproducibility. At a plasma concentration of 0.12 micromol/L, the signal-to noise ratio for MMA was 40:1. The regression equation for our previous gas chromatography-mass spectrometry (GC-MS) method (y) and the LC-MS/MS method (x) was: y = 1.030 x -0.032 (S(y|x) = 1.03 micromol/L; n = 106; r = 0.994). Inter- and intraassay CVs were 3. 8-8.5% and 1.3-3.4%, respectively, at mean concentrations of 0.13, 0.25, 0.60, and 2.02 micromol/L. Mean recoveries of MMA added to plasma were 96.9% (0.25 micromol/L), 96.0% (0.60 micromol/L), and 94.8% (2.02 micromol/L). One MS/MS system used only overnight (7.5 h) replaced two GC MS systems (30 instrument-hours/day) to run 100-150 samples per day, with reductions of total cost (supplies plus equipment), personnel, and instrument time of 59%, 14%, and 75%, respectively. CONCLUSIONS: This method is well suited for large-scale MMA testing (> or =100 samples per day) where a shorter analytical time is highly desirable. Reagents are less expensive than the anion exchange/cyclohexanol-HCl method, and sample preparation of batches up to 100 specimens is completed in less than 8 h and is automated. PMID- 11067817 TI - Evaluation of LabRespond, a new automated validation system for clinical laboratory test results. AB - BACKGROUND: Manual validation of laboratory test results is time-consuming, creating a demand for expert systems to automate this process. We have started to set up the program "LabRespond", which covers five validation levels: administrative, technical, sample, patient, and clinical validation. We present the evaluation of a prototype of an automated patient validation system based on statistical methods, in contrast to the commercially available program "VALAB", a rule-based automated validation system. METHODS: In the present study, 163 willfully altered, erroneous test results out of 5421 were submitted for validation to LabRespond, VALAB, and to a group of clinical chemists (n = 9) who validated these test results manually. The test results rejected by three or more clinical chemists (n = 281) served as a secondary reference standard. RESULTS: The error recovery rates of clinical chemists ranged from 23.9% to 71.2%. The recovery rates of LabRespond and VALAB were 77.9% and 71.8%, respectively (difference not significant). The false-positive rates were 82.7% for LabRespond, 83.6% for VALAB, and 27.8-86.7% for clinical chemists. Using the consensus of three or more clinical chemists as the secondary reference standard, we found error recovery rates of 64.8% for LabRespond and 72.2% for VALAB (P = 0.06). Compared with VALAB, LabRespond detected more (P = 0.003) erroneous test results of the type that were changed from abnormal to normal. CONCLUSIONS: The statistical plausibility check used by LabRespond offers a promising automated validation method with a higher error recovery rate than the clinical chemists participating in this study, and a performance comparable to VALAB. PMID- 11067818 TI - Influence of increased fruit and vegetable intake on plasma and lipoprotein carotenoids and LDL oxidation in smokers and nonsmokers. AB - BACKGROUND: Epidemiological studies suggest a cardioprotective role for carotenoid-rich foods. Smokers have a high risk of cardiovascular disease and low dietary intake and plasma concentrations of carotenoids. The aim of this study was to determine the carotenoid response of smokers and nonsmokers to increased intake of 300-400 g of vegetables and its effect on LDL oxidation. METHODS: After a depletion period of 8 days, 34 healthy females (18 nonsmokers, 16 smokers) were supplemented with beta-carotene- and lutein-rich (green) and lycopene-rich (red) vegetable foods, each for 7 days. RESULTS: Baseline concentrations (mean +/- SD) of plasma beta-carotene (0.203+/-0.28 micromol/L vs. 0.412+/-0.34 micromol/L; P <0.005) and lutein (0.180 +/-0.10 vs. 0.242+/-0.11 micromol/L; P<0.05) but not lycopene (0.296+/-0.10 vs. 0.319+/-0.33 micromol/L) were significantly lower in smokers compared with nonsmokers. After supplementation, the change (supplementation minus depletion) in plasma beta-carotene (0.152+/- 0.43 vs. 0.363+/-0.29 micromol/L in smokers vs. nonsmokers; P = 0.002) and LDL lutein (0.015+/-0.03 vs. 0.029+/-0.03 micromol/mmol cholesterol; P = 0.01) was significantly lower in smokers than nonsmokers. Green-vegetable supplementation had no effect on the resistance of LDL to oxidation (lag-phase) in either group. After red-vegetable supplementation, plasma and LDL lycopene concentrations were increased in both groups, but only nonsmokers showed a significant increase in the lag-phase (44.9+/-9.5 min at baseline, 41.4+/-6.5 min after depletion, and 49.0+/-8.9 min after supplementation; P<0.01) compared with depletion. CONCLUSIONS: In this short-term intervention study, a dietary intake of >40 mg/day of lycopene by a group of nonsmoking individuals significantly reduced the susceptibility of LDL to oxidation, whereas an equivalent increase in lycopene by a group of smokers showed no such effect. PMID- 11067819 TI - Inaccuracy of calculated LDL-cholesterol in type 2 diabetes: consequences for patient risk classification and therapeutic decisions. PMID- 11067820 TI - Presence of fetal RNA in maternal plasma. PMID- 11067821 TI - CYP3A4-V polymorphism detection by PCR-restriction fragment length polymorphism analysis and its allelic frequency among 199 Dutch Caucasians. PMID- 11067822 TI - Phagocytosis and oxidative burst: reference values for flow cytometric assays independent of age. PMID- 11067823 TI - Development and validation of an automated and ultrasensitive immunoturbidimetric assay for C-reactive protein. PMID- 11067824 TI - Flexibility of melting temperature assay for rapid detection of insertions, deletions, and single-point mutations of the AGXT gene responsible for type 1 primary hyperoxaluria. PMID- 11067825 TI - Measurement of prothrombin time in EDTA plasma with combined thromboplastin reagent. PMID- 11067826 TI - Stability of cannabinoids in hair samples exposed to sunlight. PMID- 11067827 TI - Evaluation of a homogeneous direct LDL-cholesterol assay in diabetic patients: effect of glycemic control. PMID- 11067828 TI - Evaluation of DNA fragment sizing and quantification by the agilent 2100 bioanalyzer. PMID- 11067829 TI - Evaluation of a nucleic acid-based cross-linking assay to screen for hereditary hemochromatosis in healthy blood donors. PMID- 11067830 TI - New nomenclature for the human tissue kallikrein gene family. PMID- 11067831 TI - Association of a missense Glu298Asp mutation of the endothelial nitric oxide synthase gene with end stage renal disease. PMID- 11067832 TI - Presence of birefringent, maltese-cross-appearing spherules in synovial fluid in a case of acute monoarthritis. PMID- 11067833 TI - Temperature variations in chest-type mechanical freezers. PMID- 11067834 TI - Soluble Fas in serum of patients with HIV/AIDS. PMID- 11067835 TI - Comparison of cardiac troponin I measurements on whole blood and plasma on the Stratus CS analyzer and comparison with AxSYM. PMID- 11067836 TI - Real-time PCR assay with fluorescent hybridization probes for rapid genotyping of the CD14 promotor polymorphism. PMID- 11067837 TI - Assessment of vitamin B(1) status. PMID- 11067838 TI - Transient hyperphosphatasemia of infancy and childhood: study of 194 cases. PMID- 11067839 TI - Comparison of single and repeat centrifugation of blood specimens collected in BD evacuated blood collection tubes containing a clot activator for cardiac troponin I assay on the ACCESS analyzer. PMID- 11067840 TI - Unsaturated iron-binding capacity: a screening test for C282Y hemochromatosis? PMID- 11067841 TI - The San Diego Conference. The current revolution: SNPs and chips in molecular diagnostics. November 16-18, 2000. Abstracts. PMID- 11067842 TI - Compiled by david E. Bruns, editor (dbruns@aacc.org) PMID- 11067843 TI - Characterization of a novel trans-activation domain of BRCA1 that functions in concert with the BRCA1 C-terminal (BRCT) domain. AB - Mutations in the breast cancer susceptibility gene, BRCA1, account for a significant proportion of hereditary breast and ovarian cancers. The BRCA1 C terminal (BRCT) domain, which can activate transcription when fused to a heterologous DNA binding domain, is required for BRCA1 function in suppression of tumorigenesis. Here, we provide evidence for a new activation domain in BRCA1 that lies adjacent to the BRCT domain. We name the two domains AD1 and AD2, respectively. Like AD2, the newly discovered AD1 can act independently as an activation domain in both yeast and human cells. However, unlike AD2, AD1 activity in mammalian cells is cell type context-dependent. Furthermore, combination of these two domains in mammalian cells can result in a robust synergy in transcriptional activation. A highly conserved coiled-coil motif in AD1 is required for the cooperative transcription activation. Interestingly, the functional cooperativity between AD1 and AD2 is absent in certain breast and ovarian cancer cell lines, although each domain can still activate transcription. Therefore, the differential and cooperative actions of the two activation modules may contribute to the heterogeneous risk of BRCA1 mutations in different tissues. PMID- 11067844 TI - A bipartite substrate recognition motif for cyclin-dependent kinases. AB - Cy or RXL motifs have been previously shown to be cyclin binding motifs found in a wide range of cyclin-Cdk interacting proteins. We report the first kinetic analysis of the contribution of a Cy motif on a substrate to phosphorylation by cyclin-dependent kinases. For both cyclin A-Cdk2 and cyclin E-Cdk2 enzymes, the presence of a Cy motif decreased the K(m(peptide)) 75-120-fold while the k(cat) remained unchanged. The large effect of the Cy motif on the K(m(peptide)) suggests that the Cy motif and (S/T)PX(K/R) together constitute a bipartite substrate recognition sequence for cyclin-dependent kinases. Systematic changes in the length of the linker between the Cy motif and the phosphoacceptor serine suggest that both sites are engaged simultaneously to the cyclin and the Cdk, respectively, and eliminate a "bind and release" mechanism to increase the local concentration of the substrate. PS100, a peptide containing a Cy motif, acts as a competitive inhibitor of cyclin-Cdk complexes with a 15-fold lower K(i) for cyclin E-Cdk2 than for cyclin A-Cdk2. These results provide kinetic proof that a Cy motif located a minimal distance from the SPXK is essential for optimal phosphorylation by Cdks and suggest that small chemicals that mimic the Cy motif would be specific inhibitors of substrate recognition by cyclin-dependent kinases. PMID- 11067845 TI - The adapter type protein CMS/CD2AP binds to the proto-oncogenic protein c-Cbl through a tyrosine phosphorylation-regulated Src homology 3 domain interaction. AB - CMS/CD2AP is a cytoplasmic protein critical for the integrity of the kidney glomerular filtration and the T cell function. CMS contains domains and motifs characteristic for protein-protein interactions, and it is involved in the regulation of the actin cytoskeleton. We report here that the individual SH3 domains of CMS bind to phosphotyrosine proteins of approximately 80, 90, and 180 kDa in cell lysates stimulated with epidermal growth factor. The second SH3 domain of CMS bound specifically to a tyrosine-phosphorylated protein of 120 kDa, which we identified as the proto-oncoprotein c-Cbl. The c-Cbl-binding site for CMS mapped to the carboxyl terminus of c-Cbl and is different from the proline rich region known to bind SH3-containing proteins. CMS binding to c-Cbl was markedly attenuated in a tyrosine phosphorylation-defective c-Cbl mutant indicating that this interaction is dependent on the tyrosine phosphorylation of CMS. It also implies that CMS interacts with c-Cbl in an inducible fashion upon stimulation of a variety of cell-surface receptors. Immunofluorescence analysis revealed that both proteins colocalize at lamellipodia and leading edges of cells, and we propose that the interaction of CMS with c-Cbl offers a mechanism by which c-Cbl associates and regulates the actin cytoskeleton. PMID- 11067846 TI - A single nucleotide polymorphic mutation in the human mu-opioid receptor severely impairs receptor signaling. AB - Large scale sequencing of the human mu-opioid receptor (hMOR) gene has revealed polymorphic mutations that occur within the coding region. We have investigated whether the mutations N40D in the extracellular N-terminal region, N152D in the third transmembrane domain, and R265H and S268P in the third intracellular loop alter functional properties of the receptor expressed in mammalian cells. The N152D receptor was produced at low densities. Binding affinities of structurally diverse opioids (morphine, diprenorphine, DAMGO and CTOP) and the main endogenous opioid peptides (beta-endorphin, [Met]enkephalin, and dynorphin A) were not markedly changed in mutant receptors (<3-fold). Receptor signaling was strongly impaired in the S268P mutant, with a reduction of efficacy and potency of several agonists (DAMGO, beta-endorphin, and morphine) in two distinct functional assays. Signaling at N40D and R265H mutants was highly similar to wild type, and none of the mutations induced detectable constitutive activity. DAMGO-induced down regulation of receptor-binding sites, following 20 h of treatment, was identical in wild-type and mutant receptors. Our data show that natural sequence variations in hMOR gene have little influence on ligand binding or receptor down-regulation but could otherwise modify receptor density and signaling. Importantly, the S268P mutation represents a loss-of-function mutation for the human mu-opioid receptor, which may have an incidence on opioid-regulated behaviors or drug addiction in vivo. PMID- 11067847 TI - Characterization of a prostate-specific tyrosine phosphatase by mutagenesis and expression in human prostate cancer cells. AB - The cellular form of human prostatic acid phosphatase (PAcP) is a neutral protein tyrosine phosphatase (PTP) and may play a key role in regulating the growth and androgen responsiveness of prostate cancer cells. The functional role of the enzyme is at least due in part to its dephosphorylation of c-ErbB-2, an in vivo substrate of the enzyme. In this study, we investigated the molecular mechanism of phosphotyrosine dephosphorylation by cellular PAcP. We mutated several amino acid residues including one cysteine residue that was proposed to be involved in the PTP activity of the enzyme by serving as the phosphate acceptor. The cDNA constructs of mutant enzymes were transiently transfected into C-81 LNCaP and PC 3 human prostate cancer cells that lack the endogenous PAcP expression. The phosphotyrosine level of ErbB-2 in these transfected cells was subsequently analyzed. Our results demonstrated that the phosphotyrosine level of ErbB-2 in cells expressing H12A or D258A mutant PAcP is similar to that in control cells without PAcP expression, suggesting that these mutants are incapable of dephosphorylating ErbB-2. In contrast, cells expressing C183A, C281A, or wild type PAcP had a decreased phosphotyrosine level of ErbB-2, compared with the control cells. Similar results were obtained from in vitro dephosphorylation of immunoprecipitated ErbB-2 by these mutant enzymes. Furthermore, transient expression of C183A, C281A, or the wild-type enzyme, but not H12A or D258A, decreased the growth rate of C-81 LNCaP cells. The data collectively indicate that His-12 and Asp-258, but not Cys-183 or Cys-281, are required for the PTP activity of PAcP. PMID- 11067848 TI - Cloning, expression, and up-regulation of inducible rat prostaglandin e synthase during lipopolysaccharide-induced pyresis and adjuvant-induced arthritis. AB - We have cloned and expressed the inducible form of prostaglandin (PG) E synthase from rat and characterized its regulation of expression in several tissues after in vivo lipopoylsaccharide (LPS) challenge. The rat PGE synthase is 80% identical to the human enzyme at the amino acid level and catalyzes the conversion of PGH(2) to PGE(2) when overexpressed in Chinese hamster ovary K1 (CHO-K1) cells. PGE synthase activity was measured using [(3)H]PGH(2) as substrate and stannous chloride to terminate the reaction and convert all unreacted unstable PGH(2) to PGF(2alpha) before high pressure liquid chromatography analysis. We assessed the induction of PGE synthase in tissues from Harlan Sprague-Dawley rats after LPS induced pyresis in vivo. Rat PGE synthase was up-regulated at the mRNA level in lung, colon, brain, heart, testis, spleen, and seminal vesicles. Cyclooxygenase (COX)-2 and interleukin 1beta were also up-regulated in these tissues, although to different extents than PGE synthase. PGE synthase and COX-2 were also up regulated to the greatest extent in a rat model of adjuvant-induced arthritis. The RNA induction of PGE synthase in lung and the adjuvant-treated paw correlated with a 3.8- and 16-fold induction of protein seen in these tissues by immunoblot analysis. Because PGE synthase is a member of the membrane-associated proteins in eicosanoid and glutathione metabolism (MAPEG) family, of which leukotriene (LT) C(4) synthase and 5-lipoxygenase-activating protein are also members, we tested the effect of LTC(4) and the 5-lipoxygenase-activating protein inhibitor MK-886 on PGE synthase activity. LTC(4) and MK-886 were found to inhibit the activity with IC(50) values of 1.2 and 3.2 microm, respectively. The results demonstrate that PGE synthase is up-regulated in vivo after LPS or adjuvant administration and suggest that this is a key enzyme involved in the formation of PGE(2) in COX 2-mediated inflammatory and pyretic responses. PMID- 11067849 TI - A remarkably stable phosphorylated form of Ca2+-ATPase prepared from Ca2+-loaded and fluorescein isothiocyanate-labeled sarcoplasmic reticulum vesicles. AB - After the nucleotide binding domain in sarcoplasmic reticulum Ca2+-ATPase has been derivatized with fluorescein isothiocyanate at Lys-515, ATPase phosphorylation in the presence of a calcium gradient, with Ca2+ on the lumenal side but without Ca2+ on the cytosolic side, results in the formation of a species that exhibits exceptionally low probe fluorescence (Pick, U. (1981) FEBS Lett. 123, 131-136). We show here that, as long as the free calcium concentration on the cytosolic side is kept in the nanomolar range, this low fluorescence species is remarkably stable, even when the calcium gradient is subsequently dissipated by ionophore. This species is a Ca2+-free phosphorylated species. The kinetics of Ca2+ binding to it indicates that its transport sites are exposed to the cytosolic side of the membrane and retain a high affinity for Ca2+. Thus, in the ATPase catalytic cycle, an intrinsically transient phosphorylated species with transport sites occupied but not yet occluded must also have been stabilized by fluorescein isothiocyanate (FITC), possibly mimicking ADP. The low fluorescence mainly results from a change in FITC absorption. The Ca2+-free low fluorescence FITC-ATPase species remains stable after addition of thapsigargin in the absence or presence of decavanadate, or after solubilization with dodecylmaltoside. The remarkable stability of this phosphoenzyme species and the changes in FITC spectroscopic properties are discussed in terms of a putative FITC-mediated link between the nucleotide binding domain and the phosphorylation domain in Ca2+-ATPase, and the possible formation of a transition state-like conformation with a compact cytosolic head. These findings might open a path toward structural characterization of a stable phosphorylated form of Ca2+-ATPase for the first time, and thus to further insights into the pump's mechanism. PMID- 11067850 TI - Kinetics of CO and NO ligation with the Cys(331)-->Ala mutant of neuronal nitric oxide synthase. AB - Nitric-oxide synthases (NOS) catalyze the conversion of l-arginine to NO, which then stimulates many physiological processes. In the active form, each NOS is a dimer; each strand has both a heme-binding oxygenase domain and a reductase domain. In neuronal NOS (nNOS), there is a conserved cysteine motif (CX(4)C) that participates in a ZnS(4) center, which stabilizes the dimer interface and/or the flavoprotein-heme domain interface. Previously, the Cys(331) --> Ala mutant was produced, and it proved to be inactive in catalysis and to have structural defects that disrupt the binding of l-Arg and tetrahydrobiopterin (BH(4)). Because binding l-Arg and BH(4) to wild type nNOS profoundly affects CO binding with little effect on NO binding, ligand binding to the mutant was characterized as follows. 1) The mutant initially has behavior different from native protein but reminiscent of isolated heme domain subchains. 2) Adding l-Arg and BH(4) has little effect immediately but substantial effect after extended incubation. 3) Incubation for 12 h restores behavior similar but not quite identical to that of wild type nNOS. Such incubation was shown previously to restore most but not all catalytic activity. These kinetic studies substantiate the hypothesis that zinc content is related to a structural rather than a catalytic role in maintaining active nNOS. PMID- 11067851 TI - C-mannosylation and O-fucosylation of the thrombospondin type 1 module. AB - Thrombospondin-1 (TSP-1) is a multidomain protein that has been implicated in cell adhesion, motility, and growth. Some of these functions have been localized to the three thrombospondin type 1 repeats (TSRs), modules of approximately 60 amino acids in length with conserved Cys and Trp residues. The Trp residues occur in WXXW patterns, which are the recognition motifs for protein C-mannosylation. This modification involves the attachment of an alpha-mannosyl residue to the C-2 atom of the first tryptophan. Analysis of human platelet TSP-1 revealed that Trp 368, -420, -423, and -480 are C-mannosylated. Mannosylation also occurred in recombinant, baculovirally expressed TSR modules from Sf9 and "High Five" cells, contradictory to earlier reports that such cells do not carry out this reaction. In the course of these studies it was appreciated that the TSRs in TSP-1 undergo a second form of unusual glycosylation. By using a novel mass spectrometric approach, it was found that Ser-377, Thr-432, and Thr-489 in the motif CSX(S/T)CG carry the O-linked disaccharide Glc-Fuc-O-Ser/Thr. This is the first protein in which such a disaccharide has been identified, although protein O-fucosylation is well described in epidermal growth factor-like modules. Both C- and O glycosylations take place on residues that have been implicated in the interaction of TSP-1 with glycosaminoglycans or other cellular receptors. PMID- 11067852 TI - Identification, characterization, and functional analysis of heart-specific myosin light chain phosphatase small subunit. AB - Myosin light chain phosphatase consists of three subunits, a 38-kDa catalytic subunit, a large 110-130-kDa myosin binding subunit, and a small subunit of 20-21 kDa. The catalytic subunit and the large subunit have been well characterized. The small subunit has been cloned and studied from smooth muscle, but little is known about its function and specificity in the other muscles such as cardiac muscle. In this study, cDNAs for heart-specific small subunit isoforms, hHS M(21), were isolated and characterized. Evidence was obtained from an analysis of genome to suggest that the small subunit was the product of the same gene as the large subunit. Using permeabilized renal artery preparation and permeabilized cardiac myocytes, it was shown that the small subunit increased sensitivity to Ca(2+) in muscle contraction. It was also shown using an overlay assay that hHS M(21) bound the large subunit. Mapping experiments demonstrated that the binding domain and the domain involved in the increasing Ca(2+) sensitivity mapped to the same N-terminal region of hHS-M(21). These observations suggest that the heart specific small subunit hHS-M(21) plays a regulatory role in cardiac muscle contraction by its binding to the large subunit. PMID- 11067853 TI - Elevated triglyceride content diminishes the capacity of high density lipoprotein to deliver cholesteryl esters via the scavenger receptor class B type I (SR-BI). AB - The selective uptake of high density lipoprotein (HDL) cholesteryl ester (CE) by the scavenger receptor class B type I (SR-BI) is well documented. However, the effect of altered HDL composition, such as occurs in hyperlipidemia, on this important process is not known. This study investigated the impact of variable CE and triglyceride (TG) content on selective uptake. CE selective uptake by Y1 and HepG2 cells was strongly affected by modification of either the CE or TG content of HDL. Importantly, TG, like CE, was selectively taken up by a dose-dependent, saturable process in these cells. As shown by ACTH up-regulation and receptor overexpression experiments, SR-BI mediated the selective uptake of both CE and TG. With in vitro modified HDLs of varying CE and TG composition, the selective uptake of CE and TG was dependent on the abundance of each lipid within the HDL particle. Furthermore, total selective uptake (CE + TG) remained constant, indicating that these lipids competed for cellular uptake. These data support a novel mechanism whereby SR-BI binds HDL and mediates the incorporation of a nonspecific portion of the HDL lipid core. In this way, TG directly affects the ability of HDL to donate CE to cells. Processes that raise the TG/CE ratio of HDL will impair the delivery of CE to cells via this receptor and may compromise the efficiency of sterol balancing pathways such as reverse cholesterol transport. PMID- 11067854 TI - Angiotensin I-converting enzyme transition state stabilization by HIS1089: evidence for a catalytic mechanism distinct from other gluzincin metalloproteinases. AB - Angiotensin (Ang) I-converting enzyme (ACE) is a member of the gluzincin family of zinc metalloproteinases that contains two homologous catalytic domains. Both the N- and C-terminal domains are peptidyl-dipeptidases that catalyze Ang II formation and bradykinin degradation. Multiple sequence alignment was used to predict His(1089) as the catalytic residue in human ACE C-domain that, by analogy with the prototypical gluzincin, thermolysin, stabilizes the scissile carbonyl bond through a hydrogen bond during transition state binding. Site-directed mutagenesis was used to change His(1089) to Ala or Leu. At pH 7.5, with Ang I as substrate, k(cat)/K(m) values for these Ala and Leu mutants were 430 and 4,000 fold lower, respectively, compared with wild-type enzyme and were mainly due to a decrease in catalytic rate (k(cat)) with minor effects on ground state substrate binding (K(m)). A 120,000-fold decrease in the binding of lisinopril, a proposed transition state mimic, was also observed with the His(1089) --> Ala mutation. ACE C-domain-dependent cleavage of AcAFAA showed a pH optimum of 8.2. H1089A has a pH optimum of 5.5 with no pH dependence of its catalytic activity in the range 6.5-10.5, indicating that the His(1089) side chain allows ACE to function as an alkaline peptidyl-dipeptidase. Since transition state mutants of other gluzincins show pH optima shifts toward the alkaline, this effect of His(1089) on the ACE pH optimum and its ability to influence transition state binding of the sulfhydryl inhibitor captopril indicate that the catalytic mechanism of ACE is distinct from that of other gluzincins. PMID- 11067855 TI - Identification of a conserved motif in the yeast golgi GDP-mannose transporter required for binding to nucleotide sugar. AB - Glycoproteins and lipids in the Golgi complex are modified by the addition of sugars. In the yeast Saccharomyces cerevisiae, these terminal Golgi carbohydrate modifications primarily involve mannose additions that utilize GDP-mannose as the substrate. The transport of GDP-mannose from its site of synthesis in the cytosol into the lumen of the Golgi is mediated by the VRG4 gene product, a nucleotide sugar transporter that is a member of a large family of related membrane proteins. Loss of VRG4 function leads to lethality, but several viable vrg4 mutants were isolated whose GDP-mannose transport activity was reduced but not obliterated. Mutations in these alleles mapped to a region of the Vrg4 protein that is highly conserved among other GDP-mannose transporters but not other types of nucleotide sugar transporters. Here, we present evidence that suggest an involvement of this region of the protein in binding GDP-mannose. Most of the mutations that were introduced within this conserved domain, spanning amino acids 280-291 of Vrg4p, lead to lethality, and none interfere with Vrg4 protein stability, localization, or dimer formation. The null phenotype of these mutant vrg4 alleles can be complemented by their overexpression. Vesicles prepared from vrg4 mutant strains were reduced in luminal GDP-mannose transport activity, but this effect could be suppressed by increasing the concentration of GDP-mannose in vitro. Thus, either an increased substrate concentration, in vitro, or an increased Vrg4 protein concentration, in vivo, can suppress these vrg4 mutant phenotypes. Vrg4 proteins with alterations in this region were reduced in binding to guanosine 5'-[gamma-(32)P]triphosphate gamma-azidoanilide, a photoaffinity substrate analogue whose binding to Vrg4-HAp was specifically inhibited by GDP mannose. Taken together, these data are consistent with the model that amino acids in this region of the yeast GDP-mannose transporter mediate the recognition of or binding to nucleotide sugar prior to its transport into the Golgi. PMID- 11067856 TI - Formation of conjugated delta8,delta10-double bonds by delta12-oleic-acid desaturase-related enzymes: biosynthetic origin of calendic acid. AB - Divergent forms of the plant Delta(12)-oleic-acid desaturase (FAD2) have previously been shown to catalyze the formation of acetylenic bonds, epoxy groups, and conjugated Delta(11),Delta(13)-double bonds by modification of an existing Delta(12)-double bond in C(18) fatty acids. Here, we report a class of FAD2-related enzymes that modifies a Delta(9)-double bond to produce the conjugated trans-Delta(8),trans-Delta(10)-double bonds found in calendic acid (18:3Delta(8trans,10trans,12cis)), the major component of the seed oil of Calendula officinalis. Using an expressed sequence tag approach, cDNAs for two closely related FAD2-like enzymes, designated CoFADX-1 and CoFADX-2, were identified from a C. officinalis developing seed cDNA library. The deduced amino acid sequences of these polypeptides share 40-50% identity with those of other FAD2 and FAD2-related enzymes. Expression of either CoFADX-1 or CoFADX-2 in somatic soybean embryos resulted in the production of calendic acid. In embryos expressing CoFADX-2, calendic acid accumulated to as high as 22% (w/w) of the total fatty acids. In addition, expression of CoFADX-1 and CoFADX-2 in Saccharomyces cerevisiae was accompanied by calendic acid accumulation when induced cells were supplied exogenous linoleic acid (18:2Delta(9cis,12cis)). These results are thus consistent with a route of calendic acid synthesis involving modification of the Delta(9)-double bond of linoleic acid. Regiospecificity for Delta(9)-double bonds is unprecedented among FAD2-related enzymes and further expands the functional diversity found in this family of enzymes. PMID- 11067857 TI - The C terminus of annexin II mediates binding to F-actin. AB - Annexin II heterotetramer (AIIt) is a multifunctional Ca(2+)-binding protein composed of two 11-kDa subunits and two annexin II subunits. The annexin II subunit contains the binding sites for anionic phospholipids, heparin, and F actin, whereas the p11 subunit provides a regulatory function. The F-actin binding site is presently unknown. In the present study we have utilized site directed mutagenesis to create annexin II mutants with truncations in the C terminus of the molecule. Interestingly, a mutant annexin II lacking its C terminal 16, 13, or 9 amino acids was unable to bind to F-actin but still retained its ability to interact with both anionic phospholipids and heparin. Recombinant AIIt, composed of wild-type p11 subunits and the mutant annexin II subunits, was also unable to bundle F-actin. This loss of F-actin bundling activity was directly attributable to the inability of mutant AIIt to bind F actin. These results establish for the first time that the annexin II C-terminal amino acid residues, LLYLCGGDD, participate in F-actin binding. PMID- 11067858 TI - RNA-transfected dendritic cells in cancer immunotherapy. PMID- 11067859 TI - Therapeutic applications of transcription factor decoy oligonucleotides.